PMID- 16689353 TI - [Androgen and reproductive physiology]. PMID- 16689354 TI - [Testicular dysfunction]. PMID- 16689355 TI - [Androgens and androgen receptor in prostate cancer]. PMID- 16689356 TI - [Androgen insensitivity syndrome]. PMID- 16689357 TI - [LOH (late-onset hypogonadism]. PMID- 16689358 TI - [CRT. ICD. LVAS]. PMID- 16689359 TI - [Recent advances in treatment and the future problems to be solved]. AB - The goal of the treatment of chronic heart failure is improvement of patient's prognosis and QOL. To achieve this goal, new pharmacological and nonpharmacological approaches have been developed. ACE inhibitor - and beta blocker-treatment added to the standard therapy has been established to improve the prognosis, and ICD, cardiac resynchronization treatment (CRT) with biventricular pacing and LV assist device also contribute to the advances in the therapeutic progress. However, recent clinical trials for the new drugs which antagonize the endothelin receptors and TNF receptors failed to demonstrate the benefit of these new drugs. Establishment of the therapeutic strategies for treatment of diastolic heart failure is also another problem to be solved. PMID- 16689360 TI - [Molecular mechanisms of congestive heart failure]. AB - Although heart failure is a final common consequence of various heart diseases and is a leading cause of mortality worldwide, the precise molecules and signaling pathways that mediate heart failure progression are largely undefined. In this review, we discuss about the potential mechanisms of heart failure focusing on three subjects, (i) myocardial ischemia, (ii) cardiac muscle cell death, and (iii) abnormalities in calcium handling. These factors are not mutually independent but considered to contribute to the pathogenesis of contractile dysfunction and heart failure in a cooperative manner. Elucidation of molecular mechanisms of heart failure will lead to the development of novel therapeutic strategies for heart diseases. PMID- 16689361 TI - [Role of sympathoadrenal activations in congestive heart failure]. AB - Although sympathoneuronal activation plays a major role in maintaining circulatory homeostasis in decompensated heart failure, protracted activation adversely affects survival in patients with chronic heart failure. Endstage heart failure is associated with beta-adrenergic receptor subsensitivity phenomenon including downregulation of beta1-adrenergic receptors, uncoupling induced by receptor kinase activations, increased level of inhibitory guanine nucleotide binding protein. Abnormal calcium handling occurs as a result of dissociation of ryanodine receptors and its modulatory protein, FK506 binding proteins in addition to defective calcium uptake. Phosphoinositide 3-kinase gamma as well as defective neuronal uptake mechanism of norepinephrine have a pivotal role in mediating these alterations in adrenergic receptor signalings. In this regards, modulation of excessive adrenergic activations is essential to achieve successful management of patients with congestive heart failure. PMID- 16689362 TI - [Cardiac aldosterone]. AB - In heart failure, the heart carries out compensation of its functional deterioration using various hormonal actions. Many hormones are recently found to be synthesized and secreted from the failing or hypertensive heart. We have reported that the steroid synthesis is present in the heart; aldosterone but not dehydroepiandrosterone (DHEA) is secreted from the failing heart and contrarily DHEA but not aldosterone is secreted from the normal heart. Furthermore, we reported that adrenocorticotropic hormone (ACTH) is also secreted from the failing or hypertensive heart, suggesting that the hormones composing the hypothalamus-pituitary-adrenal(HPA) axis as well as the RAA system would be present in the heart. PMID- 16689363 TI - [Pathophysiological role of cytokines in heart failure]. AB - Plasma and myocardial levels of proinflammatory cytokines such as tumor necrosis factor(TNF)-alpha, interleukin 1beta, and interleukin 6 are elevated in patients with heart failure. Not only viral infection but also cardiac overload, ischemia, and neuro-humoral factors stimulate systemic and myocardial production of these cytokines. Administration of proinflammatory cytokines directly depresses myocardial contractility in vitro and in vivo. In addition, TNF and interleukin induce myocardial apoptosis and promote cardiac hypertrophy and fibrosis, suggesting that these cytokines are involved in the progression of cardiac remodeling. Vasoactive peptides such as endothelin and adrenomedullin also have cytokine-like functions in the heart, acting as autocrine and paracrine factors. Thus, proinflammatory cytokines and these peptides play important roles in the pathogenesis and pathophysiology of heart failure. PMID- 16689364 TI - [Oxidative stress and heart failure]. AB - The excessive reactive oxygen species (ROS), produced by the electron leak in mitochondria of failing myocardium, play an important role in development and progression of heart failure (HF) and cardiac remodeling, which is associated with induction of myocyte hypertrophy and apoptosis and activation of matrix metalloproteinases. Furthermore, ROS can damage mitochondrial DNA(mtDNA), and thus lead to mitochondrial dysfunction and additional ROS generation. Recently, we found that the overexpression of mitochondrial transcription factor A (TFAM), which is essential for mtDNA transcription and replication, ameliorates cardiac remodeling and failure. Therefore, the regulation of mitochondrial oxidative stress or manipulation of TFAM protein may provide a novel therapeutic strategy of HF. PMID- 16689365 TI - [Cardiac remodeling in chronic heart failure]. AB - Left ventricular remodeling plays an important role in the progression of chronic heart failure. The early remodeling is an adaptive response to the increased load(i.e. hypertension) and/or the loss of contractile component (i. e. myocardial infarction) in order to maintain the pumping capacity. This process includes cardiac hypertrophy associated with myocyte hypertrophy and myocardial fibrosis. However, the persistence of the inadequate overload results in the decompensation of the adaptive mechanism. Finally, left ventricle is enlarged and the systolic function is impaired. PMID- 16689366 TI - [Plasma B-type natriuretic peptide (BNP) measuring]. AB - Brain or B-type natriuretic peptide (BNP) is a potent natriuretic, diuretic and vasorelaxant peptide and inhibits sympathetic tone, the renin-angiotensin aldosterone system, and synthesis of vasoconstrictive molecules. The major source of plasma BNP is the cardiac ventricles. Elevated plasma BNP concentrations correlate with increased left ventricular (LV) filling pressure. Therefore BNP is a useful biomarkers as a screening tool for LV dysfunction. It also is a strong diagnostic indicator for both systolic and diastolic LV dysfunction. Measurement of plasma BNP is proved to be not only an efficient but also a cost effective screening tool for identifying patients with acute dyspnea of unknown etiology. PMID- 16689367 TI - [Echocardiography]. AB - Advancement of echocardiography has been remarkable recently and has been useful to diagnose and manage chronic heart failure. Tissue Doppler echocardiography can measure myocardial motion velocity thereby enabling us to assess global and regional myocardial function. Mitral annular velocity in early diastole has been reported to correlate with left ventricular diastolic relaxation. The ratio of pulsed Doppler mitral flow velocity to the mitral annular velocity in early diastole is used to assess pulmonary capillary wedge pressure or left ventricular end-diastolic pressure. Comparison of time to peak of myocardial systolic velocity among various segments provides us important information on ventricular dyssynchrony, which is crucial for the success of cardiac resynchronization therapy. Finally, real-time three-dimensional echocardiography has emerged as a new modality to assess cardiac anatomical abnormalities. It is also promising to measure ventricular volume accurately. PMID- 16689368 TI - [Cardiac imaging]. AB - Congestive heart failure is a state of inadequate cardiac function under various etiologies. Cardiac imaging plays an important role for accurate detection of heart failure, assessment of severity of LV function, and precise analysis of tissue function in vivo. Recently, multislice CT(MSCT), magnetic resonance imaging (MRI), and positron emission tomography(PET) have been rapidly developed for clinical use for assessing patients with congestive heart failure. MSCT has been used for accurate assessment of LV function. Due to high spatial resolution, MSCT permits assessment of coronary stenosis without cardiac catheterization. MRI permits assessment of LV function and also tissue function. Particularly, infarcted myocardium is accurately delineated as an area of delayed enhancement by contrast enhancement MRI study. PET has been used for accurate assessment of myocardial viability based on the persistence of myocardial glucose metabolism. In addition, a various new PET tracers permit molecular and cellular function, such as neurotransmission and receptor function in vivo. These new imaging technique has a potential role for assessing risk stratification and providing appropriate treatment strategy. PMID- 16689369 TI - [Invasive diagnostic technique and technology for heart failure]. AB - Technology and techniques of cardiac catheterization to assess the ventricular function have been developed in the recent decades. The approach for assessing function is based on the measurement of cardiac output which depends on preload, afterload and ventricular contractility. Clinical application of Frank-Starling principle is useful for characterizing human heart failure. Conventionally, the isovolumic phase index(peak positive dP/dt) and ejection phase index (ejection fraction) have been applied to evaluate ventricular systolic function. Recently, ventricular diastolic function as well as systolic function can be estimated using ventricular pressure-volume loops obtained by simultaneous measurement of pressure and volume. The slope of end-systolic pressure-volume relation represents a load-independent index of ventricular contractility and assessing ventricular end-diastolic pressure-volume relation can express ventricular passive diastolic compliance for the evaluation of human heart failure. PMID- 16689370 TI - [Recent advance of exercise test in clinical cardiology concerning heart failure]. AB - Present review focused on the recent advance and clinical application of exercise test in cardiology. Exercise intolerance is considered as one of clinical essential feature in patients with heart failure. Cardiac dysfunction induces systemic disorders including motor muscle and vasculature, and then limits exercise performance through direct and indirect pathway. Parameters of exercise test consist of subjective indexes such as NYHA functional class, and objective evaluations such as cardiopulmonary exercise test(CPX). Especially, we discussed the parameters of CPX provided many indexes, for example peak VO2, VE/VCO2 slope and deltaVO2/Work rate. These various indexes have different clinical meanings among each other. Low value of peak VO2, which reflected both disturbances of central and peripheral factors, is reported to result in poor survival rate. Although VE/VCO2 slope similarly predicts the prognosis, it thought to be mediated through a different way. We concluded that exercise test is important to understand the pathophysiological feature of heart failure as a systemic disease. PMID- 16689371 TI - [Pharmacological therapeutics based on molecular mechanisms]. AB - Pharmacological treatment of chronic heart failure has considerably changed according as a shift of the pathophysiological concept. During the past two decades, it became clear that neurohumoral stimulations play an important role in the progression of heart failure by promoting cardiac remodeling. Administration of angiotensin-converting enzyme-inhibitors, angiotensin II receptor blockers, and beta blockers is now an essential treatment that reduces morbidity and mortality. In spite of an increase in available treatments, the prognosis of symptomatic heart failure remains to be poor. To establish newer and successful treatments, further advancement in our understanding of the molecular mechanisms underlying the pathogenesis of heart failure will be required. Currently, several novel approaches are being tested targeting prevention of cardiac remodeling and myocardial dysfunction. Cardiomyocyte protection by hematopoietic cytokines and stem cell-based therapy may have a promising therapeutic potential. PMID- 16689372 TI - [ACE inhibitor]. AB - Chronic heart failure (CHF) is a clinical complex showing end -stage of several cardiovascular disorders. Patients with CHF are disclosed to both of high mortality and high morbidity. Most of them repeat frequently the hospitalization. In CHF patients, neurohumoral factors such as renin -angiotensin -aldosterone systems (RAAS) are enhanced, and they result in high level of angiotensin, aldosterone and catecholamine in the cardiovascular tissue. Most of the hormones are known as cardiotoxic agent. The activated RAAS is closely linked with progression of cardiac remodeling. Thus, ACE inhibitor can block this linkage. Recently, several randomized controlled trials in large scale reveal that the ACE inhibitor is a beneficial tool for not only CHF therapy but also CHF prevention. The blocking effects by ACE inhibitor are playing a crucial role in releasing the CHF patients from several burdens. PMID- 16689373 TI - [Use of angiotensin type II receptor blockers as a potential strategy for chronic heart failure]. AB - Angiotensin II is one of the most important key molecules for cardiac remodeling and apoptosis, culminating in heart failure. Therefore, angiotensin II receptor blockers (ARBs) have been expected to improve the mortality and morbidity in patients with heart failure more effectively strict elimination of the effects of angiotensin II compared with angiotensin-converting-enzyme inhibitors (ACEI). Here, we summarize the current clinical outcomes obtained from large scale trials using ARBs as a strategy of the treatment of heart failure, and discuss the pharmacological differences between ARB and ACEI and the future direction of the treatment of heart failure. PMID- 16689374 TI - [Aldosterone antagonist]. PMID- 16689375 TI - [Future perspectives of beta-blockers in chronic heart failure]. AB - The efficacy of beta-blockers for improving the survival rate of patients with chronic heart failure has been confirmed by results deriving from a series of randomized controlled trials (RCTs) such as MERIT-HF, CIBIS-II, and COPERNICUS. Thus, treatment guidelines recommend that beta-blockers should be used in all patients with heart failure resulting from left ventricular systolic dysfunction who can tolerate with beta-blockers. Despite these guidelines, actual rates of beta-blocker prescription has been lower than expected, and when used, beta blockers are generally prescribed in doses lower than what has been shown to reduce morbidity and mortality in RCTs. In the MUCHA trial conducted in Japan, the beta-blocker carvedilol decreased the cardiovascular risk for hospitalization in a dose-dependent manner. The 5 mg/day dose achieved a remarkable reduction that was nearly as great as the 20 mg/day dose. Whether this low-dose therapy was sufficient was not fully investigated by the MUCHA trial. Moreover, there is an ethnic difference in the effective dose between patients in Japan and Western countries, which may depend on a difference in beta1-receptor sensitivity. J-CHF trial may suggest that low doses of carvedilol under 2.5 mg/day have a beneficial effect. This trial is a randomized multi-center parallel group dose-comparison study, and with sub-studies such as an analysis of genetic polymorphisms on voluntary participation. This trial may serve as a tailored approach for Japanese physicians in the clinical management of heart failure. PMID- 16689376 TI - [Inotropic agents]. AB - We now know pharmacological negative inotropic agent, i. e. beta-blocker, increase cardiac contractility when given chronically to patients with chronic heart failure. In contrast, positive inotropic agents have consistently been shown to increase mortality when given chronically. The routine use of inotropes in either acute or the chronic setting is not supported by the available data. However, some appropriate uses of inotropes are as temporary use of refractory acutely decompensated heart failure, or as a bridge to definitive treatment, or palliative measure in a subset of patients with end-stage heart failure. PMID- 16689377 TI - [Diuretics]. AB - Diuretics interfere with the sodium retention of heart failure by inhibiting the reabsorption of sodium in the renal tubes, increase urine output and decrease physical signs of fluid retention in patients with heart failure (HF). Diuretics are the only drugs used for the treatment of HF that can adequately control the fluid retention of HF. Loop diuretics, potassium-sparing agents and thiazides are used for treatment. Tolvaptan, vasopressin receptor antagonists, is a new drug and long-term clinical trials are under way to determine the role. The use of inappropriate doses of diuretics will lead to result in fluid retention, renal insufficiency with ACE inhibitors and ARBs, increase the risk of treatment with beta blockers or volume contraction, increase the risk of hypotention and diminish the response to ACE inhibitors and ARBs. Appropriate use of diuretics is important in the success of the treatment of HF. PMID- 16689378 TI - [Cardiac resynchronization therapy]. AB - Despite recent advances in pharmacologic treatment for heart failure, the prognosis of patients with chronic heart failure remains poor. One third of patients with chronic heart failure have intraventricular conduction delay. The ventricular mechanical dyssynchrony based on intraventricular conduction delay worsen ventricular systolic dysfunction. Cardiac resynchronization therapy(CRT) through biventricular pacing significantly improves symptoms, exercise tolerance, hemodynamics, hospitalization for heart failure and mortality. However, it is estimated that 30 % of patients do not respond to CRT. A direct assessment of mechanical dyssynchrony with echocardiography seems more important than QRS duration in selecting appropriate patients for CRT. PMID- 16689379 TI - [Implantable cardioverter-defibrillator in patients with chronic heart failure]. AB - Sudden cardiac death (SCD) is a major cause of death in patients with chronic heart failure. The implantable cardioverter-defibrillator (ICD) effectively treats malignant ventricular tachyarrhythmias and reduces significantly the total mortality as well as the incidence of SCD in heart failure patients. It is evident that ICD is indicated for the secondary prevention of SCD. There is growing evidence for the use of the ICD for the primary prevention of SCD in patients with LV systolic dysfunction without documented arrhythmia. However, the efficacy of ICD seems to be modest in patients with advanced heart failure. Individualized combined therapies such as ICD plus amiodarone and ICD plus cardiac resynchronization therapy are necessary for advanced heart failure patients. It is doubtful whether ICD is indicated for MADIT II and SCD -HeFT population in Japan, where the incidence of SCD is thought to be lower than the Western countries. PMID- 16689380 TI - [Ventricular assist system]. AB - Despite the recent remarkable advances in the medical treatment for severe end stage heart failure, we still need a more powerful therapy to save patients presenting with cardiogenic shock or progressive cardiac deterioration. The implantation of ventricular assist system (VAS) is one of the available strategies in such patients and it has been used as a bridge to recovery or a bridge to transplantation. Because of the severe shortage of brain -dead donors, the role of VAS is now expanding into an alternative to transplantation, which is called a destination therapy. This review summarizes the current application and the future direction of VAS therapy for end -stage heart failure in Japan. PMID- 16689381 TI - [Exercise therapy for heart failure]. AB - In chronic heart failure of CAD, therapeutic approach will be available either with drugs or exercise. With exercise, coronary risk factors such as BP, lipid, DM and obesity will be controlled. In addition, ischemia will also be controlled by decreasing oxygen demand related to BP and HR, and with increasing oxygen supply by increased ECNOS gene expression, collateral formation and regression of coronary stenosis. Infarct size is also reported to be decreased by increasing MnSOD in the cell by exercise. Prognosis of CHF is also good in various evidence of exercise therapy. Recent advances of molecular biology have revealed various mechanisms of exercise effect. Thus, exercise if properly prescribed without provoking ischemia will be basically and clinically effective therapy for patients with CHF. PMID- 16689382 TI - [Home oxygen therapy (HOT) for patients with heart failure associated with sleep disturbed breathing]. AB - Recently, there is increasing evidence that sleep apnea may adversely affect pathophysiology and outcomes of congestive heart failure (CHF). Repetitive nocturnal apneas may worsen CHF through a number of mechanisms including the repetitive arterial oxygen desaturation, increased left ventricular afterload, or an activation of sympathetic nervous system. Although central sleep apnea (CSA) is relatively rare, prospective studies revealed that 33 to 82 % of patients with CHF have evidence of CSA and characteristic Cheyne-Stokes respiration (CSR). We assessed an efficacy of nasal O2 therapy at night using a conventional O2 concentrator in ambulatory patients with stable CHF and CSR. O2 resulted in a significant improvement of sleep together with an increase in left ventricular function and quality of life. Therefore, home oxygen therapy(HOT) can be a valuable nonpharmacological option for the treatment of patients with CHF and CSR CSA. PMID- 16689383 TI - [Surgical ventricular restoration]. AB - Surgical left ventricular restoration has been introduced to reshape the dilated cardiac chamber and improve cardiac function for patients with severe congestive heart failure secondary to ischemic or idiopathic dilated cardiomyopathy. The operations for ischemic cardiomyopathy (Dor, SAVER, etc) provide significant improvements of cardiac function and good short- and long-term survival. Ongoing prospective study (STICH trial) may make those more standard procedures. On the contrary, the operation for idiopathic dilated cardiomyopathy (Batista procedure) carries high risks of operative mortality and recurrence of congestive heart failure. Modifications of operative indications and surgical techniques are under investigation. PMID- 16689384 TI - [Characteristics of the chronic heart failure in the elderly]. AB - Chronic heart failure (CHF) in the elderly is a significant healthcare problem. Nearly half of CHF in this age group is diastolic heart failure with preserved left ventricular ejection fraction. However there is the limitation of the evidence-based therapy for this important group of patients. In this article, the treatments for the diastolic heart failure recommended in the ACC/AHA guidelines for the CHF (1998, 2001, 2005 versions) are described and common problems regarding the diagnosis and management of CHF in the elderly is also discussed. Furthermore, the recent topics about the treatment of diastolic heart failure are presented. PMID- 16689385 TI - [Treatment of life-threatening arrhythmias in patients with chronic heart failure]. AB - Several proven medical therapies such as beta-blockers and angiotensin converting enzyme inhibitors play important roles in preventing sudden death of patients with heart failure. Therapy with amiodarone and therapy with implantable cardioverter defibrillator (ICD) have developed specifically to prevent sudden death by life-threatening arrhythmias. But recently many large-scale clinical trials have proved that ICD therapy improves prognosis of patients with heart failure better than amiodarone therapy. Because ICD is not a treatment to prevent the occurrence of ventricular arrhythmias, it may not necessarily improve quality of life of patients. In that respect catheter ablation may play a role in preventing life-threatening arrhythmias. PMID- 16689386 TI - [Challenges towards regenerative medicine]. AB - Regenerative medicine is a promising approach to treat patients with severe cardiac failure. Since embryonic stem cells (ES cells) easily differentiate into cardiomyocytes, ES cells are thought to be a good candidate resource for cardiac cell transplantation therapy. However, molecular mechanism of cardiac differentiation is still largely unknown. Here we discuss our present approach to understand the mechanism of cardiogenesis at the molecular level as well as novel genes and cascades that are important for cardiac differentiation. Further observation will help to establish the new strategy of regenerative medicine for patients with cardiac failure. PMID- 16689387 TI - [Molecular mechanism of diabetic nephropathy]. AB - Diabetic nephropathy(DN) is the most common cause of end-stage renal disease, and is characterized by excessive accumulation of extracellular matrix in the kidney. Chronic hyperglycemia is the main cause of DN. Multiple theories have been proposed to explain the adverse effect of hyperglycemia, including an increased flux through the polyol pathway, excessive formation of advanced glycation end products, oxidative stress, and activation of the protein kinase C (PKC) pathway. In this article, we review evidence that supports these theories, and focus our attention on the PKC theory. Therapeutic interventions to inhibit PKC are effective in reducing DN in diabetic animals. Currently, clinical trials are in progress to determine the efficacy of a PKC beta inhibitor for the treatment of diabetic complications including DN. PKC beta inhibitors may be included as a first line treatment for DN in a near future. PMID- 16689388 TI - [Spontaneous cerebrospinal fluid hypovolemia]. PMID- 16689389 TI - [Surgical techniques for neuroendoscopic procedures]. PMID- 16689390 TI - [Fusion imaging of 3D MR cisternography/angiography for differential diagnosis of internal carotid-posterior communicating artery aneurysms and infundibular dilations]. AB - By using a fusion imaging of three-dimensional (3D) magnetic resonance cisternography (MRC) and coregistered magnetic resonance angiography (MRA), protrusions at the bifurcation of the internal carotid-posterior communication artery, detected by MRA, were investigated to differentiate between the infundibular dilations and aneurysms. The MRA, obtained by the 3D time-of-flight sequence, showed the flow-related arterial structures by means of an inflow effect mainly induced by the peak systolic flow velocity. The MRC, obtained by 3D fast spin-echo sequence, depicted the contours of the vascular structures within the cisternal space in contrast to the surrounding cerebrospinal fluid. Fusion images of 3D MRC/MRA showed the anatomy of protrusions from the different viewpoints with flow-related intraluminal images (MRA) in conjunction with outer wall configuration images of the vascular structures (MRC). This imaging technique may be useful to differentiate the infundibular dilations from the internal carotid-posterior communicating artery aneurysms. PMID- 16689391 TI - [The correlation between lacunes and microbleeds on magnetic resonance imaging in consecutive 180 patients]. AB - BACKGROUND AND OBJECTIVE: Microbleeds on T2*-weighted magnetic resonance imaging (MRI) represent a hemorrhagic type of small vessel disease. Small vessel disease causes both intracerebral hemorrhages and lacunar infarctions. We studied clinical background and MRI findings of the patients to clarify the correlation between microbleeds and lacunes. METHOD: This study consisted of 180 consecutive patients who underwent brain MRI using 1.5T system in our hospital for a year. We obtained T2*-weighted gradient-echo imaging as well as T1 and T2-weighted images. We statistically identified the factors related to the presence of microbleeds in all patients. The distribution of lacunes and microbleeds on MRI was compared to clarify the correlation of the lesions in the patients who had both lesions. RESULT: The overall prevalence of microbleeds was 41.1% (74 of 180 patients). Logistic regression analysis indicated that previous stroke, leukoaraiosis and lacunes were significantly correlated with microbleeds. In the patients who have both microbleeds and lacunes, lesions are tended to locate in thalamus and basal ganglia, especially incidence of lacunes are significantly greater compared with other regions. Thirty-six of 398 lesions (9.05%) diagnosed as lacunes with T1- and T2-weighted imaging were demonstrated as microbleeds with T2*-weighted gradient-echo imaging. CONCLUSION: Lacunes, leukoaraiosis as a hypertensive change on MRI had statistically significant correlation with the presence of microbleeds. T2*-weighted gradient-echo imaging should be included in the imaging protocol for cerebrovascular disease, because T1- and T2-weighted imaging recognizing some of the microbleeds as lacunar infarction. PMID- 16689392 TI - [Traumatic panhypopituitarism: case report]. AB - We describe a case of traumatic panhypopituitarism following head injury. Generally considered, posttraumatic hypopituitarism occurs in patients who have suffered from severe head injury. However there were a few case reports of panhypopituitarism due to mild and moderate head injury. A 51-year-old male presented with a history of blunt head injury caused by a concrete block hitting his head directly during work. On admission, initial Glasgow Coma Scale was 14. Open depressed skull fracture was suspected. Emergency craniectomy and debridement were performed. Ten days after surgery, hypothermia, lethargy and appetite loss were manifested. Endocrinological examination showed panhypopituitarism with diabetes insipidus. MRI revealed ruptured pituitary stalk and pituitary gland hemorrhage. Coronal and sagittal MRI was helpful for the diagnosis of traumatic panhypopituitarism. General condition was recovered by hormone replacement therapy. It is important for medical staff carefully to observe vital signs and clinical symptoms, even if mild brain injury. Pituitary function test should also be undergone, if panhypopituitarism was suspected from clinical condition. PMID- 16689393 TI - [Stab injury of the brain by kitchen knife]. AB - A 32-year-old male attempted suicide by stabbing his forehead with a kitchen knife, and was sent to our hospital. On admission, he was confused (20-30/JCS), but was able to move his both extremities. Skull X-p, CT scan indicated that the kitchen knife penetrated the frontal bone, reached the cerebrum by way of the frontal sinus. Cerebral angiography was performed, and showed no vascular lesions. Emergent craniotomy was performed and the kitchen knife was removed with minimum movement. Traumatic intracerebral hematoma was removed, and injured frontal sinus, dura mater and frontal bone were repaired. Postoperative course was excellent and he was discharged with the wounds well healded, no neurological deficits on the 17th day after the operation. There are several reported cases of such stab injury. Stab injury of the brain was discussed in the literature. PMID- 16689394 TI - [Stent-assisted coiling of an acutely ruptured large aneurysm of the internal carotid artery: case report]. AB - A 48-year-old female was referred to our hospital for the management of a ruptured 22-mm-diameter internal carotid aneurysm, located immediately distal to the ophthalmic artery. The right internal carotid artery was completely involved in the aneurysm. Because the right carotid siphon was relatively nontortuous stent-assisted coiling of the aneurysm was attempted 4 days after subarachnoid hemorrhage. A new coil-type coronary stent, a DRIVER stent, was deployed after microcatheter insertion into the aneurysm dome under systemic heparinization, followed by dome packing using Guglielmi detachable coils. Systemic heparinization was continued for 18 hours postoperatively, and aspirin (100 mg/day) was administered orally immediately after the embolization. However, 4 hours after heparin administration cessation, left hemiparesis became apparent. Immediate magnetic resonance angiography revealed a right internal carotid artery occlusion. Diffusion-weighted images demonstrated ischemic spots in the watershed zones of the right cerebral hemisphere. Following an immediate thrombolysis of the right internal carotid artery, systemic heparinization was continued for 5 days, and aspirin and ticlopidine were administrated orally. The patient was discharged on day 37, following the recovery from left hemiparesis within ten days. A DRIVER stent is a low profile coil-type coronary stent, which can be used for the stent-assisted coiling of large internal carotid aneurysms, located distal to the carotid siphon. It seems necessary to continue systemic heparinization for more than 2 days and to administer increased dose or two types of antiplatelet after the stent-assisted coiling of large cerebral aneurysms. PMID- 16689395 TI - [Gliomatosis cerebri. Report of two cases]. AB - Gliomatosis cerebri is a rare tumor of the central nervous system, and here we report two cases of this tumor. Case 1: A 41-year-old female was admitted to our department for evaluation of activity loss and mental changes. MR imaging revealed a high intensity symmetrical lesion in the bilateral frontal and temporal regions, thalamus and midbrain on FLAIR images. The patient had no neurological deficits and no abnormal findings in clinical laboratory data, including data for the cerebral spinal fluid. A specimen obtained by open biopsy revealed widespread infiltration of neuronal structures by small astrocytic cells, although without destruction of the neuronal structures. The patient was diagnosed with gliomatosis cerebri, and surgery and whole brain radiation at 44Gy were performed. The MRI lesion showed some shrinkage 20 months after surgery, and the KPS score was 90, the same as that before admission. Case 2: A 56-year-old male was admitted for numbness of the face and extremities. MR imaging revealed a high intensity lesion in the bilateral thalamus and a ringed enhanced lesion in the right thalamus. A specimen obtained by open biopsy revealed anaplastic oligodendroglioma, which was diagnosed as gliomatosis cerebri. Radiation at 54Gy, chemotherapy (ACNU, vincristine) and gamma-knife surgery were performed, and two months later MR imaging showed that the tumor (including the ringed enhanced lesion) had shrunk markedly. His KPS was 90 at 13 months after onset. These cases suggest that radiation therapy is effective for gliomatosis cerebri. PMID- 16689396 TI - [Cavernous hemangioma growing continuously from the spinal epidural space into the paravertebral muscles: case report]. AB - A 19-year-old male was admitted to our hospital with a 5 year history of right low back pain. MR images showed an epidural mass which located on the posterolateral side at the T12-L1 level and extended through the right intervertebral foramen into the paraspinal muscles. Excision of the intramuscular and epidural lesions was performed and his symptom quickly improved. Histopathological examination of the mass revealed a cavernous hemangioma. It may affect any part of neuraxis, however cavernous hemangiomas located at the spinal epidural space are rare and particularly the lesions which grow continuously into the paravertebral muscles are very rare. Radiological presentation of this case was confusing in the preoperative diagnosis. It is important that we know the clinical, developmental and radiological characteristics of cavernous hemangiomas. PMID- 16689397 TI - [Battle against reviewers, yet know thyself]. PMID- 16689399 TI - Family management style and the challenge of moving from conceptualization to measurement. AB - This article describes the process of translating the Family Management Style (FMS) Framework into a measure of FMS Survey. The conceptual underpinnings of the FMS Survey are briefly described as are the steps for translating the FMS major components and dimensions into a questionnaire to assess the family response to a childhood chronic illness. Content validity testing has been completed on the FMS Survey. Future plans include full-scale field testing of remaining psychometric properties. PMID- 16689400 TI - Family Management Style Framework: a new tool with potential to assess families who have children with brain tumors. AB - Qualitative studies of families with children who have cancer or other serious illnesses have found that families often come to view their child and their lives as normal. They manage illness-related demands using family management styles that sustain usual patterns of family and child functioning. Few studies have addressed the family management styles of families who express less satisfaction with family and child functioning or who are identified by health care professionals as having difficulty with family functioning. Such families are likely to be overrepresented among those whose children are being treated for brain tumors that entail extremely burdensome treatments as well as a range of unfavorable prognoses and long-term sequelae. In fact, little is known about how these families manage on a day-to-day basis and how the interdisciplinary team can best provide supportive care to optimize their functioning. The purpose of this article is to present the Family Management Styles Framework as a tool that is useful in both clinical practice and research for assessing families who have children with cancer, including those with brain tumors. PMID- 16689401 TI - Clinical application of family management styles to families of children with cancer. AB - The potential clinical application of family management styles for working with families who have children with cancer is discussed. Case studies are used to illustrate the usefulness and clinical application of the model. PMID- 16689402 TI - Use of family management styles in family intervention research. AB - Family management styles (FMSs) explain some of the complexities embedded in a family with a child who has chronic illness. The FMS typologies provide descriptions of family adjustment and management of care. These 5 distinct patterns may be valuable in tailoring and evaluating family interventions in research. PMID- 16689403 TI - Consensus statements: the Family Management Style Framework and its use with families of children with cancer. PMID- 16689404 TI - Understanding the clinical implications of minimal residual disease in childhood leukemia. AB - Improved laboratory techniques now allow a more sensitive detection of leukemia cells at designated intervals throughout therapy. Using flow cytometry and polymerase chain reaction, it is possible to detect 1 leukemic cell among 10(4) normal cells (1 leukemia cell in 10,000 normal cells), representing a 100-fold greater sensitivity than morphological examination in acute lymphoblastic leukemia (ALL). Recently, it has been shown that the molecular presence of persistent acute lymphoblastic leukemia at the end of remission therapy is a poor indicator of clinical outcome. Now similar studies are being performed in acute myeloid leukemia (AML). While the sensitivity using flow cytometry is less in AML than in ALL (able to detect 1 leukemic cell among 1000 normal cells in AML), persistent or minimal residual AML provides the clinician guidance with future treatment recommendations. Minimal residual disease (MRD) is now considered an important indicator response of disease response to treatment. As such, MRD once considered a research variable is now influencing treatment decisions. Therefore, it is imperative that the nurse have an understanding of the newer techniques to study residual leukemia and their clinical implications for patients and their families. PMID- 16689405 TI - Can end of life care for the pediatric patient suffering with escalating and intractable symptoms be improved? AB - Over twelve thousand children are diagnosed each year with cancer, and approximately 2200 children die each year from the disease. A percentage of these patients experiences escalating and intractable distress with symptoms that include pain, dyspnea, and agitation. These symptoms may continue for hours to days. Intractable symptoms of pain, agitation, and dyspnea can be very distressing to the patient, family, and staff and often a challenge for the physicians and nursing staff to treat. To meet this challenge, The Dana-Farber Cancer Institute/Children's Hospital Cancer Care Program has made it a priority to create a process of care that includes identifying barriers to care and the development of an end-of-life (EOL) rapid response model that includes guidelines and physician-templated orders for rapid escalation of opioids. The goal of this quality-improvement initiative was to develop a model of care that would enable the caregivers to provide effective comfort care to any patient experiencing symptoms of rapid escalation of pain, dyspnea, and agitation. A model of care was created to overcome barriers to care. The model includes role clarification, "Guidelines for the Management of Escalating Pain/Dyspnea/Agitation at the End of Life," and "Rapid Titration-Templated Physician Orders." Staff feedback was solicited relative to the content, format, and usability of the guidelines and templated orders. The physician and nursing staff reported that they found the templated orders and guidelines very helpful and effective and suggested only a few edits. A retrospective chart review is currently under way. The purpose of this chart review is to systematically document and compare the record of management of rapidly escalating symptoms of pain and/or dyspnea and/or agitation prior to and after instituting the EOL Rapid Response Model of Care. Care of the EOL patient experiencing symptoms of pain, dyspnea, and agitation is challenging. The EOL Rapid Response Model of Care outlines a process of care and provides recommendations and templated physician orders for rapid titration of opioids. PMID- 16689406 TI - Building a leadership infrastructure: the next step in the evolution of hospital systems. PMID- 16689407 TI - Focus on physician satisfaction to improve quality, boost patient satisfaction, contribute to bottom line. PMID- 16689408 TI - When you are old...all alone? PMID- 16689409 TI - Lower limb lymphedema: experiences and perceptions of cancer patients in the late palliative stage. AB - Lower limb lymphedema (LLL) is a common but neglected problem in palliative cancer patients. No studies have focused on these patients' experiences of lymphedema. The aims of this study were to explore patients' experiences regarding LLL and how they manage to deal with this in the late palliative stage. METHODS: Thirteen patients with cancer-related LLL were included to satisfy a maximum variation sampling strategy. Interviews were analyzed using a qualitative phenomenographic method. RESULTS: LLL influenced the patients' thoughts about the future. Body image was often strongly influenced. Interactions with other persons were perceived as both positive and negative, and a range of coping strategies were expressed. CONCLUSION: LLL can exert a considerable influence on the physical experiences and the psychosocial situation of cancer patients in palliative care. Areas in need of increased education, attention, and further research are highlighted. PMID- 16689410 TI - Indicators of poor quality end-of-life cancer care in Ontario. AB - This study measures the proportion of cancer patients in Ontario, Canada, with intensive care unit (ICU) admissions, emergency room (ER) visits, or chemotherapy in the last two weeks of life. We used the Ontario Cancer Registry to identify a cohort of cancer patients who died in 2001. These cases were then linked to administrative sources of data to measure each indicator, and to describe the associated clinical and health service factors. In the cohort, 27% had at least one ER visit and 5% had an ICU visit in the last two weeks of life. Of those who received chemotherapy in the last six months, 16% received chemotherapy in the last two weeks of life. Receiving a home care visit in the last six months of life, or a physician house call or a palliative care assessment in the last two weeks of life was consistently associated with decreased odds of each of the indicators. Our results indicate that a significant proportion of Ontario cancer patients have indicators of poor quality end-of-life care. Certain health care factors may influence these indicators. PMID- 16689411 TI - Quality of care for residents dying in Ontario long-term care facilities: findings from a survey of directors of care. AB - The purpose of this study was to collect information on the practice of end-of life (EOL) care in long-term care (LTC) facilities in the Province of Ontario, Canada. A cross-sectional survey of directors of care in all licensed LTC facilities in the province was conducted between September 2003 and April 2004. Directors of care from 426 (76% response rate) facilities completed the postal survey questionnaire. The survey results identified communication problems between service providers and families, inadequate staffing levels to provide quality care to dying residents, and the need for training to improve staff skills in providing EOL care. Directors of care endorsed the use of a number of strategies that would improve the care of dying residents. Logistic regression analysis identified the eight most important items predictive of facility staff having the ability to provide quality EOL care. The findings contribute to the current discussion on policies for meeting the care needs of residents in LTC facilities until life's end. PMID- 16689412 TI - Being a family in the midst of living and dying. AB - The aim of this study was to illuminate the way of being a family when one family member is in the midst of living and dying. A family systems frame and a life world perspective were used in interviews with five families. A qualitative analysis inspired by Giorgi revealed dialectic and dynamic processes in constant motion within and between the continua being in affinity-being in loneliness, being in power-being in helplessness, and being in continuity-being in disruption. When families were moving in the direction of being in affinity, power, and continuity, these seemed to be prerequisites for enduring their challenged life situation and for giving them a kind of repose. When the movements were in the opposite direction, existential and emotional suffering were manifested as individual embodied experiences such as depression and anxiety. PMID- 16689413 TI - The last word: family members' descriptions of end-of-life care in long-term care facilities. AB - A postal survey was used to collect data from family members of deceased residents of six long-term care (LTC) facilities in order to explore end-of-life (EOL) care using the Family Perception of Care Scale. This article reports on the results of thematic analysis of family member comments provided while completing the survey. Family comments fell into two themes: (1) appreciation for care and (2) concerns with care. The appreciation for care theme included the following subthemes: psychosocial support, family care, and spiritual care. The concerns with care theme included the subthemes: physical care, staffing levels, staff knowledge, physician availability, communication, and physical environment. This study identified the need for improvement in EOL care skills among LTC staff and attending physicians. As such, there is a need to implement continuing education to address these issues. PMID- 16689414 TI - Validation of the Korean version of the McMaster Quality of Life Scale in terminal cancer patients. AB - The purpose of this study was to develop and validate a Korean version of McMaster Quality of Life Scale (K-MQLS) suitable for evaluating clinical hospice and palliative care. The McMaster Quality of Life Scale (MQLS) is a brief and comprehensive scale that is used to assess cancer patients receiving palliative care due to poor physical condition. To further develop the K-MQLS, we followed rigorous international translation steps and performed validity, reliability, and sensitivity analyses. The results of our study show that the K-MQLS is an efficient tool in terms of its validity, reliability, and sensitivity for the measurement of the health-related quality of life during the palliative phase. This developed tool could be used in research or clinical settings to assess health-related quality of life in Korean palliative care patients. PMID- 16689415 TI - Improving access to hospice and palliative care for patients near the end of life: present status and future direction. PMID- 16689416 TI - A possible model of mentoring the establishment of a home palliative care unit in a "resource-strapped" country. PMID- 16689417 TI - Can fentanyl be systemically absorbed when administered vaginally? A feasibility study. PMID- 16689418 TI - Win or lose? Percutaneous nephrostomy for a terminal-stage cervical-cancer patient featuring obstructive uropathy. PMID- 16689419 TI - Christiana Hospital honored for groundbreaking project. PMID- 16689421 TI - Small hospitals take on big projects. PMID- 16689420 TI - Making the case for electronic records. PMID- 16689422 TI - I.T. modernizes house calls. PMID- 16689423 TI - Provider searches for accurate data. PMID- 16689424 TI - Readers' perspectives. Are National health information network and regional health information networks destined to fail unless a national patient identifer is mandated? PMID- 16689425 TI - Minn., Calif. say no to exec perks. States question pay for UnitedHealth's CEO, directors. PMID- 16689426 TI - Stating the case for coverage. Frustrated with federal efforts to solve the uninsured problem, many states are taking it upon themselves to find solutions. AB - States are leading the search for solutions to the uninsured problem, explorng various paths to universal coverage. In Massachusetts, which recently passed a near-universal plan, it took months to lay the groundwork, and comments were sought from a wide variety of groups. "You can't just look at the most recent accomplishment without understanding what brought us to this point," says Michael Miller, left. PMID- 16689427 TI - AHA wants more time. PPS quality-reporting deadline called too tight. PMID- 16689428 TI - Adding another role. JCAHO's Brown says Beryl position is clear of conflict. PMID- 16689429 TI - CEO turnover slows--for now. As boomers begin retiring the tide may shift again. PMID- 16689430 TI - Expanding in Ohio. WellPoint set to enter state's Medicaid HMO market. PMID- 16689431 TI - Minn.'s Hatch blasts Blues deal. Says expensive disease-management deal not working. PMID- 16689432 TI - Alvarado prosecutor's dilemma. Lawyers say third time's not usually a charm. PMID- 16689433 TI - Staffing deal brings risks. Jackson buys rival mired in bankruptcy, lawsuits. PMID- 16689434 TI - Not-so-benign neglect. Federal inaction leaves states to experiment on covering more uninsured. PMID- 16689435 TI - A history lesson not learned. Four factors stop us from covering the uninsured, but the pressure is mounting. PMID- 16689436 TI - High-octane hospitals. Solucient's annual list of performance-improvement leaders shows organizationwide attention key to financial and operational gains. PMID- 16689437 TI - By the numbers. Top business graduate schools for physician-executives. Data collected in March 2006 for the 2005-06 school year. PMID- 16689438 TI - Sex education attitudes and outcomes among North American women. AB - Attitudes and outcomes of sex education received by North American women are examined via an Internet survey (N = 1,400). Mean age was 19.5, with 24% reporting one or more unplanned pregnancies. Women were more satisfied with sex education from informal sources than from parents, schools, and physicians. Those receiving sex education from parents or schools reported fewer pregnancies and abortions. In school, women receiving a combination of contraceptive and abstinence education and those receiving primarily abstinence education were least likely to experience unplanned pregnancy. Religious identification was significantly related to unplanned pregnancy and type of sex education received from parents. These factors seem to play a significant role in reducing unplanned pregnancy and abortion. PMID- 16689439 TI - Consumption of sexual dialogue and content on television and adolescent sexual outcomes: multiethnic findings. AB - The purposes of this study were to examine group differences in sexual media consumption and to explore how well adolescents' sexual attitudes and behavior can be explained by viewing of sexually suggestive dialogue (SD) and explicit sexual content (SC) in television media. Participants were 473 male and female high school adolescents who were racially, economically, and age diverse. Television viewing was coded for SD and SC. Both length of time spent watching SD and SC and explicitness of the SD and SC to which adolescents were exposed were factored into the coding scheme. In this sample, SD and SC consumption varied by subgroup and explained variance in some sexual attitudes and behavior variables but not others, and in some demographic subgroups but not others. PMID- 16689440 TI - Gender differences in victimized homeless adolescents. AB - Most of what we know about sexual abuse comes from efforts to examine female children victimized by men. Although some researchers have identified similarities between male and female victims of sexual abuse, few studies have examined gender-specific factors associated with sexual health practices among homeless adolescents. The aim of this study was to explore how gender and history of sexual abuse influence cognitive-perceptual and behavioral factors associated with sexual health practices of homeless adolescents. This study was a secondary analysis of data collected for a cross-sectional study of the sexual health practices of homeless adolescents. The sample consisted of 414 youths (104 males who reported sexual abuse and 124 who did not; and 95 females who reported sexual abuse and 75 who did not; 16 did not provide these data). Homeless adolescent females with a history of sexual abuse scored higher (indicating a shorter perspective) on a measure of future time perspective than females with no sexual abuse. Males who reported no sexual abuse scored higher than abused females on perceived health status and higher than abused males on assertive communication. With respect to perceived health status, males who reported no sexual abuse scored significantly higher than females who reported sexual abuse (p = .04). Males with no sexual abuse had significantly higher assertive communication scores than did males who had experienced sexual abuse (p = .015). We found that male and female abuse victims differ in terms of their cognitive-perceptual and behavioral factors associated with sexual health practices. Early identification of those who have been abused is critical so that interventions can be developed. Effective short-term interventions are needed for the adolescent victims of Child Sexual Abuse (CSA), particularly those who are homeless and prone to further sexual victimization. PMID- 16689441 TI - Understanding Girls' Circle as an intervention on perceived social support, body image, self-efficacy, locus of control, and self-esteem. AB - The Girls' Circle is a support group for adolescent girls developed by Beth Hossfeld and Giovanna Taormina as a unique program that addresses the needs of girls by focusing on increasing connections, building empathic skills, and developing resiliency. The present study evaluates the effectiveness of the Girls' Circle intervention on improving social support, body image, locus of control, self-efficacy, and self-esteem. Sixty-three girls from 9 support groups (comprising 5 to 15 girls each) across the United States completed the Multidimensional Scale of Perceived Social Support, the Body Parts Satisfaction Scale, the Nowicki-Strickland Personal Reaction Survey, Schwarzer's General Self Efficacy Scale, and the Rosenberg Self-Esteem scale both before and after the 10 week Girls' Circle program. Results revealed a significant increase in social support, body image, and self-efficacy after completion of the program. PMID- 16689442 TI - Adolescent girls' perceived barriers to participation in physical activity. AB - Research shows a decline in physical activity levels during adolescence, particularly among girls. This study explored perceived barriers to participation in moderate and vigorous physical activity among adolescent girls who live in a large ethnoracially and socioeconomically diverse city. A total of 73 adolescent girls in Toronto participated in 1 of 7 focus group sessions which were audiotaped and transcribed verbatim, and qualitative software was used to facilitate data analysis. A constant comparison approach was used to inductively develop themes. Participants' perceived barriers to participating in physical activity included: lack of time; involvement in technology-related activities; influence peers, parents and teachers; concern about safety; inaccessibility of facilities and cost of using them; competition; and body-centered issues. The results suggest that strategies to address specific intrapersonal, social environmental, and physical-environmental barriers to physical activity participation should be used in physical activity promotion programs for adolescent girls. PMID- 16689443 TI - African American child-women: nutrition theory revisited. AB - Past research indicates a significantly higher prevalence of early sexual maturation in African American (AA) girls, which is associated with a number of psychological and behavioral problems as well as with health problems such as childhood obesity and diabetes. Both nutrition and body image perceptions have never before been empirically investigated in the context of early puberty. The present study analyzed nutritional differences among AA girls who experience early sexual maturation with those who do not, as well as the dynamics of early sexual maturation, food consumption, and body image perceptions of AA girls. Participants were 45 AA girls ages 7-10 years, and guardians who were recruited from Boys and Girls Clubs in the southeastern part of the U.S. The Continuing Survey of Food Intakes by Individuals and Fallon and Rozin's (1985) Body Image Checklist was used to assess the food intake and body image perceptions of the young girls. Evidence of early sexual maturation was operationalized as the parental report of development of any secondary sexual characteristics (breasts, hips, pubic hair) in the young girl. A two-day food intake recall was conducted over a representative weekday period. Results revealed a significant difference between the calcium and fiber consumption of the AA girls who experienced early sexual maturation versus those who did not. Also, results indicated a relationship between parental characteristics, household shopping practices, and food consumption of the young girls. These findings are important and can be used to predict, prepare, and educate the AA population as well as establish appropriate support systems for these child-women. PMID- 16689444 TI - Evaluating "baby think it over" infant simulators: a comparison group study. AB - To test the efficacy of Baby-Think-It-Over (BTIO) infant simulators, two versions of a sexuality education program were compared. While the program was designed to include BTIO as an important teaching technique, two schools (49 students) opted not to use them. These students completed all elements of the program except the BTIO activities. Their results were compared to students from three similar, neighboring schools that completed the regular program (including BTIO). Knowledge about sex, attitudes toward teen sex, parent-adolescent communication, and sexual behavior were measured on self-report instruments at the beginning and end of the three-week program. No significant differences were found between the two groups on any of the four measures on the pretest or posttest. This study and similar recent reports call into question the educational value of expensive infant simulators. PMID- 16689445 TI - The effect of perceived parental involvement and the use of growth-fostering relationships on self-concept in adolescents participating in gear up. AB - Adolescence is marked by physical, cognitive, and social changes which affect how individuals perceive themselves. However, individuals do integrate their perceptions with those of significant others. Adolescents' self-concept is influenced by their relationships with family, peers, mentors, and community groups. In this study, the characteristics of these relationships are examined as well as the perception of parental involvement. The effect of perceived parental involvement and the use of growth-fostering relationships on self-concept was examined using 78 adolescents who were participating in GEAR UP (Gaining Early Awareness of Readiness for Undergraduate Programs). Results support the influence of family, peers, mentors, and involvement in community groups on adolescent self concept. PMID- 16689446 TI - Adult helping qualities preferred by adolescents. AB - Most studies of therapeutic alliance have focused on adult populations and been written from the therapist's perspective. Thus, there is a clear need for studies of therapeutic alliance that focus on adolescent populations from the perspective of the adolescents. The current study is an exploratory process investigation using a focus group methodology with a nonclinical sample of adolescents to determine which traits in adults might foster alliance, with the assumption that the same traits would apply to therapeutic settings. A content analytic approach was used and 12 adult qualities found to be preferred by adolescents from most to least cited were: respect, time shared, openness, role characteristics, recognition, guidance, identification, trust, freedom, like/ dislike, responsibility, and familiarity. PMID- 16689447 TI - Ethnic identity and reconciliation: two main tasks for the young in Bosnia Herzegovina. AB - The saliency of ethnicity and ethnic identity is influenced by contextual circumstances. In Bosnia-Herzegovina, due to the current ethno-political situation, ethnicity and ethnic identity most likely are important aspects of adolescents' lives. The main purpose of this study is to describe a group of young Mostarians in relation to ethnic identity and reconciliation. Focus is on the participants' perspectives on these subjects and the meanings they attribute to the concepts. A total of 89 young Mostarians are included in the study. A third of the participants are members of a local peace- and reconciliation building organization, Koraci Nade. Results show that ethnic identity is highly salient for the group. While participants demonstrate strong emotional attachment to their ethnic group, exploration of ethnic identity is less reported. With regard to reconciliation, the participants perceive it as very desirable and show a personal and integrated relation to the concept. However, their definitions of the concept are vague as to what constitutes the process of reconciliation. Although of great importance to the members, participation in the peace-building organization was not found to enhance either ethnic identity or cross-ethnic friendships. PMID- 16689448 TI - Service-learning, personal development, and social commitment: a case study of university students in Hong Kong. AB - Service-learning, which combines academic study with community service, is becoming increasingly popular throughout the world. It is ideally suited to achieving both the personal and academic goals of students and the broader goals of civic responsibility and social justice. This paper describes the design of a local service-learning program the author implemented at a university in Hong Kong. Based on survey data collected from 93 university student participants in the program, it illustrates the impact of service-learning on student outcomes. Results show that the majority of the students benefitted as follows: (1) By developing personal autonomy through real world experiences, students develop a recognition of and faith in their potential. It enhances self-assurance, assumption of new responsibilities, and achievement of individual growth. (2) Students move toward becoming responsible citizens and agents of social change. By learning to care for deprived groups in the community, they are assuming meaningful roles and responding to real issues in ways that have a long-lasting impact on their own lives. Recommendations, based on the shortcomings we have witnessed and the changes we have implemented, are also made. PMID- 16689449 TI - The characteristics of separation-individuation in Turkish High School students. AB - The aim of the present study was examine the characteristics of separation individuation in Turkish high school students and to investigate the contribution of sociodemographic variables on this second individuation process of adolescence. The sample consisted of 618 adolescents between the ages of 14 and 18 in three urban and two rural high schools (338 females and 280 males). Measures used included a demographic questionnaire and the Separation Individuation Test of Adolescence (SITA), developed by Levin, Green, and Millon (1986). Results indicated that the 16-year-old group had significantly higher mean scores on the Engulfment Anxiety, Dependency Denial, and Rejection Expectancy subscales than the 15-year-old group. Males had significantly higher scores on the Practicing-Mirroring subscale than girls. Tenth graders had significantly higher mean scores on the Practicing-Mirroring, Nurturance Seeking, Peer Enmeshment, Teacher Enmeshment, and Healthy Separation subscales but the mean scores on the Dependency Denial and Engulfment Anxiety subscales decreased. The means scores on the Practicing-Mirroring, Dependency, Denial, Separation Anxiety, Teacher Enmeshment, and Rejection Expectancy subscales were significantly different among the socioeconomic status groups. Also, rural adolescents can be distinguished from urban counterparts by their increased tendency to perceive themselves as self-centered, to experience separation anxiety, to seek close interpersonal ties with caretakers, teachers, and peers, and by an integration of needs for dependence and independence. The general pattern of results investigating the separation-individuation development of Turkish adolescents suggested that compared with individualistic Western cultures, Turkish culture stressed the importance of connection as well as separation and psychic restructuring and interpersonal relatedness changes leading to an autonomous self within relational contexts. PMID- 16689450 TI - An examination of self-presentational concern of Turkish adolescents: an example of physical education setting. AB - The purpose of this study was to examine self-presentational concern of Turkish adolescents with regard to the gender composition of physical education (PE) class and also to investigate the attitude toward PE and PE class preferences of Turkish adolescents based on this concern. A total of 1,807 eighth-grade students (936 females and 871 males) completed the Attitudes Toward Physical Education Scale and Social Physique Anxiety Scale (SPAS). The findings indicated that although self-presentational concern did not differ according to the gender composition of PE class, there was a significant difference in class preference among adolescents based on this concern with most of the females who had a high self-presentational concern preferring noncoed PE class. Additionally, no significant difference was found in attitude toward PE among self-presentational concern groups. These findings are discussed in relation to the sex composition of PE class and self-presentational concerns of adolescents. PMID- 16689451 TI - [Molecular diagnosis/molecular therapy]. PMID- 16689452 TI - [The plasma cell myeloma--molecular pathogenesis and target therapies]. AB - The cells of the malignant clone of plasmacell myeloma have cytogenetic aberrations in a substantial number of cases. Many of these abnormal karyotypes are predictive for an unfavorable outcome. Gene mutations and abnormal gene expression, particularly of oncogenes and tumor suppressor genes, are often observed in myeloma cells. The cross talk between the myeloma cells and the bone marrow microenvironment plays an important role for growth and survival of the tumor cells. As a consequence of this cell-to-cell-interaction, several cytokines are secreted. The intracellular signaling, evoked by these cytokines, leads to continuous growth and proliferation and inhibition of apoptosis. Since these molecular pathways have been defined, many new targets for therapeutical interventions become obvious. Some molecules, directed against cytokines, are under early clinical investigation. Medicaments intervening in the cross talk between the myeloma cell and the bone marrow stroma as Thalidomide, Lenalidomide or Bortezomib are already available. Many of the myeloma patients suffer from bone disease. Some new drugs inhibiting the differentiation and activation of osteoclasts are evaluated in clinical trials. These molecules will be an important contribution against the painful bone disease of plasmacell myeloma. PMID- 16689453 TI - [Genetic diagnosis of acute leukemias--a practical guide]. AB - Acute leukemias are the consequence of accumulation of immature precursor cells. Genetic alterations are cause of uncontrolled cell proliferation. They can be detected by conventional cytogenetics and molecular methods at diagnosis. Classification of acute leukemias according to the World Health Organization depend on the knowledge of the presence of these aberrations. Furthemore they are important prognostic parameters influencing therapy intensity. Genetic abnormalities can be targets of novel therapeutic strategies. Detection and monitoring of genetic abnormalities in acute leukemias are of major importance for correct management of patients. PMID- 16689454 TI - [Molecular targets in colon cancer]. AB - Colorectal cancer is the second leading cause of cancer death in Switzerland. The nihilism that dominated the treatment of these patients for decades has been replaced by a measure of enthusiasm, given recent therapeutic advances. New anticancer drugs such as irinotecan and oxaliplatin have changed the standard chemotherapy treatment of metastatic colorectal cancer. However, the real hype has come from molecular targeted therapy. Identification of cellular processes characteristic of colon cancer has permitted therapeutic targeting with favorable therapeutic index. Inhibition of the epidermal growth factor receptor in the clinic has provided proof of principle that interruption of signal transduction cascades in patients has therapeutic potential. Angiogenesis, especially the vascular endothelial growth factor pathway, has been proven to be another highly successful molecular target. In this article, we will review molecular targets, which are under active clinical investigation in colon cancer. PMID- 16689455 TI - [Tyrosine kinase inhibitors for the treatment of CML]. AB - Chronic myelogenous leukemia is characterized by the Philadelphia-chromosome, a shortened chromosome 22 which is the result of a reciprocal translocation between chromosome 9 and 22. The fusion gene is called BCR-ABL. After transcription and translation the constitutively activated p210 BCR-ABL oncoprotein is formed. This leads to uncontrolled activation of the ABL tyrosin kinase. Deregulated cellular proliferation and diminished apoptosis of BCR-ABL transformed cells is the result. Expression of the BCR-ABL oncoprotein is sufficient and necessary for the development of a CML phenotype. Imatinib mesylate (Glivec) is a small molecule that binds to the ATP pocket of ABL and blocks downstream signalling events. Imatinib is very effective in the treatment of CML in all stages of the disease. Patients with newly diagnosed chronic phase CML were randomized to imatinib or to interferon plus cytarabine in the IRIS trial. Imatinib showed significantly superior tolerability, hematologic and cytogenetic resposes and increased time to progression. In patients with advanced phase CML, imatinib is less effective and response duration is short. Median overall survival of blast crisis patients is 6.9 months only. Additional BCR-ABL independent chromosomal abnormalities are common in advanced phase CML and result in resistance to imatinib. BCR-ABL kinase domaine mutations are frequently found in imatinib resistant patients and confer diminished sensitivity to imatinib. Second generation, more powerful ABL kinase inhibitors, which are effective against most of the known mutations are currently tested in clinical trials. PMID- 16689456 TI - [Therapeutic advances in breast cancer]. AB - The treatment of breast cancer has made significant improvements during the past ten years. For early breast cancer with a clinically negative axilla sentinel node biopsy has become the preferred approach. For endocrine therapy of postmenopausal patients the selective aromatase inhibitors have become standard in metastatic as well as in early breast cancer. Trastuzumab (Herceptin) plays an important role in the treatment of HER2-positive breast cancer in the metastatic and since 2005 also in the adjuvant setting. When chemotherapy is used to treat metastatic breast cancer drug combinations are superior to monotherapy only in terms of response rates. By contrast, in the adjuvant setting combination drug therapy is the standard. New methods of tissue analysis including expression patterns of mRNA and proteins are promising research strategies to further advance the field. PMID- 16689457 TI - [Cancer immunotherapy--current status]. AB - Cancer immunotherapy includes passive and active strategies. Passive immunotherapy such as the use of therapeutic monoclonal antibodies, and in a broader sense also of other immunological effector molecules, such as interferon alpha is clinically established. The efficacy of passive immunotherapy attests to the fact that the immune system can successfully fight cancer. The logical next step is therefore to develop strategies for active immunotherapy, i.e. "vaccines against cancer". This review focuses on the current status of active immunotherapy with respect to clinical application. Although active immunotherapy is still in the experimental stage, the data are highly encouraging and it is expected that vaccination will soon become part of cancer management. PMID- 16689458 TI - [Tissue banks for molecular diagnostics]. AB - The application of modern high throughput-technologies in genome and proteome research permits the identification of molecular, disease-associated mechanisms. Research data are mostly derived from experiments in which cell lines or animal models are used. In order to translate this knowledge into clinical usage it is necessary to examine numerous, well-documented tissues. Tissue collections that are appropriate for such investigations exist, particularly in pathology institutes. These tissues are routinely fixed informalin and embedded in paraffin and in some cases additional tissue pieces are also kept frozen for future diagnostic purposes. The linking of tissues with corresponding clinico pathological data and the use of the tissue microarray technology will enable rapid selection of new potential biomarkers for all kinds of diseases. By continuing the upgrading of tissue banks and by introducing quality controls, a significant potential for molecular diagnostics will be available in the future. This will contribute enormously to uncover and to establish novel therapeutical possibilities. PMID- 16689459 TI - [The diagnostic and predictive role of kit (CD117)]. AB - KIT (CD117) is a 145-KD transmembrane glycoprotein that is the product of the kit gene. As a member of the subclass III family of receptor tyrosine kinases, KIT is closely related to the receptors for platelet derived growth factor alpha and beta (PDGF-A and B), macrophage colony stimulating factor (M-CSF), and FLT3 ligand. The ligand for KIT, stem cell factor (SCF), also known as steel factor or mast cell growth factor promotes the dimerisation and autophosphorylation of KIT receptors. The phosphorylated tyrosine residues provide binding sites for signal molecules that contain SH2 domains. KIT mediated signal transduction is critical for the normal development and survival of haematopoietic progenitor cells, mast cells, interstitial cells of Cajal (intestinal pacemaker cells), melanocytes and germ cells. Upon differentiation, KIT expression is lost with the exception of mast cells, melanocytes and interstitial cells of Cajal. The detection of CD117 expression is of paramount diagnostic relevance in gastrointestinal stromal tumors (GIST). About 95% of all GISTs are immunohistochemically CD117 positive. The vast majority of all other sarcoma, carcinoma and also lymphoma are CD117 negative. Therefore, CD117 expression has a high sensitivity and specificity for the diagnosis of GIST. Moreover, activating mutations of KIT tyrosine kinase play a crucial pathogenetic role in GIST 80 to 85% of all GIST's contain activating mutations, primarily in Exon's 11 and 9 of the kit gene. Since the resulting mutant isoformes are sensitive to inhibition by imatinib (glivec), a specific tyrosine kinase inhibitor, the detection of a specific mutation has also a high predictive value. Besides GIST mastocytoses and seminomas are the neoplasias that most commonly express CD117. In contrast to GIST however, these two neoplasias contain mutations in different exons and are only partly imatinib sensitive. Moreover, CD117 expression is by no means entirely specific for these entities. It can also be detected in adenoid cystic carcinomas, thymic carcinomas and melanomas. Very rarely (< 5%) it can also be observed in other carcinomas and sarcomas. However, in the great majority of these cases with a CD117 protein expression there is no corresponding gene mutation of kit. Importantly, the lack of an activating mutation of KIT tyrosine kinase is good evidence that imatinib will not be effective. In other words, detection of sole CD117 protein expression is no solid basis for targeted therapy. The molecular pathological detection of CD117 expression in combination with the corresponding mutational status in patients with GIST (and other tumors) paradigmatically highlights the importance of modern molecular diagnostics in the era of targeted therapy. PMID- 16689460 TI - [Assessment of translocations in routine diagnostics of a surgical pathology unit]. AB - According to the current WHO classification of haematopoietic and lymphoid tissue, recurrent genetic aberrations constitute diagnostic markers for the histomorphological diagnosis of malignancies in these tissues. In lymphoma, translocations represent such genetic aberrations. Apart from their diagnostic impact, there is increasing evidence for their prognostic and predictive value. Despite the numerous translocations known in Non-Hodgkin's lymphoma (NHL), few are applied in routine diagnostics on a regular basis. To date, translocations have gained particular significance in B-cell NHLs, in contrast to T-cell NHLs where few are characterised. Refined tissue processing facilitates determination of translocations not only in fresh tissue, but increasingly also informalin fixed, paraffin embedded, and even decalcified biopsies. Hybridisation procedures (Southern blotting, FISH) and gene amplification methods (PCR, RT-PCR) are the primary molecular techniques employed to identify translocations. The article deals with the advantages and disadvantages of the currently used molecular techniques for the investigation of translocations in routine hematopathology diagnostics. PMID- 16689462 TI - Tune up your website. PMID- 16689461 TI - IV infusions. PMID- 16689463 TI - Hospitals are hiring again. PMID- 16689464 TI - Getting patients to commit. PMID- 16689465 TI - Is this pension plan right for you? PMID- 16689467 TI - What my mechanic taught me about being a doctor. PMID- 16689466 TI - Why EHRs falter. PMID- 16689468 TI - How to choose (and use) a collection agency. PMID- 16689469 TI - Adding ancilaries. Counseling services. PMID- 16689470 TI - Me? A substanee abuser. PMID- 16689471 TI - Documenting when you're on call. PMID- 16689472 TI - HIV/AIDS in Pakistan: has the explosion begun? PMID- 16689473 TI - Human immunodeficiency virus-associated disorders in Pakistan. AB - We have documented evidence of the human immunodeficiency virus (HIV) infection in four individuals including a blood donor and three members of a family (husband, wife, and a child) in Karachi, Pakistan. Our data indicate that HIV has been transmitted to the wife of the seropositive male, a drug abuser, in the recent past and that she has passed the virus to her newborn child perinatally/ transplacentally. The two seropositive males (blood donor and drug abuser) were diagnosed clinically as having persistent generalized lymphadenopathy, an AIDS related condition (ARC), and the child presented with developmental disorders and neurologic manifestations. This is the first report of AIDS/ARC cases from Pakistan that can be directly related to HIV infection and its heterosexual transmission. PMID- 16689474 TI - A view of HIV--infection in Karachi. AB - A prospective study on the prevalence of HIV-I infection in Karachi, Pakistan was conducted over a period of six years (1986-1992). Over 15,000 individual samples and more than 32,000 donor units of individuals residing in Karachi at the time of sample collection were tested for HIV-I infection by our screening test EIA which revealed a positivity rate of 0.23% and 0.003% in individual and donor units respectively by Western Blot. We divided patients into four groups A,B,C and D based on the most plausible cause of transmission. The largest number of positive patients belonged to group B, who were of either foreign origin or expatriates or Pakistanis settled abroad. They comprised approximately 67% of the total positive cases and were subjected to testing on strong clinical grounds. In individuals of other groups like group A and D, there was history of travel abroad from time to time. The only positive donor unit (group C) belonged to a person who had been living in Middle East for the last 10-12 years. The last group D comprised of samples that were directly sent to us without complete history, except for the fact that they had been traveling back and forth. The large majority of patients fell in 20-50 years age group. Despite the limitations of this study, we conclude that the prevalence of HIV is steadily increasing in our population and so far, we have not been able to find an indigenous case of AIDS in our series (JPMA 44:8, 1994). PMID- 16689475 TI - Understanding the HIV/AIDS context in Pakistan. AB - The social context of sexual relations is important in understanding the AIDS epidemic. So far, HIV in Pakistan has spread by heterosexual contact and blood transfusions. The magnitude of the problem is difficult to assess but health authorities estimate between 10,000 to 12,000 HIV infected people. This paper outlines that rapid urbanization, together with single migrant workers, deported HIV infected expatriates, exploitation of women and easy availability of narcotic drugs, especially in the metropolis of Karachi, are some important factors that may be responsible for the spread of HIV in Pakistan (Venereology 1995;8 [3]:160 3). PMID- 16689476 TI - HIV seroprevalence surveys in Pakistan. PMID- 16689477 TI - Seroprevalence of HIV, HBV and syphilis and associated risk behaviours in male transvestites (Hijras) in Karachi, Pakistan. AB - In Pakistan, male sex workers are predominantly transvestites and transsexuals known as Hijras. In 1998 in Karachi, Pakistan, we studied the seroprevalence of HIV, HBsAg and syphilis and associated risk factors in Hijras. Study subjects were verbally administered a structured questionnaire that determined risk behaviours for sexually transmitted and blood-borne diseases and knowledge of AIDS/STDs. After pre-test counselling, verbal consent was taken for serological testing. The results were provided on a one-on-one post-test counselling session. Three hundred male transvestites were approached; all agreed to answer the questionnaire, 208 consented to blood testing. Of 300, 81% acknowledged commercial sex with men. Of 208, prevalence of syphilis was 37%; HBsAg 3.4%; HIV 0%. The prevalence of HIV and hepatitis B virus (HBV) is low in transvestite sex workers but that of syphilis is high. Intervention programmes implemented at this stage can have an impact on HIV and STD prevention (Int J STD AIDS 1990;10:300 4). PMID- 16689478 TI - HIV-infected workers deported from the Gulf States: impact on Southern Pakistan. AB - HIV prevalence is still very low in Pakistan, but its south Asian location and subgroups with recognized lifestyle risk factors suggest that Pakistan will experience expanded diffusion of HIV. We report the frequency of HIV infections identified by the AIDS Control Programme in the Sindh province of Pakistan. Most HIV positive cases currently reported to the Sindh AIDS Control Programme are found among Pakistani workers deported from the Gulf States and among foreigners. The 58 returned workers with HIV represent 61 to 86% of reported cases in any given year during the 1996-1998 period. Five wives of returning workers have been identified with HIV. Expatriate workers in the Gulf States are tested for HIV routinely, unlike other subgroups in Pakistan. Considering the risk of HIV/AIDS due to regular introduction of HIV from returned workers, and the limited awareness surrounding sexual health and HIV/STD transmission issues in Pakistan, intervention programmes targeted at overseas workers need to be implemented to control the expansion of the HIV epidemic in Pakistan (Int J STD AIDS 1999;10:812 14). PMID- 16689479 TI - Seroprevalence of HBV, HCV and HIV infections among college going first time voluntary blood donors. AB - OBJECTIVE: College going first time voluntary blood donors were tested for HBV, HCV and HIV infections, to know the seroprevalence of these diseases among them. METHODS: Blood donation camps were organized at 18 colleges and universities of the city and a total of 612 samples from first time donors were tested for HbsAg, anti-HCV, anti-HIV 1, 2 antibodies on Abbott Diagnostic systems. RESULTS: Thirteen voluntary blood donors (2.21% with 95% confidence limit 1.2-3.52%) were HbsAg and 3 (0.5% with 95% confidence limit 0.12-1.33%) were anti HCV positive. None of the donors was positive for HIV infection. CONCLUSION: Seroprevalence of HBV, HCV and HIV infections among college going students is significantly low (<3.0%) than 30% seroprevalence among paid donors and 7% among family/replacement blood donors. There is a need to educate, motivate and recruit college going students to regularly donate blood to ensure relatively safe supply of blood for transfusion services (JPMA 50:269, 2000). PMID- 16689480 TI - Evaluation of blood bank practices in Karachi, Pakistan, and the government's response. AB - BACKGROUND: National legislation in Pakistan regulating blood banks has been introduced several times, but has never been passed. To support provincial-level efforts to develop legislation we conducted a study to evaluate blood-banking practices in Karachi, Pakistan, to identify areas that could be improved. METHODS: Thirty-seven blood banks were randomly selected from a list of 87 Karachi blood banks. The research team interviewed blood bank personnel, inspected available facilities and equipment, and observed blood collection using structured questionnaires and observation forms. RESULTS: Of the 37 selected facilities, 25 were operational and 24 agreed to participate. Twelve (50%) of the facilities reported regularly utilizing paid blood donors, while only six (25%) actively recruited volunteer donors. During observation only 8% of facilities asked donors about injecting drug use, and none asked donors any questions about high-risk sexual behaviour. While 95% of blood banks had appropriate equipment and reagents to screen for hepatitis B, only 55% could screen for HIV and 23% for hepatitis C. Twenty-nine percent of the facilities were storing blood products outside the WHO recommended temperature limits. IMPLICATIONS: Practives at most Karachi blood banks fell well below WHO standards. Findings from this study were instrumental in developing and passing legislation to regulate blood transfusion throughout Sindh Province, and suggest a method for improving blood transfusion practices in other developing countries (HPP 15 (2):217-22). PMID- 16689481 TI - Knowledge, attitudes and practices regarding sexually transmitted infections among general practitioners and medical specialists in Karachi, Pakistan. AB - OBJECTIVES: To determine the knowledge, attitudes and practices regarding diagnosis and treatment of sexually transmitted infections (STIs) among specialists that is, dermatologist, gynecologists and urologists and general practitioners (GPs) in Karachi, Pakistan. METHODS: Interviewers administered structured questionnaires to doctors conducting outpatient clinics at tertiary hospitals and/or private clinics in Karachi. All private clinics within a 10 km radius of the Aga Khan University and all tertiary hospitals having more than 100 inpatient beds were included in the study. RESULTS: One hundred doctors (54 specialists and 46 GPs) responded. Eighty doctors reported seeing at least one STI patient/month. The most commonly diagnosed STI the doctors reported was urethritis/cervicitis syndrome. 50% of the doctors knew the recommended antibiotics for gonorrhea though only 46% of these knew the correct dosage. Specialists were three times more likely to recognize the clinical presentation of herpes and twice as likely to treat chlamydia, syphilis and herpes with appropriate antimicrobials than GPs. 85% of the doctors advised their STI patients regarding condom usage; 36% thought that STI patients had loose sexual morals, 43% believed STI patients were drug addicts. Over 90% of the physicians were willing to attend educational sessions and follow a national STI treatment protocol. CONCLUSION: Doctors in Karachi, especially GPs, are deficient in appropriately managing and counseling STI patients. Among the specialists, urologists and dermatologists were more likely to manage STIs correctly than gynecologists. Karachi doctors should be educated in the correct management and counseling of STIs to prevent further spread of STIs including AIDS (Sex Transm Inf 2000;76:383-85). PMID- 16689482 TI - Multivariate analysis of risk factors associated with genital ulcer disease among incarcerated males in Sindh. AB - OBJECTIVE: To evaluate the potential risk behaviors associated with the lifetime risk of self reported genital ulcer disease (GUD) among prison inmates. SETTING: Prison inmates from 14 prisons of Sindh Province. METHODS: A cross-sectional study was conducted on 3395 prison inmates during July to December 1994. A questionnaire was used to assess the lifetime risk of self-reported GUD (whether or not the subject was ever affected with GUD up to present age) and to investigate demographic markers and risk behaviors for their possible association with lifetime risk of GUD using logistic regression analysis. RESULTS: The reported lifetime risk of GUD in the study sample was 11.4% (386/3395). In final multivariate logistic regression model the sexual behaviors which were independently associated with GUD were having sexual intercourse with female (adjusted OR = 1.7; 95% CI: 1.3-2.3, P=0.0002), sexual intercourse with a prostitute (adjusted OR = 1.5; 95% CI: 1.2-2.0, P=0.0008), sexual intercourse with man (adjusted OR = 2.2; 95% CI: 1.7-2.7, P=<0.001) and sexual intercourse with man during current incarceration (adjusted OR = 1.9; 95% CI: 1.2-2.9, P=0.0071). CONCLUSION: Health education needs to re-enforce monogamous relationship for high risk groups such as in our study. Although infrequent condom use was not a risk factor for GUD in this study, yet based on the results of previous studies, promotion of condom use should be the component of health education program (JPMA 50:115;2000). PMID- 16689483 TI - Survey and surveillance development in settings with low human immunodeficiency virus prevalence. AB - In most countries, during the early phases of a human immunodeficiency virus epidemic, independently initiated surveys of perceived high-risk groups tend to precede the development of formal surveillance systems. Unfortunately, in low prevalence settings, small sample sizes produce unreliable estimates of prevalence and trends, with an inevitable tendency towards positive results. In our study, we present sample size calculations and typical samples used in actual surveys, with Pakistan as our example. More useful data on risk behaviour and potential for spread can be derived from the study of commoner sexually transmitted diseases and associated risk behaviours, including assessments of knowledge, attitudes, beliefs and practices. PMID- 16689484 TI - Knowledge and misconceptions about sexually transmitted infections in married women--perspective from Islamabad. AB - OBJECTIVE: To study the knowledge of married women regarding existence of sexually transmitted infection (STI) their complications, treatment seeking, ways of preventing STI acquisition and opinion about sex education in schools/colleges and media. SETTING: The Mother and Child Health Center, a tertiary care hospital in Islamabad. METHODS: A cross-sectional survey based on sample of convenience was conducted, using a structured questionnaire with both close and open-ended questions. Trained women physician interviewers conducted the interviews after obtaining verbal consent. RESULTS: Out of 218 women approached for interviewing, only two refused to participate in the study. The mean age of the respondents was 28.5 years (range 18-53 years). One hundred and sixty-eight (77.8%) respondents had heard/knew about sexually transmitted infections (STIs). Two hundred and ten (97.2%) respondents had heard/ knew about AIDS and out of these 162 (77.1%) knew that it is also transmitted through sexual contact. One hundred and eighty two (84.2%) felt a need for sex education in the media, both print and electronic and 204 (94.4%) respondents said that they would like to learn more about sexually transmitted infections. One hundred ninety four (98.8%) respondents had heard the Latin term Leukorrhoea. Of these 158 (81.5%) thought it was a gynecological disease. Majority thought it caused weakness. CONCLUSION: Widespread misperceptions were found to exist, which does not augur well for the effective prevention of STIs in the country. Population based studies are required to study the knowledge and epidemiology of STIs, as well as a need for a health education campaign in the country (JPMA 51:389; 2001). PMID- 16689485 TI - Prevalence of HIV, hepatitis B and C amongst health workers of Civil Hospital Karachi. AB - OBJECTIVE: To determine the prevalence of HIV, Hepatitis B (HBV) and Hepatitis C (HCV) amongst the health workers of Civil Hospital Karachi (CHK). MATERIAL AND METHODS: Prospective study. A precoded Proforma was filled out which included questions regarding the knowledge, attitude and practices (KAP) of HIV, HBV and HCV. SETTING: Departments of a tertiary health care facility at CHK. Antibodies to HIV, HCV and Hepatitis B surface antigen (HBsAg) were done using enzymes linked immunosorbent assay [ELISA]. RESULTS: Uptake of screening was 98% to those offered. The prevalence was 5.6% for antibodies to HCV, 2.4% for HBsAg, while none of those studied had antibodies to HIV. CONCLUSION: Our results show the prevalence of antibodies to HCV in health workers are 20 folds higher than health workers in the developed countries. Similarly, the prevalence of HBV although not as high as HCV is significant. Seroprevalence of HIV does not exist in this group. We need to ensure better training; regulations regarding preventive and safety measures also need to be enforced (JPMA 52:92; 2002). PMID- 16689486 TI - Community knowledge, attitude and practices regarding sexually transmitted infections in a rural district of Pakistan. AB - OBJECTIVE: To determine knowledge, attitude and practices regarding sexually transmitted infections in district Khairpur, Sindh, Pakistan. DESIGN AND METHODOLOGY: A needs assessment study employing a community based survey, key informant interviews and focus group discussions was conducted in all seven sub districts of the district. This paper is based on the results of the key informant interviews and the focus group discussions. Thirty interviews and six group discussions were held with members of the community and with the health care providers. RESULTS: There was little awareness regarding causes and prevention of sexually transmitted infections in the community. The situation was slightly better among health care providers. While health care providers believed that the prevalence of sexually transmitted infections is high, the community did not consider themselves at risk. The community believed that these diseases are a problem among a sub-population of male adolescents, especially those who have homosexual relations. However, due to social norms, they rarely discussed such health problems with other family members or elders. Adolescents with any sexual health problems visit hakims and quacks. The community was aware about aetiology and some of the risk factors associated with AIDS and hepatitis, most probably due to the recent public health campaigns against both diseases. CONCLUSION: Considering the suspected high prevalence of sexually transmitted infections and the relative lack of knowledge, it is imperative that a public health intervention be initiated. This must include educating not only the community but also the health workers. The governments' initiative to train community workers in reproductive health is a step in the right direction (JPMA 52:21, 2002). PMID- 16689487 TI - Socio-economic and demographic factors associated with injecting drug use among drug users in Karachi, Pakistan. AB - OBJECTIVE: To identify the socio-economic and demographic factors associated with injecting drug users (IDUs) in Karachi. METHODS: We recruited 242 IDUs (taking drugs through sub-dermal routes) and 231 non-IDUs (taking drugs other than sub dermal routes) from February through June 1996. IDUs were interviewed regarding sociodemographic factors, economic condition, and social network (marital status, living with spouse). In addition, information regarding location of drug users within the city (districts of Karachi) and current history of sexually transmitted diseases (STDs) were gathered. Moreover, blood samples were also obtained for HIV testing. RESULTS: Multivariate analysis showed that the income generation via illegal modes [AOR 1.6, 95% CI 1.0-2.6], non-sharing of income with family (AOR 1.7, 95% CI 1.1-2.7) and presence of suicidal thoughts (AOR 2.0, 95% CI 1.1-3.8) were associated with the use of drugs through injections. Further, drug users from districts West, East and Central were more likely to use drug through injection as compared to drug users from district South. The history of genital herpes was also found to be associated with injecting drug use. One IDU was found seropositive for HIV. CONCLUSION: The high-risk behaviors, such as illegal modes of earning and presence of suicidal thoughts, among IDUs suggest that the group needs rehabilitation programme. Moreover, non-sharing of income suggest that IDUs are isolated from social network, therefore primary prevention activities with focus on improving socio-economic conditions and social networking can reduce drug use through injections. Focused interventions on target districts would be helpful in reducing IDU (JPMA 53:511; 2003). PMID- 16689488 TI - Profile of men suffering from sexually transmitted infections in Pakistan. AB - BACKGROUND: To evolve effective prevention and efficient treatment strategies for sexually transmitted infections (STIs) in a country, comprehensive understanding of the prevalent STI and their modes of transmission is needed. The aim of this present study was to generate such data for Pakistan. METHODS: The study was conducted between June 1999 and September 1999 in four provincial capitals. In each city, data was collected from one or more teaching hospitals and a number of general practitioners. During this period, 465 men suffering from STIs were interviewed. RESULTS: The mean age of study population was 31.9 +/- 8.6 years, 60% were married, 58.3% urbanites, 81.7% living with their families and 83.3% smokers. Only 10.5% were drug addicts. Out of 465 cases, 27.5% cases had gonorrhea, 31.6% syphilis, 17.2% chancroid, 18.3% herpes, 5.2% chlamydial infections while only one case (0.2%) was HIV positive. Most men (55%) acquired the infection heterosexually, 11.6% through homosexuality, 18.4% through bisexual relations and 2 men (0.5%) reported bestiality. Among 78.1% of those contracting the infection heterosexually, the source of infection was a sex worker. None acquired infection through his wife. The knowledge about STIs was very poor. Wrong notions were prevalent. CONCLUSIONS: The pattern of STIs is different from developed countries. Those acquiring STIs in Pakistan were older, married and living with their families whereas in developed countries most men acquiring STIs are younger, unmarried and living alone. PMID- 16689489 TI - High-risk sexual behaviours among drug users in Pakistan: implications for prevention of STDs and HIV/AIDS. AB - Our objective was to describe HIV/STD risk behaviours and awareness among a community-based sample of drug users in Pakistan. Drug users contacted through street outreach by a non-governmental organization in Quetta, Peshawar and Rawalpindi underwent interviewer-administered questionnaires. Descriptive statistics were used to characterize sexual behaviours by city, marital status and the use of injection drugs. Logistic regression was used to identify correlates of ever having an STD. Of 608 drug users studied, all but one was male; median age was 32 years and 45% had no formal education. Half were married, of whom 25% were living with their wives. Sexual behaviours were reported as follows: 14% had sex with other males, 28% reported sex with males and females, 49% had paid money to have sex and only 10% had ever used condoms. One-fifth reported having had an STD and about 40% reported having suffered from either one or more STD-related symptoms. Only 41% had heard about HIV/AIDS, of whom 17% knew that HIV/AIDS could be transmitted through sexual contact. In conclusion, high risk sexual behaviours are prevalent among male drug users in Pakistan, and awareness of transmission risks is low. These data attest to the urgent need for effective and specific interventions in Pakistan to prevent transmission of HIV and STDs among drug users and their sex partners (I J STD & AIDS 2004;15:601-7). PMID- 16689490 TI - High frequency of false positive results in HIV screening in blood banks. AB - BACKGROUND: This study was carried out to determine the frequency of false positive results during serological screening for the presence of antibodies against HIV-I/2 in blood banks. METHODS: A cross-sectional study was conducted from January-December 1999 as screening of voluntary non-renumerated blood donor pool for HIV in the public sector blood banks, in all the six divisions of Balochistan. 5000 subjects were screened for the presence of antibodies against HIV-I/ 2. The subjects were all males between the age group 18-50 years, attending the public sector blood banks as non-renumerated blood donors. Strategy 1 was adopted for initial screening. Strategy II and III were observed in retesting on ELISA, as recommended by UNAIDS/WHO for blood banks. RESULTS: Out of 5000 subjects, 48 (0.96%) were positive for HIV-I/2 on Strategy I, 37 (77% of 48) met the criteria of false positive, while only 11 (0.22% of 5000) were found to be true positive. CONCLUSION: In blood banks, screening for HIV antibodies is performed for intervention of the positive donations. UNAIDS / WHO Strategy-I is observed on a smaller workload blood banks where donations are less than 20 per day. A high rate of false positive results in serological HIV screening on Strategy-I depicts that the test is highly sensitive but not highly specific. Labeling someone with HIV positive, when actually he is not, forces the health authorities to find other ways of HIV screening in blood banks, which should be much more specific and therefore reliable. PMID- 16689491 TI - Prevention and control of HIV/AIDS among injection drug users in Pakistan: a great challenge. PMID- 16689492 TI - An outbreak of HIV infection among injection drug users in a small town of Pakistan: potential for national implications. PMID- 16689493 TI - [Managerial knowledge required of the nurse in the family health program]. AB - This article is a part of a study entitled "Managerial skills required of the nurse in the Family Health Program" which sought to identify the managerial knowledge required of nurses, to run the nursing services in the Family Health Units in a city on the coast of Santa Catarina. Taking the perspective of professionals working in these units, the study used the descriptive-exploratory method, with a quantitative approach and a semi-structured questionnaire. The study sample consisted of 72 participants, who attributed a score of between 1 and 5 for each item on the questionnaire. The results indicated an emphasis on the mastery of a good knowledge of SUS (the national Brazilian healthcare system) policies, which would lead to the development of skills by the nurses, enabling them to achieve a good performance and consequently, ensuring quality care and the concrete resolution of multiple problems, guaranteeing satisfaction for the community, a decrease in levels of epidemiology and a better quality of life for the population. PMID- 16689494 TI - [Breast-feeding experience for women workers and students from a public university]. AB - This qualitative research aimed to know the main interfering elements in the breast-feeding process as experienced by professional women and by students, that was carried out with 65 professional women and students from a public university in Sao Paulo state. The data collection was proceeded by interviews which contents were analyzed according to Taylor and Bogdan and Symbolic Interactionism approaches. Results indicated that the breast-feeding process for these women demonstrated to be conditioned and highlighted by the conditions the women encounter in their domestic, professional, and study settings. The physical setting and the relations among their relatives, superiors and peers exerts a strong influence on their determination to keep on breastfeeding. PMID- 16689495 TI - [Family caregivers' perception about a program of hospital discharge]. AB - The preparation of hospital discharge, seen as a strategy in Health Education, may help the family to continue household care after hospital discharge. This descriptive study with qualitative approach aims at analyzing how the family caregiver who assists elderly patients with CVA (cerebrovascular accident) sees a discharge planning program in a university hospital. Twelve family caregivers were interviewed. Data were analyzed according to the Content Analysis Technique, and four categories emerged: opinion about the program; adequacy of guidance according to the demands for care; improvement points; and repercussions of the caregiver. It is suggested the implementation of similar programs in health care services regarding the elderly patient with CVA and his/her family. PMID- 16689496 TI - [The senior students perception about basic nursing tools and their applicability]. AB - This paper presents a descriptive exploratory study of student's perception about the applicability of basic nursing tools on the daily professional practice. Results highlights the factors that contribute to the continuity of dichotomy between theory and practice and the possibilities of overcoming/transformation of this reality. In addition, the study shows that the concretization of the know how in nursing requires from each nurse to be aware of his/her personal, professional and social responsibility to offer a high quality health care, through the full professional know-how and knowledge. PMID- 16689497 TI - [Nursing diagnoses identified in pregnant patients under prenatal care]. AB - This descriptive research aimed to analyze the Nursing Diagnoses identified in low-risk pregnant patients. Eleven pregnant women received prenatal care in a safe maternity in the state of Goias. The data were classified under the NANDA (North American Nursing Diagnosis Association) taxonomy II and analyzed according to the referrals in obstetrics. Twenty-five different Nursing Diagnoses were identified. All pregnant women presented: insufficient knowledge (different levels and subjects), disturbed sleep pattern, activity intolerance, self-care deficit (bathing and hygiene), stress urinary incontinence, and some risk diagnoses. Identifying NANDA's Nursing Diagnoses allows the identification of the risk level, early detection of complications, and individualization in nursing assistance. PMID- 16689498 TI - [Nursing and family planning: the interfaces of contraception]. AB - This work analyses some aspects of family planning policy in Brazil related to contraceptive methods as published in Revista Brasileira de Enfermagem between 1960 and 1997. The subject analysis considers three categories: Catholic church, feminist and official state discourse. According to the records, in the decade of 1960 the Catholic church discourse was predominant. This situation continued in the decade of 1970, accompanied with emerging voice of the official state discourse. In the decade of 1980, the feminist discourse joined. Finally, in the decade of 1990 the feminist discourse continued, the official state discourse was consolidated and that of the Catholic church was not found any more. In conclusion, the journal established family planning on the agenda of Brazilian nursing by the means of scientific production. The methodology used enabled us to identify what are the obligations of nursing with respect to family planning in the context of its relationship with state policies and with the society. PMID- 16689499 TI - [Communication: a percieved need in the pre-operative period for surgical patients]. AB - The study had as objective to evaluate the patients' feelings in the pre operative period. The study is descriptive with qualitative approach. It was accomplished in a surgical unit of a public hospital of Sao Paulo. The population was constituted of patients in the pre-operative period, in the age group from 10 to 80 years. Non-systematized observation and semi-structured glimpses were used for data gathering. The results showed the lack of effective communication caused by the obstacle disturbs the therapeutic relationship, leading the patients to anxiety and depression during the hospitalization, because the lack of surgery orientation and support by the health team. PMID- 16689500 TI - [Teaching of obstetrical nursing in Brazil: successes and failures from 1972 to 1996]. AB - Investigation with descriptive and qualitative approach. Political-social context of Nurse teaching are analyzed, with emphasis in policies formation. Teaching programs of Obstetrical Nursing from 1972 to 1996 utilized by five graduate teaching schools were described and analyzed according to geographical region, for dialetic analysis of thematic and contents. It was evidenced that disciplines has given priority for governmental purposes centered on pregnant-puerperal cycle, although it should had some political-social movements influence, mainly feminist and progressive sanitation, on which the PAISM is based. The emerging content of these movements were not exposed (family health, subjectivity, violence, gender, among others). Most of institutions were on curricular reformulation to attend the 314/94 statement which determines generalist nurse curriculum in Brazil. PMID- 16689501 TI - [Family history of children with chronic kidney failure: data collection]. AB - The systematization process of care has as its first step the nursing history. This study aims to present the experience of data collection for care planning involving families of children with Chronic Kidney Failure on peritoneal dialysis. Family Nursing was used as a theoretical reference framework. Data were collected by means of in-depth interviews, documentary analysis, field diary, genogram and ecomap. Study participants were four families. The authors discuss in-depth interviews, its limitations and advantages, the construction of the genogram and ecomap and ethical, legal and social implications. Finally, it is presented general considerations on how to apply these instruments in care delivery involving families of children with Chronic Kidney Failure on peritoneal dialysis. PMID- 16689502 TI - [Incidence and features of cesarean section and natural childbirth: study in a city from Ceara's countryside]. AB - The research aimed to describe the features of a given population studied, as civil state, occupation and number of pre-natal appointments; to verify the incidence of cesarean section and normal labor in clients attended in an institution; to compare these occurrence in the period of research, discussing the indications of cesarean sections more frequently referred. It is a descriptive-retrospective study based upon patient records from a philanthropic institution of the National Health System. It highlighted a rising percentage of cesarean sections with high incidence in teenagers. It evidenced that the main indications of cesarean section were due to former cesarean section and pregnancy specific hypertensive disease. It was recorded predominance of pregnants with 4 to 7 pre-natal appointments. Changes in pre-natal care can impact in cesarean section rate, preparing the mother to natural labor, something that can be closely related to midwives obstetrical nurses and physicians capacitation, as well as the placement of group care instead individual care. PMID- 16689503 TI - [Climacteric: the corporeity as cradle of life experience]. AB - Qualitative study of phenomenological approach, accomplished from women's testimony aged between 58 and 55, who had presented spontaneous menopause over the last 12 months or more. The results evidenced five thematic categories that were analyzed and interpreted under the light of Maurice Merleau-Ponty's philosophical theoretical referral. Among the categories that have emerged from the testimonies, the unveiled theme "Make yourself aware of the world through your body in space and time" will be presented with the objective of stimulating the reflection on existential aspects of experiencing this reproductive cycle phase and make considerations on the climacteric woman health care. PMID- 16689504 TI - [Factors that interfere in the parturient women's behavior: focus on ethnonursing]. AB - This study aimed to understand the behavior of pregnant women in labor and delivery. Seven parturient women who had their children through normal delivery in a public maternity in Fortaleza, Ceara. Data collection was carried out in June to August, 2003. The research method used for the collection and analysis of data was the etnonursing. The data studied showed us through the categories: experiencing pregnancy and assistance in the maternity. We concluded that there are several factors that justify the woman's behavior during the experience of labor and delivery and that the health professionals' assistance should be always aimed at the humanistic care linking cultural and scientific knowledge. PMID- 16689505 TI - [Expression of needs in the field of the basic care in sexual health]. AB - This article approaches the ethical-political meanings present in basic attention to sexual and reproductive health in Cuiaba-MT. Aspects related to the contradictory expression of needs and demands in the field of the sexuality are argued. Its correlation with the historical forms of attention to the health is emphasized, also considering the social and cultural character of these interpretations and practices in health and the sexuality. The relevance of a wide understanding of needs in the field of sexuality to the construction of projects of integral attention is approached, considering the way these needs are presented in a given health service. PMID- 16689506 TI - [Construction of optometric scales in the light of philosophy]. AB - It is by walking that the new arises. This study aims at establishing new issues in the construciton of optometric scales of pictures accordingly to their shape and content. It is a reflection (bibliographic-analytical) based on Danta's dissertation. It is observed that the matter is the optonometric scale ready for use. The shape, in turn, is what was developed until the generation of the matter, the optical calculations and the law of refraction, apart from the ocular anatomy and physiology. The new aspect developed is culture. The philosophical question shows the infinity of knowledge and reveals that the view of certain research at a certain time can be rebuilt in another. PMID- 16689507 TI - [Understanding King's model on the paradigm of symbolic interactionism]. AB - It was aimed to reflect on King's Theory, according to the approach of Symbolic Interaccionism, and theory analyses model of Meleis. To reach the objective, we proceeded the reading of the three models as mentioned before, looking for the consistencies and discrepancies among the concepts and correlating them. The study allowed to conclude that the theories agree when elucidating the man as a being that reacts and search to understand the meaning of things to his/her circuit, drifting and judging their actions and the one of the other ones. It is important the subject of the meanings could be modified and they generate the elaboration of goals in common. PMID- 16689508 TI - [Nursing interventions for the most frequent complications during hemodialysis procedure: literature review]. AB - Hemodialysis procedure has potential complications and nurses must be able to intervent on them. Considering this context, this article aims to describe the most frequent hemodialysis-related complications as well as nursing-related interventions. This bibliographic review consists in LILACS, BDENF and MEDLINE search using pertinent descriptors. Results demonstrated that major complications are: hypotension, hypertension, cramps, nausea and vomiting, headache, chest and low back pain, scratching, fever and shriving. For these complications there are specific nursing interventions that can be delivered. It was concluded that nurse's role is essential for monitoring, identification, and intervention in such complications and that this role also is a differential to obtain quality in hemodialysis procedure. However, there is a need to perform more research in nursing area to better define the nurse's role. PMID- 16689509 TI - [Florence Nightingale and charity sisters: revisiting the history]. AB - This study presents an historical analysis on the links between the nursing practice and the influence received from various religious orders/associations along the times, especially from Saint Vincent Paul's charity sisters. The professional nursing which was pioneered by Florence Nightingale in the XlXth century, was directly influenced by the teachings of love and fraternity. In addition, other contributions from the religious orders/associations were the concepts of altruism, valorization of an adequate environment for the care of patients, and the division of work in nursing. The study shows the influence of Charity Sisters on Florence Nightingale. PMID- 16689510 TI - [Outsourced services and their consequences for nursing care]. AB - This experience report intends to turn visible and to share with other professionals a practical experiment. The influence of outsourced services is emphasized in the process of nursing care, focusing the difficulties found, in the same institution, in the units not outsourced related to nursing care and problem resolution of clients submitted to hemodynamics procedures that are hospitalized after the intervention and/or return to common units of the Institution which services are outsourced. The experience evidences the need of a integrated team between the two services, using the continuing education as a way to rescue fundamental knowledge for clients' integral care, as well as a more committed profession with the being and making nursing. PMID- 16689511 TI - [Ethics in care during the dying process: case report]. AB - This study considers ethical issues related to the terminally ill. Its aims are to report the experience of home care and to promote a reflection about the ethical dilemma of caring the terminally ill. We concluded that the bioethical acting in this kind of situation involves a kind of conscience that can only be developed when the patient's reality is understood. We also realized the relevance of an interdisciplinary approach as a strategy for the holistic care to the terminally ill and the need to give attention to care when the cure is not possible, but the quality of life in the last moments of life. PMID- 16689512 TI - Satellite clinics--"cherry picking" in a community orthopedic practice. PMID- 16689513 TI - Are you being bribed? Health care ethics and compliance in the AdvaMed Code era. Part II. PMID- 16689514 TI - Femoral shaft fractures: is rigid intramedullary nailing safe for adolescents? AB - Despite recent reports of avascular necrosis of the femoral head after intramedullary nailing in children, this fixation method is still being advocated for the treatment of femoral shaft fractures. In this article, we report a case of early-onset, severe, symptomatic avascular necrosis of the femoral head after intramedullary nailing in a 13-year-old boy. Given the possibility of this devastating complication, surgeons should consider the risk-benefit ratios of various techniques for managing femoral shaft fractures in adolescents. PMID- 16689515 TI - Lumbar degenerative classification study. AB - The purpose of this study was to develop and validate a classification of indications for fusion in lumbar degenerative disease. Nineteen spine surgeons reviewed a series of 32 case histories and selected the indication for fusion based on an outlined classification system. To determine the degree of interrater variability, K coefficients were calculated (K for all 32 cases, 0.63). Results from this study show the significant difficulty in classifying the indication for fusion in lumbar degenerative disease. The level of the 19 surgeons' agreement regarding surgical indication was only moderate, despite a study design that eliminated controversial issues of patient and procedure selection. To a significant extent, the difficulty in classifying indication for fusion underlines the importance of the process. If we cannot agree on why a specific patient is selected for fusion, it is then impossible to accurately compare outcomes for a given disease process or surgical technique. For this reason, an ongoing effort to refine nomenclature and classification is necessary. PMID- 16689516 TI - Functional capacity before and after simulated thumb carpometacarpal joint fusion. AB - Lateral pinch strength, grooved pegboard test time, and 22 activities of daily living (ADLs) were assessed in 10 asymptomatic patients initially without and then with splinting of the thumb carpometacarpal (CMC) joint (simulating thumb CMC fusion). Before immobilization, all patients rated a perfect total score of 88 on the 22 ADLs. After immobilization, mean total score dropped to 78.3, with particular difficulty noted in manipulating money, cutting meat, turning a doorknob, using a can opener, using scissors, and writing. Lateral pinch strength decreased from 22.4 lb without immobilization to 20.8 lb after splinting. Mean grooved pegboard test time increased from 53.1 to 58.3 seconds with immobilization. Thumb CMC fusion is not without particular functional defects. Patients' understanding of these difficulties allows for informed decisions regarding whether surgical treatment for thumb CMC arthritis is desired. PMID- 16689517 TI - Intrauterine vascular deficiency secondary to methylenetetrahydrofolate reductase deficiency: 2 case reports. PMID- 16689518 TI - A carpometacarpal joint ganglion cyst causing median neuropathy. PMID- 16689519 TI - Pectoralis major ruptures. AB - Although previously considered rare, ruptures of the pectoralis major muscle and tendon have become more common over the past 20 years with increasing participation in competitive sports and weight lifting. These injuries result from maximal eccentric contraction of the muscle with the arm held in an abducted, extended position. Most often, they occur near the tendon insertion. Complete ruptures are more common than partial tears, though many mild strains and partial tears likely go unreported or undiagnosed. Magnetic resonance imaging is the diagnostic study of choice when the diagnosis or extent of the injury is unclear. Partial tears and neglected complete tears with near complete restoration muscle strength after rehabilitation may be treated nonoperatively, with good results. Excellent results can be obtained with early anatomic repair for complete ruptures, though good results have also been reported with delayed repair of complete injuries. Early diagnosis and, when appropriate, anatomic surgical repair are the factors critical to optimizing patient outcomes after pectoralis major rupture. PMID- 16689520 TI - The detection of nitro pesticides in mainstream and sidestream cigarette smoke using electron monochromator-mass spectrometry. AB - Over the past decade, electron monochromator-mass spectrometry (EM-MS) has been shown to be a selective and sensitive technique for the analysis of a wide variety of electrophilic compounds in complex matrixes. Here, for the first time, three different dinitroaniline pesticides, flumetralin, pendimethalin, and trifluralin, have been shown to be present in both mainstream and sidestream tobacco smoke using an EM-MS system. A number of cigarettes were tested that included three pure-tobacco-type cigarettes, an experimental reference cigarette, and 11 commercial cigarettes. Due to the complexity of the smoke particulate matter, the pesticides were identified only after each sample was subjected to a multistep cleanup process that included phenyl solid-phase extraction; an acid wash; aminopropyl solid-phase extraction; and finally, normal phase LC fractionation. All cigarette types tested showed the presence of the three pesticides in the tobacco smoke, with flumetralin ranging from trace levels up to 37 (+/- 9) ng/cig, pendimethalin ranging from trace levels up to 10.4 (+/- 0.6) ng/cig, and trifluralin ranging from trace levels up to 47 (+/- 17) ng/cig. Acute toxicity information is presented for the three pesticides. PMID- 16689521 TI - Magnetic and gold-coated magnetic nanoparticles as a DNA sensor. AB - In this study, we report the chemical synthesis and functionalization of magnetic and gold-coated magnetic nanoparticles and the immobilization of single-stranded biotinylated oligonucleotides onto these particles. Selected sequences specific to the BRCA1 gene were used as a test platform. The binding of oligonucleotides to these particles was achieved through a streptavidin-biotin bridge via a carbodiimide activation protocol. Particle size and oligonucleotide attachment were confirmed by transmission electron microscopy; oligonucleotide binding was characterized by Fourier transform infrared spectroscopy and hybridization confirmed by fluorescence emission from the fluorophore attached to the target oligonucleotide strand. The rate of hybridization was measured using a spectrofluorometer and a microarray scanner. The rate of hybridization of oligonucleotides bound to the synthesized particles depends on the inorganic support material and its surface chemistry. The rate of hybridization increased concomitantly with the concentration of the probe and the target in the reaction medium. Furthermore, exposure of probe and target oligonucleotide to a combination of target and noncomplementary DNA strand reduced the rate of hybridization, possibly because of steric crowding in the reaction medium and cross-linking between reacting oligonucleotides and the noncomplementary strands. The study undertaken opens several possibilities in bioconjugate attachment to functionalized iron and iron nanocomposite structures for controlled manipulation and handling using magnetic fields. PMID- 16689522 TI - Method for measuring thickness of dielectric films using microdielectric fringe effect sensors. AB - A method for noninvasive thickness measurements of dielectric films using fringe effect (FE) sensors is developed and experimentally validated. The fringing electrical field, created by electrodes microfabricated at the film substrate, depends on the film thickness and dielectric permittivity of the film under test (FUT). The unknown film thickness is estimated by matching the theoretical prediction of thickness-dependent sensor admittance with the measured value. In the case of FE sensors with spatially periodic, interdigitated electrode (IDE) configuration, the admittance prediction is simplified, which allows for the real time measurements of changing thickness. The developed method can be used to continuously measure the changing dielectric permittivity of the FUT material, which makes it possible to determine the thickness of films of changing dielectric properties, caused by chemical or other transformations. The application of the developed method is demonstrated experimentally by measuring the thickness of silicon nitride film deposited in several increments on the quartz substrate of the IDE sensor. In the expected range of sensor sensitivity, the results show an excellent agreement with the independent thickness measurements. PMID- 16689523 TI - UV embossed polymeric chip for protein separation and identification based on capillary isoelectric focusing and MALDI-TOF-MS. AB - This paper demonstrates a ultraviolet (UV)-embossed polymeric chip for protein separation by capillary isoelectric focusing (CIEF) and identification by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The polymeric chip was replicated by a UV-embossing technique using a soft rubber mold. Five diverse widely investigated families of UV-cured formulations were examined for MALDI ionization efficiency of bovine serum albumin (BSA) samples spotted on the polymer surfaces. The signal and signal-to-noise ratio from the polyester formulation were each 12 times those obtained with PMMA (a commonly used polymer material for MALDI) at picomole sample concentration. A polyester chip was successfully used to carry out CIEF to separate proteins, followed by MALDI-TOF-MS identification. Issues related to the successful chip fabrication and protein separation and identification are discussed. PMID- 16689524 TI - Detection of albumin unfolding preceding proteolysis using Fourier transform infrared spectroscopy and chemometric data analysis. AB - The hydrolysis of bovine serum albumin with protease K at 60 degrees C has been studied by means of infrared spectroscopy. Two-dimensional correlation spectroscopy (2DCoS) has been used to study spectral changes in the reaction. The use of the multivariate curve resolution-alternating least-squares method applied to infrared measurements allowed the recovery of pure infrared spectra and concentration profiles of the different species involved in the reaction. Special attention was paid to the careful inspection of residuals again using 2DCoS. In this way, a heat-induced unfolding step previous to protein hydrolysis was identified. The infrared spectra of the intermediate species showed a more disordered structure than native albumin, the decrease in alpha-helix conformation being especially noticeable. The formation of beta-sheet aggregates due to heating was detected too. PMID- 16689525 TI - Virus electrodes for universal biodetection. AB - A dense virus layer, readily tailored for recognition of essentially any biomarker, was covalently attached to a gold electrode surface through a self assembled monolayer. The resistance of this "virus electrode", Z(Re), measured in the frequency range from 2 to 500 kHz in a salt-based pH 7.2 buffer, increased when the phage particles selectively bound either an antibody or prostate specific membrane antigen (PSMA), a biomarker for prostate cancer. In contrast to prior results, we show the capacitive impedence of the virus electrode, Z(Im), is both a noisier and a less sensitive indicator of this binding compared to Z(Re). The specificity of antibody and PSMA binding, and the absence of nonspecific binding to the virus electrode, was confirmed using quartz crystal microbalance gravimetry. PMID- 16689526 TI - Development of recombinant-based mass spectrometric immunoassay with application to resistin expression profiling. AB - This report addresses the need for additional assays for human resistin (hRES) by developing a rational progression of the mass spectrometric immunoassay to incorporate recombinant proteins. The recombinant-based hRES mass spectrometric immunoassay (RES-MSIA) was initially developed for the qualitative analysis of the human resistin homodimer from normal (healthy) plasma samples. The method involved selective extraction and detection of both endogenous and recombinant resistant proteins. RES-MSIA was then applied to the rigorous quantification of resistin. The resistin standard addition curve was constructed from serially diluted concentrations of rhRES using endogenous hRES, inherent in the human plasma, as the internal reference standard (IRS). The roles of endogenous and recombinant resistin were subsequently reversed, using rhRES as the IRS during RES-MSIA quantification. Concurrently, the relative ratio of hRES to rhRES was used as an ancillary technique to rapidly determine the relative concentration of hRES in each of plasma samples. Overall, normal hRES levels determined by RES MSIA were found to be comparable to those selected and determined by ELISA. With regard to gender, female donor samples were slightly elevated over males. Four single cardiac samples were analyzed and found to have hRES concentrations approximately three times that of the normal. The recombinant-based RES-MSIA is rapid and is amendable to parallel high-throughput robotic processing of resistin related disease cohorts. PMID- 16689527 TI - Determining the conformation of thiolated poly(ethylene glycol) on Au nanoshells by surface-enhanced Raman scattering spectroscopic assay. AB - The packing density of thiolated poly(ethylene glycol) (PEG) adsorbates on Au nanoshells is determined by exploiting the surface-enhanced Raman scattering response of individual nanoshell substrates. By incorporating the linker molecule p-mercaptoaniline (pMA), the number of 2000 MW and 5000 MW PEG molecules on each nanoparticle is determined by interpolation of the Langmuir isotherm for pMA. We conclude that both PEG adsorbates maintain a compact "brush" rather than an extended "mushroom" configuration on nanoshell surfaces. PMID- 16689528 TI - Gold and silica-coated gold nanoparticles as thermographic labels for DNA detection. AB - The infrared emissivity of Au and silica-coated Au nanoparticles (Au NPs) deposited on indium tin oxide substrates was investigated. NPs were irradiated with laser light at a frequency close to the Au plasmon resonance band, and the blackbody radiation emitted as a result was monitored with an IR camera equipped with an InAs array detector. The differences in temperature before and after laser irradiation were recorded (T-jumps) and were found to be directly proportional to the number of particles present on the slide and to the laser power used in the experiment. Coating Au NPs with silica increased the measured T jumps 2-5 times, depending on the thickness of the silica shell. This was in agreement with the observation that silica has a much higher IR emissivity than Au. Both Au and silica-coated Au NPs were then tested as labels for thermographic DNA detection. Target DNA concentrations as low as 100 pM were recorded when Au NPs were used as labels and as low as 10 pM when silica-coated Au NPs were used. PMID- 16689529 TI - Nonlinear data alignment for UPLC-MS and HPLC-MS based metabolomics: quantitative analysis of endogenous and exogenous metabolites in human serum. AB - A nonlinear alignment strategy was examined for the quantitative analysis of serum metabolites. Two small-molecule mixtures with a difference in relative concentration of 20-100% for 10 of the compounds were added to human serum. The metabolomics protocol using UPLC and XCMS for LC-MS data alignment could readily identify 8 of 10 spiked differences among more than 2700 features detected. Normalization of data against a single factor obtained through averaging the XCMS integrated response areas of spiked standards increased the number of identified differences. The original data structure was well preserved using XCMS, but reintegration of identified differences in the original data reduced the number of false positives. Using UPLC for separation resulted in 20% more detected components compared to HPLC. The length of the chromatographic separation also proved to be a crucial parameter for a number of detected features. Moreover, UPLC displayed better retention time reproducibility and signal-to-noise ratios for spiked compounds over HPLC, making this technology more suitable for nontargeted metabolomics applications. PMID- 16689530 TI - Aptameric enzyme subunit for biosensing based on enzymatic activity measurement. AB - The aptameric enzyme subunit (AES), which is a DNA aptamer composed of an enzyme inhibiting aptamer and a target molecule-binding aptamer, has been developed for the biosensing of target molecules. We used a thrombin-inhibiting aptamer as the aptamer that inhibits enzymatic activity. The thrombin-inhibiting aptamer folds into the G-quartet structure, which plays an important role in its inhibitory activity. As a target molecule-binding aptamer, an adenosine-binding aptamer was inserted into the G-quartet structure of the thrombin-inhibiting aptamer to enable the change of the G-quartet structure upon the recognition of adenosine. In the present study, the change in the G-quartet structure led to a change in the thrombin inhibition activity, and adenosine was successfully detected by measuring the thrombin activity in a homogeneous solution without bound/free separation. We constructed two kinds of AESs; one of the structures is universal and can be used for designing any target molecule-binding aptamer. Since the enzyme activity is measured, AESs enable the simple and high-sensitivity detection of target molecules in a homogeneous assay. PMID- 16689531 TI - Characterizing the structures and folding of free proteins using 2-D gas-phase separations: observation of multiple unfolded conformers. AB - Understanding the 3-D structure and dynamics of proteins and other biological macromolecules in various environments is among the central challenges of chemistry. Electrospray ionization can often transfer ions from solution to gas phase with only limited structural distortion, allowing their profiling using mass spectrometry and other gas-phase approaches. Ion mobility spectrometry (IMS) can separate and characterize macroion conformations with high sensitivity and speed. However, IMS separation power is generally insufficient for full resolution of major structural variants of protein ions and elucidation of their interconversion dynamics. Here we report characterization of macromolecular conformations using field asymmetric waveform IMS (FAIMS) coupled to conventional IMS in conjunction with mass spectrometry. The collisional heating of ions in the electrodynamic funnel trap between FAIMS and IMS stages enables investigating the structural evolution of particular isomeric precursors as a function of the intensity and duration of activation that can be varied over large ranges. These new capabilities are demonstrated for ubiquitin and cytochrome c, two common model proteins for structure and folding studies. For nearly all charge states, two-dimensional FAIMS/IMS separations distinguish many more conformations than either FAIMS or IMS alone, including some with very low abundance. For cytochrome c in high charge states, we find several abundant "unfolded" isomer series not distinguishable by IMS, possibly corresponding to different "string of beads" geometries. The unfolding of specific ubiquitin conformers selected by FAIMS has been studied by employing their heating in the FAIMS/IMS interface. PMID- 16689532 TI - Surface coverage and structure of mixed DNA/alkylthiol monolayers on gold: characterization by XPS, NEXAFS, and fluorescence intensity measurements. AB - Self-assembly of thiol-terminated single-stranded DNA (HS-ssDNA) on gold has served as an important model system for DNA immobilization at surfaces. Here, we report a detailed study of the surface composition and structure of mixed self assembled DNA monolayers containing a short alkylthiol surface diluent [11 mercapto-1-undecanol (MCU)] on gold supports. These mixed DNA monolayers were studied with X-ray photoelectron spectroscopy (XPS), near-edge X-ray absorption fine structure spectroscopy (NEXAFS), and fluorescence intensity measurements. XPS results on sequentially adsorbed DNA/MCU monolayers on gold indicated that adsorbed MCU molecules first incorporate into the HS-ssDNA monolayer and, upon longer MCU exposures, displace adsorbed HS-ssDNA molecules from the surface. Thus, HS-ssDNA surface coverage steadily decreased with MCU exposure time. Polarization-dependent NEXAFS and fluorescence results both show changes in signals consistent with changes in DNA orientation after only 30 min of MCU exposure. NEXAFS polarization dependence (followed by monitoring the N 1s --> pi* transition) of the mixed DNA monolayers indicated that the DNA nucleotide base ring structures are oriented more parallel to the gold surface compared to DNA bases in pure HS-ssDNA monolayers. This indicates that HS-ssDNA oligomers reorient toward a more-upright position upon MCU incorporation. Fluorescence intensity results using end-labeled DNA probes on gold show little observable fluorescence on pure HS-ssDNA monolayers, likely due to substrate quenching effects between the fluorophore and the gold. MCU diluent incorporation into HS ssDNA monolayers initially increases DNA fluorescence signal by densifying the chemisorbed monolayer, prompting an upright orientation of the DNA, and moving the terminal fluorophore away from the substrate. Immobilized DNA probe density and DNA target hybridization in these mixed DNA monolayers, as well as effects of MCU diluent on DNA hybridization in complex milieu (i.e., serum) were characterized by surface plasmon resonance (SPR) and (32)P-radiometric assays and reported in a related study (Gong, P.; Lee, C.-Y.; Gamble, L. J.; Castner, D. G.; Grainger, D. W. Anal. Chem. 2006, 78, 3326-3334.). PMID- 16689533 TI - Hybridization behavior of mixed DNA/alkylthiol monolayers on gold: characterization by surface plasmon resonance and 32P radiometric assay. AB - Nucleic acid assay from a complex biological milieu is attractive but currently difficult and far from routine. In this study, DNA hybridization from serum dilutions into mixed DNA/mercaptoundecanol (MCU) adlayers on gold was monitored by surface plasmon resonance (SPR). Immobilized DNA probe and hybridized target densities on these surfaces were quantified using 32P-radiometric assays as a function of MCU diluent exposure. SPR surface capture results correlated with radiometric analysis for hybridization performance, demonstrating a maximum DNA hybridization on DNA/MCU mixed adlayers. The maximum target surface capture produced by MCU addition to the DNA probe layer correlates with structural and conformational data on identical mixed DNA/MCU adlayers on gold derived from XPS, NEXAFS, and fluorescence intensity measurements reported in a related study (Lee, C.-Y.; Gong, P.; Harbers, G. M.; Grainger, D. W.; Castner, D. G.; Gamble, L. J. Anal. Chem. 2006, 78, 3316-3325.). MCU addition into the DNA adlayer on gold also improved surface resistance to both nonspecific DNA and serum protein adsorption. Target DNA hybridization from serum dilutions was monitored with SPR on the optimally mixed DNA/MCU adlayers. Both hybridization kinetics and efficiency were strongly affected by nonspecific protein adsorption from a complex milieu even at a minimal serum concentration (e.g., 1%). No target hybridization was detected in SPR assays from serum concentrations above 30%, indicating nonspecific protein adsorption interference of DNA capture and hybridization from complex milieu. Removal of nonsignal proteins from nucleic acid targets prior to assay represents a significant issue for direct sample-to-assay nucleic acid diagnostics from food, blood, tissue, PCR mixtures, and many other biologically complex sample formats. PMID- 16689534 TI - Isolation and characterization of glycosylphosphatidylinositol-anchored peptides by hydrophilic interaction chromatography and MALDI tandem mass spectrometry. AB - Glycosylphosphatidylinositol-anchored proteins (GPI-APs) are posttranslationally processed proteins that become tethered to the extracellular leaflet of the plasma membrane via a C-terminal glycan-like moiety. Since the first GPI-AP was described in the 1970s, more than 500 GPI-APs have been reported in a range of species, including plants, microbes, and mammals. GPI-APs are probably involved in cell signaling, cell recognition, and cell remodeling processes, and they may potentially serve as cell surface antigens or vaccine targets in pathogenic microorganisms or transformed mammalian cells. Due to the structural complexity and physicochemical properties of GPI-APs, their identification and structural characterization is a demanding analytical task. Here, we report a simple, fast and sensitive method for isolation and structural analysis of GPI-anchors using a combination of hydrophilic interaction liquid chromatography and matrix-assisted laser desorption/ionization (MALDI) quadrupole time-of-flight tandem mass spectrometry. This method allowed analysis of GPI peptides derived from low picomole levels of the porcine kidney membrane dipeptidase. Furthermore, it allowed unambiguous assignment of the omega site via amino acid sequencing of the modified peptides. GPI-anchor-specific diagnostic ions were observed by MALDI MS/MS at m/z 162, 286, 422, and 447, corresponding to glucosamine, mannose ethanolamine phosphate, glucosamine inositol phosphate, and mannose ethanolamine phosphate glucosamine, respectively. Thus, the methodology described herein may enable sensitive and specific detection of GPI-anchored peptides in large-scale proteomic studies of plasma membrane proteins. PMID- 16689535 TI - Determination of salsolinol and related catecholamines through on-line preconcentration and liquid chromatography/atmospheric pressure photoionization mass spectrometry. AB - A new analytical approach has been developed for simultaneous measurements of endogenous salsolinol and major catecholamines in brain tissue of experimental animals. This procedure involves a combination of on-line phenyl boronate affinity preconcentration and microcolumn liquid chromatography, followed by mass spectrometry equipped with an atmospheric pressure photoionization (APPI) source. Flow conditions of the APPI source were optimized for detection sensitivity while different dopants were evaluated. The on-line preconcentration was found essential for the sensitivity requirements of salsolinol measurements in the brain tissue from alcohol-preferring rats subjected to different levels of alcohol exposure. PMID- 16689536 TI - Capillarity induced solvent-actuated bonding of polymeric microfluidic devices. AB - Rapid, robust, and economical fabrication of fluidic microchannels is of fundamental importance for the successful development of disposable lab-on-a-chip devices. In this work, we present a solvent-actuated bonding method for fabricating polymeric microfluidic devices at room temperature. A PMMA sheet with an imprinted microchannel was clamped to a blank PMMA sheet, and then 80 +/- 5 muL of acetone (bonding solvent) was introduced at one end of the fluidic channel and aspirated out at the other end. As the solvent moved down the channel, capillary forces drew a fraction of the solvent into the interstitial space between the two polymeric substrates. After aspiration, the assembly was incubated in the clamp for 5 min for effective bond formation. The quantity of the bonding solvent, its water content and flow rate, along with residence time in the channel were found to have significant impact on the bond quality and the channel integrity. Microfluidic electrophoretic separations of a 400-base DNA ladder were performed in devices fabricated using this method in less than 8 min with efficiencies routinely between 2 x 10(6) and 3 x 10(6) plates/m. The simplicity and economy of this technique make it amenable for automation and mass production, which could make polymeric substrates more attractive for single-use chemical analysis devices. PMID- 16689537 TI - Chemotaxis assays of mouse sperm on microfluidic devices. AB - Sperm chemotaxis is an area of significant interest to scientists involved in reproductive science. Understanding how and when sperm cells are attracted to the egg could have profound effects on reproduction and contraception. In an effort to systematically study this problem, we have fabricated and evaluated a microfluidic device to measure sperm chemotaxis. The device was designed with a flow-through configuration using a spatially and temporally stable chemical gradient. Mouse sperm cells were introduced into the chemotaxis chamber between confluent flows of mouse ovary extract and buffer. The sperm experiencing chemotaxis swam toward the extract and were counted relative to those that swam toward the buffer. The ovary extracts were diluted from 10(2) to 10(7) times, and each extract dilution was screened for chemotaxis. Four out of six ovaries showed a strong chemotactic response at extract dilutions of 10(-3) to 10(-5). This device provided a convenient, disposable platform on which to conduct chemotaxis assays, and the flow-through design overcomes difficulties associated with distinguishing chemotaxis from trapping. PMID- 16689538 TI - Cell for simultaneous synchrotron radiation X-ray and electrochemical corrosion measurements on cultural heritage metals and other materials. AB - We describe the construction of an electrochemical cell of the Bragg type suitable for in situ synchrotron X-ray measurements on rough, heterogeneous metals such as cultural heritage alloys and simulants with corroding or passivated surfaces. The cell features a working electrode, which may be moved under remote control from a position close to an X-ray window to full immersion in the electrolyte. A pocket of electrolyte in contact with the bulk can be maintained on the working electrode surface at all times. Its thickness (typically 100-200 microm) can be controlled by adjusting the working electrode position and, independently, altering the conformation of the X-ray window with hydrostatic pressure. Alternatively, the electrode may be lowered into the bulk of the electrolyte. Early results from the cell showing a time-resolved study of the reduction of nantokite to cuprite in sodium sesquicarbonate, accompanied by corrosion potential measurements obtained in parallel, are presented here. PMID- 16689539 TI - Evaluation of hydrogel-coated glutamate microsensors. AB - Glutamate microsensors form a promising analytical tool for monitoring neuronally derived glutamate directly in the brain. However, when a microsensor is implanted in brain tissue, many factors can diminish its performance. Consequently, a thorough characterization and evaluation of a microsensor is required concerning all factors that may possibly be encountered in vivo. The present report deals with the validation of a hydrogel-coated glutamate microsensor. This microsensor is constructed by coating a carbon fiber electrode (10-microm diameter; 300-500 microm long) with a five-component redox hydrogel, in which L-glutamate oxidase, horseradish peroxidase, and ascorbate oxidase are wired via poly(ethylene glycol) diglycidyl ether to an osmium-containing redox polymer. A thin Nafion coating completes the construction. Although this microsensor was previously used in vivo, information concerning its validation is limited. In the present study, attention was given to its selectivity, specificity, calibration, oxygen dependency, biofouling, operating potential dependency, and linear range. In addition, successful microsensor experiments in microdialysate, in vitro (in organotypic hippocampal slice cultures), and in vivo (in anesthesized rats) are shown. PMID- 16689540 TI - Detection of alkaline phosphatase using surface-enhanced Raman spectroscopy. AB - A new approach was developed to detect the activity of alkaline phosphatase (ALP) enzyme at ultralow concentrations using a surface-enhanced Raman scattering (SERS) technique. The approach is based on the use of gold nanoparticles as a SERS material whereas 5-bromo-4-chloro-3-indolyl phosphate (BCIP) is used as a substrate of ALP. The enzymatic hydrolysis of BCIP led to the formation of indigo dye derivatives, which were found to be highly SERS active. For the first time, we were able to detect ALP at a concentration of approximately 4 x 10(-15) M or at single-molecule levels when ALP was incubated with BCIP for 1 h in the Tris HCl buffer. The same technique also was successfully employed to detect surface immobilized avidin, and a detection limit of 10 ng/mL was achieved. This new technique allows the detection of both free and labeled ALP as a Raman probe in enzyme immunoassays, immunoblotting, and DNA hybridization assays at ultralow concentrations. PMID- 16689541 TI - Automatic deconvolution of isotope-resolved mass spectra using variable selection and quantized peptide mass distribution. AB - We present an algorithm for the deconvolution of isotope-resolved mass spectra of complex peptide mixtures where peaks and isotope series often overlap. The algorithm formulates the problem of mass spectrum deconvolution as a classical statistical problem of variable selection, which aims to interpret the spectrum with the least number of peptides. The LASSO method is used to perform automatic variable selection. The algorithm also makes use of the quantized distribution of peptide masses in the NCBInr database after in silico trypsin digestion as filters to aid the deconvolution process. Errors in the expected isotope pattern are accounted for to avoid spurious isotope series. The effectiveness of the algorithm is demonstrated with annotated ESI spectrum of known peptides for which the peaks and isotope series are highly overlapping. The algorithm successfully finds all correct masses in the experimental spectrum, except for one spectrum where an additional refinement procedure is required to obtain the correct results. Our results compare favorably to those from a widely used commercial program. PMID- 16689542 TI - Development and evaluation of a candidate reference measurement procedure for the determination of 19-norandrosterone in human urine using isotope-dilution liquid chromatography/tandem mass spectrometry. AB - 19-Norandrosterone (19-NA) is the major metabolite of the steroid nandrolone, one of the most commonly abused anabolic androgenic agents. 19-NA exists mainly as the glucuronide form in human urine. A candidate reference measurement procedure for 19-NA in urine involving isotope dilution coupled with liquid chromatography/tandem mass spectrometry (LC/MS/MS) has been developed and critically evaluated. The 19-NA glucuronide was enzymatically hydrolyzed, and the 19-NA along with its internal standard (deuterated 19-NA) was extracted from urine using liquid-liquid extraction prior to reversed-phase LC/MS/MS. The accuracy of the measurement of 19-NA was evaluated by a recovery study of added 19-NA. The recovery of the added 19-NA ranged from 99.1 to 101.4%. This method was applied to the determination of 19-NA in urine samples fortified with 19-NA glucuronide at three different concentrations (equivalent to 1, 2, and 10 ng/mL 19-NA). Excellent reproducibility was obtained with within-set coefficients of variation (CVs) ranging from 0.2 to 1.2%, and between-set CVs ranging from 0.1 to 0.5%. Excellent linearity was also obtained with correlation coefficients of all linear regression lines (measured intensity ratios vs mass ratios) ranging from 0.9997 to 0.9999. The detection limit for 19-NA at a signal-to-noise ratio of approximately 3 was 16 pg. The mean results of 19-NA yielded from hydrolysis of 19-NA glucuronide compared well with the theoretical values (calculated from the conversion of 19-NA glucuronide to 19-NA) with absolute relative differences ranging from 0.2 to 1.4%. This candidate reference measurement procedure for 19 NA in urine, which demonstrates good accuracy and precision and low susceptibility to interferences, can be used to provide an accuracy base to which routine methods for 19-NA can be compared and that will serve as a standard of higher order for measurement traceability. PMID- 16689543 TI - Construction of a biomimetic surface on microfluidic chips for biofouling resistance. AB - A biomimetic surface has been formed on the poly(methyl methacrylate) (PMMA) microfluidic chips for biofouling resistance on the basis of a simple modification. Accordingly, an amphiphilic phospholipid copolymer of 2 methacryloyloxyethyl phosphorylcholine and n-butyl methacrylate (PMB) was developed to introduce the phosphorylcholine functional groups onto the PMMA surface via the anchoring of hydrophobic n-butyl methacrylate units. The 2 methacryloyloxyethyl phosphorylcholine segments could form hydrophilic domains, considered to be located on the surface, to provide a biocompatible surface. X ray photoelectron spectroscopy and Fourier transform infrared spectra confirmed the success of surface functionalization. The PMB-modified microchips containing phosphorylcholine moieties exhibited more stable electroosmotic mobility compared with the untreated one. In addition to being characterized for minimized nonspecific adhesion of serum proteins and plasma platelets, the PMB functionalized microchannels have been exemplified by electrophoresis of proteins. This one-step procedure offers an effective approach for a biomimetic surface design on microfluidic chips, which is promising in high-throughput and complex biological analysis. PMID- 16689544 TI - Peak capacity optimization of peptide separations in reversed-phase gradient elution chromatography: fixed column format. AB - The optimization of peak capacity in gradient elution RPLC is essential for the separation of multicomponent samples such as those encountered in proteomic research. In this work, we study the effect of gradient time (tG), flow rate (F), temperature (T), and final eluent strength (phi(final)) on the peak capacity of separations of peptides that are representative of the range in peptides found in a tryptic digest. We find that there are very strong interactions between the individual variables (e.g., flow rate and gradient time) which make the optimization quite complicated. On a given column, one should first set the gradient time to the longest tolerable and then set the temperature to the highest achievable with the instrument. Next, the flow rate should be optimized using a reasonable but arbitrary value of phi(final). Last, the final eluent strength should be adjusted so that the last solute elutes as close as possible to the gradient time. We also develop an easily implemented, highly efficient, and effective Monte Carlo search strategy to simultaneously optimize all the variables. We find that gradient steepness is an important parameter that influences peak capacity and an optimum range of gradient steepness exists in which the peak capacity is maximized. PMID- 16689545 TI - Mass defect labeling of cysteine for improving peptide assignment in shotgun proteomic analyses. AB - A method for improving the identification of peptides in a shotgun proteome analysis using accurate mass measurement has been developed. The improvement is based upon the derivatization of cysteine residues with a novel reagent, 2,4 dibromo-(2'-iodo)acetanilide. The derivitization changes the mass defect of cysteine-containing proteolytic peptides in a manner that increases their identification specificity. Peptide masses were measured using matrix-assisted laser desorption/ionization Fourier transform ion cyclotron mass spectrometry. Reactions with protein standards show that the derivatization of cysteine is rapid and quantitative, and the data suggest that the derivatized peptides are more easily ionized or detected than unlabeled cysteine-containing peptides. The reagent was tested on a 15N-metabolically labeled proteome from M. maripaludis. Proteins were identified by their accurate mass values and from their nitrogen stoichiometry. A total of 47% of the labeled peptides are identified versus 27% for the unlabeled peptides. This procedure permits the identification of proteins from the M. maripaludis proteome that are not usually observed by the standard protocol and shows that better protein coverage is obtained with this methodology. PMID- 16689546 TI - Stereoisomeric separation of flavanones and flavanone-7-O-glycosides by capillary electrophoresis and determination of interconversion barriers. AB - The stereoisomeric separation of several flavanones and flavanone-7-O-glycosides has been achieved with capillary electrophoresis by adding native cyclodextrins or cyclodextrin derivatives to the background electrolyte. As an alternative method, micellar electrokinetic chromatography with sodium cholate as a chiral surfactant has been used for the epimeric separation of two flavanone-7-O glycosides. The effect of buffer systems containing mixtures of cyclodextrin with either sodium dodecyl sulfate or sodium cholate upon the chiral recognition of flavanones and flavanone-7-O-glycosides as well as the variation of the background electrolyte (concentration of buffer and surfactant, pH value, organic modifier), and its influence on the resolution factor Rs was investigated. Temperature- and pH-dependent enantiomerization or epimerization barriers of several flavanones (naringenin, homoeriodictyol) and flavanone-7-O-glycosides (naringin, neohesperidin, prunin, narirutin) in basic media (pH values of 9-11) have been observed. Interconversion profiles featuring characteristic plateau formation of the elution pattern were observed at high pH and evaluated with the simulation software ChromWin to determine rate constants k(T) and Eyring activation parameters, DeltaG#(T), DeltaH#, and DeltaS#. PMID- 16689547 TI - Profiling of hydrophilic proteins from Olea europaea olive pollen by MALDI TOF mass spectrometry. AB - The antigenic profile of Olea europaea pollen from different Mediterranean cultivars was obtained by MALDI mass spectrometry. A simple procedure of chemical fractionation of the whole antigen extract was developed, whereby less complex, or pure, fractions of antigen candidate were obtained prior to mass spectrometric analysis. Some of the features of protein structure and distribution probably depend on cultivar adaptation to the environment. The profilings of pollen proteins thus obtained allow the distinction of the analyzed cultivars into three distinct groups: (i) those characterized by a low Ole e 1 content; (ii) those over-enriched in Ole e 1 and (iii) that containing Ole e 3 and Ole e 7 only. The latter consists of at least four isoforms differing by the degree of glycosilation. These results demonstrate that the proposed experimental procedure, can supply valuable information on the antigens' micro heterogeneity. PMID- 16689548 TI - Electron multiplying charge-coupled device camera based fluorescence correlation spectroscopy. AB - A fluorescence correlation spectroscopy (FCS) setup is built with an electron multiplying charge-coupled device camera. Although the instrument has a limited time resolution of 4 ms, compared to 0.1-0.2 mus for common instruments using avalanche photodiodes, it allows multiplexing of FCS measurements, has a software adjustable pinhole after data collection, performs flow speed as well as flow direction measurements in microchannels and could be used to do spectral FCS. Measurements are performed on fluorescent dyes and polystyrene beads in high viscosity media and on epidermal growth factor receptors in Chinese hamster ovary cells. Using real measurements on single spots, multiplexing of focal spots and detection elements are simulated and the results are discussed. PMID- 16689549 TI - Carbon-13 labeled polymers: an alternative tracer for depth profiling of polymer films and multilayers using secondary ion mass spectrometry. AB - 13C labeling is introduced as a tracer for depth profiling of polymer films and multilayers using secondary ion mass spectrometry (SIMS). Deuterium substitution has traditionally been used in depth profiling of polymers but can affect the phase behavior of the polymer constituents with reported changes in both bulk phase behavior and surface and interfacial interactions. SIMS can provide contrast by examining various functional groups, chemical moieties, or isotopic labels. 13C-Labeled PS (13C-PS) and unlabeled PS (12C-PS) and PMMA were synthesized using atom-transfer radical polymerization and assembled in several model thin-film systems. Depth profiles were recorded using a Cameca IMS-6f magnetic sector mass spectrometer using both 6.0-keV impact energy Cs+ and 5.5 keV impact energy O2+ primary ion bombardment with detection of negative and positive secondary ions, respectively. Although complete separation of 12C1H from 13C is achieved using both primary ion species, 6.0-keV Cs+ clearly shows improved detection sensitivity and signal-to-noise ratio for detection of 12C, 12C1H, and 13C secondary ions. The use of Cs+ primary ion bombardment results in somewhat anomalous, nonmonotonic changes in the 12C, 12C1H, and 13C secondary ion yields through the PS/PMMA interface; however, it is shown that this behavior is not due to sample charging. Through normalization of the 13C secondary ion yield to the total C (12C + 13C) ion yield, the observed effects through the PS/PMMA interface can be greatly minimized, thereby significantly improving analysis of polymer films and multilayers using SIMS. Mass spectra of 13C-PS and 12C-PS were also analyzed using a PHI TRIFT I time-of-flight mass spectrometer, with 15-keV Ga+ primary ion bombardment and detection of positive secondary ions. The (12)C7(1)H7 ion fragment and its 13C-enriched analogues have significant secondary ion yields with negligible mass interferences, providing an early indication of the potential for future use of this technique for cluster probe depth profiling of high molecular weight 13C-labeled fragments. PMID- 16689550 TI - Analysis of energy-resolved mass spectra at MSn in a pursuit to characterize structural isomers of oligosaccharides. AB - Analyses of energy-resolved mass spectra (ERMS) of structural isomeric oligosaccharides were carried out under collision-induced dissociation conditions at different stages of MSn. Quantitative analyses of energy-resolved curves revealed that these curves can be used to distinguish anomeric configurations and such analysis can be made regardless of the stages of MSn. Furthermore, parameters obtained from the sigmoidal shaped curves obtained for ERMS were found to be used as the structural descriptors after statistical analysis. An important finding is that analyses of these descriptors depend on neither the m/z values nor the height factors. PMID- 16689551 TI - Electrochemical oxidation of oxalic acid at highly boron-doped diamond electrodes. AB - Electrochemical oxidation of oxalic acid has been investigated at bare, highly boron-doped diamond electrodes. Cyclic voltammetry and flow injection analysis with amperometric detection were used to study the electrochemical reaction. Hydrogen-terminated diamonds exhibited well-defined peaks of oxalic acid oxidation in a wide pH range. A good linear response was observed for a concentration range from 50 nM to 10 microM, with an estimated detection limit of approximately 0.5 nM (S/N = 3). In contrast, oxygen-terminated diamonds showed no response for oxalic acid oxidation inside the potential window, indicating that surface termination contributed highly to the control of the oxidation reaction. An investigation with glassy carbon electrodes was conducted to confirm the surface termination effect on oxalic acid oxidation. Although a hydrogen terminated glassy carbon electrode showed an enhancement of signal-to-background ratio in comparison with untreated glassy carbon, less stability of the current responses was observed than that at hydrogen-terminated diamond. PMID- 16689552 TI - Universal microfluidic gradient generator. AB - The study of cellular responses to chemical gradients in vitro would greatly benefit from experimental systems that can generate precise and stable gradients comparable to chemical nonhomogeneities occurring in vivo. Recently, microfluidic devices have been demonstrated for linear gradient generation for biological applications with unmatched accuracy and stability. However, no systematic approach exists at this time for generating other gradients of target spatial configuration. Here we demonstrate experimentally and provide mathematical proof for a systematic approach to generating stable gradients of any profile by the controlled mixing of two starting solutions. PMID- 16689553 TI - Ecopharmacology: a new topic of importance in pharmacovigilance. PMID- 16689554 TI - Data mining in the US using the Vaccine Adverse Event Reporting System. AB - The US Vaccine Adverse Event Reporting System (VAERS), which is charged with vigilance for detecting vaccine-related safety issues, faces an increasingly complex immunisation environment. Since 1990, steady increases in vaccine licensing and distribution have resulted in increasing numbers of reports to VAERS. Prominent features of current reports include more routine vaccine co administration and frequent reports of new postvaccination clinical syndromes. Data-mining methods, based on disproportionality analyses, are one strategy being pursued by VAERS researchers to increase the utility of its complex database. The types of analyses used include proportional reporting ratios, association rule discovery, and various 'historic limits' methods that compare observed versus expected event counts. The use of such strategies in VAERS has been primarily supplemental and retrospective. Signals for inactivated influenza, typhoid and tetanus toxoid-containing vaccines have been successfully identified. Concerns flagged through data mining should always be subject to clinical case review as a first evaluation step. Persistent issues should be subject to formal hypothesis testing in large linked databases or other controlled-study settings. Automated data-mining techniques for prospective use are currently undergoing development and evaluation within VAERS. Their use (as one signal-detection tool among many) by trained medical evaluators who are aware of system limitations is one legitimate approach to improving the ability of VAERS to generate vaccine-safety hypotheses. Such approaches are needed as more new vaccines continue to be licensed. PMID- 16689555 TI - Under-reporting of adverse drug reactions : a systematic review. AB - The purpose of this review was to estimate the extent of under-reporting of adverse drug reactions (ADRs) to spontaneous reporting systems and to investigate whether there are differences between different types of ADRs. A systematic literature search was carried out to identify studies providing a numerical estimate of under-reporting. Studies were included regardless of the methodology used or the setting, e.g. hospital versus general practice. Estimates of under reporting were either extracted directly from the published study or calculated from the study data. These were expressed as the percentage of ADRs detected from intensive data collection that were not reported to the relevant local, regional or national spontaneous reporting systems. The median under-reporting rate was calculated across all studies and within subcategories of studies using different methods or settings. In total, 37 studies using a wide variety of surveillance methods were identified from 12 countries. These generated 43 numerical estimates of under-reporting. The median under-reporting rate across the 37 studies was 94% (interquartile range 82-98%). There was no significant difference in the median under-reporting rates calculated for general practice and hospital-based studies. Five of the ten general practice studies provided evidence of a higher median under-reporting rate for all ADRs compared with more serious or severe ADRs (95% and 80%, respectively). In comparison, for five of the eight hospital-based studies the median under-reporting rate for more serious or severe ADRs remained high (95%). The median under-reporting rate was lower for 19 studies investigating specific serious/severe ADR-drug combinations but was still high at 85%. This systematic review provides evidence of significant and widespread under reporting of ADRs to spontaneous reporting systems including serious or severe ADRs. Further work is required to assess the impact of under-reporting on public health decisions and the effects of initiatives to improve reporting such as internet reporting, pharmacist/nurse reporting and direct patient reporting as well as improved education and training of healthcare professionals. PMID- 16689557 TI - Overcoming 'ageism' bias in the treatment of hypercholesterolaemia : a review of safety issues with statins in the elderly. AB - Atherosclerosis is a progressive, lifelong condition that is the leading cause of death among middle-aged and elderly individuals aged > or =65 years. Up to 80% of elderly patients are found to have evidence of obstructive coronary heart disease at autopsy. Demographic trends, including the advancing median age and life expectancy of Western societies, suggest that a large share of the burden of atherosclerotic plaque is likely to be borne by elderly individuals. These trends are in part due to increases in a number of chronic diseases associated with adverse cardiovascular outcomes, including metabolic syndrome, diabetes mellitus and chronic kidney disease. Because the elderly have a higher attributable risk of coronary heart disease as a result of hypercholesterolaemia, more coronary deaths and overall events can be prevented via treatment in this age group compared with younger persons with hypercholesterolaemia. The efficacy, safety and tolerability of HMG-CoA reductase inhibitors (statins) have been confirmed in randomised, controlled, multicentre trials involving large numbers of patients aged > or =65 years. Although muscle symptoms such as myalgia are relatively common adverse events, more severe signs of myolysis such as myopathy and rhabdomyolysis are rare, but their risk is elevated by conditions (e.g. concomitant medications) that increase the systemic exposure of these agents. Statins differ in their susceptibility to increases in systemic exposure, but most statins have been demonstrated to be well tolerated and safe when administered to elderly patients. These favourable clinical findings should help clinicians counter highly prevalent 'ageism' bias in statin prescribing, whereby elderly patients, particularly those at highest cardiovascular risk, are often denied the benefits of statins without any meaningful foundation. PMID- 16689556 TI - Effects of maternally administered drugs on the fetal and neonatal kidney. AB - The number of pregnant women and women of childbearing age who are receiving drugs is increasing. A variety of drugs are prescribed for either complications of pregnancy or maternal diseases that existed prior to the pregnancy. Such drugs cross the placental barrier, enter the fetal circulation and potentially alter fetal development, particularly the development of the kidneys. Increased incidences of intrauterine growth retardation and adverse renal effects have been reported. The fetus and the newborn infant may thus experience renal failure, varying from transient oligohydramnios to severe neonatal renal insufficiency leading to death. Such adverse effects may particularly occur when fetuses are exposed to NSAIDs, ACE inhibitors and specific angiotensin II receptor type 1 antagonists. In addition to functional adverse effects, in utero exposure to drugs may affect renal structure itself and produce renal congenital abnormalities, including cystic dysplasia, tubular dysgenesis, ischaemic damage and a reduced nephron number. Experimental studies raise the question of potential long-term adverse effects, including renal dysfunction and arterial hypertension in adulthood. Although neonatal data for many drugs are reassuring, such findings stress the importance of long-term follow-up of infants exposed in utero to certain drugs that have been administered to the mother. PMID- 16689559 TI - An experimental value for the B(1u) C-H stretch mode in benzene. AB - We here present experimental infrared spectra on two (C(6)H(6))(C(6)D(6)) benzene dimer isomers in the gas phase. The spectra show that the two benzene molecules in the dimer are symmetrically inequivalent and have distinct IR signatures. One of the two molecules is in a site of low symmetry, which leads to the IR activation of fundamental modes that are IR forbidden by symmetry in the monomer. In the spectra, all four fundamental C-H stretch modes of benzene are observed. Modes in the dimer are shifted up to 3 cm(-1) to the red, compared to the modes that are known for the monomer. For the nu(13) B(1u) C-H stretch fundamental mode of benzene, a first experimental value of 3015(+2) (-5) cm(-1) is determined, in excellent agreement with anharmonic frequency calculations presented here. PMID- 16689558 TI - Adverse reactions induced by NSAIDs and antibacterials: analysis of spontaneous reports from the Sicilian regional database. AB - OBJECTIVES: To (i) evaluate the suspected adverse drug reactions (ADRs) related to NSAIDs and antibacterials that were reported to Sicilian local health officers by healthcare professionals; and (ii) to detect new or serious potential signals of alarm related to these two widely used drug categories. METHODS: We selected all the spontaneous reports of ADRs sent between January 1998 and June 2004 and analysed those attributed to NSAIDs and systemic antibacterials, applying proportional reporting ratio (PRR) methodology. PRRs >2, chi(2) >4 and >3 ADRs were regarded as signals. RESULTS: During the period considered, 1585 reports of ADRs were received overall (42.6% serious), with an annual reporting rate of approximately 49.1 reports per million inhabitants on average; 351 referred to systemic antibacterials, and 179 to NSAIDs. There were 174 (49.6%) reports of serious ADRs associated with antimicrobials and 108 (60.3%) associated with NSAIDs. Disproportionality was observed, in particular for anaphylactic shock induced by ceftriaxone (all reports were associated with off-label use of the drug), photosensitivity reaction induced by lomefloxacin (administered in the summer), hepatitis induced by nimesulide (three cases leading to liver transplantation) and vasculitis induced by nimesulide. CONCLUSION: Our analysis highlighted several signals of alarm deserving further investigation or measures to influence prescribing. This study underlines the value of a regional centre in identifying local factors (such as prescribing patterns) that may increase the prevalence of serious ADRs. PMID- 16689560 TI - Transient behavior at the nanoscale. AB - Transient and steady state responses of a system to an input are well-known features of materials and systems in science and engineering. These responses depend on the intrinsic parameters of the system and on the nature of the input. We find that a system comprised of nanosized features no longer shows the typical stationary characteristics as their microscopic or solid-state counterparts. Interestingly, because of the chemistry of the nanostructure, thermal motion of the atoms, and external fields, the nanosized system shows extended electrical transient behavior, compatible with highly nonlinear features such a negative differential resistance and hysteresis. PMID- 16689561 TI - Reduced multireference coupled cluster method with singles and doubles: Perturbative corrections for triples. AB - The reduced multireference coupled-cluster method with singles and doubles (RMR CCSD) that employs multireference configuration interaction wave function as an external source for a small subset of approximate connected triples and quadruples, is perturbatively corrected for the remaining triples along the same lines as in the standard CCSD(T) method. The performance of the resulting RMR CCSD(T) method is tested on four molecular systems, namely, the HF and F(2) molecules, the NO radical, and the F(2) (+) cation, representing distinct types of molecular structure, using up to and including a cc-pVQZ basis set. The results are compared with those obtained with the standard CCSD(T), UCCSD(T), CCSD(2), and CR CCSD(T) methods, wherever applicable or available. An emphasis is made on the quality of the computed potentials in a broad range of internuclear separations and on the computed equilibrium spectroscopic properties, in particular, harmonic frequencies omega(e). It is shown that RMR CCSD(T) outperforms other triply corrected methods and is widely applicable. PMID- 16689562 TI - A general and efficient pseudopotential Fourier filtering scheme for real space methods using mask functions. AB - A scheme for the Fourier filtering of pseudopotentials in real space calculations is proposed, in order to reduce the artifact of positional energy dependence ("egg box" effect). It is based on an improved version of the mask function method poposed by Wang [Phys. Rev. B 64, 201107/1 (2001)]. It is easy to implement, efficient, and accurate. By using atom-centered compensation charges, the local part of the pseudopotential becomes short ranged and can be filtered on the same footing as the nonlocal parts. A major advantage of the approach is that a generic set of parameters can be used for different pseudopotentials. A balanced parameter set is derived and validated. In this context a strategy to monitor the extent of grid dependence is introduced. It is found that, given a sufficiently fine grid spacing is used to represent the atomic valence density, the positional energy dependence can be reduced below 0.1 mhartree for all investigated atoms. On the example of a D(3h) symmetric Si(5) cluster and the C(60) molecule it is demonstrated that the artificial symmetry breaking of both bond lengths and orbital energies can substantially be reduced by the filtering scheme. PMID- 16689563 TI - Time-dependent density functional theory with the generalized restricted unrestricted approach. AB - This work presents the derivation, implementation, and first applications of the generalized restricted-unrestricted method based on the density functional Kohn Sham formalism. By using a spin-restricted Kohn-Sham representation for the reference state the well-known spin contamination problem is avoided, while the unrestricted representation of the perturbation response retains a proper description of spin polarization. The formulation is a generalization of our previous implementation of the restricted-unrestricted method [Z. Rinkevicius et al., J. Chem. Phys. 121, 7614 (2004)], as it accounts for the full unrestricted response instead of describing the spin polarization in terms of triplet operators only. The purpose of this paper is to investigate the role of the generalization employed and demonstrate its numerical performance. For this purpose we focus on isotropic hyperfine coupling constants of a set of organic radicals and transition metal compounds. For both classes of molecules we observe that the effect of neglecting singlet excitation operators in the response part of restricted-unrestricted formalism changes the calculated hyperfine coupling parameters by a few MHz. The obtained results confirm the validity of the approximation used in the simplified restricted-unrestricted approach, i.e. spin polarization can, in most cases, be adequately described by an account of only triplet operators in the response term. PMID- 16689564 TI - A post-Hartree-Fock model of intermolecular interactions: inclusion of higher order corrections. AB - We have previously demonstrated that the dipole moment of the exchange hole can be used to derive intermolecular C(6) dispersion coefficients [J. Chem. Phys. 122, 154104 (2005)]. This was subsequently the basis for a novel post-Hartree Fock model of intermolecular interactions [J. Chem. Phys. 123, 024101 (2005)]. In the present work, the model is extended to include higher-order dispersion coefficients C(8) and C(10). The extended model performs very well for prediction of intermonomer separations and binding energies of 45 van der Waals complexes. In particular, it performs twice as well as basis-set extrapolated MP2 theory for dispersion-bound complexes, with minimal computational cost. PMID- 16689565 TI - A theoretical study of the chiroptical properties of molecules with isotopically engendered chirality. AB - There has been a considerable interest in the chiroptical properties of molecules whose chirality is exclusively due to an isotopic substitution and numerous examples for the electronic circular dichroism (CD) spectra of isotopically chiral systems have been reported in literature. Four different explanations have been proposed for the mechanism as to how the isotopic substitution induces a chiral perturbation of the otherwise achiral electronic wave function; however, up to now no conclusive answer has been given about the dominating effect responsible for the experimental observations. In this study we will present, for the first time, fully quantum-mechanical calculations of the CD spectra of three different molecular systems with isotopically engendered chirality. As examples, we consider the spectra of organic molecules with ketone and alpha-diketone carbonyl and diene chromophores. The effect of vibronic couplings for the reorientation of the electric and magnetic transition dipole moments is taken into account within the Herzberg-Teller approximation. The ground and excited state geometries and vibrational normal modes are obtained with (time-dependent) density functional theory [(TD)DFT], while the vibronic coupling effects are calculated at the TDDFT and density functional theory/multireference configuration interaction (DFT/MRCI) levels of theory. Generally, the band shapes of the experimental CD spectra are reproduced very well, and also the absolute CD intensities from the simulations are of the right order of magnitude. The sign and the intensity of the CD band are determined by a delicate balance of the contributions of a large number of individual vibronic transitions, and it is found that the vibrational normal modes with a large displacement are dominant. The separation of the calculated CD spectrum into the different contributions due to the overlap of the in-plane and out-of-plane components (regarding the symmetry plane of the unsubstituted molecule) of the electric and magnetic transition dipole moments yields information about the influence of the vibronic coupling effects for the reorientation of the corresponding transition dipole moments. In conclusion, the calculations clearly show that vibronic effects are responsible or at least dominant for the chiroptical properties of isotopically chiral organic molecules. PMID- 16689566 TI - Determination of the CO3- bond strength via the resonant two-photon photodissociation threshold: Electronic and vibrational spectroscopy of CO3 .Ar(n). AB - We use a two-laser pump-probe technique coupled with messenger atom tagging to determine the bond energy of O(-) to CO(2) in the CO(3) (-) ion, a prevalent species in the upper atmosphere. In this technique, the argon-tagged ion is first electronically excited using a visible laser, then irradiated with a tunable near infrared beam across the CO(2)...O(-) dissociation threshold while O(-) products are monitored. This method yields a bond energy of 2.79+/-0.05 eV, which is about 0.5 eV higher than previously reported. Combining this with the well-known heats of formation of O(-) and CO(2), 105.6 and -393.1 kJmol, respectively [Thermodynamic Properties of Individual Substances, edited by L. V. Gurvich, I. V. Veyts, and C. B. Alcock (Hemisphere, New York, 1989), Vol. 1 and CODATA Thermodynamic Tables, edited by O. Garvin, V. B. Parker, and J. H. J. White (Hemisphere, New York, 1987)], yields the CO(3) (-) heat of formation: DeltaH(0) (0)=-556.7+/-4.8 kJmol. The one-photon (i.e., linear) infrared and electronic spectra of CO(3) (-) are also presented and compared to those obtained previously. The one-photon electronic spectrum is nearly identical to two-photon spectra, implying that argon does not significantly perturb the ion or its symmetry. The infrared spectrum is drastically different than that obtained in an argon matrix, however, indicating that the ion is likely distorted in the matrix environment. PMID- 16689567 TI - Quantum real wave-packet dynamics of the N(4S)+NO(X2Pi)-->N2(X1Sigma(g)+)+O(3P) reaction on the ground and first excited triplet potential energy surfaces: rate constants, cross sections, and product distributions. AB - The reaction N+NO-->N(2)+O was studied by means of the time-dependent real wave packet (WP) method and the J-shifting approximation. We consider the ground 1 (3)A(") and first excited 1 (3)A(') triplet states, which correlate with both reactants and products, using analytical potential energy surfaces (PESs) recently developed in our group. This work extends our previous quantum dynamics study, and probabilities, cross sections, and rate constants were calculated and interpreted on the basis of the different shapes of the PESs (barrierless 1 (3)A(") and with barrier 1 (3)A(') surfaces, respectively). The WP rate constant (k(1)) shows a weak dependence on T(200-2500 K), as the dominant contribution to reactivity is provided by the barrierless ground PES. There is a good agreement of WP k(1) with the measurements and variational transition state theory (VTST) data, and also between the WP and VTST k(1)(1 (3)A(")) results. Nevertheless, there is a large discrepancy between the WP and VTST k(1)(1 (3)A(')) results. Product state distributions were also calculated for the much more reactive 1 (3)A(") PES. There is an excellent agreement with the experimental average fraction of vibrational energy in N(2)(25+/-3%), the only measured dynamics property of this reaction. PMID- 16689568 TI - Correlation energy extrapolation by intrinsic scaling. V. Electronic energy, atomization energy, and enthalpy of formation of water. AB - The method of correlation energy extrapolation by intrinsic scaling, recently introduced to obtain accurate molecular electronic energies, is used to calculate the total nonrelativistic electronic ground state energy of the water molecule. Accurate approximations to the full configuration interaction energies are determined for Dunning's [J. Chem. Phys. 90, 1007 (1989)] correlation-consistent double-, triple- and quadruple-zeta basis sets and then extrapolated to the complete basis set limit. The approach yields the total nonrelativistic energy 76.4390+/-0.0004 hartree, which compares very well with the value of -76.4389 hartree derived from experiment. The energy of atomization is recovered within 0.1 mh. The enthalpy of formation, which is obtained in conjunction with our previous calculation of the dissociation energy of the oxygen molecule, is recovered within 0.05 mh. PMID- 16689569 TI - Stereographic projection path-integral simulations of (HF)n clusters. AB - We perform several quantum canonical ensemble simulations of (HF)(n) clusters. The HF stretches are rigid, and the stereographic projection path-integral method is employed for the simulation in the resulting curved configuration space. We make use of the reweighted random series techniques to accelerate the convergence of the path-integral simulation with respect to the number of path coefficients. We develop and test estimators for the total energy and heat capacity based on a finite difference approach for non-Euclidean spaces. The quantum effects at temperatures below 400 K are substantial for all sizes. We observe interesting thermodynamic behaviors in the quantum simulations of the octamer and the heptamer. PMID- 16689570 TI - Conversion pathways between a fullerene and a ring among C(20) clusters by a sphere contracting walk method: Remarkable difference in local potential energy landscapes around the fullerene and the ring. AB - Conversion pathways from a fullerene to a ring and vice versa among C(20) clusters have been explored. A new technique has been developed for discovering multistep reaction pathways, which can be searched by finding constrained energy minima on spheres whose surfaces are contracting from the starting point to the terminal. Local landscapes that appeared on the pathways were found to be considerably different around the fullerene and the ring, from which one can make a new interpretation for no generation of the C(20) fullerene in laser vaporization of graphite. PMID- 16689571 TI - Coherent ultrafast photoelectron spectroscopy in Br2: an alternative method for measuring Delta v not equal 0 transitions in Rydberg-to-Rydberg excitations. AB - A two-state vibrational wave packet is prepared in a low-lying 4d[12](1 or 2) Rydberg state of jet cooled Br(2) (4d, v(')=3 and v(')=4) by two-photon excitation with 266.5 nm pulses from an ultrafast laser. The wave packet is detected by autoionization following excitation with time-delayed 800 nm pulses to the n=8 (v(+)=4) and n=9 (v(+)=3) Rydberg states in the (2)Pi(12) angular momentum core state. Autoionization of each state occurs to the (2)Pi(32) state of the ion through spin-orbit ionization. Photoelectron spectroscopy is used to differentiate between the n=8 and n=9 ejected photoelectrons. Detection of the wave packet recurrences via the n=8 and n=9 Rydberg states reveals a pi phase shift difference of the recurrences between the two final states. In each case, Delta v not equal 0 transitions are observed since wave packet recurrences are detected. By fitting the observed phase change of the recurrences with a simple model for the overlap amplitudes and assumptions about the potentials, we estimate, within the context of the model, that approximately 0.6% of the transitions may be attributed to Delta v= +/- 1 transitions between the initial Rydberg superposition state and the final Rydberg detection state. PMID- 16689572 TI - A theoretical anharmonic study of the infrared absorption spectra of FHF-, FDF-, OHF-, and ODF- anions. AB - Anharmonic vibrational frequencies, equilibrium bond lengths, rotational constants, and vibrational absorption spectra have been calculated for the triatomic anions, FHF(-) and OHF(-), and the heavier isotopomers FDF(-) and ODF( ). The triatomic anions are assumed to maintain a collinear configuration throughout all calculations, so only the symmetric (nu(1)) and asymmetric (nu(3)) stretching modes are considered. The two-dimensional permanent dipole surfaces and potential energy surfaces are then constructed along bond coordinates, using high-level ab initio methods. Fundamental and combination bands are obtained from the vibrational eigenfunctions, resulting in anharmonic frequencies, which can be compared with the available theoretical and experimental data. The agreement is very good, especially for the pure symmetric modes, while the asymmetric ones show larger discrepancies, presumably due to the neglected coupling between stretching and bending modes. Strong inverse anharmonicity is found in the level spacing of the asymmetric modes, for both FHF(-) and OHF(-) anions. The calculated mixed modes (nnu(1)+mnu(3), n, m=0-3) also agree reasonably with the few available experimental data, supporting our model. Based on the vibrational eigenfunctions, isotope effects are also rationalized. Infrared absorption spectra are calculated from the dipole autocorrelation function for FHF(-) and FDF(-), and for OHF(-) and ODF(-). Peak locations and relative intensities are assigned in terms of the fundamental and mixed transitions. PMID- 16689573 TI - High-resolution laser excitation spectroscopy of the A2E-X2A1 transition of SrCH3. AB - High-resolution laser excitation spectroscopy has been used to record the A (2)E X (2)A(1) electronic transition of SrCH(3) in a laser ablation/molecular jet source. Transitions arising from the K(')=1<--K(")=0, K(')=0<--K(")=1, and K(')=2<--K(")=1 subbands have been observed and assigned. The data were modeled with (2)E and (2)A(1) symmetric top Hamiltonian matrices in a Hund's case (a) basis, using a least squares fitting program. Rotational and fine structure parameters for the A (2)E state were determined. A comparison of the spin-orbit energy separation in the A (2)E state to other strontium containing free radicals showed that the Jahn-Teller effect is negligible. The spin-rotation constants for the A (2)E state were calculated using the pure precession model and were found to be in good agreement with the experimentally determined parameters. These calculations suggest that the A (2)E state of SrCH(3) is not entirely of p orbital character. The rotational constants were used to estimate the structural parameters of SrCH(3) in the A (2)E state. The strontium-carbon bond length was found to decrease by approximately 0.006 A, and the hydrogen-carbon-hydrogen bond angle opened by approximately 0.8 degrees compared to the X (2)A(1) state, similar to the geometry changes observed for CaCH(3). PMID- 16689574 TI - High resolution spectroscopy and potential determination of the (3)1Pi state of NaCs. AB - The (3)(1)Pi state of the NaCs molecule was studied by high resolution Fourier transform spectroscopy. The (3)(1)Pi-->X (1)Sigma(+) laser induced fluorescence was excited by an Ar(+) ion laser or by a single-mode frequency-doubled cw Nd:YAG laser. The presence of argon buffer gas yielded rich rotational relaxation spectra allowing to enlarge the data set for the (3)(1)Pi state term values, as well as to observe Lambda splittings in a wide range of vibrational (v(')) and rotational (J(')) quantum numbers. The data field includes about 820 energy levels of (3)(1)Pi NaCs in the range from v(')=0 to 37 and from J(')=3 to 190, which corresponds to ca. 95% of the potential well depth. Direct fit of the potential energy curve to the level energies is realized using the inverted perturbation approach method; a set of Dunham coefficients is also presented. PMID- 16689575 TI - [Pb(H2O)]2+ and [Pb(OH)]+: four-component density functional theory calculations, correlated scalar relativistic constrained-space orbital variation energy decompositions, and topological analysis. AB - Within the scope of studying the molecular implications of the Pb(2+) cation in environmental and polluting processes, this paper reports Hartree-Fock and density functional theory (B3LYP) four-component relativistic calculations using an all-electron basis set applied to [Pb(H(2)O)](2+) and [Pb(OH)](+), two complexes expected to be found in the terrestrial atmosphere. It is shown that full-relativistic calculations validate the use of scalar relativistic approaches within the framework of density functional theory. [Pb(H(2)O)](2+) is found C(2v) at any level of calculations whereas [Pb(OH)](+) can be found bent or linear depending of the computational methodology used. When C(s) is found the barrier to inversion through the C(infinityv) structure is very low, and can be overcome at high enough temperature, making the molecule floppy. In order to get a better understanding of the bonding occurring between the Pb(2+) cation and the H(2)O and OH(-) ligands, natural bond orbital and atoms-in-molecule calculations have been performed. These approaches are supplemented by a topological analysis of the electron localization function. Finally, the description of these complexes is refined using constrained-space orbital variation complexation energy decompositions. PMID- 16689576 TI - Multireference configuration interaction study of the electronic states of ZrC. AB - The potential energy curves and spectroscopic constants of the ground and 32 low lying electronic states of ZrC have been studied by employing multireference configuration interaction methods, in conjunction with relativistic effective core potentials and 5s3p3d1f, 3s3p1d basis sets con Zr and C, respectively. We have determined that the ground state is (3)Sigma(+). However there are two low lying (1)Sigma(+) states (below 5000 cm(-1)) which strongly interact resulting in avoided crossings. The lowest (1)Sigma(+) state corresponds to a combination of 1sigma(2) Xsigma(2) 1pi(4) configurations whereas the second is an open shell singlet 1sigma(2) 2sigma(1) 3sigma(1) 1pi(4). Several avoided crossings were observed, for (1)Pi, (3)Pi, (1)Delta, (3)Sigma(+), and (3)Delta states. We have identified (3)Pi and (1)Pi lying at 4367 and 5797 cm(-1), respectively. The results are in good agreement with the recent experimental findings of Rixon et al. [J. Mol. Spectrosc. 228, 554 (2004)], and indicate that the (3)Pi (3)Sigma(+), and (1)Pi-(1)Sigma(+), bands located between 16 000-19 000 cm(-1) are extremely complex due to near degeneracy of several (1)Pi and (3)Pi states. We also have identified a (1)Sigma(+) state in the same region that may interfere with the (1)Pi emission bands. The present results not only shed further light into the spectra of ZrC but also predict yet to be observed systems. PMID- 16689577 TI - Collision-induced alignment of H2O+ drifting in helium. AB - The collision-induced alignment of H(2)O(+) drifting in helium is studied with a molecular dynamics method that has been extended to treat nonlinear rigid ions. Rotational distribution functions and averaged quantities are presented in terms of the rho formalism [M. Thachuk, Phys. Rev. A 72, 032722 (2005)], and it is shown that this description gives a very good agreement with simulation results. In addition to velocity and angular velocity autocorrelation functions, a velocity-angular velocity cross correlation function is introduced. This cross correlation function provides insight into the dynamical nature of the alignment mechanism. PMID- 16689578 TI - Water solvent effect on the first hyperpolarizability of p-nitrophenol and p nitrophenylphosphate: a time-dependent density functional study. AB - The first hyperpolarizabilities of p-nitrophenol and p-nitrophenylphosphate have been investigated in vacuum and in neutral aqueous solution by means of time dependent density functional theory. The calculated excited states and hyperpolarizabilities obtained for these systems and for the molecules of phenol, nitrobenzene, and p-nitroaniline in vacuum match well with the experimental trends. The water solvent has been described by the conductorlike screening model and has been completed by water molecules interacting by hydrogen bonds with the solute. The results show a significant effect of the solvent on the first hyperpolarizability. In particular, the hyperpolarizability of p nitrophenylphosphate (6.78 x 10(-30) esu) in vacuum is only 1.2 times larger than p-nitrophenol (5.63 x 10(-30) esu), whereas it is almost twice higher in aqueous environment, 12.6 x 10(-30) and 6.5 x 10(-30) esu, respectively. This difference in the nonlinear response in neutral water makes the p-nitrophenylphosphate substrate a suitable probe for measuring the activity of alkaline phosphatase enzymes. PMID- 16689579 TI - Implicit versus explicit solvent in free energy calculations of enzyme catalysis: Methyl transfer catalyzed by catechol O-methyltransferase. AB - We compare free energy calculations for the methyl transfer reaction catalyzed by catechol O-methyltransferase using the quantum mechanical/molecular mechanical free energy method with implicit and explicit solvents. An analogous methylation reaction in a solution is also studied. For the explicit solvent model, we use the three-point transferable intermolecular potential model, and for the implicit model, we use the generalized Born molecular volume model as implemented in CHARMM. We find that activation and reaction free energies calculated with the two models are very similar, despite some structural differences that exist. A significant change in the polarization of the environment occurs as the reaction proceeds. This is more pronounced for the reaction in a solution than for the enzymatic reaction. For the enzymatic reaction, most of the changes take place in the protein rather than in the solvent, and, hence, the benefit of having an instantaneous relaxation of the solvent degrees of freedom is less pronounced for the enzymatic reaction than for the reaction in a solution. This is a likely reason why energies of the enzyme reaction are less sensitive to the choice of atomic radii than are energies of the reaction in a solution. PMID- 16689580 TI - The bend angle of water in ice Ih and liquid water: The significance of implementing the nonlinear monomer dipole moment surface in classical interaction potentials. AB - The implementation of the physically accurate nonlinear dipole moment surface of the water monomer in the context of the Thole-type, polarizable, flexible interaction potential results in the only classical potential, which, starting from the gas phase value for the bend angle (104.52 degrees), reproduces its experimentally observed increase in the ice Ih lattice and in liquid water. This is in contrast to all other classical potentials to date, which predict a decrease of the monomer bend angle in ice Ih and in liquid water with respect to the gas phase monomer value. Simulations under periodic boundary conditions of several supercells consisting of up to 288 molecules of water used to sample the proton disorder in the ice Ih lattice yield an average value of vartheta(HOH)(I(h))=108.4 degrees +/-0.2 degrees for the minimized structures (T=0 K) and 108.1 degrees +/-2.8 degrees at T=100 K. Analogous simulations for liquid water predict an average value of vartheta(HOH)(liquid)=106.3 degrees +/ 4.9 degrees at T=300 K. The increase of the monomer bend angle of water in condensed environments is attributed to the use of geometry-dependent charges that are used to describe the nonlinear character of the monomer's dipole moment surface. Our results suggest a new paradigm in the development of classical interaction potential models of water that can be used to describe condensed aqueous environments. PMID- 16689581 TI - Study of multipole contributions to the structure of water around ions in solution using the soft sticky dipole-quadrupole-octupole (SSDQO) model of water. AB - The solvation of ions in the soft sticky dipole-quadrupole-octupole (SSDQO) model for liquid water is presented here. This new potential energy function for liquid water describes water-water interactions by a Lennard-Jones term plus a sticky potential consisting of an approximate moment expansion with point dipole, quadrupole, and octupole moments. The SSDQO potential energy function using the moments from extended simple point charge (SPC/E), TIP3P, or TIP5P reproduces the pair potential energy functions and radial distribution functions of the respective multipoint model but it is much faster than even the three-point models. Here, the solvation of ions in SSDQO water is studied using ion-water potential energy functions consisting of moment expansions up to the charge quadrupole term, up to the charge-octupole term, and up to an approximate charge hexadecapole term using the moments of SPC/E water. The radial distributions from Monte Carlo simulations show the best agreement with the results for ions in SPC/E water for the expansion up to the charge-hexadecapole term. Thus, the best results are obtained when the water-water and ion-water potentials are exact up to the 1r(4) term and also contain an approximate 1r(5) term. Overall, the simplicity, efficiency, and accuracy of the SSDQO potential make it potentially very useful for computer simulations of aqueous solvation. PMID- 16689582 TI - Cooperativity in ionic liquids. AB - Cooperativity in ionic liquids is investigated by means of static quantum chemical calculations. Larger clusters of the dimethylimidazolium cation paired with a chloride anion are calculated within density functional theory combined with gradient corrected functionals. Tests of the monomer unit show that density functional theory performs reasonably well. Linear chain and ring aggregates have been considered and geometries are found to be comparable with liquid phase structures. Cooperative effects occur when the total energy of the oligomer differs from a simple sum of monomer energies. Cooperative effects have been found in the structural motifs examined. A systematic study of linear chains of increasing length (up to nine monomer units) has shown that cooperativity plays a more important role than expected and is stronger than in water. The Cl...H distance of the chloride to the most acidic proton increases with an increasing number of monomer units. The average bond distance approaches 218.9 pm asymptotically. The dipole moment grows almost linearly and the dipole moment per monomer unit reaches the asymptotic value of 16.3 D. The charge on the chloride atoms decreases with an increasing chain length. In order to detect local hydrogen bonding in the clusters a new parametrization of the shared-electron number method is introduced. We find decreasing hydrogen bond energies with an increasing cluster size for both the first hydrogen bond to the most acidic proton and the average hydrogen bond. PMID- 16689583 TI - Monte Carlo simulation of crystal-fluid coexistence states in the hard-sphere system under gravity with stepwise control. AB - Monte Carlo (MC) simulations were performed for hard spheres (with diameter sigma and mass m) placed between well-separated upper and lower hard walls. A periodic boundary condition was imposed in the horizontal direction. The system was exposed to the gravitational field with the acceleration due to gravity g. After preparing a melt as the initial state, g was increased stepwise up to mgsigma/k(B)T(identical with g(*))=1.5 or 2.0 with an increment Deltag(*) = 0.1; k(B)T is the temperature multiplied by Boltzmann's constant. We maintained g(*) at each value for 2.0 x 10(5) MC cycles. The transition of the system into a metastable state such as a polycrystalline state due to trapping phenomena was successfully avoided. A monotonic increase and subsequent saturation were observed for the development of the crystalline region formed at the bottom of the system. The development of this region accompanied a shrinkage of the defective (or less ordered) crystalline region that was formed between the bottom region and the fluid phase. As the development of the bottom region almost saturated, the defective region grew upward again. PMID- 16689584 TI - Ab initio study of structural and electronic properties of SrTiO3 (001) oxygen vacancy surfaces. AB - First-principles calculations are employed to study the surface relaxation and electronic structure of the fully relaxed SrTiO(3) (001) oxygen-vacancy surfaces with both Sr and Ti terminations. In contrast to the perfect surface, the larger surface rumples and smaller interlayer distances have been found. Some in-gap Ti 3d states at about -1.13 eV below the Fermi level were observed in the Ti terminated surface caused by oxygen vacancies. For the Sr-terminated oxygen vacancy surface, some in-gap Ti 3d states move into the bulk midgap region to become partially occupied. These theoretical results are in agreement with the experimental data. PMID- 16689585 TI - Ab initio studies of layering behavior of liquid sodium surfaces and interfaces. AB - We have studied the liquid surface of sodium with extensive ab initio molecular dynamics simulations based on ensemble density-functional theory. We find clear evidence of layering in the direction perpendicular to the surface that persists to temperatures more than 100 K above the melting point. We also observe clear Friedel oscillations in the electronic density response to the presence of a surface, but their direct effect on atomic layering is ruled out. A careful finite-size effect analysis accompanies our results, showing that liquid slabs 20 25 A thick capture the essential details of the surface structure. We conclude that geometrical confinement is the common cause for layer formation, which is similar to what happens at a liquid-solid interface: at a free liquid surface, the rapid decay of the electronic density from the bulk liquid value to zero in the vapor forms a hard wall against which the atoms pack. Finally, we predict x ray reflectivities from ab initio molecular dynamics data that include some of the large surface-normal wave vector-transfer regions that, for alkali metals, are not accessible to experiments. PMID- 16689586 TI - Friction and tribochemical reactions occurring at shearing interfaces of nanothin silver films on various substrates. AB - The tribological and tribochemical properties of 5-10 nm thick Ag films sliding on various metal and inorganic substrates were measured using a surface forces apparatus coupled with ex situ x-ray photoelectron spectroscopy. We observed enhanced chemical reactivity in the sheared regions compared to the unsheared regions, which we attribute to significant frictional heating in agreement with two recent simulations. It is also found that the initial topography (roughness) of the films plays a significant role in determining the friction, wear, and tribochemical reactions. The larger the surface roughness, the larger are the friction coefficients. Initially smooth surfaces, forming large continuous junctions, create large wear debris particles; whereas initially rough surfaces, forming many small junctions, create many small particles. Even though initially smooth surfaces are chemically less reactive than initially rough surfaces, they are tribochemically more reactive, forming two to three times more oxides of silver during shearing than under static conditions and organometals of silver that under normal (static) conditions do not form. The chemical reactions observed cannot be explained without including the tribological processes, such as the local stresses, temperature rises, and type of wear produced by the shearing surfaces. PMID- 16689587 TI - Control of morphology in efficient photovoltaic diodes from discotic liquid crystals. AB - We report on the role of morphology in photovoltaic diodes with blend active layers composed of perylene tetracarboxdiimide (EPPTC) and hexabenzocoronene (HBC) derivatives as electron and hole acceptors. Controlled annealing of HBC:EPPTC films while in conformal contact with a flat elastomeric stamp improves photovoltaic response, leading to an external quantum efficiency of 29.5% at 460 nm and an open-circuit voltage of 0.70 V. The improved performance is attributed in part to larger crystalline domains following annealing. The elastomeric stamp restricts the top surface of the thin film during annealing, leading to low surface roughness, while also allowing for greater vertical stratification of the components in the bulk. Blended HBC:EPPTC films also exhibit an unique optical absorption feature near 590 nm, which we attribute to an altered crystalline packing of EPPTC in the presence of HBC. The significance of the local structure at molecular heterojunctions in blended active layer photovoltaic diodes is discussed. PMID- 16689588 TI - Structural, electronic, and magnetic properties of manganese-doped Zn12O12 clusters: a first-principles study. AB - A first-principles study has been performed to evaluate the structural, electronic, and magnetic properties of Zn(12)O(12) clusters doped with one or two Mn atoms. The substitutional, exohedral, and endohedral dopings are taken into account. For the monodoped clusters, the substitutional isomer is most energetically favorable, and an exohedral isomer may appear as a low-lying metastable state. All isomers present 5 mu(B) magnetic moment that is mainly contributed by the Mn-3d component. For the bidoped clusters, the antiferromagnetic state is degenerate with the ferromagnetic state at larger Mn Mn distance (>5 A), while it is more energetically favorable at smaller Mn-Mn distance. Thus, the cohesion of bidoped isomer is sensitive to the magnetic coupling or chemical bonding. The endohedral bidoped isomer is found to be a stable local minimum, and the direct Mn-Mn interaction causes the reduction of local magnetic moment of Mn to about 4 mu(B). PMID- 16689589 TI - A nonmonotonic dependence of standard rate constant on reorganization energy for heterogeneous electron transfer processes on electrode surface. AB - In the present work a nonmonotonic dependence of standard rate constant (k(0)) on reorganization energy (lambda) was discovered qualitatively from electron transfer (Marcus-Hush-Levich) theory for heterogeneous electron transfer processes on electrode surface. It was found that the nonmonotonic dependence of k(0) on lambda is another result, besides the disappearance of the famous Marcus inverted region, coming from the continuum of electronic states in electrode: with the increase of lambda, the states for both Process I and Process II ET processes all vary from nonadiabatic to adiabatic state continuously, and the lambda dependence of k(0) for Process I is monotonic thoroughly, while for Process II on electrode surface the lambda dependence of k(0) could show a nonmonotonicity. PMID- 16689590 TI - Ultrafast electron diffraction: excited state structures and chemistries of aromatic carbonyls. AB - The photophysics and photochemistry of molecules with complex electronic structures, such as aromatic carbonyls, involve dark structures of radiationless processes. With ultrafast electron diffraction (UED) of isolated molecular beams it is possible to determine these transient structures, and in this contribution we examine the nature of structural dynamics in two systems, benzaldehyde and acetophenone. Both molecules are seen to undergo a bifurcation upon excitation (S(2)). Following femtosecond conversion to S(1), the bifurcation leads to the formation of molecular dissociation products, benzene and carbon monoxide for benzaldehyde, and benzoyl and methyl radicals for acetophenone, as well as intersystem crossing to the triplet state in both cases. The structure of the triplet state was determined to be "quinoidlike" of pipi(*) character with the excitation being localized in the phenyl ring. For the chemical channels, the product structures were also determined. The difference in photochemistry between the two species is discussed with respect to the change in large amplitude motion caused by the added methyl group in acetophenone. This discussion is also expanded to compare these results with the prototypical aliphatic carbonyl compounds, acetaldehyde and acetone. From these studies of structural dynamics, experimental and theoretical, we provide a landscape picture for, and the structures involved in, the radiationless pathways which determine the fate of molecules following excitation. For completeness, the UED methodology and the theoretical framework for structure determination are described in this full account of an earlier communication [J. S. Feenstra et al., J. Chem. Phys. 123, 221104 (2005)]. PMID- 16689591 TI - Coarse-grained interaction potentials for anisotropic molecules. AB - We have proposed an efficient parametrization method for a recent variant of the Gay Berne potential for dissimilar and biaxial particles [Phys. Rev. E 67, 041710 (2003)] and demonstrated it for a set of small organic molecules. Compared with the previously proposed coarse-grained models, the new potential exhibits a superior performance in close contact and large distant interactions. The repercussions of thermal vibrations and elasticity have been studied through a statistical method. The study justifies that the potential of mean force is representable with the same functional form, extending the application of this coarse-grained description to a broader range of molecules. Moreover, the advantage of employing coarse-grained models over truncated atomistic summations with large distance cutoffs has been briefly studied. PMID- 16689592 TI - Vibrational and electron paramagnetic resonance properties of free and MgO supported AuCO complexes. AB - The bonding, spin density related properties, and vibrational frequency of CO bound to single Au atom in the gas-phase or supported on MgO surfaces have been investigated with a variety of computational methods and models: periodic plane waves calculations have been compared with molecular approaches based on atomic orbital basis sets; pseudopotential methods with all electron fully relativistic calculations; various density functional theory (DFT) exchange-correlation functionals with the unrestricted coupled-cluster singles and doubles with perturbative connected triples [CCSD(T)]. AuCO is a bent molecule but the potential for bending is very soft, and small changes in the bond angle result in large changes in the CO gas-phase vibrational frequency. At the equilibrium geometry the DFT calculated vibrational shift of CO with respect to the free molecule is about -150 cm(-1), whereas smaller values -60-70 cm(-1) are predicted by the more accurate CCSD(T) method. These relatively large differences are due to the weak and nonclassic bonding in this complex. Upon adsorption on MgO, the CO vibrational shift becomes much larger, about -290 cm(-1), due to charge transfer from the basic surface oxide anion to AuCO. This large redshift is predicted by all methods, and is fully consistent with that measured for MgOAuCO complexes. The strong influence of the support on the AuCO bonding is equally well described by all different approaches. PMID- 16689593 TI - Phosphonate self-assembled monolayers on aluminum surfaces. AB - Substrates of aluminum (Al) deposited by physical vapor deposition onto Si substrates and then chemically reacted with perfluorodecylphosphonic acid (PFDPAlSi), decylphosphonic acid (DPAlSi), and octadecylphosphonic acid (ODPAlSi) were studied by x-ray photoelectron spectroscopy (XPS), contact angle measurements, atomic force microscopy (AFM), and friction force microscopy, a derivative of AFM, to characterize their surface chemical composition, roughness, and micro-/nanotribological properties. XPS analysis confirmed the presence of perfluorinated and nonperfluorinated alkylphosphonate molecules on the PFDPAlSi, DPAlSi, and ODPAlSi. The sessile drop static contact angle of pure water on PFDPAlSi was typically more than 130 degrees and on DPAlSi and ODPAlSi typically more than 125 degrees indicating that all phosphonic acid reacted AlSi samples were very hydrophobic. The surface roughness for PFDPAlSi, DPAlSi, ODPAlSi, and bare AlSi was approximately 35 nm as determined by AFM. The surface energy for PFDPAlSi was determined to be approximately 11 mNm by the Zisman plot method compared to 21 and 20 mNm for DPAlSi and ODPAlSi, respectively. Tribology involves the measure of lateral forces due to friction and adhesion between two surfaces. Friction, adhesion, and wear play important roles in the performance of micro-/nanoelectromechanical systems. PFDPAlSi gave the lowest adhesion and coefficient of friction values while bare AlSi gave the highest. The adhesion and coefficient of friction values for DPAlSi and ODPAlSi were comparable. PMID- 16689594 TI - Study of electronic and spectroscopic properties on a newly synthesized red fluorescent material. AB - The ground state (S(0)) and the lowest singlet excited state (S(1)) of a newly synthesized red fluorescent material, 2-[3-(2-{4-[(2-Hydroxy-ethyl)-methyl-amino] phenyl}-vinyl)-5,5-dimethyl-cyclohex-2-enylidene]-malononitrile (A31), are investigated. The S(0) and S(1) geometries are optimized at the ab initio Hartree Fock and the singles configuration interaction (CIS) levels of theory, respectively. The CIS and semiempirical Zerner's Intermediate Neglect of Differential Overlap (ZINDO) methods provide the results for the absorption (S(0) ->S(1)) and emission (S(1)-->S(0)) transition energies. The Stokes shifts calculated at the CIS and ZINDO levels of theory are obtained. The absorption spectra in various solvents are calculated using the time-dependent density functional theory method in combination with the polarized continuum model, which are in very good agreement with our experimental measurements. The solvent effects are discussed. PMID- 16689595 TI - Effects of adsorbed water layer structure on adhesion force of silicon oxide nanoasperity contact in humid ambient. AB - The origin of the large relative-humidity (RH) dependence of the adhesion force in the single-asperity contact between silicon oxide surfaces is elucidated. As RH increases, the adhesion force measured with an atomic force microscopy (AFM) initially increases, reaches a maximum, and then decreases at high RH. The capillary force alone cannot explain the observed magnitude of the RH dependence. The origin of the large RH dependence is due to the presence of an icelike structured water adsorbed at the silicon oxide surface at room temperature. A solid-adsorbate-solid model is developed calculating the contributions from capillary forces, van der Waals interactions, and the rupture of an ice-ice bridge at the center of the contact region. This model illustrates how the structure, thickness, and viscoelastic behavior of the adsorbed water layer influence the adhesion force of the silicon oxide nanoasperity contact. PMID- 16689596 TI - Ab initio based tight-binding molecular dynamics simulation of the sticking and scattering of O2Pt(111). AB - The sticking and scattering of O(2)Pt(111) has been studied by tight-binding molecular dynamics simulations based on an ab initio potential energy surface. We focus, in particular, on the sticking probability as a function of the angle of incidence and the energy and angular distributions in scattering. Our simulations provide an explanation for the seemingly paradox experimental findings that adsorption experiments suggest that the O(2)Pt(111) interaction potential should be strongly corrugated while scattering experiments indicate a rather small corrugation. The potential energy surface is indeed strongly corrugated which leads to a pronounced dependence of the sticking probability on the angle of incidence. The scattered O(2) molecules, however, experience a rather flat surface due to the fact that they are predominantly scattered at the repulsive tail of the potential. PMID- 16689597 TI - Hydrogen bond dynamics and microscopic structure of confined water inside carbon nanotubes. AB - We have investigated the density and temperature dependences of microscopic structure and hydrogen bond dynamics of water inside carbon nanotubes (CNTs) using molecular dynamics simulation. The CNTs are treated as rigid, and smoothly truncated extended simple point charge water model is adopted. The results show that as the overall density increases, the atomic density profiles of water inside CNTs become sharper, the peaks shift closer to the wall, and a new peak of hydrogen atomic density appears between the first (outermost) and second layer. The intermittent hydrogen bond correlation function C(HB)(t) of water inside CNTs decays slower than that of bulk water, and the rate of decay decreases as the tube diameter decreases. C(HB)(t) clearly decays more slowly for the first layer of water than for other regions inside CNTs. The C(HB)(t) of the interlayer hydrogen bonds decays faster than those of the other regions and even faster than that of the bulk water. On the other hand, the hydrogen bond lifetimes of the first layer are shorter than those of the inner layer(s). Interlayer hydrogen bond lifetimes are clearly shorter than those of the constituent layers. As a whole, the hydrogen bond lifetimes of water inside CNTs are shorter than those of bulk water, while the relaxation of C(HB)(t) is slower for the confined water than for bulk water. In other words, hydrogen bonds of water inside CNTs break more easily than those of bulk water, but the water molecules remain in each other's vicinity and can easily reform the bonds. PMID- 16689598 TI - Conformational sampling via a self-regulating effective energy surface. AB - The difficulty of efficiently sampling the phase space of complex systems with rough energy surfaces is well known. Typical solutions to the problem involve accelerating the crossing of barriers, but such methods often have the secondary problem that the low-energy states of interest are inadequately sampled, unless the parameters of the search algorithm are modified as the system evolves. A method is presented to improve the sampling with particular emphasis on the low energy conformations, which make the most important contributions to the thermodynamics of the system. The algorithm proposed here samples the details of the minima, while easily surmounting barriers. This is achieved by introducing a self-regulating sampling variable which depends on the current state of the system. Two replicas of the system are introduced and the sampling variable is treated as a particle coupled to the physical system. The method is illustrated with a simple model system and is applied to the realistic example of barrier crossing in a protein-ligand complex. PMID- 16689599 TI - Vitrification of polymer solutions as a function of solvent quality, analyzed via vapor pressures. AB - Vapor pressures (headspace sampling in combination with gas chromatography) and glass transition temperatures [differential scanning calorimetry (DSC)] have been measured for solutions of polystyrene (PS) in either toluene (TL) (10-70 degrees C) or cyclohexane (CH) (32-60 degrees C) from moderately concentrated solutions up to the pure polymer. As long as the mixtures are liquid, the vapor pressure of TL (good solvent) is considerably lower than that of CH (theta solvent) under other identical conditions. These differences vanish upon the vitrification of the solutions. For TL the isothermal liquid-solid transition induced by an increase of polymer concentration takes place within a finite composition interval at constant vapor pressure; with CH this phenomenon is either absent or too insignificant to be detected. For PS solutions in TL the DSC traces look as usual, whereas these curves may become bimodal for solutions in CH. The implications of the vitrification of the polymer solutions for the determination of Flory-Huggins interaction parameters from vapor pressure data are discussed. A comparison of the results for TL/PS with recently published data on the same system demonstrates that the experimental method employed for the determination of vapor pressures plays an important role at high polymer concentrations and low temperatures. PMID- 16689600 TI - Optimized parallel tempering simulations of proteins. AB - We apply a recently developed adaptive algorithm that systematically improves the efficiency of parallel tempering or replica exchange methods in the numerical simulation of small proteins. Feedback iterations allow us to identify an optimal set of temperatures/replicas which are found to concentrate at the bottlenecks of the simulations. A measure of convergence for the equilibration of the parallel tempering algorithm is discussed. We test our algorithm by simulating the 36 residue villin headpiece subdomain HP-36 where we find a lowest-energy configuration with a root-mean-square deviation of less than 4 A to the experimentally determined structure. PMID- 16689601 TI - Recognition of complex patterned substrates by heteropolymer chains consisting of multiple monomer types. AB - We propose a statistical mechanical model of surface pattern recognition by heteropolymers with quenched monomer sequence distribution. The chemically heterogeneous pattern consists of different adsorption sites specifically distributed on a surface. The heteropolymer sequence is complementary with respect to the pattern. The concepts of recognition probability and recognition temperature are introduced. The algorithm for calculating the recognition probability is based on efficient recurrence procedures for evaluating the single chain partition function of a chain macromolecule consisting of multiple monomer types, which interact with multiple types of adsorption sites. The temperature dependencies of the recognition probability are discussed. We address the critical role of the commensurability between the heteropolymer sequence and the distribution of the surface adsorbing sites on the polymer adsorption. Also, we address the question of how many types of monomer units in the heteropolymer are required for unambiguous recognition of compact target patterns. It is shown that perfect pattern recognition can be achieved for the strong-adsorption regime in the case of specifically structured compact patterns with multifunctional adsorption sites and heteropolymers with multiple monomer types when the degeneracy of the ground state is suppressed. The pattern recognition ability increases with the number of different types of monomer units and complementary adsorption sites. For random heteropolymers and patterns, the free energy change associated with the recognition process decreases linearly with increasing this number. Correlated random heteropolymers are capable of recognizing related patterns on a random background. PMID- 16689602 TI - Spacers' role in the dynamics of hyperbranched polymers. AB - We investigate hyperbranched polymers (HBPs) and highlight the relation between their architecture and their viscoelastic behavior, while paying special attention to the role of the chainlike spacer segments between branching points. For this we study the dynamics of HBP in solution, based on the generalized Gaussian structure formalism, an extension of the Rouse model, which disregards hydrodynamical and excluded volume effects. For HBP the dynamical effects display, beside the obvious contributions of localized modes on the spacers, also remarkable features, as we highlight based on the exact renormalization procedure recently developed by us in J. Chem. Phys. 123, 034907 (2005). We exemplify these features by analyzing the dynamics of randomly linked star polymers and study the impact both of the length and of the spacers' mobility on the normal modes' spectra. We compute these modes both by numerical diagonalization and also by employing our renormalization procedure; the excellent agreement between these methods allows us to extend the range of investigations to very large HBP. PMID- 16689603 TI - Excluded volume effect on confined polymer translocation through a short nanochannel. AB - We simulated the translocation process of a polymer chain from a source container to a drain container through a short nanochannel. We utilized the bond fluctuation model coupled with Monte Carlo dynamics in our simulations. The calculation results show that the excluded volume effect significantly affects the polymer's translocation time tau. This time depends nonmonotonically on the polymer length N. For a fixed nanochannel length, tau decreases when the polymer length increases. tau, however, increases when the polymer length exceeds a certain threshold. This observation differs from those predicated for a Gaussian chain. In this paper, we will further present our findings to explain this phenomenon. The knowledge we gain from this research can enhance the understanding of complex transport processes in many biological systems. PMID- 16689604 TI - Local and chain dynamics in miscible polymer blends: A Monte Carlo simulation study. AB - Local chain structure and local environment play an important role in the dynamics of polymer chains in miscible blends. In general, the friction coefficients that describe the segmental dynamics of the two components in a blend differ from each other and from those of the pure melts. In this work, we investigate polymer blend dynamics with Monte Carlo simulations of a generalized bond fluctuation model, where differences in the interaction energies between nonbonded nearest neighbors distinguish the two components of a blend. Simulations employing only local moves and respecting a no bond crossing condition were carried out for blends with a range of compositions, densities, and chain lengths. The blends investigated here have long time dynamics in the crossover region between Rouse and entangled behavior. In order to investigate the scaling of the self-diffusion coefficients, characteristic chain lengths N(c) are calculated from the packing length of the chains. These are combined with a local mobility mu determined from the acceptance rate and the effective bond length to yield characteristic self-diffusion coefficients D(c)=muN(c). We find that the data for both melts and blends collapse onto a common line in a graph of reduced diffusion coefficients DD(c) as a function of reduced chain length NN(c). The composition dependence of dynamic properties is investigated in detail for melts and blends with chains of length N=20 at three different densities. For these blends, we calculate friction coefficients from the local mobilities and consider their composition and pressure dependence. The friction coefficients determined in this way show many of the characteristics observed in experiments on miscible blends. PMID- 16689605 TI - A microscopic look at the reinforcement of silica-filled rubbers. AB - The deformed structure of silica-filled elastomers under uniaxial strain has been investigated using a combination of both small angle x-ray scattering and small angle neutron scattering methods. Using an extraction procedure and taking into account the two-phase nature of these polymer-based composites, the single chain scattering behavior as well as filler properties could be obtained uniquely on identical samples. For the first time the deformation of the rubbery matrix on the length scale of the network chain in a filled rubber could be determined and therewith the importance of matrix overstrain for the mechanical properties was estimated. Additionally, the determination of filler deformation and filler destruction presents microscopic details of the mechanisms of filler networking and the stress-softening Mullins effect. PMID- 16689606 TI - On the analysis of Liquid Crystal NMR dipolar coupling data via mixed a priori maximum entropy methods. AB - In this Letter, a general expression is derived for the conformational distribution function of a molecule dissolved in an anisotropic condensed fluid medium by combining an a priori model with the maximum entropy principle applied to treatment of liquid crystalline-NMR data. The recently proposed additive potential maximum entropy (APME) method is recovered as a special case, when the AP is chosen as the a priori model and the orientational order is low. PMID- 16689610 TI - Inclusion of people with mental retardation and other developmental disabilities in communities of faith. AB - Our focus in this paper is on efforts to include persons with developmental disabilities in faith communities. We provide a review of the relevant literature on religious participation and faith communities for persons with disabilities and blend the limited data available on these topics with the perspectives of individuals whose efforts focus on these concerns. Topics explored are the implications of being part of the faith community in terms of its impact on quality of life, the barriers to inclusion in such communities, strategies for overcoming these barriers, and special considerations for adults with mental retardation or other developmental disabilities. Discussion of the implications for enhancing inclusion in faith communities is provided. PMID- 16689611 TI - Assessment instruments measuring malingering used with individuals who have mental retardation: potential problems and issues. AB - Malingering, the exaggeration or fabrication of physical and/or psychological symptoms, can threaten the psychological assessment process (American Psychiatric Association, 2000). To enhance the validity of psychological evaluations, researchers have studied trends in malingering and developed instruments for its detection (Rogers, Bagby, & Dickens, 1992; Tombaugh, 1996). These instruments, however, may not be appropriate for individuals with significant subaverage intellectual functioning. Four instruments assessing malingering, frequently used in forensic evaluations, were administered to individuals with mental retardation. Results show that by utilizing established cut-off scores, we were able to classify a significant percentage of participants as "malingering," in spite of directions to perform optimally. Practical implications as well as directions for future research are discussed. PMID- 16689612 TI - Comparison of reports by relatives and staff on living conditions of adults with intellectual disabilities. AB - Proxies typically serve as information providers in studies of persons with intellectual disabilities. However, little is known about the concordance between different proxy categories and how proxy characteristics influence the information provided. We compared 89 pairs of relative and staff reports on the living conditions of persons with intellectual disabilities, using percentage agreement and Cohen's kappa statistics. Results demonstrate differences between relative and staff reports for most of the domains investigated, with moderate agreement for objective items and fair agreement for subjective items. Relative and staff proxies contributed different information related to diverse viewpoints and varying types of information. Thus, we suggest that information provided by proxies should not be treated as being interchangeable but, rather, as complementary. PMID- 16689613 TI - Health insurance coverage of direct support workers in the developmental disabilities field. AB - There is mounting evidence that employer-provided health insurance is an important factor in recruiting and retaining a competent and motivated direct support workforce within health and human services occupations. A review of the literature in this area, including new information related to the developmental disabilities field, is presented to assist nonprofit employers and government officials in designing initiatives to address increasing health care costs. Approaches to financing health coverage for frontline staff and a new program in New York that will provide subsidies to agencies to enhance existing coverage are discussed. PMID- 16689614 TI - "Who would I be without Danny?" phenomenological case study of an adult sibling. AB - How is life different for someone who grows up with a sibling who has mental retardation? In this phenomenological case study, I profile the life experiences of a 39-year-old woman as sibling to a 35-year-old man with Down syndrome, using her own words as she explored her sibling experience and how this relationship shaped her identity, experiences, and life choices. Emerging themes, including her self-perception and perception of family members, her identity formation, and the impact of the experience are discussed. A number of possible variables for future quantitative research are suggested based on her experiences, including increased empathy in typically developing siblings and the sibling with disability as a barometer of other people's traits and their impact on the formation of other significant relationships. PMID- 16689615 TI - Christmas in purgatory: a retrospective look. PMID- 16689616 TI - Leaving the garden: reconsidering Henry Herbert Goddard's exodus from the Vineland Training School. PMID- 16689619 TI - The Supreme Court, "Our Town," and disability policy: boardrooms and bedrooms, courtrooms and cloakrooms. AB - In this discussion of recent key disability-related decisions of the U.S. Supreme Court (1995- 2004), we (a) assess whether the Court has supported or undermined certain core concepts of disability policy and (b) examine how the Court balances the comparative rights of those with and those without disabilities. In cases involving employment discrimination, family law, and access to courts and other public decision-making entities, the Court adopts an idealized version of a previous America. We explain the Court's "reverie" for that version, resorting to the constructs known as (a) compelled confrontation and remission to majoritarian processes and (b) advancement of personal relationships. PMID- 16689620 TI - Development of a submaximal test to predict elliptical cross-trainer VO2max. AB - The purpose of this study was to develop an equation to predict VO2max from a submaximal elliptical cross-trainer test. Fifty-four apparently healthy subjects (25 men and 29 women, mean +/- SD age: 29.5 +/- 7.1 years, height: 173.3 +/- 12.6 cm, weight: 72.3 +/- 7.9 kg, percent body fat: 17.3 +/- 5.0%, and elliptical cross-trainer VO2max: 43.9 +/- 7.2 ml x kg(-1) x min(-1)) participated in the study and were randomly assigned to an original sample group (n = 40) and a cross validation group (n = 14). Each subject completed an elliptical cross-trainer submaximal (3 5-minute submaximal stages) and a VO2max test on the same day, with a 15-minute rest period in between. Stepwise multiple regression analyses were used to develop an equation for estimating elliptical cross-trainer VO2max from the data of the original sample group. The accuracy of the equation was tested by using data from the cross-validation group. Because there was no shrinkage in R2 between the original sample group and the cross-validation group, data were combined in the final prediction equation (R2 = 0.732, standard error of the estimate = 3.91 ml x kg(-1) x min(-1), p < 0.05): VO2max = 73.676 + 7.383(gender) - 0.317(weight) + 0.003957(age x cadence) - 0.006452(age x heart rate at stage 2). The correlation coefficient between the predicted and measured VO2max values was r = 0.86. Dependent t-tests resulted in no significant differences (p > 0.05) between predicted (43.8 ml x kg(-1) x min(-1)) and measured (43.9 ml x kg(-1) x min(-1)) VO2max measurements. Results indicate that the protocol and equation developed in the current study can be used by exercise professionals to provide acceptably accurate estimates of VO2max in non-laboratory-based settings. PMID- 16689621 TI - Aerobic fitness and yo-yo continuous and intermittent tests performances in soccer players: a correlation study. AB - Yo-yo tests are very popular in soccer; however, no study has addressed details of their relation to canonical aspects of aerobic fitness. Furthermore, no information is available on the effect of the individual levels of lower limbs' explosive strength on yo-yo tests in soccer players. The purpose of this study was to examine the physiological determinants of Yo-yo Endurance Test Level 2 (YYETL2) and Yo-yo Intermittent Recovery Test Level 1 (YYIRTL1) in soccer players. Twenty-four soccer players (body mass, 74.6 +/- 8.5 kg; height, 178.1 +/ 4.5 cm; age, 25.6 +/- 5.1 years) were tested for VO2max and ventilatory threshold (VT) on a motorized treadmill. Lower-limb explosive strength was assessed using vertical countermovement jumps (CMJ) performed on a force platform. Results showed that YYETL2 and YYIRTL1 performances (m) were significantly related (r = 0.75, p = 0.00002). YYETL2 results were significantly related to VO2max, VTVO2, and speed at VT (r = 0.75, 0.76, and 0.83, respectively; p < 0.00002). Peak treadmill speed results were significantly related to YYETL2 and YYIRTL1 (r = 0.87 and 0.71, respectively; p < 0.0003). YYIRTL1 was related to CMJ peak power (r = 0.57; p = 0.003). These findings show that YYETL2 and YYIRTL1, although adopting similar starting and progression speeds, are influenced by different physiological variables. From these results, YYETL2 can be considered an aerobic fitness-related field test, whereas YYIRTL1 can be regarded as an aerobic-anaerobic, soccer-specific field test. PMID- 16689622 TI - Cardiorespiratory responses to Yo-yo Intermittent Endurance Test in nonelite youth soccer players. AB - This study examined the validity of the Yo-yo Intermittent Endurance Test (Level 1; YYIET) as indicator of aerobic power in youth soccer players. Cardiorespiratory responses were determined in 18 moderately trained nonelite youth soccer players (age, 16.6 +/- 0.8 years; height, 178.7 +/- 6.2 cm; body mass, 69.8 +/- 6.0 kg; VO2peak, 52.8 +/- 7.4 ml x kg(-1) x min(-1)) while performing the YYIET and an incremental treadmill test. Maximal heart rate (HRmax), respiratory exchange ratio (RER), O2 pulse, VO2peak, and maximal ventilation (VEmax) were measured. Group YYIET VO2peak, HRmax, RER, and O2 pulse were not significantly different from treadmill responses (p > 0.05). VEmax was significantly lower (p < 0.05) during the YYIET compared to the treadmill condition. No significant correlation was found between treadmill VO2peak and YYIET performance (p > 0.05). This study showed that the YYIET elicits peak VO2 and HR responses. However, YYIET performance results were not related to VO2peak measured in laboratory. Furthermore, the individual VO2peak reached during the TM did not reflect the VO2peak obtained during the YYIET, as shown by the large limits of agreement. As a consequence, compared to other shuttle run field tests, YYIET seems to be a weak indicator of aerobic power in youth moderately trained youth soccer player. PMID- 16689626 TI - Spinach on the ceiling: a theoretical chemist's return to biology. AB - I was born in Vienna and came to the United States as a refugee in October 1938. This experience played an important role in my view of the world and my approach to science: It contributed to my realization that it was safe to stop working in fields that I felt I understood and to focus on different areas of research by asking questions that would teach me and others something new. I describe my experiences that led me from chemistry and physics back to my first love, biology, and outline some of the contributions I have made as part of my ongoing learning experience. PMID- 16689623 TI - Neuronal oscillations enhance stimulus discrimination by ensuring action potential precision. AB - Although oscillations in membrane potential are a prominent feature of sensory, motor, and cognitive function, their precise role in signal processing remains elusive. Here we show, using a combination of in vivo, in vitro, and theoretical approaches, that both synaptically and intrinsically generated membrane potential oscillations dramatically improve action potential (AP) precision by removing the membrane potential variance associated with jitter-accumulating trains of APs. This increased AP precision occurred irrespective of cell type and--at oscillation frequencies ranging from 3 to 65 Hz--permitted accurate discernment of up to 1,000 different stimuli. At low oscillation frequencies, stimulus discrimination showed a clear phase dependence whereby inputs arriving during the trough and the early rising phase of an oscillation cycle were most robustly discriminated. Thus, by ensuring AP precision, membrane potential oscillations dramatically enhance the discriminatory capabilities of individual neurons and networks of cells and provide one attractive explanation for their abundance in neurophysiological systems. PMID- 16689627 TI - Computer-based design of novel protein structures. AB - Over the past 10 years there has been tremendous success in the area of computational protein design. Protein design software has been used to stabilize proteins, solubilize membrane proteins, design intermolecular interactions, and design new protein structures. A key motivation for these studies is that they test our understanding of protein energetics and structure. De novo design of novel structures is a particularly rigorous test because the protein backbone must be designed in addition to the amino acid side chains. A priori it is not guaranteed that the target backbone is even designable. To address this issue, researchers have developed a variety of methods for generating protein-like scaffolds and for optimizing the protein backbone in conjunction with the amino acid sequence. These protocols have been used to design proteins from scratch and to explore sequence space for naturally occurring protein folds. PMID- 16689629 TI - Evolutionary relationships and structural mechanisms of AAA+ proteins. AB - Complex cellular events commonly depend on the activity of molecular "machines" that efficiently couple enzymatic and regulatory functions within a multiprotein assembly. An essential and expanding subset of these assemblies comprises proteins of the ATPases associated with diverse cellular activities (AAA+) family. The defining feature of AAA+ proteins is a structurally conserved ATP binding module that oligomerizes into active arrays. ATP binding and hydrolysis events at the interface of neighboring subunits drive conformational changes within the AAA+ assembly that direct translocation or remodeling of target substrates. In this review, we describe the critical features of the AAA+ domain, summarize our current knowledge of how this versatile element is incorporated into larger assemblies, and discuss specific adaptations of the AAA+ fold that allow complex molecular manipulations to be carried out for a highly diverse set of macromolecular targets. PMID- 16689628 TI - Lessons from lactose permease. AB - An X-ray structure of the lactose permease of Escherichia coli (LacY) in an inward-facing conformation has been solved. LacY contains N- and C-terminal domains, each with six transmembrane helices, positioned pseudosymmetrically. Ligand is bound at the apex of a hydrophilic cavity in the approximate middle of the molecule. Residues involved in substrate binding and H+ translocation are aligned parallel to the membrane at the same level and may be exposed to a water filled cavity in both the inward- and outward-facing conformations, thereby allowing both sugar and H+ release directly into either cavity. These structural features may explain why LacY catalyzes galactoside/H+ symport in both directions utilizing the same residues. A working model for the mechanism is presented that involves alternating access of both the sugar- and H+-binding sites to either side of the membrane. PMID- 16689630 TI - Symmetry, form, and shape: guiding principles for robustness in macromolecular machines. AB - Computational studies of large macromolecular assemblages have come a long way during the past 10 years. With the explosion of computer power and parallel computing, timescales of molecular dynamics simulations have been extended far beyond the hundreds of picoseconds timescale. However, limitations remain for studies of large-scale conformational changes occurring on timescales beyond nanoseconds, especially for large macromolecules. In this review, we describe recent methods based on normal mode analysis that have enabled us to study dynamics on the microsecond timescale for large macromolecules using different levels of coarse graining, from atomically detailed models to those employing only low-resolution structural information. Emerging from such studies is a control principle for robustness in Nature's machines. We discuss this idea in the context of large-scale functional reorganization of the ribosome, virus particles, and the muscle protein myosin. PMID- 16689631 TI - Fusion pores and fusion machines in Ca2+-triggered exocytosis. AB - Exocytosis is initiated within a highly localized region of contact between two biological membranes. Small areas of these membranes draw close, molecules on the two surfaces interact, and structural transformations take place. Membrane fusion requires the action of proteins specialized for this task, and these proteins act as a fusion machine. At a critical point in this process, a fusion pore forms within the membrane contact site and then expands as the spherical vesicle merges with the flat target membrane. Hence, the operation of a fusion machine must be realized through the formation and expansion of a fusion pore. Delineating the relation between the fusion machine and the fusion pore thus emerges as a central goal in elucidating the mechanisms of membrane fusion. We summarize present knowledge of fusion machines and fusion pores studied in vitro, in neurons, and in neuroendocrine cells, and synthesize this knowledge into some specific and detailed hypotheses for exocytosis. PMID- 16689632 TI - RNA folding during transcription. AB - The evolution of RNA sequence needs to satisfy three requirements: folding, structure, and function. Studies on folding during transcription are related directly to folding in the cell. Understanding RNA folding during transcription requires the elucidation of structure formation and structural changes of the RNA, and the consideration of intrinsic properties of the RNA polymerase and other proteins that interact with the RNA. This review summarizes the research progress in this area and outlines the enormous challenges facing this field. Significant advancement requires the development of new experimental methods and theoretical considerations in all aspects of transcription and RNA folding. PMID- 16689633 TI - Roles of bilayer material properties in function and distribution of membrane proteins. AB - Structural, compositional, and material (elastic) properties of lipid bilayers exert strong influences on the interactions of water-soluble proteins and peptides with membranes, the distribution of transmembrane proteins in the plane of the membrane, and the function of specific membrane channels. Theoretical and experimental studies show that the binding of either cytoplasmic proteins or extracellular peptides to membranes is regulated by the presence of charged lipids and that the sorting of transmembrane proteins into or out of membrane microdomains (rafts) depends on several factors, including bilayer material properties governed by the presence of cholesterol. Recent studies have also shown that bilayer material properties modify the permeability of membrane pores, formed either by protein channels or by cell-lytic peptides. PMID- 16689634 TI - Electron tomography of membrane-bound cellular organelles. AB - Electron microscope tomography produces three-dimensional reconstructions and has been used to image organelles both isolated and in situ, providing new insight into their structure and function. It is analogous to the various tomographies used in medical imaging. Compared with light microscopy, electron tomography offers an improvement in resolution of 30- to 80-fold and currently ranges from 3 to 8 nm, thus filling the gap between high-resolution structure determinations of isolated macromolecules and larger-scale studies on cells and tissues by light microscopy. Here, we provide an introduction to electron tomography and applications of the method in characterizing organelle architecture that also show its power for suggesting functional significance. Further improvements in labeling modalities, imaging tools, specimen preparation, and reconstruction algorithms promise to increase the quality and breadth of reconstructions by electron tomography and eventually to allow the mapping of the cellular proteomes onto detailed three-dimensional models of cellular structure. PMID- 16689635 TI - Expanding the genetic code. AB - Recently, a general method was developed that makes it possible to genetically encode unnatural amino acids with diverse physical, chemical, or biological properties in Escherichia coli, yeast, and mammalian cells. More than 30 unnatural amino acids have been incorporated into proteins with high fidelity and efficiency by means of a unique codon and corresponding tRNA/aminoacyl-tRNA synthetase pair. These include fluorescent, glycosylated, metal-ion-binding, and redox-active amino acids, as well as amino acids with unique chemical and photochemical reactivity. This methodology provides a powerful tool both for exploring protein structure and function in vitro and in vivo and for generating proteins with new or enhanced properties. PMID- 16689636 TI - Radiolytic protein footprinting with mass spectrometry to probe the structure of macromolecular complexes. AB - Structural proteomics approaches using mass spectrometry are increasingly used in biology to examine the composition and structure of macromolecules. Hydroxyl radical-mediated protein footprinting using mass spectrometry has recently been developed to define structure, assembly, and conformational changes of macromolecules in solution based on measurements of reactivity of amino acid side chain groups with covalent modification reagents. Accurate measurements of side chain reactivity are achieved using quantitative liquid-chromatography-coupled mass spectrometry, whereas the side chain modification sites are identified using tandem mass spectrometry. In addition, the use of footprinting data in conjunction with computational modeling approaches is a powerful new method for testing and refining structural models of macromolecules and their complexes. In this review, we discuss the basic chemistry of hydroxyl radical reactions with peptides and proteins, highlight various approaches to map protein structure using radical oxidation methods, and describe state-of-the-art approaches to combine computational and footprinting data. PMID- 16689638 TI - Ribosome dynamics: insights from atomic structure modeling into cryo-electron microscopy maps. AB - Single-particle cryo-electron microscopy (cryo-EM) is the method of choice for studying the dynamics of macromolecular machines both at a phenomenological and, increasingly, at the molecular level, with the advent of high-resolution component X-ray structures and of progressively improving fitting algorithms. Cryo-EM has shed light on the structure of the ribosome during the four steps of translation: initiation, elongation, termination, and recycling. Interpretation of cryo-EM reconstructions of the ribosome in quasi-atomic detail reveals a picture in which the ribosome uses RNA not only to catalyze chemical reactions, but also as a means for signal transduction over large distances. PMID- 16689637 TI - The ESCRT complexes: structure and mechanism of a membrane-trafficking network. AB - The ESCRT complexes and associated proteins comprise a major pathway for the lysosomal degradation of transmembrane proteins and are critical for receptor downregulation, budding of the HIV virus, and other normal and pathological cell processes. The ESCRT system is conserved from yeast to humans. The ESCRT complexes form a network that recruits monoubiquitinated proteins and drives their internalization into lumenal vesicles within a type of endosome known as a multivesicular body. The structures and interactions of many of the components have been determined over the past three years, revealing mechanisms for membrane and cargo recruitment and for complex assembly. PMID- 16689639 TI - NMR techniques for very large proteins and rnas in solution. AB - Three-dimensional structure determination of small proteins and oligonucleotides by solution NMR is established. With the development of novel NMR and labeling techniques, structure determination is now feasible for proteins with a molecular mass of up to approximately 100 kDa and RNAs of up to 35 kDa. Beyond these molecular masses special techniques and approaches are required for applying NMR as a multiprobe method for structural investigations of proteins and RNAs. It is the aim of this review to summarize the NMR techniques and approaches available to advance the molecular mass limit of NMR both for proteins (up to 1 MDa) and RNAs (up to 100 kDa). Physical pictures of the novel techniques, their experimental applications, as well as labeling and assignment strategies are discussed and accompanied by future perspectives. PMID- 16689640 TI - Single-molecule analysis of RNA polymerase transcription. AB - The kinetics and mechanisms of transcription are now being investigated by a repertoire of single-molecule techniques, including optical and magnetic tweezers, high-sensitivity fluorescence techniques, and atomic force microscopy. Single-molecule techniques complement traditional biochemical and crystallographic approaches, are capable of detecting the motions and dynamics of individual RNAP molecules and transcription complexes in real time, and make it possible to directly measure RNAP binding to and unwinding of template DNA, as well as RNAP translocation along the DNA during transcript synthesis. PMID- 16689641 TI - Quantitative fluorescent speckle microscopy of cytoskeleton dynamics. AB - Fluorescent speckle microscopy (FSM) is a technology used to analyze the dynamics of macromolecular assemblies in vivo and in vitro. Speckle formation by random association of fluorophores with a macromolecular structure was originally discovered for microtubules. Since then FSM has been expanded to study other cytoskeleton and cytoskeleton-binding proteins. Specialized software has been developed to convert the stochastic speckle image signal into spatiotemporal maps of polymer transport and turnover in living cells. These maps serve as a unique quantitative readout of the dynamic steady state of the cytoskeleton and its responses to molecular and genetic interventions, allowing a systematic study of the mechanisms of cytoskeleton regulation and its effect on cell function. Here, we explain the principles of FSM imaging and signal analysis, outline the biological questions and corresponding methodological advances that have led to the current state of FSM, and give a glimpse of new FSM modalities under development. PMID- 16689642 TI - Water mediation in protein folding and molecular recognition. AB - Water is essential for life in many ways, and without it biomolecules might no longer truly be biomolecules. In particular, water is important to the structure, stability, dynamics, and function of biological macromolecules. In protein folding, water mediates the collapse of the chain and the search for the native topology through a funneled energy landscape. Water actively participates in molecular recognition by mediating the interactions between binding partners and contributes to either enthalpic or entropic stabilization. Accordingly, water must be included in recognition and structure prediction codes to capture specificity. Thus water should not be treated as an inert environment, but rather as an integral and active component of biomolecular systems, where it has both dynamic and structural roles. Focusing on water sheds light on the physics and function of biological machinery and self-assembly and may advance our understanding of the natural design of proteins and nucleic acids. PMID- 16689643 TI - Continuous membrane-cytoskeleton adhesion requires continuous accommodation to lipid and cytoskeleton dynamics. AB - The plasma membrane of most animal cells conforms to the cytoskeleton and only occasionally separates to form blebs. Previous studies indicated that many weak interactions between cytoskeleton and the lipid bilayer kept the surfaces together to counteract the normal outward pressure of cytoplasm. Either the loss of adhesion strength or the formation of gaps in the cytoskeleton enables the pressure to form blebs. Membrane-associated cytoskeleton proteins, such as spectrin and filamin, can control the movement and aggregation of membrane proteins and lipids, e.g., phosphoinositol phospholipids (PIPs), as well as blebbing. At the same time, lipids (particularly PIPs) and membrane proteins affect cytoskeleton and signaling dynamics. We consider here the roles of the major phosphatidylinositol-4,5-diphosphate (PIP2) binding protein, MARCKS, and PIP2 levels in controlling cytoskeleton dynamics. Further understanding of dynamics will provide important clues about how membrane-cytoskeleton adhesion rapidly adjusts to cytoskeleton and membrane dynamics. PMID- 16689644 TI - Cryo-electron microscopy of spliceosomal components. AB - Splicing is an essential step of gene expression in which introns are removed from pre-mRNA to generate mature mRNA that can be translated by the ribosome. This reaction is catalyzed by a large and dynamic macromolecular RNP complex called the spliceosome. The spliceosome is formed by the stepwise integration of five snRNPs composed of U1, U2, U4, U5, and U6 snRNAs and more than 150 proteins binding sequentially to pre-mRNA. To study the structure of this particularly dynamic RNP machine that undergoes many changes in composition and conformation, single-particle cryo-electron microscopy (cryo-EM) is currently the method of choice. In this review, we present the results of these cryo-EM studies along with some new perspectives on structural and functional aspects of splicing, and we outline the perspectives and limitations of the cryo-EM technique in obtaining structural information about macromolecular complexes, such as the spliceosome, involved in splicing. PMID- 16689645 TI - Mechanotransduction involving multimodular proteins: converting force into biochemical signals. AB - Cells can sense and transduce a broad range of mechanical forces into distinct sets of biochemical signals that ultimately regulate cellular processes, including adhesion, proliferation, differentiation, and apoptosis. Deciphering at the nanoscale the design principles by which sensory elements are integrated into structural protein motifs whose conformations can be switched mechanically is crucial to understand the process of transduction of force into biochemical signals that are then integrated to regulate mechanoresponsive pathways. While the major focus in the search for mechanosensory units has been on membrane proteins such as ion channels, integrins, and associated cytoplasmic complexes, a multimodular design of tandem repeats of various structural motifs is ubiquitously found among extracellular matrix proteins, as well as cell adhesion molecules, and among many intracellular players that physically link transmembrane proteins to the contractile cytoskeleton. Single-molecule studies have revealed an unexpected richness of mechanosensory motifs, including force regulated conformational changes of loop-exposed molecular recognition sites, intermediate states in the unraveling pathway that might either expose cryptic binding or phosphorylation sites, or regions that display enzymatic activity only when unmasked by force. Insights into mechanochemical signal conversion principles will also affect various technological fields, from biotechnology to tissue engineering and drug development. PMID- 16689646 TI - Geothermal synthesis and metabolism. PMID- 16689647 TI - Fossil DNA in cretaceous black shales: myth or reality? PMID- 16689648 TI - Molecular signals from ancient materials: challenges to deep-biosphere and paleo environmental research--a response to the comments of Sinninghe Damste and Coolen. PMID- 16689649 TI - Astrobiologically interesting stars within 10 parsecs of the sun. AB - The existence of life based on carbon chemistry and water oceans relies upon planetary properties, chiefly climate stability, and stellar properties, such as mass, age, metallicity, and galactic orbits. The latter can be well constrained with present knowledge. We present a detailed, up-to-date compilation of the atmospheric parameters, chemical composition, multiplicity, and degree of chromospheric activity for the astrobiologically interesting solar-type stars within 10 parsecs of the Sun. We determined their state of evolution, masses, ages, and space velocities, and produced an optimized list of candidates that merit serious scientific consideration by the future space-based interferometry probes aimed at directly detecting Earthsized extrasolar planets and seeking spectroscopic infrared biomarkers as evidence of photosynthetic life. The initially selected stars number 33 solar-type within the total population (excluding some incompleteness for late M-dwarfs) of 182 stars closer than 10 parsecs. A comprehensive and detailed data compilation for these objects is still lacking; a considerable amount of recent data has so far gone unexplored in this context. We present 13 objects as the nearest "biostars," after eliminating multiple stars, young, chromospherically active, hard x-ray- emitting stars, and low metallicity objects. Three of these "biostars"-- Zeta Tucanae, Beta Canum Venaticorum, and 61 Virginis -- closely reproduce most of the solar properties and are considered as premier targets. We show that approximately 7% of the nearby stars are optimally interesting targets for exobiology. PMID- 16689650 TI - Microbial survival rates of Escherichia coli and Deinococcus radiodurans under low temperature, low pressure, and UV-Irradiation conditions, and their relevance to possible Martian life. AB - Viability rates were determined for microbial populations of Escherichia coli and Deinococcus radiodurans under the environmental stresses of low temperature (-35 degrees C), low-pressure conditions (83.3 kPa), and ultraviolet (UV) irradiation (37 W/m(2)). During the stress tests the organisms were suspended in saltwater soil and freshwater soil media, at variable burial depths, and in seawater. Microbial populations of both organisms were most susceptible to dehydration stress associated with low-pressure conditions, and to UV irradiation. However, suspension in a liquid water medium and burial at larger depths (5 cm) improved survival rates markedly. Our results indicate that planetary surfaces that possess little to no atmosphere and have low water availability do not constitute a favorable environment for terrestrial microorganisms. PMID- 16689651 TI - Impact craters as biospheric microenvironments, Lawn Hill Structure, Northern Australia. AB - Impact craters on Mars act as traps for eolian sediment and in the past may have provided suitable microenvironments that could have supported and preserved a stressed biosphere. If this is so, terrestrial impact structures such as the 18 km-diameter Lawn Hill Structure, in northern Australia, may prove useful as martian analogs. We sampled outcrop and drill core from the carbonate fill of the Lawn Hill Structure and recorded its gamma-log signature. Facies data along with whole rock geochemistry and stable isotope signatures show that the crater fill is an outlier of the Georgina Basin and was formed by impact at, or shortly before, approximately 509-506 million years ago. Subsequently, it was rapidly engulfed by the Middle Cambrian marine transgression, which filled it with shallow marine carbonates and evaporites. The crater formed a protected but restricted microenvironment in which sediments four times the thickness of the nearby basinal succession accumulated. Similar structures, common on the martian surface, may well have acted as biospheric refuges as the planet's water resources declined. Low-pH aqueous environments on Earth similar to those on Mars, while extreme, support diverse ecologies. The architecture of the eolian crater fill would have been defined by long-term ground water cycles resulting from intermittent precipitation in an extremely arid climate. Nutrient recycling, critical to a closed lacustrine sub-ice biosphere, could be provided by eolian transport onto the frozen water surface. PMID- 16689652 TI - Serpentinization and its implications for life on the early Earth and Mars. AB - Ophiolites, sections of ocean crust tectonically displaced onto land, offer significant potential to support chemolithoautotrophic life through the provision of energy and reducing power during aqueous alteration of their highly reduced mineralogies. There is substantial chemical disequilibrium between the primary olivine and pyroxene mineralogy of these ophiolites and the fluids circulating through them. This disequilibrium represents a potential source of chemical energy that could sustain life. Moreover, E (h)-pH conditions resulting from rock water interactions in ultrabasic rocks are conducive to important abiotic processes antecedent to the origin of life. Serpentinization--the reaction of olivine- and pyroxene-rich rocks with water--produces magnetite, hydroxide, and serpentine minerals, and liberates molecular hydrogen, a source of energy and electrons that can be readily utilized by a broad array of chemosynthetic organisms. These systems are viewed as important analogs for potential early ecosystems on both Earth and Mars, where highly reducing mineralogy was likely widespread in an undifferentiated crust. Secondary phases precipitated during serpentinization have the capability to preserve organic or mineral biosignatures. We describe the petrology and mineral chemistry of an ophiolite hosted cold spring in northern California and propose criteria to aid in the identification of serpentinizing terranes on Mars that have the potential to harbor chemosynthetic life. PMID- 16689653 TI - Martian CH(4): sources, flux, and detection. AB - Recent observations have detected trace amounts of CH(4) heterogeneously distributed in the martian atmosphere, which indicated a subsurface CH(4) flux of ~2 x 10(5) to 2 x 10(9) cm(2) s(1). Four different origins for this CH(4) were considered: (1) volcanogenic; (2) sublimation of hydrate- rich ice; (3) diffusive transport through hydrate-saturated cryosphere; and (4) microbial CH(4) generation above the cryosphere. A diffusive flux model of the martian crust for He, H(2), and CH(4) was developed based upon measurements of deep fracture water samples from South Africa. This model distinguishes between abiogenic and microbial CH(4) sources based upon their isotopic composition, and couples microbial CH(4) production to H(2) generation by H(2)O radiolysis. For a He flux of approximately 10(5) cm(2) s(1) this model yields an abiogenic CH(4) flux and a microbial CH(4) flux of approximately 10(6) and approximately 10(9) cm(2) s(1), respectively. This flux will only reach the martian surface if CH(4) hydrate is saturated in the cryosphere; otherwise it will be captured within the cryosphere. The sublimation of a hydrate-rich cryosphere could generate the observed CH(4) flux, whereas microbial CH(4) production in a hypersaline environment above the hydrate stability zone only seems capable of supplying approximately 10(5) cm(2) s(1) of CH(4). The model predicts that He/H(2)/CH(4)/C(2)H(6) abundances and the C and H isotopic values of CH(4) and the C isotopic composition of C(2)H(6) could reveal the different sources. Cavity ring-down spectrometers represent the instrument type that would be most capable of performing the C and H measurements of CH(4) on near future rover missions and pinpointing the cause and source of the CH(4) emissions. PMID- 16689655 TI - Rabies viral mechanisms to escape the IFN system: the viral protein P interferes with IRF-3, Stat1, and PML nuclear bodies. AB - Interferons (IFNs) are a family of secreted proteins with antiviral, antiproliferative, and immunomodulatory activities. The different biologic actions of IFN are believed to be mediated by the products of specifically IFN stimulated genes (ISG) in the target cells. The IFN response is the first line of defense against viral infections. Viruses, which require the cellular machinery for their replication, have evolved different ways to counteract the action of IFN by inhibiting IFN production or Jak-Stat signaling or by altering ISG products. This review focuses on the role of viral proteins from the RNA virus family, particularly rabies P protein. P protein mediates inhibition of the IFN system by different pathways: it inhibits IFN production by impairing IFN regulatory factor-3 (IRF-3) phosphorylation and IFN signaling by blocking nuclear transport of Stat1 and alters promyelocytic leukemia (PML) nuclear bodies by retaining PML in the cytoplasm. PMID- 16689656 TI - Proteasome-mediated proteolysis of the interleukin-10 receptor is important for signal downregulation. AB - The cytokine interleukin-10 (IL-10) is an important regulator of immune cell function, proliferation, and survival. The IL-10 receptor (IL-10R) consists of two subunits, IL-10R1 and IL-10R2, both belonging to the class II cytokine receptor superfamily. Like other members of the cytokine receptor superfamily, IL 10R stimulation leads to activation of Jak family kinases and Stat transcription factors. To identify additional signal transduction pathways used by the IL-10R, we purified 92-kDa and 100-kDa proteins that coprecipitated with IL-10R1 from IL 10-stimulated cells. Both proteins were found to be related to the 97-kDa subunit of the regulatory component of the 26S proteasome. Subsequent studies confirmed that the IL-10R1 undergoes ligand- dependent internalization and proteasome mediated degradation. An IL-10R1 cytoplasmic domain mutant deficient for internalization exhibited prolonged signaling through Jak1 and Stat3, reinforcing the importance of receptor internalization for signal termination. PMID- 16689657 TI - The neutrophil/Th1 lymphocyte balance and the therapeutic effect of granulocyte colony-stimulating factor in TNBS-induced colitis of rat strains. AB - Intramucosal neutrophil infiltration is related to the activity of ulcerative colitis, and Th1 immunity is responsible for the onset of Crohn's disease. We examined the therapeutic effects of recombinant human granulocyte colony stimulating factor (rHuG-CSF) in the two types of 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis of five rat strains. SD and DA rats showed much lower mRNA expression levels of endogenous G-CSF in lipopolysaccharide (LPS)-stimulated splenocytes than did Lewis, F344, and BN rats. On day 7 after anal instillation of TNBS, SD and DA rats demonstrated massive lymphocyte infiltration with an interferon-gamma (IFN-gamma) mRNA upregulation, whereas Lewis, F344, and BN rats showed an intense submucosal neutrophil accumulation with high tumor necrosis factor-alpha (TNF-alpha) mRNA levels. A 5-day course of rHuG-CSF pretreatment (250 microg/kg/day, s.c.) reduced the elevated levels of both cytokines. The treatment improved the survival rate of DA and reduced the degree of body weight loss of SD, while not significantly influencing the wasting disease of other strains. Interleukin-10 (IL-10) mRNA levels were highly upregulated by rHuG-CSF treatment on day 1 in the neutrophil-dominant lesions of F344 but not in the Th1 type lesions of SD, and IL-12p35 mRNA levels were downregulated in both. A supply of G-CSF prevents the onset of Th1-type TNBS colitis and does not deteriorate neutrophil-dominant chronic colitis in hosts showing higher expression of endogenous G-CSF. PMID- 16689658 TI - Antifibrogenic effect of IFN-alpha2b on hepatic stellate cell activation by human hepatocytes. AB - Liver fibrosis commonly occurs in patients with hepatitis C virus (HCV) infection as a consequence of the chronic liver damage, thus leading to the development of liver cirrhosis. When hepatic stellate cells (HSCs) become active, they play an essential role in liver fibrogenesis. In this study, interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha), commonly elevated in chronic C hepatitis, stimulate the production of matrix metalloprotease-9 (MMP-9) by human hepatocytes at a transcriptional and translational level, but the addition of recombinant interferon-alpha2b (rIFN-alpha2b) hampers this effect. Furthermore, a human HSC line is activated in vitro by incubation with human MMP-9 in the presence of collagen I, and this effect is blocked by the MMP inhibitor BB94. A similar activation was observed when incubating HSCs with conditioned medium of hepatocytes previously stimulated with IL-1beta and TNF-alpha but not when using conditioned medium of hepatocytes costimulated with IL-1beta or TNF-alpha together with rIFN-alpha2b. In conclusion, our results show that hepatocytes stimulated by inflammatory cytokines participate in the activation of HSCs via MMP-9 production and that antiviral therapy modulates such activation. PMID- 16689659 TI - SARS-CoV virus-host interactions and comparative etiologies of acute respiratory distress syndrome as determined by transcriptional and cytokine profiling of formalin-fixed paraffin-embedded tissues. AB - These studies attempt to understand more fully the host response and pathogenesis associated with severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) by monitoring gene expression using formalin-fixed paraffin-embedded (FFPE) pulmonary autopsy tissues. These tissues were from patients in different hospitals in Singapore who were diagnosed with various microbial infections, including SARS-CoV, that caused acute respiratory distress syndrome (ARDS). Global expression patterns showed limited correlation between end-stage ARDS and the initiating pathogen, but when focusing on a subset of genes implicated in pulmonary pathogenesis, molecular signatures of pulmonary disease were obtained and appeared to be influenced by preexisting pulmonary complications and also bacterial components of infection. Many factors detected during pulmonary damage and repair, such as extracellular matrix (ECM) components, transforming growth factor (TGF) enhancers, acute-phase proteins, and antioxidants, were included in the molecular profiles of these ARDS lung tissues. In addition, differential expression of cytokines within these pulmonary tissues were observed, including notable genes involved in the interferon (IFN) pathway, such as Stat1, IFN regulatory factor-1 (IRF-1), interleukin-6 (IL-6), IL-8, and IL-18, that are often characterized as elevated in ARDS patients. PMID- 16689660 TI - Lung epithelial NF-kappaB and Stat1 signaling in response to CD8+ T cell antigen recognition. AB - CD8+ T cell recognition of viral antigens presented by lung epithelial cells is important in the clearance of respiratory viral infection but may cause considerable injury to the lung. We have shown that a critical event of this type of injury is the activation of target epithelial cells and expression of chemokines by these cells. In this study, epithelial gene expression and transcription factor activation triggered by specific CD8+ T cell antigen recognition was examined in vitro and in vivo. T cell recognition triggers expression profiles of tumor necrosis factor-alpha (TNF-alpha)-dependent and interferon-gamma (IFN-gamma)-dependent genes in epithelial target cells. Consistent with these profiles, transcription factors nuclear factor-kappaB (NF kappaB) and activator protein-1 (AP-1) were activated in lung epithelial cells of wild-type (WT) mice but not TNF receptor 1 (TNFR1)-deficient mice after CD8+ T cell recognition in vivo. In contrast, Stat1 activation and Stat1-dependent genes, such as IFN regulatory factor-1 (IRF-1) and guanylate-binding protein-2 (GBP-2), were induced to a similar extent in epithelial cells of both WT and TNFR1-deficient mice, indicating that this pathway is insufficient to induce pulmonary immunopathology in the absence of NF-kappaB-dependent transcriptional activation. Antibody neutralization of TNF-alpha abrogated epithelial monocyte chemotactic protein-1 (MCP-1) and macrophage inflammatory protein-2 (MIP-2) production in vitro as well as pulmonary immunopathology in vivo, confirming the primary importance of this cytokine in CD8+ T cell-mediated immunopathology. PMID- 16689661 TI - In silico genomic analysis of the human and murine guanylate-binding protein (GBP) gene clusters. AB - The guanylate-binding proteins (GBPs) were among the first interferon (IFN) stimulated genes (ISGs) discovered, but until recently, little was known about their functions and even less about the composition of the gene family. Analysis of the promoter of human GBP-1 contributed significantly toward the understanding of Jak-Stat signaling and the delineation of the IFN-gamma activation site (GAS) and IFN-stimulated response element (ISRE) promoter elements. In this study, we have examined the genomic arrangement and composition of the GBPs in both mouse and humans. There are seven GBP paralogs in humans and at least one pseudogene, all of which are located in a cluster of genes on chromosome 1. Five of the six MuGBPs and a GBP pseudogene are clustered in a syntenic region on chromosome 3. The sixth MuGBP, MuGBP-4, and three GBP pseudogenes are located on chromosome 5. As might be expected, the GBPs share similar genomic organizations of introns and exons. Five of the MuGBPs had previously been shown to be coordinately induced by IFNs, and as expected, all of the MuGBPs have GAS and ISRE elements in their promoters. Interestingly, not all of the HuGBPs have GAS and ISRE elements, suggesting that not all GBPs are IFN responsive in humans. PMID- 16689662 TI - IFN-alpha1,8 inhibits tumor-induced angiogenesis in murine angiosarcomas. AB - Interferon-alpha (IFN-alpha) has proved effective in the treatment of hemangiomas, hemangioblastomas, and Kaposi's sarcoma. To investigate the ability of IFNs to inhibit angiosarcoma, we used two transformed murine endothelial cell lines that form angiosarcomas in vivo. SVR and MS1-VEGF cell lines express oncogenic H-ras or vascular endothelial growth factor (VEGF), respectively. IFN alpha1,8, which is active against murine and human cells, inhibited SVR and MS1 VEGF proliferation in vitro by 40% at 10(3) U/mL (p = 0.028). In vivo, IFN alpha1,8 inhibited SVR tumor volume by 71% (p = 0.047) and MS1-VEGF volume by 79% (p = 0.003). Tumor-induced angiogenesis was decreased in SVR tumors by 52% (p = 0.005) and in MS1-VEGF tumors by 58% (p = 0.001). Sera from IFN-alpha1,8-treated mice bearing either SVR or MS1-VEGF tumors demonstrated a 5-fold increase in IP 10/CXCL10 (p = 0.001), an IFN-induced antiangiogenic protein. Both recombinant IP 10 and IFN-alpha1,8 inhibited human umbilical vein endothelial cell (HUVEC) vessel formation in the fibrin gel assay, a three-dimensional culture model of angiogenesis, by 56% at 25 ng/mL and 50% at 1.2 ng/mL, respectively (p < 0.001). An IP-10 blocking antibody restored vessel formation to 80% of untreated controls (p = 0.001). Given the magnitude of the in vivo response, these data suggested that the antitumor effects of IFN-alpha1,8 were likely mediated through angiogenesis inhibition rather than solely by direct inhibition of tumor cell proliferation. PMID- 16689664 TI - Overexpression of SOD1 in transgenic rats attenuates nuclear translocation of endonuclease G and apoptosis after spinal cord injury. AB - Spinal motor neurons are selectively vulnerable after spinal cord injury (SCI). Recent studies suggest they undergo apoptosis after caspase activation through a mitochondria-dependent apoptosis pathway, and that oxidative stress after SCI is likely to play a role. However, other signaling pathways of apoptosis that involve mitochondria have not been thoroughly studied after SCI. Apoptosis inducing factor (AIF) and endonuclease G (EndoG) are mitochondrial apoptogenic proteins that are capable of inducing neuronal apoptosis when translocated from mitochondria to nuclei through a caspase-independent pathway. In this study, we examined translocation of these proteins and apoptotic cell death of motor neurons. The role of oxidative stress was also studied using transgenic (Tg) rats that overexpress the intrinsic antioxidant copper/zinc-superoxide dismutase (SOD1). Western blots and an activity assay demonstrated that a greater amount of SOD1 and higher activity of SOD presented in mitochondria of Tg rats compared with wild-type (Wt) rats. Immunohistochemistry and Western blots showed translocation of EndoG and AIF from mitochondria to nuclei in motor neurons 1 day after SCI in both groups of rats. However, there was significantly less translocation of EndoG in the Tg rats compared with the Wt rats. Less apoptotic cell death was detected in the Tg rats than in the Wt rats 3 days after SCI. These results suggest that translocation of EndoG and AIF from mitochondria to nuclei may initiate a caspase-independent pathway of apoptosis. An increased level of SOD1 in mitochondria conceivably reduces oxidative stress, thereby attenuating EndoG translocation, and resulting in reduction of caspase independent apoptosis. PMID- 16689665 TI - Inhibition of Fas-mediated apoptosis through administration of soluble Fas receptor improves functional outcome and reduces posttraumatic axonal degeneration after acute spinal cord injury. AB - Fas receptor activation has been implicated in inflammatory responses, programmed cell death, Wallerian degeneration in neural injury and the axotomy induced death of motoneurons. Recent work using transection models of spinal cord injury (SCI) demonstrated that neutralization of Fas ligand with antibodies may promote axonal regeneration and functional recovery. Moreover, recent studies from our laboratory in mutant mice with deficient expression of Fas, show reduced cell death and enhanced behavioural recovery after SCI. The present paper examines the effects of soluble Fas receptor (sFasR) administration on inhibition of Fas receptor-Fas ligand interaction in the setting of acute SCI in vitro and in vivo. An in vitro model of SCI demonstrated that sFasR administration decreases cell death as assessed by propidium iodide fluorescence. Furthermore, in a moderately severe in vivo clip compression model of SCI at C7-T1, we demonstrate that subarachnoid infusion of sFasR results in increased neuron and oligodendrocyte survival, improved tissue and long tract axonal preservation, reduced apoptotic cell death and enhanced functional neurological outcome after acute SCI. These results strongly suggest that inhibiting Fas receptor activation is neuroprotective after acute SCI and that this strategy may have important translational significance. PMID- 16689666 TI - Delayed intervention with transplants and neurotrophic factors supports recovery of forelimb function after cervical spinal cord injury in adult rats. AB - The adult central nervous system is capable of considerable anatomical reorganization and functional recovery after injury. Functional outcomes, however, vary greatly, depending upon size and location of injury, type and timing of intervention, and type of recovery and plasticity evaluated. The present study was undertaken to assess the recovery of skilled and unskilled forelimb function in adult rats after a C5/C6 spinal cord over-hemisection and delayed intervention with fetal spinal cord transplants and neurotrophins. Recovery of forelimb function was evaluated during both target reaching (a skilled behavior) and vertical exploration (an unskilled behavior). Anatomical tracing and immunohistochemistry were used to assess the growth of descending raphespinal, corticospinal, and rubrospinal fibers at the injury site, tracts that normally confer forelimb function. Delayed intervention with transplants and either brain-derived neurotrophic factor (BDNF) or neurotrophin-3 (NT-3) restored skilled left forelimb reaching to pre-injury levels. Animals showed recovery of normal reaching movements rather than compensation with abnormal movements. Transplants and NT-3 also improved right forelimb use during an unskilled vertical exploration, but not skilled right reaching. Intervention with fetal transplant tissue supported the growth of descending serotonergic, corticospinal, and rubrospinal fibers into the transplant at the lesion site. The addition of neurotrophins, however, did not significantly increase axonal growth at the lesion site. These studies suggest that the recovery of skilled and unskilled forelimb use is possible after a large cervical spinal cord injury following delayed intervention with fetal spinal cord and neurotrophins. Plasticity of both spared and axotomized descending pathways likely contributes to the functional recovery observed. PMID- 16689667 TI - Basso Mouse Scale for locomotion detects differences in recovery after spinal cord injury in five common mouse strains. AB - Genetically engineered mice are used extensively to examine molecular responses to spinal cord injury (SCI). Inherent strain differences may confound behavioral outcomes; therefore, behavioral characterization of several strains after SCI is warranted. The Basso, Beattie, Bresnahan Locomotor Rating Scale (BBB) for rats has been widely used for SCI mice, but may not accurately reflect their unique recovery pattern. This study's purpose was to develop a valid locomotor rating scale for mice and to identify strain differences in locomotor recovery after SCI. We examined C57BL/6, C57BL/10, B10.PL, BALB/c, and C57BL/6x129S6 F1 strains for 42 days after mild, moderate, and severe contusive SCI or transection of the mid thoracic spinal cord. Contusions were created using the Ohio State University electromagnetic SCI device which is a displacement-driven model, and the Infinite Horizon device, which is a force-driven model. Attributes and rankings for the Basso Mouse Scale for Locomotion (BMS) were determined from frequency analyses of seven locomotor categories. Mouse recovery differed from rats for coordination, paw position and trunk instability. Disagreement occurred across six expert raters using BBB (p < 0.05) but not BMS to assess the same mice. BMS detected significant differences in locomotor outcomes between severe contusion and transection (p < 0.05) and SCI severity gradations resulting from displacement variations of only 0.1 mm (p < 0.05). BMS demonstrated significant face, predictive and concurrent validity. Novice BMS raters with training scored within 0.5 points of experts and demonstrated high reliability (0.92-0.99). The BMS is a sensitive, valid and reliable locomotor measure in SCI mice. BMS revealed significantly higher recovery in C57BL/10, B10.PL and F1 than the C57BL/6 and BALB/c strains after moderate SCI (p < 0.05). The differing behavioral response to SCI suggests inherent genetic factors significantly impact locomotor recovery and must be considered in studies with inbred or genetically engineered mouse strains. PMID- 16689668 TI - Augmented locomotor recovery after spinal cord injury in the athymic nude rat. AB - The immune response contributes to ongoing secondary tissue destruction following spinal cord injury (SCI). Although infiltrating neutrophils and monocytes have been well studied in this process, T-cells have received less attention. The objective of this study was to assess locomotor recovery and tissue morphology after SCI in athymic (nude) rats, in which T-cell numbers are reduced. Results in athymic rats were compared with heterozygote littermates with normal T-cell profiles and with Sprague-Dawley rats from previous studies in our lab. Following transection of rat spinal cords at T10, we assessed the animals' locomotor recovery on a weekly basis for up to 11 weeks, using the Basso-Beattie-Bresnahan locomotor rating scale. Nude rats showed better locomotor recovery than did heterozygote or Sprague-Dawley rats, achieving scores of 5.6 +/- 0.8 versus 1.0 +/- 0.0, respectively (p = 0.002), at 4 weeks postinjury. The improved recovery of nude rats persisted for the 11-week postinjury assessment period, and was consistent with improved spinal reflexes rather than with recovery of descending motor pathways. Anatomical evaluation at 11 weeks indicated no difference in nude versus heterozygote rats in the size or distribution of cavities caudal to the transection site, but secondary damage was more severe rostral to the transection site in heterozygote rats. In neither group did cavities extend beyond 4 mm caudal to the transection site, and were therefore not directly responsible for the functional differences between the two groups. Cellular expression of the microglia/macrophage antigen ectodysplasin A (ED1) was reduced in nude rats as compared to heterozygotes, but no difference was observed in expression levels of 5-hydroxytryptamine, the 200-kDa neurofilament, or glial fibrillary acidic protein. The findings of the study show that a reduction in T-cell numbers significantly improves locomotor recovery after spinal cord transection, indicating a deleterious role for these immune cells in neural repair after trauma. PMID- 16689669 TI - Early morphologic and spectroscopic magnetic resonance in severe traumatic brain injuries can detect "invisible brain stem damage" and predict "vegetative states". AB - A precise evaluation of the brain damage in the first days of severe traumatic brain injured (TBI) patients is still uncertain despite numerous available cerebral evaluation methods and imaging. In 5-10% of severe TBI patients, clinicians remain concerned with prolonged coma and long-term marked cognitive impairment unexplained by normal morphological T2 star, flair, and diffusion magnetic resonance imaging (MRI). For this reason, we prospectively assessed the potential value of magnetic resonance spectroscopy (MRS) of the brain stem to evaluate the functionality of the consciousness areas. Forty consecutive patients with severe TBI were included. Single voxel proton MRS of the brain stem and morphological MRI of the whole brain were performed at day 17.5 +/- 6.4. Disability Rating Scale and Glasgow Outcome Scale (GOS) were evaluated at 18 months posttrauma. MRS appeared to be a reliable tool in the exploration of brainstem metabolism in TBI. Three different spectra were observed (normal, cholinergic reaction, or neuronal damage) allowing an evaluation of functional damage. MRS disturbances were not correlated with anatomical MRI lesions suggesting that the two techniques are strongly complementarity. In two GOS 2 vegetative patients with normal morphological MRI, MRS detected severe functional damage of the brainstem (NAA/Cr < 1.50) that was described as "invisible brain stem damage." MRI and MRS taken separately could not distinguish patients GOS 3 (n = 7) from GOS 1-2 (n = 11) and GOS 4-5 (n = 20). However, a principal component analysis of combined MRI and MRS data enabled a clear-cut separation between GOS 1-2, GOS 3, and GOS 4-5 patients with no overlap between groups. This study showed that combined MRI and MRS provide a reliable evaluation of patients presenting in deep coma, specially when there are insufficient MRI lesions of the consciousness pathways to explain their status. In the first few days post-trauma metabolic (brainstem spectroscopy) and morphological (T2 star and Flair) MRI studies can predict the long-term neurological outcome, especially the persistent vegetative states and minimally conscious state. PMID- 16689670 TI - Postinjury administration of pituitary adenylate cyclase activating polypeptide (PACAP) attenuates traumatically induced axonal injury in rats. AB - Pituitary adenylate cyclase activating polypeptide (PACAP) has several different actions in the nervous system. Numerous studies have shown its neuroprotective effects both in vitro and in vivo. Previously, it has been demonstrated that PACAP reduces brain damage in rat models of global and focal cerebral ischemia. Based on the protective effects of PACAP in cerebral ischemia and the presence of common pathogenic mechanisms in cerebral ischemia and traumatic brain injury (TBI), the aim of the present study was to investigate the possible protective effect of PACAP administered 30 min or 1 h postinjury in a rat model of diffuse axonal injury. Adult Wistar male rats were subjected to impact acceleration, and PACAP was administered intracerebroventricularly 30 min (n = 4), and 1 h after the injury (n = 5). Control animals received the same volume of vehicle at both time-points (n = 5). Two hours after the injury, brains were processed for immunohistochemical localization of damaged axonal profiles displaying either beta-amyloid precursor protein (beta-APP) or RMO-14 immunoreactivity, both considered markers of specific features of traumatic axonal injury. Our results show that treatment with PACAP (100 microg) 30 min or 1 h after the induction of TBI resulted in a significant reduction of the density of beta-APP-immunopositive axon profiles in the corticospinal tract (CSpT). There was no significant difference between the density of beta-APP-immunopositive axons in the medial longitudinal fascicle (MLF). PACAP treatment did not result in significantly different number of RMO-14-immunopositive axonal profiles in either brain areas 2 hours post-injury compared to normal animals. While the results of this study highlighted the complexity of the pathogenesis and manifestation of diffuse axonal injury, they also indicate that PACAP should be considered a potential therapeutic agent in TBI. PMID- 16689671 TI - Autoradiographic analysis of mouse brain kinin B1 and B2 receptors after closed head trauma and ability of Anatibant mesylate to cross the blood-brain barrier. AB - The potent non-peptide B2 receptor (R) antagonist, Anatibant mesylate (Ms) (LF 16 0687 Ms), reduces brain edema and improves neurological function recovery in various focal and diffuse models of traumatic brain injury in rodents. In the present study, alteration of kinin B1 and B2R after closed head trauma (CHT) and in vivo binding properties of Anatibant Ms (3 mg/kg, s.c.) injected 30 min after CHT were studied in mice by autoradiography using the radioligands [125I]HPP-Hoe 140 (B2R), and [125I]HPP-des-Arg10-Hoe 140 (B1R). Whereas B1R is barely detected in most brain regions, B2R is extensively distributed, displaying the highest densities in the hindbrain. CHT was associated with a slight increase of B1R and a decrease of B2R (10-50%) in several brain regions. Anatibant Ms (Ki = 22 pM) displaced the B2R radioligand from its binding sites in several areas of the forebrain, basal ganglia and hindbrain. Displacement was achieved in 1 h and persisted at 4 h post-injection. The inhibition did not exceed 50% of the total specific binding in non-injured mice. After CHT, the displacement by Anatibant Ms was higher and almost complete in the cortex, caudate putamen, thalamus, hippocampus, medial geniculate nucleus, ventral tegmental area, and raphe. Evans blue extravasation in brain tissue at 4 h after CHT was abolished by Anatibant Ms. It appeared that Anatibant Ms penetrated into the brain in sufficient amounts, particularly after disruption of the blood-brain barrier, to account for its B2R-mediated neuro- and vascular protective effects. The diminished binding of B2R after CHT may reflect the occupancy or internalization of B2R following the endogenous production of bradykinin (BK). PMID- 16689672 TI - Reduced collagen degradation in polytraumas with traumatic brain injury causes enhanced osteogenesis. AB - Patients with traumatic brain injury (TBI) and skeletal injuries have increased rates of excessive bone healing (EH = hypertrophic callus formation and/or heterotopic ossification). Polytrauma patients are often attributed higher rates of delayed fracture union. This study compares 182 total fractures in 29 isolated polytrauma patients (POLY) and 48 patients after TBI and polytrauma (TBI+POLY), examining the clinical parameters of EH versus delay. A subset of 28 patients (13 TBI+POLY, 15 POLY) underwent serological testing for the following bone turnover parameters: carboxy-terminal extension peptide of type 1 procollagen (P1CP), pyridinolene cross-linked carboxy-terminal telopeptide (1CTP), insulin-like growth factor-1 (IGF-1), insulin-like growth factor binding protein-3 (IGFBP-3), and basic fibroblast growth factor (bFGF). There were higher rates of delayed union in the POLY patients (45% vs. 23%) and EH in the TBI+POLY patients (33% vs. 17%) (not significant = NS). More delayed unions were observed in diaphyseal fractures suffered by POLY (28%) than in TBI+POLY (15%) patients (NS). EH after pelvic fracture was apparent in 52% TBI+POLY and in 21% POLY fractures (NS). P1CP levels did not differ between the groups, but the collagen breakdown parameter 1CTP was significantly higher in the POLY group (p = 0.01-0.04). IGF-1 levels were below normal in both groups, and did not differ. IGFBP-3, an IGF-1 inhibiting and collagenase-3-activating protein, was significantly higher in POLY patients (p = 0.017-0.037). bFGF levels did not vary between groups. Increased serum levels of 1CTP and IGFBP-3 in POLY patients suggest that EH in TBI patients is secondary to decreased collagen breakdown rather than increased synthesis. PMID- 16689673 TI - Mass-related traumatic tissue displacement and behavior: a screen for treatments that reduce [corrected] harm to bystander cells and recovery of function. AB - In this study, we focused on a preclinical model of brain compression injury that has relevance to pathological conditions such as tumor, hematoma, blood clot, and intracerebral bony fragment. We investigated behavioral impairment as a result of rapid-onset small mass, and the factors involved in lesion formation and neuroplasticity. An epidural bead implantation method was adopted. Two sizes (1.5 mm and 2.0 mm thick) of hemisphere-shaped beads were used. The beads were implanted into various locations over the sensorimotor cortex (SMC--anterior, middle and posterior). The effects of early versus delayed bead removal were examined to model clinical neurosurgical or other treatment procedures. Forelimb and hind-limb behavioral deficits and recovery were observed, and histological changes were quantified to determine brain reaction to focal compression. Our results showed that the behavioral deficits of compression were influenced by the location, timing of compression release, and magnitude of compression. Even persistent compression by the thicker bead (2.0 mm) caused only minor behavioral deficits, followed by fast recovery within a week in most animals, suggesting a mild lesion pattern for this model. Brain tissue was compressed into a deformed shape under pressure with slight tissue damage, evidenced by pathological evaluation on hematoxylin and eosin (H&E)- and TUNEL-stained sections. Detectable but not severe behavioral dysfunction exhibited by this model makes it particularly suitable for direct assessment of adverse effects of interventions on neuroplasticity after brain compression injury. This model may permit development of treatment strategies to alleviate brain mass effects, without disrupting neuroplasticity. PMID- 16689679 TI - Characterization of a uricase from Bacillus fastidious A.T.C.C. 26904 and its application to serum uric acid assay by a patented kinetic uricase method. AB - An intracellular uricase from Bacillus fastidious A.T.C.C. 26904 was characterized and evaluated for serum uric acid assay by a patented kinetic uricase method. The active uricase was 151 kDa by gel filtration through Sephadex G-200. Both SDS/PAGE and matrix-assisted laser-desorption ionization-time-of flight MS resolved a single polypeptide with a molecular mass of approx. 36.0 kDa. The N-terminal sequence was AERTMFYGKGDV. The optimum pH for this uricase ranged from 9.0 to 10.5. At pH 9.2, the Km (Michaelis-Menten constant) was 204+/ 14 micromol/l (n=8) and the Ki (inhibition constant) for xanthine was 41+/-7 micromol/l (n=5). By analysing the data monitored within 5 min at 0.03 unit/ml uricase, this kinetic uricase method gave linear response to uric acid in reaction solution from 1.3 to 60 micromol/l. Aside from other common errors, 30 micromol/l xanthine in the reaction solution caused no error in this kinetic uricase method, while it caused negative error in the indirect equilibrium method by peroxidase-coupled assay of H2O2. Uric acid in clinical sera by this kinetic uricase method (Ck) closely and positively correlated with that from the indirect equilibrium method (Ce) (Ck = 0.008+1.081 x Ce, r>0.990, n=99). However, Bland Altman analysis suggested inconsistency between Ck and Ce. These results indicated that this kinetic uricase method using this uricase was reliable for serum uric acid assay with enhanced resistance to xanthine besides other common errors. PMID- 16689680 TI - Current status of pancreatic islet transplantation. AB - DM (diabetes mellitus) is a metabolic disorder of either absolute or relative insulin deficiency. Optimized insulin injections remain the mainstay life sustaining therapy for patients with T1DM (Type I DM) in 2006; however, a small subset of patients with T1DM (approx. 10%) are exquisitely sensitive to insulin and lack counter-regulatory measures, putting them at higher risk of neuroglycopenia. One alternative strategy to injected insulin therapy is pancreatic islet transplantation. Islet transplantation came of age when Paul E. Lacy successfully reversed chemical diabetes in rodent models in 1972. In a landmark study published in 2000, Shapiro et al. [A. M. Shapiro, J. R. Lakey, E. A. Ryan, G. S. Korbutt, E. Toth, G. L. Warnock, N. M. Kneteman and R. V. Rajotte (2000) N. Engl. J. Med. 343, 230-238] reported seven consecutive patients treated with islet transplants under the Edmonton protocol, all of whom maintained insulin independence out to 1 year. Substantial progress has occurred in aspects of pancreas procurement, transportation (using the oxygenated two-layer method) and in islet isolation (with controlled enzymatic perfusion and subsequent digestion in the Ricordi chamber). Clinical protocols to optimize islet survival and function post-transplantation improved dramatically with the introduction of the Edmonton protocol, but it is clear that this approach still has potential limitations. Newer pharmacotherapies and interventions designed to promote islet survival, prevent apoptosis, to promote islet growth and to protect islets in the long run from immunological injury are rapidly approaching clinical trials, and it seems likely that clinical outcomes of islet transplantation will continue to improve at the current exponential pace. PMID- 16689681 TI - Lessons on autoimmune diabetes from animal models. AB - T1DM (Type I diabetes mellitus) results from selective destruction of the insulin producing beta-cells of the pancreas by the immune system, and is characterized by hyperglycaemia and vascular complications arising from suboptimal control of blood glucose levels. The discovery of animal models of T1DM in the late 1970s and early 1980s, particularly the NOD (non-obese diabetic) mouse and the BB (BioBreeding) diabetes-prone rat, had a fundamental impact on our ability to understand the genetics, aetiology and pathogenesis of this disease. NOD and BB diabetes-prone rats spontaneously develop a form of diabetes that closely resembles the human counterpart. Early studies of these animals quickly led to the realization that T1DM is caused by autoreactive T-lymphocytes and revealed that the development of T1DM is controlled by numerous polymorphic genetic elements that are scattered throughout the genome. The development of transgenic and gene-targeting technologies during the 1980s allowed the generation of models of T1DM of reduced genetic and pathogenic complexity, and a more detailed understanding of the immunogenetics of T1DM. In this review, we summarize the contribution of studies in animal models of T1DM to our current understanding of four fundamental aspects of T1DM: (i) the nature of genetic elements affording T1DM susceptibility or resistance; (ii) the mechanisms underlying the development and recruitment of pathogenic autoreactive T-cells; (iii) the identity of islet antigens that contribute to the initiation and/or progression of islet inflammation and beta-cell destruction; and (iv) the design of avenues for therapeutic intervention that are rooted in the knowledge gained from studies of animal models. Development of new animal models will ensure continued progress in these four areas. PMID- 16689682 TI - A novel Delta9 acyl-lipid desaturase, DesC2, from cyanobacteria acts on fatty acids esterified to the sn-2 position of glycerolipids. AB - Acyl-lipid desaturases are enzymes that convert a C-C single bond into a C=C double bond in fatty acids that are esterified to membrane-bound glycerolipids. Four types of acyl-lipid desaturase, namely DesA, DesB, DesC, and DesD, acting at the Delta12, Delta15, Delta9, and Delta6 positions of fatty acids respectively, have been characterized in cyanobacteria. These enzymes are specific for fatty acids bound to the sn-1 position of glycerolipids. In the present study, we have cloned two putative genes for a Delta9 desaturase, designated desC1 and desC2, from Nostoc species. The desC1 gene is highly similar to the desC gene that encodes a Delta9 desaturase that acts on C18 fatty acids at the sn-1 position. Homologues of desC2 are found in genomes of cyanobacterial species in which Delta9-desaturated fatty acids are esterified to the sn-2 position. Heterologous expression of the desC2 gene in Synechocystis sp. PCC 6803, in which a saturated fatty acid is found at the sn-2 position, revealed that DesC2 could desaturate this fatty acid at the sn-2 position. These results suggest that the desC2 gene is a novel gene for a Delta9 acyl-lipid desaturase that acts on fatty acids esterified to the sn-2 position of glycerolipids. PMID- 16689683 TI - Scanning peptide array analyses identify overlapping binding sites for the signalling scaffold proteins, beta-arrestin and RACK1, in cAMP-specific phosphodiesterase PDE4D5. AB - The cAMP-specific phosphodiesterase PDE4D5 can interact with the signalling scaffold proteins RACK (receptors for activated C-kinase) 1 and beta-arrestin. Two-hybrid and co-immunoprecipitation analyses showed that RACK1 and beta arrestin interact with PDE4D5 in a mutually exclusive manner. Overlay studies with PDE4D5 scanning peptide array libraries showed that RACK1 and beta-arrestin interact at overlapping sites within the unique N-terminal region of PDE4D5 and at distinct sites within the conserved PDE4 catalytic domain. Screening scanning alanine substitution peptide arrays, coupled with mutagenesis and truncation studies, allowed definition of RACK1 and beta-arrestin interaction sites. Modelled on the PDE4D catalytic domain, these form distinct well-defined surface exposed patches on helices-15-16, for RACK1, and helix-17 for beta-arrestin. siRNA (small interfering RNA)-mediated knockdown of RACK1 in HEK-293 (human embryonic kidney) B2 cells increased beta-arrestin-scaffolded PDE4D5 approx. 5 fold, increased PDE4D5 recruited to the beta2AR (beta2-adrenergic receptor) upon isoproterenol challenge approx. 4-fold and severely attenuated (approx. 4-5 fold) both isoproterenol-stimulated PKA (protein kinase A) phosphorylation of the beta2AR and activation of ERK (extracellular-signal-regulated kinase). The ability of a catalytically inactive form of PDE4D5 to exert a dominant negative effect in amplifying isoproterenol-stimulated ERK activation was ablated by a mutation that blocked the interaction of PDE4D5 with beta-arrestin. In the present study, we show that the signalling scaffold proteins RACK1 and beta arrestin compete to sequester distinct 'pools' of PDE4D5. In this fashion, alterations in the level of RACK1 expression may act to modulate signal transduction mediated by the beta2AR. PMID- 16689684 TI - Novel genes encoding six kinds of three-finger toxins in Ophiophagus hannah (king cobra) and function characterization of two recombinant long-chain neurotoxins. AB - Three-finger toxins are a family of low-molecular-mass toxins (<10 kDa) having very similar three-dimensional structures. In the present study, 19 novel cDNAs coding three-finger toxins were cloned from the venom gland of Ophiophagus hannah (king cobra). Alignment analysis showed that the putative peptides could be divided into six kinds of three-finger toxins: LNTXs (long-chain neurotoxins), short-chain neurotoxins, cardiotoxins (CTXs), weak neurotoxins, muscarinic toxins and a toxin with a free SH group. Furthermore, a phylogenetic tree was established on the basis of the toxin cDNAs and the previously reported similar nucleotide sequences from the same source venom. It indicated that three-finger toxin genes in O. hannah diverged early in the course of evolution by long- and short-type pathways. Two LNTXs, namely rLNTX1 (recombinant LNTX1) and rLNTX3, were expressed and showed cytolytic activity in addition to their neurotoxic function. By comparing the functional residues, we offer some possible explanations for the differences in their neurotoxic function. Moreover, a plausible elucidation of the additonal cytolytic activity was achieved by hydropathy-profile analysis. This, to our knowledge, is the first observation that recombinant long chain alpha-neurotoxins have a CTX-like cytolytic activity. PMID- 16689685 TI - Science and scientificity. PMID- 16689686 TI - Classifying genomic sequences by sequence feature analysis. AB - Traditional sequence analysis depends on sequence alignment. In this study, we analyzed various functional regions of the human genome based on sequence features, including word frequency, dinucleotide relative abundance, and base base correlation. We analyzed the human chromosome 22 and classified the upstream, exon, intron, downstream, and intergenic regions by principal component analysis and discriminant analysis of these features. The results show that we could classify the functional regions of genome based on sequence feature and discriminant analysis. PMID- 16689687 TI - LZ complexity distance of DNA sequences and its application in phylogenetic tree reconstruction. AB - DNA sequences can be treated as finite-length symbol strings over a four-letter alphabet (A, C, T, G). As a universal and computable complexity measure, LZ complexity is valid to describe the complexity of DNA sequences. In this study, a concept of conditional LZ complexity between two sequences is proposed according to the principle of LZ complexity measure. An LZ complexity distance metric between two nonnull sequences is defined by utilizing conditional LZ complexity. Based on LZ complexity distance, a phylogenetic tree of 26 species of placental mammals (Eutheria) with three outgroup species was reconstructed from their complete mitochondrial genomes. On the debate that which two of the three main groups of placental mammals, namely Primates, Ferungulates, and Rodents, are more closely related, the phylogenetic tree reconstructed based on LZ complexity distance supports the suggestion that Primates and Ferungulates are more closely related. PMID- 16689688 TI - Characterizing the microenvironment surrounding phosphorylated protein sites. AB - Protein phosphorylation plays an important role in various cellular processes. Due to its high complexity, the mechanism needs to be further studied. In the last few years, many methods have been contributed to this field, but almost all of them investigated the mechanism based on protein sequences around protein sites. In this study, we implement an exploration by characterizing the microenvironment surrounding phosphorylated protein sites with a modified shell model, and obtain some significant properties by the rank-sum test, such as the lack of some classes of residues, atoms, and secondary structures. Furthermore, we find that the depletion of some properties affects protein phosphorylation remarkably. Our results suggest that it is a meaningful direction to explore the mechanism of protein phosphorylation from microenvironment and we expect further findings along with the increasing size of phosphorylation and protein structure data. PMID- 16689689 TI - A contact energy function considering residue hydrophobic environment and its application in protein fold recognition. AB - The three-dimensional (3D) structure prediction of proteins is an important task in bioinformatics. Finding energy functions that can better represent residue residue and residue-solvent interactions is a crucial way to improve the prediction accuracy. The widely used contact energy functions mostly only consider the contact frequency between different types of residues; however, we find that the contact frequency also relates to the residue hydrophobic environment. Accordingly, we present an improved contact energy function to integrate the two factors, which can reflect the influence of hydrophobic interaction on the stabilization of protein 3D structure more effectively. Furthermore, a fold recognition (threading) approach based on this energy function is developed. The testing results obtained with 20 randomly selected proteins demonstrate that, compared with common contact energy functions, the proposed energy function can improve the accuracy of the fold template prediction from 20% to 50%, and can also improve the accuracy of the sequence-template alignment from 35% to 65%. PMID- 16689690 TI - A branch and bound algorithm for the protein folding problem in the HP lattice model. AB - A branch and bound algorithm is proposed for the two-dimensional protein folding problem in the HP lattice model. In this algorithm, the benefit of each possible location of hydrophobic monomers is evaluated and only promising nodes are kept for further branching at each level. The proposed algorithm is compared with other well-known methods for 10 benchmark sequences with lengths ranging from 20 to 100 monomers. The results indicate that our method is a very efficient and promising tool for the protein folding problem. PMID- 16689691 TI - Preprocessing of tandem mass spectrometric data based on decision tree classification. AB - In this study, we present a preprocessing method for quadrupole time-of-flight (Q TOF) tandem mass spectra to increase the accuracy of database searching for peptide (protein) identification. Based on the natural isotopic information inherent in tandem mass spectra, we construct a decision tree after feature selection to classify the noise and ion peaks in tandem spectra. Furthermore, we recognize overlapping peaks to find the monoisotopic masses of ions for the following identification process. The experimental results show that this preprocessing method increases the search speed and the reliability of peptide identification. PMID- 16689692 TI - Constructing support vector machine ensembles for cancer classification based on proteomic profiling. AB - In this study, we present a constructive algorithm for training cooperative support vector machine ensembles (CSVMEs). CSVME combines ensemble architecture design with cooperative training for individual SVMs in ensembles. Unlike most previous studies on training ensembles, CSVME puts emphasis on both accuracy and collaboration among individual SVMs in an ensemble. A group of SVMs selected on the basis of recursive classifier elimination is used in CSVME, and the number of the individual SVMs selected to construct CSVME is determined by 10-fold cross validation. This kind of SVME has been tested on two ovarian cancer datasets previously obtained by proteomic mass spectrometry. By combining several individual SVMs, the proposed method achieves better performance than the SVME of all base SVMs. PMID- 16689693 TI - Prediction and classification of human G-protein coupled receptors based on support vector machines. AB - A computational system for the prediction and classification of human G-protein coupled receptors (GPCRs) has been developed based on the support vector machine (SVM) method and protein sequence information. The feature vectors used to develop the SVM prediction models consist of statistically significant features selected from single amino acid, dipeptide, and tripeptide compositions of protein sequences. Furthermore, the length distribution difference between GPCRs and non-GPCRs has also been exploited to improve the prediction performance. The testing results with annotated human protein sequences demonstrate that this system can get good performance for both prediction and classification of human GPCRs. PMID- 16689694 TI - Predicting the coupling specificity of GPCRs to G-proteins by support vector machines. AB - G-protein coupled receptors (GPCRs) represent one of the most important classes of drug targets for pharmaceutical industry and play important roles in cellular signal transduction. Predicting the coupling specificity of GPCRs to G-proteins is vital for further understanding the mechanism of signal transduction and the function of the receptors within a cell, which can provide new clues for pharmaceutical research and development. In this study, the features of amino acid compositions and physiochemical properties of the full-length GPCR sequences have been analyzed and extracted. Based on these features, classifiers have been developed to predict the coupling specificity of GPCRs to G-proteins using support vector machines. The testing results show that this method could obtain better prediction accuracy. PMID- 16689695 TI - Identifying G-protein coupled receptors using weighted Levenshtein distance and the nearest neighbor method. AB - G-protein coupled receptors (GPCRs) are a class of seven-helix transmembrane proteins that have been used in bioinformatics as the targets to facilitate drug discovery for human diseases. Although thousands of GPCR sequences have been collected, the ligand specificity of many GPCRs are still unknown and only one crystal structure of the rhodopsin-like family has been solved. Therefore, identifying GPCR types only from sequence data has become an important research issue. In this study, a novel technique for identifying GPCR types based on the weighted Levenshtein distance between two receptor sequences and the nearest neighbor method (NNM) is introduced, which can deal with receptor sequences with different lengths directly. In our experiments for classifying four classes (acetylcholine, adrenoceptor, dopamine, and serotonin) of the rhodopsin-like family of GPCRs, the error rates from the leave-one-out procedure and the leave half-out procedure are 0.62% and 1.24%, respectively. These results are prior to those of the covariant discriminant algorithm, the support vector machine method, and the NNM with Euclidean distance. PMID- 16689696 TI - Sequence similarity and functional relationship among eukaryotic ZIP and CDF transporters. AB - ZIP (ZRT/IRT-like Protein) and CDF (Cation Diffusion Facilitator) are two large metal transporter families mainly transporting zinc into and out of the cytosol. Several ZIP and CDF transporters have been characterized in mammals and various model organisms, such as yeast, nematode, fruit fly, and zebrafish, and many candidate genes have been identified by genome projects. Unexpected functions of ZIP and CDF transporters have been recently reported in some model organisms, leading to major advances in our understanding of the functions of mammalian counterparts. Here, we review the recent information on the sequence similarity and functional relationship among eukaryotic ZIP and CDF transporters obtained from the representative model organisms. PMID- 16689697 TI - Changes of nuclear matrix proteins following the differentiation of human osteosarcoma MG-63 cells. AB - Human osteosarcoma MG-63 cells were induced into differentiation by 5 mmol/L hexamethylene bisacetamide (HMBA). Their nuclear matrix proteins (NMPs) were selectively extracted and subjected to two-dimensional gel electrophoresis analysis. The results of protein patterns were analyzed by Melanie software. The spots of differentially expressed NMPs were excised and subjected to in situ digestion with trypsin. The maps of peptide mass fingerprinting were obtained by MALDI-TOF-MS analysis, and were submitted for NCBI database searches by Mascot tool. There were twelve spots changed remarkably during the differentiation induced by HMBA, nine of which were identified. The roles of the regulated proteins during the MG-63 differentiation were analyzed. This study suggests that the induced differentiation of cancer cells is accompanied by the changes of NMPs, and confirms the presence of some specific NMPs related to the cancer cell proliferation and differentiation. The changed NMPs are potential markers for cancer diagnosis or targets for cancer therapy. PMID- 16689698 TI - Effect of 5-azacytidine on the protein expression of porcine bone marrow mesenchymal stem cells in vitro. AB - Bone marrow-derived mesenchymal stem cells (MSCs) are pluripotent stem cells that show a vital potential in the clinical application for cell transplantation. In the present paper, proteomic techniques were used to approach the protein profiles associated with porcine bone marrow MSCs and investigate the regulation of MSC proteins on the effect of 5-azacytidine (5-aza). Over 1,700 protein species were separated from MSCs according to gel analysis. Compared with the expression profiling of control MSCs, there were 11 protein spots up-regulated and 26 down-regulated in the protein pattern of 5-aza-treated cells. A total of 21 proteins were successfully identified by MALDI-TOF-MS analysis, among which some interesting proteins, such as alpha B-crystallin, annexin A2, and stathmin 1, had been reported to involve in cell proliferation and differentiation through different signaling pathways. Our data should be useful for the future study of MSC differentiation and apoptosis. PMID- 16689699 TI - High-throughput three-dimensional gel electrophoresis for versatile utilities: a stacked slice-gel system for separation and reactions (4SR). AB - A novel high-throughput system, called the stacked slice-gel system for separation and reactions (4SR), was developed for the analysis of DNA/RNA and protein/peptide. The system provides a novel three-dimensional gel electrophoresis approach that exploits the property of stacked slice gels. It allows multiple samples simultaneously to react as well as to be separated, offering a two-dimensional (mxn) sample loading system. For this purpose, high throughput multi-micro vessels (MMVs) containing variable numbers of wells (100 wells in this paper) have been used, which are made of 25 mm square-size polyacrylamide gels. Furthermore, after electrophoretic separation, a slice gel containing a desired sample can be easily removed and proceeded to the next step. Different biological reactions as well as successive separation of products were effectively carried out dealing with DNA/RNA and protein/peptide. It shows that this system has a diversity of potentials to be developed. PMID- 16689700 TI - Predicting the nuclear localization signals of 107 types of HPV L1 proteins by bioinformatic analysis. AB - In this study, 107 types of human papillomavirus (HPV) L1 protein sequences were obtained from available databases, and the nuclear localization signals (NLSs) of these HPV L1 proteins were analyzed and predicted by bioinformatic analysis. Out of the 107 types, the NLSs of 39 types were predicted by PredictNLS software (35 types of bipartite NLSs and 4 types of monopartite NLSs). The NLSs of the remaining HPV types were predicted according to the characteristics and the homology of the already predicted NLSs as well as the general rule of NLSs. According to the result, the NLSs of 107 types of HPV L1 proteins were classified into 15 categories. The different types of HPV L1 proteins in the same NLS category could share the similar or the same nucleocytoplasmic transport pathway. They might be used as the same target to prevent and treat different types of HPV infection. The results also showed that bioinformatic technology could be used to analyze and predict NLSs of proteins. PMID- 16689701 TI - VICMpred: an SVM-based method for the prediction of functional proteins of Gram negative bacteria using amino acid patterns and composition. AB - In this study, an attempt has been made to predict the major functions of gram negative bacterial proteins from their amino acid sequences. The dataset used for training and testing consists of 670 non-redundant gram-negative bacterial proteins (255 of cellular process, 60 of information molecules, 285 of metabolism, and 70 of virulence factors). First we developed an SVM-based method using amino acid and dipeptide composition and achieved the overall accuracy of 52.39% and 47.01%, respectively. We introduced a new concept for the classification of proteins based on tetrapeptides, in which we identified the unique tetrapeptides significantly found in a class of proteins. These tetrapeptides were used as the input feature for predicting the function of a protein and achieved the overall accuracy of 68.66%. We also developed a hybrid method in which the tetrapeptide information was used with amino acid composition and achieved the overall accuracy of 70.75%. A five-fold cross validation was used to evaluate the performance of these methods. The web server VICMpred has been developed for predicting the function of gram-negative bacterial proteins (http://www.imtech.res.in/raghava/vicmpred/). PMID- 16689702 TI - PredSL: a tool for the N-terminal sequence-based prediction of protein subcellular localization. AB - The ability to predict the subcellular localization of a protein from its sequence is of great importance, as it provides information about the protein's function. We present a computational tool, PredSL, which utilizes neural networks, Markov chains, profile hidden Markov models, and scoring matrices for the prediction of the subcellular localization of proteins in eukaryotic cells from the N-terminal amino acid sequence. It aims to classify proteins into five groups: chloroplast, thylakoid, mitochondrion, secretory pathway, and "other". When tested in a five-fold cross-validation procedure, PredSL demonstrates 86.7% and 87.1% overall accuracy for the plant and non-plant datasets, respectively. Compared with TargetP, which is the most widely used method to date, and LumenP, the results of PredSL are comparable in most cases. When tested on the experimentally verified proteins of the Saccharomyces cerevisiae genome, PredSL performs comparably if not better than any available algorithm for the same task. Furthermore, PredSL is the only method capable for the prediction of these subcellular localizations that is available as a stand-alone application through the URL:http://bioinformatics.biol.uoa.gr/PredSL/. PMID- 16689703 TI - Fragrep: an efficient search tool for fragmented patterns in genomic sequences. AB - Many classes of non-coding RNAs (ncRNAs; including Y RNAs, vault RNAs, RNase P RNAs, and MRP RNAs, as well as a novel class recently discovered in Dictyostelium discoideum) can be characterized by a pattern of short but well-conserved sequence elements that are separated by poorly conserved regions of sometimes highly variable lengths. Local alignment algorithms such as BLAST are therefore ill-suited for the discovery of new homologs of such ncRNAs in genomic sequences. The Fragrep tool instead implements an efficient algorithm for detecting the pattern fragments that occur in a given order. For each pattern fragment, the mismatch tolerance and bounds on the length of the intervening sequences can be specified separately. Furthermore, matches can be ranked by a statistically well motivated scoring scheme. PMID- 16689704 TI - A quasi-physical algorithm for the structure optimization in an off-lattice protein model. AB - In this paper, we study an off-lattice protein AB model with two species of monomers, hydrophobic and hydrophilic, and present a heuristic quasi-physical algorithm. First, by elaborately simulating the movement of the smooth solids in the physical world, we find low-energy conformations for a given monomer chain. A subsequent off-trap strategy is then proposed to trigger a jump for a stuck situation in order to get out of the local minima. The algorithm has been tested in the three-dimensional AB model for all sequences with lengths of 13-55 monomers. In several cases, we renew the putative ground state energy values. The numerical results show that the proposed methods are very promising for finding the ground states of proteins. PMID- 16689706 TI - Where's the value in health care? PMID- 16689707 TI - Substantial returns to health-care spending: but do we spend too little or too much? PMID- 16689708 TI - The return on investment in health care: from 1980 to 2000. AB - OBJECTIVE: To estimate the return on US investment (ROI) in overall health as well as four specific conditions. METHODS: The study utilized three distinct approaches to "triangulate" the evidence as related to ROI in health care: 1) an estimation of the average ROI in additional health-care service expenditures in the United States for the year 2000 compared with the year 1980, based on US summaries of health expenditures and health outcomes; 2) an estimate of the ROI in Medicare services for the period from 1985 to 2000 for treatment of heart attack, stroke, type 2 diabetes, and breast cancer, based on National Long-term Care Survey data and Medicare claims; and 3) an estimate of the ROI for selected major treatment innovations for the same four conditions during the period from 1975 to 2000. RESULTS: We calculated that each additional dollar spent on overall health-care services produced health gains valued at Dollars 1.55 to Dollars 1.94 under our base case assumptions. The return on health gains associated with treatment for heart attack, stroke, type 2 diabetes, and breast cancer were Dollars 1.10, Dollars 1.49, Dollars 1.55, and Dollars 4.80, respectively, for every additional dollar spent by Medicare. The ROI for specific treatment innovations ranged from both savings in treatment costs and gains in health to gains in health valued at Dollars 1.12 to Dollars 38.00 for every additional dollar spent. CONCLUSION: The value of improved health in the US population in 2000 compared with 1980 significantly outweighs the additional health-care expenditures in 2000 compared with 1980. PMID- 16689709 TI - Economic evaluation in health care: the point of view of informed physicians. AB - OBJECTIVES: We investigated health professionals with a solid background in health-care management and economics to get their opinion and attitude on the use of economic evaluation at the policy, organizational, and professional levels of decision-making. METHODS: A 12-item questionnaire was sent to 374 Italian health care professionals who received training in economic evaluation of health-care programs in the last 10 years at the Bocconi School of Management, Milan, Italy. RESULTS: The response rate was 46.8% (175 questionnaires). All respondents stated that the basics of economic evaluation analysis must be part of the overall knowledge of health-care professionals. The usefulness of economic evaluation for professional activities was rated 3.83 (scale 1-5). Respondents stated that economic evaluation is used more for managerial decisions than for clinical ones (mean 2.89 vs. 2.74, P = 0.09). "Decisions are taken according to a short-term perspective" was the most frequently reported barrier for the actual use of economic evaluation studies, particularly by managers (76.7%). "More training in health economics" was indicated as the incentive to expand its use by the majority of both clinicians and managers (64.6%). Significantly more managers than clinicians (74.4% vs. 54.1%, P = 0.005) considered that the maximum benefits of economic evaluation are reaped at organizational level. CONCLUSIONS: Informed Italian health professionals have a positive attitude toward the principles and the techniques of economic evaluation. They show appreciation of their potential role and report making some use of them in actual decision making. PMID- 16689710 TI - Bringing health economic modeling to the 21st century. PMID- 16689711 TI - The future of health economic modeling: have we gone too far or not far enough? PMID- 16689712 TI - What decision-makers want and what they have been getting. PMID- 16689713 TI - Health resource utilization and medical care cost of acute care elderly unit patients. AB - OBJECTIVE: In this study we compared the readmissions, medical care cost, and health resource utilization (HRU) of acute care elderly (ACE) unit patients and usual medical care patients. METHODS: Retrospective case-control design was used. Patients admitted to ACE unit (n = 680) between 1999 and 2002 with primary admitting diagnosis of pneumonia, congestive heart failure, or urinary tract infection were randomly selected from the health-care system's administrative database. Equal number controls (n = 680) were selected from usual medical care services and were matched by DRG, age, ethnicity, and Charlson comorbidity score. Data on HRU, annual number of admissions before and after index admission, length of stay (LOS), and medical care cost were obtained. Bootstrap, t-test, and Wilcoxon test were used to compare cost, LOS, and number of readmissions between ACE and non-ACE unit. Multivariate log-linear and Poisson regressions were used to assess the impact of ACE unit on incremental cost and number of readmissions, respectively. RESULTS: Mean LOS was 1 day shorter for ACE unit (4.9 vs. 5.9 P = 0.01). Mean cost of ACE unit was 9.7% lower than that of non-ACE unit (Dollars 13,586 vs. Dollars 15,040, P = 0.012). Both groups had similar costs of pharmacy, diagnostic and therapeutic procedures. Multiple log-linear and Poisson regression models indicated that ACE unit patients had 21% lower cost and 11% lower annual readmissions. CONCLUSIONS: Our results confirm the hypotheses that ACE unit patients have lower medical care cost, shorter LOS, and fewer readmissions. Thus, ACE unit may be a beneficial model for improved inpatient care of elderly. PMID- 16689714 TI - Cost of hypoglycemia in patients with Type 2 diabetes in Sweden. AB - OBJECTIVES: Hypoglycemia is a common side effect of antidiabetic therapy. In addition to reducing well-being, hypoglycemic events may lead to substantial costs of medical care and lost productivity. The cost of hypoglycemia is, however, not well identified, particularly in patients with Type 2 diabetes. The purpose of this study was to assess the cost of hypoglycemia in Type 2 diabetes in Sweden. METHODS: A cost-of-illness approach, based on an incidence methodology, was used to estimate the cost of hypoglycemia in patients with Type 2 diabetes. A hypoglycemic event was defined as an episode with symptoms of low blood glucose levels during which the patient required assistance from another person. The events were divided into mild, moderate, and severe, and the incidence and costs of the different events were estimated based on data in the literature. RESULTS: Assuming that there are 300,000 patients with Type 2 diabetes in Sweden, it was estimated that 26,942 hypoglycemic events would occur annually in these patients, corresponding to a rate of 0.09 events per patient year. The total cost of hypoglycemia was, in base case, estimated at about Euro 4,250,000 (Euro 14 per patient with Type 2 diabetes) per year. Moderate hypoglycemia contributed the largest proportion of these costs. CONCLUSIONS: The results indicate that hypoglycemic events lead to substantial costs, but data are scarce and more studies are needed to better understand the cost and consequences of hypoglycemia. PMID- 16689715 TI - Linguistic validation of the US Spanish work productivity and activity impairment questionnaire, general health version. AB - INTRODUCTION: There are no measures of health-related absenteeism and presenteeism validated for use in the large and increasing US Spanish-speaking population. Before using a Spanish translation of an available English-language questionnaire, the linguistic validity of the Spanish version must be established to ensure its conceptual equivalence to the original and its cultural appropriateness. OBJECTIVE: The objective of this study was to evaluate the linguistic validity of the US Spanish version of the Work Productivity and Activity Impairment questionnaire, General Health Version (WPAI:GH). METHODS: A US Spanish translation of the US English WPAI:GH was created through a reiterative process of creating harmonized forward and back translations by independent translators. Spanish-speaking and English-speaking subjects residing in the US self-administered the WPAI:GH in their primary language and were subsequently debriefed by a bilingual (Spanish-English) interviewer. RESULTS: US Spanish subjects (N = 31) and English subjects (N = 35), stratified equally by educational level, with and without a high school degree participated in the study. The WPAI-GH item comprehension rate was 98.6% for Spanish and 99.6% for English. Response revision rates during debriefing were 1.6% for Spanish and 0.5% for English. Responses to hypothetical scenarios indicated that both language versions adequately differentiate sick time taken for health and non-health reasons and between absenteeism and presenteeism. CONCLUSION: Linguistic validity of the US Spanish translation of the WPAI:GH was established among a diverse US Spanish-speaking population, including those with minimal education. PMID- 16689716 TI - Genomes to aid in bioremediation of dry cleaning solvents, mothballs and more. PMID- 16689717 TI - Microbial community dynamics in a humic lake: differential persistence of common freshwater phylotypes. AB - In an effort to better understand the factors contributing to patterns in freshwater bacterioplankton community composition and diversity, we coupled automated ribosomal intergenic spacer analysis (ARISA) to analysis of 16S ribosomal RNA (rRNA) gene sequences to follow the persistence patterns of 46 individual phylotypes over 3 years in Crystal Bog Lake. Additionally, we sought to identify linkages between the observed phylotype variations and known chemical and biological drivers. Sequencing of 16S rRNA genes obtained from the water column indicated the presence of phylotypes associated with the Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria, TM7 and Verrucomicrobia phyla, as well as phylotypes with unknown affiliation. Employment of the 16S rRNA gene/ARISA method revealed that specific phylotypes varied independently of the entire bacterial community dynamics. Actinobacteria, which were present on greater than 95% of sampling dates, did not share the large temporal variability of the other identified phyla. Examination of phylotype relative abundance patterns (inferred using ARISA fragment relative fluorescence) revealed a strong correlation between the dominant phytoplankton succession and the relative abundance patterns of the majority of individual phylotypes. Further analysis revealed covariation among unique phylotypes, which formed several distinct bacterial assemblages correlated with particular phytoplankton communities. These data indicate the existence of unique persistence patterns for different common freshwater phylotypes, which may be linked to the presence of dominant phytoplankton species. PMID- 16689718 TI - Assessment of arbuscular mycorrhizal fungal diversity in roots of Solidago gigantea growing in a polluted soil in Northern Italy. AB - The arbuscular mycorrhizal (AM) status of Solidago gigantea was investigated in a contaminated site of Northern Italy, where the chemical industry ACNA (Associated National Chemical Companies) was active till 1999. To counteract the devastating effects of chemicals and to allow re-vegetation, soil from an uncontaminated area was used to cover the highly polluted hills of the industrial site about 25 years ago. On the basis of the current floristic features, the hill was divided into four areas. Heavy metal content in soil and in plant shoots and roots was determined by chemical analysis. The AM fungal community colonizing S. gigantea was investigated from a morphological and a molecular point of view. All plants were modestly colonized, but the fungal structures within the roots were normal. By PCR-RFLP and sequencing of 18S rDNA, 14 AM fungal types were identified: three of them were present in all the considered areas and nine appeared to be specific to certain areas. Glomus was the predominant AM genus. Our analysis demonstrates the presence and the relatively high level of AM species variety and shows how a remediation programme based on cover-soil has been efficient to restore a community of AM fungi, tolerant enough to proliferate in a still contaminated soil. PMID- 16689719 TI - Effects of water regime on archaeal community composition in Arctic soils. AB - Effects of water regime on archaeal communities in Arctic soils from Spitsbergen were studied using denaturing gradient gel electrophoresis (DGGE) of amplified 16S rRNA genes, with subsequent sequencing of amplicons and ordination analysis of binary DGGE data. Samples with major differences in soil water regime showed significant differences in their archaeal community profiles. Methanomicrobiales, Methanobacteriaceae and Methanosaeta were detectable only in environments that were wet during most of the growth season, while a novel euryarchaeotal cluster was detected only in less reduced solifluction material. Group 1.3b of Crenarchaeota had a high relative abundance within the archaeal community in a wide range of wet soils. Along a natural soil moisture gradient, changes in archaeal community composition were observed only in upper soil layers. The results indicated that members of Methanomicrobiales were relatively tolerant to soil aeration. Differences in archaeal community composition associated with soil water regime were predominant over regional and seasonal variation, and over differences between individual wetlands. The results suggest that the observed 'on-off switch' mechanism of soil hydrology for large-scale variations in methane emissions from northern wetlands is at least partly caused by differences in the community structure of organisms involved in methane production. PMID- 16689720 TI - Molecular characterization of thermostable direct haemolysin-related haemolysin (TRH)-positive Vibrio parahaemolyticus from oysters in Mangalore, India. AB - Pathogenic Vibrio parahaemolyticus strains producing either or both of a thermostable direct haemolysin (TDH) and a TDH-related haemolysin (TRH) encoded by tdh and trh genes, respectively, are isolated at a low rate from the environment. However, recently we observed that a considerable percentage of APW (alkaline peptone water) enrichment broths of oysters collected off Mangalore India, were trh(+), rather than tdh(+) by PCR. In order to further investigate the prevalence and genetic diversity of trh bearing V. parahaemolyticus in our coast, we attempted to isolate and characterize trh(+)V. parahaemolyticus from oysters. A total of 27 trh(+) strains were isolated during the period between March 2002 and February 2004, of which nine were also tdh(+). All the trh(+) isolates were positive for urease phenotype. The isolates belonged to diverse phenotypes. In order to explore the possible presence of heterogeneity in the trh gene region among trh(+)V. parahaemolyticus, a 1.5 kb region around trh gene was PCR amplified and restriction digested using selected restriction enzymes. The whole genome comparison of strains was performed by randomly amplified polymorphic DNA PCR (RAPD PCR). The PCR-RFLP results revealed fairly well conserved nature of the trh gene region studied in different serotypes. Though 11 strains were positive by PCR for a genomic fragment that has been reported to be amplified in pandemic strains, all strains were negative by group-specific PCR (GS-PCR), orf8 PCR and showed a different RAPD pattern compared with pandemic strains. The results suggest that genetically diverse V. parahaemolyticus carrying virulence genes are associated with the aquatic environment in this region. PMID- 16689721 TI - Effects of management regime and plant species on the enzyme activity and genetic structure of N-fixing, denitrifying and nitrifying bacterial communities in grassland soils. AB - Management by combined grazing and mowing events is commonly used in grasslands, which influences the activity and composition of soil bacterial communities. Whether observed effects are mediated by management-induced disturbances, or indirectly by changes in the identity of major plant species, is still unknown. To address this issue, we quantified substrate-induced respiration (SIR), and the nitrification, denitrification and free-living N(2)-fixation enzyme activities below grass tufts of three major plant species (Holcus lanatus, Arrhenatherum elatius and Dactylis glomerata) in extensively or intensively managed grasslands. The genetic structures of eubacterial, ammonia oxidizing, nitrate reducing, and free-living N(2)-fixing communities were also characterized by ribosomal intergenic spacer analysis, and denaturing gradient gel electrophoresis (DGGE) or restriction fragment length polymorphism (RFLP) targeting group-specific genes. SIR was not influenced by management and plant species, whereas denitrification enzyme activity was influenced only by plant species, and management-plant species interactions were observed for fixation and nitrification enzyme activities. Changes in nitrification enzyme activity were likely largely explained by the observed changes in ammonium concentration, whereas N availability was not a major factor explaining changes in denitrification and fixation enzyme activities. The structures of eubacterial and free-living N(2) fixing communities were essentially controlled by management, whereas the diversity of nitrate reducers and ammonia oxidizers depended on both management and plant species. For each functional group, changes in enzyme activity were not correlated or were weakly correlated to overall changes in genetic structure, but around 60% of activity variance was correlated to changes in five RFLP or DGGE bands. Although our conclusions should be tested for other ecosystems and seasons, these results show that predicting microbial changes induced by management in grasslands requires consideration of management-plant species interactions. PMID- 16689722 TI - The role of quorum sensing mediated developmental traits in the resistance of Serratia marcescens biofilms against protozoan grazing. AB - Resistance against protozoan grazers is a crucial factor that is important for the survival of many bacteria in their natural environment. However, the basis of resistance to protozoans and how resistance factors are regulated is poorly understood. In part, resistance may be due to biofilm formation, which is known to protect bacteria from environmental stress conditions. The ubiquitous organism Serratia marcescens uses quorum sensing (QS) control to regulate virulence factor expression and biofilm formation. We hypothesized that the QS system of S. marcescens also regulates mechanisms that protect biofilms against protozoan grazing. To investigate this hypothesis, we compared the interactions of wild type and QS mutant strains of S. marcescens biofilms with two protozoans having different feeding types under batch and flow conditions. Under batch conditions, S. marcescens forms microcolony biofilms, and filamentous biofilms are formed under flow conditions. The microcolony-type biofilms were protected from grazing by the suspension feeder, flagellate Bodo saltans, but were not protected from the surface feeder, Acanthamoeba polyphaga. In contrast, the filamentous biofilm provided protection against A. polyphaga. The main findings presented in this study suggest that (i) the QS system is not involved in grazing resistance of S. marcescens microcolony-type biofilms; (ii) QS in S. marcescens regulates antiprotozoan factor(s) that do not interfere with the grazing efficiency of the protozoans; and (iii) QS-controlled, biofilm-specific differentiation of filaments and cell chains in biofilms of S. marcescens provides an efficient mechanism against protozoan grazing. PMID- 16689723 TI - The Acinetobacter outer membrane protein A (OmpA) is a secreted emulsifier. AB - Acinetobacter strains use hydrophobic carbon sources and most of them are efficient oil degraders. They secrete a variety of emulsifiers which are efficient in producing and stabilizing oil-in-water emulsions. The bioemulsifier of Acinetobacter radioresistens KA53 (Alasan) is a high-mass complex of proteins and polysaccharides. The major emulsification activity of this complex is associated with a 45 kDa protein (AlnA), which is homologous to the outer membrane protein OmpA. The emulsification ability of AlnA depends on the presence of hydrophobic residues in the four loops spanning the transmembrane domains. The finding of a secreted OmpA was unexpected, in view of the fact that this protein is essential in all Gram-negative bacteria, has four trans-membrane domains and is considered to be an integral structural component of the outer membrane. However, secretion of an OmpA with emulsifying ability could be of physiological importance in the utilization of hydrophobic substrates as carbon sources. Here we examined the possibility that secretion of OmpA with emulsifying activity is a general property of the oil-degrading Acinetobacter strains. The results indicate that OmpA is secreted in five strains of Acinetobacter, including strain Acinetobacter sp. ADP1 whose genome has been sequenced. The ompA genes of ADP1 and an additional strain, Acinetobacter sp. V-26 were cloned and sequenced. Structure analysis of the sequence of the two proteins indicated the existence of the hydrophobic regions, previously shown to be responsible for the emulsification activity of AlnA. Further examination of the recombinant OmpA proteins indicated that they are, indeed, strong emulsifiers, even when produced in Escherichia coli. The finding that Acinetobacter OmpA has emulsifying activity and that it is secreted in five strains of Acinetobacter may be physiologically significant and suggests the involvement of this protein in biodegradation of hydrophobic substrates, including hydrocarbons. PMID- 16689724 TI - Analysis of fimbrial gene clusters and their expression in enterohaemorrhagic Escherichia coli O157:H7. AB - The sequence of two enterohaemorrhagic Escherichia coli (EHEC) O157:H7 strains reveals the possession of at least 16 fimbrial gene clusters, many of the chaperone/usher class. The first part of this study examined the distribution of these clusters in a selection of EHEC/EPEC (enteropathogenic E. coli) serotypes to determine if any were likely to be unique to E. coli O157:H7. Six of the clusters, as determined by the presence of amplified main subunit or usher gene sequences, were detected only in the E. coli O157 and O145 serotypes tested. With the exception of one serotype O103 strain that contained an lpf2 cluster, lpf sequences were only detected in E. coli O157 of the serotypes tested. Expression from each cluster was measured by the construction of chromosomally integrated lacZ promoter fusions and plasmid-based eGFP fusions in E. coli O157:H7. This analysis demonstrated that the majority (11/15) of main fimbrial subunit genes were not expressed under the majority of conditions tested in vitro. One of the clusters showing promoter activity, loc8, has a temperature expression optimum indicating a possible role outside the host. From the presence of pseudogenes in three of the clusters, the lack of FimH-like minor adhesins in the clusters and their limited expression in vitro, it would appear that E. coli O157:H7 has a limited repertoire of expressed functional fimbriae. This restricted selection of fimbriae may be important in bringing about the tropism E. coli O157:H7 demonstrates for the terminal rectum of cattle. PMID- 16689725 TI - Bacterial diversity is determined by volume in membrane bioreactors. AB - It has been proposed that established models and theories developed in classical ecology could be employed to greatly improve the optimization of wastewater treatment plants (WWTP) by placing the microbiological component onto a model predictive basis. In particular, this could be achieved by better understanding bacterial community assembly and development. The species-area relationship is one of the oldest biological laws and has been used to describe spatial diversity patterns in contiguous habitats and on islands. In the current study, bacterial communities in seven membrane bioreactors (MBR), of increasing size, located across the UK were sampled. A significant linear relationship between bacterial taxa richness and reactor size was observed and was similar to classical species area relationships of larger organisms colonizing oceanic islands. Rank-abundance plots revealed a gradient of greater evenness in community structure as MBR volume increased. Application of the Raup and Crick probability-based similarity index indicated a strong role for dispersal in MBR colonization and community structure. Our findings demonstrate that the MBR sampled behaved like islands with respect to bacterial colonization in accordance with the theory of island biogeography. In addition this study provides further evidence that biodiversity at the bacterial level is more similar to that of animals and plants than previously postulated. PMID- 16689726 TI - Genetic variability in environmental isolates of Legionella pneumophila from Comunidad Valenciana (Spain). AB - Legionella pneumophila is associated to recurrent outbreaks in several Comunidad Valenciana (Spain) localities, especially in Alcoi, where social and climatic conditions seem to provide an excellent environment for bacterial growth. We have analysed the nucleotide sequences of three loci from 25 environmental isolates from Alcoi and nearby locations sampled over 3 years. The analysis of these isolates has revealed a substantial level of genetic variation, with consistent patterns of variability across loci, and comparable to that found in a large, European-wide sampling of clinical isolates. Among the tree loci studied, fliC showed the highest level of nucleotide diversity. The analysis of isolates sampled in different years revealed a clear differentiation, with samples from 2001 being significantly distinct from those obtained in 2002 and 2003. Furthermore, although linkage disequilibrium measures indicate a clonal nature for population structure in this sample, the presence of some recombination events cannot be ruled out. PMID- 16689727 TI - Amplification of low quantity bacterial RNA for microarray studies: time-course analysis of Leptospirillum ferrooxidans under nitrogen-fixing conditions. AB - We have developed a method for the amplification of low quantity total bacterial RNA for DNA microarrays analysis. Current methods are based on the linear amplification by the in vitro transcription from the T7 promoter, similar to that used for eukaryotic mRNA amplification. For the incorporation of T7 promoter, the prokaryotic RNA must be enzymatically modified for the incorporation of a polyA tail at the 3' end to emulate the eukaryotic mRNA. The method we describe and validate herein avoids this step by the direct and random incorporation of the T7 promoter. From 500 ng of total bacterial RNA, we obtained 130-150 microg of antisense RNA, such products being good substrate for fluorescent labelling and DNA microarray analysis. The method was validated with bacterial samples from which it is very difficult to obtain sufficient amounts and quality of total RNA for global gene expression analysis. This is critical for low cell density growing microorganisms, environmental samples, or many extremophiles where the composition of the cultural media severely affects the RNA yield, like in the case of the acidophile and iron oxidizer Gram-negative bacterium Leptospirillum ferrooxidans. We further validated our amplification method in parallel experiments with non-amplified RNA by following the expression of the L. ferrooxidans nif regulon along the time-course of growth. PMID- 16689728 TI - Algae-bacteria interactions and their effects on aggregation and organic matter flux in the sea. AB - Aggregation of algae, mainly of diatoms, is an important process in marine pelagic systems, often terminating phytoplankton blooms and leading to the sinking of particulate organic matter in the form of marine snow. This process has been studied extensively, but the specific role of heterotrophic bacteria has largely been neglected, mainly because field studies and most experimental work were performed under non-axenic conditions. We tested the hypothesis that algae bacteria interactions are instrumental in aggregate dynamics and organic matter flux. A series of aggregation experiments has been carried out in rolling tanks with two marine diatoms typical of temperate regions (Skeletonema costatum and Thalassiosira rotula) in an axenic treatment and one inoculated with marine bacteria. Exponentially growing S. costatum and T. rotula exhibited distinctly different aggregation behavior. This was reflected by their strikingly different release of dissolved organic matter (DOM), transparent exopolymer particles (TEP) and protein-containing particles (CSP), as well as their bacterial biodegradability and recalcitrance. Cells of S. costatum aggregated only little and their bacterial colonization remained low. Dissolved organic matter, TEP and CSP released by this alga were largely consumed by free-living bacteria. In contrast, T. rotula aggregated rapidly and DOM, TEP and CSP released resisted bacterial consumption. Experiments conducted with T. rotula cultures in the stationary growth phase, however, showed rapid bacterial colonization and decomposition of algal cells. Our study highlights the importance of heterotrophic bacteria to control the development and aggregation of phytoplankton in marine systems. PMID- 16689729 TI - Contamination routes of Clostridium botulinum in the honey production environment. AB - Factors influencing Clostridium botulinum contamination in the honey production environment were evaluated in a 3-year survey. A number of 1,168 samples from 100 apiaries and related facilities were analysed for the presence of C. botulinum types A, B, E and F, using multiplex polymerase chain reaction targeted to botA, botB, botE and botF genes. Production methods and environmental factors were registered using a questionnaire and by personal observation. Clostridium botulinum was shown to be a common finding throughout the whole honey production chain, and the type most frequently detected was group I type B. In a pulsed field gel electrophoresis (PFGE) analysis of 202 group I type B isolates, only six different PFGE profiles were observed, of which two clearly distinct profiles predominated. This may indicate the existence of at least two different genetic lineages. The high prevalence of C. botulinum in soil and in samples associated with beeswax suggests the accumulation of soil-derived botulinal spores in wax. Additionally, according to Spearman's rank order correlation and multivariate analysis, production hygiene-dependent factors have a significant influence on the contamination, and thus the number and frequency of C. botulinum spores in honey could possibly be diminished by increasing hygienic level in honey production. PMID- 16689730 TI - The galactophilic lectin, LecA, contributes to biofilm development in Pseudomonas aeruginosa. AB - LecA (PA-IL) is a cytotoxic lectin and adhesin produced by Pseudomonas aeruginosa which binds hydrophobic galactosides with high specificity and affinity. By using a lecA-egfp translation fusion and immunoblot analysis of the biofilm extracellular matrix, we show that lecA is expressed in biofilm-grown cells. In static biofilm assays on both polystyrene and stainless steel, biofilm depth and surface coverage was reduced by mutation of lecA and enhanced in the LecA overproducing strain PAO-P47. Biofilm surface coverage by the parent strain, PAO P47 but not the lecA mutant on steel coupons was also inhibited by growth in the presence of either isopropyl-beta-D-thiogalactoside (IPTG) or p-nitrophenyl-alpha D-galactoside (NPG). Furthermore, mature wild-type biofilms formed in the absence of these hydrophobic galactosides could be dispersed by the addition of IPTG. In contrast, addition of p-nitrophenyl-alpha-L-fucose (NPF) which has a high affinity for the P. aeruginosa LecB (PA-IIL) lectin had no effect on biofilm formation or dispersal. Planktonic growth of P. aeruginosa PAO1 was unaffected by the presence of IPTG, NPG or NPF, nor was the strain able to utilize these sugars as carbon sources, suggesting that the observed effects on biofilm formation were due to the competitive inhibition of LecA-ligand binding. Similar results were also obtained for biofilms grown under dynamic flow conditions on steel coupons, suggesting that LecA contributes to P. aeruginosa biofilm architecture under different environmental conditions. PMID- 16689731 TI - Perkinsoide chabelardi n. gen., a protozoan parasite with an intermediate evolutionary position: possible cause of the decrease of sardine fisheries? AB - Phenotypic scrutiny on the life cycle of Icthyodinium chabelardi (Perkinsoide chabelardi n. gen.) based on ultrastructural techniques, and molecular phylogenetic analysis of RNA gene sequences, were carried out in order to elucidate the taxonomic position of this parasite. The absence of plastid, presence of trichocysts, and chromosomes or chromatin condensed and low in number, suggested that this protozoan could be considered a dinoflagellate syndinial parasite. However, the life cycle, schizogonic divisions and structure of schizonts inside the host, the nuclei without the typical dinoflagellate appearance, presence of rhoptrias-like structures, a possible pseudo-conoid, and the biflagellated spore, resembled those of the genus Perkinsus. Phylogenetic analysis of genes transcribing for the RNA forming the small subunit and the large subunit suggests that this parasite has an ambiguous evolutionary position within the group formed by dinoflagellates, perkinsids and syndinials. Because of differences with dinoflagellates and similarities with perkinsids, we propose to change the generic name to P. chabelardi n. gen. High stationary infection prevalence on Sardina pilchardus eggs was observed. This protozoan parasite caused the death of all the infected sardine eggs, and therefore a high impact in the recruitment of this fishery in the Atlantic coast is expected. PMID- 16689732 TI - Modelled and measured dynamics of viruses in Arctic winter sea-ice brines. AB - We describe a model based on diffusion theory and the temperature-dependent mechanism of brine concentration in sea ice to argue that, if viruses partition with bacteria into sea-ice brine inclusions, contact rates between the two can be higher in winter sea ice than in seawater, increasing the probability of infection and possible virus production. To examine this hypothesis, we determined viral and bacterial concentrations in select winter sea-ice horizons using epifluorescence microscopy. Viral concentrations ranged from 1.6 to 82 x 10(6) ml(-1) of brine volume of the ice, with highest values in brines from coldest (-24 to -31 degrees C) ice horizons. Calculated virus-bacteria contact rates in underlying -1 degrees C seawater were similar to those in brines of -11 degrees C ice but up to 600 times lower than those in ice brines at or below -24 degrees C. We then incubated native bacterial and viral assemblages from winter sea ice for 8 days in brine at a temperature (-12 degrees C) and salinity ( approximately 160 psu) near expected in situ values, monitoring their concentrations microscopically. While different cores yielded different results, consistent with known spatial heterogeneity in sea ice, these experiments provided unambiguous evidence for viral persistence and production, as well as for bacterial growth, in -12 degrees C brine. PMID- 16689733 TI - Identification of differentially expressed genes in Tetrahymena thermophila in response to dichlorodiphenyltrichloroethane (DDT) by suppression subtractive hybridization. AB - The insecticide dichlorodiphenyltrichloroethane (DDT) is persistent in the environment, and continues to cause health problems. Tetrahymena has potential as a model organism for assaying low levels of DDT and for analysing the mechanisms of its toxicity. We constructed the suppression subtractive hybridization library of T. thermophila exposed to DDT, and screened out 90 Expressed Sequence Tags whose expressions were significantly up- or downregulated with DDT treatment. From this, a series of important genes related to the DDT metabolism and detoxification were discovered, such as P450 gene, glutathione S-transferase gene and sterol carrier protein 2 gene. Furthermore, their expressions under different concentrations of DDT treatment were detected by real-time fluorescent quantitative PCR. The results show that Tetrahymena is a relevant and useful model organism for detecting DDT in the environment and for discovering biomarkers that can be used to develop specific bio-reporters at the molecular and genomic levels. PMID- 16689734 TI - Amoebae promote persistence of epidemic strains of MRSA. AB - The control of healthcare-associated methicillin-resistant Staphylococcus aureus (MRSA) infection is of concern worldwide. Given the evidence that several pathogenic species replicate within amoebae and emerge more virulent and more resistant and the abundance of amoebae in healthcare settings, we investigated interactions of Acanthamoeba polyphaga with epidemic MRSA isolates. MRSA proliferated in the presence of amoebae, attributable partly to intracellular replication. Following 24 h of co-culture, confocal microscopy revealed that c. 50% amoebae had viable MRSA within phago-lysosomes and 2% of amoebae were heavily infected with viable cocci throughout the cytoplasm. Infection control strategies should recognize the contribution of protozoa. PMID- 16689736 TI - Deciphering the mystery of combined factor V and factor VIII deficiency. PMID- 16689737 TI - Fibrin structure and wound healing. AB - Fibrinogen and fibrin play an important role in blood clotting, fibrinolysis, cellular and matrix interactions, inflammation, wound healing, angiogenesis, and neoplasia. The contribution of fibrin(ogen) to these processes largely depends not only on the characteristics of the fibrin(ogen) itself, but also on interactions between specific-binding sites on fibrin(ogen), pro-enzymes, clotting factors, enzyme inhibitors, and cell receptors. In this review, the molecular and cellular biology of fibrin(ogen) is reviewed in the context of cutaneous wound repair. The outcome of wound healing depends largely on the fibrin structure, such as the thickness of the fibers, the number of branch points, the porosity, and the permeability. The binding of fibrin(ogen) to hemostasis proteins and platelets as well as to several different cells such as endothelial cells, smooth muscle cells, fibroblasts, leukocytes, and keratinocytes is indispensable during the process of wound repair. High-molecular weight and low-molecular-weight fibrinogen, two naturally occurring variants of fibrin, are important determinants of angiogenesis and differ in their cell growth stimulation, clotting rate, and fibrin polymerization characteristics. Fibrin sealants have been investigated as matrices to promote wound healing. These sealants may also be an ideal delivery vehicle to deliver extra cells for the treatment of chronic wounds. PMID- 16689738 TI - Unveiling the new face of fibronectin in thrombosis and hemostasis. PMID- 16689739 TI - Characterization of fibronectin assembly by platelets adherent to adsorbed laminin-111. AB - BACKGROUND: Various types of laminin (LN) are ubiquitous components of basement membrane and exposed to blood upon localized damage of vascular endothelial cells. Fibronectin is a plasma protein that is insolubilized into fibrils in a regulated fashion by, for example, lysophosphatidic acid (LPA)-stimulated fibroblasts or platelets spread on supportive adhesive ligands. OBJECTIVE: To study assembly of plasma fibronectin by LPA-activated platelets adherent to LN 111 via alpha6beta1 integrin. RESULTS: Platelets adherent to LN-111-bound plasma fibronectin or its N-terminal 70 kD fragment in fibrillar arrays at the periphery of spread platelets under static but not shear conditions. Formation of fibronectin arrays under static conditions was inhibited by co-incubation with the N-terminal 70 kD fragment or with a 49-amino acid peptide that binds to the N terminal region of fibronectin. Approximately 7000 fibronectin dimers bound per adherent platelet with a K(d) of 50 nm. Bound 70 kD fragment was readily solubilized with deoxycholate (DOC), whereas bound fibronectin became progressively insoluble. Bound 70 kD fragment became resistant to DOC extraction after treatment with a cell-impermeable, reducible crosslinker. Crosslinked 70 kD fragment was found in a high molecular weight complex. As with fibroblasts, signaling molecules modulating actin cytoskeletal organization controlled expression of binding sites for the N-terminal 70 kD region of fibronectin on adherent platelets. CONCLUSIONS: These results indicate that platelets adherent to LN-111 via alpha(6)beta(1) support subsequent assembly of fibronectin, but possibly only under conditions of intermittent or stagnant blood flow. PMID- 16689740 TI - A new tool to further explore the role of ADAMTS-13 in health and disease. PMID- 16689741 TI - ADAMTS-13 plasma level determination uncovers antigen absence in acquired thrombotic thrombocytopenic purpura and ethnic differences. AB - BACKGROUND: The recently discovered plasma enzyme ADAMTS-13 cleaves the A2-domain of von Willebrand factor (VWF). A defective cleaving protease results in unusually large VWF multimers, which cause thrombotic thrombocytopenic purpura (TTP). AIM: Analysis of the ADAMTS-13 antigen levels in TTP patients compared with normal donors. METHODS: An antigen ELISA test was built, based on high affinity anti-ADAMTS-13 monoclonal antibodies, which were generated using genetic immunization. RESULTS: Specificity of the ADAMTS-13 antigen test was confirmed, as (i) plasma from a patient with acquired TTP but presenting without inhibitor did not contain antigen and (ii) the binding of recombinant ADAMTS-13 was inhibited by increasing amounts of normal plasma. The assay is sensitive as it can detect antigen levels as low as 1.6% of normal. The concentration in normal pooled human plasma was determined (1.03 +/- 0.15 microg mL(-1)) and arbitrarily set to 1 U mL(-1). The antigen levels in congenital TTP samples (34 +/- 21 mU mL( 1), n = 2), as well as in samples from patients with acquired TTP (231 +/- 287 mU mL(-1), n = 11), were clearly reduced when compared with normal Caucasian donors (951 +/- 206 mU mL(-1), n = 16). Remarkably, normal Chinese donors have a significantly lower antigen titer (601 +/- 129 mU mL(-1), n = 15), when compared with normal Caucasians. CONCLUSIONS: Our results show that acquired TTP patients suffer mainly from ADAMTS-13 antigen depletion, thereby indicating the importance of ADAMTS-13 antigen determination in diagnosis and patient follow-up. PMID- 16689742 TI - Urgent reversal of warfarin with prothrombin complex concentrate: where are the evidence-based data? PMID- 16689743 TI - Urgent reversal of warfarin with prothrombin complex concentrate. AB - BACKGROUND: When life-threatening bleeding occurs in patients on warfarin, timely reversal becomes imperative. In the USA, warfarin effect is commonly reversed with fresh frozen plasma (FFP). The use of FFP is complicated by delays in correction, volume overload and often, inadequate correction. OBJECTIVE: Evaluate the feasibility and efficacy of a protocol for rapid administration of prothrombin complex concentrate (PCC) in the setting of the urgent need for reversal of warfarin. METHODS/PATIENTS: We instituted a policy for rapid delivery and administration of PCC. Appropriate patients received 25-50 U kg(-1) of PCC. The prothrombin time (PT)/International Normalized Ratios (INR) was recorded before and immediately after dosing, and 24 h postdosing. Patients requiring surgical interventions were cleared for the operating room (OR) immediately. Fifty-eight patients were treated, with a median age of 75.5 years (range 26-92). RESULTS: The median INR on presentation was 3.8 (1.4-52.8). Immediately following PCC administration the median INR was 1.3 (0.9-5.7), only two patients with INRs exceeding 2.0. The benefit was maintained at 24 h with a median INR of 1.5 (1.1 3.4). Four patients experienced thrombotic events during their hospitalization, (two deep vein thrombosis, two non-q-wave myocardial infarction) although none was attributed to PPC therapy. CONCLUSIONS: PCC administration is an effective treatment modality for the correction of warfarin anticoagulation in the urgent setting. Advantages over FFP include more timely correction, absence of volume overload and potentially more complete correction. Broader use of PCC in this setting appears to be appropriate. PMID- 16689744 TI - Elusive proatherothrombotic role of Lp(a): a new direction? PMID- 16689745 TI - Lipoprotein(a) as a modifier of fibrin clot permeability and susceptibility to lysis. PMID- 16689746 TI - Circulating endothelial cells and endothelial progenitor cells after angioplasty: news from the endothelial rescue squad. PMID- 16689747 TI - Endothelial injury induced by coronary angioplasty triggers mobilization of endothelial progenitor cells in patients with stable coronary artery disease. PMID- 16689748 TI - Effects of older age on fibrin D-dimer, C-reactive protein, and other hemostatic and inflammatory variables in men aged 60-79 years. AB - BACKGROUND: Previous studies have suggested that several hemostatic and inflammatory variables, which are risk predictors for arterial or venous thrombosis, increase with age. However, there is a lack of data from large population studies for reliable estimates of reference ranges. OBJECTIVES: To establish reliable reference ranges of hemostatic and inflammatory variables for 5-year age groups in older men and their implications for pathogenesis and diagnosis. PATIENTS AND METHODS: A total of 3861 men aged 60-79 years at the 20 years follow-up of the British Regional Heart Study. RESULTS: Several variables increased with age. The greatest median increases between 60-64 and 75-79 years age groups were observed for fibrin D-dimer (91%) and C-reactive protein (CRP) (57%). Significant median increases were also observed for von Willebrand factor antigen (23%), tissue plasminogen activator antigen (11%), factor VIII (10%), and fibrinogen (8%). In contrast, levels of classical cardiovascular risk factors neither decreased nor increased substantially with age, with the exception of systolic blood pressure (median increase 10%). CONCLUSIONS: The exponential increases in risk of arterial and venous thrombotic events in men between age 60 and 79 years (when most such events occur) may be related in part to increasing activation of blood coagulation, fibrinolysis, and inflammation; possibly related to the increasing inflammatory burden of both atherosclerotic and non-vascular disease. These increases also have implications for diagnosis of suspected acute venous thromboembolism (D-dimer), and recently proposed screening for prediction of coronary heart disease risk and detection of occult disease (CRP). PMID- 16689749 TI - Renal transplant recipients are at high risk for both symptomatic and asymptomatic deep vein thrombosis. AB - BACKGROUND: Venous thromboembolism (VTE) is one of the thrombotic complications that can occur in patients receiving renal transplantation (RT). The prevalence of VTE in RT patients is, however, undefined. OBJECTIVES: To evaluate the rate of a first episode of VTE in a series of 538 consecutive RT recipients admitted to our institution, the timing of occurrence of the thromboembolic events after transplantation, and the rate of recurrence after thromboprophylaxis withdrawal. Risk factors for recurrence were also evaluated, particularly in relation to the type of the first event (symptomatic or asymptomatic). RESULTS: During follow-up, 47 of 518 patients (28 males, 19 females; 9.1%) developed a first episode of VTE at a median time of 17 months (range 1-165 months) after kidney transplantation. Cancer was associated with the occurrence of VTE (odds ratio 4.8). Seventeen of 43 patients (39.5%) with deep vein thrombosis were asymptomatic and the diagnosis was made during routine ultrasound examination. Twenty-two patients (46.8%) experienced a recurrence of VTE. A relevant rate of recurrence was documented amongst patients with a first episode of both symptomatic (53%) and asymptomatic (23.5%) VTE. CONCLUSION: This study confirms that RT patients are at high risk of symptomatic and asymptomatic VTE and that this risk persists even after several years. Patients who experience VTE are at high risk of recurrence after thromboprophylaxis withdrawal. PMID- 16689750 TI - G-CSF-mobilized peripheral blood mononuclear cells from diabetic patients augment neovascularization in ischemic limbs but with impaired capability. AB - BACKGROUND: Autologous transplantation of mobilized peripheral blood mononuclear cells (M-PBMNCs) is a novel approach to improve critical limb ischemia (CLI) in diabetes. However, endothelial progenitor cells (EPCs) from diabetes are dysfunctional and impaired in ischemia-induced neovascularization. OBJECTIVE: This study aimed to confirm the compromised efficiency of diabetic M-PBMNCs in therapeutic neovascularization, and to determine the underlying mechanisms of this impairment. METHODS: Diabetic M-PBMNCs from 17 diabetic patients or healthy controls, or phosphate-buffered saline (PBS) were injected into the ischemic limbs of streptozotocin-induced diabetic nude mice. The limb blood perfusion, ambulatory score, ischemia damage, capillary/fiber ratio, arteriole density, collateral vessel formation, and pericytes recruitment were evaluated between these three groups. Non-invasive real time image and histopathology were used to detect the in vivo role of transplanted M-PBMNCs. Proliferation and adhesion of EPCs were assayed. In vitro vascular network incorporation and matrigel plug assay were used to test the pro-neovascularization role of M-PBMNCs. RESULTS: Transplantation of diabetic M-PBMNCs also improved neovascularization, but to a lesser extent from that observed with non-diabetic ones. This was associated with the impairment of diabetic M-PBMNCs capacity to differentiate into EPCs, to incorporate into vessel-like tubules in vitro, to participate in vascular-like structure formation in a subcutaneous matrigel plug, and to stimulate the recruitment of pericytes/smooth muscle cells. In addition, there was impairment in vasculogenesis, which was related to the reduced adhesion ability of EPCs from diabetic M-PBMNCs. CONCLUSIONS: Diabetes reduced the capacity of M-PBMNCs to augment neovascularization in ischemia. PMID- 16689751 TI - Two consecutive high-fat meals affect endothelial-dependent vasodilation, oxidative stress and cellular microparticles in healthy men. AB - BACKGROUND: A large body of evidence has accumulated indicating a relation between postprandial hyperglycemia and hypertriglyceridemia, and the risk of cardiovascular disease. OBJECTIVE: We studied possible mechanisms underlying the postprandial proatherogenic state by exposing healthy males to two consecutive high-fat mixed meals. PATIENTS/METHODS: Seventeen healthy males [age 25.4 +/- 3 years, body mass index 23.6 +/- 2 kg m(-2)] were studied during two randomized visits. During the meal visit, subjects consumed standardized meals (50 g of fat, 55 g of carbohydrates and 30 g of proteins) as breakfast and 4 h later as lunch. During the control visit, subjects remained fasted. Prior to each blood collection (before and every 2 h after the first meal), flow-mediated dilation (FMD) of the brachial artery was measured. RESULTS: Although within the normal range, postprandial plasma glucose and triacylglycerol concentrations increased significantly, especially after the second meal, as compared with baseline (4.8 +/- 0.3 to 5.4 +/- 0.4, 0.8 +/- 0.2 to 1.7 +/- 0.7 mmol L(-1), respectively; both P < 0.05) and the fasting visit. After the second meal, FMD was significantly impaired (6.9% vs. 3.7%, P < 0.05) whereas oxidized low-density lipoprotein (oxLDL)/LDL cholesterol ratio and malondialdehyde concentrations were markedly elevated (both P < 0.01). Finally, an increase in total microparticle (MP) numbers was observed during the meal visit (P < 0.05). CONCLUSIONS: In healthy males, after two consecutive fat-rich meals, mild elevations in plasma glucose and triacylglycerol were paralleled by impaired FMD, increased markers of oxidative stress and circulating MPs, in particular, after the second meal. These findings may have consequences for subjects with postprandial dysmetabolism, including those with Type 2 diabetes. PMID- 16689752 TI - Disposition of tissue factor pathway inhibitor during cardiopulmonary bypass. AB - BACKGROUND: The tissue factor (TF) factor (F) VIIa complex activates coagulation FIX and FX to initiate coagulation, and also cleaves protease activated receptors (PARs) to initiate inflammatory processes in vascular cells. Tissue factor pathway inhibitor (TFPI) is the only specific inhibitor of the TF-FVIIa complex, regulating both its procoagulant and pro-inflammatory properties. Upon heparin infusion during cardiopulmonary bypass (CPB), a heparin releasable pool of endothelial associated TFPI circulates in plasma. Following protamine neutralization of heparin, the plasma TFPI level decreases, but does not return completely to baseline, suggesting that during CPB a fraction of the plasma TFPI becomes heparin-independent. We have investigated the structural and functional properties of plasma TFPI during CPB to further characterize how TFPI is altered during this procedure. METHODS: We enrolled 17 patients undergoing first-time cardiac surgery involving CPB. Plasma samples were obtained at baseline, 5 min and 1 h after start of CPB (receiving heparin), 10 min after protamine administration (off CPB) and 24 h following surgery. Samples were analyzed for full-length and free (non-lipoprotein bound) TFPI antigen by enzyme-linked immunosorbent assay (ELISA) and for TFPI anticoagulant activity using an amidolytic assay. Western blot analysis was used to identify TFPI species of varying molecular weights in three additional patients. Dunnett's test for post hoc comparisons was used for statistical analysis. RESULTS: The ELISA and Western blot data indicated that an increase in full-length TFPI accounted for most of the heparin releasable TFPI. Following heparin neutralization with protamine, the full-length TFPI antigen returned to baseline levels while the free TFPI antigen and the total plasma TFPI activity remained elevated. This was associated with the appearance of a new 38 kDa form of plasma TFPI identified by Western blot analysis. The 38 kDa form of TFPI did not react with an antibody directed against the C-terminal region of TFPI indicating it has undergone proteolysis within this region. All TFPI measurements returned to baseline 24 h following CPB. CONCLUSIONS: During CPB the full-length form of TFPI is the predominant form in plasma because of its prompt release from the endothelial surface following heparin administration. Upon heparin neutralization with protamine, full-length TFPI redistributes back to the endothelial surface. However, a new 38 kDa TFPI fragment is generated during CPB and remains circulating in plasma, indicating that TFPI undergoes proteolytic degradation during CPB. This degradation may result in a decrease in endothelium-associated TFPI immediately post-CPB, and may contribute to the procoagulant and proinflammatory state that often complicates CPB. PMID- 16689753 TI - Markers of hypercoagulability and inflammation predict mortality in patients with heart failure. AB - BACKGROUND AND AIMS: Plasma levels of inflammatory markers are increased in chronic heart failure (HF) and are also subclinical indicators of future HF. Inflammation is strictly correlated with clotting activation, but the association between inflammation, hypercoagulability and prognosis in HF has not been previously reported. METHODS AND RESULTS: Markers of inflammation (interleukin-6; IL-6, and C-reactive protein; CRP) and hypercoagulability (D-dimer; DD, and thrombin-antithrombin III complex; TAT) were prospectively assessed in 214 subjects with New York Heart Association (NYHA) functional class II-IV HF. During a median follow-up of 8.5 months, 32 patients had an event: 13 died and 19 were hospitalized because of worsening of HF. IL-6, DD and TAT levels were all significantly associated with increased risk of death after adjustment for other known HF prognostic factors (age, gender, traditional cardiovascular risk factors, NYHA class, systolic left ventricular function, renal failure, hemoglobin, serum sodium) in a Cox multivariate proportional hazard model (P = 0.003, P = 0.01 and P = 0.02, respectively). When these markers were added simultaneously to the known prognostic factors in a new Cox multivariate model, only DD levels were significant predictors of mortality (hazard ratio [95% confidence interval; CI]: 11 [2.7-45.1], P = 0.001). The Kaplan-Meier curve revealed a significantly better outcome in patients with DD below 450 ng mL(-1). NT-pro-BNP was the only significant predictor of rehospitalization (HR [95% CI]: 5.3 [2.0-13.8], P < 0.001). CONCLUSION: Hypercoagulability and inflammation, as assessed by DD, TAT and IL-6 levels, are associated with an increased mortality risk in HF. PMID- 16689754 TI - Platelet-activating factor-acetylhydrolase A379V (exon 11) gene polymorphism is an independent and functional risk factor for premature myocardial infarction. AB - BACKGROUND: Oxidation of low density lipoproteins is an initial step of atherogenesis that generates pro-inflammatory phospholipids, including platelet activating factor (PAF). PAF is degraded by PAF-acetylhydrolase (PAF-AH), which has been postulated to be a risk factor for myocardial infarction (MI). The role of PAF-AH for the onset of premature MI is unclear. METHODS: Polymorphisms located in putatively functional regions were investigated in a cohort of patients having premature MI onset prior to 46 years of age (n = 200) and a sex age-matched control group (n = 200). The activity of PAF-AH and coronary angiograms were evaluated for the severity of coronary atherosclerosis. RESULTS: The V allele of A379V (exon 11) polymorphism on PAF-AH gene was more frequent in patients with premature MI (P = 0.001). This V allele polymorphism was also associated with a lower activity of plasma PAF-AH and a more complex coronary atherosclerosis (p Trends <0.05). Multiple logistic regression analysis showed that this polymorphism was an independent risk factor (Odds Ratio [OR] 1.66, 95% CI 1.14.1 to 5.80, P = 0.008) as well as smoking (OR 3.72, 95% CI 1.77 to 9.28, P = 0.001), diabetes mellitus (OR 2.25, 95% CI 1.40 to 5.32, P = 0.007) and hypertension (OR 1.88, 95% CI 1.25 to 5.36, P = 0.003) for the onset of premature MI. CONCLUSION: We conclude that a functional and significant association between the A379V polymorphism on exon 11 of PAF-AH gene and premature MI exists in this Taiwanese population. This polymorphism is significantly associated with the PAF AH activity and the severity of coronary atherosclerosis. PMID- 16689755 TI - Statins, fenofibrate, and quinapril increase clot permeability and enhance fibrinolysis in patients with coronary artery disease. AB - BACKGROUND: Aspirin increases fibrin clot porosity and susceptibility to lysis. It is unknown whether other drugs, in combination with aspirin, used in the treatment of coronary artery disease (CAD) might affect clot structure and resistance to lysis. AIM: The aim of the study was to assess the effects of statins, fibrates, or angiotensin-converting enzyme inhibitors (ACEIs) on fibrin clot properties. PATIENTS AND METHODS: In a randomized double-blind study, men with advanced CAD taking low-dose aspirin were assigned to receive one of the four drugs: simvastatin 40 mg day(-1) (n = 13), atorvastatin 40 mg day(-1) (n = 12), fenofibrate 160 mg day(-1) (n = 12), and quinapril 10 mg day(-1) (n = 11) for 28 +/- 2 days. Moreover, CAD patients (n = 13) taking aspirin (75 mg day(-1)) for 8 weeks were studied after additional 4 weeks on an open-label basis. Thirty men served as healthy controls. Plasma clot permeability and tissue plasminogen activator-induced fibrinolysis were evaluated at baseline and after drug administration. RESULTS: Permeability increased following the administration of simvastatin (by 20%; P = 0.01), atorvastatin (by 22%; P = 0.001), fenofibrate (by 16%; P = 0.02), and quinapril (by 13%; P = 0.04) like for aspirin (P < 0.001). Turbidity analysis showed that administration of any of the drugs was associated with higher maximum absorbancy, suggesting thicker fibers, and shorter fibrinolysis time (P < 0.001). Post-treatment reduction in lysis time correlated with an increase in clot porosity in all the groups (r from 0.42 to 0.61; P from 0.01 to 0.001). CONCLUSIONS: Statins, fibrates, and ACEIs may increase plasma clot permeability and susceptibility to fibrinolysis in CAD patients receiving aspirin. This novel antithrombotic mechanism might contribute to clinical benefits of the drugs tested. PMID- 16689756 TI - Management of primary care patients with suspected deep vein thrombosis: use of a therapeutic dose of low-molecular-weight heparin to avoid urgent ultrasonographic evaluation. AB - BACKGROUND: Out of hospital management of patients with suspected deep vein thrombosis (DVT) can be problematic. The accuracy of clinical prediction rules in the primary care setting may be inadequate, D-dimer testing may not be available, and the cost-effectiveness of urgent ultrasonographic evaluation is uncertain. OBJECTIVE: The purpose of this study was to determine the efficacy and safety of an empiric single therapeutic dose of low-molecular weight heparin (LMWH) in the time interval preceding ultrasound investigation in patients presenting to primary care physicians (PCPs) for suspicion of DVT. METHODS: Consecutive patients with suspected DVT who presented to the office of a PCP outside regular thrombosis center working hours were enrolled. All eligible patients received a single therapeutic dose of LMWH (100 anti-Xa IU kg(-1) weight) and were scheduled to undergo clinical and instrumental evaluation at the thrombosis center the morning after. Clinical events were documented after a 3-month follow-up. RESULTS: A total of 534 consecutive patients with suspected DVT were included in this study; of these 102 patients had subsequent diagnosis of DVT. We detected no episodes of pulmonary embolism, major bleeding, or death during the 18-h window between the administration of LMWH and objective evaluation. Of the 432 patients in whom diagnosis of DVT was subsequently excluded, only three (0.7%; CI: 0.2 2.0%) developed venous thromboembolic events during the 3-month follow-up period. CONCLUSIONS: Empiric treatment with a single therapeutic dose of LMWH is effective and safe for outpatients with suspected DVT initially managed in a primary care setting. This strategy has the potential to reduce the need for urgent diagnostic imaging. PMID- 16689757 TI - Accuracy of clinical decision rule, D-dimer and spiral computed tomography in patients with malignancy, previous venous thromboembolism, COPD or heart failure and in older patients with suspected pulmonary embolism. AB - BACKGROUND: The diagnostic work-up of patients with suspected pulmonary embolism (PE) has been optimized and simplified by the use of clinical decision rules (CDR), D-dimer (DD) testing and spiral computed tomography (s-CT). Whether this strategy is equally safe and efficient in specific subgroups of patients is evaluated in this study. METHODS: A diagnostic strategy including a CDR, DD test and s-CT was evaluated in patients with malignancy, previous venous thromboembolism (VTE), chronic obstructive pulmonary disease or heart failure and in older patients. PE was ruled out by either an unlikely CDR and a normal DD or a s-CT negative for PE. The safety of these tests was assessed by the 3-month incidence rate of symptomatic VTE in those without PE at baseline. The efficiency was evaluated by calculating the numbers needed to test for the different subgroups. RESULTS: The venous thromboembolic incidence rate after the combination of an unlikely CDR and a normal DD varied from 0% (95% CI: 0-7.9%) in the 482 patients older than 75 years of age to 2% (95% CI: 0.05-10.9%) in the 474 patients with a malignancy. For s-CT these incidences varied from 0.3% to 1.8%. The number needed to test in order to rule out one patient from PE with the studied strategy was highest in cancer patients and in the elderly patients (approximately 10). CONCLUSION: It appears to be safe to rule out PE by either the combination of an unlikely CDR and a normal DD or by a negative s-CT in various subgroups of patients with suspected PE. However, the clinical usefulness of the CDR in combination with the DD as the initial step in the diagnostic process varied among these patient groups. PMID- 16689758 TI - Variable region heavy chain glycosylation determines the anticoagulant activity of a factor VIII antibody. AB - BACKGROUND: N-glycosylation occurs in the variable region of about 10% of antibodies but the role of carbohydrate at this location is still poorly understood. OBJECTIVES: We investigated the function of N-glycosylation in the variable region of the heavy chain of a human monoclonal antibody, mAb-LE2E9, that partially inhibits factor VIII (FVIII) activity during coagulation. METHODS AND RESULTS: Enzymatic deglycosylation indicated that the oligosaccharides do not determine the affinity of the antibody but enhance its FVIII neutralizing activity. A mutant antibody lacking the N-glycosylation site in the variable region of the heavy chain inhibited FVIII activity by up to 40%, while inhibition by the native antibody was 80%. To evaluate the physiological effect of such a FVIII inhibition, we investigated the ability of the mutant antibody devoid of N glycosylation in the variable region to prevent thrombosis in mice with a strong prothombotic phenotype resulting from a type II deficiency mutation in the heparin binding site of antithrombin. Despite its moderate inhibition of FVIII activity, the mutant antibody significantly prevented thrombosis in treated animals. We also carried out glycan analysis of native and mutant antibodies. CONCLUSIONS: Modification of glycosylation in the variable region of antibodies contributes to the diversity of FVIII type II inhibition possibly by steric hindrance of the active site of FVIII by glycans, and may provide a novel strategy to modulate the functional activity of therapeutic antibodies. PMID- 16689759 TI - Experimental metastasis and primary tumor growth in mice with hemophilia A. AB - During experimental lung metastasis, tumor cells adhere to the pulmonary microvasculature and activate coagulation via surface-expressed tissue factor (TF), leading to local fibrin deposition and platelet aggregation. While interventional studies have demonstrated great efficacy of anticoagulants and antiplatelet agents in inhibiting metastasis, no information is available on how tumor biology may be affected by congenital bleeding disorders such as hemophilia A. We therefore used a syngeneic model to study experimental metastasis and primary tumor growth in factor VIII (FVIII)-deficient mice. By conventional reverse transcription-polymerase chain reaction, flow cytometry, and one-stage clotting assays, we demonstrated constitutive expression of TF mRNA, antigen, and procoagulant activity in the murine B16F10 melanoma cell line. In hemophilic mice, B16F10 lung metastasis was significantly (P < 0.001) enhanced by a single dose of human FVIII (100 U kg(-1)), suggesting that FVIII played a critical role during the early blood-borne phase of the metastatic cascade. In contrast, lung seeding was significantly (P < 0.05) reduced by lepirudin, a direct thrombin inhibitor, suggesting that thrombin generation contributed to pulmonary metastasis even in the absence of FVIII. Consistent with this finding, intravenous injection of B16F10 cell-evoked laboratory changes of a hemolytic thrombotic microangiopathy and consumptive coagulopathy in both hemophilic and non-hemophilic mice. Subcutaneous implantation of B16F10 cells into mice with hemophilia A gave rise to primary tumors in an exponential growth pattern similar to that observed in non-hemophilic mice. Although TF expression by B16F10 cells may promote thrombin-dependent metastasis in mice with hemophilia A, amplification of coagulation by host FVIII appears to be necessary for maximum lung seeding. PMID- 16689760 TI - Increased ADAMTS-13 proteolytic activity in rat hepatic stellate cells upon activation in vitro and in vivo. AB - INTRODUCTION: ADAMTS-13 is a member of A Disintegrin And Metalloprotease with ThromboSpondin type 1 repeats (ADAMTS) family, primarily synthesized in hepatic stellate cells (HSCs), one of the major cell types transdifferentiating into myofibroblasts during liver fibrosis. However, the association between ADAMTS-13 expression and HSC activation or liver fibrosis is not known. METHODS: In this study, we determined the ADAMTS-13 mRNA, protein, and activity in isolated primary HSCs upon activation on a plastic dish and in liver after administration of carbon tetrachloride (CCl(4)) in rats. RESULTS: We showed that ADAMTS-13 antigen and proteolytic activity in the activated rat HSCs were dramatically increased, whereas ADAMTS-13 mRNA in these cells was only minimally altered. Similarly, the ADAMTS-13 antigen and proteolytic activity in rat liver after CCl(4) injury were also significantly increased, whereas the ADAMTS-13 mRNAs in these liver tissues were only slightly increased compared with normal. Surprisingly, despite the dramatic up-regulation of ADAMTS-13 protein synthesis in the activated HSCs after CCl(4) administration, the plasma levels of ADAMTS-13 protease in rats did not increase concordantly. CONCLUSION: We conclude that the up-regulation of ADAMTS-13 protein expression in rat HSCs during activation in vitro and in vivo suggests the possibility of ADAMTS-13 proteolysis, an important part of function of the activated HSCs, perhaps through modulation of liver regeneration or formation of liver fibrosis after various injuries. The data also suggest the minimal contribution of the activated HSCs in regulation of plasma levels of ADAMTS-13 protease. PMID- 16689761 TI - Thrombin generation in factor VIII-depleted neonatal plasma: nearly normal because of physiologically low antithrombin and tissue factor pathway inhibitor. AB - BACKGROUND: Bleeding in hemophilic neonates has a low incidence. A possible explanation for this could be the peculiarities of the neonatal hemostatic system, especially low levels of the inhibitors tissue factor pathway inhibitor (TFPI) and antithrombin (AT). OBJECTIVE: We investigated the influence of an elevation of these inhibitors to adult levels on the thrombin generation (TG) in normal neonatal plasma and factor (F) VIII-depleted neonatal plasma by means of incubation with anti-FVIII-antibodies. PATIENTS/METHODS: TG was measured after activation with low amounts of tissue factor (TF) by using Calibrated Automated Thrombography. RESULTS: TG in FVIII-depleted neonatal plasma was nearly as high as in normal neonatal plasma. TG decreased after elevation of AT in both neonatal plasmas. After elevation of TFPI TG decreased much more in FVIII-depleted neonatal plasma than in normal neonatal plasma. After elevation of both inhibitors their synergistic effect led to a stronger decrease of TG in FVIII depleted neonatal plasma. TG measured in plasma of one hemophilic newborn showed the same pattern as in FVIII-depleted neonatal plasma. CONCLUSION: Our observation provides a biochemical basis for the rare bleeding in hemophilic neonates and shows the important role of the natural inhibitors in the hemostatic system of hemophilic patients. PMID- 16689762 TI - The prothrombin 20209 C-->T mutation in Jewish-Moroccan Caucasians: molecular analysis of gain-of-function of 3' end processing. AB - BACKGROUND: Mutations of the 3' end mRNA-processing signal of the prothrombin (F2) gene have been reported to cause elevated F2 plasma concentrations, thrombosis, and complications of pregnancy. Whereas the common F2 20210*A mutation is almost exclusively found in Caucasians, the F2 20209*T mutation has been reported in Afro-Americans and Afro-Caribbeans only. PATIENTS AND METHODS: Using LightCycler technology, three unrelated Jewish-Moroccan patients tested for obstetric complications were found to be carriers of the F2 20209*T allele. A detailed molecular analysis was performed to identify the functional impact of this mutation. RESULTS: We report three unrelated women of Jewish-Moroccan origin with a F2 20209*T mutation and fetal loss or infertility. The functional analysis revealed that the F2 20209*T mutation stimulates 3' end processing and up regulates prothrombin protein expression as assessed by a highly sensitive luminescence-based reporter system. CONCLUSIONS: This is the first report of 20209*T in Caucasians, and functional analysis demonstrates that F2 20209*T falls into a general category of mutations of the F2 gene, which may possibly contribute to thrombophilia and complications of pregnancy by interfering with a tightly balanced architecture of non-canonical F2 3' end formation signals. PMID- 16689763 TI - Insights into abnormal hemostasis in the Quebec platelet disorder from analyses of clot lysis. AB - BACKGROUND: The Quebec platelet disorder (QPD) is inherited and characterized by delayed-onset bleeding following hemostatic challenge. Other characteristics include increased expression and storage of active urokinase-type plasminogen activator (u-PA) in platelets in the setting of normal to increased u-PA in plasma. There is also consumption of platelet plasminogen activator inhibitor-1 and increased generation of plasmin in platelets accompanied by proteolysis of stored alpha-granule proteins, including Factor V. AIMS AND METHODS: Although fibrinolysis has been proposed to contribute to QPD bleeding, the effects of QPD blood and platelets on clot lysis have not been evaluated. We used thromboelastography (TEG), biochemical evaluations of whole blood clot lysis, assessments of clot ultrastructure, and perfusion of blood over preformed fibrin to gain insights into the disturbed hemostasis in the QPD. RESULTS: Thromboelastography was not sensitive to the increased u-PA in QPD blood. However, there was abnormal plasmin generation in QPD whole blood clots, generated at low shear, with biochemical evidence of increased fibrinolysis. The incorporation of QPD platelets into a forming clot led to progressive disruption of fibrin and platelet aggregates unless drugs were added to inhibit plasmin. In whole blood perfusion studies, QPD platelets showed normal adherence to fibrin, but their adhesion was followed by accelerated fibrinolysis. CONCLUSIONS: The QPD is associated with "gain-of-function" abnormalities that increase the lysis of forming or preformed clots. These findings suggest accelerated fibrinolysis is an important contributor to QPD bleeding. PMID- 16689764 TI - Generation of platelet angiostatin mediated by urokinase plasminogen activator: effects on angiogenesis. AB - BACKGROUND: Angiogenesis, the growth of new capillaries from pre-existing blood vessels, is regulated by a balance between its promoters and inhibitors. Platelets are an important circulating store of angiogenesis regulators. We have previously identified the angiogenesis inhibitor angiostatin in human platelets. AIM: To identify the mechanism of platelet angiostatin generation and its pharmacological regulation. METHODS: Platelet aggregometry, flow cytometry, Western blot, zymography, immunofluorescence microscopy, matrigel-induced angiogenesis of human umbilical vein endothelial cells (HUVECs), and a panel of selective proteinase inhibitors were used to study the mechanism of angiostatin generation by platelets, its pharmacological regulation, and effects on angiogenesis. Release of pro-MMP-2 by HUVECs was also used to quantify angiogenesis. RESULTS: Platelet membranes were identified as the site of angiostatin generation from plasminogen. Generation of angiostatin by platelet membranes was not affected by a matrix metalloproteinase (MMP) inhibitor, phenanthroline, but was inhibited by serine proteinase inhibitors aprotinin, leupeptin, plasminogen activator inhibitor-1, and selective inhibitor of urokinase plasminogen activator (uPA), uPA-STOP(TM). Angiostatin generation by intact platelets was inhibited by aprotinin, and the resulting incubate promoted angiogenesis to a greater extent than incubate where angiostatin generation occurred. Furthermore, HUVECs incubated with reaction mixture, where angiostatin generation was inhibited, released more pro-MMP-2 than HUVECs incubated with supernatants, where angiostatin generation occurred. CONCLUSIONS: We conclude that; (i) platelets constitutively generate angiostatin on their membranes; (ii) this mechanism is dependent on uPA, but not, MMPs; and (iii) inhibition of platelet angiostatin generation can further promote angiogenesis. PMID- 16689765 TI - Effect of multimer size and a natural dimorphism on the binding of convulxin to platelet glycoprotein (GP)VI. AB - BACKGROUND: Convulxin (CVX), a C-type lectin from the venom of Crotalus durissus terrificus, is a potent activator of human platelets, binding predominantly to glycoprotein (GP)VI. Native CVX is an octamer composed of four alphabeta heterodimers [(alphabeta)(4)]. Two different native sequences have been reported, one bearing lysine (K), the other glutamic acid (E), at beta chain residue 89, but the physiological relevance of this difference is unknown. OBJECTIVE: We used the Drosophila S2 system to express recombinant CVX (rCVX) heterodimers (alphabeta) and site-directed mutagenesis to evaluate the influence of multimer size and the substitution betaK89E on CVX function. METHODS: By flow cytometry, native CVX and both recombinant forms bind to human platelets in whole blood. By surface plasmon resonance (BIAcore, Piscataway, NJ, USA), the calculated equilibrium dissociation constants (K(D)) were: rCVX alphabeta89K, 11.3 x 10(-8) m; rCVX alphabeta89E, 9 x 10(-8) m; and native CVX, 2.8 x 10(-8) m. RESULTS: Thus, the affinities of the two rCVX forms for human, recombinant GPVI are essentially the same, but the relative affinity of native CVX is about 3-fold higher. The minimum concentration of native CVX that induces maximal human platelet aggregation (70 pm) is roughly 400-fold lower than that of either rCVX (29 nm). CONCLUSIONS: These results are consistent with the hypothesis that the ability of the native CVX octamer to cluster mobile GPVI molecules within the platelet membrane may be the single most important factor that contributes to the efficiency with which CVX is able to induce platelet activation. PMID- 16689767 TI - The role of the fibrocyte in intimal hyperplasia. AB - BACKGROUND: Experimental animal studies have shown that the intimal hyperplasia (IH) responsible for occlusion after successful revascularization procedures may be partially caused by a bone marrow-derived cell that migrates to the site of vascular injury. Concurrent studies have demonstrated an extensive role in wound healing for the circulating fibrocyte. OBJECTIVES: We aimed to trace the path of the circulating cell that contributes to IH and determine if it is the fibrocyte. METHODS AND RESULTS: We established an in vitro model whereby purified monocytes from six healthy human volunteers were cultured into fibrocytes. These cells were morphometrically similar to the vascular smooth muscle cell (VSMC) found in IH and expressed alpha-smooth muscle actin (alpha-SMA) as well as CD34, CD45 and Collagen I (Col I), markers indicative of the fibrocyte. In an in vivo ovine carotid artery synthetic patch graft model, carboxyfluorescein diacetate, succinimidyl ester (CFSE) labeled circulating leukocytes were observed throughout the graft as well as in the neointima in 18 sheep. These cells were shown to produce collagen and alpha-SMA at 1, 2 and 4 weeks. These cells then underwent immunohistochemical analysis and were found to express a set of markers unique to the fibrocyte (CD34, CD45, Vimentin and alpha-SMA) and also to double stain for CD34 and alpha-SMA. CONCLUSIONS: IH in an ovine carotid artery patch graft model is partially derived from a hematopoietic circulating progenitor cell that acquires mesenchymal features as it matures at the site of injury. PMID- 16689766 TI - A GPI-anchored co-receptor for tissue factor pathway inhibitor controls its intracellular trafficking and cell surface expression. AB - BACKGROUND: Tissue factor pathway inhibitor (TFPI) lacks a membrane attachment signal but it remains associated with the endothelial surface via its association with an, as yet, unidentified glycosyl phosphatidylinositol (GPI)-anchored co receptor. OBJECTIVES/METHODS: Cellular trafficking of TFPI within aerolysin resistant ECV304 and EA.hy926 cells, which do not express GPI-anchored proteins on their surface, was compared with their wild-type counterparts. RESULTS AND CONCLUSIONS: Although aerolysin-resistant cells produce normal amounts of TFPI mRNA, TFPI is not expressed on the cell surface and total cellular TFPI is greatly decreased compared with wild-type cells. Additionally, normal, not increased, amounts of TFPI are secreted into conditioned media indicating that TFPI is degraded within the aerolysin-resistant cells. Confocal microscopy and studies using metabolic inhibitors demonstrate that aerolysin-resistant cells produce TFPI and transport it into the Golgi with subsequent degradation in lysosomes. The experimental results provide no evidence that cell surface TFPI originates from secreted TFPI that binds back to a GPI-anchored protein. Instead, the data suggest that TFPI tightly, but reversibly, binds to a GPI anchored co receptor in the ER/Golgi. The co-receptor then acts as a molecular chaperone for TFPI by trafficking it to the cell surface of wild-type cells or to lysosomes of aerolysin-resistant cells. TFPI that escapes co-receptor binding is secreted through the same pathway in both wild-type and aerolysin-resistant cells. The data provide a framework for understanding how TFPI is expressed on endothelium. PMID- 16689768 TI - Fibrinogen Paris IX: a case of symptomatic hypofibrinogenemia with Bbeta Y236C and Bbeta IVS7-1G-->C mutations. PMID- 16689769 TI - Determination of serotonin release from platelets by HPLC and ELISA in the diagnosis of heparin-induced thrombocytopenia: comparison with reference method by [C]-serotonin release assay. PMID- 16689770 TI - The structure of fibrinogen fragment D with the 'A' knob peptide GPRVVE. PMID- 16689771 TI - Factor VIII intron-1 inversion: frequency and inhibitor prevalence. PMID- 16689772 TI - Clopidogrel resistance caused by a failure to metabolize clopidogrel into its metabolites. PMID- 16689773 TI - Measurement of ADAMTS-13 activity in plasma by the FRETS-VWF73 assay: comparison with other assay methods. PMID- 16689774 TI - The role of pulse dexamethasone in acquired idiopathic thrombotic thrombocytopenic purpura. PMID- 16689775 TI - Is adiponectin implicated in venous thromboembolism? PMID- 16689776 TI - Severe type I protein C deficiency with neonatal purpura fulminans due to a novel homozygous mutation in exon 6 of the protein C gene. PMID- 16689777 TI - Molecular mechanism for hereditary protein C deficiency in two Chinese families with thrombosis. PMID- 16689778 TI - Alcohol, cardiovascular risk, and health: there is a window for benefits. PMID- 16689780 TI - A rapid genotyping method for the vitamin K epoxide reductase complex subunit 1 (VKORC1) gene. PMID- 16689781 TI - Elimination of contact factor activation improves measurement of platelet dependent thrombin generation by calibrated automated thrombography at low concentration tissue factor. PMID- 16689782 TI - An evaluation of cyclosporine alone for the treatment of early recurrences of thombotic thrombocytopenic purpura. PMID- 16689783 TI - New International Diabetes Federation (IDF) and National Cholesterol Education Program Adult Treatment panel III (NCEP-ATPIII) criteria and the involvement of hemostasis and fibrinolysis in the metabolic syndrome. PMID- 16689784 TI - More on: are randomized clinical trials the only truth? Not always. PMID- 16689786 TI - Cell wall assembly in Bacillus subtilis: how spirals and spaces challenge paradigms. AB - Although the bacterial cell wall has been the subject of decades of investigation, recent studies continue to generate novel and controversial models of its synthesis and assembly. Here we compare and contrast the transcompartmental biosyntheses of peptidoglycan and teichoic acid in Bacillus subtilis. In addition, the current paradigms of B. subtilis wall assembly and structure are distinguished from emerging models of murein insertion and organization. We discuss evidence for the directed, cytoskeleton-dependent insertion of nascent peptidoglycan and the existence of a periplasmic compartment. Furthermore, we summarize the challenges these findings represent to the existing paradigm of murein insertion. Finally, motivated by these new developments, we discuss outstanding issues that remain to be addressed and suggest research directions that may contribute to a better understanding of cell wall assembly in B. subtilis. PMID- 16689787 TI - Poly-gamma-glutamate in bacteria. AB - Poly-gamma-glutamate (PGA), a natural polymer, is synthesized by several bacteria (all Gram-positive), one archaea and one eukaryote. PGA has diverse biochemical properties, enabling it to play different roles, depending on the organism and its environment. Indeed, PGA allows bacteria to survive at high salt concentrations and may also be involved in virulence. The minimal gene sets required for PGA synthesis were recently defined. There are currently two nomenclatures depending on the PGA final status: cap, for 'capsule', when PGA is surface associated or pgs, for 'polyglutamate synthase', when PGA is released. The minimal gene sets contain four genes termed cap or pgs B, C, A and E. The PGA synthesis complex is membrane-anchored and uses glutamate and ATP as substrates. Schematically, the reaction may be divided into two steps, PGA synthesis and PGA transport through the membrane. PGA synthesis depends primarily on CapB-CapC (or PgsB-PgsC), whereas PGA transport requires the presence, or the addition, of CapA CapE (or PgsAA-PgsE). The synthesis complex is probably responsible for the stereochemical specificity of PGA composition. Finally, PGA may be anchored to the bacterial surface or released. An additional enzyme is involved in this reaction: either CapD, a gamma-glutamyl-transpeptidase that catalyses anchorage of the PGA, or PgsS, a hydrolase that facilitates release. The anchoring of PGA to the bacterial surface is important for virulence. All cap genes are therefore potential targets for inhibitors specifically blocking PGA synthesis or anchorage. PMID- 16689788 TI - The cellular response to chromosome breakage. AB - DNA double-strand breaks (DSBs) are among the most deleterious types of damage that can occur in the genome of eukaryotic cells because failure to repair them can lead to loss of genetic information and chromosome rearrangements. DSBs can arise by failures in DNA replication and by exposure to environmental factors, such as ionizing radiations and radiomimetic chemicals. Moreover, they might arise when telomeres undergo extensive erosion, leading to the activation of the DNA damage response pathways and the onset of apoptosis and/or senescence. Importantly, DSBs can also form in a programmed manner during development. For example, meiotic recombination and rearrangement of the immunoglobulin genes in lymphocytes require the generation of site- or region-specific DSBs through the action of specific endonucleases. Efficient DSB repair is crucial in safeguarding genome integrity, whose maintenance in the face of DSBs involves branched signalling networks that switch on DNA damage checkpoints, activate DNA repair, induce chromatin reorganization and modulate numerous cellular processes. Not surprisingly, defects in these networks result in a variety of diseases ranging from severe genetic disorders to cancer predisposition and accelerated ageing. PMID- 16689789 TI - Role of the methylcitrate cycle in Mycobacterium tuberculosis metabolism, intracellular growth, and virulence. AB - Growth of bacteria and fungi on fatty acid substrates requires the catabolic beta oxidation cycle and the anaplerotic glyoxylate cycle. Propionyl-CoA generated by beta-oxidation of odd-chain fatty acids is metabolized via the methylcitrate cycle. Mycobacterium tuberculosis possesses homologues of methylcitrate synthase (MCS) and methylcitrate dehydratase (MCD) but not 2-methylisocitrate lyase (MCL). Although MCLs share limited homology with isocitrate lyases (ICLs) of the glyoxylate cycle, these enzymes are thought to be functionally non-overlapping. Previously we reported that the M. tuberculosis ICL isoforms 1 and 2 are jointly required for growth on fatty acids, in macrophages, and in mice. ICL-deficient bacteria could not grow on propionate, suggesting that in M. tuberculosis ICL1 and ICL2 might function as ICLs in the glyoxylate cycle and as MCLs in the methylcitrate cycle. Here we provide biochemical and genetic evidence supporting this interpretation. The role of the methylcitrate cycle in M. tuberculosis metabolism was further evaluated by constructing a mutant strain in which prpC (encoding MCS) and prpD (encoding MCD) were deleted. The DeltaprpDC strain could not grow on propionate media in vitro or in murine bone marrow-derived macrophages infected ex vivo; growth under these conditions was restored by complementation with a plasmid containing prpDC. Paradoxically, bacterial growth and persistence, and tissue pathology, were indistinguishable in mice infected with wild-type or DeltaprpDC bacteria. PMID- 16689790 TI - Evolution of bacteriophages infecting encapsulated bacteria: lessons from Escherichia coli K1-specific phages. AB - Bacterial capsules are not only important virulence factors, but also provide attachment sites for bacteriophages that possess capsule degrading enzymes as tailspike proteins. To gain insight into the evolution of these specialized viruses, we studied a panel of tailed phages specific for Escherichia coli K1, a neuroinvasive pathogen with a polysialic acid capsule. Genome sequencing of two lytic K1-phages and comparative analyses including a K1-prophage revealed that K1 phages did not evolve from a common ancestor. By contrast, each phage is related to a different progenitor type, namely T7-, SP6-, and P22-like phages, and gained new host specificity by horizontal uptake of an endosialidase gene. The new tailspikes emerged by combining endosialidase domains with the capsid binding module of the respective ancestor. For SP6-like phages, we identified a degenerated tailspike protein which now acts as versatile adaptor protein interconnecting tail and newly acquired tailspikes and demonstrate that this adapter utilizes an N-terminal undecapeptide interface to bind otherwise unrelated tailspikes. Combining biochemical and sequence analyses with available structural data, we provide new molecular insight into basic mechanisms that allow changes in host specificity while a conserved head and tail architecture is maintained. Thereby, the present study contributes not only to an improved understanding of phage evolution and host-range extension but may also facilitate the on purpose design of therapeutic phages based on well-characterized template phages. PMID- 16689791 TI - Sex and virulence in Escherichia coli: an evolutionary perspective. AB - Pathogenic Escherichia coli cause over 160 million cases of dysentery and one million deaths per year, whereas non-pathogenic E. coli constitute part of the normal intestinal flora of healthy mammals and birds. The evolutionary pathways underlying this dichotomy in bacterial lifestyle were investigated by multilocus sequence typing of a global collection of isolates. Specific pathogen types [enterohaemorrhagic E. coli, enteropathogenic E. coli, enteroinvasive E. coli, K1 and Shigella] have arisen independently and repeatedly in several lineages, whereas other lineages contain only few pathogens. Rates of evolution have accelerated in pathogenic lineages, culminating in highly virulent organisms whose genomic contents are altered frequently by increased rates of homologous recombination; thus, the evolution of virulence is linked to bacterial sex. This long-term pattern of evolution was observed in genes distributed throughout the genome, and thereby is the likely result of episodic selection for strains that can escape the host immune response. PMID- 16689792 TI - M. tuberculosis Rv2252 encodes a diacylglycerol kinase involved in the biosynthesis of phosphatidylinositol mannosides (PIMs). AB - Phosphorylated lipids play important roles in biological systems, not only as structural moieties but also as modulators of cellular function. Phospholipids of pathogenic bacteria are known to play roles both as membrane components and as factors that modulate the infectious process. Mycobacterium tuberculosis is, however, noteworthy in that it has an extremely diverse repertoire of biologically active phosphorylated lipids that, in the absence of a specialized protein translocation system, appear to constitute the main means of communication with the host. Many of these lipids are derived from phosphatidylinositol (PI) that is differentially processed to give rise to phosphatidylinositol mannosides (PIMs) or lipoarabinomannan. In preliminary studies on the lipid processing enzymes associated with the bacterial cell wall, a kinase activity was noted that gave rise to a novel lipid species released by the bacterium. It was determined that this kinase activity was encoded by the ORF Rv2252. Rv2252 demonstrates the capacity to phosphorylate various amphipathic lipids of host and bacterial origin, in particular a M. tuberculosis derived diacylglycerol. Targeted deletion of the rv2252 gene resulted in disruption of the production of certain higher order PIM species, suggesting a role for Rv2252 in the biosynthetic pathway of PI, a PIM precursor. PMID- 16689793 TI - Fine-scale time-lapse analysis of the biphasic, dynamic behaviour of the two Vibrio cholerae chromosomes. AB - Using fluorescent repressor-operator systems in live cells, we investigated the dynamic behaviour of chromosomal origins in Vibrio cholerae, whose genome is divided between two chromosomes. We have developed a method of analysing fine scale motion in the curved co-ordinate system of vibrioid bacteria. Using this method, we characterized two different modes of chromosome behaviour corresponding to periods between segregation events and periods of segregation. Between segregation events, the origin positions are not fixed but rather maintained within ellipsoidal caged domains, similar to eukaryotic interphase chromosome territories. These domains are approximately 0.4 microm wide and 0.6 microm long, reflecting greater restriction in the short axis of the cell. During segregation, movement is directionally biased, speed is comparable between origins, and cell growth can account for nearly 20% of the motion observed. Furthermore, the home domain of each origin is positioned by a different mechanism. Specifically, the oriC(I) domain is maintained at a constant actual distance from the pole regardless of cell length, while the oriC(II) domain is maintained at a constant relative position. Thus the actual position of oriC(II) varies with cell length. While the gross behaviours of the two origins are distinct, their fine-scale dynamics are remarkably similar, indicating that both experience similar microenvironments. PMID- 16689794 TI - Crystal structure of the DNA-binding domain of BldD, a central regulator of aerial mycelium formation in Streptomyces coelicolor A3(2). AB - BldD is a central regulator of the developmental process in Streptomyces coelicolor. The 1.8 angstroms resolution structure of the DNA-binding domain of BldD (BldDN) reveals that BldDN forms a compact globular domain composed of four helices (alpha1-alpha4) containing a helix-turn-helix motif (alpha2-alpha3) resembling that of the DNA-binding domain of lambda repressor. The BldDN/DNA complex model led us to design a series of mutants, which revealed the important role of alpha3 and the 'turn' region between alpha2 and alpha3 for DNA recognition. Based on the fact that BldD occupies two operator sites of bldN and whiG and shows significant disparity in the affinity toward the two operator sites when they are disconnected, we propose a model of cooperative binding, which means that the binding of one BldD dimer to the high affinity site facilitates that of the second BldD dimer to the low affinity site. In addition, structural and mutational investigation reveals that the Tyr62Cys mutation, found in the first-identified bldD mutant, can destabilize BldD structure by disrupting the hydrophobic core. PMID- 16689795 TI - Innate immune responses regulate trypanosome parasite infection of the tsetse fly Glossina morsitans morsitans. AB - Tsetse flies transmit the protozoan parasite African trypanosomes, the agents of human sleeping sickness in sub-Saharan Africa. Parasite transmission in the insect is restricted by a natural resistance phenomenon (refractoriness). Understanding the mechanism of parasite resistance is important as strengthening fly's response(s) via transgenic approaches can prevent parasite transmission and lead to the development of novel vector control strategies. Here, we investigated the role of one of the two major pathways regulating innate immunity in invertebrates, the immunodeficiency (Imd) pathway, for Glossina morsitans morsitans's natural defence against Trypanosoma brucei spp. infections. We determined the molecular structure of the Imd pathway transcriptional activator Relish (GmmRel), which shows high amino acid identity and structural similarity to its Drosophila homologue. Through a double-stranded RNA-based interference approach, we showed that the pathogen-induced expression profile of the antimicrobial peptides (AMPs) attacin and cecropin is under the regulation of GmmRel. Unexpectedly, the AMP diptericin appears to be constitutively expressed in tsetse independent of the presence of the Rel factor. Through GmmRel knock down, we could successfully block the induction of attacin and cecropin expression in the immune responsive tissues fat body and proventriculus (cardia) following microbial challenge. The midgut and salivary gland trypanosome infection prevalence, as well as the intensity of midgut parasite infections were found to be significantly higher in flies when attacin and relish expression were knocked down. Our results provide the first direct evidence for the involvement of antimicrobial peptides in trypanosome transmission in tsetse. PMID- 16689796 TI - A protein Ser/Thr kinase cascade negatively regulates the DNA-binding activity of MrpC, a smaller form of which may be necessary for the Myxococcus xanthus development. AB - The developmental process of Myxococcus xanthus is achieved by the expression of a specific set of genes under the influence of developmental signals. MrpC is a member of the CRP family of transcription regulators, essential for fruA expression during development. The Pkn8-Pkn14 protein kinase cascade negatively regulates mrpC expression (H. Nariya and S. Inouye, 2005. Mol Microbiol 58: 367 379). Elevated levels of mrpC in pkn8 and pkn14 deletion strains (Deltapkn8 and Deltapkn14) induce untimely FruA production during vegetative growth resulting in significantly faster fruiting body development. mrpC expression is presumably activated by MrpA and MrpB which belong to a two-component His-Asp phosphorelay system and is proposed to require MrpC on the basis of the genetic analysis. In the present study, we demonstrate that MrpC binds to at least eight sites in the upstream region of its promoter. Based on analysis of MrpC binding sites in the mrpC and fruA promoter regions, there are two types of MrpC-specific binding sequences. Importantly, MrpC-binding activity was greatly reduced upon its phosphorylation by Pkn14. MrpC2, a transcription activator for fruA expression, lacks the N-terminal 25 residues of MrpC and exhibited four- and eightfold greater binding activity to the mrpC and fruA promoter regions respectively. Pkn14 was not able to phosphorylate MrpC2 and phosphorylates MrpC at Thr residue(s), thus Thr-21 and/or Thr-22 is (are) the likely site(s) of MrpC phosphorylation. MrpC2 was not detected in a lonD mutant in which fruA expression is low. Thus, the LonD protease essential for development may play an important role for the activation of MrpC-binding activity through its proteolytic processing to MrpC2, required for developmental progression. MrpC2, only detectable during development in DZF1, was present at high levels during vegetative growth in Deltapkn8 and Deltapkn14, thus MrpC phosphorylation may inhibit its proteolytic processing. Based on these results, we propose a mechanism by which two transcription factors essential to development, MrpC and FruA, are regulated during the M. xanthus life cycle. PMID- 16689797 TI - In vivo drafting of single-chain antibodies for regulatory duty on the sigma54 promoter Pu of the TOL plasmid. AB - The identification of single-chain antibodies (scFvs) that interfere in vivo with the building of the complex that activate the prokaryotic, sigma54-dependent promoter Pu of the catabolic TOL plasmid pWW0 is reported. To this end, a phage M13 library of scFvs was raised against the cognate prokaryotic enhancer-binding activator, XylR. The scFv pool was then expressed intracellularly in a reporter Pu-lacZ strain of Escherichia coli designed to permit formation of intramolecular disulphide bonds in cytoplasmic proteins. This strain allowed the assembly of functional scFvs and the direct testing of their activity on the Pu promoter in vivo. Specifically, genetic screening for lacZ-minus colonies yielded a number of scFvs able to downregulate transcriptional output in live cells. Two antibody clones were purified and shown to inhibit the activity of the same promoter in vitro as well. These scFvs targeted the DNA-binding domain of XylR and its ATP binding site respectively. This work provides a proof of principle that mimetic regulatory factors can be derived from an antibody repertoire that specifically interact with given transcriptional activators. As assembly of initiation complexes is stimulated or inhibited by regulatory proteins we argue that anti XylR scFvs operate as bona fide transcriptional inhibitors of the Pu promoter. PMID- 16689798 TI - Characterization of a conjugative transposon integrase, IntDOT. AB - Sequence analysis revealed that the integrase of the Bacteroides conjugative transposon CTnDOT (IntDOT) might be a member of the tyrosine recombinase family because IntDOT has five of six highly conserved residues found in the catalytic domains of tyrosine recombinases. Yet, IntDOT catalyses a reaction that appears to differ in some respects from well-studied tyrosine recombinases such as that of phage lambda. To assess the importance of the conserved residues, we changed residues in IntDOT that align with conserved residues in tyrosine recombinases. Some substitutions resulted in a complete loss or significant decrease of integration activity in vivo. The ability of the mutant proteins to cleave and ligate CTnDOT attachment site (attDOT) DNA in vitro in general paralleled the in vivo results, but the H345A mutant, which had a wild-type level of integration in vivo, exhibited a slightly lower level of cleavage and ligation in vitro. Our results confirm the hypothesis that IntDOT belongs to the tyrosine recombinase family, but the catalytic core of the protein seems to have somewhat different organization. Previous DNA sequence analyses showed that CTnDOT att sites contain 5 bp non-homologous coupling sequences which were assumed to define the putative staggered sites of cleavage. However, cleavage assays showed that one of the cleavage sites is 2 bp away from the junction of CTnDOT and coupling sequence DNA. The site is in a region of homology that is conserved in CTnDOT att sites. PMID- 16689799 TI - Genetic exchange of the S2 and S3 subunits in pertussis toxin. AB - Bordetella pertussis, the causative agent of whooping cough, produces a complex hetero-oligomeric exotoxin, named pertussis toxin (PTX), which is responsible for several of the clinical manifestations associated with whooping cough. The toxin is composed of five dissimilar subunits, named S1 through S5 and arranged in a hexameric structure with a 1S1:1S2:1S3:2S4:1S5 stoichiometry. Although S2 and S3 share 70% amino acid identity, these two subunits were previously thought not to be able to substitute for each other in toxin assembly/secretion and the biological activities of PTX. Here, we show that toxin analogues containing two S3 subunits and lacking S2 (PTXdeltaS2), or containing two S2 subunits and lacking S3 (PTXdeltaS3), can be produced, assembled and secreted by B. pertussis strains, in which the S2-encoding cistron or the S3-coding cistrons have been inactivated by internal in-frame deletions that avoid downstream effects. In fact, PTXdeltaS3 was produced in higher amounts in the bacterial culture supernatants than natural PTX, whereas PTXdeltaS2 was produced in lower amounts than PTX. The action of the toxin analogues on the clustering of Chinese Hamster Ovary cells was also affected differentially by the S2-S3 substitution. These toxin analogues constitute thus interesting probes for the study of cellular functions, in particular immune cell functions, for which natural PTX has already shown its usefulness. PMID- 16689800 TI - The effect of promoter strength, supercoiling and secondary structure on mutation rates in Escherichia coli. AB - Four mutations resulting in opal stop codons were individually engineered into a plasmid-borne chloramphenicol-resistance (cat) gene driven by the lac promoter. These four mutations were located at different sites in secondary structures. The mutations were analysed with the computer program mfg, which predicted their relative reversion frequencies. Reversion frequencies determined experimentally correlated with the mutability of the bases as predicted by mfg. To examine the effect of increased transcription on reversion frequencies, the lac promoter was replaced with the stronger tac promoter, which resulted in 12- to 30-fold increases in reversion rates. The effect of increased and decreased supercoiling was also investigated. The cat mutants had higher reversion rates in a topA mutant strain with increased negative supercoiling compared with wild-type levels, and the cat reversion rates were lower in a topA gyrB mutant strain with decreased negative supercoiling, as predicted. PMID- 16689801 TI - The Campylobacter jejuni PEB1a adhesin is an aspartate/glutamate-binding protein of an ABC transporter essential for microaerobic growth on dicarboxylic amino acids. AB - The PEB1a protein of the gastrointestinal pathogen Campylobacter jejuni mediates interactions with epithelial cells and is an important factor in host colonization. Cell fractionation and immunoblotting showed that PEB1a is most abundant in the periplasm of C. jejuni, and is detectable in the culture supernatant but not in the inner or outer membrane. The protein is homologous with periplasmic-binding proteins associated with ABC transporters and we show by fluorescence spectroscopy that purified recombinant PEB1a binds L-aspartate and L glutamate with sub microM K(d) values. Binding of L-14C-aspartate or L-14C glutamate was strongly out-competed by excess unlabelled aspartate or glutamate but only poorly by asparagine and glutamine. A mutant in the Cj0921c gene, encoding PEB1a, was completely unable to transport 5 microM L-14C-glutamate and showed a large reduction (approximately 20-fold) in the rate of L-14C-aspartate transport compared with the wild type. Although microaerobic growth of this mutant was little affected in complex media, growth on aspartate or glutamate in defined media was completely prevented, whereas growth with serine was similar to wild type. 1H-NMR analysis of the culture supernatants of the Cj0921c mutant showed some utilization of aspartate but not glutamate, consistent with the transport data. It is concluded that in addition to the established role of PEB1a as an adhesin, the PEB1 transport system plays a key role in the utilization of aspartate and glutamate, which may be important in vivo carbon sources for this pathogen. PMID- 16689802 TI - Regulated expression of glutamyl-tRNA synthetase is directed by a mobile genetic element in the cyanobacterium Tolypothrix sp. PCC 7601. AB - The genome of Tolypothrix sp. PCC 7601 carries two copies of a novel insertion sequence, ISTosp1. One of the two copies is located upstream of the gene encoding glutamyl-tRNA synthetase, an enzyme playing a key role in protein and pigment synthesis. The tnpA gene of the IS element and gltX were co-transcribed and their expression was transiently upregulated upon retrieval of the ammonium source irrespective of whether nitrate or no nitrogen source were available. The second copy is also transcribed and shows a similar regulatory pattern. Structural elements of the promoter (-10 and -35 sequences) directing the expression of the tnpA-gltX operon have been localized within the IS. Regulatory sequences involving the NtcA transcription factor in the control of tnpA-gltX expression were found both within and in sequences upstream of the insertion element. The expression of gltX in a closely related cyanobacterium, Nostoc sp. PCC 7120, which lacks the insertion upstream of gltX, decreased upon ammonium retrieval, a regulatory pattern that markedly differs from that observed in Tolypothrix sp. PCC 7601. ISTosp1 constitutes a good example of how cells can make use of a transposable element to evolve an original regulatory mechanism. PMID- 16689803 TI - Transcriptional regulation of the agr locus and the identification of DNA binding residues of the global regulatory protein SarR in Staphylococcus aureus. AB - Many members of the transcriptional regulator SarA protein family are winged helix proteins that are involved in gene regulation essential to Staphylococcus aureus pathogenesis. Investigation of the mechanism by which this family of genes acts at the molecular level will likely contribute to the understanding of the pathogenesis process and enhance the potential for the development of inhibitors capable of controlling S. aureus infections. Our previously published studies demonstrate that SarR is a repressor of sarA expression. Here, we show that SarR (a member of the SarA protein family) in part regulates agr expression by direct binding to the agr intergenic promoter region as determined by gel shift and DNase I footprinting assays. Analysis of the double sarA/sarR mutant in the early phase of growth reveals its significant role in regulating agr expression as compared with single mutants. Based on the previously reported crystal structure of SarR, we conducted site-specific mutagenesis and demonstrate that K52 residues within helix-turn-helix (HTH), K80, R82 and R88 (in the wing) and L105 (in the alpha5 helix) are important for DNA binding. Interestingly, SarR and SarA binding sites on the agr promoter are confined within the same region of DNA. Additional gel shift studies with SarR and SarA suggest that these two proteins may bind the same region of the agr promoter. PMID- 16689805 TI - Assessment of skin exposure to nickel, chromium and cobalt by acid wipe sampling and ICP-MS. AB - There is a great need to accurately assess skin exposure to contact allergens. We have developed a technique for assessment of skin exposure to nickel, chromium and cobalt using acid wipe sampling by cellulose wipes with 1% nitric acid. Chemical analysis was performed by inductively coupled plasma mass spectrometry (ICP-MS). The recovery of nickel, chromium and cobalt from arms and palms was 93%. The analytical result is expressed in terms of mass per unit area (microg/cm(2)). The developed acid wipe sampling technique is suitable for determination of nickel, chromium and cobalt deposited on the skin. The technique may be used in workplace studies, in studies of individuals in the general population, in dermatitis patients, in identification of risk groups, as well as in developing preventive strategies and in follow-up after intervention. PMID- 16689804 TI - Investigating the basis of substrate recognition in the pC221 relaxosome. AB - The nicking of the origin of transfer (oriT) is an essential initial step in the conjugative mobilization of plasmid DNA. In the case of staphylococcal plasmid pC221, nicking by the plasmid-specific MobA relaxase is facilitated by the DNA binding accessory protein MobC; however, the role of MobC in this process is currently unknown. In this study, the site of MobC binding was determined by DNase I footprinting. MobC interacts with oriT DNA at two directly repeated 9 bp sequences, mcb1 and mcb2, upstream of the oriT nic site, and additionally at a third, degenerate repeat within the mobC gene, mcb3. The binding activity of the conserved sequences was confirmed indirectly by competitive electrophoretic mobility shift assays and directly by Surface Plasmon Resonance studies. Mutation at mcb2 abolished detectable nicking activity, suggesting that binding of this site by MobC is a prerequisite for nicking by MobA. Sequential site-directed mutagenesis of each binding site in pC221 has demonstrated that all three are required for mobilization. The MobA relaxase, while unable to bind to oriT DNA alone, was found to associate with a MobC-oriT complex and alter the MobC binding profile in a region between mcb2 and the nic site. Mutagenesis of oriT in this region defines a 7 bp sequence, sra, which was essential for nicking by MobA. Exchange of four divergent bases between the sra of pC221 and the related plasmid pC223 was sufficient to swap their substrate identity in a MobA-specific nicking assay. Based on these observations we propose a model of layered specificity in the assembly of pC221-family relaxosomes, whereby a common MobC:mcb complex presents the oriT substrate, which is then nicked only by the cognate MobA. PMID- 16689806 TI - Greater allergenicity of topical ketoprofen in contact dermatitis confirmed by use. AB - The use of topical non-steroidal anti-inflammatory drugs (NSAIDs) is very popular in spite of their doubtful efficacy and high number of generally not serious, but preventable, adverse effects, especially photoallergy. The allergenic potential of different topical NSAIDs was determined by performing a retrospective observational study of the period 1996-2001 and comparing the cases of allergy and photoallergy with the use of each topical NSAID. The diagnoses were obtained from a review of the clinical records of patch/photopatch testing carried out in the dermatology departments of 2 public hospitals in Bizkaia (Spain). The use of the different topical NSAIDs was obtained from invoices sent to the National Health System and the Reporting odds ratio (ROR) and Proportional reporting ratio (PRR) disproportionality estimates of the FEDRA database of the Spanish Pharmacovigilance System. A total of 139 contact reactions to topical NSAIDs were found with ketoprofen being responsible for 28% of the allergies and 82% of the contact photoallergies in spite of not being the most used topical NSAID (third in the ranking, diclofenac was the first). The ROR for ketoprofen was 3.9 (2.4 6.4) and the PRR 3.4 (2.1-5.5), thus confirming the possibility of a warning signal. The results support the need for regulatory action on topical ketoprofen. PMID- 16689807 TI - Determination of hexavalent chromium in cosmetic products by ion chromatography and postcolumn derivatization. AB - Chromium hydroxide green [Cr(2)O(OH)(4)] and chromium oxide green (Cr(2)O(3)) are colouring agents for use in cosmetic products. These colourants may contain chromium (VI), which cause skin allergies through percutaneous adsorption on the skin. Eye shadow is a representative cosmetic product in which significant colourants are used. We analysed the chromium (VI) in the eye shadows by ion chromatography and post column derivatization. We optimize conditions of chromium (VI) analysis in eye shadows. During the pretreatment procedure, there are no exchange of chromium (III) to chromium (VI). This method has a limit of quantification for chromium (VI) of 1.0 microg l(-1), recovery rate of 100 +/- 3% and analysis time less than 10 min. This result is 300 times more sensitive than the high-performance liquid chromatography method. We applied the optimized method to analyse 22 eye shadows and 6 colouring agents. 2 out of 22 of the products contained more than 5 mg l(-1). In our previous work, 5 mg l(-1) of Cr represented a threshold level. There was much more Cr(VI) in the colouring agents. The Cr(VI) in one of the colouring agents was 97.6 mg l(-1). PMID- 16689808 TI - Skin capacitance imaging and corneosurfametry. A comparative assessment of the impact of surfactants on stratum corneum. AB - Silicon image sensor (SIS) technology was recently introduced as an innovative tool (SkinChip, L'Oreal) providing sensitive imaging of the skin capacitance. This method can detect discrete focal variations in skin surface hydration, and thus early discrete manifestations of skin irritation induced by surfactants. In the present in vivo study, 2 neat and diluted shampoos, and 5% and 10% sodium laurylsulfate solutions were tested on human skin. Each surfactant solution was gently rubbed on the skin using wet hair wicks mimicking the casual use of a shampoo on the scalp. Clinical and SIS evaluations were carried out. In addition, the same products were tested using the ex vivo corneosurfametry bioassay performed on human stratum corneum (SC) harvested by cyanoacrylate skin surface strippings. The colourimetric index of mildness (CIM) was measured on these samples. The product reactivity with the SC was recognized by darker skin capacitance images, and by both lowered SkinChip-generated values and lowered CIM values. The extent in changes varied according to the nature of the test products and their concentrations. The SkinChip image changes likely corresponded to the acute surfactant-induced water swelling of the corneocytes. Skin capacitance imaging and corneosurfametry allow to disclose discrete surfactant-induced alterations of corneocytes. PMID- 16689809 TI - Educational and dermatological aspects of secondary individual prevention in healthcare workers. AB - Healthcare workers (HCW) have an increased risk of suffering from occupational skin diseases (OSDs). Therefore, we established special prevention and skin protection courses for HCW. Medical aims are to optimize diagnostic procedures and individual therapy, educational aims are to improve individual skin protection/skin care habits of the patient and also to influence the participants' attitudes towards health. Participants are patients that have been suspected to suffer from an OSD and are insured with the German Accident Prevention Insurance Association for Health Care Workers (BGW). Teaching units mainly focus on the texture and functioning of the skin, general aspects of OSDs, general information concerning skin protection, instructions about the correct use of skin cleansing and skin protection products. Besides, every participant is clinically examined by a dermatologist and a precise patient's history is obtained. Individually adapted skin protection strategies are developed. Patients can ask the dermatologist questions in confidence. 504 patients (mean age: 36.9 years, SD = 11.7) participated in the skin protection courses. 94.6% (n = 477) suffered from hand eczema frequently caused by a mixture of atopic, irritant and allergic contact dermatitis, but irritant contact dermatitis was the most frequent diagnosis (55.4%, n = 279). The participants rated the course as good to excellent. Health education and advisory services in occupational dermatology are still fragmentary. Prevention of OSD and maintenance of health through education are important complementary measures for dermatological care. In the future, similar educational programmes should be offered for employees of other professions with an increased risk for OSD. PMID- 16689810 TI - Skin condition associated with intensive use of alcoholic gels for hand disinfection: a combination of biophysical and sensorial data. AB - Although hand hygiene is an important and inexpensive measure to prevent nosocomial infections in clinical settings, the compliance of healthcare workers remains low. In Europe, alcoholic hand disinfection is first choice, but there exists a limited user acceptability due to estimated adverse effects on skin condition. This study was designed to investigate skin tolerance to alcohol-based disinfecting gels and changes in skin condition depending on humectant concentration, alcohol grades, as well as type of alcohol used. A comparison of 6 alcohol-based gels was made based on a randomized double-blind study under in use conditions for 1 day. Skin condition was evaluated by measuring transepidermal water loss (TEWL), stratum corneum hydration, apparent skin pH, redness and degree of scaliness. With respect to user acceptability, all gels were sensorially evaluated using a questionnaire. We saw that none of the alcohol based gels, applied under in use conditions, altered TEWL or caused irritation. All gels hydrated the skin, proportionally to their glycerine content, and decreased skin pH. Elevated ethanol concentrations resulted in increased scaliness. Sensorial assessment revealed less appreciation for isopropanol. From this study, it was concluded that gels containing an elevated glycerine concentration and 70% (v/v) ethanol are preferred. PMID- 16689811 TI - Formaldehyde in reusable protective gloves. AB - Due to the clinical findings in a single patient's case, formaldehyde was suspected to be present in clinically relevant levels in reusable protective gloves. Therefore, 9 types of gloves were investigated with the semi-quantitative chromotropic acid method. It was found that 6/9 gloves emitted some formaldehyde and that 4/9 gloves emitted > or =40 microg of formaldehyde. Most of the formaldehyde was found on the inside of the gloves. To get an indication of the clinical relevance, a comparison with a protective cream declared to contain the formaldehyde-releasing agent diazolidinyl urea was performed by comparing areas of gloves with areas of cream layers with thickness 1-2 mg/cm(2). It was found that the amounts of formaldehyde emitted from the gloves might be in the same range as emitted from a layer of cream. PMID- 16689812 TI - Self-reported skin exposure -- a population-based study. AB - The aim was to study self-reported skin exposure in individuals from the general population with or without hand eczema. In a population-based study in 1996 using postal questionnaires to 3000 individuals, 215 of 2218 (9.7%) reported hand eczema during the past 12 months. 182 (85%) of those with hand eczema and 182 without hand eczema, matched for age and sex, participated in telephone interviews in 1997 regarding exposure to skin irritants at work and in leisure time. No differences were shown in occupational exposure to water, hand washing or chemicals in individuals with or without hand eczema. Women reported more wet exposure than men at work and in leisure time. A correlation was found between occupational wet exposure and wet work at home. Persons in high-risk occupations reported more frequent exposure to skin irritants. However, 53% in high-risk occupations reported exposure to water and detergents for less than half-hour a day and 11% in low-risk occupations for more than half-hour a day. In conclusion, individuals with or without hand eczema seem to have similar exposure to skin irritants. Using job titles as a proxy for exposure gives misclassification, which may result in underestimation of the hand eczema risk. PMID- 16689813 TI - Cr(III) and Cr(VI) in leather and elicitation of eczema. AB - The aim of the present study was to investigate the relation between the content of Cr(VI) and soluble Cr(III) in leather and the ability of the leather to elicit eczema in chromium allergic patients. An array of chromium-tanned leather samples was analysed for the content of total Cr(VI) and soluble Cr(III) using the DIN 53314 and the DS/EN 420 methods. Subsequently, a group of 15 patients with a history of foot eczema and leather exposure was exposed to a selection of 14 chromium- and 1 vegetable-tanned leather sample on the upper back for 48 hr. In addition, one leather sample was used for a prolonged 14-day exposure study. In total, 4 of the 15 patients reacted to at least one leather sample, and 5 of the 14 leather samples elicited a reaction in at least 1 patient. The prolonged exposure study demonstrated that an extended exposure period may reveal allergenic potential of a leather sample not otherwise identified using an ordinary 48-hr exposure period. No relation was observed between the measured content of Cr(VI) and soluble Cr(III) in the leather and the elicitation of eczema. Thus, in order to evaluate the quality of chromium-tanned leather in relation to preventing allergic skin reactions, other more clinical relevant methods reflecting the actual bioavailable Cr(III) and Cr(VI) fractions should be developed. PMID- 16689814 TI - Hydrolysed wheat proteins present in cosmetics can induce immediate hypersensitivities. AB - Cosmetics containing hydrolysed wheat proteins (HWP) can induce rare but severe allergic reactions. 9 patients, all females without common wheat allergy, but with contact urticaria to such cosmetics, were studied. 6 of them also experienced generalized urticaria or anaphylaxis to foods containing HWP. All patients had low to moderate levels of immunoglobulin (Ig)E specific of wheat flour (f4) or gluten (f79). Their sensitivity to HWP and their tolerance to unmodified wheat proteins extracted from grains were confirmed using skin tests. Immunoblotting analyses showed that IgE from all patients reacted with almost all HWP tested. Reactions generally occurred with large random peptide aggregates. IgE reacted also with unmodified grain proteins, which contrasted with skin tests results. They reacted always with salt soluble proteins but variably with gluten proteins. No reaction occurred with gliadins in patients without associated immediate hypersensitivity to food containing HWP. These results show the role of hydrolysis on the allergenicity of wheat proteins, both through skin or digestive routes. At least part of the epitopes involved is pre-existing in unmodified wheat proteins. The aggregation of peptide bearing these epitopes and others created by hydrolysis, along with the increased solubility and the route of exposure, are possible factors of the allergenicity of HWP. PMID- 16689815 TI - Pitfalls in drug allergy skin testing: false-positive reactions due to (hidden) additives. AB - Skin testing for drug hypersensitivity is a valuable diagnostic tool and a well established procedure for betalactams and corticosteroids. Although appropriate patch, prick or intracutaneous testing usually permits unequivocal interpretation, the possibility of false-positive reactions must be kept in mind. We report two patients, one with a suspected penicillin allergy and one with a suspected steroid allergy, in whom positive skin tests initially yielded a false diagnosis. In both cases, other findings were inconsistent with the positive skin tests. Therefore, further investigations were performed. After the test preparations had been analysed, the skin test was extended to include the additives, which then yielded the diagnosis of sensitization to the surfactant polysorbate 80 (PS 80, contained in the penicillin test solution) and the emulsion stabilizer carboxymethylcellulose (CMC, contained in the steroid preparation), respectively. It should be noted that positive reactions to test solution additives, as in the first case, may lead to a lifelong false diagnosis of (penicillin) allergy, whereas an undiagnosed allergy to additives in parenteral applications, as in the second case, might prove fatal. PMID- 16689816 TI - Octocrylene: really non-allergenic? PMID- 16689817 TI - Allergic contact cheilitis. PMID- 16689818 TI - Mechanical irritation triggering allergic contact dermatitis from essential oils in a masseur. PMID- 16689819 TI - Contact allergy to triphenyl phosphate: probable cross-reactivity to triphenyl phosphite present in an EN46001 System 22 clear oxygen facemask. PMID- 16689820 TI - Allergic contact dermatitis to petrolatum. PMID- 16689821 TI - Mechanical onycholysis of 2 fingernails in a 7-year-old girl. PMID- 16689822 TI - Neural tube defects and associated anomalies in a fetal and perinatal autopsy series. AB - Neural tube defects (NTDs) are congenital malformations of the central nervous system (CNS) secondary to abnormal closure of the neural tube during embryonic development. This study provides information on NTD rate, distribution, associated morphologic anomalies and organ weights in a Danish fetal and perinatal autopsy series during a 16 year period. The data were extracted from the autopsy reports of a consecutive series of 1984 fetal and perinatal autopsies from the Copenhagen area performed in the period 1989-2004. Registered parameters included: The location and morphology of the NTD, associated morphological anomalies, and organ weights. Organ weights were evaluated according to new fetal autopsy standards and grouped as low, normal or high. Ninety-seven NTD cases were found (4.9%): Spina bifida (38 cases), cephalocele (17 cases) and anencephaly (42 cases). 63% of NTD cases had associated morphologic anomalies. Among the most common were hydrocephalus, NTD in another region, and anomalies in the urogenital system. 58% of the NTD cases had abnormal weight of one or more organs. Most notable was low adrenal weight not only in anencephalic fetuses but also in cases with cephalocele, suggesting a possible association. PMID- 16689823 TI - Post-transplant reactivation of hepatitis C virus: lymphocyte infiltration and the expression of adhesion molecules and their ligands in liver allografts. AB - Hepatitis C virus (HCV) recurrence after liver transplantation has been associated with chronic rejection. Biopsies from 10 patients with post-transplant HCV were examined for expression of adhesion molecules ICAM-1, VCAM-1, and ELAM 1, number of lymphocytes positive for their ligands LFA-1, VLA-4, and SLeX, and activation markers MHC class II antigens and IL2-R by immunohistochemistry. The phenotypes of the graft-infiltrating lymphocytes were determined. Results were compared to those for patients with normal graft function or rejection. Five recipients with HCV reactivation and one with de novo HCV had a biopsy available showing induction of ICAM-1 in sinusoidal endothelium (p<0.05) and hepatocytes (p<0.01), and Class II antigens in hepatocytes (p<0.01), compared to normal controls. Lymphocytes in the graft infiltrate expressed LFA-1, VLA-4, and Class II antigens, but IL2-R was not significantly expressed. CD3+, CD4+, and CD8+ cells were observed. In our study, HCV recurrence was not associated with acute or chronic rejection, and the inflammation was due to the viral infection. PMID- 16689824 TI - Lawsonia intracellularis infection in the large intestines of pigs. AB - In this study we examined the proliferative enteropathy, caused by the obligate intracellular bacterium Lawsonia intracellularis, in colon of naturally infected pigs, using immunohistochemistry, in situ hybridisation and scanning confocal laser microscopy. When 396 pigs submitted for routine laboratory examination were investigated, large intestinal gross lesions were seen in 93, including 74 cases of L. intracellularis colitis (proliferative enteropathy). Fifty-one pigs without recorded colonic gross lesions revealed L. intracellularis colitis microscopically. In four cases, L. intracellularis was only revealed in colon. Fifty-seven pigs were positive for L. intracellularis in the small intestines only. Thus, the overall prevalence of colonic infection in L. intracellularis positive animals was as high as 69% (125 out of 182). In comparison, the large intestinal pathogens Brachyspira hyodysenteriae and Salmonella enterica were only isolated from 5 and 4 of the 93 cases, respectively. Morphologically, an unforeseen severe involvement of the subepithelial mucosa with multiple L. intracellularis found free and within large macrophages was observed in areas with acute infection. The distribution of whole L. intracellularis organisms was confirmed by in situ hybridisation and scanning confocal laser microscopy. The significance and possible role of subepithelial infection in the proliferative enteropathy is discussed. In conclusion, the study shows that L. intracellularis is a prevalent cause of naturally acquired colitis in pigs. PMID- 16689825 TI - Anti-lipopolysaccharide antibodies in acute appendicitis detected by ELISA. AB - In acute appendicitis the bowel transmissibility of the intestinal flora increases in relation to inflammation and edema formation. We can therefore observe an immunologic response in patients, which is detectable using different bacteria isolated from the normal intestinal flora. Our aim was to measure this immunologic reaction and to detect the relationship between this response and histologic types of acute appendicitis. Sera from 47 cases, comprising 38 patients suffering from appendicitis and 9 healthy controls, were examined. The sera were taken shortly before appendectomy and 14 days after operation. The antigens were lipopolysaccharides (LPS) extracted from bacteria of normal intestinal flora: Escherichia coli O21, O22, O33, O61, O68, Bacteroides fragilis and an absolute rough mutant: Shigella sonnei Re 4350. Antibodies were detected by ELISA. We showed a direct relationship between severity of appendicitis and registered antibody titer. Both aerobic and anaerobic bacteria play a role in infection in appendicitis. According to our serologic results the synergy of B. fragilis with E. coli from normal flora is more important in the initiation of inflammation, but in the perforation process the role of E. coli seems more important compared to that of B. fragilis. PMID- 16689826 TI - Altered expression of goblet cell- and mucin glycosylation-related genes in the intestinal epithelium during infection with the nematode Nippostrongylus brasiliensis in rat. AB - Intestinal nematode infection induces marked goblet cell hyperplasia and mucus secretion, but the mechanisms of regulation of the changes still remain to be elucidated. In the present study, epithelial cells were isolated from the rat small intestine at various times after Nippostrongylus brasiliensis infection, and the levels of expression of goblet cell- and mucin glycosylation-related genes were estimated by semi-quantitative reverse transcription (RT)-PCR. Among the genes investigated, mucin core peptide (MUC) 2, sialyltransferase (Siat) 4c and trefoil factor family (TFF) 3 were upregulated as early as 2-4 days post infection, suggesting that they are associated with an early innate protective response. Seven days post-infection and thereafter, when the nematodes reached maturity, significant upregulation of MUC3, MUC4, resistin-like molecule beta (Relmbeta) and 3O-sulfotransferase (3ST)1 was observed, while 3ST2 expression levels increased after the majority of the worms were expelled from the intestine. Similar alterations of glycosylation-related gene expression were also observed in mast-cell-deficient Ws/Ws rats, suggesting that mast cells in the epithelium are not relevant to the upregulation of these genes. The present finding that the expression level of each goblet cell- or glycosylation-related gene was altered differently during the time course of infection indicates the progression of sequential qualitative changes in the mucus layer after infection. PMID- 16689827 TI - Detection of Chlamydophila pneumoniae IgG in paired serum samples: comparison of serological techniques in pneumonia cases. AB - Serological diagnosis of Chamydophila pneumoniae is usually undertaken by complement fixation test (CFT) or by microimmunofluorescence (MIF). A number of commercial methods for detecting C. pneumoniae-specific IgG have been developed. The aim of this study was to compare the performance characteristics of six methods for the diagnosis of pneumonia due to C. pneumoniae, including CFT (in house), MIF (Vircell, Spain), and four ELISAs (Medac, Germany; Savyon, Israel; Serion, Germany; and DRG, Germany). ELISA-Medac, ELISA-Savyon, ELISA-DRG and MIF use C. pneumoniae antigens while ELISA-Serion and CFT use Chlamydophila genus specific antigen. Acute and convalescent samples from 85 pneumonia patients were studied. Using CFT, cases were initially classified as due to Chlamydophila (43 cases); to other agents (23 cases) (influenza A and B, Mycoplasma pneumoniae, respiratory syncytial virus, adenovirus and Legionella pneumophila); or as negative (19 cases). Cases were considered positive if they showed seroconversion, a significant rise in titer or high titer; and were finally classified as positive if they gave a positive result in at least three assays. Sensitivity values ranged from 87% to 97.8%; and specificity from 84.6% to 97.4%. In conclusion, the assays compared appear to be useful tools for the diagnosis of pneumonia due to Chlamydophila. PMID- 16689828 TI - Otomicroscopic findings and systemic interleukin-6 levels in relation to etiologic agent during experimental acute otitis media. AB - The aim of the present study was to explore whether it was possible to differentiate the clinical course and the otomicroscopic appearance of acute otitis media (AOM) caused by common otitis pathogens in an animal model. Systemic interleukin (IL)-6 levels as early markers for bacterial AOM were also studied. Four groups of rats were inoculated with either Streptococcus pneumoniae, Streptococcus pyogenes, non-typeable Haemophilus influenzae or Moraxella catarrhalis. The animals were monitored by otomicroscopy, photos of the tympanic membrane, cultures and IL-6 detection in serum the following 4 days. The gram positive S. pneumoniae and S. pyogenes induced severe AOM with opaque effusion behind the tympanic membrane, pronounced dilation of the vessels and spontaneous perforations. The gram-negative H. influenzae and M. catarrhalis induced a less severe infection with cloudy, sometimes foamy effusion, and no spontaneous perforations. With the otomicroscopic findings it was possible to distinguish between infections induced by gram-positive bacteria and gram-negative bacteria. Detection of interleukin-6 in serum appeared to be of limited use for all infections except the pneumococcal AOM, but this needs to be further investigated. PMID- 16689829 TI - Mutational analysis of proapoptotic caspase-9 gene in common human carcinomas. AB - Mounting evidence indicates that deregulation of apoptosis is involved in the mechanisms of cancer development. Caspase-9 plays a crucial role in the initiation phase of the intrinsic apoptosis pathway. To explore the possibility that the genetic alterations of caspase-9 might be involved in the development of human cancers, we analyzed the entire coding region and all splice sites of the human caspase-9 gene for the detection of somatic mutations in a series of 353 cancers, including 180 gastric, 104 colorectal and 69 lung adenocarcinomas. Overall, we detected three somatic mutations of caspase-9, but all of the mutations were silent mutations. The mutations were observed in 2 of 104 colorectal carcinomas and 1 of 180 gastric carcinomas. These data indicate that the caspase-9 gene is rarely mutated in gastric, colorectal and lung adenocarcinomas, and suggest that caspase-9 gene mutation may not contribute to the pathogenesis of these cancers. PMID- 16689830 TI - Interspecies variation in Candida biofilm formation studied using the Calgary biofilm device. AB - An in vitro assay to study multiple Candida biofilms, in parallel, has been carried out using the Calgary biofilm device (CBD). We here report: i) standardization of the CBD for Candida albicans biofilm formation, ii) kinetics of C. albicans biofilm formation, iii) biofilm formation by five Candida species, and iv) effect of dietary carbohydrates on biofilm formation. The biofilm metabolic activity on all CBD pegs was similar (p=0.6693) and C. albicans biofilm formation revealed slow growth up to 36 h and significantly higher growth up to 48 h (p<0.001). Significant differences in total biofilm metabolic activity were seen for glucose, fructose and lactose grown C. albicans compared with sucrose and maltose grown yeasts. Candida krusei developed the largest biofilm mass (p<0.05) relative to C. albicans, C. glabrata, C. dubliniensis and C. tropicalis. Scanning electron microscopy revealed that C. krusei produced a thick multilayered biofilm of pseudohyphal forms embedded within the polymer matrix, whereas C. albicans, C. dubliniensis and C. tropicalis biofilms consisted of clusters or chains of cells with sparse extracellular matrix material. We conclude that CBD is a useful, simple, low cost miniature device for parallel study of Candida biofilms and factors modulating this phenomenon. PMID- 16689831 TI - Urokinase plasminogen activator receptor (uPAR) expression is reduced by tyrosine kinase inhibitors. AB - Previously we reported that tyrosine kinase inhibitors (TKI) produced a reduction in uPA expression in prostatic cancer cells, and that TKI-treated cells were less invasive compared to untreated cells. Nevertheless, no change in cell migration was observed when TKI-treated cells were supplied with external uPA, thus indicating more complex mechanisms leading to decreased cell invasion. uPAR expression was measured with an enzyme-linked immunosorbent assay (ELISA) in PC-3 and DU-145 prostate carcinoma cells treated with the two TKI genistein and AG 1478. uPAR mRNA levels were measured with real-time reverse transcriptase polymerase chain reaction (RT-PCR). uPAR immunocytochemistry was used to examine the receptor distribution in cells grown on a reconstituted basal lamina. Immunocytochemistry showed an intense uPAR immunostaining in invading cells, particularly in the leading edge membrane. Treatment with genistein and AG-1478 led to a decreased expression of uPAR in DU-145, but not in PC-3. Furthermore, a reduction of uPAR mRNA was found in TKI-treated DU-145 cells, while PC-3 was not affected. Our results indicate a possible role of TKI as cancer suppressors by acting as a regulator of uPAR expression. PMID- 16689832 TI - Female urethral adenocarcinoma with a heterogeneous phenotype. AB - We here report a very rare case of female urethral adenocarcinoma. A 77-year-old woman presented with urinary retention. Cystoscopy showed a urethral tumor and the biopsy material showed adenocarcinoma. Macroscopically, the tumor measuring 3.0 x 3.0 x 2.4 cm was predominantly observed around the periurethral area on the proximal side. Histologically, patterns of columnar/mucinous adenocarcinoma, clear cell adenocarcinoma and papillary/micropapillary carcinoma were observed, but there was no evidence of a cribriform pattern. Immunohistochemically, neoplastic cells of at least one of three components were positive for CK7 and CK20 or CA125. We suggest that female urethral adenocarcinoma with a histologically and immunohistochemically heterogeneous phenotype may originate from cells within urethral or paraurethral tissue, such as urethritis glandularis or intestinal metaplastic epithelium and Mullerian tissue. PMID- 16689833 TI - In memoriam: special tribute to Seymour Furman, M.D. 1931-2006. PMID- 16689834 TI - Seymour Furman. PMID- 16689835 TI - Seymour Furman. PMID- 16689836 TI - Seymour Furman. PMID- 16689837 TI - An intracardiac pacemaker for Stokes-Adams seizures. PMID- 16689838 TI - The dark side of the new era. PMID- 16689839 TI - Effectiveness of excimer laser-assisted pacing and ICD lead extraction in children and young adults. AB - BACKGROUND: High capture thresholds, decreased electrical sensing, and lead fractures continue to be a problem in children undergoing transvenous pacing. The clinician must therefore decide at the time of pacing system revision to either abandon chronically implanted transvenous pacing leads or extract them. METHODS: We report our experience using an excimer laser-assisted (LA) strategy for removing chronically implanted pacing (36) and implantable cardioverter/defibrillator (ICD) (7) leads in children and young adults. The study population consisted of 25 patients, in whom 29 procedures were performed. The patients ranged in age from 8.4 to 39.9 years, median age was 13.9 years, at the time of the extraction procedure. In all procedures, a Spectranectics locking stylet and excimer laser sheath were used to assist in lead extraction. RESULTS: Lead removal was complete for 39 (91%) leads, and partial for four leads. In two patients, the pacing lead tip was retained and in two, the ring electrode from a bipolar pacing lead was left in situ. All ICD leads were removed completely. Two major complications occurred--cardiac perforation and tamponade (1), and thrombosis of the left subclavian/innominate vein (1). LA extraction facilitated the implantation of new pacing or ICD leads in three patients with obstructed venous access. CONCLUSIONS: Removal of pacing and ICD leads using an excimer LA technique was highly successful. Lead removal was complete in 91%. The most common indication for lead removal in our study was lead fracture. Complications were few, but may be significant. PMID- 16689840 TI - Automatic implantable cardioverter-defibrillators in Chagas' heart disease patients with malignant ventricular arrhythmias. AB - A total of 46 consecutive Chagas' disease patients had an automatic cardioverter defibrillator implanted at our institution from October 1998 to January 2004. A retrospective longitudinal study was carried out to identity type of life threatening ventricular arrhythmias as well as type of therapy delivered. Of these, 41 (91%) had been recovered from cardiac arrest. Five (15%) of 33 patients in whom echocardiography was done had no left ventricular function. Antiarrhythmic therapy was delivered to 37 (80%) patients during postimplant follow-up. Thirty-one of 37 (84%) patients received both shock and antitachycardia pacing, five (13%) only antitachycardia pacing, and one (3%) patient only shock. Median time to first shock was 16 days, varying from 1 to 576 days. Ventricular fibrillation was the cause of first shock in 12 patients (32%), ventricular tachycardia in 11 (29%), and ventricular tachycardia not responding to antitachycardia pacing degenerating into ventricular fibrillation in nine (24%). Five patients with ventricular tachycardia were treated with antitachycardia pacing. Probability of freedom from device discharged was 47% at 90 days, 34% at 180 days, and 9% at 360 days in the postimplant follow-up. Thus, patients with chronic Chagas' heart disease recovered from cardiac arrest have a peculiar arrhythmogenic profile characterized by a high frequency of ventricular fibrillation and no left ventricular systolic dysfunction and a short period of time for first shock. PMID- 16689841 TI - Heart rate variability following neonatal heart surgery for complex congenital heart disease. AB - BACKGROUND: Altered cardiac autonomic control may play a role in the morbidity and mortality suffered by neonates who undergo surgery for complex congenital heart disease (CHD). The purpose of this study was to evaluate cardiac autonomic activity, as measured by spectral indices of heart rate variability (HRV), prior to and early after infant surgery for CHD and attempt to correlate HRV indices with clinical outcome. In addition, we assessed the hypothesis that single ventricle physiology and surgical interruption of the great arteries negatively affects HRV. METHODS: Sixty neonates prospectively wore 24-hour Holter monitors at three time points: before and early after CHD surgery, and at 3- to 6-month follow-up. Standard spectral indices of HRV were measured. RESULTS: In the early postoperative time point, patients with single-ventricle physiology had lower low frequency power (LF) compared to patients with two ventricles (P=0.040). Surgical interruption of the great arteries did not affect HRV in this cohort. For the entire cohort, LF (P=0.004) and high-frequency power (HF) (P<0.001) increased over the three time points, while LF/HF (P=0.119) did not significantly change. In the multivariable linear regression model, significant predictors of longer postoperative hospital stay included longer total support time (P=or<0.001), longer duration of inotrope support (P=0.012), elevated mean heart rate at postoperative time point (P=0.002), and lower LF/HF ratio at the postoperative time point (P=0.014). CONCLUSION: Patients with single-ventricle physiology have a significant physiologic reduction in LF in the early postoperative period compared to patients with two ventricles. Diminished cardiac autonomic control is associated with longer hospitalization following neonatal cardiac surgery. PMID- 16689842 TI - Circadian variation of beat-to-beat QT interval variability in patients with prior myocardial infarction and the effect of beta-blocker therapy. AB - BACKGROUND: Recent studies have demonstrated that increased QT interval variability (QTV) is associated with a greater susceptibility to ventricular arrhythmias and that patients with prior myocardial infarction (MI) were prone to ventricular arrhythmias during the daytime. The goal of the present study was to investigate the circadian variation of the QTV and to determine whether beta blocker therapy improves the temporal fluctuation of the ventricular repolarization in patients with MI. METHODS: The study population consisted of 15 MI patients who had not received beta-blocker therapy, 11 MI patients who had received beta-blocker therapy, and 12 healthy subjects. Twenty-four hour Holter monitoring was obtained, and the RR and QT intervals were calculated automatically from 512 consecutive sinus beats for every 2 hours. RESULTS: In the daytime, the QT-SD was significantly greater in the MI group than in the healthy subjects (P<0.01), but there was no difference in the QT-SD when comparing the beta-blocker group to the control group. Moreover, the QT variability index and the QT variance normalized for the mean QT were similar pattern with QT-SD. The heart rate variability did not significantly differ when compared between the three study groups. CONCLUSION: These data indicate that the QTV increases during the daytime in patients with MI and that this circadian effect is prevented by beta-blocker therapy. Thus, beta-blocker therapy may reverse the maladaptation of the ventricular repolarization to the change in the heart rate and may thereby reduce the ventricular arrhythmias and decrease the mortality in patients with MI. PMID- 16689843 TI - High success rate with cryomapping and cryoablation of atrioventricular nodal reentrytachycardia. AB - INTRODUCTION: Cryoablation is a new alternative to radiofrequency (RF) ablation for treatment of atrioventricular nodal reentry tachycardias (AVNRT). Mapping with reversible effect on the arrhythmia substrate or the AV node can be done before irreversible ablation is performed. This study evaluates an approach with systematic cryomapping, ablating only in areas with prompt effect on the arrhythmia substrate and evaluates whether the success rates and procedure times are similar to RF ablation. METHODS AND RESULTS: Seventy-five consecutive patients with typical slow-fast AVNRT were studied. Cryomapping at -30 degrees C was performed before ablation with a goal temperature of -70 degrees C for 240 seconds. The ablation procedure was successful in 74 of the 75 patients, giving an acute success rate of 99%. During a mean follow-up of 338 days, 70 of the 74 primarily successfully ablated patients were free from the treated arrhythmia, giving a recurrence rate of 5% and a total success rate of 93%. Total procedure time including a 30-minute test after successful ablation was 126+/-55 minutes. Fluoroscopy time was 18.5+/-14.9 minutes. CONCLUSION: Cryoablation of AVNRT appears to be as effective as RF ablation both acute and in long term with minimal risks for unwanted injuries on the conduction system. The procedure can be done with reasonable procedure and fluoroscopy times. PMID- 16689844 TI - Electrophysiologic characteristics and radiofrequency catheter ablation in children with Wolff-Parkinson-White syndrome. AB - BACKGROUND: The majority of cardiac arrhythmias in children are supraventricular tachycardia, which is mainly related to an accessory pathway (AP)-mediated reentry mechanism. The investigation for Wolff-Parkinson-White (WPW) syndrome in adults is numerous, but there is only limited information for children. This study was designed to evaluate the specific electrophysiologic characteristics and the outcome of radiofrequency (RF) catheter ablation in children with WPW syndrome. METHODS: From December 1989 to August 2005, a total of 142 children and 1,219 adults with atrioventricular reentrant tachycardia (AVRT) who underwent ablation at our institution were included. We compared the clinical and electrophysiologic characteristics between children and adults with WPW syndrome. RESULTS: The incidence of intermittent WPW syndrome was higher in children (7% vs 3%, P=0.025). There was a higher occurrence of rapid atrial pacing needed to induce tachycardia in children (67% vs 53%, P=0.02). However, atrial fibrillation (AF) occurred more commonly in adult patients (28% vs 16%, P=0.003). The pediatric patients had a higher incidence of multiple pathways (5% vs 1%, P<0.001). Both the onset and duration of symptoms were significantly shorter in the pediatric patients. The antegrade 1:1 AP conduction pacing cycle length (CL) and antegrade AP effective refractory period (ERP) in children were much shorter than those in adults with manifest WPW syndrome. Furthermore, the retrograde 1:1 AP conduction pacing CL and retrograde AP ERP in children were also shorter than those in adults. The antegrade 1:1 atrioventricular (AV) node conduction pacing CL, AV nodal ERP, and the CL of the tachycardia were all shorter in the pediatric patients. CONCLUSION: This study demonstrated the difference in the electrophysiologic characteristics of APs and the AV node between pediatric and adult patients. RF catheter ablation was a safe and effective method to manage children with WPW syndrome. PMID- 16689845 TI - The effect of the fiber curvature gradient on break excitation in cardiac tissue. AB - BACKGROUND: Break excitation has been hypothesized as a mechanism for the initiation of reentry in cardiac tissue. One way break excitation can occur is by virtual electrodes formed due to a curving fiber geometry. In this article, we are concerned with the relationship between the peak gradient of fiber curvature and the threshold for break stimulation and the initiation of reentry. METHODS: We calculate the maximum gradient of fiber curvature for different scales of fiber geometry in a constant tissue size (20x20 mm), and also examine the mechanisms by which reentry initiation fails. RESULTS: For small peak gradients, reentry fails because break excitation does not occur. For larger peak gradients, reentry fails because break excitation fails to develop into full-scale reentry. For strong stimuli above the upper limit of vulnerability, reentry fails because the break excitation propagates through the hyperpolarized region and then encounters refractory tissue, causing the wave front to die. PMID- 16689846 TI - Endocardial visualization of esophageal-left atrial anatomic relationship by three-dimensional multidetector computed tomography "navigator imaging". AB - BACKGROUND: The close proximity of left atrium (LA) and esophagus during radiofrequency ablation for atrial fibrillation (AF) predisposes to thermal injury resulting in atrio-esophageal fistula. This work proposes to study the anatomic relationship between the esophagus and the LA wall using multidetector computed tomography (MDCT) three-dimensional (3D) "Navigator" reconstruction technique. METHODS: Forty-five consecutive patients (37 men, mean age 52.7+/-14.1 years) with preradiofrequency ablation MDCT scans of the thorax for AF were recruited. Length and type (continuous or interrupted) of fat pad between esophagus and LA were evaluated. The position, width, and length of the esophagus in contact (without fat pad) with the LA were determined by using "Navigator" software on the endocardial view of LA. RESULTS: The fat pad was continuous in 4% (2 of 45) and interrupted in 96% (43 of 45) patients. The mean width and length of esophageal-LA contact in 43 cases with interrupted fat pad was 24.0+/-5.8 mm (range 10.5-35.3 mm) and 41.9+/-11.6 mm (5.4-64 mm), respectively. There was an inverse relationship between the lengths of the esophageal-LA contact and the upper fat pad (r=-0.50, P=0.001). The esophagus was located to the left, right, and midline of the LA in 40, 2, and 1 patients, respectively, and the esophagus was in contact with and overrode the PV orifice in 22 and 4 patients, respectively. CONCLUSION: Direct esophageal-LA contact without the intervening fat pad was present in 96% of the cases, with 93% of esophagi lying to the left of the LA and 51% in contact with a PV orifice. Three-dimensional Navigator imaging technique has enhanced the visualization of the anatomical information of the esophagus, LA wall, and PV orifices that may be used to avoid thermal injury to the esophagus during LA ablation procedure. PMID- 16689847 TI - Computed tomography to assess possible cardiac lead perforation. AB - BACKGROUND: Chest radiography and echocardiography are standard techniques to evaluate possible extracardiac migration of pacemaker and defibrillator leads, but computed tomography (CT) scanning may be a helpful adjunct. METHODS: Chest CT using a 64-detector helical scanner was performed in three cases of lead perforation where standard techniques did not yield a definitive diagnosis. In two patients, a cardiac CT protocol was followed, the primary indication for which was to assess the lead tip location. On one patient, persistent chest pain several days after the implant led to a chest CT to rule out a pulmonary embolism, and the extracardiac lead was found fortuitously. RESULTS: In all cases, CT scan images resulted in a definitive diagnosis of extracardiac migration of pacemaker or defibrillator leads. One patient had an extracardiac atrial lead which was dysfunctional but not associated with symptoms. The risks of repositioning were felt to outweigh potential benefits in this patient with chronic obstructive pulmonary disease, and treatment was conservative. The other two patients had extracardiac ventricular leads and were symptomatic with pleuritic chest pain. These patients were treated by lead revision with no evidence of bleeding into the pericardial space. All patients did well and none have suffered any further complications. CONCLUSION: CT images aid in the diagnosis of lead perforation when other modalities are nondiagnostic. Recent advances in CT technology have been associated with increased use of this technique for evaluation of chest pain, analysis of which should now include location of intracardiac leads. PMID- 16689848 TI - Indices of electrical and contractile remodeling during atrial fibrillation in man. AB - BACKGROUND: Atrial electrical and contractile remodeling have been demonstrated to coincide during atrial fibrillation (AF) in experimental studies. We explored whether electrical and contractile remodeling correlate in man and explored its clinical implications. METHODS: Forty-nine patients with persistent AF were studied. Electrical remodeling was assessed noninvasively using spectral analysis to estimate the average fibrillatory rate (AFR). Atrial contractility was assessed by transesophageal echocardiography (TEE) measurement of left atrial appendage outflow velocity (LAAOV). RESULTS: The AFR was 403+/-43 fibrillations per minute (fpm) and the LAAOV was 0.27+/-0.14 m/s. A significant correlation was found between AFR and LAAOV (r=-0.47, P=0.001). In patients with a LAAOV>or=0.25 m/s, the AFR was 387+/-48 fpm compared to 419+/-31 fpm among patients with LAAOV<0.25 m/s (P<0.01). CONCLUSIONS: This study demonstrates that indices of electrical and contractile remodeling are strongly correlated in persistent AF in man. The interindividual overlap, however, is too large to allow predictions of LAAOV based on fibrillatory frequency alone. PMID- 16689849 TI - Torsade de pointes: an electrophysiological effect of cardiac resynchronization? PMID- 16689850 TI - Atrial fibrillation ablation: reaching the mainstream. AB - INTRODUCTION AND AIMS: Ablation of atrial fibrillation (AF) has evolved rapidly in the decade since its inception. We aimed to review the results of this evolution as reflected in the published literature. METHODS: Publications through 2005 were reviewed, and data included if there was information on the technique used, and follow-up of at least 6 months. RESULTS: More than 23,000 patients met criteria for inclusion. There has been a steady improvement in reported outcomes (P<0.001). Variations on radiofrequency catheter ablation for pulmonary vein isolation result in apparent elimination ("cure") or improvement of AF in 75%, and surgical techniques are even better. CONCLUSIONS: Catheter ablation of AF is now a mainstream procedure. Continuing technical advances are needed to achieve better results with more uniformity and reduced procedure times. PMID- 16689851 TI - The color of silence. PMID- 16689852 TI - Klebsiella pneumoniae: a rare cause of device-associated endocarditis. AB - Intravascular infections involving implanted pacemakers and defibrillators are being seen with increasing frequency. This report describes a case of intravascular infection of an implanted defibrillator with Klebsiella pneumoniae, an unusual pathogen for pacemaker or defibrillator infection. PMID- 16689853 TI - Pacemaker lead endocarditis caused by Staphylococcus hominis. AB - Infective endocarditis related to pacemaker is a rare but serious condition in permanent venous tracing. A 65-year-old man was admitted to the hospital with high fever and chills. A DDD pacemaker had been implanted via the right subclavian vein because of sick sinus syndrome 6 years earlier. Transesophageal echocardiogram identified an oscillating round hyperechoic mass with a stalk near the tricuspid valve. Blood cultures grew Staphylococcus hominis. The patient was treated with antibiotics and operated on after the acute phase of the illness had subsided. We hereby report a case of lead endocarditis caused by S. hominis in a patient with pacemaker, which has been rarely reported in the English literature. PMID- 16689854 TI - Progressive and persistent atrial inexcitability. AB - Sinus node disease is characterized by the presence of significant sinus bradycardia or prolonged sinus pauses, and is attributed to either primary failure of sinus node automaticity or sino-atrial conduction disturbance. We present two patients with symptomatic bradycardia due to idiopathic global atrial inexcitability. PMID- 16689855 TI - Pilsicainide-induced verapamil sensitive idiopathic left ventricular tachycardia. AB - A 20-year-old man was admitted to our hospital for treatment of verapamil sensitive idiopathic left ventricular tachycardia (ILVT). During the electrophysiologic study (EPS), no sustained ventricular tachycardia (VT) could be induced both at baseline and after infusion of isoproterenol. However, sustained clinical VT could be easily induced with single ventricular extrastimulation following intravenous administration of pilsicainide, a class Ic sodium channel blocker. The arrhythmia was ablated with radiofrequency catheter ablation. PMID- 16689856 TI - Emotion-triggered cardiac asystole-inducing neurocardiogenic syncope. AB - The pathophysiology of neurocardiogenic syncope (NCS) is multifactorial. Recurrent syncopal episodes can result in injury and can provoke substantial anxiety among patients. Although an abundance of descriptions of various forms of syncope have been reported in the literature, few articles to date address a documented case due to emotional stress or sound. This is a report of a 31-year old woman who fainted after being startled by someone sneezing. Review of the episode on her event recorder revealed a transient cardiac asystole of 10 seconds. We discuss the incidence of NCS and the proposed mechanism by which this syncopal event occurred. PMID- 16689857 TI - Glucocorticoid therapy-induced skin atrophy. AB - Glucocorticoids (GCs) are highly effective for the topical treatment of inflammatory skin diseases. Their long-term use, however, is often accompanied by severe and partially irreversible adverse effects, with atrophy being the most prominent limitation. Progress in the understanding of GC-mediated molecular action as well as some advances in technologies to determine the atrophogenic potential of compounds has been made recently. It is likely that the detailed mechanisms of GC-induced skin atrophy will be discovered and in vitro models for the reliable prediction of atrophy will be established in the foreseeable future. This knowledge will not only facilitate safety profiling of established drugs but will also foster further drug discovery by improving compound characterization processes. New insights into GC modes of action will guide optimization strategies aiming at novel GC receptor ligands with improved effect/side effect profile. PMID- 16689858 TI - Subtoxic concentrations of allergenic haptens induce LC migration and maturation in a human organotypic skin explant culture model: a novel method for identifying potential contact allergens. AB - The accelerated migration of Langerhans cells (LCs) out of the epidermis and up regulation of maturation markers, upon treatment with subtoxic concentrations of chemicals, were used as the criteria to determine the potential of allergenic chemicals capable of inducing a hapten-specific delayed-type hypersensitivity reaction. Here we report the findings of a study in which seven chemicals, coded and tested in a blind fashion, were classified as contact allergens or non allergens using the human organotypic skin explant culture (hOSEC) model. All chemicals that were identified as a contact sensitizer on decoding induced a definite decrease in the number of CD1a and HLA-DR-positive epidermal LCs in the epidermis of the skin explants, as determined by both semiquantitative immunohistochemistry and quantitative flow cytometric analysis. A significant increase in the number of CD83(+) cells was accompanied by up-regulation of activation molecules in the epidermis of hOSEC exposed specifically to contact allergens. In contrast, there were only minor alterations in epidermal LC numbers, expression of CD83 and other activation markers by LCs when the biopsies were treated with non-toxic concentrations of non-allergenic irritants and vehicles. The data suggest that an increased epidermal LC migration and maturation accompanied by increased expression of activation markers could be used as end-point determinants to screen allergens in a non-animal alternative hOSEC model. PMID- 16689859 TI - Skin irritants and contact sensitizers induce Langerhans cell migration and maturation at irritant concentration. AB - Skin irritants and contact allergens reduce the number of Langerhans cells (LCs). It has been assumed that this reduction is due their migration to the draining lymph node (LN) for initiating immune sensitization in a host. Skin irritation, however, as opposed to contact allergy is not considered to be an immunological disease. Nevertheless, skin irritants are also known for their adjuvant-like effects on contact allergy, resulting in skin hypersensitivity reactions like toxic dermatitis. The human organotypic skin explant culture (hOSEC) model is used to study the characteristics of chemical-induced migration of CD1a(+) LCs out of the epidermis in relation to irritancy or toxicity. We analysed cells emigrating out of hOSEC for CD1a(+) LCs, CD83(+) mature dendritic cells (DCs) and CCR7(+) LN homing cells. After exposure to a toxic concentration of a non immunogenic irritant, an increase of CD1a(+)CD83(+) LCs was found in the culture medium. A non-toxic concentration of an sensitizer induced an increase of CD1a(+) cells. About 50% of skin emigrating CD1a(+) LCs were CD83(-) (immature) but all were CCR7(+). Skin irritation by both non-allergenic and allergenic compounds induces LC migration and maturation. In contrast, only allergenic compounds induced LC migration with partial maturation at subtoxic concentration. This effectively demonstrates that irritation is physiologically needed stimuli for inducing LC maturation. PMID- 16689860 TI - Epidermal caspase-3 cleavage associated with interferon-gamma-expressing lymphocytes in acute atopic dermatitis lesions. AB - Keratinocyte apoptosis mediated by Fas/Fas ligand molecular interactions and subsequent caspase activation is believed to play an important role in the pathogenesis of atopic dermatitis (AD), in particular for the formation of spongiosis. To estimate epidermal caspase activation in normal and AD skin under in vivo conditions, we analysed caspase-3 cleavage by immunohistology. In normal skin as well as non-lesional AD skin, we detected caspase-3 cleavage in single cells of the basal layer. In contrast, in acute lesional AD skin, we not only obtained evidence for increased expression of cleaved caspase-3 in keratinocytes of the basal layer but also observed caspase-3 cleavage in one or more layers of the spinous cell layer, in particular in spongiotic areas. Short-term topical treatment of the skin lesions with tacrolimus or pimecrolimus abolished the expression of cleaved caspase-3 in the spinous layer. Moreover, epidermal caspase 3 cleavage correlated with the numbers of dermal interferon-gamma (IFN-gamma) expressing CD4+ and CD8+ lymphocytes in skin lesions of AD patients, supporting the view that IFN-gamma is important for the activation of proapoptotic pathways in keratinocytes. This is also confirmed by the observation of increased Fas expression on keratinocytes in acute AD lesions that was markedly reduced following topical calcineurin inhibitor treatment. These data suggest that caspase-3 cleavage in the spinous layer of the epidermis is a pathologic event contributing to spongiosis formation in AD, whereas cleavage of caspase-3 in basal cells might represent a physiologic mechanism within the process of epidermal renewal. PMID- 16689861 TI - In situ profiling and quantification of cytokines released during ultraviolet B induced inflammation by combining dermal microdialysis and protein microarrays. AB - In skin, an evolving inflammatory or immune response is triggered by early release of a cytokine cascade into the extracellular space. Investigation of extracellular cytokine secretion in situ has been limited by low cut-off filtering membranes and sample volume size and the inability to monitor changes in cytokine protein levels in real-time in situ. Here, we combine for the first time the methods of intradermal microdialysis and antibody protein arraying to profile the early cascade of multiple cytokines in a complex inflammatory response exemplified by ultraviolet B (UVB)-induced inflammation. We observed significant differences of the cytokine and growth factor responses after tissue injury by catheter placement and UVB-induced inflammation. UVB irradiation initiates a rapid proinflammatory response followed by a mixed TH1/TH2 response in which ultimately TH2 cytokines IL-4 and IL10 predominated after 24 h. This most likely indicates the termination and self limitation of the inflammatory response. We conclude that the combination of dermal microdialysis and protein microarray offers a powerful tool to analyze in real-time the complex and rapidly changing interstitial protein milieu during cutaneous inflammatory responses. PMID- 16689862 TI - Expression of voltage-gated calcium channel subunit alpha1C in epidermal keratinocytes and effects of agonist and antagonists of the channel on skin barrier homeostasis. AB - Recent studies demonstrated that skin surface electric conditions affect epidermal permeability barrier homeostasis. These results suggest the existence of voltage sensor on the keratinocytes of the epidermis. On the contrary, specific blockers of the voltage-gated calcium channel (VGCC) also affect epidermal barrier homeostasis, but the existence and function of the channel has not been determined. We demonstrated here immunohistochemically the expression of the main subunit of the L-type VGCC, alpha1C, which alone has a calcium channel function, in mouse and human epidermis. Immunostaining, RT-PCR, and Western blotting were carried out to detect the channel protein. Messenger RNA of alpha1C was also detected in mouse epidermis and human keratinocyte culture by RT-PCR. We also evaluated the function of the channel in the cultured human keratinocytes. Previously, we demonstrated that influx of calcium ion into epidermal keratinocytes delayed the barrier recovery after barrier disruption and topical application of calcium channel blocker accelerated the barrier recovery. In this study, topical application of nifedipine and R-(+)-BAY K8644 after tape stripping of hairless mice accelerated the barrier repair rate while application of S-(-) BAY K8644 delayed the barrier recovery. These results suggest that the VGCC exists on epidermal keratinocytes and plays an important role in skin barrier homeostasis. PMID- 16689863 TI - Prevention of psoriasis-like lesions development in fsn/fsn mice by helminth glycans. AB - The helminth glycan LNFPIII is an immunomodulatory molecule, driving CD4(+) Th2 type biasing as well as immune suppression. Psoriasis is an autoimmune disease where the immune mechanisms as well as the antigens responsible for development of immune autoreactivity are still not known. In the absence of defined immunological mechanisms, we asked whether LNFPIII would function as novel therapy for psoriasis. We tested the therapeutic efficacy of LNFPIII using the flaky skin (fsn)/fsn mutant mouse model of psoriasis-like lesion development. We found that treatment of mice with LNFPIII prevented the appearance of psoriatic skin lesions on fsn/fsn mice. Examination of the skin 2 weeks after treatment demonstrated that prevention of skin lesions was associated with maintenance of normal epidermis thickness in LNFPIII-treated mice as compared with a significantly thickened epidermis in control treated and diseased mice. In addition, cells from skin of LNFPIII-treated mice produced lower amounts of interferon-gamma as compared with cells from skin of control treated diseased mice. Examination of macrophages and T cells from peripheral lymph nodes of control and LNFPIII-treated fsn/fsn mice showed that glycan treatment reduced the numbers of Gr1(+)F4/80(+) macrophages and the numbers of CD8(+) T cells, restoring the numbers of these two cell populations as well as the CD4 : CD8 ratio to near normal levels. Overall, the results from this study suggest that the helminth immunomodulatory glycan LNFPIII functions to prevent development of psoriatic-like skin lesions in fsn/fsn mice. PMID- 16689864 TI - Influence of calcium on the proteolytic degradation of the calmodulin-like skin protein (calmodulin-like protein 5) in psoriatic epidermis. AB - The calmodulin-like skin protein (CLSP) or so-called calmodulin-like protein 5, a recently discovered skin-specific calcium-binding protein, is closely related to keratinocyte differentiation. The 16-kDa protein is proteolytically degraded in the upper layers of the stratum corneum (SC) of healthy skin. With the use of specific new monoclonal antibodies to CLSP, we were able to demonstrate that the abnormal elevated levels of CLSP, characteristic of psoriatic epidermis, were probably not due to an overexpression of the protein, but most likely the result of its non-degradation. Further in vitro experiments using recombinant CLSP and in situ data clearly showed that calcium protected and chelator accelerated CLSP degradation. These data indicate that CLSP degradation in the SC of psoriatic skin might be hindered by the abnormally elevated calcium concentration. No degradation of CLSP in psoriatic epidermis keeping its ability to bind protein as transglutaminase 3 may have a physiological role in skin diseases such as psoriasis. PMID- 16689866 TI - Cultivation of methanotrophic bacteria in opposing gradients of methane and oxygen. AB - In sediments, methane-oxidizing bacteria live in opposing gradients of methane and oxygen. In such a gradient system, the fluxes of methane and oxygen are controlled by diffusion and consumption rates, and the rate-limiting substrate is maintained at a minimum concentration at the layer of consumption. Opposing gradients of methane and oxygen were mimicked in a specific cultivation set-up in which growth of methanotrophic bacteria occurred as a sharp band at either c. 5 or 20 mm below the air-exposed end. Two new strains of methanotrophic bacteria were isolated with this system. One isolate, strain LC 1, belonged to the Methylomonas genus (type I methantroph) and contained soluble methane mono oxygenase. Another isolate, strain LC 2, was related to the Methylobacter group (type I methantroph), as determined by 16S rRNA gene and pmoA sequence similarities. However, the partial pmoA sequence was only 86% related to cultured Methylobacter species. This strain accumulated significant amounts of formaldehyde in conventional cultivation with methane and oxygen, which may explain why it is preferentially enriched in a gradient cultivation system. PMID- 16689867 TI - Temperature- and pH-dependent accumulation of heat-shock proteins in the acidophilic green alga Chlamydomonas acidophila. AB - Chlamydomonas acidophila, a unicellular green alga, is a dominant phytoplankton species in acidic water bodies, facing severe environmental conditions such as low pH and high heavy metal concentrations. We examined the pH-, and temperature dependent accumulation of heat-shock proteins in this alga to determine whether heat-shock proteins play a role in adaptation to their environment. Our results show increased heat-shock proteins accumulation at suboptimal pHs, which were not connected with any change in intracellular pH. In comparison to the mesophilic Chlamydomonas reinhardtii, the acidophilic species exhibited significantly higher accumulations of heat-shock proteins under control conditions, indicating an environmental adaptation of increased basal levels of heat-shock proteins. The results suggest that heat-shock proteins might play a role in the adaptation of C. acidophila, and possibly other acidophilic algae, to their extreme environment. PMID- 16689868 TI - Microbial oxidation of 1,2-dichloroethane under anoxic conditions with nitrate as electron acceptor in mixed and pure cultures. AB - Many organisms have been found to readily oxidize the prevalent contaminant 1,2 dichloroethane (1,2-DCA) to CO2 under aerobic conditions. Some organisms have also been isolated that can reduce 1,2-DCA to ethene via dihaloelimination under anaerobic, fermentative conditions. However, none have been described that can metabolize 1,2-DCA under anoxic, nitrate-reducing conditions. In microcosms prepared from aquifer material and groundwater samples from a contaminated site in eastern Louisiana, USA, 1,2-DCA was observed to degrade with nitrate as the terminal electron acceptor. Nitrate-dependent enrichment cultures were developed from these microcosms that sustained rapid 1,2-DCA degradation rates of up to 500 microM day(-1). This degradation was tightly coupled to complete reduction of nitrate via nitrite to nitrogen gas. A novel 1,2-DCA-degrading organism belonging to the Betaproteobacteria (affiliated with the genus Thauera) was isolated from this enrichment culture. However, degradation rates were much slower in cultures of the isolate than observed in the parent mixed culture. Complete mineralization of 1,2-DCA to CO2 was linked to cell growth and to nitrate reduction in both enrichment and isolated cultures. Monochloroacetate, a putative metabolite of 1,2 DCA degradation, could also be mineralized by these cultures. PMID- 16689869 TI - Trichocomaceae in bark survive high temperatures and fire. AB - Fifty-six species in the Trichocomaceae were recovered from bark of trees and shrubs from hot arid and temperate regions, and following one fire in a temperate region of Australia. Fungi were recovered from dry bark after incubation for up to 1 h at up to 105 degrees C. Fourteen species also regenerated on agar after their conidia were heated for 1 h at 105 degrees C. Anamorphic species were commonly recovered and widespread. Teleomorphic species were only recovered after heating the bark. In addition, anamorphic fungi were recovered from one plant species following a natural fire. The results support the view that both anamorphic and teleomorphic fungi may tolerate extreme temperatures in their environment while dry. PMID- 16689870 TI - In vitro decomposition of Sphagnum by some microfungi resembles white rot of wood. AB - The abilities of some ascomycetes (Myxotrichaceae) from a Sphagnum bog in Alberta to degrade cellulose, phenolics, and Sphagnum tissue were compared with those of two basidiomycetes. Most Myxotrichaceae degraded cellulose and tannic acid, and removed cell-wall components simultaneously from Sphagnum tissues, whereas the basidiomycetes degraded cellulose and insoluble phenolics, and preferentially removed the polyphenolic matrix from Sphagnum cell walls. Mass losses from Sphagnum varied from up to 50% for some ascomycetes to a maximum of 35% for the basidiomycetes. The decomposition of Sphagnum by the Myxotrichaceae was analogous to the white rot of wood and indicates that these fungi have the potential to cause significant mineralization of carbon in bogs. PMID- 16689871 TI - Selective fermentation of gentiobiose-derived oligosaccharides by human gut bacteria and influence of molecular weight. AB - Gentiooligosaccharides and alternansucrase gentiobiose acceptor products were fractionated by their degree of polymerization (DP) on a Bio-Gel P2 column. Fractions were characterized by matrix-assisted laser desorption ionization time of-flight mass spectroscopy, and incubated with human faecal bacteria under anaerobic conditions at 37 degrees C. The growth of predominant gut bacteria on the oligosaccharides was evaluated by fluorescence in situ hybridization and a prebiotic index (PI) was calculated. Lower DP gentiooligosaccharides (DP2-3) showed the highest selectivity (PI of 4.89 and 3.40, respectively), whereas DP4-5 alternansucrase gentiobiose acceptor products generated the greatest values (PI of 5.87). The production of short-chain fatty acids was also determined during the time course of the reactions. The mixture of DP6-10 alternansucrase gentiobiose acceptor products generated the highest levels of butyric acid but the lowest levels of lactic acid. Generally, for similar molecular weights, alternansucrase gentiobiose acceptor products gave higher PI values than gentiooligosaccharides. PMID- 16689872 TI - Comparison of prokaryotic diversity at offshore oceanic locations reveals a different microbiota in the Mediterranean Sea. AB - The bacterial and archaeal assemblages at two offshore sites located in polar (Greenland Sea; depth: 50 and 2000 m) and Mediterranean (Ionian Sea; depth 50 and 3000 m) waters were studied by PCR amplification and sequencing of the last 450 500 bp of the 16S rRNA gene. A total of 1621 sequences, together with alignable 16S rRNA gene fragments from the Sargasso Sea metagenome database, were analysed to ascertain variations associated with geographical location and depth. The Ionian 50 m sample appeared to be the most diverse and also had remarkable differences in terms of the prokaryotic groups retrieved; surprisingly, however, many similarities were found at the level of large-scale diversity between the Sargasso database fragments and the Greenland 50 m sample. Most sequences with more than 97% sequence similarity, a value often taken as indicative of species delimitation, were only found at a single location/depth; nevertheless, a few examples of cosmopolitan sequences were found in all samples. Depth was also an important factor and, although both deep-water samples had overall similarities, there were important differences that could be due to the warmer waters at depth of the Mediterranean Sea. PMID- 16689873 TI - Influence of dissolved organic matter source on lake bacterioplankton structure and function--implications for seasonal dynamics of community composition. AB - It has been suggested that autochthonous (internally produced) organic carbon and allochthonous (externally produced) organic carbon are utilized by phylogenetically different bacterioplankton. We examined the relationship between the source of organic matter and the structure and function of lake bacterial communities. Differences and seasonal changes in bacterial community composition in two lakes differing in their source of organic matter were followed in relation to environmental variables. We also performed batch culture experiments with amendments of various organic substrates, namely fulvic acids, leachates from algae, and birch and maple leaves. Differences in bacterial community composition between the lakes, analysed by terminal restriction fragment length polymorphism, correlated with variables related to the relative loading of autochthonous and allochthonous carbon (water colour, dissolved organic carbon, nutrients, and pH). Seasonal changes correlated with temperature, chlorophyll and dissolved organic carbon in both lakes. The substrate amendments led to differences in both structure and function, i.e. production, respiration and growth yield, of the bacterial community. In conclusion, our results suggest that the source of organic matter influences community composition both within and among lakes and that there may be a coupling between the structure and function of the bacterial community. PMID- 16689874 TI - Analysing diversity among beta-lactamase encoding genes in aquatic environments. AB - The most common mechanism of resistance to beta-lactam antibiotics is the production of beta-lactamases. These enzymes are encoded by genes that evolve rapidly, thus constituting a group characterized by high levels of molecular diversity. Most of the genetic determinants of resistance to beta-lactam antibiotics characterized until now were obtained from clinical isolates. This study was designed in order to exploit the presence of beta-lactamase gene sequences in an aquatic environment, and to get information on the distinctive features of those sequences when compared to others available on databases. DNA sequences potentially encoding proteins of three different families of clinically relevant beta-lactamases were assessed: TEM, IMP and OXA-2 derivatives. The presence of bla sequences in DNA extracted from water samples from the lagoon Ria de Aveiro was checked by PCR and hybridization. Sequences representing the three families of beta-lactamases studied were detected. The molecular diversity of the amplicons was assessed by cloning and sequence analysis, and denaturing gradient gel electrophoresis (PCR-DGGE) separation. Most of the retrieved sequences (particularly sequences representing bla(TEM)and bla(OXA-2)) were identical or very similar to beta-lactamase gene sequences previously characterized from clinical isolates. Phylogenetic analysis suggests that this aquatic ecosystem is a reservoir of molecular diverse putative bla sequences. The patterns of molecular diversity found within the beta-lactamase gene families studied do not correspond to those reported in studies focussing on clinical isolates. PMID- 16689875 TI - Soil parent material is a key determinant of the bacterial community structure in arable soils. AB - The bacterial community composition in soil and rhizosphere taken from arable field sites, differing in soil parent material and soil texture, was analyzed using terminal restriction fragment length polymorphism (T-RFLP) of 16S rRNA genes. Nine sandy to silty soils from North-East Germany could clearly be distinguished from each other, with a relatively low heterogeneity in the community structure within the field replicates. There was a relationship between the soil parent material, i.e. different glacial and aeolian sediments, and the clustering of the profiles from different sites. A site-specific grouping of T RFLP profiles was also found for the rhizosphere samples of the same field sites that were planted with potatoes. The branching of the rhizosphere profiles corresponded partly with the soil parent material, whereas the effect of the plant genotype was negligible. Selected terminal restriction fragments differing in their relative abundance within the nine soils were analyzed based on the cloning of the 16S rRNA genes of one soil sample. A high phylogenetic diversity observed to include Acidobacteria, Betaproteobacteria, Bacteroidetes, Verrucomicrobia, and Gemmatimonadetes. The assignment of three out of the seven selected terminal restriction fragments to members of Acidobacteria suggested that this group seems to participate frequently in the shifting of community structures that result from soil property changes. PMID- 16689876 TI - The influence of synthetic sheep urine on ammonia oxidizing bacterial communities in grassland soil. AB - In grazed, grassland soils, sheep urine generates heterogeneity in ammonia concentrations, with potential impact on ammonia oxidizer community structure and soil N cycling. The influence of different levels of synthetic sheep urine on ammonia oxidizers was studied in grassland soil microcosms. 'Total' and active ammonia oxidizers were distinguished by comparing denaturing gradient gel electrophoresis (DGGE) profiles following PCR and RT-PCR amplification of 16S rRNA gene fragments, targeting DNA and RNA, respectively. The RNA-based approach indicated earlier, more reproducible and finer scale qualitative shifts in ammonia oxidizing communities than DNA-based analysis, but led to amplification of a small number of nonammonia oxidizer sequences. Qualitative changes in RNA derived DGGE profiles were related to changes in nitrate accumulation. Sequence analysis of excised DGGE bands revealed that ammonia oxidizing communities in synthetic sheep urine-treated soils consisted mainly of Nitrosospira clusters 2, 3 and 4. Nitrosospira cluster 2 increased in relative abundance in microcosms treated with all levels of synthetic sheep urine. Low levels additionally led to increased relative abundance of Nitrosospira cluster 4 and medium and high levels increased relative abundance of cluster 3. Synthetic sheep urine is therefore likely to influence the spatial distribution and composition of ammonia oxidizer communities, with consequent effects on nitrate accumulation. PMID- 16689877 TI - Phylogeny of the 1-aminocyclopropane-1-carboxylic acid deaminase-encoding gene acdS in phytobeneficial and pathogenic Proteobacteria and relation with strain biogeography. AB - Deamination of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) is a key plant-beneficial trait found in plant growth-promoting rhizobacteria (PGPR) and phytosymbiotic bacteria, but the diversity of the corresponding gene (acdS) is poorly documented. Here, acdS sequences were obtained by screening putative ACC deaminase sequences listed in databases, based on phylogenetic properties and key residues. In addition, acdS was sought in 71 proteobacterial strains by PCR amplification and/or hybridization using colony dot blots. The presence of acdS was confirmed in established AcdS+ bacteria and evidenced noticeably in Azospirillum (previously reported as AcdS-), in 10 species of Burkholderia and six Burkholderia cepacia genomovars (which included PGPR, phytopathogens and opportunistic human pathogens), and in five Agrobacterium genomovars. The occurrence of acdS in true and opportunistic pathogens raises new questions concerning their ecology in plant-associated habitats. Many (but not all) acdS+ bacteria displayed ACC deaminase activity in vitro, including two Burkholderia clinical isolates. Phylogenetic analysis of partial acdS and deduced AcdS sequences evidenced three main phylogenetic clusters, each gathering pathogens and plant-beneficial strains of contrasting geographic and habitat origins. The acdS phylogenetic tree was only partly congruent with the rrs tree. Two clusters gathered both Betaprotobacteria and Gammaproteobacteria, suggesting extensive horizontal transfers of acdS, noticeably between plant-associated Proteobacteria. PMID- 16689878 TI - Characterization of bacterial populations recovered from the teat canals of lactating dairy and beef cattle by 16S rRNA gene sequence analysis. AB - Bovine mastitis is of major concern to the dairy industry worldwide. The bovine teat canal is the primary route through which pathogens enter the mammary gland. The microflora of the teat canals of dairy and beef cattle was investigated by analysis of 16S rRNA gene sequences. The goal was to examine the global difference between dairy cattle, which are sanitized on a regular basis, and beef cattle, which receive little sanitary management. A diverse population of 16S rRNA gene sequences was recovered from both the dairy and the beef herd samples, with diversity higher in the beef sample. Analysis revealed the presence of 90 operational taxonomic units (OTUs) among 156 sequences, with 45 OTUs in the dairy sample and 53 OTUs in the beef sample. Only eight OTUs were common to both samples. Members of the classes Clostridia and Bacilli dominated both samples, followed by Actinobacteria and Proteobacteria. The dairy sample contained a cluster (20/80 clones) of Staphylococcaceae members, seven of which were identifiable as coagulase-negative Staphylococcus species. The beef sample was dominated by members of the genus Clostridia, many of which have not been previously cultured. The results suggest that the microorganisms present in the bovine teat canal are more diverse than previously described. PMID- 16689879 TI - Comparison of PCR primer-based strategies for characterization of ammonia oxidizer communities in environmental samples. AB - PCR-based techniques are commonly used to characterize microbial communities, but are subject to bias that is difficult to assess. This study aimed to evaluate bias of several PCR primer-based strategies used to study diversity of autotrophic ammonia oxidizers. 16S rRNA genes from soil- or sediment-DNA were amplified using primers considered either selective or specific for betaproteobacterial ammonia oxidizers. Five approaches were assessed: (a) amplification with primers betaAMO143f-betaAMO1315r; (b) amplification with primers CTO189f-CTO654r; (c) nested amplification with betaAMO143f-betaAMO1315r followed by CTO189f-CTO654r primers; (d) nested amplification with betaAMO143f betaAMO1315r and CTO189f-Pf1053r primers; (e) nested amplification with 27f-1492r and CTO189f-CTO654r primers. Amplification products were characterized by denaturing gradient gel electrophoresis (DGGE) analysis after further amplification with 357f-GC-518r primers. DGGE profiles of soil communities were heterogeneous and depended on the approach followed. Ammonia oxidizer diversity was higher using approaches (b), (c) and (e) than using (a) and (d), where sequences of the most prominent bands showed similarities to nonammonia oxidizers. Profiles from marine sediments were more consistent, regardless of the approach adopted, and sequence analysis of excised bands indicated that these consisted of ammonia oxidizers only. The study demonstrates the importance of choice of primer, of screening for sequences of nontarget organisms and use of several approaches when characterizing microbial communities in natural environments. PMID- 16689880 TI - The endometrium of the anoestrous female pig: studies on infiltration by cells of the immune system. AB - The aim of this study was to investigate the distribution of immune cells in the endometrium of anoestrous female pigs, five sows in anoestrus by lactation and five pre-pubertal gilts (Swedish Landrace x Swedish Yorkshire). Uterine samples, taken immediately after slaughter, were fixed, embedded in plastic resin and stained with toluidine blue or cryo fixed and stored in a freezer at -70 degrees C until analysed by immunohistochemistry with an avidin-biotin peroxidase method. Immune cells in the surface (luminal) and the glandular epithelium as well as the subepithelial and the glandular connective tissue layers were counted using light microscopy. In the surface (luminal) and the glandular epithelia of gilts and sows, lymphocytes were the predominant immune cells found. There were no significant differences between gilts and sows. Macrophages were detected in the glandular epithelium of sows but not in gilts. In the subepithelial and the glandular connective tissue layers of both gilts and sows, lymphocytes were also the most common immune cells found. The numbers of lymphocytes and macrophages were significantly higher in the sows than in the gilts (p 1000 embryos) from a approximately 80 m linear transect in Indiana, USA. Embryos were genotyped at four variable microsatellite loci and the resulting progeny array data were used to reconstruct multilocus genotypes of the parental dams and sires for each egg mass. UPGMA analysis of genetic distances among embryos resolved four instances of egg mass admixture, where two or more females had oviposited at exactly the same site resulting in the mixing of independent cohorts. In total, 41 discrete egg masses were available for parentage analyses. Twenty-three egg masses (56%) consisted exclusively of full siblings (i.e. were singly sired) and 18 (44%) were multiply sired (mean 2.6 males/clutch). Parentage could be genetically assigned to one of 17 distinct parent pairs involving at least 15 females and 14 different males. Reproductive skew was evident among males who sired multiply sired clutches. Additional evidence of the effects of sexual selection on male reproductive success was apparent via significant positive correlations between male mating and reproductive success. Females frequently partitioned their clutches into multiple discrete egg masses that were separated from one another by as many as 43 m. Collectively, these data provide the first direct evidence for polygynandry in a wild population of tiger salamanders. PMID- 16689908 TI - Parentage analysis detects cryptic precapture dispersal in a philopatric rodent. AB - Locating birthplaces using genetic parentage determination can increase the precision and accuracy with which animal dispersal patterns are established. We re-analyse patterns of movement away from the birthplace as a function of time, sex and population density for a sample of 303 banner-tailed kangaroo rats, Dipodomys spectabilis. We located birth sites using a combination of likelihood based parentage analysis with live-trapping of mothers during the breeding season. The results demonstrate that natal-breeding site distances are density dependent in this species; in particular, both sexes emigrate earlier in the year, and females disperse farther than males, at low population densities. Banner-tailed kangaroo rats were chosen as a study system because live-trapping easily detects maternal and offspring locations; nevertheless, parentage analysis reveals that some offspring evade early detection and move substantial distances before their first capture. In a few cases, the approach even detects dispersal out of the natal 'deme' prior to first capture. Parentage analysis confirms the extreme philopatry of both sexes but indicates that prior estimates of median dispersal distance were too low. For D. spectabilis, more accurate location of individual birthplaces clarifies patterns of sex bias and density dependence in dispersal, and may resolve apparent discrepancies between direct and indirect estimates of dispersal distance. For species in which mothers can be more reliably trapped than juveniles, using offspring genotypes to locate parents is a novel way that genetic techniques can contribute to the analysis of animal dispersal. PMID- 16689909 TI - Finding fathers: spatio-temporal analysis of paternity assignment in grey seals (Halichoerus grypus). AB - Molecular studies of pinniped breeding systems exhibit a broad range of agreement and disagreement with observational indices of male breeding success. Grey seal studies have reported considerable discrepancies between genetic and behavioural paternity measures that have been interpreted as evidence of previously unidentified male strategies and/or tactics. Therefore, these studies have the power to fundamentally alter our perceptions of mating systems. However, other pinniped studies exhibit no such disagreements, and one possible explanation for disparities may be sampling biases in space and time. Therefore, it is essential that potential sampling biases are examined to evaluate the likelihood of previously unidentified male strategies. We examined paternities assigned at the North Rona grey seal colony between 1999 and 2002 in relation to concurrent detailed behavioural and locational data for males and females. We found that (i) for females observed in sexual interaction(s) during their oestrus period, it was highly probable that one of the interacting males fathered their next pup; (ii) over 80% of assigned paternities agreed with observations of the in-colony behaviour and spatio-temporal proximity of the males and females involved; and (iii) a minority of females exhibit mate choice and seek sires outside their local male's home range, although evidence suggests that these females mate on the colony rather than at sea. In conclusion, nearly all paternities assigned agreed with expectation based upon detailed knowledge of the spatio-temporal patterns of individuals during the breeding season. We found little evidence of unidentified male strategies at North Rona, Scotland, whereas further examination of mechanisms of female choice may be productive. PMID- 16689910 TI - A fungal endophyte of black spruce (Picea mariana) needles is also an aquatic hyphomycete. AB - An aquatic hyphomycete, Dwayaangam sp., was isolated from superficially sterilized black spruce (Picea mariana) needles submerged in aerated water in a small glass chamber (microcosm). The internal transcribed spacer (ITS) sequence of this fungus and of a commonly encountered foliar endophyte isolated from P. mariana showed a high degree of similarity. When sporulation was induced in the microcosm, both the aquatic hyphomycete and the endophyte isolate produced similar aquatic conidia after 30 days, which is longer than previously documented in similar studies. Without the use of molecular tools, the link between the aquatic and endophytic phases of the fungus would have gone unnoticed. This is the first time that a fungal endophyte of conifer needles has been shown to have an aquatic phase. Its presence both as a foliar endophyte and a sporulating aquatic fungus suggests an alternating life cycle between the two environments. PMID- 16689911 TI - Molecular detection of predation by soil micro-arthropods on nematodes. AB - The relative importance of the factors driving change in the population dynamics of nematodes in the soil is almost completely unknown. Top-down control by micro arthropod predators may have a significant impact on nematode population dynamics. We report experiments showing that mites and Collembola were capable of reducing nematode numbers in the laboratory and were feeding on a targeted nematode species in the field. A PCR-based approach was developed for the detection of predation on three species of slug- and insect-pathogenic nematodes: Phasmarhabditis hermaphrodita, Heterorhabditis megidis and Steinernema feltiae. The collembolan Folsomia candida and the mesostigmatid mite Stratiolaelaps miles were employed as model predators to calibrate post-ingestion prey DNA detection times. Fragments of cytochrome oxidase I (COI) mtDNA were sequenced and species specific primers were designed, amplifying 154-, 154- and 203-bp fragments for each of the nematode species. Detection times for nematode DNA within the guts of Collembola were longer than in mites, with half-lives (50% of samples testing positive) of 08.75 h and 05.03 h, respectively. F. candida significantly reduced numbers of the nematode H. megidis, with rates of predation of approximately 0.4 nematode infective juveniles per collembolan per hour over 10 h. Four taxa of field-caught micro-arthropod that had been exposed to the nematode P. hermaphrodita for a period of 12 h were analysed and significant numbers of three taxa tested positive. This is the first application of PCR techniques for the study of nematophagy and the first time these techniques have been used to measure predation on nematodes in the field. PMID- 16689912 TI - Contrasting effects of heterozygosity on survival and hookworm resistance in California sea lion pups. AB - Low genetic heterozygosity is associated with loss of fitness in many natural populations. However, it remains unclear whether the mechanism is related to general (i.e. inbreeding) or local effects, in particular from a subset of loci lying close to genes under balancing selection. Here we analyse involving heterozygosity-fitness correlations on neonatal survival of California sea lions and on susceptibility to hookworm (Uncinaria spp.) infection, the single most important cause of pup mortality. We show that regardless of differences in hookworm burden, homozygosity is a key predictor of hookworm-related lesions, with no single locus contributing disproportionately. Conversely, the subsequent occurrence of anaemia due to blood loss in infected pups is overwhelmingly associated with homozygosity at one particular locus, all other loci showing no pattern. Our results suggest contrasting genetic mechanisms underlying two pathologies related to the same pathogen. First, relatively inbred pups are less able to expel hookworms and prevent their attachment to the intestinal mucosa, possibly due to a weakened immune response. In contrast, infected pups that are homozygous for a gene near to microsatellite Hg4.2 are strongly predisposed to anaemia. As yet, this gene is unknown, but could plausibly be involved in the blood-coagulation cascade. Taken together, these results suggest that pathogenic burden alone may not be the main factor regulating pathogen-related mortality in natural populations. Our study could have important implications for the conservation of small, isolated or threatened populations, particularly when they are at a risk of facing pathogenic challenges. PMID- 16689913 TI - When invaders meet locally adapted types: rapid moulding of hybrid zones between sculpins (Cottus, Pisces) in the Rhine system. AB - Hybrid zones are commonly studied to dissect the processes that drive divergence among lineages, which have incomplete barriers of reproduction. Most hybrid zones have existed for an extended time making inferences on the initial mode of formation difficult. It is a priori unclear how fast a nascent hybrid zone would form as a response to endogenous and exogenous factors. We have studied several hybrid zones between two lineages of sculpins (Cottus spp.), which emerged due to a recent range expansion of one of the lineages along the river Rhine in the early 1990s. Applying a dense sampling across two contact areas and using a highly informative set of 45 microsatellite markers we found pronounced genetic structure. Steep genetic clines suggest that strong selective forces have shaped the respective hybrid zones from the beginning. We find that the zones are coupled to ecological transitions from small streams to larger rivers. The width of these zones is much smaller than estimates of annual individual dispersal distances, as estimated outside of the hybrid zones. The pattern is apparently not strongly affected by pre- or postzygotic reproductive isolation because numerous backcross hybrids occur within the zones. This suggests that strong natural selection acts against immigrant genotypes. The study exemplifies how local adaptation can play a key role in preventing admixture in dependence of the ecological context. PMID- 16689914 TI - No relationship between microsatellite variation and neonatal fitness in Antarctic fur seals, Arctocephalus gazella. AB - Published studies of wild vertebrate populations have almost universally reported positive associations between genetic variation measured at microsatellite loci and fitness, creating the impression of ubiquity both in terms of the species and the traits involved. However, there is concern that this picture may be misleading because negative results frequently go unpublished. Here, we analyse the relationship between genotypic variation at nine highly variable microsatellite loci and neonatal fitness in 1070 Antarctic fur seal pups born at Bird Island, South Georgia. Despite our relatively large sample size, we find no significant association between three different measures of heterozygosity and two fitness traits, birth weight and survival. Furthermore, increasing genetic resolution by calculating parental relatedness also yields no association between genetic variation and fitness. Our findings are consistent with necropsy data showing that most pups die from starvation or trauma, conditions that are unlikely to be influenced strongly by genetic factors, particularly if the benefits of high heterozygosity are linked to immune-related genes. PMID- 16689915 TI - Shared alleles in sympatric oaks: recurrent gene flow is a more parsimonious explanation than ancestral polymorphism. PMID- 16689916 TI - Mainstreaming risk reduction in urban planning and housing: a challenge for international aid organisations. AB - The effects of 'natural' disasters in cities can be worse than in other environments, with poor and marginalised urban communities in the developing world being most at risk. To avoid post-disaster destruction and the forced eviction of these communities, proactive and preventive urban planning, including housing, is required. This paper examines current perceptions and practices within international aid organisations regarding the existing and potential roles of urban planning as a tool for reducing disaster risk. It reveals that urban planning confronts many of the generic challenges to mainstreaming risk reduction in development planning. However, it faces additional barriers. The main reasons for the identified lack of integration of urban planning and risk reduction are, first, the marginal position of both fields within international aid organisations, and second, an incompatibility between the respective professional disciplines. To achieve better integration, a conceptual shift from conventional to non-traditional urban planning is proposed. This paper suggests related operative measures and initiatives to achieve this change. PMID- 16689917 TI - Psychological assessment of children in disasters and emergencies. AB - Children and adolescents are among the most vulnerable members of communities affected by disasters and emergencies. There is a tremendous need for a systematic post-disaster psychological assessment of children and adolescents in order to understand better post-traumatic symptomatology in children and to identify populations that require an early intervention. This article reviews psychological instruments that are suitable for screening children and adolescents in emergency and disaster contexts for four different types of post traumatic responses: post-traumatic stress disorder; depression; anxiety disorders; and behavioural disorders. A description of each instrument and psychometric data are provided, along with recommendations on the most appropriate instruments to be utilised in different emergency environments and a summary of previous post-disaster evaluations that have used each type. In addition to selecting apposite instruments, other important issues that should be taken into account when conducting post-emergency mental health needs appraisals of children and adolescents are discussed. PMID- 16689918 TI - Using design effects from previous cluster surveys to guide sample size calculation in emergency settings. AB - A good estimate of the design effect is critical for calculating the most efficient sample size for cluster surveys. We reviewed the design effects for seven nutrition and health outcomes from nine population-based cluster surveys conducted in emergency settings. Most of the design effects for outcomes in children, and one-half of the design effects for crude mortality, were below two. A reassessment of mortality data from Kosovo and Badghis, Afghanistan revealed that, given the same number of clusters, changing sample size had a relatively small impact on the precision of the estimate of mortality. We concluded that, in most surveys, assuming a design effect of 1.5 for acute malnutrition in children and two or less for crude mortality would produce a more efficient sample size. In addition, enhancing the sample size in cluster surveys without increasing the number of clusters may not result in substantial improvements in precision. PMID- 16689919 TI - Organisational learning and self-adaptation in dynamic disaster environments. AB - This paper examines the problems associated with inter-organisational learning and adaptation in the dynamic environments that characterise disasters. The research uses both qualitative and quantitative methods to investigate whether organisational learning took place during and in the time in between five disaster response operations in Turkey. The availability of information and its exchange and distribution within and among organisational actors determine whether self-adaptation happens in the course of a disaster response operation. Organisational flexibility supported by an appropriate information infrastructure creates conditions conducive to essential interaction and permits the flow of information. The study found that no significant organisational learning occurred within Turkish disaster management following the earthquakes in Erzincan (1992), Dinar (1995) and Ceyhan (1998). By contrast, the 'symmetry-breaking' Marmara earthquake of 1999 initiated a 'double loop' learning process that led to change in the organisational, technical and cultural aspects of Turkish disaster management, as revealed by the Duzce earthquake response operations. PMID- 16689920 TI - About bridges and bonds: community responses to the 2000 floods in Mabalane district, Mozambique. AB - Massive floods hit Mozambique in 2000.1 Many of the affected regions benefited from external aid. This paper investigates how the floods impacted on two communities in the interior of the Limpopo Valley, which did not receive significant assistance during the event and in the immediate aftermath. Drawing on the livelihood approach, it focuses on the role of two types of local resources: knowledge and social capital. The paper shows that the scale of the 2000 floods surpassed the response capacity of these resources and looks at how wealth and gender influenced access by households to the local support mechanisms that enhance survival and recovery. It appears that the floods mainly affected wealthier households, as these more frequently lost houses and cattle. The paper points to the importance of ploughs and cattle for local support mechanisms, which, in turn, most likely reduced the capacities of communities to recover from the disaster. PMID- 16689921 TI - Economic evaluation of measles catch-up and follow-up campaigns in Afghanistan in 2002 and 2003. AB - This paper assesses the cost-effectiveness of, and the return on the investment in, the 2002 catch-up and the 2003 follow-up measles campaigns in Afghanistan from the perspective of the donor. The catch-up campaign targeted nearly 12 million children aged between six months and 12 years, while the follow-up campaign targeted over five million children aged between 9 and 59 months. Both campaigns successfully vaccinated approximately 96 per cent of the respective target populations, and are expected to avert an estimated 301,000 measles deaths over the next 10 years. The average cost per dose of measles vaccine delivered was USD 0.40. The cost per death prevented is USD 23.6, assuming a case fatality rate of 10 per cent and a discount rate of three per cent. With more than 42,000 measles deaths avoided for every one million US dollars spent, the campaigns are an excellent public health investment for precluding childhood mortality in a country affected by a complex emergency. PMID- 16689922 TI - Adverse events following immunization attributable to measles vaccine are rare. PMID- 16689923 TI - Protein-misfolding diseases and chaperone-based therapeutic approaches. AB - A large number of neurodegenerative diseases in humans result from protein misfolding and aggregation. Protein misfolding is believed to be the primary cause of Alzheimer's disease, Parkinson's disease, Huntington's disease, Creutzfeldt-Jakob disease, cystic fibrosis, Gaucher's disease and many other degenerative and neurodegenerative disorders. Cellular molecular chaperones, which are ubiquitous, stress-induced proteins, and newly found chemical and pharmacological chaperones have been found to be effective in preventing misfolding of different disease-causing proteins, essentially reducing the severity of several neurodegenerative disorders and many other protein-misfolding diseases. In this review, we discuss the probable mechanisms of several protein misfolding diseases in humans, as well as therapeutic approaches for countering them. The role of molecular, chemical and pharmacological chaperones in suppressing the effect of protein misfolding-induced consequences in humans is explained in detail. Functional aspects of the different types of chaperones suggest their uses as potential therapeutic agents against different types of degenerative diseases, including neurodegenerative disorders. PMID- 16689924 TI - Interaction of G-rich GT oligonucleotides with nuclear-associated eEF1A is correlated with their antiproliferative effect in haematopoietic human cancer cell lines. AB - G-rich GT oligonucleotides with a different content of G clusters have been evaluated for their ability to exert cytotoxicity and to bind to nuclear associated proteins in T-lymphoblast CCRF-CEM cells. Only the oligomers that did not form G-based structures or had a poor structure, under physiological conditions, were able to exert significant cellular growth inhibition effect. The cytotoxicity of these oligomers was related to their binding to the nuclear associated eEF1A protein, but not to the recognition of nucleolin or other proteins. In particular, GT oligomers adopting a conformation compatible with G quadruplex, did not exert cytotoxicity and did not bind to eEF1A. The overall results suggest that the ability of oligomers to adopt a G-quadruplex-type secondary structure in a physiological buffer containing 150 mM NaCl is not a prerequisite for antiproliferative effect in haematopoietic cancer cells. The cytotoxicity of G-rich GT oligomers was shown to be tightly related to their binding affinity for eEF1A protein. PMID- 16689925 TI - Benzo[a]pyrene impairs beta-adrenergic stimulation of adipose tissue lipolysis and causes weight gain in mice. A novel molecular mechanism of toxicity for a common food pollutant. AB - Benzo[a]pyrene (B[a]P) is a common food pollutant that causes DNA adduct formation and is carcinogenic. The report of a positive correlation between human plasma B[a]P levels and body mass index, together with B[a]P's lipophilicity, led us to test for possible adverse effects of B[a]P on adipose tissue. In ex vivo experiments using primary murine adipocytes, B[a]P rapidly (within minutes) and directly inhibited epinephrine-induced lipolysis (up to 75%) in a dose-dependent manner. Half-maximum inhibition was obtained with a B[a]P concentration of 0.9 mg.L(-1) (3.5 microm). Lipolysis induced by beta(1)-, beta(2)- and beta(3) adrenoreceptor-specific agonists, as well as ACTH, were also significantly inhibited by B[a]P, whereas forskolin-induced lipolysis was not B[a]P-sensitive. Similar inhibition of catecholamine-induced lipolysis by B[a]P was also seen in isolated human adipocytes; half-maximum inhibition of lipolysis was achieved with a B[a]P concentration of 0.02 mg.L(-1) (0.08 microm). In vivo treatment of C57Bl/6J mice with 0.4 mg.kg(-1) B[a]P inhibited epinephrine-induced release of free fatty acids by 70%. Chronic exposure of mice to B[a]P (0.5 mg.kg(-1) injected i.p. every 48 h) for 15 days also decreased lipolytic response to epinephrine and induced a 43% higher weight gain compared with controls (B[a]P: 2.23 +/- 0.12 g versus control: 1.56 +/- 0.18 g, P < 0.01) due to increased fat mass. The weight gain occurred consistently without detectable changes in food intake. These results reveal a novel molecular mechanism of toxicity for the environmental pollutant B[a]P and introduce the notion that chronic exposure of human population to B[a]P and possibly other polycyclic aromatic hydrocarbons could have an impact on metabolic disorders, such as obesity. PMID- 16689926 TI - alpha-Conotoxin GI benzoylphenylalanine derivatives. (1)H-NMR structures and photoaffinity labeling of the Torpedo californica nicotinic acetylcholine receptor. AB - alpha-Conotoxins are small peptides from cone snail venoms that function as nicotinic acetylcholine receptor (nAChR)-competitive antagonists differentiating between nAChR subtypes. Current understanding about the mechanism of these selective interactions is based largely on mutational analyses, which identify amino acids in the toxin and nAChR that determine the energetics of ligand binding. To identify regions of the nAChR involved in alpha-conotoxin binding by use of photoactivated cross-linking, two benzoylphenylalanine (Bpa) analogs of alpha-conotoxin GI, GI(Bpa12) and GI(Bpa4), were synthesized by replacing the respective residues with Bpa, and their (1)H-NMR structures were determined. Both analogs preserved the GI conformation, but only GI(Bpa12) displaced (125)I labeled GI from the Torpedo californica nAChR. (125)I-labeled GI(Bpa12) bound to two sites on the receptor (K(d) 13 and 1800 nM), and on UV irradiation specifically photolabeled the alpha, gamma and delta subunits. Photolabeling sites were mapped by selective proteolysis and enzymatic deglycosylation, combined with SDS/PAGE, HPLC and Edman degradation. In the alpha subunit, cobratoxin-inhibited incorporation was limited to the 22-kDa fragment beginning at alphaSer173 and containing the agonist-binding site segment C. In the gamma subunit, radioactivity was localized to two distinct peptides containing agonist binding site segments F and D: nonglycosylated 24-kDa and glycosylated 13-kDa fragments starting at gammaAla167 and gammaAla49, respectively. The labeling of these fragments is discussed in terms of a model of GI(Bpa12) bound to the extracellular domain of the Torpedo nAChR homology model derived from the cryo electron microscopy structure of Torpedo marmorata nAChR and X-ray crystal structures of snail acetylcholine-binding protein complexes with agonists and antagonists. PMID- 16689927 TI - Effects of sphingomyelin, cholesterol and zinc ions on the binding, insertion and aggregation of the amyloid Abeta(1-40) peptide in solid-supported lipid bilayers. AB - We utilized plasmon-waveguide resonance (PWR) spectroscopy to follow the effects of sphingomyelin, cholesterol and zinc ions on the binding and aggregation of the amyloid beta peptide(1-40) in lipid bilayers. With a dioleoylphosphatidylcholine (DOPC) bilayer, peptide binding was observed, but no aggregation occurred over a period of 15 h. In contrast, similar binding was found with a brain sphingomyelin (SM) bilayer, but in this case an exponential aggregation process was observed during the same time interval. When the SM bilayer included 35% cholesterol, an increase of approximately 2.5-fold occurred in the amount of peptide bound, with a similar increase in the extent of aggregation, the latter resulting in decreases in the bilayer packing density and displacement of lipid. Peptide association with a bilayer formed from equimolar amounts of DOPC, SM and cholesterol was followed using a high-resolution PWR sensor that allowed microdomains to be observed. Biphasic binding to both domains occurred, but predominantly to the SM-rich domain, initially to the surface and at higher peptide concentrations within the interior of the bilayer. Again, aggregation was observed and occurred within both microdomains, resulting in lipid displacement. We attribute the aggregation in the DOPC-enriched domain to be a consequence of lipid mixing within these microdomains, resulting in the presence of small amounts of SM and cholesterol in the DOPC microdomain. When 1 mM zinc was present, an increase of approximately threefold in the amount of peptide association was observed, as well as large changes in mass and bilayer structure as a consequence of peptide aggregation, occurring without loss of bilayer integrity. A structural interpretation of peptide interaction with the bilayer is presented based on the results of simulation analysis of the PWR spectra. PMID- 16689928 TI - Alcohol linked to enhanced angiogenesis in rat model of choroidal neovascularization. AB - One of the pathologic complications of exudative (i.e. wet-type) age-related macular degeneration (AMD) is choroidal neovascularization (CNV). The aim of this study was to investigate whether chronic and heavy alcohol consumption influenced the development of CNV in a rat model. The oxidative metabolism of alcohol is minimal or absent in the eye, so that ethanol is metabolized via a nonoxidative pathway to form fatty acid ethyl esters (FAEE). Fatty acid ethyl ester synthase (FAEES) was purified from the choroid of Brown Norway (BN) rats. The purified protein was 60 kDa in size and the antibody raised against this protein showed a single band on western blot. BN rats on a regular diet were fed alcohol for 10 weeks. Control rats were fed water with a regular diet and pair-fed control rats were fed regular diet, water and glucose. We found that FAEES activity was increased 4.0-fold in the choroid of alcohol-treated rats compared with controls. The amount of ethyl esters produced in the choroid of 10 week alcohol-fed rats was 7.4-fold more than rats fed alcohol for 1 week. The increased accumulation of ethyl esters was associated with a 3.0-fold increased expression of cyclin E and cyclin E/CDK2; however, the level of the cyclin kinase inhibitor, p27Kip, did not change. The increased accumulation of ethyl esters was also associated with 3.0 fold decreased expression of APN in the choroid. We also found that the size of CNV increased by 28% in alcohol-fed rats. Thus, our study showed that chronic, heavy alcohol intake was associated with both an increased accumulation of ethyl esters in the choroid and an exacerbation of the CNV induced by laser treatment. These results may provide insight into the link between heavy alcohol consumption and exudative AMD. PMID- 16689929 TI - Probing protein-chromophore interactions in Cph1 phytochrome by mutagenesis. AB - We have investigated mutants of phytochrome Cph1 from the cyanobacterium Synechocystis PCC6803 in order to study chromophore-protein interactions. Cph1Delta2, the 514-residue N-terminal sensor module produced as a recombinant His6-tagged apoprotein in Escherichia coli, autoassembles in vitro to form a holoprotein photochemically indistinguishable from the full-length product. We generated 12 site-directed mutants of Cph1Delta2, focusing on conserved residues which might be involved in chromophore-protein autoassembly and photoconversion. Folding, phycocyanobilin-binding and Pr-->Pfr photoconversion were analysed using CD and UV-visible spectroscopy. MALDI-TOF-MS confirmed C259 as the chromophore attachment site. C259L is unable to attach the chromophore covalently but still autoassembles to form a red-shifted photochromic holoprotein. H260Q shows UV visible properties similar to the wild-type at pH 7.0 but both Pr and Pfr (reversibly) bleach at pH 9.0, indicating that the imidazole side chain buffers chromophore protonation. Mutations at E189 disturbed folding but the residue is not essential for chromophore-protein autoassembly. In D207A, whereas red irradiation of the ground state leads to bleaching of the red Pr band as in the wild-type, a Pfr-like peak does not arise, implicating D207 as a proton donor for a deprotonated intermediate prior to Pfr. UV-Vis spectra of both H260Q under alkaline conditions and D207A point to a particular significance of protonation in the Pfr state, possibly implying proton migration (release and re-uptake) during Pr-->Pfr photoconversion. The findings are discussed in relation to the recently published 3D structure of a bacteriophytochrome fragment. PMID- 16689930 TI - Gradual phosphorylation regulates PC4 coactivator function. AB - The unstructured N-terminal domain of the transcriptional cofactor PC4 contains multiple phosphorylation sites that regulate activity. The phosphorylation status differentially influences the various biochemical functions performed by the structured core of PC4. Binding to ssDNA is slightly enhanced by phosphorylation of one serine residue, which is not augmented by further phosphorylation. The presence of at least two phosphoserines decreases DNA-unwinding activity and abrogates binding to the transcriptional activator VP16. Phosphorylation gradually decreases the binding affinity for dsDNA. These phosphorylation dependent changes in PC4 activities correlate with the sequential functions PC4 fulfils throughout the transcription cycle. MS and NMR revealed that up to eight serines are progressively phosphorylated towards the N-terminus, resulting in gradual environmental changes in the C-terminal direction of the following lysine rich region. Also within the structured core, primarily around the interaction surfaces, environmental changes are observed. We propose a model for co-ordinated changes in PC4 cofactor functions, mediated by phosphorylation status-dependent gradual masking of the lysine-rich region causing shielding or exposure of interaction surfaces. PMID- 16689931 TI - Mycobacterium tuberculosis H37Rv ESAT-6-CFP-10 complex formation confers thermodynamic and biochemical stability. AB - The 6-kDa early secretory antigenic target (ESAT-6) and culture filtrate protein 10 (CFP-10), expressed from the region of deletion-1 (RD1) of Mycobacterium tuberculosis H37Rv, are known to play a key role in virulence. In this study, we explored the thermodynamic and biochemical changes associated with the formation of the 1 : 1 heterodimeric complex between ESAT-6 and CFP-10. Using isothermal titration calorimetry (ITC), we precisely determined the association constant and free energy change for formation of the complex to be 2 x 10(7) M(-1) and -9.95 kcal.mol(-1), respectively. Strikingly, the thermal unfolding of the ESAT-6-CFP 10 heterodimeric complex was completely reversible, with a T(m) of 53.4 degrees C and DeltaH of 69 kcal.mol(-1). Mixing of ESAT-6 and CFP-10 at any temperature below the T(m) of the complex led to induction of helical conformation, suggesting molecular recognition between specific segments of unfolded ESAT-6 and CFP-10. Enhanced biochemical stability of the complex was indicated by protection of ESAT-6 and an N-terminal fragment of CFP-10 from proteolysis with trypsin. However, the flexible C-terminal of CFP-10 in the complex, which has been shown to be responsible for binding to macrophages and monocytes, was cleaved by trypsin. In the presence of phospholipid membranes, ESAT-6, but not CFP-10 and the complex, showed an increase in alpha-helical content and enhanced thermal stability. Overall, complex formation resulted in structural changes, enhanced thermodynamic and biochemical stability, and loss of binding to phospholipid membranes. These features of complex formation probably determine the physiological role of ESAT-6, CFP-10 and/or the complex in vivo. The ITC and thermal unfolding approach described in this study can readily be applied to characterization of the 11 other pairs of ESAT-6 family proteins and for screening ESAT-6 and CFP-10 mutants. PMID- 16689932 TI - Dissecting the role of protein-protein and protein-nucleic acid interactions in MS2 bacteriophage stability. AB - To investigate the role of protein-protein and protein-nucleic acid interactions in virus assembly, we compared the stabilities of native bacteriophage MS2, virus like particles (VLPs) containing nonviral RNAs, and an assembly-defective coat protein mutant (dlFG) and its single-chain variant (sc-dlFG). Physical (high pressure) and chemical (urea and guanidine hydrochloride) agents were used to promote virus disassembly and protein denaturation, and the changes in virus and protein structure were monitored by measuring tryptophan intrinsic fluorescence, bis-ANS probe fluorescence, and light scattering. We found that VLPs dissociate into capsid proteins that remain folded and more stable than the proteins dissociated from authentic particles. The proposed model is that the capsid disassembles but the protein remains bound to the heterologous RNA encased by VLPs. The dlFG dimerizes correctly, but fails to assemble into capsids, because it lacks the 15-amino acid FG loop involved in inter-dimer interactions at the viral fivefold and quasi-sixfold axes. This protein was very unstable and, when compared with the dissociation/denaturation of the VLPs and the wild-type virus, it was much more susceptible to chemical and physical perturbation. Genetic fusion of the two subunits of the dimer in the single-chain dimer sc-dlFG stabilized the protein, as did the presence of 34-bp poly(GC) DNA. These studies reveal mechanisms by which interactions in the capsid lattice can be sufficiently stable and specific to ensure assembly, and they shed light on the processes that lead to the formation of infectious viral particles. PMID- 16689933 TI - Regional and segmental flexibility of antibodies in interaction with antigens of different size. AB - The interaction of antibodies (Abs) with protein antigens (Ags) of different size, such as hen egg white lysozyme, ovalbumin, and bovine serum albumin, was examined using analytical ultracentrifugation, electrospray ionization time-of flight mass spectrometry, and surface plasmon resonance in order to estimate regional and segmental Ab flexibility. When both Abs and Ags were free in solution, sedimentation equilibrium and surface plasmon resonance analyses showed the formation of an Ag(2)Ab(1) complexes regardless of Ag size, suggesting that the Fab arms were able to move to avoid interference between Ags bound to Ab combining sites. The Ag(2)Ab(1) complex, as well as the Ag(1)Ab(1) complex, was observed by MS. However, when Abs were immobilized on the surface of a sensor chip through the Fc region, the stoichiometry of the Ag-Ab complex was dependent on the Ag size; Ag(2)Ab(1) forming with hen egg white lysozyme and Ag(1)Ab(1) with ovalbumin and bovine serum albumin. These results indicated that immobilization of the Fc region reduces the dynamic range of the Fab arms and results in interference from the first Ag bound to either combining site, which in turn prevents the binding of the second Ag to the other combining site. Our results allow us to propose that the Fab arms of B-cell receptors whose Fc regions are immobilized on cell surface have a reduced dynamic range. PMID- 16689934 TI - Inorganic phosphate regulates the binding of cofilin to actin filaments. AB - Inorganic phosphate (Pi) and cofilin/actin depolymerizing factor proteins have opposite effects on actin filament structure and dynamics. Pi stabilizes the subdomain 2 in F-actin and decreases the critical concentration for actin polymerization. Conversely, cofilin enhances disorder in subdomain 2, increases the critical concentration, and accelerates actin treadmilling. Here, we report that Pi inhibits the rate, but not the extent of cofilin binding to actin filaments. This inhibition is also significant at physiological concentrations of Pi, and more pronounced at low pH. Cofilin prevents conformational changes in F actin induced by Pi, even at high Pi concentrations, probably because allosteric changes in the nucleotide cleft decrease the affinity of Pi to F-actin. Cofilin induced allosteric changes in the nucleotide cleft of F-actin are also indicated by an increase in fluorescence emission and a decrease in the accessibility of etheno-ADP to collisional quenchers. These changes transform the nucleotide cleft of F-actin to G-actin-like. Pi regulation of cofilin binding and the cofilin regulation of Pi binding to F-actin can be important aspects of actin based cell motility. PMID- 16689935 TI - DNA strand exchange activity of rice recombinase OsDmc1 monitored by fluorescence resonance energy transfer and the role of ATP hydrolysis. AB - Rad51 and disrupted meiotic cDNA1 (Dmc1) are the two eukaryotic DNA recombinases that participate in homology search and strand exchange reactions during homologous recombination mediated DNA repair. Rad51 expresses in both mitotic and meiotic tissues whereas Dmc1 is confined to meiosis. DNA binding and pairing activities of Oryza sativa disrupted meiotic cDNA1 (OsDmc1) from rice have been reported earlier. In the present study, DNA renaturation and strand exchange activities of OsDmc1 have been studied, in real time and without the steps of deproteinization, using fluorescence resonance energy transfer (FRET). The extent as well as the rate of renaturation is the highest in conditions that contain ATP, but significantly less when ATP is replaced by slowly hydrolysable analogues of ATP, namely adenosine 5'-(beta,gamma-imido) triphosphate (AMP-PNP) or adenosine 5'-O-(3-thio triphosphate) (ATP-gamma-S), where the former was substantially poorer than the latter in facilitating the renaturation function. FRET assay results also revealed OsDmc1 protein concentration dependent strand exchange function, where the activity was the fastest in the presence of ATP, whereas in the absence of a nucleotide cofactor it was several fold ( approximately 15-fold) slower. Interestingly, strand exchange, in reactions where ATP was replaced with AMP-PNP or ATP-gamma-S, was somewhat slower than that of even minus nucleotide cofactor control. Notwithstanding the slow rates, the reactions with no nucleotide cofactor or with ATP-analogues did reach the same steady state level as seen in ATP reaction. FRET changes were unaffected by the steps of deproteinization following OsDmc1 reaction, suggesting that the assay results reflected stable events involving exchanges of homologous DNA strands. All these results, put together, suggest that OsDmc1 catalyses homologous renaturation as well as strand exchange events where ATP hydrolysis seems to critically decide the rates of the reaction system. These studies open up new facets of a plant recombinase function in relation to the role of ATP hydrolysis. PMID- 16689936 TI - Integral membrane proteins in the mitochondrial outer membrane of Saccharomyces cerevisiae. AB - Mitochondria evolved from a bacterial endosymbiont ancestor in which the integral outer membrane proteins would have been beta-barrel structured within the plane of the membrane. Initial proteomics on the outer membrane from yeast mitochondria suggest that while most of the protein components are integral in the membrane, most of these mitochondrial proteins behave as if they have alpha-helical transmembrane domains, rather than beta-barrels. These proteins are usually predicted to have a single alpha-helical transmembrane segment at either the N- or C-terminus, however, more complex topologies are also seen. We purified the novel outer membrane protein Om14 and show it is encoded in the gene YBR230c. Protein sequencing revealed an intron is spliced from the transcript, and both transcription from the YBR230c gene and steady-state level of the Om14 protein is dramatically less in cells grown on glucose than in cells grown on nonfermentable carbon sources. Hydropathy predictions together with data from limited protease digestion show three alpha-helical transmembrane segments in Om14. The alpha helical outer membrane proteins provide functions derived after the endosymbiotic event, and require the translocase in the outer mitochondrial membrane complex for insertion into the outer membrane. PMID- 16689937 TI - Glucuronate, the precursor of vitamin C, is directly formed from UDP-glucuronate in liver. AB - The conversion of UDP-glucuronate to glucuronate, usually thought to proceed by way of glucuronate 1-phosphate, is a site for short-term regulation of vitamin C synthesis by metyrapone and other xenobiotics in isolated rat hepatocytes. Our purpose was to explore the mechanism of this effect in cell-free systems. Metyrapone and other xenobiotics stimulated, by approximately threefold, the formation of glucuronate from UDP-glucuronate in liver extracts enriched with ATP Mg, but did not affect the formation of glucuronate 1-phosphate from UDP glucuronate or the conversion of glucuronate 1-phosphate to glucuronate. This and other data indicated that glucuronate 1-phosphate is not an intermediate in glucuronate formation from UDP-glucuronate, suggesting that this reaction is catalysed by a 'UDP-glucuronidase'. UDP-glucuronidase was present mainly in the microsomal fraction, where its activity was stimulated by UDP-N acetylglucosamine, known to stimulate UDP-glucuronosyltransferases by enhancing the transport of UDP-glucuronate across the endoplasmic reticulum membrane. UDP glucuronidase and UDP-glucuronosyltransferases displayed similar sensitivities to various detergents, which stimulated at low concentrations and generally inhibited at higher concentrations. Substrates of glucuronidation inhibited UDP glucuronidase activity, suggesting that the latter is contributed by UDP glucuronosyltransferase(s). Inhibitors of beta-glucuronidase and esterases did not affect the formation of glucuronate, arguing against the involvement of a glucuronidation-deglucuronidation cycle. The sensitivity of UDP-glucuronidase to metyrapone and other stimulatory xenobiotics was lost in washed microsomes, even in the presence of ATP-Mg, but it could be restored by adding a heated liver high speed supernatant or CoASH. In conclusion, glucuronate formation in liver is catalysed by a UDP-glucuronidase which is closely related to UDP glucuronosyltransferases. Metyrapone and other xenobiotics stimulate UDP glucuronidase by antagonizing the inhibition exerted, presumably indirectly, by a combination of ATP-Mg and CoASH. PMID- 16689938 TI - Final steps in the catabolism of nicotine. AB - New enzymes of nicotine catabolism instrumental in the detoxification of the tobacco alkaloid by Arthrobacter nicotinovorans pAO1 have been identified and characterized. Nicotine breakdown leads to the formation of nicotine blue from the hydroxylated pyridine ring and of gamma-N-methylaminobutyrate (CH(3)-4 aminobutyrate) from the pyrrolidine ring of the molecule. Surprisingly, two alternative pathways for the final steps in the catabolism of CH(3)-4 aminobutyrate could be identified. CH(3)-4-aminobutyrate may be demethylated to gamma-N-aminobutyrate by the recently identified gamma-N-methylaminobutyrate oxidase. In an alternative pathway, an amine oxidase with noncovalently bound FAD and of novel substrate specificity removed methylamine from CH(3)-4-aminobutyrate with the formation of succinic semialdehyde. Succinic semialdehyde was converted to succinate by a NADP(+)-dependent succinic semialdehyde dehydrogenase. Succinate may enter the citric acid cycle completing the catabolism of the pyrrolidine moiety of nicotine. Expression of the genes of these enzymes was dependent on the presence of nicotine in the growth medium. Thus, two enzymes of the nicotine regulon, gamma-N-methylaminobutyrate oxidase and amine oxidase share the same substrate. The K(m) of 2.5 mM and k(cat) of 1230 s(-1) for amine oxidase vs. K(m) of 140 microM and k(cat) of 800 s(-1) for gamma-N-methylaminobutyrate oxidase, determined in vitro with the purified recombinant enzymes, may suggest that demethylation predominates over deamination of CH(3)-4-aminobutyrate. However, bacteria grown on [(14)C]nicotine secreted [(14)C]methylamine into the medium, indicating that the pathway to succinate is active in vivo. PMID- 16689939 TI - Transient silencing of Plasmodium falciparum bifunctional glucose-6-phosphate dehydrogenase- 6-phosphogluconolactonase. AB - The bifunctional enzyme glucose-6-phosphate dehydrogenase-6 phosphogluconolactonase (G6PD-6PGL) found in Plasmodium falciparum has unique structural and functional characteristics restricted to this genus. This study was designed to examine the effects of RNA-mediated PfG6PD-6PGL gene silencing in cultures of P. falciparum on the expression of parasite antioxidant defense genes at the transcription level. The highest degree of G6PD-6PGL silencing achieved was 86% at the mRNA level, with a recovery to almost normal levels within 24 h, indicating only transient diminished expression of the PfG6PD-6PGL gene. PfG6PD 6PGL silencing caused arrest of the trophozoite stage and enhanced gametocyte formation. In addition, an immediate transcriptional response was shown by thioredoxin reductase suggesting that P. falciparum G6PD-6PGL plays a physiological role in the specific response of the parasite to intracellullar oxidative stress. P. falciparum transfection with an empty DNA vector also promoted intracellular stress, as determined by mRNA up-regulation of antioxidant genes. Collectively, our findings point to an important role for this enzyme in the parasite's infection cycle. The different characteristics of G6PD-6PGL with respect to its homologue in the host make it an ideal target for therapeutic strategies. PMID- 16689940 TI - The most C-terminal tri-glycine segment within the polyglycine stretch of the pea Toc75 transit peptide plays a critical role for targeting the protein to the chloroplast outer envelope membrane. AB - The protein translocation channel at the outer envelope membrane of chloroplasts (Toc75) is synthesized as a larger precursor with an N-terminal transit peptide. Within the transit peptide of the pea Toc75, a major portion of the 10 amino acid long stretch that contains nine glycine residues was shown to be necessary for directing the protein to the chloroplast outer membrane in vitro. In order to get insights into the mechanism by which the polyglycine stretch mediates correct targeting, we divided it into three tri-glycine segments and examined the importance of each domain in targeting specificity in vitro. Replacement of the most C-terminal segment with alanine residues resulted in mistargeting the protein to the stroma, while exchange of either of the other two tri-glycine regions had no effect on correct targeting. Furthermore, simultaneous replacement of the N-terminal and middle tri-glycine segments with alanine repeats did not cause mistargeting of the protein as much as those of the N- and C-terminal, or the middle and C-terminal segments. These results indicate that the most C terminal tri-glycine segment is important for correct targeting. Exchanging this portion with a repeat of leucine or glutamic acid also caused missorting of Toc75 to the stroma. By contrast, its replacement with repeats of asparagine, aspartic acid, serine, and proline did not largely affect correct targeting. These data suggest that relatively compact and nonhydrophobic side chains in this particular region play a crucial role in correct sorting of Toc75. PMID- 16689941 TI - A second independent resistance mechanism to Bacillus sphaericus binary toxin targets its alpha-glucosidase receptor in Culex quinquefasciatus. AB - The entomopathogen Bacillus sphaericus is an important tool for the vector control of Culex sp., and its effectiveness has been validated in field trials. The appearance of resistance to this bacterium, however, remains a threat to its use, and attempts have been made to understand the resistance mechanisms. Previous work showed that the resistance to B. sphaericus in a Culex quinquefasciatus colony is associated with the absence of the approximately 60 kDa binary toxin receptor in larvae midgut microvilli. Here, the gene encoding the C. quinquefasciatus toxin receptor, Cqm1, was cloned and sequenced from a susceptible colony. The deduced amino-acid sequence confirmed its identity as an alpha-glucosidase, and analysis of the corresponding gene sequence from resistant larvae implicated a 19-nucleotide deletion as the basis for resistance. This deletion changes the ORF and originates a premature stop codon, which prevents the synthesis of the full-length Cqm1. Expression of the truncated protein, however, was not detected when whole larvae extracts were probed with antibodies raised against an N-terminal 45-kDa recombinant fragment of Cqm1. It seems that the premature stop codon directs the mutated cqm1 to the nonsense-mediated decay pathway of mRNA degradation. In-gel assays confirmed that a single alpha glucosidase protein is missing from the resistant colony. Further in vitro affinity assays showed that the recombinant fragment binds to the toxin, and mapped the binding site to the N-terminus of the receptor. PMID- 16689942 TI - Identification of GAS-dependent interferon-sensitive target genes whose transcription is STAT2-dependent but ISGF3-independent. AB - Signal transducer and activator of transcription 2 (STAT2) is best known as a critical transactivator component of the interferon-stimulated gene factor 3 (ISGF3) complex that drives the expression of many interferon (IFN)-inducible genes. However, STAT2 is also involved in DNA binding in non-ISGF3 transcriptional complexes. We used a DNA microarray to survey the expression of genes regulated by IFN-inducible, STAT2-dependent DNA binding, and compared the cDNAs of IFN-treated cells overexpressing intact STAT2 to those of IFN-treated cells overexpressing mutated STAT2 lacking the DNA binding domain. The IFN inducible expression of genes known to be regulated by ISGF3 was similar in both cases. However, a subset of IFN-inducible genes was identified whose expression was decreased in cells expressing the mutated STAT2. Importantly, these genes all contained gamma-activated sequence (GAS)-like elements in their 5' flanking sequences. Our data reveal the existence of a collection of GAS-regulated target genes whose expression is IFN-inducible and independent of ISGF3 but highly dependent on the STAT2 DNA binding domain. This report is the first analysis of the contribution of the STAT2 DNA binding domain to IFN responses on a global basis, and shows that STAT2 is required for the IFN-inducible activation of the full spectrum of GAS target genes. PMID- 16689943 TI - Novel dissociation mechanism of a polychaetous annelid extracellular haemoglobin. AB - The extracellular haemoglobin of the marine polychaete, Arenicola marina, is a hexagonal bilayer haemoglobin of approximately 3600 kDa, formed by the covalent and noncovalent association of many copies of both globin subunits (monomer and trimer) and nonglobin or 'linker' subunits. In order to analyse the interactions between globin and linker subunits, dissociation and reassociation experiments were carried out under whereby Arenicola hexagonal bilayer haemoglobin was exposed to urea and alkaline pH and the effect was followed by gel filtration, SDS/PAGE, UV-visible spectrophotometry, electrospray-ionization MS, multiangle laser light scattering and transmission electron microscopy. The analysis of Arenicola haemoglobin dissociation indicates a novel and complex mechanism of dissociation compared with other annelid extracellular haemoglobins studied to date. Even though the chemically induced dissociation triggers partial degradation of some subunits, spontaneous reassociation was observed, to some extent. Parallel dissociation of Lumbricus haemoglobin under similar conditions shows striking differences that allow us to propose a hypothesis on the nature of the intersubunit contacts that are essential to form and to hold such a complex quaternary structure. PMID- 16689945 TI - Condition-dependent nesting in bluegill sunfish Lepomis macrochirus. AB - 1. There are two prominent, nonmutually exclusive hypotheses to explain the timing of reproduction in animals: energetic constraint and adaptive behaviour. 2. We tested these hypotheses by quantifying the costs and benefits of nesting at different times in the season for male bluegill sunfish Lepomis macrochirus, a species with paternal care, in Lake Opinicon (Ontario, Canada). 3. The value of nesting at different times during the breeding season (RSb) was determined from spawning individuals as RSb = Pb x Cb x Sb x Ob, where Pb is the probability of spawning during each bout b, Cb is the expected brood size, Sb is the expected brood survivorship to 'swim-up', and Ob is the survivorship of free-swimming fry to age 1 year. 4. The results show that the value of nesting peaks during the middle of the season. However, nesting patterns varied with male condition and not all males nested at the peak. 5. Larger males, which were able to nest multiple times, first nested early in the season when overwinter survivorship of offspring and renesting opportunities later in the season were maximized. These males had the highest seasonal reproductive success. 6. Smaller males, which nested a single time, delayed nesting until the middle of the season when spawning opportunities and brood sizes were greatest. 7. These data suggest that both energetics and adaptive behaviour play roles in determining the timing of reproduction. PMID- 16689946 TI - Incorporating movement into models of grey seal population dynamics. AB - 1. One of the most difficult problems in developing spatially explicit models of population dynamics is the validation and parameterization of the movement process. We show how movement models derived from capture-recapture analysis can be improved by incorporating them into a spatially explicit metapopulation model that is fitted to a time series of abundance data. 2. We applied multisite capture-recapture analysis techniques to photo-identification data collected from female grey seals at the four main breeding colonies in the North Sea between 1999 and 2001. The best-fitting movement models were then incorporated into state space metapopulation models that explicitly accounted for demographic and observational stochasticity. 3. These metapopulation models were fitted to a 20 year time series of pup production data for each colony using a Bayesian approach. The best-fitting model, based on the Akaike Information Criterion (AIC), had only a single movement parameter, whose confidence interval was 82% less than that obtained from the capture-recapture study, but there was some support for a model that included an effect of distance between colonies. 4. The state-space modelling provided improved estimates of other demographic parameters. 5. The incorporation of movement, and the way in which it was modelled, affected both local and regional dynamics. These differences were most evident as colonies approached their carrying capacities, suggesting that our ability to discriminate between models should improve as the length of the grey seal time series increases. PMID- 16689947 TI - Abundance, spatial variance and occupancy: arthropod species distribution in the Azores. AB - 1. The positive abundance-occupancy and abundance-variance relationships are two of the most widely documented patterns in population and community ecology. 2. Recently, a general model has been proposed linking the mean abundance, the spatial variance in abundance, and the occupancy of species. A striking feature of this model is that it consists explicitly of the three variables abundance, variance and occupancy, and no extra parameters are involved. However, little is known about how well the model performs. 3. Here, we show that the abundance variance-occupancy model fits extremely well to data on the abundance, variance and occupancy of a large number of arthropod species in natural forest patches in the Azores, at three spatial extents, and distinguishing between species of different colonization status. Indeed, virtually all variation about the bivariate abundance-occupancy and abundance-variance relationships is effectively explained by the third missing variable (variance in abundance in the case of the abundance-occupancy relationship, and occupancy in the case of the abundance variance relationship). 4. Introduced species tend to exhibit lower densities, less spatial variance in these densities, and occupy fewer sites than native and endemic species. None the less, they all lie on the same bivariate abundance occupancy and abundance-variance, and trivariate abundance-variance-occupancy, relationships. 5. Density, spatial variance in density, and occupancy appear to be all the things one needs to know to describe much of the spatial distribution of species. PMID- 16689948 TI - Rapidly advancing laying date in a seabird and the changing advantage of early reproduction. AB - 1. Bird ringing schemes have collected immense amounts of data on timing of breeding for over 100 years. These data provide an unexploited source of information on temporal change in breeding date. 2. We investigated changes in breeding date of the Arctic tern Sterna paradisaea Pont. in Denmark during 1929 98, using information on ringing date of young. 3. Mean ringing date advanced by over 18 days during 70 years, while there was no temporal change in variance in date. 4. Advanced mean ringing date was explained by an increase in mean temperature during April and May and an increase in North Atlantic Oscillation (NAO) index for May. 5. Variance in ringing date increased in years with high temperatures in April and high NAO index values in April. 6. There was changing temporal patterns of selection for early breeding as reflected by analyses of the difference in mean ringing date for Arctic tern young that were subsequently recorded as survivors and mean ringing date for all young. The intensity of selection on breeding date changed from favouring late breeding in the 1930s to favouring early breeding during the 1990s. 7. Analyses of bird ringing information for millions of offspring of hundreds of bird species deposited in national ringing schemes may provide unlimited access to long-term time series of reproductive variables. PMID- 16689949 TI - Validation of a randomization procedure to assess animal habitat preferences: microhabitat use of tiger sharks in a seagrass ecosystem. AB - 1. Tiger sharks Galeocerdo cuvier are important predators in a variety of nearshore communities, including the seagrass ecosystem of Shark Bay, Western Australia. Because tiger sharks are known to influence spatial distributions of multiple prey species, it is important to understand how they use habitats at a variety of spatial scales. We used a combination of catch rates and acoustic tracking to determine tiger shark microhabitat use in Shark Bay. 2. Comparing habitat-use data from tracking against the null hypothesis of no habitat preference is hindered in Shark Bay, as elsewhere, by the difficulty of defining expected habitat use given random movement. We used randomization procedures to generate expected habitat use in the absence of habitat preference and expected habitat use differences among groups (e.g. males and females). We tested the performance of these protocols using simulated data sets with known habitat preferences. 3. The technique correctly classified sets of simulated tracks as displaying a preference or not and was a conservative test for differences in habitat preferences between subgroups of tracks (e.g. males vs. females). 4. Sharks preferred shallow habitats over deep ones, and preferred shallow edge microhabitats over shallow interior ones. The use of shallow edges likely increases encounter rates with potential prey and may have profound consequences for the dynamics of Shark Bay's seagrass ecosystem through indirect effects transmitted by grazers that are common prey of tiger sharks. 5. Females showed a greater tendency to use shallow edge microhabitats than did males; this pattern was not detected by traditional analysis techniques. 6. The randomization procedures presented here are applicable to many field studies that use tracking by allowing researchers both to determine overall habitat preferences and to identify differences in habitat use between groups within their sample. PMID- 16689950 TI - Effects of food availability on temporal activity patterns and growth of Atlantic salmon. AB - 1. Patterns of sheltering and activity are of fundamental importance in the ecology of animals and in determining interactions among predators and prey. Balancing decreased mortality risk when sheltering with increased feeding rate when exposed is believed to be a key determinant of diel patterns of sheltering in many animals. 2. Despite lower foraging efficiency at night than during the day, Atlantic salmon Salmo salar parr are nocturnal during winter and at low summer temperatures. Nocturnal activity also occurs at warm water temperatures during summer, but little is known about the functional significance of this behaviour. 3. This study aimed to determine: (1) the preferred activity and shelter pattern of Atlantic salmon parr during warm summer months, and (2) their response to variations in food availability when balancing growth rate (G) and mortality risk (M), as expressed through time out of shelter. We differentiated among four potential responses to reduced food availability: (1) no response; (2) G decreases but M remains constant; (3) G remains constant but M increases; and (4) G decreases and M increases. 4. Time exposed from shelter was inversely related to food availability. Fish subject to high food availability were significantly less active during the day than those with restricted rations. However, food availability had no significant effect on the extent to which fish were active at night. There was no evidence of variation in growth rate with food availability. 5. Salmon were predominantly nocturnal at high ration levels, consistent with their previously reported behaviour during winter. Rather than switching to diurnal behaviour at high temperatures per se, as previously was supposed, it appears that the fish are diurnal only to the extent needed to sustain a growth rate, and this extent depends on food availability. 6. Atlantic salmon parr modulate the amount of time they are active rather than growth when responding to variations in food availability over an order of magnitude. PMID- 16689951 TI - Short- and long-term consequences of thermal variation in the larval environment of anurans. AB - 1. To survive adverse or unpredictable conditions in the ontogenetic environment, many organisms retain a level of phenotypic plasticity that allows them to meet the challenges of rapidly changing conditions. Larval anurans are widely known for their ability to modify behaviour, morphology and physiological processes during development, making them an ideal model system for studies of environmental effects on phenotypic traits. Although temperature is one of the most important factors influencing the growth, development and metamorphic condition of larval anurans, many studies have failed to include ecologically relevant thermal fluctuations among their treatments. 2. We compared the growth and age at metamorphosis of striped marsh frogs Limnodynastes peronii raised in a diurnally fluctuating thermal regime and a stable regime of the same mean temperature. We then assessed the long-term effects of the larval environment on the morphology and performance of post-metamorphic frogs. 3. Larval L. peronii from the fluctuating treatment were significantly longer throughout development and metamorphosed about 5 days earlier. Frogs from the fluctuating group metamorphosed at a smaller mass and in poorer condition compared with the stable group, and had proportionally shorter legs. 4. Frogs from the fluctuating group showed greater jumping performance at metamorphosis and less degradation in performance during a 10-week dormancy. Treatment differences in performance could not be explained by whole-animal morphological variation, suggesting improved contractile properties of the muscles in the fluctuating group. PMID- 16689952 TI - Does life-history variability in salmonids affect habitat use by juveniles? A comparison among streams open and closed to anadromy. AB - 1. Migratory and resident forms of salmonids coexist in many river systems. Although such coexistence is widespread, little is known about its ecological basis and no studies have compared the habitat use of premigratory juveniles and residents. 2. We employed a comparative approach to explore the differential habitat use of juvenile anadromous and resident brook trout. This required the investigation of habitat use in streams closed to anadromy, containing only resident brook trout Salvelinus fontinalis ('resident-only' streams) and streams open to anadromy, containing coexisting Atlantic salmon Salmo salar and anadromous and resident brook trout ('migrant-resident' streams). 3. We demonstrate that fast habitats (riffles) are occupied more frequently in streams with migratory brook trout relative to riffle habitats of streams with only resident brook trout. In contrast, occupation of slow current velocities (pools) was observed in both migrant-resident and resident-only streams as both stream types contain resident brook trout. The net effect is a wider distribution of occupied habitats (pool and riffles) in migrant-resident streams relative to resident-only streams, resulting in few, if any, unused habitats. 4. These results are consistent with previously reported bioenergetic, morphological and stable isotope differences observed between anadromous and resident brook trout. 5. Our findings suggest that a link exists between juvenile habitat use, metabolic costs and life-history strategies. PMID- 16689953 TI - Flexible architecture of inducible morphological plasticity. AB - 1. Predator-induced morphological defences are produced in response to an emergent predator regime. In natural systems, prey organisms usually experience temporal shifting of the composition of the predator assemblage and of the intensity of predation risk from each predator species. Although, a repetitive morphological change in response to a sequential shift of the predator regime such as alteration of the predator species or diminution of the predation risk may be adaptive, such flexible inducible morphological defences are not ubiquitous. 2. We experimentally addressed whether a flexible inducible morphological defence is accomplished in response to serial changes in the predation regime, using a model prey species which adopt different defensive morphological phenotypes in response to different predator species. Rana pirica (Matsui) tadpoles increased body depth and tail depth against the predatory larval salamander Hynobius retardatus (Dunn); on the other hand, they only increased tail depth against the predatory larval dragonfly Aeshna nigroflava (Martin). 3. Rana pirica tadpoles with the predator-specific phenotypes were subjected to removal or exchange of the predator species. After removal of the predator species, tadpoles with each predator-specific phenotype changed their phenotype to the nondefensive basic one, suggesting that both predator-specific phenotypes are costly to maintain. After an exchange of the predator species, tadpoles with each predator-specific phenotype reciprocally, flexibly shifted their phenotype to the now more suitable predator-specific one only by modifying their body part. The partial modification can effectively reduce time and energy expenditures involved in repetitive morphological changes, and therefore suggest that the costs of the flexible morphological changes are reduced. PMID- 16689954 TI - Behaviourally mediated indirect effects: interference competition increases predation mortality in foraging redshanks. AB - 1. The effect of competition for a limiting resource on the population dynamics of competitors is usually assumed to operate directly through starvation, yet may also affect survival indirectly through behaviourally mediated effects that affect risk of predation. Thus, competition can affect more than two trophic levels, and we aim here to provide an example of this. 2. We show that the foraging success of redshanks Tringa totanus (L.) foraging on active prey was highest in the front of flocks, whereas this was not the case for redshanks foraging on inactive prey. Also, when foraging on active prey, foraging success in a flock decreased as more birds passed through a patch, while overall foraging success was not lower on subsequent visits to the same patch. Thus, redshanks foraging on active prey suffered from interference competition, whereas this was not the case for redshanks foraging on inactive prey. 3. This interference competition led to differences in activity: redshanks attaining a lower foraging success had a higher walking rate. Greater activity was associated with wider flock spacing and shorter distances to cover, which has previously been shown to increase predation risk and mortality from sparrowhawks Accipiter nisus (L.). 4. We conclude that behavioural adaptations of prey species can lead to interference competition in foraging redshanks, and thus can affect their predation risk and mortality through increased activity. This study is one of the first to show how interference competition can be a mechanism for behaviourally mediated indirect effects, and provides further evidence for the suggestion that a single species occupying an intermediate trophic level may be simultaneously top-down controlled by a predator and bottom-up controlled by a behavioural response of its prey. PMID- 16689955 TI - Influence of maternal mass and condition on energy transfer in Weddell seals. AB - 1. Environmental variation influences food abundance and availability, which is reflected in the reproductive success of top predators. We examined maternal expenditure, offspring mass and condition for Weddell seals in 2 years when individuals exhibited marked differences in these traits. 2. For females weighing > or = 355 kg there was a positive relationship between maternal post-partum mass (MPPM) and lactation length, but below this there was no relationship, suggesting that heavier females were able to increase lactation length but lighter females were restricted to a minimum lactation period of 33 days. 3. Overall, females were heavier in 2002, but in 2003 shorter females were lighter than similar-sized females in 2002 suggesting that the effects of environmental variability on foraging success and condition are more pronounced in smaller individuals. 4. There was no relationship between MPPM and pup birth mass, indicating pre-partum investment did not differ between years. However, there was a positive relationship between MPPM and pup mass gain. Mass and energy transfer efficiency were 10.2 and 5.4% higher in 2002 than 2003, which suggests costs associated with a putatively poor-resource year were delayed until lactation. 5. Heavier females lost a higher proportion of mass during lactation in both years, so smaller females may not have been able to provide more to their offspring to wean a pup of similar size to larger females. 6. MPPM had only a small influence on total body lipid; therefore, regardless of mass, females had the same relative body composition. Females with male pups lost a higher percentage of lipid than those with female pups, but by the end of lactation female pups had 4.5% higher lipid content than males. 7. It appears that for Weddell seals the consequences of environmentally induced variation in food availability are manifested in differences in maternal mass and expenditure during lactation. These differences translate to changes in pup mass and condition at weaning with potential consequences for future survival and recruitment. PMID- 16689956 TI - Pattern and process in the ecological biogeography of European freshwater fish. AB - 1. Species lists for regions of Europe defined by Illies (1978, Limnofauna Europaea, 2nd edn. Gustav Fischer Verlag, Stuttgart), and augmented by information from Maitland (2000, Guide to Freshwater Fish of Britain and Europe. Hamlyn, London), are used to describe patterns in freshwater fish species richness and to examine the contribution of habitat preference, migration, body size and glacial history to these patterns. 2. The number of non-endemic species declines to the north and west, with increasing distance from the Ponto-Caspian region, the main source area, whereas endemic species richness declines only with latitude. 3. Habitat generalists tend to be migratory while riverine specialists are usually resident. Similar numbers of riverine species and generalists occur in Europe as a whole but generalists dominate in regional faunas and, to an increasing extent, in more isolated, formerly glaciated areas. Very few lacustrine specialists were found, reflecting the geologically ephemeral nature of lakes. Only 8% of riverine species have colonized glaciated areas, compared with more than half the generalist species, and the number declines rapidly with increasing distance from the source area. 4. Diadromous species show no geographical variation in species richness but potamodromous and resident species are affected by glaciation and by mountain and marine barriers. 5. The mean body size of regional faunas increases with latitude because there are relatively fewer small species in more distant, glaciated areas. 6. About half the species occurring in Europe are restricted to one region and the majority of these endemics occur in barrier regions with Mediterranean climates. Species in glaciated regions have much larger range sizes. Habitat preference and migration type, not body size, are the main determinants of range size. 7. Freshwater habitat availability varies across Europe with glaciated areas having more lakes of a given size than unglaciated areas. Catchment size is greatest at mid latitudes. For a given catchment size rivers in glaciated areas are shorter. 8. The results support the notion that habitat variability, on both short and long time scales, favours colonization ability, which requires large body size. 9. As a result of their limited vagility northern fish faunas are depauperate, show high levels of plasticity and polymorphism and may show elevated speciation rates. The isolated southern faunas of the Iberian and Italian peninsulas and the Balkans are rich in endemic species but may be subject to extinctions because of the spread of the highly seasonal Mediterranean climate. PMID- 16689957 TI - On plotting species abundance distributions. AB - 1. There has been a revival of interest in species abundance distribution (SAD) models, stimulated by the claim that the log-normal distribution gave an underestimate of the observed numbers of rare species in species-rich assemblages. This led to the development of the neutral Zero Sum Multinomial distribution (ZSM) to better fit the observed data. 2. Yet plots of SADs, purportedly of the same data, showed differences in frequencies of species and of statistical fits to the ZSM and log-normal models due to the use of different binning methods. 3. We plot six different binning methods for the Barro Colorado Island (BCI) tropical tree data. The appearances of the curves are very different for the different binning methods. Consequently, the fits to different models may vary depending on the binning system used. 4. There is no agreed binning method for SAD plots. Our analysis suggests that a simple doubling of the number of individuals per species in each bin is perhaps the most practical one for illustrative purposes. Alternatively rank-abundance plots should be used. 5. For fitting and testing models exact methods have been developed and application of these does not require binning of data. Errors are introduced unnecessarily if data are binned before testing goodness-of-fit to models. PMID- 16689958 TI - Urban amphibian assemblages as metacommunities. AB - 1. Urban ecosystems are expanding throughout the world, and urban ecology is attracting increasing research interest. Some authors have questioned the value of existing ecological theories for understanding the processes and consequences of urbanization. 2. In order to assess the applicability of metacommunity theory to urban systems, I evaluated three assumptions that underlie the theory - the effect of patch area, the effect of patch isolation, and species-environment relations - using data on assemblages of pond-breeding amphibians in the Greater Melbourne area of Australia. I also assessed the relative impact of habitat fragmentation, habitat isolation, and changes to habitat quality on these assemblages. 3. Poisson regression modelling provided support for an important increase in species richness with patch area (pond size) and a decrease in species richness with increasing patch isolation, as measured by surrounding road cover. Holding all other variables constant, species richness was predicted to be 2.8-5.5 times higher at the largest pond than at the smallest, while the most isolated pond was predicted to have 12-19% of the species richness of the least isolated pond. Thus, the data were consistent with the first two assumptions of metacommunity theory evaluated. 4. The quality of habitat at a pond was also important, with a predicted 44-56% decrease in the number of species detected at ponds with a surrounding vertical wall compared with those with a gently sloping bank. This demonstrates that environmental differences between habitat patches were also influencing amphibian assemblages, providing support for the species sorting and/or mass-effect perspectives of metacommunity theory. 5. Without management intervention, urbanization may lead to a reduction in the number of amphibian species persisting in urban ponds, particularly where increasing isolation of ponds by roads and associated infrastructure reduces the probability of re-colonization following local extinction. Journal of Animal Ecology (2006) 75, 757-764 doi: 10.1111/j.1365-2656.2006.01096.x. PMID- 16689959 TI - Competing species in a changing climate: effects of recruitment disturbances on two interacting barnacle species. AB - 1. The climate is changing and data-based simulation models can be a valuable tool for predicting population response to such changes and investigate the mechanisms of population change. In this study, a data-based two-species matrix model was constructed to explore the possible effects of elevated sea surface temperature (i.e. climate change) on the interaction between open populations of the south Atlantic barnacle species Chthamalus montagui and the boreal species Semibalanus balanoides in the north-east Atlantic. 2. First, the model was used to perform an elasticity analysis to determine the relative importance of recruitment and survival in the interaction. Further, three scenarios of changes in recruitment, related to climate change, were investigated with model simulations: (i) increased frequencies of low recruitment for S. balanoides; (ii) increased frequencies of high recruitment for C. montagui; (iii) a combination of (i) and (ii). 3. Model simulations showed that in present environmental conditions, S. balanoides occupied most of the space and dominated the interaction through high recruitment and survival. These results matched independent field observations, which validated the model for further analyses. 4. The elasticity analyses showed that although free space was available there was competition for space during recruitment intervals. It was also shown that both populations were sensitive to changes in recruitment. 5. Introducing the three scenarios of recruitment disturbances led to large changes in species abundance and free space. The most significant changes were found when scenario (i) and (ii) were combined, producing a shift in species dynamics towards C. montagui dominance. This demonstrates that recruitment can be an important mechanism in the interaction between populations and that the population response to changes in recruitment depends on the added response of interacting species. 6. In a more general context, this model shows that increased sea surface temperature could rapidly lead to increased competition from southern species at higher latitudes. This might accelerate the effects of climate change on the species distribution at these latitudes and eventually lead to changes in community dynamics on temperate and subarctic shores. PMID- 16689960 TI - Modelling the impacts of two exotic invasive species on a native butterfly: top down vs. bottom-up effects. AB - 1. Exotic invasive species can influence population dynamics of native species through top-down or bottom-up forces. The present study examined separate and interactive effects of multiple exotic species invasions on the native mustard white butterfly, Pieris napi oleracea Harris (Lepidoptera: Pieridae), using a stochastic simulation model. 2. P. n. oleracea populations in North America have decreased regionally since the 1860s. Competition with an exotic congener (P. rapae L.), loss of native host plants and parasitism by the introduced broconid wasp (Cotesia glomerata L.), have been suggested to be independently responsible for its decline. The present study examined these hypotheses, as well as an alternative, invasion by an exotic crucifer, garlic mustard (Alliaria petiolata[Bieb.] Cavara & Grande). 3. A stochastic simulation model of P. n. oleracea population dynamics revealed that decreasing the number of host plants available for oviposition and larval development (i.e. habitat loss), sharply reduced the probability of populations persistence and decreased population size for those that persisted. 4. Simulated invasion by garlic mustard also substantially decreased both probability of persistence (= 0 at approximately 50% cover) and mean population size. Persistence probability never reached zero under any C. glomerata scenarios, even when larval mortality in the second generation due to parasitism was 100%. The impact of garlic mustard was intensified by the addition of C. glomerata parasitism. 5. Results suggest that bottom-up forces, loss of host plants through forest understorey loss and/or garlic mustard invasion are the most important forces driving P. n. oleracea population decline. Parasitism by C. glomerata may interact to reduce P. n. oleracea populations more rapidly, but appears insufficient alone to cause local extinction. PMID- 16689961 TI - Consequences for host-parasitoid interactions of grazing-dependent habitat heterogeneity. AB - 1. Environmental heterogeneity can produce effects that cascade up to higher trophic levels and affect species interactions. We hypothesized that grazing dependent habitat heterogeneity and grazing-independent host plant heterogeneity would influence directly and indirectly a host-parasitoid interaction in a woodland habitat. 2. Thistles were planted randomly in 20 birch woodlands, half of which are grazed by cattle. The abundances of two species of seed herbivore and their shared parasitoid were measured, and related to habitat and host-plant heterogeneity. 3. The presence of cattle grazing created a structurally and compositionally distinct plant assemblage from the ungrazed seminatural situation. Grazing did not affect the number or dispersion of the host plant underpinning the host-parasitoid interaction. 4. The density of one insect herbivore, Tephritis conura, and its parasitoid Pteromalus elevatus was significantly increased by the presence of cattle; but another herbivore, Xyphosia miliaria, was unaffected. The percentage of parasitism of T. conura was increased in grazed habitat occurring at twice the rate found in ungrazed habitat. 5. The increase in T. conura abundance was correlated with increased species richness and cover of forbs in grazed sites. This effect of grazing dependent habitat variation on host insect density cascaded up to parasitoid density and percentage of parasitism. Habitat heterogeneity had a further direct, positive effect on parasitoid density and percentage of parasitism after controlling for host-insect density. 6. Independent of grazing, heterogeneity in host-plant flowering, architecture and stature further affected T. conura and its parasitoid's densities. Parasitoid density was also affected by the dispersion of the host plant. 7. A combination of habitat and host-plant scale environmental heterogeneity influenced a host-parasitoid interaction indirectly and directly, providing a rare example of an anthropogenic disturbance positively affecting a tertiary trophic level. This finding highlights the need to consider not only the importance of bottom-up effects for top-down processes, but also the role of environmental heterogeneity arising from anthropogenic disturbance for trophic interactions such as parasitism. PMID- 16689962 TI - Active and passive dispersal of an invading land snail in Mediterranean France. AB - 1. Land snail dispersal abilities are considered poor; however, the current invasion of the French Mediterranean region by Xeropicta derbentina (Krynicki 1836), as well as the past invasions of this region by several other species, seems to contradict this view. 2. Using a multilevel approach, from individual experimentation to landscape analysis, the dispersal abilities and mechanisms allowing the passive dispersal of X. derbentina are studied. 3. The colonization of Provence occurred by stratified diffusion, where short-range active dispersal occurs side by side with long-range passive dispersal. 4. Active dispersal is not as limited as previously thought. In the field, the capture-mark-recapture method recorded a maximum distance covered of 42 m in 6 months within a radius of 38 m from the original release point. 5. Temperature and humidity, and therefore the time of year, influence the main type of dispersal. Dispersal is active during wet periods and essentially passive in dry and hot months. 6. Heat avoidance behaviour is one of the mechanisms allowing passive dispersal. 7. Passive dispersal via human activities is the main determinant of X. derbentina distribution within the landscape. In comparison to other species, X. derbentina is found more often in the vicinity of a communication route. 8. These results show that land snails can cover large distances in a lifetime. The potential for active and passive dispersal described in this paper enables X. derbentina to be a successful invasive species and explains the rapid spread and current distribution of this species. PMID- 16689963 TI - Deep-diving foraging behaviour of sperm whales (Physeter macrocephalus). AB - 1. Digital tags were used to describe diving and vocal behaviour of sperm whales during 198 complete and partial foraging dives made by 37 individual sperm whales in the Atlantic Ocean, the Gulf of Mexico and the Ligurian Sea. 2. The maximum depth of dive averaged by individual differed across the three regions and was 985 m (SD = 124.3), 644 m (123.4) and 827 m (60.3), respectively. An average dive cycle consisted of a 45 min (6.3) dive with a 9 min (3.0) surface interval, with no significant differences among regions. On average, whales spent greater than 72% of their time in foraging dive cycles. 3. Whales produced regular clicks for 81% (4.1) of a dive and 64% (14.6) of the descent phase. The occurrence of buzz vocalizations (also called 'creaks') as an indicator of the foraging phase of a dive showed no difference in mean prey capture attempts per dive between regions [18 buzzes/dive (7.6)]. Sperm whales descended a mean of 392 m (144) from the start of regular clicking to the first buzz, which supports the hypothesis that regular clicks function as a long-range biosonar. 4. There were no significant differences in the duration of the foraging phase [28 min (6.0)] or percentage of the dive duration in the foraging phase [62% (7.3)] between the three regions, with an overall average proportion of time spent actively encountering prey during dive cycles of 0.53 (0.05). Whales maintained their time in the foraging phase by decreasing transit time for deeper foraging dives. 5. Similarity in foraging behaviour in the three regions and high diving efficiencies suggest that the success of sperm whales as mesopelagic predators is due in part to long-range echolocation of deep prey patches, efficient locomotion and a large aerobic capacity during diving. PMID- 16689964 TI - Climate change may account for the decline in British ring ouzels Turdus torquatus. AB - 1. Climate change is already affecting biodiversity, but the number of species for which reliable models relate weather and climate to demographic parameters is low. 2. We modelled the effect of temperature and rainfall on the breeding success and territory occupancy of ring ouzels Turdus torquatus (L.) in northern Britain, using data from a range of study areas, including one where there was a long-term decline in ring ouzel abundance. 3. Timing of breeding was significantly related to meteorological variables affecting birds in the early spring, though there was no evidence that laying dates had advanced. Breeding success was not significantly related to weather variables; instead, over 90% of annual variation in this parameter could be explained by density dependence. 4. Annual change in territory occupancy was linked to rainfall and temperature the preceding summer, after the main breeding season and to rainfall in the wintering grounds 24 months previously, coincident with the period of juniper Juniperus sp. (L.) flowering. High temperature in late summer, intermediate levels of late summer rainfall, and high spring rainfall in Morocco 24 months previously all had negative impacts on territory occupancy the following year. 5. All three weather variables have changed over recent decades, with a significant increase in summer temperature, a significant decrease in summer rainfall, and a nonsignificant decline in Moroccan spring rainfall. A model based on these trends alone predicted an annual decline in occupancy of 3.6% (compared with an observed decline of 1.2%), and suggested that increased summer temperatures may underlie declines in the British ring ouzel population. 6. Changes in summer temperature after the main breeding period could affect the survival rates of adult and/or juvenile birds. An improved understanding of the post-breeding ecology of ring ouzels is required to elucidate the mechanisms and causes of this relationship. Such knowledge might allow management aimed at buffering the impacts of climate change on ring ouzels. PMID- 16689966 TI - Nephrogenic systemic fibrosis: an emerging threat among renal patients. AB - Nephrogenic systemic fibrosis (NSF), also known as nephrogenic fibrosing dermopathy, is a scleroderma-like disease of the skin observed in patients with renal insufficiency. Since its recognition in 1997, relatively little has been published in the nephrology literature, despite indications that the disorder is becoming increasingly recognized in the United States and abroad. As nephrology professionals may be the first to encounter newly developing cases of NSF, it becomes imperative that they be able to recognize the symptoms and comorbidities and initiate tests and treatment. To date, the cause of NSF has not been established. Recent observations indicate that specific cells normally involved in wound repair and tissue remodeling may be aberrantly recruited to the skin and soft tissues of NSF patients. PMID- 16689967 TI - Lanthanum: a safe phosphate binder. AB - Accumulation of inorganic phosphate due to renal functional impairment contributes to the increased cardiovascular mortality observed in dialysis patients. Phosphate plays a causative role in the development of vascular calcification in renal failure; treatment with calcium-based phosphate binders and vitamin D can further increase the Ca x PO(4) product and add to the risk of ectopic mineralization. The new generation of calcium-free phosphate binders, sevelamer and lanthanum, can control hyperphosphatemia without adding to the patients calcium load. In this article, the metabolism of lanthanum carbonate and its effects in bone, liver and brain are discussed. Although lanthanum is a metal cation its effects are not comparable to those of aluminum. Indeed, in clinical studies no toxic effects of lanthanum have been reported after up to four years of follow-up. The bioavailability of lanthanum is extremely low. The effects observed in bone are due to phosphate depletion, with no signs of direct bone toxicity yet observed in rats or humans. The liver is the main route of excretion for lanthanum carbonate, which can be localized in the lysosomes of hepatocytes. No lanthanum could be detected in brain tissue. PMID- 16689968 TI - What should nephrologists do to maximize the use of arteriovenous fistulas? PMID- 16689969 TI - What should nephrologists do to maximize the use of arteriovenous fistulas? PMID- 16689970 TI - What should nephrologists do to maximize the use of arteriovenous fistulas? PMID- 16689971 TI - What should nephrologists do to maximize the use of arteriovenous fistulas? PMID- 16689972 TI - Diagnosis and management of sleep apnea syndrome and restless legs syndrome in dialysis patients. AB - Sleep complaints are very common in patients with end-stage renal disease (ESRD) and contribute to their impaired quality of life. Both obstructive and central sleep apnea syndromes are reported more often in patients on dialysis than in the general population. Impaired daytime functioning, sleepiness, and fatigue, as well as cognitive problems, are well known in patients with sleep apnea. Increasing evidence supports the pathophysiological role of sleep apnea in cardiovascular disorders, which are the leading cause of death in ESRD patients. Uremic factors may be involved in the pathogenesis of sleep apnea in this patient population and optimal dialysis may reduce disease severity. Furthermore, treatment with continuous positive airway pressure may improve quality of life and may help to manage hypertension in these patients. Secondary restless legs syndrome is highly prevalent in patients on maintenance dialysis. The pathophysiology of the disorder may also involve uremia-related factors, iron deficiency, and anemia, but genetic and lifestyle factors might also play a role. The treatment of restless legs syndrome involves various pharmacologic approaches and might be challenging in severe cases. In this article we review the diagnosis and treatment of sleep apnea and restless legs syndrome, with a focus on dialysis patients. We also briefly review current data regarding sleep problems after transplantation, since these studies may indirectly shed light on the possible pathophysiological role of uremia or dialysis in the etiology of sleep disorders. Considering the importance of sleep disorders, more awareness among professionals involved in the care of patients on dialysis is necessary. Appropriate management of sleep disorders could improve the quality of life and possibly even impact upon survival of renal patients. PMID- 16689973 TI - Dialyzer reuse--part II: advantages and disadvantages. AB - Although single dialyzer use and reuse by chemical reprocessing are both associated with some complications, there is no definitive advantage to either in this respect. Some complications occur mainly at the first use of a dialyzer: a new cellophane or cuprophane membrane may activate the complement system, or a noxious agent may be introduced to the dialyzer during production or generated during storage. These agents may not be completely removed during the routine rinsing procedure. The reuse of dialyzers is associated with environmental contamination, allergic reactions, residual chemical infusion (rebound release), inadequate concentration of disinfectants, and pyrogen reactions. Bleach used during reprocessing causes a progressive increase in dialyzer permeability to larger molecules, including albumin. Reprocessing methods without the use of bleach are associated with progressive decreases in membrane permeability, particularly to larger molecules. Most comparative studies have not shown differences in mortality between centers reusing and those not reusing dialyzers, however, the largest cluster of dialysis-related deaths occurred with single-use dialyzers due to the presence of perfluorohydrocarbon introduced during the manufacturing process and not completely removed during preparation of the dialyzers before the dialysis procedure. The cost savings associated with reuse is substantial, especially with more expensive, high-flux synthetic membrane dialyzers. With reuse, some dialysis centers can afford to utilize more efficient dialyzers that are more expensive; consequently they provide a higher dose of dialysis and reduce mortality. Some studies have shown minimally higher morbidity with chemical reuse, depending on the method. Waste disposal is definitely decreased with the reuse of dialyzers, thus environmental impacts are lessened, particularly if reprocessing is done by heat disinfection. It is safe to predict that dialyzer reuse in dialysis centers will continue because it also saves money for the providers. Saving both time for the patient and money for the provider were the main motivations to design a new machine for daily home hemodialysis. The machine, developed in the 1990s, cleans and heat disinfects the dialyzer and lines in situ so they do not need to be changed for a month. In contrast, reuse of dialyzers in home hemodialysis patients treated with other hemodialysis machines is becoming less popular and is almost extinct. PMID- 16689974 TI - Gene polymorphism association studies in dialysis: anemia and host immunity. AB - The typical complications of end-stage renal disease (ESRD) vary in frequency and severity. Whereas anemia of chronic kidney disease might require high doses of exogenous erythropoietin (EPO) in some individuals, others hardly ever need hormone replacement. The same observation can be made for immune defense functions in patients with ESRD. Our understanding of the functional consequences of genetic polymorphisms in a wide array of genes has recently gained momentum. In patients with ESRD, interindividual differences in anemia parameters have been linked in part to genetic polymorphisms. Indeed, although inflammation is an important predictor of the need for EPO, to date a link to cytokine gene polymorphisms has not been established. However, the need for EPO has been linked to polymorphisms in the angiotensin-converting enzyme (ACE) and vitamin D receptor genes. In contrast, in dialysis patients, interindividual variability in host immune responses, including nonspecific (bacterial) and antigen-specific (viral) immune mechanisms has been linked to variations in the interleukin-10 and myeloperoxidase genes as well as variability in human leukocyte antigens (HLAs). In conclusion, the role of genetic polymorphisms in predicting the development and severity of anemia in chronic kidney disease as well as host immune competence is in its infancy and warrants further inquiry. PMID- 16689975 TI - Gene polymorphism association studies in dialysis: bone and mineral metabolism. AB - Abnormalities in bone and mineral metabolism have a significant impact on morbidity and mortality among patients with end-stage renal disease (ESRD). In addition to confounding environmental factors, genetic susceptibility factors may also influence the occurrence and severity of these abnormalities and account for interindividual variability among patients. Indeed, polymorphisms involving genes of the calcium/parathyroid hormone/calcitriol axis have been associated with bone and mineral metabolism abnormalities. This review summarizes studies involving polymorphisms of candidate genes and their effects on the development of complications related to bone and mineral metabolism abnormalities among patients with ESRD. PMID- 16689976 TI - Parathyroid growth and suppression in renal failure. AB - In advanced uremia, parathyroid hormone (PTH) levels should be controlled at a moderately elevated level in order to promote normal bone turnover. As such, a certain degree of parathyroid hyperplasia has to be accepted. Uremia is associated with parathyroid growth. In experimental studies, proliferation of the parathyroid cells is induced by uremia and further promoted by hypocalcemia, phosphorus retention, and vitamin D deficiency. On the other hand, parathyroid cell proliferation might be arrested by treatment with a low-phosphate diet, vitamin D analogs, or calcimimetics. When established, parathyroid hyperplasia is poorly reversible. There exists no convincing evidence of programmed parathyroid cell death or apoptosis in hyperplastic parathyroid tissue or of involution of parathyroid hyperplasia. However, even considerable parathyroid hyperplasia can be controlled when the functional demand for increased PTH levels is removed by normalization of kidney function. Today, secondary hyperparathyroidism can be controlled in patients with long-term uremia in whom considerable parathyroid hyperplasia is to be expected. PTH levels can be suppressed in most uremic patients and this suppression can be maintained by continuous treatment with phosphate binders, vitamin D analogs, or calcimimetics. Thus modern therapy permits controlled development of parathyroid growth. When nonsuppressible secondary hyperparathyroidism is present, nodular hyperplasia with suppressed expression of the calcium-sensing receptor (CaR) and vitamin D receptor (VDR) has been found in most cases. An altered expression of some autocrine/paracrine factors has been demonstrated in the nodules. The altered quality of the parathyroid mass, and not only the increased parathyroid mass per se, might be responsible for uncontrollable hyperparathyroidism in uremia and after kidney transplantation. PMID- 16689977 TI - Complex bypasses and fistulas for difficult hemodialysis access: a prospective, single-center experience. AB - The purpose of this article is to describe several complex vascular access procedures and the outcomes achieved with them in 24 patients (mean age 60 years) undergoing hemodialysis in whom all other accesses had failed and neither peritoneal dialysis nor transplantation was possible. Patients underwent either a necklace bypass (n = 5), axillary loop (n = 1), contralateral internal jugular vein bypass (n = 6), femorofemoral crossover bypass (n = 1), superficial femoral vein transposition (n = 5), axillary artery to popliteal vein bypass (n = 5), or femoral artery to right atrium bypass (n = 1). All grafts implanted were 6 mm, internally reinforced prostheses made of expanded polytetrafluoroethylene (Gore Tex Intering Vascular Graft). Postoperatively patients had bimonthly clinical examinations in which the thrill, bruit, skin, cannulation sites, and adequacy of dialysis were reviewed. A bimonthly ultrasound dilution assessment that included estimation of the graft inflow rate, recirculation rate, and cardiac output was also performed. There was one serious postoperative complication: rapid-onset severe steal syndrome that required immediate tie off of the fistula. During the median follow-up time of 22 months, three patients died of causes unrelated to their vascular access. Nineteen dilatations and 10 surgical revisions were done. Primary patency rates were 83%, 63.5%, and 63.5%, respectively, at 6 months, 1 year, and 2 years; secondary patency rates were 91%, 77%, and 77%. Complex vascular access procedures can provide patients some additional good-quality time on hemodialysis. PMID- 16689978 TI - Response to cyclosporine A in a patient with pure red cell aplasia due to antierythropoietin-alpha antibodies. AB - Reported cases of pure red cell aplasia (PRCA) from the administration of erythropoietin (EPO)-alpha molecule in the United States are rare, and the optimal treatment is still unknown. We present a patient with end-stage renal disease (ESRD) who became hyporesponsive and later unresponsive to EPO-alpha treatment a few months after initiation of hemodialysis. A comprehensive anemia examination was negative while the patient became transfusion dependent. The diagnosis of EPO-alpha-induced PRCA was confirmed by bone marrow biopsy, by undetectable serum EPO levels following the administration of a large dose of EPO alpha, and by documenting the presence of EPO-neutralizing antibodies. Administration of cyclosporine A in addition to prednisone enabled the patient to become transfusion and EPO independent. This case further documents the possible occurrence of PRCA with EPO-alpha administration in the United States and reaffirms the potential beneficial effect of cyclosporine A. PMID- 16689979 TI - Retroperitoneal bleed from acquired renal cysts. PMID- 16689980 TI - Hemoglobin variability: random fluctuation, epiphenomenon, or phenomenon? PMID- 16689981 TI - Alwall's often forgotten contribution to hemodialysis. PMID- 16689984 TI - LRTae: improving statistical power for genetic association with case/control data when phenotype and/or genotype misclassification errors are present. AB - BACKGROUND: In the field of statistical genetics, phenotype and genotype misclassification errors can substantially reduce power to detect association with genetic case/control studies. Misclassification also can bias population frequency parameters such as genotype, haplotype, or multi-locus genotype frequencies. These problems are of particular concern in case/control designs because, short of repeated sampling, there is no way to detect misclassification errors. We developed a double-sampling procedure for case/control genetic association using a likelihood ratio test framework. Different approaches have been proposed to deal with misclassification errors. We have chosen the likelihood framework because of the ease with which misclassification probabilities may be incorporated into in the statistical framework and hypothesis testing. The statistic is called the Likelihood Ratio Test allowing for errors (LRTae) and is freely available via software download. RESULTS: We applied our procedure to 10,000 replicates of simulated case/control data in which we introduced phenotype misclassification errors. The phenotype considered is Ankylosing Spondylitis (AS). The LRTae method power was always greater than LRTstd power for the significance levels considered (5%, 1%, 0.1%, 0.01%). Power gains for the LRTae method over the LRTstd method increased as the significance level became more stringent. Multi-locus genotype frequency estimates using LRTae method were more accurate than estimates using LRTstd method. CONCLUSION: The LRTae method can be applied to single-locus genotypes, multi-locus genotypes, or multi-locus haplotypes in a case/control framework and can be more powerful to detect association in case/control studies when both genotype and/or phenotype errors are present. Furthermore, the LRTae method provides asymptotically unbiased estimates of case and control genotype frequencies, as well as estimates of phenotype and/or genotype misclassification rates. PMID- 16689985 TI - Ethnobotanical investigations among tribes in Madurai District of Tamil Nadu (India). AB - BACKGROUND: An ethnobotanical survey was carried out to collect information on the use of medicinal plants in Southern Western Ghats of India (Madurai district, Tamil Nadu). Information presented in this paper was gathered from the paliyar tribes using an integrated approach of botanical collections, group discussions and interviews with questionnaires in the years 1998 - 1999. The informants interviewed were 12 among whom 4 were tribal practitioners. RESULTS: A total of 60 ethnomedicinal plant species distributed in 32 families are documented in this study. The medicinal plants used by paliyars are listed with Latin name, family, local name, parts used, mode of preparation and medicinal uses. Generally, fresh part of the plant was used for the preparation of medicine. CONCLUSION: We observed that the documented ethnomedicinal plants were mostly used to cure skin diseases, poison bites, stomachache and nervous disorders. The results of this study showed that these tribal people still depend on medicinal plants in Madurai district forest areas. PMID- 16689986 TI - Outcome following patent ductus arteriosus ligation in premature infants: a retrospective cohort analysis. AB - BACKGROUND: The patent ductus arteriosus (PDA) is an important problem in premature infants. Surgical PDA ligation is usually only be considered when medical treatment has either failed or was contraindicated. The aims of our study were to determine the mortality and morbidity following patent ductus arteriosus ligation in premature infants, and whether prostaglandin synthetase inhibitor (PSI) use prior to ligation affects outcome. METHODS: A retrospective case note review study to determine the outcome of premature infants undergoing patent ductus arteriosus ligation in one tertiary neonatal intensive care unit and two paediatric cardiothoracic centres. RESULTS: We had follow-up data on 87 infants. Cumulative mortality rates at 7 days, 30 days and at hospital discharge were 2%, 8% and 20% respectively. The incidence of chronic lung disease, intraventricular haemorrhage, necrotising enterocolitis and retinopathy of prematurity were 77%, 39%, 26% and 28% respectively. There was no difference in mortality, incidence of chronic lung disease or duration of oxygen dependence between those who had and those who had not received a PSI prior to surgical ligation. In those who had received 2 or more courses of PSI prior to surgical ligation, there was a trend to increase in the duration of oxygen therapy and chronic lung disease, but no difference in mortality. CONCLUSION: This study shows that patent ductus arteriosus ligation is a relatively safe procedure (30 day survival 92%) but there is substantial late mortality and a high incidence of morbidity in the survivors. 2 or more courses of PSI prior to surgical ligation trends to increased oxygen dependence and chronic lung disease. This high risk population requires careful follow-up. A definitive prospective cohort study is lacking. PMID- 16689987 TI - A prospective multi-center cohort study of acute non-displaced fractures of the scaphoid: operative versus non-operative treatment [NCT00205985]. AB - BACKGROUND: Acute scaphoid fractures are common in active adults and do lead to reasonable time lost to work. One important goal of treatment is early return to work or sport. On this background, the adequate treatment of non-displaced acute scaphoid fractures is still under discussion. The aim of this study is to compare time to return to previous activity level comparing surgical versus non-surgical treatment of non-displaced acute scaphoid fractures. METHODS/DESIGN: The study is designed as a non-randomized multiple center cohort study including 12 sites in Germany and Austria. The inclusion period is planned to be 12 months with a follow up of 6 months. Allocation to operative or non-operative treatment is chosen by the patient together with his treating surgeon. The primary outcome is time to return to previous activity level adapted for loading of the wrist in daily life as measured by a newly developed questionnaire (PLDL-wrist). Factors identified a priori to be associated with the outcome, e.g., poverty status, age, education, smoking status, gender, and occupation, are measured to ensure adequate control for their potential confounding effects. DISCUSSION: The rationale and the design of a multiple center cohort study are presented. As it is not considered feasible to randomize patients in this study, potential confounding effects need to be controlled adequately. PMID- 16689988 TI - Hospitalisation among immigrants in Italy. AB - BACKGROUND: Immigration is increasing in Italy. In 2003, 2.6 million foreign citizens lived in the country; 52% were men and the majority were young adults who migrated for work. The purpose of this study was to investigate differences in hospitalisation between immigrants and the resident population during the year 2000 in the Lazio region. METHODS: Hospital admissions of immigrants from Less Developed Countries were compared to those of residents. We measured differences in hospitalisation rates and proportions admitted. RESULTS: Adult immigrants have lower hospitalisation rates than residents (134.6 vs. 160.5 per thousand population for acute care; 26.4 vs. 38.3 for day care). However, hospitalisation rates for some specific causes (injuries, particularly for men, infectious diseases, deliveries and induced abortions, ill-defined conditions) were higher for immigrants than for residents. Immigrants under 18 years seem to be generally healthy; causes of admission in this group are similar to those of residents of the same age (respiratory diseases, injuries and poisoning). The only important differences are for infectious and parasitic diseases, with a higher proportion among immigrant youths. CONCLUSION: The low hospitalisation rates for foreigners may suggest that they are a population with good health status. However, critical areas, related to poor living and working conditions and to social vulnerability, have been identified. Under-utilisation of services and low day care rates may be partially due to administrative, linguistic, and cultural barriers. As the presence of foreigners becomes an established phenomenon, it is important to evaluate their epidemiological profile, develop instruments to monitor and fulfil their specific health needs and plan health services for a multi-ethnic population. PMID- 16689989 TI - Issues of informed consent for intrapartum trials: a suggested consent pathway from the experience of the Release trial [ISRCTN13204258]. AB - Service users within the NHS are increasingly being asked to participate in clinical research. In Liverpool Women's NHS Foundation Trust, approximately 35% of women take part in research during their pregnancy. For many studies the consent process is simple; information is provided and signed consent is given. There is a difficulty, however, with obtaining informed consent from women in pregnancy who become eligible only when they develop unforeseen complications, especially when they occur acutely. The problem is compounded with women in labour who may be frightened, vulnerable, in pain, under the effect of opiate analgesia, or all of the above. If research to improve the care of these women is to continue, then special consent procedures are needed. These procedures must ensure that the woman's autonomy is protected whilst recognising that women under these circumstances vary enormously, both in their desire for information and their ability to comprehend it. This paper will discuss the obtaining of consent in this situation, and describe an information and consent pathway for intrapartum research which has been developed in collaboration with consumer groups as a way in which these issues can be tackled. PMID- 16689990 TI - The fading of reported effectiveness. A meta-analysis of randomised controlled trials. AB - BACKGROUND: The "real" effect size of a medical therapy is constant over time. In contrast, the effect size reported in randomised controlled trials (RCTs) may change over time because the sum of all kinds of bias influencing the reported effectiveness is not necessarily constant. As this would affect the validity of meta-analyses, we tested the hypothesis that the reported effect size decreases over time. Furthermore, we tested three hypotheses that would explain a possible change. METHODS: Because of well established outcome measures, the lipid-lowering drugs Pravastatin and Atorvastatin (serum low-density lipoprotein cholesterol, LDL-C) and the anti-glaucoma drugs Timolol and Latanoprost (intraocular pressure, IOP) were chosen for this investigation. Studies were identified by a standardized MEDLINE search. RCTs investigating the above identified medications administered as monotherapy, and in defined dosages, were included. Publication year, baseline (= pre-treatment value in the treatment group of interest) and post intervention means, number of patients and the assignment to experimental or control group were extracted for each study. RESULTS: A total of 625 citations were screened; 206 met the inclusion criteria. The reported effect size of Pravastatin (change of reported effect size in five years: -3.22% LDL-C, P < .0001), Timolol (-0.56 mmHg, P < .0001) and Latanoprost (-1.78 mmHg, P = .0074) decreased over time, while there was no significant change for Atorvastatin (+0.31% LDL-C, P = .8618). Multiple regression analysis showed that baseline values were the most important influencing factor; study size or treatment group did not play a significant role. CONCLUSION: The effectiveness of medical therapies reported in RCTs decreases over time in three of the four investigated pharmaceuticals, caused mainly by baseline differences. We call this phenomenon "fading of reported effectiveness". Under this condition the validity of a meta analysis may be impaired. Therefore we propose to observe this phenomenon in future meta-analyses in order to guarantee a maximum of transparency. PMID- 16689991 TI - Severe reversible cardiac failure after bortezomib treatment combined with chemotherapy in a non-small cell lung cancer patient: a case report. AB - BACKGROUND: Bortezomib (Velcade), a dipeptide boronate proteasome inhibitor, is a novel anti-cancer agent registered for multiple myeloma (MM). It has also shown promising clinical activity in non-small cell lung cancer (NSCLC). Clinical experience with bortezomib so far indicates that overall incidence of cardiac failure associated with bortezomib therapy remains incidental. Nevertheless, acute development or exacerbation of congestive cardiac failure has been associated with bortezomib treatment. CASE PRESENTATION: We present here a case of severe, but reversible, congestive cardiac failure in a lung cancer patient who had no prior cardiac history, after receiving an experimental treatment of bortezomib combined with chemotherapy. Elevated levels of N-terminal pro-B-type natriuretic peptide (NT-proBNP), as retrospectively measured in archived serum samples, were suggestive of pre-existent (sub-clinical) left ventricular dysfunction. CONCLUSION: Based on literature, we hypothesize that baseline presence of sub clinical cardiomyopathy, characterized by a dysregulation of the ubiquitin-proteasome system, could have predisposed this patient for a cardiac side effect induced by systemic proteasome inhibition. Patients with heart disease or risk factors for it should be closely monitored when being submitted to treatment with proteasome inhibition therapy such as bortezomib. Caution is therefore warranted in lung cancer patients who often present with cardiac comorbidities. PMID- 16689992 TI - Web-based climate information resources for malaria control in Africa. AB - Malaria remains a major public health threat to more than 600 million Africans and its control is recognized as critical to achieving the Millennium Development Goals. The greatest burden of malaria in Africa occurs in the endemic regions where the disease pathogen is continuously present in the community. These regions are characterized by an environment that is conducive to interactions between the Anopheles mosquito, malaria parasites and human hosts, as well as housing of generally poor quality, which offers little protection from mosquito human contact. Epidemic malaria tends to occur along the geographical margins of endemic regions, when the equilibrium between the human, parasite and mosquito vector populations is occasionally disturbed and a sharp but temporary increase in disease incidence results. When malaria control measures are inadequate, as is the case in much of sub-Saharan Africa, the disease distribution is closely linked with seasonal patterns of the climate and local environment. In the absence of good epidemiological data on malaria distribution in Africa, climate information has long been used to develop malaria risk maps that illustrate the boundaries of 'climatic suitability for endemic transmission.' The best known of these are produced by the Pan-African-based MARA Collaboration. This paper describes the development of additional malaria suitability maps which have been produced in an online, interactive format to enable temporal information (i.e., seasonality of climate conditions) to be queried and displayed along with spatial information. These maps and the seasonal information that they contain should be useful to the malaria control and health service communities for their planning and operational activities. PMID- 16689993 TI - Reactive oxygen species: role in the development of cancer and various chronic conditions. AB - Oxygen derived species such as superoxide radical, hydrogen peroxide, singlet oxygen and hydroxyl radical are well known to be cytotoxic and have been implicated in the etiology of a wide array of human diseases, including cancer. Various carcinogens may also partly exert their effect by generating reactive oxygen species (ROS) during their metabolism. Oxidative damage to cellular DNA can lead to mutations and may, therefore, play an important role in the initiation and progression of multistage carcinogenesis. The changes in DNA such as base modification, rearrangement of DNA sequence, miscoding of DNA lesion, gene duplication and the activation of oncogenes may be involved in the initiation of various cancers. Elevated levels of ROS and down regulation of ROS scavengers and antioxidant enzymes are associated with various human diseases including various cancers. ROS are also implicated in diabetes and neurodegenerative diseases. ROS influences central cellular processes such as proliferation a, apoptosis, senescence which are implicated in the development of cancer. Understanding the role of ROS as key mediators in signaling cascades may provide various opportunities for pharmacological intervention. PMID- 16689994 TI - Ontogenetic variations in the venom proteome of the Amazonian snake Bothrops atrox. AB - BACKGROUND: Bothrops atrox is responsible for the majority of snakebite accidents in the Brazilian Amazon region. Previous studies have demonstrated that the biological and pharmacological activities of B. atrox venom alter with the age of the animal. Here, we present a comparative proteome analysis of B. atrox venom collected from specimens of three different stages of maturation: juveniles, sub adults and adults. RESULTS: Optimized conditions for two-dimensional gel electrophoresis (2-DE) of pooled venom samples were achieved using immobilized pH gradient (IPG) gels of non-linear 3-10 pH range during the isoelectric focusing step and 10-20% gradient polyacrylamide gels in the second dimension. Software assisted analysis of the 2-DE gels images demonstrated differences in the number and intensity of spots in juvenile, sub-adult and adult venoms. Although peptide mass fingerprinting (PMF) failed to identify even a minor fraction of spots, it allowed us to group spots that displayed similar peptide maps. The spots were subjected to a combination of tandem mass spectrometry and Mascot and MS BLAST database searches that identified several classes of proteins, including metalloproteinases, serine proteinases, lectins, phospholipases A2, L-amino oxidases, nerve growth factors, vascular endothelial growth factors and cysteine rich secretory proteins. CONCLUSION: The analysis of B. atrox samples from specimens of different ages by 2-DE and mass spectrometry suggested that venom proteome alters upon ontogenetic development. We identified stage specific and differentially expressed polypeptides that may be responsible for the activities of the venom in each developmental stage. The results provide insight into the molecular basis of the relation between symptomatology of snakebite accidents in humans and the venom composition. Our findings underscore the importance of the use of venoms from individual specimen at various stages of maturation for the production of antivenoms. PMID- 16689995 TI - Inhibition of NF-kappaB activation by 5-lipoxygenase inhibitors protects brain against injury in a rat model of focal cerebral ischemia. AB - BACKGROUND: Stroke is one of the leading causes of death worldwide and a major cause of morbidity and mortality in the United States of America. Brain ischemia reperfusion (IR) triggers a complex series of biochemical events including inflammation. Leukotrienes derived from 5-lipoxygenase (5-LOX) cause inflammation and are thus involved in the pathobiology of stroke injury. METHODS: To test the neuroprotective efficacy of 5-LOX inhibition in a rat model of focal cerebral IR, ischemic animals were either pre- or post-treated with a potent selective 5-LOX inhibitor, (N- [3-[3-(-fluorophenoxy) phenyl]-1-methyl-2-propenyl]-N-hydroxyurea (BW-B 70C). They were evaluated at 24 h after reperfusion for brain infarction, neurological deficit score, and the expression of 5-LOX. Furthermore, the mechanism and the anti-inflammatory potential of BW-B 70C in the regulation of nuclear factor kappa B (NF-kappaB) and inflammatory inducible nitric oxide synthase (iNOS) were investigated both in vivo and in vitro. RESULTS AND DISCUSSION: Both pre- and post-treatment with BW-B 70C reduced infarctions and improved neurological deficit scores. Immunohistochemical study of brain sections showed IR-mediated increased expression of 5-LOX in the neurons and microglia. BW B 70C down-regulated 5-LOX and inhibited iNOS expression by preventing NF-kappaB activation. Two other structurally different 5-LOX inhibitors were also administered post IR: caffeic acid and 2,3,5-trimethyl-6-[12-hydroxy-5,10 dodecadiynyl]-1,4-benzoquinone (AA-861). As with BW-B 70C, they provided remarkable neuroprotection. Furthermore, in vitro, BW-B 70C inhibited lipopolysaccharide (LPS) mediated nitric oxide production, iNOS induction and NF kappaB activation in the BV2 microglial cell line. Treating rat primary microglia with BW-B70C confirmed blockage of LPS-mediated translocation of the p65 subunit of NF-kappaB from cytosol to nucleus. CONCLUSION: The study demonstrates the neuroprotective potential of 5-LOX inhibition through down-regulation of NF kappaB in a rat model of experimental stroke. PMID- 16689996 TI - Expression of eicosanoid receptors subtypes and eosinophilic inflammation: implication on chronic rhinosinusitis. AB - BACKGROUND: Eicosanoid receptors are G-protein-coupled receptors playing an important immunomodulatory role in airway diseases. However, there is little information on the expression of these receptors and their link with eosinophilic inflammation in paranasal sinus diseases. We aimed with this study to investigate the tissue expression of leukotrienes and prostaglandin E2 receptors in chronic rhinosinusitis patients and the link of this regulation with eosinophilic inflammation. METHODS: Samples were prepared from nasal tissue of patients with chronic rhinosinusitis without nasal polyps (CRS, n = 11), with nasal polyps (CRS NP, n = 13) and healthy subjects (Controls, n = 6). mRNA expression of CysLT1, CysLT2, BLT1, BLT2, E-prostanoid receptors (EP1, EP2, EP3, EP4) and sol-IL 5Ralpha was determined by real-time PCR. Concentrations of PGE2, LTC4/D4/E4, LTB4 and sol-IL-5Ralpha were determined by ELISA and of ECP by ImmunoCap. Protein expression and tissue localization of eicosanoid receptors and activated eosinophils were evaluated by immunohistochemistry. RESULTS: CysLT1 mRNA expression was significantly increased in CRS-NP compared to CRS and controls, and CRS compared to controls, whereas CysLT2 mRNA was enhanced in both CRS groups without differences between them. Levels of both receptors correlated to the number of activated eosinophils, sol-IL-5Ralpha, ECP and LTC4/D4/E4 concentrations in the disease groups. PGE2 protein concentrations and prostanoid receptors EP1 and EP3 were down-regulated in the CRS-NP tissue vs. CRS and controls, whereas EP2 and EP4 expression was enhanced in CRS and CRS-NP patients vs. controls. No differences in BLT receptors were observed between patients and controls. CONCLUSION: CyLTs receptors are up-regulated in nasal polyp tissue and their expression correlate with eosinophilic inflammation supporting previous results. Eicosanoid receptors mRNA pattern observed suggests that down-regulation of EP1 and EP3 in CRS-NP and up-regulation EP2 and EP4 in CRS and CRS-NP groups may have some role in the development of the diseases and their regulation may not be directly linked to eosinophil activation but involve post-transcriptional events mainly related to other inflammatory cell sources. PMID- 16689997 TI - Thermography imaging during static and controlled thermoregulation in complex regional pain syndrome type 1: diagnostic value and involvement of the central sympathetic system. AB - BACKGROUND: Complex Regional Pain Syndrome type 1 (CRPS1) is a clinical diagnosis based on criteria describing symptoms of the disease. The main aim of the present study was to compare the sensitivity and specificity of calculation methods used to assess thermographic images (infrared imaging) obtained during temperature provocation. The secondary objective was to obtain information about the involvement of the sympathetic system in CRPS1. METHODS: We studied 12 patients in whom CRPS1 was diagnosed according to the criteria of Bruehl. High and low whole body cooling and warming induced and reduced sympathetic vasoconstrictor activity. The degree of vasoconstrictor activity in both hands was monitored using a videothermograph. The sensitivity and specificity of the calculation methods used to assess the thermographic images were calculated. RESULTS: The temperature difference between the hands in the CRPS patients increases significantly when the sympathetic system is provoked. At both the maximum and minimum vasoconstriction no significant differences were found in fingertip temperatures between both hands. CONCLUSION: The majority of CRPS1 patients do not show maximal obtainable temperature differences between the involved and contralateral extremity at room temperature (static measurement). During cold and warm temperature challenges this temperature difference increases significantly. As a result a higher sensitivity and specificity could be achieved in the diagnosis of CRPS1. These findings suggest that the sympathetic efferent system is involved in CRPS1. PMID- 16689998 TI - Histone H4 lysine 16 acetylation breaks the genome's silence. AB - Acetylation at histone H4 lysine 16 is involved in many cellular processes in organisms as diverse as yeast and humans. A recent biochemical study pinpoints this particular acetylation mark as a switch for changing chromatin from a repressive to a transcriptionally active state. PMID- 16689999 TI - Relevance of the stroma and epithelial-mesenchymal transition (EMT) for the rheumatic diseases. AB - Epithelial-mesenchymal transition (EMT) is a term applied to the process whereby cells undergo a switch from an epithelial phenotype with tight junctions, lateral, apical, and basal membranes, and lack of mobility into mesenchymal cells that have loose interactions with other cells, are non-polarized, motile and produce an extracellular matrix. The importance of this process was initially recognized from a very early step in embryology, but more recently as a potential mechanism for the progression and spread of epithelial cancers. As the sequence of morphological changes has become understood in molecular terms, diseases characterized by alterations in stromal elements and fibrosis are being considered as examples of EMT. This review will focus on the pathogenetic features of immune-mediated renal disease, systemic sclerosis and rheumatoid arthritis that could be explained by EMT. PMID- 16690000 TI - The seroepidemiology of pertussis in Australia during an epidemic period. AB - Studying the epidemiology of pertussis and impact of differing vaccine schedules is difficult because of differing methods of case ascertainment. The advent of internationally standardized serological diagnosis for recent infection has allowed comparison of age-specific pertussis infection among European countries and was applied in Australia at the time of a major national epidemic. In 1997 and 1998, a nationally representative serum bank using residual sera from diagnostic laboratories was established. Measurement of pertussis toxin (PT) IgG level was conducted by a reference laboratory using an enzyme-linked immunosorbent assay standardized for a number of European countries. A titre of 125 EU/ml was interpreted as indicative of recent pertussis infection. The serological data were correlated with age, gender, region and disease epidemiology in Australia. The highest prevalence of recent pertussis infection was in the 5-9 years age group, and the lowest in 1-4 and 25-64 years age groups. In the 5-14 years age group, 29.7% (5-9 years) and 14.6% (10-14 years) of the sample had serological evidence of recent infection, correlating with the pattern of epidemic notifications. The 15- to 24-year-olds had similar high titres but the same notification rate as 25- to 44-year-olds, suggesting ascertainment bias may result in under-notification in the former age group. The prevalence of high titres observed was up to 20-fold higher than some European countries during a similar time period. Although vaccination has reduced the transmission of pertussis in the youngest and most vulnerable age group, pertussis is still endemic in Australia, particularly in older children and the elderly. The Australian vaccination schedule has been changed in an attempt to address this problem, by spacing doses more widely, with the fifth dose at 15-17 years of age. Seroepidemiology for pertussis offers the potential to compare patterns of pertussis between countries and examine the impact of vaccine schedule changes independent of notification and diagnostic bias. PMID- 16690001 TI - The epidemiology of leptospirosis and the emergence of Leptospira borgpetersenii serovar Arborea in Queensland, Australia, 1998-2004. AB - Leptospirosis is one of the most commonly encountered zoonoses in both Australia and the rest of the world. The incidence of leptospirosis in Queensland over the 7-year study period (1998-2004) was 3.1/100000 population. Enhanced surveillance questionnaires were used to collect patient data and facilitate an epidemiological investigation of leptospirosis in Queensland. Farming occupations comprised the majority of occupational exposure cases, however, recreational exposure accounted for 18% of the 883 cases. Rainfall and the presence of animal hosts had the most influence on the incidence of leptospirosis. Several trends in serovar numbers over this period are noted, in particular the emergence of L. borgpetersenii serovar Arborea, which accounted for 22% of all leptospirosis cases in Australia and 68% of South-East Queensland cases in 2004. Assessment of epidemiological trends in leptospirosis is important to obtain directed public health intervention and outcomes in the reduction of leptospirosis cases. PMID- 16690002 TI - Outbreaks of waterborne infectious intestinal disease in England and Wales, 1992 2003. AB - We reviewed the epidemiological and microbiological characteristics of 89 reported outbreaks of waterborne infectious intestinal disease affecting 4321 people in England and Wales over the period 1992-2003. Public water supplies were implicated in 24 outbreaks (27%), private water supplies in 25 (28%), swimming pools in 35 (39%) and other sources in five outbreaks (6%). Cryptosporidium was implicated in 69% of outbreaks, Campylobacter sp. in 14%, Giardia in 2%, E. coli O157 in 3% and Astrovirus in 1%. From 2000, there was a consistent decline in the number of outbreaks of waterborne disease associated with public water supplies. The incidence rate of outbreaks in recipients of private water supplies may be as high as 35 times the rate in those receiving public water supplies (1830 vs. 53 per million population). Private water suppliers need to be aware of the importance of adequate treatment and the prevention of faecal contamination of storage water. Swimming-pool operators need to ensure chlorination and in particular adequate filtration measures are in place. PMID- 16690003 TI - Number of deaths among HIV-infected adults in France in 2000, three-source capture-recapture estimation. AB - We estimated the number of deaths in France for the year 2000 in HIV-infected adults using three sources. The sources were (1) the 'Mortalite 2000' survey (M2000): 964 deaths were documented by 185 hospital wards involved in HIV management; (2) 1288 death certificates with a mention of HIV infection (INSERM CepiDc) and (3) the French hospital database on HIV infection (FHDH) identified 654 deaths. The capture-recapture method was used with log-linear modelling. Overall 1559 deaths were observed. Estimation of the number of deaths in France was 1699 (95% CI 1671-1727). The completeness of M2000, CepiDc and FHDH were 55%, 76% and 38% respectively. Diversification of diseases and of causes of death in HIV-infected adults may explain: (1) the diversification of physicians involved in their management and incomplete coverage of M2000 and FHDH, and (2) why HIV infection was not mentioned in all death certificates. PMID- 16690004 TI - Epidemiology and genetic characterization of Shigella flexneri strains isolated from three paediatric populations in Egypt (2000-2004). AB - Ninety-seven isolates of Shigella flexneri from children seeking medical care from three sites in Egypt were characterized. Overall, 46.4% of children (median age 17 months) were febrile or reported blood in their stools, 25.8% were dehydrated and 16.5% were admitted to hospital. Serotypes 2a (37.1%), 1b (18.6%), 1c (17.5%), and 6 (15.5%) comprised over 88.7% of the total isolates. We observed marked resistance to ampicillin (87.6%), tetracycline (84.5%) and trimethoprim sulfamethoxazole (63.9%). Pulsed-field electrophoresis grouped the majority of isolates within a serotype together, separately from isolates of an alternative serotype. The set gene was present in all serogroup 2a isolates, however, the sen gene was detected in every isolate. Our results show S. flexneri 1c has emerged as a dominant S. flexneri serotype in Egypt. Development and application of a Shigella vaccine should consider the diversity of Shigella serotypes within a geographical region prior to administration. PMID- 16690005 TI - Effects of different doses of venlafaxine on serotonin and norepinephrine reuptake in healthy volunteers. AB - Venlafaxine is generally considered to be a dual 5-HT and NE reuptake inhibitor when it is used at doses above 75 mg/d in humans. While its 5-HT reuptake inhibiting property has been demonstrated, some controversy still exists regarding the doses of venlafaxine required to inhibit NE reuptake. Healthy male volunteers received, on a double-blind basis, paroxetine (20 mg/d), desipramine (100 mg/d), nefazodone (300 mg/d), or venlafaxine (150 or 300 mg/d) in the last 5 d of a 7-d period of administration. Inhibition of 5-HT reuptake was estimated by determining the degree of depletion of whole-blood 5-HT, while that of NE was assessed by measuring the attenuation of the systolic blood pressure increases produced by intravenous injections of tyramine. Paroxetine, both regimens of venlafaxine, and to a lesser extent desipramine significantly decreased whole blood 5-HT content. Nefazodone failed to produce any significant change. Desipramine abolished the tyramine pressor response, whereas all other drug regimens left this parameter unaltered. Venlafaxine and paroxetine acted as potent 5-HT reuptake inhibitors in the present study. In contrast, neither the moderate nor the high dose of venlafaxine displayed any significant inhibiting activity in this model assessing NE reuptake in peripheral NE terminals. The validity of the model was confirmed by the potent inhibitory action of desipramine on NE reuptake. While the reasons for this unexpected lack of action remain unclear, venlafaxine appeared to be an effective NE reuptake agent in depressed patients using the same approach. PMID- 16690006 TI - Differential physiological effects of a low dose and high doses of venlafaxine in major depression. AB - Venlafaxine is an antidepressant drug with demonstrated serotonin (5-HT) and norepinephrine (NE) reuptake blockade properties in electrophysiological and microdialysis experiments in laboratory animals. In healthy volunteers, its 5-HT reuptake-inhibiting potential has also been clearly documented, but not its NE reuptake blockade action. This double-blind study compared the effects of a low dose (75 mg) and of a forced titration of high (up to 375 mg in 1 wk) daily doses of venlafaxine. Forty-four patients with major depression according to DSM-IV criteria were assessed bi-weekly for the first 2 wk and weekly for the next 2 wk. Inhibition of 5-HT reuptake was estimated using the depletion of whole-blood 5 HT, while that of NE was assessed using the attenuation of the systolic blood pressure elevations produced by intravenous injections of tyramine. Forty-two patients completed the study. Both the low and the high doses of venlafaxine decreased the levels of 5-HT to the same extent: the reduction was of about 55% after 1 wk and of 75% after 4 wk. The 75 mg/d dose of venlafaxine did not alter the tyramine pressor response, whereas, in patients receiving the higher regimens of venlafaxine, there was a significant attenuation of the pressor effect of tyramine. There was no significant difference between the two treatment arms regarding the modifications of the depression scores. The present data showed that, at its minimal effective dose in depression (75 mg/d), venlafaxine acted as a selective 5-HT reuptake inhibitor, whereas when administered at higher doses (225 and 375 mg/d), it acted as a dual 5-HT and NE reuptake inhibitor. PMID- 16690007 TI - Pathophysiology and therapy for haemoglobinopathies. Part I: sickle cell disease. AB - In sickle cell disease, a single base pair substitution in the gene encoding the beta-globin chain of the haemoglobin molecule gives rise to a surprisingly broad spectrum of pathophysiological and clinical manifestations. Inflammation, endothelial activation, red blood cell membrane abnormalities and altered availability of vasoactive factors characterise this disorder. Clinically, patients suffer from a host of seemingly unrelated maladies, from pain episodes to strokes, life-threatening infections and pulmonary hypertension. Deepened understanding of this complex disease now allows us to begin to turn away from simple supportive treatments, and move towards therapies aimed at specific pathophysiological targets. This article, the first of two reviews on the pathophysiology of haemoglobinopathies, discusses the molecular basis of sickle cell disease, and elaborates on the many factors that exacerbate or ameliorate the disease process. It then focuses on the promising targeted therapies currently in use or under investigation. An accompanying article on haemoglobinopathies (Part II) focuses on thalassaemias. PMID- 16690008 TI - Silencing the snorers: no gain without pain? AB - OBJECTIVE: To compare the estimated effects of conservative surgery to those of a mandibular advancement splint (MAS) in the treatment of habitual snoring. METHOD: Prospective, observational, non-randomized cohort study. RESULTS: Adequate follow up data were available for 88 participants (23 following coblation, 65 after provision of an MAS). The mean reduction in snoring symptoms inventory (SSI) score for the 23 subjects undergoing coblation was 9.83 (+/-standard deviation 11.43). Reported pain duration was greatest with uvula amputation, but uvula coblation did not always adequately reduce its bulk. Of the 65 MAS patients, 39 (60 per cent) used the device regularly, with a mean fall in SSI of 12 (+/-16.4, p=0.001). Approximately one in four patients in both groups achieved a significant fall in SSI (>15 points), and the measured effect sizes were close to 0.75 for both treatments. CONCLUSIONS: Mandibular advancement splints and coblation have similar efficacies. However, their efficacy does not match that of radical surgery. PMID- 16690010 TI - MDP and other muropeptides--direct and synergistic effects on the immune system. AB - Muropeptides are breakdown products of peptidoglycan (PGN) of Gram-negative and Gram-positive bacteria. They are released during bacterial growth and division, as part of the host response by lysozyme and amidases, or upon antibiotic treatment. After phagocytosis of bacteria or bacterial breakdown products by host immune cells, the muropeptides trigger intracellular signaling cascades, leading to altered gene expression and activation of the immune response. Numerous muropeptides and derivatives have been synthesized chemically to characterize their immunostimulatory effects and adjuvant activity. Muramyl dipeptide, a natural partial structure of PGN, is the minimal structure with adjuvant activity. This review discusses the structure and occurrence of muropeptides and gives a broad overview of their inflammatory and adjuvant activity and the possible involvement of receptors in these responses. PMID- 16690011 TI - Heat stress response results in increased macrophage inflammatory protein-2 concentration in a lipopolysaccharide-exposed macrophage cell line. AB - Pretreatment with heat confers cardiopulmonary protection in endotoxemic animals. This mechanism may be through suppression of pro-inflammatory mediator production. The objectives of this study were to determine the effect of heat stress on tumor necrosis factor-alpha (TNF-alpha) and macrophage inflammatory protein-2 (MIP-2) in a lipopolysaccharide-exposed macrophage cell line and to study the relationship between TNF-alpha and MIP-2 production. Heat pretreatment resulted in decreased TNF-alpha transcription and translation by lipopolysaccharide-exposed macrophages; and increased MIP-2 concentration without additional effect in transcription. Administration of TNF-alpha antibody prior to exposure to lipopolysaccharide resulted in increased MIP-2 concentration suggesting that TNF-alpha acts to down-regulate MIP-2 production. The mechanism by which heat stress causes an increase in MIP-2 concentration may be secondary to its suppressing effect on TNF-alpha production. PMID- 16690012 TI - Stable isotope metabolic labeling of Neisseria meningitidis lipooligosaccharide. AB - The lipooligosaccharide (LOS) of a Neisseria meningitidis acetate auxotroph was metabolically labeled with either [2-13C]-sodium acetate or [1,2-13C2]-sodium acetate. In this study, we demonstrated that this label was efficiently incorporated into both the lipid A acyl moieties and the two N-acetylglucosamines present in the oligosaccharide branch of the LOS. The development of this efficient labeling protocol should prove useful in future structural studies analyzing the interactions between LOS and host proteins. PMID- 16690013 TI - Inflammatory, coagulatory and circulatory responses to logarithmic increases in the endotoxin dose in the anaesthetised pig. AB - Although porcine intravenous endotoxin shock models are widely employed in experimental sepsis, endotoxin dose-effect studies are scarce. Our primary aim was to establish the dose response to increasing endotoxin doses in inflammatory, coagulatory and haemodynamic effect variables, as well as to determine the optimal time point for assessment in a pig model. A secondary aim was to study pathophysiological covariations between the different responses. Twenty anaesthetised piglets received endotoxin intravenously in doses of 0.063 (n = 3), 0.25 (n = 3), 1.0 (n = 3), 4.0 (n = 3), 8 (n = 3) and 16 microg/kg/h (n = 2). In addition, non-endotoxin piglets constituted a control group (n = 3). Physiological variables were registered and blood samples analysed for TNF-alpha, IL-6, leukocyte, platelet and haemoglobin concentrations hourly for 6 h. Increases in the endotoxin dose induced significant log-log cytokine responses as well as log-linear leukocyte and platelet responses. Significant log-linear responses were observed for circulatory parameters, plasma leakage, hypoperfusion and pulmonary compliance. Significant covariations in the responses were noted. In conclusion, there were log-log or log-linear responses to endotoxin suggesting a greater effect of a given dose at lower pre-existing endotoxin concentrations and lower doses of < or = 1 microg/kg/h may be of advantage in experiments designed to study potential anti-endotoxin effects of experimental drugs or measures. PMID- 16690014 TI - Safety of incremental inhaled lipopolysaccharide challenge in humans. AB - BACKGROUND: Inhalation of environmental endotoxin is important in the pathogenesis of asthma and other environmental airway diseases. Inhaled airway challenge using lipopolysaccharide in humans has been performed for over 20 years to assess the airway response to endotoxin. However, there are no published data on the short-term safety of endotoxin inhalation protocols. OBJECTIVE: To characterize the safety and tolerability of incremental inhaled lipopolysaccharide challenge in humans. PATIENTS AND METHODS: We performed a retrospective analysis of data obtained from 119 subjects who underwent inhaled challenge with up to 41.5 mug of lipopolysaccharide. We measured pulmonary function, temperature, mean arterial pressure, heart rate, and systemic symptoms for 3 h after challenge. RESULTS: Fever occurred in 30% of subjects and was associated with a higher cumulative dose of lipopolysaccharide. Reduced mean arterial pressure occurred in 21% of subjects and was dose-related. There was no association between fever or decreased mean arterial pressure and airway responsiveness to inhaled lipopolysaccharide. Common symptoms reported by subjects included: chills (64%), malaise (56%), cough (56%), chest tightness (49%), headache (43%), and myalgias (27%). None of the subjects experienced delayed discharge or a serious adverse event. CONCLUSIONS: Inhaled lipopolysaccharide causes dose-related systemic responses that include fever, reduced blood pressure, and constitutional symptoms that are not associated with the airway response to inhaled lipopolysaccharide. Systemic responses to inhaled lipopolysaccharide should be expected and subjects undergoing inhaled lipopolysaccharide challenge in the research setting should be carefully monitored for non-pulmonary adverse events for several hours after challenge. PMID- 16690015 TI - Localization of the mouse defense lectin ficolin B in lysosomes of activated macrophages. AB - Ficolins are pattern-recognition molecules of the innate immune system able to trigger the lectin pathway of the complement activation upon binding to microbial surfaces. In humans, two plasma ficolins have been identified and characterized, whereas a third cell-associated ficolin (M-ficolin) was found on monocyte surfaces. The mouse homologue of M-ficolin is called ficolin B. Although the spatial-temporal expression patterns of mouse ficolins have been described recently, the subcellular localization of ficolin B protein is so far unknown. By using ficolin B-specific antibodies and confocal microscopy, we show that ficolin B is expressed within mouse peritoneal exudate macrophages and is co-localized with Lamp-1, a marker for lysosomes and late endosomes. In addition, the data indicate that ficolin B expression is up-regulated upon macrophage activation. PMID- 16690017 TI - [Von Meyenburg disease found during laparoscopic surgery: report of two cases]. AB - The authors report two cases of von Meyenburg complexes disease found during laparoscopic surgical procedures. The first patient (41 years old) had multiple gallbladder stones with recurrent hepatic colics. The second patient (45 years old) had gastroesophageal reflux with recurrent peptic esophagitis. In both cases, multiple millimetric and superficial hepatic lesions were found during the exploration of the abdominal cavity. Then, an hepatic biopsy was done because the macroscopic aspect should suspect secondary metastatic lesions. Histological result made the diagnosis of von Meyenburg complexes disease (biliary microhamartomas). It seems to be important to know this anomaly of the ductal plate development and to do a liver biopsy during laparoscopy to affirm the diagnosis and cancel the diagnosis of multiple hepatic metastasis. In fact, this congenital pathology could be associated with increased risk of cholangiocarcinoma of the liver. The modality of radiological monitoring still remains to be defined. PMID- 16690018 TI - [Sentinel node mapping in anorectal melanoma]. AB - Anorectal melanoma is a rare condition and its surgical management is controversial. This article reports the case of a patient with anorectal melanoma who underwent abdominoperineal resection and Sentinel Lymph Node biopsy. Melanoma was classified pT4aN0. Fifty months after initial treatment, the patient is still alive disease free. SLN mapping allows better surgical excision of the presumed sites of the lymphatic dissemination in melanoma. SLN biopsy improve the accuracy of nodal staging. In case of sentinel node metastasis, it allows early therapeutic lymphadenectomy of the sentinel nodes's basin and could therefore reduce the high rate of regional recurrence in anorectal melanoma. Moreover, knowing the exact histological status of the regional nodes means that the relative merits of abdominoperineal resection and wild local excision could be compared in relation to tumor thickness. PMID- 16690019 TI - Promotion of hair follicle development and trichogenesis by Wnt-10b in cultured embryonic skin and in reconstituted skin. AB - We previously showed that Wnt-10b promoted the differentiation of primary skin epithelial cells (MPSEC) toward hair shaft and inner root sheath of the hair follicle (IRS) cells in vitro. In the present study, we found that Wnt-10b promotes the development of hair follicles using a culture of mouse embryonic skin tissue and trichogenesis using a reconstitution experiment with nude mice. Hair follicle development was observed in skin taken from mouse embryos on embryonic day 10.5 following a 2-day culture with recombinant Wnt-10b (rWnt-10b), however, not without rWnt-10b. Brown hair growth was observed at the site of reconstituted skin in Balb/c nude mice where dermal fibroblasts and keratinocytes, derived from C3H/HeN new born mice, were transplanted with Wnt-10b producing COS cells (Wnt-COS). Without the co-transplantation of Wnt-COS, no hair growth was observed. Our results suggest an important role of Wnt-10b in the initiation of hair follicle development and following trichogenesis. PMID- 16690020 TI - Enhancement of adipose tissue formation by implantation of adipogenic differentiated preadipocytes. AB - Engineered adipose tissue could be used for the reconstruction or augmentation of soft tissues lost due to mastectomy or lumpectomy in plastic and reconstructive surgery. Preadipocytes are a feasible cell source for adipose tissue regeneration. However, the enhancement of the in vivo adipogenic conversion of preadipocytes remains a major task. In vitro, the adipogenic differentiation of preadipocytes prior to implantation might enhance the adipose tissue regeneration. In the present study, we investigated whether implantation of adipogenic-differentiated preadipocytes enhances the adipose tissue formation compared with implantation of undifferentiated preadipocytes. We also investigated whether basic fibroblast growth factor (bFGF) further enhances the adipose tissue formation mediated by the implantation of adipogenic differentiated preadipocytes. A fibrin matrix containing human preadipocytes cultured in adipogenic differentiation-inducing conditions with (group 1) or without (group 2) bFGF was injected into the subcutaneous spaces of athymic mice. Fibrin matrices containing undifferentiated human preadipocytes with (group 3) or without (group 4) bFGF were also implanted. Six weeks after implantation, the implanted cells formed new tissues in all groups. Importantly, the implantation of adipogenic-differentiated preadipocytes resulted in more extensive adipogenesis than the implantation of undifferentiated preadipocytes, as evaluated by adipose tissue area and human adipocyte-specific gene expression in the newly formed tissues. In addition, bFGF enhanced neovascularization in the newly formed tissues and further enhanced the adipogenesis mediated by the adipogenic-differentiated preadipocytes. The present study demonstrates that the implantation of adipogenic-differentiated preadipocytes enhances adipose tissue regeneration, as compared with the implantation of undifferentiated preadipocytes, and that cell transplantation-mediated adipogenesis can be further enhanced by the delivery of bFGF. PMID- 16690021 TI - Isolation and properties of the CYP2D25 promoter: transcriptional regulation by vitamin D3 metabolites. AB - Previous studies have suggested that hepatic production of 25-hydroxyvitamin D3 may be suppressed by 1alpha,25-dihydroxyvitamin D3. However, the molecular details of these observations have not been clarified. In the current study, the 5'-flanking DNA sequence of CYP2D25, a porcine microsomal vitamin D 25 hydroxylase, was isolated and analyzed. The CYP2D25 promoter contains a putative vitamin D response element (VDRE). The promoter activity was markedly suppressed by 1alpha,25-dihydroxyvitamin D3 and 25-hydroxyvitamin D3 in presence of vitamin D receptor (VDR). The data suggest that VDR-mediated inhibition of 25 hydroxylase(s) by vitamin D3 metabolites at the transcriptional level may play an important role in the regulation of 25-hydroxyvitamin D3 production in liver and other tissues. PMID- 16690022 TI - Validation of housekeeping genes as internal control for studying gene expression in rice by quantitative real-time PCR. AB - For accurate and reliable gene expression results, normalization of real-time PCR data is required against a control gene, which displays highly uniform expression in living organisms during various phases of development and under different environmental conditions. We assessed the gene expression of 10 frequently used housekeeping genes, including 18S rRNA, 25S rRNA, UBC, UBQ5, UBQ10, ACT11, GAPDH, eEF-1alpha, eIF-4a, and beta-TUB, in a diverse set of 25 rice samples. Their expression varied considerably in different tissue samples analyzed. The expression of UBQ5 and eEF-1alpha was most stable across all the tissue samples examined. However, 18S and 25S rRNA exhibited most stable expression in plants grown under various environmental conditions. Also, a set of two genes was found to be better as control for normalization of the data. The expression of these genes (with more uniform expression) can be used for normalization of real-time PCR results for gene expression studies in a wide variety of samples in rice. PMID- 16690023 TI - TcRho1, the Trypanosoma cruzi Rho homologue, regulates cell-adhesion properties: evidence for a conserved function. AB - Rho proteins are members of the Ras superfamily of small GTPases. In higher eukaryotes these proteins play pivotal role in cell movement, phagocytosis, intracellular transport, cell-adhesion, and maintenance of cell morphology, mainly through the regulation of actin microfilaments. The GTPase TcRho1 is the only member of the Rho family described in human protozoan parasite Trypanosoma cruzi. We previously demonstrated that TcRho1 is actually required for differentiation of epimastigote to trypomastigote forms during the parasite cell cycle. In the present work, we describe cellular phenotypes induced by TcRho1 heterologous expression in NIH 3T3 fibroblasts. The NIH-3T3 lineages expressing the TcRho1-G15V and TcRho1-Q76L mutants displayed decreased levels of migration compared to the control lineage NIH-3T3 pcDNA3.1, a phenotype probably due to distinct cell-substrate adhesion properties expressed by the mutant cell lines. Accordingly, cell-substrate adhesion assays revealed that the mutant cell lines of NIH-3T3 expressing TcRho1-positive dominants constructions present enhanced substrate-adhesion phenotype. Furthermore, similar experiments with T. cruzi expressing TcRho1 mutants also revealed an enhancement of cell attachment. These results suggest that TcRho1 plays a conserved regulatory role in cell-substrate adhesion in both NIH-3T3 fibroblasts and T. cruzi epimastigotes. Taken together, our data corroborate the notion that TcRho1 may regulate the substrate-adhesion in T. cruzi, a critical step for successful progression of the parasite life cycle. PMID- 16690024 TI - NAADP+ synthesis from cADPRP and nicotinic acid by ADP-ribosyl cyclases. AB - ADP-ribosyl cyclases (ADPRCs) are present from lower Metazoa to mammals and synthesize the Ca2+-active (di)nucleotides cyclic ADP-ribose (cADPR), NAADP+, and ADP-ribose (ADPR), involved in the regulation of important cellular functions. NAADP+ can be synthesized by ADPRCs from NADP+ through a base-exchange reaction, which substitutes nicotinamide for nicotinic acid (NA). Here we demonstrate that ADPRCs from both lower and higher Metazoa (including human CD38) can also synthesize NAADP+ starting from 2'-phospho-cyclic ADP-ribose (cADPRP) and NA. Comparison, on the two substrates cADPRP and NADP+, of the relative rates of the reactions introducing NA and hydrolyzing/cyclizing the substrate, respectively, indicates that with all ADPRCs tested cADPRP is preferentially transformed into NAADP+, while NADP+ is preferentially cyclized or hydrolyzed to cADPRP/2'-phospho ADP-ribose. cADPRP was detectable in retinoic acid-differentiated, CD38+ HL-60 cells, but not in undifferentiated, CD38- cells. These results suggest that cADPRP may be a NAADP+ precursor in ADPRC+ cells. PMID- 16690025 TI - Intestinal and peri-tumoral lymphatic endothelial cells are resistant to radiation-induced apoptosis. AB - Radiation therapy is a widely used cancer treatment, but it is unable to completely block cancer metastasis. The lymphatic vasculature serves as the primary route for metastatic spread, but little is known about how lymphatic endothelial cells respond to radiation. Here, we show that lymphatic endothelial cells in the small intestine and peri-tumor areas are highly resistant to radiation injury, while blood vessel endothelial cells in the small intestine are relatively sensitive. Our results suggest the need for alternative therapeutic modalities that can block lymphatic endothelial cell survival, and thus disrupt the integrity of lymphatic vessels in peri-tumor areas. PMID- 16690026 TI - Expression of a human anti-rabies virus monoclonal antibody in tobacco cell culture. AB - A Nicotiana tabacum cv. Xanthi cell culture was initiated from a transgenic plant expressing a human anti-rabies virus monoclonal antibody. Within 3 months, plant cell suspension cultures were established and recombinant protein expression was examined. The antibody was stably produced during culture growth. ELISA, protein G purification, Western blotting, and neutralization assay confirmed that the antibody was fully processed, with association of light and heavy-chains, and that it was able to bind and neutralize rabies virus. Quantification of antibody production in plant cell suspension culture revealed 30 microg/g of cell dry weight for the highest-producing culture (0.5 mg/L), 3 times higher than from the original transgenic plant. The same production level was observed 3 months after cell culture initiation. Plant cell suspension cultures were successfully grown in a new disposable plastic bioreactor, with a growth rate and production level similar to that of cultures in Erlenmeyer flasks. PMID- 16690027 TI - Mechanism of fibroblast growth factor-binding protein 1 repression by TGF-beta. AB - Transforming growth factor-beta (TGF-beta) is the prototypical member of a family of growth factors that play important roles in normal development and human diseases. We identified the gene for fibroblast growth factor-binding protein 1 (FGF-BP1) as being significantly repressed following TGF-beta treatment. FGF-BP1 is an extracellular matrix bound protein that enhances fibroblast growth factor (FGF) signaling. We demonstrate here that TGF-beta signaling significantly represses FGF-BP1 expression in mesenchymal and neural crest cells undergoing in vitro smooth muscle differentiation. Analysis of the downstream signaling pathways shows that Smad2/3 are crucial for efficient FGF-BP1 repression by TGF beta. Furthermore, we identified a novel element in the region from -785 to -782 bp of the FGF-BP1 promoter, which represents a known binding site for Hypermethylation in Cancer-1 (Hic-1), necessary for repression of FGF-BP1 by TGF beta. These data define the molecular mechanism of transcriptional repression of an important target of TGF-beta signaling during angiogenesis. PMID- 16690028 TI - Caspase-dependant activation of chymotrypsin-like proteases mediates nuclear events during Jurkat T cell apoptosis. AB - Apoptosis involves a cascade of biochemical and morphological changes resulting in the systematic disintegration of the cell. Caspases are central mediators of this process. Supporting and primary roles for serine proteases as pro-apoptotic mediators have also been highlighted. Evidence for such roles comes largely from the use of pharmacological inhibitors; as a consequence information regarding their apoptotic function and biochemical properties has been limited. Here, we circumvented limitations associated with traditional serine protease inhibitors through use of a fluorescently labelled inhibitor of serine proteases (FLISP) that allowed for analysis of the specificity, regulation and positioning of apoptotic serine proteases within a classical apoptotic cascade. We demonstrate that staurosporine triggers a caspase-dependant induction of chymotrypsin-like activity in the nucleus of apoptotic Jurkat T cells. We show that serine protease activity is required for the generation of late stage nuclear events including condensation, fragmentation and DNA degradation. Furthermore, we reveal caspase dependant activation of two chymotrypsin-like protein species that we hypothesize mediate cell death-associated nuclear events. PMID- 16690029 TI - Possible role of LRP1, a CCN2 receptor, in chondrocytes. AB - Low density lipoprotein receptor (LDLR)-related protein 1 (LRP1/CD91) is one of the receptors of CCN2 that conducts endochondral ossification and cartilage repair. LRP1 is a well-known endocytic receptor, but its distribution among chondrocytes remains to be elucidated. We herein demonstrate for the first time that the distribution of LRP1 in chondrocytes except for hypertrophic chondrocytes in vivo and in vitro. Interestingly, the LRP1 levels were higher in mature chondrocytic HCS-2/8 and osteoblastic SaOS-2 than in other cells, whereas the other LDLR family members involved in ossification were detected at lower levels in HCS-2/8. It was interesting to note that in HCS-2/8, LRP1 was observed not only on the cell surface and in the cytoplasm, but also in the nucleus. Exogenously added CCN2 was incorporated into HCS-2/8, which was partially co localized with LRP1, and targeted to the recycling endosomes and nucleus as well as the lysosomes. These findings suggest specific roles of LRP1 in cartilage biology. PMID- 16690030 TI - Action of apoptotic endonuclease DNase gamma on naked DNA and chromatin substrates. AB - The internucleosomal cleavage of genomic DNA is a biochemical hallmark of apoptosis. DNase gamma, a Mg2+/Ca2+-dependent endonuclease, has been suggested to be one of the apoptotic endonucleases, but its biochemical characteristic has not been fully elucidated. Here, using recombinant DNase gamma, we showed that DNase gamma is a Mg2+/Ca2+-dependent single-stranded DNA nickase and has a high activity at low ionic strength. Under higher ionic strength, such as physiological buffer conditions, the endonuclease activity of DNase gamma is restricted, but its activity is enhanced in the presence of linker histone H1, which explains DNA cleavage at linker regions of apoptotic nuclei. PMID- 16690031 TI - Alternative 3' UTR polyadenylation of Bzw1 transcripts display differential translation efficiency and tissue-specific expression. AB - BZW1 is a conserved regulatory factor for transcriptional control of histone H4 gene at the G1/S transition. In this study, three Bzw1 transcripts were identified in mice with two long forms (approximately 2.9 kb) expressed ubiquitously at low level, and a short transcript of 1.8 kb expressed at high level exclusively in testis. These different transcripts share the same 5' UTR and coding sequence, but differ in the length of 3' UTR by utilizing alternative polyadenylation sites. Different translation efficiencies were observed in the cells transfected with chimeric EGFP-Bzw1 genes tailed with different 3' UTRs. Our results demonstrate that Bzw1 transcripts are alternatively polyadenylated and expressed in tissue-specific pattern. PMID- 16690032 TI - Characterization of histone (H1B) oxalate binding protein in experimental urolithiasis and bioinformatics approach to study its oxalate interaction. AB - The rat kidney H1 oxalate binding protein was isolated and purified. Oxalate binds exclusively with H1B fraction of H1 histone. Oxalate binding activity is inhibited by lysine group modifiers such as 4',4'-diisothiostilbene-2,2 disulfonic acid (DIDS) and pyridoxal phosphate and reduced in presence of ATP and ADP. RNA has no effect on oxalate binding activity of H1B whereas DNA inhibits oxalate binding activity. Equilibrium dialysis method showed that H1B oxalate binding protein has two binding sites for oxalate, one with high affinity, other with low affinity. Histone H1B was modeled in silico using Modeller8v1 software tool since experimental structure is not available. In silico interaction studies predict that histone H1B-oxalate interaction take place through lysine121, lysine139, and leucine68. H1B oxalate binding protein is found to be a promoter of calcium oxalate crystal (CaOx) growth. A 10% increase in the promoting activity is observed in hyperoxaluric rat kidney H1B. Interaction of H1B oxalate binding protein with CaOx crystals favors the formation of intertwined calcium oxalate dehydrate (COD) crystals as studied by light microscopy. Intertwined COD crystals and aggregates of COD crystals were more pronounced in the presence of hyperoxalauric H1B. PMID- 16690033 TI - Integrated analysis of multiple data sources reveals modular structure of biological networks. AB - It has been a challenging task to integrate high-throughput data into investigations of the systematic and dynamic organization of biological networks. Here, we presented a simple hierarchical clustering algorithm that goes a long way to achieve this aim. Our method effectively reveals the modular structure of the yeast protein-protein interaction network and distinguishes protein complexes from functional modules by integrating high-throughput protein-protein interaction data with the added subcellular localization and expression profile data. Furthermore, we take advantage of the detected modules to provide a reliably functional context for the uncharacterized components within modules. On the other hand, the integration of various protein-protein association information makes our method robust to false-positives, especially for derived protein complexes. More importantly, this simple method can be extended naturally to other types of data fusion and provides a framework for the study of more comprehensive properties of the biological network and other forms of complex networks. PMID- 16690034 TI - SNPs on chips: the hidden genetic code in expression arrays. AB - Gene expression microarray analysis in postmortem brains is one of the fastest growing fields of psychiatric research. Here we show that common polymorphisms (SNPs) present on probe sets can masquerade as significant "gene expression" differences. After first observing this artifact in the Catechol-O-methyl transferase (COMT) gene, we replicate the finding in two additional genes predicted to show this artifact. Many Affymetrix chips contain thousands of SNPs that are both common and in the central probe region affecting hybridization, and thus have the potential to confound expression analysis. PMID- 16690035 TI - Developmental expression of Bis protein in the cerebral cortex and hippocampus of rats. AB - Bis (Bcl-2 interacting death suppressor), identified as a Bcl-2-binding protein, has been suggested to have diverse functions in addition to binding to Bcl-2, thereby regulating cell death. To investigate the potential role of Bis in the developing brain, the spatiotemporal expression of Bis protein was studied in the rat forebrain during prenatal and early postnatal development using immunohistochemistry. Initial expression of Bis was detected in the medial telencephalic wall of the lateral ventricle, the area most likely corresponded to the cortical hem from the earliest age examined (E13). There was an abrupt increase of immunoreactive neurons in the cortex and hippocampus during the first postnatal week, which declined thereafter. Two populations of Bis-immunoreactive neurons can be clearly distinguished in the developing forebrain: a population of differentiating and postmitotic neurons coexpressing Bis and microtubule associated protein-2 (MAP-2), and a population of neurons with the characteristic morphology of Cajal-Retzius cells located exclusively in the marginal zone/layer I of the cortex and in the hippocampal equivalents of the marginal zone. The latter neurons were colabeled with reelin, a marker for Cajal-Retzius cells. While Bis expression in the cerebral cortex and hippocampus exists only transiently by P14, considerable expression was found to be maintained in the rostral migratory stream and the subventricular zone of the lateral ventricle, where Bis-immunoreactive cells were glutamine synthetase-positive glial cells. Our results suggest that Bis may contribute to the developmental processes, including the differentiation and maturation of specific neuronal populations in relation to Bcl-2 in the developing rat forebrain. PMID- 16690036 TI - Laminin activates CaMK-II to stabilize nascent embryonic axons. AB - In neurons, the interaction of laminin with its receptor, beta1 integrin, is accompanied by an increase in cytosolic Ca2+. Neuronal behavior is influenced by CaMK-II, the type II Ca2+/calmodulin-dependent protein kinase, which is enriched in axons of mouse embryonic neurons. In this study, we sought to determine whether CaMK-II is activated by laminin, and if so, how CaMK-II influences axonal growth and stability. Axons grew up to 200 microm within 1 day of plating P19 embryoid bodies on laminin-1 (EHS laminin). Activated CaMK-II was found enriched along the axon and in the growth cone as detected using a phospho-Thr(287) specific CaMK-II antibody. beta1 integrin was found in a similar pattern along the axon and in the growth cone. Direct inhibition of CaMK-II in 1-day-old neurons immediately froze growth cone dynamics, disorganized F-actin and ultimately led to axon retraction. Collapsed axonal remnants exhibited diminished phospho-CaMK-II levels. Treatment of 1-day neurons with a beta1 integrin-blocking antibody (CD29) also reduced axon length and phospho-CaMK-II levels and, like CaMK-II inhibitors, decreased CaMK-II activation. Among several CaMK-II variants detected in these cultures, the 52-kDa delta variant preferentially associated with actin and beta 3 tubulin as determined by reciprocal immunoprecipitation. Our findings indicate that persistent activation of delta CaMK-II by laminin stabilizes nascent embryonic axons through its influence on the actin cytoskeleton. PMID- 16690037 TI - Gestational nicotine-induced changes in adolescent neuronal activity. AB - Smoking during pregnancy is associated with numerous physiological and neurobehavioral deficits in infants, which persist into adolescence. To better understand the underlying mechanisms, we have treated pregnant rats with nicotine and have evaluated expression of the immediate early gene c-fos, as a measure of neuronal activity, in the brains of adolescent male offspring. Pregnant dams were infused with nicotine (3 mg/kg/day) or saline from gestational day (G) 4 until G18. After birth on G22, litters were cross fostered and weaned at postnatal day (P) 21. Brain sections from adolescent offspring, aged P38-40, were analyzed by in situ hybridization for regional c-fos mRNA expression in response to acute injection of saline or nicotine (0.03, 0.1, 0.3 mg/kg). Acute nicotine challenge increased c-fos expression within nucleus accumbens shell, lateral bed nucleus of the stria terminalis, paraventricular nucleus of the hypothalamus, dorsal lateral geniculate, and superior colliculus, whereas c-fos expression was decreased in prelimbic cortex. There was no effect of gestational nicotine treatment on acute nicotine-induced alterations in c-fos mRNA levels. However, basal c-fos mRNA expression within infralimbic cortex and nucleus accumbens core was increased by gestational nicotine treatment. These data indicate that gestational nicotine does not produce global changes in nicotine-induced c-fos expression in adolescent brain. However, gestational drug exposure changes basal neuronal activity within mesocorticolimbic structures that are critical for motivated behavior. Such changes may underlie some of the behavioral deficits in attention, cognition, and impulse control that have been reported in the offspring of smoking mothers. PMID- 16690038 TI - Actomyosin-dependent microtubule rearrangement in lysophosphatidic acid-induced neurite remodeling of young cortical neurons. AB - It has been shown that lysophosphatidic acid (LPA), a signaling phospholipid, induces neurite retraction and the formation of retraction fibers in young cortical neurons by actin rearrangement. This study examined the rearrangement of microtubules (MTs) during LPA-induced neurite remodeling by immunostaining with antibodies against several types of tubulin. The results showed that alpha tubulin was present in growing neurites as well as in cell bodies with various localization profiles. Exposure of neurons to LPA resulted in neurite retraction, accompanied by the rearrangement of MTs in neurites and the accumulation of MTs in cell bodies, without significant changes in the total amount of MTs in the cytoskeletal fraction of cultured neurons. Similar findings were obtained when young neurons were stained for other types of tubulin, including beta-tubulin type III and posttranslationally acetylated and tyrosinated tubulin. LPA-induced MT rearrangement was accompanied by accumulation of myosin IIB and polymerized actin at the base of retraction fibers. These effects of LPA on MTs and myosin IIB were blocked by pretreatment with inhibitors of the actomyosin and Rho pathways (cytochalasin D, blebbistatin, and Y27632), but not by an MT stabilizer (taxol), whereas taxol inhibited neurite retraction and MT depolymerization induced by nocodazole. Furthermore, neurofilaments also showed rearrangement in response to LPA, which was blocked by cytochalasin D and Y27632, but not taxol. Taken together, these results suggested that LPA did not induce MT depolymerization and that LPA-induced actomyosin activation produced MT and neurofilament rearrangement, leading to neurite remodeling. PMID- 16690039 TI - Electrocortical correlates of control of selective attention to spatial frequency. AB - In the present study, we investigated control of selective attention to spatial frequency patterns, using a cueing paradigm. Subjects either used the instruction embedded in a word cue to prepare for the upcoming test stimulus (transient attention condition) or used the instruction they received before a block of trials (sustained reference condition), under completely similar stimulus conditions. The pattern of differential cue responses between these two conditions, reflecting top-down attentional control processes, was different between two groups of subjects, effectively canceling each other out. Despite comparable behavioral performance on both cues and targets, one group (n = 4) elicited a fronto-central-parietal positivity, starting 500 ms postcue over frontal and prefrontal areas, later including more central and posterior scalp sites, whereas another group (n = 8) started 400 ms postcue over central sites with a negativity, growing in strength over time and stabilizing over fronto central sites. Only the group of eight subjects showed some evidence of occipital pretarget biasing activity. Independent of group, source modeling of the attentional control activity showed that attentional control was initiated in anterior, not posterior, parts of the brain. Furthermore, different underlying sources were found for both groups, in addition to signs of differential processing of target stimuli. Possible individual differences in attentional control ability and its relation to usage of different brain areas to deal with the task demands are discussed in more detail. PMID- 16690040 TI - Isolation and characterization of putative O2 chemoreceptor cells from the gills of channel catfish (Ictalurus punctatus). AB - Little is known about the cells or mechanisms of O2 chemoreception in vertebrates other than mammals. The purpose of this study, therefore, was to identify O2 sensitive chemoreceptors in a fish. Putative O2-sensitive chemoreceptors were dissociated from the gills of channel catfish, Ictalurus punctatus, and cultured. A population of cells was identified with morphology and a histochemical profile similar to mammalian carotid body Type I (glomus) cells and pulmonary neuroepithelial cells. These cells stain with neutral red and appear to be the branchial neuroepithelial cells. Immunocytochemical staining showed that these cells contain neuron-specific enolase (NSE), tyrosine hydroxylase (TH) and 5 hydroxytryptamine (5HT). Patch-clamp experiments showed that these cells have a O2-sensitive, voltage-dependent outward K+ current like mammalian O2 sensors. Two kinds of electrophysiological responses to hypoxia (P(O2) < 10 Torr) were observed. Some cells showed inhibition of outward current in response to hypoxia, whereas other cells showed potentiation. Neurochemical content and electrophysiological responses to hypoxia indicate that these cells are piscine O2-sensitive chemoreceptors. PMID- 16690041 TI - Structural characteristics and physicochemical properties of oxidized corn starches varying in amylose content. AB - The effects of amylose content on the extent of oxidation and the distribution of carboxyl groups in hypochlorite-oxidized corn starches were investigated. Corn starches including waxy corn starch (WC), common corn starch (CC), and 50% and 70% high-amylose corn starches (AMC) were oxidized with NaOCl at three concentrations (0.8%, 2%, and 5%). Carboxyl and carbonyl content of oxidized starches increased with increasing NaOCl concentration. High-AMC (70%) had slightly higher carboxyl and carbonyl contents at 0.8% NaOCl, whereas WC had significantly higher carboxyl and carbonyl contents at 2% and 5% NaOCl levels. Carbohydrate profiles by high-performance size-exclusion chromatography indicate that amylose was more susceptible to depolymerization than amylopectin. Degradation of amylopectin long chains (DP >24) was more pronounced in WC and CC than in AMCs. The crystalline lamellae of WC started to degrade at 2% NaOCl, but those of the other corn starches remained intact even at 5% NaOCl level according to X-ray crystallinity. By using anion-exchange chromatography for separation and size-exclusion chromatography for characterization, carboxyl groups were found to be more concentrated on amylopectin than on amylose, particularly in AMCs. Oxidation decreased gelatinization temperature and enthalpy with WC showing the most decrease and 70% AMC showing the least. The gelatinization enthalpy of 50% AMC decreased significantly faster than those of CC and 70% AMC after 0.8% oxidation. Retrogradation of amylopectin slightly increased after oxidation with increasing oxidation level. The peak viscosities of oxidized WC and CC were higher than those of their native counterparts at 0.8% NaOCl, but this increase was not observed in AMCs. The setback viscosities of 2% NaOCl-oxidized 50% and 70% AMCs were much higher than those of the unmodified counterparts. The extent of oxidation and physicochemical properties of oxidized starches varied greatly with the amylase:amylopectin ratio of corn starches. Amylose was suggested to play an important role in controlling the oxidation efficiency. PMID- 16690042 TI - JunD attenuates phenylephrine-mediated cardiomyocyte hypertrophy by negatively regulating AP-1 transcriptional activity. AB - OBJECTIVE: Mice deficient for the AP-1 transcription factor JunD, the only Jun protein constitutively expressed and clearly detectable in the mammalian heart, develop enhanced cardiac hypertrophy in response to chronic pressure overload. Catecholamines inducing alpha-adrenergic receptor-mediated signaling have been implicated in the neurohumoral response to pressure overload and the development of left ventricular hypertrophy. In the present study we analyzed the mechanistic role of JunD in cardiomyocyte hypertrophy in vitro in response to alpha adrenergic agonist phenylephrine (PE). METHODS: Cardiomyocytes were isolated from 1- to 3-day-old rats and transfected with adenoviruses expressing LacZ or wild type JunD, or with expression vectors encoding LacZ, wild-type JunD, mutated JunD forming only JunD homodimers (JunDeb1), mutated JunD lacking the JNK site (JunD Delta 162), or c-Jun. After stimulation with PE (10(-5) mol/L), hypertrophic growth of cardiomyocytes (cross-sectional area and [3H]-leucine incorporation) and mRNA expression of JunD, c-Jun, c-Fos, and atrial natriuretic peptide (ANP) were analyzed. Transcriptional activation was determined by luciferase activity in cardiomyocytes transfected with AP-1 or ANP luciferase reporter plasmids. Gel shift assays with an AP-1 consensus oligonucleotide were performed to analyze AP 1 DNA binding activities. RESULTS: PE augmented mRNA levels of c-Jun and c-Fos, but decreased JunD transcript levels. Adenoviral over-expression of wild-type JunD blunted PE-induced hypertrophic growth and expression of ANP mRNA. Over expression of JunD in cardiomyocytes caused enhanced AP-1 protein-DNA binding, without increasing the transcriptional response from AP-1 or ANP luciferase reporter plasmids at baseline or upon PE stimulation. Moreover, over-expression of JunDeb1 attenuated transcription from AP-1 or ANP luciferase reporter plasmids and blunted c-Jun-mediated acceleration of AP-1 transcriptional activity at baseline and in response to PE. CONCLUSIONS: Our observations establish a novel role for JunD as a negative regulator of cardiomyocyte hypertrophy in response to hypertrophic stimuli by inhibiting AP-1 transcriptional activity. PMID- 16690043 TI - Unzipping RyR2 in adult cardiomyocytes: getting closer to mechanisms of inherited ventricular arrhythmias? PMID- 16690044 TI - A new short synthesis of 10R-tuberculostearic acid and its enantiomer. AB - Tuberculostearic acid (10R-methyloctadecanoic acid) and its 10S-enantiomer were synthesised by a chiral pool strategy, in four steps from citronellyl bromide. PMID- 16690046 TI - Excretion of urinary enzymes in normal pregnancy. AB - OBJECTIVES: To study the pattern of excretion of enzymes in urine during normal pregnancy. DESIGN AND METHODS: Primigravidae, with uncomplicated pregnancies, were followed up throughout gestation. Urine samples were collected from them and activities of alanine aminopeptidase (AAP), gamma-glutamyl transferase (GGT), acid phosphatase (ACP) and N-acetyl-beta-d-glucosaminidase (NAG) activities in urine were estimated. RESULTS: Small but significant increases were found in the activities of AAP and NAG excreted through the course of pregnancy. The changes seen in excretion of ACP and GGT were not statistically significant. CONCLUSIONS: Changes in excretion of ACP and GGT may be useful indicators of renal dysfunction in pregnancy, as their activities did not vary significantly through the course of normal pregnancy. PMID- 16690047 TI - The characteristics of elastic fiber assembled with recombinant tropoelastin isoform. AB - OBJECTIVE: It is known that elastin mRNA is transcribed from a single gene. The variety of tropoelastin isoforms results from multiple alternative splicing of the primary transcript. The purpose of this study was to investigate the characteristics of elastic fiber assembled with tropoelastin isoform, which is full-length human tropoelastin (HTE), exon 26A missing tropoelastin (Delta26A), and exon 32 missing tropoelastin (Delta32). DESIGN AND METHODS: We demonstrated the process of elastic fiber assembly and the existence of elastic fiber resistant to pancreatic elastase with HTE, Delta26A, or Delta32 fiber using an in vitro model of elastic fiber assembly. These elastic fibers were evaluated by immunofluorescent staining, the quantitative analysis of cross-linked amino acids, and semi-quantitative analysis of matrix-associated tropoelastin. RESULTS: There were no big differences getting into the matrix among these tropoelastins in immunofluorescence microscopy and semi-quantitative analysis. In the comparison with the HTE, the Delta26A and the Delta32 significantly increased and decreased, respectively, the formation of cross-linking amino acids and the binding to scaffold proteins. Furthermore, it was found that it is difficult to degrade the Delta26A assembly with pancreatic elastase as compared with HTE or Delta32 assembly. CONCLUSION: The elastic fiber assembled with the tropoelastin isoforms was characterized using an in vitro model. The present study provides important information regarding the pathology of human diseases including emphysema and atherosclerosis. PMID- 16690048 TI - Math5 is required for both early retinal neuron differentiation and cell cycle progression. AB - CNS progenitors choose a fate, exit mitosis and differentiate. Basic helix-loop helix (bHLH) transcription factors are key regulators of neurogenesis, but their molecular mechanisms remain unclear. In the mouse retina, removal of the bHLH factor Math5 (Atoh7) causes the loss of retinal ganglion cells (RGCs) and appearance of excess cone photoreceptors. Here, we show a simultaneous requirement for Math5 in retinal neuron formation and cell cycle progression. At embryonic day E11.5, Math5-/- cells are unable to assume the earliest fates, particularly that of an RGC, and instead adopt the last fate as Muller glia. Concurrently, the loss of Math5 causes mitotically active retinal progenitors to undergo aberrant cell cycles. The drastic fate shift of Math5-/- cells correlates with age-specific alterations in p27/Kip1 expression and an inability to become fully postmitotic. Finally, Math5 normally suppresses NeuroD1 within Math5 expressing cells and inhibits Ngn2 expression and cone photoreceptor genesis within separate cell populations. Thus, Math5 orchestrates neurogenesis in multiple ways, regulating both intrinsic and extrinsic processes. PMID- 16690049 TI - Xenopus ADAMTS1 negatively modulates FGF signaling independent of its metalloprotease activity. AB - We have isolated the Xenopus ortholog of ADAMTS1 (a disintegrin and metalloprotease with thrombospondin motifs), XADAMTS1, which is expressed in the presumptive ectoderm, then the Spemann organizer, and later in the trunk organizer region and posterior ectoderm in the Xenopus embryo. We show that, when overexpressed in the dorsal marginal zone or in the anterior ectoderm by mRNA injection, XADAMTS1 inhibits gastrulation or generates embryos with an enlarged cement gland, respectively. XADAMTS1 also reduces the expression of Xbra in both whole embryos and FGF-treated animal caps. These effects of XADAMTS1 are likely to be due to its inhibition of the Ras-MAPK cascade because XADAMTS1 inhibits the phosphorylation of ERK by FGF4 in animal caps. Deletion analysis of XADAMTS1 revealed that a combination of the signal peptide and the C-terminal region containing the thrombospondin type 1 repeats is necessary and sufficient for this function, whereas the metalloprotease domain is dispensable. In addition, loss-of function analysis with antisense morpholino oligos showed that knockdown of XADAMTS1 sensitizes animal caps to Xbra induction by FGF2. These data suggest that secreted XADAMTS1 negatively modulates FGF signaling in the Xenopus embryo. PMID- 16690050 TI - The Toll immune-regulated Drosophila protein Fondue is involved in hemolymph clotting and puparium formation. AB - Clotting is critical in limiting hemolymph loss and initiating wound healing in insects as in vertebrates. It is also an important immune defense, quickly forming a secondary barrier to infection, immobilizing bacteria and thereby promoting their killing. However, hemolymph clotting is one of the least understood immune responses in insects. Here, we characterize fondue (fon; CG15825), an immune-responsive gene of Drosophila melanogaster that encodes an abundant hemolymph protein containing multiple repeat blocks. After knockdown of fon by RNAi, bead aggregation activity of larval hemolymph is strongly reduced, and wound closure is affected. fon is thus the second Drosophila gene after hemolectin (hml), for which a knockdown causes a clotting phenotype. In contrast to hml-RNAi larvae, clot fibers are still observed in samples from fon-RNAi larvae. However, clot fibers from fon-RNAi larvae are more ductile and longer than in wt hemolymph samples, indicating that Fondue might be involved in cross linking of fiber proteins. In addition, fon-RNAi larvae exhibit melanotic tumors and constitutive expression of the antifungal peptide gene Drosomycin (Drs), while fon-RNAi pupae display an aberrant pupal phenotype. Altogether, our studies indicate that Fondue is a major hemolymph protein required for efficient clotting in Drosophila. PMID- 16690051 TI - El Nino Southern Oscillation (ENSO) and dysentery in Shandong province, China. AB - To investigate the impact of the El Nino Southern Oscillation (ENSO) on dysentery transmission, the relationship between monthly dysentery cases in Shandong Province of China and the monthly Southern Oscillation Index (SOI), a broad index of ENSO, was examined over the period 1991-2003. Spearman correlations and generalized linear models were calculated to detect the association between the SOI and dysentery cases. Data from 1991 to 2001 were used to estimate the parameters, while data from 2002 to 2003 were used to test the forecasting ability of the model. After controlling for seasonality, autocorrelation, and a time-lagged effect, the results indicate that there was a significant negative association between the number of dysentery cases and the SOI, with a lagged effect of 2 months. A one-standard-deviation decrease in the SOI might cause up to 207 more dysentery cases per month in Shandong Province. This is the first report of the impact of the Southern Oscillation on dysentery risk in China, indicating that the SOI may be a useful early indicator of potential dysentery risk in Shandong Province. PMID- 16690052 TI - Pharmacological separation between peripheral and central functions of cyclooxygenase-2 with CIAA, a novel cyclooxygenase-2 inhibitor. AB - There are many reports concerning the physiological and pathological involvement of cyclooxygenase (COX)-2 in the central nervous system and peripheral tissue cells. Selective COX-2 inhibitors that mainly distribute peripherally have not been reported thus far. Therefore central and peripheral roles of COX-2 remain classified pharmacologically. In this study, in vivo pharmacological profiles of CIAA ([6-chloro-2-(4-chlorobenzoyl)-1H-indol-3-yl]acetic acid), a novel selective COX-2 inhibitor which distributes at higher concentrations in plasma than in brain, were compared with those of well-known selective COX-2 inhibitors, celecoxib and rofecoxib. Additionally, the possibility of pharmacological separation between peripheral and central actions of COX-2 with the inhibitors was investigated. CIAA selectively inhibited COX-2 activity compared with COX-1 in in vitro assays with rat whole blood. The compound exhibited lower brain penetration and higher plasma concentration (the brain/plasma concentration ratio was approximately 0.02) than celecoxib and rofecoxib after oral administration. Therefore, CIAA is mainly expected to act peripherally. Edema and prostaglandin E2 (PGE2) production in Carrageenan-injected rat paws, and pyrexia and PGE2 production in the brain in lipopolysaccharide (LPS)-injected rats were measured in in vivo experiments. CIAA exhibited lower ratios of anti-pyretic/anti edematous activities and of inhibitory activities of PGE2 production in brain/paw than those of celecoxib and rofecoxib, and these ratios well-reflected brain/plasma concentration ratios. In conclusion, we discovered a novel selective COX-2 inhibitor, CIAA, which distributes at higher concentrations in plasma than in brain, which would make possible the pharmacological separation of the peripheral and central functions of COX-2. PMID- 16690053 TI - Mast cell chymase induces expression of chemokines for neutrophils in eosinophilic EoL-1 cells and mouse peritonitis eosinophils. AB - Human chymase induced release of interleukin-8 (IL-8) in human EoL-1 cells that had been differentiated into eosinophil-like cells with butyric acid. The chymase induced IL-8 production was specific in that other cytokines/chemokines examined were not induced. Human chymase also increased mRNA for IL-8 in the differentiated EoL-1 cells, showing involvement of mRNA synthesis. The chymase induced IL-8 release was inhibited by pertussis toxin as well as U0126 (an inhibitor for extracellular signal-regulated kinase pathway) and SB203580 (p38 inhibitor), suggesting that the chymase-induced IL-8 production is mediated by G protein-coupled receptor and mitogen-activated protein kinases. Mouse mast cell protease-4 (mMCP-4), a mouse chymase, induced macrophage-inflammatory protein-2 (MIP-2), a mouse homologue for IL-8, in mouse eosinophils in vitro. Intradermal injection of mMCP-4 not only induced skin edema but increased MIP-2 content and neutrophil number at the injection site. Taken together, our findings demonstrate that mast cell chymase may contribute to the interaction between eosinophils and neutrophils by inducing IL-8/MIP-2 in eosinophils at allergic inflamed sites. PMID- 16690054 TI - Pactimibe stabilizes atherosclerotic plaque through macrophage acyl CoA:cholesterol acyltransferase inhibition in WHHL rabbits. AB - Novel acyl coenzyme A:cholesterol acyltransferase (ACAT) inhibitor pactimibe was administered as the sulfate salt form to 3-month-old homozygous Watanabe heritable hyperlipidemic (WHHL) rabbits at doses of 0, 10, or 30 mg/kg for 32 weeks. Pactimibe (10 and 30 mg/kg) tended to reduce intimal thickening in thoracic aortic lesions (294+/-39 and 276+/-32 microm, respectively, versus 313+/ 37 microm control), histopathological examination revealing significantly increased smooth muscle cell area (12.0+/-0.9% and 12.3+/-0.5%, P<0.05, respectively, versus 9.7+/-0.8% control), significantly increased collagen fiber area (20.5+/-1.2% and 31.0+/-1.3%, P<0.05, respectively, versus 16.2+/-1.0% control), and tended to reduce macrophage infiltration (6.0+/-1.1% and 4.6+/ 1.0%, respectively, versus 7.0+/-1.3% control). Pactimibe dose-dependently reduced cholesteryl ester content in thoracic and abdominal aortic lesions, and reduced free cholesterol content in the aorta versus control. Although pactimibe did not alter serum cholesterol levels in WHHL rabbits, it stabilized vulnerable plaque characterized with reduced cholesteryl ester content, enriched collagen fibers and increased smooth muscle cells, indicating potential as a treatment strategy for coronary heart disease. PMID- 16690055 TI - Glucose utilization by the retinal pigment epithelium: evidence for rapid uptake and storage in glycogen, followed by glycogen utilization. AB - Glucose utilization and glycogen metabolism by human retinal pigment epithelium (RPE) cultures with high transepithelial resistance maintained on porous Millicell polycarbonate filters, were quantified by fluorophore-assisted carbohydrate electrophoresis (FACE). Glucose uptake was more efficient at the apical surface of the RPE. The utilization of glucose when restricted to either the apical or basal medium was also evaluated. Under both conditions, glucose was quickly transported to the opposite compartment and rapidly utilized. However, glucose from the apical compartment was depleted to a greater extent than from the basal compartment. The de novo synthesis and accumulation of glycogen accompanied glucose utilization. This was paralleled by a concomitant increase in lysosomal glycogen degradation measured as an increase in cell-associated maltodextrins. The highest levels of glucose in glycogen and maltodextrins occurred at 24 h, declining to basal levels at 72 h. Glucose transporter expression in the RPE cultures was evaluated with the reverse transcriptase polymerase chain reaction. Glucose transporter-1 (GLUT 1) was the isoform expressed in these cells. GLUT 1 localization was determined by immunocytochemistry. GLUT 1 localizes to the apical and basolateral border of the RPE. The intensity of fluorescence was higher on the apical border. The rapid depletion of medium glucose suggests that RPE culture studies should replenish medium glucose more frequently than every 72 h to maintain physiologically relevant glucose concentrations. These studies are the first to demonstrate glucose, glycogen and maltodextrin metabolism by RPE cells, and their detection and quantitation by FACE. PMID- 16690056 TI - Visual cortical plasticity following unilateral sensorimotor cortical lesions in the neonatal rat. AB - Previous work has shown that unilateral sensorimotor cortex (SMC) lesions in newborn rats resulted in an apparent shift of the motor cortex map in the spared hemisphere, particularly of the hindlimb cortex. In view of such findings, the present study was initiated to determine if the visual cortex located both ipsilateral and contralateral to neonatal SMC, or contralateral to occipital cortical (OC) lesions, would show similar remodeling. Visual evoked potentials (VEPs) were used to map the visual cortex electrophysiologically. The results show an expansion of the visual cortex, in both the contralateral and ipsilateral hemisphere, into normally motor cortical areas in adult animals that had sustained unilateral neonatal unilateral SMC lesions. In contrast, similar changes were not seen within the spared visual cortex after unilateral occipital cortical lesions, suggesting that the shift in the visual map was specifically in response to the SMC lesion and was not a generalized response to neonatal cortical damage. Histological analysis showed a functional expansion in the rostral boundary of visual cortex with no corresponding cytoarchitectural alterations. PMID- 16690057 TI - Neuroprotection of adenoviral-vector-mediated GDNF expression against kainic-acid induced excitotoxicity in the rat hippocampus. AB - Glial-cell-line-derived neurotrophic factor (GDNF) is a potent survival factor for several types of neurons. In the present study, we examined the protective roles of adenoviral-vector-delivered GDNF (Ad-GDNF) in the hippocampus damaged by kainic-acid (KA)-induced excitotoxicity using GAD-67 immunoreactivity, immunoblot analysis, behavioral test, 5-bromo-2-deoxyuridine (BrdU) and TUNEL assay. Ad-GDNF was pre-inoculated into the KA-treated rat hippocampus 7 days before KA injection. Ad-GDNF resulted in the suppression of KA-induced tonic-clonic convulsions. In situ apoptosis assay demonstrated a significant reduction in apoptotic cells in the CA3 and dentate hilus regions of the Ad-GDNF-pre inoculated rats (Ad-GDNF-KA), compared to the KA rats. Striking reductions in the density of GAD-67 neurons were also observed in the CA3 and dentate hilus regions of the KA rats. On the other hand, the number of GAD-67-positive cells was recovered to the control levels in the Ad-GDNF-KA rats. Immunoblot analysis further confirmed that GAD-67 and Bcl-2 expression increased in the Ad-GDNF-KA rats compared to KA rats. Taken together, these results suggest that Ad-GDNF may serve to control KA-induced hippocampal cell loss and behavioral seizure. PMID- 16690058 TI - Outcome of in vitro fertilization/intracytoplasmic sperm injection after laparoscopic cystectomy for endometriomas. AB - OBJECTIVE: To assess the impact of prior unilateral or bilateral endometrioma cystectomy on controlled ovarian hyperstimulation (COH) and intracytoplasmic sperm injection (ICSI) outcome. DESIGN: Retrospective case-control study. SETTING: Department of Obstetrics and Gynecology, School of Medicine, Hacettepe University, Ankara, Turkey. PATIENT(S): Fifty-seven consecutive infertile patients were enrolled who had previously undergone unilateral (n = 34) or bilateral (n = 23) laparoscopic cystectomy for endometriomas more than 3 cm in diameter and underwent ICSI. The control group consisted of 99 patients with tubal factor infertility. INTERVENTION(S): Controlled ovarian hyperstimulation and ICSI. MAIN OUTCOME MEASURE(S): Cycle cancellation rate, number of oocytes, fertilization rate, embryo quality, clinical pregnancy rate (PR), and implantation rate. RESULT(S): The mean number of oocytes, metaphase II oocytes, and two-pronucleated oocytes were significantly lower in the bilateral cystectomy group compared to the unilateral cystectomy and control groups. However, all other parameters, including fertilization rate, the mean number of embryos transferred, the mean number of grade 1 embryos transferred, the clinical PR per embryo transfer, and implantation rate, were comparable among the three groups. Within the unilateral cystectomy group, the mean number of oocyte retrieved from the operated site was significantly less than in the contralateral nonoperated site. CONCLUSION(S): Laparoscopic endometrioma cystectomy does reduce the ovarian reserve. However, diminished ovarian reserve does not translate into impaired pregnancy outcome. PMID- 16690059 TI - Gonadotropin stimulation demonstrates a ceiling effect on in vitro fertilization outcomes. AB - OBJECTIVE: To determine the effect of number of oocytes retrieved and number of 2PN embryos developed on in vitro fertilization (IVF) outcomes. DESIGN: Retrospective data analysis. SETTING: University practice. PATIENT(S): Reproductive-aged women (n = 467). INTERVENTION(S): First fresh nondonor cycle of IVF. MAIN OUTCOME MEASURE(S): Clinical pregnancy rate RESULT(S): Clinical pregnancy rates increase until age 30 (odds ratio (OR) 1.72 per year (95% confidence interval 1.19-2.49)) before demonstrating a linear decline. In subjects < 37 years old, maximal clinical pregnancy rates are seen when 20 oocytes were retrieved (OR 1.03 (0.96-1.11)), five 2-pronuclei (2PN) embryos developed (OR 1.91 (1.29-2.87)), and no more than two embryos transferred (OR 0.72 (0.56-0.92) for each additional embryo transferred > 2). In subjects > or = 37 years old, maximum clinical pregnancy rates were achieved in subjects who had ten oocytes retrieved (OR 1.09 (1.01-1.18)), 20 2PN embryos developed (OR 1.29 (1.03-1.62)), and no more than two embryos transferred (OR 0.72 (0.56-0.92) for each additional embryo transferred > 2). CONCLUSION(S): The odds of achieving a successful clinical pregnancy with IVF are greatest with retrieval of approximately 20 oocytes, transfer of no more than 2 embryos, and the development of about five 2PN embryos in women < 37 years old and ten 2PN embryos in women > or = 37 years old. PMID- 16690060 TI - Professional self-regulation for preimplantation genetic diagnosis: experience of the American Society for Reproductive Medicine and other professional societies. AB - Professional societies that are stakeholders in preimplantation genetic diagnosis have democratically elected officers and favor self-regulation. Their practice guidelines are cognizant of societal needs and allow for introduction of new technologies. These societies have demonstrated the ability for professional self regulation by collection, validation, and promulgation of center-specific results. PMID- 16690061 TI - Intellectual impairment: a forgotten speciality. PMID- 16690062 TI - Load transmission in the nasofrontal suture of the pig, Sus scrofa. AB - The nasofrontal suture links the nasal complex with the braincase and is subject to compressive strain during mastication and (theoretically) tensile strain during growth of nasal soft tissues. The suture's ability to transmit compressive and tensile loads therefore affects both cranioskeletal stress distribution and growth. This study investigated the in vitro viscoelastic and failure properties of the nasofrontal suture in the pig, Sus scrofa. Suture specimens from two ages were tested in compression and tension and at fast and slow rates. In additional specimens, strain gauges were applied to the suture and nasal bone for strain measurement during testing. Relaxation testing demonstrated higher elastic moduli in tension than compression, regardless of test rate or pig age. In contrast, maximum elastic moduli from failure tests, as well as peak stresses, were significantly higher in compression than in tension. Sutures from older pigs tended to have higher elastic moduli and peak stresses, significantly so for tensile relaxation moduli. Strain gauge results showed that deformation at the suture was much greater than that of the nasal bone. These data demonstrate the viscoelasticity and deformability of the nasofrontal sutural ligament. The suture achieved maximal resistance to tensile deformation at low loads, corresponding with the low tensile loads likely to occur during growth of nasal soft tissues. In contrast, the maximal stiffness in compression at high loads indicates that the suture functions with a substantial safety factor during mastication. PMID- 16690063 TI - Partial-discard strategy for obtaining high purity products using simulated moving bed chromatography. AB - A new "partial-discard" operation strategy was developed to improve the performance, especially purity, achievable in simulated moving bed (SMB) chromatography. This strategy was applied to the four-zone SMB process with two columns per zone for binary separation within the linear range. The "partial discard" strategy significantly enhanced the purity or enrichment when the discard time and discard length were controlled. In addition, the "partial-feed with partial-discard" strategy improved remarkably the extract and raffinate purities at an intermediate feed time compared with the "partial-feed" operation. Adjustments of the discard length and discard time played key roles in achieving the desired product purity in SMB chromatographic performance. PMID- 16690064 TI - Loading, stationary phase, and salt effects during hydrophobic interaction chromatography: alpha-lactalbumin is stabilized at high loadings. AB - Amide hydrogen-deuterium exchange labeling has been used to study the effects of salt and protein loading on alpha-lactalbumin (BLA) stability during hydrophobic interaction chromatography (HIC). Stability in the adsorbed phase increased dramatically with increasing loading, and unfolding was nearly undetectable close to the resin saturation capacity. We also found that a butyl surface destabilized BLA more than a phenyl surface, despite the fact that BLA was bound more strongly on the phenyl surface. These observations have important implications for HIC process design and indicate that in some cases column capacity does not have to be sacrificed to preserve protein stability. PMID- 16690065 TI - Incorporation of a monolithic column into sequential injection system for drug protein binding studies. AB - A sequential injection analysis (SIA) manifold was incorporated with a monolithic strong anion-exchanger disk for on-line drug-protein interaction studies. The antibiotic ciprofloxacin (CF) was selected as a model drug compound. The separation principle was based on the strong retention of bovine serum albumin (BSA) on the monolithic strong anion-exchanger and the liberation/release of the free form of the drug. Elution of the retained BSA was easily achieved by delivering a different mobile phase via the SIA manifold. The type of functional group of the monolithic support, the breakthrough volume and the injected volumes of CF and BSA were studied and optimized. The influence of the variation of incubation time was studied in on-line binding assays. Scatchard plot was employed to obtain the number of binding sites and the equilibrium binding constants. For the off-line study of the CF-BSA binding, two binding classes were determined with constants of (3.16+/-0.21)x10(6)M(-1) and (1.27+/-0.48)x10(4)M( 1) and 6.1+/-1.3 and 17.8+/-3.9 binding sites per class, respectively. In non equilibrium binding experiments the binding rate constant was k(1)=785 M(-1)min( 1). All measurements were monitored with fluorescence (lambda(ext)=300 nm, lambda(em)=460 nm) and spectrophotometric detection (lambda=280 nm). To evaluate the accuracy of the developed method the obtained results were compared versus ultrafiltration experiments and were found in good agreement. PMID- 16690066 TI - Comparison of a non-aqueous capillary electrophoresis method with high performance liquid chromatography for the determination of herbicides and metabolites in water samples. AB - A method of capillary electrophoresis (CE) for the determination of triazine herbicides and some of their main metabolites in water samples has been developed. The proposed CE method includes an off-line solid-phase extraction (SPE) procedure with LiChrolut EN sorbent coupled to a non-aqueous capillary electrophoresis (NACE) separation with UV detection. The target compounds were the chloro-s-triazines simazine, atrazine, propazine; the methyltio-s-triazines ametryn and prometryn and three main derivatives from the atrazine degradation products; namely, deethylatrazine, deethylhydroxyatrazine and deisopropylhydroxyatrazine. The analytical characteristics of the CE method are reported. The repeatability of the method was studied considering the different steps of the method separately in order to determine the contributions of each step to the total variability of the method. The NACE-UV results are compared with those obtained with a high performance liquid chromatography with UV detection (HPLC-UV) method. The same off-line SPE procedure was applied to both techniques. The results obtained show that both methods afford the same results in the analysis of surface and drinking water samples, with a level of significance regarding the F- and t-tests greater than 0.05 in all the cases. The detection limits in surface water samples were in the 0.04-0.32 microg l(-1) and 0.11-1.2 microg l(-1) ranges for the NACE-UV and HPLC-UV methods, respectively. The recoveries (spiked/found) were significantly 100% in all cases. PMID- 16690067 TI - High-performance liquid chromatographic determination of the plasma concentration of K-27, a novel oxime-type cholinesterase reactivator. AB - A simple and reliable HPLC method for the determination of the plasma level of K 27, an oxime type antidote of use in organophosphorus poisoning is presented. Separation was carried out by HPLC using an octyl silica stationary phase and a mobile phase consisting of 93% phosphate buffer (pH 2.6) containing octane sulfate sodium salt, and 7% methanol. Quantitative absorbance was monitored at 286 nm. The calibration curve was linear through the range of 1.25-200 microg/mL, that is well beyond the detected plasma level range of K-27. Limit of quantitation was 5 microg/mL. Intra-day and inter-day precisions of the HPLC determinations gave standard deviations as 0.77 and 2.67%, respectively. Following intramuscular administration of 50 micromol (22.31 mg) K-27 in rats, the maximum of K-27 concentration in plasma was reached at about 15 min giving 186 microg/mL and the t(1/2) was 85 min. K-27 displays initial (from 15 trough 120 min) zero order elimination kinetics. Similar results have been found after intraperitoneal administration. PMID- 16690068 TI - Sensitive gas chromatographic-mass spectrometric method for the determination of phthalate esters, alkylphenols, bisphenol A and their chlorinated derivatives in wastewater samples. AB - Phthalate esters, alkylphenols, bisphenol A and their chlorinated derivatives are the suspected endocrine disrupters or mutagens. These compounds, commonly called endocrine disrupter chemicals (EDCs), are widely used as plastic additives, lacquers, resins, or surfactants and can be usually found in environmental samples, namely wastewaters. An accurate and reproducible gas chromatographic mass spectrometric (GC-MS) method is proposed to measure these compounds in wastewater samples of Granada city (Spain). A solid-phase extraction with LiChrolut RP-18 cartridges was carried out and the elution was performed with a diethyl ether/methanol mixture. After isolation, solvent was removed and a silylation step was carried out using N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA). Phthalate esters and silylated compounds were identified and quantified by GC-MS using a ZB-5 MS column. Bisphenol F was used as a surrogate. Quantification limits found were between 20 ng L(-1) for 4-nonylphenol and 400 ng L(-1) for benzylbutyl phthalate while inter and intra-day variability was under 5% in all cases. Recoveries for spiked samples were over 95% and under 105%. The method was validated using standard addition calibration and recovery assays. PMID- 16690069 TI - Control of silica shell thickness and microporosity of titania-silica core-shell type nanoparticles to depress the photocatalytic activity of titania. AB - Titania is of potential interest as an ultraviolet (UV) radiation blocking material in personal care products because of its excellent UV light absorption properties. Its high photocatalytic activity, however, facilitates the generation of reactive oxygen species, which can oxidize and degrade other ingredients during its formulation, raising safety concerns. Dense coating of titania nanoparticles with a silica layer could help in depression of their photocatalytic activity by disturbing the formation of radicals produced by the reaction of oxygen and/or water with the electron-hole pair. Depression of the high photocatalytic activity of titania necessitates that the silica shell has to be thick, with minimum microporosity. Coating parameters were optimized to attain greater amounts of precipitated silica and thicker shells with lower microporosity, which in turn resulted in great depression of photocatalytic activity. Silica-coated titania nanoparticles were characterized by TEM, XPS, FT IR, EDX, and microporosity measurements. The photocatalytic activity was evaluated for the coated powder to investigate the efficiency of the silica coating as well. PMID- 16690070 TI - Some remarks on the calculation of the pore size distribution function of activated carbons. AB - Different authors investigated the effects of geometric and energetic heterogeneities on adsorption and on carbon characterization methods. In most theoretical studies carbon structure is modeled as parallel infinite graphite walls that form ideal slit-shaped pores of the fixed widths. In the literature there is the lack of systematic studies showing the influence of pore structural and Lennard-Jones (LJ) potential parameters on the pore-size distribution functions. Moreover, the parameters characterizing the properties of the adsorbed phase and the heterogeneity of the adsorbent surface should be taken into account. The Nguyen and Do method with proposed by us ASA algorithm, were utilized for the assessment of the porosity from the series of almost few thousands numerically generated local adsorption isotherms. The values of the mentioned-above parameters are varied over the wide range (ca. +/-20%) of the reference ones. Different types of the theoretical and experimental adsorption isotherms (nitrogen at 77 K) were taken into account as the global ones. They were related to the mechanism of the primary, secondary or mixed micropore filling. The variations in some above-mentioned parameters have significant effects only for PSDs (and for average pore widths) corresponding to the primary micropore filling mechanism. On the other hand, for the process of the secondary micropore filling, the influence of these parameters (without the BET coefficient for adsorption on a "flat" surface, c(s,B)) is rather insignificant. Nevertheless the differences between local and global adsorption isotherms (in the whole range of relative pressures) the absence of micropores having pore half width equal to ca. 1 nm on PSDs was observed for studied adsorbate-adsorbent systems with exceptions of the strictly microporous adsorbents and/or the low values of c(s,B). Comparison of the experimental data with the generated theoretical isosteric enthalpy of adsorption indicates that the phenomenal uptake observed from experiment can be explained in terms of the reasonable solid-fluid interaction parameters. Therefore, we varied the heterogeneity of the adsorbent surface via the strength and the range of the solid-fluid potential and the parameter c(s,B) in order to reproduce the experimental data of enthalpy of adsorption. Note that similar procedure was applied by Wang and Johnson to reproduce some hydrogen adsorption data measured for carbon nanofibres. The analysis of the obtained results shows that the selection of the values of the parameters of the intermolecular interactions and the quantities characterizing the properties of the adsorbed phase and the heterogeneity of the adsorbent walls for molecular simulations should be made with care and the influence of possible errors should be considered. PMID- 16690071 TI - Confirmation of heavy metal ions in used lubricating oil from a passenger car using chelating self-assembled monolayer. AB - In order to prevent engine failure, the oil must be changed before it loses its protective properties. It is necessary to monitor the actual physical and chemical condition of the oil to reliably determine the optimum oil-change interval. Our study focuses on the condition of the lubricating oil in an operated car engine. Shear stress curves and viscosity curves as a function of the shear rate for fresh and used lubricating oil were examined. Metal nitrate was detected in the lubricating oil from the operated car engine through the use of a chelating self-assembled monolayer. PMID- 16690072 TI - Mechanism of oil-in-water emulsification using a water-soluble amphiphilic polymer and lipophilic surfactant. AB - A new O/W (oil-in-water) emulsification system was developed using the amphiphilic polymer HHM-HEC (hydrophobically-hydrophilically modified hydroxyethylcellulose) and a lipophilic surfactant. HHM-HEC was used as a thickener and polymeric surfactant, and the addition of small quantities of various types of nonionic lipophilic surfactant (hydrophilic-lipophilic balance <5) decreased the droplet size of several types of oil due to a lowering of the tension at the water/oil interface. The oil droplets were held by the strong network structure of the aqueous HHM-HEC solution, preserving the O/W phase without inversion. These stable O/W emulsions were prepared without the addition of hydrophilic surfactants and thus show improved water repellency. PMID- 16690073 TI - Surface oxidation of polyethylene using an atmospheric pressure glow discharge with liquid electrolyte cathode. AB - This study investigated the action of an atmospheric pressure air glow discharge (APGD) with aqueous electrolyte cathode onto the surface of polyethylene (PE) films. Distilled water and aqueous solutions of KCl and HCl were utilized as a cathode. The surface properties of PE were characterized by contact angle measurement followed by surface free energy calculation, Fourier transform infrared by attenuated total reflectance (FTIR-ATR), and XPS. After treating the PE surface, we observed OH groups, CO groups in ester, ketone, and carboxyl groups, and CO groups in unsaturated ketones and aldehydes. For a treatment time of 20 min and a discharge current of 40 mA, atomic concentrations of O and N were 12% and 2%, respectively, under distilled water application. Modification processes were able to improve the surface free energy of PE. PMID- 16690074 TI - Behavior of yeast cells in aqueous suspension affected by pulsed electric field. AB - This work discusses pulsed electric fields (PEF) induced effects in treatment of aqueous suspensions of concentrated yeast cells (S. cerevisiae). The PEF treatment was done using pulses of near-rectangular shape, electric field strength was within E=2-5 kV/cm and the total time of treatment was t(PEF)=10(-4) 0.1 s. The concentration of aqueous yeast suspensions was in the interval of C(Y)=0-22 (wt%), where 1% concentration corresponds to the cellular density of 2x10(8) cells/mL. Triton X-100 was used for studying non-ionic surfactant additive effects. The electric current peak value I was measured during each pulse application, and from these data the electrical conductivity sigma was estimated. The PEF-induced damage results in increase of sigma with t(PEF) increasing and attains its saturation level sigma approximately sigma(max) at long time of PEF treatment. The value of sigma(max) reflects the efficiency of damage. The reduced efficiency of damage at suspension volume concentration higher than phi(Y) approximately 32 vol% is explained by the percolation phenomenon in the randomly packed suspension of near-spherical cells. The higher cytoplasmic ions leakage was observed in presence of surfactant. Experiments were carried out in the static and continuous flow treatment chambers in order to reveal the effects of mixing in PEF-treatment efficiency. A noticeable aggregation of the yeast cells was observed in the static flow chamber during the PEF treatment, while aggregation was not so pronounced in the continuous flow chamber. The nature of the enhanced aggregation under the PEF treatment was revealed by the zeta-potential measurements: these data demonstrate different zeta-potential signs for alive and dead cells. The effect of the electric field strength on the PEF-induced extraction of the intracellular components of S. cerevisiae is discussed. PMID- 16690075 TI - Study of the formation of perovskite type lanthanum ruthenates by heating their hydrous precursor. AB - Emanation thermal analysis (ETA), differential thermal analysis (DTA), thermogravimetry (TG), evolved gas analysis with mass spectrometric detection (EGA-MS), and X-ray diffraction (XRD) were used to investigate the formation of perovskite type lanthanum ruthenates on heating their hydroxide precursor in argon from 20 to 1200 degrees C. The co-precipitated lanthanum-ruthenium mixed hydroxide containing a small amount of carbonates was used as a precursor. The mass loss corresponding to the release of water and CO(2) from the precursor was determined by TG and EGA (MS), respectively. The ETA characterized the exposure of sample surface after release of water and CO(2), as well as microstructure development corresponding to the crystallization and structure ordering of LaRuO(3) and La(3.5)Ru(4.0)O(13) perovskite phases. The obtained information on formation of phases and their transformation is useful for optimizing their synthesis protocols for achieving the desired physical properties, and to estimate the thermal stability of these materials to be used as catalysts. PMID- 16690076 TI - Aggregation behavior of polypropylene oxide with electric charges at both ends in aqueous solution. AB - The aggregation behavior of polypropylene oxide (PPO) with positive charges at both ends was investigated in aqueous solution by means of the measurements of solution turbidity, dynamic light-scattering, differential scanning calorimetry, and dye solubilization. The positive charges were produced by protonation of terminal NH(2) groups attached to the polymer composed of 33 PO units. It was found that the aggregation behavior is quite sensitive to temperature. At low temperature, the polymer dissolves in water as a unimer. When temperature is increased, the unimer solution undergoes a phase separation to give a turbid solution. Further increase in temperature produces a transparent micellar solution. The aggregation of the polymer molecules must be induced by the dehydration of PPO chain caused by temperature increase. According to the analysis of heat absorptions associated with the melting of the solid mixture and the phase separation of the unimer solution, it is suggested that approximately 10% dehydration of PPO chain causes the phase separation. The temperature composition phase diagram of aqueous mixture of this polymer was constructed on the basis of turbidity and DSC experiments, which reveals the aggregation behavior of this polymer in aqueous medium as a function of concentration and temperature. PMID- 16690077 TI - Solubilization of pyrene in aqueous micellar solutions of gemini surfactants C12 s-C12.2Br. AB - Solubilization of pyrene in aqueous micellar solutions of quaternary ammonium gemini surfactants C(12)-s-C(12).2Br has been examined by UV spectra and steady fluorescence spectra at 30 degrees C. The results showed that pyrene molecules were incorporated in the palisade layers of the micelles and interacted with the quarterary ammonium head groups through cation-pi interaction, resulting in red shift of the UV absorption spectrum. C(12)-s-C(12)2Br (s=3,4,6) micelles have stronger ability for solubilization of pyrene than conventional surfactant C(12)TABr micelles. With increasing spacer length of C(12)-s-C(12).2Br, the micelles become more adaptable to solubilizing pyrene, which even forms the dimers as in the case of s=3,4,6, while pyrene solubilizes in the micelles (s=2) mainly in single-molecule form due to the more compact structure of the micelle in comparison with that at s>2. PMID- 16690078 TI - Identification of a monoclonal antibody against the leptin receptor that acts as an antagonist and blocks human monocyte and T cell activation. AB - Nutritional status has a major impact on the immune response and this is in part mediated by leptin, a pro-inflammatory cytokine. Preliminary data suggest that antagonism of leptin may offer a therapeutic approach for the treatment of some inflammatory disorders. We have tested monoclonal antibodies (mAbs) to the human leptin receptor (ObR) for antagonist activity using a leptin signalling bioassay. We identified a mAb, 9F8, which demonstrated dose-dependent antagonist activity in the leptin bioassay. Specificity of the mAb for ObR was confirmed using a plate binding assay. The 9F8 mAb displaced leptin binding to human ObR and enzymatically generated Fab fragments of 9F8 retained antagonist activity. Therefore the Fab fragment of 9F8 was cloned and recombinant 9F8 Fab (rFab) was purified from E. coli periplasmic fraction using a C-terminal His tag. Purified 9F8 rFab bound to human ObR and exhibited leptin antagonist activity. In vitro studies demonstrated that the 9F8 mAb inhibited leptin induced TNF-alpha production from human monocytes and anti-CD3 mAb induced proliferation of human T cells in PBMC culture. In conclusion, this study has identified a mAb to the human leptin receptor which inhibits leptin signalling and acts as a leptin antagonist in vitro. PMID- 16690079 TI - Calcium-dependent tetramer formation of S100A8 and S100A9 is essential for biological activity. AB - S100 proteins comprise the largest family of calcium-binding proteins. Members of this family usually form homo- or heterodimers, which may associate to higher order oligomers in a calcium-dependent manner. The heterodimers of S100A8 and S100A9 represent the major calcium-binding proteins in phagocytes. Both proteins regulate migration of these cells via modulation of tubulin polymerization. Calcium binding induces formation of (S100A8/S100A9)2 tetramers. The functional relevance of these higher-order oligomers of S100 proteins, however, is not yet clear. To investigate the importance of higher-order oligomerization for S100 proteins, we created a set of mutations within S100A9 (N69A, E78A, N69A+E78A) destroying the high-affinity C-terminal calcium-binding site (EF-hand II). Mutations in EF-hand II did not interfere with formation of the S100A8/S100A9 heterodimer as demonstrated by yeast two-hybrid experiments and pull-down assays. In contrast, mass spectrometric analysis and density gradient centrifugation revealed that calcium-induced association of (S100A8/S100A9)2 tetramers was strictly dependent on a functional EF-hand II in S100A9. Failure of tetramer formation was associated with a lack of functional activity of S100A8/S100A9 complexes in promoting the formation of microtubules. Thus, our data demonstrate that calcium-dependent formation of (S100A8/S100A9)2 tetramers is an essential prerequisite for biological function. This is the first report showing a functional relevance of calcium-induced higher-order oligomerization in the S100 family. PMID- 16690080 TI - Order of steps in the cytochrome C folding pathway: evidence for a sequential stabilization mechanism. AB - Previous work used hydrogen exchange (HX) experiments in kinetic and equilibrium modes to study the reversible unfolding and refolding of cytochrome c (Cyt c) under native conditions. Accumulated results now show that Cyt c is composed of five individually cooperative folding units, called foldons, which unfold and refold as concerted units in a stepwise pathway sequence. The first three steps of the folding pathway are linear and sequential. The ordering of the last two steps has been unclear because the fast HX of the amino acid residues in these foldons has made measurement difficult. New HX experiments done under slower exchange conditions show that the final two foldons do not unfold and refold in an obligatory sequence. They unfold separately and neither unfolding obligately contains the other, as indicated by their similar unfolding surface exposure and the specific effects of destabilizing and stabilizing mutations, pH change, and oxidation state. These results taken together support a sequential stabilization mechanism in which folding occurs in the native context with prior native-like structure serving to template the stepwise formation of subsequent native-like foldon units. Where the native structure of Cyt c requires sequential folding, in the first three steps, this is found. Where structural determination is ambiguous, in the final two steps, alternative parallel folding is found. PMID- 16690081 TI - Structure of the N-terminal domain of the FOP (FGFR1OP) protein and implications for its dimerization and centrosomal localization. AB - The fibroblast growth factor receptor 1 (FGFR1) oncogene partner, FOP, is a centrosomal protein that is involved in the anchoring of microtubules (MTS) to subcellular structures. The protein was originally discovered as a fusion partner with FGFR1 in oncoproteins that give rise to stem cell myeloproliferative disorders. A subsequent proteomics screen identified FOP as a component of the centrosome. FOP contains a Lis-homology (LisH) motif found in more than 100 eukaryotic proteins. LisH motifs are believed to be involved in microtubule dynamics and organization, cell migration, and chromosome segregation; several of them are associated with genetic diseases. We report here a 1.6A resolution crystal structure of the N-terminal dimerization domain of FOP. The structure comprises an alpha-helical bundle composed of two antiparallel chains, each of them having five alpha-helices. The central part of the dimer contains the LisH domain. We further determined that the FOP LisH domain is part of a longer N terminal segment that is required, albeit not sufficient, for dimerization and centrosomal localization of FOP. PMID- 16690082 TI - Molecular dynamics simulations of the tetracycline-repressor protein: the mechanism of induction. AB - Molecular dynamics simulations on the tetracycline-repressor (TetR) protein, both in the absence of an inducer and complexed with the inducers tetracycline and 5a,6-anhydrotetracycline, show significant differences in the structures and dynamics of the induced and non-induced forms of the protein. Calpha-density difference plots, low-frequency normal vibrations and inter-residue interaction energies all point to a common mechanism of induction. The inducer displaces Asp156 from the magnesium ion in the binding pocket, leading to a short cascade of rearrangements of salt bridges that results in the allosteric change. The increased flexibility of the induced form of the protein is suggested to contribute to the decrease in binding affinity to DNA on induction. PMID- 16690083 TI - Structural recognition between a four-way DNA junction and a resolving enzyme. AB - Resolving enzymes bind highly selectively to four-way DNA junctions, but the mechanism of this structural specificity is poorly understood. In this study, we have explored the role of interactions between the dimeric enzyme and the helical arms of the junction, using junctions with either shortened arms, or circular permutation of arms. We find that DNA-protein contacts in the arms containing the 5' ends of the continuous strands are very important, conferring a significant level of sequence discrimination upon both the choice of conformer and the order of strand cleavage. We have exploited these properties to obtain hydroxyl radical footprinting data on endonuclease I-junction complexes that are not complicated by the presence of alternative conformers, with results that are in good agreement with the arm permutation and shortening experiments. Substitution of phosphate groups at the center of the junction reveals the importance of electrostatic interactions at the point of strand exchange in the complex. Our data show that the form of the complex between endonuclease I and a DNA junction depends on the core of the junction and on interactions with the first six base pairs of the arms containing the 5' ends of the continuous strands. PMID- 16690084 TI - Family history of mood disorder and characteristics of major depressive disorder: a STAR*D (sequenced treatment alternatives to relieve depression) study. AB - INTRODUCTION: Clinicians routinely ask patients with major depressive disorder (MDD) about their family history. It is unknown, however, if patients who report a positive family history differ from those who do not. This study compared the demographic and clinical features of a large cohort of treatment-seeking outpatients with non-psychotic MDD who reported that they did or did not have at least one first-degree relative who had either MDD or bipolar disorder. METHODS: Subjects were recruited for the STAR( *)D multicenter trial. Differences in demographic and clinical features for patients with and without a family history of mood disorders were assessed after correcting for age, sex, race, and ethnicity. RESULTS: Patients with a family history of mood disorder (n=2265; 56.5%) were more frequently women and had an earlier age of onset of depression, as compared to those without such a history (n=1740; 43.5%). No meaningful differences were found in depressive symptoms, severity, recurrence, depressive subtype, or daily function. CONCLUSIONS: Women were twice as likely as men to report a positive family history of mood disorder, and a positive family history was associated with younger age of onset of MDD in the proband. Consistent with prior research, early age of onset appears to define a familial and, by extension, genetic subtype of major depressive disorder. PMID- 16690085 TI - Association between serotonin transporter gene and borderline personality disorder. AB - Borderline personality disorder (BPD) is characterized by a pervasive pattern of instability in regulation of emotion, interpersonal relationships, self-image, and impulse control beginning in early adulthood. BPD affects about 1-2% of the general population and has a high mortality rate as a result of suicide and impulsive behaviour. The serotonin transporter gene (5-HTT) is considered as a candidate gene for BPD as multiple lines of evidence have suggested that it plays an important role in suicide, impulsive behaviour, and emotional liability. To test for an association between 5-HTT and BPD, we genotyped three common polymorphisms: the serotonin transporter linked promoter region (5-HTTLPR); a variable number of tandem repeat (VNTR) in intron 2, and a single nucleotide variant (A/G) within the LPR region. Eighty-nine Caucasian patients with BPD and 269 Caucasian healthy controls were analyzed. The program UNPHASED was used to compare allele and haplotype frequencies between cases and controls. Significant differences in allele frequencies of the VNTR marker (p=0.012) and haplotype frequencies (p=0.002) between patients and controls were found. Compared with healthy controls, patients with BPD showed higher frequencies of the 10 repeat of the VNTR marker and the S-10 haplotype, and lower 12 repeat and L(A)-12 haplotype. Our results suggest that the serotonin transporter gene may play a role in the aetiology of borderline personality disorder. PMID- 16690086 TI - Driving performance while using cell phones: an observational study. AB - INTRODUCTION: Through spontaneous driving observations, this study sought to examine the impact of using a hands-free cell phone while driving on speed and safe gap keeping behaviors. The study also examined the association between the measure of disturbance created by using a cell phone and the driver's awareness of the disturbance. METHOD: Twenty-three male adults were observed while driving for an hour and a half each; drivers were unaware of being observed. During the session, each of the participants received a phone call, initiated by an associate of the observer. The experiment was divided into two periods during which the experimental parameters were monitored: 10 minutes during conversation on a cell phone and 10 minutes of non-conversation on a hands-free cell phone. After the experiment, the driver was questioned concerning the extent to which his/her driving was disturbed by the cell phone conversation. RESULTS: T-test for matched samples revealed that the gaps between the drivers' cars and those in front of them diminished when drivers were engaged in the cell phone conversations. Repeated measures ANOVA revealed that drivers that had short conversations did not change their speed, while drivers who were engaged in long (over 16 minutes) conversations drove faster. No effect of drivers' awareness toward cell phone-related disturbance on actual driving behavior monitored in the present study was found. PMID- 16690087 TI - Prenatal nicotine exposure alters the nicotinic receptor subtypes that modulate excitation of parasympathetic cardiac neurons in the nucleus ambiguus from primarily alpha3beta2 and/or alpha6betaX to alpha3beta4. AB - Nicotinic receptors play an essential role in central cardiorespiratory function, however, the types of nicotinic receptors responsible for activating cardiac vagal neurons in the nucleus ambiguus that control heart rate are unknown. This study tests whether alpha-conotoxin MII and alpha-conotoxin AuIB sensitive nicotinic receptors are involved in augmentation of glutamatergic neurotransmission and changes in holding current in cardiac vagal neurons, and whether exposure to nicotine in the prenatal period alters these responses. The nicotinic agonist cytisine significantly increased the holding current and amplitude of glutamatergic mEPSCs. In unexposed animals alpha-conotoxin MII (100nM) significantly reduced the increase in mEPSC amplitude and change in holding current evoked by cytisine. However, in animals prenatally exposed to nicotine, alpha-conotoxin MII blunted but did not block the increase in mEPSC amplitude but blocked the increase in holding current evoked by cytisine. In unexposed animals, alpha-conotoxin AuIB (10microM) blocked the cytisine evoked increase in mEPSC amplitude and inhibited but did not abolish the increase in holding current. In contrast, in animals exposed to nicotine, alpha-conotoxin AuIB blunted the increase in mEPSC amplitude, and completely abolished the cytisine evoked increase in holding current. These data demonstrate that the prenatal nicotine exposure alters the nicotinic receptors involved in excitation of cardiac vagal neurons. PMID- 16690088 TI - Postural control after a night without sleep. AB - The present study analysed the efficiency of postural control after 12 h of nocturnal forced wakefulness using Romberg's test comprising 1 min of recording with eyes-open and 1 min of recording with eyes-closed, with a 1 min break between the two sessions. Our aim was to see if the decreased postural control efficiency after a sleepless night was unspecific (in both eyes-closed and eyes open conditions) or selective (in only one of the conditions). A total of 55 students spent a whole night awake at our laboratory and were tested at 22:00 and 08:00 h. In general, the results showed that postural sway increased, performing the recording from eyes-open to eyes-closed condition. The statokinesigram length (SL or efficiency of the postural system) increased after the sleepless night, while in eyes-open condition, the length in function of surface (LFS or accuracy of postural control) and Romberg's index (or contribution of vision to maintain posture) significantly decreased. This could indicate that after a night without sleep, there is a slower elaboration of visual inputs in the postural control process. On the basis of these results, the effects of sleep deprivation on cognitive performance were considered from a neuropsychological point of view. PMID- 16690089 TI - A dissociation between visual and auditory hemi-inattention: Evidence from temporal order judgements. AB - Patients with right hemisphere brain lesions often suffer from deficits in spatial attention that can be manifested in different sensory modalities. It has recently been claimed that a relationship (i.e., association) could exist between symptoms of hemi-inattention in different modalities, based on correlations between the results of visual and auditory clinical tests of neglect or extinction. However, it should be noted that the visual and auditory tasks varied greatly both in response type and level of sensitivity. Here, we have examined cross-modal associations in spatial attention deficits using a temporal order judgment task (TOJ) in which patients were required to identify which of two visual or auditory objects had appeared first. When compared to age and education matched control participants, the patients needed, on average, the contralesional stimulus to lead the ipsilesional stimulus to achieve the point of subjective simultaneity (PSS). No association between the degree of visual and auditory hemi inattention was observed amongst the patients, suggesting that there is a certain degree of independence between the mechanisms subserving spatial attention across sensory modalities. PMID- 16690090 TI - D2 dopamine receptor blockade prevents cognitive effects of Ang IV and des-Phe6 Ang IV. AB - Angiotensins, especially angiotensin IV (Ang IV), have recently been found to be potent cognitive enhancers in rodents. However, the precise mechanisms of their memory improving effects remain unknown. In this study we tested the hypothesis that D2 dopamine receptors at least partially mediate cognitive effects of Ang IV and its derivative des-Phe6 Ang IV. Namely, the well known cognitive effects of both peptides [facilitation of a conditioned avoidance responses (CARs) acquisition, increase of a passive avoidance behavior (PAB) retrieval, and improvement of object recognition] were evaluated in rats either pretreated or not with a selective D2 dopamine receptor antagonist remoxipride {(S)-(-)-3-Bromo N-[(1-ethyl-2-pyrrolidinylOmethyl]2,6-dimethoxybenzamide hydrochloride}. To control for the unspecific motor and emotional effects of our treatments that could confound results of the memory tests we used respectively, 'open' field and elevated 'plus' maze tests. Ang IV as well as des-Phe6 Ang IV remarkably improved learning of CARs, recall of PAB and recognition of the previously seen objects. D2 receptors blockade by remoxipride abolished all these effects of both peptides. In the elevated 'plus' maze remoxipride abolished anxiogenic effects of both Ang IV and des-Phe6 Ang IV. Also, the drug followed by Ang IV decreased number of crossings and by des-Phe6 Ang IV number of crossings and rearings. The results point to importance of the functional D2 dopamine receptors in cognitive effects of Ang IV and its naturally occurring product devoid of C-terminal Phe6. PMID- 16690091 TI - Ethological analysis of scopolamine treatment or pretreatment in morphine dependent rats. AB - Although scopolamine is currently used to treat morphine addiction in humans, its extensive actions on behaviors have not been systematically analyzed yet, and the underlying mechanisms of its effects still remain ambiguous. The present study was carried out to clarify the possible mechanisms by evaluating the effects of scopolamine pretreatment and treatment on naloxone-precipitated withdrawal signs and some of other general behaviors in morphine dependent rats. Our results showed that scopolamine pretreatment and treatment attenuated naloxone precipitated withdrawal signs including jumping, writhing posture, weight loss, genital grooming, teeth-chattering, ptosis, diarrhea and irritability, except for wet dog shakes, while general behaviors such as water intake, urine volume and morphine excretion in urine were increased. Our findings suggest that scopolamine has significant actions in the treatment of opiate addiction, which might result from increasing morphine excretion from urine. PMID- 16690092 TI - Labdane diterpenes from Leonurus japonicus leaves. AB - Three labdane diterpenes 15,16-epoxy-6-hydroxylabda-5,8,13(16),14-tretraen-7-one (leojaponin), (9alpha,13S);15,16-diepoxy-7beta-hydroxylabd-14-en-6-one (13-epi preleoheterin), and (9alpha,13R);15,16-diepoxy-6beta-hydroxylabd-14-en-7-one (iso preleoheterin) were isolated from the leaves of Leonurus japonicus, in addition to the previously reported preleoheterin. The structure elucidations were made based on analysis of their spectroscopic data. PMID- 16690093 TI - HIV status disclosure and partner discordance: a public health dilemma. PMID- 16690094 TI - Prevalence of Plasmodium falciparum cytochrome b gene mutations in isolates imported from Africa, and implications for atovaquone resistance. AB - The atovaquone resistance of malaria parasites correlates with mutations in the cytochrome b gene. We sequenced the Plasmodium falciparum cytochrome b gene of 135 African isolates. Our data showed a high mutation rate (8.9%); however, the risk of emergence spreading of atovaquone-resistant P. falciparum strains could be limited. PMID- 16690095 TI - Ultrasonic density measurement cell design and simulation of non-ideal effects. AB - This paper presents a theoretical analysis of a density measurement cell using an unidimensional model composed by acoustic and electroacoustic transmission lines in order to simulate non-ideal effects. The model is implemented using matrix operations, and is used to design the cell considering its geometry, materials used in sensor assembly, range of liquid sample properties and signal analysis techniques. The sensor performance in non-ideal conditions is studied, considering the thicknesses of adhesive and metallization layers, and the effect of residue of liquid sample which can impregnate on the sample chamber surfaces. These layers are taken into account in the model, and their effects are compensated to reduce the error on density measurement. The results show the contribution of residue layer thickness to density error and its behavior when two signal analysis methods are used. PMID- 16690096 TI - Long-lived memory T lymphocyte responses against SARS coronavirus nucleocapsid protein in SARS-recovered patients. AB - The nucleocapsid (N) protein is a structural component of severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) and can induce antibody responses in SARS patients during infection. However, it is not known whether SARS-CoV N protein can induce a long persistence of memory T-cell response in human. In this study, we found that peripheral blood mononuclear cells (PBMCs) from fully recovered SARS individuals rapidly produced IFN-gamma and IL-2 following stimulation with a pool of overlapping peptides that cover the entire N protein sequence. The N-specific IFN-gamma(+)CD4(+) T cells were mainly composed of CD45RA(-)CCR7(+)CD62L(-) cells, whereas IFN-gamma(+)CD8(+) memory T cells were mostly contained within CD45RA(+)CCR7(-)CD62L(-) cell population. Epitope mapping study indicated that a cluster of overlapping peptides located in the C-terminal region (amino acids [aa] 331 to 362) of N protein contained at least two different T-cell epitopes. The results indicated that human memory T-cell responses specific for SARS-CoV N protein could persist for 2 years in the absence of antigen, which would be a valuable for the design of effective vaccines against SARS-CoV and for basic studies of human T-cell memory. PMID- 16690097 TI - The immunogenicity and efficacy against H5N1 challenge of reverse genetics derived H5N3 influenza vaccine in ducks and chickens. AB - H5N1 avian influenza viruses are continuing to spread in waterfowl in Eurasia and to threaten the health of avian and mammalian species. The possibility that highly pathogenic (HP) H5N1 avian influenza is now endemic in both domestic and migratory birds in Eurasia makes it unlikely that culling alone will control H5N1 influenza. Because ducks are not uniformly killed by HP H5N1 viruses, they are considered a major contributor to virus spread. Here, we describe a reverse genetics-derived high-growth H5N3 strain containing the modified H5 of A/chicken/Vietnam/C58/04, the N3 of A/duck/Germany/1215/73, and the internal genes of A/PR/8/34. One or two doses of inactivated oil emulsion vaccine containing 0.015 to 1.2 microg of HA protein provide highly efficacious protection against lethal H5N1 challenge in ducks; only the two dose regimen has so far been tested in chickens with high protective efficacy. PMID- 16690098 TI - Perceptual learning with spatial uncertainties. AB - In perceptual learning, stimuli are usually assumed to be presented to a constant retinal location during training. However, due to tremor, drift, and microsaccades of the eyes, the same stimulus covers different retinal positions on sequential trials. Because of these variations the mathematical decision problem changes from linear to non-linear (). This non-linearity implies three predictions. First, varying the spatial position of a stimulus within a moderate range does not deteriorate perceptual learning. Second, improvement for one stimulus variant can yield negative transfer to other variants. Third, interleaved training with two stimulus variants yields no or strongly diminished learning. Using a bisection task, we found psychophysical evidence for the first and last prediction. However, no negative transfer was found as opposed to the second prediction. PMID- 16690099 TI - Color discrimination in carriers of color deficiency. AB - Carriers of X-linked color vision deficiencies have previously been reported to exhibit mild abnormalities of color matching and discrimination. In a sample of 55 carriers of protan and deutan deficiencies and 55 age-matched normal controls, we measured chromatic discrimination along a red-green axis. We found that discrimination was impaired in the case of carriers of deutan deficiencies (which affect the middle-wave-sensitive cones of the retina), but was normal in the case of carriers of protan deficiencies (which affect the long-wave-sensitive cones). We argue that this result can be explained by the difference in the relative numbers of middle- and long-wave cones in heterozygous retinae: the imbalance of the two cone types is predicted to be much greater in the case of the deutan heterozygote than in the case of the protan heterozygote. In future studies it will be necessary to consider separately the two types of heterozygote. PMID- 16690100 TI - Sensitive and meaningful measures of bacterial metabolic activity using NADH fluorescence. AB - Successful biological wastewater treatment depends on bacterial metabolic activity. Commercial fluorimeters are designed to monitor this activity using the native fluorescence of Nicotinamide Adenine Dinucleotide [NADH]. However, fluorescence measurements in wastewater treatment plants remain scarce due to difficulties with interpreting fluorescence data. This paper shows that fluorescence probe measurements taken from wastewater do not represent bacterial cell metabolic activity because intracellular NADH is likely swamped by the stable extracellular NADH fraction. Thus, a simple filtration/extraction/centrifugation method was developed to collect the bacterial cells, extract the intracellular NADH using heat treatment in Tris buffer and collect the purified intracellular NADH fraction. NADH standards were used to quantify NADH from the unknown wastewater samples where limits of detection were between 1 nmol mL(-1) and 0.35 micromol mL(-1). Fluorescence of [NADH] greater than 0.35 micromol mL(-1) was self-quenched. At high pH's NADH was stable outside the cell. NADH was stable at neutral and basic pH ranges of pH 7 to 11, but declined proportionally below a pH of 7. Since commercially available fluorescence probes used for measuring NADH are more likely detecting extracellular NADH, separating bacterial cells from water samples followed by NADH extraction was essential to distinguish intracellular and extracellular [NADH]. Here we have proposed three simple steps to meaningful measures of bacterial metabolic activity based on the autofluorescence of NADH. The three simple steps to getting it right are Future development of an on-line monitoring system based on these three steps is achievable with a little ingenuity. PMID- 16690101 TI - Total chemical synthesis of the B1 domain of protein L from Peptostreptococcus magnus. AB - Reported here is a native chemical ligation strategy for the total chemical synthesis of the B1 domain of protein L. A synthetic construct of this 76 amino acid protein domain was prepared by the chemoselective ligation of two unprotected polypeptide fragments, one containing an N-terminal cysteine residue and one containing a C-terminal thioester moiety. The polypeptide fragments utilized in the ligation reaction were readily prepared by stepwise solid phase peptide synthesis (SPPS) methods for Boc-chemistry. The milligram quantities of protein required for conventional biophysical studies were readily accessible using the synthetic protocol described here. The folding properties of the synthetic protein L construct were also determined and found to be very similar to those of a similar wild-type protein L constructs prepared by recombinant-DNA methods. This work facilitates future unnatural amino acid mutagenesis experiments on this model protein system to further dissect the molecular basis of its folding and stability. PMID- 16690102 TI - Photochemical reduction and reoxidation of aqueous mercuric chloride in the presence of ferrioxalate and air. AB - In this study, ferric oxalate is used to represent the photosensitive Fe(III) complexes as well as the diacid compounds which are at significant concentrations in cloud and rain droplets. Because of the common carboxylate functional group; ferric oxalate is also used as a model to represent humic substances found in natural water. UVA irradiation of aqueous acidic mercuric chloride (pH 1-4) in the presence of an excess of ferrioxalate results in partial reduction of the mercuric ion to elemental mercury. The pseudo-first-order rate constant "kobs" for the photoreduction reaction is pH-dependent as is the yield of residual Hg(II). When exposed to visible irradiation the rate is about 10 times slower and no reaction was observed in the dark. The inferred mechanism of photoreduction involves the reaction of Hg(II) with a secondary photoproduct, the strongly reducing radical anion CO2-*. In the presence of dissolved oxygen, competition for CO2-* between Hg(II) and O2 reduces the rate and efficiency of mercuric ion reduction. The O2-*/HO2 products do not reduce Hg(II). On the contrary, their disproportionation leads to the formation of H2O2 which causes a re-oxidation of Hg(0) at pH values of 2 cm, neuroendocrine tumor type, tumor-involved resection margins, or positive pelvic lymph nodes). Thus, 100 patients were treated by RVT according to protocol. With 4 recurrences in a sample size of 100 patients, an upper limit of the 95% confidence interval (including continuity correction) of 10.5% was calculated. Recruitment had to be stopped if five or more recurrences occurred. RESULTS: The median follow-up time was 29 (1-128) months. Three (3%) recurrences occurred in 100 patients treated with RVT according to protocol. Thus, the upper confidence limit was 9.2%. The projected 5-year recurrence-free and overall survival rates were 97% and 98%. The average duration of surgery was 253 (115 402) min. Perioperative complications were: postoperative bleeding, embolism of the external iliac artery, retroperitoneal lymphocele, or paralytic ileus in one patient, respectively. CONCLUSIONS: RVT combined with laparoscopic pelvic and parametric lymphadenectomy for treatment of patients with early stage cervical cancer < or =2 cm results in a recurrence-free survival of more than 90.8%. PMID- 16690105 TI - Determination of the mechanism of gemcitabine modulation of cisplatin drug resistance in panel of human endometrial cancer cell lines. AB - OBJECTIVES: The primary objective of this study was to determine the mechanism(s) of cisplatin drug resistance in endometrial cancer cell lines. To evaluate the mechanism that gemcitabine modulates cisplatin drug resistance in endometrial cancer cell lines. METHODS: Combination treatment was completed in panel of four human endometrial cancer cell lines. Growth inhibition assays were conducted in each cell line evaluating combinations of the Ic25, Ic50, and Ic90 to determine optimal dosing for the combination of gemcitabine plus cisplatin. Evaluation of the correlative biological targets for modulation of platinum drug resistance was completed by the respective immunohistochemistry assays. RESULTS: Downregulation of glutathione-S-transferase (GST) activity by 11% to 100% was observed with an associated 78.6% to 100% decrease in intracellular glutathione (GSH) concentrations. In the gemcitabine plus cisplatin treatment arm compared to either alone, there was also downregulation of MSH2, p53, and ERCC1 expression. No changes observed in the pro-apoptotic proteins, BAX or BAD, expression, AKT activation, or MDR1/PGP expression regardless of treatment with combination of gemcitabine plus cisplatin or either agent alone. CONCLUSIONS: There is likely more than one mechanism contributing to the increase synergistic in vitro platinum-resistant cell lines and increase clinical activity that has been observed in patients with platinum-resistant tumors. In this in vitro study, we determined the downregulation of intracellular GST activity and GSH concentration were the predominant mechanisms involved in the modulation of platinum resistance. Downregulation of MSH2, p53 and ERCC1 expression may also contribute to increase cytotoxic activity compared to cisplatin alone. PMID- 16690106 TI - Estrogen-receptor-dependent regulation of telomerase activity in human endometrial cancer cell lines. AB - OBJECTIVES: Given that prolonged exposure to unopposed estrogen is associated with endometrial cancer development and that the promoter region of the catalytic subunit of the telomerase enzyme, hTERT, contains putative estrogen response elements (EREs), we postulated that estrogen-receptor (ER)-mediated induction of telomerase activity may play an important role in endometrial carcinogenesis. METHODS: ER-positive and ER-negative endometrial cancer cell lines were used. ER alpha expression was reconstituted in ER-negative cell lines by transient transfection. Telomerase activity was assayed using a PCR-based telomeric repeat amplification protocol (TRAP) after exposure to estradiol (E2). hTERT mRNA expression was assessed by real-time RT-PCR. Gel shift assays using oligonucleotide probes encoding each ERE and transient expression assays using luciferase reporter plasmids containing varying lengths of the 5' promoter region of the hTERT gene were performed. RESULTS: E2 induced both hTERT gene transcription and telomerase activity in the ER-positive cell lines, but not in the ER-negative cell lines. Transfection of ER alpha into ER-negative cell lines restored E2-induced hTERT gene transcription and telomerase activity. Gel shift assays revealed two EREs in the hTERT promoter that specifically bind to ER alpha. Luciferase assays demonstrated that at least the proximal ERE is responsible for transcriptional activation by ligand-stimulated ER alpha. CONCLUSIONS: Telomerase activity and hTERT mRNA were increased in response to estrogen in an ER alpha-dependent fashion in endometrial cancer cells. Binding of complexed estrogen with ER alpha to the EREs found within the hTERT promoter suggests a possible mechanism for telomerase induction that may facilitate the malignant transformation of hormone-dependent endometrial cells. PMID- 16690107 TI - Body weight and composition changes in ovarian cancer patients during adjuvant chemotherapy. AB - OBJECTIVE: To prospectively examine body weight changes in women with newly diagnosed ovarian cancer receiving surgery and adjuvant chemotherapy. Body composition was examined in a subset of these women. METHODS: Body weight (BW) and body composition, using bioelectrical impedance (RJL Systems Inc.), were prospectively measured pre- and post-operatively, and at 3, 6, and 12 months. RESULTS: Mean age of 42 women was 59 years and did not differ by stage of disease. Nine women with early stage disease did not receive adjuvant chemotherapy. Mean BW of 33 patients receiving chemotherapy decreased from the pre- to post-operative visit and then returned to baseline levels by 12 months (F = 8.70, P = 0.003). Nine patients who did not receive chemotherapy demonstrated a similar pattern (F = 7.0, P = 0.002). Women receiving chemotherapy with stage I/II cancer had a 2.8 +/- 2.0 kg weight gain over the year, and women with stage III/IV cancer had a 1.5 +/- 1.5 kg weight loss (t = 1.72, P = 0.096). A subset of women with stage I/II (n = 6) and stage III/IV (n = 6) ovarian cancer receiving chemotherapy had body composition measured at three time points. Absolute body fat changes paralleled changes in BW (F = 9.95, P = 0.002). CONCLUSIONS: Our study is the first prospective evaluation of body weight and composition in women undergoing surgery and chemotherapy for ovarian cancer. These results demonstrate that women undergoing surgery for ovarian cancer lost weight following surgery and regained it slowly over the following year. Further investigations of weight changes during adjuvant chemotherapy are indicated to assess potential changes in different stages of disease. PMID- 16690108 TI - Exclusion of elderly persons from health-risk behavior clinical trials. AB - BACKGROUND: It has been found that the elderly population is underrepresented in clinical trials aimed at disease treatment. It is unknown to what extent this group is also excluded from clinical trials aimed at reducing health-risk behaviors. METHOD: We systematically reviewed clinical trials targeting the leading health-risk behaviors, as identified by Healthy People 2010: tobacco use, overweight/obesity, physical inactivity, substance abuse, and irresponsible sexual behavior. Using MEDLINE, we identified clinical trials published in the five most cited medical journals between January 1990 and May 2004. RESULTS: The majority (53%) of the 198 health-risk behavior clinical trials excluded persons over the age of 65; the exclusion percentage increased to 72% for those over the age of 75. The exclusion of elderly persons in these five medical journals did not decline over the 14 years studied. This age exclusion pattern was not explained by the intervention's intrusiveness or whether illness was an exclusion criterion. The trials that included those over the age of 65 were more likely to report a significant finding in the predicted direction. CONCLUSIONS: Although elderly persons are most at risk for diseases linked to lifestyle behavior, such as heart disease, they were excluded from the majority of health-risk behavior clinical trials in five major medical journals. PMID- 16690109 TI - Superovulatory response of Sistani cattle to three different doses of FSH during winter and summer. AB - Objective of the present study was to investigate the effect of season and dose of FSH on superovulatory responses in Iranian Bos indicus beef cattle (Sistani). Cyclic cows, in summer (n=16) and winter (n=16), were assigned randomly to three dose-treatment groups of 120 (n=10), 160 (n=12) and 200 (n=10) total mg of Folltropin-V with injections given twice daily for 4 days in decreasing doses. Estrous cycles were synchronized with two prostaglandin F2alpha injections given 14 days apart. From day 5 after the ensuing cycle, daily ovarian ultrasonography was conducted to determine emergence of the second follicular wave at which time superovulation was initiated. Relative humidity, environmental and rectal temperatures were measured at 08:00, 14:00 and 20:00 h for the 3 days before and 2 days after the estrus of superovulation. Non-surgical embryo recovery was performed on day 7 after estrus. The effects of season, dose, time of estrous expression and all two-way interactions were evaluated on superovulatory responses: total numbers of CL, unovulated follicles (10 mm), ova/embryo, transferable and non-transferable embryos. Season (summer or winter), doses of Folltropin-V (120, 160 or 200 mg NIH) and time of estrous expression (08:00, 14:00 or 20:00 h) did not affect the number of transferable embryos (3.1+/-0.58). When superovulatory estrus was detected at 08:00, a FSH dose effect was detected with the greatest numbers of CL (12.2+/-0.87) and total ova/embryos (12.2+/-1.46) occurring with 200 mg FSH (dosextime of estrous expression; P<0.01). PMID- 16690111 TI - The effect of word length on lexical decision in dyslexic and normal reading children. AB - In the present study, the effect of word length on lexical decision in dyslexic and normal reading children was investigated. Dyslexics of 10-years old, chronological age controls, and reading age controls read words and pseudowords consisting of 3 to 6 letters in a lexical decision task. Length effects were much stronger in dyslexics and reading age controls than in chronological age controls. These results support the contention that dyslexics continue to rely on a predominantly sub-lexical reading procedure, whereas for normal readers the contribution of a lexical reading procedure increases. The relevance of the findings for current computational models of reading is discussed. PMID- 16690112 TI - Interhemispheric communication involving multiple tasks: A study of children with dyslexia. AB - The study investigated whether inefficient interhemispheric communication is involved in developmental dyslexia using multiple tasks. A finger localization task, rhyming judgment task, primed lexical decision task, and a visual half field presentation paradigm were used. Nineteen dyslexic children (mean age = 13.1 years) were compared with 26 chronological age-matched normal children. Although the dyslexic group demonstrated significantly slower and less accurate performance in all three tasks, there was no significant group difference in term of interhemispheric communication. However, priming effects demonstrated by the dyslexic group (p < .05) further indicate that their reading problems may stem from the word retrieval process from the long term memory. PMID- 16690113 TI - Technique sensitivity associated with air-drying of HEMA-free, single-bottle, one step self-etch adhesives. AB - OBJECTIVES: This study examined the effect of air-drying on bond strength to dentin and nanoleakage in resin-dentin interfaces using HEMA-free one-step self etch adhesives. METHODS: Twelve teeth had their occlusal dentin exposed and split in half. In the group of G-BOND (GB), the adhesive was dried gently or vigorously on each slit half tooth, and then light-cured. In the group of the experimental adhesive (EX), three coats of adhesive were dried using a single air-drying process prior to light-curing or dried after each of three coats of adhesive before light-curing. The split specimens were built-up with resin composite and then subjected to microtensile bond strength testing and TEM examination. RESULTS: GB showed significantly higher bond strength after vigorous drying than after gentle drying. Three teeth of from EX group showed higher bond strength after triple drying than after single drying, while two teeth showed no significant difference. Droplets were revealed in the adhesive layer after gentle drying with GB, and silver deposits existed along the interfaces in all groups. SIGNIFICANCE: The bonding quality to dentin may be improved by using an accelerated air-drying technique; although it is impossible to remove intrinsic dentin moisture that is drawn upwards from dentin tubules. PMID- 16690114 TI - Temperature-dependent effects of cadmium on mitochondrial and whole-organism bioenergetics of oysters (Crassostrea virginica). AB - Intertidal mollusks are exposed to multiple stressors in estuaries, including temperature and trace metals such as cadmium, which may interactively affect their physiology. We have studied the combined effects of temperature and cadmium stress on metabolism of oysters at the whole animal and mitochondrial levels. In vivo exposure to 50 microg L(-1) Cd led to a significant increase in basal metabolic rate (BMR) in 20 degrees C-acclimated but not in 28 degrees C acclimated oysters. Cadmium exposure resulted in a fast decrease in mitochondrial capacity to synthesize ATP in 28 degrees C-acclimated but not 20 degrees C acclimated oysters indicating that mitochondria may be functioning closer to their capacity limits in the former group. This agrees with elevated mortality in Cd-exposed oysters at 28 degrees C but not 20 degrees C. In general, elevated temperature increased sensitivity of oysters to cadmium at mitochondrial and whole-organism levels suggesting that oyster populations may become more susceptible to trace metal pollution during seasonal warming and/or global climate change. PMID- 16690115 TI - Exploring the use of caged Nucella lapillus to monitor changes to TBT hotspot areas: a trial in the River Tyne estuary (UK). AB - Caged dogwhelks (Nucella lapillus) and mussels (Mytilus edulis) were placed in situ at sites along the River Tyne (UK) in order to investigate the potential for monitoring any change in tributyltin (TBT) mediated effects. After 6 months the development of imposex was investigated in the dogwhelks and concentrations of TBT were determined in the whole tissues of both species. Sites within the estuary showed a high degree of imposex, whilst those at sites North and South of the estuary mouth exhibited significantly lower induction. Concentrations of TBT measured in the dogwhelk and mussel tissues showed the same pattern. The success of the caging technique, occurrence of imposex at other UK sites and the need for further monitoring at TBT impacted locations in the light of current international controls on the use of TBT-based antifouling paints on sea-going vessels are discussed. PMID- 16690116 TI - PCB congener-specific disruption of reproductive neuroendocrine function in Atlantic croaker. AB - Exposure of Atlantic croaker to Aroclor 1254 has been shown to impair reproductive neuroendocrine function in this species. In addition, we have identified hypothalamic tryptophan hydroxylase (TPH), the rate-limiting enzyme in serotonin synthesis, as a target of PCB neuroendocrine toxicity. A previous study in rats has implicated di-ortho-substituted non-coplanar PCB congeners in the inhibition of a similar enzyme, tyrosine hydroxylase, which is the rate-limiting enzyme in dopamine synthesis. Therefore, the present study was designed to investigate whether di-ortho-substituted congeners (PCB 47, PCB 153) or a coplanar congener (PCB 77) present in Aroclor 1254 could be responsible for the reproductive impairment observed in croaker exposed to the PCB mixture. Fish were exposed to PCB 47 and PCB 153 in the diet (0, 0.2 and 1.0 mg/kg body weight/day) for 30 days and to PCB 77 (0.01 and 0.1 mg/kg body weight/day) for 15 days. Neither PCB 47 nor PCB 153 altered hypothalamic TPH activity or gonadal growth at doses similar to the effective doses of the Aroclor 1254 mixture. Therefore, these ortho-substituted PCB congeners known to be neurotoxic in mammalian systems are unlikely to contribute to Aroclor 1254-induced reproductive neuroendocrine disruption in croaker. In contrast, PCB 77 significantly inhibited hypothalamic TPH activity and gonadal growth at doses much lower than the effective doses of Aroclor 1254. The results provide the first evidence for PCB congener-specific disruption of reproductive neuroendocrine function in a vertebrate species. PMID- 16690117 TI - Vitellogenin gene expression as a biomarker of endocrine disruption in the invertebrate, Mytilus edulis. AB - Vitellogenin levels are often used as a biomarker of endocrine disruption in fish. For invertebrates, there is a general lack of knowledge regarding endocrine regulation and, consequently, there are few direct biomarkers of endocrine disruption in such species. This study focuses on the marine mussel Mytilus edulis, which is often employed in biomonitoring studies. A partial vitellogenin mRNA has been isolated and gene expression quantified using real-time PCR. PMID- 16690118 TI - Metallothionein gene expression in zebrafish embryo-larvae and ZFL cell-line exposed to heavy metal ions. AB - The aim of this study is to investigate the induction of zebrafish metallothionein (zMT) gene expression following the administration of different metal ions using in vivo and in vitro models. The zebrafish embryo-larvae were used for the in vivo study, and MT gene expression was studied during the development from fertilization (8hpf) to embryo-larval stage using real-time PCR. The LC50 values and zMT mRNA levels were also measured in embryo-larvae exposed to various metal ions. The general trend of 24 h LC50 values as determined is Cu2+ < Hg2+ < Cd2+ << Zn2+. However, Hg2+ was found to be the most potent metal inducer with the highest level of zMT mRNA induction (40-50 folds) in 8hpf embryo larvae, followed by Cd2+ (approximately 20 folds); Cu2+ and Zn2+ only gave approximately 5 fold of induction. In the in vitro study of ZFL cell-line, Cd2+ is the most potent inducer of zMT mRNA (up to 250 folds), Cu2+ and Zn2+ gave similar potency of approximately 50-100 folds, and Hg2+ gave approximately 40-50 folds of zMT mRNA levels over the control group. PMID- 16690119 TI - Alterations in hepatic gene expression by trivalent chromium in Fundulus heteroclitus. AB - Cr(III) is the dominant toxicant at some Superfund sites within the United States and therefore we are interested in its effects. Cr(III)s mechanisms are not well studied or understood because of its low bioavailability. We have attempted to characterize the effects of Cr(III) on gene expression in Fundulus heteroclitus (mummichog) liver. The NOEC and LOEC were determined at 32 and 64 mg/L, respectively, by measuring growth and mortality after exposing juveniles for 30 days. Secondary adult male exposures were performed at 32 mg/L, livers excised, and RNA extracted. Arrays were probed with cDNA from untreated or Cr(III)-exposed adult fish and gene expression was quantified. Cr(III) at 32 mg/L altered the expression of five genes, including GSTtau, GSTalpha, and ALDH4. Ultimately, we anticipate using this gene expression information to ascertain whether Cr(III) is bioavailable at potentially adverse concentrations in contaminated sites. PMID- 16690120 TI - Crystal structure characterization of nautilus shell at different length scales. AB - In this work, we studied the shell structure of the Nautilus pompilius Linnaeus by using analytical techniques of scanning (SEM) and transmission electron microscopy (TEM) and X-ray diffraction. The main objective of this study is the structural characterization of Nautilus shell at different length levels, from micron to nano-scale. The results were also used to try to determine the shell structure mechanism of formation. The information obtained in this work will place our particular knowledge a closer step to understand how self-assembly works in nature, and will increase the opportunities of using this information in the future synthesis of new advanced materials. PMID- 16690121 TI - Osteoinduction with highly purified beta-tricalcium phosphate in dog dorsal muscles and the proliferation of osteoclasts before heterotopic bone formation. AB - The aim of the study was to examine the chronological histology of osteoinduction of highly purified beta-tricalcium phosphate (beta-TCP) implanted in dog dorsal muscles. Specimens were harvested on days 14, 28, 42, 56, 112 and 168 after implantation, and were analyzed by hematoxylin and eosin (HE) staining, tartrate resistant acid phosphatase (TRAP) staining, immunohistochemistry, in situ hybridization, and silver impregnation. After day 28, abundant TRAP- and cathepsin K-positive multinucleated cells adhered to beta-TCP, suggesting that these cells are osteoclasts that can resorb beta-TCP. On day 56, new bone was formed and alpha1 chain of type I procollagen mRNA-positive osteoblasts lined the newly formed bone. Silver impregnation showed abundant collagen fibrils within the beta-TCP micropores. These results suggest that micropores function as a storage space for extracellular matrix components, including collagen. Newly formed bone never degenerated in the late stage, suggesting that beta-TCP has good biocompatibility and this material retains the conditions appropriate for osteointegration and bioresorption. In conclusion, beta-TCP has osteoinductivity after implantation in dog dorsal muscles without use of bone marrow cells or osteoinductive cytokines. The appearance of a large number of active osteoclasts precedes new bone formation. PMID- 16690122 TI - The effect of sterilisation on a poly(dimethylsiloxane)/poly(hexamethylene oxide) mixed macrodiol-based polyurethane elastomer. AB - The effect of various forms of sterilisation on a novel thermoplastic polyurethane elastomer synthesised using poly(hexamethylene oxide) (PHMO) and poly(dimethylsiloxane) (PDMS) macrodiols has been studied. The five sterilisation methods investigated were ethylene oxide (EtO) (single and multiple cycles), gas plasma, steam, vapour phase liquid chemical and gamma-irradiation (single and multiple cycles). Following sterilisation, scanning electron microscopy (SEM) and attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) were used to assess changes in the surface chemical structure and morphology, and gel permeation chromatography (GPC) and tensile testing were used to examine changes in bulk characteristics. Biostability was assessed using subcutaneous implantation of strained samples in sheep for 6 weeks. The results showed that the properties of the commercially available control material, Pellethane 2363 80A, were significantly affected by exposure to gamma-irradiation, steam and multiple cycles of EtO with aging and implantation compounding the effect. Exposure to a gas plasma sterilisation process resulted in significant degradation in both polyurethanes. A vapour phase liquid chemical sterilisation process caused minimal adverse effects. Sterilisation of the PDMS-based polyurethane using EtO, gamma-irradiation and autoclaving resulted in no significant changes in properties. The material's biostability was also unaffected by exposure to each of these sterilisation processes followed by short term implantation suggesting that this material is a potential candidate for use in a wide range of implantable medical devices sterilised using commercially available processes. Further biostability studies should be performed to assess the longer-term in vivo biostability of the PDMS-based material sterilised using autoclaving and gamma-irradiation. PMID- 16690123 TI - Quantitative two-photon Ca2+ imaging via fluorescence lifetime analysis. AB - Two-photon microscopy (TPM) revolutionized Ca2+ imaging by allowing recordings in the depth of intact tissue and live organisms. A serious limitation in TPM, however, is the lack of an accurate and straightforward approach for the quantification of Ca2+ signals, an ability that became an invaluable tool in fluorescence microscopy. Here, we present time-correlated fluorescence lifetime imaging (tcFLIM) as a ratiometric method for the quantification of Ca2+ signals in TPM. The fluorescence lifetime of the Ca2+-indicator dye Oregon Green BAPTA-1 (OGB-1) can be recorded using the approximately 80 MHz excitation pulses utilized in TPM. It shows a Ca2+ dependence that can be explained by the Ca2+-affinity, spectral properties and purity of the dye. Pixel-wise lifetime recordings, controlled by a laser-scanning microscope, allowed quantitative Ca2+ imaging in full-frame and linescan mode. Although we focused on the high-affinity Ca2+ indicator OGB-1, our tcFLIM-based quantification is applicable to other Ca2+ dyes and to fluorescence indicators in general. PMID- 16690124 TI - Oxidative stress in leukocytes from young prematurely aging mice is reversed by supplementation with biscuits rich in antioxidants. AB - Aging is associated with a progressive dysregulation of immune responses as a result of increased oxidative stress. Therefore, we have assessed the oxidative stress status of peritoneal leukocytes from young prematurely aging mice (PAM) as compared with non-prematurely aging mice (NPAM), as well as the effects on this oxidative stress of a dietary supplementation with biscuits rich in antioxidants (vitamin C, vitamin E, beta-carotenes, zinc and selenium). We found that, in the peritoneal leukocytes, the levels of several parameters of oxidation such as extracellular superoxide anion (O(2)(-)), Prostaglandin E(2) (PGE(2)), nitric oxide, oxidized glutathione (GSSG) and lipid peroxidation (malondialdehyde, MDA) were higher in PAM as compared with NPAM, whereas the antioxidant defences such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) activities, as well as reduced glutathione (GSH) levels, were decreased. Consequently, young PAM showed an oxidative stress in their leukocytes, which is characteristic of mice of an older chronological age. Antioxidant diet supplementation was able to restore redox homeostasis, increasing the antioxidant and decreasing the oxidant levels. Accordingly, supplementation with adequate levels of antioxidants, from an early age, could be useful to preserve health, especially in prematurely aging populations. PMID- 16690125 TI - Cloning and characterization of the dromedary (Camelus dromedarius) neonatal Fc receptor (drFcRn). AB - The full length cDNA of the dromedary neonatal Fc receptor (drFcRn) alpha chain was isolated and found that it is similar to the neonatal Fc receptor (FcRn) of other species with a high overall similarity to ruminant FcRn alpha chains. The drFcRn/Fc contact residues are highly conserved and predicted to bind both conventional (IgG1) and heavy chain (IgG2a, IgG3) antibodies. Using immunohistochemistry, we detected its expression in the hepatocytes and in epithelial cells of portal bile ductuli and also in the mammary gland acini and ducti. Remarkably, Ser313, that was identified to be crucial for apical to basolateral transcytosis, is substituted in the drFcRn alpha chain. The full length of the dog and orangutan FcRn alpha chains was also identified from databases. Analyzing the phylogenetic relatedness of this gene we found that dromedary clustered together with artiodactyls, dog is located between artiodactyls and primates, where the orangutan was branched, reflecting the accepted evolutionary relationships. PMID- 16690126 TI - Analysis of EST and lectin expressions in hemocytes of Manila clams (Ruditapes philippinarum) (Bivalvia: Mollusca) infected with Perkinsus olseni. AB - The hemocytes of invertebrates play key roles in both cellular and humoral immune reactions by phagocytosis or delivering immune factors such as lectin and anti microbial peptides. Bacterial infection causes changes in components such as lectins, anti-bacterial peptides, and lysosomal enzymes of plasma or hemolymph in molluscs. Previously, we found that infection with the protozoan parasite, Perkinsus, increases lectin synthesis in hemocytes. In order to investigate the patterns of genes expressed in Manila clams (Ruditapes philippinarum) infected with the protozoan parasite Perkinsus olseni, we constructed a cDNA library and sequenced 1850 clones (expressed sequence tags). A total of 79 ESTs, were related to 29 functional immune genes such as C-type lectin, lysozyme, and cystatin B, in Manila clams. Lectins were the largest group of immune-function ESTs found in our Manila clams library. Among 7 lectin clones, two full length cDNAs of lectins were cloned. MCL-3, which is a simple C-type lectin composed of 151 amino acids, has a relatively short signal sequence of 17aa and single carbohydrate recognition domain (CRD) of approximately 130 residues. It is highly homologous to eel C-type lectin. The sequence of mc-sialic acid-binding lectin consists of 168 amino acid residues with molecular weight of 19.2 and shows high homology to sialic acid-binding lectin from the snail, Cepaea hortensis. The expression of 7 different lectins in hemocytes was analyzed by RT-PCR using gene-specific primers. Hemocytes from Perkinsus-infected clam expressed different sets of lectins than with Vibrio infection. These results demonstrate that several lectins are involved in Manila clam innate immunity and different challenges induce expression of different lectins. PMID- 16690127 TI - Differential regulation of interleukin-1 receptor associated kinase 1 (IRAK1) splice variants. AB - IRAK family proteins play critical roles in regulating innate immunity. There are three differentially spliced variants of IRAK1, namely 1, 1b, and 1c. We and others have previously identified that the full length IRAK1 underwent covalent modifications such as phosphorylation and ubiquitination upon lipopolysaccharide challenge. In this report, we observed that IRAK1 could also undergo sumoylation which was responsible for its translocation into the nucleus. In contrast, IRAK1c remained stable and did not undergo modification upon various challenges. Furthermore, we showed that IRAK1c solely localized in the cytoplasm. IRAK1 was absent and IRAK1c was the primary form in human brains. The absence of full length IRAK1 and presence of IRAK1c may keep brain tissue in a resting non inflammatory state. Intriguingly, the full length IRAK1 form was consistently detected in brain tissues obtained from aged humans, suggesting that differential splicing of IRAK1 may correlate with the aging process. PMID- 16690128 TI - Fluocinolone acetonide implant (Retisert) for noninfectious posterior uveitis: thirty-four-week results of a multicenter randomized clinical study. AB - PURPOSE: To report the interim 34-week safety and efficacy results of a 3-year study to evaluate an investigational intravitreal fluocinolone acetonide (FA) implant in patients with noninfectious posterior uveitis. DESIGN: Prospective, dose-masked, dose-randomized, historically controlled, multicenter trial in patients with unilateral or bilateral disease. PARTICIPANTS: A total of 278 patients with recurrent noninfectious posterior uveitis were randomized to receive a 0.59-mg (n = 110) or 2.1-mg (n = 168) implant. In patients with bilateral disease, the more severely affected eye received the implant. METHODS: The implant was inserted surgically into the vitreous cavity through a pars plana incision. Follow-up visits were scheduled on day 2, week 1, and then every 4 to 6 weeks through 34 weeks after implantation. Systemic, periocular, and topical therapies were reduced as allowed by the clinical response. MAIN OUTCOME MEASURES: The primary efficacy outcome was a comparison of the recurrence rate in the implanted eye from the 34 weeks before implantation to the 34 weeks after implantation. Visual acuity (VA), need for adjunctive therapy, and safety also were assessed. RESULTS: Combining both doses, the FA implant reduced the rate of recurrences from 51.4% in the 34 weeks preceding implantation to 6.1% postimplantation (P<0.0001) in the study eyes. Comparatively, there was a significant increase in the recurrence rate in the fellow nonimplanted eyes from 20.3% preimplantation to 42.0% postimplantation (P<0.0001). Visual acuity was stabilized or improved in 87% of implanted eyes and generally was associated with reductions in the area of macular hyperfluorescence. The percentage of eyes that required systemic medications, periocular injections, and topical corticosteroids decreased from 52.9%, 63.0%, and 35.7%, respectively, preimplantation to 12.1%, 2.2%, and 16.5% postimplantation (P< or =0.0001 in all cases). At week 34, 51.1% of implanted eyes required ocular antihypertensive drops, and 5.8% underwent glaucoma filtering surgery. Lens opacity scores increased by > or =2 grades in 19.8% of phakic implanted eyes, and 9.9% required cataract surgery. There were no statistically significant differences in any of the parameters studied for the 0.59-mg implant, compared with the 2.1-mg implant. CONCLUSIONS: The FA implant significantly reduced uveitis recurrences, improved VA, and decreased the need for adjunctive therapy in the studied patient population. The most common side effects included increased intraocular pressure and cataract progression. PMID- 16690129 TI - Studies on the interaction of extended terpyridyl and triazine metal complexes with DNA. AB - The interaction with duplex DNA of a series of 12 previously synthesised ruthenium (II) complexes incorporating extended terpyridines or analogous ligands is reported. The interaction was investigated using absorption spectroscopy and also, when possible, luminescence spectroscopy. Binding parameters for the interactions were estimated using established models. It was found that binding affinities are relatively large and their magnitude is somewhat dependent on the metal centre attached to the ligand. Viscosity studies reveal that all the complexes interact with DNA through groove binding and not intercalation. The variation in binding affinities of these complexes is rationalised by a consideration of electrostatic and steric factors. PMID- 16690130 TI - Dioxygen affinities and catalytic oxidation activities of cobalt complexes with Schiff bases containing crown ether. AB - The stoichiometry of dioxygen uptake of Co complexes with 4',5'-bis-(5-chloro-2 hydroxyphenylmethylideneimino)benzo-12-crown-4 (1a), 4',5'-bis(2 hydroxyphenylmethylideneimino)benzo-12-crown-4 (1b), 4',5'-bis-(5-methoxy-2 hydroxyphenylmethylideneimino)benzo-12-crown-4 (1c), 4',5'-bis-(5-chloro-2 hydroxyphenylmethylideneimino)benzo-15-crown-5 (2a), 4',5'-bis(2 hydroxyphenylmethylideneimino)benzo-15-crown-5 (2b), 4',5'-bis(5-methoxy-2 hydroxyphenylmethylideneimino)benzo-15-crown-5 (2c) were measured in pyridine solution. The equilibrium constants and thermodynamic parameters (DeltaH0, DeltaS0) of these complexes were calculated. At the same time, their catalytic activities in cyclohexene oxidation to 2-cyclohexen-1-one were examined. The influence of the crown ether ring on the dioxygen affinity of Co complexes at different temperature and the catalytic activities were discussed in comparison with uncrowned analogous complexes. PMID- 16690131 TI - Streptococcus suis infection: a series of 41 cases from Chiang Mai University Hospital. AB - OBJECTIVES: The objectives of this study were (1) to assess the clinical manifestations, treatment, and outcome of Streptococcus suis infection in adult patients in northern Thailand, (2) to evaluate the anti-microbial sensitivity pattern and (3) to determine the predicting factors of high mortality rate. METHODS: A retrospective study was conducted at Chiang Mai University Hospital from May 2000 to December 2002. Anti-microbial susceptibility test was performed by agar disk diffusion and the minimal inhibitory concentration (MIC) by E-test. RESULTS: Forty-one patients (32 men and nine women, mean age 51 years) with S. suis infection were identified. Three patients had a history of exposure to pig or pork and one patient had a history of raw beef consumption. Clinical manifestations included infective endocarditis, meningitis, sepsis, spondylodiscitis, and endophthalmitis in 16, 13, 10, 1, and 1 patients, respectively. The overall mortality rate was 19.5%. On univariate analysis, low serum albumin, high serum total bilirubin, low platelet, and rapid onset of illness were significantly correlated with high mortality rate. All isolates were sensitive to penicillin (mean MIC90=0.027 microg/ml). CONCLUSIONS: S. suis infection is not uncommon in northern Thailand. High suspicion and early detection are important and could lead to the successful treatment. PMID- 16690132 TI - Immunovirologic consequences and safety of short, non-structured interruptions of successful antiretroviral treatment. AB - OBJECTIVES: The aim was to evaluate the safety of short antiretroviral treatment interruptions and their virologic and immunologic consequences in HIV-infected adults on highly active antiretroviral treatment (HAART) with suppressed viral replication. The viral efficacy upon reintroduction was also evaluated. PATIENTS AND METHODS: All patients with undetectable viral load while on HAART were prospectively followed to detect any treatment interruption. We analysed viral and cellular kinetics, incidence of resistance mutations, clinical outcome and results after therapy resumption. RESULTS: Twenty patients were included, mean time since HIV diagnosis was 95 months and time with undetectable viral load 16 months. Treatment was interrupted because of adverse effects, cancer, tuberculosis or patient will. Treatment was reintroduced after 4 weeks using, if possible, the same combination. HIV viral load was detectable on day 28 after interruption in 18 patients (90%). Median of CD4 cell count (p25-p75) decreased from 478/mm3 (96-716) to 257/mm3 (118-663) (p=0.5). Resistance mutations were found in 9 patients (45%) after interruptions. Treatment was reintroduced in 14 patients; all of them achieved viral suppression. CONCLUSIONS: In patients receiving HAART who have undetectable viral load, an interruption, no longer than 4 weeks, due to any intercurrent problem seems to be safe. Response to resumption can usually be achieved. Due to the frequent development of resistance, a genotypic test during interruption might be helpful. PMID- 16690133 TI - Cytomegalovirus: a cause of colonic stricture in a premature infant. AB - A 27-week-old infant developed symptoms of bowel obstruction when full enteral feeds were started. Laparotomy revealed strictures in the ascending and proximal transverse colon. Right hemicolectomy was performed. Histological examination of the resected large bowel demonstrated the presence of Cytomegalovirus inclusion bodies. Cytomegalovirus infections of the gut are extremely rare in neonates. This case report alerts neonatologists and microbiologists to consider Cytomegalovirus infection as a possible cause of bowel obstruction and necrotising enterocolitis like symptoms. PMID- 16690134 TI - Measuring dendritic distribution of membrane proteins. AB - Neurons perform much of their integrative work in the dendritic tree, and spinal motoneurons have the largest tree of any cell. Electrical excitability is strongly influenced by dendrite membrane properties, which are difficult to measure directly. We describe a method to measure the distribution of ion channel membrane densities along dendritic trajectories. The method combines standard immunohistochemistry with reconstruction procedures for both large-scale and small-scale optical microscopy. Software written for Matlab then extracts the colocalization of the target ion channel with the target dye injected cell, and calculates the relative channel density per square micron of cell surface area, as a function of distance from the cell body. The technique can be used to quantify the localization and distribution of any immunoreactive moiety, and the software provides a flexible vehicle for sensitivity analysis, to validate heuristics for selecting thresholds. PMID- 16690135 TI - Spatial embedding of neuronal trees modeled by diffusive growth. AB - The relative importance of the intrinsic and extrinsic factors determining the variety of geometric shapes exhibited by dendritic trees remains unclear. This question was addressed by developing a model of the growth of dendritic trees based on diffusion-limited aggregation process. The model reproduces diverse neuronal shapes (i.e., granule cells, Purkinje cells, the basal and apical dendrites of pyramidal cells, and the axonal trees of interneurons) by changing only the size of the growth area, the time span of pruning, and the spatial concentration of 'neurotrophic particles'. Moreover, the presented model shows how competition between neurons can affect the shape of the dendritic trees. The model reveals that the creation of complex (but reproducible) dendrite-like trees does not require precise guidance or an intrinsic plan of the dendrite geometry. Instead, basic environmental factors and the simple rules of diffusive growth adequately account for the spatial embedding of different types of dendrites observed in the cortex. An example demonstrating the broad applicability of the algorithm to model diverse types of tree structures is also presented. PMID- 16690136 TI - Low density and high affinity of platelet [3H]paroxetine binding in women with bulimia nervosa. AB - Impaired serotonin transmission has been suggested to be implicated in the pathophysiology of bulimia nervosa. As an indirect measure of brain serotonergic activity, the binding of tritiated ligands to platelet serotonin transporters has been studied in bulimia nervosa as well as in other putatively serotonin-related psychiatric disorders. In this study, the density and affinity of platelet serotonin transporters were assessed in 20 women meeting the DSM-IV criteria for bulimia nervosa and in 14 controls without previous or ongoing eating disorder using [(3)H]paroxetine as a ligand. In comparison to controls, women with bulimia nervosa had a significantly reduced number of platelet binding sites (B(max) = 721 +/- 313 vs. 1145 +/- 293 fmol/mg protein) and an increase in the affinity for the ligand demonstrated by a lower dissociaton constant (K(d) = 33 +/- 10 vs. 44 +/- 10 pM). A significant correlation between B(max) and K(d) values was found in patients but not in controls. Our results support the notion that bulimia nervosa is associated with a reduction in platelet serotonin transporter density. In addition, our study is the first to report that this reduced transporter density in women with bulimia nervosa is accompanied by an increase in the affinity of the transporter for the ligand. PMID- 16690137 TI - Use of a novel assay based on intact recombinant viruses expressing green (EGFP) or red (DsRed2) fluorescent proteins to examine the contribution of pol and env genes to overall HIV-1 replicative fitness. AB - Multiple studies have described a reduction in the replicative fitness of HIV-1 isolates harboring mutations that confer resistance to antiretroviral drugs. Contradictory results, however, have been obtained depending on the methodology used in each study (Quinones-Mateu, M.E., Arts, E.J., 2002. Fitness of drug resistant HIV-I: methodology and clinical implications. Drug Resist. Update 5, 224-233), affecting our understanding of the potential relationship of viral replicative fitness with HIV-1 disease. It has been demonstrated previously that both pol and env genes play a major role in HIV-1 replicative fitness of clinical isolates. Therefore, measuring clinically relevant replicative fitness using recombinant viruses where a single mutation and/or viral gene have been introduced does not seem like a reasonable approach in this era of multi-target antiretroviral therapy. A novel method was developed to measure HIV-1 replicative fitness based on recombinant viruses expressing the enhanced green fluorescent (EGFP) or the Discosoma sp. red fluorescent (DsRed2) proteins in a HIV-1NL4-3 backbone. Contrary to previous designs to analyze HIV-1 fitness, these replication competent viruses were created in an intact viral genetic background (without deleting or affecting the expression of any viral gene). This new system was used to evaluate the contribution of drug-resistance mutations in the pol and env genes to overall viral replicative fitness (in the presence and absence of drug pressure) using direct growth competition experiments. Mutations in pol showed a stronger effect on HIV-1 replicative fitness than mutations in the env gene associated with resistance to enfuvirtide, corroborating the plasticity of the later gene to accept mutations and the sensibility of the protease and reverse transcriptase enzymes to drug-associated primary mutations. In conclusion, a new protocol was used to measure HIV-1 replicative fitness in either the presence or absence of antiretroviral drugs, which may be used as a high-throughput assay to help us understand the clinical significance of viral fitness. PMID- 16690138 TI - Comparative analysis of molecular strategies attenuating positional effects in lentiviral vectors carrying multiple genes. AB - Efficient, high-level expression of multiple genes is often difficult to achieve in retroviral vectors, due to positional effects affecting transcription of adjacent sequences. Here we describe the comparative analysis of different strategies for co-expressing two model cDNA sequences in the context of a second generation lentiviral vector system. A first option was based on the generation of a polycistronic construct by subcloning an internal ribosome entry site (IRES) sequence between tandem cDNAs. IRES-dependent translation of the cDNA placed downstream (3') of the first transgene was poor, and the protein was barely detectable in transduced cells. A similar result was obtained when both transgenes were placed under the transcriptional control of two independent internal promoters. When these independent transcription units were separated by the 5'HS4 chromatin insulator of the chicken beta-globin locus, a marked increase of the expression of the downstream protein was observed. Similarly, insertion of a polyadenylation sequence between the tandem transcription units fully restored in transfection experiments - the expression of the downstream sequence, whose protein pattern was identical to the single-gene control, suggesting that in this specific construct transcriptional interference was the likely cause of the observed positional effects. These results indicate that chromatin insulator sequences can be useful molecular tools to overcome positional effects in the context of lentiviral vectors. PMID- 16690139 TI - A multiplex DNA suspension microarray for simultaneous detection and differentiation of classical swine fever virus and other pestiviruses. AB - An oligonucleotide suspension microarray (Luminex microsphere system) was developed for detection and differentiation of animal pestiviruses: classical swine fever virus (CSFV), bovine viral diarrhea virus types 1 and 2 (BVDV1 and BVDV2), and border disease virus (BDV). Species-specific and pestivirus-common oligonucleotide probes were designed to the 5' UTR region and conjugated to individual color-coded Luminex carboxy beads (probe beads). Target pestivirus sequences were amplified by asymmetric PCR using a biotinylated reverse primer and a forward and reverse primer ratio of 1:5. The biotinylated products were hybridized to eight probe beads in a multiplex assay and analyzed using streptavidin conjugated to a fluorescent reporter molecule. The assay was able to detect and differentiate all 40 strains of CSFV, BVDV1, BVDV2 and BDV tested. The analytical sensitivity was determined to be 0.2-10 TCID50/ml. The major advantages of the DNA-microsphere suspension microarray, as a low density array, are its ease of handling and ability to simultaneously detect and type multiple infectious agents. PMID- 16690140 TI - The interplay between antiviral activity, oligonucleotide hybridisation and nucleic acids incorporation studies. AB - Nucleoside analogues have been the most successful antiviral compounds. Likewise, they are the most intriguing antiviral compounds, because of their structural relationship to natural nucleosides. This is also the reason why the design process of a potential selective antiviral nucleoside is so difficult. Too many natural processes (from cellular uptake to DNA incorporation) and too many enzymes are involved in their biological effect (activity/toxicity/catabolism/anabolism) to make the design process readily predictable. The relationship between the physicochemical and biochemical properties of nucleoside analogues and their antiviral activity is very complex and could only be understood on a very long term basis. Here we try to explain some of the reasoning that was made during the design process leading to new potent antivirals with a phosphonate functionality. PMID- 16690141 TI - Identification of inhibitors to papillomavirus type 16 E6 protein based on three dimensional structures of interacting proteins. AB - Human papillomaviruses (HPV) cause cutaneous and genital warts. A subset of HPV types is associated with a high-risk for progression to malignancy. The E6 protein from the high-risk HPV types represents an attractive target for intervention because of its roles in viral propagation and cellular transformation. E6 functions in part by interaction with human cellular proteins, several of which possess a helical E6-binding motif. The role for each amino acid in this motif for binding E6 has been tested through structure determination and site-directed mutagenesis. These structural and molecular biological approaches defined the spatial geometry of functional groups necessary for binding to E6. This E6-binding information (the E6-binding pharmacophore) was transferred into a three-dimensional query format suitable for computational screening of large chemical databases. Compounds were identified and tested using in vitro and cell culture-based assays. Several compounds selectively inhibited E6 interaction with the E6-binding protein E6AP and interfered with the ability of E6 to promote p53 degradation. Such compounds provide leads for the development of new pharmacologic agents to treat papillomavirus infections and their associated cancers. PMID- 16690142 TI - Influence of light in acute toxicity bioassays of imidacloprid and zinc pyrithione to zooplankton crustaceans. AB - The acute toxicity of imidacloprid, a neonicotinoid insecticide, and zinc pyrithione (Zpt), a biocide used in anti-dandruff shampoos and protective antifouling paints, to three species of ostracods and two waterfleas, including Daphnia magna, was determined and compared under light and dark conditions. Under normal laboratory conditions, UV light had no significant influence on the outcome of toxicity bioassays, although in the case of imidacloprid both EC(50) and LC(50) calculated values were twice as high under the light as in the dark. No influence of UV light was observed on bioassays conducted with Zpt, in spite of the fast aqueous photolysis exhibited by this compound. Imidacloprid 48-h LC(50) for cladocerans (65-133mg/L) were two orders of magnitude higher than for ostracods (301-715microg/L); values of EC(50) for cladocerans and ostracods were 2-6mg/L and 3-16microg/L, respectively. Toxicity of Zpt to both ostracod and cladoceran species appears to be similar, with 48-h LC(50) in the range 137-524 and 75-197microg/L for ostracods and cladocerans, respectively, and similar values for EC(50)s. The mortality endpoint (LC(50)), however, is not a reliable predictor of the effects of imidacloprid under field situations (e.g. rice paddies), because the paralysis effect induced by this insecticide takes place at much lower concentrations than those required to cause the death of the animals: regardless of the taxa, differences as large as 100- or 600-fold were observed between the EC(50) and LC(50) for the same exposures. As a consequence, immobilization tests and EC(50) values are recommended for this class of compounds, while caution should be exercised in environmental risk assessments of this and possibly other related neonicotinoid insecticides with similar activity. PMID- 16690143 TI - Structure, function and evolution of the mitochondrial division apparatus. AB - Mitochondria are derived from free-living alpha-proteobacteria that were engulfed by eukaryotic host cells through the process of endosymbiosis, and therefore have their own DNA which is organized using basic proteins to form organelle nuclei (nucleoids). Mitochondria divide and are split amongst the daughter cells during cell proliferation. Their division can be separated into two main events: division of the mitochondrial nuclei and division of the matrix (the so-called mitochondrial division, or mitochondriokinesis). In this review, we first focus on the cytogenetical relationships between mitochondrial nuclear division and mitochondriokinesis. Mitochondriokinesis occurs after mitochondrial nuclear division, similar to bacterial cytokinesis. We then describe the fine structure and dynamics of the mitochondrial division ring (MD ring) as a basic morphological background for mitochondriokinesis. Electron microscopy studies first identified a small electron-dense MD ring in the cytoplasm at the constriction sites of dividing mitochondria in the slime mold Physarum polycephalum, and then two large MD rings (with outer cytoplasmic and inner matrix sides) in the red alga Cyanidioschyzon merolae. Now MD rings have been found in all eukaryotes. In the third section, we describe the relationships between the MD ring and the FtsZ ring descended from ancestral bacteria. Other than the GTPase, FtsZ, mitochondria have lost most of the proteins required for bacterial cytokinesis as a consequence of endosymbiosis. The FtsZ protein forms an electron transparent ring (FtsZ or Z ring) in the matrix inside the inner MD ring. For the fourth section, we describe the dynamic association between the outer MD ring with a ring composed of the eukaryote-specific GTPase dynamin. Recent studies have revealed that eukaryote-specific GTPase dynamins form an electron transparent ring between the outer membrane and the MD ring. Thus, mitochondriokinesis is thought to be controlled by a mitochondrial division (MD) apparatus including a dynamic trio, namely the FtsZ, MD and dynamin rings, which consist of a chimera of rings from bacteria and eukaryotes in primitive organisms. Since the genes for the MD ring and dynamin rings are not found in the prokaryotic genome, the host genomes may make these rings to actively control mitochondrial division. In the fifth part, we focus on the dynamic changes in the formation and disassembly of the FtsZ, MD and dynamin rings. FtsZ rings are digested during a later period of mitochondrial division and then finally the MD and dynamin ring apparatuses pinched off the daughter mitochondria, supporting the idea that the host genomes are responsible for the ultimate control of mitochondrial division. We discuss the evolution, from the original vesicle division (VD) apparatuses to VD apparatuses including classical dynamin rings and MD apparatuses. It is likely that the MD apparatuses involving the dynamic trio evolved into the plastid division (PD) apparatus in Bikonta, while in Opisthokonta, the MD apparatus was simplified during evolution and may have branched into the mitochondrial fusion apparatus. Finally, we describe the possibility of intact isolation of large MD/PD apparatuses, the identification of all their proteins and their related genes using C. merolae genome information and TOF-MS analyses. These results will assist in elucidating the universal mechanism and evolution of MD, PD and VD apparatuses. PMID- 16690144 TI - Multidisciplinary management in geriatric day-hospital is beneficial for elderly fallers: a prospective study of 28 cases. AB - We determined whether management including medical, psychological, and physiotherapeutic approaches, over a period of 6 weeks, has a beneficial effect on motor abilities, psychological status, and independence of elderly fallers with psychomotor disadaptation syndrome (PDS). We included 28 subjects (mean age 81.4 years). They were assessed from a medical, motor, and psychological point of view at both the inclusion and the end of the multidisciplinary intervention. A follow-up was conducted with multidisciplinary assessment at 6 and 9 months after the beginning of the study in order to evaluate duration of benefits of the management. The statistical analysis concerned only subjects who took part in the total multidisciplinary program, i.e., 14 subjects. The multidisciplinary intervention had an overall positive impact on motor abilities as shown by the increase in the mini-motor test scores, the rate of success in rising from the floor and decrease of time for the dual task. This study also showed a reduction in the fear of falling and a decrease in the rate of fallers. This positive effect on motor abilities, fear of falling and rate of fallers was sustained until 9 months after the beginning of the multidisciplinary management. This study shows the importance of a multidisciplinary management of elderly fallers with PDS. PMID- 16690145 TI - Edema of the paretic hand in elderly post-stroke nursing patients. AB - Post-stroke edema of the paretic hand constitutes an additional, functional, and esthetic nuisance for the patient. Although often encountered in daily practice, it is not even mentioned in the stroke chapters of the various textbooks. The phenomenon is far from being elucidated and various aspects are still obscure. In this study we tried to estimate the extent of post-stroke hand edema (PSHE) in a sample of elderly patients. The study group consisted of 188 elderly post-stroke nursing patients with hemi or only hand paresis. Seventy, age matched, non paretic patients were examined as controls. The basis of comparison was the difference in circumference between the two arms at three sites: mid-finger, hand, and wrist as measured in the control group. Values above two standard deviations (S.D.) of the mean difference in circumference of the controls, at two or three sites, were considered as edema. Edema of the paretic hand was detected in 37% of post-stroke patients. Most (about three-quarters), could be classified as simple PSHE, while the rest may have had reflex sympathetic dystrophy (RSD). PMID- 16690146 TI - The potential advantage of "no-touch" aortic technique in off-pump complete arterial revascularization. AB - OBJECTIVES: Off-pump coronary artery bypass grafting (OPCAB) and complete arterial revascularization without proximal anastomosis to the aorta may decrease neurological events after open-heart surgery. Few reports exist regarding the combination of OPCAB and complete arterial revascularization exploring the theoretical advantage of avoiding manipulation of the aorta. We review our results in 110 patients who underwent multiple grafts off-pump complete arterial revascularization. METHODS: 110 patients underwent multiple grafts OPCAB complete arterial revascularization, and were compared to 216 patients who underwent traditional multiple grafts on pump CABG. Preoperative renal failure was 12.7% (n=14) as compared to 5.1%, (n=11, p=0.01) in the control group and 33.6% (n=37) of the patients were 75 years or older as compared to 19.0% (n=41, p=0.003) in the control group. RESULTS: The mean number of grafts per patient undergoing multiple OPCAB complete arterial revascularization was 2.3, as compared to 3.11 in the control group (p<0.001). The mortality rate was 2.73% as compared to 1.85% (NS) in the control group. The incidence of CVA was 0% as compared to 2.31% (p=0.17) in the control group. CONCLUSIONS: Complete arterial OPCAB revascularization without manipulation of the aorta in high-risk patients can be performed with short-term similar results to conventional CABG and very low neurological complications. PMID- 16690147 TI - Are recessions good for workplace safety? AB - Empirical evidence shows that in recessions the rate of workplace accidents goes down. This paper presents a theory and an empirical investigation to explain this phenomenon. The theory is based on the idea that reporting an accident affects the reputation of a worker and raises the probability that he is fired. If the unemployment rate is high, a worker faces a big loss when fired and fewer workplace accidents are reported. The empirical investigation concerns workplace accidents in 16 OECD countries. We conclude that cyclical fluctuations in workplace accident rates have to do with reporting behavior of workers and not with changes in workplace safety. PMID- 16690148 TI - Relationship between ethanol tolerance, H+ -ATPase activity and the lipid composition of the plasma membrane in different wine yeast strains. AB - Ethanol tolerance, ATPase activity and the lipid composition of the plasma membrane to study potential relationship among them were examined in five different wine yeast strains. Yeast cells were subjected to ethanol stress (4% v/v). Principal component analysis of the results revealed that the wine yeasts studied can be distinguished in terms of ATPase activity and oleic acid (C18:1), and palmitoleic acid (C16:1), in plasma membrane. Multiple regression analysis was used to identify a potential influence of some components of the plasma membrane on ethanol tolerance and ATPase activity. Based on the results, the ergosterol, oleic acid and palmitoleic acid are highly correlated with ATPase activity and ethanol tolerance. Ethanol tolerance and the ATPase activity of the plasma membrane were correlated at the 96.64% level with the oleic acid and ergosterol in plasma membrane. The Saccharomyces cerevisiae var. capensis flor yeast strain, which exhibited the highest ergosterol concentration in plasma membrane when grown in the presence of 4% v/v ethanol, was found to be the most ethanol-tolerant. PMID- 16690149 TI - RFLP characterization of Aspergillus niger aggregate species from grapes from Europe and Israel. AB - In order to characterize by molecular methods the Aspergillus niger aggregate species involved in the ochratoxin A (OTA) contamination of European wine grapes and table grapes from Israel, a total of 173 strains were studied. The ITS-5.8S rDNA fragments of 173 A. niger agreggate strains from grapes included in this study were amplified and their PCR amplicons were RsaI digested in order to classify the strains in the RFLP types, N and T. All of the strains belonging to the A. niger aggregate were classified into the two RFLP types previously defined: type N (43%) and type T (57%). Twenty out of the 173 strains of A. niger aggregate produced OTA (0.1 to 10.5 mug g(-1)). All the OTA producing species belonged to the N-RFLP type. PMID- 16690150 TI - Surrogates for validation of electron beam irradiation of foods. AB - The aim of this study was to identify a potential surrogate to describe the radiation sensitivity of the most common pathogens encountered in fruits. Three pathogens: Escherichia coli O157:H7 933, Listeria monocytogenes ATCC 51414, and Salmonella Poona, and five non-pathogens: E. coli K-12 MG1655, Listeria innocua Seeliger 1983 (NRRL B-33003 and NRRl B-33014), Enterobacter aerogenes, and Salmonella LT2 were inoculated (populations of 10(7)-10(9) CFU/ml) into model food systems (10% w/w gelatin) and exposed to doses up to 1.0 kGy using a 2 MeV Van der Graaf linear accelerator. The non-pathogen E. coli K-12 MG1655 was highly resistant to radiation (D(10)=0.88 kGy) in comparison to the other strains while L. monocytogenes was the more radiation-resistant pathogen (D(10)=1.09 kGy). Thus, E. coli K-12 MG1655 could be a suitable surrogate for e-beam studies with L. monocytogenes as the indicator pathogen. L. innocua strains were more radiation-sensitive (D(10)=0.66, 0.72 kGy) than their pathogenic counterpart. S. Poona and E. coli O157:H7 were even more radiation-sensitive (D(10)=0.38, 0.36 kGy, respectively). S. LT2 was the least radiation-resistant pathogen with D(10)=0.12 kGy. In a later study, the radiation resistance of the pathogens and the surrogate was evaluated when inoculated in a real food (i.e., fresh cantaloupe). The D(10) values obtained in this experiment were higher than those obtained with the model foods. However, the surrogate was still more radiation resistant and could therefore be used to indicate decontamination of the target pathogens under electron beam irradiation. PMID- 16690151 TI - Salmonella detection in probiotic products. AB - The presence of large amounts of probiotic bacteria in a sample may interfere with the detection of undesirable microorganisms. To illustrate this, infant formula with various strains of probiotic bacteria and the corresponding probiotic culture powders and premixes were artificially contaminated with low levels of Salmonella. Recovery of Salmonella was generally very poor when the conventional pre-enrichment procedure using buffered peptone water (BPW) was applied. However, this problem was overcome by adding antimicrobial compounds to selectively suppress the growth and/or metabolic activity of the probiotic bacteria and increasing the buffering capacity of the pre-enrichment broth. It is recommended that these analytical constraints are already addressed during the development phase of new probiotic products. PMID- 16690152 TI - Genetic relatedness between Listeria monocytogenes isolates from seafood and humans using PFGE and REP-PCR. AB - Listeria monocytogenes has been isolated from catfish and various non-catfish seafoods. Despite progress that was made to understand the relationship between L. monocytogenes isolated from seafood and humans, no study has emphasized the genetic relatedness between catfish and non-catfish seafood and human isolates. The objectives of this study were to (1) investigate the genetic relationship between L. monocytogenes isolates from catfish, non-catfish seafood and humans using pulsed-field gel electrophoresis (PFGE) and repetitive extragenic palindromic element-based PCR (REP-PCR) linking with serotypes; and (2) compare the distinct capabilities of these methods. The genetic relationship among 36 catfish, 35 non-catfish seafood, and 57 human isolates were analyzed using Bionumerics software. Among a total of 128 isolates, only two subtypes by PFGE with ApaI-digestion, one subtype by PFGE with AscI-digestion and three subtypes by REP-PCR were shared by non-catfish seafood or catfish with human isolates. Although most of the non-catfish seafood and catfish isolates were genetically distinct from human isolates, the human isolates used in this study may not be truly representative of all clinical isolates. Serotype 4b was dominant in human isolates, whereas serotypes 3b, 1/2a, and 1/2b and serotypes 4b, 1/2a and 1/2b were commonly found in catfish and non-catfish seafood, respectively. Furthermore, 97, 87 and 94 subtypes of L. monocytogenes were revealed using the ApaI-digestion, the AscI-digestion, and the REP-PCR, respectively. Their respective discriminatory indexes were 0.994, 0.990 and 0.994. Distinct genetic groups based on the difference of flagella antigens were observed in all three methods. The study suggests that the REP-PCR possesses a similar discriminatory ability as the PFGE for subtyping L. monocytogenes. Therefore, the REP-PCR that is rapid and less expensive could be considered as an alternative method for subtyping L. monocytogenes. PMID- 16690153 TI - Modeling the frequency and duration of microbial contamination events. AB - The frequency and duration of microbial contamination events in the environment in which ready-to-eat (RTE) foods are exposed for processing and packaging is subject to uncertainty and variability. Variability, within-model parameter uncertainty, and uncertainty regarding model selection are formally considered in modeling the frequency and duration of such contamination events by Listeria species. The estimated duration of contamination events represents a case where variability dominates with relatively little uncertainty about parameter values or model form. The estimated frequency of contamination events represents a case where there is not only substantial variability but also considerable within model parameter uncertainty, as well as some uncertainty regarding model selection. The Bayesian Information Criterion provides a formal way of taking into account model uncertainty. PMID- 16690154 TI - Efficiency of electrolyzed oxidizing water on reducing Listeria monocytogenes contamination on seafood processing gloves. AB - Food processing gloves are typically used to prevent cross-contamination during food preparation. However, gloves can be contaminated with microorganisms and become a source of contamination. This study investigated the survival of Listeria monocytogenes on gloves and determined the efficacy of electrolyzed oxidizing (EO) water for reducing L. monocytogenes contamination on seafood processing gloves. Three types of reusable gloves (natural rubber latex, natural latex, and nitrile) and two types of disposable gloves (latex and nitrile) were cut into small pieces (4 x 4 cm(2)) and inoculated with 5-strain L. monocytogenes cocktail (5.1 x 10(7) CFU/cm(2)) with and without shrimp meat residue attached to surfaces. L. monocytogenes did not survive well on clean reusable gloves and its populations decreased rapidly to non-detectable levels within 30 min at room temperature. However, high levels of Listeria cells were recovered from clean disposable gloves after 30 min of inoculation. Presence of shrimp meat residue on gloves enhanced the survival of L. monocytogenes. Cells of L. monocytogenes were detected on both reusable and disposal gloves even after 2 h at room temperature. Soaking inoculated gloves in EO water at room temperature for 5 min completely eliminated L. monocytogenes on clean gloves (>4.46 log CFU/cm(2) reductions) and significantly (p<0.05) reduced the contamination on soil-containing gloves when compared with tap water treatment. EO water could be used as a sanitizer to reduce L. monocytogenes contamination on gloves and reduce the possibility of transferring L. monocytogenes from gloves to RTE seafoods. PMID- 16690155 TI - The development of 'feeder' cells for the preparation of clinical grade hES cell lines: challenges and solutions. AB - The development of human embryonic stem cell (hESC) lines for research and therapy is hampered by the need to improve the basic methodologies for cell culture expansion. In most current methods hESC lines are cultured on a mouse or human feeder cell layer which appears to be the most reliable way to maintain cells stably in the undifferentiated state. However, co-culture introduces complications for studying stem cell biology and the delivery of safe therapies for the future. This article reviews the specific risks associated with any proposed clinical use of feeder cells of mouse origin and compares these with the benefits and risks of using human feeder cells. The further work required to establish clinical grade feeder cell lines for hESC line culture is significant and costly. Much work is being done to find feeder-free culture systems but these are at an early stage of development and there may be consequences that affect the value of the hESCs for research and development. These challenges should be viewed in the context of the huge amount of work that will be required over many years to develop robust differentiation protocols and establish fully defined procedures and adequate safety data for embryonic stem cell products. PMID- 16690156 TI - Impact of integrated positron emission tomography and computed tomography on staging and management of gallbladder cancer and cholangiocarcinoma. AB - BACKGROUND/AIMS: (1) To evaluate the diagnostic value of integrated positron emission and computed tomography (PET/CT) in comparison with contrast-enhanced CT (ceCT) to detect biliary tract tumors and associated distant and regional lymph node metastases and (2) to evaluate the impact of PET/CT on therapy management. METHODS: From January 2001 to March 2005, each patient who was treated for a malignancy of the biliary tract underwent PET/CT examination in addition to the standard work-up imaging. Data were prospectively collected and analyzed in comparison with ceCT. RESULTS: Sixty-one patients with malignancies of the biliary tract were included into the study. Diagnosis was proven in all patients either by histology or cytology. PET/CT detected all gallbladder cancers (n=14). PET/CT and ceCT provided a comparable accuracy for the primary intra- (n=14) and extra-hepatic cholangiocarcinomas (n=33). All distant metastases (12/12) were detected by PET/CT, but only 3/12 by ceCT (p<0.001). Regional lymph node metastases were detected by PET/CT and ceCT in only 12% vs. 24%. PET/CT findings resulted in a change of management in 17% of patients deemed resectable after standard work-up. CONCLUSIONS: PET/CT is particularly valuable in detecting unsuspected distant metastases which are not diagnosed by standard imaging. Thus, PET/CT staging has an important impact on selection of adequate therapy. PMID- 16690157 TI - Conflicting candidates for cattle QTLs. AB - Genome scans have identified quantitative trait loci (QTLs) affecting milk yield and composition in dairy cattle. For one QTL on bovine chromosome 6 (BTA6), previously fine-mapped to a 420-Kb region, mutations in two different genes (OPN and ABCG2) have been proposed as the underlying functional mutation. Comparing the arguments for each gene suggests that both mutations are equally probable. However, functional studies and/or additional populations are required to provide a definite answer. PMID- 16690158 TI - Oxymoron no more: the expanding world of heterochromatic genes. AB - Heterochromatin has been oversimplified and even misunderstood. In particular, the existence of heterochromatic genes is often overlooked. Diverse types of genes reside within regions classified as constitutive heterochromatin and activating influences of heterochromatin on gene expression in Drosophila are well documented. These properties are usually considered paradoxical because heterochromatin is commonly portrayed as "silent chromatin". In the past, studies of heterochromatic genes were limited to a few Drosophila genes. However, the recent discovery of several hundred heterochromatic genes in Drosophila, plants and mammals through sequencing projects offers new opportunities to examine the variety of ways in which heterochromatin influences gene expression. Comparative genomics is revealing diverse origins of heterochromatic genes and remarkable evolutionary fluidity between heterochromatic and euchromatic domains. These features justify a broader view of heterochromatin, one that accommodates repressive, permissive and activating effects on gene expression, and recognizes chromosomal and evolutionary transitional states between heterochromatin and euchromatin. PMID- 16690159 TI - Changes in the distribution of lung cancer cell types and patient demography in a developing multiracial Asian country: experience of a university teaching hospital. AB - A comparison of patients with lung cancer diagnosed at the University of Malaya Medical Centre, Kuala Lumpur, Malaysia from October 1991 to September 1999 with another group of lung cancer patients diagnosed at the same hospital during an earlier period of 1967-1976 was undertaken to determine whether there had been a change in the distribution of lung cancer cell types and patient demography. The number of histologically and/or cytologically proven lung cancer cases was 583 from October 1991 to September 1999 and 278 from 1967 to 1976. The mean (S.D.) age of the patients during the period 1991-1999, 60.1 (12.0) years was similar to that of patients during the period 1967-1976, 60.3 (12.2) years. There was no shift of the peak age distribution of lung cancer (i.e., the 7th decade) between the two periods. In the recent period, the percentage of patients with adenocarcinoma had increased significantly to 43.2% from 25.2% while that of large cell carcinoma had decreased to 3.3% from 11.9%. The percentages of patients with squamous cell carcinoma (SCC) and small cell lung cancer remained stable. In the period 1967-1976, SCC was the commonest cell type in men and in smokers while adenocarcinoma was the commonest cell type in women and in never smokers. In the period 1991-1999, adenocarcinoma was the commonest cell type in both men and women as well as in smokers and never smokers. PMID- 16690160 TI - Would alternate tobacco products use be better than smoking? AB - Tobacco use is dominated by cigarette smoking. Most toxicity due to cigarette smoking is related to the burning process. Several observations suggest that unburned or oral tobacco is less harmful than cigarette smoking. Up to now, physicians have advised a policy based on two ideas: (i) young people should not start smoking and (ii) the only choice for smokers is to quit. However, numerous smokers cannot or do not want to give up; eventually, one out of two lifelong smokers will die from smoke-related diseases. Thus, it is legitimate to ask if alternate nicotine containing product use would be better than cigarette smoking and limit the tobacco death toll. However, the logic leading to the concept of tobacco harm reduction (THR) turns out to be much more complex than it looks at a first glance. PMID- 16690161 TI - Solitary pulmonary nodules: association between signal characteristics in dynamic contrast enhanced MRI and tumor angiogenesis. AB - PURPOSE: To estimate the association between signal characteristic of dynamic enhanced MRI using curve types and angiogenesis in solitary pulmonary nodules. MATERIALS AND METHODS: Thirty-six patients with a solitary pulmonary nodule (SPN) ranging in size from 6 to 37 mm (median 17 mm) underwent dynamic contrast enhanced MRI with a time interval of 10 s over a total period of 4 min. Resulting from the time-intensity curves four different enhancement curve profiles (A-D) were defined: type A with strong increase followed by early washout, type B with strong increase without washout, type C with slow increase and type D without relevant increase. Maximum peak (Pmax), slope of the first bolus transit (slope) and washout were calculated. Microvessel densities (MVD) were counted at the margins and at the center of the nodules. The mean MVD of each nodule was calculated. Enhancement characteristics were correlated with MVD grouped by diagnosis and by curve types. Curve types were correlated with the score of vascular endothelial growth factor (VEGF). RESULTS: The frequency of malignancy was 55% (20/36). Using curve types for differentiation between malignant and benign SPN, the sensitivity, specificity and accuracy were 100%, 75% and 89%, respectively. The correlation between Pmax and MVD(mean) for all nodules was moderate (r(s)=0.4, P=0.02). A relevant correlation was found between Pmax and MVD(margin) in curve type A (r(s)=0.63; P=0.04) and Pmax and MVD(mean) in curve type C (r(s)=0.86; P=0.006). No positive correlation was found between Pmax and MVD (mean, center and margin) in curve type B. No significant correlation was found for slope and washout. VEGF score correlated positively with curve types (r(s)=0.67; P<0.001). CONCLUSION: A relevant association between perfusion curve profiles and angiogenesis was found in malignant nodules having early washout and in benign lesion with a slow increase of enhancement. In cases of strong signal increase without washout additional factors for enhancement must be considered. The use of curve profiles could allow for the estimation of the extent of VEGF. PMID- 16690162 TI - Retardation of organo-bromides in a fractured chalk aquitard. AB - This study investigates the mechanisms controlling the distribution of 3-bromo 2,2-bis(bromomethyl)propanol (TBNPA) and 2,2-bis(bromomethyl)propan-1,3-diol (DBNPG) in a fractured chalk aquitard. An extensive monitoring program showed a systematic decrease in the TBNPA/DBNPG ratio with distance from the contamination source. Sorption of TBNPA on the white and/or gray chalks comprising the aquitard is approximately one order of magnitude greater than that of DBNPG. This results in more efficient removal of TBNPA from the fracture into the porous matrix and thus decreases the TBNPA/DBNPG ratio in the fracture water. Mathematical modeling of solute transport in the fracture domain illustrates the probable importance of sorption in controlling the spatial variation in TBNPA and DBNPG ratio. PMID- 16690163 TI - Salicylic acid induced changes on some physiological parameters symptomatic for oxidative stress and mineral nutrition in maize (Zea mays L.) grown under salinity. AB - It has been proposed that salicylic acid (SA) acts as an endogenous signal molecule responsible for inducing abiotic stress tolerance in plants. The effect of varying salicylic acid (SA) supply (0, 0.1, 0.5 and 1.0mM) on growth, mineral uptake, membrane permeability, lipid peroxidation, H(2)O(2) concentration, UV absorbing substances, chlorophyll and carotenoid concentrations of NaCl (40 mM) stressed maize (Zea mays L.) was investigated. Exogenously applied SA increased plant growth significantly both in saline and non-saline conditions. As a consequence of salinity stress, lipid peroxidation, measured in terms of malondialdehyde (MDA) content and membrane permeability was decreased by SA. UV absorbing substances (UVAS) and H(2)O(2) concentration were increased by increasing levels of SA. SA also strongly inhibited Na(+) and Cl(-) accumulation, but stimulated N, Mg, Fe, Mn and Cu concentrations of salt stressed maize plants. These results suggest that SA could be used as a potential growth regulator to improve plant salinity stress resistance. PMID- 16690164 TI - Differential expression of wheat aspartic proteinases, WAP1 and WAP2, in germinating and maturing seeds. AB - Two aspartic proteinase (AP) cDNA clones, WAP1 and WAP2, were obtained from wheat seeds. Proteins encoded by these clones shared 61% amino acid sequence identity. RNA blotting analysis showed that WAP1 and WAP2 were expressed in both germinating and maturing seeds. The level of WAP2 mRNA expression was clearly weaker than that of WAP1 in all tissues of seeds during germination and maturation. APs purified from germinating seeds were enzymatically active and digested the wheat storage protein, gluten. To elucidate the physiological functions of WAP1 and WAP2 in seeds, we investigated the localisation of WAP1 and WAP2 by in situ hybridisation. In germinating seeds investigated 24h after imbibition, both WAP1 and WAP2 were expressed in embryos, especially in radicles and shoots, scutellum, and the aleurone layer. In maturing seeds, WAP1 was expressed in the whole embryo, with slightly stronger expression in radicles and shoots. WAP1 was also expressed in the aleurone layer 3 weeks after flowering. Strong signals of WAP1 mRNA were detected in the whole embryo and aleurone layer 6 weeks after flowering. On the other hand, WAP2 was scarcely detected in seeds 3 weeks after flowering, and thereafter weak signals began to appear in the whole embryo. WAP1 and WAP2 were expressed widely in germinating and maturing seeds. Such diversity in site- and stage-specific expression of the two enzymes suggests their differential functions in wheat seeds. PMID- 16690165 TI - Allantoin has a limited role as nitrogen source in cultured coffee cells. AB - In plants the ureides allantoin (ALN) and allantoic acid (ALA) are formed in purine metabolism, and in some legumes both compounds play an important role as nitrogen (N) sources. In coffee plants, ALN and ALA are catabolites of caffeine degradation. Caffeine is found throughout the coffee plant and in some parts this alkaloid can accumulate up to 4% dry basis. Therefore, caffeine degradation via ureides may make an important contribution to N metabolism of the plant. Using coffee cell suspension as a model we investigated the contribution of ALN as a source of N in coffee. ALN was incorporated in the liquid medium and after 20 d of cultivation, cell mass, NO(3), NH(4), amino acids, soluble proteins, ALN and caffeine were determined in the cells. The activity of glutamine synthetase was also studied. The results showed that despite being taken up by cells ALN does not contribute significantly as a source of N in coffee cells. Compared with mineral N sources, cells grown with ALN-N accumulated much less mass. The inclusion of ALN in the medium caused significant alterations in the content of some N compounds indicating a stress condition. PMID- 16690166 TI - Physiology and biochemistry of source-regulated protein accumulation in the wheat grain. AB - Wheat is unique among cereals for the baking qualities of its flour, which are dependent upon the type and concentration of its proteins. As a consequence, the grain protein concentration (GPC) is one of the main determinants of wheat international market price. More than 50-70% of the final grain N is accumulated before flowering and later remobilized to the grain, N fertilization being the common practice used to produce high GPC. However, after incremental additions of N fertilizer, GPC reaches a maximum and then remains constant, without any increase in N uptake or remobilization by the crop, thus decreasing the efficiency of N fertilizer. Although, the genetic and molecular mechanisms that regulate N uptake by the roots are being clarified quickly, the regulation and physiology of N transport from the leaves to the grain remains less clear. In this review, the possible regulatory points involved in N transport to the grain and the difficulties for increasing GPC are discussed. It has been demonstrated that protein synthesis in the grain is source-limited, and that the grain can accumulate protein limited only by the amino acids provided by the phloem. It has also been shown that there is no limitation in the amino acid/sugar ratios that can be exported to the phloem. On the other hand, NO(3)(-) uptake transporters are depressed when the plant concentration of some amino acids, such as glutamine, is high. It has also been shown that a high N supply increases cytokinins concentration, preventing leaf senescence and proteolysis. Based on this information, it is postulated that there are two main regulatory points during grain filling when plant N status is ample. On the one hand, the N uptake transporters in the roots are depressed due to the high amino acids concentration in the tissues, and N uptake is low. On the other, a high amino acids concentration keeps the cytokinins level high, repressing leaf protein degradation and decreasing amino acid export to the phloem. As a consequence, GPC cannot be increased despite the ample N supply. PMID- 16690167 TI - Protective effect of nitric oxide on iron deficiency-induced oxidative stress in maize (Zea mays). AB - The effects of nitric oxide (NO) in protecting maize (Zea mays) leaves against iron deficiency-induced oxidative stress were investigated. The increased contents of hydrogen peroxide (H(2)O(2)) and superoxide (O(2)(-)*) due to iron deficiency suggested oxidative stress. The increased contents of thiobarbituric acid-reacting substances (TBARS) and the decreased contents of protein-bound thiol (PT) and non-protein-bound thiol (NPT) indicated iron deficiency-induced oxidative damage on proteins and lipids. Sodium nitroprusside (SNP), a nitric oxide (NO) donor, partially reversed iron deficiency-induced retardation of plant growth as well as chlorosis. Reduced contents of H(2)O(2), O(2)(-)*, TBARS and increased contents of PT and NPT also indicated that NO alleviated iron deficiency-induced oxidative damage. The activities of SOD and GR decreased sharply while the activities of CAT, POD and APX increased under SNP treatment. Our data suggest that NO can protect maize plants from iron deficiency-induced oxidative stress by reacting with ROS directly or by changing activities of ROS scavenging enzymes. PMID- 16690168 TI - Enhanced sensitivity to oxidative stress in an Arabidopsis nitric oxide synthase mutant. AB - The possible involvement of nitric oxide (NO) in oxidative stress tolerance was studied using Arabidopsis thaliana wild type (WT) and Atnos1 mutant plants, in which endogenous NO production is greatly diminished because 80% of nitric oxide synthase (NOS) activity is eliminated due to T-DNA insertion in the first exon of the NOS1 gene. Compared with WT, Atnos1 mutant plants showed increased hypersensitivity to salt stress and methyl viologen (MV) treatment. The maximal photochemical efficiency of photosystem II (F(v)/F(m)) and membrane integrity decreased in WT and Atnos1 mutant plants under stresses, but the extent was higher in the mutant. Treatment with sodium nitroprusside (SNP) (a NO donor) to Atnos1 mutant plants alleviated the damage. Instead, inhibition of nitric oxide accumulation in the WT plants produced opposite effects. Hydrogen peroxide and lipid peroxidation increased and the extent was higher in Atnos1 mutant plants than that in WT plants under MV stress. These results indicated that nitric oxide could protect the damage against NaCl and MV treatments. PMID- 16690169 TI - Roots and leaves display contrasting oxidative response during salt stress and recovery in cowpea. AB - In this study, we compare some antioxidative responses of leaves and roots associated to growth reduction in cowpea plants (Vigna unguiculata) during short term salt stress and recovery. The salt treatment was imposed (200 mM NaCl) for six consecutive days and the salt withdrawal after 3 d. The salt treatment caused an almost complete cessation in the relative growth rate of both leaves and roots. Although NaCl withdrawal has induced an intense reduction in the Na(+) content from the leaves and roots, the growth recovery was slight, after 3 d. The leaf lipid peroxidation was increased in salt-stressed plants and slightly reduced in recovered plants after 3 d. Surprisingly, in the salt-stressed roots it decreased markedly after 3 d treatment and in the pre-stressed/recovered roots it was restored to levels near to the control. In leaves, catalase (CAT) activity showed a rapid and prominent decrease after 1 d of NaCl treatment and salt withdrawal had no effect on its recovery. In contrast, the root CAT activity was not changed by effects of both NaCl and salt withdrawal, over time interval. Leaf superoxide dismutase (SOD) activity did not change in all treatments, whereas in roots it significantly decreased after 3 d of salt treatment and recovered after NaCl withdrawal. Contrasting to the other enzymes, the guaiacol-peroxidase activity increased in leaves and roots, reaching almost 200% of control values and it significantly decreased in both organs from the pre-stressed/recovered plants. In conclusion, cowpea roots and leaves present distinct mechanisms of response to lipid peroxidation and CAT and SOD activities during salt stress and recovery. However, these responses and/or the oxidative damages caused by reactive oxygen species were not related with the growth reduction. PMID- 16690170 TI - A phosphonamidate containing aromatic N-terminal amino group as inhibitor of leucine aminopeptidase-design, synthesis and stability. AB - Fully deprotected phosphonamidate dipeptides, predicted as effective inhibitors of cytosolic leucine aminopeptidase, showed unexpected instability in water solution at pH below 12. Their hydrolysis rate was strictly correlated with basicity of the N-terminal amino group. To improve this feature a phosphonamidate analogue containing less basic, aromatic 2-aminophenylphosphonate residue in P1 position of the inhibitor was designed. The target compound was synthesised starting from diethyl 2-nitrophosphonate in several step procedure. The decrease in basicity of the terminal amino moiety of the modified analogue in fact resulted in satisfactory improvement of hydrolytic stability of the P-N bond. The developed phosphonamidate was proved to be fully resistant to hydrolysis above pH 7. Surprisingly, tested in enzymatic assays towards leucine aminopeptidase (optimum pH 8.5), it did not exhibit inhibition activity up to milimolar concentration. The explanation could be that diminishing the basic character of the terminal amino group may result in a change of its affinity towards the zinc ions. PMID- 16690171 TI - [Hepatitis C-autoimmune hepatitis overlap syndrome: five new cases]. AB - PURPOSE: The hepatitis C-Autoimmune hepatitis overlap syndrome is an uncommon condition whose management can be difficult in both diagnosis and treatment. PATIENTS AND METHODS: Five cases of hepatitis C and autoimmune hepatitis overlap syndrome, brought by a retrospective study, are reported. Hepatitis C was proven by a positive HCV viral load and the autoimmune hepatitis was proven by characteristic immunological and/or histological features. RESULTS: All patients were female, the mean age at diagnosis was 54.2 years (+/-6.6), only one patient had a history of autoimmune disease (autoimmune thyroiditis). Four patients had significant autoantibodies levels (over 1/320) and 4 had a high serum gammaglobulin level (over 1.5 times the upper normal limit). Liver biopsy showed characteristic features of autoimmune hepatitis in all cases and characteristic features of chronic HCV infection in 3 cases. Corticosteroid and/or immunosuppressive treatment was given as a first line therapy in 4 cases while an antiviral treatment has been tried in 4 cases with a good viral response in 2 of them. Corticosteroid and/or immunosuppressive treatment enhanced HCV viral load in all cases, however a histological improvement was observed in every case in which a control biopsy has been performed (3 cases). CONCLUSION: This study highlights the deciding contribution of the initial histological findings in the diagnosis of such a HCV/autoimmune hepatitis overlap syndrome; it also demonstrates that histological outcome is not necessarily compromised by corticosteroid and/or immunosuppressive treatment. PMID- 16690173 TI - [Iatrogenic angioedema: clinical features]. PMID- 16690174 TI - REMOVED: l-Carnitine and alpha-lipoic acid improves mitochondrial function during ageing process. AB - This article has been removed consistent with Elsevier Policy on Article Withdrawal. The Publisher apologies for any inconvenience this may cause. PMID- 16690175 TI - Total serum homocysteine, folate and vitamin B12 in a Greek school age population. AB - BACKGROUND & AIMS: Moderate hyperhomocysteinemia is an independent risk factor for cardiovascular disease (CVD) even among children. The purpose of this study is to investigate for the first time the distribution and determinants of total serum homocysteine (tHcy) levels in healthy Greek children. METHODS: tHcy, folate, B12 were measured in 524 children (275 boys and 249 girls) aged 6-15 years old from different socioeconomic status in Northern Greece. RESULTS: The geometric mean tHcy level for boys and girls was 7.8 (3.4-24.2) and 7.5 (3.9 29.0) micromol/L, respectively. Eighty one (15.4%) children had homocysteine levels above the upper reference limits (>10 micromol/L). The geometric mean serum tHcy level was significantly (P<0.001) increasing with age; 6.4 (3.4-11.2) micromol/L was found in the age group of 6-9 yr (group1), 7.2(4.1-22.1) micromol/L in the one of 10-12 yr (group 2) and 8.5 (3.9-29.0) micromol/L in the one of 13-15 yr (group 3). Serum folate levels were found to be statistically significant (P<0.001) between age group 1 and age group 3 [11.8 (4.66-20.00) vs. 7.5 (0.99-20.00)ng/mL) and between age group 2 and 3 [10.0 (1.82-20.0) vs. 7.5 (0.99-20.00)ng/mL]. Vitamin B12 levels were significantly (P<0.001) different in the three age groups [1048 (117-2000), 805 (296-2000), 700 (214-2000)pg/mL] respectively. Age, BMI, waist circumference (WC), systolic blood pressure (SBP) and diastolic blood pressure (DBP) were positively correlated with tHcy, whereas serum folate and vitamin B12 were negatively correlated. No association was found between tHcy levels and parental education status. In multiple linear regression analysis only age (Beta: 0.248, 95%, CI: (0.159-0.361), P<0.05) and folate (Beta: 0.347, 95%, CI: [(-0.206)-(-0.118)], P<0.05) were found significantly and independently associated with tHcy. CONCLUSIONS: tHcy levels were increasing with age and boys were found to have slightly higher levels than girls. Age and folate levels were the most significantly and independently determinants associated with tHcy. Children with tHcy levels above the upper reference limits (>10 micromol/L) were found to be correlated with BMI, WC, SBP, serum folate and vitamin B12 levels. These children should be encouraged to include high folate food items in their diet and where necessary folate supplements should be recommended. In addition, more prospective studies are necessary in order to evaluate the relationship of tHcy and CVD risk factors in children of our region. PMID- 16690176 TI - Magnesium and diabetes mellitus: their relation. AB - The aim of this review was to elaborate a synthesis about the discussions on magnesium and diabetes mellitus, in the last 14 years. The magnesium deficiency has been associated with chronic diseases, amongst them, diabetes mellitus. Epidemiological studies had shown low levels of magnesium ingestion in the general population, as well as a relation between the ingestion of food rich in magnesium and the reduction of diabetes installation and its complications. Hypomagnesemia is frequently present in diabetic patients, however there is not an exact elucidation of the mechanism of magnesium deficiency in diabetes mellitus. On the other hand, in the presence of this illness, it is observed that inadequate metabolic control can affect the corporal concentrations of magnesium, developing hypomagnesemia, which may be still directly related with some micro and macrovascular complications observed in diabetes, as cardiovascular disease, retinopathy and neuropathy. This way, the chronic complications of diabetes can appear precociously. Based on this, the supplementation with magnesium has been suggested in patients with diabetes mellitus who have proven hypomagnesemia and the presence of its complications. PMID- 16690177 TI - Leptin and insulin response to long-term total parenteral nutrition depends on body fat mass. AB - BACKGROUND & AIMS: Circulating leptin and insulin concentrations are physiologically representing energy homeostasis. However, the artificial situation of long-term total parenteral nutrition (TPN) and its effects on serum leptin and insulin is not fully understood. METHODS: We studied 42 gastroenterological patients who received TPN for 19+/-11 days. Serum leptin and insulin levels as well as body composition assessed by bioelectrical impedance analysis were evaluated on days 0, 7 and 14. Insulin sensitivity was estimated by calculating the QUICKI. RESULTS: Before the start of TPN, leptin correlated positively with female gender (P<0.03), BMI (P<0.02), fat mass (P<0.02), insulin levels (P<0.001) and QUICKI (P<0.001). Within the first week of TPN, an increase of leptin levels was found only in patients with a body fat mass of >30% (P<0.02). As these were predominantly women, their leptin levels increased likewise (P<0.003). In regression analysis, fat mass (P<0.001), female gender (P<0.04), insulin levels (P<0.03), and i.v. glucose supply rates (P<0.05) were independently associated to leptin levels. CONCLUSIONS: TPN-especially glucose induces a neurohumoral response as shown here for leptin and insulin that is mainly depending on the fat mass. Better understanding of this regulatory mechanism during artificial nutrition could offer a new approach to improve its therapeutic effects. PMID- 16690178 TI - Cross-clade recognition and neutralization by the V3 region from clade C human immunodeficiency virus-1 envelope. AB - To understand the cross-reactivity of antibodies directed against variable regions of human immunodeficiency virus-1 (HIV-1) envelope (Env), chimeric immunogens were prepared from different clades with modifications in variable regions, and the resulting neutralizing antibody response was evaluated. The V3 specific neutralization activity induced by a clade B immunogen was limited to clade B viruses and was blocked by a clade B V3 peptide, but not by analogous clade A or C V3 peptides. In contrast, the V3 response elicited by a clade C immunogen cross-reacted with sensitive clade B viruses. The V3 region from a clade C virus, when introduced into a clade B sequence, elicited cross-clade activity, which could be reversed by V3 peptides derived from clades A and C. Thus, the anti-V3 antibody response elicited by a clade C immunogen could cross react with heterologous clade viruses. Additionally, we describe a V1-specific immune response that mediated neutralization limited to the homologous HIV IIIB isolate and may be partially responsible for the commonly observed strain specific neutralization responses elicited by vaccine immunogens. PMID- 16690179 TI - Insights from Australian parents into educational experiences in the early postnatal period. AB - OBJECTIVE: to investigate the provision of parent education during the early postnatal period in order to gain insight that, through stakeholder collaboration, will contribute to the development of innovative strategies to enhance the provision of postnatal education in a contemporary health-care environment. DESIGN: the study comprises the first stage of an action-research project. The first stage of research sought to explore the experiences of mothers and fathers in the early postnatal period by conducting a questionnaire within 4 weeks of the birth of their baby. The data obtained from the questionnaire is to inform an action-research group for stage two of the project. SETTING: The Children, Youth and Women's Health Service, a large city maternity hospital in South Australia, covering a range of socio-economic strata. PARTICIPANTS: 85 parents completed and returned the questionnaire, comprising 52 mothers and 33 fathers. MEASUREMENT: an anonymous self-report questionnaire was purpose designed to provide each parent with an opportunity to reflect on their own experience, with particular emphasis given to the provision of education and support during the early postnatal period. FINDINGS: a number of themes emerged, including a window of opportunity during the postnatal hospital stay to provide education and support, despite the reduction in the length of stay; the need for a family centred approach to maternity services; and the significance of self and social network in the early transition to parenthood. CONCLUSIONS: The findings from this stage of the research, combined with a review of the literature, provide insight that will contribute to stage two of the study. At this stage, an action research group will continue planning to develop specific actions to enhance the provision of education to parents in the early postnatal period. These actions will subsequently be implemented and assessed. PMID- 16690180 TI - The effects of Scaphoid and Colles casts on hand function. AB - Although necessary for bone healing, immobilisation temporarily prevents hand function and may necessitate corrective physiotherapy later. Scaphoid and Colles casts are both commonly used to immobilize scaphoid fractures. Non-union rates are comparable with both casts. The Scaphoid cast incorporates the thumb, whereas the Colles cast leaves the thumb free. We compared the effect of the two casts on hand function in 20 healthy right-hand-dominant volunteers using the Jebsen Taylor Hand Function Test. Data were obtained through a mixed between and within subject design. Both casts prolonged the time taken to complete the hand function test compared to controls. Testing in the Scaphoid cast took significantly longer than in the Colles cast. PMID- 16690181 TI - Can sarcoidosis cause autoamputation of a finger phalanx? AB - Sarcoidosis is a multisystemic granulomatous disease. In the case presented, autoamputation of the distal phalanx of a little finger of the left hand was observed. The possibility of autodigital amputation as a result of sarcoidosis is discussed. PMID- 16690182 TI - Development and validation of a behavioural assay to measure the tolerance of Hediste diversicolor to copper. AB - The behaviour of Hediste diversicolor from the Humber was investigated under different concentrations of copper sulphate. A range of behaviours were indicative of metal-stress. These included consistent attempts at burrowing, eversion of the proboscis and abnormal crawling. The bioassay itself consisted of exposing worms to increasing concentrations of copper sulphate and recording the concentration at which a stress response was elicited. The behavioural end-points were shown to be a good predictor of time of death of Fal estuary worms under acutely toxic conditions. The bioassay would therefore allow the separation of tolerance phenotypes without mortality to the worm. Worms were not affected by consecutive bioassays and it was proposed that tolerance to more than one metal could be determined for individual worms. PMID- 16690183 TI - Modelling 2,4-dichlorophenol bioavailability and bioaccumulation by the freshwater fingernail clam Sphaerium corneum using artificial particles and humic acids. AB - The complex and variable composition of natural sediments makes it very difficult to predict the bioavailability and bioaccumulation of sediment-bound contaminants. Several approaches have been proposed to overcome this problem, including an experimental model using artificial particles with or without humic acids as a source of organic matter. For this work, we have applied this experimental model, and also a sample of a natural sediment, to investigate the uptake and bioaccumulation of 2,4-dichlorophenol (2,4-DCP) by Sphaerium corneum. Additionally, the particle-water partition coefficients (K(d)) were calculated. The results showed that the bioaccumulation of 2,4-DCP by clams did not depend solely on the levels of chemical dissolved, but also on the amount sorbed onto the particles and the characteristics and the strength of that binding. This study confirms the value of using artificial particles as a suitable experimental model for assessing the fate of sediment-bound contaminants. PMID- 16690184 TI - A root cause analysis of clinical error: confronting the disjunction between formal rules and situated clinical activity. AB - This paper presents evidence from a root cause analysis (RCA) team meeting that was recently conducted in a Sydney Metropolitan Teaching Hospital to investigate an iatrogenic morphine overdose. Analysis of the meeting transcript reveals on three levels that clinical members of the team struggle with framing the uncertain and contradictory details of situated clinical activity and translating these first into 'root causes', and then into recommendations for practice change. This analysis puts two challenges into special relief. First, RCA team members find themselves in the unusual position of having to derive organizational-managerial generalizations from the specifics of in situ activity. Second, they are constrained by the expectation inscribed into RCA that their recommendations result in 'systems improvements' assumed to flow forth from an extension of formal rules and spread of procedures. We argue that this perspective misrecognizes the importance of RCA as a means to engender solutions that leave the procedural detail of clinical processes unspecified, and produce cross-hospital discussions about the organizational dimensions of care. PMID- 16690185 TI - The problems of offenders with mental disorders: a plurality of perspectives within a single mental health care organisation. AB - Managers, doctors, nurses, occupational therapists, social workers, psychologists, unqualified staff and service users were interviewed for a qualitative study of risk management and rehabilitation in an inner city medium secure forensic mental health care unit. Different professional orientations to service user problems were identified. Doctors focused primarily on the diagnosis of mental disorder, which they managed mainly through pharmaceutical interventions. Psychologists were principally concerned with personal factors, for example service user insight into their biographical history. Occupational therapists concentrated mainly on daily living skills, and social workers on post discharge living arrangements. Some front line nurses, held accountable for security lapses, adopted a criminogenic approach. Service users were more likely than professionals to understand their needs in terms of their wider life circumstances. These differences are explored qualitatively in relation to four models of crossdisciplinary relationships: monoprofessional self-organisation combined with restricted communication; hermeneutic reaching out to other perspectives; the establishment of interdisciplinary sub-systems; and transdisciplinary merger. Relationships between professions working in this unit, as portrayed in qualitative interviews, corresponded mainly to the first model of monoprofessional self-organisation. Reasons for restricted crossdisciplinary understanding, particularly the wide power/status differences between the medical and other professions, and between staff and patients, are discussed. PMID- 16690186 TI - Multiple measures of socioeconomic circumstances and common mental disorders. AB - While serious mental disorders typically show socioeconomic differences similar to physical illness-that is, that lower positions imply poorer health-differences for common mental disorders have been inconsistent. We aim to clarify the associations and pathways between measures of socioeconomic circumstances and common mental disorders by simultaneously analysing several past and present socioeconomic measures. The data were derived from middle-aged women and men employed by the City of Helsinki. Cross-sectional surveys were conducted in 2000 2002 among employees who, during each year, reached 40, 45, 50, 55 or 60 years of age. The pooled data include 8970 respondents (80% women; response rate 67%). Common mental disorders were measured by GHQ-12 and the SF-36 mental component summary. Seven socioeconomic measures were included: parental education, childhood economic difficulties, own education, occupational class, household income, home ownership, and current economic difficulties. Logistic regression analysis was used to examine associations between the socioeconomic circumstances and common mental disorders. Past and present economic difficulties were strongly associated with common mental disorders, whereas conventional past and present socioeconomic status measures showed weak or slightly reverse associations. Adjusting for age and gradually for each socioeconomic measure did not affect the main findings, which were very similar for women and men, as well as for both measures of common mental disorders. While the associations of conventional socioeconomic status measures with common mental disorders were weak and inconsistent, our results highlight the importance of past and present economic difficulties to these disorders. PMID- 16690187 TI - Psychosocial job characteristics and risk of mortality in a Japanese community based working population: the Jichi Medical School Cohort Study. AB - We prospectively investigated the association between psychosocial job characteristics according to the job demand-control model and the risk of mortality in a Japanese community-based working population. A baseline examination conducted from 1992 to 1995 determined the socioeconomic, behavioural, and biological risks in addition to the psychosocial job characteristics of 3178 male and 3331 female workers aged 65 and under and free from cancer and cardiovascular diseases. During the 9-year follow-up study, 157 men and 64 women died. In the follow-up, the results of Cox proportional hazards regression analysis revealed that men with concurrent high job demands and high job control (an active job) had the lowest risk of mortality from all causes. Compared with the low demand and high control job category, the multivariate relative risk of an active job was 0.53 (95% confidence interval: 0.31, 0.89). This finding appeared largely attributable to a reduction in cancer mortality. Job characteristics were not associated with cardiovascular diseases or external causes of mortality. For women, no significant associations were observed. The findings suggest that an active job has a beneficial effect on the health of Japanese male workers. Investigating of the effect of psychosocial job characteristics on cancer might therefore provide valuable insights into the health of workers. PMID- 16690188 TI - Altered coordination of the neuroendocrine response during psychosocial stress in subjects with high trait anxiety. AB - Contradicting data are available on stress responsiveness in subjects with high anxiety. In the present study we tested the hypothesis that high trait anxiety is associated with impaired coordination of the stress response, rather than global hypo- or hyper-responsiveness. The sample consisted of subjects with high (n=15) and with low (n=12) trait anxiety. Subjects with middle-range levels of anxiety were excluded from the study. After psychological characterization, the volunteers were exposed to a public speech procedure. A spectrum of neuroendocrine parameters was measured before, during and after the procedure and the results were analyzed by exploratory statistics. Psychological characterization of subjects revealed a lower preference for task-oriented but a higher one for emotion-oriented coping strategies as well as lower scores on hardiness in subjects with high trait anxiety. After the speech procedure, differences in selected mood and personality characteristics were observed, with the anxious group scoring significantly higher in scales for stress, tiredness, arousal, anxiety and depression. Factor analysis revealed that one common factor grouped blood pressure, catecholamine concentrations in blood and heart rate in non-anxious subjects, while three distinct factors separated these parameters in anxious subjects. Correlation analysis in anxious subjects showed that lower adrenocorticotropin (ACTH) and cortisol responses during stress were associated with exaggerated perception of stress and worse mental performance. Our findings indicate that subjects with high anxiety have different relationships between specific neuroendocrine parameters, subjective perception of stress and Stroop test performance. PMID- 16690189 TI - ESR and NMR spectroscopy studies on protein oxidation and formation of dityrosine in emulsions containing oxidised methyl linoleate. AB - Oxidised lipids are reported to interact with proteins causing undesirable changes in nutritional and functional properties including a loss of amino acids, cross-linking and damage to proteins and DNA. ESR spectroscopy with spin trapping was used to study the type of radical species generated in methyl linoleate and the transfer of the radical to protein beta-lactoglobulin. Antioxidants vitamins C and E reduced lipid oxidation and subsequent transfer of the radical to the protein as shown by the shape and size of the radical adduct. Changes to protein molecular structure due to oxidation were investigated by multidimensional NMR spectroscopy and liquid chromatography. NMR spectra indicated that as a result of oxidation and protein denaturation, there was an increase in structural flexibility and some initially protected backbone amide groups were exposed as they become sharper and easily identifiable. Dityrosine was detected in all samples tested which is indicative of oxidative damage to proteins. Monitoring tyrosyl radicals and formation of dityrosine is of practical value in order to enhance the acceptability, nutritional and safety aspects of proteins. PMID- 16690190 TI - Antidiabetic effect of garlic oil but not diallyl disulfide in rats with streptozotocin-induced diabetes. AB - We investigated the effects of garlic oil and diallyl disulfide (DADS) on glycemic control and renal function in rats with streptozotocin-induced diabetes. Rats received by gavage garlic oil (100 mg/kg body wt) or DADS (40 or 80 mg/kg body wt) every other day until 16 weeks after the induction of diabetes. The control rats were treated with corn oil only. Neither garlic oil nor DADS significantly affected fasting blood glucose concentrations throughout the investigation period. Garlic oil did not affect oral glucose tolerance in diabetes acutely but significantly improved oral glucose tolerance at 4, 8, 12, and 16 weeks and significantly ameliorated proteinuria at the end of 16 weeks. DADS did not significantly affect oral glucose tolerance or renal function. Diabetic rats fed 80 mg DADS/kg body wt had a significantly lower rate of body weight gain and a significantly lower ratio of muscle weight to body weight than did vehicle-treated diabetic rats. In conclusion, long-term treatment of diabetes with garlic oil can improve oral glucose tolerance and renal function in diabetes but not through the action of DADS. High doses of DADS may further complicate the metabolic disturbances in diabetes. PMID- 16690191 TI - [Liposarcoma of the thumb]. AB - Liposarcoma is the most common malignant deep soft-tissue tumor in adults. however, primitive cutaneous liposarcoma is very rare. We report the observation of a patient who presents a liposarcoma of the thumb. PMID- 16690192 TI - Amelioration of fumonisin B1 hepatotoxicity in mice by depletion of T cells with anti-Thy-1.2. AB - Fumonisin B1 is a mycotoxin produced by Fusarium verticillioides, frequently associated with corn. It produces species-specific and organ-specific toxicity, including equine leukoencephalomalacia, porcine pulmonary edema, and hepatic or renal damage in most animal species. Fumonisin B1 perturbs sphingolipid metabolism by inhibiting ceramide synthase. Our previous studies in male mice indicated that fumonisin B1-induced hepatotoxicity is modulated by the localized activation of cytokines in liver macrophages and other cell types. In the current study, male athymic nude mice and their wild type counterparts (WT), the latter with or without depletion of T cells, were treated subcutaneously with fumonisin B1 at 2.25 mg/kg/day for 5 days and sampled 24 h after the last injection. Depletion of T cells in WT was achieved by a single intravenous injection of 50 microg monoclonal antibody against Thy-1.2 surface antigen of mature peripheral T lymphocytes 24 h before the first fumonisin B1 treatment. The depletion of T cells nearly abolished fumonisin B1-mediated liver toxicity as indicated by the near normal concentrations of circulating liver enzymes and by enumeration of apoptotic hepatocytes. There was no difference in the fumonisin B1-induced elevation in circulating liver enzymes between WT and nude mice. Fumonisin B1 induced mRNA expression of tumor necrosis factor alpha and interleukin-1alpha was observed in nude and WT mice but not in T cell-depleted mice. Hepatotoxic response to fumonisin B1 was unaltered in mice lacking natural killer cells. This study suggested that T cells and corresponding proinflammatory cytokines have a vital role in mediating fumonisin B1-induced hepatic toxicity. PMID- 16690193 TI - Mechanisms underlying the anti-androgenic effects of diethylhexyl phthalate in fetal rat testis. AB - Diethylhexyl phthalate (DEHP) is widely used as a plasticizer in consumer products and is known to disturb the development of the male reproductive system in rats. The mechanisms by which DEHP exerts these effects are not yet fully elucidated, though some of the effects are related to reduced fetal testosterone production. The present study investigated the effects of four different doses of DEHP on fetal testicular histopathology, testosterone production and expression of proteins and genes involved in steroid synthesis in fetal testes. Pregnant Wistar rats were gavaged from GD 7 to 21 with vehicle, 10, 30, 100 or 300 mg/kg bw/day of DEHP. In male fetuses examined at GD 21, testicular testosterone production ex vivo and testicular testosterone levels were reduced significantly at the highest dose. Histopathological effects on gonocytes were observed at 100 and 300 mg/kg bw/day, whereas Leydig cell effects were mainly seen at 300 mg/kg bw/day. Quantitative RT-PCR revealed reduced testicular mRNA expression of the steroidogenesis related factors SR-B1, StAR, PBR and P450scc. Additionally, we observed reduced mRNA expression of the nuclear receptor SF-1, which regulates certain steps in steroid synthesis, and reduced expression of the cryptorchidism associated Insl-3. Immunohistochemistry showed clear reductions of StAR, PBR, P450scc and PPARgamma protein levels in fetal Leydig cells, indicating that DEHP affects regulation of certain steps in cholesterol transport and steroid synthesis. The suppression of testosterone levels observed in phthalate-exposed fetal rats was likely caused by the low expression of these receptors and enzymes involved in steroidogenesis. It is conceivable that the observed effects of DEHP on the expression of nuclear receptors SF-1 and PPARgamma are involved in the downregulation of steroidogenic factors and testosterone levels and thereby underlie the disturbed development of the male reproductive system. PMID- 16690194 TI - Analysis of purified gp96 preparations from rat and mouse livers using 2-D gel electrophoresis and tandem mass spectrometry. AB - The stress protein gp96 exhibits a number of immunological activities, the majority of studies into which have used gp96 purified from a variety of tissues. On the basis of 1-D gel electrophoresis, the purity of these preparations has been reported to range between 70% and 99%. This study analyzed gp96 preparations from rat and mouse livers using 2-D gel electrophoresis and liquid chromatography electrospray ionization tandem mass spectrometry (MS-MS). The procedure for purifying gp96 was reproducible, as similar protein profiles were observed in replicate gels of gp96 preparations. The purity of the preparations was typically around 70%, with minor co-purified proteins of varying molecular weights and mobilities being present. Dominant bands at 95-100 kDa in preparations from Wistar rats and C57BL/6 mice were identified as gp96 by ECL Western blotting. Multiple bands having similar, yet distinct molecular weights and differing pI mobility on ECL Western blots were confirmed as being gp96 in preparations from Wistar rats using MS-MS. The most striking feature of the 2-D gel analysis was the presence of additional dominant bands at 55 kDa in preparations from Wistar rats, and at 75-90 kDa in preparations from C57BL/6 mice. These were identified as gp96 by ECL Western blotting and, in the case of preparations from Wistar rats, by MS-MS. Although the lower molecular weight, gp96-related molecules might be partially degraded gp96, their reproducible presence, definition and characteristics suggest that they are alternative, species-specific isoforms of the molecule. A 55 kDa protein which exhibited a lower pI value than gp96 was present in all preparations and this was identified as calreticulin, another putative immunoregulatory molecule. This study confirms the reproducibility of the gp96 purification protocol and reveals the presence of multiple gp96 isoforms, some of which likely result from post-translational modifications such as differential glycosylation and phosphorylation. PMID- 16690195 TI - A genetic model to investigate drug-target interactions at the ribosomal decoding site. AB - Recent advances in X-ray crystallography have greatly contributed to the understanding of the structural interactions between aminoglycosides and the ribosomal decoding site. Efforts to genetically probe the functional relevance of proposed drug-nucleotide contacts have in part been hampered by the presence of multiple rRNA operons in most bacteria. A derivative of the Gram-positive Mycobacterium smegmatis was rendered single rRNA operon allelic by means of gene inactivation techniques. In this system, genetic manipulation of the single chromosomal rRNA operon results in cells carrying homogeneous populations of mutant ribosomes. An exhaustive mutagenesis study of the ribosomal A site has been performed to define the importance of individual drug-nucleotide contacts. Mutational alterations in the M. smegmatis decoding site are discussed here, comparing the results with those obtained in other organisms. Implications for the selectivity of antimicrobial agents and for the fitness cost of resistance mutations are addressed. PMID- 16690196 TI - Cyclic GMP modulates store-operated calcium entry inducing phosphatidylserine translocation at the surface of megakaryocytic cells. AB - When subjected to stimulation, cells from the vascular compartment show a spontaneous collapse of the plasma membrane phospholipid asymmetry and phosphatidylserine is exposed at the external leaflet. Thus, phosphatidylserine externalization is essential for normal hemostasis and phagocytosis. The mechanism governing the migration of phosphatidylserine to the exoplasmic leaflet is not yet fully understood. We have proposed that store-operated calcium entry (SOCE) constitutes a key step of this process. Here, interaction of [Ca(2+)](i), cAMP and cGMP pathways and phosphatidylserine exposure was examined in human megakaryocytic cells. The membrane permeable cAMP and cGMP analogues, pCPT-cAMP and pCPT-cGMP, enhanced the Ca(2+) signal induced by ionophore and SOCE. Responses to pCPT-cAMP and pCPT-cGMP were independent of protein kinase A, protein kinase G (PKG) or ERK pathways. Inhibition of small G-proteins reduced or abolished the increase of [Ca(2+)](i) induced by pCPT-cAMP or pCPT-cGMP, respectively. pCPT-cGMP but not pCPT-cAMP enhanced the ability of cells to expose phosphatidylserine. This effect was not prevented by the inhibition of PKG or small G-proteins. These results show the differential role of cyclic nucleotides in the Ca(2+)-dependent membrane remodeling. Hence, pCPT-cGMP is another regulatory element for the completion of SOCE-induced phosphatidylserine transmembrane redistribution in HEL cells through a mechanism implicating small G proteins. PMID- 16690197 TI - Crystal structures of GMP kinase in complex with ganciclovir monophosphate and Ap5G. AB - Guanosine monophosphate kinases (GMPK), by catalyzing the phosphorylation of GMP or dGMP, are of dual potential in assisting the activation of anti-viral prodrugs or as candidates for antibiotic strategies. Human GMPK is an obligate step for the activation of acyclic guanosine analogs, such as ganciclovir, which necessitate efficient phosphorylation, while GMPK from bacterial pathogens, in which this enzyme is essential, are potential targets for therapeutic inhibition. Here we analyze these two aspects of GMPK activity with the crystal structures of Escherichia coli GMPK in complex with ganciclovir-monophosphate (GCV-MP) and with a bi-substrate inhibitor, Ap5G. GCV-MP binds as GMP to the GMP-binding domain, which is identical in E. coli and human GMPKs, but unlike the natural substrate fails to stabilize the closed, catalytically-competent conformation of this domain. Comparison with GMP- and GDP-bound GMPK structures identifies the 2'hydroxyl of the ribose moiety as responsible for hooking the GMP-binding domain onto the CORE domain. Absence of this hydroxyl in GCV-MP impairs the stabilization of the active conformation, and explains why GCV-MP is phosphorylated less efficiently than GMP, but as efficiently as dGMP. In contrast, Ap5G is an efficient inhibitor of GMPK. The crystal structure shows that Ap5G locks an incompletely closed conformation of the enzyme, in which the adenine moiety is located outside its expected binding site. Instead, it binds at a subunit interface that is unique to the bacterial enzyme, which is in equilibrium between a dimeric and an hexameric form in solution. This suggests that inhibitors could be designed to bind at this interface such as to prevent nucleotide-induced domain closure. Altogether, these complexes point to domain motions as critical components to be evaluated in therapeutic strategies targeting NMP kinases, with opposite effects depending on whether efficient phosphorylation or inhibition is being sought after. PMID- 16690198 TI - Length and pH-dependent energetics of (CCG)n and (CGG)n trinucleotide repeats. AB - Trinucleotide repeats are involved in a number of debilitating diseases such as fragile-X syndrome and myotonic dystrophy. Eighteen to 75 base-long (CCG)(n) and (CGG)(n) oligodeoxynucleotides were analysed using a combination of biophysical (UV-absorbance, differential scanning calorimetry) and biochemical methods (non denaturing gel electrophoresis, enzymatic footprinting). All oligomers formed stable intramolecular structures under near physiological conditions with a melting temperature which was only weakly dependent on oligomer length. Thermodynamic analysis of the denaturation process by UV-melting and calorimetric experiments revealed a length-dependent discrepancy between the enthalpy values deduced from model-dependent (UV-melting) and model-independent experiments (calorimetry), as recently shown for CTG and CAG trinucleotides (Nucleic Acids Res. 33 (2005) 4065). Evidence for non-zero molar heat capacity changes was also derived from the analysis of the Arrhenius plots. Such behaviour is analysed in the framework of an intramolecular "branched" or "broken" hairpin model, in which long oligomers do not fold into a simple long hairpin-stem intramolecular structure, but allow the formation of several independent folding units of unequal stability. These results suggest that this observation may be extended to various trinucleotide repeats-containing sequences. PMID- 16690199 TI - Flavonoids inhibit the amidolytic activity of human thrombin. AB - The effect of a group of natural flavonoids on human thrombin amidolytic activity was investigated using a spectrophotometric inhibition assay while information on the kinetics and thermodynamics was obtained using optical biosensor techniques. All the flavonoids tested acted as reversible inhibitors, and the quercetin thrombin complex was found to be most stable at pH=7.5. Docking analysis indicated that quercetin's inhibitory behavior could be related to its planar structure and low steric hindrance, and to its ability to form a critical H-bond with thrombin His57. PMID- 16690200 TI - The increased accumulation of structurally modified versican and decorin is related with the progression of laryngeal cancer. AB - Versican and decorin, two proteoglycans (PGs) with contradictory roles in the pathophysiology of cancer, comprise important stromal components in many tumor types and play a crucial role in the progression of cancer. In this study, we provide direct evidence for a significant and stage-related accumulation of versican and decorin in the tumor-associated stroma of laryngeal squamous cell carcinoma (LSCC) in comparison to normal larynx. Both PGs were found to be co localized within the peritumorous stroma. In addition, the accumulated versican and decorin were markedly modified on both protein core and glycosaminoglycan (GAG) levels. Decorin, which was present under both glycanated and non-glycanated forms, perceptibly increased with the progression of LSCC, compared to the normal larynx. Tumor-associated glycanated decorin was found to contain significant amounts of dermatan sulfate (DS) sequences. Versican was also found to undergo stage-related structural modifications since a marked heterogeneity of protein cores was observed, being intense in late stage of laryngeal cancer. The increased accumulation of both versican and decorin was associated with a significant stage-related increase of the molar ratio of Delta di-mono4S to Delta di-mono6S up to approximately threefold in LSCC compared to the normal ones. The modified chemical structure of both PGs could be associated with the degree of aggressiveness of laryngeal squamous cell carcinomas. PMID- 16690201 TI - Viscosity B-coefficients and standard partial molar volumes of amino acids, and their roles in interpreting the protein (enzyme) stabilization. AB - This review systematically surveys the viscosity B-coefficients and standard partial molar volumes of amino acids at various temperatures as these data are quite important for interpreting the hydration and other properties of peptides and proteins. The effect of organic solutes and various ions on the viscometric and volumetric properties of amino acids has also been discussed in terms of their kosmotropic ('structure-making') effects on the hydration of amino acids. The comparison of these effects on the amino acid hydration enables us to have a better understanding of the influence of organic solute and salt on the protein stabilization. In addition, the viscometric and volumetric behaviors of amino acid ions (cations and anions) are also summarized because these ions have recently been incorporated as part of novel ionic liquids, which have wide applications in biocatalysis and protein stabilization. PMID- 16690202 TI - Sequestration of biogenic amines by alginic and fulvic acids. AB - The interaction of natural (alginic and fulvic acids) and synthetic (polyacrylic acid 2.0 kDa) polyelectrolytes with some protonated polyamines [diamines: ethylendiamine, 1,4-diaminobutane (or putrescine), 1,5-diaminopentane (or cadaverine); triamines: N-(3-aminopropyl)-1,4-diaminobutane (or spermidine), diethylenetriamine; tetramine: N,N'-bis(3-aminopropyl)-1,4-diaminobutane (or spermine); pentamine: tetraethylene-pentamine; hexamine: pentaethylenehexamine] was studied at T=25 degrees C by potentiometry and calorimetry. Measurements were performed without supporting electrolyte, in order to avoid interference, and results were reported at I=0 mol L(-)(1). For all the systems, the formation of (am)L(2)H(i) species was found (am=amine; L=polyelectrolyte; i=1...4, depending on the amine considered). The stability of polyanion-polyammonium cation complexes is always significant, and for high-charged polycations, we observe a stability comparable to that of strong metal complexes. For example, by considering the formation reaction (am)H(i)+2L=(am)L(2)H(i) we found log K(i)=6.0, 6.5 and 10.8 for i=1, 2 and 3, respectively, in the system alginate spermidine. Low and positive formation DeltaH(degrees) values indicate that the main contribution to the stability is entropic in nature. The sequestering ability of polyelectrolytes toward amines was modelled by a sigmoid Boltzman type equation. Some empirical relationships between stability, charges and DeltaG(degrees) and TDeltaS(degrees) are reported. Mean values per salt bridge of formation thermodynamic parameters (DeltaX(degrees) (n)) are DeltaG(degrees) (n)= 5.8+/-0.4, DeltaH degrees (n)=0.7+/-0.5 and TDeltaS(degrees) (n)=6.5+/-0.5 kJmol( )(1) for all the systems studied in this work. PMID- 16690203 TI - A quantitative method for zoning of protected areas and its spatial ecological implications. AB - Zoning is a key prescriptive tool for administration and management of protected areas. However, the lack of zoning is common for most protected areas in developing countries and, as a consequence, many protected areas are not effective in achieving the goals for which they were created. In this work, we introduce a quantitative method to expeditiously zone protected areas and we evaluate its ecological implications on hypothetical zoning cases. A real-world application is reported for the Talampaya National Park, a UNESCO World Heritage Site located in Argentina. Our method is a modification of the zoning forest model developed by Bos [Bos, J., 1993. Zoning in forest management: a quadratic assignment problem solved by simulated annealing. Journal of Environmental Management 37, 127-145.]. Main innovations involve a quadratic function of distance between land units, non-reciprocal weights for adjacent land uses (mathematically represented by a non-symmetric matrix), and the possibility of imposing a connectivity constraint. Due to its intrinsic spatial dimension, the zoning problem belongs to the NP-hard class, i.e. a solution can only be obtained in non-polynomial time [Nemhausser, G., Wolsey, L., 1988. Integer and Combinatorial Optimization. John Wiley, New York.]. For that purpose, we applied a simulated annealing heuristic implemented as a FORTRAN language routine. Our innovations were effective in achieving zoning designs more compatible with biological diversity protection. The quadratic distance term facilitated the delineation of core zones for elements of significance; the connectivity constraint minimized fragmentation; non-reciprocal land use weightings contributed to better representing management decisions, and influenced mainly the edge and shape of zones. This quantitative method can assist the zoning process within protected areas by offering many zonation scheme alternatives with minimum cost, time and effort. This ability provides a new tool to improve zoning within protected areas in developing countries. PMID- 16690204 TI - Dynamic modeling of chemical fate and transport in multimedia environments at watershed scale-I: theoretical considerations and model implementation. AB - A geo-referenced environmental fate model was developed for analyzing unsteady state dispersion and distribution of chemicals in multimedia environmental systems. Chemical transport processes were formulated in seven environmental compartments of air, canopy, surface soil, root-zone soil, vadose-zone soil, surface water, and sediment. The model assumed that the compartments were completely mixed and chemical equilibrium was established instantaneously between the sub-compartments within each compartment. A fugacity approach was utilized to formulate the mechanisms of diffusion, advection, physical interfacial transport, and transformation reactions. The governing equations of chemical mass balances in the environmental compartments were solved simultaneously to reflect the interactions between the compartments. A geographic information system (GIS) database and geospatial analysis were integrated into the chemical transport simulation to provide spatially explicit estimations of model parameters at watershed scale. Temporal variations of the environmental properties and source emissions were also considered in the parameter estimations. The outputs of the model included time-dependent chemical concentrations in each compartment and its sub-compartments, and inter-media mass fluxes between adjacent compartments at daily time steps. PMID- 16690205 TI - Human embryonic stem cells as a cellular model for human disorders. AB - Human embryonic stem cells (HESCs) are pluripotent cell lines derived from the inner cell mass (ICM) of embryos at the blastocyst stage. These cells possess self renewal capacity and differentiation potential to all three embryonic germ layers. These unique characters made HESCs an attractive research tool for studying early human developmental processes as well as a potential therapeutic tool for various human diseases. Here, we focus on HESCs as a cellular model for human disorders. The advantages of such models as well as the various methodologies to achieve HESCs carrying a genetic defect will be discussed. PMID- 16690206 TI - Sensitive Ewing sarcoma and neuroblastoma cell lines have increased levels of BAD expression and decreased levels of BAR expression compared to resistant cell lines. AB - The purpose of the study is to compare the mRNA expression of pro-apoptotic and anti-apoptotic genes by using the GeneSystem 320 (Capital Genomix Inc., Gaithersburg, MD) to examine the differential expression in Ewing sarcoma and neuroblastoma cell lines. This is an alternate method for which internal controls have been built into the system for comparing mRNA. The tumor cell lines were chosen based on their previously characterized Fas-resistance or Fas-sensitive properties in order to determine the differences in their response to apoptotic signals. Two representative pro-apoptotic genes (BAD and SMAC) and one anti apoptotic gene (BAR) were chosen for the study. The results of mRNA expression were correlated with protein expression by Western analysis. BAD was highly expressed in the Fas-sensitive cell lines while SMAC was equally expressed in both Fas-sensitive and Fas-resistant cell lines. On the other hand, BAR was highly expressed in Fas-resistant cell lines and minimally expressed in the Fas sensitive cell lines. Our data suggests that levels of BAD and BAR mRNA expression predict sensitivity to apoptosis. PMID- 16690207 TI - Gene-environment interactions between smoking and a haplotype of RAI, ASE-1 and ERCC1 polymorphisms among women in relation to risk of lung cancer in a population-based study. AB - Homozygous carriers of a haplotype consisting of ERCC1 Asn118Asn(A), ASE-1 G 21A(G), RAI IVS1 A4364G(A) are at increased risk of lung cancer especially among women. Here, we analyse for gene-environment interactions with the predefined haplotype in a case cohort study including 428 lung cancer cases and a comparison group of 800 persons, all from the prospective Diet, Cancer and Health cohort of 57,000 Danes. At high smoking intensity (>20g tobacco/day), there was only additional risk of smoking intensity among women who were homozygous carriers of the haplotype (IRR=2.03; 95% CI: 1.10-3.73 per 5 additional g tobacco/day). PMID- 16690208 TI - Tic distribution and inhibitory processes in the sensorimotor circuit during adolescence: a cross-sectional TMS study. AB - Deficient inhibitory processes within the sensorimotor circuit, reflected by a shortened cortical silent period (CSP), have previously been described in both children and adults with tic disorders (TD) using transcranial magnetic stimulation (TMS). In contrast to adults, tic distribution (presence or absence of distal tics) did not affect CSP duration in children. The aim of this developmental TMS study was to clarify this striking difference. 127 children with TD were stratified into three age-groups (8-11.5, 11.5-15, 15-19 years) with and without distal tics. CSP was recorded from the abductor digiti minimi. Statistics revealed a significant tic distributionxage interaction effect. Only in the 15-19 years subgroup, CSP was shorter in patients with distal tics in comparison to patients without distal tics (94.1+/-54.1 ms versus 135.2+/-36.8 ms at a stimulus intensity of active motor threshold plus 30%). Inhibitory processes in the sensorimotor circuit could reflect developmental aspects of tic phenomenology, particularly tic distribution during adolescence. PMID- 16690210 TI - Expression of pannexin1 in the CNS of adult mouse: cellular localization and effect of 4-aminopyridine-induced seizures. AB - The expression pattern of pannexin1, a gene coding for a protein that forms gap junction channels, was studied as both mRNA and protein in the CNS of adult mouse. Pannexin1 was widely expressed in the CNS by neuronal cell types but not glial cells, except for Bergmann glial cells of the cerebellar cortex. Cells positive to Ca-binding proteins, principally parvalbumin, but also calbindin and calretinin, as well as glutamate decarboxylase 67 kDa isoform, were pannexin1 positive. Pannexin1 labeling was found in cells which are known to exhibit spontaneous and synchronous discharge, such as neurons of the inferior olivary complex and the reticular thalamic nucleus, and also in neurons whose electrical activity is not coupled with neighboring cells, such as motoneurons of the spinal cord. The analysis of cellular localization showed puncta that surrounded cell bodies (e.g. the pyramidal cells of hippocampus) or restricted areas inside the cell bodies (e.g. the spinal motoneurons). In Bergmann glial cells the staining was present as fine grains that covered a large part of the cellular surface. Pannexin1 stained cells that previous studies have reported as expressing connexin36, another protein forming gap junction channels. Thus, it was possible that these two proteins could be integrated in the same functions. Since connexin36 expression levels change after seizures, we examined the expression of both pannexin1 and connexin36 in cerebral cortex, hippocampus, cerebellum and brain stem at different time intervals (2, 4 and 8 h) after i.p. injection of 4 aminopyridine, which resulted in systemic seizures. The only modification of the expression levels observed in this study concerned the progressive decrement of the connexin36 in the hippocampus, while pannexin1 expression was unchanged. This finding suggested that pannexin1 and connexin36 are involved in different functional roles or that they are expressed in different cell types and that only those expressing the Cx36 are induced to apoptosis by epileptic seizures. PMID- 16690211 TI - The native T-type calcium current in relay neurons of the primate thalamus. AB - The generation of thalamic bursts depends upon calcium currents that flow through transiently open (T)-type calcium channels. In this study, we characterized the native T-type calcium current underlying thalamic burst responses in the macaque monkey. Current clamp recordings from lateral geniculate nucleus (LGN) slices showed characteristic burst responses when relay cells were depolarized from relatively hyperpolarized membrane potentials. These bursts could also be elicited by stimulation of excitatory synaptic inputs to LGN cells. Under voltage clamp conditions, the inactivation kinetics of native currents recorded from primate LGN neurons showed consistency with T-type currents recorded in other mammals and in expression systems. Real-time reverse transcriptase PCR performed on RNA isolated from the LGN (including tissues isolated from magnocellular and parvocellular laminae) detected voltage-dependent calcium channel (Ca(v)) 3.1, Ca(v) 3.2, and Ca(v) 3.3 channel transcripts. Ca(v) 3.1 occurred at relatively higher expression than other isoforms, consistent with in situ hybridization studies in rats, indicating that the molecular basis for burst firing in thalamocortical systems is an important conserved property of primate physiology. Since thalamic bursts have been observed during visual processing as well as in a number of CNS disorders, studies of the expression and modulation of these currents at multiple levels are critical for understanding their role in vision and for the discovery of new treatments for disruptions of thalamic rhythms. PMID- 16690213 TI - Early postnatal isolation reduces dopamine levels, elevates dopamine turnover and specifically disrupts prepulse inhibition in Nurr1-null heterozygous mice. AB - Sensorimotor gating is a phenomenon that is linked with dopamine neurotransmission in limbic and cortical areas, and disruption of sensorimotor gating has been consistently demonstrated in schizophrenia patients. The nuclear receptor Nurr1 is essential for development of dopamine neurons and, using Nurr1 null heterozygous mice, has been found to be important for normal dopamine neurotransmission as null heterozygous mice have reduced limbic and cortical dopamine levels and elevated open-field locomotor activity. The current investigation compared sensorimotor gating, as measured by prepulse inhibition of the acoustic startle response, in Nurr1 wild-type and null heterozygous mice. When mice were weaned between 19 and 21 days of age either into isolation or groups of three to five and tested 12 weeks later, prepulse inhibition was elevated in group-raised null heterozygous mice and significantly disrupted in isolated null heterozygous mice as compared with isolation-raised wild-type mice and group-raised null heterozygous mice. Isolation had no effect on prepulse inhibition in wild-type mice. Isolation reduced tissue dopamine levels and elevated dopamine turnover in the nucleus accumbens and striatum in both wild type and null heterozygous mice. In the prefrontal cortex, isolation reduced dopamine and 3,4-dihydroxyphenylacetic acid levels in null heterozygous as compared with isolation-raised wild-type mice, whereas no differences were observed between group-raised wild-type and null heterozygous mice. Neither the null heterozygous genotype nor isolation had any effect on basal or stress induced corticosterone levels. These data suggest that the Nurr1 null heterozygous genotype predisposes these mice to isolation-induced disruption of prepulse inhibition that may be related to the interactions between intrinsic deficiencies in dopamine neurotransmission as a result of the null heterozygous genotype and isolation-induced changes in dopamine neurotransmission. Post weaning isolation of Nurr1 null heterozygous mice provides a model to explore the interactions of genetic predisposition and environment/neurodevelopment on dopamine function that has important relevance to neuropsychiatric disorders. PMID- 16690212 TI - Gene expression in the rat cerebral cortex: comparison of recovery sleep and hypnotic-induced sleep. AB - Most hypnotic medications currently on the market target some aspect of GABAergic neurotransmission. Although all such compounds increase sleep, these drugs differentially affect the activity of the cerebral cortex as measured by the electroencephalogram. Whereas benzodiazepine medications such as triazolam tend to suppress slow wave activity in the cortex, the GABA(B) ligand gamma hydroxybutyrate greatly enhances slow wave activity and the non-benzodiazepine, zolpidem, which binds to the omega1 site on the GABA(A) receptor/Cl(-) ionophore complex, is intermediate in this regard. Our previous studies have demonstrated that a small number of genes exhibit increased expression in the cerebral cortex of the mouse and rat during recovery sleep after sleep deprivation: egr-3, fra-2, grp78, grp94, ngfi-b, and nr4a3. Using these genes as a panel of biomarkers associated with sleep, we asked whether hypnotic medications induce similar molecular changes in the rat cerebral cortex to those observed when both sleep continuity and slow wave activity are enhanced during recovery sleep. We find that, although each drug increases the expression of a subset of genes in the panel of biomarkers, no drug fully replicates the molecular changes in the cortex associated with recovery sleep. Furthermore, high levels of slow wave activity in the cortex are correlated with increased expression of fra-2 whereas the expression of grp94 is correlated with body temperature. These results demonstrate that sleep-related changes in gene expression may be affected by physiological covariates of sleep and wakefulness rather than by vigilance state per se. PMID- 16690214 TI - Maturation of firing pattern in chick vestibular nucleus neurons. AB - The principal cells of the chick tangential nucleus are vestibular nucleus neurons participating in the vestibuloocular and vestibulocollic reflexes. In birds and mammals, spontaneous and stimulus-evoked firing of action potentials is essential for vestibular nucleus neurons to generate mature vestibular reflex activity. The emergence of spike-firing pattern and the underlying ion channels were studied in morphologically-identified principal cells using whole-cell patch clamp recordings from brain slices of late-term embryos (embryonic day 16) and hatchling chickens (hatching day 1 and hatching day 5). Spontaneous spike activity emerged around the perinatal period, since at embryonic day 16 none of the principal cells generated spontaneous action potentials. However, at hatching day 1, 50% of the cells fired spontaneously (range, 3 to 32 spikes/s), which depended on synaptic transmission in most cells. By hatching day 5, 80% of the principal cells could fire action potentials spontaneously (range, 5 to 80 spikes/s), and this activity was independent of synaptic transmission and showed faster kinetics than at hatching day 1. Repetitive firing in response to depolarizing pulses appeared in the principal cells starting around embryonic day 16, when <20% of the neurons fired repetitively. However, almost 90% of the principal cells exhibited repetitive firing on depolarization at hatching day 1, and 100% by hatching day 5. From embryonic day 16 to hatching day 5, the gain for evoked spike firing increased almost 10-fold. At hatching day 5, a persistent sodium channel was essential for the generation of spontaneous spike activity, while a small conductance, calcium-dependent potassium current modulated both the spontaneous and evoked spike firing activity. Altogether, these in vitro studies showed that during the perinatal period, the principal cells switched from displaying no spontaneous spike activity at resting membrane potential and generating one spike on depolarization to the tonic firing of spontaneous and evoked action potentials. PMID- 16690215 TI - Delayed minocycline treatment reduces long-term functional deficits and histological injury in a rodent model of focal ischemia. AB - The absence of effective treatments for stroke presents a critical need for novel strategies that can reduce ischemic injury. Neuroinflammation following focal ischemia induces secondary injury in the region surrounding the insult, thus anti inflammatory agents are potential neuroprotectants. Minocycline is one such agent possessing neuroprotective properties, however many studies examining minocycline after ischemia have used minimal delays between ischemia and treatment, short survival periods, and lack measures of functional outcome. Such studies do not distinguish whether minocycline provides sustained protection or merely delays cell death. This study was designed to address some of these concerns. Male Sprague-Dawley rats were treated with multiple doses of minocycline (45 mg/kg i.p.) or vehicle beginning 2.5 h after endothelin-1-induced focal ischemia. Measures of forelimb asymmetry and skilled reaching (staircase test) were used to determine functional outcome 7, 15 and 28 days after ischemia. Long-term functional assessment indicates that minocycline provides limited benefit in the staircase test, but confers long-term benefit in the forelimb asymmetry test. Subcortical and whole hemisphere infarct volumes were reduced by 41 and 39% respectively in minocycline-treated animals. Further analysis revealed that minocycline attenuated long-term white matter damage adjacent to the striatal injury core, which correlated with sustained functional benefits. This study indicates that delayed minocycline treatment improves long-term functional outcome which is linked to protection of both white and gray matter. PMID- 16690216 TI - Ephrin-A5 modulates the topographic mapping and connectivity of commissural axons in murine hippocampus. AB - Entorhinal and commissural/associational projections show a non-overlapping distribution in the hippocampus proper and the dentate gyrus. The expression of Ephrins and their Eph receptors in the developing hippocampus indicates that this family of axonal guidance molecules may modulate the formation of these connections. Here we focused on the role of the ephrin-A5 ligand in the development of the main hippocampal afferents. In situ hybridization showed that ephrin-A5 mRNA was detected mainly in the principal cells of the hippocampus proper and in the dentate gyrus throughout postnatal development. Immunocytochemical analyses revealed prominent expression of the EphA3 receptor, a putative receptor for ephrin-A5, in the main cells and the neuropil of the developing hippocampus. Tracing experiments in ephrin-A5(-/-) mice showed that commissural projections were transiently altered in the hippocampus proper at P5, but they were mistargeted throughout the postnatal development in the dentate gyrus. Immunocytochemistry with anti-calbindin antibodies revealed that the dentate mossy fiber projection was not altered in ephrin-A5(-/-) mice. Electron microscopy studies showed alterations in the density of synapses and spines in commissural/associational layers, but not in entorhinal layers, and in the mossy fibers in these animals. Taken together, these findings indicate that ephrin-A5 signaling is involved in the formation and maturation of synapses in the hippocampus. PMID- 16690217 TI - Unique presynaptic and postsynaptic roles of Group II metabotropic glutamate receptors in the modulation of thalamic network activity. AB - The thalamic reticular nucleus (TRN) is a sheet of GABAergic neurons that project to other TRN neurons and to associated thalamocortical relay nuclei. The TRN receives glutamatergic synaptic inputs from cortex as well as reciprocal inputs from the collaterals of thalamocortical neurons. In addition to ionotropic glutamate receptors, metabotropic glutamate receptors (mGluRs) are present in the TRN circuitry. Using whole cell voltage clamp recordings, we pharmacologically characterized unique pre- and postsynaptic functions for Group II mGluRs (mGluR 2 and mGluR 3) within the TRN circuitry in ferrets. mGluR 2 was found on presynaptic cortical axon terminals in the TRN, where it reduced glutamate release, while mGluR 3 acted postsynaptically on TRN cells to increase membrane conductance. Using miniature inhibitory postsynaptic current analysis, we also found that picrotoxin-sensitive intra-TRN GABA-mediated neurotransmission was not affected by administration of a Group II mGluR agonist, indicating that neither mGluR 2 nor 3 acts on presynaptic GABA-containing terminals within the TRN. Because strong corticothalamic activation is implicated in abnormal thalamic rhythms, we used extracellular recordings in the lateral geniculate nucleus to study the effect of Group II mGluR agonists upon these slow oscillations. We induced approximately 3 Hz spike-and-wave discharge activity through corticothalamic stimulation, and found that such activity was reduced in the presence of the Group II mGluR agonist, (-)-2-oxa-4-aminobicyclo[3.1.0]hexane-4,6 dicarboxylate (LY379268). These data indicate that Group II mGluR reduce the impact of corticothalamic excitation, and that they may be a useful target in the reduction of absence-like rhythms. PMID- 16690218 TI - Inflammation increases the excitability of masseter muscle afferents. AB - Temporomandibular disorder is a major health problem associated with chronic orofacial pain in the masticatory muscles and/or temporomandibular joint. Evidence suggests that changes in primary afferents innervating the muscles of mastication may contribute to temporomandibular disorder. However, there has been little systematic study of the mechanisms controlling the excitability of these muscle afferents, nor their response to inflammation. In the present study, we tested the hypotheses that inflammation increases the excitability of sensory neurons innervating the masseter muscle of the rat and that the ionic mechanisms underlying these changes are unique to these neurons. We examined inflammation induced changes in the excitability of trigeminal ganglia muscle neurons following intramuscular injections of complete Freund's adjuvant. Three days after complete Freund's adjuvant injection acutely dissociated, retrogradely labeled trigeminal ganglia neurons were studied using whole cell patch clamp techniques. Complete Freund's adjuvant-induced inflammation was associated with an increase in neuronal excitability marked by a significant decrease in rheobase and increase in the slope of the stimulus response function assessed with depolarizing current injection. The increase in excitability was associated with significant decreases in the rate of action potential fall and the duration of the action potential afterhyperpolarization. These changes in excitability and action potential waveform were associated with significant shifts in the voltage dependence of activation and steady-state availability of voltage-gated K(+) current as well as significant decreases in the density of voltage-gated K(+) current subject to steady-state inactivation. These data suggest that K(+) channel subtypes may provide novel targets for the treatment of pain arising from inflamed muscle. These results also support the hypothesis that the underlying mechanisms of pain arising from specific regions of the body are unique suggesting that it may be possible, if not necessary to treat pain originating from different parts of the body with specific therapeutic interventions. PMID- 16690219 TI - Cetirizine as a novel weapon against reperfusion injury. PMID- 16690220 TI - A moat around castle walls. The role of axillary and facial hair in lymph node protection from mutagenic factors. AB - Axillary hair is a highly conserved phenotypical feature in humans, and as such deserves at least consideration of its functional significance. Protection from environmental factors is one of the main functions attributed to hair in furred vertebrates, but is believed to be inapplicable to humans. I considered the hypothesis that the phenotypic preservation of axillary hair is due to its unrecognized role in the organism protection. Two immediate questions arise--what exactly is being protected and what it is protected from. A large group of axillary lymph nodes represents a major difference between underarms and the adjacent areas of the trunk. The consideration of potential factors from which hair can offer protection identifies sunlight as the most likely candidate. Intense sweat production underarms may represent an independent defense mechanism, specifically protecting lymph nodes from overheating. Moreover, the pattern of facial hair growth in males strikingly overlaps with the distribution of superficial lymph nodes, suggesting potential role for facial hair in protection of lymph nodes, and possibly thymus and thyroid. The idea of lymph node protection from environmental mutagenic factors, such as UV radiation and heat, appears particularly important in light of wide association of lymph nodes with cancers. The position of contemporary fashion towards body hair is aggressively negative, including the social pressure for removal of axillary and bikini line hair for women, facial hair for men in many professional occupations, and even body hair for men. If this hypothesis is proven to be true, the implications will be significant for immunology (by providing new insights in lymph node physiology), health sciences (depilation is painful and therefore easily modifiable habit if proven to increase disease risk), as well as art, social fashion and economy. PMID- 16690221 TI - Epidermal growth factor receptor as a prognostic factor in locally advanced rectal-cancer patients treated with preoperative chemoradiation. AB - PURPOSE: We investigated the prognostic value of epidermal growth factor receptor (EGFR) expression in pretreatment biopsy specimens from patients with locally advanced rectal cancer treated with preoperative chemoradiation. METHODS AND MATERIALS: Pretreatment biopsy specimens from 92 patients with locally advanced rectal cancer were examined for EGFR expression by immunohistochemistry. EGFR expression was assessed by immunoreactive score (IRS). The prognostic value of EGFR expression was evaluated according to the level of EGFR expression. RESULTS: Epidermal growth factor receptor expression was positive in 65 patients (71%). EGFR expression levels were low (IRS 0 to 5) in 83 patients (90%) and high (IRS 6 to 7) in 9 patients (10%). A high level of EGFR expression was statistically significant for shorter overall survival (p = 0.013), disease-free survival (p = 0.002), and distant metastasis-free survival (p = 0.003), as compared with a low level of expression in univariate analysis. Grouping based on positive or negative EGFR expression did not represent prognostic significance for survival. In multivariate analysis, high EGFR expression was an independent prognostic factor for decreased disease-free survival (relative risk 2.4, p = 0.041) and distant metastasis-free survival (relative risk 2.6, p = 0.04). CONCLUSIONS: Our results suggest that high level of EGFR expression in a pretreatment biopsy specimen may be a significant adverse prognostic factor for disease-free survival and distant metastasis-free survival. PMID- 16690222 TI - Bauhinia candicans stimulation of glucose uptake in isolated gastric glands of normal and diabetic rabbits. AB - A butanol extract of dried leaves of Bahuinia candicans showed, in vitro, a stimulatory effect of glucose uptake in isolated gastric glands of normal and alloxan-diabetic rabbits. The extract (0.001-0.07 mg/mg protein) produced a dose dependent effect and was similar to the effect of insulin (1-100 nM). These findings suggest that the extract contains compound(s) that efficiently increase glucose transport at the basolateral side of isolated gastric glands. PMID- 16690223 TI - In vitro antifungal activities of Allium cepa, Allium sativum and ketoconazole against some pathogenic yeasts and dermatophytes. AB - By using an agar dilution assay, the antifungal activity of aqueous extracts prepared from Allium cepa (onion; AOE) and Allium sativum (garlic; AGE) were evaluated against Malassezia furfur (25 strains), Candida albicans (18 strains), other Candida sp. (12 strains) as well as 35 strains of various dermatophyte species and compared with the activity of a known antifungal drug, ketoconazole (KTZ). All the AOE, AGE and KTZ were found to be able to inhibit growth of all fungi tested in a dose-dependent manner with maximum of 100% at defined concentrations. The results indicate that onion and garlic might be promising in treatment of fungal-associated diseases from important pathogenic genera Candida, Malassezia and the dermatophytes. PMID- 16690224 TI - Antimicrobial activity of the Maniltoa schefferi extracts. AB - All fractions of Maniltoa schefferi exhibited a broad spectrum of antibacterial activity. Good activity was demonstrated by the petrol and dichloromethane fractions of seed pods, seeds with coat and by methanol fractions of stem heartwood, root bark, root heartwood and stem bark whose butanol fraction was equally good. None of the fractions showed any antifungal activity. PMID- 16690225 TI - Antimicrobial and antiplasmid activities of essential oils. AB - The antimicrobial and antiplasmid activities of essential oils (orange oil, eucalyptus oil, fennel oil, geranium oil, juniper oil, peppermint oil, rosemary oil, purified turpentine oil, thyme oil, Australian tea tree oil) and of menthol, the main component of peppermint oil, were investigated. The antimicrobial activities were determined on the Gram (+) Staphylococcus epidermidis and the Gram (-) Escherichia coli F'lac K12 LE140, and on two yeast Saccharomyces cerevisiae 0425 delta/1 and 0425 52C strains. The antiplasmid activities were investigated on E. coli F'lac bacterial strain. Each of the oils exhibited antimicrobial activity and three of them antiplasmid action. The interaction of peppermint oil and menthol with the antibiotics was studied on the same bacterial strain with the checkerboard method. Peppermint oil and menthol displayed additive synergy with oxytetracycline. A new mechanism of plasmid curing was established for one of the oil components. PMID- 16690226 TI - Antibacterial activity of Leonurus sibiricus aerial parts. AB - Different solvent extracts (carbon tetrachloride, chloroform, acetone and methanol) of Leonurus sibiricus were studied for their antibacterial activity. Carbon tetrachloride and chloroform extracts showed a broad spectrum of antibacterial activity. PMID- 16690227 TI - Screening for antioxidant properties of Allium giganteum. AB - The antioxidative properties of bulb, leaf and stalk of Allium giganteum were investigated. Activities of antioxidant enzymes (superoxide dismutase, catalase, peroxidase, glutathione peroxidase), quantities of malonyldialdehyde, superoxide and hydroxyl radicals and reduced glutathione and also the content of total flavonoids, chlorophylls a and b, carotenoids, vitamin C and soluble proteins were determined. The highest antioxidant activity was observed in the leaves. Furthermore, ESR signal of PBN-OH radical adducts in the presence of leaves phosphate buffer (pH 7) extract was reduced for 74.19%. PMID- 16690228 TI - [A cytological, immunophenotypical and cytogenetical study of 136 consecutive cases of B-cell chronic lymphoid hemopathies]. AB - A cytological, immunophenotypical and cytogenetical study of 136 chronic B-cell proliferations (93 CLL, 43 B-cell lymphomas) was led in order to precise diagnosis and to characterize and appreciate chromosomal rearrangements. In this series, mainly selected on blood lymphocytosis criteria, B-CLL were twice more frequent than small B-cell lymphomas. Probes used revealed cryptic abnormalities, which remained unknown by conventional cytogenetics (CC). The frequency of clonal abnormalities (CC and FISH) was 74.8% for this series, with 74.4% for lymphomas and 75.3% for CLL, mainly of Binet stage A (69 A, 13 B, 1 C, 10 unspecified). Proportion was 88.4% in A stages and 84.6% in B stages. In CLL, 13q14 cryptic deletions and translocations were widely majority, 14q32 translocations and trisomy 12 being predominant in lymphoma series. Interphase FISH study of non clonal metaphasic abnormalities with locus-specific probes often revealed unrecognised clones. PMID- 16690229 TI - Phenotype in X chromosome rearrangements: pitfalls of X inactivation study. AB - OBJECTIVE: X inactivation pattern in X chromosome rearrangements usually favor the less unbalanced cells. It is correlated to a normal phenotype, small size or infertility. We studied the correlation between phenotype and X inactivation ratio in patients with X structural anomalies. PATIENTS AND METHODS: During the 1999-2005 period, 12 X chromosome rearrangements, including three prenatal cases, were diagnosed in the Laboratoire de Cytogenetique of Strasbourg. In seven cases, X inactivation ratio could be assessed by late replication or methylation assay. RESULTS: In three of seven cases (del Xp, dup Xp, t(X;A)), X inactivation ratio and phenotype were consistent. The four other cases showed discrepancies between phenotype and X inactivation pattern: mental retardation and dysmorphism in a case of balanced X-autosome translocation, schizophrenia and autism in two cases of XX maleness and MLS syndrome (microphthalmia with linear skin defects) in a case of Xp(21.3-pter) deletion. CONCLUSION: Discrepancies between X inactivation ratio and phenotype are not rare and can be due to gene disruption, position effect, complex microrearrangements, variable pattern of X inactivation in different tissues or fortuitous association. In this context, the prognostic value of X inactivation study in prenatal diagnosis will be discussed. PMID- 16690230 TI - Face processing limitation to own species in primates: a comparative study in brown capuchins, Tonkean macaques and humans. AB - Most primates live in social groups which survival and stability depend on individuals' abilities to create strong social relationships with other group members. The existence of those groups requires to identify individuals and to assign to each of them a social status. Individual recognition can be achieved through vocalizations but also through faces. In humans, an efficient system for the processing of own species faces exists. This specialization is achieved through experience with faces of conspecifics during development and leads to the loss of ability to process faces from other primate species. We hypothesize that a similar mechanism exists in social primates. We investigated face processing in one Old World species (genus Macaca) and in one New World species (genus Cebus). Our results show the same advantage for own species face recognition for all tested subjects. This work suggests in all species tested the existence of a common trait inherited from the primate ancestor: an efficient system to identify individual faces of own species only. PMID- 16690231 TI - Development of metagenomic DNA shuffling for the construction of a xenobiotic gene. AB - We describe a metagenomic DNA shuffling process by combining protein engineering process mutation generator and the high potential diversity of metagenomic DNA derived from the environment. Numerous previous shuffling processes attempted to recombine more or less related parental sequences. At the same time, metagenomic approaches unveiled a huge diversity of DNA sequences and genomes, which have not yet been identified to date. In this study, we attempted to combine these two approaches in order to regenerate a novel gene. Here, we present the possibility that DNA fragments from an entire microbial community (metagenome) might be available for the creation of novel genes capable of degrading pollutants. Metagenomic DNA extracted from non-polluted soil was shuffled in vitro to recreate the linA gene responsible for the first steps of lindane degradation. In this work, 74% of the ORF came from separate subsets of the metagenomic pool from a lindane-free and linA-free soil. Our results demonstrate that microbial community genetic diversity can serve as a source for novel gene construction during in vitro manipulation. This in vitro gene construction might also simulate the mosaic nature of novel genes. This demonstration might lead to other attempts to mimic bacterial adaptation and to construct degradative genes for novel compounds not yet released into the environment. PMID- 16690232 TI - Follow-up of twins: health, behaviour, speech, language outcomes and implications for parents. AB - The introduction of assisted reproduction has resulted in a growing number of multiple births. These infants and their parents experience increased, and sometimes unique, medical and psychological risks when compared to singletons. Rates of maternal morbidity, fetal and infant mortality are increased in multiple pregnancies. Twins have a death rate four times higher than singletons and this figure is six times higher for triplets. The main reason is preterm and very preterm birth in multiples, resulting in low and very low birth weight children. Perinatal mortality and morbidity are also more elevated in monozygotic (MZ) twins as compared to dizygotic (DZ) twins. In addition to an increased risk of mortality, multiples have higher rates of morbidity, specifically cerebral palsy and mental subnormality. Language and speech delays are more pronounced in multiples, as are cognitive delays, motor development, behavioural problems and difficulties in parent-child interactions. Depression among parents of multiples is reported to be higher than those of singletons. This paper aims to critically appraise the literature regarding the aforementioned topics, including a comparison between the outcomes for iatrogenic and spontaneously conceived twins and to suggest areas for further research. PMID- 16690233 TI - Growth and development of children conceived by in-vitro maturation of human oocytes. AB - AIMS: The objective of this study is to evaluate the physical and mental development of infants born on in-vitro maturation (IVM) programs. STUDY DESIGN: We compare 21 IVM children as study group and 21 non-IVM children as control group. We performed a general physical examination for malformations, neurological examination, developmental examination (Bayley Scales) on the IVM group and the non-IVM group. RESULTS: We found all the children with normal karyotype and without major malformation in both IVM and non-IVM groups. The mean Mental Development Index scores for IVM subjects and the comparison group were 92.71+/-10.47 and 97.19+/-8.88, respectively (p=0.074). The mean Psychomotor Development Index scores were 96.67+/-8.91 and 96.19+/-7.05, respectively (p=0.817). CONCLUSIONS: This is the first study designed to evaluate the physical growth and developmental indices of IVM children with combinational priming protocol of FSH and hCG. Our results suggest that IVM children didn't show developmental delay during infancy and early childhood. PMID- 16690234 TI - Twin children's language development. AB - Scientific studies have consistently reported that, in group analyses, twin children have higher rates of language delay when compared with single-born children. This paper reviews the evidence on twin language by addressing five key questions:The paper concludes that language delay in twins when compared with singletons is mild yet prevalent, particularly in males and is most likely a result of experience of poorer quality of language environment compared to single born children. Occurrence of "private" or "twin" language is rare and not an explanation for the findings of generalised language delay among twins. Given that language delay in twins relates to social-language experience, appropriate intervention and remediation of language delay should focus on providing a more stimulating language environment both within and outside the home. It is recommended that future research examine, longitudinally, the developmental course and consequences of language delay in twin children and trial interventions designed to improve their language experiences. PMID- 16690235 TI - Antibacterial and antioxidant activities in Sideritis italica (Miller) Greuter et Burdet essential oils. AB - Sideritis italica (Miller) Greuter et Burdet is a widespread Lamiacea in the Mediterranean region used in traditional medicine. Essential oils were antibacterial against nine ATCC and as many clinically isolated Gram-positive and Gram-negative bacterial strains. Antibacterial activity was also found against Helicobacter pylori: a dose-dependant inhibition was shown between 5 and 25 microg/ml. The antibacterial activity of the oils was expressed as MICs (minimum inhibitory concentrations) and MBCs (minimum bactericidal concentrations). At a concentration between 3.9 and 250 microg/ml the oils showed a significant antibacterial effect against both Gram-negative and Gram-positive bacteria. In particular the ATCC strains Pseudomonas aeruginosa (MIC=3.9 microg/ml and 7.8 for flowerheads and leaves, respectively), Proteus mirabilis (MIC=15.6 and 7.8 microg/ml), Salmonella typhi (MIC=7.8 microg/ml) and Proteus vulgaris (MIC=15.6 microg/ml) were the most inhibited. Only Pseudomonas aeruginosa showed MBC at a concentration between 62.6 and 125 microg/ml. The antioxidant activity of the essential oils was evaluated by two cell free colorimetric methods: ABTS and DMPD; leaf oil is more active (4.29 +/- 0.02 trolox equivalents and 4.53 +/- 0.67 ascorbic acid equivalents by ABTS and DMPD, respectively). Finally the antioxidant activity of the essential oils was also evaluated by their effects on human whole blood leukocytes (WB) and on isolated polymorphonucleate (PMN) chemiluminescence. Comparing the effects of the oils from leaves and flowerheads on both PMN and WB chemiluminescence emission, we found no significant differences. Essential oils showed a dose-dependent and linear inhibitory activity on isolated PMN as well as on WB CL emission when PMA-stimulated. On the contrary, the inhibitory activity on resting cells was nonlinear. Our data represent an answer to the continual demand for new antibiotics and antioxidants for the continuous emergence of antibiotic-resistant strains and the growing interest in the substitution of synthetic antioxidants with natural ones. Furthermore, our finding of antibacterial and antioxidant activities in the essential oils from Sideritis italica flowerheads and leaves validated the use of the plant for medical purposes. PMID- 16690236 TI - Fatal Candida tropicalis infection in an 8-month-old infant with an aplasia of the thymus as a rare cause of death in infancy. AB - Anomalies of the thymus can lead to immunodeficiencies that are often associated with complex syndromes. The clinical relevance of those anomalies varies a lot. Regarding mortality and morbidity, opportunistic infections play a major role in immunocompromised individuals. CASE REPORT: An 8-month-old infant died of a sepsis with multi-organ failure after 32 days in intensive care. At autopsy, no thymus gland could be found; toxicological examinations yielded high morphine concentrations. Autopsy findings and the results of further examinations performed postmortem are discussed in view of the clinical findings, the classification and the cause of death. PMID- 16690237 TI - Mitochondrial DNA identification of game and harvested freshwater fish species. AB - The use of DNA in forensics has grown rapidly for human applications along with the concomitant development of bioinformatics and demographic databases to help fully realize the potential of this molecular information. Similar techniques are also used routinely in many wildlife cases, such as species identification in food products, poaching and the illegal trade of endangered species. The use of molecular techniques in forensic cases related to wildlife and the development of associated databases has, however, mainly focused on large mammals with the exception of a few high-profile species. There is a need to develop similar databases for aquatic species for fisheries enforcement, given the large number of exploited and endangered fish species, the intensity of exploitation, and challenges in identifying species and their derived products. We sequenced a 500bp fragment of the mitochondrial cytochrome b gene from representative individuals from 26 harvested fish taxa from Ontario, Canada, focusing on species that support major commercial and recreational fisheries. Ontario provides a unique model system for the development of a fish species database, as the province contains an evolutionarily diverse array of freshwater fish families representing more than one third of all freshwater fish in Canada. Inter- and intraspecific sequence comparisons using phylogenetic analysis and a BLAST search algorithm provided rigorous statistical metrics for species identification. This methodology and these data will aid in fisheries enforcement, providing a tool to easily and accurately identify fish species in enforcement investigations that would have otherwise been difficult or impossible to pursue. PMID- 16690238 TI - MRT-measurements of muscle volumes of the lower extremities of youths with spastic hemiplegia caused by cerebral palsy. AB - After long term of studies from our gait lab, the typical muscular dysbalances by all of our patients with cerebral palsy where pointed out. Now we wanted to examine using the MRT, weather the dysbalances of the hemiparetic musculoskeletal system also show up in discrepancies of the muscle volumes. The MRT slices of the lower extremities were segmented. From this cross sections the muscles volumes were derived. These where analyzed particularly with regard to asymmetries between spastic and healthy side. Hemiparetic patients showed reduced volumes of all muscles on the paretic leg in comparison to the healthy side. The muscles of the thigh of the paretic leg were reduced to 84% in the mean over all muscles and in comparison to the healthy leg. The volume of the muscles of the shank was reduced to 72%, significantly more than the muscles of the thigh. Concerning flexor and extensor muscles located at thigh and shank of the paretic leg there was found significantly the same relative deficit of the muscle volume. Examining the muscle volumes of subjects with different neurogene foot deformities, significant differences of the volume reduction of single muscles appeared within the lower leg. PMID- 16690239 TI - Technical and clinical success of infrarenal endovascular abdominal aortic aneurysm repair: A 10-year single-center experience. AB - OBJECTIVE: The aim of our retrospective study was to review our single-center experience with aortic abdominal aneurysm (AAA) repair retrospectively. MATERIAL AND METHODS: From 1995 to 2005, 70 consecutive patients affected by AAA were treated by endovascular stent-graft repair. Mean follow-up was 23.9 months. Follow-up investigations were performed at 6 and 12 months and yearly thereafter. Five different stent-graft designs were compared to each other. Primary technical success (PTS), assisted primary technical success (APTS), primary clinical success (PCS) and secondary clinical success (SCS) were evaluated. RESULTS: All over PTS was achieved in 94.3%, APTS in 97.1%, PCS in 61.4%, APCS in 64.3% and SCS in 70%. There were 3 type I endoleaks, 25 type II endoleaks, 4 type III endoleaks, 8 limb problems, 5 conversions to open surgery, 10 aneurysm sac expansions and 14 device migrations. Patients with newer generation devices showed better results than patients with first generation prosthesis. In addition results were better for grafts with suprarenal fixation (versus infrarenal fixation) and grafts with barbs and hooks (versus grafts without barbs and hooks). Patients with bad anatomic preconditions showed a higher complication rate. CONCLUSION: Contrary to first generation products, new stent-graft designs show acceptable technical and clinical results in endovascular AAA aneurysm repair. However, this therapy still should be reserved only for patients with significant comorbities and suitable anatomic conditions. PMID- 16690240 TI - Intrahepatic recurrence after percutaneous radiofrequency ablation of hepatocellular carcinoma: analysis of the pattern and risk factors. AB - PURPOSE: To evaluate the pattern and risks for intrahepatic recurrence after percutaneous radiofrequency (RF) ablation for hepatocellular carcinoma (HCC). MATERIALS AND METHODS: We studied 62 patients with 72 HCCs (< or =4 cm) who were treated with percutaneous RF ablation. The mean follow-up period was 19.1 months (6.0-49.1). We assessed the incidence and cumulative disease-free survival of local tumor progression (LTP) and intrahepatic distant recurrence (IDR). To analyze the risk factors, we examined the following, for the LTP: (1) tumor diameter, (2) contact with vessels, (3) degree of approximation to hepatic hilum, (4) contact with hepatic capsule, (5) presence of ablative safety margin, (6) degree of benign periablational enhancement and (7) serum alpha-fetoprotein; for the IDR: (1) severity of hepatic disease, (2) presence of HBsAg, (3) serum alpha fetoprotein, (4) whether RF ablation was the initial treatment and (5) multiplicity of tumor for IDR. RESULTS: The incidence of overall recurrence, LTP and IDR was 62.9%, 26.4% and 53.2%, respectively. The cumulative disease-free survival rates were 52%, 82% and 56% at 1 year, 26%, 63% and 30% at 2 years, respectively. Univariate analysis showed that the significant risk factors for LTP were: a tumor with a diameter >3 cm, contact of HCC with a vessel and an insufficient safety margin (p<0.05). A multivariate stepwise Cox hazard model showed that the measurement of a tumor diameter >3 cm and insufficient safety margin were independent factors. Only the increased serum alpha-fetoprotein was a significant risk factor for IDR (p<0.05). CONCLUSION: Intrahepatic recurrence after percutaneous RF ablation is common. Large HCC (>3 cm) with high serum alpha fetoprotein should be treated more aggressively because of higher risk for recurrence. PMID- 16690241 TI - Outlook for cellulase improvement: screening and selection strategies. AB - Cellulose is the most abundant renewable natural biological resource, and the production of biobased products and bioenergy from less costly renewable lignocellulosic materials is important for the sustainable development of human beings. A reduction in cellulase production cost, an improvement in cellulase performance, and an increase in sugar yields are all vital to reduce the processing costs of biorefineries. Improvements in specific cellulase activities for non-complexed cellulase mixtures can be implemented through cellulase engineering based on rational design or directed evolution for each cellulase component enzyme, as well as on the reconstitution of cellulase components. Here, we review quantitative cellulase activity assays using soluble and insoluble substrates, and focus on their advantages and limitations. Because there are no clear relationships between cellulase activities on soluble substrates and those on insoluble substrates, soluble substrates should not be used to screen or select improved cellulases for processing relevant solid substrates, such as plant cell walls. Cellulase improvement strategies based on directed evolution using screening on soluble substrates have been only moderately successful, and have primarily targeted improvement in thermal tolerance. Heterogeneity of insoluble cellulose, unclear dynamic interactions between insoluble substrate and cellulase components, and the complex competitive and/or synergic relationship among cellulase components limit rational design and/or strategies, depending on activity screening approaches. Herein, we hypothesize that continuous culture using insoluble cellulosic substrates could be a powerful selection tool for enriching beneficial cellulase mutants from the large library displayed on the cell surface. PMID- 16690242 TI - Endovascular abdominal aortic aneurysm repair: long-term outcome measures in patients at high-risk for open surgery. AB - PURPOSE: The study was conducted to determine the outcome in the United States after endovascular repair (EVAR) of infrarenal abdominal aortic aneurysms (AAAs) in patients at high-risk for open surgery by using independently audited, high compliance, chart-verified data sets, and to compare those results with open surgery. METHODS: High-risk was defined to match a recent European trial (EVAR2) and included age of > or =60 years with aneurysm size of > or =5.5 cm, plus at least one cardiac, pulmonary, or renal comorbidity. Data from five multicenter investigational device exemption clinical trials leading to Food and Drug Administration (FDA) approval were analyzed. Of 2216 EVAR patients, 565 met the high-risk criteria. Of 342 surgical controls (OPEN), 61 met high-risk criteria. Primary outcome comparisons included AAA-related death, all-cause death, and aneurysm rupture. Secondary measures were endoleak, AAA sac enlargement, and migration. RESULTS: Average age of the high-risk EVAR subset was 76 +/- 7 years vs 74 +/- 6 years OPEN (P = 0.07), mean EVAR AAA size was 6.4 +/- 0.8 cm vs 6.6 +/- 1.0 cm OPEN (P = .33), and average EVAR follow-up was 2.7 years vs 2.5 years OPEN. The 30-day operative mortality was 2.9% in EVAR vs 5.1% in OPEN (P = .32). The AAA-related death rate after EVAR was 3.0% at 1 year and 4.2% at 4 years compared with 5.1% at both time points after OPEN (P = .58). Overall survival at 4 years after EVAR was 56% vs 66% in OPEN (P = .23). After treatment, EVAR successfully prevented rupture in 99.5% at 1 year and in 97.2% at 4 years. CONCLUSIONS: Endovascular repair of large infrarenal AAAs in anatomically suited high-surgical-risk patients using FDA-approved devices in the United States is safe and provides lasting protection from AAA-related mortality. EVAR mortality remained comparable with OPEN up to 4 years. The decision to treat AAAs in patients with advanced age and significant comorbidities must be individualized and carefully considered, but repair provides excellent protection from AAA related death. PMID- 16690243 TI - Ultra-radical compartmentalized surgery in gynaecological oncology. AB - AIMS: To describe "ultra-radical" surgery in gynaecological oncology. This is multivisceral pelvic resection aimed at the eradication of locally advanced and recurrent gynaecologic cancer with microscopically tumour-free margins (R0). METHODS: Up-dated analysis of a prospective trial evaluating oncologic outcome and treatment-related morbidity of ultra-radical compartmentalized surgery carried out by the author. RESULTS: From 8/1996 until 9/2005 74 patients with locally advanced and recurrent gynaecologic tumours have been treated with ultra radical compartmentalized surgery. Eighteen patients with central disease underwent exenteration as multimesovisceral excision, 56 patients with pelvic side wall disease received laterally extended endopelvic resection. In 72 of the patients the tumours were removed with microscopically free margins (R0). Two patients with advanced age and extensive comorbidity respectively, died during the early postoperative period. Moderate and severe treatment-related morbidity was 66%. At a median follow-up period of 29 months (1-112 months) 5-year overall and recurrence-free survival probabilities are 56% (95% CI: 42-69) and 56% (42 70). CONCLUSION: Ultra-radical compartmentalized pelvic surgery may salvage selected patients with locally advanced and recurrent gynaecologic malignancies including those with pelvic side wall disease traditionally not considered for surgical therapy. PMID- 16690244 TI - Management of vulvar cancers. AB - AIM: The radical surgical approach in the treatment of vulvar cancer patients has led to a favorable prognosis for the majority of the patients with early stage squamous cell cancer. However, the morbidity is impressive, leading to more individualized treatment. The aim of this review is to give an overview of the management of vulvar cancer. METHODS: We have reviewed the literature on the modifications in treatment of vulvar cancer with the emphasis on surgery and radiotherapy for primary disease. RESULTS: While surgery is the cornerstone of treatment for early stage squamous cell vulvar cancer (with wide local excision and uni- or bilateral inguinofemoral lymphadenectomy via separate incisions as standard treatment), until now there has been a limited role for primary radiotherapy although this may be an attractive alternative for the inguinofemoral lymphadenectomy in the future. The sentinel lymph node procedure with the combined technique (preoperative lymphoscintigraphy with (99m)technetium labeled nanocolloid and Patente Blue) is a promising staging technique for patients with vulvar cancer. The clinical implementation of the sentinel lymph node procedure and the role of additional histopathological techniques of the sentinel lymph nodes have to be investigated. In advanced vulvar cancer, chemoradiation followed by surgery should be regarded as the treatment of first choice for these patients. CONCLUSIONS: Due to the rarity of vulvar cancer it is quite clear that further studies will have to be performed by international collaborative groups. The sentinel lymph node procedure and primary radiotherapy are promising methods to reduce morbidity of treatment, but their safety needs to be studied in clinical trials. PMID- 16690245 TI - [Cerebral metabolism and brain injury]. AB - Brain energy metabolism and signal transduction are intimely intricated. At the cellular level this is reflected by the interdependent metabolism of glutamate and glucose and the energetic compartmentalization between astrocytic glycolysis and neuronal metabolism. Astrocytes appear to have a particular importance in brain metabolism by regulating microcirculation and the repartition of energetic substrates in function of synaptic activity. The high level of O(2) consumption compared to the mass of tissue confers a particular vulnerability of brain to oxidative stress. The synthesis of glutathione, the main anti-oxidant of brain, appears to be dependent of the regulation of synaptic glutamate concentration by astrocytes. Deficiencies of astrocytes functions appear to play a key role in the physiopathology of brain injury. PMID- 16690246 TI - [Brain injury during severe sepsis]. AB - Sepsis-associated encephalopathy is a global cerebral dysfunction induced by the systemic response to inflammation and infection, without a liver or renal injury. Alteration of consciousness, from confusion to coma, is the main clinical symptom. This encephalopathy is associated with an increase in mortality due to sepsis. Its physiopathology is unknown. There is frequently an increased permeability of the blood-brain barrier, which might explain a role of endotoxins on cerebral metabolism. Changes in neurotransmitter release or concentrations (norepinephrine, serotonin, dopamine, GABA) have been reported. There is not any specific treatment of septic encephalopathy. In most cases, this syndrome is rapidly reversible after the treatment of sepsis. PMID- 16690247 TI - [Cerebral ischemic threshold in clinical practice]. AB - The ischemic threshold is reached when the availability of oxygen in the cerebral tissue does not cover oxygen requirement. For a patient sedated, with constant PaO(2) and haemoglobin, the cerebral blood flow (CBF) global and local is the essential factor to maintain such a balance. At a cellular level, ischemia occurs when the CBF is below 20-25 ml/min. However, this threshold probably varies with the patient and also within the normal or perilesional tissue. A cerebral perfusion pressure (CPP) of 60 mmHg, recommended for a cerebral perfusion allowing a sufficient CBF for normal brain, does not prevent ischemia. Monitoring aimed to control parameters of the aerobic metabolism (PtiO(2), SjO(2) and microdialysis) and to detect the ischemic threshold allows to adapt the CPP to each patient and continuously. PMID- 16690248 TI - Tandem repeats in the CpG islands of imprinted genes. AB - In contrast to most genes in mammalian genomes, imprinted genes are monoallelically expressed depending on the parental origin of the alleles. Imprinted gene expression is regulated by distinct DNA elements that exhibit allele-specific epigenetic modifications, such as DNA methylation. These so called differentially methylated regions frequently overlap with CpG islands. Thus, CpG islands of imprinted genes may contain special DNA elements that distinguish them from CpG islands of biallelically expressed genes. Here, we present a detailed study of CpG islands of imprinted genes in mouse and in human. Our study shows that imprinted genes more frequently contain tandem repeat arrays in their CpG islands than randomly selected genes in both species. In addition, mouse imprinted genes more frequently possess intragenic CpG islands that may serve as promoters of allele-specific antisense transcripts. This feature is much less pronounced in human, indicating an interspecies variability in the evolution of imprinting control elements. PMID- 16690249 TI - Synthetic, pure-phase beta-tricalcium phosphate ceramic granules (Cerasorb) for bone regeneration in the reconstructive surgery of the jaws. AB - The aim of this study was to investigate the long-term effect of the ceramic beta tricalcium phosphate (beta-TCP) at different sites of alveolar reconstruction and to evaluate its properties. From 1997 to 2002, beta-TCP was implanted as bone substitute in 152 patients using a standardized study protocol. Main indications were the filling of large mandibular cysts (n=52), secondary and tertiary alveolar cleft grafting (n=38), periodontal defects (n=24) and maxillary sinus floor augmentation (n=16). For defects exceeding 2cm in diameter, beta-TCP was combined with autologous bone taken from the retromolar area, the maxillary tuberosity or the chin region. A radiological, clinical and ultrasonographical examination was carried out 4, 12 and 52 weeks postoperative. In 16 cases, biopsies were taken after 12 months indicating complete bony regeneration. While wound-healing disturbances occurred in 9.2% of cases, partial loss of the bone substitute material was found in 5.9%, while total loss occurred in only 2%. Complete radiological replacement of beta-TCP by autologous bone was found after approximately 12 months, indicating its osteoconductive properties. Because of its versatility, low complication rate and good long-term results, synthetic, pure-phase beta-TCP is a suitable material for the filling of bone defects in the alveolar region. PMID- 16690250 TI - A new approach to repairing cleft palate and acquired palatal defects with distraction osteogenesis. AB - Cleft palate (CP) is one of the most common human congenital deformities, and acquired palate defects after trauma or tumour resection are also common. In this study, distraction osteogenesis (DO) for CP and other palatal bone defects was evaluated. Twenty cats were assigned randomly to 3 groups of (1) 15, (2) 3 and (3) 2 cats. In groups 1 and 2, a rectangular ostectomy, in the posterior of the palatal bone shelf, was performed in the sagittal axis to establish the CP defect model. At the same time, a pure titanium intraoral distractor was fixed to molar teeth with brackets and to the palatal bone shelf across the defect with titanium miniscrews bilaterally. Four weeks later, a secondary transport disc (TD) osteotomy was performed, and gradual DO treatment started at 0.4mm twice a day, after 6 days of latency. DO was performed until the TD reached the opposite margin over the gap in 5-6 days. Three cats each of group 1 were killed at 2, 4, 6, 8 and 12 weeks after completion of DO. In group 2, the bone and soft-tissue defects were untreated until death 6 weeks later. Group 3 cats (control) were killed after 6 weeks. The TD successfully recombined with the opposite palatal bone stump, and proportional expansion of the overlay mucoperiosteal flap was achieved. Intramembranous bone formation was revealed: parallel collagen bundles gradually deposited on new bone trabeculae while the proliferative osteoblasts produced bone matrix. The bone defect was finally reconstructed by de novo osteogenesis. The control group was observed to have no spontaneous repairing. These results suggest that the CP defect was reconstructed by osteogenesis in situ, and the soft tissues expanded simultaneously to achieve functional correction. The intraoral distractor provided both effective distraction and stability. PMID- 16690251 TI - A new navigation system based on cephalograms and dental casts for oral and maxillofacial surgery. AB - Intraoperative navigation systems help surgeons to accurately carry out preoperative plans without injuring anatomically important structures. A system is evaluated that uses cephalograms instead of computed tomographic (CT) scans to create images. Three-dimensional (3D) dental casts provide registration between imaging data and the patient. Cephalograms are widely employed in orthognathic and oral and maxillofacial surgery and expose patients to lower doses of radiation than CT. The system uses a dental cast to register the operation field to a pair of frontal and lateral cephalograms. The cast is transformed to 3D data with a laser scanner and a programme that runs on a personal computer. 3D data describing the dental cast, cephalograms and the oral and maxillofacial region of the patient are integrated with specialized software. The optical tracking system for navigation uses charged-coupled-device (CCD) video cameras and light-emitting diodes (LEDs). Two CCD video cameras follow the 3D coordinates of LED assemblies attached to the head, lower jaw and a handpiece. Errors occurring when a dental cast was transformed to 3D data ranged from 0.08 to 0.21 mm. Mean errors were 0.71 mm (0.21-1.09 mm) for the right maxillary central incisor, 0.62 mm (0.04 1.69 mm) for the right maxillary 2nd molar and 1.02 mm (0.23-1.47 mm) for the left maxillary 2nd molar. This surgical navigation system is sufficiently accurate for use in oral and maxillofacial surgery. PMID- 16690252 TI - Differential expression of apoptosis-related proteins in various cellular components of ameloblastomas. AB - To evaluate the expression patterns of apoptosis-related proteins, including Fas, Fas-ligand (FasL), caspase-3 and Bcl-2, in various cellular components of ameloblastomas, 39 cases of ameloblastoma were examined using immunohistochemistry. The staining intensity of the antigens in the 4 types of tumour cellular component, peripheral basal cells of tumour nests, central stellate reticulum-like cells, and foci of squamous and granular cells, was scored using a semi-quantitative scale, and comparisons were made by statistical analysis. Expression of Fas, FasL and caspase-3 was detected in the majority of cases, with a similar pattern of strong staining in the foci of squamous metaplasia and granular cells usually situated in the central area of tumour islands. In contrast, expression of Bcl-2 was predominantly seen in the peripheral basal cell layer. There were significant differences in the staining intensity of Fas, caspase-3 and Bcl-2 among the 4 types of tumour cell. The differential expression of apoptosis-related proteins in various cellular components of ameloblastomas, with pro-apoptotic proteins, Fas, FasL and caspase 3 being closely associated with squamous metaplasia and granular transformation of the tumour cells, suggests that Fas/FasL-induced apoptotic cell death may play a role in the disposal of terminally differentiated or degenerative tumour cells in ameloblastomas. PMID- 16690253 TI - The application of mini-implants for orthodontic anchorage. AB - The aim of this study was to explore the use of mini-implants for skeletal anchorage, and to assess their stability and the causes of failure. Forty-five mini-implants were used in orthodontic treatment. The diameter of the implants was 2mm, and their lengths were 8, 10, 12 and 14mm. The drill procedure was directly through the cortical bone without any incision or flap operation. Two weeks later, a force of 100-200g was applied by an elastometric chain or NiTi coil spring. Risk factors for the failure of mini-implants were examined statistically using the Chi-square or Fisher exact test as applicable. The average placement time of a mini-implant was about 10-15min. Four mini-implants loosened after orthodontic force loading. The overall success rate was 91.1%. The location of the implant was the significant factor related to failure. In conclusion, the mini-implants are easy to insert for skeletal anchorage and could be successful in the control of tooth movement. PMID- 16690254 TI - Associating functional recovery with neurocognitive profiles identified using partially ordered classification models. AB - Neurocognitive deficits are a core feature of schizophrenia and a significant cause of functional disability. However, targeting these deficits with new treatment approaches will only yield functional improvements if those cognitive operations that are responsible for different dimensions of functional recovery can be identified. A major challenge is that conventional neuropsychological tests, the most practical tools for broadly sampling cognitive functions in treatment trials, are polyfactorial, so that task performance is influenced by multiple cognitive operations. Hence, it is difficult to pinpoint exactly for which cognitive operations a low scoring subject may have poor functionality. We have previously applied in a neuropsychological test battery administered to 220 patients having schizophrenia or schizoaffective disorder, Bayesian statistical methods (yielding partially ordered sets, or posets) designed to mimic the expert analysis of a neuropsychologist by classifying patients into discrete groupings or "states" each having a unique cognitive profile. Here, we report on the association of attributes describing these states (viz. working memory, capacity for divergent thinking, cognitive flexibility and psychomotor speed) with two domains of functional outcome (work/education and residential functioning) rated up to 18 months later. After multiplicity correction, only working memory was associated with work/education outcome. While working memory was not associated with residential outcome, the remaining three attributes were. These findings suggest that different neurocognitive operations may be responsible for different outcome domains. Findings support the use of the poset methodology for clarifying patterns of relationships between discrete neurocognitive attributes and domains of functional outcome. PMID- 16690255 TI - Deficits in memory strategy use are related to verbal memory impairments in adolescents with schizophrenia-spectrum disorders. AB - OBJECTIVE: To assess the nature of learning and verbal memory deficits in adolescents with schizophrenia-spectrum disorders (SzS) (i.e., schizophrenia, schizoaffective disorder, and schizophreniform disorder). METHOD: Sixty patients with SzS (mean age=16.1 years, S.D. = 2.2) and 60 age- and gender-matched diagnosis-free healthy volunteers were assessed using the California Verbal Learning Test (CVLT). Planned analyses were conducted to assess the following aspects of memory: span of apprehension, verbal learning, short-term and long term memory, rate of forgetting, interference, and organizational strategies. Adolescents with schizophrenia (Sz) were compared to those with schizoaffective disorder (SzA). Second, patients' test profiles were compared to those of controls. Relationships between initial learning and overall verbal learning with organizational strategy were explored. RESULTS: Neurocognitive profiles did not significantly differ between Sz and SzA participants. Patients performed significantly worse than healthy comparison subjects on measures of span of apprehension, verbal learning, short- and long-term memory, and organizational strategies after adjusting for differences in premorbid intelligence. No group differences were found in rate of forgetting or susceptibility to proactive or retroactive interference. CONCLUSIONS: Adolescents with SzS are characterized by significant verbal memory dysfunction similar to what has been observed in adults with first-episode schizophrenia. Deficits in consistency of learning over several trials, as well as a strong relationship between semantic organizational strategies and reduced learning capacity, implicate dysfunction of the dorsolateral prefrontal cortex as a contributor to verbal memory deficits in adolescents with SzS. PMID- 16690256 TI - Neurocognition and suicidal behaviour in an Irish population with major psychotic disorders. AB - OBJECTIVES: Although neurocognitive deficits are seen as core to schizophrenia the association between suicidality and neurocognition has received little attention. Our aim was to examine the relationship between neurocognitive variables and suicidal behaviour in patients with schizophrenia and schizoaffective disorder. METHODS: Seventy-eight patients with DSM-IV diagnoses of schizophrenia or schizoaffective disorder were categorised as either having attempted suicide or not having attempted suicide based on clinical interview and chart review. Attempters and non-attempters were compared on an extensive neuropsychological battery examining pre-morbid and current general cognitive functioning, episodic memory, and executive functioning. RESULTS: Suicide attempters tended to out perform non-attempters across all areas of executive functioning, and showed significantly better performances on measures of attention and verbal fluency. After controlling for relevant clinical and demographic variables, the differences between attempters and non-attempters remained significant for measures of attention (F = 4.97, p = 0.03) and verbal fluency (F = 4.28, p = 0.04). CONCLUSION: This study adds to existing data that suicide attempters with schizophrenia or schizoaffective disorder may have higher cognitive functioning than non-attempters. In particular, the preservation of higher executive function may influence the ability to initiate and plan suicidal behaviour. PMID- 16690257 TI - Usefulness of glycosylated haemoglobin (HbA1c) to screen for diabetes in patients with schizophrenia. PMID- 16690259 TI - First report of Morganella morganii producing CTX-M-15 beta-lactamase. PMID- 16690258 TI - Phylogenetic diversity of culturable bacteria from alpine permafrost in the Tianshan Mountains, northwestern China. AB - Microbes have been discovered in permafrost sediments for nearly a century. However, microbiological analyses of alpine permafrost are very scarce. This study is a first attempt to describe the phylogenetic diversity of a culturable bacterial community isolated from alpine permafrost in the Tianshan Mountains in northwestern China. Aerobic 2.5-6.0x10(5) CFU/gdw (CFU per 1 gram of dry weight) on modified PYGV medium were recovered from alpine permafrost samples at 4 degrees C; among these, 91 bacterial isolates with different morphotypes were characterized by phenotypic properties, such as morphology, colony pigmentation, Gram staining, endospore formation and temperature range of growth. The isolates were further categorized based on amplified rDNA restriction analysis (ARDRA), and 51 representative isolates possessing distinct ARDRA patterns selected for subsequent 16S rDNA sequencing and phylogenetic analysis. The phylogenetic trees placed the 51 isolates in four major groups: the high-G+C Gram-positives, the low G+C Gram-positives, Proteobacteria and the Cytophaga-Flavobacterium-Bacteroides (CFB) phylum. The most abundant and diverse isolates were members of Gram positive bacteria, particularly the Arthrobacter as a dominant group in alpine permafrost culturable populations. Results of the Jukes-Cantor evolutionary distance matrix suggested that the vast majority of the isolates were different strains of known species, and three may represent new species within the genus Chryseobacterium of the CFB phylum. From this study, it is proposed that alpine permafrost sediments in the Tianshan Mountains provide a specific ecological niche for prolonging survival of diverse microbial lineages. PMID- 16690260 TI - Integron presence in a multiresistant Morganella morganii isolate. AB - A multiresistant strain of Morganella morganii was isolated from a patient affected by several severe pathologies. The isolate was found to be resistant to the following antimicrobials: ampicillin, nalidixic acid, cefalothin, cefoxitin, ceftriaxone, ciprofloxacin, chloramphenicol, streptomycin, erythromycin, gentamicin, novobiocin, penicillin, rifampicin, tetracycline and violet crystal. Mechanisms leading to this multiresistance were studied. Porins of M. morganii multiresistant and wild-type strains were analysed by sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) and were characterised by their ability to form channels in planar black lipid bilayers. The channels formed by porins from multiresistant and susceptible strains suggested that the porins of the multiresistant strain were not responsible for resistance. A 6.6 kb plasmid (pML2003) was detected, isolated and studied. pML2003 included two integrons. Direct sequencing revealed that one of the integrons contained two cassettes, aminoglycoside adenyltransferase (aadB) and chloramphenicol acetyltransferase (catB3) conferring resistance to aminoglycosides and chloramphenicol, respectively. The second integron contained carbenicillinase (blaP1b) and adenyltransferase (aadA2), which confer resistance to beta-lactamases and streptomycin, respectively. PMID- 16690261 TI - Evaluation of MRSA Select, a new chromogenic medium for the detection of nasal carriage of methicillin-resistant Staphylococcus aureus. AB - MRSA Select is a new chromogenic medium for the detection from clinical specimens of methicillin-resistant Staphylococcus aureus (MRSA) growing as pink colonies. Screening samples (n=666) taken from patients were spread onto MRSA Select, oxacillin resistance screening agar base (ORSAB), mannitol salt agar and an in house medium containing tobramycin-colistin-fungizone (TCF). Ninety-nine samples were positive for MRSA on at least one selective medium. Sensitivity and specificity were >94% for all media provided that incubation was extended to 48 h for ORSAB and that a coagulase test was performed for ORSAB and TCF. By contrast, on the single criterion of the colour of colonies for MRSA positivity, the specificity and sensitivity of MRSA Select were high (99.8% and 99%, respectively). PMID- 16690262 TI - Life events exposure before a treated major depressive episode depends on the number of past episodes. AB - A cross-sectional survey investigated the relationship between the number of previous depressive episodes and life events, testing the kindling hypothesis, in a sample of 13,377 treated patients with unipolar depression. A linear decline of average life events exposure is observed for more frequent past episodes, even when age, gender and severity are taken into account. PMID- 16690263 TI - Effect of the composition of lecithin/n-propanol/isopropyl myristate/water microemulsions on barrier properties of mice skin for transdermal permeation of tetracaine hydrochloride: in vitro. AB - Effect of composition of lecithin water-in-oil and oil-in-water microemulsion on in vitro transdermal permeation of tetracaine hydrochloride was studied on mice model. The results were compared with an aqueous solution of tetracaine hydrochloride (2.7 mg/ml). In vitro skin flux and permeability coefficients were obtained using the Franz diffusion cell. Differential scanning calorimetry (DSC), transmission electron microscopy (TEM) and confocal laser scanning microscopy (CLSM) were used to study the mechanism of action of the microemulsion. Micrographs of TEM and CLSM studies were analyzed by using Image Pro Plus image software. Skin flux of tetracaine hydrochloride was found to be dependent on the composition of lecithin/n-propanol/isopropyl myristate/water microemulsions. At lower Km ratio (i.e. 0.5:1 and 0.8:1) of microemulsion, the rate of permeation of tetracaine hydrochloride was higher when compared to the microemulsion of higher Km ratio (1:1 and 1.5:1). Image analysis of TEM micrograph, 6h after application of lecithin microemulsion, showed 3.5+/-0.75-fold (p<0.001) increase in the intercellular space in the epidermis and 3.8+/-0.4-fold (p<0.001) enhancement in upper dermis. CLMS results show that sweat gland and hair follicles also provided path for permeation of the drug through the skin. PMID- 16690264 TI - [Child end of life: are guidelines useful?]. PMID- 16690265 TI - [Misleading presentations of celiac disease]. PMID- 16690266 TI - [Preoperative evaluation of cardiac function in children]. PMID- 16690267 TI - [Overuse injuries in the young athletes]. PMID- 16690268 TI - [Will serologic tests allow to diagnose coeliac disease in children?]. PMID- 16690269 TI - [A median lumbar cutaneous fistula in a 3-year-old child]. PMID- 16690270 TI - [Childhood cancer surviving]. PMID- 16690272 TI - [Deceptive symptoms and analysis of diagnostic errors in pediatric emergencies]. PMID- 16690271 TI - [Epidemiology of autoimmune haemolytic anemia in children: French data]. PMID- 16690273 TI - [New treatments for systemic diseases]. PMID- 16690274 TI - [Neurologically impaired children and digestive problems]. PMID- 16690275 TI - [Relationships between children and dogs, prevention of bites]. PMID- 16690276 TI - [Assessing asthma control?]. PMID- 16690277 TI - [Vaccines' adverse events: questions and answers brought by InfoVac]. PMID- 16690278 TI - [Can clinical epidemiology in emergency care improve acute gastroenteritis management in infants?]. PMID- 16690279 TI - [Value of regional anaesthesia for lower limbs surgery in paediatrics]. PMID- 16690280 TI - [Conditioning of children (and their parents) to imaging examination]. PMID- 16690281 TI - [Infant asthma: sole role of respiratory syncytial virus?]. PMID- 16690282 TI - [Bone mineral content measurement in children and adolescents. Analysis and results]. PMID- 16690283 TI - [Fibromyalgia]. PMID- 16690284 TI - [Head injury in children less than 2 years of age]. PMID- 16690285 TI - [Needs' assessment and resources for pediatric palliative care]. PMID- 16690286 TI - [Relationship between ENT and digestive disorders]. PMID- 16690287 TI - [Dental facial orthopedics in young children]. PMID- 16690288 TI - [Organisation of health care management for children with traumatic brain or spinal injuries]. PMID- 16690289 TI - [The use of soy in infant and child feeding]. PMID- 16690290 TI - [Dog bites in children, from epidemiology to management]. PMID- 16690291 TI - [Pediatric infections network]. PMID- 16690292 TI - [Infective endocarditis in children]. PMID- 16690293 TI - [Nose and asthma in children]. PMID- 16690294 TI - [Surgical management of cerebral palsy]. PMID- 16690295 TI - [Chronic autoimmune hemolytic anemia: toward new treatments?]. PMID- 16690296 TI - [Long-term outcomes of urinary flow impairement on renal hemodynamics: from animal experiments to clinical research]. PMID- 16690297 TI - [Epidemiology and prevention of head trauma in children]. PMID- 16690298 TI - [Management of acute phase of autoimmune haemolytic anemia]. PMID- 16690299 TI - [Epidemiology in the pediatric emergency department: needed for children or pediatricians?]. PMID- 16690300 TI - [The French association for Evans syndrome and autoimmune hemolytic anemia]. PMID- 16690301 TI - [Non invasive ventilation in children]. PMID- 16690302 TI - [How to avoid an inaccurate infant formula]. PMID- 16690303 TI - [Soy and phytoestrogens]. PMID- 16690304 TI - [Epidemiology of bacterial meningitis in children in France]. PMID- 16690305 TI - [Ciliary dyskinesia, bronchopulmonary manifestations]. PMID- 16690306 TI - [Movement disorders in children]. PMID- 16690307 TI - [Musculoskeletal pain: diagnostic possibilities]. PMID- 16690308 TI - [Pathophysiology of retinopathy of prematurity]. PMID- 16690309 TI - Knowledge-based data analysis and interpretation. PMID- 16690310 TI - Concentration dependent atrazine-atrazine complex formation promotes selectivity in atrazine imprinted polymers. AB - An atrazine (ATR) molecularly imprinted polymer (MIP) was prepared using a non covalent strategy. The affinity and selectivity of the polymer was initially evaluated under non-equilibrium conditions and the polymer was shown to possess good template selectivity. The selectivity of the polymer was further investigated under equilibrium conditions and over a range of concentrations using Scatchard plots and Hill plots and by assessing distribution coefficients and normalised selectivity values. It was observed that both selectivity and affinity were dependent on the concentration of the ligand and that unusually selectivity and affinity were better at higher atrazine concentrations. It was concluded that this phenomenon resulted from the formation of atrazine-atrazine complexes during the pre-polymerisation stage and during rebinding and that the polymer demonstrated improved atrazine affinity when the conditions favoured complex formation. PMID- 16690311 TI - Importin-mediated nuclear transport in neurons. AB - The polarized morphology of neurons poses a particular challenge to intracellular signal transduction. Local signals generated at distal sites must be retrogradely transported to the nucleus to produce persistent changes in neuronal function. Such communication of signals between distal neuronal compartments and the nucleus occurs during axon guidance, synapse formation, synaptic plasticity and following neuronal injury. Recent studies have begun to delineate a role for the active nuclear import pathway in transporting signals from axons and dendrites to the nucleus. In this pathway, soluble cargo proteins are recognized by nuclear transport carriers, called importins, which mediate their translocation from the cytoplasm into the nucleus. In neurons, importins might serve an additional function by carrying signals from distal sites to the soma. PMID- 16690312 TI - Synthesis and structure-activity relationships of piperidine-based melanin concentrating hormone receptor 1 antagonists. AB - Isosteric replacement of the urea group of lead compound 1 led to novel substituted piperidine phenylamide analogues. SAR on the electron-induced effects of various linkers as well as substituents on the phenyl rings and the piperidine nitrogen has been investigated. Many single-digit nanomolar MCH R1 antagonists have been identified from this series. PMID- 16690313 TI - Inactivation of GABA transaminase by 4-acryloylphenol. AB - Previous study showed that 4-hydroxybenzaldehyde is a competitive inhibitor of GABA transaminase. As a result, 4-acryloylphenol was synthesized as a 4 hydroxybenzaldehyde analogue, and shown to inactivate potently the enzyme in a time-dependent manner. The inactivation was protected by alpha-ketoglutarate, indicating that it occurs at the active site of the enzyme. Beta-mercaptoethanol also prevented the enzyme from inactivation. The possible mechanism involving a Michael addition was proposed to rationalize the inactivation. PMID- 16690314 TI - BACE-1 inhibition by a series of psi[CH2NH] reduced amide isosteres. AB - A series of beta-site amyloid precursor protein cleaving enzyme (BACE-1) inhibitors containing a psi(CH2NH) reduced amide bond were synthesized. Incorporation of this reduced amide isostere as a non-cleavable peptide surrogate afforded inhibitors possessing low nanomolar potencies in both an enzymatic and cell-based assay. PMID- 16690315 TI - Design and synthesis of orally efficacious benzimidazoles as melanin concentrating hormone receptor 1 antagonists. AB - Biaryl urea lead compound 1 was discovered earlier in our MCH antagonist program. Novel benzimidazole analogues with increased chemical stability, devoid of the potential carcinogenic liability associated with a biarylamine moiety, were synthesized and evaluated to be potent MCH R1 antagonists. Two compounds in this series have demonstrated in vivo efficacy in a rodent obesity model. PMID- 16690316 TI - Demyelinating polyneuropathy in Leber hereditary optic neuropathy. AB - We report a patient with Leber hereditary optic neuropathy (G11778A mtDNA) and a severe demyelinating neuropathy, for which no other cause except his mitochondrial disorder could be found. The involvement of the peripheral nervous system of patients with LHON, in particular with a 11778 mtDNA, is discussed. PMID- 16690317 TI - The where's and when's of kinase anchoring. AB - Kinase anchoring has gained acceptance as a means to synchronize spatial and temporal aspects of cell signaling. A-kinase anchoring proteins (AKAPs) are a diverse group of functionally related proteins that target protein kinase A and other enzymes to coordinate a range of signaling events. Recent advances in this field have shown that incorporating phosphodiesterases into AKAP signaling complexes exerts local control of cAMP metabolism, that phosphorylation of some AKAPs potentiates downstream signaling events, that anchoring of distinct enzyme combinations functions as a mechanism to expand the repertoire of cellular events controlled by a single AKAP, and that fluorescent biosensors can be used to visualize dynamic aspects of localized cAMP signaling. PMID- 16690318 TI - New analogues of butylated hydroxytoluene as anti-inflammatory and antioxidant agents. AB - Amine or amide derivatives bearing the 2,6-di-tert-butyl phenol moiety are synthesised. Almost all are antioxidants, reduce acute inflammation and inhibit COX-1 and lipoxygenase activity. The most potent anti-inflammatory, COX-1 inhibitor and antioxidant agent, with low toxicity, is 2,6-di-tert-butyl-4 thiomorpholin-4-ylmethyl-phenol. PMID- 16690319 TI - Application of multicomponent reactions to antimalarial drug discovery. Part 2: New antiplasmodial and antitrypanosomal 4-aminoquinoline gamma- and delta-lactams via a 'catch and release' protocol. AB - A parallel synthesis of a new series of 4-aminoquinoline gamma- and delta-lactams synthesized via the Ugi 3-component 4-centre multicomponent reaction is described. The basicity of the quinoline nitrogen was exploited in the purification of compounds via a 'catch and release' protocol. Yields ranging from 60% to 77% and purities as high as 96% were obtained. Compound 29, the most active against a chloroquine-resistant W2 strain of Plasmodium falciparum with an IC(50) of 0.096 microM, also inhibited recombinant falcipain-2 in vitro (IC(50)= 17.6 microM). Compound 17 inhibited the growth of Trypanosoma brucei with an ED(50) of 1.44 microM whilst exhibiting a favourable therapeutic index of 409 against a human KB cell line. PMID- 16690320 TI - Role of endothelial nitric oxide synthase in cerebral blood flow changes during kainate seizures: a genetic approach using knockout mice. AB - The role of endothelial nitric oxide (NO) in the cerebrovascular response to partial seizures was investigated in mice deleted for the endothelial NO synthase gene (eNOS-/-) and in their paired wild-type (WT) congeners. Local cerebral blood flow (LCBF, quantitative [14C]iodoantipyrine method) was measured 3-6 h after unilateral kainate (KA) injection in the dorsal hippocampus; controls received saline. In WT mice, KA seizures induced a 22 to 50% LCBF increase restricted to the ipsilateral hippocampus, while significant LCBF decreases (15-33%) were noticed in 22% of the contralateral areas, i.e., the parietal cortex, amygdala and three basal ganglia areas, compared to saline-injected WT mice. In eNOS-/- mice, no LCBF increases were recorded within the epileptic focus and generalized contralateral LCBF decreases (22-46%) were noticed in 2/3 of the brain areas, compared to saline-injected eNOS-/- mice. Thus, endothelial NO is the mediator of the cerebrovascular response within the epileptic focus and participates in the maintenance of LCBF in distant areas. PMID- 16690321 TI - p53 as a target for anti-cancer drug development. AB - Loss of p53 function compromises genetic homeostasis in cells exhibiting deregulated DNA replication and/or DNA damage, and prevents normal cytotoxic responses to cancer therapies. Genetic and pharmacological approaches are being developed with the ultimate goal of restoring or controlling p53 functions in cancer patients. Progress has recently been made in the clinical use of replication-deficient virus carrying wt-TP53 (Ad5CMV-p53) and/or cancer-selective oncolytic adenoviruses (ONYX-015). These strategies demonstrated clinical activity as monotherapy and were synergistic with traditional chemotherapy agents in the treatment of some types of cancer. In addition, pharmacological methods are under development to either stimulate wild-type p53 protein function, or induce p53 mutant proteins to resume wild-type functions. These methods are based on small chemicals (CP-31388, PRIMA-1), peptides (CDB3) or single-chain Fv antibody fragments corresponding to defined p53 domains. Here, we discuss the mechanisms underlying these approaches and their perspectives for cancer therapy. PMID- 16690322 TI - Functional and structural characterization of the Methanosarcina mazei proteasome and PAN complexes. AB - We have cloned the proteasome and the proteasome activating nucleotidase (PAN) genes from the mesophilic archaeon Methanosarcina mazei and produced the respective proteins in Escherichia coli cultures. The recombinant complexes were purified to homogeneity and characterized biochemically, structurally, and by mass spectrometry. We found that the degradation of Bodipy-casein by Methanosarcina proteasomes was activated by Methanosarcina PAN. Notably, the Methanosarcina PAN unfolded GFP-SsrA only in the presence of Methanosarcina proteasomes. Structural analysis by 2D averaging electron microscopy of negatively stained complexes displayed the typical structure for the proteasome, namely four-striped side-views and sevenfold-symmetric top-views, with 15 nm height and 11 nm diameter. The structural analysis of the PAN preparation revealed also four-striped side-views, albeit with a height of 18 nm and sixfold symmetric top-views with a diameter of 15 nm, which corresponds most likely to a dimer of two hexameric complexes. Mass spectrometric analysis of both the Methanosarcina and the Methanocaldococcus PAN proteins indicated hexameric complexes. In summary, we performed a functional and structural characterization of the PAN and proteasome complexes from the archaeon M. mazei and described unique new structural and functional features. PMID- 16690323 TI - Efficient implementation of a filtered back-projection algorithm using a voxel-by voxel approach. AB - The large amount of image data necessary for high-resolution 3D reconstruction of macromolecular assemblies leads to significant increases in the computational time. One of the most time consuming operations is 3D density map reconstruction, and software optimization can greatly reduce the time required for any given structural study. The majority of algorithms proposed for improving the computational effectiveness of a 3D reconstruction are based on a ray-by-ray projection of each image into the reconstructed volume. In this paper, we propose a novel fast implementation of the "filtered back-projection" algorithm based on a voxel-by-voxel principle. Our version of this implementation has been exhaustively tested using both model and real data. We compared 3D reconstructions obtained by the new approach with results obtained by the filtered Back-Projections algorithm and the Fourier-Bessel algorithm commonly used for reconstructing icosahedral viruses. These computational experiments demonstrate the robustness, reliability, and efficiency of this approach. PMID- 16690324 TI - Expression and phylogenetic analysis of the ninth complement component (C9) in rainbow trout. PMID- 16690325 TI - Cloning and characterisation of two natural killer enhancing factor genes (NKEF-A and NKEF-B) in pufferfish, Tetraodon nigroviridis. AB - Natural killer cell enhancing factor (NKEF) belongs to a highly conserved peroxiredoxin (Prx) family. Its functions are involved in immune cytotoxicity, apoptosis, cell proliferation, differentiation and antioxidant activity. In present paper, two NKEF genes, NKEF-A and NKEF-B, were isolated and characterised in spotted green pufferfish, Tetraodon nigroviridis. Comparative genomic location analysis shows conserved syntenies within the NKEF-A and NKEF-B regions between Tetraodon and human, indicating that the Tetraodon NKEF-A and NKEF-B genes are orthologs for mammalian NKEF-A and NKEF-B encoding genes, respectively. The genomic organisation of the two genes is similar in different species, containing one non-coding exon, five coding exons and five introns. The deduced amino acids of the two genes are highly homologous with mammalian NKEF-A and NKEF-B proteins, including two well conserved Val-Cys-Pro (VCP) motifs. Tissue specific expression analysis by RT-PCR reveals that both genes are globally expressed. Stimulation with LPS inhibits both NKEF-A and NKEF-B gene expression in most tissues but enhances NKEF-A and NKEF-B transcriptions in spleen. This is the first report on the existence of a NKEF-B homologue in fish. PMID- 16690326 TI - Muscle enhancement using closed-loop electrical stimulation: volitional versus induced torque. AB - In cases of partial deficiency of muscle activation capacity, force augmentation can be achieved by hybrid activation, i.e., by combining electrical stimulation (ES) with volitional activation. In this activation modality the shares of the volitional and induced torques within the overall hybrid torque are unknown. The purpose of this study was to suggest a computational approach to parcel out the volitional and stimulation induced components of joint torque generated during combined voluntary and electrical activation of the Tibialis Anterior muscle (TA). For this purpose, isometric contraction of the TA was studied on 5 healthy subjects, using an activation protocol involving ES alone, volitional activation alone and hybrid activation. Ankle torque and TA EMG were measured. A computational algorithm was developed to dissociate the volitional from the overall torque, based on EMG filtering and on pre-measured calibration curves of volitional torque versus EMG. The results indicated that for a certain hybrid torque there is a linear decaying relationship between the induced torque and the volitional torque shares. Moreover, based on a defined enhancement ratio, the results indicate that within the range of stimulation intensities, there exist regions of increased facilitation, in which the stimulation efficiency is higher under combined compared to isolated conditions. PMID- 16690327 TI - A modern approach to rotiferan phylogeny: combining morphological and molecular data. AB - The phylogeny of selected members of the phylum Rotifera is examined based on analyses under parsimony direct optimization and Bayesian inference of phylogeny. Species of the higher metazoan lineages Acanthocephala, Micrognathozoa, Cycliophora, and potential outgroups are included to test rotiferan monophyly. The data include 74 morphological characters combined with DNA sequence data from four molecular loci, including the nuclear 18S rRNA, 28S rRNA, histone H3, and the mitochondrial cytochrome c oxidase subunit I. The combined molecular and total evidence analyses support the inclusion of Acanthocephala as a rotiferan ingroup, but do not support the inclusion of Micrognathozoa and Cycliophora. Within Rotifera, the monophyletic Monogononta is sister group to a clade consisting of Acanthocephala, Seisonidea, and Bdelloidea-for which we propose the name Hemirotifera. We also formally propose the inclusion of Acanthocephala within Rotifera, but maintaining the name Rotifera for the new expanded phylum. Within Monogononta, Gnesiotrocha and Ploima are also supported by the data. The relationships within Ploima remain unstable to parameter variation or to the method of phylogeny reconstruction and poorly supported, and the analyses showed that monophyly was questionable for the families Dicranophoridae, Notommatidae, and Brachionidae, and for the genus Proales. Otherwise, monophyly was generally supported for the represented ploimid families and genera. PMID- 16690328 TI - Species sympatry and horizontal transfers of Mariner transposons in marine crustacean genomes. AB - Mariner-like elements (MLEs) have been widely detected in terrestrial species. The first complete MLE isolated from a marine invertebrate was detected in the genome of the hydrothermal crab Bythograea thermydron by Halaimia-Toumi et al. [Halaimia-Toumi, N., Casse, N., Demattei, M.V., Renault, S., Pradier, E., Bigot, Y., Laulier, M., 2004. The GC-rich transposon Bytmar1 from the deep-sea hydrothermal crab, Bythograea thermydron, may encode three transposase isoforms from a single ORF. J. Mol. Evol. 59, 747-760] and called Bytmar1. Here, we report the isolation of three new Bytmar1 relatives from the genomes of one hydrothermal amphipod Ventiella sulfuris (Vensmar1) and two coastal crustacea, Maia brachydactila (Maibmar1) and Cancer pagurus (Canpmar1). Like Bytmar1, these MLEs have an unusually high GC content, a high CpG ratio, and a low TpA ratio. Their consensus sequence encodes a transposase that is preceded by an N-flag, as in Bytmar1, which could be a marine feature. Only one of the 19 clones obtained, Vensmar1.3, encoded for a full-length transposase. The phylogenetic analyses revealed that all these Bytmar1-related elements can be differentiated into two clusters, corresponding to the coastal or hydrothermal origin of their hosts. They also confirmed that the irritans sub-family comprises at least four lineages that seem to depend on the taxonomical position and habitat of their hosts. Finally, we observed that elements coding for two potentially complete transposases exhibiting 99.5% similarity, Bytmar1.11 and Vensmar1.3, were present in the genome of two distantly related hydrothermal crustacea, one Amphipod and one Decapod. The hypothesis of horizontal transfers is discussed in the light of the sequence similarities observed. PMID- 16690329 TI - Depopulated bovine ureteric xenograft for complex haemodialysis vascular access. PMID- 16690330 TI - Can statins reduce perioperative morbidity and mortality in patients undergoing non-cardiac vascular surgery? AB - AIMS: To determine whether statins can reduce perioperative morbidity and mortality in patients undergoing non-cardiac vascular surgery. METHODS: A search using Pubmed was performed to identify reports in English. The search terms were: "statins", "perioperative morbidity", "perioperative mortality" and "vascular surgery". We excluded studies dealing with the effect of statins in cardiac surgery. Retrieved articles were manually searched. RESULTS: Current evidence shows that statins decrease perioperative morbidity and mortality in patients undergoing non-cardiac vascular surgery. Any benefit probably occurs soon (within a month) after initiating treatment. CONCLUSIONS: Appropriately designed trials need to confirm the beneficial effect of perioperative statin therapy in various patient categories. The optimal duration and dose of perioperative statin therapy should be defined. PMID- 16690331 TI - A novel Alu-mediated 61-kb deletion of the von Willebrand factor (VWF) gene whose breakpoints co-locate with putative matrix attachment regions. AB - BACKGROUND AND OBJECTIVES: von Willebrand disease (VWD) type 3 is characterized by extremely low levels of von Willebrand factor (VWF) in plasma. To date, only 11 examples of gross deletions have been reported for the VWF gene and the underlying mutational mechanisms remain unclear. A Chinese patient with type 3 VWD was studied to elucidate the underlying mechanism of mutagenesis. DESIGN AND METHODS: PCR was designed to amplify across the putatively deleted region of genomic DNA from the patient and his parents to locate the deletion breakpoints. In silico analysis was then performed to search for repetitive sequence elements, recombination-associated motifs, and scaffold/matrix attachment regions (S/MARs). RESULTS: A novel homozygous gross deletion of the VWF gene, which removes some 61044 bp DNA between introns 5 and 16, was identified in the patient. The deletion junctions were flanked by highly homologous Alu repeats in inverted orientation. These repeats could thus have potentiated the formation of a stem loop structure thereby bringing the breakpoints into close proximity. A number of recombination-associated motifs were noted in close proximity to both deletion breakpoints. Both the 5' and 3' breakpoints were located in, or near, regions with a high propensity to form S/MARs. INTERPRETATION AND CONCLUSIONS: We report the first example of an Alu-mediated VWF gross gene deletion. Since a number of recombination-associated motifs were also identified in the vicinity of the breakpoints, it may be that multiple sequence elements have acted in concert to give rise to this deletion event. PMID- 16690332 TI - Dynamics of NO rebinding to the heme domain of NO synthase-like proteins from bacterial pathogens. AB - Some Gram-positive bacterial pathogens harbor a gene that encodes a protein (HNS, Heme domain of NO Synthase-like proteins) with striking sequence identity to the oxygenase domain of mammalian NO synthases (NOS). However, they lack the N terminal and the Zn-cysteine motif participating to the stability of an active dimer in the mammalian isoforms. The unique properties of HNS make it an excellent model system for probing how the heme environment tunes NO dynamics and for comparing it to the endothelial NO synthase heme domain (eNOS(HD)) using ultrafast transient spectroscopy. NO rebinding in HNS from Staphylococcus aureus (SA-HNS) is faster than that measured for either Bacillus anthracis (BA-HNS) or for eNOS(HD) in both oxidized and reduced forms in the presence of arginine. To test whether these distinct rates arise from different energy barriers for NO recombination, we measured rebinding kinetics at several temperatures. Our data are consistent with different barriers for NO recombination in SA-HNS and BA-HNS and the presence of a second NO-binding site. The hypothesis that an additional NO-binding cavity is present in BA-HNS is also consistent with the effect of the NO concentration on its rebinding. The lack of the effect of NO concentration on the geminate rebinding in SA-HNS could be due to an isolated second site. We confirm the existence of a second NO site in the oxygenase domain of the reduced eNOS as previously hypothesized [A. Slama-Schwok, M. Negrerie, V. Berka, J.C. Lambry, A.L. Tsai, M.H. Vos, J.L. Martin, Nitric oxide (NO) traffic in endothelial NO synthase. Evidence for a new NO binding site dependent on tetrahydrobiopterin? J. Biol. Chem. 277 (2002) 7581-7586]. This site requires the presence of arginine and BH(4); and we propose that NO dynamic and escape from eNOS is regulated by the active site H-bonding network connecting between the heme, the substrate, and cofactor. PMID- 16690333 TI - Brain abscess in a dog immunosuppressed using cyclosporin. AB - This report describes a dog that developed brain abscesses following prolonged immunosuppression with cyclosporin. Bacteria within the abscess were most likely Nocardia, an organism well recognised in immunosuppressed humans, and probably reached the brain through haematogenous spread from a more long-standing abscess in the mediastinum. Bacterial brain abscesses developing in this manner are very rare in dogs and this case highlights the wider range of possible diagnoses that need to be considered in immunosuppressed patients and the care with which potent drugs such as cyclosporin should be used. PMID- 16690334 TI - Molecular profiling and comparison of field transmissible spongiform encephalopathy cases diagnosed in Catalunya. AB - Molecular profiling of the proteinase K resistant prion protein (PrP(res)) is a technique that has been applied to the characterisation of transmissible spongiform encephalopathy (TSE) strains. An interesting example of the application of this technique is the ability to differentiate, at the experimental level, between bovine spongiform encephalopathy (BSE) and scrapie infection in sheep, and to distinguish between classical and atypical BSE and scrapie cases. Twenty-six BSE cases and two scrapie cases from an active TSE surveillance program and diagnosed at the PRIOCAT, TSE Reference Laboratory (Centre de Recerca en Sanitat Animal, Universitat Autonoma de Barcelona, Catalunya, Spain) were examined by Western blotting. Molecular profiling was achieved by comparing the glycosylation profile, deglycosylated PrP molecular weight and 6H4/P4 monoclonal antibody binding ratio. The results obtained during the characterisation of these field cases indicated an absence of atypical BSE cases in Catalunya. PMID- 16690335 TI - Comparative analysis of haematological, haemostatic, and inflammatory parameters in canine venous and arterial blood samples. AB - The objective of the present study was to validate the use of blood collected from an indwelling arterial catheter for analysis of haematological, coagulation and inflammatory parameters in canines compared to venous blood collected directly from the jugular vein. Blood samples were collected from 11 dogs. Agreement between sampling methods was found for neutrophil and monocyte counts, prothrombin time, activated partial thromboplastin time, antithrombin, protein C, factor VIII and C-reactive protein, whereas a statistically significant difference was found for white blood cells, lymphocyte, erythrocyte and platelet counts, haemoglobin, haematocrit, fibrinogen and thrombin time (TT). In conclusion, it is necessary to be aware that results from a complete blood count obtained from canine venous and arterial blood samples may not be comparable. Values for haemostatic parameters from arterial and venous blood samples, with the exception of fibrinogen and TT, were however statistically identical. PMID- 16690336 TI - Changes in dorsal root ganglion CGRP expression in a chronic inflammatory model of the rat knee joint: differential modulation by rofecoxib and paracetamol. AB - Neuropeptide-expressing small diameter sensory neurones are thought to be vital in generating inflammatory hyperalgesic responses. Within the dorsal root ganglion (DRG), both the levels of the neuropeptide calcitonin gene-related peptide (CGRP) and the numbers of CGRP-immunoreactive (CGRP-IR) DRG neurones have been shown to increase in a number of acute adjuvant-induced inflammatory pain models. The aim of this study was to look specifically at changes in numbers of CGRP-IR DRG neurones in a chronic model of inflammatory joint pain following complete Freund's adjuvant (CFA) injection into the rat knee. In this model, there were significant increases in the number of ipsilateral CGRP-IR small DRG neurones at days 1, 16 and 35 following intra-articular CFA, compared to saline injected sham animals. This correlated with the behavioural readouts of hypersensitivity and knee joint inflammation at the same time points. There was also a significant increase in the number of ipsilateral CGRP-IR medium DRG neurones and contralateral CGRP-IR small DRG neurones at day 1. Following dosing of CFA-injected rats with rofecoxib (Vioxx) or paracetamol, there was a significant decrease in the number of ipsilateral CGRP-IR small and medium DRG neurones in rofecoxib- but not paracetamol-treated rats. These data also correlated with behavioural readouts where hypersensitivity and knee joint inflammation were significantly reduced by rofecoxib but not paracetamol treatment. In conclusion, these data show that changes in ipsilateral CGRP expression within small DRG neurones are consistent with behavioural readouts in both time course, rofecoxib and paracetamol studies in this model of chronic inflammatory pain. PMID- 16690337 TI - Effect of nitrogen dioxide on the EPR property of lithium octa-n-butoxy 2,3 naphthalocyanine (LiNc-BuO) microcrystals. AB - Lithium octa-n-butoxy-naphthalocyanine (LiNc-BuO) is a stable free radical that can be detected by electron paramagnetic resonance (EPR) spectroscopy. Previously we have reported that microcrystals of LiNc-BuO exhibit a single sharp EPR peak, whose width varies linearly with the partial pressure of paramagnetic molecules such as oxygen and nitric oxide. In this report, we present the effect of nitrogen dioxide (NO2), which is also a paramagnetic molecule, on the EPR properties of LiNc-BuO. The gas-sensing property of LiNc-BuO is attributed to the open molecular framework of the crystal structure which is arranged with wide channels capable of accommodating large molecules such as NO2. The EPR linewidth of LiNc-BuO was highly sensitive to the partial pressure of NO2 in the gas mixture. The line-broadening was quick and reversible in the short-term for low concentration of NO2. However, the EPR signal intensity decreased with time of exposure, apparently due to a reaction of NO2 with LiNc-BuO crystals to give diamagnetic products. The results suggested that LiNc-BuO may be a useful probe for the determination of trace amounts of NO2 using EPR spectroscopy. PMID- 16690338 TI - Beginning to end: cardiovascular implications of growth hormone (GH) deficiency and GH therapy. AB - Both growth hormone (GH) and insulin-like growth factor I (IGF-I) are involved in heart development and in maintenance of cardiac structure and performance. Cardiovascular disease has been reported to reduce life expectancy in both GH deficiency (GHD) and GH excess. Patients with GHD suffer from a cluster of abnormalities associated with increased cardiovascular risk, including abnormal body composition, unfavorable lipid profile, increased fibrinogen and C-reactive protein levels, insulin resistance, early atherosclerosis and endothelial dysfunction, and impaired left ventricular (LV) performance (i.e., reduced diastolic filling and impaired response to peak exercise). Long-term GH replacement therapy reverses most of these abnormalities. More consistently, GH replacement reduces body fat and visceral adipose tissue, reduces low-density lipoprotein cholesterol and triglyceride levels, and improves endothelial function. GH replacement also reduces intima media thickness at major arteries and improves LV performance, but these results have been observed only in small series of patients treated on a short-term basis. This review discusses the roles of GHD and GH replacement therapy in the development of cardiovascular disease. PMID- 16690339 TI - Endocrinology--the way we were: a personal history of somatomedin. AB - Fifty years ago there was no practical method for measuring growth hormone (GH) activity in human plasma. In an attempt to develop such an assay, it was determined that much of the action of GH in vivo on rat cartilage was mediated by a serum factor that was first known as sulfation factor, and later as somatomedin and insulin-like growth factor. This narrative describes my experience and that of others in this important area of endocrine research. PMID- 16690340 TI - The impact of genomics on discovering drugs against infectious diseases. AB - Genomics is accelerating the progress in data generation and interpretation in the global analyses of components of cells, including the spectrum of lipids, RNA, metabolites, proteins, mutational phenotypes or DNA methylation sites. Integration of the knowledge generated by these diverse strategies is predicted to have a tremendous impact on approaches to rational drug discovery against infectious diseases. PMID- 16690341 TI - Cardiovascular risk and rheumatoid arthritis: clinical practice guidelines based on published evidence and expert opinion. AB - OBJECTIVE: To develop clinical practice guidelines for the evaluation and management of cardiovascular risk in patients with rheumatoid arthritis (RA), using the evidence-based approach and expert opinion. METHODS: Recommendations were developed using the evidence-based approach and expert opinion: A scientific committee used a Delphi procedure to select five questions, which formed the basis for developing the recommendations; Evidence providing answers to the five questions was sought in the literature; Based on this evidence, recommendations were developed by a panel of experts. RESULTS: The recommendations were as follows: 1) In patients with RA, attention should be given to the risk of cardiovascular disease, which is responsible for an excess burden of morbidity and mortality; 2) It must be recognized that RA may be an independent cardiovascular risk factor. Persistent inflammation is an additional risk factor; 3) The cardiovascular risk should be evaluated, and modifiable risk factors should be corrected; 4) In patients with RA requiring glucocorticoid therapy, the need for cardiovascular risk minimization is among the reasons that mandate the use of the minimal effective dose; 5) It should be recognized that methotrexate may protect against cardiovascular mortality in patients with RA; 6) It should be recognized that TNFalpha antagonists remain contraindicated in patients with RA and severe heart failure. TNFalpha antagonists do not seem to worsen moderate heart failure and may protect against cardiovascular mortality; 7) AFSSAPS recommendations about LDL-cholesterol objectives should be followed, with active RA being counted as a cardiovascular risk factor; 8) In patients with RA, statin therapy should be considered only when cholesterol levels are elevated despite appropriate dietary treatment; 9) RA per se does not indicate aspirin for primary prevention. When aspirin is used for secondary prevention, it should be recognized that concomitant treatment with nonsteroidal antiinflammatory drugs (NSAIDs) may decrease the antiplatelet effects and increase the gastrointestinal side effects of aspirin therapy. PMID- 16690342 TI - [Vaginoscopic hysteroscopy]. AB - Vaginoscopic hysteroscopy permits to avoid several painful gestures of classical office hysteroscopy. The aim of this article is to describe the advantages such vaginoscopic approach. Six studies were thus selected, dealing with vaginoscopic hysteroscopy. The diameter of the hysteroscope, always a rigid one, was between 3.5 and 5 mm, and CO(2) or saline infusions were indifferently used. Failure rate is inferior to 5%. One study, comparing pain induced by vaginoscopic versus classical hysteroscopy, concluded that vaginoscopic approach was less painful. PMID- 16690343 TI - Selective hydrogenation of furfural to furfuryl alcohol over catalysts prepared via sonochemistry. AB - Copper-chromite oxide and TiO2-supported copper-chromite oxide catalysts are prepared by various methods. They are characterized with ICP, BET, XRD, XPS, SEM, and TEM, etc. Their catalytic performance for liquid phase hydrogenation of furfural to furfuryl alcohol is also valuated. The catalysts prepared by ultrasound exhibit good performance. Catalytic activity of TiO2-supported catalysts is higher than that of catalyst without TiO2, notwithstanding they are all prepared by ultrasound. It is worth stressing that after reduced the TiO2 supported catalysts, which are X-ray amorphous, display good performance at 140 degrees C, while the catalysts without TiO2 show no activity under the same condition. Obtained results indicate that the catalytic performance of catalysts depends upon the amount of reducible copper ions and the activity decay is related to the loss of metal elements on the surface of catalyst. PMID- 16690344 TI - Cyclic hydrostatic pressure and particles increase synthesis of 1,25 dihydroxyvitamin D3 by human macrophages in vitro. AB - 1,25-Dihydroxyvitamin D(3) has a pivotal role in bone resorption and osteoclast activity. As activated macrophages are known to synthesise 1,25-dihydroxyvitamin D(3), this study examined whether pressure modulated its synthesis. Pressure and particles have been shown to increase synthesis of pro-resorptive cytokines and other factors by cultured macrophages. Human peripheral blood macrophages were isolated, cultured and exposed to pressure (similar to that found in the human joint) and/or particles. Synthesis of 1,25-dihydroxyvitamin D(3) by macrophages was assayed using high pressure liquid chromatography and in situ hybridization. Synthesis of 1,25-dihydroxyvitamin D(3) but not 24,25-dihydroxyvitamin D(3) was increased in macrophages under pressure. In situ hybridization demonstrated an increase in 1alpha-hydroxylase expression in response to pressure or particles and simultaneous exposure to both stimuli generated higher expression of 1alpha hydroxylase. In conclusion, this is the first study to demonstrate that mechanical loading, in the form of pressure, stimulates 1,25-dihydroxyvitamin D(3) synthesis in human macrophages. These findings have implications for the in vivo situation, as they suggest that 1,25-dihydroxyvitamin D(3) could be one factor stimulating osteoclastic bone resorption in pathologies, such as arthritis or implant loosening, where intra-articular or intra-osseous pressure is raised or where wear particles interact with macrophages. PMID- 16690345 TI - History, protohistory and prehistory of the Arabidopsis thaliana chromosome complement. AB - We propose an evolutionary scenario that could have shaped the modern Arabidopsis thaliana genome, which began with the reduction in chromosome number from n=8 to n=5 in the past 4 million to 5 million years as a result of chromosome fusion. The scenario also includes three ancient polyploidizations: the most recent occurred in an early Brassicaceae with n=4 chromosomes 24 million to 40 million years ago. The two other polyploidizations occurred after the emergence of the Eudicots and the Angiosperms, respectively. Angiosperm evolution includes recurrent cycles of genome duplication and gene and chromosome reorganizations. PMID- 16690346 TI - Recent developments and future prospects in insect pest control in transgenic crops. AB - The adoption of insect-resistant transgenic crops has been increasing annually at double-digit rates since the commercial release of first-generation maize and cotton expressing a single modified Bacillus thuringiensis toxin (Bt) nine years ago. Studies have shown that these Bt crops can be successfully deployed in agriculture, which has led to a decrease in pesticide usage, and that they are environmentally benign. However, the sustainability and durability of pest resistance continues to be discussed. In this review, we focus on the science that underpins second- and third-generation insect-resistant transgenic plants and examine the appropriateness and relevance of models that are currently being used to determine deployment strategies to maximize sustainability and durability. We also review strategies that are being developed for novel approaches to transgenic insect pest control. PMID- 16690347 TI - Clinical performance of daily disposable soft contact lenses using sustained release technology. AB - PURPOSE: Polyvinyl alcohol (PVA) is a successful tear film stabiliser and is widely used in comfort drops and some soft contact lens materials. A PVA containing lens, nelfilcon A has been modified to include additional (non functional) PVA in order to provide improved comfort. This study aims to examine the clinical performance of this nelfilcon A lens with AquaRelease (AquaRelease). METHODS: Two contralateral, investigator masked, open label, subjective and objective evaluations were conducted. The first examined the effect of adding increased molecular weight PVA to nelfilcon A (n=5), and the second compared this AquaRelease lens to ocufilcon B (n=34). The principal measures were non-invasive break-up time (NIBUT) and subjective comfort, which were assessed at the beginning and end of a week of daily wear, and three times throughout 1 day at 8, 12 and 16 h. RESULTS: All subjects successfully completed the daily wearing schedule of 16h. On initial insertion, subjective comfort and NIBUT improved for AquaRelease than original nelfilcon A lenses (p<0.05). Initial comfort was better for AquaRelease compared to ocufilcon B lenses (p=0.01); however, NIBUT was not statistically different (11.7+/-15.6s versus 8.4+/-6.8s; p=0.26). Subjective comfort decreased with time (p<0.001), but there was no significant difference between AquaRelease and ocufilcon B lenses (p=0.16). NIBUT was not significantly affected by time (p=0.56) or between lenses (p=0.33). At the end of a weeks' wear, subjective initial, end-of-day, overall comfort and vision were rated significantly better with AquaRelease than ocufilcon B (p<0.01). CONCLUSIONS: Release of additional non-functionalised PVA from the nelfilcon A lenses appears to enhance comfortable contact lens wear. PMID- 16690349 TI - Correlation between induction of DNA fragmentation in lung cells from rats and humans and carcinogenic activity. AB - Six chemicals, known to induce lung tumors in rats, were examined for their ability to induce DNA fragmentation in primary cultures of rat and human lung cells, and in the lung of intact rats. Significant dose-dependent increases in the frequency of DNA single-strand breaks and alkali-labile sites, as measured by the single-cell gel electrophoresis (Comet) assay, were obtained in primary lung cells from male rats with the following, minimally toxic, concentrations of the six test compounds: N-nitrosodimethylamine (NDMA; 2.5-10 mM), hydrazine (HZ; 0.5 4 mM), cadmium sulfate (CD; 31.2 and 62.5 MUM), 4,4'-methylene bis (2 chloroaniline) (MOCA; 31.2-125 MUM), isobutyl nitrite (IBN; 7.8-31.2 MUM) and tetranitromethane (TNM; 1.9-15.6 MUM). Similar degrees of DNA fragmentation were obtained in primary human lung cells; however, due to inter-donor differences, the minimum effective concentrations were in some donors lower and in others higher than in rats, and IBN induced DNA damage only in one of three donors. The DNA-damaging potency of HZ was higher in rats than in humans, and the opposite was true for MOCA. In agreement with these findings, statistically significant increases in the average frequency of DNA breaks were obtained in the lung of rats given a single oral dose (1/2 LD50) of the six test compounds. These findings give evidence that genotoxic lung carcinogens may be identified by use of the DNA fragmentation/Comet assay on rat lung cells as targets cells, and show that the six compounds tested produce in primary cultures of lung cells from human donors DNA-damaging effects substantially similar to those observed in rats. PMID- 16690350 TI - Evaluation of high-risk human papillomavirus types PCR detection in paired urine and cervical samples of women with abnormal cytology. AB - BACKGROUND: During the last decade, increasing efforts have focused on HPV detection in self-obtained samples, to increase the overall proportion of patients participating in cervical cancer screening procedures. OBJECTIVES: A clinical evaluation study of an optimized protocol for PCR detection of high-risk human papillomavirus (HPV) types in urine compared with cervical samples in consecutive women referred to the colposcopy clinic with abnormal cervical cytology. STUDY DESIGN: Paired urine and cervical specimens were collected from 100 consecutive women referred to the colposcopy clinic with abnormal cervical cytology and normal urine parameters. In-house and a commercial PCR method for the detection of HPV types 16 and 18, and a commercial multiplex PCR for HPV types 6, 11, 16, 18, and 33 were performed. All HPV cervix-positive/urine negative paired urine samples were spiked with serial dilutions of cell lines infected with HPV 16 or 18 to test the sensitivity of HPV detection in these urine samples. RESULTS: In all but two cases HPV type 16 was detected. In cancer cases, the urine/cervix HPV detection sensitivity was 88.8%; in cases with high grade lesions it was 76.5%; and in cases with low-grade lesions it was 45.5%. In all concordant cases the same HPV type was detected in both samples. The urine/cervix HPV detection sensitivity was higher when urine samples contained two or more epithelial cells per field in urine microscopy. HPV detection in 9 cervix-positive but urine-negative urine samples spiked with serial dilutions of HPV-positive cell lines showed that in these cases urine PCR inhibitors did not affect PCR amplification. CONCLUSIONS: A higher urine/cervix HPV detection sensitivity in cancer and high-grade lesions suggests that urine testing could be used to detect HPV mainly when these lesions are present. PMID- 16690351 TI - Neuronal function in chronic spinal cord injury: divergence between locomotor and flexion- and H-reflex activity. AB - OBJECTIVE: Knowledge about the long-term course of spinal neuronal function after spinal cord injury (SCI) is important if regeneration therapies become available in the future. The objective of this study was to examine the behavior of locomotor EMG activity and of spinal reflexes in patients with chronic motor complete SCI. METHODS: EMG activity from rectus femoris (RF), biceps femoris (BF), medial gastrocnemius (GM) and tibialis anterior (TA) of both sides was investigated during locomotor movements assisted by a robotic device in 10 chronic (>1 year after accident) complete SCI and 5 healthy subjects. H-reflexes (recorded from GM) were induced during the onset and flexion reflexes (recorded from BF and TA) at the end of the stance phase. RESULTS: Only in the chronic SCI subjects an exhaustion of EMG activity--i.e. a decrease in amplitude--occurred within a few minutes in all leg muscles. The EMG exhaustion was not associated with a change in the H- or flexion reflex amplitude during a walking session. CONCLUSIONS: Exhaustion of neuronal function in chronic complete SCI might be restricted to unused motor tasks, i.e. locomotion. The fact that H- and flexion reflexes show a normal behavior might be due to the fact that they still become activated. SIGNIFICANCE: Training/pharmalogical approaches may be required to maintain neuronal function of unused tasks as a basis for future successful regeneration therapies. PMID- 16690352 TI - Fatigue and anaemia in patients with chronic heart failure. AB - BACKGROUND: Fatigue is common in patients with chronic heart failure (CHF) and has great impact on functional ability and daily activity. Although anaemia is associated with fatigue, the relationship between fatigue and anaemia is unclear in CHF. The aim of this study was to describe the fatigue experience and its relationship to haemoglobin (Hb) concentration and to evaluate its effect on health-related quality of life in an unselected hospitalised CHF population. METHODS: Ninety three consecutive patients hospitalised with a diagnosis of CHF, enrolled in the EuroHeart Failure Survey, completed the Multidimensional Fatigue Inventory Scale (MFI-20). New York Heart Association (NYHA) functional class, quality of life and haemoglobin were also assessed. RESULTS: Anaemia (Hb < or = 125 g/L) was found in 31 (33%) patients. The perception of fatigue differed significantly between patients with CHF and healthy individuals. Anaemic patients reported significantly more fatigue compared to non-anaemic patients. Decreased haemoglobin and higher NYHA class explained 30% of the variance in General Fatigue. Perceived fatigue was also inversely related to global health and quality of life. CONCLUSION: Our findings suggest that the subjective experience of fatigue in patients with CHF is associated with low haemoglobin concentration and reduced functional status after controlling for age and sex. PMID- 16690353 TI - Effects of high-intensity interval training on the VO2 response during severe exercise. AB - This study examined the effect of high-intensity interval training on the VO2 response during severe, constant-load exercise. Prior to, and following training, 10 females (V O2 peak 37.4+/-6.0 mL kg-1 min-1) performed a graded exercise test to determine VO2 peak and lactate threshold (LT) and a 6 min cycle test (CT) at the pre-training VO2 peak intensity. Training involved high-intensity intervals (2 min work, 1 min rest) performed 3x week for 8 weeks. Breath-by-breath data from 0 to 6 min during the CT were smoothed using 5s averages and fit to a bi exponential model starting from 20s. Training resulted in significant improvements in VO2 max (2.34+/-0.37-2.78+/-0.30 L min-1), power at VO2 max (170+/-26-204+/-25 W) and power at LT (113+/-17-136+/-20 W) (p<0.05). Following training, the VO2 response showed a significant increase in the amplitude of the primary phase (A1) (1396+/-103-1695+/-100 mL min-1; p<0.05) and end-exercise VO2 (VO2 EE), with no difference (p>0.05) in the time constants of either phase or the amplitude of the slow component (318+/-67-380+/-48 mL; p=0.15). In conjunction, accumulated oxygen deficit (AOD) (43.7+/-9.8-17.2+/-2.8 mL O2 eq kg 1) and anaerobic contribution to the CT (19.4+/-4.4-7.2+/-1.2%) were significantly reduced. In contrast to previous moderate-intensity research, a high-intensity interval training program increased A1 and VO2 EE for the same absolute exercise intensity, decreasing the AOD during a severe-intensity CT. PMID- 16690354 TI - A novel multi-planed mechanical aortic valve for increasing the effective orifice area. AB - BACKGROUND: Today, there is still confusion about mismatch of the orifice area of the mechanical valve and body surface area of the patient in aortic valve surgery. Creating a larger effective orifice area is the aim with this new valve design. METHOD: This valve is multi-planed, one housing is seated at the aortic annulus for the coronary orifices to receive blood in diastole, the other housing or housings are seated to the ascending aorta obliquely to increase the orifice area of the valve. The ascending aorta can be enlarged if necessary. RESULT: Valves with orifice areas larger than 6 cm(2) can be achieved with multi-planed aortic valves. DISCUSSION: The use of this valve depends on the fact that the aorta is a living tissue and can grow over time to normal values. PMID- 16690355 TI - Modulation of multidrug resistance efflux pump activity to overcome chemoresistance in cancer. AB - Early publications using cultured cancer cells immediately recognized the phenomenon of resistance to anticancer agents. However, it was not until 1973 that it was first demonstrated that a major factor in the resistance of cancer cells was that of reduced drug accumulation. This year marks the 30th anniversary of the discovery by Juliano and Ling that P-glycoprotein mediates this active efflux of chemotherapeutic drugs from cancer cells. Since this seminal finding, the investigation of P-glycoprotein (MDR1, ATP binding cassette [ABC]B1) has proceeded with great vigour. However, it soon became apparent that P-glycoprotein was not expressed in all drug-resistant cells that displayed an accumulation deficiency, which led to the discovery of other ABC transporters involved in drug efflux. In 1992, the multidrug resistance-associated protein (MRP1, ABCC1) was identified in small cell lung cancer followed by breast cancer resistance protein (mitoxantrone resistance protein, ABCG2) in 1999. After three decades of research, can we confidently define the contribution of multidrug resistance transporters to chemoresistance and do we have clinically useful drugs to sensitise cancers? PMID- 16690356 TI - New insights into the pathophysiology of cobalamin deficiency. AB - Cobalamin-deficient (Cbl-D) central neuropathy in the rat is associated with a locally increased expression of neurotoxic tumour necrosis factor-alpha (TNF alpha) and a locally decreased expression of neurotrophic epidermal growth factor (EGF). These recent findings suggest that cobalamin oppositely regulates the expression of TNF-alpha and EGF, and raise the possibility that these effects might be independent of its coenzyme function. Furthermore, adult Cbl-D patients have high levels of TNF-alpha and low levels of EGF in the serum and cerebrospinal fluid. Serum levels of TNF-alpha and EGF of cobalamin-treated patients normalize concomitantly with haematological disease remission. These observations suggest that cobalamin deficiency induces an imbalance in TNF-alpha and EGF levels in biological fluids that might have a role in the pathogenesis of the damage caused by pernicious anaemia. PMID- 16690357 TI - Implementation of national schistosomiasis control programmes in West Africa. AB - Burkina Faso, Mali and Niger are countries endemic for schistosomiasis, with a high predominance of Schistosoma haematobium. With the support of the Bill and Melinda Gates Foundation through the Schistosomiasis Control Initiative, national control programmes were launched in these countries in 2004. Here, we describe the progress of implementation for each programme and the challenges for maintaining sustainability for schistosomiasis control in these countries. PMID- 16690358 TI - Evaluation of the shortened Cholesterol-Lowering Diet Self-Efficacy Scale. AB - Specificity in the self-efficacy construct requires that instruments assess domain-specific behaviors. The objectives of the study were to (1) reexamine and shorten the original 57-item Cholesterol-Lowering Diet Self-Efficacy Scale (CLDSES), (2) estimate reliability and validity of the short form CLDSES (CLDSES SF), (3) examine the dimensionality of the CLDSES-SF, (4) examine discriminant validity of the scale by its ability to differentiate between adherence and nonadherence to saturated fat intake guidelines, and (5) examine the CLDSES-SF's sensitivity and specificity. A sample of 238 patients being treated for hypercholesterolemia completed three instruments: the CLDSES, the Connor Diet Habit Survey (DHS) and a Three-Day Food Record. Internal consistency for the CLDSES-SF was .95. The inter-correlation coefficients among the seven sub-scales ranged from .28 to .62, and for the total to sub-scales .58 - .87, significant at p < .001. Evidence for criterion validity was shown in the correlations between the CLDSES-SF and the Connor DHS scores and between the CLDSES-SF and the Three Day Food Record. Factor analysis showed a one-factor model that fits the data with a sufficient amount of total variance explained among the seven subscale items in each subscale, providing evidence of the uni-dimensionality of each subscale. Sensitivity (76%) and specificity (63%) for the CLDSES-SF were good with 88% positive predictive value for adherence. The shortened CLDSES retained its good psychometric properties and remains comprehensive in its assessment of self-efficacy for adherence to a cholesterol-lowering diet. PMID- 16690359 TI - A phase I trial of intravenous CG7870, a replication-selective, prostate-specific antigen-targeted oncolytic adenovirus, for the treatment of hormone-refractory, metastatic prostate cancer. AB - CG7870 is a replication-selective oncolytic adenovirus genetically engineered to replicate preferentially in prostate tissue. In a previous phase I/II clinical trial of intraprostatic delivery of CG7870 for locally recurrent prostate cancer this virus was well tolerated. In this phase I study CG7870 was administered as a single intravenous infusion in a group-sequential dose escalation design (1 x 10(10) to 6 x 10(12) viral particles (vp)) to 23 patients with hormone-refractory metastatic prostate cancer. Flulike symptoms (fever, fatigue, rigors, nausea, and/or vomiting) were the most common adverse events. Three therapy-related grade 3 adverse events were reported, one of which (fatigue) was serious. At doses greater than 10(12) vp all five patients experienced asymptomatic grade 1 to 2 transaminitis and/or isolated d-dimer elevations starting on day 2 through 8; dose escalation was therefore halted at 6 x 10(12) vp. All tested patients had CG7870 genomes present in the peripheral blood for at least 90 minutes after infusion; patients in the highest dose group had persistence of genomes through 29 days. A "secondary" or "delayed" peak in plasma CG7870 genome copies (defined as a >10-fold increase in CG7870 genomes from nadir concentration) suggestive of active viral replication and shedding into the bloodstream was detected in 16/23 (70%) patients. CG7870 was detected in the saliva of 3 patients, whereas all urine samples tested negative. All patients developed antibodies to CG7870. Dose related increases in interleukins 6 and 10 (IL-6, IL-10) blood levels were detected. The peak IL-6 concentration after CG7870 treatment was associated with a transient, asymptomatic decrease in blood pressure. No partial or complete prostate-specific antigen (PSA) responses were observed; however, 5 patients had a decrease in serum PSA of 25% to 49% following a single treatment, including 3 of 8 patients at the highest dose levels. PMID- 16690360 TI - Epilepsy and workplace discrimination: population characteristics and trends. AB - This study investigates the nature of allegations of workplace discrimination made by job seekers and workers with epilepsy. Demographic characteristics of individuals making allegations were consistent with those of individuals with epilepsy nationwide. Approximately three-fourths of allegations were related to post-hire issues, such as discrimination in promotion and termination, disciplinary actions, and harassment. The lowest rate of merit allegation was related to discrimination in hiring. Comparison of merit allegations before and after the U.S. Supreme Court's Sutton decision revealed that claimants have had higher rates of success following the decision, suggesting that the Sutton decision has had no negative effect on claimants with epilepsy. PMID- 16690361 TI - Field-based evidence for linkage of mutations associated with chloroquine (pfcrt/pfmdr1) and sulfadoxine-pyrimethamine (pfdhfr/pfdhps) resistance and for the fitness cost of multiple mutations in P. falciparum. AB - Mutations in the Plasmodium falciparum pfcrt gene on chromosome 7 and possibly mutations in pfmdr1 on chromosome 5 have a role in conferring resistance against chloroquine (CQ), as do mutations of pfdhfr on chromosome 4 and pfdhps on chromosome 8 in terms of resistance against sulfadoxine/pyrimethamine (SP). The additive role of multiple mutations in the development of resistance to each drug suggests a non-random occurrence. In this study, parasite isolates were obtained from 50 patients with uncomplicated P. falciparum malaria from rural Eastern Sudan, an endemic setting with minimal overlap of infection. The parasite isolates were genotyped for detection of 12 alleles in CQ and SP resistance genes. Our main findings were: (1) the frequency of mutant alleles, pfcrt K76T, pfmdr1 N86Y, pfdhfr N51I, pfdhfr S108N, pfdhps K540E and pfdhps A581G were; 0.90, 0.86, 0.84, 0.84, 0.80 and 0.20, respectively. (2) No mutations were detected for the pfdhfr loci A16V, C59R and I164L, and for pfdhps loci S436A, A437G and A613S. (3) There was a statistically significant association between the mutations in: (i) the CQ resistance (CQR) genes, pfcrt T76 and pfmdr1 Y86 (P< or =0.001), (ii) the SP resistance (SPR) genes, pfdhfr I51, pfdhfr N108 and pfdhps E540 (P< or =0.001-0.04) and (iii) the CQ "i" and SP "ii" resistance genes (P=0.001) 4. The fitness cost of multiple mutations was revealed by a significantly reduced parasite density of isolates bearing the mutant alleles (P=0.048). However, the significantly higher gametocyte carriage rate among isolates with resistance mutations (P=0.001) is possibly an evolutionary mechanism for survival of mutant parasites. PMID- 16690362 TI - Proceedings of the 4th International Symposium on Separations in the Biosciences. September 18-21, 2005. Utrecht, The Netherlands. PMID- 16690363 TI - Intestinal immune defences and the inflammatory response in necrotising enterocolitis. AB - Necrotising enterocolitis is a devastating neonatal gastrointestinal emergency predominantly affecting low birth weight, premature infants and is accompanied by significant mortality and morbid sequelae. The pathophysiology remains obscure and the management of infants with necrotising enterocolitis has not changed since the recognition of this disease. Necrotising enterocolitis is most likely the clinical culmination of multiple different risk factors interacting with each other to produce bowel injury through a final, common inflammatory pathway. Here, we review intestinal immunity and the specific inflammatory mediators involved in this disease process. PMID- 16690364 TI - SIRS and group-B streptococcal sepsis in newborns: pathogenesis and perspectives in adjunctive therapy. AB - Clinical signs of systemic inflammation and suspected systemic infection are common in neonatal medicine. Yet, causative infectious organisms can only infrequently be isolated. In previously healthy infants at low risk of sepsis, group B streptococcus (GBS) is the most common isolate. In vitro and in vivo data suggest that immune cells from newborn infants have impaired antimicrobial properties against GBS. In contrast large amounts of inflammatory mediators are formed upon GBS challenge and Toll-like receptors (TLR) are critical host molecules in this context. Thus, the immune balance tilts towards inflammation, SIRS and sepsis. Adjunctive therapy of neonatal sepsis needs to adjust the inflammatory response without further impairing bacterial clearance. This article summarises the pathophysiological events leading to sepsis and suggests molecular targets for adjunctive therapy. PMID- 16690365 TI - Type 2 diabetes is not independently associated with spinal trabecular volumetric bone mineral density measured by QCT in the Diabetes Heart Study. AB - The purpose of this study was to investigate the association between type 2 diabetes mellitus (DM2) and trabecular volumetric bone mineral density (vBMD) of the thoracic and lumbar spine measured by quantitative computed tomography (QCT) in 483 female (410 with DM2) and 398 male (365 with DM2) adults (age 36-86 years, BMI 16-58, 88% with DM2) in the Diabetes Heart Study. After accounting for familial correlation using generalized estimating equations (GEE), lumbar spine vBMD was positively associated with BMI (r = 0.24, P < 0.0001) and inversely associated with age (r = -0.51, P < 0.0001). In women, age-adjusted thoracic spinal vBMD (mg/ml, mean +/- SE) was higher in diabetics (147.6 +/- 2.3) compared to unaffected individuals (138.6 +/- 3.4) (P = 0.02), with age-adjusted lumbar spinal vBMD showing a similar but non-significant trend (132.9 +/- 2.1 in diabetics vs. 127.2 +/- 3.6 in unaffected individuals, P = 0.15). In contrast, in men, age-adjusted lumbar and thoracic vBMD were not different between diabetics and unaffected controls (lumbar vBMD = 125.0 +/- 1.8 in diabetics and 125.8 +/- 5.6 in unaffected individuals, P = 0.89; thoracic vBMD = 137.4 +/- 2.1 in diabetics vs. 134.2 +/- 5.5 in controls, P = 0.56). After multivariate analysis adjusting for age, sex, race, BMI, physical activity, dietary intake, smoking, and alcohol use, interaction between diabetes status and trabecular vBMD of the spine was no longer observed. In women only, age-adjusted areal BMD (determined by dual X-ray absorptiometry (DXA)) of the spine and hip were significantly higher in diabetics than non-diabetic (all P < 0.05), although the differences disappeared after additional adjustment for BMI. These data suggest that areal BMD measured by DXA and trabecular volumetric BMD measured by QCT are not associated with type 2 diabetes independently from BMI. PMID- 16690366 TI - Immune system and bone metabolism: Does thymectomy influence postmenopausal bone loss in humans? AB - Recent studies of animal models have suggested that an increase in the number of T cells due to both peripheral expansion and increased thymic T cell output plays a key role in the regulation of bone loss after ovariectomy. Osteoclastogenic cytokines which are either produced by T cells or activate T cells have also been implicated in ovx induced bone loss. Among them are TNF alpha and IL-7. The present study investigates the role of thymectomy (THX) and IL-7 in bone metabolism in humans. We studied T cells subsets, cytokine production and bone metabolism in 13 women thymectomized for Myasthenia gravis as compared to healthy controls. Our data demonstrate that the number of CD4+ and TNF-producing T cells is lower in THX women as compared to euthymic controls. However in THX women the residual T cells produce higher levels of IL-7 and RANKL. Furthermore, flow cytometry shows that IL-7 is produced by T and B cells. Serum levels of TNF alpha were unaffected by THX and both serum TNF alpha and the RANKL/OPG correlated inversely with BMD. There were no differences in bone turnover and bone mineral density between THX women and the controls. These data suggest that THX decreases the number of TNF-producing CD4+ T cells but does not alters serum TNF levels. The RANKL/OPG ratio and indices of bone metabolisms are also not affected by THX, although THX increases the levels of IL-7 and RANKL. Further studies are needed to clarify the role of thymus in bone metabolism and osteoclastogenesis in postmenopausal women. PMID- 16690367 TI - An investigation of epidemiologic factors associated with large nodular goiter. AB - BACKGROUND: Sporadic nodular goiter is a common problem in the United States and significant compressive symptoms may occur with progression to a critical size. METHODS: Potential epidemiological variables associated with the development of large unilateral (> or = 50 g) and bilateral (> or = 100 g) nodular goiter were investigated including: age, gender, race, body mass index (BMI), family history of thyroid disease, pregnancy at time of diagnosis, insurance status, and tobacco or alcohol use. Data were obtained from an IRB-approved thyroid database and retrospective chart review of consecutive patients operated on for nodular goiter from 1990 through 2005. A univariate and multivariate analysis of epidemiological variables in patients with "large" versus "small" nodular goiter was completed. RESULTS: Of the 488 patients operated on for nodular goiter, 113 (23%) were classified as "large," 43 with unilateral (mean 106 +/- 72 g) and 70 with bilateral enlargement (mean 173 +/- 92 g) and 375 (77%) were classified as "small," 179 with unilateral (18 +/- 10 g) and 196 with bilateral (37 +/- 24 g) enlargement. Based on univariate analysis, African-American race, age > or = 40 years, BMI > or = 30 kg/m2, and lack of insurance were associated with an increased risk of large nodular goiter (P < or = 0.001), whereas alcohol use was protective (P = 0.002). A multivariate analysis revealed that African-American race [adjusted odds ratio (adj. OR) 3.3, 95% CI = 2.0-5.4], age > or = 40 years (adj. OR 2.1, 95% CI = 1.2-3.8), and BMI > or = 30 kg/m2 (adj. OR 2.5, 95% CI = 1.5-4.0) were independently associated with large nodular goiter. No significant differences were observed in gender, family history of thyroid disease, pregnancy, or tobacco use (P > 0.1). CONCLUSIONS: African-American race, obesity, and increasing age are independent risk factors for the development of large nodular goiter. These results may be helpful in determining how best to monitor patients with nodular goiter, with earlier intervention to help prevent progressive enlargement and its sequelae. PMID- 16690368 TI - Neutrophil degranulation and the effects of phosphodiesterase inhibition. AB - BACKGROUND: Neutrophils play a major role as the first line in host defense after exposure to bacterial products. However, an exaggerated inflammatory response characterized by overwhelming neutrophil activation can be injurious to the host. Pentoxifylline (PTX), a nonspecific phosphodiesterase inhibitor, has been shown to attenuate neutrophil oxidative burst and decrease proinflammatory mediator synthesis. We hypothesized that PTX down-regulates neutrophil activation by decreasing the surface expression of both CD35 and CD66b, two markers of neutrophil degranulation. MATERIALS AND METHODS: Venous blood was obtained from three healthy volunteers. Whole blood was incubated with HBSS (control), f methionyl-leucyl-phenylalanine (fMLP, 1 microM/L), PTX (2 mM/L), or fMLP + PTX. CD35 and CD66b expression were measured by flow cytometry. RESULTS: fMLP treatment caused a significant increase in CD35 and CD66b expression of when compared to controls (P < 0.01). PTX treatment revealed expression of both markers comparable to the control group. A 38% decrease in CD35 (64 +/- 12 versus 100; P < 0.01) and a 52% decrease in CD66b (48 +/- 7 versus 100; P < 0.01) expression were demonstrated in the fMLP + PTX group when compared to fMLP alone. CONCLUSION: In addition to the known effects of PTX on neutrophil oxidative burst, PTX also affects neutrophil degranulation, an essential step in enzyme release and subsequent tissue injury. These findings may have clinical relevance in the treatment of disease processes due to inflammation in which primed neutrophils play a role. PMID- 16690369 TI - Menin localizes to chromatin through an ATR-CHK1 mediated pathway after UV induced DNA damage. AB - BACKGROUND: Menin is the tumor suppressor protein product of the gene identified in MEN1 syndrome. Evidence suggests menin binds DNA and interacts with proteins implicated in DNA damage pathways. The canonical cellular response to UV-induced DNA damage involves activation of the ataxia-telangiectasia-mutated and Rad3 related (ATR) kinase pathway. MATERIALS AND METHODS: HEK293 cells were irradiated in a UV chamber. Menin's cellular location before and after UV irradiation was investigated by extracting four separate cellular components--a soluble, two chromatin and a nuclear matrix. To block the ATR pathway, we treated with 5 microM of caffeine for 1 h before irradiation. The ATR pathway was further investigated by transiently transfecting HEK293 cells with two mammalian CHK1 expression constructs--full length CHK1 and truncated active CHK1. RESULTS: A 24 h post UV-irradiation time course was studied and demonstrated menin concentration in the chromatin peaked at 4 h. At 4 h post-irradiation, menin concentration in the chromatin increased in a dose dependent manner and demonstrated a 2.8-fold maximal increase. HEK293 cells were pretreated with caffeine, an inhibitor of the ATR. Caffeine decreased menin localization to the chromatin after UV. Constitutively active CHK1 (1-365) transfection increased chromatin-bound menin, mimicking UV irradiation. CONCLUSIONS: Menin localizes to the chromatin after UV irradiation. Caffeine blocks menin localization to the chromatin after UV-irradiation. Over expressing active CHK1 (1-365) increased chromatin-bound menin, similar to UV. The data suggest menin localization to chromatin after UV irradiation is the result of an ATR-CHK1 dependent pathway. PMID- 16690370 TI - Inhibitor development in patients with hemophilia: an overview. PMID- 16690371 TI - Inhibitor economics. AB - The majority of direct costs associated with caring for patients with hemophilia are attributed to replacement therapy with clotting factor concentrates (CFC). For patients who develop high-titer inhibitors, CFC are ineffective and bypassing therapy is used to achieve hemostasis, thus changing the direct costs associated with treatment. As bypassing agents are less predictably effective than CFC (often necessitating more frequent dosing) and are more costly on a per-unit basis, treatment costs for patients with inhibitors are usually much higher than those for patients without inhibitors. In addition, the immune tolerance induction protocols used to eradicate inhibitors are costly due to the frequent dosing of CFC over prolonged periods. It is estimated that the cost of hemostatic therapy for patients with inhibitors can be 2.5 times higher than the cost for patients without inhibitors. However, some studies have reported an outlier effect caused by a small percentage of inhibitor patients who require a disproportionate amount of treatment. These outliers magnify overall treatment costs, making cost assessments for hemostatic therapy less predictable in patients with inhibitors than in those without inhibitors. PMID- 16690372 TI - Surgery in patients with hemophilia and inhibitors: a review of the Norwegian experience with FEIBA. AB - Surgical interventions in patients with hemophilia and inhibitors have often been postponed as long as possible due to difficulties in maintaining intra- and postoperative hemostatic control. Nonactivated and activated prothrombin complex concentrates have been successful in controlling acute bleeding in patients with inhibitors and have been useful in the surgical setting. At the Rikshospitalet Radiumhospitalet University Hospital in Oslo, Norway, 17 minor and seven major surgical procedures were performed in nine patients with congenital hemophilia A and two patients with acquired hemophilia. Patients are generally treated according to the following dosing regimen, with changes made on a case-by-case basis: a preoperative loading dose of 100 U/kg of Factor Eight Inhibitor Bypassing Activity, Anti-Inhibitor Coagulant Complex, Vapor Heated (FEIBA; Baxter AG, Vienna, Austria), followed by 200 U/kg per day for 3 days. The dose is then tapered to 150 U/kg per day and subsequently to 100 U/kg per day. Hemostatic control was attained in all cases and only 1 major adverse event was observed. A 69-year-old patient experienced a non-ST-elevation myocardial infarction 3 days after undergoing a sigmoidectomy. He continued on FEIBA therapy and was stabilized with nitrates, opioid analgesia, and diuretics without clinical signs of heart failure. PMID- 16690373 TI - Overview of inhibitors. AB - Inhibitory antibodies that develop in patients with hemophilia render standard therapy with factor concentrates ineffective. Several factors may influence inhibitor incidence, including genetics, the type of factor concentrate, and environment. A higher incidence of inhibitors in siblings compared to extended relatives, and in African Americans compared to Caucasians, suggests that genetics may play an important role in inhibitor development; however, genetic markers that indicate a predisposition for inhibitor development have yet to be identified. In addition, the appearance of inhibitors in immunologically challenged patients points to the role of the immune response system in the development of inhibitor antibodies, an area that warrants further study. Thus, the medical community faces the difficult task of developing new, improved therapies to combat inhibitors in patients with hemophilia, a task that will require careful consideration of the roles of environmental factors, the immune system, and genetics in inhibitor development. PMID- 16690374 TI - Current opinion on inhibitor treatment options. AB - Inhibitor development in patients with hemophilia complicates hemostatic management. Currently, high doses of factor concentrates are used to override low titer, low-responding inhibitors during acute bleeding episodes. Bypassing agents, such as the activated prothrombin complex concentrate (aPCC), Factor Eight Inhibitor Bypassing Activity, Anti-Inhibitor Coagulant Complex, Vapor Heated (FEIBA; Baxter AG, Vienna, Austria), and activated recombinant factor VII (rFVIIa; NovoSeven, Novo Nordisk A/S, Bagsvaerd, Denmark), are commonly used to treat acute bleeding episodes in patients with high-titer, high-responding inhibitors, whereas long-term therapeutic options include inhibitor eradication using immune tolerance induction. Neither FEIBA nor rFVIIa can be monitored with a laboratory assay, making it difficult to establish optimal dosages. Comparative studies evaluating the efficacy of FEIBA and rFVIIa for bleed control have been sparse, prompting investigators to initiate crossover comparison studies to assess the efficacy and cost of aPCC and rFVIIa in the treatment of joint hemorrhages. Both the cost of therapy and the outcome of therapy will need to be considered in the development of future hemostatic agents for patients with inhibitors. PMID- 16690376 TI - Ablation of atrial fibrillation. AB - Due in large part to the growing dissatisfaction with traditional pharmacologic approaches to atrial fibrillation (AF) management, and fueled by both the fruits of basic and clinical investigation into the nature of AF initiation and maintenance and the explosive development in catheter-based technologies, AF ablation has matured from a purely investigational technique to a viable--and often preferred--strategy for treating patients suffering from this arrhythmia. Future insights and developments will help us refine our treatment strategies, making AF ablation safer, faster, and more effective. As the prevalence of AF in the general population continues to expand, the goal of optimizing our approach to AF becomes ever more important--and more pressing. PMID- 16690377 TI - Modernized Chiropractic reconsidered: beyond foot-on-hose and bones-out-of-place. PMID- 16690379 TI - The effect of chiropractic adjustments on movement time: a pilot study using Fitts Law. AB - OBJECTIVE: To investigate the effect of chiropractic adjustments on movement time using Fitts Law. METHODS: This was a prospective, randomized controlled trial. Ten patients from a private chiropractic practice participated. Participants in the treatment group received high-velocity, low-amplitude chiropractic adjustments to areas of joint dysfunction (chiropractic subluxation). A nonintervention group was used to control for improvement resulting from time and practice effects. Movement time was measured as participants moved a cursor onto a target appearing on a computer screen. A range of target widths and target distances were used to vary the index of difficulty. RESULTS: All participants in the experimental group had significantly improved movement times following spinal adjustments compared with only 1 participant in the control group. The average improvement in movement time for the experimental group was 183 ms, a 9.2% improvement, whereas the average improvement in movement time for the control group was 29 ms, a 1.7% improvement. The difference (improvement) scores after the intervention were significantly greater for the chiropractic group compared with the control group as measured by a 2-tailed independent samples t test (P < .05). CONCLUSION: The results of this study demonstrated a significant improvement in movement time with chiropractic care. These results suggest that spinal adjustments may influence motor behavior. PMID- 16690380 TI - Effect of chiropractic care on heart rate variability and pain in a multisite clinical study. AB - OBJECTIVE: The purpose of this study is to investigate the effect of chiropractic care in a multiclinic setting on sympathetic and parasympathetic nervous system activities using heart rate variability (HRV) analysis. METHODS: Physicians of chiropractic in private practice were provided with an HRV device to perform analysis before and after chiropractic adjustments on 10 subjects. At each site, 8 subjects were monitored before and after a single chiropractic adjustment, and 2 additional patients were followed for a 4-week period with 2 HRV recordings per week. Patient information forms and a visual analog scale (VAS) questionnaire were completed both before and after each chiropractic adjustment. RESULTS: Data from 96 physicians were divided into single-visit and 4-week groups. After 1 chiropractic adjustment, pain as analyzed by VAS was reduced significantly from 3.7 +/- 2.2 to 2.1 +/- 2.0 (P < .001). The mean heart rate reduced from 76.7 +/- 12.7 to 74.3 +/- 12.4 (P < .01), the SD of normal-to-normal QRS increased from a range of 55.8 to 44.6 to a range of 60.6 to 47.2 (P < .001), the high-frequency component increased from 359 +/- 968 to 444 +/- 1069 (P < .01), the low-frequency component increased from 403 +/- 753 to 465 +/- 755 (P < .05), and the total power increased from 1063 +/- 1886 to 1265 +/- 2048 (P < .01). After 4 weeks of chiropractic adjustments, pain measured by the VAS was reduced significantly before and after each visit as analyzed by t tests, but the significant changes were not found using analysis of variance analysis. The reduction of pain from each treatment was not maintained over the 4 weeks of study period. The analysis of variance on the HRV 4-week data found that changes in the SD of normal-to normal QRS, total power, and low-frequency components reached statistically significant levels (P < .05). The heart rate and the high-frequency component did not change significantly (P > .05). CONCLUSION: In this study, HRV and VAS changed in patients as a result of chiropractic care. PMID- 16690381 TI - Fatigue in the sternocleidomastoid muscle and hip dysplasia: a surface electromyographic assessment in adult women. AB - OBJECTIVE: The aim of this study is to assess a possible relationship between unilateral hip dysplasia and sternocleidomastoid muscle function during a fatiguing task. METHODS: In this patient-control, cross-sectional study, 11 female patients (aged 37-63 years) with unilateral coxoarthrosis subsequent to hip dysplasia and 9 female control subjects matched for age were assessed. Surface electromyography of the right and left sternocleidomastoid muscles was measured in the women performing a standardized endurance test. The median power frequency was calculated at the beginning (T0) and at the end of the task (T1, endurance time). For each subject and muscle, percentage decrements in the median power frequency were computed at T1. RESULTS: Endurance time ranged between 15 and 125 s (control group), and 10 and 200 s (patient group), without a significant difference between the 2 groups (Mann-Whitney test, P > .05). In the patient group, no significant differences in the median power frequency between the ispilateral to coxoarthrosis and contralateral sides were found (Wilcoxon test, P > .05). The percentage decrements of the median power frequency in the control (right and left side pooled) and patient group did not significantly differ either in the ispilateral to coxoarthrosis and contralateral sides (Mann Whitney test, P > .05). CONCLUSIONS: The lack of significant differences between the sternocleidomastoid muscles of healthy women and those of women with hip dysplasia does not support the presence of a common noxa causing both congenital muscular torticollis and hip joint alterations. Alternatively, the coexistence of the 2 alterations in infancy may disappear during adult life. PMID- 16690382 TI - A nonsurgical approach to the management of patients with cervical radiculopathy: a prospective observational cohort study. AB - OBJECTIVE: The objective of this study was to describe the clinical outcomes of patients with cervical radiculopathy (CR) treated nonsurgically after a diagnosis based decision rule. METHODS: A prospective observational cohort study on consecutive patients with CR was performed. Data on 35 consecutive patients were collected at baseline, at the end of the active treatment, and at a minimum of 3 months after cessation of treatment. Disability was measured using the Bournemouth Disability Questionnaire. Pain intensity was measured using the Numerical Pain Rating Scale. Patients were also asked to self-rate their improvement. RESULTS: Complete outcome data were available for 31 of the 35 patients. Twenty-seven patients were reached for long-term follow-up. The mean number of months from last treatment to follow-up was 8.2 months. Seventeen patients (49%) reported their improvement as "excellent" and another 14 (40%) did so as "good." The mean patient-rated improvement was 88.2%. The mean percentage of improvement in the Bournemouth Disability Questionnaire score was 78%. The mean percentage of improvement in the Numerical Pain Rating Scale score was 72%. Twenty-four of 31 (77.4%) patients had a clinically significant improvement from baseline to the end of treatment, and 25 of 27 (92.6%) had a clinically significant improvement from baseline to long-term follow-up. CONCLUSIONS: The management strategy that we studied yielded favorable outcomes in this patient sample and appears to be a safe option for patients with CR. However, the absence of randomization and a control group limits interpretation with regard to clinical effectiveness. Randomized clinical trials are necessary to distinguish treatment effects from the natural history of CR. PMID- 16690383 TI - Symptomatic outcomes and perceived satisfaction levels of chiropractic patients with a primary diagnosis involving acute neck pain. AB - OBJECTIVE: The aim of this study was to determine the extent to which a group of patients with acute neck pain managed with chiropractic manipulative therapy benefited from chiropractic care and the degree to which they were subsequently satisfied. METHODS: A two-part retrospective survey, each composed of 14 questions. One part was completed by practicing doctors of chiropractic concerning various aspects of their treatment for patients with former acute neck pain. In the second part, these same patients responded to a telephone survey to measure pre- and posttreatment pain levels and their level of satisfaction with the treatment they received. Chiropractic manipulative therapy was the primary independent variable, although other therapies were used, such as physical therapy, nutritional advice, and exercise. RESULTS: A total of 115 patients were contacted, of whom 94 became study participants, resulting in 60 women (64%) and 34 men. The mean age was 39.6 years (SD, 15.7). The mean number of visits was 24.5 (SD, 21.2). Pain levels improved significantly from a mean of 7.6 (median, 8.0) before treatment to 1.9 (median, 2.0) after treatment (P < .0001). The overall patient satisfaction rate was 94%. CONCLUSION: Patients with acute neck pain involved in this study seemed to be satisfied with chiropractic treatment and reported reductions in associated pain levels and activity restrictions. However, because of the study's design and limitations, care must be taken before drawing firm conclusions from the data presented. PMID- 16690384 TI - Development of a hand dynamometer for the control of manually applied forces. AB - OBJECTIVE: The aim of the study was to develop a prototype tool capable of providing instantaneous feedback about manually applied forces, and to use it to determine (1) whether force constancy can be maintained during different application postures and (2) whether sensitivity to stiffness using the tool is different from sensitivity when assessing stiffness with the hands. METHODS: Subjects were students and staff in a health sciences faculty. A series of grade II and IV mobilizing forces were delivered using a mobilizing tool, a modified JAMAR dynamometer fitted with a molded handle. Forces were executed without feedback at application points on a plinth that were near, midway, and far from the body. Thereafter, discrimination between elastic stiffness was determined with hands alone or using the tool. RESULTS: Force was maintained at grade II when the point of application was moved away from the body, but attempts to replicate grade IV mobilization forces in similar postures showed a decrease despite increased effort. Variation in force produced was substantial both within and between subjects. Stiffness discrimination was not significantly different whether hands or the tool was used. CONCLUSION: There is substantial variation in manually applied forces that could be controlled if instantaneous force readout was available. Reports of hand injury in manual therapists motivate further development of devices that maintain sensitivity and allow for control of applied force and spread the load over a greater surface area on the hands. PMID- 16690385 TI - Electromyogram and force patterns in variably timed manual muscle testing of the middle deltoid muscle. AB - OBJECTIVE: The objective of the study was to compare force curves and surface electromyogram from examiners and subjects during manual muscle testing with 3 examiner-identified variants of manual muscle testing (MMT)-examiner-started (ESMMT), patient-started (PSMMT), and undifferentiated/near-simultaneous (NSMMT). METHODS: Forty-two volunteer applied kinesiologist doctors tested 36 volunteer students, doctors, and spouses at a professional conference. Start-time difference between examiner and subject muscle contraction, peak force, time of peak force and duration of force was measured. Force and surface electromyogram from examiner and subject were recorded simultaneously during MMT of the middle deltoid muscle for each style of testing used in practice. RESULTS: The congruence between examiner label and timing was as follows: ESMMT, 39%; PSMMT 61%; and NSMMT 28% within 50 milliseconds of simultaneous. Mean subject/examiner start-time difference differed significantly between PSMMT (-0.116 seconds) and both ESMMT (-0.018 seconds) and NSMMT (-0.0053 seconds). No clear cutoff between styles was evident. Peak force ranged from 0.548 to 23.615 lb (mean, 8.806 lb; SD, 4.543 lb), and all styles were significantly different. Duration of force ranged from 0.325 to 3.490 seconds (mean, 1.338 seconds; SD, 0.576 seconds), with NSMMT significantly shorter than ESMMT or PSMMT. The shape of force curves did not differ between styles of muscle testing, but differed between facilitated vs inhibited tests. CONCLUSIONS: In this group of doctors, neither start-time difference nor the shape of force curves distinguished styles of MMT. Differences in peak force and test duration may account for differences in outcomes when examiners purposely vary their muscle testing style. PMID- 16690386 TI - Teaching diagnostic decision making: student evaluation of a diagnosis unit. AB - OBJECTIVE: The aim of the study was to report on chiropractic students' response to a changed learning format and on how the students fared in their final examination. METHODS: A 13-week year-4 diagnosis unit that used a structured self study guide, a prescribed text, classroom clinical simulations, WebCT-based learning, and self-assessment tasks is described. After 4 weeks of study, a questionnaire was used to assess the chiropractic students' interim evaluation of the various learning experiences. The ability of students to meet unit objectives was assessed in a summative examination. RESULTS: Most students found the learning experiences acceptable and believed the learning options provided would enable them to meet the unit aims. This belief was confirmed both on formative and summative student assessment. CONCLUSION: Diverse classroom teaching/learning opportunities combined with self-learning guides provide a viable alternative to more traditional teaching formats for introducing chiropractic students to differential diagnosis. PMID- 16690387 TI - Manual forces applied during posterior-to-anterior spinal mobilization: a review of the evidence. AB - OBJECTIVE: The objective of this review was to evaluate the evidence for the consistency of force application by manual therapists when carrying out posterior to-anterior (PA) mobilization techniques, including the factors that influence the application and measurement of mobilization forces. METHODS: Studies were identified by searching 6 electronic databases up to April 2005, screening the reference lists of retrieved articles, and contacting experts by e-mail. Relevant articles were defined as those that described the measurement of forces applied during spinal mobilization or discussed the reliability of measurement of manual forces. RESULTS: Twenty studies described the quantitative measurement of applied force during a PA mobilization technique, with most focusing on the lumbar spine. When defined by magnitude, frequency, amplitude, and displacement, PA mobilization forces are extremely variable among clinicians applying the same manual technique. Variability may be attributed to differences in techniques, measurement or reporting procedures, or variations between therapists or between patients. CONCLUSIONS: The inconsistency in manual force application during PA spinal mobilization in existing studies suggests that further studies are needed to improve the clinical standardization of manual force application. Future research on mobilization should include forces applied to the cervical and thoracic spines in addition to the lumbar spine while thoroughly describing force parameters and measurement methods to facilitate comparison between studies. PMID- 16690388 TI - Cerebrovascular accident without chiropractic manipulation: a case report. AB - OBJECTIVE: To discuss the case of a patient with chronic headache. Although not in severe pain at time of consultation, signs and symptoms raised concern. The patient later had a cerebrovascular accident. CLINICAL FEATURES: A 49-year-old man with non-traumatic chronic episodic head and neck pain presented for care. Examination and plain film radiographs were unremarkable, suggesting a mechanical origin for the symptoms; however, information in the case history raised concerns. INTERVENTION AND OUTCOME: The patient was examined and not manipulated by the doctor of chiropractic but referred back to his general practitioner for a second opinion. The following week, the patient was admitted to hospital having had a cerebrovascular accident. CONCLUSION: The possible indication of the prodrome to a stroke may lie in the case history rather than the examination findings and provocative testing. PMID- 16690389 TI - An etiologic model to help explain the pathogenesis of cervical artery dissection: implications for cervical manipulation. AB - We present a theoretical etiologic model of cervical artery dissection for the purpose of coalescing our present understanding of its pathogenesis. Although the notion of an underlying arteriopathy and a trigger are universally accepted concepts, we propose in our model that dissection is a product of an underlying (genetic) predisposition, triggered specifically by risk factors associated with environmental exposure, with or without trivial trauma. Given the widespread daily occurrence of neck movements and sporting activities, it seems unlikely that trivial trauma, in the absence of other triggers and an underlying arteriopathy, is sufficient to cause dissection. This concept has significant implications for practitioners of spinal manipulation. This may represent a paradigm shift because the model suggests that stroke following manipulation is unlikely in otherwise healthy individuals. Although this model is consistent with current published case-control studies on cervical artery dissection, further research is necessary to accept or refute it as a tenable hypothesis. PMID- 16690390 TI - Targets in ALS: designing multidrug therapies. AB - Amyotrophic lateral sclerosis (ALS) is an incurable disease that arises from the progressive loss of motoneurons. Even when caused by a single gene defect, as in the case of mutations in the enzyme Cu-Zn superoxide dismutase (SOD1), ALS is the result of a complex cascade that involves crosstalk among motoneurons, glia and muscles, and evolves through the action of converging toxic mechanisms. Transgenic rodents that express human mutant SOD1 and develop a progressive paralytic disease are widely used to screen potential therapeutics. Treatments that interfere with a specific event in the neurotoxic cascade have been reported to produce a modest increase in rodent lifespan. Multi-intervention approaches, including novel methods to intercept the damage and to deliver molecules to vulnerable cells, have recently been shown to be more effective. Thus, new avenues for promising therapeutic approaches can be derived from multidrug treatments and/or the delivery of growth factors by viral vectors, in combination with exercise and/or diet regimens. PMID- 16690392 TI - Waiting to exhale. PMID- 16690393 TI - Economies of scale vs. quality of care. PMID- 16690395 TI - Watching for warning signs. PMID- 16690398 TI - Mythbusting volunteerism & experience. PMID- 16690399 TI - Managing those youngsters. PMID- 16690400 TI - Cuss bucket. Paying your insurance. PMID- 16690402 TI - The HAINES position. Australia's answer for first response spinal immobilization has arrived. PMID- 16690403 TI - Patient refusal. What to do when medical treatment & transport are rejected. PMID- 16690404 TI - How Paul Phrampus, MD, got hooked on EMS. PMID- 16690405 TI - Double-click & megabite this! PMID- 16690406 TI - Immunoregulation of autoimmunity by natural killer T cells. AB - Natural killer T (NKT) cells are a conserved subpopulation of lymphocytes that recognize glycolipid antigens in a CD1d context. Upon activation through their semi-invariant T cell receptor, these cells rapidly release large amounts of immunomodulating Th1 and Th2 cytokines. NKT cells have therefore been implicated in immune responses controlling various diseases, including infection, cancer, transplantation, and autoimmunity. Stimulation of the immunoregulatory capacity of NKT cells by the prototypical antigen alpha-galactosylceramide results in amelioration of disease in several animal models. This review will focus on the current knowledge of human NKT cells and their role in autoimmune diseases. The features of these cells and their importance in regulation of autoimmunity suggest that NKT cell-based therapies might be an interesting approach for the treatment of autoimmune diseases. PMID- 16690407 TI - No in vitro evidence for a decreased alloreactivity toward noninherited maternal HLA antigens in healthy individuals. AB - Pre- and/or perinatal exposure to noninherited maternal HLA antigens (NIMA) is associated with a decreased HLA antibody formation against the NIMA and a significantly better graft survival of kidney grafts from siblings or those from unrelated donors who were mismatched for the NIMA haplotype compared with the NIPA (noninherited paternal HLA antigens) haplotype later in life. These observations suggest that some form of immunological tolerance against NIMA is induced. We analyzed the in vitro T cell reactivity of healthy individuals toward their parents and/or siblings expressing the NIMA or NIPA haplotype to explore whether the alloimmune response to NIMA has distinct characteristics compared with NIPA. No differences were detected by mixed lymphocyte reactions (MLR) and supernatants taken from the MLR showed no differences in IFN-gamma and IL-10 production. Additionally, no differences were found with IFN-gamma and IL-10 Elispot analyses. Phenotypic analysis revealed no selective increase in the number of CD3-CD8dim cells (thought to be a NK-like regulator cell) and the number of CD4+CD25+CD152+ cells (naturally occurring regulatory T cells) after stimulation with NIMA-expressing cells when compared with NIPA-expressing cells. In conclusion, no evidence of an influence of a NIMA effect on the cellular level was found in healthy individuals with "standard" immunological techniques. PMID- 16690408 TI - Pemphigus vulgaris is associated with the transporter associated with antigen processing (TAP) system. AB - Pemphigus vulgaris (PV) is a human leukocute antigen (HLA) class II-associated autoimmune disease of the skin of unknown etiology. We recently described the association of pemphigus vulgaris with two clusters of microsatellite loci within the major histocompatibility complex region. One cluster includes the microsatellite marker TAP1CA, located in proximity to the transporter associated with antigen processing (TAP) genes. These genes are essential for class I antigen processing machinery and could be an additional set of genes involved in susceptibility to PV. The aim of this study was to investigate a possible association between TAP gene polymorphisms and PV. For this purpose we examined 37 unrelated Jewish Israeli patients with PV and compared them with 37 healthy Israeli Jewish HLA-matched controls. Significant differences were detected in TAP2 amino acid residues (p=0.001). Two PV TAP2 risk alleles were identified (TAP2*C and TAP2*D), the frequency of which was estimated to be 37.8% in the patients and 5.3 % in the controls. This association was found to be independent of HLA-DR. It is therefore likely that TAP2 genes are involved in susceptibility to development of PV. PMID- 16690409 TI - NKG2A and CD56 are coexpressed on activated TH2 but not TH1 lymphocytes. AB - NKG2A is commonly expressed on cytotoxic cells but has been found on activated T helper (TH) cells. In identifying novel markers differentiating between TH1 and TH2 lymphocytes, we focused on NKG2A expression. TH1 and TH2 cells were negatively isolated from healthy volunteers for microarray analysis and reverse transcription polymerase chain reaction (RT-PCR). Flow cytometry of quiescent and activated TH1 and TH2 cells was performed. Isolates were >95% pure CD3+CD4+ cells (TH1=90.3% and TH2=84.1%). Microarrays revealed differential expression of NKG2A and NKG2C isoforms between TH1 and TH2 cells. RT-PCR indicated greater expression of NKG2A in TH2 cells (4-fold) and NKG2C in TH1 cells (3-fold). Flow studies revealed tripling of TH2 NKG2A with activation to 10.76+/-4.01% (p=0.05), a 23 fold increase in CD56 to 35+/-14.54% (p=0.03), and an increase in NKG2A+CD56+ double-positive cells to 3.04+/-1.38% (p=0.04). TH1 lymphocytes did not differ with activation. We identified co-induction of NKG2A and CD56 on activation of TH2 cells. These cells would likely bind more HLA-E and exhibit increased effector inhibition. Given that certain viruses are known to decrease MHC class I and thus HLA-E production by antigen-presenting cells, activated TH2 cells would bind less HLA-E in this scenario. This would likely result in less effector inhibition and a relatively robust TH2 response. PMID- 16690410 TI - Analysis of a functional BTNL2 polymorphism in type 1 diabetes, rheumatoid arthritis, and systemic lupus erythematosus. AB - The aim of this study was to test whether the functional variant rs2076530 of the BTNL2 gene confers susceptibility to the autoimmune diseases type 1 diabetes (T1D), rheumatoid arthritis (RA), and systemic lupus erythematosus (SLE). Our study populations consisted of 326 patients with T1D and 351 healthy subjects, 808 patients with RA and 1137 healthy controls, and 372 patients with SLE and 280 healthy controls. Genotyping of the BTNL2 gene rs2076530 polymorphism was performed by real-time polymerase chain reaction technology, using the TaqMan 5' allele discrimination assay. We observed statistically significant differences in the distribution of BTNL2rs2076530 alleles between patients with T1D, RA, and SLE and healthy controls (p=0.0035, 0.000003, and 0.00002, respectively), but in two divergent ways: the G allele was associated with T1D and RA, and the A allele was associated with SLE. However, the polymorphism exhibited strong linkage disequilibrium with HLA DQB1-DRB1 haplotypes previously identified as predisposing to the diseases. When the BTNL2 polymorphism was tested conditional on HLA DQB1-DRB1haplotypes, the BTNL2 effect was no longer significant in all three study populations. The BTNL2 rs2076530 polymorphism is associated with T1D, RA, and SLE because of its strong linkage disequalibrium with predisposing HLA DQB1-DRB1 haplotypes in Caucasian populations. PMID- 16690411 TI - Genetic influence of PTPN22 R620W polymorphism in tuberculosis. AB - The PTPN22 gene codes for an intracellular lymphoid-specific phosphatase (Lyp) that has a negative regulatory effect on T-cell activation. Because Lyp is an important molecule involved in the inflammatory response, and its levels are increased in cells that participate in the immune response against Mycobacterium tuberculosis, we hypothesized that the functional PTPN22 C1858T polymorphism could be a genetic factor predisposing to the development of tuberculosis (TB). Accordingly, we undertook an association study in which 113 patients with pulmonary TB and 161 matched healthy controls stratified by the tuberculin skin test (TST) were examined. Significant skewing was observed when T allele frequencies of patients with TB and all controls were compared (P=0.04, odds ratio=0.3; 95% confidence interval=0.08-1.04) and frequencies of patients with TB and TST+ healthy controls were compared (P=0.01, odds ratio=0.2; 95% confidence interval=0.05-0.79). No stratification was detected between patients and control samples. These results suggest that the T allele may be a factor protecting against development of TB once the immune system recognizes M. tuberculosis (i.e., TST+ individuals), whereas the C allele may be a risk factor for development of overt TB. The results also indicate that an association opposite that between the PTPN22 polymorphism and TB exists between TB and autoimmunity. PMID- 16690412 TI - Identification of two novel DQA1 alleles, DQA1*0107 and DQA1*0602, by sequence based typing in the GoKinD population. AB - Two novel DQA1 alleles, DQA1*0107 and DQA1*0602, were discovered using DQA1 sequence-based typing (SBT) in participants in the Genetics of Kidneys in Diabetes (GoKinD) Study. The DQA1*0107 allele, found in three unrelated Caucasian participants, contains a novel polymorphism at codon 79 of exon 2 (CGC-->TGC), which results in an amino acid change from an arginine to a cysteine. The participants containing this novel polymorphism also had a 1-bp insertion in intron 2 that is common to the *01 alleles. The DQA1*0602 allele, found in one Caucasian participant, contains a novel polymorphism at codon 139 of exon 3 (AGC- >CGC), which results in an amino acid change from a serine to an arginine. Additionally, the *0602 allele has a base change in intron 1 that is common to the *06 alleles. Both new alleles were isolated using single-allele amplification SBT and confirmed using sequence-specific primer amplification. PMID- 16690413 TI - The replacement mutation in HLA-DRB1*1211 affects a likely keystone position. AB - Currently, 10 different amino acid variants of the HLA-DRB1*12 family are known. We here report the identification of a new HLA-DRB1*12 allele in a healthy Caucasian male individual. The allele was detected by sequencing-based typing during confirmatory high-resolution typing of an unrelated, male, potential donor from the Czech National Marrow Donors Registry. Compared with DRB1*120101, to which it is closest, the new variant is characterized by a new replacement mutation (T-->C) at nucleotide position 126 of exon 2, resulting in the amino acid substitution Phe-->Leu at position 47. Computational analysis reveals that position 47 functions as a keystone in the beta(1) domain, joining both segments of the alpha helix with the beta sheet, and plays a major role in the structural conformation of the binding groove. Additionally, position 47 is part of pocket E of the peptide binding groove and is directly involved in peptide binding. The new allele, DRB1*1211, is therefore likely to differ substantially from other DRB1*12 alleles in its peptide binding repertoire and alloreactive potential. PMID- 16690414 TI - Association of interleukin-10 A-592C and T-819C polymorphisms with type 2 diabetes mellitus. AB - Type 2 diabetes mellitus (T2DM) is the most common form of diabetes. The association of low interleukin (IL)-10 production capacity with the metabolic syndrome and T2DM was recently established. Approximately 75% of the variation in IL-10 secretion capacity in humans derives from genetic factors that contribute to disease susceptibility. Based on the facts that IL-10 secretion ability is tightly controlled at the transcription level and the low production capacity of IL-10 is associated with T2DM, it seemed tempting to investigate if polymorphisms in the IL-10 gene promoter contribute to T2DM. IL-10 promoter polymorphisms at positions -592 and -819 among 370 consecutive patients with T2DM seen at the Department of Internal Medicine, Chung Shan Medical University Hospital, were examined using polymerase chain reaction restriction fragment length polymorphism. Though no significant association was detected between either the A 592C (p=0.088) or T-819C (p=0.160) polymorphism and T2DM, significantly more T2DM subjects carried -592*C (34.28%, p=0.027) and -819*C (32.57%, p<0.001) alleles, which were associated with high levels of IL-10 production. Nevertheless, no association was observed between these two polymorphisms and biochemical markers for T2DM. Our study suggests that IL-10 genetic polymorphisms may play a specific role(s) in determining diabetic susceptibility, but do not seem to be important in the clinical manifestations of diabetes. PMID- 16690426 TI - The whirligig of time. PMID- 16690427 TI - Dosimetric comparison of two different three-dimensional conformal external beam accelerated partial breast irradiation techniques. AB - PURPOSE: We compare the dosimetry of two techniques for three-dimensional, conformal, external beam, accelerated partial breast irradiation (3D-CPBI) in the supine position. METHODS AND MATERIALS: Sixteen patients with Stage I breast cancer had PBI treatment plans generated using the multiple, noncoplanar photon field technique and the three-field, mixed-modality technique. Planning target volumes (PTVs; lumpectomy site plus 1.5-2.0 cm margin) and total dose (32 Gy) were held constant to facilitate dosimetric comparisons. Plans were optimized for conformality and PTV coverage. RESULTS: Mixed-modality plans employed fewer fields than multiple, noncoplanar photon field plans (mean 3.2 vs. 4.1). Both techniques provided comparable PTV coverage and in all cases, 95% of the PTV received 90% of the prescribed dose. Volumes of ipsilateral breast receiving greater than 16 Gy were similar; however, the mean volume of ipsilateral breast receiving 8 Gy was significantly lower for mixed-modality plans (58% vs. 66%). No differences in the volumes of ipsilateral lung or heart receiving greater than 5 Gy were observed, however, the mixed-modality technique delivered 2.5 Gy to larger volumes of these organs. CONCLUSIONS: Both techniques for supine position, 3D-CPBI provides excellent normal tissue sparing with adequate PTV coverage. The multiple, noncoplanar photon field technique exposes smaller volumes of ipsilateral lung and heart to low dose radiation at the expense of increased plan complexity and larger irradiated breast volumes. PMID- 16690428 TI - Rectal bleeding after high-dose-rate brachytherapy combined with hypofractionated external-beam radiotherapy for localized prostate cancer: impact of rectal dose in high-dose-rate brachytherapy on occurrence of grade 2 or worse rectal bleeding. AB - PURPOSE: To evaluate the incidence of Grade 2 or worse rectal bleeding after high dose-rate (HDR) brachytherapy combined with hypofractionated external-beam radiotherapy (EBRT), with special emphasis on the relationship between the incidence of rectal bleeding and the rectal dose from HDR brachytherapy. METHODS AND MATERIALS: The records of 100 patients who were treated by HDR brachytherapy combined with EBRT for > or =12 months were analyzed. The fractionation schema for HDR brachytherapy was prospectively changed, and the total radiation dose for EBRT was fixed at 51 Gy. The distribution of the fractionation schema used in the patients was as follows: 5 Gy x 5 in 13 patients; 7 Gy x 3 in 19 patients; and 9 Gy x 2 in 68 patients. RESULTS: Ten patients (10%) developed Grade 2 or worse rectal bleeding. Regarding the correlation with dosimetric factors, no significant differences were found in the average percentage of the entire rectal volume receiving 30%, 50%, 80%, and 90% of the prescribed radiation dose from EBRT between those with bleeding and those without. The average percentage of the entire rectal volume receiving 10%, 30%, 50%, 80%, and 90% of the prescribed radiation dose from HDR brachytherapy in those who developed rectal bleeding was 77.9%, 28.6%, 9.0%, 1.5%, and 0.3%, respectively, and was 69.2%, 22.2%, 6.6%, 0.9%, and 0.4%, respectively, in those without bleeding. The differences in the percentages of the entire rectal volume receiving 10%, 30%, and 50% between those with and without bleeding were statistically significant. CONCLUSIONS: The rectal dose from HDR brachytherapy for patients with prostate cancer may have a significant impact on the incidence of Grade 2 or worse rectal bleeding. PMID- 16690429 TI - Resin 90Y-microsphere brachytherapy for unresectable colorectal liver metastases: modern USA experience. AB - PURPOSE: Salvage therapy for patients with unresectable colorectal liver metastases that were refractory to oxaliplatin and irinotecan was performed via radioactive microspheres. High doses of radiation were delivered to tumors from permanently implanted 90Y microspheres, delivered through the hepatic arterial vessels. METHODS AND MATERIALS: Patients from 7 institutions were selected for treatment after screening-defined vascular access to all the tumors, and imaging confirmed microspheres would be implanted only in the liver tumors. All patients were followed with laboratory and imaging studies at regular intervals until death. Toxicities, both acute and late, were recorded, and actuarial survival determined. RESULTS: A total of 208 patients were treated from April 2002 to April 2005. Median follow-up of the 129 men and 79 women is 13 months (range, 1 42 months). Median survival is 10.5 months for responders but only 4.5 months in nonresponders. No treatment-related procedure deaths or radiation-related venoocclusive liver failures were found. Computed tomography partial response was 35%; positron emission tomography response of 91% and reduction in carcinoembryonic antigen of 70% were achieved. CONCLUSIONS: In this group of heavily pretreated patients, radioactive microspheres produced an encouraging median survival, with acceptable toxicity, and a significant objective response rate, which suggests that further investigation of this approach is warranted. PMID- 16690430 TI - Radiation-induced liver disease in three-dimensional conformal radiation therapy for primary liver carcinoma: the risk factors and hepatic radiation tolerance. AB - PURPOSE: To identify risk factors relevant to radiation-induced liver disease (RILD) and to determine the hepatic tolerance to radiation. METHODS AND MATERIALS: The data of 109 primary liver carcinomas (PLC) treated with hypofractionated three-dimensional conformal radiation therapy (3D-CRT) were analyzed. Seventeen patients were diagnosed with RILD and 13 of 17 died of it. RESULTS: The risk factors for RILD were late T stage, large gross tumor volume, presence of portal vein thrombosis, association with Child-Pugh Grade B cirrhosis, and acute hepatic toxicity. Multivariate analyses demonstrated that the severity of hepatic cirrhosis was a unique independent predictor. For Child Pugh Grade A patients, the hepatic radiation tolerance was as follows: (1) Mean dose to normal liver (MDTNL) of 23 Gy was tolerable. (2) For cumulative dose volume histogram, the tolerable volume percentages would be less than: V5 of 86%, V10 of 68%, V15 of 59%, V20 of 49%, V25 of 35%, V30 of 28%, V35 of 25%, and V40 of 20%. (3) Tolerable MDTNL could be estimated by MDTNL (Gy) = -1.686 + 0.023 * normal liver volume (cm3). CONCLUSION: The predominant risk factor for RILD was the severity of hepatic cirrhosis. The hepatic tolerance to radiation could be estimated by dosimetric parameters. PMID- 16690431 TI - Targeting radioimmunotherapy of hepatocellular carcinoma with iodine (131I) metuximab injection: clinical phase I/II trials. AB - PURPOSE: HAb18G/CD147 is a hepatocellular carcinoma (HCC)-associated antigen. We developed iodine (131I) metuximab injection (Licartin), a novel 131I-labeled HAb18G/CD147-specific monoclonal antibody Fab'2 fragment, and evaluated its safety, pharmacokinetics, and clinical efficacy on HCC in Phase I/II trials. METHODS AND MATERIALS: In a Phase I trial, 28 patients were randomly assigned to receive the injection in 9.25-, 18.5-, 27.75-, or 37-MBq/kg doses by hepatic artery infusion. In a multicenter Phase II trial, 106 patients received the injection (27.75 MBq/kg) on Day 1 of a 28-day cycle. Response rate and survival rate were the endpoints. RESULTS: No life-threatening toxic effects were found. The safe dosage was 27.75 MBq/kg. The blood clearance fitted a biphasic model, and its half-life was 90.56-63.93 h. In the Phase II trial, the injection was found to be targeted and concentrated to tumor tissues. Of the 73 patients completing two cycles, 6 (8.22%) had a partial response, 14 (19.18%) minor response, and 43 (58.90%) stable disease. The 21-month survival rate was 44.54%. The survival rate of progression-free patients was significantly higher than that of patients with progressive disease after either one or two cycles (p < 0.0001 or p = 0.0019). CONCLUSION: Iodine (131I) metuximab injection is safe and active for HCC patients. PMID- 16690432 TI - MRI identified prognostic features of tumors in distal sigmoid, rectosigmoid, and upper rectum: treatment with radiotherapy and chemotherapy. AB - PURPOSE: Neoadjuvant therapy is traditionally reserved for locally advanced mid and low rectal cancers. In tumors above this level, the need for adjuvant treatment is based on poor histopathologic features, but this approach has potential disadvantages. The aim of this study was to determine whether magnetic resonance imaging (MRI) could accurately stage tumors of the distal sigmoid, rectosigmoid, and upper rectum and help direct preoperative treatment. MATERIALS AND METHODS: A total of 75 patients with distal sigmoid, rectosigmoid, and upper rectal tumors were assessed preoperatively by MRI. If tumor extended beyond the planned surgical resection plane, chemoradiotherapy was offered. RESULTS: Of the 75 patients, 57 (76%) underwent primary surgery. Agreement between the MRI prognosis and histopathologic findings was 84% (95% confidence interval [CI], 72.6-92.7%). The other 18 patients underwent neoadjuvant chemoradiotherapy for poor prognostic features with predicted surgical resection margin involvement. The histopathologic examination confirmed tumor downstaging in 9 of the 18 patients who underwent chemoradiotherapy. The 3-year survival rate in the good prognosis group (91%; 95% CI, 77.1-97.3%) was not significantly different from that of the chemoradiotherapy group (81.4%; 95% CI, 52.4-93.6%). The poor prognosis group undergoing primary surgery had significantly worse survival (62.2%; 95% CI, 30.3-82.8%, p < 0.03). CONCLUSION: Our findings indicate that tumors of the distal sigmoid, rectosigmoid, and upper rectum can be staged accurately using high spatial resolution MRI and that those with poor prognostic disease may benefit from preoperative therapy. PMID- 16690433 TI - External-beam radiotherapy as preparative regimen for hepatocyte transplantation after partial hepatectomy. AB - PURPOSE: The transplantation of donor hepatocytes is considered a promising option to correct chronic liver failure through repopulation of the diseased organ. This study describes a novel selective external-beam irradiation technique as a preparative regimen for hepatocyte transplantation. METHODS AND MATERIALS: Livers of dipeptidylpeptidase IV (DPPIV)-deficient rats were preconditioned with external-beam single-dose irradiation (25 Gy) delivered to two thirds of the liver. Four days later, a one-third partial hepatectomy (PH) was performed to resect the untreated liver section, and 15 million wild-type (DPPIV+) hepatocytes were transplanted via the spleen into the recipient livers. The degree of donor cell integration and growth was studied 8 h, 3 days, and 5 and 12 weeks after transplantation. RESULTS: Transplanted hepatocytes integrated rapidly into the irradiated liver and proliferated as clusters, finally repopulating the host liver to approximately 20% hepatocyte mass. After 12 weeks, donor cells and their numerous descendents were fully integrated and expressed functional markers to the same extent as host hepatocytes. CONCLUSIONS: We demonstrate that external beam liver irradiation is sufficient to achieve partial repopulation of the host liver after hepatocyte transplantation, under the additional stimulus of one third PH. The method described has potentially good prospects for its application in a clinically viable form of treatment. PMID- 16690434 TI - The vitamin-like dietary supplement para-aminobenzoic acid enhances the antitumor activity of ionizing radiation. AB - PURPOSE: To determine whether para-aminobenzoic acid (PABA) alters the sensitivity of tumor cells to ionizing radiation in vitro and in vivo. METHODS AND MATERIALS: Cellular proliferation was assessed by WST-1 assays. The effects of PABA and radiation on tumor growth were examined with chick embryo and murine models. Real-time reverse transcriptase-polymerase chain reaction and Western blotting were used to quantify p21CIP1 and CDC25A levels. RESULTS: Para aminobenzoic acid enhanced (by 50%) the growth inhibitory activity of radiation on B16F10 cells, whereas it had no effect on melanocytes. Para-aminobenzoic acid enhanced (50-80%) the antitumor activity of radiation on B16F10 and 4T1 tumors in vivo. The combination of PABA and radiation therapy increased tumor apoptosis. Treatment of tumor cells with PABA increased expression of CDC25A and decreased levels of p21CIP1. CONCLUSIONS: Our findings suggest that PABA might represent a compound capable of enhancing the antitumor activity of ionizing radiation by a mechanism involving altered expression of proteins known to regulate cell cycle arrest. PMID- 16690435 TI - Target localization and real-time tracking using the Calypso 4D localization system in patients with localized prostate cancer. AB - PURPOSE: The Calypso 4D Localization System is being developed to provide accurate, precise, objective, and continuous target localization during radiotherapy. This study involves the first human use of the system, to evaluate the localization accuracy of this technique compared with radiographic localization and to assess its ability to obtain real-time prostate-motion information. METHODS AND MATERIALS: Three transponders were implanted in each of 20 patients. Eleven eligible patients of the 20 patients participated in a study arm that compared radiographic triangulated transponder locations to electromagnetically recorded transponder locations. Transponders were tracked for 8-min periods. RESULTS: The implantations were all successful, with no major complications. Intertransponder distances were largely stable. Comparison of the patient localization on the basis of transponder locations as per the Calypso system with the radiographic transponder localization showed an average (+/-SD) 3D difference of 1.5 +/- 0.9 mm. Upon tracking during 8 min, 2 of the 11 patients showed significant organ motion (>1 cm), with some motion lasting longer that 1 min. CONCLUSION: Calypso transponders can be used as magnetic intraprostatic fiducials. Clinical evaluation of this novel 4D nonionizing electromagnetic localization system with transponders indicates a comparable localization accuracy to isocenter, (within 2 mm) compared with X-ray localization. PMID- 16690436 TI - A gradient feature weighted Minimax algorithm for registration of multiple portal images to 3DCT volumes in prostate radiotherapy. AB - PURPOSE: To develop an accurate, fast, and robust algorithm for registering portal and computed tomographic (CT) images for radiotherapy using a combination of sparse and dense field data that complement each other. METHODS AND MATERIALS: Gradient Feature Weighted Minimax (GFW Minimax) method was developed to register multiple portal images to three-dimensional CT images. Its performance was compared with that of three others: Minimax, Mutual Information, and Gilhuijs' method. Phantom and prostate cancer patient images were used. Effects of registration errors on tumor control probability (TCP) and normal tissue complication probability (NTCP) were investigated as a relative measure. RESULTS: Registration of four portals to CTs resulted in 30% lower error when compared with registration with two portals. Computation time increased by nearly 50%. GFW Minimax performed the best, followed by Gilhuijs' method, the Minimax method, and Mutual Information. CONCLUSIONS: Using four portals instead of two lowered the registration error. Reduced fields of view images with full feature sets gave similar results in shorter times as full fields of view images. In clinical situations where soft tissue targets are of importance, GFW Minimax algorithm was significantly more accurate and robust. With registration errors lower than 1 mm, margins may be scaled down to 4 mm without adversely affecting TCP and NTCP. PMID- 16690437 TI - Abdominal organ motion measured using 4D CT. AB - PURPOSE: To measure respiration-induced abdominal organ motion using four dimensional computed tomography (4D CT) scanning and to examine the organ paths. METHODS AND MATERIALS: During 4D CT scanning, consecutive CT images are acquired of the patient at each couch position. Simultaneously, the patient's respiratory pattern is recorded using an external marker block taped to the patient's abdomen. This pattern is used to retrospectively organize the CT images into multiple three-dimensional images, each representing one breathing phase. These images are analyzed to measure organ motion between each phase. The displacement from end expiration is compared to a displacement limit that represents acceptable dosimetric results (5 mm). RESULTS: The organs measured in 13 patients were the liver, spleen, and left and right kidneys. Their average superior to inferior absolute displacements were 1.3 cm for the liver, 1.3 cm for the spleen, 1.1 cm for the left kidney, and 1.3 cm for the right kidney. Although the organ paths varied among patients, 5 mm of superior to inferior displacement from end expiration resulted in less than 5 mm of displacement in the other directions for 41 of 43 organs measured. CONCLUSIONS: Four-dimensional CT scanning can accurately measure abdominal organ motion throughout respiration. This information may result in greater organ sparing and planning target volume coverage. PMID- 16690438 TI - Beam angle optimization and reduction for intensity-modulated radiation therapy of non-small-cell lung cancers. AB - PURPOSE: To optimize beam angles and reduce the number of beams used for intensity-modulated radiation therapy (IMRT) of non-small-cell lung cancer (NSCLC). METHODS AND MATERIALS: An exhaustive search scheme was used to perform beam angle optimization (BAO) for IMRT of NSCLC. This approach involved intercomparison of all possible beam angle combinations and selection of the best angles based on the scores or costs of the objective functions used in the treatment plan optimization. Ten Stage III NSCLC cases were selected to evaluate the BAO algorithm and dosimetry benefits of IMRT-BAO. IMRT plans using five or seven coplanar beams were optimized and compared with those using nine equal spaced beams. Results of BAO were also compared between plans using different numbers of beams with or without fluence modulation. RESULTS: Each anatomic structure, e.g., tumor or lung, had its own preferred beam angles. Thus, BAO required appropriate balance of competing objective functions. Plans using fewer angles (five or seven beams) could achieve plan quality similar to those using nine equal-spaced beams, however with reduced monitor units and field segments. The number of beams used for the treatment (five vs. seven) and the fluence modulation (open or IMRT beams) did not have a significant impact on the results of the BAO. CONCLUSIONS: Use of fewer beams (e.g., five) for lung IMRT could result in acceptable plan quality but improved treatment efficiency. A multiresolution search scheme could be developed for BAO using fewer and nonmodulated beams to reduce the computation cost of BAO. PMID- 16690439 TI - The potential for dose escalation in lung cancer as a result of systematically reducing margins used to generate planning target volume. AB - PURPOSE: To determine how much the radiation dose to lung tumors could be increased as the margins used to generate planning target volume (PTV) are reduced. METHODS AND MATERIALS: Treatment plans for 18 patients with non-small cell lung carcinoma were retrospectively generated. Dose escalation was performed in two phases: The dose was increased as long as healthy tissue dose-volume constraints did not exceed (1) the values from the treatment plan originally used for the patients and (2) clinically acceptable values. RESULTS: No correlation of dose escalation was observed with tumor location, tumor stage, tumor motion, and tumor volume. An increase in dose was observed for many of the patients with as little as 2-mm uniform reduction in PTV margin, with increases in mean PTV dose exceeding 15 Gy for 5 patients. Sixteen of 18 patients experienced a decrease in mean heart, esophagus, and lung dose when margins were reduced and prescription doses were increased. CONCLUSIONS: Reduced margins allowed an increased dose to the tumors. However, a much larger dose escalation was possible for some patients but not for others, demonstrating that each patient is different, so individual treatment plans must be tailored for maximum tumor coverage and minimum exposure of healthy tissue. PMID- 16690440 TI - A study of the effect of setup errors and organ motion on prostate cancer treatment with IMRT. AB - PURPOSE: To assess the influence of setup errors and organ motion in terms of the probability of tumor control and normal-tissue complications by tumor control probability and normal-tissue complication probability. METHODS AND MATERIALS: Twelve patients were treated for prostate cancer with intensity-modulated radiation therapy. Two orthogonal portal images were taken daily. All patients underwent three computed tomography scans during the 8-week treatment time (i.e., baseline, intermediate, and final). The original treatment plans were re evaluated, taking into account setup errors and organ motion. RESULTS: The mean shifts +/- standard deviation of the whole patient population in the lateral, anterior-posterior, and craniocaudal direction were 1.0 +/- 1.5 mm, 0.9 +/- 2.1 mm, and 1.9 +/- 2.1 mm, respectively. In most of the recalculated dose-volume histograms, the coverage of clinical target volume was granted despite organ motion, whereas the rectal wall histograms were often very different from the planned ones. CONCLUSION: We have studied the impact of prostate and rectum motion, as well as setup errors, on dose-volume histograms. The estimate of these effects may have implications for predictive indications when planning intensity modulated radiation therapy treatments on prostate. PMID- 16690441 TI - Prostate contouring uncertainty in megavoltage computed tomography images acquired with a helical tomotherapy unit during image-guided radiation therapy. AB - PURPOSE: To evaluate the image-guidance capabilities of megavoltage computed tomography (MVCT), this article compares the interobserver and intraobserver contouring uncertainty in kilovoltage computed tomography (KVCT) used for radiotherapy planning with MVCT acquired with helical tomotherapy. METHODS AND MATERIALS: Five prostate-cancer patients were evaluated. Each patient underwent a KVCT and an MVCT study, a total of 10 CT studies. For interobserver variability analysis, four radiation oncologists, one physicist, and two radiation therapists (seven observers in total) contoured the prostate and seminal vesicles (SV) in the 10 studies. The intraobserver variability was assessed by asking all observers to repeat the contouring of 1 patient's KVCT and MVCT studies. Quantitative analysis of contour variations was performed by use of volumes and radial distances. RESULTS: The interobserver and intraobserver contouring uncertainty was larger in MVCT compared with KVCT. Observers consistently segmented larger volumes on MVCT where the ratio of average prostate and SV volumes was 1.1 and 1.2, respectively. On average (interobserver and intraobserver), the local delineation variability, in terms of standard deviations [Deltasigma = radical(sigma2MVCT-sigma2KVCT)], increased by 0.32 cm from KVCT to MVCT. CONCLUSIONS: Although MVCT was inferior to KVCT for prostate delineation, the application of MVCT in prostate radiotherapy remains useful. PMID- 16690442 TI - Dosimetric comparisons of helical tomotherapy treatment plans and step-and-shoot intensity-modulated radiosurgery treatment plans in intracranial stereotactic radiosurgery. AB - PURPOSE: To evaluate dose conformity, dose homogeneity, and dose gradient in helical tomotherapy treatment plans for stereotactic radiosurgery, and compare results with step-and-shoot intensity-modulated radiosurgery (IMRS) treatment plans. METHODS AND MATERIALS: Sixteen patients were selected with a mean tumor size of 14.65 +/- 11.2 cm3. Original step-and-shoot IMRS treatment plans used coplanar fields because of the constraint of the beam stopper. Retrospective step and-shoot IMRS plans were generated using noncoplanar fields. Helical tomotherapy treatment plans were generated using the tomotherapy planning station. Dose conformity index, dose gradient score index, and homogeneity index were used in plan intercomparisons. RESULTS: Noncoplanar IMRS plans increased dose conformity and dose gradient, but not dose homogeneity, compared with coplanar IMRS plans. Tomotherapy plans increased dose conformity and dose gradient, yet increased dose heterogeneity compared with noncoplanar IMRS plans. The average dose conformity index values were 1.53 +/- 0.38, 1.35 +/- 0.15, and 1.26 +/- 0.10 in coplanar IMRS, noncoplanar IMRS, and tomotherapy plans, respectively. The average dose homogeneity index values were 1.15 +/- 0.05, 1.13 +/- 0.04, and 1.18 +/- 0.09 in coplanar IMRS, noncoplanar IMRS, and tomotherapy plans, respectively. The mean dose gradient score index values were 1.37 +/- 19.08, 22.32 +/- 19.20, and 43.28 +/- 13.78 in coplanar IMRS, noncoplanar IMRS, and tomotherapy plans, respectively. The mean treatment time in tomotherapy was 42 +/- 16 min. CONCLUSIONS: We were able to achieve better dose conformity and dose gradient in tomotherapy plans compared with step-and-shoot IMRS plans for intracranial stereotactic radiosurgery. However, tomotherapy treatment time was significantly larger than that in step-and-shoot IMRS. PMID- 16690443 TI - Interfractional dose variation during intensity-modulated radiation therapy for cervical cancer assessed by weekly CT evaluation. AB - PURPOSE: To investigate the interfractional dose variation of a small-bowel displacement system (SBDS)-assisted intensity-modulated radiation therapy (IMRT) for the treatment of cervical cancer. METHODS: Four computed tomography (CT) scans were carried out in 10 patients who received radiotherapy for uterine cervical cancer. The initial CT was taken by use of the SBDS, before the beginning of radiotherapy, and 3 additional CT scans with the SBDS were done in subsequent weeks. IMRT was planned by use of the initial CT, and the subsequent images were fused with the initial CT set. Dose-volume histogram (DVH) changes of the targets (planning target volume [PTV] = clinical target volume [CTV] + 1.5 cm) and of the critical organs were evaluated after obtaining the volumes of each organ on 4 CT sets. RESULTS: No significant differences were found in PTV volumes. Changes on the DVH of the CTVs were not significant, whereas DVH changes of the PTVs at 40% to 100% of the prescription dose level were significant (V(90%); 2nd week: p = 0.0091, 3rd week: p = 0.0029, 4th week: p = 0.0050). The changes in the small-bowel volume included in the treatment field were significant. These were 119.5 cm3 (range, 26.9-251.0 cm3), 126 cm3 (range, 38.3 336 cm3), 161.9 cm3 (range, 37.7-294.6 cm3), and 149.1 cm3 (range, 38.6-277.8 cm3) at the 1st, 2nd, 3rd, and 4th weeks, respectively, and were significantly correlated with the DVH change in the small bowel, which were significant at the 3rd (V80%; p = 0.0230) and 4th (V80%; p = 0.0263) weeks. The bladder-volume change correlated to the large volume change (>20%) of the small-bowel volume. CONCLUSIONS: Significant DVH differences for the small bowel can result because of interfractional position variations, whereas the DVH differences of the CTV were not significant. Strict bladder-filling control and an accurate margin for the PTV, as well as image-guided position verification, are important to achieve the goal of IMRT. PMID- 16690444 TI - The Vienna applicator for combined intracavitary and interstitial brachytherapy of cervical cancer: design, application, treatment planning, and dosimetric results. AB - PURPOSE: To present a combined intracavitary and interstitial dedicated applicator and magnetic resonance imaging (MRI) treatment planning for cervical cancer brachytherapy. METHODS AND MATERIALS: A modified ring applicator allows interstitial needles to be implanted in parallel to the intrauterine tandem. MRI treatment planning based on a standard loading pattern with stepwise dwell weight adaptation and needle loading is performed to achieve optimal dose coverage and sparing of organs at risk. Dose constraints are applied for dose-volume histogram parameters. RESULTS: The use of additional interstitial needles provides prescription dose up to 15 mm lateral to point A. Twenty-two patients with high risk clinical target volumes of mean 44 cm3 were treated with a mean prescribed total dose of 85 Gy (biologically equivalent to 2 Gy fractionation, alpha/beta = 10 Gy) and 93% coverage (V100). The dose to organs at risk was within standard limits for intracavitary brachytherapy alone. CONCLUSIONS: A combined interstitial-intracavitary applicator results in reproducible implants for cervical cancer brachytherapy. MRI-based treatment planning based on a target concept, dose-volume constraints, and limitations for the relative dwell weight allows for an increase in target coverage, treated volume, and total dose without increasing the dose to critical structures. PMID- 16690446 TI - In regards to Arcangeli et al. (Int J Radiat Oncol Biol Phys 2006;64:161-167). PMID- 16690448 TI - The efficacy of FDG-PET in head-and-neck cancer with neck metastasis after definitive radiation treatment: in regard to Yao et al. (Int J Radiat Oncol Biol Phys 2005;63:991-999). PMID- 16690449 TI - Abstracts of the 7th European Congress on Menopause, 3-7 June, 2006, Istanbul, Turkey. PMID- 16690450 TI - Cone-beam imaging and respiratory motion (IGRT)-part II. PMID- 16690451 TI - Overview of image-guided radiation therapy. AB - Radiation therapy has gone through a series of revolutions in the last few decades and it is now possible to produce highly conformal radiation dose distribution by using techniques such as intensity-modulated radiation therapy (IMRT). The improved dose conformity and steep dose gradients have necessitated enhanced patient localization and beam targeting techniques for radiotherapy treatments. Components affecting the reproducibility of target position during and between subsequent fractions of radiation therapy include the displacement of internal organs between fractions and internal organ motion within a fraction. Image-guided radiation therapy (IGRT) uses advanced imaging technology to better define the tumor target and is the key to reducing and ultimately eliminating the uncertainties. The purpose of this article is to summarize recent advancements in IGRT and discussed various practical issues related to the implementation of the new imaging techniques available to radiation oncology community. We introduce various new IGRT concepts and approaches, and hope to provide the reader with a comprehensive understanding of the emerging clinical IGRT technologies. Some important research topics will also be addressed. PMID- 16690452 TI - Dynamic targeting image-guided radiotherapy. AB - Volumetric imaging and planning for 3-dimensional (3D) conformal radiotherapy and intensity-modulated radiotherapy (IMRT) have highlighted the need to the oncology community to better understand the geometric uncertainties inherent in the radiotherapy delivery process, including setup error (interfraction) as well as organ motion during treatment (intrafraction). This has ushered in the development of emerging technologies and clinical processes, collectively referred to as image-guided radiotherapy (IGRT). The goal of IGRT is to provide the tools needed to manage both inter- and intrafraction motion to improve the accuracy of treatment delivery. Like IMRT, IGRT is a process involving all steps in the radiotherapy treatment process, including patient immobilization, computed tomography (CT) simulation, treatment planning, plan verification, patient setup verification and correction, delivery, and quality assurance. The technology and capability of the Dynamic Targeting IGRT system developed by Varian Medical Systems is presented. The core of this system is a Clinac or Trilogy accelerator equipped with a gantry-mounted imaging system known as the On-Board Imager (OBI). This includes a kilovoltage (kV) x-ray source, an amorphous silicon kV digital image detector, and 2 robotic arms that independently position the kV source and imager orthogonal to the treatment beam. A similar robotic arm positions the PortalVision megavoltage (MV) portal digital image detector, allowing both to be used in concert. The system is designed to support a variety of imaging modalities. The following applications and how they fit in the overall clinical process are described: kV and MV planar radiographic imaging for patient repositioning, kV volumetric cone beam CT imaging for patient repositioning, and kV planar fluoroscopic imaging for gating verification. Achieving image-guided motion management throughout the radiation oncology process requires not just a single product, but a suite of integrated products to manipulate all patient data, including images, efficiently and effectively. PMID- 16690453 TI - X-ray volume imaging in image-guided radiotherapy. AB - Treatment simulation has significantly improved the accuracy and precision of radiation therapy delivery. A new generation of therapy systems promises to take the simulation and imaging process to a new level of accuracy; however, this will require changes in the workflow process. We describe the first generation of these devices, review the various imaging options and how they might be used in the clinic to improve treatment outcomes, and suggest several workflow approaches. Workflows discussed include on-line interventional, off-line adaptive, and off-line predictive approaches, with both geometric and dosimetric considerations. These changes will place new knowledge requirements on the medical dosimetrist and will necessitate involvement in the development of new departmental processes. PMID- 16690454 TI - Localizing moving targets and organs using motion-managed CTs. AB - Respiration-induced target and organ motion impacts the radiotherapy strategies of some cancers. Various methods and techniques have been used to investigate motion-related radiotherapy issues, including retrospective 4-dimensional (4D) computed tomography (CT), prospective gated CT, and breath-hold CT scans. This paper reviews these methods and, particularly, the method using retrospective 4D CT scans, which has been developed at our institution. Some motion studies based on retrospective 4D CT images of patients are also examined. These studies have led to reduced planning target volume (PTV) margins for a number of patients, because the respiratory motion was observed to be minimal or gated radiotherapy was used. Respiratory motion managed CTs and, particularly, retrospective 4D CTs are proving to be useful for measuring soft tissue motion, identifying patients who could benefit from gated radiotherapy, and evaluating the effects of respiratory motion during radiotherapy. PMID- 16690455 TI - Technical aspects of image-guided respiration-gated radiation therapy. AB - In this review article, we discuss various technical aspects of image-guided respiration-gated radiation therapy. We first review some basic concepts related to respiratory gating, including gating window, duty cycle, residual motion, internal/external gating, amplitude/phase gating, etc. We then discuss 2 implementations of image-guided respiration-gated treatment, i.e., the Mitsubishi/Hokkaido technique for internal gating and the MGH technique for external gating. Several existing problems related to respiratory gating, namely external gating mode (phase vs. amplitude), imaging dose for internal gating, gated treatment for lung cancer without implanted fiducial makers, as well as gated intensity-modulated radiation therapy issues, are also discussed along with potential solutions. PMID- 16690456 TI - The clinical implementation of respiratory-gated intensity-modulated radiotherapy. AB - The clinical use of respiratory-gated radiotherapy and the application of intensity-modulated radiotherapy (IMRT) are 2 relatively new innovations to the treatment of lung cancer. Respiratory gating can reduce the deleterious effects of intrafraction motion, and IMRT can concurrently increase tumor dose homogeneity and reduce dose to critical structures including the lungs, spinal cord, esophagus, and heart. The aim of this work is to describe the clinical implementation of respiratory-gated IMRT for the treatment of non-small cell lung cancer. Documented clinical procedures were developed to include a tumor motion study, gated CT imaging, IMRT treatment planning, and gated IMRT delivery. Treatment planning procedures for respiratory-gated IMRT including beam arrangements and dose-volume constraints were developed. Quality assurance procedures were designed to quantify both the dosimetric and positional accuracy of respiratory-gated IMRT, including film dosimetry dose measurements and Monte Carlo dose calculations for verification and validation of individual patient treatments. Respiratory-gated IMRT is accepted by both treatment staff and patients. The dosimetric and positional quality assurance test results indicate that respiratory-gated IMRT can be delivered accurately. If carefully implemented, respiratory-gated IMRT is a practical alternative to conventional thoracic radiotherapy. For mobile tumors, respiratory-gated radiotherapy is used as the standard of care at our institution. Due to the increased workload, the choice of IMRT is taken on a case-by-case basis, with approximately half of the non-small cell lung cancer patients receiving respiratory-gated IMRT. We are currently evaluating whether superior tumor coverage and limited normal tissue dosing will lead to improvements in local control and survival in non-small cell lung cancer. PMID- 16690457 TI - Abstracts for the American Association of Clinical Endocrinologists 15th Annual Meeting and Clinical Congress, April 26-30, 2006, Chicago, Illinois, USA. PMID- 16690458 TI - Effects of treatment of hyperthyroidism on glucose homeostasis, insulin secretion, and markers of bone turnover. AB - OBJECTIVE: To study glucose and bone metabolism in hyperthyroidism, assess their changes after treatment, and investigate their interrelationships. METHODS: Thirty patients with hyperthyroidism matched with 32 normal control subjects were studied. After a 10- to 12-hour overnight fast, blood samples were collected for measurement of glucose, insulin, C-peptide, and intact proinsulin levels as well as for measurement of bone markers: serum alkaline phosphatase (ALP), osteocalcin, and procollagen type I C-terminal peptide (PICP) as markers of bone formation and serum C-terminal cross-linked telopeptide of type I collagen (ICTP) as a marker of bone resorption. A 3-hour 75-g oral glucose tolerance test was then performed, with measurement of glucose, insulin, and C-peptide levels every 30 minutes. Patients were studied at baseline and after treatment with an antithyroid drug (carbimazole) for 1 month and 6 months. RESULTS: Pretreatment fasting glucose, insulin, C-peptide, and intact proinsulin levels were significantly higher in patients with hyperthyroidism than in control subjects. During the 3-hour oral glucose tolerance test, the area under the curve of glucose was significantly elevated in the patients, whereas the 3-hour areas under the curve of insulin and C-peptide were not significantly different between patients and control subjects. Fasting glucose, insulin, C-peptide, and intact proinsulin levels decreased significantly to levels similar to those of the control subjects after 1 month of antithyroid therapy and remained so at 6 months. Pretreatment ALP, osteocalcin, PICP, and ICTP were significantly higher in the patients than in the control subjects. After treatment, all markers of bone turnover decreased significantly to levels similar to those of the control subjects at 1 month (except ALP) and 6 months. Within the study group of patients, baseline PICP, osteocalcin, and ICTP demonstrated positive correlation trends with free triiodothyronine and free thyroxine. CONCLUSION: Abnormal glucose metabolism and increased bone turnover are hallmarks of untreated hyperthyroidism. These factors normalize as early as 4 weeks after initiation of antithyroid therapy. Changes in bone markers, particularly those of resorption, are related to the degree of thyroid hyperactivity. PMID- 16690459 TI - Parathyroid lipoadenomas: a rare cause of primary hyperparathyroidism. AB - OBJECTIVE: To review one institution's experience with parathyroid lipoadenomas and to report the associated clinical characteristics. METHODS: We present a case series of parathyroid lipoadenomas. A parathyroid lipoadenoma was defined as a single adenoma with more than 50% fat on histologic examination in conjunction with primary hyperparathyroidism and resolution of hypercalcemia postoperatively. Patients who fulfilled the diagnostic criteria were identified from the surgical pathology files of the Mayo Clinic. RESULTS: Five cases of parathyroid lipoadenomas, including 1 oxyphil lipoadenoma, were identified during the period from 1971 to 2001. The clinical picture of the study subjects resembled that of a typical patient with primary hyperparathyroidism. Parathyroid lipoadenomas were identified in 3 women (60%) and 2 men (40%), and only 1 patient presented with possible hypercalcemia-related symptoms of nephrolithiasis and hip fracture, leading to diagnosis. The mean serum calcium concentration was 11.1 mg/dL. Preoperatively, all study subjects had elevation of serum parathyroid hormone levels. Two of 3 patients (67%) had the tumor identified preoperatively by neck ultrasonography. CONCLUSION: A parathyroid lipoadenoma is a rare cause of primary hyperparathyroidism. The clinical features of this pathologic entity are similar to those of the more common pathologic variants of parathyroid disease associated with primary hyperparathyroidism. PMID- 16690460 TI - Reference range for serum parathyroid hormone. AB - OBJECTIVE: To determine whether the reference range for parathyroid hormone (PTH) should be lowered (from 65 pg/mL to a proposed value of 46 pg/mL) with use of the Allegro radioimmunometric assay. METHODS: We examined the reference range for PTH, adjusted for serum 25-hydroxyvitamin D (25-OHD), in 503 healthy African American and white women, who were 20 to 80 years old. We also analyzed other factors that are thought to influence PTH levels. RESULTS: Univariate predictors of PTH were identified, and a multivariate model was developed with use of the variables and PTH. Serum PTH was significantly higher in black study subjects than in white study subjects (P<0.02). Increasing PTH was also significantly correlated with increasing body mass index, age, and serum creatinine and with decreasing dietary calcium intake and serum 25-OHD levels. A stepwise multiple linear regression analysis yielded the following predictors of PTH: body mass index (R2=9.4%), age (R2=1.0%), and serum 25-OHD (R2=0.8%). In our study population, many PTH values were above the proposed new upper limit of 46 pg/mL. CONCLUSION: The upper limit of the reference range for serum PTH should not be changed. Factors to be considered in analysis of serum PTH values in the upper reference range in patients with normocalcemia include obesity, race, 25-OHD levels, advanced age, serum creatinine, and dietary calcium intake. PMID- 16690461 TI - Glycemic relapse in type 2 diabetes. AB - OBJECTIVE: To characterize the occurrence of glycemic relapse after initial improvement in blood glucose levels and to describe predictors of relapse in patients with type 2 diabetes. METHODS: Occurrence of glycemic relapse was analyzed in 393 consecutive patients with type 2 diabetes after participation in a 3-month intensive outpatient intervention. All patients had hemoglobin A1c (A1C) values (3)7% before the intervention and had achieved adequate glycemic control (nadir A1C<7%) afterward. The median follow-up time was 26.5 months. Relapse was defined as a subsequent increase in A1C to (3)7%. RESULTS: The probability of glycemic relapse was 45% at 1 year after the intervention and was 76% at 3 years. The median time to relapse was 15.2 months. Cox multivariate regression analysis indicated that treatment with insulin was associated with a greater risk of relapse-- hazard ratio=1.5 (95% confidence interval, 1.1 to 2.2), after controlling for the patient's age, sex, race, body mass index, duration of diabetes, weight change during the intervention, and nadir A1C value. Among those patients not treated with insulin at the end of the intervention, a shorter duration of diabetes and weight loss during the intervention period were significantly associated with decreased risk of relapse. CONCLUSION: The majority of study patients with type 2 diabetes who attained satisfactory glycemic control after intensive outpatient intervention had a relapse after the end of the intervention period. Patients receiving insulin therapy were at particular risk of glycemic relapse. Therefore, such patients should receive high priority for continuation of intensive care or for other relapse prevention measures. PMID- 16690462 TI - Role of [18F]fluorodeoxyglucose positron emission tomography in follow-up of differentiated thyroid cancer. AB - OBJECTIVE: To assess the diagnostic utility of [(18)F]flu-orodeoxyglucose positron emission tomography (FDG PET) in the follow-up of patients with differentiated thyroid cancer (DTC). METHODS: In this study, we used strict definitions of presence and absence of the disease and performed all FDG PET scans while the patients were not taking levothyroxine (LT4). We report the results of conventional FDG PET scans obtained during the follow-up of 50 nonselected patients with DTC (34 female and 16 male patients; median age, 40.5 years; range, 18 to 68). All FDG PET scans and measurement of thyroglobulin (Tg) levels were performed while the patients were not taking LT4 (thyrotropin>or=25 microIU/mL). Tg antibodies were negative in all patients. The initial surgical procedure was total thyroidectomy in all cases, and 26 patients underwent additional operations (2 to 4 procedures). Radioactive iodine (131I) therapy was given to 48 patients (median dose, 5,550 MBq). In 42 patients, FDG PET was used for evaluation of Tg-positive (Tg>2 ng/mL in the absence of LT4 therapy), scan negative disease. In 8 patients, Tg was 2 ng/mL without LT4 therapy) for >1 year. Disease was considered absent if Tg was 196.20 (Delta(9)-THC-D(3)), 401.15 > 365.2 (25(OH)D(3)), and 413.15 > 355.20 (25(OH)D(2)). Reference interval study results were compared with current 25(OH)D recommendations. Elution of 25(OH)D(2), 25(OH)D(3), and Delta(9)-THC-D(3) was achieved after 3.0 minutes (total run time, 6.0 minutes). Within- and between-run coefficients of variation were less than 11%. Deming regression of radioimmunoassay and LC-MS/MS methods for total 25(OH)D levels yielded a slope of 0.97 (95% confidence interval, 0.88-1.05) and y intercept of -1.74 ng/mL. Reference intervals were less than recommended levels (D(2), 0.0-12.1; D(3), 5.5-41.4; total vitamin D, 6.0-43.5 ng/mL [0-30, 14-103, 15-109 nmol/L, respectively]) with no statistically significant differences in race, age, or sex. This LC-MS/MS method provides a rapid, accurate, sensitive, and cost-effective alternative to other methods for detection of 25(OH)D(2) and 25(OH)D(3) at nanomolar concentrations. PMID- 16690492 TI - Performance characteristics of seven automated CA 125 assays. AB - Cancer antigen 125 (CA 125) is a high-molecular-mass glycoprotein that is used as a tumor marker to monitor disease progression and response to therapy and in early detection of recurrence after treatment for ovarian cancer. The Access 2 (Beckman Coulter, Brea, CA), ADVIA Centaur (Bayer Diagnostics, Tarrytown, NY), ARCHITECT i2000 (Abbott Diagnostics, Abbott Park, IL), AxSYM (Abbott Diagnostics), Elecsys 2010 (Roche Diagnostics, Indianapolis, IN), IMMULITE 2000 (Diagnostic Products, Los Angeles, CA), and VITROS ECi (Ortho Clinical Diagnostics, Raritan, NJ) assays for CA 125 were evaluated for detection limit, dilution linearity, imprecision, correlation, and reference intervals. The maximum average deviation from target recoveries for dilution linearity studies ranged from 3.7% for the ADVIA Centaur to 18.2% for the IMMULITE 2000. Imprecision studies yielded total coefficients of variation of 2.0% to 8.3% at CA 125 concentrations of 35 and 114 U/mL (35 and 114 kU/L). Method comparison studies revealed good agreement with the VITROS ECi comparison method, with slopes ranging between 0.88 to 1.19 and correlation coefficients of more than 0.95. All methods show acceptable performance characteristics and generally compare well. However, for some samples, substantial differences exist between methods, necessitating parallel testing when introducing a new method. PMID- 16690493 TI - Three-repeat Tau 69 is a major tau isoform in laser-microdissected Pick bodies. AB - By utilizing a novel combinatorial method of a Laser Microdissection System and Western blot analysis, we demonstrate that a distinct isoform of abnormally phosphorylated tau (69 kDa, Tau 69) predominantly aggregated in laser microdissected Pick bodies (PBs) in sporadic Pick's disease. By contrast, tau migrated as two major bands of 60 and 64 kDa (Tau 60 and 64) in total brain homogenates as previously reported. Comparative immunohistochemical analysis with anti-4-repeat antibody revealed that a major component of the abnormally phosphorylated tau in these PBs was 3-repeat tau (3R-tau). Whether 29 amino acid repeat encoded by exons 2 and 3 in the Tau 69 might accelerate the formation of PBs remains to be further investigated. Such a combination of morphological and biochemical techniques significantly complements the existing histopathological methods. PMID- 16690494 TI - Protective effect of Rosa laevigata against amyloid beta peptide-induced oxidative stress. AB - The amyloid beta (Abeta) peptide is known to increase free radical production in nerve cells, leading to cell death. To investigate the effect of Rosa laevigata against Abeta-induced oxidative damage, in vitro assays and in vivo behavioral tests were performed. R. laevigata showed cell protective effects against oxidative stress-induced cytotoxicity. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide) reduction assay exhibited significant increase in cell viability when rat pheochromocytoma (PC 12) cells were treated with R. laevigata extracts. Administration of R. laevigata extracts to mice significantly reversed the Abeta-induced learning and memory impairment in in vivo behavioral tests. These results suggest that R. laevigata extracts can reduce the cytotoxicity of Abeta in PC 12 cells, possibly by the reduction of oxidative stress, and these extracts may be useful in the prevention of Alzheimer's disease. PMID- 16690495 TI - Glycosaminoglycans are part of amyloid fibrils: ultrastructural evidence in avian AA amyloid stained with cuprolinic blue and labeled with immunogold. AB - In domestic brown layer fowl, reactive amyloidosis of internal organs, such as liver and spleen, and of the joints is a common disorder. In a variety of amyloid types including the AA-amyloid of the chicken, in addition to amyloid fibrils, proteoglycans and glycosaminoglycans (GAGs) are found on immunohistochemistry or after extraction. The aim of the present report is to study amyloid fibrils for the ultrastructural location of GAGs by cuprolinic blue staining and immunogold labeling. Rabbit antichicken AA antiserum was used for the immunogold labeling on conventionally embedded and cryoembedded liver tissue and revealed similar results. Therefore conventional blocks could be used for further analysis. Cuprolinic blue staining was performed on blocks of joint tissue in which clearly discernable rod-shaped glycoproteins were encountered in between collagen fibrils. Moreover, it appeared to stain larger deposits which might represent amyloid. Postlabeling with the immunogold method of the cuprolinic blue-stained tissue proved that cuprolinic blue positive fibrils represented AA-amyloid fibrils. Therefore, it was concluded that the GAGs which appeared to colocalize with the fibrillar microanatomy of amyloid, represent a structural part of the amyloid fibrils and that the avian amyloid fibrils may be considered as a pathological proteoglycan. PMID- 16690496 TI - Lipid droplets are present in amyloid deposits in familial amyloidotic polyneuropathy and dialysis related amyloidosis. AB - It has been well documented that transthyretin (TTR) shows an affinity for lipoproteins and amyloid is deposited around adipocytes in patients with familial amyloidotic polyneuropathy (FAP). We examined the involvement of lipids in amyloid fibrils in the tissues by histopathologic methods. Sudan black B staining for frozen tissues of autopsied FAP patients and biospied dialysis related amyloidosis (DRA) patients revealed colocalization of lipids in the tissue sections where Congo red staining was positive, while no such positive staining was observed in paraffin embedded tissues. Immunohistochemical study using lipoprotein antibodies revealed that only anti-high density lipoprotein (HDL) antibody showed immunoreactivity in both types of amyloid specimens where Congo red was positively stained. Measurement of the components of lipids in the frozen cardiac samples from an FAP patient and an amyotrophic lateral sclerosis (ALS) patient revealed that concentrations of triglyceride and cholesterol in each lipoprotein, except for HDL-triglyceride, were markedly elevated in the FAP patient's material, compared to that of the ALS patient. These results suggest that interaction of amyloid fibrils with HDL may play an important role in amyloid formation in FAP. PMID- 16690497 TI - Aging and transthyretin-related amyloidosis: pathologic examinations in pulmonary amyloidosis. AB - Although aging is closely related with the onset of senile systemic amyloidosis (SSA) caused by wild-type transthyretin (TTR), the effect of aging on amyloid formation has remained unclear in familial amyloidotic polyneuropathy (FAP), caused by variant- and wild-type TTR. The aim of this study was to elucidate the effects of aging and/or other factors in FAP on amyloid formation in the lung, one of the most important target organs of amyloid deposition in SSA. Pulmonary amyloid distribution was determined using 19 autopsied lung samples from patients with FAP amyloidogenic TTR (ATTR) V30M, the most common type of FAP. Amyloid deposition was observed around the walls of the bronchi/ bronchioles, the pulmonary arteries, and the pulmonary veins, while no amyloid deposits could be found around the lymphatics. In addition, amyloid deposition in the alveolar regions was a characteristic finding in aged patients with FAP ATTR V30M (average ages of the patients with amyloid positive vs. negative: 50.55 +/- 8.75 vs. 39.75 +/- 4.17 years old, p < 0.005), similar to the finding in one SSA patient. These results suggest that aging could play an important role in the progression of pulmonary amyloid formation in FAP ATTR V30M. PMID- 16690498 TI - Natural history of thromboembolism in AL amyloidosis. AB - To determine the natural history of thromboembolism in patients with biopsy proven AL amyloidosis, medical records of 2,132 patients evaluated at Mayo Clinic between 1975 and 2000 were reviewed. Forty patients (21 male) were found to have objectively documented thromboembolism. The median age was 65 years. Four patients had a monoclonal gammopathy of undetermined significance, 20 had an unspecified plasma cell proliferative disorder, 11 had multiple myeloma, and 1 had Waldenstrom macroglobulinemia. Twelve patients had cardiac amyloid, and 20 had renal amyloid. Neither the extent of the amyloid nor the type of monoclonal protein was predictive of the development of thromboembolic events. Thromboembolism preceded the diagnosis of AL amyloidosis in 11 patients, occurred at the time of diagnosis or within 1 month after diagnosis in 11 patients, and occurred 1 month or more after diagnosis in 18 patients. Twenty-nine patients (73%) had venous thrombosis and 11 (28%) had arterial thrombosis. Eight patients (20%) died within 1 month after the thrombotic event, and 18 (45%) died within 1 year. The development of thrombosis in patients with AL amyloidosis appears to predict significant mortality within the first month and year after the event. PMID- 16690499 TI - A case of biopsy-proven leptomeningeal amyloidosis and intravenous Ig-responsive polyneuropathy associated with the Ala25Thr transthyretin gene mutation. AB - A growing body of literature has described familial leptomeningeal amyloidosis, a rare phenotype resulting from deposition of transthyretin (TTR) amyloid within the leptomeninges. We report herein the case of a patient with leptomeningeal amyloidosis presenting with hearing loss, asymmetrical polyneuropathy and sensory ataxia. This is the first Japanese case displaying TTR mutation at codon 25, replacing alanine with threonine. Neurophysiological examinations suggested demyelinating polyradiculoneuropathy, which improved dramatically after high-dose intravenous immunoglobulin treatment. Demyelinating polyneuropathy in our patient may be attributable to massive leptomeningeal amyloidosis, and no systemic organ involvement was identified. These characteristic clinical manifestations may have resulted from the Ala25Thr TTR gene mutation. PMID- 16690500 TI - Localized oral amyloidosis of the palate. AB - BACKGROUND: Amyloidosis is a rare disease with multifactorial pathogenesis. Localized amyloidosis affecting the head and neck region is an uncommon and benign process, which has almost no clinical consequences. The most reported characteristic features of localized oral amyloidosis appear as multiple soft nodules of the tongue, lip and cheek. METHODS: We report the case of a 68-year old woman suffering from a primary localized amyloidosis presenting as a purple patch on the palate. CONCLUSIONS: The presence of systemic amyloidosis or underlying plasma cell dyscrasia have to be ruled out in patients presenting with a diagnosis of amyloidosis of the oral mucosa. If a primary localized amyloidosis is proven, the surgical therapy may be useful to eliminate a functional impairment. PMID- 16690504 TI - Feto-placental communication system with the myometrium inpregnancy and parturition: the role of hormones, neurohormones, inflammatory mediators, and locally active factors. AB - Pregnancy is a unique condition in which the conceptus is allowed to implant, survive, develop, and reach a considerable organ growth and maturation within the maternal body despite the fact that it is half genetically different from the mother. Moreover, it deeply influences the overall endocrine, metabolic, and immunological functions of the recipient mother. These objectives are accomplished through the establishment of several communication systems in which a large array of substances produced by the feto-placental unit reach specific maternal target organs and/or systems and modulate their function. The myometrium is a fundamental reproductive tissue involved in pregnancy maintenance as well as in labor onset and progression and is a potential target organ for such a communication system. An appropriate regulation of myometrial function is a key condition required for pregnancy to develop physiologically until full term is reached and for labor to start. Emerging experimental and clinical evidence suggests that a very complex feto-placental biomolecular communication system exists with the myometrium and is actively operative in the control of myometrial contractility in pregnancy and parturition through the production of a continuously increasing number of substances with endocrine, paracrine, and immunoregulatory actions. PMID- 16690505 TI - Pregnancy complications and delivery outcomes of pregnant women with influenza. AB - OBJECTIVES: In a previous study, we showed that maternal influenza in pregnancy gives rise to an increase in some congenital abnormality groups. The aim of this study was to ascertain the relationship between influenza during pregnancy and pregnancy complications, and in addition, delivery outcomes particularly preterm birth and low birth weight in newborns. METHODS: The population-based large control (without any defects) data set of the Hungarian Case-Control Surveillance System of Congenital Abnormalities, in which pregnancy complications, gestational age and birth weight are medically recorded, was evaluated. RESULTS: Of 38,151 newborn infants, 1838 (4.8%) had mothers with influenza during pregnancy. The prevalence of pregnancy complications showed no difference between mothers with or without influenza during the study pregnancy. Mothers with influenza in pregnancy had a somewhat higher gestational age (0.1 week) and a lower proportion of preterm births (8.0% vs. 9.2%). These findings were reflected in the mean birth weight (+37 g) and lower proportion of low birth weight newborns (4.7% vs. 5.1%); these differences were explained by confounders. CONCLUSION: Maternal influenza during pregnancy does not increase the prevalence of pregnancy complications and unsuccessful delivery outcomes. PMID- 16690506 TI - Preeclampsia in twin gestations: association with IVF treatments, parity and maternal age. AB - OBJECTIVE: To identify maternal factors that increase the risk of preeclampsia in twin gestations and to investigate whether twins conceived by in vitro fertilization (IVF) lead to an increased risk of preeclampsia development. MATERIALS AND METHODS: A retrospective population-based cohort study of twin deliveries was performed. Maternal characteristics and perinatal outcomes were evaluated. Patients' data were obtained from a computerized database and analyzed using SPSS statistical package. RESULTS: During the study period there were 2628 twin deliveries, and of these 3.1% had severe preeclampsia and 6.16% mild preeclampsia. Patients with severe preeclampsia were more likely to be primiparous, and to have significantly higher frequency of chronic hypertension, gestational diabetes mellitus (GDM), IVF treatments, cesarean delivery, preterm delivery and twin discordancy than in the normotensive patients. Chronic hypertension, pirimiparity, twin discordancy and maternal age were independent risk factors for the development of preeclampsia. In a multivariate regression model including IVF treatment, parity and maternal age as risk factors for preeclampsia, women younger than 35 years that conceived following IVF treatments had an independent risk factor for the development of preeclampsia. CONCLUSION: IVF treatments in primiparous patients and age younger than 35 years are independent risk factors for preeclampsia. Twin discordancy is an additional independent risk factor for the occurrence of preeclampsia. PMID- 16690507 TI - The role of perinatal and intrapartum risk factors in the etiology of cerebral palsy in term deliveries in a Turkish population. AB - OBJECTIVE: To investigate the relationship between cerebral palsy (CP) and perinatal and intrapartum risk factors and markers of birth asphyxia. METHODS: This was a retrospective case-control study of 101 children with CP and 308 controls who were born at the Zeynep Kamil Hospital between 1990 and 2000. The roles of possible perinatal and intrapartum risk factors were investigated in CP development. Chi-square tests, Fisher's exact tests, Student t-tests, and regression analysis were used for statistical analysis (p < 0.05). RESULTS: The only significant perinatal risk factor was neonatal weight of <2500 grams. The CP rate in the presence of solely intrapartum risk factors was estimated as 24.7% (n = 25). In 39.6% of cases, no risk factor could be identified. When regression analysis was performed, the following factors were significant: decreased beat-to beat variability on electronic fetal monitoring (EFM) (p = 0.02), an Apgar score of <7 at 1 and 5 minutes (p = 0.02), and the necessity for neonatal intensive care unit admission (p < 0.0001). CONCLUSION: Intrapartum risk factors were significantly more frequent in the CP group (p < 0.01). An increased cesarean section rate could not prevent CP, suggesting that the hypoxic insult that causes CP might be of chronic onset. The development of diagnostic tests to detect non reassuring fetal status in its intrauterine life and interventions at appropriate times may decrease the CP rate. PMID- 16690508 TI - Ponderal index for discrimination between symmetric and asymmetric growth restriction: percentiles for neonates from 30 weeks to 43 weeks of gestation. AB - AIMS: The ponderal index describes body proportionality at birth thus distinguishing symmetric from asymmetric growth restriction. We aimed to develop ponderal index percentiles for preterm and term neonates born in a European population. METHODS: Auxologic data were obtained from neonates born from January 1990 to December 1998 from the datasets reported to the perinatal quality assurance system of the Federal State of Hesse, Germany. We excluded data from neonates with lethal malformations, with chromosomal aberrations, from multiple births, from neonates with uncertain gestational age, and from neonates of a gestational age of less than 30 completed weeks. We calculated the weekly 5th, 10th, 25th, 50th, 75th, 90th, and 95th ponderal index percentile values. RESULTS: A total of 480,841 neonates (233,662 females and 247,179 males) were included. Charts and tables of ponderal index values show percentiles for males, females, and for the total group. There were no significant differences between boys and girls. CONCLUSION: Our data offer the ability to refer a neonate's body proportionality to updated percentiles. The percentiles allow the discrimination between symmetric and asymmetric growth restriction in preterm and term infants. PMID- 16690509 TI - The need for a new outlook on labor monitoring. AB - Fetal heart rate monitors, including the newer pulse-oximetry and STAN monitors, are designed to detect fetal distress that affects less than 1% of women in labor. Non-progressive labor is a much more common disorder than fetal distress, with approximately 50% of women in labor requiring oxytocin. Current technology assessing labor progress is subjective and inaccurate. There is a need for objective and accurate technology to measure labor progress and the effect it may have on managing labor and, specifically, non-progressive labor. PMID- 16690510 TI - Computerized analysis of fetal heart rate variability using the matching pursuit technique as an indicator of fetal hypoxia during labor. AB - OBJECTIVE: To determine whether the computerized analysis of fetal heart rate variability with the new matching pursuit technique can indicate fetal distress during labor. STUDY DESIGN: Eighty women were studied during the intrapartum period with external cardiotocography. In all cases, cord arterial pH and 5-min Apgar Scores were evaluated. Six cases that presented large segments of missing data were excluded from the study. The remaining 74 women were divided into two groups; 32 women with normal (Group A) and 42 women with non-reassuring FHR tracings (group B). Group B was divided in subgroup BI, including 24 women with pH > 7.20, and BII, including 18 women with pH < 7.20. In order to evaluate the FHR fluctuations, in different frequency ranges, we applied an adaptive time frequency method, called Matching Pursuit. We estimated the power of the FHR signal in four frequency ranges. RESULTS: The 5-min Apgar Scores were significantly lower in both subgroup BI and subgroup BII (p = 0.003 and p = 0.003 respectively). The Low Low Frequency (LLF) parameter appears to recognize better the cases with lower pH (sensitivity 78.5%, specificity 52.3%) than the cases with non-reassuring FHR (66.6%, 56.2). The sensitivity and specificity of the Very Low Frequency (VLF) parameter were 72.2% and 59% respectively in recognizing the cases with lower pH and 64.2% and 53.1% in recognizing non-reassuring FHR. CONCLUSION: Fetal hypoxia during labor can be recognized using the MP technique for the analysis of FHR signal power in the VLF and LLF frequency ranges. Since the analysis is feasible in real-time, it can be a useful tool for the intrapartum evaluation of fetal well-being. PMID- 16690511 TI - Magnesium sulfate effectively reduces blood pressure in an animal model of preeclampsia. AB - OBJECTIVE: We tested the ability of magnesium sulfate to reduce hypertension and neonatal growth retardation in an animal model of preeclampsia. STUDY DESIGN: On day 17 of pregnancy, osmotic minipumps were inserted subcutaneously to continuously deliver either vehicle (saline control group), or N-nitro-L-arginine methyl ester (L-NAME) (50 mg/kg/day), or L-NAME (50 mg/kg/day) in combination with magnesium sulfate (60 mg/kg/day). Prior to insertion, blood pressure and heart rate were monitored with a pneumatic tail cuff device. Blood pressure measurements were repeated on days 18, 20, and 21 of pregnancy. Blood was obtained on days 17 and 21, along with urine, to assess magnesium levels and degree of proteinuria. Pups were weighed and measured at 48 hours postpartum. RESULTS: Rats receiving L-NAME developed hypertension within 24 hours of implantation (108 +/- 3.9 vs. 123 +/- 3.4 mmHg, p < 0.05). Magnesium sulfate, given along with L-NAME did not prevent mean blood pressure from increasing, but reduced it by day 21 compared to L-NAME given alone (107 +/- 3.4 vs. 122 +/- 8.7 mmHg, respectively, p < 0.05). Magnesium sulfate reduced neonatal growth retardation by improving the weight of the pups compared to pups from maternal rats given L-NAME alone (6.1 +/- 0.1 vs. 5.2 +/- 0.3 grams, respectively, p < 0.05). CONCLUSION: Maternal magnesium sulfate reduces blood pressure and increases neonatal size compared to L-NAME without magnesium. These findings support a beneficial effect of magnesium in preeclampsia. PMID- 16690512 TI - A systematic review of intentional delivery in women with preterm prelabor rupture of membranes. AB - OBJECTIVE: To evaluate the effect of intentional delivery versus expectant management in women with preterm prelabor rupture of membranes (PPROM). METHODS: We searched electronic databases and trials registries, contacted experts, and checked reference lists of relevant studies. Studies were included if they were randomized controlled trials comparing intentional delivery versus expectant management after PPROM, the gestational age of participants was between 30 and 36 weeks, and the study reported one of several pre-determined outcomes. RESULTS: Four studies were included in the meta-analysis. No difference was found between intentional delivery and expectant management in neonatal intensive care unit (NICU) length of stay (LOS) (weighted mean difference (WMD) -0.81 day, 95% confidence interval (CI) -1.66, 0.04), respiratory distress syndrome (risk difference (RD) -0.01, 95% CI -0.07, 0.06), and confirmed neonatal sepsis (RD 0.01, 95% CI -0.05, 0.04). One study found a significantly lower incidence of suspected neonatal sepsis among the intentional delivery group (RD -0.31, 95% CI 0.50, -0.12; number needed to treat (NNT) 3, 95% CI 2, 8). Maternal LOS was significantly shorter for the intentional delivery group (WMD -1.39 day, 95% CI 2.03, -0.75). There was a significant difference in the incidence of clinical chorioamnionitis favoring intentional delivery (RD -0.16, 95% CI -0.23, -0.10; NNT 6, 95% CI 5, 11). There was no significant difference in the incidence of other maternal outcomes, including cesarean section (RD 0.05, 95% CI -0.01, 0.11). CONCLUSIONS: Intentional delivery may be favorable to expectant management for some maternal outcomes (chorioamnionitis and LOS). There is insufficient evidence to suggest that either strategy is beneficial or harmful for the baby. Large multicenter trials with primary neonatal outcomes are required to assess whether intentional delivery is associated with less neonatal morbidity. PMID- 16690513 TI - Fetal outcome following roxithromycin exposure in early pregnancy. AB - OBJECTIVE: Because very little information exists on the fetal safety of roxithromycin, we aimed to extend the knowledge on fetal outcome in pregnant women who were exposed to roxithromycin in early pregnancy. METHODS: Twenty pregnant women inadvertently exposed to roxithromycin during early pregnancy were identified and prospectively followed-up. For comparison, 170 pregnant women matched by age and gravidity, not being exposed to any potential teratogenic agent during pregnancy, were recruited as controls. All gestations were confirmed by ultrasound examination, and participants were followed-up until delivery. Newborns were examined by a neonatologist. RESULTS: Of 20 pregnant women exposed to roxithromycin during early pregnancy, information was obtained from 17 cases. The median dose of roxithromycin to which pregnant women were exposed was 300 mg/day (range 300-450 mg/day) and exposure occurred at a mean of 4.0 (range 2.8 17.6) weeks. Mean gestational age at delivery was 39.2 weeks in the exposed group and 39.4 in the controls (p = 0.6). Birth weight of babies exposed in utero to roxithromycin was not different to controls. We did not observe any major malformation in the exposed group whereas three (1.8%) occurred in the control group. CONCLUSIONS: . Despite the limitations of the study due to the small sample size, roxithromycin appears not to be a major teratogen. PMID- 16690514 TI - Influence of parity on second-trimester uterine artery Doppler waveforms in twin pregnancy. AB - We performed uterine artery Doppler velocimetry in 16 nulliparous and 16 parous women with non-preeclamptic singleton gestations and 16 nulliparous and 16 parous women with dichorionic twin gestations at 17-18 and 26-27 weeks of gestation. In both singleton and twin pregnancies, the average pulsatility index (PI) in nulliparous women was significantly higher than that in parous women at 17-18 weeks of gestation. At 26-27 weeks of gestation, however, there were no significant differences in PI between nulliparous and parous women with either singleton or twin pregnancies. During the early second-trimester of non preeclamptic singleton and twin pregnancies, parity has a significant effect on uterine artery blood flow. PMID- 16690515 TI - Unrelated cord blood allogeneic stem cell transplantation for MDS. PMID- 16690516 TI - Environmental factors and childhood acute leukemias and lymphomas. AB - This review considers recent studies regarding the role of environmental factors in the etiology of childhood leukemia and lymphoma. Potential environmental risk factors identified for childhood leukemia include exposure to magnetic fields of more than 0.4 micro Tessla, exposure to pesticides, solvents, benzene and other hydrocarbons, maternal alcohol consumption (but only for certain genotypes), contaminated drinking water, infections, and high birth weight. The finding of space-time clustering and seasonal variation also supports a role for infections. There is little evidence linking childhood leukemia with lifetime exposure to ionizing radiation although fetal exposures to X-rays are associated with increased risk. Breast-feeding, consumption of fresh fruit and vegetables and having allergies all appear to be protective. Burkitt lymphoma (BL) is confined to areas of the world where malaria is endemic, with the additional involvement of the Epstein-Barr virus (EBV) as a co-factor. Environmental risk factors suggested for other types of non-Hodgkin lymphoma (NHL) include exposure to ionizing radiation (both lifetime and antenatal), pesticides, and, in utero exposure to cigarette smoke, benzene and nitrogen dioxide (via the mother). Hodgkin lymphoma (HL) is especially associated with higher levels of socioeconomic deprivation, but breast-feeding seems to confer lower risk. This is consistent with an infection or immune-response mediated etiology for HL. PMID- 16690517 TI - The efficacy of unrelated cord blood transplantation for adult myelodysplastic syndrome. AB - Although allogeneic stem cell transplantation from a human leukocyte antigen (HLA)-identical related donor offers a potential cure for patients with myelodysplastic syndrome (MDS), a suitably matched related donor is unavailable for approximately two thirds of patients. Recently, umbilical cord blood from unrelated donors have been used as an alternative stem cell source for adult patients with MDS. Here, we updated the results of unrelated cord blood transplantation (CBT) after myeloablative conditioning for 22 adult patients with MDS. Diagnosis at transplantation included refractory anemia (RA) (n = 3), refractory anemia with excess blasts (RAEB) (n = 2), RAEB-t (n = 2), and MDS related secondary acute myeloid leukemia (AML) (n = 15). All patients were treated with total body irradiation (12 Gy), cytosine arabinoside (Ara-C) and cyclophosphamide followed by unrelated HLA-mismatched CBT. The median age was 40 years (range, 19 - 51 years), the median weight was 54.5 kg (range, 43 - 75 kg), and the median number of cryopreserved nucleated cells was 2.43 x 10(7)/kg (range, 1.82 - 4.10 x 10(7)/kg). Twenty one patients had myeloid reconstitution and the median time to more than 0.5 x 10(9)/l absolute neutrophil count was 22.5 days. A self-sustained platelet count more than 50 x 10(9)/l was achieved in 19 patients at a median time of 49 days. Acute GVHD above grade II occurred in seven of 21 evaluable patients and chronic GVHD in 16 of 19 evaluable patients. Among 16 chronic GVHD patients, in eight patients the disease was extensive. Seventeen patients are alive and free of disease at between 371 and 2562 days after transplantation. With a median follow-up of 1505 days, the probability of disease free survival at 4 years was 76.0%. These results suggest that adult MDS patients without suitable related or unrelated bone marrow donors should be considered as candidates for CBT. PMID- 16690518 TI - Aggressive non-Hodgkin's lymphoma: concomitant evaluation of interleukin-2, soluble interleukin-2 receptor, interleukin-4, interleukin-6, interleukin-10 and correlation with outcome. AB - The purpose of this study was to assess the prognostic value of a large panel of cytokines in aggressive non-Hodgkin's lymphoma (NHL) and to confront it to parameters of the International Prognostic Index (IPI). It investigated the concomitant determination of interleukin-2 (IL-2), soluble interleukin-2 receptor (sIL-2R), interleukin-4 (IL-4), interleukin-6 (IL-6) and interleukin-10 (IL-10) on a uniform population of 116 previously untreated patients. Commercially available enzyme-linked immunoassay kits were used for cytokines measurements. Results were correlated with complete remission (CR), overall survival (OS) and failure free survival (FFS). In univariate analysis, sIL-2R and IL-6 demonstrated prognostic significance for CR (p = 0.016 and p = 0.048), OS (p = 0.0011 and p = 0.0387) and FFS (p = 0.0001 and p = 0.0363), but multi-variate analysis failed to demonstrate an independent prognostic significance. In the intermediate group risk defined by IPI, patients presenting high level of sIL-2R or IL-6 demonstrated lower CR rate and survival than those with low level. In conclusion, sIL-2R and IL-6 serum levels are elevated in high grade NHL and are correlated to CR, OS and FFS, but this study did not support their independent prognostic value. However, sIL-2R and IL-6 measurements may improve risk assignment by IPI and allow a better prognostic evaluation of patients with intermediate prognosis NHL. PMID- 16690519 TI - Dendritic cells are significantly reduced in non-Hodgkin's lymphoma and express less CCR7 and CD62L. AB - Despite the lack of tumor control, infiltration of immune cells has been demonstrated for several malignancies including non-Hodgkin's lymphoma. Since dendritic cells play a pivotal role in the initiation and control of the immune response, the frequency and phenotype of recently described sub-types of dendritic cells in non-Hodgkin's lymphoma were characterized. Myeloid and plasmacytoid dendritic cells were analysed in 55 non-Hodgkin's lymphoma and 33 reactive lymph nodes by flow cytometry and immunohistochemistry. Overall frequency of dendritic cells in reactive lymph nodes was higher than in non Hodgkin's lymphoma while the pDC/mDCs ratio was comparable. The low frequency of dendritic cells in infiltrated lymph nodes was confirmed by immunohistochemistry; however, no significant difference in the distribution within lymphoid and tumor tissue was detected. For further characterization of the dendritic cells in non Hodgkin's lymphoma, the expressions of adhesion molecules, costimulatory molecules, chemokine receptors and activation markers were assessed. Interestingly, a significantly decreased expression of CD62L and CCR7, receptors necessary for homing to lymph nodes, was identified in dendritic cells in non Hodgkin's lymphoma, potentially explaining the lack of these cells. Taken together, dendritic cells are phenotypically altered and reduced in number in NHL, potentially contributing to the loss of tumor control in these patients. PMID- 16690520 TI - Fertility status among women treated for aggressive non-Hodgkin's lymphoma. AB - In young women treated for intermediate-high-grade non-Hodgkin's lymphoma with CHOP (cyclophosphamide, adriamycin, oncovine and prednisone), there is insufficient data concerning gonadotoxicity or the need for fertility-preserving measures. The aim of the present study was to evaluate the fertility status in the first complete remission of women who were treated for aggressive non Hodgkin's lymphoma. A cohort of 36 women with aggressive non-Hodgkin's lymphoma in first remission, who were treated in five university-affiliated hospitals in Israel, was evaluated. All women were aged younger than 40 years at diagnosis and received frontline protocols, including cyclophosphamide and adriamycin, mostly CHOP. Menstrual cycle characteristics, as well as pregnancies before the diagnosis, during treatment and in first complete remission, were evaluated. The patients' mean age at the diagnosis was 28 +/- 7 years (range 17 - 40 years). All patients were treated with chemotherapy, although 10 patients received additional radiotherapy. Follow-up time at first complete remission was 84 +/- 48 months. Before diagnosis, all patients had menstrual cycles, which were regular in 31 (86%). Three patients received gonadtropin-releasing hormone analogs, whereas nine received contraceptive pills together with cytotoxic treatment. During treatment, 18 patients (50%) had amenorrhea, six (17%) had irregular menstrual cycles, and 12 (33%) continued their regular cycles. All but two women resumed menses in the first complete remission, and these were regular in 22 (61%) patients. In 63% of patients, the menstrual cycle recovered within 3 months of the discontinuation of chemotherapy. Eighteen patients (50%) became pregnant during the first complete remission. There was no significant difference between those patients who received fertility-preserving measures versus the remainder concerning regular menstrual cycles recovery or pregnancies. The two patients who developed amenorrhea were 40 years old at the time of diagnosis. In conclusion, the rate of gonadal dysfunction is very low among young, CHOP treated, non Hodgkin's lymphoma female patients. Fertility-preserving techniques are not needed for women aged younger than 40 years and should probably be reserved for those who are at high risk for gonadal toxicity. PMID- 16690521 TI - Treatment with yttrium 90 ibritumomab tiuxetan at early relapse is safe and effective in patients with previously treated B-cell non-Hodgkin's lymphoma. AB - Yttrium 90 ((90)Y) ibritumomab tiuxetan (Zevalin), a radiolabeled monoclonal antibody against the CD20 antigen, is indicated for the treatment of patients with relapsed or refractory low-grade, follicular, or transformed B-cell non Hodgkin's lymphoma (NHL), including patients with rituximab-refractory follicular NHL. Data on 211 patients treated in four clinical trials were analysed to compare the efficacy and safety of (90)Y ibritumomab tiuxetan when it was used after the first relapse of NHL and when it was used after two or more prior therapies. Sixty-three patients (30%) were treated with (90)Y ibritumomab tiuxetan after their first relapse and 148 (70%) after two or more prior therapies. Demographics, disease characteristics and the frequency of adverse events were similar in all groups, with the exception of a higher rate of marrow involvement in first-relapse patients than in patients with two or more prior therapies (57% vs. 39%; P < 0.05). The complete response rate [confirmed (CR) and unconfirmed (Cru)] was higher in first-relapse patients (49% vs. 28%; P < 0.01), and the median time to progression (TTP) was longer (12.6 vs. 7.9 months; P < 0.05). In patients with follicular NHL, the differences were even more pronounced (CR/CRu: 51% vs. 28%; P < 0.01; TTP: 15.4 vs. 9.2 months; P < 0.05). (90)Y ibritumomab tiuxetan has substantial clinical benefits as a second-line therapy, especially in patients with follicular NHL. The quality of disease remissions obtained when (90)Y ibritumomab tiuxetan is administered after first relapse appears to be comparable with that observed with most chemotherapy regimens in first-relapse patients. PMID- 16690522 TI - Procarbazine for non-Hodgkin's lymphoma. AB - Procarbazine hydrochloride is an oral alkylating agent with activity against lymphoma. It is most commonly used in the treatment of Hodgkin's disease. The use of procarbazine-containing chemotherapeutic regimens in non-Hodgkin's lymphoma fell out of favor with the advent of CHOP. We report two patients with relapsed and/or refractory follicular lymphoma that achieved a complete and durable remission with a prolonged course of daily procarbazine. PMID- 16690523 TI - Mitoxantrone, vinblastine and CCNU: long-term follow-up of patients treated for advanced and poor-prognosis Hodgkin's disease. AB - Advanced-stage or relapsed/refractory Hodgkin's disease (HD) has a poor prognosis despite aggressive chemotherapy regimens and the use of high-dose therapy with autologous stem cell support. Mitoxantrone, vinblastine and CCNU (lomustine) (MVC) combines the most effective chemotherapeutic agents of previous regimens for poor prognosis HD, and eliminates marginally active agents with unnecessary toxicities, such as bleomycin and dacarbazine. Sixty-eight patients with HD (23 newly diagnosed and 45 with relapsed/refractory disease, one patient treated both de novo and years later in relapse) were treated with the MVC regimen (mitoxantrone 8 mg/m(2)/day i.v. days 1 - 3; vinblastine 8 m/m(2)/day days 1 and 22; and CCNU (lomustine) 100 mg/m(2) on day 1, repeated at 6 - 8 weeks) in a single-arm Phase II study. All patients responded to treatment in the newly diagnosed group (overall response = 100%). The median response duration was not reached, but was in the range 7.6 - 180 + months, and median survival was 94 months. Eleven complete responses are ongoing at 39 - 180 + months. In the previously-treated patients, 41 responded to MVC (OR = 91%). The median response duration for this group was 11 months, and the median survival was 34 months after initiating MVC. Four secondary myeloid leukemias occurred, three in de novo, and one in the relapsed/refractory group, at a median follow-up of 14 years. MVC regimen for HD is highly active, for both de novo and relapsed/refractory disease, with high response rates and survival that compare favourably with the results obtained by high-dose therapy with stem-cell transplantation. Although significant, the toxicities associated with this regimen were manageable. PMID- 16690524 TI - Neutropenia and febrile neutropenia in patients with Hodgkin's lymphoma treated with doxorubicin (Adriamycin), bleomycin, vinblastine and dacarbazine (ABVD) chemotherapy. AB - When uncomplicated neutropenia during doxorubicin (Adriamycin), bleomycin, vinblastine and dacarbazine (ABVD) chemotherapy for the treatment of Hodgkin's lymphoma is encountered, it is unclear whether or not treatment should be modified. In the present study, we determined the incidence of neutropenia, febrile neutropenia, and the relationship of febrile neutropenia to grade III/IV neutropenia and dose modification, in a large university patient population. We reviewed the charts of patients diagnosed with Hodgkin's lymphoma between 1 January 1990 and 31 December 2002 who were treated with ABVD chemotherapy, and seen at the University of Iowa with complete diagnosis, staging, and treatment dosing records. Adequate data was available on 894 treatments in 81 patients with Hodgkin's lymphoma treated with ABVD chemotherapy. Grade III/IV neutropenia was present on the scheduled day of treatment in 187 (20.9%) treatments in 64 (79%) patients. Grade III/IV neutropenia was most common at cycle 1 day 15. Febrile neutropenia developed nine times in eight patients, and eight episodes of febrile neutropenia developed when the treatment-day absolute neutrophil count (ANC) > or =1000. Dose delay of >4 days and/or dose reduction to <80% of original doxorubicin dose following grade III/IV neutropenia occurred in 29 of 187 treatments, with no episodes of febrile neutropenia. With grade III/IV neutropenia on the day of therapy, 158 treatments were administered without dose reduction or dose delay with one subsequent episode of febrile neutropenia. Neutropenia during ABVD is common, and dose modification for uncomplicated neutropenia on the day of treatment may not reduce the risk of febrile neutropenia. It may be possible to maintain dose intensity in the face of uncomplicated neutropenia during ABVD therapy. PMID- 16690525 TI - Expression of ice, bcl-2, c-myc and p53 in different bone marrow cell populations from patients with diffuse large B-cell lymphoma. AB - We have previously reported functional alterations in vitro in the hematopoietic compartment of patients with diffuse large B-cell lymphoma (DLBCL). In the present study, we assessed the presence of molecular alterations in hematopoietic cells derived from DLBCL marrow. Accordingly, the expression of four genes (i.e. ice, bcl-2, c-myc and p53) was assessed both, at the mRNA and protein levels, in three cell populations: (i) population I, consisting of morphologically recognizable precursor and mature cells; (ii) population II, enriched for CD34+ Lineage-negative (Lin-) cells; and (iii) population III, enriched for CD34+ CD38- Lin- cells. By using a multiplex reverse transcriptase-polymerase chain reaction system, we observed reduced expression of bcl-2 in population I, and c-myc in populations I and II from lymphoma marrow compared to their normal counterparts. On the other hand, expression of ice and p53 was not significantly different when comparing normal and DLBCL samples. At the protein level, all four molecules were expressed in a higher proportion of samples from DLBCL patients than in marrow samples from normal subjects. Interestingly, these proteins were expressed predominantly in primitive cells (population III), whereas the proportion of positive samples was reduced in population II, and even more in population I. Taken together, our results indicate that, in DLBCL, molecular alterations are present in hematopoietic cells from bone marrow, including morphologically recognizable precursor and mature cells, as well as primitive hematopoietic progenitors (CD34+ cells). To date, the physiological implications of these alterations are still unclear, and further studies should be undertaken to address this issue. PMID- 16690526 TI - Autologous dendritic cells pulsed with eluted peptide as immunotherapy for advanced B-cell malignancies. AB - We have studied the feasibility, safety and efficacy of vaccination with autologous dendritic cells pulsed with eluted peptide in patients with advanced low-grade B-cell malignancies. This study demonstrates that autologous dendritic cell vaccines can be successfully produced from patients with advanced disease and be delivered without significant toxicity. Furthermore, we have demonstrated immunological and clinical responses in two of ten patients treated. These results provide further evidence for the use of immunotherapy in the management of B-cell malignancies, but also suggest that sustained responses may only be possible in patients with low bulk disease early in the disease course. PMID- 16690527 TI - Recommendation of the use of myeloblast percentage among non-erythroid cells instead of percentage among total nucleated cells for therapeutic response assessment in acute erythroid leukemia. AB - The diagnostic criteria of acute erythroid leukemias (AEL) has been revised by WHO in 2001. The National Cancer Institute (NCI) published a set of standardized diagnostic and response criteria for acute myeloid leukemia in 1990, which was revised in 2003. The aim of the present study was to establish the best criteria for therapeutic response assessment in the newly classified AEL and evaluate patient outcomes. Fifty-two patients with AEL as defined by the new WHO criteria were evaluated in this study. The following seven indices for therapeutic response assessment were evaluated: (i) NCI criteria (myeloblast percentage among total nucleated cells (TNC) and cellularity); (ii) myeloblast percentage among non-erythroid cells (NEC) and cellularity; (iii) erythroid series percentage among TNC; (iv) pronormoblast percentage among erythroid cells; (v) ratio of pronormoblasts and blasts; (vi) maturation arrest index; and (vii) disappearance of erythroid dysplasia. Complete remission (CR) patients with <5% of myeloblast/NEC (NEC-CR) showed significantly longer overall survival periods (mean 55.8 months) compared to CR patients with >5% myeloblast/NEC (mean 11.7 months, P = 0.006). NEC-CR patients also had longer event-free survival (median 16.4 months) compared to patients with >5% and <20% of myeloblast/NEC (median 6.2 months) (P = 0.044). The other indices for therapeutic response assessment are not significant for predictability of relapse and outcomes. Therefore, we recommend that the myeloblast percentage among NEC be used instead of myeloblast percentage among TNC for therapeutic response assessment in AEL. PMID- 16690528 TI - The role of post-remission chemotherapy for older patients with acute myelogenous leukemia. AB - Standard practice in older patients with acute myeloid leukemia (AML) is induction chemotherapy (ICT) followed by post-remission chemotherapy (PRT). We previously reported a median disease-free survival (DFS) and overall survival (OS) for patients in complete remission (CR) of 7.5 and 13.5 months, respectively, in 30 older patients treated with standard ICT and PRT (study A). We designed a subsequent trial excluding PRT (study B). Forty patients with AML age > or =60 years were treated with ICT consisting of standard dose cytosine arabinoside and mitoxantrone followed with granulocyte-macrophage colony stimulating factor subcutaneously starting day 11 if the bone marrow aspirate and biopsy was hypocellular. Median age was 68 years. Myelodysplasia preceded AML in 37% of patients. Favourable, normal, and unfavourable karyotypes were seen in 7.5%, 55%, and 37.5% of patients, respectively. Twenty-one patients (52.5%) achieved CR. Median DFS and OS for patients in CR were 6.2 and 10.8 months, respectively. Study A and B differed by the addition of PRT in study A. However, DFS and OS did not differ significantly between patients treated in study A or study B (P = 0.21 and P = 0.15, respectively). PRT has not clearly improved survival in older patients with AML, and therefore the routine addition of chemotherapy to older patients in complete remission is not indicated. PMID- 16690529 TI - Sequential response-adapted induction and consolidation regimens idarubicin/cytarabine and mitoxantrone/etoposide in adult acute myelogenous leukemia: 10 year follow-up of a study by the Canadian Leukemia Studies Group. AB - PURPOSE: The Canadian Leukemia Studies Group (CLSG) sought to test the safety and efficacy of response-adapted, non-cross resistant chemotherapy in de novo acute myeloid leukemia (AML). The combinations of idarubicin 12 mg/m(2)/d on days 1 - 3 and Ara-C (200 mg/m(2)/d) on days 1 - 7 (IDAC) followed by mitoxantrone 10 mg/m(2)/day, and etoposide 100 mg/m(2)/day, on days 1 - 5 (NOVE) were used according to patient response to induction and consolidation. PATIENTS AND METHODS: In this multi-centre open-label phase II study, 140 patients up to age 80 were given induction with IDAC. Patients were entered between March 1993 and August 1995. If patients had persistent blasts at day 14 or on recovery, they were given NOVE. As consolidation, patients achieving complete remission (CR) with IDAC were given 1 further cycle of IDAC and 1 cycle of NOVE. Patients achieving CR after NOVE were given 2 further cycles of NOVE. RESULTS: 76% of all patients achieved remission after IDAC +/- NOVE, 81% in patients aged < or =60 years and 67% in patients aged >60. Overall, induction mortality was 11% and toxicity was similar to other cooperative group studies. Median follow-up was 104.0 months with 95% CI: (100.0, 105.2). Median overall survival (OS) in responding patients < or =60 was not reached: of the 79 responders < or =60, 35 died. The median disease free survival (DFS) in these responding patients was 22.7 (14.9, na) months. Median OS and DFS in responding patients >60 was 10.0 (7.3, 15.2) months and 7.5 (6.2, 15.2) months, respectively. CONCLUSION: The results of this trial are very encouraging and suggest that there may be long term benefit to this method. On the basis of these results, a randomized phase III trial has been performed. PMID- 16690530 TI - Epstein-Barr virus infection is not the sole cause of high prevalence for Hodgkin's lymphoma in Saudi Arabia. AB - The age-adjusted incidence of Hodgkin's lymphoma (HL) is markedly higher in Saudi Arabia than in the USA, and accounts for 10.5% of all neoplasias in children aged 15 years or older in Saudi Arabia. Epstein-Barr virus (EBV) infection has been suspected to cause high HL incidence in developing countries. To investigate the role of EBV for the high frequency of HL in Saudi Arabia, we analysed 169 HLs from Saudi Arabia and 30 HLs from Europe for EBV infection by in situ hybridization with fluorescence in-conjugated EBV on tissue microarray sections. All Saudi Arabian and European HLs were analysed in one experiment under identical conditions. Unexpectedly, our data show only minor, insignificant differences in EBV infection rates between Saudi Arabian (42 out of 147 informative cases 28.6%) and European HL (nine out of 30 informative cases; 30%; P = 0.8752). Within the Saudi Arabian population, EBV infection was most frequently seen in mixed cellularity HL (52.4%). This was significantly more frequent than in nodular sclerosing HL (26.1%; P = 0.0236). EBV positivity was unrelated to patient prognosis. In conclusion, our data strongly suggest that EBV is not the main cause for the high prevalence of HL in Saudi Arabia. This would be consistent with a major role of genetic susceptibility genes for HL in these populations. The Saudi Arabian population, with high consanguinity and large families, would prove ideal for identifying HL susceptibility genes. PMID- 16690531 TI - Acute leukemia of dendritic cell lineage in childhood: incidence, biological characteristics and outcome. AB - CD4+ CD56+ malignancies have only recently been related to dendritic cell (DC) lineage. The few cases described, mostly adults and elderly, typically present with cutaneous lesions, followed by disseminated tumor localizations within a few months, with a generally very aggressive course and fatal outcome, despite the different therapeutic approaches employing chemotherapy and/or radiotherapy. Considering that leukemias in childhood and in adults are different diseases, we describe three pediatric cases to help compare the biological characteristics, immunophenotype, clinical features, treatment response and incidence of this disease in both age groups. From a total 1363 new patients with acute leukemia (AL), we report three cases with blasts of French - American - British L2 morphology, an absence of the most specific markers for myeloid, T or B lineage and lacking CD34, which led us to evaluate the blasts with an extensive panel of antibodies, including those related to the other putative pathways of lymphoid differentation: natural killer and DC. The cells expressed CD4, CD56, HLA-DR, BDCA-2 and BDCA-4. None of our cases presented with skin involvement. All three children showed good response to acute lymphoblastic leukemia (ALL) protocols, achieving complete remission even when one of the patients relapsed and received an allogeneic transplant. These findings, in spite of the small number of patients, suggest that the clinical course in children might be less aggressive, and that regular ALL protocols would be effective. We emphasize the importance of including antibodies for DC lineage in cases of CD34(-) unclassifiable AL to further characterize these rare cases (0.22%), considering that the tumor cell affiliation to DC lineage relies exclusively on immunophenotypic criteria. PMID- 16690532 TI - Vitamin D receptor (VDR) localization in human promyelocytic leukemia cells. AB - Although vitamin D analogs are known to induce the differentiation of the HL-60 promyelocytic leukemia cells, the effect of vitamin D analogs on the distribution of vitamin D receptor (VDR) in these cells is not well studied. This report showed, by confocal microscopy, that VDR mainly resided in the cytoplasm in the absence of VDR ligands. When cells were treated with 19-nor-1alpha,25-(OH)(2)D(2) or 1,25(OH)(2)D(3), VDR moved from the cytoplasm into the nucleus in a time dependent manner. VDR could be observed in the nucleus as early as 6 h after drug treatment and was still observed in the nucleus 3 days after one single addition of 100 nM 19-nor-1alpha,25-(OH)(2)D(2) or 1,25(OH)(2)D(3). The VDR protein level was significantly increased by 19-nor-1alpha,25-(OH)(2)D(2) or 1,25(OH)(2)D(3) in a dose-dependent manner, while the VDR mRNA level was not affected by either compound. These results suggest that binding of vitamin D analogs to VDR induced receptor translocation into the nucleus, which stabilizes the receptor, resulting in an accumulation of the VDR protein. PMID- 16690533 TI - Cotylenin A-induced differentiation is independent of the transforming growth factor-beta signaling system in human myeloid leukemia HL-60 cells. AB - Cotylenin A, which has been isolated as a plant growth regulator, potently induces the differentiation of human myeloid leukemia cells. Treatment of HL-60 cells with a combination of transforming growth factor (TGF)-beta and 1alpha, 25 dihydroxyvitamin D(3) (VD3) resulted in increased differentiation compared to separate treatments, but TGF-beta did not affect the cotylenin A-induced differentiation of HL-60 cells. It is possible that the signal transduction pathway used by cotylenin A for inducing the differentiation of leukemia cells is the same as that used by TGF-beta. However, cotylenin A did not affect the expression of TGF superfamily or Smad genes in HL-60 cells. Treatment with neutralizing anti-TGF-beta antibody or an inhibitor of TGF-beta signaling did not inhibit cotylenin A-induced differentiation, although VD3-induced differentiation was significantly suppressed by these treatments. The subcellular distribution of Smad3 was also unaffected by cotylenin A. These results suggest that the cotylenin A-induced differentiation of leukemia cells is independent of the TGF beta signaling system, although TGF-beta acts as an autocrine mediator of the growth arrest and differentiation of leukemia cells induced by VD3 and other inducers. PMID- 16690534 TI - Platelet aggregation profile as a marker of hydroxyurea bioavailability through nitric oxide generation in chronic myelogenous leukemia. AB - Platelet aggregation profiles were studied in chronic myelogenous leukemia patients who were undergoing hydroxyurea therapy. Nitric oxide (NO) generation induced by hydroxyurea was measured from the altered aggregatory response, in which the platelet suspension exhibits a de-aggregatory behaviour. NO caused platelet de-aggregation by generation of cyclic guanidine monophosphate through the activation of soluble guanylate cyclase (SGC). The fact that the observed response is specific to NO was confirmed by the reversal of the de-aggregatory behaviour in the presence of (1)H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), an inhibitor of SGC. Among the subjects studied, one subset showed an hydroxyurea induced de-aggregatory effect that was inhibited by ODQ, whereas another subset did not show any such effect. The observed inter-individual variability in platelet aggregometric response after the ingestion of drugs may be an indicator for NO generation from hydroxyurea, and this may help to explain the drug efficacy encountered in such cases. PMID- 16690535 TI - Unusual reaction to Rituximab with intra-vascular hemolysis, rhabdomyolysis, renal failure and bone marrow necrosis. PMID- 16690537 TI - Gene-gene, gene-environment & multiple interactions in colorectal cancer. AB - This review comprehensively evaluates the influence of gene-gene, gene environment and multiple interactions on the risk of colorectal cancer (CRC). Methods of studying these interactions and their limitations have been discussed herein. There is a need to develop biomarkers of exposure and of risk that are sensitive, specific, present in the pathway of the disease, and that have been clinically tested for routine use. The influence of inherited variation (polymorphism) in several genes has been discussed in this review; however, due to study limitations and confounders, it is difficult to conclude which ones are associated with the highest risk (either individually or in combination with environmental factors) to CRC. The majority of the sporadic cancer is believed to be due to modification of mutation risk by other genetic and/or environmental factors. Micronutrient deficiency may explain the association between low consumption of fruit/vegetables and CRC in human studies. Mitochondrial modulation by dietary factors influences the balance between cell renewal and death critical in colon mucosal homeostasis. Both genetic and epigenetic interactions are intricately dependent on each other, and collectively influence the process of colorectal tumorigenesis. The genetic and environmental interactions present a good prospect and a challenge for prevention strategies for CRC because they support the view that this highly prevalent cancer is preventable. PMID- 16690538 TI - An evaluation of the biological and toxicological properties of Aloe barbadensis (miller), Aloe vera. AB - Aloe barbadensis (Miller), Aloe vera, has a long history of use as a topical and oral therapeutic. The plant is the source of two products, gel and latex, which are obtained from its fleshy leaves. Aloe vera products contain multiple constituents with potential biological and toxicological activities, yet the active components elude definition. Ingestion of Aloe vera is associated with diarrhea, electrolyte imbalance, kidney dysfunction, and conventional drug interactions; episodes of contact dermatitis, erythema, and phototoxicity have been reported from topical applications. This review examines the botany, physical and chemical properties, and biological activities of the Aloe vera plant. PMID- 16690539 TI - Review of the evidence regarding the carcinogenicity of hexavalent chromium in drinking water. AB - Recent analyses have revealed that 38% of municipal sources of drinking water in California have detectable levels of hexavalent chromium. This observation provided new impetus to characterize the carcinogenic risk associated with oral exposure to hexavalent chromium in drinking water. Notwithstanding the well characterized increases in cancer associated with inhalation exposure to this chemical, the marked reduction of hexavalent chromium to trivalent chromium in the stomach suggests that exposure to hexavalent chromium in drinking water may not pose a carcinogenic risk. A reevaluation of studies that investigated the toxicokinetics, the genotoxicity, and the mechanism of carcinogenicity of hexavalent chromium, as well as the available human and animal cancer studies, was undertaken to determine if there is evidence that exposure to this chemical in drinking water may pose a carcinogenic risk. Mechanistic studies suggest the potential for a carcinogenic response if hexavalent chromium enters cells. Both toxicokinetic and genotoxicity studies indicate that a portion of an orally administered dose of hexavalent chromium is absorbed and gets into cells of several tissues, causing DNA damage. The only lifetime oral study of hexavalent chromium in animals conducted thus far yielded a statistically significant increase in stomach tumors compared to controls. Also, in a limited-term cancer study, co-exposure to hexavalent chromium in drinking water and ultraviolet light produced skin tumors in mice. The only available cancer study of humans exposed to hexavalent chromium in drinking water revealed a statistically significant increase in stomach tumors. Moreover, a meta-analysis of occupational studies also revealed a statistically significant increase in stomach cancers. The increases in stomach tumors in both human and animal studies, along with the toxicokinetic, genotoxic, and mechanistic data, suggest that oral exposure to this agent appears to pose a carcinogenic risk. PMID- 16690543 TI - Sensitivity of the Halstead and Wechsler Test Batteries to brain damage: Evidence from Reitan's original validation sample. AB - The Halstead-Reitan Battery has been instrumental in the development of neuropsychological practice in the United States. Although Reitan administered both the Wechsler-Bellevue Intelligence Scale and Halstead's test battery when evaluating Halstead's theory of biologic intelligence, the relative sensitivity of each test battery to brain damage continues to be an area of controversy. Because Reitan did not perform direct parametric analysis to contrast group performances, we reanalyze Reitan's original validation data from both Halstead (Reitan, 1955) and Wechsler batteries (Reitan, 1959a) and calculate effect sizes and probability levels using traditional parametric approaches. Eight of the 10 tests comprising Halstead's original Impairment Index, as well as the Impairment Index itself, statistically differentiated patients with unequivocal brain damage from controls. In addition, 13 of 14 Wechsler measures including Full-Scale IQ also differed statistically between groups (Brain Damage Full-Scale IQ = 96.2; Control Group Full Scale IQ = 112.6). We suggest that differences in the statistical properties of each battery (e.g., raw scores vs. standardized scores) likely contribute to classification characteristics including test sensitivity and specificity. PMID- 16690544 TI - Does impaired executive functioning differentially impact verbal memory measures in older adults with suspected dementia? AB - The purpose of this study was to examine whether executive dysfunction differentially impacts list-learning and story recall tasks in a sample of older adults referred for suspected cognitive impairment. Older adults (N = 61) with mild cognitive impairment (MCI) or probable mild dementia, and those who did not meet criteria for diagnosis of dementia, were assessed using measures of executive function and verbal memory. Two groups were established based on performance on measures of executive function: (a) the No Executive Dysfunction group (NoED; n = 33) consisted of persons without impairment on any obtained measures of executive function; and (b) the Executive Dysfunction group (ED; n = 28) contained persons with impairment on at least one of the measures of executive function. The two groups were compared on performance on two measures of verbal memory, the California Verbal Learning Test-II (CVLT-II) and the Logical Memory (LM) subtest from the Wechsler Memory Scale-Revised (WMS-R). The NoED group performed significantly better than the ED group on the total learning and short delay free recall trials of the CVLT-II. However, there were no significant differences between the groups on the other indices of the CVLT-II (i.e., long delay free recall, recognition, recall repetitions, recall intrusions, or recognition false-positives) or on the immediate and delayed recall trials of the LM measure. These results support previous research demonstrating the impact of executive dysfunction on the acquisition of and short delay retrieval of verbal information in older adults with suspected cognitive impairment. PMID- 16690545 TI - Early environmental factors, ethnicity, and adult cognitive test performance. AB - This study was conducted to better understand cross-cultural variation in neuropsychological test performance by exploring the relationship between early environmental factors and adult neuropsychological functioning in African Americans (AA) and Caucasians (CA). Cognitive testing data and detailed retrospective early environmental histories were obtained from 100 neurologically healthy adults (75 AA, 25 CA). Results indicate that: (a) consistent with previous research, the two ethnic groups differed significantly on their cognitive test performance; (b) early environmental factors were significantly related to performance on cognitive tests; (c) the two ethnic groups demonstrated slight, but significant, differences in their early environmental histories; but (d) covarying early environmental factors did not substantially reduce the group performance in cognitive test performance. The failure of reported environmental factors to account for more of the ethnic disparity in test performance may have resulted from the inherent weakness of the retrospective assessment method (i.e., lack of precision or objectivity in participants' recollections of their early environments). These results highlight the remaining need for ethnicity specific normative data until cross-cultural variation in neuropsychological test performance is better understood. PMID- 16690546 TI - Test-retest norms and reliable change indices for the MicroCog Battery in a healthy community population over 50 years of age. AB - The increasing availability of computerized test batteries used to assess neuropsychological changes requires the availability of suitable test-retest normative data. Reliable change indices can then be used to evaluate significance of change in an individual's performance on retesting. We tested (N = 40) neurologically normal adults on three occasions (initially, two weeks, and three months) on the MicroCog: Assessment of Cognitive Functioning computerized testing instrument. Normative retest data are presented for two analytic techniques: the Reliable Change Index adjusted for practice and the Standardized Regression-Based technique. At two weeks, the correlation coefficients ranged from .49 to .84, with all scores demonstrating significant practice effects. At 3 months, coefficients ranged from .50 to .83, with all scores except Attention / Mental Control demonstrating significant practice compared to baseline. Regression equations were generated for all scores using age, sex, education level, and score at Time 1 as predictors. For all measures the only significant predictor was the Time 1 score. The reliable change indices and regression equations presented here can be used to determine the significance of change from predicted retest scores in a matched interventional cohort. PMID- 16690547 TI - Trail making test errors and executive function in schizophrenia and depression. AB - The Trail Making Test (TMT) frequently is used as a measure of executive cognitive function. However, traditional use of test completion time as the primary outcome score does not give the more detailed information on cognitive processes that analysis of test-taking errors may provide. The present study compared TMT performance of three groups: patients with schizophrenia, patients with major depression, and healthy control participants (n = 30 for each group). Three operationally defined error types were examined: (a) tracking, (b) perseverative, and (c) proximity. Although both patient groups were slower than the healthy control group, only the schizophrenia group made significantly more errors, particularly tracking errors, suggesting a greater degree of cognitive disorganization. Within-group analysis of a larger group of schizophrenia patients (n = 84) revealed that TMT time was most strongly associated with the Withdrawal-Retardation factor of the Brief Psychiatric Rating scale. In contrast, TMT errors were most strongly associated with the Conceptual Disorganization factor. Comparisons of TMT scores and other cognitive tests showed moderate to high associations with tests of working memory, psychomotor speed, and executive function. Stepwise regression analysis revealed an independent association between Digit Cancellation and Part B Time, indicating a unique contribution of visuomotor scanning to performance. In contrast, Part B errors were uniquely associated with the Verbal Series Attention Test and the Token Test, tests of mental tracking and executive-mediated working memory, respectively. These findings demonstrate the utility of TMT error analysis in revealing cognitive deficits not traditionally captured using completion time as the sole outcome variable. PMID- 16690548 TI - Known-groups cross-validation of the letter memory test in a compensation-seeking mixed neurologic sample. AB - Compensation-seeking neuropsychological evaluees were classified into Honest (HON; n = 37) or Probable Cognitive Feigning (PCF; n = 53) groups based on results from the Victoria Symptom Validity Test, the Test of Memory Malingering, and the Digit Span subtest of the Wechsler Adult Intelligence Scale--3rd ed. The groups were generally comparable on demographic, background, and injury severity characteristics, although HON TBI participants were significantly more likely to have a documented loss of consciousness, whereas PCF participants were significantly more likely to be currently on disability. PCF participants scored significantly lower on many neuropsychological test, particularly of memory, as well as higher on most MMPI-2 clinical scales. The PCF group also had significantly higher scores on multiple indices of feigning of psychiatric symptoms. Results from the Letter Memory Test (LMT) were significantly lower for the PCF group, and using the recommended cutting score, specificity was .984, whereas sensitivity was .640, suggesting adequate performance on cross validation. PMID- 16690549 TI - Investigation of biases in the general public in evaluating mild head injury using neuropsychological and CT scan results: forensic implications. AB - The purpose of this study was to determine how the general public considers neuropsychological (NP) vs. CT scan data in assessing the effects of a possible mild traumatic brain injury (MTBI). Ten scenarios depicting an MTBI from a motor vehicle accident were presented to community volunteer participants. Case scenarios differed with regard to diagnostic information presented (i.e., all combinations of normal, abnormal, or absent NP and CT results were provided). The number of participants ranged from 31 to 42 across the 10 cases and there were no age or educational differences among the groups. Participants rated the likelihood of symptoms being attributable to brain damage from the accident on a 7-point Likert-type scale. Scenarios with positive NP or CT results were rated as more likely to have sustained brain damage than when NP/CT results were negative. However, when NP and CT data were contradictory (i.e., one positive and the other negative), there were no differences in the ratings of the participants. Thus, members of the general public did not demonstrate any preconceived bias about the validity of NP vs. CT results in MTBI. Women were more likely than men to attribute symptoms to brain damage from the motor vehicle accident, as did participants with a history of TBI. Forensic implications are discussed. PMID- 16690550 TI - The word reading test of effort in adult learning disability: a simulation study. AB - The Word Reading Test (WRT) was designed to detect effort problems specific to a learning disability sample. The WRT and the Word Memory Test (WMT) were administered to two simulator and normal control groups. The WRT showed excellent receiver operating characteristics (e.g., 90% sensitivity and 100% positive predictive power) and outperformed the WMT in detecting both reading and mental speed simulators. This finding and a double dissociation between reading and speed simulators on WRT errors and reaction time suggested specific effort effects while poor effort of simulators on the WMT suggested general effort effects. Results are supportive of the WRT as a potential effort indicator in learning disability. PMID- 16690551 TI - Doing research on medical education. PMID- 16690552 TI - Good news from Denmark! PMID- 16690553 TI - Exercise on prescription in general practice: a systematic review. AB - 'Exercise on Prescription' (EoP) is used for initiating physical activity among sedentary patients with signs of lifestyle diseases. EoP is personalized secondary prevention in primary healthcare. This review addresses EoP using a Health Technology Assessment perspective and aims to answer the following questions: (1) Does EoP increase physical activity level or physical fitness, and is more intensive EoP more effective than less intensive? (2) Is EoP acceptable and feasible in general practice? (3) Is EoP acceptable to and feasible for sedentary patients? (4) Is EoP cost-effective? EoP studies were searched using Medline thesaurus topic, Medline WinSPIRS, reference lists of recent reviews, and NLM Gateway Locator plus. A total of 22 studies were included in the review. Most studies reported moderate improvements in physical activity or physical fitness for 6-12 months. Among patients receiving EoP 10% more had improved physical activity level compared with controls and mean aerobic fitness was improved by 5 10% among EoP patients compared with controls. Little evidence existed in support of the hypothesis that more intensive EoP is more effective. EoP was acceptable and feasible to GPs and patients volunteering for EoP. However, little is known about non-completers, patients declining EoP, and GPs not highly motivated for using EoP. Only one study addressed health economic issues. It found EoP cost effective, but comparisons with other interventions have not been performed. Even though most studies in this review presented favourable results for EoP there is a lack of evidence in several fields. In particular we lack high-quality studies evaluating EoP schemes that are sustainable in everyday use in general practice. PMID- 16690554 TI - Smoking cessation in patients with diabetes mellitus: results from a controlled study of an intervention programme in primary healthcare in Sweden. AB - OBJECTIVE: To evaluate an intervention programme on smoking cessation in patients with diabetes mellitus in primary healthcare. DESIGN: Regional controlled intervention study. SETTING: Seventeen primary healthcare centres in Sweden. INTERVENTION: In the intervention centres, nurses with education in diabetes were given one half-day of training in motivational interviewing and smoking cessation. An invitation to participate in a smoking cessation group was mailed to patients from the intervention centres followed by a telephone call from the patient's diabetes nurse. The nurses who intervened were specially educated in smoking cessation. The control group received a letter containing advice to stop smoking and information about a one-year follow-up. PATIENTS: Daily smokers with diabetes mellitus, 30-75 years of age. In the intervention centres 241 patients fulfilled the criteria and in the control centres 171 patients. MAIN OUTCOME MEASURES: Self-reported smoking habits after one year. RESULTS: In total, 21% of the smokers accepted group treatment. After 12 months, 20% (42/211) in the intervention centres reported that they had stopped smoking and 7% (10/140) in the control centres; 40% (19/47) of the smokers who had participated in group treatment reported that they had stopped smoking. CONCLUSION: A computerized record system for patients in primary healthcare was used to identify diabetic patients who were smokers. The selected group was invited to a stop smoking programme. At a one-year follow-up significantly more patients in the intervention centres had stopped smoking compared with patients in the control centres. PMID- 16690555 TI - Variation in spirometry utilization between trained general practitioners in practices equipped with a spirometer. AB - OBJECTIVE: To explore spirometry utilization among general practitioners and identify practitioner and practice-related factors associated with spirometry utilization. DESIGN: Multivariate multilevel cross-sectional analysis of a questionnaire survey. SETTING: Some 61 general practices involved in a spirometry evaluation programme in the Netherlands. All practices owned a spirometer and were trained to perform spirometry. SUBJECTS: A total of 144 general practitioners and 179 practice assistants. MAIN OUTCOME MEASURES: Extent of spirometry utilization for five indications from national COPD/asthma guidelines, practitioner and practice-related factors associated with spirometry utilization. RESULTS: The response rate was 97%. General practitioners used spirometry mostly to evaluate treatment with inhaled steroids (58%). Significant practitioner related factors associated with spirometry utilization were: general practitioners' job satisfaction, general practitioners' general interest in research, and prior participation in spirometry training. Practice-related factors associated with spirometry utilization were: presence of a practice nurse, delegation of medical tasks to practice assistants, use of spirometry in different rooms, and use of protocols in practice. CONCLUSION: Practitioner- as well as practice-related factors were associated with the extent of spirometry utilization. In particular, it is essential to improve practice-related factors (e.g. presence of a practice nurse, more delegation of medical tasks to the practice assistant). PMID- 16690556 TI - Detection of middle-ear fluid in children with spectral gradient acoustic reflectometry: a screening tool for nurses? AB - OBJECTIVE: To evaluate the validity of spectral gradient acoustic reflectometry (SG-AR) in the hands of nurses in screening children for middle-ear fluid (MEF). DESIGN: Prospective, blinded study. SETTING: A satellite study within the Finnish Otitis Media Vaccine Trial in primary care in 1995-99. PATIENTS: Some 739 ear examinations among a cohort of 271 children under the age of 2 years during different healthcare contacts (acute sick visits, check-up visits after otitis media, and scheduled healthy control visits at 24 months of age). MAIN OUTCOME MEASURES: Specificity, sensitivity and positive and negative predictive values of SG-AR performed by nurses in detecting MEF using pneumatic otoscopy by trained physicians as a reference. RESULTS: SG-AR was successful in 585 (79%) ears. None of the cut-off points assessed resulted in both excellent sensitivity and specificity. Nevertheless, at the sick visits, positive predictive values at 50 and 60 degrees were as high as 94% and 89%, respectively. However, negative predictive value for the cut-off point of 100 degrees was considerably lower, at 75%. At the non-acute visits, negative predictive values were excellent. CONCLUSION: SG-AR is a useful device for nurses in screening MEF among children. It detects ears with both very high and very low probability of MEF and is especially effective in detecting MEF during sick visits and in ruling out MEF among non-acute patients. We recommend the use of the SG-AR cut-off point of 60 degrees as a sign of MEF when screening sick children, and the cut-off point of 100 degrees as a sign of a healthy ear among non-symptomatic patients. PMID- 16690557 TI - Clinical course of suspected viral sore throat in young adults: cohort study. AB - OBJECTIVE: To evaluate the natural clinical course of suspected viral sore throat. DESIGN: Prospective cohort study. SETTING: Five military camps in Norway. METHODS: In a randomized placebo-controlled trial comparing the effect of bovine colostrum tablets or placebo on non-streptococcal sore throat in young adults no statistically significant difference in illness duration was found. The participants were thus regarded as one cohort. The authors analysed 10.0 cm visual analogue scales (VAS) scores for 'sickness' and sore throat, using 1.0 cm as a cut-off for recovery. They furthermore explored whether the VAS for sore throat was a good test to discriminate between recovered/not recovered by use of a receiver operating characteristic (ROC) curve. RESULTS: For sore throat, it was found that 51% had recovered by day 6 and 91% had recovered by day 8. Similarly for sickness, 65% had recovered by day 6 and 94% by day 8. The daily VAS scores for 'sickness' and sore throat were highly correlated (p = 0.001). The mean day of recovery from 'sickness' and sore throats (based on VAS scores) was 5.5 and 4.7 respectively and 5.3 based on the dichotomous outcome for throat pain. The ROC curve revealed that a VAS score < 1.0 was the best cut-off point for recovery, with a sensitivity of 0.74 and a specificity of 0.90. CONCLUSION: Almost all the patients had recovered by day 8. A high correlation was found between sickness and sore throat. These data will assist clinicians and others promoting self-care to set realistic expectations regarding recovery from URTI in young adults. PMID- 16690558 TI - Infections in families with small children: use of social insurance and healthcare. AB - OBJECTIVE: To examine infectious symptoms on a daily basis in families with small children and how often these infections cause people to stay at home or seek healthcare. DESIGN: A population-based prospective study. SETTING: Child health clinics in seven municipalities in Sweden. SUBJECTS AND MAIN OUTCOME MEASURES: All family members of 835 families who came with an 18-month-old child to a child health clinic were asked to register all infectious symptoms in a diary for a month. They were also asked to indicate whether they had stayed at home from day care or school, whether social insurance had been used, and whether they had contacted healthcare facilities or seen a physician. RESULTS: In total, 7% of the 18-month-old children and 34% of the parents had no symptoms during the winter month. The most common symptom was a runny nose. The 18-month-old children had 1.6 symptom episodes with an average duration of 5.6 days. Of the symptom episodes 13% led to contact with healthcare facilities and 6% to an antibiotic prescription. Of the symptom days 27% required staying at home and in 10% social insurance was claimed. CONCLUSION: Symptoms of infection among families with small children were common, with a runny nose being the most common. Physician consultations and antibiotic prescriptions were used in a small proportion of the symptom episodes. Social insurance was claimed in about one-third of the days with absence from day-care. PMID- 16690559 TI - Impact of physician-related factors on sickness certification in primary health care. AB - OBJECTIVE: To investigate what factors are associated with physicians' decisions on whether or not to sickness certify the patient at a consultation. DESIGN: Questionnaire survey sent to physicians in general practice and their patients. SETTING: General practitioners in Orebro county, central Sweden. SUBJECTS: Sixty five physicians with up to 10 patients each. MAIN OUTCOME MEASURE: Whether a sickness certificate was issued. RESULTS: Physicians with long experience in family medicine and those working part time issued more sickness certificates when all encounters with patients were considered. When only musculoskeletal problems were studied physicians with long experience or who were trained in social insurance medicine as undergraduates issued more sickness certificates. When only appointments for infections were studied, part-time physicians issued more sickness certificates. No impact of the physicians' sex on sickness certifying was found. CONCLUSIONS: Length of professional experience and physicians' working time appear to influence practices in sickness certifying. PMID- 16690560 TI - Concordance between elderly patients' understanding of and their primary healthcare physician's diagnosis of heart failure. AB - OBJECTIVE: The objective was to study primary healthcare patients' understanding of their diagnosis of heart failure (HF), using patients treated for diabetes mellitus (DM) as a comparative group. DESIGN: A cross-sectional community based study. SETTING: Karlskrona community situated on the Swedish south-east coast with 60,600 inhabitants. SUBJECTS: A total of 1402 subjects, aged 60-96 years in 10 age cohorts, selected randomly from the national population registry participating in the Swedish National study on Ageing and Care - Blekinge. MAIN OUTCOME MEASURES: Understanding of diagnosis of HF or DM in primary healthcare. Prevalence of cognitive impairment. RESULTS: In all, 39.8% of patients with a diagnosis of HF treated in primary healthcare and 97.1% of patients with DM had an understanding of their respective diagnosis. Cognitive impairment was significantly more prevalent in the groups of patients treated for HF (OR 1.9, 95% CI 1.2 to 3.0) and DM (OR 1.8, 95% CI 1.1 to 3.1), when compared with those not treated for either HF or DM. The odds ratio for understanding of diagnosis was 0.013 (95% CI 0.003 to 0.052, p < 0.001) in patients treated for HF, compared with patients treated for DM, when adjusted for the subject's age, sex, and cognitive function. CONCLUSION: In this study it was shown that patients' understanding of their diagnosis was highly dependent on diagnosis, independently of age, sex, or cognitive function. The results suggest that there is room for improvement of care in primary healthcare, to increase HF patients' understanding of their diagnosis. PMID- 16690561 TI - Why do sales of lipid-lowering drugs vary between counties in Norway? Evidence from the OPPHED Health Study 2000-2001. AB - OBJECTIVE: To study and compare plausible factors that might explain varying sales of lipid-lowering drugs (LLDs) in the two neighbouring counties of Hedmark and Oppland in Norway, with a similar age distribution, socioeconomic structure, and access to healthcare services. DESIGN, SETTING, SUBJECTS: Cross-sectional population study comprising 10 598 attendants aged 40, 45, 60, and 75 years in the OPPHED Health Study, 2000-2001 (attendance rate 61%). MAIN OUTCOME MEASURE: Treatment eligibility (cardiovascular morbidity and risk score), treatment frequency in treatment-eligible subgroups and treatment intensity in terms of achievement of total cholesterol (TC) goal. RESULTS: Proportions eligible for LLD treatment in Hedmark and Oppland were similar. There was no difference in prevalence of LLD use among participants with cardiovascular disease or diabetes (secondary prevention subgroup). However, LLD use among men in the primary prevention subgroup was higher in Hedmark compared with Oppland, 6.3% and 4.1%, respectively (p < 0.05). The same tendency was seen among women. In both sexes, more LLD users in the primary prevention subgroup achieved the TC goal in Hedmark compared with Oppland (p < 0.05). CONCLUSION AND IMPLICATIONS: The proportion of the population eligible for LLD treatment in the two counties should imply similar treatment rates in both. Higher LLD treatment frequency and intensity in the primary prevention subgroup in Hedmark are probably both contributing factors that explain the higher sales of LLDs in Hedmark compared with Oppland. Feasible intervention thresholds for primary prevention with concurrent reimbursement rules should be defined in guidelines to avoid unintentional variation in LLD use in the future. PMID- 16690562 TI - Working behaviour of competent general practitioners: personal styles and deliberate strategies. AB - OBJECTIVE: To study how competent GPs perform their work within the consultation and in relation to the team and practice organization. DESIGN: Ethnographic study with thick description. A participant observation of the GP was followed by a personal interview. A substantial description was elaborated that was analysed qualitatively. SETTING: Primary care surgeries in Sweden. SUBJECTS: A number of competent GPs. RESULTS: Two main categories emerged, deliberated strategy and personal style. These categories set up the individual pattern of working behaviour for each GP. A behaviour that is a deliberate strategy for one GP for the other belongs to his or her personal style. Common denominators in the overall working behaviour were attention to the patient as a person, practising patient-centred medicine, saving the consultations from disturbances, rejecting taking over responsibilities from the patients, and safeguarding own autonomy. CONCLUSION: The transition of deliberate, favourable strategies into one's personal style is an important aspect of professional development. A well developed personal style is necessary to obtain the spontaneous interchange between attentive listening and detachment characteristic of patient centeredness. PMID- 16690563 TI - Gait parameters as predictors of slip severity in younger and older adults. AB - This study investigated the association between slip severity and pre-slip gait characteristics of younger and older subjects. Sixteen younger and eleven older healthy adults walked onto an unexpectedly slippery surface. Slip severity was categorized as either hazardous or non-hazardous using a 1.0 ms peak slip velocity threshold. The results showed that hazardous slips were associated with greater step lengths (normalized by leg length) (SLR), larger and more rapidly changing foot - floor angles (FFA) at heel strike, and increased cadence across the two subject groups. Older subjects were found to walk with shorter SLR and with smaller and more slowly changing FFA at heel strike compared to younger subjects. However, both younger and older subjects experienced hazardous slips at the same rate. A logistic regression model relating SLR and cadence to slip severity predicted that increased SLR and decreased cadence would result in increased probability of hazardous slip (R2 = 0.45, chi2 = 15.30, p<0.01). A second logistic regression model relating FFA with slip severity predicted that increased FFA would result in increased probability of hazardous slip (R2 = 0.53, chi2 = 16.55, p<0.01). These results suggest that gait characteristics prior to foot contact play an important role in the severity of an ensuing slip. The finding that older adults experienced hazardous slips at the same rate as young adults even though their SLR and FFA are smaller suggests that age is also playing a role in other aspects of postural control that impact slip severity. PMID- 16690564 TI - Muscle fatigue and endurance during repetitive intermittent static efforts: development of prediction models. AB - Localized muscle fatigue has received growing attention as a potential design variable and exposure metric in research towards prevention of musculoskeletal disorders in the workplace. While fatigue during sustained static work has been investigated extensively, effects during tasks comprising work-rest cycles are less clear. Work-rest models for static intermittent work have been presented in several reports, but the applicability is often limited to specific conditions. A study was conducted that facilitated a description of the relationships between static intermittent efforts and muscle endurance and fatigue. Exercises consisted of 1 h (maximum) of repetitive static arm abductions, involving a range of muscle contraction levels (10-30% maximum voluntary exertion), duty cycles (0.2-0.8) and cycle times (20-180 s). A between-subject central composite experimental design was used and 15 different exercise conditions were examined with six participants (three females and three males) for each. Along with endurance times, temporal changes related to fatigue were monitored using muscle strength, ratings of discomfort and electromyography (EMG) obtained from the middle-deltoid muscle during the contraction phase of the work cycles. The results of this study showed the influence of contraction level and duty cycle on the majority of fatigue measures used, while cycle time tended to affect EMG spectral measures. Using a response surface methodology, several fatigue prediction models and contour plots were developed that can be employed as an aid for design and evaluation of light repetitive static tasks. Good correspondence was generally found between discomfort rating and other measures of fatigue, suggesting the usefulness of this measure for rapid assessments of local fatigue in the workplace. PMID- 16690565 TI - Quantifying repetitive hand activity for epidemiological research on musculoskeletal disorders--part I: individual exposure assessment. AB - An exposure measurement approach is described for quantifying repetitive hand activity of individual workers in a prospective epidemiological study on work related upper extremity musculoskeletal disorders. A total of 733 subjects were involved in this study at the baseline. Hand activities were quantified by force and repetition. Force levels were measured by workers' self-reports, ergonomists' estimates based on observation and measurements with instrumentation. Repetition levels were measured by detailed time-motion analyses using two repetitive hand activity definitions and ergonomists' estimates using scales for the American Conference of Governmental Industrial Hygienists hand activity level and the Strain Index. Results showed that the present exposure assessment approach seems to be able to quantify force level and repetitiveness of hand activities. Repetitive hand activity is quantified differently depending on whether forceful hand exertion or repetitive muscle activity is used as the definition. These hand activity definitions may quantify different physical exposure phenomena. Individual exposure assessment is important in epidemiological research of musculoskeletal disorders as there are interactions between the individual subjects and the measured parameters. These interactions may vary between exposure parameters. PMID- 16690566 TI - Quantifying repetitive hand activity for epidemiological research on musculoskeletal disorders--part II: comparison of different methods of measuring force level and repetitiveness. AB - This paper focuses on comparisons between the different methods of assessing repetitive hand activities. Various methods were used to measure hand force and repetitiveness of hand activities on 733 subjects in the study described by Bao et al. (2006). Two definitions of repetitiveness were used in analysis of detailed time studies of repetitive hand activities and four parameters of the American Conference of Governmental Industrial Hygienists (ACGIH) hand activity level (HAL) and the Strain Index methods were estimated by ergonomists and used to quantify repetitiveness. Hand forces were measured or estimated using three different methods: 1) measured with a force gauge or mimicked on a force gauge (force matching); 2) estimated by ergonomists using rating scales; 3) self reports by subjects. The jobs were also evaluated using the ACGIH HAL and Strain Index methods when different repetitiveness quantification methods were used. Results showed that different definitions of repetitive exertion might lead to measuring different physical exposure phenomena and produce very different results. There were poor correlations between the measures of repetitiveness estimated by the different methods. Correlations between force quantifications using different methods were also poor. This suggests that parameters measured by different methods might not be interchangeable. Both the ACGIH HAL and Strain Index methods identified more 'hazardous' jobs when repetitiveness was estimated by ergonomists than when it was calculated by detailed time studies of forceful hand exertions. The Strain Index method identified more 'hazardous' jobs than the ACGIH HAL method. Overall, the between-methods agreements were found to be moderate to substantial. PMID- 16690567 TI - Observational assessment of situation awareness, team differences and training implications. AB - The two goals were to investigate, first, the practicability and reliability of observational assessment of team situation awareness (SA) and, second, the nature of any team differences, their consistency and training implications. Five shift teams tackled three scenarios, each with three probe events concerning SA, and three observers viewed and rated concurrently each shift. This methodology was found to be practicable and achieved satisfactory rater reliability as indicated by intraclass and inter-rater correlations. Team differences in SA emerged although there was no consistent pattern. A retrospective analysis of individual and team behaviours relating to SA was performed using the Critical Incident Technique. A total of 75 incidents and 20 behavioural dimensions relevant to SA were identified and these were subsumed under planning, problem solving, team coordination, attention, communication and knowledge. These findings are discussed with regard to the nature and measurement of SA, and the content of training to improve SA for control room teams. PMID- 16690568 TI - The impact of various rehydration volumes for firefighters wearing protective clothing in warm environments. AB - This study examined different fluid replacement quantities during intermittent work while wearing firefighting protective clothing and self-contained breathing apparatus in the heat (35 degrees C, 50% relative humidity). Twelve firefighters walked at 4.5 km per h with 0% elevation on an intermittent work (50 min) and rest (30 min) schedule until they reached a rectal temperature of 39.5 degrees C during work periods and 40.0 degrees C during rest, heart rates of 95% of maximum and/or exhaustion. During the heat-stress trials subjects received one of four fluid replacement quantities, high (H), moderate (M), low (L), and no hydration (NH), where H, M and L represented 78%, 63% and 37% of fluid loss, respectively. The total tolerance time (work + rest) was significantly greater during H (111.8 +/- 3.5), M (112.9 +/- 5.2) and L (104.2 +/- 5.8) compared to NH (95.3 +/- 3.8). In addition, work time (min), which excluded rest periods, was significantly greater in H (82.6 +/- 3.5), and M (82.9 +/- 5.2) compared to NH (65.3 +/- 3.8). It is concluded that incorporating even partial fluid replacement strategies while wearing firefighting protective clothing and self-contained breathing apparatus in the heat improves tolerance time. PMID- 16690571 TI - Skin problems in lower limb amputees: a systematic review. AB - PURPOSE: Skin problems of the stump in lower limb amputees are relative common in daily rehabilitation practice, possibly impeding prosthetic use. This impediment may have great impact in daily life. Our objective was to review literature systematically concerning incidence and prevalence of skin disorders of the stump in lower limb amputees. METHOD: A literature search was performed in several medical databases (MEDLINE, CINAHL, EMBASE, RECAL) using database specific search strategies. Reference lists in the identified publications were used as threads for retrieving more publications missed in the searches. Only clinical studies and patient surveys were eligible for further assessment. RESULTS: 545 publications were initially found. After selection, 28 publications were assessed for research methodology. Only one publication fulfilled the selection criteria. The prevalence of skin problems in a series of 45 lower leg amputees of 65 years and older was 16%. CONCLUSIONS: Prevalence and incidence of skin problems of the stump in lower limb amputees are mainly unknown. PMID- 16690572 TI - Comparison of the Orpington Prognostic Scale (OPS) and the National Institutes of Health Stroke Scale (NIHSS) for the prediction of the functional status of patients with stroke. AB - PURPOSE: The aim of our study is to compare the Orpington Prognostic Scale (OPS) and the National Institutes of Health Stroke Scale (NIHSS) and to evaluate whether they help us estimate the future functional status of patients with stroke. METHOD: Twenty-five patients with stroke were administered the OPS and NIHSS on the 7th day of stroke in order to define the severity of the disease, and the Barthel Index was performed in order to evaluate the functional status and the activities of daily living (ADL) at the 1st, 3rd, and 6th months. RESULTS: Both scales were statistically correlated (P = 0.0001). When the predictability of these scales in terms of the ADL and functional status was evaluated, the regression coefficient at the 1st month was -14.746, R(2) = 0.58, P < 0.0001 and -4.885, R(2) = 0.50, P < 0.0001 for OPS and NIHSS, respectively, the same coefficient at the 3rd month was -12.482, R(2) = 0.41, P = 0.001 for OPS and -3.280, R(2) = 0.23, P = 0.016 for NIHSS, and at the 6th month it was 11.662, R(2) = 0.38, P = 0.001 for OPS and -2.997, R(2) = 0.20, P = 0.02 for NIHSS. CONCLUSION: In patients with stroke, OPS and NIHSS had significant contribution to the estimation of the functional status and OPS was more effective than NIHSS. PMID- 16690573 TI - Predictors of return to work following traumatic work-related lower extremity amputation. AB - PURPOSE: To determine factors predictive of return to work (RTW) and days of total disability (TD) in a population of persons working at the time of lower extremity amputation. METHOD: Retrospective chart and database review. RESULTS: Of 88 valid cases, 48% involved toe amputation, 23% transtibial, 14% partial foot, 14% transfemoral, and 2% high level. Fifty-eight percent of all subjects RTW, 19% were deemed 'fit for work', and 23% did not RTW. Days TD ranged from 0 to 1664, with a mean of 366 days. Toe amputation level showed a mean of 127 days of TD. Bivariate analysis showed amputation level, total costs to Workers Compensation Board (WCB), and days TD significantly related to RTW, and rehabilitation costs, vocational rehabilitation, work assessment, age, number of surgical procedures, number of days in acute care, and amputation level significantly related to days TD. In the multivariate model, only amputation level and higher gross annual income showed predictive value for RTW. However older age, more surgical procedures, less days in hospital, and higher amputation levels were all predictive of increased days TD. CONCLUSION: Toe amputation level had a surprisingly high number of days TD, which may have significant potential economic and disability impact on the workplace. Other factors beyond simply amputation level (such as previous income level) are important considerations for RTW. PMID- 16690574 TI - Gambling problems and health functioning in individuals receiving disability. AB - PURPOSE: This study evaluated the rates and correlates of disordered gambling, with a focus on gambling behavior among participants receiving disability. METHOD: The sample consisted of 723 patients seeking free or reduced-cost dental care. Participants completed the South Oaks Gambling Screen (SOGS), Short Form-12 Health Survey, Second Edition (SF-12v2), and questions assessing demographic characteristics and frequency and intensity of current gambling behaviors. RESULTS: Results indicate a significantly higher prevalence of disordered gambling among participants receiving disability (26%) than in the remainder of the sample (14%; p < 0.001). Of the 135 individuals receiving disability, disordered gambling was associated with decreased physical and mental health functioning. CONCLUSIONS: These data indicate that individuals receiving disability have high rates of disordered gambling, and gambling behavior in this population is associated with poorer health functioning. Results suggest that disordered gamblers who receive disability have an increased need for interventions to reduce gambling and associated problems. PMID- 16690575 TI - Comparison of three definitions of the mid-stance and mid-swing events of the gait cycle in children. AB - PURPOSE: Different criteria are used to define the terms mid-stance (MST) and mid swing (MSW) when describing the gait cycle. None of these definitions is universally accepted. This causes difficulties with the interpretation of gait analysis data and hinders the comparison between the different studies. The aim of the present study was to compare three definitions of MST and MSW by examining the gait of a group of healthy children. METHOD: A prospective comparison of three commonly used definitions of the MST and MSW events of the gait cycle. OUTCOME MEASURES: The timing of the temporal, kinematic and kinetic MST and MSW of the gait cycle. RESULTS: Thirty healthy 8 - 10-year-old children were studied. There was no consistent correlation between the timing of the chosen definitions of MST and MSW. However, there was a tendency for the timing of the temporal and kinetic MST and MSW to occur, respectively, at approximately 30 and 80% of the gait cycle. CONCLUSIONS: The temporal definition of MST and MSW as the midpoints in time of the respective phases of the gait cycle appears to be more appropriate than other definitions. The use of this definition may prevent misunderstanding and permit comparison of the results of the different gait analysis studies. PMID- 16690576 TI - Modelling 5-year functional outcome in a major traumatic injury survivor cohort. AB - AIM: To estimate associations between possible predictors of functional disability outcome at 5 years in a working adult population cohort of survivors of major traumatic injury and to develop a prognostic model of outcome. DESIGN: Population based retrospective cohort study. POPULATION: Persons who had experienced major traumatic injury (Injury Seventy Score >15) in the area of the former Yorkshire Regional Health Authority during the period 30 September 1988 to 1 October 1989 and who survived for 5 years (average 5.3 years). METHODS: The same interviewer saw each survivor at their home and used the OPCS Adult Disability Schedule to ascertain levels of functional disability. Disability scores for each survivor were then combined in accordance with the OPCS guidance to calculate a level of disability between 0 (no disability) to 10 (maximum disability). The OPCS level was then dichotomised with a cut-point at 4/5. Possible predictor and confounding variables from pre-injury, injury and post injury periods were modelled in a logistic regression to identify those that predicted outcome level. Two reduced models were developed to allow early prognosis of late outcome. RESULTS: The full model correctly classified 91% of observed outcomes. Inpatient length of stay OR 1.031 (95% CI 1.014,1.048) per day predicted poorer 5-year outcome; Glasgow Coma Score OR 0.790(0.629,0.992) per 1 point increase; stay in Intensive Care Unit OR 0.931 (0.877,0.987) per day; attainment of degree-level education OR 0.014 (0.000,0.707); single civil status OR 0.110 (0.013,0.908); being taken initially to a hospital with a neurosurgical facility OR 0.064(0.010,0.420); being in paid work during the 2 weeks before index injury OR 0.093(0.009,0.969) predicted better 5-year disability outcome. Two reduced models were constructed that included a simple set of variables, one of these models excluded any rehabilitation variables but still correctly classified 85% of the observed outcomes. CONCLUSION: As well as level of traumatic brain injury (TBI) and total inpatient stay, Pre-injury educational attainment and employment, civil status, immediate care in a hospital with a neurosurgical facility and stay in an Intensive Care Unit determined 5-year outcome. It is possible to efficiently predict outcome at an early stage. Previous work on predictors of disablement have suffered from large selection and attrition biases. PMID- 16690577 TI - Malnutrition, eating difficulties and feeding dependence in a stroke rehabilitation centre. AB - PURPOSE: To analyse prevalence of malnutrition, eating difficulties and feeding dependence in stroke rehabilitation patients because little is known about these prevalence's. Stroke patients have an increased risk for developing eating difficulties, feeding dependence and malnutrition because of their neurological deficits. METHODS: Malnutrition is defined by having unintentional weight loss or a low BMI. Structured observations of eating difficulties and feeding dependence were performed at a regular meal. RESULTS: During the study period of 18 months 69 patients of the 114 admitted stroke patients were included in our study. On admittance malnutrition was present in 35% of the patients and decreased to 3% after 4 weeks of rehabilitation. In total 43% of the patients had one or more eating difficulties and 16% was dependent in feeding. Aberrant eating speed was significantly associated with the presence of malnutrition at admission. CONCLUSIONS: The prevalence of malnutrition in stroke rehabilitation patients is high and decreases significantly during four weeks of rehabilitation. Research is needed into constructs of malnutrition, eating difficulties and feeding dependence. PMID- 16690578 TI - A preliminary economic evaluation of percutaneous neuromuscular electrical stimulation in the treatment of hemiplegic shoulder pain. AB - OBJECTIVE: The objective of this study was to compare the cost-effectiveness of various treatment modalities for hemiplegic shoulder pain. DESIGN: A stage II economic evaluation. MAIN OUTCOME MEASURES: Incremental cost effectiveness ratio of P-NMES, compared to slings and anti-inflammatory injections. RESULTS: The incremental cost effectiveness ratio (ICER) of p-NMES, compared to anti inflammatory injections is 6,061 euro(+/-3,285). The incremental cost of the first quality-adjusted life year after implantation of the P-NMES device compared to anti-inflammatory injections is 33,007 euro (+/-5,434). This decreases to approximately 7,000 euro after 5 years, and to approximately 5,000 euro after 10 survival years. CONCLUSION: In this early evaluation, P-NMES seems to be cost effective according to known guidelines. Treatment with P-NMES is recommended for patients with chronic HSP. PMID- 16690579 TI - Comparison of a self-report and performance-based test of disability in people with systemic lupus erythematosus. AB - PURPOSE: The purpose of this study was to examine functional ability in people with SLE by comparing a self-report and a performance-based test. METHOD: Fifteen women with SLE and 15 healthy controls participated in this study. Participants completed a self report of daily living skills, the Health Assessment Questionnaire (HAQ). Actual performance during activities of daily living was evaluated with the Assessment of Motor and Process Skills (AMPS). Measures of cognition and disease activity were also collected. RESULTS: HAQ scores of the participants with SLE indicated only very mild disability while scores on the AMPS indicated ineffective skill performance and potential safety risks. In the participants with SLE, cognitive status correlated significantly with the processing component (r(s) = 0.62, p < 0.05) and disease severity correlated with the motor component of the AMPS (r(s) = 0.84, p < 0.001). The HAQ did not correlate with the AMPS. CONCLUSIONS: This study suggests that cognitive deficits were more related to scores on the performance-based test of functional ability rather than the self-report. The performance-based test appeared to provide information that was not gained through self-report and measures of disease activity and gross cognition. PMID- 16690580 TI - Adjusting expectations after spinal cord injury across global settings: a commentary. AB - PURPOSE: The objective of this paper is to stimulate thought and discussion as to how best to set rehabilitation goals to maximize activities and participation of persons with spinal cord injury, across global settings where circumstances and environments may be widely different. METHOD: A review of literature and commentary are presented. Three points are articulated: (1) rehabilitation professionals need to understand factors that impact upon activities and participation, and need measurement tools that report these factors, in order to better appreciate outcomes in different settings, (2) rehabilitation professionals generally set goals with patients, but current measures of activities and participation do not indicate when or why maximal achievable function is sometimes not chosen, and (3) we need to develop realistic expectations for activities and participation after SCI in settings where current standard outcome chart targets are not feasible, due to socio-economic circumstances. CONCLUSIONS: A standardized approach to reporting measures of activities and participation, along with factors that influence these scores, is required for purposes of comparing rehabilitation outcomes in settings of differing socio-economic environments. In regards to spinal cord injury rehabilitation, an accepted standard of setting achievable rehabilitation goals is required for each level of complete spinal cord injury that could apply in various global settings. PMID- 16690581 TI - Using the ICF to clarify team roles and demonstrate clinical reasoning in stroke rehabilitation. AB - PURPOSE: The International Classification of Functioning, Disability and Health (ICF) is advocated as a tool to structure rehabilitation and a universal language to aid communication, within the multi-disciplinary team (MDT). The ICF may also facilitate clarification of team roles and clinical reasoning for intervention. This article aims to explore both factors in stroke rehabilitation. METHOD: Following a review of the literature, a summary was presented and discussed with clinicians working within stroke rehabilitation, to gather expert opinions. The discussions were informal, being part of service development and on-going education. The clinicians summarised key themes for the potential use of the ICF within clinical practice. RESULTS: Two key themes emerged from the literature and expert opinion for the potential use of the ICF in stroke rehabilitation: (i) to aid communication and structure service provision, (ii) to clarify team roles and aid clinical reasoning. Expert opinion was that clarification of team roles needs to occur at a local level due to the skill mix, particular interests, setting and staffing levels within individual teams. The ICF has the potential to demonstrate/facilitate clinical reasoning, especially when different MDT members are working on the same intervention. CONCLUSION: There is potential for the ICF to be used to clarify team roles and demonstrate clinical reasoning within stroke rehabilitation. Further experiential research is required to substantiate this view. PMID- 16690583 TI - Psychological well-being of adults with acquired hearing impairment. AB - AIM: To study psychological well-being (health-related quality of life) in a population of adults 20 years and over with hearing impairment (HI) and its relation to audiological factors, consequences of the HI, sense of humour, and use of communication strategies. SUBJECTS AND METHODS: Consecutive adults (n = 343) at the outpatient Unit of Audiology of a Norwegian university hospital answered the Psychological General Well-being inventory (PGWB), Hearing Disability and Handicap Scale (HDHS), Sense of Humour Questionnaire-6 (SHQ-6), and Communication Strategies Scale (CSS) in relation to an audiological examination and medical consultation. RESULTS: Mean PGWB index for the whole sample was 81.4 (SD 14.3) and females reported a significantly lower psychological well-being. In multiple linear regression analyses well-being was negatively associated with high levels of activity limitation and participation restriction. PGWB index was positively associated with high sense of humour, but was neither explained by audiological factors nor use of communication strategies. CONCLUSIONS: Psychological well-being was associated with the outcome of a standard HI assessment of activity limitation and participation restriction, but not with degree of HI and use of communication strategies. PMID- 16690584 TI - Prediction of long-term occupational performance outcomes for adults after moderate to severe traumatic brain injury. AB - PURPOSE: To examine predictors of long-term occupational performance outcomes for adults after moderate to severe traumatic brain injury (TBI). METHOD: This study involved analysis of data from a retrospective cohort of adults (N = 306) with moderate to severe TBI discharged from a Pennsylvania rehabilitation treatment facility. Extensive pre-injury sociodemographic, injury-severity, post-injury personal (cognitive, physical, affective), post-injury environmental (social, institutional, physical), and post-injury occupational performance (participation in self-care, productivity, leisure activities) data were gathered from hospital records and using in-person interviews. Interviews occurred at a mean time of 14 (range, 7-24) years post-injury. Hierarchical multiple regression analysis was used to investigate determinants of long-term occupational performance outcomes. RESULTS: Pre-injury behavioural problems, male gender, post-injury cognitive and physical deficits, and lack of access to transportation were significant independent predictors of worse occupational performance outcomes. CONCLUSIONS: The study supports the use of a comprehensive model for long-term outcomes after TBI where pre-injury characteristics and post-injury cognitive and physical characteristics account for the greatest proportion of explained variance. PMID- 16690585 TI - Effectiveness and expectations of intensive training: a comparison between child and youth rehabilitation and conductive education. AB - OBJECTIVE: To compare the effectiveness of two intensive training-programmes from a professional and parent perspective. To describe and compare the type of expectations of the two intensive training programmes with the self-reported individualized goals. DESIGN: Quasi-experimental with two groups. SETTING AND INTERVENTION: Traditional health care and conductive education. PATIENTS AND THEIR PARENTS: Fifty-four children with cerebral palsy, 3-16 years old. METHODS: Data included a self-reported individualized goal measure (SRIGM), before and after the ITP. Individualized goals were classified according to the International Classification of Functioning (ICF). Clinical measures (CM) included repeated measures with Gross Motor Function Measure (GMFM) and Pediatric Evaluation of Disability Inventory-Functional Skills (PEDI-FS). RESULTS: Twenty eight parents out of 54 perceived a clinically significant improvement on the SRIGM with no significant difference between the training programmes. Most individualized goals were formulated in the domain of Mobility (115 out of 248) and Neuromusculoskeletal and movement-related functions (64 out of 248 goals) of ICF in both training programmes. There was no difference in the proportion of improvement measured with SRIGM compared to the CM, if an improvement in any dimension in GMFM or domain in PEDI FS was counted. CONCLUSION: There were no major differences in outcome and expectations between the training programmes. Parents' expectations were mainly directed towards improvement in prerequisites of motor function and mobility skills. The SRIGM confirmed the outcome on the CM. PMID- 16690586 TI - Characterisation of unilateral neglect by physiotherapists. AB - PURPOSE: It is not known how clinicians characterise unilateral neglect (ULN) or whether they consider the different types of ULN during their evaluation and management of patients with this condition. The purpose of this study was to gain insight into physiotherapists' understanding of ULN and to identify the terminology used by clinicians to characterise neglect behaviour. METHOD: Qualitative research design employing focus groups and one-to-one interviews. Thirty-three experienced neurological and novice physiotherapists from one Australian state were asked to discuss how they characterise ULN and to explain what they meant by the terms they used. Data analysis involved preparation of verbatim transcripts followed by coding, data reduction, and identification of major themes. Data management was facilitated using NVivo computer software. FINDINGS: Terms such as sensory neglect, visual neglect, extinction, inattention, and functional neglect were used to characterise neglect behaviour but there was considerable confusion between many of these terms. Motor neglect was an unfamiliar concept to many participants. Functional implications of ULN were considered important. Characterisations of neglect according to the spatial distribution of the behaviour were uncommon. CONCLUSIONS: Despite awareness of many types of neglect, the physiotherapists in this study did not have a clear understanding of how to accurately characterise the different types of ULN. Education to address the issue is required. PMID- 16690587 TI - The construct validity of a spasticity measurement device for clinical practice: an alternative to the Ashworth scales. AB - INTRODUCTION: Spasticity is a significant cause of disability in people with an upper motor neurone lesion, but there is a paucity of appropriate outcome measures to evaluate this phenomenon. The aim was to test the construct validity of a clinically relevant, non-invasive measure of spasticity. METHODS: A cross section study design in which participants with elbow flexor spasticity and capable of providing written informed consent were recruited. RESULTS: Fourteen stroke patients participated (six female and eight male). Median age was 61 years and the median time post stroke was 48 months. Six patients had a MAS grading of '1+', three a grade of '2' and five a grade of '3'. The velocity of the brisk stretch was significantly higher than that of the slow stretch (p < 0.05: median difference, 34 degrees /s: IQR, 20 - 46). Flexor muscle activity during the brisk stretch was significantly higher than that of the slow stretch (p < 0.05: median difference, 2.0 microV; IQR, 0.4 - 8.4). In contrast the RPE was not significantly different between the slow and the fast stretches (p > 0.1: median difference, 0.07 N/deg; IQR, - 0.09 - 0.16). There were no patterns of association between the MAS, elbow flexor muscle activity and RPE. Other important observations, in some patients, were: continuous background muscle activation consistent with descriptions of spastic dystonia; muscle activity at the slow velocity stretch; muscle activation patterns consistent with the clasp knife phenomenon. CONCLUSIONS: The measurement system was capable of measuring spasticity as defined by Lance (1980; In: Lance et al., editors. Spasticity: disordered motor control. Chicago, IL: Year Book. p 185 - 204). In addition, it enabled various other clinical phenomena associated with spasticity to be measured. Assessing spasticity by measuring changes in resistance to passive movement only may not be sufficient, as the latter is influenced by many factors of which spasticity may only be one. Further work is now required to investigate repeatability and sensitivity. PMID- 16690588 TI - The application of complex adaptive systems theory to clinical practice in rehabilitation. AB - It is increasingly emphasised that effective healthcare for the growing number of chronic disease and lifestyle issues must be grounded in a non-reductionist paradigm focused on understanding relationships and applying flexible problem solving. To address these needs, key principles of complex adaptive systems theory (CAS) are being applied to healthcare planning and research. The aim of this clinical commentary is to provide a brief overview of complex adaptive systems theory and illustrate its relevance to rehabilitation professionals. Further, the review will explore occupational therapist and physiotherapist profession-specific publications for evidence of CAS theory applications that can be used to illustrate the clinical usefulness of complexity science concepts. The commentary will also identify other resources that can assist healthcare providers in developing an understanding of the framework's assumptions and principles for affecting change. Lastly, the implications of failing to acquire a competent understanding of CAS principles and command of the terminology will be addressed and therapists challenged to assume an equal role in mapping healthcare delivery in the 21st century. PMID- 16690589 TI - Potential contributions to disability theorizing and research from positive pyschology. AB - PURPOSE: Within the context of taking the perspectives of disabled individuals themselves more seriously, it is imperative that researchers and practitioners accord due primacy to the experiential worlds and everyday lived experiences of individuals with disabilities. Based on the premise that theoretical groundedness (along with methodological rigour) is integral to sound research, it is argued that the existing body of knowledge regarding the lived experiences of individuals with disabilities may be complemented by a conceptual placing of understandings of disability in relation to a prominent theoretical pathway at the moment, namely, the interdisciplinary emphasis on human strengths and wellness. METHOD: A critical engagement with key conceptualizations of disability entailed a focus on the development, strengths, and challenges of medical, social and psychologically based conceptualizations of disability. RESULTS AND CONCLUSION: The existing body of knowledge around disability may be extended and complemented by key concepts regarding human health/wellness from the field of Positive Psychology. Positive Psychology's keynote contribution to the study of disability and rehabilitation entails directing researchers and practitioners to the aim of building, reinforcing and extending disabled individuals' strengths and capacities in order to optimize their functioning in all areas of life, and thereby promote wellness. PMID- 16690590 TI - Low folate status increases chromosomal damage by X-ray irradiation. AB - PURPOSE: To examine how folate status influences chromosomal damage following X ray irradiation. MATERIAL AND METHODS: In an animal study, mice were fed either a low, basal, or high folic acid diet (0, 2, or 40 mg/kg diet, respectively) for 4 weeks, and then given total body irradiation (TBI) at 0.5 Gy. In a human study, subjects were supplemented with folic acid (800 microg/day) for 2 weeks and their peripheral blood was irradiated at 0.5 Gy in vitro. Chromosomal damage was determined by micronucleus assay. RESULTS: In an animal study, TBI-induced chromosomal damage was higher and folate concentration was lower in the bone marrow of the low folic acid group compared to the other two diet groups. The chromosomal damage and folate concentration were comparable between the basal and high folic acid groups. TBI administered to mice decreased folate in the plasma, erythrocyte and bone marrow. In a human study, supplementation with folic acid increased plasma folate, but did not influence either plasma homocysteine or X ray-induced chromosomal damage in lymphocytes. CONCLUSION: Low folate status increases susceptibility to X-ray-induced chromosomal damage, but excessive folic acid supplementation under normal conditions yields no further protection due to folate saturation in the target tissue. PMID- 16690591 TI - Lack of association between acute exposure to ionizing radiation and liver cirrhosis. AB - PURPOSE: Although previous studies have shown significantly increased risks of liver cirrhosis and chronic liver disease for acute radiation exposure among survivors of the atomic bombings of Hiroshima and Nagasaki, Japan, these studies have not taken into account hepatitis B virus (HBV) infections. Because HBV is associated with both A-bomb radiation and liver cirrhosis, our goal was to investigate the relationship of acute ionizing radiation to liver cirrhosis adjusting for HBV, co-occurring primary liver cancer (PLC), and other potential confounders. MATERIALS AND METHODS: Using a cross-sectional design and pathology review of a cohort of Japanese atomic-bomb survivors, we found that 213 of 335 (63.6%) subjects with PLC and 55 of 776 (7.1%) subjects without PLC had cirrhosis. RESULTS: We found no association between acute exposure to A-bomb radiation and liver cirrhosis. The adjusted odds ratio of cirrhosis per Sv liver irradiation was 0.59 (95% confidence interval: 0.27 - 1.27). Cirrhosis risks for the highest tertile of radiation exposure (mean exposure 0.7 Sv) were also not elevated (0.8, 0.26 - 2.12 and 0.2, 0.03 - 0.98 among subjects with and without PLC. CONCLUSIONS: Acute exposure to liver irradiation does not increase risks of liver cirrhosis, regardless of PLC status. PMID- 16690592 TI - Cloning and characterization of an ascidian homolog of the human 8-oxoguanine DNA glycosylase (Ogg1) that is involved in the repair of 8-oxo-7,8-dihydroguanine in DNA in Ciona intestinalis. AB - PURPOSE: It is of interest to perform a systematic comparative analysis of the conserved domains in DNA glycosylases and the evolution of DNA base excision repair systems. Furthermore, it is important to characterize the roles and regulation of base excision repair during the development of organisms. To address these issues, we first identified 8-oxo-7,8-dihydroguanine (8-oxoG)-DNA glycosylase (Ogg1) of the ascidian Ciona intestinalis as a good model system. MATERIALS AND METHODS: A cDNA clone coding for a peptide with homology to human Ogg1 was identified in the expressed sequence tag (EST) database from the Ciona cDNA resources. We examined whether CiOgg1 has DNA glycosylase/AP (apurinic/apyrimidinic) lyase activities for 8-oxoG-containing oligonucleotide. Furthermore, the expression level of CiOgg1 was compared in various tissues of Ciona intestinalis. RESULTS: The CiOgg1gene encoded a protein of 351 amino acids, which shows 37% identity of amino acid sequence with human Ogg1. The Helix hairpin-Helix motif was highly conserved. The ascidian enzyme had functional 8 oxoG-DNA glycosylase/AP lyase activities, which removed 8-oxoG opposite cytosine from DNA. Expression of the CiOgg1 significantly reduced the frequency of spontaneous G:C to T:A transversions in E. coli mutM mutY. The highest expression level was observed in testis in Ciona intestinalis. CONCLUSIONS: The structure and functions of Ogg1 are well conserved in Ciona intestinalis. CiOgg1 is involved in the repair of 8-oxoG in DNA in Ciona intestinalis. PMID- 16690593 TI - Radiobiological analysis of human melanoma cells on the 62 MeV CATANA proton beam. AB - PURPOSE: To measure the ability of protons and gamma-rays to effect cell viability and cell survival of human HTB140 melanoma cells. MATERIALS AND METHODS: Exponentially growing HTB140 cells were irradiated close to the Bragg peak maximum of the 62 MeV protons or with 60Co gamma-rays with single doses, ranging from 8 - 24 Gy. Cell viability using the 3-(4,5-dimethylthiazol-2yl)-2,5 diphenyltetrazolium bromide (MTT) assay was evaluated at 6 h, 24 h, 48 h or 7 days after irradiation and clonogenic survival was assessed at 7 days after irradiation. Cell cycle phase redistribution and the level of apoptosis were evaluated at 6 h and 48 h after irradiation. RESULTS: The study of cell viability as a function of time (cell survival progression) and cell survival, using a clonal assay, demonstrated the considerably stronger inactivation effect of protons compared to gamma-rays with a relative biological effectiveness (RBE) of approximately 1.64. Cell cycle phase distribution and apoptosis levels with time enabled us to investigate the development and the character of the damage induced by irradiation. Due to the high radio-resistance of HTB140 cells, cell cycle phase redistribution exhibited only a modest cell accumulation in G2/M phase. Protons but not gamma-rays induced apoptosis. CONCLUSIONS: It appears that protons reduce the number of HTB140 cells by apoptosis as well as by severe DNA damage, while gamma-rays eliminate viable cells primarily by the production of irreparable DNA damage. Protons have an increased RBE relative to gamma-rays. PMID- 16690594 TI - Microwave exposure of neuronal cells in vitro: Study of apoptosis. AB - PURPOSE: The aim of this study was to investigate microwave (MW) effects on neuronal apoptosis in vitro. MATERIALS AND METHODS: Human neuroblastoma cells SH SY5Y were exposed to a 900 MHz global system for mobile communication (GSM) or continuous-wave (CW) radiofrequency fields for 24 h in a wire-patch cell. The specific absorption rates (SAR) used were 2 W/kg for CW and 0.25 W/kg average for GSM. During CW exposure, an increase of 2 degrees C was measured, and controls with cells exposed to 39 degrees C were then performed. Apoptosis rate was assessed immediately or 24 h after exposure using three methods: (i) 4',6-diamino 2-phenylindole (DAPI) staining; (ii) flow cytometry using double staining with TdT-mediated dUTP nick-end labeling (TUNEL) and propidium iodide (PI); and (iii) measurement of caspase-3 activity by fluorimetry. RESULTS: No statistically significant difference in the apoptosis rate was observed between sham and 24 h MW-exposed cells, either GSM-900 at an average SAR of 0.25 W/kg, or CW 900 MHz at a SAR of 2 W/kg, either 0 h or 24 h post-exposure. Furthermore, for CW-exposure, apoptosis rates were comparable between sham-, CW-, 37 degrees C- and 39 degrees C-exposed cells. All three methods used to assess apoptosis were concordant. CONCLUSION: These results showed that, under the conditions of the present experiment, MW-exposure (either CW or GSM-900) does not significantly increase the apoptosis rate in the human neuroblastoma cell line SH-SY5Y. PMID- 16690595 TI - Radiosensitization of tumor cells by modulation of ATM kinase. AB - PURPOSE: To elucidate the relationship between the radiation-induced activation of ataxia telangiectasia mutated (ATM) kinase, G2 arrest and the caffeine-induced radiosensitization. METHOD: RKO cells (human colorectal cancer cells) and ATM kinase over-expressing RKO/ATM cells were used. The cellular radiosensitivity was determined with clonogenic survival assay and the cell cycle progression, including G2 arrest, was studied with flow cytometry. The activity of ATM kinase, check point 2 (Chk2) kinase and cycline B1/cell division cycle 2 (Cdc2) kinase was investigated. The radiosensitivity of RKO xenografts grown in nude mice was studied. RESULTS: RKO/ATM cells were radioresistant as compared with RKO cells. There was a greater increase in ATM kinase activity and G2 arrest in RKO/ATM cells than in RKO cells. Caffeine also sensitized both RKO cells and RKO/ATM cells to radiation. The caffeine treatment suppressed the radiation-induced activation of ATM kinase, suppressed the activation of Chk2 kinase and inhibited the accumulation of cells in G2 phase. The activity of cycline B1/Cdc2 kinase increased earlier but decayed rapidly in the presence of caffeine. Caffeine enhanced radiation-induced growth delay of RKO xenografts. CONCLUSIONS: Caffeine inhibited the radiation-induced activation of ATM kinase, thereby preventing the accumulation of cells in G2 phase. Consequently, radiosensitivity of cells increased in the presence of caffeine both in vitro and in vivo. PMID- 16690596 TI - Long-term exposure of Sprague Dawley rats to 20 kHz triangular magnetic fields. AB - PURPOSE: There are only a few reports on harmful effects of 20 kHz sine waves; however, it is essential to comprehensively evaluate the potentially harmful effect of triangular signals at the same frequency. Therefore, in this study, effects of long-term exposure to 20 kHz magnetic fields was examined. MATERIALS AND METHODS: Eighty Sprague Dawley rats were divided into two groups (half male and female in each sham and exposed groups), and they were exposed to 20 kHz triangular magnetic fields at 6.25 microT rms for 8 h/day for 12 or 18 months. Urinalysis [pH, glucose, protein, ketone bodies, red blood cells (RBC), nitrogen, bilirubin, urobilinogen, and specific gravity], hematological analysis (RBC, hemoglobin, hematocrit, thrombocyte count, and leucocyte count), blood biochemistry (total protein, blood urea nitrogen, creatinine, glucose, total bilirubin, total cholesterol, aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase), and histopathological analysis of organs (thymus, stomach, intestine, liver, kidney, testis, ovary, spleen, brain, heart, and lung) were performed. RESULTS: No significant differences were seen between 20 kHz magnetic-fields exposed rats and sham-exposed rats in body and organ weights, hematological analysis, blood biochemistry, urinalysis data, and histopathological examination, except for the numbers of neutrophiles and lymphocytes in female rats. The number of neutrophiles was significantly increased in female rats on the 12th month after exposure, and the number of lymphocytes in female rats was significantly decreased on the 18th month. CONCLUSION: Long-term exposure of rats to 20 kHz triangular magnetic fields did not induce any significantly harmful effects, except changes in neutrophiles at 12 months and lymphocytes at 18 months of exposure in female rats. These hematological changes need to be investigated again at a higher intensity of 20 kHz magnetic fields. PMID- 16690597 TI - Chromatin loops are responsible for higher counts of small DNA fragments induced by high-LET radiation, while chromosomal domains do not affect the fragment sizes. AB - PURPOSE: To apply a polymer model of DNA damage induced by high-LET (linear energy transfer) radiation and determine the influence of chromosomal domains and loops on fragment length distribution. MATERIALS AND METHODS: The yields of DSB (double-strand breaks) induced by high-LET radiation were calculated using a track structure model along with a polymer model of DNA packed in the cell nucleus. The cell nucleus was constructed to include the chromosomal domains and chromatin loops. The latter were generated by the random walk method. RESULTS AND CONCLUSIONS: We present data for DSB yields per track per cell, DNA fragment sizes, the radial distribution of DSB with respect to the track center, and the distribution of 0, 1, 2, and more DSB from a single particle. Calculations were carried out for a range of particles including He (40 keV/microm), N (225 keV/microm), and Fe ions (150 keV/mum). Situations relevant to PFGE (pulsed-field gel electrophoresis) and microbeam experiments with direct irradiation of the cell nucleus were simulated to demonstrate the applicability of the model. Data show that chromosomal domains do not have a significant influence on fragment size distribution, while the presence of DNA loops increases the frequencies of smaller fragments by nearly 30% for fragment sizes in the range from 2 kbp (bp = base pair) to 20 kbp. PMID- 16690604 TI - MutL-catalyzed ATP hydrolysis is required at a post-UvrD loading step in methyl directed mismatch repair. AB - Methyl-directed mismatch repair is a coordinated process that ensures replication fidelity and genome integrity by resolving base pair mismatches and insertion/deletion loops. This post-replicative event involves the activities of several proteins, many of which appear to be regulated by MutL. MutL interacts with and modulates the activities of MutS, MutH, UvrD, and perhaps other proteins. The purified protein catalyzes a slow ATP hydrolysis reaction that is essential for its role in mismatch repair. However, the role of the ATP hydrolysis reaction is not understood. We have begun to address this issue using two point mutants: MutL-E29A, which binds nucleotide but does not catalyze ATP hydrolysis, and MutL-D58A, which does not bind nucleotide. As expected, both mutants failed to complement the loss of MutL in genetic assays. Purified MutL E29A protein interacted with MutS and stimulated the MutH-catalyzed nicking reaction in a mismatch-dependent manner. Importantly, MutL-E29A stimulated the loading of UvrD on model substrates. In fact, stimulation of UvrD-catalyzed unwinding was more robust with MutL-E29A than the wild-type protein. MutL-D58A, on the other hand, did not interact with MutS, stimulate MutH-catalyzed nicking, or stimulate the loading of UvrD. We conclude that ATP-bound MutL is required for the incision steps associated with mismatch repair and that ATP hydrolysis by MutL is required for a step in the mismatch repair pathway subsequent to the loading of UvrD and may serve to regulate helicase loading. PMID- 16690605 TI - Effects of human deafness gamma-actin mutations (DFNA20/26) on actin function. AB - Six point mutations in non-muscle gamma-actin at the DFNA20/26 locus cause autosomal dominant nonsyndromic hearing loss. The molecular basis for the hearing loss is unknown. We have engineered each gamma-actin mutation into yeast actin to investigate the effects of these mutations on actin function in vivo and in vitro. Cells expressing each of the mutant actins as the sole actin in the cell were viable. Four of the six mutant strains exhibited significant growth deficiencies in complete medium and an inability to grow on glycerol as the sole carbon source, implying a mitochondrial defect(s). These four strains exhibited abnormal mitochondrial morphology, although the mtDNA was retained. All of the mutant cells exhibited an abnormally high percentage of fragmented/non-polarized actin cables or randomly distributed actin patches. Five of the six mutants displayed strain-specific vacuole morphological abnormalities. Two of the purified mutant actins exhibited decreased thermal stability and increased rates of nucleotide exchange, indicative of increased protein flexibility. V370A actin alone polymerized abnormally. It aggregated in low ionic strength buffer and polymerized faster than wild-type actin, probably in part because of enhanced nucleation. Mixtures of wild-type and V370A actins displayed kinetic properties in proportion to the mole fraction of each actin in the mixture. No dominant effect of the mutant actin was observed. Our results suggest that a major factor in the deafness caused by these mutations is an altered ability of the actin filaments to be properly regulated by actin-binding proteins rather than an inability to polymerize. PMID- 16690606 TI - Transforming growth factor-beta stimulates cyclin D1 expression through activation of beta-catenin signaling in chondrocytes. AB - Transforming growth factor-beta (TGF-beta) plays an essential role in chondrocyte maturation. It stimulates chondrocyte proliferation but inhibits chondrocyte differentiation. In this study, we found that TGF-beta rapidly induced beta catenin protein levels and signaling in murine neonatal sternal primary chondrocytes. TGF-beta-increased beta-catenin induction was reproduced by overexpression of SMAD3 and was absent in Smad3(-/-) chondrocytes treated with TGF-beta. SMAD3 inhibited beta-transducin repeat-containing protein-mediated degradation of beta-catenin and immunoprecipitated with beta-catenin following TGF-beta treatment. Both SMAD3 and beta-catenin co-localized to the nucleus after TGF-beta treatment. Although both TGF-beta and beta-catenin stimulated cyclin D(1) expression in chondrocytes, the effect of TGF-beta was inhibited with beta catenin gene deletion or SMAD3 loss of function. These results demonstrate that TGF-beta stimulates cyclin D(1) expression at least in part through activation of beta-catenin signaling. PMID- 16690607 TI - Region 3.2 of the sigma subunit contributes to the binding of the 3'-initiating nucleotide in the RNA polymerase active center and facilitates promoter clearance during initiation. AB - Region 3.2 of the RNA polymerase sigma subunit forms a loop that protrudes toward RNA polymerase active center and partially blocks RNA exit channel. To provide some insights into the functional role of this region, we studied a deletion variant of the Escherichia coli sigma(70) subunit that lacked amino acids 513-519 corresponding to the tip of the loop. The deletion had multiple effects on transcription initiation including: (i) a significant decrease in the amount of short abortive RNAs synthesized during initiation, (ii) defects in promoter escape, (iii) loss of the contacts between the sigma subunit and the nascent RNA during initiation and, finally, (iv) dramatic increase in the K(m) value for the 3'-initiating nucleotide. At the same time, the mutation did not impair promoter opening and the binding of the 5'-initiating purine nucleotide. In summary, our data demonstrate an important role of sigma region 3.2 in the binding of initiating substrates in RNA polymerase active center and in the process of promoter clearance. PMID- 16690608 TI - Overexpression of a cytochrome b5 reductase-like protein causes kinetoplast DNA loss in Trypanosoma brucei. AB - The mitochondrial genome of trypanosomes, termed kinetoplast DNA (kDNA), contains thousands of minicircles and dozens of maxicircles topologically interlocked in a network. To identify proteins involved in network replication, we screened an inducible RNA interference-based genomic library for cells that lose kinetoplast DNA. In one cloned cell line with inducible kinetoplast DNA loss, we found that the RNA interference vector had aberrantly integrated into the genome resulting in overexpression of genes down-stream of the integration site (Motyka, S. A., Zhao, Z., Gull, K., and Englund, P. T. (2004) Mol. Biochem. Parasitol. 134, 163 167). We now report that the relevant overexpressed gene encodes a mitochondrial cytochrome b(5) reductase-like protein. This overexpression caused kDNA loss by oxidation/inactivation of the universal minicircle sequence-binding protein, which normally binds the minicircle replication origin and triggers replication. The rapid loss of maxicircles suggests that the universal minicircle sequence binding protein might also control maxicircle replication. Several lines of evidence indicate that the cytochrome b(5) reductase-like protein controls the oxidization status of the universal minicircle sequence-binding protein via tryparedoxin, a mitochondrial redox protein. For example, overexpression of mitochondrial tryparedoxin peroxidase, which utilizes tryparedoxin, also caused oxidation of the universal minicircle sequence-binding protein and kDNA loss. Furthermore, the growth defect caused by overexpression of cytochrome b(5) reductase-like protein could be partially rescued by simultaneously overexpressing tryparedoxin. PMID- 16690609 TI - Inhibition of transforming growth factor beta-enhanced serum response factor dependent transcription by SMAD7. AB - Transforming growth factor (TGF)-beta is present in large amounts in the airways of patients with asthma and with other diseases of the lung. We show here that TGFbeta treatment increased transcriptional activation of SM22alpha, a smooth muscle-specific promoter, in airway smooth muscle cells, and we demonstrate that this effect stems in part from TGFbeta-induced enhancement of serum response factor (SRF) DNA binding and transcription promoting activity. Overexpression of Smad7 inhibited TGFbeta-induced stimulation of SRF-dependent promoter function, and chromatin immunoprecipitation as well as co-immunoprecipitation assays established that endogenous or recombinant SRF interacts with Smad7 within the nucleus. The SRF binding domain of Smad7 mapped to the C-terminal half of the Smad7 molecule. TGFbeta treatment weakened Smad7 association with SRF, and conversely the Smad7-SRF interaction was increased by inhibition of the TGFbeta pathway through overexpression of a dominant negative mutant of TGFbeta receptor I or of Smad3 phosphorylation-deficient mutant. Our findings thus reveal that SRF Smad7 interactions in part mediate TGFbeta regulation of gene transcription in airway smooth muscle. This offers potential targets for interventions in treating lung inflammation and asthma. PMID- 16690610 TI - Polyamine depletion increases cytoplasmic levels of RNA-binding protein HuR leading to stabilization of nucleophosmin and p53 mRNAs. AB - Polyamines are essential for maintaining normal intestinal epithelial integrity, an effect that relies, at least in part, on their ability to keep low levels of nucleophosmin (NPM) and p53 mRNAs. The RNA-binding protein HuR associates with the p53 mRNA, as reported previously, and with the NPM mRNA, computationally predicted to be a target of HuR. Here, we show that HuR binds the NPM and p53 3' untranslated regions and stabilizes these mRNAs in polyamine-depleted intestinal epithelial cells. Depletion of cellular polyamines by inhibiting ornithine decarboxylase with alpha-difluoromethylornithine dramatically enhanced the cytoplasmic abundance of HuR, whereas ectopic ornithine decarboxylase overexpression decreased cytoplasmic HuR; neither intervention changed whole-cell HuR levels. HuR was found to specifically bind the 3'-untranslated regions of NPN and p53 mRNAs. HuR silencing rendered the NPM and p53 mRNAs unstable and prevented increases in NPM and p53 mRNA and protein in polyamine-deficient cells. These results indicate that polyamines modulate cytoplasmic HuR levels in intestinal epithelial cells, in turn controlling the stability of the NPM and p53 mRNAs and influencing NPM and p53 protein levels. PMID- 16690611 TI - Molecular mechanism underlying inverse agonist of angiotensin II type 1 receptor. AB - To delineate the molecular mechanism underlying the inverse agonist activity of olmesartan, a potent angiotensin II type 1 (AT1) receptor antagonist, we performed binding affinity studies and an inositol phosphate production assay. Binding affinity of olmesartan and its related compounds to wild-type and mutant AT1 receptors demonstrated that interactions between olmesartan and Tyr113, Lys199, His256, and Gln257 in the AT1 receptor were important. The inositol phosphate production assay of olmesartan and related compounds using mutant receptors indicated that the inverse agonist activity required two interactions, that between the hydroxyl group of olmesartan and Tyr113 in the receptor and that between the carboxyl group of olmesartan and Lys199 and His256 in the receptor. Gln257 was found to be important for the interaction with olmesartan but not for the inverse agonist activity. Based on these results, we constructed a model for the interaction between olmesartan and the AT1 receptor. Although the activation of G protein-coupled receptors is initiated by anti-clockwise rotation of transmembrane (TM) III and TM VI followed by changes in the conformation of the receptor, in this model, cooperative interactions between the hydroxyl group and Tyr113 in TM III and between the carboxyl group and His256 in TM VI were essential for the potent inverse agonist activity of olmesartan. We speculate that the specific interaction of olmesartan with these two TMs is essential for stabilizing the AT1 receptor in an inactive conformation. A better understanding of the molecular mechanisms of the inverse agonism could be useful for the development of new G protein-coupled receptor antagonists with inverse agonist activity. PMID- 16690612 TI - EWI-2 and EWI-F link the tetraspanin web to the actin cytoskeleton through their direct association with ezrin-radixin-moesin proteins. AB - EWI-2 and EWI-F, two members of a novel subfamily of Ig proteins, are direct partners of tetraspanins CD9 (Tspan29) and CD81 (Tspan28). These EWI proteins contain a stretch of basic charged amino acids in their cytoplasmic domains that may act as binding sites for actin-linking ezrin-radixin-moesin (ERM) proteins. Confocal microscopy analysis revealed that EWI-2 and EWI-F colocalized with ERM proteins at microspikes and microvilli of adherent cells and at the cellular uropod in polarized migrating leukocytes. Immunoprecipitation studies showed the association of EWI-2 and EWI-F with ERM proteins in vivo. Moreover, pulldown experiments and protein-protein binding assays with glutathione S-transferase fusion proteins containing the cytoplasmic domains of EWI proteins corroborated the strong and direct interaction between ERMs and these proteins. The active role of ERMs was further confirmed by double transfections with the N-terminal domain of moesin, which acts as a dominant negative form of ERMs, and was able to delocalize EWIs from the uropod of polarized leukocytes. In addition, direct association of EWI partner CD81 C-terminal domain with ERMs was also demonstrated. Functionally, silencing of endogenous EWI-2 expression by short interfering RNA in lymphoid CEM cells augmented cell migration, cellular polarity, and increased phosphorylation of ERMs. Hence, EWI proteins, through their direct interaction with ERM proteins, act as linkers to connect tetraspanin associated microdomains to actin cytoskeleton regulating cell motility and polarity. PMID- 16690613 TI - The epithelial Na+ channel is inhibited by a peptide derived from proteolytic processing of its alpha subunit. AB - Epithelial sodium channels (ENaCs) mediate Na(+) entry across the apical membrane of high resistance epithelia that line the distal nephron, airway and alveoli, and distal colon. These channels are composed of three homologous subunits, termed alpha, beta, and gamma, which have intracellular amino and carboxyl termini and two membrane-spanning domains connected by large extracellular loops. Maturation of ENaC subunits involves furin-dependent cleavage of the extracellular loops at two sites within the alpha subunit and at a single site within the gamma subunit. The alpha subunits must be cleaved twice, immediately following Arg-205 and Arg-231, in order for channels to be fully active. Channels lacking alpha subunit cleavage are inactive with a very low open probability. In contrast, channels lacking both alpha subunit cleavage and the tract alphaAsp-206 Arg-231 are active when expressed in oocytes, suggesting that alphaAsp-206-Arg 231 functions as an inhibitor that stabilizes the channel in the closed conformation. A synthetic 26-mer peptide (alpha-26), corresponding to alphaAsp 206-Arg-231, reversibly inhibits wild-type mouse ENaCs expressed in Xenopus oocytes, as well as endogenous Na(+) channels expressed in either a mouse collecting duct cell line or primary cultures of human airway epithelial cells. The IC(50) for amiloride block of ENaC was not affected by the presence of alpha 26, indicating that alpha-26 does not bind to or interact with the amiloride binding site. Substitution of Arg residues within alpha-26 with Glu, or substitution of Pro residues with Ala, significantly reduced the efficacy of alpha-26. The peptide inhibits ENaC by reducing channel open probability. Our results suggest that proteolysis of the alpha subunit activates ENaC by disassociating an inhibitory domain (alphaAsp-206-Arg-231) from its effector site within the channel complex. PMID- 16690614 TI - Characterization of the tandem GAF domain of human phosphodiesterase 5 using a cyanobacterial adenylyl cyclase as a reporter enzyme. AB - We analyzed cGMP signaling by the human phosphodiesterase 5 (hPDE5) tandem GAF domain based on a functional activation assay. The C-terminal catalytic domain of the cyanobacterial adenylyl cyclase (AC) cyaB1 was used as a reporter enzyme. We demonstrate functional coupling between the hPDE5 GAF ensemble and the AC resulting in a chimera stimulated 10-fold by cGMP. The hPDE5 GAF domain has an inhibitory effect on AC activity, which is released upon cGMP activation. Removal of 109 amino acids from the N terminus resulted in partial disengagement of the GAF domain and AC, i.e. in a 10-fold increase in basal activity, and affected cGMP affinity. The Ser-102 phosphorylation site of hPDE5 increased cGMP affinity, as shown by a 5-fold lower K(D) for cGMP in a S102D mutant, which mimicked complete modification. The function of the NKFDE motif, which is a signature of all GAF domains with known cyclic nucleotide binding capacity, was elucidated by targeted mutations. Data with either single and double mutants in either GAF A or GAF B or a quadruple mutant affecting both subdomains simultaneously indicated that it is impossible to functionally assign cGMP binding and intramolecular signaling to either GAF A or B of hPDE5. Both subdomains are structurally and functionally interdependent and act in concert in regulating cycaB1 AC and, most likely, also hPDE5. PMID- 16690615 TI - The mucin Muc4 potentiates neuregulin signaling by increasing the cell-surface populations of ErbB2 and ErbB3. AB - Mucins provide a protective barrier for epithelial surfaces, and their overexpression in tumors has been implicated in malignancy. We have previously demonstrated that Muc4, a transmembrane mucin that promotes tumor growth and metastasis, physically interacts with the ErbB2 receptor tyrosine kinase and augments receptor tyrosine phosphorylation in response to the neuregulin-1beta (NRG1beta) growth factor. In the present study we demonstrate that Muc4 expression in A375 human melanoma cells, as well as MCF7 and T47D human breast cancer cells, enhances NRG1beta signaling through the phosphatidylinositol 3 kinase pathway. In examining the mechanism underlying Muc4-potentiated ErbB2 signaling, we found that Muc4 expression markedly augments NRG1beta binding to A375 cells without altering the total quantity of receptors expressed by the cells. Cell-surface protein biotinylation experiments and immunofluorescence studies suggest that Muc4 induces the relocalization of the ErbB2 and ErbB3 receptors from intracellular compartments to the plasma membrane. Moreover, Muc4 interferes with the accumulation of surface receptors within internal compartments following NRG1beta treatment by suppressing the efficiency of receptor internalization. These observations suggest that transmembrane mucins can modulate receptor tyrosine kinase signaling by influencing receptor localization and trafficking and contribute to our understanding of the mechanisms by which mucins contribute to tumor growth and progression. PMID- 16690616 TI - Novel FXXFF and FXXMF motifs in androgen receptor cofactors mediate high affinity and specific interactions with the ligand-binding domain. AB - Upon hormone binding, a hydrophobic coactivator binding groove is induced in the androgen receptor (AR) ligand-binding domain (LBD). This groove serves as high affinity docking site for alpha-helical FXXLF motifs present in the AR N-terminal domain and in AR cofactors. Study of the amino acid requirements at position +4 of the AR FXXLF motif revealed that most amino acid substitutions strongly reduced or completely abrogated AR LBD interaction. Strong interactions were still observed following substitution of Leu+4 by Phe or Met residues. Leu+4 to Met or Phe substitutions in the FXXLF motifs of AR cofactors ARA54 and ARA70 were also compatible with strong AR LBD binding. Like the corresponding FXXLF motifs, interactions of FXXFF and FXXMF variants of AR and ARA54 motifs were AR specific, whereas variants of the less AR-selective ARA70 motif displayed increased AR specificity. A survey of currently known AR-binding proteins revealed the presence of an FXXFF motif in gelsolin and an FXXMF motif in PAK6. In vivo fluorescence resonance energy transfer and functional protein-protein interaction assays showed direct, efficient, and specific interactions of both motifs with AR LBD. Mutation of these motifs abrogated interaction of gelsolin and PAK6 proteins with AR. In conclusion, we have demonstrated strong interaction of FXXFF and FXXMF motifs to the AR coactivator binding groove, thereby mediating specific binding of a subgroup of cofactors to the AR LBD. PMID- 16690617 TI - Regulation of neuronal nitric-oxide synthase activity by somatostatin analogs following SST5 somatostatin receptor activation. AB - Somatostatin receptor SST5 is an inhibitory G protein-coupled receptor that exerts a strong cytostatic effect on various cell types. We reported previously that the SST5 anti-proliferative effect results in the inhibition of mitogen induced increases in intracellular cGMP levels and MAPK activity. This study was conducted to define the early molecular events accountable for the SST5-mediated anti-proliferative effect. Here, we demonstrate that, in Chinese hamster ovary cells expressing SST5 (CHO/SST5 cells), somatostatin inhibited cell proliferation induced by nitric oxide donors and overexpression of the neuronal nitric-oxide synthase (nNOS) protein isoform. Accordingly, nNOS activity and dimerization were strongly inhibited following SST5 activation by the somatostatin analog RC-160. In CHO/SST5 cells, nNOS was dynamically recruited by the SST5 receptor and phosphorylated at tyrosyl residues following RC-160 treatment. RC-160 induced SST5-p60(src) kinase complex formation and subsequent p60(src) kinase activation. Coexpression of an inactive p60(src) kinase mutant with SST5 blocked RC-160 induced nNOS phosphorylation and inactivation and prevented the SST5-mediated anti-proliferative effect. In CHO/SST5 cells, p60(src) kinase associated with nNOS to induce its inactivation by phosphorylation at tyrosyl residues following RC-160 treatment. Using recombinant proteins, we demonstrated that such phosphorylation prevented nNOS homodimerization. Next, surface plasmon resonance and mutation analysis revealed that p60(src) directly associated with nNOS phosphorylated Tyr604. SST5-mediated inhibition of nNOS activity was demonstrated to be essential to the RC-160 anti-proliferative effect on pancreatic endocrine tumor-derived cells. We therefore identified nNOS as a new p60(src) kinase substrate essential for SST5-mediated anti-proliferative action. PMID- 16690618 TI - Structural changes of Escherichia coli ferric uptake regulator during metal dependent dimerization and activation explored by NMR and X-ray crystallography. AB - Ferric uptake regulator (Fur) is a global bacterial regulator that uses iron as a cofactor to bind to specific DNA sequences. Escherichia coli Fur is usually isolated as a homodimer with two metal sites per subunit. Metal binding to the iron site induces protein activation; however the exact role of the structural zinc site is still unknown. Structural studies of three different forms of the Escherichia coli Fur protein (nonactivated dimer, monomer, and truncated Fur-(1 82)) were performed. Dimerization of the oxidized monomer was followed by NMR in the presence of a reductant (dithiothreitol) and Zn(II). Reduction of the disulfide bridges causes only local structure variations, whereas zinc addition to reduced Fur induces protein dimerization. This demonstrates for the first time the essential role of zinc in the stabilization of the quaternary structure. The secondary structures of the mono- and dimeric forms are almost conserved in the N terminal DNA-binding domain, except for the first helix, which is not present in the nonactivated dimer. In contrast, the C-terminal dimerization domain is well structured in the dimer but appears flexible in the monomer. This is also confirmed by heteronuclear Overhauser effect data. The crystal structure at 1.8A resolution of a truncated protein (Fur-(1-82)) is described and found to be identical to the N-terminal domain in the monomeric and in the metal-activated state. Altogether, these data allow us to propose an activation mechanism for E. coli Fur involving the folding/unfolding of the N-terminal helix. PMID- 16690619 TI - Molecular rearrangements of the Kv2.1 potassium channel termini associated with voltage gating. AB - The voltage-gated Kv2.1 channel is composed of four identical subunits folded around the central pore and does not inactivate appreciably during short depolarizing pulses. To study voltage-induced relative molecular rearrangements of the channel, Kv2.1 subunits were genetically fused with enhanced cyan fluorescent protein and/or enhanced yellow fluorescent protein, expressed in COS1 cells, and investigated using fluorescence resonance energy transfer (FRET) microscopy combined with patch clamp. Fusion of fluorophores to either or both termini of the Kv2.1 monomer did not significantly affect the gating properties of the channel. FRET between the N- and C-terminal tags fused to the same or different Kv2.1 monomers decreased upon activation of the channel by depolarization from -80 to +60 mV, suggesting voltage-gated relative rearrangement between the termini. Because FRET between the Kv2.1 N- or C terminal tags and the membrane-trapped EYFP(N)-PH pleckstrin homology domains did not change on depolarization, voltage-gated relative movements between the Kv2.1 termini occurred in a plane parallel to the plasma membrane, within a distance of 1-10 nm. FRET between the N-terminal tags did not change upon depolarization, indicating that the N termini do not rearrange relative to each other, but they could either move cooperatively with the Kv2.1 tetramer or not move at all. No FRET was detected between the C-terminal tags. Assuming their randomized orientation in the symmetrically arranged Kv2.1 subunits, C termini may move outwards in order to produce relative rearrangements between N and C termini upon depolarization. PMID- 16690621 TI - Role of protein-tyrosine phosphatase SHP2 in focal adhesion kinase down regulation during neutrophil cathepsin G-induced cardiomyocytes anoikis. AB - Inflammatory cells and their proteases contribute to tissue reparation at site of inflammation. Although beneficial at early stages, excessive inflammatory reaction leads to cell death and tissue damage. Cathepsin G (Cat.G), a neutrophil derived serine protease, has been shown to induce neonatal rat cardiomyocyte detachment and apoptosis by anoikis through caspase-3 dependent pathway. However the early mechanisms that trigger Cat.G-induced caspase-3 activation are not known. This study identifies focal adhesion kinase (FAK) tyrosine dephosphorylation as an early mechanism that regulates Cat.G-induced anoikis in cardiomyocytes. Both FAK tyrosine phosphorylation at Tyr-397 and kinase activity decrease rapidly upon Cat.G treatment and was associated with a decrease of FAK association with adapter and cytoskeletal proteins, p130(Cas) and paxillin, respectively. FAK-decreased tyrosine phosphorylation is required for Cat.G induced myocyte anoikis as concurrent expression of phosphorylation-deficient FAK mutated at Tyr-397 or pretreatment with a protein-tyrosine phosphatase (PTP) inhibitor, pervanadate, blocks Cat.G-induced FAK tyrosine dephosphorylation, caspase-3 activation and DNA fragmentation. Analysis of PTPs activation shows that Cat.G treatment induces an increase of SHP2 and PTEN phosphorylation; however, only SHP2 forms a complex with FAK in response to Cat.G. Expression of dominant negative SHP2 mutant markedly attenuates FAK tyrosine dephosphorylation induced by Cat.G and protects myocytes to undergo apoptosis. In contrast, increased SHP2 expression exacerbates Cat.G-induced FAK tyrosine dephosphorylation and myocyte apoptosis. Taken together, these results show that Cat.G induces SHP2 activation that leads to FAK tyrosine dephosphorylation and promotes cardiomyocyte anoikis. PMID- 16690622 TI - Cj1121c, a novel UDP-4-keto-6-deoxy-GlcNAc C-4 aminotransferase essential for protein glycosylation and virulence in Campylobacter jejuni. AB - Campylobacter jejuni produces glycoproteins that are essential for virulence. These glycoproteins carry diacetamidobacillosamine (DAB), a sugar that is not found in humans. Hence, the enzymes responsible for DAB synthesis represent potential therapeutic targets. We describe the biochemical characterization of Cj1121c, a putative aminotransferase encoded by the general protein glycosylation locus, to assess its role in DAB biosynthesis. By using overexpressed and affinity-purified enzyme, we demonstrate that Cj1121c has pyridoxal phosphate- and glutamate-dependent UDP-4-keto-6-deoxy-GlcNAc C-4 transaminase activity and produces UDP-4-amino-4,6-dideoxy-GlcNAc. This is consistent with a role in DAB biosynthesis and distinguishes Cj1121c from Cj1294, a homologous UDP-2-acetamido 2,6-dideoxy-beta-l-arabino-4-hexulose C-4 aminotransferase that we characterized previously. We show that Cj1121c can also use this 4-keto-arabino sugar indirectly as a substrate, that Cj1121c and Cj1294 are active simultaneously in C. jejuni, and that the activity of Cj1121c is preponderant under standard growth conditions. Kinetic data indicate that Cj1121c has a slightly higher catalytic efficiency than Cj1294 with regard to the 4-keto-arabino substrate. By site directed mutagenesis, we show that residues Glu-158 and Leu-131 are not essential for catalysis or for substrate specificity contrary to expectations. We further demonstrate that a cj1121c knock-out mutant is impaired for flagella-mediated motility, for invasion of intestinal epithelial cells, and for persistence in the chicken intestine, clearly demonstrating that Cj1121c is essential for host colonization and virulence. Finally, we show that cj1121c is necessary for protein glycosylation by lectin Western blotting. Collectively, these results validate Cj1121c as a promising drug target and provide the means to assay for inhibitors. PMID- 16690620 TI - Beta8 integrin binds Rho GDP dissociation inhibitor-1 and activates Rac1 to inhibit mesangial cell myofibroblast differentiation. AB - Alpha(v)beta8 integrin expression is restricted primarily to kidney, brain, and placenta. Targeted alpha(v) or beta8 deletion is embryonic lethal due to defective placenta and brain angiogenesis, precluding investigation of kidney alpha(v)beta8 function. We find that kidney beta8 is localized to glomerular mesangial cells, and expression is decreased in mouse models of glomerulosclerosis, suggesting that beta8 regulates normal mesangial cell differentiation. To interrogate beta8 signaling pathways, yeast two-hybrid and co precipitation studies demonstrated beta8 interaction with Rho guanine nucleotide dissociation inhibitor-1 (GDI). Selective beta8 stimulation enhanced beta8-GDI interaction as well as Rac1 (but not RhoA) activation and lamellipodia formation. Mesangial cells from itgb8-/- mice backcrossed to a genetic background that permitted survival, or gdi-/- mice, which develop glomerulosclerosis, demonstrated RhoA (but not Rac1) activity and alpha-smooth muscle actin assembly, which characterizes mesangial cell myofibroblast transformation in renal disease. To determine whether Rac1 directly modulates RhoA-associated myofibroblast differentiation, mesangial cells were transduced with inhibitory Rac peptide fused to human immunodeficiency virus-Tat, resulting in enhanced alpha-smooth muscle actin organization. We conclude that the beta8 cytosolic tail in mesangial cells organizes a signaling complex that culminates in Rac1 activation to mediate wild-type differentiation, whereas decreased beta8 activation shifts mesangial cells toward a RhoA-dependent myofibroblast phenotype. PMID- 16690623 TI - Metabolic aspects of organogenesis in the shoot apical meristem. AB - Research over the last decade has led to tremendous advances in the characterization of the transcriptional networks involved in the initiation and maintenance of the shoot apical meristem (SAM), as well as the factors involved in the formation and development of leaves by this organ. However, one aspect of the SAM that has received rather limited attention is the fact that it is characterized by being heterotrophic, in contrast to the majority of cells and tissues immediately derived from it which rapidly undergo differentiation to form photosythetically active, autotrophic organs. This clear physiological and biochemical distinction of the SAM from the surrounding tissue raises interesting questions as to what controls the transition from heterotrophic to autotrophic growth, the nature and sequence of the metabolic events that must occur on this transition, as well as basic questions as to the potential interaction of development and metabolism in this small but essential organ of the plant. In this review, an overview is provided of present knowledge in this area, as well as some recent data that provide an insight into the potential intertwining of metabolic and developmental mechanisms during leaf initiation. PMID- 16690624 TI - Powdery mildew (Podosphaera xanthii) resistance in melon is categorized into two types based on inhibition of the infection processes. AB - Physiological races of powdery mildew (Podosphaera xanthii) cause different symptoms in eight melon lines. Infection by races 1, 2, and 5 was examined in different melon lines. After a compatible reaction, conidia germination, haustorium initiation from the germ tube, germ tube branching, and sporulation occurred within 12, 24, 48, and 120 h, respectively, and the conidia matured within 240 h. In contrast, type i and ii inhibition were identified through incompatible reactions. The germ tube and haustorium were initiated from conidia, but no germ tube branching occurred in the lines with type i resistance within 48 240 h. In type ii resistance, germ tube branching was observed within 120 h, but no sporulation was observed within 240 h. The number of fluorescing epidermal cells was higher within 24 h in type i, and within 48-120 h in type ii resistance lines than in susceptible lines. Callose accumulation around the haustorium was detected in type ii resistance lines within 48-120 h. This suggests that the rapid hypersensitive response (HR) within 24 h has an important role in the type i response, while HR and callose accumulation in the type ii response occur slowly between 48 and 120 h. Of the resistant lines, PMR 45 and WMR 29 showed a type i incompatible response; the PI 414723 response was entirely type ii; and PMR 5, PI 124112, and MR-1 showed different responses depending on the race. Therefore, the two types of incompatible responses were intermixed in the same germplasm. PMID- 16690625 TI - Early events in the perception of lipopolysaccharides in the brown alga Laminaria digitata include an oxidative burst and activation of fatty acid oxidation cascades. AB - This study provides evidence that bacterial lipopolysaccharides can be strong triggers of early events of defence reactions in the brown algal kelp Laminaria digitata, constituting the first report of a biological activity of this class of macromolecules in a marine alga. The early events include an oxidative burst, release of free saturated and unsaturated fatty acids (FFAs) and accumulation of oxylipins such as 13-hydroxyoctadecatrienoic acid and 15-hydroxyeicosapentaenoic acid. The formation of reactive oxygen species can be inhibited by diphenylene iodonium, suggesting that the source is an NAD(P)H oxidase and is similar to the oxidative burst in neutrophils and terrestrial plants. In addition and besides triggering an oxidative burst, the hypolipidemic drug clofibrate also induces the release of FFAs, to a lesser extent than lipopolysaccharides, but it does not induce oxylipin production. Other strong inducers of the oxidative burst in Laminaria such as oligoguluronates could not induce the release of FFAs nor oxylipin production. These results suggest that different signalling pathways are involved in the induction of the oxidative burst and oxylipin production. PMID- 16690626 TI - The effect of root pressurization on water relations, shoot growth, and leaf gas exchange of peach (Prunus persica) trees on rootstocks with differing growth potential and hydraulic conductance. AB - It is well known that rootstocks can have an effect on the vegetative growth and development of the tree; however, there has been no clear explanation about the physiological mechanism involved in this phenomenon. Evidence indicates that the rootstock effects on tree vegetative growth may be related to hydraulic limitations of the rootstock. The objective of these experiments was to investigate the shoot growth, water potential, and gas exchange of peach trees on different rootstocks in response to manipulations of water relations of trees on rootstocks that differ in root hydraulic conductance. Tree water relations were manipulated by applying different amounts of pneumatic pressure on the root system and then relative shoot extension growth rate, tree transpiration rate, leaf water potential, leaf conductance, leaf transpiration, and net CO(2) exchange rate responses were measured. Root pressurization increased leaf water potential, relative shoot extension growth rate, leaf conductance, leaf transpiration, and net CO(2) exchange rates of trees on both vigorous and dwarfing rootstocks. There was a significant positive linear correlation between applied pneumatic pressure and tree transpiration rate and leaf water potential. Leaf conductance, transpiration rate, and net CO(2) exchange rate as well as relative shoot extension growth rates were also positively correlated with the applied pneumatic pressure on the root system. These relationships were consistent across both vigorous and size-controlling rootstocks, indicating that rootstock hydraulic limitation may be directly involved in the vegetative growth control of peach trees. PMID- 16690627 TI - Metabolic engineering of rice with soybean isoflavone synthase for promoting nodulation gene expression in rhizobia. AB - Isoflavonoids are derived from a flavonone intermediate, naringenin, that is ubiquitously present in plants, and play a critical role in plant development and defence response. Isoflavonoids secreted by the legumes also play an important role in promoting the formation of nitrogen-fixing nodules by symbiotic rhizobia. In these plants, the key enzyme that redirects phenylpropanoid pathway intermediates from flavonoids to isoflavonoids is the cytochrome P450 mono oxygenase, isoflavone synthase. In an effort to develop a rice variety possessing the ability to induce nodulation (nod) genes in rhizobia, the IFS gene from soybean was incorporated into rice (Oryza sativa L. cv. Murasaki R86) under the control of the 35S promoter. The presence of IFS in transgenic rice was confirmed by PCR and Southern blot analysis. Analyses of the 35S-IFS transgenic lines demonstrated that the expression of the IFS gene led to the production of the isoflavone genistein in rice tissues. These results showed that the soybean IFS gene-expressed enzyme is active in the R86 rice plant, and that the naringenin intermediate of the anthocyanin pathway is available as a substrate for the introduced foreign enzyme. The genistein produced in rice cells was present in a glycoside form, indicating that endogenous glycosyltransferases were capable of recognizing genistein as a substrate. Studies with rhizobia demonstrated that the expression of isoflavone synthase confers rice plants with the ability to produce flavonoids that are able to induce nod gene expression, albeit to varied degrees, in different rhizobia. PMID- 16690628 TI - A formal comparison of the practice of assisted reproductive technologies between Europe and the USA. AB - In this study, we compared pregnancy and delivery outcomes after the utilization of assisted reproductive technologies (ART) in Europe and the United States (US). ART outcomes were compared between Europe and the US for the year 2001, based on formal reports published by the European Society for Human Reproduction and Embryology (ESHRE) and the Center for Disease Control and Prevention (CDC) in collaboration with the American Society for Reproductive Medicine (ASRM) and Society for ART (SART). Europe utilizes ART at approximately twice the rate of the US (P < 0.001). United States patients showed a significantly decreased likelihood of reaching oocyte retrieval (P < 0.001) and embryo transfer (P < 0.001). Despite this lower chance of reaching oocyte retrievals and embryo transfers, US patients experienced significantly higher clinical pregnancy rates (P < 0.001) and delivery rates per started cycle (P < 0.001) than European patients. Amongst patients reaching oocyte retrieval, the difference in clinical pregnancy rates and live birth rates was even more pronounced in favour of the US. However, US patients received significantly more embryos per embryo transfer (P < 0.001) and experienced a significantly higher multiple pregnancy rate (P < 0.001). Significant differences in favour of US patients in pregnancy rates and live birth rates were also observed for frozen embryo cycles and oocyte donation cycles, where the difference was most pronounced. The better pregnancy and live birth outcomes in the US are not explainable by the transfer of larger embryo numbers alone. PMID- 16690629 TI - A puzzling telemetry tracing. PMID- 16690630 TI - Modelling the health benefits and economic implications of implanting dual chamber vs. single-chamber ventricular pacemakers in the UK. AB - AIMS: To estimate the consequences of managing bradycardia due to sinoatrial node disease or atrioventricular block with dual-chamber vs. single-chamber ventricular pacemakers. METHODS AND RESULTS: A discrete-event simulation was conducted to predict outcomes over 5 years. Patients could develop post-operative complications, clinically relevant pacemaker syndrome leading to replacement of single-chamber with dual-chamber, atrial fibrillation (AF; which if chronic might require anticoagulants) or stroke. Survival, quality-adjusted life years (QALYs), complications, and associated direct medical costs were estimated (2003 British Pounds pounds sterling). Identical patients were simulated after receiving a single-chamber device or a more expensive dual-chamber pacemaker. Probabilities of conditions were obtained from clinical trials. Benefits were discounted at 1.5% and costs at 6%. Post-operative complications increased from 6.4% with single-chamber to 7.7% with dual-chamber but AF decreased (22 vs. 18%) as did clinically relevant pacemaker symptoms (16.8 vs. 0%). Approximately 4300 pounds sterling were accrued per patient over 5 years. Additional health benefits with dual-chamber are achieved at a mean net cost of 43 pounds sterling per patient, leading to 0.09 QALY with a cost-effectiveness ratio of 477 pounds sterling/QALY. CONCLUSION: Implanting the costlier device increases the cost of the initial operation; however, this is expected to be offset by a reduction in costs associated with re-operations and AF. PMID- 16690631 TI - Patient and staff radiation dosimetry during cardiac electrophysiology studies and catheter ablation procedures: a comprehensive analysis. AB - AIMS: To perform a comprehensive analysis of all aspects of patient and in-room personnel radiation dosimetry in interventional electrophysiology. METHODS AND RESULTS: Measurements were performed during 19 diagnostic electrophysiology studies and 24 catheter ablations. Kerma-area product and exposure time values were 48.7 (6.4-230) Gy cm2 and 25.5 (4.4-79.2) min for ablation, and 12.5 (4.5 117.2) Gy cm2 and 4.5 (1.2-31) min for diagnostic studies, respectively. Patient effective doses were 15.2 (2.1-59.6) mSv for ablation and 3.2 (1.3-23.9) mSv for diagnostic procedures. Radiation risk to the patient was estimated to be up to eight cases of fatal cancer in 10,000 procedures. The risk of development of fatal cancer was less than 3x10(-6) per procedure to the primary operator. The risk for the nurse and technician was much lower. The dose per procedure for the primary operator was 7.1 microGy at the eyes, 0.79 microGy at the chest under the lead apron, 13.68 microGy at the chest over the apron, 3.82 microGy at the thyroid, 17.76 microGy at the left hand, and 12.11 microGy at the left knee. CONCLUSION: As far as radiation exposure is concerned, electrophysiology studies followed by radiofrequency ablation are safe procedures for both patient and personnel when performed in catheterization laboratories with modern equipment, experienced operators, and standard safety precautions. PMID- 16690632 TI - Long-term follow-up of the signal-averaged ECG in arrhythmogenic right ventricular cardiomyopathy: correlation with arrhythmic events and echocardiographic findings. AB - AIMS: The aims of our study were to evaluate late potential changes during long term follow-up in patients with arrhythmogenic right ventricular cardiomyopathy (ARVC) and to correlate these results with echocardiographic findings and sustained ventricular tachycardia (VT) occurrence. METHODS AND RESULTS: We studied 31 patients (22 males and 9 females; mean age 29+/-16) during 8 years of follow-up by signal-averaged ECG (SAECG) and echocardiography. Ten subjects experienced episodes of sustained VT. During follow-up, all the SAECG parameters showed a progressive significant increase in late potentials. In contrast, echocardiographic indices did not show evidence of relevant modifications. Patients with sustained VT were characterized by significantly lower left and right ventricular ejection fractions, longer values of filtered QRS at 25/40/80 250 Hz filters, and longer high-frequency low-amplitude (HFLA) signals at 25-250 Hz at baseline. The analysis of SAECG modification during follow-up indicated that only HFLA signals at 25-250 Hz increased significantly in the sustained VT group. CONCLUSION: We detected a progressive increase in delayed ventricular conduction by SAECG not associated with significant echocardiographic changes. Therefore, the conduction disturbance seems to increase independently from anatomical alterations. The baseline SAECG and echocardiographic parameters, more than their modifications during follow-up, appear to be useful in identifying patients with sustained VT. PMID- 16690633 TI - Synchronizing a multicellular system by external input: an artificial control strategy. AB - MOTIVATION: Although there are significant advances on elucidating the collective behaviors on biological organisms in recent years, the essential mechanisms by which the collective rhythms arise remain to be fully understood, and further how to synchronize multicellular networks by artificial control strategy has not yet been well explored. RESULTS: A control strategy is developed to synchronize gene regulatory networks in a multicellular system when spontaneous synchronization cannot be achieved. We first construct an impulsive control system to model the process of periodically injecting coupling substances with constant or random impulsive control amounts into the common extracellular medium, and further study its effects on the dynamics of individual cells. We derive the threshold of synchronization induced by the periodic substance input. Therefore, we can synchronize the multicellular network to a specific collective behavior by changing the frequency and amplitude of the periodic stimuli. Moreover, a two stage scheme is proposed to facilitate the synchronization in this paper. We show that the presence of the external input may also initiate different dynamics. The multicellular network of coupled repressilators is used to show the effectiveness of the proposed method. The results not only provide a perspective to understand the interactions between external stimuli and intrinsic physiological rhythms, but also may lead to development of realistic artificial control strategy and medical therapy. AVAILABILITY: CONTACT: aihara@sat.t.u-tokyo.ac.jp. PMID- 16690634 TI - SCARNA: fast and accurate structural alignment of RNA sequences by matching fixed length stem fragments. AB - MOTIVATION: The functions of non-coding RNAs are strongly related to their secondary structures, but it is known that a secondary structure prediction of a single sequence is not reliable. Therefore, we have to collect similar RNA sequences with a common secondary structure for the analyses of a new non-coding RNA without knowing the exact secondary structure itself. Therefore, the sequence comparison in searching similar RNAs should consider not only their sequence similarities but also their potential secondary structures. Sankoff's algorithm predicts the common secondary structures of the sequences, but it is computationally too expensive to apply to large-scale analyses. Because we often want to compare a large number of cDNA sequences or to search similar RNAs in the whole genome sequences, much faster algorithms are required. RESULTS: We propose a new method of comparing RNA sequences based on the structural alignments of the fixed-length fragments of the stem candidates. The implemented software, SCARNA (Stem Candidate Aligner for RNAs), is fast enough to apply to the long sequences in the large-scale analyses. The accuracy of the alignments is better or comparable with the much slower existing algorithms. AVAILABILITY: The web server of SCARNA with graphical structural alignment viewer is available at http://www.scarna.org/. PMID- 16690635 TI - Motor plasticity in a juggling task in older adults-a developmental study. AB - OBJECTIVE: to examine the plasticity of motor performance in old age. Older adults were instructed and trained in a juggling task and their performances were compared, first, within the group of older adults and, second, with the performances of children, youths and younger adults. SUBJECTS: older adults, children, youths and younger adults (n = 1,206, range 6-89 years). METHODS: participants were asked to learn a juggling task. Performance was tested before semantic instruction (pre-test 1), after semantic instruction (pre-test 2) and after 6 days of juggling practice (post-test). None of the participants had prior experiences in juggling. Results were analysed using repeated measure analysis of variance (ANOVA). RESULTS: older adults showed a clear improvement in juggling performance after instruction and after six training sessions. On average, they reached performances comparable with those of children aged between 10 and 14 years, and with those of younger adults aged between 30 and 59 years. Only youths and younger adults aged between 15 and 29 years showed significantly higher performances at baseline, after instruction and after training. CONCLUSIONS: older adults exhibit high reserve capacity, that is, a potential for learning 'new' motor skills. PMID- 16690636 TI - Is grip strength associated with health-related quality of life? Findings from the Hertfordshire Cohort Study. AB - OBJECTIVE: to investigate the relationship between grip strength and health related quality of life (HRQoL). DESIGN: cross-sectional survey within a cohort study design. SETTING: the county of Hertfordshire in the UK. PARTICIPANTS: a total of 2,987 community-dwelling men and women aged 59-73 years of age. MEASUREMENTS: grip strength was used as a marker of sarcopaenia and measured using a Jamar dynamometer. HRQoL was assessed using the eight domain scores of the Short Form-36 (SF-36) questionnaire, and subjects in the lowest sex-specific fifth of the distribution were classified as having 'poor' status for each domain. RESULTS: men and women with lower grip strength were significantly more likely to report a poor as opposed to excellent to fair overall opinion of their general health (GH) [odds ratio (OR) per kilogram decrease in grip strength = 1.13, 95% CI = 1.06-1.19, P < 0.001 in men, 1.13, 95% CI = 1.07-1.20, P < 0.001 in women]. Among men, after adjustment for age, size, physical activity and known co-morbidity, decreased grip strength was associated with increased prevalence of poor SF-36 scores for the physical functioning (PF) (OR per kilogram decrease in grip strength = 1.03, 95% CI = 1.01-1.06, P = 0.007) and GH domains (OR = 1.03, 95% CI = 1.01-1.05, P = 0.01). Similar associations were seen in women. CONCLUSIONS: our findings suggest that lower grip strength is associated with reduced HRQoL in older men and women. This does not appear to be explained by age, size, physical activity or co-morbidity and may reflect the link between sarcopaenia and generalised frailty. Individuals with sarcopaenia may benefit from interventions to improve muscle mass and strength before the onset of chronic disorders usually associated with impaired HRQoL. PMID- 16690637 TI - Prevalence, duration and indications for prescribing of antidepressants in primary care. PMID- 16690638 TI - Women with wrist fractures are at increased risk for future fractures because of both skeletal and non-skeletal risk factors. PMID- 16690639 TI - Population pharmacokinetics and dosing regimen design of milrinone in preterm infants. AB - AIMS: To define the pharmacokinetics of milrinone in very preterm infants and determine an optimal dose regimen to prevent low systemic blood flow in the first 12 h after birth. METHODS: A prospective open-labelled, dose-escalation pharmacokinetic study was undertaken in two stages. In stage one, infants received milrinone at 0.25 microg/kg/min (n = 8) and 0.5 microg/kg/min (n = 11) infused from 3 to 24 h of age. Infants contributed 4-5 blood samples for concentration-time data which were analysed using a population modelling approach. A simulation study was used to explore the optimal dosing regimen to achieve target milrinone concentrations (180-300 ng/ml). This milrinone regimen was evaluated in stage two (n = 10). RESULTS: Infants (n = 29) born before 29 weeks gestation were enrolled. Milrinone pharmacokinetics were described using a one-compartment model with first-order elimination rate, with a population mean clearance (CV%) of 35 ml/h (24%) and volume of distribution of 512 ml (21%) and estimated half-life of 10 h. The 0.25 and 0.5 microg/kg/min dosage regimens did not achieve optimal milrinone concentration-time profiles to prevent early low systemic blood flow. Simulation studies predicted a loading infusion (0.75 microg/kg/min for 3 h) followed by maintenance infusion (0.2 microg/kg/min until 18 h of age) would provide an optimal milrinone concentration profile. This was confirmed in stage two of the study. CONCLUSION: Population pharmacokinetic modelling in the preterm infant has established an optimal dose regimen for milrinone that increases the likelihood of achieving therapeutic aims and highlights the importance of pharmacokinetic studies in neonatal clinical pharmacology. PMID- 16690640 TI - Randomised trial of high frequency oscillatory ventilation or conventional ventilation in babies of gestational age 28 weeks or less: respiratory and neurological outcomes at 2 years. AB - BACKGROUND: The long term outcome of children entered into neonatal trials of high frequency oscillatory ventilation (HFOV) or conventional ventilation (CV) has been rarely studied. OBJECTIVE: To evaluate respiratory and neurodevelopmental outcomes for children entered into the United Kingdom Oscillation Study, which was designed to evaluate these outcomes. METHODS: Surviving infants were followed until 2 years of age corrected for prematurity. Study forms were completed by local paediatricians at routine assessments, and parents were asked to complete a validated neurodevelopmental questionnaire. RESULTS: Paediatricians' forms were returned for 73% of the 585 surviving infants. Respiratory symptoms were common in all infants, and 41% had received inhaled medication. Mode of ventilation had no effect on frequency of any symptoms. At 24 months of age, severe neurodevelopmental disability was present in 9% and other disabilities in 38% of children, but the prevalence of disability was similar in children who received HFOV or CV (relative risk 0.93; 95% confidence interval 0.74 to 1.16). The prevalence of disability did not vary by gestational age, but boys were more likely to have overall disability. Developmental scores were unaffected by mode of ventilation (relative risk 1.13; 95% confidence interval 0.78 to 1.63) and were lower in infants born before 26 weeks gestation compared with babies born at 26-28 weeks. CONCLUSIONS: Initial mode of ventilation in very preterm infants has no impact on respiratory or neurodevelopmental morbidity at 2 years. HFOV and CV appear equally effective for the early treatment of respiratory distress syndrome. PMID- 16690641 TI - Clinical effectiveness of treatment with hyperbaric oxygen for neonatal hypoxic ischaemic encephalopathy: systematic review of Chinese literature. AB - OBJECTIVES: To investigate the clinical effectiveness of treatment with hyperbaric oxygen for neonates with hypoxic-ischaemic encephalopathy. This treatment is frequently used in China but much less often in the West. DATA SOURCES: Western (Cochrane controlled trials register and database of systematic reviews, Medline, Embase, CINAHL, and HealthSTAR) and Chinese (China Hospital Digital Library, Chinese Medical Journal Network) databases and hand search of Chinese journals. No language restrictions. REVIEW METHODS: Randomised or quasi randomised controlled trials of treatment with hyperbaric oxygen compared with "usual care" in term neonates with hypoxic-ischaemic encephalopathy. Outcomes included mortality and long term neurological sequelae. Standardised forms were used to extract and compare data. Criteria of York Centre for Reviews and Dissemination were used to assess quality. Analysis was mainly qualitative but included meta-analysis. RESULTS: 20 trials were found, mainly from Chinese sources. The reporting quality of trials was poor by Western (CONSORT) standards. Treatment with hyperbaric oxygen had better outcomes than the comparator in almost all trials. The odds ratios of the meta-analyses were 0.26 (95% confidence interval 0.14 to 0.46) for mortality and 0.41 (0.27 to 0.61) for neurological sequelae. CONCLUSION: Treatment with hyperbaric oxygen possibly reduces mortality and neurological sequelae in term neonates with hypoxic-ischaemic encephalopathy. Because of the poor quality of reporting in all trials and the possibility of publication bias, an adequately powered, high quality randomised controlled trial is needed to investigate these findings. The Chinese medical literature may be a rich source of evidence to inform clinical practice and other systematic reviews. PMID- 16690642 TI - Family size, fertility preferences, and sex ratio in China in the era of the one child family policy: results from national family planning and reproductive health survey. AB - OBJECTIVES: To examine the impact of the one child family policy in China on fertility, preferred family size, and sex ratio. DESIGN: Secondary analysis of data from the Chinese cross sectional national family planning and reproductive health survey, 2001. Interviews of representative sample of women aged 15-49. RESULTS: Data were obtained from 39,585 women, with a total of 73,202 pregnancies and 56,830 live births. The average fertility rate in women over 35 (n = 17,078) was 1.94 (2.1 in rural areas and 1.4 in urban areas) and for women under 35 (n = 11,543) 1.73 (1.25 and 1.79). Smaller families were associated with younger age, higher level of education, and living in an urban area. The male to female ratio was 1.15 and rose from 1.11 in 1980-9 to 1.23 for 1996-2001. Most women wanted small families: 35% preferred one child and 57% preferred two. CONCLUSION: Since the one child family policy began, the total birth rate and preferred family size have decreased, and a gross imbalance in the sex ratio has emerged. PMID- 16690643 TI - Diabetes and lipid lowering: where are we? PMID- 16690644 TI - Cerebral embolism and Alzheimer's disease. PMID- 16690645 TI - Kidneys for transplant. PMID- 16690646 TI - HPV vaccine and adolescents' sexual activity. PMID- 16690647 TI - An international standard for disclosure of clinical trial information. PMID- 16690649 TI - Adverse drug reactions result in 250,000 UK admissions a year. PMID- 16690650 TI - GMC must consider case against paediatricians who suspected parents of fabricating child's illness. PMID- 16690651 TI - Belgium to tighten drug advertising laws. PMID- 16690652 TI - Chairwoman of Shipman inquiry protests at lack of action. PMID- 16690653 TI - One in four African children have to work. PMID- 16690654 TI - Thousands of Ugandans with AIDS don't get free drugs. PMID- 16690655 TI - Zuma's trial leaves confused messages on AIDS. PMID- 16690656 TI - EU companies can export generic versions of patented drugs to poor countries. PMID- 16690658 TI - Cost of out of hours care was 22% higher than predicted in England. PMID- 16690657 TI - Cancer charity is to "borrow" candidate drugs shelved by companies. PMID- 16690661 TI - Bush administration announces plan to combat bird flu. PMID- 16690663 TI - Couple is given go-ahead to use embryo selection to help existing child. PMID- 16690665 TI - Doctors' career structure to include management training. PMID- 16690669 TI - Improving the quality and clinical relevance of diagnostic studies. PMID- 16690670 TI - Sodium in soluble paracetamol may be linked to raised blood pressure. PMID- 16690671 TI - Diagnosis and management of dysmenorrhoea. PMID- 16690672 TI - Anomalous origin of left coronary artery in young athletes with syncope. PMID- 16690673 TI - ABC of chronic obstructive pulmonary disease. Definition, epidemiology, and risk factors. PMID- 16690676 TI - Informed choice for screening: implications for evaluation. PMID- 16690677 TI - Common susceptibility genes for cancer: search for the end of the rainbow. PMID- 16690678 TI - Confounders in chronic stress at work and metabolic syndrome. PMID- 16690679 TI - Hamstring v patellar tendon autografts: when to operate is important. PMID- 16690680 TI - Hamstring v patellar tendon autografts: a patient's and clinician's perspective. PMID- 16690681 TI - Psychological treatment centres: the case against. PMID- 16690682 TI - Depression should be managed like a chronic disease: what exactly is "depression"? PMID- 16690683 TI - Depression should be managed like a chronic disease: myth of 15% suicide rate was promulgated again. PMID- 16690684 TI - Research for profit. PMID- 16690685 TI - Psychological treatment centres: Layard's vision has already started. PMID- 16690686 TI - Psychological treatment centres: may be less effective than suggested. PMID- 16690687 TI - Psychological treatment centres: quality of psychological treatment is important. PMID- 16690688 TI - Selecting medical students: a struggler responds. PMID- 16690689 TI - Selecting medical students: evidence based admissions procedures for medical students are being tested. PMID- 16690690 TI - Vacuums and maggots in orthopaedic surgery. PMID- 16690691 TI - Costs and quality of life of patients with multiple sclerosis in Europe. AB - OBJECTIVE: To assess overall resource consumption, work capacity and quality of life of patients with multiple sclerosis in nine European countries. METHODS: Information on resource consumption related to multiple sclerosis, informal care by relatives, productivity losses and overall quality of life (utility) was collected with a standardised pre-tested questionnaire from 13,186 patients enrolled in national multiple sclerosis societies or followed up in neurology clinics. Information on disease included disease duration, self-assessed disease severity and relapses. Mean annual costs per patient (Euro, 2005) were estimated from the societal perspective. RESULTS: The mean age ranged from 45.1 to 53.4 years, and all levels of disease severity were represented. Between 16% and 29% of patients reported experiencing a relapse in the 3 months preceding data collection. The proportion of patients in early retirement because of multiple sclerosis ranged from 33% to 45%. The use of direct medical resources (eg, hospitalisation, consultations and drugs) varied considerably across countries, whereas the use of non-medical resources (eg, walking sticks, wheel chairs, modifications to house and car) and services (eg, home care and transportation) was comparable. Informal care use was highly correlated with disease severity, but was further influenced by healthcare systems and family structure. All types of costs increased with worsening disease. The total mean annual costs per patient (adjusted for gross domestic product purchasing power) were estimated at Euro 18,000 for mild disease (Expanded Disability Status Scale (EDSS) <4.0), Euro 36,500 for moderate disease (EDSS 4.0-6.5) and Euro 62,000 for severe disease (EDSS >7.0). Utility was similar across countries at around 0.70 for a patient with an EDSS of 2.0 and around 0.45 for a patient with an EDSS of 6.5. Intangible costs were estimated at around Euro 13,000 per patient. PMID- 16690692 TI - Arnold-Chiari-II malformation and cognitive functioning in spina bifida. AB - Spina bifida is a multifaceted neurological condition with complex neuropsychological sequelae. The cognitive outcome in spina bifida has frequently been attributed to the severity of the hydrocephalus. However, because of complex neuropathology, the influence of hydrocephalus alone does not sufficiently explain the deficits in the cognitive profile in spina bifida. To date, little is known of the role of Arnold-Chiari-II malformation (ACM) in the cognitive profile of these patients. Aim of the current study is to delineate the specific contribution of the ACM in spina bifida by comparing children with ACM and those without ACM. 46 children between 6 and 15 years of age underwent a neuropsychological assessment covering intelligence and a wide range of cognitive functions, such as visuo-motor processing, attention, memory, word fluency and speed of information processing. Comparisons were made between patients with ACM (ACM+) and those without ACM (ACM-); all children with ACM+ also had hydrocephalus. Confounding effects of global cognitive impairment were excluded, such that groups were matched on verbal IQ. Because of complex neuropathology, which is inherent to spina bifida, the method applied was based on a comparison of cognitive profiles of the study group with profiles of patients with cerebellar damage and hydrocephalus found in the literature. Impaired visual analysis and synthesis, verbal memory, and verbal fluency, even after correction for global cognitive impairment, were observed in children with ACM. The hypothesis that in addition to impairment in visual analysis and synthesis, which are related to both hydrocephalus and ACM, specific deficiencies in verbal memory and fluency may be attributed to ACM is supported. PMID- 16690693 TI - Effects of focal hand dystonia on visually guided and internally guided force control. AB - BACKGROUND: A fundamental feature underlying many movement disorders is increased variability in the motor response. Despite abnormalities of grip force control in people with dystonia, it is not clear whether dystonia is also associated with increased variability in force output and whether force variability in dystonia is affected by the presence or absence of visual feedback. OBJECTIVE: To examine force variability in 16 patients with writer's cramp and 16 matched controls. METHODS: The variability of force output at the wrist under conditions of both vision and no vision was examined. The underlying frequency structure of the force signal was also compared across groups. Participants produced isometric wrist flexion to targets at 25% and 50% of their maximum voluntary contraction strength under conditions of both vision and no vision. RESULTS: Similar levels of force variability were observed in patients with dystonia and controls at the lower force levels, but patients with dystonia were less variable in their force output than controls at the higher force level. This reduction in variability in people with dystonia at 50% maximum voluntary contraction was not affected by vision. Although a similar dominant frequency in force output was observed in people with dystonia and controls, a reduced variability in the group with dystonia at the higher force level was due to reduced power in the 0-4-Hz frequency bin. CONCLUSIONS: The first evidence of a movement disorder with reduced variability is provided. The findings are compatible with a model of dystonia, which includes reduced cortical activation in response to sensory input from the periphery and reduced flexibility in motor output. PMID- 16690694 TI - Is hypertension a more frequent risk factor for deep than for lobar supratentorial intracerebral haemorrhage? AB - OBJECTIVE: To determine whether evidence from observational studies supports the widely held belief that hypertension is more commonly a risk factor for deep than for lobar supratentorial intracerebral haemorrhage. METHODS: Studies comparing the frequency of hypertension as a risk factor for deep versus lobar supratentorial intracerebral haemorrhage, excluding haemorrhages with identified secondary causes, were identified and subjected to a meta-analysis. The effects of predefined methodological quality criteria on the results were assessed and other sources of bias were considered. RESULTS: The pooled result from all 28 included studies (about 4000 patients) found hypertension to be about twice as common in patients with deep as in those with lobar haemorrhage (odds ratio (OR) 2.10, 95% confidence interval (95% CI) 1.82 to 2.42), but there was significant heterogeneity between studies. The pooled OR was less extreme for studies that used a pre-stroke definition of hypertension, were population based or included first-ever strokes only. In the three studies meeting all criteria (601 patients), deep haemorrhage was associated with a smaller, statistically significant excess of hypertension (OR 1.50, 95% CI 1.09 to 2.07). The OR for studies confined to younger patients seemed to be more extreme (12.32, 95% CI 6.13 to 24.77), but none of these studies fulfilled our methodological quality criteria. Additional, unquantified sources of bias included uncertainty about whether those doctors reporting brain scans were blind to hypertension status, uncertain reliability of the classification of haemorrhage location and variable rates of investigation for secondary causes. CONCLUSIONS: An excess of hypertension was found in patients with deep versus lobar intracerebral haemorrhages without an identified secondary cause, but this may be due to residual, unquantified methodological biases. PMID- 16690695 TI - Bilateral grey-matter increase in the putamen in primary blepharospasm. AB - BACKGROUND: Primary blepharospasm is a focal dystonia characterised by excessive involuntary closure of the eyelids. The pathophysiology of primary blepharospasm is unresolved. AIM: To pinpoint grey-matter changes that are associated with primary blepharospasm. METHODS: 16 right-handed patients with primary blepharospasm (mean age 67.4 (SD 4.3) years; 12 women) were compared with 16 healthy volunteers matched for sex and age. High-resolution T1-weighted magnetic resonance imaging of each participant was obtained and analysed by voxel-based morphometry, a method to detect regionally specific differences in grey matter between patients and control group. To evaluate whether the identified grey matter changes were correlated with the duration of primary blepharospasm or botulinum neurotoxin treatment (BoNT), separate regression analyses were carried out. RESULTS: In patients with primary blepharospasm, grey-matter increase in the putamina was observed, whereas regression analyses did not indicate a correlation between grey-matter increases and the duration of primary blepharospasm or BoNT. Grey-matter decrease was detected in the left inferior parietal lobule; here regression analyses of grey-matter decrease showed a significant (p = 0.013) correlation of grey-matter decrease with the duration of BoNT. CONCLUSIONS: The data suggest structural changes in primary blepharospasm and point to a crucial role of the putamen for the pathophysiology of this focal dystonia. PMID- 16690696 TI - Cognitive outcome 5 years after bilateral chronic stimulation of subthalamic nucleus in patients with Parkinson's disease. AB - AIM: To assess the long-term cognitive and behavioural outcome after bilateral deep brain stimulation (DBS) of the subthalamic nucleus (STN) in patients affected by Parkinson's disease, with a 5-year follow-up after surgery. METHODS: 11 patients with Parkinson's disease treated by bilateral DBS of STN underwent cognitive and behavioural assessments before implantation, and 1 and 5 years after surgery. Postoperative cognitive assessments were carried out with stimulators turned on. RESULTS: A year after surgery, there was a marginally significant decline on a letter verbal fluency task (p = 0.045) and a significant improvement on Mini-Mental State Examination (p = 0.009). 5 years after surgery, a significant decline was observed on a letter verbal fluency task (p = 0.007) and an abstract reasoning task (p = 0.009), namely Raven's Progressive Matrices 1947. No significant postoperative change was observed on other cognitive variables. No patient developed dementia 5 years after surgery. A few days after the implantation, two patients developed transient manic symptoms with hypersexuality and one patient developed persistent apathy. CONCLUSION: The decline of verbal fluency observed 5 years after implantation for DBS in STN did not have a clinically meaningful effect on daily living activities in our patients with Parkinson's disease. As no patient developed global cognitive deterioration in our sample, these findings suggest that DBS of STN is associated with a low cognitive and behavioural morbidity over a 5-year follow-up, when selection criteria for neurosurgery are strict. PMID- 16690697 TI - Life-threatening hypernatraemic dehydration in breastfed babies. AB - We describe five babies, who were exclusively breast fed, with life-threatening complications of hypernatraemic dehydration secondary to inadequate breast feeding. An increased awareness among health professionals is required so that this potentially devastating condition can be prevented. PMID- 16690698 TI - Systematic review of effectiveness of bisphosphonates in treatment of low bone mineral density and fragility fractures in juvenile idiopathic arthritis. AB - AIMS: To evaluate the currently available evidence for the effectiveness of bisphosphonates in children with low bone mineral density (BMD) and fragility fractures associated with juvenile idiopathic arthritis (JIA), and the safety of bisphosphonates in JIA and other conditions. METHODS: Literature databases were searched using a structured search strategy. The effectiveness review included any studies of children with JIA treated with bisphosphonates. The safety review also included studies of osteogenesis imperfecta. Quantitative data analysis was not undertaken because of the heterogeneity of the studies; findings were summarised using tables and narrative synthesis. RESULTS: Ninety four studies were identified. Sixteen studies (78 JIA children) were included in the effectiveness review: one randomised controlled trial, three controlled cohort studies, 11 case series, and one case report. At baseline, children had low BMD below the expected values for age and sex matched children. In all studies, treatment with bisphosphonates increased BMD compared with baseline: the mean percentage increase in spine BMD ranged from 4.5% to 19.1%. Overall, studies were heterogeneous and of variable quality. A total of 59 papers were included in the safety review; treatment durations were up to three years. The most common side effect was a flu-like reaction with intravenous treatment. This occurred during the first infusion and was transient; the symptoms were managed with paracetamol and did not occur during subsequent cycles. CONCLUSIONS: Bisphosphonates are a promising treatment for low BMD and fragility fractures in children with JIA. However, the quality of the current evidence is variable and better studies are needed to more clearly assess their role. PMID- 16690699 TI - The incidence of inherited metabolic disorders in the West Midlands, UK. AB - BACKGROUND: Inherited metabolic disorders (IMDs) are a heterogeneous group of genetic conditions mostly occurring in childhood. They are individually rare but collectively numerous, causing substantial morbidity and mortality. AIMS: To obtain up-to-date estimates of the birth prevalence of IMDs in an ethnically diverse British population and to compare these estimates with those of other published population-based studies. METHODS: Retrospective data from the West Midlands Regional Diagnostic Laboratory for Inherited Metabolic Disorders (Birmingham, UK) for the 5 years (1999-2003) were examined. The West Midlands population of 5.2 million is approximately 10% of the UK population. Approximately 11% of the population of the region is from black and ethnic minority groups compared with approximately 8% for the the UK. RESULTS: The overall birth prevalence was 1 in 784 live births (95% confidence interval (CI) 619 to 970), based on a total of 396 new cases. The most frequent diagnoses were mitochondrial disorders (1 in 4929; 95% CI 2776 to 8953), lysosomal storage disorders (1 in 5175; 95% CI 2874 to 9551), amino acid disorders excluding phenylketonuria (1 in 5354; 95% CI 2943 to 9990) and organic acid disorders (1 in 7962; 95% CI 3837 to 17 301). Most of the diagnoses (72%) were made by the age of 15 years and one-third by the age of 1 year. CONCLUSIONS: These results are similar to those of the comparison studies, although the overall birth prevalence is higher in this study. This is probably due to the effects of ethnicity and consanguinity and increasing ascertainment. This study provides useful epidemiological information for those planning and providing services for patients with IMDs, including newborn screening, in the UK and similar populations. PMID- 16690700 TI - Exercising women throw a wrench in the gears of the AMPK-lipid oxidation link. PMID- 16690701 TI - Ca2+-calmodulin-dependent protein kinase expression and signalling in skeletal muscle during exercise. AB - Ca2+ signalling is proposed to play an important role in skeletal muscle function during exercise. Here, we examined the expression of multifunctional Ca2+ calmodulin-dependent protein kinases (CaMK) in human skeletal muscle and show that CaMKII and CaMKK, but not CaMKI or CaMKIV, are expressed. Furthermore, the effect of exercise duration and intensity on skeletal muscle CaMKII activity and phosphorylation of downstream targets was examined. Eight healthy men exercised at approximately 67% of peak pulmonary O2 uptake(VO2peak) with muscle samples taken at rest and after 1, 10, 30, 60 and 90 min of exercise. Ten other men exercised for three consecutive 10 min bouts at 35%, 60% and 85% VO2peak with muscle samples taken at rest, at the end of each interval and 30 min post exercise. There was a rapid and transient increase in autonomous CaMKII activity and CaMKII phosphorylation at Thr287 in skeletal muscle during exercise. Furthermore, the phosphorylation of phospholamban (PLN) at Thr17, which was identified as a CaMKII substrate in skeletal muscle, was rapidly (< 1 min) increased by exercise, and remained phosphorylated 5-fold above basal level during 90 min of exercise. The phosphorylation of serum response factor at Ser103, a putative CaMKII substrate, was higher after 30 min of exercise. PLN phosphorylation at Thr17 was higher with increasing exercise intensities. These data indicate that CaMKII is the major multifunctional CaMK in skeletal muscle and its activation occurs rapidly and is sustained during continuous exercise, with the activation being greater during intense exercise. PMID- 16690702 TI - Ionic mechanisms of autorhythmic firing in rat cerebellar Golgi cells. AB - Although Golgi cells (GoCs), the main type of inhibitory interneuron in the cerebellar granular layer (GL), are thought to play a central role in cerebellar network function, their excitable properties have remained unexplored. GoCs fire rhythmically in vivo and in slices, but it was unclear whether this activity originated from pacemaker ionic mechanisms. We explored this issue in acute cerebellar slices from 3-week-old rats by combining loose cell-attached (LCA) and whole-cell (WC) recordings. GoCs displayed spontaneous firing at 1-10 Hz (room temperature) and 2-20 Hz (35-37 degrees C), which persisted in the presence of blockers of fast synaptic receptors and mGluR and GABAB receptors, thus behaving, in our conditions, as pacemaker neurons. ZD 7288 (20 microM), a potent hyperpolarization-activated current (Ih) blocker, slowed down pacemaker frequency. The role of subthreshold Na+ currents (INa,sub) could not be tested directly, but we observed a robust TTX-sensitive, non-inactivating Na+ current in the subthreshold voltage range. When studying repolarizing currents, we found that retigabine (5 microM), an activator of KCNQ K+ channels generating neuronal M-type K+ (IM) currents, reduced GoC excitability in the threshold region. The KCNQ channel antagonist XE991 (5 microM) did not modify firing, suggesting that GoC IM has low XE991 sensitivity. Spike repolarization was followed by an after hyperpolarization (AHP) supported by apamin-sensitive Ca2+-dependent K+ currents (I(apa)). Block of I(apa) decreased pacemaker precision without altering average frequency. We propose that feed-forward depolarization is sustained by Ih and INa,sub, and that delayed repolarizing feedback involves an IM-like current whose properties remain to be characterized. The multiple ionic mechanisms shown here to contribute to GoC pacemaking should provide the substrate for fine regulation of firing frequency and precision, thus influencing the cyclic inhibition exerted by GoCs onto the cerebellar GL. PMID- 16690703 TI - Nerve growth factor affects Ca2+ currents via the p75 receptor to enhance prolactin mRNA levels in GH3 rat pituitary cells. AB - In clonal pituitary GH(3) cells, spontaneous action potentials drive the opening of Ca(v)1 (L-type) channels, leading to Ca(2+) transients that are coupled to prolactin gene transcription. Nerve growth factor (NGF) has been shown to stimulate prolactin synthesis by GH(3) cells, but the underlying mechanisms are unknown. Here we studied whether NGF influences prolactin gene expression and Ca(2+) currents. By using RT-PCR, NGF (50 ng ml(-1)) was found to augment prolactin mRNA levels by approximately 80% when applied to GH(3) cells for 3 days. A parallel change in the prolactin content was detected by Western blotting. Both NGF-induced responses were mimicked by an agonist (Bay K 8644) and prevented by a blocker (nimodipine) of L-type channels. In whole-cell patch-clamp experiments, NGF enhanced the L-type Ca(2+) current by approximately 2-fold within 60 min. This effect reversed quickly upon growth factor withdrawal, but was maintained for days in the continued presence of NGF. In addition, chronic treatment (>or= 24 h) with NGF amplified the T-type current, which flows through Ca(v)3 channels and is thought to support pacemaking activity. Thus, NGF probably increases the amount of Ca(2+) that enters per action potential and may also induce a late increase in spike frequency. MC192, a specific antibody for the p75 neurotrophin receptor, but not tyrosine kinase inhibitors (K252a and lavendustin A), blocked the effects of NGF on Ca(2+) currents. Overall, the results indicate that NGF activates the p75 receptor to cause a prolonged increase in Ca(2+) influx through L-type channels, which in turn up-regulates the prolactin mRNA. PMID- 16690704 TI - Developing a head for energy sensing: AMP-activated protein kinase as a multifunctional metabolic sensor in the brain. AB - The 5'-adenosine monophosphate-activated protein kinase (AMPK) is a metabolic and stress sensor that has been functionally conserved throughout eukaryotic evolution. Activation of the AMPK system by various physiological or pathological stimuli that deplete cellular energy levels promotes activation of energy restorative processes and inhibits energy consumptive processes. AMPK has a prominent role not only as a peripheral sensor of energy balance, but also in the CNS as a multifunctional metabolic sensor. Recent work suggests that AMPK plays an important role in maintaining whole body energy balance by coordinating feeding behaviour through the hypothalamus in conjunction with peripheral energy expenditure. In addition, brain AMPK is activated by energy-poor conditions induced by hypoxia, starvation, and ischaemic stroke. Under these conditions, AMPK is activated as a protective response in an attempt to restore cellular homeostasis. However in vivo, it appears that the overall consequence of activation of AMPK is more complex than previously imagined, in that over activation may be deleterious rather than neuroprotective. This review discusses recent findings that support the role of AMPK in brain as a multidimensional energy sensor and the consequences of its activation or inhibition under physiological and pathological states. PMID- 16690705 TI - Role of AMPK in skeletal muscle metabolic regulation and adaptation in relation to exercise. AB - The 5'-AMP-activated protein kinase (AMPK) is a potent regulator of skeletal muscle metabolism and gene expression. AMPK is activated both in response to in vivo exercise and ex vivo contraction. AMPK is therefore believed to be an important signalling molecule in regulating muscle metabolism during exercise as well as in adaptation of skeletal muscle to exercise training. The first part of this review is focused on different mechanisms regulating AMPK activity during muscle work such as alterations in nucleotide concentrations, availability of energy substrates and upstream AMPK kinases. We furthermore discuss the possible role of AMPK as a master switch in skeletal muscle metabolism with the main focus on AMPK in metabolic regulation during muscle work. Finally, AMPK has a well established role in regulating expression of genes encoding various enzymes in muscle, and this issue is discussed in relation to adaptation of skeletal muscle to exercise training. PMID- 16690706 TI - AMPK alterations in cardiac physiology and pathology: enemy or ally? AB - AMP-activated protein kinase (AMPK) has emerged as a key regulator of energy metabolism in the heart. The high energy demands of the heart are primarily met by the metabolism of both fatty acids and glucose, both processes being regulated by AMPK. During myocardial ischaemia a rapid activation of AMPK occurs, resulting in an activation of both glucose uptake and glycolysis, as well as an increase in fatty acid oxidation. This activation of AMPK has the potential to increase energy production and to inhibit apoptosis, thereby protecting the heart during the ischaemic stress. However, at clinically relevant high levels of fatty acids, ischaemic-induced activation of AMPK also stimulates fatty acid oxidation during and following ischaemia. This can contribute to ischaemic injury secondary to an inhibition of glucose oxidation, which results in a decrease in cardiac efficiency. In a number of other non-cardiac tissues, AMPK has been shown to have pro-apoptotic effects. As a result, the question of whether AMPK activation benefits or harms the ischaemic heart remains controversial. The role of AMPK in cardiac hypertrophy is also controversial. Activation of AMPK inhibits protein synthesis, and may be an adaptive response to pathological cardiac hypertrophy. However, none of mouse models of AMPK deficiency (excluding those that may involve the gamma2 subunit mutations) demonstrate increased cardiac mass, suggesting that AMPK is not essential for restriction of cardiac growth. In addition to the potential effects of AMPK on myofibrillar hypertrophy associated with pressure overload, there is also controversy with respect to the cardiac hypertrophy associated with the gamma2 subunit mutations. In the cardiac hypertrophy associated with glycogen overload, both activating and inactivating mutations of AMPK in mice are associated with a marked cardiac hypertrophy. This review will address the issue of whether AMPK activation acts as an enemy or ally to the ischaemic and hypertrophied heart. Resolving this issue has important implications as to whether therapeutic approaches to protect the ischaemic heart should be developed which either activate or inhibit AMPK. PMID- 16690707 TI - Open probability of the epithelial sodium channel is regulated by intracellular sodium. AB - The regulation of epithelial Na(+) channel (ENaC) activity by Na(+) was studied in Xenopus oocytes using two-electrode voltage clamp and patch-clamp recording techniques. Here we show that amiloride-sensitive Na(+) current (I(Na)) is downregulated when ENaC-expressing cells are exposed to high extracellular [Na(+)]. The reduction in macroscopic Na(+) current is accompanied by an increase in the concentration of intracellular Na(+) ([Na(+)](i)) and is only slowly reversible. At the single-channel level, incubating oocytes in high-Na(+) solution reduces open probability (P(o)) approximately twofold compared to when [Na(+)] is kept low, by increasing mean channel closed times. However, increasing P(o) by introducing a mutation in the beta-subunit (S518C) which, in the presence of [2-(trimethylammonium) ethyl] methane thiosulfonate (MTSET), locks the channel in an open state, could not alone abolish the downregulation of macroscopic current measured with exposure to high external [Na(+)]. Inhibition of the insertion of new channels into the plasma membrane using Brefeldin A revealed that surface channel lifetime is also markedly reduced under these conditions. In channels harbouring a beta-subunit mutation, R564X, associated with Liddle's syndrome, open probability in both high- and low-Na(+) conditions is significantly higher than in wild-type channels. Increasing the P(o) of these channels with an activating mutation abrogated the difference in macroscopic current observed between groups of oocytes incubated in high- and low-Na(+) conditions. These findings demonstrate that reduction of ENaC P(o) is a physiological mechanism limiting Na(+) entry when [Na(+)](i) is high. PMID- 16690708 TI - Short-term reorganization of input-deprived motor vibrissae representation following motor disconnection in adult rats. AB - It has been proposed that abnormal vibrissae input to the motor cortex (M1) mediates short-term cortical reorganization after facial nerve lesion. To test this hypothesis, we cut first the infraorbital nerve (ION cut) and then the facial nerve (VII cut) in order to evaluate M1 reorganization without any aberrant, facial-nerve-lesion-induced sensory feedback. In each animal, M1 output was assessed in both hemispheres by mapping movements induced by intracortical microstimulation. M1 output was compared in different types of peripheral manipulations: (i) contralateral intact vibrissal pad (intact hemispheres), (ii) contralateral VII cut (VII hemispheres), (iii) contralateral ION cut (ION hemispheres), (iv) contralateral VII cut after contralateral ION cut (ION + VII hemispheres), (v) contralateral pad botulinum-toxin-injected after ION cut (ION + BTX hemispheres). Right and left hemispheres in untouched animals were the reference for normal M1 map (control hemispheres). Findings demonstrated that: (1) in ION hemispheres, the mean size of the vibrissae representation was not significantly different from those in intact and control hemispheres; (2) reorganization of the vibrissae movement representation clearly emerged only in hemispheres where the contralateral vibrissae pad had undergone motor output disconnection (VII cut hemispheres); (3) the persistent loss of vibrissae input did not change the M1 reorganization pattern during the first 48 h after motor paralysis (ION + VII cut and ION + BTX hemispheres). Thus, after motor paralysis, vibrissa input does not provide the gating signal necessary to trigger M1 reorganization. PMID- 16690709 TI - Neurosteroid modulation of respiratory rhythm in rats during the perinatal period. AB - Neurosteroids regulate neuronal excitability and are expressed at particularly high levels in the CNS during the perinatal period. Further, neurosteroid levels are increased by a variety of stressors including hypoxia, asphyxia, parturition, ethanol exposure and infection. One mechanism by which neurosteroids regulate neuronal activity is by negative or positive modulation of GABA(A) receptor function. Perinatal respiration is strongly modulated by GABAergic synaptic drive, and GABA release is increased during hypoxia to contribute to hypoxia induced depression of neonatal ventilation. Here, we use in vitro and in vivo rat models to test the hypothesis that GABA(A) receptor-mediated modulation of perinatal respiration is markedly influenced by the presence of neurosteroids. The principal finding of this study was that the efficacy of GABA(A) receptor mediated modulation of respiratory membrane potential and rhythmogenesis is markedly enhanced by allopregnanolone and depressed by dehydroepiandrosterone sulphate. These data demonstrate that the modulation of breathing via GABA(A) receptor activation will be determined by the overall balance of negative and positive neurosteroid modulators within respiratory nuclei. This adds a level of complexity that must be considered when examining the depression of breathing in mammals associated with various behavioural states and pathogenic conditions such as apnoea and sudden death suspected to be associated with central respiratory dysfunction. PMID- 16690710 TI - The role of adenosine in the early respiratory and cardiovascular changes evoked by chronic hypoxia in the rat. AB - Experiments were performed on anaesthetized normoxic (N) rats and chronically hypoxic rats that had been exposed to 12% O2 for 1, 3 or 7 days (1, 3 or 7CH rats). The adenosine A1 receptor antagonist DPCPX did not affect the resting hyperventilation of 1-7CH rats breathing 12% O2 and increased resting heart rate (HR) in 1CH rats only. DPCPX partially restored the decreased baseline arterial pressure (ABP) and increased femoral vascular conductance (FVC) of 1 and 3CH rats, but had no effect in N or 7CH rats. DPCPX also attenuated the decrease in arterial blood pressure (ABP) and increase in FVC evoked by acute hypoxia in N and 1-7CH rats. The non-selective adenosine receptor antagonist 8-SPT had no further effect on baselines or cardiovascular responses to acute hypoxia, but attenuated the hypoxia-evoked increase in respiratory frequency in 1-7CH rats. In N, and 1 and 3CH rats, the inducible nitric oxide synthase (iNOS) inhibitor aminoguanidine had no effect on baselines or increases in FVC evoked by acetylcholine. We propose: (i) that tonically released adenosine acting on A1 receptors reduces HR in 1CH rats and stimulates endothelial NOS in 1 and 3CH rats to decrease ABP and increase FVC, the remaining NO-dependent tonic vasodilatation being independent of iNOS activity; (ii) that in 7CH rats, tonic adenosine release has waned; (iii) that in 1-7CH rats, adenosine released by acute hypoxia stimulates A1 but not A2 receptors to produce muscle vasodilatation, and stimulates carotid body A2 receptors to increase respiration. PMID- 16690711 TI - The early effects of chronic hypoxia on the cardiovascular system in the rat: role of nitric oxide. AB - Experiments were performed under Saffan anaesthesia on normoxic (N) rats and on chronically hypoxic rats exposed to 12% O2 for 1, 3 or 7 days (1, 3 or 7CH rats): N rats routinely breathed 21% O2 and CH rats 12% O2. The 1, 3 and 7CH rats showed resting hyperventilation relative to N rats, but baseline heart rate (HR) was unchanged and arterial blood pressure (ABP) was lowered. Femoral vascular conductance (FVC) was increased in 1 and 3CH rats, but not 7CH rats. When 1-7CH rats were acutely switched to breathing 21% O2 for 5 min, ABP increased and FVC decreased, consistent with removal of a hypoxic dilator stimulus that is waning in 7CH rats. We propose that this is because the increase in haematocrit and vascular remodelling in skeletal muscle help restore the O2 supply. The increases in FVC evoked by acute hypoxia (8% O2 for 5 min) and by infusion for 5 min of alpha-calcitonin gene-related peptide (alpha-CGRP), which are NO-dependent, were particularly accentuated in 1CH, relative to N rats. The NO synthesis inhibitor L NAME increased ABP, decreased HR and greatly reduced FVC, and attenuated increases in FVC evoked by acute hypoxia and alpha-CGRP, such that baselines and responses were similar in N and 1-7CH rats. We propose that in the first few days of chronic hypoxia there is tonic NO-dependent vasodilatation in skeletal muscle that is associated with accentuated dilator responsiveness to acute hypoxia and dilator substances that are NO -dependent. PMID- 16690712 TI - Glutamatergic synapses in the rat nucleus tractus solitarii develop by direct insertion of calcium-impermeable AMPA receptors and without activation of NMDA receptors. AB - Calcium influxes through ionotropic glutamate receptors (AMPA and NMDA receptors, AMPARs and NMDARs) are considered to be critical for the shaping and refinement of neural circuits during synaptogenesis. Using a combined morphological and electrophysiological approach, we evaluated this hypothesis at the level of the nucleus tractus solitarii (NTS), a brainstem structure that is a gateway for many visceral sensory afferent fibres. We confirmed that in the NTS, the first excitatory synapses appeared at embryonic day 18. We next characterized the biophysical properties of NTS AMPARs. Throughout perinatal development, both evoked and miniature EPSCs recorded in the presence of an NMDAR blocker were insensitive to polyamines and had linear current-voltage relationships. This demonstrated that AMPARs at NTS excitatory synapses were calcium-impermeable receptors composed of a majority of GluR2 subunits. We then investigated the influence of calcium influxes through NMDARs on the development of NTS synaptic transmission. We found that NMDAR expression at synaptic sites did not precede AMPAR expression. Moreover, NMDAR blockade in utero did not prevent the development of AMPAR synaptic currents and the synaptic clustering of GluR2 subunits. Thus, our data support an alternative model of synaptogenesis that does not depend on calcium influxes through either AMPARs or NMDARs. This model may be particularly relevant to the formation of neural networks devoted to basic behaviours required at birth for survival. PMID- 16690713 TI - Effects of repetitive TMS on visually evoked potentials and EEG in the anaesthetized cat: dependence on stimulus frequency and train duration. AB - Repetitive transcranial magnetic stimulation (rTMS) has been shown to alter cortical excitability that lasts beyond the duration of rTMS application itself. High-frequency rTMS leads primarily to facilitation, whereas low-frequency rTMS leads to inhibition of the treated cortex. However, the contribution of rTMS train duration is less clear. In this study, we investigated the effects of nine different rTMS protocols, including low and high frequencies, as well as short and long applications (1, 3 and 10 Hz applied for 1, 5 and 20 min), on visual cortex excitability in anaesthetized and paralysed cats by means of visual evoked potential (VEP) and electroencephalography (EEG) recordings. Our results show that 10 Hz rTMS applied for 1 and 5 min significantly enhanced early VEP amplitudes, while 1 and 3 Hz rTMS applied for 5 and 20 min significantly reduced them. No significant changes were found after 1 and 3 Hz rTMS applied for only 1 min, and 10 Hz rTMS applied for 20 min. EEG activity was only transiently (<20 s) affected, with increased delta activity after 1 and 3 Hz rTMS applied for 1 or 5 min. These findings indicate that the effects of rTMS on cortical excitability depend on the combination of stimulus frequency and duration (or total number of stimuli): short high-frequency trains seem to be more effective than longer trains, and low-frequency rTMS requires longer applications. Changes in the spectral composition of the EEG were not correlated to changes in VEP size. PMID- 16690714 TI - The contribution of brain angiotensin II to the baroreflex regulation of renal sympathetic nerve activity in conscious normotensive and hypertensive rats. AB - Angiotensin II receptor density in the brain is elevated when dietary salt intake is raised or in the state of hypertension. The aim of this study was to evaluate whether the angiotensin II modulation of the baroreceptor control of renal sympathetic nerve activity was altered under these conditions. Wistar rats, fed either a regular (0.25% w/w sodium) or high-salt diet (3.1% w/w sodium), or stroke-prone spontaneously hypertensive rats (SHRSPs) were implanted with cannulae in the carotid artery, jugular vein and the cerebroventricle and with recording electrodes on the renal sympathetic nerves. Three days later, baroreceptor gain curves were generated for renal sympathetic nerve activity and heart rate before and following intracerebroventricular (i.c.v.) administration of losartan (15 mug) to block angiotensin AT1 receptors. The rats fed a regular diet had a mean blood pressure of 116 +/- 3 mmHg and heart rate of 467 +/- 25 beats min(-1), which remained unchanged after the i.c.v. administration of losartan. The sensitivity or curvature coefficient of the baroreceptor curve for renal sympathetic nerve activity was increased by 36% (P < 0.05) following losartan. In the rats fed a high-salt diet, all cardiovascular variables and the losartan-induced increase in the baroreceptor curvature coefficient for renal sympathetic nerve activity (29%) were similar to values in rats on the regular sodium diet. The heart rate baroreceptor curvature coefficient was not altered in either the rats fed a regular or a high-salt diet. The slope of the renal sympathetic nerve activity baroreflex gain curve in the SHRSPs was less and the increase following administration of losartan (54%) was greater than in the Wistar rats. These data indicate that in the conscious state, the tonic inhibitory action of brain angiotensin II on the baroreflex regulation of renal sympathetic nerve activity was unaffected by raised dietary sodium, but its role was enhanced in the SHRSPs. PMID- 16690715 TI - Excitation of rat hippocampal interneurons via modulation of endogenous agonist activity at the alpha7 nicotinic ACh receptor. AB - The alpha7 subtype of the nicotinic acetylcholine receptor (alpha7 nAChR) is prominently expressed in the hippocampus where it is thought to play a role in the regulation of cognitive function. In this study, we have investigated the effects of 5-hydroxyindole (5-HI), a positive modulator of the alpha7 nAChR, on GABAergic activity in hippocampal CA1 stratum radiatum interneurons in acute rat brain slices. Superfusion of 5-HI (100 microM) increased the mean frequency and amplitude of spontaneous IPSCs (sIPSCs). The potentiation was occluded by pretreatment of slices with: (1) a high concentration of the broad-spectrum agonist nicotine to desensitize the alpha7 receptor, (2) an alpha7 nAChR antagonist, and (3) tetrodotoxin to block action potential firing. These results indicate that facilitation by 5-HI was mediated by the alpha7 nAChR and required neuronal excitation. In contrast, 5-HI had no effect on sIPSCs recorded in hippocampal slices from younger animals, even though the expression of functional alpha7 nAChRs was confirmed by agonist application experiments. In these slices, 5-HI only enhanced sIPSCs after pretreatment with the acetylcholinesterase inhibitor Bw284c51. Taken together, our results suggest that 5-HI facilitates GABAergic transmission via excitation of the alpha7 nAChR, and that this effect requires the presence of the endogenous agonist ACh in the extracellular environment of the receptor. PMID- 16690716 TI - Hyperpolarization-activated cation channels in fast-spiking interneurons of rat hippocampus. AB - Hyperpolarization-activated channels (Ih or HCN channels) are widely expressed in principal neurons in the central nervous system. However, Ih in inhibitory GABAergic interneurons is less well characterized. We examined the functional properties of Ih in fast-spiking basket cells (BCs) of the dentate gyrus, using hippocampal slices from 17- to 21-day-old rats. Bath application of the Ih channel blocker ZD 7288 at a concentration of 30 microm induced a hyperpolarization of 5.7 +/- 1.5 mV, an increase in input resistance and a correlated increase in apparent membrane time constant. ZD 7288 blocked a hyperpolarization-activated current in a concentration-dependent manner (IC50, 1.4 microm). The effects of ZD 7288 were mimicked by external Cs+. The reversal potential of Ih was -27.4 mV, corresponding to a Na+ to K+ permeability ratio (PNa/PK) of 0.36. The midpoint potential of the activation curve of Ih was -83.9 mV, and the activation time constant at -120 mV was 190 ms. Single-cell expression analysis using reverse transcription followed by quantitative polymerase chain reaction revealed that BCs coexpress HCN1 and HCN2 subunit mRNA, suggesting the formation of heteromeric HCN1/2 channels. ZD 7288 increased the current threshold for evoking antidromic action potentials by extracellular stimulation, consistent with the expression of Ih in BC axons. Finally, ZD 7288 decreased the frequency of miniature inhibitory postsynaptic currents (mIPSCs) in hippocampal granule cells, the main target cells of BCs, to 70 +/- 4% of the control value. In contrast, the amplitude of mIPSCs was unchanged, consistent with the presence of Ih in inhibitory terminals. In conclusion, our results suggest that Ih channels are expressed in the somatodendritic region, axon and presynaptic elements of fast-spiking BCs in the hippocampus. PMID- 16690717 TI - Influence of breathing route on upper airway lining liquid surface tension in humans. AB - We have recently demonstrated that the severity of sleep-disordered breathing in obstructive sleep apnoea hypopnoea syndrome (OSAHS) can be reduced by lowering the surface tension (gamma) of the upper airway lining liquid (UAL). Morning xerostomia (related to oral breathing during sleep) is reported by most OSAHS patients. In the present study we examine relationships between breathing route, oral mucosal 'wetness' and the gamma of UAL. We studied eight healthy subjects (age, 25 +/- 5 years [mean +/- S.D.]; body-mass index, 23 +/- 2 kg m(-2)) during a 120 min challenge of both nasal-only breathing (mouth taped) and oral-only breathing (nose clip), each on a separate day (randomized). Both oral mucosal 'wetness' (5 s contact gravimetric absorbent paper strip method) and the gamma ('pull-off' force technique) of 0.2 microl samples of UAL obtained from the posterior pharyngeal wall were measured at 15 min intervals (mouth tape removed and replaced as required). Upper airway mucosal 'wetness' increased during 120 min of nasal breathing from 4.0 +/- 0.4 (mean +/- S.E.M.) to 5.3 +/- 0.3 microl (5 s)(-1) but decreased from 4.5 +/- 0.4 to 0.1 +/- 0.2 microl (5 s)(-1) with oral breathing (both P < 0.001, repeated-measures ANOVA, Tukey's multiple comparison test, post hoc test). Concurrently, the gamma of UAL decreased from 59.3 +/- 2.2 to 51.8 +/- 0.98 mN m(-1) with nasal breathing but increased from 64.4 +/- 2.7 to 77.4 +/- 1.1 mN m(-1) with oral breathing (P < 0.001). For the group and all conditions studied, gamma of UAL values strongly correlated with upper airway mucosal 'wetness' (correlation coefficient, r2 = -0.34, P < 0.001; linear regression). We conclude that oral breathing increases and nasal breathing decreases the gamma of UAL in healthy subjects during wakefulness. We speculate that nasal breathing in OSAHS patients during sleep may promote a low gamma of UAL that may contribute to reducing the severity of sleep-disordered breathing. PMID- 16690718 TI - Identification of novel small molecule inhibitors of amyloid precursor protein synthesis as a route to lower Alzheimer's disease amyloid-beta peptide. AB - A wealth of independent research with transgenic mice, antibodies, and vaccines has pointed to a causative role of the amyloid-beta peptide (A beta) in Alzheimer's disease (AD). Based on these and earlier associative studies, A beta represents a promising target for development of therapeutics focused on AD disease progression. Interestingly, a cholinesterase inhibitor currently in clinical trials, phenserine, has been shown to inhibit production of both amyloid precursor protein (APP) and A beta. We have shown that this inhibition occurs at the post-transcriptional level with a specific blocking of the synthesis of APP relative to total protein synthesis (Shaw et al., 2001). However, the dose of phenserine necessary to block APP production is far higher than that needed to elicit its anticholinesterase activity, and it is these latter actions that are dose limiting in vivo. The focus of this study was to screen 144 analogs of phenserine to identify additional small molecules that inhibit APP protein synthesis, and thereby A beta production, without possessing potent acetylcholinesterase (AChE) inhibitory activity. An enzyme-linked immunosorbent assay was used to identify analogs capable of suppressing APP production following treatment of human neuroblastoma cells with 20 muM of compound. Eight analogs were capable of dose dependently reducing APP and A beta production without causing cell toxicity in further studies. Several of these analogs had little to no AChE activities. Translation of APP and A beta actions to mice was demonstrated with one agent. They thus represent interesting lead molecules for assessment in animal models, to define their tolerance and utility as potential AD therapeutics. PMID- 16690719 TI - Megalin-dependent internalization of cadmium-metallothionein and cytotoxicity in cultured renal proximal tubule cells. AB - Chronic cadmium (Cd2+) exposure results in renal proximal tubular cell damage. Delivery of Cd2+ to the kidney occurs mainly as complexes with metallothionein-1 (molecular mass approximately 7 kDa), freely filtered at the glomerulus. For Cd2+ to gain access to the proximal tubule cells, these complexes are thought to be internalized via receptors for small protein ligands, such as megalin and cubilin, followed by release of Cd2+ from metallothionein-1 in endosomal/lysosomal compartments. To investigate the role of megalin in renal cadmium-metallothionein-1 reabsorption, megalin expression and dependence of cadmium-metallothionein-1 internalization and cytotoxicity on megalin were studied in a renal proximal tubular cell model (WKPT-0293 Cl.2 cells). Expression of megalin was detected by reverse transcriptase-polymerase chain reaction and visualized by immunofluorescence both at the cell surface (live staining) and intracellularly (permeabilized cells). Internalization of Alexa Fluor 488-coupled metallothionein-1 was concentration-dependent, saturating at approximately 15 microM. At 14.3 microM, metallothionein-1 uptake could be significantly attenuated by 30.9 +/- 6.6% (n = 4) by 1 muM of the receptor-associated protein (RAP) used as a competitive inhibitor of cadmium-metallothionein-1 binding to megalin and cubilin. Consistently, cytotoxicity of a 24-h treatment with 7.14 muM cadmium-metallothionein-1 was significantly reduced by 41.0 +/- 7.6%, 61.6 +/- 3.4%, and 26.2 +/- 1.8% (n = 4-5 each) by the presence of 1 microM RAP, 400 microg/ml anti-megalin antibody, or 5 microM of the cubilin-specific ligand, apo transferrin, respectively. Cubilin expression in proximal tubule cells was also confirmed at the mRNA and protein level. The data indicate that renal proximal tubular cadmium-metallothionein-1 uptake and cell death are mediated at least in part by megalin. PMID- 16690720 TI - Mu-delta opioid receptor functional interaction: Insight using receptor-G protein fusions. AB - Fusion proteins between a receptor and a pertussis toxin-insensitive G(i)alpha subunit were used to gain insight into the molecular interactions that take place upon mu and delta opioid receptor heterodimerization. When mu opioid receptor G(i1)alpha fusions were coexpressed with nonfused delta opioid receptors in human embryonic kidney 293 cells, or vice versa, receptor heterodimers were detected by coimmunoprecipitation. In pertussis toxin-treated cells, receptor coexpression decreased the amount of guanosine 5'-O-(3-[35S]thio)triphosphate ([35S]GTPgammaS) incorporated in the fused G alpha protein after the addition of agonists specific for the receptor-G(i1)alpha fusion. In addition, activation of the G alpha protein occurred in heterodimers upon addition of an agonist specific for the nonfused receptor. It remained unaffected by an inverse agonist specific for the receptor-G(i1)alpha fusion. These data suggest that signaling through the receptor-G(i1)alpha fusion protein is impaired in heterodimers and support a mechanism in which activation of the G alpha subunit is promoted by a direct interaction with the nonfused receptor. Alternatively, receptor coexpression did not modify the ligand binding properties for the high-affinity state of the receptor-G(i1)alpha fusion nor the EC50 values for agonist-induced [35S]GTPgammaS incorporation in the G(i1)alpha subunit. In addition, no binding competition was observed between delta and mu ligands. Together, the data point to mu-delta opioid receptor heterodimers formed by contact interactions between monomers that retain their structural integrity. PMID- 16690721 TI - Endoxifen, a secondary metabolite of tamoxifen, and 4-OH-tamoxifen induce similar changes in global gene expression patterns in MCF-7 breast cancer cells. AB - We recently demonstrated that endoxifen (4-hydroxy-N-desmethyl-tamoxifen), a pharmacogenetically regulated metabolite of tamoxifen, is equipotent to 4-hydroxy tamoxifen (4-OH-Tam) with respect to estrogen receptor binding and inhibition of 17beta-estradiol (E2)-induced cell proliferation. Endoxifen was also found to be more abundant in human plasma than 4-OH-Tam, and its formation has been shown to be primarily catalyzed by cytochrome P450 2D6 (CYP2D6). Here, we report studies evaluating the effects of endoxifen, 4-OH-Tam, and E2 on gene expression in MCF-7 cells using Affymetrix U133A GeneChip Arrays (Santa Clara, CA). We detected 4062 genes that were E2-regulated (1924 induced; 2138 suppressed), and the ratio of E2 induced versus E2-suppressed genes was consistent regardless of the cutoff value. In the presence of E2, 2444 and 2390 genes were affected by 4-OH-Tam and endoxifen, respectively, when no minimal -fold change cutoff was implemented. The majority of genes regulated by the tamoxifen metabolites were also E2-responsive (74.4 and 73.3%, respectively). Endoxifen and 4-OH-Tam had overlapping effects on 1365 E2-sensitive genes, whose -fold effects between these metabolites were highly correlated (R2 = 0.99). A significant correlation was also found between the -fold effects of 249 E2-insensitive genes coregulated by both metabolites (R2 = 0.99). Hierarchical clustering analysis demonstrated similar gene regulation patterns between these metabolites, which were distinct from E2 or vehicle treatment patterns. Using real time-polymerase chain reaction, we validated the gene expression patterns of five genes that were differentially regulated by endoxifen and 4-OH-Tam. We conclude that endoxifen and 4-OH-Tam have similar effects on global gene expression patterns in MCF-7 cells and that the majority of the affected genes are estrogen-regulated genes. PMID- 16690722 TI - Role of matrix metalloproteinases in the inflammatory response in human airway cell-based assays and in rodent models of airway disease. AB - Since the discovery of the first matrix metalloproteinase (MMP), this ever growing family of proteinases has been the subject of intense research. Although it was initially believed that MMPs were solely involved in matrix turnover and degradation, there are now data suggesting MMPs are actively involved in the inflammatory process. In previous studies, we have demonstrated an increase in MMP expression in human cell-based assays and in preclinical rat models of airway inflammation. Therefore, the aim of this study was to characterize the role of MMPs in these models by profiling the impact of a broad-spectrum MMP inhibitor. In lipopolysaccharide (LPS)-stimulated THP-1 cells and primary human lung tissue macrophages, the MMP inhibitor had no significant effect on the release of tumor necrosis factor-alpha, interleukin (IL)-8, IL-1 beta, growth-regulated oncogene alpha, macrophage inflammatory protein-1 alpha, or IL-6 whereas dexamethasone has a significant impact on all cytokines from both cell types. Similarly, in the more biologically complex LPS-driven rat model of airway inflammation, the MMP inhibitor did not have an impact on mediator release and cellular burden. The compound did, however, significantly reduce levels of lung MMP-9. Furthermore, in a "disease" model, the compound did not affect cellular inflammation but did significantly reduce elastase-induced experimental emphysema. In summary, these data demonstrate for the first time that MMPs do not play a role in the increase in inflammatory mediators or cellular burden observed in these preclinical models. However, they do appear to be involved in the elastase-driven breakdown of airway structure, which is not due to a direct effect of the stimulus. PMID- 16690723 TI - Effect of vagus nerve stimulation on serotonergic and noradrenergic transmission. AB - Vagus nerve stimulation (VNS) is an antiepileptic treatment, which has recently shown promise as an antidepressant. Yet, its antidepressant mechanisms of action are unknown. Serotonergic [5-hydroxytryptamine (5-HT, serotonin)] and noradrenergic [norepinephrine (NE)] systems are involved in the pathophysiology of depression and in the mechanisms of action of antidepressants. The present study analyzes 5-HT and NE neuronal firing rates in their brainstem nuclei: the dorsal raphe nucleus (DRN) and locus coeruleus (LC), respectively. The basal firing rates in the DRN and LC were significantly increased after long-term treatments with VNS. After short-term VNS treatments, firing rates were significantly higher for LC (at 1 h and 3 days). As changes in their firing rate may have been due to altered autoreceptor sensitivities, the responses of autoreceptors to the acute administration of their respective agonists were assessed. However, no significant difference was seen in the DRN. No significant differences in dose response curves for 5-HT(1A) somatodendritic and alpha 2 adrenergic autoreceptors were noticed between long-term VNS and controls. VNS appears to have a novel mechanism of antidepressant action, enabling its effectiveness in treatment-resistant depression. LC firing rates significantly increase earlier than the DRN basal firing. As the LC has an excitatory influence on DRN, it is possible that the increased DRN firing rate is secondary to an initial increased LC firing rate from VNS. PMID- 16690724 TI - Hepatobiliary disposition of a drug/metabolite pair: Comprehensive pharmacokinetic modeling in sandwich-cultured rat hepatocytes. AB - The hepatobiliary disposition of xenobiotics may involve passive and/or active uptake, metabolism by cytochromes P450, and excretion of the parent compound and/or metabolite(s) into bile. Although in vitro systems have been used to evaluate these individual processes discretely, mechanistic in vitro studies of the sequential processes of uptake, metabolism, and biliary or basolateral excretion are limited. The current studies used sandwich-cultured (SC) rat hepatocytes combined with a comprehensive pharmacokinetic modeling approach to investigate the hepatobiliary disposition of terfenadine and fexofenadine, a model drug/metabolite pair. The metabolism of terfenadine and the biliary excretion of terfenadine and fexofenadine were determined in control and dexamethasone-treated SC rat hepatocytes. Dexamethasone (DEX) treatment increased the formation rates of the terfenadine metabolites azacyclonol and fexofenadine approximately 20- and 2-fold, respectively. The biliary excretion index (BEI) of fexofenadine, when generated by terfenadine metabolism, was not significantly different from the BEI of preformed fexofenadine (15 +/- 2% versus 19 +/- 2%, respectively). Pharmacokinetic modeling revealed that the rate constant for hepatocyte uptake was faster for terfenadine compared with preformed fexofenadine (2.5 versus 0.08 h(-1), respectively), whereas the biliary excretion rate constant for preformed fexofenadine exceeded that of terfenadine (0.44 versus 0.039 h(-1), respectively). Interestingly, the rate constants for basolateral excretion of terfenadine and fexofenadine were comparable (3.2 versus 1.9 h(-1), respectively) and increased only slightly with DEX treatment. These studies demonstrate the utility of the SC hepatocyte model, coupled with pharmacokinetic modeling, to evaluate the hepatobiliary disposition of generated metabolites. PMID- 16690725 TI - Pharmacological characterization of the competitive GLUK5 receptor antagonist decahydroisoquinoline LY466195 in vitro and in vivo. AB - The excitatory neurotransmitter glutamate has been implicated in both migraine and persistent pain. The identification of the kainate receptor GLU(K5) in dorsal root ganglia, the dorsal horn, and trigeminal ganglia makes it a target of interest for these indications. We examined the in vitro and in vivo pharmacology of the competitive GLU(K5)-selective kainate receptor antagonist LY466195 [(3S,4aR,6S,8aR)-6-[[(2S)-2-carboxy-4,4-difluoro-1-pyrrolidinyl]-methyl]decahydro 3-isoquinolinecarboxylic acid)], the most potent GLU(K5) antagonist described to date. Comparisons were made to the competitive GLU(K5)/alpha-amino-3-hydroxy-5 methyl-4-isoxazole propionic acid (AMPA) receptor antagonist LY293558 [(3S,4aR,6R,8aR)-6-[2-(1(2)H-tetrazole-5-yl)ethyl]-decahydroisoquinoline-3 carboxylic acid], other decahydroisoquinoline GLU(K5) receptor antagonists, and the noncompetitive AMPA receptor antagonist LY300168 [1-(4-aminophenyl)-4-methyl 7,8-methylenedioxy-5H-2,3-benzodi-azepine]. When characterized electrophysiologically in rat dorsal root ganglion neurons, LY466195 antagonized kainate (30 microM)-induced currents with an IC50 value of 0.045 +/- 0.011 microM. In HEK293 cells transfected with GLU(K5), GLU(K2)/GLU(K5), or GLU(K5)/GLU(K6) receptors, LY466195 produced IC50 values of 0.08 +/- 0.02, 0.34 +/- 0.17, and 0.07 +/- 0.02 microM, respectively. LY466195 was efficacious in a dural plasma protein extravasation (PPE) model of migraine with an ID100 value of 100 microg/kg i.v. LY466195 was also efficacious in the c-fos migraine model, with a dose of 1 microg/kg i.v. significantly reducing the number of Fos-positive cells in the rat nucleus caudalis after electrical stimulation of the trigeminal ganglion. Furthermore, LY466195 showed no contractile activity in the rabbit saphenous vein in vitro. The diethyl ester prodrug of LY466195 was also efficacious in the same PPE and c-fos models after oral administration at doses of 10 and 100 microg/kg, respectively while having no N-methyl-D-aspartate antagonist-like behavioral effects at oral doses up to 100 mg/kg. PMID- 16690726 TI - Hereditary haemorrhagic telangiectasia: mutation detection, test sensitivity and novel mutations. AB - BACKGROUND: Hereditary haemorrhagic telangiectasia (HHT) is a genetic disorder present in 1 in 8000 people and associated with arteriovenous malformations. Genetic testing can identify individuals at risk of developing the disease and is a useful diagnostic tool. OBJECTIVE: To present a strategy for mutation detection in families clinically diagnosed with HHT. METHODS: An optimised strategy for detecting mutations that predispose to HHT is presented. The strategy includes quantitative multiplex polymerase chain reaction, sequence analysis, RNA analysis, validation of missense mutations by amino acid conservation analysis for the ENG (endoglin) and ACVRL1 (ALK1) genes, and analysis of an ACVRL1 protein structural model. If no causative ENG or ACVRL1 mutation is found, proband samples are referred for sequence analysis of MADH4 (associated with a combined syndrome of juvenile polyposis and HHT). RESULTS: Data obtained over the past eight years were summarised and 16 novel mutations described. Mutations were identified in 155 of 194 families with a confirmed clinical diagnosis (80% sensitivity). Of 155 mutations identified, 94 were in ENG (61%), 58 in ACVRL1 (37%), and three in MADH4 (2%). CONCLUSIONS: For most missense variants of ENG and ACVRL1 reported to date, study of amino acid conservation showed good concordance between prediction of altered protein function and disease occurrence. The 39 families (20%) yet to be resolved may carry ENG, ACVRL1, or MADH4 mutations too complex or difficult to detect, or mutations in genes yet to be identified. PMID- 16690727 TI - Very mild cases of Rett syndrome with skewed X inactivation. AB - BACKGROUND: Rett syndrome, a common cause of mental retardation in females, is caused by mutations in the MECP2 gene. Most females with MECP2 mutations fulfil the established clinical criteria for Rett syndrome, but single cases of asymptomatic carriers have been described. It is therefore likely that there are individuals falling between these two extreme phenotypes. OBJECTIVE: To describe three patients showing only minor symptoms of Rett syndrome. FINDINGS: The patient with the best intellectual ability had predominantly psychiatric problems with episodes of uncontrolled aggression that have not been described previously in individuals with MECP2 mutations. All three patients had normal hand function, communicated well, and showed short spells of hyperventilation only under stress. Diagnosis in such individuals requires the identification of subtle signs of Rett syndrome in girls with a mild mental handicap. Analysis of the MECP2 gene revealed mutations that are often found in classical Rett syndrome. Skewed X inactivation was present in all three cases, which may explain the mild phenotype. CONCLUSIONS: Because of skewed X inactivation, the phenotype of Rett patients may be very mild and hardly recognisable. PMID- 16690729 TI - Prenatal diagnosis of myopathy, encephalopathy, lactic acidosis, and stroke-like syndrome: contribution to understanding mitochondrial DNA segregation during human embryofetal development. AB - INTRODUCTION: Myopathy, encephalopathy, lactic acidosis, and stroke-like (MELAS) syndrome, a maternally inherited disorder that is among the most common mitochondrial DNA (mtDNA) diseases, is usually associated with the m.3242A>G mutation of the mitochondrial tRNA(leu) gene. Very few data are available with respect to prenatal diagnosis of this serious disease. The rate of mutant versus wild-type mtDNA (heteroplasmy) in fetal DNA is indeed considered to be a poor indicator of postnatal outcome. MATERIALS AND METHODS: Taking advantage of a novel semi-quantitative polymerase chain reaction test for m.3243A>G mutant load assessment, we carried out nine prenatal diagnoses in five unrelated women, using two different fetal tissues (chorionic villi v amniocytes) sampled at two or three different stages of pregnancy. RESULTS: Two of the five women, although not carrying m.3243A>G in blood or extra-blood tissues, were, however, considered at risk for transmission of the mutation, as they were closely related to MELAS affected individuals. The absence of 3243A>G in the blood of first degree relatives was associated with no mutated mtDNA in the cardiovascular system (CVS) or amniocytes, and their three children are healthy, with a follow-up of 3 months 3 years. Among the six fetuses from the three carrier women, three were shown to be homoplasmic (0% mutant load), the remaining three being heteroplasmic, with a mutant load ranging from 23% to 63%. The fetal mutant load was fairly stable at two or three different stages of pregnancy in CVS and amniocytes. Although pregnancy was terminated in the case of the fetus with a 63% mutant load, all other children are healthy with a follow-up of 3 months-6 years. CONCLUSION: These data suggest that a prenatal diagnosis for MELAS syndrome might be helpful for at-risk families. PMID- 16690728 TI - Syndromes and constitutional chromosomal abnormalities associated with Wilms tumour. AB - Wilms tumour has been reported in association with over 50 different clinical conditions and several abnormal constitutional karyotypes. Conclusive evidence of an increased risk of Wilms tumour exists for only a minority of these conditions, including WT1 associated syndromes, familial Wilms tumour, and certain overgrowth conditions such as Beckwith-Wiedemann syndrome. In many reported conditions the rare co-occurrence of Wilms tumour is probably due to chance. However, for several conditions the available evidence cannot either confirm or exclude an increased risk, usually because of the rarity of the syndrome. In addition, emerging evidence suggests that an increased risk of Wilms tumour occurs only in a subset of individuals for some syndromes. The complex clinical and molecular heterogeneity of disorders associated with Wilms tumour, together with the apparent absence of functional links between most of the known predisposition genes, suggests that abrogation of a variety of pathways can promote Wilms tumorigenesis. PMID- 16690730 TI - Different human papillomavirus 16/18 infection in Chinese non-small cell lung cancer patients living in Wuhan, China. AB - BACKGROUND: Inconsistency in the prevalence of infection by human papillomavirus (HPV) in lung cancer patients was found between different countries with racial and geographic variations. Our previous reports have indicated that a high-risk HPV 16/18 DNA was frequently detected in Chinese lung cancer patients living in Taichung, Taiwan (Cheng et al. Cancer Res. 2001;61:2799-803). Thus, we conducted this study to verify whether there was a similar HPV 16/18 infection prevalence in lung cancer patients from Wuhan, China. METHODS: To reduce the false positive HPV detection, the paraffin sections of 73 lung tumors and 34 non-cancer controls from Wuhan, China were collected for detection of the presence of HPV 16/18 DNA by in situ hybridization (ISH). RESULTS: Our results showed that the rates of HPV 16 and/or 18 infections in patients with lung tumors were significantly higher than in 34 non-cancer control subjects (26.0 versus 2.8% for HPV 16, P = 0.030; 23.3 versus 5.7% for HPV 18, P = 0.031; 27.7 versus 5.9% for HPV 16 or 18, P = 0.003) with a similar infection frequency of HPV 16 and 18 types in lung tumors. This result indicated that HPV 16/18 infection may be associated with lung cancer development in Chinese patients from Wuhan, China. Further statistical analyses revealed that HPV 16 or 18 infection was not correlated with any clinico pathological parameter studied, including age, gender, smoking status, tumor type, tumor stage and tumor grades. Interestingly, smoking and male patients had a higher prevalence of HPV 16, although not reaching a statistical significance, compared with non-smoking and female patients, respectively (33.3% for smokers versus 20.0% non-smokers; 33.3% for male versus 17.6% for female). As compared with the HPV 16/18 infection in Taiwan, Chinese patients with lung cancer from Wuhan had a different HPV 16/18 infection prevalence. CONCLUSION: Difference in HPV 16/18 infection in lung cancer patients from Wuhan, China and Taichung, Taiwan suggests that HPV 16/18 might play a different role in lung cancer development among Chinese living in different areas. PMID- 16690731 TI - Genotyping of ovine prion protein gene (PRNP) variants by PCR with melting curve analysis. AB - BACKGROUND: Scrapie is the transmissible spongiform encephalopathy in sheep. Because genetic variants of the ovine PrP gene (PRNP) can be associated with disease risk, the European Union initiated programs to eradicate high-risk PRNP genotypes from sheep livestock. For this purpose, reliable and cost-effective genotyping is needed. METHODS: We amplified DNA to cover the 3 risk codons in exon 3 encoding amino acids 136, 154, and 171. Amplicons were mixed with dye labeled probe sets, and melting curves were recorded in a LightCycler by use of color and temperature multiplexing. Probe design was based on thermodynamic calculations to ensure unequivocal results for the 3 codons of interest, taking the additional F141 and T137 sequence variants into account. RESULTS: The fluorescence resonance energy transfer (FRET) method, when compared with sequencing, gave exactly the predicted melting temperatures for all possible genotypes. When we validated the method with samples from official certification programs, it showed completely matching results. Turnaround time was approximately 5 h after receipt of a whole-blood sample. The method detected the rare sequence variants T137 and F141, which were clearly distinguishable from the other known genotypes by melting curve analysis. One scrapie sheep was ARR/ARR, which is considered the haplotype with the lowest risk. CONCLUSIONS: The FRET based PRNP genotyping method for sheep is rapid and can differentiate all genotypes at each locus in 1 capillary. The assay is fast and has lower costs than restriction fragment length polymorphism analysis or sequencing. PMID- 16690732 TI - Results and feasibility of an external quality assessment scheme for self monitoring of blood glucose. AB - BACKGROUND: The analytical quality of self-monitoring of blood glucose (SMBG) is not satisfactory, and the need for standardized control routines for SMBG has been underscored. The objective of this study was to investigate whether an external quality assessment scheme (EQAS) designed for office laboratories could improve the quality of SMBG measurements. METHODS: From October 2001 through March 2004, we conducted 6 glucose surveys for diabetes patients and coordinated them with an EQAS for office laboratories. Patients received 2 control samples by post twice a year. They measured each control sample in duplicate in accordance with written instructions, returned the results, and received an assessment of their analytical performance. Participants who got a poor evaluation were followed up by phone and were offered guidance. RESULTS: Participating in an EQA program over a period of 3 years decreased the percentage of poor results among diabetes patients significantly, from 11.2% to 1.9% in the first and last surveys, respectively. Between-participant CVs improved from 5.5% to 3.7% and were comparable to results from office laboratories. It was difficult to sort out factors contributing to quality improvement. CONCLUSIONS: Implementing a traditional EQAS among diabetes patients may improve the analytical quality of SMBG and could be convenient for motivated patients. Further evaluation of the clinical usefulness of implementing such a program is needed, however, and costs as well as limitations of current EQAS for glucose in general should be taken into account. PMID- 16690733 TI - Oxidized forms of glutathione in peripheral blood as biomarkers of oxidative stress. AB - BACKGROUND: Reduced glutathione (GSH) and its redox forms, glutathione disulfide (GSSG) and glutathionylated proteins (PSSG), are biomarkers of oxidative stress, but methodologic artifacts can interfere with their measurement. We evaluated the importance of correct sample handling during the preanalytical phase for GSH, GSSG, and PSSG measurement. METHODS: We used human blood for in vitro experiments with oxidants [tert-butylhydroperoxide (t-BOOH), diamide, and menadione]. For in vivo experiments, we used rats in which we cannulated the jugular and femoral veins for both oxidant administration and blood collection. We measured GSH, GSSG, and PSSG with HPLC with or without sample pretreatment with N ethylmaleimide (NEM) to prevent artifacts. We also measured malondialdehyde (MDA) with HPLC, and protein carbonyls (PCO) with spectrophotometric procedures. RESULTS: When methodologic artifacts were prevented by pretreatment with NEM, GSSG results increased up to 3-fold over the basal concentrations, even in the presence of 5 micromol/L t-BOOH or diamide and 20 micromol/L menadione. PSSG increased by approximately 50% at 20 micromol/L t-BOOH or diamide and at 50 micromol/L menadione. PCO and MDA remained unchanged. In vivo oxidation treatments elicited immediate and significant increases in GSSG and PSSG over basal values (up to 200-fold), whereas PCO and MDA showed only slight variation 120 or 180 min after treatment. CONCLUSIONS: With the use of artifact-free measurement methods, GSH, GSSG, and PSSG are potentially powerful and reliable biomarkers of oxidative stress status and can be used to evaluate whether, and to what extent, oxidative stress may be involved in various diseases. PMID- 16690734 TI - Molecular diagnosis of Prader-Willi and Angelman syndromes by methylation specific melting analysis and methylation-specific multiplex ligation-dependent probe amplification. AB - BACKGROUND: Approximately 99% of Prader-Willi syndrome (PWS) and 80% of Angelman syndrome (AS) cases have deletions at a common region in chromosome 15q11.2-q13, uniparental disomy for chromosome 15 (UPD15), or imprinting center defects affecting gene expression in this region. The resulting clinical phenotype (PWS or AS) in each class of genomic abnormalities depends on the parent of origin. Both disorders are characterized at the molecular level by abnormal methylation of imprinted regions at 15q11.2-q13. Other rare chromosome 15 rearrangements and a few smaller atypical deletions associated with abnormal methylation patterns also have symptoms overlapping with either PWS or AS. METHODS: We designed a methylation-specific melting analysis (MS-MA) method for a rapid screening of PWS/AS and evaluated methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) for diagnosis of PWS/AS associated with deletions, UPD15, or rare duplications. Forty-nine previously genotyped samples were tested by MS MA. We also tested 26 MS-MA genotyped samples and 1 additional sample with rare duplication of chromosome region 15q11-q12. RESULTS: PWS/AS genotyping results obtained by MS-MA and by MS-MLPA were fully concordant. In addition, MS-MLPA was superior in detecting deletions/rare duplications, possible UPD15, or imprinting center defects, which were usually determined by a laborious fluorescence in situ hybridization method or by chromosomal segregation analysis for the parental origin using short-tandem repeat makers. CONCLUSIONS: MS-MA appears to be an efficient primary method to diagnose PWS/AS, and use of the quantitative MS-MLPA method provides detailed information about deletions, rare duplications, and possibly UPD. PMID- 16690735 TI - Development and validation of a multiplex add-on assay for sepsis biomarkers using xMAP technology. AB - BACKGROUND: Sepsis is a common and often fatal disease. Because sepsis can be caused by many different organisms, biomarkers that can aid in diagnosing sepsis and monitoring treatment efficacy are highly warranted. New sepsis markers may provide additional information to complement the currently used markers. METHODS: We used a combination of in-house and commercially available multiplex immunoassays based on Luminex xMAP technology to assay biomarkers of potential interest in EDTA-plasma samples. RESULTS: A 3-plex assay for soluble urokinase plasminogen activator receptor (suPAR), soluble triggering receptor expressed on myeloid cells-1 (sTREM-1), and macrophage migration inhibiting factor (MIF) was developed and validated in-house. This 3-plex assay was added to a commercially available interleukin-1beta (IL-1beta), IL-6, IL-8, granulocyte/macrophage colony stimulating factor, and tumor necrosis factor-alpha human cytokine panel. No cross-reactivity was observed when the assays were combined. Correlation between values obtained with the 8-plex, the 5-cytokine panel, the 3 in-house 1-plex assays, and a suPAR ELISA ranged from 0.86 to 0.99. Mean within- and between-run CVs were 8.0% and 11%, respectively. Recoveries of suPAR, sTREM-1, and MIF calibrators were 108%, 88%, and 51%, respectively. In plasma collected from 10 patients with bacterial sepsis confirmed by blood culture, the assay detected significantly increased concentrations of all 8 analytes compared with healthy controls. CONCLUSIONS: A commercially available xMAP panel can be expanded with markers of interest. The combined multiplex assay can measure the 8 analytes with high reproducibility. The xMAP technology is an appealing tool for assaying conventional cytokines in combination with new markers. PMID- 16690736 TI - Predictors of vitamin B6 and folate concentrations in older persons: the InCHIANTI study. AB - BACKGROUND: Low dietary intake and low serum concentrations of vitamin B6 and/or folate are associated with increased risk of vascular events, possibly because of their association with inflammation, which plays a crucial role in the pathogenesis of cardiovascular diseases. METHODS: Using data from 1320 participants in the population-based InCHIANTI study (586 men and 734 women; median age, 69 years; range, 21-102 years) for whom complete data on folate, vitamin B6, inflammatory markers, 5,10-methylenetetrahydrofolate reductase (MTHFR) C677T sequence variant, and important covariates were available, we evaluated the association of inflammatory markers with circulating concentrations of vitamin B6 and folate, independently of dietary vitamin intake, circulating vitamin concentrations, and MTHFR C677T sequence variant. RESULTS: According to multiple linear regression analysis, C-reactive protein and interleukin-6 receptor were strongly and negatively associated with circulating vitamin B6 but not with folate concentrations, independent of age, sex, serum creatinine, serum albumin, total energy intake, smoking history, dietary nutrient intake, and circulating homocysteine and vitamin concentrations. Serum folate concentrations were related to MTHFR 677 TT genotype in persons with folate intake in the lowest tertile (< 221.2 microg/day). Vitamin C and retinol intakes were strongly and positively associated with serum folate concentrations independent of age, sex, serum creatinine, serum albumin, total energy intake, smoking history, homocysteine plasma concentrations, dietary nutrient intakes, serum vitamin B6 and vitamin B12 concentrations, and MTHFR C677T sequence variant. CONCLUSIONS: Low serum vitamin B6, but not serum folate, concentrations are independent correlates of the proinflammatory state, and both are influenced by antioxidant reserves. PMID- 16690737 TI - Performance evaluation and cross-reactivity from insulin analogs with the ARCHITECT insulin assay. AB - BACKGROUND: Insulin measurement is used for the diagnosis of hypoglycemia and for insulin pharmacokinetic evaluations. We assessed the analytical and clinical performance of the ARCHITECT insulin assay, a chemiluminescent immunoassay recently introduced for the ARCHITECT i2000 fully automated immunoassay analyzer (Abbott Laboratories). We also tested whether major insulin analogs cross-reacted with the immunoassay reagents. METHODS: We used Clinical and Laboratory Standards Institute protocols to assess the analytical performance of the ARCHITECT insulin assay and compared its accuracy with that of the E-test TOSOH II (IRI) from TOSOH Corporation. We used 3 recombinant insulin analogs (lispro, aspart, and glargine) to evaluate the cross-reactivity of insulin analogs with the ARCHITECT immunoassay reagent. RESULTS: The total CV for the ARCHITECT assay was < 5%. Correlation between the ARCHITECT insulin assay and the E-test TOSOH II (IRI) was satisfactory in the measured range, but we detected a slope deviation between the assays. The ARCHITECT insulin assay showed low cross-reactivity to the insulin analog aspart, whereas it detected the other insulin analogs, lispro and glargine, in concentrations as high as the theoretical concentrations. CONCLUSIONS: The ARCHITECT insulin assay showed favorable basic performance, including reproducibility, dilution linearity, detection limit, and effects of interfering substances. When interpreting results, clinicians and laboratory pathologists should be aware of the cross-reactivity of the ARCHITECT and other immunoassays to specific insulin analogs prescribed to diabetes patients. PMID- 16690738 TI - Clinical-scale high-throughput analysis of urinary 8-oxo-7,8-dihydro-2' deoxyguanosine by isotope-dilution liquid chromatography-tandem mass spectrometry with on-line solid-phase extraction. AB - BACKGROUND: Quantification of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) in urine or blood is used to assess and monitor oxidative stress in patients. We describe the use of on-line solid-phase extraction (SPE) and isotope-dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS) for automated measurement of urinary 8-oxodGuo. METHODS: Automated purification of urine was accomplished with a switching valve and an Inertsil ODS-3 column. After the addition of 15N5-labeled 8-oxodGuo as an internal standard, urine samples were analyzed within 10 min without sample purification. This method was applied to measure urinary 8-oxodGuo in a group of healthy persons (32 regular smokers and 35 nonsmokers). Urinary cotinine was also assayed by an isotope-dilution LC-MS/MS method. RESULTS: The lower limit of detection was 5.7 ng/L on column (2.0 fmol). Inter- and intraday imprecision (CV) was < 5.0%. Mean recovery of 8-oxodGuo in urine was 99%-102%. Mean (SD) urinary concentrations of 8-oxodGuo in smokers [7.26 (3.14) microg/g creatinine] were significantly higher than those in nonsmokers [4.69 (1.70) microg/g creatinine; P < 0.005]. Urinary concentrations of 8-oxodGuo were significantly correlated with concentrations of cotinine in smokers (P < 0.05). CONCLUSIONS: This on-line SPE LC-MS/MS method is sufficiently sensitive, precise, and rapid to provide high-throughput direct analysis of urinary 8-oxodGuo without compromising quality and validation criteria. This method could be applicable for use in daily clinical practice for assessing oxidative stress in patients. PMID- 16690739 TI - Utility of N-terminal pro-B-type natriuretic peptide to differentiate cardiac diseases from noncardiac diseases in young pediatric patients. AB - BACKGROUND: Previous studies comparing children with cardiac disease with children with lung disease or healthy children indicated that natriuretic peptides are promising markers in pediatric patients. The aim of this study was to further clarify the diagnostic usefulness of N-terminal pro-B-type natriuretic peptide (NT-proBNP) measurements in a less preselected population of children younger than 3 years, a population in which clinical symptoms are frequently unspecific. METHODS: NT-proBNP concentrations (Roche Diagnostics) were measured in sera of 142 pediatric patients (age range, 33-1070 days) presenting at the Gynaecologic and Pediatric Hospital (Linz, Austria) between January 2003 and January 2004. ROC curve analysis for the diagnostic performance of NT-proBNP, the Mann-Whitney U-test for group comparison, and linear regression analysis for influencing factors were performed. RESULTS: NT-proBNP concentrations were significantly increased in infants with cardiac diseases [median (25th-75th percentile), 3681 (1045-13557) ng/L; n = 23] compared with infants with other diseases [241 (116-542) ng/L; n = 119], and ROC analysis revealed good performance for NT-proBNP in differentiating between infants with and without cardiac diseases [mean area under the curve (AUC) with 95% confidence interval (CI), 0.87 (0.76-0.94)]. A subgroup analysis of exactly age- and sex-matched infants was performed, which revealed results comparable to those for the whole study population [mean (95% CI) AUC, 0.84 (0.68-0.93)]. CONCLUSION: In a heterogeneous group of pediatric patients < 3 years of age, NT-proBNP showed good diagnostic performance to distinguish between cardiac diseases and various noncardiac diseases. PMID- 16690740 TI - The callipyge mutation enhances bidirectional long-range DLK1-GTL2 intergenic transcription in cis. AB - The callipyge mutation (CLPG) is an A to G transition that affects a muscle specific long-range control element located in the middle of the 90-kb DLK1-GTL2 intergenic (IG) region. It causes ectopic expression of a 327-kb cluster of imprinted genes in skeletal muscle, resulting in the callipyge muscular hypertrophy and its non-Mendelian inheritance pattern known as polar overdominance. We herein demonstrate that the CLPG mutation alters the muscular epigenotype of the DLK1-GTL2 IG region in cis, including hypomethylation, acquisition of novel DNase-I hypersentivite sites, and, most strikingly, strongly enhanced bidirectional, long-range IG transcription. The callipyge phenotype thus emerges as a unique model to study the functional significance of IG transcription, which recently has proven to be a widespread, yet elusive, feature of the mammalian genome. PMID- 16690741 TI - Toward the structural genomics of complexes: crystal structure of a PE/PPE protein complex from Mycobacterium tuberculosis. AB - The developing science called structural genomics has focused to date mainly on high-throughput expression of individual proteins, followed by their purification and structure determination. In contrast, the term structural biology is used to denote the determination of structures, often complexes of several macromolecules, that illuminate aspects of biological function. Here we bridge structural genomics to structural biology with a procedure for determining protein complexes of previously unknown function from any organism with a sequenced genome. From computational genomic analysis, we identify functionally linked proteins and verify their interaction in vitro by coexpression/copurification. We illustrate this procedure by the structural determination of a previously unknown complex between a PE and PPE protein from the Mycobacterium tuberculosis genome, members of protein families that constitute approximately 10% of the coding capacity of this genome. The predicted complex was readily expressed, purified, and crystallized, although we had previously failed in expressing individual PE and PPE proteins on their own. The reason for the failure is clear from the structure, which shows that the PE and PPE proteins mate along an extended apolar interface to form a four-alpha-helical bundle, where two of the alpha-helices are contributed by the PE protein and two by the PPE protein. Our entire procedure for the identification, characterization, and structural determination of protein complexes can be scaled to a genome-wide level. PMID- 16690742 TI - Counterions between charged polymers exhibit liquid-like organization and dynamics. AB - Current understanding of electrostatics in water is based on mean-field theories like the Poisson-Boltzmann formalism and its approximations, which are routinely used in colloid science and computational biology. This approach, however, breaks down for highly charged systems, which exhibit counterintuitive phenomena such as overcharging and like-charge attraction. Models of counterion correlations have been proposed as possible explanations, but no experimental comparisons are available. Here, collective dynamics of counterions that mediate like-charge attraction between F-actin filaments have been directly observed in aqueous solution using high-resolution inelastic x-ray scattering down to molecular length-scales. We find a previously undescribed acoustic-like phonon mode associated with correlated counterions. The excitation spectra at high wave vector Q reveal unexpected dynamics due to ions interacting with their "cages" of nearest neighbors. We examine this behavior in the context of intrinsic charge density variations on F-actin. The measured speed of sound and collective relaxation rates in this liquid agree surprisingly well with simple model calculations. PMID- 16690743 TI - Activation of FIP1L1-PDGFRalpha requires disruption of the juxtamembrane domain of PDGFRalpha and is FIP1L1-independent. AB - Genetic abnormalities that result in expression of chimeric tyrosine kinase proteins such as BCR-ABL1 and ETV6-PDGFRbeta are common causes of hematopoietic malignancies. The paradigm for constitutive activation of these fusion tyrosine kinases is enforced homodimerization by self-association domains present in the fusion partner proteins. The unique interstitial deletion on chromosome 4q12 that leads to expression of the FIP1L1-PDGFRalpha fusion tyrosine kinase was recently identified as a cause of chronic eosinophilic leukemia. In this report, we demonstrate that FIP1L1 is completely dispensable for PDGFRalpha activation in vitro and in vivo. Instead, truncation of PDGFRalpha between two conserved tryptophan residues in the juxtamembrane (JM) domain is required for kinase activation and transforming potential of FIP1L1-PDGFRalpha. The presence of a complete JM domain in FIP1L1-PDGFRalpha is inhibitory, but this autoinhibition can be overcome by enforced homodimerization. Similar effects of the JM domain in the context of PDGFRbeta were observed. These results suggest that disruption of the autoinhibitory JM domain is an alternative, dimerization-independent mechanism by which chimeric tyrosine kinases are constitutively activated and induce leukemogenesis. PMID- 16690744 TI - Spin glasses and fragile glasses: statics, dynamics, and complexity. AB - In this paper I will briefly review some theoretical results that have been obtained in recent years for spin glasses and fragile glasses. I will concentrate my attention on the predictions coming from the so called broken replica symmetry approach and on their experimental verifications. I will also mention the relevance or these results for other fields, and in general for complex systems. PMID- 16690745 TI - Oogenesis requires germ cell-specific transcriptional regulators Sohlh1 and Lhx8. AB - Mammalian oogenesis requires oocyte-specific transcriptional regulators. The full complement of oocyte-specific transcription factors is unknown. Here, we describe the finding that Sohlh1, a spermatogenesis and oogenesis basic helix-loop-helix transcription factor in females, is preferentially expressed in oocytes and required for oogenesis. Sohlh1 disruption perturbs follicular formation in part by causing down-regulation of two genes that are known to disrupt folliculogenesis: newborn ovary homeobox gene (Nobox) and factor in the germ-line alpha (Figla). In addition, we show that Lhx8 is downstream of Sohlh1 and critical in fertility. Thus, Sohlh1 and Lhx8 are two germ cell-specific, critical regulators of oogenesis. PMID- 16690746 TI - Fluorescence resonance energy transfer in living cells reveals dynamic membrane changes in the initiation of B cell signaling. AB - B cell responses are initiated by the clustering of the B cell receptor (BCR) by the binding of multivalent antigens. Clustering leads to phosphorylation of tyrosines in the cytoplasmic domains of the BCR by the inner plasma membrane leaflet-associated Src-family kinase Lyn. At present, little is known about the earliest events after BCR clustering that precede the BCR's phosphorylation by Lyn. Here we use fluorescence resonance energy transfer (FRET) in living cells to detect the interaction of the BCR with a Lyn-based membrane-targeted reporter in the first several seconds after BCR clustering. The results showed that, within seconds of antigen binding, the BCR selectively and transiently associated with the Lyn construct and that this association preceded by several seconds the triggering of Ca2+ fluxes and could be prolonged by the engagement of the B cell coreceptor complex, CD19/CD21. Thus, FRET measurements in living B cells revealed highly dynamic and regulated antigen-induced changes in the plasma membrane, allowing association of the BCR with the earliest components of its signaling cascade. PMID- 16690748 TI - Selenoprotein deficiency accelerates prostate carcinogenesis in a transgenic model. AB - Considerable animal and human data have indicated that selenium is effective in reducing the incidence of several different types of cancer, including that of the prostate. However, the mechanism by which selenium inhibits carcinogenesis remains unknown. One possibility is that dietary selenium influences the levels of selenium-containing proteins, or selenoproteins. Selenoproteins contain selenium in the form of selenocysteine and perform a variety of cellular functions, including antioxidant defense. To determine whether the levels of selenoproteins can influence carcinogenesis independent of selenium intake, a unique mouse model was developed by breeding two transgenic animals: mice with reduced selenoprotein levels because of the expression of an altered selenocysteine-tRNA (i6A-) and mice that develop prostate cancer because of the targeted expression of the SV40 large T and small t oncogenes to that organ [C3(1)/Tag]. The resulting bigenic animals (i6A-/Tag) and control WT/Tag mice were assessed for the presence, degree, and progression of prostatic epithelial hyperplasia and nuclear atypia. The selenoprotein-deficient mice exhibited accelerated development of lesions associated with prostate cancer progression, implicating selenoproteins in cancer risk and development and raising the possibility that selenium prevents cancer by modulating the levels of these selenoproteins. PMID- 16690749 TI - Glucocorticoid receptor activation signals through forkhead transcription factor 3a in breast cancer cells. AB - Activation of the glucocorticoid receptor (GR) plays a critical role in the stress response of virtually all cell types. Despite recent advances in large scale genomic and proteomic data acquisition, identification of physiologically relevant molecular events downstream of nuclear hormone receptor activation remains challenging. By analyzing gene expression changes 30 min after dexamethasone (Dex) treatment, we previously found that immediate induction of serum and glucocorticoid-regulated kinase-1 (SGK-1) expression is required for GR mediated mammary epithelial cell survival signaling. We now report that activation of the GR mediates Forkhead transcription factor 3a (FOXO3a) phosphorylation and inactivation in mammary epithelial cells. GR-mediated induction of SGK-1 expression is required for FOXO3a inactivation; additional growth factor stimulation is not required. To further explore the gene expression changes that occur downstream of GR-mediated FOXO3a inactivation, we analyzed temporal gene expression data and selected GR-down-regulated genes containing core FOXO3a binding motifs in their proximal promoters. This approach revealed several previously unrecognized transcriptional target genes of FOXO3a, including IGF binding protein-3 (IGFBP-3). Endogenous IGFBP-3 expression was confirmed to be dependent on the GR-SGK-1-FOXO3a signaling pathway. Moreover, GR activation decreased FOXO3a-induced apoptosis in SK-BR-3 breast cancer cells. Collectively, our data suggest that GR-mediated FOXO3a inactivation is an important mechanism contributing to glucocorticoid-mediated mammary epithelial cell survival. PMID- 16690747 TI - The origin of European cattle: evidence from modern and ancient DNA. AB - Cattle domestication from wild aurochsen was among the most important innovations during the Neolithic agricultural revolution. The available genetic and archaeological evidence points to at least two major sites of domestication in India and in the Near East, where zebu and the taurine breeds would have emerged independently. Under this hypothesis, all present-day European breeds would be descended from cattle domesticated in the Near East and subsequently spread during the diffusion of herding and farming lifestyles. We present here previously undescribed genetic evidence in contrast with this view, based on mtDNA sequences from five Italian aurochsen dated between 7,000 and 17,000 years B.P. and >1,000 modern cattle from 51 breeds. Our data are compatible with local domestication events in Europe and support at least some levels of introgression from the aurochs in Italy. The distribution of genetic variation in modern cattle suggest also that different south European breeds were affected by introductions from northern Africa. If so, the European cattle may represent a more variable and valuable genetic resource than previously realized, and previous simple hypotheses regarding the domestication process and the diffusion of selected breeds should be revised. PMID- 16690750 TI - Protein disulfide isomerase serves as a molecular chaperone to maintain estrogen receptor alpha structure and function. AB - The effects of the steroid hormone 17beta-estradiol are mediated through its interaction with the nuclear estrogen receptor (ER). Upon binding 17beta estradiol, the ER initiates changes in gene expression through its interaction with specific DNA sequences, estrogen response elements (EREs), and recruits coregulatory proteins that influence gene expression. To better understand how estrogen-responsive genes are regulated, we have isolated and identified proteins associated with ERalpha when it is bound to the consensus ERE. One of these proteins, protein disulfide isomerase (PDI), has two distinct functions: acting as a molecular chaperone to maintain properly folded proteins and regulating the redox state of proteins by catalyzing the thiol-disulfide exchange reaction through two thioredoxin-like domains. Using a battery of biochemical and molecular techniques, we have demonstrated that PDI colocalizes with ERalpha in MCF-7 nuclei, alters ERalpha conformation, enhances the ERalpha-ERE interaction in the absence and presence of an oxidizing agent, influences the ability of ERalpha to mediate changes in gene expression, and associates with promoter regions of two endogenous estrogen-responsive genes. Our studies suggest that PDI plays a critical role in estrogen responsiveness by functioning as a molecular chaperone and assisting the receptor in differentially regulating target gene expression. PMID- 16690751 TI - Psychological adjustment in children and families living with HIV. AB - OBJECTIVE: To assess psychological adjustment in children living with human immunodeficiency virus (HIV) and their primary caregivers. METHODS: The study protocol included use of standardized questionnaires to assess emotional and behavioral health of 57 children and 54 caregivers (Phase 1). Positive screening led to standardized interviews to assess current psychiatric diagnoses (Phase 2). RESULTS: Of the 16 children who entered Phase 2, 6 (38%) met the criteria for a psychiatric diagnosis. Of the 15 adults who met the screening criteria, 13 completed a computerized psychiatric interview and all 13 (100%) met the criteria for a psychiatric diagnosis. CONCLUSIONS: While important mental health needs were identified in families with HIV, the majority of families did not exhibit mental health disorders. These results might reflect the substantial psychosocial resilience of these families. Further study is needed to determine to what extent the mental health needs of children and their caregivers are being met. In addition, identification of protective factors in resilience and coping in families living with a chronic illness is warranted. PMID- 16690752 TI - Prevalence and correlates of early onset asthma and wheezing in a healthy birth cohort of 2- to 3-year olds. AB - OBJECTIVE: The combined contribution of neonatal, perinatal, and maternal health, demographic, environmental, and family psychosocial factors to early onset asthma and wheezing in a healthy birth cohort was examined. METHODS: Participants included 1,158 ethnically and socioeconomically diverse parents of 2- and 3-year olds who completed mailed questionnaires. RESULTS: Asthma and wheezing prevalence was 8.4 and 8.1%, respectively. Asthma during pregnancy, smoking in the home, and being male increased risk for asthma diagnosis and wheezing whereas social support minimized risk for both. Shorter gestational age, exposure to violence, and maternal anxiety increased risk for wheezing. The negative impact of smoking in the home was greatest for children with shorter gestational ages and mothers with asthma during pregnancy. CONCLUSION: Findings confirm and extend previous work documenting demographic risks and highlight smoking, violent events, and social support in early onset asthma and wheezing. Findings illustrate the need for ecologically based interventions to treat asthma and wheezing in young children. PMID- 16690753 TI - Identification and functional validation of novel autoantigens in equine uveitis. AB - The development, progression, and recurrence of autoimmune diseases are frequently driven by a group of participatory autoantigens. We identified and characterized novel autoantigens by analyzing the autoantibody binding pattern from horses affected by spontaneous equine recurrent uveitis to the retinal proteome. Cellular retinaldehyde-binding protein (cRALBP) had not been described previously as autoantigen, but subsequent characterization in equine recurrent uveitis horses revealed B and T cell autoreactivity to this protein and established a link to epitope spreading. We further immunized healthy rats and horses with cRALBP and observed uveitis in both species with typical tissue lesions at cRALBP expression sites. The autoantibody profiling outlined here could be used in various autoimmune diseases to detect autoantigens involved in the dynamic spreading cascade or serve as predictive markers. PMID- 16690754 TI - dILA neurons in the dorsal spinal cord are the product of terminal and non terminal asymmetric progenitor cell divisions, and require Mash1 for their development. AB - dILA and dILB neurons comprise the major neuronal subtypes generated in the dorsal spinal cord, and arise in a salt-and-pepper pattern from a broad progenitor domain that expresses the bHLH factor Mash1. In this domain, Mash1 positive and Mash1-negative cells intermingle. Using a Mash1(GFP) allele in mice, we show here that Mash1+ progenitors give rise to dILA and dILB neurons. Using retroviral tracing in the chick, we demonstrate that a single progenitor can give rise to a dILA and a dILB neuron, and that dILA neurons are the product of asymmetric progenitor cell divisions. In Mash1-null mutant mice, the development of dILA, but not of dILB neurons is impaired. We provide evidence that a dual function of Mash1 in neuronal differentiation and specification accounts for the observed changes in the mutant mice. Our data allow us to assign to Mash1 a function in asymmetric cell divisions, and indicate that the factor coordinates cell cycle exit and specification in the one daughter that gives rise to a dILA neuron. PMID- 16690755 TI - Robo1 regulates the development of major axon tracts and interneuron migration in the forebrain. AB - The Slit genes encode secreted ligands that regulate axon branching, commissural axon pathfinding and neuronal migration. The principal identified receptor for Slit is Robo (Roundabout in Drosophila). To investigate Slit signalling in forebrain development, we generated Robo1 knockout mice by targeted deletion of exon 5 of the Robo1 gene. Homozygote knockout mice died at birth, but prenatally displayed major defects in axon pathfinding and cortical interneuron migration. Axon pathfinding defects included dysgenesis of the corpus callosum and hippocampal commissure, and abnormalities in corticothalamic and thalamocortical targeting. Slit2 and Slit1/2 double mutants display malformations in callosal development, and in corticothalamic and thalamocortical targeting, as well as optic tract defects. In these animals, corticothalamic axons form large fasciculated bundles that aberrantly cross the midline at the level of the hippocampal and anterior commissures, and more caudally at the medial preoptic area. Such phenotypes of corticothalamic targeting were not observed in Robo1 knockout mice but, instead, both corticothalamic and thalamocortical axons aberrantly arrived at their respective targets at least 1 day earlier than controls. By contrast, in Slit mutants, fewer thalamic axons actually arrive in the cortex during development. Finally, significantly more interneurons (up to twice as many at E12.5 and E15.5) migrated into the cortex of Robo1 knockout mice, particularly in both rostral and parietal regions, but not caudal cortex. These results indicate that Robo1 mutants have distinct phenotypes, some of which are different from those described in Slit mutants, suggesting that additional ligands, receptors or receptor partners are likely to be involved in Slit/Robo signalling. PMID- 16690756 TI - Commentary: economic crisis or structural adjustment--which is worse for child health in African countries? PMID- 16690757 TI - Commentary: the associations between height, cognition, and education and their relevance for health studies. PMID- 16690758 TI - Haplotype analysis revealed no association between the PTPN22 gene and RA in a Japanese population. AB - OBJECTIVE: The protein tyrosine phosphatase non-receptor type 22 (PTPN22) gene is a member of the PTPs that negatively regulate T-cell activation. A missense single nucleotide polymorphism (SNP) in the PTPN22 gene known as R620W was recently reported to be associated with several autoimmune diseases including rheumatoid arthritis (RA). The association was confirmed repeatedly in the populations of North European ancestry. However, the SNP was reported to be non polymorphic in the Asian populations. Because the gene confers an impact on autoimmune diseases, we attempt to explore an association between PTPN22 gene and RA in a Japanese population without restricting to the SNP, R620W. METHODS: We studied 1128 RA patients and 455 controls. In addition to the SNP, R620W, we selected eight testing SNPs spanning 45 kb over the PTPN22 gene using the International HapMap Project. Genotyping was performed using the TaqMan fluorogenic 5' nuclease assay. Associations between RA and each of the SNPs were estimated by the Fisher's exact test. Haplotype was constructed using the expectation-maximization algorithm. RESULTS: R620W was not polymorphic enough in both the patients and the controls, and was therefore excluded from further analysis. Each allele frequency for the eight other SNPs in both groups was compared and no association was detected. Haplotype analysis also revealed that PTPN22 gene was not associated with RA in a Japanese population. CONCLUSION: We found no association between PTPN22 and RA in a Japanese population. The result suggests that the PTPN22 gene is associated with RA only in a specific ethnic group. PMID- 16690759 TI - Therapy-resistent lupus skin disease successfully treated with rituximab. PMID- 16690760 TI - Interleukin-6 signalling in juvenile idiopathic arthritis is limited by proteolytically cleaved soluble interleukin-6 receptor. AB - OBJECTIVES: Interleukin-6 (IL-6) exerts multiple effects on chondrocytes and fibroblasts within the joint and is associated with disease activity in juvenile idiopathic arthritis (JIA). Although these cells express the ubiquitous signalling receptor for all IL-6-related cytokines, gp130, they do not express a cognate IL-6 receptor. Consequently, IL-6 responses within these cells occur via IL-6 trans-signalling relying on the presence of a soluble receptor (sIL-6R). Levels of sIL-6R in vivo are governed by either proteolytic cleavage (PC) of cognate receptor or by differential sIL-6R mRNA splicing (DS). The aim of this study was to evaluate the contribution of both isoforms to clinical parameters associated with IL-6 signalling in JIA. METHODS: IL-6, sIL-6R and DS-sIL-6R were measured by ELISA in serum and synovial fluid (SF) samples from 86 JIA patients. These data were related to indicators of inflammation-erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) and compared between patients stratified by subtype, age and disease duration. RESULTS: SF IL-6 significantly correlated with general indicators of activity (ESR and CRP) and SF PC-sIL-6R to a lesser degree with CRP. When the IL-6:sIL-6R ratio was calculated as an indicator of the potential for IL-6 signalling within the joint, 33% of SF samples showed a ratio >1 indicating saturation of sIL-6R by IL-6. Mean DS-sIL-6R levels were 0.71 ng/ml, whereas PC-sIL-6R levels constituted the majority of sIL-6R at 20.89 ng/ml. CONCLUSIONS: IL-6 trans-signalling within the joints of JIA patients is predominantly governed by the presence of PC-sIL-6R, and the data provided suggest that synovial levels of IL-6 and sIL-6R would be sufficient to drive IL-6 responses in chondrocytes and synovial fibroblasts. PMID- 16690761 TI - Against the current--when primary pericardial disease causes rheumatic disease. PMID- 16690762 TI - Are rheumatologists' treatment decisions influenced by patients' age? AB - OBJECTIVES: The objective of this study was to determine whether physicians' treatment preferences are influenced by patients' age. METHODS: We mailed a survey to a random sample of rheumatologists practicing in the US. The survey included a scenario describing a hypothetical patient with rheumatoid arthritis (RA) on hydroxychloroquine, sulfasalazine and low-dose prednisolone, who presents with active disease during a follow-up appointment. The scenario was formulated in two versions that were identical except for the age of the patient. After reading the scenario, respondents were asked to rate (on a 10 cm numerical rating scale) their recommendations for each of the three options: (i) increasing the dose of prednisolone, (ii) adding a new disease-modifying anti-rheumatic drug (DMARD) and (iii) switching DMARDs. Rheumatologists who rated either adding a new DMARD or switching DMARDs higher than increasing the dose of prednisolone were classified as 'preferring aggressive treatment with DMARDs', while the others were classified as 'NOT preferring aggressive treatment with DMARDs'. RESULTS: A total of 480 rheumatologists were mailed a questionnaire; 204 responded, giving a response rate of 42.5%. Overall 163 (80%) respondents were classified as preferring aggressive treatment with DMARDs. Rheumatologists responding to this survey were more likely to prefer aggressive DMARD treatment for the young RA patient vs the older RA patient (87 vs 71%, P= 0.007). CONCLUSIONS: Our findings suggest that rheumatologists' treatment recommendations may be influenced by age. Future educational efforts should increase physician awareness of this possible bias in order to ensure equal service delivery across ages. PMID- 16690763 TI - Prevalence and management of rheumatoid arthritis in the general population of Greece--the ESORDIG study. AB - OBJECTIVE: To assess the prevalence and management of rheumatoid arthritis (RA) in the general adult population of Greece. METHODS: This cross-sectional study was conducted on the total adult population (> or =19 yrs old) of seven communities (8,547 subjects), and on 2,100 out of 5,686 randomly selected subjects in two additional communities. The study, based on a standardized questionnaire and clinical evaluation and laboratory investigation when necessary, was carried out by rheumatologists who visited the target population at their homes. Diagnosis of RA was based on the American College of Rheumatology (ACR) 1987 criteria. RESULTS: A total of 8,740 subjects participated (response rate 82.1%). RA was diagnosed in 59 individuals. The prevalence of RA was 0.68% (95% CI 0.51-0.85); it was significantly higher in females than males (P< 0.0005), and increased significantly with age up to and including the 50-59-yr old group (P< 0.002), and then decreased slightly. On their first medical visit, 19% (95% CI 9.7-30.9) of the RA patients had consulted a rheumatologist, while during the first year after disease onset, 61% (95% CI 48.6-73.4) had done so. Early consultation with a rheumatologist and disease-modifying anti-rheumatic drug (DMARD) combination therapy were negatively associated with ACR functional classes II-IV [adjusted odds ratios 0.18 (95% CI 0.04-0.85) and 0.17 (95% CI 0.04 0.72), respectively]. CONCLUSIONS: The prevalence of RA in the general adult population of Greece is similar to that in many other European countries; early consultation with a rheumatologist and DMARD combination therapy are associated with a better RA outcome. PMID- 16690764 TI - Effects of acute hyperosmolality on blood-brain barrier function in ovine fetuses and lambs. AB - We examined the effects of hyperosmolality on blood-brain barrier (BBB) permeability during development to test the vulnerability of the immature barrier to stress. The BBB response to hyperosmolality was quantified using the blood-to brain transfer constant (Ki) with alpha-aminoisobutyric acid in fetuses at 60% and 90% gestation, premature, newborn, and older lambs. Ki plotted against osmolality increased as a function of increases in osmolality in all groups and brain regions. The relationship was described (P < 0.05) by a segmented regression model. At lower osmolalities, changes in Ki were minimal, but after a break point (threshold) was reached, the increase (P < 0.05) was linear. We examined the responses of Ki to hyperosmolality within each brain region by comparing the thresholds and slopes of the second regression segment. Lower thresholds and higher slopes imply greater vulnerability to hyperosmolality in the younger groups. Thresholds increased (P < 0.05) with development in the thalamus, superior colliculus, pons, and spinal cord, and slopes of the second regression segment decreased (P < 0.05) in the cerebellum, hippocampus, inferior colliculus, medulla, and spinal cord. BBB resistance to hyperosmolality increased (P < 0.05) with development in most brain regions. The pattern of the Ki plotted against osmolality was (P < 0.05) heterogenous among brain regions in fetuses and premature and newborn lambs, but not in older lambs. We conclude that 1) BBB permeability increased as a function of changes in osmolality, 2) the barrier becomes more resistant to hyperosmolality during development, and 3) the permeability response to hyperosmolality is heterogenous among brain regions in fetuses and premature and newborn lambs. PMID- 16690765 TI - Manipulation of dietary carbohydrates after prolonged effort modifies muscle sarcoplasmic reticulum responses in exercising males. AB - The hypothesis tested was that disturbances in the sarcoplasmic reticulum (SR) Ca2+-cycling responses to exercise would associate with muscle glycogen reserves. Ten untrained males [peak O2 consumption (VO2 peak) = 3.41 +/- 0.20 (SE) l/min] performed a standardized cycle test (approximately 70% VO2 peak) on two occasions, namely, following 4 days of a high (Hi CHO)- and 4 days of a low (Lo CHO)-carbohydrate diet. Both Hi CHO and Lo CHO were preceded by a session of prolonged exercise designed to deplete muscle glycogen. SR Ca2+ cycling in crude homogenates prepared from vastus lateralis samples indicated higher (P < 0.05) Ca2+ uptake (microM x g protein(-1) x min(-1)) in Hi CHO compared with Lo CHO at 30 min (2.93 +/- 0.10 vs. 2.23 +/- 0.12) and at 67 min (2.77 +/- 0.16 vs. 2.10 +/ 0.12) of exercise, the point of fatigue in Lo CHO. Similar effects (P < 0.05) were noted between conditions for maximal Ca2+-ATPase (microM x g protein(-1) x min(-1)) at 30 min (142 +/- 8.5 vs. 107 +/- 5.0) and at 67 min (130 +/- 4.5 vs. 101 +/- 4.7). Both phase 1 and phase 2 Ca2+ release were 23 and 37% higher (P < 0.05) at 30 min of exercise and 15 and 34% higher (P < 0.05), at 67 min during Hi CHO compared with Lo CHO, respectively. No differences between conditions were observed at rest for any of these SR properties. Total muscle glycogen (mmol glucosyl units/kg dry wt) was higher (P < 0.05) in Hi CHO compared with Lo CHO at rest (+36%), 30 min (+53%), and at 67 min (+44%) of cycling. These results indicate that exercise-induced reductions in SR Ca2+-cycling properties occur earlier in exercise during low glycogen states compared with high glycogen states. PMID- 16690766 TI - Menstrual cycle phase and sex influence muscle glycogen utilization and glucose turnover during moderate-intensity endurance exercise. AB - Numerous studies from our and other laboratories have shown that women have a lower respiratory exchange ratio (RER) during exercise than equally trained men, indicating a greater reliance on fat oxidation. Differences in estrogen concentration between men and women likely play a role in this sex difference. Differing estrogen and progesterone concentrations during the follicular (FP) and luteal (LP) phases of the female menstrual cycle suggest that fuel use may also vary between phases. The purpose of the current study was to determine the effect of menstrual cycle phase and sex upon glucose turnover and muscle glycogen utilization during endurance exercise. Healthy, recreationally active young women (n = 13) and men (n = 11) underwent a primed constant infusion of [6,6-2H]glucose with muscle biopsies taken before and after a 90-min cycling bout at 65% peak O2 consumption. LP women had lower glucose rate of appearance (Ra, P = 0.03), rate of disappearance (Rd, P = 0.03), and metabolic clearance rate (MCR, P = 0.04) at 90 min of exercise and lower proglycogen (P = 0.04), macroglycogen (P = 0.04), and total glycogen (P = 0.02) utilization during exercise compared with FP women. Men had a higher RER (P = 0.02), glucose Ra (P = 0.03), Rd (P = 0.03), and MCR (P = 0.01) during exercise compared with FP women, and men had a higher RER at 75 and 90 min of exercise (P = 0.04), glucose Ra (P = 0.01), Rd (P = 0.01), and MCR (P = 0.001) and a greater PG utilization (P = 0.05) compared with LP women. We conclude that sex, and to a lesser extent menstrual cycle, influence glucose turnover and glycogen utilization during moderate-intensity endurance exercise. PMID- 16690767 TI - Regulation of sodium-glucose cotransporter SGLT1 in the intestine of hypertensive rats. AB - Experimental models of hypertension, such as spontaneously hypertensive rats (SHR), show alterations in cellular sodium transport that affects Na(+)-coupled cotransport processes and has been involved in the pathogenesis of this disease. The objective of the present study was to analyze the kinetic properties of the sodium-dependent glucose transport in the jejunum and ileum of SHR and its genetic control, Wistar-Kyoto (WKY) rats, as well as the regulation of the transporter, SGLT1. In hypertensive rats, the increased systolic blood pressure was accompanied by an enhancement of serum aldosterone levels compared with WKY rats, but no alterations were found in their body weight or serum glucose/insulin levels. The values for d-glucose maximal rate of transport (V(max)) were 42 and 60% lower, respectively, in the jejunum and ileum of SHR than those from WKY rats. On the other hand, the values for the Michaelis constant (K(m)) were similar in both animal groups, as was the diffusive component of transport (K(d)). Immunoblotting and Northern blot analysis revealed the existence of a lower abundance of SGLT1 protein and mRNA in SHR. Moreover, hypertensive rats showed a decrease in the molecular mass of SGLT1 that could not be explained in terms of different glycosylation and/or phosphorylation levels or an alternative splicing in the expression of the protein. These findings demonstrate that SGLT1 is regulated at a transcriptional level in the intestine of hypertensive rats, and suggest that this transporter might participate in the dysregulation of sodium transport observed in hypertension. PMID- 16690769 TI - Comments on: "Esophageal-gastric relaxation reflex in rat: dual control of peripheral nitrergic and cholinergic transmission". PMID- 16690770 TI - Serotonin-3 receptors in gastric mechanisms of cholecystokinin-induced satiety. PMID- 16690771 TI - Illuminating serotonergic gateways for strong resetting of the mammalian circadian clock. PMID- 16690772 TI - Excessive erythrocytosis in adult mice overexpressing erythropoietin leads to hepatic, renal, neuronal, and muscular degeneration. AB - To investigate the consequences of inborn excessive erythrocytosis, we made use of our transgenic mouse line (tg6) that constitutively overexpresses erythropoietin (Epo) in a hypoxia-independent manner, thereby reaching hematocrit levels of up to 0.89. We detected expression of human Epo in the brain and, to a lesser extent, in the lung but not in the heart, kidney, or liver of tg6 mice. Although no acute cardiovascular complications are observed, tg6 animals have a reduced lifespan. Decreased swim performance was observed in 5-mo-old tg6 mice. At about 7 mo, several tg6 animals developed spastic contractions of the hindlimbs followed by paralysis. Morphological analysis by light and electron microscopy showed degenerative processes in liver and kidney characterized by increased vascular permeability, chronic progressive inflammation, hemosiderin deposition, and general vasodilatation. Moreover, most of the animals showed severe nerve fiber degeneration of the sciatic nerve, decreased number of neuromuscular junctions, and degeneration of skeletal muscle fibers. Most probably, the developing demyelinating neuropathy resulted in muscular degeneration demonstrated in the extensor digitorum longus muscle. Taken together, chronically increased Epo levels inducing excessive erythrocytosis leads to multiple organ degeneration and reduced life expectancy. This model allows investigation of the impact of excessive erythrocytosis in individuals suffering from polycythemia vera, chronic mountain sickness, or in subjects tempted to abuse Epo by means of gene doping. PMID- 16690773 TI - Adrenergic regulation of HSL serine phosphorylation and activity in human skeletal muscle during the onset of exercise. AB - Skeletal muscle hormone-sensitive lipase (HSL) activity is increased by contractions and increases in blood epinephrine (EPI) concentrations and cyclic AMP activation of the adrenergic pathway during prolonged exercise. To determine the importance of hormonal stimulation of HSL activity during the onset of moderate- and high-intensity exercise, nine men [age 24.3 +/- 1.2 yr, 80.8 +/- 5.0 kg, peak oxygen consumption (VO2 peak) 43.9 +/- 3.6 ml x kg(-1) x min(-1)] cycled for 1 min at approximately 65% VO2 peak, rested for 60 min, and cycled at approximately 90% VO2 peak for 1 min. Skeletal muscle biopsies were taken pre- and postexercise, and arterial blood was sampled throughout exercise. Arterial EPI increased (P < 0.05) postexercise at 65% (0.45 +/- 0.10 to 0.78 +/- 0.27 nM) and 90% VO2 peak (0.57 +/- 0.34 to 1.09 +/- 0.50 nM). HSL activity increased (P < 0.05) following 1 min of exercise at 65% VO2 peak [1.05 +/- 0.39 to 1.78 +/- 0.54 mmol x min(-1) x kg dry muscle (dm)(-1)] and 90% VO2 peak (1.07 +/- 0.24 to 1.91 +/- 0.62 mmol x min(-1) x kg dm(-1)). Cyclic AMP content also increased (P < 0.05) at both exercise intensities (65%: 1.52 +/- 0.67 to 2.75 +/- 1.12, 90%: 1.85 +/- 0.65 to 2.64 +/- 0.93 micromol/kg dm). HSL Ser660 phosphorylation (approximately 55% increase) and ERK1/2 phosphorylation ( approximately 33% increase) were augmented following exercise at both intensities, whereas HSL Ser563 and Ser565 phosphorylation were not different from rest. The results indicate that increases in arterial EPI concentration during the onset of moderate- and high-intensity exercise increase cyclic AMP content, which results in the phosphorylation of HSL Ser660. This adrenergic stimulation contributes to the increase in HSL activity that occurs in human skeletal muscle in the first minute of exercise at 65% and 90% VO2 peak. PMID- 16690774 TI - Regional differences in myosin heavy chain isoform expression and maximal shortening velocity of the rat vaginal wall smooth muscle. AB - Contractility of the proximal and distal vaginal wall smooth muscle may play distinct roles in the female sexual response and pelvic support. The goal of this study was to determine whether differences in contractile characteristics of smooth muscle from these regions reside in differences in the expression of isoforms of myosin, the molecular motor for muscle contraction. Adult female Sprague-Dawley rats were killed on the day of estrus, and the vagina was dissected into proximal and distal segments. The Vmax at peak force was greater for tissue strips of the proximal vagina compared with that of distal (P < 0.01), although, at steady state, the Vmax for the muscle strips from the two regions was not different. Furthermore, at steady state, muscle stress was higher (P < 0.001) for distal vaginal strips (n = 5). Consistent with the high Vmax for the proximal vaginal strips, RT-PCR results revealed a higher %SM-B (P < 0.001) in the proximal vagina. A greater expression of SM-B protein (P < 0.001) was also detected by Western blotting (n = 4). Interestingly, there was no regional difference noted in SM-1/SM-2 isoforms (n = 6). The proximal vagina had a higher expression of myosin heavy chain protein (P < 0.01) and a greater percentage of smooth muscle bundles (P < 0.001). The results of this study are the first demonstration of a regional heterogeneity in Vmax and myosin isoform distribution in the vagina wall smooth muscle and confirm that the proximal vaginal smooth muscle exhibits phasic contractile characteristics compared with the distal vaginal smooth muscle, which is tonic. PMID- 16690775 TI - Modification of cutaneous vasodilator response to heat stress by daytime exogenous melatonin administration. AB - In humans, the nocturnal fall in internal temperature is associated with increased endogenous melatonin and with a shift in the thermoregulatory control of skin blood flow (SkBF), suggesting a role for melatonin in the control of SkBF. The purpose of this study was to test whether daytime exogenous melatonin would shift control of SkBF to lower internal temperatures during heat stress, as is seen at night. Healthy male subjects (n = 8) underwent body heating with melatonin administration (Mel) or without (control), in random order at least 1 wk apart. SkBF was monitored at sites pretreated with bretylium to block vasoconstrictor nerve function and at untreated sites. Cutaneous vascular conductance, calculated from SkBF and arterial pressure, sweating rate (SR), and heart rate (HR) were monitored. Skin temperature was elevated to 38 degrees C for 35-50 min. Baseline esophageal temperature (Tes) was lower in Mel than in control (P < 0.01). The Tes threshold for cutaneous vasodilation and the slope of cutaneous vascular conductance with respect to Tes were also lower in Mel at both untreated and bretylium-treated sites (P < 0.05). The Tes threshold for the onset of sweating and the Tes for a standard HR were reduced in Mel. The slope of the relationship of HR, but not SR, to Tes was lower in Mel (P < 0.05). These findings suggest that melatonin affects the thermoregulatory control of SkBF during hyperthermia via the cutaneous active vasodilator system. Because control of SR and HR are also modified, a central action of melatonin is suggested. PMID- 16690776 TI - Ionic channels and conductance-based models for hypothalamic neuronal thermosensitivity. AB - Thermoregulatory responses are partially controlled by the preoptic area and anterior hypothalamus (PO/AH), which contains a mixed population of temperature sensitive and insensitive neurons. Immunohistochemical procedures identified the extent of various ionic channels in rat PO/AH neurons. These included pacemaker current channels [i.e., hyperpolarization-activated cyclic nucleotide-gated channels (HCN)], background potassium leak channels (TASK-1 and TRAAK), and transient receptor potential channel (TRP) TRPV4. PO/AH neurons showed dense TASK 1 and HCN-2 immunoreactivity and moderate TRAAK and HCN-4 immunoreactivity. In contrast, the neuronal cell bodies did not label for TRPV4, but instead, punctate labeling was observed in traversing axons or their terminal endings. On the basis of these results and previous electrophysiological studies, Hodgkin-Huxley-like models were constructed. These models suggest that most PO/AH neurons have the same types of ionic channels, but different levels of channel expression can explain the inherent properties of the various types of temperature-sensitive and insensitive neurons. PMID- 16690777 TI - Mouse models of hematopoietic engraftment: limitations of transgenic green fluorescent protein strains and a high-performance liquid chromatography approach to analysis of erythroid chimerism. AB - Transgenic mouse strains ubiquitously expressing green fluorescent protein (GFP) have enabled investigators to develop in vivo transplant models that can detect donor contributions to many different tissues. However, most GFP transgenics lack expression of the reporter in the erythroid lineage. We evaluated expression of GFP in the bone marrow of the OsbY01 transgenic mouse (B6-GFP) in the context of CD71 and TER-119 expression and found that GFP fluorescence is lost prior to the basophilic erythroblast stage of development. However, platelets in B6-GFP mice were found to be uniformly positive for GFP. We therefore used the GFP transgenic model in combination with allelic variants of CD45 and the hemoglobin beta (Hbb) chain to develop a model system that allows all blood lineages to be followed in a mouse model of bone marrow transplantation (BMT). To detect Hbb variant molecules, we developed a new protocol based on high-performance liquid chromatography that is sensitive and precise, allowing rapid and quantitative analysis of erythroid chimerism. Platelet and leukocyte engraftment were detected by flow cytometry. BMT into sublethally irradiated (4 Gy) recipients demonstrated the failure of B6-GFP-derived cells to engraft relative to B6-CD45(a)-derived cells, suggesting that an immune barrier may prevent efficient engraftment of the transgenic cells in a setting of minimal ablation. These results establish limitations in the use of transgenic GFP expression as a donor marker in transplantation models. PMID- 16690778 TI - Integrins are markers of human neural stem cells. AB - The identification of markers for the isolation of human neural stem cells (hNSCs) is essential for studies of their biology and therapeutic applications. This study investigated expression of the integrin receptor family by hNSCs as potential markers. Selection of alpha6(hi) or beta1(hi) cells by fluorescence activated cell sorting led to an enrichment of human neural precursors, as shown by both neurosphere forming assays and increased expression of prominin-1, sox2, sox3, nestin, bmi1, and musashi1 in the beta1(hi) population. Cells expressing high levels of beta1 integrin also expressed prominin-1 (CD133), a marker previously used to isolate hNSCs, and selection using integrin beta1(hi) cells or prominin-1(hi) cells was found to be equally effective at enriching for hNSCs from neurospheres. Therefore, integrin subunits alpha6 and beta1 are highly expressed by human neural precursors and represent convenient markers for their prospective isolation. PMID- 16690779 TI - Equivalency of nuclear transfer-derived embryonic stem cells to those derived from fertilized mouse blastocysts. AB - Therapeutic cloning, whereby nuclear transfer (NT) is used to generate embryonic stem cells (ESCs) from blastocysts, has been demonstrated successfully in mice and cattle. However, if NT-ESCs have abnormalities, such as those associated with the offspring produced by reproductive cloning, their scientific and medical utilities might prove limited. To evaluate the characteristics of NT-ESCs, we established more than 150 NT-ESC lines from adult somatic cells of several mouse strains. Here, we show that these NT-ESCs were able to differentiate into all functional embryonic tissues in vivo. Moreover, they were identical to blastocyst derived ESCs in terms of their expression of pluripotency markers in the presence of tissue-dependent differentially DNA methylated regions, in DNA microarray profiles, and in high-coverage gene expression profiling. Importantly, the NT procedure did not cause irreversible damage to the nuclei. These similarities of NT-ESCs and ESCs indicate that murine therapeutic cloning by somatic cell NT can provide a reliable model for preclinical stem cell research. PMID- 16690780 TI - Wnt signaling regulates the invasion capacity of human mesenchymal stem cells. AB - Human mesenchymal stem cells (hMSCs) exhibit the potential to contribute to a wide variety of endogenous organ tissue repair. However, the signals governing hMSC mobilization out of the bone marrow, release into the bloodstream, and migration/invasion into the target tissue are largely unknown. Since canonical Wnt signaling regulates not only tumor but also various stem cell attributes, we hypothesized that this signal transduction pathway might also be involved in governing the transmigration of hMSCs through human extracellular matrix (ECM). Stimulation of hMSCs with recombinant Wnt3a or LiCl resulted in the accumulation of the transcriptional activator beta-catenin, its translocation into the nucleus, and the upregulation of typical Wnt target genes such as cyclin D1 and membrane-type matrix metalloproteinase-1 (MT1-MMP). Moreover, both stimuli significantly enhanced hMSC proliferation up to 40%. In addition, an increase of more than twofold in the ability of hMSCs to transmigrate through Transwell filters coated with human ECM was observed. In a reverse approach, Wnt signaling in hMSCs was inhibited by knocking down the expression of either beta-catenin or low-density lipoprotein receptor-related protein 5 using RNA interference technology. These inhibition strategies resulted in downregulation of the Wnt target genes cyclin D1 and MT1-MMP, in a reduced proliferation rate, and in a strikingly diminished invasion capacity (64% and 52%). Taken together, this study provides for the first time decisive evidence that canonical Wnt signaling is critically involved in the regulation of the proliferation, as well as of the migration/invasion capacity of hMSCs, representing essential stem cell features indispensable during tissue regeneration processes. PMID- 16690781 TI - Quantitative Oct4 overproduction in mouse embryonic stem cells results in prolonged mesoderm commitment during hematopoietic differentiation in vitro. AB - The Oct4 transcription factor is essential for the self-renewal and pluripotency of embryonic stem cells (ESCs). Oct4 level also controls the fate of ESCs. We analyzed the effects of Oct4 overproduction on the hematopoietic differentiation of ESCs. Oct4 was introduced into ESCs via a bicistronic retroviral vector, and cells were selected on the basis of Oct4 production, with Oct4(+) and Oct4(2+) displaying twofold and three- to fourfold overproduction, respectively. Oct4 overproduction inhibited hematopoietic differentiation in a dose-dependent manner, after the induction of such differentiation by the formation of day 6 embryoid bodies (EB6). This effect resulted from defective EB6 formation rather than from defective hematopoietic differentiation. In contrast, when hematopoiesis was induced by the formation of blast colonies, the effects of Oct4 depended on the level of overproduction: twofold overproduction increased hematopoietic differentiation, whereas higher levels of overproduction markedly inhibited hematopoietic development. This increase or maintenance of Oct4 levels appears to alter the kinetics and pattern of mesoderm commitment, thereby modifying hemangioblast generation. These results demonstrate that Oct4 acts as a master regulator of ESC differentiation. PMID- 16690782 TI - Mechanisms of cardiolipin oxidation by cytochrome c: relevance to pro- and antiapoptotic functions of etoposide. AB - Execution of apoptotic program in mitochondria is associated with accumulation of cardiolipin peroxidation products required for the release of proapoptotic factors into the cytosol. This suggests that lipid antioxidants capable of inhibiting cardiolipin peroxidation may act as antiapoptotic agents. Etoposide, a widely used antitumor drug and a topoisomerase II inhibitor, is a prototypical inducer of apoptosis and, at the same time, an effective lipid radical scavenger and lipid antioxidant. Here, we demonstrate that cardiolipin oxidation during apoptosis is realized not via a random cardiolipin peroxidation mechanism but rather proceeds as a result of peroxidase reaction in a tight cytochrome c/cardiolipin complex that restrains interactions of etoposide with radical intermediates generated in the course of the reaction. Using low-temperature and ambient-temperature electron paramagnetic resonance spectroscopy of H(2)O(2) induced protein-derived (tyrosyl) radicals and etoposide phenoxyl radicals, respectively, we established that cardiolipin peroxidation and etoposide oxidation by cytochrome c/cardiolipin complex takes place predominantly on protein-derived radicals of cytochrome c. We further show that etoposide can inhibit cytochrome c-catalyzed oxidation of cardiolipin competing with it as a peroxidase substrate. Peroxidase reaction of cytochrome c/cardiolipin complexes causes cross-linking and oligomerization of cytochrome c. With nonoxidizable tetraoleoyl-cardiolipin, the cross-linking occurs via dityrosine formation, whereas bifunctional lipid oxidation products generated from tetralinoleoyl cardiolipin participate in the production of high molecular weight protein aggregates. Protein aggregation is effectively inhibited by etoposide. The inhibition of cardiolipin peroxidation by etoposide, however, is realized at far higher concentrations than those at which it induces apoptotic cell death. Thus, oxidation of cardiolipin by the cytochrome c/cardiolipin peroxidase complex, which is essential for apoptosis, is not inhibited by proapoptotic concentrations of the drug. PMID- 16690783 TI - Mammalian tolloid alters subcellular localization, internalization, and signaling of alpha(1a)-adrenergic receptors. AB - In the present study, we identified the CUB5 domain of mammalian Tolloid (mTLD) as a novel protein binding to alpha(1A)-adrenergic receptor (AR) using the yeast two-hybrid system. Whereas CUB5 did not couple to either alpha(1B)-AR or alpha(1D)-AR. It was determined that amino acids 322 to 359 of alpha(1A)-AR were the major binding region for CUB5. The direct interaction between alpha(1A)-AR cytoplasmic tail and CUB5 was discovered by glutathione S-transferase pull-down assay. We confirmed the interaction of mTLD with alpha(1A)-AR in human embryonic kidney (HEK) 293 cells by immunoprecipitation, immunofluorescence, and fluorescence resonance energy transfer. Although mTLD did not affect the density and affinity of receptors in crudely prepared membranes from HEK293 cells stably expressing alpha(1A)-AR, it significantly altered the subcellular localization of the receptors. Moreover, mTLD reduced the level of cell surface alpha(1A)-ARs, delayed the initial rate of agonist-induced receptor internalization, and facilitated agonist-induced calcium transient. We have demonstrated that mTLD interacts with alpha(1A)-AR directly, alters the subcellular localization of receptor, and influences agonist-induced alpha(1A)-AR internalization and calcium signaling. PMID- 16690785 TI - Effects of hindlimb unweighting and aging on rat semimembranosus muscle and myosin. AB - We tested the hypothesis that lower specific force (force/cross-sectional area) generated by type II fibers from hindlimb-unweighted rats resulted from structural changes in myosin (i.e., a change in the ratio of myosin cross bridges in the weak- and strong-binding state during contraction). In addition, we determined whether those changes were age dependent. Permeabilized semimembranosus muscle fibers from young adult and aged rats, some of which were hindlimb unweighted for 3 wk, were studied for Ca(2+)-activated force generation and maximal unloaded shortening velocity. Fibers were also spin labeled specifically at myosin Cys707 to assess the structural distribution of myosin during maximal isometric contraction using electron paramagnetic resonance spectroscopy. Myosin heavy chain isoform (MHC) expression and the ratio of MHC to actin were evaluated in each fiber. Fibers from the unweighted rats generated 34% less specific force than fibers from weight-bearing rats (P < 0.001), independent of age. Electron paramagnetic resonance analyses showed that the fraction of myosin heads in the strong-binding structural state during contraction was 11% lower in fibers from the unweighted rats (P = 0.019), independent of age. More fibers from unweighted rats coexpressed MHC IIB-IIX compared with fibers from weight-bearing rats (P = 0.049). Unweighting induced a slowing of maximal unloaded shortening velocity and an increase in the ratio of MHC to actin in fibers from young rats only. These data indicate that altered myosin structural distribution during contraction and a preferential loss of actin contribute to unweighting-induced muscle weakness. Furthermore, the age of the rat has an influence on some parameters of changes in muscle contractility that are induced by unweighting. PMID- 16690784 TI - Activation of signaling pathways and regulatory mechanisms of mRNA translation following myocardial ischemia-reperfusion. AB - Protein expression in the heart is altered following periods of myocardial ischemia. The changes in protein expression are associated with increased cell size that can be maladaptive. There is little information regarding the regulation of protein expression through the process of mRNA translation during ischemia and reperfusion in the heart. Therefore, the purpose of this study was to identify changes in signaling pathways and downstream regulatory mechanisms of mRNA translation in an in vivo model of myocardial ischemia and reperfusion. Hearts were collected from rats whose left main coronary arteries had either been occluded for 25 min or reversibly occluded for 25 min and subsequently reperfused for 15 min. Following reperfusion, both the phosphoinositide 3-kinase and mitogen activated protein kinase pathways were activated, as evidenced by increased phosphorylation of Akt (PKB), extracellular signal-regulated kinase 1/2, and p38 mitogen-activated protein kinase. Activation of Akt stimulated signaling through the protein kinase mammalian target of rapamycin, as evidenced by increased phosphorylation of two of its effectors, the ribosomal protein S6 kinase and the eukaryotic initiation factor eIF4E binding protein 1. Ischemia and reperfusion also resulted in increased phosphorylation of eIF2 and eIF2B. These changes in protein phosphorylation suggest that control of mRNA translation following ischemia and reperfusion is modulated through a number of signaling pathways and regulatory mechanisms. PMID- 16690786 TI - Physiologic Interrelationships between Gibbs-Donnan Equilibrium, Osmolality of Body Fluid Compartments, and Plasma Water Sodium Concentration. AB - None. PMID- 16690787 TI - Lactic acid accumulation is an advantage/disadvantage during muscle activity. PMID- 16690788 TI - Continuous blood pressure-derived cardiac output monitoring--should we be thinking long term? PMID- 16690789 TI - Master equation for dysnatremia or intractable abracadabra. PMID- 16690790 TI - Reflexes of the lung and airways. PMID- 16690791 TI - Aging, muscle fiber type, and contractile function in sprint-trained athletes. AB - Biopsy samples were taken from the vastus lateralis of 18- to 84-yr-old male sprinters (n = 91). Fiber-type distribution, cross-sectional area, and myosin heavy chain (MHC) isoform content were identified using ATPase histochemistry and SDS-PAGE. Specific tension and maximum shortening velocity (V(o)) were determined in 144 single skinned fibers from younger (18-33 yr, n = 8) and older (53-77 yr, n = 9) runners. Force-time characteristics of the knee extensors were determined by using isometric contraction. The cross-sectional area of type I fibers was unchanged with age, whereas that of type II fibers was reduced (P < 0.001). With age there was an increased MHC I (P < 0.01) and reduced MHC IIx isoform content (P < 0.05) but no differences in MHC IIa. Specific tension of type I and IIa MHC fibers did not differ between younger and older subjects. V(o) of fibers expressing type I MHC was lower (P < 0.05) in older than in younger subjects, but there was no difference in V(o) of type IIa MHC fibers. An aging-related decline of maximal isometric force (P < 0.001) and normalized rate of force development (P < 0.05) of knee extensors was observed. Normalized rate of force development was positively associated with MHC II (P < 0.05). The sprint-trained athletes experienced the typical aging-related reduction in the size of fast fibers, a shift toward a slower MHC isoform profile, and a lower V(o) of type I MHC fibers, which played a role in the decline in explosive force production. However, the muscle characteristics were preserved at a high level in the oldest runners, underlining the favorable impact of sprint exercise on aging muscle. PMID- 16690792 TI - Sexual dimorphism of the intracellular heat shock protein 72 response. AB - The majority of previous work examining stress responses has been done in males. Recently, it has become clear that the impact of stressor exposure is modulated by sex. One stress response that may be affected by sex is the induction of intracellular heat shock protein (HSP) 72, which is a stress- responsive molecular chaperone that refolds denatured proteins and promotes cellular survival. The following study compared HSP72 in males and females and also examined whether the estrous cycle altered HSP72 induction in females. We hypothesized that females compared with males would have a constrained HSP72 response after an acute stressor and that the stress-induced HSP72 response in females would fluctuate with the estrous cycle. Male and female F344 rats were either left in their home cage or exposed to acute tail-shock stress (8 10/group). Immediately following stressor, trunk blood was collected and tissues were flash frozen. Vaginal smear and estrogen enzyme immunoassay were used to categorize the phase of estrous. Results show that female rats had a greater corticosterone response than males, that both males and females exhibit a stress induced release of progesterone, and that males and females had equal levels of stress-induced circulating norepinephrine. Sexual dimorphism of the HSP72 (ELISA) response existed in pituitary gland, mesenteric lymph nodes, and liver such that female rats had an attenuated HSP72 response compared with males after stress. The adrenal glands, spleen, and heart did not exhibit sexual dimorphism of the HSP72 response. The estrous cycle did not have a significant effect on basal or stress-induced HSP72 in females. PMID- 16690793 TI - Influence of acute plasma volume expansion on VO2 kinetics, VO2 peak, and performance during high-intensity cycle exercise. AB - The purpose of this study was to examine the influence of acute plasma volume expansion (APVE) on oxygen uptake (V(O2)) kinetics, V(O2peak), and time to exhaustion during severe-intensity exercise. Eight recreationally active men performed "step" cycle ergometer exercise tests at a work rate requiring 70% of the difference between the gas-exchange threshold and V(O2max) on three occasions: twice as a "control" (Con) and once after intravenous infusion of a plasma volume expander (Gelofusine; 7 ml/kg body mass). Pulmonary gas exchange was measured breath by breath. APVE resulted in a significant reduction in hemoglobin concentration (preinfusion: 16.0 +/- 1.0 vs. postinfusion: 14.7 +/- 0.8 g/dl; P < 0.001) and hematocrit (preinfusion: 44 +/- 2 vs. postinfusion: 41 +/- 3%; P < 0.01). Despite this reduction in arterial O(2) content, APVE had no effect on V(O2) kinetics (phase II time constant, Con: 33 +/- 15 vs. APVE: 34 +/- 12 s; P = 0.74), and actually resulted in an increased V(O2peak) (Con: 3.90 +/- 0.56 vs. APVE: 4.12 +/- 0.55 l/min; P = 0.006) and time to exhaustion (Con: 365 +/- 58 vs. APVE: 424 +/- 64 s; P = 0.04). The maximum O(2) pulse was also enhanced by the treatment (Con: 21.3 +/- 3.4 vs. APVE: 22.7 +/- 3.4 ml/beat; P = 0.04). In conclusion, APVE does not alter V(O2) kinetics but enhances V(O2peak) and exercise tolerance during high-intensity cycle exercise in young recreationally active subjects. PMID- 16690794 TI - Features of glossopharyngeal breathing in breath-hold divers. AB - One technique employed by competitive breath-hold divers to increase diving depth is to hyperinflate the lungs with glossopharyngeal breathing (GPB). Our aim was to assess the relationship between measured volume and pressure changes due to GPB. Seven healthy male breath-hold divers, age 33 (8) [mean (SD)] years were recruited. Subjects performed baseline body plethysmography (TLC(PRE)). Plethysmography and mouth relaxation pressure were recorded immediately following a maximal GPB maneuver at total lung capacity (TLC) (TLC(GPB)) and within 5 min after the final GPB maneuver (TLC(POST)). Mean TLC increased from TLC(PRE) to TLC(GPB) by 1.95 (0.66) liters and vital capacity (VC) by 1.92 (0.56) liters (P < 0.0001), with no change in residual volume. There was an increase in TLC(POST) compared with TLC(PRE) of 0.16 liters (0.14) (P < 0.02). Mean mouth relaxation pressure at TLC(GPB) was 65 (19) cmH(2)O and was highly correlated with the percent increase in TLC (R = 0.96). Breath-hold divers achieve substantial increases in measured lung volumes using GPB primarily from increasing VC. Approximately one-third of the additional air was accommodated by air compression. PMID- 16690795 TI - Cerebellar fastigial nuclei activity during blood pressure challenges. AB - The cerebellar fastigial nuclei (FN) assist in regulating compensatory responses to large blood pressure changes and show structural injury and functional impairment to cardiovascular challenges in syndromes with sleep-disordered breathing. The patterned time course of FN responses to elevation or lowering of blood pressure and location of responsive regions within the nuclei are unclear. We evaluated FN neural activity in six anesthetized rats using optical imaging procedures during elevation and lowering of arterial pressure by phenylephrine and nitroprusside, respectively. Hypertension diminished optical correlates of FN neural activity, while measures of activity increased to hypotension, with peak neural responses occurring 5-10 s later than peak blood pressure changes. Blood pressure responses were followed by heart rate changes, and peak respiratory rates developed even later, in close temporal proximity to FN activity patterns. Although overall topographical response trends were similar, regional patterns of altered neural activity appeared to both hypertension and hypotension. The extent of neural change was greater during recovery from hypertension than for hypotension at high-dose levels. Blood pressure levels saturated with increasing phenylephrine doses, while FN activity continued to decline. No saturation appeared in heart or respiratory rate trends. The findings suggest that the FN compensate for large blood pressure changes by sympathoexcitatory and inhibitory processes, which accompany late-developing somatic or respiratory adjustments. PMID- 16690796 TI - The Octopus vulgaris estrogen receptor is a constitutive transcriptional activator: evolutionary and functional implications. AB - Steroid hormones such as estrogens and androgens are important regulators of reproduction, physiology, and development in a variety of animal taxa, including vertebrates and mollusks. Steroid hormone receptors, which mediate the classic cellular responses to these hormones, were thought to be vertebrate specific, which left the molecular mechanisms of steroid action in invertebrates unresolved. Recently an estrogen receptor (ER) ortholog was isolated from the sea hare Aplysia californica, but the functional significance of the receptor was unclear because estrogens and other steroids are not known to be important in that species. Furthermore, the Aplysia ER was found to be a constitutive transcriptional activator, but it was unclear whether the estrogen independence of the ER was an Aplysia-specific novelty or a more ancient character general to the mollusks. Here we report on the isolation and functional characterization of the first ER ortholog from an invertebrate in which estrogens are produced and play an apparent role, the cephalopod Octopus vulgaris. We show that the Octopus ER is a strong constitutive transcriptional activator from canonical estrogen response elements. The receptor does not bind estradiol and is unresponsive to estrogens and other vertebrate steroid hormones. These characteristics are similar to those observed with the Aplysia ER and support the hypothesis that the evolving ER gained constitutive activity deep in the mollusk lineage. The apparent reproductive role of estrogens in Octopus and other mollusks is unlikely to be mediated by the ER and may take place through an ancient, non-ER-mediated pathway. PMID- 16690797 TI - Lectin-like oxidized low-density lipoprotein receptor-1-mediated autophagy in human granulosa cells as an alternative of programmed cell death. AB - The LOX-1 receptor, identified on endothelial cells, mediates the uptake of oxidized low-density lipoprotein (oxLDL). The oxLDL-dependent LOX-1 activation causes endothelial cell apoptosis. We here investigated the presence of LOX-1 in granulosa cells from patients under in vitro fertilization therapy. We were interested in the oxLDL-dependent LOX-1 receptor biology, in particular in the induction of apoptosis. In the human ovary, LOX-1 was localized in regressing antral follicles. In granulosa cell cultures, oxLDL-induced mRNA expression of LOX-1 in a time- and dose-dependent manner. The LOX-1 inhibitors (anti-LOX-1 antibody and kappa-carrageenan) abrogated the up-regulation of LOX-1. The oxLDL (100 microg/ml) treatment caused the autophagy form of programmed cell death: 1) reorganization of the actin cytoskeleton at the 6-h time point; 2) uptake of YO PRO, a marker for the early step of programmed cell death, before propidium iodide staining to signify necrosis; 3) absence of apoptotic bodies and cleaved caspase-3; 4) abundant vacuole formation at the ultrastructural level; and 5) decrease of the autophagosome marker protein MAP LC3-I at the 6-h time point indicative of autophagosome formation. We conclude that follicular atresia is not under the exclusive control of apoptosis. The LOX-1-dependent autophagy represents an alternate form of programmed cell death. Obese women with high blood levels of oxLDL may display an increased rate of autophagic granulosa cell death. PMID- 16690798 TI - Endocrine disruption for endocrinologists (and others). PMID- 16690799 TI - Energized, polarized, and actively respiring mitochondria are required for acute Leydig cell steroidogenesis. AB - The first and rate-limiting step in the biosynthesis of steroid hormones is the transfer of cholesterol into mitochondria, which is facilitated by the steroidogenic acute regulatory (StAR) protein. Recent study of Leydig cell function has focused on the mechanisms regulating steroidogenesis; however, few investigations have examined the importance of mitochondria in this process. The purpose of this investigation was to determine which aspects of mitochondrial function are necessary for acute cAMP-stimulated Leydig cell steroidogenesis. MA 10 cells were treated with 8-bromoadenosine 3',5'-cyclic monophosphate (cAMP) and different site-specific agents that disrupt mitochondrial function, and the effects on acute cAMP-stimulated progesterone synthesis, StAR mRNA and protein, mitochondrial membrane potential (Deltapsim), and ATP synthesis were determined. cAMP treatment of MA-10 cells resulted in significant increases in both cellular respiration and Deltapsim. Dissipating Deltapsim with carbonyl cyanide m chlorophenyl hydrazone resulted in a profound reduction in progesterone synthesis, even in the presence of newly synthesized StAR protein. Preventing electron transport in mitochondria with antimycin A significantly reduced cellular ATP, potently inhibited steroidogenesis, and reduced StAR protein levels. Inhibiting mitochondrial ATP synthesis with oligomycin reduced cellular ATP, inhibited progesterone synthesis and StAR protein, but had no effect on Deltapsim. Disruption of intramitochondrial pH with nigericin significantly reduced progesterone production and StAR protein but had minimal effects on Deltapsim. 22(R)-hydroxycholesterol-stimulated progesterone synthesis was not inhibited by any of the mitochondrial reagents, indicating that neither P450 side chain cleavage nor 3beta-hydroxysteroid dehydrogenase activity was inhibited. These results indicate that Deltapsim, mitochondrial ATP synthesis, and mitochondrial pH are all required for acute steroid biosynthesis. These results suggest that mitochondria must be energized, polarized, and actively respiring to support Leydig cell steroidogenesis, and alterations in the state of mitochondria may be involved in regulating steroid biosynthesis. PMID- 16690800 TI - The aryl hydrocarbon receptor pathway and sexual differentiation of neuroendocrine functions. AB - Historically, much of the research on health effects of environmental pollutants focused on ascertaining whether compounds were carcinogenic. More recent findings show that environmental contaminants also exert insidious effects by disrupting hormone action. Of particular concern are findings that developmental exposure to dioxins, chemicals that act through the aryl hydrocarbon receptor pathway, permanently alters sexually differentiated neural functions in animal models. In this review, we focus on mechanisms through which dioxins disrupt neuroendocrine development as exemplified by effects on a brain region critical for ovulation in rodents. We also provide evidence that dysregulation of GABAergic neural development may be a general mechanism underlying a broad spectrum of effects seen after perinatal dioxin exposure. PMID- 16690801 TI - Environmental obesogens: organotins and endocrine disruption via nuclear receptor signaling. AB - Over the last two decades, the incidence of obesity and associated metabolic syndrome diseases has risen dramatically, becoming a global health crisis. Increased caloric intake and decreased physical activity are believed to represent the root causes of this dramatic rise. However, recent findings highlight the possible involvement of environmental obesogens, xenobiotic chemicals that can disrupt the normal developmental and homeostatic controls over adipogenesis and energy balance. Environmental estrogens, i.e. chemicals with estrogenic potential, have been reported to perturb adipogenic mechanisms using in vitro model systems, but other classes of endocrine-disrupting chemicals are now coming under scrutiny as well. Organotins represent one class of widespread persistent organic pollutants with potent endocrine-disrupting properties in both invertebrates and vertebrates. New data identify tributyltin chloride and triphenyltin chloride as nanomolar agonist ligands for retinoid X receptor (RXR alpha, RXR beta, and RXR gamma) and peroxisome proliferator-activated receptor gamma, nuclear receptors that play pivotal roles in lipid homeostasis and adipogenesis. The environmental obesogen hypothesis predicts that inappropriate receptor activation by organotins will lead directly to adipocyte differentiation and a predisposition to obesity and/or will sensitize exposed individuals to obesity and related metabolic disorders under the influence of the typical high calorie, high-fat Western diet. The linking of organotin exposure to adipocyte differentiation and obesity opens an important new area of research into potential environmental influences on human health and disease. PMID- 16690802 TI - Endocrine-disrupting chemicals use distinct mechanisms of action to modulate endocrine system function. AB - The term endocrine-disrupting chemicals is used to define a structurally diverse class of synthetic and natural compounds that possess the ability to alter various components of the endocrine system and potentially induce adverse health effects in exposed individuals and populations. Research on these compounds has revealed that they use a variety of both nuclear receptor-mediated and non receptor-mediated mechanisms to modulate different components of the endocrine system. This review will describe in vitro and in vivo studies that highlight the spectrum of unique mechanisms of action and biological effects of four endocrine disrupting chemicals--diethylstilbestrol, genistein, di(n-butyl)phthalate, and methoxyacetic acid--to illustrate the diverse and complex nature of this class of compounds. PMID- 16690803 TI - Epigenetic transgenerational actions of endocrine disruptors. AB - Endocrine disruptors have recently been shown to promote an epigenetic transgenerational phenotype involving a number of disease states (e.g. male infertility). The anti-androgenic fungicide vinclozolin was found to act transiently at the time of embryonic sex determination to promote in the F1 generation a spermatogenic cell defect and subfertility in the male. When the animals were allowed to age up to 1 yr, a number of other disease states developed. This phenotype was transferred through the male germ line to all subsequent generations analyzed (F1-F4). The ability of an environmental factor (i.e. endocrine disruptor) to promote an epigenetic transgenerational phenotype impacts the potential hazards of environmental toxins, mechanisms of disease etiology, and evolutionary biology. The biological importance of the epigenetic actions of environmental agents is reviewed in the context of the primordial germ cell and development of epigenetic transgenerational phenotypes. PMID- 16690804 TI - Estrogen regulation of the glucuronidation enzyme UGT2B15 in estrogen receptor positive breast cancer cells. AB - Estrogens and androgens influence many properties of breast cancer cells; hence, regulation of local estrogen and androgen levels by enzymes involved in steroid hormone biosynthesis and metabolism would impact signaling by these hormones in breast cancer cells. In this study, we show that the UDP-glucuronosyltransferase (UGT) enzyme UGT2B15, a member of the UGT family of phase II enzymes involved in the glucuronidation of steroids and xenobiotics, is a novel, estrogen-regulated gene in estrogen receptor (ER)-positive human breast cancer cells (MCF-7, BT474, T47D, and ZR-75). UGT2B15 is the only UGT2B enzyme up-regulated by estrogen, and marked estradiol stimulation of UGT2B15 mRNA levels is observed, in a time- and dose-dependent manner. UGT2B15 stimulation by estradiol is blocked by the antiestrogen ICI182,780, but not by the translational inhibitor cycloheximide, indicating that UGT2B15 is likely a primary transcriptional response mediated through the ER. UGT2B15 up-regulation is also evoked by other estrogens (propylpyrazoletriol, genistein) and by the androgen 5alpha-dihydrotestosterone working through the ER, but not by other steroid hormone receptor ligands. Western blot and immunocytochemical analyses with several UGT2B-specific antibodies we have designed and steroid glucuronidation assays indicate a large increase in both cellular UGT2B15 protein and enzyme activity after estrogen treatment. Due to the important role of UGT enzymes in forming conjugates between steroids and glucuronic acid, thereby inactivating them and targeting them for removal, the estrogen-induced up-regulation of UGT2B15 might have a significant moderating effect on estrogen and androgen concentrations, thereby reducing their signaling in breast cancer cells. PMID- 16690805 TI - Regulation of membrane-associated prostaglandin E2 synthase 1 in pregnant sheep intrauterine tissues by glucocorticoid and estradiol. AB - This study was designed to determine the regulatory effect of glucocorticoid and estradiol on expression of ovine intrauterine membrane-associated prostaglandin E(2) synthase 1 (mPTGES1) in late gestation and at labor. For gestational and labor groups, 16 pregnant ewes from 95-147 d gestational age (dGA) and four pregnant ewes at spontaneous term labor were used. The fetal glucocorticoid group, 14 pregnant ewes at 123-125 dGA with fetuses, was divided into the following groups: after sham adrenalectomy (n = 5), adrenalectomy (n = 4), and adrenalectomy with fetal cortisol replacement to late gestation levels (n = 5). For the maternal glucocorticoid group, nine pregnant ewes were treated with saline (n = 4) and three courses of maternal dexamethasone (n = 5). For the estradiol group, 10 pregnant ewes at 119-121 dGA were treated with sesame oil (n = 5) or estradiol (n = 5) to produce labor levels of estradiol in maternal plasma. Endometrial, myometrial, and placental mRNA and proteins were analyzed by Northern and Western blot and immunocytochemistry for mPTGES1. Data were analyzed by Student's t test and ANOVA. There was a significant increase of placental mPTGES1 in late gestation. Glucocorticoids, given to the mother or fetus, significantly stimulated mPTGES1 in placenta. mPTGES1 was elevated only in the endometrium during spontaneous term labor and after estradiol treatment. The mPTGES1 was localized in the myometrial smooth muscle cells, endometrial stromal cells, and placental trophoblast cells. Our study suggested that increased expression of placental mPTGES1 throughout late gestation might result from the increased fetal and maternal circulating glucocorticoids, whereas elevated maternal plasma estradiol concentration might be responsible for the induced mPTGES1 expression in the endometrium during labor. PMID- 16690806 TI - FOXO1A differentially regulates genes of decidualization. AB - The forkhead box O1A (FOXO1A) has been identified as one gene that is up regulated early in the decidualization process. To further investigate the role of FOXO1A during this process, six genes, IGFBP1, PRL, TIMP3, LAMB1, CNR1, and DCN, shown to be up-regulated during decidualization, were chosen as potential targets of FOXO1A action. Treatment of human endometrial stromal cells with hormones (estradiol and medroxyprogesterone acetate) plus dibutyryl cAMP (H+dbcAMP) for 48 h increased expression of IGFBP1, PRL, TIMP3, CNR1, and DCN but not LAMB1, as measured by real-time PCR. Silencing of FOXO1A using small interfering RNA oligonucleotides decreased IGFBP1 and DCN levels and increased CNR1, TIMP3, and PRL levels. LAMB1 was not affected. When FOXO1A was overexpressed in human endometrial stromal cells, expression of IGFBP1, DCN, and PRL increased, whereas levels of TIMP3 and CNR1 decreased. Addition of H+dbcAMP caused an increased expression of IGFBP1, PRL, and DCN beyond that of FOXO1A alone. TIMP3 and CNR1 levels decreased even further in response to H+dbcAMP compared with FOXO1A alone. LAMB1, which was unresponsive to FOXO1A, decreased when H+dbcAMP was added. Overexpressing FOXO1A also caused a change in cell shape, in that the stromal fibroblasts acquired a rounded, epithelioid appearance. Finally, reporter studies showed that cotransfection of FOXO1A significantly increased PRL promoter activity but not TIMP3 promoter activity. Addition of H+dbcAMP resulted in a significant increase in PRL promoter activity and a significant decrease in TIMP3 promoter activity. In summary, this study demonstrates the versatile nature of FOXO1A in the regulation of a number of decidualization-specific genes. PMID- 16690807 TI - Cortisol stimulates cell cycle activity in the cardiomyocyte of the sheep fetus. AB - The role of cortisol in regulating cardiac myocyte growth in the near-term fetal sheep is unknown. We hypothesized that cortisol would suppress cardiomyocyte proliferation and stimulate cardiomyocyte binucleation and enlargement, signs of terminal differentiation. Cardiomyocyte dimensions and percent binucleation were determined in isolated cardiac myocytes from seven cortisol-treated and seven control fetuses; percentage of myocytes positive for Ki-67 was determined in an additional four cortisol-treated and four control hearts. Cortisol was infused into the circumflex coronary artery at subpressor rates (0.5 microg/kg.min, 7 d). Cortisol infusion had no hemodynamic effects, compared with controls or pretreatment conditions. Cortisol treatment increased heart weight (44.0 +/- 8.7 g vs. control, 34.9 +/- 9.1 g, P < 0.05). Heart to body weight ratio was greater in treated hearts, compared with controls (10.3 +/- 1.9 vs. 7.7 +/- 0.9 g/kg, P < 0.01). Ventricular myocyte length, width, and percent binucleation were not different between groups. The proportion of treated myocytes in the cell cycle staining for Ki-67 was higher in the left ventricle (5.5 +/- 0.1 vs. 2.7 +/- 0.4%, P < 0.005) and right ventricle (4.4 +/- 0.4 vs. 3.7 +/- 0.7%, P < 0.05), compared with controls. Wet weight to dry weight ratios from cortisol-treated and control hearts were not different. In conclusion, whereas cortisol infused into the fetal sheep heart has no effect on cardiomyocyte size or maturational state, it stimulates entry of cardiomyocytes in the cell cycle. Thus, increases in fetal heart mass associated with subpressor doses of cortisol are due to cardiomyocyte proliferation and not hypertrophic growth. PMID- 16690808 TI - Pyridoxal phosphate inhibits pituitary cell proliferation and hormone secretion. AB - Pyridoxal phosphate (PLP), a bioactive form of pyridoxine, dose-dependently (10 1000 microm) inhibited cell proliferation in rat pituitary MMQ and GH3 cells and in mouse AtT-20 cells. After 4 d, MMQ cell numbers were reduced by up to 81%, GH3 cell numbers were reduced by up to 64% (P < 0.05), and AtT-20 cell numbers were reduced by up to 90%. Cell proliferation rates recovered and dose-dependently reverted to control levels after PLP withdrawal. After 4 d, PLP (400 and 1000 microm) decreased [3H]thymidine incorporation by up to 71% (P < 0.05). PLP (400 1000 microm) reduced GH3 cell GH and prolactin secretion and AtT-20 cell ACTH secretion (adjusted for cell number) by approximately 70% after 2 d. The 100 microm PLP also inhibited prolactin secretion (65%, P < 0.05) in primary rat pituitary cells treated for 2 d. PLP decreased the percentage of AtT-20 and GH3 cells in S phase and increased those in G0-G1 phase. Furthermore, PLP induced AtT 20 and GH3 cell apoptosis (28 vs. 6, P < 0.05; 26 vs. 3, P < 0.05, respectively) and dose-dependently reduced content of the antiapoptosis gene Bcl-2. These results indicate that pharmacological doses of PLP inhibit pituitary cell proliferation and hormone secretion, in part mediated through PLP-induced cell cycle arrest and apoptosis. Pyridoxine may therefore be appropriate for testing as a relatively safe drug for adjuvant treatment of hormone-secreting pituitary adenomas. PMID- 16690809 TI - Adverse effects of the model environmental estrogen diethylstilbestrol are transmitted to subsequent generations. AB - The synthetic estrogen diethylstilbestrol (DES) is a potent perinatal endocrine disruptor. In humans and experimental animals, exposure to DES during critical periods of reproductive tract differentiation permanently alters estrogen target tissues and results in long-term abnormalities such as uterine neoplasia that are not manifested until later in life. Using the developmentally exposed DES mouse, multiple mechanisms have been identified that play a role in its carcinogenic and toxic effects. Analysis of the DES murine uterus reveals altered gene expression pathways that include an estrogen-regulated component. Thus, perinatal DES exposure, especially at low doses, offers the opportunity to study effects caused by weaker environmental estrogens and provides an example of the emerging scientific field termed the developmental origin of adult disease. As a model endocrine disruptor, it is of particular interest that even low doses of DES increase uterine tumor incidence. Additional studies have verified that DES is not unique; when other environmental estrogens are tested at equal estrogenic doses, developmental exposure results in increased incidence of uterine neoplasia similar to that caused by DES. Interestingly, our data suggest that this increased susceptibility for tumors is passed on from the maternal lineage to subsequent generations of male and female descendants; the mechanisms involved in these transgenerational events include genetic and epigenetic events. Together, our data point out the unique sensitivity of the developing organism to endocrine disrupting chemicals, the occurrence of long-term effects after developmental exposure, and the possibility for adverse effects to be transmitted to subsequent generations. PMID- 16690810 TI - Large effects from small exposures. III. Endocrine mechanisms mediating effects of bisphenol A at levels of human exposure. AB - Over 6 billion pounds per year of the estrogenic monomer bisphenol A (BPA) are used to manufacture polycarbonate plastic products, in resins lining metal cans, in dental sealants, and in blends with other types of plastic products. The ester bond linking BPA molecules in polycarbonate and resins undergoes hydrolysis, resulting in the release of free BPA into food, beverages, and the environment, and numerous monitoring studies now show almost ubiquitous human exposure to biologically active levels of this chemical. BPA exerts estrogenic effects through the classical nuclear estrogen receptors, and BPA acts as a selective estrogen receptor modulator. However, BPA also initiates rapid responses via estrogen receptors presumably associated with the plasma membrane. Similar to estradiol, BPA causes changes in some cell functions at concentrations between 1 pM and 1 nM, and the mean and median range of unconjugated BPA measured by multiple techniques in human pregnant maternal, fetal, and adult blood and other tissues exceeds these levels. In contrast to these published findings, BPA manufacturers persist in describing BPA as a weak estrogen and insist there is little concern with human exposure levels. Our concern with human exposure to BPA derives from 1) identification of molecular mechanisms mediating effects in human and animal tissues at very low doses, 2) in vivo effects in experimental animals caused by low doses within the range of human exposure, and 3) widespread human exposure to levels of BPA that cause adverse effects in animals. PMID- 16690811 TI - Endocrine-disrupting compounds and mammary gland development: early exposure and later life consequences. AB - Breast cancer is the most common non-skin cancer among women in this country. Breast cancer risk is significantly influenced by genetics, but over 70% of the women that are diagnosed have noninherited or sporadic cancer. The risk of breast cancer is thought to be modified by lifestyle and environment. Exposures to certain chemicals and hormone-mimicking or endocrine-disrupting compounds (EDCs) are suspected of contributing to increased breast cancer incidence as well as precocious puberty in the United States. Studies of EDC effects in rodents indicate that multiple toxicants can alter mammary gland development, with or without changing other markers of puberty. EDCs can cause transient and persistent effects on mammary gland development depending on dose, exposure parameters, and whether exposure was during critical periods of gland growth or differentiation. Adverse effects from these abnormal developmental patterns include the presence of carcinogen-sensitive structures in greater numbers or for longer periods in the gland and inhibited functional differentiation leading to malnutrition or increased mortality of their offspring. Developmental toxicants of the mammary gland could lead to an increase in the incidence of mammary tumors if they alter circulating or tissue-localized hormone levels, gland receptor expression patterns, hormone transport, or metabolism that results in altered response to endogenous hormones or growth factors. Environmental disruptors of rodent mammary gland development must be identified for informed decisions in epidemiological studies aimed at identification of environmental factors contributing to breast cancer risk, altered breast development during puberty, or inability to produce sufficient breast milk. PMID- 16690812 TI - Epigenetics, evolution, endocrine disruption, health, and disease. AB - Endocrine-disrupting chemicals (EDCs) in the environment have been linked to human health and disease. This is particularly evident in compounds that mimic the effects of estrogens. Exposure to EDCs early in life can increase risk levels of compromised physical and mental health. Epigenetic mechanisms have been implicated in this process. Transgenerational consequences of EDC exposure is also discussed in both a proximate (mechanism) and ultimate (evolution) context as well as recent work suggesting how such transmission might become incorporated into the genome and subject to selection. We suggest a perspective for exploring and ultimately coming to understand diseases that may have environmental or endocrine origins. PMID- 16690813 TI - Point-of-care test identifies diabetic ketoacidosis at triage. AB - BACKGROUND: Diabetic ketoacidosis (DKA) is a common, life-threatening complication of diabetes. The diagnosis of DKA relies on signs and symptoms, plus laboratory findings of blood glucose (BG) of > 250 mg/dL, an anion gap (AG) of > or = 15 mmol/L, and carbon dioxide (CO2) of < or = 18 mmol/L when other causes of acidosis are excluded. OBJECTIVES: To compare the results of a point-of-care test for the ketone beta-hydroxybutyrate (beta-OHB) with standard measures for accuracy in predicting DKA. METHODS: After providing informed consent, 160 patients who presented with BG of > 250 mg/dL underwent testing for beta-OHB with the Precision Xtra meter (Abbott Laboratories, North Chicago, IL) at triage in a large urban hospital emergency department. The diagnosis of DKA was made by clinicians by using standard clinical criteria without knowledge of the beta-OHB test. RESULTS: A diagnosis of DKA was made in 57 of 160 subjects. The beta-OHB values correlated strongly with AG (r = 0.66, p < 0.001) and with CO2 (r = -0.69, p < 0.001), as well as with glucose (r = 0.31, p < 0.001). Cross-classification of DKA vs. beta-OHB yielded sensitivity of 98% (95% CI = 91% to 100%), specificity of 85% (95% CI = 78% to 91%), with a positive likelihood ratio of 6.7 (95% CI = 4.22 to 10.78), and negative likelihood ratio of 0.021 (95% CI = 0.003 to 0.144) at the manufacturer-suggested beta-OHB level of 1.5. CONCLUSIONS: The point-of-care test for beta-OHB was as sensitive as more established indicators of DKA. It is more useful than glucose alone for the diagnosis of DKA and offers immediate diagnosis of patients at triage. PMID- 16690814 TI - Lumbar puncture ordering and results in the pediatric population: a promising data source for surveillance systems. AB - BACKGROUND: The Centers for Disease Control and Prevention is incorporating laboratory data into real-time surveillance systems. When normal patterns of laboratory test orders and results are modeled, aberrations can be detected. Because many test orders are available electronically well before results, atypical patterns of test ordering may signal outbreaks. OBJECTIVES: The authors sought to characterize baseline patterns in the ordering and early results of lumbar punctures, motivated by the possibility of using these data for real-time surveillance for early detection of meningitis or encephalitis outbreaks. METHODS: Retrospective cohorts of pediatric emergency department patients at a single hospital (1993-2003) and from the National Hospital and Ambulatory Medical Care Survey (1992-2000) were used for analysis. RESULTS: Test ordering exhibits seasonal patterns, with monthly peaks in January and August (p < 0.0001). For the hospital cohort, the rate of cerebrospinal fluid pleocytosis exhibits seasonal patterns (p < 0.0001), with a peak from August to October. This is strongly associated with the rate and pattern of clinical neurologic disease (p < 0.0001). A long-term secular decline in daily test ordering is evident, dropping from 5.3 to 2.9 in the hospital sample, and from 371.8 to 185.3 in the national sample (p < 0.001). The long-term rate of pleocytosis has declined (p < 0.0001), though the yield of testing for pleocytosis has improved (p = 0.0104). CONCLUSIONS: Laboratory test patterns correspond with those of clinical disease and are a promising source of surveillance data. Using such data for real-time monitoring requires specific adjustments for patient age, periodicities, and secular trends. PMID- 16690815 TI - A new genus of African monkey, Rungwecebus: morphology, ecology, and molecular phylogenetics. AB - A new species of African monkey, Lophocebus kipunji, was described in 2005 based on observations from two sites in Tanzania. We have since obtained a specimen killed by a farmer on Mount Rungwe, the type locality. Detailed molecular phylogenetic analyses of this specimen demonstrate that the genus Lophocebus is diphyletic. We provide a description of a new genus of African monkey and of the only preserved specimen of this primate. We also present information on the animal's ecology and conservation. PMID- 16690816 TI - AXR4 is required for localization of the auxin influx facilitator AUX1. AB - The AUX1 and PIN auxin influx and efflux facilitators are key regulators of root growth and development. For root gravitropism to occur, AUX1 and PIN2 must transport auxin via the lateral root cap to elongating epidermal cells. Genetic studies suggest that AXR4 functions in the same pathway as AUX1. Here we show that AXR4 is a previously unidentified accessory protein of the endoplasmic reticulum (ER) that regulates localization of AUX1 but not of PIN proteins. Loss of AXR4 resulted in abnormal accumulation of AUX1 in the ER of epidermal cells, indicating that the axr4 agravitropic phenotype is caused by defective AUX1 trafficking in the root epidermis. PMID- 16690817 TI - Human genetics. Finding criminals through DNA of their relatives. PMID- 16690818 TI - Geochemistry. The pathway of carbon in nature. PMID- 16690819 TI - Biomarker evidence for a major preservation pathway of sedimentary organic carbon. AB - Hydrogenation processes leading from biomolecules to fossil biomarkers in anoxic sediments are crucial for the preservation of organic matter. However, these processes are still poorly understood. The present identification of several reduced carotenoids in recent sediments attests that these processes operate at the earliest stages of diagenesis without structural or stereochemical specificity, implying a nonbiological reduction pathway. Sulfur species (e.g., H2S) are the hydrogen donors involved in such reduction, as demonstrated with laboratory experiments. These reactions allow the preservation of abundant organic carbon in the rock record. PMID- 16690820 TI - Nanoassembly of a fractal polymer: a molecular "Sierpinski hexagonal gasket". AB - Mathematics and art converge in the fractal forms that also abound in nature. We used molecular self-assembly to create a synthetic, nanometer-scale, Sierpinski hexagonal gasket. This nondendritic, perfectly self-similar fractal macromolecule is composed of bis-terpyridine building blocks that are bound together by coordination to 36 Ru and 6 Fe ions to form a nearly planar array of increasingly larger hexagons around a hollow center. PMID- 16690821 TI - More silliness on the Hill. PMID- 16690822 TI - NASA budget. Crisis deepens as scientists fail to rejigger space research. PMID- 16690823 TI - Global change. No doubt about it, the world is warming. PMID- 16690824 TI - Intellectual property. Decision on NFkappaB patent could have broad implications for biotech. PMID- 16690826 TI - Condensed matter physics. Solid hydrogen not so super after all. PMID- 16690825 TI - Scholarly publishing. Bill would require free public access to research papers. PMID- 16690827 TI - U.S. science policy. Senate panel chair asks why NSF funds social sciences. PMID- 16690828 TI - Canada. Research budgets are tight pending science policy review. PMID- 16690829 TI - Scientific publishing. A call to improve South Africa's journals. PMID- 16690830 TI - Global health. Polio eradication: is it time to give up? PMID- 16690831 TI - Scientific publishing. A cure for the common trial. PMID- 16690832 TI - Cognitive Neuroscience Society meeting. Probing the social brain. PMID- 16690833 TI - Astronomy. A Hawaiian upstart prepares to monitor the starry heavens. PMID- 16690840 TI - Multiple outbreaks and flu containment plans. PMID- 16690841 TI - Migratory birds and avian flu. PMID- 16690842 TI - Species diversity and ecosystem functioning. PMID- 16690843 TI - Increase in foreign grad students. PMID- 16690844 TI - Comment on "Evidence for positive epistasis in HIV-1". AB - Bonhoeffer et al. (Reports, 26 November 2004, p. 1547) presented evidence for positive epistasis in a clinical data set of HIV-1 mutants and corresponding fitness values. We demonstrate that biases in the original and simulated data sets may lead to erroneous evidence for epistasis. More rigorous statistical tests must be used to account for such biases before one can infer epistasis. PMID- 16690845 TI - Public health. Progress toward rotavirus vaccines. PMID- 16690846 TI - Public health. Is polio eradication realistic? PMID- 16690847 TI - Public health. Who should get influenza vaccine when not all can? PMID- 16690848 TI - Ecology. Photosymbiosis and the evolution of modern coral reefs. PMID- 16690849 TI - Plant science. Auxin transport, but in which direction? PMID- 16690850 TI - Materials science. Toward devices powered by biomolecular motors. PMID- 16690851 TI - Neuroscience. Regulating energy balance: the substrate strikes back. PMID- 16690852 TI - Materials science. Collective defect behavior under stress. PMID- 16690853 TI - Geophysics. Ships' logs and archeomagnetism. PMID- 16690855 TI - Did DNA come from viruses? PMID- 16690856 TI - Viruses: making friends with old foes. AB - The study of viruses has traditionally focused on their roles as infectious agents and as tools for understanding cell biology. Viruses are now finding a new expanded role as nanoplatforms with applications in materials science and medicine. Viruses form highly symmetrical monodisperse architectures and are ideal templates for engineering multifunctionality, including multivalent display of surface ligands and encapsulation of inorganic and organic materials. These developments assure that viruses will find applications as versatile nanoscale materials. PMID- 16690857 TI - Aggresomes and autophagy generate sites for virus replication. AB - The replication of many viruses is associated with specific intracellular compartments called virus factories or virioplasm. These are thought to provide a physical scaffold to concentrate viral components and thereby increase the efficiency of replication. The formation of virus replication sites often results in rearrangement of cellular membranes and reorganization of the cytoskeleton. Similar rearrangements are seen in cells in response to protein aggregation, where aggresomes and autophagosomes are produced to facilitate protein degradation. Here I review the evidence that some viruses induce aggresomes and autophagosomes to generate sites of replication. PMID- 16690858 TI - Type 1 interferons and the virus-host relationship: a lesson in detente . AB - The interface between an infectious agent and its host represents the ultimate battleground for survival: The microbe must secure a niche for replication, whereas the host must limit the pathogen's advance. Among the host's arsenal of antimicrobial factors, the type 1 interferons (IFNs) induce potent defense mechanisms against viruses and are key in the host-virus standoff. Viruses have evolved multiple tricks to avoid the immediate antiviral effects of IFNs and, in turn, hosts have adapted use of this innate cytokine system to galvanize multiple additional layers of immune defense. The plasticity that exists in these interactions provides us with a lesson in detente. PMID- 16690859 TI - Formation and subdivision of deformation structures during plastic deformation. AB - During plastic deformation of metals and alloys, dislocations arrange in ordered patterns. How and when these self-organization processes take place have remained elusive, because in situ observations have not been feasible. We present an x-ray diffraction method that provided data on the dynamics of individual, deeply embedded dislocation structures. During tensile deformation of pure copper, dislocation-free regions were identified. They showed an unexpected intermittent dynamics, for example, appearing and disappearing with proceeding deformation and even displaying transient splitting behavior. Insight into these processes is relevant for an understanding of the strength and work-hardening of deformed materials. PMID- 16690860 TI - Simultaneous negative phase and group velocity of light in a metamaterial. AB - We investigated the propagation of femtosecond laser pulses through a metamaterial that has a negative index of refraction for wavelengths around 1.5 micrometers. From the interference fringes of a Michelson interferometer with and without the sample, we directly inferred the phase time delay. From the pulse envelope shift, we determined the group time delay. In a spectral region, phase and group velocity are negative simultaneously. This means that both the carrier wave and the pulse envelope peak of the output pulse appear at the rear side of the sample before their input pulse counterparts have entered the front side of the sample. PMID- 16690861 TI - Observation of backward pulse propagation through a medium with a negative group velocity. AB - The nature of pulse propagation through a material with a negative value of the group velocity has been mysterious, as simple models seem to predict that pulses will propagate "backward" through such a material. Using an erbium-doped optical fiber and measuring the time evolution of the pulse intensity at many points within the fiber, we demonstrate that the peak of the pulse does propagate backward inside the fiber, even though the energy flow is always in the forward direction. PMID- 16690862 TI - Statistical independence of escalatory ecological trends in Phanerozoic marine invertebrates. AB - Ecological interactions, such as predation and bioturbation, are thought to be fundamental determinants of macroevolutionary trends. A data set containing global occurrences of Phanerozoic fossils of benthic marine invertebrates shows escalatory trends in the relative frequency of ecological groups, such as carnivores and noncarnivorous infaunal or mobile organisms. Associations between these trends are either statistically insignificant or interpretable as preservational effects. Thus, there is no evidence that escalation drives macroecological trends at global and million-year time scales. We also find that taxonomic richness and occurrence data are cross-correlated, which justifies the traditional use of one as a proxy of the other. PMID- 16690863 TI - Fall in Earth's magnetic field is erratic. AB - Earth's magnetic field has decayed by about 5% per century since measurements began in 1840. Directional measurements predate those of intensity by more than 250 years, and we combined the global model of directions with paleomagnetic intensity measurements to estimate the fall in strength for this earlier period (1590 to 1840 A.D.). We found that magnetic field strength was nearly constant throughout this time, in contrast to the later period. Extrapolating to the core surface showed that the fall in strength originated in patches of reverse magnetic flux in the Southern Hemisphere. These patches were detectable by directional data alone; the pre-1840 model showed little or no evidence of them, supporting the conclusion of a steady dipole up to 1840. PMID- 16690864 TI - Impaired control of IRES-mediated translation in X-linked dyskeratosis congenita. AB - The DKC1 gene encodes a pseudouridine synthase that modifies ribosomal RNA (rRNA). DKC1 is mutated in people with X-linked dyskeratosis congenita (X-DC), a disease characterized by bone marrow failure, skin abnormalities, and increased susceptibility to cancer. How alterations in ribosome modification might lead to cancer and other features of the disease remains unknown. Using an unbiased proteomics strategy, we discovered a specific defect in IRES (internal ribosome entry site)-dependent translation in Dkc1(m) mice and in cells from X-DC patients. This defect results in impaired translation of messenger RNAs containing IRES elements, including those encoding the tumor suppressor p27(Kip1) and the antiapoptotic factors Bcl-xL and XIAP (X-linked Inhibitor of Apoptosis Protein). Moreover, Dkc1(m) ribosomes were unable to direct translation from IRES elements present in viral messenger RNAs. These findings reveal a potential mechanism by which defective ribosome activity leads to disease and cancer. PMID- 16690865 TI - RNA recognition and cleavage by a splicing endonuclease. AB - The RNA splicing endonuclease cleaves two phosphodiester bonds within folded precursor RNAs during intron removal, producing the functional RNAs required for protein synthesis. Here we describe at a resolution of 2.85 angstroms the structure of a splicing endonuclease from Archaeglobus fulgidus bound with a bulge-helix-bulge RNA containing a noncleaved and a cleaved splice site. The endonuclease dimer cooperatively recognized a flipped-out bulge base and stabilizes sharply bent bulge backbones that are poised for an in-line RNA cleavage reaction. Cooperativity arises because an arginine pair from one catalytic domain sandwiches a nucleobase within the bulge cleaved by the other catalytic domain. PMID- 16690866 TI - Molecular sorting by electrical steering of microtubules in kinesin-coated channels. AB - Integration of biomolecular motors in nanoengineered structures raises the intriguing possibility of manipulating materials on nanometer scales. We have managed to integrate kinesin motor proteins in closed submicron channels and to realize active electrical control of the direction of individual kinesin propelled microtubule filaments at Y junctions. Using this technique, we demonstrate molecular sorting of differently labeled microtubules. We attribute the steering of microtubules to electric field-induced bending of the leading tip. From measurements of the orientation-dependent electrophoretic motion of individual, freely suspended microtubules, we estimate the net applied force on the tip to be in the picoNewton range and we infer an effective charge of 12 e- per tubulin dimer under physiological conditions. PMID- 16690867 TI - Oceanographic basis of the global surface distribution of Prochlorococcus ecotypes. AB - By using data collected during a continuous circumnavigation of the Southern Hemisphere, we observed clear patterns in the population-genetic structure of Prochlorococcus, the most abundant photosynthetic organism on Earth, between and within the three Southern Subtropical Gyres. The same mechanisms that were previously invoked to account for the vertical distribution of ecotypes at local scales accounted for the global (horizontal) patterns we observed. Basin-scale and seasonal variations in the structure and strength of vertical stratification provide a basis for understanding large-scale horizontal distribution in genetic and physiological traits of Prochlorococcus, and perhaps of marine microbial communities in general. PMID- 16690868 TI - Ischemia opens neuronal gap junction hemichannels. AB - Neuronal excitotoxicity during stroke is caused by activation of unidentified large-conductance channels, leading to swelling and calcium dysregulation. We show that ischemic-like conditions [O(2)/glucose deprivation (OGD)] open hemichannels, or half gap junctions, in neurons. Hemichannel opening was indicated by a large linear current and flux across the membrane of small fluorescent molecules. Single-channel openings of hemichannels (530 picosiemens) were observed in OGD. Both the current and dye flux were blocked by inhibitors of hemichannels. Therefore, hemichannel opening contributes to the profound ionic dysregulation during stroke and may be a ubiquitous component of ischemic neuronal death. PMID- 16690869 TI - Hypothalamic mTOR signaling regulates food intake. AB - The mammalian Target of Rapamycin (mTOR) protein is a serine-threonine kinase that regulates cell-cycle progression and growth by sensing changes in energy status. We demonstrated that mTOR signaling plays a role in the brain mechanisms that respond to nutrient availability, regulating energy balance. In the rat, mTOR signaling is controlled by energy status in specific regions of the hypothalamus and colocalizes with neuropeptide Y and proopiomelanocortin neurons in the arcuate nucleus. Central administration of leucine increases hypothalamic mTOR signaling and decreases food intake and body weight. The hormone leptin increases hypothalamic mTOR activity, and the inhibition of mTOR signaling blunts leptin's anorectic effect. Thus, mTOR is a cellular fuel sensor whose hypothalamic activity is directly tied to the regulation of energy intake. PMID- 16690870 TI - Individual comparisons of the levels of (E)-3-methyl-2-hexenoic acid, an axillary odor-related compound, in Japanese. AB - The (E)-3-methyl-2-hexenoic acid (E3M2H), an axillary odor-related compound, is known to occur in Caucasians. The aims of this study were to clarify whether E3M2H contributes to axillary odor in Asians and to quantify and compare individual levels of E3M2H. Quantitative determination of E3M2H was performed by means of gas chromatography-mass spectrometry of sweat extracted from the axillary areas of T-shirts worn for 24 h by Japanese subjects. The amount of E3M2H was 15.9-34.6 nmol/ml in six of 30 subjects. Our method succeeded in quantitative analysis of E3M2H from axillary sweat collected individually; we also showed that E3M2H could be detected in Asians. This is the first report in which the amount of E3M2H in axillary sweat was quantified on an individual basis and compared to reveal individual differences. The results of this study indicate that E3M2H might contribute to axillary malodor in Asians as well as Caucasians. PMID- 16690871 TI - The capacity of humans to identify components in complex odor-taste mixtures. AB - Despite the fact that humans experience mixtures of odors and tastes each time they eat, little is known of their capacity to detect the individual components of foods. To investigate this capacity, 43 subjects were trained to identify three odors and three tastes and were required to indicate which of these could be identified in stimuli consisting of one to six components. Although the odor and taste components of most binary mixtures were identified, subjects encountered substantial difficulties with more complex mixtures with only two components being identified in the four- to six-component mixtures. In general, tastes were more easily identified than smells and were the only stimuli identified in the five- to six-component mixtures. Several mechanisms are proposed to account for the poor identification of components. PMID- 16690872 TI - Oxidized low-density lipoprotein correlates positively with toll-like receptor 2 and interferon regulatory factor-1 and inversely with superoxide dismutase-1 expression: studies in hypercholesterolemic swine and THP-1 cells. AB - BACKGROUND: Oxidized low-density lipoprotein (LDL) is associated with cardiovascular disease. Macrophages contribute to LDL oxidation, and oxidized LDL (oxLDL) affects macrophage function. We searched for the strongest gene correlates of oxLDL in macrophages in coronary plaques and studied the effect of oxLDL on their expression in THP-1 cells. METHODS AND RESULTS: Gene expression in macrophages isolated from coronary plaque macrophages from hypercholesterolemic swine was measured on Agilent Human cDNA microarrays. Compared with a universal reference, 1653 transcripts were deregulated. The expression of 11 genes correlated positively and the expression of 5 genes correlated negatively with plaque oxLDL. Interferon regulatory factor-1 (IRF1; R2 = 0.69) and toll-like receptor 2 (TLR2; R2 = 0.18) were the strongest positive correlates of oxLDL. Superoxide dismutase 1 (SOD1) was the strongest inverse correlate of oxLDL (R2 = 0.57). Immunohistochemical analysis showed colocalization of IRF1, TLR2, and SOD1 protein in macrophages and confirmed the RNA expression data. OxLDL-induced foam cell formation in THP-1 macrophages was associated with increased expression of IRF1 and TLR2 and decreased expression of SOD1. CONCLUSIONS: Our data support the hypothesis that oxLDL is a proinflammatory stimulus that induces the expression of TLR2 and IRF1, 2 important gene regulators of innate immune response, and inhibits the expression of the antioxidant SOD1. PMID- 16690873 TI - Elevated plasma active matrix metalloproteinase-9 level is associated with coronary artery in-stent restenosis. AB - OBJECTIVE: This study aimed to determine whether the plasma levels of matrix metalloproteinase-9 (MMP-9) or tissue inhibitor of metalloproteinases-1 (TIMP-1) were altered in patients with a history of symptomatic in-stent restenosis (ISR). METHODS AND RESULTS: A group of 158 patients with a history of ISR were compared with 128 symptom-free patients. Plasma samples and a detailed risk factor history were collected. Plasma samples were analyzed for pro-MMP-9 and latent MMP-9 and active MMP-9, latent MMP-3, and TIMP-1. Several variables were associated with ISR, including index coronary disease extent and severity (number of diseased vessels and American College of Cardiology/American Heart Association lesion classification), number, diameter, and total length of stent(s) inserted, and plasma high-density lipoprotein cholesterol. Plasma active MMP-9 (odds ratio, 1.96; 95% CI, 1.43 to 2.69) showed independent risk association with ISR. Patients with multiple sites of ISR had significantly higher levels of active MMP 9 compared with patients with only a single ISR lesion or no ISR. CONCLUSIONS: Plasma active MMP-9 levels may be a useful independent predictor of bare metal stent ISR. PMID- 16690874 TI - Gene variants of VAMP8 and HNRPUL1 are associated with early-onset myocardial infarction. AB - OBJECTIVE: Identify gene variants associated with early-onset myocardial infarction (MI). METHODS AND RESULTS: We tested 11 647 single-nucleotide polymorphisms (SNPs) for association with early-onset MI in a case-control study (study 1 200 cases, 262 controls). To reduce the number of false positives among the 666 SNPs that were nominally associated with early-onset MI (P<0.05) in study 1, we tested these SNPs in study 2 (434 cases, 504 controls). We found that 8 of the 666 SNPs were associated with early-onset MI in study 2 (P<0.05) and had the same risk alleles as in study 1. These 8 SNPs were then tested for association with early-onset MI in study 3 (187 cases, 434 controls). We found that a VAMP8 variant (P = 0.025; odds ratio [OR], 1.75; CI, 1.17 to 2.62) and an HNRPUL1 variant (P = 0.0043; OR, 1.92; CI, 1.28 to 2.86) were associated with early-onset MI (nominal P<0.05; false discovery rate <10%) and had the same risk alleles in all 3 studies. CONCLUSIONS: Variants in 2 genes were associated with early-onset MI: VAMP8, which is involved in platelet degranulation, and HNRPUL1, which encodes a ribonuclear protein. The identification of these variants could improve understanding of disease mechanisms and suggest novel drug targets. PMID- 16690875 TI - Differentiation of lymphatic endothelial cells from embryonic stem cells on OP9 stromal cells. AB - OBJECTIVE: The discovery of vascular endothelial growth factor C (VEGF-C) and VEGF receptor-3 (VEGFR-3) has started to provide an understanding of the molecular mechanisms of lymphangiogenesis. The homeobox gene prox1 has been proven to specify lymphatic endothelial cells (ECs) from blood ECs. We investigated the process of lymphatic EC (LEC) differentiation using embryonic stem (ES) cells. METHODS AND RESULTS: VEGFR-2+ cells derived from ES cells differentiated into LECs at day 3 on OP9 stromal cells defined by the expression of prox1, VEGFR-3, and another lymphatic marker podoplanin. VEGFR-2+ cells gave rise to LYVE-1+ embryonic ECs, which were negative for prox1 on day 1 but turned to prox1+ LECs by day 3. VEGFR-3-Fc or Tie2-Fc, sequestering VEGF-C or angiopoietin1 (Ang1), suppressed colony formation of LECs on OP9 cells. However, addition of VEGF-C and Ang1 in combination with VEGF to the culture of VEGFR-2+ cells on collagen-coated dishes failed to induce LECs. LEC-inducing activity of OP9 cells was fully reproduced on paraformaldehyde-fixed OP9 cells with the conditioned medium. CONCLUSIONS: We succeeded in differentiating LECs from ES cells and revealed the requirements of VEGF-C, Ang1, and other unknown factors for LEC differentiation. PMID- 16690876 TI - Matrix metalloproteinase inhibition attenuates aortic calcification. AB - OBJECTIVE: Arterial calcification has been associated with matrix metalloproteinase (MMP)-mediated elastin degradation. In this study, we investigated whether inhibiting MMP activity could reduce calcium accumulation in rodent models of aortic calcification. METHODS AND RESULTS: Aortic calcification was first induced in male Sprague-Dawley rats by administration of vitamin D3. Treatment with doxycycline decreased aortic calcium and phosphorus accumulation, and it reduced aortic gelatinase levels; however, it also prevented the bone resorption associated with high doses of vitamin D3. Using an in vivo model of localized aortic calcification, systemic doxycycline treatment reduced aortic calcium accumulation without affecting serum calcium levels, suggesting a more specific effect of doxycycline in the arterial wall. In organ culture, doxycycline limited aortic calcification caused by exposure to alkaline phosphatase and inorganic phosphate. When GM6001, a synthetic and specific inhibitor of MMPs, was used instead of doxycycline, it had a similar effect. In vivo, periadventitial delivery of GM6001 to calcifying arteries significantly reduced calcification compared with controls. CONCLUSIONS: These results suggest that MMPs are involved in aortic calcification, and inhibiting MMP activity could reduce calcium accumulation in the arterial wall. PMID- 16690877 TI - Matrix metalloproteinase 2 activation of transforming growth factor-beta1 (TGF beta1) and TGF-beta1-type II receptor signaling within the aged arterial wall. AB - OBJECTIVE: To study matrix metalloproteinase 2 (MMP-2) effects on transforming growth factor-beta1 (TGF-beta1) activation status and downstream signaling during arterial aging. METHODS AND RESULTS: Western blotting and immunostaining showed that latent and activated TGF-beta1 are markedly increased within the aorta of aged Fisher 344 cross-bred Brown Norway (30 months of age) rats compared with adult (8 months of age) rats. Aortic TGF-beta1-type II receptor (TbetaRII), its downstream molecules p-similar to mad-mother against decapentaplegic (SMAD)2/3 and SMAD4, fibronectin, and collagen also increased with age. Moreover, TGF-beta1 staining is colocalized with that of activated MMP-2 within the aged arterial wall and vascular smooth muscle cell (VSMC) in vitro, and this physical association was confirmed by coimmunoprecipitation. Incubation of young aortic rings ex vivo or VSMCs in vitro with activated MMP-2 enhanced active TGF-beta1, collagen, and fibronectin expression to the level of untreated old counterparts, and this effect was abolished via inhibitors of MMP-2. Interestingly, in old untreated rings or VSMCs, the increased TGF-beta1, fibronectin, and collagen were also substantially reduced by inhibition of MMP-2. CONCLUSIONS: Active TGF-beta1, its receptor, and receptor-mediated signaling increase within the aortic wall with aging. TGF-beta1 activation is dependent, in part at least, by a concomitant age-associated increase in MMP-2 activity. Thus, MMP-2-activated TGF-beta1, and subsequently TbetaRII signaling, is a novel molecular mechanism for arterial aging. PMID- 16690878 TI - Effects of congestive heart failure on Ca2+ handling in skeletal muscle during fatigue. AB - Skeletal muscle weakness and decreased exercise capacity are major symptoms reported by patients with congestive heart failure (CHF). Intriguingly, these skeletal muscle symptoms do not correlate with the decreased heart function. This suggests that CHF leads to maladaptive changes in skeletal muscles, and as reported most markedly in slow-twitch muscles. We used rats at 6 weeks after infarction to measure expression of key proteins involved in SR Ca(2+) release and uptake in slow-twitch soleus muscles. We also measured force and myoplasmic free [Ca(2+)] ([Ca(2+)](i)) in intact single fibers of soleus muscles. CHF rats showed clear signs of severe cardiac dysfunction with marked increases in heart weight and left ventricular end-diastolic pressure compared with sham operated rats (Sham). There were small, but significant, changes in the content of proteins involved in cellular Ca(2+) handling in CHF muscles: slight increases in SR Ca(2+) release channels (ie, the ryanodine receptors) and in SR Ca(2+)-ATPase. Tetanic force and [Ca(2+)](i) were not significantly different between CHF and Sham soleus fibers under resting conditions. However, during the stimulation period there was a decrease in tetanic force without changes in [Ca(2+)](i) in CHF fibers that was not observed in Sham fibers. The fatigue-induced changes recovered rapidly. We conclude that CHF soleus fibers fatigue more rapidly than Sham fibers because of a reversible fatigue-induced decrease in myofibrillar function. PMID- 16690879 TI - Induction of cardiogenesis in embryonic stem cells via downregulation of Notch1 signaling. AB - Embryonic stem cells represent an attractive source of cardiomyocytes for cell replacement therapies. However, before embryonic stem cells can be successfully used for the treatment of cardiac diseases, the precise molecular mechanisms that underlie their cardiogenic differentiation must be identified. A network of intrinsic and extrinsic factors regulates embryonic stem cell self-renewal and differentiation into a variety of different cell lineages. Here, we show that Notch signaling takes place in some but not all embryonic stem cells and that the Notch pathway is shut down during the course of differentiation concomitantly with downregulation of Notch receptor and ligand expression. Moreover, gain- and loss-of-function experiments for Notch signaling components show that this pathway is a crucial regulator of cardiomyocyte differentiation within ES cells. Differentiation of ES cells into cardiomyocytes is favored by inactivation of the Notch1 receptor, whereas endogenous Notch signaling promotes differentiation of ES cells into the neuronal lineage. We conclude that Notch signaling influences the cell fate decision between mesodermal and the neuroectodermal cell fates during embryonic stem cell differentiation. These findings should help to optimize the production of specific cell types via modulation of the Notch pathways and, in particular, to improve the production of embryonic stem cell derived cardiomyocytes. PMID- 16690880 TI - Heteromultimeric TRPC6-TRPC7 channels contribute to arginine vasopressin-induced cation current of A7r5 vascular smooth muscle cells. AB - The molecular identity of receptor-operated, nonselective cation channels (ROCs) of vascular smooth muscle (VSM) cells is not known for certain. Mammalian homologues of the Drosophila canonical transient receptor potential channels (TRPCs) are possible candidates. This study tested the hypothesis that heteromultimeric TRPC channels contribute to ROC current of A7r5 VSM cells activated by [Arg(8)]-vasopressin. A7r5 cells expressed transcripts encoding TRPC1, TRPC4beta, TRPC6, and TRPC7. TRPC4, TRPC6, and TRPC7 protein expression was confirmed by immunoblotting and association of TRPC6 with TRPC7, but not TRPC4beta, was detected by coimmunoprecipitation. The amplitude of arginine vasopressin (AVP)-induced ROC current was suppressed by dominant-negative mutant TRPC6 (TRPC6(DN)) but not TRPC5 (TRPC5(DN)) mutant subunit expression. These data indicate a role for TRPC6- and/or TRPC7-containing channels and rule a more complex subunit composition including TRPC1 and TRPC4. Increasing extracellular Ca(2+) concentration ([Ca(2+)](o)) from 0.05 to 1 mmol/L suppressed currents owing to native, TRPC7, and heteromultimeric TRPC6-TRPC7 channels, but not TRPC6 current, which was slightly enhanced. The relative changes in native and heteromultimeric TRPC6-TRPC7 current amplitudes for [Ca(2+)](o) between approximately 0.01 and 1 mmol/L were identical, but the changes in homomultimeric TRPC6 and TRPC7 currents were significantly less and greater, respectively, compared with the native channels. Taken together, the data provide biochemical and functional evidence supporting the view that heteromultimeric TRPC6-TRPC7 channels contribute to receptor-activated, nonselective cation channels of A7r5 VSM cells. PMID- 16690881 TI - Common protective and diverse smooth muscle cell effects of AAV-mediated angiopoietin-1 and -2 expression in rat cardiac allograft vasculopathy. AB - Angiopoietin-1 (Ang1) and Ang2 regulate the maintenance of normal vasculature by direct endothelial and indirect smooth muscle cell (SMC) effects. Dysfunction of vascular wall cells is considered central in cardiac allograft vasculopathy (CAV), where inflammation and arterial injury initiate subsequent intimal SMC proliferation. In this study, we investigated the effect of exogenous Ang1 and Ang2 in chronically rejecting rat cardiac allografts by intracoronary adeno associated virus (AAV)-mediated gene transfer. Bioluminescent imaging of AAV transfected syngeneic grafts revealed gradual and stable transgene expression in graft cardiomyocytes. In cardiac allografts, both AAV-Ang1 and AAV-Ang2 decreased inflammation and increased antiapoptotic Bcl-2 mRNA and Bcl-2/Bax ratio at 8 weeks. Only AAV-Ang2 decreased the development of CAV, whereas AAV-Ang1 activated arterial SMC and increased PDGF-A mRNA in the allograft. Collectively, our results show that exogenous Ang1 and Ang2 have similar antiinflammatory and antiapoptotic effects in cardiac allografts. Prolonged AAV-mediated Ang1 transgene expression also induced SMC activation, whereas AAV-Ang2 lacked the SMC activating effects and decreased CAV. Our results thus highlight the common protective and diverse SMC effects of Ang1 and Ang2 in cardiac allograft microenvironment and the importance of timing of angiopoietins to achieve therapeutic effects. PMID- 16690882 TI - Bone marrow stem cells prevent left ventricular remodeling of ischemic heart through paracrine signaling. AB - In this study, we hypothesized that bone marrow stem cells (BMSCs) protect ischemic myocardium through paracrine effects that can be further augmented with preconditioning. In in vitro experiments, cell survival factors such as Akt and eNOS were significantly increased in BMSCs following anoxia. In the second series of experiments following coronary ligation in mice, left ventricles were randomly injected with the following: DMEM (G-1), BMSCs (G-2), and preconditioned BMSCs (G 3). Four days after myocardial infarction, BMSCs were observed within injured myocardium in G-2 and G-3. Apoptotic cardiomyocytes within periinfarct area were significantly reduced in G-3. Four weeks after myocardial infarction, smaller left ventricular (LV) dimension and increased LV ejection fraction were observed in G-3. Infarct area was significantly reduced in G-3. However, GFP+ cardiomyocytes were observed in low numbers within periinfarct area in G-2 and G 3. In conclusion, BMSCs secreted cell survival factors under ischemia, and they prevented apoptosis in cardiomyocytes adjacent to the infarcted area. Preconditioning of BMSCs enhanced their survival and ability to attenuate LV remodeling, which was attributable, in part, to paracrine effects. PMID- 16690883 TI - Identification of a novel peptide that interferes with the chemical regulation of connexin43. AB - The carboxyl-terminal domain of connexin43 (Cx43CT) is involved in various intra- and intermolecular interactions that regulate gap junctions. Here, we used phage display to identify novel peptidic sequences that bind Cx43CT and modify Cx43 regulation. We found that Cx43CT binds preferentially to peptides containing a sequence RXP, where X represents any amino acid and R and P correspond to the amino acids arginine and proline, respectively. A biased "RXP library" led to the identification of a peptide (dubbed "RXP-E") that bound Cx43CT with high affinity. Nuclear magnetic resonance data showed RXP-E-induced shifts in the resonance peaks of residues 343 to 346 and 376 to 379 of Cx43CT. Patch-clamp studies revealed that RXP-E partially prevented octanol-induced and acidification induced uncoupling in Cx43-expressing cells. Moreover, RXP-E increased mean open time of Cx43 channels. The full effect of RXP-E was dependent on the integrity of the CT domain. These data suggest that RXP-based peptides could serve as tools to help determine the role of Cx43 as a regulator of function in conditions such as ischemia-induced arrhythmias. PMID- 16690884 TI - Bradycardia and slowing of the atrioventricular conduction in mice lacking CaV3.1/alpha1G T-type calcium channels. AB - The generation of the mammalian heartbeat is a complex and vital function requiring multiple and coordinated ionic channel activities. The functional role of low-voltage activated (LVA) T-type calcium channels in the pacemaker activity of the sinoatrial node (SAN) is, to date, unresolved. Here we show that disruption of the gene coding for CaV3.1/alpha1G T-type calcium channels (cacna1g) abolishes T-type calcium current (I(Ca,T)) in isolated cells from the SAN and the atrioventricular node without affecting the L-type Ca2+ current (I(Ca,L)). By using telemetric electrocardiograms on unrestrained mice and intracardiac recordings, we find that cacna1g inactivation causes bradycardia and delays atrioventricular conduction without affecting the excitability of the right atrium. Consistently, no I(Ca,T) was detected in right atrium myocytes in both wild-type and CaV3.1(-/-) mice. Furthermore, inactivation of cacna1g significantly slowed the intrinsic in vivo heart rate, prolonged the SAN recovery time, and slowed pacemaker activity of individual SAN cells through a reduction of the slope of the diastolic depolarization. Our results demonstrate that CaV3.1/T-type Ca2+ channels contribute to SAN pacemaker activity and atrioventricular conduction. PMID- 16690885 TI - Angiopoietin and vascular remodeling: the data at face value. PMID- 16690886 TI - Response to arachidonate 5-lipoxygenase variants in atherosclerosis, obesity, and bone traits. PMID- 16690887 TI - MCP-1 induces a novel transcription factor with proapoptotic activity. PMID- 16690888 TI - When is a fly in the ointment a solution and not a problem? PMID- 16690889 TI - Closing the cycle: Skp2 modulates cyclic nucleotides antiproliferative effects. PMID- 16690890 TI - Therapeutic angiogenesis: another passing phase? PMID- 16690891 TI - A farewell kiss triggers a broken heart? PMID- 16690892 TI - Viewing a stressful episode of ER: is ATF6 the triage nurse? PMID- 16690893 TI - ACE polymorphisms. AB - Angiotensin converting enzyme (ACE) plays an essential role in two physiological systems, one leading to the production of angiotensin II and the other to the degradation of bradykinin. The wide distribution and multifunctional properties of these peptides suggest that ACE could be involved in various pathophysiological conditions. The discovery that ACE levels are under genetic control ushered in a new era of investigation; most studies focused on an insertion/deletion (I/D) polymorphism in intron 16 of the ACE gene as a marker for a functional polymorphism. Recently, many single nucleotide polymorphisms were detected in the gene and the search for the locations of functional polymorphisms became a topic of extensive investigation. Nevertheless, association studies on the I/D polymorphism and clinical outcomes continued, mostly with conflicting results. This article reviews the current state of knowledge regarding ACE polymorphisms and suggests that a functional polymorphism is most likely located between intron 18 and the 3' UTR. The potential existence of another functional polymorphism in the 5' UTR, however, cannot be excluded. This review also presents an overview of ACE function in different pathophysiological systems, and summarizes previous reports on ACE and clinical outcomes. Although findings on the I/D polymorphism and disorders like diabetic nephropathy and Alzheimer disease can be considered conclusive, reports on most of the cardiovascular phenotypes are still controversial. Genotypic and phenotypic misclassifications, insufficient power in some studies, and the presence of interaction with other genes or environmental factors are possible explanations for the contradictory findings. PMID- 16690894 TI - For rich and poor, the message is still "dial 9-1-1": but is it getting through? PMID- 16690896 TI - Increased brain expression of matrix metalloproteinase-9 after ischemic and hemorrhagic human stroke. AB - BACKGROUND AND PURPOSE: Abnormal expression of some matrix metalloproteinases (MMP) has shown to play a deleterious role in brain injury in experimental models of cerebral ischemia. We aimed to investigate MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in brain parenchyma in both ischemic and hemorrhagic strokes. METHODS: Postmortem fresh brain tissue from 6 ischemic and 8 hemorrhagic stroke patients was obtained within the first 6 hours after death. Finally, 78 brain tissue samples from different areas (infarct, peri-infarct, perihematoma and contralateral hemisphere) were studied. To quantify gelatinase content we performed gelatin zymograms that were confirmed by Western Blot Analysis, immunohistochemistry to localize MMP source, and in situ zymography to detect gelatinase activity. RESULTS: Among ischemic cases, gelatin zymography showed increased MMP-9 content in infarct core although peri-infarct tissue presented also higher levels than contralateral hemisphere (P<0.0001 and P=0.042, respectively). Within infarct core, MMP-9 was mainly located around blood vessels, associated to neutrophil infiltration and activated microglial cells. In peri-infarct areas the major source of MMP-9 were microglial cells. Tissue around intracranial hemorrhage also displayed higher MMP-9 levels than contralateral hemisphere (P=0.008) in close relationship with glial cells. MMP-2 was constitutively expressed and remained invariable in different brain areas. CONCLUSIONS: Our results demonstrate in situ higher levels of MMP-9 in human brain tissue after ischemic and hemorrhagic stroke, suggesting a contribution of MMP-9 to ischemic brain injury and perihematoma edema. PMID- 16690897 TI - A discriminative prediction model of neurological outcome for patients undergoing surgery of brain arteriovenous malformations. AB - BACKGROUND AND PURPOSE: To develop and validate a discriminative model for predicting neurological morbidity after brain arteriovenous malformation (bAVM) surgery. METHODS: Of 233 consecutive, prospectively enrolled patients undergoing bAVM surgery, the first 175 were used to derive, and the last 58 to validate, the prediction model. Demographic and angiographic factors were related to modified Rankin Scale scores assigned before, within 72 hours, at 7 days and at > or =1 year after surgery to seek predictors of postoperative neurological deficits (modified Rankin Scale score > or =3). These factors included nidus size, eloquence, venous drainage, diffuseness, white matter configuration, arterial perforator supply and associated aneurysms. RESULTS: Brain eloquence, diffuse nidus and deep venous drainage were significant predictors of early disabling neurological deficits (odds ratios of 4.33, 3.49 and 2.38, respectively). The rounded odds ratios form a weighted 9-point prediction model (maximum scores for eloquence+diffuseness+deep drainage=4+3+2). The score discriminated the probability of experiencing both early (first week) and permanently (at > or =1 year) disabling neurological deficits as follows: 0 to 2: 1.8%, 3 to 5: 17.4%, 6 to 7: 31.6%, >7: 52.9% for early and 0 to 2: 1.8%, 3 to 5: 4.4%, 6 to 7: 18.4%, >7: 32.4% for permanently disabling outcomes. The discrimination of the model was 0.80 with 2.8% optimism. Validation in the second patient cohort revealed good performance at risk stratification. CONCLUSIONS: Relative weights assigned to brain eloquence, diffuse nidus morphology and deep venous drainage of a bAVM provide a simple and discriminative prediction model for neurological outcome after bAVM surgery. PMID- 16690898 TI - Community socioeconomic status and prehospital times in acute stroke and transient ischemic attack: do poorer patients have longer delays from 911 call to the emergency department? AB - BACKGROUND AND PURPOSE: Timely access to medical treatment is critical for patients with acute stroke because acute therapies must be given very quickly after symptom onset. We examined the effect of socioeconomic status on prehospital delays in stroke and transient ischemic attack (TIA) patients within a large, biracial population. METHODS: By screening all local hospital ICD-9 codes 430 to 436, all stroke and TIA patients were identified during the calendar year of 1999. Cases must have used emergency medical services (EMS), lived at home, had their stroke at home, and had documented times of the 911 call and arrival to the emergency department. Socioeconomic status was estimated using economic data regarding the geocoded home residence census tract. RESULTS: Only 38% of stroke and TIA patients used EMS. There were 978 cases of stroke and TIA included in this analysis. The mean times were call to arrival on scene 6.5 minutes, on-scene time 14.1 minutes, and transport time 13.1 minutes. Lower community socioeconomic status was associated with all 3 EMS time intervals; however, all time differences were small: the largest difference was 5 minutes. CONCLUSIONS: Within our population, living in a poorer area does not appear to delay access to acute care for stroke in a clinically significant way. We did find small, statistically significant delays in prehospital times that were associated with poorer communities, black race, and increasing age. However, delays related to public recognition of stroke symptoms, and limited use of 911, are likely much more important than these small delays that occur with EMS systems. PMID- 16690899 TI - Epidemiology of aphasia attributable to first ischemic stroke: incidence, severity, fluency, etiology, and thrombolysis. AB - BACKGROUND AND PURPOSE: In a geographically defined population, we assessed incidence and determinants of aphasia attributable to first-ever ischemic stroke (FEIS). METHODS: A 1-year prospective, population-based study among the permanent residents of the canton Basle City, Switzerland, was performed using multiple overlapping sources of information. RESULTS: Among 188,015 inhabitants, 269 patients had FEIS, of whom 80 (30%; 95% CI, 24 to 36) had aphasia. The overall incidence rate of aphasia attributable to FEIS amounted to 43 per 100,000 inhabitants (95% CI, 33 to 52). Aphasic stroke patients were older than nonaphasic patients. The risk of aphasia attributable to FEIS increased by 4% (95% CI, 1% to 7%), and after controlling for atrial fibrillation, by 3% (95% CI, 1% to 7%) with each year of patients' age. Gender had no effect on incidence, severity, or fluency of aphasia. Cardioembolism was more frequent in aphasic stroke patients than in nonaphasic ones (odds ratio [OR], 1.85; 95% CI, 1.07 to 3.20). Aphasic patients sought medical help earlier than nonaphasic stroke patients. Still, after controlling for stroke onset-assessment interval, aphasic stroke patients were more likely to receive thrombolysis than nonaphasics (OR, 3.5; 95% CI, 1.12 to 10.96). CONCLUSIONS: Annually, 43 of 100,000 inhabitants had aphasia resulting from first ischemic stroke. Advancing age and cardioembolism were associated with an increased risk for aphasia. Severity and fluency of aphasia were not affected by demographic variables. PMID- 16690900 TI - Proportions of stroke subtypes among men and women > or =40 years of age in an urban Japanese city in 1992, 1997, and 2002. AB - BACKGROUND AND PURPOSE: Higher proportions of hemorrhagic stroke and lacunar infarction were reported in rural Japan compared with those in Western countries. We examined the relative proportions of stroke subtypes in an urban Japanese city where westernized lifestyles are more common than in rural areas. METHODS: Stroke registration was performed in 1992, 1997, and 2002 for residents > or =40 years of age who were admitted with acute strokes to all of the 10 hospitals with > or =90 beds in Yao City, Osaka, Japan. Strokes were classified as intraparenchymal hemorrhage, subarachnoid hemorrhage, or ischemic strokes (embolic infarction, large-artery occlusive infarction, lacunar infarction, and unclassified thrombotic infarction) by criteria using computed tomography or MRI. RESULTS: A total of 650 first-ever strokes were registered. The age-adjusted proportion of each stroke subtype was not significantly different among the 3 study periods in both men and women. Throughout the 3 periods, intraparenchymal hemorrhage, subarachnoid hemorrhage, and ischemic stroke accounted for 26%, 7%, and 65% in men, respectively. In women, the respective proportions were 29%, 21%, and 44%. The proportion of each subtype for total ischemic strokes was as follows: 51% to 61% lacunar infarction, 25% to 26% large-artery occlusive infarction, and 11% to 17% embolic infarction. CONCLUSIONS: Our study showed that hemorrhagic stroke represented a large proportion of all strokes, especially among women, and lacunar infarction was the most common subtype of ischemic stroke among both men and women in Yao City, which differed from findings in Western countries. PMID- 16690901 TI - D-dimer as an independent predictor for poor outcome after aneurysmal subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: After aneurysmal subarachnoid hemorrhage (SAH), elevated D-dimer levels have been associated with poor clinical condition and outcome. We tested prospectively whether D-dimer values affect outcome after SAH independently of severity of bleeding. METHODS: Previous diseases, and clinical as well as radiological variables, were recorded for 136 patients with SAH admitted within 48 hours after bleeding. Plasma D-dimer was measured in the morning after aneurysm occlusion and at discharge 10 to 12 days after SAH. Factors predicting poor outcome according to the Glasgow Outcome Scale at 3 months after SAH and appearance of cerebral infarction were tested with multiple logistic regression. RESULTS: Patients with poor outcome had higher D-dimer values than did those with favorable outcome: after surgery, a median 1250 (25th and 75th percentiles 675 and 2900) microg/L versus 720 (350 and 1119) microg/L (P=0.001); and at discharge, 1150 (624 and 2875) microg/L versus 360 (330 and 600) microg/L (P<0.001), respectively. In repeated-measures ANOVA, D-dimer decreased more rapidly (P=0.022) in those with favorable outcome. After simultaneous adjustment for several factors affecting outcome, plasma D-dimer after surgery remained a significant predictor for poor outcome (odds ratio, 1.63 per mg/L; 95% CI, 1.03 to 2.60; P=0.038) but neither for delayed ischemia nor, on follow-up computed tomography in survivors, for cerebral infarction. CONCLUSIONS: Elevated plasma D-dimer after admission independently predicts poor outcome, suggesting that prolonged excess thrombin generation may impair outcome. Repeated high plasma D-dimer values can be useful in discovering patients at increased risk for poor outcome. PMID- 16690902 TI - Socioeconomic status and stroke incidence in the US elderly: the role of risk factors in the EPESE study. AB - BACKGROUND AND PURPOSE: This study assesses the effect of socioeconomic status on stroke incidence in the elderly, and the contribution of risk factors to stroke disparities. METHODS: Data comprised a sample of 2812 men and women aged 65 years and over from the New Haven cohort of the Established Populations for the Epidemiologic Studies of the Elderly. Individuals provided baseline information on demographics, functioning, cardiovascular and psychosocial risk factors in 1982 and were followed for 12 years. Proportional hazard models were used to model survival from initial interview to first fatal or nonfatal stroke. RESULTS: Two hundred and seventy subjects developed incident stroke. At ages 65 to 74, lower socioeconomic status was associated with higher stroke incidence for both education (HR(lowest/highest)=2.07, 95% CI, 1.04 to 4.13) and income (HR(lowest/highest)=2.08, 95% CI, 1.01 to 4.27). Adjustment for race, diabetes, depression, social networks and functioning attenuated hazard ratios to a nonsignificant level, whereas other risk factors did not change associations significantly. Beyond age 75, however, stroke rates were higher among those with the highest education (HR(lowest/highest)=0.42, 95% CI, 0.22 to 0.79) and income (HR(lowest/highest)=0.43, 95% CI, 0.22 to 0.86), which remained largely unchanged after adjustment for risk factors. CONCLUSIONS: We observed substantial socioeconomic disparities in stroke at ages 65 to 74, whereas a crossover of the association occurred beyond age 75. Policies to improve social and economic resources at early old age, and interventions to improve diabetes management, depression, social networks and functioning in the disadvantaged elderly can contribute to reduce stroke disparities. PMID- 16690903 TI - NHE2 is the main apical NHE in mouse colonic crypts but an alternative Na+ dependent acid extrusion mechanism is upregulated in NHE2-null mice. AB - The mechanism of apical Na(+)-dependent H(+) extrusion in colonic crypts is controversial. With the use of confocal microscopy of the living mouse distal colon loaded with BCECF or SNARF-5F (fluorescent pH sensors), measurements of intracellular pH (pH(i)) in epithelial cells at either the crypt base or colonic surface were reported. After cellular acidification, the addition of luminal Na(+) stimulated similar rates of pH(i) recovery in cells at the base of distal colonic crypts of wild-type or Na(+)/H(+) exchanger isoform 2 (NHE2)-null mice. In wild-type crypts, 20 microM HOE694 (NHE2 inhibitor) blocked 68-75% of the pH(i) recovery rate, whereas NHE2-null crypts were insensitive to HOE694, the NHE3-specific inhibitor S-1611 (20 microM), or the bicarbonate transport inhibitor 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS; 1 mM). A general NHE inhibitor, 5-(N-ethyl-N-isopropyl)amiloride (EIPA; 20 microM), inhibited pH(i) recovery in NHE2-null mice (46%) but less strongly than in wild type mice (74%), suggesting both EIPA-sensitive and -insensitive compensatory mechanisms. Transepithelial Na(+) leakage followed by activation of basolateral NHE1 could confound the outcomes; however, the rates of Na(+)-dependent pH(i) recovery were independent of transepithelial leakiness to lucifer yellow and were unchanged in NHE1-null mice. NHE2 was immunolocalized on apical membranes of wild type crypts but not NHE2-null tissue. NHE3 immunoreactivity was near the colonic surface but not at the crypt base in NHE2-null mice. Colonic surface cells from wild-type mice demonstrated S1611- and HOE694-sensitive pH(i) recovery in response to luminal sodium, confirming a functional role for both NHE3 and NHE2 at this site. We conclude that constitutive absence of NHE2 results in a compensatory increase in a Na(+)-dependent, EIPA-sensitive acid extruder distinct from NHE1, NHE3, or SITS-sensitive transporters. PMID- 16690904 TI - Highly efficient gluten degradation with a newly identified prolyl endoprotease: implications for celiac disease. AB - Celiac disease is a T cell-driven intolerance to wheat gluten. The gluten-derived T cell epitopes are proline-rich and thereby highly resistant to proteolytic degradation within the gastrointestinal tract. Oral supplementation with prolyl oligopeptidases has therefore been proposed as a potential therapeutic approach. The enzymes studied, however, have limitations as they are irreversibly inactivated by pepsin and acidic pH, both present in the stomach. As a consequence, these enzymes will fail to degrade gluten before it reaches the small intestine, the site where gluten induces inflammatory T cell responses that lead to celiac disease. We have now determined the usefulness of a newly identified prolyl endoprotease from Aspergillus niger for this purpose. Gluten and its peptic/tryptic digest were treated with prolyl endoprotease, and the destruction of the T cell epitopes was tested using mass spectrometry, T cell proliferation assays, ELISA, reverse-phase HPLC, SDS-PAGE, and Western blotting. We observed that the A. niger prolyl endoprotease works optimally at 4-5 pH, remains stable at 2 pH, and is completely resistant to digestion with pepsin. Moreover, the A. niger-derived enzyme efficiently degraded all tested T cell stimulatory peptides as well as intact gluten molecules. On average, the endoprotease from A. niger degraded gluten peptides 60 times faster than a prolyl oligopeptidase. Together these results indicate that the enzyme from A. niger efficiently degrades gluten proteins. Future studies are required to determine if the prolyl endoprotease can be used as an oral supplement to reduce gluten intake in patients. PMID- 16690905 TI - Neurophysiological evaluation of healthy human anorectal sensation. AB - Patients with functional gastrointestinal disorders often demonstrate abnormal visceral sensation. Currently, rectal sensation is assessed by manual balloon distension or barostat. However, neither test is adaptable for use in the neurophysiological characterization of visceral afferent pathways by sensory evoked potentials. The aim of this study was to assess the reproducibility and quality of sensation evoked by electrical stimulation (ES) and rapid balloon distension (RBD) in the anorectum and to apply the optimum stimulus to examine the visceral afferent pathway with rectal evoked potentials. Healthy subjects (n = 8, median age 33 yr) were studied on three separate occasions. Variability, tolerance, and stimulus characteristics were assessed with each technique. Overall ES consistently invoked pain and was chosen for measuring rectal evoked potential whereas RBD in all cases induced the strong urge to defecate. Rectal intraclass correlation coefficient (ICC) for ES and RBD (0.82 and 0.72, respectively) demonstrated good reproducibility at pain/maximum tolerated volume but not at sensory threshold. Only sphincter ICC for ES at pain showed acceptable between-study reproducibility (ICC 0.79). Within studies ICC was good (>0.6) for anorectal ES and RBD at both levels of sensation. All subjects reported significantly more unpleasantness during RBD than ES (P < 0.01). This study demonstrates that ES and RBD are similarly reproducible. However, the sensations experienced with each technique differed markedly, probably reflecting differences in peripheral and/or central processing of the sensory input. This is of relevance in interpreting findings of neuroimaging studies of anorectal sensation and may provide insight into the physiological characteristics of visceral afferent pathways in health and disease. PMID- 16690906 TI - Nidovirus transcription: how to make sense...? AB - Many positive-stranded RNA viruses use subgenomic mRNAs to express part of their genetic information. To produce structural and accessory proteins, members of the order Nidovirales (corona-, toro-, arteri- and roniviruses) generate a 3' co terminal nested set of at least three and often seven to nine mRNAs. Coronavirus and arterivirus subgenomic transcripts are not only 3' co-terminal but also contain a common 5' leader sequence, which is derived from the genomic 5' end. Their synthesis involves a process of discontinuous RNA synthesis that resembles similarity-assisted RNA recombination. Most models proposed over the past 25 years assume co-transcriptional fusion of subgenomic RNA leader and body sequences, but there has been controversy over the question of whether this occurs during plus- or minus-strand synthesis. In the latter model, which has now gained considerable support, subgenomic mRNA synthesis takes place from a complementary set of subgenome-size minus-strand RNAs, produced by discontinuous minus-strand synthesis. Sense-antisense base-pairing interactions between short conserved sequences play a key regulatory role in this process. In view of the presumed common ancestry of nidoviruses, the recent finding that ronivirus and torovirus mRNAs do not contain a common 5' leader sequence is surprising. Apparently, major mechanistic differences must exist between nidoviruses, which raises questions about the functions of the common leader sequence and nidovirus transcriptase proteins and the evolution of nidovirus transcription. In this review, nidovirus transcription mechanisms are compared, the experimental systems used are critically assessed and, in particular, the impact of recently developed reverse genetic systems is discussed. PMID- 16690907 TI - Silencing T cells or T-cell silencing: concepts in virus-induced immunosuppression. AB - The ability to evade or suppress the host's immune response is a property of many viruses, indicating that this provides an advantage for the pathogen to spread efficiently or even to establish a persistent infection. The type and complexity of its genome and cell tropism but also its preferred type of host interaction are important parameters which define the strategy of a given virus to modulate the immune system in an optimal manner. Because they take a central position in any antiviral defence, the activation and function of T cells are the predominant target of many viral immunosuppressive regimens. In this review, two different strategies whereby this could be achieved are summarized. Retroviruses can infect professional antigen-presenting cells and impair their maturation and functional properties. This coincides with differentiation and expansion of silencing T cells referred to as regulatory T cells with suppressive activity, mainly to CD8+ effector T cells. The second concept, outlined for measles virus, is a direct, contact-mediated silencing of T cells which acquire a transient paralytic state. PMID- 16690908 TI - The immune response during hepatitis B virus infection. AB - Hepatitis B virus (HBV) is a major cause of chronic liver inflammation worldwide. Recent knowledge of the virological and immunological events secondary to HBV infection has increased our understanding of the mechanisms involved in viral clearance and persistence. In this review, how the early virological and immunological events might influence the development of a coordinate activation of adaptive immunity necessary to control HBV infection is analysed. The mechanism(s) by which high levels of viral antigens, liver immunological features, regulatory cells and dendritic cell defects might maintain the HBV specific immunological collapse, typical of chronic hepatitis B patients, is also examined. PMID- 16690909 TI - Vaccinia virus strain Western Reserve protein B14 is an intracellular virulence factor. AB - A characterization of the B14R gene from Vaccinia virus (VACV) strain Western Reserve (WR) is presented. Computational analyses of the B14R gene indicated high conservation in orthopoxviruses but no orthologues outside the Poxviridae. To characterize the B14 protein, the B14R gene was expressed in Escherichia coli and recombinant protein was purified and used to generate a rabbit polyclonal antiserum. This antiserum recognized a 15 kDa cytoplasmic protein in mammalian cells that were transfected with the B14R gene or infected with VACV WR, but not from cells infected with a VACV mutant (vdeltaB14) from which the B14R gene was deleted. Compared to wild-type and revertant virus controls, vdeltaB14 had normal growth kinetics in cell culture. The virulence of vdeltaB14 was assessed in two in vivo models. Mice infected intranasally with vdeltaB14 had similar weight loss compared to the controls, but in C57BL/6 mice infected intradermally vdeltaB14 induced a smaller lesion size compared with controls. Moreover, intradermal infection with vdeltaB14 caused an increased infiltration of cells into the infected lesion despite the smaller lesion size. Therefore, B14 is an intracellular protein that is non-essential for virus replication in cell culture but contributes to virus virulence in vivo and affects the host response to infection. PMID- 16690910 TI - Genotyping Hepatitis B virus from whole- and sub-genomic fragments using position specific scoring matrices in HBV STAR. AB - Hepatitis B virus (HBV) genomes have been classified into eight genotypes based on phylogenetic analysis of sequence variation. Identifying and tracking the movement of HBV genotypes is important in terms of both monitoring infection rates and predicting disease and treatment. An HBV genotyping tool has been developed that compares query sequences with position-specific scoring matrices representing the eight HBV genotypes. This tool (hbv star) is rapid, robust and accurate and assigns genotype based on a statistically defined scoring model. hbv star confidently assigned 90% of 590 full-length HBV genomes to an HBV genotype (Z score >2.0). Thirty-two of the residual 48 sequences were identified as non human primate viruses and 16 sequences were identified as recombinant or putative recombinants. Receiver-Operated Characteristic (ROC) analysis was used to compare the accuracy of genotype prediction using basal core promoter sequences and surface and core genes with the accuracy achieved by using full-length sequences. A web interface to hbv star is available at http://www.vgb.ucl.ac.uk/starn.shtml. PMID- 16690911 TI - Murine gammaherpesvirus-68 glycoprotein H-glycoprotein L complex is a major target for neutralizing monoclonal antibodies. AB - Herpesviruses characteristically persist in immune hosts as latent genomes, but to transmit infection they must reactivate and replicate lytically. The interaction between newly formed virions and pre-existing antibody is therefore likely to be a crucial determinant of viral fitness. Murine gammaherpesvirus-68 (MHV-68) behaves as a natural pathogen of conventional, inbred mice and consequently allows such interactions to be analysed experimentally in a relatively realistic setting. Here, monoclonal antibodies (mAbs) were derived from MHV-68-infected mice and all those recognizing infected-cell surfaces were tested for their capacity to neutralize MHV-68 virions. All of the neutralizing mAbs identified were specific for the viral glycoprotein H (gH)-gL heterodimer and required both gH and gL to reproduce their cognate epitopes. Based on antibody interference, there appeared to be two major neutralization epitopes on gH-gL. Analysis of a representative mAb indicated that it blocked infection at a post-binding step--either virion endocytosis or membrane fusion. PMID- 16690912 TI - Genomic comparison of Neodiprion sertifer and Neodiprion lecontei nucleopolyhedroviruses and identification of potential hymenopteran baculovirus specific open reading frames. AB - Genomic comparison of Neodiprion sertifer nucleopolyhedrovirus (NeseNPV) and Neodiprion lecontei nucleopolyhedrovirus (NeleNPV) showed that the hymenopteran baculoviruses had features in common and were distinct from other, fully sequenced lepidopteran and dipteran baculoviruses. Their genomes were small in size (86,462 and 81,755 bp, respectively), had low G+C contents (33.8 and 33.3 mol%, respectively) and contained fewer open reading frames (ORFs) (90 and 89, respectively) than other baculoviruses. They shared 69 ORFs (48.6% mean amino acid identity overall), 43 of which were previously identified baculovirus homologues. The remaining shared ORFs could be common to other baculoviruses, but low amino acid identities precluded identifying them as such. Some may also be unique to hymenopteran baculoviruses. These included a trypsin-like protease, a zinc-finger protein, regulator of chromosome condensation proteins, a densovirus capsid-like protein and a phosphotransferase. Structural analysis, the presence of conserved domains and phylogenetic studies suggested that some of these ORFs may be functional and could have been transferred horizontally from an insect host. ORFs found only in NeseNPV and NeleNPV may play a role in host specificity and/or tissue tropism, as hymenopteran baculoviruses are restricted to the midgut. The genomes were basically collinear, but contained non-syntenic regions (NSRs) with large numbers of repeats between their polyhedrin and dbp genes. They differed from each other in the number of ORFs and the G+C content of their NSRs and the presence of homologous regions in the NeseNPV genome. NeleNPV also had a short inversion relative to NeseNPV. NeseNPV contained 21 ORFs not found in NeleNPV and NeleNPV had 20 ORFs not found in NeseNPV. PMID- 16690913 TI - In vivo characterization of a group II nucleopolyhedrovirus isolated from Mamestra brassicae (Lepidoptera: Noctuidae) in Japan. AB - A Japanese isolate of Mamestra brassicae nucleopolyhedrovirus (MabrNPV) was identified phylogenetically as a group II nucleopolyhedrovirus (NPV) that is related closely to other NPVs isolated from Mamestra spp. based on nucleotide sequence data of its polh, egt and lef-3 genes. The multiplication of MabrNPV in M. brassicae larvae was characterized following inoculation at various doses and in combination with the fluorescent brightener Tinopal by measuring temporal changes in the concentrations of its viral DNA using real-time quantitative PCR. The growth curves of budded-virus replication were analysed by fitting the data of viral DNA concentration in the host haemolymph to a modified Gompertz model. When fifth-instar larvae were inoculated with an LD95 equivalent dose of MabrNPV and Tinopal, the time lag between the onset of primary and secondary infection was estimated to be 25 h. Another 65 h was required to reach a plateau titre equivalent to a level of 10(9) virions ml(-1) in the haemolymph. All larvae died during the sixth instar following this inoculation regime. In contrast, following inoculation with a 1000-fold higher dose of MabrNPV and Tinopal, the time lag between the onset of primary and secondary infection was only 20 h. Subsequently, the same plateau titre was reached after a further 20 h. Following this inoculation regime, most larvae died during the fifth instar. Quantification of viral DNA by real-time quantitative PCR and application of the Gompertz model are valuable for the characterization of baculovirus replication in vivo. PMID- 16690914 TI - Simian varicella virus gene 28 and 29 promoters share a common upstream stimulatory factor-binding site and are induced by IE62 transactivation. AB - Simian varicella virus (SVV) is a neurotropic alphaherpesvirus that causes a natural, varicella-like disease in non-human primates. After resolution of the primary disease, SVV, like its human counterpart, varicella-zoster virus (VZV), establishes latent infection in the neural ganglia of the host. In this study, gene expression of SVV open reading frames (ORFs) 28 and 29, which encode the viral DNA polymerase and DNA-binding protein, respectively, was characterized during lytic infection of Vero cells. The results indicate that the intergenic region controlling gene 28 and 29 expression includes overlapping, divergent promoters. The ORF 28 and 29 promoters are active in SVV-infected Vero cells, but not in uninfected cells. The SVV immediate-early gene 62 (IE62) product transactivates ORF 28 and 29 expression, and a cellular upstream stimulatory factor-binding site is important for efficient IE62 induction of genes 28 and 29. DNA sequence analysis of the 185 bp intergenic region identified putative cellular transcription factor-binding sites. Transcriptional analysis mapped ORF 28 and 29 RNA start sites. A recombinant SVV was employed to demonstrate that the ORF 29 promoter can express a heterologous gene (green fluorescent protein) when inserted into a novel site (the ORF 12/13 intergenic region) within the SVV genome. The findings demonstrate similarities between SVV and VZV ORF 28/29 expression and indicate that the simian varicella model may be useful to investigate the differential regulation of viral genes during lytic and latent infection. PMID- 16690915 TI - Evolution of Bovine herpesvirus 4: recombination and transmission between African buffalo and cattle. AB - Bovine herpesvirus 4 (BoHV-4) has been isolated from cattle throughout the world, but virological and serological studies have suggested that the African buffalo is also a natural host for this virus. It has previously been found that the Bo17 gene of BoHV-4 was acquired from an ancestor of the African buffalo, probably around 1.5 million years ago. Analysis of the variation of the Bo17 gene sequence among BoHV-4 strains suggested a relatively ancient transmission of BoHV-4 from the buffalo to the Bos primigenius lineage, followed by a host-dependent split between zebu and taurine BoHV-4 strains. In the present study, the evolutionary history of BoHV-4 was investigated by analysis of five gene sequences from each of nine strains representative of the viral species: three isolated from African buffalo in Kenya and six from cattle from Europe, North America and India. No two gene sequences had the same evolutionary tree, indicating that recombination has occurred between divergent lineages; six recombination events were delineated for these sequences. Nevertheless, exchange has been infrequent enough that a clonal evolutionary history of the strains could be discerned, upon which the recombination events were superimposed. The dates of divergence among BoHV-4 lineages were estimated from synonymous nucleotide-substitution rates. The inferred evolutionary history suggests that African buffalo were the original natural reservoir of BoHV-4 and that there have been at least three independent transmissions from buffalo to cattle, probably via intermediate hosts and--at least in the case of North American strains--within the last 500 years. PMID- 16690916 TI - Vaccinia virus kelch protein A55 is a 64 kDa intracellular factor that affects virus-induced cytopathic effect and the outcome of infection in a murine intradermal model. AB - The vaccinia virus (VACV) protein A55 is a BTB/kelch protein with a broad complex, tramtrack and bric-a-brac (BTB) domain in the N-terminal region and five kelch repeats in the C-terminal half. The BTB/kelch subgroup of the kelch superfamily of proteins has been associated with a wide variety of functions including regulation of the cytoskeleton. VACV contains three genes predicted to encode BTB/kelch proteins: A55R, F3L and C2L. The A55R gene product has been identified as an intracellular protein of 64 kDa that is expressed late in infection. A VACV strain lacking 93.6% of the A55R open reading frame (vdeltaA55) was constructed and found to have an unaltered growth rate in vivo but a different plaque morphology and cytopathic effect, as well as reduced development of VACV-induced Ca2+-independent cell/extracellular matrix adhesion. In a murine intradermal model of VACV infection, a virus lacking the A55R gene induced larger lesions than wild-type and revertant control viruses. PMID- 16690917 TI - Comparative genomic analysis of two strains of human adenovirus type 3 isolated from children with acute respiratory infection in southern China. AB - Human adenovirus type 3 (HAdV-3) is a causative agent of acute respiratory disease, which is prevalent throughout the world, especially in Asia. Here, the complete genome sequences of two field strains of HAdV-3 (strains GZ1 and GZ2) isolated from children with acute respiratory infection in southern China are reported (GenBank accession nos DQ099432 and DQ105654, respectively). The genomes were 35,273 bp (GZ1) and 35,269 bp (GZ2) and both had a G+C content of 51 mol%. They shared 99% nucleotide identity and the four early and five late regions that are characteristic of human adenoviruses. Thirty-nine protein- and two RNA-coding sequences were identified in the genome sequences of both strains. Protein pX had a predicted molecular mass of 8.3 kDa in strain GZ1; this was lower (7.6 kDa) in strain GZ2. Both strains contained 10 short inverted repeats, in addition to their inverted terminal repeats (111 bp). Comparative whole-genome analysis revealed 93 mismatches and four insertions/deletions between the two strains. Strain GZ1 infection produced a typical cytopathic effect, whereas strain GZ2 did not; non-synonymous substitutions in proteins of GZ2 may be responsible for this difference. PMID- 16690918 TI - Identification of novel murine parvovirus strains by epidemiological analysis of naturally infected mice. AB - Random-source DNA samples obtained from naturally infected laboratory mice (n=381) were evaluated by PCR and RFLP analysis to determine the prevalence of murine parvovirus strains circulating in contemporary laboratory mouse colonies. Mouse parvovirus (MPV) was detected in 77% of samples, Minute virus of mice (MVM) was detected in 16% of samples and both MVM and MPV were detected in 7% of samples. MVMm, a strain recently isolated from clinically ill NOD-mu chain knockout mice, was detected in 91% of MVM-positive samples, with the Cutter strain of MVM (MVMc) detected in the remaining samples. The prototypic and immunosuppressive strains of MVM were not detected in any of the samples. MPV-1 was detected in 78% of the MPV-positive samples and two newly identified murine parvoviruses, tentatively named MPV-2 and MPV-3, were detected in 21 and 1% of the samples, respectively. The DNA sequence encompassing coding regions of the viral genome and the predicted protein sequences for MVMm, MPV-2 and MPV-3 were determined and compared with those of other rodent parvovirus strains and LuIII parvovirus. The genomic organization for the newly identified viral strains was similar to that of other rodent parvoviruses, and nucleotide sequence identities indicated that MVMm was most similar to MVMc (96.1%), MPV-3 was most similar to hamster parvovirus (HaPV) (98.1%) and MPV-2 was most similar to MPV-1 (95.3%). The genetic similarity of MPV-3 and HaPV suggests that HaPV epizootics in hamsters may result from cross-species transmission, with mice as the natural rodent host for this virus. PMID- 16690919 TI - Activation of early gene transcription in polyomavirus BK by human immunodeficiency virus type 1 Tat. AB - Polyomavirus BK (BKV) is a serious problem for immunocompromised patients, where latent virus can enter into the lytic cycle causing cytolytic destruction of host cells. BKV infects >80% of the population worldwide during childhood and then remains in a latent state in the kidney. In the context of immunosuppression in kidney transplant patients, reactivation of the viral early promoter (BKV(E)) results in production of T antigen, enabling virus replication and transition from latency to the lytic phase, causing polyomavirus-associated nephropathy. Reactivation of BKV can also cause complications such as nephritis, atypical retinitis and haemorrhagic cystitis in AIDS patients. Here, the effects of human immunodeficiency virus type 1 (HIV-1) proteins Tat and Vpr on BKV transcription were investigated and it was demonstrated that Tat dramatically stimulated BKV(E). Site-directed mutagenesis analysis of potential Tat-responsive transcriptional motifs complemented by an electrophoretic mobility shift assay (EMSA) showed that Tat activated BKV(E) by inducing binding of the NF-kappaB p65 subunit to a kappaB motif near the 3' end of BKV(E). In addition, a sequence within the 5' UTR of BKV(E) transcripts (BKV(E)-TAR) was identified that is identical to the HIV-1 transactivation response (TAR) element. The BKV(E)-TAR sequence bound TAT in RNA EMSA assays and deletion of the BKV(E)-TAR sequence eliminated Tat transactivation of BKV(E) transcription. Thus, Tat positively affected BKV(E) transcription by a dual mechanism and this may be important in diseases involving BKV reactivation in AIDS patients. PMID- 16690920 TI - Characterization of a new densovirus infecting the German cockroach, Blattella germanica. AB - A new DNA virus (Parvoviridae: Densovirinae, Densovirus) was isolated and purified from descendants of field-collected German cockroaches, Blattella germanica. Viral DNA and cockroach tissues infected with B. germanica densovirus (BgDNV) were examined by electron microscopy. Virus particles, about 20 nm in diameter, were observed both in the nucleus and in the cytoplasm of infected cells. Virus DNA proved to be a linear molecule of about 1.2 microm in length. BgDNV isolated from infected cockroaches infected successfully and could be maintained in BGE-2, a B. germanica cell line. The complete BgDNV genome was sequenced and analysed. Five open reading frames (ORFs) were detected in the 5335 nt sequence: two ORFS that were on one DNA strand encoded structural capsid proteins (69.7 and 24.8 kDa) and three ORFs that were on the other strand encoded non-structural proteins (60.2, 30.3 and 25.9 kDa). Three putative promoters and polyadenylation signals were identified. Structural analysis of the inverted terminal repeats revealed the presence of extended palindromes. The genome structure of BgDNV was compared with that of other members of the family Parvoviridae; the predicted amino acid sequences were aligned and subjected to phylogenetic analyses. PMID- 16690921 TI - Envelope proteins of spleen necrosis virus form infectious human immunodeficiency virus type 1 pseudotype vector particles, but fail to incorporate upon substitution of the cytoplasmic domain with that of Gibbon ape leukemia virus. AB - The wild-type (wt) envelope (Env) proteins of spleen necrosis virus (SNV), together with the transmembrane (TM) protein fused to antibody domains (scFv), have been used for the generation of stable packaging cell lines releasing pseudotyped cell targeting vectors derived from SNV and Murine leukemia virus (MLV). As a first step towards assessing whether HIV-1(SNV/TM-scFv) packaging cells could be established for the production of lentiviral cell targeting vectors, it is reported here that infectious HIV-1-derived particles pseudotyped with wt SNV Env proteins could be generated. Using novel chimeric SNV-derived Env proteins encompassing wt and engineered cytoplasmic domains (C-tail) of the Gibbon ape leukemia virus (GaLV) TM protein, it was further shown that the wt C tail not only excludes the GaLV TM protein from incorporation into HIV-1 particles, but confers this phenotype to other retroviral envelopes upon C terminal fusion. PMID- 16690922 TI - Murine leukemia virus transmembrane protein R-peptide is found in small virus core-like complexes in cells. AB - The core of the retrovirus Murine leukemia virus (MLV) consists of the Gag precursor protein and viral RNA. It assembles at the cytoplasmic face of the cell membrane where, by an unclear mechanism, it collects viral envelope proteins embedded in the cell membrane and buds off. The C-terminal half of the short cytoplasmic tail of the envelope transmembrane protein (TM) is cleaved off to yield R-peptide and fusion-active TM. In Moloney MLV particles, R-peptide was found to bind to core particles. In cells, R-peptide and low amounts of uncleaved TM were found to be associated with small core-like complexes, i.e. mild detergent-insoluble, Gag-containing complexes with a density of 1.23 g ml(-1) and a size of 150-200 S. Our results suggest that TM associates with the assembling core particle through the R-peptide before budding and that this is the mechanism by which the budding virus acquires the envelope proteins. PMID- 16690923 TI - Suppression of human immunodeficiency virus type 1 replication by arginine deiminase of Mycoplasma arginini. AB - It was found previously that human immunodeficiency virus type 1 (HIV-1) irrelevant CD8+ cytotoxic T lymphocytes (CTLs) from uninfected donors suppressed HIV-1 replication in a cell-contact-dependent manner. However, one of these CTL lines (CTL-3) also significantly suppressed HIV-1 replication through its supernatant. Here, the suppressive fraction from CTL-3 supernatant was purified and analysed by mass spectrometry. A protein band specific for the suppressive fraction was identified as arginine deiminase from Mycoplasma arginini, which catalyses the hydrolysis of arginine to citrulline. Addition of L-arginine or the use of antibiotics against mycoplasma restored supernatant-mediated but not cell contact-dependent suppression of HIV-1 replication by CTL-3, clearly indicating that arginine deiminase of M. arginini in the supernatants suppressed HIV-1 replication, which is independent of CD8+ T-cell-mediated HIV-1 suppression via cell contact. Arginine deiminase is known to be a chemotherapeutic agent against arginine-requiring tumours and these results suggest that it also has potential application in antiviral therapy. PMID- 16690924 TI - Quantifying antagonistic epistasis in a multifunctional RNA secondary structure of the Rous sarcoma virus. AB - Recent studies have suggested that antagonistic epistasis (i.e. mutations having smaller effects in combination than alone) may be common among RNA viruses, in contrast to other biological systems. Here, by re-analysing previously published data from a random viral library, selection and epistasis coefficients were estimated in the U5-IR stem and loop of the Rous sarcoma virus, a region that adopts a conserved secondary structure and is involved in various essential steps of viral infection. The estimated mutational fitness effects are extremely high and genetic interactions are antagonistic on average. This pattern might be representative of RNA virus genomes, which show high compaction and frequent secondary structures. The implications for RNA virus adaptability are explored. PMID- 16690925 TI - Human immunodeficiency virus type 1 Tat increases cooperation between AP-1 and NFAT transcription factors in T cells. AB - Human immunodeficiency virus type 1 (HIV-1) Tat affects cellular gene expression through modulation of the activity of different transcription factors. Here, the role of Tat in the cooperation between nuclear factor of activated T cells (NFAT) and activator protein 1 (AP-1) transcription factors was investigated. Constitutive or transient Tat expression in Jurkat T cells enhanced cooperative NFAT/AP-1- but not AP-1-dependent transcription independent of its ability to transactivate the HIV-1 LTR. The enhancing effect of Tat took place after nuclear translocation of NFAT. Furthermore, transactivation of an NFAT/AP-1 reporter by transfection of NFAT and c-Jun was strongly enhanced by simultaneous Tat transfection. Moreover, intracellular Tat expression increased the binding of NFAT/AP-1 complexes to the interleukin 2 promoter without significantly altering NFAT- and AP-1-independent binding. HIV-1 Tat interacted with NFAT but not c-Jun. These results indicate that Tat interacts with NFAT, affecting its cooperation with AP-1, without altering independent binding of these transcription factors to DNA. PMID- 16690926 TI - Human immunodeficiency virus type 1 Tat prevents dephosphorylation of Sp1 by TCF 4 in astrocytes. AB - Previous examination of the effect of TCF-4 on transcription of the human immunodeficiency virus type 1 (HIV-1) promoter in human astrocytic cells found that TCF-4 affects the HIV-1 promoter through the GC-rich domain (nt -80 to nt 68). Here, the physical interaction and a functional consequence of TCF4-Sp1 contact were characterized. It was shown that expression of TCF-4 in U-87 MG (human astrocytic) cells decreased basal and Sp1-mediated transcription of the HIV-1 promoter. Results from a GST pull-down assay, as well as combined immunoprecipitation and Western blot analysis of protein extracts from U-87 MG cells, revealed an interaction of Sp1 with TCF-4. Using in vitro protein chromatography, the region of Sp1 that contacts TCF-4 was mapped to aa 266-350. It was also found that, in cell-free extracts, TCF-4 prevented dsDNA-dependent protein kinase (DNA-PK)-mediated Sp1 phosphorylation. Surprisingly, TCF-4 failed to decrease Sp1-mediated transcription of the HIV-1 long terminal repeat (LTR) and Sp1 phosphorylation in cells expressing HIV-1 Tat. Results from immunoprecipitation/Western blotting demonstrated that TCF-4 lost its ability to interact with Sp1, but not with Tat, in Tat-transfected cells. Taken together, these findings suggest that activity at the HIV-1 promoter is influenced by phosphorylation of Sp1, which is affected by Tat and DNA-PK. Interactions among TCF-4, Sp1 and/or Tat may determine the level of viral gene transcription in human astrocytic cells. PMID- 16690927 TI - A single administration of lentiviral vectors expressing either full-length human immunodeficiency virus 1 (HIV-1)(HXB2) Rev/Env or codon-optimized HIV-1(JR-FL) gp120 generates durable immune responses in mice. AB - Genetic immunization using viral vectors provides an effective means to elicit antigen-specific cellular immune responses. Several viral vectors have proven efficacious in inducing immune responses after direct injection in vivo. Among them, recombinant, self-inactivating lentiviral vectors are very attractive delivery systems, as they are able to efficiently transduce into and express foreign genes in a wide variety of mammalian cells. A self-inactivating lentiviral vector was evaluated for the delivery of human immunodeficiency virus 1 (HIV-1) envelope sequences in mice in order to elicit specific immune responses. With this aim, BALB/c mice were immunized with a single injection of self-inactivating lentiviral vectors carrying either the full-length HIV-1(HXB2) Rev/Env (TY2-IIIBEnv) or the codon-optimized HIV-1(JR-FL) gp120 (TY2-JREnv) coding sequence. Both vectors were able to elicit specific cellular responses efficiently, as measured by gamma interferon ELISPOT and chromium-release assays, upon in vitro stimulation of splenocytes from BALB/c immunized mice. However, only the TY2-JREnv-immunized mice were able to elicit specific humoral responses, measured as anti-gp120 antibody production. These data provide the first evidence that a single, direct, in vivo administration of a lentiviral vector encoding a viral gene might represent a useful strategy for vaccine development. PMID- 16690928 TI - CD9-dependent regulation of Canine distemper virus-induced cell-cell fusion segregates with the extracellular domain of the haemagglutinin. AB - Antibodies to CD9, a member of the tetraspan transmembrane-protein family, selectively inhibit Canine distemper virus (CDV)-induced cell-cell fusion. Neither CDV-induced virus-cell fusion nor cell-cell fusion induced by the closely related morbillivirus Measles virus (MV) is affected by anti-CD9 antibodies. As CDV does not bind CD9, an unknown, indirect mechanism is responsible for the observed inhibition of cell-cell fusion. It was investigated whether this effect was restricted to only one viral glycoprotein, either the haemagglutinin (H) or the fusion (F) protein, which form a fusion complex on the surface of virions and infected cells, or whether it is dependent on both in transient co-transfection assays. The susceptibility to CD9 antibodies segregates with the H protein of CDV. By exchanging portions of the H proteins of CDV and MV, it was determined that the complete extracellular domain, including the predicted stem structure (stem 1, barrel strand 1 and stem 2) and globular head domain, of the CDV-H protein mediates the effect. This suggests that interaction of the CDV-H protein with an unknown cellular receptor(s) is regulated by CD9, rather than F protein mediated membrane fusion. PMID- 16690929 TI - Recombinant wild-type measles virus containing a single N481Y substitution in its haemagglutinin cannot use receptor CD46 as efficiently as that having the haemagglutinin of the Edmonston laboratory strain. AB - Signalling lymphocyte activation molecule (SLAM) acts as a cellular receptor for Measles virus (MV). The recombinant MV, based on a SLAM-using clinical isolate in which asparagine at position 481 of the haemagglutinin was replaced with tyrosine, was generated. Characterization of this recombinant virus revealed that the N481Y substitution in the haemagglutinin allowed it to utilize CD46 as an alternative receptor, but that its ability to use CD46 was rather low in CD46+ SLAM- cell lines compared with that of the recombinant virus possessing the haemagglutinin of the Edmonston laboratory strain. Thus, an N481Y substitution alone may not be sufficient to make SLAM-using MVs use CD46 efficiently, suggesting that further substitutions in the haemagglutinin are required for them to grow efficiently in CD46+ cells like the Edmonston strain. This may be a reason why few CD46-using MVs are detected in vivo. PMID- 16690930 TI - Sequence elements of the fusion peptide of human respiratory syncytial virus fusion protein required for activity. AB - We have reported previously the expression and purification of an anchorless form of the human respiratory syncytial virus (HRSV) F protein (F(TM-)) representing the ectodomain of the full-length F. F(TM-) molecules are seen as unaggregated cones by electron microscopy but completion of proteolytic cleavage of the F0 monomers in the F(TM-) trimer leads to a change in shape from cones to lollipops that aggregate into rosettes. This aggregation apparently occurs by interaction of the fusion peptides of F(TM-) molecules that are exposed after cleavage. Since exposure of the fusion peptide is a key event in the process of membrane fusion, changes associated with F(TM-) cleavage may reflect those occurring in full length F during membrane fusion. Deletions or substitutions that changed either the length, charge or hydrophobicity of the fusion peptide inhibited aggregation of F(TM-), and these mutants remained as unaggregated cones after cleavage. In contrast, more conservative changes did not inhibit the change of shape and aggregation of F(TM-). When the same changes were introduced in the fusion peptide of full-length F, only the mutations that inhibited aggregation of F(TM-) prevented membrane fusion. Thus, the conformational changes that follow completion of cleavage of the F(TM-) protein require a functional fusion peptide. These sequence constraints may restrict accumulation of sequence changes in the fusion peptide of HRSV F when compared with other hydrophobic regions of the molecule. PMID- 16690931 TI - Bovine respiratory syncytial virus lacking the virokinin or with a mutation in furin cleavage site RA(R/K)R109 induces less pulmonary inflammation without impeding the induction of protective immunity in calves. AB - The BRSV fusion (F) protein is cleaved at two furin consensus sequence sites, resulting in the generation of disulphide-linked F1 and F2 subunits and the release of an intervening peptide of 27 amino acids (pep27), which is converted into a biologically active tachykinin (virokinin). The role of the virokinin and the importance of one of the furin cleavage sites, FCS-2 [RA(R/K)R109], in the pathogenesis of BRSV infection and in the subsequent development of immunity was studied in gnotobiotic calves infected with a recombinant BRSV (rBRSV) lacking pep27 (rBRSVdelta p27) or with rBRSV108/109, which contains two amino acid substitutions in FCS-2 (RANN109). Although replication of the mutant viruses and the parental wild-type (WT) rBRSV in the lungs was similar, the extent of gross and microscopic lesions induced by the mutant viruses was less than that induced by WT rBRSV. Furthermore, the numbers of eosinophils in the lungs of calves infected with the mutant viruses were significantly less than that in calves infected with WT virus. These observations suggest a role for the virokinin in the pathogenesis of BRSV infection. Following mucosal immunization with rBRSVdelta p27, the levels of BRSV-specific serum antibodies were similar to those induced by WT virus. In contrast, the level of neutralizing antibodies induced by rBRSV108/109 was 10-fold lower than that induced by WT virus. Nevertheless, resistance to BRSV challenge induced by the mutant and WT viruses was similar, suggesting that neither pep27 nor FCS-2 plays a major role in the induction of protective immunity. PMID- 16690932 TI - A point mutation at the C terminus of the cytoplasmic domain of influenza B virus haemagglutinin inhibits syncytium formation. AB - The C-terminal sequence of the cytoplasmic tail (CT) of influenza B haemagglutinin (BHA) consists of strictly conserved, hydrophobic amino acids, and the endmost C-terminal amino acid of the CT is Leu. To elucidate the role of this amino acid in the fusion activity of BHA (B/Kanagawa/73), site-specific mutant HAs were created by replacing Leu at this position with Arg, Lys, Ser, Try, Val or Ile or by the deletion of Leu altogether. All mutants were expressed at the cell surface, bound to red blood cells, were cleaved properly into two subunits and could be acylated like the wild-type (wt) HA. The membrane-fusion ability of these mutants was examined with a lipid (R18) and aqueous (calcein) dye-transfer assay and quantified with a syncytium-formation assay. All mutant HAs showed no measurable effect on lipid mixing or fusion-pore formation. However, mutant HAs with a hydrophobic value of the C-terminal amino acid lower than that of Leu had a reduced ability to form syncytia, whereas mutants with a more hydrophobic amino acid (Val or Ile) promoted fusion to the extent of the wt HA. On the other hand, the mutant HA with the deletion of Leu supported full fusion. These results demonstrate that Leu at the endmost portion of the C terminus of the BHA-CT is not essential for BHA-mediated fusion, but that the hydrophobicity of the single amino acid at this position plays an important role in syncytium formation. PMID- 16690933 TI - Analysis of gene-expression profiles by oligonucleotide microarray in children with influenza. AB - In order to clarify the mechanism of the host response to influenza virus, gene expression profiles of peripheral blood obtained from paediatric patients with influenza were investigated by oligonucleotide microarray. In the acute phase of influenza, 200 genes were upregulated and 20 genes were downregulated compared with their expression in the convalescent phase. Interferon-regulated genes, such as interferon-induced protein with tetratricopeptide repeats 2 (IFIT2) and vipirin, were strongly upregulated in the acute phase. Gene ontology analysis showed that immune response genes were highly overrepresented among the upregulated genes. Gene-expression profiles of influenza patients with and without febrile convulsion were also studied. In patients with febrile convulsion, 22 genes were upregulated and five were downregulated compared with their expression in patients without febrile convulsion. These results should help to clarify the pathogenesis of influenza and its neurological complications. PMID- 16690934 TI - Feline infectious peritonitis virus-infected monocytes internalize viral membrane bound proteins upon antibody addition. AB - Feline infectious peritonitis virus (FIPV) may cause a highly lethal infection in cats, in spite of a usually strong humoral immune response. Antibodies seem unable to identify infected cells and mediate antibody-dependent cell lysis. In this study, the effect of antibodies on Feline coronavirus (FCoV)-infected monocytes was investigated. Upon addition of FCoV-specific antibodies, surface expressed viral proteins were internalized through a highly efficient process, resulting in cells without visually detectable viral proteins on their plasma membrane. The internalization was also induced by mAbs against the Spike and Membrane proteins, suggesting that both proteins play a role in the process. The internalization did not occur spontaneously, as it was not observed in cells incubated with medium or non-specific antibodies. Further, the internalization could not be reproduced in feline cell lines, indicating its cell-type specificity. This study sheds new light on the immune-evasive nature of FIPV infections. PMID- 16690935 TI - Analysis of ACE2 in polarized epithelial cells: surface expression and function as receptor for severe acute respiratory syndrome-associated coronavirus. AB - The primary target of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) is epithelial cells in the respiratory and intestinal tract. The cellular receptor for SARS-CoV, angiotensin-converting enzyme 2 (ACE2), has been shown to be localized on the apical plasma membrane of polarized respiratory epithelial cells and to mediate infection from the apical side of these cells. Here, these results were confirmed and extended by including a colon carcinoma cell line (Caco-2), a lung carcinoma cell line (Calu-3) and Vero E6 cells in our analysis. All three cell types expressed human ACE2 on the apical membrane domain and were infected via this route, as determined with vesicular stomatitis virus pseudotypes containing the S protein of SARS-CoV. In a histological analysis of the respiratory tract, ACE2 was detected in the trachea, main bronchus and alveoli, and occasionally also in the small bronchi. These data will help us to understand the pathogenesis of SARS-CoV infection. PMID- 16690936 TI - A new subtype of hepatitis C virus genotype 1: complete genome and phylogenetic relationships of an Equatorial Guinea isolate. AB - Hepatitis C virus (HCV) is the leading cause of chronic liver disease and is associated with hepatocellular carcinoma. However, there have been few studies on the distribution and genetic diversity of HCV isolates in non-developed countries. Here, the complete genome sequence of an HCV genotype 1 isolate from Equatorial Guinea is reported, the first complete HCV-1 genome of African origin. Phylogenetic analysis revealed that this sequence always grouped with sequences of genotype 1, but did not group clearly with any subtype described so far. An analysis of partial NS5B gene sequences with additional sequences of African origin also failed to find close similarities between the new sequence and any previously known isolate. Genetic divergence of the coding region of this new sequence with respect to the recognized subtypes of HCV-1 ranged from 20 to 22%. It is proposed that this isolate is a representative of a new, distinct variant of HCV subtype 1. PMID- 16690937 TI - NS3 protein of Hepatitis C virus associates with the tumour suppressor p53 and inhibits its function in an NS3 sequence-dependent manner. AB - The N-terminal 198 residues of NS3 (NS3-N) of Hepatitis C virus (HCV) subtype 1b obtained from 29 patients, as well as full-length NS3 (NS3-Full), were analysed for their subcellular localization, interaction with the tumour suppressor p53 and serine protease activity in the presence and absence of the viral cofactor NS4A. Based on the subcellular-localization patterns in the absence of NS4A, NS3 N sequences were classified into three groups, with each group exhibiting either dot-like, diffuse or a mixed type of localization. Chimeric NS3-Full sequences, each consisting of an individual NS3-N and a shared C-terminal sequence, showed the same localization patterns as those of the respective NS3-N. Site-directed mutagenesis experiments revealed that a single or a few amino acid substitutions at a particular position(s) of NS3-N altered the localization pattern. Interestingly, NS3 of the dot-like type, either NS3-N or NS3-Full, interacted with p53 more strongly than that of the diffuse type, in both the presence and the absence of NS4A. Moreover, NS3-N of the dot-like type suppressed trans activating activity of p53 more strongly than that of the diffuse type. Serine protease activity did not differ significantly between the two types of NS3. In HCV RNA replicon-harbouring cells, physical interaction between NS3 and p53 was observed consistently and p53-mediated transcriptional activation was suppressed significantly compared with HCV RNA-negative control cells. Our results collectively suggest the possibility that NS3 plays an important role in the hepatocarcinogenesis of HCV by interacting differentially with p53 in an NS3 sequence-dependent manner. PMID- 16690938 TI - Induction of hepatitis D virus large antigen translocation to the cytoplasm by hepatitis B virus surface antigens correlates with endoplasmic reticulum stress and NF-kappaB activation. AB - It is known that hepatitis D virus (HDV) requires hepatitis B virus (HBV) for supplying envelope proteins (HBsAgs) to produce mature virions, and the HDV large antigen (LDAg) is responsible for interacting with HBsAgs. However, the signal molecules involved in the cross-talk between HBsAgs and LDAg have never been reported. It has been previously demonstrated that the small form of HBsAg can facilitate the translocation of HDV large antigen green fluorescent protein (GFP) fusion protein (GFP-LD) from the nucleus to the cytoplasm. In this study, it was confirmed that the small form of HBsAg can facilitate both GFP-LD and authentic LDAg for nuclear export. It was also shown that the three forms of HBsAgs (large, middle and small) induced various rates (from 35.4 to 57.2%) of GFP-LD nuclear export. Since HBsAgs are localized inside the endoplasmic reticulum (ER), this suggests that ER stress possibly initiates the signal for inducing LDAg translocation. This supposition is supported by results that show that around 9% of cells appear with GFP-LD in the cytoplasm after treatment with the ER stress inducers, brefeldin A (BFA) and tunicamycin, in the absence of HBsAg. Western blot and immunofluorescence microscopy results further showed that the activation of NF-kappaB is linked to the ER stress that induces GFP-LD translocation. Combining this with results showing that tumour necrosis factor alpha (TNF-alpha) can also induce GFP-LD translocation, it was concluded that LDAg translocation correlates with ER stress and activation of NF-kappaB. Nevertheless, TNF-alpha induced GFP-LD translocation was independent of new protein synthesis, suggesting that a post-translational event occurs to GFP-LD to allow translocation. PMID- 16690939 TI - Genetic diversity analyses of grapevine Rupestris stem pitting-associated virus reveal distinct population structures in scion versus rootstock varieties. AB - Grapevine Rupestris stem pitting-associated virus (GRSPaV) is a member of the genus Foveavirus within the family Flexiviridae. GRSPaV is closely associated with the disease Rupestris stem pitting and is frequently detected in grapevines worldwide. Previous research in several laboratories suggests that GRSPaV consists of a family of sequence variants. However, the genetic composition of GRSPaV variants in viral isolates from scion and rootstock varieties has not been studied extensively. In this report, the genetic diversity and population structure of GRSPaV isolates from scion and rootstock varieties were analysed using two pairs of primers targeting two different genomic regions encoding the helicase domain of the replicase and the capsid protein. In total, 190 cDNA clones derived from 24 isolates were sequenced and analysed. At least four major groups of GRSPaV variants were found to exist in grapevines. Interestingly, the majority of the scion varieties (9/10) that were analysed, regardless of their genetic background and geographical origin, harboured complex viral populations composed of two to four distinct viral variants. In contrast, the viral populations in isolates from rootstock varieties were homogeneous and comprised a single variant. The practice of grafting between scion and rootstock varieties commonly used in modern viticulture, coupled with the frequent regional and international exchange of propagating materials, may have played a major role in the ubiquitous distribution and mixed infections of distinct GRSPaV variants among scion varieties. The possible origin and evolution of GRSPaV are also discussed. PMID- 16690940 TI - Infectivity of nanovirus DNAs: induction of disease by cloned genome components of Faba bean necrotic yellows virus. AB - Circumstantial evidence suggests that the genome of Faba bean necrotic yellows virus (FBNYV), a nanovirus, consists of eight distinct, circular, single-stranded DNAs, each of about 1 kb and encoding only one protein. Here, the use of cloned full-length FBNYV DNAs for reproducing FBNYV-like symptoms in Vicia faba, the principal natural host of FBNYV, is reported. Characteristic symptoms of FBNYV infection were obtained in faba bean plants following biolistic DNA delivery or agroinoculation with all eight FBNYV DNAs. Although the eight different DNAs have been invariably detected in field samples infected with the various geographical FBNYV isolates, experimental infection with different combinations of fewer than eight DNAs also led to typical FBNYV symptoms. Even only five genome components, DNA-R, DNA-S, DNA-M, DNA-U1 and DNA-U2, were sufficient for inducing disease symptoms in V. faba upon agroinoculation. Symptomatic plants agroinoculated or bombarded with eight DNAs contained typical FBNYV virions; however, the virus was not transmitted by Aphis craccivora or Acyrthosiphon pisum, two efficient aphid vectors of FBNYV. PMID- 16690941 TI - In vitro and in vivo mapping of the Prunus necrotic ringspot virus coat protein C terminal dimerization domain by bimolecular fluorescence complementation. AB - Interactions between viral proteins are critical for virus viability. Bimolecular fluorescent complementation (BiFC) technique determines protein interactions in real-time under almost normal physiological conditions. The coat protein (CP) of Prunus necrotic ringspot virus is required for multiple functions in its replication cycle. In this study, the region involved in CP dimerization has been mapped by BiFC in both bacteria and plant tissue. Full-length and C-terminal deleted forms of the CP gene were fused in-frame to the N- and C-terminal fragments of the yellow fluorescent protein. The BiFC analysis showed that a domain located between residues 9 and 27 from the C-end plays a critical role in dimerization. The importance of this C-terminal region in dimer formation and the applicability of the BiFC technique to analyse viral protein interactions are discussed. PMID- 16690942 TI - Characterization of proteinase K-resistant N- and C-terminally truncated PrP in Nor98 atypical scrapie. AB - An increasing number of scrapie cases with atypical characteristics, designated Nor98, have recently been recognized. Here, the proteinase K (PK)-resistant prion protein (PrP) fragments from two Swedish cases of Nor98 atypical scrapie have been characterized. The prominent, fast-migrating band in the distinct Nor98 Western immunoblot electrophoretic profile was determined to be of 7 kDa in size and was accordingly designated Nor98-PrP7. The antigenic composition of Nor98 PrP7, as assayed by a panel of anti-PrP antibodies, revealed that this fragment comprised a mid-region of PrP from around aa 85 to 148. N- and C-terminally truncated fragments spanning the mid-region of PrP have only been observed in the genetic prion disorder Gerstmann-Straussler-Scheinker disease. It is shown here that the long-term PK resistance of Nor98-PrP7 is reduced compared with that of PrP(res) in classical scrapie. Enzymic deglycosylation did not change the distinct electrophoretic profile of Nor98-PrP7. A previously unidentified, PK resistant, C-terminal PrP fragment of around 24 kDa was detected and its PK resistance was investigated. After deglycosylation, this fragment migrated as a 14 kDa polypeptide and was designated PrP-CTF14. Antigenic determination and the size of 14 kDa suggested a fragment spanning approximately aa 120-233. The existence of two PK-resistant PrP fragments, Nor98-PrP7 and PrP-CTF14, that share an overlapping region suggests that at least two distinct PrP conformers with different PK-resistant cores are present in brain extracts from Nor98-affected sheep. The structural gene of PrP in three Nor98-affected sheep was analysed, but no mutations were found that could be correlated to the aberrant PK-resistant profile observed. PMID- 16690944 TI - Reoxygenation with 100% oxygen versus room air: late neuroanatomical and neurofunctional outcome in neonatal mice with hypoxic-ischemic brain injury. AB - Study investigated neuroutcome in mice subjected at 7-8 d of life to hypoxic ischemic brain injury (HI) followed by 30 min of reoxygenation with 100% O(2) (Re O(2)) or room air (Re-Air). At 24 h of recovery, mouse reflexes were tested. At 7 wks after HI spatial orientation and memory were assessed in the same mice. Mortality rate was recorded at 24 h and at 7 wks of recovery. In separate cohort of mice, changes in cerebral blood flow (CBF) during HI-insult and reoxygenation were recorded. Re-O(2)versus Re-Air mice exhibited significantly delayed geotaxis reflex. Adult Re-O(2)versus Re-Air mice exhibited significantly better spatial learning and orientation with strong tendency toward better preserved memory. Histopathology revealed significantly less hippocampal atrophy in Re-O(2)versus Re-Air mice. Following a hypoxia-induced hypoperfusion, Re-O(2) re-established CBF in the ipsilateral side to the prehypoxic level significantly faster than Re Air. The mortality was higher among Re-O2 versus Re-Air mice, although, it did not reach statistical significance. Re-O(2)versus Re-Air restores CBF significantly faster and results in better late neuroutcome. However, greater early motor deficit and higher mortality rate among Re-O(2)versus Re-Air mice suggest that Re-O(2) may be deleterious at the early stage of recovery. PMID- 16690945 TI - Effect of maternal protein restriction in rats on cardiac fibrosis and capillarization in adulthood. AB - This study examines the effect of maternal protein restriction in rats on levels of cardiac fibrosis, myocardial capillarization, and media:lumen ratio of intramyocardial arteries in adult offspring. Female Wistar Kyoto rats were fed either a normal protein diet (NPD; 20% casein) or a low-protein diet (LPD; 8.7% casein) during pregnancy and lactation. Female offspring (seven per group) were weaned at 4 wk of age and grown to adulthood. At 24 wk of age, the offspring were perfusion fixed. Cardiac fibrosis and media:lumen ratio of intramyocardial arterioles was assessed using image analysis and cardiac capillarization was stereologically investigated. Body weights at 2 and 24 wk of age were significantly reduced (31% and 8%, respectively) in the LPD offspring; however, heart size was not different at 24 wk. Importantly by adulthood, there was a significant 15% increase in left ventricular interstitial fibrosis in LPD offspring. There were no differences in levels of perivascular fibrosis, myocardial capillarization, or in the media:lumen ratio of intramyocardial arteries between groups. Because cardiac fibrosis is associated with impaired cardiac contractility and arrhythmia, our results suggest that induction of interstitial fibrosis may contribute to the increased cardiac disease in adult subjects who were exposed to an adverse intrauterine environment. PMID- 16690946 TI - Prenatal estrogen and progesterone deprivation impairs alveolar formation and fluid clearance in newborn piglets. AB - Exposure to high levels of estradiol (E2) and progesterone (P) derived from the fetoplacentomaternal unit during the last trimester of pregnancy may play a crucial role in prenatal lung development and immediate postnatal alveolar fluid clearance (AFC). To measure prenatal alveolar formation and postnatal amiloride sensitive AFC after pharmacological deprivation of E2 and P in utero, fetuses from five sows received an intramuscular depot injection of the E2 receptor blocker ICI 182.780 (ICI) and the P receptor blocker RTI 3021-022 (RTI) and fetuses of five other sows received a placebo injection (control group) during a laparotomy at 90 d of gestation (term gestation, 115 d). Piglets were delivered by cesarean section on d 114 of gestation. Of 95 live-born piglets, 35 were mechanically ventilated. The airways of the right lower lobe were isolated by a balloon catheter wedged in the bronchus and 5% albumin in 0.9% NaCl with or without 1 mmol/L amiloride was instilled. Amiloride-sensitive AFC was calculated from the protein concentration changes in fluid recovered after 120 min as the percentage of absorbed fluid. Lungs were removed under standardized conditions to perform alveolar counts. Prenatal treatment with ICI and RTI resulted in a significantly lower amiloride-sensitive AFC (median, 31%; min-max, -4-58) than placebo (74%, 18-231). Median alveolar counts per visual field were significantly lower in piglets that were exposed to ICI and RTI (38, 21-78) compared with placebo (56, 32-113). We conclude that prenatal E2 and P deprivation significantly impaired alveolar formation and amiloride-sensitive AFC. PMID- 16690947 TI - New evidence of effects of organophosphate pesticides on neurodevelopment in children: commentary on the article by Kofman et al. on page 88. PMID- 16690948 TI - Purified neonatal plasmacytoid dendritic cells overcome intrinsic maturation defect with TLR agonist stimulation. AB - Neonates are more susceptible than adults to viral and bacterial diseases. We hypothesized that plasmacytoid dendritic cells, the cells that provide large amounts of IFN-alpha in response to Toll-like receptor 9 (TLR9) agonists, are defective in neonates. To assess the intrinsic functionality of plasmacytoid dendritic cells from neonates we compared IFN-alpha production by plasmacytoid dendritic cells derived from neonates versus adults in both whole blood and in purified plasmacytoid dendritic cells. TLR9-stimulation of whole blood from adults and neonates resulted in comparable amounts of IFN-alpha production. However, we observed small but significant differences in IFN-alpha production from purified CD123+ plasmacytoid dendritic cells from neonates after stimulation with the TLR9 ligand CpG-DNA. Furthermore, we assessed surface expression of co stimulatory molecules on plasmacytoid dendritic cells after stimulation. While purified CD123+ plasmacytoid dendritic cells from adults up-regulated co stimulatory molecules CD80 and CD86 with IL-3 alone those from neonates required the addition of CpG-DNA to reach adult levels. Therefore, the intrinsic deficiencies of neonatal plasmacytoid dendritic cells can be mitigated by TLR9 agonists. These results are consistent with the observation that vaccines that effect strong adjuvant activity on dendritic cells can induce protective responses in neonates. PMID- 16690949 TI - Cognition- and anxiety-related behavior, synaptophysin and MAP2 immunoreactivity in the adult rat treated with a single course of antenatal betamethasone. AB - We investigated the effects of a single course of antenatal betamethasone on cognition- and anxiety-related behavior and synaptophysin and microtubule associated protein 2 (MAP2) immunoreactivity in the adult rat hippocampus. On d 20 of gestation, pregnant rats were injected with either 1) 170 microg/kg body weight of betamethasone ("clinically equivalent dose," equivalent to 12 mg twice, 24 h apart); 2) half this dose; or 3) vehicle. Cognition- and anxiety-related behavior of the offspring was analyzed at an age of 5 mo using the Morris water maze, object recognition task, and open field test. Subsequently, synaptophysin and MAP2 immunoreactivity were measured in the hippocampus. We report no detrimental effects of antenatal betamethasone on cognition- and anxiety-related behavior and synaptophysin immunoreactivity in the adult rat. On the other hand, MAP2 immunoreactivity was decreased by betamethasone in males, suggesting a permanent impairment in the hippocampus. Interestingly, the lower dose appears to have less influence in terms of growth restriction, known to be associated with an increased risk of disease in adulthood. Further research might elucidate whether the betamethasone effect on hippocampal neurons persists later in life and could affect the aging process increasing the risk for neuropathology of the adult. PMID- 16690950 TI - Unconjugated bilirubin modulates the intestinal epithelial barrier function in a human-derived in vitro model. AB - Unconjugated bilirubin promotes intestinal secretion without affecting nutrient digestion or absorption. In the current study, the effects of unconjugated bilirubin (UCB) on the barrier function of the intestinal epithelium were investigated. The apical side of human intestinal cell line Caco-2 monolayers was challenged with purified UCB. Transepithelial electrical resistance and paracellular fluxes of 10 kD Cascade blue conjugate dextran were measured. Cell monolayer viability was studied using LDH release and trypan blue exclusion tests. Redistribution of enterocyte tight junction occludin was studied by confocal microscopy. Bilirubin induced a dose-dependent decrease of transepithelial electrical resistance (TEER). This effect was maximal at 6 h and tended to be reversed at 48 h. Oxidated bilirubin was ineffective. Bilirubin significantly increased fluorescent dextran paracellular passage. Cell viability was not affected by UCB over the 5-200 nmol/L concentration range. Finally, bilirubin triggered a reversible redistribution of tight junctional occludin. UCB increases the permeability of intestinal epithelium. This effect is reversible, dependent on the redox status of the molecule and the rearrangement of the tight junction. These data attribute to bilirubin a novel role of functional modulator of intestinal paracellular permeability in vitro. PMID- 16690951 TI - Human congenital infection with Trypanosoma cruzi induces phenotypic and functional modifications of cord blood NK cells. AB - We studied the phenotype and activity of cord blood natural killer (NK) cells in newborns congenitally infected with Trypanosoma cruzi. We found that the proportion of CD56(bright) NK cells was significantly decreased in cord blood from these newborns, suggesting they may have been recruited to secondary lymphoid organs. The remaining CD56(bright) NK cells exhibited a defective ability in the production of interferon (IFN)-gamma following in vitro activation with interleukin (IL)-12 + IL-2 or IL-12 + IL-15 cytokines, as compared with NK cells from uninfected newborns. In addition, cord blood NK cells from congenitally infected newborns stimulated with cytokines have a decreased release of granzyme B (GrB) when incubated with K562 target cells. This defect in cytotoxic effector function is associated with a reduced surface expression of activating NK receptors (NKp30, NKp46, and NKG2D) on CD56(dim) NK cells compared with uninfected newborns. These alterations of fetal NK cells from congenitally infected newborns may reflect a down-regulation of the NK cell response after an initial peak of activation and could also be the result of T. cruzi modulating the immune response. PMID- 16690952 TI - Reduction in cerebellar volumes in preterm infants: relationship to white matter injury and neurodevelopment at two years of age. AB - A substantial number of prematurely born infants will experience later neurodevelopmental challenges. Abnormal development of the cerebellum may be related to some of the impairments exhibited by preterm children. To test the hypothesis that cerebellar development is structurally impaired in preterm infants and associated with adverse outcomes, we studied 83 preterm infants and 13 term controls using volumetric magnetic resonance imaging techniques to obtain cerebellar volumes (CV) at term corrected and subsequent neurodevelopmental assessment at 2 y of age. The preterm group had smaller mean CV at term compared with the term control infants [mean (SD) CV, 22.0 (5.0) versus 23.5 (5.0) cc; mean difference (95% confidence interval), 1.5 (-1.5, 4.4)] although this did not reach statistical significance. Within the preterm group, there was evidence of a reduction in CV related to the presence of white matter injury (WMI) after adjusting for intracranial volume (ICV) [WMI grade 1 versus grade 2: mean (SD) CV, 23.6 (5.0) versus 21.6 (4.5); p = 0.01; WMI grade 1 versus grade 3 and 4: 23.6 (5.0) versus 20.8 (5.6); p = 0.07]. Within the preterm infants, there was no apparent relationship between CV at term and gestational age at birth after adjusting for ICV. At 2 y of age, CV showed a weak correlation with cognitive and motor development, although this was principally mediated by WMI. In conclusion, we found no evidence for a primary impairment in cerebellar development in relation to prematurity, although there was evidence for a secondary effect of cerebral WMI on cerebellar development independent of immaturity. PMID- 16690953 TI - Saccharomyces boulardii produces in rat small intestine a novel protein phosphatase that inhibits Escherichia coli endotoxin by dephosphorylation. AB - Using a polyclonal antibody raised against a highly conserved sequence of 38 amino acids containing the activation site (VTDSAAGAT) common to mammalian and yeast alkaline phosphatases (AP), we identified in decapsidated Saccharomyces boulardii a protein phosphatase detected by autoradiography as a single signal (63 kD). Using an affinity chromatography column, the protein phosphatase could be concentrated 39.1-fold and presented as a doublet of two subunits. Compared with rat and bovine purified intestinal AP, the enzyme from S. boulardii had a greater ability to dephosphorylate the lipopolysaccharide (LPS) of Escherichia coli 055B5. When tested in vivo, intraperitoneal injection of intact LPS to rats produced, after 9 h, 100 ng/mL of circulating tumor necrosis factor-alpha with inflammatory lesions and apoptotic bodies in the liver and the heart, whereas rats injected with partially dephosphorylated LPS produced only 40 ng/mL tumor necrosis factor-alpha without organic lesions. In conclusion, S. boulardii is able to inhibit toxicity of E. coli surface endotoxins by the release of a protein phosphatase exhibiting a great capacity of dephosphorylation. PMID- 16690954 TI - Moving toward the light: using new technology to answer old questions. AB - Fluorescence microscopy has become a principle methodology in the field of developmental biology. Recent technological advances have led to the design of high-speed and high-resolution confocal and multiphoton microscopes that enable researchers to obtain three- and four-dimensional information in living cells and whole embryos. Paralleling this progress, the development of stable and bright vital fluorescent probes has revolutionized the ability to track individual cells in vitro and in vivo and to visualize intercellular and subcellular molecular interactions in real time. Combining imaging modalities and labeling techniques that are increasingly unobtrusive to cell and whole animal function, our understanding of how proteins interact, tissues take form, and organs synchronize to create a functioning animal is reaching a whole new level. PMID- 16690955 TI - Quantifying cardiovascular flow dynamics during early development. AB - The relationship between developing biologic tissues and their dynamic fluid environments is intimate and complex. Increasing evidence supports the notion that these embryonic flow-structure interactions influence whether development will proceed normally or become pathogenic. Genetic, pharmacological, or surgical manipulations that alter the flow environment can thus profoundly influence morphologic and functional cardiovascular phenotypes. Functionally deficient phenotypes are particularly poorly described as there are few imaging tools with sufficient spatial and temporal resolution to quantify most intra-vital flows. The ability to visualize biofluids flow in vivo would be of great utility in functionally phenotyping model animal systems and for the elucidation of the mechanisms that underlie flow-related mechano-sensation and transduction in living organisms. This review summarizes the major methodological advances that have evolved for the quantitative characterization of intra-vital fluid dynamics with an emphasis on assessing cardiovascular flows in vertebrate model organisms. PMID- 16690956 TI - Porcine pulmonary artery and bronchial responses to endothelin-1 and norepinephrine on recovery from hypoxic pulmonary hypertension. AB - Many infants recovering from acute lung disease and pulmonary hypertension still have evidence of reactive airways disease at one year of age, suggesting longer term airway effects. We hypothesized that parallel changes in smooth muscle would occur in airways and pulmonary arteries from animals with pulmonary hypertension and during normoxic recovery. Thus, two-hour-old piglets were subjected to 3 d chronic hypobaric hypoxia and 3-d-old piglets were subjected to 11 d hypoxia. Some animals were allowed to recover in room air for 3 or 6 d. The amount of smooth muscle and responses of isolated paired bronchial and pulmonary artery rings to endothelin-1 (ET-1) and norepinephrine were studied at the end of hypoxic exposure, on recovery and in age-matched control animals. In all hypoxia induced pulmonary hypertensive animals, smooth muscle area and ET-1 contractile response was increased in the pulmonary arteries and bronchi. Norepinephrine induced relaxant response was impaired significantly in both bronchi and pulmonary arteries. After 3 d recovery, pulmonary arterial smooth muscle area decreased by 65%, and ET-1-induced contractile responses were normal for age. In the airways, ET-1 contractile response only normalized after six days and bronchial smooth muscle was still increased. After 6 d recovery pulmonary arterial norepinephrine-induced relaxant response had returned to normal, but bronchial response remained impaired. Thus during pulmonary hypertension, both bronchial and pulmonary arterial smooth muscle area and contractile responses are increased. On recovery, regression of bronchial structural and functional abnormalities is slower than in pulmonary arteries. PMID- 16690957 TI - Calculated in vivo free bilirubin levels in the central nervous system of Gunn rat pups. AB - In vitro studies suggest a free bilirubin (B(F)) concentration in the range of 71 770 nmol/L can induce neurotoxicity. In vivo data regarding central nervous system (CNS) B(F) levels have not been determined. We calculated in vivo CNS B(F) levels in Gunn rat pups (15-19 d old; heterozygous nonjaundiced Gunn rats (J/j) and homozygous jaundiced Gunn rats (j/j); saline or sulfadimethoxine treated) based on 1) total brain bilirubin (TBB) content, 2) brain albumin level, 3) CNS bilirubin binding capacity attributable to brain albumin determined using an ultrafiltration technique, and 4) published Gunn rat albumin-bilirubin binding constants (k). Gunn rat brain bilirubin binding capacity was approximately 22 x 10(-3) micromol/g, of which two thirds was accounted for by brain albumin. Using a Gunn rat pup in vivo, k of 9.2 L/micromol, calculated CNS B(F) levels ranged from 72 to 112 nmol/L [95% confidence interval (CI)] in saline and from 59 to 156 nmol/L (95% CI) in sulfadimethoxine-treated J/j pups. These animals demonstrated no neurobehavioral abnormalities and normal cerebellar weight. Calculated CNS B(F) levels were severalfold higher (p < 0.001) in saline (95% CI: 556-1110 nmol/L) and sulfadimethoxine-treated (95% CI: 3461-8985 nmol/L) j/j pups; the former evidenced reduced cerebellar weight; the latter both reduced cerebellar weight and acute neurobehavioral abnormalities. We conclude that calculated CNS B(F) values in j/j pups are substantially higher than those in J/j animals. Given the absence of CNS abnormalities in J/j pups, the presence of such in j/j animals, and the CNS B(F) levels in these groups, we speculate that the CNS B(F) neurotoxicity threshold in vivo is subsumed within the range (71-770 nmol/L) reported in vitro. PMID- 16690958 TI - Monitoring of ketogenic diet for carnitine metabolites by subcutaneous microdialysis. AB - The ketogenic diet (KD) provides ketones from the degradation of free fatty acids for energy metabolism. It is a therapeutic option for pharmacoresistant epilepsies. Carnitine is the carrier molecule that transports fatty acids across the mitochondrial membrane for degradation into ketones. The integrity of this transport system is a prerequisite for an adequate ketogenic response. For monitoring of tissue metabolism with KD, we used the sampling method of s.c. microdialysis (MD), which permits minimally invasive, frequent, and extensive metabolic monitoring independent of blood tests. By using this new method, we monitored changes in carnitine metabolism induced by KD, particularly in free carnitine (C0), acetylcarnitine (C2), and hydroxybutyrylcarnitine (C4OH). Correlation of microdialysate and tissue concentrations for carnitines in vitro was about 85%. Carnitine metabolism was monitored in seven children started on a KD for pharmacoresistant epilepsy after a conventional initial fasting period. Detected metabolic changes consisted of a slight decrease in s.c. C0 and a marked increase in C2/CO and C4OH/CO levels. The levels of s.c. C4OH strongly correlate with beta-hydroxybutyrate (beta-OHB) levels in plasma providing an additional parameter for the carnitine reserve of the body and reflect an optimal ketogenic energy supply. Subcutaneous MD allows close and extensive monitoring of metabolism with a KD. PMID- 16690959 TI - Imaging tools for the developmental biologist: ultrasound biomicroscopy of mouse embryonic development. AB - Progress has been rapid in the elucidation of genes responsible for cardiac development. Strategies to ascertain phenotypes, however, have lagged behind advances in genomics, particularly in the in vivo mouse embryo, considered a model organism for mammalian development, and for human development and disease. Over the past several years, our laboratory and others have pioneered a variety of ultrasound biomicroscopy (UBM)-Doppler approaches to study in vivo development in both normal and mutant mouse embryos. This state-of-the-art review will discuss the development and potential of ultrasound biomicroscopy as a tool for the in vivo imaging and phenotyping of both cardiac and non-cardiac organ systems in the early developing mouse. Broad, long-term research objectives are to define living structure-function relationships during critical periods of mammalian morphogenesis. PMID- 16690960 TI - Role of chymase-dependent angiotensin II formation in monocrotaline-induced pulmonary hypertensive rats. AB - Angiotensin II-forming chymase is expressed in the pulmonary arteries of the monocrotaline-induced pulmonary hypertensive rats, but its actual role is unclear. We studied chymase-dependent angiotensin II formation in the pulmonary arteries of the monocrotaline-induced pulmonary hypertensive rats and observed the effects of an angiotensin II receptor blocker on vascular remodeling. Four weeks after the administration of monocrotaline (60 mg/kg, s.q.), echocardiographic, hemodynamic, morphometric and biochemical analyses were performed. Age-matched rats were used as controls. To evaluate the effects of an angiotensin II receptor blocker, 2 wk after beginning of monocrotaline treatment, the rats were given candesartan (10 mg/kg per day) or placebo for 2 wk. In the monocrotaline-induced pulmonary hypertensive rats, the elevated systolic pulmonary arterial pressure and right ventricular hypertrophy were observed. Medial hypertrophy of lung arterioles was also observed. Chymase activity and angiotensin II concentration, but not angiotensin-converting enzyme activity, were significantly increased in the lung. In the angiotensin II receptor blocker treated group, both systolic pulmonary arterial pressure and right ventricular hypertrophy were significantly reduced, and arteriolar hypertrophy was also prevented. Thus, angiotensin II-forming chymase may play a role in the proliferation of the medial layer in the lung arterioles of monocrotaline-induced pulmonary hypertensive rats. PMID- 16690961 TI - Perfluorochemical liquids enhance delivery of superoxide dismutase to the lungs of juvenile rabbits. AB - Previous studies suggest acute lung injury (ALI) in premature newborns is associated with relative deficiency of antioxidant enzymes that may be ameliorated by recombinant human superoxide dismutase (rhSOD). Perfluorochemicals (PFCs) are distributed homogeneously and support gas exchange in diseased lungs. We investigated whether PFCs could provide an effective delivery system for rhSOD. Juvenile rabbits were lung-lavaged, treated with surfactant, and randomized: group I: fluorescently labeled rhSOD (5 mg/kg in 2 mL/kg saline); group II: fluorescently labeled rhSOD (5 mg/kg in 18 mL/kg PFC). Animals were ventilated with oxygen for 4 h; the lungs were harvested for analysis of SOD distribution and oxidative injury. Cardiopulmonary indices remained stable and similar between groups. Qualitative assessment (QA) showed a more homogeneous lung SOD distribution in group II and a better histologic profile. QA of lung SOD distribution showed significant increase in SOD concentrations in group II (7.37 +/- 1.54 microg/mg protein) compared with group I (1.65 +/- 0.23 microg/mg protein). Oxidative injury as assessed by normalized protein carbonyl was 149.1 +/- 26.8% SEM in group II compared with 200.5 +/- 7.3% SEM in group I. Plasma SOD was significantly higher in group II. Administration of rhSOD with or without PFCs does not compromise cardiovascular function or impede lung recovery after ALI. PFCs enhance rhSOD delivery to the lungs by 400% while decreasing lung oxidative damage by 25% compared with rhSOD alone. These data suggest that PFCs optimize lung rhSOD delivery and might enhance the beneficial effects of rhSOD in preventing acute and chronic lung injury. PMID- 16690962 TI - The influence of gestation and mechanical ventilation on serum clara cell secretory protein (CC10) concentrations in ventilated and nonventilated newborn infants. AB - Clara cell secretory protein (CC10) is an important anti-inflammatory mediator in the adult lung, but its role in newborn pulmonary protection is uncertain. We examined the early postnatal behavior of CC10 in newborn serum and tracheal fluid and hypothesized that CC10 production is positively influenced by gestation. Blood from 165 infants from the first, third/fourth, and seventh days of life (gestational ages: 23-29 wk, 30-36 wk, >36 wk) and tracheal fluid (TF) from the first day of life from 32 ventilated infants were analyzed for CC10. Surfactant proteins A (SPA) and B (SPB) were also analyzed from the blood of a subgroup of infants. Serum CC10 on day 1 was highest in term infants (69.4 ng/mL), followed by moderately preterm (55.8 ng/mL), and then extremely preterm infants (median 42.1 ng/mL). Term infants also had higher tracheal fluid CC10 than preterm infants. (20.152 ng/mL versus 882 ng/mL). Mechanical ventilation increased serum CC10 only in moderately preterm infants, and only on d 1 [68.4 ng/mL versus 42.1 ng/mL (nonventilated moderately preterm infants)]. Serum CC10 decreased progressively by the end of the first week in all infants, in contrast to SPA and SPB, which increased. Our results show that CC10 is detectable in the blood of newborn infants and that a production surge occurs at birth. This surge is more pronounced in term infants and may confer them with superior extrauterine pulmonary protection compared with preterm infants. PMID- 16690963 TI - Mantle cell lymphoma cells express predominantly cyclin D1a isoform and are highly sensitive to selective inhibition of CDK4 kinase activity. AB - The prognosis for patients with mantle cell lymphoma (MCL) is poor, and at present there is no truly effective therapy. Gene translocation-mediated constitutive expression of cyclin D1 seems to play the key role in the pathogenesis of MCL. Here we report that although 3 of 4 MCL cell lines expressed the recently identified, highly oncogenic cyclin D1b isoform, as well as the canonical cyclin D1a, 8 MCL patient samples expressed only the cyclin D1a protein despite expressing detectable cyclin D1b mRNA. Cell lines and tissue samples displayed constitutive activation of the cyclin D1 signaling cascade, as evidenced by strong expression of CDK4, Rb phosphorylation, and cyclin D1/CDK4 coassociation. All MCL cell lines and tissues examined displayed nondetectable to diminished expression of the cyclin D1 inhibitor p16. Novel small molecule CDK4/CDK6 inhibitor PD0332991 profoundly suppressed--at low nanomolar concentrations--Rb phosphorylation, proliferation, and cell cycle progression at the G0/G1 phase of MCL cells. These findings provide evidence that MCL should be very sensitive to targeted therapy aimed at functional inhibition of the cyclin D1/CDK4 complex. PMID- 16690964 TI - Platelet-derived growth factor receptor-beta promotes early endothelial cell differentiation. AB - Platelet-derived growth factor BB (PDGF-BB) has been assigned a critical role in vascular stability by promoting the recruitment of PDGF receptor-beta-expressing perivascular cells. Here we present data indicating that early hematopoietic/endothelial (hemangio) precursors express PDGFR-beta based on coexpression with CD31, vascular endothelial growth factor receptor-2, and CD41 in 2 models: mouse yolk sac (embryonic day 8 [E8]) and differentiating mouse embryonic stem cells (embryoid bodies). Expression of PDGFR-beta on hemangioprecursor cells in the embryoid bodies gradually disappeared, and, at E14, expression appeared on perivascular cells. Activation of the PDGFR-beta on the hemangioprecursors accelerated the differentiation of endothelial cells, whereas differentiation of the hematopoietic lineage was suppressed. In E9.5 yolk sacs derived from recombinant mice expressing kinase-active PDGFR-beta with an aspartic acid to asparagine (D894N) replacement in the kinase activating loop and from mice with ubiquitous expression of PDGF-BB driven by the Rosa26 locus, the number of CD41-expressing early hematopoietic cells decreased by 36% and 34%, respectively, compared with staged wild-type littermates. Moreover, enhanced vascular remodeling was evident in the Rosa26-PDGF-BB yolk sacs. We conclude that PDGFR-beta is expressed on early hemangioprecursor cells, regulating vascular/hematopoietic development. PMID- 16690965 TI - Apoptotic cells promote macrophage survival by releasing the antiapoptotic mediator sphingosine-1-phosphate. AB - Programmed cell death is vital for a number of pathophysiologic settings. Apoptotic cells are rapidly engulfed by phagocytes (ie, macrophages), which in turn acquire an anti-inflammatory phenotype known as alternative activation or the M2-type. Here we show that interaction of apoptotic cells with macrophages attenuates cell death pathways in the latter. Protection of human macrophages required phosphoinositide 3-kinase (PI3K), extracellular signal-regulated kinase 1/2 (ERK1/2), and Ca2+ signaling, and correlated with Bcl-X(L) and Bcl-2 up regulation as well as Ser136-Bad phosphorylation. Unexpectedly, neither phagocytosis nor binding of apoptotic debris to the phagocyte was necessary to induce protection. Surprisingly, apoptotic cells released sphingosine-1-phosphate (S1P), mainly derived from sphingosine kinase 2, as a survival messenger. This points to an active role of apoptotic cells in preventing cell destruction in their neighborhood, with implications for innate immunity and inflammation. PMID- 16690966 TI - Targeted gene deletion demonstrates that the cell adhesion molecule ICAM-4 is critical for erythroblastic island formation. AB - Erythroid progenitors differentiate in erythroblastic islands, bone marrow niches composed of erythroblasts surrounding a central macrophage. Evidence suggests that within islands adhesive interactions regulate erythropoiesis and apoptosis. We are exploring whether erythroid intercellular adhesion molecule 4 (ICAM-4), an immunoglobulin superfamily member, participates in island formation. Earlier, we identified alpha(V) integrins as ICAM-4 counterreceptors. Because macrophages express alpha(V), ICAM-4 potentially mediates island attachments. To test this, we generated ICAM-4 knock-out mice and developed quantitative, live cell techniques for harvesting intact islands and for re-forming islands in vitro. We observed a 47% decrease in islands reconstituted from ICAM-4 null marrow compared to wild-type marrow. We also found a striking decrease in islands formed in vivo in knock-out mice. Further, peptides that block ICAM-4/alpha(V) adhesion produced a 53% to 57% decrease in reconstituted islands, strongly suggesting that ICAM-4 binding to macrophage alpha(V) functions in island integrity. Importantly, we documented that alpha(V) integrin is expressed in macrophages isolated from erythroblastic islands. Collectively, these data provide convincing evidence that ICAM-4 is critical in erythroblastic island formation via ICAM-4/alpha(V) adhesion and also demonstrate that the novel experimental strategies we developed will be valuable in exploring molecular mechanisms of erythroblastic island formation and their functional role in regulating erythropoiesis. PMID- 16690967 TI - Predictive factors for thrombosis and major bleeding in an observational study in 181 patients with heparin-induced thrombocytopenia treated with lepirudin. AB - The antithrombotic efficacy of lepirudin in patients with heparin-induced thrombocytopenia (HIT) is compromised by an increased risk for bleeding. A retrospective observational analysis in 181 patients (median age, 67 years) with confirmed HIT treated in routine practice with lepirudin was performed to identify predictive factors for thrombotic and bleeding complications. Lepirudin was administered at a mean (+/- SD) dose of 0.06 +/- 0.04 mg/kg/h (compared with a recommended initial dose of 0.15 mg/kg/h). Mean activated partial thromboplastin time was greater than 1.5 times baseline value in 99.4% of patients. Median treatment duration was 7.7 days. Until discharge from the hospital, 13.8% and 20.4% of patients experienced a thrombotic or a major bleeding event, respectively. On multivariate analysis, mean lepirudin dose was not a significant predictive factor for thrombosis. In contrast, mean lepirudin dose greater than 0.07 mg/kg/h, long duration of lepirudin treatment, and moderate to severe renal impairment were significant positive factors for major bleeding. Overall, these results suggest that the recommended dose of lepirudin in patients with HIT is too high; the use of reduced doses may be safer with regard to bleeding risk and does not compromise antithrombotic efficacy. PMID- 16690968 TI - The Follicular Lymphoma International Prognostic Index (FLIPI) separates high risk from intermediate- or low-risk patients with advanced-stage follicular lymphoma treated front-line with rituximab and the combination of cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) with respect to treatment outcome. AB - The Follicular Lymphoma International Prognostic Index (FLIPI) was developed to predict prognosis of patients with follicular lymphoma (FL). However, it was based on different protocols, none of which included rituximab. The current analysis aimed at evaluating the predictive value of the FLIPI for treatment outcome in 362 patients with advanced-stage FL treated front-line with rituximab/CHOP in a prospective trial of the German Low Grade Lymphoma Study Group. According to the FLIPI, 14% of the patients were classified as low-risk, 41% as intermediate-risk, and 45% as high-risk patients. With a 2-year time to treatment failure (TTF) of 67%, high-risk patients had a significantly shorter TTF as compared with low- or intermediate-risk patients (2-year TTF of 92% and 90%, respectively; P < .001). Our data demonstrate that the FLIPI is able to identify high-risk patients with advanced-stage FL after first-line treatment with rituximab/chemotherapy. PMID- 16690969 TI - Asthma is associated with acute chest syndrome and pain in children with sickle cell anemia. AB - Pain and acute chest syndrome (ACS) episodes are 2 of the most common causes of hospitalization in children with sickle cell anemia (SCA). However, very few potentially modifiable risk factors for either condition have been identified. In this prospective infant cohort study, we tested the hypothesis that asthma is associated with an increased incidence rate of pain and ACS episodes. An infant cohort was composed of 291 African American children with hemoglobin SS enrolled in the Cooperative Study for Sickle Cell Disease before age 6 months and followed beyond age 5 years. Asthma was defined by a physician diagnosis, an acute asthma event, or use of prescription asthma medications. The incidence rates of ACS and painful episodes were compared for children with and without asthma. A clinical diagnosis of asthma was made in 17% of the cohort. Asthma was associated with more frequent ACS episodes (0.39 vs 0.20 events per patient year, P < .001) and painful episodes (1.39 vs 0.47 events per patient year, P < .001). In conclusion, in children with SCA, asthma is associated with an increased incidence of sickle cell disease-related morbidity, including ACS and painful episodes. PMID- 16690970 TI - Donor-derived mesenchymal stem cells are immunogenic in an allogeneic host and stimulate donor graft rejection in a nonmyeloablative setting. AB - Mesenchymal stem cells (MSCs) are multipotent progenitor cells that have emerged as a promising tool for clinical application. Further clinical interest has been raised by the observation that MSCs are immunoprivileged and, more important, display immunosuppressive capacities. These properties may be of therapeutic value in allogeneic transplantation to prevent graft rejection and to prevent and treat graft-versus-host disease. In the present study, we examined the in vivo immunomodulatory properties of MSCs in murine models of allogeneic bone marrow (BM) transplantation. Sublethally irradiated recipients received allogeneic BM with or without host or donor MSCs. The addition of host MSCs significantly enhanced the long-term engraftment associated with tolerance to host and donor antigens. However, the infusion of donor MSCs was associated with significantly increased rejection of allogeneic donor BM cells. Moreover, we showed that the injection of merely allogeneic donor MSCs in naive mice was sufficient to induce a memory T-cell response. Although the observed engraftment-promoting effects of host MSCs in vivo support the therapeutic potential of MSCs, our results also indicate that allogeneic MSCs are not intrinsically immunoprivileged and that under appropriate conditions, allogeneic MSCs induce a memory T-cell response resulting in rejection of an allogeneic stem cell graft. PMID- 16690971 TI - Interstitial uniparental isodisomy at clustered breakpoint intervals is a frequent mechanism of NF1 inactivation in myeloid malignancies. AB - To identify the mechanism of loss of heterozygosity (LOH) and potential modifier gene(s), we investigated the molecular basis of somatic NF1 inactivation in myeloid malignancies from 10 children with neurofibromatosis type 1. Loci across a minimal 50-Mb region of primarily the long arm of chromosome 17 showed LOH in 8 cases, whereas a less than 9-Mb region of loci flanking NF1 had LOH in the remaining 2 cases. Two complementary techniques, quantitative polymerase chain reaction (PCR) and fluorescence in situ hybridization (FISH), were used to determine whether the copy number at loci that showed LOH was 1 or 2 (ie, deleted or isodisomic). The 2 cases with LOH limited to less than 9 Mb were intrachromosomal deletions. Among the 8 leukemias with 50-Mb LOH segments, 4 had partial uniparental isodisomy and 4 had interstitial uniparental isodisomy. These isodisomic cases showed clustering of the centromeric and telomeric LOH breakpoints. This suggests that the cases with interstitial uniparental isodisomy arose in a leukemia-initiating cell by double-homologous recombination events at intervals of preferred mitotic recombination. Homozygous inactivation of NF1 favored outgrowth of the leukemia-initiating cell. Our studies demonstrate that LOH analyses of loci distributed along the chromosomal length along with copy number analysis can reveal novel mechanisms of LOH that may potentially identify regions harboring "cryptic" tumor suppressor or modifier genes whose inactivation contributes to tumorigenesis. PMID- 16690972 TI - Improved targeting of miRNA with antisense oligonucleotides. AB - MicroRNAs (miRNAs) are a class of 20-24 nt noncoding RNAs that regulate target mRNAs post-transcriptionally by binding with imperfect complementarity in the 3' untranslated region (3'-UTR) and inhibiting translation or RNA stability. Current understanding of miRNA biology is limited, and antisense oligonucleotide (ASO) inhibition is a powerful technique for miRNA functionalization in vitro and in vivo, and for therapeutic targeting of miRNAs. Identification of optimal ASO chemistries for targeting miRNAs is therefore of great interest. We evaluated a number of 2'-sugar and backbone ASO modifications for their ability to inhibit miR-21 activity on a luciferase reporter mRNA. ASO modifications that improved target affinity improved miRNA ASO activity, yet the positioning of high-affinity modifications also had dramatically different effects on miRNA activity, suggesting that more than affinity determined the effectiveness of the miRNA ASOs. We present data in which the activity of a modified miRNA ASO was inversely correlated to its tolerability as an siRNA passenger strand, suggesting that a similar mechanism could be involved in the dissociation of miRNA ASOs and siRNA passenger strands. These studies begin to define the factors important for designing improved miRNA ASOs, enabling more effective miRNA functionalization and therapeutic targeting. PMID- 16690973 TI - Regional pulmonary perfusion, inflation, and ventilation defects in bronchoconstricted patients with asthma. AB - RATIONALE: Bronchoconstriction in asthma leads to heterogeneous ventilation and the formation of large and contiguous ventilation defects in the lungs. However, the regional adaptations of pulmonary perfusion (Q) to such ventilation defects have not been well studied. METHODS: We used positron emission tomography to assess the intrapulmonary kinetics of intravenously infused tracer nitrogen-13 ((13)NN), and measured the regional distributions of ventilation and perfusion in 11 patients with mild asthma. For each subject, the regional washout kinetics of (13)NN before and during methacholine-induced bronchoconstriction were analyzed. Two regions of interest (ROIs) were defined: one over a spatially contiguous area of high tracer retention (TR) during bronchoconstriction and a second one covering an area of similar size, showing minimal tracer retention (NR). RESULTS: Both ROIs demonstrated heterogeneous washout kinetics, which could be described by a two-compartment model with fast and slow washout rates. We found a systematic reduction in regional Q to the TR ROI during bronchoconstriction and a variable and nonsignificant change in relative Q for NR regions. The reduction in regional Q was associated with an increase in regional gas content of the TR ROI, but its magnitude was greater than that anticipated solely by the change in regional lung inflation. CONCLUSION: During methacholine-induced bronchoconstriction, perfusion to ventilation defects are systematically reduced by a relative increase in regional pulmonary vascular resistance. PMID- 16690974 TI - Pulmonary vascular endothelial growth factor-C in development and lung injury in preterm infants. AB - RATIONALE: In mice, vascular endothelial growth factor-C (VEGF-C) plays an important role in development of the lymphatic system and in pathogenesis of pulmonary inflammation. Its role in development of the lymphatic system in human lung and in lung injury in newborns remains unclear. OBJECTIVES: We studied the role of VEGF-C in developing human lung, and in acute and chronic lung injury in preterm infants. METHODS: Included in the immunohistochemistry study were 10 fetuses, 15 control neonates without primary lung disease, 15 preterm infants with respiratory distress syndrome, and 8 infants with bronchopulmonary dysplasia. Tracheal aspirate fluid samples of intubated very-low-birth-weight infants during Postnatal Weeks 1-5 were analyzed with ELISA. RESULTS: Bronchiolar staining for VEGF-C was observed in all 48 samples. Alveolar epithelial staining was seen in most fetuses (8/10). In addition, staining was observed in alveolar macrophages in bronchopulmonary dysplasia (4/8), and late respiratory distress syndrome (2/7). VEGF receptor-3 (VEGFR-3) staining was observed in lymphatic endothelium adjacent to vascular endothelium. VEGF-C was expressed consistently in tracheal aspirate fluid, being highest during the first 2 postnatal days. Antenatal administration of glucocorticoids was associated with higher VEGF-C in tracheal aspirate fluid. CONCLUSIONS: The pattern of pulmonary VEGF-C and VEGFR-3 protein expression and consistent VEGF-C protein appearance in tracheal aspirate fluid in human preterm infants indicate a role for VEGF-C in the physiologic development of the lymphatic system of the lung. PMID- 16690975 TI - Variants in the glutamate-cysteine-ligase gene are associated with cystic fibrosis lung disease. AB - BACKGROUND: Chronic progressive lung disease is the most serious complication of cystic fibrosis (CF). Glutathione plays an important role in the protection of the CF lung against oxidant-induced lung injury. OBJECTIVES: We hypothesized that a polymorphism in a novel candidate gene that regulates glutathione synthesis might influence CF lung disease. METHODS: In a cross-sectional study, subjects were recruited from CF clinics in Seattle and multiple centers in Canada. We tested for an association between CF lung disease and a functional polymorphism in the glutamate-cysteine ligase catalytic subunit (GCLC) gene. Multiple linear regression was used to test for association between polymorphisms of GCLC and severity of CF lung disease while adjusting for age, Pseudomonas aeruginosa infection, and cystic fibrosis transmembrane conductance regulator (CFTR) genotype. Analysis was repeated for patients with CF stratified by CFTR genotype. MEASUREMENTS AND MAIN RESULTS: A total of 440 subjects with CF participated in the study (51% male; mean [+/- SD] age, 26 +/- 11 yr; mean FEV(1), 62 +/- 28% predicted). In the total population, there was a trend toward an association between GCLC genotypes and CF lung disease (linear regression coefficient [SEM], 1.68 [1.0]; p = 0.097). In the stratified analysis, there was a highly significant association between GCLC genotype and CF lung function in subjects with a milder CFTR genotype (linear regression coefficient [SEM], 5.5 (1.7); p = 0.001). CONCLUSIONS: In patients with CF with a milder CFTR genotype, there is a strong association between functional polymorphisms of the GCLC gene and CF lung disease severity. PMID- 16690976 TI - Impaired performance in commercial drivers: role of sleep apnea and short sleep duration. AB - Sleepiness plays an important role in major crashes of commercial vehicles. Because determinants are likely to include inadequate sleep and sleep apnea, we evaluated the role of short sleep durations over 1 wk at home and sleep apnea in subjective sleepiness (Epworth Sleepiness Scale), objective sleepiness (reduced sleep latency as determined by the Multiple Sleep Latency Test), and neurobehavioral functioning (lapses in performance, tracking error in Divided Attention Driving Task) in commercial drivers. Studies were conducted in 247 of 551 drivers at higher risk for apnea and in 159 of 778 drivers at lower risk. A multivariate linear association between the sets of outcomes and risk factors was confirmed (p < 0.0001). Increases in subjective sleepiness were associated with shorter sleep durations but not with increases in severity of apnea. Increases in objective sleepiness and performance lapses, as well as poorer lane tracking, were associated with shorter sleep durations. Associations with sleep apnea severity were not as robust and not strictly monotonic. A significant linear association with sleep apnea was demonstrated only for reduced sleep latency. The effects of severe apnea (apnea-hypopnea index, at least 30 episodes/h), which occurred in 4.7%, and of sleep duration less than 5 h/night, which occurred in 13.5%, were similar in terms of their impact on objective sleepiness. Thus, addressing impairment in commercial drivers requires addressing both insufficient sleep and sleep apnea, the former being more common. PMID- 16690977 TI - Serial testing of health care workers for tuberculosis using interferon-gamma assay. AB - RATIONALE: Although interferon-gamma (IFN-gamma) assays are promising alternatives to the tuberculin skin test (TST), their serial testing performance is unknown. OBJECTIVE: To compare TST and IFN-gamma conversions and reversions in healthcare workers. METHODS: We prospectively followed-up 216 medical and nursing students in India who underwent baseline and repeat testing (after 18 mo) with TST and QuantiFERON-TB Gold In-Tube (QFT). TST conversions were defined as reactions greater than or equal to 10 mm, with increments of 6 or 10 mm over baseline. QFT conversions were defined as baseline IFN-gamma less than 0.35 and follow-up IFN-gamma greater than or equal to 0.35 or 0.70 IU/ml. QFT reversions were defined as baseline IFN-gamma greater than or equal to 0.35 and follow-up IFN-gamma less than 0.35 IU/ml. RESULTS: Of the 216 participants, 48 (22%) were TST-positive, and 38 (18%) were QFT-positive at baseline. Among 147 participants with concordant baseline negative results, TST conversions occurred in 14 (9.5%; 95% confidence interval [CI] = 5.3-15.5) using the 6 mm increment definition, and 6 (4.1%; 95% CI = 1.5-8.7) using the 10 mm increment definition. QFT conversions occurred in 17/147 participants (11.6%; 95% CI = 6.9-17.9) using the definition of IFN-gamma greater than or equal to 0.35 IU/ml, and 11/147 participants (7.5%; 95% CI = 3.8-13.0) using IFN-gamma greater than or equal to 0.70 IU/ml. Agreement between TST (10 mm increment) and QFT conversions (>or= 0.70 IU/ml) was 96% (kappa = 0.70). QFT reversions occurred in 2/28 participants (7%) with baseline concordant positive results, as compared with 7/10 participants (70%) with baseline discordant results (p < 0.001). CONCLUSIONS: IFN-gamma assay shows promise for serial testing, but repeat results need to be interpreted carefully. To meaningfully interpret serial results, the optimal thresholds to distinguish new infections from nonspecific variations must be determined. PMID- 16690978 TI - Discoidin domain receptor 1-deficient mice are resistant to bleomycin-induced lung fibrosis. AB - RATIONALE: Discoidin domain receptor 1 (DDR1) is a tyrosine kinase activated by native collagens. Based on previous findings showing increased DDR1 expression in bronchoalveolar lavage cells from patients with idiopathic pulmonary fibrosis, we hypothesized that DDR1 mediates disease progression after lung injury. OBJECTIVES: To investigate the inflammatory and fibrotic responses of DDR1 knockout and wild-type mice to bleomycin-induced lung injury. METHODS: Age- and sex-matched DDR1 knockout and wild-type C57BL/6 mice received a single intratracheal instillation of 2 U/kg bleomycin or saline, respectively. After 2 wk, lung inflammation and fibrosis were assessed using immunohistochemistry, real time polymerase chain reaction, TUNEL assay, ELISA, fluorescence-activated cell sorting, and Western blot analysis. MEASUREMENTS AND MAIN RESULTS: Compared with wild-type animals, DDR1-null mice were largely protected against bleomycin induced injury. Bleomycin-induced increases in collagen protein levels and tenascin-C mRNA levels were abrogated in knockout animals. Furthermore, myofibroblast expansion and apoptosis were much lower in these animals compared with their wild-type counterparts. Absence of inflammation in knockout mice was confirmed by lavage cell count and a cytokine ELISA. Western blot analysis of injured lung tissue revealed that DDR1-null mice failed to respond to the bleomycin insult with p38 MAPK activation, which was readily observed in wild type mice. CONCLUSIONS: DDR1 expression is a prerequisite for the development of lung inflammation and fibrosis. Blockade of DDR1 may therefore be a novel therapeutic intervention in patients with pulmonary fibrosis. PMID- 16690979 TI - Myocardial dysfunction and potential cardiac hypoxia in rats induced by carbon monoxide inhalation. AB - BACKGROUND: Results from both animal and human being studies provide evidence that inhalation of concentrations of carbon monoxide (CO) at around 100 ppm has antiinflammatory effects. These low levels of CO are incriminated in ischemic heart diseases experienced by cigarette smokers and, in some cases, from air pollution. Although neurologic mechanisms have been investigated, the effects of CO on cardiovascular function are still poorly understood. METHODS AND RESULTS: The effects of CO (250 ppm; 90 min) inhalation on myocardial function were investigated in isolated heart of rats killed immediately, and 3, 24, 48, and 96 h after CO exposure. CO exposure at 250 ppm resulted in an arterial carboxyhemoglobin (HbCO) level of approximately 11%, which was not associated with changes in mean arterial pressure and heart rate. CO exposure induced coronary perfusion pressure increases, which were associated with endothelium dependent and -independent vascular relaxation abnormalities. CO-induced coronary vascular relaxation perturbations were observed in the presence of increased heart contractility. Spontaneous peak to maximal Ca(2+)-activated left ventricular pressure ratio was markedly increased in CO-exposed rats, indicating increases in myofilament calcium sensitivity. Heart cyclic guanosine monophosphate/cAMP ratio and myocardial permeabilized fiber respiration (complex intravenous activity) were reduced in CO-exposed rats, which lasted after 48 h of reoxygenation in air. CONCLUSIONS: These findings suggest that CO deteriorates heart oxygen supply to utilization and potentially may induce myocardial hypoxia through mechanisms that include increased oxygen demand due to increased contractility, reduced coronary blood flow reserve, and cardiomyocyte respiration inhibition. PMID- 16690980 TI - Polymorphism within the interferon-gamma/receptor complex is associated with pulmonary tuberculosis. AB - RATIONALE: Interferon-gamma (IFN-gamma) is of central interest in the study of tuberculosis. A number of single-gene mutations have been identified in the IFN gamma signaling pathway that predispose to severe mycobacterial disease, but the relevance of polymorphism within these genes to the common phenotype of tuberculosis remains unclear. METHODS: A total of 1,301 individuals were included in a large, detailed study of West African populations with pulmonary tuberculosis. We investigated disease association with the genes encoding IFN gamma and its receptor subunits (IFNG, IFNGR1, and IFNGR2). RESULTS: Within the IFNG gene, two promoter variants showed evidence of novel disease association: 1616GG (odds ratio [OR], 1.49; 95% confidence interval [CI], 1.11-2.00; p = 0.008) and +3234TT (OR, 1.40; 95% CI, 1.09-1.80; p = 0.009). The +874AA genotype was not significantly more frequent among cases over control subjects (OR, 1.16; 95%CI, 0.89-1.51; p = 0.25). In addition, novel disease association was also found with the -56CC genotype of the IFNGR1 promoter (OR, 0.75; 95% CI, 0.57 0.99; p = 0.041). No disease association was seen with the IFNGR2 locus. CONCLUSIONS: These results provide evidence of a significant role for genetic variation at the IFNG locus and provide detailed understanding of the genetic mechanisms underlying this association. The disease association with IFNGR1 is novel, and together these findings support the hypothesis that genetically determined variation in both IFN-gamma production and responsiveness influences the risk of developing tuberculosis. PMID- 16690981 TI - Temperature curve complexity predicts survival in critically ill patients. AB - RATIONALE: Temperature curve complexity is inversely related to clinical status in critically ill patients. OBJECTIVE: To study if temperature curve complexity analysis predicts clinical outcome and how this test compares to other well established conventional measures. METHODS: Temperature was continuously recorded in 50 patients with multiple organ failure. Time-series complexity was analyzed using hourly approximate entropy (ApEn) and detrended fluctuation analysis (DFA) values. Sequential Organ Failure Assessment (SOFA) score was obtained every other day, and correlation between complexity and SOFA values was evaluated. Differences in complexity between nonsurviving and surviving patients were likewise analyzed. Logistic regression models were calculated to predict outcome, and receiver operating characteristic (ROC) curves were plotted to compare the predictive power of complexity values versus SOFA. MEASUREMENTS AND RESULTS: There was good correlation between complexity results and clinical scores for each patient. Nonsurvivors exhibited lower complexity values than survivors (minimum ApEn = 0.230 vs. 0.378; maximum DFA = 1.636 vs. 1.507; mean ApEn = 0.459 vs. 0.596; mean DFA = 1.376 vs. 1.288; p < 0.001 for all comparisons). In the logistic regression model, a change of 0.1 in the minimum complexity resulted in severe increases in the odds ratio of dying (7.6-fold for ApEn, 5.4-fold for DFA). In terms of predicting outcome, there were no significant differences in the areas under the ROC curves for complexity values versus SOFA scores. CONCLUSIONS: Low levels of complexity in the temperature curve are indicators of poor prognosis in patients with multiple organ failure. The predictive ability of temperature curve complexity is similar to that of the SOFA score. PMID- 16690982 TI - Reversibility of lung collapse and hypoxemia in early acute respiratory distress syndrome. AB - RATIONALE: The hypothesis that lung collapse is detrimental during the acute respiratory distress syndrome is still debatable. One of the difficulties is the lack of an efficient maneuver to minimize it. OBJECTIVES: To test if a bedside recruitment strategy, capable of reversing hypoxemia and collapse in > 95% of lung units, is clinically applicable in early acute respiratory distress syndrome. METHODS: Prospective assessment of a stepwise maximum-recruitment strategy using multislice computed tomography and continuous blood-gas hemodynamic monitoring. MEASUREMENTS AND MAIN RESULTS: Twenty-six patients received sequential increments in inspiratory airway pressures, in 5 cm H(2)O steps, until the detection of Pa(O(2)) + Pa(CO(2)) >or= 400 mm Hg. Whenever this primary target was not met, despite inspiratory pressures reaching 60 cm H(2)O, the maneuver was considered incomplete. If there was hemodynamic deterioration or barotrauma, the maneuver was to be interrupted. Late assessment of recruitment efficacy was performed by computed tomography (9 patients) or by online continuous monitoring in the intensive care unit (15 patients) up to 6 h. It was possible to open the lung and to keep the lung open in the majority (24/26) of patients, at the expense of transient hemodynamic effects and hypercapnia but without major clinical consequences. No barotrauma directly associated with the maneuver was detected. There was a strong and inverse relationship between arterial oxygenation and percentage of collapsed lung mass (R = - 0.91; p < 0.0001). CONCLUSIONS: It is often possible to reverse hypoxemia and fully recruit the lung in early acute respiratory distress syndrome. Due to transient side effects, the required maneuver still awaits further evaluation before routine clinical application. PMID- 16690983 TI - Asthma: is it due to an abnormal airway smooth muscle cell? AB - Asthma is an airway disease highly prevalent in westernized countries and of unknown etiology. Often, asthma is associated with atopy, but not all atopic individuals have asthma. Some patients with asthma outgrow symptoms, whereas many others acquire asthma later in life. Still other patients suffer from asthma their entire life. How can we explain these different patterns? It may be that asthma should be regarded as the clinical manifestation of a group of diseases with similar pathology due to a common factor. In this Pulmonary Perspective, we propose that an aberrant phenotype of airway smooth muscle (ASM) cells could be sufficient to explain the pathology of asthma. We will argue an abnormal ASM cell is a prerequisite to the development of asthma. Our postulate is that inadequate levels of C/EBPalpha, a protein that is pivotal for the suppression of both inflammation and proliferation responses, confer on ASM cells an activated phenotype that is more susceptible to mitogenic and contractile stimuli. PMID- 16690984 TI - Increased protein arginine methylation in chronic hypoxia: role of protein arginine methyltransferases. AB - Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide synthesis. ADMA is generated by catabolism of proteins containing methylated arginine residues, and its levels are correlated with endothelial dysfunction in systemic cardiovascular diseases. Arginine methylation of cellular proteins is catalyzed by protein arginine methyltransferases (PRMT). The expression and localization of PRMT in the lung has not been addressed. Here, we sought to analyze the expression of PRMT isoforms in the lung and to determine whether PRMT expression is altered during exposure to chronic hypoxia (10% oxygen). Adult mice were exposed to hypoxia for up to 3 wk, and lung tissues were harvested and processed for RT-PCR, Western blotting, immunohistochemistry, and determination of tissue ADMA levels. All PRMT isoforms investigated were detected at the mRNA and protein level in mouse lung, and were localized primarily to the bronchial and alveolar epithelium. In lungs of mice subjected to chronic hypoxia, PRMT2 mRNA and protein levels were up-regulated, whereas the expression of all other PRMT isoforms remained unchanged. This was mainly due to increased expression of PRMT2 in alveolar type II cells, which did not express detectable levels of PRMT2 under normoxic conditions. Consistent with these observations, lung ADMA levels and ADMA/l-Arginine ratios were increased under hypoxic conditions. These results demonstrate that PRMTs are expressed and functional in the lung, and that hypoxia is a potent regulator of PRMT2 expression and lung ADMA concentrations. These data suggest that structural and functional changes caused by hypoxia may be linked to ADMA metabolism. PMID- 16690985 TI - Expression of JP-8-induced inflammatory genes in AEII cells is mediated by NF kappaB and PARP-1. AB - Lung epithelial cells are critical in the regulation of airway inflammation in response to environmental pollutants. Altered activation of NF-kappaB is associated with expression of several proinflammatory factors in respiratory epithelial cells in response to an insult. Here we show that a low threshold dose (8 microg/ml) of the jet fuel JP-8 induces in a rat alveolar epithelial cell line (RLE-6TN) a prolonged activation of NF-kappaB as well as the increased expression of the proinflammatory cytokines TNF-alpha and IL-8, which are regulated by NF kappaB. The up-regulation of IL-6 mRNA in cells exposed to JP-8 appears to be a reaction of RLE-6TN cells to reduce the enhancement of proinflammatory mediators in response to the fuel. Moreover, lung tissues from rats exposed to occupational levels of JP-8 by nasal aerosol also showed dysregulated expression of TNF-alpha, IL-8, and IL-6, confirming the in vitro data. The poly(ADP-ribosyl)ation of PARP 1, a coactivator of NF-kappaB, was coincident with the prolonged activation of NF kappaB during JP-8 treatment. These results evidenced that a persistent exposure of the airway epithelium to aromatic hydrocarbons may have deleterious effects on pulmonary function. PMID- 16690986 TI - Neutrophil-derived elastase induces TGF-beta1 secretion in human airway smooth muscle via NF-kappaB pathway. AB - Neutrophils are infiltrated in airways of individuals with more severe and chronic asthma, with uncertain significance. Airway smooth muscle (ASM), apart from its contractile properties, is critically involved in the pathogenesis of asthma by producing inflammatory mediators. In the present study, we investigated the impact of neutrophil-derived elastase (NE) on ASM in terms of TGF-beta1 release, and we explored the underlying mechanisms. Primary ASM cells were serum starved for 24 h before stimulation with NE (0.01-0.5 microg/ml). TGF-beta1 in supernatant was determined by ELISA and mRNA quantified by real-time RT-QPCR. NF kappaB nuclear translocation and activation was examined by Western blotting and kappaB-2 dEGFP reporter gene assay. Association of IL-1 receptor-associated kinase (IRAK) with MyD88 was studied by co-immunoprecipitation and Toll-like receptor 4 (TLR4) determined by FACS scan and Western blotting. We demonstrated that NE enhanced TGF-beta1 release in a time-dependent manner. This induction was inhibited by actinomycin D (5 mM), cycloheximide (5 mM), and NF-kappaB inhibitors, including pyrrolidine dithiocarbamate (PDTC, 1 mM), aspirin (2.5 mM), and sodium salyicylate (2.5 mM). Stimulation with NE was rapidly followed by association of IRAK with MyD88, phosphorylation of IkappaBalpha, and nuclear translocation of p65 with increased transactivation activity. We also found that TLR4 levels were reduced upon NE treatment. These data suggest that NE upregulates TGF-beta1 gene expression and release via My88/IRAK/NF-kappaB pathway, possibly through activation of TLR4, and shed light on a potential role of neutrophils in the pathogenesis of asthma. PMID- 16690987 TI - Strain differences influence murine pulmonary responses to Stachybotrys chartarum. AB - When the fungus Stachybotrys chartarum is inhaled, its mycotoxins may cause lung injury and inflammation. The severity of human responses to S. chartarum in both occupational and home settings varies widely. To explore these differences, we intratracheally instilled C3H/HeJ, BALB/c, and C57BL/6J mice with S. chartarum spores suspended in saline. One day later, the mice were humanely killed, bronchoalveolar lavage (BAL) was performed, and biochemical and cellular indicators of lung injury and inflammation were measured. BALB/c mice showed the highest myeloperoxidase activity, albumin and hemoglobin levels, and neutrophil numbers in their BAL among the three strains. BALB/c was the only strain to show significant increases in keratinocyte-derived cytokine (KC), monocyte chemotactic protein (MCP)-1, MCP-3, macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, MIP-1gamma, MIP-2, RANTES, IL-1alpha, IL-1beta, IL-3, IL-6, IL-18, leukemia inhibitory factor, macrophage colony-stimulating factor, and TNF-alpha. A model of allergen-induced airway inflammation was examined to assess whether underlying allergic inflammation might contribute to increased susceptibility to S. chartarum-induced pulmonary inflammation and injury. Surprisingly, in BALB/c mice, ovalbumin-induced airway inflammation produced a protective effect against some S. chartarum-induced pulmonary responses. This is the first report of mammalian strain differences affecting responses to S. chartarum. These responses differ from those reported for LPS and other fungi. Analogous underlying genetic differences may contribute to the wide range of sensitivity to Stachybotrys among humans. PMID- 16690989 TI - Patterns of cognitive performance in middle-aged and older adults: A cluster analytic examination. AB - Cognitive decline in speeded abilities, executive function, and memory is believed to typify normal aging. However, there is significant variability in cognitive function with advanced age and some reports of relatively intact cognitive function among a subset of older individuals. The present study consists of a cluster analysis to examine the patterns of cognitive function in middle-aged and older individuals. Analyses revealed 3 clusters of middle-aged adults, including an intact group, persons with poor motor speed, and a group with reduced executive function. Three clusters were also identified for older adults, including a group with poor executive function, persons with reduced speed performance (attention, executive function, motor), and a group with global cognitive decline. No evidence emerged for a cluster of older adults with intact performance in all domains or with isolated memory deficits. Findings generally support the frontal aging hypothesis and may provide important information about healthy cognitive aging. PMID- 16690988 TI - Gene expression profile of human airway epithelium induced by hyperoxia in vivo. AB - Hyperoxia leads to oxidative modification and damage of macromolecules in the respiratory tract with loss of biological functions. Given the lack of antioxidant gene induction with acute exposure to 100% oxygen, we hypothesized that clearance pathways for oxidatively modified proteins may be induced and serve in the immediate cellular response to preserve the epithelial layer. To test this, airway epithelial cells were obtained from individuals under ambient oxygen conditions and after breathing 100% oxygen for 12 h. Gene expression profiling identified induction of genes in the chaperone and proteasome-ubiquitin conjugation pathways that together comprise an integrated cellular response to manage and degrade damaged proteins. Analyses also revealed gene expression changes associated with oxidoreductase function, cell cycle regulation, and ATP synthesis. Increased HSP70, protein ubiquitination, and intracellular ATP were validated in cells exposed to hyperoxia in vitro. Inhibition of proteasomal degradation revealed the importance of accelerated protein catabolism for energy production of cells exposed to hyperoxia. Thus, the human airway early response to hyperoxia relies predominantly upon induction of cytoprotective chaperones and the ubiquitin-proteasome-dependent protein degradation system to maintain airway homeostatic integrity. PMID- 16690990 TI - Frequency and clinical determinants of poststroke cognitive impairment in nondemented stroke patients. AB - The objective of this study was to examine the prevalence and clinical correlates of poststroke cognitive impairment in Chinese stroke patients in Hong Kong. One hundred seventy-nine stroke patients were interviewed by a psychiatrist 3 months after their stroke. Cognitive impairment was determined according to the Mini Mental State Examination score. Thirty-nine participants (21.8%) had cognitive impairment. Univariate analysis found that cognitive impairment was associated with age, female sex, level of education, previous stroke, prestroke Rankin score, National Institutes of Health Stroke Scale dysarthria and total scores, urinary incontinence, and cerebral atrophy index. Multivariate logistic regression suggested that female sex, education, National Institutes of Health Stroke Scale dysarthria score, urinary incontinence, and atrial fibrillation were independent risk factors of poststroke cognitive impairment. After removal of 54 patients with previous stroke from the sample, the frequency of cognitive impairment decreased to 18.4%. It was concluded that cognitive impairment is common among nondemented Chinese stroke patients in Hong Kong. PMID- 16690991 TI - Factors associated with psychiatric hospitalization of individuals diagnosed with dementia and comorbid bipolar disorder. AB - The objective was to determine risk factors of psychiatric hospitalization among a Veterans Administration database of patients with dementia and comorbid bipolar disorder (D+BD). Patients with D+BD had a greater prevalence of psychiatric hospitalization (28% vs 4%). The strongest predictor of psychiatric hospitalization was the presence of an alcohol use disorder (51% risk); patients without alcohol use disorders but under the age of 70 had the next highest risk (33% risk). However, patients with an alcohol use disorder had shorter psychiatric hospitalizations than those without. Compared with patients without BD, D+BD patients were more likely to have alcohol use disorders (15% vs 3%) and any other substance use problem (10% vs 1%). In patients diagnosed with dementia and bipolar disorder, the strongest risk factor for psychiatric hospitalization was an alcohol abuse disorder. These findings suggest that disorders with increased frequency in BD affect the course of dementia. PMID- 16690992 TI - Vascular disease and risk factors, rate of progression, and survival in Alzheimer's disease. AB - Two hundred forty-seven patients with early Alzheimer's disease were studied for the association of demographic, functional, and cognitive status and vascular comorbidities and risk factors present at index visit to rate of clinical disease progression over 3 years and to survival time. Patients who progressed to the moderate stage were designated fast progressors; those who remained in the early stage were designated slow progressors. At index visit, Mini-Mental State Exam score was significantly lower for the fast than the slow group; global impairment was significantly higher for the fast group. Cognitive scores showed greater annual decline in the fast group, and the fast group also had a greater annualized global change. The fast group had a shorter median survival time from onset, but age at onset, age at initial visit, history of heart problems, myocardial infarct, stroke, hypertension, diabetes, or past or current smoking did not differ between groups. PMID- 16690993 TI - Delirium in older patients admitted to general internal medicine. AB - Delirium on the day of admission to general internal medicine wards was studied in 400 consecutive patients aged 70 years and above regarding occurrence, associated factors, clinical profile, length of hospital stay, and mortality. The patients were assessed using the Organic Brain Syndrome Scale and the Mini-Mental State Examination, and delirium was diagnosed according to Diagnostic and Statistical Manual of Mental Disorders (4th ed) criteria. Delirium on the day of admission occurred in 31.3% of the patients and was independently associated with old age, fever on the day of admission (> or = 38 degrees C), treatment with neuroleptics, impaired vision, male sex, and previous stroke. Delirious patients had longer hospital stay (15.4 vs 9.5 days, P < .001), a higher mortality rate during hospitalization (11/125 vs 5/275, P < .001), and a higher 1-year mortality rate (45/125 vs 55/275, P = .001). Delirium is a common complication with often easily identified causes, and it has a serious impact on outcome for older medical patients. PMID- 16690994 TI - Patterns of initial pharmacotherapy for Parkinson's disease in the United States. AB - Data from a mix of employer- and government-funded health plans were used to investigate actual treatment patterns for patients initiating pharmacotherapy for Parkinson's disease in the United States. Treatment patterns evaluated included type of initial therapy and rates and types of adjunctive and substitute therapies. The study confirms that levodopa remains the most often prescribed initial treatment for Parkinson's disease regardless of age or drug benefit coverage. The widespread use of levodopa in young Parkinson's patients (<65 years) with private insurance may indicate that physicians are not overly concerned about or are not fully aware of the association of levodopa with long term motor complications. It may also indicate that currently available alternatives to levodopa are not sufficiently effective or well tolerated. PMID- 16690995 TI - Light to moderate alcohol consumption is associated with better cognitive function among older male veterans receiving primary care. AB - Among older persons, the effects of light to moderate alcohol consumption on cognitive function remain inadequately defined. The authors sought to determine whether light to moderate drinking is associated with better cognitive function among older men. Participants included men aged 65 years or older enrolled in a Veteran's Administration (VA) primary care clinic. Current (past 1 year) and lifetime use, cognitive functioning (as determined by the Trail Making Part B, Symbol Digit, FAS, and Hopkins Verbal Learning tests), and demographic, psychosocial, and medical status were obtained using standardized methods. Participants (N = 760) had a mean age of 74 (range, 65-89) years. Current drinkers (n = 509) as compared with never (n = 31) and former (n = 220) drinkers demonstrated significantly better cognitive performance on 3 (Trails B, Symbol Digit, and Hopkins Verbal Learning) of the 4 tests (P < .01 for all comparisons). In multiple linear regression models, current light to moderate drinking (ie, 7 or fewer drinks per week), as compared to a reference group of never and former drinkers, was associated with better performance on the Trails B, Symbol Digit, and Hopkins Verbal Learning tests (P < .01 for all comparisons). The number of years drinking 7 or fewer drinks per week also was independently associated with better cognitive performance. Current consumption of 7 or fewer drinks per week and the number of years drinking at this level are both associated with better cognitive performance in older male veterans receiving primary care. These findings are consistent with the hypothesis that light to moderate drinking confers cognitive benefits to older persons. PMID- 16690997 TI - IL-15 is elevated in cerebrospinal fluid of patients with Alzheimer's disease and frontotemporal dementia. AB - Interleukin-15 is a novel proinflammatory cytokine. It is produced by activated blood monocytes, macrophages, and glial cells. The objective of our study was to assess the role of interleukin-15 as a marker of increased proinflammatory activity in patients with Alzheimer's disease and frontotemporal dementia. We measured cerebrospinal fluid interleukin-15 levels in 17 patients with Alzheimer's disease and 7 patients with frontotemporal dementia in comparison with 17 patients with amyotrophic lateral sclerosis and 15 patients with Parkinson's disease. Patients with Alzheimer's disease and frontotemporal dementia had significantly higher cerebrospinal fluid interleukin-15 levels compared with patients with noninflammatory neurological diseases (P < .05 and P < .01, respectively). In Alzheimer's disease, a significant positive correlation was noted between interleukin-15 levels and age of onset (R = .48, P = .05). Our findings suggest that interleukin-15 may be implicated in the pathophysiology of Alzheimer's disease and frontotemporal dementia. PMID- 16690996 TI - Severity of gambling problems and psychosocial functioning in older adults. AB - This study examined the relationship between severity of gambling problems and psychosocial functioning in older adults. Twenty-one current pathological gamblers and 10 current problem gamblers over 60 years of age completed the Addiction Severity Index (ASI), Geriatric Depression Scale (GDS), Brief Symptom Inventory (BSI), UCLA Loneliness Scale (UCLA-LS), and Social Provisions Scale (SPS). Compared with problem gamblers, pathological gamblers reported increased severity of gambling and family/social problems on the ASI; scored higher on the GDS, BSI, and UCLA-LS; and scored lower on the SPS. Although they were not seeking treatment at the time of the interview, three fourths of pathological gamblers and 30% of problem gamblers were interested in gambling treatment. These results suggest that severity of gambling problems is associated with increased psychosocial distress in older adults and that a significant proportion of older adults with a gambling disorder may be cognizant of their gambling problem and interested in treatment. PMID- 16690998 TI - Chemical rescue, multiple ionizable groups, and general acid-base catalysis in the HDV genomic ribozyme. AB - In the ribozyme from the hepatitis delta virus (HDV) genomic strand RNA, a cytosine side chain is proposed to facilitate proton transfer in the transition state of the reaction and, thus, act as a general acid-base catalyst. Mutation of this active-site cytosine (C75) reduced RNA cleavage rates by as much as one million-fold, but addition of exogenous cytosine and certain nucleobase or imidazole analogs can partially rescue activity in these mutants. However, pH rate profiles for the rescued reactions were bell shaped, and only one leg of the pH-rate curve could be attributed to ionization of the exogenous nucleobase or buffer. When a second potential ionizable nucleobase (C41) was removed, one leg of the bell-shaped curve was eliminated in the chemical-rescue reaction. With this construct, the apparent pK(a) determined from the pH-rate profile correlated with the solution pK(a) of the buffer, and the contribution of the buffer to the rate enhancement could be directly evaluated in a free-energy or Bronsted plot. The free-energy relationship between the acid dissociation constant of the buffer and the rate constant for cleavage (Bronsted value, beta, = approximately 0.5) was consistent with a mechanism in which the buffer acted as a general acid-base catalyst. These data support the hypothesis that cytosine 75, in the intact ribozyme, acts as a general acid-base catalyst. PMID- 16690999 TI - RNA editing complex interactions with a site for full-round U deletion in Trypanosoma brucei. AB - Trypanosome U insertion and U deletion RNA editing of mitochondrial pre-mRNAs is catalyzed by multisubunit editing complexes as directed by partially complementary guide RNAs. The basic enzymatic activities and protein composition of these high-molecular mass complexes have been under intense study, but their specific protein interactions with functional pre-mRNA/gRNA substrates remains unknown. We show that editing complexes purified through extensive ion-exchange chromatography and immunoprecipitation make specific cross-linking interactions with A6 pre-mRNA containing a single 32P and photoreactive 4-thioU at the scissile bond of a functional site for full-round U deletion. At least four direct protein-RNA contacts are detected at this site by cross-linking. All four interactions are stimulated by unpaired residues just 5' of the pre-mRNA/gRNA anchor duplex, but strongly inhibited by pairing of the editing site region. Furthermore, competition analysis with homologous and heterologous transcripts suggests preferential contacts of the editing complex with the mRNA/gRNA duplex substrate. This apparent structural selectivity suggests that the RNA-protein interactions we observe may be involved in recognition of editing sites and/or catalysis in assembled complexes. PMID- 16691000 TI - RBP16 stimulates trypanosome RNA editing in vitro at an early step in the editing reaction. AB - RBP16 is an abundant RNA binding protein from Trypanosoma brucei mitochondria that affects both RNA editing and stability. We report here experiments aimed at elucidating the mechanism of RBP16 function in RNA editing. In in vitro RNA editing assays, recombinant RBP16 is able to significantly stimulate insertion editing of both CYb and A6 pre-mRNAs. Enhancement of in vitro editing activity occurs at, or prior to, the step of pre-mRNA cleavage, as evidenced by increased accumulation of pre-mRNA 3' cleavage products in the presence of RBP16. Mutated RBP16 that is severely compromised in cold shock domain (CSD)-mediated RNA binding was able to enhance editing to levels comparable to the wild-type protein in some assays at the highest RBP16 levels tested. However, at low RBP16 concentrations or in assays with native, oligo(U)-tail-bearing gRNAs, editing stimulation by mutant RBP16 was somewhat compromised. Together, these results indicate that both the N-terminal CSD and C-terminal RGG RNA binding domains of RBP16 are required for maximal editing stimulation. Finally, the relaxed specificity of RBP16 for stimulation of both CYb and A6 editing in vitro implicates additional specificity factors that account for the strict CYb specificity of RBP16 action in editing in vivo. Our results constitute the first report of any putative RNA editing accessory factor eliciting an effect on editing in vitro. Overall, these results support a novel accessory role for RBP16 in U insertion editing. PMID- 16691001 TI - Noncovalently linked nuclear localization peptides for enhanced calcium phosphate transfection. AB - The generation of cell lines stably expressing recombinant material is a lengthy process and there has thus been much interest in the use of transient expression systems to rapidly produce recombinant material. To achieve this, the DNA of interest must be delivered into the nucleus of the target cell. The mechanisms by which this process occurs are poorly understood and the efficiency of various methods differs widely. Recently, nuclear localization signals (NLSs) have been investigated to target entry of DNA into the nucleus of mammalian cells. We have used NLSs from the SV40 and Tat antigens mixed with our model luciferase reporter gene plasmid for the transfection of Chinese hamster ovary (CHO) cells using calcium phosphate and FuGENE 6 transfection technology. The noncovalent complexation of NLSs with plasmid DNA before calcium phosphate-mediated transfection resulted in enhanced reporter gene expression with increasing ratios of NLS to plasmid until reaching a maximum. At higher ratios than maximum expression, the expression levels decreased. On the other hand, when using FuGENE 6 reagent NLSs did not enhance reporter gene expression. Cell cycle arrest in G(2)/M phase obliterated the effect of the NLS on reporter gene expression when using the calcium phosphate transfection method. PMID- 16691002 TI - An inducible T7 RNA polymerase-dependent plasmid system. AB - RNA interference (RNAi) has become a powerful tool for the specific silencing of gene transcription. Especially the targeting of genes in mammalian cells has been greatly improved by generating plasmid based and viral vector-based systems. This permits expression of short hairpin RNA (shRNA) on a longterm basis. However, an inducible expression of shRNA is required, if the target is essential for cell survival. We developed a doxycycline-inducible two-plasmid system for the expression of a ribozyme-processed shRNA. In contrast to other existing systems, we use the highly specific T7 phage RNA polymerase, which does not interact with cellular factors; therefore, interference with cellular functions is limited. One plasmid is responsible for doxycycline-dependent expression of T7 RNA polymerase and a second plasmid expresses a ribozyme-processed shRNA under the control of a T7 promoter. Our results showed that doxycycline- dependent expression of T7 RNA polymerase was tightly controlled and expression of an shRNA against firefly luciferase inhibited 86% of luciferase activity. In conclusion, our plasmid system provides a very useful tool for analyzing essential gene functions in vitro. PMID- 16691003 TI - A universal cloning vector using vaccinia topoisomerase I. AB - Vaccinia DNA topoisomerase I (TOPO) charged vectors with a sticky T are routinely used to clone polymerase chain reaction (PCR) products with an extra A at their 3' end (TOPO TA Cloning from Invitrogen). TOPO charged blunt vectors are used to clone blunt end PCR products (TOPO Blunt Cloning). Here, we demonstrate that both TOPO TA vectors and TOPO Blunt vectors can be used to clone PCR products with either a blunt end or an extra A at the 3' end. We further demonstrate that these vectors can be used to clone sticky end DNA generated with restriction enzymes. In summary, these TOPO vectors can be used as universal cloning vectors. PMID- 16691004 TI - Cloning, expression, and purification of lipoprotein-associated phospholipase A(2) in Pichia pastoris. AB - Lipoprotein-associated phospholipase A(2) (Lp-PLA(2)) has been shown to play a crucial role in atherosclerosis, and has been proposed as a promising target for drug discovery. Here, we cloned the Lp-PLA(2) gene from differentiated THP-1 cells, and inserted a carboxy-terminal His(6)-tagged version of the gene into the pPIC9 Pichia expression vector. The Lp-PLA(2) fusion protein was successfully expressed in Pichia pastoris expression system and could be rapidly purified to apparent homogeneity using a single-step purification method. The activity of our recombinant Lp-PLA(2) was strong when [3H] PAF was used as a substrate, and the Lp-PLA(2) inhibitor SB435495 exhibited an inhibitory curve against the recombinant Lp-PLA2 (IC50 = 15.93 +/- 1 microM). This novel recombinant Lp-PLA(2) could prove useful as a screening model for Lp-PLA(2) inhibitors, and may facilitate further investigation of this protein in atherosclerosis. PMID- 16691005 TI - The use of unilateral PCR to identify prominent heteroduplexes formed during PCR of the mouse microsatellite locus D17Mit23. AB - Microsatellite markers are useful tools for understanding the evolutionary history of discrete segments of the mammalian genome. We used the microsatellite marker D17Mit23 to study the portion of the mouse genome known as the t complex, a naturally occurring variant of Chromosome 17. We identified an allelic variant of D17Mit23, which is shared by two forms of the t complex, the t haplotypes t(w2) and t(Lub2). Polymerase chain reaction (PCR) analysis of DNA samples from mice that were heterozygous for either t haplotype resulted in gel patterns with prominent bands of higher molecular weight in addition to the bona-fide D17Mit23 alleles. The appearance of these higher molecular weight bands, although consistent with heteroduplex formation, was not diminished through the use of reconditioning PCR. We used a modified form of asymmetric PCR, called "unilateral PCR," to show that the higher molecular weight bands are heteroduplexes and to identify their constituent strands. Certain microsatellite motifs may be especially prone to the production of prominent heteroduplex products, and this may lead to the erroneous genotyping of DNA samples. PMID- 16691006 TI - An extraordinarily high level of IL-15 expression by a cell line transduced with a modified BMGneo vector displays hypoxic upregulation. AB - Interleukin (IL)-15 expression level is tightly controlled in mammalian cells by various mechanisms. In order to achieve higher expression levels of IL-15, many attempts have been made, but the highest expression rate among those reported is still only 13.3 ng/106 cells/24 h. Here we report that a selected human embryonic kidney 293 (HEK293) cell line, denoted 293HAN cells, which can survive and proliferate under conditions of hypoxia, acidity, and nutritional depletion (HAN), after transduction -- with a modified BMGneo vector -- can produce functional human IL-15 at the extremely high rate of 890 ng/106 cells/24 h under normoxic conditions -- a 67-fold increase. This is as a result of multiple episomally based vector copy numbers per cell. An extra benefit was that the BMGneo vector was found to be inducible in hypoxia and allowed a further approximately threefold upregulation of the human IL-15 level which made these 293HAN cells, transduced with the modified BMGneo vector, a very promising tool for high IL-15 production (approximately 200-fold increase above that of baseline normoxia). The mechanism of hypoxic upregulation was found to be related to the mouse MT-1 promoter present in the vector. PMID- 16691007 TI - Probing the antigenicity of E. coli L-asparaginase by mutational analysis. AB - A strategy, termed alanine-scanning mutagenesis, was used to identify the amino acid residues which are critical to the antigenicity of Escherichia coli L asparaginase (L-ASP). Three continuous alkaline residues, 195RKH197, were mutated to Ala selectively. Four mutant recombinant L-ASPs were constructed and expressed in E. coli, and then purified. The purified mutants showed a single band by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and were more than 95% pure by reverse high-performance liquid chromatography. The activities of wild type and mL-ASPs in the fermentative medium were all about 130 U/mL. The change from 195RKH197 to 195AAA197 reduced the antigenicity of the enzyme greatly as shown in competition enzyme-linked immunosorbent assay using polyclonal antibodies raised against the wild-type L-ASP from rabbits. The results show that residues 195RKH197 of E. coli L-ASP are critical to its antigenicity. PMID- 16691008 TI - A rapid and simple method for DNA extraction from yeasts and fungi isolated from Agave fourcroydes. AB - A simple and easy protocol for extracting high-quality DNA from different yeast and filamentous fungal species is described. This method involves two important steps: first, the disruption of cell walls by mechanical means and freezing; and second, the extraction, isolation, and precipitation of genomic DNA. The absorbance ratios (A(260)/A(280)) obtained ranged from 1.6 to 2.0. The main objective of this procedure is to extract pure DNA from yeast and filamentous fungi, including those with high contents of proteins, polysaccharides, and other complex compounds in their cell walls. The yield and quality of the DNAs obtained were suitable for micro/minisatellite primer-polymerase chain reaction (MSP-PCR) fingerprinting as well as for the sequence of the D1/D2 domain of the 26S rDNA. PMID- 16691009 TI - Molecular aspects of disease pathogenesis in the transmissible spongiform encephalopathies. AB - The transmissible spongiform encephalopathies (TSE), or prion diseases, are a group of rare, fatal, and transmissible neurodegenerative diseases of mammals for which there are no known viral or bacterial etiological agents. The bovine form of these diseases, bovine spongiform encephalopathy (BSE), has crossed over into humans to cause variant Creutzfeldt-Jakob disease. As a result, BSE and the TSE diseases are now considered a significant threat to human health. Understanding the basic mechanisms of TSE pathogenesis is essential for the development of effective TSE diagnostic tests and anti-TSE therapeutic regimens. This review provides an overview of the molecular mechanisms that underlie this enigmatic group of diseases. PMID- 16691010 TI - Bombesin/gastrin-releasing peptide receptor antagonists increase the ability of histone deacetylase inhibitors to reduce lung cancer proliferation. AB - The effects of a bombesin/gastrin releasing peptide (BB/GRP) receptor antagonist, PD176252, and histone deacetylase (HDAC) inhibitor, MS-275, were investigated on human lung cancer cell lines. Using the MTT assay, PD176252 and MS-275 inhibited the proliferation of NCI-H1299 cells with IC50 values of 7 and 5 microg/mL, respectively. Using MS-275 and PD176252 together, the ability to inhibit lung cancer cellular growth increased significantly. The combination index for MS-275 and PD176252 was <0.2, indicating that the compounds are highly synergistic in inhibiting lung cancer cellular growth. Also, MS-275 and PD176252 together strongly inhibited the clonal growth of NCI-H345 or NCI-H1299 cells. MS-275 had little effect on the expression of lung cancer cellular GRP or GRP receptors, but increased expression of transforming growth factor-beta receptor II (TGF-beta RII). These results indicate that GRP receptor antagonists may potentiate the action of histone deacetylase inhibitors on lung cancer cellular proliferation by increasing expression of tumor suppressor genes. PMID- 16691011 TI - Emotional behavior in aged neuropeptide Y (NPY) Y2 knockout mice. AB - Neuropeptide Y (NPY) was shown to modulate anxiety- and depression-related behaviors in various animal models. Previous studies demonstrated that NPY Y2 receptor knockout (KO) mice display an anxiolytic- and antidepressant-like phenotype compared with control animals. However, the long-term effect of the deletion of this receptor in aged animals is unknown. Thus, anxiety- and depression-related behaviors were investigated in 2-yr-old NPY Y2 KO mice. Aged NPY Y2 KO mice display an anxiolytic-like profile as assessed in the elevated plus-maze and open field, providing further support for a role for Y2 receptors in anxiety-related behaviors. Furthermore, aged NPY Y2 KO mice have significantly lower immobility scores in the forced swim test; supporting the role for this receptor in antidepressant-like behaviors. These data provide further evidence that modulators of the NPY Y2 receptor subtype are drug targets for the treatment of anxiety and mood disorders in human subjects. PMID- 16691012 TI - ARP, the cleavable C-terminal peptide of "readthrough" acetylcholinesterase, promotes neuronal development and plasticity. AB - The mammalian acetylcholinesterase (ACHE) gene gives rise to diverse enzymatically active proteins with three different carboxyl termini. In the brain, the normally rare readthrough AChE-R monomer accumulates under embryonic development and in adults following psychological stress, head injury, or exposure to AChEs. In the prenatal developing cortex, its unique C-terminal peptide ARP associates with radial glial fibers supporting neuronal migration. In contrast, the major synaptic AChE-S variant appears in the migrating neurons themselves. Moreover, antisense suppression of AChE-R attenuates neuronal migration, allowing increased proliferation of neuronal progenitors. In the adult brain, neuronal AChE-R is either secreted or accumulates intraneuronally, where it interacts through ARP with the scaffold protein RACK1 and activated PKC betaII. This associates with increased PKC-betaII activity, which shuttles to submembranal clusters (e.g., in hyperactivated hippocampal neurons). Cleavage yields the AChE-R-specific C-terminal peptide, including immunopositive ARP. Importantly, intrahippocampal injection of synthetic ARP was followed by its efficient neuronal penetration and retrograde transport into cortical and basal nuclei neurons. Moreover, ARP-injected mice presented increased stress-induced contextual fear, inhibitable by antisense suppression of AChE-R mRNA. Together, our findings point at the cleavable ARP peptide as a key regulator of neuronal development and plasticity and suggest its use as a drug target and/or research and therapeutic tool. PMID- 16691013 TI - Retinal growth hormone in perinatal and adult rats. AB - Growth hormone (GH) mRNA and protein have recently been localized in the neural retina of embryonic chicks, in which exogenous GH promotes cell survival. GH is also expressed in the rat CNS, in which it has neuroprotective roles, although its presence in the rat neural retina is unknown and is the focus of the present study. GH immunoreactivity, to a 22-kDa protein, was present in extracts of fetal (embryonic day [ED]17) eyes and in extracts from the neural retinas of newborn pups, comparable to GH immunoreactivity in pituitary extracts. The GH immunoreactivity in the neural retina was widespread but was most intense in large rounded cells in the retinal ganglion cell (RGC) layer and in the optic fiber layer derived from the axons of the RGCs. A 693-bp cDNA was also generated by the RT-PCR of RNA extracted from the eyes of ED17 rats and from the neural retinas and eyes of newborn rats, when amplified in the presence of oligonucleotide primers for the rat GH cDNA. Expression of the GH gene in the neural retina was also shown by specific in situ hybridization of an antisense GH riboprobe to cells in the neural retina, particularly those in the RGC layers of fetal and adult rat eyes. These results demonstrate GH expression in the neural retinas of fetal, newborn, and adult rats, in which retinal GH might have neuroprotective roles. PMID- 16691014 TI - Quantitative peptidomics in mice: effect of cocaine treatment. AB - We recently developed a quantitative peptidomics method using stable isotopic labels and mass spectrometry to both quantify and identify a large number of peptides. To test this approach and screen for peptides regulated by cocaine administration, 32 Cpefat/fat mice and 16 wild-type mice were treated twice daily for 5 d either with saline or 10 mg/kg cocaine. Peptides were extracted from striatum, hypothalamus, hippocampus, and prefrontal cortex, and extracts from groups of eight mice were labeled with the N-hydroxysuccinimide ester of trimethylammonium butyrate containing either nine deuterium or nine hydrogen atoms. Pools of heavy- and light-labeled peptides were combined, purified on an anhydrotrypsin affinity column, and analyzed on a reversephase column coupled to an electrospray ionization quadrapole time-of-flight mass spectrometer. Changes in peptide levels upon cocaine treatment were determined from the relative peak intensities of the cocaine versus saline peaks, and peptides were identified from collision-induced dissociation spectra. Ten peptides were found to increase or decrease in each of two separate analyses from distinct groups of mice. Peptides found to increase corresponded to fragments of proenkephalin, prothyrotropin releasing hormone, provasopressin, proSAAS, secretogranin II, chromogranin B, and peptidyl-glycine-alpha-amidating mono-oxygenase in the hypothalamus. The same peptidyl-glycine-alpha-amidating mono-oxygenase peptide decreased in the prefrontal cortex, along with striatal neurokinin B and two unidentified peptides. Thirty other peptides were not substantially affected by cocaine treatment in both replicates. Taken together, the quantitative peptidomics approach provides an efficient method to screen for changes in a large number of peptides. PMID- 16691015 TI - Peptidomics of Cpefat/fat mouse hypothalamus and striatum: effect of chronic morphine administration. AB - Chronic morphine administration is known to affect several neuropeptide systems, and this could contribute to the behavioral effects of opiates. To quantitate global changes in neuropeptide levels upon chronic morphine administration, we took advantage of a method that allows selective isolation of neuropeptides from brains of mice lacking carboxypeptidase E (Cpefat/fat mice), a critical enzyme in the generation of many neuroendocrine peptides. We used a differential labeling procedure with stable isotopic tags and mass spectrometry to quantitate the relative changes in a number of hypothalamic and striatal peptides in Cpefat/fat mice chronically treated with morphine. A total of 27 distinct peptides were detected in hypothalamus and striatum. Of these, 27 were identified by mass spectrometry-based sequencing, 1 was tentatively identified by the mass and charge, and 9 were not identified. The identified peptides included fragments of proenkephalin, prothyrotropin-releasing hormone, secretogranin II, chromogranin Aand B, protachykinin B, provasopressin, promelanin concentrating hormone, and pro-SAAS. Upon morphine administration, although the levels of most of the peptides were unaltered (within a factor of 1.3 to 0.7 compared with saline control), the levels of a small number of peptides did show consistent changes (increased or decreased by 1.3-fold or more) in hypothalamus and/or striatum. Taken together, these results provide interesting insights into endogenous neuropeptide systems that are modulated by morphine and suggest further experiments to link candidate peptides with long-term effects of morphine. PMID- 16691016 TI - Angiotensin II type-1 receptor A1166C polymorphism is associated with increased risk of ischemic stroke in hypertensive smokers. AB - Recent observations revealed a novel role of angiotensin-converting enzyme 2 and the angiotensin II type-1 receptor (AT1R) in lung injury, thereby extending knowledge about the functions of the angiotensin system. Angiotensin II, whose target is the AT1R, is a potent vasoconstrictor. Accordingly, an imbalance leading to enhanced activity of the angiotensin II-AT1R axis is postulated to contribute to both circulatory disturbances and lung injury. In this context, a functional single-nucleotide polymorphism, AT1R A1166C, which leads to enhanced responsiveness of the AT1R, has been postulated as a candidate susceptibility factor for ischemic stroke. The aim of our study was to investigate its occurrence in ischemic stroke and to analyze its possible synergistic associations with clinical risk factors. Genetic and clinical data on 308 consecutive patients with acutely developing ischemic stroke were analyzed. A total of 272 stroke and neuroimaging alteration-free subjects served as a control group. Univariate and logistic regression statistical approaches were used. Alone, the AT1R 1166C allele did not pose a risk of stroke. In hypertensive smokers, however, it was associated with an increased risk of ischemic stroke (OR 22.3, 95% CI 5.8-110.2, p<0.001). Further subgroup analysis revealed the same association for both small-vessel (OR 24.3, 95% CI 6.1-121.1, p<0.001) and large vessel (OR 21.3, 95% CI 4.6-81.1, p<0.001) infarction. On a pathophysiological basis, our results suggest the possibility that the AT1R A1166C polymorphism might give rise to ischemic stroke indirectly via an unfavorable effect on the cardiorespiratory function. PMID- 16691017 TI - Effects on hippocampus of lifelong absence of glucocorticoids in the pro opiomelanocortin null mutant mouse reveal complex relationship between glucocorticoids and hippocampal structure and function. AB - In humans changes in serum cortisol levels have been observed with aging, stress, and with affective disorders such as major depression and post-traumatic stress disorder. Corticosteroids are known to influence hippocampal structure and function; specifically, plasma corticosteroid levels have been inversely correlated with hippocampal cell proliferation, cell death, and impaired memory function. The relationship between corticosteroids and structure and function of the hippocampus has been studied in experimental systems in adult animals by increasing or decreasing corticosterone levels through pharmacological supplementation and through surgical removal of the adrenal gland. Here, we utilized the genetically engineered pro-opiomelanocortin (POMC) null mutant mouse, which because of the lack of all POMC peptides has no corticosterone from birth throughout life. The effect of this lifelong absence of corticosterone on the dentate gyrus of the hippocampus is a decrease in granule cell density, which correlated with a decrease in cell proliferation but not an increase in cell degeneration. Fine morphology of granule cells was unaltered. Analyses of gene expression revealed no changes in POMC null mutant vs wild-type hippocampus with respect to levels of expression of corticoid receptor genes or genes known to be regulated by corticosterone. Spatial learning as tested by the Morris water maze was not altered in the POMC null mutant mouse. Taken together with findings from other studies of the effects of altered levels of corticosteroids on the hippocampus, our results argue for a complex homeostasis in which disturbances of any one factor can offset the system in varying ways. PMID- 16691018 TI - Tubulin is the target binding site for NAP-related peptides: ADNF-9, D-NAP, and D SAL. AB - The authors set out to investigate whether NAP-related peptides interact with tubulin at a NAP binding site. Previous studies have shown that the neuroprotective peptide NAP binds to tubulin. As NAP (NAPVSIPQ) shares structural similarities with ADNF-9 (SALLRSIPA), and the all-D-enantiomers, D-NAP and D-SAL, it was hypothesized that all of these peptides compete with NAP-tubulin binding. Using NAP affinity column and extracts from newborn rat brain (cerebral cortex), we now show that the above-mentioned peptides compete with NAP binding to tubulin. The identification of tubulin as a target binding site for NAP-related peptides explains, in part, the broad neuroprotective activity offered by these potent peptides. PMID- 16691019 TI - Vitamin B6 deficiency and dietary fats: effects on lipid composition and glutathione peroxidase activity in rat liver. AB - Dietary selenium, vitamin B6 and fatty acids modulate both tissue acyl composition by regulating polyunsaturated fatty acid metabolism and antioxidant defences by influencing glutathione peroxidase activity. Alteration in the intake of one of them could therefore lead to different results depending on the intake of the others. To clarify this complex relationship, in the present study we have evaluated the modifications occurring in fatty acid composition and glutathione peroxidase activity in total liver and liver microsomes of rats fed diets containing the same amount of selenium, but different vitamin B6 content and fatty acid composition. Our data indicate that both acyl composition and glutathione peroxidase activity are greatly influenced not only by vitamin B6 deficiency, but also by the diet unsaturation degree. This study underlines that not only selenium availability but also other nutrients can modulate glutathione peroxidase activity. PMID- 16691020 TI - High blood pressure and inflammation are associated with poor prognosis in lacunar infarctions. AB - Lacunar infarction has long been considered to be associated with good prognosis, however a significant percentage of these patients remain functionally dependent. In this study we sought to investigate the factors associated with poor outcome in patients with lacunar infarction. SUBJECTS AND METHODS: We have performed a secondary study in 113 patients with lacunar infarctions admitted within the first 24 h of symptom onset (mean age 70 years, 57.5% men). Blood pressure, body temperature, serum glucose levels, neurotransmitters and pro-inflammatory markers were measured at admission and during the first 72 h. Stroke severity was assessed by the Canadian Stroke Scale (CSS). Neuroimaging evaluation was performed at admission and between days 4 and 7. Poor functional outcome was considered as a Barthel index <85 at 3 months. RESULTS: 36 patients (31.9%) had poor outcome. Older age (p = 0.009), history of hypertension (p = 0.005), higher body temperature (p < 0.0001), systolic blood pressure (SBP) (p = 0.010), serum glucose (p = 0.002) and interleukin-6 (IL-6) (p < 0.0001) levels, as well as lower CSS score at admission (p < 0.0001) were all predictive factors of poor outcome in bivariate analyses. SBP at admission (OR 2.07, CI 95% 1.04-3.28, p = 0.015) was the only clinical predictor on multivariate analysis. When the logistic model was further adjusted for biomarkers of inflammation and excitotoxicity, IL-6 levels (OR 1.09, CI 95% 1.01-1.26, p = 0.003), but not SBP, was independently associated with poor outcome. This association persisted even after adjusting for potential predictors recorded during the first 72 h of hospitalization. CONCLUSION: High SBP and IL-6 levels on admission may predict poor outcome in patients with lacunar infarction. PMID- 16691021 TI - Subtypes and one-year survival of first-ever stroke in Chinese patients: The Nanjing Stroke Registry. AB - BACKGROUND: Morbidity and mortality of stroke have been investigated extensively in Western populations, while data concerning case fatality and cause of death after stroke are very limited in mainland China. This study aimed to analyze the 1-year survival and predictors of case fatality in Chinese patients with first ever stroke. METHODS: Subjects are patients registered in the Nanjing Stroke Registry Program. Information concerning cardiovascular risk factors and stroke characteristics were collected, and patients were followed after registration. Ischemic strokes were classified according to TOAST criteria as large-artery atherosclerosis (LAA), cardiac embolism stroke (CES), small-vessel stroke (SVS), or other determined and undetermined causes (UND). One-year case fatality was analyzed by the Kaplan-Meier method, and predictors of case fatality were evaluated by the Cox proportional hazards model. RESULTS: A total of 752 patients with first-ever stroke were included, of which 142 (18.9%) were identified as intracerebral hemorrhage (ICH), 120 (16.0%) as LAA, 123 (16.4%) as SVS, 160 (21.3%) as CES and 216 (28.7%) as UND. The overall survival rate was 86.4% at the end of the 1-year follow-up. Patients with SVS have the highest survival rate (92.7%), followed by UND (89.4%), CES (88.1%) and LAA (84.2%). Patients with ICH have the lowest survival rate (76.8%). Survival rates of patients with different subtypes of stroke presented a significant difference (chi2 = 19.3, p < 0.001). For patients deceased during the first year after the index stroke, 33.3% of deaths were caused by the first stroke, 18.6% by recurrent stroke, 16.7% by cardiovascular comorbidities, 14.7% by nonvascular conditions and 16.7% died of undetermined causes. Advanced age, hypertension, hyperlipidemia, diabetes mellitus (DM), atrial fibrillation (AF), history of transient ischemic attack and cigarette smoking were associated with an increased risk of death 1 year after stroke. CONCLUSIONS: The case fatality rate and predictors for mortality of Chinese patients with first-ever stroke are similar to those reported for other populations. The significant influence of cardiovascular disease on the first year survival rate emphasizes the importance of acute stroke management and control of hypertension, DM, AF and other predictors for decreasing case fatality and improving prognosis. PMID- 16691022 TI - A longitudinal study of health-related quality of life of first stroke survivors' close relatives. AB - BACKGROUND AND PURPOSE: Mental and social burdens of stroke patients' relatives have been investigated repeatedly, but there is little information about their quality of life (QoL). The purpose of the study was to describe the health related QoL of stroke patients' relatives at 3 points in time after the stroke. METHOD: Sixty-four closest relatives of patients suffering from a disabling stroke were questioned at admission to inpatient rehabilitation as well as 6 months and 1 year after discharge with the SF36 (short form 36-item questionnaire) and about probable covariates. RESULTS: During inpatient rehabilitation relatives' perception of their physical role as well as of all mental health scales was significantly lower than the normative data. We found improvements in all scales except physical functioning between inpatient rehabilitation and 6 months after discharge. Deteriorations between 6 months and 1 year occurred in physical functioning, role physical, social functioning, role emotional and mental health. CONCLUSIONS: Changes in perceived health-related QoL indicate that relatives go through different stages while coping with the chronic illness of their family member during rehabilitation and in the first year after discharge. These findings may aid support and counseling. PMID- 16691023 TI - ICAM-1 and VCAM-1 expression following aneurysmal subarachnoid hemorrhage and their possible role in the pathophysiology of subsequent ischemic deficits. AB - BACKGROUND: The pathophysiology of ischemic cerebral lesions following aneurysmal subarachnoid hemorrhage (SAH) is poorly understood. There is growing evidence that inflammatory reactions could be involved in the pathogenesis of such delayed occurring ischemic lesions. The aim of this study was to evaluate adhesion molecules with regard to these lesions following SAH. METHODS: Serum and cerebrospinal fluid (CSF) samples were taken daily from 15 patients up to day 9 after SAH and evaluated for intercellular adhesion molecule-1 (ICAM-1) and vascular adhesion molecule-1 (VCAM-1). RESULTS: CSF and serum samples correlated well during nearly the whole time course (p < 0.0001). A secondary increase in ICAM-1 and VCAM-1 in the serum and CSF correlated with an increase in flow velocity in the transcranial Doppler (p > 0.0001 and p < 0.007) but not to a delayed lesion in the CT scan. CONCLUSION: We believe that inflammatory processes are involved in the pathogenesis of cerebral vasospasm but they might only be a part of a multifactorial pathogenesis. PMID- 16691024 TI - Dissection of cervical arteries: Long-term follow-up study of 130 consecutive cases. AB - BACKGROUND AND PURPOSE: We describe the natural history, functional prognosis and long-term recurrences of patients with dissection of cervical arteries (DCA) in a sequential observational study. METHODS: We describe 130 patients with angiographically-proven DCA admitted to the Neurology Institute in Mexico City (Mexico), and analyzed clinical and neuroimaging data, treatment and outcome. Treatment with either anticoagulation or aspirin was decided by the primary physician. Primary outcome measures were recurrence (stroke and death) and clinical outcome at 6 months. Follow-up studies were performed to determine recanalization. RESULTS: Mean age was 35.4 years; 4 patients died (3%) and 126 were followed for 3,906 person/years; 17 patients (13%) had a heralding ischemic cerebral event (6 strokes, 11 TIAS) about 8 days before the diagnosis of DCA. After diagnosis, recurrent ischemic stroke occurred in 6 patients (4.8%) within the 2 first weeks (1.5 persons/1,000 follow-up years). No significant differences were found between aspirin and anticoagulation. Recanalization was more frequent in vertebral dissections. Complete recanalization of vertebral dissections was associated with a favorable prognosis [OR 3.2 (95% CI 1.1-8.8; p = 0.02)]. CONCLUSIONS: In Mexico, DCA affects young adults and may present with a heralding stroke or TIA. We found rare, early ischemic recurrences. Vertebral territory dissections had better prognosis than carotid ones, particularly in patients with demonstrated complete recanalization. PMID- 16691025 TI - Lobar brain hemorrhages and white matter changes: Clinical, radiological and laboratorial profiles. AB - BACKGROUND: White matter changes have several histopathologic correlates including cerebral amyloid angiopathy (CAA). The aim of this study was to characterize the clinical, laboratorial and neuroradiological profile of a CAA related lobar hemorrhages case series. METHODS: A cohort of 50 consecutive patients with cerebral lobar hemorrhages was studied and clinical, radiological data and ApoE polymorphisms were analyzed. White matter changes were graded and microbleeds were characterized according to number and location using T2* MRI. RESULTS: A statistically significant association was found between the prestroke cognitive performance and poststroke dementia and between hemorrhage volume and mortality. More severe white matter changes were found in probable CAA when comparing to possible CAA. The most prominent white matter lesions are associated with the presence and the number of microbleeds. The frequency of APOE epsilon 2 and epsilon 4 alleles was higher in this cohort when compared to a Northern Portuguese population. CONCLUSION: White matter changes are frequent in lobar hemorrhage patients and are associated with cortical microbleeds, the radiological hallmark of CAA. Therefore, white matter changes may be the sole phenotype of CAA and, potentially, involved in the pre-stroke cognitive impairment presented by the patients, which are genetically distinct from the population in general. PMID- 16691026 TI - Trojan horse meets magic bullet to spawn a novel, highly effective drug delivery model. AB - Because of their physicochemical properties and the selective receptor-mediated uptake of their core components, reconstituted high-density lipoproteins have unique advantages over conventional formulations to serve as targeted drug delivery agents. PMID- 16691027 TI - Nephrotoxicity due to combination antibiotic therapy with vancomycin and aminoglycosides in septic critically ill patients. AB - The aim of this prospective observational study was to evaluate the incidence of nephrotoxicity due to combination therapy with vancomycin and aminoglycosides in septic critically ill patients admitted to the intensive care unit. METHODS: Thirty consecutive critically ill patients were treated with vancomycin concurrent with aminoglycosides for sepsis. Inclusion criteria were: the need for mechanical ventilation and the presence of severe infection due to bacteria susceptible to vancomycin and aminoglycosides. Exclusion criteria were: age <18 years, impaired renal function (24-hour creatinine clearance <90 ml/min) or previous adverse reaction to either drug. Serum creatinine and urea concentrations, creatinine clearance, 24-hour urinary excretion of proteins, beta2-microglobulin and enzymes were measured immediately before starting therapy and at different times thereafter. RESULTS: Eleven of the 30 patients had a transient and modest increase in serum urea, 15 patients presented with urinary excretion of beta2-microglobulin and tubular enzymes, and 14 patients had urinary proteins. In the only patient with severe acute renal failure (serum creatinine 8.2 mg/dl), the clinical course was complicated by prolonged hypotension. CONCLUSION: Concurrent administration of vancomycin and aminoglycosides to critically ill septic patients with normal renal function at baseline induced mainly slight and transient toxic tubular effects. The only clinically significant nephrotoxic event occurred in a patient with septic shock. PMID- 16691028 TI - DNA-spermine and DNA-lipid aggregate formation visualized by fluorescence correlation spectroscopy. AB - BACKGROUND: Fluorescence correlation spectroscopy (FCS) can be used for the determination of diffusion coefficients of single molecules. Since diffusion coefficients are correlated with size and shape of the labeled species, FCS provides information on conformational changes in plasmids aggregates. METHODS: A 10-kbp plasmid stained with PicoGreen was condensed by spermine or liposomes formulated from cationic lipid and egg phosphatidylcholine. RESULTS: The diffusion coefficient of DNA increases from 1.0 x 10(-12) m2/s to 3.2 x 10(-12) m2/s by the addition of spermine, whereas the addition of cationic liposomes leads to complexes characterized by diffusion coefficients with values ranging from 1.7 to 1.9 x 10(-12) m2/s. CONCLUSIONS: FCS experiments allow determining the diffusion coefficients of DNA-containing aggregates which provide information regarding the topology and homogeneity of the aggregate. PMID- 16691029 TI - Efficacy and safety of fluvastatin-extended release in hypercholesterolemic patients: morning administration is equivalent to evening administration. AB - BACKGROUND: Flexibility in the time of administration of statin therapy is likely to improve patient compliance. This study compared the efficacy and tolerability of morning and evening administration of the extended-release formulation of fluvastatin (fluvastatin XL). METHODS: In this prospective, double-blind, multicenter, multiple dose study, 236 patients with type IIa/b hypercholesterolemia were randomized to receive fluvastatin XL, 80 mg, in the morning or evening for 8 weeks. RESULTS: At 8 weeks, low-density lipoprotein cholesterol levels were reduced by 34.5 and 35.0% in the morning and evening treatment groups, respectively (p = 0.0118 for non-inferiority of morning administration). There were no statistically significant differences between the morning and evening treatment groups in the changes in total cholesterol (p = 0.56), high-density lipoprotein cholesterol (p = 0.21), triglycerides (p = 0.13), apolipoprotein B (p = 0.66) and apolipoprotein AI (p = 0.88) at 8 weeks. The frequency of adverse events was slightly lower in the morning treatment group compared with the evening treatment group (27.4 vs. 35.5%). CONCLUSIONS: The efficacy and safety profiles of fluvastatin XL are equivalent for morning and evening administration. PMID- 16691030 TI - Modulation of atopy patch test and skin prick test by pretreatment with 1% pimecrolimus cream. AB - BACKGROUND: In a subgroup of patients with atopic eczema (AE), aeroallergens are relevant eliciting factors. The atopy patch test (APT) was proposed as inflammation model for AE. OBJECTIVE: It was the aim of this study to investigate the effect of pretreatment with 1% pimecrolimus cream (Elidel) on the APT and skin prick test (SPT). METHODS: In a randomized, controlled, double-blind study, 20 patients with AE and positive SPT and APT screening reaction to house dust mite Dermatophagoides pteronyssinus, cat dander, grass or birch pollen were enrolled (age 20 +/- 11 years, 55% males). For 2 weeks, patients twice daily applied pimecrolimus and vehicle control to marked fields on their backs and forearms. Then, APT was performed (200 index of reactivity/g extracts in petrolatum; Stallergenes, France) on both fields on the back and SPT was performed on the pretreated forearms. RESULTS: Including only patients with different readings (n = 13), stronger APT suppression of at least 1 ETFAD (European Task Force on Atopic Dermatitis) grade in the pimecrolimus area versus intraindividual control was observed in 10 of these patients after 48 and 72 h (p < 0.05; 90% CI 50.5-93.4). Including all 20 subjects, the analysis still showed a borderline significance compared with the vehicle (p = 0.0564). SPT with histamine and aeroallergens showed a median 7.5-10% reduction in actively pretreated areas (p = 0.086). Immunohistochemical analysis in 2 patients revealed an induction of interferon-gamma in primecrolimus-pretreated skin. CONCLUSION: APT can be used as a model for AE skin inflammation. It was shown for the first time that pimecrolimus pretreatment has a potential to suppress the development of lesions induced by aeroallergen exposure in patients with AE. PMID- 16691031 TI - Long-term immunologic effects of broad-spectrum aeroallergen immunotherapy. AB - BACKGROUND: Limited data exist regarding extended, long-term immunologic effects of immunotherapy in polysensitized individuals. To study possible long-term effects, skin tests and specific IgE levels were obtained from subjects who had previously received broad-spectrum aeroallergen immunotherapy years before. METHODS: Eighty-two subjects (78% male, mean age 23 years) previously enrolled in a randomized, placebo-controlled trial of immunotherapy for treatment of childhood allergic asthma were reevaluated in adulthood (mean follow-up interval, 10.8 years) by puncture skin tests and CAP-RAST levels for major aeroallergens. All completed at least 18 months (median 27 months) of maintenance active treatment or placebo injections without subsequent immunotherapy. RESULTS: At adult follow-up, 36% of all skin tests to treatment allergens among subjects who received immunotherapy (n = 41) had significantly reduced intensity versus 26% of skin tests among placebo recipients (n = 41; p = 0.03). No significant differences were noted for individual treatment allergens. No significant differences were observed in the long-term changes of serum-specific IgE antibody levels for all treatment allergens between immunotherapy treatment and placebo groups (p = 0.43). The treatment and placebo groups had a similar acquisition of new skin test sensitivities from time of randomization in the original childhood trial to debriefing (15 vs. 20%; p = 0.28) and to adult follow-up (30 vs. 31%; p = 0.75). CONCLUSIONS: Immunotherapy suppresses skin test sensitivity 8-16 years after discontinuation of treatment, but long-term effects on specific IgE levels in serum are not observed. Broad-spectrum immunotherapy does not appear to affect the acquisition of new inhalant sensitivities. PMID- 16691032 TI - Response of respiratory flour allergics in an ingested flour challenge may involve plasmacytoid dendritic cells, CD25+ and CD152+ T cells. AB - BACKGROUND: A number of occupational respiratory allergens are food related, and little is known about the responses these allergens elicit in sensitized persons that ingest them. METHODS: Nine respiratory flour-allergic volunteers were exposed in a double-blind placebo-controlled food challenge with flour. Responses were monitored by spirometry, acoustic rhinometry, determination of urinary methyl histamine and tryptase and flow cytometric evaluation of basophil, dendritic and T cell numbers and markers. RESULTS: Significant increases in serum tryptase (compared with placebo post-exposure levels) and methyl histamine and a coordinated decrease in blood basophils and nasal volume after ingestion of allergen compared with placebo suggest an allergic response to ingested allergen. There was no change in forced expiratory volume in 1 s. The number of blood plasmacytoid dendritic cells (DC), but not of myeloid DC, decreased after exposure (p = 0.001). DC HLA DR was reduced after both exposures (p < 0.001). Expression of CXCR4 on DC was reduced after allergen (p = 0.033) but not after placebo exposure. CD4+ T cell expression of CD25 was elevated after placebo (p = 0.021) but reduced after allergen provocation. The reduction in CD25 expression after allergen compared with placebo was significant (p = 0.024). CD152 was downregulated on these cells after allergen (p = 0.039) but less so after placebo exposure. CONCLUSION: Persons with respiratory allergy respond after ingestion of the relevant allergen. Response to this allergen challenge may selectively recruit plasmacytoid DC through CXCR4 and T cells expressing CD25 and CD152, which may be a regulatory phenotype. PMID- 16691033 TI - The pathogenesis of autosomal dominant polycystic kidney disease. AB - In individuals with autosomal dominant polycystic kidney disease (ADPKD), renal function deteriorates as the kidneys become replaced by multitudes of fluid filled cysts. Although the PKD genes were identified a decade ago, the pathway(s) leading from mutation to disease remain the subject of intense investigation. As a result of this work, it has become apparent that the polycystins are multifunctional proteins that, in the broadest sense, appear to be involved in the transduction of a number of environmental cues into appropriate cellular responses. It is likely that the central pathogenetic pathway for cystogenesis stems from de-differentiation of tubular epithelial cells. Available evidence indicates that loss of polycystin activity leads to subtle derangements of cell calcium regulation through several possible pathways. Abnormal cell calcium homeostasis might then lead to altered differentiation in affected cells. The study of the polycystins has revealed some entirely novel insights into fundamental cell biology but these have not yet been satisfactorily integrated into a verified pathogenetic pathway for the development of ADPKD. PMID- 16691034 TI - Renal vascular resistance in sepsis. AB - AIMS: To assess changes in renal vascular resistance (RVR) in human and experimental sepsis and to identify determinants of RVR. METHODS: We performed a systematic interrogation of two electronic reference libraries using specific search terms. Subjects were animals and patients involved in experimental and human studies of sepsis and septic acute renal failure, in which the RVR was assessed. We obtained all human and experimental articles reporting RVR during sepsis. We assessed the role of various factors that might influence the RVR during sepsis using statistical methods. RESULTS: We found no human studies, in which the renal blood flow (and, therefore, the RVR) was measured with suitably accurate direct methods. Of the 137 animal studies identified, 52 reported a decreased RVR, 16 studies reported no change in RVR, and 69 studies reported an increased RVR. Consciousness of animals, duration of measurement, method of induction of sepsis, and fluid administration had no effect on the RVR. On the other hand, on univariate analysis, size of the animals (p < 0.001), technique of measurement (p = 0.017), recovery after surgery (p = 0.004), and cardiac output (p < 0.001) influenced the RVR. Multivariate analysis, however, showed that only cardiac output (p = 0.028) and size of the animals (p = 0.031) remained independent determinants of the RVR. CONCLUSIONS: Changes in RVR during sepsis in humans are unknown. In experimental sepsis, several factors not directly related to sepsis per se appear to influence the RVR. A high cardiac output and the use of large animals predict a decreased RVR, while a decreased cardiac output and the use of small animals predict an increased RVR. PMID- 16691035 TI - Chronic inhibition of nitric oxide production aggravates diabetic nephropathy in Otsuka Long-Evans Tokushima Fatty rats. AB - BACKGROUND: Nitric oxide (NO) is known to play a role in diabetic nephropathy, but the molecular basis for this effect remains unclear. METHOD: Otsuka Long Evans Tokushima Fatty spontaneous diabetic rat models were used along with Long Evans Tokushima Otuska rat models as age-matched controls. Either L-arginine (a NO precursor) or L-NAME (a nitric oxide synthase inhibitor) was administered from the age of 22 weeks. Clinical parameters and serum and urinary NO2+NO3 levels were measured, in addition to renal histological findings and ED-1-positive cell counts in glomeruli. RESULTS: There were no significant differences in creatinine clearance between any of the groups at any point. The levels of urinary NO2+NO3 in the diabetic group were significantly lower than those in the control groups after 40 weeks; that in the L-NAME diabetic group was significantly lower than in the other diabetic groups at 52 weeks. Compared with the other diabetic groups, the L-NAME diabetic group had significantly higher urinary protein excretion levels, histological scores, and numbers of ED-1-positive cells in glomeruli. Diabetic rats administered L-arginine excreted more urinary protein than the diabetic controls. CONCLUSION: Diabetic nephropathy was exacerbated drastically by a nitric oxide synthase inhibitor and mildly by a NO precursor. These data suggested that NO may modify type 2 diabetic nephropathy in Otuska Long-Evans Tokushima Fatty rats through factors other than hemodynamics. PMID- 16691036 TI - FGF-23 and sFRP-4 in chronic kidney disease and post-renal transplantation. AB - BACKGROUND: The phosphatonins fibroblast growth factor-23 (FGF-23) and FRP-4 are inhibitors of tubular phosphate reabsorption that may play a role in the hyperphosphatemia associated with chronic kidney disease (CKD) or in the hypophosphatemia associated with renal transplants. METHODS: Plasma FGF-23, FRP 4, phosphorus and parathyroid hormone were measured in patients at all stages of CKD. Phosphate regulation of FGF-23 and secreted frizzled related protein-4 (sFRP 4) was examined in end-stage renal disease patients in the presence and absence of therapeutic phosphate binder usage. In renal transplant patients, plasma FGF 23, sFRP-4 and phosphorus concentrations were determined before and 4-5 days after transplantation. RESULTS: Plasma FGF-23 correlated with creatinine clearance (r2 = -0.584, p < 0.0001) and plasma phosphorus (r2 = 0.347, p < 0.001) in CKD patients and with plasma phosphorus (r2 = 0.448, p < 0.001) in end-stage renal disease patients. Phosphate binder withdrawal increased FGF-23 levels. In kidney transplant patients, dramatic decreases in FGF-23 (-88.8 +/- 5.4%) and phosphorus (-64 +/- 10.2%) were observed by 4-5 days post-transplantation. In patients with post-transplant hypophosphatemia, FGF-23 levels correlated inversely with plasma phosphorus (r2 = 0.661, p < 0.05). sFRP-4 levels did not change with creatinine clearance or hyperphosphatemia in CKD or end-stage renal disease patients, and no relation was noted between post-transplant sFRP-4 levels and hypophosphatemia. CONCLUSIONS: In CKD, FGF-23 levels rose with decreasing creatinine clearance rates and increasing plasma phosphorus levels, and rapidly decreased post-transplantation suggesting FGF-23 is cleared by the kidney. Residual FGF-23 may contribute to the hypophosphatemia in post-transplant patients. PMID- 16691037 TI - Decrease in salivary secretion by radiation mediated by nitric oxide and prostaglandins. AB - OBJECTIVE: In the present work, we evaluated the effect of exposing the submandibular glands (SMG) to radiation, studying different functional parameters such as salivary secretion, nitric oxide (NO) production, reactive oxygen species formation, prostaglandin (PGE) content and apoptosis. METHODS: We irradiated rats in the head and neck region with a single dose of gamma-ray radiation of 15 Gy. Two hours after radiation, we measured norepinephrine-induced salivary secretion. After that, the SMG were dissected, and in this tissue, we measured the activity of NO synthase (NOS), the PGE content, the amount of reactive oxygen species, apoptotic cells and mitochondrial inducible NOS (iNOS) expression. RESULTS: We found that radiation decreased salivary secretion when 10 and 30 microg/kg of norepinephrine was administered via the right femoral vein. We observed that iNOS activity was reduced and PGE content increased after radiation in SMG, indicating that NO and PGEs may participate in salivary secretion. The expression of mitochondrial NOS was increased after radiation leading to the formation of large amounts of NO that acts as a proapoptotic signal. In fact, we observed an augmentation in apoptotic cells. In this study, we also observed an increase in lipid peroxidation induced by radiation that may contribute to tissue damage. CONCLUSIONS: Our results indicate that radiation induced a decrease in salivary secretion and SMG iNOS activity, meanwhile the PGE content, the lipid peroxidation and apoptosis increased in the tissue. These modifications decrease salivary secretion. PMID- 16691038 TI - Thiopurine S-methyltransferase pharmacogenetics: genotype to phenotype correlation in the Slovenian population. AB - The toxicity of thiopurine drugs has been correlated to the activity of thiopurine S-methyltransferase (TPMT), whose interindividual variation is a consequence of genetic polymorphisms. We have herein investigated the relevance of some genetic markers for the prediction of thiopurine-related toxicities and to determine the genotype to phenotype correlation in the Slovenian population. The most prevalent mutant allele in the Slovenian population is TPMT*3A (4.1%), followed by TPMT*3C (0.5) and TPMT*3B (0.3), while the TPMT*2 allele was not found in any of the examined samples. TPMT enzyme activity distribution in the subgroup sample was bimodal and as such correlated with genetic data. Using a cutoff value of 9.82 pmol/10(7) RBC per h, the genetic data correctly predicted TPMT enzyme activity in 91.6% of the examined individuals. Pharmacogenetic TPMT analyses have therefore proved to have significant clinical implications for prediction of individuals' responses to treatment with thiopurine drugs in order to avoid possible life-threatening therapy-related toxicities. PMID- 16691039 TI - [Analysis of the nursing interventions performed by public health nurses in health centers using the NIC]. AB - PURPOSE: The purpose of this study was to identify nursing interventions performed by public health nurses in health centers. METHOD: Data was collected by the taxonomy of Nursing Intervention Classification (NIC 3rd: 486 nursing interventions) from 131 public health nurses in health centers and analyzed using descriptive statistics. RESULT: As its result, more than 50% of public health nurses performed 137 nursing interventions at least monthly. The most frequently used intervention class was "activity and exercise management", followed by "physical comfort promotion", "community health promotion", "life span care", "coping assistance", "self care facilitation", "information management", "nutrition support", "community risk management" and "patient education". One hundred twenty nursing interventions were rarely performed by 90% or more of the nurses. Most of them were in the physical complex domain. CONCLUSION: In conclusion, 137 interventions were performed by public health nurses at least monthly. NIC is helpful to build a standardized language for public health nursing. PMID- 16691040 TI - [A comparison of interventions recorded in nursing notes between acute and subacute stage after a cerebrovascular accident]. AB - PURPOSE: This study aimed to: 1) determine the core nursing interventions, and 2) compare acute interventions with subacute interventions recorded in the nursing notes of patients with cerebrovascular accidents (CVA). METHODS: The nursing records covering the first 10 days of 30 patients with a CVA who were admitted from January to December 2004 at C University Hospital in Korea were examined. Data was collected using the nursing interventions classification (NIC) from January to April 2005. Finally, data analysis was carried out using mean, SD, and paired t-test according to domains, classes, and interventions. RESULTS: The most frequent nursing intervention at both stage was "Neurologic monitoring". There were differences in interventions belonging to the "Physiological: complex," "Behavioral," "Safety," and "Health system" domains between the acute and subacute stages. The frequency of interventions belonging to the "Immobility management," "Neurological management," "Tissue perfusion management," "Patient education," "Risk management," "Health system mediation," and "Information management" classes at the acute stage was higher compared to the subacute stage. CONCLUSIONS: This study found out that nurses relatively recorded more nursing interventions during the acute stage hence the unsuccessful documentation of the subacute stage particularly in describing the specific nursing interventions at this stage. PMID- 16691041 TI - [The effects of inhalation of essential oils on the body weight, food efficiency rate and serum leptin of growing SD rats]. AB - PURPOSES: This experimental study was designed to verify the effect of inhalation of essential oils on body weight, feed intake, food efficiency rate and serum leptin. METHODS: The subjects of this study were 90 growing SD rats (46 males and 44 females). They were allocated into one of four groups, the Fennel group, Patchouli group, Bergamot group and control group. The experimental treatment was the inhalation of aromatherapy essential oils which was applied two times a day for 10 minutes each during 8 weeks. To evaluate the effects, body weight, feed intake, food efficiency rate and serum leptin were measured before and after the treatment. The collected data was analyzed by repeated measures of Kolmogorov smirnov test and Normal Q-Q plot for normality, Kruskal Wallis test and chi2-test for experimental effects with the SPSS program. RESULTS: The food efficiency rate was significantly lower in the Patchouli group and Fennel group than in the Bergamot group and control group (P=.000). No significant group effects were found for SD rat's body weight, feeding amount and serum leptin. CONCLUSION: In conclusion, these findings indicate that the inhalation of essential oils could be effective in lowering the food efficiency rate rather than the feed intake. PMID- 16691042 TI - [Impact of clinical nurses' power and empowerment on job satisfaction and organizational commitment]. AB - PURPOSE: This study was performed to analyze effects of the power and empowerment on job satisfaction and organizational commitment. This study was based on the Kanter's theory of organizational empowerment. METHOD: A predictive, non experimental design was used to test the model in a sample of 688 nurses working in 7 university hospitals that have over 500 beds in Seoul, Kyunggi and Kangwon provinces. The data were collected from December, 2003 to January, 2004. It was analyzed with descriptive statistics and Pearson correlation of SPSS and with path analysis of LISREL. RESULT: The formal and informal power had direct effects on empowerment. Formal power also had direct effects on informal power. Empowerment had direct effects on job satisfaction and organizational commitment. Organizational commitment had direct effects on job satisfaction. There was positive effects in all of the variables. CONCLUSION: The positive changes show on personal behaviors and attitudes when the nurses who have formal and informal power are empowered. These findings would be important resource to nurse administrators for clinical implication. PMID- 16691043 TI - [Yakson vs. GHT therapy effects on growth and physical response of preterm infants and on maternal attachment]. AB - PURPOSE: This study is aimed to confirm the effects of Yakson therapy on the growth and physical response of preterm infants, and maternal attachment to them compared with GHT therapy. METHOD: The design of this study is nonequivalent control group with repeated measuring by quasi experimental study. The subjects are preterm infants in 26 - 34 gestational age hospitalized in the NICU of 4 university hospitals with an experimental group of 15 and a control group of 14. Yakson therapy consists of three phases: laying a hand, caressing by hand, and laying a hand again taking 5 minutes for each phase. RESULT: As a result of administering Yakson therapy to preterm infants; the average weight gain of the Yakson group was higher than that of the GHT group, but there is no significant difference between groups. The oxygen saturation and maternal attachment difference between the Yakson and the GHT group were not significant. Significant differences in the average daily increase of oral intake and apical pulse rate were observed between the Yakson group and GHT group. CONCLUSION: These data suggested that Yakson therapy may be an effective nursing intervention which can facilitate growth and physical response of preterm infants. PMID- 16691044 TI - [A comparative study on family perception between abused children and normal children by Kinetic Family Drawing]. AB - PURPOSE: The study attempted to find family perception differences between abused children and normal children by Kinetic Family Drawing. METHOD: The subjects of the study consisted of two groups, 143 abused who were in the upper 25th percentile, and 150 normal who were in the lower 25th percentile. Collected Kinetic Family Drawings were divided into five dimensions such as actions, human figure characteristics, dynamics, styles and symbols, and they was analyzed with SPSS/WIN 10.0. RESULTS: In the perception about their family in action dimension, their family in figure characteristics dimension, their family in dynamics dimensions, and their family in symbols dimension, there is a sharp contrast between the two groups. CONCLUSION: Putting these results together, abused children feel lower self-esteem and feel more sense of alienation in their family than normal children do. In addition, abused children perceive their parents as negative and aggressive people. PMID- 16691045 TI - [The effect of Tai Chi movement in patients with rheumatoid arthritis]. AB - PURPOSE: This study was performed to verify the effect of Tai Chi exercise on patients with rheumatoid arthritis particularly their level of pain, fatigue, sense of balance and daily life performance (ADL). METHOD: It employed a non equivalent control group pre- and post-test design. The research instruments used in this study were pain, fatigue, sense of balance and ADL. Thirty-two patients in the experimental group carried out 50 minutes of Tai Chi exercise for 12 weeks, and 29 patients in the control group did not. Before and after the experiment, both groups were tested for pain, fatigue, sense of balance and ADL. Collected data were processed using the SPSS/WIN 10.0 program analyzed by the frequency, percentage, chi2-test, and t-test. RESULTS: Pain and fatigue significantly decreased in the experimental group. However the improvement in ADL of the rheumatoid arthritis patients was not statistically significant but their sense of balance was enhanced significantly. CONCLUSION: Tai Chi exercise is an effective nursing intervention that can be used for rheumatoid arthritis patients. PMID- 16691046 TI - [Development and application of an online alternative therapy and health promotion class]. AB - PURPOSE: This study was to develop an online "alternative therapy and health promotion" class for university students and to evaluate its changes. METHOD: The online class was developed based on the Instructional Systems Development (ISD) model and model of Web-Based Instruction (WBI) developmental process. This was a quasi- experimental, one group pretest-posttest design. The subjects of this study were 130 students in 3 universities, and they were provided the cyber class for 16 weeks. Data was analyzed by descriptive and plural answer statistics, and paired t-test. RESULTS: The cyber class was developed in five steps: analysis, design, data collection and reconstruction, programming and publishing, and evaluation. The results of program evaluation were positive, which included learning 3.47, system 3.57, and learning satisfaction 3.64 on the scale of 5. The posttest scores of cognition and reliability of alternative therapy were higher than pretest scores. The posttest scores of health promoting lifestyle (t=-5.051, p=.000) and perceived health status (t=2.979, p=.003) were significantly higher than those of the pretest. CONCLUSION: These results suggest that the cyber class is a positive method in increasing a cognition, reliability of alternative therapy, and is effective to improve a health promotion lifestyle and perceived health status for the university students. PMID- 16691047 TI - [A paradigm analysis related to spiritual experiences focused on Christian of patients with terminal cancer]. AB - PURPOSE: The main purpose of this study was to develop a substantive theory on the process of the spiritual experience in Christian terminal cancer patients in the context of Korean society and culture. The question for the study was "What is the spiritual process in Christian terminal cancer patients?" METHOD: The research method used was the Grounded Theory Method developed by Strauss and Corbin (1998). Participants for this study in total were 9 Christian terminal cancer patients. Data was collected using in-depth interviews during April 2003 to March. 2004. Data collection and analysis were carried out at the same time. RESULT: From the analysis 58 concepts and 20 categories emerged. The categories were presented into a paradigm, which consisted of condition-actions/interactions consequences. The theoretical scheme was described by organizing categories. In total, 4 stages were developed from the condition-actions/interactions consequences. Throughout these stages, the "overcoming process of unbalanced interconnectedness" was the core category discovered. CONCLUSION: This study provides a framework for the development of individualized care interventions in the "overcoming process of unbalanced interconnectedness" for Christian terminal cancer patients. PMID- 16691048 TI - [The effects of a rehabilitation program on physical health, physiological indicator and quality of life in breast cancer mastectomy patients]. AB - PURPOSE: The purpose of this study was to investigate the effects of a rehabilitation program on physical health, physiological indicators and quality of life in breast cancer mastectomy patients. METHODS: The subjects included thirty-one patients with breast cancer (17 in the experimental group and 14 in the control group). The subjects in the experimental group participated in a rehabilitation program for 10 weeks, which was composed of an exercise program, teaching, counseling and support for 2 sessions per week. RESULTS: There was a significant increase in flexion, internal rotation and external rotation but no significant increase in extension in the experimental group compared to the control group. The total cholesterol, triglyceride, HDL, LDL, and CD56 in the experimental group compared to the control group was not significantly decreased after the rehabilitation program. Compared to the control group, quality of life in the experimental group was significantly improved and fatigue in that group was significantly decreased after the rehabilitation program. CONCLUSION: The 10 week rehabilitation program showed a large affirmative effect on physical health, physiological indicators and quality of life in breast cancer mastectomy patients. PMID- 16691049 TI - [The effect of music therapy on anxiety and depression in patients undergoing hemodialysis]. AB - PURPOSE: The purpose of this study was to determine the effects of music therapy on anxiety and depression in patients undergoing hemodialysis. METHOD: The study was designed using a nonequivalent control group pretest-posttest design. The subjects consisted of 36 patients (Experimental group: 18, CONTROL GROUP: 18) who received hemodialysis in three hospitals located in Seoul. The measures were a Music Preference Questionnaire (MPQ), anxiety measurement, and depression measurement. Data was collected from December 26, 2004 to April 2, 2005 through questionnaires. The collected data was analyzed by the SPSS 10.0 program. RESULT: The first hypothesis that patients undergoing hemodialysis who received music therapy would have less anxiety than patients undergoing hemodialysis who did not receive music therapy was supported (F=8.05, p=.008). The second hypothesis that patient undergoing hemodialysis who received music therapy would have less depression than patients undergoing hemodialysis who did not receive music therapy was supported (F=11.86, p=.002). CONCLUSION: The results of this study suggest that music therapy may be applied as a method of nursing intervention contributing to the improvement of quality of life by reducing their anxiety and depression of patients undergoing hemodialysis. PMID- 16691050 TI - [The factors influencing Korean health behavior]. AB - PURPOSE: This study was performed to investigate the factors influencing health behavior. METHOD: Data used in this study was drawn from a 2001 National Health survey done by Korea Institute for Health and Social Affairs. Number of samples were 5,085 people between age 20 years and 69 years in whole country. In this study, we modified Green and Kreuter's Precede-Proceed model to analyse influence of factors in health behavior. Hierarchical regression were used in the analysis. RESULT: 1) Factors that had statistically significant positive relation with overall health behavior were age, educational level, income level, disease in the family, medical examination, subjective weight, and concern about health, 2) Factors that had statistically significant negative relation with overall health behavior were sex, subjective health, stress, and degree of physical activity. CONCLUSION: 1) It would be necessary to set the target group for the health promotion in advance. 2) It was very important to adopt easy and efficient methods to change the health behavior of target group, for example, ability to control stress. PMID- 16691051 TI - [The development & effect of a tailored fall prevention exercise for older adults]. AB - PURPOSE: This study was conducted to develop and to determine the effect of an tailored falls prevention exercise for older adults. METHOD: Subjects consisted of 59 older adults (experimental group: 29, control group: 30) living at nursing homes. Experimental group participated in tailored falls prevention exercise for 16 weeks (3 times a week, 50 min every session). Data were collected before the exercise, 16 weeks & 24 weeks after the beginning of exercise. RESULTS: 1) the experimental group significantly improved the muscle strength of hip extensor & flexor, knee extensor & flexor, ankle dorsiflexor, & plantar flexor compared to the control group. 2) the experimental group significantly decreased the mean time for 10 times chair stand. 3) the experimental group significantly increased the standing time on one leg and the number of heel raise for 30 seconds compared to the control group. In addition more older adults in the experimental group completed the tandem stance and semi-tandem stance for 10 seconds than the control group. 4) The experimental group significantly decreased the mean time of 6m walk and the fall frequency compared to the control group. CONCLUSIONS: This results suggest that tailored falls prevention exercise for older adults can improve muscle strength, static & dynamic balance and decrease the fall frequency of older adults. PMID- 16691052 TI - [Effects of backrest position on central venous pressure and intracranial pressure in brain surgery patients]. AB - PURPOSE: This study was done to investigate the effects of backrest elevation of 0 degree and 30 degrees that minimize the risk of increasing ICP when CVP is measured. METHODS: Subjects were sixty-four patients who stayed in the neurosurgical intensive care unit after brain surgery at two university-based hospitals. CVP, blood pressure, heart rate and ICP were measured along with position changes in order of backrest position with primary 30 degrees backrest position, 0 degree backrest position and secondary 30 degrees backrest position. For data analysis, one-group, repeated-measures analysis of variance design was used in SAS program. RESULTS: Backrest elevations from 0 degree to 30 degrees did not alter the CVP without increasing the ICP. Therefore, 30 degrees backrest position is a preventive position without increasing ICP. CONCLUSION: 30 degrees backrest position might be appropriate for brain injury patients when CVP is measured. PMID- 16691053 TI - [The substitution effect of leadership substitutes for transformational leadership in nursing organization]. AB - PURPOSE: This paper was conducted to examine the effects of transformational leadership behaviors, within the substitutes for leadership model (Kerr & Jermier, 1978). METHOD: Data was collected from 181 staff nurses in 3 general hospitals, with self-reporting questionnaires (MLQ developed by Bass, rd-SLS developed by Podsakoff, et al., and MSQ developed by Weiss, et al.). Descriptive statistics, factor analysis, Cronbach's alpha and moderated regression analysis were used. RESULT: 1) The transformational leader behaviors and substitutes for leadership each had correlations with job satisfaction. 2) The total amount of variance accounted for by the substitutes for leadership was substantially greater than by the transformational leadership behaviors. 3) Few of the substitutes variables moderated the relationships between the transformational leader behaviors and job satisfaction in a manner consistent with that specified by Howell, Dorfman, and Kerr (1986). CONCLUSION: The finding of this study suggest that leaders need to have a better understanding of those contextual variables that influence job satisfaction. Thus future research should focus attention on the moderating effects of substitutes, as well as the things that leaders can do to influence them. In addition, it may be good to examine the effects of substitutes on other criterion variables. PMID- 16691054 TI - [A study on the related factors of the wellbeing of family caregivers in elderly with stroke]. AB - PURPOSE: The purpose of this study was to identify the factors related to the wellbeing of the family caregivers of the elderly with a stroke. METHODS: The subjects of this study were 199 elderly treated in four oriental hospitals in Korea, and their primary family caregivers. The data was collected by interviews and a self-reported questionnaire, during the period from October, 2003 to April, 2004. RESULTS: The results of this study were as follows. The mean score of wellbeing of family caregivers was 60.6412.63. The factors related to wellbeing of family caregivers were sex, age, education, depression, illness severity, ADL, paralysis, and speech disability in elderly characteristics. Among family caregivers characteristics, education, relation, and burden were significantly related. In situational variables, family income and the previous relationship between the elderly and family caregivers were related to wellbeing. Stepwise multiple regression analysis revealed that the most powerful predictor of wellbeing was the burden of family caregivers. A combination of the depression of elderly and age of family caregivers accounted for 50.3% of the variance of wellbeing. CONCLUSIONS: On developing the nursing intervention for improving wellbeing of family caregivers, many factors should be considered, especially caregiver burden, and elderly depression. PMID- 16691055 TI - [A nationwide survey on current conditions of school health education]. AB - PURPOSE: The purpose of this study was to analyze and assess the current situation of Korea's school health education program and to establish measures to efficiently carry out school health education in Korea. METHOD: The survey was conducted through the internet with the health educators of elementary, middle, and high schools nationwide to assess the current condition of school health education programs, and 2,459 samples were collected which accounted for 23.4% of the total respondents. RESULTS: According to school health educators on the enforcement of health education, the higher the education became, the less the health education was enforced. The enforcement rate was 96.9% in elementary schools, 76.7% in middle schools, and 67.3% in high schools. The major reasons were found as difficulty in securing class time (54.5%) and other excessive workloads (20.9%). As a result of the health education awareness survey, over 99% answered that health education is needed, over 80% answered that the education requires independent health textbooks, and over 95% answered that health educators are suitable for the person in charge of the education. CONCLUSION: This study will be a useful in establishing a detailed policy on enhancing school health education in the future. PMID- 16691056 TI - [Development of objectives in nursing clinical education based on the Nursing Core Competencies]. AB - PURPOSE: The purpose of this study was to set up a Nursing Core Competencies required for staff nurses and to set up Objectives for Nursing Clinical Education based on the Nursing Core Competencies. The objectives in this study are to be achieved ultimately through clinical practice because it is a common avenue of work and the basic objective regardless of the education system and curriculum. METHOD: A nursing Core Competencies were established by literature review and verified by 15 experts. Nursing Clinical Education Objectives were established by literature review and analysis, and a survey for validity using a five point Likert scale was given to 257 nursing professors, 503 head-nurses, 509 staff nurses who had less than 3 years clinical experience in 34 general hospitals and 738 senior student nurses from 81 nursing colleges. RESULT: Nine nursing core competencies were set up. In addition 39 Objectives for each of the nursing clinical core competencies were set up. CONCLUSION: In conclusion, this study will contribute to professional nursing education to provide comprehensive nursing care by applying knowledge to nursing practice to achieve the Nursing Core Competency as a professional nurse. PMID- 16691057 TI - [Effects of constraint-induced movement using self-efficacy enhancing strategies on the upper extremity function of chronic hemiplegic patients]. AB - PURPOSE: The purpose of this study was to investigate the effects of constraint induced (CI) movement using self-efficacy on U/E function of chronic hemiplegic patients. CI movement discourages the use of the unaffected U/E, combined with intensive training of the affected U/E. METHOD: A non-equivalent pretest-posttest design was used. Study subjects were 40 hemiplegic patients selected from 2 different community health centers. The experimental subjects participated in the CI movement program for 6 hours daily over a period of two weeks. The exercises for affected U/E consisted of warming up, main exercise and ADL practice. To encourage the participants' behaviors self-efficacy enhancing strategies were used, which included performance accomplishment, vicarious experience, verbal persuasion and emotional arousal. RESULT: After 2 weeks of treatment, the grip power, pinch power, wrist flexion/extension, elbow flexion, and shoulder flexion/extension were significantly higher in the experimental subjects than in the control subjects. However, there was no significant difference in hand functions of the two groups. CONCLUSION: The above results show that the constraint-induced movement using self-efficacy could be an effective nursing intervention for improving U/E function of chronic hemiplegic patients. Long term studies are needed to determine the lasting effects of constraint-induced movement. PMID- 16691058 TI - [Analysis of smoking and smoking cessation related nursing research in Korea and its future direction]. AB - PURPOSE: The purpose of this study was to analyze the contents and trend of smoking and smoking cessation related nursing research in Korea and to suggest the directions for future research. METHOD: Eight nursing professional Journals published by the Korean Academic Nursing Society and by 7--adult, community, psychiatric, fundamental, obstetrics, pediatrics, administrative--nursing societies from beginning edition to October 2005 were selected. They were examined for the proportion of published research, participants, research designs, key words, measurement tools, and the intervention outcomes in the case of the experimental research. RESULT: Of the research analyzed, 87 studies were smoking and smoking cessation related research. The Journals, which had published the most number of smoking and smoking cessation research were J of Korean Academic Society, J of Korean Adult Nursing Society, J of Korean Community Nursing Society. The most popular research design was an descriptive design. The major concepts studied were related psychosocial variables. Among 87 studies, only 11 were experimental research. CONCLUSION: Research on smoking and smoking cessation in the nursing discipline are increasing. However more research to test the effectiveness of nursing intervention programs are needed. PMID- 16691059 TI - Testing a CCR5 drug? Avoid mosquito bites. PMID- 16691060 TI - The changing spectrum of pulmonary disease in patients with HIV infection on antiretroviral therapy. PMID- 16691061 TI - Safety and acceptability of cellulose sulfate as a vaginal microbicide in HIV infected women. AB - OBJECTIVES: Few studies of topical microbicides have assessed their safety in HIV infected women. We conducted this study to evaluate the safety and acceptability of 6% cellulose sulfate (CS) gel as a vaginal microbicide in sexually abstinent and active HIV-infected women. METHODS: Fifty-nine HIV-infected women were enrolled in a randomized double-blind placebo-controlled study comparing 6% CS to placebo gel used for 14 days. Sexually abstinent women applied gel once or twice daily and sexually active women used gel once daily. RESULTS: CS gel was safe with no reported severe or life-threatening adverse events (AE). Thirty-nine (66%) of the participants experienced urogenital AE judged as probably or possibly related to gel. The majority (51%) of these participants reported only mild events. Fewer women (62%) who used CS experienced urogenital AE than those assigned to placebo gel (70%) (P = 0.59). Eleven (19%) women experienced intermenstrual bleeding judged to be probably or possibly related to gel use (four in the CS and seven in the placebo gel group). There was no increase in AE by frequency of gel use or sexual activity with the exception of abdominal/pelvic pain which was noted more frequently with twice daily use among sexually abstinent women. Women and men found the gel highly acceptable. CONCLUSIONS: This Phase I study demonstrated that CS vaginal gel was safe, well tolerated and acceptable by HIV-infected women and their male partners. Thus, further development of CS is warranted as a potential method to prevent HIV transmission and acquisition. PMID- 16691062 TI - Impact of baseline viral load and adherence on survival of HIV-infected adults with baseline CD4 cell counts > or = 200 cells/microl. AB - BACKGROUND: Baseline plasma HIV RNA levels > 100 000 copies/ml have been associated with elevated mortality rates after the initiation of HAART. There is uncertainty regarding the optimal strategy for patients with high plasma HIV RNA but CD4 cell count > or = 200 cells/microl. OBJECTIVE: To evaluate the impact of baseline plasma HIV RNA on survival among patients with CD4 cell counts > or = 200 cells/microl. METHODS: Patients were stratified by plasma HIV RNA, CD4 cell count and adherence level. Mortality rates were evaluated using Kaplan-Meier methods and Cox regression. RESULTS: Among 1166 patients initiating HAART with a CD4 cell count > or = 200 cells/microl, a baseline HIV RNA > or = 100 000 copies/ml was statistically associated with elevated mortality among non-adherent patients (log-rank P = 0.032), but not for adherent patients (log-rank P = 0.690). In a multivariate Cox model comparing patients with a baseline CD4 cell count > or = 200 cells/microl and a baseline plasma HIV RNA < 100 000 copies/ml, the mortality rate was statistically similar among patients with a baseline CD4 cell count > or = 200 cells/microl and a baseline plasma HIV RNA > or = 100 000 copies/ml (relative hazard, 1.21; 95% confidence interval, 0.89-1.65; P = 0.232). CONCLUSION: HIV RNA > or = 100 000 copies/ml was only associated with mortality among HIV-infected patients initiating HAART with CD4 cell counts > or= 200 cells/microl if the patients were non-adherent. PMID- 16691063 TI - Therapy of "SHIV" infected macaques with liposomes delivering antisense interleukin-4 DNA. AB - BACKGROUND/OBJECTIVES: To explore the effects of antisense (AS) interleukin (IL) 4 on virus replication and CD8+ T-cell responses in lymph nodes and blood of macaques infected with simian human immunodeficiency virus, SHIV(89.6)P. METHODS: Six macaques were inoculated with simian human immunodeficiency virus (SHIV(89.6)P). Seven days later, four of the animals were given 1 mg AS IL-4 plasmid complexed with Megafectin liposome, intravenously, and two of these received a second injection of the same material on day 9. All six macaques were killed at 2 weeks post infection (pi) and monitored for viral RNA and CD8+ T cells in blood and lymph nodes by real-time reverse transcriptase-polymerase chain reaction, flow cytometry and immunohistochemistry. RESULTS: In contrast to the lymph nodes from virus control animals, the lymph nodes of AS IL-4-treated animals had a significant reduction in viral loads and reduced depletion of cells from the nodes. There was an increase in CD8+ T cells in the nodes, and many of the cells expressed granzyme B, suggesting functional activation. This trend of virus reduction and increased CD8+ T cell numbers was also reflected in blood. CONCLUSIONS: The therapeutic effect of the AS IL-4 suggests indirectly that the acute immunosuppressive disease caused by SHIVs is mediated, in part, by IL-4 that causes enhanced virus replication by suppressing anti-viral CD8+ T-cell responses, and that this effect was reduced by treatment of the animals with AS IL-4. PMID- 16691064 TI - Atazanavir and lopinavir/ritonavir: pharmacokinetics, safety and efficacy of a promising double-boosted protease inhibitor regimen. AB - OBJECTIVE: To assess the pharmacokinetics and tolerability of lopinavir (LPV), ritonavir (RTV) and atazanavir (ATV) as a double-boosted protease inhibitor regimen in HIV-infected adults. METHODS: Sixteen patients who started LPV/RTV (400/100 mg b.i.d.) and ATV (300 mg q.d.) were enrolled in the study group (arm A). LPV pharmacokinetics were compared to those of two historical groups: arm B, 15 patients who received LPV/RTV (400/100 mg b.i.d.); and arm C, 25 patients who received LPV/RTV/saquinavir (SQV) (400/100/1000 mg b.i.d.). ATV pharmacokinetics were compared to those of 15 consecutive patients who received ATV and RTV (300/100 mg q.d.) (arm D). Drug concentrations were measured by HPLC. RESULTS: LPV concentrations were significantly higher in arm A than in arms B and C. Median (interquartile range) LPV area under the curve (AUC)0-12 values were 115.7 (99.8-136.5), 85.2 (68.3-109.2) and 85.1 (60.6-110.1) microg/h/ml, respectively. C(max) values were 12.2 (10.7-14.5), 9.5 (6.8-13.9) and 10.0 (6.9-13.6) microg/ml, respectively. C(min) values were 9.1 (7.1-10.4), 5.6 (4.7-8.2) and 5.5 (4.2-7.5) microg/ml, respectively. No difference was observed for ATV AUC0-24 or C(max) between arms A and D. ATV C(min) values were 1.07 (0.61-1.79) in arm A and 0.58 (0.32-0.83) in arm D (P = 0.001). Treatment was not discontinued in any patient because of adverse effects. At 24 weeks, viral load was < 50 copies/ml in 13 of 16 patients. CONCLUSIONS: The combination of ATV and LPV/RTV provided high plasma concentrations of both PI, which seemed to be appropriate for patients with multiple prior therapeutic failures, yielding good tolerability and substantial antiviral efficacy. PMID- 16691065 TI - Relationship between antiretrovirals used as part of a cART regimen and CD4 cell count increases in patients with suppressed viremia. AB - BACKGROUND: It is unknown if the CD4 cell count response differs according to antiretroviral drugs used in combination antiretroviral therapy (cART) in patients with maximal virological suppression [viral load (VL) < 50 copies/ml]. OBJECTIVES: To compare the change in CD4 cell count over consecutive measurements with VL < 50 copies/ml at both time-points according to nucleoside backbones and other antiretrovirals used. METHODS: Generalized linear models, accounting for multiple measurements within patients, were used to compare CD4 cell count changes after adjustment for antiretrovirals, time from starting cART, age, CD4 at first VL < 50 copies/ml, prior antiretroviral treatment, and change in CD4 cell count since starting cART. RESULTS: We studied 28418 instances of VL < 50 copies/ml in 4041 patients. The mean annual change in CD4 cell count was +45.5/microl [95% confidence interval (CI) +39.4 to +51.6/microl). Comparing two drug nucleoside backbones, there was a lower annual change in CD4 cell count for zidovudine/lamivudine (n = 13038; -15.4/microl; P = 0.012) and for those on tenofovir (n = 1809; -27.3/microl; P = 0.029) compared to lamivudine/stavudine (n = 7339). Compared to the boosted-protease inhibitor regimen (n = 5915), use of an abacavir-based triple-nucleoside regimen was associated with a lower annual change in CD4 cell count (n = 2504 pairs; -26.1/microl; P = 0.011). CONCLUSIONS: A nucleoside backbone of zidovudine/lamivudine or any tenofovir-based backbone was associated with significantly poorer increases in CD4 cell count compared to a nucleoside backbone of stavudine/lamivudine, as was an abacavir-based triple nucleoside regimen compared to a boosted protease inhibitor regimen. Long-term studies are needed to determine whether the differences in immunological response seen here translate into differences in the risk of clinical disease. PMID- 16691066 TI - A trial of SGN-00101 (HspE7) to treat high-grade anal intraepithelial neoplasia in HIV-positive individuals. AB - OBJECTIVES: To test a therapeutic vaccine consisting of a fusion of the human papillomavirus (HPV) 16 E7 protein and the Mycobacterium bovis heat shock protein 65 (SGN-00101) to treat high-grade anal intraepithelial neoplasia (HG-AIN) in HIV positive individuals. DESIGN: A phase I/II trial with three cohorts of five participants each, sequentially assigned to receive 100, 500 or 1000 microg SGN 00101, injected three times subcutaneously in alternating thighs at 4-week intervals. Anal disease was assessed at baseline, 8, 12, 24 and 48 weeks and was classified as the more severe of anal cytology and anal biopsy. Anal HPV DNA was detected using L1 consensus primer-based PCR followed by type-specific probing and dot-blot hybridization (DBH). HPV16, 18 and 31 DNA copy numbers were measured using quantitative real-time PCR. SETTING: University-based research clinic. PARTICIPANTS: Thirteen HIV-positive men and two HIV-positive women with HG-AIN. RESULTS: There were no drug-related serious adverse events or significant changes in HIV viral load and CD4/CD8 ratio. At 48 weeks, two of five participants in both the 100 and 500 microg cohorts regressed to AIN 1 and one of five participants in the 1000 microg cohort regressed to atypical squamous cells of undetermined significance (ASC-US). All participants had at least one oncogenic HPV type at baseline. Three of five (60%) participants who regressed to AIN 1 or ASC-US became HPV-negative using DBH and real-time PCR, compared with none of 10 participants with no clinical response (P = 0.02). CONCLUSIONS: SGN-00101 was well tolerated in HIV-positive individuals, with preliminary evidence for clinical activity. PMID- 16691067 TI - Efficacy of early treatment of acute hepatitis C infection with pegylated interferon and ribavirin in HIV-infected patients. AB - BACKGROUND: Treatment of acute hepatitis C (HCV) in HIV-infected patients has been poorly addressed. OBJECTIVE: To evaluate the efficacy and tolerability of a 24 week course of pegylated interferon alfa 2a (PegIFNalpha2a) and ribavirin for the treatment of acute HCV infection in HIV-infected patients. METHODS: This was a prospective pilot study of 25 consecutive HIV-infected men with acute HCV infection defined by documented HCV seroconversion to anti-HCV positive antibody and positive qualitative HCV RNA measurement. Patients with detectable HCV RNA (> 50 IU/ml) 12 weeks after diagnosis were offered treatment with PegIFNalpha2a (180 microg/week) and ribavirin (800 mg/day) for 24 weeks. Sustained virological response was defined by a negative qualitative HCV RNA measurement 24 weeks after the end of treatment. RESULTS: At baseline, 23 patients were taking HAART, 23 patients had HIV RNA < 200 copies/ml and a median CD4 count of 345 cells/microl. Only one patient, with genotype 3 HCV, had a spontaneous clearance of HCV RNA. Of the remaining 24 patients, four refused anti-HCV therapy, ribavirin was contraindicated in one and 19 initiated anti-HCV therapy. Median time between acute HCV diagnosis and initiation of study treatment was 14 weeks. Of the 14 patients who have achieved the post-treatment follow-up at 24 weeks, 10 had a sustained virological response (71%). Study treatment was well tolerated, with no change in CD4 cell count. CONCLUSION: Early treatment of acute HCV infection with PegIFNalpha2a and ribavirin for 24 weeks yields a high sustained virological response rate in HIV-infected patients. PMID- 16691068 TI - Generic fixed-dose combination antiretroviral treatment in resource-poor settings: multicentric observational cohort. AB - BACKGROUND: The use fixed-dose combination (FDC) is a critical tool in improving HAART. Studies on the effectiveness of combined lamivudine, stavudine and nevirapine (3TC/d4T/NVP) are scarce. OBJECTIVE: To analyse 6861 patients in a large observational cohort from 21 Medecins Sans Frontieres (MSF) HIV/AIDS programmes taking 3TC/d4T/NVP, with subcohort analyses of patients at 12 and 18 months of treatment. METHODS: Survival was analysed using Kaplan-Meier method and factors associated with progression to death with Cox proportional hazard ratio. RESULTS: Median baseline CD4 cell count at initiating of FDC was 89 cells/microl [interquartile range (IQR), 33-158]. The median follow-up time was 4.1 months (IQR, 1.9-7.3). The incidence rate of death during follow-up was 14.2/100 person years [95% confidence interval (CI), 13.8-14.5]. Estimates of survival (excluding those lost to follow-up) were 0.93 (95% CI, 92-94) at 6 months (n = 2,231) and 0.90 (95% CI, 89-91) at 12 months (n = 472). Using a Cox model, the following factors were associated with death: male gender, symptomatic infection, body mass index < 18 kg/m and CD4 cell count 15-50 cells/microl or < 15 cells/microl. Subcohort analysis of 655 patients after 1 year of follow-up (M12 FDC cohort) revealed that 77% remained on HAART, 91% of these still on the FDC regimen; 5% discontinued the FDC because of drug intolerance. At 18 months, 77% of the patients remained on HAART. CONCLUSIONS: Positive outcomes for d4T/3TC/NVP are reported for up to 18 months in terms of efficacy and safety. PMID- 16691069 TI - Effect of hepatitis C infection on progression of HIV disease and early response to initial antiretroviral therapy. AB - OBJECTIVES: To describe the effect of hepatitis C virus (HCV) on the progression of HIV disease and on early changes in the CD4 cell count and HIV viral load after HAART initiation. DESIGN AND METHODS: Data were from a longitudinal medical records review project conducted in over 100 US medical clinics from 1998 to 2004. We analysed data from HIV-infected patients who received antiretroviral therapy (ART), calculated adjusted hazard ratios describing the hazard of death or progression to an AIDS-defining opportunistic illness (AIDS-OI) associated with prevalent HCV infection, and estimated the change in CD4 cell count and HIV viral load after HAART initiation, stratified by HCV status. RESULTS: A total of 10 481 HIV-infected individuals were followed for a median of 1.9 years; 19% had HCV. HCV infection was not associated with progression to AIDS-OI or death after controlling for important confounding conditions. Factors significantly confounding the risk of both death and diagnosis of an AIDS-OI were alcoholism, drug-induced hepatitis, and the type of ART prescribed. Acute and chronic hepatitis B infection confounded the risk of AIDS-OI diagnosis. During the 12 months after starting HAART, proportional increases in CD4 cell counts did not differ between HCV-infected and HCV-uninfected individuals. Likewise, the short term change in viral load did not differ. CONCLUSION: In our cohort, HCV did not increase the risk of death or AIDS-OI, and did not affect the early immunological or virological response to initial HAART. Clinicians should evaluate patients with HCV for other, manageable problems, including alcoholism and other viral hepatitis. PMID- 16691070 TI - Mortality and causes of death in adults receiving highly active antiretroviral therapy in Senegal: a 7-year cohort study. AB - OBJECTIVES: To evaluate survival and investigate causes of death among HIV-1 infected adults receiving HAART in Senegal. DESIGN: An observational prospective cohort. METHODS: Mortality was assessed in the first patients enrolled between August 1998 and April 2002 in the Senegalese antiretroviral drug access initiative. First-line regimen combined two nucleoside reverse transcriptase inhibitors and either a non-nucleoside reverse transcriptase inhibitor or a protease inhibitor. The most likely causes of death were ascertained through medical records or post-mortem interviews (verbal autopsy). RESULTS: Four hundred and four patients (54.7% women) were enrolled in the study and were followed for a median of 46 months (interquartile range: 32-57 months) after HAART initiation. At baseline, 5% were antiretroviral therapy (ART) non-naive, 39 and 55% were respectively at CDC stage B and C, median age, CD4 cell count and viral load were 37 years, 128 cells/microl and 5.2 log cp/ml, respectively. Ninety-three patients died during follow-up and the overall incidence rate of death was 6.3/100 person years [95% confidence interval (CI), 5.2-7.7]. During the first year after HAART initiation, 47 patients died and seven were lost to follow-up, yielding to a probability of dying of 11.7% (95% CI, 8.9-15.3%). The death rate, which was highest during the first year after HAART initiation, decreased with time yielding a cumulative probability of dying of 17.4% (95% CI, 13.9-21.5%) and 24.6% (95% CI, 20.4-29.4%) at 2 and 5 years. Causes of death were ascertained in 76 deaths. Mycobacterial infections, neurotropic infections and septicaemia were the most frequent likely causes of death. CONCLUSIONS: This study underlines the early mortality pattern after HAART initiation and highlights the leading role of mycobacterial infections in the causes of death. PMID- 16691071 TI - Alcohol use before sex and HIV acquisition: a longitudinal study in Rakai, Uganda. AB - OBJECTIVE: To examine the association between alcohol use and HIV acquisition. DESIGN AND METHODS: We examined alcohol use before sex and incident HIV in a population-based cohort in Rakai, Uganda, between 1994 and 2002. Adjusted incidence rate ratios (adjIRR) of HIV acquisition and 95% confidence intervals (CI) were estimated by Poisson multivariate regression. We also estimated adjusted prevalence rate ratios to assess the association between alcohol use and the number of sex partners and consistency of condom use. RESULTS: In 6791 men and 8084 women HIV incidence was 1.4 per 100 person-years and 1.5 per 100 person years, respectively. After adjustment for sociodemographic and behavioral factors, the risks of HIV when one partner consumed alcohol before sex were: adjIRR 1.67, 95% CI 1.17-2.40 among men, and adjIRR 1.40, 95% CI 1.02-1.92 among women, and when both partners consumed alcohol the risks were adjIRR 1.58, 95% CI 1.13-2.21 among men, and adjIRR 1.81, 95% CI 1.34-2.45 among women. Alcohol use was significantly associated with inconsistent condom use and multiple sexual partners in both sexes. CONCLUSION: The use of alcohol before sex increases HIV acquisition. A reduction of alcohol use should be incorporated into HIV prevention programmes. PMID- 16691072 TI - Baseline HIV RNA and the when to start question: time to stop asking this question? PMID- 16691073 TI - Immunosuppression and GB virus C-RNA detection among HIV-infected patients in Cambodia. AB - In this survey, 213 patients in an antiretroviral treatment programme in Phnom Penh, Cambodia, were tested for GB virus C (GBV-C) RNA before treatment initiation. Most had advanced HIV infection, only 34 having CD4 cell counts > 200 cells/microl. GBV-C-RNA was detected in 35 patients. The proportion with positive GBV-C-RNA decreased dramatically with CD4 cell counts < 100 cells/microl. In multivariate analysis, low CD4 cell counts, tuberculosis, anaemia, and traditional medicine were independently and negatively associated with GBV-C-RNA detection. PMID- 16691074 TI - High prevalence of human papillomavirus infection in the anus, penis and mouth in HIV-positive men. AB - Human papillomavirus (HPV) types are associated with squamous cell cancers. HIV infection is linked with a higher prevalence of anal HPV infection. It is important to assess whether HPV is present in other body parts involved in sexual practices to establish a cancer prevention program. A high prevalence of high risk HPV types was present in the anus, penis and mouth (78, 36 and 30%, respectively) in a cohort of HIV-infected males (men who have sex with men and heterosexual), without evidence of pathology in these areas. PMID- 16691075 TI - "Serosorting" in casual anal sex of HIV-negative gay men is noteworthy and is increasing in Sydney, Australia. AB - This paper addresses the question of whether HIV-negative gay men engage in "serosorting" in casual encounters. Serosorting, defined as engaging in unprotected anal intercourse with casual partners who they report to be HIV negative, has been increasing among HIV-negative gay men in Sydney. Prevention and intervention programmes are urgently needed to alert HIV-negative gay men to the risks associated with "serosorting", and remind them of the need for consistent condom use. PMID- 16691076 TI - Skin rash associated with atazanavir: report of three cases. PMID- 16691077 TI - Research design determines what can be known about modes of HIV transmission. PMID- 16691078 TI - Unexpected improvement of sexual dysfunction during atazanavir therapy. PMID- 16691080 TI - Dramatic interaction between levothyroxine and lopinavir/ritonavir in a HIV infected patient. PMID- 16691081 TI - Haemolytic anaemia associated with efavirenz. PMID- 16691082 TI - Leishmania-HIV co-infection: an emerging problem in India. PMID- 16691083 TI - HIV optimism does not explain increases in high-risk sexual behaviour among gay men of positive or negative HIV status in Sydney, Australia. PMID- 16691084 TI - Musculoskeletal pain and menopausal status. AB - OBJECTIVES: The authors examined whether self-reported menopausal status is associated with musculoskeletal pain in a multiethnic population of community dwelling middle-aged women after considering sociodemographics, medical factors, smoking, depression, and body mass index using a cross-sectional study design. METHODS: Participants were 2218 women from the Study of Women's Health Across the Nation assessed at the time of their third annual follow-up exam. Two dependent variables were derived from a factor analysis of survey questions about pain. These 2 outcomes were Aches and Pains, derived from 5 of 6 pain symptom questions and Consultation for Low Back Pain, derived from 1 question. RESULTS: Prevalence of aches and pains was high, with 1 in 6 women reporting daily symptoms. Compared with premenopausal women, those who were early perimenopausal (P=0.002), late perimenopausal (P=0.002), or postmenopausal (P<0.0001) reported significantly more aches and pains in age-adjusted analysis. With complete risk factor adjustment, postmenopausal women still reported significantly greater pain symptoms (P=0.03) than did premenopausal women. Menopausal status was marginally related to consulting a healthcare provider for back pain. DISCUSSION: This study demonstrates an association between pain and self-reported menopausal status, with postmenopausal women experiencing greater pain symptoms than premenopausal women. PMID- 16691085 TI - The cognitive risk profile for pain: development of a self-report inventory for identifying beliefs and attitudes that interfere with pain management. AB - OBJECTIVES: An extensive body of research suggests that maladaptive beliefs about chronic pain can have a negative impact on patient adherence and treatment response. A series of studies to develop and validate a clinically-based, self report instrument for pain beliefs, the Cognitive Risk Profile for Pain (CRPP), was undertaken. We sought to expand the existing body of knowledge for pain beliefs by development of an instrument with a somewhat different content and format than prior pain belief measures, and a primary focus on clinical risk assessment for treatment planning. METHODS: Test development and evaluation procedures were applied in the initial stages of CRPP development. We report here on a series of studies to evaluate and refine the structure and content of the CRPP, and to establish its internal reliability, concurrent, and criterion validities. A 68-item version of the CRPP was evaluated, including a total risk score and 9 scale scores: philosophic beliefs about pain (PB), denial that mood affects pain (MP), denial that pain affects mood (PM), perception of blame (BL), inadequate support (IS), disability entitlement (DE), desire for medical breakthrough (MB), skepticism of multidisciplinary approach (SM), and conviction of hopelessness (CH). The CRPP was administered to two large samples of chronic pain outpatients (n=499; 125) in conjunction with other self-report scales for pain and associated beliefs, behaviors, and psychopathology. In a final study, treatment outcome measures were obtained for a subsample of chronic pain patients (n=91) to evaluate criterion validity. RESULTS: Confirmatory factor analyses showed improved fit for the CRPP scale structure after elimination of 15 items. The resulting 53-item CRPP was found to have good internal consistency for the full score (alpha=0.82) and 7 of 9 scales, with moderate consistency for scales BL and MB. Low to moderate scale intercorrelations were found. Correlations with pain and psychosocial measures suggested good construct validity for the majority of individual scales and total score. Results were inconsistent for scale MP. Multivariate analyses of variances (MANOVAs) based on tertile split of total risk scores showed significant main effects across pain, mood, productivity, and sleep ratings at 3 and 6-month treatment follow-ups. Analyses of clinically significant treatment changes (ie, 2 points on a 11-point Numerical Rating Scales) showed significantly higher prevalence of treatment "failures" at 6 months among CRPP high-risk patients, but no significant differences at 3 months. DISCUSSION: Results provide initial support for the CRPP as a reliable, valid, and useful measure of general cognitive risk for pain management. Results were supportive of the content and reliabilities of the majority of scale scores. Scales for denial of mood impact on pain, perception of blame, and desire for medical breakthrough will require further evaluation. Data indicate an association of CRPP total risk with multidimensional outcome from medical treatment of chronic pain, supporting relevance to treatment planning. The unique content and format of the CRPP may be useful in some clinical pain settings. Possible applications of the CRPP for risk assessment and treatment planning for chronic pain are discussed. PMID- 16691086 TI - Factors that influence the applicability of sham needle in acupuncture trials: two randomized, single-blind, crossover trials with acupuncture-experienced subjects. AB - OBJECTIVES: In recent years the retractable type of sham needle has been anticipated to be a possible solution for masking patients in acupuncture research. However, this needle has been intended mainly for acupuncture-naive subjects. The authors' goal in this study was to assess the validity of the retractable type of sham needle. METHODS: The authors conducted two randomized, single-blind, crossover trials with acupuncture-experienced subjects. Different acupuncture points were used in each trial (a LI-4 trial and a BL-23 trial). The subjects received two sessions of different stimulations in each trial. A Park Sham Needle was used in one session, a genuine acupuncture needle in the other. RESULTS: In the LI-4 trial, all of the 21 subjects (100%) felt penetration with the genuine needle, but only 7 of the 20 subjects (35%) felt a similar sensation with the sham needle (P=0.0002). Fifteen of the 21 subjects (71%) felt a dull sensation with the genuine needle, but only 4 of the 20 subjects (20%) felt a similar sensation with the sham needle (P=0.01). In the BL-23 trial, 14 of the 20 subjects (70%) felt penetration with the genuine needle and 10 of the 20 subjects (50%) felt "penetration" with the sham needle (P=0.39). Eight of the 20 subjects (40%) felt a dull sensation with the genuine needle and 2 of the 20 subjects (10%) did with the sham needle (P=0.109). CONCLUSIONS: Potential factors that influence the applicability of "placebo" needling include not only inter-tester variability but also the patient's knowledge and experience of acupuncture, acupuncture point selection, the visual impact of needling, and so on. PMID- 16691087 TI - Use of telemedicine in chronic pain consultation: a pilot study. AB - OBJECTIVES: Telemedicine has been used extensively in various settings, including monitoring patient treatment response and counseling. However, there are few data on the application of telemedicine to chronic pain patients. The present study was the first pilot project to determine whether telemedicine technology for chronic pain consultation was feasible, cost-saving, and satisfactory to patients and pain physicians. METHODS: A prospective pilot study was conducted on chronic pain patients requiring follow-up consultations using telemedicine technology. Patients were interviewed by phone following the consultation. RESULTS: Eleven telemedicine anesthesia consultations involving eight patients (age 42+/-9 years; six men, two women) were performed. All were follow-up consultations. The average distance from patients' home to the clinic was 314+/-170 km. The reasons for consultation were for update of patient progress (10/11), medication change (6/11), and counseling (3/11). The time to complete the consultation was 24.5+/ 9.5 minutes. The data for the time and the cost that the patient spent on the consultation are presented as median and 25% to 75% interquartile range. Patients having telemedicine consultations spent 0.9 hours (0.83-1) and Canadian dollar 3 (dollar 2-4) versus an estimate of 8 hours (6-8) and Canadian dollar 80 (dollar 46-260) for a conventional consultation (both P<0.005). Telemedicine consultation was found to be highly satisfactory to the patient and the consulting and attending anesthesiologists. CONCLUSIONS: This pilot study indicates that telemedicine follow-up consultations for chronic pain patients are feasible and cost-saving. Patients and anesthesiologists were highly satisfied with telemedicine consultation. Patients reported a significant saving in time and cost compared with a conventional consultation. PMID- 16691088 TI - Incidence of phantom phenomena including phantom limb pain 6 months after major lower limb amputation in patients with peripheral vascular disease. AB - OBJECTIVES: Contentions exist regarding the true incidence of phantom limb pain (PLP) and other associated post-amputation phenomena. Recognizing and understanding these phenomena would assist in the rehabilitation of amputees. This study was designed to investigate all post-amputation phenomena in a homogenous group of amputees. METHODS: Prospective amputees were recruited prior to amputation of a lower limb due to peripheral vascular disease. All survivors were followed 6 months after surgery and interviewed to identify post-amputation phenomena, including phantom sensations, PLP, and stump pain. RESULTS: Sixty amputees were recruited and 52 survived until the 6-month interview. Phantom sensations were universal, and aspects of the non-painful phenomena, including kinetic, kinesthetic, and exteroceptive components, were identified at varying rates within the sample. PLP was found in 78.8% of the survivors, and 51.2% had stump pain. Super-added phenomena occurred in 15.4%. Links were found between PLP and stump pain (P=0.01) and PLP and the ability to move the phantom (P=0.01). No link was found between PLP and telescoping of the phantom (P=0.47). CONCLUSIONS: Phantom phenomena are associated with many myths. This study starts to unravel myth from fact, but further study is required before this enigmatic condition and its influence on rehabilitation are fully understood. PMID- 16691089 TI - Evaluation of allodynia and pain associated with postherpetic neuralgia using current perception threshold testing. AB - OBJECTIVES: Postherpetic neuralgia has various clinical features, and the implicated pathophysiologic mechanisms are controversial. This study was carried out to clarify the roles of peripheral sensory nerves in the production of allodynia and ongoing pain. Current perception threshold (CPT) testing was used to evaluate the sensory function. METHODS: The intensities of ongoing pain and dynamic allodynia were assessed using a numeric rating scale (0-10). Assessment of sensory nerve function was performed by a series of 2,000-, 250-, and 5-Hz stimuli using CPT testing. These measurements were made in ipsilateral and contralateral area. RESULTS: CPTs at all frequencies in the ipsilateral area were significantly higher than those in the contralateral area. There were significant and inverse correlations between the intensity of allodynia and CPTs at all frequencies. No correlation was found between the intensity of ongoing pain and CPTs at any frequency. There was no correlation between the intensity of ongoing pain and the intensity of dynamic allodynia. CONCLUSIONS: The intensity of dynamic allodynia in postherpetic neuralgia correlates with the preserved functions of Abeta, Adelta, and C fibers. In contrast, the intensity of ongoing pain does not correlate with either the preserved function of C fibers or the intensity of dynamic allodynia. Therefore, it is suggested that postherpetic neuralgia might be a pain syndrome including both peripheral and central mechanisms. PMID- 16691090 TI - Treatment of chronic low back pain with successive injections of botulinum toxin a over 6 months: a prospective trial of 60 patients. AB - OBJECTIVES: The aim of this study was to evaluate the effects of two successive neurotoxin treatments for chronic low back pain using multiple pain rating scales in an open-label, prospective study. METHODS: Adult patients with chronic low back pain received multiple paraspinal muscle injections with a maximum dosing of 500 units of botulinum A toxin per session. Those with a beneficial clinical response received a second treatment at 4 months. Pain was assessed by visual analog scale (VAS), modified low back pain questionnaire (OLBPQ), and a clinical low back pain questionnaire (CLBPQ) at baseline, 3 weeks, 2 months, 4 months, and 6 months after the first treatment. RESULTS: Eighteen women and 42 men, ages 21 to 79 years (mean 46.6 years), with low back pain of a mean duration of 9.1 years were included. Significant improvement in back and radicular pain occurred at 3 weeks in 60% and at 2 months in 58% of the cohort. Beneficial clinical response to the first injection predicted response to reinjection in 94%. A significant minority of patients had a sustained beneficial effect from the first injection at 4 (16.6%) and 6 months (8.3%). Two patients had a transient flu-like reaction after the initial treatment. CONCLUSIONS: Botulinum toxin A improves refractory chronic low back pain with a low incidence of side effects. The beneficial clinical response is sustained with a second treatment. PMID- 16691091 TI - Manual therapy, physical therapy, or continued care by the general practitioner for patients with neck pain: long-term results from a pragmatic randomized clinical trial. AB - OBJECTIVES: The authors' goals were to compare the effectiveness of manual therapy (MT; mainly spinal mobilization), physical therapy (PT; mainly exercise therapy), and continued care by the general practitioner (GP; analgesics, counseling and education) over a period of 1 year. METHODS: One hundred eighty three patients suffering for at least 2 weeks from nonspecific neck pain were randomized to receive a 6-week treatment strategy of MT once a week, PT twice a week, or GP care once every 2 weeks. The primary outcome measures were perceived recovery, severity of physical dysfunctioning, pain intensity, and functional disability. RESULTS: The differences between groups considered over 1 year were statistically significant (repeated measurements analyses P<0.001 to P=0.02) for all outcomes but borderline for the Neck Disability Index (P=0.06). Higher improvement scores were observed for MT for all outcomes, followed by PT and GP care. The success rate, based on perceived recovery after 13 weeks, was 72% for MT, which was significantly higher than the success rate for continued GP care (42%, P=0.001) but not significantly higher compared with PT treatment (59%, P=0.16). The difference between PT and GP approached statistical significance (P=0.06). After 1 year the success rates were 75%, 63%, and 56%, respectively, and no longer significantly different. CONCLUSIONS: Short-term results (at 7 weeks) have shown that MT speeded recovery compared with GP care and, to a lesser extent, also compared with PT. In the long-term, GP treatment and PT caught up with MT, and differences between the three treatment groups decreased and lost statistical significance at the 13-week and 52-week follow-up. PMID- 16691092 TI - The attitudes to back pain scale in musculoskeletal practitioners (ABS-mp): the development and testing of a new questionnaire. AB - OBJECTIVES: Little is known about practitioners' beliefs and attitudes to the treatment of low back pain, and whether these influence their clinical decisions, intervention strategies, and patient-centered outcomes. This study aimed to develop, test, and explore the underlying dimensions of a new questionnaire, the Attitudes to Back Pain Scale (ABS), in a specific group of clinicians, practitioners who specialize in musculoskeletal therapy. METHODS: Items for the draft questionnaire were derived from interviews with practitioners (chiropractors, osteopaths, and physiotherapists). The draft questionnaire (52 items) sought to assess practitioners' attitudes concerning role and self-image plus their beliefs about treatment goals and prognosis of low back pain. The questionnaire was sent to a random selection of 300 practitioners from each professional group, and 546 (61%) responded. Split-sample analyses were performed using exploratory and confirmatory factor analysis. RESULTS: Separate exploratory analyses were done for attitudes concerned with personal interaction (34 items) and attitudes about treatment orientation (18 items), producing six domains: limitations on sessions, psychologic, connection to health care system, confidence and concern, reactivation, and biomedical. Confirmatory analyses indicated that the model tested presented a good fit. Validity interviews revealed high agreement of categorization and low levels of difficulty in categorizing the items. CONCLUSIONS: The internal structure of the new questionnaire not only shows excellent psychometric properties and good face validity, but also has the added advantage of being developed with a specific clinical context in mind. Additional evaluation is required to fully describe the psychometric integrity of this instrument. PMID- 16691093 TI - Mothers' responses to children's pain. AB - OBJECTIVES: This study was aimed at identifying mothers' responses to children's pain, evaluating whether these could be organized into different types of responses, and developing a questionnaire to assess these responses. METHODS: Common responses to children's pain were identified on the basis of a review of the literature and interviews with mothers of pediatric patients with pain. Categories reflecting these parenting behaviors were generated for a questionnaire on Adult Responses to Children's Symptoms, which was administered to 145 mothers of pediatric patients aged between 8-18 years referred for medical evaluation of abdominal pain. RESULTS: Factor analysis using principal components extraction with oblique rotation yielded 3 factors. Factor 1, Protect, reflected caretaking behaviors that placed the child in a passive sick role. Factor 2, Minimize, reflected criticism of the child's pain behavior. Factor 3, Encourage and Monitor, reflected encouragement of the child's activity while monitoring the child's symptoms. Subscales based on these correlated factors had good internal consistency. DISCUSSION: Results suggest that mothers' responses to children's pain behavior may be classified into 3 distinct categories. Additional research is needed to assess whether observational methodologies would yield a similar typology of parents' responses to children's pain. Psychometric properties of the Adult Responses to Children's Symptoms should be examined in larger samples and in studies of the relation of the subscales to related constructs (eg, measures of parenting beliefs and behavior) and to children's pain behavior. PMID- 16691094 TI - Impact of opioid use on outcomes of functional restoration. AB - OBJECTIVES: To examine the effect of opioid use on psychological function, physical functioning, and return-to-work outcomes of a multidisciplinary rehabilitation program (MRP) for chronic pain. METHODS: The participants were 127 patients with on-the-job injuries who had completed an MRP between 2001 and 2003. Opioid use was controlled by the patients' treating physicians (who were not affiliated with the MRP) and was assessed via patient self-report at the time of admission to the program and discharge. Other measures included pretreatment and posttreatment assessments of depression, pain severity, perceived disability, and physical ability (floor-to-waist lifting capacity). Return-to-work outcomes were obtained via follow-up phone calls approximately 6 months posttreatment. RESULTS: Significant improvements from pretreatment to posttreatment were evidenced on all psychological and physical measures for both opioid users and nonusers. Further, there were no significant posttreatment differences between opioid and nonopioid users on psychological, physical, or return-to-work outcomes. DISCUSSION: The role of opioids in the treatment of chronic pain continues to be controversial. Despite a lack of definitive data on their effectiveness, opioids continue to be prescribed, and thus patients using opioids continue to present for multidisciplinary rehabilitation. Although further exploration is warranted, results of the current study suggest that opioid use during rehabilitation does not necessarily preclude treatment success. PMID- 16691095 TI - Complex regional pain syndromes in children and adolescents: regional and systemic signs and symptoms and hemodynamic response to tilt table testing. AB - OBJECTIVE: Complex regional pain syndromes (CRPS) involve neuropathic limb pain and localized circulatory abnormalities. The authors hypothesized that (1) pediatric CRPS patients exhibit systemic autonomic symptoms and orthostatic and/or cardiac sympatho-vagal dysregulation and (2) their orthostatic regulation differs from healthy controls and pediatric patients with postural orthostatic tachycardia syndrome (POTS). METHODS: CRPS children and adolescents (n=20) underwent a 6-week trial of physical therapy and cognitive-behavioral treatment. Measures included pain and function scores, regional and systemic autonomic symptom profiles, heart rate and blood pressure with tilt, heart rate variability indices, and baroreflex gain. Systemic autonomic symptoms were recorded in 55 healthy pediatric controls. Tilt responses in CRPS patients were compared with those of 21 POTS patients and 39 healthy controls. RESULTS: CRPS patients' regional autonomic symptoms, pain, and limb function improved over 6 weeks (P<0.01). At baseline CRPS patients reported more systemic autonomic symptoms than controls (P<0.05). Tilt table test showed orthostatic stability, but the mean heart rate increase with tilt was greater in CRPS patients than controls (P<0.001). POTS patients showed significant increases with tilt in mean heart rate and diastolic and systolic blood pressures compared with controls (P<0.001). There were significant increases in the mean systolic and diastolic blood pressures in POTS compared with CRPS patients but no difference in the mean heart rate between groups. DISCUSSION: CRPS patients reported multiple regional and systemic autonomic symptoms that improved during the study course, and they experienced minimal and transient tilt table-induced hemodynamic changes compared with POTS patients but relatively similar to controls. PMID- 16691096 TI - Caveats to a dosing survey of chronic pain patients. PMID- 16691097 TI - Lumbar radiofrequency neurotomy. PMID- 16691098 TI - Radiofrequency facet denervation: a randomized control placebo versus sham procedure. PMID- 16691099 TI - New nuclear options--the AP1000. PMID- 16691100 TI - The contemporary materials cycle for radioactive 137Cs in the United States. AB - The materials cycle for (137)Cs, a low-level radioactive material of interest from a security perspective (as a possible source for "dirty bombs") as well as for its extensive industrial and medical uses, has been characterized for the United States for the year 2000. The focus is on products utilizing the isotope rather than on isotope production and subsequent disposal as a result of nuclear power generation. The results indicate that, during 2000, of the 1.5 PBq of (137)Cs that entered use, 94% was contained in sources in imported devices; the amounts in domestic source material recycling (4%) or as imported source materials (2%) were trivial by comparison. Losses from use were about 0.5 PBq; of this amount 86% was by radioactive decay, 11% was active source material that was recovered and recycled, and 3% was source material sent to low-level disposal sites. The current stock of (137)Cs in use is about 20 PBq; this stock is currently growing by more than 1 PBq per year (the difference between inputs to and losses from use). As a result, the security challenge related to monitoring stock in use is increasing by around 5% per year. PMID- 16691101 TI - Internal dose assessment of natural uranium from drinking water based on biokinetic modeling and individual bioassay monitoring: a study of a Finnish family. AB - Since the later 1960's, a nationwide survey on natural radionuclides in drinking water showed high concentrations of natural uranium (U) in Finland, especially in uraniferous granite areas. In order to assess the radiation dose from the natural uranium to individuals, the concentrations of natural uranium in drinking water of the drilled wells were determined by radiochemical and alpha spectrometric methods. Uranium contents were measured in the urinary samples of five members of a Finnish family by means of inductively coupled plasma-mass spectrometry. Correspondingly, theoretical biokinetic modeling of natural uranium incorporated for the same persons were performed with the aid of follow-up interviews. The ICRP biokinetic compartmental model and the age-dependent transfer rates for uranium were used to model the intake, transfer, distribution, retention, and excretion of (234)U and (238)U, respectively, from the drinking water for each person of the family. The organ absorbed dose, equivalent dose, and effective dose were evaluated for each family member at time intervals using specific effective energy values calculated by the SEECAL program and compared with recommended values. The modeled urinary excretion rates were found to be mostly higher than the measured values by a factor of three. The mean annual effective dose for this family is 8 muSv y(-1). By comparing the measured and calculated data, estimation of retrospective radiation exposure based on biokinetic modeling and bioassay method is enhanced and, vice versa, the biokinetic and dosimetric models are tested and verified. PMID- 16691102 TI - Bone marrow dose estimates from work-related medical x-ray examinations given between 1943 and 1966 for personnel from five U.S. nuclear facilities. AB - Inclusion of dose from work-related medical x-ray examinations with occupational external dose in an epidemiological study may reduce misclassification of exposures and provide more accurate assessment of leukemia risk from occupational exposure to ionizing radiation. In a multi-site leukemia case-control study, annual bone marrow doses due to work-related x-ray examinations given between 1943 and 1966 were estimated for cases and controls employed at five nuclear facilities. Only active bone marrow dose from photofluorographic chest and routine lumbar spine x rays were included. Bone marrow dose assigned for a single exposure ranged from 1.0 to 1.4 mGy. Mean and median cumulative bone marrow doses for each of the five sites from work-related x-ray examinations ranged from 2.0 to 14 mGy and 2.1 to 8.8 mGy, respectively. Results suggest that bone marrow dose from work-related photofluorographic and lumbar spine x-ray examinations given during the time period of this study may be significant compared to occupational bone marrow dose. PMID- 16691103 TI - Indications of the neutron effect contribution in the solid cancer data of the A bomb survivors. AB - Risk estimates for radiation-induced cancer are primarily based on the follow-up of the Japanese A-bomb survivors. Their exposures were due to gamma rays and neutrons, and, currently--with the assumed low RBE = 10 of neutrons and reference to the colon dose--the late radiation effects are almost fully attributed to the gamma rays. Solid cancer risk estimates for different organ sites are assessed here, and an inconspicuous but statistically highly significant trend of larger values is found for the organs closer to the body surface; i.e., the organs with less body shielding and, therefore, with larger neutron dose-fractions. Underestimation of the RBE of neutrons can explain this apparent correlation. The trend of ERR/Gy vs. depth ceases to be statistically significant for RBE values close to 100. The suggestion of high RBE values and the corresponding reduction of gamma-ray risk estimates is found to be in line with log-likelihood computations in terms of AMFIT, which provide for the solid cancer mortality of the A-bomb survivors the minimum deviance for RBE = 100 with a 95% confidence lower limit of 25. The present assessment had to use the data made publicly available by RERF. In this form they contain city-, sex-, age-, and dose categories, but--instead of a separate neutron-dose category--only the mean neutron dose for each data cell. The tentative conclusions that are here obtained should, therefore, be examined by a more definitive analysis, either in terms of grouped data with a separate classification of neutron doses or, ideally, in terms of person by person calculations to be performed at RERF with individually estimated neutron doses. PMID- 16691104 TI - Changes in optical transmission caused by gamma ray induced coloring in photoluminescence dosimeter. AB - Transmission of visible light and ultraviolet radiation was examined for a phosphate-glass photoluminescence dosimeter irradiated with Co source gamma rays in the dose range of 1-60 Gy (H2O). The transmission for the wavelengths (lambda) less than 600 nm decreased with increasing irradiation dose beginning at 6 Gy. An approximate 20% reduction of transmission was observed for a 60 Gy exposure at the wavelength of ultraviolet radiation used for excitation (lambda = 337 nm). However, no change of transmission was seen in longer wavelength region (lambda > 600 nm), which includes the range of photoluminescence (lambda = 610-710 nm). Relative efficiencies of measured photoluminescence agreed well with estimations that were calculated from the transmission reduction of ultraviolet radiation. This fact indicates that reduction of photoluminescence efficiency induced by high-dose gamma rays is attributable mostly to attenuation of the ultraviolet radiation from an excitation source, rather than saturation of trapping or recombination centers. PMID- 16691105 TI - Childhood leukemia incidence and exposure to indoor radon, terrestrial and cosmic gamma radiation. AB - This study was undertaken to evaluate the ecological association between terrestrial and cosmic gamma radiation, indoor radon, and acute leukemia incidence among children under 15 y of age. From 1990 to 2001, 5,330 cases of acute leukemia were registered by the French National Registry of Childhood Leukemia and Lymphoma. Exposure to terrestrial gamma radiation was based on measurements, using thermoluminescent dosimeters, at about 1,000 sites covering all the "Departements." In addition, 8,737 indoor terrestrial gamma dose rate measurements covering 62% of the "Departements" and 13,240 indoor radon concentration measurements covering all the "Departements" were made during a national campaign. Cosmic ray doses were estimated in each of the 36,363 "Communes" of France. There was no evidence of an ecological association between terrestrial gamma dose (range: 0.22-0.90 mSv y) or total gamma dose (range: 0.49 1.28 mSv y) and childhood acute leukemia incidence, for acute myeloid leukemia (AML) or for acute lymphoblastic leukemia (ALL), in univariate or multivariate regression analyses including indoor radon. A significant positive association between indoor radon (range: 22-262 Bq m) and AML incidence among children was observed and remained significant in multivariate regression analyses including either terrestrial gamma dose [SIR per 100 Bq m = 1.29 (1.09-1.53)] or total gamma dose [SIR per 100 Bq m = 1.29 (1.09-1.53)]. The study showed no ecological association between terrestrial gamma radiation and childhood leukemia for the range of variation in gamma dose rates observed in France. The moderate ecological association between childhood AML incidence and indoor radon does not appear to be confounded by terrestrial gamma dose. PMID- 16691106 TI - Ancestry of beagles in lifespan studies of radionuclide toxicity at the University of Utah. AB - Analysis of the ancestry of the 1,262 lifespan beagles (LSB) entered into lifespan studies at the Radiobiology Division, University of Utah, beginning in 1952 and ending in 1980, indicated that about 97% of ancestor citations in the various pedigrees were of only 10 breeding animals (breeders) among breeders within the beagle colony. In turn, just 18 AKC-registered "champion" beagles from outside of this colony (founders) accounted for about 98% of all ancestor citations among founders for the LSB. We conclude from this study that the animals used in the lifespan radionuclide experiments can be considered to be somewhat genetically interrelated. PMID- 16691107 TI - Radiation doses to patients from extracorporeal shock wave lithotripsy. AB - The aim of this study was to determine the radiation doses to patients during extracorporeal shock wave lithotripsy (ESWL) and compare them with the available bibliographical data. In this method localization of the renal stones is attained by the use of fluoroscopy, and thus ESWL is included among those medical practices associated with patient radiation exposure. The entrance surface dose was measured using 3-4 thermoluminescent dosimeters positioned on the patients' backs at the entrance surfaces of the two x-ray beams for 50 ESWL procedures. Fluoroscopy time and number of spot films were also recorded. The average entrance surface dose at the patient's side with the renal stone was estimated to be 76.5 mGy for the oblique x-ray beam and 44.5 mGy for the PA x-ray beam. The mean fluoroscopy time was 204 s while 4 spot films were acquired on average. The mean effective dose (E) was estimated as 1.63 mSv per patient. The mean entrance surface dose values recorded in this study are comparable to but smaller than the values reported in the literature for ESWL, while the mean fluoroscopy time is within the range of values reported by other authors. On the other hand, the estimated E value is relatively higher compared to the corresponding values given in the literature based on patient measurements. However, it is comparable to recent published data acquired using extended measurements in an anthropomorphic phantom. PMID- 16691108 TI - Decommissioning procedures for an 11 MeV self-shielded medical cyclotron after 16 years of working time. AB - The present article describes the decommissioning of a compact, self-shielded, 11 MeV medical cyclotron. A Monte Carlo simulation of the possible nuclear reactions was performed in order to plan the decommissioning activities. In the course of the cyclotron dismantling, cyclotron components, shields, and floor concrete samples were measured. Residual activities were analyzed with a Ge(Li) detector and compared with simulation data. Doses to staff involved in the decommissioning procedure were monitored by individual TL dosimeters. The simulations identified five radioactive nuclides in shields and floor concrete: 55Fe and 45Ca (beta emitters, total specific activity: 2.29 x 10(4) Bq kg) and 152Eu, 154Eu, 60Co (gamma emitters, total specific activity: 1.62 x 10(3) Bq kg-1). Gamma-ray spectrometry confirmed the presence of gamma emitters, corresponding to a total specific activity of 3.40 x 10(2) Bq kg-1. The presence of the radioisotope 124Sb in the lead contained in the shield structure, corresponding to a simulated specific activity of 9.38 x 10(3) Bq kg-1, was experimentally confirmed. The measured dose from external exposure of the involved staff was <20 muSv, in accordance with the expected range of values between 10 and 20 muSv. The measured dose from intake was negligible. Finally, the decommissioning of the 11 MeV cyclotron does not represent a risk for the involved staff, but due to the presence of long-lived radioisotopes, the cyclotron components are to be treated as low level radioactive waste and stored in an authorized storage area. PMID- 16691112 TI - The pathogenesis of multiple sclerosis: relating human pathology to experimental studies. AB - Multiple sclerosis (MS) is a complex disease of unknown etiology. A careful study of the pathology of its component elements in relation to relevant experimental models has helped to understand some of the mechanisms that might be present in the disease. However, the autoimmune nature of the disease has recently come under question and there is a growing recognition of the importance of axonal, cortical, and white matter changes in the genesis and evolution of the lesions, their clinical diagnostic characteristics, and their response to treatment. This review highlights the emerging issues in MS from experimental, imaging and clinical perspectives. PMID- 16691113 TI - Atypical protein kinase C in neurodegenerative disease I: PKMzeta aggregates with limbic neurofibrillary tangles and AMPA receptors in Alzheimer disease. AB - Protein kinase Mzeta (PKMzeta), an atypical protein kinase C (PKC) isoform, plays a key role in the maintenance of long-term potentiation (LTP), a persistent enhancement of AMPA receptor-mediated synaptic transmission, as well as in the persistence of memory in Drosophila. Because memory impairment in Alzheimer disease (AD) has been attributed to disruption of synaptic plasticity, we investigated the expression and distribution of PKMzeta in this disorder. We found that PKMzeta accumulated in neurofibrillary tangles (NFTs), whereas conventional and novel PKC isoforms did not. Unlike tau, which is present in all NFTs regardless of location, PKMzeta was found in a subset of NFTs restricted to limbic or medial temporal lobe structures (i.e. hippocampal formation, entorhinal cortex, and amygdala), areas implicated in memory loss in AD. Interestingly, PKMzeta was not identified in any NFTs in control brains derived from 6 elderly individuals without known cognitive impairment. In medial temporal lobe structures in AD, PKMzeta also occurred within abnormal neurites expressing MAP2, GluR1 and GluR2 as well as in perisomatic granules expressing GluR1 and GluR2, suggesting that aggregation of PKMzeta disrupts glutamatergic synaptic transmission. Together, these findings suggest a link between PKMzeta-mediated synaptic plasticity and memory impairment in AD. PMID- 16691114 TI - Atypical protein kinase C in neurodegenerative disease II: PKCiota/lambda in tauopathies and alpha-synucleinopathies. AB - To study the role of atypical protein kinase C (aPKC) in neurodegenerative disease, we investigated the distribution of PKCiota/lambda, an aPKC isoform, in a variety of tauopathies and alpha-synucleinopathies. Immunohistochemical study revealed PKCiota/lambda within tau-positive neurofibrillary inclusions in Alzheimer disease (AD), progressive supranuclear palsy, corticobasal degeneration (CBD), and Pick disease (PiD), within alpha-synuclein-positive Lewy bodies in idiopathic Parkinson disease and dementia with Lewy bodies, as well as within glial inclusions in multisystem atrophy. We also observed PKCiota/lambda label of actin-rich Hirano bodies in AD, PiD, and elderly individuals. Double immunolabeling and fluorescence resonance energy transfer demonstrated close physical association between PKCiota/lambda and phospho-tau or alpha-synuclein in some neurofibrillary tangles and Lewy bodies. Furthermore, PKCiota/lambda colocalized with p62, a chaperone protein that binds to both aPKC and ubiquitin, in most of these inclusions. PKCiota/lambda also closely associated with the inactivated form of glycogen synthase kinase-3beta, GSK-3beta[ser9]. Together, these findings suggest that PKCiota/lambda may play a role in common mechanisms involving the pathogenesis of neurodegenerative disease. PMID- 16691115 TI - A possible cellular mechanism of neuronal loss in the dorsal root ganglia of Dystonia musculorum (dt) mice. AB - Dystonia musculorum (dt) is a mutant mouse with hereditary sensory neuropathy. A defective bullous pemphigoid antigen 1 (BPAG1) gene is responsible for this mutation. In the present study, we examined the distribution of neuronal intermediate filament proteins in the central and peripheral processes of the dorsal root ganglia (DRG) in adult dt mice using different approaches. We found that not only BPAG1, but also alpha-internexin was absent in the DRG neurons in adult dt mice. To study the relationship between the absence of alpha-internexin and the progressive neuronal loss in the DRG of dt mice, we further cultured DRG neurons from embryonic dt mutants. Immunocytochemical assay of cultured DRG neurons from dt embryos revealed that alpha-internexin was aggregated in the proximal region of axons and juxtanuclear region of the cytoplasma, yet the other intermediate filament proteins were widely distributed in all processes. The active caspase-3 activity was observed in the dt neuron with massive accumulation of alpha-internexin. From our observations, we suggest that the interaction between BPAG1 and alpha-internexin may be one of the key factors involved in neuronal degeneration, and abnormal accumulation of alpha-internexin may impair the axonal transport and subsequently turns on the cascade of neuronal apoptosis in dt mice. PMID- 16691116 TI - Endoplasmic reticulum stress features are prominent in Alzheimer disease but not in prion diseases in vivo. AB - Prion diseases and Alzheimer disease (AD) share a variety of clinical and neuropathologic features (e.g. progressive dementia, accumulation of abnormally folded proteins in diseased tissue, and pronounced neuronal loss) as well as pathogenic mechanisms like generation of oxidative stress molecules and complement activation. Recently, it was suggested that neuronal death in AD may have its origin in the endoplasmic reticulum (ER). Cellular stress conditions can interfere with protein folding and subsequently cause accumulation of unfolded or misfolded proteins in the ER lumen. The ER responds to this by the activation of adaptive pathways, which are termed unfolded protein response (UPR). The UPR transducer PERK, which launches the most immediate response to ER stress (i.e. the transient attenuation of mRNA translation), and the downstream effector of PERK, eIF2alpha, were shown to be activated in AD. We demonstrate that neither in sporadic nor in infectiously acquired or inherited human prion diseases can the activated forms of PERK and eIF2alpha be detected, except when concomitant neurofibrillary pathology is present; whereas the distribution of phosphorylated PERK correlates with abnormally phosphorylated tau in AD. In brains of scrapie affected mice and mice infected with sporadic or variant Creutzfeldt-Jakob disease, activated PERK is only very faintly expressed. The lack of prominent activation of the PERK-eIF2alpha pathway in prion diseases suggests that, in contrast to AD, ER stress does not play a crucial role in neuronal death in prion disorders. PMID- 16691117 TI - Transforming growth factor-beta1 is a negative modulator of adult neurogenesis. AB - Transforming growth factor (TGF)-beta1 has multiple functions in the adult central nervous system (CNS). It modulates inflammatory responses in the CNS and controls proliferation of microglia and astrocytes. In the diseased brain, TGF beta1 expression is upregulated and, depending on the cellular context, its activity can be beneficial or detrimental regarding regeneration. We focus on the role of TGF-beta1 in adult neural stem cell biology and neurogenesis. In adult neural stem and progenitor cell cultures and after intracerebroventricular infusion, TGF-beta1 induced a long-lasting inhibition of neural stem and progenitor cell proliferation and a reduction in neurogenesis. In vitro, although TGF-beta1 specifically arrested neural stem and progenitor cells in the G0/1 phase of the cell cycle, it did not affect the self-renewal capacity and the differentiation fate of these cells. Also, in vivo, TGF-beta1 did not influence the differentiation fate of newly generated cells as shown by bromo-deoxyuridine incorporation experiments. Based on these data, we suggest that TGF-beta1 is an important signaling molecule involved in the control of neural stem and progenitor cell proliferation in the CNS. This might have potential implications for neurogenesis in a variety of TGF-beta1-associated CNS diseases and pathologic conditions. PMID- 16691118 TI - Dystrophin expression in muscles of duchenne muscular dystrophy patients after high-density injections of normal myogenic cells. AB - A clinical trial was conducted to test a new protocol of normal muscle precursor cell (MPC) allotransplantation in skeletal muscles of patients with Duchenne muscular dystrophy (DMD). Cultured MPCs obtained from one of the patient's parents were implanted in 0.25 or 1 cm of a Tibialis anterior in 9 patients with DMD. MPC injections were placed 1 to 2 mm from each other, and a similar pattern of saline injections was done in the contralateral muscle. The patients were immunosuppressed with tacrolimus. Muscle biopsies were performed at the injected sites 4 weeks later. In the biopsies of the cell-grafted sites, there were myofibers expressing donor's dystrophin in 8 patients. The percentage of myofibers expressing donor's dystrophin varied from 3.5% to 26%. Evidence of small myofiber neoformation was observed in some patients. Donor-derived dystrophin transcripts were detected by reverse transcriptase-polymerase chain reaction in the cell-grafted sites in all patients. The protocol of immunosuppression was sufficient to obtain these results, although it is not certain whether acute rejection was efficiently controlled in all the cases. In conclusion, intramuscular allotransplantation of normal MPCs can induce the expression of donor-derived dystrophin in skeletal muscles of patients with DMD, although this expression is restricted to the sites of MPC injection. PMID- 16691119 TI - Lewy bodies in progressive supranuclear palsy represent an independent disease process. AB - Progressive supranuclear palsy (PSP) is a neurodegenerative tauopathy characterized by Parkinsonism, vertical gaze palsy, and early falls. Lewy bodies (LBs) are detected in approximately 10% of PSP cases, but there is little information on the relationship of LBs to tau pathology. We determined the frequency of LBs in a large series of autopsy-confirmed cases of PSP and studied the density and distribution of LBs, including Parkinson disease stage, in cases with LBs (PSP/LBD). PSP/LBD was compared with pure LB disease (LBD), including assessment of neuronal loss in key brainstem nuclei. Immunohistochemistry for alpha-synuclein revealed LBs in 31 of 290 PSP cases (11%). One case had multiple system atrophy in addition to PSP and was excluded from further study along with 2 PSP/LBD cases with concurrent Alzheimer disease. The 29 cases of PSP/LBD were compared with 30 cases of PSP and 24 cases of LBD. The age, sex, brain weight, Braak neurofibrillary tangle (NFT) stage, as well as counts of NFTs and senile plaques were not different among PSP, LBD, and PSP/LBD, but disease duration was longer in LBD. The Parkinson disease stage was similar, but the density of LBs in most subcortical nuclei tended to be greater in LBD than in PSP/LBD. In contrast, substantia nigra neuronal loss was greater in PSP/LBD than both PSP and LBD. Double immunostaining demonstrated alpha-synuclein and tau in different neurons with few exceptions. The findings suggest that LBs in PSP are similar in distribution to those in LBD and independent of tau pathology. The greater density of LBs in LBD compared with PSP/LBD may be the result of longer disease duration in LBD, whereas greater neuronal loss in the substantia nigra in PSP/LBD may be the result of vulnerability of this brain region to both disease processes. PMID- 16691120 TI - Atypical progressive supranuclear palsy with corticospinal tract degeneration. AB - Progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), sporadic multisystem tauopathy, and some forms of frontotemporal dementia with Parkinsonism linked to chromosome 17 are characterized by neuronal and glial lesions accumulating tau protein containing 4 conserved repeats in microtubule binding domain (4R tau). Corticospinal tract degeneration is not a common feature of 4R tauopathies. Our objective was to describe 12 cases with pathologic features similar to those of PSP but with prominent corticospinal tract degeneration. We reviewed the historical records and neuropathologic evaluation using standardized sampling, immunohistochemistry, semiquantitative analysis, image analysis, and electron microscopy. The mean age at onset and illness duration was 71 and 5.7 years, respectively. Eight cases were female. Eleven cases had clinical evidence of prominent upper motor neuron disease plus extrapyramidal features. There was focal parasagittal cortical atrophy involving motor cortex and degeneration of corticospinal tract with sparing of lower motor neurons like in primary lateral sclerosis. Prominent tau pathology was found in oligodendrocytes in motor cortex, subjacent white matter, and corticospinal tract characterized by globular cytoplasmic filamentous inclusions that were immunoreactive for 4R tau. The clinicopathologic features of these 12 cases expand the spectrum of 4R tauopathies. PMID- 16691121 TI - NG2 colocalizes with axons and is expressed by a mixed cell population in spinal cord lesions. AB - The NG2 proteoglycan is of general interest after spinal cord injury because it is expressed by oligodendrocyte progenitors (OPCs), which contribute to central nervous system remyelination; however, NG2 may inhibit axon regeneration. We and others have examined the spatiotemporal expression of NG2 after spinal cord injury (SCI). Here, we extend those observations and provide a comprehensive analysis of the distribution, phenotype, and colocalization of NG2 cells with axons in a clinically relevant model of spinal contusion. Because contusion models mimic the majority of human SCI, this information is important for understanding endogenous processes that promote and/or prevent repair. The data demonstrate that NG2 levels rise significantly between 3 and 7 days postinjury (dpi) and remain elevated chronically throughout the lesions. NG2 within the lesions could be derived from an array of infiltrating cells; thus, a panel of antibodies was used to investigate NG2 cell phenotypes. First, platelet-derived growth factor-alpha receptor (PDGFalphaR) colocalization was examined because OPCs normally express both markers. PDGFalphaR cells were present in lesions at all times examined. However, only 37% of NG2 cells coexpressed PDGFalphaR at 14 dpi, which dropped to <1% by 70 dpi. This contrasts with the nearly complete overlap in spared tissue surrounding the lesion. In contrast, 40% to 60% of NG2 cells expressed p75 and approximately 84% expressed Sox10, suggesting that many NG2 cells were nonmyelinating Schwann cells. Despite rising levels of NG2, we noted robust and sustained axon growth into the lesions, many of which were located along NG2 profiles. Thus, spinal contusion produces an NG2-rich environment into which axons grow and in which the source of NG2 appears considerably different from that in surrounding spared tissue. PMID- 16691124 TI - Meeting report for Molecular Psychiatry, 2005. AB - The 2005 Molecular Psychiatry Meeting was held from 6 to 8 February 2005 at the Treasure Mountain Inn in Park City, Utah. PMID- 16691125 TI - No association between genetic variants at the GLYT2 gene and bipolar affective disorder and schizophrenia. PMID- 16691126 TI - Lack of association between the corticotrophin-releasing hormone receptor 2 gene and panic disorder. AB - Panic disorder is classified as an anxiety disorder and affects 1-3% of the population. An individual suffering from such a disorder may experience unexpected recurrent panic attacks and fear of future attacks. Twin and family studies have pointed towards a strong heritability of the disorder. Stress response and anxiety are thought to be mediated, at least in part, by the corticotrophin-releasing hormone (CRH), which is known to be a regulator of the hypothalamic-pituitary-adrenal pathway. To search for markers conferring genetic susceptibility to panic disorder, we typed three polymorphisms of the CRHR2 gene CRHR2(CA), CRHR2(GT), and CRHR2(GAT) - in 466 individuals, 183 of whom had DSM IV panic disorder. Seventy-five case-controls and 101 triad families plus 13 siblings were examined. Case-control association analyses using chi(2) tests yielded no difference in the distribution of the alleles. Linkage analysis using the Transmission Disequilibrium Test showed no preferential transmission of alleles for any of the three markers. Haplotype analysis indicated that allele 7 of CRHR2 (GAT) and 8 of CRHR2 (GT) are in almost complete linkage disequilibrium (P = 0.000 000 1). Although both neurobiology and chromosomal location point to the CHRH2 receptor gene as a candidate for panic disorder, our study indicates that the CRHR2 polymorphisms examined do not confer susceptibility to panic disorder. Further studies investigating additional polymorphisms in this gene and other components of the CRH signalling system may prove useful. PMID- 16691127 TI - Genetics of panic disorder on the Faroe Islands: a replication study of chromosome 9 and panic disorder. AB - OBJECTIVE: The population of the Faroe Islands in the North Atlantic Ocean is likely to have the same ancestry as the Icelandic population. An Icelandic study on Panic Disorder has found some evidence for a loci on chromosome 9. METHODS: On the Faroe Islands we have an ongoing genetic project concerning panic disorder among other psychiatric disorders. We searched for shared alleles and haplotypes in distantly related cases from the isolated and recently found population of the Faroe Islands, using 26 more or less evenly distributed microsatellite markers on chromosome 9, with emphasis on the candidate region identified in the Icelandic study. RESULTS: We have not been able to replicate the Icelandic results. Owing to the study design and sample size, we would not be able to detect areas with small impact. PMID- 16691128 TI - C677T and A1298C methylenetetrahydrofolate reductase gene polymorphisms in schizophrenia, bipolar disorder and depression: a meta-analysis of genetic association studies. AB - A meta-analysis of the previous studies of allelic association between schizophrenia, bipolar disorder and depression with the C677T and A1298C polymorphisms of the methylenetetrahydrofolate reductase (MTHFR) gene was carried out. Attention was paid to genetic differences in ancestral groups. Ten studies of the C677T MTHFR polymorphism and schizophrenia were included, along with four studies with the A1298C MTHFR polymorphism and schizophrenia. Four association studies of the C677T MTHFR polymorphism with bipolar disorder, and five studies of the C677T base pair change with depression were included. The studies contained Caucasian and east Asian samples. For schizophrenia, the main analysis for investigating the association of the C677T allele T and the risk of developing schizophrenia relative to the allele C showed lack of heterogeneity (P = 0.22, I(2) = 24%) between samples. In these samples, the fixed effects odds ratio was marginally significant [odds ratio 1.13 (95% confidence interval, 1.04 1.23)]. In an analysis of subgroups, the odds ratio were not significant for the Caucasian sample [random effects odds ratio 1.14 (0.95-1.36) and fixed effects odds ratio 1.13 (1.00-1.27)](P = 0.14, I(2) = 43%) and in Asians [random effects odds ratio 1.15 (0.98-1.35) and fixed effects odds ratio 1.15 (1.01-1.31)] (P = 0.19, I(2) = 37). The recessive model for allele T produced significant results in the main analysis [fixed effects odds ratio 1.32 (1.12-1.56)] in east Asians [fixed effects odds ratio 1.45 (1.13-1.85)] and female participants [fixed effects odds ratio 1.70 (1.07-2.71)], whereas for Caucasians the results were nonsignificant. For the A1298C polymorphism, in the main analysis the allele frequency difference between C and A, and the dominant model for allele C produced marginally significant associations [fixed effects odds ratio 1.16 (1.03 1.31) and fixed effects odds ratio 1.19 (1.02-1.40), respectively]. In bipolar disorder, overall, there was no significant association between either allele of the C677T polymorphism and the risk of developing bipolar disorder. Only in the east Asian population was the C677T association of marginal significance, with fixed effects odds ratio 1.23 (1.00-1.52). No sources of potential bias were found in the selected studies. The meta-analysis results suggested that east Asians have a greater genetic risk from the MTHFR gene in developing schizophrenia and depression, and that the genetic effects in bipolar disorder and depression are different. A further exploration of the involvement of the MTHFR gene in the susceptibility to schizophrenia and affective disorders, with a greater number of studies with larger sample sizes, however, are needed to fully establish the role of the MTHFR gene. PMID- 16691129 TI - Catechol-O-methyltransferase Val158Met genotype variation is associated with prefrontal-dependent task performance in schizotypal personality disorder patients and comparison groups. AB - OBJECTIVE: A single-nucleotide polymorphism of the gene coding for catechol-O methyltransferase (COMT Val(158)Met) is associated with prefrontal-dependent task performance in schizophrenia. We evaluated the relationship of the COMT genotype with diagnostic status and cognitive performance in schizotypal personality disorder. METHODS: Unmedicated outpatients with schizotypal personality disorder (SPD; n = 67) and non-schizotypal personality disorder (NSPD; n = 154) by DSM-III R, and normal control (NC; n = 60) participants were genotyped at the COMT Val(158)Met locus. Of these, 98 Caucasians (23 SPD, 52 NSPD and 23 NC) performed a brief neurocognitive battery: Wisconsin Card Sorting Test (WCST), Paced Auditory Serial Addition Test (PASAT), California Verbal Learning Test (CVLT), Visuospatial Working Memory (DOT) and Visual Delayed Recall (Wechsler Memory Scale Visual Reproduction, WMS-VR). RESULTS: Allele distribution was not significantly different in the full sample (by chi(2)) for the SPD group compared with either the NC or combined NC/NSPD groups. In analyses of variance of Caucasian individuals, the SPD group performance met or approached significantly worse performance than NC, NSPD or both groups, on the PASAT, CVLT and WMS-VR. In regression analyses of cognitive performance, the COMT genotype was significantly associated with performance on WCST and PASAT, independent of diagnosis, with the Val/Val genotype associated with the worse performance. CONCLUSIONS: (1) Allelic variation in COMT activity is unrelated to the diagnosis of SPD in this sample. (2) Individuals with SPD exhibit multiple deficits in prefrontal and temporal lobe-dependent tasks. (3) The COMT genotype is related to performance on prefrontal cortex-dependent tasks and may contribute to the deficit in prefrontal dependent memory processes in SPD as it does in schizophrenia. PMID- 16691130 TI - Influence of the serotonin transporter gene on comorbid disorders among alcohol dependent individuals. AB - OBJECTIVE: The role of the human serotonin transporter protein (5-HTT) gene in psychiatric disorders suggests that its variation may influence the comorbidity pattern and the heterogeneity of alcoholism. The aim of the present study is to verify possible associations between the 5-HTTLPR control region polymorphism with alcoholism and comorbid disorders. METHODS: The polymorphic site was genotyped in 114 patients with alcohol dependence and 218 controls, all of them Brazilians of European descent. A comprehensive diagnostic interview identified the comorbid disorders. RESULTS: Study participants with alcohol dependence and controls did not differ in the genotype and allele frequencies (genotypes: chi(2) = 2.52, P = 0.28; alleles: chi(2) = 0.37; P = 0.54). Patients with comorbid major depressive disorder (chi(2) = 6.14, P = 0.01), drug abuse (chi(2) = 6.82, P = 0.01) and nicotine dependence (chi(2) = 4.10, P = 0.04), however, presented a higher frequency of the S allele than patients without these comorbidities. Patients with comorbid depression and drug abuse also presented a higher frequency of the S allele than controls. CONCLUSIONS: The present results are consistent with the importance of the 5-HTT gene in psychiatry. They suggest a role of the 5-HTTLPR polymorphism in a group of comorbid disorders among alcohol dependent individuals, supporting a genetic influence in alcoholism heterogeneity. PMID- 16691131 TI - Reliability and validity of the Pediatric Intensity Level of Therapy (PILOT) scale: a measure of the use of intracranial pressure-directed therapies. AB - OBJECTIVE: To test the reliability and validity of the Pediatric Intensity Level of Therapy (PILOT) scale, a novel measure of overall therapeutic effort directed at controlling intracranial pressure (ICP) in the setting of severe (Glasgow Coma Scale of A genotype to insulin resistance. AB - Type II diabetes is caused by a failure of the pancreatic beta-cells to compensate for insulin resistance leading to hyperglycaemia. There is evidence for an essential role of an increased beta-cell apoptosis in type II diabetes. High glucose concentrations induce IL-1beta production in human beta-cells, Fas expression and concomitant apoptosis owing to a constitutive expression of FasL. FASL and FAS map to loci linked to type II diabetes and estimates of insulin resistance, respectively. We have tested two functional promoter polymorphisms, FAS-670 G>A and FASL-844C>T as well as a microsatellite in the 3' UTR of FASL for association to type II diabetes in 549 type II diabetic patients and 525 normal glucose-tolerant (NGT) control subjects. Furthermore, we have tested these polymorphisms for association to estimates of beta-cell function and insulin resistance in NGT subjects. We found significant association to type II diabetes for the allele distribution of the FASL microsatellite (P-value 0.02, Bonferroni corrected). The FAS-670G>A was associated with homeostasis model assessment insulin resistance index and body mass index (P-values 0.02 and 0.02). We conclude that polymorphisms of FASL and FAS associate with type II diabetes and estimates of insulin resistance in Danish white subjects. PMID- 16691187 TI - Gene-expression profiling of HIV-1 infection and perinatal transmission in Botswana. AB - Perinatal transmission of human immunodeficiency virus (HIV)-1 represents a major problem in many regions of the world, especially Southern Africa. With the exception of viral and proviral load, the role for maternal cofactors in perinatal transmission outcome is largely unknown. In this study, an assessment was made of peripheral blood mononuclear cells (PBMC) gene-expression profiles to better understand transcriptional changes associated with HIV-1 infection and perinatal transmission among young adult mothers with infants in Botswana. Peripheral blood mononuclear cells specimens were used from 25 HIV+ drug naive and 20 HIV- healthy mothers, similar in age and location, collected in 1999-2000 and 2003, and processed with the exact same methods, as previously described. Expression profiling of 22 277 microarray gene probes implicated a broad initiation of innate response gene-sets, including toll-like receptor, interferon stimulated and antiviral RNA response pathways in association with maternal HIV-1 infection. Maternal transmission status was further associated with host genes that influence RNA processing and splicing patterns. In addition to real-time polymerase chain reaction validation of specific genes, enriched category validation of PBMC profiles was conducted using two independent data sets for either HIV-1 infection or an unrelated RNA virus, severe acute respiratory virus infection. HIV-1 pathogen-specific host profiles should prove a useful tool in infection and transmission intervention efforts worldwide. PMID- 16691188 TI - High-density SNP analysis of 642 Caucasian families with rheumatoid arthritis identifies two new linkage regions on 11p12 and 2q33. AB - We have completed a genome wide linkage scan using >5700 informative single nucleotide polymorphism (SNP) markers (Illumina IV SNP linkage panel) in 642 Caucasian families containing affected sibling pairs with rheumatoid arthritis (RA), ascertained by the North American Rheumatoid Arthritis Consortium. The results show striking new evidence of linkage at chromosomes 2q33 and 11p12 with logarithm of odds (LOD) scores of 3.52 and 3.09, respectively. In addition to a strong and broad linkage interval surrounding the major histocompatibility complex (LOD>16), regions with LOD>2.5 were observed on chromosomes 5 and 10. Additional linkage evidence (LOD scores between 1.46 and 2.35) was also observed on chromosomes 4, 7, 12, 16 and 18. This new evidence for multiple regions of genetic linkage is partly explained by the significantly increased information content of the Illumina IV SNP linkage panel (75.6%) compared with a standard microsatellite linkage panel utilized previously (mean 52.6%). Stratified analyses according to whether or not the sibling pair members showed elevated anticyclic citrullinated peptide titers indicates significant variation in evidence for linkage among strata on chromosomes 4, 5, 6 and 7. Overall, these new linkage data should reinvigorate efforts to utilize positional information to identify susceptibility genes for RA. PMID- 16691189 TI - Antiproliferative effect of semaphorin 3F on human melanoma cell lines. PMID- 16691190 TI - The EGF A61G polymorphism is associated with disease-free period and survival in malignant melanoma. AB - An earlier study reported that a common polymorphism in the 5' untranslated region of the epidermal growth factor (EGF) gene is associated with increased risk for cutaneous malignant melanoma (MM) and Breslow thickness. Since then, several independent studies have reported conflicting results that have challenged this hypothesis. However, none of the previous studies examined survival as the primary outcome. We therefore sought to study the association between this polymorphism and survival. One hundred and thirty patients diagnosed with MM with a Breslow thickness of >1.5 mm were included in this study. In our collective, the G/G genotype represented a significant risk factor for both shorter disease-free period (hazard ratio of 2.246, 95% CI: 1.06-4.78, P=0.036) and overall MM-specific survival (hazard ratio of 3.8, 95% CI: 1.5-9.5, P=0.004) compared with the A/A genotype, while the heterozygous A/G genotype showed an intermediate risk. In the present study, we demonstrate for the first time that the EGF A61G polymorphism is associated with survival. Our data suggest that this polymorphism is a potential marker for disease severity that predicts earlier progression of MM. PMID- 16691191 TI - Low-energy helium-neon laser induces locomotion of the immature melanoblasts and promotes melanogenesis of the more differentiated melanoblasts: recapitulation of vitiligo repigmentation in vitro. AB - Helium-neon laser (He-Ne Laser, 632.8 nm) is a low-energy laser that has therapeutic efficacy on various clinical conditions. Our previous study has demonstrated efficacy of He-Ne laser on vitiligo, a disease characterized by skin depigmentation. To regain skin tone on vitiligo lesions, the process began by the migration of the immature melanoblasts (MBs) to the epidermis, which was followed by their functional development to produce melanin. In this study, we investigated the physiologic effects of He-Ne laser irradiation on two MB cell lines: the immature NCCmelb4 and the more differentiated NCCmelan5. The intricate interactions between MBs with their innate extracelluar matrix, fibronectin, were also addressed. Our results showed that He-Ne laser irradiation enhanced NCCmelb4 mobility via enhanced phosphorylated focal adhesion kinase expression and promoted melanogenesis in NCCmelan5. In addition, He-Ne laser decreased the affinity between NCCmelb4 and fibronectin, whereas the attachment of NCCmelan5 to fibronectin increased. The alpha5beta1 integrin expression on NCCmelb4 cells was enhanced by He-Ne laser. In conclusion, we have demonstrated that He-Ne laser induced different physiologic changes on MBs at different maturation stages and recapitulated the early events during vitiligo repigmentation process brought upon by He-Ne laser in vitro. PMID- 16691192 TI - Both pimecrolimus and corticosteroids deplete plasmacytoid dendritic cells in patients with atopic dermatitis. PMID- 16691193 TI - High frequency of BRAFV600E mutation in acquired nevi and small congenital nevi, but low frequency of mutation in medium-sized congenital nevi. AB - To investigate whether the frequency of the BRAF(V600E) (V-raf murine sarcoma virus oncogene homolog B1) mutation in melanocytic nevi is associated with sun exposure patterns, we examined 120 acquired melanocytic nevi excised from various anatomic sites, including glabrous skin, as well as 62 congenital nevi. We used a new mutation detection system based on the shifted termination assay, called Mutector, which was able to detect only 5% of heterozygous mutant cells within the samples. We detected the mutation in 105/120 (87.5%) acquired nevi and 43/62 (69.4%) congenital nevi. Notably, we found the mutation in 35/43 (81.4%) acquired nevi excised from glabrous skin and genitalia. These results strongly suggest that UV light is not necessarily required for the acquisition of the BRAF(V600E) mutation, and suggest that non-mutagenic effects of UV light to melanocytes may be more important in the nevogenesis. Additionally, we showed heterogeneous distribution of BRAF-mutated cells within the lesions of small congenital nevi by a combination of laser microdissection and direct sequencing. Finally, we found low frequency of BRAF(V600E) mutation (6/20, 30.0%) in medium-sized congenital nevi. Most of these nevi with wild-type BRAF had neroblastoma ras viral oncogene homolog mutations (9/14, 64.3%), suggesting different pathogenesis of medium sized congenital nevi from acquired nevi and small congenital nevi. PMID- 16691194 TI - Ameliorating skin-homing receptors on malignant T cells with a fluorosugar analog of N-acetylglucosamine: P-selectin ligand is a more sensitive target than E selectin ligand. AB - Expression of E- and P-selectin ligands is required for T cell entry into skin. Sialyl Lewis X moieties are critical for ligand activity and are elevated on malignant skin-homing T cells. We hypothesize that these glycosylations are selectable targets for treating the dermal tropism associated with cutaneous lymphomas. In this study, we analyzed the efficacy of a novel 4-fluorinated analog of N-acetylglucosamine (GlcNAc) on E- and P-selectin ligands expressed by malignant skin-homing T cells. We also examined the specificity of 4-F-GlcNAc (2 acetamido-1,3,6-tri-O-acetyl-4-deoxy-4-fluoro-D-glucopyranose) action by contrasting the effects on sialyl Lewis X expression displayed by P-selectin glycoprotein ligand-1 (PSGL-1) with sialylated O-glycans expressed by CD43. Using parallel-plate flow analysis, we found that 4-F-GlcNAc elicited 5-fold more potent inhibition on P-selectin ligand activity than on E-selectin ligand activity. To determine whether glycosylations conferring E- and P-selectin ligand activities were inhibited, we analyzed the expression of sialyl Lewis X and sialyl-fucosylated core 2 O-glycan (CHO-131 antigen), respectively. We found that 4-F-GlcNAc treatment resulted in dose-dependent ablation of sialyl Lewis X and CHO-131 antigen expression on PSGL-1, whereas sialylated O-glycans on CD43 were minimally affected. These results indicate that 4-F-GlcNAc treatment can selectively downregulate the P-selectin ligand activity and potentially prevent dermal dissemination of cutaneous lymphomas. PMID- 16691195 TI - Identification of a novel locus for progressive symmetric erythrokeratodermia to a 19.02-cM interval at 21q11.2-21q21.2. PMID- 16691196 TI - Serine protease signaling of epidermal permeability barrier homeostasis. AB - Evidence is growing that protease-activated receptor-2 (PAR-2) plays a key role in epithelial inflammation. We hypothesized here that PAR-2 plays a central role in epidermal permeability barrier homeostasis by mediating signaling from serine proteases (SP) in the stratum corneum (SC). Since the SC contains tryptic- and chymotryptic-like activity, we assessed the influence of SP activation/inhibition on barrier function. Acute barrier disruption increases SP activity and blockade by topical SP inhibitors (SPI) accelerates barrier recovery after acute abrogation. This improvement in barrier function is due to accelerated lamellar body (LB) secretion. Since tryptic SP signal certain downstream responses through PAR-2, we assessed its potential role in mediating the negative effects of SP on permeability barrier. Firstly, PAR-2 is expressed in the outer nucleated layers of the epidermis and most specifically under basal condition to the lipid raft (LR) domains. Secondly, tape stripping-induced barrier abrogation provokes PAR-2 activation, as shown by receptor internalization (i.e. receptor movement from LR to cytolpasmic domains). Thirdly, topical applications of PAR-2 agonist peptide, SLIGRL, delay permeability barrier recovery and inhibit LB secretion, while, conversely, PAR-2 knockout mice display accelerated barrier recovery kinetics and enhanced LB secretion, paralleled by increased LR formation and caveolin-1 expression. These results demonstrate first, the importance of SP/SPI balance for normal permeability barrier homeostasis, and second, they identify PAR-2 as a novel signaling mechanism of permeability barrier, that is, of response linked to LB secretion. PMID- 16691197 TI - Transforming growth factor-alpha: a major human serum factor that promotes human keratinocyte migration. AB - In unwounded skin, human keratinocytes (HKs) are in contact with a plasma filtrate. In an acute wound, HKs come in contact with serum for the first time. Because human serum (HS), but not plasma, promotes HK migration, we speculated that a major HK pro-motility factor in vivo comes from serum. In this study, we compared all of the published growth factors (GFs), reported to promote HK migration, with HS. No single GF could duplicate the HK pro-motility activity in HS. Among these GFs, transforming growth factor-alpha [corrected] showed the highest HK pro-motility activity, reaching approximately 80% of the activity in HS. The order of potency was: TGFalpha > insulin > EGF > heparin binding (HB)-EGF > IGF-1 > basic fibroblast growth factor >IL-8 > HGF > IL-1 > KGF>TGFbeta. Interestingly, the combination of TGFalpha and insulin could duplicate the HK pro motility activity in HS, although only the TGFalpha, but not insulin, levels increase in serum over plasma. Addition of neutralizing antibodies against TGFalpha to serum or depletion of TGFalpha from serum by immunoprecipitation significantly abolished its HK pro-motility activity. Plasma with added TGFalpha stimulated HK migration that reached more than 80% of the serum stimulation. Since insulin levels are identical between plasma and serum, we propose that TGFalpha is the physiologic HK pro-motility factor in HS. PMID- 16691198 TI - A metabolic enzyme of the short-chain dehydrogenase/reductase superfamily may moonlight in the nucleus as a repressor of promoter activity. AB - Transcriptional repression often depends on the action of recruited co-repressor complexes with intrinsic enzymatic activities. The composition of these complexes depends on the nicotine amide dinucleotide co-factors and is thus directly reflective of the metabolic state of the cells. This study provides evidence that an enzyme, hRoDH-E2, with cytoplasmic phosphorylated and reduced forms of NAD dependent retinol dehydrogenase activity may function in the nucleus as a transcriptional repressor. By using the promoter of the epidermal late differentiation marker profilaggrin as a model, we show that both in vivo and in vitro the protein is recruited over the promoter. hRoDH-E2 represses profilaggrin promoter activity by altering the function of other activators, such as Sp1. The repressive function is associated with the ability of nuclear hRoDH-E2 to modulate the acetylation/deacetylation activity in the vicinity of transcription initiation site. These findings add hRoDH-E2 to the small group of metabolic enzymes, which, by being recruited over promoter regions, could directly link the cytoplasmic and nuclear functions within the cell. PMID- 16691199 TI - The modulation of aromatase and estrogen receptor alpha in cultured human dermal papilla cells by dexamethasone: a novel mechanism for selective action of estrogen via estrogen receptor beta? AB - Steroid hormones have important modulatory effects on the hair follicle, but the mechanisms by which they regulate human hair growth are still poorly understood. It is now clear that there are two distinct estrogen receptors (estrogen receptor alpha (ERalpha) and estrogen receptor beta (ERbeta)) that bind 17beta-estradiol. Since the follicular dermal papilla is known to control hair growth, and steroid hormones regulate receptor and aromatase expression in other tissues, we tested the hypothesis that steroid hormones would similarly modulate estrogen receptor and/or aromatase expression in cultured dermal papilla cells derived from human hair follicles. Primary cultures of non-balding occipital and frontal scalp and beard dermal papilla cells (n = 10) were established. Immunocytochemical studies showed the expression of ERalpha in both the cytoplasm and nucleus, whereas ERbeta was confined to the nuclei. The cells derived from occipital scalp were also incubated for 24 hours with 10 nM of either 17beta-estradiol, estrone, testosterone, 5alpha-dihydrotestosterone, 5alpha-androstane-3alpha, 17beta-diol, 5alpha-androstane-3beta, 17beta-diol, or 100 nM tamoxifen or dexamethasone in phenol red-free, serum-free medium to measure the steady-state levels of ERalpha, ERbeta, and aromatase mRNA by semiquantitative reverse transcriptase-PCR. Although androgens and estrogens did not alter ERalpha mRNA levels, treatment with dexamethasone significantly reduced ERalpha levels to 38% of the untreated control. By contrast, ERbeta mRNA levels were unaffected by any steroid treatment. Furthermore, dexamethasone significantly stimulated the expression of aromatase mRNA approximately 9-fold. Aromatase activity, assayed by the tritiated water method, was stimulated in both frontal scalp and beard dermal papilla cell cultures by dexamethasone. These observations provide evidence for a glucocorticoid-dependent mechanism whereby the selective action of estradiol via ERbeta may be promoted. Additionally, upregulation of aromatase combined with downregulation of ERalpha provides a basis for selective action of estradiol produced locally by autocrine or paracrine mechanisms. PMID- 16691200 TI - Production of low titers of anti-desmoglein 1 IgG autoantibodies in some patients with staphylococcal scalded skin syndrome. PMID- 16691201 TI - Diminished induction of skin fibrosis in mice with MCP-1 deficiency. AB - Scar and fibrosis are often the end result of mechanical injury and inflammatory diseases. One chemokine that is repeatedly linked to fibrotic responses is monocyte chemoattractant protein-1 (MCP-1). We utilized a murine fibrosis model that produces dermal lesions similar to scleroderma to evaluate collagen fibrillogenesis in the absence of MCP-1. Dermal fibrosis was induced by subcutaneous injection of bleomycin into the dorsal skin of MCP-1-/- and wild type C57BL/6 mice. After 4 weeks of daily injections, bleomycin treatment led to thickened collagen bundles with robust inflammation in the lesional dermis of wild-type mice. In contrast, the lesional skin of MCP-1-/- mice exhibited a dermal architecture similar to phosphate-buffered saline (PBS)-injected control and normal skin, with few inflammatory cells. Ultrastructural analysis of the lesional dermis from bleomycin-injected wild-type mice revealed markedly abnormal arrangement of collagen fibrils, with normal large diameter collagen fibrils replaced by small collagen fibrils of 41.5 nm. In comparison, the dermis of bleomycin-injected MCP-1-/- mice displayed a uniform pattern of fibril diameters that was similar to normal skin (average diameter 76.7 nm). The findings implicate MCP-1 as a key determinant in the development of skin fibrosis induced by bleomycin, and suggest that MCP-1 may influence collagen fiber formation in vivo. PMID- 16691203 TI - Sharing science. PMID- 16691202 TI - Homozygous missense mutation in fibulin-5 in an Iranian autosomal recessive cutis laxa pedigree and associated haplotype. AB - Cutis laxa is a rare group of inherited and acquired disorders characterized by loose and redundant skin with reduced elasticity. Mutations in the elastin coding gene have been shown to cause autosomal dominant cutis laxa in three families. A homozygous mutation in the fibulin-5 coding gene was discovered in a Turkish pedigree showing recessive inheritance, and a different mutation in this gene was found in the heterozygous state in a sporadic case of the disease. Here, we report the third case of a mutation in the fibulin-5 coding gene in a recessive Iranian cutis laxa pedigree. The mutation is the same as previously reported in the Turkish pedigree, further confirming that it is causative of disease. A haplotype consisting of seven intragenic sequence variations common to both pedigrees is described for the mutation-carrying fibulin-5 allele. PMID- 16691204 TI - The ABCs of centromeres. PMID- 16691205 TI - Protein expression: one by one. PMID- 16691206 TI - Pore-ing the right dose. PMID- 16691207 TI - Filling the GAP for dynamin. PMID- 16691208 TI - Separating sisters. PMID- 16691209 TI - The permeability transition and BCL-2 family proteins in apoptosis: co conspirators or independent agents? PMID- 16691210 TI - White adipocytes are less prone to apoptotic stimuli than brown adipocytes in rodent. PMID- 16691211 TI - The protease inhibitor Ucf-101 induces cellular responses independently of its known target, HtrA2/Omi. PMID- 16691212 TI - Identification of a 'genuine' mammalian homolog of nematodal CED-4: is the hunt over or do we need better guns? PMID- 16691213 TI - ATP-binding cassette transporter 1 and transglutaminase 2 act on the same genetic pathway in the apoptotic cell clearance. PMID- 16691214 TI - The role of outcomes research in improving patient care. PMID- 16691215 TI - Urethral bulking agents. PMID- 16691216 TI - Treatment of kidney stones: current lithotripsy devices are proving less effective in some cases. PMID- 16691232 TI - Does gonadotropin-releasing-hormone agonist therapy for prostate cancer increase the risk of fracture? PMID- 16691233 TI - A risk-adapted approach to the treatment of stage I seminoma. PMID- 16691234 TI - Retroperitoneal lymph-node dissection after chemotherapy for germ cell cancer in patients with elevated tumor markers. PMID- 16691235 TI - Can PSA nadir predict prognosis in men receiving hormonal therapy for prostate cancer after primary treatment failure? PMID- 16691236 TI - Transurethral resection of superficial bladder cancer using 5-aminolevulinic acid: are there any long-term benefits? PMID- 16691237 TI - Dose escalation with proton-beam boost for low-risk prostate cancer. PMID- 16691238 TI - Do prostate cancer patients at risk of pelvic lymph-node metastasis benefit from pelvic radiation? PMID- 16691239 TI - Testosterone replacement therapy for late-onset hypogonadism. AB - Cross-sectional and longitudinal studies have shown that testosterone levels fall with increasing age. Low testosterone levels are reported in 20% of men aged 60 69 years, and 30% of men in this age-group are thought to have low free testosterone levels. The prevalence of low testosterone levels increases in older men and, therefore, as the average age of men in developed countries is increasing, there will be even more hypogonadal men in the future. Many of the symptoms and signs associated with testosterone deficiency in younger men are present in aging men. Several small studies suggest that testosterone replacement therapy (TRT) can be beneficial for some aging men, but men in these studies have not been uniformly testosterone-deficient, and responses have been inconsistent. TRT might be more beneficial and safer in men aged below 65 years than in men aged 65 years or older. There is a need for large clinical trials in both age groups to provide more definitive information for patients and clinicians as to the safety and efficacy of TRT. Before such data are available, clinicians must be cautious in offering these treatments to aging men. Close monitoring of patients undergoing testosterone supplementation is essential. PMID- 16691240 TI - Primer: tissue fixation and preservation for optimal molecular analysis of urologic tissues. AB - Appreciation of the different methods of tissue handling is a prerequisite to obtaining accurate and biologically relevant tissue-based information. When a tissue sample is removed from its environment, biological changes are induced within its constituent cell population. It is inevitable that artefacts will be induced through obtaining and processing tissues, irrespective of whether the samples comprise a few cells derived by fine-needle aspiration or larger specimens obtained surgically. Depending upon the level of sophistication of the analytical methods subsequently employed, such changes might be irrelevant, or might result in acquisition of spurious data. While even brief ischemia alters expression of some genes, detectable by appropriate molecular techniques, the same changes might make no appreciable difference to tissue histomorphology. Furthermore, the phenotype of viable cells is known to change during tissue collection and handling. For example, transitional epithelial cells voided in urine are not phenotypically identical to those retained within the urothelium. Such phenotypic changes are temporary and might be of little consequence to subsequent analyses. Surprisingly, many cells in tissues preserved in an ischemic state can remain viable for several hours, and are believed to remain genotypically stable in the short term. PMID- 16691241 TI - Renal medullary carcinoma in a patient with sickle-cell disease. AB - BACKGROUND: A 36-year-old black male with sickle-cell disease, asthma, and dyslipidemia presented with shortness of breath, chest pain, lethargy, and recent fever. Physical examination revealed mild respiratory distress. INVESTIGATIONS: Chest X-ray, CT, abdominal ultrasound, and MRI. DIAGNOSIS: Renal medullary carcinoma. PMID- 16691242 TI - Management of panurothelial disease in superficial bladder cancer. AB - BACKGROUND: A 59-year-old female presented with a history of recurrent superficial bladder cancer. Since the initial diagnosis 12 years earlier, she had had 12 recurrences, all treated with transurethral resection with adjuvant bacillus Calmette-Guerin and mitomycin C. Pathology samples had always shown Ta grade 1 transitional-cell carcinoma. At the last recurrence, she presented with several papillary lesions throughout the bladder, discovered on control cystoscopy. Transurethral resection revealed a papillary grade 3 transitional cell carcinoma of the bladder. INVESTIGATIONS: Intravenous urography and ureteroscopy. DIAGNOSIS: Panurothelial superficial transitional-cell carcinoma. MANAGEMENT: Radical cystectomy with total right ureterectomy and distal left ureterectomy. Reconstruction of the urinary tract was performed with the distal ileum detubulized and rearranged in a U shape. A pyeloileal anastomosis and a proximal ureteroileal anastomosis were performed at the right and left sides respectively. PMID- 16691243 TI - The SIRP family of receptors and immune regulation. AB - The immune system must be highly regulated to obtain optimal immune responses for the elimination of pathogens without causing undue side effects. This tight regulation involves complex interactions between membrane proteins on leukocytes. Members of the signal-regulatory protein (SIRP) family, which are expressed mainly by myeloid cells, provide one example of these regulatory membrane proteins. There are three SIRP-family genes that encode proteins that have similar extracellular regions but different signalling potentials, and are therefore known as 'paired receptors'. In this Review, we describe recent studies defining the ligands of the SIRP-family members, with particular emphasis on relating the molecular interactions of these proteins to their role in immune cell regulation. PMID- 16691244 TI - Dendritic cells in a mature age. AB - A common view supposes that dendritic cells (DCs) exist in two basic functional states: immature DCs induce tolerance to self, whereas mature DCs induce immunity to foreign antigens. However, the term 'mature' is often used not only functionally to designate immunogenic DCs but also as a phenotypic description of DCs expressing high levels of MHC, adhesion and co-stimulatory molecules. The recent realization that DCs can express such markers under non-immunogenic conditions raises the question of whether the two connotations of the term 'mature' should continue to be used interchangeably. Here, I discuss the origins of the maturation model and how terminology is evolving to better accommodate our current understanding of the function of DCs. PMID- 16691245 TI - Therapeutic deep lamellar keratoplasty for corneal perforations. AB - OBJECTIVES/AIMS: Corneal perforation can be potentially blinding unless the integrity of the globe is restored quickly. Although penetrating keratoplasty (PK) may achieve this, it carries a high risk of endothelial rejection in inflamed eyes. Deep lamellar keratoplasty (DLK) may be an alternative option to PK in such eyes owing to its potential for a lower incidence of rejection. We report the efficacy of DLK in patients with corneal perforations. PATIENTS AND METHODS: Four patients underwent layer-by-layer DLK for noninfective corneal perforation, after measures such as the use of a bandage contact lens, tissue adhesive, and conjunctival pedicle flap had failed. The preoperative visual acuity was hand movements in one patient, 1/60 in two, and 6/60 in one. All four had iris incarcerated within the corneal perforations. SF6 gas (three patients) and air (one patient) were injected into the anterior chamber at the end of surgery. RESULTS: The integrity of the globe was restored in all four patients with an improvement in visual acuity (6/60 in one and 6/36 or better in three). The mean follow-up time was 7 months. All four patients had clear corneas 3 months postoperatively, apart from the area of the original perforation. There was no recurrence of ulceration or perforation. CONCLUSION: DLK is a safe and effective therapeutic measure in the management of patients with corneal perforations acting to preserve the integrity of the globe and restore vision. PMID- 16691246 TI - Ocular gunpowder injury. PMID- 16691247 TI - Benign episodic unilateral mydriasis. PMID- 16691248 TI - Late-onset capsular block syndrome without lens displacement. PMID- 16691249 TI - A case of endogenous Candida albicans endophthalmitis resolving with itraconazole treatment without recourse to vitrectomy. PMID- 16691250 TI - Transient formed visual hallucinations following macular hole surgery. PMID- 16691251 TI - Prognostic factors for visual acuity improvement after intravitreal triamcinolone injection. AB - PURPOSE: In some patients with macular oedema, intravitreal triamcinolone acetonide injection (IVTA) fails to improve visual acuity, although oedema shows clinical and angiographic improvement. Side effects can include increased intraocular pressure, cataract development, and (rarely) endophthalmitis. Our purpose was to identify prognostic factors for visual acuity improvement after IVTA. METHODS: Data on patients treated by IVTA for macular oedema were retrospectively reviewed. Three months postinjection, visual acuity was rated as 'improved' (two or more Snellen lines gained) or 'nonimproved' (unchanged or worsened). Comparative demographic data and pre- and post-IVTA clinical and fluorescein angiographic findings were analysed with SPSS software. RESULTS: Of 57 eyes (57 patients), 27 (47%) improved after IVTA. Initial visual acuity ('good', 'moderate', or 'poor') and aetiology of macular oedema (diabetic, venous occlusion, or pseudophakic) did not differ between the two groups. Improvement occurred in significantly more eyes with clinical or angiographic evidence of cystoid macular oedema (CME) than in those with diffuse retinal thickening (P=0.04) or diffuse leakage on fluorescein angiography (P=0.02), respectively, and in significantly more pseudophakic than phakic eyes (P=0.046). CONCLUSIONS: Pseudophakia and clinical or angiographic CME, but not aetiology or initial visual acuity, were prognostic of visual acuity improvement after IVTA for macular oedema. PMID- 16691252 TI - Traumatic wound dehiscence after penetrating keratoplasty-a cause for concern. AB - AIM: We report the incidence, causes, characteristics, and the outcome of traumatic corneal graft ruptures in a tertiary referral centre in the UK. METHOD: A retrospective analysis of all graft ruptures secondary to trauma that were treated at our centre between 1999 and 2005 was undertaken. Statistical analysis of possible prognostic factors was undertaken using the Fisher's test. RESULTS: Nineteen eyes in 18 patients sustained eye trauma resulting in graft rupture (median age of 53 years; range 27-82; 15 men and four women). Seventeen cases were accidental and two were from violence. The median time interval between grafting and rupture was 8.3 months (range 3 days to 15 years). The 6-year incidence was 3.8%. All graft ruptures occurred at the host-graft junction and ranged from 45 to 270 degrees . Iris prolapse/loss was noticed in 89% and lens loss in 53%. The most common posterior segment complication was vitreous loss (74%), followed by vitreous haemorrhage (32%) and retinal detachment (21%). Grafts with 180 degrees or more of dehiscence were more likely to fail (P<0.001), had more extensive posterior segment damage, and a poorer visual outcome. Grafts without sutures had a more extensive dehiscence (P<0.01). Final visual acuity was worse than 6/60 in 58%. CONCLUSIONS: The risk of traumatic corneal graft rupture is significant and is associated with a poor visual outcome. This fact needs to be clearly emphasised during preoperative counselling and protective measures encouraged. PMID- 16691253 TI - Use of the operating microscope for scleral buckling. PMID- 16691254 TI - Advantages of modular phacoemulsification training. PMID- 16691255 TI - Characteristics of primary rhegmatogenous retinal detachment in Taiwan. AB - PURPOSE: To investigate the epidemiological characteristics and related risk factors for primary rhegmatogenous retinal detachment (RRD) in Taiwan. METHODS: The case-control study was based on retrospective chart review of hospital patients treated for primary RRD from 1995 to 2001, inclusively. The preoperative fundus findings and refractive status were collected for each patient. Controls were selected from a nationwide survey of visual impairment in the adult population during the same period. Risk factors for RRD were analysed by logistic regression. A total of 1032 RRD cases and 3537 controls were enrolled for the study. RESULTS: A pronounced bipeak pattern was evident in the age distribution for primary RRD in the third and sixth decades of life. Atrophic hole with lattice degeneration was preferential to younger (20-30 years) and highly myopic individuals (-7.4+/-5 D), whereas the flap tear tended to occur in middle-aged individuals (50-60 years) and those with moderate myopia (-4.1+/-5 D). The odds ratio for primary RRD with myopia, male gender, and older age (>40 years) were 1.33/D, 2.15, and 1.69, respectively. CONCLUSIONS: Myopia is an important RRD risk factor for young Taiwanese. The increasing prevalence of myopia has predisposed the young population to RRD. PMID- 16691256 TI - A comparison of HRT II and GDx imaging for glaucoma detection in a primary care eye clinic setting. AB - PURPOSE: To evaluate the performance of the HRT II (Heidelberg retinal tomograph) and GDx (glaucoma detection) retinal nerve fibre analyzer in GDx when used in the primary care eye clinic setting for glaucoma screening. PATIENTS AND METHODS: The study was prospective, cross-sectional, and hospital-based. One-hundred and twelve patients, 59 women and 53 men with a mean age of 57.8 years (range 18-85 years), had consecutive HRT II disc imaging and GDx retinal nerve fiber layer analysis. The Moorfield's regression classification and the 'GDx number' were used to predict the likelihood of glaucoma. A separate clinician, masked to the instrument results determined a definitive diagnosis, based on clinical examination. The extent of agreement between instrument prediction and the clinician diagnosis of glaucoma was examined by generating sensitivity and specificity tables. RESULTS: The HRT II had a sensitivity of 0.79 (95% CI: 0.60 0.92) and a specificity of 0.70 (95% CI: 0.60-0.78). The positive predictive value of the HRT II was 0.43 (95% CI: 0.29-0.57). Using a GDx number of 50 as 'cutoff' for glaucoma detection, the GDx had a sensitivity of 0.80 (95% CI: 0.59 0.93) and a specificity of 0.72 (95% CI: 0.61-0.80), with a positive predictive value of 0.43 (95% CI: 0.28-0.59). CONCLUSIONS: For glaucoma detection, neither the HRT II nor the GDx are effective as stand-alone screening devices in the primary care setting. PMID- 16691257 TI - Phacoemulsification with nondominant hand. AB - PURPOSE: Evaluation of on axis phacoemulsification surgery through temporal incision using nondominant hand with surgeon sitting at the head end, inpatients with against-the-rule astigmatism. METHODS: Eighty eyes of 80 patients who underwent phacoemulsification through a temporal clear corneal tunnel for age related cataract and against-the-rule astigmatism were enrolled and divided into four equal groups. In Group 1A, the surgeon was sitting at the head end for the left eye performing surgery with the left hand (nondominant hand). In group 1B, the surgeon was seated at the temporal side and surgery was performed in the left eye with dominant right hand. In group 2A, the surgeon was sitting at the head end for the right eye and performed surgery holding the phacoemulsification hand piece in his right hand. In group 2B, the surgeon sat on the temporal side of the right eye and performed phacoemulsification with his right hand. The patients were followed up on day 7, 1 month, and 3 months. Parameters evaluated included average phaco power, effective phaco time, uncorrected and best-corrected visual acuity, keratometry, intraocular pressure, surgically induced astigmatism, pachymetry, and endothelial cell counts. RESULTS: The phaco time and phaco power among the four groups were comparable (phaco time: P=0.368; phaco power: P=0.294). The four groups were also comparable on parameters like surgically induced astigmatism (P=0.674), change in postoperative keratometric astigmatism (P=0.584), endothelial cell loss (0.921), change in ultrasonic pachymetry (P=0.476), and intraocular pressure (P=0.942). No intraoperative or postoperative complications were observed in any of the groups. The mean uncorrected visual acuity at 3 months in group 1 was 0.723+/-0.21; in group 2 it was 0.756+/-0.21; in group 3 it was 0.748+/-0.22, and in group 4 it was 0.732+/-0.23. The best corrected visual acuity was 0.96+/-0.10, 0.97+/-0.11, 0.95+/-0.13, and 0.96+/ 0.10 in the four groups at 3 months. CONCLUSION: Phacoemulsification surgery can be successfully performed with nondominant hand with a good surgical outcome. The technique gives an alternative approach where surgeon does not have to shift the position to perform on-axis phacoemulsification. PMID- 16691258 TI - Intrableb triamcinolone acetonide injection after bleb-forming filtration surgery (trabeculectomy, phacotrabeculectomy, and trabeculectomy revision by needling): a pilot study. PMID- 16691259 TI - Argon laser photocoagulation for diabetic macular oedema. PMID- 16691260 TI - Panophthalmitis secondary to infection with Citrobacter koseri. PMID- 16691261 TI - Sympathetic ophthalmia following severe fungal keratitis. PMID- 16691262 TI - Physiological changes during hemodialysis in patients with intradialysis hypertension. AB - Intradialysis hypertension is a frustrating complication among hemodialysis (HD) patients. This study was conducted to investigate the physiological changes during intradialytic hypertension. The beat-to-beat continuous heart rate, hematocrit (Hct) changes during HD, serum levels of nitric oxide, plasma levels of catecholamine, renin, endothelin (ET-1), cardiac output (CO), and peripheral vascular resistance (PVR) were measured before and after HD in patients prone to develop intradialysis hypertension (n = 30) and from age, sex-matched control HD subjects (n = 30). It was found that the baseline values of Hct, serum levels of nitric oxide, plasma levels of catecholamine, renin, and ET-1, CO, PVR, and power index (low frequency/high frequency ratios) of heart rate variability were not significantly different between the patients and control subjects. In the hypertension-prone group, the plasma levels of catecholamine, renin, and the serial measurements of power index, did not show significant changes. However, the patients showed a significant elevation of systemic vascular resistance (56.8 +/- 9.2% vs 17.7 +/- 9.5; P < 0.05), ET-1 (510.9 +/- 43.3 vs 276.7 +/- 30.1 pg/ml; P < 0.05) and a significant decrease of nitric oxide (NO)/ET-1 balance (0.018 +/- 0.003 vs 0.034 +/- 0.005; P < 0.05) at the end of HD compared with the control patients. It was found that the physiological changes in intradialysis hypertension patients were characterized by inappropriately increased PVR through mechanisms that did not involve sympathetic stimulation or renin activation but might be related with altered NO/ET-1 balance. PMID- 16691264 TI - [Treatment of pathological gambling]. PMID- 16691265 TI - [Written drug advertisements--are they reliable?]. AB - BACKGROUND: Drug advertisements affect the practice and continuous education of physicians. We assessed whether drug advertisements in Norway were in accordance with national regulations in the field. METHODS: All drug advertisements received by three general practitioners during a period of three months were collected. One advertisement for each of the 50 pharmaceutical products most frequently advertised was reviewed, available references obtained, and the information evaluated in relation to Norwegian regulations. RESULTS: A total of 294 advertisements for 77 different products were reviewed. The 50 chosen advertisements contained 191 reference citations, of which 93% were retrieved. The originator of the advertisement in question provided 72% of the references we asked for. We identified 262 promotional claims for which the regulations require a reference citation. Of these, 135 (52%) did not comply with the regulations, including 15 (6%) that were false. CONCLUSION: Only half of the information presented in drug advertisements was correct and clinically relevant. Relatively few statements were false, but a considerable proportion of statements gave an excessively positive picture of the product; hence, in general, this kind of information has no value as a source of information. PMID- 16691266 TI - [General practitioners' experiences with refugee patients]. AB - BACKGROUND: Refugees are likely to suffer from complex medical conditions due to persecution and exile. The symptoms may persist several years after the individual has fled his or her homeland. Knowledge of refugees' health condition in their receptive countries is insufficient. MATERIAL AND METHOD: Qualitative interviews with 12 general practitioners (GPs) in and around Oslo. A narrative approach was chosen for the analysis. RESULTS: The GPs' consultations with refugee patients seem to be characterised by uncertainty on the relevance of their refugee experience to their health condition. The GPs were reluctant to invite the patient to talk about traumatic experiences connected to their homelands, the escape and the exile. Rather, they express that unfamiliar culture and foreign language act as barriers to good communication between doctor and patient. Specialised health services seem neither to be accessible to the patients nor to be providers of clinical supervision for the GPs. INTERPRETATION: Cultural status seems to cover up experiences related to escape and exile. A lack of specialised health services seems to place the GP as a single actor in the medical service to refugee patients. The interviews revealed a need for instruction material adapted to the GPs' situation. PMID- 16691267 TI - [Pharmacological treatment of pathological gambling]. AB - BACKGROUND: The need for effective treatment for pathological gambling is urgent. The majority of treatment studies and available treatments today are based upon cognitive-behavioural therapy. Recently, however, several studies investigating the effects of pharmacological interventions have been published. We conducted a review of these studies. MATERIAL AND METHODS: Studies for inclusion were identified through searches in PubMed covering the period 1950 to June 2005. A total of 12 studies were included. RESULTS: Selective serotonin reuptake inhibitors and similar compounds were associated with improvement in two out of five placebo-controlled studies and in two out of three studies with a pre-post design. Opioid antagonists were associated with improvement in one placebo controlled study and in one study with a pre-post design. Mood stabilisers gave improvement in the one placebo-controlled study as well as in both studies with a pre-post design. INTERPRETATION: Pharmacotherapy may yield beneficial effects in the treatment of pathological gamblers. Still, more well-controlled studies with control of comorbid psychiatric conditions are needed. PMID- 16691268 TI - [The withdrawal of Vioxx: how did the patients react?]. AB - BACKGROUND: The withdrawal of rofecoxib (Vioxx) from the global market because of adverse cardiovascular effects affected many patients and created a lot of publicity. MATERIAL AND METHODS: 287 patients from four general practices who had received prescriptions for rofecoxib in 2004 were identified from the medical records of their general practitioner (GP) and mailed a questionnaire addressing their reactions and their expectations of their GP in this respect. RESULTS: 140 out of 287 patients returned the questionnaire. More than half were scared by the media frenzy about the withdrawal. 15% had received information from their GP after the withdrawal, whereas 70% wanted this kind of information. 42% had received prescriptions for rofecoxib, even if the patients could not confirm the presence of the diagnostic criteria. INTERPRETATION: The media frenzy gave rise to concern among patients, who want their GP to be more active in informing them about their use of unsafe medication. Better routines and data tools are needed for GPs to better identify and inform patients when needed. PMID- 16691269 TI - [Autotransfusion in prosthetic hip surgery]. AB - BACKGROUND: Homologous transfusion implies a risk for transmission of infectious diseases and transfusion reactions. The aim of the present study was to compare homologue transfusion with mechanical autotransfusion perioperatively in prosthetic hip surgery. METHODS: 111 patients were included in the study; group 1: 29 patients operated between June 2001 and June 2002 (before using autotransfusion); group 2: 35 patients operated between September 2002 and March 2003 (using common autotransfusion); group 3: 47 patients operated after March 2003 (using common autotransfusion and salvaging of rinsing fluid). RESULTS: Postoperative hemoglobin was significantly higher in group 3 (10,9 +/- 0.3 g/dl) than in group 2 (9,6 +/- 0.3 g/dl; p < 0.01) and group 1 (9,5 +/- 0,4 g/dl; p < 0.01). Amount of autologous erythrocyte concentrate in group 3 was significant higher than in group 2 (440 +/- 51 ml vs. 238 31 ml; p < 0.01). There were fewer patients in group 3 (9%) than in group 2 (29%; p < 0.04) and in group 1 (76%; < 0.01) who required homologous blood. CONCLUSION: The use of perioperative blood collection and retransfusion significantly reduces homologue blood transfusion in prosthetic hip surgery. PMID- 16691270 TI - [Surgical treatment of hormone-producing pituitary adenomas]. AB - BACKGROUND: Transsphenoidal surgical treatment is established as a treatment of hormonally active pituitary tumours, except prolactinomas. The cure rates in studies of large series of patients are varying. The volume of patients at Haukeland University Hospital, Bergen is limited, and in order to maintain sufficient experience, one surgeon performs all procedures. The aim of this study was to study the rates of cure and complications in patients who had undergone this procedure. METHODS: We did a retrospective survey evaluating the operative results of one surgeon operating hormonally active pituitary tumours from 1993 through 2002 at Haukeland University Hospital. Data were collected from medical and surgical case records. RESULTS: Primary surgery cured 37 out of 57 patients. There were 13 complications, including one death in hospital. Twenty patients received additional treatment with surgery, radiation therapy, or adrenalectomy. Thirteen of these patients received medical treatment with either somatostatin analogues or dopamine agonists. INTERPRETATION: Our study indicates that one surgeon can achieve results for transsphenoidal surgery that are on par with results reported from larger centres. PMID- 16691271 TI - [When should the trauma team be activated?]. AB - BACKGROUND: On admission to hospital, severely injured patients should be received by a trauma team so that the prospects of early life-saving treatment are not precluded. Considerations about use of resources indicate that the team is not activated when the likelihood of serious injury is low. A recommendation with criteria for activation of the trauma team has been established at the University Hospital of North Norway. We have studied how the recommendations were followed and calculated over- and under-triage and to highlight the question of whether activation should rather be automatically implemented based on predefined criteria. MATERIAL AND METHODS: We use descriptive statistics to analyse compliance with the recommendation as well as pre- and intra-hospital data for trauma patients who prior to admission were recognised as having fulfilled the criteria. RESULTS: Of the 109 trauma victims who, according to the recommendation, from 1 June 2001 to 31 May 2002 should have been received by the trauma team, 59 were received by a surgeon alone. Given that all patients with an injury severity score of 16 or more should be received by a trauma team, under utilisation of the team was 50%, whereas the proportion of excess calls was 58%. Elimination of under-utilisation among this group of patients would, by strict adherence to the criteria, have conferred a very modest increase in the proportion of superfluous calls to 61%. INTERPRETATION: The trauma team should be activated automatically in accordance with predefined criteria. PMID- 16691272 TI - [Contact allergy in the mouth]. AB - Contact allergy is not frequently seen in the oral mucosa. Oral lichenoid reactions in conjunction with dental restorations, particularly amalgam fillings, are among the most common affections. Subjective symptoms such as burning mouth syndrome, could have an allergic component, but the etiology is unclear and complex. Food additives and oral hygiene products can cause oral lesions of allergic nature. Patch testing is the routine diagnostic tool used by dermatologists, though the interpretation of the clinical relevance is crucial. PMID- 16691273 TI - [Adverse drug effects in the oral region]. AB - Syphilis was previously termed "the great imitator" because so many of the signs and symptoms are indistinguishable from those of other diseases. This role has been taken over by drugs, and this also applies to adverse drug reactions in the oral region. Accordingly, a careful drug history, including identification of any prescription, over-the-counter, or herbal medicines used, may give an important clue to the differential diagnosis of oral diseases when the aetiology is not apparent. Virtually all drugs have the potential to cause oral adverse reactions, but some have a greater ability to do so than others. Among the numerous adverse oral manifestations are xerostomia, taste disturbances and ulceration. The reactions are often non-specific, but they may mimic specific disease states such as erythema multiforme, lichen planus and pemphigus. Drug-induced gingival hyperplasia is an example of a quite characteristic and easily recognisable oral side effect. This article briefly describes some of the presentations and mechanisms of oral manifestations of drug therapy and the drugs that most commonly are responsible. Just like approved pharmaceuticals, herbal medicines are also associated with adverse oral manifestations. Finally we comment on some of the more recent reports on osteonecrosis of the jaws associated with the use of bisphosphonates. PMID- 16691274 TI - [How is the oral cavity affected by cancer treatment?]. PMID- 16691275 TI - [Should specialist education in general medicine be more effective?]. PMID- 16691279 TI - [Hemodynamic renal changes in right-sided heart failure]. PMID- 16691282 TI - [Good practice--what is it?]. PMID- 16691288 TI - Infrared spectroscopic characterization of mineralized tissues. AB - Vibrational spectroscopy (Infrared and Raman), and in particular micro spectroscopy and micro-spectroscopic imaging has been used to characterize developmental changes in bone and other mineralized tissues, to monitor these changes in cell cultures, and to detect disease and drug-induced modifications. Examples of the use of infrared micro-spectroscopy and micro-spectroscopic imaging are discussed in this review. PMID- 16691290 TI - Group membership and everyday social comparison experiences. AB - In two everyday experience studies, we examined the degree to which everyday social comparisons are framed by group membership. In the first study, 30 undergraduates attending a public university in the United States completed short questionnaires about their social comparison experiences whenever they were signalled. In the second study, 34 ethnic minority undergraduates from the same university completed similar questionnaires about their social comparison experiences. Across both studies, comparisons in which participants viewed themselves as an ingroup member in comparison to an outgroup comprised less than 10% of the comparison experiences reported by participants. However, minorities in the second study who reported closer identification with their ethnic group reported more comparison experiences in which they mentioned their own or the comparison target's ethnicity. PMID- 16691291 TI - MCP-1 contributes to macrophage infiltration into adipose tissue, insulin resistance, and hepatic steatosis in obesity. AB - Adipocytes secrete a variety of bioactive molecules that affect the insulin sensitivity of other tissues. We now show that the abundance of monocyte chemoattractant protein-1 (MCP-1) mRNA in adipose tissue and the plasma concentration of MCP-1 were increased both in genetically obese diabetic (db/db) mice and in WT mice with obesity induced by a high-fat diet. Mice engineered to express an MCP-1 transgene in adipose tissue under the control of the aP2 gene promoter exhibited insulin resistance, macrophage infiltration into adipose tissue, and increased hepatic triglyceride content. Furthermore, insulin resistance, hepatic steatosis, and macrophage accumulation in adipose tissue induced by a high-fat diet were reduced extensively in MCP-1 homozygous KO mice compared with WT animals. Finally, acute expression of a dominant-negative mutant of MCP-1 ameliorated insulin resistance in db/db mice and in WT mice fed a high fat diet. These findings suggest that an increase in MCP-1 expression in adipose tissue contributes to the macrophage infiltration into this tissue, insulin resistance, and hepatic steatosis associated with obesity in mice. PMID- 16691292 TI - Neuronal calcium sensor-1 enhancement of InsP3 receptor activity is inhibited by therapeutic levels of lithium. AB - Regulation and dysregulation of intracellular calcium (Ca2+) signaling via the inositol 1,4,5-trisphosphate receptor (InsP3R) has been linked to many cellular processes and pathological conditions. In the present study, addition of neuronal calcium sensor-1 (NCS-1), a high-affinity, low-capacity, calcium-binding protein, to purified InsP3R type 1 (InsP3R1) increased the channel activity in both a calcium-dependent and -independent manner. In intact cells, enhanced expression of NCS-1 resulted in increased intracellular calcium release upon stimulation of the phosphoinositide signaling pathway. To determine whether InsP3R1/NCS-1 interaction could be functionally relevant in bipolar disorders, conditions in which NCS-1 is highly expressed, we tested the effect of lithium, a salt widely used for treatment of bipolar disorders. Lithium inhibited the enhancing effect of NCS-1 on InsP3R1 function, suggesting that InsP3R1/NCS-1 interaction is an essential component of the pathomechanism of bipolar disorder. PMID- 16691293 TI - Steroid and xenobiotic receptor and vitamin D receptor crosstalk mediates CYP24 expression and drug-induced osteomalacia. AB - The balance between bioactivation and degradation of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] is critical for ensuring appropriate biological effects of vitamin D. Cytochrome P450, family 24-mediated (CYP24-mediated) 24-hydroxylation of 1,25(OH)2D3 is an important step in the catabolism of 1,25(OH)2D3. The enzyme is directly regulated by vitamin D receptor (VDR), and it is expressed mainly in the kidney, where VDR is also abundant. A recent report suggests that activation of steroid and xenobiotic receptor (SXR) also enhances the expression of CYP24, providing a new molecular mechanism of drug-induced osteomalacia. However, here we showed that activation of SXR did not induce CYP24 expression in vitro and in vivo, nor did it transactivate the CYP24 promoter. Instead, SXR inhibited VDR mediated CYP24 promoter activity, and CYP24 expression was very low in tissues containing high levels of SXR, including the small intestine. Moreover, 1,25(OH)2D3-induced CYP24 expression was enhanced in mice lacking the SXR ortholog pregnane X receptor, and treatment of humans with the SXR agonist rifampicin had no effect on intestinal CYP24 expression, despite demonstration of marked CYP3A4 induction. Combined with our previous findings that CYP3A4, not CYP24, plays the dominant role in hydroxylation of 1,25(OH)2D3 in human liver and intestine, our results indicate that SXR has a dual role in mediating vitamin D catabolism and drug-induced osteomalacia. PMID- 16691294 TI - IL-15 induces CD4 effector memory T cell production and tissue emigration in nonhuman primates. AB - HIV infection selectively targets CD4+ effector memory T (T EM) cells, resulting in dramatic depletion of CD4+ T cells in mucosal effector sites in early infection. Regeneration of the T EM cell compartment is slow and incomplete, even when viral replication is controlled by antiretroviral therapy (ART). Here, we demonstrate that IL-15 dramatically increases in vivo proliferation of rhesus macaque (RM) CD4+ and CD8+ T EM cells with little effect on the naive or central memory T (T CM) cell subsets, a response pattern that is quite distinct from that of either IL-2 or IL-7. T EM cells produced in response to IL-15 did not accumulate in blood. Rather, 5-bromo-2'-deoxyuridine (BrdU) labeling studies suggest that many of these cells rapidly disperse to extralymphoid effector sites, where they manifest (slow) decay kinetics indistinguishable from that of untreated controls. In RMs with uncontrolled SIV infection and highly activated immune systems, IL-15 did not significantly increase CD4+ T EM cell proliferation, but with virologic control and concomitant reduction in immune activation by ART, IL-15 responsiveness was again observed. These data suggest that therapeutic use of IL-15 in the setting of ART might facilitate specific restoration of the CD4 + T cell compartment that is the primary target of HIV with less risk of exhausting precursor T cell compartments or generating potentially deleterious regulatory subsets. PMID- 16691296 TI - "Unintentional" acetaminophen overdose on the rise: who is responsible? Dr Robert J Fontana is interviewed by Paul C Adams. PMID- 16691295 TI - Salt-sensitive hypertension is associated with dysfunctional Cyp4a10 gene and kidney epithelial sodium channel. AB - Functional and biochemical data have suggested a role for the cytochrome P450 arachidonate monooxygenases in the pathophysiology of hypertension, a leading cause of cardiovascular, cerebral, and renal morbidity and mortality. We show here that disruption of the murine cytochrome P450, family 4, subfamily a, polypeptide 10 (Cyp4a10) gene causes a type of hypertension that is, like most human hypertension, dietary salt sensitive. Cyp4a10-/- mice fed low-salt diets were normotensive but became hypertensive when fed normal or high-salt diets. Hypertensive Cyp4a10-/- mice had a dysfunctional kidney epithelial sodium channel and became normotensive when administered amiloride, a selective inhibitor of this sodium channel. These studies (a) establish a physiological role for the arachidonate monooxygenases in renal sodium reabsorption and blood pressure regulation, (b) demonstrate that a dysfunctional Cyp4a10 gene causes alterations in the gating activity of the kidney epithelial sodium channel, and (c) identify a conceptually novel approach for studies of the molecular basis of human hypertension. It is expected that these results could lead to new strategies for the early diagnosis and clinical management of this devastating disease. PMID- 16691297 TI - Hilar strictures - clear management strategies start to emerge. PMID- 16691298 TI - Nitric oxide and prostaglandins potentiate the liver regeneration cascade. AB - The liver has the remarkable ability to regenerate following damage or surgical resection. Although this feature of the liver has been studied for over 100 years, the trigger of the liver regeneration cascade remains controversial. Recent experimental evidence supports the hypothesis that nitric oxide (NO) and prostaglandins (PGs), released secondary to an increase in the blood flow-to liver mass ratio following two-thirds partial hepatectomy (PHx), work synergistically to trigger liver regeneration. To extend this research, the hypothesis that NO and PGs are potential therapeutic targets to potentiate the liver regeneration cascade is tested. The NO donor s-nitroso-n acetylpenicillamine, the phosphodiesterase V antagonist zaprinast (ZAP) and PGI2 each potentiated c-fos messenger RNA expression, an index of initiation of the liver regeneration cascade, following PHx. Also, the triple combination of s nitroso-n-acetylpenicillamine, ZAP and PGI2 potentiated c-fos messenger RNA expression. These results support the hypothesis that NO and PGs can potentiate initiation of the regeneration cascade. An additional index of liver weight restoration 48 h after PHx was also used to test the hypothesis, because this index encompasses the entire liver regeneration cascade. ZAP and 6-keto PGF1alpha, a stable metabolite of PGI2, and the combination of ZAP and 6-keto PGF1alpha, each potentiated liver weight restoration 48 h after PHx. These results also provide support for the hypothesis that NO and PGs are possible therapeutic targets to potentiate liver regeneration following surgical resection. PMID- 16691299 TI - Clinical diagnosis of achalasia: how reliable is the barium x-ray? AB - Manometry is considered to be the gold standard for the diagnosis of achalasia. However, many physicians believe that contrast radiography, classically showing esophageal dilation with bird-beak narrowing of the gastroesophageal junction, is also accurate in either diagnosing or excluding the disorder. The aim of the current study was to determine the accuracy of barium x-ray in the diagnosis of achalasia. The radiological diagnosis of all patients manometrically diagnosed with achalasia (using conventional criteria) between January 1994 and June 1998 were reviewed. A total of 51 cases of achalasia were identified. Thirteen patients were excluded because they either did not have contrast radiography before a manometric diagnosis or had their x-rays performed more than six months previously. Of the remaining 38 patients, achalasia was stated as a diagnostic possibility in the radiologists report in only 22 (58%) of those patients. Achalasia was not considered in the remaining 16 patients: two were reported as normal, four as having stenosis/narrowing in distal esophagus, two as having presbyesophagus, one as having mild gastroesophageal reflux and seven as having nonspecific dysmotility. To determine the reason for the diagnostic failure of the barium x-ray, an expert gastrointestinal radiologist reviewed 12 of the nondiagnostic x-rays in a blinded fashion, interspersed with 10 randomly selected esophageal-contrast radiographs from control subjects to avoid bias. Of these initially nondiagnostic x-rays in achalasia patients, typical radiological features of achalasia were deemed to be present in 50%. The present study indicates that contrast radiography lacks sensitivity in the diagnosis of achalasia. This is not only due to radiologist oversight but also because of the absence of the characteristic radiological features in many cases. This reinforces the important role of esophageal manometry in patients with persistent nonstructural dysphagia. PMID- 16691300 TI - Adult-to-adult living donor liver transplantation. AB - The present review outlines the principles of living donor liver transplantation, donor workup, procedure and outcomes. Living donation offers a solution to the growing gap between the need for liver transplants and the limited availability of deceased donor organs. With a multidisciplinary team focused on donor safety and experienced surgeons capable of performing complex resection/reconstruction procedures, donor morbidity is low and recipient outcomes are comparable with results of deceased donor transplantation. PMID- 16691301 TI - Non alcoholic fatty liver disease: a clinical approach and review. AB - Nonalcoholic fatty liver disease (NAFLD) is the most common cause of incidental elevation of liver enzymes in North America and Europe. Risk factors for NAFLD include body mass index of 25 kg/m2 or greater, central obesity and diabetes mellitus. The spectrum of disease is variable, ranging from simple steatosis with benign prognosis, to non-alcoholic steatohepatitis and cirrhosis, conferring increase in morbidity and mortality. The primary abnormality or 'first hit' in patients with NAFLD is insulin resistance leading to hepatic steatosis. The second hit involves multiple proinflammatory cytokines resulting in non-alcoholic steatohepatitis. Treatment is aimed at aggressive risk factor control and weight loss. Currently, there are no pharmacological agents recommended in the treatment of NAFLD, although preliminary studies suggest promising agents in the future. PMID- 16691302 TI - Rofecoxib-induced hepatotoxicity: a forgotten complication of the coxibs. AB - Rofecoxib is a member of the coxib family of nonsteroidal anti-inflammatory drugs that selectively inhibit cyclooxygenase-2. Although the coxibs are generally well tolerated, rofecoxib was recently withdrawn from the market due to concerns regarding cardiovascular safety. Rare cases of hepatic injury attributable to the coxibs have been reported. In the present study, two additional cases of severe hepatotoxicity are described in patients with cholestatic symptoms and abnormal liver biochemistry, shortly following the initiation of rofecoxib for arthritic complaints. In both cases, liver histology was compatible with drug-induced hepatotoxicity, and rapid clinical and biochemical improvements were observed following rofecoxib discontinuation. With new coxibs and expanding indications on the horizon, physicians in all areas of practice must be aware of this disorder and consider it in any patient who develops hepatic dysfunction after taking a coxib. PMID- 16691303 TI - Resolution of paraneoplastic collagenous enterocolitis after resection of colon cancer. AB - A 52-year-old woman developed severe watery diarrhea, weight loss, anemia and hypoalbuminemia. A localized colon cancer was detected. Subsequently, extensive collagenous mucosal involvement of the small and large intestine was discovered. After resection of the colon cancer, her symptoms resolved. In addition, resolution of the inflammatory process occurred, including the subepithelial collagen deposits. Despite extensive small and large intestinal involvement, both clinical and histological resolution of collagenous inflammatory disease was evident. Collagenous enterocolitis is an inflammatory process that may represent a distinctive and reversible paraneoplastic phenomenon. PMID- 16691304 TI - Eosinophilic pancreatitis mimicking pancreatic neoplasia. AB - Eosinophilic pancreatitis (EP) is a rare disease. It typically occurs in the setting of either eosinophilic gastroenteritis or the hypereosinophilic syndrome. Isolated eosinophilic infiltration of the pancreas is less common. EP usually presents as a pancreatic tumour with abdominal pain and/or obstructive jaundice. The diagnosis is often not made until after pancreatic resection under suspicion of a pancreatic tumour. The case of a 14-year-old boy whose initial presentation was suggestive of a pancreatic tumour is reported. Radiological evaluation revealed a pancreatic mass suggestive of a pancreatic tumour obstructing the duodenum and common bile duct. The patient underwent surgery and a gastrojejunal anostomosis, tube cholecystostomy and biopsy were performed with no postoperative complications. The diagnosis of EP was established after surgical biopsy. The biopsy specimen revealed prominent eosinophilic infiltration. Serum immunoglobulin E levels were elevated. The patient was treated with oral prednisolone (40 mg/day). After two months of oral steroid therapy, clinical manifestations rapidly improved and peripheral eosinophilia subsided. Computed tomography scan revealed remission of the pancreatic mass-like lesion. PMID- 16691305 TI - Cryopreservation, encapsulation and promotion of shoot production of embryonic axes of a recalcitrant species Ekebergia capensis, Sparrm. AB - A study on zygotic axes of the recalcitrant seeds of Ekebergia capensis compared two cryopreservation methods, partial desiccation, and encapsulation-dehydration, and also investigated a method to promote shoot production. High (80 percent) survival (assessed as root production) was obtained after direct immersion into liquid nitrogen of axes rapidly dehydrated by flash drying for 20 min to a water content about 0.4 g water per g dry mass. In contrast, no survival at all was obtained of axes that were first encapsulated, then desiccated for three hours to the same water concentration as those fast-dried, and then placed in a cryovial and immersed in liquid nitrogen. Axes encapsulated after cryopreservation germinated both in vitro and in soil, and could be stored at room temperatures for several weeks while maintaining germinability, thus producing synseeds capable of distribution. However, shoot production after cryopreservation was seldom observed. The inclusion of the plant growth regulator, N6-benzyl adenine (BA) in the MS-based recovery medium promoted vigorous multiple shoot formation. Microscopical examination of embryos of E. capensis revealed that the cotyledonary insertions were contiguous with the shoot apex, leading to the conclusion that injury to, and ultimate necrosis of, the apical meristem following severing of these connections was a primary cause of the observed lack of, or poor, shoot development in excised axes (whether cryopreserved or not). The study demonstrated that it may be possible to resolve two of the problems facing attempts at cryopreservation of axes of recalcitrant seeds; lack of shoot production and difficulty of distribution of cryopreserved material for re introduction. PMID- 16691306 TI - Characterisation of cryopreserved cells freshly isolated from human bone marrow. AB - Osteoblast progenitor cells (OBPCs) isolated from bone marrow have the ability to differentiate into osteoblasts and thus potential therapeutic use to tissue engineer bone. In order for OBPCs to be available for clinical use a means of storing viable cells is necessary. The aim of this study was to determine whether a simple method of cryopreservation had an effect on osteogenic differentiation or growth of OBPCs isolated from fresh human bone marrow. Stro-1 was used to identify the isolated OBPCs. The osteoblastic potential of the marrow cells was confirmed as culture with osteogenic supplements (OS) significantly increased osteoblastic protein production (alkaline phosphatase (ALP), osteopontin and osteocalcin) compared with standard conditions (P less than 0.05). Ten further marrow aspirates were harvested; each was halved for either cryopreservation or control culture. Primary cultures from both populations formed colonies with recognised OBPC morphology. OS stimulated both cryopreserved and control populations to produce significantly more osteoblastic proteins (P less than 0.05) and there was no significant difference between the increase in osteogenic proteins when cultured with OS (P great than 0.2). The proliferation rate after 5 days in culture was not significantly affected by cryopreservation (P greater than 0.05). It has been suggested that OBPCs are immuno-privileged; so allogenic cells could be implanted into patients for tissue engineering bone without causing a hypersensitivity reaction. Our study demonstrates a method of storage, which allows OBPCs to be available for use without affecting osteoblastic potential or viability. PMID- 16691307 TI - Cryopreservation of shoot tips of blackberry and raspberry by encapsulation dehydration and vitrification. AB - Encapsulation-dehydration and PVS2-vitrification cryopreservation protocols were evaluated for the long-term conservation of a diverse group of Rubus germplasm. Cold acclimation for a 4-week period prior to cryopreservation was necessary for regrowth of shoot apices from blackberry and raspberry genotypes. For the encapsulation-dehydration protocol, encapsulated apices were pretreated in 0.75 M sucrose for 20 h, desiccated 6-h under laminar flow to c. 20 percent moisture content, then plunged in liquid nitrogen (LN) and rapidly warmed. The PVS2 vitrification protocol included pretreating shoot tips on 5 percent dimethyl sulfoxide (DMSO) medium for 48 h, exposure to loading solution (LS) and PVS2 for 20 min each at 25 degree C , followed by immersion in LN and rapid warming. Shoot tips of 25 genotypes in 9 Rubus species and 9 Rubus hybrids were successfully cryopreserved with recovery of 60 to 100 percent using the encapsulation dehydration protocol. Four genotypes of 3 species were tested using the vitrification protocol with 71 percent average regrowth. The present results indicate that both of these improved cryopreservation protocols can be applied to a diverse range of Rubus genetic resources. PMID- 16691308 TI - Freeze-drying of human platelets: influence of intracellular trehalose and extracellular protectants. AB - Freeze-drying is an ideal alternative for long-term preservation of platelets in blood banks. Intracellular trehalose is believed to be an effective lyoprotectant for preserving cells during freeze-drying. In this study, 13 mM intracellular trehalose was loaded into human platelets through fluid-phase endocytosis pathway. Bovine serum albumin and trehalose were used as extracellular protectants. The effects of intracellular trehalose and extracellular protectants on freeze-dried platelets were studied respectively. The results showed 13 mM intracellular trehalose was beneficial to freeze-dried human platelets, but only slightly enhanced the protection afforded by extracellular protectants. Loaded with 13 mM intracellular trehalose, platelets were freeze-dried in a formulation of 1 percent bovine serum albumin and 1 percent trehalose, 40 days later, the survival rate of rehydrated platelets was about 85 percent, the morphology of rehydrated platelets was intact and the aggregation percentage with thrombin (1 U/ml) was 97.3 percent. PMID- 16691309 TI - Characteristics of fresh gander semen and its susceptibility to cryopreservation in six generations derived from geese inseminated with frozen-thawed semen. AB - The paper summarises seven years experiments designed to determine the effect of continuous insemination with frozen-thawed semen on fresh semen quality and sperm susceptibility to freezing stress in succeeding generations. During course of experiments, semen was collected from 10-12 White Koluda ganders at the age of 8 9 months, then subjected to freezing and used after thawing for insemination of 10 geese in order to obtain the subsequent generation of males. Semen was diluted 1 to 0.5 (v/v) with EK diluent, equilibrated for 15 min at +4 degrees C, mixed with 6 percent (v/v) of dimethyl-formamide (DMF), frozen to temp. -140 degrees C at a rate 60 degree C per min and then transferred into liquid nitrogen container. Semen samples were thawed prior to insemination in a 60 degree C water bath. It is difficult to conclude whether freezing stress affected the fresh semen quality, since average volume of SQF (index comprising ejaculate volume, sperm concentration and percentage of live normal cells) varied between generations from 19.3 to 56.2. Continuous goose reproduction by insemination with frozen-thawed semen resulted in significant increase (P less than 0.01) in spermatozoa resistance to cryoinjury in every subsequent generation. In the relation to adequate fresh semen the percentage of live morphologically intact spermatozoa which withstood freezing procedure increased from 27.2 in first generation to 74.4 in sixth generation. PMID- 16691310 TI - Ultra rapid freezing and vitrification of human embryos derived from abnormally fertilised zygotes. AB - The present study was undertaken to compare the developmental capacity of human embryos derived from abnormally fertilised zygotes (1 PN, > 3 PN; 16-18 hours after ICSI) cryopreserved using two techniques: ultra rapid freezing and vitrification. At 2-4 cell stage, (48 hours after ICSI), these abnormally fertilised embryos were then distributed in three groups: a) embryos that were cryopreserved by ultra rapid freezing (URF Group), b) embryos cryopreserved by vitrification (V Group) and c) embryos that were not cryopreserved (Control group). Survival rates and embryo development after 24 hours of in vitro culture (72 hours after ICSI) were compared. 42 embryos were cryopreserved by ultra rapid freezing in 0.5 mL straws, using a mixture of dimethyl sulphoxide (3M) and sucrose (0.25M) in a base solution consisting of IVF medium plus 20 percent (v/v) of Human Serum Albumin (HSA), and 24 embryos were vitrified in 0.25 ml straws, using a two step protocol with an equilibration solution consisting of 10 percent ethylene glycol (1.79 M) and 10 percent dimethyl sulphoxide (1.41 M) in a base solution of modified phosphate buffered saline (PBS) with 20 percent of HSA and a vitrification solution consisting of 20 percent ethylene glycol (3.58 M), 20 percent dimethyl sulphoxide (2.82 M) and 0.5 M sucrose in base solution. The recovery rate after thawing/warming was lower for the vitrification group (75 percent V; 83 percent URF). The number of embryos with less than 50 percent of intact blastomeres after cryopreservation was significantly higher for the URF group (0 percent V; 34 percent URF). After in vitro culture, the rate of embryos not cryopreserved (Control group) that developed in vitro (72 hours after ICSI) was the highest (86 percent), followed by group V (50 percent), while group URF was the lowest (13 percent). These differences were statistically significant. This straw method of vitrification is successful and safe. PMID- 16691311 TI - Incisional hernia after a tension-free vaginal tape procedure. AB - A case is presented of an incisional hernia of the inguinal canal presenting 9 months after a tension-free vaginal tape (TVT) procedure and anterior vaginal repair. The TVT and repair procedure was complicated by prolonged postoperative urinary retention requiring midline incision of the tape for resumption of normal voiding. The patient had a hysterectomy several years earlier via a Pfannenstiel incision. No other risk factors for hernia were identified. There are no previous reports of TVT-related incisional hernia. We conclude that incisional hernia is a rare complication of the TVT procedure and that the characteristics of the TVT tape may contribute to late occurrence of herniation. PMID- 16691312 TI - Role of the cyclooxygenase pathway in the protection against postischemic stunning in conscious sheep. AB - OBJECTIVE: There are controversial reports in conscious animals regarding the role of cyclooxygenase-2 in late preconditioning (LP). This study analyzed the effect of COX-2 involvement in non-preconditioned hearts (NP) and in mediation of LP protection against stunning in conscious sheep submitted to a prolonged reversible ischemia. METHODS: Six groups were considered: NP: 12 min ischemia and 120 min reperfusion; LP consisting of six periods of 5 min-ischemia-5 min reperfusion 24 h before the 12 min ischemia; NP and LP with either the non selective COX-1 and COX-2 inhibitor, aspirin (20 mg/kg), or the specific COX-2 inhibitor, celecoxib (3 mg/kg) before the 12 min ischemic period. RESULTS: Mean postischemic wall thickening fraction (as % of preischemic values) improved from 49.6 +/- 4.0% in NP to 72.5 +/- 3.5% in LP (p < 0.01) and a similar protection was obtained with aspirin and celecoxib in NP hearts (p < 0.01). Neither aspirin nor celecoxib administration prior to the prolonged ischemia on day 2 abrogated LP improvement of postischemic dysfunction. Moreover, LP with aspirin improved the protective response (80.7 +/- 2.6%) over that obtained with aspirin in NP hearts (66.6 +/- 4.7%, p < 0.05). This effect was not obtained with celecoxib. CONCLUSIONS: Aspirin and celecoxib showed that COX-2 has a detrimental effect on mechanical cardioprotection in NP hearts of conscious sheep submitted to a prolonged reversible ischemia, and does not seem to participate as mediator of LP. Aspirin revealed a similar COX-1 deleterious action, since only when both COX 1 and COX-2 were inhibited, LP was put in evidence adding functional improvement over that obtained in NP hearts treated with aspirin. PMID- 16691313 TI - ALR and liver regeneration. AB - Liver possesses the capacity to restore its tissue mass and attain optimal volume in response to physical, infectious and toxic injury. The extraordinary ability of liver to regenerate is the effect of cross-talk between growth factors, cytokines, matrix components and many other factors. In this review we present recent findings and existing information about mechanisms that regulate liver growth, paying attention to augmenter of liver regeneration. PMID- 16691314 TI - Curcumin combats against cigarette smoke and ethanol-induced lipid alterations in rat lung and liver. AB - BACKGROUND: Human population, in spite of the medical and scientific achievements, still fall as a prey to the evils of habitual smoking and alcohol, thus necessitating safer counteracting measures. OBJECTIVE: To evaluate the effect of cotreatment of curcumin (Curcuma longa) in rats subjected to acute exposure to cigarette smoke (CS) and ethanol (EtOH). METHODOLOGY: Of the four groups of experimental rats, a set of rats was subjected to whole body exposure to cigarette smoke along with ethanol administration serving as a model of CS+EtOH injury. Curcumin treatment was given to two sets of rats: (i) one set receiving simultaneous CS+EtOH and (ii) one set of normal rats without any administration. The other group of rats served as control. Blood, liver and lung of rats were selected for assessment of CS+EtOH injury as well as curcumin treatment. RESULT: Altered lipid, lipoprotein profile and bile acid excretion were observed in CS+EtOH rats along with premalignant pathological state in tissues. In treated rats, the levels were maintained at near-normal levels along with near-normal histology. CONCLUSION: This biochemical picture on cotreatment with curcumin suggests that curcumin could counteract the injurious effects of combined CS and EtOH and thus might help to reduce the risk of hyperlipidemic disorders which develop due to smoking and drinking. PMID- 16691315 TI - Garlic compound, diallyl disulfide induces cell cycle arrest in prostate cancer cell line PC-3. AB - Prostate cancer is the most predominant cancer in men and related death rate increases every year. Till date, there is no effective therapy for androgen independent prostate cancer. Previous studies reported that aged garlic extract suppresses cancer growth. In the present study, diallyl disulfide [DADS], oil soluble organosulfur compound of garlic, was studied for its antiproliferative and induction of cell cycle arrest on prostate cancer cells in vitro. The suppression of cell growth was assessed by MTT assay. Induction of cell cycle arrest was assessed and confirmed by propidium iodide staining in flowcytometric analysis and western blotting analysis of major cell cycle regulator proteins. The results showed that DADS inhibited the growth of prostate cancer cells in a dose dependent manner, compared to the control. At 25 microM and 40 microM concentrations, DADS induced cell cycle arrest at G2/M transition in PC-3 cells. Western blotting analysis of cyclin A, B(1) and cyclin dependent kinase 1 [CDK1] revealed that DADS inhibited the cell cycle by downregulating CDK1 expression. It is concluded that DADS, inhibits proliferation of prostate cancer cells through cell cycle arrest. Dose dependent effect of DADS on PC-3 cell line was observed in the present study. PMID- 16691316 TI - Therapeutic potential of riboflavin, niacin and ascorbic acid on carbohydrate metabolizing enzymes in secondary endometrial carcinoma bearing rats. AB - Curative potential of riboflavin, niacin and ascorbic acid against tamoxifen mediated endometrial carcinoma was established by studies on carbohydrate metabolizing enzymes. The enzymes investigated were glycolytic enzymes namely, hexokinase; aldolase; phosphoglucoisomerase and the gluconeogenic enzymes namely, glucose-6-phosphatase and fructose-1, 6-biphosphatase in endometrial carcinoma bearing rats. A significant increase in glycolytic enzymes and a subsequent decrease in gluconeogenic enzymes were observed in plasma, liver and kidney of endometrial carcinoma animals. The administration of riboflavin (45 mg/kg bw/day), niacin (100 mg/kg bw/day) and ascorbic acid (200 mg/kg bw/day) along with tamoxifen (45 mg/kg bw/day) caused a significant decrease in the activity of glycolytic enzymes and a significant increase in the activities of gluconeogenic enzymes to near normal levels in experimental animals. Our results suggest that riboflavin, niacin and ascorbic acid have potential combination therapy against tamoxifen mediated secondary endometrial carcinoma in experimental rats. However, there were no deleterious side effects observed in combinants alone treated animals. PMID- 16691317 TI - The phosphoenolpyruvate carboxykinase of Mycobacterium tuberculosis induces strong cell-mediated immune responses in mice. AB - Phosphoenolpyruvate carboxykinase (PEPCK) catalyzes guanosine or adenosine mononucleotide-dependent reversible conversion of oxaloacetate (OAA) and phosphoenolpyruvate (PEP). Mycobacterium (M) tuberculosis possesses a putative GTP-dependent PEPCK. To analyze the immune responses caused by PEPCK, the effects of PEPCK on the induction of CD4(+) T cells and cytokines such as IFN-gamma, IL 12 and TNF-alpha were evaluated in mice. It was found that the number of CD4(+) T cells was increased in the PEPCK immunized mice although the change of the number of CD8(+) T cells was not significant. The cytokines IFN-gamma, IL-12 and TNF alpha were increased significantly in the mice immunized with PEPCK than those of incomplete adjuvant. These characteristics were further demonstrated in the mice infected by pckA mutated BCG strain. The results indicate that PEPCK can effectively induce cell-mediated immune response by increasing activity of cytokines and PEPCK may be a promising new subunit vaccine candidate for tuberculosis. PMID- 16691318 TI - Decreased bodyweight without rebound and regulated lipoprotein metabolism by gymnemate in genetic multifactor syndrome animal. AB - OBJECTIVE: The aim of this work was to find obesity control method without rebound. In our previous studies, gymnemate extracted from Gymnema sylvestre, inhibited oleic acid absorption. The Otsuka Long-Evans Tokushima Fatty (OLETF) rat, a genetic multifactor syndrome model, exhibits progressive overweight, hyperlipidemia and hyperglycemia. The effect of gymnemate on obesity in OLETF was investigated. METHODS: Three groups were divided (n=4-8): (1) OLETF-gymnemate, gymnema water extract (containing gymnemate) diet (62.5 g/kg) and water (2.5 g/kg) were supplied 2 weeks from 26-28 weeks, following it general diet and water were fed 3 weeks to observe if it rebound, (2) OLETF-control and (3) the counterpart Long-Evans Tokushima Otsuka rats as normal-control. RESULTS: With gymnemate treatment, the food and water intake were decreased about 1/3 and 2/3, along with body weight reduced 57.2+/- 6.4 and 75.5+/- 6.3 g during 1 and 2 weeks respectively. In the end of experiment (3 weeks after gymnemate withdrawal), the body weight was decreased to no significant difference with normal-control. The total cholesterol was decreased about 1/3, moreover LDL+VLDL (low-density and very-low-density lipoprotein) cholesterol decreased about 1/2. The proportion of HDL (high-density lipoprotein) cholesterol to the total cholesterol was increased. The serum triglyceride was decreased to the 1/4 of OLETF control. The level of serum cholesterol and triglyceride was no significant difference in gymnemate group with normal group. CONCLUSION: Supplementation with gymnemate promoted weight loss by its ability to reduce hyperlipidemia, which was no withdrawal rebound: an important discovery. Supplementation with gymnemate is a novel therapeutic tool for weight management, especially in multifactor syndrome. PMID- 16691319 TI - Sensitivity to Sn2+ of the yeast Saccharomyces cerevisiae depends on general energy metabolism, metal transport, anti-oxidative defences, and DNA repair. AB - Resistance to stannous chloride (SnCl(2)) of the yeast Saccharomyces cerevisiae is a product of several metabolic pathways of this unicellular eukaryote. Sensitivity testing of different null mutants of yeast to SnCl(2) revealed that DNA repair contributes to resistance, mainly via recombinational (Rad52p) and error-prone (Rev3p) steps. Independently, the membrane transporter Atr1p/Snq1p (facilitated transport) contributed significantly to Sn(2+)-resistance whereas absence of ABC export permease Snq2p did not enhance sensitivity. Sensitivity of the superoxide dismutase mutants sod1 and sod2 revealed the importance of these anti-oxidative defence enzymes against Sn(2+)-imposed DNA damage while a catalase deficient mutant (ctt1) showed wild type (WT) resistance. Lack of transcription factor Yap1, responsible for the oxidative stress response in yeast, led to 3 fold increase in Sn(2+)-sensitivity. While loss of mitochondrial DNA did not change the Sn(2+)-resistance phenotype in any yeast strain, cells with defect cytochrome c oxidase (CcO mutants) showed gradually enhanced sensitivities to Sn(2+) and different spontaneous mutation rates. Highest sensitivity to Sn(2+) was observed when yeast was in exponential growth phase under glucose repression. During diauxic shift (release from glucose repression) Sn(2+)-resistance increased several hundred-fold and fully respiring and resting cells were sensitive only at more than 1000-fold exposure dose, i.e. they survived better at 25 mM than exponentially growing cells at 25 microM Sn(2+). This phenomenon was observed not only in WT but also in already Sn(2+)-sensitive rad52 as well as in sod1, sod2 and CcO mutant strains. The impact of metabolic steps in contribution to Sn(2+)-resistance had the following ranking: Resting WT cells > membrane transporter Snq1p > superoxide dismutases > transcription factor Yap1p >or= DNA repair >> exponentially growing WT cells. PMID- 16691320 TI - Age-associated iron accumulation in bone: implications for postmenopausal osteoporosis and a new target for prevention and treatment by chelation. AB - Iron accumulation in tissues is believed to be a characteristic of aged humans and a risk factor for some chronic diseases. However, it is not known whether age associated iron accumulation is part of the pathogenesis of postmenopausal osteoporosis that affects approximately one out three women worldwide. Here, we confirmed that this accumulation of iron was associated with osteopenia in ovariectomized (OVX) rats (a model of peri- and postmenopausal osteoporosis due to estrogen deficiency). To further investigate whether the increased iron level plays a causal role in the onset of bone loss, we treated OVX rats with an orally active and bone targeted chelator that prevented iron accumulation in their skeletal tissues. The results showed that this treatment mitigated the loss of bone mass and the deterioration of bone micro-architecture. We also found that one possible mechanism of the protective action of iron chelation was to significantly reduce bone resorption. Thus, these findings provide a novel target and a potentially useful therapeutic strategy for the prevention and treatment of postmenopausal osteoporosis and perhaps other age-related diseases. PMID- 16691321 TI - A two-stage design for multiple testing in large-scale association studies. AB - Modern association studies often involve a large number of markers and hence may encounter the problem of testing multiple hypotheses. Traditional procedures are usually over-conservative and with low power to detect mild genetic effects. From the design perspective, we propose a two-stage selection procedure to address this concern. Our main principle is to reduce the total number of tests by removing clearly unassociated markers in the first-stage test. Next, conditional on the findings of the first stage, which uses a less stringent nominal level, a more conservative test is conducted in the second stage using the augmented data and the data from the first stage. Previous studies have suggested using independent samples to avoid inflated errors. However, we found that, after accounting for the dependence between these two samples, the true discovery rate increases substantially. In addition, the cost of genotyping can be greatly reduced via this approach. Results from a study of hypertriglyceridemia and simulations suggest the two-stage method has a higher overall true positive rate (TPR) with a controlled overall false positive rate (FPR) when compared with single-stage approaches. We also report the analytical form of its overall FPR, which may be useful in guiding study design to achieve a high TPR while retaining the desired FPR. PMID- 16691324 TI - Use of quantitative real-time PCR to investigate the dynamics of the red tide dinoflagellate Lingulodinium polyedrum. AB - A new method based on quantitative real-time polymerase chain reaction (qPCR) was developed and applied to quantify the red tide dinoflagellate Lingulodinium polyedrum in natural seawater samples and in laboratory cultures. The method uses a Molecular Beacontrade mark approach to target a species-specific region of the small subunit ribosomal RNA gene. The accuracy of the method was verified by microscopical counts using cultures of the dinoflagellate isolated from coastal waters near Los Angeles, CA, and with natural water samples spiked with cultured L. polyedrum. The method was applied to document the pattern and timing of vertical migration by the dinoflagellate in a 2-m water column on an 11:13 h light/dark photoperiod established in the laboratory. Positive phototaxis of L. polyedrum resulted in dense aggregations of the dinoflagellate within the top few centimeters of the water column during the light period. This pattern of distribution was readily established by both methods, although abundances of L. polyedrum determined using qPCR were higher than abundances determined by microscopy in the morning and lower in the afternoon and evening. These differences may have been a consequence of variability in the DNA content per cell because of synchrony of cell division. Counts using both methods to analyze natural samples collected from coastal waters in the Long Beach-Los Angeles area and adjacent San Pedro Channel were in close agreement. However, the qPCR method exhibited greater sensitivity than the microscopical method when L. polyedrum was present at low abundances, and qPCR had a much higher rate of sample throughput than microscopy. The development of this new approach for enumerating L. polyedrum provides a useful tool for studying the ecology of this important red tide species. PMID- 16691325 TI - Influence of hydrological pulse on bacterial growth and DOC uptake in a clear water Amazonian lake. AB - This study was conducted to evaluate: (1) the bacterial growth and the dissolved organic carbon (DOC) uptake in an Amazonian lake (Lake Batata) at high-water and low-water periods of the flood pulse; (2) the influence of nitrogen and phosphorus (NP) additions on bacterial growth and DOC uptake in Lake Batata at two flood pulse periods; and (3) the bioavailability of the main DOC sources in Lake Batata. Lake Batata is a typical clear-water Amazonian lake, located in the watershed of Trombetas River, Central Amazon, Brazil. Bacterial batch cultures were set up with 90% 0.2-microm filtered water and 10% inoculum from Lake Batata. N-NH(4)NO(3) and P-KH(2)PO(4), with final concentrations of 50 and 5 microM, respectively, were added to the cultures, except for controls. Extra sources of DOC (e.g., algal lysate, plant leachates) were added to constitute six distinct treatments. Bacterial response was measured by maximum bacterial abundance and rates of bacterial production, respiration, DOC uptake, and bacterial growth efficiency (BGE). Bacterial growth and DOC uptake were higher in NP treatments than in controls, indicating a consistent nutrient limitation in Lake Batata. The composition of DOC also seems to be an important regulating factor of bacterial growth in Lake Batata. Seasonally, bacterial growth and DOC bioavailability were higher at low-water period, when the phytoplankton is a significant extra source of DOC, than at high-water period, when the forest is the main source of DOC. DOC bioavailability was better estimated based on the diversity and the diagenetic stage of carbon compounds than on single classes of labile compounds. Changes in BGE were better related to CNP stoichiometry in the water, and the "excess" of organic substrates was oxidized in catabolism, despite the quality of these compounds for bacterial growth. Finally, we conclude that bacterial growth and DOC uptake vary throughout the flood pulse in clear-water Amazonian ecosystems as a result of changes in nutrient concentration and in DOC composition. PMID- 16691326 TI - Bacterioplankton growth and nutrient use efficiencies under variable organic carbon and inorganic phosphorus ratios. AB - We carried out enclosure experiments in an unproductive lake in northern Sweden and studied the effects of enrichment with different dissolved organic carbon (glucose)/inorganic phosphorous (DOC/Pi) ratios on bacterioplankton production (BP), growth efficiency (BGE), nutrient use efficiency (BNUE), growth rate, and specific respiration. We found considerable variation in BP, BGE, and BNUE along the tested DOC/Pi gradient. BGE varied between 0.87 and 0.24, with the highest values at low DOC/Pi ratios. BNUE varied between 40 and 9 g C g P(-1), with high values at high DOC/Pi ratios. More DOC was thus allocated to growth when bacteria tended to be C-limited, and to respiration when bacteria were P-limited. Specific respiration was positively correlated with bacterial growth rate throughout the gradient. It is therefore possible that respiration was used to support growth in P-limited bacteria. The results indicated that BP can be limited by Pi when BNUE is at its maximum, by organic C when BGE is at its maximum, and by dual organic C and Pi limitation when BNUE and BGE have suboptimal values. PMID- 16691327 TI - The community and phylogenetic diversity of biological soil crusts in the Colorado Plateau studied by molecular fingerprinting and intensive cultivation. AB - We studied the bacterial communities in biological soil crusts (BSCs) from the Colorado Plateau by enrichment and cultivation, and by statistically analyzed denaturing gradient gel electrophoresis (DGGE) fingerprinting of environmental 16S rRNA genes, and phylogenetic analyses. Three 500-m-long transects, tens of km apart, consisting of 10 equally spaced samples each, were analyzed. BSC communities consistently displayed less richness (10-32 detectable DGGE bands per sample) and Shannon diversity (2.1-3.3) than typical soil communities, with apparent dominance by few members. In spite of some degree of small-scale patchiness, significant differences in diversity and community structure among transects was detectable, probably related to the degree of crust successional maturity. Phylogenetic surveys indicated that the cyanobacterium Microcoleus vaginatus was dominant, with M. steenstrupii second among phototrophs. Among the 48 genera of nonphototrophs detected, Actinobacteria (particularly Streptomyces spp.) were very common and diverse, with 18 genera and an average contribution to the total 16S rDNA amplificate of 11.8%. beta-Proteobacteria and Bacteriodetes contributed around 10% each; Low-GC Gram-positives, alpha-Proteobacteria, Thermomicrobiales, and Acidobacteria were common (2-5%). However, the second largest contribution was made by deep-branching unaffiliated alleles (12.6%), with some of them representing candidate bacterial divisions. Many of the novel strains isolated are likely new taxa, and some were representatives of the phylotypes detected in the field. The mucoid or filamentous nature of many of these isolates speaks for their role in crust formation. PMID- 16691328 TI - Microbial colonization of beech and spruce litter--influence of decomposition site and plant litter species on the diversity of microbial community. AB - The present study was conducted to investigate the effect of decomposition site and plant litter species on the colonizing microbial communities. For this, litter bag technique using beech and spruce litter was combined with RNA-based fingerprinting and cloning. Litter bags were incubated for 2 and 8 weeks in the Ah horizon of beech and beech-spruce mixed forest sites. Although sugars and starch were rapidly lost, lignin content increased by more than 40% for beech and more than doubled for spruce litter at both soil sites at the end of the experiment. Denaturing gradient gel electrophoresis analysis of 16S and 18S rRNA RT-PCR products was used for screening of differences between bacterial and fungal communities colonizing the two litter types. Development of the microbial community over time was observed to be specific for each litter type and decomposition site. RT-PCR products from both litter types incubated in beech spruce mixed forest site were also cloned to identify the bacterial and fungal colonizers. The 16S rRNA clone libraries of beech litter were dominated by gamma proteobacterial members, whereas spruce libraries were mainly composed of alpha-, beta-, and gamma-proteobacterial members. Ascomycota members dominated the 18S rRNA clone libraries. Clones similar to Zygomycota were absent from spruce, whereas those similar to Basidiomycota and Glomeromycota were absent from beech libraries. Selective effects of litter quality were observed after 8 weeks. The study provides an insight into the bacterial and fungal communities colonizing beech and spruce litter, and the importance of litter quality and decomposition site as key factors in their development and succession. PMID- 16691329 TI - Society of American Gastrointestinal Endoscopic Surgeons (SAGES) guidelines on continuing medical education and financial relationships. PMID- 16691345 TI - The antibacterial effects of zinc ion migration from zinc-based glass polyalkenoate cements. AB - Zinc-based glass polyalkenoate cements have been synthesised and their potential use in orthopaedic applications investigated. Zinc ions were released from the materials in a rapid burst over the first 24 h after synthesis, with the release rate falling below detectable levels after 7 days. Cement-implanted bone samples were prepared and the released zinc was shown, using energy dispersive X-ray analysis, to penetrate from the cement into the adjacent bone by up to 40 microm. Finally, the cements exhibited antibacterial activity against Streptococcus mutans and Actinomyces viscosus that reflected the pattern of zinc release, with the inhibition of growth greatest shortly after cement synthesis and little or no inhibition measureable after 30 days. PMID- 16691346 TI - Genotoxicity and cytotoxicity of glass ionomer cements on Chinese hamster ovary (CHO) cells. AB - Glass ionomer cements are widely used in dentistry as restorative materials and adhesives for composite restorations. However, the results of genotoxicity studies using these materials are inconclusive in literature. The goal of this study was to examine the genotoxic and cytotoxic potential of three different glass ionomer cements available commercially (Ketac Cem, Ketac Molar and Vitrebond) by the single cell gel (comet) assay and trypan blue exclusion test, respectively. For this, such materials were exposed to Chinese hamster ovary (CHO) cells in vitro for 1 h at 37( composite function)C. Data were assessed by Kruskall-Wallis nonparametric test. The results showed that the powder from Ketac Molar displayed genotoxicity only in the maximum concentration evaluated (100 microg/mL). In the same way, the liquid from Vitrebond at 0.1% dilution caused an increase of DNA injury. Significant differences (P<0.05) in cytotoxicity provoked by all powders tested of glass ionomer cements were observed for exposure at 1,000 microg/mL concentration. With respect to liquids of glass ionomer cements evaluated, the major toxic effect on cell viability was produced at 10%, beginning at the dilution of 0.5% for Vitrebond. Taken together, we conclude that some components of glass ionomer cements show both genotoxic and cytotoxic effects. PMID- 16691347 TI - Phase evolution and sintering kinetics of hydroxyapatite synthesized by solution combustion technique. AB - Solution combustion technique has been used to prepare hydroxypatite (HAp) powder from calcium nitrate, di-ammonium hydrogen phosphate and citric acid precursors. Phase evolution has been studied as a function of calcination temperatures. The crystal structure, phase purity and stiochiometry of phase have been studied by Rietveld analysis of the calcined powder. It was observed that the prepared Hap powder was phase pure and stoichiometric. The sintering behaviour and sintering kinetics of the HAp compact has been studied by dilatometer. Activation energy for sintering has been calculated from the dilatometer results. Grain boundary diffusion was found to be the dominant densification mechanism during the initial stage of sintering. The activation energy for sintering (438 kJ/mol) was found to be in excellent agreement with reported value. PMID- 16691348 TI - Preparation of DHAQ-loaded mPEG-PLGA-mPEG nanoparticles and evaluation of drug release behaviors in vitro/in vivo. AB - This study describes the preparation and the evaluation of biodegradation monomethoxy (polyethylene glycol)-poly (lactide-co-glycolide)-monomethoxy (polyethyleneglycol) (mPEG-PLGA-mPEG, PELGE) nanoparticles (PELGE-NP) containing mitoxantrone (DHAQ) as a model drug. PELGE copolymers with various molar ratios of lactic to glycolic acid and different molecular weights and various content mPEG were synthesized by ring-opening polymerization. mPEG with weight-average molecular weight (Mw) 2,000 or 5,000 was introduced as a hydrophilic segment into a hydrophobic PLGA. A double emulsion method with dextran70 as stabilizer in the external aqueous phase was used to prepare the nanoparticles. The drug entrapment efficiencies were more than 80% and the mean diameters of the nanoparticles were less than 200 nm. Various PELGE was studied as biodegradable drug carriers and there in vitro/in vivo release profiles were examined. It was found that drug loading, polymer molecular weight, copolymer composition and end group modifications were critical factors affecting the in vitro/in vivo release properties. The amount of drug released increased as the mPEG contents increased and the molar ratios of lactic acid decreased in vitro. The intravenous (i.v.) administration of mPEG-PLGA-mPEG nanoparticles of DHAQ in mice resulted in prolonged DHAQ residence in systemic blood circulation compared to the intravenous administration of PLGA nanoparticles. PMID- 16691349 TI - Hydroxyapatite (HA) bone scaffolds with controlled macrochannel pores. AB - Hydroxyapatite (HA) macrochanneled porous scaffolds, with a controlled pore structure, were fabricated via a combination of the extrusion and lamination processes. The scaffold was architectured by aligning and laminating the extruded HA and carbon filaments. The macrochannel pores were formed by removing the carbon filaments after thermal treatments (binder removal and sintering). The porosity of the scaffolds was varied between 48 and 73% with a controlled pore size of approximately 450 microm, by adjusting the fractions of HA and carbon filaments. As the porosity was increased from 48 to 73%, the compressive strength decreased from 11.5 to 3.2 MPa. However, the osteoblast-like cell responses on the scaffold, such as the proliferation rate and alkaline phosphatase (ALP) activity, were significantly enhanced as the porosity was increased. PMID- 16691350 TI - Behaviour of MG-63 osteoblast-like cells on wood-based biomorphic SiC ceramics coated with bioactive glass. AB - The aim of this study was to test the in vitro cytotoxicity of wood-based biomorphic Silicon Carbide (SiC) ceramics coated with bioactive glass, using MG 63 human osteoblast-like cells, with a view to their application in bone implantology. To better understand the scope of this study, it should be taken into account that biomorphic SiC ceramics have only recently been developed and this innovative product has important properties such as interconnected porosity, high strength and toughness, and easy shaping. In the solvent extraction test, all the extracts had almost no effect on cellular activity even at 100% concentration, and cells incubated in the bioactive glass-coated SiC ceramics extracts showed a proliferation rate similar to that of the Thermanox control. There were no significant differences when the cellular attachment response of the cells on the wood-based biomorphic SiC ceramics, uncoated or coated with bioactive glass, was compared to the one exhibited by reference materials like Ti6Al4V and bulk bioactive glass. This fact looks very promising for biomedical applications. PMID- 16691351 TI - Mechanical and degradation behavior of polymer-calcium sulfate composites. AB - Calcium sulfate (CS) is one of the oldest bone graft materials still in use. Its main limitations are poor handling characteristics, poor mechanical properties, and a resorption rate that is too fast for some applications. The present study investigated the effect of viscous polymers, such as carboxymethylcellulose (CMC) and hyaluronan (HY), on the handling characteristics, mechanical properties, and degradation behavior of CS. CMC and HY were added to CS at concentrations from 1 10 wt%. Addition of CMC to CS at more than 4 wt% produced a putty-like material and decreased the density of the composite, while also increasing flexural and compressive strength at higher loadings. Incorporation of CMC produced a concentration-dependent increase in water absorption and degradation rate. At an equivalent loading, HY-containing CS composites showed better compressive strength than CS with CMC. Overall, addition of CMC or HY to CS resulted in composite materials with better handling characteristics and improved mechanical properties after set, however the degradation rate of the augmented materials was increased. These properties suggest that the enhanced CS materials may be useful in certain clinical situations, such as filling non-uniform bone defects and situations that require mechanical integrity of the bone graft substitute during implantation. PMID- 16691352 TI - Hydrothermal synthesis and nanostructure of carbonated calcium hydroxyapatite. AB - The influence of precursor concentration, pressure, temperature and time of hydrothermal synthesis on the development of calcium hydroxyapatite structure has been analyzed. The obtained results show that it is possible to adjust the conditions of hydrothermal synthesis from solutions of relatively high concentrations to obtain calcium hydroxyapatite nanopowders of well-defined structure. The relationship between the synthesis and the lattice parameters, as well as the crystallite size and the microstructure of synthesized hydroxyapatite has been established. The synthesized powders are preferentially carbonated hydroxyapatite of the B type in the form of agglomerates that accommodate two modal size pores of 1.5-10 and 50-200 nm. The structure of calcium hydroxyapatite particles consists of crystallites 8-22 nm in size, bound within prime particles, which size is between 10 and 63 nm, that in turn form bigger agglomerates 200 nm in size, which further cluster building up agglomerates 5-20 microm in size. PMID- 16691353 TI - Wound healing effect of silk fibroin/alginate-blended sponge in full thickness skin defect of rat. AB - Silk fibroin (SF) and alginate (AA) have been proved to be invaluable natural materials in the field of biomedical engineering. This study was designed to compare the wound healing effect of SF, AA and SF/AA-blended sponge (SF/AA) with clinically used Nu Gauze(TM) (CONT) in a rat full thickness wound model. Two circular skin wounds on the back of rat were covered with either of CONT, SF, AA or SF/AA. On the postoperative days of 3, 7, 10 and 14, residual wound area was calculated, and skin wound tissues were biopsied to measure the area of regenerated epithelium and collagen deposition as well as the number of proliferating cell nuclear antigen (PCNA)-immunoreactive cells. Half healing time (HT(50)) of SF/AA was dramatically reduced as compared with that of SF, AA or CONT. Furthermore, SF/AA significantly increased the size of re-epithelialization and the number of PCNA positive cells, whereas the effect of SF/AA on collagen deposition was not significantly different as compared with that of SF or AA. These results demonstrate that the wound healing effect of SF/AA is the best among other treatments including SF and AA, and this synergic effect is mediated by re-epithelialization via rapid proliferation of epithelial cell. PMID- 16691354 TI - Assessment of a three-dimensional measurement technique for the porosity evaluation of PMMA bone cement. AB - In vitro testing of bone cement has historically resulted in the belief that porosity should be minimised to help reduce the risk of prosthesis failure through aseptic loosening. Traditional porosity measurement techniques rely on the analysis of a two dimensional representation of a three dimensional structure. However, with an increasing interest in the number, size and distribution of pores in bone cement, the reliability of a two dimensional approach is questionable. The purpose of this study was to investigate the use of micro computed tomography (micro-CT) for the three dimensional measurement of bone cement porosity by comparison with two traditional techniques. Eighteen bone cement specimens were analysed for porosity using each technique. Levels of agreement between techniques were evaluated, and technique precision was assessed in terms of repeatability and sensitivity to changes in threshold. Micro-CT data was used to illustrate the effectiveness of predicting the porosity of a whole structure from a sample region; an approach often used with traditional techniques. In summary, poor agreement was found between all techniques. However, micro-CT was found to be significantly more repeatable and less sensitive to changes in threshold. The results demonstrated that porosity cannot be reliably determined using traditional techniques and that a large proportion of a specimen is required to provide an accurate porosity measurement. PMID- 16691355 TI - Optimization of preparation of DHAQ-loaded PEG-PLGA-PEG nonaparticles using central composite design. AB - Mitoxantrone (DHAQ)-loaded poly (ethylene glycol)-poly (lactic acid-co-glycolic acid) -poly (ethyleneglycol) (PELGE) nanoparticles (NP) were fabricated using an emulsification/solvent evaporation technique. A central composite design (CCD) was applied to evaluate the joint influence of three formulation variables: the amounts of polymer, concentration of the DHAQ, and the ratio of the organic phase (inner-phase) and the aqueous phase (outer-phase). In this study, we optimize the preparation technology on the basis of the single factor evaluation. The optimal conditions for the preparation of DHAQ-loaded nanoparticle were found to be: the concentration of PELGE was 9 mg/mL, the concentration of inner-phase of DHAQ was 27.5 mg/L, and the ratio of inner-phase/outer-phase was 8.5/1. The results showed that CCD is an ideal technique for formulation studies. The entrapment efficiency ratio (ER) was 90% and particle sizes are less than 500 nm. The nanoparticles, as examined by transmission electron microscopy (TEM), have a smooth and spherical surface. The DHAQ could be loaded into PELGE copolymers. In this study, the DHAQ nanoparticle-polymer delivery system was established by using PELGE polymers as carrier material. PMID- 16691356 TI - Laser induced multi-scale textured zirconia coating on Ti-6Al-4V. AB - A textured coating of zirconia on Ti-6Al-4V alloy was produced using pulsed laser based processing technique. Scanning electron microscope observations coupled with fractal analysis revealed the multi-scale nature of the textured coating. Both stylus based profilometric measurements and fractal analysis indicated non linear nature of the relationship between laser processing speed at constant pulse frequency (10 kHz) and roughness of the textured coating. The textured coatings produced with all the three processing speeds (40, 160, 290 cm/min) were fractal over certain length scales. Processing at 40 cm/min resulted in structures that are fractal across a large number of length scales where as higher processing speeds resulted in fractality over fewer length scales. The processing speed influenced the zirconia content in the coating and the phase transformation within Ti-matrix of the coating. Within the coating, while zirconia content decreased the amount of retained beta-Ti increased with increase in processing speed. Such physical and chemical transformations are desired in a titanium bio-implant for effective contact with protein, cells and tissues at various length scales and its effective chemical performance in bio-environment. PMID- 16691357 TI - Bioactive ceramic composites sintered from hydroxyapatite and silica at 1,200 degrees C: preparation, microstructures and in vitro bone-like layer growth. AB - Bioceramic composites were synthesized by sintering the powders of hydroxyapatite (HAp) mixed directly with additive of 0.5, 1.0, 2.0, 5.0 and 10 wt.%SiO(2), respectively, at 1,200( composite function)C. X-ray diffraction (XRD) analysis indicated that the phase transformation from HAp to tricalcium phosphate (TCP) comprising alpha-TCP and Si-TCP occurred and became more prominent with the addition of SiO(2) and the increase in SiO(2) content. The observations of their surface microstructures showed that the addition of SiO(2) suppressed the grain growth and promoted the formation of crystalline-glassy composites denoted HAp + TCP/Bioglass. As the SiO(2) content is as high as 5 wt.%, the composite made a feature of crystalline clusters with different sizes consisting of HAp and TCP grains surrounded by the matrix of glassy phase. Furthermore, the dependence of in vitro bioactivity of these composites on the SiO(2) content was biomimetically assessed by determining the changes in surface morphology, i.e., bone-like apatite layer growth, after soaking in an acellular stimulated body fluid (SBF) for 3 days at 36.5( composite function)C. It was found that the HAp-SiO(2) composites showed a much faster bone-like layer growth than pure HAp, and the propensity of composites to exhibit a better bioactivity was getting more notable with increasing SiO(2) content, except for the case of the highest content of 10 wt.%. It was believed that the formation of the bone-like layer on the surfaces of these bio-composites is closely related to the increasingly provided silanol groups and transformed TCP phase in materials associated with the content of SiO(2) added. PMID- 16691359 TI - Polypyrimidine-tract-binding protein is a component of the HCV RNA replication complex and necessary for RNA synthesis. AB - The machinery for hepatitis C virus (HCV) RNA replication is still poorly characterized. The relationship between HCV RNA replication and translation is also not clear. We have previously shown that a cellular protein polypyrimidine tract-binding protein (PTB) binds to HCV RNA at several different sites and modulates HCV translation in several ways. Here we show that PTB also participates in RNA replication. By bromouridine triphosphate (BrUTP) labeling and confocal microscopy of cells harboring an HCV replicon, we showed that the newly synthesized HCV RNA was localized to distinct structures in the cytoplasm, which also contain PTB. Membrane flotation analysis demonstrated that a fraction of cytoplasmic PTB was associated with a detergent-resistant membrane (DRM) structure consisting of lipid rafts, which also contained HCV nonstructural proteins and the human vesicle-associated membrane protein-associated protein (hVAP-33). PTB in the DRM was resistant to protease digestion, but became sensitive after treatment with the raft-disrupting agents. PTB in the DRM consisted of multiple isoforms and the brain-specific paralog. By using small interfering RNA (siRNA) of PTB, we showed that silencing of the endogenous PTB reduced the replication of HCV RNA replicon. In a cell-free, de novo HCV RNA synthesis system, HCV RNA synthesis was inhibited by anti-PTB antibody. These studies together indicated that PTB is a part of the HCV RNA replication complex and participates in viral RNA synthesis. Thus, PTB has dual functions in HCV life cycle, including translation and RNA replication. PMID- 16691360 TI - Virology- and immunology-based gene therapy for cancer. AB - Current strategies for cancer gene therapy consist mainly of direct inhibition of tumor cell growth and activation of systemic host defense mechanisms. Conventional chemotherapy and radiotherapy, even considered to be temporally suppressing tumor growth, suppress immune responses; therefore, we examined potential clinical feasibility of virus-mediated tumor destruction, which can rather enhance immunity. We showed that human tumors were more susceptible to adenoviruses (Ad) in which the E1A expression was controlled by a putative tumor promoter than normal cells, and that a replication of the Ad was greater in tumor cells than in normal cells. We also demonstrated that the intratumoral injection of the Ad bearing a tumor promoter inhibited the subsequent tumor growth in vivo. The E1A expression was detected in the tumors injected with the Ad but not in non tumorous tissues of the same mice. The Ad modified to show the regulated E1A expression is thereby oncolytic in nature. Antitumor immune responses are initiated after the acquisition of putative tumor antigen(s) by dendritic cells (DCs); therefore, enhanced antigen presentation is a crucial step for the early phase of cell-mediated immunity. Destruction of tumors can release the tumor antigens and DCs come to recognize them thereafter. We found that the stimulation of Fas expressed on DCs with Fas ligand (FasL) did not induce apoptosis of DCs but rather enhanced the antigen presentation. Activation of DCs induced production of a number of cytokines, and we showed that the interleukin-12 family secreted from tumors could induce systemic antitumor immunity. We presume that the administration of oncolytic Ad, which can destroy local tumors and subsequently make the putative tumor antigen(s) released from the tumors, stimulation of DCs with the Fas/FasL signal pathway and secretion of DCs-derived cytokines coordinately produce synergistic antitumor effects and that a combinatory application of these procedures can be a possible therapeutic strategy for cancer treatment. PMID- 16691361 TI - Beliefs about medicines among Swedish pharmacy employees. AB - OBJECTIVE: The aim of this study was to describe the beliefs about medicines among pharmacy employees. A further aim was to analyse whether these beliefs were associated with any background characteristics, such as age, professional category or medication use. METHOD: The study subjects were pharmacy employees at 24 community pharmacies in Goteborg, Sweden. The participating pharmacies had a total of 372 employees (pharmacists, dispensing pharmacists, and pharmacy technicians). Data was collected at the weekly pharmacy information meetings with a questionnaire comprising background questions and the Beliefs about Medicines Questionnaire (BMQ). The general part of the BMQ was used. For each statement in the BMQ, respondents marked their degree of agreement on a five-point Likert scale (1=strongly disagree, 2=disagree, 3=uncertain, 4=agree and 5=strongly agree). MAIN OUTCOME MEASURE: The three subscales of BMQ General: General Harm, General Overuse and General Benefit. RESULTS: The majority of the 292 respondents were dispensing pharmacists. More than half of the respondents were aged 45 years or older and had worked in a pharmacy for 20 years or more. Compared to the other professional categories, a higher proportion of dispensing pharmacists stated that they currently used traditional medicines. The pharmacy employees had a mean score for General Benefit of 4.31 and a mean score for General Harm of 1.81. Pharmacists and dispensing pharmacists in general regarded medicines as somewhat more beneficial, whereas pharmacy technicians viewed medicines as slightly more harmful. Those who had worked in a pharmacy for 30-34 years regarded medicines as less harmful compared to those who had worked 0-4 years. Compared to non-users, current users of prescription drugs regarded medicines as more beneficial. When controlling for background characteristics, no confounders were detected for any of the three subscales (ANCOVA analyses). CONCLUSION: Results of the study of 292 Swedish pharmacy employees show that they regard medicines as beneficial rather than harmful and that there are differences in beliefs between the professional categories. However, the reasons for these differences remain unclear. Nevertheless, the positive beliefs among pharmacy employees may have an impact on the communication with clients and, eventually, clients' adherence to medicines. PMID- 16691362 TI - Knowledge and use of complementary and alternative medicine among British undergraduate pharmacy students. AB - AIM: Attitudes and practice concerning complementary and alternative medicine (CAM) are currently an area of considerable importance. However, little is known about the overall importance of CAM in groups of health care professionals. Therefore, the primary objective was to analyse the knowledge about and use of CAM in an ethnically diverse student population at the London School of Pharmacy. METHODS: A questionnaire was administered to students (264 completed questionnaires, response rate: 59.1%) and in-depth interviews were carried out with a sample of seven students. RESULTS: Forty-three percent reported using at least one type of CAM during the last 12 months. The types of CAM used that were most frequently mentioned include aromatherapy, Traditional Chinese Medicine (TCM) and medicinal herbalism. TCM was reported by students of Chinese origin and by students from other ethnic backgrounds. However, Ayurvedic (Indian) medicine and medicine from Africa were only used in cultures from which the users originated. The study shows that a large number of herbal medical products were commonly reported by students of pharmacy. CONCLUSION: The knowledge about and use of CAM is widespread among this group of future health care professionals and the students also expressed a strong interest in the topic. In-depth studies on the knowledge and use of CAM among other health care professionals and among ethnic minorities are urgently needed and may help to better manage the treatment of minor disorders as well as chronic diseases. PMID- 16691365 TI - Factors affecting accuracy of ventricular volume and ejection fraction measured by gated tl-201 myocardial perfusion single photon emission computed tomography. AB - The electrocardiogram-gated single photon emission computed tomography (SPECT) measurement of left ventricular end-diastolic volume, end-systolic volume and ejection fraction may contain substantial errors. We evaluated whether patient related factors affect the accuracy of left ventricular volume and ejection fraction measured by gated Tl-201 SPECT. A total of 518 patients without perfusion defects on Tl-201 SPECT or coronary artery disease were studied. Left ventricular volume and ejection fraction were measured from echocardiography and adenosine stress/redistribution gated Tl-201 SPECT using commercially available software packages (QGS and 4D-MSPECT). We identified factors affecting the accuracy of gated SPECT via multiple linear regression analysis of the differences between echocardiography and gated SPECT. Gated SPECT analyzed with QGS underestimated end-diastolic and end-systolic volume, and overestimated ejection fraction, but 4D-MSPECT overestimated all those values (P<0.001). Independent variables associated with increasing the difference in end-diastolic volume between echocardiography and gated SPECT were decreasing left ventricular end-diastolic wall thickness, decreasing body surface area, female sex and increasing end-diastolic volume (P<0.001). Those for end-systolic volume were decreasing left ventricular end-systolic wall thickness, female sex, and decreasing end-systolic volume (P<0.001). Increasing end-systolic wall thickness, male sex and decreasing age were independent determinants associated with an increased difference in ejection fraction (P<0.001). Adenosine stress SPECT showed significantly higher end-diastolic and end-systolic volume values and a lower ejection fraction than did redistribution SPECT (P<0.001). Patient-related factors affect the accuracy of left ventricular volume and ejection fraction measured by gated Tl-201 SPECT. Modification of gated SPECT measurements by taking account of these factors would lead to reduce systemic errors. PMID- 16691366 TI - Influence of the acetolactate synthase inhibitor metsulfuron-methyl on the operation, regulation and organisation of photosynthesis in Solanum nigrum. AB - The influence of the acetolactate synthase inhibitor metsulfuron-methyl on the operation of the photosynthetic apparatus was examined on 4-weeks-old climate chamber-grown Solanum nigrum plant. To have an indication on the relative performance of the photosynthetic apparatus of ALS-treated plants, the level of carbon dioxide (CO(2)) fixation, the relative quantum efficiency of photosystem I (Phi(PSI)) or photosystem II (Phi(PSII)) electron transport and leaf chlorophyll content were assessed for both control and treated plants at 2, 4 and 7 days after application of the herbicide. Results indicated a progressive inhibition of the level of CO(2) fixation, the relative quantum efficiency of photosystem I (Phi(PSI)) and II (Phi(PSII)) electron transport and the leaf chlorophyll content already 2 days after application of the herbicide. The linear relationship between the photosystem I and II was unaltered by herbicidal treatment and was sustained under conditions where large changes in pigment composition of the leaves occurred. It appears that the stress-induced loss of leaf chlorophyll is not a catastrophic process but rather is the consequence of a well-organised breakdown of components. Under photorespiratory and non-photorespiratory conditions, the relationship between the index of electron transport flow through photosystem I and II and the rate of CO(2) fixation is altered so that electron transport becomes less efficient at driving CO(2) fixation. PMID- 16691367 TI - Limitations on microalgal growth at very low photon fluence rates: the role of energy slippage. AB - The lower limits of photosynthetically useable radiation at which growth and photosynthesis can occur establish the lower boundaries for the extent of photolithotrophy in the biosphere. Photolithotrophic growth denotes the capacity to grow with photons as the sole energy input. Slippage in terms of photosynthetic energy conversion implies a less than theoretical stoichiometry of energy-transduction process(es) such as the dissipation of intermediates of O(2) evolution and of ATP synthesis (H(+)/e(-) and H(+)/ATP ratios). Slippage is particularly important in limiting the growth of photolithotrophic organisms at very low photon fluence rates. We found that Dunaliella tertiolecta and Phaeodactylum tricornutum avoid such reductions in photon use efficiency by increasing the size and number of their photosynthetic units, respectively, and by altering Q(A) reduction kinetics on the reducing side of PS II. P. tricornutum is also less susceptible to slippage in terms of the breakdown of intermediates in its O(2) evolution pathway than D. tertiolecta. Minimizing H(+) leakage through the CF(0)-CF(1) ATP synthetase (and other H(+ )porters) is also discussed briefly. In combination, strategies employed by P. tricornutum effectively allow it to grow and photosynthesize at lower rates of energy input than D. tertiolecta, consistent with our observations. Differences in the responses of the photosynthetic apparatus of these two marine microalgae are mechanistic and probably representative of evolutionary divergences associated with strategies for dealing with environmental perturbations. PMID- 16691368 TI - A comparison of the three isoforms of the light-harvesting complex II using transient absorption and time-resolved fluorescence measurements. AB - In this article we report the characterization of the energy transfer process in the reconstituted isoforms of the plant light-harvesting complex II. Homotrimers of recombinant Lhcb1 and Lhcb2 and monomers of Lhcb3 were compared to native trimeric complexes. We used low-intensity femtosecond transient absorption (TA) and time-resolved fluorescence measurements at 77 K and at room temperature, respectively, to excite the complexes selectively in the chlorophyll b absorption band at 650 nm with 80 fs pulses and on the high-energy side of the chlorophyll a absorption band at 662 nm with 180 fs pulses. The subsequent kinetics was probed at 30-35 different wavelengths in the region from 635 to 700 nm. The rate constants for energy transfer were very similar, indicating that structurally the three isoforms are highly homologous and that probably none of them play a more significant role in light-harvesting and energy transfer. No signature has been found in the transient absorption measurements at 77 K for Lhcb3 which might suggest that this protein acts as a relative energy sink of the excitations in heterotrimers of Lhcb1/Lhcb2/Lhcb3. Minor differences in the amplitudes of some of the rate constants and in the absorption and fluorescence properties of some pigments were observed, which are ascribed to slight variations in the environment surrounding some of the chromophores depending on the isoform. The decay of the fluorescence was also similar for the three isoforms and multi exponential, characterized by two major components in the ns regime and a minor one in the ps regime. In agreement with previous transient absorption measurements on native LHC II complexes, Chl b --> Chl a energy transfer exhibited very fast channels but at the same time a slow component (ps). The Chls absorbing at around 660 nm exhibited both fast energy transfer which we ascribe to transfer from 'red' Chl b towards 'red' Chl a and slow transfer from 'blue' Chl a towards 'red' Chl a. The results are discussed in the context of the new available atomic models for LHC II. PMID- 16691369 TI - Evidence for a plasmalemma-based CO2 concentrating mechanism in Laminaria saccharina. AB - A kinetic analysis of the photosynthesis inhibition by buffers allowed quantification of some components of the carbon concentrating mechanism (CCM) of the brown macroalga Laminaria saccharina. The CCM was based on the presence of acid regions outside the plasma membrane that increased the CO(2) concentration available for photosynthesis by 10-20 times above that of the bulk medium at alkaline pH. Furthermore, the results suggested that the CCM is located mainly on the cell membrane and not in the chloroplast, as suggested for most macroalgae. The degree of dissipation of the acid regions by a buffer was related to the buffer anion concentration (B(-)), estimated from the titration of the buffer from bulk medium pH to a pH endpoint value close to the first pK (a) of the carbonic acid system. A kinetic model describing the relationship between inhibition of photosynthesis by a buffer and B(-) was developed suggesting that buffers act as competitive inhibitors with IC(50) (the concentration of the buffer anion which reduces the reaction velocity by half) of 5.0 mol m(-3). This model can be used to estimate the inhibitory effect of any buffer on the photosynthesis of L. saccharina. Nevertheless, some buffers tested showed a lower effect than that predicted from the hyperbolic model suggesting that their strength as inhibitors depended on: (1) the pK (a) in relation to the first pK (a) of the carbonic acid system and (2) its molecular weight (i.e. its mobility). PMID- 16691370 TI - Enhanced cell surface CD44 variant (v6, v9) expression by osteopontin in breast cancer epithelial cells facilitates tumor cell migration: novel post transcriptional, post-translational regulation. AB - Osteopontin (OPN) is a glycosylated, secreted phosphoprotein that functions both as a cell attachment and chemotactic factor. Elevated expression of OPN confers enhanced metastatic ability on transformed cells, suggesting that OPN may contribute to the malignant progression of tumors. Migration of mammary carcinoma cells is stimulated by OPN via interactions with integrins and CD44 cell surface receptors. We hypothesized that OPN modulates specific CD44 isoform expression to facilitate breast cancer cell migration. The 21NT tumorigenic human breast cancer cell line was examined for regulation of CD44 expression at both the mRNA and protein levels in response to an engineered increase in OPN expression under CMV promoter control. Significant up-regulation of CD44s isoform mRNA expression was observed, but no change in CD44v6, v8, v9 or v10 mRNA levels. While there were elevated levels of CD44s, v6 and v9 protein at the cell surface, at the level of total cellular protein only CD44s and v6 were markedly increased. This suggests that OPN can regulate CD44 expression at both transcriptional and post transcriptional (both amount and localization of protein) levels. To validate the functional consequence of OPN regulation of CD44 expression, we demonstrate that OPN-mediated cell migration was reduced by exposure to a anti-pan CD44 antibody, and to anti-CD44v6 and anti-CD44v9 function-blocking antibodies. Our data provide evidence that in 21NT cells OPN enhances CD44s mRNA expression, increases cell surface expression of CD44 variant forms without a change in mRNA levels, and stimulates cell migration. PMID- 16691371 TI - [Old age policy and health]. AB - Proceeding from a comprehensive definition of health which differentiates between personal and environmental aspects and which is explicitly not restricted to the absence of disease, age-related changes in functional capacity and characteristic diseases are discussed. To maintain or re-establish independency is said to be much more important than in the case of younger age groups because of the high prevalence of chronic diseases in old age. Discussing the development of costs in the health care system it is demonstrated that a considerable amount of the increasing costs for treatment in higher age groups is due to the higher portion of those who will die in the following years. As a consequence, an explosion of health care costs might be a too pessimistic scenario. The hypothesis of a compression of morbidity is discussed as a more optimistic perspective on cost development. Three additional parts deal with problems of German nursing care insurance, propositions for an integrative concept of preventive and rehabilitative care, and tasks and challenges of palliative care. The final part of this contribution inquires into priorities of future political action on the societal level, the level of public supplies, the personal level, and the family level. PMID- 16691372 TI - [Acute generalized exanthematous pustulosis with confirmed type IV allergy. Report of 3 cases]. AB - Acute generalized exanthematous pustulosis (AGEP) is characterized by an acute disseminated eruption of multiple, non-follicular pustules. In addition, it can be associated with fever, elevated erythrocyte sedimentation rate, and leukocytosis. The pathogenesis of AGEP is still unknown; however, medications appear a likely cause in most cases. We report on 3 patients with AGEP demonstrating characteristic clinical and histologic features after taking amoxicillin, ampicillin, or diltiazem. All patients had positive patch test reactions to the suspected drugs. PMID- 16691374 TI - The future of imaging techniques for cancer patients in The Netherlands : A Delphi study. AB - There is a need in The Netherlands to understand the future prospects of imaging techniques in cancer care to deal with current and expected capacity decisions. For this, a Delphi research was performed with a panel of 35 noted experts. The panel anticipates that the application of imaging techniques (e.g., CT, MRI, PET) will gradually change; especially, their use will also include earlier stages, i.e., cancer screening and treatment. The increasing need and investments for these techniques will be partly balanced by new developments (e.g., faster technology). There will be a substantial decrease in the use of ultrasound, CT (except for screening), and conventional radiographic diagnostics. Radical new techniques and/or developments are not expected. The panel foresees increasing partial task substitution and subspecialization. Further, a profound impact on imaging techniques is expected from developments outside the imaging field such as digitalization, changing patient demands, and commercialization. PMID- 16691378 TI - Colorectal hepatic metastases: quantitative measurements using single-shot echo planar diffusion-weighted MR imaging. AB - The purpose of this study was to obtain quantitative measurements of the apparent diffusion coefficient (ADC1), flow insensitive apparent diffusion coefficient (ADC2) and perfusion fraction (F) of colorectal hepatic metastases using DWI and to compare these measurements with those obtained in liver parenchyma. Forty patients with 66 hepatic metastases from colorectal carcinoma were prospectively evaluated using DWI with three b values. Quantitative maps of the ADC1 (using b = 0, 150, 500 s/mm2 images), ADC2 (using b = 150, 500 s/mm2 images) and fractional variation (F) between ADC1 and ADC2, which reflects perfusion fraction, were calculated. The ADC1, ADC2 and F derived from metastases and liver parenchyma were compared. The mean ADC1 values of liver parenchyma and metastases were significantly higher than the mean ADC2 values (P < 0.0001, paired t-test). Colorectal metastases were found to have higher mean ADC1 and ADC2 values compared with liver (P < 0.0001, Mann-Whitney test). However, the estimated F was found to be lower in metastases compared to liver (P = 0.03, Mann-Whitney test). Colorectal hepatic metastases were characterised by higher ADC1 and ADC2 values, but lower F values compared to liver. PMID- 16691379 TI - The relation between size at birth and risk of type 1 diabetes is not influenced by adjustment for the insulin gene (-23HphI) polymorphism or HLA-DQ genotype. AB - AIMS/HYPOTHESIS: Associations have been described between higher birthweight and increased risk of type 1 diabetes, and of insulin (INS) and human leucocyte antigen (HLA) genotypes that protect against diabetes with larger size at birth. We studied simultaneously the effects of size at birth, INS and HLA genotypes on the risk of type 1 diabetes to test whether the relation between size at birth and risk of type 1 diabetes would be strengthened after adjustment for INS and HLA genotypes. SUBJECTS AND METHODS: We designed a population-based case-control study in Norway with 471 cases of childhood-onset type 1 diabetes and 1,369 control subjects who were genotyped for the INS -23HphI polymorphism (surrogate for INS variable number of tandem repeats) and HLA-DQ alleles associated with type 1 diabetes. Data on birthweight and other perinatal factors were obtained from the Medical Birth Registry of Norway by record linkage. RESULTS: The data fitted a multiplicative model for the protective INS class III allele both within the INS locus and for the model with INS- and HLA-DQ-conferred risk of type 1 diabetes. We found no overall significant association between weight or head circumference at birth and the risk of type 1 diabetes, and adjustment for INS and HLA genotype did not influence this result. There was also no evidence for association of INS or HLA with size at birth among control subjects. CONCLUSIONS/INTERPRETATION: In contrast to suggestions from previous indirect studies, direct adjustment for INS and HLA genotypes did not lead to a stronger relation between birthweight and the risk of type 1 diabetes. PMID- 16691383 TI - Comparison of clinical and laboratory characteristics of cases with polycystic ovarian syndrome based on Rotterdam's criteria and women whose only clinical signs are oligo/anovulation or hirsutism. AB - This study was an attempt to determine whether the hormonal and clinical profiles of polycystic ovarian syndrome (PCOS) or non-PCOS cases whose only admission signs were oligo/anovulation or hirsutism. This retrospective study comprised a total number of 118, age-matched, young Turkish women with initial admission signs and symptoms of menstrual disorders (MD) like oligo/anovulation or hirsutism. Of these, 66 cases were diagnosed as PCOS, based on 2003 Rotterdam criteria [presence of two of first three criteria such as oligo- and/or anovulation, signs of clinical hyperandrogenism (HA-c) and/or biochemical signs of hyperandrogenism (HA-b) and polycystic ovaries on ultrasonography after exclusion of specific identifiable disorders]. Fifty-two women were diagnosed as cases of oligo/anovulation or hirsutism before the era of PCOS Rotterdam's consensus criteria. These two PCOS and non-PCOS cases were evaluated in terms of body mass index (BMI), waist-to-hip ratio, serum FSH, LH, estradiol (E2), dehydroepiandrosterone sulphate (DHEAS), androstendione (A) 17 hydroxyprogesterone (17-HP), fasting insulin, C-peptide levels, sex hormone binding globulin (SHBG) and finally, ultrasonographic ovarian morphology. PCOS cases with unilateral and bilateral polycystic ovarian morphology on ultrasound scan were analyzed based on Rotterdam criteria. No statistically significant difference was detected among two groups, in terms of BMI, waist-to-hip ratio, serum FSH, LH, E2, fasting insulin, C-peptide levels (P > 0.05). However, blood levels of DHEAS, A and 17-HP were higher, whilst SHBG levels were remarkably lower (P = 0.008) in PCOS cases. Among PCOS group, hormonal and clinical characteristics did not differ, irrespective or uni- or bilaterality of ovarian morphology on ultrasonographic scan. Percentages of cases with androgenic alopecia, oily skin/acnea and increased ovarian volume were higher in PCOS group; whereas Ferriman-Gallwey score >/= 8 were similar between two groups. Total but not free testosterone remained high in PCOS group (P < 0.01). In both PCOS and non-PCOS cases, a linear correlation was apparent between BMI and insulin levels (r (s )= 0.69 and 0.32, P < 0.05, respectively). Among PCOS group, MD + HA-b + HA c (n = 40) was present in 60.6% of subjects, MD + HA-b (n = 12) in 18.2%, and MD + HA-c (n = 14) in 21.2%. The three phenotypes did not differ in mean BMI, waist to-hip ratio and biochemical characteristics. To conclude, non-PCOS women with only sign or symptom of oligo/anovulation or hirsutism had a more favorable endocrine milieu. These cases should be followed in vigilance in an aim to confront the development of short- and long-term adverse effects of impending PCOS in the future. Furthermore, different phenotypes of PCOS cases were clinically or biochemically similar in characteristics. PMID- 16691384 TI - Over one thousand patients with early stage endometriosis treated with the Helica Thermal Coagulator (HELICA): safety aspects. AB - STUDY OBJECTIVE: To assess the safety of the Helica Thermal Coagulator in the laparoscopic treatment of early stage endometriosis. DESIGN: Retrospective, observational. SETTINGS: The Princess Royal University Hospital, The Sloane and Chelsfield Park Hospitals, Kent, UK. PATIENTS: One thousand and sixty patients with early stage endometriosis. RESULTS: All patients were treated laparoscopically with the Helica Thermal Coagulator; a new laparoscopic device that combines electrical energy with helium for the treatment of endometriosis. No major bladder, ureteric or bowel injuries occurred. The only complication was a perforated vagina from the cutting probe during dissection of the cul-de-sac in a patient with a vaginal endometriotic nodule. CONCLUSION: The Helica Thermal Coagulator is a safe device for the laparoscopic treatment of endometriosis. PMID- 16691385 TI - Repair of large supraspinatus rotator-cuff defects by infraspinatus and subscapularis tendon transfers in a cadaver model. AB - Transosseous repair of a supraspinatus tendon (SSP) defect (Patte size II) can be difficult if the tendon is retracted and the muscle atrophied. In this situation alternatives are margin convergence techniques, local tendon transfers or distant tendon transfers in massive tears. The object of this study was to compare two local tendon transfers in terms of the feasibility of the shift, the area covered by the shift and the force needed to accomplish the shift. Thirteen fresh-frozen cadaver shoulders were used. First a supraspinatus defect extending to the apex of the humeral head (Patte size II) was created. Transosseous repair was attempted with the infraspintus (ISP) and with the subscapularis (SCP) in all cases; repair was successful in all ISP cases, while use of the SCP resulted in a successful repair in only 8 of the 13 (61.5%). A significantly (P=0.012) larger defect area was covered by the ISP transfer than by the SCP shift: 89.7+/-8.5% versus 31.2+/-31.1% of the original defect, respectively. The tensile force needed to accomplish the shift was significantly (P=0.004) lower when the ISP was used (15+/-11 N) than with the SCP (37.1+/-15 N). In this cadaver model the ISP shift proved more favourable than the SCP shift for covering a Patte size II SSP defect. PMID- 16691386 TI - "Sandwich technique" in the surgical treatment of primary complex fractures of the femur and humerus. AB - In this prospective study, our aim was to determine the clinical and radiographic outcomes of the surgical treatment of primary complex problem femoral and humeral shaft fractures treated by a new technique called "sandwich technique." A total of 45 patients with comminuted, complex and/or osteopoenic fractures of the femur or humerus (30 femoral and 15 humeral fractures) were treated using this technique. The patients were followed up for a mean of 25 months. In 24 (85.7%) of 28 patients with femoral fractures and in 14 (93.3%) of 15 patients with humeral fractures, union was achieved within 3-6 months of the operation (mean: 4.5 months). The total union rate was 88.4%. The pseudoarthrosis rate was 12%. There was no infection or implant failure seen during the follow-up period. The cortical allograft struts can be used to provide collateral support to weakened osteopoenic/osteoporotic bone. This technique provides a union rate of about 88% in osteoporotic and/or complex primary humerus or femur fractures. PMID- 16691387 TI - Deformity correction and lengthening of lower legs with an external fixator. AB - Deformities combined with shortening in 34 lower limb segments of 28 patients were treated with an Ilizarov external fixator or a Taylor spatial frame at the same level as the osteotomy. We compared an acute correction group (A) with a gradual correction group (G) in patients undergoing deformity correction followed by lengthening. We retrospectively examined the amount of deformity correction, length gained, distraction index, maturation index, and external fixation index in both groups. The mean age of subjects was 12.9 years for A, 17.9 years for G. The mean deformity correction was 17.8 degrees for A, 25.1 degrees for G. Mean lengthening was 5.5 cm for A, 5.0 cm for G. Mean distraction index was 16.4 days/cm for A, 10.6 days/cm for G (P<0.05). Mean maturation index was 40.5 days/cm for A, 29.5 days/cm for G (P=0.081). Mean external fixation index was 58.6 days/cm for A, 42.5 days/cm for G (P<0.05). The distraction index and external fixation index differences between the two groups were statistically significant. Gradual correction may represent a better approach than acute correction with the use of external fixators to treat deformity combined with shortening. PMID- 16691388 TI - A long-term follow-up of 60 Lord total hip arthroplasties in rheumatic disease: a mean follow-up of 14 years. AB - The aim of the study was to analyse the survivorship of 60 total hip arthroplasties using the cementless Lord prosthesis in 51 patients with inflammatory joint disease. Patients were operated on between the years 1985 and 1988. The mean follow-up time was 13.8 (4.0-18.6) years. During the follow-up, one deep infection was encountered, and seven patients died of causes unrelated to the hip replacement. Revision surgery or death of the patient was used as an end point. The overall survival was 88.1% [95% confidence interval (CI) 76.6 94.1] for the stem, and 64.3% (95% CI 50.6-75.1) for the cup at 15 years. Causes for revision surgery were loosening of the cup in 17 hips, loosening of both components in five hips, and one deep infection. PMID- 16691393 TI - A biophysical basis for the inter-spike interaction of spike-timing-dependent plasticity. AB - Although spike-timing-dependent plasticity (STDP) is well characterized when pre- and postsynaptic spikes are paired with a given time lag, how this generalizes for more complex spike-trains is unclear. Recent experiments demonstrate that contributions to synaptic plasticity from different spike pairs within a spike train do not add linearly. In the visual cortex conditioning with spike triplets shows that the effect of the first spike pair dominates over the second. Using a previously proposed calcium-dependent plasticity model, we show that short-term synaptic dynamics and interaction between successive back-propagating action potentials (BPAP) may jointly account for the nonlinearities observed. Paired pulse depression and attenuation of BPAPs are incorporated into the model through the use-dependent depletion of pre- and postsynaptic resources, respectively. Simulations suggest that these processes may play critical roles in determining how STDP operates in the context of natural spike-trains. PMID- 16691396 TI - Rituximab as first line treatment for MALT lymphoma of extraocular muscles. PMID- 16691392 TI - Functions of neuronal P2Y receptors. AB - Within the last 15 years, at least eight different G protein-coupled nucleotide receptors, i.e., P2Y receptors, have been characterized by molecular means. While ionotropic P2X receptors are mainly involved in fast synaptic neurotransmission, P2Y receptors rather mediate slower neuromodulatory effects. This P2Y receptor dependent neuromodulation relies on changes in synaptic transmission via either pre- or postsynaptic sites of action. At both sites, the regulation of voltage gated or transmitter-gated ion channels via G protein-linked signaling cascades has been identified as the predominant underlying mechanisms. In addition, neuronal P2Y receptors have been found to be involved in neurotoxic and neurotrophic effects of extracellular adenosine 5-triphosphate. This review provides an overview of the most prominent actions mediated by neuronal P2Y receptors and describes the signaling cascades involved. PMID- 16691397 TI - A case-control study of myelodysplastic syndromes in Belgrade (Serbia Montenegro). AB - The objective of the study was to investigate factors related to the occurrence of myelodysplatic syndromes (MDS) in the population of Belgrade (Serbia Montenegro). The case-control study was conducted during the period 2000-2003. The study group consisted of 80 newly diagnosed MDS patients and 160 sex- and age matched hospital controls with nonmalignant and noninfectious diseases. The disease categories in the control group were circulatory (51 patients, 32%), gastrointestinal (53 patients, 33%), and ophthalmological (56 patients, 35%) disorders. Conditional univariate and multivariate logistic regression analyses were applied. Multivariate analysis showed the following factors to be significantly related to MDS: exposure to chemicals (OR = 10.8, 95%CI 3.2-36.2, p = 0.0001), viral upper respiratory tract infections (twice a year or more, OR = 5.8, 95%CI 2.5-13.6, p = 0.0001), exposure to insecticides, pesticides and herbicides (OR = 5.2, 95%CI 1.8-15.1, p = 0.003), coffee (OR = 5.1, 95%CI 1.9 13.7, p = 0.001), and alcohol consumption (OR = 2.2, 95%CI 1.1-4.6, p = 0.033). The findings support the hypotheses that exposure to chemical agents, pesticides, insecticides, and herbicides, certain lifestyle factors (alcohol and coffee consumption), and frequent viral infections may be involved in the etiology of MDS, but these results should be confirmed by further investigations. PMID- 16691398 TI - Treatment results of chemoradiation therapy for localized aggressive lymphomas: A retrospective 20-year study. AB - In this study we analyzed our cases of localized aggressive lymphoma treated in our institution during the last 20 years to compare the finding of this study with those of previous studies. Forty patients with Ann Arbor stage I-II aggressive lymphoma were treated with 3-6 cycles of a CHOP regimen (cyclophosphamide, doxorubicin, vincristine, and prednisolone) and radiation therapy (30 or 40 Gy with involved field). Between 1985 and 2003, 40 patients with stage I (N = 25) or stage II (N = 15) disease were treated. Chemotherapy mainly preceded radiotherapy, although the sequence of radiotherapy and chemotherapy was determined by individual physicians and patients' choice. Median and mean age was 50.5 and 48.6 years, respectively, at the time of diagnosis, with a male to female ratio of 19:21. Analyses were undertaken to determine (1) response to treatment according to age, international prognostic index (IPI), lactate dehydrogenase (LDH) value, serum interleukin 2 receptor (sIL-2R) value, cell type, stage, extent of maximum local disease, with or without mediastinal lymph nodes, number of sites, anatomic distribution, and irradiation dose, and (2) relapse patterns. Complete follow-up was obtained in all patients. The follow up period of surviving 33 patients ranged from 24.7 to 180 months with a median of 69 and a mean of 72.7 months. A complete remission (CR) was achieved in 37 patients (93%). A study of relapse patterns after a CR showed that four patients had a first relapse within a radiation field and the other one patient had an extranodal distant relapse. Significant prognostic factors were not identified by multivariate analysis. Combined chemotherapy and radiation therapy is safe, highly effective, and probably curative for most patients with stage I-II aggressive lymphoma. PMID- 16691399 TI - Iron and pregnancy--a delicate balance. AB - The review focuses on iron balance during pregnancy and postpartum in the Western affluent societies. Iron status and body iron can be monitored using serum ferritin, haemoglobin, serum soluble transferrin receptors (sTfR) and the sTfR/ferritin ratio. Requirements for absorbed iron increase during pregnancy from 0.8 mg/day in the first trimester to 7.5 mg/day in the third trimester. Average requirement during the entire gestation is approximately 4.4 mg/day. Intestinal iron absorption increases during pregnancy, but women with ample body iron reserves have lower absorption than those with depleted reserves, so increased absorption is, in part, due to progressive iron depletion. Apparently, women do not change dietary habits when they become pregnant. Non-pregnant Scandinavian women have a median dietary iron intake of approximately 9 mg/day, i.e. more than 90% of the women have an intake below the recommended approximately 18 mg/day. Non-pregnant women have a low iron status, 42% have serum ferritin levels 70 microg/l corresponding to body iron of >or=500 mg. The association between high haemoglobin during gestation and a low birth weight of the newborns is caused by inappropriate haemodilution. In placebo-controlled studies on healthy pregnant women, there is no relationship between the women's haemoglobin and birth weight of the newborns and no increased frequency of preeclampsia in women taking iron supplements. PMID- 16691400 TI - Beta2-microglobulin concentrations in cerebrospinal fluid correlate with neuroimaging findings in newborns with symptomatic congenital cytomegalovirus infection. AB - OVERVIEW: In newborns with symptomatic congenital cytomegalovirus (CMV) infection, neuroimaging is the best available predictor of neurodevelopmental outcome. Cerebrospinal fluid (CSF) findings in congenital CMV infection have seldom been described. Neonates with central nervous system infections present high CSF Beta(2)-microglobulin (beta(2)-m) levels. OBJECTIVES: The objectives of this study were: (1) to determine whether CSF beta(2)-m is increased in newborns with symptomatic congenital CMV infection, and (2) to examine its correlation with neuroimaging findings. MATERIALS AND METHODS: Fourteen newborns with symptomatic congenital CMV infection admitted to La Paz Hospital from 1990 through 2004 underwent determination of CSF beta(2)-m. Ninety-three newborns, constituting the comparison group, underwent CSF beta(2)-m determination as part of a sepsis or meningo/encephalitis work-up, and at discharge had sterile cultures and normal neurological status. Neuroimaging findings were scored according to a semiquantitative system: (0) no abnormalities; (1) single punctate periventricular (PV) calcification and/or hyperechogenic areas in the thalamus and basal ganglia; (2) multiple discrete PV calcifications and/or ventriculomegaly; and (3) extensive PV calcifications and/or brain atrophy. DISCUSSION AND CONCLUSION: CSF beta(2)-m was increased in newborns with CMV infection (median 6.21 mg/L) compared with controls (1.68 mg/L) (P<.001). beta(2) m showed a correlation with neuroimaging scores (r (s)=0.753, P=.002). beta(2)-m was higher in patients who scored 2-3 (12.83 mg/L) than in patients who scored 0 1 (5.52 mg/L) (P=.028). CSF beta(2)-m is increased in newborns with symptomatic congenital CMV infection and correlates with neuroimaging abnormalities. beta(2) m appears to be an indicator of the severity of brain involvement in congenital CMV infection. PMID- 16691401 TI - Acute meningitis due to Brucella spp. AB - Brucella spp. should be considered in the differential diagnosis of acute meningitis of children living in areas where brucellosis is endemic. PMID- 16691402 TI - Infantile seizures and other epileptic phenotypes in a Chinese family with a missense mutation of KCNQ2. AB - INTRODUCTION: Benign familial infantile seizures (BFIS) is a form of idiopathic epilepsy characterized by clusters of afebrile seizures occurring around the sixth month of life and a favorable outcome. Linkage analysis has revealed that three chromosomal segments, 19q12-q13.1, 16p12-q12, and 2q23-31, are linked to this disorder. SUBJECTS AND METHODS: We report here a large Chinese family in which all 17 affected members had had infantile seizures with onset at age 2-4 months, with two of these also manifesting seizures later in life accompanied with either choreoathetosis or myokymia. Linkage analysis in this family confirmed a previous report of genetic heterogeneity in BFIS - since linkage was excluded at the above-mentioned known BFIS loci - and suggested a possible linkage to the KCNQ2 gene, which is believed to be a voltage gated potassium channel gene responsible for benign familial neonatal seizures (BFNS). RESULTS AND DISCUSSION: Sequencing of the KCNQ2 gene revealed that all 17 affected family members carried a heterozygous Gly-to-Val (G271V) mutation in the conserved pore region that resulted from a guanine-to-thymine transition in exon 5 of KCNQ2. The same mutation with a comparable localization in the KCNQ3 (G310V) gene has been found in BFNS patients. The same conserved amino acid was also found to be mutated in the KCNQ1 gene in a family with Long QT Syndrome. PMID- 16691403 TI - Delayed internal ophthalmoplegia and amblyopia following chickenpox. AB - INTRODUCTION: Chickenpox can be responsible for various ophthalmologic complications [4]. DISCUSSION: An internal ophthalmoplegia could be observed in this context and is responsible for a fixed pupil [2]. This complication is extremely rare and seems to occur mainly in young children during the two weeks following cutaneous eruption. We report an atypical observation of internal ophthalmoplegia due to chickenpox that occurs with a particularly prolonged interval. CONCLUSION: We would like to emphasize the consequences of the paralysis of accommodation associated with this tonic pupil on visual acuity. PMID- 16691404 TI - A comparison study of the prevalence of overweight and obese Italian preschool children using different reference standards. AB - INTRODUCTION: The number of overweight children is constantly increasing in both developed and underdeveloped countries. Most epidemiological surveys consider school age children with little attention being given to preschool age children. This study was undertaken to define the prevalence of being overweight or obese in a population of children 3-6 years of age, living in North-East Italy. We also identified differences when comparing body mass index (BMI) data against three different national and international standards. SUBJECTS AND METHODS: The study involved 258 children (124 males and 134 females) with an average age of 4.8 years. Weight and height were obtained for each child and BMI was calculated according to the formula weight (kg)/height (m)(2). The prevalence of overweight and obese subjects was then determined using the growth curves of the Centers for Disease Control and Prevention (CDC), the growth curves of the International Obesity Task Force (IOTF) and the curves published in 2003 by Luciano et al. RESULTS: All three methods gave similar, but not identical, estimates of the prevalence of overweight in both boys and girls. However, when determining the prevalence of obesity, the use of the CDC curves led to a significantly higher prevalence of obesity in both males and females when compared to the growth charts of the IOTF and Luciano et al. BACKGROUND: The use of the CDC growth curves leads to an increase in the prevalence of obesity when evaluating children under 5 years of age. The lower cut-off values inherent in the CDC charts, in contrast to the ones proposed by the IOTF, allow for earlier identification of a larger number of subjects with weight problems and therefore provide the potential for earlier intervention. PMID- 16691405 TI - Unusual presentation of intradural endodermal cysts in young children under 2 years of age. Report of two cases. AB - INTRODUCTION: Spinal endodermal cysts are congenital maldevelopmental lesions usually presenting in early adulthood. Whereas the diagnosis is rare in newborns and it is usually led by cardiopulmonary distress or other associated malformations, no characteristic clinical findings associated with these entities have been described in older patients where spinal endodermal cysts usually behave as intraspinal space occupying lesions. DISCUSSION: The authors report on two children under 2 years of age affected by cervical endodermal cysts that presented with a history of acute motor deficit following a laryngitis. The hypothesis about the pathogenesis is that the upper respiratory tract infection might have stimulated the secretion of the cyst lining, inducing growth of the cyst and the sudden appearance of the motor deficit. Because of the apparent lack of relation between the benign laryngitis and the motor deficit, the occurrence of such an association might have been underestimated in some of the previously reported cases. PMID- 16691406 TI - Epidural cervical abscess in a neonate. PMID- 16691407 TI - Cerebral sinovenous thrombosis and idiopathic nephrotic syndrome in childhood: report of four new cases and review of the literature. AB - INTRODUCTION: Nephrotic children are prone to develop thromboembolic complications secondary to an acquired hypercoagulable state. Cerebral sinovenous thrombosis (CSVT) is increasingly recognised in this population, but clinical characteristics and outcome are not well documented. PATIENTS AND METHODS: The database of the Canadian Pediatric Ischemic Stroke Registry (Toronto Site) containing prospectively enrolled children from 1992-2004 with CSVT identified four children with NS. A pooled literature analysis retrieved 17 additional cases reports. RESULTS: CSVT presented in the majority of cases during the first flare or within 6 months after the onset of NS and was found to occur more often in SSNS/SDNS (n=13) than in SRNS (n=4). Clinical manifestations were non-specific and consisted primarily of seizures (n=8) and signs of raised intracranial pressure (n=16). Imaging studies revealed a predilection for superior sagittal sinus involvement (n=21) and rare parenchymal lesions (n=4). The most consistent biological risk factors were a severe hypoalbuminaemia (n=14) and, to a lesser extent, decreased antithrombin (AT) levels (n=9/16). Deficiency of other coagulation inhibitors (protein S, protein C) was not identified. Inherited thrombophilia was documented in a single case, suggesting that acquired, more than genetic, coagulation factors are involved. Anticoagulation was safe, and the outcome was good in most patients, and no recurrence of thrombotic event was reported. DISCUSSION: In conclusion, CSVT is now a well-described complication of NS with potential morbidity. A high index of suspicion is required, especially in young children with NS presenting neurological symptoms. Reliable biological predictors of CSVT are lacking. PMID- 16691408 TI - Infections and vaccinations preceding childhood Guillain-Barre syndrome: a prospective study. AB - INTRODUCTION: We performed a prospective, multicentre study in children with Guillain-Barre syndrome (GBS), diagnosed according to international criteria, to investigate the frequency and aetiology of antecedent diseases. All infections and vaccinations occurring within a 6-week period prior to the onset of GBS were documented. MATERIALS AND METHODS: Stool cultures, standardised serological investigations and PCR analyses for 24 different infective agents were performed. Serological findings were regarded as significant if specific immunoglobulin (Ig)M or IgA antibodies were detected, if the IgM enzyme immunoassay or immunfluorescence assay findings were confirmed by immunoblot, if complement fixation test titres rose fourfold or if geometric titres were more than threefold higher than in uninfected control persons. Ninety-five children with GBS were included in the study over a 40-month period. Preceding events were reported in 82%. RESULTS: Microbiological studies carried out on 84 patients resulted in a probable diagnosis in 46 (55%). Coxsackieviruses (15%), Chlamydia pneumoniae (8%), cytomegalovirus (7%) and Mycoplasma pneumoniae (7%) were the most frequently involved agents. Serological evidence of a Campylobacter jejuni infection was found in six patients (7%). Eight children had been vaccinated during the 6 weeks preceding the onset of GBS; in six of these children concomitant infectious diseases were reported, and in one child the time between vaccination and GBS was extremely short. CONCLUSION: We conclude that, in contrast to adults, Campylobacter spp. does not seem to play a major role in childhood GBS in German-speaking countries. The aetiology of antecedent diseases is distributed over a wide spectrum of paediatric infectious diseases. Most of the children who had been vaccinated showed concomitant infectious diseases, thus obscuring the causative role for GBS. PMID- 16691409 TI - Development of an optimal lidocaine infusion strategy for neonatal seizures. AB - INTRODUCTION: Lidocaine is an effective drug for the treatment of neonatal convulsions not responding to traditional anticonvulsant therapy. However, one of the side-effects is a risk of cardiac arrhythmias. The aim of this study was to develop an optimal dosing strategy with minimal risk of cardiac arrhythmias. MATERIALS AND METHODS: As a first step, we studied 20 neonates during routine treatment of neonatal seizures with lidocaine. All were given a loading dose of 2 mg/kg in 10 min, followed by the continuous infusion of 6 mg/kg per hour for 12 h, 4 mg/kg per hour for 12 h and finally 2 mg/kg per hour for 12 h. Effectiveness, cardiac toxicity and lidocaine plasma concentrations were then determined. RESULTS: No cardiac arrhythmias were observed, and lidocaine was effective in 76% of the treatments. In most of the treatments (13 out of 20) maximal lidocaine plasma concentrations were >9 mg/L. Plasma levels >9 mg/L have been related to cardiac toxicity when used as an anti-arrhythmic drug in adults. It was of interest that all preterm infants showed high lidocaine plasma levels. Secondly, we developed the optimal dosing regimen, which was defined as an infusion regimen at which maximal lidocaine plasma concentrations are <9 mg/L. Simulations with the developed pharmacokinetic model indicated a reduction in the infusion duration of lidocaine at 6 mg/kg per hour from 12 to 6 h. Thirdly, the new lidocaine dosing regimen was evaluated. Fifteen neonates (16 treatments) were studied. No cardiac arrhythmias were observed, and lidocaine was effective in 78% of the treatments. In most of the treatments (11 out of 16) maximal lidocaine plasma concentrations were <9 mg/L. Again preterm infants showed relatively high lidocaine plasma levels. CONCLUSION: A new lidocaine dosing schedule was developed. This new regimen should have a lower risk of cardiac arrhythmias and appears to be as effective in term infants. For preterm infants the optimal regimen needs to be determined. PMID- 16691410 TI - Falls from highchairs. PMID- 16691411 TI - Acute pancreatitis after growth hormone treatment: disease or treatment linked? PMID- 16691412 TI - Regulation of cartilaginous ECM gene transcription by chondrocytes and MSCs in 3D culture in response to dynamic loading. AB - This study explored the biologic response of chondrocytes and mesenchymal stem cells (MSCs) to a dynamic mechanical loading regime. We developed a time efficient methodology for monitoring regional changes in extracellular matrix gene transcription using reporter promoter constructs. Specifically, transfected cells were homogenously distributed throughout agarose hydrogel constructs, and spatial and temporal gene expression and the ability to form functional ECM were analyzed in response to dynamic mechanical stimuli. Theoretical analyses were used to predict the physical signals generated within the gel in response to these loading regimes. Using a custom compression bioreactor system, changes in aggrecan and type II collagen promoter activity in transfected chondrocyte-laden cylindrical constructs were evaluated in response to a range of loading frequencies and durations. In general, aggrecan promoter activity increased with increasing duration of loading, particularly in the outer annulus region. Interestingly, type II collagen promoter activity decreased in this annular region under identical loading conditions. In addition, we explored the role of mechanical compression in directing chondrogenic differentiation of MSCs by monitoring short-term aggrecan promoter activity. As an example of long-term utility, a specific loading protocol was applied to MSC-laden constructs for 5 days, and the resultant changes in glycosaminoglycan (GAG) production were evaluated over a 4-week period. This dynamic loading regime increased not only short-term aggrecan transcriptional activity but also GAG deposition in long-term culture. These results demonstrate the utility of a new reporter promoter system for optimizing loading protocols to improve the outcome of engineered chondrocyte and MSC-laden cartilaginous constructs. PMID- 16691413 TI - Rapid growth of cartilage rudiments may generate perichondrial structures by mechanical induction. AB - Experimental and theoretical research suggest that mechanical stimuli may play a role in morphogenesis. We investigated whether theoretically predicted patterns of stress and strain generated during the growth of a skeletal condensation are similar to in vivo expression patterns of chondrogenic and osteogenic genes. The analysis showed that predicted patterns of compressive hydrostatic stress (pressure) correspond to the expression patterns of chondrogenic genes, and predicted patterns of tensile strain correspond to the expression patterns of osteogenic genes. Furthermore, the results of iterative application of the analysis suggest that stresses and strains generated by the growing condensation could promote the formation and refinement of stiff tissue surrounding the condensation, a prediction that is in agreement with an observed increase in collagen bundling surrounding the cartilage condensation, as indicated by picro sirius red staining. These results are consistent with mechanical stimuli playing an inductive or maintenance role in the developing cartilage and associated perichondrium and bone collar. This theoretical analysis provides insight into the potential importance of mechanical stimuli during the growth of skeletogenic condensations. PMID- 16691414 TI - The PTHrP-Ihh feedback loop in the embryonic growth plate allows PTHrP to control hypertrophy and Ihh to regulate proliferation. AB - Growth plate and long bone development is governed by biochemical signaling pathways of which the PTHrP-Ihh system is the best known. Other factors, such as BMPs, FGFs and mechanical loading, may interact with this system. This study aims at elucidating the relative importance of PTHrP and Ihh for controlling proliferation, and hypertrophy in fetal growth plate cartilage. We assessed the question why reduced Ihh expression leads to more pronounced effects on the number of non-hypertrophic cells and total bone formation, compared to PTHrP down regulation. Using few basic equations, constituted from literature data, this paper shows how the PTHrP-Ihh feedback system can control different aspects of tissue differentiation at distinct locations. In particular, it is shown that (mechanical or biochemical) perturbations will affect proliferation via Ihh related parameters, whereas changes in PTHrP-related parameters selectively interact with hypertrophy. This is contra-intuitive, since PTHrP acts to keep cells proliferating. In this context, the critical PTHrP level for keeping cells proliferating has been reconsidered. In addition, an explanation is provided for the aforementioned difference in effect between reduced Ihh and PTHrP expression. PMID- 16691415 TI - Effects of fibrinolytic inhibitors on chondrogenesis of bone-marrow derived mesenchymal stem cells in fibrin gels. AB - The objective of this study was to examine the effect of two fibrinolytic inhibitors, aprotinin and aminohexanoic acid, on chondrogenesis of rabbit bone marrow mesenchymal stem cells (BM-MSCs). Rabbit BM-MSCs were obtained from the tibias and femurs of New Zealand White rabbits. Cell-fibrin constructs were made by mixing a cell-fibrinogen (10(7) cells/ml; 40 mg/ml fibrinogen) solution with a thrombin (5 IU/ml) solution and then divided into four groups: aprotinin control, aprotinin + transforming growth factor beta (TGF-beta), aminohexanoic acid control, and aminohexanoic acid + TGF-beta. Each of these groups was further treated with three different concentrations of inhibitors and the TGF-beta groups were treated with 10 ng/ml of TGF-beta1. The chondrogenic gene expressions, DNA content, and glycosaminoglycan content of samples were analyzed after 14 days of culture. The aprotinin groups exhibited significantly higher levels of aggrecan gene expression and glycosaminoglycan content than the aminohexanoic acid groups. However, inhibitor neither influenced gene expression of type II collagen nor proliferation (i.e., DNA content) of BM-MSCs. These findings suggest that fibrinolytic inhibitors used to control degradation of fibrin clot may influence TGF-beta-induced chondrogenesis of BM-MSCs. PMID- 16691416 TI - Retropharyngeal lymph node metastasis from olfactory neuroblastoma: A report of two cases. AB - Olfactory neuroblastoma is a rare, malignant neoplasm arising from the olfactory epithelium. It has an aggressive biological behavior that is characterized by local recurrence, atypical distant metastasis, and poor long-term prognosis. The incidence of cervical lymph node metastasis in olfactory neuroblastoma is variable, and treatment modalities are controversial. Moreover, few reports have been published concerning retropharyngeal lymph node metastasis from olfactory neuroblastoma. We present two cases of olfactory neuroblastoma with retropharyngeal lymph node metastasis. In addition, we provided a review of the current literature regarding olfactory neuroblastoma and retropharyngeal lymph node metastasis from olfactory neuroblastoma. PMID- 16691417 TI - Re-use of pectoralis major myocutaneous flap for the treatment of recurrent hypopharyngeal stenosis following total laryngectomy and postoperative radiotherapy: a case report. AB - Management of recurrent hypopharyngeal stenosis following total laryngectomy and postoperative radiotherapy is a challenging problem. Because of previous surgery, free flaps may not always be performed. We report a case of recurrent hypopharyngeal stenosis after a first attempt of reconstruction with a pectoralis major myocutaneous pedicled flap (PMMPF) failed. The problem was solved by reusing the previous PMMPF as a rotary door flap according to Eliachar and suturing it to the prevertrebral fascia according to Spriano. The follow-up is now of 9 months without recurrence. This new technique is described and discussed in comparison with other possible options. PMID- 16691419 TI - Regulation of gene expression and cell division by Polo-like kinases. AB - Much scientific research has focused on characterising regulatory pathways and mechanisms responsible for cell integrity, growth and division. This area of study is of direct relevance to human medicine as uncontrolled growth and division underlies many diseases, most strikingly cancer. In cancer cells, normal regulatory mechanisms for growth and division are often altered, or even fail to exist. This review summarises the mechanisms that control the genes and gene products regulating cytokinesis and cell separation in the fission yeast Schizosaccharomyces pombe, as well as highlighting conserved aspects in the budding yeast Saccharomyces cerevisiae and higher eukaryotes. Particular emphasis is put on the role of gene expression, the Polo-like kinases (Plks), and the signal transduction pathways that control these processes. PMID- 16691418 TI - On the origin and functions of RNA-mediated silencing: from protists to man. AB - Double-stranded RNA has been shown to induce gene silencing in diverse eukaryotes and by a variety of pathways. We have examined the taxonomic distribution and the phylogenetic relationship of key components of the RNA interference (RNAi) machinery in members of five eukaryotic supergroups. On the basis of the parsimony principle, our analyses suggest that a relatively complex RNAi machinery was already present in the last common ancestor of eukaryotes and consisted, at a minimum, of one Argonaute-like polypeptide, one Piwi-like protein, one Dicer, and one RNA-dependent RNA polymerase. As proposed before, the ancestral (but non-essential) role of these components may have been in defense responses against genomic parasites such as transposable elements and viruses. From a mechanistic perspective, the RNAi machinery in the eukaryotic ancestor may have been capable of both small-RNA-guided transcript degradation as well as transcriptional repression, most likely through histone modifications. Both roles appear to be widespread among living eukaryotes and this diversification of function could account for the evolutionary conservation of duplicated Argonaute Piwi proteins. In contrast, additional RNAi-mediated pathways such as RNA directed DNA methylation, programmed genome rearrangements, meiotic silencing by unpaired DNA, and miRNA-mediated gene regulation may have evolved independently in specific lineages. PMID- 16691420 TI - Medulloblastoma: histopathologic and molecular markers of anaplasia and biologic behavior. AB - Large cell/anaplastic (LC/A) medulloblastoma (MB) is a recently recognized variant of medulloblastoma known to be associated with an advanced stage and a poor prognosis. Although Eberhart et al. suggested histopathologic grading of medulloblastoma in 2002, no consensus has been reached in terms of determining the criteria of an LC/A variant, and its biological behavior continues to be the subject of debate. We retrospectively analyzed 74 cases (range 0.25-15 years) of MB clinicopathologically using the criteria established by Eberhart et al. The LC/A variant was identified in 16 cases (22% of MB cases), five of which showed a poor outcome. Most LC/A variant cases revealed synaptophysin immunoexpression (75%), but no epidermal growth factor receptor (EGFR) expression. Expression of synaptophysin, NeuN, GFAP, p53, c-erbB2, and EGFR did not differ in LC/A and non LC/A variants. Seven of the 74 cases of medulloblastoma showed erbB2 amplification by FISH, four of which were LC/A variants. N-myc amplification was observed in only one LC/A variant, but no c-myc amplification was found. In patients younger than 10 years, the LC/A variant showed a significantly poorer outcome than the non-LC/A variant (P = 0.02), while no difference was found in older patients. Multivariate analysis revealed only metastasis on MRI and p53 expression, but not anaplasia as unfavorable prognostic factors. Our study suggests that prognostic implications of anaplasia in medulloblastoma are uncertain, and that the reproducibility of the histopathologic criteria of the LC/A variant should be reassessed before they can be applied in practical use. PMID- 16691423 TI - [Fracture of the proximal humerus in children and adolescents. The most overtreated fracture]. AB - BACKGROUND: There is great uncertainty among trauma surgeons regarding the correct treatment of rare proximal humerus fractures in children and adolescents, in spite of the great potential of the proximal humeral epiphysis for self correction, even in cases of gross malalignment. METHODS: Over a span of 6 years, we treated 52 children and adolescents with closed proximal humerus fractures, 45 of whom had no other injuries. In only three cases did we see an indication for operative treatment at our institution. Operations on another three were performed elsewhere. Forty two patients were available for follow-up exams. RESULTS: They all had good or very good results with regard to fracture healing (constant score) independently of age, sex, fracture morphology, and treatment. CONCLUSIONS: We believe it is particularly important to discuss therapy concepts with parents and the involved outpatient physicians in order to avoid unnecessary and stressful operations. PMID- 16691424 TI - Severe bilateral central serous chorioretinopathy in a black patient: 16 years follow-up. AB - PURPOSE: To describe the exceptionally severe, bilateral, sight-compromising course of central serous chorioretinopathy (CSC) in a black patient. DESIGN: Observational case report. METHODS: We reviewed the clinical and angiographic findings of a 50-year-old black male patient with severe bilateral chronic CSC. RESULTS: The first attack was recorded 16 years earlier and it was asymmetrical. In OD, only retinal pigment epithelium (RPE) alterations were detected, while in OS there was a large serous retinal detachment with two smaller RPE detachments. Visual acuity (VA) was 1.0 OD and 0.6 OS. Gradually, after multiple remissions and exacerbations, a huge area of atrophy occupied the posterior pole OS, leading to a dramatic decrease of VA (0.02). The lesions also progressed and remained active in OD (VA 0.2). CONCLUSIONS: CSC can be exceptionally severe, non-benign, sight-compromising, with multiple remissions and exacerbations during the lifetime. Indocyanine-green angiography is useful for the long-term follow-up in severe cases, showing lesions that are not obvious in fluorescein angiography or funduscopy. PMID- 16691425 TI - Nonhypotensive dose of beta-adrenergic blocker ameliorates capillary deficits in the hearts of rats with moderate renal failure. AB - Renal failure causes sympathetic overactivity and inadequate capillary growth in response to cardiomyocyte hypertrophy in experimental renal failure, as well as in uremic patients. In nonuremic animals, sympathetic overactivity was shown to suppress capillary growth. The purpose of this study was to examine whether blockade with alpha- and beta-adrenoblockers ameliorates the capillary deficit that was documented in the hearts of rats with moderate renal failure. Male Sprague-Dawley rats, 3 days after surgical ablation [subtotal nephrectomy (SNX)] or sham operation (sham), were treated with phenoxybenzamine, metoprolol, or a combination of both: After 12 weeks, the hearts were investigated using morphometric and stereologic techniques. The length density of myocardial capillaries was lower (p<0.05) in untreated SNX than in sham (2,786+/-372 vs 3,397+/-602 mm/mm3); the decrease was abrogated by metoprolol (3,305+/-624 mm/mm3), but not by phenoxybenzamin (2,628+/-480 mm/mm3). The intercapillary distance increased (p<0.05) in SNX (20.5+/-1.5 microm) and tended to be lower after metoprolol treatment (19.0+/-1.9 microm). The media area of intramyocardial arterioles was significantly higher in untreated SNX (1,158+/-1,343 vs 686+/-771 microm2 in sham). Metoprolol in nonhypotensive doses prevents the capillary deficit in the hearts of rats with moderate renal failure and presents an argument for an important role of sympathetic overactivity in the genesis of the capillary deficit in moderate chronic renal insufficiency. PMID- 16691428 TI - Computer-assisted learning in medicine. How to create a novel software for immunology. AB - Teaching medical issues is increasingly demanding due to the permanent progress in medical sciences. Simultaneously, software applications are rapidly advancing with regard to their availability and easy use. Here a novel teaching program is presented for immunology, which is one of the fastest expanding topics in medical sciences. The requirements of media didactics were transferred to this e-learning tool for German students. After implementation, medical students evaluated the software and the different learning approaches showed acceptance. Altogether this novel software compares favourably to other English e-learning tools available in the Internet. PMID- 16691427 TI - Cell death effects of resin-based dental material compounds and mercurials in human gingival fibroblasts. AB - In order to test the hypothesis that released dental restorative materials can reach toxic levels in human oral tissues, the cytotoxicities of the resin-based dental (co)monomers hydroxyethylmethacrylate (HEMA), triethyleneglycoldimethacrylate (TEGDMA), urethanedimethacrylate (UDMA), and bisglycidylmethacrylate (BisGMA) compared with methyl mercury chloride (MeHgCl) and the amalgam component mercuric chloride (HgCl2) were investigated on human gingival fibroblasts (HGF) using two different test systems: (1) the modified XTT test and (2) the modified H 33342 staining assay. The HGF were exposed to various concentrations of the test-substances in all test systems for 24 h. All tested (co)monomers and mercury compounds significantly (P<0.05) decreased the formazan formation in the XTT-test. EC50 values in the XTT assay were obtained as half maximum-effect concentrations from fitted curves. Following EC50 values were found (mean [mmol/l]; s.e.m. in parentheses; n=12; * significantly different to HEMA): HEMA 11.530 (0.600); TEGDMA* 3.460 (0.200); UDMA* 0.106 (0.005); BisGMA* 0.087 (0.001); HgCl2* 0.013 (0.001); MeHgCl* 0.005 (0.001). Following relative toxicities were found: HEMA 1; TEGDMA 3; UDMA 109; BisGMA 133; HgCl2 887; MeHgCl 2306. A significant (P<0.05) increase of the toxicity of (co)monomers and mercurials was found in the XTT-test in the following order: HEMA < TEGDMA < UDMA < BisGMA < HgCl2 < MeHgCl. TEGDMA and MeHgCl induced mainly apoptotic cell death. HEMA, UDMA, BisGMA, and HgCl2 induced mainly necrotic cell death. The results of this study indicate that resin composite components have a lower toxicity than mercury from amalgam in HGF. HEMA, BisGMA, UDMA, and HgCl2 induced mainly necrosis, but it is rather unlikely that eluted substances (solely) can reach concentrations, which might induce necrotic cell death in the human physiological situation, indicating that other (additional) factors may be involved in the induction of tissue (pulp) inflammation effects after dental restauration. PMID- 16691429 TI - The long-term efficacy and safety of new biological therapies for psoriasis. AB - Long-term therapy is often required for psoriasis. This article reviews the most recent long-term clinical data for biological agents that have been approved or for which late-stage development data have been released for the treatment of patients with moderate to severe plaque psoriasis. Efficacy data are available for up to five 12-week courses of alefacept (approximately 60 weeks of therapy), 36 months (144 weeks) of continuous efalizumab, 48 weeks of continuous etanercept, and 50 weeks of bimonthly infliximab. Data sources include publications, product labeling, and posters presented at recent international scientific meetings. Alefacept appears to continue to be efficacious over multiple treatment courses for some responsive patients. The efficacy of efalizumab achieved during the first 12-24 weeks of therapy appears to be maintained or improved through at least 60 weeks of continuous treatment. The efficacy of etanercept appears to be maintained through at least 48 weeks of continuous treatment. Infliximab demonstrates a high response rate soon after initiation, which appears to be maintained through 24 weeks but declines modestly with therapy out to 50 weeks. After 48 weeks, approximately 60% of efalizumab treated and 45% of etanercept-treated patients remaining on therapy achieved > or =75% improvement from baseline in Psoriasis Area and Severity Index, as did 70.5% of infliximab patients who did not miss more than two infusions. Safety data suggest that these agents may be used for long-term administration. Long-term data from psoriasis trials continue to accumulate. Recent data suggest that biological therapies have efficacy and safety profiles suitable for the long-term treatment of patients with moderate to severe psoriasis. PMID- 16691430 TI - Lack of association with TNF-alpha-308 promoter polymorphism in patients with vitiligo. AB - Vitiligo is an acquired depigmentary disorder of the skin, characterized by incomplete penetrance, multiple susceptibility loci and genetic heterogeneity. An immunologic hypothesis is currently advanced as a possible pathogenesis of vitiligo. The cytokines have an important role in pathogenesis of autoimmunity in which tumor necrosis factor-alpha (TNF-alpha), a paracrine inhibitor of melanocytes, is especially important. Several single-nucleotide polymorphisms (SNP) have been identified in the human TNF gene promoter. The polymorphism at position -308 (TNF-308), which involves substituting G for A and designing the AA genotype, leads to a higher rate of TNF gene transcription than the wild-type GG genotype in in vitro expression studies. It has also been linked to increased susceptibility to several chronic metabolic, degenerative, inflammatory and autoimmune diseases. Therefore, we investigated the TNF-alpha-308 SNP in patients with vitiligo. We examined 61 patients with vitiligo. Healthy age-, ethnically- and sex-matched individuals (n = 123) served as controls. Polymerase chain reaction amplification was used for analysis of the polymorphism at position -308 in promoter of TNF-alpha gene. We found that the distribution of TNF-alpha genotypes in vitiligo patients did not differ from that in control subjects (P > 0.05). Moreover, there was no association between TNF-alpha genotypes and types of vitiligo. In conclusion, we suggest that TNF-alpha-308 SNP is not a genetic risk factor for vitiligo susceptibility. PMID- 16691433 TI - Genetic diversity of Chinese water deer (Hydropotes inermis inermis): implications for conservation. AB - The Chinese water deer (Hydropotes inermis inermis) is endemic to China. Historically, the species was widely distributed, but now, habitat loss and poaching have reduced its range and number drastically. In order to provide useful information for its conservation, we have investigated the genetic diversity and population structure of the Chinese water deer by analyzing the 403 bp fragment of the mitochondrial DNA (mtDNA) control region (D-loop). Eighteen different haplotypes were detected in 40 samples. Overall, Chinese water deer have a relatively high-genetic diversity compared to other rare cervid species, with a haplotype diversity of 0.923+/-0.025 and nucleotide diversity of 1.318 +/- 0.146%. No obvious phylogenetic structure among haplotypes was found for samples of different origin. An analysis of molecular variance (AMOVA) showed significant differentiation between the Zhoushan and the mainland population (F(ST)= 0.088, P < 0.001; Phi( ST ) = 0.075, P = 0.043), which suggests that exchanges of individuals between Zhoushan and the mainland should be avoided. We also recommend that a breeding center be set up for the mainland population. PMID- 16691434 TI - Isolation and sequence analysis of the Sox-1, -2, -3 homologs in Trionyx sinensis and Alligator sinensis having temperature-dependent sex determination. AB - Members of the Sox gene family are characterized by an HMG-box that shows sequence similarity with that of the mouse testis-determining gene Sry. Using degenerate primers PCR, seven and eight HMG-box motifs of Sry-related genes were cloned and sequenced from genomic DNA of Trionyx sinensis (termed TS41-47) and Alligator sinensis (AS41-48) with TSD (temperature-dependent sex determination). Among 15 Sry-related genes, TS41, TS42, AS41, and AS42 shared 80, 72, 81, and 79% amino acid identity, respectively, with each HMG-box domain of the mouse Sox-1, 2, and -3 genes by Blast analysis. Molecular phylogenetic analysis showed that the clustering of TS41-42 and AS41-42 was distant to the clustering of the nonreptilian vertebrate Sox-1, -2, -3 homologs, including fish, amphibian, bird, and mammals. The amino acid identity among TS41-42, AS41-42, and the nonreptilian vertebrate Sox-1, -2, -3 homologs is lower than identities among the Sox-1, -2, 3 homologs, suggesting that the sequence changes in TS41-42 of Trionyx sinensis and AS41-42 of Alligator sinensis might have occurred after the diversification of amniotes. PMID- 16691435 TI - Identification and characterization of LIW, a novel domain involved in animal NCKIPSDs and some uncharacterized fungal proteins. PMID- 16691436 TI - Genetic variations of 13 indigenous Chinese goat breeds based on cytochrome B gene sequences. AB - Phylogenetic relationships among and genetic variability within 13 Chinese indigenous goat breeds and Boer goat were analyzed using cytochrome b gene sequences. There were 44 variable sites found in a 642 bp sequence, and 46 Cyt b haplotypes were subsequently defined. The phylogeny analysis of haplotypes in combination with goat Cyt b sequences from GenBank shows that Chinese goats are obviously separated from wild goats and might come from Capra aegagrus. Further analysis indicated that indigenous Chinese goats might descend from at least two lineages; most of the individuals analyzed could be classified into lineage A as defined by Luikart, but five other goats were of uncertain lineage. The Tibet plateau is a possible place of origin for Chinese goats. The neighbor-joining tree based on pairwise differences among populations shows that most Tibetan goats, except the Middle Tibet type, cluster closely with North China goats, and then with South China goats. This result confirms that differences in genetic structure exist among goats in different geographic locations. Nucleotide diversity varied among populations. Tibet and North China goats had higher genetic diversity than South China goats. The fixation index (F (st)=87.72%) suggested that most of the total genetic variation was due to variation within populations. In addition, the results indicate that Cyt b gene sequence information alone might not be enough for phylogeny analysis among breeds within species, as shown by fewer polymorphic sites and lower bootstrap values on the neighbor-joining tree. PMID- 16691437 TI - Phrasing effects in comprehending PP constructions. AB - The role of prosodic phrasing in sentence comprehension was investigated by means of three different tasks, namely auditory word monitoring (Experiment 1), self paced reading (Experiment 2) and cross-modal comparison (Experiment 3). In all three experiments a critical prosodic unit or frame comprising a determiner, a noun and a Prepositional Phrase (PP) was preceded or surrounded by two context prosodic units (frames) whose length was varied. The participants' tendency to interpret the critical sequence as forming a single syntactic constituent (noun complement interpretation of the PP) as opposed to two distinct syntactic constituents (verb-complement interpretation of the PP) was found to depend on the relative length of the critical and context prosodic units (frames). As a whole these results are consistent with the notion that phrasing effects occur in a retroactive way, after part of the utterance has been processed. PMID- 16691438 TI - Long-term changes in spinal cord evoked potentials after compression spinal cord injury in the rat. AB - 1. After traumatic spinal cord injury (SCI), histological and neurological consequences are developing for several days and even weeks. However, little is known about the dynamics of changes in spinal axonal conductivity. The aim of this study was to record and compare repeated spinal cord evoked potentials (SCEP) after SCI in the rat during a 4 weeks' interval. These recordings were used: (i) for studying the dynamics of functional changes in spinal axons after SCI, and (ii) to define the value of SCEP as an independent outcome parameter in SCI studies. 2. We have used two pairs of chronically implanted epidural electrodes for stimulation/recording. The electrodes were placed below and above the site of injury, respectively. Animals with implanted electrodes underwent spinal cord compression injury induced by epidural balloon inflation at Th8-Th9 level. There were five experimental groups of animals, including one control group (sham-operated, no injury), and four injury groups (different degrees of SCI). 3. After SCI, SCEP waveform was either significantly reduced or completely lost. Partial recovery of SCEPs was observed in all groups. The onset and extent of recovery clearly correlated with the severity of injury. There was good correlation between quantitated SCEP variables and the volumes of the compressing balloon. However, sensitivity of electropohysiological parameters was inferior compared to neurological and morphometric outcomes. 4. Our study shows for the first time, that the dynamics of axonal recovery depends on the degree of injury. After mild injury, recovery of signal is rapid. However, after severe injury, axonal conductivity can re-appear after as long as 2 weeks postinjury. In conclusion, SCEPs can be used as an independent parameter of outcome after SCI, but in general, the sensitivity of electrophysiological data were worse than standard morphological and neurological evaluations. PMID- 16691439 TI - Stress triggered changes in gene expression in adrenal medulla: transcriptional responses to acute and chronic stress. AB - 1. Stress elicits activation of several transcription factors involved in the regulation of catecholamine biosynthetic enzyme gene expression depending on its duration or repetition. However, the dynamic of the conversion of transient transcriptional activation with acute stress to sustained changes in transcription in response to repeated exposure to stress in adrenomedullary catecholaminergic systems is not clear. 2. Here, we analyzed changes in levels of phospho-CREB (P-CREB), phospho-ERK1/2 (P-ERK1/2) and Fra-2 by Western Blot analysis in adrenal medulla of Sprague Dawley male rats exposed to single or repeated immobilization stress (IMO). For single stress, rats were immobilized for 5 min, 30 min, or 2 h and sacrificed immediately afterwards. In the repeated stress conditions, animals were immobilized for 2 h daily on each consecutive day prior to the final day (day 2 for 2x IMO, day 6 for 6x IMO) in which the rats were immobilized for a session lasting 5 min, 30 min or 2 h. There were two control groups, an absolute control (AC) not exposed to stress, and a handled control (HC) gently handled daily for 6 days. 3. Phosphorylation of CREB was rapid and elevated at the earliest time examined, even with single stress. However, with a second daily episode of stress the increase in P-CREB was observed for at least the entire duration of the stress. In contrast, phosphorylation of ERK1/2 was only significant after brief exposure to a single IMO. The elevation of Fra-2 protein level was slower, but was significant after 2 h of a single IMO. With repeated IMO, there were marked elevations of Fra-2 throughout the 2 h IMO, which were especially pronounced at the end of the immobilization. 4. The transient nature of the phosphorylation of CREB may be responsible for the short-lived induction of transcription of catecholamine biosynthetic enzymes after brief exposure to a single immobilization stress. The sustained phosphorylation of CREB throughout the repeated stress coupled with induction of Fra-2 may mediate the longer lasting responses to repeated stress. PMID- 16691440 TI - Cell proliferation in the adult rat rostral migratory stream following exposure to gamma irradiation. AB - One of the few areas of the adult CNS, that are known to be competent for neuronal proliferation, is the subventricular zone (SVZ) lining the brain lateral ventricles. Cells proliferating in the SVZ migrate along a defined pathway, the rostral migratory stream (RMS), where their proliferation continues until reaching the olfactory bulb. 1. In relation to the fact that brain is, in general, regarded as a radioresistant organ composed from non dividing cells, the aim of the present study was to investigate effect of ionizing radiation on proliferating cell numbers in the RMS of adult rats. 2. Male Wistar rats were investigated 25 and 80 days after whole body gamma irradiation with the dose of 3 Gy. Dividing cells were labeled by bromodeoxyuridine (BrdU). BrdU-positive cells were counted by Disector program. The mean number of BrdU(+) cells in the whole RMS and in its individual parts (vertical arm, elbow, and horizontal arm) was evaluated. 3. Temporary increase in proliferating cell number (by 30%) was seen in the whole RMS at the 25th day after irradiation. 4. The most expressive increase occurred in the vertical arm (by 60%) and elbow (about 37%). The values reduced till the 80th day after exposure. Our results show that ionizing irradiation significantly influences the extent of cell proliferation and migration in the adult rat RMS. PMID- 16691441 TI - Gene expression of phenylethanolamine N-methyltransferase in corticotropin releasing hormone knockout mice during stress exposure. AB - AIMS: Epinephrine (EPI) synthesizing enzyme phenylethanolamine N methyltransferase (PNMT, EC 2.1.1.28) is primarily localized in the adrenal medulla (AM). We have recently described existence of the PNMT gene expression in cardiac atria and ventricles and in sympathetic ganglia of adult rats and mice. The aim of the present work was to study regulation of the PNMT gene expression in corticotropin-releasing hormone knockout mice (CRH KO) and matched control wild-type mice (WT) under normal and stress conditions. METHODS: Levels of the PNMT mRNA were determined by RT-PCR; PNMT immunoprotein and protein of transcription factor EGR-1 by Western Blot. Plasma EPI and corticosterone (CORT) levels were determined by radioenzymatic and RIA methods. Immobilization (IMMO) was used as a stressor. RESULTS: Stress-induced increases in the PNMT mRNA and protein levels observed in WT mice were almost completely absent in CRH KO mouse adrenal medulla, stellate ganglia, and cardiac atria, while ventricular PNMT mRNA elevation was not CRH-dependent. Plasma EPI and CORT levels were markedly reduced in CRH KO compared to WT mice both before and after the stress. Levels of EGR-1, crucial transcription factor for regulation of the PNMT were highly increased in stressed WT and CRH KO mice in cardiac areas, but not in the adrenal medulla. CONCLUSIONS: Data show that the CRH deficiency can markedly prevent immobilization-triggered induction of the PNMT mRNA and protein levels in the adrenal medulla and stellate ganglia. Reduced plasma epinephrine and corticosterone levels and adrenal medullary EGR-1 protein levels in CRH knockout versus WT mice during stress indicate that the HPA axis plays a crucial role in regulation of the PNMT gene expression in these organs. Cardiac atrial PNMT gene expression with stress is also dependent on intact HPA axis. However, in cardiac ventricles, especially after the single stress exposure, its expression is not impaired by CRH deficiency. Since cardiac EGR-1 protein levels in CRH KO mice are also not affected by the single stress exposure, we propose existence of different regulation of the PNMT gene expression, especially in the cardiac ventricles.Overall, our findings reveal that the PNMT gene expression is regulated through the HPA in both sympathoadrenal system and the heart and also via EGR-1 in the adrenal medulla, but apparently not in the heart. Regulation of the PNMT gene expression in various compartments of heart includes both corticosterone-dependent and independent mechanisms. PMID- 16691442 TI - Spatiotemporal organization of Ras signaling: rasosomes and the galectin switch. AB - 1. Ras signaling and oncogenesis depend on the dynamic interplay of Ras with distinctive plasma membrane (PM) microdomains and various intracellular compartments. Such interaction is dictated by individual elements in the carboxy terminal domain of the Ras proteins, including a farnesyl isoprenoid group, sequences in the hypervariable region (hvr)-linker, and palmitoyl groups in H/N Ras isoforms. 2. The farnesyl group acts as a specific recognition unit that interacts with prenyl-binding pockets in galectin-1 (Gal-1), galectin-3 (Gal-3), and cGMP phosphodiesterase delta. This interaction appears to contribute to the prolongation of Ras signals in the PM, the determination of Ras effector usage, and perhaps also the transport of cytoplasmic Ras. Gal-1 promotes H-Ras signaling to Raf at the expense of phosphoinositide 3-kinase (PI3-K) and Ral guanine nucleotide exchange factor (RalGEF), while galectin-3 promotes K-Ras signaling to both Raf and PI3-K. 3. The hvr-linker and the palmitates of H-Ras and N-Ras determine the micro- and macro-localizations of these proteins in the PM and in the Golgi, as well as in 'rasosomes', randomly moving nanoparticles that carry palmitoylated Ras proteins and their signal through the cytoplasm.4. The dynamic compartmentalization of Ras proteins contributes to the spatial organization of Ras signaling, promotes redistribution of Ras, and provides an additional level of selectivity to the signal output of this regulatory GTPase. PMID- 16691443 TI - Effect of ischemia in vivo and oxygen-glucose deprivation in vitro on NOS pools in the spinal cord: comparative study. AB - 1. This study was performed to compare both the Ca(2+)-dependent nitric oxide synthase (NOS) activity and the neuronal nitric oxide synthase immunoreactivity (nNOS-IR) in the rabbit lumbosacral spinal cord after 15 min abdominal aorta occlusion (ischemia in vivo) and oxygen-glucose deprivation of the spinal cord slices for 45 and 60 min (ischemia in vitro). All ischemic periods were followed by 15, 30 and 60 min reoxygenation in vitro. 2. Catalytic nitric oxide synthase activity was determined by the conversion of (L)-[(14)C]arginine to (L) [(14)C]citrulline. Neuronal nitric oxide synthase immunoreactivity in the spinal cord was detected by incubation of sections with polyclonal sheep-nNOS-primary antibody and biotinylated anti-sheep secondary antibody. 3. Our results show that ischemia in vivo and the oxygen-glucose deprivation of spinal cord slices in vitro result in a time-dependent loss of constitutive NOS activity with a partial restoration of enzyme activity during 15 and 45 min ischemia followed by 30 min of reoxygenation. A significant decrease of enzyme activity was found during 60 min ischemia alone, which persisted up to 1 h of oxygen-glucose restoration. The upregulation of neuronal nitric oxide synthase was observed in the ventral horn motoneurons after all ischemic periods. The remarkable changes in optical density of neuronal nitric oxide synthase immunoreactive motoneurons were observed after 45 and 60 min ischemia in vitro followed by 30 and 60 min reoxygenation. 4. Our results suggest that the oxygen-glucose deprivation followed by reoxygenation in the spinal cord is adequately sensitive to monitor ischemia/reperfusion changes. It seems that 15 min ischemia in vivo and 45 min ischemia in vitro cause reversible changes, while the decline of Ca(2+)-dependent nitric oxide synthase activity after 60 min ischemic insult suggests irreversible alterations. PMID- 16691449 TI - Effect of endometrial cavity fluid on clinical pregnancy rate in tubal embryo transfer (TET). AB - PURPOSE: In this study, we assess whether the endometrial cavity fluid (ECF) generated physiologically by the genital tract have negative effect on the pregnancy rate during tubal embryo transfer (TET) in patients who do not have hydrosalpinx or bilateral tubal obstruction. METHODS: We retrospectively collected samples from 176 women with 195 cycles receiving TET due to male factor, unexplained infertility or endometriosis from June 1999 to Dec. 2003, and divided them into two groups (group I: patient with fluid accumulation >1 mm in the anterior-posterior diameter in the uterine cavity; group II: patient without fluid accumulation in the uterine cavity). Endometrium thickness was measured as a maximal distance between anterior and posterior myometrium-endometrium interface under the long-axis view. The A-P diameter of ECF was measured via vaginal sonar on the day of ovum pick-up (OPU). RESULTS: From a total of 195 ART cycles involving 176 patients, the accumulation of ECF was detected in 10 cycles (5.1%). Seven of ten cycles (70%) with the accumulation of ECF were proved to be pregnant clinically. However, in the rest 185 cycles (group II), 98 of them (53%) were proved to be pregnant. At the mean time, the implantation rate was 29.03% and 25.71% in the groups I and II, respectively. No significant difference of the clinical pregnancy rate and the implantation rate was found between the two groups. CONCLUSIONS: According to our study, if the ECF was generated physiologically by the genital tract during controlled ovarian hyperstimulation (COH), the clinical pregnancy rate is not worse and no influence of embryo implantation was found. PMID- 16691456 TI - Investigation of a fluorescence signal amplification mechanism used for the direct molecular detection of nucleic acids. AB - A fluorescence signal amplification mechanism allowing detection limits for DNA in the zeptomolar range was investigated. Photophysical properties of the molecular system were studied in order to better explain the signal amplification that is observed. We show that the confinement of a fluorescent DNA hybridization transducer in aggregates improves its quantum yield and photostability. Furthermore, we show that the combination of the resonance energy transfer occurring within the aggregates with the use of a conjugated polymer as the hybridization transducer and donor allows ultrafast and efficient energy coupling to the aggregates and can lead to the excitation of a large number of acceptors by only one donor. PMID- 16691457 TI - Bias and accuracy of children's perceptions of peer acceptance: prospective associations with depressive symptoms. AB - Are depressive symptoms in middle childhood associated with more or less realistic social self-perceptions? At the beginning and end of the school year, children in grades 3 through 5 (n=667) rated how much they liked their classmates, predicted the acceptance ratings they would receive from each of their classmates, and completed self-report measures of perceived acceptance and depressive symptoms. Accuracy of perceived acceptance was indexed by the mean difference between pairs of predicted and received ratings (absolute values). Standardized residual scores created by regressing self-reported perceived acceptance (either predicted ratings or children's responses to a questionnaire measure of perceived peer acceptance) onto peer acceptance ratings formed two measures of bias. Bi-directional associations were found for accuracy of perceived acceptance and depressive symptoms; inaccurate perceptions predicted increases in depressive symptoms and depressive symptoms predicted decreased accuracy. Neither measure of bias predicted changes in depressive symptoms. Depressive symptoms predicted increases in negatively biased perceptions as assessed via questionnaire. PMID- 16691458 TI - Medical care of employees long-term sick listed due to mental health problems: a cohort study to describe and compare the care of the occupational physician and the general practitioner. AB - OBJECTIVES: To describe medical management by the general practitioner (GP) and occupational physician (OP) of workers sick listed due to mental health problems, and to determine agreement in diagnosis, main cause of sickness absence and obstacles in return to work. METHODS: A cohort of 555 employees being sick listed for 12 to 20 weeks due to mental health problems was recruited and followed for 1 year. These employees were interviewed about their mental health and contacts with GP, OP, other specialists and employer. In addition, the GP and/or the OP of 72 employees were interviewed about the medical diagnosis and management. RESULTS: Most employees sick listed for 12-20 weeks visited their GP and OP. According to the employees most interventions applied by the GP were medical interventions, such as referral of employees and prescription of medical drugs. Working conditions were seldom discussed by the GP and work-related interventions were never applied. Most interventions applied by the OP were work-related interventions and/or contact with the employer. The OP more often talked about working conditions and conflicts. According to the employees, the communication between GP and OP only took place in 8% of the cases. Agreement in the diagnosis, main cause of sickness absence, and obstacles in return to work reported by the GPs and OPs of the same employee was poor. In addition, similarity in reported diagnosis by GP and/or OP and the employees' scores on valid questionnaires on (mental) health was limited. CONCLUSIONS: The lack of communication and agreement by Dutch GPs and OPs in medical diagnosis and management of employees long-term sick listed due to mental health problems are indicators of sub-optimal medical treatment and return-to-work strategies. PMID- 16691461 TI - Association between cannabis use and sexual risk behavior among young heterosexual adults. AB - To study the association between cannabis use and frequent sexual risk behavior, we tested the hypothesis of a situational influence of cannabis use in sexual encounters using a combination of global association study and event-level analysis and examined possible mediator variables, including the personality trait of hedonism/risk preference, psychosocial stress, and HIV-related beliefs, using mediation models. The results of a computer-assisted telephone interview of a random sample of 2790 heterosexual men and women aged 16-24 years showed that risky sexual behavior was more frequent in cannabis-using men and women than in non-using persons. The results did not support a situational effect of cannabis intoxication on sexual risk behavior. The more frequent sexual risk behavior among cannabis users was mediated by decreased intentions to use HIV protection, by lower HIV-self-efficacy, and higher risk preference/hedonism. Only among women psychosocial stress was a partial mediator. The findings show that HIV prevention programs for cannabis-using young adults should emphasize the role of person variables instead of situation variables. PMID- 16691462 TI - Self-identification as "down low" among men who have sex with men (MSM) from 12 US cities. AB - Men who have sex with men (MSM) who are on the "down low" (DL) have been the subject of considerable media attention, but few data on this population exist. This exploratory study (N=455) compared MSM who considered themselves to be on the DL with MSM who did not (non-DL MSM). 20% self-identified as DL. Blacks and Hispanics were more likely than Whites to self identify as DL. MSM who did not identify as gay were more likely than gay-identified MSM to describe themselves as DL. DL-identified MSM were less likely to have had seven or more male partners in the prior 30 days, but were more likely to have had a female sex partner and to have had unprotected vaginal sex. DL-identified MSM were less likely to have ever been tested for HIV than were non-DL MSM. Prevention agencies should expand existing programs for MSM to include specific efforts to reach DL MSM. PMID- 16691463 TI - Are there distinctive trajectory groups in substance abuse remission over 10 years? An application of the group-based modeling approach. AB - This paper introduces to mental health services and evaluation researchers a method for analyzing longitudinal data: a group-based modeling approach for applied research. We present the basic formulations of the model and illustrate its application by analyzing the 10-year longitudinal data on substance abuse remission from the New Hampshire Dual Disorders Study. The basic features of the approach are: identifying latent groups with distinctive trajectories, estimating the proportion of the population in each group, linking individual-level covariates to group membership, assigning individuals to different groups, and creating group profiles based on posterior probabilities. We also discuss cautions and the controversies surrounding this approach. The major findings include four groups with distinct trajectories of remission from substance abuse. PMID- 16691464 TI - Transport ATPases: structure, motors, mechanism and medicine: a brief overview. AB - Today we know there are four different types of ATPases that operate within biological membranes with the purpose of moving many different types of ions or molecules across these membranes. Some of these ions or molecules are transported into cells, some out of cells, and some in or out of organelles within cells. These ATPases span the biological world from bacteria to eukaryotic cells and have become most simply and commonly known as "transport ATPases." The price that each cell type pays for transport work is counted in molecules of hydrolyzed ATP, a metabolic currency that is itself regenerated by a transport ATPase working in reverse, i.e., the ATP synthase. Four major classes of transport ATPases, the P, V, F, and ABC types are now known. In addition to being involved in many different types of biological/physiological processes, mutations in these proteins also account for a large number of diseases. The purpose of this introductory article to a mini-review series on transport ATPases is to provide the reader with a very brief and focused look at this important area of research that has an interesting history and bears significance to cell physiology, biochemistry, immunology, nanotechnology, and medicine, including drug discovery. The latter involves potential applications to a whole host of diseases ranging from cancer to those that affect bones (osteoporosis), ears (hearing), eyes (macromolecular degeneration), the heart (hypercholesterolemia/cardiac arrest,), immune system (immune deficiency disease), kidney (nephrotoxicity), lungs (cystic fibrosis), pancreas (diabetes and cystic fibrosis), skin (Darier disease), and stomach (ulcers). PMID- 16691465 TI - The structural basis for coupling of Ca2+ transport to ATP hydrolysis by the sarcoplasmic reticulum Ca2+-ATPase. AB - Recently, a series of structure determinations has nearly completed a structural description of the transport cycle of the sarcoplasmic reticulum Ca(2+)-ATPase, especially those steps concerned with the phosphorylation by ATP and the dephosphorylation reaction. From these structures Ca(2+)-ATPase emerges as a molecular machine, where globular cytosolic domains and transmembrane helices work in concert like a mechanical pump, as can be vividly demonstrated in animated versions of the pump cycle. The structures show that both ATP phosphorylation and dephosphorylation at Asp351 take place as nucleophilic SN2 reactions, which are associated with Ca(2+) and H(+) occluded states, respectively. These transitory steps ensure efficient coupling between Ca(2+) transport and ATP hydrolysis. PMID- 16691466 TI - Studies of Ca2+ ATPase (SERCA) inhibition. AB - The Ca(2+) transport ATPase of intracellular membranes (SERCA) can be inhibited by a series of chemical compounds such as Thapsigargin (TG), 2,5-di(tert butyl)hydroquinone (DBHQ) and 1,3-dibromo-2,4,6-tris (methyl-isothio-uronium) benzene (Br(2)-TITU). These compounds have specific binding sites in the ATPase protein, and different mechanisms of inhibition. On the other hand, SERCA gene silencing offers a convenient and specific method for suppression of SERCA activity in cells. The physiological and pharmacological implications of SERCA inhibition are discussed. PMID- 16691467 TI - Regulation of plant plasma membrane H+- and Ca2+-ATPases by terminal domains. AB - In the last few years, major progress has been made to elucidate the structure, function, and regulation of P-type plasma membrane H(+)-and Ca(2+)-ATPases. Even though a number of regulatory proteins have been identified, many pieces are still lacking in order to understand the complete regulatory mechanisms of these pumps. In plant plasma membrane H(+)- and Ca(2+)-ATPases, autoinhibitory domains are situated in the C- and N-terminal domains, respectively. A model for a common mechanism of autoinhibition is discussed. PMID- 16691468 TI - SERCA2a in heart failure: role and therapeutic prospects. AB - Ca(2+) is a key molecule controlling several cellular processes, from fertilization to cell death, in all cell types. In excitable and contracting cells, such as cardiac myocytes, Ca(2+) controls muscle contractility. The spatial and temporal segregation of Ca(2+) concentrations are central to maintain its concentration gradients across the cells and the cellular compartments for proper function. SERCA2a is a cornerstone molecule for maintaining a balanced concentration of Ca(2+) during the cardiac cycle, since it controls the transport of Ca(2+) to the sarcoplasmic reticulum (SR) during relaxation. Alterations of the activity of this pump have been widely investigated, emphasizing its central role in the control of Ca(2+) homeostasis and consequently in the pathogenesis of the contractile defect seen with heart failure. This review focuses on the molecular characteristics of the pump, its role during the cardiac cycle and the prospects derived from the manipulation of SERCA2a for heart failure treatment. PMID- 16691469 TI - Splice variants of the gamma subunit (FXYD2) and their significance in regulation of the Na, K-ATPase in kidney. AB - The recent discovery of a family of single-span membrane proteins (the FXYD proteins) introduced a new direction to the rather complicated area of regulation of Na, K-ATPase. At least six members of the family have been shown to associate with the Na, K-ATPase in a cell- and tissue-specific manner, while four of them, namely the gamma subunit (FXYD2), CHIF (FXYD4), phospholemman (FXYD1), and dysadherin (FXYD5) have been identified in kidney. All four exhibited different effects on the properties of the pump in heterologous expression systems. Taken along with their non-overlapping expression patterns in the nephron, this provides a potential structural basis for the segment-specific properties of the Na, K-ATPase that had been reported in a number of papers on kidney physiology. This brief review summarizes our own contributions on structure/functional characterization of one of the family members, the gamma subunit (FXYD2). The focus is on splice variants of gamma, their structural similarity and yet distinct effects conferred to Na, K-ATPase. PMID- 16691470 TI - Function of FXYD proteins, regulators of Na, K-ATPase. AB - In this short review, we summarize our work on the role of members of the FXYD protein family as tissue-specific modulators of Na, K-ATPase. FXYD1 or phospholemman, mainly expressed in heart and skeletal muscle increases the apparent affinity for intracellular Na(+) of Na, K-ATPase and may thus be important for appropriate muscle contractility. FXYD2 or gamma subunit and FXYD4 or CHIF modulate the apparent affinity for Na(+) of Na, K-ATPase in an opposite way, adapted to the physiological needs of Na(+) reabsorption in different segments of the renal tubule. FXYD3 expressed in stomach, colon, and numerous tumors also modulates the transport properties of Na, K-ATPase but it has a lower specificity of association than other FXYD proteins and an unusual membrane topology. Finally, FXYD7 is exclusively expressed in the brain and decreases the apparent affinity for extracellular K(+), which may be essential for proper neuronal excitability. PMID- 16691471 TI - Structure and regulation of the V-ATPases. AB - The V-ATPases are ATP-dependent proton pumps present in both intracellular compartments and the plasma membrane. They function in such processes as membrane traffic, protein degradation, renal acidification, bone resorption and tumor metastasis. The V-ATPases are composed of a peripheral V(1) domain responsible for ATP hydrolysis and an integral V(0) domain that carries out proton transport. Our recent work has focused on structural analysis of the V-ATPase complex using both cysteine-mediated cross-linking and electron microscopy. For cross-linking studies, unique cysteine residues were introduced into structurally defined sites within the B and C subunits and used as points of attachment for the photoactivated cross-linking reagent MBP. Disulfide mediated cross-linking has also been used to define helical contact surfaces between subunits within the integral V(0) domain. With respect to regulation of V-ATPase activity, we have investigated the role that intracellular environment, luminal pH and a unique domain of the catalytic A subunit play in controlling reversible dissociation in vivo. PMID- 16691472 TI - Close-up and genomic views of the yeast vacuolar H+-ATPase. AB - The yeast V-ATPase has emerged as an excellent model for other eukaryotic V ATPases. In this review, recent biochemical and genomic studies of the yeast V ATPase are described, with a focus on: 1) the role of V(1) subunit H in coupling ATP hydrolysis and proton pumping and 2) identification of the full set of yeast haploid deletion mutants that exhibit the pH and calcium-sensitive growth characteristic of loss of V-ATPase activity. The combination of "close-up" biochemical views of V-ATPase structure and mechanism and "geomic" views of its functional reach promises to provide new insights into the physiological of V ATPases. PMID- 16691473 TI - Rotation, structure, and classification of prokaryotic V-ATPase. AB - The prokaryotic V-type ATPase/synthases (prokaryotic V-ATPases) have simpler subunit compositions than eukaryotic V-ATPases, and thus are useful subjects for studying chemical, physical and structural properties of V-ATPase. In this review, we focus on the results of recent studies on the structure/function relationships in the V-ATPase from the eubacterium Thermus thermophilus. First, we describe single-molecule analyses of T. thermophilus V-ATPase. Using the single-molecule technique, it was established that the V-ATPase is a rotary motor. Second, we discuss arrangement of subunits in V-ATPase. Third, the crystal structure of the C-subunit (homolog of eukaryotic d-subunit) is described. This funnel-shape subunit appears to cap the proteolipid ring in the V(0) domain in order to accommodate the V(1) central stalk. This structure seems essential for the regulatory reversible association/dissociation of the V(1) and the V(0) domains. Last, we discuss classification of the V-ATPase family. We propose that the term prokaryotic V-ATPases should be used rather than the term archaeal-type ATPase (A-ATPase). PMID- 16691474 TI - Structure and mechanism of vacuolar Na+-translocating ATPase from Enterococcus hirae. AB - V-type Na(+)-ATPase from Entercoccus hirae consists of nine kinds of subunits (NtpA(3), B(3), C(1), D(1), E(1-3), F(1-3), G(1), I(1), and K(10)) which are encoded by the ntp operon. The amino acid sequences of the major subunits, A, B, and K (proteolipid), were highly similar to those of A, B, and c subunits of eukaryotic V-ATPases, and those of beta, alpha, and c subunits of F-ATPases. We modeled the A and B subunits by homology modeling using the structure of beta and alpha subunits of F-ATPase, and obtained an atomic structure of NtpK ring by X ray crystallography. Here we briefly summarize our current models of the whole structure and mechanism of the E. hirae V-ATPase. PMID- 16691475 TI - Selective assembly of V-ATPase subunit isoforms in mouse kidney. AB - The kidney plays vital roles in acid-base homeostasis, and the reabsorption of water, ions, and proteins. These processes are achieved through acidification of urine and endosomes of proximal tubule epithelial cells. Multisubunit vacuolar type proton ATPase (V-ATPase) is one of the major acidification-machinery proteins that localizes to the apical or basolateral plasma membranes of intercalated cells in collecting ducts and the endosomal region at the base of brush border microvilli in proximal tubules. Multiple subunit isoforms of V ATPase, which are expressed in kidney, have been identified. One obvious question is whether the pumps at different locations in the kidney have their own unique subunit identities. We have used a combination of methods to study this enzyme in kidney including immunocytochemical staining and immunoprecipitation analyses. The subunit isoforms of V-ATPase exhibited selective association/assembly in kidney: kidney-specific isoforms predominantly formed the intercalated cell proton pump, whereas the pump located in the brush border comprised ubiquitously expressed counterparts. PMID- 16691476 TI - Interactions between vacuolar H+-ATPases and microfilaments in osteoclasts. AB - Vacuolar H(+)-ATPases (V-ATPases) are transported from cytosolic compartments to the ruffled plasma membrane of osteoclasts as they activate to resorb bone. Transport of V-ATPases is essential for bone resorption, and is associated with binding interactions between V-ATPases and microfilaments that are mediated by an actin-binding site in subunit B. This site is contained within 44 amino acids in the amino terminal domain, and requires a sequence motif that resembles an actin binding motif found in mammalian profilin 1. Small alterations in the profilin like sequence disrupt the actin-binding activity of subunit B. The interaction between V-ATPases and microfilaments in osteoclasts is regulated in response to changes in phosphatidylinositol-3 kinase activity. During internalization of V ATPases from the plasma membrane of osteoclasts after a cycle of resorption, V ATPases bind microfilaments that are in podosomes, dynamic actin-based structures, also present in metastatic cancer cells. Studies are ongoing to establish the physiological role of the microfilament-binding activity of subunit B in osteoclasts and in other cells. PMID- 16691477 TI - Physiological and metabolic implications of V-ATPase isoforms in the kidney. AB - Not all vacuolar-type H(+)-ATPases (V-ATPases) are alike; those responsible for H(+) movement across plasma membranes contain some different, tissue-specific subunit isoforms. This brief review outlines those that have special relevance to the kidney, and illustrates their importance by describing various human diseases where loss of local proton pump function not only confers a severe phenotype, but has revealed related tissues where these same isoforms are expressed, signifying their physiological importance. PMID- 16691478 TI - V-ATPases as drug targets. AB - V-ATPases are large, complex enzymes responsible for acidification of many internal compartments in eukaryotic cells. They also occur on plasma membranes of specialized cells, where they acidify the surrounding milieu. Numerous physiological processes depend on the activity of V-ATPases, and V-ATPases are implicated as a contributing factor in multiple diseases, including osteoporosis, deafness, and cancer. Three classes of natural products have been identified as potent inhibitors of V-ATPases. The bafilomycins and concanamycins, which inhibit all known eukaryotic V-ATPases, are the most extensively studied inhibitors. They bind the Vo subunit c and may inhibit the enzyme by preventing rotation of the c subunit ring. The salicylihalamides and lobatamides show remarkable specificity for animal V-ATPases. The chondropsins preferentially inhibit the fungal V ATPase. Because of the variety of processes and diseases associated with V ATPases and the possibility of designing selective inhibitors, the V-ATPases are attractive targets for development of therapeutic agents. PMID- 16691479 TI - Identification of phosphate binding residues of Escherichia coli ATP synthase. AB - Four positively-charged residues, namely betaLys-155, betaArg-182, betaArg-246, and alphaArg-376 have been identified as Pi binding residues in Escherichia coli ATP synthase. They form a triangular Pi binding site in catalytic site betaE where substrate Pi initially binds for ATP synthesis in oxidative phosphorylation. Positive electrostatic charge in the vicinity of betaArg-246 is shown to be one important component of Pi binding. PMID- 16691480 TI - Zooming in on ATP hydrolysis in F1. AB - We summarize our current view of the reaction mechanism in F(1)-ATPase as it has emerged from experiment, theory, and computational studies over the last several years. ATP catalysis in the catalytic binding pockets of F(1) takes place without the release of any significant free energy and is efficiently driven by the combined action of two water molecules utilizing a so-called protein-relay mechanism. The chemical reaction itself is controlled by the spatial position of a key arginine residue. PMID- 16691481 TI - Structure and function of subunit a of the ATP synthase of Escherichia coli. AB - The structure of subunit a of the Escherichia coli ATP synthase has been probed by construction of more than one hundred monocysteine substitutions. Surface labeling with 3-N-maleimidyl-propionyl biocytin (MPB) has defined five transmembrane helices, the orientation of the protein in the membrane, and information about the relative exposure of the loops connecting these helices. Cross-linking studies using TFPAM-3 (N-(4-azido-2,3,5,6-tetrafluorobenzyl)-3 maleimido-propionamide) and benzophenone-4-maleimide have revealed which elements of subunit a are near subunits b and c. Use of a chemical protease reagent, 5-( bromoacetamido)-1,10-phenanthroline-copper, has indicated that the periplasmic end of transmembrane helix 5 is near that of transmembrane helix 2. PMID- 16691482 TI - Chemomechanical coupling in single-molecule F-type ATP synthase. AB - An extremely small reaction chamber with a volume of a few femtoliters was developed for a highly sensitive detection of biological reaction. By encapsulating a single F(1)-ATPase (F(1)) molecule with ADP and an inorganic phosphate in the chamber, the chemomechanical coupling efficiency of ATP synthesis catalyzed by reversely rotated F(1) was successfully determined (Rondelez et al., 2005a, Nature, 444, 773-777). While the alpha3beta3gamma subcomplex of F(1) generated ATP with a low efficiency (approximately 10%), inclusion of the epsilon subunit into the subcomplex enhanced the efficiency up to 77%. This raises a new question about the mechanism of F(0)F(1)-ATP synthase (F(0)F(1)): How does the epsilon subunit support the highly coupled ATP synthesis of F(1)? To address this question, we measured the conformational dynamics of the epsilon subunit using fluorescence resonance energy transfer (FRET) at the single molecule level. The experimental data revealed epsilon changes the conformation of its C-terminus helices in a nucleotide-dependent manner. It is plausible that the conformational change of epsilon switches the catalytic mode of F(0)F(1) for highly coupled ATP synthesis. PMID- 16691483 TI - ATP synthases with novel rotor subunits: new insights into structure, function and evolution of ATPases. AB - ATPases with unusual membrane-embedded rotor subunits were found in both F(1)F(0) and A(1)A(0) ATP synthases. The rotor subunit c of A(1)A(0) ATPases is, in most cases, similar to subunit c from F(0). Surprisingly, multiplied c subunits with four, six, or even 26 transmembrane spans have been found in some archaea and these multiplication events were sometimes accompanied by loss of the ion translocating group. Nevertheless, these enzymes are still active as ATP synthases. A duplicated c subunit with only one ion-translocating group was found along with "normal" F(0) c subunits in the Na(+) F(1)F(0) ATP synthase of the bacterium Acetobacterium woodii. These extraordinary features and exceptional structural and functional variability in the rotor of ATP synthases may have arisen as an adaptation to different cellular needs and the extreme physicochemical conditions in the early history of life. PMID- 16691484 TI - Ectopic localization of mitochondrial ATP synthase: a target for anti angiogenesis intervention? AB - A receptor for angiostatin was identified on the surface of endothelial cells as F(1)-F(0) ATP synthase (Moser et al., 1999). Proc. Natl. Acad. Sci. U.S.A. 96, 2811-2816. This ectopic ATP synthase catalyzes ATP synthesis and is inhibited by angiostatin over a wide pH range. Endothelial cells grown at normal pH suffer no ill effects from this angiostatin-mediated inhibition of ATP synthase, whereas endothelial cells grown at low, tumor-like extracellular pH cannot maintain a normal intracellular pH and die. Angiostatin inhibits both ATP synthesis and ATP hydrolysis (Moser et al., 2001) and interferes with intracellular pH regulation (Wahl and Grant, 2002; Wahl et al., 2002). Although angiostatin administered intravenously is cleared from the circulation in a matter of minutes, angiostatin mimetics that are more stable have potential for clinical application. An angiostatin-mimetic activity has recently been observed using a polyclonal antibody against the beta catalytic subunit of ATP synthase. In order to explore the mechanism of action of angiostatin and its mimetics, further work needs to be done to evaluate clinical applicability, specificity, and contraindications for this class of therapeutics. PMID- 16691485 TI - Coupling proton movement to ATP synthesis in the chloroplast ATP synthase. AB - The chloroplast F(0)F(1)-ATP synthase-ATPase is a tiny rotary motor responsible for coupling ATP synthesis and hydrolysis to the light-driven electrochemical proton gradient. Reversible oxidation/reduction of a dithiol, located within a special regulatory domain of the gamma subunit of the chloroplast F(1) enzyme, switches the enzyme between an inactive and an active state. This regulatory mechanism is unique to the ATP synthases of higher plants and its physiological significance lies in preventing nonproductive depletion of essential ATP pools in the dark. The three-dimensional structure of the chloroplast F(1) gamma subunit has not yet been solved. To examine the mechanism of dithiol regulation, a model of the chloroplast gamma subunit was obtained through segmental homology modeling based on the known structures of the mitochondrial and bacterial gamma subunits, together with de novo construction of the unknown regulatory domain. The model has provided considerable insight into how the dithiol might modulate catalytic function. This has, in turn, suggested a mechanism by which rotation of subunits in F(0), the transmembrane proton channel portion of the enzyme, can be coupled, via the epsilon subunit, to rotation of the gamma subunit of F(1) to achieve the 120 degrees (or 90 degrees +30 degrees) stepping action that is characteristic of F(1) gamma subunit rotation. PMID- 16691486 TI - Nucleotide binding domain interactions during the mechanochemical reaction cycle of ATP-binding cassette transporters. AB - ATP-binding cassette (ABC) transporters serve as importers and exporters for a wide variety of solutes in both prokaryotes and eukaryotes, and are implicated in microbial drug resistance and a number of significant human genetic disorders. Initial crystal structures of the soluble nucleotide binding domains (NBDs) of ABC transporters, while a significant step towards understanding the coupling of ATP binding and hydrolysis to transport, presented researchers with important questions surrounding the role of the signature sequence residues, the composition of the nucleotide binding sites, and the mode of NBD dimerization during the transport reaction cycle. Recent studies have begun to address these concerns. This mini-review summarizes the biochemical and structural characterizations of two archaebacterial NBDs from Methanocaldococcus jannaschii, MJ0796 and MJ1267, and offers current perspectives on the functional mechanism of ABC transporters. PMID- 16691487 TI - New insights into the drug binding, transport and lipid flippase activities of the p-glycoprotein multidrug transporter. AB - The MDR1 P-glycoprotein, an ATP-binding cassette (ABC) superfamily member that functions as an ATP-driven drug efflux pump, has been linked to resistance of human tumors to multiple chemotherapeutic agents. P-glycoprotein binds and actively transports a large variety of hydrophobic drugs and peptides. P glycoprotein in reconstituted proteoliposomes is also an outwardly directed flippase for membrane phospholipids and simple glycosphinglipids. This review focuses on recent advances in our understanding of P-glycoprotein structure and function, particularly through the use of fluorescence spectroscopic approaches. Progress is being made towards understanding the structure of the transporter, especially the spatial relationship between the two nucleotide-binding domains. Exploration of the P-glycoprotein catalytic cycle using vanadate-trapped complexes has revealed that drug transport likely takes place by concerted conformational changes linked to relaxation of a high energy intermediate. Low resolution mapping of the protein using fluorescence resonance energy transfer showed that both the H and R drug-binding sites are located within the cytoplasmic leaflet. Two drugs can bind to the R-site simultaneously, suggesting that the protein contains a large flexible binding region. PMID- 16691488 TI - The remarkable transport mechanism of P-glycoprotein: a multidrug transporter. AB - Human P-glycoprotein (ABCB1) is a primary multidrug transporter located in plasma membranes, that utilizes the energy of ATP hydrolysis to pump toxic xenobiotics out of cells. P-glycoprotein employs a most unusual molecular mechanism to perform this drug transport function. Here we review our work to elucidate the molecular mechanism of drug transport by P-glycoprotein. High level heterologous expression of human P-glycoprotein, in the yeast Saccharomyces cerevisiae, has facilitated biophysical studies in purified proteoliposome preparations. Development of novel spin-labeled transport substrates has allowed for quantitative and rigorous measurements of drug transport in real time by EPR spectroscopy. We have developed a new drug transport model of P-glycoprotein from the results of mutagenic, quantitative thermodynamic and kinetic studies. This model satisfactorily accounts for most of the unusual kinetic, coupling, and physiological features of P-glycoprotein. Additionally, an atomic detail structural model of P-glycoprotein has been devised to place our results within a proper structural context. PMID- 16691489 TI - The occluded nucleotide conformation of p-glycoprotein. AB - We review recent work on E552A/E1197A P-glycoprotein. This ATPase-defective mutant occludes MgATP tightly with maximal 1/1 stoichiometry in drug-sensitive fashion. The occluded nucleotide conformation appears to represent a transient, asymmetric, catalytic intermediate. We present a model for catalysis incorporating nucleotide binding domain (NBD) dimerization and the occluded nucleotide conformation, and we speculate as to how catalysis seen in P glycoprotein might be harmonized with symmetrical dimer structures of isolated NBDs. PMID- 16691490 TI - Rescue of folding defects in ABC transporters using pharmacological chaperones. AB - The ATP-binding cassette (ABC) family of membrane transport proteins is the largest class of transporters in humans (48 members). The majority of ABC transporters function at the cell surface. Therefore, defective folding and trafficking of the protein to the cell surface can lead to serious health problems. The classic example is cystic fibrosis (CF). In most CF patients, there is a deletion of Phe508 in the CFTR protein (DeltaF508 CFTR) that results in defective folding and intracellular retention of the protein (processing mutant). A potential treatment for most patients with CF would be to use a ligand(s) of CFTR that acts a pharmacological chaperone to correct the folding defect. The feasibility of such an approach was first demonstrated with the multidrug transporter P-glycoprotein (P-gp), an ABC transporter, and a sister protein of CFTR. It was found that P-gps with mutations at sites equivalent to those found in CFTR processing mutants were rescued when they were expressed in the presence of drug substrates or modulators of P-gp. These compounds acted as pharmacological chaperones and functioned by promoting interactions among the various domains in the protein during the folding process. Several groups have attempted to identify compounds that could rescue the folding defect in DeltaF508 CFTR. The best compound identified through high-throughout screening is a quinazoline derivative (CFcor-325). Expression of DeltaF508 CFTR as well as other CFTR processing mutants in the presence of 1 muM CFcor-325 promoted folding and trafficking of the mutant proteins to the cell surface in an active conformation. Therefore, CFcor-325 and other quinazoline derivates could be important therapeutic compounds for the treatment of CF. PMID- 16691491 TI - The intracellular antigen transport machinery TAP in adaptive immunity and virus escape mechanisms. AB - The transporter associated with antigen processing (TAP) is a crucial element of the adaptive immune system, which translocates proteasomal degradation products into the endoplasmic reticulum, for transfer of these peptides on major histocompatibility complex (MHC) I molecules within a macromolecular peptide loading complex. After loading and intracellular transport to the cell surface, these peptide/MHC complexes are monitored by cytotoxic T-lymphocytes. This review summarizes the structural organization and function of the ABC transporter TAP. Furthermore, we discuss human diseases and viral evasion strategies associated with TAP function. PMID- 16691492 TI - Loss of ubiquitin binding is a unifying mechanism by which mutations of SQSTM1 cause Paget's disease of bone. AB - Ubiquitin-associated (UBA) domain mutations of SQSTM1 are an important cause of Paget's disease of bone (PDB), which is a human skeletal disorder characterized by abnormal bone turnover. We previously showed that, when introduced into the full-length SQSTM1 protein, the disease-causing P392L, M404V, G411S, and G425R missense mutations and the E396X truncating mutation (representative of all of the SQSTM1 truncating mutations) cause a generalized loss of monoubiquitin binding and impaired K48-linked polyubiquitin binding at physiological temperature. Here, we show that the remaining three known PDB missense mutations, P387L, S399P, and M404T, have similar deleterious effects on monoubiquitin binding and K48-linked polyubiquitin binding by SQSTM1. The P387L mutation affects an apparently unstructured region at the N terminus of the UBA domain, some five residues from the start of the first helix, which is dispensable for polyubiquitin binding by the isolated UBA domain. Our findings support the proposal that the disease mechanism in PDB with SQSTM1 mutations involves a common loss of ubiquitin binding function of SQSTM1 and implicate a sequence extrinsic to the compact globular region of the UBA domain as a critical determinant of ubiquitin recognition by the full-length SQSTM1 protein. PMID- 16691493 TI - Ameloblastin fusion protein enhances pulpal healing and dentin formation in porcine teeth. AB - Ameloblastin (Ambn, also named "amelin" or "sheathlin") is a protein participating in enamel formation and mesenchymal-ectodermal interaction during early dentin formation in developing teeth. Experiments have demonstrated an association between Ambn expression and healing of acute pulp wounds. The purpose of this study was to investigate if local application of recombinant fusion Ambn (rAmbn) could influence reparative dentin formation in pulpotomized teeth. In this randomized, double-blinded study, pulpotomy was performed in 28 lower central incisors in 17 adult miniature pigs. Following the surgical procedure, the exposed pulp tissue was covered either with rAmbn or with calcium hydroxide. After 2, 4, or 8 weeks, the teeth were extracted and examined by histomorphometry and immunohistochemistry using antibodies against porcine ameloblastin, collagen type I, and dentin sialoprotein (DSP). In rAmbn-treated teeth, a substantial amount of newly formed reparative dentin was observed at the application site, completely bridging the pulpal wound. Dentin formation was also observed in calcium hydroxide-treated teeth; however, the amount of reparative dentin was significantly smaller (P < 0.001) than after rAmbn treatment. Immunohistochemistry confirmed that the new hard tissue formed was similar to dentin. This is the first time a direct link between ameloblastin and dentin formation has been made in vivo. The results suggest potential for rAmbn as a biologically active pulp-dressing agent for enhanced pulpal wound healing and reparative dentin formation after pulpotomy procedures. PMID- 16691494 TI - Tissue inhibitor of matrix metalloproteinase-1 suppresses apoptosis of mouse bone marrow stromal cell line MBA-1. AB - We investigated the action of tissue inhibitor of metalloproteinase-1 (TIMP-1) on apoptosis and differentiation of mouse bone marrow stromal cell line MBA-1. TIMP 1 did not affect alkaline phosphatase (ALP) activity, suggesting that it is not involved in osteoblastic differentiation in MBA-1 cells. However, TIMP-1 inhibited MBA-1 apoptosis induced by serum deprivation in a dose-dependent manner. Our study also showed increased Bcl-2 protein expression and decreased Bax protein expression with TIMP-1 treatment. TIMP-1 decreased cytochrome c release and caspase-3 activation in MBA-1 cells. TIMP-1 activated phosphatidylinositol 3-kinase (PI3-kinase) and c-Jun N-terminal kinase (JNK), and the PI3-kinase inhibitor LY294002 or the JNK inhibitor SP600125 abolished its antiapoptotic activity. To investigate whether antiapoptotic action of TIMP-1 was mediated through its inhibition on MMP activities, we constructed mutant TIMP-1 by side-directed mutagenesis, which abolished the inhibitory activity of MMPs by deletion of Cys1 to Ala4. Wild-type TIMP-1 and mutant TIMP-1 expression plasmids were transfected in MBA-1 cells, and results showed that mutant TIMP-1 still protected the induced MBA-1 cell against apoptosis. These data suggest that TIMP 1 antiapoptotic actions are mediated via the PI3-kinase and JNK signaling pathways and independent of TIMP-1 inhibition of MMP activities. PMID- 16691495 TI - Effects of radiation therapy on chondrocytes in vitro. AB - The negative irradiation complications of growth loss leading to limb length asymmetry and pathological fracture incurred following radiation therapy in pediatric patients has led to a renewed interest in understanding the specific effects of irradiation on the growth plate and the surrounding bone. In the present report, we examined the radiation therapy effects on primary rat growth cartilage chondrocytes in order to determine the chondrocyte radiosensitivity relative to other bone cell constituents and tumor cells, the postirradiation temporal progression of radiation-induced alterations in chondrocyte function, and the time course for the functional restoration of chondrocyte pathways that drive the eventual recovery in growth function. We employed an in vitro primary rat costochondral growth cartilage cell culture model system to evaluate the radiation therapy effects on proliferative chondrocytes using serial radiation doses (0-20 Gy) that are well within the clinically relevant range. Following irradiation, all of the following occurred in a dose-dependent manner: proliferation decreased, cytotoxicity increased, several markers of apoptosis increased, markers of radiation-induced cellular differentiation increased, and cell synthetic activity was disturbed. Alterations in proliferation, cell death, and induction of apoptosis are likely due to a transient radiation-induced derangement of the parathyroid hormone-related protein-Indian hedgehog proliferation-maturation pathway. Alterations in cellular differentiation and cell synthetic activity are novel observations for chondrocytes. Further, these results correspond very well to our previous work in an in vivo Sprague-Dawley rat model, making this model particularly relevant to researching the radiation therapy effects on longitudinal growth. PMID- 16691496 TI - A serum factor that recalcifies demineralized bone is conserved in bony fish and sharks but is not found in invertebrates. AB - We investigated the evolutionary origin of a serum activity that induces calcification within a type I collagen matrix, an activity previously described in rat and bovine serum. Serum was obtained from vertebrates with calcified tissues (bony fish and shark), vertebrates without calcified tissues (lamprey and hagfish), and three invertebrates (marine worm, crab, and sea urchin). Serum from the bony fish and shark proved to contain a potent nucleator of collagen calcification; like the previously described calcifying activity in rat serum, the fish and shark activities are both able to recalcify a demineralized rat tibia when tested in Dulbecco's modified Eagle medium containing as little as 1.5% of the respective serum and have an apparent molecular weight of 50-150 kDa. No calcifying activity could be detected in any of several experimental tests of invertebrate or hagfish serum. Weak calcifying activity could be detected in lamprey serum, but calcification was restricted to the growth plate of the decalcified tibia, with no detectable calcification in the type I collagen of the midshaft. These studies reveal a correlation between the evolutionary timing of the appearance of calcified tissues in vertebrates and the appearance of the serum activity that initiates calcification within collagen and, therefore, support the hypothesis that this serum activity may play a role in normal calcification of bone. PMID- 16691498 TI - Lactosylceramide is required in apoptosis induced by N-Smase. AB - Lactosylceramide (LacCer) is a member of the glycosphingolipid family which has been recently recognized as a signaling intermediate in the regulation of cell proliferation and cell adhesion. In this paper, we present our studies pointing to a potential role of LacCer in inducing apoptosis. In our studies we employed a human osteosarcoma cell line MG-63 (wild type, WT) and a neutral sphingomyelinase (N-SMase) deficient cell line CC derived from MG-63 (mutant) cells. We observed that WT cells were highly sensitive to tumor necrosis factor-alpha (TNF-alpha), ceramide and LacCer-induced apoptosis. In contrast, the mutant cells were insensitive to TNF-alpha-induced apoptosis as they did not generate ceramide and LacCer. However, the exogenous supply of ceramide and/or LacCer rendered the mutant cells apoptotic. Interestingly, preincubation of cells with D-threo-1 phenyl-2-decanoylamino-3-morpholino-1-propanol (D-PDMP), an inhibitor of glucosylceramide synthase and lactosylceramide synthase, abrogated ceramide induced apoptosis but not LacCer-induced apoptosis in both WT cells and the mutant cells. Moreover, TNF-alpha and LacCer-induced apoptosis required the generation of reactive oxygen species (ROS) in WT cells. However, since mutant cells did not produce significant amounts of LacCer and ROS in response to TNF alpha treatment they are insensitive to TNF-alpha-induced apoptosis. In summary, our studies suggest that TNF-alpha-induced N-SMase activation and production of ceramide is required to activate the apoptosis pathway in human osteosarcoma cells. But it is not sufficient to induce apoptosis. Rather, the conversion of ceramide to LacCer and ROS generation are critical for apoptosis. PMID- 16691499 TI - Efflux of sphingolipids metabolically labeled with [1-3H]sphingosine, L-[3- 3H]serine and [9,10- 3H]palmitic acid from normal cells in culture. AB - The membrane complex lipids of human fibroblasts and differentiated rat cerebellar granule cells in culture were metabolically radiolabeled with [1 (3)H]sphingosine, L-[3-(3)H]serine and [9,10-(3)H]palmitic acid. A relevant efflux of radioactive sphingolipids and phosphatidylcholine was observed from cells to the culture medium in the presence of fetal calf serum. This event was independent of the concentration and structure of the metabolic precursor administered to cells, and it was linearly time-dependent. The radioactive lipid patterns present in the medium were different from those present in the cells. Radioactive sphingomyelin and ganglioside GM3 containing short acyl chains were the main species present in the medium from human fibroblasts, while sphingomyelin and GD3 ganglioside in that from neuronal cells. In the absence of proteins in the culture medium, the efflux of complex lipids was much lower than in the presence of serum, and the patterns of released molecules were again different from those of cells. PMID- 16691500 TI - Altered expression and glycosylation of plasma proteins in rheumatoid arthritis. AB - Altered glycosylation of plasma proteins has been directly implicated in the pathogenesis of rheumatoid arthritis (RA). The present study investigated the changes in the Concanavalin-A (Con-A)-bound plasma proteins in the RA patients in comparison to that of the healthy controls. Two proteins (MW approximately 32 kDa and approximately 62 kDa) showed an alteration in expression while an altered monosaccharide profile (high mannose) was observed in the approximately 62 kDa protein in the samples collected from RA patients. The 2-dimensional polyacrylamide gel electrophoresis analysis of the Con-A-bound plasma samples showed a large number of protein spots, a few of which were differentially expressed in the RA patients. Some unidentified proteins were detected in the RA patients which were absent in the control samples. The present study, therefore, enunciates the role of carbohydrates as well as that of the acute phase response in the disease pathogenesis. PMID- 16691501 TI - Apoptosis of human breast carcinoma cells in the presence of cis-platin and L-/D PPMP: IV. Modulation of replication complexes and glycolipid: Glycosyltransferases. AB - Apoptosis of human breast carcinoma cells (SKBR-3, MCF-7, and MDA-468) has been observed after treatment of these cells with anti-cancer drug cis-platin and glycosphingolipid biosynthesis inhibitor L- and D-PPMP, respectively. These drugs initiated apoptosis in a dose-dependent manner as measured by phenotypic morphological changes, by binding of a fluorescent phophatidyl serine-specific dye (PSS-380) onto the outer leaflet of the cell membranes, and by activation of caspases, -3, -8, and -9. It was observed that in two hours very little apoptotic process had started but predominant biochemical changes occurred after 6 h. DNA degradation started after 24 hours of drug treatment. However, very little is known about the stability of the ';Replication Complexes'' during the apoptotic process. DNA helicases are motor proteins that catalyze the melting of genomic DNA during its replication, repair, and recombination processes. Previously, DNA helicase-III was characterized as a component of the replication complexes isolated from embryonic chicken brains as well as breast and colon carcinoma cells. Helicase activities were measured by a novel method (ROME assay), and DNA polymerase-alpha activities were determined by regular chain extension of the nicked ACT-DNA, by determining values obtained from +/- aphidicolin-treated incubation mixtures. In all three breast carcinoma cell lines, a common trend was observed: a decrease of activities of DNA polymerase-alpha and Helicase III. A sharp decrease of activities of the glycolipid sialyltransferases: SAT-2 (CMP NeuAc; GD3 alpha2-8 sialyltransferase) and SAT-4 (CMP-NeuAc: GM1a alpha2-3 sialyltransferase) was observed in the apoptotic carcinoma cells treated with L PPMP compared with cis-platin. PMID- 16691502 TI - Sphingolipids, cholesterol, and HIV-1: a paradigm in viral fusion. AB - Our previous studies show that the depletion of cholesterol or sphingolipids (raft-associated lipids) from receptor-bearing adherent cell lines blocks HIV-1 entry and HIV-1 Env-mediated membrane fusion. Here we have evaluated the mechanism(s) by which these lipids contribute to the HIV-1 Env-mediated membrane fusion. We report the following: (1) GSL depletion from a suspension T lymphocyte cell line (Sup-T1) reduced subsequent fusion with HIV-1IIIB-expressing cells by 70%. (2) Cholesterol depletion from NIH3T3 cells bearing HIV-1 receptors (NIH3T3CD4R5/NIH3T3CD4X4) did not impair subsequent fusion with HeLa cells expressing the corresponding HIV-1 Envs. In contrast GSL depletion from these targets reduced fusion by 50% suggesting that GSL facilitate fusion in different ways. (3) GSL-deficient GM95 cells bearing high receptors fused with HIV-1 Env expressing cells at 37 degrees C with kinetics similar to that of GSL + NIH3T3 targets. Based on these observations, we propose that the plasma membrane cholesterol is required to maintain the integrity of receptor pools whereas GSLs are involved in stabilizing the coupling of inter-receptor pools. PMID- 16691503 TI - Increased expression of peripheral benzodiazepine receptor (PBR) in dimethylbenz[a]anthracene-induced mammary tumors in rats. AB - Expression of peripheral benzodiazepine receptors (PBR) has been found in every tissue examined; however, it is most abundant in steroid-producing tissues. Although the primary function of PBR is the regulation of steroidogenesis, its existence in nonsteroidogenic tissues as well as in other cellular compartments including the nucleus suggests that there may be other roles for PBR. Our laboratory reported earlier a significant increase of PBR density in the nucleus of DMBA-induced malignant submandibular glands of rats, suggesting a role of PBR in nuclear events of peripheral tissues. Since then numerous studies have demonstrated the abundance of PBR in tumors. Numerous studies implicate a role for cholesterol in the mechanisms underlying cell proliferation and cancer progression. Based on studies with a battery of human breast cancer cell lines and several human tissue biopsies, Hardwick et al. suggested that PBR expression, nuclear localization, and PBR-mediated cholesterol transport into the nucleus are involved in human breast cancer cell proliferation and aggressive phenotype expression. The purpose of the present study is to confirm this hypothesis by developing an animal breast cancer model and correlating the above events with the breast cancer. Weanling rats were maintained on a diet containing animal protein (casein) for 30 days and then a single dose of DMBA in sesame oil (80 mg/kg) was administered by gavage to the animals. Control animals received the vehicle only. After 122 days of DMBA administration, the animals were sacrificed. All tumors were detected by palpation. B(max) of PBRs was 52.6% and 128.4% higher in the non-aggressive and aggressive cancer tissues, respectively, than that in normal tissues. Cholesterol uptake into isolated nuclei was found to be higher in both non-aggressive and aggressive tumor breast tissue than that in control tissue. There was also corresponding increase in B(max) of PBRs in the nucleus of cancer tissues. Furthermore, the nuclear nucleoside triphosphatase (NTPase) activity was found to be higher in aggressive tumor tissues than that in non aggressive tumor tissues. In conclusion, these data suggest that PBR ligand binding, and PBR-mediated cholesterol transport into the nucleus may be involved in the development of mammary gland adenocarcinoma, thus participating in the advancement of the disease. PMID- 16691504 TI - Potentiation of angiogenic switch in capillary endothelial cells by cAMP: A cross talk between up-regulated LLO biosynthesis and the HSP-70 expression. AB - During tumor growth and invasion, the endothelial cells from a relatively quiescent endothelium start proliferating. The exact mechanism of switching to a new angiogenic phenotype is currently unknown. We have examined the role of intracellular cAMP in this process. When a non-transformed capillary endothelial cell line was treated with 2 mM 8Br-cAMP, cell proliferation was enhanced by approximately 70%. Cellular morphology indicated enhanced mitosis after 32-40 h with almost one-half of the cell population in the S phase. Bcl-2 expression and caspase-3, -8, and -9 activity remained unaffected. A significant increase in the Glc(3)Man(9)GlcNAc(2)-PP-Dol biosynthesis and turnover, Factor VIIIC N glycosylation, and cell surface expression of N-glycans was observed in cells treated with 8Br-cAMP. Dol-P-Man synthase activity in the endoplasmic reticulum membranes also increased. A 1.4-1.6-fold increase in HSP-70 and HSP-90 expression was also observed in 8Br-cAMP treated cells. On the other hand, the expression of GRP-78/Bip was 2.3-fold higher compared to that of GRP-94 in control cells, but after 8Br-cAMP treatment for 32 h, it was reduced by 3-fold. GRP-78/Bip expression in untreated cells was 1.2-1.5-fold higher when compared with HSP-70 and HSP-90, whereas that of the GRP-94 was 1.5-1.8-fold lower. After 8Br-cAMP treatment, GRP-78/Bip expression was reduced 4.5-4.8-fold, but the GRP-94 was reduced by 1.5-1.6-fold only. Upon comparison, a 2.9-fold down-regulation of GRP 78/Bip was observed compared to GRP-94. We, therefore, conclude that a high level of Glc(3)Man(9)GlcNAc(2)-PP-Dol, resulting from 8Br-cAMP stimulation up-regulated HSP-70 expression and down-regulated that of the GRP-78/Bip, maintained adequate protein folding, and reduced endoplasmic reticulum stress. As a result capillary endothelial cell proliferation was induced. PMID- 16691505 TI - Nuclear factor inducing kinase: a key regulator in osteopontin- induced MAPK/IkappaB kinase dependent NF-kappaB-mediated promatrix metalloproteinase-9 activation. AB - Osteopontin (OPN) is a secreted, non-collagenous, sialic-acid rich, glycosylated adhesive phospho- protein. Several highly metastatic transformed cells synthesized a higher level of OPN compared with non-tumorigenic cells. We have recently reported that OPN induces nuclear factor-kappaB (NF-kappaB)-mediated promatrix metalloproteinase-2 activation through IkappaBalpha/IKK signaling pathways. However, the molecular mechanism(s) by which OPN regulates pro-matrix metalloproteinase-9 (pro-MMP-9) activation and involvement of upstream kinases in regulation of these processes that ultimately control cell motility and tumor growth in murine melanoma cells are not well defined. Here we report that OPN induces alphavbeta3 integrin-mediated phosphorylation and activation of nuclear factor inducing kinase (NIK) and enhances the interaction between phosphorylated NIK and IkappaBalpha kinase alpha/beta (IKKalpha/beta) in B16F10 cells. Moreover, NIK is involved in OPN-induced phosphorylations of MEK-1 and ERK1/2 in these cells. OPN induces NIK-dependent NF-kappaB activation through ERK/IKKalpha/beta mediated pathways. Furthermore, OPN enhances NIK-regulated urokinase-type plasminogen activator (uPA) secretion, uPA-dependent pro-MMP-9 activation, and cell motility. Pretreatment of cells with anti-MMP-2 antibody along with anti-MMP 9 antibody drastically inhibited the OPN-induced cell migration and chemoinvasion, whereas cells pretreated with anti-MMP-2 antibody had no effect on OPN-induced pro-MMP-9 activation suggesting that OPN induces pro-MMP-2 and pro MMP-9 activations through two distinct pathways. Taken together, NIK acts as crucial regulator in OPN-induced MAPK/IKK-mediated NF-kappaB-dependent uPA secretion and MMP-9 activation thereby controlling melanoma cell motility and chemoinvasion. PMID- 16691506 TI - Role of C-reactive protein in complement-mediated hemolysis in Malaria. AB - Human C-reactive protein (CRP) is a clinically important classical acute phase protein. Although CRP has been reported to bind with many nucleated cells, the direct binding of CRP to erythrocytes in diseases remains largely unexplored. The main focus of the present study was to investigate the binding of disease specific CRP to erythrocytes of same patients. Distinct molecular variant of disease-specific CRP was affinity purified from sera of malaria patients (CRP(Mal)). This CRP showed strong binding with malaria erythrocytes (RBC(Mal)) as confirmed by flow cytometric analysis (FACS), enzyme-linked immunosorbent assays (ELISA), and radio binding assays. Calcium and phosphoryl choline (PC) were found to be essential for this interaction. A 2.3-fold increased binding of induced CRP to RBC(Mal) as compared to normal erythrocytes (RBC(N)) confirmed disease-specificity. Preincubation of RBC(Mal) with unconjugated CRP showed 3-5 fold inhibition. The association constant of CRP and RBC(Mal) was 4.7 x 10(6) cpm/microg with the corresponding number of receptors/cell being 4.3 x 10(5). The effector function of CRP(Mal) has been demonstrated by its potency to activate the complement pathway. An optimal dose of 10 microg/ml of CRP induced three-fold higher hemolysis of patient erythrocytes as compared to RBC(N). These studies provide direct evidence for an important phagocytic functional interaction of this acute-phase protein by triggering the CRP-complement pathway after the binding of CRP(Mal) with RBC(Mal). Hemolysis as triggered by this pathway may be one of the causative factors of anemia, a common clinical manifestation of this disease. PMID- 16691507 TI - Structural studies on mannose-selective glycoprotein receptors using molecular modeling techniques. AB - Glycoproteins play important roles in various cellular events and their presence in appropriate locations in proper active conformations is essential for many biochemical functions. Recent evidences suggest that some glycoproteins may require sorting receptors for efficient exit from the endoplasmic reticulum. These receptors need the presence of calcium or other metal ions for their native activity. The three-dimensional structure of such a receptor, p58/ERGIC-53, has been recently solved by x-ray crystallography, which is a mannose-selective lectin and contains two Ca(2+) ions. Homology search in the sequence databases indicates a large number of proteins which bear varying degrees of homology in a wide spectrum of species with this receptor. In this study we have systematically searched for such genes which are potential candidates for acting as mannose mediated glycoprotein receptors in various species as initially inferred from their amino acid sequence homology. Structures of a number of proteins have been predicted using knowledge-based homology modeling, and their ability to act as the glycoprotein receptor has been explored by examining the nature of sugar binding site. Tetramer of mannose was docked in the binding pockets of the modeled structures followed by energy minimization and molecular dynamics to obtain most probable structures of the complexes. Properties of these modeled complexes were studied to examine the nature of physicochemical forces involved in the complex formation and compared with p58/ERGIC-53-mannose complex. PMID- 16691508 TI - Search for fucose binding domains in recently sequenced hypothetical proteins using molecular modeling techniques and structural analysis. AB - The crystal structure of a fucose-binding lectin from the bacteria Pseudomonas aeruginosa in complex with alpha-L-fucose has been recently determined. It is a tetramer; each monomer displays a nine-stranded, antiparallel, beta-sandwiched arrangement and contains two calcium ions that mediate the binding of fucose in a recognition mode unique among protein-carbohydrate interactions. In search of this type of unique interactions in other newly discovered protein sequences, we have used molecular modeling techniques to predict and analyze the 3-D structures of some proteins, which exhibited reasonable degree of homology with the amino acid sequence of the bacterial protein. A BLAST search with the sequence of Pseudomonas aeruginosa as query in the non-redundant sequence database identified four proteins from different species, three organisms from bacteria and one from archaea. We have modeled the structures of these proteins as well as those of the complexes with carbohydrates and studied the nature of physicochemical forces involved in the complex formation both in presence and absence of calcium. The calcium-binding loops have been found to be highly conserved both in terms of primary and tertiary structures in these proteins, although a less acidic character is observed in Photorhabdus lectin due to the absence of two aspartic acid residues on the calcium-binding loop which also resulted in lower binding affinity. All these structures exhibited highly negative electrostatic environment in the vicinity of the calcium-binding loops which was essential for neutralizing the positive charges of two closely situated Ca(+2) ions. The comparison of the binding affinities of some monosaccharides other than fucose, e.g. mannose and fructose, showed higher binding energies confirming the fucose specificity of these proteins. PMID- 16691509 TI - Transcriptional activation of tyrosinase gene by human placental sphingolipid. AB - The sphingolipids, a class of complex bioactive lipids, are involved in diverse cellular functions such as proliferation, differentiation, and apoptosis as well as growth inhibition. Recently sphingosylphosphorylcholine (SPC), sphingosine-1 phosphate (S1P), and C2-ceramide (C2-Cer), sphingolipid containing acetic acid are emerging as melanogenic regulators. A bioactive sphingolipid (PSL) was isolated from hydroalcoholic extract of fresh term human placenta and it induced melanogenesis in an in vitro culture of mouse melanoma B16F10 cells. Tyrosinase, the rate-limiting enzyme for melanogenesis, is required to be upregulated for the increased melanin production. The expression of tyrosinase, both at protein as well as mRNA level, was higher in the PSL treated B16F10 cells as evidenced by Western blot and RT-PCR analysis. Actinomycin D and cycloheximide, inhibitors of transcription and translation, respectively, inhibited PSL-induced tyrosinase activity and its protein expression showing decrease in melanogenesis, correspondingly. The activity of GFP coupled tyrosinase promoter was upregulated in transfected B16F10 cells after treating with PSL as determined by fluorescence microscopy, fluorometric analysis, and Western blot. These results, thus, suggested that PSL upregulated tyrosinase gene expression at transcription level through promoter activation to show increased melanogenesis. Therefore, PSL as an inducer of melanogenesis might account for the recovery of pigment in depigmentation disorder. PMID- 16691511 TI - Environmental correlates of physiological variables in marsupials. AB - We analyzed body temperature (T(b)), basal metabolic rate (BMR), wet thermal conductance (C(wet)), and evaporative water loss (EWL) of marsupials by conventional and phylogenetically corrected regression. Allometric effects were substantial for BMR, C(wet), and EWL but not T(b). There was a strong phylogenetic signal for mass and all physiological traits. A significant phylogenetic signal remained for BMR, C(wet), and EWL even after accounting for the highly significant phylogenetic signal of mass. T(b), BMR, C(wet), and EWL allometric residuals were correlated with some diet, distribution, and climatic variables before and after correction for phylogeny. T(b) residuals were higher for marsupials from arid environments (high T(a) and more variable rainfall). The fossorial marsupial mole had a lower-than-expected T(b) residual. The allometric slope for BMR was 0.72-0.75. Residuals were consistently related to distribution aridity and rainfall variability, with species from arid and variable rainfall habitats having a low BMR, presumably to conserve energy in a low-productivity environment. The nectarivorous honey possum had a higher-than-expected BMR. For C(wet), the allometric slope was 0.55-0.62; residuals were related to diet, with folivores having low and insectivores high C(wet) residuals. The allometric slope for EWL was 0.68-0.73. EWL residuals were consistently correlated with rainfall variability, presumably facilitating maintenance of water balance during dry periods. PMID- 16691512 TI - Extreme plasticity in thermoregulatory behaviors of free-ranging black-tailed prairie dogs. AB - In the natural environment, hibernating sciurids generally remain dormant during winter and enter numerous deep torpor bouts from the time of first immergence in fall until emergence in spring. In contrast, black-tailed prairie dogs (Cynomys ludovicianus) remain active throughout winter but periodically enter short and shallow bouts of torpor. While investigating body temperature (T(b)) patterns of black-tailed prairie dogs from six separate colonies in northern Colorado, we observed one population that displayed torpor patterns resembling those commonly seen in hibernators. Five individuals in this population experienced multiple torpor bouts in immediate succession that increased in length and depth as winter progressed, whereas 16 prairie dogs in five neighboring colonies remained euthermic for the majority of winter and entered shallow bouts of torpor infrequently. Our results suggest that these differences in torpor patterns did not result from differences in the physiological indicators that we measured because the prairie dogs monitored had similar body masses and concentrations of stored lipids across seasons. Likewise, our results did not support the idea that differences in overwinter T(b) patterns between prairie dogs in colonies with differing torpor patterns resulted from genetic differences between populations; genetic analyses of prairie dog colonies revealed high genetic similarity between the populations and implied that individuals regularly disperse between colonies. Local environmental conditions probably played a role in the unusual T(b) patterns experienced by prairie dogs in the colony where hibernation-like patterns were observed; this population received significantly less rainfall than neighboring colonies during the summer growing seasons before, during, and after the year of the winter in which they hibernated. Our study provides a rare example of extreme plasticity in thermoregulatory behaviors of free-ranging prairie dogs and provides evidence contrary to models that propose a clear delineation between homeothermy, facultative torpor, and hibernation. PMID- 16691513 TI - The development of endothermy during pouch life in the eastern barred bandicoot (Perameles gunnii), a marsupial. AB - Marsupials are born ectothermic and gradually become endothermic during pouch occupancy. In order to study the timing of this transition, we measured the metabolic rates of eight pouch-young eastern barred bandicoots (Perameles gunnii) at 25 degrees C (the thermoneutral zone of adults), at 35 degrees C (pouch temperature), and after injection with norepinephrine. From 5 to 7 wk of age, oxygen consumption (VO(2)) was higher at 35 degrees C than at 25 degrees C. At 8 wk, VO(2) was significantly higher at 25 degrees C than at 35 degrees C. Norepinephrine administration at 5 and 6 wk had no effect on metabolic rate, but at 8 wk, near pouch vacation, it resulted in a significant increase in metabolic rate. Our results indicate that signs of thermoregulation for P. gunnii begin at 7 wk, and at 8 wk the species is endothermic, that is, able to increase heat production in response to both cold and norepinephrine. The appearance of fur, first seen at the age of 6 wk and fully developed at 8 wk, coincides with the onset of endothermy. PMID- 16691514 TI - Mitochondrial metabolism in hibernation: metabolic suppression, temperature effects, and substrate preferences. AB - We compared liver and skeletal muscle mitochondrial function among activity states to characterize regulated reversible metabolic suppression in the mammalian hibernator Spermophilus tridecemlineatus. At 37 degrees C, succinate oxidation was 70% lower in the liver mitochondria from torpid animals than in those from summer-active animals or in animals arousing from torpor. Respiration was very sensitive to temperature (Q(10) 5.8-9.8), and when measured at 25 degrees or 5 degrees C there was no difference among the three states. Liver mitochondria from summer-active animals oxidized pyruvate and beta hydroxybutyrate at higher rates than those from torpid animals, and flux through complex 4 of the electron transport chain was about three- and fivefold higher than flux through complexes 2-4 and complexes 1-4, respectively. In the hibernating and arousing animals there was no difference in flux through complexes 2-4 and complex 4, suggesting a downregulation of cytochrome c oxidase in liver mitochondria during the hibernation season. Muscle mitochondrial respiration did not differ between the torpid and summer-active states in any of the parameters measured. The data support a regulated, reversible decrease of liver (but not muscle) mitochondrial oxidative phosphorylation in hibernating ground squirrels. PMID- 16691515 TI - Physiology and behavior influence lactation efficiency in northern elephant seals (Mirounga angustirostris). AB - The efficiency with which mothers convert acquired energy into milk is a key determinant of the magnitude of parental investment in mammals; however, the mechanisms underlying lactation efficiency are poorly understood. Investigations on northern elephant seals have shown lactation efficiency, measured as the proportion of total energy expenditure that goes to the pup as milk, increases with age. In a cross-sectional study the physiological and behavioral determinants of lactation efficiency were investigated in eight young and seven prime (older) elephant seals by conducting behavioral observations and collecting milk, blood, and tissue on days 3 and 22 of lactation. Milk composition, circulating fatty acid and triglyceride concentrations, and mammary and blubber lipoprotein lipase activity were determined. Prime females had significantly greater percent milk fat and circulating fatty acids on day 3 than did young females, but these differences disappeared by day 22. The ability for prime females to produce higher-energy milk early in lactation may allow them to increase lactation efficiency by increasing the rate of energy transfer. In addition, prime females spent significantly more time resting. A combination of reduced activity and more rapid energy delivery likely explains the increase in lactation efficiency with age found in a previous study. PMID- 16691516 TI - Ruminal fermentation and fill change with season in an arctic grazer: responses to hyperphagia and hypophagia in muskoxen (Ovibos moschatus). AB - We studied castrated adult muskoxen fed a standard diet of grass hay and supplement throughout the year to determine seasonal changes in digesta passage, fill, and fermentation without the confounding effects of reproductive demands or changes in food quality. Although food intake increased by 74% between spring and autumn, mean retention times of fluid and particulate digesta markers were maintained between seasons in both the rumen (9-13 h) and the intestines (27-37 h). The rumen contained 84.5% of digesta and accounted for 79% of dry matter digestion in the whole digestive tract. Ruminal fluid space and whole-gut digesta fill increased by 31%-34%, while ruminal rates of in situ degradation increased by more than 100% between spring and autumn for cellulose and hemicellulose. Hyperphagia in autumn was accompanied by increased bacterial counts in ruminal fluid (30%), declines in ruminal pH, and increases in the concentration of fermentation acids (16%) when compared with spring hypophagia. Consumption of fresh hay and supplement increased the concentrations of acids most markedly during winter and spring when bacterial counts were low. Low food intakes in winter and spring may limit the microbial population, whereas hyperphagia in autumn may foster a much more active microflora that requires consistent supplies of substrate. Plasticity of fill and fermentation in muskoxen minimizes winter costs and maximizes nutrients and energy gained from coarse forages in small home ranges throughout the year. PMID- 16691517 TI - Body mass and clutch size may modulate prolactin and corticosterone levels in eiders. AB - Altered body condition, increased incubation costs, and egg loss are important proximate factors modulating bird parental behavior, since they inform the adult about its remaining chances of survival or about the expected current reproductive success. Hormonal changes should reflect internal or external stimuli, since corticosterone levels (inducing nest abandonment) are known to increase while body condition deteriorates, and prolactin levels (stimulating incubation) decrease following egg predation. However, in a capital incubator that based its investment on available body reserves and naturally lost about half of its body mass during incubation, corticosterone should be maintained at a low threshold to avoid protein mobilization for energy supply. This study focused on the regulation of corticosterone and prolactin release in such birds during incubation, when facing egg manipulation (control, reduced, or increased) or a stressful event. Blood samples were taken before and after clutch manipulation and at hatching. Corticosterone levels were determined before and after 30 min of captivity. Female eiders exhibited a high hypothalamic-pituitary-adrenal sensitivity, plasma concentration of corticosterone being increased by four- to fivefold following 30 min of captivity. The adrenocortical response was not modified by body mass loss but was higher in birds for which clutch size was increased. In the same way, females did not show different prolactin levels among the experimental groups. However, when incubation started, prolactin levels were correlated to body mass, suggesting that nest attendance is programmed in relation to the female initial body condition. Moreover, due to an artifactual impact of bird manipulation, increased baseline corticosterone was associated with a prolactin decrease in the control group. These data suggest that, in eiders, body mass and clutch size modification can modulate prolactin and corticosterone levels, which cross-regulate each other in order to finely control incubation behavior. PMID- 16691518 TI - Scaling of Na+,K+-ATPase molecular activity and membrane fatty acid composition in mammalian and avian hearts. AB - We have examined Na(+),K(+)-ATPase molecular activity and membrane fatty acid composition in the heart of six mammalian and eight avian species ranging in size from 30 g in mice to 280 kg in cattle and 13 g in zebra finches to 35 kg in emus, respectively. Na(+),K(+)-ATPase activity scaled negatively with body mass in both mammals and birds. In small mammals, the elevated enzyme activity was related to allometric changes in both the concentration and molecular activity (turnover rate) of Na(+),K(+)-ATPase enzymes, while in small birds, higher Na(+),K(+) ATPase activity appeared to result primarily from an increased molecular activity of individual enzymes. The unsaturation index of cardiac phospholipids scaled negatively with body mass in both groups, while a significant allometric increase in monounsaturate content was observed in the larger mammals and birds. In particular, the relative content of the highly polyunsaturated docosahexaenoic acid (22:6n-3) displayed the greatest variation, scaling negatively with body mass and varying greater than 40-fold in both mammals and birds. Membrane fatty acid profile was correlated with Na(+),K(+)-ATPase molecular activity in both mammals and birds, suggesting a potential association between membrane lipid composition and the activity of membrane-bound enzymes in the hearts of endotherms. PMID- 16691519 TI - Metabolic routing of dietary nutrients in birds: effects of diet quality and macronutrient composition revealed using stable isotopes. AB - During fall migration many songbirds switch from consuming primarily insects to consuming mostly fruit. Fruits with more carbohydrates and less protein may be sufficient to rebuild expended fat stores, but such fruits may be inadequate to replace catabolized protein. We manipulated the concentrations and isotopic signatures of macronutrients in diets fed to birds to study the effects of diet quality on metabolic routing of dietary nutrients. We estimated that approximately 45% and 75%, respectively, of the carbon in proteinaceous tissue of birds switched to high- or low-protein diets came from nonprotein dietary sources. In contrast, we estimated that approximately 100% and 20%-80%, respectively, of the nitrogen in proteinaceous tissues of birds switched to high- or low-protein diets was attributable to dietary protein. Thus, the routing and assimilation of dietary carbon and nitrogen differed depending on diet composition. As a result, delta (15)N of tissues collected from wild animals that consume high-quality diets may reliably indicate the dietary protein source, whereas delta (13)C of these same tissues is likely the product of metabolic routing of carbon from several macronutrients. These results have implications for how isotopic discrimination is best estimated and how we can study macronutrient routing in wild animals. PMID- 16691520 TI - Yolk testosterone stimulates growth and immunity in house finch chicks. AB - Female birds deposit variable amounts of androgens, such as testosterone, into the yolks of their eggs. Evidence suggests that yolk androgens play an important role in the determination of offspring phenotype. While androgens are generally regarded as anabolic and immunosuppressive, studies of the behavioral and physiological effects of yolk androgens on offspring of several avian species have been conflicting, leaving the adaptive significance associated with deposition patterns of yolk androgens unclear. We injected either a physiological dose of testosterone or a control vehicle into house finch (Carpodacus mexicanus) eggs and examined the effects of these injections on offspring growth and immunity. Two days after hatching, nestlings from eggs treated with testosterone were significantly larger than nestlings from eggs treated with a control injection, suggesting a stimulatory effect of yolk androgens in early development. By 8 d after hatching, however, this effect disappeared, and chicks from the two treatment groups were similar in size. Nestlings in the testosterone treatment group showed a significantly larger swelling response to phytohemagglutinin than control nestlings 15 d after hatching, which is close to fledging. Overall, our observations show that when food resources are abundant, testosterone stimulates both early growth and immunity in developing house finches. PMID- 16691521 TI - Capture stress and the bactericidal competence of blood and plasma in five species of tropical birds. AB - In wild birds, relatively little is known about intra- or interspecific variation in immunological capabilities, and even less is known about the effects of stress on immune function. Immunological assays adaptable to field settings and suitable for a wide variety of taxa will prove most useful for addressing these issues. We describe a novel application of an in vitro technique that measures the intrinsic bacteria-killing abilities of blood. We assessed the capacities of whole blood and plasma from free-living individuals of five tropical bird species to kill a nonpathogenic strain of E. coli before and after the birds experienced an acute stress. Killing invasive bacteria is a fundamental immune function, and the bacteria-killing assay measures constitutive, innate immunity integrated across circulating cell and protein components. Killing ability varied significantly across species, with common ground doves exhibiting the lowest levels and blue crowned motmots exhibiting the highest levels. Across species, plasma killed bacteria as effectively as whole blood, and higher concentrations of plasma killed significantly better. One hour of acute stress reduced killing ability by up to 40%. This assay is expected to be useful in evolutionary and ecological studies dealing with physiological and immunological differences in birds. PMID- 16691522 TI - Hematological health state indices predict local survival in a small passerine bird, the great tit (Parus major). AB - In birds, it has been shown that reproductive effort may impair parental condition, while the relation of different condition indices to subsequent survival is still poorly understood. In this study, we measured body mass and various hematological condition indices in breeding great tits in relation to local survival. Number and quality of nestlings and the occurrence of second broods, potentially reflecting parents' breeding effort, were also considered in analyses. The great tits, both male and female, that returned the following year had had a higher albumin/globulin ratio, lower plasma globulin concentration, and a lower heterophile/lymphocyte ratio during breeding in the preceding year, compared to those who did not return. Surviving males (but not females) also had had a higher level of circulating lymphocytes, compared to nonsurvivors. There was no correlation between breeding effort and survival. We conclude that better immunological state and lower stress in great tits during breeding were positively related to their survival probability. PMID- 16691523 TI - Energetics of lizard embryos are not canalized by thermal acclimation. AB - In some species of ectotherms, temperature has little or no effect on the amount of energy expended during embryonic development. This phenomenon can result from either of two mechanisms: (1) a shorter incubation period at higher temperatures, which offsets the expected increase in metabolic rate, or (2) a compensatory decrease in the rate at which embryos expend energy for maintenance. To distinguish the relative importance of these two mechanisms, we quantified the acute and chronic effects of temperature on embryonic metabolism in the eastern fence lizard (Sceloporus undulatus). First, we measured metabolic rates of individual embryos at 27 degrees, 31 degrees, and 34 degrees C. Second, we examined the capacity for thermal acclimation by measuring the metabolic rates of embryos at 30 degrees C, after a period of incubation at either 28 degrees or 32 degrees C. As with adult reptiles, the metabolic rates of embryos increased with an acute increase in temperature; the Q(10) of metabolic rate from 27 degrees to 34 degrees C was 2.1 (+/-0.2). No evidence of thermal acclimation was observed either early or late in development. In S. undulatus, a shorter incubation period at higher temperatures appears to play the primary role in canalizing the energy budget of an embryo, but a reduction in the cost of growth could play a secondary role. PMID- 16691524 TI - Evolution of viviparity in sceloporine lizards: in utero PO2 as a developmental constraint during egg retention. AB - Reptilian viviparity evolves through selection for increasingly prolonged egg retention within the oviduct. In the majority of sceloporine lizard species, however, egg retention past the normal time of oviposition results in retarded or arrested embryonic development. In this study, we tested the hypothesis that the amount of embryonic development normally attained in utero is directly related to in utero oxygen partial pressure (PO(2)). The three species of sceloporine lizards we used are characterized by developmental arrest (Urosaurus ornatus), retarded development (Sceloporus virgatus), and normal development (Sceloporus scalaris) when eggs are retained. We incubated eggs of these species for 10 d under conditions that simulated retention in the oviduct at a range of experimental oxygen partial pressures (PO(2)). We estimated in utero PO(2) from a standard curve generated from the stage and dry mass of experimental embryos incubated for 10 d at known PO(2). The standard curve was then used to predict the PO(2) associated with the observed rate of development of embryos retained in utero. The results of this study showed that the degree of embryonic development attained in utero during egg retention was positively associated with in utero PO(2). The results indicate that oxygen availability in utero is associated with interspecific differences in the capacity to support intrauterine development in sceloporine lizards. PMID- 16691525 TI - Metabolic response to feeding in Tupinambis merianae: circadian rhythm and a possible respiratory constraint. AB - The diurnal tegu lizard Tupinambis merianae exhibits a marked circadian variation in metabolism that is characterized by the significant increase in metabolism during part of the day. These increases in metabolic rate, found in the fasting animal, are absent during the first 2 d after meal ingestion but reappear subsequently, and the daily increase in metabolic rate is added to the increase in metabolic rate caused by digestion. During the first 2 d after feeding, priority is given to digestion, while on the third and following days, the metabolic demands are clearly added to each other. This response seems to be a regulated response of the animal, which becomes less active after food ingestion, rather than an inability of the respiratory system to support simultaneous demands at the beginning of digestion. The body cavity of Tupinambis is divided into two compartments by a posthepatic septum (PHS). Animals that had their PHS surgically removed showed no significant alteration in the postprandial metabolic response compared to tegus with intact PHS. The maximal metabolic increment during digestion, the relative cost of meal digestion, and the duration of the process were virtually unaffected by the removal of the PHS. PMID- 16691526 TI - The dogfish shark (Squalus acanthias) increases both hepatic and extrahepatic ornithine urea cycle enzyme activities for nitrogen conservation after feeding. AB - Urea not only is utilized as a major osmolyte in marine elasmobranchs but also constitutes their main nitrogenous waste. This study investigated the effect of feeding, and thus elevated nitrogen intake, on nitrogen metabolism in the Pacific spiny dogfish Squalus acanthias. We determined the activities of ornithine urea cycle (O-UC) and related enzymes in liver and nonhepatic tissues. Carbamoyl phosphate synthetase III (the rate-limiting enzyme of the O-UC) activity in muscle is high compared with liver, and the activities in both tissues increased after feeding. The contribution of muscle to urea synthesis in the dogfish body appears to be much larger than that of liver when body mass is considered. Furthermore, enhanced activities of the O-UC and related enzymes (glutamine synthetase, ornithine transcarbamoylase, arginase) were seen after feeding in both liver and muscle and were accompanied by delayed increases in plasma urea, trimethylamine oxide, total free amino acids, alanine, and chloride concentrations, as well as in total osmolality. The O-UC and related enzymes also occurred in the intestine but showed little change after feeding. Feeding did not change the rate of urea excretion, indicating strong N retention after feeding. Ammonia excretion, which constituted only a small percentage of total N excretion, was raised in fed fish, while plasma ammonia did not change, suggesting that excess ammonia in plasma is quickly ushered into synthesis of urea or protein. In conclusion, we suggest that N conservation is a high priority in this elasmobranch and that feeding promotes ureogenesis and growth. Furthermore, exogenous nitrogen from food is converted into urea not only by the liver but also by the muscle and to a small extent by the intestine. PMID- 16691527 TI - Regulation of the cardiorespiratory system of common carp (Cyprinus carpio) during severe hypoxia at three seasonal acclimation temperatures. AB - Little is known of the cardiorespiratory control mechanisms utilized by hypoxia tolerant teleost fish to tolerate prolonged periods (h) of near anoxic exposure. Here, we report on the cardiorespiratory control mechanisms of the common carp Cyprinus carpio L. during normoxia and prolonged, severe hypoxic (<0.3 mg O(2) L( 1)) exposure at acclimation temperatures of 5 degrees C, 10 degrees C, and 15 degrees C. Through serial intra-arterial injections of alpha - and beta adrenergic, cholinergic, and purinergic antagonists while measuring cardiac output (Q), heart rate (f(H)), ventral aortic blood pressure, and respiration rate, we established that autonomic cardiovascular and respiratory control was preserved during severe hypoxia at all three acclimation temperatures and contributed to downregulation of cardiorespiratory activity. Specifically, inhibitory cholinergic tone mediated up to 76% reductions in f(H) and Q during hypoxia, whereas the accompanying arterial hypotension was attenuated by an upregulation of an alpha -adrenergically mediated peripheral vasoconstriction. Despite the overall cardiac downregulation, a large, stimulatory cardiac beta adrenergic tone was present during prolonged, severe hypoxia, possibly to protect the heart from attendant acidotic conditions. Purinergic blockade, following alpha -adrenergic and cholinergic antagonists, showed that the hypoxic ventilatory depression, which reversed the 2.3- to 7.7-fold increases in respiration rate that occurred with the onset of hypoxia, was a result of purinergic inhibition at all three acclimation temperatures. In contrast, purinergic inhibition of cardiac activity during hypoxia might be important only at 5 degrees C. Finally, given that cardiac power output was reduced 72%-87% during prolonged, severe hypoxia and that glycolysis yields approximately 94% less ATP per mole glucose than oxidative phosphorylation, it seems unlikely that the common carp sufficiently reduces its cardiac energy demand to a level to preclude activation of a partial Pasteur effect. This means that glycogen stores will be used and waste products will accumulate at faster rates, a finding that may help explain why the common carp cannot tolerate such extended periods of severe hypoxia (weeks to months) at cold acclimation temperatures as the freshwater turtle, which is able to reduce its cardiac energy demand to a level that does not require a Pasteur effect and also blunts autonomic cardiovascular control. PMID- 16691528 TI - Body protein stores and isotopic indicators of N balance in female reindeer (Rangifer tarandus) during winter. AB - We studied bred and unbred female reindeer (Rangifer tarandus tarandus) during 12 wk of winter when ambient temperatures were low and nitrogen (N) demand for fetal growth is highest in pregnant females. Animals were fed a complete pelleted diet ad lib. that contained 2.54% N in dry matter that was 80% +/- 2% (X +/- SD) digestible. Female reindeer lost 64% +/- 14% of body fat but gained 34% +/- 11% of lean mass from 10 wk prepartum to parturition. These changes were equivalent to average balances of -14.14 +/- 2.35 MJ d(-1) and 10 +/- 3 g N d(-1). Blood cells, serum, and urine declined in (15)N/(14)N in late winter as body protein was gained from the diet. Blood cells of newborn calves were more enriched in (15)N and (13)C than that of their mothers, indicating the deposition of fetal protein from maternal stores. To quantify pathways of N flow in reindeer, N balance was measured by confining animals to cages for 10 d at 4 wk from parturition. N balance was inversely related to (15)N/(14)N in urea-N but not related to (15)N/(14)N of blood cells, creatinine, and feces. The proportion of urea-N derived from body protein increased above 0.46 as N balance fell below 200 mg N kg(-0.75) d(-1). Proportions of urea-N from body protein were -0.01 +/- 0.21 in pregnant females before and after caging and were consistent with average body protein gain in winter. Storage of protein allows reindeer and caribou to tolerate diets that are low in N without impairing fetal development. PMID- 16691529 TI - Secretion of water and ions by malpighian tubules of larval mosquitoes: effects of diuretic factors, second messengers, and salinity. AB - The effects of changes in the salinity of the rearing medium on Malpighian tubule fluid secretion and ion transport were examined in larvae of the freshwater mosquito Aedes aegypti and the saltwater species Ochlerotatus taeniorhynchus. For unstimulated tubules of both species, the K(+) concentration of secreted fluid was significantly lower when larvae were reared in 30% or 100% seawater (O. taeniorhynchus only), relative to tubules from freshwater-reared larvae. The Na(+) concentration of secreted fluid from unstimulated tubules of O. taeniorhynchus reared in 30% or 100% seawater was higher relative to tubules from freshwater-reared larvae. The results suggest that changes in salinity of the larval rearing medium lead to sustained changes in ion transport mechanisms in unstimulated tubules. Furthermore, alterations of K(+) transport may be utilized to either conserve Na(+) under freshwater (Na(+)-deprived) conditions or eliminate more Na(+) in saline (Na(+)-rich) conditions. The secretagogues cyclic AMP [cAMP], cyclic GMP [cGMP], leucokinin-VIII, and thapsigargin stimulated fluid secretion by tubules of both species. Cyclic AMP increased K(+) concentration and decreased Na(+) concentration in the fluid secreted by tubules isolated from O. taeniorhynchus larvae reared in 100% seawater. Interactions between rearing salinity and cGMP actions were similar to those for cAMP. Leucokinin-VIII and thapsigargin had no effect on secreted fluid Na(+) or K(+) concentrations. Results indicate that changes in rearing medium salinity affect the nature and extent of stimulation of fluid and ion secretion by secretagogues. PMID- 16691530 TI - Light induces an increase in the pH of and a decrease in the ammonia concentration in the extrapallial fluid of the giant clam Tridacna squamosa. AB - The objective of this study was to examine whether 12 h of light exposure would lead to an increase in the pH of and a decrease in the concentration of total ammonia in the extrapallial fluid of the giant clam Tridacna squamosa. We also aimed to elucidate indirectly whether movements of ammonia and/or protons (H(+)) occurred between the extrapallial fluid and the outer mantle epithelium. The pH of the extrapallial fluid of T. squamosa exposed to 12 h of light was significantly higher than that of clams exposed to 12 h of darkness. Conversely, the total ammonia concentration in the extrapallial fluid of the former was significantly lower than that of the latter. In addition, the glutamine content in the mantle adjacent to the extrapallial fluid of clams exposed to 12 h of light was significantly greater than that of clams exposed to 12 h of darkness. These results suggest that in the extrapallial fluid of T. squamosa exposed to light, NH(3) combined with H(+) as NH(+)(4) and that NH(+)(4) was transported into the mantle and used as a substrate for glutamine formation. Injection of NH(4)Cl into the extrapallial fluid led to an instantaneous increase in the total ammonia concentration therein, but the total ammonia concentration decreased subsequently and returned to the control value within 1 h. This is in support of the proposition that NH(+)(4) could be transported from the extrapallial fluid to the mantle. Injection of HCl into the extrapallial fluid led to an instantaneous decrease in the pH of the extrapallial fluid. However, there was a significant increase in pH within 1 h in light or darkness, achieving a partial recovery toward the control pH value. The increase in pH within this 1-h period in light or darkness was accompanied by a significant decrease in the total ammonia concentration in the extrapallial fluid, which supports the proposition that H(+) could be transported in combination with NH(3) as NH(+)(4). Therefore, our results prompt a reexamination of the previous proposition that the removal of H(+) by NH(3) can facilitate calcification in molluscs in general and an investigation of the relationship between H(+) removal through NH(+)(4) transport and light-enhanced calcification in T. squamosa. PMID- 16691531 TI - Allometry, antilog transformations, and the perils of prediction on the original scale. AB - Biologists often use allometric equations that take the form of power functions (e.g., Y = aM(b), where M stands for mass and a and b are empirically fitted constants). Typically, these allometric equations are fitted by taking the antilog of log-log regressions. Predictions from these allometric equations are biased, and the bias my be appreciable. Methods for making predictions that correct for the bias are available, but they have rarely, if ever, been used by ecological and evolutionary physiologists. Just as physiologists would not use an instrument that was not properly calibrated, they should not use allometric equations to make predictions unless they account for the bias of those predictions. We analyzed 20 interspecific and 10 intraspecific data sets. We compared predictions from standard allometric equations with those from several alternative methods. Our analyses suggest that the bias of predictions from interspecific data sets may be substantial. For the intraspecific data sets we analyzed, the bias was likely to be small. Biologists, including ecological and evolutionary physiologists, should exercise care when using allometric equations to make predictions, particularly given that methods to adjust for bias are easily implemented. PMID- 16691532 TI - Macrophage populations and cardiac sympathetic denervation during L-NAME-induced hypertension in rats. AB - The rat model of hypertension induced by prolonged treatment with Nomega-nitro-L arginine methyl ester (L-NAME) has been extensively used. However, the effects on cardiac autonomic innervation are unknown. Here, the cardiac sympathetic innervation is analyzed in parallel with myocardial lesions and leukocyte infiltration during L-NAME (40 mg/Kg body weight/day, orally) treatment. The occurrence of cardiomyocyte hypertrophy, a controversial matter, is also addressed. Degenerating cardiomyocytes and focal inflammation occurred one day after treatment. Inflammatory lesions became gradually more frequent until day 7. At day 14 fibroblast-like cells were outstanding. Interstitial and perivascular connective tissue increased from day 28 on. In the left ventricle, cardiomyocyte hypertrophy occurred only around the damaged area during the first 14 days. After 28 days, it became more widespread. In the right ventricle, the hypertrophic cardiomyocytes were restricted to damaged areas. Significant reduction of the noradrenergic nerve terminals occurred from day 3 to 28. The area occupied by ED1+ (hematogenous) macrophages increased until day 7, and dropped to control levels by day 10. ED2+ (resident) macrophages increased from day 3 to 7 and remained higher than control values up to day 77. Animals receiving both L- NAME and aminoguanidine (AG), an inducible nitric oxide synthase (iNOS) inhibitor (65 mg/Kg body weight/day, orally), showed significant decrease in the nitrite serum levels, sympathetic denervation and macrophage infiltration at day 7. No denervation was detectable at day 14 of double treatment, using subcutaneous AG. Our findings favor a role for ED1+ macrophages and iNOS in the hypertension induced denervation process. PMID- 16691533 TI - Megakaryocytic features useful for the diagnosis of myeloproliferative disorders can be obtained by a novel unsupervised software analysis. AB - An unsupervised method for megakaryocyte detection and analysis is proposed, in order to validate supplementary tools which can be of help in supporting the pathologist in the classification of Philadelphia negative chronic myeloproliferative disorders with thrombocytosis. The experiment was conducted on high power magnification photomicrographs taken from hematoxylin-and-eosin 3 micrometer thick sections of formalin fixed, paraffin embedded bone marrow biopsies from patients with reactive thrombocytosis or chronic myeloproliferative disorders. Each megakaryocyte has been isolated in the photos through an image segmentation process, mainly based on mathematical morphology and wavelet analysis. A set of features (e.g. area, perimeter and fractal dimension of the cell and its nucleus, shape complexity via elliptic Fourier transform, and so on) is used to characterize the disorders and discriminate between essential thrombocythemia and idiopathic myelofibrosis. Features related to the general contour of the cell like cytoplasmic area and perimeter are good markers in distinguishing between normal or reactive and pathologic megakaryocytes while nuclear features and global circularity are helpful in the differential diagnosis between ET and prefibrotic IMF. The method proposed should be considered as a fast preprocessing tool for the diagnostic phase and its use can be extended to solve different object recognition problems. PMID- 16691534 TI - Increased collagen deposition correlated with lung destruction in human emphysema. AB - BACKGROUND: To study the relationship between collagen amount and degree of emphysema as assessed by mean linear intercept (Lm) and correlating these with lung function test workup in patients with and without COPD. METHODS: Lung function tests were assessed in 16 smokers or ex-smokers and 1 non-smoker in order to separate them into two groups: COPD (FEV1/FVC lower than 70%) and non COPD. A piece of lung tissue was used to analyse the collagen amount (HYP) by means of a colorimetric method. Morphometry was assessed to divide patients into two groups according to Lm: Lm > 260 micrometers was considered non-emphysema and Lm < 260 mm mild-emphysema. RESULTS: The non-emphysema group had a mean Lm value of 246.08+/-3.12 micrometers and the mild-emphysema group of 276.29+/-4.26 micrometers. The amount of hydroxyproline was significantly higher in the mild emphysema group than in the non-emphysema group (7.82+/-0.67 vs. 5.50+/-0.54 microgram/g tissue). There was a clear positive correlation between Lm and HYP (r=0.55) and a negative correlation between Lm and DlCO (R=-0.5092). No correlation was found between the functional test and HYP, nor were there significant differences between COPD and non-COPD patients for Lm and HYP. CONCLUSIONS: Emphysema is associated with collagen deposition in the lungs, and air space size correlates with the amount of lung collagen even when there is no emphysema. PMID- 16691535 TI - Histological changes and micronucleus induction in the zebra mussel Dreissena polymorpha after paraquat exposure. AB - The herbicide paraquat (PQ), still widely used in developing countries, represents a serious risk factor for human and environmental health. To test the sublethal effects of PQ on the freshwater bivalve Dreissena polymorpha, mussels were exposed to 0.125, 0.250, 0.500 mg/L for 7 and 14 days and histologically screened. PQ's genotoxic potential was also determined in haemocytes by the micronucleus, MN, assay. At concentrations > or = 0.250 mg/L, severe lesions, such as cellular vacuolation, lysis and thinness of the germinative epithelia were observed in the digestive gland and testis. A positive trend between the number of granulocytes and all PQ concentrations was observed in both gonads and digestive glands, addressing the inflammatory capacity of this herbicide on these tissues. Mussels exposed to PQ also exhibited a significant MN induction. The spontaneous MN frequencies ranged from 2.75 to 0.425 per cent, while PQ-induced MN rates in treated mussels were between 3.50 and 1.250 per cent. The histopathological effects on the digestive and reproductive systems, as well as the MN induction in the haemocytes, confirmed the cytotoxic and genotoxic effects of PQ also in D. polymorpha. PMID- 16691536 TI - Abnormal expression and clinicopathologic significance of p120-catenin in lung cancer. AB - The aim of this study was to investigate the relationship between the expression of p120ctn in human lung squamous cell carcinoma, adenocarcinoma and its clinicopathologic significance. The expression of p120ctn in tumors and adjacent normal lung tissues from 143 patients was examined by immunohistochemistry and Western blot. Expression of p120ctn occurs mainly in the cell membrane of normal bronchial mucosa. Abnormal expression of p120ctn, including cytoplasmic and reduced membranous expression, was found in 114 of 143 specimens (79.7%) and was significantly associated with poor differentiation, high TNM stage, and lymph node metastasis (P<0.05 for each) but not with histologic subtype. The Kaplan Meier survival test revealed that abnormal expression of p120ctn was related to poor survival (P<0.001). A Cox regression analysis revealed that abnormal p120ctn expression was an independent factor in predicting patient survival (P=0.024). Compared with that in normal lung tissues, membranous protein level was lower in tumors (P=0.003). Abnormal expression of p120ctn is associated with tumor progression and poor prognosis in lung squamous cell carcinoma and adenocarcinoma. Reduced expression or even the absence of p120ctn isoform 1 and 3 in tumor cell membranes may be responsible for the abnormal expression of p120ctn that has been found in lung cancer. PMID- 16691537 TI - MUC1 cytoplasmic tail detection using CT33 polyclonal and CT2 monoclonal antibodies in breast and colorectal tissue. AB - The immunohistochemical detection (IHC) of MUC1-CT employing a polyclonal antibody (CT33) in relation to CT2 monoclonal antibody (MAb) was analyzed. Western blot (WB) was used to determine the molecular mass of CT. MATERIALS AND METHODS: We studied 163 breast and 89 colorectal cancer specimens, 10 breast and 14 colorectal benign conditions, and 12 breast and 20 colorectal normal samples. From each tumor sample, subcellular fractions were obtained and analyzed by SDS PAGE and WB. A nonparametric statistical analysis was employed; data were standardized and a Kendall-Tau correlation was applied. RESULTS: By IHC, 146/163 (90%) and 151/163 (93%) of breast cancer were positive with CT33 and CT2, respectively; a statistically significant correlation was obtained (t=0.5199). Seven out of ten (70%) benign breast specimens were positive with CT33 while all samples stained with CT2; in normal breast sample tissues, all were positive with both Abs. In colorectal cancer samples, both antibodies stained 47/89 (53%) samples; CT2 reacted in 13/14 (93%) of benign samples while CT33 showed a positive reaction in 9/14 (64%) of benign specimens. In normal samples, CT2 showed staining in 17/20 (85%) of samples and CT33 was reactive in 12/20 (60%). By WB, in breast and colorectal cancer samples, similar results were obtained with both antibodies: a main band at about 30kDa which represents the smaller subunit. CONCLUSION: CT33 polyclonal antibody has demonstrated its efficacy to detect MUC1 in breast and colorectal cancer tissues with similar reactivity to CT2. It is worthwhile to affirm that CT33 is a good indicator of MUC1 expression. PMID- 16691538 TI - Study of microvessel density and the expression of the angiogenic factors VEGF, bFGF and the receptors Flt-1 and FLK-1 in benign, premalignant and malignant prostate tissues. AB - PURPOSE: Vascular endothelial growth factor (VEGF) is an angiogenic factor that stimulates endothelial cell growth and enhances vascular permeability. VEGF exerts its action by binding to the specific cell surface receptors, fms-like tyrosine kinase 1 (Flt-1) and fetal liver kinase 1 (FLK/KDR). In tumor angiogenesis, Vascular endothelial growth factor stimulates endothelial cells to produce Basic fibroblastic growth factor (bFGF), which further enhances angiogenic activity. Very little information on the expression of VEGF, bFGF, and the receptors Flt-1 and FLK/KDR is available. Herein, we evaluate the expression of these angiogenic factors and receptors in normal prostate, high grade prostate intraepithelial neoplasia (HGPIN) and prostatic cancer (CaP). MATERIALS AND METHODS: 58 selected surgical specimens exhibiting areas of normal prostate, HGPIN, and CaP were evaluated for microvessel density, and for VEGF, bFGF, Flt-1 and FLK/KDR protein expression by immunohistochemistry. Results were correlated with pathological data. RESULTS: There was a statistically significant increase in the microvessel density and in the expression of the angiogenic factors VEGF, bFGF and the receptors FLK/KDR and Flt-1, in the premalignant and malignant tissues in comparison with normal prostatic glands. Microvessel density also correlated with higher Gleason grade, pathological stage and the expression of the receptors FLK/KDR and Flt-1. CONCLUSIONS: The "initiation switch" of angiogenesis was observed to be an early event consistent with the recruitment of new vasculature into high grade PIN lesions and it increased in the progression of prostatic cancer. PMID- 16691539 TI - Recent advances in the treatment of colon cancer. AB - Colorectal cancer is one of the leading causes of cancer-related deaths worldwide. Although surgical resection is still the only treatment capable of curing colon cancer, adjuvant therapy continues to play an important role in preventing recurrence and metastasis. In recent years remarkable progress has been made in the treatment of colon cancer. This review discusses recent advances in adjuvant therapy for colon cancer, including chemotherapy, immunotherapy, antiangiogenic therapy and apoptosis induction. In the meantime, molecular therapy is also elucidated in the above methods. All these new advances will provide new promises for patients of colon cancer. PMID- 16691540 TI - Mad2 and spindle assembly checkpoint function during meiosis I in mammalian oocytes. AB - During mammalian mitosis, a proofreading network called the spindle assembly checkpoint (SAC) is indispensable for ensuring the fidelity of chromosome segregation. An inhibitory SAC signal is deputed to inhibits mitotic cell-cycle progression in response to misaligned chromosomes until such imperfections are rectified thereby ensuring equitable chromosome partitioning to daughter cells. Amongst the cast of SAC proteins, mitotic arrest deficient 2 (Mad2) plays a leading role in transducing the SAC signal. The aneuploidy and cancer predispositions of individuals who harbour genetic mutations in SAC genes emphasise the in vivo significance of this surveillance mechanism. In humans, congenital aneuploidies such as Down's syndrome demonstrate an exponential increase with advancing female age. Although largely the result of female meiosis I errors, the molecular entities that succumb with age in oocytes remain elusive. Declining oocyte SAC function could plausibly contribute to such errors. Until recently however, convincing evidence for a functional SAC in mammalian oocytes during meiosis I was unforthcoming. Here I review the evidence regarding the SAC in female mammalian meiosis I and how our understanding of this system has evolved in recent years. This review will focus on Mad2 as this is the SAC protein that has been most comprehensively investigated. PMID- 16691541 TI - In vitro maturation of human primordial ovarian follicles: clinical significance, progress in mammals, and methods for growth evaluation. AB - As cancer treatment improves, more young women of reproductive age are surviving, but they suffer from infertility as a consequence of the radiation and chemotherapy. Human ovarian tissue containing immature primordial follicles has been successfully cryopreserved. The ultimate aim of this technique is to induce ovarian function by re-plantation of ovarian tissue or, further into the future, by in vitro maturation (IVM) of the oocytes derived from the cryopreserved-thawed ovarian tissue, followed by routine in vitro fertilization. IVM of primordial follicles from young cancer survivors would avoid the risk of cancer re transmission by the ovarian grafts. The present review discusses the current achievements in IVM of female germ cells and primordial ovarian follicles and the attempts to improve their development by adding various factors to the culture medium. The established methods for the evaluation of survival and growth in culture are also discussed: follicular counts, immunocytochemical methods, transmission electron microscopy, fluorescent viability markers and endocrine assays. Although the development of IVM systems is still in its infancy, researchers need to pursue their approach step-by-step, especially with regard to factors that might be involved in the activation of the ovarian follicles or female germ cells. The final measure of success will be the ability of the in vitro matured oocytes to fertilize and produce healthy offsprings. The availability of such treatment will probably lead to its demand not only by cancer patients but by other women as well. PMID- 16691542 TI - The sorting and trafficking of lysosomal proteins. AB - For a long time lysosomes were considered terminal organelles involved in the degradation of different substrates. However, this view is rapidly changing by evidence demonstrating that these organelles and their content display specialized functions in addition to the degradation of substances. Many lysosomal proteins have been implicated in specialized cellular functions and disorders such as antigen processing, targeting of surfactant proteins, and most lysosomal storage disorders. To date, about fifty lysosomal hydrolases have been identified, and the majority of them are targeted to the lysosomes via the mannose-6-phosphate receptor (M6P-Rc). However, recent studies on the intracellular trafficking of the non-enzymic lysosomal proteins prosaposin and GM2 activator (GM2AP) demonstrated that they use an alternative receptor termed "sortilin". Existing evidence suggests that some hydrolases traffic to the lysosomes in a mannose 6-phophate-indepentend manner. The possibility that sortilin is implicated in the targeting of some soluble hydrolases, as well as the consequences of this process, is addressed in the present review. PMID- 16691543 TI - Sorption-desorption of flucarbazone and propoxycarbazone and their benzenesulfonamide and triazolinone metabolites in two soils. AB - Sorption-desorption interactions of pesticides with soil determine the availability of pesticides in soil for transport, plant uptake and microbial degradation. These interactions are affected by the physical and chemical properties of the pesticide and soil and, for some pesticides, their residence time in the soil. While sorption-desorption of many herbicides has been characterised, very little work in this area has been done on herbicide metabolites. The objective of this study was to characterise sorption-desorption of two sulfonylaminocarbonyltriazolinone herbicides, flucarbazone and propoxycarbazone, and their benzenesulfonamide and triazolinone metabolites in two soils with different physical and chemical properties. K(f) values for all four chemicals were greater in clay loam soil, which had higher organic carbon and clay contents than loamy sand. K(f-oc) ranged from 29 to 119 for the herbicides and from 42 to 84 for the metabolites. Desorption was hysteretic in every case. Lower desorption in the more sorptive system might indicate that hysteresis can be attributed to irreversible binding of the molecules to soil surfaces. These data show the importance of characterisation of both sorption and desorption of herbicide residues in soil, particularly in the case of prediction of herbicide residue transport. In this case, potential transport of sulfonylaminocarbonyltriazolinone herbicide metabolites would be overpredicted if parent chemical soil sorption values were used to predict transport. PMID- 16691544 TI - Trypsin in colorectal cancer: molecular biological mechanisms of proliferation, invasion, and metastasis. AB - Trypsin is involved in colorectal carcinogenesis and promotes proliferation, invasion, and metastasis. Although a well-known pancreatic digestive enzyme, trypsin has also been found in other tissues and various cancers, most importantly of the colorectum. Moreover, colorectal cancers with trypsin expression have a poor prognosis and shorter disease-free survival. Biological understanding of how trypsin causes cancer progression is emerging. It seems to act both directly and indirectly through a 'proteinase-antiproteinase-system', and by activation of other proteinase cascades. Invasion of the basal membrane by cancer cells may be promoted directly by trypsin digestion of type I collagen. Trypsin activates, and is co-expressed with matrix metalloproteinases (MMPs), which are known to facilitate invasion and metastasis. MMP-2, MMP-7, and MMP-9 are co-expressed together with trypsin and seem to be of particular importance in proliferation, progression, and invasion. MMPs may play a role in both conversion from adenoma to carcinoma, and in the initiation of invasion and metastasis. Co segregation of trypsin and MMPs within the tumour environment is important for the activation of MMPs, and may explain the deleterious effect of trypsin on prognosis in colorectal cancer. Trypsin and proteinase-activated receptor 2 (PAR 2) act together in an autocrine loop that promotes proliferation, invasion, and metastasis through various mechanisms, of which prostaglandin synthesis is important. Stimulated by trypsin, both MMP and PAR-2 may activate the mitogenic MAPK-ERK pathway through activation of the epidermal growth factor receptor. Experimental trypsin inhibition is feasible but not very effective, and trypsin as a target for clinical therapy is unlikely to be successful owing to its universal distribution. However, as the pathways of trypsin and co-activated protein cascades emerge, biological understanding of colorectal carcinogenesis will be further illuminated and may pave the way for prognosticators, predictors, and novel targets of therapy. PMID- 16691545 TI - Potential of imidacloprid to control four species of psocids (Psocoptera: Liposcelididae) infesting stored grain. AB - The effectiveness of the neonicotinoid insecticide imidacloprid was evaluated against four psocid pests of stored grain. This research was undertaken because of the growing importance of psocids in stored grain and the need to identify methods for their control. The mortality and reproduction of adults of Liposcelis bostrychophila Badonnel, L. entomophila (Enderlein), L. decolor (Pearman) and L. paeta Pearman in wheat treated with imidacloprid were determined. There were five application rates (0.5, 1, 2, 5 and 10 mg AI kg(-1) grain) and an untreated control. There were significant effects of application rate on both adult mortality and reproduction for all four species, but the effect of imidacloprid was sometimes more pronounced on reproduction. Imidacloprid was most effective against L. bostrychophila, with 100% adult mortality after 7 d at 5 mg AI kg(-1), 14 d at 2 mg AI kg(-1) and 28 d at 0.5 and 1 mg AI kg(-1). No live progeny were produced at 2 mg AI kg(-1). For L. decolor, there was 100% adult mortality after 28 d at 10 mg AI kg(-1) and no live progeny were produced at 2 mg AI kg(-1). For L. entomophila, there was 100% adult mortality after 14 d at 10 mg AI kg(-1) and 28 d at 2 and 5 mg AI kg(-1). No live progeny were produced at 10 mg AI kg(-1). At 10 mg AI kg(-1) there was 100% mortality of L. paeta adults after 28 d exposure and no live progeny developed. Because reproduction at some application rates occurred only in the first 14 d of exposure, it is concluded that the application rate leading to population extinction was 1 mg AI kg(-1) for L. bostrychophila, 2 mg AI kg(-1) for L. decolor and L. entomophila and 5 mg AI kg( 1) for L. paeta. This study shows that imidacloprid has potential as a grain protectant to control all four Liposcelis species in stored grain. PMID- 16691546 TI - Effect of various extracts and a polysaccharide from the edible mycelia of Cordyceps sinensis on cellular and humoral immune response against ovalbumin in mice. AB - In this study, the edible mycelia of Cordyceps sinensis (Berk.) Sacc. were sequentially extracted by petroleum ether, ethyl acetate, ethanol and ultrasonic water-bath (75 degrees C), and the water extract was further isolated by Sephadex G-100 to afford a petroleum ether extract (PE), ethyl acetate extract (EAE), ethanol extract (EE), glycoprotein (GP) and a purified polysaccharide (PS). In combination with component analysis, the isolated PS showed d-Glc, d-Man, l-Ara and d-Gal in a molar ratio of 8:90:1:1. The average molecular weight of PS was determined as approximately 8.3 x 10(4). The immunomodulatory potentials of these samples (PE, EAE, EE, GP and PS) at three dose levels on the cellular and humoral immune responses of ICR mice against ovalbumin (OVA) were studied. ICR mice were immunized subcutaneously with 100 microg OVA alone or with 100 microg OVA dissolved in saline containing these samples (100, 200 or 400 microg) on days 1 and 15. Two weeks later (day 28), concanavalin A (Con A)- and OVA-stimulated splenocyte proliferation and OVA-specific antibody in serum were investigated. These samples, except for PS, significantly enhanced the Con A- and OVA-induced splenocyte proliferation in OVA-immunized mice at a suitable dose (p < 0.05 or p < 0.01). OVA-specific IgG, IgG1 and IgG2b antibody levels in serum were significantly enhanced by these extracts and PS compared with the OVA control group (p < 0.05, p < 0.01 or p < 0.001). PMID- 16691547 TI - Inhibitory effect of polar oregano extracts on aldose reductase and soybean lipoxygenase in vitro. AB - The effect of methanol and aqueous methanol extract of Origanum vulgare L. ssp. hirtum on aldose reductase and soybean lipoxygenase was investigated. The results revealed a promising potential of oregano for preventing diabetes complications in the long term and an antiinflammatory efficacy by inhibiting soybean lipoxygenase. PMID- 16691548 TI - Kinetic analysis and modelling of the allosteric behaviour of liver and muscle glycogen phosphorylases. AB - Allosteric enzymes have very complex kinetic behaviours which are primarily interpreted through simplified models. To describe the functional properties of liver and muscle glycogen phosphorylase isozymes we have developed an experimental strategy based on the measurements of initial reaction rates in the presence of different concentrations of the effectors glucose-1-phosphate and methyl-xanthines. Using the extensive structural information available for the two glycogen phosphorylase conformers T (inactive) and R (active) with different ligands, we have applied the Monod-Wyman-Changeux model and analysed the results in the context of the exclusive binding of the inhibitors to the T state, meanwhile the substrate glucose-1-phosphate binds to both, the R and T states. The kinetic analysis shows a good agreement between our model and the results obtained from the glycogen phosphorylases and inhibitors included in this study, which demonstrates the validity of the approach described here. PMID- 16691549 TI - Identification of new Golgi complex specific proteins by direct organelle proteomic analysis. AB - The Golgi complex is in the crossroad of the endocytic and secretory pathways. Its function is to post-translationally modify and sort proteins and lipids, and regulate the membrane balance in the cell. To understand the structure-function relationship of the Golgi complex the Golgi proteome has to be identified first. We have used a direct organelle proteomic analysis to identify new Golgi complex proteins. Enriched stacked Golgi membrane fractions from rat livers were isolated, and the proteins from these membranes were subsequently digested into peptides. The peptides were fractionated by cation-exchange chromatography followed by protein identification by automated capillary-LC/ESI-MS/MS analysis and database searches. Two different search programs, ProID and MASCOT were used. This resulted in a total of 1125 protein identifications in two experiments. In addition to the known Golgi resident proteins, a significant number of unknown proteins were identified. Some of these were further characterized in silico using different programs to provide insight into their structure, intracellular localization and biological functions. The Golgi localization of two of these newly identified proteins was also confirmed by indirect immunofluorescence. PMID- 16691550 TI - Proteomic profiling of fibroblasts reveals a modulating effect of extracellular calumenin on the organization of the actin cytoskeleton. AB - CREC proteins constitute a family of EF-hand calcium binding proteins localized to the secretory pathway. Calumenin is the only member known to be secreted. Recently, it was shown that thrombin-activated thrombocytes liberate calumenin, which also is found in atherosclerotic lesions but not in normal vasculature. To study the possible effects of calumenin extracellularly, we used proteomic profiling of fibroblasts cultured in absence and in presence of calumenin. Using 2-DE and MS/MS, we show that normal fibroblasts contain several 28-29-kDa N terminal and a 16-kDa C-terminal fragment of beta- or gamma-actin. Extracellularly added calumenin decreases the levels of both the N-terminal and C terminal actin fragments, and, in addition, decreases the expression level of septin 2, which interacts with the actin cytoskeleton and is involved in cytokinesis. Labeling of S-phase fibroblasts with bromo-2'deoxy-uridine indicates that calumenin added to the medium also modulates the cell cycle. Our study thus indicates that calumenin may have an autocrine or a paracrine effect on the cells in its vicinity, and, therefore, may be involved in the pathophysiology of thrombosis or in wound healing. PMID- 16691551 TI - Determination of the phosphorylation level and deamidation susceptibility of equine beta-casein. AB - beta-Casein was isolated from Haflinger mare's milk by RP-HPLC, and displayed microheterogeneity by urea-electrophoresis and 2-DE probably due to a variable degree of phosphorylation. To investigate the degree of phosphorylation, the primary structure of equine beta-casein was determined by tryptic hydrolysis and MS of peptides released and by MS of the protein treated by alkaline phosphatase. The molecular mass found for the apo-form of Haflinger mare's beta-casein (25 514 +/- 3 Da) was close to the theoretical mass of the reported sequence (GenBank AAG43954) modified by insertion of a region (residues 27-34) encoded by an exon sometimes out-spliced (25 511.40 Da). Hence, the beta-casein isolated from Haflinger mare's milk corresponded to a variant of 226 amino acid residues. The latter was composed by highly multi-phosphorylated isoforms with three to seven phosphate groups, and pIs, determined by 2-DE, ranging from 4.74 to 5.30. Moreover, the equine beta-casein was able to deamidate spontaneously, at the level of Asn in the potential deamidation motif (135)Asn-Gly(136). Approximately 80% of the protein was deamidated after 96 h of incubation under physiological conditions. PMID- 16691552 TI - Impact of the accessory gene regulatory system (Agr) on extracellular proteins, codY expression and amino acid metabolism in Staphylococcus epidermidis. AB - The quorum-sensing system Agr is part of a complex regulatory network of gene expression in staphylococci. This study presents the effect of an agr mutation on a biofilm-forming Staphylococcus epidermidis isolate by employing proteome and transcriptome analysis. The agr mutant exhibited a significantly lowered amount of extracellular proteins: amongst others SspA, AtlE, GehD and the phenol soluble modulins PSM1/2. Cytoplasmic proteome analysis and expression profiling indicated that the agr inactivation led to a strongly altered regulation of metabolism and virulence. Most strikingly, expression of CodY, a global regulator of virulence and stationary phase gene expression, was decreased in the agr mutant. In this respect, homologous genes known to be controlled by CodY in Bacillus subtilis and Lactococcus lactis were found to be up-regulated in the S. epidermidis agr mutant. The combined data show that wild-type and agr mutant differ with respect to amino acid biosynthesis and oligopeptide transport, carbohydrate utilization, as well as GMP and IMP interconversion. Due to the varying physiological properties S. epidermidis agr mutants, which often occur spontaneously, might be capable of colonizing alternative ecological niches in the human host and could, therefore, have an advantage in adapting to changing environmental conditions. PMID- 16691553 TI - Proteomic analysis of the eukaryotic parasite Encephalitozoon cuniculi (microsporidia): a reference map for proteins expressed in late sporogonial stages. AB - The microsporidian Encephalitozoon cuniculi is a unicellular obligate intracellular parasite considered as an emerging opportunistic human pathogen. The differentiation phase of its life cycle leads to the formation of stress resistant spores. The E. cuniculi genome (2.9 Mbp) having been sequenced, we undertook a descriptive proteomic study of a spore-rich cell population isolated from culture supernatants. A combination of 2-DE and 2-DE-free techniques was applied to whole-cell protein extracts. Protein identification was performed using an automated MALDI-TOF-MS platform and a nanoLC-MS/MS instrument. A reference 2-DE map of about 350 major spots with multiple isoforms was obtained, and for the first time in microsporidia, a large set of unique proteins (177) including proteins with unknown function in a proportion of 25.6% was identified. The data are mainly discussed with reference to secretion and spore structural features, energy and carbohydrate metabolism, cell cycle control and parasite survival in the environment. PMID- 16691554 TI - Multivariate analysis of protein profiles of metal hyperaccumulator Thlaspi caerulescens accessions. AB - Thlaspi caerulescens is increasingly acknowledged as one of the best models for studying metal hyperaccumulation in plants. In order to study the mechanisms underlying metal hyperaccumulation, we used proteomic profiling to identify differences in protein intensities among three T. caerulescens accessions with pronounced differences in tolerance, uptake and root to shoot translocation of Zn and Cd. Proteins were separated using two-dimensional electrophoresis and stained with SYPRO Orange. Intensity values and quality scores were obtained for each spot by using PDQuest software. Principal component analysis was used to test the separation of the protein profiles of the three plant accessions at various metal exposures, and to detect groups of proteins responsible for the differences. Spot sets representing individual proteins were analysed with the analysis of variance and non-parametric Kruskal-Wallis test. Clearest differences were seen among the Thlaspi accessions, while the effects of metal exposures were less pronounced. The 48 tentatively identified spots represent core metabolic functions (e.g. photosynthesis, nitrogen assimilation, carbohydrate metabolism) as well as putative signalling and regulatory functions. The possible roles of some of the proteins in heavy metal accumulation and tolerance are discussed. PMID- 16691555 TI - Proteomic characterization of acid stress response in Synechocystis sp. PCC 6803. AB - A comparative proteomic analysis using 2-DE coupled with MALDI-MS and LC-MS/MS was performed in Synechocystis sp. PCC 6803 to identify protein candidates involved in acid stress response in cyanobacteria. Comparison of soluble proteins from the cytoplasmic fraction of cells grown on media set at pH 7.5 and 5.5 using 2-DE identified four proteins, which showed significant changes in the abundance. Surprisingly, several general stress proteins, either the heat shock family proteins or chaperonins, did not show perceptible fold changes in response to acidity. Compared to the cytoplasmic proteome, the periplasmic proteome showed remarkable changes as a function of external pH. Protein expression profiling at different external pH, i.e., 9.0, 7.5, 6.0 and 5.5, allowed classifying the periplasmic proteins depending on their preferential expression patterns towards acidity or alkalinity. Among the acid- and base-induced proteins, oxalate decarboxylase and carbonic anhydrase were already known for their role in pH homeostasis. Several unknown proteins from the periplasm, that showed significant changes in response to pH, provide ideal targets for further studies in understanding pH stress response in cyanobacteria. This study also identified 14 novel proteins, hitherto unknown from the periplasmic space of Synechocystis. PMID- 16691556 TI - DuraPrep and the risk of fire during tracheostomy. AB - BACKGROUND: DuraPrep is a widely used, alcohol-based surgical prep solution. The risk of surgical fire associated with incomplete drying of this agent in the context of electrosurgical procedures has been described previously. To date, there have been no reports of fire during tracheostomy associated with a flammable prep agent before entering the airway. We describe an operating room fire occurring during awake tracheostomy associated with the use of DuraPrep. METHODS: A 62-year-old man with copious body hair underwent tracheostomy in the operating room. The neck was prepared with DuraPrep surgical solution, and after at least 3 minutes, the operative field was draped. Activation of electrocautery ignited a fire, and the patient was burned on his neck and shoulders. RESULTS: The fire was extinguished, and the patient recovered from both the tracheostomy and the burns. CONCLUSION: This case illustrates that DuraPrep should be avoided in the hirsute patient, because body hair interferes with drying of this solution and increases the risk of fire. PMID- 16691557 TI - Importance of molecular analysis in detecting cervical lymph node metastasis in head and neck squamous cell carcinoma. AB - BACKGROUND: Because of the impact of nodal status on treatment and survival in squamous cell carcinoma of the head and neck, accurate staging of cervical lymph nodes is critical. This article explores the value of molecular analyses in the detection of cervical lymph node metastasis. METHODS: A review of the literature was carried out and combined with our own experience regarding the role of molecular analyses in detecting cervical lymph node metastasis. RESULTS: Few studies have demonstrated the diagnostic and prognostic relevance of molecular analysis in detecting tumor cells in lymph nodes. Nodal staging was improved by the use of molecular techniques; when compared with histopathologic examination, however, the small sample size of these studies did not allow definitive conclusions. CONCLUSIONS: Molecular analysis is exquisitely sensitive in detecting very small cancer deposits within lymph nodes. It provides an oncologic basis that may be used to guide therapy and influence outcomes. It should be recommended for diagnostic use in controlled studies of patients without evidence of lymph node metastasis on routine hematoxylin-eosin-stained sections. The clinical significance of these types of metastases, however, must be determined with carefully designed and controlled prospective clinical trials. PMID- 16691558 TI - Association of the A870G cyclin D1 gene polymorphism with genetic susceptibility to nasopharyngeal carcinoma. AB - BACKGROUND: Nasopharyngeal cancer (NPC) is multifactorial, and the genetic background may be a crucial etiologic factor. Cyclin D1 (CCND1) is a key regulator of the cell cycle, and its altered activity is associated with the development of cancer. METHODS: We analyzed the A870G CCND1 polymorphism by polymerase chain reaction/restriction fragment length polymorphism (PCR-RFLP) in 281 individuals, including 94 patients with NPC and 187 healthy individuals. RESULTS: Our results indicate that individuals carrying two G alleles have a 2.17 fold increase in the risk for the development of NPC (odds ratio [OR], 2.17; 95% confidence interval [CI], 1.19-3.98; p = .016). Age-adjusted logistic regression analysis confirmed this association (adjusted odds ratio [aOR], 2.14; 95% CI, 1.14-4.04; p = .018). Multivariate analysis demonstrates an independent association between GG CCND1 genotype (aOR, 2.06), male sex (aOR, 2.66), and age at diagnosis (aOR, 2.02) regarding the development of undifferentiated NPC. The proportion of NPC cases attributable to the GG CCND1 genotype was 14.76%. CONCLUSIONS: Our results may be important in the definition of a biologic predictive profile for the development of NPC within our population. PMID- 16691559 TI - Reconstruction of the columella using the prefabricated reverse flow submental flap: A case report. AB - BACKGROUND: The reconstruction of columellar defects is still a challenging procedure because of limited local and regional flap options and the characteristics of the anatomy of this site. Although a number of methods are available to repair nasal columella defects, no treatment of choice ensuring an excellent texture- and color-matched tissue in one stage has been determined to date. METHOD: In this case, we used a reverse-flow submental island flap prefabricated with the costal cartilage for the reconstruction of a complex columellar defect. RESULT: The flap survived completely with reversible venous congestion. The cosmetic result and nasal respiratory function were acceptable during the follow-up time of 6 months. CONCLUSION: We propose that the prefabricated reverse submental flap may be an alternative among the surgical options for columellar defects. This flap may also be considered in the reconstructive repertoire of other composite defects of the head and neck region. PMID- 16691560 TI - Altered immune response to CNS viral infection in mice with a conditional knock down of macrophage-lineage cells. AB - Neuroadapted Sindbis Virus (NSV) is a neuronotropic virus that causes hindlimb paralysis in susceptible mice and rats. The authors and others have demonstrated that though death of infected motor neurons occurs, bystander death of uninfected neurons also occurs and both contribute to the paralysis that ensues following infection. The authors have previously shown that the treatment of NSV-infected mice with minocycline, an inhibitor that has many functions within the central nervous system (CNS), including inhibiting microglial activation, protects mice from paralysis and death. The authors, therefore, proposed that microglial activation may contribute to bystander death of motor neurons following NSV infection. Here, the authors tested the hypothesis using a conditional knock-out of activated macrophage-lineage cells, including endogenous CNS macrophage cells. Surprisingly, ablation of these cells resulted in more rapid death and similar weakness in the hind limbs of NSV-infected animals compared with that of control animals. Several key chemokines including IL-12 and monocyte chemoattractant protein-1 (MCP-1) did not become elevated in these animals, resulting in decreased infiltration of T lymphocytes into the CNS of the knock-down animals. Either because of the decreased macrophage activation directly or because of the reduced immune cell influx, viral replication persisted longer within the nervous system in knock-down mice than in wild type mice. The authors, therefore, conclude that although macrophage-lineage cells in the CNS may contribute to neurodegeneration in certain situations, they also serve a protective role, such as control of viral replication. PMID- 16691561 TI - Mouse coat color mutations: from fancy mice to functional genomics. AB - Mouse coat color mutations have a long history in biomedical research. The viable and visible phenotype of most coat color mutations has made the pigment cell, the melanocyte, an ideal system for genetic, molecular, and cellular analysis. Molecular cloning and analysis of many of the different coat color mutations have revealed the roles of a diverse range of genes, and today we know more about the pathways and proteins that regulate the development and function of pigment cells than we know about most other cell types in mammalian organisms. Coat color mutations have also provided novel insights into stem cell biology and human diseases, including melanoma. In the future, it will be important to build on this history and knowledge by taking advantage of the extensive repertoire of recently developed genome-wide methodologies, available genomic information, and the powerful methods that have been developed for modifying the mouse genome to systematically dissect the development and function of this important cell type. The usefulness of coat color mutations has just begun to emerge. PMID- 16691562 TI - Bmp2 and Gata4 function additively to rescue heart tube development in the absence of retinoids. AB - We used the vitamin A-deficient (VAD) quail model to investigate the retinoid dependent mechanism that regulates heart tube development. We showed previously that decreased levels of Gata4 in cardiogenic mesoderm and endoderm correlate with the cardiomyopathy caused by VAD, but that this could be rescued by transplanting normal anterior endoderm. Bmp2 is a known cardiogenic factor that is expressed normally in lateral plate mesoderm and cardiac-associated pharyngeal endoderm. Here we show that (like Gata4) transcripts encoding Bmp2 and BMP dependent signaling activity are decreased throughout the heart-forming region of the VAD embryo. Addition of Bmp2 protein or forced expression of Gata4 in cultured VAD embryos leads to a partial rescue of the cardiomyopathy, and addition of both Bmp2 and Gata4 has an additive positive effect. Our data are consistent with a requirement for retinoid signaling to maintain expression of Bmp2, which regulates Gata4, and in addition acts with Gata4 to regulate genes important for normal morphogenesis of the primitive heart tube. PMID- 16691563 TI - Modeling H3 histone N-terminal tail and linker DNA interactions. AB - Molecular dynamics computer simulations were performed for the 25-residue N terminal tail of the H3 histone protein in the proximity of a DNA segment of 10 base pairs (bp), representing a model for the linker DNA in chromatin. Several least biased configurations were used as initial configurations. The secondary structure content of the protein was increased by the presence of DNA close to it, but the locations of the secondary motifs were different for different initial orientations of the DNA grooves with respect to the protein. As a common feature to all simulations, the electrostatic attraction between negatively charged DNA and positively charged protein was screened by the water solvent and counterbalanced by the intrinsic compaction of the protein due to hydrophobic effects. The protein secondary structure limited the covering of DNA by the protein to 4-5 bp. The degree of compaction and charge density of the bound protein suggests a possible role of H3 tail in a nonspecific bending and plasticity of the linker DNA when the protein is located in the crowded dense chromatin. PMID- 16691564 TI - Foxg1 is required for morphogenesis and histogenesis of the mammalian inner ear. AB - The forkhead genes are involved in patterning, morphogenesis, cell fate determination, and proliferation. Several Fox genes (Foxi1, Foxg1) are expressed in the developing otocyst of both zebrafish and mammals. We show that Foxg1 is expressed in most cell types of the inner ear of the adult mouse and that Foxg1 mutants have both morphological and histological defects in the inner ear. These mice have a shortened cochlea with multiple rows of hair cells and supporting cells. Additionally, they demonstrate striking abnormalities in cochlear and vestibular innervation, including loss of all crista neurons and numerous fibers that overshoot the organ of Corti. Closer examination shows that some anterior crista fibers exist in late embryos. Tracing these fibers shows that they do not project to the brain but, instead, to the cochlea. Finally, these mice completely lack a horizontal crista, although a horizontal canal forms but comes off the anterior ampulla. Anterior and posterior cristae, ampullae, and canals are reduced to varying degrees, particularly in combination with Fgf10 heterozygosity. Compounding Fgf10 heterozygotic effects suggest an additive effect of Fgf10 on Foxg1, possibly mediated through bone morphogenetic protein regulation. We show that sensory epithelia formation and canal development are linked in the anterior and posterior canal systems. Much of the Foxg1 phenotype can be explained by the participation of the protein binding domain in the delta/notch/hes signaling pathway. Additional Foxg1 effects may be mediated by the forkhead DNA binding domain. PMID- 16691565 TI - Proteolytic cleavage of versican during cardiac cushion morphogenesis. AB - The proteoglycan versican is essential to the formation of endocardial cushion mesenchyme by epithelial-mesenchymal transformation (EMT). A potentially important factor in the regulation of versican activity during cushion EMT is proteolysis by ADAMTS metalloproteinases. Using antibodies to the DPEAAE neoepitope created by ADAMTS proteolysis of versican, we detected the amino terminal 70-kDa versican cleavage fragment in cardiac cushions. Initially (i.e., 9.5 days post coitum [dpc]), the fragment is associated with endocardial cells undergoing EMT and with newly derived mesenchymal cells. ADAMTS-1 and its cofactor fibulin-1 were also associated with these cells. As cushions become increasingly populated with mesenchymal cells (10.5-12.5 dpc), the fragment remains asymmetrically distributed compared with the pattern of total versican. Highest levels of the fragment are present in regions immediately subjacent to the endocardium characterized as having densely packed, rounded cells, lacking cellular protrusions. With further development (i.e., 12.5-14.5 dpc), the pattern of fragment distribution within cushions broadens to include the ECM surrounding loosely packed mesenchymal cells in the cushion core. Together, the findings reveal that versican proteolysis leading to the production of the 70-kDa fragment is integral to the formation and differentiation of endocardial cushion mesenchyme. PMID- 16691566 TI - Alpha N-catenin deficiency causes defects in axon migration and nuclear organization in restricted regions of the mouse brain. AB - Alpha N-catenin is a cadherin-binding protein, widely expressed in the nervous system; and it plays a crucial role in cadherin-mediated cell-cell adhesion. Here we report the effects of alpha N-catenin gene deficiency on brain morphogenesis. In addition to the previously reported phenotypes, we found that some of the axon tracts did not normally develop, in particular, axons of the anterior commissure failed to cross the midline, migrating, rather, to ectopic places. In restricted nuclei, a population of neurons was missing or their laminar arrangement was distorted. The ventricular structures were also deformed. These results indicate that alpha N-catenin has diverse roles in the organization of the central nervous system, but only in limited portions of the brain. PMID- 16691567 TI - Ratio control variate method for efficiently determining high-dimensional model representations. AB - The High-Dimensional Model Representation (HDMR) technique is a family of approaches to efficiently interpolate high-dimensional functions. RS(Random Sampling)-HDMR is a practical form of HDMR based on randomly sampling the overall function, and utilizing orthonormal polynomial expansions to approximate the RS HDMR component functions. The determination of the expansion coefficients for the component functions employs Monte Carlo integration, which controls the accuracy of the RS-HDMR interpolation. The control variate method is an established approach to improve the accuracy of Monte Carlo integration. However, this method is often not practical for an arbitrary function f(x) because there is no general way to find the analytical control variate function h(x), which needs to be very similar to f(x). In this article, we show that truncated RS-HDMR expansions can be used as h(x) for arbitrary f(x), and a more stable iterative ratio control variate method was developed for the determination of the expansion coefficients for the RS-HDMR component functions. As the asymptotic error (standard deviation) of the estimator given by the ratio control variate method is proportional to 1/N(sample size), it is more efficient than the direct Monte Carlo integration, whose error is proportional to 1/square root(N). In an illustration of a four dimensional atmospheric model a few hundred random samples are sufficient to construct an RS-HDMR expansion by the ratio control variate method with an accuracy comparable to that obtained by direct Monte Carlo integration with thousands of samples. PMID- 16691568 TI - RM1: a reparameterization of AM1 for H, C, N, O, P, S, F, Cl, Br, and I. AB - Twenty years ago, the landmark AM1 was introduced, and has since had an increasingly wide following among chemists due to its consistently good results and time-tested reliability--being presently available in countless computational quantum chemistry programs. However, semiempirical molecular orbital models still are of limited accuracy and need to be improved if the full potential of new linear scaling techniques, such as MOZYME and LocalSCF, is to be realized. Accordingly, in this article we present RM1 (Recife Model 1): a reparameterization of AM1. As before, the properties used in the parameterization procedure were: heats of formation, dipole moments, ionization potentials and geometric variables (bond lengths and angles). Considering that the vast majority of molecules of importance to life can be assembled by using only six elements: C, H, N, O, P, and S, and that by adding the halogens we can now build most molecules of importance to pharmaceutical research, our training set consisted of 1736 molecules, representative of organic and biochemistry, containing C, H, N, O, P, S, F, Cl, Br, and I atoms. Unlike AM1, and similar to PM3, all RM1 parameters have been optimized. For enthalpies of formation, dipole moments, ionization potentials, and interatomic distances, the average errors in RM1, for the 1736 molecules, are less than those for AM1, PM3, and PM5. Indeed, the average errors in kcal x mol(-1) of the enthalpies of formation for AM1, PM3, and PM5 are 11.15, 7.98, and 6.03, whereas for RM1 this value is 5.77. The errors, in Debye, of the dipole moments for AM1, PM3, PM5, and RM1 are, respectively, 0.37, 0.38, 0.50, and 0.34. Likewise, the respective errors for the ionization potentials, in eV, are 0.60, 0.55, 0.48, and 0.45, and the respective errors, in angstroms, for the interatomic distances are 0.036, 0.029, 0.037, and 0.027. The RM1 average error in bond angles of 6.82 degrees is only slightly higher than the AM1 figure of 5.88 degrees, and both are much smaller than the PM3 and PM5 figures of 6.98 degrees and 9.83 degrees, respectively. Moreover, a known error in PM3 nitrogen charges is corrected in RM1. Therefore, RM1 represents an improvement over AM1 and its similar successor PM3, and is probably very competitive with PM5, which is a somewhat different model, and not fully disclosed. RM1 possesses the same analytical construct and the same number of parameters for each atom as AM1, and, therefore, can be easily implemented in any software that already has AM1, not requiring any change in any line of code, with the sole exception of the values of the parameters themselves. PMID- 16691569 TI - The Jahn-Teller and pseudo-Jahn-Teller effects in the anion photoelectron spectroscopy of B3 cluster. AB - The photodetachment spectroscopy of B3- anion is theoretically studied with the aid of a quantum dynamical approach. The theoretical results are compared with the available experimental photoelectron spectra of B3-. Both B3- and B3 possess D(3h) symmetry at the equilibrium configuration of their electronic ground state. Distortion of B3 along its degenerate vibrational mode nu2 splits the degeneracy of its excited C2E' electronic manifold and exhibits (E [symbol: see text] e) Jahn-Teller (JT) activity. The components of the JT split potential energy surface form conical intersections, and they can also undergo pseudo-Jahn-Teller (PJT) crossings with the X2A1' electronic ground state of B3 via the degenerate nu2 vibrational mode. The impact of the JT and PJT interactions on the nuclear dynamics of B3 in its X2A1'-C2E' electronic states is examined here by establishing a diabatic model Hamiltonian. The parameters of the electronic part of this Hamiltonian are calculated by performing electronic structure calculations and the nuclear dynamics on it is simulated by solving quantum eigenvalue equation. The theoretical results are in good accord with the experimental data. PMID- 16691570 TI - Anion substitution in zinc chalcogenides. AB - Anion substitution effects on the structure and energy of zinc chalcogenides were studied with the semiempirical molecular orbital method MSINDO. Cyclic clusters of different sizes were chosen as model systems. The convergence of the bulk properties of the perfect clusters with increasing cluster size was tested. Single and multiple substitution of oxygen atoms in zinc oxide by sulfur and of sulfur atoms in zinc sulfide by oxygen served to determine the energetics of substitution for these two cases. It was found that the substitution of oxygen by sulfur in ZnO is easier than the substitution of sulfur by oxygen in ZnS in agreement with experimental results. The interaction between two oxygen atoms vs. two selenium atoms in zinc sulfide was investigated. Oscillations of the cluster energy in dependence of the distance between the two doping atoms were observed. These are explained by the relative sites of the doping atoms in the crystal lattice. The magnitude of the oscillations is smaller in ZnS:Se than in ZnS:O, because the difference between the anion radii of S2- and Se2- is smaller than between S2- and O2-. This is also reflected in the band gap. PMID- 16691571 TI - Alterations in alveolar epithelium differentiation and vasculogenesis in lungs of LIF/IGF-I double deficient embryos. AB - Previous studies on double deficient mice for leukemia inhibitory factor (LIF) and insulin-like growth factor I (IGF-I) reported that they died of respiratory failure, with abnormal lung histology and altered expression of pulmonary markers. Here we analyzed prenatal Lif/Igf-I double mutant mouse embryos to characterize LIF and IGF-I cooperative roles in distal lung epithelium and vascular maturation. Lungs of IGF-I-deficient embryos displayed a higher proportion of type II pneumocytes, less differentiated type I pneumocytes, and failure in alveolar capillary remodeling compared to wild type and LIF-deficient mice. Lif/Igf-I double knockout lungs showed aggravated pulmonary hypoplasia, lower airway volume, increased proliferation, and elevated levels of ERK1/2 activation. In addition, their alveoli were collapsed and lined by type II cells. The differentiation of type I cells barely occurred and capillaries remained in the abundant mesenchyme. These results indicate that LIF collaborates with IGF-I in lung alveolar epithelium and vascular maturation. PMID- 16691572 TI - The zebrafish bHLH PAS transcriptional regulator, single-minded 1 (sim1), is required for isotocin cell development. AB - A wide range of physiological and behavioral processes, such as social, sexual, and maternal behaviors, learning and memory, and osmotic homeostasis are influenced by the neurohypophysial peptides oxytocin and vasopressin. Disruptions of these hormone systems have been linked to several neurobehavioral disorders, including autism, Prader-Willi syndrome, affective disorders, and obsessive compulsive disorder. Studies in zebrafish promise to reveal the complex network of regulatory genes and signaling pathways that direct the development of oxytocin- and vasopressin-like neurons, and provide insight into factors involved in brain disorders associated with disruption of these systems. Isotocin, which is homologous to oxytocin, is expressed early, in a simple pattern in the developing zebrafish brain. Single-minded 1 (sim1), a member of the bHLH-PAS family of transcriptional regulatory genes, is required for terminal differentiation of mammalian oxytocin cells and is a master regulator of neurogenesis in Drosophila. Here we show that sim1 is expressed in the zebrafish forebrain and is required for isotocin cell development. The expression pattern of sim1 mRNA in the embryonic forebrain is dynamic and complex, and overlaps with isotocin expression in the preoptic area. We provide evidence that the role of sim1 in zebrafish neuroendocrine cell development is evolutionarily conserved with that of mammals. PMID- 16691573 TI - Fine-needle aspiration of pancreatic serous cystadenoma: cytologic features and diagnostic pitfalls. AB - BACKGROUND: The preoperative diagnosis of pancreatic serous cystadenoma (SCA) is important because as a typically benign tumor it can be treated expectantly, whereas many other cystic tumors require excision. This study examines the cytology, clinical and radiologic features, diagnostic accuracy of fine-needle aspiration (FNA), and potential pitfalls associated with this rare tumor. METHODS: Cytomorphologic features were retrospectively reviewed from 28 FNAs of SCA from 21 patients. FNA biopsies were guided by percutaneous computed tomographic or ultrasonographic imaging in 10 cases and by endoscopic ultrasonographic imaging in 18 cases. Corresponding histology (14 tumors) and clinical/imaging findings were also evaluated. RESULTS: Patients typically presented with upper abdominal discomfort or asymptomatically. Radiologically, a well-demarcated, multiloculated cystic mass involving the pancreatic head or uncinate process was common. Aspirates were sparsely cellular against a clean or granular, proteinaceous background. Tumor cells formed loose clusters or monolayered sheets composed of cuboidal cells with indistinct cell borders and granular or clear cytoplasm that was often stripped from the nucleus. Nuclei were small, round, with fine chromatin and indistinct nucleoli and devoid of mitotic activity. Seven (25%) of the aspirates were initially classified as "consistent with SCA," 6 (21%) as "no malignant cells," 3 (11%) as "nondiagnostic specimen," 3 (11%) as "suspicious for malignancy," 3 (11%) as "rare atypical cells," and 6 (21%) as "probably or consistent with mucinous cystic neoplasm." Features causing diagnostic difficulty were scant cellularity, papillary groups, nuclear atypia, and columnar cells mimicking those of mucinous neoplasms. Gastrointestinal (GI) epithelium and mucin also caused confusion. The detection of intracytoplasmic glycogen (3 of 6 cases) and cyst fluid analysis (2 of 2 cases) showing low viscosity and low or undetectable levels of carcinoembryonic antigen, CA 19.9, and amylase enhanced diagnostic confidence. CONCLUSIONS: Diagnosing SCA by FNA is challenging. Familiarity with its morphologic spectrum, use of ancillary studies, and correlation with clinical/radiologic findings greatly improves diagnostic accuracy. Contaminating GI epithelium and mucin should be distinguished from components of a mucinous neoplasm. PMID- 16691574 TI - A new case of Grange syndrome without cardiac findings. AB - Grange syndrome comprises arterial stenoses with hypertension, brachysyndactyly, bone fragility, learning disability, and cardiac defects. To date, we know of two reported families with five affected individuals. We report on one of the youngest cases, in a third family, a 3-year-old girl with brachysyndactyly, renal artery stenosis with hypertension, and bone fragility. She does not have apparent cardiac disease, suggesting cardiac anomalies may not be an obligatory finding in this syndrome. PMID- 16691575 TI - Using the TBX5 transcription factor to grow and sculpt the heart. AB - TBX5 mutations cause the cardiac and limb defects of the autosomal dominant Holt Oram syndrome (HOS). We have explored the role of the TBX5 transcription factor during cardiogenesis and have elucidated some of its functions in regulating myocardial cell proliferation and proepicardial cell migration. Our identification of TBX5 mutations has enabled us to offer genetic testing for diagnosis of HOS in patients and also to perform preimplantation genetic diagnosis on blastocysts for couples desiring to have a child unaffected by HOS. We hope that our genetic testing approach will serve as a paradigm for mutation screening in other inherited syndromes. PMID- 16691576 TI - Nablus mask-like facial syndrome is caused by a microdeletion of 8q detected by array-based comparative genomic hybridization. AB - In 2000, Teebi reported on a 4-year-old boy with a distinctive pattern of malformation, which he termed the "Nablus mask-like facial syndrome" (OMIM# 608156). Characterization of this syndrome has been difficult because of the paucity of patients described in the medical literature and its unknown etiology and pathogenesis. We present two patients with Nablus mask-like facial syndrome who both display a microdeletion in the 8q21-8q22 region detected by array-based comparative genomic hybridization. Patient 1, a boy, has a distinct facial appearance characterized by severe blepharophimosis, tight-appearing glistening facial skin, sparse and unruly hair, a flat and broad nose, and distinctive ears that are triangular in shape with prominent antihelices. He also demonstrates camptodactyly, contractures, unusual dentition, cryptorchidism, mild developmental delay, and a happy demeanor. Patient 2, a girl with a strikingly similar phenotype, was previously described in a report by Salpietro et al. 2003. She has distinctive ears, dental anomalies, and developmental delay. The etiology of her pattern of malformation was not identified at that time. Although high resolution chromosome and subtelomeric FISH analyses were normal, array-based comparative genomic hybridization revealed an approximately 4 Mb deletion involving the 8q21.3-8q22.1 region in both patients. This region encompasses a number of genes that may contribute to this unique phenotype. These results demonstrate a chromosomal microdeletion as the etiology of Nablus mask-like facial syndrome and emphasize the diagnostic utility of array-based comparative genomic hybridization in the evaluation of multiple malformation syndromes of previously unrecognized causation. PMID- 16691577 TI - Twelve-year mortality results of a randomized trial of 2 versus 5 years of adjuvant tamoxifen for postmenopausal early-stage breast carcinoma patients (SITAM 01). PMID- 16691578 TI - A novel RSK2 (RPS6KA3) gene mutation associated with abnormal brain MRI findings in a family with Coffin-Lowry syndrome. AB - Coffin-Lowry syndrome (CLS) is an X-linked mental retardation syndrome caused by defects in the RSK2 gene. We have identified a CLS family with four patients in two generations. The patients in this family, a mother and her three children (a male and two females), all have severe mental retardation with the typical CLS phenotype. In addition, brain MRI studies on the three siblings revealed abnormalities in deep subcortical white matter, thinning of the corpus callosum, hypoplastic cerebellar vermis, and asymmetry of the lateral ventricles. The degree of severity of the MRI findings correlated with the severity of mental retardation in the patients. Extensive mutation screening was performed on the entire RSK2 gene in this family. Twenty-two exons including the intron/exon junctions were amplified by PCR and subsequently sequenced on both strands. A novel mutation, a two-nucleotide insertion (298 ins TG), was identified. The insertion creates a stop codon at codon 100, resulting in a 99 amino acid truncated RSK2 protein. All patients tested have the same mutation, and no other mutation could be found in the RSK2 gene from the proband. The mutation was confirmed by PCR/RFLP. X-chromosome inactivation assay on the female patients revealed significant skewing toward inactivation of the normal RSK2 allele. Thus, this novel mutation is likely to be responsible for the unusual clinical presentation in this family, which includes full phenotypic expression in females and unique brain MRI abnormalities. The pathological function of the mutation and genotype/phenotype correlation between the mutation and this unusual clinical presentation await further clarification. PMID- 16691580 TI - Prospective evaluation of the prognostic relevance of molecular staging for urothelial carcinoma. AB - BACKGROUND: Nearly 50% of urothelial carcinoma patients with lymph node-negative invasive cancers recur after radical surgery. In many cases, occult local or lymph node disease may be present but undetectable by current approaches. Reverse transcriptase polymerase chain reaction (RT-PCR)-detectable mRNA of Uroplakin II (UPII), a urothelial-specific gene mRNA, was evaluated in perivesical and lymph node samples removed at radical surgery as a predictor of clinical recurrence. METHODS: From November 1999 to August 2002, 46 patients with cTa-T4N0M0 urothelial bladder cancer enrolled in a prospective clinical trial and underwent radical cystectomy and pelvic lymphadenectomy. RT-PCR for UPII was performed on biopsies of the external surface of the bladder specimen and lymph nodes. Results were compared with conventional pathology. Patients were followed every 6 months for tumor recurrence. RESULTS: Pathologically node-negative patients had a UPII RT-PCR perivesical positivity of 27% and a lymph node positivity rate of 33%. All 22 UPII RT-PCR node-negative patients were pathologically node-negative and all 13 with pathologically positive nodes had positive UPII RT-PCR lymph node signals. In all, 46% of UPII RT-PCR lymph node-positive patients were pathologically node-negative and 5% of pathologically node-negative/UPII RT-PCR node-negative patients had disease recurrence, whereas 91% of pathologically node negative/UPII RT-PCR node-positive patients (P < .001) recurred. UPII RT-PCR node positivity was a significant predictor of tumor recurrence in multivariate analysis CONCLUSIONS: Molecular determination of lymph node metastases by UPII RT PCR node positivity apparently identifies patients with a poor prognosis and may be more predictive of disease recurrence than conventional pathologic analysis. PMID- 16691581 TI - Metaphyseal dysplasia of Braun-Tinschert type: report of a Japanese girl. AB - We report on a 7-year-old Japanese girl with metaphyseal dysplasia (MD) of Braun Tinschert type, a recently recognized, autosomal dominant sclerosing bone dysplasia. All individuals with the disorder from four families in the literature originated from a small town in Bohemia or its vicinity. The occurrence of the disorder in a Japanese girl indicates that it is not restricted to Germans. The radiographic hallmarks of the disorder include metaphyseal undermodeling (Erlenmeyer-flask deformity); osteosclerosis of the chondroosseous junctions, metaphyseal cortices, and epiphyseal margins; and exostosis-like bone excrescences at the metaphyseal-diaphyseal junctions. In the girl we described, the latter two findings were conspicuous at age 4 years, but became less prominent with increasing age. The metaphyseal trabeculae were somewhat coarse. The humeri exhibited varus deformity, and the ulnae and fibulae mild bowing. The mean bone mineral density of the lumbar spine was lower than that of age-matched controls. The patient exhibited premature loss of primary teeth, likely to be a sign of increased periodontal bone resorption. Markers of bone formation and resorption were both increased, an indication of a high rate of bone turnover. PMID- 16691582 TI - Hydrocephaly, penoscrotal transposition, and digital anomalies associated with de novo pseudodicentric rearranged chromosome 13 characterized by classical cytogenetic methods and mBAND analysis. AB - We report on a patient with hydrocephaly, penoscrotal transposition, oligodactyly, and minor anomalies. Comprehensive cytogenetic studies involving both classical cytogenetic methods and mBAND analysis were used to show a stable dicentric rearranged chromosome 13 that result in a 46,XY,psu dic(13;13)(13pter - > 13q32::13q11 --> 13pter) de novo karyotype. This aberration probably arose as a consequence of unequal sister chromatid breakage repair events. This report is the first to describe all of the most severe features associated with partial monosomy in one patient. Moreover, the delineation of monosomy 13q32 --> qter in this patient facilitates identification of the developmental genes responsible for the clinical features within this region of chromosome 13. PMID- 16691583 TI - Genetic drift: Unexpected resiliency. PMID- 16691584 TI - Comprehensive screening of multiple epiphyseal dysplasia mutations in Japanese population. AB - Multiple epiphyseal dysplasia (MED) is among the most genetically heterogeneous skeletal dysplasias. Six genes involved in MED, COMP, MATN3, COL9A1, COL9A2, COL9A3, and DTDST have been identified; however, the presence of additional disease genes has been reported, and the detection rate for mutations in known genes accounts for no more than 50% of patients with MED in Western populations. Here, we screened the six known disease genes in 35 consecutive Japanese MED patients. We analyzed the entire coding region of each gene, along with flanking intron-exon junctions, by direct sequencing. A total of 19 mutations were identified in COMP, MATN3, COL9A2, COL9A3, and DTDST. The detection rate for known mutations was higher in this study than in previous reports, and we identified a substantially different spectrum of mutations. Mutations in MATN3 were more prevalent among these Japanese patients, whereas no DTDST mutations were detected. Most of the mutations were localized within specific regions of each gene: COMP mutations were found in the calmodulin-like repeat domains; MATN3 mutations in the von Willebrand factor type A domain; and type IX collagen gene mutations occurred in the third collagenous domains. Based on the integration of clinical and genetic information, we propose an algorithm for detecting mutations in Japanese MED patients. Our study further supports the existence of additional MED gene(s). PMID- 16691585 TI - The use of inappropriate, demeaning, and pejorative terminology in gene nomenclature: a comment on Feingold. PMID- 16691586 TI - Periventricular nodular heterotopia and Williams syndrome. AB - We report here on the first case of a child with bilateral periventricular nodular heterotopia (PNH) and Williams syndrome. Fluorescent in situ hybridization (FISH) analyses demonstrated a deletion of the elastin gene in the Williams syndrome critical region (WSCR). Further mapping by loss of heterozygosity analysis both by microsatellite marker and SNP profiling demonstrated a 1.5 Mb deletion beyond the telomeric end of the typical WSCR. No mutations were identified in the X-linked filamin-A gene (the most common cause of PNH). These findings suggest another dominant PNH disorder along chromosome 7q11.23. PMID- 16691587 TI - Clinical and molecular characterization of individuals with 18p deletion: a genotype-phenotype correlation. AB - The deletion 18p syndrome is one of the most common chromosome abnormalities. The medical problems are mental and postnatal growth retardation, and sometimes malformations of the heart and brain. The individuals have some typical features, which might be easy to overlook and which are: ptosis, strabismus, hypertelorism, broad flat nose, micrognathia, big and low set ears. The aims of present study were to clinically and molecularly characterize the syndrome further in seven subjects with de novo 18p deletions and to perform genotype-phenotype correlation. All seven subjects had terminal deletions and no interstitial deletion was observed with subtelomeric FISH analyses. To define the extent of the 18p deletions and the parental origin of the deletion microsatellite- and FISH analyses were performed on genomic DNA and on lymphoblastoid cell lines of the study participants. Totally 19 chromosomes, 18 specific polymorphic microsatellite markers, and 5 BAC clones were used. The results revealed that the deletions were located in the centromeric region at 18p11.1 in four of the seven subjects. In the remaining three the breakpoints were located distal to 18p11.1 (18p11.21-p11.22). Four of the individuals had a paternal and three a maternal origin of the deletion. Genotype-phenotype correlation of the seven subjects suggests a correlation between the extent of the deleted region and the mental development. All the four children with a deletion in the centromeric region at 18p11.1 had a mental retardation (MR). Two of the three children with a more distal breakpoint (distal 18p11.21) had a normal mental development and one had a border-line mental retardation. There might be a critical region for the mental retardation located between 18p11.1 and 18p11.21. The children with a breakpoint at 18p11.1 had all a broad face, which was observed in only one of those with a more distal breakpoint, otherwise no genotype-phenotype correlation of the features was observed. PMID- 16691588 TI - 3' UTR polymorphism of the serotonin transporter gene and sudden infant death syndrome: haplotype analysis. PMID- 16691589 TI - New cases of Bohring-Opitz syndrome, update, and critical review of the literature. AB - We report on four additional unrelated cases of Bohring-Opitz syndrome with the highly characteristic phenotype of facial anomalies including bulging forehead over the metopic suture, frontal nevus flammeus, exophthalmos, hypertelorism, upslanting palpebral fissures, and cleft lip and/or palate, as well as flexion deformities of the upper limbs, multiple other anomalies, and severe failure to thrive. We also update the clinical outcome of the patients reported in the original article by Bohring et al. [Am J Med Genet 85:438-446] and critically review the subsequently published cases considered to have Bohring-Opitz syndrome. PMID- 16691590 TI - Craniofacial dyssynostosis in two boys with apparently normal cognitive development. PMID- 16691591 TI - Report of a del22q11 in a patient with Mayer-Rokitansky-Kuster-Hauser (MRKH) anomaly and exclusion of WNT-4, RAR-gamma, and RXR-alpha as major genes determining MRKH anomaly in a study of 25 affected women. PMID- 16691592 TI - PHOX2B analysis in non-syndromic neuroblastoma cases shows novel mutations and genotype-phenotype associations. AB - Neuroblastoma (NB) is an embryonal tumor originating from neural crest cells and is one of the most common solid tumors of childhood. Recently, constitutional mutations in PHOX2B have been shown to confer an increased risk of NB. To date, mutations predisposing to neural crest tumors have been reported in 20 individuals from 16 families. These families included additional clinical features such as Hirschsprung (HSCR) disease or congenital central hypoventilation syndrome, either in the index case or relatives. The contribution of PHOX2B mutations to NB cases without additional features is unclear. To address this we sequenced PHOX2B in constitutional DNA from 86 individuals with non-syndromic NB (4 cases had a family history of NB). We identified two mutations, 600delC, a frameshift mutation in an individual with isolated, unifocal NB and G197D, a missense mutation that was present in a family with multiple individuals with NB but no evidence of autonomic dysfunction. These data demonstrate that PHOX2B mutations are a rare cause of non-syndromic NB. The mutations we identified are outside the domains typically mutated in PHOX2B syndromes. This provides further evidence that the underlying PHOX2B mutational mechanism influences tumor risk and suggests that the position of missense mutations may influence the resulting phenotype. PMID- 16691593 TI - Covariation of corticospinal efficiency and silent period in motoneuron diseases. AB - For a better understanding of the changes affecting the cortically induced silent period (SP) in motoneuron disease, the excitatory and inhibitory effects of transcranial magnetic stimulation were explored repeatedly in 8 patients with amyotrophic lateral sclerosis (ALS), 3 patients with Kennedy's disease (KD), and 10 healthy subjects. In KD, the background electromyogram (EMG) and the motor evoked potential (MEP) area were both enhanced. However, neither the corticospinal efficiency (MEP gain, the ratio between MEP and background EMG) nor the duration of the SP differed from healthy subjects. In ALS patients, the MEP gain and the SP duration decreased conspicuously with time. We conclude that use of the MEP gain improves detection of corticospinal dysfunction in ALS patients. Part of the SP shortening in ALS seems to reflect the reduced activation of cortical or spinal inhibitory networks by the abnormal corticospinal pathway. PMID- 16691594 TI - Autosomal dominant syndrome resembling Coffin-Siris syndrome. AB - Coffin-Siris syndrome is a multiple congenital anomaly/mental retardation syndrome with phenotypic variability [OMIM 135900]. The diagnosis is based solely on clinical findings, as there is currently no molecular, biochemical, or cytogenetic analysis available to confirm a diagnosis. Although typically described as an autosomal recessive disorder, autosomal dominant inheritance has also been infrequently reported. We describe a mother and her two daughters who all have features that resemble Coffin-Siris syndrome. However, this is not a completely convincing diagnosis given that hypertelorism is not a feature of Coffin-Siris syndrome and the family is relatively mildly affected. Yet, this family provides further evidence of an autosomal dominant mode of inheritance for a likely variant of Coffin-Siris syndrome (at least in some families). In addition, Sibling 1 had premature thelarche. She is the second reported individual within the spectrum of Coffin-Siris syndrome to have premature thelarche, indicating that it may be a rare clinical feature. PMID- 16691595 TI - Trigonocephaly in a boy with paternally inherited deletion 22q11.2 syndrome. AB - Deletion 22q11.2 syndrome is a well-known contiguous gene syndrome, for which the list of findings is extensive and varies from patient to patient. We encountered a unique patient who had a familial 3-Mb deletion 22q11.2 associated with trigonocephaly derived from craniosynostosis of the metopic suture. Almost all the symptoms of the patient, including polymicrogyria, microcephaly, facial abnormalities, internal anomalies, seizures, and mental retardation, were compatible with deletion 22q11.2 syndrome, except for synostosis of the metopic suture. This is the first report of a relationship between deletion 22q11.2 syndrome and trigonocephaly. Craniosynostosis of the metopic suture might be a minor complication of deletion 22q11.2, although coincidental occurrence cannot be ruled out. PMID- 16691596 TI - Signaling networks controlled by the MRN complex and MDC1 during early DNA damage responses. AB - Cells activate complex signaling networks in response to DNA damage. Several proteins and protein complexes are involved in sensing DNA lesions and initiating the DNA damage response networks. The subsequent DNA damage responses, including the initiation of DNA repair pathways, the activation of cell cycle checkpoint controls and the induction of apoptosis, help maintain genomic stability in mammalian systems. Failure to establish the appropriate DNA damage signaling networks results in genomic instability, which is a known causal factor in tumorigenesis. This review will discuss recent progress in the understanding of the mechanisms by which mammalian cells sense DNA lesions and transduce DNA damage signals during early DNA damage responses. PMID- 16691597 TI - Identification of a novel EYA1 mutation presenting in a newborn with laryngomalacia, glossoptosis, retrognathia, and pectus excavatum. PMID- 16691598 TI - Evaluation of SLC35A3 as a candidate gene for human vertebral malformations. PMID- 16691599 TI - Re: Majewski osteodysplastic primordial dwarfism type II (MOPD II) complicated by stroke: Clinical report and review of cerebral vascular anomalies [Brancati et al., 2005: Am J Med Genet 139A:212-215]. PMID- 16691600 TI - The FG syndrome: report of a large Italian series. AB - Initially described as a rare MCA/MR syndrome occurring only in boys, due to a recessive mutation on the X chromosome [Opitz and Kaveggia, 1974], the FG syndrome (FGS) now emerges as a more common disorder also occurring in girls. Based on over 50 reported cases, FGS is associated with developmental delay (especially speech), hypotonia, postnatal onset relative macrocephaly, prominent forehead, frontal hair upsweep, telecanthus, or ocular hypertelorism, thin vermilion border of the upper lip, relatively short fingers with broad thumbs and halluces, persistent fetal fingertip pads, anal anomalies, and/or constipation. Major malformations are rare, and include pyloric stenosis, anal agenesis, cryptorchidism, hypospadias, and congenital heart defects. Abnormal EEGs and seizures have been reported in almost 70% of patients. Brain MRI shows corpus callosum abnormalities associated with dilatation of lateral ventricles and, less frequently, periventricular nodular heterotopias, mild cerebellar defects, and reduced periventricular white matter. Chiari 1 malformation seems to be frequent. The behavior phenotype appears to be characterized by ADHD, and relatively less developed language, fine motor and executive function skills; whereas visual spatial abilities seem to be a relative strength. Five candidate loci are already known but no gene identified. We describe 25 patients referred to the Stella Maris Institute for evaluation of DD/MR, and diagnosed as FGS. They were between 2 and 15 1/2 years at the first observation. High resolution banding, FRAXA/FRAXE DNA analysis, and subtelomere FISH analysis were performed in all of them, and all had normal results. Thirteen patients were followed-up from 6 months to 9 years. Our report focuses on physical, neurological, developmental findings, and natural history of FGS. Experience with our series of patients suggests that the syndrome may be common, and should be routinely considered in the evaluation of children and adolescents with DD/MR. PMID- 16691601 TI - Differential activation and block of peripheral nerve fibers by magnetic fields. AB - The ability to noninvasively and reversibly block conduction in peripheral nerves would have several clinical applications. As an initial step in this direction, we investigated the possibility of magnetically generating and differentially blocking activity in mammalian peripheral nerve fibers in vitro. Compound action potentials at each end of individual explanted phrenic nerves were recorded in response to currents induced at the midpoint of the nerve with an externally placed magnetic coil. Current in the coil was then reversed and the recordings repeated. In all cases, the area under the compound action potential on the virtual anode side of the magnetic stimulus was reduced (mean of 18.2 +/- 8.8%) in comparison to the area on the virtual cathode side. This indicates that peripheral nerve activity can be differentially induced by magnetic stimulation. Extension of this effect to the point of generating unidirectional action potentials in vivo may prove clinically useful in a number of contexts, such as reducing contractures secondary to spasticity and generating magnetically induced anesthesia in limbs. Further investigations of this effect seem warranted. PMID- 16691602 TI - Skewed X-chromosome inactivation is not associated with premature ovarian failure in a large cohort of Italian patients. PMID- 16691603 TI - Central fatigue in continuous and intermittent contractions of triceps brachii. AB - Conflicting results have been found across studies concerning the effect of rest periods on the development of central fatigue during prolonged muscle activity. Thus, the aim of the present study was to assess differences in the development of central fatigue between continuous and intermittent elbow extension fatigue tasks in the same subjects. Force and electromyographic data were collected on eight healthy volunteers. The ability to maximally activate the triceps brachii muscle was assessed by delivering trains of electrical stimulation during maximal voluntary efforts. This was done before, during, and after three fatigue tasks involving a maximal contraction in elbow extension. One short-duration ( approximately 55-s) and two long-duration (3-min) fatigue tasks were performed by all subjects on separate sessions. One 3-min task was intermittent (5-s rests every 30 s) and the other was continuous. The main findings were that the development and extent of central fatigue were task-dependent, with a greater decrease in the ability to maximally activate triceps brachii observed for the 3 min continuous task. Also, the voluntary activation (VA) ratio was found to be a more sensitive index of central fatigue than the central activation ratio (CAR). These results suggest that, when assessing central fatigue in patients, conclusions may vary depending on the continuous/intermittent nature of the task performed and the estimate of voluntary activation used. PMID- 16691604 TI - Comparison of continuous and intermittent vibration effects on rat-tail artery and nerve. AB - Hand-transmitted vibration from powered-tools can cause peripheral vasospasm and neuropathy. A rat-tail model was used to investigate whether the pattern of vibration influenced the type and severity of tissue damage. The tails of awake rats were vibrated continuously or intermittently for a total of 4 hours at 60 HZ, 49 m/s(2). Nerves and arteries were harvested immediately or 24 hours after treatment. Tails subjected to intermittent vibration showed transiently increased sensitivity to thermal stimuli. Intermittent vibration caused the most nerve injury immediately and 24 hours after vibration. Continuous vibration invoked a persistent reduction in vascular lumen size. Compared to epinephrine-induced transient vacuolation in vascular smooth muscle cells, both continuous and intermittent vibration caused greater persistence of vacuoles, indicating a vibration-induced pathological process. All vibration groups exhibited elevated nitrotyrosine immunoreactivity indicative of free-radical damage. Pattern of vibration exposure may exert a major influence on the type of vibration injury. PMID- 16691605 TI - Sural neuropathy: etiologies and predisposing factors. AB - Isolated sural neuropathy is an uncommon diagnosis. We identified 36 patients with isolated sural neuropathy. Sixteen had various forms of ankle trauma, in three of whom the associated sural neuropathies developed following medical intervention. Three patients developed sural neuropathy associated with vasculitis, and there were single patients with schwannoma and ganglionic cyst. In patients without a history of trauma, structural causes, such as schwannoma or ganglionic cysts and vasculitis, should be considered and managed as appropriate. PMID- 16691606 TI - Pulmonary function abnormalities and airway irritation symptoms of metal fumes exposure on automobile spot welders. AB - BACKGROUND: Spot or resistance welding has been considered less hazardous than other types of welding. Automobile manufacturing is a major industry in Taiwan. Spot and arc welding are common processes in this industry. The respiratory effects on automobile spot welders exposed to metal fumes are investigated. METHODS: The cohort consisted of 41 male auto-body spot welders, 76 male arc welders, 71 male office workers, and 59 assemblers without welding exposure. Inductivity Coupled Plasma Mass Spectrophotometer (ICP-MS) was applied to detect metals' (zinc, copper, nickel) levels in the post-shift urine samples. Demographic data, work history, smoking status, and respiratory tract irritation symptoms were gathered by a standard self-administered questionnaire. Pulmonary function tests were also performed. RESULTS: There were significantly higher values for average urine metals' (zinc, copper, nickel) levels in spot welders and arc welders than in the non-welding controls. There were 4 out of 23 (17.4%) abnormal forced vital capacity (FVC) among the high-exposed spot welders, 2 out of 18 (11.1%) among the low-exposed spot welders, and 6 out of 130 (4.6%) non welding-exposed workers. There was a significant linear trend between spot welding exposure and the prevalence of restrictive airway abnormalities (P = 0.036) after adjusting for other factors. There were 9 out of 23 (39.1%) abnormal peak expiratory flow rate (PEFR) among high-exposed spot welders, 5 out of 18 (27.8%) among the low-exposed spot welders, and 28 out of 130 (21.5%) non-welding exposed workers. There was a borderline significant linear trend between spot welding exposure and the prevalence of obstructive lung function abnormalities (P = 0.084) after adjusting for other factors. There was also a significant dose response relationship of airway irritation symptoms (cough, phlegm, chronic bronchitis) among the spot welders. Arc welders with high exposure status also had a significant risk of obstructive lung abnormalities (PEFR reduction). There was also a significant dose-response relationship of airway irritation symptoms (cough, phlegm, chest tightness, and chronic bronchitis) among the arc welders. CONCLUSIONS: These findings suggest that restrictive and obstructive lung abnormalities, and airway irritation symptoms are associated with spot and arc welding exposures. PMID- 16691607 TI - Adverse pregnancy outcomes in offspring of fathers working in biomedical research laboratories. AB - BACKGROUND: Laboratory work may constitute a possible health hazard for workers as well as for their offspring, and involves a wide range of exposures, such as organic solvents, carcinogenic agents, ionizing radiation, and/or microbiological agents. Adverse pregnancy outcomes in the offspring of male employees in biomedical research laboratories are examined. METHODS: Offspring to males employed 1970-1989 at four Swedish universities were identified via the Medical Birth Register (MBR), along with other pregnancy parameters. Offspring of fathers with laboratory work (n = 2,281) is considered exposed, and of non-laboratory employees unexposed (n = 1,909). Exposure data were obtained by questionnaires to research group leaders. Logistic regression analysis estimated odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: Paternal laboratory work in general showed no statistically significant increased ORs concerning birth weight and/or gestational age, but work specifically with radioactive isotopes gave OR 1.8 (CI 1.0-3.2) for high birth weight and a relative risk of 1.2 (CI 1.0-1.4) for sex ratio (male/female). CONCLUSIONS: There was no clear association between periconceptional paternal laboratory work and adverse reproductive outcomes, but use of radioactive isotopes showed increased OR for high birth weight in offspring. PMID- 16691608 TI - Ulcerative colitis reactivation after mercury vapor inhalation. AB - BACKGROUND: After exposure to mercury vapor at three consecutive 10-month intervals, an electrician in an electroplating plant had flare-ups of ulcerative colitis within 24 hr, that subsided in several days, then returned upon re exposure 10 months later. METHODS: The patient and his workplace were both evaluated for mercury exposure. In addition to workplace inspection, both personal and area monitoring for environmental mercury was performed, using both multiple mercury diffusion badges and direct (instantaneous) readings, during maintenance of mercury-filled electrical blocks. RESULTS: Eight-hour time weighted average (TWA) mercury vapor exposure was measured at 0.41 mg/m3 (ACGIH and NIOSH recommended TWA = 0.025 mg/m3; OSHA permissible exposure limit -0.1 mg/m3) for 5 years since stopping overexposure to mercury, the patient remained symptom free in clinical remission. CONCLUSIONS: In a patient with chronic ulcerative colitis in remission, occupational exposure to mercury vapor led to episodes of disease reactivation. PMID- 16691609 TI - Longitudinal relationship of work hours, mandatory overtime, and on-call to musculoskeletal problems in nurses. AB - BACKGROUND: Nurses are at very high risk for work-related musculoskeletal injury/disorders (MSD) with low back pain/injury being the most frequently occurring MSD. Nurses are also likely to work extended schedules (long hours, on call, mandatory overtime, working on days off). The purpose of this study was to examine the relationship of extended work schedules in nurses to MSD. METHODS: Using a longitudinal, three wave survey of 2,617 registered nurses, Wave 1 work schedule data were related to neck, shoulder, and back (MSD) cases occurring in Waves 2 or 3. RESULTS: Schedule characteristics increasing MSD risk included 13+ hour/days, off-shifts, weekend work, work during time off (while sick, on days off, without breaks), and overtime/on-call. These increases in risk were not explained by psychological demands, but were largely explained by physical demands. CONCLUSIONS: Adverse schedules are significantly related to nurse MSD. Healthier schedules, less overtime, and reducing work on days off would minimize risk and recovery time. PMID- 16691610 TI - Exposure to silica and risk of ANCA-associated vasculitis. AB - BACKGROUND: Exposure to silica dust is considered to be one of etiological factors of antineutrophil cytoplasmic antibodies (ANCA) -associated vasculitis (AAV). METHODS: Subjects exposed to silica dust in Central Bohemia and followed in the Department of Occupational Medicine, Charles University, were selected for study. A group of 86 men exposed to SiO2 for at least 5 years were examined. The association between occupational exposure to silica dust and ANCA positivity is analyzed. RESULTS: The subjects had a mean age of 66.7 years, and mean exposure to silica of 22.3 years. ANCA were detected significantly more frequently in patients group (17.1%; P-ANCA 18x, C-ANCA 3x) than in controls (n = 28, mean age 64.2 years, P-ANCA 1x, i.e., 3.6%). ANCA positivity was found less frequently (7.1%) in the group with history of SiO2 exposure without signs of pronounced silicosis, than in the group with simple (30.3%) or complicated silicosis (36.0%). Odds ratio for ANCA positivity and relative risk estimate in patients with silicosis were highly significant. Among possible predictor factors for ANCA positivity, silicosis and tuberculosis were relevant. No typical AAV was present among the patients. CONCLUSION: Long-term silica exposure may be one of the exogenous factors contributing to ANCA production, however, silica exposure alone, without typical silicosis, was not associated with ANCA positivity. PMID- 16691611 TI - Metal and non-metal miners' exposure to crystalline silica, 1998-2002. AB - BACKGROUND: Crystalline silica is well known to cause silicosis and other diseases. Exposure is common in the mining industry and consequently, the US Mine Safety and Health Administration (MSHA) evaluates miners exposure to silica to determine compliance with its exposure limit. METHODS: MSHA exposure measurements were obtained for the 5-year period from 1998 to 2002 and average exposure was calculated classified by occupation and by mine. Evaluation criteria were whether average values exceeded MSHA's permissible exposure limit or the limit recommended by the National Institute for Occupational Safety and Health (NIOSH), whether there was a risk of exposure to freshly fractured silica, and whether there was a risk of a high rate of exposure to silica. RESULTS: Miners in certain jobs are exposed to silica above permissible and recommended exposure limits. Some miners may also be exposed at a high rate or to freshly fractured silica. CONCLUSIONS: Known dust control methods should be implemented and regular medical surveillance should be provided. PMID- 16691612 TI - Capturing cases in workers' compensation databases: the example of neck pain. AB - BACKGROUND: There is a need to more accurately enumerate workers with musculoskeletal injuries who make lost-time claims to workers compensation boards. The objective of this study is to develop an approach to more accurately enumerate these workers. METHODS: Lost-time claims to the Ontario Workplace Safety & Insurance Board (WSIB) were reviewed. Using neck pain as an example, nature of injury and part of body codes were identified to classify cases. Claims of a random sample of 434 claimants were reviewed. The proportion of claimants classified as having neck pain was computed. RESULTS: The proportion of claimants classified with soft-tissue injuries to the neck varied from 0.88 for codes including "neck/cervical region," 0.69 for "back region" to 0.05 for those coded as "shoulder/upper arm." CONCLUSIONS: Restricting the enumeration of injuries to specific part of body codes can lead to a gross underestimation of the magnitude of soft-tissue disorders in epidemiological studies using workers' compensation data. The proposed approach leads to more accurate enumeration. PMID- 16691613 TI - Mortality in a Swedish rubber tire manufacturing plant: occupational risks or an "unhealthy worker" effect? AB - BACKGROUND: In this cohort study a classification of 12 work categories, with specified common exposures, is used to evaluate the mortality among workers in a Swedish rubber tire plant. METHODS: Mortality among the cohort members is compared with expected values from national rates. Standardized mortality ratios (SMR) were calculated for the total cohort, for sub-cohorts and with the inclusion of a latency requirement. RESULTS: Male workers had increased risks for total mortality, circulatory death, respiratory death, mental disorders, epilepsy, injuries/poisoning, and for alcohol-related death. Many risks were noted among workers exposed to dust, fumes, or vapor. No trend in risk with increasing duration of employment was seen. DISCUSSION: Some established risks for rubber workers could be related to some specified work categories. The highest risks were noted among workers having a very short duration of employment indicating a possible "unhealthy worker effect" associated with life-style factors or to tasks offered to unskilled workers. PMID- 16691614 TI - Respiratory health in Turkish asbestos cement workers: the role of environmental exposure. AB - AIM: Benign and malignant pleural and lung diseases due to environmental asbestos exposure constitute an important health problem in Turkey. The country has widespread natural deposits of asbestos in rural parts of central and eastern regions. Few data exists about the respiratory health effects of occupational asbestos exposure in Turkey. A cross-sectional study was conducted to investigate respiratory health effects of occupational asbestos exposure and the contribution of environmental asbestos exposure. METHODS: Investigations included asbestos dust measurements in the workplace and application of an interviewer-administered questionnaire, a standard posteroanterior chest X-ray and spirometry. Information on birthplace of the workers was obtained in 406 workers and used to identify environmental exposure to asbestos, through a map of geographic locations with known asbestos exposure. RESULTS: Asbestos dust concentration in the ambient air of the work sites (fiber/ml) ranged between 0.2 and 0.76 (mean: 0.25, median: 0.22). Environmental exposure to asbestos was determined in 24.4% of the workers. After the adjustment for age, smoking, occupational asbestos exposure, and potential risk factors environmental asbestos exposure was associated with small irregular opacities grade > or = 1/0 (44.2% vs. 26.6%, P < 0.01), FVC% (97.8 vs. 104.5, P < 0.0001), and FEV1% (92.4 vs. 99.9, P < .0001). Occupational exposure to asbestos was associated with small irregular opacities grade > or = 1/0 (OR: 2.0, 95% CI: 1.3-3.1, per 1 unit increase in the natural logarithm of fiber/ml) and FEV1/FVC% (beta: 1.1, SEM: 0.54; P < 0.05, per 1 unit increase in the natural logarithm of fiber/ml). CONCLUSIONS: Environmental exposure to asbestos could increase the risk of asbestosis and lung function impairment in workers occupationally exposed to asbestos, independent from occupational exposure and smoking. PMID- 16691615 TI - Attenuation of retinal vascular development and neovascularization in transgenic mice over-expressing thrombospondin-1 in the lens. AB - Thrombospondin-1 (TSP1) is an endogenous inhibitor of angiogenesis and induces endothelial cell (EC) apoptosis. To study the role TSP1 plays during vascular development and neovascularization, we assessed the effects of ectopic TSP1 expression in the lens on retinal vascularization in transgenic mice. The TSP1 over-expressing mice showed abnormalities in the development of retinal vasculature. There was a dramatic decrease in the density of superficial and deep vascular plexuses of the retina in transgenic mice. The retinal vessels in TSP1 transgenic mice also appeared nonuniform and abnormal in maturation. We detected an increase in the number of EC undergoing apoptosis, which was compensated, in part, by an increase in cell proliferation in retinal vasculature of TSP1 transgenic mice. The TSP1 transgenic mice also exhibited increased levels of vessel obliteration and a limited preretinal neovascularization during oxygen induced ischemic retinopathy (OIR). Our results indicate increased expression of TSP1 attenuates normal retinal vascularization and preretinal neovascularization during OIR. Therefore, modulation of TSP1 expression may provide an effective mechanism for regulation of ocular angiogenesis. PMID- 16691616 TI - Gum mastic inhibits the expression and function of the androgen receptor in prostate cancer cells. AB - Accumulating evidence suggests that the androgen receptor (AR) may play an important role in the development and progression of prostate cancer. To find new, useful compounds that effectively may attenuate the function of AR in prostate cancer cells, the authors investigated the effect of gum mastic, a natural resin, on AR activity. An androgen-responsive prostate cancer cell line LNCaP was used as a model for this study. Gene transfer, reverse transcriptase polymerase chain reaction analysis, electrophoretic mobility shift assay, and Western blot analysis were used to test the effect of gum mastic on the expression and function of the AR. To demonstrate the inhibitory effect of gum mastic on the function of the AR, the expression of androgen-regulated genes, including prostate-specific antigen (PSA), human kallikrein 2 (hK2), and NKX3.1 were measured. In addition, transient transfection assays with the PSA promoter and the AR promoter also were used to test the effects of mastic. The results showed that gum mastic inhibited the expression of the AR at the transcriptional level, resulting in the down-regulation of both AR messenger RNA and protein levels. Therefore, the function of the AR was inhibited, as reflected by the reduced expression of NKX3.1 and PSA and by androgen-stimulated growth. Because gum mastic exhibited a strong in vitro potency to attenuate the expression and function of the AR, further investigation will be required to determine whether this naturally occurring substance has in vivo potency to inhibit prostate cancer development. PMID- 16691617 TI - Supraclavicular radiation for breast cancer does not increase the 10-year risk of stroke. AB - BACKGROUND: Modern radiation techniques, which limit the radiation dose to the heart during treatment for breast cancer, have greatly reduced the risk of radiation-induced cardiac injury. However, the risk of radiation damage to the carotid artery, which is often incidentally included in the supraclavicular radiation field for breast cancer treatment, is not routinely examined, and the technique used to treat this field has not changed significantly from early radiation trials. The purpose of the current study was to compare the incidence of hospitalization for stroke among women with breast cancer treated with supraclavicular radiation with those who received radiation therapy to the breast but not the supraclavicular fossa. METHODS: Data from the Surveillance, Epidemiology, and End Results (SEER)-Medicare database for 5752 women who were diagnosed with American Joint Committee on Cancer (AJCC) Stage I-III nonmetastatic breast cancer between 1988 and 1997 were analyzed. Women included were age > or = 66 years, had known lymph node (LN) status, had tumors measuring < or = 5 cm, underwent breast surgery, and received adjuvant radiation therapy (with or without supraclavicular irradiation). Patients with < 5 years of follow up were excluded because events in the first 5 years after radiation were unlikely to be radiation induced. A Cox proportional hazards model was used to compare patients with 0 positive LNs (surrogate group for no supraclavicular radiation, n = 5281) with patients with > 4 positive LNs (surrogate for supraclavicular radiation group, n = 471) for the endpoint of hospitalization for stroke. Time-to-event curves were calculated using the conditional Kaplan-Meier method. RESULTS: The median follow-up for the 0 and 4+ LN cohorts were 92 months and 90 months, respectively (minimum of 60 months). The 10-year and 15-year actuarial freedom from hospitalization for stroke was 91% (0 LN) versus 89.5% (4+ LN) and 79% (0 LN) versus 81.6% (4+ LN), respectively (P = .28). Estrogen receptor status was balanced between the 2 cohorts. As expected, the 4+ cohort had more advanced tumors, higher stage, larger tumor size, and higher grade (P < .0001). In multivariate analysis including LN group, year of diagnosis, age, race, type of surgery, stage, tumor size, grade, estrogen receptor status, and Charlson comorbidity score, only increased age (hazard ratio [HR] for ages 70-74 years, 1.6; HR for ages 75-79 years, 2.1; and HR for age 80 + years, 2.7) and increasing comorbidity score were predictive of an increased risk of hospitalization for stroke. CONCLUSIONS: Although patients with nonbreast malignancies treated with higher doses to the carotid arteries have been shown to have an increased risk of carotid injury, no evidence was found that radiation to the carotid delivered during supraclavicular irradiation for breast cancer increases the risk of hospitalization for stroke. PMID- 16691618 TI - Dose escalation of oral vinorelbine in combination with estramustine in hormone refractory adenocarcinoma of the prostate. AB - BACKGROUND: The primary objective of the current study was to identify the tolerable dose level of oral vinorelbine when given in combination with estramustine to men with hormone-refractory prostate cancer (HRPC). The secondary objectives were to describe the toxicities of the combined regimen in patients with HRPC and to estimate the efficacy of oral vinorelbine in combination with estramustine based on the prostate-specific antigen (PSA) response. METHODS: Thirty-three patients with HRPC were treated on a 28-day cycle with estramustine at a dose of 140 mg orally 3 times a day on Days 1-3 and 8-10. Vinorelbine was given orally on Days 2 and 9. The initial dose of vinorelbine was 50 mg/m2 and was escalated to 70 mg/m2 using the time-to-event continual reassessment method. RESULTS: Three of 17 patients experienced dose-limiting toxicity at the 70 mg/m2 dose level of oral vinorelbine. One patient experienced dose-limiting toxicity at a dose of 60 mg/m2 and no dose-limitig toxicities were reported at the 50 mg/m2 dose. The overall response rate by > or = 50% reduction in PSA was 17.2%, (95% confidence interval, 5.9-35.8%). CONCLUSIONS: Oral vinorelbine at doses of 70 mg/m2 may be safely combined with estramustine. The combination appears to have modest activity in men with advanced prostate cancer. The trial design employed the time-to-event continual reassessment method, which potentially allows for rapid accrual, a more complete assessment of toxicities, and a larger fraction of patients to be treated at an effective dose. More active regimens are needed to further evaluate the utility of this clinical trial design in patients with prostate cancer. PMID- 16691619 TI - Split-hand/split-foot malformation 3 (SHFM3) at 10q24, development of rapid diagnostic methods and gene expression from the region. AB - Split-hand/split-foot malformation (SHFM, also called ectrodactyly) is a clinically variable and genetically heterogeneous group of limb malformations. Several SHFM loci have been mapped, including SHFM1 (7q21), SHFM2 (Xq26), SHFM3 (10q24), SHFM4 (3q27) and SHFM5 (2q31). To date, mutations in a gene (TP63) have only been identified for SHFM4. SHFM3 has been shown by pulsed-field gel electrophoresis to be caused by an approximately 500 kb DNA rearrangement at 10q24. This region contains a number of candidate genes for SHFM3, though which gene(s) is (are) involved in the pathogenesis of SHFM3 is not known. Our aim in this study was to improve the diagnosis of SHFM3, and to begin to understand which genes are involved in SHFM3. Here we show, using two different techniques, FISH and quantitative PCR that SHFM3 is caused by a minimal 325 kb duplication containing only two genes (BTRC and POLL). The data presented provide improved methods for diagnosis and begin to elucidate the pathogenic mechanism of SHFM3. Expression analysis of 13 candidate genes within and flanking the duplicated region shows that BTRC (present in three copies) and SUFU (present in two copies) are overexpressed in SHFM3 patients compared to controls. Our data suggest that SHFM3 may be caused by overexpression of BTRC and SUFU, both of which are involved in beta-catenin signalling. PMID- 16691620 TI - Recombinant interferon gamma1b immune enhancement in 20 patients with hematologic malignancies and systemic opportunistic infections treated with donor granulocyte transfusions. AB - BACKGROUND: The response to antifungal therapy alone often is suboptimal in patients with cancer who have therapy-refractory neutropenia, and even donor derived granulocyte transfusions (GTX) are not always successful. The authors evaluated the safety and efficacy of immune enhancement using recombinant interferon gamma1b (rIFN-gamma1b) in patients with cancer who received GTX for refractory, systemic, opportunistic infections. METHODS: Twenty recipients of high-dose donor GTX ( approximately 5.5 x 10(10) neutrophils per transfusion) who had received concurrent rIFN-gamma1b between October 2001 and December 2004 were evaluated retrospectively. RESULTS: The median age (+/- standard deviation [SD]) was 45 +/- 17 years. Ten patients (50%) were men, 17 patients (85%) had leukemia, and 3 patients (15%) had myelodysplastic syndrome. The median +/- SD Acute Physiology and Chronic Health Evaluation II score was 15 +/- 4 (range, 7-22). Most patients (n = 18 patients; 90%) had recurrent or refractory cancer. In 6 patients (30%) who received allogeneic hematopoietic stem cell transplantation, GTX plus rIFN-gamma1b was given a median +/- SD of 26 +/- 100 days (range, 12-372 days) after transplantation. Seventeen patients (85%) had neutropenia during GTX therapy. Five patients (25%) had possible invasive fungal infection, 3 patients (15%) had probable invasive fungal infection, and 11 patients (55%) had proven invasive fungal infection. One patient (5%) had refractory Pseudomonas aeruginosa sepsis. Eight patients (40%) received corticosteroids during GTX plus rIFN gamma1b therapy. Patients received a median +/- SD of 8 +/- 7 GTX doses (range, 4 28 doses) and 9 +/- 7 rIFN-gamma1b doses (range, 1-28 doses), for a mean +/- SD cumulative dose (CD) of 400 +/- 2621 microg. Other concomitant cytokines were granulocyte-colony stimulating factor (12 +/- 3 doses; CD, 6720 +/- 4721 microg) in 15 patients (75%) and granulocyte-macrophage-colony stimulating factor (12 +/- 9 doses; CD, 4750 +/- 4410 microg) in 14 patients (70%). Four patients (20%) developed fever, and 2 patients (10%) developed skin rashes. Reversible liver dysfunction (n = 3 patients; 15%) and tachycardia (n = 1 patients; 5%) were considered rIFN-gamma1b-associated adverse reactions; whereas, in 1 patient (5%), transient dyspnea was attributed to GTX. Four weeks after therapy started, 9 patients (45%) had complete or partial resolution of infection; and, in another 3 patients (15%), the invasive fungal infection had become stable. CONCLUSIONS: The current results indicated that no serious adverse events were associated with rIFN-gamma1b immune enhancement in patients with systemic opportunistic infections who received donor GTX therapy. PMID- 16691621 TI - Dedifferentiated central chondrosarcoma. AB - BACKGROUND: The prognosis for patients who develop dedifferentiation of central chondrosarcoma traditionally has been poor. Because not much has been reported about this rare lesion, many uncertainties remain about prognostic factors. METHODS: In this retrospective study, the clinical, radiographic, and histologic features and the treatments in 123 patients from the Rizzoli Institute were reviewed in an attempt to define which factors may be related to outcome in patients with dedifferentiated central chondrosarcoma. RESULTS: Among 123 patients who were included in this study, 109 patients were treated at the Rizzoli Institute, and 14 patients were seen in consultation. There were 66 males and 57 females, and their average age was 59 years. The femur (62 patients), pelvis (28 patients), and humerus (20 patients) were the most common locations. Radiographically, a soft tissue mass was present in 87% of patients, and a bimorphic pattern was appreciated in 53% of patients. Histologically, the cartilaginous component was considered Grade 1 in 63% of patients and Grade 2 in 37% of patients. In most patients, the dedifferentiated component showed the features of an osteosarcoma (92 patients), followed by fibrosarcoma (19 patients), and malignant fibrous histiocytoma (9 patients). For 101 patients, surgery was a component of their definitive management. In 25 patients, surgery was combined with chemotherapy. The 2-year and 5-year survival rates were 34% and 24%, respectively. The median survival was 13 months (95% confidence interval, 9 17 months). CONCLUSIONS: Metastatic disease at diagnosis, malignant fibrous histiocytoma dedifferentiation, and a high percentage of dedifferentiated component were related to poorer outcomes. There was no statistical evidence of any beneficial effect from adjuvant chemotherapy. PMID- 16691622 TI - Pattern of p63 mutations and their phenotypes--update. AB - Heterozygous mutations in the transcription factor gene p63 cause at least six different syndromes with various combinations of ectodermal dysplasia, orofacial clefting and limb malformations. Here we will present an update of mutations in the p63 gene together with a comprehensive overview of the associated clinical features in 227 patients. These data confirm the previously recognized genotype phenotype associations. Moreover, we report that there is a large degree of clinical variability in each of the p63-associated disorders. This is illustrated by the different phenotypes that are seen for the five-hotspot mutations that explain almost 90% of all EEC syndrome patients. PMID- 16691623 TI - Influence of local recurrence on survival in patients with extremity osteosarcoma treated with neoadjuvant chemotherapy: the experience of a single institution with 44 patients. AB - BACKGROUND: Risk factors for local recurrence (LR) after osteosarcoma, such as surgical margins and histologic response to preoperative treatment, have been well documented, whereas the outcome for patients who locally recur has not been well established yet. METHODS: Retrospective analysis of the management and outcome of 44 patients who developed LR after treatment of osteosarcoma of the extremities with neoadjuvant chemotherapy was performed in a single institution between 1983 and 1999. RESULTS: In 24 patients (54.5%), LR was the first sign of recurrence; in 8 patients (18.2%) LR followed systemic recurrence and in 12 patients (27.3%), the 2 events, local and systemic recurrence, were concurrent. Of the 44 patients, 26 with local recurrences were free of disease, but only 5 were long-term event-free survivors, and 39 patients developed further recurrences: 37 died of the tumor and 2 were alive with uncontrolled disease at the time of last follow-up. The 5-year disease-free survival rate after the last recurrence was 15.9%; it was 25.9% for patients who achieved remission and 0% for the others. The only prognostic factor identified for post-LR disease-free survival was the presence of systemic recurrence at the time of diagnosis of LR or before (5-year postrecurrence event-free survival rate of 29.1% for patients without metastases at the time of local recurrence vs. 0% for those with metastases; P = .02). CONCLUSIONS: These results confirm that patients with osteosarcoma of the extremities who develop LR are at a very significantly high risk of developing metastatic disease and dying of the tumor. PMID- 16691624 TI - Intracranial anomalies detected by imaging studies in 30 patients with Apert syndrome. PMID- 16691625 TI - Overgrowth with severe developmental delay following IVF/ICSI: A newly recognized syndrome? AB - We report on a child with postnatal overgrowth, noted from 3 months of age, associated with severe developmental delay and refractory seizures. The patient was conceived using in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). The patient's features do not resemble any reported overgrowth syndromes reported so far. We propose that this is a previously unrecognized disorder. The potential role of ICSI/IVF in the pathogenesis of this condition is unknown. No imprinting defect at chromosome 11p15 was identified. PMID- 16691626 TI - A nonsynonymous single-nucleotide polymorphism in the PDZ-Rho guanine nucleotide exchange factor (Ser1416Gly) modulates the risk of lung cancer in Mexican Americans. AB - BACKGROUND: Based on in vitro studies, Rho guanine nucleotide exchange factors (RhoGEFs) are key regulators of mitogenic and transforming pathways. At least 1 family member, PDZ-RhoGEF, also integrates signaling between monomeric Rho G proteins and heterotrimeric G proteins through a so-called regulator of G-protein signaling (RGS) domain. Recently, the authors reported that 3 single-nucleotide polymorphisms (SNPs) in 2 members of the RGS family were associated with significant reductions in the risk of cancer. METHODS: For the current report, the authors studied the risk of lung cancer associated with a nonsynonymous SNP (rs868188; Ser1416Gly) in PDZ-RhoGEF in a large lung cancer case-control study of 2260 Caucasians and 369 Mexican Americans. RESULTS: Compared with individuals who had the wild-type genotype (AA), Mexican Americans with the variant genotypes (AG and GG) had a significantly reduced risk for lung cancer (odds ratio [OR], 0.57; 95% confidence interval [95%CI], 0.34-0.94). The protective effect appeared to be more evident in younger individuals (OR, 0.42; 95%CI, 0.20-0.91), men (OR, 0.36; 95%CI, 0.18-0.71), and ever smokers (OR, 0.50; 95%CI, 0.29-0.88). A joint effect was observed between Ser1416Gly and polymorphisms in 2 cell-cycle control genes: p53 (intron 3) and cyclin D1 (CCND1). Tallying the variant alleles of the 4 RGS gene SNPs, a gene-dosage effect was apparent. Compared with individuals who had < 3 variant alleles, patients with > or = 3 variant alleles had a 51% reduction in lung cancer risk (OR, 0.49; 95%CI, 0.28-0.88). CONCLUSIONS: To the authors' knowledge, this is the first epidemiological study to link PDZ-RhoGEF polymorphisms with cancer risk. The results suggest that there are interactions between RGS2, RGS6, and PDZ-RhoGEF and validate this family of proteins as key regulators of tumorigenesis. PMID- 16691627 TI - A newly recognized, likely autosomal recessive syndrome comprising agammaglobulinemia, microcephaly, craniosynostosis, severe dermatitis, and other features. AB - We present a novel, likely autosomal recessive, multi-system disorder seen in three siblings, two males and one female, born to nonconsanguineous parents. The disease manifests as agammaglobulinemia with marked microcephaly, significant developmental delay, craniosynostosis, a severe dermatitis, cleft palate, narrowing of the choanae, and blepharophimosis. The constellation of clinical signs seen in this family likely represents a new and recognizable form of agammaglobulinemia due to a defect in early B-cell maturation. PMID- 16691628 TI - Antioxidant potentials of Iranian Carica papaya juice in vitro and in vivo are comparable to alpha-tocopherol. AB - Carica papaya is widely cultivated throughout the world and is used as a food and also as a traditional medicine, particularly as an antiseptic and contraceptive. The present study was designed to explore the toxicological and antioxidant potential of dried C. papaya juice in vitro and in vivo. The oral LD(50) of the juice of C. papaya was determined, and the antioxidant potentials determined by DPPH and FRAP tests. In vivo examination was performed after oral administration of dried papaya juice to rats for 2 weeks at doses of 100, 200 and 400 mg/kg. Blood TBARS and FRAP assays were used to determine the potential of the juice to act against oxidative stress. The acute toxicity test (LD(50)) demonstrated that papaya juice is not lethal up to a dose of 1500 mg/kg after oral administration and thus is considered nontoxic. In treated groups, no sign of toxicity was observed. In vitro evaluation of the antioxidant effects of papaya showed that the highest antioxidant activity (80%) was observed with a concentration of 17.6 mg/mL. Blood lipid peroxidation levels decreased significantly after administration of all doses of papaya juice (100, 200, 400 mg/kg/day) to 35.5%, 39.5% and 40.86% of the control, respectively, compared with a value of 28.8% for vitamin E. The blood total antioxidant power was increased significantly by all doses of papaya juice (100, 200, 400 mg/kg/day) to 11.11%, 23.58% and 23.14% of the control, respectively. The value for vitamin E was 18.44%. This preliminary study indicates the safety and antioxidative stress potential of the juice of C. papaya, which was found to be comparable to the standard antioxidant compound alpha-tocopherol. PMID- 16691629 TI - A novel furanocoumarin from Feroniella lucida exerts protective effect against lipid peroxidation. AB - The protective effect of Feroniella lucida against lipid peroxidation was investigated. Bioassay-guided separation using rat brain as a model led to the isolation of a new antioxidant furanocoumarin named feroniellamin (6) along with five known coumarins. The structure of 6 was characterized by interpretation of spectroscopic data. Feroniellamin inhibited lipid peroxidation with an IC(50) value of 52 microM. PMID- 16691630 TI - Estrogenic activity of standardized extract of Angelica sinensis. AB - Since ancient times, extracts of plants have been used for women's health to prevent menopausal symptoms. The symptoms of menopause have been attributed to a reduction in the amount of estrogen produced by the ovaries. In this study the estrogenic activity of a commercial standardized extract of the roots of Angelica sinensis, used to relieve climacteric symptoms was evaluated using in vivo tests such as the degree of cornification of vaginal epithelium, uterotrophic assays and serum LH concentration in ovariectomized rats. Furthermore, the effects on the estrous cycle in rat were investigated. The results obtained have shown that the administration of a standardized ethanol extract in ovariectomized rats exhibited a stimulation of the uterine histoarchitecture, a significant cornification in the vaginal epithelium and a reduction of serum LH concentration showing the estrogenic nature of the extract. Furthermore, the administration of the extract in intact female rats provoked a significant modification of the vaginal smear in 67% of treated rats. The estrous cycle thus modified was characterized by a prolonged estrus stage with a temporary reduction of the regular cyclicity. PMID- 16691631 TI - Hypolipidemic activity of aqueous extract of Withania coagulans Dunal in albino rats. AB - Administration of an aqueous extract of fruits of Withania coagulans (1 g/kg; p.o.) to high fat diet induced hyperlipidemic rats for 7 weeks, significantly reduced elevated serum cholesterol, triglycerides and lipoprotein levels. This drug also showed hypolipidemic activity in triton induced hypercholesterolemia. The histopathological examination of liver tissues of treated hyperlipidemic rats showed comparatively lesser degenerative changes compared with hyperlipidemic controls. The hypolipidemic effect of W. coagulans fruits was found to be comparable to that of an Ayurvedic product containing Commiphora mukkul. PMID- 16691632 TI - Protective effect of BR-16A, a polyherbal preparation against social isolation stress: possible GABAergic mechanism. AB - The antistress effects of BR-16A, a polyherbal preparation and its interaction with GABAergic modulators against social isolation-induced stress were investigated in the present study. Isolation stress was induced by keeping the mice (Laca strain) individually in each cage for 3 weeks and various drug treatments were given for a period of 5 days before the start of the experiments. The various behavioural parameters examined included pentobarbitone-induced sleep (sleep latency and duration), analgesia (tail-ssick test) and locomotor activity, respectively. BR-16A (100 mg/kg and 200 mg/kg) treatment for 5 days significantly reversed the social isolation stress-induced prolongation of onset and decrease in pentobarbitone-induced sleep, increased total motor activity and stress induced antinociception. When diazepam (0.5 mg/kg), a benzodiazepine agonist, was co-administered with BR-16A (100 mg/kg), it significantly potentiated the reversal of pentobarbitone-induced shortening of sleep time effects; increased locomotor activity and stress induced antinociceptive effects. However, the sleep latency was not decreased significantly. Further, ssumazenil (2 mg/kg), a benzodiazepine receptor antagonist and FG 7142 (10 mg/kg), an inverse agonist, when co-administered with BR-16A (100 mg/kg), showed no significant reversal on pentobarbitone-induced hypnosis, locomotor activity and social isolation-induced antinociception compared with their effects per se. The present study demonstrated the antistress effects of BR-16A preparation against social isolation-induced stress. The present study also suggests that the GABAergic system may be involved in its antistress effect. PMID- 16691633 TI - Umbilical vein constriction at the umbilical ring: a longitudinal study. AB - OBJECTIVE: It has been suggested that constriction of the umbilical vein (UV) at the umbilical ring has hemodynamic effects. We aimed to determine the occurrence and extent of such constriction in serial observations. METHODS: This was a prospective longitudinal study of UV velocities at the umbilicus measured at approximately 4-week intervals between 19 and 42 weeks' gestation in 129 low-risk singleton pregnancies. Each participant was examined three to five times. Multilevel modeling was used to construct the reference ranges and to test associations between variables. RESULTS: Gestational age-specific reference percentiles of UV velocities at the umbilicus were established based on 469 observations. Fetuses were able to alter the UV velocities considerably during the second half of pregnancy, signifying a varying degree of UV constriction. Of a total of 129 fetuses, 56 (43.4%) never had high UV blood velocity (i.e. > 46 cm/s, the highest quartile), 42 (32.6%) fetuses had high UV blood velocity on one occasion and 31 (24.0%) fetuses on two or more occasions. In 36 (27.9%) fetuses the UV velocity at the umbilical ring was > 300% of the mean gestational age specific reference value at the intra-abdominal section on at least one occasion. Constriction of the UV at the umbilical ring did not affect the pulsatility of the umbilical artery, and was not associated with adverse perinatal outcome in this study. CONCLUSIONS: Low-risk fetuses may well constrict the UV at the abdominal wall with velocities extending over wide ranges on one or more occasions during the second half of pregnancy. Rather than being a risk for complications, the constriction seems to be part of physiological development and possibly a regulatory mechanism. PMID- 16691634 TI - Birt-Hogg-Dube gene mutations in human endometrial carcinomas with microsatellite instability. AB - Birt-Hogg-Dube (BHD) syndrome is a rare form of autosomal dominantly inherited genodermatosis characterized by benign hamartomatous skin lesions named fibrofolliculomas, and an increased risk for developing pulmonary cyst/pneumothorax and various forms of renal cell carcinoma. Many of the patients harbour insertion/deletion mutations in the hypermutable poly(C)8 tract in exon 11 of the BHD gene. This mutational hot spot is also reported to be a target of mutation in microsatellite instability (MSI) sporadic colorectal cancer. To test the hypothesis that the BHD gene is also a mutational target in sporadic endometrial carcinoma with microsatellite instability, 139 cases of sporadic endometrial carcinoma were screened for MSI status, and mutations of the poly(C)8 tract in exon 11 as well as other coding exons of the BHD gene. The poly(G)8 tract of the BAX gene, the poly(C)8 tract of MSH6, and methylation status of hMLH1 were also assessed. Thirty-nine of 139 cases (28%) showed MSI. Mutations in the poly(C)8 tract of BHD were detected in five of the 39 MSI cases (12.8%). Of these, one showed additional mutation in exon 4, possibly satisfying the two-hit hypothesis of tumour suppressor genes. BAX gene mutation was detected in ten of the 39 MSI cases (25.6%). Four tumours showed both BAX and BHD mutations, and a significant positive association was found between mutations of the two genes. No association was found between BHD status and MSH6 mutation or hMLH1 methylation. When multiple foci were microdissected and individually screened for mutation, BHD mutations were shown to have been acquired during tumour progression, after mutation of the BAX gene, in three of five cases. Taken together, these findings show that the BHD gene is a target gene in MSI endometrial carcinoma. However, its mutational frequency is lower than that of BAX, and BHD mutation tends to occur during neoplastic progression after the acquisition of mutations in another MSI target gene, BAX. PMID- 16691635 TI - Proliferation of endothelial cell on polytetrafluoroethylene vascular graft materials carried VEGF gene plasmid. AB - OBJECTIVE: To investigate whether vascular endothelial growth factor (VEGF) gene plasmid carried by polytetrafluoroethylene (PTFE) vascular graft materials could transfect endothelial cells (ECs) and promote their growth. METHODS: PTFE vascular graft materials carried with pCDI-hVEGF(121), pCDI or pEGFP were incubated in Tris-buffer solution and the values of optical density of 260 nm at different time were plotted, then the DNA controlled release curve was made. ECs derived from human umbilical vein were seeded on the pCDI-hVEGF(121)/pCDI/pEGFP PTFE materials or tissue culture plates, ECs numbers were counted and VEGF protein concentrations at different time were measured by enzyme-linked immunoadsorbent assay method. Green fluorescent protein (GFP) expression in ECs on pEGFP-PTFE materials was examined with fluorescence microscopy. RESULTS: The controlled release curve showed that the gene released from PTFE materials was rapid within 8 h, then slowed down and that the gene released continuously even after 72 h. At 24, 72 and 120 h, ECs number and proliferation rate of pCDI hVEGF(121)-PTFE materials were higher than those of pCDI or pEGFP-PTFE materials (P<0.05). VEGF protein concentration of pCDI-hVEGF(121)-PTFE materials was higher than that of pCDI or pEGFP-PTFE materials at 6, 24, 72 and 120 h (P<0.01). GFP expression in ECs on the pEGFP-PTFE materials could be detected by fluorescence microscopy. CONCLUSION: PTFE graft can be used as a carrier of VEGF gene plasmid, VEGF gene carried by PTFE can transfect ECs and promote ECs growth. PMID- 16691636 TI - Guava extract (Psidium guajava) alters the labelling of blood constituents with technetium-99m. AB - Psidium guajava (guava) leaf is a phytotherapic used in folk medicine to treat gastrointestinal and respiratory disturbances and is used as anti-inflammatory medicine. In nuclear medicine, blood constituents (BC) are labelled with technetium-99m ((99m)Tc) and used to image procedures. However, data have demonstrated that synthetic or natural drugs could modify the labelling of BC with (99m)Tc. The aim of this work was to evaluate the effects of aqueous extract of guava leaves on the labelling of BC with (99m)Tc. Blood samples of Wistar rats were incubated with different concentrations of guava extract and labelled with (99m)Tc after the percentage of incorporated radioactivity (%ATI) in BC was determined. The results suggest that aqueous guava extract could present antioxidant action and/or alters the membrane structures involved in ion transport into cells, thus decreasing the radiolabelling of BC with (99m)Tc. The data showed significant (P<0.05) alteration of ATI in BC from blood incubated with guava extract. PMID- 16691637 TI - Co-expression of sCD40LIg and CTLA4Ig mediated by adenovirus prolonged mouse skin allograft survival. AB - OBJECTIVE: To investigate the role of simultaneous blockade of CD40/CD40L and B7/CD28 pathways in the immune tolerance via co-expression of sCD40LIg and CTLA4Ig mediated by replication-defective adenovirus. METHODS: Ad-sCD40LIg IRES(2)-CTLA4Ig, replication-defective adenovirus co-expressing sCD40LIg and CTLA4Ig, was constructed and identified. The co-expression of sCD40LIg and CTLA4Ig was evaluated with confocal laser scanning microscope and Western blotting. Skin transplantations of C57BL/6 to BALB/c mice were performed. PBS, Ad Shuttle-CMV and Ad-sCD40LIg-IRES(2)-CTLA4Ig were administered. Skin graft survival was monitored and the mRNA expression of both genes was evaluated in the skin allografts. RESULTS: Ad-sCD40LIg-IRES(2)-CTLA4Ig was constructed successfully and identified. The co-expression of sCD40LIg and CTLA4Ig was identified with confocal laser scanning microscopy and Western blotting. Compared to the skin graft mean survival time (MST) of non-treated group ((5.75+/-0.71) d) or Ad-Shuttle-CMV-treated group ((5.50+/-0.53) d), the skin graft MST was dramatically prolonged in the Ad-sCD40LIg-IRES(2)-CTLA4Ig-treated group ((16.38+/ 1.19) d, P<0.001). The mRNA expression of both genes was detected. CONCLUSION: Ad sCD40LIg-IRES(2)-CTLA4Ig, a replication-defective adenovirus carrying genes encoding sCD40LIg and CTLA4Ig, was constructed. Simultaneous blockade of CD40/CD40L and B7/CD28 costimulatory pathway mediated by replication-defective adenovirus significantly prolonged skin allograft survival in mice. PMID- 16691638 TI - Quantitative real-time RT-PCR detection for CEA, CK20 and CK19 mRNA in peripheral blood of colorectal cancer patients. AB - This study is aimed at establishing a sensitive approach to detect disseminated tumor cells in peripheral blood and evaluate its clinical significance. A total of 198 blood samples including 168 from colorectal carcinoma (CRC) patients and 30 from healthy volunteers were examined by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) to evaluate the expression of carcinoembryonic antigen (CEA), cytokeratin 20 (CK20) and cytokeratin 19 (CK19) mRNA. CEA mRNA was detected in 35.8% of patients and 3.3% of controls, CK20 mRNA in 28.3% of patients and 6.7% of controls, and CK19 mRNA in 41.9% of patients and 3.3% of controls. CEA and CK20 mRNA positive ratio increased with the advancing Dukes stages, but there was no significant difference in positive ratio between any two stages (P>0.05). Also, relatively high positive ratio of CEA, CK20 and CK19 mRNA expression was observed in some CRC patients with earlier Dukes stages. A higher positive ratio was obtained when two or three detection markers were combined compared to a single marker. Our study indicates that quantitative real time RT-PCR detection for CEA, CK20 and CK19 mRNA in peripheral blood is a valuable tool for monitoring early stage dissemination of CRC cells in blood circulation. PMID- 16691639 TI - Binding interactions of pefloxacin mesylate with bovine lactoferrin and human serum albumin. AB - The binding of pefloxacin mesylate (PFLX) to bovine lactoferrin (BLf) and human serum albumin (HSA) in dilute aqueous solution was studied using fluorescence spectra and absorbance spectra. The binding constant K and the binding sites n were obtained by fluorescence quenching method. The binding distance r and energy transfer efficiency E between pefloxacin mesylate and bovine lactoferrin as well as human serum albumin were also obtained according to the mechanism of Forster type dipole-dipole nonradiative energy-transfer. The effects of pefloxacin mesylate on the conformations of bovine lactoferrin and human serum albumin were also analyzed using synchronous fluorescence spectroscopy. PMID- 16691640 TI - Study of cellulolytic soil fungi and two nova species and new medium. AB - This study is aimed at identifying and determining the percentage of occurrence frequency of cellulose decomposing soil fungi. The soil samples were inoculated into culture plates prepared in Sabouraud medium under sterilized conditions and incubated at 30 degrees C for 4 to 7 d. The identified fungal species were incubated in self-designed cellulose medium for testing their cellulolytic ability. Forty-two species, including 2 nova species, representing sixteen genera showed growth and sporulation in the cellulose medium. Most of the isolated species were from genus Aspergillus and Penicillium. Aspergillus niger and Mucor hiemalis showed highest occurrence frequency (45% and 36% respectively), as these species were collected from about 80% of soil samples. Being agar free and cheaper, the new fungal medium designed showed results equivalent to Sabouraud medium. PMID- 16691641 TI - Ciona intestinalis as an emerging model organism: its regeneration under controlled conditions and methodology for egg dechorionation. AB - The ascidian Ciona intestinalis is a model organism of developmental and evolutionary biology and may provide crucial clues concerning two fundamental matters, namely, how chordates originated from the putative deuterostome ancestor and how advanced chordates originated from the simplest chordates. In this paper, a whole-life-span culture of C. intestinalis was conducted. Fed with the diet combination of dry Spirulina, egg yolk, Dicrateria sp., edible yeast and weaning diet for shrimp, C. intestinalis grew up to average 59 mm and matured after 60 d cultivation. This culture process could be repeated using the artificially cultured mature ascidians as material. When the fertilized eggs were maintained under 10, 15, 20, 25 degrees C, they hatched within 30 h, 22 h, 16 h and 12 h 50 min respectively experiencing cleavage, blastulation, gastrulation, neurulation, tailbud stage and tadpole stage. The tadpole larvae were characterized as typical but simplified chordates because of their dorsal nerve cord, notochord and primordial brain. After 8 - 24 h freely swimming, the tadpole larvae settled on the substrates and metamorphosized within 1- 2 d into filter feeding sessile juvenile ascidians. In addition, unfertilized eggs were successfully dechorionated in filtered seawater containing 1% Tripsin, 0.25% EDTA at pH of 10.5 within 40 min. After fertilization, the dechorionated eggs developed well and hatched at normal hatching rate. In conclusion, this paper presented feasible methodology for rearing the tadpole larvae of C. intestinalis into sexual maturity under controlled conditions and detailed observations on the embryogenesis of the laboratory cultured ascidians, which will facilitate developmental and genetic research using this model system. PMID- 16691642 TI - Application of near-infrared spectroscopy to predict sweetpotato starch thermal properties and noodle quality. AB - Sweetpotato starch thermal properties and its noodle quality were analyzed using a rapid predictive method based on near-infrared spectroscopy (NIRS). This method was established based on a total of 93 sweetpotato genotypes with diverse genetic background. Starch samples were scanned by NIRS and analyzed for quality properties by reference methods. Results of statistical modelling indicated that NIRS was reasonably accurate in predicting gelatinization onset temperature (T(o)) (standard error of prediction SEP=2.014 degrees C, coefficient of determination RSQ=0.85), gelatinization peak temperature (T(p)) (SEP=1.371 degrees C, RSQ=0.89), gelatinization temperature range (T(r)) (SEP=2.234 degrees C, RSQ=0.86), and cooling resistance (CR) (SEP=0.528, RSQ=0.89). Gelatinization completion temperature (T(c)), enthalpy of gelatinization (DeltaH), cooling loss (CL) and swelling degree (SWD), were modelled less well with RSQ between 0.63 and 0.84. The present results suggested that the NIRS based method was sufficiently accurate and practical for routine analysis of sweetpotato starch and its noodle quality. PMID- 16691643 TI - Optimization of elastolysis conditions and elastolytic kinetic analysis with elastase from Bacillus licheniformis ZJUEL31410. AB - The solubilization of elastin by Bacillus licheniformis elastase cannot be analyzed by conventional kinetic methods because the biologically relevant substrate is insoluble and the concentration of enzyme-substrate complex has no physical meaning. In this paper we report the optimization of elastolysis conditions and analysis of elastolytic kinetics. Our results indicated that the hydrolyzing temperature and time are very important factors affecting elastolysis rate. The optimized conditions using central composite design were as follows: elastolysis temperature 50 degrees C, elastase concentration 1 x 10(4) U/ml, elastin 80 mg, elastolytic time 4 h. Investigation of the effects of substrate content, elastase concentration and pH was also revealed that low or high elastin content inhibits the elastolysis process. Increasing elastase improves elastin degradation, but high elastase may change the kinetics characterization. Alkaline environment can decrease elastin degradation rate and pH may affect elastolysis by changing elastase reaction pH. To further elucidate the elastolysis process, the logistic model was used to elastolysis kinetics study showing clearly that the logistic model can reasonably explain the elastolysis process, especially under lower elastase concentration. However, there is still need for more investigations with the aid of other methods, such as biochemical and molecular methods. PMID- 16691644 TI - Studies on vibration characteristics of a pear using finite element method. AB - The variation of the vibration characteristics of a Huanghua pear was investigated using finite element simulations. A new image processing technique was used to obtain the unsymmetrical and un-spherical geometrical model of a pear. The vibration characteristics of this type of pear with the correlation of its behavior with geometrical configurations and material characteristics were investigated using numerical modal analysis. The results showed that the eigenfrequency increased with the increasing pear Young's modulus, while decreased with increasing pear density, and decreased with increasing pear volume. The results of this study provided foundation for further investigations of the physical characteristics of fruits and vegetables by using finite element simulations. PMID- 16691645 TI - Evaluation of a kinetic uricase method for serum uric acid assay by predicting background absorbance of uricase reaction solution with an integrated method. AB - A patented kinetic uricase method was evaluated for serum uric acid assay. Initial absorbance of the reaction mixture before uricase action (A(0)) was obtained by correcting the absorbance at 293 nm measured before the addition of uricase solution, and background absorbance (A(b)) was predicted by an integrated method. Uric acid concentration in reaction solution was calculated from A, the difference between A(0) and A(b), using the absorptivity preset for uric acid. This kinetic uricase method exhibited CV<4.3% and recovery of 100%. Lipids, bilirubin, hemoglobin, ascorbic acid, reduced glutathione and xanthine <0.32 mmol/L in serum had no significant effects. A linearly responded to 1.2 to 37.5 micromol/L uric acid in reaction solution containing 15 microl serum. The slope of linear response was consistent with the absorptivity preset for uric acid while the intercept was consistent with that for serum alone. Uric acid concentrations in clinic sera by different uricase methods positively correlated to each other. By Bland-Altman analysis, this kinetic uricase method accorded with that by quantifying the total change of UV absorbance on the completion of uricase reaction. These results demonstrated that this kinetic uricase method is reliable for serum uric acid assay with enhanced resistance to both xanthine and other common errors, wider range of linear response and much lower cost. PMID- 16691646 TI - Statement of Cases of Gonorrhoeal and Purulent Ophthalmia treated in the Desmarres (U. S. Army) Eye and Ear Hospital, Chicago, Illinois, with Special Report of Treatment Employed. PMID- 16691647 TI - Report on the Progress of Ophthalmology during the past year. PMID- 16691648 TI - SPECIALTIES IN MEDICINE. PMID- 16691649 TI - Case of Sudden Monocular Amaurosis. PMID- 16691650 TI - Fracture of the Lens of one Eye and of the Anterior Capsules of both Eyes from Death by Violent Hanging. PMID- 16691651 TI - Extracts from Observations upon Anaesthesia of the Cornea and Radiating Fibres of the Iris occurring without Intra-Ocular Tension. PMID- 16691653 TI - Refraction of a Small Oblique Pencil of Diverging Rays by a Glass Plate with plane parallel Surfaces. PMID- 16691652 TI - Sudden Monocular Amaurosis presenting Unusual Difficulties in Diagnosis. PMID- 16691654 TI - On the Necessity of Employing Greater Accuracy in Ascertaining and Expressing the Degree of Acuteness of Vision. PMID- 16691655 TI - A Method of Operating for Divergent Squint. PMID- 16691656 TI - Amblyopia produced by Osmic Acid. PMID- 16691657 TI - Cases of Sub-Retinal Effusion-Puncture. PMID- 16691658 TI - Anatomy and Physiology of the Ciliary Muscle in Man. PMID- 16691660 TI - Vascular Tumor of Iris, probably Sarcomatous, with Colored Plate. PMID- 16691659 TI - Suture of the Flap after Extraction of Cataract. PMID- 16691661 TI - Remarks on the use of Leaden Styles in the Treatment of Lachrymal Obstructions, with Description of a New Plan for Facilitating their Introduction. PMID- 16691663 TI - Test Types and the Power of Perception. PMID- 16691662 TI - Relative Accommodation in Strabismus and Insufficiency of the Recti Muscles. PMID- 16691664 TI - Monocular Glaucoma supervening on Binocular Retinitis Hemorrhagica. PMID- 16691665 TI - A Modification of the Ophthalmoscope. PMID- 16691666 TI - A Modification of the Ophthalmoscope. PMID- 16691667 TI - An Improved Eye Speculum. PMID- 16691668 TI - A New Form of Cataract Knife. PMID- 16691669 TI - A New Method of Applying Pressure to the Eye. PMID- 16691671 TI - A Question in reference to Photographing the Interior of the Human Eye. PMID- 16691670 TI - A Method of Dressing Eyes after Cataract Extraction, and other Ophthalmic Operations requiring Rest by Exclusion of Light. PMID- 16691672 TI - Apparent Movement of Muscae. PMID- 16691673 TI - Fracture of the Crystalline Lens in Persons Executed by Hanging. PMID- 16691674 TI - A Case of Simple Synchisis. PMID- 16691675 TI - The Inaccuracy introduced into the Determination of Visual Acuteness by disregarding the Magnifying or Diminishing Power of Spectacles. PMID- 16691676 TI - Three Cases of Herpes Zoster Frontalis seu Ophthalmicus. PMID- 16691677 TI - Cases of Foreign Bodies in the Vitreous Humor. PMID- 16691678 TI - Foreign Body in the Eye diagnosticated by Limitation of the Visual Field. PMID- 16691679 TI - Some Curious Phenomena resulting from Reflex Nervous Action in consequence of Traumatic Lesion of the Eye. PMID- 16691680 TI - Quinine as a Local Remedy in Certain Forms of Conjunctival and Corneal Disease. PMID- 16691681 TI - The Channel by which, in Cases of Neuroretinitis, the Exudation proceeds from the Brain into the Eye. PMID- 16691682 TI - Demonstration of Ophthalmoscopic Drawings. PMID- 16691683 TI - The Light Streak seen in the Centre of the Retinal Vessels with the Ophthalmoscope. PMID- 16691684 TI - Ectropium, Exophthalmus, Extirpation, Plastic Operation. PMID- 16691686 TI - Remarks on some Practical Points concerning Cataract Extraction. PMID- 16691685 TI - On the Treatment of Strabismus Convergens through the Accommodation. PMID- 16691687 TI - A Contribution to the Subject of Strabismus. PMID- 16691689 TI - A Case of Cyst of the Iris. PMID- 16691688 TI - Cyst of the Iris cured by Operation, Zinn's Membrane forming its Anterior Wall. PMID- 16691690 TI - Cyst of the Iris. PMID- 16691692 TI - Report on the Progress of Ophthalmology. PMID- 16691691 TI - Results of Thirteen Passavant Operations for breaking up Attachments of the Iris to the Capsule of the Lens. PMID- 16691693 TI - A Case of Supposed Glioma. Occurring in the Clinic of Dr. , at the New York Eye and Ear Infirmary: Erskine Mason. PMID- 16691694 TI - Rough Notes of a Case of Glio-Sarcoma. PMID- 16691695 TI - Tumors of the Retina. PMID- 16691696 TI - A Case of Retinal Glioma, operated on at a very early period, and showing some New and Peculiar Anatomical Conditions. PMID- 16691697 TI - Apparent Form of Inverted Ophthalmoscopic Image of Optic Disk in Astigmatism. PMID- 16691698 TI - A Peculiar Case of Conjunctival and Corneal Inflammation. PMID- 16691699 TI - Canthoplasty in the Treatment of Diphtheritic Conjunctivitis. PMID- 16691700 TI - An Additional Method to Determine the Degree of Ametropia. PMID- 16691701 TI - An Additional Method for the Determination of Astigmatism. PMID- 16691702 TI - On Astigmatism as an Active Cause of Myopia. PMID- 16691703 TI - An Astigmatic Glass for Cataract Patients, with some Remarks on the Statistics of Vision in Cataract Operations. PMID- 16691704 TI - An Apparatus for testing the Perception of Color. PMID- 16691705 TI - Modification of Jaeger's Ophthalmoscope. PMID- 16691707 TI - Detachment of Retina with Laceration at Macula. PMID- 16691706 TI - Gunshot-wound of the Brain, followed by Fungus Cerebri: Hemiopsia. PMID- 16691708 TI - Report on the Progress of Ophthalmology. PMID- 16691709 TI - Report of Passavant Operations with and without Ether, and also under Nitrous Oxide. PMID- 16691710 TI - Readjustment of Levator Muscle of Upper Lid. PMID- 16691711 TI - Additional Note upon the Use of Atropine in the Treatment of Incipient Strabismus. PMID- 16691712 TI - Paralysis of Trigeminus followed by Sloughing of Cornea. PMID- 16691713 TI - A Case of Congenital Fissure of the Lids. PMID- 16691714 TI - Congenital Absence of both Eyeballs: Two Cases. PMID- 16691715 TI - An Apparatus for cutting Microscopic Sections of Eyes. PMID- 16691716 TI - General Syphilitic Inflammation of the Eye. PMID- 16691717 TI - Halo round Macula. PMID- 16691718 TI - A Scheme to aid in recording and examining Cases of Asthenopia. PMID- 16691719 TI - On the Tables given by Loring and Knapp to show the Displacement of the Retina in Ametropia. PMID- 16691720 TI - Variety of Forms of Small Pencils of Astigmatic Rays. PMID- 16691721 TI - The Retina an Asymmetrical Surface. PMID- 16691723 TI - On the Cause of Anaesthesia of the Cornea and Radiating Fibres of the Iris, without Intraocular Tension. PMID- 16691722 TI - Tests for the Insufficiency of the Recti Interni Muscles. PMID- 16691725 TI - Practical Observations on Extraction of Cataracts in Adults. PMID- 16691724 TI - Report on the Progress of Ophthalmology for 1867. PMID- 16691726 TI - On a Color Test for Astigmatism. PMID- 16691727 TI - On a New System of Tests for the Detection and Measurement of Astigmatism, with an Analysis of Sixty-four Cases of Refractive Anomalies observed by the Aid of this Method. PMID- 16691728 TI - Observations in Astigmatism. PMID- 16691729 TI - Case of Acquired Astigmatism. PMID- 16691731 TI - On the Use of Styles of Lead in the Treatment of Disease of the Lachrymal Sac. PMID- 16691730 TI - Modification of Probes for the Lachrymal Passages. PMID- 16691732 TI - Mydriatics in Glaucoma: Two cases in which the instillation of atropine was directly followed by an outbreak of acute glaucoma. PMID- 16691733 TI - Case of Encephaloid Tumor springing from the Fundus of the Eye-ball simulating Glaucoma. PMID- 16691734 TI - Report on the Progress of Ophthalmology for 1865 and 1866. PMID- 16691736 TI - Suture of the Wound of Cornea after Flap Extraction-Report of Forty-four Cases. PMID- 16691735 TI - Relative Accommodation. PMID- 16691738 TI - Inclination of the Vertical Meridian of the Eye-ball for the Direction of Vision upward and outward. PMID- 16691737 TI - A Cystitome for the Modified Linear Extraction. PMID- 16691739 TI - A Point in the Physiology of Vision. PMID- 16691740 TI - On a New Series of Test-Letters for determining the Acuteness of Vision. PMID- 16691741 TI - Retinitis in Glycosuria. PMID- 16691742 TI - A Case of Herpes Zoster Ophthalmicus, in a Patient nearly Eighty Years of Age, causing Fatal Prostration. PMID- 16691744 TI - Enucleation of Eye for Sympathetic Ophthalmia. PMID- 16691743 TI - Remarks on the Operation for Cataract by Modified Linear Extraction-Graefe's Method. PMID- 16691745 TI - Remarks upon the Principle of the Thaumatrope. PMID- 16691746 TI - On the Operation of some Kinds of Complicated Cataract. PMID- 16691747 TI - Influence, on the Refraction, of Four Years of College Life. PMID- 16691749 TI - Case of Sarcoma of the Conjunctiva. PMID- 16691748 TI - Dacryocystitis in Nursing Infants. PMID- 16691751 TI - Prolapse of Retina, in Consequence of Hemorrhage following Iridectomy for Glaucoma. PMID- 16691750 TI - Extirpation in Panophthalmitis. PMID- 16691752 TI - Demonstration of Water-Color Drawings of some of the Rarer Forms of Diseases of the Eye. PMID- 16691753 TI - Paresis of Inferior Oblique Muscle. PMID- 16691754 TI - Tumor of the Optic Nerve. PMID- 16691756 TI - The Diffusion Circles of Ametropia. PMID- 16691755 TI - A Case of Sarcoma of the Choroid, Removed at an Early Stage. PMID- 16691757 TI - Ischaemia of the Retinal Vessels of both Eyes, following Facial Erysipelas. PMID- 16691758 TI - A Case of Apparent Atrophy of the Optic Nerve, in which Recovery of Eyesight followed the Use of Hypodermic Injections of Nitrate of Strychnia. PMID- 16691759 TI - Plastic Operations at the Inner Canthus. Report of Three Cases. PMID- 16691760 TI - A Description of a Modification of Forster's Perimeter. PMID- 16691761 TI - Statistics of Extractions of Cataract by the Flap Operation and by v. Graefe's Method. PMID- 16691762 TI - A Case of Intra-orbital Tumor of Rapid Growth, which resulted in Death. PMID- 16691764 TI - The Treatment of Strictures of the Nasal Duct by means of Large Probes. PMID- 16691763 TI - Hyperostosis and Periostosis of the Bones of the Orbit. Illustrated by two cases. PMID- 16691765 TI - Iridectomy in Chronic Iritis. PMID- 16691766 TI - Three Cases of Restoration of the Eyelid by Transplantation of a Flap without a Pedicle. PMID- 16691767 TI - A Successful Case of Skin Flap Transplantation without Pedicle to Lower Lid, after Removal of an Epithelioma. PMID- 16691768 TI - Case of Gangrene of the Lids, with subsequent Restoration of Tissue without Plastic Operation. PMID- 16691769 TI - Case of Hysterical Monocular Blindness, with violent Blepharospasm and Mydriasis All Relieved by Mental Impression. PMID- 16691770 TI - Neuro-Retinitis, with almost Sudden Loss of Sight in Both Eyes. PMID- 16691772 TI - Jequirity Ophthalmia. PMID- 16691771 TI - Hereditary Atrophy of the Optic Nerves. PMID- 16691773 TI - A New Perimeter. PMID- 16691774 TI - A Case of Embolism of the Retinal Artery. PMID- 16691775 TI - A Case Illustrating the Natural History of Cataract. PMID- 16691777 TI - Osteoma of the Orbit. PMID- 16691776 TI - Myxo-Sarcoma of the Orbit. PMID- 16691778 TI - Clinical History of a Case of Sympathetic Ophthalmia. PMID- 16691780 TI - Experiences in Refractive Cases. PMID- 16691779 TI - Two Cases of Orbital Abscess. PMID- 16691781 TI - Cases of Disease of the Frontal Sinus. PMID- 16691782 TI - A Case of Glioma of the Retina (double-congenital). PMID- 16691783 TI - Two Cases of Swelling of the Optic Papilla-Possibly Congenital. PMID- 16691784 TI - Two Cases in which Rahlmann's Hyperbolic Lenses Improved Vision. PMID- 16691785 TI - A Case of Myxoedema, with Atrophy of the Optic Nerves. PMID- 16691787 TI - A Case of Acute Dacryo-Adenitis. PMID- 16691786 TI - A Report of the Examination of the Employees of the Pennsylvania Rail Road as to Color Sense, Acuteness of Vision, and Hearing. PMID- 16691788 TI - Multiple Colobomata of the Iris, or Polycoria Congenitalis. PMID- 16691790 TI - A Case of Hypermetropic Refraction Passing while under Observation into Myopia Symptoms Simulating General Nervous Disease. PMID- 16691789 TI - Removal of Foreign Bodies from the Vitreous by Aid of the Electro-Magnet. PMID- 16691791 TI - Report of Thirty-Six Cases of Simple Extraction of Cataract without Iridectomy. PMID- 16691792 TI - Report on a Series of One Thousand successive Cases of Extraction of Cataract with Iridectomy. PMID- 16691793 TI - A Case of Amaurosis after the Administration of Large Doses of Quinine-Recovery. PMID- 16691795 TI - Passive Motion in the Treatment of Paralysis of the Ocular Muscles. PMID- 16691794 TI - Clinical Contributions to the Study of Ring-Scotoma. PMID- 16691796 TI - A Case of Recurrent Paralysis of the Motor Oculi. PMID- 16691797 TI - Cases of Primary Acute Glaucoma, of Primary Haemorrhagic Glaucoma, and of Secondary Glaucoma, with Remarks and Photo-Micrographs. PMID- 16691798 TI - Sharply Circumscribed Hyperaemia occurring in Connection with Muscular Insufficiency. PMID- 16691799 TI - Glio-Sarcoma of the Retina; Enucleation; no Recurrence after 14(1/2) Years. PMID- 16691800 TI - Pseudo-Glioma from Traumatism-Enucleation. PMID- 16691801 TI - Transmitted Pulsation at the Fundus Oculi. PMID- 16691803 TI - An Operation for Entropium of the Lower Lid. PMID- 16691802 TI - Spontaneous Prolapse of the Lachrymal Gland.-Its Extirpation. PMID- 16691804 TI - The Eye of the Adult Imbecile. PMID- 16691805 TI - The "Hohlschnitt" of Von Jaeger in the Extraction of Cataract. PMID- 16691806 TI - Cilio-Retinal or Aberrant Vessels. PMID- 16691807 TI - Sarcoma of the Eyelid, Simulating a Meibomian Cyst. PMID- 16691808 TI - Hypermetropic Refraction Passing while Under Observation into Myopia. PMID- 16691809 TI - Note on the Increase of Blindness in the United States. PMID- 16691810 TI - The Pathogenesis of Pterygium. PMID- 16691811 TI - A Case of Recurrent Retinal Haemorrhages, followed by the Outgrowth of Numerous Blood-vessels from the Optic Disc into the Vitreous Humor. PMID- 16691812 TI - Embolism of the Central Retinal Artery. PMID- 16691813 TI - A Danger to the Eyes of Iron-Workers from the Use of Old Hammers. PMID- 16691814 TI - A Case of Retinitis Pigmentosa treated Electrically. PMID- 16691816 TI - Description of a Case of Coloboma of the Iris, Lens and Choroid: With a Study of the Visual Fields. PMID- 16691815 TI - Coloboma of the Optic Nerve and Sheath. PMID- 16691817 TI - Photography of the Interior of the Eye. PMID- 16691818 TI - A Case of Congenital, Zonular, Grayish-White Opacity around the Fovea. PMID- 16691819 TI - Detachment of the Retina in both Eyes with Albuminuria of Pregnancy: Replacement of Retina. PMID- 16691820 TI - Double Chorio-Retinitis, with Partial Degeneration of the Optic Nerve, Associated with Curious Lymph Extravasation into the Retina and Vitreous. PMID- 16691821 TI - A Rare Form of Ophthalmia Granulosa associated with Icthyosis. PMID- 16691822 TI - A Modified Form of Ophthalmoscope, with Cylinders. PMID- 16691823 TI - A Modification of the Refraction Ophthalmoscope, with an Attachment for using Cylinder Lenses. PMID- 16691824 TI - Trial Set of Small Lenses, and a Modified Trial-Frame. PMID- 16691825 TI - On a Transient Myopia occurring in connection with Iritis. PMID- 16691826 TI - A Case of Rapid Development of Lenticular Opacity. PMID- 16691827 TI - Appendix: Note to Dr. Roosa's Case of Amaurosis after the Administration of large doses of Quinine. PMID- 16691829 TI - A Case of Intracranial Neoplasm with Localizing Eye Symptoms, etc. PMID- 16691828 TI - Two Cases of Brain Tumor, with interesting Eye Symptoms. PMID- 16691831 TI - Report of the Committee on the Causes and Prevention of Blindness. PMID- 16691830 TI - The Therapeutic Effect of Prisms in Ophthalmic Practice, etc. PMID- 16691832 TI - Legislation for the Prevention of Blindness. PMID- 16691833 TI - Contributions to Ocular Tumors. PMID- 16691834 TI - The Winking Test. PMID- 16691836 TI - Case of a Foreign Body remaining in the Eye about 20 years, etc. PMID- 16691835 TI - New Tests for Binocular Vision. PMID- 16691837 TI - Foreign Bodies in the Orbit. PMID- 16691838 TI - Simple Extraction of Cataract without Iridectomy. PMID- 16691839 TI - The Extraction of Lenses dislocated into the Vitreous Humor. PMID- 16691840 TI - Removal of a large Exostosis of the Orbit with Preservation of the Eye. PMID- 16691841 TI - Operation for Ectropium of the Lower Lid by the Sliding Flap method. PMID- 16691842 TI - A FORM OF XEROSIS. PMID- 16691844 TI - Two Cases of Glaucoma with Unusual Features. PMID- 16691843 TI - Report of a Case of Recurrent Irido-Chorido-Retinitis. PMID- 16691845 TI - Transient Amblyopia with Bitemporal Hemianopsia in a Case of Malarial Cachexia. PMID- 16691846 TI - Complete Paralysis of the Lateral Movements of both Eyes, etc. PMID- 16691847 TI - An Analysis of the Ocular Symptoms found in the Third Stage of General Paralysis of the Insane. PMID- 16691849 TI - Transient Pulsation of Retinal Arteries induced by Application of Homatropine. PMID- 16691848 TI - New Operation for Symblepharon. PMID- 16691850 TI - A Case of Metastatic Carcinoma of the Choroid. PMID- 16691852 TI - Embolism of Upper Branch of Retinal Artery with Normal Vision. PMID- 16691851 TI - Can Hypermetropia be Healthfully Outgrown? PMID- 16691853 TI - Thrombosis of Arteria Centralis Retinae - large Retino-ciliary artery - Central Vision unimpaired. PMID- 16691854 TI - Progressive Hyperopic Astigmatism. PMID- 16691855 TI - An Elementary Discussion of some Cases of Centrical Refraction, etc. PMID- 16691856 TI - Cyst of the Iris, following a Penetrating Wound, etc. PMID- 16691857 TI - Eight Cases of Rudimentary Zonular Double Cataract. PMID- 16691858 TI - Notes on Twenty-one Cases of Cataract. PMID- 16691860 TI - A New Ophthalmoscope. PMID- 16691859 TI - Specimen of Filaria Oculi Humani. PMID- 16691862 TI - Clinical History of a Case of Spindle-celled Sarcoma of the Choroid with a study of the microscopic condition of the Growth. PMID- 16691861 TI - Gouty Retinitis, Chorio-Retinitis, and Neuro-Retinitis. PMID- 16691863 TI - A Case of Sarcoma of the Choroid, with remarks on the early diagnosis of Intra ocular Sarcoma. PMID- 16691865 TI - A Case of Tumor of the Iris, probably tubercular. PMID- 16691864 TI - A Case of Spindle-celled Sarcoma of the Lid. PMID- 16691866 TI - A Case of traumatic dislocation of the Iris under the unbroken conjunctiva - eye damaged but preserved, Typical sympathetic ophthalmia in the other eye. PMID- 16691867 TI - Indirect Massage of the lens for the artificial ripening of Cataract. PMID- 16691868 TI - Retinitis Punctata Albescens. PMID- 16691869 TI - A series of Wools for the ready detection of Color Blindness. PMID- 16691870 TI - Additional note in reference to the case of useful vision maintained by means of a totally dislocated lens heretofore reported to the Society. PMID- 16691871 TI - The relation of the Patellar Tendon-reflex to some of the Ocular Reflexes found in General Paralysis of the Insane. PMID- 16691872 TI - Removal of Steel from the Vitreous by the Electro Magnet. PMID- 16691873 TI - Suppression of the Visual Image. PMID- 16691874 TI - Case of Congenital Cyst of the orbit with Anophthalmos. PMID- 16691875 TI - An Ocular Mask. PMID- 16691876 TI - A permanent magnet for use in removing Foreign Bodies in the Cornea. PMID- 16691877 TI - Can the Loss of the Eyes by Ophthalmia Neonatorum always be prevented. PMID- 16691878 TI - Clinical Notes on some Refraction and Muscular Cases. PMID- 16691879 TI - Cases of Lateral Heterophoria in which the Vertical-diplopia Test proved untrustworthy. PMID- 16691880 TI - On the removal of hard cataract by Suction. PMID- 16691881 TI - Restoration of the Upper Lid by a Pedicle-flap taken from immediately below the Lower Lid. PMID- 16691882 TI - Case of Pulsating Exophthalmos in which both Common Carotid Arteries were ligated. PMID- 16691883 TI - Memorial of Dr. H. W. Williams. PMID- 16691884 TI - The course and prognosis of Orbital Tumors as influenced by surgical operations for their removal. PMID- 16691885 TI - Tumor of Optic Nerve and Retina. PMID- 16691886 TI - Melano-Sarcoma originating in Ciliary Body. PMID- 16691887 TI - Primary Sarcoma of Iris. PMID- 16691888 TI - Primary Sarcoma of Iris. PMID- 16691889 TI - Bone Sarcoma of Supra-Orbital Margin. PMID- 16691890 TI - Two Cases of Cancerous Tumor of Choroid: 1. Carcinoma of the Choroid-2. Sarcoma of the Choroid. PMID- 16691891 TI - Ivory Exostosis of the Orbit. PMID- 16691892 TI - Tuberculosis of Conjunctiva. PMID- 16691893 TI - Tubercle of Iris: Being a Report of a Case of Tubercle of the Iris, with Remarks Addressed to the Inquiry: Does Tubercle Ever Occur Primarily in the Iris? PMID- 16691894 TI - Congenital Growth at inner Canthus. PMID- 16691896 TI - Embolism of Central Artery of Retina. PMID- 16691895 TI - Persistent Pupillary Membrane. PMID- 16691897 TI - Case of Double Choked-Disc. PMID- 16691899 TI - Case of Double Optic Neuritis. PMID- 16691898 TI - Ophthalmic conditions in case of Cerebellar Tumor. PMID- 16691901 TI - Cyst of Optic Disc. PMID- 16691900 TI - Use of Mercury in Traumatic Irido-Choroiditis. PMID- 16691902 TI - Case of Sympathetic Irido-Cylitis. PMID- 16691903 TI - Case of Suppurative Irido-Choroiditis. PMID- 16691904 TI - Double Chorio-Retinitis following lightning flash. PMID- 16691906 TI - Absolute Alcohol as disinfectant for instruments. PMID- 16691905 TI - Sterilization of Instruments for Cataract Extraction. PMID- 16691907 TI - Rupture of Iris. PMID- 16691908 TI - Angioid Streaks in Retina. PMID- 16691909 TI - Examination of eyes from a case of Pernicious Anaemia. PMID- 16691910 TI - Rare and Fatal Disease of Infancy. PMID- 16691911 TI - Rupture of inferior temporal vein of retina. PMID- 16691912 TI - Pupil Reflex in absolute blindness. PMID- 16691914 TI - Blepharoplasty for both lids by single split flap. PMID- 16691913 TI - Bilateral Necrosis of skin of the eyelids. PMID- 16691915 TI - Dermoid Cysts of the Orbit. PMID- 16691916 TI - Diphtheritic Conjunctivitis. PMID- 16691917 TI - Instrument to measure position of axis of cylindrical lens. PMID- 16691918 TI - Reading with defective vision. PMID- 16691919 TI - Note on Keratitis Punctata Superficialis. PMID- 16691921 TI - Location of fragment of steel in eye, by X Rays. PMID- 16691920 TI - Extraction from Vitreous of copper fragment located by X Rays. PMID- 16691922 TI - Hypopion-Keratitis. PMID- 16691923 TI - Management of Glaucoma. PMID- 16691924 TI - A Case of Chronic Glaucoma with some unusual features. PMID- 16691925 TI - An unusual case of Glaucoma. PMID- 16691926 TI - Lenses for Binocular examination by oblique illumination. PMID- 16691927 TI - Anomalies of the epithelial layer of the crystalline lens and anterior polar cataract. PMID- 16691928 TI - Sympathetic ophthalmia caused by glioma-retinae. PMID- 16691929 TI - Case of spontaneous rupture of the eyeball. PMID- 16691930 TI - Description of the reflecting phorometer and a discussion of the possibilities concerning torsion of the eyes. PMID- 16691931 TI - Congenitally inefficient external rectus with binocular vision. PMID- 16691932 TI - Cyst of the vitreous. PMID- 16691934 TI - Case of primary non-pigmented sarcoma of the upper lid. PMID- 16691933 TI - Case of primary tumor of the optic nerve. PMID- 16691935 TI - Case of expulsive intra-ocular hemorrhage after preliminary iridectomy for cataract. PMID- 16691936 TI - Retinitis albuminurica as a factor in the causation of secondary glaucoma. PMID- 16691937 TI - Cases of metastatic panophthalmitis. PMID- 16691939 TI - A communication upon acoin, a new local anesthetic. PMID- 16691938 TI - Multiple rupture of the pupillary border of the iris with subsequent development of myopic astigmatism. PMID- 16691940 TI - More uniform tests for vision, color-sense, and hearing. PMID- 16691941 TI - Double needle intended to facilitate the discission of secondary cataract. PMID- 16691942 TI - A modified perimeter. PMID- 16691943 TI - A method of converting the alternating current for use in connection with the Johnson magnet. PMID- 16691944 TI - Report of three cases of magnetic extraction of steel from the eyeball through the point of entrance. PMID- 16691945 TI - Clinical measurements of pupilary reaction. PMID- 16691947 TI - Modification of the stereoscope. PMID- 16691946 TI - Is there a "hypermetropia acquisita,". PMID- 16691948 TI - The post-operative history of fifty cases of Simple Chronic Glaucoma. PMID- 16691949 TI - Notes of cases of simple Glaucoma treated by resection of the superior ganglion of the cervical sympathetic, with a report of the microscopical examination of the excised ganglia. PMID- 16691950 TI - A case of Endothelioma of the orbit. PMID- 16691951 TI - Melano-sarcoma of choroid and orbit. PMID- 16691952 TI - Congenital orbital cyst with microphthalmos. PMID- 16691953 TI - Case of metastatic carcinoma of one optic nerve with peculiar degenerations of both nerves: Clinical and Pathological Report. PMID- 16691954 TI - Case of double metastatic carcinoma. PMID- 16691955 TI - PATHOLOGICAL REPORT ON DR. JACK'S CASE OF METASTATIC CARCINOMA OF THE CHOROID. PMID- 16691957 TI - A preliminary study of the ocular changes produced by the injection of pure cultures of the bacillus typhosus into the vitreous chambers of rabbits and guinea-pigs. PMID- 16691956 TI - A histologic study of the crystalline lenses of a hanged criminal. PMID- 16691958 TI - The conjunctival flap and the cataract wound. PMID- 16691959 TI - A simple use of the circle of diffusion in the correction of ametropia. PMID- 16691960 TI - Two unusual complications following cataract extraction, (I) Death, after cataract extraction, from diabetes, (2) Delirium tremens following extraction of cataract. PMID- 16691961 TI - Two cases of detachment of the retina, traumatic in origin, treated by drainage incision. PMID- 16691962 TI - A methyl-alcohol debauch and its results. PMID- 16691963 TI - A case of color-blindness in a railroad employee due to tobacco amblyopia, failing to be recognized by any of the wool tests but immediately detected by lamp. PMID- 16691964 TI - A case of cavernous-sinus thrombosis following grippe-autopsy. PMID- 16691965 TI - Treatment of epithelioma of the eyelids by X-rays. PMID- 16691966 TI - The ideal result to be kept in view in the operative treatment of strabismus in children. PMID- 16691967 TI - The extraction of metallic fragments from the vitreous chamber. PMID- 16691968 TI - The value of partial resection of the tarsal cartilage in the operative treatment of congenital ptosis. PMID- 16691969 TI - Acute circumscribed exudative chorio-retinitis. PMID- 16691970 TI - A contribution to the knowledge of ossification occurring in the eye, with clinical report of cases occurring in the service at Lakeside Hospital. PMID- 16691971 TI - Improvement of vision in amblyopia from non-use. PMID- 16691972 TI - A point in the Dioptrics of astigmatic refraction, illustrated by a new model. PMID- 16691973 TI - The desirability of a uniform notation of the meridians of the eyes and those of the visual field. PMID- 16691974 TI - Tumor of the optic nerve with intra-cranial complications. PMID- 16691975 TI - Temporary ligation of the canaliculi as a means of preventing wound infections in operations on the eye. PMID- 16691976 TI - Progress toward greater uniformity in the examination of the vision, color-sense, and hearing of railroad employees. PMID- 16691977 TI - On the confusion in methods of numbering and using prisms. PMID- 16691978 TI - Additional notes on a case of high astigmatism. PMID- 16691979 TI - Permanent left hemianopsia, sequel of puerperal eclampsia. PMID- 16691980 TI - The use of subconjunctival injections of sublimate in cases of orbital cellulitis. PMID- 16691981 TI - On the primary insertions of the ocular muscles. PMID- 16691982 TI - A case of unilateral hemianopsia in which the Wernicke hemianopsic pupillary reaction was present. PMID- 16691983 TI - Suggestions for a uniform nomenclature of the movements and motor anomalies of the eye. PMID- 16691984 TI - A case of acute glaucoma incited by the use of euphthalmine for diagnostic purposes. PMID- 16691985 TI - Vicarious menstruation into the retina, followed by detachment and retinitis striata. PMID- 16691986 TI - Correction of sixteen diopters of astigmatism by galvano-cautery. PMID- 16691987 TI - A case of acute panophthalmitis following discission of the capsule. PMID- 16691988 TI - Skin-grafting in ophthalmic surgery. PMID- 16691989 TI - A case of leuco-sarcoma of the choroid. PMID- 16691990 TI - Tuberculosis of the eye - two cases. PMID- 16691991 TI - Tuberculosis of the conjunctiva. PMID- 16691992 TI - A type of degeneration involving the central zone of the choroid and retina. PMID- 16691993 TI - Are tenotomies for hyperphoria necessarily more uncertain in their results than those for esophoria and exophoria? PMID- 16691994 TI - Interstitial keratitis complicating ophthalmia neonatorum. PMID- 16691995 TI - Argyrosis of conjunctiva and lachrymal sac, following use of protargol. PMID- 16691996 TI - A case of double traumatic optic neuritis, followed by absolute blindness and recovery. PMID- 16691997 TI - A case of traumatic ptosis, operated according to method of Dr. Gruening or Gillet de Grandmont. PMID- 16691998 TI - A case of retinitis pigmentosa. PMID- 16691999 TI - A case of orbital cellulitis, with empyema of the ethmoid and frontal sinuses; death. PMID- 16692000 TI - A case of spontaneous prolapse of both lachrymal glands. PMID- 16692001 TI - Optic atrophy following intestinal hemorrhage. PMID- 16692002 TI - An improved lantern for testing color-perception. PMID- 16692003 TI - Report of the committee on standard test-types and on reading tests. PMID- 16692004 TI - Appendix to report. PMID- 16692005 TI - The present status of subconjunctival injections in ophthalmic therapeutics. PMID- 16692006 TI - Case of pulsating exophthalmos, of traumatic origin; ligation of the common carotid; recovery. PMID- 16692007 TI - Concerning the possible etiological factors in tobacco amblyopia revealed by an analysis of the urine of cases of this character. PMID- 16692008 TI - Hypertrophy and degeneration of the meibomian glands: I. Subsequent History of the Case of Adenoma of the Meibomian Glands, Reported at the Meeting of this Society in 1901. PMID- 16692009 TI - Why not employ intra-capsular irrigation in cataract operations? PMID- 16692010 TI - Memoir of Dr. W. F. Norris. PMID- 16692011 TI - Orbital-osteoma of ethmoidal origin. PMID- 16692012 TI - Diseases of the eye in the white and negro races. PMID- 16692013 TI - Operations upon the eyeball in the presence of an infected conjunctival sac. PMID- 16692014 TI - Concerning certain non-traumatic perforations of the macula lutea. PMID- 16692015 TI - Conjunctivitis nodosa, with histological examination. PMID- 16692016 TI - Intra-ocular tuberculosis with report of two cases. PMID- 16692017 TI - Case of conglomerate tubercle of the choroid. PMID- 16692018 TI - A case of congenital intra-ocular tumor containing epithelium and cartilage. PMID- 16692019 TI - A severe case of uveitis treated with radium. PMID- 16692020 TI - Some observations on the prognosis and treatment of hypopion-ulcer of the cornea. PMID- 16692021 TI - An uncommon congenital anomaly in the vitreous chamber and the inner membranes of both eyes. PMID- 16692022 TI - The bacteriological diagnosis of the diphtheria bacillus especially in conjunctivitis. PMID- 16692023 TI - A case of mind-blindness unique in that the entire mesial surface of both occipital lobes and both optic radiations were preserved. PMID- 16692024 TI - The cortical fusion of some color sensations. PMID- 16692025 TI - A case of ophthalmoplegia-externa bi-lateral. PMID- 16692026 TI - A case of simple glaucoma with some unusual features. PMID- 16692027 TI - Prism exercises, their indications and technique. PMID- 16692028 TI - The pathologic results of dextrocularity and sinistrocularity. PMID- 16692029 TI - The muscle of Horner and its relation to the retraction of the caruncle after tenotomy of the internal rectus. PMID- 16692030 TI - Cystadenoma of the lachrymal gland. PMID- 16692031 TI - Traumatic emphysema of orbit and lids. PMID- 16692032 TI - The act of winking and its photographic measurements. PMID- 16692033 TI - Sympathetic neuro-retinitis and serous uveitis following enucleation with implantation of glass globe, resection of optic nerve - recovery. PMID- 16692034 TI - Angelucci's modification of the technique of the cataract operation - fixation of superior rectus and completion of entire operation without speculum or aid of assistant. PMID- 16692035 TI - A rare tumor arising from the pars ciliaris retinae (terato-neuroma) of a nature hitherto unrecognized, and its relation to the so-called glioma-retinae. PMID- 16692036 TI - The importance of testing the ocular muscle-balance for near as well as for distant vision. PMID- 16692037 TI - Observations of a case of bi-temporal hemianopsia with some unusual changes in the visual fields. PMID- 16692038 TI - Report of two cases of family macular degeneration of the cornea. PMID- 16692039 TI - Symmetrical enlargement of parotid and lachrymal glands - nodular iritis. PMID- 16692040 TI - Excision of the superior cervical ganglion of the sympathetic for simple glaucoma. PMID- 16692041 TI - Report of the Committee on Standards. PMID- 16692042 TI - Improvements in the refractometer for the correction of ametropia described in the Transactions of 1902. PMID- 16692043 TI - Melano-sarcoma of the choroid. PMID- 16692044 TI - Concerning melanoma of the choroid with report of one case of this character and of another exhibiting a pigmented sarcoma of the choroid early in its development. PMID- 16692045 TI - Metastic carcinoma of ciliary body. PMID- 16692046 TI - The Sachs lamp for transillumination of the eye. PMID- 16692047 TI - Improved surgical methods for the successful use of the electromagnet. PMID- 16692048 TI - Magnetic properties of steel alloyed with other metals. PMID- 16692049 TI - On the various methods employed for localizing foreign bodies in the eye by means of the Roentgen rays. PMID- 16692050 TI - What are the so-called reflexes which can properly be referred to eye-strain? PMID- 16692051 TI - Recurrent iritis, a study of nine cases. PMID- 16692052 TI - Juvenile glaucoma. PMID- 16692054 TI - Case of cystic sarcoma of the orbit - extirpation - death. PMID- 16692055 TI - Certain congenital affections of the eye following strong ante-natal impressions made on the mother. PMID- 16692053 TI - Case of acquired cyst of conjunctiva containing an embryonic toothlike structure. PMID- 16692056 TI - Lesion of the choroid due to intestinal inflammation from ptomaine poisoning. PMID- 16692057 TI - The treatment of inoperable cases of malignant disease of the orbit by the X-ray. PMID- 16692058 TI - Vernal conjunctivitis in the negro. PMID- 16692060 TI - Tumors of the conjunctiva and cornea - two cases. PMID- 16692059 TI - Value of the so-called high frequency currents in certain ocular conditions with illustrative cases. PMID- 16692061 TI - A case of homonymous, left-sided inferior tetartanopsia following toxic dose of salicylate of soda. PMID- 16692062 TI - Epithelial corneal cyst. PMID- 16692063 TI - Case of persistent pupillary membrane of rather considerable size. PMID- 16692064 TI - Case of discoloration of the cornea by blood pigment and one of hemorrhage into the cornea. PMID- 16692065 TI - A series of reversible test-types arranged according to the geometrical progression of Drs. Jno. Green. PMID- 16692066 TI - Symptoms presented by the different bacteriological types of acute conjunctivitis. PMID- 16692068 TI - Notes on the clinical determination of the acuteness of vision, including the construction and gradation of optotypes, and on systems of notation. PMID- 16692067 TI - Cyanosis retinae. PMID- 16692069 TI - Retinitis proliferans with development of a system of blood-vessels upon the optic nerve. PMID- 16692070 TI - Case of congenital cyst of the orbit with microphthalmos. PMID- 16692071 TI - Multiple myeloma, with involvement of the orbit. PMID- 16692072 TI - Transient myopia due to trauma. PMID- 16692073 TI - A study of the nativity, sex, age, occupation, and social condition of 3,436 cases of senile cataract operated upon at the Wills Hospital in Philadelphia. PMID- 16692074 TI - Pathological report on Dr. May's case of microphthalmos with upper-lid cyst. PMID- 16692076 TI - Case of invagination cyst of the cornea and of anterior chamber. PMID- 16692075 TI - Concerning siderosis bulbi. PMID- 16692077 TI - Extraction of cataract in the capsule: With Report of ten cases. PMID- 16692078 TI - Silver preparations in conjunctival disease. PMID- 16692079 TI - Extraction of lens in capsule as a method of procedure in cases of immature cataract. PMID- 16692080 TI - Course and treatment of simple cyclitis. PMID- 16692081 TI - Case of secondary hypernephroma of the iris and ciliary body. PMID- 16692082 TI - Case of sarcoma of choroid, with observation on the use of the transilluminator. PMID- 16692083 TI - Ulceration and destruction of cornea by bacillus pyocyaneus. PMID- 16692084 TI - Bacillus pyocyaneus, its virulence in the eye, its longevity and immunity from it. PMID- 16692086 TI - Optic neuritis after injury or disease of the anterior half of the eyeball. PMID- 16692085 TI - An experimental study on the bactericidal power of various silver preparations. PMID- 16692087 TI - Case of pronounced inward and downward squint in near vision with binocular fixation and normal muscle-balance in distant vision. PMID- 16692088 TI - Report of cases of polio-encephalitis superior. PMID- 16692089 TI - An improved electric ophthalmoscope. PMID- 16692090 TI - Plexiform neuroma of orbit. PMID- 16692091 TI - Case of steel in the eyeball which the Haab magnet failed to remove. PMID- 16692092 TI - The modern operation in frontal sinus disease affecting the orbit with two cases. PMID- 16692093 TI - Bacteriological examinations of the conjunctival sac in typhoid fever and in pneumonia: Based upon an examination of 100 cases of Typhoid Fever and 48 cases of Lobar Pneumonia occurring in the wards of the Johns Hopkins Hospital. PMID- 16692094 TI - Case of congenital dislocation of both lenses,-attempted removal by absorption resulting in failure followed by extraction. PMID- 16692095 TI - The immersion treatment with argyrol of the purulent ophthalmias. PMID- 16692096 TI - Operation for removal of nasal epithelioma followed by panophthalmitis. PMID- 16692097 TI - Preliminary note on enucleation of eyeball under local anesthesia. PMID- 16692098 TI - Unilateral rotary nystagmus. PMID- 16692099 TI - A tangent-plane for accurately mapping scotomata and the fields of fixation and single vision, and for indicating the precise position of double images in paralysis. PMID- 16692100 TI - The ocular symptoms in a case of tumor of the pituitary body. PMID- 16692101 TI - Keratitis tuberculosa relieved by tuberculin injections: Report of a Case. PMID- 16692102 TI - Concerning the signs in the retina of persistent high arterial tension and their diagnostic and prognostic import. PMID- 16692103 TI - The value of systematic bacteriologic and microscopic study in the treatment of gonococcal conjunctivitis. PMID- 16692104 TI - Perivasculitis retinae. PMID- 16692105 TI - Vasculitis and perivasculitis. PMID- 16692106 TI - Recurrent conjunctivitis produced by animals. PMID- 16692108 TI - Congenital inequality of the size of cornea in normal eyes. PMID- 16692107 TI - Sarcoma of corneal limbus. PMID- 16692109 TI - Removal of lens in high myopia. PMID- 16692111 TI - Acute inflammatory glaucoma without excavation of the papilla. PMID- 16692110 TI - Post-operative history of sixty cases of simple chronic glaucoma. PMID- 16692112 TI - Proliferative uveitis. PMID- 16692113 TI - Retinitis proliferans in a detached retina. PMID- 16692114 TI - A simple method of representing ocular muscle imbalance by diagram. PMID- 16692115 TI - Unusual form of hereditary congenital cataract in several members of a family. PMID- 16692116 TI - Vascular changes in uveal tract, retina and optic nerve in quinine blindness, and in tobacco-alcohol amblyopia, etc. PMID- 16692117 TI - Affections of the optic nerve during pregnancy. PMID- 16692118 TI - Chronic cyanotic polycythemia. PMID- 16692119 TI - Intra-ocular tuberculosis simulating glioma of retina. PMID- 16692120 TI - Two cases of supposed intra-ocular tuberculosis. PMID- 16692121 TI - Double perforation of the eyeball by iron and steel. PMID- 16692123 TI - Foreign body in fundus - extraction by magnet - complete post-operative aniridia recovery. PMID- 16692122 TI - Double perforation of eyeball by metallic foreign body, removal with preservation of normal vision. PMID- 16692124 TI - Interesting cases of eye-strain. PMID- 16692125 TI - Case of unilateral nystagmus. PMID- 16692126 TI - Position of the ophthalmologist in the treatment of diseases of the accessory sinuses of the nose. PMID- 16692127 TI - Acquired cataract in childhood. PMID- 16692129 TI - Secondary glaucoma in a curious case of keratitis. PMID- 16692128 TI - Pulsating exophthalmus, Iigation of common carotid-death. PMID- 16692130 TI - A new trial frame. PMID- 16692131 TI - Sudden obstruction of the central artery of the retina, being a clinical record of five cases. PMID- 16692132 TI - Embolism of a macular artery and thrombosis of superior and inferior retinal arteries, in a case of embolic softening of the brain. PMID- 16692133 TI - Angioma of the orbit with invasion of the globe along the ciliary nerves. PMID- 16692134 TI - Macular holes. PMID- 16692135 TI - A case exhibiting a symblepharon, corneal opacities and a probably fibro-fatty tumor of the conjunctiva, all of pre-natal origin, in a woman with hereditary cataracts at an early age. PMID- 16692136 TI - Cyanosis retinae. PMID- 16692137 TI - A case of grill-like keratitis. PMID- 16692138 TI - The increasing importance of tuberculosis as a cause of ocular disease, the newer methods of diagnosis and the treatment. PMID- 16692139 TI - Angioma of the choroid. PMID- 16692140 TI - The teleology and interpretation of nystagmus. PMID- 16692141 TI - Two cases of sympathetic ophthalmia. PMID- 16692142 TI - Diffuse interstitial keratitis in acquired syphilis. PMID- 16692143 TI - Recent traumatic erosion of the cornea: Report of a Case due to Lachrymal Stricture. PMID- 16692144 TI - Thyroid extract in keratitis. PMID- 16692145 TI - The treatment of immature cataract. PMID- 16692146 TI - The extraction of cataract with a lance-shaped knife. PMID- 16692147 TI - An attempt to determine the normal range of accommodation at various ages, being a revision of Donder's experiments. PMID- 16692148 TI - The clinical measurement of relative accommodation. PMID- 16692149 TI - Some practical points in the use of prisms. PMID- 16692150 TI - Strabismus from the operative standpoint. PMID- 16692152 TI - Restoration of sight after nearly a month's blindness from glaucoma. PMID- 16692151 TI - A simplification of De Grandmont's operation for ptosis. PMID- 16692153 TI - Bilateral optic neuritis after ethmoiditis. PMID- 16692154 TI - Ocular affections associated with disease of the sinuses contiguous to the orbit. PMID- 16692155 TI - Mixed streptococcus and pneumococcus infections of the orbit and adjacent sinuses. PMID- 16692156 TI - Hemorrhage from the cornea in glaucoma. PMID- 16692157 TI - Adeno-carcinoma of orbit. PMID- 16692158 TI - Blepharo-conjunctivitis caused by diplococcus of Morax-Axenfeld. PMID- 16692159 TI - Description of cases of shrinkage of the eyeball, with remarks on the processes underlying atrophia bulbi in general. PMID- 16692161 TI - The Treatment of the Eye when the Globe is Infected, with the Object of Preventing Panophthalmitis. PMID- 16692160 TI - Report of committee on ophthalmia neonatorum. PMID- 16692162 TI - Third Series of Cases of Injuries from Foreign Bodies Examined by the Roentgen Rays, with Results of Operation. PMID- 16692163 TI - Physical Economics: Comprising a Mathematical Formula for Determining the Earning Ability of a Person which May be used Either to Rate Him, or to Obtain his Economic Value, and to Ascertain Damages to any Part or Parts of his Body from Injury or Disease in a Manner Equitable to all Concerned. PMID- 16692164 TI - Spontaneous Rupture of the Eyeball. Report of a Case. PMID- 16692165 TI - Adenocarcinoma of the Pituitary Body. Report of a Case Under Observation for Four and a Half Years. Pathological Examination. PMID- 16692166 TI - Report of a Case of Sarcoma and Bone Formation in an Atrophic Eye, with Presentation of Specimen. PMID- 16692167 TI - Metastatic Carcinoma of the Orbit, with the Report of a Case. PMID- 16692168 TI - Orbital Fibroma, with Unusual Clinical Manifestations. PMID- 16692169 TI - Symmetrical Lymphomata of the Lacrimal and Salivary Glands (Mikulicz's Disease). PMID- 16692170 TI - A Case of Complete Albinism; Observations on the Changes in the Diameters of the Lens as Seen through the Iris. PMID- 16692172 TI - Hemorrhagic Central Chorioretinitis in Non-myopic Eyes: A Clinical Contribution. PMID- 16692171 TI - A Case of Bilateral Macular Coloboma. PMID- 16692173 TI - Angioid Streaks in the Retina, with the Report of a Case. PMID- 16692174 TI - Concerning a Form of Retinal Disease with Extensive Exudation; Being a Clinical Contribution, with the Report of Two Cases. PMID- 16692175 TI - Neuroretinitis in Chlorosis, with Report of a Case. PMID- 16692176 TI - The Differentiation of the Diphtheria Bacillus from Organisms Morphologically Similar. PMID- 16692177 TI - A Classification of Eye Diseases: With the Outline of a Universal Morbidity List Based on the Nomenclature of Diseases of the Royal College of Physicians, of London. PMID- 16692178 TI - Tropometers and Measurements of Arcs of Rotation. PMID- 16692179 TI - Improved Apparatus for Locating Foreign Bodies in the Eyeball by the Roentgen Rays. PMID- 16692180 TI - Improved Lantern for Testing the Color-Perception. Model of 1909. PMID- 16692181 TI - The Adverse Influence of Diabetes in Certain Operations on the Eye. PMID- 16692183 TI - The Choice of Operation for Iridotomy. PMID- 16692182 TI - Summary of Results Obtained, and Features of Interest, in Two Hundred and Fifteen Consecutive Cataract Extractions. PMID- 16692184 TI - The Post-Operative History of Eighteen Cases of Magnetic Foreign Bodies Removed from the Eye by the Haab or Giant Magnet. PMID- 16692186 TI - Large Piece of Wood Embedded Deeply in Orbit of Child Twenty-five Months. Removed, with Preservation of Vision. PMID- 16692185 TI - The Possible Influence of Racial and Other Characteristics in Accounting for the Success of the Smith-Indian Operation. PMID- 16692187 TI - Conjunctival Flaps in Ophthalmic Surgery. PMID- 16692188 TI - Report of a Case of Leukosarcoma of the Iris Successfully Removed by Iridectomy and Useful Vision Retained. PMID- 16692189 TI - New-Growths of the Lacrimal Gland-A Case. Death: Pathologic Report by Dr. F. H. Verhoeff. PMID- 16692191 TI - Report of a Case of Epibulbar Sarcoma. PMID- 16692190 TI - Congenital Pigmentation of the Cornea. PMID- 16692192 TI - Cylindroma of the Orbit. PMID- 16692193 TI - Tumor of the Adrenal Gland with Metastasis in the Orbit: Pathologic Examination by Dr. H. F. Verhoeff. PMID- 16692194 TI - A Critical Study of the Ocular Asymmetry of the Formosan Savage. PMID- 16692195 TI - Some Impressions of Certain Eye Affections in the Negro as Compared with the White Race. PMID- 16692196 TI - The Difficulty Sometimes met with in the Diagnosis of a Subretinal Mass. PMID- 16692197 TI - Glaucoma from Adhesion of the Lens Capsule to the Cornea. PMID- 16692198 TI - Some Observations of the Ocular Manifestations of Sinus Disease. PMID- 16692199 TI - Implantation of a Metal Ball in Tenon's Capsule (The Frost-Lang Operation) with Report of 48 cases. PMID- 16692200 TI - Report of Two Cases of Chronic Cyanosis with Polycythemia, including Ophthalmoscopic Examination. PMID- 16692201 TI - The Nerve-Head in Wood-Alcohol Amaurosis. PMID- 16692202 TI - Ocular Signs Frequently Associated with Vasomotor Disturbances. PMID- 16692203 TI - A Case of Spasm of the Retinal Arteries. PMID- 16692204 TI - Some Observations on the Use of the Schiotz Tonometer. PMID- 16692205 TI - The Histologic Findings in a Case of Tuberculous Cyclitis, and a Theory as to the Origin of Tuberculous Scleritis and Keratitis. PMID- 16692206 TI - Recurrent Tubercular Choroiditis. PMID- 16692207 TI - Vaccine and Serum Therapy in Ophthalmology. PMID- 16692208 TI - Vaccine and Serum Therapy in Ocular Tuberculosis. PMID- 16692209 TI - Suggestions Regarding the Nature of Visual Sensations. PMID- 16692210 TI - A Window Resection of the Nasal Duct in Cases of Stenosis. PMID- 16692211 TI - Illuminated Box Eye-Testing Chart. PMID- 16692212 TI - A Brief Note Concerning the Tropometer. PMID- 16692213 TI - On the Different Prism Tests, with Presentation of a Series of Prisms Arranged on a Circular Disc. PMID- 16692214 TI - Standardizing of Test-Types and Records of Visual Acuity. PMID- 16692215 TI - Colored Toric Glass for Use in Examining Cases of Ocular Paralysis. PMID- 16692216 TI - Note on the Heredity of Corneal Astigmatism and of Muscular Anomalies. PMID- 16692217 TI - APPENDIX: Report of the Committee on Ophthalmia Neonatorum. PMID- 16692218 TI - The Field of Vision in Tabetic Atrophy of the Optic Disc. PMID- 16692219 TI - A Report of Four Cases of Acute Disseminated Myelitis with Retrobulbar Degeneration of the Optic Nerves. PMID- 16692220 TI - Episcleritis Periodica Fugax. PMID- 16692221 TI - Dystrophia Epithelialis Corneae (Fuchs), with the Report of a Case. PMID- 16692222 TI - The Use of Serum in Sympathetic Ophthalmia. PMID- 16692223 TI - A Case of Gonorrheal Conjunctivitis, with Arthritis Preceding Conjunctival Discharge by Four Days. PMID- 16692224 TI - A Protest Against the Indiscriminate Use of the Organic Compounds of Silver in Ophthalmic Practice. PMID- 16692225 TI - The Operation of Excision of Tarsus and Conjunctiva (Heisrath) in Old Trachoma, Together with Demonstration of a New Instrument, and a Description of a New Method of Performing It. PMID- 16692226 TI - A Case of Ring Sarcoma of the Ciliary Body. PMID- 16692227 TI - Neuroma of the Ciliary Nerves. PMID- 16692228 TI - Smallest Sarcoma of the Choroid. PMID- 16692229 TI - The Pathology of Superficial Punctate Keratitis, with Remarks on Neuropathic Keratitis in General, and on a Hitherto Undescribed Lesion of the Iris. PMID- 16692230 TI - Some Questions Concerning the Method of Development and the Pathology of the Retinitis of Bright's Disease. PMID- 16692231 TI - Probable Cyst of the Retina, with a Demonstration of the Patient. PMID- 16692232 TI - Report of Two Cases of Pulsating Exophthalmos. Relief of one by Ligation of the Common Carotid; Relief of the Other by Ligation of the Common Carotid, Followed after Recurrence by Ligation of the Internal Carotid. PMID- 16692233 TI - Concerning Certain Tumors of the Orbit, and Certain Conditions Simulating Neoplasms of the Orbit, Being a Clinico-pathologic Contribution, with the Report of Six Cases. PMID- 16692234 TI - Subsequent History of a Case of Exostosis of the Orbit Reported to the Society in 1897. PMID- 16692235 TI - Phantom Intraocular Tumor. PMID- 16692237 TI - Intracapsular Extraction of the Cataractous Lens. PMID- 16692236 TI - The Position of the Eye the Element of Safety in the East Indian Extraction in Capsule. PMID- 16692238 TI - The Iris in the Intracapsular Operation for Cataract. PMID- 16692239 TI - Cataract Extraction with the Least Trauma; Excellent Visual Results. PMID- 16692240 TI - The Mechanics of Choked Disc. PMID- 16692241 TI - The Relation of Choked Disc to Intraocular Tension: A Clinical Study of Six Cases. PMID- 16692242 TI - Intestinal Sepsis as an Etiologic Factor in Cardiovascular Disease and Associated Ocular Affections. PMID- 16692243 TI - Cilioretinal and Other Anomalous Retinal Vessels. PMID- 16692244 TI - Exhibition of a Bjerrum Stick. PMID- 16692245 TI - A Double Coloboma (Nasal and Temporal) of the Optic Nerve and Sheath: Exhibition of Drawing. PMID- 16692246 TI - Retinitis Pigmentosa Associated with Albuminuric Neuroretinitis. PMID- 16692247 TI - Embolism of the Right Central Retinal Artery in a Boy Aged Eight Years. PMID- 16692248 TI - Congenital Deviations of the Eyes. PMID- 16692249 TI - Report of Six Cases of Degeneration of the Cornea in the Same Family (Nodular Keratitis). PMID- 16692250 TI - Idiopathic Hematoma of the Orbit. PMID- 16692251 TI - Calcareous Degeneration of the Cornea and Lens Capsule. PMID- 16692253 TI - Polypoidal Formation in the Lacrimal Sac. PMID- 16692252 TI - Fracture of the Skull with Hemorrhage into the Optic Nerve-Sheaths and Retinas: Microscopic Examination of the Eyeballs. PMID- 16692254 TI - A Case of Acute Suppurative Dacryoadenitis. PMID- 16692255 TI - A Case of Bitemporal Hemianopsia with Acromegaly and Other Symptoms Apparently Due to Disease in the Pituitary Region: Diagnosis Confirmed by Autopsy. PMID- 16692256 TI - Amblyopia from Inhalation of Methyl Alcohol. PMID- 16692257 TI - Hereditary Optic Atrophy with x-Ray Findings. PMID- 16692258 TI - A Case of Transient Blindness, Complete in One Eye, Partial in the Other, with Double Optic Neuritis. PMID- 16692259 TI - Flat Sarcoma of the Choroid. PMID- 16692261 TI - A Case of Primary Intradural Tumor of the Optic Nerve. PMID- 16692260 TI - Sarcoma of the Eye: Report of Three Cases. PMID- 16692262 TI - A Case of Plexiform Neurofibroma Involving the Orbit. PMID- 16692263 TI - A Case of Endothelial Sarcoma of the Orbit. PMID- 16692264 TI - A Case of Chloroma. PMID- 16692265 TI - The Treatment of Trachoma with Radium. The Use of Radium-coated Celluloid Plates for This Purpose. PMID- 16692266 TI - Recurrent Retinal Hemorrhages Occurring in the Young, with the Report of a Case. PMID- 16692268 TI - Management of Capsule in Cataract Extraction and Afterward. PMID- 16692267 TI - Some Suggestions to Those Upon Whose Aid the Success of the Ophthalmic Surgeon, in Considerable Measure, Depends. PMID- 16692269 TI - The Relative Value of Recent Improvement of Cataract Extraction. PMID- 16692270 TI - Microscopic Findings in the Eyeball of a Man Who Died from an Untoward Cause Seven Weeks after the Extraction of a Senile Cataract. PMID- 16692271 TI - The Operation of Combined Iridectomy and Sclerectomy for Chronic Glaucoma. PMID- 16692272 TI - A Piece of Glass in the Crystalline Lens, with Description of the Eye Three Years and a Half after the Accident. PMID- 16692273 TI - Degeneration of the Corneas of a Man and His Adult Son. PMID- 16692274 TI - An Extreme Case of Conical Cornea with Cataract. PMID- 16692275 TI - Keratoconus, with Reports of Cases. PMID- 16692277 TI - Ophthalmia Artefacta. PMID- 16692276 TI - Interstitial Keratitis of Luetic Origin. PMID- 16692278 TI - The Prophylaxis of Ocular Birth Infections. PMID- 16692279 TI - An Unusual Case of Staphylococcus Conjunctivitis. PMID- 16692280 TI - A Case of Congenital Apron of the Palpebral Conjunctiva. PMID- 16692281 TI - Report of a Case of Bilateral Congenital Ptosis Relieved by the Motais Operation. PMID- 16692282 TI - Discission of the Crystalline Lens. PMID- 16692283 TI - Two Cases of Simple Chronic Glaucoma Treated by Iridotasis. PMID- 16692284 TI - Fourth Series of Cases of Injuries from Foreign Bodies Examined by the Roentgen Rays, with Results of Operation. PMID- 16692285 TI - A Contribution to the History of the Magnet as Applied to Ophthalmic Surgery. PMID- 16692286 TI - New Cataract Glasses. PMID- 16692287 TI - Electric Operating Lamp. PMID- 16692288 TI - Posterior Ocular Transilluminator. PMID- 16692289 TI - Battery Attachment for Electric Ophthalmoscope. PMID- 16692290 TI - A Second Chapter in the History of a Subretinal Mass (with patient). PMID- 16692291 TI - Two Cases of Sympathetic Ophthalmia. PMID- 16692292 TI - Two Cases of Binocular Coloboma of the Optic Nerve in the Same Family. PMID- 16692293 TI - The Experimental Production of Sclerokeratitis and Chronic Intraocular Tuberculosis. PMID- 16692294 TI - On Some Forms of Retinal Tuberculosis. PMID- 16692295 TI - Two Unusual Orbital Conditions. PMID- 16692296 TI - Report of a Case of Cavernous Sinus Thrombosis Resulting from a Small Abscess in the Skin of the Temple. Vaccine Treatment. PMID- 16692297 TI - Sarcoma of the Choroid not Demonstrable by the Ordinary Transilluminator. PMID- 16692298 TI - Epibulbar Carcinoma; Histologic Examination of the Specimen. PMID- 16692299 TI - Toxic Amblyopia Due to Tobacco Alone. PMID- 16692300 TI - Report of a Case of Paralysis of Divergence. PMID- 16692301 TI - Some Striking Examples of Subnormal Accommodative Power. PMID- 16692303 TI - A Case of Thrombosis of the Retinal Vein, with Remarks. PMID- 16692302 TI - The Eye Examination a Help to Diagnosis of Commencing Arteriosclerosis. PMID- 16692304 TI - One More Word on Test-cards and Type. PMID- 16692305 TI - On the Fatigue of Convergence and Its Varieties. PMID- 16692306 TI - A New Operation for the Removal of True and False Pterygium, or Other Growths at the Sclerocorneal Junction. PMID- 16692307 TI - A New Operation for Capsulo-muscular Advancement, Combined with Partial Resection. PMID- 16692308 TI - Sclerocorneal Trephining for Glaucoma. Complications and Failures in One Hundred Cases. PMID- 16692309 TI - Trephine Operation for Glaucoma-Late Infection from an Acute Conjunctivitis. PMID- 16692310 TI - Report of a Case of Detachment of the Retina Occurring in a Case of Neuroretinitis, Relieved by Scleral Trephining Operation, Associated with Incision of the Choroid and Retina. No Recurrence After a Period of Eight Months. PMID- 16692311 TI - Report of a Series of Extractions of Cataract in the Capsule After Subluxation with the Capsule Forceps. PMID- 16692312 TI - Preliminary Capsulotomy in the Operation for the Extraction of Immature Cataract. PMID- 16692313 TI - A New Double-disc Ophthalmoscope, with Independent Driving Gears, and Improved Electric Light Attachment. PMID- 16692314 TI - Cranial Deformity Associated with Ocular Changes. PMID- 16692315 TI - Melanosis of the Conjunctiva. Report of a Case. PMID- 16692316 TI - Acquired Symmetrical Opacities of the Cornea of Unusual Type. PMID- 16692317 TI - Blepharochalasis-Report of Two Cases. PMID- 16692319 TI - Contraction of the Frontalis in Abduction of the Eyeball. PMID- 16692318 TI - A Case of Chronic Sporotrichosis of the Eye. PMID- 16692320 TI - A Case of Symmetrical Occlusion of the Pupils by the Development of Cysts and Small Solid Masses from the Uveal Layer of the Iris. PMID- 16692321 TI - The Pathology of the Corneal Section and Its Complications in Cataract Extraction. PMID- 16692322 TI - Psammosarcoma of the Orbit in a Girl of Thirteen. Successful Removal with Preservation of the Eyeball and Its Functions. PMID- 16692323 TI - Radium Therapy in a Case of Orbital Metastasis Consequent Upon an Intraocular Sarcoma. PMID- 16692324 TI - Tuberculosis of the Bulbar Conjunctiva. PMID- 16692325 TI - Report of a Case of Tuberculosis of the Sclera of Probable Primary Origin. PMID- 16692326 TI - A Case of Alternating Transient Monocular Blindness Ending in Complete Loss of Vision in the Left Eye. PMID- 16692327 TI - Retinitis with Massive Exudation. PMID- 16692328 TI - The Convergence Index as a Measure of the Converging Power. PMID- 16692330 TI - Massive Granuloma of the Sclera (Brawny Scleritis), with the Report of an Unusual Case: Pathologic Examination by Dr. F. H. Verhoeff. PMID- 16692329 TI - Curvature and Index Myopia, with Report of Cases. PMID- 16692331 TI - Salvarsan in Interstitial Keratitis and in Optic Nerve Lesions. PMID- 16692332 TI - The Ives-Langdon Photometer. PMID- 16692333 TI - A Review of the History of the Electric Ophthalmoscope, with a Demonstration of a New Ophthalmoscope, which Consists of a Rearrangement of the Parts of Antecedent Instruments. PMID- 16692334 TI - A Simple Modification of the Berger Loupe. PMID- 16692335 TI - Nagel's Anomaloscope for Testing Color Vision. PMID- 16692336 TI - The May Electric Ophthalmoscope. PMID- 16692337 TI - Stronger Eye Magnets. PMID- 16692338 TI - A More Exact Method of Measuring Fields of Vision and Arcs of Rotation. PMID- 16692339 TI - Ocular Phenomena Accompanying Three Cases of Gastro-Intestinal Disorder. PMID- 16692340 TI - Lumbar Puncture and Examination of the Spinal Fluid in Affections of the Eye. PMID- 16692341 TI - Tuberculous Meningitis with Special Reference to the Tubercle of the Choroid and Its Pathologic Manifestations. PMID- 16692342 TI - Malignant Uveitis Treated with Thyroid Extract. PMID- 16692343 TI - Progressive Essential Atrophy of All Layers of the Iris. PMID- 16692345 TI - Pemphigus of the Conjunctiva.-Report of Two Cases. PMID- 16692344 TI - The Ophthalmias of Palestine. PMID- 16692346 TI - Report of a Case of Colloid Degeneration of the Upper Lid. PMID- 16692347 TI - A Case of Epibulbar Sarcoma. PMID- 16692348 TI - Multiple Melanotic Nevi of Conjunctiva, with Malignant Invasion of the Corneal Limbus. PMID- 16692349 TI - Report of Two Cases of Metastatic Carcinoma of the Choroid and One Case of Myxosarcoma of the Orbit. PMID- 16692350 TI - A Case of Primary Intradural Tumor of the Optic Nerve. PMID- 16692351 TI - A Primary Tumor of the Optic Nerve Successfully Removed, with Preservation of the Eyeball, by the Kronlein Method. PMID- 16692352 TI - A Contribution to the Subject of Tumors of the Eyelid and Orbit. PMID- 16692354 TI - Family Degeneration of the Macula Lutea, with a Suggestion as to Its Cause. PMID- 16692353 TI - Hereditary Posterior Polar Cataract, with Report of a Pedigree. PMID- 16692355 TI - Retinal Disease with Massive Exudation and an Allied Condition: Report of Two Cases. PMID- 16692356 TI - Spontaneous Reattachment of the Retina after Twenty-two Years' Duration; Report of Case with Remarks. PMID- 16692357 TI - A Review of the Glaucoma Question, with Special Reference to Newer Conclusions of Its Etiology and Departures of Its Surgical Treatment. PMID- 16692358 TI - Anomalies of the Accommodation Clinically Considered. PMID- 16692359 TI - Nervous Symptoms from Contusions of the Eye. PMID- 16692360 TI - A Report of Several Cases of Chronic Simple Glaucoma Treated by Iridotasis. PMID- 16692361 TI - Tenotomy of the Inferior Oblique Muscle. PMID- 16692362 TI - Rosacea Keratitis and Certain Other Forms of Marginal Keratitis, Neuropathic in Origin. Treatment by Pericorneal Neurotomy. PMID- 16692363 TI - Some Experiences in Enucleation under Local Anesthesia. PMID- 16692364 TI - Three Years' Experience in Sclerocorneal Trephining in Glaucoma. PMID- 16692365 TI - The Operative Treatment of Partial Staphyloma of the Cornea and of Fistula of the Cornea with a Conjunctival Flap; Conjunctivo-keratoplasty (Kuhnt). PMID- 16692366 TI - The Ambulant After-treatment of Cataract Extraction; with a Note on Post operative Delirium and on Striped Keratitis. PMID- 16692367 TI - Extraction of Cataracts in the Capsule by a Slight Modification of the von Graefe Method. PMID- 16692368 TI - Improved Capsule Forceps for Intracapsular Cataract Extractions. PMID- 16692369 TI - The Etiology of Iritis. PMID- 16692370 TI - A Case of Blepharochalasis-A Variety of Dermatolysis. PMID- 16692371 TI - Observations upon Herpes Corneae "Febrilis," with Reference, Especially, to Etiology. PMID- 16692372 TI - Two Patients with Unusual Corneal Opacities. PMID- 16692373 TI - Ophthalmoscopic Work in the British Base Hospitals. PMID- 16692374 TI - Report of the Removal of a Tumor at the Apex of the Orbit with Preservation of the Eyeball in a Case of Plexiform Neuroma of the Eyelid. Microscopic Examination. PMID- 16692375 TI - An Unusually Large Osteoma of the Frontal, Ethmoidal, and Sphenoidal Sinuses Involving the Orbit and Anterior Cerebral Fossa, Presenting no Subjective Symptoms Other than Proptosis of the Eyeball. PMID- 16692376 TI - Two Cases of Orbital Growth-(A) Mixed Tumor of the Lacrimal Gland. (B) Small Round-cell Sarcoma in an Infant. PMID- 16692377 TI - Recurrent Sarcomata at the Limbus Treated by "Electric Desiccation". PMID- 16692378 TI - An Epibulbar Melanosarcoma of the Limbus which Completely Disappeared under X-ray Treatments. PMID- 16692379 TI - Report of a Case of Melanosarcoma of the Orbit Treated with Radium. PMID- 16692380 TI - A Practical Lamp for Ophthalmic Operations. PMID- 16692381 TI - A Uniform Visual Test-object. PMID- 16692382 TI - A Case of Syphilitic Retinochoroiditis Juxtapapillaris, with Microscopic Examination. PMID- 16692383 TI - Tuberculosis of the Retinal Vessels. PMID- 16692385 TI - Report of Cases of Quinin Amblyopia, with Remarks. PMID- 16692384 TI - A Clinical and Pathological Study of Two Cases of Miliary Tubercles of the Choroid. PMID- 16692386 TI - Retrobulbar Neuritis with Central Scotoma from Toxic Action of Thyroidin. PMID- 16692387 TI - Optic Neuritis from Syphilitic Leptomeningitis. PMID- 16692388 TI - Cyanosis Retinae et Conjunctivae in Connection with Pulmonary Stenosis and Patent Ductus Arteriosus. PMID- 16692390 TI - High Hyperopia from a Clinician's Standpoint. PMID- 16692389 TI - A Case of Pituitary Body Disease with Reference to the Effects of Early Operation. PMID- 16692391 TI - Unusual Complications in a Case of Glaucoma after Atropin. PMID- 16692393 TI - Isolated Paralysis of the Cervical Sympathetic Accompanying Goiter. PMID- 16692392 TI - A Case of Botulism. PMID- 16692394 TI - Paralysis of Divergence. PMID- 16692395 TI - Divergent Concomitant Strabismus, the Muscular Anomalies and Refractive Errors Accompanying It, and Its Treatment. PMID- 16692396 TI - Color Interlacing and Perimetry. PMID- 16692398 TI - The Motais Operation for Ptosis. PMID- 16692397 TI - A Discussion of Some Newer Principles in Dealing with Uncomplicated Cataract. PMID- 16692399 TI - A Study of Hemorrhage into the Anterior Chamber Subsequent to Operations for Hard Cataract. PMID- 16692401 TI - The Electromagnet and the Magnet Operation in Ophthalmic Surgery. PMID- 16692402 TI - A Case of Aneurysm of the Internal Carotid Artery (Intracranial Portion) and Its Effect Upon the Patient's Vision. PMID- 16692400 TI - The Etiology of Retinitis Circinata. PMID- 16692403 TI - Trypanosome Keratitis-An Experimental Study. PMID- 16692404 TI - Necrology: Dr. Wilbur Boileau Marple. PMID- 16692405 TI - Five Years' Experience with Iridotasis. PMID- 16692406 TI - Late Infection Following an Operation of Iridotasis for Chronic Glaucoma. Enucleation. PMID- 16692407 TI - Glaucoma as a Result of Herpes Zoster Frontalis, with Report of Cases. PMID- 16692408 TI - Report of a Case of Primary Sarcoma of the Iris: with Remarks. PMID- 16692409 TI - On Varieties of the Fatigue of Accommodation as Registered by the Ergograph. PMID- 16692410 TI - Asteroid Hyalitis. PMID- 16692411 TI - Quantitative Perimetry; Practical Devices and Errors. PMID- 16692412 TI - Sarcoma of the Choroid with Secondary Changes. PMID- 16692413 TI - Report of a Case of Bilateral Inferior Hemianopsia. PMID- 16692414 TI - Neuroma Embryonale of Left Lateral Ventricle of Brain with Double Choked Disc. PMID- 16692415 TI - Recurrent Neuroepithelioma. PMID- 16692416 TI - A Case of Cavernous Angioma of the Orbit. PMID- 16692417 TI - Penetrating Injury Limited to the Eyeball Followed by Acute Tetanus. PMID- 16692418 TI - A Case of Unilateral Proptosis. Exploratory Operation. Tentative Diagnosis of an Ethmoid Osteoma. PMID- 16692419 TI - A Note on the Relation of Sex Linkage to Certain Hereditary Diseases of the Eyes. PMID- 16692420 TI - A Case Showing Multiple Congenital Abnormalities of the Eye; the Origin of the Vitreous Indicated by One of Them. PMID- 16692421 TI - Pupillary Studies, with Special Reference to Anisocoria. PMID- 16692423 TI - Report of Committee on Standardization of the Illumination of Test-Cards and Perimeters. PMID- 16692422 TI - Heterochromia Iridis, Heterochromic Cyclitis, and Allied Conditions. PMID- 16692424 TI - Two Cases of Congenital Coloboma of Iris and Choroid. PMID- 16692425 TI - The Removal of Foreign Bodies From Within the Eyeball and Orbit: Report of Cases. PMID- 16692426 TI - The Treatment of Detachment of the Retina, with Special Reference to Muller's Resection of the Sclera. PMID- 16692427 TI - Report of a Tumor of the Frontal Lobe of the Brain with Ocular Symptoms. Autopsy. PMID- 16692428 TI - Report of a Case of Total Loss of Vision in One Eye and Partial Loss in the Other Completely Relieved by a Sellar Decompression Operation. PMID- 16692429 TI - Concerning the Etiology of Hereditary Optic Nerve Atrophy. Report of Two Cases with Interesting X-ray Findings. PMID- 16692430 TI - Tuberculin in Diseases of the Eye. PMID- 16692431 TI - Some Ocular Conditions Affecting the Efficiency of the Aviator. PMID- 16692432 TI - The Inertia of Adjustment of the Eye for Clear Seeing at Different Distances. PMID- 16692433 TI - On the Construction of Test-Objects, Especially Those for Measuring Fatigue of Accommodation. PMID- 16692434 TI - The Trial-Frame for Test-Lenses, with Demonstration of a New Model. PMID- 16692435 TI - An Improved System of Illumination for the Electric Ophthalmoscope. PMID- 16692436 TI - Some Observations on the Restoration of the Orbital Socket. PMID- 16692437 TI - Report of Four Cases of Orbital Tumors Successfully Removed, with Preservation of Vision, Through an Orbital Incision Along the External Orbital Rim. PMID- 16692438 TI - Report of Two Orbital Tumors. PMID- 16692440 TI - Ophthalmoscopic Conditions Simulating Sarcoma of the Choroid: (a) Granuloma of the Choroid. (b) Tumor Formation Associated with Retinitis with Massive Exudation. PMID- 16692439 TI - Tumor of the Orbit, with Extension to Frontal and Temporal Regions, Treated by X rays. PMID- 16692441 TI - Report of a Case of Transient Left Homonymous Hemianopsia. PMID- 16692442 TI - A Practical Point in the Technic of Corneal Tattooing, the Value of Which Is Not Commonly Recognized. PMID- 16692443 TI - Hypermetropia Responsible for Heterophoria, Astigmatism, and Myopia. PMID- 16692444 TI - Conservative Treatment of Asthenopia in Children. PMID- 16692445 TI - Hereditary Congenital Ptosis: With Report of 64 Cases Conforming to the Mendelian Rule of Dominance. PMID- 16692447 TI - Tuberculous Iridokeratitis-Some Features in Its Pathology. PMID- 16692446 TI - A Consideration of the Causes of Heterochromia Iridis, with Special Reference to a Paralysis of the Cervical Sympathetic. PMID- 16692448 TI - A Study of the Conjunctival Circulation, to Determine Early Signs of Arteriosclerosis. PMID- 16692450 TI - The Thermophore in Ophthalmic Practice. PMID- 16692449 TI - The Vitreous and Its Replacement After Loss. PMID- 16692451 TI - The Present Status of Dental Sepsis in Its Relation to Certain Ocular Disorders. PMID- 16692452 TI - Cavernous Sinus Thrombosis, with Notes of Five Cases. PMID- 16692453 TI - Hyperplastic Exudative Retinitis-Non-Hemorrhagic. PMID- 16692454 TI - Solid Edema of the Face: Report of Three Cases. PMID- 16692455 TI - Histologic Findings in Eyeballs Lost Through Complications Following Sclerocorneal Trephining. PMID- 16692456 TI - Cataract in Retinitis Pigmentosa; Its Pathology and Treatment. PMID- 16692457 TI - Microscopic Findings in a Case of Coralliform Cataract, with Remarks on Congenital Cataracts in General. PMID- 16692458 TI - Permanent Vascular Changes Following Injuries to the Eye. PMID- 16692459 TI - A Case of Evulsion of the Optic Nerve. PMID- 16692460 TI - Ocular Affections Dependent Upon Disease of the Tonsils: Report of Two Cases. PMID- 16692461 TI - Acute Retrobulbar Neuritis of Septic Origin, Resulting in Rapid and Complete Loss of Vision. PMID- 16692462 TI - Primary Treatment of War Injuries of the Lids and Orbits. PMID- 16692463 TI - Penetrating and Perforating Wounds of the Eyeball with Diagnosis and Treatment of Retained Foreign Bodies. PMID- 16692464 TI - Necrology: Richard Andrews Reeve, B.A., M.D., LL.D. PMID- 16692465 TI - Concerning the Ocular Phenomena in the Psychoneuroses of Warfare. PMID- 16692466 TI - Trachoma in the Camps and Hospitals of the Army: A Brief Survey of the Subject from a Military Point of View. PMID- 16692467 TI - Ocular Complications Due to Typhoid Inoculations. PMID- 16692468 TI - Further Experience in the Ocular Functions of Aviators. PMID- 16692469 TI - Research Work in Ophthalmology at the Medical Research Laboratory of the Air Service Division of the Surgeon General's Office and its Bearing on the Teaching of Ophthalmology in America. PMID- 16692470 TI - A Test for the Judgment of Distance. PMID- 16692471 TI - The Maddox Rod Screen Test. PMID- 16692472 TI - A Contribution to the Study of Dark-adaptation. PMID- 16692473 TI - Notes on a Popular Presentation of Specimens Showing the Migration of the Eye of the Flounder and the Imperfect Eyes of Cave Fauna. PMID- 16692474 TI - War Injuries of the Eyelids: Plastic Operations for a Few Types. Case Reports with Photographs and Drawings. PMID- 16692475 TI - Introductory Remarks. Exhibition of Film, "Fit to Fly". PMID- 16692476 TI - Multiple Vaccination of the Eyelids. PMID- 16692477 TI - A Case of Keratitis Profunda (or Disciformis?) with Microscopic Examination. PMID- 16692478 TI - Report of Two Unusual Complications of Herpes Zoster Ophthalmicus: (a) Acute Retrobulbar Neuritis (Neuritis Axialis); (b) Acute Glaucoma. PMID- 16692480 TI - Coloboma and So-called Congenital Dislocation of the Lens. PMID- 16692479 TI - Some Cases of Lethargic Encephalitis. PMID- 16692481 TI - A Case of Multiple Dermoids of the Eyeball with Other Anomalies of Development. PMID- 16692482 TI - Tuberculosis of the Retina. PMID- 16692483 TI - Subretinal Exudate Simulating Sarcoma of the Choroid, with Anatomic Examination. PMID- 16692485 TI - Visual Acuity at Low Illumination-Apparatus and Results. PMID- 16692484 TI - Atypical Disseminated Choroiditis Due to Disease of the Accessory Sinuses. PMID- 16692486 TI - A Stereomicrometer-An Instrument of Precision for Measuring Stereopsis. PMID- 16692488 TI - German Field Optical Outfit. PMID- 16692487 TI - A Six-Meter Stereoscope. PMID- 16692489 TI - Non-Operable Tumor of the Orbit and Brow, Treated Successfully with Radium. Report of a Case with a Few Remarks. PMID- 16692490 TI - A Positive Focal Tuberculin Reaction in a Spindle-Cell Sarcoma which Had Perforated the Sclera. PMID- 16692491 TI - Clinical and Pathologic Report of a Case of Intraocular Tumor. PMID- 16692492 TI - Two Cases of Epithelioma of the Eyeball and Lids. Operation. No Return after Six Years. PMID- 16692493 TI - Report of Three Additional Cases of Paralysis of Accommodation Due to Focal Infections. PMID- 16692494 TI - Accommodation in the Lensless Eye Occurring in a Woman Aged Seventy-three Years, Following Extraction of Cataract. PMID- 16692495 TI - Requirements and Experiences in Ocular Examinations for the Army. PMID- 16692496 TI - Persistent Posterior Fibrovascular Sheath of the Lens. A Report of Two Clinical Cases, and Three Bulbs Examined Microscopically. PMID- 16692497 TI - Antidiphtheritic Serum in Severe Ocular Infections, with Special Reference to Hypopyon Keratitis. PMID- 16692499 TI - The Eyes of the Signalman. PMID- 16692498 TI - The Eye in Hereditary Syphilis. PMID- 16692500 TI - Remarks on Diagnosis and Treatment of Luetic Involvement of the Optic Pathways. PMID- 16692501 TI - Military Ophthalmology. PMID- 16692502 TI - Notes on Vernal Conjunctivitis. PMID- 16692503 TI - A Brief History of the Formation of the Division of Head Surgery, with Special Reference to Ophthalmology. PMID- 16692505 TI - Tuberculous Meningitis. PMID- 16692504 TI - Ocular Manifestations Following Exposure to Various Types of Poisonous Gases. PMID- 16692506 TI - Visual Field Findings in a Case of Brain Tumor. PMID- 16692507 TI - The Eye and the Endocrine System. PMID- 16692508 TI - An Unusual Manifestation of Tuberculous Keratitis. PMID- 16692510 TI - Thermophore. PMID- 16692509 TI - Argyrol. PMID- 16692511 TI - Holder for Half-Curved Needle. PMID- 16692512 TI - Optical Prescription Blank. PMID- 16692513 TI - The Campperimeter-An Illuminated Perimeter with Campimeter Features. PMID- 16692515 TI - Uveitis Dependent upon Focal Infection in the Appendix. PMID- 16692514 TI - Neuropathic Keratitis Setting in Long after Alcoholic Injections into Gasserian Ganglion for Relief of Trigeminal Neuralgia. PMID- 16692516 TI - (1) Epibulbar Carcinoma Developing Nine Years after the Removal of a Papilloma of the Corneoscleral Margin; (2) Small, Spindle-celled Hemangiosarcoma of the Eyelid of a Child Aged Five Months, Excision Followed by Radium Treatment; (3) Pathologic Histology of a Concussioned Eye Following Gun-shot Wound of Orbit. PMID- 16692517 TI - Bilateral Glioma: Report of a Case Unsuccessfully Treated with Radium. PMID- 16692518 TI - Glioma Retinae. PMID- 16692519 TI - A Case of Melanosarcoma of the Choroid. PMID- 16692521 TI - Successful Removal of the Orbital Portion of the Optic Nerve for a Primary Intradural Tumor, with Preservation of the Eyeball and Motility, by the Kronlein Method. PMID- 16692520 TI - A Case of Intra-ocular Tumor with Unusual Operative Complications. PMID- 16692522 TI - Factors which Influence the Color Sensitivity of the Peripheral Retina. PMID- 16692523 TI - A Case of Accommodation in a Lensless Eye. PMID- 16692524 TI - Prolonged Monocular Occlusion as a Test for the Muscle Balance. PMID- 16692525 TI - Hereditary Optic Atrophy. PMID- 16692526 TI - Eye Tables for the Determination of Compensations from Loss to the Earning Ability of a Person from Damages to the Functions of One or Both Eyes. PMID- 16692527 TI - Cyclopia, its Bearing upon Certain Problems of Teratogenesis and of Normal Embryology; with a Description of a Cyclocephalic Monster. PMID- 16692528 TI - Mooren's Ulcer of the Cornea: A Histologic Report. PMID- 16692529 TI - Carcinoma of the Orbit, with the Report of a Case. PMID- 16692530 TI - Increase of Hyperopia in Diabetes. PMID- 16692531 TI - Medical-Social Service and Follow-Up Work in the Eye Hospital. PMID- 16692532 TI - Visual Disturbance in Hysteria. PMID- 16692533 TI - Total Keratoplasty. PMID- 16692534 TI - Thrombosis of Retinal Vein after Influenza. PMID- 16692535 TI - Recurrent Retinal Hemorrhages of Adolescence with Report of a Second Case. PMID- 16692536 TI - Three Unusual Cases of Foreign Bodies in and about the Eyeball. PMID- 16692537 TI - Capsulotomy by a New Procedure. PMID- 16692538 TI - Results Obtained with Muller's Resection of the Sclera in Detachment of Retina Due to High Myopia. PMID- 16692539 TI - Nasolacrimal Surgery in Ophthalmologic Perspective. PMID- 16692540 TI - The Insertions of the Ocular Muscles as seen in Text-Books and in the Dissecting Room. PMID- 16692541 TI - The Action of the Obliques and the Bearing of Head-Tilting in the Diagnosis of Paralysis. PMID- 16692542 TI - A Case of Intermittent Exophthalmos. PMID- 16692543 TI - Double Luxation of the Eyeballs in a Case of Exophthalmic Goiter. PMID- 16692544 TI - Loss of Vision from Sympathetic Inflammation with Recovery Following the Use of Tuberculin. PMID- 16692545 TI - Loss of Vision in One Eye Restored by Pituitary Feeding in a Case of Compensatory Pituitary Hypertrophy. PMID- 16692546 TI - The Estimation of Compensation in Disability Resulting from Accidental Loss of Visual Acuity. PMID- 16692548 TI - A Modified Accommodation Line and Prince's Rule. PMID- 16692547 TI - An Unusual Case of Conjunctival Irritation. PMID- 16692549 TI - Illuminated Eye Spud with Magnifier. PMID- 16692550 TI - Foreign-Body Spud Illuminator. PMID- 16692551 TI - Forceps for Tendo-Muscle Lengthening. PMID- 16692552 TI - Sections of Coralliform Cataract, and a Microscopic Specimen of Vitreous Humor from Asteroid Hyalitis. PMID- 16692553 TI - Tonometric Chart. PMID- 16692554 TI - A Case of Mooren's Ulcer. PMID- 16692555 TI - Sarcoma of the Cornea: Case Report. PMID- 16692556 TI - Melanosarcoma (Melanoma) of the Choroid Occurring in Brothers. PMID- 16692557 TI - Report of a Case of Primary Tubular Epithelioma of the Lacrimal Sac. PMID- 16692558 TI - Glioma Retinae Treated by X-Rays, with Apparent Destruction of the Tumor and Preservation of Normal Vision. PMID- 16692559 TI - The Use of Radium Plugs in the Dissolution of Orbital Gliomatous Masses Developing after Excision of the Globe. PMID- 16692560 TI - Coats' Disease of the Retina: Report of Two Cases. PMID- 16692561 TI - Lipaemia Retinalis. PMID- 16692562 TI - Some Impressions Derived from the Study of Recurrent Hemorrhages into the Retina and Vitreous of Young Persons. PMID- 16692563 TI - The Ocular Symptoms of Epidemic Encephalitis. PMID- 16692564 TI - The Effect of Variations in Intensity of Illumination on Acuity, Speed of Discrimination, Speed of Accommodation, and Other Important Eye Functions. PMID- 16692565 TI - Some Hitherto Unrecognized Signs in Skiascopy. PMID- 16692566 TI - The Ocular Changes in Infantile Scurvy: Report of a Case. PMID- 16692567 TI - The Relation of Headache to Functional Monocularity. PMID- 16692569 TI - The Character of Iritis Caused by Focal Infection. PMID- 16692568 TI - Three Cases of Word-Blindness. PMID- 16692570 TI - Suggestions for a Uniform Method of Estimating Loss of Visual Efficiency Following Industrial Eye Injuries. PMID- 16692571 TI - Some Features in the Technic of Trephining the Cornea for the Relief of Glaucoma. PMID- 16692572 TI - The Compensation Problem in Ophthalmology. PMID- 16692573 TI - Repeated Operations for Glaucoma: Report of Case. PMID- 16692574 TI - Friability of the Iris a Factor in Iridectomy for Hypertension. PMID- 16692575 TI - The Action of Adrenalin on the Glaucomatous Eye. PMID- 16692577 TI - After-Cataract. PMID- 16692576 TI - The Physiologic Mode of Action of Mydriatics and Miotics, Explaining Their Effects in Hypertension (Glaucoma). PMID- 16692578 TI - Intracapsular Cataract Extraction by Traction Alone. PMID- 16692579 TI - Oil Cyst of Orbit Removed by Kronlein's Operation. PMID- 16692580 TI - Tuberculoma of the Orbital Cavity. Exhibition of Photograph. Literature. PMID- 16692581 TI - Basal Cell Carcinoma of the Orbit and Ethmoid: Operation; Radium Application. PMID- 16692582 TI - Studies in Monocular and Binocular Accommodation, with Their Clinical Application. PMID- 16692583 TI - Simple Methods of Orthoptic Training. PMID- 16692584 TI - Correction of Squint by Muscle Recession with Scleral Suturing. PMID- 16692585 TI - Epidemic Encephalitis from the Standpoint of the Ophthalmologist. PMID- 16692586 TI - Ophthalmoplegia Exterior, with Report of Cases. PMID- 16692587 TI - The Place of Ophthalmology in the Undergraduate Medical Curriculum. PMID- 16692588 TI - Observations, Extending Over a Period of Three Months, in a Case of Lipaemia Retinalis. PMID- 16692589 TI - Retinitis Proliferans of Both Eyes Due to Secondary Anemia Produced by Hodgkin's Disease: Report of a Case. PMID- 16692590 TI - An Ophthalmic Illuminator. PMID- 16692591 TI - Fusion Tubes and Monoscope. PMID- 16692592 TI - A Convenient Pocket Retinoscope. PMID- 16692593 TI - A Tilting Trial-Frame. PMID- 16692595 TI - Demonstration of the Development of the Human Eye. PMID- 16692594 TI - A Demonstration Eye. PMID- 16692596 TI - Panophthalmitis Following Infection Through a Cystoid Cicatrix. PMID- 16692597 TI - Sympathetic Inflammation Due to Faulty Trephining. PMID- 16692598 TI - An Unusual Epibulbar Carcinoma. PMID- 16692599 TI - Luetic Ophthalmitis. PMID- 16692600 TI - Altitudinal Hemianopsia-Occlusion of the Inferior Temporal Artery. PMID- 16692601 TI - Need of Protecting the Eyes in Our Industrial Schools as Illustrated by a Case. PMID- 16692602 TI - A Technic for Producing Experimental Ocular Tuberculosis in Animals. PMID- 16692603 TI - Cyst of the Iris with a Study of the Container and Its Contents. PMID- 16692604 TI - The Diagnostic and Therapeutic Use of Uveal Pigment in Injuries of the Uveal Tract and Sympathetic Ophthalmia. PMID- 16692605 TI - Ocular Manifestations in a Case of Hypophyseal Syphilis. PMID- 16692606 TI - Case of Hypophyseal Disease Probably of Syphilitic Origin. PMID- 16692607 TI - A Study of the Sugar Tolerance in Aglycosuric Patients with Cataract. PMID- 16692608 TI - A Modified Keratome Incision for Iridectomy in Glaucoma. PMID- 16692609 TI - Bridge Coloboma of the Iris-Slit Lamp Examination of Two Cases. PMID- 16692610 TI - Concerning the Use of Diphtheria Antitoxin in Post-Operative Infections and Acute Uveal Disease. PMID- 16692611 TI - Factors Influencing the Choice of Method for Cataract Extraction. PMID- 16692612 TI - Multicolor Tattooing of the Cornea. PMID- 16692613 TI - Contribution to the Chemicopharmaco-Dynamic Relationship of Atropin and Homatropin. PMID- 16692614 TI - Transitory Visual Disturbances Observed During the Course of Occipital Lesions, and Their Significance with Respect to Cerebral Localization. PMID- 16692616 TI - Melanotic Epibulbar Tumor Dispelled by the Use of Radium. PMID- 16692615 TI - Central Scotoma in Anterior Uveitis: A Further Contribution. PMID- 16692617 TI - Recurrent Pigmented Growth at Limbus. PMID- 16692618 TI - A Case of Large Round-Cell Sarcoma of the Upper Eyelid. PMID- 16692619 TI - Variations in the Primary Insertions of the Ocular Muscles. PMID- 16692620 TI - Glioma of Retina. PMID- 16692621 TI - Thermophore. PMID- 16692622 TI - Tuberculosis of the Eye. PMID- 16692623 TI - Siderosis Bulbi. PMID- 16692624 TI - A Case of Extensive Tuberculosis of the Eyeball. PMID- 16692625 TI - Persistent Hyaloid Artery. PMID- 16692626 TI - Osteoma of the Orbit. PMID- 16692627 TI - A Case of Adenocarcinoma Probably Arising From the Lacrimal Gland. PMID- 16692628 TI - A Case of Encapsulated Angioma of the Orbit. PMID- 16692629 TI - Posterior Transillumination of the Eyeball. PMID- 16692630 TI - Selective Thermotherapy. PMID- 16692631 TI - Malignant Disease of the Retina. PMID- 16692632 TI - Autopsy Report of a Case of Bilateral Glioma Retinae with Gliomatous Involvement of the Iris. PMID- 16692634 TI - Supplementary Notes on Lipaemia Renitalis: Effects of Insulin. PMID- 16692633 TI - Siderosis Bulbi with Dilated Inactive Pupil. Recovery of Pupillary Activity After Removal of Foreign Body. PMID- 16692635 TI - The Significance of Deep Ocular Pain in Retinal Detachment. PMID- 16692636 TI - Bilateral Detachment of the Retina in two Successive Pregnancies. PMID- 16692637 TI - Observations on Phacoanaphylactic Endophthalmitis. PMID- 16692638 TI - Monocular Diplopia, with Especial Reference to that Associated with Cerebral Lesions. PMID- 16692639 TI - Metastatic Choroiditis from Focal Infection. PMID- 16692640 TI - Optic Atrophy Manifested by Visual Disturbance Following Distant Hemorrhage. PMID- 16692642 TI - Examination of the Blind Spot of Mariotte. PMID- 16692641 TI - Traumatic Neurology of the Eye in Relation to Cerebral and Other Injuries. PMID- 16692643 TI - The Effect of General Tuberculosis on the Eye, with a Report of Frequently Overlooked Symptoms. PMID- 16692645 TI - Report of Committee on Undergraduate Teaching of Ophthalmology. PMID- 16692644 TI - Some of the Essentials in Legislation for the Relief of the Blind. PMID- 16692646 TI - Report of Committee on Graduate Teaching of Ophthalmology. PMID- 16692647 TI - Hydrophthalmos (Buphthalmos or Infantile Glaucoma)-Report of a Case with Microscopic Findings. PMID- 16692648 TI - Report of Two Experiences with Acute Glaucoma. PMID- 16692649 TI - Iridectomy in Glaucoma: A New Technic. PMID- 16692650 TI - An Operation for Blepharoptosis, with the Formation of a Fold in the Lid. PMID- 16692651 TI - Duct Conservation in Lacrimal Abscess. PMID- 16692652 TI - A Report of the Changes Found in the Eyes of Rabbits Following the Injection of Living Tubercle Bacilli into the Common Carotid Artery. PMID- 16692654 TI - The Advantages of Complete Discission by the V-Shaped Method. PMID- 16692653 TI - Serum and Lens-Antigen Extract Treatment for the Prevention and Cure of Cataract: With the Report of the Results Obtained in One Hundred and Thirty-one Cataracts. PMID- 16692655 TI - Glioma of the Retina with Deposits of Glioma Cells on the Iris and Other Ocular Tissues. PMID- 16692656 TI - Concerning Extensive Orbital Cavernous Hemangioma, in Association with Pulsating Exophthalmos. Enucleation of Eye and Complete Exenteration of Orbit, Mainly by Electro-Coagulation. PMID- 16692657 TI - Blunt Keratome for Making Section for Cataract Extraction. PMID- 16692659 TI - Suction Tip for Eye Work. PMID- 16692658 TI - A Simple Lighting System for the Tangent Screen. PMID- 16692660 TI - Scissors for Magnet Extension for Extraction of Foreign Bodies from the Eye. PMID- 16692661 TI - Diaphragm to Attach to the Shahan Lamp for Use in Skiascopy. PMID- 16692662 TI - Scissors Tip for Magnet. PMID- 16692663 TI - The Corneal Splitter. PMID- 16692664 TI - Protective Goggles. PMID- 16692665 TI - Pemphigus of the Conjunctiva. PMID- 16692667 TI - Facsimile of Carmalt Testimonial. PMID- 16692666 TI - The Role of the Epithelial Cell in Conjunctival and Corneal Infections. PMID- 16692668 TI - Inflammatory Swellings Simulating Dacryocystitis. PMID- 16692669 TI - Case of Gumma of the Tear-Sac. PMID- 16692670 TI - Intradermal Test with Uveal Pigment in Traumatic Iridocyclitis. PMID- 16692671 TI - Senile External Exudative Macular Retinitis with Remarks on Similar Tumor Formation in a Case of Angioid Streaks. PMID- 16692672 TI - Some Lid Tumors. PMID- 16692673 TI - Glioma of the Retina. PMID- 16692674 TI - Pathologic Report on Two Cataractous Eyes. PMID- 16692675 TI - Glioma of the Iris. PMID- 16692676 TI - Glioma of the Retina with Deposits of Glioma Cells on the Iris and Other Ocular Tissues. PMID- 16692677 TI - Clinical Contribution to the Etiology of Dystrophia Epithelialis Corneae. PMID- 16692678 TI - Two Cases of Traumatic Keratitis in the New-Born. PMID- 16692679 TI - Lantern Demonstration of Some Vitreous Changes as Seen with the Diaphragm Lamp and Corneal Microscope. PMID- 16692680 TI - Asteroid Hyalitis-Report of a Case in Which the Bodies Developed While under Observation. PMID- 16692681 TI - A Case of Epibulbar Polymorphous Sarcoma. PMID- 16692682 TI - Pituitary Growth with Marked Improvement in Field and Vision Following Its Removal. PMID- 16692683 TI - A Case of Central Homonymous Hemianopic Scotoma. PMID- 16692684 TI - Abrupt Monocular Blindness Resulting in Complete Optic Atrophy During Encephalitis with Double Papillitis: X-Ray of Optic Canals. PMID- 16692685 TI - Thermophore Therapy and Experimental Studies. PMID- 16692686 TI - Chronic Juvenile Glaucoma: Report of a Case. PMID- 16692688 TI - A Collective Study of the Treatment of Congenital Hydrophthalmos. PMID- 16692687 TI - The Extraction of Senile Cataract by Vacuum Fixation. PMID- 16692689 TI - Report of the Committee on Undergraduate Teaching of Ophthalmology. PMID- 16692690 TI - Report of the Committee on Graduate Teaching of Ophthalmology. PMID- 16692691 TI - The Knapp Adrenalin Mydriasis Reaction in Direct Descendants of Patients with Primary Glaucoma. PMID- 16692692 TI - Radiation of the Cervical Sympathetic and Other Measures to Reduce Glaucomatous Tension. PMID- 16692693 TI - The Filtration Angle as Concerned in Iridectomy for the Relief of Non Inflammatory Glaucoma. PMID- 16692694 TI - Progress of Senile Cataract under Hygienic Care. PMID- 16692695 TI - Globular Masses on the Pupillary Margin in Acute Circumscribed Chorioretinitis. Clinical and Pathologic Study. AB - 1. A group of cases is described in which inflammatory lesions in the posterior segment of the eye were associated with the deposit of cells on the margin of the pupil and the posterior surface of the cornea.2. The path of wandering cells in relation to localized inflammatory lesions in the eye is discussed.3. The pathologic findings of a case of chorioretinitis at the optic disc are described.4. An explanation is offered for the sector-formed defect in the visual field of such cases. PMID- 16692696 TI - Separation of the Choroid, with Report of a Spontaneous Case. PMID- 16692697 TI - Corneal Growth Followed by Abscess. PMID- 16692698 TI - Ossification of Detached Retina and Choroid. PMID- 16692699 TI - Case for May Ophthalmoscope. PMID- 16692701 TI - Anatomic Method of Recording Fields in Perimetry. PMID- 16692700 TI - Medullated Nerve-Fibers Involving the Macula. PMID- 16692702 TI - The Use of Blood Serum in the Treatment of Gonorrheal Ophthalmia with Report of a Case. PMID- 16692704 TI - Subnormal Accommodation. PMID- 16692703 TI - A Case of Dystrophia Epithelialis Corneae. PMID- 16692705 TI - The Response of the Lens Capsules in the Act of Accommodation. PMID- 16692706 TI - Some Unusual Corneal Cases as Seen with the Slit-Lamp. PMID- 16692707 TI - The Percentage Evaluation of Macular Vision. PMID- 16692708 TI - Primary Sarcoma of the Eyelids in Children, with Report of Two Cases Treated with Radium. PMID- 16692709 TI - Orbital Cellulitis Following Hordeolum. PMID- 16692710 TI - A Case of Lymphangioma of the Orbit. PMID- 16692711 TI - The Field of Fixation; Introducing a New Technique. PMID- 16692713 TI - The Etiology and Treatment of Endogenous Iritis: An Analysis of 100 Consecutive Cases in Private Practice. PMID- 16692712 TI - Treatment of Simple Chronic Glaucoma. PMID- 16692714 TI - Neoplasms of the Lacrimal Gland with Report of Three Cases. PMID- 16692715 TI - Intra-Ocular Hypertension with Deep Anterior Chamber; Report of Two Cases with Microscopic Findings. PMID- 16692716 TI - An Analysis of 4,541 Ocular Injuries, Being a Contribution to Industrial Ophthalmology. PMID- 16692717 TI - Acute Dacryoadenitis Caused by Focal Infection. PMID- 16692718 TI - The Pathologic Histology of the Lacrimal Sac in Chronic Purulent Dacryocystitis. PMID- 16692720 TI - Indications for the Use of Pilocarpin in Dislocation of the Crystalline Lens. A Preliminary Report. PMID- 16692719 TI - Glaucoma in Myopic Eyes. PMID- 16692721 TI - Concerning the Ocular Symptoms of Erythremia (Chronic Polycythemia Vera), with Special Reference to the Fundus Picture. PMID- 16692722 TI - Glaucoma. PMID- 16692723 TI - Concerning a Law to Lessen Hereditary Blindness. PMID- 16692724 TI - Report of a Case of Tumor of the Optic Nerve. PMID- 16692725 TI - Report of a Patient with Incomplete Ophthalmoplegia. PMID- 16692726 TI - The Clinical Features and Etiologic Factors of Essential Progressive Atrophy of the Iris and the Formation of Holes in Its Tissue. A Second Communication. PMID- 16692727 TI - The Pelvic Organs as Foci of Infection in Inflammatory Diseases of the Eye. PMID- 16692729 TI - An Effective Treatment for Sympathetic Uveitis. PMID- 16692730 TI - Advantages of Plano Trial-Lenses. PMID- 16692728 TI - Evolution of the Pathogenesis of Sympathetic Ophthalmia, with Report of an Unusual Case. PMID- 16692731 TI - A Case of Plagiocephaly. PMID- 16692732 TI - A Case of Thrombosis of the Central Vein of the Optic Nerve with Subsequent Glaucomatous Manifestations. PMID- 16692733 TI - Familial Maculocerebral Degeneration in Half-Brother and Sister. PMID- 16692734 TI - Congenital Grouped Pigmentation of the Retina. PMID- 16692735 TI - Hyperphoria in Near Work. PMID- 16692736 TI - Suspension of the Fusion Impulse as a Therapeutic Measure. PMID- 16692737 TI - Studies in Ocular Fatigue. II. Convergence Fatigue in Practice. PMID- 16692738 TI - The Ocular Complications of Lipodystrophia Progressiva. PMID- 16692739 TI - An Analysis of the Marks Obtained in the Different Subjects by the Candidates for the Certificate of the American Board for Ophthalmic Examinations. PMID- 16692740 TI - Evaluation of Visual Efficiency: The Basic Factor in Computing Industrial Compensation for Loss of Vision. PMID- 16692741 TI - Congenital Aniridia. PMID- 16692742 TI - Metastatic Carcinoma of the Choroid and of the Iris: A Study of the Disease, with a Report of the Clinical History of a Case, and an Investigation of Its Pathologic Anatomy. PMID- 16692743 TI - Luetic Interstitial Keratitis of Traumatic Origin. PMID- 16692744 TI - Appendix: Report of Committee on Compensation for Eye Injuries , Section on Ophthalmology, American Medical Association. PMID- 16692746 TI - Corneoconjunctival Suture for Cataract Operations. PMID- 16692745 TI - A Large Exostosis of the Orbit Successfully Removed by Operation. PMID- 16692747 TI - The Surgical Treatment of Wounds of the Cornea with Prolapsed Iris. PMID- 16692748 TI - A Case of Hemangiomatous Endothelioma. PMID- 16692749 TI - Freely Movable Pigmented Cyst in the Anterior Chamber. PMID- 16692750 TI - Observations on Glaucoma in Morgagnian Cataract. PMID- 16692751 TI - Further Studies on the Light Sense in Early Glaucoma. PMID- 16692752 TI - Anterior Ocular Tuberculosis: A Group of Cases Followed Over a Period of Years. PMID- 16692753 TI - A Preliminary Report of Three Cases of Keratoconus. PMID- 16692754 TI - A Photographic Study of Holes in the Macular Region and Some Other Associated Changes. PMID- 16692755 TI - Glaucoma in Generalized Vascular Nevus of the Skin. Report of Case with Angiomatous Changes in the Iris. PMID- 16692756 TI - Sclero-post-iridectomy, a Rational Operation in Glaucoma-Report of a Case. PMID- 16692757 TI - Detachment of Retina Following Trephining for Glaucoma, with Complete Re attachment. PMID- 16692759 TI - Concerning the Site of Angioid Streaks of the Fundus Oculi. PMID- 16692758 TI - Angioid Streaks. A Report Concerning Two Cases. PMID- 16692760 TI - Angiomatosis Retinae, a Clinical Report of One Case. PMID- 16692761 TI - Air Embolism of the Retinal Vessels. PMID- 16692762 TI - Parinaud's Conjunctivitis with Suppuration in the Pre-auricular, Submaxillary, and Supraclavicular Glands. PMID- 16692763 TI - A Case of Endothelioma of the Orbit. PMID- 16692764 TI - Hensen and Voelckers's Experiments on the Mechanism of Accommodation; an Interpretation. PMID- 16692765 TI - Diphtheria Antitoxin in Large Doses for Sympathetic Ophthalmia. PMID- 16692766 TI - Observations on Central, Paracentral, and Caecocentral Scotomas. PMID- 16692767 TI - Comparative Chemical Studies of the Ocular Fluids, of Cerebrospinal Fluid, and of the Blood. PMID- 16692768 TI - An Experimental Study of the Local Action of Podophyllin upon Eyes, with a Report of a Clinical Case. PMID- 16692769 TI - A Plea for the Establishment of Schools of Ophthalmology. PMID- 16692770 TI - Graft of Tenon's Capsule for Perforation of the Flap in Trephining Operation. PMID- 16692771 TI - An Adjustable Tangent Screen with Artificial Daylight Illumination. PMID- 16692772 TI - Moving Pictures for the Demonstration of Certain Ocular Conditions. PMID- 16692773 TI - Simplification of the Principles of Ophthalmic Optics. PMID- 16692774 TI - The Retinitis of Malignant Hypertension. PMID- 16692775 TI - Tuberculous Iridocyclitis as a Cause of the Heterochromia of Fuchs. PMID- 16692776 TI - Conjunctivo-plasty in Certain Corneal Affections. PMID- 16692777 TI - A Function of Blood in Corneal Vascularization. PMID- 16692778 TI - A Corneo-sclero-conjunctival Suture in Operations for Cataract. PMID- 16692779 TI - A New Operation for Removing Cataracts with Their Capsules. PMID- 16692780 TI - Wounds of the Eye by Projectiles of Small Caliber or Low Velocity. PMID- 16692781 TI - Removal of a Number Six Lead Shot from within the Eyeball by Specially Devised Forceps with the Aid of the Double Plane Fluoroscope. PMID- 16692782 TI - Melanosis Oculi with the Report of a Case. PMID- 16692783 TI - Ocular Disease, Notably of the Uveal Tract, Induced by Streptococcus Viridans Infection. PMID- 16692784 TI - Slit-lamp Study of an Unusual Case of Congenital Cataracts. PMID- 16692785 TI - Cyst of the Uveal Layer of Iris at Pupillary Margin in a Case of Posterior Synechiae with Secondary Glaucoma. PMID- 16692786 TI - Report upon a Case of Free Cyst in the Anterior Chamber with Exhibition of Patient. PMID- 16692787 TI - The Regeneration of Aqueous. PMID- 16692788 TI - Bilateral Obstruction of the Central Artery of the Retina. PMID- 16692789 TI - The Photographic Diagnosis of Some Punctate Retinal Conditions. PMID- 16692790 TI - Retinal Degeneration in Macular Region without Cerebral Symptoms. PMID- 16692791 TI - Contribution to the Pathologic Anatomy of Siderosis Bulbi. PMID- 16692792 TI - Hemangiomas of Cerebellum and Retina (Lindau's Disease): With the Report of a Case. PMID- 16692793 TI - The Value of Moving Pictures in Ophthalmologic Teaching: A Preliminary Report. PMID- 16692794 TI - Disc-like Degeneration of the Macula with Microscopic Report Concerning a Tumor like Mass in the Macular Region. PMID- 16692795 TI - Remarks on the Artificial Induction of Ocular Hypertension by Compression of the Jugular Veins. PMID- 16692796 TI - Chronic Progressive External Ophthalmoplegia: Report of a Case with Necropsy. PMID- 16692797 TI - Test-letters Which Comply with the Physiologic Requirements of a Visual Test object. PMID- 16692798 TI - Frequency of the Claude Bernard-Horner Syndrome: Report of 16 Cases. PMID- 16692799 TI - The Intravenous Use of Typhoid-Paratyphoid Vaccine in Diseases of the Eye. PMID- 16692800 TI - A Case of Melanotic Sarcoma of the Choroid with Probable Cerebral Metastases. PMID- 16692801 TI - Pathologic Sections of a Case of Essential Atrophy of the Iris. PMID- 16692802 TI - A Muscle Tucking Instrument. PMID- 16692803 TI - Test-types. PMID- 16692804 TI - Choroidal Melanomas. PMID- 16692805 TI - The Aqueous Humor.-II: A Critical and Experimental Study of Its Formation and Chemical Composition. PMID- 16692806 TI - Posterior Lenticonus. PMID- 16692807 TI - Congenital Pigmentation of the Cornea (Krukenberg Type). PMID- 16692808 TI - A Study of the Comparative Anatomy of the Extra-ocular Muscles. PMID- 16692809 TI - Pigmented Lesions of the Conjunctiva. PMID- 16692810 TI - A Critical Study of the Modern Ophthalmoscope: Contributions to Its Construction and Use. PMID- 16692811 TI - Detachment of Descemet's Membrane. PMID- 16692812 TI - Biochemical Studies of the Blood in Patients with Senile Cataract. PMID- 16692813 TI - A Case of Sarcoma of the Iris. PMID- 16692814 TI - Histologic Findings in a Case of Localized Tuberculous Chorioretinitis. PMID- 16692815 TI - The Nature of So-called Koeppe Nodules. PMID- 16692817 TI - A Case of Mixed Tumor of the Lacrimal Gland. PMID- 16692816 TI - Tuberculoma Corneae. PMID- 16692818 TI - A Case of Trichinosis with Exophthalmos. PMID- 16692819 TI - On the Operation for Shrunken Cataracts in Adolescents. PMID- 16692820 TI - A Portable Adaptometer. PMID- 16692821 TI - The Relationship of the Flavobacterium Ophthalmiae to Periodic Ophthalmia in Horses. PMID- 16692822 TI - Normal Vision, Nine Years After the Passage of a Steel Chip Through the Crystalline Lens. PMID- 16692823 TI - Cyst of the Vitreous: Report and Presentation of a Case. PMID- 16692824 TI - Crystalline Deposits in the Vitreous, with a Report of Two Cases of Calcium Soaps: Studied Microscopically and Histo-Chemically. PMID- 16692825 TI - Microchemical and Histological Findings in a Case of Asteroid Hyalitis. PMID- 16692826 TI - Homologous Retinoblastoma in Identical Twins. PMID- 16692827 TI - A Retinoblastoma in a Man Aged Forty-eight Years. PMID- 16692828 TI - Terminal Stage in a Case of Retinitis with Massive Exudation. PMID- 16692830 TI - Ocular Lesions in Fetal Syphilis. PMID- 16692829 TI - Lipemia Retinalis in a Diabetic. PMID- 16692831 TI - Bilateral Coloboma of the Optic Nerve Associated with Holes in the Disc and a Cyst of the Optic Sheath. PMID- 16692832 TI - The Role of Intracranial Aneurysms in the Production of Ocular Palsies. PMID- 16692833 TI - Prismoscopic Perimetry of the Macular Area: A New Method of Mapping Out Paracentral Scotomas and Other Defects in the Visual Fields. PMID- 16692834 TI - Divergence Excess: Its Incidence, Its Correlation with Refraction, and the Value of Orthoptic Treatment. PMID- 16692835 TI - Exhibition of New Apparatus: A Tangent Screen with Mirror Fixation to be used on the Ferree-Rand Perimeter. PMID- 16692836 TI - Tenotomy of the Inferior Oblique. PMID- 16692838 TI - A Contribution to the Study of Phlyctenular Kerato-Conjunctivitis. PMID- 16692837 TI - Clinical and Experimental Studies on Traumatic Annular Opacity of the Anterior Lens Surface (Vossius' Ring). PMID- 16692839 TI - Tissue Culture in Ophthalmic Research. PMID- 16692840 TI - Botulism and Its Ophthalmic Lesions as Determined by Research with the Toxin of Clostridium Botulinum. PMID- 16692841 TI - The Anatomy and Histology of the Eye and Orbit of the Rabbit. PMID- 16692842 TI - Report of a Case of Epibulbar Basal Cell Carcinoma with Penetration of the Globe. PMID- 16692843 TI - A Case of Complete Exenteration of the Orbit for Neglected Epithelioma of the Eyelids by Means of Electrocautery. PMID- 16692844 TI - Exactly Appositional Sutures in the Cataract Operation. PMID- 16692845 TI - Etiology and Treatment of Keratoconus. PMID- 16692847 TI - Allergic Phenomena in the Course of Tuberculous Keratitis: With a Report of Two Cases. PMID- 16692846 TI - Dendritic Keratitis following Therapeutic Inoculation of Malaria. PMID- 16692848 TI - Peripheral Pigmentation of the Cornea Associated with Lenticular Degeneration of the Pseudo-Sclerosis Type. PMID- 16692849 TI - Report of a Case of Nodular Scleritis with Observations as to Etiology and Pathology of this Disease. PMID- 16692850 TI - Benign Melanoma and Melano-Epithelioma of the Choroid. PMID- 16692851 TI - Primary Sarcoma of the Iris. Removed by Iridectomy. No Recurrence in Three and a Half Years. PMID- 16692852 TI - Absorption by Radium of Intra-Ocular Granulation following Stab Wound at the Corneal Limbus. PMID- 16692853 TI - Report of Small Tumor of the Iris which Caused Secondary Glaucoma-Microscopic Examination-Remarks on Relation of Melanoma to Sarcoma of the Iris. PMID- 16692854 TI - Secondary Glaucoma with Detached Retina in both Eyes. PMID- 16692855 TI - Some Observations in Regard to Glaucoma Simplex. PMID- 16692856 TI - Sight Saving Class Work from the Standpoint of the American Ophthalmological Society and the National Society for the Prevention of Blindness. PMID- 16692857 TI - Cystic Degeneration of the Retina. PMID- 16692859 TI - The Permeability of the Lens Capsule to Water, Dextrose, and Other Sugars. PMID- 16692858 TI - Microscopic Observations in a Case of Retinitis Pigmentosa. PMID- 16692860 TI - Concerning Lamellar Membranes of the Anterior Surface of the Lens. PMID- 16692861 TI - Changes in Refraction with Presbyopia. PMID- 16692862 TI - The Acuity of Binocular Depth Perception in Hemianopsia. PMID- 16692863 TI - Ocular Hazards and Safeguards. PMID- 16692864 TI - Monochromatic Vision. PMID- 16692865 TI - Some Ocular Lesions in Septicemia. PMID- 16692866 TI - Report of a Case of Transplantation of the Human Cornea. PMID- 16692867 TI - Two Per Cent. Epinephrin Solutions as Substitutes for Laevo-Glaucosan. PMID- 16692868 TI - A Lens Expressor. PMID- 16692869 TI - Changes in the Visual Fields as shown by Moving Pictures. PMID- 16692870 TI - An Investigation of the Sugar Content of the Ocular Fluids Under Normal and Abnormal Conditions, and the Glycolytic Activity of the Tissues of the Eye. PMID- 16692871 TI - Peri-Papillary Detachment of the Retina Accompanying Papilledema. PMID- 16692872 TI - Slit-Lamp Observations on Keratoconus. PMID- 16692873 TI - Avertin as an Anesthetic in Ophthalmic Surgery. PMID- 16692874 TI - Removal of Glass from Anterior Chamber. PMID- 16692876 TI - Exophthalmos Associated with Diabetes Insipidus and Large Defects in the Skull Bones. PMID- 16692875 TI - Late Post-Operative Separation of the Choroid. PMID- 16692877 TI - Diphtheria of the Conjunctiva. PMID- 16692878 TI - Nodular Opacity of the Cornea. PMID- 16692879 TI - The Cataract of Post-Operative Tetany with Report of Three Cases. PMID- 16692881 TI - Intrascleral Nerve Loops. PMID- 16692880 TI - Concerning Cyclodialysis in Simple Glaucoma. PMID- 16692882 TI - The Circulation of the Aqueous-Preliminary Report. PMID- 16692884 TI - A Report of an Experimental Study of the Reparative Process of the Lens Capsule after Injury. PMID- 16692883 TI - The Organ Specific Properties and Antigenic Power in the Homologous Species of Alpha Crystallin. PMID- 16692885 TI - Concerning a Membrane between the Vitreous and the Anterior Chamber, Seen after Removal of the Crystalline Lens and Its Capsule. PMID- 16692886 TI - The Condition of the Ocular Structures Immediately after Removal of the Lens in Capsule, as Determined by Microscopic Examination. PMID- 16692887 TI - The Anterior Vitreous-in Health and Disease. PMID- 16692888 TI - Ultraviolet Light Ocular Therapy. PMID- 16692889 TI - Further Notes on the Metabolism of the Retina. PMID- 16692890 TI - Epipapillary Tissues. PMID- 16692891 TI - Measurements of Retinal Vessels in Early Hypertension. PMID- 16692893 TI - Some Cases of Separation of the Retina and Its Subsequent Behavior in Non-Myopic Eyes. PMID- 16692892 TI - The Presence of Vitamin A in the Retina. PMID- 16692894 TI - Analysis of the Proliferative and Exudative Elements in Some Cases of Sympathetic Ophthalmia. PMID- 16692895 TI - Analogies and Differences of the Second and Eighth Nerves and End-Organs: Generalizing, Preliminary, Anatomic Considerations, Especially with Reference to Choked Disc, Glaucoma, and Choked Labyrinth. PMID- 16692896 TI - Blindness and Papilledema in Guernsey Calves, Usually Bulls, Including the Results of Postmortem Examination of Two of the Affected Animals. PMID- 16692897 TI - Papilledema Due to Disease of the Sphenoidal Sinus. PMID- 16692898 TI - Measles Encephalitis: A Clinical Report of Some Eye Findings. PMID- 16692899 TI - Albinism in the Progeny of Negro and White Parents. PMID- 16692900 TI - Exophthalmos in Infantile Scurvy. PMID- 16692901 TI - Subnormal Accommodation. PMID- 16692902 TI - Mixed Tumor of the Lacrimal Gland. PMID- 16692903 TI - Retinal Perivascular Delineation. PMID- 16692904 TI - Retrobulbar Sarcoma. PMID- 16692905 TI - Cyst of the Anterior Chamber. PMID- 16692906 TI - Vein Communicating with Schlemm's Canal. PMID- 16692907 TI - Cilium in the Vitreous. PMID- 16692908 TI - Nodular Dystrophy of the Cornea. PMID- 16692909 TI - Tuberculous Chorioretinitis. PMID- 16692910 TI - Rents in the Retina. PMID- 16692911 TI - Ocular Lesions Caused by Intracranial Hemorrhage. PMID- 16692912 TI - Study of the Pathologic Changes in the Optic Nerves and Chiasm in Comparison with Changes in the Visual Field in Association with Large Pituitary Tumors. PMID- 16692913 TI - On the Origin and Disposition of the Cells in Endophthalmitis. PMID- 16692914 TI - An Investigation of the Ferment Activity of the Rabbit's Vitreous. PMID- 16692915 TI - The Histopathology of Bowman's Membrane. PMID- 16692916 TI - Tribromethanol (Tribromethyl-Alcohol, Avertin) as an Anesthetic in Eye Surgery. PMID- 16692918 TI - Instrument for the Exercise of the Extrinsic Ocular Muscles. PMID- 16692917 TI - Scotometry: History and Technique; with a Scotometric Tangent Screen and Scales. PMID- 16692919 TI - Suprarenin Bitartrate. PMID- 16692920 TI - Case of Complete Unilateral Traumatic Ophthalmoplegia, Externa and Interna. PMID- 16692921 TI - Congenital Absence of the Lacrimal Puncta and Canaliculi. PMID- 16692922 TI - The Pathologic Changes in the Orbit in Progressive Exophthalmos, with Special Reference to Alterations in the Extra-Ocular Muscles and the Optic Discs. PMID- 16692924 TI - Leptotrichosis Conjunctivae (Parinaud's Conjunctivitis): Artificial Cultivation of the Leptotriches in Three Out of Four Cases. PMID- 16692923 TI - Premonitory Lid Edema in the Typhoid Group. PMID- 16692925 TI - Endarteritis Obliterans with Spontaneous Gangrene of Both Corneas. PMID- 16692927 TI - Dystrophia adiposa corneae. PMID- 16692926 TI - Mooren's Ulcer. PMID- 16692928 TI - Additional Experiments Verifying the Presence of Treponema Pallidum in the Cornea in Experimental Interstitial Keratitis. PMID- 16692929 TI - An Unusual Corneal Lesion, Probably Congenital and Familial in Character: A Slit Lamp Study. PMID- 16692931 TI - Experimental Cataract in Vitamin G Deficiency. PMID- 16692930 TI - The Circulation of the Intra-Ocular Fluid: The Importance of the Optic Nerve. PMID- 16692932 TI - Significance of Specific Infiltration at the Site of Injury in Sympathetic Ophthalmia. PMID- 16692933 TI - Concerning Allergy and Immunity in Ocular Tuberculosis. PMID- 16692934 TI - The Standard Classified Nomenclature of Disease-Illustrated by the Classification of Heterophoria and Heterotropia. PMID- 16692935 TI - The Scotometry of Retinal Edema. PMID- 16692936 TI - Pantocain as a Local Anesthetic in Ophthalmology. PMID- 16692937 TI - The Light-Streaks on the Retinal Blood-Vessels. PMID- 16692938 TI - Venous Angioma of the Retina, Optic Nerve, Chiasm, and Brain: A Case Report with Postmortem Findings. PMID- 16692939 TI - Pigmentation of the Optic Nerve. PMID- 16692940 TI - On the Association of Sclerosis of the Cerebral Basal Vessels with Optic Atrophy and Cupping: Report of 10 Cases. PMID- 16692941 TI - Central Visual Field Studies in Intracranial Lesions. PMID- 16692942 TI - Dr. Elisha North, of New London, Connecticut, the Founder of the First Eye Infirmary in the United States. PMID- 16692943 TI - Some Essentials and Securities Which Stabilize Muscle Operations. PMID- 16692944 TI - Spud for Dissecting Conjunctiva from Sclerocorneal Margin in Trephine Operation. PMID- 16692945 TI - Unusual Case of Cyst of the Orbit. PMID- 16692946 TI - Condensed Vision Chart. PMID- 16692947 TI - Eye Pad for the Administration of Heat Treatments. PMID- 16692948 TI - On the Pathology of Bullous Keratitis. PMID- 16692949 TI - Ocular Manifestations of Raynaud's Disease. PMID- 16692950 TI - The Temporal Half-Moon. PMID- 16692951 TI - Orbital Actinomycosis. PMID- 16692952 TI - Concerning Tuberculous Lesions of the Retina. PMID- 16692953 TI - Concretions in Henle's Glands. PMID- 16692954 TI - Argyrosis of the Conjunctiva. PMID- 16692955 TI - Cyclodialysis Combined with Iridectomy in Glaucoma Simplex; a Preliminary Report. PMID- 16692956 TI - A Review of the Cases of Glaucoma Admitted to the Massachusetts Eye and Ear Infirmary Over a Period of Two Years. PMID- 16692957 TI - Evisceration for Hemorrhagic Glaucoma, Followed by Orbital Melanosarcoma and Metastases Resulting in Death. PMID- 16692958 TI - Congenital Cyst of the Retina. PMID- 16692959 TI - The Osmotic Equilibrium between Blood and Intra-Ocular Fluid as Influenced by Anisotonic Injections: Clinical Significance. PMID- 16692960 TI - Is the Aqueous Humor a Dialysate? PMID- 16692961 TI - The Respiratory Function of the Aqueous. PMID- 16692962 TI - A New Test for Visual Acuity. PMID- 16692963 TI - Enophthalmos in Horner's Syndrome. PMID- 16692964 TI - Solitary Neurofibroma of the Orbit. PMID- 16692965 TI - Sarcoma of Choroid in a Shrunken Globe Exhibiting Fibrous Formation about the Tumor. PMID- 16692966 TI - An Epithelial Cyst of the Posterior Chamber; Clinical History and Microscopic Anatomy of the Enucleated Eye. PMID- 16692967 TI - The Operative Treatment of Retinal Detachment with Electro-Coagulation. PMID- 16692968 TI - A Case of Tuberculous Sclerokeratitis. PMID- 16692969 TI - Sympathetic Scleritis. PMID- 16692970 TI - The Use of the Superior Oblique as an Internal Rotator in Third Nerve Paralysis. PMID- 16692971 TI - Membranous Conjunctivitis with a Persistent Membrane Completely Covering the Cornea. PMID- 16692973 TI - Presentation of Instruments and Appliances: Astigmatic Charts. PMID- 16692972 TI - Thrombosis of the Central Retinal Vein, with Special Reference to the Streptococcus Viridans. Recovery. PMID- 16692974 TI - Ulcus Serpens in Glaucoma. PMID- 16692975 TI - Sarcoma of the Iris. PMID- 16692976 TI - Holes in the Retina. PMID- 16692977 TI - Tuberculosis of the Eye. PMID- 16692978 TI - Laurence-Moon-Biedl Syndrome. PMID- 16692979 TI - Hereditary Optic Atrophy (Leber's Disease): A Comparative Review of the Literature and the Report of a Family. PMID- 16692981 TI - Congenital Anterior Staphyloma. PMID- 16692980 TI - Band Keratitis. PMID- 16692982 TI - Lesions of the Cornea Caused by Birth Trauma. PMID- 16692983 TI - Visual Hallucinations Accompanying Organic Lesions of the Brain, with Special Reference to Their Value as Localizing Phenomena. PMID- 16692984 TI - Visible Retinal Embolism. PMID- 16692985 TI - Routine Ophthalmoscopic Examination as an Aid in the Management of Maternity Cases. PMID- 16692986 TI - An Unusual Corneal Opacity: A Persistent, Spreading Opacity Following Herpes Zoster Corneae. PMID- 16692987 TI - Ocular Symptoms and Complications Observed in Dengue. PMID- 16692988 TI - Intra-Ocular Mycosis. PMID- 16692989 TI - Dystrophy of the Corneal Endothelium (Cornea Guttata), with Report of a Histologic Examination. PMID- 16692990 TI - On Hyphema after Cataract Extraction. PMID- 16692991 TI - Light Minimum in the Diabetic. PMID- 16692992 TI - Paralysis of the Superior Rectus Muscle. PMID- 16692993 TI - Tubercle-Like Nodules of Episclera and Eyelids, Bilateral. PMID- 16692994 TI - Dislocation of the Ring of Soemmering, Its Removal, with Some Notes on Its Pathology. PMID- 16692996 TI - Concerning Iridodialysis as a Clinical Entity, Its Surgical Treatment: Report of Cases. PMID- 16692995 TI - Hereditary Optic Atrophy (Leber's Disease). PMID- 16692997 TI - Salzmann's Nodular Corneal Dystrophy: Its Pathology and a Suggested Therapy. PMID- 16692998 TI - The Occurrence of Ciliary Processes on the Iris. PMID- 16692999 TI - Stigmatoscopy. PMID- 16693000 TI - Glaucoma Accompanying Nevus Flammeus. PMID- 16693001 TI - A Peculiar Contraction of the Visual Fields Associated with Syphilis of the Central Nervous System. PMID- 16693002 TI - Three Unusual Cases of Foreign Bodies in and about the Eyeball. PMID- 16693003 TI - Sympathectomy for Retinitis Pigmentosa. PMID- 16693004 TI - The Iris Inclusion Operations for Glaucoma, with Especial Reference to Iridotasis. PMID- 16693005 TI - Cycloduction. PMID- 16693006 TI - A Factor in the Production of Divergence Increase with Near Vision. PMID- 16693007 TI - Advancement of the Superior Oblique and Inferior Oblique Ocular Muscles. PMID- 16693009 TI - Surgery of the Rectus Muscles of the Eye: Selection of Operative Procedures by Differential Diagnosis. PMID- 16693008 TI - Orthoptic Training and Surgery in Hyperphoria and Hypertropia Combined with Lateral Deviations. PMID- 16693010 TI - Some Observations as to Indications for Advancement or for Kindred Operations. PMID- 16693011 TI - Inclusion Blennorrhea. PMID- 16693012 TI - The Pathogenesis of Some Intra-Ocular Osseus Tissue. True Metaplasia in the Eye. PMID- 16693013 TI - The Ocular Manifestations Observed in Eighteen Cases of Intracranial Adamantinoma. PMID- 16693014 TI - Unilateral Exophthalmos: Etiologic and Diagnostic Studies in Eighty-two Consecutive Cases. PMID- 16693015 TI - A Case of Chloroma with Orbital Involvement Locally Benefited by X-Ray Therapy. PMID- 16693016 TI - The Association of Optic Neuritis, Facial Paralysis, and Facial Hemiatrophy. PMID- 16693018 TI - Pathologic Changes in the Anterior Half of the Globe in Obstruction in the Central Vein of the Retina. PMID- 16693017 TI - Arachnodactyly (Dystrophia Mesodermalis Congenita, Typus Marfans. Marfan's Syndrome. Dolichostenomelie). PMID- 16693019 TI - Tuberculous and Streptococcic Retinal Hemorrhages. PMID- 16693020 TI - Stereoscopic Demonstration: Stereoscopic Photograph of Lacrimal Sac after Injection with Thorium Dioxid. PMID- 16693021 TI - Local Anesthesia in Ophthalmology. PMID- 16693022 TI - The Effect of Malaria upon Certain Syphilitic Conditions of the Eye. PMID- 16693024 TI - Control of Intra-Ocular Hypertension by Systemic Medication. PMID- 16693023 TI - The Visual Prognosis for Children with Congenital Nystagmus: A Statistical Study. PMID- 16693025 TI - Essential Shrinkage of the Conjunctiva in a Case of Probable Epidermolysis Bullosa Dystrophica. PMID- 16693026 TI - Ocular Manifestations of Congenital Syphilis and Treatment by Induced Hyperpyrexia. PMID- 16693028 TI - Schlemm's Canal: Its Anastomoses and Anatomic Relations. PMID- 16693027 TI - Angioid Streaks and Osteitis Deformans. PMID- 16693029 TI - Leprosy and Tuberculosis of the Eye. PMID- 16693030 TI - Friedreich's Ataxia Showing Transitional Symptoms of Marie's Disease (Familial Spinocerebellar Ataxia). PMID- 16693031 TI - The Medical Treatment of Senile Cataract: General and Constitutional; Specific and Local. PMID- 16693032 TI - Lipin Interstitial Keratitis. PMID- 16693033 TI - New and Practical Charts and Lighting for Testing Visual Acuity and Locating the Astigmatic Axis: Simple, Standardized, and Complying with Modern Requirements. PMID- 16693034 TI - A Kinetic Test for Stereoscopic Vision. PMID- 16693035 TI - Lipodystrophia Progressiva with Ocular Complications: Further Report. PMID- 16693036 TI - Hereditary Macular Degeneration. PMID- 16693037 TI - Bullous Keratitis: A Rational Therapy. PMID- 16693038 TI - The Effects of Mydriatics upon Intra-Ocular Tension. PMID- 16693040 TI - Tuberculosis of the Choroid Associated with Generalized Miliary Tuberculosis. PMID- 16693039 TI - The Treatment of Sarcoma of the Uveal Tract. PMID- 16693041 TI - Vitreous Detachment, Anterior Dialysis, and over the Optic Nerve a Tumor-like Mass Consisting of the Detached Retina. PMID- 16693043 TI - The Structure of the Vitreous. PMID- 16693044 TI - The Mechanism of the Fovea. PMID- 16693042 TI - Studies on the Crystalline Lens. V. The Nature of the Reducing Substances in the Lens. PMID- 16693045 TI - Cataracts Produced in Albino Rats on a Ration Containing a High Proportion of Lactose or Galactose. PMID- 16693046 TI - Chronic Postoperative or Post-Traumatic Retinitis (Retinitis Serosa). PMID- 16693047 TI - Epithelial Downgrowth into the Anterior Chamber Following Cataract Extraction Arrested by Radium Treatment. PMID- 16693048 TI - Detachment of the Choroid after Cataract Extraction: Clinical and Experimental Studies, with Report of Seventy-five Cases. PMID- 16693049 TI - The Results of Cataract Extraction Five Years and More after Operation. PMID- 16693050 TI - An Analysis of 1,343 Intracapsular Cataract Extractions by 48 Operators Following the Verhoeff Method. PMID- 16693051 TI - The Effect of Bacterial Lysate in Staphylococcic Keratoconjunctivitis in Rabbits. PMID- 16693053 TI - The Diagnosis and Differentiation of Alternating Types of Concomitant Squint. PMID- 16693052 TI - Further Note on the Inclusion Bodies in Ophthalmia Neonatorum. PMID- 16693054 TI - Metastatic Ophthalmia in a Case of Pneumonia. Bacteriologic Findings. PMID- 16693055 TI - A Curved Keratome for Sclerectomy Operations. PMID- 16693056 TI - Luminous Dial. PMID- 16693057 TI - Multiple Maddox Rod with a Prism. PMID- 16693058 TI - Simplified Ophthalmoscope and Retinoscope as an Accessory to a Vest-Pocket Flashlight. PMID- 16693059 TI - A New Model of an Enophthalmometer and Exophthalmometer. PMID- 16693060 TI - A Binocular Microscope for Delicate Surgery. PMID- 16693061 TI - A Practical Method to Test the Strength of Ophthalmic Magnets. PMID- 16693063 TI - A Case of Tumor of the Antrum Involving the Eye. PMID- 16693062 TI - Pupil Goggles. Electrodes for Diathermic Heat. Galvanic Unit. PMID- 16693064 TI - The Talking Book. PMID- 16693066 TI - Filamentary Keratitis. PMID- 16693065 TI - The Conduction of Heat to the Interior of the Eye and Orbit from a Constant Temperature Applied to the Cornea. PMID- 16693067 TI - Experimental Uveitis: "Interference" Effect of Parenteral Administration of Proteins on Sensitization of the Uveal Tract. PMID- 16693068 TI - Theories of Color Sensation, the Phenomena of Simultaneous Contrast, and the Contrast Box. PMID- 16693069 TI - Diathermic and Galvanic Treatment of Separated Retina, Especially of the Flat Type, with Specially Devised Minimal Stop and Single and Multiple Platinum Micro Electrodes. New Magazine Containers. PMID- 16693070 TI - Papilledema, with Special Reference to Papilledema Associated with Sinus Disease. PMID- 16693071 TI - Convalescent Blood for Herpes Zoster. PMID- 16693072 TI - Retinal Periphlebitis in Septic Endophthalmitis and Its Ophthalmoscopic Picture. PMID- 16693073 TI - The Origin of the Brille in Crotalus Confluentus Lutosus (Great Basin Rattlesnake). PMID- 16693074 TI - Mathematical Observations on Jackson's Cross-Cylinder in Relation to the Conoid of Sturm. PMID- 16693075 TI - Complicated After-Cataract: Its Surgical Treatment. PMID- 16693076 TI - Subjective Studies of the Blind-Spot and Visual Fields. PMID- 16693077 TI - Central Visual Fields and Pupils in Neurosyphilis. PMID- 16693078 TI - Homonymous Hemianopia: Some Practical Points in Its Interpretation, with a Report of Forty-nine Cases in Which the Lesion in the Brain Was Verified. PMID- 16693079 TI - Diphtheria of the Conjunctiva: Two Cases. PMID- 16693080 TI - Ocular Muscle Palsies in a Case of Toxic Goiter. PMID- 16693081 TI - Polarized Light Attachment for Ophthalmoscopy. PMID- 16693082 TI - Choroidal Chorionepithelioma Secondary to Teratoma of the Testicle. PMID- 16693083 TI - Cavernous Hemangioma of the Choroid: Report of Five Cases in the Registry of Ophthalmic Pathology, Army Medical Museum, Washington, D. C. PMID- 16693084 TI - Bilateral Metastatic Carcinoma of Choroid-Successful X-Ray Treatment of One Eye. PMID- 16693086 TI - The Complications of the Forceps Intracapsular Cataract Operation, Based on an Analysis of 500 Successive Cases. PMID- 16693085 TI - Plexiform Neurofibromatosis (von Recklinghausen's Disease) Involving the Choroid, Ciliary Body, and Other Structures. PMID- 16693087 TI - Further Observations on Detachment of the Choroid After Cataract Extraction. PMID- 16693089 TI - Humoral Biopathology and the Eye: Some Inferences and Implications. PMID- 16693088 TI - Ocular Lesions Associated with Postoperative and Gestational Nutritional Deficiency. PMID- 16693090 TI - Sudden Occlusion of Retinal Arteries: Correlated Fundus Photographs and Fields in Branch Closures. PMID- 16693091 TI - A Study of the Changes in the Optic Discs, Visual Fields, and Vision Following Bisection of the Optic Chiasm. PMID- 16693092 TI - Chronic Traumatic Subdural Hematoma as a Cause of Choked Disc. PMID- 16693093 TI - Dermoid Cysts of the Orbit. PMID- 16693094 TI - Neo-Synephrin: Some Uses and Effects in Ophthalmology. PMID- 16693095 TI - A New Instrument for Facilitating the Suturing of the Wound in Operations for Cataract. PMID- 16693096 TI - Transilluminator. PMID- 16693097 TI - Capsule Forceps for Intracapsular Cataract Extraction. PMID- 16693098 TI - Dacryocystorhinostomy. PMID- 16693099 TI - The Reticulo-Endothelial System of the Eye. PMID- 16693101 TI - Experimental Iritis: The Ocular Reactions in Rabbits Sensitized to Streptococcus Viridans. PMID- 16693100 TI - Eye Changes in Epidemic Cerebrospinal Meningitis: A Clinical and Pathologic Study of 200 Cases. PMID- 16693103 TI - A Second Group of Arachnodactyly Cases. PMID- 16693102 TI - Ophthalmia Neonatorum: A Study of 261 Cases. PMID- 16693104 TI - Statistical Study of Spectral Hue Discrimination in Trichromatic and Dichromatic Eyes. PMID- 16693105 TI - Twinning and Ocular Pathology, with a Report of Bilateral Macular Coloboma in Monozygotic Twins. PMID- 16693106 TI - Anomalous Ophthalmic Artery with Eye Symptoms. PMID- 16693107 TI - Leiomyoma of the Iris: Report of a Case. PMID- 16693109 TI - Detachment of the Retina: Report on Operative Results in 164 Cases. PMID- 16693108 TI - A Case of Hyperplasia of the Epithelium of the Ciliary Processes (Ciliary Adenoma of Treacher Collins). PMID- 16693110 TI - Etiology of Idiopathic Retinal Detachment. PMID- 16693111 TI - Neuromyelitis Optica: A Report of Two Cases. PMID- 16693112 TI - Divergence Insufficiency: A Clinical Study. PMID- 16693113 TI - Relationship of Heterophoria to Divergence and Convergence, Based on Clinical Measurements. PMID- 16693114 TI - The Surgical Management of Ptosis, with Special Reference to the Use of the Superior Rectus Muscle. PMID- 16693115 TI - Tenon's Capsule Transplantation in Surgery of the Ocular Muscles, with Especial Reference to Postoperative Deviations with Adhesions between the Muscles and the Eyeball. PMID- 16693116 TI - The Mechanism of Formation of the Aqueous. PMID- 16693117 TI - Ocular Changes Experimentally Produced in Thyroparathyroidectomized Dogs, with Special Reference to Intra-Ocular Tension and Blood-Pressure: A Preliminary Report. PMID- 16693118 TI - Birth Injuries of the Cornea and Allied Conditions. PMID- 16693119 TI - Benzedrine in Refraction: Preliminary Report. PMID- 16693120 TI - Concerning the Intra-Ocular Invasion by the Larva of the Ascaris: Report of a Case with Unusual Complications. PMID- 16693121 TI - The Treatment of Ocular Tuberculosis. PMID- 16693122 TI - Preretinal Connective-Tissue Formation in the Vitreous in Acute Choroiditis: Report of Three Cases. PMID- 16693123 TI - The Pathogenesis of Disciform Degeneration of the Macula. PMID- 16693124 TI - The Genesis of the Cyclitic Membrane. PMID- 16693125 TI - The Neurogenic Origin of Choroidal Sarcoma. PMID- 16693126 TI - Inflammatory Pseudotumor of the Orbit: Report of a Case. PMID- 16693127 TI - A Cyst of the Posterior Chamber with a Microscopic Study of the Eye. PMID- 16693128 TI - Posterior Sclerotomy as a Form of Treatment in Subchoroidal Expulsive Hemorrhage. PMID- 16693129 TI - Stereoscopic Cards in Color for Children (Series A). PMID- 16693130 TI - Kindergarten Visual Acuity Chart. PMID- 16693131 TI - A Pendulum Scalpel. PMID- 16693132 TI - An Instrument for Demonstrating Simultaneous Color Contrast. PMID- 16693133 TI - Keratoplasty-Microscopic Study of the Corneal Grafts. PMID- 16693134 TI - Visual Field Studies in Asthenopia. PMID- 16693135 TI - Bilateral Metastatic Carcinoma of the Choroid. PMID- 16693137 TI - Precancerous Melanosis and Diffuse Malignant Melanoma of the Conjunctiva. PMID- 16693136 TI - A Micro-Camera for Visualization of Conjunctival Vessels. PMID- 16693138 TI - A Melanoma of the Iris with Pathologic Findings. PMID- 16693139 TI - Causes of Senile Cataract. PMID- 16693140 TI - Diabetes and Retinitis. PMID- 16693141 TI - Ocular Manifestations in Brucellosis (Malta Fever; Undulant Fever). PMID- 16693142 TI - Syphilitic Opticochiasmatic Arachnoiditis. PMID- 16693143 TI - Optic Tract Lesion, Probably the Result of Opticochiasmal Arachnoiditis Due to Infected Sphenoid Sinuses. PMID- 16693145 TI - A New Method for Transplanting Pterygium. PMID- 16693144 TI - Spastic Entropion Correction by Orbicularis Transplantation. PMID- 16693146 TI - The Value of Inferior Iridotomy in Operations for Cataract on Eyes with Posterior Synechiae or Pupillary Membrane. PMID- 16693147 TI - American Ophthalmology Grows Up: Turbulent Years, 1908-1915. PMID- 16693148 TI - Traumatic Retinal Angiopathy. PMID- 16693149 TI - Cultivation in Vitro of Human Conjunctival and Corneal Epithelium. PMID- 16693150 TI - Results of Autotransplantation of Cornea into the Anterior Chamber: Their Significance Regarding Corneal Nutrition. PMID- 16693151 TI - Some Physiologic and Pharmacologic Reactions of Isolated Iris Muscles. PMID- 16693152 TI - Aniseikonia. PMID- 16693154 TI - A Case of Aleukemic Lymphosis Involving the Upper Lids, with Pathologic Findings. PMID- 16693153 TI - Aniseikonia: A Study of 836 Patients Examined with the Ophthalmo-Eikonometer. PMID- 16693155 TI - The Newest Model of the Nordenson Zeiss Photographing Ophthalmoscope. PMID- 16693156 TI - Specially Designed Eye Sutures. PMID- 16693157 TI - A Contact Glass for Gonioscopy. PMID- 16693159 TI - Iridocapsulotomy Scissors. PMID- 16693158 TI - Prism Scale for Use at 50 Cm. PMID- 16693160 TI - Retinitis in Dermatomyositis. PMID- 16693162 TI - Retrograde Degeneration in the Optic Nerves and Retinal Ganglion Cells. PMID- 16693161 TI - The Biochemistry of the Optic Nerve. PMID- 16693163 TI - Notes on Experimental Keratitis: An Investigation of the Cellular Pathology with Particular Reference to the Macrophages. PMID- 16693164 TI - Shall We Use Cycloplegics? PMID- 16693165 TI - Subjective Study of Visual Aberrations. PMID- 16693166 TI - Mixed Cell Tumor of the Lacrimal Sac. PMID- 16693167 TI - Panophthalmitis and Sympathetic Ophthalmia. PMID- 16693168 TI - Metastatic Septic Endophthalmitis with Ring Abscess of the Cornea-Case Report, Clinical History, and Pathologic Anatomy. PMID- 16693169 TI - Orbital Hyperostosis in Meningioma. PMID- 16693170 TI - A Report of-(A) A Case of Leiomyoma of the Iris; (B) A Case of Hematoma of the Iris. PMID- 16693171 TI - Familial Progressive Juvenile Cataracts (Parathyroid Deficiency). PMID- 16693172 TI - A Family with Lattice Dystrophy of the Cornea. PMID- 16693174 TI - Psychosomatic Relationships and Their Therapeutic Implications in Glaucoma. PMID- 16693173 TI - Air Injection as a Factor in Maintaining Filtration Following Corneoscleral Trephining in Glaucoma. PMID- 16693175 TI - Field Changes After Satisfactory Filtration Operations for Glaucoma. PMID- 16693176 TI - Clinical Detection of Early Changes in the Visual Field. PMID- 16693177 TI - Final Ocular Result in a Case of Anterior Poliomyelo-encephalitis Twenty-seven Years After the Acute Attack. PMID- 16693178 TI - An Exposition of Some Round Macular Lesions. PMID- 16693179 TI - Certain Retinopathies Due to Changes in the Lamina Vitrea. PMID- 16693180 TI - Metastatic Carcinoma of the Iris. PMID- 16693181 TI - Multiple Myeloma with Bilateral Sixth Nerve Paralysis and Left Retrobulbar Neuritis. PMID- 16693183 TI - Six Cases of Lindau's Disease, Four Verified and Living. PMID- 16693182 TI - Keratomalacia and Cystic Fibrosis of the Pancreas. PMID- 16693184 TI - Plexiform Neurofibromatosis (von Recklinghausen's Disease) of the Orbit and Globe, with Associated Glioma of the Optic Nerve and Brain: Report of a Case. PMID- 16693185 TI - Evisceration of the Globe with Scleral Implant and Preservation of the Cornea. PMID- 16693186 TI - Historic Notes on Ophthalmoscopy. PMID- 16693187 TI - Accuracy Factors in Selective Thermotherapy. PMID- 16693188 TI - The Course of Certain Cases of Optic Atrophy with Cupping and Low Tension. PMID- 16693190 TI - Hyperphoria Tests Based Upon a New Principle. PMID- 16693189 TI - The Role of Ascorbic Acid (Vitamin C) in the Secretion of the Intra-Ocular Fluid. PMID- 16693191 TI - Two Rare Cases of Congenital Anomalies of the Eye with Clinical Implications. PMID- 16693192 TI - The Effect of Anoxemia on Dark Adaptation in the Normal and Vitamin A Deficient Subject. PMID- 16693193 TI - The Retina in Surgical Cases of Primary Hypertension. PMID- 16693194 TI - The Treatment of Trachoma with Sulfanilamide: A Report of 28 Cases. PMID- 16693195 TI - A Prism Bar of Thermoplastic Material for Measuring High Degrees of Squint. PMID- 16693196 TI - Tovell Eye Shield for Insensitive Cornea. PMID- 16693197 TI - Increasing the Magnetic Field Intensity of Portable Electromagnets. PMID- 16693198 TI - A Test for Binocular Vision Particularly Applicable to the Examination of Amblyopic Children (Modified Worth Test). PMID- 16693199 TI - A Test for Heterophoria. PMID- 16693200 TI - Improved Kinetic Test for Binocular Stereopsis. PMID- 16693201 TI - The Optical Principles of Telescopic Spectacles. PMID- 16693202 TI - Ocular Lesions of Boeck's Sarcoid. PMID- 16693203 TI - Coloboma of the Optic Nerve in the Human Embryo. PMID- 16693204 TI - The American Ophthalmological Society: A Retrospect of Seventy-five Years. PMID- 16693205 TI - Edward Delafield: A Sketch. PMID- 16693207 TI - An Investigation of Experimental Cataracts in the Albino Rat: Clinical Implications. PMID- 16693206 TI - After-Cataract. PMID- 16693208 TI - The Treatment of Inclusion Conjunctivitis with Sulfanilamide. PMID- 16693210 TI - Accidental Involvement of the Eyes in Vaccinia. PMID- 16693209 TI - Convalescent Blood for Treatment of Herpes Zoster Ophthalmicus: Second Report. PMID- 16693211 TI - Keratoconjunctivitis Sicca. PMID- 16693212 TI - A Case of Leptotrichosis Conjunctivae (Parinaud's Conjunctivitis) Without Glandular Involvement. PMID- 16693213 TI - The Site of the Disturbance in Tonic Pupils. PMID- 16693214 TI - The Protein Content of the Reformed Aqueous Humor in Man. PMID- 16693215 TI - Experimental Acute Iridocyclitis in Rabbits Produced by Coliform Bacteria Isolated from the Upper Respiratory Tract of Man. PMID- 16693216 TI - Function and Structure of the Eye. PMID- 16693217 TI - The Histologic Study of an Eye from a Child Suffering with Arachnodactyly. PMID- 16693218 TI - The Significance of Sturm's Interval in Refraction. PMID- 16693219 TI - The Lens in Secondary Glaucoma. PMID- 16693220 TI - A Rare Form of Glaucoma. PMID- 16693221 TI - Intradiem Tension Studies in Chronic Simple Glaucoma. PMID- 16693222 TI - Preservation of Convergence with Paralysis of All Lateral Movements in a Case of Intramedullary Tumor of the Pons. PMID- 16693223 TI - Late Developments in a Case of Gyrate Atrophy of the Choroid and Retina (Fuchs). PMID- 16693224 TI - Heparin in Thrombosis of the Central Vein. PMID- 16693225 TI - Variations in the Development and Regression of Bergmeister's Papilla and the Hyaloid Artery. PMID- 16693226 TI - Drusen of the Optic Papilla: A Clinical and Pathologic Study. PMID- 16693227 TI - Presentation of New Instruments: The New Scotometer. PMID- 16693229 TI - Rhinosporidiosis seeberi Infection in the Eye. PMID- 16693228 TI - Fractures of the Orbit: The Demonstration of the Orbit by Planigraphy (Body Section Radiography). PMID- 16693230 TI - The Sterilization of Sharp Instruments by Chemical Solutions. PMID- 16693231 TI - The Eye in Hypertensive Cardiovascular Disease: A Comparative Ophthalmoscopic and Pathologic Study. PMID- 16693232 TI - The Insertions of the Levator Palpebrae Muscle. PMID- 16693233 TI - Ocular Lymphogranuloma Venereum. PMID- 16693235 TI - Keloid of the Cornea. PMID- 16693234 TI - Retinal Hemorrhages in the Newborn. PMID- 16693236 TI - Equatorial Scleral Staphyloma and Retinal Detachment Cured by Excision. PMID- 16693237 TI - Vitamins of the B Group and Their Relation to Ophthalmology. PMID- 16693238 TI - Death Following Operation for Cataract: Report of Three Cases. PMID- 16693239 TI - Removal of the Lens in High Myopia in which Lenticular Opacities Prevent Improvement of Vision by Correcting Lenses. PMID- 16693240 TI - Neuromyoarterial Glomus Tumor in the Eyelid. PMID- 16693241 TI - Psammoma of the Orbit. PMID- 16693243 TI - Cataract Extraction with Iridodialysis. PMID- 16693242 TI - A Scotoma Associated with Menstruation. PMID- 16693244 TI - Procedures in Intracapsular Cataract Extraction: A New Method. PMID- 16693245 TI - Results of Cataract Extraction. PMID- 16693247 TI - A Survey of Records of Glaucoma Patients in Eye Clinics. PMID- 16693246 TI - Rhabdomyosarcoma of the Orbit. PMID- 16693248 TI - Doryl in the Treatment of Glaucoma Simplex. PMID- 16693249 TI - Cyclodiathermy: An Operation for the Treatment of Glaucoma. PMID- 16693251 TI - Moore's Subjective "Lightning Streaks". PMID- 16693250 TI - Ocular Kinetic Righting Reflexes. PMID- 16693252 TI - Treatment of Blepharospasm. PMID- 16693253 TI - Papilledema Without Increased Intracranial Pressure. PMID- 16693254 TI - Ring Ulcer of the Cornea. PMID- 16693256 TI - Cultural Studies on Patients with Uveitis and Other Eye Diseases. PMID- 16693255 TI - Ocular Pemphigus and its Relation to Pemphigus of the Skin and Mucous Membranes. PMID- 16693257 TI - Gonococcal Conjunctivitis: A Comparison of Sulfanilamide, Sulfapyridine, and Sulfathiazole in the Treatment of 120 Cases. PMID- 16693259 TI - Presentation of New Instruments, Etc. PMID- 16693258 TI - New Aspects of Ocular Micro-organisms Revealed by the Electron Microscope. PMID- 16693260 TI - Familial Degeneration of the Macula Lutea: A Review of the Literature, with a Report of Eight Additional Cases. PMID- 16693261 TI - An Evaluation of Enlargement of the Blind Spot of Mariotte in Brain Trauma in Adults. PMID- 16693262 TI - A Study of Dinitrophenol and Its Relation to Cataract Formation. PMID- 16693263 TI - Concerning Certain Phases of the Development of the Vitreous: A Demonstration of Lantern Slides. PMID- 16693264 TI - Reconstruction of the Eyelids. PMID- 16693265 TI - The Surgical Removal of Corneal Scars. PMID- 16693266 TI - The Treatment of Experimental Bacillus Pyocyaneus Ulcer of the Cornea with Sulfapyridine. PMID- 16693267 TI - Studies on Pigmentary Degeneration of the Retina: The Role of the Melanophore Hormone of the Pituitary Gland in Pigmentary Degeneration of the Retina. PMID- 16693269 TI - Lesions in the Lens Caused by Purulent Corneal Ulcers. PMID- 16693268 TI - Roentgenography of Exophthalmos, with Notes on the Roentgen Ray in Ophthalmology. PMID- 16693270 TI - Alterations in the Capsular Epithelium in Immature Cataract. PMID- 16693272 TI - Retinal Periphlebitis in the Course of Acute Exudative Choroiditis. PMID- 16693271 TI - The Operative Treatment of Congenital Subluxation of the Lens. PMID- 16693274 TI - Surgery of the Inferior Oblique at or Near the Insertion. PMID- 16693273 TI - Wound Rupture After Cataract Extraction. PMID- 16693275 TI - The Effect of Mydriatics Upon the Intra-ocular Pressure in So-Called Primary Wide Angle Glaucoma. PMID- 16693276 TI - Some Observations on the Results of Densensitization in Tuberculous Iritis. PMID- 16693277 TI - Experimental Studies in Ocular Tuberculosis: VII. Effect of Desensitization with Tuberculin in Experimental Ocular Tuberculosis. PMID- 16693278 TI - Allergic Keratoconjunctivitis. PMID- 16693279 TI - Corneal Vascularization Problems. PMID- 16693280 TI - Local Sulfonamide Therapy of Catarrhal Conjunctivitis. PMID- 16693281 TI - Precancerous Melanosis and Malignant Melanoma of the Conjunctiva and Skin of the Lids. PMID- 16693282 TI - Metastatic Orbital Infection: Report of a Case. PMID- 16693283 TI - Observations on the Physiology of the Aqueous in Completely Iridectomized Eyes. PMID- 16693284 TI - A Colored Reflex from the Anterior Capsule of the Lens Which Occurs in Mercurialism. PMID- 16693285 TI - Fibroblastic Overgrowth of Persistent Tunica Vasculosa Lentis in Infants Born Prematurely: II. Report of Cases-Clinical Aspects. PMID- 16693286 TI - Abscess of the Crystalline Lens. PMID- 16693287 TI - Cataract Complicating Corneal Scars after Perforating Ulcers. PMID- 16693288 TI - Studies of the Fields of Vision in Cases of Rathke Pouch Tumors. PMID- 16693289 TI - Tumor Formation at the Disc. PMID- 16693290 TI - Extrapapillary Coloboma. PMID- 16693291 TI - Presentation of New Instruments: A Confirmation Test for Refraction. PMID- 16693292 TI - Progressive Myopia: A Possible Etiologic Factor. PMID- 16693293 TI - Endophthalmitis with Secondary Glaucoma Accompanying Absorption of the Crystalline Lens. PMID- 16693294 TI - Further Observations on Autofundoscopy (Auto-Ophthalmoscopy of Eber; Purkinje Figure of Walker). PMID- 16693295 TI - Herpes Zoster Ophthalmicus: Report of Cases and a Review of the Literature. PMID- 16693296 TI - George Berkeley and "An Essay Towards a New Theory of Vision". PMID- 16693297 TI - The Ocular Pathology of Methyl Alcohol Poisoning. PMID- 16693298 TI - Transscleral Lacrimal Canaliculus Transplants. PMID- 16693299 TI - An Experimental Study of Iridodialysis. PMID- 16693300 TI - The Use of Conjunctival Flaps in Cataract Extraction. PMID- 16693301 TI - A New Cross-Cylinder Test for Ascertaining the Astigmatic Axis, Without the Use of Test Types. PMID- 16693302 TI - Objective Strabismometry in Young Children. PMID- 16693303 TI - Correction of Low Astigmia. PMID- 16693305 TI - A Reply to Criticisms of Aniseikonia. PMID- 16693304 TI - Practical Importance of Aniseikonia. PMID- 16693306 TI - Epidemic Keratoconjunctivitis from a Subjective Viewpoint. PMID- 16693307 TI - Relationship Between the Bacteriology of the Conjunctiva and Nasal Mucosa, with Especial Reference to Certain Extra-Ocular Inflammatory Diseases. PMID- 16693308 TI - The American Board of Ophthalmology Learns About Written Examinations. PMID- 16693310 TI - A Report of Cases of Metastasis of Malignant Tumors to the Eye. PMID- 16693309 TI - The Distribution of Certain Oxidative Enzymes in the Ciliary Body. PMID- 16693311 TI - A Problem of Split Macula-Visual Field Study. PMID- 16693313 TI - So-Called Primary Retinal Tuberculosis. PMID- 16693312 TI - Retinal Detachment and Trauma. PMID- 16693314 TI - Boeck's Sarcoid of Retina-Miliary Form. PMID- 16693316 TI - Keratectomies for the Treatment of Corneal Opacities. PMID- 16693315 TI - Ophthalmoscopic Classification of Hypertensive Diseases. PMID- 16693317 TI - Temporal Arteritis: Case Report with Eye Findings. PMID- 16693318 TI - Pigment Freckles of the Iris (Benign Melanomas): Their Significance in Relation to Malignant Melanoma of the Uvea. PMID- 16693319 TI - Pathology of the Lens in Non-Traumatic Iritis. PMID- 16693320 TI - Less Evident Causes of Lowered Acuity in Senility. PMID- 16693321 TI - Recurring Attacks of Concomitant Exotropia, Each Followed by Transient Esotropia Migraine the Probable Cause. PMID- 16693322 TI - Traumatic Enophthalmos. PMID- 16693323 TI - Recession of the Trochlea for Reducing the Action of the Superior Oblique Muscle. PMID- 16693324 TI - The Choice of the Fixating Eye in Paralytic and Non-Paralytic Strabismus. PMID- 16693325 TI - A Case of Tuberous Sclerosis. PMID- 16693326 TI - Choked Discs and Low Intrathecal Pressure Occurring in Brain Tumor. PMID- 16693327 TI - Inclusion Blennorrhea. PMID- 16693328 TI - Epithelial Tumors of the Iris. PMID- 16693330 TI - A New Treatment of Foreign Bodies of the Cornea. PMID- 16693329 TI - The Etiology and Treatment of Tobacco-Alcohol Amblyopia. PMID- 16693331 TI - Treatment of Ocular Tuberculosis. PMID- 16693332 TI - Patron Saints of the Eyes. PMID- 16693333 TI - Lipaemia Retinalis in the Non-Diabetic. PMID- 16693334 TI - The Association Between Retinopathies and Encephalopathies in the Common Cardiovasculorenal Affections. PMID- 16693335 TI - Myasthenia Gravis and Its Ocular Signs: A Review. PMID- 16693336 TI - Further Experiences with a System of Intracapsular Cataract Extraction. PMID- 16693337 TI - Notes on Keratoconjunctivitis Sicca. PMID- 16693338 TI - Removal of the Ruptured Capsule in Cataract Operations. PMID- 16693339 TI - Complicated Cataract in Spontaneous Detachment of the Retina. PMID- 16693341 TI - Observations on Penicillin Therapy in Ophthalmology. PMID- 16693340 TI - Toxoplasmosis (Report of Eye Findings in Infant Twins). PMID- 16693342 TI - Deep-Chamber Glaucoma Due to the Formation of a Cuticular Product in the Filtration Angle. PMID- 16693343 TI - Errors in Diagnosis of Intra-Ocular Tumors, Suspected or Real. PMID- 16693344 TI - Bilateral Metastatic Carcinoma of the Choroid with X-Ray Therapy to One Eye: Report of a Case. PMID- 16693345 TI - Spontaneous Retinal Reattachment. PMID- 16693346 TI - Binocular and Red-Free Ophthalmoscopy. PMID- 16693347 TI - Analysis of Aniseikonia Cases. PMID- 16693348 TI - Johannes Muller-A Sketch of His Life and Ophthalmologic Works. PMID- 16693349 TI - Visual Symptoms Caused by Digitalis. PMID- 16693350 TI - Albinism with Co-Existing Anomalies of the Central Nervous System. PMID- 16693351 TI - Amyloid Disease of the Conjunctiva. PMID- 16693352 TI - Diabetic Iridopathy. PMID- 16693354 TI - Congenital Glaucoma and Cataract, Bilateral; Goniotomy and Needling: Case Report. PMID- 16693353 TI - Angioid Streaks in the Deep Layers of the Retina. PMID- 16693355 TI - Further Study of the Glaucoma Due to Peripheral Anterior Synechiae Following Delayed Restoration of the Anterior Chamber After Cataract Operation. PMID- 16693356 TI - The Combined Intracranial and Orbital Operation for Bilateral Retinoblastoma. PMID- 16693357 TI - The Pathogenesis of Intermittent Exophthalmos. PMID- 16693358 TI - Recurrent Erosion of the Cornea. PMID- 16693359 TI - Some Factors Concerned in the Mitotic and Wound-Healing Activities of the Corneal Epithelium. PMID- 16693360 TI - Retrolental Fibroplasia in the Premature Infant: V. Further Studies on Fibroplastic Overgrowth of the Persistent Tunica Vasculosa Lentis. PMID- 16693361 TI - Exhibition of New Instruments: Trial Frame for Young Children. PMID- 16693362 TI - An Analysis of the Changes in Refraction (Based on a Study of the Case Histories of Dr. Edward Jackson). PMID- 16693363 TI - Resection of the Levator Palpebrae for Ptosis with Anatomic Studies. PMID- 16693364 TI - Rubeosis Iridis Diabetica. PMID- 16693365 TI - Some Observations on Divergent Strabismus with Anomalous Retinal Correspondence. PMID- 16693366 TI - Iodide Therapy for Senile Macular Degeneration. PMID- 16693367 TI - An Evaluation of Visual Acuity Symbols. PMID- 16693368 TI - Direct Utilization of the Eye as a Camera. PMID- 16693370 TI - Sympathetic Ophthalmia Following Sub-Conjunctival Rupture of the Eyeball. PMID- 16693369 TI - Factors Affecting Postoperative Hemorrhage in Cataract Extractions. PMID- 16693371 TI - Diathermy Operation for Retinal Detachment; Comparative Results of Different Types of Electrodes. PMID- 16693372 TI - Technical Uses of Air in Ophthalmology. PMID- 16693373 TI - The Relationship of Rubella in the Mother to Congenital Cataracts in the Child. PMID- 16693374 TI - Observations on the Vossius Ring. PMID- 16693375 TI - Persistence and Hyperplasia of the Primary Vitreous: Tunica Vasculosa Lentis or Retrolental Fibroplasia. PMID- 16693376 TI - The Exposing and Fixing of the Eye in the Early Days of Cataract Extraction. PMID- 16693377 TI - Concerning Ocular Imagery. PMID- 16693378 TI - Paralysis of Ocular Elevation with and without Ptosis. PMID- 16693379 TI - A Comparative Study of Sutures Employed in Surgery of the Extra-Ocular Muscles. PMID- 16693380 TI - Some Physiologic Factors in Differential Diagnosis of Superior Rectus and Superior Oblique Paralysis. PMID- 16693381 TI - Retinal Vessel Proliferation in Diabetes. PMID- 16693382 TI - Cultivation of Human Tumor in the Anterior Chamber of the Guinea Pig's Eye. PMID- 16693383 TI - Marginal Myotomy-An Analysis of Twenty-two Cases. PMID- 16693384 TI - Penicillin in Gonococcic Conjunctivitis-Its Use in 30 Cases, Compared with the Sulfonamides in 173 Cases. PMID- 16693385 TI - Plastic Eye Implant. PMID- 16693386 TI - A Device for Group Demonstration of Astigmatism Tests. PMID- 16693387 TI - Exophthalmos Caused by Eosinophilic Granuloma of Bone. PMID- 16693388 TI - Indications and Contraindications for Keratoplasty and Keratectomies. PMID- 16693389 TI - Cataract in Intra-Ocular Tumors. PMID- 16693390 TI - Intra-Ocular Manifestations of Acute Disseminated Lupus Erythematosus: Report of a Case. PMID- 16693391 TI - An Accurate Method for Centering a Front Stop (During Refraction under Cycloplegia). PMID- 16693392 TI - Exhibition of New Instruments: A Modification of the Souter Tonometer. PMID- 16693393 TI - Sensorial Retinal Relationship in Concomitant Strabismus. PMID- 16693394 TI - An Ophthalmologist in the ETO. PMID- 16693395 TI - Unequal Tension as a Sign in Early Glaucoma. PMID- 16693396 TI - The Vascular Basis of Allergy of the Eye and Its Adnexa. PMID- 16693397 TI - Retinal Arterioles in the Hypertensions of Pregnancy. PMID- 16693398 TI - The Field of Vision in Chronic Glaucoma: A Comparison of Full with Reduced Illumination. PMID- 16693400 TI - Presentation of Gavel Block. PMID- 16693399 TI - Penicillin Therapy in Ophthalmology: Concentration of Penicillin in the Aqueous Humor Following Parenteral Injection. PMID- 16693401 TI - Early Ocular Manifestations in the Laurence-Moon-Biedl Syndrome. PMID- 16693402 TI - Dust-Borne Infection in Ophthalmic Surgery. PMID- 16693403 TI - Preliminary Report of Corneal Section with Long Bevel and Conjunctival Flap for Cataract Extraction. PMID- 16693405 TI - Pigmentation of the Palbebral Conjunctiva Resulting from Mascara. PMID- 16693404 TI - The Value of Corneal Peeling or Corneal Resection in Ophthalmology. PMID- 16693406 TI - The Diagnosis of Early Glaucoma. PMID- 16693407 TI - Contribution to the Chemistry of the Aqueous Humor in the Glaucomas. PMID- 16693409 TI - Visual Disturbances Due to Digitalis. PMID- 16693408 TI - Ophthalmology During the War and in the Future. PMID- 16693410 TI - Ocular Changes in Acute Disseminated Lupus Erythematosus: Report of a Case with Microscopic Findings. PMID- 16693411 TI - Lipemia Retinalis: Report of a Case. PMID- 16693413 TI - Pseudotumors of the Macula. PMID- 16693412 TI - Dicumarol and Rutin in Vascular Disorders. PMID- 16693414 TI - Experimental Studies in Retrobulbar Neuritis. PMID- 16693415 TI - The Duction Test-An Evaluation of the Various Technics. PMID- 16693416 TI - Some Observations on the Surgical Treatment of the Extra-ocular Muscles. PMID- 16693417 TI - Medical Assistance at Professional Level. PMID- 16693418 TI - The Dissatisfied Refraction Patient. PMID- 16693419 TI - Evisceration With Intrascleral Implant: A New Technique. PMID- 16693420 TI - Tenotomy of the Superior Oblique for Hypertropia: Preliminary Report. PMID- 16693421 TI - Exhibition of New Instruments: Presenting a Combination Red Filter and Occluder with Several Uses. PMID- 16693422 TI - Observation on Corneal and Conjunctival Pigmentation Occurring Among Workers Engaged in the Manufacture of Hydroquinone. PMID- 16693423 TI - An Analysis and Interpretation of Anomalous Localization and Allied Phenomena Associated With Strabismus. PMID- 16693424 TI - A Study of the Vitreous Pressure in Cataract Surgery. PMID- 16693425 TI - The Autonomic Nervous System in Ocular Diseases. PMID- 16693426 TI - Cataract Complicating Glaucoma. PMID- 16693427 TI - The Surgical Correction of Paresis of the Superior Oblique. PMID- 16693428 TI - The Development of the Ectodermal Framework of the Optic Nerve, With Especial Reference to the Glial Lamina Cribrosa. PMID- 16693429 TI - Mercury in the Lens (Hydrargyrosis Lentis). PMID- 16693430 TI - An Evaluation of Methods and Results of Cataract Operations, as Performed in Private and Clinic Practice at Stanford Hospital. PMID- 16693431 TI - Cephalosporium Keratitis. PMID- 16693432 TI - Corneal Wound Healing II: Variations in Adhesive Power of Fibrin. In Vitro Studies. PMID- 16693433 TI - Antitoxin Treatment of Staphylococcic Corneal Ulceration. PMID- 16693435 TI - The Extraction of Cataract in the Presence of Fluid Vitreous. PMID- 16693434 TI - The Ocular Syndrome of Cataract and Papilledema in the Manifest Form of Parathyroid Deficiency. PMID- 16693436 TI - Further Experiences With Integrated Eyes and Vitallium Implants. PMID- 16693437 TI - Studies in Experimental Ocular Tuberculosis. The Effect of Promin and Promizole in Experimental Ocular Tuberculosis in the Immune-Allergic Rabbit. PMID- 16693438 TI - Diathermy Treatment of Angioma of the Retina. PMID- 16693439 TI - Successful Diathermy Treatment in a Case of Recurring Retinal Hemorrhages and Retinitis Proliferans. PMID- 16693441 TI - Clinical Aniseikonia. PMID- 16693440 TI - The Ocular Fundus in Pheochromocytoma of the Adrenal Gland. Report of 3 Cases. PMID- 16693442 TI - The Hyperophthalmopathy of Graves' Disease. PMID- 16693443 TI - The Use of Buccal Mucosa in Restoration of the Orbital Socket. PMID- 16693444 TI - Mikulicz's Disease and Syndrome. PMID- 16693445 TI - The Diagnosis of Retinoblastoma. PMID- 16693446 TI - Agnostic Alexia Without Agraphia, Following Trauma. Report of a Case. PMID- 16693447 TI - The Significance of the Base Pressure in Primary Glaucoma. PMID- 16693448 TI - Alteration in Plasma Proteins in Various Types of Cataract. PMID- 16693449 TI - Diffuse Malignant Melanoma of the Iris. Report of Two Cases. PMID- 16693450 TI - The Clinical Course of the Eye Complications of Arachnodactylia. PMID- 16693451 TI - Some Problems in the Calibration of Tonometers. PMID- 16693452 TI - Chloroma: Report of a Case and a Review of the Literature. PMID- 16693454 TI - Presentation of Instruments-Instruments for Cataract Surgery. PMID- 16693453 TI - Pupillometry: A Comment. PMID- 16693455 TI - Experimental Studies in Inclusion Blennorrhea. Isolation of a Virus. PMID- 16693456 TI - Ocular Onchocerciasis in Guatemala. An Investigation of 1,215 Natives Infected With Onchocerca Volvulus. PMID- 16693457 TI - Fractures of the Orbital Floor. PMID- 16693458 TI - Myopia and Hypothyroidism. PMID- 16693459 TI - Glaucoma Following Intracapsular Cataract Extraction. PMID- 16693460 TI - The Ciliary Muscle and Insulin Hypoglycemia. PMID- 16693461 TI - Retinopathy of Multiple Sclerosis. PMID- 16693462 TI - Classification of Retinal Lesions in the Presence of Vascular Hypertension: Report submitted to the American Ophthalmological Society by the committee on Classification of Hypertensive Disease of the Retina. PMID- 16693463 TI - Controversial Points in Ocular Penicillin Therapy. PMID- 16693464 TI - Ocular Changes in Scrub Typhus. A Study of 451 Patients. PMID- 16693465 TI - Discoloration of the Eyelids from Prolonged Use of Ointments Containing Mercury. PMID- 16693466 TI - Delayed Mustard Gas Keratitis (Dichlorodiethyl Sulfide). A Report of Two Cases. PMID- 16693467 TI - Granular Cell Myoblastoma of the Orbit. PMID- 16693468 TI - Keratoplasty for the Treatment of Keratoconus. PMID- 16693469 TI - The Surgical Anatomy of the Superior Oblique Muscle. PMID- 16693470 TI - A Simple Appositional Suture in Operations for Cataract. PMID- 16693471 TI - Therapy of Some Ocular Inflammations Based on Immunologic Principles. PMID- 16693472 TI - Primary Malignant Melanoma of the Optic Disc. Report of a Case. PMID- 16693473 TI - Dermoids and Epidermoids of the Orbit. PMID- 16693475 TI - Some Secondary Subjective Effects Produced by Prisms. PMID- 16693474 TI - The Dupuy-Dutemps Dacryocystorhinostomy. PMID- 16693476 TI - "Myopia Cures". PMID- 16693477 TI - The Mechanics of Intracapsular Cataract Extraction. PMID- 16693478 TI - Retinoblastoma in the Adult. PMID- 16693479 TI - Familial Central and Peripapillary Choroidal Sclerosis Associated with Familial Pseudoxanthoma Elasticum. PMID- 16693480 TI - Surgical Procedures in Retinal Vascular Disease. PMID- 16693481 TI - The Epidemiology of Epidemic Keratoconjunctivitis. PMID- 16693482 TI - Ocular Lymphomas. PMID- 16693483 TI - Perineuritis Optica. PMID- 16693484 TI - Aerobic Glycolysis of Corneal Tissue. PMID- 16693485 TI - A Study of the Effect of Retrobulbar Anesthesia on the Intra-ocular Tension and the Vitreous Pressure. PMID- 16693487 TI - Cataract and Tetany Produced by Parathyroid Deficiency During Pregnancy, Lactation and Menstruation. PMID- 16693486 TI - Choice of a Miotic Agent Following Retrobulbar Anesthesia. PMID- 16693488 TI - A New Instrument: The Harrington Tonometer. PMID- 16693489 TI - Vascularization of the Cornea. Its Experimental Induction by Small Lesions and a New Theory of Its Pathogenesis. PMID- 16693490 TI - Enucleation and Allied Procedures: A Review, and Description of a New Operation. PMID- 16693491 TI - Amblyopia Ex Anopsia. Observations on Retinal Inhibition, Scotoma, Projection, Light Difference Discrimination and Visual Acuity. PMID- 16693492 TI - Evaluation of Night Vision. PMID- 16693493 TI - Scleral Staphyloma and Retinal Detachment. PMID- 16693494 TI - The Surgical Treatment of Recurrent Pterygia. PMID- 16693495 TI - Direct Surgery of Underacting Oblique Muscles. PMID- 16693496 TI - Ocular Effects of the Choline Esters. PMID- 16693497 TI - Herniation of the Anterior Hyaloid Membrane Following Uncomplicated Intracapsular Cataract Extraction. PMID- 16693498 TI - The Effect of Dicumarol on the Visual Fields in Glaucoma. A Preliminary Report. PMID- 16693499 TI - The President's Badge. PMID- 16693500 TI - The Mode of Action of Iris Inclusion Operations. PMID- 16693501 TI - A Gonioscopic Study of Fistulizing Operations. PMID- 16693503 TI - Emmetropia. PMID- 16693502 TI - Malignant Glaucoma. PMID- 16693504 TI - Ophthalmia Nodosa. PMID- 16693506 TI - Fundus Changes in Juvenile Diabetics. PMID- 16693505 TI - The Dynamics of Phakoerisis. PMID- 16693507 TI - Diabetic Retinopathy in Pregnancy. PMID- 16693508 TI - Crystalline Dystrophy of the Cornea. PMID- 16693509 TI - Experimental Studies on Early Lens Changes after X-Ray Irradiation. PMID- 16693510 TI - The Early Years of Helmholtz: In Commemoration of the Centenary of the Invention of the Ophthalmoscope. PMID- 16693511 TI - Lint in the Anterior Chamber Following Intraocular Surgery. PMID- 16693512 TI - Clinical and Experimental Observation on the Use of ACTH and Cortisone in Ocular Inflammatory Disease. PMID- 16693513 TI - Total Penetrating Keratoplasty: Preliminary Report. PMID- 16693514 TI - Relationship of the Endocrines to Cataract. PMID- 16693515 TI - New Instruments: A New Needle Holder. PMID- 16693516 TI - The Relationship of Pseudoptosis to Muscle Tropias and the Palpebral Aperture: With Special Reference to the Surgical Correction of Pseudoptosis. PMID- 16693518 TI - The First Ten Years of Ophthalmoscopy, 1851-1861. PMID- 16693517 TI - Correlation of Surgical Treatment of Essential Hypertension with Blood Pressure Response Tests and Retinal Changes. PMID- 16693519 TI - Psychoneurotic Reaction Patterns in Ophthalmology. PMID- 16693521 TI - Clinical Applications of Night Vision Tests. PMID- 16693522 TI - The Action of the Extraocular Muscles: A Method of Vector-Analysis with Computations. PMID- 16693523 TI - Congenital and Familial Endothelial Defects. PMID- 16693524 TI - Ocular Manifestations of Multiple Sclerosis and Relationship to Retrobulbar Neuritis. PMID- 16693525 TI - Hydrophthalmia in Identical Twins. PMID- 16693526 TI - Ocular Changes Following Aortic Pulmonary Anastomosis for Congenital Heart Disease. PMID- 16693527 TI - Pseudoglioma: A Clinico-Pathologic Study of Fifteen Cases. PMID- 16693530 TI - An Analysis of Methods for Measuring Diplopia Fields and an Introduction of a Device for Measurements of This Sort. PMID- 16693529 TI - Ophthalmologic Survey of Atomic Bomb Survivors in Japan, 1949. PMID- 16693531 TI - Diabetic Retinopathy in Young Persons. PMID- 16693532 TI - Ocular Hypertension Induced by Air in the Anterior Chamber. PMID- 16693533 TI - A Report on a Series of 401 Cataract Extractions with Special Reference to the Keratome-Scissors Incision. PMID- 16693534 TI - The Results of Cataract Extraction in Cases of Complicated Cataracts. PMID- 16693535 TI - Tumors of the Iris Muscle. PMID- 16693536 TI - Embryonic Intra-Epithelial Cyst of the Ciliary Processes. PMID- 16693537 TI - Terramycin in Ocular Infections. PMID- 16693538 TI - Retrolental Fibroplasia in Premature Infants, III: The Pathology of the Disease. PMID- 16693539 TI - Scleromalacia: Clinical and Pathologic Study of a Case with Consideration of Differential Diagnosis, Relationship of Collagen Disease, and Effect of Acth and Cortisone Therapy. PMID- 16693540 TI - Experimental and Clinical Observations of the Effects of Cortisone in Corneal Lesions. PMID- 16693541 TI - Choriopathy. PMID- 16693542 TI - Superficial Punctate Keratitis: The Isolation of a Virus. PMID- 16693543 TI - A Simplified Blaskovics Operation for Blepharoptosis: Results in 91 Operations. PMID- 16693544 TI - Diagnostic and Prognostic Implications of Vitreous Opacities. PMID- 16693546 TI - Polyarteritis Nodosa of the Eye. PMID- 16693545 TI - Optic Atrophy in Infancy, Childhood, and Adolescence: A Survey of 81 Cases. PMID- 16693548 TI - Visual Field Changes Due to Tumors of the Choroid. PMID- 16693547 TI - Further Efforts to Influence X-Ray Cataract by Chemical Agents. PMID- 16693549 TI - Changes in the Eye and Orbit Following Carotid Ligation. PMID- 16693550 TI - Fundus Changes in Leukemia. PMID- 16693551 TI - The Association of Uveal Nevi with Skin Nevi. PMID- 16693552 TI - A Corneal Contact Prism and a Corneal Contact Lens for Slit Lamp Biomicroscopic Examination of the Peripheral and Central Fundus. PMID- 16693554 TI - The Use of the Sanborn Electromanometer in the Study of Pharmacological Effects upon the Intraocular Pressure. PMID- 16693553 TI - Ocular Aspects of Mycotic Infection. PMID- 16693555 TI - An Experimental Study on the Evaluation of Hydrosulphosol in the Treatment of Ocular Injuries Due to Chemical Burns. PMID- 16693556 TI - Suprasellar Meningioma: Report of an Atypical Case. PMID- 16693557 TI - The Relationship of Lenticular Changes to Mongolism. PMID- 16693558 TI - Autonomic Drugs and Their Influence on Chorioidal Vessel Caliber. PMID- 16693559 TI - Early Wounds in and about the Orbit. PMID- 16693560 TI - Pieces of Retina in the Vitreous. PMID- 16693561 TI - Complications of Cataract Surgery. PMID- 16693562 TI - Necrology: Gideon McDonald Van Poole, M.D. PMID- 16693563 TI - Experiences with Light Coagulation of Fundus Lesions. PMID- 16693564 TI - The Effect of Paralysis of the Cervical Sympathetics on the Electromyogram of Extraocular Muscles in Man. PMID- 16693565 TI - Long-term Preservation of Donor Tissue for Corneal Grafting: Correlation of Results from Tissue Cultures with Those from Experimental Graftings. PMID- 16693566 TI - Retinal Dehydrogenases. PMID- 16693567 TI - Ocular Complications of Transsphenoidal Yttrium 90 Hypophysectomy. PMID- 16693568 TI - Hemangioma of the Choroid. PMID- 16693569 TI - Expulsive Subchoroidal Hemorrhage associated with Cataract Extraction: Report of a Case with Survival of the Eye. PMID- 16693570 TI - Cinematography of the Retinal Vessels. PMID- 16693571 TI - Experience with the Jose Barraquer Method of Extracting a Dislocated Lens. PMID- 16693572 TI - Completely Dislocated Hypermature Cataract and Glaucoma. PMID- 16693573 TI - Chymotrypsin: An Aid to Intracapsular Cataract Extraction. PMID- 16693574 TI - The Ocular Features of a Case of Acute Reticuloendotheliosis (Letterer-Siwe Type). PMID- 16693575 TI - Severe Exophthalmos Associated with Malignant Lymphoma. PMID- 16693576 TI - The Probable Role of Benign Histoplasmosis in the Etiology of Granulomatous Uveitis. PMID- 16693577 TI - The Use of Amphotericin B in Selected Cases of Chorioretinitis. PMID- 16693578 TI - Modified Electro-keratotome. PMID- 16693579 TI - New Handles for Razor Blade Knives. PMID- 16693580 TI - Panum's Areas and Some Other Prevailing Misconceptions concerning Binocular Vision. PMID- 16693581 TI - Description of Recession Forceps. PMID- 16693582 TI - Aseptic Technique in Ophthalmology. PMID- 16693584 TI - Ophthalmoscopically Visible Small Spots in the Ocular Fundus. PMID- 16693585 TI - Hereditary Dyskeratosis of the Perilimbal Conjunctiva. PMID- 16693583 TI - Elevated Intravascular Pressure as an Etiologic Mechanism in the Production of Eye Injuries. PMID- 16693586 TI - Pigmentation of the Lens: A Consideration of Endogenous Diffuse and Granular Types. PMID- 16693587 TI - Ocular Manifestations of Thyroid Disease: Current Concepts. PMID- 16693588 TI - Lymphangiomas of the Ocular Adnexa: An Analysis of 62 Cases. PMID- 16693589 TI - The Effects of Anticholinesterase Agents on the Blood Cholinesterases Levels of Normal and Glaucoma Subjects. PMID- 16693590 TI - Vertically Incomitant Horizontal Strabismus: The So-called "A" and "V" Syndromes. PMID- 16693591 TI - The Use of Absorbable Sutures in Surgery of the Cornea. PMID- 16693592 TI - Pathogenesis of Paralysis of the Third Cranial Nerve. PMID- 16693593 TI - Intrascleral Drainage Channels of the Normal Rabbit Eye. PMID- 16693594 TI - Lipid-Proteinosis (Urbach-Wiethe) Involving the Lids. PMID- 16693595 TI - School Vision Testing. PMID- 16693596 TI - Sympathetic Ophthalmia: A Case Treated by Prolonged Use of Corticotropin and Adrenal Steroids. PMID- 16693597 TI - The Frederick H. Verhoeff Lecture: Angioid Streaks, a Kodachrome exposition. PMID- 16693599 TI - Triflupromazine Hydrochloride (Vesprin(R)) Its Value in Eye Surgery. PMID- 16693598 TI - The Use of Corneas from Frozen Whole Eyes for Emergency Keratoplasty. PMID- 16693600 TI - Improved Cutting Edge for Eye Instruments. PMID- 16693601 TI - Hematomas Under the Retinal Pigment Epithelium. PMID- 16693602 TI - Experimental Embolization of the Retinal Arterioles. PMID- 16693603 TI - A Clinical and Experimental Study of Electric Cataract. PMID- 16693604 TI - The Effect of I.D.U. on Experimental and Clinical Herpes Simplex Infections. PMID- 16693605 TI - Non-Syphilitic Interstitial Keratitis with Vestibulo-Auditory Symptoms-A Case with Fatal Aortitis. PMID- 16693606 TI - Ten Years' Experience with D.F.P. in the Handling of Esotropia. PMID- 16693607 TI - Relaxing Procedures of the Inferior Oblique-A Comparative Study. PMID- 16693608 TI - Spiramycin for Toxoplasmosis. PMID- 16693609 TI - Lamellar Keratoplasty Using Full Thickness Donor Material. PMID- 16693610 TI - The Removal of Tritiated Water from the Vitreous of the Dog. PMID- 16693611 TI - Open-Angle Glaucoma Associated with Vitreous Filling the Anterior Chamber. PMID- 16693612 TI - Scleral Cautery with Iridectomy-An Experimental Study. PMID- 16693613 TI - Iridectomy in Advanced Angle-Closure Glaucoma. PMID- 16693614 TI - Dyskeratotic Epibulbar Tumors. PMID- 16693615 TI - Sympathetic Ophthalmia after Evisceration. PMID- 16693616 TI - Further Studies on the Binding of Exophthalmos-producing Substance by Normal Human Serum. PMID- 16693617 TI - Glioma (Astrocytoma) of the Optic Nerve, with Reports of Three Cases, Treated Surgically. PMID- 16693618 TI - Hereditary Vitelline Macular Degeneration: A Clinical and Functional Evaluation of a New Pedigree with Variable Expressivity and Dominant Inheritance. PMID- 16693619 TI - Reconstruction of the Orbit with Polyvinyl Sponge: A Follow-up Report. PMID- 16693620 TI - Anterior Chamber Depth in Aphakia. PMID- 16693621 TI - Chronic Cyclitis. PMID- 16693622 TI - A Modified Ophthalmodynamometer. PMID- 16693623 TI - A Disposable Eight-Day Eye Tray. PMID- 16693624 TI - Metabolism of the Alloxan Diabetic Rat Retina. PMID- 16693625 TI - Ocular Changes Due to Contact Lenses. PMID- 16693626 TI - Triparanol-induced Cataract in Rats. PMID- 16693627 TI - The Role of Cholesterol in the Pathogenesis of Coats' Disease. PMID- 16693628 TI - The Ultrastructure of the Retinal Pigment Epithelium with Observations on the Choriocapillaris and Bruch's Membrane. PMID- 16693629 TI - A Correlative Study of Orbital Vascular and Intracranial Pressure in the Rhesus Monkey. PMID- 16693630 TI - Striate Melanokeratosis in Negroes. PMID- 16693631 TI - Total Hyphema with Special Reference to Treatment of the Experimental Lesion. PMID- 16693632 TI - After-Image Scotometry: A Method of Functional Examination of the Eye. PMID- 16693633 TI - Pathology of Herpes Simplex Kerato-Iritis. PMID- 16693634 TI - Retinal Detachment in Aphakia. PMID- 16693635 TI - External Ophthalmoplegia, Pigmentary Degeneration of the Retina, and Cardiomyopathy: A Newly Recognized Syndrome. PMID- 16693636 TI - AOS, : John Green, theand the Reasonable Notation of Visual Acuity Measurements. PMID- 16693637 TI - PURPOSES IN MEDICAL RESEARCH: An Introduction to the Journal of Clinical Investigation. PMID- 16693638 TI - STUDIES ON THE SPECIFIC GRAVITY OF THE URINE. PMID- 16693639 TI - ON THE RELATION BETWEEN CONDUCTIVITY AND CHLORIDES IN THE URINE. PMID- 16693640 TI - BLOOD REACTION AND BLOOD GASES IN PNEUMONIA. PMID- 16693641 TI - A METHOD FOR THE DETERMINATION OF THE AMOUNT OF OXYGEN AND CARBON DIOXIDE IN THE MIXED VENOUS BLOOD OF MAN. PMID- 16693643 TI - THE GASEOUS CONTENT OF THE BLOOD AND THE OUTPUT OF THE HEART IN NORMAL RESTING ADULTS. PMID- 16693644 TI - EVIDENCE THAT DIGITALIS INFLUENCES CONTRACTION OF THE HEART IN MAN. PMID- 16693642 TI - THE RATE OF THE CIRCULATION OF THE BLOOD IN NORMAL RESTING INDIVIDUALS. PMID- 16693645 TI - EXPERIMENTAL STUDIES ON RAPID BREATHING: I. Tachypnea, Independent of Anoxemia, Resulting from Multiple Emboli in the Pulmonary Arterioles and Capillaries. PMID- 16693646 TI - EXPERIMENTAL STUDIES ON RAPID BREATHING: II. Tachypnea, Dependent upon Anoxemia, Resulting from Multiple Emboli in the Larger Branches of the Pulmonary Artery. PMID- 16693647 TI - TRANSIENT AURICULAR FIBRILLATION FOLLOWING DIGITALIS THERAPY, WITH OBSERVATIONS UPON THE REACTION TO ATROPINE. PMID- 16693648 TI - THE LEUCOCYTE CURVE AS AN INDEX OF THE INFECTION IN RHEUMATIC FEVER. PMID- 16693649 TI - THE ACTION OF PITUITARY EXTRACT ON THE HEART OF THE UNANESTHETIZED DOG. PMID- 16693650 TI - THE ACTION OF PITUITARY TARTRATE ON THE HEART OF THE UNANESTHETIZED DOG. PMID- 16693651 TI - CARBOHYDRATE METABOLISM IN NEPHRITIS. PMID- 16693652 TI - STUDIES IN SCARLET FEVER: I. Studies Concerning the Blanching Phenomenon in Scarlet Fever. PMID- 16693653 TI - BLOOD PIGMENTS IN PERNICIOUS ANEMIA. PMID- 16693654 TI - THE OPTICAL ACTIVITY OF GLUCOSE AS INFLUENCED BY NORMAL AND DIABETIC URINE. PMID- 16693655 TI - OBSERVATIONS ON THE USE OF NOVASUROL IN EDEMA DUE TO HEART FAILURE. PMID- 16693656 TI - A STUDY OF THE DIURETIC ACTION OF ACID PRODUCING SALTS. PMID- 16693657 TI - CLINICAL OBSERVATIONS ON THE T WAVE OF THE AURICLE APPEARING IN THE HUMAN ELECTROCARDIOGRAM. PMID- 16693658 TI - THE FACTORS IN THE DEHYDRATION FOLLOWING PYLORIC OBSTRUCTION. PMID- 16693659 TI - THE RESISTANCE OF IMMATURE ERYTHROCYTES TO HEAT. PMID- 16693660 TI - THE PLASMA PROTEINS IN RELATION TO BLOOD HYDRATION: I. In Normal Individuals and in Miscellaneous Conditions. PMID- 16693661 TI - THE PLASMA PROTEINS IN RELATION TO BLOOD HYDRATION: II. In Diabetes Mellitus. PMID- 16693662 TI - IODINE STUDIES: I. The Avidity of the Thyroid Gland for Various Iodine Compounds in Vitro. PMID- 16693663 TI - THE EFFECT OF SOME PATHOLOGICAL CONDITIONS UPON DYSPNEA DURING EXERCISE: I. Artificial Stenosis. PMID- 16693665 TI - A STUDY OF SEROLOGICAL REACTIONS ASSOCIATED WITH EXPERIMENTAL PLETHORA AND PLETHORIC ANEMIA. PMID- 16693664 TI - THE MECHANISM OF DEATH FROM QUINIDINE AND A METHOD OF RESUSCITATION; AN EXPERIMENTAL STUDY. PMID- 16693666 TI - A STUDY OF THE EFFECTS OF PYLORIC OBSTRUCTION IN RABBITS. PMID- 16693667 TI - THE EFFECTS OF CHANGES IN HYDROGEN ION CONCENTRATION ON THE BLOOD FLOW OF MORPHINIZED DOGS. PMID- 16693668 TI - CLINICAL AND EXPERIMENTAL STUDIES ON SODIUM BENZOATE: The Value of the Sodium Benzoate Test of Renal Function, and the Effect of Injury of the Liver on Hippuric Acid Synthesis. PMID- 16693669 TI - STUDIES ON THE MORNING ALKALINE TIDE OF URINE IN NORMAL PERSONS AND IN PATIENTS WITH NEPHRITIS. PMID- 16693670 TI - ORTHOPNOEA AND THE EFFECT OF POSTURE UPON THE RATE OF BLOOD FLOW. PMID- 16693671 TI - THE ELIMINATION OF CARBON MONOXIDE FROM THE BLOOD: A Theoretical and Experimental Study. PMID- 16693672 TI - OBSERVATIONS ON THE EFFECT OF ANOXEMIA ON THE HEART: I. Auriculo-ventricular Conduction. PMID- 16693673 TI - OBSERVATIONS OF THE EFFECT OF ANOXEMIA ON THE HEART: II. Intraventricular Conduction. PMID- 16693674 TI - OBSERVATIONS ON THE EFFECT OF ANOXEMIA ON THE HEART: III. Changes in the Auricles, with Particular Reference to the Relationship Between Anoxemia and Auricular Fibrillation. PMID- 16693675 TI - SIMULTANEOUS RESPIRATORY EXCHANGE AND BLOOD SUGAR TIME CURVES OBTAINED IN DIABETIC AND NON DIABETIC INDIVIDUALS FOLLOWING INGESTION OF GLUCOSE. PMID- 16693676 TI - THE EFFECT OF CAFFEIN SODIO-BENZOATE, THEO-BROMIN SODIO-SALICYLATE, THEOPHYLLIN AND EUPHYLLIN ON THE CORONARY FLOW AND CARDIAC ACTION OF THE RABBIT. PMID- 16693677 TI - TOTAL ACID-BASE EQUILIBRIUM OF PLASMA IN HEALTH AND DISEASE: VI. Studies of Diabetes. PMID- 16693678 TI - TOTAL ACID-BASE EQUILIBRIUM OF PLASMA IN HEALTH AND DISEASE: VII. Factors Causing Acidosis in Chronic Nephritis. A Preliminary Report. PMID- 16693679 TI - OBSERVATIONS ON THE EXTRINSIC NERVOUS CONTROL OF THE AURICLES AND VENTRICLES IN COMPLETE AURICULO-VENTRICULAR BLOCK IN MAN. PMID- 16693680 TI - THE EFFECT OF CHANGES IN REGIME UPON THE URINARY ALKALINE TIDES IN A NORMAL INDIVIDUAL. PMID- 16693681 TI - THE EXCRETION OF AMMONIA AND TITRATABLE ACID IN NEPHRITIS. PMID- 16693682 TI - THE EFFECT OF IODINE BY MOUTH ON THE REACTION TO INTRAVENOUS INJECTIONS OF THYROXIN. PMID- 16693683 TI - THE EFFECT OF A STANDARD EXERCISE UPON THE OXYGEN CONSUMPTION OF NORMAL, OVERWEIGHT, AND CHRONICALLY ILL INDIVIDUALS. PMID- 16693684 TI - EFFECT OF PHENYLHYDRAZINE DERIVATIVES IN THE TREATMENT OF POLYCYTHEMIA. PMID- 16693685 TI - EXPERIMENTAL ANEMIA PRODUCED BY PHENYL-HYDRAZINE DERIVATIVES. PMID- 16693686 TI - STUDIES ON HUMAN CAPILLARIES: I. An Apparatus for Cinematographic Observation of Human Capillaries. PMID- 16693687 TI - STUDIES ON HUMAN CAPILLARIES: II. Observations on the Capillary Circulation in Normal Subjects. PMID- 16693689 TI - JAUNDICE FOLLOWING PULMONARY INFARCTION IN PATIENTS WITH MYOCARDIAL INSUFFICIENCY: I. A Clinical Study. PMID- 16693688 TI - STUDIES ON HUMAN CAPILLARIES: III. Observations in Cases of Auricular Fibrillation. PMID- 16693690 TI - JAUNDICE FOLLOWING PULMONARY INFARCTION IN PATIENTS WITH MYOCARDIAL INSUFFICIENCY: II. An Experimental Study. PMID- 16693691 TI - THE NUMBER OF FORMED ELEMENTS IN THE URINARY SEDIMENT OF NORMAL INDIVIDUALS. PMID- 16693692 TI - THE EFFECT OF SOME PHYSIOLOGICAL VARIABLES ON THE NUMBER OF CASTS, RED BLOOD CELLS AND WHITE BLOOD CELLS AND EPITHELIAL CELLS IN THE URINE OF NORMAL INDIVIDUALS. PMID- 16693693 TI - MEASUREMENTS ON THE SKIN CAPILLARIES IN CASES OF POLYCYTHEMIA VERA AND THE ROLE OF THESE CAPILLARIES IN THE PRODUCTION OF ERYTHROSIS. PMID- 16693694 TI - CARDIAC OUTPUT IN PNEUMONIA IN THE DOG. PMID- 16693695 TI - THE UTILIZATION OF CARBOHYDRATES IN A CASE OF CHRONIC PENTOSURIA. PMID- 16693696 TI - A NEW TYPE OF OXYGEN CHAMBER. PMID- 16693698 TI - DIHYDROXYACETONE STUDIES: I. Its Respiratory and Carbohydrate Metabolism in Normal Men. PMID- 16693697 TI - STUDIES IN BLOOD VOLUME: I. The Blood Volume in Myxedema, with A Comparison of Plasma Volume Changes in Myxedema and Cardiac Edema. PMID- 16693699 TI - DIHYDROXYACETONE STUDIES: II. Its Respiratory and Carbohydrate Metabolism in Diabetes Mellitus. PMID- 16693700 TI - THE EFFECT OF ORALLY ADMINISTERED HYDROCHLORIC ACID UPON THE GASTRIC CONTENTS IN NORMAL INDIVIDUALS AND IN PATIENTS WITH ACHLORHYDRIA. PMID- 16693701 TI - BLOOD SUGAR TIME CURVES. PMID- 16693702 TI - GUANIDINE EXCRETION IN RELATION TO HYPERTENSION. PMID- 16693703 TI - THE EFFECT OF DIGITALIS ON THE CARDIAC OUT-PUT OF DOGS AND ITS BEARING ON THE ACTION OF THE DRUG IN HEART DISEASE. PMID- 16693704 TI - THE SIGNIFICANCE OF ELECTROCARDIOGRAMS OF LOW VOLTAGE. PMID- 16693705 TI - KIDNEY FUNCTION IN PNEUMONIA. PMID- 16693706 TI - AN EXPERIMENTAL STUDY OF THE SYNOVIAL FLUID OF PATIENTS WITH ARTHRITIS AND SYPHILIS. PMID- 16693707 TI - A STUDY OF THE ACTION OF AMMONIUM CHLORID AND ORGANIC MERCURY COMPOUNDS. PMID- 16693708 TI - STUDIES IN SERUM ELECTROLYTES: I. Concentration of Electrolytes and Non electrolytes in the Serum during Lobar Pneumonia. PMID- 16693709 TI - AN EXPERIMENTAL STUDY OF CHRONIC AORTIC REGURGITATION IN DOGS. PMID- 16693710 TI - BENIGN GLYCOSURIA DUE TO DISTURBANCES IN THE BLOOD SUGAR REGULATING MECHANISM. PMID- 16693711 TI - STUDIES ON RED CELL DIAMETER: I. In Health and in Pernicious Anemia. PMID- 16693712 TI - THE EFFECT OF INCREASED HEART RATE DUE TO THE INJECTION OF ATROPINE ON THE OXYGEN SATURATION OF THE ARTERIAL AND VENOUS BLOOD OF PATIENTS WITH HEART DISEASE. PMID- 16693713 TI - INFLUENCE OF PREVIOUS SALT REGIME ON EXCRETIONS OF CHLORINE, SODIUM, AND POTASSIUM DURING THE CHLORIDE CONCENTRATION TEST OF DE WESSELOW. PMID- 16693714 TI - THE SURFACE TENSION OF THE BLOOD SERUM IN NEPHRITIS. PMID- 16693715 TI - THE PHYSIOLOGICAL RESPONSE OF THE CIRCULATORY SYSTEM TO EXPERIMENTAL ALTERATIONS: III. The Effect of Aortic and Pulmonic Stenoses. PMID- 16693716 TI - THE ASSOCIATION OF GENERALIZED ARTERIOLAR SCLEROSIS WITH HIGH BLOOD PRESSURE AND CARDIAC HYPERTROPHY IN CHRONIC NEPHRITIS. PMID- 16693717 TI - THE EFFECT OF BREATHING OXYGEN-ENRICHED AIR DURING EXERCISE UPON PULMONARY VENTILATION AND UPON THE LACTIC ACID CONTENT OF BLOOD AND URINE. PMID- 16693718 TI - CALORIMETRIC STUDIES OF THE EXTREMITIES: I. Theory and Practice of Methods Applicable to Such Investigations. PMID- 16693720 TI - CALORIMETRIC STUDIES OF THE EXTREMITIES: III. Clinical Data on Normal and Pathologic Subjects with Localized Vascular Disease. PMID- 16693719 TI - CALORIMETRIC STUDIES OF THE EXTREMITIES: II. Experimental Apparatus and Procedures. PMID- 16693721 TI - STUDIES IN SCARLET FEVER: I. The Amount of Scarlatinal Toxin in the Blood of Patients with Scarlet Fever. PMID- 16693722 TI - STUDIES IN SCARLET FEVER: II. The Relation of the Specific Toxemia of Scarlet Fever to the Course of the Disease. PMID- 16693723 TI - STUDIES IN SCARLET FEVER: III. Infections with Streptococcus Scarlatinae in Persons with Scarlatinal Antitoxic Immunity. PMID- 16693724 TI - STUDIES IN SCARLET FEVER: IV. Post-scarlatinal Immunity in Patients Treated with Antitoxin. PMID- 16693726 TI - THE EFFECT OF TACHYCARDIA ON THE BLOOD FLOW IN DOGS: II. The Effect of Rapid Regular Rhythm. PMID- 16693725 TI - THE EFFECT OF TACHYCARDIA ON THE BLOOD FLOW IN DOGS: I. The Effect of Rapid Irregular Rhythms as Seen in Auricular Fibrillation. PMID- 16693727 TI - EPIDEMIC PREVALENCE IN THE LIGHT OF EXPERIMENTAL FINDINGS. PMID- 16693728 TI - A TECHNIQUE FOR MEASURING X-RAY PHOTOGRAPHS OF THE CARDIAC AREAS OF DOGS. PMID- 16693729 TI - THE EFFECT OF REGULAR AND IRREGULAR TACHYCARDIAS ON THE SIZE OF THE HEART. PMID- 16693730 TI - THE PLASMA PROTEINS IN RELATION TO BLOOD HYDRATION: III. The Plasma Proteins in Malnutrition. PMID- 16693731 TI - TOTAL ACID-BASE EQUILIBRIUM OF PLASMA IN HEALTH AND DISEASE: VIII. Bicarbonate and Chloride in the Serum of Patients with Heart Failure. PMID- 16693732 TI - TOTAL ACID-BASE EQUILIBRIUM OF PLASMA IN HEALTH AND DISEASE: IX. High Serum Bicarbonate in Heart Failure. Asphyctic Anoxemia. PMID- 16693733 TI - DIGITALIS AND DIURETICS IN HEART FAILURE WITH REGULAR RHYTHM, WITH ESPECIAL REFERENCE TO THE IMPORTANCE OF ETIOLOGIC CLASSIFICATION OF HEART DISEASE. PMID- 16693734 TI - STUDIES ON RED BLOOD CELL DIAMETER : II. In Pernicious Anemia, Before and During Marked Remission, and in Myelogenous Leukemia. PMID- 16693736 TI - EPINEPHRIN REACTION IN OBESITY. PMID- 16693735 TI - THE MECHANISM OF THE ACTION OF THE HYDROGEN ION UPON THE CARDIAC RHYTHM. PMID- 16693738 TI - DEPENDENCE OF THE FORM OF THE ELECTRO-CARDIOGRAM UPON THE SITE OF MECHANICAL STIMULATION OF THE HUMAN VENTRICLES. PMID- 16693737 TI - EDEMA: II. The Effectiveness of Ultrafiltration for Quantitatively Determining the "Free Water" Content of Blood Plasma, and for Estimating Physical-Chemical Changes of the Plasma Proteins in Edema. PMID- 16693739 TI - VARIATIONS IN BLOOD FLOW WITH CHANGES IN POSITION IN NORMAL AND PATHOLOGIC SUBJECTS. PMID- 16693741 TI - STUDIES ON THE VELOCITY OF BLOOD FLOW: I. The Method Utilized. PMID- 16693740 TI - A COMPARATIVE STUDY OF THE ROTATORY AND REDUCING PROPERTIES OF ULTRAFILTRATES FROM BLOOD PLASMA. PMID- 16693742 TI - OXY-HEMOGLOBIN DISSOCIATION CURVES OF WHOLE BLOOD IN ANEMIA. PMID- 16693743 TI - THE RELATION BETWEEN THE BLOOD UREA CONCENTRATION AND THE AMOUNT OF FUNCTIONING RENAL TISSUE. PMID- 16693744 TI - DISTRIBUTION OF JAUNDICE IN CIRCULATORY FAILURE. PMID- 16693745 TI - STUDIES ON THE VELOCITY OF BLOOD FLOW: II. The Velocity of Blood Flow in Normal Resting Individuals, and a Critique of the Method Used. PMID- 16693746 TI - INORGANIC SULPHATES IN HUMAN BLOOD. PMID- 16693747 TI - PREFERENTIAL UTILIZATION OF CARBOHYDRATE IN DIABETES. PMID- 16693749 TI - INVESTIGATION RELATIVE TO B. WELCHII INFECTION OF THE INTESTINAL TRACT AS THE ETIOLOGICAL FACTOR IN PERNICIOUS ANEMIA. PMID- 16693748 TI - PULMONARY GAS DIFFUSION IN POLYCYTHEMIA VERA. PMID- 16693750 TI - STUDIES UPON OBESITY: THE SOURCE OF HEAT DURING PERIODS OF REDUCTION. PMID- 16693751 TI - STUDIES ON THE VELOCITY OF BLOOD FLOW: III. The Velocity of Blood Flow and its Relation to other Aspects of the Circulation in Patients with Rheumatic and Syphilitic Heart Disease. PMID- 16693752 TI - STUDIES ON THE VELOCITY OF BLOOD FLOW: IV. The Velocity of Blood Flow and Its Relation to Other Aspects of the Circulation in Patients with Arteriosclerosis and in Patients with Arterial Hypertension. PMID- 16693753 TI - STUDIES ON THE VELOCITY OF BLOOD FLOW: V. The Physiological and the Pathological Significance of the Velocity of Blood Flow. PMID- 16693754 TI - ALTERATIONS IN LIVER FUNCTION AS AN INDEX OF TOXEMIA IN PNEUMOCOCCUS LOBAR PNEUMONIA. PMID- 16693755 TI - THE EFFECT OF EPINEPHRIN ON THE PARTITION OF FOOD STUFFS IN OBESE AND NORMAL INDIVIDUALS. PMID- 16693756 TI - THE EXCRETION OF ALBUMIN AND GLOBULIN IN NEPHRITIS. PMID- 16693757 TI - THE INFLUENCE OF EDEMA ON THE CAPACITY OF BLOOD PLASMA TO MODIFY FLUID IMBIBITION BY MUSCLE. PMID- 16693758 TI - OXYGEN CONSUMPTION, OXYGEN DEBT AND LACTIC ACID IN CIRCULATORY FAILURE. PMID- 16693759 TI - THE CONCENTRATION OF UREA IN THE BLOOD OF NORMAL INDIVIDUALS. PMID- 16693760 TI - STUDIES ON HUMAN CAPILLARIES: IV. Observations on the Nature of the Capillary Pulse in Aortic Insufficiency. PMID- 16693761 TI - STUDIES ON HUMAN CAPILLARIES: V. Observations in Cases of Heart Disease with Regular Rhythm. PMID- 16693763 TI - REPEATED BLOOD SUGAR CURVES IN NON-DIABETIC SUBJECTS. PMID- 16693762 TI - THE MECHANISM OF THE ACTION OF IODIDES ON THE NITROGEN METABOLISM. PMID- 16693764 TI - ABNORMAL SPECIFIC DYNAMIC ACTION OF PROTEIN, GLUCOSE, AND FAT ASSOCIATED WITH UNDERNUTRITION. PMID- 16693765 TI - STUDIES ON THE VELOCITY OF BLOOD FLOW: VI. The Method of Collecting the Active Deposit of Radium and Its Preparation for Intravenous Injection. PMID- 16693766 TI - STUDIES ON THE VELOCITY OF BLOOD FLOW: VII. The Pulmonary Circulation Time in Normal Resting Individuals. PMID- 16693767 TI - A METHOD FOR THE DEMONSTRATION OF CALIBRE CHANGES IN THE BRONCHI IN NORMAL RESPIRATION. PMID- 16693768 TI - GENERALIZED MYCOSIS DUE TO A HITHERTO UNDESCRIBED FUNGUS (GLENOSPORA GAMMELI). PMID- 16693769 TI - THE RATE OF GASTRIC SECRETION IN MAN. PMID- 16693770 TI - AN ADAPTATION OF THE THERMAL CONDUCTIVITY METHOD TO THE ANALYSIS OF RESPIRATORY GASES. PMID- 16693771 TI - STUDIES IN EXPERIMENTAL ANEMIA: I. The Effects on Rabbits of the Injection of the Hemolytic Toxin of the Welch Bacillus. PMID- 16693772 TI - STUDIES IN EXPERIMENTAL ANEMIA: II. The Effect on Rabbits of the Injection of Stool Extracts of Patients with Pernicious Anemia and Normal Individuals. PMID- 16693773 TI - STUDIES IN EXPERIMENTAL ANEMIA: III. An Immunologic Study of the Relation between Pernicious Anemia and Anemia Due to Welch Bacillus Toxin. PMID- 16693774 TI - QUANTITATIVE PETTENKOFER VALUES IN BLOOD WITH SPECIAL REFERENCE TO HEPATIC DISEASE: A Preliminary Report. PMID- 16693775 TI - STUDIES ON THE VELOCITY OF BLOOD FLOW: VIII. The Velocity of Blood Flow and Its Relation to Other Aspects of the Circulation in Patients with Pulmonary Emphysema. PMID- 16693776 TI - Francis Weld Peabody. PMID- 16693777 TI - THE BLOOD-URINE UREA CONCENTRATION RATIO IN HYPERTENSION. PMID- 16693779 TI - THE EFFECT OF DIGITALIS UPON THE OUTPUT OF THE HEART IN NORMAL MAN. PMID- 16693778 TI - THE OXYGEN SATURATION OF HEMOGLOBIN IN THE ARTERIAL BLOOD OF EXERCISING PATIENTS. PMID- 16693780 TI - A STUDY OF EXPERIMENTAL ANEMIA IN DOGS: THE ACTION OF BEEF LIVER AND IRON SALTS ON HEMOGLOBIN REGENERATION. PMID- 16693781 TI - ELLIPTICAL AND SICKLE-SHAPED ERYTHROCYTES IN THE CIRCULATING BLOOD OF WHITE PERSONS. PMID- 16693782 TI - THE EFFECT OF INSULIN, PITUITRIN AND ADRENALIN ON THE BLOOD-SUGAR LEVEL. PMID- 16693783 TI - CHRONIC NEPHRITIS WITH AND WITHOUT EDEMA: A STUDY OF CHOLESTEROL IN THESE CONDITIONS. PMID- 16693784 TI - RELATIONSHIP OF ACUTE INFECTIONS TO GLOMERULAR NEPHRITIS. PMID- 16693785 TI - CIRCULATORY ADJUSTMENT IN ANEMIA. PMID- 16693786 TI - THE SURFACE TENSION OF THE BLOOD SERUM IN HYPERTHYROIDISM. PMID- 16693787 TI - THE PLASMA FATS AND THE IODINE ABSORPTION CAPACITY OF THE FATTY ACIDS IN HYPERTHYROIDISM. PMID- 16693788 TI - THE CARDIODYNAMIC CHANGES IN THE AORTA AND LEFT VENTRICLE DUE TO STENOSIS OF THE AORTA. PMID- 16693789 TI - BLOOD VOLUME IN FEVER. PMID- 16693790 TI - BLOOD VOLUME IN NORMAL INFANTS AND CHILDREN. PMID- 16693791 TI - A COMPARATIVE STUDY OF THE EFFECTS OF VARIOUS TREATMENTS ON THE CALCIUM AND PHOSPHORUS METABOLISM IN TETANY: I. Chronic Juvenile Tetany. PMID- 16693792 TI - A COMPARATIVE STUDY OF THE EFFECTS OF VARIOUS TREATMENTS ON THE CALCIUM AND PHOSPHORUS METABOLISM IN TETANY: II. Chronic Adult Idiopathic Tetany. PMID- 16693793 TI - GASTRIC ACIDITY: RELATION TO VARIOUS FACTORS SUCH AS AGE AND PHYSICAL FITNESS. PMID- 16693794 TI - GASTRIC MOTILITY AND THE VOLUME OF GASTRIC SECRETION IN MAN. PMID- 16693795 TI - A COMPARATIVE STUDY OF THE ROTATORY AND REDUCING PROPERTIES OF PLASMA ULTRAFILTRATES FROM DIABETIC AND NEPHRITIC PATIENTS. PMID- 16693797 TI - STUDIES ON THE EFFECT OF CARDIAC IRREGULARITY ON THE CIRCULATION: II. The Estimation of Cardiac Output in Dogs Subject to Artificial Auricular Fibrillation. PMID- 16693796 TI - STUDIES ON THE EFFECT OF CARDIAC IRREGULARITY ON THE CIRCULATION: I. The Relation of Pulse Deficit to Rate of Blood Flow in Dogs Subject to Artificial Auricular Fibrillation and to Regular Tachycardia. PMID- 16693799 TI - CLINICAL STUDIES ON THE VELOCITY OF BLOOD FLOW: X. The Relation Between the Velocity of Blood Flow, the Venous Pressure and the Vital Capacity of the Lungs in Fifty Patients with Cardiovascular Disease Compared With Similar Measurements in Fifty Normal Persons. PMID- 16693798 TI - CLINICAL STUDIES ON THE VELOCITY OF BLOOD FLOW: IX. The Pulmonary Circulation Time, the Velocity of Venous Blood Flow to the Heart, and Related Aspects of the Circulation in Patients with Cardiovascular Disease. PMID- 16693801 TI - THE CIRCULATING BLOOD VOLUME IN DIABETIC ACIDOSIS. PMID- 16693800 TI - THE DETERMINATION OF THE CIRCULATING BLOOD VOLUME WITH CARBON MONOXIDE. PMID- 16693802 TI - THE EFFECT OF HIGH PROTEIN DIETS ON THE REMAINING KIDNEY OF RATS. PMID- 16693803 TI - THE EFFECT OF ANTI-RHEUMATIC DRUGS ON THE ARTHRITIS AND IMMUNE BODY PRODUCTION IN SERUM DISEASE. PMID- 16693804 TI - LOW BASAL METABOLISM FOLLOWING THYROTOXICOSIS: I. Temporary Type without Myxedema, with Special Reference to the Role of Iodine Therapy. PMID- 16693805 TI - LOW BASAL METABOLISM FOLLOWING THYROTOXICOSIS: II. Permanent Type without Myxedema. PMID- 16693806 TI - A NOTE ON THE INFLUENCE OF THE CIRCULATION ON THE UTILIZATION OF CARBOHYDRATES. PMID- 16693807 TI - GASTRIC FUNCTION IN CASES OF GASTRIC AND DUODENAL ULCER. PMID- 16693808 TI - THE INFLUENCE OF A DIET, HIGH IN BUTTER FAT, ON GROWTH, BLOOD FORMATION AND BLOOD DESTRUCTION. PMID- 16693809 TI - A NEW PROCEDURE FOR DETERMINING BLOOD SEDIMENTATION RATES. PMID- 16693810 TI - STUDIES IN BLOOD COMPOSITION OF ANIMALS UNDER PATHOLOGICAL CONDITIONS: I. Broncho pneumonia in cows. PMID- 16693811 TI - STUDIES OF THE CHEMICAL MECHANISM OF HYDROCHLORIC ACID SECRETION: I. Electrolyte Variations in Human Gastric Juice. PMID- 16693812 TI - STUDIES OF THE CHEMICAL MECHANISM OF HYDROCHLORIC ACID SECRETION: II. Observations on the Blood Passing through the Stomach of Dogs. PMID- 16693813 TI - THE EFFECT OF POSTURE UPON THE COMPOSITION AND VOLUME OF THE BLOOD IN MAN. PMID- 16693814 TI - THE EFFECT OF POSTURE UPON THE VELOCITY OF BLOOD FLOW IN MAN. PMID- 16693815 TI - THE CHLORIDE, BASE AND NITROGEN CONTENT OF GASTRIC JUICE AFTER HISTAMINE STIMULATION. PMID- 16693816 TI - CLINICAL STUDIES ON THE VELOCITY OF BLOOD FLOW: XI. The Pulmonary Circulation Time, the Minute Volume Blood Flow through the Lungs, and the Quantity of Blood in the Lungs. PMID- 16693817 TI - OBSERVATIONS ON THE EXCRETION OF CALCIUM IN TWO CASES OF NEPHROSIS TREATED WITH PARATHYROID EXTRACT. PMID- 16693818 TI - CHEMICAL CHANGES OCCURRING IN THE BODY AS A RESULT OF CERTAIN DISEASES IN INFANTS AND CHILDREN: II. Acute Hemorrhagic Nephritis. Sub-Acute Nephritis; Severe Chronic Nephritis. PMID- 16693819 TI - THE REDUCTION OF HYPERCALCEMIA IN CASES OF POLYCYTHEMIA VERA BY PHENYLHYDRAZINE. PMID- 16693820 TI - STUDIES OF SERUM ELECTROLYTES: III. In Infections, Nephritis, and Other Pathological Conditions. PMID- 16693821 TI - THE RELATION BETWEEN CARDIAC SIZE AND CARDIAC OUTPUT PER MINUTE FOLLOWING THE ADMINISTRATION OF DIGITALIS IN NORMAL DOGS. PMID- 16693822 TI - THE RELATION BETWEEN CARDIAC SIZE AND CARDIAC OUTPUT PER MINUTE FOLLOWING THE ADMINISTRATION OF DIGITALIS IN DOGS IN WHICH THE HEART IS ENLARGED. PMID- 16693824 TI - THE EFFECT OF MORPHINE ON THE RESPIRATION IN PNEUMONIA. PMID- 16693823 TI - THE RELATION OF ANOXEMIA TO THE TYPE OF BREATHING IN PNEUMONIA: A Study of Respiration by Means of a Body Plethysmograph. PMID- 16693825 TI - ANOXEMIA IN PNEUMONIA AND ITS RELIEF BY OXYGEN INHALATION. PMID- 16693826 TI - STUDIES ON PATHOGENESIS AND RECOVERY IN ERYSIPELAS. PMID- 16693827 TI - THE EFFECT OF ATROPINE UPON THE OUTPUT OF THE HEARTS OF NORMAL MEN. PMID- 16693828 TI - A NOTE ON THE CARDIAC OUTPUT OF A SINGLE INDIVIDUAL OBSERVED OVER A PERIOD OF FIVE YEARS. PMID- 16693829 TI - THE PROTEIN CONTENT OF THE CEREBROSPINAL FLUID IN MYXEDEMA. PMID- 16693830 TI - CHEMICAL CHANGES OCCURRING IN THE BODY AS THE RESULT OF CERTAIN DISEASES: III. The Composition of the Plasma in Severe Diabetic Acidosis and the Changes Taking Place during Recovery. PMID- 16693831 TI - THE ENERGY EXCHANGE IN OBESITY. PMID- 16693832 TI - ON THE GASEOUS EXCHANGE FOLLOWING THE ADMINISTRATION OF DIHYDROXYACETONE. PMID- 16693833 TI - ON THE SIGNIFICANCE OF RESPIRATORY QUOTIENTS AFTER ADMINISTRATION OF CERTAIN CARBOHYDRATES. PMID- 16693834 TI - TOLYSIN IN SUBACUTE RHEUMATIC CARDITIS. PMID- 16693835 TI - THE FUNCTION OF THE KIDNEYS IN PATIENTS SUFFERING FROM CHRONIC CARDIAC DISEASE WITHOUT SIGNS OF HEART FAILURE. PMID- 16693836 TI - TEMPORARY AND PERMANENT MYXEDEMA FOLLOWING TREATED AND UNTREATED THYROTOXICOSIS. PMID- 16693837 TI - GUANIDINE RETENTION AND CALCIUM RESERVE AS ANTAGONISTIC FACTORS IN CARBON TETRACHLORIDE AND CHLOROFORM POISONING. PMID- 16693838 TI - STUDIES ON DUODENAL REGURGITATION. I. PMID- 16693839 TI - STUDIES OF UREA EXCRETION. II: Relationship Between Urine Volume and the Rate of Urea Excretion by Normal Adults. PMID- 16693840 TI - STUDIES OF UREA EXCRETION. III: The Influence of Body Size on Urea Output. PMID- 16693842 TI - STUDIES OF UREA EXCRETION. V: The Diurnal Variation of Urea Excretion in Normal Individuals and Patients with Bright's Disease. PMID- 16693841 TI - STUDIES OF UREA EXCRETION. IV: Relationship Between Urine Volume and Rate of Urea Excretion by Patients with Bright's Disease. PMID- 16693843 TI - TOTAL ACID-BASE EQUILIBRIUM OF PLASMA IN HEALTH AND DISEASE: X. The Acidosis of Nephritis. PMID- 16693844 TI - TOTAL ACID-BASE EQUILIBRIUM OF PLASMA IN HEALTH AND DISEASE: XI. Hypochloremia and Total Salt Deficiency in Nephritis. PMID- 16693845 TI - TOTAL ACID-BASE EQUILIBRIUM OF PLASMA IN HEALTH AND DISEASE: XII. A Study of Renal Edema. PMID- 16693846 TI - THE INFLUENCE OF IODINE ON THE EXCRETION OF CREATINE IN EXOPHTHALMIC GOITER. PMID- 16693847 TI - THE VITAMIN B CONTENT OF CANCER. PMID- 16693848 TI - STUDIES ON DIGITALIS IN AMBULATORY CARDIAC PATIENTS: II. The Elimination of Digitalis in Man. PMID- 16693849 TI - THE OUTPUT OF THE HEART IN PATIENTS WITH ABNORMAL BLOOD PRESSURES. PMID- 16693850 TI - THE EFFECT OF THE DIGITALIS BODIES ON THE VELOCITY OF BLOOD FLOW THROUGH THE LUNGS AND ON OTHER ASPECTS OF THE CIRCULATION. A STUDY OF NORMAL SUBJECTS AND PATIENTS WITH CARDIOVASCULAR DISEASE. PMID- 16693851 TI - STUDIES OF CALCIUM AND PHOSPHORUS METABOLISM: IV. The Effect of the Parathyroid Hormone. PMID- 16693852 TI - OBSERVATIONS ON PAROXYSMAL HEMOGLOBINURIA. PMID- 16693853 TI - A QUANTITATIVE METHOD FOR THE ESTIMATION OF PEPSIN. PMID- 16693854 TI - QUANTITATIVE MEASUREMENTS OF PEPSIN IN GASTRIC JUICE BEFORE AND AFTER HISTAMINE STIMULATION. PMID- 16693855 TI - STUDIES OF CALCIUM AND PHOSPHORUS METABOLISM : II. The Calcium Excretion of Normal Individuals on a Low Calcium Diet, also Data on a Case of Pregnancy. PMID- 16693856 TI - STUDIES OF CALCIUM AND PHOSPHORUS METABOLISM: III. The Effects of the Thyroid Hormone and Thyroid Disease. PMID- 16693857 TI - STUDIES ON THE PHYSIOLOGY OF THE PARATHYROID GLANDS: I. Calcium and Phosphorus Studies on a Case of Idiopathic Hypoparathyroidism. PMID- 16693860 TI - THE INVOLUTION OF CUTANEOUS XANTHOMATA CAUSED BY DIETS LOW IN CALORIES. PMID- 16693858 TI - THE DETERMINATION OF THE CIRCULATING BLOOD VOLUME IN INFANTS BY THE CARBON MONOXIDE METHOD. PMID- 16693859 TI - AN APPARATUS FOR THE PROLONGED ADMINISTRATION OF ARTIFICIAL RESPIRATION: I. A Design for Adults and Children. PMID- 16693861 TI - THE SURFACE TENSION OF BLOOD SERUM, AND THE DETERMINATION OF THE SURFACE TENSION OF BIOLOGICAL FLUIDS. PMID- 16693862 TI - BLOOD VOLUME PRECEDING AND FOLLOWING SPLENECTOMY IN HEMOLYTIC ICTERUS AND SPLENIC ANEMIA. PMID- 16693863 TI - STUDIES OF SERUM ELECTROLYTES: IV. The Chloride and Nitrogen Balances, and Weight Changes in Pneumonia. PMID- 16693864 TI - STUDIES OF SERUM ELECTROLYTES: V. Urinary Electrolyte Excretion in Pneumonia. PMID- 16693865 TI - THE EFFECT OF EXERCISE ON THE SIZE OF NORMAL HEARTS AND OF ENLARGED HEARTS OF DOGS. PMID- 16693866 TI - METABOLISM OF CHLORIDE AND TOTAL FIXED BASE IN PNEUMONIA AND THE RELATION TO SALT AND WATER RETENTION. PMID- 16693867 TI - AN INVESTIGATION OF VARIOUS FACTORS WHICH AFFECT THE SEDIMENTATION RATE OF THE RED BLOOD CELLS. PMID- 16693868 TI - OBSERVATIONS ON THE EFFECT OF VARIOUS FACTORS ON THE DURATION OF THE ELECTRICAL SYSTOLE OF THE HEART AS INDICATED BY THE LENGTH OF THE Q-T INTERVAL OF THE ELECTROCARDIOGRAM. PMID- 16693869 TI - THE CALORIGENETIC ACTION OF THYROXIN AT DIFFERENT LEVELS OF BASAL METABOLISM IN MYXEDEMA. PMID- 16693870 TI - THE ACTION OF SODIUM CHLORIDE, AMMONIUM CHLORIDE, AND SODIUM BICARBONATE ON THE TOTAL ACID-BASE BALANCE OF A CASE OF CHRONIC NEPHRITIS WITH EDEMA. PMID- 16693871 TI - THE INFLUENCE OF THE SYMPATHETIC NERVOUS SYSTEM ON THE CAPILLARIES DURING PASSIVE CONGESTION. PMID- 16693872 TI - THE EXCRETION OF ZINC IN HEALTH AND DISEASE. PMID- 16693873 TI - SKIN REACTIONS TO FILTRATES OF HAEMOLYTIC STREPTOCOCCI IN ACUTE AND SUBACUTE NEPHRITIS. PMID- 16693874 TI - THE COLOR OF THE SKIN AS ANALYZED BY SPECTROPHO-TOMETRIC METHODS: I. Apparatus and Procedures. PMID- 16693875 TI - THE COLOR OF THE SKIN AS ANALYZED BY SPECTRO-PHOTOMETRIC METHODS: II. The Role of Pigmentation. PMID- 16693876 TI - THE COLOR OF THE SKIN AS ANALYZED BY SPECTROPHOTOMETRIC METHODS: III. The Role of Superficial Blood. PMID- 16693877 TI - STUDIES ON RED BLOOD CELL DIAMETER: III. The Relative Diameter of Immature (Reticulocytes) and Adult Red Blood Cells in Health and Anemia, Especially in Pernicious Anemia. PMID- 16693878 TI - STUDIES ON RED BLOOD CELL DIAMETER: IV. The Decrease in the Mean Diameter of the Reticulocytes and Adult Red Blood Cells in Pernicious Anemia Following Liver Therapy. PMID- 16693879 TI - THE INFLUENCE OF RESPIRATION ON VENOUS PRESSURE. PMID- 16693880 TI - GLUCOSE EXCRETION IN BRIGHT'S DISEASE. PMID- 16693881 TI - STUDIES OF THE CIRCULATION IN THREE CASES OF MORBUS CAERULEUS. PMID- 16693882 TI - VENOUS PRESSURE IN GENERAL ANESTHESIA. PMID- 16693883 TI - VARIATIONS IN THE CARDIAC OUTPUT OF MAN: V. The Cardiac Output of Man During the Malaise and Pyrexia Following the Injection of Typhoid Vaccine. PMID- 16693884 TI - AN APPARATUS FOR THE PROLONGED ADMINISTRATION OF ARTIFICIAL RESPIRATION: II. A Design for Small Children and Infants with an Appliance for the Administration of Oxygen and Carbon Dioxide. PMID- 16693885 TI - THE MEASUREMENT OF CAPILLARY PRESSURE UNDER NATURAL CONDITIONS AND AFTER ARTERIOLAR DILATATION; IN NORMAL SUBJECTS AND IN PATIENTS WITH ARTERIAL HYPERTENSION AND WITH ARTERIOSCLEROSIS. PMID- 16693886 TI - THE ENDOGENOUS URIC ACID METABOLISM IN PERNICIOUS ANEMIA. PMID- 16693887 TI - EXPERIMENTS ON THE PATENCY OF THE BLOOD VESSELS OF NEPHRITIC KIDNEYS OBTAINED AT AUTOPSY. PMID- 16693889 TI - THE DETERMINATION OF THE TOTAL HEAT ELIMINATED BY THE HUMAN BEING. PMID- 16693888 TI - SECONDARY ALTERATIONS IN TOTAL SERUM CALCIUM AFTER THE ADMINISTRATION OF GLUCOSE AND INSULIN. PMID- 16693890 TI - MEASUREMENT OF TOTAL WATER EXCHANGE. PMID- 16693891 TI - THE NATURE OF OBESITY. PMID- 16693893 TI - A CASE OF OSTEITIS FIBROSA CYSTICA (OSTEOMALACIA?) WITH EVIDENCE OF HYPERACTIVITY OF THE PARA-THYROID BODIES. METABOLIC STUDY II. PMID- 16693892 TI - A CASE OF OSTEITIS FIBROSA CYSTICA (OSTEOMALACIA?) WITH EVIDENCE OF HYPERACTIVITY OF THE PARA-THYROID BODIES. METABOLIC STUDY I. PMID- 16693894 TI - A CASE OF OSTEITIS FIBROSA CYSTICA (OSTEOMALACIA?) WITH EVIDENCE OF HYPERACTIVITY OF THE PARATHYROID BODIES. METABOLIC STUDY III. PMID- 16693896 TI - STUDIES IN CONGESTIVE HEART FAILURE: II. The Respiratory Exchange During and After Exercise. PMID- 16693895 TI - STUDIES IN CONGESTIVE HEART FAILURE: I. The Effect of Edema on Oxygen Utilization. PMID- 16693897 TI - STUDIES IN CONGESTIVE HEART FAILURE: III. The Buffering Power of the Blood and Tissues. PMID- 16693898 TI - STUDIES IN CONGESTIVE HEART FAILURE: IV. The Potassium Content of Skeletal and Cardiac Muscle. PMID- 16693899 TI - THE RELATIONSHIPS BETWEEN PLASMA SPECIFIC GRAVITY, PLASMA PROTEIN CONTENT AND EDEMA IN NEPHRITIS. PMID- 16693901 TI - STUDIES ON ICTERUS NEONATORUM: The Production of icterus in Animals Following Prolonged Anoxaemia. PMID- 16693900 TI - STUDIES OF UREA EXCRETION: VI. Comparison of the Blood Urea Clearance with Certain Other Measures of Renal Function. PMID- 16693902 TI - THE USE OF THEOCALCIN IN THE TREATMENT OF HEART FAILURE OF THE CONGESTIVE TYPE. PMID- 16693903 TI - LIVER FUNCTION IN CONGESTIVE HEART FAILURE. PMID- 16693904 TI - CIRCULATORY MEASUREMENTS IN PATIENTS WITH RHEUMATIC HEART DISEASE BEFORE AND AFTER THE ADMINISTRATION OF DIGITALIS. PMID- 16693905 TI - THE CIRCULATORY CHANGES AFTER FULL THERAPEUTIC DOSES OF DIGITALIS, WITH A CRITICAL DISCUSSION OF VIEWS ON CARDIAC OUTPUT. PMID- 16693906 TI - FURTHER OBSERVATIONS ON THE CIRCULATORY ACTIONS OF DIGITALIS AND STROPHANTHUS WITH SPECIAL REFERENCE TO THE LIVER, AND COMPARISONS WITH HISTAMINE AND EPINEPHRINE. PMID- 16693907 TI - THE PATHOLOGICAL EFFECTS UPON RATS OF EXCESS IRRADIATED ERGOSTEROL. PMID- 16693908 TI - ON THE RELATION OF CHANGES IN BLOOD VELOCITY AND VOLUME FLOW OF BLOOD TO CHANGE OF POSTURE. PMID- 16693909 TI - A METHOD FOR CORRECTING THE ERYTHROCYTE SEDIMENTATION RATE FOR VARIATIONS IN THE CELL VOLUME PERCENTAGE OF BLOOD. PMID- 16693910 TI - THE REACTION OF HUMAN DUODENAL CONTENTS TO ACID AND ALKALINE MEAT MIXTURES. PMID- 16693911 TI - STUDIES OF THE KIDNEY IN ACUTE INFECTION: I. Observations with the Urine Sediment Count (Addis) in Acute Rheumatic Infection. PMID- 16693912 TI - THE PLASMA PROTEINS IN RELATION TO BLOOD HYDRATION: IV. Malnutrition and the Serum Proteins. PMID- 16693913 TI - THE PLASMA PROTEINS IN RELATION TO BLOOD HYDRATION: V. Serum Proteins and Malnutritional or Cachectic Edema. PMID- 16693915 TI - STUDIES ON PNEUMONIA IN CHILDREN: I. Mortality, Blood Cultures, and Humoral Antibodies in Pneumococcus Pneumonia. PMID- 16693914 TI - A DYNAMIC METHOD FOR MEASUREMENT OF MUSCLE TONUS IN MAN. PMID- 16693916 TI - THE INTRADERMAL SALT TEST IN FEVER. PMID- 16693917 TI - THE DIAGNOSTIC VALUE. OF DETERMINATIONS OF PEPSIN IN GASTRIC JUICE. PMID- 16693918 TI - OBSERVATIONS ON THE SURFACE CAPILLARIES IN MAN FOLLOWING CERVICOTHORACIC SYMPATHETIC GANGLIONECTOMY. PMID- 16693919 TI - ICTERUS NEONATORUM: III. The Oxygen Capacity and Saturation of the Mother and Foetus. PMID- 16693920 TI - THE FUNCTIONAL PATHOLOGY OF HYPER-PARATHYROIDISM. PMID- 16693921 TI - THE ASSOCIATION OF RENAL AND GASTRIC DISORDERS WITH CONSTANCY OF THE URINARY REACTION. PMID- 16693922 TI - THE PNEUMOCOCCIDAL POWER OF WHOLE BLOOD: I. Description of Method. Results in Individuals with No History of Lobar Pneumonia. PMID- 16693923 TI - THE PNEUMOCOCCIDAL POWER OF WHOLE BLOOD: II. Estimations in Lobar Pneumonia. PMID- 16693924 TI - STUDIES ON THE VELOCITY OF BLOOD FLOW: XIII. The Circulatory Response to Thyrotoxicosis. PMID- 16693925 TI - STUDIES ON THE VELOCITY OF BLOOD FLOW: XIV. The Circulation in Myxedema with a Comparison of the Velocity of Blood Flow in Myxedema and Thyrotoxicosis. PMID- 16693927 TI - THE ACTION OF HISTAMINE ON THE PANCREAS. PMID- 16693926 TI - STUDIES IN CONGESTIVE HEART FAILURE: V. The Potassium Content of Skeletal Muscle Obtained by Biopsy. PMID- 16693928 TI - A COMPARISON OF THE UREA NITROGEN CONTENT OF CUTANEOUS AND VENOUS BLOOD BY MICRO GASOMETRIC ANALYSIS. PMID- 16693929 TI - FURTHER OBSERVATIONS ON EXPERIMENTAL AORTIC INSUFFICIENCY: II. Cinematographic Studies of Changes in Ventricular Size and in Left Ventricular Discharge. PMID- 16693930 TI - THE MEASUREMENT OF SYMPATHETIC VASOCONSTRICTOR ACTIVITY IN THE LOWER EXTREMITIES. PMID- 16693931 TI - SYMPATHETIC INHIBITION OF THE LARGE INTESTINE IN HIRSCHSPRUNG'S DISEASE. PMID- 16693932 TI - THE SERUM CALCIUM IN POLYCYTHEMIA VERA. PMID- 16693933 TI - SERUM ELECTROLYTE STUDIES IN NORMAL AND PATHOLOGICAL CONDITIONS: PNEUMONIA, RENAL EDEMA, CARDIAC EDEMA, UREMIC AND DIABETIC ACIDOSIS. PMID- 16693934 TI - THE COMPOSITION OF THE BILE FOLLOWING THE RELIEF OF BILIARY OBSTRUCTION. PMID- 16693935 TI - STUDIES ON THE NITROGEN AND SULPHUR METABOLISM IN "BRIGHT'S DISEASE": I. The Retention of Nitrogen and Sulphur in "Nephrosis". PMID- 16693937 TI - BLOOD VOLUME AND PLASMA ELECTROLYTE CHANGES IN THE DEHYDRATION OF INFANTS. PMID- 16693936 TI - THE EFFECT OF ATROPINE UPON GASTRIC SECRETION AFTER HISTAMINE STIMULATION. PMID- 16693938 TI - THE PROBLEM OF SODIUM SALICYLATE EXCRETION IN THE BILE. PMID- 16693939 TI - THE TOLERANCE OF NORMAL SUBJECTS TO LEVULOS E: Factors Influencing the Variations in Rise in Blood Sugar. PMID- 16693940 TI - NUCLEOTIDE THERAPY IN AGRANULOCYTOSIS. PMID- 16693941 TI - STUDIES IN CONGESTIVE HEART FAILURE: VI. The Effect of Overwork and Other Factors on the Potassium Content of the Cardiac Muscle. PMID- 16693942 TI - STUDIES IN CONGESTIVE HEART FAILURE: VII. The Effect of Overwork on the Potassium Content of Skeletal Muscle. PMID- 16693944 TI - THE VARIATIONS OF THE SPECIFIC GRAVITY OF THE PLASMA OF THE BLOOD AND THE MEANS AVAILABLE FOR ALTERING IT. PMID- 16693943 TI - THE NUMBER OF FORMED ELEMENTS IN THE URINARY SEDIMENT OF PATIENTS SUFFERING FROM HEART DISEASE, WITH PARTICULAR REFERENCE TO THE STATE OF HEART FAILURE. PMID- 16693945 TI - THE OPTICAL ACTIVITY OF CEREBROSPINAL FLUID IN SUPPURATIVE MENINGITIS, AND ITS LACTIC ACID, SUGAR, AND CHLORIDE CONTENT. PMID- 16693946 TI - THE PROPORTION OF CERTAIN IMPORTANT INORGANIC CONSTITUENTS IN THE DYING HEART MUSCLE. PMID- 16693947 TI - CARBON DIOXIDE AND OXYGEN TENSIONS OF THE MIXED VENOUS BLOOD OF MAN AT REST. PMID- 16693948 TI - LIVER EXTRACT, LIVER ASH AND IRON IN THE TREATMENT OF ANEMIA. PMID- 16693949 TI - THE RELATIONSHIP OF BLOOD URIC ACID CONTENT TO THE STATE OF RENAL FUNCTION IN NEPHRITIS. PMID- 16693950 TI - PHYSIOLOGICAL STUDIES OF FAINTNESS AND SYNCOPE. PMID- 16693951 TI - THE EXCRETION OF INTRAVENOUSLY INJECTED BILIRUBIN AS A TEST OF LIVER FUNCTION. PMID- 16693952 TI - CHANGES IN BLOOD VOLUME IN PATIENTS WITH EDEMA OF RENAL ORIGIN. PMID- 16693953 TI - EFFECT OF ACIDOSIS AND ALKALOSIS UPON CAPACITY FOR WORK. PMID- 16693954 TI - STUDIES OF SERUM ELECTROLYTES: VII. The Total Base and Protein Components of the Serum during Lobar Pneumonia with a Note on the Gastric Secretion. PMID- 16693955 TI - THE ELIMINATION OF ETHYL IODIDE AFTER INHALATION AND ITS RELATION TO THERAPEUTIC ADMINISTRATION. PMID- 16693956 TI - NORMAL STANDARDS OF GASTRIC FUNCTION. PMID- 16693957 TI - STUDIES ON THE PHYSIOLOGY OF THE PARATHYROID GLANDS: II. The Relation of the Serum Calcium to the Serum Phosphorus at Different Levels of Parathyroid Activity. PMID- 16693958 TI - STUDIES ON THE VELOCITY OF BLOOD FLOW: XV. The Velocity of Blood Flow and Other Aspects of the Circulation in Patients with "Primary" and Secondary Anemia and in Two Patients with Polycythemia Vera. PMID- 16693959 TI - STUDIES IN CONGESTIVE HEART FAILURE: VIII. The Effect of the Administration of Dibasic Potassium Phosphate on the Potassium Content of Certain Tissues. PMID- 16693960 TI - BLOOD ELECTROLYTE STUDIES DURING HISTAMINE SHOCK IN DOGS. PMID- 16693961 TI - THE PATHOLOGIC PHYSIOLOGY OF PELLAGRA: V. The Circulating Blood Volume. PMID- 16693962 TI - SKIN REACTIONS OF PATIENTS AND NORMAL INDIVIDUALS TO PROTEIN EXTRACTS OF STREPTOCOCCI. PMID- 16693963 TI - STUDIES IN CONGESTIVE HEART FAILURE: IX. The Effect of Digitalis on the Potassium Content of the Cardiac Muscle of Dogs. PMID- 16693964 TI - THE HEART RATE OF BOYS DURING AND AFTER EXHAUSTING EXERCISE. PMID- 16693965 TI - HIGH NITROGEN DIETS AND RENAL INJURY: The Dependence of the Injury upon the Nature of the Nitrogenous Substance. PMID- 16693966 TI - STUDIES OF CALCIUM AND PHOSPHORUS METABOLISM: VIII. The Influence of the Thyroid Gland and the Parathyroid Hormone upon the Total Acid-Base Metabolism. PMID- 16693967 TI - RESPONSES TO THE INJECTION OF EPINEPHRINE IN HEPATIC DISEASE. PMID- 16693968 TI - THE SPREAD OF RHEUMATIC FEVER THROUGH FAMILIES. PMID- 16693970 TI - THE PATHOLOGIC PHYSIOLOGY OF PELLAGRA: I. Tabulated Clinical and Physiologic Data. PMID- 16693969 TI - AGE SUSCEPTIBILITY TO FAMILIAL INFECTION IN RHEUMATIC FEVER. PMID- 16693971 TI - THE PATHOLOGIC PHYSIOLOGY OF PELLAGRA: II. The Serum Albumin and Globulin. PMID- 16693972 TI - THE PATHOLOGIC PHYSIOLOGY OF PELLAGRA: III. The Serum Calcium and Phosphorus, with Especial Reference to Nervous Symptoms. PMID- 16693973 TI - STUDIES ON GALLBLADDER FUNCTION: II. The Absorption of Sodium Tetraiodophenolphthalein from the Gallbladder. PMID- 16693974 TI - THE PATHOLOGIC PHYSIOLOGY OF PELLAGRA: IV. Serum Electrolytes and Acid-Base Equilibrium. PMID- 16693975 TI - STUDIES OF CALCIUM AND PHOSPHORUS METABOLISM: XII. The Effect of the Ingestion of Acid-producing Substances. PMID- 16693976 TI - STUDIES OF CALCIUM AND PHOSPHORUS METABOLISM: XIII. The Effect of Ingestion of Phosphates on the Excretion of Calcium. PMID- 16693977 TI - STUDIES OF CALCIUM AND PHOSPHORUS METABOLISM: XVIII. On Temporary Fluctuations in the Level of Calcium and Inorganic Phosphorus in Blood Serum of Normal Individuals. PMID- 16693978 TI - GASTRIC SECRETION AFTER HISTAMINE: SODIUM AND POTASSIUM CONTENT AND PEPSIN ESTIMATION. PMID- 16693979 TI - STUDIES ON THE NITROGEN AND SULPHUR METABOLISM IN "BRIGHT'S DISEASE": II. Observations on the Nitrogen and Sulphur Excretion in Patients without Renal Edema. PMID- 16693980 TI - ICTERUS NEONATORUM: IV. The Role of the Placenta in Visible Icterus Neonatorum. PMID- 16693981 TI - STREPTOCOCCAL AGGLUTININS IN CHRONIC INFECTIOUS ARTHRITIS. PMID- 16693982 TI - STREPTOCOCCAL AGGLUTININS IN RHEUMATIC FEVER. PMID- 16693983 TI - STUDIES OF THE KIDNEY IN ACUTE INFECTION: II. Observations with the Urea Clearance Test in Acute Rheumatic Infection. PMID- 16693984 TI - STUDIES OF THE KIDNEY IN ACUTE INFECTION: III. Observations with the Urine Sediment Count (Addis) and the Urea Clearance Test in Lobar Pneumonia. PMID- 16693985 TI - BODY BUILD IN INFANTS: I. The Technique of Measuring the External Dimensions of the Body in Infants. AB - In these three papers the body build of infants during health and disease is described quantitatively. This is done by comparison of ratios of various external dimensions to the total body length. In the first paper the technique for measuring the external dimensions of the body in infants is described and various sources of error discussed. In the second paper general empirical formulae for the relationship between various external dimensions and total body length in healthy infants are developed. By comparing two groups of infants from different social environments, it is shown that environment may influence body build. In the third paper the results obtained in the second paper for healthy infants are used for comparison with sick infants. Various differences in the body build of infants with acute intestinal intoxication, tetany and eczema are described. PMID- 16693986 TI - BODY BUILD IN INFANTS: II. The Proportions of the External Dimensions of the Healthy Infant during the First Year of Life. PMID- 16693987 TI - BODY BUILD IN INFANTS: III. Body Build in Disease. PMID- 16693988 TI - THE MECHANICAL INFLUENCE OF THE PERICARDIUM UPON CARDIAC FUNCTION. PMID- 16693989 TI - STUDIES ON COLLATERAL CIRCULATION. I. THERMIC CHANGES AFTER ARTERIAL LIGATION AND GANGLIONECTOMY. PMID- 16693990 TI - STUDIES ON COLLATERAL CIRCULATION. II. THERMIC CHANGES AFTER ARTERIAL LIGATION, SECTION OF SPINAL CORD OR POSTERIOR ROOTS AND GANGLIONECTOMY. PMID- 16693992 TI - EXPERIMENTAL REFERRED PAIN FROM THE GASTRO-INTESTINAL TRACT. PART II. STOMACH, DUODENUM AND COLON. PMID- 16693991 TI - EXPERIMENTAL REFERRED PAIN FROM THE GASTRO-INTESTINAL TRACT. PART I. THE ESOPHAGUS. PMID- 16693993 TI - THE BLOOD VOLUME IN HYPERTHYROIDISM. PMID- 16693994 TI - THE ACID-BASE EQUILIBRIUM IN PATHOLOGICAL CONDITIONS: II. ALKALOSIS OBSERVED IN HYPERTENSIVE STATES. PMID- 16693996 TI - THE EFFECT OF INSULIN HYPOGLYCEMIA ON THE CIRCULATION. PMID- 16693995 TI - A STUDY OF GLYCOLYSIS. PMID- 16693997 TI - STUDIES ON THE RELATIONSHIP BETWEEN OXYGEN CONSUMPTION AND NITROGEN METABOLISM. I. IN PERNICIOUS ANEMIA. PMID- 16693998 TI - BREATHING MEASUREMENTS ON NORMAL NEWBORN INFANTS. PMID- 16693999 TI - COLLATERAL RESPIRATION. TRANSFER OF AIR COLLATERALLY BETWEEN PULMONARY LOBULES. PMID- 16694000 TI - THE HEALING OF RICKETS COINCIDENT WITH LOW SERUM INORGANIC PHOSPHORUS. PMID- 16694001 TI - A QUASI-CONTINUOUS RECORDER FOR OXYGEN AND CARBON DIOXIDE FOR CLINICAL ATMOSPHERE CONTROL. PMID- 16694002 TI - AN ELECTROCARDIOGRAPHIC STUDY OF THE HEART IN LOBAR PNEUMONIA. PMID- 16694003 TI - THE RELATIONSHIP BETWEEN THE ENVIRONMENT AND THE BASAL INSENSIBLE LOSS OF WEIGHT. PMID- 16694004 TI - A METHOD FOR THE DETERMINATION OF HEAT PRODUCTION OVER LONG PERIODS OF TIME. PMID- 16694005 TI - AN IMPROVED METHOD FOR THE DETERMINATION OF WATER BALANCE. PMID- 16694006 TI - THE DOUBTFUL NATURE OF "LUXUSKONSUMPTION". PMID- 16694007 TI - THE DETERMINATION OF POTASSIUM IN CARDIAC MUSCLE AND THE PRESUMABLE INFLUENCE OF THE BETA RADIATIONS ON THE RHYTHM. PMID- 16694008 TI - GASTRIC SECRETION AFTER HISTAMINE: ITS EVALUATION AS A QUANTITATIVE TEST IN DIGESTIVE DISORDERS. PMID- 16694009 TI - MINERAL EXCHANGES OF MAN: III. Mineral Metabolism During Treatment of a Case of Polycythemia Vera. PMID- 16694010 TI - INSULIN ABSORPTION BY THE CONJUNCTIVAL MEMBRANES IN RABBITS. PMID- 16694011 TI - ELECTROCARDIOGRAPHIC STUDIES DURING ATTACKS OF ANGINA PECTORIS AND OF OTHER PAROXYSMAL PAIN. PMID- 16694012 TI - STUDIES IN CONGESTIVE HEART FAILURE: XIII. The Relation of Dyspnea of Exertion to the Oxygen Saturation and Acid-Base Condition of the Blood. PMID- 16694013 TI - STUDIES IN CONGESTIVE HEART FAILURE: XIV. Orthopnea: Its Relation to Ventilation, Vital Capacity, Oxygen Saturation and Acid-Base Condition of Arterial and Jugular Blood. PMID- 16694014 TI - A COMPARISON OF THE RACEMIZATION CURVES FOR URINARY, EDEMA FLUID, AND BLOOD PLASMA PROTEINS. PMID- 16694015 TI - NUTRITIONAL EDEMA. OBSERVATIONS ON THE RELATION OF THE SERUM PROTEINS TO THE OCCURRENCE OF EDEMA AND TO THE EFFECT OF CERTAIN INORGANIC SALTS. PMID- 16694016 TI - STUDIES ON THE ELECTRICAL SYSTOLE ("Q-T" INTERVAL) OF THE HEART: II. Its Duration in Cardiac Failure. PMID- 16694017 TI - THE EFFECTS OF SUSTAINED PITUITARY ANTIDIURESIS AND FORCED WATER DRINKING IN EPILEPTIC CHILDREN. A DIAGNOSTIC AND ETIOLOGIC STUDY. PMID- 16694018 TI - THE PLASMA PROTEINS IN RELATION TO BLOOD HYDRATION. VI. SERUM PROTEINS IN NEPHRITIC EDEMA. PMID- 16694019 TI - STUDIES ON THE MODE OF ACTION OF IRRADIATED ERGOSTEROL: I. Its effect on the Calcium, Phosphorus and Nitrogen Metabolism of Normal Individuals. PMID- 16694020 TI - THE PLASMA PROTEINS IN RELATION TO BLOOD HYDRATION: VIII. Serum Proteins in Heart Disease. PMID- 16694021 TI - THE PLASMA PROTEINS IN RELATION TO BLOOD HYDRATION: IX. Serum Proteins in the Terminal Stages of Renal Disease. PMID- 16694022 TI - THE VARIATIONS IN SERUM CALCIUM, PROTEIN, AND INORGANIC PHOSPHORUS IN EARLY AND LATE PREGNANCY, DURING PARTURITION AND THE PUERPERIUM, AND IN NON-PREGNANT WOMEN. PMID- 16694023 TI - THE EFFECT OF LIVER AND COMMERCIAL LIVER EXTRACT ON THE BODY WEIGHT, RED BLOOD CELLS, AND RETICULOCYTES OF NORMAL RATS. PMID- 16694024 TI - STUDIES IN CONGESTIVE HEART FAILURE: XV. Reflex Versus Chemical Factors in the Production of Rapid Breathing. PMID- 16694025 TI - GASTRIC ACIDITY IN NORMAL INDIVIDUALS. PMID- 16694026 TI - THE GASTRIC SECRETION IN EXOPHTHALMIC GOITRE AND MYXOEDEMA. PMID- 16694027 TI - EFFECTS OF ENVIRONMENTAL TEMPERATURE, ANESTHESIA AND LUMBAR SYMPATHETIC GANGLIONECTOMY ON THE TEMPERATURES OF THE EXTREMITIES OF ANIMALS. PMID- 16694029 TI - OBSERVATIONS ON THE FORMATION OF WHEALS: V. The Effects of Variation of the CO(2) Combining Power of the Blood on Histamine Wheals. PMID- 16694028 TI - OBSERVATIONS ON THE FORMATION OF WHEALS: IV. The Influence of Calcium Concentrations on Histamine Wheals. PMID- 16694030 TI - STUDIES ON THE MODE OF ACTION OF IRRADIATED ERGOSTEROL: II. Its Effect on the Calcium and Phosphorus Metabolism of Individuals with Calcium Deficiency Diseases. PMID- 16694031 TI - STUDIES OF CALCIUM AND PHOSPHORUS METABOLISM: VI. In Hypoparathyroidism and Chronic Steatorrhea with Tetany with Special Consideration of the Therapeutic Effect of Thyroid. PMID- 16694032 TI - STUDIES OF CALCIUM AND PHOSPHORUS METABOLISM: XV. In Various Metabolic and Bone Diseases. PMID- 16694033 TI - STUDIES ON THE MODE OF ACTION OF IRRADIATED ERGOSTEROL: III. The Effect of Irradiated Ergosterol Administration on the Formation of Bone Trabeculae. PMID- 16694034 TI - STUDIES ON THE MODE OF ACTION OF IRRADIATED ERGOSTEROL: IV. in Hypoparathyroidism. PMID- 16694035 TI - THE MOVEMENT OF FLUID THROUGH THE HUMAN CAPILLARY WALL IN RELATION TO VENOUS PRESSURE AND TO THE COLLOID OSMOTIC PRESSURE OF THE BLOOD. PMID- 16694036 TI - THE PLASMA PROTEINS IN RELATION TO BLOOD HYDRATION: VII. A Note on the Proteins in Acute Nephritis. PMID- 16694037 TI - INTRAVENOUS INJECTIONS: A Study of the Composition of the Blood During Continuous Trauma to the Intestines When No Fluid is Injected and When Fluid is Injected Continuously. PMID- 16694038 TI - INTRAVENOUS INJECTIONS: A Study of the Effects on the Composition of the Blood of the Injection of Various Fluids into Dogs with Normal and with Low Blood Pressures. PMID- 16694039 TI - A STUDY OF THE EFFECTS OF HEMORRHAGE, TRAUMA, HISTAMINE AND SPINAL ANESTHESIA ON THE COMPOSITION OF THE BLOOD WHEN NO FLUIDS ARE INJECTED AND WHEN FLUIDS ARE INTRODUCED INTRAVENOUSLY. PMID- 16694040 TI - THE EFFECTS ON THE COMPOSITION OF THE BLOOD OF THE SUBCUTANEOUS INJECTION OF NORMAL SALT SOLUTION INTO NORMAL DOGS AND INTO DOGS SUBJECTED TO INTESTINAL TRAUMA, GRADED HEMORRHAGES AND HISTAMINE INJECTION. PMID- 16694041 TI - STUDIES IN THE METABOLISM OF SODIUM r-LACTATE. I. RESPONSE OF NORMAL HUMAN SUBJECTS TO THE INTRAVENOUS INJECTION OF SODIUM r-LACTATE. PMID- 16694042 TI - STUDIES IN THE METABOLISM OF SODIUM r-LACTATE. II. RESPONSE OF HUMAN SUBJECTS WITH ACIDOSIS TO THE INTRAVENOUS INJECTION OF SODIUM r-LACTATE. PMID- 16694043 TI - STUDIES IN THE METABOLISM OF SODIUM r-LACTATE. III. RESPONSE OF HUMAN SUBJECTS WITH LIVER DAMAGE, DISTURBED WATER AND MINERAL BALANCE, AND RENAL INSUFFICIENCY TO THE INTRAVENOUS INJECTION OF SODIUM r-LACTATE. PMID- 16694044 TI - A CELLULOID CAPSULE FOR MEASURING VENOUS PRESSURES. PMID- 16694045 TI - CUTANEOUS RESPIRATION IN MAN: IV. The Rate of Carbon Dioxide Elimination and Oxygen Absorption in Normal Subjects. PMID- 16694046 TI - CUTANEOUS RESPIRATION IN MAN: V. The Rate of Carbon Dioxide Elimination and Oxygen Absorption in Subjects with Diseases of the Skin. PMID- 16694047 TI - CUTANEOUS RESPIRATION IN MAN: VI. The Effect of Drugs on the Rate of Carbon Dioxide Elimination and Oxygen Absorption. PMID- 16694048 TI - CUTANEOUS RESPIRATION IN MAN: VII. The Effect of Venous Congestion on the Rate of Carbon Dioxide Elimination and Oxygen Absorption. PMID- 16694049 TI - STUDIES OF CALCIUM AND PHOSPHORUS METABOLISM: XIV. The Relation of Acid-Base Balance to Phosphate Balance Following Ingestion of Phosphates. PMID- 16694051 TI - THE SPECIFIC DYNAMIC RESPONSE TO PROTEIN OF INDIVIDUALS SUFFERING FROM DISEASE OF THE HYPOPHYSIS. PMID- 16694050 TI - STUDIES IN PARATHYROID PHYSIOLOGY: III. The Effect of Phosphate Ingestion in Clinical Hyperparathyroidism. PMID- 16694052 TI - STIMULATION OF GASTRIC PEPSIN BY HISTAMINE. PMID- 16694053 TI - CHEMICAL STUDIES OF THE BLOOD IN HIGH INTESTINAL OBSTRUCTION : I. The Distribution of Phosphorus and Intracellular Changes. PMID- 16694054 TI - CHEMICAL STUDIES OF THE BLOOD IN HIGH INTESTINAL OBSTRUCTION: II. The Relation between "Toxemia" and Chemical Changes. PMID- 16694055 TI - THE SPINAL FLUID IN HYPERTENSION. PMID- 16694056 TI - THE INFLUENCE OF EXPERIMENTAL THYROID INTOXICATION ON THE POTASSIUM, SODIUM, AND WATER CONTENT OF THE MYOCARDIUM. PMID- 16694057 TI - THE EXCRETION OF XYLOSE AS AN INDEX OF DAMAGED RENAL FUNCTION. PMID- 16694058 TI - HYPOGLYCEMIC REACTIONS FOLLOWING GLUCOSE INGESTION. PMID- 16694059 TI - THE COPPER AND NON-HEMOGLOBINOUS IRON CONTENTS OF THE BLOOD SERUM IN DISEASE. PMID- 16694060 TI - CHANGES IN THE CIRCULATION PRODUCED BY GRADUAL OCCLUSION OF THE PULMONARY ARTERY. PMID- 16694061 TI - STUDIES OF BLOOD GLYCOLYSIS: I. Sugar and Phosphorus Relationships during Glycolysis in Normal Blood. PMID- 16694062 TI - STUDIES OF BLOOD GLYCOLYSIS: II. Sugar and Phosphorus Relationships during Glycolysis in the Blood of Infants and Children with Various Diseases. PMID- 16694063 TI - INCREASED ELASTIC TENSION OF THE LUNG IN EXPERIMENTAL PNEUMONIA. PMID- 16694064 TI - THE EXCRETION OF INORGANIC SULPHATES. PMID- 16694065 TI - ON THE MECHANISM OF NEPHROTIC EDEMA. PMID- 16694066 TI - THE EFFECT OF VENTRICULIN ON THE BLOOD SUGAR LEVEL IN PATIENTS WITH PERNICIOUS ANEMIA. PMID- 16694067 TI - THE EFFECT OF INJECTION OF NONSPECIFIC PROTEIN ON THE PAIN OF ULCER AND ON GASTRIC SECRETION: A CLINICAL AND EXPERIMENTAL STUDY. PMID- 16694068 TI - THE METABOLISM OF LEUCOCYTES FROM NORMAL AND LEUKEMIC BLOOD. PMID- 16694069 TI - LIPID CONTENT OF TUMORS. PMID- 16694070 TI - PLASMAPHERESIS EDEMA. I. THE RELATION OF REDUCTION OF SERUM PROTEINS TO EDEMA AND THE PATHOLOGICAL ANATOMY ACCOMPANYING PLASMAPHERESIS. PMID- 16694071 TI - PLASMAPHERESIS EDEMA. II. THE EFFECT OF REDUCTION OF SERUM PROTEIN ON THE ELECTROLYTE PATTERN AND CALCIUM CONCENTRATION. PMID- 16694072 TI - THE PASSAGE OF FLUID AND PROTEIN THROUGH THE HUMAN CAPILLARY WALL DURING VENOUS CONGESTION. PMID- 16694073 TI - A STUDY OF THE PROTEIN-LIPID COMBINATIONS IN BLOOD AND BODY FLUIDS: I. Normal Human and Dog Plasma and Horse Serum. PMID- 16694074 TI - THE BLOOD URIC ACID IN DISEASE. PMID- 16694075 TI - A STUDY OF THE GASTRIC SECRETION IN HYPERTHYROIDISM BEFORE AND AFTER OPERATION. PMID- 16694076 TI - THYROID HYPERPLASIA PRODUCED IN CHICKENS BY ULTRAVIOLET LIGHT DEFICIENCY. PMID- 16694077 TI - LACTIC ACID IN THE BLOOD OF RESTING MAN. PMID- 16694078 TI - A STUDY OF THE CIRCULATION IN OBESITY. PMID- 16694079 TI - A COMPARATIVE STUDY OF THE EXCRETION OF WATER AND SOLIDS BY NORMAL AND ABNORMAL KIDNEYS. PMID- 16694080 TI - STUDIES ON THE PHYSIOLOGY OF THE PARATHYROID GLANDS: V. Action of Parathyroid Extract on the Renal Threshold for Phosphorus. PMID- 16694081 TI - VASODILATATION IN THE LOWER EXTREMITIES IN RESPONSE TO IMMERSING THE FOREARMS IN WARM WATER. PMID- 16694082 TI - THE EFFECT OF NASO-PHARYNGEAL OPERATIONS ON RENAL FUNCTION. PMID- 16694083 TI - STUDIES OF UREA EXCRETION. VII THE EFFECTS OF POSTURE AND EXERCISE ON UREA EXCRETION. PMID- 16694084 TI - STUDIES ON THE EFFECT OF THE ACTION OF DIGITALIS ON THE OUTPUT OF BLOOD FROM THE HEART: I. The Effect on the Output of the Dog's Heart in Heart-Lung Preparations. PMID- 16694085 TI - STUDIES ON THE EFFECT OF THE ACTION OF DIGITALIS ON THE OUTPUT OF BLOOD FROM THE HEART: II. The Effect on the Output of the Hearts of Dogs Subject to Artificial Auricular Fibrillation. PMID- 16694086 TI - STUDIES ON THE EFFECT OF THE ACTION OF DIGITALIS ON THE OUTPUT OF BLOOD FROM THE HEART. III. : Part 1. The Effect on the Output in Normal Human Hearts, Part 2. The Effect on the Output of Hearts in Heart Failure with Congestion, in Human Beings. PMID- 16694087 TI - THE LACTIC ACID OF THE BLOOD IN HEPATIC DISEASE. PMID- 16694088 TI - DIETARY PROTEIN IN HEMORRHAGIC BRIGHT'S DISEASE: I. Effects upon the Course of the Disease with Special Reference to Hematuria and Renal Function. PMID- 16694089 TI - STUDIES OF DIABETES MELLITUS: Evidence That the Disability is Concerned Solely with the Metabolism of Glucose. The Mode of Action of Insulin. PMID- 16694090 TI - STUDIES ON THE COURSE OF VASOMOTOR FIBERS AS MEASURED BY THERMIC CHANGES IN THE FEET AFTER ARTERIAL LIGATION AND SECTION OF THE SPINAL CORD AT VARIOUS LEVELS. PMID- 16694092 TI - THE LUNG VOLUME AND ITS SUBDIVISIONS: I. METHODS OF MEASUREMENT. PMID- 16694091 TI - STUDIES ON THE ACTION OF DIURETICS I. THE EFFECT OF EUPHYLLIN AND SALYRGAN UPON GLOMERULAR FILTRATION AND TUBULAR REABSORPTION. PMID- 16694093 TI - THE UREA CLEARANCE TEST IN TOXEMIAS OF PREGNANCY. PMID- 16694094 TI - GASTRO-INTESTINAL STUDIES. I. GASTRIC JUICE IN PERNICIOUS ANEMIA. PMID- 16694095 TI - THE BLOOD FLOW IN THE BRAIN AND THE LEG OF MAN, AND THE CHANGES INDUCED BY ALTERATION OF BLOOD GASES. PMID- 16694096 TI - THE BLOOD PICTURE IN EXOPHTHALMIC GOITRE AND ITS CHANGES RESULTING FROM IODINE AND OPERATION. A STUDY BY MEANS OF THE SUPRAVITAL TECHNIQUE. PMID- 16694098 TI - THE EFFECT OF THE HEART'S POSITION ON THE ELECTROCARDIOGRAPHIC APPEARANCE OF VENTRICULAR EXTRASYSTOLES. PMID- 16694097 TI - EXPERIMENTAL OBSERVATIONS ON THE EFFECT OF VARIOUS DIURETICS WHEN INJECTED DIRECTLY INTO ONE RENAL ARTERY OF THE DOG. PMID- 16694099 TI - STUDIES ON THE ELECTRICAL SYSTOLE ("Q-T" INTERVAL) OF THE HEART: IV. The Effect of Digitalis on Its Duration in Cardiac Failure. PMID- 16694100 TI - STUDIES OF SERUM ELECTROLYTES: VIII. The Concentration of Electrolytes and Non Electrolytes in the Serum Following Insulin Administration in Diabetic Patients. PMID- 16694101 TI - STUDIES OF CALCIUM AND PHOSPHORUS METABOLISM. XX. THE HIGH CALCIUM EXCRETION IN EXOPHTHALMIC GOITER IS NOT DUE TO VITAMINE D DEFICIENCY. PMID- 16694102 TI - OBSERVATIONS ON INTRAPLEURAL PRESSURE AND ITS INFLUENCE ON THE RELATIVE CIRCULATION RATE IN EMPHYSEMA. PMID- 16694103 TI - QUANTITATIVE ASPECTS OF IRON DEFICIENCY IN HYPOCHROMIC ANEMIA: (The Parenteral Administration of Iron). PMID- 16694104 TI - THE ROLE OF CALCIUM, PHOSPHORUS AND VITAMIN D IN PREGNANCY. PMID- 16694105 TI - STUDIES ON CHEYNE-STOKES RESPIRATION. PMID- 16694106 TI - STUDIES ON GASTRIC SECRETION. PMID- 16694107 TI - THE EFFECTS OF TEMPERATURE AND OF TISSUE PRESSURE ON THE MOVEMENT OF FLUID THROUGH THE HUMAN CAPILLARY WALL. PMID- 16694108 TI - STUDIES OF THE BASAL WORK AND OUTPUT OF THE HEART IN CLINICAL CONDITIONS. PMID- 16694109 TI - A NOTE ON CUTANEOUS VENOUS BLOOD SUGAR DIFFERENCE IN NORMAL MALES AND FEMALES AND IN THYROID DISEASE. PMID- 16694110 TI - INEQUALITY OF BLOOD PRESSURE IN THE BRACHIAL ARTERIES, WITH ESPECIAL REFERENCE TO DISEASE OF THE ARCH OF THE AORTA. PMID- 16694111 TI - GASTRIC SECRETION IN FEVER AND INFECTIOUS DISEASES. PMID- 16694112 TI - COLLATERAL RESPIRATION: Spontaneous Reinflation of an Atelectatic Pulmonary Lobule by Collateral Respiration. PMID- 16694113 TI - ATTEMPTS TO PRODUCE EXPERIMENTAL GASTRITIS. PMID- 16694114 TI - THE RELATION BETWEEN PLASMA PROTEIN CONTENT, PLASMA SPECIFIC GRAVITY AND EDEMA IN DOGS MAINTAINED ON A PROTEIN INADEQUATE DIET AND IN DOGS RENDERED EDEMATOUS BY PLASMAPHERESIS. PMID- 16694115 TI - THE EFFECT OF SERUM TRANSFUSION ON THE PLASMA PROTEIN DEPLETION ASSOCIATED WITH NUTRITIONAL EDEMA IN DOGS. PMID- 16694116 TI - A STUDY OF THE ETIOLOGICAL RELATIONSHIP BETWEEN PELLAGRA AND PERNICIOUS ANEMIA. PMID- 16694117 TI - THE RATES OF UTILIZATION OF THYROXINE AND OF DESICCATED THYROID IN MAN: THE RELATION BETWEEN THE IODINE IN DESICCATED THYROID AND IN THYROXINE. PMID- 16694118 TI - THE TREATMENT OF AGRANULOCYTOSIS WITH ADENINE SULPHATE. PMID- 16694119 TI - STUDIES IN SO-CALLED WATER INTOXICATION. PMID- 16694121 TI - THE UNAVOIDABLE ERROR IN THE DIFFERENTIAL COUNT OF THE LEUKOCYTES OF THE BLOOD. PMID- 16694120 TI - STUDIES ON GALLBLADDER FUNCTION. IX. THE ANION-CATION CONTENT OF BILE FROM THE NORMAL AND INFECTED GALLBLADDER. PMID- 16694122 TI - THE ADDIS SEDIMENT COUNT IN NORMAL CHILDREN. PMID- 16694123 TI - THE ADDIS SEDIMENT COUNT IN SCARLET FEVER. PMID- 16694124 TI - THE LIPIDS OF THE BLOOD PLASMA IN EPILEPSY: I. A Statistical Study of Single Determinations in 100 Epileptic and 32 "Normal" Subjects. PMID- 16694125 TI - THE LIPIDS OF THE BLOOD PLASMA IN EPILEPSY: II. Variations of Lipids in Relation to Occurrence of Seizures. PMID- 16694126 TI - STREPTOCOCCAL AGGLUTININS IN PATIENTS WITH RHEUMATOID (ATROPHIC) ARTHRITIS AND ACUTE RHEUMATIC FEVER. PMID- 16694127 TI - SKIN REACTIONS TO NUCLEOPROTEIN OF STREPTOCOCCUS SCARLATINAE IN PATIENTS WITH RHEUMATOID ARTHRITIS AND RHEUMATIC FEVER. PMID- 16694128 TI - THE SEDIMENTATION RATE OF THE RED BLOOD CELLS IN VARIOUS TYPES OF ARTHRITIS. PMID- 16694129 TI - ON DIABETIC ACIDOSIS: A Detailed Study of Electrolyte Balances Following the Withdrawal and Reestablishment of Insulin Therapy. PMID- 16694130 TI - THE CALORIGENIC ACTION OF THYROXIN POLYPEPTIDE. PMID- 16694131 TI - HYPERVENTILATION IN ARTERIOLAR HYPERTENSION. PMID- 16694132 TI - ANEMIA OF INFANCY FROM MATERNAL IRON DEFICIENCY IN PREGNANCY. PMID- 16694133 TI - SERUM PROTEINS IN DIABETIC ACIDOSIS. PMID- 16694134 TI - THE NATURE OF DIABETIC ACIDOSIS. PMID- 16694135 TI - EXPERIMENTAL PNEUMOCOCCUS LOBAR PNEUMONIA IN THE DOG : I. Method of Production and Course of the Disease. PMID- 16694136 TI - EXPERIMENTAL PNEUMOCOCCUS LOBAR PNEUMONIA IN THE DOG: II. Pathology. PMID- 16694137 TI - EXPERIMENTAL PNEUMOCOCCUS LOBAR PNEUMONIA IN THE DOG: III. Pathogenesis. PMID- 16694138 TI - PLASMA PROTEIN AND PLASMA COLLOID OSMOTIC PRESSURE IN PATHOLOGICAL CONDITIONS WITH SPECIAL REFERENCE TO THE OCCURRENCE OF EDEMA. PMID- 16694139 TI - FURTHER STUDIES ON THE AGGLUTINATION REACTION IN CHRONIC ARTHRITIS. PMID- 16694140 TI - GASTRO-INTESTINAL STUDIES. II. PANCREATIC ENZYMES IN PERNICIOUS ANEMIA. PMID- 16694141 TI - HEAT CRAMPS. A CLINICAL AND CHEMICAL STUDY. PMID- 16694142 TI - STUDIES OF GASTRIC PEPSIN. I. METHODS OF MEASUREMENT AND FACTORS WHICH INFLUENCE IT. PMID- 16694143 TI - STUDIES OF GASTRIC PEPSIN. II. SECRETION OF PEPSIN IN CASES OF DUODENAL ULCER AND PSEUDO-ULCER. PMID- 16694144 TI - STUDIES OF UREA EXCRETION. VIII. THE EFFECTS ON THE UREA CLEARANCE OF CHANGES IN PROTEIN AND SALT CONTENTS OF THE DIET. PMID- 16694145 TI - THERMAL CHANGES IN PERIPHERAL VASCULAR DISEASE DURING SYMPATHETIC GANGLIONECTOMY UNDER GENERAL ANESTHESIA. PMID- 16694146 TI - GLYCOLYSIS IN THE BLOOD OF PATIENTS WITH PERNICIOUS ANEMIA. PMID- 16694147 TI - PLASMA PROTEIN CHANGES AND SUSPENSION STABILITY OF THE BLOOD IN LOBAR PNEUMONIA. PMID- 16694148 TI - THE BLOOD IN CASES OF UNEXPLAINED GASTRIC ANACIDITY. PMID- 16694149 TI - THE INFLUENCE OF MINERAL METABOLISM UPON NEPHROTIC EDEMA. PMID- 16694150 TI - THE MEASURED EFFECT OF LAPAROTOMY ON THE RESPIRATION. PMID- 16694151 TI - EFFECT OF LAPAROTOMY ON LUNG VOLUME. DEMONSTRATION OF A NEW TYPE OF PULMONARY COLLAPSE. PMID- 16694152 TI - A STUDY OF HUMAN LIVER BILE AFTER RELEASE OF COMMON DUCT OBSTRUCTION. PMID- 16694153 TI - THE AGGLUTINATING PROPERTIES OF EXUDATES FROM PATIENTS WITH RHEUMATIC FEVER. PMID- 16694154 TI - THE ROLE OF THYROXIN IODINE AND TOTAL ORGANIC IODINE IN THE CALORIGENIC ACTION OF WHOLE THYROID GLAND. PMID- 16694155 TI - A NOTE ON THE CALCULATION OF WATER EXCHANGE. PMID- 16694156 TI - THE NATURE OF "PREFORMED WATER". PMID- 16694157 TI - LEUKOCYTOSIS FOLLOWING THE INTRAMUSCULAR INJECTION OF LIVER EXTRACT. PMID- 16694158 TI - THE EFFECT OF THYROXIN UPON THE METABOLISM OF ISOLATED NORMAL AND MALIGNANT TISSUE. PMID- 16694159 TI - THE ACTION OF CERTAIN DIURETICS ON THE FUNCTION OF THE KIDNEY AS MEASURED BY THE UREA CLEARANCE TEST. PMID- 16694160 TI - STUDIES ON THE ACTION OF DIURETICS. II. THE EFFECT OF SALYRGAN UPON THE WATER CONTENT OF THE PLASMA AS MEASURED BY THE REFRACTIVE INDEX. PMID- 16694161 TI - STUDIES IN CONGESTIVE HEART FAILURE. XXIIII. A CRITICAL STUDY OF METHODS FOR DETERMINING THE CARDIAC OUTPUT IN PATIENTS WITH CARDIAC DISEASE. PMID- 16694162 TI - PHYSIOLOGICAL DISTURBANCES DURING EXPERIMENTAL DIPHTHERITIC INTOXICATION. I. BLOOD SUGAR, LACTIC ACID AND NON-PROTEIN AND AMINO-ACID NITROGEN. PMID- 16694163 TI - PHYSIOLOGICAL DISTURBANCES DURING EXPERIMENTAL DIPHTHERITIC INTOXICATION. II. HEPATIC GLYCO-GENESIS AND GLYCOGEN CONCENTRATION OF CARDIAC AND SKELETAL MUSCLE. PMID- 16694164 TI - PHYSIOLOGICAL DISTURBANCES DURING EXPERIMENTAL DIPHTHERITIC INTOXICATION. III. RESPIRATORY QUOTIENTS AND METABOLIC RATE. PMID- 16694165 TI - STUDIES OF TOTAL PULMONARY CAPACITY AND ITS SUB-DIVISIONS. I. NORMAL, ABSOLUTE AND RELATIVE VALUES. PMID- 16694166 TI - STUDIES OF TOTAL PULMONARY CAPACITY AND ITS SUB-DIVISIONS. II. CORRELATION WITH PHYSICAL AND RADIOLOGICAL MEASUREMENTS. PMID- 16694167 TI - STUDIES OF TOTAL PULMONARY CAPACITY AND ITS SUB-DIVISIONS. III. CHANGES WITH BODY POSTURE. PMID- 16694168 TI - STUDIES OF TOTAL PULMONARY CAPACITY AND ITS SUB-DIVISIONS. IV. PRELIMINARY OBSERVATIONS ON CASES OF PULMONARY EMPHYSEMA AND OF PNEUMOCONIOSIS. PMID- 16694169 TI - THE EFFECT OF DIGITALIS ON THE VENOUS PRESSURE OF NORMAL INDIVIDUALS. PMID- 16694170 TI - A COMPARISON OF THE CREATININE AND UREA CLEARANCE TESTS OF KIDNEY FUNCTION. PMID- 16694172 TI - SERUM TREATMENT OF HEMOLYTIC STREPTOCOCCUS PNEUMONIA. PMID- 16694171 TI - EXPERIMENTS ON THE RELATION OF CREATININE AND UREA CLEARANCE TESTS OF KIDNEY FUNCTION AND THE NUMBER OF GLOMERULI IN THE HUMAN KIDNEY OBTAINED AT AUTOPSY. PMID- 16694173 TI - CYSTINE CONTENT OF FINGER NAILS IN PELLAGRA. PMID- 16694174 TI - THE PRODUCTION OF EXPERIMENTAL PLAUT-VINCENT'S ANGINA IN THE DOG. PMID- 16694175 TI - THE EFFECTS OF ALTERNATE SUCTION AND PRESSURE ON BLOOD FLOW TO THE LOWER EXTREMITIES. PMID- 16694176 TI - THE EFFECT OF SPLENIC CONTRACTION ON THE FORMED ELEMENTS OF THE BLOOD IN A CASE OF ANEMIA AND SPLENOMEGALY. PMID- 16694177 TI - THE PROTEINS OF BLOOD AND SUBCUTANEOUS LYMPH IN DOGS. PMID- 16694178 TI - PERMEABILITY OF CAPILLARIES TO PLASMA LIPOIDS. PMID- 16694179 TI - THE CLINICAL SIGNIFICANCE OF THE ERYTHROCYTIC SEDIMENTATION TEST IN RHEUMATOID ARTHRITIS. PMID- 16694180 TI - THE OXYGENATION OF CONCENTRATED VERSUS NORMAL BLOODS. PMID- 16694181 TI - ELECTROLYTES IN HUMAN TISSUE. III. A COMPARISON OF NORMAL HEARTS WITH HEARTS SHOWING CONGESTIVE HEART FAILURE. PMID- 16694182 TI - THE CISTERNAL PRESSURE IN CONGESTIVE HEART FAILURE AND ITS BEARING ON ORTHOPNEA. PMID- 16694183 TI - THE ACTION OF PHLORIZIN ON THE EXCRETION OF GLUCOSE, XYLOSE, SUCROSE, CREATININE AND UREA BY MAN. PMID- 16694184 TI - AMINO NITROGEN CHANGES OF THE BLOOD IN NEPHRITIS. PMID- 16694185 TI - A FORMULA AND NOMOGRAM FOR THE ESTIMATION OF THE OSMOTIC PRESSURE OF COLLOIDS FROM THE ALBUMIN AND TOTAL PROTEIN CONCENTRATIONS OF HUMAN BLOOD SERA. PMID- 16694186 TI - HERPETIC PHARYNGITIS AND STOMATITIS. A REPORT OF THREE CASES. PMID- 16694187 TI - STUDIES UPON THE NASAL SECRETIONS. I. THE CELLULAR CONTENT OF THE NASAL SECRETIONS IN ACUTE DISEASE OF THE RESPIRATORY TRACT. PMID- 16694188 TI - MAGNESIUM METABOLISM IN HYPERPARATHYROIDISM. PMID- 16694189 TI - IMMUNITY IN DIABETES: INFLUENCE OF DIABETES ON THE DEVELOPMENT OF ANTIBACTERIAL PROPERTIES IN THE BLOOD. PMID- 16694190 TI - A NOTE ON THE CALCULATION OF WATER EXCHANGE. PMID- 16694191 TI - THE RATE OF EXCRETION OF URINE IN SUBJECTS WITH DIFFERENT AMOUNTS OF RENAL TISSUE. PMID- 16694192 TI - SALT AND WATER IN THE TREATMENT OF DIABETIC ACIDOSIS. PMID- 16694193 TI - THE DIURNAL RHYTHM IN WATER AND MINERAL EXCHANGE. PMID- 16694194 TI - THE BLOOD LIPIDS IN THE POSTABSORPTIVE STATE AND AFTER THE INGESTION OF FAT IN NORMAL HUMAN SUBJECTS AND IN A CASE OF DISSEMINATED CUTANEOUS XANTHOMATA. PMID- 16694195 TI - ANTIBODY RESPONSE TO INFECTIONS WITH THE NEWLY CLASSIFIED TYPES OF PNEUMOCOCCI (COOPER). PMID- 16694196 TI - THE RELATION BETWEEN PHYSICAL CONSTITUTION AND THE INCIDENCE OF DISEASE: The Disease Groups Include Peptic Ulcer, Cholecystitis and Diabetes Mellitus. PMID- 16694197 TI - OBSERVATIONS UPON THE CALCIUM AND PHOSPHORUS METABOLISM IN CERTAIN DISEASES OF BONE. PMID- 16694198 TI - STUDIES ON THE RELATIONSHIP BETWEEN OXYGEN CONSUMPTION AND NITROGEN METABOLISM. III. IN POLYCYTHEMIA VERA. PMID- 16694199 TI - ANTISTREPTOLYSIN CONTENT OF THE BLOOD SERUM IN RHEUMATIC FEVER AND RHEUMATOID ARTHRITIS. PMID- 16694200 TI - STUDIES OF TOTAL PULMONARY CAPACITY AND ITS SUBDIVISIONS. V. NORMAL VALUES IN FEMALE SUBJECTS. PMID- 16694201 TI - THE EFFECT OF DIIODOTYROSINE ON THE BASAL METABOLISM IN MYXEDEMA. PMID- 16694202 TI - THE PRODUCTION OF PAIN IN EXERCISING SKELETAL MUSCLE DURING INDUCED ANOXEMIA. PMID- 16694203 TI - FIBRINOLYTIC ACTIVITY OF HEMOLYTIC STREPTOCOCCI. THE DEVELOPMENT OF RESISTANCE TO FIBRINOLYSIS FOLLOWING ACUTE HEMOLYTIC STREPTOCOCCUS INFECTIONS. PMID- 16694204 TI - ANTIBODY RESPONSE TO INFECTIONS WITH TYPE III AND THE RELATED TYPE VIII PNEUMOCOCCUS. PMID- 16694205 TI - ANTIBODY RESPONSE TO INFECTIONS WITH TYPE II AND THE RELATED TYPE V PNEUMOCOCCUS. PMID- 16694206 TI - THE DIGESTION OF BEEF PROTEINS IN THE HUMAN STOMACH. PMID- 16694207 TI - CUTANEOUS REACTIONS IN THE DIAGNOSIS OF UNDULANT FEVER. PMID- 16694208 TI - THE DISTRIBUTION OF BLOOD PHOSPHORUS AFTER SUPPRESSION OF RENAL FUNCTION. PMID- 16694209 TI - LIPOIDS OF SERUM IN DIABETIC ACIDOSIS. PMID- 16694210 TI - KIDNEY FUNCTION AND BLOOD PRESSURE. PMID- 16694211 TI - THE MEASUREMENT OF THE INTRAPLEURAL PRESSURE IN MAN AND ITS SIGNIFICANCE. PMID- 16694212 TI - THE ELASTIC PROPERTIES OF THE EMPHYSEMATOUS LUNG AND THEIR CLINICAL SIGNIFICANCE. PMID- 16694213 TI - THE INTRAPLEURAL PRESSURE IN CONGESTIVE HEART FAILURE AND ITS CLINICAL SIGNIFICANCE. PMID- 16694214 TI - THE LIPEMIA OF PREGNANCY. PMID- 16694215 TI - OBSERVATIONS ON THE CHEMICAL AND PHYSICAL RELATION BETWEEN BLOOD SERUM AND BODY FLUIDS. 1. THE NATURE OF EDEMA FLUIDS AND EVIDENCE REGARDING THE MECHANISM OF EDEMA FORMATION. PMID- 16694216 TI - THE MEASUREMENT OF THE ELASTICITY AND VISCOSITY OF SKELETAL MUSCLE IN NORMAL AND PATHOLOGICAL CASES; A STUDY OF SOCALLED "MUSCLE TONUS". PMID- 16694217 TI - THE IMMEDIATE RESPONSE OF THE PLASMA CHOLESTEROL TO THE INJECTION OF INSULIN AND OF EPINEPHRINE IN HUMAN SUBJECTS. PMID- 16694218 TI - STUDIES OF PHOSPHORUS OF BLOOD. II. THE PARTITION OF PHOSPHORUS IN BLOOD IN RELATION TO THE CORPUSCLE VOLUME. PMID- 16694219 TI - ANTIBODY RESPONSES IN INFECTIOUS MONONUCLEOSIS. PMID- 16694220 TI - A REPORT OF FAILURE TO PRODUCE GRANULOCYTOPENIA WITH BACTERIAL TOXINS. PMID- 16694221 TI - THE EFFECT OF POSTURE (STANDING) ON THE SERUM PROTEIN CONCENTRATION AND COLLOID OSMOTIC PRESSURE OF BLOOD FROM THE FOOT IN RELATION TO THE FORMATION OF EDEMA. PMID- 16694222 TI - EPINEPHRINE HYPERGLYCEMIA: With Particular Reference to the Arteriovenous Blood Sugar Difference in Hepatic Disease. PMID- 16694223 TI - THE INCIDENCE AND BIOLOGICAL CHARACTERISTICS OF THE HEMOLYTIC BACILLUS COLI IN THE STOOLS OF HEALTHY INDIVIDUALS. PMID- 16694224 TI - THE RELATION OF THE CEREBROSPINAL AND VENOUS PRESSURES IN HEART FAILURE. PMID- 16694225 TI - THE RELATION OF RHEUMATIC FEVER TO POSTSCARLATINAL ARTHRITIS AND POSTSCARLATINAL HEART DISEASE-A FAMILIAL STUDY. PMID- 16694226 TI - PASSAGE OF NATIVE PROTEINS THROUGH THE NORMAL GASTRO-INTESTINAL WALL. PMID- 16694227 TI - STUDIES IN CONGESTIVE HEART FAILURE XXII. A METHOD FOR OBTAINING "MIXED" VENOUS BLOOD BY ARTERIAL PUNCTURE. PMID- 16694228 TI - RELATIONSHIP BETWEEN BLOOD CHOLESTEROL AND INCREASED METABOLISM FROM DINITROPHENOL AND THYROID. PMID- 16694229 TI - PHYSIOLOGICAL DISTURBANCES DURING EXPERIMENTAL DIPHTHERITIC INTOXICATION. IV. BLOOD ELECTROLYTE AND HEMOGLOBIN CONCENTRATIONS. PMID- 16694231 TI - THE RADIATION OF HEAT FROM THE HUMAN BODY: I. An Instrument for Measuring the Radiation and Surface Temperature of the Skin. PMID- 16694230 TI - STUDIES OF THE HEART AND CIRCULATION IN DISEASE; ESTIMATIONS OF BASAL CARDIAC OUTPUT, METABOLISM, HEART SIZE, AND BLOOD PRESSURE IN 235 SUBJECTS. PMID- 16694232 TI - THE RADIATION OF HEAT FROM THE HUMAN BODY: II. A Comparison of Some Methods of Measurement. PMID- 16694233 TI - THE RADIATION OF HEAT FROM THE HUMAN BODY: III. The Human Skin as a Black-Body Radiator. PMID- 16694234 TI - THE RELATION OF CIRCULATING ANTIPNEUMOCOCCAL IMMUNE SUBSTANCES TO THE COURSE OF LOBAR PNEUMONIA I. NATURAL IMMUNE SUBSTANCES. PMID- 16694235 TI - THE RELATION OF CIRCULATING ANTIPNEUMOCOCCAL IMMUNE SUBSTANCES TO THE COURSE OF LOBAR PNEUMONIA II. ACQUIRED IMMUNE SUBSTANCES. PMID- 16694236 TI - THE RELATION OF CIRCULATING ANTIPNEUMOCOCCAL IMMUNE SUBSTANCES TO THE COURSE OF LOBAR PNEUMONIA III. INJECTED IMMUNE SUBSTANCES (ANTIPNEUMOCOCCUS SERUM, TYPES I AND II). PMID- 16694237 TI - RELATION OF VARIATIONS IN MEAN CORPUSCULAR VOLUME TO NUMBER OF RETICULOCYTES IN PERNICIOUS ANEMIA: The Significance of Increased Bone Marrow Activity in Determining the Mean Size of Red Corpuscles. PMID- 16694238 TI - THE DIAGNOSTIC IMPORTANCE OF THE HETEROPHILE ANTIBODY TEST IN LEUKEMIA. PMID- 16694239 TI - THE EFFECT OF CORONARY OCCLUSION UPON THE INITIAL PHASE OF THE VENTRICULAR COMPLEX IN PRECORDIAL LEADS. PMID- 16694240 TI - THE INFLUENCE OF PROTEIN INTAKE ON THE UREA CLEARANCE IN NORMAL MAN. PMID- 16694241 TI - THE EFFECTS ON RENAL ACTIVITY OF THE ORAL ADMINISTRATION OF PHLORIZIN IN MAN. PMID- 16694242 TI - ON THE RENAL EXCRETION OF XYLOSE. PMID- 16694243 TI - THE CHARACTERISTICS OF SYNOVIAL FLUID IN GONOCOCCAL ARTHRITIS. PMID- 16694244 TI - THE URINARY EXCRETION OF IODINE. I. THE LOSS OF IODINE IN THE URINE FOLLOWING THYROIDECTOMY. PMID- 16694245 TI - THERAPEUTIC EFFECT OF TOTAL ABLATION OF NORMAL THYROID ON CONGESTIVE HEART FAILURE AND ANGINA PECTORIS. IX. POSTOPERATIVE PARATHYROID FUNCTION. CLINICAL OBSERVATIONS AND SERUM CALCIUM AND PHOSPHORUS STUDIES. PMID- 16694246 TI - OBSERVATIONS ON THE TREATMENT OF PELLAGRA. PMID- 16694247 TI - THE RADIATION OF HEAT FROM THE HUMAN BODY. IV. THE EMISSION, REFLECTION, AND TRANSMISSION OF INFRA-RED RADIATION BY THE HUMAN SKIN. PMID- 16694248 TI - STUDIES OF TOTAL PULMONARY CAPACITY AND ITS SUB-DIVISIONS. VI. OBSERVATIONS ON CASES OF OBSTRUCTIVE PULMONARY EMPHYSEMA. PMID- 16694249 TI - STUDIES OF TOTAL PULMONARY CAPACITY AND ITS SUBDIVISIONS. VII. OBSERVATIONS DURING THE ACUTE RESPIRATORY DISTRESS OF BRONCHIAL ASTHMA AND FOLLOWING THE ADMINISTRATION OF EPINEPHRINE. PMID- 16694250 TI - RADIATION OF HEAT FROM THE HUMAN BODY: A STATEMENT RELATIVE TO THE CRITIQUE OF J. D. HARDY. PMID- 16694251 TI - OBSERVATIONS ON THE EFFECT OF VAGUS AND SYMPATHETIC STIMULATION ON THE CORONARY FLOW OF THE REVIVED HUMAN HEART. PMID- 16694252 TI - THE PHOSPHATASE CONTENT OF THE BLOOD SERUM IN JAUNDICE. PMID- 16694254 TI - UNEXPLAINED FEVER IN HEART FAILURE. PMID- 16694253 TI - LOCALIZATION OF PAIN ACCOMPANYING FARADIC EXCITATION OF STOMACH AND DUODENUM IN HEALTHY INDIVIDUALS. PMID- 16694255 TI - FEVER IN HEART FAILURE RELATIONS BETWEEN THE TEMPERATURES OF THE INTERIOR AND THE SURFACE OF THE BODY. PMID- 16694256 TI - THE SIGNIFICANCE OF THE VESSELS OF THE SKIN IN ESSENTIAL HYPERTENSION. PMID- 16694257 TI - THE NEUFELD METHOD OF PNEUMOCOCCUS TYPE DETERMINATION AS CARRIED OUT IN A PUBLIC HEALTH LABORATORY: A STUDY OF 760 TYPINGS. PMID- 16694258 TI - THE EFFECT ON RENAL EFFICIENCY OF LOWERING ARTERIAL BLOOD PRESSURE IN CASES OF ESSENTIAL HYPERTENSION AND NEPHRITIS. PMID- 16694259 TI - THE ACIDOSIS OF GUANIDINE INTOXICATION. PMID- 16694260 TI - THE EFFECT OF ALKALI ON THE ABSORPTION OF A PEPTIDE OF THYROXINE FROM THE GASTRO INTESTINAL TRACT. PMID- 16694261 TI - FACTORS DETERMINING THE EFFECT OF EXERCISE ON BLOOD SUGAR IN THE DIABETIC. PMID- 16694262 TI - THE LACTIC ACID AND GLUTATHIONE CONTENT OF THE BLOOD OF SCHIZOPHRENIC PATIENTS. PMID- 16694263 TI - THE SIGNIFICANCE OF CONCENTRATION AND DILUTION TESTS IN BRIGHT'S DISEASE. PMID- 16694264 TI - STUDY OF THE BLOOD IN CHRONIC RESPIRATORY DISEASES, WITH SPECIAL REFERENCE TO THE VOLUME OF THE BLOOD. PMID- 16694265 TI - THE VALIDITY OF RAPID DETERMINATIONS OF THE OSMOTIC PRESSURE OF PROTEIN SOLUTIONS. PMID- 16694266 TI - CONCERNING THE NATURALLY OCCURRING PORPHYRINS: I. The Isolation of Coproporphyrin I from the Urine in a Case of Cincophen Cirrhosis. PMID- 16694267 TI - CONCERNING THE NATURALLY OCCURRING PORPHYRINS: II. The Isolation of a Hitherto Undescribed Porphyrin Occurring with an Increased Amount of Coproporphyrin I in the Feces of a Case of Familial Hemolytic Jaundice. PMID- 16694268 TI - CONCERNING THE NATURALLY OCCURRING PORPHYRINS: III. The Isolation of Coproporphyrin I from the Feces of Untreated Cases of Pernicious Anemia. PMID- 16694269 TI - THE RESISTANCE TO FIBRINOLYTIC ACTIVITY OF THE HEMOLYTIC STREPTOCOCCUS WITH SPECIAL REFERENCE TO PATIENTS WITH RHEUMATIC FEVER AND RHEUMATOID (ATROPHIC) ARTHRITIS. PMID- 16694270 TI - ANTI-TRYPTIC ACTIVITY OF SYNOVIAL FLUID IN PATIENTS WITH VARIOUS TYPES OF ARTHRITIS. PMID- 16694271 TI - THE INHIBITION OF TRYPTIC DIGESTION OF CARTILAGE BY SYNOVIAL FLUID FROM PATIENTS WITH VARIOUS TYPES OF ARTHRITIS. PMID- 16694272 TI - THE ABILITY OF NEPHRITIC PATIENTS TO DEAMINIZE AND FORM UREA FROM INGESTED GLYCINE. PMID- 16694273 TI - VENOUS PRESSURE AND POSTURE IN NORMAL YOUNG WOMEN. PMID- 16694274 TI - A SURGICAL TREATMENT OF ESSENTIAL HYPERTENSION. PMID- 16694275 TI - THE EFFECT OF RENAL DENERVATION ON THE LEVEL OF ARTERIAL BLOOD PRESSURE AND RENAL FUNCTION IN ESSENTIAL HYPERTENSION. PMID- 16694276 TI - SOME EFFECTS OF EXERCISE ON THE URINARY SEDIMENT. PMID- 16694277 TI - THE PROTEIN CONTENT OF SUBCUTANEOUS EDEMA FLUID IN HEART DISEASE. PMID- 16694278 TI - THE CALORIGENIC ACTION OF D- AND L-THYROXIN. PMID- 16694279 TI - EXPERIMENTAL STUDY OF CLINICAL VITAMIN B DEFICIENCY. PMID- 16694280 TI - A HISTOLOGICAL STUDY OF THE ARTERIOLES OF THE MUSCLE AND SKIN FROM THE ARM AND LEG IN INDIVIDUALS WITH COARCTATION OF THE AORTA. PMID- 16694281 TI - THE METABOLIC MEASUREMENT OF THE WATER EXCHANGE. PMID- 16694282 TI - THE EXCHANGE OF LIPIDS IN THE UMBILICAL CIRCULATION AT BIRTH. PMID- 16694283 TI - THE INFLUENCE OF IONIZED AIR UPON NORMAL SUBJECTS. PMID- 16694284 TI - STUDIES OF TOTAL PULMONARY CAPACITY AND ITS SUBDIVISIONS. VIII. OBSERVATIONS ON CASES OF PULMONARY FIBROSIS. PMID- 16694285 TI - STUDIES OF TOTAL PULMONARY CAPACITY AND ITS SUBDIVISIONS IX. RELATIONSHIP TO THE OXYGEN SATURATION AND CARBON DIOXIDE CONTENT OF THE ARTERIAL BLOOD. PMID- 16694286 TI - PLASMA FATTY ACIDS AFTER ADRENALIN INJECTION IN NORMAL SUBJECTS AND IN PATIENTS WITH LIVER DISEASE: PROGNOSTIC SIGNIFICANCE. PMID- 16694287 TI - THE EXPERIMENTAL PRODUCTION OF LOSS OF HEMATOPOIETIC ELEMENTS OF THE GASTRIC SECRETION AND OF THE LIVER IN SWINE WITH ACHLORHYDRIA AND ANEMIA. PMID- 16694288 TI - APPLICABILITY OF REBREATHING METHOD FOR DETERMINING MIXED VENOUS CO(2) IN CASES OF CHRONIC PULMONARY DISEASE. PMID- 16694289 TI - CARDIAC OUTPUT IN RELATION TO UNILATERAL PNEUMOTHORAX IN MAN. PMID- 16694290 TI - CYTOLOGIC STUDIES ON RHEUMATIC FEVER: II. Cells of Rheumatic Exudates. PMID- 16694291 TI - THE EXCHANGES OF WATER, ELECTROLYTES AND HEAT DURING PHENYLETHYLHYDANTOIN SICKNESS. PMID- 16694292 TI - OBSERVATIONS UPON THE CALCIUM AND PHOSPHORUS METABOLISM IN A CASE OF ACROMEGALY SHOWING MARKED OSTEOPOROSIS. PMID- 16694294 TI - HEMOLYTIC ANTIBODIES FOR SHEEP AND OX ERYTHROCYTES IN INFECTIOUS MONONUCLEOSIS. PMID- 16694293 TI - A NOTE ON THE METABOLIC CRITERIA OF HYPERPARATHYROIDISM. PMID- 16694295 TI - STUDIES IN SERUM ELECTROLYTES. IX. THE CHANGE IN TOTAL QUANTITY AND OSMOLAL CONCENTRATION OF GLUCOSE AND CHLORIDE IN THE SERUM AFTER THE INGESTION OF GLUCOSE BY DIABETIC PATIENTS. PMID- 16694296 TI - THE WATER AND BASE BALANCE OF THE BODY. PMID- 16694297 TI - THE CHANGES IN THE DISTRIBUTION OF BODY WATER ACCOMPANYING INCREASE AND DECREASE IN EXTRACELLULAR ELECTROLYTE. PMID- 16694298 TI - THE OCCURRENCE OF ANTIFIBRINOLYTIC PROPERTIES IN THE BLOOD OF PATIENTS WITH ACUTE HEMOLYTIC STREPTOCOCCUS INFECTIONS. PMID- 16694299 TI - DUROZIEZ'S SIGN IN NORMAL SUBJECTS AND IN PATIENTS WITH ARTERIAL HYPERTENSION: With Special Reference to Its Relation to Capillary Pulsation and the Forward Flow of Blood During Diastole. PMID- 16694300 TI - AN OPTIMAL DIET IN PROMOTING NITROGEN GAIN IN NEPHROSIS. PMID- 16694301 TI - THE EFFECT OF VENESECTION ON ARTERIAL, SPINAL FLUID, AND VENOUS PRESSURES WITH ESPECIAL REFERENCE TO FAILURE OF THE LEFT AND RIGHT HEART. PMID- 16694302 TI - MICROCOCCUS TETRAGENUS INFECTION: I. Review of the Literature, Report of a Non fatal Case with Septicemia, Meningitis and Arthritis, and Bacteriologic Studies. PMID- 16694303 TI - THE FATE OF PEOPLE WITH UNEXPLAINED GASTRIC ANACIDITY: Follow-up Studies. PMID- 16694304 TI - THE RELATION OF UPPER RESPIRATORY INFECTIONS TO RHEUMATIC FEVER IN CHILDREN: I. The Significance of Hemolytic Streptococci in the Pharyngeal Flora During Respiratory Infection. PMID- 16694305 TI - THE RELATION OF UPPER RESPIRATORY INFECTIONS TO RHEUMATIC FEVER IN CHILDREN: II. Antihemolysin Titres in Respiratory Infections and Their Significance in Rheumatic Fever in Children. PMID- 16694306 TI - THE RELATION OF UPPER RESPIRATORY INFECTIONS TO RHEUMATIC FEVER IN CHILDREN: III. The Seasonal Bacterial Flora of the Throat in Rheumatic and Non-Rheumatic Children. PMID- 16694307 TI - THE VARIABILITY OF NON-HEMOGLOBIN IRON. PMID- 16694308 TI - THE EFFECT OF PROTEIN ON THE CARBOHYDRATE TOLERANCE OF TWO CASES HAVING COMBINED DIABETES MELLITUS AND PERNICIOUS ANEMIA. PMID- 16694309 TI - BLOOD PRESSURE CHANGES IN NORMALS AND IN HYPERTENSIVES AFTER INTRAVENOUS EPINEPHRINE AND HISTAMINE. PMID- 16694310 TI - PNEUMONIA DUE TO PNEUMOCOCCUS TYPE XIV (COOPER) AND ITS TREATMENT WITH SPECIFIC ANTISERUM. PMID- 16694311 TI - THE MINUTE VOLUME OUTPUT AND THE WORK OF THE HEART IN HYPOTHYROIDISM. PMID- 16694312 TI - IMMUNITY IN DIABETES. II. RELATIVE IMPORTANCE OF NUTRITIONAL STATE AND OF BLOOD SUGAR LEVEL IN INFLUENCING DEVELOPMENT OF THE AGGLUTININ AFTER TYPHOID VACCINE. PMID- 16694313 TI - THE EXCRETION OF INULIN, XYLOSE AND UREA BY NORMAL AND PHLORIZINIZED MAN. PMID- 16694314 TI - THE RENAL EXCRETION OF CREATININE IN MAN. PMID- 16694315 TI - THE ELECTROLYTE BALANCE IN ACUTE GOUT. PMID- 16694316 TI - A STUDY OF THE LEUKOCYTOSIS PRODUCED IN MAN BY ARTIFICIAL FEVER. PMID- 16694317 TI - ON AN URTICARIAL RESPONSE TO LIGHT AND ITS PHOTOPHYSIOLOGY. PMID- 16694319 TI - ON THE BEHAVIOR OF HEMOPHILUS INFLUENZAE IN CERTAIN DISEASES OF CHILDREN. PMID- 16694318 TI - THE EFFECT OF RENAL DENERVATION ON PATIENTS SUFFERING FROM NEPHRITIS. PMID- 16694320 TI - EXPERIMENTAL CARDITIS. CHANGES IN THE MYOCARDIUM AND PERICARDIUM OF RABBITS SENSITIZED TO STREPTOCOCCI. PMID- 16694321 TI - THE RATE OF ABSORPTION OF IODIDE AND GLYCINE FROM THE GASTRO-INTESTINAL TRACT IN NORMAL PERSONS AND IN DISEASE CONDITIONS. PMID- 16694322 TI - THE HEART IN THYROID DISEASE. I. THE EFFECT OF THYROIDECTOMY ON THE ORTHODIAGRAM. PMID- 16694323 TI - THE HEART IN THYROID DISEASE. II. THE EFFECT OF THYROIDECTOMY ON THE ELECTROCARDIOGRAM. PMID- 16694324 TI - STREPTOCOCCAL AGGLUTININS AND ANTISTREPTOLYSINS IN RHEUMATOID (ATROPHIC) ARTHRITIS. PMID- 16694325 TI - SULFUR METABOLISM IN CYSTINURIA. PMID- 16694326 TI - OBSERVATIONS ON SODIUM CHLORIDE RESTRICTION AND UREA CLEARANCE IN RENAL INSUFFICIENCY. PMID- 16694327 TI - VITAL CAPACITY OF THE LUNGS: CHANGES OCCURRING IN HEALTH AND DISEASE. PMID- 16694328 TI - STUDIES ON THE MECHANISM OF THE INCREASED OXYGEN CONSUMPTION IN PATIENTS WITH CARDIAC DISEASE. PMID- 16694329 TI - STUDIES OF KIDNEY FUNCTION IN CHILDREN: I. Urea Clearance Values: (1) No Evidence of Kidney Disease (2) After Acute Hematuric Nephritis Following an Acute Infection (3) in the Acute Stage of Hematuric Nephritis. PMID- 16694330 TI - KIDNEY FUNCTION DURING NORMAL PREGNANCY I. THE INCREASED UREA CLEARANCE OF NORMAL PREGNANCY. PMID- 16694331 TI - SERUM LIPOIDS IN DIABETES. PMID- 16694332 TI - SPECIFIC DYNAMIC ACTION OF CARBOHYDRATE AND OF PROTEIN IN HUMAN HYPOTHYROIDISM AFTER TOTAL ABLATION OF THE NORMAL THYROID GLAND. PMID- 16694333 TI - THE INFLUENCE OF VARYING As-Vs INTERVALS ON SPLIT FIRST HEART SOUNDS: ITS BEARING ON THE CAUSE OF SPLIT SOUNDS AND THE MECHANISM OF THE FIRST SOUND. PMID- 16694334 TI - COMPARATIVE CALORIGENIC ACTION OF NORMAL AND PATHO-LOGICAL THYROID GLANDS ADMINISTERED IN EQUI-THYROXINE DOSES. PMID- 16694335 TI - CLINICAL OBSERVATIONS ON THE EVENTS PRECEDING THE APPEARANCE OF RHEUMATIC FEVER. PMID- 16694336 TI - FURTHER OBSERVATIONS ON THE LEUKOCYTIC RESPONSE INDUCED BY THE INTRAMUSCULAR INJECTION OF LIVER EXTRACT. PMID- 16694337 TI - THE RELATIONSHIP BETWEEN INSENSIBLE WATER LOSS AND HEAT PRODUCTION IN PATIENTS WITH HYPOTHYROIDISM COMPARED WITH NORMAL SUBJECTS. PMID- 16694338 TI - THE RESPONSE OF THE GUINEA PIG'S RETICULOCYTES TO SUBSTANCES EFFECTIVE IN PERNICIOUS ANEMIA. A BIOLOGIC ASSAY OF THE THERAPEUTIC POTENCY OF LIVER EXTRACTS. PMID- 16694339 TI - THE ASSAY ON GUINEA PIGS OF THE HEMATOPOIETIC ACTIVITY OF HUMAN LIVERS, NORMAL AND PERNICIOUS ANEMIA. PMID- 16694340 TI - HETEROPHILE ANTIBODIES IN PNEUMONIA. PMID- 16694341 TI - THE METABOLIC EFFECTS OF HUMAN THYROGLOBULIN AND ITS PROTEOLYTIC CLEAVAGE PRODUCTS. PMID- 16694342 TI - A STUDY OF THE STANDARDIZATION OF DIGITALIS. I. A METHOD FOR CLINICAL STANDARDIZATION. PMID- 16694343 TI - A STUDY OF THE STANDARDIZATION OF DIGITALIS. II. THE RELATIONSHIP BETWEEN LABORATORY METHODS OF ASSAY AND POTENCY AS DETERMINED BY EXPERIMENTAL CUMULATIVE POISONING AND CLINICAL STANDARDIZATION. PMID- 16694344 TI - CHANGES IN THE BLOOD AND CIRCULATION WITH CHANGES IN POSTURE. THE EFFECT OF EXERCISE AND VASODILATATION. PMID- 16694345 TI - STUDIES ON THE IMMUNE RESPONSE OF THE RHEUMATIC SUBJECT AND ITS RELATIONSHIP TO ACTIVITY OF THE RHEUMATIC PROCESS. IV. CHARACTERISTICS OF STRAINS OF HEMOLYTIC STREPTOCOCCUS, EFFECTIVE AND NON-EFFECTIVE IN INITIATING RHEUMATIC ACTIVITY. PMID- 16694346 TI - STUDIES ON THE IMMUNE RESPONSE OF THE RHEUMATIC SUBJECT AND ITS RELATIONSHIP TO ACTIVITY OF THE RHEUMATIC PROCESS. V. ACTIVE AND PASSIVE IMMUNIZATION TO HEMOLYTIC STREPTOCOCCUS IN RELATION TO THE RHEUMATIC PROCESS. PMID- 16694347 TI - STUDIES ON THE IMMUNE RESPONSE OF THE RHEUMATIC SUBJECT AND ITS RELATIONSHIP TO ACTIVITY OF THE RHEUMATIC PROCESS. VI. THE SIGNIFICANCE OF THE RISE OF ANTISTREPTOLYSIN LEVEL IN THE DEVELOPMENT OF RHEUMATIC ACTIVITY. PMID- 16694348 TI - STUDIES ON THE IMMUNE RESPONSE OF THE RHEUMATIC SUBJECT AND ITS RELATIONSHIP TO ACTIVITY OF THE RHEUMATIC PROCESS. VII. SPLENECTOMY IN RELATION TO THE DEVELOPMENT OF RHEUMATIC ACTIVITY. PMID- 16694349 TI - A STUDY OF GASTRIC PEPSIN IN VARIOUS DISEASES. PMID- 16694350 TI - THE EFFECT OF SPLANCHNIC NERVE RESECTION AND SYMPATHETIC GANGLIONECTOMY IN A CASE OF PAROXYSMAL HEMOGLOBINURIA. PMID- 16694351 TI - ON THE NATURE OF THE SUBSTANCE(S) PRODUCING PAIN IN CONTRACTING SKELETAL MUSCLE: ITS BEARING ON THE PROBLEMS OF ANGINA PECTORIS AND INTERMITTENT CLAUDICATION. PMID- 16694352 TI - FURTHER OBSERVATIONS UPON THE CHANGES IN THE ELECTROLYTES OF THE URINE FOLLOWING THE INJECTION OF PARATHYROID EXTRACT. PMID- 16694353 TI - IMMUNIZATION OF HUMAN SUBJECTS WITH THE SPECIFIC CARBOHYDRATES OF TYPE III AND THE RELATED TYPE VIII PNEUMOCOCCUS. PMID- 16694354 TI - THE INFLUENCE OF DOSAGE AND ROUTE OF INJECTION ON THE ANTIBODY RESPONSE OF HUMAN SUBJECTS TO THE SPECIFIC CARBOHYDRATE OF THE TYPE VIII PNEUMOCOCCUS. PMID- 16694355 TI - COMPARISON OF HEMATOPOIESIS IN THE FETUS AND DURING RECOVERY FROM PERNICIOUS ANEMIA: TOGETHER WITH A CONSIDERATION OF THE RELATIONSHIP OF FETAL HEMATOPOIESIS TO MACROCYTIC ANEMIA OF PREGNANCY AND ANEMIA IN INFANTS. PMID- 16694356 TI - A CLINICAL AND EXPERIMENTAL STUDY OF THE STABILITY OF COLLOID OSMOTIC PRESSURE OF SERUM PROTEIN. PMID- 16694357 TI - THE RELATIONSHIP OF THE BLOOD GLUCOSE TO THE CONCENTRATION OF LACTOSE IN THE MILK OF LACTATING DIABETIC WOMEN. PMID- 16694358 TI - EXPERIMENTAL BUNDLE BRANCH BLOCK IN THE MONKEY. PMID- 16694359 TI - DIETARY PROTEIN IN HEMORRHAGIC BRIGHT'S DISEASE: II. The Effect of Diet on Serum Proteins, Proteinuria and Tissue Proteins. PMID- 16694360 TI - THE ADDIS SEDIMENT COUNT AND BLOOD UREA CLEARANCE TEST IN NORMAL PREGNANT WOMEN. PMID- 16694361 TI - INTUBATION STUDIES OF THE HUMAN SMALL INTESTINE. IV. CHEMICAL CHARACTERISTICS OF THE INTESTINAL CONTENTS IN THE FASTING STATE AND AS INFLUENCED BY THE ADMINISTRATION OF ACIDS, OF ALKALIES AND OF WATER. PMID- 16694363 TI - A MICRO METHOD FOR BLOOD UREA AND AN AUTOMATIC URINE COLLECTOR FOR UREA CLEARANCE IN INFANTS. PMID- 16694362 TI - STUDIES OF UREA EXCRETION. IX. COMPARISON OF UREA CLEARANCES CALCULATED FROM THE EXCRETION OF UREA, OF UREA PLUS AMMONIA, AND OF NITROGEN DETERMINABLE BY HYPOBROMITE. PMID- 16694364 TI - THE METABOLISM OF THE ISOLATED HEART OF DOGS RELATED TO AGE. PMID- 16694365 TI - STUDIES OF SODIUM AND POTASSIUM METABOLISM. THE EFFECT OF POTASSIUM ON THE SODIUM AND WATER BALANCES IN NORMAL SUBJECTS AND PATIENTS WITH BRIGHT'S DISEASE. PMID- 16694366 TI - STUDIES ON THE ANEMIA OF PELLAGRA. PMID- 16694367 TI - THE ACTION OF DINITROPHENOL AND INSULIN IN ACCELERATING THE METABOLISM OF ETHYL ALCOHOL. PMID- 16694368 TI - RADIATION OF HEAT FROM THE HUMAN BODY. V. THE TRANSMISSION OF INFRA-RED RADIATION THROUGH SKIN. PMID- 16694369 TI - PLASMA LIPIDS IN CHRONIC HEMORRHAGIC NEPHRITIS. PMID- 16694370 TI - PLASMA LIPIDS IN ESSENTIAL HYPERTENSION. PMID- 16694371 TI - THEOBROMINE SODIUM SALICYLATE AS A VASODILATOR. PMID- 16694372 TI - CHANGES OCCURRING IN THE MACROPHAGE SYSTEM OF THE LUNGS IN PNEUMOCOCCUS LOBAR PNEUMONIA. PMID- 16694373 TI - FILTERABLE SERUM CALCIUM IN LATE PREGNANT AND PARTURIENT WOMEN, AND IN THE NEWBORN. PMID- 16694374 TI - THE PROBLEM OF AN ANTIDIURETIC SUBSTANCE IN THE BLOOD OF PATIENTS WITH ECLAMPSIA AND OTHER HYPERTENSIVE DISEASES WITH OBSERVATIONS ON SPINAL FLUID. PMID- 16694375 TI - LIVER FUNCTION AS TESTED BY THE LIPEMIC CURVE AFTER INTRAVENOUS FAT ADMINISTRATION. PMID- 16694376 TI - STUDIES OF HEMOLYTIC STREPTOCOCCAL INFECTION I. FACTORS INFLUENCING THE OUTCOME OF ERYSIPELAS. PMID- 16694377 TI - STUDIES OF HEMOLYTIC STREPTOCOCCAL INFECTION II. THE SEROLOGICAL REACTIONS OF THE BLOOD DURING ERYSIPELAS. PMID- 16694378 TI - THE CIRCULATION RATE IN RELATION TO METABOLISM IN THYROID AND PITUITARY STATES (DECHOLIN METHOD). PMID- 16694379 TI - OBSERVATIONS ON ADRENAL INSUFFICIENCY. PMID- 16694380 TI - THE EFFECTS OF INHALATION OF HELIUM MIXED WITH OXYGEN ON THE MECHANICS OF RESPIRATION. PMID- 16694381 TI - THE NATURE OF THE PERIPHERAL RESISTANCE IN ARTERIAL HYPERTENSION WITH SPECIAL REFERENCE TO THE VASOMOTOR SYSTEM. PMID- 16694382 TI - THE EFFECT OF DRUGS IN THE PRODUCTION OF AGRANULOCYTOSIS WITH PARTICULAR REFERENCE TO AMIDOPYRINE HYPERSENSITIVITY. PMID- 16694383 TI - A TEST FOR ABNORMALLY LARGE AMOUNTS OF PARATHYROID HORMONE IN THE BLOOD. PMID- 16694384 TI - THE CAROTID SINUS REFLEX IN PATIENTS WITH HYPERTENSION. PMID- 16694385 TI - THE NUCLEOTIDE NITROGEN CONTENT OF PATHOLOGIC HUMAN WHOLE BLOOD. PMID- 16694386 TI - THE ROUTE OF INGESTED EGG WHITE TO THE SYSTEMIC CIRCULATION. PMID- 16694387 TI - THE SECRETION OF GASTRIC MUCIN IN MAN. A COMPARATIVE STUDY IN THE NORMAL SUBJECT AND IN THE PATIENT WITH PEPTIC ULCER IN RESPONSE TO AN ALCOHOL TEST MEAL. PMID- 16694388 TI - OBSERVATIONS ON THE NATURE OF THE SERUM PROTEINS IN NEPHROSIS. PMID- 16694389 TI - THE INFLUENCE OF HIGH AND LOW FAT DIETS AND THYROID SUBSTANCE ON PLASMA LIPIDS OF NEPHROTIC PATIENTS. PMID- 16694390 TI - A STUDY OF THE SERUM PHOSPHATASE IN BONE DISEASE. PMID- 16694391 TI - SERUM LIPOIDS IN MALNUTRITION. PMID- 16694392 TI - THE NATURE OF PLASMA AND URINARY PROTEINS IN NEPHROSIS. PMID- 16694393 TI - THE PHENOL RED CLEARANCE IN NORMAL MAN. PMID- 16694394 TI - A METHOD FOR MEASURING TONE AND REFLEX CONSTRICTION OF THE CAPILLARIES, VENULES AND VEINS OF THE HUMAN HAND WITH THE RESULTS IN NORMAL AND DISEASED STATES. PMID- 16694395 TI - STUDIES ON HYPOCHROMIC ANEMIA IN DOGS. I. THE EVALUATION OF A STANDARD BREAD DIET AND OF A MEAT DIET ON FORMATION OF HEMOGLOBIN BEFORE AND AFTER GASTRECTOMY. PMID- 16694396 TI - ADENOMA OF PANCREATIC ISLET CELLS WITH HYPOGLYCEMIA AND HYPERINSULINISM: Report of a Case with Studies on Blood Sugar and Metabolism before and after Operative Removal of Tumor. PMID- 16694397 TI - THE VOLUME OF THE EXTRACELLULAR FLUIDS OF THE BODY. PMID- 16694398 TI - STUDIES OF THE VARIATIONS IN THE ANTISTREPTOLYSIN TITER OF THE BLOOD SERUM FROM PATIENTS WITH HEMORRHAGIC NEPHRITIS. I. CONTROL OBSERVATIONS ON HEALTHY INDIVIDUALS AND PATIENTS SUFFERING FROM DISEASES OTHER THAN STREPTOCOCCAL INFECTIONS. PMID- 16694399 TI - STUDIES OF THE VARIATIONS IN THE ANTISTREPTOLYSIN TITER OF THE BLOOD SERUM FROM PATIENTS WITH HEMORRHAGIC NEPHRITIS. II. OBSERVATIONS ON PATIENTS SUFFERING FROM STREPTOCOCCAL INFECTIONS, RHEUMATIC FEVER AND ACUTE AND CHRONIC HEMORRHAGIC NEPHRITIS. PMID- 16694400 TI - THE EFFECT OF EPINEPHRIN ON THE BLOOD LIPOIDS OF NORMAL MAN. PMID- 16694401 TI - OBSERVATIONS ON THE FATE OF SODIUM SULFATE INJECTED INTRAVENOUSLY IN MAN. PMID- 16694402 TI - A COMPARISON OF THE ELECTROPHORETIC MOBILITIES AND SEDIMENTATION VELOCITIES OF RED CELLS FROM NORMAL AND PREGNANT HUMAN SUBJECTS. PMID- 16694403 TI - THE EFFECT OF THE TOXEMIAS OF PREGNANCY ON RENAL FUNCTION. PMID- 16694404 TI - PARATHYROID HORMONE IN THE BLOOD OF PREGNANT WOMEN. PMID- 16694405 TI - CONCERNING THE NATURALLY OCCURRING PORPHYRINS. IV. THE URINARY PORPHYRIN IN LEAD POISONING AS CONTRASTED WITH THAT EXCRETED NORMALLY AND IN OTHER DISEASES. PMID- 16694406 TI - THE NORMAL DURATION OF THE ELECTROCARDIOGRAPHIC VENTRICULAR COMPLEX. PMID- 16694407 TI - COMBINED CORD DEGENERATION WITHOUT ANEMIA: A CASE REPORT WITH STUDIES BEARING ON THE "INTRINSIC FACTOR" OF CASTLE. PMID- 16694408 TI - UNDERNUTRITION IN THE TREATMENT OF CORONARY ARTERY DISEASE (PARTICULARLY THROMBOSIS). EFFECT ON THE BASAL METABOLISM AND CIRCULATION. PMID- 16694409 TI - STUDIES OF ASCORBIC ACID AND RHEUMATIC FEVER I. QUANTITATIVE INDEX OF ASCORBIC ACID UTILIZATION IN HUMAN BEINGS AND ITS APPLICATION TO THE STUDY OF RHEUMATIC FEVER. PMID- 16694410 TI - STUDIES OF ASCORBIC ACID AND RHEUMATIC FEVER II. TEST OF PROPHYLACTIC AND THERAPEUTIC ACTION OF ASCORBIC ACID. PMID- 16694411 TI - A STUDY OF EXTERNAL PANCREATIC SECRETION IN MAN. PMID- 16694412 TI - SIMULTANEOUS PLASMA CLEARANCES OF CREATININE AND CERTAIN ORGANIC COMPOUNDS OF IODINE IN RELATION TO HUMAN KIDNEY FUNCTION. PMID- 16694413 TI - THE NITROGEN METABOLISM IN ANEMIA DURING THE REGENERATION OF BLOOD. PMID- 16694414 TI - METABOLIC STUDIES OF THE CHANGES IN BODY ELECTROLYTE AND DISTRIBUTION OF BODY WATER INDUCED EXPERIMENTALLY BY DEFICIT OF EXTRACELLULAR ELECTROLYTE. PMID- 16694415 TI - PASSIVE TRANSFER ANTIBODIES FOR SIX SAPROPHYTIC FUNGI IN A PATIENT WITH A SUPERFICIAL SCALING DERMATOSIS. PMID- 16694416 TI - CONCERNING THE SPECIFIC RESPONSE OF GUINEA PIG'S RETICULOCYTES TO SUBSTANCES EFFECTIVE IN PERNICIOUS ANEMIA. PMID- 16694417 TI - ACID-BASE EQUIVALENCE OF THE BLOOD IN DISEASES ASSOCIATED WITH HYPERGLOBULINEMIA; WITH SPECIAL REFERENCE TO LYMPHOGRANULOMA INGUINALE AND MULTIPLE MYELOMA. PMID- 16694418 TI - PLACENTAL INTERCHANGE. I. ON THE CONCENTRATION OF CERTAIN NITROGENOUS SUBSTANCES IN THE BLOOD, BEFORE AND AFTER PASSING THROUGH THE PLACENTA. PMID- 16694419 TI - THE FERROCYANIDE CLEARANCE IN MAN. PMID- 16694420 TI - PNEUMOCOCCIC INFECTIONS IN FAMILIES. PMID- 16694421 TI - BACTERIOLOGICAL AND IMMUNOLOGICAL STUDIES IN FAMILIES WITH PNEUMOCOCCIC INFECTIONS: THE DEVELOPMENT OF TYPE-SPECIFIC ANTIBODIES IN HEALTHY CONTACT CARRIERS. PMID- 16694422 TI - EXPERIMENTAL INDUCTION OF ERYTHEMA NODOSUM. PMID- 16694423 TI - VARIATIONS IN SERUM CALCIUM AND PHOSPHORUS DURING PREGNANCY. III. THE EFFECT ON THE FETAL CIRCULATION. PMID- 16694424 TI - VARIATIONS IN SERUM CALCIUM AND PHOSPHORUS DURING PREGNANCY. IV. EFFECT ON THE BODY STORES AS SHOWN BY THE ASH OF RATS. PMID- 16694425 TI - SUGARS AND GLYCOLYTIC ENZYMES OF SPINAL FLUID IN EPIDEMIC CEREBROSPINAL MENINGITIS. PMID- 16694426 TI - ENCEPHALITIS IN NORTH CHINA. RESULTS OBTAINED WITH NEUTRALIZATION TESTS. PMID- 16694427 TI - HEMOPHILIA. I. THE ABNORMAL COAGULATION OF THE BLOOD AND ITS RELATION TO THE BLOOD PLATELETS. PMID- 16694428 TI - EXPERIMENTAL HYPERTENSION-OBSERVATIONS ON SUSTAINED ELEVATION OF SYSTOLIC AND DIASTOLIC BLOOD PRESSURE IN DOGS. PMID- 16694429 TI - RETICULOCYTOSIS IN THE GUINEA PIG FOLLOWING INJECTIONS OF GASTRIC JUICE AND CONGO RED. PMID- 16694430 TI - THE COMPARISON OF UREA WITH UREA + AMMONIA CLEARANCES IN ACIDOTIC DOGS. PMID- 16694431 TI - RESPONSE TO INSULIN AS AN INDEX TO THE DIETARY MANAGEMENT OF DIABETES. PMID- 16694432 TI - THE HETEROPHILE ANTIBODIES IN INFECTIOUS MONONUCLEOSIS AND AFTER THE INJECTION OF SERUM. PMID- 16694433 TI - THE INFLUENCE OF CHANGES OF ABDOMINAL TENSION UPON PULMONARY FUNCTION. PMID- 16694434 TI - STUDIES ON THE CIRCULATION IN PREGNANCY. I. THE VELOCITY OF BLOOD FLOW AND RELATED ASPECTS OF THE CIRCULATION IN NORMAL PREGNANT WOMEN. PMID- 16694435 TI - THE BLOOD LIPIDS OF DIABETIC CHILDREN. PMID- 16694436 TI - HEMATOLOGICAL STUDIES IN HYPOTHYROIDISM FOLLOWING TOTAL THYROIDECTOMY. PMID- 16694437 TI - THE WORK OF THE LEFT VENTRICLE IN AORTIC INSUFFICIENCY. PMID- 16694438 TI - THE PERIPHERAL BLOOD FLOW IN SURGICAL SHOCK: The Reduction in Circulation through the Hand Resulting from Pain, Fear, Cold, and Asphyxia, with Quantitative Measurements of the Volume Flow of Blood in Clinical Cases of Surgical Shock. PMID- 16694439 TI - THE GLYCEMIC RESPONSE TO ISOGLUCOGENIC QUANTITIES OF PROTEIN AND CARBOHYDRATE. PMID- 16694440 TI - THE ADVANTAGE OF A HIGH PROTEIN DIET IN THE TREATMENT OF SPONTANEOUS HYPOGLYCEMIA: Preliminary Report. PMID- 16694441 TI - THE METABOLISM OF HUMAN ERYTHROBLASTS. PMID- 16694442 TI - THE SALT AND WATER METABOLISM OF ADRENAL INSUFFICIENCY AND PARTIAL STARVATION IN RATS. PMID- 16694443 TI - INTRACELLULAR FLUID LOSS IN HEMORRHAGE. PMID- 16694444 TI - THE EFFECT OF DIETARY PROTEIN ON THE UREA CLEARANCE OF CHILDREN WITH NEPHROSIS. PMID- 16694445 TI - THERAPEUTIC SERUM FOR PNEUMOCOCCUS TYPE V (COOPER) PNEUMONIA. PMID- 16694446 TI - OBSERVATIONS ON THE DEVELOPMENT OF THE HIGH BLOOD SEDIMENTATION RATE IN RHEUMATIC CARDITIS. PMID- 16694447 TI - DISTRIBUTION OF RESPIRATORY GASES IN A CLOSED BREATHING CIRCUIT. I. IN NORMAL SUBJECTS. PMID- 16694448 TI - HYPERPARATHYROIDISM IN KIDNEY DISEASE. PMID- 16694449 TI - STUDIES ON HYPOCHROMIC ANEMIA IN DOGS. II. THE EVALUATION OF PREDIGESTED BEEF, IRON AND LIVER EXTRACT ON FORMATION OF HEMOGLOBIN AFTER GASTRECTOMY. PMID- 16694450 TI - HEMOPHILIA. II. SOME PROPERTIES OF A SUBSTANCE OBTAINED FROM NORMAL HUMAN PLASMA EFFECTIVE IN ACCELERATING THE COAGULATION OF HEMOPHILIC BLOOD. PMID- 16694451 TI - THE ARTIFICIAL INDUCTION OF SUBCUTANEOUS NODULES IN PATIENTS WITH RHEUMATIC FEVER. PMID- 16694452 TI - THE PATHOLOGY OF SPONTANEOUS AND INDUCED SUBCUTANEOUS NODULES IN RHEUMATIC FEVER. PMID- 16694453 TI - DEVELOPMENT OF ANTIFIBRINOLYTIC PROPERTIES IN BLOOD OF PATIENTS WITH RHEUMATIC FEVER, CHRONIC INFECTIVE ARTHRITIS AND BACTERIAL ENDOCARDITIS. PMID- 16694454 TI - COMPLEMENT FIXATION TESTS IN PERTUSSIS. PMID- 16694456 TI - METABOLIC STUDIES ON CHRONIC ULCERATIVE COLITIS. PMID- 16694455 TI - STUDIES OF HEMOLYTIC STREPTOCOCCAL INFECTION. III. THE CHARACTERISTICS OF THE HEMOLYTIC STREPTOCOCCI ISOLATED FROM PATIENTS WITH ERYSIPELAS. PMID- 16694457 TI - RESPIRATORY RESPONSE DURING EXERCISE IN PULMONARY FIBROSIS AND EMPHYSEMA. PMID- 16694458 TI - A STUDY OF THE CARRIER CONDITION ASSOCIATED WITH TYPE II PNEUMONIA IN A CAMP OF THE CIVILIAN CONSERVATION CORPS. PMID- 16694459 TI - THE EFFECT ON THE KIDNEY OF BILATERAL SPLANCHNICECTOMY IN PATIENTS WITH HYPERTENSION. PMID- 16694460 TI - STUDIES OF GALLBLADDER FUNCTION. XV. CHOLESTEROL IN HUMAN LIVER BILE. PMID- 16694461 TI - THE NATURE OF CIRCULATORY COLLAPSE INDUCED BY SODIUM NITRITE. PMID- 16694462 TI - THE ROLE OF THE VENOUS SYSTEM IN CIRCULATORY COLLAPSE INDUCED BY SODIUM NITRITE. PMID- 16694463 TI - DISTRIBUTION OF RESPIRATORY GASES IN A CLOSED BREATHING CIRCUIT. II. PULMONARY FIBROSIS AND EMPHYSEMA. PMID- 16694464 TI - THE OSMOTIC PRESSURE OF PROTEINS IN WHOLE SERUM. PMID- 16694465 TI - STUDIES OF GONOCOCCAL INFECTION. I. A STUDY OF THE MODE OF DESTRUCTION OF THE GONOCOCCUS IN VITRO. PMID- 16694466 TI - STUDIES OF GONOCOCCAL INFECTION. II. THE BACTERIOLYTIC POWER OF THE WHOLE DEFIBRINATED BLOOD OF PATIENTS WITH GONOCOCCAL ARTHRITIS. PMID- 16694467 TI - THE CLINICAL USES OF HUMAN SERUMS PRESERVED BY THE LYOPHILE PROCESS. PMID- 16694468 TI - THE RESPONSE OF NORMAL INDIVIDUALS AND PATIENTS WITH DIABETES INSIPIDUS TO THE INGESTION OF WATER. PMID- 16694470 TI - PROGNOSTIC VALUE OF THE PRECIPITIN TEST IN MENINGOCOCCUS MENINGITIS. PMID- 16694469 TI - PARATHYROID HYPERPLASIA IN RABBITS PRODUCED BY PARENTERAL PHOSPHATE ADMINISTRATION. PMID- 16694471 TI - THERMAL INJURIES: THE EFFECTS OF FREEZING. PMID- 16694472 TI - THE ABSORPTION AND EXCRETION OF CALCIUM AND PHOSPHORUS IN THREE PATIENTS WITH COLOSTOMY AND ILEOSTOMY. PMID- 16694473 TI - THE GLYCINE SYNTHESIS IN PATIENTS WITH PROGRESSIVE MUSCULAR DYSTROPHY. PMID- 16694474 TI - RESULTS OF IMMUNIZATION BY MEANS OF ACTIVE VIRUS OF HUMAN INFLUENZA. PMID- 16694475 TI - THE ABSORPTION OF HEXOSES FROM THE UPPER PART OF THE SMALL INTESTINE IN MAN. PMID- 16694476 TI - GALACTOSE TOLERANCE IN HYPERTHYROIDISM. PMID- 16694477 TI - STUDIES OF AN URTICARIAL RESPONSE TO BLUE AND VIOLET LIGHT IN MAN. PMID- 16694478 TI - VARIATIONS IN THE PERMEABILITY OF THE RED BLOOD CELLS IN MAN, WITH PARTICULAR REFERENCE TO THE CONDITIONS OBTAINING IN PERNICIOUS ANEMIA. PMID- 16694479 TI - THE INFLUENCE OF MUSCULAR EXERCISE ON BLOOD SUGAR CONCENTRATIONS. PMID- 16694480 TI - CLINICAL STUDIES OF THE BLOOD VOLUME. I. CLINICAL APPLICATION OF A METHOD EMPLOYING THE AZO DYE "EVANS BLUE" AND THE SPECTROPHOTOMETER. PMID- 16694481 TI - CLINICAL STUDIES OF THE BLOOD VOLUME. II. THE RELATION OF PLASMA AND TOTAL BLOOD VOLUME TO VENOUS PRESSURE, BLOOD VELOCITY RATE, PHYSICAL MEASUREMENTS, AGE AND SEX IN NINETY NORMAL HUMANS. PMID- 16694482 TI - RELATION OF THYROID AND PARATHYROID GLANDS TO CALCIUM AND PHOSPHORUS METABOLISM. STUDY OF A CASE WITH COEXISTENT HYPOPARATHYROIDISM AND HYPERTHYROIDISM. PMID- 16694483 TI - GASTRO-INTESTINAL STUDIES. VII. THE EXCRETION OF XYLOSE IN PERNICIOUS ANEMIA. PMID- 16694484 TI - THE SIGNIFICANCE OF THE SODIUM AND POTASSIUM CONTENT OF MUSCLE TISSUE AND THE RELATION OF THE AMOUNT OF EDEMA FLUID IN MUSCLE TO THE LEVEL OF SERUM PROTEIN IN EXPERIMENTAL NUTRITIONAL EDEMA. PMID- 16694486 TI - CONCERNING THE NATURALLY OCCURRING PORPHYRINS. V. PORPHYRINS OF THE FECES. PMID- 16694485 TI - THE COMPOSITION OF HUMAN GALLBLADDER BILE AND ITS RELATIONSHIP TO CHOLELITHIASIS. PMID- 16694487 TI - PATHOLOGY OF PREGNANCY TOXEMIAS. PMID- 16694488 TI - RENAL PHYSIOLOGY IN LOBAR PNEUMONIA. PMID- 16694489 TI - STUDIES OF THE CIRCULATION IN PERNICIOUS ANEMIA. PMID- 16694490 TI - DETERMINATION OF BLOOD ASCORBIC ACID. PMID- 16694491 TI - NEUTRALIZATION TESTS IN POLIOMYELITIS. SERA TAKEN DURING THE ACUTE AND CONVALESCENT STAGES OF THE DISEASE AND TESTED WITH A PASSAGE VIRUS AND A STRAIN ISOLATED DURING THE 1935 NEW YORK CITY OUTBREAK. PMID- 16694492 TI - MACROCYTIC ANEMIA IN PREGNANT WOMEN WITH VITAMIN B DEFICIENCY. PMID- 16694493 TI - CHRONAXIMETRIC EXAMINATIONS IN B AVITAMINOSIS DURING PREGNANCY. PMID- 16694494 TI - THE EFFECT OF HEATING WITH ALKALI ON THE CALORIGENIC ACTIVITY OF DESICCATED THYROID AND OF THYROXINE. PMID- 16694495 TI - MAGNESIUM METABOLISM IN HEALTH AND DISEASE. I. THE MAGNESIUM AND CALCIUM EXCRETION OF NORMAL INDIVIDUALS, ALSO THE EFFECTS OF MAGNESIUM, CHLORIDE, AND PHOSPHATE IONS. PMID- 16694496 TI - MAGNESIUM METABOLISM IN HEALTH AND DISEASE. II. THE EFFECT OF THE PARATHYROID HORMONE. PMID- 16694497 TI - MAGNESIUM METABOLISM IN HEALTH AND DISEASE. III. IN EXOPHTHALMIC GOITER, BASOPHILIC ADENOMA, ADDISON'S DISEASE AND STEATORRHEA. PMID- 16694498 TI - STUDIES IN TEMPERATURE SENSATION. I. A COMPARISON OF THE SENSATION PRODUCED BY INFRA-RED AND VISIBLE RADIATION. PMID- 16694499 TI - STUDIES IN TEMPERATURE SENSATION. II. THE TEMPERATURE CHANGES RESPONSIBLE FOR THE STIMULATION OF THE HEAT END ORGANS. PMID- 16694500 TI - STUDIES IN TEMPERATURE SENSATION. III. THE SENSITIVITY OF THE BODY TO HEAT AND THE SPATIAL SUMMATION OF THE END ORGAN RESPONSES. PMID- 16694501 TI - ACUTE MOUNTAIN SICKNESS; THE EFFECT OF AMMONIUM CHLORIDE. PMID- 16694502 TI - THE USE OF FERROUS GLUCONATE IN THE TREATMENT OF HYPOCHROMIC ANEMIA. PMID- 16694503 TI - RHEUMATIC FEVER AS A FAMILIAL DISEASE. ENVIRONMENT, COMMUNICABILITY AND HEREDITY IN THEIR RELATION TO THE OBSERVED FAMILIAL INCIDENCE OF THE DISEASE. PMID- 16694505 TI - STUDIES OF THE PRINCIPLE IN LIVER EFFECTIVE IN PERNICIOUS ANEMIA. IV. THE THERAPEUTIC ACTIVITY OF ITS MULTIPLE FACTORS. PMID- 16694504 TI - SPLENIC VEIN PRESSURE IN CONGESTIVE SPLENOMEGALY (BANTI'S SYNDROME). PMID- 16694506 TI - VITAMIN C SATURATION LEVELS IN THE BODY IN NORMAL SUBJECTS AND IN VARIOUS PATHOLOGICAL CONDITIONS. PMID- 16694507 TI - STUDIES ON THE ELASTIC PROPERTIES OF HUMAN ISOLATED AORTA. PMID- 16694508 TI - CALCIUM AND PHOSPHORUS METABOLISM IN OSTEOMALACIA. V. THE EFFECT OF VARYING LEVELS AND RATIOS OF CALCIUM TO PHOSPHORUS INTAKE ON THEIR SERUM LEVELS, PATHS OF EXCRETION AND BALANCES, IN THE PRESENCE OF CONTINUOUS VITAMIN D THERAPY. PMID- 16694509 TI - STUDIES IN IRON TRANSPORTATION AND METABOLISM. I. CHEMICAL METHODS AND NORMAL VALUES FOR PLASMA IRON AND "EASILY SPLIT-OFF" BLOOD IRON. PMID- 16694510 TI - STUDIES IN IRON TRANSPORTATION AND METABOLISM. II. THE MECHANISM OF IRON TRANSPORTATION: ITS SIGNIFICANCE IN IRON UTILIZATION IN ANEMIC STATES OF VARIED ETIOLOGY. PMID- 16694511 TI - THE RESULT OF INTRA-ARTERIAL INJECTION OF VASODILATING DRUGS ON THE CIRCULATION: OBSERVATIONS ON VASOMOTOR GRADIENT. PMID- 16694512 TI - THE VALIDITY OF THE CALCULATION OF STANDARD UREA CLEARANCES FROM LOW URINE VOLUMES. PMID- 16694513 TI - CHANGES IN THE VASOMOTOR REACTION ASSOCIATED WITH GLOMUS TUMORS. PMID- 16694514 TI - THE DAILY URINARY EXCRETION OF ESTROGENIC AND ANDROGENIC SUBSTANCES BY NORMAL MEN AND WOMEN. PMID- 16694515 TI - THE URINARY EXCRETION OF ANDROGENIC AND ESTROGENIC SUBSTANCES IN CERTAIN ENDOCRINE STATES. STUDIES IN HYPOGONADISM, GYNECOMASTIA AND VIRILISM. PMID- 16694516 TI - NUTRITIONAL EDEMA IN THE DOG. V. DEVELOPMENT OF DEFICITS IN ERYTHROCYTES AND HEMOGLOBIN ON A DIET DEFICIENT IN PROTEIN. PMID- 16694517 TI - LOSS OF MINERALS THROUGH THE SKIN OF NORMAL HUMANS WHEN SWEATING IS AVOIDED. PMID- 16694518 TI - ADJUSTMENT OF THE FLOW OF TISSUE FLUID IN THE PRESENCE OF LOCALIZED, SUSTAINED HIGH VENOUS PRESSURE AS FOUND WITH VARICES OF THE GREAT SAPHENOUS SYSTEM DURING WALKING. PMID- 16694519 TI - THE COAGULATION DEFECT IN HEMOPHILIA. THE EFFECT IN HEMOPHILIA OF INTRAMUSCULAR ADMINISTRATION. OF A GLOBULIN SUBSTANCE DERIVED FROM NORMAL HUMAN PLASMA. PMID- 16694520 TI - THE OPSONO-CYTOPHAGIC TEST IN CHILDREN WITH PERTUSSIS AND IN CHILDREN VACCINATED WITH H. PERTUSSIS ANTIGENS. PMID- 16694521 TI - PLASMA CHOLESTEROL SATURATION IN PATIENTS WITH HYPERTENSION. WITH A NOTE ON PREPARATION OF GLASS FILTERS FOR MICRO-FILTRATION OF CHOLESTEROL DIGITONIDE. PMID- 16694523 TI - STUDIES IN THE PHYSIOLOGY OF BLOOD VESSELS IN MAN. APPARATUS AND METHODS. I. A SENSITIVE PLETHYS-MOSPHYGMOGRAPH FOR A PORTION OF THE FINGER. PMID- 16694522 TI - STUDIES ON THE MECHANISM OF PROTEINURIA. PMID- 16694524 TI - STUDIES IN THE PHYSIOLOGY OF BLOOD VESSELS IN MAN. APPARATUS AND METHODS. II. A METHOD FOR THE DETERMINATION OF THE VOLUME OF THE SOFT TISSUE ABOUT THE TERMINAL PHALANX OF THE HUMAN FINGER. PMID- 16694525 TI - STUDIES IN THE PHYSIOLOGY OF BLOOD VESSELS IN MAN. III. SOME EFFECTS OF RAISING AND LOWERING THE ARM UPON THE PULSE VOLUME AND BLOOD VOLUME OF THE HUMAN FINGER TIP IN HEALTH AND IN CERTAIN DISEASES OF THE BLOOD VESSELS. PMID- 16694527 TI - THE OPSONO-CYTOPHAGIC REACTION OF THE BLOOD IN PERTUSSIS. PMID- 16694526 TI - A CLINICAL STUDY OF THE ACTION OF 10 COMMONLY USED DRUGS ON CARDIAC OUTPUT, WORK AND SIZE; ON RESPIRATION, ON METABOLIC RATE AND ON THE ELECTROCARDIOGRAM. PMID- 16694528 TI - THE EFFECT OF IMMUNE BLOOD UPON THE OPSONO-CYTOPHAGIC POWER OF THE BLOOD IN PERTUSSIS. PMID- 16694529 TI - THE BISULPHITE BINDING POWER OF THE BLOOD IN HEALTH AND IN DISEASE, WITH SPECIAL REFERENCE TO VITAMIN B(1) DEFICIENCY. PMID- 16694530 TI - THE ESTIMATION OF THE SUBCUTANEOUS TISSUE PRESSURE BY A DIRECT METHOD. PMID- 16694531 TI - CLINICAL STUDIES OF THE BLOOD VOLUME. III. CHANGES IN BLOOD VOLUME, VENOUS PRESSURE AND BLOOD VELOCITY RATE IN CHRONIC CONGESTIVE HEART FAILURE. PMID- 16694532 TI - THE MEASUREMENT OF GLOMERULAR FILTRATION. CREATININE, SUCROSE AND UREA CLEARANCES IN SUBJECTS WITHOUT RENAL DISEASE. PMID- 16694533 TI - THE MEASUREMENT OF GLOMERULAR FILTRATION. THE CREATININE, SUCROSE AND UREA CLEARANCES IN SUBJECTS WITH RENAL DISEASE. PMID- 16694534 TI - A CONVENIENT METHOD FOR THE DETERMINATION OF THE APPROXIMATE CARDIAC OUTPUT IN MAN. PMID- 16694535 TI - CHRONIC PYELONEPHRITIS AND ARTERIAL HYPERTENSION. PMID- 16694536 TI - ALLERGY AND DESENSITIZATION IN TUBERCULOSIS. PMID- 16694537 TI - RELATION OF SERUM CALCIUM TO SERUM ALBUMIN AND GLOBULINS. PMID- 16694538 TI - STUDIES ON SERUM PROTEINS. I. IDENTIFICATION OF A SINGLE SERUM GLOBULIN BY IMMUNOLOGICAL MEANS. ITS DISTRIBUTION IN THE SERA OF NORMAL INDIVIDUALS AND OF PATIENTS WITH CIRRHOSIS OF THE LIVER AND WITH CHRONIC GLOMERULONEPHRITIS. PMID- 16694539 TI - A QUANTITATIVE STUDY OF THE OXIDATION OF GLUCOSE IN NORMAL AND DIABETIC MEN. PMID- 16694540 TI - CHLORIDE DEPLETION IN CONDITIONS OTHER THAN ADDISON'S DISEASE. PMID- 16694541 TI - THE EXCRETION OF COPROPORPHYRIN I FOLLOWING HEMORRHAGE IN DOGS. PMID- 16694542 TI - THE INFLUENCE OF EPINEPHRINE ON THE DIGITAL ARTERIOLES OF MAN: A STUDY OF THE VASOCONSTRICTOR EFFECTS. PMID- 16694543 TI - STUDIES OF GONOCOCCAL INFECTION. III. A COMPARISON OF THE BACTERICIDAL PROPERTIES OF THE SYNOVIAL FLUID AND BLOOD IN GONOCOCCAL ARTHRITIS. PMID- 16694544 TI - STUDIES OF GONOCOCCAL INFECTION. IV. THE EFFECT OF MUCIN ON THE BACTERIOLYTIC POWER OF WHOLE BLOOD AND IMMUNE SERUM. PMID- 16694545 TI - THE RENAL EXCRETION OF ENDOGENOUS CREATININE IN MAN. COMPARISON WITH EXOGENOUS CREATININE AND INULIN. PMID- 16694546 TI - THE EFFECT OF EPINEPHRIN IN CIRCULATORY COLLAPSE INDUCED BY SODIUM NITRITE. PMID- 16694547 TI - THE NEUTRALIZATION OF ENCEPHALITIS VIRUS (ST. LOUIS, 1933), BY SERUM. PMID- 16694548 TI - RESPIRATORY PROPERTIES OF THE ARTERIAL BLOOD IN NORMAL MAN AND IN PATIENTS WITH DISEASE OF THE LIVER: POSITION OF THE OXYGEN DISSOCIATION CURVE. PMID- 16694549 TI - OBSERVATIONS ON THE RENAL THRESHOLD FOR ASCORBIC ACID IN MAN. PMID- 16694550 TI - THE EFFECTS ON THE CARDIOVASCULAR SYSTEM OF FLUIDS ADMINISTERED INTRAVENOUSLY IN MAN. I. STUDIES OF THE AMOUNT AND DURATION OF CHANGES IN BLOOD VOLUME. PMID- 16694551 TI - THE DISTRIBUTION OF BODY WATER AND ELECTROLYTES IN ADRENAL INSUFFICIENCY. PMID- 16694552 TI - THE EFFECT OF HYPERTHERMIA ON THE DISTRIBUTION OF WATER AND ELECTROLYTES IN BRAIN, MUSCLE AND LIVER. PMID- 16694553 TI - THE METABOLISM OF BLOOD PIGMENTS IN PERNICIOUS ANEMIA. PMID- 16694554 TI - UREA EXCRETION AT LOW URINE VOLUMES. THE CALCULATION OF "MINIMAL" UREA CLEARANCES. PMID- 16694555 TI - THE EXCRETION OF PORPHYRIN IN REFRACTORY AND APLASTIC ANEMIA. PMID- 16694556 TI - THE EFFECT OF ARTIFICIAL FEVER AND SPECIFIC ANTISERUM ON THE ORGANISMS PRESENT IN CASES OF TYPE III PNEUMOCOCCUS MENINGITIS. PMID- 16694557 TI - THE FLOW OF BLOOD IN THE CORONARY ARTERIES IN PATHOLOGICAL HEARTS. PMID- 16694558 TI - CLINICAL STUDIES OF THE BLOOD VOLUME. IV. ADAPTATION OF THE METHOD TO THE PHOTOELECTRIC MICROCOLORIMETER. PMID- 16694559 TI - ALKALINE TIDES. PMID- 16694560 TI - A NOTE ON THE STATE OF CALCIUM IN HIGH PROTEIN SERUM. PMID- 16694561 TI - ASCORBIC ACID IN CEREBROSPINAL FLUID. PMID- 16694562 TI - THE CONCENTRATION OF GLUCOSAMINE IN NORMAL AND PATHOLOGICAL SERA. PMID- 16694563 TI - A STUDY OF THE BLOOD IODINE IN CHILDHOOD. PMID- 16694564 TI - THE EFFECTS OF PRESSOR DRUGS AND OF SALINE KIDNEY EXTRACTS ON BLOOD PRESSURE AND SKIN TEMPERATURE. PMID- 16694565 TI - THE EFFECT OF ECLAMPTIC BLOOD UPON THE URINARY OUTPUT AND BLOOD PRESSURE OF HUMAN RECIPIENTS. PMID- 16694567 TI - EFFECT OF INSULIN ON THE CONCENTRATION OF URIC ACID IN THE BLOOD. PMID- 16694566 TI - STUDIES IN THE PHYSIOLOGY OF ARTIFICIAL FEVER. I. CHANGES IN THE BLOOD VOLUME AND WATER BALANCE. PMID- 16694568 TI - STUDIES OF THE CIRCULATION IN PATIENTS SUFFERING FROM SPONTANEOUS MYXEDEMA. PMID- 16694569 TI - HEAT STROKE: CLINICAL AND CHEMICAL OBSERVATIONS ON 44 CASES. PMID- 16694570 TI - THE MEASUREMENT OF THE TUBULAR EXCRETORY MASS, EFFECTIVE BLOOD FLOW AND FILTRATION RATE IN THE NORMAL HUMAN KIDNEY. PMID- 16694571 TI - THE EFFECT OF ALCOHOL ON THE WATER AND ELECTROLYTE BALANCE IN MAN. PMID- 16694572 TI - ESTIMATIONS OF THE WORK OF THE HEART DURING AND BETWEEN ATTACKS OF ANGINA PECTORIS. PMID- 16694573 TI - CRITICAL REMARKS ON THE DETERMINATION OF URINARY EXCRETION OF ASCORBIC ACID. PMID- 16694574 TI - THE GUANIDINE BASES IN THE BLOOD OF DOGS WITH EXPERIMENTAL HYPERTENSION PRODUCED BY CONSTRICTION OF THE RENAL ARTERIES. PMID- 16694575 TI - THE EFFECT OF VITAMIN D ON CALCIUM AND PHOSPHORUS METABOLISM; STUDIES ON FOUR PATIENTS. PMID- 16694576 TI - A COMPARISON OF THE EFFECTS OF A.T. 10 (DIHYDROTACHYSTEROL) AND VITAMIN D ON CALCIUM AND PHOSPHORUS METABOLISM IN HYPOPARATHYROIDISM. PMID- 16694577 TI - EVAPORATION OF BODY WATER IN LOBAR PNEUMONIA. PMID- 16694578 TI - OBSERVATIONS ON THE ETIOLOGIC RELATIONSHIP OF ACHYLIA GASTRICA TO PERNICIOUS ANEMIA. VII. RESEMBLANCES BETWEEN THE PROTEOLYTIC ACTIVITY OF NORMAL HUMAN GASTRIC JUICE ON CASEIN IN NEUTRAL SOLUTION AND THE ACTIVITY OF THE INTRINSIC FACTOR. PMID- 16694579 TI - THE EXCRETION OF UREA IN NORMAL MAN AND IN SUBJECTS WITH GLOMERULONEPHRITIS. PMID- 16694580 TI - THE EFFECT OF TOTAL SYMPATHECTOMY ON THE OCCURRENCE OF SHOCK FROM HEMORRHAGE. PMID- 16694581 TI - A SIMPLE METHOD FOR THE ESTIMATION OF TOTAL PROTEIN CONTENT OF PLASMA AND SERUM. I. A FALLING DROP METHOD FOR THE DETERMINATION OF SPECIFIC GRAVITY. PMID- 16694582 TI - A SIMPLE METHOD FOR THE ESTIMATION OF TOTAL PROTEIN CONTENT OF PLASMA AND SERUM. II. THE ESTIMATION OF TOTAL PROTEIN CONTENT OF HUMAN PLASMA AND SERUM BY THE USE OF THE FALLING DROP METHOD. PMID- 16694583 TI - THE CHEMICAL COMPOSITION OF VOLUNTARY MUSCLE IN MUSCLE DISEASE: A COMPARISON OF PROGRESSIVE MUSCULAR DYSTROPHY WITH OTHER DISEASES TOGETHER WITH A STUDY OF EFFECTS OF GLYCINE AND CREATINE THERAPY. PMID- 16694584 TI - THE LATE EFFECTS OF BILATERAL CAROTID SINUS DENERVATION IN MAN: Report of Two Cases with Studies of the Vascular Reflexes. PMID- 16694585 TI - THE RENAL FACTOR IN ARTERIAL HYPERTENSION WITH COARCTATION OF THE AORTA. PMID- 16694586 TI - THE EFFECTS ON THE CARDIOVASCULAR SYSTEM OF FLUIDS ADMINISTERED INTRAVENOUSLY IN MAN. II. THE DYNAMICS OF THE CIRCULATION. PMID- 16694587 TI - CHANGES IN BLOOD AND INTERSTITIAL FLUID RESULTING FROM SURGICAL OPERATION AND ETHER ANESTHESIA. PMID- 16694588 TI - OBSERVATIONS ON THE BLOOD OF WORKMEN EXPOSED TO HIGH TEMPERATURES. PMID- 16694589 TI - THE NATURE OF THE LOWERED RESISTANCE TO INFECTION IN DIABETES MELLITUS. PMID- 16694590 TI - ON THE ACTIONS OF CRYSTALLINE VITAMIN D(2) (CALCIFEROL) IN CHRONIC PARATHYROID TETANY. PMID- 16694591 TI - CYANOSIS WITHOUT SULF- OR METHEMOGLOBINEMIA IN PATIENTS RECEIVING SULFANILAMIDE TREATMENT. PMID- 16694592 TI - STUDIES OF THE CIRCULATION IN THE PRESENCE OF ABNORMAL CARDIAC RHYTHMS. OBSERVATIONS RELATING TO (PART I) RHYTHMS ASSOCIATED WITH RAPID VEN-TRICULAR RATE AND TO (PART II) RHYTHMS ASSOCIATED WITH SLOW VENTRICULAR RATE. PMID- 16694593 TI - A STUDY OF SOME OF THE PHYSIOLOGICAL EFFECTS OF SULFANILAMIDE. I. CHANGES IN THE ACID BASE BALANCE. PMID- 16694594 TI - AN " ACID " PHOSPHATASE OCCURRING IN THE SERUM OF PATIENTS WITH METASTASIZING CARCINOMA OF THE PROSTATE GLAND. PMID- 16694595 TI - REACTIONS OF HUMAN SUBJECTS TO THE INJECTION OF PURIFIED TYPE SPECIFIC PNEUMOCOCCUS POLYSACCHARIDES. PMID- 16694597 TI - PRESIDENT'S ADDRESS CLINICAL EPIDEMIOLOGY. PMID- 16694596 TI - TISSUE PRESSURE (INTRACUTANEOUS, SUBCUTANEOUS, AND INTRAMUSCULAR) AS RELATED TO VENOUS PRESSURE, CAPILLARY FILTRATION, AND OTHER FACTORS. PMID- 16694598 TI - CALCIUM AND PHOSPHORUS METABOLISM IN DISEASES OF THE THYROPARATHYROID APPARATUS. II. CALCIUM AND PHOSPHORUS BALANCE (A) FOLLOWING THERAPEUTIC RADIATION OF THE HYPERPLASTIC THYROID GLAND, AND (B) IN HYPERTHYROIDIC PATIENTS TREATED WITH IODINE. PMID- 16694599 TI - STUDY OF THE TENDENCY TO EDEMA FORMATION ASSOCIATED WITH INCOMPETENCE OF THE VALVES OF THE COMMUNICATING VEINS OF THE LEG. OXYGEN TENSION OF THE BLOOD CONTAINED IN VARICOSE VEINS. PMID- 16694600 TI - THE DETERMINATION OF THE CARDIAC OUTPUT IN MAN AT BRIEF INTERVALS BY A MODIFICATION OF THE ETHYL IODIDE METHOD. PMID- 16694601 TI - THE CARDIAC OUTPUT AND OXYGEN CONSUMPTION OF NINE SURGICAL PATIENTS BEFORE AND AFTER OPERATION. PMID- 16694602 TI - MEASUREMENTS OF THE CIRCULATION IN CONSTRICTIVE PERICARDITIS BEFORE AND AFTER RESECTION OF THE PERICARDIUM. PMID- 16694603 TI - RENAL EXCRETION AT LOW URINE VOLUMES AND THE MECHANISM OF OLIGURIA. PMID- 16694604 TI - THE VALUE OF THE ACID TEST MEAL: A STUDY OF NORMAL PERSONS AND OF PERSONS WITH DUODENAL ULCER. PMID- 16694605 TI - PLACENTAL INTERCHANGE. II. COMPARISON OF THE TOTAL BASE CONCENTRATION OF THE FETAL AND MATERNAL BLOOD AT PARTURITION. PMID- 16694606 TI - AN ATTEMPT TO INCREASE RESISTANCE TO PERTUSSIS IN NEWBORN INFANTS BY IMMUNIZING THEIR MOTHERS DURING PREGNANCY. PMID- 16694607 TI - ON THE PERMANENCE OF RECOVERY IN ACUTE GLOMERULONEPHRITIS. PMID- 16694608 TI - THE SERUM ANTISTREPTOLYSIN TITER IN ACUTE GLOMERULONEPHRITIS. PMID- 16694609 TI - EXPERIENCE WITH THE HAMILTON AND HIGHMAN TEST FOR PARATHYROID HYPERFUNCTION IN CHRONIC NEPHRITIS, TOXIC GOITER, AND PAGET'S DISEASE OF BONE. PMID- 16694610 TI - THE CHOLINE-ESTERASE ACTIVITY OF THE BLOOD SERUM IN DISEASE. PMID- 16694612 TI - BASAL GASTRIC SECRETION IN CASES OF PEPTIC ULCER: RELATION OF ACIDITY TO HEALING OF ULCER. PMID- 16694611 TI - THE EFFECT OF ARTIFICIAL PNEUMOTHORAX UPON THE ANOXEMIA OF PNEUMONIA. PMID- 16694613 TI - REQUIREMENTS FOR VITAMIN C IN MAN. PMID- 16694614 TI - THE USE OF A GLOBULIN SUBSTANCE DERIVED FROM BEEF PLASMA AS A LOCAL HEMOSTATIC IN HEMOPHILIA. PMID- 16694615 TI - THE CONTROL OF RENAL BLOOD FLOW AND GLOMERULAR FILTRATION IN NORMAL MAN. PMID- 16694617 TI - A PLETHYSMOGRAPHIC METHOD FOR THE QUANTITATIVE MEASUREMENT OF THE BLOOD FLOW IN THE FOOT. PMID- 16694616 TI - A STUDY OF SOME OF THE PHYSIOLOGICAL EFFECTS OF SULFANILAMIDE. II. METHEMOGLOBIN FORMATION AND ITS CONTROL. PMID- 16694618 TI - BLOOD FLOW AND VASOMOTOR REACTIONS IN THE FOOT IN HEALTH, IN ARTERIOSCLEROSIS, AND IN THROMBO-ANGIITIS OBLITERANS. PMID- 16694620 TI - THE IODINE BALANCE IN NODULAR GOITER. PMID- 16694619 TI - THE INSULIN AND THE ZINC CONTENT OF NORMAL AND DIABETIC PANCREAS. PMID- 16694621 TI - THE USE OF THE SKIN TEST WITH THE TYPE SPECIFIC POLY-SACCHARIDES IN THE CONTROL OF SERUM DOSAGE IN PNEUMOCOCCAL PNEUMONIA. PMID- 16694622 TI - THE RESPONSE OF DIABETICS TO A STANDARD TEST DOSE OF INSULIN. PMID- 16694623 TI - THE DISTRIBUTION OF ASCORBIC ACID BETWEEN CELLS AND SERUM IN RELATION TO ITS URINARY EXCRETION. PMID- 16694624 TI - THE EXCRETION OF PORPHYRINS IN CONGENITAL PORPHYRIA. PMID- 16694625 TI - THE MECHANISM OF THE EXCRETION OF VITAMIN C BY THE HUMAN KIDNEY. PMID- 16694626 TI - STUDIES IN TEMPERATURE SENSATION. IV. THE STIMULATION OF COLD SENSATION BY RADIATION. PMID- 16694627 TI - THE COAGULATION DEFECT IN HEMOPHILIA. STUDIES ON THE REFRACTORY PHASE FOLLOWING REPEATED INJECTIONS OF GLOBULIN SUBSTANCE DERIVED FROM NORMAL HUMAN PLASMA IN HEMOPHILIA. PMID- 16694629 TI - CARBOHYDRATE TOLERANCE AFTER PROTAMINE INSULIN: ITS BEARING ON THE PHYSIOLOGY OF INSULIN SECRETION. PMID- 16694628 TI - A DIRECT METHOD FOR THE ESTIMATION OF SKIN DISTENSIBILITY WITH ITS APPLICATION TO THE STUDY OF VASCULAR STATES. PMID- 16694630 TI - EXCHANGE OF SUBSTANCES IN AQUEOUS SOLUTION BETWEEN JOINTS AND THE VASCULAR SYSTEM. PMID- 16694631 TI - METABOLISM IN IDIOPATHIC STEATORRHEA. I. THE INFLUENCE OF DIETARY AND OTHER FACTORS ON LIPID AND MINERAL BALANCE. PMID- 16694632 TI - METABOLISM IN IDIOPATHIC STEATORRHEA. II. EFFECT OF LIVER EXTRACT AND VITAMIN D ON CALCIUM, PHOSPHORUS, NITROGEN, AND LIPID BALANCES. PMID- 16694633 TI - RENAL FUNCTION AS A FACTOR IN THE URINARY EXCRETION OF ASCORBIC ACID. PMID- 16694634 TI - THE SIGNIFICANCE OF PROLONGED STREPTOCOCCAL ANTIBODY DEVELOPMENT IN RHEUMATIC FEVER. PMID- 16694635 TI - THE PROPHYLACTIC USE OF SULFANILAMIDE IN STREPTOCOCCAL RESPIRATORY INFECTIONS, WITH ESPECIAL REFERENCE TO RHEUMATIC FEVER. PMID- 16694636 TI - THE GASTRIC SECRETION IN CHRONIC ALCOHOLIC ADDICTION. PMID- 16694637 TI - STUDIES ON THE PHYSIOLOGY OF RESPIRATION IN PREGNANCY. EFFECTS OF BARBITURATES ADMINISTERED DURING LABOR. PMID- 16694638 TI - A COMPARISON OF THE EFFECTS OF VITAMIN D, DIHYDRO-TACHYSTEROL (A.T. 10), AND PARATHYROID EXTRACT ON THE DISORDERED METABOLISM OF RICKETS. PMID- 16694639 TI - THE EFFECT OF HIGH INTRACRANIAL VENOUS PRESSURE UPON THE CEREBRAL CIRCULATION AND ITS RELATION TO CEREBRAL SYMPTOMS. PMID- 16694640 TI - THE MECHANISM OF GLUCOSE EXCRETION BY THE KIDNEY IN DIABETIC DOGS. PMID- 16694641 TI - A DIRECT METHOD FOR THE DETERMINATION OF VENOUS PRESSURE; RELATIONSHIP OF TISSUE PRESSURE TO VENOUS PRESSURE. PMID- 16694642 TI - STUDIES IN SERUM ELECTROLYTES. XII. THE EFFECT OF WATER RESTRICTION IN A PATIENT WITH ADDISON'S DISEASE RECEIVING SODIUM CHLORIDE. PMID- 16694643 TI - A LONG TERM STUDY OF THE VARIATION OF SERUM CHOLESTEROL IN MAN. PMID- 16694644 TI - PROTEINURIA FOLLOWING MOMENTARY VASCULAR CONSTRICTION. PMID- 16694645 TI - CLINICAL STUDIES OF THE BLOOD VOLUME. V. HYPERTHYROIDISM AND MYXEDEMA. PMID- 16694646 TI - URINARY EXCRETION OF ANDROGENIC SUBSTANCES AFTER INTRAMUSCULAR AND ORAL ADMINISTRATION OF TESTOSTERONE PROPIONATE TO HUMANS. PMID- 16694647 TI - THE RATE OF FILTRATION THROUGH THE CAPILLARY WALLS AS MEASURED BY THE PRESSURE PLETHYSMOGRAPH. OBSERVATIONS ON CONTROL SUBJECTS AND ON PATIENTS WITH INTRAHEPATIC DISEASE, THYROTOXICOSIS, AND MYXEDEMA. PMID- 16694648 TI - THE LIPID DISTRIBUTION OF HUMAN PLATELETS IN HEALTH AND DISEASE. PMID- 16694649 TI - THE NATURE OF THE HUMAN FACTOR IN INFANTILE PARALYSIS. I. PECULARITIES OF GROWTH AND DEVELOPMENT. PMID- 16694650 TI - THE NATURE OF THE HUMAN FACTOR IN INFANTILE PARALYSIS. II. RELATION OF AGE TO MATURING ACHIEVEMENT AND THE DISEASE PICTURE. PMID- 16694651 TI - GONADOTROPIC HORMONE: URINE ASSAYS OF NORMALLY CYCLING, MENOPAUSAL, CASTRATED, AND ESTRIN TREATED HUMAN FEMALES. PMID- 16694652 TI - THE CONTROL OF METHEMOGLOBINEMIA WITH METHYLENE BLUE. PMID- 16694653 TI - WATER EXCHANGE OF PREMATURE INFANTS-COMPARISON OF METABOLIC (ORGANIC) AND ELECTROLYTE (INORGANIC) METHODS OF MEASUREMENT. PMID- 16694654 TI - EXPERIMENTAL HYPOSTHENURIA. PMID- 16694655 TI - FURTHER OBSERVATIONS ON THE OCCURRENCE OF RHEUMATIC MANIFESTATIONS IN THE FAMILIES OF RHEUMATIC PATIENTS. PMID- 16694656 TI - AUTOLOGOUS AND HOMOLOGOUS TRANSFUSION OF HUMAN ASCITIC FLUID. PMID- 16694657 TI - BLOOD FLOW AND VASOMOTOR REACTIONS IN THE HAND, FOREARM, FOOT, AND CALF IN RESPONSE TO PHYSICAL AND CHEMICAL STIMULI. PMID- 16694658 TI - ELECTROLYTE BALANCES DURING ARTIFICIAL FEVER WITH SPECIAL REFERENCE TO LOSS THROUGH THE SKIN. PMID- 16694659 TI - CHANGES IN PULMONARY VOLUME FOLLOWING LOBECTOMY FOR BRONCHIECTASIS. PMID- 16694660 TI - YEAST AS AN EXTRINSIC FACTOR IN RELATION TO PERNICIOUS ANEMIA. PMID- 16694661 TI - SEROLOGICAL DIFFERENTIATION OF OBSTRUCTIVE FROM HEPATOGENOUS JAUNDICE BY FLOCCULATION OF CEPHALIN-CHOLESTEROL EMULSIONS. PMID- 16694662 TI - THE EFFECT OF SLEEP ON SKIN TEMPERATURE REACTIONS IN A CASE OF ACROCYANOSIS. PMID- 16694663 TI - THE INFLUENCE OF THE FOODSTUFFS UPON THE SUSCEPTIBILITY OF THE LIVER TO INJURY BY CHLOROFORM, AND THE PROBABLE MECHANISM OF THEIR ACTION. PMID- 16694664 TI - THE NATURE OF LEUKEMIC BLOOD CELLS AS DETERMINED BY THEIR METABOLISM. PMID- 16694665 TI - THE ANTIKETOGENIC ACTIVITY OF SUCCINIC ACID. PMID- 16694666 TI - IMMUNOLOGICAL STUDIES IN PATIENTS WITH PNEUMOCOCCUS TYPE III PNEUMONIA TREATED WITH SUL-FANILAMIDE AND SERUM. PMID- 16694668 TI - THE ELIMINATION OF CHOLIC ACIDS. IV. IN PATIENTS WITH LIVER DISEASES. PMID- 16694667 TI - THE EFFECTS OF SPINAL ANESTHESIA ON THE CIRCULATION IN NORMAL, UNOPERATED MAN WITH REFERENCE TO THE AUTONOMY OF THE ARTERIOLES, AND ESPECIALLY THOSE OF THE RENAL CIRCULATION. PMID- 16694669 TI - THE ORIGIN AND NATURE OF NORMAL SYNOVIAL FLUID. PMID- 16694670 TI - A SIMPLE METHOD FOR DETERMINING THE SYSTOLIC BLOOD PRESSURE OF THE UNANESTHETIZED RAT. PMID- 16694671 TI - RESPONSES OF NORMAL SUBJECTS AND OF PATIENTS WITH DIABETES INSIPIDUS TO WATER AND SALT INGESTION. PMID- 16694672 TI - LACTIC ACID PRODUCTION DURING REST AND AFTER EXERCISE IN SUBJECTS WITH VARIOUS TYPES OF HEART DISEASE WITH SPECIAL REFERENCE TO CONGENITAL HEART DISEASE. PMID- 16694673 TI - THE USE OF CO(2) INHALATION AS A TEST OF CIRCULATION TIME. PMID- 16694674 TI - CLINICAL STUDIES OF THE BLOOD VOLUME. VI. CHANGES IN BLOOD VOLUME IN PERNICIOUS ANEMIA IN RELATION TO THE HEMATOPOIETIC RESPONSE TO INTRA-MUSCULAR LIVER EXTRACT THERAPY. PMID- 16694675 TI - A SECRETORY DEPRESSANT IN GASTRIC JUICE OF PATIENTS WITH PERNICIOUS ANEMIA. PMID- 16694676 TI - SULFAPYRIDINE, SULFANILAMIDE, AND SPECIFIC ANTISERUM IN EXPERIMENTAL TYPE III PNEUMOCOCCIC INFECTIONS. PMID- 16694677 TI - ON VAGAL AND EXTRAVAGAL FACTORS IN CARDIAC SLOWING BY DIGITALIS IN PATIENTS WITH AURICULAR FIBRILLATION. PMID- 16694678 TI - MECHANISM OF THE ARTERIAL HYPERTENSION INDUCED BY PAREDRINOL (alpha-N-DIMETHYL-p HYDROXYPHENETHYLAMINE. PMID- 16694679 TI - TREATMENT OF ADDISON'S DISEASE WITH DESOXY-CORTICOSTERONE ACETATE, A SYNTHETIC ADRENAL CORTICAL HORMONE (PRELIMINARY REPORT). PMID- 16694680 TI - THE MAGNESIUM CONTENT OF THE ERYTHROCYTES IN PERNICIOUS AND SOME OTHER ANEMIAS. PMID- 16694681 TI - THE RATE OF ATTAINMENT OF DIFFUSION EQUILIBRIUM FOR THIOCYANATE BETWEEN PLASMA AND TRANSUDATES FOLLOWING THE INTRAVENOUS INJECTION OF SODIUM THIOCYANATE IN PATIENTS WITH EDEMA. PMID- 16694682 TI - THE FORMATION OF METHEMOGLOBIN AND SULFHEMOGLOBIN DURING SULFANILAMIDE THERAPY. PMID- 16694683 TI - THE IN VITRO FORMATION OF AN OXIDIZING AGENT BY SURVIVING TISSUES AND SULFANILAMIDE. PMID- 16694684 TI - CLINICAL STUDIES OF THE BLOOD VOLUME. VII. CHANGES IN BLOOD VOLUME IN BRIGHT'S DISEASE WITH OR WITHOUT EDEMA, RENAL INSUFFICIENCY, OR CONGESTIVE HEART FAILURE, AND IN HYPERTENSION. PMID- 16694685 TI - INABILITY TO DEMONSTRATE A PLATELET REDUCING SUBSTANCE IN AN ACETONE EXTRACT OF THE SPLEEN FROM PATIENTS WITH IDIOPATHIC THROMBOCYTOPENIC PURPURA. PMID- 16694686 TI - STUDIES IN IRON TRANSPORTATION AND METABOLISM. III. THE NORMAL FLUCTUATIONS OF SERUM AND "EASILY SPLIT-OFF" BLOOD IRON IN INDIVIDUAL SUBJECTS. PMID- 16694687 TI - STUDIES IN IRON TRANSPORTATION AND METABOLISM. IV. OBSERVATIONS ON THE ABSORPTION OF IRON FROM THE GASTRO-INTESTINAL TRACT. PMID- 16694688 TI - THE TREATMENT OF ACUTE ALCOHOLISM WITH GLUCOSE AND INSULIN. PMID- 16694689 TI - THE BLOOD V-FACTOR (COENZYME) LEVEL IN NORMAL AND PATHOLOGICAL SUBJECTS. PMID- 16694690 TI - TREATMENT OF GAS GANGRENE INFECTIONS IN GUINEA-PIGS WITH NEOPRONTOSIL, SULFANILAMIDE, AND SULFAPYRIDINE: AN EXPERIMENTAL STUDY. PMID- 16694691 TI - ULTRAFILTRABLE MAGNESIUM IN HYPERTHYROIDISM. PMID- 16694692 TI - THE COAGULATION DEFECT IN HEMOPHILIA: THE CLOT PROMOTING ACTIVITY IN HEMOPHILIA OF BERKEFELDED NORMAL HUMAN PLASMA FREE FROM FIBRINOGEN AND PROTHROMBIN. PMID- 16694693 TI - THE OCCURRENCE OF ABNORMAL DARK ADAPTATION AND ITS RELATION TO VITAMIN A METABOLISM IN PATIENTS WITH CIRRHOSIS OF THE LIVER. PMID- 16694694 TI - THE VARIABILITY OF PROTEINURIA IN THE HYPERTENSIVE COMPLICATIONS OF PREGNANCY. PMID- 16694696 TI - THE NON-SPECIFICITY OF SUSPENSIONS OF SODIUM XANTHINE IN PROTECTING THE LIVER AGAINST INJURY BY CHLOROFORM, AND THE PROBABLE CAUSE OF ITS ACTION. PMID- 16694695 TI - CLINICAL STUDIES OF THE BLOOD VOLUME. VIII. MACROCYTIC AND HYPOCHROMIC ANEMIAS DUE TO CHRONIC BLOOD LOSS, HEMOLYSIS AND MISCELLANEOUS CAUSES, AND POLYCYTHEMIA VERA. PMID- 16694697 TI - THE EFFECT OF QUANTITATIVE REDUCTION OF RENAL BLOOD FLOW UPON THE PREGNANT RABBIT. PMID- 16694698 TI - PERIPHERAL VASCULAR ACTION OF ESTROGEN IN THE HUMAN MALE. PMID- 16694699 TI - STUDIES ON DESTRUCTION OF RED BLOOD CELLS. II. CHRONIC HEMOLYTIC ANEMIA WITH PAROXYSMAL NOCTURNAL HEMOGLOBINURIA: CERTAIN IMMUNOLOGICAL ASPECTS OF THE HEMOLYTIC MECHANISM WITH SPECIAL REFERENCE TO SERUM COMPLEMENT. PMID- 16694700 TI - EFFECT OF PITRESSIN IN CIRCULATORY COLLAPSE INDUCED BY SODIUM NITRITE. PMID- 16694701 TI - EFFECT OF PAREDRINOL (alpha-N-DIMETHYL-p-HYDROXYPHENETHYLAMINE) ON SODIUM NITRITE COLLAPSE AND ON CLINICAL SHOCK. PMID- 16694702 TI - BLOOD CHEMICAL CHANGES IN BOECK'S SARCOID WITH PARTICULAR REFERENCE TO PROTEIN, CALCIUM AND PHOSPHATASE VALUES. PMID- 16694703 TI - CAPILLARY BLOOD PRESSURE IN MAN. COMPARISON OF DIRECT AND INDIRECT METHODS OF MEASUREMENT. PMID- 16694704 TI - THE VITAMIN C REQUIREMENT OF MAN. ESTIMATED AFTER PROLONGED STUDIES OF THE PLASMA CONCENTRATION AND DAILY EXCRETION OF VITAMIN C IN 3 ADULTS ON CONTROLLED DIETS. PMID- 16694705 TI - EFFECT OF ESTROGENIC SUBSTANCE ON THE BLOOD SUGAR OF FEMALE DIABETICS AFTER THE MENOPAUSE. PMID- 16694706 TI - POSTMORTEM HEPATIC GLYCOGENOLYSIS IN HYPERINSULINISM AND GLYCOGEN DISEASE. PMID- 16694707 TI - THE FRACTIONATION OF THE IODINE OF THE BLOOD IN THYROID DISEASE. PMID- 16694708 TI - SERUM LIPOIDS AND PROTEINS IN HYPOTHYROIDISM. PMID- 16694709 TI - THE EFFECT OF PROLONGED ADMINISTRATION OF SULFANILAMIDE ON RATS WITH NEPHROTOXIC NEPHRITIS. PMID- 16694710 TI - CALCIUM EXCHANGE OF THE INTESTINE IN EXPERIMENTAL HYPERTHYROIDISM. PMID- 16694711 TI - DIET AND DEATH IN ACUTE UREMIA. PMID- 16694712 TI - A STUDY OF ACUTE RENAL INSUFFICIENCY. PMID- 16694713 TI - CHANGES IN THE GLUCOSE TOLERANCE OF OBESE SUBJECTS AFTER WEIGHT REDUCTION. PMID- 16694714 TI - THE RELATIONSHIP BETWEEN THE ERYTHROCYTE SEDIMENTATION RATE AND THE PLASMA PROTEINS. PMID- 16694715 TI - SPECIFIC VOLUME OF PLASMA AND SERUM PROTEINS IN PREGNANT AND IN PARTURIENT WOMEN AND THEIR NEWBORN CHILDREN AS DERIVED FROM VISCOSITY MEASUREMENTS. PMID- 16694716 TI - OBSERVATIONS ON THE ABSORPTION, DISTRIBUTION AND EXCRETION OF SULPHAPYRIDINE, DAGENAN OR M. & B. 693. PMID- 16694717 TI - THE COAGULATION DEFECT IN HEMOPHILIA: STUDIES OF THE CLOT PROMOTING ACTIVITY ASSOCIATED WITH PLASMA EUGLOBULIN IN HEMOPHILIA. PMID- 16694718 TI - SERUM PHOSPHATASE IN THE LYMPHOMATOID DISEASES. PMID- 16694719 TI - THE CONTROL OF THE DOSAGE OF ANTISERUM IN THE TREATMENT OF PNEUMOCOCCAL PNEUMONIA. II. THE CLINICAL APPLICATION OF THE FRANCIS SKIN TEST. PMID- 16694720 TI - THE OCCURRENCE OF METHEMOGLOBINEMIA DURING SULFANILAMIDE THERAPY. PMID- 16694721 TI - EFFECT OF DISEASE OF THE LIVER AND BILIARY TRACT UPON THE PHOSPHATASE ACTIVITY OF THE SERUM. PMID- 16694722 TI - METABOLIC DISTURBANCES IN EXPERIMENTAL HUMAN VITAMIN B DEFICIENCY. PMID- 16694723 TI - STUDIES ON THE ACTION OF SULFAPYRIDINE ON PNEUMOCOCCI. PMID- 16694724 TI - IMMUNOLOGICAL STUDIES ON PATIENTS WITH PNEUMOCOCCIC PNEUMONIA TREATED WITH SULFAPYRIDINE. PMID- 16694725 TI - A COMPARATIVE STUDY OF THE BLOOD CONCENTRATIONS AND URINARY EXCRETION OF SULFAPYRIDINE AND SULFANILAMIDE AFTER SINGLE DOSES OF SULFAPYRIDINE AND RELATED COMPOUNDS ADMINISTERED BY VARIOUS ROUTES. PMID- 16694726 TI - BACTERICIDAL ACTION OF SODIUM SULFAPYRIDINE AND OF A GLUCOSE-SULFAPYRIDINE SOLUTION IN HUMAN BLOOD. PMID- 16694727 TI - THE DIODRAST CLEARANCE AND RENAL BLOOD FLOW IN TOXEMIAS OF PREGNANCY. PMID- 16694728 TI - OBSERVATIONS ON THE ETIOLOGIC RELATIONSHIP OF ACHYLIA GASTRICA TO PERNICIOUS ANEMIA. VIII. FURTHER STUDIES OF THE PROTEOLYTIC ACTIVITY OF NORMAL HUMAN GASTRIC JUICE IN VITRO; AND THE LIMITATIONS OF THE METHOD IN PERNICIOUS ANEMIA. PMID- 16694729 TI - RENAL FUNCTION AND THE AZOTEMIA FOLLOWING HEMATEMESIS. PMID- 16694730 TI - IMMUNITY IN DIABETES. III. RELATION OF TISSUE GLYCOGEN AND BLOOD CHEMISTRY TO BACTERIAL DISSEMINATION, ANTIBODY FORMATION AND SURVIVAL AFTER INFECTION IN DIABETES. PMID- 16694731 TI - NATURE OF PERIPHERAL RESISTANCE IN ARTERIAL HYPERTENSION. PMID- 16694732 TI - THE URINARY EXCRETION OF RIBOFLAVIN FLUOROMETRIC METHODS FOR ITS ESTIMATION. PMID- 16694733 TI - SULFAPYRIDINE IN EXPERIMENTAL PNEUMOCOCCIC PNEUMONIA IN THE DOG. PMID- 16694734 TI - THE CIRCULATION IN ATHLETES. PMID- 16694735 TI - SERUM LIPOIDS AND PROTEINS IN HYPERTHYROIDISM. PMID- 16694736 TI - THE DECREASE OF GASTRIC SECRETION WITH ADVANCING YEARS: FURTHER OBSERVATIONS. PMID- 16694737 TI - THE STREPTOCOCCAL ANTIFIBRINOLYSIN TEST IN CLINICAL USE. PMID- 16694738 TI - THE ANTISTREPTOLYSIN TITER IN RHEUMATIC FEVER, ARTHRITIS AND OTHER DISEASES. PMID- 16694739 TI - THE RELATION OF METHEMOGLOBIN TO THE CYANOSIS OBSERVED AFTER SULFANILAMIDE ADMINISTRATION. PMID- 16694740 TI - THE RENAL EXCRETION OF INULIN AT LOW PLASMA CONCENTRATIONS OF THIS COMPOUND, AND ITS RELATIONSHIP TO THE GLOMERULAR FILTRATION RATE IN NORMAL, NEPHRITIC AND HYPERTENSIVE INDIVIDUALS. PMID- 16694741 TI - PRECIPITIN STUDIES IN NEPHROSIS AND NEPHRITIS. PMID- 16694742 TI - THE CONTROL OF THE DOSAGE OF ANTISERUM IN THE TREATMENT OF PNEUMOCOCCAL PNEUMONIA. I. A STUDY OF THE MECHANISM OF THE SKIN REACTION TO TYPE SPECIFIC POLYSACCHARIDE. PMID- 16694743 TI - STUDIES ON NEOPLASMS WITH THE AID OF RADIOACTIVE PHOSPHORUS. I. THE TOTAL PHOSPHORUS METABOLISM OF NORMAL AND LEUKEMIC MICE. PMID- 16694744 TI - A COMPARISON OF PROCEDURES FOR INCREASING BLOOD FLOW TO LIMBS USING AN IMPROVED OPTICAL PLETHYSMOGRAPH. PMID- 16694745 TI - THE DETERMINATION OF THE GLOMERULAR FILTRATION BY THE ENDOGENOUS CREATININE CLEARANCE. PMID- 16694746 TI - STUDIES ON THE CONDITIONS OF GLUCOSE EXCRETION IN MAN. PMID- 16694747 TI - ANTIBODY FORMATION IN CASES OF LOBAR PNEUMONIA TREATED WITH SULFAPYRIDINE. PMID- 16694748 TI - ELECTROPHORETIC ANALYSIS OF PLASMA AND URINARY PROTEINS. PMID- 16694749 TI - THE EFFECT OF UNILATERAL SPONTANEOUS PNEUMOTHORAX ON THE CIRCULATION IN MAN. PMID- 16694750 TI - CALCIUM AND PHOSPHORUS METABOLISM IN OSTEOMALACIA IX. METABOLIC BEHAVIOR OF INFANTS FED ON BREAST MILK FROM MOTHERS SHOWING VARIOUS STATES OF VITAMIN D NUTRITION. PMID- 16694751 TI - CALCIUM AND PHOSPHORUS METABOLISM IN OSTEOMALACIA X. FURTHER STUDIES ON VITAMIN D ACTION: EARLY SIGNS OF DEPLETION AND EFFECT OF MINIMAL DOSES. PMID- 16694752 TI - OBSERVATIONS ON THE PLASMA PROTHROMBIN AND THE EFFECTS OF VITAMIN K IN PATIENTS WITH LIVER OR BILIARY TRACT DISEASE. PMID- 16694753 TI - OBSERVATIONS ON THE SERUM CHOLESTEROL IN ACUTE INFECTIONS AS RECORDED DURING AND AFTER PNEUMONIA. PMID- 16694754 TI - FOUR RECENT INFLUENZA EPIDEMICS: AN EXPERIMENTAL STUDY. PMID- 16694755 TI - STUDIES OF RHEUMATIC DISEASE. III. FAMILIAL ASSOCIATION AND AGGREGATION IN RHEUMATIC DISEASE. PMID- 16694756 TI - URINARY EXCRETION OF THIAMIN IN CLINICAL CASES AND THE VALUE OF SUCH ANALYSES IN THE DIAGNOSIS OF THIAMIN DEFICIENCY. PMID- 16694757 TI - THE GLYCINE TOLERANCE TEST IN SPRUE AND PERNICIOUS ANEMIA. PMID- 16694758 TI - A STUDY OF SULFUR METABOLISM AND THE EFFECT OF SULFUR ADMINISTRATION IN CHRONIC ARTHRITIS. PMID- 16694759 TI - BALLISTOCARDIOGRAM. II. NORMAL STANDARDS, ABNORMALITIES COMMONLY FOUND IN DISEASES OF THE HEART AND CIRCULATION, AND THEIR SIGNIFICANCE. PMID- 16694760 TI - INCREASED UROBILINOGEN EXCRETION AND ACUTE HEMOLYTIC ANEMIA IN PATIENTS TREATED WITH SULFAPYRIDINE. PMID- 16694761 TI - DISTRIBUTION OF ASCORBIC ACID IN THE BLOOD AND ITS NUTRITIONAL SIGNIFICANCE. PMID- 16694762 TI - FACTORS THAT INFLUENCE THE PASSAGE OF ASCORBIC ACID FROM SERUM TO CELLS IN HUMAN BLOOD. PMID- 16694763 TI - RENAL BLOOD FLOW IN WOMEN WITH HYPERTENSION AND RENAL IMPAIRMENT. PMID- 16694764 TI - CORRELATION BETWEEN THE CONCENTRATION OF BISULPHITE BINDING SUBSTANCES IN THE BLOOD AND THE URINARY THIAMIN EXCRETION. PMID- 16694765 TI - THE INFLUENCE OF VITAMIN A UPON UREA CLEARANCE IN THE HUMAN SUBJECT. PMID- 16694766 TI - BLOOD CHANGES INDUCED BY VENESECTION IN WOMEN WITH TOXEMIA OF LATE PREGNANCY. PMID- 16694767 TI - THE EFFECT OF BENZEDRINE (beta-PHENYLISOPROPYLAMINE SULPHATE) AND PAREDRINE (rho HYDROXY -alpha-METHYL-PHENYLETHYLAMINE HYDROBROMIDE) ON THE CIRCULATION, METABOLISM AND RESPIRATION IN NORMAL MAN. PMID- 16694768 TI - THE EFFECT OF PAREDRINE ON THE VENOUS SYSTEM. PMID- 16694769 TI - OBSERVATIONS ON THE BLOOD IODINE. I. THE BLOOD IODINE IN HEALTH, IN THYROID AND CARDIORENAL DISEASE, AND IN LEUKEMIA. PMID- 16694770 TI - RENAL FUNCTION IN PATIENTS WITH GOUT. PMID- 16694771 TI - THE RELATION OF SPECIFIC GRAVITY TO COMPOSITION AND TOTAL SOLIDS IN NORMAL HUMAN URINE. PMID- 16694772 TI - IODINE COMPONENTS OF THE BLOOD. CIRCULATING THYROGLOBULIN IN NORMAL PERSONS AND IN PERSONS WITH THYROID DISEASE. PMID- 16694773 TI - THE EFFECT OF THE APPLICATION OF TOURNIQUETS ON THE HEMODYNAMICS OF THE CIRCULATION. PMID- 16694774 TI - ON THE EXACERBATION IN CHRONIC GLOMERULONEPHRITIS. PMID- 16694775 TI - STUDIES ON THE INTRAPULMONARY MIXTURE OF GASES. I. NITROGEN ELIMINATION FROM BLOOD AND BODY TISSUES DURING HIGH OXYGEN BREATHING. PMID- 16694776 TI - STUDIES ON THE INTRAPULMONARY MIXTURE OF GASES. II. ANALYSIS OF THE REBREATHING METHOD (CLOSED CIRCUIT) FOR MEASURING RESIDUAL AIR. PMID- 16694777 TI - STUDIES ON THE INTRAPULMONARY MIXTURE OF GASES. III. AN OPEN CIRCUIT METHOD FOR MEASURING RESIDUAL AIR. PMID- 16694778 TI - STUDIES ON ANACIDITY: THE HYDROGEN-ION CONCENTRATION OF THE GASTRIC SECRETION, THE GASTROSCOPIC APPEARANCE OF THE GASTRIC MUCOSA, AND THE PRESENCE OF A GASTRIC SECRETORY DEPRESSANT IN PATIENTS WITH ANACIDITY. PMID- 16694779 TI - THE EFFECT OF EXERCISE ON THE VOLUME OF THE BLOOD. PMID- 16694780 TI - A STUDY OF THE EFFECTS OF SULFANILAMIDE ON ACID-BASE METABOLISM. PMID- 16694781 TI - SERUM URATE IN RELATIVES OF GOUTY PATIENTS. PMID- 16694782 TI - STUDIES ON PAIN. A NEW METHOD FOR MEASURING PAIN THRESHOLD: OBSERVATIONS ON SPATIAL SUMMATION OF PAIN. PMID- 16694783 TI - STUDIES ON PAIN. MEASUREMENT OF THE EFFECT OF MORPHINE, CODEINE, AND OTHER OPIATES ON THE PAIN THRESHOLD AND AN ANALYSIS OF THEIR RELATION TO THE PAIN EXPERIENCE. PMID- 16694784 TI - THE AMYLOLYTIC AND PHOSPHATASE ACTIVITY OF LIVER TISSUE IN VON GIERKE'S DISEASE. PMID- 16694785 TI - THE MECHANISM OF THE EXCRETION OF VITAMIN C BY THE HUMAN KIDNEY AT LOW AND NORMAL PLASMA LEVELS OF ASCORBIC ACID. PMID- 16694787 TI - ON THE INACTIVATION OF THROMBIN BY PLASMA PROTEIN. PMID- 16694786 TI - A PHOTOELECTRIC METHOD FOR THE QUANTITATIVE DETERMINATION OF ERYTHROCYTE FRAGILITY. PMID- 16694788 TI - ALKALOSIS AND LOW PLASMA POTASSIUM IN A CASE OF CUSHING'S SYNDROME: A METABOLIC STUDY. PMID- 16694789 TI - EFFECTIVENESS OF PER-ORAL INSULIN IN HUMAN DIABETES. PMID- 16694790 TI - BRONCHIAL CALIBRE CHANGES IN BRONCHIECTASIS. PMID- 16694791 TI - STUDIES IN RHEUMATIC DISEASE. V. THE AGE AT ONSET OF PRIMARY RHEUMATIC ATTACK. PMID- 16694792 TI - ANTIBODY FORMATION IN CASES OF LOBAR PNEUMONIA TREATED WITH SULFATHIAZOLE. PMID- 16694793 TI - RELATIONS OF EFFECTIVE RENAL BLOOD FLOW AND GLOMERULAR FILTRATION TO TUBULAR EXCRETORY MASS IN NORMAL MAN. PMID- 16694794 TI - GLOMERULAR DYNAMICS IN THE NORMAL HUMAN KIDNEY. PMID- 16694795 TI - THE ORIGIN AND NATURE OF NORMAL HUMAN SYNOVIAL FLUID. PMID- 16694796 TI - EXCRETION OF SODIUM PREGNANEDIOL GLUCURONIDATE IN URINE OF NORMAL HUMAN PREGNANCY. PMID- 16694797 TI - THE EFFECT OF LARGE DOSES OF VITAMINS A, B, C AND D ON THE INCIDENCE OF UPPER RESPIRATORY INFECTIONS IN A GROUP OF RHEUMATIC CHILDREN. PMID- 16694798 TI - CARBOHYDRATE METABOLISM IN ADDISON'S DISEASE. PMID- 16694799 TI - CHOLINE ESTERASE OF BLOOD CELLS AND PLASMA IN BLOOD DYSCRASIAS, WITH SPECIAL REFERENCE TO PERNICIOUS ANEMIA. PMID- 16694800 TI - FAT METABOLISM IN DIABETES MELLITUS. PMID- 16694801 TI - BASAL GASTRIC SECRETION AS A CLINICAL TEST OF GASTRIC FUNCTION WITH SPECIAL REFERENCE TO PEPTIC ULCER. PMID- 16694802 TI - MECHANISM OF DIURESIS: ALTERATIONS IN THE SPECIFIC GRAVITY OF THE BLOOD PLASMA WITH ONSET OF DIURESIS IN HEART FAILURE. PMID- 16694803 TI - STUDIES ON THE RELIEF OF PAIN BY COUNTERIRRITATION. PMID- 16694804 TI - A QUANTITATIVE STUDY OF THE URINARY EXCRETION OF HYPOPHYSEAL GONADOTROPIN, ESTROGEN, AND ANDROGEN OF NORMAL WOMEN. PMID- 16694805 TI - EFFECT OF VARYING INTAKE OF PROTEIN AND SALTS ON THE COMPOSITION AND SPECIFIC GRAVITY OF URINE. PMID- 16694806 TI - DISTRIBUTION OF ASCORBIC ACID BETWEEN CELLS AND SERUM OF HUMAN BLOOD. PMID- 16694807 TI - THE RENAL EXCRETION OF INORGANIC PHOSPHATE IN RELATION TO THE ACTION OF VITAMIN D AND PARATHYROID HORMONE. PMID- 16694808 TI - STUDIES ON NEOPLASMS WITH THE AID OF RADIOACTIVE PHOSPHORUS. II. THE PHOSPHORUS METABOLISM OF THE NUCLEOPROTEIN, PHOSPHOLIPID AND ACID SOLUBLE FRACTIONS OF NORMAL AND LEUKEMIC MICE. PMID- 16694809 TI - MEASUREMENT OF THE EFFECT ON THE PAIN THRESHOLD OF ACETYLSALICYLIC ACID, ACETANILID, ACETOPHENETIDIN, AMINOPYRINE, ETHYL ALCOHOL, TRICHLORETHYLENE, A BARBITURATE, QUININE, ERGOTAMINE TARTRATE AND CAFFEINE: AN ANALYSIS OF THEIR RELATION TO THE PAIN EXPERIENCE. PMID- 16694810 TI - THE URINARY/FECAL COPROPORPHYRIN RATIO IN LIVER DISEASE. PMID- 16694811 TI - THROMBOCYTOPEN: A CONFIRMATORY REPORT. PMID- 16694812 TI - THE MEASUREMENT OF CARDIAC OUTPUT. AN IMPROVEMENT OF THE ACETYLENE METHOD PROVIDING AN INHERENT CHECK. PMID- 16694813 TI - THE LIVER LIPIDS IN NORMAL HUMAN LIVERS AND IN CASES OF CIRRHOSIS AND FATTY INFILTRATION OF THE LIVER. PMID- 16694814 TI - ELECTROPHORETIC ANALYSIS OF THE PROTEINS OF PLASMA AND SEROUS EFFUSIONS. PMID- 16694815 TI - THE RENAL BLOOD FLOW IN COARCTATION OF THE AORTA. PMID- 16694816 TI - MEASUREMENTS OF BLOOD FLOW AND BLOOD PRESSURE IN CLUBBED FINGERS. PMID- 16694817 TI - THE TOXICITY OF ORALLY ADMINISTERED POTASSIUM SALTS IN RENAL INSUFFICIENCY. PMID- 16694818 TI - THE EFFECTS OF THE PATENT DUCTUS ARTERIOSUS ON THE CIRCULATION. PMID- 16694819 TI - THE CARDIAC OUTPUT AND OTHER MEASUREMENTS OF THE CIRCULATION IN COARCTATION OF THE AORTA. PMID- 16694820 TI - LYSOLECITHIN AND HEMOLYTIC ANEMIA. THE SIGNIFICANCE OF LYSOLECITHIN PRODUCTION IN THE DIFFERENTIATION OF CIRCULATING AND STAGNANT BLOOD. PMID- 16694821 TI - THE PATHOGENESIS OF AZOTEMIA IN HEMORRHAGE FROM THE UPPER GASTRO-INTESTINAL TRACT. PMID- 16694822 TI - ON THE RENAL TUBULAR EXCRETION OF CREATININE IN NORMAL MAN. PMID- 16694823 TI - STUDIES OF HEMOGLOBINEMIA AND HEMOGLOBINURIA PRODUCED IN MAN BY INTRAVENOUS INJECTION OF HEMOGLOBIN SOLUTIONS. PMID- 16694824 TI - OBSERVATIONS ON THE INHIBITION OF SULFONAMIDE ACTION BY PARA-AMINOBENZOIC ACID. PMID- 16694825 TI - A DEFECT IN THE METABOLISM OF TYROSINE AND PHENYLALANINE IN PREMATURE INFANTS. I. IDENTIFICATION AND ASSAY OF INTERMEDIARY PRODUCTS. PMID- 16694826 TI - A DEFECT IN THE METABOLISM OF TYROSINE AND PHENYLALANINE IN PREMATURE INFANTS. II. SPONTANEOUS OCCURRENCE AND ERADICATION BY VITAMIN C. PMID- 16694827 TI - HYDROGEN ION CONCENTRATION OF THE DUODENAL CONTENTS UNDER FASTING CONDITIONS IN NORMAL PERSONS AND IN PATIENTS WITH DUODENAL ULCER: A COMPARATIVE STUDY. PMID- 16694828 TI - A STUDY OF URINARY RIBOFLAVIN EXCRETION IN MAN. PMID- 16694829 TI - STUDIES ON INFECTIOUS MONONUCLEOSIS. II. THE RELATIONSHIP OF THE ORGANISMS OF THE GENUS LISTERELLA TO THE DISEASE, AS STUDIED BY THE AGGLUTINATION REACTION. PMID- 16694830 TI - THE CEPHALIN-CHOLESTEROL FLOCCULATION TEST AS AN AID IN THE DIAGNOSIS OF HEPATIC DISORDERS. PMID- 16694831 TI - FURTHER OBSERVATIONS ON SKIN REACTIONS TO ANTIGENS FROM HETEROLOGOUS CESTODES IN ECHINOCOCCUS DISEASE. PMID- 16694832 TI - CALCIUM AND PHOSPHORUS METABOLISM IN OSTEOMALACIA. XI. THE PATHOGENETIC ROLE OF PREGNANCY AND RELATIVE IMPORTANCE OF CALCIUM AND VITAMIN D SUPPLY. PMID- 16694833 TI - OBSERVATIONS ON THE EFFECT OF STREPTOCOCCAL UPPER RESPIRATORY INFECTIONS ON RHEUMATIC CHILDREN: A THREE-YEAR STUDY. PMID- 16694834 TI - AN EPIDEMIC OF INFLUENZA B OCCURRING IN A GROUP OF RHEUMATIC CHILDREN CONCURRENT WITH AN OUTBREAK OF STREPTOCOCCAL PHARYNGITIS: CLINICAL AND EPIDEMIOLOGICAL OBSERVATIONS. PMID- 16694835 TI - STUDIES OF CALCIUM AND PHOSPHORUS METABOLISM. XVI. THE INFLUENCE OF THE PITUITARY GLAND. PMID- 16694836 TI - ACID-BASE BALANCE, RENAL FUNCTION, AND GASTRIC SECRETION DURING HYPOCHLOREMIA IN THE DOG. PMID- 16694837 TI - CLINICAL STUDIES ON PYRIDOXINE (VITAMIN B(6)). PMID- 16694838 TI - DEMONSTRATION THAT THE CELL PLASMA RATIO OF BLOOD CONTAINED IN MINUTE VESSELS IS LOWER THAN THAT OF VENOUS BLOOD. PMID- 16694839 TI - THE CLEARANCE OF BILIRUBIN FROM THE PLASMA. A MEASURE OF THE EXCRETING POWER OF THE LIVER. PMID- 16694840 TI - MEASUREMENT OF THE SENSITIVITY OF THE SMALLEST BLOOD VESSELS IN HUMAN SKIN: RESPONSES TO GRADED MECHANICAL STIMULATION IN NORMAL MEN. PMID- 16694841 TI - BLOOD IODINE IN PATIENTS WITH THYROID DISEASE. PMID- 16694842 TI - LUNG VOLUME UNDER SURGICAL ANAESTHESIA: THE EFFECT OF AVERTIN ON THE SUBTIDAL AIR. PMID- 16694843 TI - THE RELATION OF HIGH AND LOW UREA CLEARANCES TO THE INULIN AND CREATININE CLEARANCES IN CHILDREN WITH THE NEPHROTIC SYNDROME. PMID- 16694844 TI - SPONTANEOUS RECOVERY FROM NUTRITIONAL MACROCYTIC ANEMIA IN YOUNG SWINE FOLLOWING INITIAL ESTRUS. PMID- 16694845 TI - STUDIES ON THE CHEMICAL DIAGNOSIS OF PELLAGRA (NICOTINIC ACID DEFICIENCY). PMID- 16694846 TI - VITAMIN A AND CAROTENE. I. THE DETERMINATION OF VITAMIN A IN THE BLOOD AND LIVER AS AN INDEX OF VITAMIN A NUTRITION OF THE RAT. PMID- 16694847 TI - VITAMIN A AND CAROTENE. II. VITAMIN A AND CAROTENE METABOLISM IN DIABETICS AND NORMALS. PMID- 16694848 TI - METABOLISM, TOXICITY AND MANNER OF ACTION OF GOLD COMPOUNDS USED IN THE TREATMENT OF ARTHRITIS. I. HUMAN PLASMA AND SYNOVIAL FLUID CONCENTRATION AND URINARY EXCRETION OF GOLD DURING AND FOLLOWING TREATMENT WITH GOLD SODIUM THIOMALATE, GOLD SODIUM THIOSULFATE, AND COLLOIDAL GOLD SULFIDE. PMID- 16694849 TI - THE LIVER LIPIDS AND THEIR DISTRIBUTION IN DISEASE. AN ANALYSIS OF 60 HUMAN LIVERS. PMID- 16694850 TI - STUDIES ON BLOOD HISTAMINE IN PATIENTS WITH ALLERGY. II. ALTERATIONS IN THE BLOOD HISTAMINE IN PATIENTS WITH ALLERGIC DISEASE. PMID- 16694851 TI - MAGNESIUM PARTITION STUDIES IN GRAVES' DISEASE AND IN CLINICAL AND EXPERIMENTAL HYPOTHYROIDISM. PMID- 16694852 TI - INFLUENCES OF ERYTHROCYTES AND OF LEUKOCYTES ON STABILITY AND TRANSFER OF ASCORBIC ACID IN HUMAN BLOOD. PMID- 16694853 TI - EVIDENCE FOR GENERAL DISTRIBUTION OF PERIPHERAL RESISTANCE IN COARCTATION OF THE AORTA: REPORT OF THREE CASES. PMID- 16694854 TI - TREATMENT OF CIRRHOSIS OF THE LIVER BY A NUTRITIOUS DIET AND SUPPLEMENTS RICH IN VITAMIN B COMPLEX. PMID- 16694855 TI - THE INTRAVENOUS GLUCOSE TOLERANCE TEST. PMID- 16694856 TI - THE PERIPHERAL VASCULAR RESPONSE TO EXERCISE IN THE HYPERTHYROID STATE. PMID- 16694857 TI - STUDIES ON PAIN. OBSERVATIONS ON PAIN DUE TO LOCAL COOLING AND ON FACTORS INVOLVED IN THE "COLD PRESSOR" EFFECT. PMID- 16694858 TI - FORMULAE FOR AFFERENT AND EFFERENT ARTERIOLAR RESISTANCE IN THE HUMAN KIDNEY: AN APPLICATION TO THE EFFECTS OF SPINAL ANESTHESIA. PMID- 16694859 TI - THE RELATIVE CHANGES IN AFFERENT AND EFFERENT ARTERIOLAR RESISTANCE IN THE NORMAL HUMAN KIDNEY. PMID- 16694860 TI - MECHANISMS OF RESPIRATORY FAILURE UNDER BARBITURATE ANESTHESIA (EVIPAL, PENTOTHAL). PMID- 16694861 TI - CLINICAL STUDIES WITH THE AID OF RADIOACTIVE PHOSPHORUS. I. THE ABSORPTION AND DISTRIBUTION OF RADIO-PHOSPHORUS IN THE BLOOD AND ITS EXCRETION BY NORMAL INDIVIDUALS AND PATIENTS WITH LEUKEMIA. PMID- 16694862 TI - STUDIES ON NEOPLASMS WITH THE AID OF RADIOACTIVE PHOSPHORUS. III. THE PHOSPHORUS METABOLISM OF THE PHOSPHOLIPID, ACID SOLUBLE AND NUCLEOPROTEIN FRACTIONS OF VARIOUS TISSUES OF NORMAL AND LEUKEMIC MICE FOLLOWING THE ADMINISTRATION OF "TRACER" AND "THERAPEUTIC" DOSES OF RADIO-PHOSPHORUS. PMID- 16694863 TI - EXPERIMENTAL AND CLINICAL STUDIES ON GRAMICIDIN. PMID- 16694864 TI - ACUTE HEMOLYTIC ANEMIA FROM THE SULFONAMIDES. PMID- 16694865 TI - SPECTROPHOTOMETRY OF FAIRLEY'S NEW BLOOD PIGMENT, METHEMALBUMIN. PMID- 16694866 TI - THE CAUSE OF DEATH IN EXPERIMENTAL ANURIA. PMID- 16694867 TI - THE THIAMIN CONTENT OF HUMAN BLOOD AND URINE AS DETERMINED BY THE FERMENTATION METHOD. PMID- 16694868 TI - NOTE ON THE INTERPRETATION OF CLEARANCE METHODS IN THE DISEASED KIDNEY. PMID- 16694869 TI - EFFECTIVE RENAL BLOOD FLOW IN SUBJECTS WITH ESSENTIAL HYPERTENSION. PMID- 16694870 TI - EFFECTIVE RENAL BLOOD FLOW IN THE SEPARATE KIDNEYS OF SUBJECTS WITH ESSENTIAL HYPERTENSION. PMID- 16694871 TI - AN EPIDEMIC OF INFLUENZA. RESULTS OF PROPHYLACTIC INOCULATION OF A COMPLEX INFLUENZA A-DISTEMPER VACCINE. PMID- 16694872 TI - CIRCULATORY FAILURE IN ACUTE INFECTIONS. PMID- 16694873 TI - STUDIES ON INTRAPULMONARY MIXTURE OF GASES. IV. THE SIGNIFICANCE OF THE PULMONARY EMPTYING RATE AND A SIMPLIFIED OPEN CIRCUIT MEASUREMENT OF RESIDUAL AIR. PMID- 16694874 TI - A COMPARISON BETWEEN DEHYDRATION FROM SALT LOSS AND FROM WATER DEPRIVATION. PMID- 16694875 TI - CALCULATION OF THE VENOUS-ARTERIAL SHUNT IN CONGENITAL HEART DISEASE. PMID- 16694876 TI - THE PLASMA LEVELS OF VITAMIN A AFTER THE INGESTION OF STANDARD DOSES: STUDIES IN NORMAL SUBJECTS AND PATIENTS WITH CIRRHOSIS OF THE LIVER. PMID- 16694877 TI - THE HEMODYNAMIC EFFECTS OF ANGIOTONIN IN NORMAL MAN. PMID- 16694878 TI - THE NATURE OF THE ARTERIAL HYPERTENSION PRODUCED IN NORMAL SUBJECTS BY THE ADMINISTRATION OF ANGIOTONIN. PMID- 16694879 TI - SOME PROPERTIES OF HUMAN FETAL AND MATERNAL BLOOD. PMID- 16694880 TI - METABOLIC STUDIES IN PATIENTS WITH CANCER OF THE GASTRO-INTESTINAL TRACT. I. PLASMA VITAMIN A LEVELS IN PATIENTS WITH MALIGNANT NEOPLASTIC DISEASE, PARTICULARLY OF THE GASTRO-INTESTINAL TRACT. PMID- 16694881 TI - FRACTIONATION OF SERUM PROTEINS IN HYPERPROTEINEMIA, WITH SPECIAL REFERENCE TO MULTIPLE MYELOMA. PMID- 16694882 TI - THE CARDIO-CIRCULATORY EFFECTS IN MAN OF NEO-SYNEPHRIN (1-alpha-hydroxy-beta methylamino-3-hydroxy-ethylbenzene hydrochloride). PMID- 16694883 TI - THE ACID-BASE BALANCE OF PREMATURE INFANTS. PMID- 16694885 TI - THE CARDIO-CIRCULATORY EFFECTS IN MAN OF SYNEPHRIN TARTRATE (dl-alpha-hydroxy beta-methylamino-4-hydroxy-ethylbenzene hydrochloride). PMID- 16694884 TI - RENAL FUNCTION IN PATIENTS WITH ADDISON'S DISEASE AND IN PATIENTS WITH ADRENAL INSUFFICIENCY SECONDARY TO PITUITARY PAN-HYPOFUNCTION. PMID- 16694886 TI - THE CHANGES IN THE BLOOD PRESSURE AND IN THE RENAL BLOOD FLOW AND GLOMERULAR FILTRATION RATE OF HYPERTENSIVE PATIENTS FOLLOWING UNILATERAL NEPHRECTOMY. PMID- 16694887 TI - RADIOACTIVE IODINE AS AN INDICATOR IN THYROID PHYSIOLOGY. IV. THE METABOLISM OF IODINE IN GRAVES' DISEASE. PMID- 16694888 TI - RADIOACTIVE IODINE AS AN INDICATOR IN THYROID PHYSIOLOGY. V. THE USE OF RADIOACTIVE IODINE IN THE DIFFERENTIAL DIAGNOSIS OF TWO TYPES OF GRAVES' DISEASE. PMID- 16694889 TI - THE ROLE OF THE ADRENAL CORTEX IN ACUTE ANOXIA. PMID- 16694890 TI - EFFECTS OF INTERRUPTING AND RESTORING THE CIRCULATION TO THE LOWER EXTREMITIES. PMID- 16694891 TI - THE FILTRATION RATE, EFFECTIVE RENAL BLOOD FLOW, TUBULAR EXCRETORY MASS AND PHENOL RED CLEARANCE IN NORMAL PREGNANCY. PMID- 16694892 TI - THE FILTRATION RATE, EFFECTIVE RENAL BLOOD FLOW, TUBULAR EXCRETORY MASS AND PHENOL RED CLEARANCE IN SPECIFIC TOXEMIA OF PREGANANCY. PMID- 16694893 TI - THE PREVENTION OF SENSORY NEURON DEGENERATION IN THE PIG, WITH SPECIAL REFERENCE TO THE ROLE OF VARIOUS LIVER FRACTIONS. PMID- 16694894 TI - PATHOLOGICAL VARIATIONS IN BLOOD AND SPINAL FLUID PYRUVIC ACID. PMID- 16694895 TI - THE SIGNIFICANCE OF PORPHYRINURIA IN LEAD POISONING. PMID- 16694896 TI - STUDIES ON THE INFLUENCE OF VITAMIN A AND VITAMIN C ON CERTAIN IMMUNOLOGICAL REACTIONS IN MAN. PMID- 16694897 TI - THE INFLUENCE OF DIET ON THE ASCORBIC ACID REQUIREMENT OF PREMATURE INFANTS. PMID- 16694898 TI - BLEEDING TIME, LYMPH TIME, AND CLOT RESISTANCE IN MEN. PMID- 16694899 TI - THE SIGNIFICANCE OF THE D: N RATIO AND ITS BEARING ON THE MECHANISM OF DIABETES MELLITUS. PMID- 16694900 TI - THE MEASUREMENT AND METABOLISM OF THIAMIN AND OF A PYRIMIDINE STIMULATING YEAST FERMENTATION FOUND IN THE BLOOD CELLS AND URINE OF NORMAL INDIVIDUALS. PMID- 16694901 TI - THE MEASUREMENT AND METABOLISM OF THIAMIN AND OF A PYRIMIDINE STIMULATING YEAST FERMENTATION FOUND IN THE BLOOD CELLS AND URINE OF PATIENTS WITH LEUKEMIA. PMID- 16694902 TI - FURTHER STUDIES ON INCREASED SUSCEPTIBILITY TO CHLOROFORM POISONING PRODUCED IN THE ALBINO RAT BY INJECTION OF CRYSTALLINE THYROXIN. PMID- 16694903 TI - THE EFFECTS OF LARGE INTRAVENOUS INFUSIONS ON BODY FLUID. PMID- 16694904 TI - INTRAVENOUS MAGNESIUM SULFATE IN THE TREATMENT OF NEPHRITIC CONVULSIONS IN ADULTS. PMID- 16694905 TI - THE SEROLOGICAL TYPING OF HEMOLYTIC STREPTOCOCCI OF THE LANCEFIELD GROUP A. PMID- 16694906 TI - CARDIAC OUTPUT, BLOOD AND INTERSTITIAL FLUID VOLUMES, TOTAL CIRCULATING SERUM PROTEIN, AND KIDNEY FUNCTION DURING CARDIAC FAILURE AND AFTER IMPROVEMENT. PMID- 16694907 TI - THE EFFECT OF FOREIGN SURFACES ON BLOOD COAGULATION. PMID- 16694908 TI - THE URINARY EXCRETION OF THIAMINE AS AN INDEX OF THE NUTRITIONAL LEVEL: ASSESSMENT OF THE VALUE OF A TEST DOSE. PMID- 16694909 TI - OSMOMETRIC BEHAVIOR OF NORMAL HUMAN ERYTHROCYTES. PMID- 16694910 TI - THE EXCRETION OF SPECIFIC FLUORESCENT SUBSTANCES IN THE URINE IN EXPERIMENTAL NICOTINIC ACID DEFICIENCY. PMID- 16694911 TI - A STUDY OF THE REFLEX MECHANISM OF SWEATING IN THE HUMAN BEING; EFFECT OF ANESTHESIA AND SYMPATHECTOMY. PMID- 16694912 TI - THE METABOLISM OF AMINO ACIDS IN DIABETES MELLITUS. PMID- 16694913 TI - CHEMOTHERAPY OF PNEUMOCOCCIC MENINGITIS WITH SPECIAL REFERENCE TO SULFATHIAZOLE. PMID- 16694914 TI - COMPARISON OF RESULTS OF THE NORMAL BALLISTOCARDIOGRAM AND A DIRECT FICK METHOD IN MEASURING THE CARDIAC OUTPUT IN MAN. PMID- 16694915 TI - THE IN VITRO AND IN VIVO EFFECT OF SULFONAMIDES UPON THE STREPTOCOCCAL ANTIFIBRINOLYSIN TEST. PMID- 16694916 TI - LATENT LIVER DISEASE IN PERSONS RECOVERED FROM CATARRHAL JAUNDICE AND IN OTHERWISE NORMAL MEDICAL STUDENTS AS REVEALED BY THE BILIRUBIN EXCRETION TEST. PMID- 16694917 TI - VITAMIN A DEFICIENCY IN LAENNEC'S CIRRHOSIS. THE RELATIVE SIGNIFICANCE OF THE PLASMA VITAMIN A AND CAROTENOID LEVELS AND THE DARK ADAPTATION TIME. PMID- 16694918 TI - THE EFFECT OF EXTERNAL PRESSURE ON THE VASCULAR VOLUME OF THE FOREARM AND ITS RELATION TO CAPILLARY BLOOD PRESSURE AND VENOUS PRESSURE. PMID- 16694919 TI - THE EFFECT OF EPINEPHRINE ON THE VOLUME OF THE BLOOD. PMID- 16694920 TI - COMPLEMENT ACTIVITY IN PNEUMONIA. PMID- 16694921 TI - THE COAGULASE TEST FOR STAPHYLOCOCCI AND ITS CORRELATION WITH THE RESISTANCE OF THE ORGANISMS TO THE BACTERICIDAL ACTION OF HUMAN BLOOD. PMID- 16694922 TI - CALCULATION OF HEAT PRODUCTION FROM INSENSIBLE LOSS OF WEIGHT. PMID- 16694923 TI - THE EFFECTS ON THE CARDIOVASCULAR SYSTEM OF FLUIDS ADMINISTERED INTRAVENOUSLY IN MAN. IV. THE LUNG VOLUME AND PULMONARY DYNAMICS. PMID- 16694924 TI - REDUCTION OF BLOOD PRESSURE ASSOCIATED WITH THE PYROGENIC REACTION IN HYPERTENSIVE SUBJECTS. PMID- 16694925 TI - THE RELATION BETWEEN DARK ADAPTATION AND THE LEVEL OF VITAMIN A IN THE BLOOD. PMID- 16694926 TI - CIRRHOSIS OF THE LIVER AMONG RATS RECEIVING DIETS POOR IN PROTEIN AND RICH IN FAT. PMID- 16694927 TI - ON THE OCCURRENCE OF DYSPNEA, DIZZINESS AND PRECORDIAL DISTRESS OCCASIONED BY THE POOLING OF BLOOD IN VARICOSE VEINS. PMID- 16694928 TI - TISSUE THIAMIN CONCENTRATIONS AND URINARY THIAMIN EXCRETION. PMID- 16694929 TI - THE EFFECT OF EXERCISE AND OF FOUR COMMONLY USED DRUGS ON THE NORMAL HUMAN ELECTROCARDIOGRAM, WITH PARTICULAR REFERENCE TO T WAVE CHANGES. PMID- 16694930 TI - ABSENCE OF BENEFICIAL EFFECTS FROM INJECTIONS OF DESOXYCORTICOSTERONE ACETATE AND OF CORTICAL ADRENAL EXTRACT IN EXPERIMENTAL ANURIA. PMID- 16694931 TI - PROTECTIVE ACTIVITY OF NORMAL HUMAN AND ANIMAL SERA FOR SULFAPYRIDINE-TREATED MICE INFECTED WITH PNEUMOCOCCI. PMID- 16694932 TI - EFFECTS OF BARBITURATE ANESTHESIA (EVIPAL AND PENTOTHAL SODIUM) UPON THE INTEGRATION OF RESPIRATORY CONTROL MECHANISMS. A STUDY DIRECTED TOWARD IMPROVEMENT OF METHODS FOR THE PRECLINICAL EVALUATION OF ANESTHETIC AGENTS. PMID- 16694933 TI - THE ANTICOAGULANT EFFECTS IN RABBITS AND MAN OF THE INTRAVENOUS INJECTION OF SALTS OF THE RARE EARTHS. PMID- 16694934 TI - A COMPARISON OF PITUITRIN WITH THE ANTIDIURETIC SUBSTANCE FOUND IN HUMAN URINE AND PLACENTA. PMID- 16694935 TI - STUDIES WITH RADIOACTIVE DI-AZO DYES. 1. THE LOCALIZATION OF RADIOACTIVE DI-BROM TRYPAN BLUE IN INFLAMMATORY LESIONS. PMID- 16694936 TI - THE SERUM ANTISTREPTOLYSIN TITER IN CHRONIC GLOMERULONEPHRITIS. PMID- 16694937 TI - THE RELATION OF THE SERUM ANTISTREPTOLYSIN TITER TO THE EXACERBATION IN CHRONIC GLOMERULONEPHRITIS. PMID- 16694938 TI - THE UREA CLEARANCE OF YOUNG PREMATURE AND FULL TERM INFANTS. PMID- 16694939 TI - INTUBATION STUDIES OF THE HUMAN SMALL INTESTINE. XXII. AN IMPROVED TECHNIC FOR THE STUDY OF ABSORPTION; ITS APPLICATION TO ASCORBIC ACID. PMID- 16694940 TI - SPECIFIC ANTIPNEUMOCOCCAL IMMUNITY IN RELATION TO THE CHEMOTHERAPY OF PNEUMONIA. PMID- 16694941 TI - STUDIES ON BLOOD COAGULATION: A PROTEOLYTIC ENZYME PREPARED FROM CALCIUM AND PLATELET FREE NORMAL HUMAN BLOOD PLASMA. PMID- 16694942 TI - STUDIES ON BLOOD COAGULATION: THE NATURE AND PROPERTIES OF A PROTEOLYTIC ENZYME DERIVED FROM PLASMA. PMID- 16694943 TI - THE DIGITAL BLOOD FLOW, ARTERIAL PRESSURE, AND VASCULAR RESISTANCE IN ARTERIAL HYPERTENSION AND IN CORONARY THROMBOSIS. PMID- 16694944 TI - THE DIGITAL CIRCULATION IN PERIPHERAL VASCULAR DISEASES. PMID- 16694945 TI - THE MEASUREMENT AND RECORDING OF GASTRODUODENAL BLOOD FLOW IN MAN BY MEANS OF A THERMAL GRADIENTOMETER. PMID- 16694946 TI - ADRENAL CORTICAL HYPERPLASIA WITH VIRILISM: DIAGNOSIS, COURSE AND TREATMENT. PMID- 16694947 TI - AN ELECTROPHORETIC STUDY OF THE PROTEIN COMPONENTS IN CEREBROSPINAL FLUID AND THEIR RELATIONSHIP TO THE SERUM PROTEINS. PMID- 16694948 TI - OBSERVATIONS ON THE NATURE OF MYASTHENIA GRAVIS. THE EFFECT OF THYMECTOMY ON NEURO-MUSCULAR TRANSMISSION. PMID- 16694949 TI - EFFECTS PRODUCED BY THE INTRAVENOUS INJECTION IN MAN OF A TOXIC ANTIGENIC MATERIAL DERIVED FROM EBERTHELLA TYPHOSA: CLINICAL, HEMATOLOGICAL, CHEMICAL AND SEROLOGICAL STUDIES. PMID- 16694950 TI - THE PRODUCTION OF CARDIAC LESIONS BY REPEATED INJECTIONS OF DESOXYCORTICOSTERONE ACETATE. PMID- 16694951 TI - THE VELOCITY OF BLOOD FLOW IN INFANTS AND YOUNG CHILDREN, DETERMINED BY RADIOACTIVE SODIUM. PMID- 16694952 TI - A STUDY OF THE INFLUENCE OF THE CHARACTER OF AN EXAMINING ROOM ON THE PERIPHERAL BLOOD VESSELS OF NORMAL, HYPERTENSIVE, AND SENILE SUBJECTS. PMID- 16694953 TI - REACTIVITY OF INTACT BLOOD VESSELS OF THE FINGERS AND TOES TO SENSORY STIMULI IN NORMAL RESTING ADULTS, IN PATIENTS WITH HYPERTENSION, AND IN SENILE SUBJECTS. PMID- 16694954 TI - BLOOD FLOW IN THE HAND AND FOREARM AFTER PARAVERTEBRAL BLOCK OF THE SYMPATHETIC GANGLIA. EVIDENCE AGAINST SYMPATHETIC VASODILATOR NERVES IN THE EXTREMITIES OF MAN. PMID- 16694955 TI - SENSITIVITY OF THE SMALLEST CUTANEOUS BLOOD VESSELS: QUANTITATIVE RESPONSES TO GRADED MECHANICAL STIMULATION AND TO LOCAL ISCHEMIA IN ARTERIAL HYPERTENSION, ARTERIOSCLEROSIS, AND CERTAIN ALLIED DISORDERS. PMID- 16694956 TI - THE EFFECTS ON RENAL RESISTANCE TO BLOOD FLOW OF RENIN, ANGIOTONIN, PITRESSIN AND ATROPINE, HYPERTENSION, AND TOXEMIA OF PREGNANCY. PMID- 16694957 TI - CAPILLARY BLOOD PRESSURE IN MAN. DIRECT MEASUREMENTS IN THE DIGITS DURING INDUCED VASOCONSTRICTION. PMID- 16694958 TI - CAPILLARY BLOOD PRESSURE IN MAN. DIRECT MEASUREMENTS IN THE DIGITS OF NORMAL AND HYPERTENSIVE SUBJECTS DURING VASOCONSTRICTION AND VASODILATATION VARIOUSLY INDUCED. PMID- 16694959 TI - CAPILLARY BLOOD PRESSURE IN MAN. DIRECT MEASUREMENTS IN THE DIGITS DURING ARTERIAL HYPERTENSION INDUCED BY PAREDRINOL SULFATE. PMID- 16694960 TI - THE INFLUENCE OF ALTERATIONS IN ACID-BASE BALANCE UPON TRANSFERS OF CARBON DIOXIDE AND BICARBONATE IN MAN. PMID- 16694961 TI - EFFECT OF MUSCULAR EXERCISE UPON THE PERIPHERAL CIRCULATION IN PATIENTS WITH VALVULAR HEART DISEASE. PMID- 16694962 TI - THE QUALITATIVE EXAMINATION OF URINARY CALCULI. PMID- 16694963 TI - THE CLASSIFICATION AND CHEMICAL PATHOGENESIS OF URINARY CALCULI. PMID- 16694964 TI - THE RELATION OF HIPPURIC ACID EXCRETION TO THE VOLUME OF THE URINE. PMID- 16694965 TI - SERUM IODINE FRACTIONS IN HYPERTHYROIDISM. PMID- 16694966 TI - SERUM MAGNESIUM IN THYROID DISEASE. PMID- 16694967 TI - PROLONGED WATER DEPRIVATION IN THE DOG. PMID- 16694968 TI - INTUBATION STUDIES OF THE HUMAN SMALL INTESTINE. XXIII. A METHOD OF DETERMINING DIGESTIVE ACTIVITY IN ANY PORTION OF THE GASTRO-INTESTINAL TRACT, WITH SOME MEASUREMENTS OF PROTEIN DIGESTION IN THE STOMACH AND SMALL INTESTINE. PMID- 16694969 TI - UREA CLEARANCE AND DIURESIS IN MAN. PMID- 16694970 TI - GASTRIC SECRETION. II. ABSORPTION OF RADIOACTIVE SODIUM FROM POUCHES OF THE BODY AND ANTRUM OF THE STOMACH OF THE DOG. PMID- 16694971 TI - STUDIES ON RENIN: THE DURATION OF THE PRESSOR EFFECT OF LARGE DOSES IN CONSCIOUS NORMAL AND RENALLY ABNORMAL DOGS. OBSERVATIONS ON ANESTHETIZED AND UREMIC DOGS, AND THE ANAPHYLACTIC AND PATHOLOGICAL EFFECTS OF PIG RENIN. PMID- 16694972 TI - GASTRIC SECRETION. III. THE ABSORPTION OF HEAVY WATER FROM POUCHES OF THE BODY AND ANTRUM OF THE STOMACH OF THE DOG. PMID- 16694973 TI - A COMMUNITY STUDY OF CARRIERS IN EPIDEMIC POLIOMYELITIS. PMID- 16694974 TI - THE COAGULATION DEFECT IN HEMOPHILIA: A COMPARISON OF THE PROTEOLYTIC ACTIVITY OF CHLOROFORM PREPARATIONS OF HEMOPHILIC AND NORMAL HUMAN PLASMA. PMID- 16694975 TI - THE RELATION OF POSTURAL HEMODILUTION TO PAROXYSMAL DYSPNEA. PMID- 16694976 TI - DISTRIBUTION OF SULFONAMIDE COMPOUNDS BETWEEN CELLS AND SERUM OF HUMAN BLOOD. PMID- 16694977 TI - OBSERVATIONS ON CERTAIN MANIFESTATIONS OF CIRCULATORY CONGESTION PRODUCED IN DOGS BY RAPID INFUSION. PMID- 16694978 TI - THE EFFECT OF CALCIUM CARBONATE, ALUMINUM PHOSPHATE, AND ALUMINUM HYDROXIDE ON MINERAL EXCRETION IN MAN. PMID- 16694979 TI - THE EFFECTIVE STIMULUS FOR INCREASED PULMONARY VENTILATION DURING MUSCULAR EXERTION. PMID- 16694980 TI - ATYPICAL PNEUMONIA AND PSITTACOSIS. PMID- 16694981 TI - BENCE-JONES PROTEINEMIA IN MULTIPLE MYELOMA. PMID- 16694982 TI - THE PREVENTION OF STREPTOCOCCAL UPPER RESPIRATORY INFECTIONS AND RHEUMATIC RECURRENCES IN RHEUMATIC CHILDREN BY THE PROPHYLACTIC USE OF SULFANILAMIDE. PMID- 16694983 TI - PHENYLPYRUVIC OLIGOPHRENIA. REPORT OF A CASE IN AN INFANT WITH QUANTITATIVE CHEMICAL STUDIES OF THE URINE. PMID- 16694984 TI - STUDIES ON EXPERIMENTAL AND CLINICAL HYPOCHLOREMIA IN MAN. PMID- 16694985 TI - STUDIES ON THE BACTERICIDAL PROPERTIES OF THE SYNOVIAL FLUID. PMID- 16694986 TI - A VISCOUS PROTEIN OBTAINED IN LARGE AMOUNT FROM THE SERUM OF A PATIENT WITH MULTIPLE MYELOMA. PMID- 16694987 TI - TUBULAR REABSORPTION OF PHOSPHATE IN THE DOG. PMID- 16694988 TI - URINARY EXCRETION AND SERUM CONCENTRATION OF INORGANIC PHOSPHATE IN MAN. PMID- 16694989 TI - STUDIES WITH RADIOACTIVE DI-AZO DYES. II. THE SYNTHESIS AND PROPERTIES OF RADIOACTIVE DI-BROM TRYPAN BLUE AND RADIOACTIVE DI-BROM EVANS BLUE. PMID- 16694990 TI - STUDIES WITH RADIOACTIVE DI-AZO DYES. III. THE DISTRIBUTON OF RADIOACTIVE DYES IN TUMOR-BEARING MICE. PMID- 16694991 TI - AMINO ACID STUDIES. II. PLASMA AMINO ACID RETENTION, AS EVIDENCE OF IMPAIRED LIVER FUNCTION. INVESTIGATIONS IN CHILDREN WITH NEPHROSIS AND LIVER DISEASE. PMID- 16694992 TI - PLASMA CLOT TENSILE STRENGTH MEASUREMENT: ITS RELATION TO PLASMA FIBRINOGEN. PMID- 16694993 TI - THE ELECTROPHORETIC ANALYSES OF THE SERUM PROTEINS IN DISEASES OF THE LIVER. PMID- 16694994 TI - URIC ACID CLEARANCE IN NORMAL PREGNANCY AND PRE-ECLAMPSIA. PMID- 16694995 TI - PNEUMOCOCCIC PNEUMONIA: THE PROGNOSTIC SIGNIFICANCE OF THE NUMBER OF PNEUMOCOCCI IN THE SPUTUM IN RELATION TO THERAPY, BACTEREMIA, TYPE, LEUKOCYTE COUNT, DURATION OF THE DISEASE, AGE, AND DEGREE OF INVOLVEMENT. PMID- 16694996 TI - TYPE III PNEUMONIA: THE PROGNOSTIC SIGNIFICANCE OF RETICULATION IN RELATION TO THE NUMBER OF PNEUMOCOCCI IN THE SPUTUM, THERAPY, BACTEREMIA, LEUKOCYTE COUNT, AGE, AND DEGREE OF INVOLVEMENT. PMID- 16694997 TI - AN ACOUSTIC RESPIROGRAPH. A METHOD FOR THE STUDY OF RESPIRATION THROUGH THE GRAPHIC RECORDING OF THE BREATH SOUNDS. PMID- 16694998 TI - INTUBATION STUDIES OF THE HUMAN SMALL INTESTINE. XXI. A METHOD FOR MEASURING INTRA-LUMINAL PRESSURES AND ITS APPLICATION TO THE DIGESTIVE TRACT. PMID- 16694999 TI - QUANTITATIVE RELATIONSHIPS BETWEEN BLOOD AND URINE KETONE LEVELS IN DIABETIC KETOSIS. PMID- 16695000 TI - METABOLIC STUDIES IN PATIENTS WITH GASTRO-INTESTINAL CANCER. IV. FAT METABOLISM, A METHOD OF STUDY. PMID- 16695001 TI - SULFONAMIDE-FAST PNEUMOCOCCI. A CLINICAL REPORT OF TWO CASES OF PNEUMONIA TOGETHER WITH EXPERIMENTAL STUDIES ON THE EFFECTIVENESS OF PENICILLIN AND TYROTHRICIN AGAINST SULFONAMIDE-RESISTANT STRAINS. PMID- 16695002 TI - NITROGEN STORAGE FOLLOWING INTRAVENOUS AND ORAL ADMINISTRATION OF CASEIN HYDROLYSATE TO INFANTS WITH ACUTE GASTRO-INTESTINAL DISTURBANCE. PMID- 16695003 TI - TRAUMATIC SHOCK. I. THE PRODUCTION OF RADIOACTIVE PLASMA PROTEIN FROM AMINO ACIDS CONTAINING RADIOACTIVE SULFUR. PMID- 16695004 TI - TRAUMATIC SHOCK. II. THE PREPARATION OF CYSTINE, METHIONINE, AND HOMOCYSTINE CONTAINING RADIOACTIVE SULFUR. PMID- 16695005 TI - TRAUMATIC SHOCK. III. A MODIFIED ELECTROSCOPE ESPECIALLY SUITED FOR MEASURING SUBSTANCES WITH LOW ENERGY RADIATION. PMID- 16695006 TI - TRAUMATIC SHOCK: IV. A STUDY OF THE PROBLEM OF THE "LOST PLASMA" IN HEMORRHAGIC SHOCK BY THE USE OF RADIOACTIVE PLASMA PROTEIN. PMID- 16695007 TI - TRAUMATIC SHOCK. V. A STUDY OF THE EFFECT OF OXYGEN ON HEMORRHAGIC SHOCK. PMID- 16695008 TI - PRODUCTION OF TETANUS TOXIN ON PEPTONE-FREE MEDIA. PMID- 16695009 TI - CONVERSION OF HYDROLYSATE TETANUS TOXIN TO TOXOID. PMID- 16695010 TI - A COMPARISON OF ANTIGENICITIES OF HYDROLYSATE AND PEPTONE TETANUS TOXOIDS IN THE GUINEA PIG. PMID- 16695011 TI - ANTITOXIN RESPONSE IN MAN TO TETANUS TOXOIDS. PMID- 16695012 TI - BLOOD AMYLASE ACTIVITY IN DISEASE OF CARBOHYDRATE METABOLISM AND IN NON-DIABETIC PANCREATIC DISEASE. PMID- 16695013 TI - NITROGEN RETENTION, CREATINURIA, AND OTHER EFFECTS OF THE TREATMENT OF SIMMONDS' DISEASE WITH METHYL TESTOSTERONE. PMID- 16695014 TI - THE MOTILITY OF THE SMALL INTESTINE IN SPRUE. PMID- 16695015 TI - INVESTIGATIONS OF MENINGOCOCCAL INFECTION. I. BACTERIOLOGICAL ASPECTS. PMID- 16695016 TI - INVESTIGATIONS OF MENINGOCOCCAL INFECTION. II. IMMUNOLOGICAL ASPECTS. PMID- 16695017 TI - INVESTIGATIONS OF MENINGOCOCCAL INFECTION. III. THE BACTERICIDAL ACTION OF NORMAL AND IMMUNE SERA FOR THE MENINGOCOCCUS. PMID- 16695018 TI - RENAL BIOPSY STUDIES CORRELATED WITH RENAL CLEARANCE OBSERVATIONS IN HYPERTENSIVE PATIENTS TREATED BY RADICAL SYMPATHECTOMY. PMID- 16695019 TI - A QUANTITATIVE STUDY OF THE URINARY EXCRETION OF HYPOPHYSEAL GONADOTROPIN, ESTROGEN, AND NEUTRAL 17-KETOSTEROIDS OF NORMAL MEN. PMID- 16695020 TI - CARBONIC ANHYDRASE IN NEWBORN INFANTS. PMID- 16695021 TI - GROUP A HEMOLYTIC STREPTOCOCCUS ANTIBODIES. I. GRIFFITH TYPE AGGLUTININ AND ANTISTREPTOLYSIN TITERS IN NORMAL MEN AND IN ACUTE INFECTIONS. PMID- 16695022 TI - GROUP A HEMOLYTIC STREPTOCOCCUS ANTIBODIES. II. GRIFFITH TYPE AGGLUTININ AND ANTISTREPTOLYSIN TITERS IN CARRIERS AND NON-CARRIERS. PMID- 16695023 TI - THE ABSORPTION, EXCRETION, AND DISTRIBUTION OF PENICILLIN. PMID- 16695024 TI - THE CHANGES WHICH ALTER RENAL OSMOTIC WORK. PMID- 16695025 TI - DIODRAST AND INULIN CLEARANCES IN NEPHROTIC CHILDREN WITH SUPERNORMAL UREA CLEARANCES. PMID- 16695026 TI - OBSERVATIONS ON THE PHYSIOLOGY AND BIOCHEMISTRY OF QUANTITATIVE BURNS. PMID- 16695027 TI - IMPROVEMENTS IN CALCULATION OF RENAL RESISTANCE TO BLOOD FLOW. CHARTS FOR OSMOTIC PRESSURE AND VISCOSITY OF BLOOD. PMID- 16695028 TI - THE RELATIONSHIP OF DEHYDRATION AND OVERHYDRATION OF THE BLOOD PLASMA TO COLLAPSE IN THE MANAGEMENT OF ARTIFICIAL FEVER THERAPY. PMID- 16695029 TI - FRACTIONATION OF THE SERUM AND PLASMA PROTEINS BY SALT PRECIPITATION IN INFANTS AND CHILDREN. 1. THE CHANGES WITH MATURITY AND AGE. 2. THE CHANGES IN GLOMERULONEPHRITIS. 3. THE CHANGES IN NEPHROSIS. PMID- 16695030 TI - THE PROLONGATION OF THE ACTION OF SUBCUTANEOUSLY INJECTED MEDICINES IN MAN. PMID- 16695031 TI - THE EFFECT OF OPIATES ON THE PAIN THRESHOLD IN POST-ADDICTS. PMID- 16695032 TI - SKIN RESISTANCE CHANGES AND MEASUREMENTS OF PAIN THRESHOLD. PMID- 16695033 TI - A STUDY OF THE VOLUME OF THE BLOOD IN CONGESTIVE HEART FAILURE. RELATION TO OTHER MEASUREMENTS IN FIFTEEN PATIENTS. PMID- 16695034 TI - QUANTITATIVE RELATIONSHIP BETWEEN BASAL METABOLIC RATE AND THYROID DOSAGE IN PATIENTS WITH TRUE MYXEDEMA. PMID- 16695035 TI - TOLERANCE TO ORAL THYROID AND REACTION TO INTRAVENOUS THYROXINE IN SUBJECTS WITHOUT MYXEDEMA. PMID- 16695036 TI - ELECTROPHORETIC AND CHEMICAL ANALYSIS OF PROTEIN IN NEPHRITIC URINE. PMID- 16695037 TI - A DEFECT IN THE METABOLISM OF TYROSINE AND PHENYLALANINE IN PREMATURE INFANTS. III. DEMONSTRATION OF THE IRREVERSIBLE CONVERSION OF PHENYLALANINE TO TYROSINE IN THE HUMAN ORGANISM. PMID- 16695038 TI - THE RELATION OF CEPHALIN FLOCCULATION AND COLLOIDAL GOLD REACTIONS TO THE SERUM PROTEINS. PMID- 16695039 TI - STUDIES REGARDING A GLUTAMINE-LIKE SUBSTANCE IN BLOOD AND SPINAL FLUID, INCLUDING A METHOD FOR ITS QUANTITATIVE DETERMINATION. PMID- 16695040 TI - EFFECT OF INSULIN HYPOGLYCEMIA AND GLUCOSE ADMINISTRATION ON THE LEVEL OF THE GLUTAMINE-LIKE SUBSTANCE IN BLOOD SERUM. PMID- 16695041 TI - THE EFFECT OF TESTOSTERONE AND ALLIED COMPOUNDS ON THE MINERAL, NITROGEN, AND CARBOHYDRATE METABOLISM OF A GIRL WITH ADDISON'S DISEASE. PMID- 16695042 TI - THE MODE OF EXCRETION OF CREATINE AND CREATINE METABOLISM IN THYROID DISEASE. PMID- 16695043 TI - THE LOCUS OF ACTION OF THE PARATHYROID HORMONE: EXPERIMENTAL STUDIES WITH PARATHYROID EXTRACT ON NORMAL AND NEPHRECTOMIZED RATS. PMID- 16695044 TI - THE TREATMENT OF BURNS BY THE CLOSED-PLASTER METHOD, WITH CERTAIN PHYSIOLOGICAL CONSIDERATIONS IMPLICIT IN THE SUCCESS OF THIS TECHNIQUE. PMID- 16695045 TI - CHANGES IN THE ELECTROPHORETIC PATTERN IN LYMPH AND SERUM IN EXPERIMENTAL BURNS. PMID- 16695046 TI - THE ROLE OF THE EXTRACELLULAR FLUID IN THE MAINTENANCE OF A NORMAL PLASMA VOLUME. PMID- 16695047 TI - THE CONCENTRATION OF VITAMIN A IN THE BLOOD PLASMA DURING PREGNANCY. PMID- 16695048 TI - PENICILLIN: ITS ANTIBACTERIAL EFFECT IN WHOLE BLOOD AND SERUM FOR THE HEMOLYTIC STREPTOCOCCUS AND STAPHYLOCOCCUS AUREUS. PMID- 16695049 TI - OBSERVATIONS ON THE URINARY EXCRETION OF SULFADIAZINE. PMID- 16695050 TI - THE ESTIMATION AND CONTROL OF POST-OPERATIVE DEHYDRATION, WITH THE AID OF HEMOGLOBIN AND PLASMA PROTEIN DETERMINATIONS. PMID- 16695051 TI - SYSTEMIC AND RENAL CIRCULATORY CHANGES FOLLOWING THE ADMINISTRATION OF ADRENIN, EPHEDRINE, AND PAREDRINOL TO NORMAL MAN. PMID- 16695052 TI - CHLORIDE METABOLISM AND PLASMA AMINO ACID LEVELS IN PRIMARY ATYPICAL PNEUMONIA. PMID- 16695053 TI - THE EFFECT OF ATROPINE UPON THE ABSORPTION OF VITAMIN A. PMID- 16695054 TI - THE INTERRELATIONS OF SERUM LIPIDS IN NORMAL PERSONS. PMID- 16695055 TI - THE INTERRELATIONS OF SERUM LIPIDS IN PATIENTS WITH THYROID DISEASE. PMID- 16695056 TI - THE INTERRELATIONS OF SERUM LIPIDS IN PATIENTS WITH DISEASES OF THE KIDNEYS. PMID- 16695057 TI - ARTERIOLAR LESIONS IN HYPERTENSION: A STUDY OF 350 CONSECUTIVE CASES TREATED SURGICALLY. AN ESTIMATION OF THE PROGNOSTIC VALUE OF MUSCLE BIOPSY. PMID- 16695058 TI - STUDIES IN CANCER. VII. ENZYME DEFICIENCY IN HUMAN AND EXPERIMENTAL CANCER. PMID- 16695059 TI - THE BINDING OF SULFONAMIDE DRUGS BY PLASMA PROTEINS. A FACTOR IN DETERMINING THE DISTRIBUTION OF DRUGS IN THE BODY. PMID- 16695060 TI - STUDIES ON THE PLASMA PROTEINS. V. THE EFFECT OF CONCENTRATED SOLUTIONS OF HUMAN AND BOVINE SERUM ALBUMIN ON BLOOD VOLUME AFTER ACUTE BLOOD LOSS IN MAN. PMID- 16695061 TI - ON THE VARIATIONS OF THE PLASMA VITAMIN A LEVEL AFTER THE ADMINISTRATION OF LARGE DOSES OF VITAMIN A IN LIVER DISEASE. PMID- 16695062 TI - CALCIUM AND PHOSPHORUS METABOLISM IN RHEUMATOID ARTHRITIS AND DEGENERATIVE JOINT DISEASE. PMID- 16695063 TI - A NEW MODIFIED PROTAMINE ZINC INSULIN: COMPARISON WITH HISTONE ZINC INSULIN, CLEAR, AND STANDARD PROTAMINE ZINC INSULINS. PMID- 16695064 TI - THE EXCRETION OF GONADOTROPHIC HORMONE BY PREPUBERAL AND ADOLESCENT GIRLS. PMID- 16695065 TI - THE LUNG VOLUME AND ITS SUBDIVISIONS IN THE UPRIGHT AND RECUMBENT POSITIONS IN PATIENTS WITH CONGESTIVE FAILURE. PULMONARY FACTORS IN THE GENESIS OF ORTHOPNEA. PMID- 16695066 TI - CLINICAL STUDIES ON INCOORDINATION OF THE CIRCULATION, AS DETERMINED BY THE RESPONSE TO ARISING. PMID- 16695067 TI - PANCREATIC FUNCTION AND DISEASE IN EARLY LIFE. I. PANCREATIC ENZYME ACTIVITY AND THE CELIAC SYNDROME. PMID- 16695068 TI - EVIDENCE THAT BODY IRRITATIONS OR EMOTIONS RETARD GASTRIC EVACUATION, NOT BY PRODUCING PYLOROSPASM BUT BY DEPRESSING GASTRIC MOTILITY. PMID- 16695069 TI - TRAUMATIC SHOCK. VI. THE EFFECT OF HEMORRHAGIC SHOCK ON THE CONCENTRATION OF RENIN AND HYPERTENSINOGEN IN THE PLASMA IN UNANESTHETIZED DOGS. PMID- 16695070 TI - EXPERIMENTAL STUDIES ON HEADACHE: PHARMACODYNAMICS OF URINE EXCRETED DURING MIGRAINE HEADACHE AND ITS RELATION TO 17-KETOSTEROID CONTENT. PMID- 16695071 TI - PHYSIOLOGICAL INTRAVASCULAR HEMOLYSIS OF EXERCISE. HEMOGLOBINEMIA AND HEMOGLOBINURIA FOLLOWING CROSS-COUNTRY RUNS. PMID- 16695072 TI - THE ANALGETIC EFFECTS OF LOW CONCENTRATIONS OF NITROUS OXIDE COMPARED IN MAN WITH MORPHINE SULPHATE. PMID- 16695073 TI - THE RELATION OF GASTRIC FUNCTION TO NAUSEA IN MAN. PMID- 16695074 TI - A STUDY OF FACTORS (EMOTIONAL) RESPONSIBLE FOR CHANGES IN THE PATTERN OF SPONTANEOUS RHYTHMIC FLUCTUATIONS IN THE VOLUME OF THE VASCULAR BED OF THE FINGER TIP. PMID- 16695075 TI - EXPERIMENTAL HYPERTONICITY: ALTERATIONS IN THE DISTRIBUTION OF BODY WATER, AND THE CAUSE OF DEATH. PMID- 16695076 TI - SOME EFFECTS OF LOW ATMOSPHERIC PRESSURE ON RATS. PMID- 16695077 TI - PLASMA VOLUME OF DOGS IN DEHYDRATION, WITH AND WITHOUT SALT LOSS. PMID- 16695079 TI - A DIRECT CURRENT DIELECTROGRAPH FOR RECORDING THE MOVEMENTS OF THE HEART. PMID- 16695078 TI - GLOMERULONEPHRITIS. A SURVEY OF THE FUNCTIONAL ORGANIZATION OF THE KIDNEY IN VARIOUS STAGES OF DIFFUSE GLOMERULONEPHRITIS. PMID- 16695080 TI - THE CHLORIDE EXCRETION TEST (CUTLER-WILDER-POWER TEST) IN ASTHENIA. PMID- 16695081 TI - REGIONAL VARIATIONS IN WATER LOSS FROM THE SKIN OF DISEASED SUBJECTS LIVING IN A SUBTROPICAL CLIMATE. PMID- 16695082 TI - THE EFFECT OF INSULIN ON THE GLUCOSE TOLERANCE OF NORMAL MAN. PMID- 16695083 TI - THE UTILIZATION OF INTRAVENOUSLY INJECTED SALT IN NORMALS AND IN PATIENTS WITH CUSHING'S SYNDROME BEFORE AND AFTER ADMINISTRATION OF DESOXYCORTICOSTERONE ACETATE. PMID- 16695084 TI - STUDIES ON INTRAPULMONARY MIXTURE OF GASES. V. FORMS OF INADEQUATE VENTILATION IN NORMAL AND EMPHYSEMATOUS LUNGS, ANALYZED BY MEANS OF BREATHING PURE OXYGEN. PMID- 16695085 TI - STUDIES OF PROTOPORPHYRIN: IV. A COMPARISON OF THE ERYTHROCYTE PROTOPORPHYRIN CONCENTRATION WITH THE RETICULOCYTE PERCENTAGE UNDER EXPERIMENTAL AND CLINICAL CONDITIONS. PMID- 16695086 TI - VARIATIONS IN CUTANEOUS AND VISCERAL PAIN SENSITIVITY IN NORMAL SUBJECTS. PMID- 16695087 TI - TRANSFERS OF INTRACELLULAR POTASSIUM IN EXPERIMENTAL DEHYDRATION. PMID- 16695088 TI - OBSERVATIONS ON THE EPIDEMIOLOGY OF STREPTOCOCCAL PHARYNGITIS AND THE RELATION OF STREPTOCOCCAL CARRIERS TO THE OCCURRENCE OF OUTBREAKS. PMID- 16695089 TI - THE OCCURRENCE OF BACTERIOSTATIC PROPERTIES IN THE BLOOD OF PATIENTS AFTER RECOVERY FROM STREPTOCOCCAL PHARYNGITIS. PMID- 16695090 TI - ON THE PATHOGENESIS OF RENAL FAILURE ASSOCIATED WITH MULTIPLE MYELOMA. ELECTROPHORETIC AND CHEMICAL ANALYSIS OF PROTEIN IN URINE AND BLOOD SERUM. PMID- 16695091 TI - ATYPICAL PNEUMONIA CAUSED BY PSITTACOSIS-LIKE VIRUSES. PMID- 16695092 TI - THE DETERIORATION OF COMPLEMENT ACTIVITY IN NORMAL HUMAN SERUM. PMID- 16695093 TI - INFLUENCE OF ANESTHESIA (ETHER, CYCLOPROPANE, SODIUM EVIPAL) ON THE CIRCULATION UNDER NORMAL AND SHOCK CONDITIONS. PMID- 16695094 TI - COMPARATIVE VALUE OF BROMSULPHALEIN, SERUM PHOSPHATASE, PROTHROMBIN TIME, AND INTRAVENOUS GALACTOSE TOLERANCE TESTS IN DETECTING HEPATIC DAMAGE PRODUCED BY CARBON TETRACHLORIDE. PMID- 16695095 TI - FACTORS IN THE RESISTANCE OF GONORRHEA TO SULFONAMIDES. PMID- 16695096 TI - IMMUNOLOGIC STUDIES IN INSULIN RESISTANCE I. REPORT OF A CASE EXHIBITING VARIATIONS IN RESISTANCE AND ALLERGY TO INSULIN. PMID- 16695097 TI - IMMUNOLOGIC STUDIES IN INSULIN RESISTANCE II. THE PRESENCE OF A NEUTRALIZING FACTOR IN THE BLOOD EXHIBITING SOME CHARACTERISTICS OF AN ANTIBODY. PMID- 16695098 TI - A STUDY OF CAPILLARY PERMEABILITY IN EXPERIMENTAL BURNS AND BURN SHOCK USING RADIOACTIVE DYES IN BLOOD AND LYMPH. PMID- 16695099 TI - TISSUE THIAMINE IN HEMORRHAGIC SHOCK. PMID- 16695100 TI - EFFECT OF SODIUM IODIDE, MAGNESIUM SULFATE, THYROXIN, AND THYROTROPIC HORMONE ON THE BLOOD MAGNESIUM PARTITION. PMID- 16695101 TI - SULFONAMIDE-RESISTANT STAPHYLOCOCCI: CORRELATION OF IN VITRO SULFONAMIDE RESISTANCE WITH SULFONAMIDE THERAPY. PMID- 16695102 TI - THE EFFECT OF THE INJECTION OF HISTAMINE INTO THE BRACHIAL ARTERY ON THE PERMEABILITY OF THE CAPILLARIES OF THE FOREARM AND HAND. PMID- 16695103 TI - THE PROTEIN CONTENT OF THE EXTRACELLULAR FLUID IN NORMAL SUBJECTS AFTER VENOUS CONGESTION AND IN PATIENTS WITH CARDIAC FAILURE, ANOXEMIA, AND FEVER. PMID- 16695104 TI - THE LEVEL OF VITAMIN A AND CAROTENE IN THE PLASMA OF RHEUMATIC SUBJECTS. PMID- 16695105 TI - SUBCLINICAL VITAMIN DEFICIENCY. IV. PLASMA THIAMIN. PMID- 16695106 TI - CHARACTERISTICS OF THE NORMAL ELECTROENCEPHALOGRAM. I. A STUDY OF THE OCCIPITAL CORTICAL POTENTIALS IN 500 NORMAL ADULTS. PMID- 16695107 TI - CHARACTERISTICS OF THE NORMAL ELECTROENCEPHALOGRAM. II. THE EFFECT OF VARYING BLOOD SUGAR LEVELS ON THE OCCIPITAL CORTICAL POTENTIALS IN ADULTS DURING QUIET BREATHING. PMID- 16695108 TI - CHARACTERISTICS OF THE NORMAL ELECTROENCEPHALOGRAM. III. THE EFFECT OF VARYING BLOOD SUGAR LEVELS ON THE OCCIPITAL CORTICAL POTENTIALS IN ADULTS DURING HYPERVENTILATION. PMID- 16695109 TI - THE EFFECTS OF PLEURAL EFFUSION ON RESPIRATION AND CIRCULATION IN MAN. PMID- 16695110 TI - THE INFLUENCE OF THE COLLAPSIBILITY OF VEINS ON VENOUS PRESSURE, INCLUDING A NEW PROCEDURE FOR MEASURING TISSUE PRESSURE. PMID- 16695111 TI - PRESERVATION OF NORMAL HUMAN PLASMA IN THE LIQUID STATE. I. A STATISTICAL STUDY OF 1751 ADMINISTRATIONS. PMID- 16695112 TI - PRESERVATION OF NORMAL HUMAN PLASMA IN THE LIQUID STATE. II. COMPARATIVE IN VITRO STUDIES ON THE PHYSIOLOGIC ACTIVITY OF LABILE CONSTITUENTS OF LIQUID AND FROZEN PLASMA. PMID- 16695113 TI - PRESERVATION OF NORMAL HUMAN PLASMA IN THE LIQUID STATE. III. STUDIES ON CHEMICAL AND PHYSICO-CHEMICAL CHANGES DURING THE SECOND YEAR OF STORAGE. PMID- 16695114 TI - PRESERVATION OF NORMAL HUMAN PLASMA IN THE LIQUID STATE. IV. STUDIES ON ISOHEMAGGLUTININ TITERS. PMID- 16695115 TI - THE EFFECT OF A SINGLE INJECTION OF CONCENTRATED HUMAN SERUM ALBUMIN ON CIRCULATING PROTEINS AND PROTEINURIA IN NEPHROSIS. PMID- 16695116 TI - SKIN TEMPERATURES OF THE EXTREMITIES OF PERSONS WITH INDUCED DEFICIENCIES OF THIAMINE, RIBOFLAVIN, AND OTHER COMPONENTS OF THE B COMPLEX. PMID- 16695117 TI - THE RENAL CIRCULATION IN SHOCK. PMID- 16695118 TI - THE FATE AND EFFECTS OF TRANSFUSED SERUM OR PLASMA IN NORMAL DOGS. PMID- 16695119 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. I. THE CHARACTERIZATION OF THE PROTEIN FRACTIONS OF HUMAN PLASMA. PMID- 16695120 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. II. ELECTROPHORETIC AND ULTRACENTRIFUGAL STUDIES OF SOLUTIONS OF HUMAN SERUM ALBUMIN AND IMMUNE SERUM GLOBULINS. PMID- 16695121 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. III. AMINO ACID COMPOSITION OF PLASMA PROTEINS. PMID- 16695122 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. IV. A STUDY OF THE THERMAL STABILITY OF HUMAN SERUM ALBUMIN. PMID- 16695123 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. V. THE INFLUENCE OF NON-POLAR ANIONS ON THE THERMAL STABILITY OF SERUM ALBUMIN. PMID- 16695124 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. VI. THE OSMOTIC PRESSURE OF PLASMA AND OF SERUM ALBUMIN. PMID- 16695125 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. VII. CONCENTRATED HUMAN SERUM ALBUMIN. PMID- 16695126 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. VIII. CLINICAL USE OF CONCENTRATED HUMAN SERUM ALBUMIN IN SHOCK, AND COMPARISON WITH WHOLE BLOOD AND WITH RAPID SALINE INFUSION. PMID- 16695127 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. IX. THE TREATMENT OF SHOCK WITH CONCENTRATED HUMAN SERUM ALBUMIN: A PRELIMINARY REPORT. PMID- 16695128 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. X. THE CONCENTRATIONS OF CERTAIN ANTIBODIES IN GLOBULIN FRACTIONS DERIVED FROM HUMAN BLOOD PLASMA. PMID- 16695129 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XI. THE USE OF CONCENTRATED NORMAL HUMAN SERUM GAMMA GLOBULIN (HUMAN IMMUNE SERUM GLOBULIN) IN THE PROPHYLAXIS AND TREATMENT OF MEASLES. PMID- 16695130 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XII. THE USE OF CONCENTRATED NORMAL HUMAN SERUM GAMMA GLOBULIN (HUMAN IMMUNE SERUM GLOBULIN) IN THE PREVENTION AND ATTENUATION OF MEASLES. PMID- 16695131 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XIII. THE SEPARATION AND CONCENTRATION OF ISOHEMAGGLUTININS FROM GROUP-SPECIFIC HUMAN PLASMA. PMID- 16695132 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XIV. APPRAISAL OF ISOHEMAGGLUTININ ACTIVITY. PMID- 16695133 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XV. THE PROTEINS CONCERNED IN THE BLOOD COAGULATION MECHANISM. PMID- 16695134 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XVI. FIBRIN CLOTS, FIBRIN FILMS, AND FIBRINOGEN PLASTICS. PMID- 16695135 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XVII. A NOTE ON THE ABSORPTION RATES OF FIBRIN FILMS IN TISSUE. PMID- 16695136 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XVIII. FIBRINOGEN COAGULUM AS AN AID IN THE OPERATIVE REMOVAL OF RENAL CALCULI. PMID- 16695137 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XIX. A NOTE ON THE USE OF FIBRINOGEN AND THROMBIN IN THE SURFACE TREATMENT OF BURNS. PMID- 16695138 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XX. THE DEVELOPMENT OF FIBRIN FOAM AS A HEMOSTATIC AGENT AND FOR USE IN CONJUNCTION WITH HUMAN THROMBIN. PMID- 16695139 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XXI. THE USE OF FIBRIN FOAM AS A HEMOSTATIC AGENT IN NEUROSURGERY: CLINICAL AND PATHOLOGICAL STUDIES. PMID- 16695140 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XXII. FIBRIN FILMS IN NEUROSURGERY, WITH SPECIAL REFERENCE TO THEIR USE IN THE REPAIR OF DURAL DEFECTS AND IN THE PREVENTION OF MENINGOCEREBRAL ADHESIONS. PMID- 16695141 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XXIII. THE EFFECTS OF FEEDING POSSIBLE BLOOD SUBSTITUTES ON SERUM PROTEIN REGENERATION AND WEIGHT RECOVERY IN THE HYPOPROTEINEMIC RAT. PMID- 16695142 TI - THE IMMUNE RESPONSE OF HUMAN BEINGS TO BRIEF INFECTIONS WITH PNEUMOCOCCUS. PMID- 16695143 TI - THIOURACIL. ITS ABSORPTION, DISTRIBUTION, AND EXCRETION. PMID- 16695144 TI - SIMULTANEOUS MEASUREMENTS OF THE BLOOD VOLUME IN MAN AND DOG BY MEANS OF EVANS BLUE DYE, T1824, AND BY MEANS OF CARBON MONOXIDE. I. NORMAL SUBJECTS. PMID- 16695145 TI - SIMULTANEOUS MEASUREMENTS OF THE BLOOD VOLUME IN MAN AND DOG BY MEANS OF EVANS BLUE DYE, T1824, AND BY MEANS OF CARBON MONOXIDE. II. UNDER ABNORMAL CONDITIONS, INCLUDING SECONDARY SHOCK. PMID- 16695146 TI - EFFECT OF BLOOD SERUM FROM PATIENTS WITH MYASTHENIA GRAVIS ON THE SYNTHESIS OF ACETYLCHOLINE IN VITRO. PMID- 16695147 TI - CHANGES IN PLASMA VOLUME AND CARDIAC OUTPUT FOLLOWING THE INTRAVENOUS INJECTION OF GELATIN, SERUM, AND PHYSIOLOGICAL SALINE SOLUTION. PMID- 16695148 TI - THE INDIRECT MEASUREMENT OF MEAN VENOUS OXYGEN TENSION DURING ANOXIA. PMID- 16695149 TI - A THERAPEUTIC AND PHARMACOLOGICAL STUDY OF SULFADIAZINE, MONOMETHYLSULFADIAZINE, AND DIMETHYLSULFADIAZINE IN LOBAR PNEUMONIA. PMID- 16695150 TI - DEPRESSANT EFFECTS OF HIGH CONCENTRATIONS OF INSPIRED OXYGEN ON ERYTHROCYTOGENESIS. OBSERVATIONS ON PATIENTS WITH SICKLE CELL ANEMIA WITH A DESCRIPTION OF THE OBSERVED TOXIC MANIFESTATIONS OF OXYGEN. PMID- 16695151 TI - HEBERDEN'S NODES: THE MECHANISM OF INHERITANCE IN HYPERTROPHIC ARTHRITIS OF THE FINGERS. PMID- 16695152 TI - THE NUTRITIVE VALUE OF HUMAN PLASMA FOR THE RAT. PMID- 16695153 TI - THE ELECTROPHORETIC PATTERNS OF NORMAL PLASMA. PMID- 16695154 TI - F(1) AND F(2) OF NAJJAR AND HOLT IN THE URINE OF NORMAL YOUNG MEN. PMID- 16695155 TI - TRAUMATIC SHOCK. VII. A STUDY OF THE PROBLEM OF THE "LOST PLASMA" IN HEMORRHAGIC, TOURNIQUET, AND BURN SHOCK BY THE USE OF RADIOACTIVE IODO-PLASMA PROTEIN. PMID- 16695156 TI - TRAUMATIC SHOCK. VIII. STUDIES IN THE THERAPY AND HEMODYNAMICS OF TOURNIQUET SHOCK. PMID- 16695157 TI - THE EFFECT OF PREGNANCY ON RENAL FUNCTION IN WOMEN WITH PRE-EXISTING ESSENTIAL HYPERTENSION AND WITH CHRONIC DIFFUSE GLOMERULONEPHRITIS. PMID- 16695158 TI - CHANGES IN RENAL FUNCTION ACCOMPANYING THE HYPERTROPHY OF THE REMAINING KIDNEY AFTER UNILATERAL NEPHRECTOMY. PMID- 16695159 TI - ABSORPTION OF FERROUS AND FERRIC RADIOACTIVE IRON BY HUMAN SUBJECTS AND BY DOGS. PMID- 16695160 TI - THE RENAL EXCRETION OF CHLORIDE AND WATER IN DIABETES INSIPIDUS. PMID- 16695161 TI - THE EFFECT OF DIODRAST ON THE NORMAL URIC ACID CLEARANCE. PMID- 16695162 TI - A METHOD OF DISTRIBUTING BETA-RADIATION TO THE RETICULO-ENDOTHELIAL SYSTEM AND ADJACENT TISSUES. PMID- 16695163 TI - THE ABSORPTION AND EXCRETION OF PENICILLIN FOLLOWING CONTINUOUS INTRAVENOUS AND SUBCUTANEOUS ADMINISTRATION. PMID- 16695164 TI - A PRINCIPLE FROM LIVER EFFECTIVE AGAINST SHOCK DUE TO BURNS. PMID- 16695165 TI - ELECTROLYTE AND FLUID STUDIES DURING WATER DEPRIVATION AND STARVATION IN HUMAN SUBJECTS, AND THE EFFECT OF INGESTION OF FISH, OF CARBOHYDRATE, AND OF SALT SOLUTIONS. PMID- 16695166 TI - THE DELETERIOUS EFFECT IN DOGS OF A DRY PROTEIN RATION. PMID- 16695167 TI - BLOOD KETONES AND SERUM LIPIDS IN STARVATION AND WATER DEPRIVATION. PMID- 16695168 TI - HEPATITIS DUE TO THE INJECTION OF HOMOLOGOUS BLOOD PRODUCTS IN HUMAN VOLUNTEERS. PMID- 16695169 TI - OBSERVATIONS ON THE ABSORPTION, APPARENT VOLUME OF DISTRIBUTION, AND EXCRETION OF THIOUREA. PMID- 16695170 TI - CHEMICAL STUDIES IN HYPERTENSION. REDUCING AND NITROGENOUS FRACTIONS IN PROTEIN FREE BLOOD FILTRATES. PMID- 16695171 TI - VARIATION IN BEHAVIOR OF BUFFY COAT CULTURES AMONG INDIVIDUALS OF DIFFERENT CONSTITUTION TYPES. PMID- 16695172 TI - SERUM ALBUMIN REGENERATION AS EFFECTED BY INTRAVENOUSLY AND ORALLY ADMINISTERED PROTEIN HYDROLYSATES. PMID- 16695174 TI - AUTOHEMAGGLUTININS -"COLD AGGLUTININS". PMID- 16695173 TI - THE EFFECT OF TESTOSTERONE COMPOUNDS UPON THE NITROGEN BALANCE AND CREATINE EXCRETION IN PATIENTS WITH THYROTOXICOSIS. PMID- 16695175 TI - STUDIES OF ACCOMMODATION OF NERVE IN PARATHYROID DEFICIENCY. PMID- 16695176 TI - ON THE DETERMINATION OF ARTERIAL OXYGEN SATURATIONS FROM SAMPLES OF "CAPILLARY" BLOOD. PMID- 16695177 TI - A NEW METHOD FOR STUDYING BREATHING, WITH OBSERVATIONS UPON NORMAL AND ABNORMAL SUBJECTS. PMID- 16695178 TI - RENAL EXCRETION OF SULFAMERAZINE. PMID- 16695179 TI - THE NATURE OF THE MECHANISM OF THE SHOCK PRODUCED BY THE INJECTION OF TRYPSIN AND THROMBIN. PMID- 16695180 TI - MEASUREMENT OF CARDIAC OUTPUT IN MAN USING THE TECHNIQUE OF CATHETERIZATION OF THE RIGHT AURICLE OR VENTRICLE. PMID- 16695181 TI - PLASMA VOLUME AND "EXTRAVASCULAR THIOCYANATE SPACE" IN PNEUMOCOCCUS PNEUMONIA. PMID- 16695182 TI - PROBABLE ERRORS IN THE SIMULTANEOUS MEASUREMENT OF SEPARATE KIDNEY FUNCTIONS. PMID- 16695183 TI - THE IMPORTANCE OF RH INHIBITOR SUBSTANCE IN ANTI-RH SERUMS. PMID- 16695184 TI - STUDIES ON THE CREATINURIA DUE TO METHYLATED STEROIDS. PMID- 16695185 TI - COLD HEMAGGLUTINATION AND COLD HEMOLYSIS. THE HEMOLYSIS PRODUCED BY SHAKING COLD AGGLUTINATED ERYTHROCYTES. PMID- 16695186 TI - A PLASMA FIBRINOGEN-INCREASING FACTOR OBTAINED FROM STERILE ABSCESSES IN DOGS. PMID- 16695187 TI - THE ELECTROPHORETIC ANALYSIS OF MATERNAL AND FETAL PLASMAS AND SERA. PMID- 16695188 TI - TRAUMATIC SHOCK. IX. PRESSOR THERAPY: THE EFFECT OF PAREDRINE (p-Hydroxy-a Methylphenylethylamine Hydrobromide) ON THE CIRCULATION IN HEMORRHAGIC SHOCK IN DOGS. PMID- 16695189 TI - THE RENAL HUMORAL PRESSOR MECHANISM IN MAN. I. PREPARATION AND ASSAY OF HUMAN RENIN, HUMAN HYPERTENSINOGEN, AND HYPERTENSIN. PMID- 16695190 TI - THE RENAL HUMORAL PRESSOR MECHANISM IN MAN. II. THE EFFECT OF TRANSITORY COMPLETE CONSTRICTION OF THE HUMAN RENAL ARTERY ON BLOOD PRESSURE AND ON THE CONCENTRATION OF RENIN, HYPERTENSINOGEN, AND HYPERTENSINASE OF RENAL ARTERIAL AND VENOUS BLOOD, WITH ANIMAL OBSERVATIONS. PMID- 16695191 TI - THE RENAL HUMORAL PRESSOR MECHANISM IN MAN. III. THE HYPERTENSINASE CONTENT OF PLASMA OF CONTROL SUBJECTS AND OF PATIENTS WITH HYPERTENSION AND OTHER DISEASES. PMID- 16695192 TI - THE RENAL HUMORAL PRESSOR MECHANISM IN MAN. IV. THE HYPERTENSINOGEN CONTENT OF THE PLASMA OF NORMAL PATIENTS AND PATIENTS WITH VARIOUS DISEASES. PMID- 16695193 TI - PLASMA PROTEIN AND HEMOGLOBIN IN THE PROTEIN-DEFICIENT RAT. A THREE-DIMENSIONAL STUDY. PMID- 16695194 TI - ABSENCE OF CLINICAL DISEASE IN SPITE OF A HIGH INCIDENCE OF CARRIERS OF GROUP A HEMOLYTIC STREPTOCOCCI OF A SINGLE TYPE; FAILURE OF TYROTHRICIN TO INFLUENCE THE CARRIER RATE. PMID- 16695195 TI - EFFECT OF BILIARY FISTULA ON BROMSULPHALEIN RETENTION, SERUM PHOSPHATASE, AND BILE PHOSPHATASE. PMID- 16695196 TI - EFFECT OF ENVIRONMENTAL TEMPERATURE ON EXPERIMENTAL TRAUMATIC SHOCK IN DOGS. PMID- 16695197 TI - THE DISTRIBUTION OF BODY FLUIDS OF DOGS IN TRAUMATIC SHOCK. PMID- 16695198 TI - THE EFFECT OF SULFADIAZINE, ANTITOXINS, GLOBULINS, AND DOG PLASMA ON DOGS IN TRAUMATIC SHOCK UNDER SODIUM PENTOBARBITAL ANESTHESIA. PMID- 16695199 TI - EFFECT OF ENVIRONMENTAL TEMPERATURE AND ANESTHESIA ON THE SURVIVAL OF TOURNIQUET SHOCK IN RABBITS. PMID- 16695200 TI - THIAMINE-DEFICIENT DIET IN TOURNIQUET SHOCK IN RATS. PMID- 16695201 TI - STUDY OF THE BODY TEMPERATURE AND WATER CONTENT IN SHOCK PRODUCED BY THE CONTINUOUS INTRAVENOUS INJECTION OF ADRENALIN, WITH AND WITHOUT ANESTHESIA. PMID- 16695202 TI - A STUDY OF THE AGGLUTININ RESPONSE IN PATIENTS WITH MENINGOCOCCIC MENINGITIS. PMID- 16695203 TI - BACTERIOSTATIC AND LYTIC ACTIONS OF PENICILLIN ON SENSITIVE AND RESISTANT STAPHYLOCOCCI. PMID- 16695204 TI - PROPERTIES OF A PENICILLIN INACTIVATOR EXTRACTED FROM PENICILLIN-RESISTANT STAPHYLOCOCCI. PMID- 16695205 TI - AN EXPERIMENTAL ATTEMPT TO TRANSMIT PRIMARY ATYPICAL PNEUMONIA IN HUMAN VOLUNTEERS. PMID- 16695206 TI - THE EFFECTS OF TESTOSTERONE AND OF TESTOSTERONE PROPIONATE ON RENAL FUNCTIONS IN MAN. PMID- 16695207 TI - INFLUENZA VIRUS ISOLATIONS AND SEROLOGICAL STUDIES MADE IN BOSTON DURING THE WINTER OF 1943-1944. PMID- 16695208 TI - STUDIES ON PRIMARY ATYPICAL PNEUMONIA. I. CLINICAL FEATURES AND RESULTS OF LABORATORY INVESTIGATIONS. PMID- 16695209 TI - STUDIES ON PRIMARY ATYPICAL PNEUMONIA. II. OBSERVATIONS CONCERNING THE RELATIONSHIP OF A NON-HEMOLYTIC STREPTOCOCCUS TO THE DISEASE. PMID- 16695210 TI - A CLINICAL REPORT ON CASES OF PRIMARY ATYPICAL PNEUMONIA CAUSED BY A NEW VIRUS. PMID- 16695211 TI - THE TREATMENT OF SUBACUTE BACTERIAL ENDOCARDITIS WITH PENICILLIN. PMID- 16695212 TI - ANEMIA, HYPOPROTEINEMIA, AND CATARACTS IN SWINE FED CASEIN HYDROLYSATE OR ZEIN. COMPARISON WITH PYRIDOXINE-DEFICIENCY ANEMIA. PMID- 16695213 TI - PROGRESSIVE ADDISONIAN PERNICIOUS ANEMIA, SUCCESSFULLY TREATED WITH INTRAVENOUS CHOLINE CHLORIDE. PMID- 16695214 TI - THE CONVERSION OF HEMATIN TO BILIRUBIN FOLLOWING INTRAVENOUS ADMINISTRATION IN HUMAN SUBJECTS. PMID- 16695215 TI - MECHANISM OF THE POSITIVE CEPHALIN-CHOLESTEROL FLOCCULATION REACTION IN HEPATITIS. PMID- 16695216 TI - SIGNIFICANCE OF CEPHALIN-CHOLESTEROL FLOCCULATION TEST IN MALARIAL FEVER. PMID- 16695217 TI - COMPARATIVE STUDIES OF THE CHEMICAL CHANGES OCCURRING IN SULFONAMIDE DRUGS DURING THERAPY IN MAN. PMID- 16695218 TI - FACTORS INFLUENCING ASCITES IN PATIENTS WITH CIRRHOSIS OF THE LIVER. PMID- 16695219 TI - THE CARDIAC OUTPUT IN MALE SUBJECTS AS MEASURED BY THE TECHNIQUE OF RIGHT ATRIAL CATHETERIZATION. NORMAL VALUES WITH OBSERVATIONS ON THE EFFECT OF ANXIETY AND TILTING. PMID- 16695220 TI - THE CARDIAC OUTPUT IN PATIENTS WITH CHRONIC ANEMIA AS MEASURED BY THE TECHNIQUE OF RIGHT ATRIAL CATHETERIZATION. PMID- 16695221 TI - THE EFFECT OF VENESECTION AND THE POOLING OF BLOOD IN THE EXTREMITIES ON THE ATRIAL PRESSURE AND CARDIAC OUTPUT IN NORMAL SUBJECTS WITH OBSERVATIONS ON ACUTE CIRCULATORY COLLAPSE IN THREE INSTANCES. PMID- 16695222 TI - THE IMPORTANCE OF COMPENSATING VASOCONSTRICTION IN UNANESTHETIZED AREAS IN THE MAINTENANCE OF BLOOD PRESSURE DURING SPINAL ANESTHESIA. PMID- 16695223 TI - THE RELATIONSHIP BETWEEN CONCENTRATION OF SULFAMERAZINE IN BODY FLUIDS AND THE RESPONSE IN TREATMENT OF MENINGOCOCCIC MENINGITIS. PMID- 16695224 TI - COMPLEMENT FIXATION IN HUMAN MALARIA USING AN ANTIGEN PREPARED FROM THE CHICKEN PARASITE PLASMODIUM GALLINACEUM. PMID- 16695225 TI - STUDIES ON THE QUANTITATIVE EVALUATION OF CERTAIN TREATMENTS IN THE HEALING OF EXPERIMENTAL THIRD DEGREE BURNS. PMID- 16695226 TI - THE EFFECT OF AMINO ACIDS ON SERUM AND URINE CREATINE. PMID- 16695227 TI - COMPARISON OF THE DIURETIC ACTION OF SODIUM DEHYDROCHOLATE AND MERCUPURIN IN MAN. PMID- 16695228 TI - THE RENAL CLEARANCES OF SUBSTITUTED HIPPURIC ACID DERIVATIVES AND OTHER AROMATIC ACIDS IN DOG AND MAN. PMID- 16695229 TI - GELATIN AS A PLASMA SUBSTITUTE: THE EFFECTS OF INTRAVENOUS INFUSION OF GELATIN ON CARDIAC OUTPUT AND OTHER ASPECTS OF THE CIRCULATION OF NORMAL PERSONS, OF CHRONICALLY ILL PATIENTS, AND OF NORMAL VOLUNTEERS SUBJECTED TO LARGE HEMORRHAGE. PMID- 16695230 TI - EFFECT OF METHYL TESTOSTERONE ON URINARY 17-KETOSTEROIDS OF ADRENAL ORIGIN. PMID- 16695231 TI - TRAUMATIC SHOCK. X. THE TREATMENT OF HEMORRHAGIC SHOCK IRREVERSIBLE TO REPLACEMENT OF BLOOD VOLUME DEFICIENCY. PMID- 16695232 TI - TRAUMATIC SHOCK. XI. INTESTINAL ABSORPTION IN HEMORRHAGIC SHOCK. PMID- 16695233 TI - COLD AGGLUTININS. I. OCCURRENCE OF COLD ISOHEMAGGLUTININS IN VARIOUS CONDITIONS. PMID- 16695234 TI - COLD AGGLUTININS. II. COLD ISOHEMAGGLUTININS IN PRIMARY ATYPICAL PNEUMONIA OF UNKNOWN ETIOLOGY WITH A NOTE ON THE OCCURRENCE OF HEMOLYTIC ANEMIA IN THESE CASES. PMID- 16695235 TI - COLD AGGLUTININS. III. OBSERVATIONS ON CERTAIN SEROLOGICAL AND PHYSICAL FEATURES OF COLD AGGLUTININS IN CASES OF PRIMARY ATYPICAL PNEUMONIA AND OF HEMOLYTIC ANEMIA. PMID- 16695236 TI - COLD AGGLUTININS. IV. CRITICAL ANALYSIS OF CERTAIN ASPECTS OF THE METHOD FOR DETERMINING COLD ISOHEMAGGLUTININS. PMID- 16695237 TI - COLD AGGLUTININS. V. DETERIORATION OF COLD ISOHEMAGGLUTININS ON STORAGE. PMID- 16695238 TI - COLD AGGLUTININS. VI. AGGLUTININS FOR AN INDIFFERENT STREPTOCOCCUS IN PRIMARY ATYPICAL PNEUMONIA AND IN OTHER CONDITIONS AND THEIR RELATION TO COLD ISOHEMAGGLUTININS. PMID- 16695239 TI - STUDIES ON PAIN: QUANTITATIVE MEASUREMENTS OF TWO PAIN SENSATIONS OF THE SKIN, WITH REFERENCE TO THE NATURE OF THE "HYPERALGESIA OF PERIPHERAL NEURITIS". PMID- 16695240 TI - THE CIRCULATION IN EXPERIMENTAL NEUROGENIC HYPERTENSION. PMID- 16695241 TI - NITROGEN METABOLISM IN ACUTE INFECTIONS. PMID- 16695242 TI - ANTISTREPTOLYSIN TITERS IN CASES OF FILARIASIS WITH RECURRENT LYMPHANGITIS AMONG MILITARY PERSONNEL. PMID- 16695243 TI - PROTECTIVE EFFECT OF VACCINATION AGAINST INDUCED INFLUENZA A. PMID- 16695244 TI - PROTECTIVE EFFECT OF VACCINATION AGAINST INDUCED INFLUENZA B. PMID- 16695245 TI - MATURATION AND DESTRUCTION OF TRANSFUSED HUMAN RETICULOCYTES. EVALUATION OF RETICULOCYTE EXPERIMENTS FOR THE MEASUREMENT OF HEMOGLOBIN METABOLISM. PMID- 16695246 TI - CALCIUM METABOLISM IN NEPHROSIS. I. A DESCRIPTION OF AN ABNORMALITY IN CALCIUM METABOLISM IN CHILDREN WITH NEPHROSIS. PMID- 16695247 TI - STUDIES IN ASCORBIC ACID WITH ESPECIAL REFERENCE TO THE WHITE LAYER. I. DESCRIPTION OF METHOD AND COMPARISON OF ASCORBIC ACID LEVELS IN WHOLE BLOOD, PLASMA, RED CELLS, AND WHITE LAYER. PMID- 16695248 TI - METABOLIC ALTERATIONS FOLLOWING THERMAL BURNS. I. NITROGEN BALANCE IN EXPERIMENTAL BURNS. PMID- 16695249 TI - THE USE OF SODIUM p-AMINOHIPPURATE FOR THE FUNCTIONAL EVALUATION OF THE HUMAN KIDNEY. PMID- 16695250 TI - THE TREATMENT OF LOBAR PNEUMONIA WITH PENICILLIN. PMID- 16695251 TI - THE USE OF PENICILLIN IN THE LOCAL TREATMENT OF PNEUMOCOCCAL EMPYEMA. PMID- 16695252 TI - THE EFFECT OF CONTINUOUS INTRAVENOUS ADMINISTRATION OF HISTAMINE ON BASAL METABOLISM IN HUMAN BEINGS. PMID- 16695253 TI - LIVER FUNCTION TESTS IN CHRONIC RELAPSING VIVAX MALARIA. PMID- 16695254 TI - THE LIPIDS OF SERUM AND LIVER IN PATIENTS WITH HEPATIC DISEASES. PMID- 16695255 TI - ELECTROPHORETIC CHANGES IN THE PLASMA PROTEIN PATTERNS OF PATIENTS WITH RELAPSING MALARIA. PMID- 16695256 TI - ELECTROPHORETIC CHANGES IN THE SERUM PROTEIN PATTERNS OF PATIENTS WITH SCARLET FEVER AND RHEUMATIC FEVER. PMID- 16695257 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XXIV. STUDIES ON THE NUTRITIVE VALUE OF HUMAN PLASMA FRACTIONS. PMID- 16695258 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XXV. THE PREPARATION OF ANTI-A ISOAGGLUTININ REAGENTS FROM MIXED BLOOD OF GROUPS O AND B. PMID- 16695259 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XXVI. THE PROPERTIES OF SOLUTIONS OF HUMAN SERUM ALBUMIN OF LOW SALT CONTENT. PMID- 16695260 TI - THE TREATMENT OF EXPERIMENTAL HYPOPARATHYROIDISM IN DOGS. PMID- 16695261 TI - ELECTROCARDIOGRAMS IN TRAUMATIC SHOCK IN MAN. PMID- 16695262 TI - QUANTITATIVE ELECTROENCEPHALOGRAPHIC STUDIES OF ANOXIA IN HUMANS; COMPARISON WITH ACUTE ALCOHOLIC INTOXICATION AND HYPOGLYCEMIA. PMID- 16695263 TI - STUDIES IN BLOOD COAGULATION: THE COAGULATION PROPERTIES OF CERTAIN GLOBULIN FRACTIONS OF NORMAL HUMAN PLASMA IN VITRO. PMID- 16695264 TI - THE COAGULATION DEFECT IN HEMOPHILIA: THE EFFECT, IN HEMOPHILIA, OF THE PARENTERAL ADMINISTRATION OF A FRACTION OF THE PLASMA GLOBULINS RICH IN FIBRINOGEN. PMID- 16695265 TI - THE INCREASED PLASMA VOLUME IN CARDIAC INSUFFICIENCY: ITS CORRELATION WITH RIGHT SIDED FAILURE. PMID- 16695266 TI - THE SOLAR HEAT LOAD: ITS RELATIONSHIP TO TOTAL HEAT LOAD AND ITS RELATIVE IMPORTANCE IN THE DESIGN OF CLOTHING. PMID- 16695267 TI - SERUM IODINE OF EUTHYROID SUBJECTS TREATED WITH DESICCATED THYROID. PMID- 16695268 TI - SERUM IODINE IN HYPOTHYROIDISM BEFORE AND DURING THYROID THERAPY. PMID- 16695269 TI - TYPE SPECIFIC MENINGOCOCCIC AGGLUTININS: II-THE RELATIONSHIP OF TITERS TO THE COURSE OF THE DISEASE. PMID- 16695270 TI - HEMODYNAMIC ALTERATIONS IN NORMOTENSIVE AND HYPERTENSIVE SUBJECTS DURING THE PYROGENIC REACTION. PMID- 16695271 TI - THE EFFECT OF SALICYLATES ON THE ELECTROLYTE STRUCTURE OF THE BLOOD PLASMA. I. RESPIRATORY ALKALOSIS IN MONKEYS AND DOGS AFTER SODIUM AND METHYL SALICYLATE; THE INFLUENCE OF HYPNOTIC DRUGS AND OF SODIUM BICARBONATE ON SALICYLATE POISONING. PMID- 16695272 TI - THE EFFECT OF SALICYLATES ON THE ELECTROLYTE STRUCTURE OF THE BLOOD PLASMA. II. THE ACTION OF THERAPEUTIC DOSES OF SODIUM SALICYLATE AND OF ACETYLSALICYLIC ACID IN MAN. PMID- 16695273 TI - EPIDEMIC OF ACUTE RESPIRATORY DISEASE ASSOCIATED WITH ATYPICAL PNEUMONIA. PMID- 16695274 TI - THE EFFECT OF ALTERATIONS IN BLOOD VOLUME ON THE ANEMIA AND HYPOPROTEINEMIA OF HUMAN MALARIA. PMID- 16695275 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION XXIX. SERUM ALBUMIN AS A DILUENT FOR Rh TYPING REAGENTS. PMID- 16695276 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION XXX. THE USE OF SALT-POOR CONCENTRATED HUMAN SERUM ALBUMIN SOLUTION IN THE TREATMENT OF CHRONIC BRIGHT'S DISEASE. PMID- 16695277 TI - THE TOXIC FACTORS IN EXPERIMENTAL TRAUMATIC SHOCK. I. PHYSIOLOGIC EFFECTS OF MUSCLE LIGATION IN THE DOG. PMID- 16695278 TI - THE TOXIC FACTORS IN EXPERIMENTAL TRAUMATIC SHOCK. II. STUDIES ON ELECTROLYTE AND WATER BALANCE IN SHOCK. PMID- 16695279 TI - THE TOXIC FACTORS IN EXPERIMENTAL TRAUMATIC SHOCK. III. SHOCK ACCOMPANYING MUSCLE ISCHEMIA AND LOSS OF VASCULAR FLUID. PMID- 16695280 TI - THE TOXIC FACTORS IN EXPERIMENTAL TRAUMATIC SHOCK. IV. THE EFFECTS OF THE INTRAVENOUS INJECTION OF THE EFFUSION FROM ISCHEMIC MUSCLE. PMID- 16695281 TI - THE TOXIC FACTORS IN EXPERIMENTAL TRAUMATIC SHOCK. V. CHEMICAL AND ENZYMATIC PROPERTIES OF MUSCLE EXUDATE. PMID- 16695282 TI - THE TOXIC FACTORS IN EXERIMENTAL TRAUMATIC SHOCK. VI. THE TOXIC INFLUENCE OF THE BACTERIAL FLORA, PARTICULARLY CLOSTRIDIUM WELCHII, IN EXUDATES OF ISCHEMIC MUSCLE. PMID- 16695283 TI - SOME OBSERVATIONS ON THE EFFECT OF INJECTED CYTOCHROME C IN ANIMALS. PMID- 16695284 TI - EFFECT OF IODINE ON THE THYROID GLAND IN GRAVES' DISEASE WHEN GIVEN IN CONJUNCTION WITH THIOURACIL-A TWO-ACTION THEORY OF IODINE. PMID- 16695285 TI - CIRCULATION AND RESPIRATION DURING AN EPISODE OF CHILL AND FEVER IN MAN. PMID- 16695286 TI - THE ESTIMATION OF HEPATIC BLOOD FLOW IN MAN. PMID- 16695287 TI - THE ABSORPTION AND EXCRETION OF STREPTOMYCIN IN HUMAN CHRONIC TYPHOID CARRIERS. PMID- 16695288 TI - TRAUMATIC SHOCK. XII. HEMODYNAMIC EFFECTS OF ALTERATIONS OF BLOOD VISCOSITY IN NORMAL DOGS AND IN DOGS IN SHOCK. PMID- 16695289 TI - "BACILLUS MUCOSUS CAPSULATUS" IN INFANTILE DIARRHEA. PMID- 16695290 TI - THE EFFECTS OF ACTIVE AND PASSIVE HYPERVENTILATION ON CEREBRAL BLOOD FLOW, CEREBRAL OXYGEN CONSUMPTION, CARDIAC OUTPUT, AND BLOOD PRESSURE OF NORMAL YOUNG MEN. PMID- 16695291 TI - HEMODYNAMIC CHANGES IN SALT DEPLETION AND IN DEHYDRATION. PMID- 16695292 TI - THE TREATMENT OF SHOCK DUE TO SALT DEPLETION; COMPARISON OF THE HEMODYNAMIC EFFECTS OF ISOTONIC SALINE, OF HYPERTONIC SALINE, AND OF ISOTONIC GLUCOSE SOLUTIONS. PMID- 16695293 TI - THE RELATIONSHIPS OF OXYGEN, CARBON DIOXIDE, AND HEMOGLOBIN IN THE BLOOD OF MAN: OXYHEMOGLOBIN DISSOCIATION UNDER VARIOUS PHYSIOLOGICAL CONDITIONS. PMID- 16695294 TI - HEPATITIS WITHOUT JAUNDICE IN INFECTIOUS MONONUCLEOSIS. PMID- 16695295 TI - TRAUMATIC SHOCK. XIII. THE PREVENTION OF IRREVERSIBILITY IN HEMORRHAGIC SHOCK BY VIVIPERFUSION OF THE LIVER. PMID- 16695296 TI - HEMORRHAGIC SHOCK: DEFINITION AND CRITERIA FOR ITS DIAGNOSIS. PMID- 16695297 TI - PHOTOELECTRIC STUDY OF SOME FACTORS RELATED TO BLOOD CLOTTING. PMID- 16695298 TI - INFECTIOUS HEPATITIS COMPLICATED BY SECONDARY INVASION WITH SALMONELLA. PMID- 16695299 TI - ON THE CAUSE OF THE RESPIRATORY VARIATION OF THE BALLISTOCARDIOGRAM, WITH A NOTE ON SINUS ARRHYTHMIA. PMID- 16695300 TI - THE ANEMIA OF INFECTION. I. HYPOFERREMIA, HYPERCUPREMIA, AND ALTERATIONS IN PORPHYRIN METABOLISM IN PATIENTS. PMID- 16695301 TI - THE ANEMIA OF INFECTION. II. THE EXPERIMENTAL PRODUCTION OF HYPOFERREMIA AND ANEMIA IN DOGS. PMID- 16695302 TI - THE ABSORPTION INTO AND DISTRIBUTION OF PENICILLIN IN THE CEREBROSPINAL FLUID. PMID- 16695303 TI - THE ABSORPTION, DISTRIBUTION, AND EXCRETION OF STREPTOMYCIN IN MAN. PMID- 16695304 TI - FAILURE OF LOCAL FLUID LOSS TO ACCOUNT FOR DEATH IN EXPERIMENTAL SHOCK. PMID- 16695305 TI - BLOOD VOLUME IN CLINICAL SHOCK. I. MIXING TIME AND DISAPPEARANCE RATE OF T-1824 IN NORMAL SUBJECTS AND IN PATIENTS IN SHOCK; DETERMINATION OF PLASMA VOLUME IN MAN FROM 10-MINUTE SAMPLE. PMID- 16695307 TI - EFFECT OF ADMINISTRATION OF ESSENTIAL AMINO ACIDS ON UTILIZATION OF DIETARY PROTEIN IN MALNUTRITION AMONG WAR CASUALTIES. PMID- 16695306 TI - BLOOD VOLUME IN CLINICAL SHOCK. II. THE EXTENT AND CAUSE OF BLOOD VOLUME REDUCTION IN TRAUMATIC, HEMORRHAGIC, AND BURN SHOCK. PMID- 16695308 TI - THE ABSORPTION, EXCRETION, AND DESTRUCTION OF ORALLY ADMINISTERED PENICILLIN. PMID- 16695309 TI - TREATMENT OF EXPERIMENTAL UREMIA BY MEANS OF PERITONEAL IRRIGATION. PMID- 16695310 TI - THE ROLE OF COLLOID AND OF SALINE IN THE TREATMENT OF SHOCK. PMID- 16695311 TI - A COMPARISON OF THE EFFECTS OF HIGH AND LOW VISCOSITY GELATINS AFTER THEIR INTRAVENOUS INJECTION IN MAN. PMID- 16695312 TI - CENTRAL VENOUS PRESSURE IN RELATION TO CARDIAC "COMPETENCE," BLOOD VOLUME AND EXERCISE. PMID- 16695313 TI - CHLORIDE EXCRETION DURING GLYCOSURIA IN PATIENTS WITH DIABETES. PMID- 16695314 TI - ESTIMATION OF BLOOD VOLUME FROM CHANGE IN BLOOD SPECIFIC GRAVITY FOLLOWING A PLASMA INFUSION. PMID- 16695315 TI - THE ISOLATION OF POLIOMYELITIS VIRUS FROM HUMAN EXTRA-NEURAL SOURCES. I. COMPARISON OF VIRUS CONTENT OF PHARYNGEAL SWABS, OROPHARYNGEAL WASHINGS, AND STOOLS OF PATIENTS. PMID- 16695316 TI - THE ISOLATION OF POLIOMYELITIS VIRUS FROM HUMAN EXTRA-NEURAL SOURCES. II. COMPARISON OF VIRUS CONTENT OF BLOOD, OROPHARYNGEAL WASHINGS, AND STOOLS OF CONTACTS. PMID- 16695317 TI - THE ISOLATION OF POLIOMYELITIS VIRUS FROM HUMAN EXTRA-NEURAL SOURCES. III. PERSISTENCE OF VIRUS IN STOOLS AFTER ACUTE INFECTION. PMID- 16695318 TI - THE ISOLATION OF POLIOMYELITIS VIRUS FROM HUMAN EXTRA-NEURAL SOURCES. IV. SEARCH FOR VIRUS IN THE BLOOD OF PATIENTS. PMID- 16695319 TI - STUDIES OF THIAMINE METABOLISM IN MAN. I. THIAMINE BALANCE. THE NORMAL REQUIREMENT OF VITAMIN B(1) AND THE ROLE OF FECAL THIAMINE IN HUMAN NUTRITION. PMID- 16695320 TI - STUDIES OF THIAMINE METABOLISM IN MAN. II. THIAMINE AND PYRIMIDINE EXCRETION WITH SPECIAL REFERENCE TO THE RELATIONSHIP BETWEEN INJECTED AND EXCRETED THIAMINE IN NORMAL AND ABNORMAL SUBJECTS. PMID- 16695321 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XXXI. THE USE OF SALT-POOR CONCENTRATED HUMAN SERUM ALBUMIN SOLUTION IN THE TREATMENT OF HEPATIC CIRRHOSIS. PMID- 16695322 TI - CHANGES IN MUSCLE COMPOSITION IN ALKALOSIS. PMID- 16695323 TI - STUDIES OF STREPTOCOCCAL FIBRINOLYSIS. I. THE DISSIMILARITY OF SERUM PROTEASE AND TRYPSIN AS INDICATED BY THE SEPARATE SPECIFICITIES OF THEIR KINASES, FIBRINOLYSIN AND ENTEROKINASE. PMID- 16695324 TI - STUDIES OF STREPTOCOCCAL FIBRINOLYSIS. II. THE INHIBITION OF STREPTOCOCCAL FIBRINOLYSIS BY ANTIFIBRINOLYSIN AND ANTIPROTEASE. PMID- 16695325 TI - STUDIES OF STREPTOCOCCAL FIBRINOLYSIS. III. A QUANTITATIVE METHOD FOR THE ESTIMATION OF SERUM ANTIFIBRINOLYSIN. PMID- 16695326 TI - STUDIES OF STREPTOCOCCAL FIBRINOLYSIS. IV. CLINICAL APPLICATION OF A QUANTITATIVE ANTIFIBRINOLYSIN TEST. PMID- 16695327 TI - EFFECTS OF THYROID ON CREATINE METABOLISM WITH A DISCUSSION OF THE MECHANISM OF STORAGE AND EXCRETION OF CREATINE BODIES. PMID- 16695328 TI - A SURVEY OF THE TWENTY-FOUR-HOUR URIC ACID AND UREA CLEARANCES IN ECLAMPSIA AND SEVERE PREECLAMPSIA. PMID- 16695329 TI - ON THE INCREASED URIC ACID CLEARANCE FOLLOWING THE INTRAVENOUS INFUSION OF HYPERTONIC GLUCOSE SOLUTIONS. PMID- 16695330 TI - EDEMA AND DECREASED RENAL BLOOD FLOW IN PATIENTS WITH CHRONIC CONGESTIVE HEART FAILURE: EVIDENCE OF "FORWARD FAILURE" AS THE PRIMARY CAUSE OF EDEMA. PMID- 16695331 TI - THE EFFECT OF SIGNIFICANT WEIGHT CHANGE ON THE PREDICTED PLASMA VOLUME. PMID- 16695332 TI - SERUM IODINE IN HYPERTHYROIDISM, WITH PARTICULAR REFERENCE TO THE EFFECTS OF SUBTOTAL THYROIDECTOMY. PMID- 16695333 TI - BALLISTOCARDIOGRAPHIC STUDY OF CHANGES IN CARDIAC OUTPUT DUE TO RESPIRATION. PMID- 16695334 TI - PROPERTIES OF AN ANTICOAGULANT FOUND IN THE BLOOD OF A HEMOPHILIAC. PMID- 16695335 TI - STUDIES IN ASCORBIC ACID WITH ESPECIAL REFERENCE TO THE WHITE LAYER. II. THE RELATION OF INTAKE TO BLOOD LEVELS IN NORMAL CHILDREN AND THE EFFECT OF ACUTE AND CHRONIC ILLNESS. PMID- 16695336 TI - THE EFFECT OF CLOTHING ON THE ABILITY OF MEN TO WORK IN INTENSE HEAT. PMID- 16695337 TI - AN OBJECTIVE METHOD FOR DETERMINING CIRCULATION TIME FROM PULMONARY TO SYSTEMIC CAPILLARIES BY THE USE OF THE OXIMETER. PMID- 16695339 TI - CLINICAL USES OF 2,3-DIMERCAPTOPROPANOL (BAL). II. THE EFFECT OF BAL ON THE EXCRETION OF ARSENIC IN NORMAL SUBJECTS AND AFTER MINIMAL EXPOSURE TO ARSENICAL SMOKE. PMID- 16695340 TI - CLINICAL USES OF 2,3-DIMERCAPTOPROPANOL (BAL). III. STUDIES ON THE TOXICITY OF BAL ON PERCUTANEOUS AND PARENTERAL ADMINISTRATION. PMID- 16695341 TI - CLINICAL USES OF 2,3-DIMERCAPTOPROPANOL (BAL). IV. PHARMACOLOGIC OBSERVATIONS ON BAL BY INTRAMUSCULAR INJECTION IN MAN. PMID- 16695342 TI - CLINICAL USES OF 2,3-DIMERCAPTOPROPANOL (BAL). V. SKIN SENSITIZATION TO BAL. PMID- 16695344 TI - CLINICAL USES OF 2,3-DIMERCAPTOPROPANOL (BAL). VII. THE TREATMENT OF ARSENICAL DERMATITIS WITH PREPARATIONS OF BAL. PMID- 16695345 TI - CLINICAL USES OF 2,3-DIMERCAPTOPROPANOL (BAL). VIII. THE EFFECT OF BAL ON THE EXCRETION OF ARSENIC IN ARSENICAL INTOXICATION. PMID- 16695347 TI - CLINICAL USES OF 2,3-DIMERCAPTOPROPANOL (BAL). X. THE TREATMENT OF ACUTE SYSTEMIC MERCURY POISONING IN EXPERIMENTAL ANIMALS WITH BAL, THIOSORBITOL AND BAL GLUCOSIDE. PMID- 16695348 TI - CLINICAL USES OF 2,3-DIMERCAPTOPROPANOL (BAL). XI. THE TREATMENT OF ACUTE MERCURY POISONING BY BAL. PMID- 16695349 TI - KIDNEY FUNCTION AND CIRCULATORY COLLAPSE. POST-SYNCOPAL OLIGURIA. PMID- 16695350 TI - THE FATE OF INTRAVENOUSLY INJECTED GELATIN IN HUMAN SUBJECTS. PMID- 16695351 TI - CHANGES IN BLOOD COAGULATION FOLLOWING CORONARY THROMBOSIS MEASURED BY THE HEPARIN RETARDED CLOTTING TEST (WAUGH AND RUDDICK TEST). PMID- 16695352 TI - ADDITIVE EFFECTS OF IODINE AND THIOUREA IN THE TREATMENT OF HYPERTHYROIDISM. PMID- 16695353 TI - THE USE OF TWO RADIOACTIVE ISOTOPES OF IRON IN TRACER STUDIES OF ERYTHROCYTES. PMID- 16695354 TI - THE MEASUREMENT OF THE CIRCULATING RED CELL VOLUME BY MEANS OF TWO RADIOACTIVE ISOTOPES OF IRON. PMID- 16695356 TI - RECORDING OF RIGHT HEART PRESSURES IN NORMAL SUBJECTS AND IN PATIENTS WITH CHRONIC PULMONARY DISEASE AND VARIOUS TYPES OF CARDIO-CIRCULATORY DISEASE. PMID- 16695355 TI - A CLINICAL STUDY OF TRANSFUSION REACTIONS: THE HEMOLYTIC EFFECT OF GROUP-O BLOOD AND POOLED PLASMA CONTAINING INCOMPATIBLE ISOAGGLUTININS. PMID- 16695357 TI - STUDIES ON BACTERIA DEVELOPING RESISTANCE TO PENICILLIN FRACTIONS X AND G IN VITRO AND IN PATIENTS UNDER TREATMENT FOR BACTERIAL ENDOCARDITIS. PMID- 16695358 TI - THE EFFECT OF INFECTION AND TRAUMA ON THE EXCRETION OF URINARY CORTIN. PMID- 16695359 TI - PARENTERAL NUTRITION. II. THE UTILIZATION OF EMULSIFIED FAT GIVEN INTRAVENOUSLY. PMID- 16695360 TI - AN HISTOLOGIC STUDY OF THE EFFECTS OF EXPERIMENTAL BOTULINUS POISONING ON THE LIVERS OF GUINEA PIGS. PMID- 16695361 TI - THE INFLUENCE OF AMMONIUM ION ON THE PLASMA ATABRINE LEVEL AND ON THE URINARY EXCRETION OF ATABRINE. PMID- 16695362 TI - PLASMA alpha-AMINO ACID NITROGEN AND SERUM LIPIDS OF SURGICAL PATIENTS. PMID- 16695363 TI - SOME OBSERVATIONS ON MEN SITTING QUIETLY IN EXTREME COLD. PMID- 16695364 TI - EFFECT OF METHEMOGLOBINEMIA ON THE VISUAL THRESHOLD AT SEA LEVEL, AT HIGH ALTITUDES, AND AFTER EXERCISE. PMID- 16695365 TI - VOLUNTARY BREATHHOLDING. I. PULMONARY GAS EXCHANGE DURING BREATHHOLDING. PMID- 16695366 TI - VOLUNTARY BREATHHOLDING. II. THE RELATION OF THE MAXIMUM TIME OF BREATHHOLDING TO THE OXYGEN TENSION OF THE INSPIRED AIR. PMID- 16695367 TI - VOLUNTARY BREATHHOLDING. III. THE RELATION OF THE MAXIMUM TIME OF BREATHHOLDING TO THE OXYGEN AND CARBON DIOXIDE TENSIONS OF ARTERIAL BLOOD, WITH A NOTE ON ITS CLINICAL AND PHYSIOLOGICAL SIGNIFICANCE. PMID- 16695368 TI - FLUID LOSS IN RATS WITH TOURNIQUET SHOCK. PMID- 16695369 TI - STUDIES ON LEPTOSPIRA ICTEROHAEMORRHAGIAE. II. A CRITICAL STUDY OF THE EFFECT OF PENICILLIN ON LEPTOSPIRA ICTEROHAEMORRHAGIAE IN VITRO AND IN LEPTOSPIROSIS IN GUINEA PIGS. PMID- 16695370 TI - QUANTITATION AND REGIONAL DISTRIBUTION OF SWEAT GLANDS IN MAN. PMID- 16695371 TI - GALACTOSE DISAPPEARANCE FROM THE BLOOD STREAM. CALCULATION OF A GALACTOSE REMOVAL CONSTANT AND ITS APPLICATION AS A TEST FOR LIVER FUNCTION. PMID- 16695372 TI - DETERMINATION OF THE RATE OF DISAPPEARANCE OF MUSTARD GAS AND MUSTARD INTERMEDIATES IN CORNEAL TISSUE. PMID- 16695373 TI - DEMONSTRATION OF A TOXIC FACTOR IN THE BLOOD OF RATS SHOCKED BY BURN. PMID- 16695374 TI - THE ACTION OF HISTAMINE ON THE RESPIRATORY TRACT IN NORMAL AND ASTHMATIC SUBJECTS. PMID- 16695375 TI - THE EFFECT OF ANTIHISTAMINE SUBSTANCES AND OTHER DRUGS ON HISTAMINE BRONCHOCONSTRICTION IN ASTHMATIC SUBJECTS. PMID- 16695376 TI - COMPLEMENT IN INFECTIOUS DISEASE IN MAN. PMID- 16695377 TI - COMPLEMENT AND ISOHEMAGGLUTININS IN URINARY PROTEINS. PMID- 16695378 TI - ELECTROPHORETIC ISOLATION OF A CIRCULATING ANTICOAGULANT. PMID- 16695379 TI - A COMPARISON OF THE CEPHALIN-CHOLESTEROL FLOCCULATION AND THYMOL TURBIDITY TESTS IN PATIENTS WITH EXPERIMENTALLY INDUCED INFECTIOUS HEPATITIS. PMID- 16695380 TI - METABOLIC STUDIES IN LOUSE-BORNE TYPHUS. OBSERVATIONS ON SERUM ELECTROLYTE PATTERN, SERUM PROTEIN PARTITION, AND NITROGEN BALANCE. PMID- 16695381 TI - CIRCULATING RED CELL VOLUME MEASURED SIMULTANEOUSLY BY THE RADIOACTIVE IRON AND DYE METHODS. PMID- 16695382 TI - THE DISTRIBUTION OF RED CELLS AND PLASMA IN LARGE AND MINUTE VESSELS OF THE NORMAL DOG, DETERMINED BY RADIOACTIVE ISOTOPES OF IRON AND IODINE. PMID- 16695383 TI - COMPARISON OF THE EFFECTS OF MASSIVE BLOOD TRANSFUSIONS AND OF LIVER EXTRACT IN PERNICIOUS ANEMIA. PMID- 16695384 TI - CHEMICAL, CLINICAL AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION: XXXII. THE COAGULATION DEFECT IN HEMOPHILIA. AN IN VITRO AND IN VIVO COMPARISON OF NORMAL AND HEMOPHILIC WHOLE BLOOD, PLASMA AND DERIVED PLASMA PROTEIN FRACTIONS. PMID- 16695385 TI - CHEMICAL, CLINICAL AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XXXIII. THE COAGULATION DEFECT IN HEMOPHILIA: THE EFFECT IN VITRO AND IN VIVO ON THE COAGULATION TIME IN HEMOPHILIA OF A PROTHROMBIN AND FIBRINOGEN FREE NORMAL PLASMA AND ITS DERIVED PROTEIN FRACTIONS. PMID- 16695386 TI - STUDIES ON HYPERVENTILATION. II. ELECTROCARDIOGRAPHIC CHANGES IN NORMAL MAN DURING VOLUNTARY HYPERVENTILATION. PMID- 16695387 TI - MEASUREMENTS RELATED TO PAIN IN NEUROCIRCULATORY ASTHENIA, ANXIETY NEUROSIS, OR EFFORT SYNDROME: LEVELS OF HEAT STIMULUS PERCEIVED AS PAINFUL AND PRODUCING WINCE AND WITHDRAWAL REACTIONS. PMID- 16695388 TI - RENAL REABSORPTION OF CHLORIDE AND PHOSPHATE IN NORMAL SUBJECTS AND IN PATIENTS WITH ESSENTIAL ARTERIAL HYPERTENSION. PMID- 16695389 TI - THE CARDIAC OUTPUT IN MAN: STUDIES WITH THE LOW FREQUENCY, CRITICALLY-DAMPED BALLISTOCARDIOGRAPH, AND THE METHOD OF RIGHT ATRIAL CATHETERIZATION. PMID- 16695390 TI - THE ANEMIA OF INFECTION. III. THE UPTAKE OF RADIO-ACTIVE IRON IN IRON-DEFICIENT AND IN PYRIDOXINE-DEFICIENT PIGS BEFORE AND AFTER ACUTE INFLAMMATION. PMID- 16695391 TI - INTUBATION STUDIES OF THE HUMAN SMALL INTESTINE. XXV. THE ABSORPTION OF GALACTOSE FROM THE INTESTINE OF NORMAL INDIVIDUALS AND THYROTOXIC PATIENTS. PMID- 16695392 TI - THE ANEMIA OF INFECTION. IV. THE LACK OF RELATIONSHIP BETWEEN THE DIVERSION OF IRON FROM THE PLASMA AND THE ORIGIN OF THE ANEMIA. PMID- 16695393 TI - THE ANEMIA OF INFECTION. V. FATE OF INJECTED RADIOACTIVE IRON IN THE PRESENCE OF INFLAMMATION. PMID- 16695394 TI - THE CIRCULATING RED CELL AND PLASMA VOLUME AND THE DISTRIBUTION OF BLOOD IN LARGE AND MINUTE VESSELS IN EXPERIMENTAL SHOCK IN DOGS, MEASURED BY RADIOACTIVE ISOTOPES OF IRON AND IODINE. PMID- 16695395 TI - THE NUTRITIONAL REQUIREMENTS FOR NITROGEN BALANCE IN SURGICAL PATIENTS DURING THE EARLY POSTOPERATIVE PERIOD. PMID- 16695396 TI - THE METABOLIC EFFECTS OF STEROID HORMONES IN OSTEOPOROSIS. PMID- 16695397 TI - IMMUNOLOGIC STUDIES IN INSULIN RESISTANCE. III. MEASUREMENT OF AN INSULIN ANTAGONIST IN THE SERUM OF AN INSULIN-RESISTANT PATIENT BY THE BLOOD SUGAR CURVE METHOD IN MICE. PMID- 16695398 TI - TRICHINELLA SKIN TESTS IN AN ORPHANAGE AND PRISON. COMPARISON WITH SEROLOGIC TESTS FOR TRICHINOSIS AND WITH THE TUBERCULIN REACTION. PMID- 16695399 TI - THE EFFECT ON THE TRICHINELLA SKIN TEST OF SIMULTANEOUS INFECTION WITH BOVINE TUBERCULOSIS IN TRICHINOUS RABBITS. PMID- 16695400 TI - THE EFFECT OF PEPTIC AND TRYPTIC DIGESTION ON THE ANTIGENICITY OF TRICHINELLA SPIRALIS. PMID- 16695401 TI - PAMAQUINE NAPHTHOATE AS A PROPHYLACTIC FOR MALARIAL INFECTIONS. PMID- 16695402 TI - ELECTROPHORETIC CHANGES IN THE SERUM OF A PATIENT WITH RHEUMATOID ARTHRITIS. PMID- 16695403 TI - VARIATION IN PROTEIN AND POLYSACCHARIDE CONTENT OF SERA IN THE CHRONIC DISEASES, TUBERCULOSIS, SARCOIDOSIS, AND CARCINOMA. PMID- 16695404 TI - A STUDY OF THE RESIDUAL EFFECTS OF PHOSGENE POISONING IN HUMAN SUBJECTS. I. AFTER ACUTE EXPOSURE. PMID- 16695405 TI - A STUDY OF THE RESIDUAL EFFECTS OF PHOSGENE POISONING IN HUMAN SUBJECTS. II. AFTER CHRONIC EXPOSURE. PMID- 16695406 TI - FACTORS INFLUENCING THE URINARY EXCRETION OF CALCIUM. I. IN NORMAL PERSONS. PMID- 16695407 TI - DIFFERENTIAL SPINAL BLOCK. II. THE REACTION OF SUDOMOTOR AND VASOMOTOR FIBERS. PMID- 16695408 TI - SERUM POTASSIUM, MAGNESIUM, AND CALCIUM LEVELS IN DIABETIC ACIDOSIS. PMID- 16695409 TI - STUDIES ON GANGRENE FOLLOWING COLD INJURY. I. A METHOD FOR PRODUCING GANGRENE BY MEANS OF CONTROLLED INJURY BY COLD. PMID- 16695410 TI - STUDIES ON GANGRENE FOLLOWING COLD INJURY. II. GENERAL COURSE OF EVENTS IN RABBIT FEET AND EARS FOLLOWING UNTREATED COLD INJURY. PMID- 16695411 TI - STUDIES ON GANGRENE FOLLOWING COLD INJURY. III. EDEMA FOLLOWING COLD INJURY: ITS MAGNITUDE AND THE COMPOSITION AND SOURCES OF EDEMA FLUID. PMID- 16695412 TI - STUDIES ON GANGRENE FOLLOWING COLD INJURY. IV. THE USE OF FLUORESCEIN AS AN INDICATOR OF LOCAL BLOOD FLOW: DISTRIBUTION OF FLUORESCEIN IN BODY FLUIDS AFTER INTRAVENOUS INJECTION. PMID- 16695413 TI - STUDIES ON GANGRENE FOLLOWING COLD INJURY. V. THE USE OF FLUORESCEIN AS AN INDICATOR OF LOCAL BLOOD FLOW: FLUORESCEIN TESTS IN EXPERIMENTAL FROSTBITE. PMID- 16695414 TI - EFFECT OF CREATINE ON THE PAIN OF MUSCLE ISCHEMIA. PMID- 16695415 TI - SCLEROMA: AN ETIOLOGICAL STUDY. PMID- 16695416 TI - STUDIES ON A VIRUS FROM A PATIENT WITH FORT BRAGG FEVER (PRETIBIAL FEVER). PMID- 16695417 TI - ELECTROPHORETIC CHANGES IN THE SERUM PROTEIN PATTERN OF A PATIENT WITH TYPHUS FEVER. PMID- 16695418 TI - THE ABSORPTION, EXCRETION, AND PHYSIOLOGICAL EFFECT OF IODINE IN NORMAL HUMAN SUBJECTS. PMID- 16695419 TI - OBSERVATIONS ON MEN PERFORMING A STANDARD AMOUNT OF WORK IN LOW AMBIENT TEMPERATURES. PMID- 16695420 TI - CIRCULATORY DYNAMICS IN THE BASAL STATE OBSERVED DURING CONVALESCENCE. CHANGES IN BODY WEIGHT, BLOOD VOLUME, AND VENOUS PRESSURE. PMID- 16695421 TI - SULFADIAZINE RESISTANT STREPTOCOCCAL INFECTIONS IN A CIVILIAN COMMUNITY. PMID- 16695422 TI - STUDIES ON THE RELATIONSHIP OF THE HEPATITIS VIRUS TO PERSISTENT SYMPTOMS, DISABILITY, AND HEPATIC DISTURBANCE ("CHRONIC HEPATITIS SYNDROME") FOLLOWING ACUTE INFECTIOUS HEPATITIS. PMID- 16695423 TI - BLOOD LACTATE RESPONSE DURING MODERATE EXERCISE IN NEUROCIRCULATORY ASTHENIA, ANXIETY NEUROSIS, OR EFFORT SYNDROME. PMID- 16695424 TI - WATER STORAGE AND THE MOVEMENTS OF BODY FLUIDS AND CHLORIDES DURING ACUTE LIVER DISEASE. PMID- 16695425 TI - THE DISTRIBUTION OF ASCORBIC ACID IN THE BLOOD. PMID- 16695426 TI - FACTORS CONCERNED IN THE CIRCULATORY FAILURE OF ADRENAL INSUFFICIENCY. PMID- 16695427 TI - EFFECT OF BLOOD PROTEASE AND TRYPSIN INHIBITOR ON THE CLOTTING MECHANISM. PMID- 16695428 TI - A STUDY OF THE DISTRIBUTION AND FATE OF ANTIMONY WHEN USED AS TARTAR EMETIC AND FUADIN IN THE TREATMENT OF AMERICAN SOLDIERS WITH SCHISTOSOMIASIS JAPONICA. PMID- 16695429 TI - PENICILLIN-RESISTANT STAPHYLOCOCCI: MECHANISMS INVOLVED IN THE DEVELOPMENT OF RESISTANCE. PMID- 16695430 TI - PHYSIOLOGIC ACTION OF CLOSTRIDIUM WELCHII (TYPE A) TOXINS IN DOGS. PMID- 16695431 TI - PHYSIOLOGIC ACTION OF CLOSTRIDIUM OEDEMATIENS (NOVYI) TOXIN IN DOGS. PMID- 16695432 TI - THE INFLUENCE OF ANTITOXIN UPON THE ACTION OF CLOSTRIDIUM OEDEMATIENS TOXIN IN THE HEART-LUNG PREPARATION OF THE DOG. PMID- 16695433 TI - THE SCATTER AND ABSORPTION OF LIGHT BY THE EXCISED CORNEA AT VARIOUS WAVELENGTHS DETERMINED BY A PHOTOELECTRIC METHOD. THE EFFECT OF TRAUMA AND VACCINIAL INFECTION. PMID- 16695434 TI - SEROLOGICAL STUDIES ON INFLUENZA DURING A NINE-MONTH PERIOD. PMID- 16695435 TI - COMPARATIVE ACTION OF ACETYL-BETA-METHYL CHOLINE AND HISTAMINE ON THE RESPIRATORY TRACT IN NORMALS, PATIENTS WITH HAY FEVER, AND SUBJECTS WITH BRONCHIAL ASTHMA. PMID- 16695436 TI - THE EFFECT OF ORALLY AND INTRAVENOUSLY ADMINISTERED AMINO ACID MIXTURES ON VOLUNTARY FOOD CONSUMPTION IN NORMAL MEN. PMID- 16695437 TI - MINIMUM DOSAGE OF THIOUREA, GIVEN TOGETHER WITH IODINE MEDICATION, NECESSARY FOR THE PRODUCTION AND MAINTENANCE OF A REMISSION IN HYPERTHYROIDISM. PMID- 16695438 TI - VASODILATOR FIBERS IN THE HUMAN SKIN. PMID- 16695439 TI - THE EFFECT OF VARYING PULMONARY PRESSURE ON THE ARTERIAL PRESSURES IN MAN AND ANESTHETIZED CATS. PMID- 16695440 TI - STUDIES ON GANGRENE FOLLOWING COLD INJURY: VI. CAPILLARY BLOOD FLOW AFTER COLD INJURY, THE EFFECTS OF RAPID WARMING, AND SYMPATHETIC BLOCK. PMID- 16695441 TI - STUDIES ON GANGRENE FOLLOWING COLD INJURY. VII. TREATMENT OF COLD INJURY BY MEANS OF IMMEDIATE RAPID WARMING. PMID- 16695442 TI - STUDIES ON GANGRENE FOLLOWING COLD INJURY. VIII. THE USE OF CASTS AND PRESSURE DRESSINGS IN THE TREATMENT OF SEVERE FROSTBITE. PMID- 16695443 TI - STUDIES OF THERMAL INJURY. VI. HYPERPOTASSEMIA CAUSED BY CUTANEOUS EXPOSURE TO EXCESSIVE HEAT. PMID- 16695444 TI - STUDIES OF THERMAL INJURY. VII. PHYSIOLOGICAL MECHANISMS RESPONSIBLE FOR DEATH DURING CUTANEOUS EXPOSURE TO EXCESSIVE HEAT. PMID- 16695445 TI - STUDIES OF BREATHING, PULMONARY VENTILATION AND SUBJECTIVE AWARENESS OF SHORTNESS OF BREATH (DYSPNEA) IN NEUROCIRCULATORY ASTHENIA, EFFORT SYNDROME, ANXIETY NEUROSIS. PMID- 16695446 TI - TRAUMATIC SHOCK. XIV. THE SUCCESSFUL TREATMENT OF HEMORRHAGIC SHOCK BY VIVI PERFUSION OF THE LIVER IN DOGS IRREVERSIBLE TO TRANSFUSION. PMID- 16695447 TI - TRAUMATIC SHOCK. XV. CARBOHYDRATE METABOLISM IN HEMORRHAGIC SHOCK IN THE DOG. PMID- 16695448 TI - STUDIES OF CONGENITAL HEART DISEASE. I. TECHNIQUE OF VENOUS CATHETERIZATION AS A DIAGNOSTIC PROCEDURE. PMID- 16695449 TI - STUDIES OF CONGENITAL HEART DISEASE. II. THE PRESSURE AND OXYGEN CONTENT OF BLOOD IN THE RIGHT AURICLE, RIGHT VENTRICLE, AND PULMONARY ARTERY IN CONTROL PATIENTS, WITH OBSERVATIONS ON THE OXYGEN SATURATION AND SOURCE OF PULMONARY "CAPILLARY" BLOOD. PMID- 16695450 TI - STUDIES OF CONGENITAL HEART DISEASE. III. VENOUS CATHETERIZATION AS A DIAGNOSTIC AID IN PATENT DUCTUS ARTERIOSUS, TETRALOGY OF FALLOT, VENTRICULAR SEPTAL DEFECT, AND AURICULAR SEPTAL DEFECT. PMID- 16695451 TI - A BACTERIOLOGIC STUDY OF THE FACTORS AFFECTING THE EFFICACY OF STREPTOMYCIN THERAPY OF URINARY TRACT INFECTIONS. PMID- 16695452 TI - DIMENSIONAL, OSMOTIC, AND CHEMICAL CHANGES OF ERYTHROCYTES IN STORED BLOOD. I. BLOOD PRESERVED IN SODIUM CITRATE, NEUTRAL, AND ACID CITRATE-GLUCOSE (ACD) MIXTURES. PMID- 16695453 TI - DIMENSIONAL, OSMOTIC, AND CHEMICAL CHANGES OF ERYTHROCYTES IN STORED BLOOD. II. EVALUATION OF SEVERAL ACID AND NEUTRAL PRESERVATION MIXTURES. EFFECT OF STORAGE AT 25 degrees C. IN ALSEVER'S SOLUTION. PMID- 16695454 TI - DIMENSIONAL, OSMOTIC, AND CHEMICAL CHANGES OF ERYTHROCYTES IN STORED BLOOD. III. COMPARISON OF 3 DILUTIONS OF ACID CITRATE-GLUCOSE SOLUTION (ACD). PMID- 16695455 TI - DIMENSIONAL, OSMOTIC, AND CHEMICAL CHANGES OF ERYTHROCYTES IN STORED BLOOD. IV. CELLS SEPARATED FROM PLASMA. PMID- 16695456 TI - THE OSMOTIC RESISTANCE (FRAGILITY) OF HUMAN RED CELLS. PMID- 16695457 TI - WHOLE BLOOD PRESERVATION; A PROBLEM IN GENERAL PHYSIOLOGY. AN IN VITRO ANALYSIS OF THE PROBLEM OF BLOOD STORAGE. PMID- 16695458 TI - ESTIMATION OF CELL SURVIVAL AFTER TRANSFUSION BY SELECTIVE AGGLUTINATION. PMID- 16695459 TI - THE PRESERVATION OF WHOLE BLOOD. PMID- 16695460 TI - THE PRESERVATION OF ERYTHROCYTES LEFT OVER FROM PLASMA PREPARATION. PMID- 16695461 TI - AN ACID CITRATE-DEXTROSE SOLUTION WITH LOW WATER VOLUME AND LOW DEXTROSE CONCENTRATION. PMID- 16695462 TI - THE IN VITRO PRESERVATION AND POST-TRANSFUSION SURVIVAL OF STORED BLOOD. PMID- 16695463 TI - THE MEASUREMENT OF POST-TRANSFUSION SURVIVAL OF PRESERVED STORED HUMAN ERYTHROCYTES BY MEANS OF TWO ISOTOPES OF RADIO-ACTIVE IRON. PMID- 16695464 TI - THE POST-TRANSFUSION SURVIVAL OF PRESERVED HUMAN ERYTHROCYTES STORED AS, WHOLE BLOOD OR IN RESUSPENSION, AFTER REMOVAL OF PLASMA, BY MEANS OF TWO ISOTOPES OF RADIOACTIVE IRON. PMID- 16695465 TI - THE RATE OF POST-TRANSFUSION LOSS OF NON-VIABLE STORED HUMAN ERYTHROCYTES AND THE RE-UTILIZATION OF HEMOGLOBIN-DERIVED RADIO-ACTIVE IRON. PMID- 16695466 TI - THE EFFECT OF VARYING TEMPERATURES ON THE POST-TRANSFUSION SURVIVAL OF WHOLE BLOOD DURING DEPOT STORAGE AND AFTER TRANSPORTATION BY LAND AND AIR. PMID- 16695467 TI - ANTITULARENSE SERUM: CORRELATION BETWEEN PROTECTIVE CAPACITY FOR WHITE RATS AND PRECIPITABLE ANTIBODY CONTENT. PMID- 16695468 TI - THE ROLE OF PEPSIN, PEPTIC INHIBITORY SUBSTANCES, AND HYDROCHLORIC ACID IN NORMAL SUBJECTS AND IN THE PRODUCTION OF PEPTIC ULCERS. PMID- 16695469 TI - THE TOLERANCE FOR POTASSIUM IN SEVERE RENAL IN-SUFFICIENCY: A STUDY OF TEN CASES. PMID- 16695470 TI - THE EFFECTS OF VAGOTOMY ON THE PHYSIOLOGY OF THE STOMACH IN PATIENTS WITH PEPTIC ULCER. PMID- 16695471 TI - METABOLIC ALTERATIONS FOLLOWING THERMAL BURNS. VII. EFFECT OF FORCE-FEEDING, METHIONINE, AND TESTOSTERONE PROPIONATE ON NITROGEN BALANCE IN EXPERIMENTAL BURNS. PMID- 16695472 TI - THE BILIARY EXCRETION OF VARIOUS BILE ACIDS BY PATIENTS WITH CHOLEDOCHOSTOMY DRAINAGE, AND THE EFFECT OF ORAL ADMINISTRATION OF DESICCATED BILE OF THE OX AND THE HOG. PMID- 16695473 TI - THE EFFECT OF GLYCINE ON THE PRODUCTION AND EXCRETION OF URIC ACID. PMID- 16695474 TI - ELECTROPHORETIC ANALYSIS OF ANTE- AND POSTMORTEM SERUM IN DIFFERENT DISEASES. PMID- 16695475 TI - THE IMPORTANCE OF VOLUME AND OF TONICITY OF THE BODY FLUIDS IN SALT DEPLETION SHOCK. PMID- 16695476 TI - THE EFFECT OF INCREASED INTRA-ABDOMINAL PRESSURE ON RENAL FUNCTION IN MAN. PMID- 16695477 TI - THE DIALYZABILITY OF THE PRESSOR AND ANTIDIURETIC ACTIVITIES OF PITRESSIN. PMID- 16695478 TI - PROTEINS IN THE COLLOIDAL GOLD REACTION. PMID- 16695479 TI - THE EFFECT OF LOCAL COOLING ON THE FILTRATION AND ABSORPTION OF FLUID IN THE HUMAN FOREARM. PMID- 16695480 TI - ABNORMAL RENAL TUBULAR BACK-DIFFUSION FOLLOWING ANURIA. PMID- 16695481 TI - THE EFFECT OF CYTOCHROME C UPON THE RESPIRATION OF TISSUE SLICES. PMID- 16695482 TI - THE PLASMA NON-PROTEIN NITROGEN DISTRIBUTION AND ITS CORRELATION WITH THE EFFICACY OF FLUID REPLACEMENT THERAPY FOLLOWING THERMAL INJURY. PMID- 16695483 TI - THE INFLUENCE OF DENITROGENATION ON THE RESPONSE OF ANESTHETIZED DOGS TO INTRAVENOUSLY INJECTED OXYGEN. PMID- 16695484 TI - THE CONJUGATED, NON-PROTEIN, AMINO ACIDS OF PLASMA. IV. A DIFFERENCE IN THE UTILIZATION OF THE PEPTIDES OF HYDROLYSATES OF FIBRIN AND CASEIN. PMID- 16695485 TI - THE CONJUGATED, NON-PROTEIN, AMINO ACIDS OF PLASMA. V. A STUDY OF THE CLINICAL SIGNIFICANCE OF PEPTIDEMIA. PMID- 16695486 TI - STUDIES OF BLOOD VOLUME IN THE TETRALOGY OF FALLOT AND IN OTHER TYPES OF CONGENITAL HEART DISEASE. PMID- 16695487 TI - THE RELATION BETWEEN THE SERUM UREA CONCENTRATION AND THE PROTEIN CONSUMPTION OF NORMAL INDIVIDUALS. PMID- 16695488 TI - THE SERUM CREATININE CONCENTRATION OF NORMAL INDIVIDUALS. PMID- 16695489 TI - A CLINICAL METHOD FOR THE APPROXIMATE DETERMINATION OF SERUM CREATININE CONCENTRATION. PMID- 16695490 TI - THE CORRELATION BETWEEN THE ALBUMIN AND ALPHA GLOBULIN CONTENTS OF PLASMA. PMID- 16695491 TI - BIOCHEMICAL AND HEMODYNAMIC CHANGES FOLLOWING THE SUBCUTANEOUS INJECTION OF GLUCOSE SOLUTION. PMID- 16695492 TI - ON THE CONCENTRATION OF PROTEIN IN SAMPLES OF NORMAL URINE MEASURED BY ITS SURFACE ACTIVITY. PMID- 16695493 TI - THE EFFECT OF CYTOCHROME C UPON THE METABOLISM OF RAT TISSUES. PMID- 16695494 TI - THE RENAL CLEARANCE OF PENICILLINS F, G, K, AND X IN RABBITS AND MAN. PMID- 16695495 TI - THE BLOOD LEVELS AND RENAL CLEARANCE IN RABBITS AND MAN OF AN ANTIBIOTIC DERIVED FROM B. SUBTILIS (BACITRACIN). PMID- 16695496 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XXXV. THE PERSISTENCE OF FIBRIN FILM IN TISSUE AND ITS MODIFICATION BY HEAT TREATMENT. PMID- 16695497 TI - STUDIES ON EXPERIMENTAL PHOSGENE POISONING. III. OXYGEN THERAPY IN PHOSGENE POISONED DOGS AND RATS. PMID- 16695498 TI - THE EFFECTS OF CONTINUOUS PRESSURE BREATHING ON KIDNEY FUNCTION. PMID- 16695499 TI - LIVER BLOOD FLOW IN PREGNANCY-HEPATIC VEIN CATHETERIZATION. PMID- 16695502 TI - THE ESTIMATION OF SERUM SODIUM FROM BICARBONATE PLUS CHLORIDE. PMID- 16695503 TI - THE TRANSPORT AND EXCRETION OF URIC ACID IN MAN. I. TRUE URIC ACID IN NORMAL CEREBROSPINAL FLUID, IN PLASMA, AND IN ULTRAFILTRATES OF PLASMA. PMID- 16695504 TI - THE TRANSPORT AND EXCRETION OF URIC ACID IN MAN. II. THE ENDOGENOUS URIC ACID LIKE CHROMOGEN OF BIOLOGICAL FLUIDS. PMID- 16695505 TI - THE EFFECT OF FATAL P. KNOWLESI MALARIA ON SIMIAN CIRCULATORY AND BODY FLUID COMPARTMENT PHYSIOLOGY. PMID- 16695506 TI - THE OCCURRENCE OF JAUNDICE IN THERAPEUTIC AND NATURAL MALARIA. PMID- 16695507 TI - MECHANISM AND SIGNIFICANCE OF THE THYMOL TURBIDITY TEST FOR LIVER DISEASE. PMID- 16695508 TI - THE EFFECTS OF TEMPORARY CESSATION OF RENAL BLOOD FLOW IN RATS. PMID- 16695509 TI - SEROLOGICAL RESPONSE TO INTRANASAL ADMINISTRATION OF INACTIVE INFLUENZA VIRUS IN CHILDREN. PMID- 16695510 TI - THE VOLUME OF DISTRIBUTION OF MANNITOL AS A MEASURE OF THE VOLUME OF EXTRACELLULAR FLUID, WITH A STUDY OF THE MANNITOL METHOD. PMID- 16695511 TI - EFFECTS OF EXPERIMENTAL HYPERTONIA ON CIRCULATING LEUKOCYTES. PMID- 16695512 TI - THE RELATIONSHIP OF SODIUM CHLORIDE TO HYPERTENSION. PMID- 16695513 TI - CAPILLARY PERMEABILITY IN RELATION TO ACUTE ANOXIA AND TO VENOUS OXYGEN SATURATION. PMID- 16695514 TI - THE HEPATIC BLOOD FLOW AND SPLANCHNIC OXYGEN CONSUMPTION OF MAN-THEIR ESTIMATION FROM UREA PRODUCTION OR BROMSULPHALEIN EXCRETION DURING CATHETERIZATION OF THE HEPATIC VEINS. PMID- 16695515 TI - THE URINARY EXCRETION OF RADIOIODINE IN VARIOUS THYROID STATES. PMID- 16695516 TI - STUDIES ON PAIN: DISCRIMINATION OF DIFFERENCES IN INTENSITY OF A PAIN STIMULUS AS A BASIS OF A SCALE OF PAIN INTENSITY. PMID- 16695517 TI - THE EFFECT OF URINE VOLUME ON UREA EXCRETION. PMID- 16695518 TI - THE INFLUENCE OF ORALLY ADMINISTERED ALKALI AND ACID ON THE RENAL EXCRETION OF QUINACRINE, CHLOROQUINE AND SANTOQUINE. PMID- 16695519 TI - RENAL PLASMA FLOW AND SODIUM REABSORPTION AND EXCRETION IN CONGESTIVE HEART FAILURE. PMID- 16695520 TI - EFFECT OF EXERCISE ON CARDIAC OUTPUT AND PULMONARY ARTERIAL PRESSURE IN NORMAL PERSONS AND IN PATIENTS WITH CARDIOVASCULAR DISEASE AND PULMONARY EMPHYSEMA. PMID- 16695521 TI - LIVER INVOLVEMENT IN INFECTIOUS MONONUCLEOSIS. PMID- 16695522 TI - STUDY OF THE DISAPPEARANCE OF CONGO RED FROM THE BLOOD OF NON-AMYLOID SUBJECTS AND PATIENTS WITH AMYLOIDOSIS. PMID- 16695523 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XXXIV. COMPARATIVE STUDIES ON THE NUTRITIVE VALUE OF ORALLY AND INTRAVENOUSLY ADMINISTERED HUMAN SERUM ALBUMIN IN MAN. PMID- 16695524 TI - THE EFFECTS OF INTRAVENOUS INJECTION OF CONCENTRATED HUMAN SERUM ALBUMIN UPON BLOOD PLASMA, ASCITES AND RENAL FUNCTIONS IN THREE PATIENTS WITH CIRRHOSIS OF THE LIVER. PMID- 16695525 TI - OSMOTIC FACTORS INFLUENCING THE FORMATION OF ASCITES IN PATIENTS WITH CIRRHOSIS OF THE LIVER. PMID- 16695526 TI - VARIATIONS IN THE BLOOD PRESSURE RESPONSE TO REPEATED ADMINISTRATION OF TETRAETHYL AMMONIUM CHLORIDE. PMID- 16695527 TI - THE NATURE OF THE COLD PRESSOR TEST AND ITS SIGNIFICANCE IN RELATION TO NEUROGENIC AND HUMORAL MECHANISMS IN HYPERTENSION. PMID- 16695528 TI - A STUDY OF THE HUMAN MYOGRAM. A STUDY OF NORMALS, AND OF PATIENTS WITH ADDISON'S DISEASE, THYROTOXICOSIS AND PROGRESSIVE MUSCULAR ATROPHY. PMID- 16695529 TI - THE EFFECT OF TETRAETHYLAMMONIUM ON THE SMALL BOWEL OF MAN. PMID- 16695530 TI - THE PROTHROMBIN RESPONSE TO THE PARENTERAL ADMINISTRATION OF LARGE DOSES OF VITAMIN K IN SUBJECTS WITH NORMAL LIVER FUNCTION AND IN CASES OF LIVER DISEASE: A STANDARDIZED TEST FOR THE ESTIMATION OF HEPATIC FUNCTION. PMID- 16695531 TI - THE RENAL REGULATION OF ACID-BASE BALANCE IN MAN. I. THE NATURE OF THE MECHANISM FOR ACIDIFYING THE URINE. PMID- 16695532 TI - THE RENAL REGULATION OF ACID-BASE BALANCE IN MAN. II. FACTORS AFFECTING THE EXCRETION OF TITRATABLE ACID BY THE NORMAL HUMAN SUBJECT. PMID- 16695533 TI - EXCHANGES OF SODIUM AND POTASSIUM IN FAMILIAL PERIODIC PARALYSIS. PMID- 16695534 TI - TRANSFERS OF CELL SODIUM AND POTASSIUM IN EXPERIMENTAL AND CLINICAL CONDITIONS. PMID- 16695535 TI - STUDIES IN SERUM ELECTROLYTES. XV. THE CALCIUM-BINDING PROPERTY OF THE SERUM PROTEINS (MULTIPLE MYELOMA, LYMPHOGRANULOMA VENEREUM AND SARCOIDOSIS). PMID- 16695536 TI - SERUM PRECIPITABLE IODINE CONCENTRATIONS DURING PREGNANCY. PMID- 16695537 TI - STUDIES OF HEMOPHILIA. II. THE ASSAY OF THE ANTIHEMOPHILIC CLOT-PROMOTING PRINCIPLE IN NORMAL HUMAN PLASMA WITH SOME OBSERVATIONS ON THE RELATIVE POTENCY OF CERTAIN PLASMA FRACTIONS. PMID- 16695538 TI - THE ORAL AND PARENTERAL PHENYLALANINE REQUIREMENTS FOR NITROGEN EQUILIBRIUM IN MAN. PMID- 16695539 TI - DEPRESSION OF THE EXOGENOUS CREATININE/INULIN OR THIOSULFATE CLEARANCE RATIOS IN MAN BY DIODRAST AND p-AMINOHIPPURIC ACID. PMID- 16695540 TI - ON THE BLOOD LACTIC ACID RESPONSE TO MEASURED EXERCISE IN HYPOXIC HUMAN SUBJECTS. PMID- 16695541 TI - FACTORS AFFECTING THE APPEARANCE AND PERSISTENCE OF VISIBLE CUTANEOUS REACTIVE HYPEREMIA IN MAN. PMID- 16695543 TI - THE INFLUENCE OF CLOTHING, WORK, AND AIR MOVEMENT ON THE THERMAL EXCHANGES OF ACCLIMATIZED MEN IN VARIOUS HOT ENVIRONMENTS. PMID- 16695542 TI - THE RELATION OF SERUM BICARBONATE CONCENTRATION TO MUSCLE COMPOSITION. PMID- 16695544 TI - FURTHER STUDIES OF THE EFFECTS OF INSULIN ON THE METABOLISM OF VITAMIN C. PMID- 16695545 TI - THE ABSORPTION OF ORALLY ADMINISTERED EMULSIFIED LIPID IN NORMAL CHILDREN AND IN CHILDREN WITH STEATORRHEA. PMID- 16695546 TI - CLINICAL APPLICATION OF A SIMPLE METHOD FOR ESTIMATING "GAMMA GLOBULIN". PMID- 16695547 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XXXVI. INACTIVATION OF THE VIRUS OF HOMOLOGOUS SERUM HEPATITIS IN SOLUTIONS OF NORMAL HUMAN SERUM ALBUMIN BY MEANS OF HEAT. PMID- 16695548 TI - THE ANEMIA OF INFECTION. VII. THE SIGNIFICANCE OF FREE ERYTHROCYTE PROTOPORPHYRIN, TOGETHER WITH SOME OBSERVATIONS ON THE MEANING OF THE "EASILY SPLIT-OFF" IRON. PMID- 16695549 TI - THE SERUM CHOLESTEROL LEVEL OF THE PREMATURELY BORN INFANT AND ITS MOTHER. PMID- 16695550 TI - DEPHOSPHORYLATION OF ADENOSINETRIPHOSPHATE BY NORMAL AND PATHOLOGICAL HUMAN SERA. PMID- 16695551 TI - THE EFFECT OF EXERCISE ON THE RENAL PLASMA FLOW AND FILTRATION RATE OF NORMAL AND CARDIAC SUBJECTS. PMID- 16695552 TI - SENSITIVITY OF THE TUBERCLE BACILLUS TO STREPTOMYCIN BEFORE AND DURING SPECIFIC THERAPY. PMID- 16695553 TI - PLASMA VOLUME, TOTAL CIRCULATING PROTEIN, AND "AVAILABLE FLUID" ABNORMALITIES IN PREECLAMPSIA AND ECLAMPSIA. PMID- 16695554 TI - CARDIOVASCULAR REACTIONS TO EMOTIONAL STIMULI. EFFECT ON THE CARDIAC OUTPUT, ARTERIOVENOUS OXYGEN DIFFERENCE, ARTERIAL PRESSURE, AND PERIPHERAL RESISTANCE. PMID- 16695555 TI - MINERAL BALANCE DURING BRIEF STARVATION. THE EFFECT ON SERUM ELECTROLYTES AND MINERAL BALANCE OF MAINTAINING THE INTAKE OF CERTAIN MINERAL CONSTITUENTS. PMID- 16695556 TI - THE RELATION OF ALBUMIN TO PRECIPITABLE IODINE OF SERUM. PMID- 16695557 TI - NITROGEN BALANCE STUDIES ON THE KEMPNER RICE DIET. PMID- 16695558 TI - THE EFFECTS OF THE RATE OF ADMINISTRATION OF AMINO ACID PREPARATIONS ON URINARY WASTAGE OF AMINO ACID NITROGEN IN MAN. PMID- 16695559 TI - THE COMPARATIVE EFFECTS OF CONTINUOUS AND INTERMITTENT PENICILLIN THERAPY ON THE FORMATION OF ANTISTREPTOLYSIN IN HEMOLYTIC STREPTOCOCCAL PHARYNGITIS. PMID- 16695560 TI - QUANTITATIVE ANTISTREPTOKINASE STUDIES IN PATIENTS INFECTED WITH GROUP A HEMOLYTIC STREPTOCOCCI: A COMPARISON WITH SERUM ANTISTREPTOLYSIN AND GAMMA GLOBULIN LEVELS WITH SPECIAL REFERENCE TO THE OCCURRENCE OF RHEUMATIC FEVER. PMID- 16695561 TI - THE EFFECT OF HEPARIN AND DICUMAROL ANTICOAGULANT THERAPY UPON THE ERYTHROCYTE SEDIMENTATION RATE. PMID- 16695562 TI - SENSITIVITY OF SKELETAL MUSCLE TO INTRA-ARTERIAL ACETYLCHOLINE IN NORMAL AND MYASTHENIC MAN. PMID- 16695563 TI - RESPONSE OF CITRIC ACID LEVELS TO ORAL ADMINISTRATION OF GLUCOSE. I. NORMAL ADULTS AND CHILDREN. PMID- 16695564 TI - RESPONSE OF CITRIC ACID LEVELS TO ORAL ADMINISTRATION OF GLUCOSE. II. ABNORMALITIES OBSERVED IN THE DIABETIC AND CONVULSIVE STATE. PMID- 16695565 TI - THE EFFECTS OF VARIOUS AMINO ACIDS ON PERIPHERAL BLOOD FLOW AND SKIN TEMPERATURE. PMID- 16695566 TI - STUDIES OF THE MUCIN-CLOT PREVENTION TEST FOR THE DETERMINATION OF THE ANTIHYALURONIDASE TITRE OF HUMAN SERUM. PMID- 16695567 TI - ANTIHYALURONIDASE STUDIES OF SERA FROM PATIENTS WITH RHEUMATIC FEVER, STREPTOCOCCAL INFECTIONS, AND MISCELLANEOUS NON-STREPTOCOCCAL DISEASES. PMID- 16695568 TI - THE NITROUS OXIDE METHOD FOR THE QUANTITATIVE DETERMINATION OF CEREBRAL BLOOD FLOW IN MAN: THEORY, PROCEDURE AND NORMAL VALUES. PMID- 16695570 TI - THE EFFECTS OF INCREASED INTRACRANIAL PRESSURE ON CEREBRAL CIRCULATORY FUNCTIONS IN MAN. PMID- 16695569 TI - THE EFFECTS OF ALTERED ARTERIAL TENSIONS OF CARBON DIOXIDE AND OXYGEN ON CEREBRAL BLOOD FLOW AND CEREBRAL OXYGEN CONSUMPTION OF NORMAL YOUNG MEN. PMID- 16695571 TI - THE BLOOD FLOW AND OXYGEN CONSUMPTION OF THE HUMAN BRAIN IN DIABETIC ACIDOSIS AND COMA. PMID- 16695572 TI - THE BLOOD FLOW, VASCULAR RESISTANCE, AND OXYGEN CONSUMPTION OF THE BRAIN IN ESSENTIAL HYPERTENSION. PMID- 16695573 TI - THE URINARY EXCRETION OF INSULIN BY NORMAL AND DIABETIC SUBJECTS. PMID- 16695574 TI - THE EFFECT OF ANEMIA AND POLYCYTHEMIA ON DIGITAL INTRAVASCULAR BLOOD VISCOSITY. PMID- 16695575 TI - HOMOLOGOUS AND HETEROLOGOUS ANTIBODY RESPONSE OF INFANTS AND CHILDREN TO MULTIPLE INJECTIONS OF A SINGLE STRAIN OF INFLUENZA VIRUS. PMID- 16695576 TI - THE EFFECT OF SPONTANEOUS AND ARTIFICIALLY INDUCED FEVER ON LIVER FUNCTION. PMID- 16695577 TI - THE EFFECTS OF THE CARDIAC GLYCOSIDES UPON THE DYNAMICS OF THE CIRCULATION IN CONGESTIVE HEART FAILURE. I. OUABAIN. PMID- 16695578 TI - THE BILIARY EXCRETION OF BROMSULFALEIN AS A TEST OF LIVER FUNCTION IN A GROUP OF PATIENTS FOLLOWING HEPATITIS OR SERUM JAUNDICE. PMID- 16695579 TI - STUDIES ON THE MUCOPROTEINS OF HUMAN PLASMA. I. DETERMINATION AND ISOLATION. PMID- 16695582 TI - THE EVALUATION OF AN EFFECTIVE DOSAGE OF CARONAMIDE (4 CARBOXYPHENYLMETHANESULFONANILIDE) FOR THE SUPPRESSION OF TUBULAR EXCRETION OF PENICILLIN IN CHILDREN. PMID- 16695581 TI - AN ESTIMATION OF THE HEPATIC BLOOD FLOW AND SPLANCHNIC OXYGEN CONSUMPTION IN HEART FAILURE. PMID- 16695583 TI - RENAL OXYGEN CONSUMPTION IN MAN DURING ABDOMINAL COMPRESSION. PMID- 16695584 TI - THE EFFECT OF EXERCISE ON RENAL PLASMA FLOW IN NORMAL MALE SUBJECTS. PMID- 16695585 TI - THE RENAL CLEARANCE OF ENDOGENOUS "CREATININE" IN MAN. PMID- 16695586 TI - STUDIES ON AMINO ACID METABOLISM. II. BLOOD GLYCINE AND TOTAL AMINO ACIDS IN VARIOUS PATHOLOGICAL CONDITIONS, WITH OBSERVATIONS ON THE EFFECTS OF INTRAVENOUSLY ADMINISTERED GLYCINE. PMID- 16695587 TI - STUDIES ON AMINO ACID METABOLISM. III. PLASMA GLYCINE CONCENTRATION AND HIPPURIC ACID FORMATION FOLLOWING THE INGESTION OF BENZOATE. PMID- 16695588 TI - THE AURICULOTEMPORAL SYNDROME. A CLINICAL AND PHARMACOLOGIC STUDY. PMID- 16695589 TI - STUDIES IN METHIONINE AND SULFUR METABOLISM. I. THE FATE OF INTRAVENOUSLY ADMINISTERED METHIONINE, IN NORMAL INDIVIDUALS AND IN PATIENTS WITH LIVER DAMAGE. PMID- 16695590 TI - MEASUREMENT OF GLOMERULAR FILTRATION RATE IN PREMATURE INFANTS. PMID- 16695591 TI - ESTIMATIONS OF THE DECREASE IN EFFECTIVE BLOOD VOLUME WHEN PRESSURE BREATHING AT SEA LEVEL. PMID- 16695592 TI - RESISTANCE TO THE ACTION OF THE ENDOTOXINS OF ENTERIC BACILLI IN MAN. PMID- 16695593 TI - COMPARISON OF THE CONSTANT INFUSION AND URINE COLLECTION TECHNIQUES FOR THE MEASUREMENT OF RENAL FUNCTION. PMID- 16695594 TI - EVALUATION OF NEUROGENIC AND HUMORAL FACTORS IN BLOOD PRESSURE MAINTENANCE IN NORMAL AND TOXEMIC PREGNANCY USING TETRAETHYLAMMONIUM CHLORIDE. PMID- 16695595 TI - URINARY EXCRETION OF AMINO ACIDS FOLLOWING THE RAPID INJECTION OF A SOLUTION OF AMINO ACIDS IN MAN. PMID- 16695597 TI - THE INTRAVENOUS GLUCOSE TOLERANCE TEST IN PREGNANCY. PMID- 16695596 TI - A METHOD FOR THE QUANTITATIVE DETERMINATION OF THE CEPHALIN-CHOLESTEROL FLOCCULATION REACTION. PMID- 16695598 TI - THE GENETICS OF GOUT AND HYPERURICEMIA-AN ANALYSIS OF NINETEEN FAMILIES. PMID- 16695600 TI - THE EFFECT OF SODIUM CHLORIDE DEPLETION ON BLOOD PRESSURE AND TETRAETHYLAMMONIUM CHLORIDE RESPONSE IN HYPERTENSION. PMID- 16695599 TI - STUDIES ON CARBOHYDRATE METABOLISM IN PATIENTS WITH GASTRIC CANCER. DEFECTIVE HEPATIC GLYCOGENESIS; EFFECTS OF ADRENO-CORTICAL EXTRACT. PMID- 16695601 TI - THROMBIN FORMATION. I. THE ROLE OF CALCIUM, SERUM AC-GLOBULIN AND TISSUE THROMBOPLASTIN. PMID- 16695602 TI - A STUDY OF ANTIFIBRINOLYSIN ACTIVITY IN THE PLASMAS OF VARIOUS ANIMAL SPECIES. PMID- 16695603 TI - A STUDY OF THE ANTIFIBRINOLYSIN ACTIVITY IN HUMAN PLASMAS DURING PATHOLOGICAL STATES. PMID- 16695604 TI - BLOOD VOLUME DETERMINATION IN THE HUMAN WITH RED CELLS CONTAINING RADIOACTIVE PHOSPHORUS (P) AND WITH PURE HUMAN ALBUMIN. PMID- 16695605 TI - THE SPECIFICITY OF IMMUNE HUMAN SERUM ANTIHYALURONIDASE. PMID- 16695606 TI - RENAL AND CIRCULATORY FACTORS IN THE EDEMA FORMATION OF CONGESTIVE HEART FAILURE. PMID- 16695607 TI - UROPEPSIN EXCRETION BY MAN. I. THE SOURCE, PROPERTIES AND ASSAY OF UROPEPSIN. PMID- 16695608 TI - UROPEPSIN EXCRETION BY MAN. II. UROPEPSIN EXCRETION BY HEALTHY MEN. PMID- 16695609 TI - UROPEPSIN EXCRETION BY MAN. III. UROPEPSIN EXCRETION BY PATIENTS WITH PEPTIC ULCER AND OTHER LESIONS OF THE STOMACH. PMID- 16695610 TI - EFFECTS OF HYPOXIA AND HYPERCAPNIA ON PERCEPTION OF THERMAL CUTANEOUS PAIN. PMID- 16695611 TI - THE USE OF CONCENTRATED HUMAN SERUM ALBUMIN IN THE TREATMENT OF CIRRHOSIS OF THE LIVER. PMID- 16695612 TI - THE COMPLEMENT CONTENT OF HUMAN SERA WITH ESPECIAL REFERENCE TO MALARIA. PMID- 16695613 TI - LUNG FUNCTION STUDIES. I. THE RATE OF INCREASE OF ARTERIAL OXYGEN SATURATION DURING THE INHALATION OF 100 PER CENT O(2). PMID- 16695614 TI - THE EFFECT OF SPINAL ANESTHESIA ON THE RENAL ISCHEMIA IN CONGESTIVE HEART FAILURE. PMID- 16695616 TI - CHANGES IN CEREBROSPINAL FLUID PRESSURE UNDER THE INFLUENCE OF CONTINUOUS SUBARACHNOIDAL INFUSION OF NORMAL SALINE. PMID- 16695615 TI - THE CHANGES IN THE SERUM PROTEINS IN PATIENTS WITH EXPERIMENTALLY INDUCED INFECTIOUS HEPATITIS. PMID- 16695617 TI - BLOOD AND EXTRACELLULAR FLUID STUDIES IN CHRONIC MALNUTRITION IN INFANCY. PMID- 16695618 TI - STUDIES ON GANGRENE FOLLOWING COLD INJURY. IX. THE EFFECT OF RUTIN AND OTHER CHEMICAL AGENTS ON THE COURSE OF EXPERIMENTAL FROSTBITE IN RABBITS. PMID- 16695619 TI - KIDNEY FUNCTION IN ADRENAL INSUFFICIENCY. PMID- 16695620 TI - STUDIES ON PAIN: AN INVESTIGATION OF SOME QUANTITATIVE ASPECTS OF THE DOL SCALE OF PAIN INTENSITY. PMID- 16695621 TI - STUDIES ON THE CHEMOTHERAPY OF THE HUMAN MALARIAS. VII. THE ANTIMALARIAL ACTIVITY OF PAMAQUINE. PMID- 16695622 TI - STUDIES ON THE CHEMOTHERAPY OF THE HUMAN MALARIAS. VIII. THE PHYSIOLOGICAL DISPOSITION OF PAMAQUINE. PMID- 16695623 TI - THE USE OF SN-10,275 IN THE PROPHYLAXIS AND TREATMENT OF SPOROZOITE-INDUCED VIV AX MALARIA (CHESSON STRAIN). PMID- 16695624 TI - STUDIES ON THE CHEMOTHERAPY OF THE HUMAN MALARIAS. IX. EFFECT OF PAMAQUINE ON THE BLOOD CELLS OF MAN. PMID- 16695625 TI - STUDIES ON THE CHEMOTHERAPY OF THE HUMAN MALARIAS. X. THE SUPPRESSIVE ANTIMALARIAL EFFECT OF PALUDRINE. PMID- 16695626 TI - A TECHNIQUE FOR THE DETECTION OF MINIMAL NUMBERS OF MALARIA PARASITES; ITS APPLICATION IN THE DETECTIN OF SUPPRESSED VIVAX MALARIA. PMID- 16695627 TI - METHEMALBUMIN. I. APPEARANCE DURING ADMINISTRATION OF PAMAQUINE AND QUININE. PMID- 16695628 TI - METHEMALBUMIN. II. EFFECT OF PAMAQUINE AND QUININE ON PATHWAYS OF HEMOGLOBIN METABOLISM. PMID- 16695629 TI - THE TOXICITY OF LARGE DOSES OF PENTAQUINE (SN-13,276), A NEW ANTIMALARIAL DRUG. PMID- 16695630 TI - PROCEDURES USED AT STATEVILLE PENITENTIARY FOR THE TESTING OF POTENTIAL ANTIMALARIAL AGENTS. PMID- 16695631 TI - PENTAQUINE (SN-13,276), A THERAPEUTIC AGENT EFFECTIVE IN REDUCING THE RELAPSE RATE IN VIV AX MALARIA. PMID- 16695632 TI - THE CLINICAL TRIAL OF EIGHTEEN ANALOGUES OF PAMAQUIN (PLASMOCHIN) IN VIV AX MALARIA (CHESSON STRAIN). PMID- 16695633 TI - COMPARISON OF CHLOROQUINE, QUINACRINE (ATABRINE), AND QUININE IN THE TREATMENT OF ACUTE ATTACKS OF SPOROZOITE-INDUCED VIV AX MALARIA (CHESSON STRAIN). PMID- 16695634 TI - THE THERAPEUTIC EFFECTIVENESS OF LARGE DOSES OF PALUDRINE IN ACUTE ATTACKS OF SPOROZOITE-INDUCED VIV AX MALARIA (CHESSON STRAIN). PMID- 16695635 TI - A LICHEN-PLANUS-LIKE ERUPTION OCCURRING DURING THE COURSE OF CHLOROQUINE ADMINISTRATION. PMID- 16695636 TI - A STUDY OF THE PROPHYLACTIC EFFECTIVENESS OF SEVERAL 8-AMINOQUINOLINES IN SPOROZOITE-INDUCED VIVAX MALARIA (CHESSON STRAIN). PMID- 16695637 TI - STUDIES ON THE CHRONIC TOXICITY OF CHLOROQUINE (SN-7618). PMID- 16695638 TI - STUDIES ON THE CHEMOTHERAPY OF THE HUMAN MALARIAS. I. METHOD FOR THE QUANTITATIVE ASSAY OF SUPPRESSIVE ANTIMALARIAL ACTION IN VIVAX MALARIA. PMID- 16695639 TI - STUDIES ON THE CHEMOTHERAPY OF THE HUMAN MALARIAS. II. METHOD FOR THE QUANTITATIVE ASSAY OF SUPPRESSIVE ANTIMALARIAL ACTION IN FALCIPARUM MALARIA. PMID- 16695640 TI - STUDIES ON THE CHEMOTHERAPY OF THE HUMAN MALARIAS. III. THE PHYSIOLOGICAL DISPOSITION AND ANTIMALARIAL ACTIVITY OF THE CINCHONA ALKALOIDS. PMID- 16695641 TI - STUDIES ON THE CHEMOTHERAPY OF THE HUMAN MALARIAS. IV. THE METABOLISM OF CINCHONINE IN RELATION TO ITS ANTIMALARIAL ACTIVITY. PMID- 16695642 TI - STUDIES ON THE CHEMOTHERAPY OF THE HUMAN MALARIAS. V. THE ANTIMALARIAL ACTIVITY OF QUINACRINE. PMID- 16695643 TI - STUDIES ON THE CHEMOTHERAPY OF THE HUMAN MALARIAS. VI. THE PHYSIOLOGICAL DISPOSITION, ANTIMALARIAL ACTIVITY, AND TOXICITY OF SEVERAL DERIVATIVES OF 4 AMINOQUINOLINE. PMID- 16695644 TI - STUDIES IN DIABETIC ACIDOSIS AND COMA, WITH PARTICULAR EMPHASIS ON THE RETENTION OF ADMINISTERED POTASSIUM. PMID- 16695645 TI - SIMULTANEOUS MEASUREMENT OF EFFECTIVE RENAL BLOOD FLOW AND CARDIAC OUTPUT IN RESTING NORMAL SUBJECTS AND PATIENTS WITH ESSENTIAL HYPERTENSION. PMID- 16695646 TI - CAN VASCULAR SHUNTING BE INDUCED IN THE KIDNEY BY VASOACTIVE DRUGS? PMID- 16695647 TI - ELECTROPHRENIC RESPIRATION. II. ITS USE IN MAN. PMID- 16695648 TI - THE VOLUME OF THE LUNG DETERMINED BY HELIUM DILUTION. DESCRIPTION OF THE METHOD AND COMPARISON WITH OTHER PROCEDURES. PMID- 16695649 TI - STUDIES ON HUMAN ALCAPTONURIA. EFFECT OF THIOURACIL, PARA-AMINOBENZOIC ACID AND DI-IODOTYROSINE ON EXCRETION OF HOMOGENTISIC ACID. PMID- 16695650 TI - THE SIGNIFICANCE OF THE DIFFERENCE IN SYSTEMIC ARTERIAL AND VENOUS PLASMA CONCENTRATIONS IN RENAL CLEARANCE METHODS. PMID- 16695651 TI - STUDIES ON PAIN: MEASUREMENTS OF PAIN INTENSITY IN CHILDBIRTH. PMID- 16695652 TI - METHODS FOR MEASURING THE ACTIVITY OF COMPONENTS OF THE STREPTOCOCCAL FIBRINOLYTIC SYSTEM, AND STREPTOCOCCAL DESOXYRIBONUCLEASE. PMID- 16695653 TI - THE EFFECT IN PATIENTS OF STREPTOCOCCAL FIBRINOLYSIN (STREPTOKINASE) AND STREPTOCOCCAL DESOXYRIBONUCLEASE ON FIBRINOUS, PURULENT, AND SANGUINOUS PLEURAL EXUDATIONS. PMID- 16695654 TI - USE OF BLOOD DONORS WITH POSITIVE SEROLOGIC TESTS FOR SYPHILIS-WITH A NOTE ON THE DISAPPEARANCE OF PASSIVELY TRANSFERRED REAGIN. PMID- 16695655 TI - HUMAN PROTHROMBIN: QUANTITATIVE STUDIES ON THE PLASMA LABILE FACTOR AND THE RESTORATIVE EFFECTS OF NORMAL, HYPOFI-BRINOGENEMIC, AND HEMOPHILIC PLASMA ON THE PROTHROMBIN OF STORED PLASMA. PMID- 16695656 TI - A STUDY OF THE RATE OF DISAPPEARANCE OF A DIGITALIS GLYCOSIDE (LANATOSIDE C.) FROM THE BLOOD OF MAN. PMID- 16695657 TI - THE RENAL REGULATION OF ACID-BASE BALANCE IN MAN. III. THE REABSORPTION AND EXCRETION OF BICARBONATE. PMID- 16695658 TI - THE BIOASSAY OF ADRENAL CORTICOIDS IN THE URINE OF PATIENTS WITH CONGESTIVE HEART FAILURE. PMID- 16695659 TI - AUTOANTIBODIES IN HUMAN GLOMERULONEPHRITIS. PMID- 16695660 TI - THE HYALURONIC ACID OF SYNOVIAL FLUID IN RHEUMATOID ARTHRITIS. PMID- 16695661 TI - BLOOD OXYGEN STUDIES IN PATIENTS WITH POLYCYTHEMIA AND IN NORMAL SUBJECTS. PMID- 16695662 TI - THE PHOSPHORUS TURNOVER OF CARCINOMA OF THE HUMAN STOMACH AS MEASURED WITH RADIOACTIVE PHOSPHORUS. PMID- 16695663 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XXXVII. THE METAL-COMBINING GLOBULIN OF HUMAN PLASMA. PMID- 16695664 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XXXVIII. SERUM IRON TRANSPORT. MEASUREMENT OF IRON-BINDING CAPACITY OF SERUM IN MAN. PMID- 16695665 TI - CHEMICAL, CLINICAL, AND IMMUNOLOGICAL STUDIES ON THE PRODUCTS OF HUMAN PLASMA FRACTIONATION. XXXIX. THE ANEMIA OF INFECTION. STUDIES ON THE IRON-BINDING CAPACITY OF SERUM. PMID- 16695666 TI - POTASSIUM DEFICIENCY AND THE ROLE OF THE KIDNEY IN ITS PRODUCTION. PMID- 16695667 TI - THE BEHAVIOR OF RADIOIODINE IN THE BLOOD. PMID- 16695668 TI - INDUCED INSULIN RESISTANCE IN THE RABBIT. PMID- 16695669 TI - RELATIVE MEASUREMENT IN VIVO OF ACCUMULATION OF RADIOIODINE BY THE HUMAN THYROID GLAND: COMPARISON WITH RADIOACTIVITY IN PERIPHERAL TISSUES. PMID- 16695670 TI - THE MEASUREMENT OF THE IODINE-ACCUMULATING FUNCTION OF THE HUMAN THYROID GLAND. PMID- 16695671 TI - THE VALIDITY OF THE ELECTROKYMOGRAPHIC METHOD FOR MEASUREMENT OF DIAMETER CHANGE OF THE AORTA AND PULMONARY ARTERY DURING CIRCULATORY DISTURBANCE. PMID- 16695672 TI - STUDIES OF THE ROLE OF THE LIVER IN HUMAN CARBOHYDRATE METABOLISM BY THE VENOUS CATHETER TECHNIC. I. NORMAL SUBJECTS UNDER FASTING CONDITIONS AND FOLLOWING THE INJECTION OF GLUCOSE. PMID- 16695673 TI - METABOLIC STUDIES ON PROTEIN-DEPLETED PATIENTS RECEIVING A LARGE PART OF THEIR NITROGEN INTAKE FROM HUMAN SERUM ALBUMIN ADMINISTERED INTRAVENOUSLY. PMID- 16695674 TI - CONGENITAL METHEMOGLOBINEMIA. A CLINICAL AND BIOCHEMICAL STUDY OF A CASE. PMID- 16695676 TI - COMPLEMENT AND ITS COMPONENTS IN HUMAN CEREBROSPINAL FLUID. PMID- 16695677 TI - HEMATOLOGIC OBSERVATIONS IN PATIENTS WITH CHRONIC HEPATIC INSUFFICIENCY: STERNAL BONE MARROW MORPHOLOGY AND BONE MARROW PLASMACYTOSIS. PMID- 16695678 TI - THE CARDIOVASCULAR EFFECTS OF ACUTELY INDUCED HYPOTHERMIA. PMID- 16695679 TI - THE CIRCULATORY RESPONSES TO HYPERTHERMIA INDUCED BY RADIANT HEAT. PMID- 16695680 TI - THE RELATION OF INFLUENZA VIRUS AND BACTERIA IN THE ETIOLOGY OF PNEUMONIA. PMID- 16695681 TI - DETERMINATION OF THE PLASMA GLYCINE AFTER GELATIN FEEDING AS A DIAGNOSTIC PROCEDURE FOR PANCREATIC FIBROSIS. PMID- 16695682 TI - STUDIES IN CIRRHOSIS OF THE LIVER. I. RELATIONSHIP BETWEEN PLASMA VOLUME, PLASMA PROTEIN CONCENTRATIONS AND TOTAL CIRCULATING PROTEINS. PMID- 16695683 TI - THE AGGLUTINATION OF AUTOCLAVED HEMOLYTIC STREPTOCOCCI BY SERUM FROM PATIENTS WITH RHEUMATIC FEVER AND OTHER CONDITIONS. PMID- 16695684 TI - THE CLOTTING BEHAVIOR OF HUMAN "PLATELET-FREE" PLASMA: EVIDENCE FOR THE EXISTENCE OF A "PLASMA THROMBOPLASTIN". PMID- 16695685 TI - THE HEMODYNAMIC EFFECTS OF HYPOTENSIVE DRUGS IN MAN. I. VERATRUM VIRIDE. PMID- 16695686 TI - SOME OBSERVATIONS ON THE BALLISTOCARDIOGRAPHIC PATTERN, WITH SPECIAL REFERENCE TO THE H AND K WAVES. PMID- 16695687 TI - TRANSFERS OF POTASSIUM IN RENAL INSUFFICIENCY. PMID- 16695688 TI - COMPLEMENT AND ITS COMPONENT FRACTIONS IN CEREBROSPINAL FLUID IN INFLAMMATORY CEREBROSPINAL DISEASES. PMID- 16695689 TI - THE EFFECT OF ADRENOCORTICOTROPHIC HORMONE IN CONGENITAL ADRENAL HYPERPLASIA WITH VIRILISM AND IN CUSHING'S SYNDROME TREATED WITH METHYL TESTOSTERONE. PMID- 16695690 TI - A BRIEF SKETCH OF THE HISTORY OF THE AMERICAN SOCIETY FOR CLINICAL INVESTIGATION. PMID- 16695691 TI - SOME OBSERVATIONS ON THE DEVELOPMENT OF HYPOKALIEMIA DURING THERAPY OF DIABETIC ACIDOSIS IN JUVENILE AND YOUNG ADULT SUBJECTS. PMID- 16695692 TI - THE EFFECT OF BILATERAL STELLATE GANGLION BLOCK ON THE CEREBRAL CIRCULATION IN NORMOTENSIVE AND HYPERTENSIVE PATIENTS. PMID- 16695693 TI - PEPTIDASE ACTIVITY IN HUMAN SERUM FOLLOWING BONE FRACTURE. PMID- 16695694 TI - THE RENAL REGULATION OF ACID-BASE BALANCE IN MAN. IV. THE NATURE OF THE RENAL COMPENSATIONS IN AMMONIUM CHLORIDE ACIDOSIS. PMID- 16695695 TI - THE EFFECTS OF HISTAMINE ON RENAL FUNCTION IN HYPERTENSIVE AND NORMOTENSIVE SUBJECTS. PMID- 16695696 TI - STUDIES OF COPROPORPHYRIN. I. THE PER DIEM EXCRETION AND ISOMER DISTRIBUTION OF COPROPORPHYRIN IN NORMAL HUMAN URINE. PMID- 16695697 TI - STUDIES OF COPROPORPHYRIN. II. AN INVESTIGATION OF THE CONTRIBUTION TO THE URINARY COPROPORPHYRIN OF HEMOGLOBIN AND OF BACTERIAL METABOLISM IN THE INTESTINAL TRACT. PMID- 16695698 TI - STUDIES OF COPROPORPHYRIN. III. IDIOPATHIC COPROPORPHYRINURIA; A HITHERTO UNRECOGNIZED FORM CHARACTERIZED BY LACK OF SYMPTOMS IN SPITE OF THE EXCRETION OF LARGE AMOUNTS OF COPROPORPHYRIN. PMID- 16695700 TI - III. THE FATE OF CITRATE IN ERYTHROBLASTOTIC INFANTS TREATED WITH EXCHANGE TRANSFUSION. PMID- 16695699 TI - INTRA-ARTICULAR TEMPERATURE AS A MEASURE OF JOINT REACTION. PMID- 16695701 TI - ELECTROLYTE CHANGES IN HUMAN STRIATED MUSCLE IN ACIDOSIS AND ALKALOSIS. PMID- 16695702 TI - STUDIES ON FREE ERYTHROCYTE PROTOPORPHYRIN, SERUM IRON, SERUM IRON-BINDING CAPACITY AND PLASMA COPPER DURING NORMAL PREGNANCY. PMID- 16695703 TI - STUDIES ON THE ACTIVATION OF A SERUM "PROTHROMBIN-CONVERTING FACTOR". PMID- 16695704 TI - STUDIES IN IRON TRANSPORTATION AND METABOLISM. VII. EVIDENCE OBTAINED WITH RADIOACTIVE IRON THAT "EASILY SPLIT-OFF" BLOOD IRON IS AN ARTEFACT. PMID- 16695705 TI - A COMPARISON OF THE DIRECT FICK AND THE GROLLMAN METHODS FOR DETERMINATION OF THE CARDIAC OUTPUT IN MAN. PMID- 16695706 TI - A FACTOR DETERMINING THE LOCATION OF PSEUDOFRACTURES IN OSTEOMALACIA. PMID- 16695707 TI - THE OXYGEN CONSUMPTION OF THE NORMAL AND THE DISEASED HUMAN KIDNEY. PMID- 16695708 TI - THE MEASUREMENT OF GLOMERULAR AND TUBULAR PLASMA FLOW IN THE NORMAL AND DISEASED HUMAN KIDNEY. PMID- 16695709 TI - HYPERVOLEMIC ANEMIA IN CIRRHOSIS. PMID- 16695710 TI - THE EFFECT OF HYALURONIDASE ON THE ABSORPTION OF PARENTERALLY ADMINISTERED RADIOACTIVE PLASMA PROTEINS IN THE DOG. PMID- 16695712 TI - THE EFFECT OF EXERCISE AND BODY POSITION ON THE VENOUS PRESSURE AT THE ANKLE IN PATIENTS HAVING VENOUS VALVULAR DEFECTS. PMID- 16695711 TI - THE ACCELERATION OF LINEAR FLOW IN THE DEEP VEINS OF THE LOWER EXTREMITY OF MAN BY LOCAL COMPRESSION. PMID- 16695713 TI - THE INHIBITION OF FROSTBITE WHEALS BY THE IONTOPHORESIS OF ANTIHISTAMINIC AGENTS. PMID- 16695714 TI - THE CONSTRUCTION OF NORMAL STANDARDS FOR CARDIAC OUTPUT IN MAN. PMID- 16695715 TI - AN EVALUATION OF HUMAN SERUM ALBUMIN IN THE TREATMENT OF CIRRHOSIS OF THE LIVER. PMID- 16695716 TI - THE EFFECT OF HUMAN SERUM ALBUMIN, MERCURIAL DIURETICS, AND A LOW SODIUM DIET ON SODIUM EXCRETION IN PATIENTS WITH CIRRHOSIS OF THE LIVER. PMID- 16695717 TI - IMPROVEMENT OF ACTIVE LIVER CIRRHOSIS IN PATIENTS MAINTAINED WITH AMINO ACIDS INTRAVENOUSLY AS THE SOURCE OF PROTEIN AND LIPOTROPIC SUBSTANCES. PMID- 16695718 TI - THE EFFECTS OF DIHYDROERGOCORNINE ON THE CIRCULATION IN THE EXTREMITIES OF MAN. PMID- 16695719 TI - STUDIES OF COPROPORPHYRIN. IV. THE PER DIEM EXCRETION AND ISOMER DISTRIBUTION IN THE URINE IN INFECTIOUS HEPATITIS, INFECTIOUS MONONUCLEOSIS, AND MECHANICAL JAUNDICE. PMID- 16695720 TI - STUDIES ON VASOMOTOR TONE. I. THE EFFECT OF THE TETRA-ETHYLAMMONIUM ION ON THE PERIPHERAL BLOOD FLOW OF NORMAL SUBJECTS. PMID- 16695721 TI - THE EFFECT OF ANOXIC ANOXIA ON THE HUMAN KIDNEY. PMID- 16695722 TI - DETERMINATION OF ADRENERGIC AND CHOLINERGIC SUBSTANCES IN THE BLOOD OF MAN. PMID- 16695723 TI - CONCENTRATION-TIME COURSE IN THE PLASMA OF MAN OF RADIOMERCURY INTRODUCED AS A MERCURIAL DIURETIC. PMID- 16695724 TI - A STUDY OF CERTAIN ASPECTS OF BLOOD COAGULATION IN THE POSTOPERATIVE STATE IN CONGESTIVE HEART FAILURE AND IN THROMBOPHLEBITIS. PMID- 16695725 TI - STUDIES OF LUNG VOLUMES AND INTRAPULMONARY MIXING: NOTES ON OPEN-CIRCUIT METHODS, INCLUDING USE OF A NEW PIVOTED TYPE GASOMETER FOR LUNG CLEARANCE STUDIES. PMID- 16695726 TI - OBSERVATIONS ON THE PROTEOLYTIC ACTIVITY IN VITRO AT NEUTRAL REACTION OF GASTRIC JUICE FROM PATIENTS WITH SPRUE. PMID- 16695728 TI - TREATMENT OF NEPHROSIS WITH CONCENTRATED HUMAN SERUM ALBUMIN. I. EFFECTS ON THE PROTEINS OF BODY FLUIDS. PMID- 16695727 TI - SODIUM AND CHLORIDE DEPLETION IN ACUTE PORPHYRIA WITH REFERENCE TO THE STATUS OF ADRENAL CORTICAL FUNCTION. PMID- 16695729 TI - MUCOLYTIC ENZYME SYSTEMS. VIII. INHIBITION OF HYALURONIDASE BY HUMAN BLOOD SERUM DURING THE NORMAL MENSTRUAL CYCLE AND PREGNANCY. PMID- 16695730 TI - STUDIES ON AMINO ACID EXCRETION IN MAN. III. AMINO ACID LEVELS IN PLASMA AND URINE OF NORMAL MEN FED DIETS OF VARYING PROTEIN CONTENT. PMID- 16695731 TI - FACTORS IN THE TREATMENT OF LAENNEC'S CIRRHOSIS. I. CLINICAL AND HISTOLOGICAL CHANGES OBSERVED DURING A CONTROL PERIOD OF BED-REST, ALCOHOL WITH-DRAWAL, AND A MINIMAL BASIC DIET. PMID- 16695732 TI - OXYGEN SATURATION OF STERNAL MARROW BLOOD IN POLYCYTHEMIA VERA. PMID- 16695733 TI - IV. RESPONSE OF CITRIC ACID LEVELS OF NORMAL ADULTS AND CHILDREN TO INTRAMUSCULAR INJECTION OF EPINEPHRINE. PMID- 16695734 TI - STUDIES OF THE EFFECTS OF FLAVONOIDS ON ROENTGEN IRRADIATION DISEASE. II. COMPARISON OF THE PROTECTIVE INFLUENCE OF SOME FLAVONOIDS AND VITAMIN C IN DOGS. PMID- 16695735 TI - THE PLASMA VISCOSITY IN PULMONARY TUBERCULOSIS AND RHEUMATIC DISEASES. PMID- 16695736 TI - FURTHER EXPERIENCES WITH INTRAVENOUS AUREOMYCIN THERAPY. A STUDY OF 116 CASES. PMID- 16695737 TI - PURITY STUDIES ON POLYPEPTIDE ANTIBIOTICS: BACITRACIN. PMID- 16695738 TI - A COMPARATIVE STUDY OF THE RENAL DAMAGE PRODUCED IN MICE BY VARIOUS LOTS OF BACITRACIN. PMID- 16695739 TI - CHEMICAL STUDIES ON POLYMYXIN B. PMID- 16695740 TI - LABORATORY AND CLINICAL STUDIES OF POLYMYXIN B AND E. PMID- 16695741 TI - THE PHARMACOLOGY OF CIRCULIN. PMID- 16695742 TI - PREPARATION OF A MODIFIED SUBTILIN SUITABLE FOR THE TREATMENT OF TUBERCULOSIS AND OTHER INFECTIONS IN ANIMALS. PMID- 16695743 TI - THE ELIMINATION OF PENICILLIN G IN BILATERALLY NEPHRECTOMIZED DOGS. PMID- 16695744 TI - STUDIES ON HYPERSENSITIVITY TO PENICILLIN. I. INCIDENCE OF REACTIONS IN 1303 PATIENTS. PMID- 16695745 TI - THE RECOVERY OF BACTERIA FROM THE TOXIC EFFECTS OF PENICILLIN. PMID- 16695746 TI - AMPEROMETRIC MICRO-TITRATION OF STREPTOMYCIN AND DIHYDROSTREPTOMYCIN. PMID- 16695747 TI - A NEW LINEAR DIFFUSION METHOD FOR THE MICROBIOLOGICAL ASSAY OF STREPTOMYCIN AND DIHYDROSTREPTOMYCIN. PMID- 16695748 TI - QUANTITATIVE DETERMINATION OF DIHYDROSTREPTOMYCIN BY PERIODATE OXIDATION. PMID- 16695749 TI - INVESTIGATIONS ON THE STREPTOMYCINS. PMID- 16695750 TI - THE EFFECT OF STREPTOMYCIN DERIVATIVES ON STREPTOMYCIN-DEPENDENT AND -RESISTANT STRAINS OF BACTERIA. PMID- 16695751 TI - THE DEVELOPMENT OF RESISTANCE OF SALMONELLA TYPHOSA TO THE FATTY ACID SALTS OF STREPTOMYCIN AND DIHYDROSTREPTOMYCIN. PMID- 16695752 TI - THE EFFECTS OF NORMAL HORSE SERUM ON THE IN VITRO ACTIVITY OF TYROTHRICIN. PMID- 16695753 TI - IN VITRO STUDIES ON POSSIBLE SYNERGISTIC ACTION BETWEEN PENICILLIN AND BACITRACIN. PMID- 16695754 TI - COMPARISON OF THE ACTION OF STREPTOMYCIN, POLYMYXIN B, AUREOMYCIN AND CHLOROMYCETIN ON H. PERTUSSIS, H. PARAPERTUSSIS, H. INFLUENZAE AND FIVE ENTERIC STRAINS OF GRAM-NEGATIVE BACILLI. PMID- 16695755 TI - EXPERIMENTAL STUDIES ON THE ACTION OF STREPTOMYCIN, AUREOMYCIN, AND CHLOROMYCETIN ON BRUCELLA. PMID- 16695756 TI - ANTIGENIC AND ENZYME SYSTEM CHANGES IN BETA HEMOLYTIC STREPTOCOCCI RESISTANT TO PENICILLIN, STREPTOMYCIN, BACITRACIN AND AUREOMYCIN. PMID- 16695757 TI - PATTERNS OF BACTERIAL RESISTANCE TO PENICILLIN, AUREOMYCIN, AND STREPTOMYCIN. PMID- 16695758 TI - THE PARTIAL PURIFICATION AND PROPERTIES OF ANTIBIOTIC SUBSTANCES FROM THE SWEET POTATO PLANT (IPOMOEA BATATAS). PMID- 16695759 TI - THE PARTIAL PURIFICATION AND PROPERTIES OF ANTIBIOTIC SUBSTANCES FROM THE BANANA (MUSA SAPIENTUM). PMID- 16695760 TI - ANTIBIOTIC STUDIES ON AN EXTRACT FROM LEPTOTAENIA MULTIFEDA. PMID- 16695762 TI - LUPULON AND HUMULON-ANTIBIOTIC CONSTITUENTS OF HOPS. PMID- 16695761 TI - FACTORS INFLUENCING THE ANTIBIOTIC ACTIVITY OF LUPULON. PMID- 16695763 TI - THE OCCURRENCE OF ANTIBACTERIAL SUBSTANCES ACTIVE AGAINST MYCOBACTERIUM TUBERCULOSIS IN SEED PLANTS. PMID- 16695764 TI - A CRYSTALLINE ANTIFUNGAL AGENT, MYCOSUBTILIN, ISOLATED FROM SUBTILIN BROTH. PMID- 16695765 TI - THE ANTIFUNGAL ACTIVITY OF ANTIBIOTIC XG. PMID- 16695766 TI - NEOMYCIN-PRODUCTION AND ANTIBIOTIC PROPERTIES. PMID- 16695767 TI - DIFFUSION PLATE ASSAY FOR CHLORAMPHENICOL AND AUREOMYCIN. PMID- 16695768 TI - CHLORAMPHENICOL (CHLOROMYCETIN), AN ANTIBIOTIC. PHARMACOLOGICAL AND PATHOLOGICAL STUDIES IN ANIMALS. PMID- 16695769 TI - SUSCEPTIBILITY OF MICRO-ORGANISMS TO CHLORAMPHENICOL (CHLOROMYCETIN). PMID- 16695770 TI - PRELIMINARY REPORT ON THE USE OF CHLORAMPHENICOL (CHLOROMYCETIN) IN THE TREATMENT OF ACUTE GONORRHEAL URETHRITIS. PMID- 16695771 TI - THE BENEFICIAL EFFECT OF CHLOROMYCETIN IN BRUCELLOSIS. PMID- 16695772 TI - THE EFFECT OF AUREOMYCIN AGAINST BACTERIUM TULARENSE. PMID- 16695773 TI - THE CONCENTRATION OF AUREOMYCIN IN URINE AND CEREBROSPINAL, PLEURAL AND ASCITIC FLUIDS AFTER ORAL AND INTRAVENOUS ADMINISTRATION. PMID- 16695774 TI - CLINICAL EXPERIENCE WITH AUREOMYCIN. PMID- 16695775 TI - THE CLINICAL EVALUATION OF AUREOMYCIN. PMID- 16695777 TI - EVIDENCE THAT RENAL SODIUM EXCRETION BY NORMAL HUMAN SUBJECTS IS REGULATED BY ADRENAL CORTICAL ACTIVITY. PMID- 16695776 TI - THE SERUM PROTEINS IN INFECTIOUS MONONUCLEOSIS. ELECTROPHORETIC STUDIES. PMID- 16695778 TI - SOME EFFECTS OF VARIATION IN SODIUM INTAKE AND OF DIFFERENT SODIUM SALTS IN NORMAL SUBJECTS. PMID- 16695780 TI - THE ACTION OF STREPTOCOCCAL DESOXYRIBOSE NUCLEASE (STREPTODORNASE)IN VITRO AND ON PURULENT PLEURAL EXUDATIONS OF PATIENTS. PMID- 16695779 TI - FURTHER STUDIES WITH THE SALT TOLERANCE TEST IN NORMAL INDIVIDUALS AND IN PATIENTS WITH ADRENAL CORTICAL HYPERFUNCTION. PMID- 16695781 TI - STUDIES WITH RADIOACTIVE IODIZED FAT. I. PREPARATION OF RADIOACTIVE FAT WITH OBSERVATIONS ON THE ABSORPTION OF FAT FOLLOWING SUBCUTANEOUS AND INTRAPERITONEAL INJECTION IN DOGS. PMID- 16695782 TI - STUDIES WITH RADIOACTIVE IODIZED FAT. II. THE TISSUE DISTRIBUTION OF EMULSIFIED FAT FOLLOWING INTRAVENOUS ADMINISTRATION. PMID- 16695783 TI - THE MEASUREMENT OF EXTRACELLULAR FLUID IN MAN BY MEANS OF A CONSTANT INFUSION TECHNIQUE. PMID- 16695784 TI - STUDIES OF THE ROLE OF THE LIVER IN HUMAN CARBOHYDRATE METABOLISM BY THE VENOUS CATHETER TECHNIC. II. PATIENTS WITH DIABETIC KETOSIS, BEFORE AND AFTER THE ADMINISTRATION OF INSULIN. PMID- 16695785 TI - STUDIES OF THE PULMONARY CIRCULATION. I. THE CIRCULATION TIME FROM THE PULMONARY ARTERY TO THE FEMORAL ARTERY AND THE QUANTITY OF BLOOD IN THE LUNGS IN NORMAL INDIVIDUALS. PMID- 16695786 TI - STUDIES OF THE PULMONARY CIRCULATION. II. THE CIRCULATION TIME FROM THE PULMONARY ARTERY TO THE FEMORAL ARTERY AND THE QUANTITY OF BLOOD IN THE LUNGS IN PATIENTS WITH MITRAL STENOSIS AND IN PATIENTS WITH LEFT VENTRICULAR FAILURE. PMID- 16695787 TI - MATURATION OF RENAL FUNCTION IN CHILDHOOD: CLEARANCE STUDIES. PMID- 16695788 TI - THE CEREBRAL BLOOD FLOW IN MALE SUBJECTS AS MEASURED BY THE NITROUS OXIDE TECHNIQUE. NORMAL VALUES FOR BLOOD FLOW, OXYGEN UTILIZATION, GLUCOSE UTILIZATION, AND PERIPHERAL RESISTANCE, WITH OBSERVATIONS ON THE EFFECT OF TILTING AND ANXIETY. PMID- 16695789 TI - SEROLOGICAL STUDIES IN RHEUMATIC FEVER. II. SERUM COMPLEMENT IN THE RHEUMATIC STATE. PMID- 16695790 TI - EVALUATION OF THERAPEUTIC SUBSTANCES EMPLOYED FOR THE RELIEF OF BRONCHOSPASM. V. ADRENERGIC AGENTS. PMID- 16695791 TI - EVALUATION OF THERAPEUTIC SUBSTANCES EMPLOYED FOR THE RELIEF OF BRONCHOSPASM. VI. AMINOPHYLLINE. PMID- 16695792 TI - CHLORAMPHENICOL (CHLOROMYCETIN) IN THE TREATMENT OF TSUTSUGAMUSHI DISEASE (SCRUB TYPHUS). PMID- 16695793 TI - CHANGES IN LIVER GLYCOGEN STUDIED BY THE NEEDLE ASPIRATION TECHNIC IN PATIENTS WITH DIABETIC KETOSIS. WITH A METHOD FOR THE ESTIMATION OF GLYCOGEN FROM HISTOLOGIC PREPARATIONS. PMID- 16695794 TI - COMPARISONS OF THE DISTRIBUTION OF RADIOACTIVE IODINE IN SERUM AND URINE IN DIFFERENT LEVELS OF THYROID FUNCTION. PMID- 16695795 TI - EFFECTS OF ADRENOCORTICOTROPHIC HORMONE ON NEURO-MUSCULAR FUNCTION IN PATIENTS WITH MYASTHENIA GRAVIS. PMID- 16695796 TI - THE SERUM PROTEINS IN PORTAL CIRRHOSIS UNDER MEDICAL MANAGEMENT. ELECTROPHORETIC STUDIES. PMID- 16695797 TI - INVESTIGATION OF THE AMINOACIDURIA IN WILSON'S DISEASE (HEPATOLENTICULAR DEGENERATION): DEMONSTRATION OF A DEFECT IN RENAL FUNCTION. PMID- 16695798 TI - OBSERVATIONS ON THE CHANGE OF VENTRICULAR SYSTOLE (QT INTERVAL) DURING EXERCISE. PMID- 16695799 TI - COMPARATIVE STUDY OF THE EFFECTS OF TETRAETHYL-AMMONIUM CHLORIDE AND VERATRUM VIRIDE ON BLOOD PRESSURE IN NORMAL AND TOXEMIC PREGNANCY. PMID- 16695800 TI - THE INSULIN TOLERANCE TEST IN PATIENTS WITH ESSENTIAL HYPERTENSION. PMID- 16695801 TI - URINARY EXCRETION OF AMINO ACIDS IN LIVER DISEASE. PMID- 16695802 TI - A RAPID METHOD FOR DETERMINING BLOOD VOLUMES BY THE USE OF P-LABELLED RED CELLS. PMID- 16695803 TI - ACTIVATION OF PROPLASMIN BY A FACTOR FROM MAMMALIAN TISSUE. PMID- 16695804 TI - THE METABOLISM OF l-TYROSINE IN INFANTILE SCURVY. PMID- 16695805 TI - TRANSFUSION OF LEUKOCYTES LABELED WITH RADIOACTIVE PHOSPHORUS. PMID- 16695806 TI - RENAL VENOUS PRESSURE IN CHRONIC CONGESTIVE HEART FAILURE. PMID- 16695807 TI - EXCRETION OF 11-OXYCORTICOSTEROIDS IN PARAPLEGIC AND RHEUMATOID ARTHRITIC PATIENTS. PMID- 16695808 TI - SEROLOGIC RESPONSE TO STREPTOCOCCAL HEMOLYSIN AND HYALURONIDASE IN STREPTOCOCCAL AND RHEUMATIC INFECTION. PMID- 16695809 TI - SERUM CITRIC ACID AFTER INGESTION OF GLUCOSE IN NORMAL AND STARVED SUBJECTS. PMID- 16695810 TI - STUDIES ON DIURETICS. I. THE SITE OF ACTION OF MERCURIAL DIURETICS. PMID- 16695811 TI - STUDIES ON DIURETICS. II. THE RELATIONSHIP BETWEEN GLOMERULAR FILTRATION RATE, PROXIMAL TUBULAR ABSORPTION OF SODIUM AND DIURETIC EFFICACY OF MERCURIALS. PMID- 16695812 TI - COMPARISON OF THE VOLUME OF DISTRIBUTION, RENAL AND EXTRARENAL CLEARANCES OF INULIN AND MANNITOL IN MAN. PMID- 16695813 TI - THE COMPARATIVE EFFECTS OF SMALL INTRAVENOUS DOSES OF L-NOR-EPINEPHRINE UPON ARTERIAL PRESSURE AND PULSE RATE IN NOR-MOTENSIVE SUBJECTS AND IN HYPERTENSIVE PATIENTS BEFORE AND AFTER THORACOLUMBAR SYMPATHECTOMY. PMID- 16695815 TI - The Renal Lesion in Epidemic Hemorrhagic Fever. PMID- 16695814 TI - LUNG FUNCTION STUDIES. IV. POSTURAL CHANGES IN RESPIRATORY DEAD SPACE AND FUNCTIONAL RESIDUAL CAPACITY. PMID- 16695816 TI - The Renal Lesion in Epidemic Hemorrhagic Fever. PMID- 16695817 TI - INTRODUCTION. PMID- 16695818 TI - CONSTITUTION OF THE AMERICAN SOCIETY FOR CLINICAL INVESTIGATION (AS AMENDED TO 1959). PMID- 16695820 TI - Roster of the American Society for Clinical Investigation 1959. PMID- 16695819 TI - OFFICERS OF THE SOCIETY FROM ITS ORGANIZATION TO THE PRESENT TIME. PMID- 16695821 TI - AMERICAN SOCIETY FOR CLINICAL INVESTIGATION. ROSTER OF EDITORIAL STAFFS OF THE JOURNAL OF CLINICAL INVESTIGATION FROM 1924 TO THE PRESENT TIME. PMID- 16695822 TI - CORTISOL METABOLISM IN MAN: OBSERVATIONS OF PATHWAYS, POOL SIZES OF METABOLITES AND RATES OF FORMATION OF METABOLITES. PMID- 16695823 TI - DETERMINANTS OF THE FLOW AND COMPOSITION OF BILE IN THE UNANESTHETIZED DOG DURING CONSTANT INFUSIONS OF SODIUM TAUROCHOLATE. PMID- 16695824 TI - A STUDY OF THE RELATIONSHIP OF THE NORMAL BACTERICIDAL ACTIVITY OF HUMAN SERUM TO BACTERIAL INFECTION. PMID- 16695825 TI - Water and Electrolyte Studies in Cholera. PMID- 16695826 TI - IMMUNOLOGICAL STUDIES WITH PLASMA EXPANDERS DERIVED FROM HUMAN PLASMA. PMID- 16695827 TI - CATECHOL AMINE EXCRETION BY DUODENAL ULCER PATIENTS. PMID- 16695828 TI - DELIVERY OF ANTIMICROBIAL DRUGS ACROSS INFLAMMATORY MEMBRANE IN RABBITS. PMID- 16695829 TI - MEASUREMENT OF BLOOD FLOW IN THE COLLATERAL VESSELS OF THE PORTAL VEIN; PRELIMINARY RESULTS OF A NEW METHOD. PMID- 16695830 TI - A LECITHINASE A IN DUODENAL CONTENTS OF MAN. PMID- 16695831 TI - QUANTITATIVE HISTOCHEMISTRY OF THE NEPHRON. III. LACTIC DEHYDROGENASE ACTIVITY IN MAN AND OTHER SPECIES. PMID- 16695832 TI - LEUKOKINETIC STUDIES. II. A METHOD FOR LABELING GRANULOCYTES IN VITRO WITH RADIOACTIVE DIISOPROPYLFLUOROPHOSPHATE (DFP). PMID- 16695833 TI - MEASUREMENT OF SMALL QUANTITIES OF INSULIN-LIKE ACTIVITY USING RAT ADIPOSE TISSUE. II. EVALUATION OF PERFORMANCE. PMID- 16695834 TI - THE CONCENTRATIONS OF CYSTEINE AND CYSTINE IN HUMAN BLOOD PLASMA. PMID- 16695835 TI - X-RAY DIFFRACTION STUDIES OF HUMAN AORTIC ELASTIN RESIDUES. PMID- 16695836 TI - STUDIES OF UNBOUND AMINO ACID DISTRIBUTIONS IN PLASMA, ERYTHROCYTES, LEUKOCYTES AND URINE OF NORMAL HUMAN SUBJECTS. PMID- 16695837 TI - UNBOUND AMINO ACID CONCENTRATIONS IN PLASMA, ERYTHROCYTES, LEUKOCYTES AND URINE OF PATIENTS WITH LEUKEMIA. PMID- 16695838 TI - OBSERVATIONS ON THE FUNCTIONAL HOMOGENEITY OF THE NEPHRON POPULATION IN THE CHRONICALLY DISEASED KIDNEY OF THE DOG. PMID- 16695839 TI - SERUM beta-LIPOPROTEIN LEVELS IN HEALTH AND DISEASE, AS DETERMINED BY THE NEPHELOMETRIC METHOD WITH AMYLOPECTIN SULFATE AS REAGENT. PMID- 16695840 TI - STUDIES ON STARLING'S LAW OF THE HEART. III. OBSERVATIONS IN PATIENTS WITH MITRAL STENOSIS AND ATRIAL FIBRILLATION ON THE RELATIONSHIPS BETWEEN LEFT VENTRICULAR END-DIASTOLIC SEGMENT LENGTH, FILLING PRESSURE, AND THE CHARACTERISTICS OF VENTRICULAR CONTRACTION. PMID- 16695841 TI - THE EFFECT OF CUSHING'S SYNDROME UPON SERUM ALBUMIN METABOLISM. PMID- 16695842 TI - A STUDY OF EIGHT HEATED PLASMA PROTEIN PREPARATIONS. PMID- 16695843 TI - EFFECT OF CHRONIC HYPOTHALAMIC STIMULATION UPON CHOLESTEROL-INDUCED ATHEROSCLEROSIS IN THE RABBIT. PMID- 16695844 TI - THE EFFECT OF ACETYLCHOLINE UPON RESPIRATORY GAS EXCHANGE IN MITRAL STENOSIS. PMID- 16695845 TI - A LABILE SERUM FACTOR CLOTTING DEFECT: ITS DEMONSTRATION BY THE THROMBOPLASTIN GENERATION TEST AND ITS CLINICAL SIGNIFICANCE. PMID- 16695846 TI - STUDIES ON DIGITALIS. IV. OBSERVATIONS IN MAN ON THE EFFECTS OF DIGITALIS PREPARATIONS ON THE CONTRACTILITY OF THE NON-FAILING HEART AND ON TOTAL VASCULAR RESISTANCE. PMID- 16695847 TI - QUANTITATIVE HISTOCHEMISTRY OF THE NEPHRON. VI. HYDROXYPROLINE IN THE HUMAN GLOMERULUS. PMID- 16695848 TI - SERUM FACTORS OF ACQUIRED HEMOLYTIC ANEMIA IN LEUKEMIA AND LYMPHOMA. PMID- 16695849 TI - DIRECT RENAL ACTION OF SOME DIGITALIS STEROIDS. PMID- 16695850 TI - THE EFFECTS OF INTRAVENOUS INFUSIONS OF VALINE-5 ANGIOTENSIN II AND OTHER PRESSOR AGENTS ON URINARY ELECTROLYTES AND CORTICOSTEROIDS, INCLUDING ALDOSTERONE. PMID- 16695851 TI - PULMONARY FUNCTION IN HYPERTHYROIDISM. PMID- 16695852 TI - STUDIES OF GLYCOGEN METABOLISM IN LIVER GLYCOGEN DISEASE (VON GIERKE'S DISEASE): SIX CASES WITH SIMILAR METABOLIC ABNORMALITIES AND RESPONSES TO GLUCAGON. PMID- 16695853 TI - STUDIES OF BONE OF PATIENTS WITH HYPERPARATHYDROIDISM BY MEANS OF PHOSPHATE EXCHANGE EXPERIMENTS IN VITRO AND QUANTITATIVE MICRORADIOGRAPHY. PMID- 16695854 TI - Comparative studies on the half-life of I-labeled albumins and nonradioactive human serum albumin in a case of analbuminemia. PMID- 16695855 TI - THE INFLUENCE OF INTRACARDIAC BARORECEPTORS ON VENOUS RETURN, SYSTEMIC VASCULAR VOLUME AND PERIPHERAL RESISTANCE. PMID- 16695856 TI - THE EFFECT OF VITAMIN SUPPLEMENTATION ON THE URINARY EXCRETION OF TRYPTOPHAN METABOLITES BY PREGNANT WOMEN. PMID- 16695857 TI - THE EFFECT OF IN VIVO AGING OF NORMAL HUMAN ERYTHROCYTES AND ERYTHROCYTE MACROMOLECULES UPON OXYHEMOGLOBIN DISSOCIATION. PMID- 16695858 TI - PYRIMIDINE METABOLISM IN MAN. IV. THE ENZYMATIC DEFECT OF OROTIC ACIDURIA. PMID- 16695859 TI - THE EFFECTS OF THERAPY ON PULMONARY MECHANICS IN HUMAN PULMONARY EDEMA. PMID- 16695860 TI - DETECTION OF THE HETEROZYGOUS CARRIER OF THE WILSON'S DISEASE GENE. PMID- 16695862 TI - Proceedeings of the Fifty-Third Annual Meeting of the American Society for Clinical Investigation, Inc., Held in Atlantic City, N. J., May 1, 1961. PMID- 16695861 TI - EFFECTS OF TRIPARANOL (MER-29) ON CHOLESTEROL BIOSYNTHESIS AND ON BLOOD STEROL LEVELS IN MAN. PMID- 16695865 TI - CORRELATION OF STRUCTURE AND FUNCTION IN THE HANDLING OF GLUCOSE BY THE NEPHRONS OF THE CANINE KIDNEY. PMID- 16695864 TI - THE NATURE OF THE ACTION OF INTRAVENOUS ALDOSTERONE: EVIDENCE FOR A ROLE OF THE HORMONE IN URINARY DILUTION. PMID- 16695866 TI - EFFECT OF HIGH CALCIUM INTAKE ON STRONTIUM METABOLISM IN MAN. PMID- 16695867 TI - IMMUNOREACTIONS INVOLVING PLATELETS. V. POST-TRANSFUSION PURPURA DUE TO A COMPLEMENT-FIXING ANTIBODY AGAINST A GENETICALLY CONTROLLED PLATELET ANTIGEN. A PROPOSED MECHANISM FOR THROMBOCYTOPENIA AND ITS RELEVANCE IN "AUTOIMMUNITY". PMID- 16695868 TI - THE METABOLISM OF NORMAL PLASMA PROTEINS AND GAMMA-MYELOMA PROTEIN IN MICE BEARING PLASMA-CELL TUMORS. PMID- 16695869 TI - PHENYLTHIOCARBAMIDE NON-TASTING AMONG CONGENITAL ATHYROTIC CRETINS: FURTHER STUDIES IN AN ATTEMPT TO EXPLAIN THE INCREASED INCIDENCE. PMID- 16695870 TI - THE INCORPORATION OF COPPER INTO CERULOPLASMIN IN VIVO: STUDIES WITH COPPER AND COPPER. PMID- 16695871 TI - SOME OBSERVATIONS ON THE METABOLISM OF D-GALACTOSE IN NORMAL MAN. PMID- 16695872 TI - CLINICAL AND PHYSIOLOGICAL OBSERVATIONS IN A PATIENT WITH AN IDIOPATHIC DECREASE IN THE THYROXINE-BINDING GLOBULIN OF PLASMA. PMID- 16695873 TI - THEORY OF THE USE OF ARTERIOVENOUS CONCENTRATION DIFFERENCES FOR MEASURING METABOLISM IN STEADY AND NON-STEADY STATES. PMID- 16695874 TI - DEMONSTRATION OF CIRCULATING ANTINUCLEAR GLOBULINS IN ULCERATIVE COLITIS. PMID- 16695875 TI - STUDIES OF RESPIRATORY PHYSIOLOGY IN CHILDREN. VI. LUNG DIFFUSING CAPACITY, DIFFUSING CAPACITY OF THE PULMONARY MEMBRANE AND PULMONARY CAPILLARY BLOOD VOLUME IN CONGENITAL HEART DISEASE. PMID- 16695876 TI - TOTAL NONDIALYZABLE SOLIDS (TNDS) IN HUMAN URINE. IX. IMMUNOCHEMICAL STUDIES OF THE R-1 "UROMUCOID" FRACTION. PMID- 16695877 TI - THE RENAL EXCRETION OF URIC ACID IN GOUT. PMID- 16695878 TI - TOTAL NONDIALYZABLE SOLIDS (TNDS) IN HUMAN URINE. XIII. IMMUNOLOGICAL DETECTION OF A COMPONENT PECULIAR TO RENAL CALCULOUS MATRIX AND TO URINE OF CALCULOUS PATIENTS. PMID- 16695879 TI - THE ACCELERATION OF THROMBUS FORMATION BY CERTAIN FATTY ACIDS. PMID- 16695880 TI - ACTH LEVELS IN PLASMA IN PREOPERATIVE AND SURGICALLY STRESSED PATIENTS. PMID- 16695881 TI - STUDIES ON HUMAN PROTEINURIA. II. SOME CHARACTERISTICS OF THE GAMMA GLOBULINS EXCRETED IN NORMAL, EXERCISE, POSTURAL, AND NEPHROTIC PROTEINURIA. PMID- 16695882 TI - PRESSURE-VOLUME CHARACTERISTICS OF THE HUMAN COLON. PMID- 16695883 TI - PULMONARY DIFFUSING CAPACITY DURING REST AND EXERCISE. A STUDY OF NORMAL PERSONS AND PERSONS WITH ATRIAL SEPTAL DEFECT, PREGNANCY, AND PULMONARY DISEASE. PMID- 16695884 TI - SITES OF INITIAL REMOVAL OF CHYLOMICRON TRIGLYCERIDE FATTY ACIDS FROM THE BLOOD. PMID- 16695885 TI - IMPAIRMENT OF URIC ACID EXCRETION IN GOUT. PMID- 16695886 TI - THE EFFECTS OF AUTONOMIC NERVOUS SYSTEM INHIBITION ON THE CIRCULATORY RESPONSE TO MUSCULAR EXERCISE. PMID- 16695887 TI - THE MECHANISM OF ANTIDIURESIS ASSOCIATED WITH THE ADMINISTRATION OF HYDROCHLOROTHIAZIDE TO PATIENTS WITH VASOPRESSIN-RESISTANT DIABETES INSIPIDUS. PMID- 16695888 TI - IN VITRO INTERCONVERSION OF 16-C-ESTRONE AND 16-C-ESTRADIOL-17beta BY ERYTHROCYTES FROM NORMAL SUBJECTS AND FROM SUBJECTS WITH A DEFICIENCY OF RED CELL GLUCOSE-6-PHOSPHATE DEHYDROGENASE ACTIVITY. PMID- 16695889 TI - RHEOLOGY OF HUMAN BLOOD. STUDIES ON THE FLOW OF FRESH VENOUS BLOOD AND OF CITRATED BLOOD THROUGH TUBING OF DIAMETERS BETWEEN 0.09 AND 0.20 CM. PMID- 16695890 TI - SEROLOGIC STUDIES OF PROTEOLYTIC FRAGMENTS OF RABBIT AGGLUTINATING ANTIBODIES. PMID- 16695891 TI - HETEROGENEITY OF MYELOMA PROTEINS. PMID- 16695892 TI - THE DYNAMICS OF PULSUS ALTERNANS: ALTERNATING END-DIASTOLIC FIBER LENGTH AS A CAUSATIVE FACTOR. PMID- 16695893 TI - SOME OBSERVATIONS ON THE INFLUENCE OF A MAGNESIUM-DEFICIENT DIET ON RATS, WITH SPECIAL REFERENCE TO RENAL CONCENTRATING ABILITY. PMID- 16695894 TI - THE EXPERIMENTAL INDUCTION IN MAN OF SENSITIVITY TO LEUCINE HYPOGLYCEMIA. PMID- 16695895 TI - THE DETERMINATION OF SODIUM IN BODY FLUIDS BY THE GLASS ELECTRODE. PMID- 16695896 TI - THE PRESSOR EFFECT OF TYRAMINE IN MAN AND ITS MODIFICATION BY RESERPINE PRETREATMENT. PMID- 16695897 TI - IN VIVO BEHAVIOR OF I-FIBRINOGEN. PMID- 16695898 TI - STUDIES ON TYPE-SPECIFIC STREPTOCOCCAL ANTIBODIES AS INDICATORS OF PREVIOUS STREPTOCOCCAL INFECTIONS IN RHEUMATIC AND NONRHEUMATIC CHILDREN. PMID- 16695899 TI - THE "INDUCTION" OF DIHYDROFOLIC REDUCTASE ACTIVITY IN LEUKOCYTES AND ERYTHROCYTES OF PATIENTS TREATED WITH AMETHOPTERIN. PMID- 16695900 TI - BRISKET DISEASE. III. SPONTANEOUS REMISSION OF PULMONARY HYPERTENSION AND RECOVERY FROM HEART FAILURE. PMID- 16695901 TI - ENDOTOXIN TOLERANCE. I. ITS INDUCTION BY EXPERIMENTAL PYELONEPHRITIS. PMID- 16695902 TI - ENDOTOXIN TOLERANCE. II. ITS OCCURRENCE IN PATIENTS WITH PYELONEPHRITIS. PMID- 16695903 TI - STUDIES ON TASTE THRESHOLDS IN NORMAL MAN AND IN PATIENTS WITH ADRENAL CORTICAL INSUFFICIENCY: THE ROLE OF ADRENAL CORTICAL STEROIDS AND OF SERUM SODIUM CONCENTRATION. PMID- 16695904 TI - CELL PROLIFERATION KINETICS IN THE GASTROINTESTINAL TRACT OF MAN. I. CELL RENEWAL IN COLON AND RECTUM. PMID- 16695905 TI - TWO TYPES OF gamma-MYELOMA PROTEINS, beta(2A)-MYELOMA PROTEINS, gamma(1) MACROGLOBULINS, AND BENCE JONES PROTEINS IDENTIFIED BY TWO GROUPS OF COMMON ANTIGENIC DETERMINANTS. PMID- 16695906 TI - PROCEEDINGS OF THE FIFTY-FIFTH ANNUAL MEETING OF THE AMERICAN SOCIETY FOR CLINICAL INVESTIGATION, INC., HELD IN ATLANTIC CITY, N. J., APRIL 29, 1963. PRESIDENTIAL ADDRESS. PMID- 16695907 TI - STUDIES ON DIGITALIS. IX. EFFECTS OF OUABAIN ON THE NONFAILING HUMAN HEART. PMID- 16695908 TI - ALCOHOL HYPOGLYCEMIA. I. CARBOHYDRATE METABOLISM OF PATIENTS WITH CLINICAL ALCOHOL HYPOGLYCEMIA AND THE EXPERIMENTAL REPRODUCTION OF THE SYNDROME WITH PURE ETHANOL. PMID- 16695909 TI - ANGIOTENSIN II, NOREPINEPHRINE, AND RENAL TRANSPORT OF ELECTROLYTES AND WATER IN NORMAL MAN AND IN CIRRHOSIS WITH ASCITES. PMID- 16695911 TI - PROGRAM OF THE FIFTY-SIXTH ANNUAL MEETING OF THE AMERICAN SOCIETY FOR CLINICAL INVESTIGATION, INC., TO BE HELD IN ATLANTIC CITY, NEW JERSEY, MAY 4, 1964. PMID- 16695910 TI - THE TURNOVER OF HEMOGLOBINS A, F, AND A(2) IN THE PERIPHERAL BLOOD OF THREE PATIENTS WITH THALASSEMIA. PMID- 16695912 TI - The Role of Proaccelerin in Human Blood Coagulation. Evidence that Proaccelerin Is Converted to a Prothrombin-converting Principle by Activated Stuart Factor: With Notes on the Anticoagulant Action of Soybean Trypsin Inhibitor, Protamine Sulfate, and Hexadimethrine Bromide. PMID- 16695913 TI - The mechanism of salt wastage in chronic renal disease. PMID- 16695914 TI - In vitro studies of human ragweed allergy: changes in cellular and humoral activity associated with specific desensitization. PMID- 16695915 TI - Biochemical Findings in Normal and Osteoarthritic Articular Cartilage. II. Chondroitin Sulfate Concentration and Chain Length, Water, and Ash Content. PMID- 16695916 TI - Thyroxine Turnover and Transport in Laennec's Cirrhosis of the Liver. AB - Studies of the metabolism of thyroxine in 14 cases of cirrhosis revealed a variety of deviations from normal. In addition to radiothyroxine turnover studies, determinations were made of the free thyroxine fractions and free thyroxine iodine concentrations in serum (magnesium precipitation method) as well as the maximal binding capacities of thyroxine-binding alpha globulin (TBG) and thyroxine-binding prealbumin (TBPA) by reverse flow paper electrophoresis in a glycine acetate system at pH 8.6.All cases of cirrhosis exhibited diminutions in TBPA capacities but their TBG capacities showed a wide scatter (13.4 to 41.6 mug/100 ml). The free thyroxine fraction was quite variable, with distinct elevations in nine of the 17 sera.The binding proteins appeared to be determinants of the free thyroxine fraction, which in turn, appeared to be a direct determinant of the half-time of turnover. These inferences did not exclude other possible factors including diminished hepatic uptake and metabolism of the hormone in liver disease.Despite considerable alterations in biological half times, free thyroxine values, and binding proteins, it was remarkable that the absolute hormone disposal was normal in all 14 patients with cirrhosis. PMID- 16695917 TI - Studies in the ketosis of fasting. AB - A series of experiments was performed during the induction of starvation ketosis and in the acute reversal of the ketotic state. In contrast to the predictions of two widely held theories of ketogenesis, control of acetoacetate production by the liver appeared to be unrelated to changes in fatty acid mobilization from the periphery, fatty acid oxidation, fatty acid synthesis, or the acetyl coenzyme A concentration in the liver.Ketosis of fasting was shown to be reversible within 5 minutes by the injection of glucose or insulin. This effect was due to a prompt cessation of acetoacetate production by the liver. The possibility is raised that the ketosis of fasting is due to a direct activation of acetoacetate-synthesizing enzymes secondary to a starvation-induced depression of insulin secretion by the pancreas. PMID- 16695918 TI - Plasma volume in cirrhosis of the liver: its relation of portal hypertension, ascites, and renal failure. AB - Plasma volume was measured by using albumin-(131)I- and (51)Cr-labeled erythrocytes in 24 control subjects, 140 patients with hepatic cirrhosis, and 10 patients with various portal-systemic shunts for the relief of noncirrhotic portal hypertension. The cirrhotic patients included subgroups with ascites, functional renal failure, and portacaval anastomoses. Elevated values for plasma volume, by both methods, were found in each group of patients.The lymph space drained by the thoracic duct was measured by a radioisotopic technique in six patients with cirrhosis and ascites. The amount of radioactivity in this space was found to be negligible in accounting for the elevated plasma volume. Similar results were obtained when the degree of leakage of albumin-(131)I into the ascites was determined in 10 patients with cirrhosis.The plasma volume was unusually elevated in patients who had bled from esophageal varices, and paired comparisons before and after portacaval shunt normal values. There was a statistically significant correlation between normal values. There was a statistically significant correlation between plasma volume and wedged hepatic venous pressure measured in 36 patients.We concluded that the elevated values for plasma volume in cirrhosis are valid and are not artifacts due to leakage of albumin-(131)I from the circulation during mixing. We also concluded that portal hypertension is responsible for the plasma volume expansion; however, we were unable on this basis to explain the failure of portacaval shunting to return the plasma volume to normal, unless the shunt or some other factor keeps the plasma volume elevated. PMID- 16695919 TI - Renal bicarbonate reabsorption and hydrogen ion excretion in normal infants. AB - After acute administration of ammonium chloride, infants 1 to 16 months of age were similar to older children in their capacity to acidify their urine. The infants had a higher rate of excretion of titratable acid and a lower rate of excretion of ammonium but were similar in their rate of excretion of total hydrogen ion.Bicarbonate titrations performed in infants during the first year of life demonstrated a threshold ranging from 21.5 to 22.5 mmoles per L, maximal rate of reabsorption from 2.6 to 2.9 mmoles per 100 ml glomerular filtrate, and marked titration splay. A nephronic frequency distribution curve of the ratio of glomerular filtration rate to tubular reabsorptive capacity demonstrated both heterogeneity and skewing to the right, suggesting the presence of significant numbers of nephrons with low tubular transport capacity relative to filtration rate.It is suggested that the "physiologic acidosis" of the infant is due neither to a limited renal capacity to excrete hydrogen ion nor to a reduced capacity for reabsorption of bicarbonate, but rather to a low renal plasma bicarbonate threshold. Although the level of the threshold may relate to the kinetics of bicarbonate reabsorption during this period, it appears to be due at least in part to functional and morphologic heterogeneity of nephrons. PMID- 16695920 TI - Studies of the mechanism by which chronic metabolic acidosis augments urinary calcium excretion in man. AB - We carried out clearance studies in nine healthy adults and four patients with hypoparathyroidism before and after inducing stable metabolic acidosis with either NH(4)Cl or acetazolamide. Clearances were repeated in seven normal subjects and three of the patients 3 days after stopping these agents.During acidosis in the normal subjects, serum ultrafilterable calcium concentration rose significantly, but inulin clearance fell to a greater extent, so that the calculated filtered load of calcium fell significantly. Despite this, urinary calcium excretion rose. Urinary calcium excretion remained elevated in the recovery studies when the serum ultrafilterable calcium concentration and filtered load of calcium had returned to control levels. Evidence is presented indicating that the increased calcium excretion which occurred during acidosis and recovery clearances was not due to natriuresis or to increased excretion of complexing anions. The comparable results in the four patients with hypoparathyroidism, two of whom also had hypothyroidism, suggest that the capacity to alter secretion rates of parathyroid hormone, thyrocalcitonin or both is not a critical determinant of the augmented rates of calcium excretion during acidosis.We conclude that metabolic acidosis produces increased urinary calcium excretion by causing decreased renal tubular calcium reabsorption. Evidence is presented which suggests that this is a direct effect of metabolic acidosis on metabolic processes within renal tubular cells. PMID- 16695921 TI - A Low Molecular Weight Immunoglobulin Antigenically Related to 19 S IgM. AB - Sucrose density gradient analysis of the fresh sera of patients with hereditary ataxia telangiectasia, disseminated lupus, and Waldenstrom's macroglobulinemia revealed the presence of an immunoglobulin possessing IgM determinants but having a sedimentation coefficient of approximately 7 S. Bio-Gel chromatography of patients' sera confirmed the presence of two distinct populations of IgM. The low molecular weight IgM possessed incomplete isohemagglutinin activity that was resistant to treatment with reducing agents. Gel diffusion analysis revealed that the 7 S IgM showed immunological identity with both 19 S IgM and the subunits of the 19 S IgM produced by reduction. Approximately 10 to 15% of the patient's total IgM was low molecular weight. Evidence is presented that the 7 S IgM was not produced from the patient's serum 19 S IgM on in vitro incubation. A simple rapid technique is described, using double diffusion in polyacrylamide gels, which permits the determination of low molecular weight IgM in sera and other fluids. Using this technique, the sera of 52 patients with disseminated lupus were surveyed, and 17% of the patients were found to contain low molecular weight IgM. The low molecular weight IgM occurred with particular frequency in male patients with disseminated lupus and in those patients with low or absent serum IgA.Studies of the salivary immunoglobulins of patients with ataxia telangiectasia and disseminated lupus suggest an iverse relationship between the levels of IgA and IgM. In patients lacking salivary IgA, IgM was the major immunoglobulin present. No correlation was observed between salivary immunoglobulin levels and the severity of sinopulmonary infections in these patients. PMID- 16695922 TI - Impaired Lymphocyte Transformation and Delayed Hypersensitivity in Sjogren's Syndrome. AB - In vitro lymphocyte transformation in response to phytohemagglutinin and streptolysin O and in vivo skin sensitization to 2,4-dinitrochloro-benzene has been studied in patients with Sjogren's syndrome and in normal controls of comparable age and sex. Both the in vivo and in vitro responses were significantly impaired in the Sjogren's patients as compared to the controls. This lack of response to mitogenic agents is probably due to an intrinsic defect in the lymphocytes rather than to a serum factor. The abnormalities were less marked in patients whose disease was localized to the parotid and lacrimal glands than in those with generalized disease, i.e., with complicating rheumatoid arthritis or pseudolymphoma. PMID- 16695923 TI - Total and free triiodothyronine in human serum. AB - A reliable method has been developed for the determination of total serum T3, dialyzable fraction (DFT3), and absolute concentration of free T3 (AFT3). Total T3 values (mean +/- SD) were: healthy euthyroid subjects, 0.33 +/- 0.07 mug per 100 ml; hyperthyroid patients, 0.71 +/- 0.1 mug per 100 ml; hypothyroid, 0.10 +/- 0.03 mug per 100 ml. Values (mean +/- SD) for DFT3 in these groups were 0.46 +/- 0.14%, 0.78 +/- 0.17%, and 0.16 +/- 0.08%, respectively. Calculated values for AFT3 were: 1.51 +/- 0.4 mmug per 100 ml, 5.00 +/- 0.6 mmug per 100 ml and 0.24 +/ 0.1 mmug per 100 ml, respectively. Dilution of serum before dialysis lowered estimated DFT3 values. Enrichment of serum with labeled T3 in the range examined did not affect DFT3. However, DFT3 was increased by addition of Merthiolate to serum in concentration 1: 10,000 due to displacement of T3 from thyroxine-binding globulin to albumin. The data suggest that triiodothyronine may play a considerably more important role in normal and pathological physiology, as evidenced by kinetic analysis using these data. A metabolic role for T3 equal to that of T4 is indicated. PMID- 16695924 TI - Variation of proliferative activity in leukemic cell populations of patients with acute leukemia. AB - In 31 children with acute leukemia, the proliferative activity of the leukemic marrow cell population as measured by mitotic and labeling indexes varied widely from patient to patient and from one disease stage to another. Leukemic marrow had a small but statistically significant diurnal variation of proliferative activity. Changes in labeling indexes were directly related to changes in the proportion of large dividing blasts in the marrow. Generation times of dividing leukemic blast cells in 3 patients were similar at diagnosis and in relapse. Changes in proliferative activity of leukemic marrow can be explained by progressive accumulation of nondividing leukemic cells. PMID- 16695925 TI - Multicompartmental analysis of calcium kinetics in normal adult males. AB - This report describes studies of calcium kinetics in ten normal young men. Serum, urinary, and fecal radioactivity was measured from 1 minute to 20 days after intravenous tracer (47)Ca injection, and these results were analyzed jointly with data obtained from a simultaneous metabolic balance study, using digital computer techniques. Surface radioactivity measurements were also obtained to gain further insight into the anatomic correlates of the tracer distribution. The data were satisfied by a model with four exchanging compartments. Series, branching, and mammillary models were analyzed. Several parameters of physiologic interest were independent of the model, but two were dependent on the duration of the study. Individual and mean values for these kinetic analyses are presented with their statistical uncertainties. These studies present detailed analyses in a healthy, normal population and provide a reference for future studies of skeletal metabolism and serum calcium homeostasis. PMID- 16695926 TI - Platelet thrombosthenin: subcellular localization and function. AB - Thrombosthenin, an immunologically distinct contractile protein was isolated in relatively pure form from human platelets. The protein, which was of high molecular weight appeared to be composed of multiple polypeptide subunits, probably polymeric in nature.Thrombosthenin had magnesium-dependent ATPase activisty, releasing an average of 3 mug phosphorus per mg protein in 30 min. After the addition of ATP, there was a reversible alteration in viscosity with calculated ATP sensitivity ranging from 64 to 90%. These biochemical properties of thrombosthenin resemble those of smooth muscle.Specific antisera to thrombosthenin significantly inhibited the ATPase activity of the protein. Clot retraction of recalcified platelet-rich plasma and clot retraction of clotted fibrinogen-platelet mixtures were also inhibited by the antisera. The findings suggest that thrombosthenin is an important component of the clot retraction system.Thrombosthenin was extracted from isolated platelet granule and membrane fractions. The contractile protein derived from the membrane compartment was more active as an ATPase and appeared to be more homogeneous on immunologic analysis. PMID- 16695927 TI - Comparative effects of insulin on adipose tissue segments and isolated fat cells of rat and man. AB - In vitro metabolism of glucose-1-(14)C by adipose tissue into (14)CO(2) and total (14)C lipids in rat and man was compared employing both adipose tissue segments and isolated fat cells prepared from the same donor. In the rat, the basal glucose metabolism and response to insulin decreased with increasing body weight for both adipose tissue segments and isolated cells. Regardless of age, the isolated cells exhibited a persistently higher metabolic activity. Of the parameters selected, conversion to CO(2) was more pronounced than that to lipid.In contrast to the rat, in man adipose tissue segments were more active than isolated cells. In four subjects, the effect of 6, 50, and 400 muU/ml of insulin was analyzed on conversion of glucose-1-carbon to CO(2), long chain fatty acids, and glycerides by adipose tissue segments only. In 17 subjects, glucose oxidation and lipid synthesis by adipose tissue segments and isolated fat cells were measured and showed a definite response to physiological doses of crystalline pork insulin. There was, however, an age dependency, and consistent effects were obtained with 6 muU/ml in children and 50 muU/ml in adults. The responsiveness of human adipose tissue to exogenous insulin in concentrations comparable to those detected in blood reemphasizes the importance of adipose tissue as a major site for fatty acid synthesis. PMID- 16695928 TI - Competitive effects of some cations on active potassium transport in the human red blood cell. AB - The effect of some cations on the active potassium transport system of the human red blood cell has been investigated. At low extracellular potassium concentrations, extracellular sodium competitively inhibits the active potassium influx at all sodium concentrations investigated, and tetraethylammonium behaves in a fashion similar to that of sodium. At low extracellular concentrations of potassium, ammonium at low concentrations at first stimulates the active potassium influx, but at higher concentrations inhibits it. Tetramethylammonium at most slightly stimulates the active potassium influx, and calcium is without effect. The behavior is consistent with a model in which potassium is required at more than one site before transport occurs, and the sites are indistinguishable as far as their behavior toward the ions investigated is concerned. The affinity of the alkali metal cations for the sites appears to be explicable in terms of their physical characteristics. PMID- 16695929 TI - The effects of azathioprine (imuran) on purine synthesis in clinical disorders of purine metabolism. AB - Azathioprine, a purine analogue, significantly suppressed the purine synthesis de novo of two gouty patients manifesting overproduction of uric acid, as well as three of four gouty patients who showed normal uric acid production. This suppression is taken as evidence that phosphoribosyl-pyrophosphate amidotransferase, the rate-controlling step in purine synthesis de novo, has a normal sensitivity to feedback inhibitors in the patients who responded to the drug.Two children afflicted with the familial disorder of hyperuricemia, choreo athetosis, and self-mutilation described by Lesch and Nyhan showed no reduction in the activity of the biosynthetic pathway in response to azathioprine. This inability to respond to azathioprine can be directly related to the absence in these patients of the enzyme hypoxanthine-guanine phosphoribosyltransferase which is required for conversion of the drug or its metabolites to the biochemically active ribonucleotide form. PMID- 16695931 TI - Vascular responses after alpha adrenergic receptor blockade: I. Responses of capacitance and resistance vessels to norepinephrine in man. AB - Experiments were done to test the hypothesis that alpha receptor blockers antagonize more effectively venous than arterial responses to norepinephrine in man.Systemic arterial blood pressure, venous pressure in the forearm, blood flow through the forearm, and the volume of the forearm at a venous pressure of 30 mm Hg were measured using pressure transducers and a mercury strain-gauge plethysmograph. Infusions of norepinephrine into the brachial artery reduced forearm blood flow and venous distensibility without changing arterial pressure. After intraarterial infusion of phentolamine the decrease in venous distensibility during administration of norepinephrine was blocked almost completely whereas the decrease in blood flow through the forearm was not altered.The results indicate that alpha adrenergic receptor blockade can antagonize constriction of capacitance vessels more effectively than constriction of resistance vessels. PMID- 16695930 TI - The Effects of Angiotensin II on Renal Water and Electrolyte Excretion in Normal and Caval Dogs. AB - The effects of intravenous administration of angiotensin II on renal water and electrolyte excretion were examined during hydropenia, water diuresis, and hypotonic saline diuresis in anesthetized normal dogs and dogs with thoracic inferior vena cava constriction and ascites (caval dogs). The effects of unilateral renal artery infusion of a subpressor dose were also examined.During hydropenia angiotensin produced a decrease in tubular sodium reabsorption, with a considerably greater natriuresis in caval dogs, and associated with a decrease in free water reabsorption (T(c) (H(2)O)). Water and hypotonic saline diuresis resulted in an augmented angiotensin natriuresis, with a greater effect still observed in caval dogs. In these experiments free water excretion (C(H(2)O)) was limited to 8-10% of the glomerular filtration rate (GFR), although distal sodium load increased in every instance. In the renal artery infusion experiments a significant ipsilateral decrease in tubular sodium reabsorption was induced, particularly in caval dogs.These findings indicate that angiotensin has a direct effect on renal sodium reabsorption unrelated to a systemic circulatory alteration. The attenuation or prevention of the falls in GFR and effective renal plasma flow (ERPF) usually induced by angiotensin may partially account for the greater natriuretic response in caval dogs and the augmentation during water or hypotonic saline diuresis. However, a correlation between renal hemodynamics and the degree of natriuresis induced was not always present and, furthermore, GFR and ERPF decreased significantly during the intrarenal artery infusion experiments. Therefore, the present experiments indicate that another mechanism is operative in the control of the angiotensin natriuresis and suggest that alterations in intrarenal hemodynamics may play a role.The decrease in T(c) (H(2)O) and the apparent limitation of C(H(2)O) associated with an increase in distal sodium load localize the site of action of angiotensin to the ascending limb of Henle's loop and the proximal tubule. PMID- 16695932 TI - Vascular responses after alpha adrenergic receptor blockade: II. Responses of venous and arterial segments to adrenergic stimulation in the forelimb of dog. AB - The effects of nerve stimulation and of intraarterial injections of norepinephrine on arterial and venous resistances were studied in the perfused forelimb of dog before and after administration of the alpha adrenergic receptor blocker phenoxybenzamine.Pressures were recorded from the perfused brachial artery and a small metacarpal vein in the forepaw. Blood flow to the whole forelimb was maintained constant. Changes in perfusion pressure in the brachial artery reflected primarily changes in arterial resistance and changes in small vein pressure reflected changes in resistance of venous segments downstream from the point of pressure measurement.Alpha receptor blockade reduced vasoconstrictor responses to both nerve stimulation and norepinephrine. Responses to angiotensin, used in these experiments as an internal control, were not blocked consistently in a dose-related manner indicating that the effects of phenoxybenzamine were specific to adrenergic stimuli.Increases in venous pressure in response to norepinephrine and to nerve stimulation were blocked almost completely whereas increases in arterial pressure were reduced only in part by the blocker. The more effective reduction of pressor responses in the small vein was not caused by a passive reduction in blood flow through the paw nor was it caused by a reduction in the concentration of norepinephrine in the venous effluent reaching the venous segments.This differential effect of alpha receptor blockade on increases in venous and arterial resistances may account for the beneficial effect of phenoxybenzamine in shock. PMID- 16695933 TI - Studies on steroid fever: I. Production of leukocyte pyrogen in vitro by etiocholanolone. AB - When a serum-buffer solution of etiocholanolone is incubated with human blood leukocytes in vitro, a pyrogen is released. Like endogenous pyrogen of leukocyte origin, this pyrogen produces prompt monophasic fevers in rabbits, does not induce fever tolerance when given daily, and is inactivated by trypsin. In many respects, the characteristics of the in vitro reaction resemble experimental steroid-induced fever. For example, release of pyrogen varies directly with the concentration of steroid. 4-8 hr of contact between steroid and leukocyte is required for activation of the cell. Rabbit leukocytes are not activated by etiocholanolone. Finally, androsterone, the 5alpha-isomer of etiocholanolone, does not induce pyrogen release in vitro. These studies suggest that experimental steroid fever in man may be mediated by an endogenous pyrogen released from leukocytes. PMID- 16695934 TI - The influence of the parathyroid glands on the hypercalcemia of experimental magnesium depletion in the rat. AB - Magnesium-deficient rats develop significant hypercalcemia, hypophosphatemia, and hyperphosphaturia. These changes suggest a state of hyperparathyroidism. This study examines the regulation of parathyroid gland activity in magnesium deficient rats. Magnesium deficiency was induced in intact and chronically parathyroidectomized animals by feeding them a diet free of this cation. Control animals were pair fed and treated identically except for the inclusion of magnesium in their gavage solution.Magnesium-deficient rats with intact parathyroid glands developed significant hypercalcemia and hypophosphatemia. In addition, the concentration of ionic calcium in plasma was significantly elevated. In contrast, magnesium-deficient parathyroidectomized animals did not have a higher level of calcium in plasma than their nondeficient controls; they developed a decreased concentration of ionic calcium in the absence of a difference in the concentration of phosphate in plasma when compared with appropriate controls. The increased urinary excretion of phosphate was independent of the parathyriod status of the animals.It can be concluded that the hypercalcemia and hypophosphatemia of magnesium deficiency demands parathyroid gland activity and that the regulation of this activity is modified in the magnesium-deficient state to permit the maintenance of an elevated concentration of ionic calcium in plasma. Additional explanations must be found for the hyperphosphaturia. PMID- 16695935 TI - Usefulness of chromic oxide as an internal standard for balance studies in formula-fed patients and for assessment of colonic function. AB - In 35 patients maintained solely on liquid formula diets, chromic oxide has been evaluated as an internal standard for balance studies that require stool collections. In 28 patients the excretion of chromic oxide was ideal: steady states were attained in which mean daily output was 90% (or more) of mean daily intake. In these patients corrections for fecal flow could validly be applied.In patients who excreted the marker ideally, the availability of chromic oxide balance data made possible the calculation of pool sizes and turnover rates of unexcreted intestinal content. These indexes bore little relationship to the usual clinical descriptions of bowel habits. In some patient who had daily bowel movements, pool sizes were very large and daily turnover was small, i.e., a large proportion of the colonic contents was not excreted for surprisingly long periods. It is critically important for investigators to recognize this possibility when carrying out balance studies for fecal constituents that may be altered by bacterial action within the gut lumen: for instance, in 6 patients a significant inverse correlation was found between daily fecal turnover and degradative losses of large amounts of dietary beta-sitosterol.7 of 35 patients failed to attain the ideal steady state of chromic oxide excretion. These patients would not have been singled out if an internal standard had not been used. In such patients balance studies that require analysis of fecal constituents must be interpreted with great caution, since the constituents in question may be handled in the same nonideal fashion as the internal standard. PMID- 16695936 TI - Role of the venous system in baroreceptor-mediated reflexes in man. AB - Although baroreceptor stimulation produced by marked alterations in arterial pressure has been shown to produce reflex changes in venous tone in animals, the effects on venous tone in man of altering arterial pressure within the physiologic range have not been clear. In six subjects, venous tone did not change when mean arterial pressure was raised by 25-40 mm Hg, although heart rate fell reflexly by 40%. Venous tone remained constant in 10 subjects when arterial pressure was lowered. This contrasted to the sustained rise in forearm vascular resistance and the persistent tachycardia that occurred. However, 12 subjects continued to respond to these interventions by transient venoconstriction. To eliminate possible emotional influences on venous tone due to the experimental intervention, venous responses were studied before and during general anesthesia in five of these subjects. In contrast to the response before anesthesia, an equivalent fall in arterial pressure during anesthesia no longer evoked a venoconstrictor response. Venous reactivity and the baroreceptor reflex arc remained intact during anesthesia, since venous tone always rose after a deep inspiration, and tachycardia always accompanied the fall in arterial pressure. It is concluded that changes in arterial pressure in the physiologic range in man do not induce measurable reflex alterations in venous tone, and that the increases sometimes seen with decreases in arterial pressure appear to be due to extraneous psychic factors. PMID- 16695937 TI - The role of adipose cell size and adipose tissue insulin sensitivity in the carbohydrate intolerance of human obesity. AB - Glucose metabolism and insulin sensitivity of isolated human adipose tissue was studied as a function of adipose cell size and number. Glucose metabolism by these tissues was closely related to the number of cells in the fragment, irrespective of cell size. Adipose cells of obese individuals metabolized glucose to carbon dioxide and triglyceride at rates similar to adipose cells of nonobese subjects. In contrast, insulin responsiveness of adipose tissue was dependent upon adipose cell size. The larger its adipose cells the less insulin sensitive was the tissue. Thus, adipose tissue of obese subjects, with enlarged cells, showed a diminished response to insulin. After weight loss and reduction in adipose cell size, insulin sensitivity of the adipose tissue of obese patients was restored to normal. When adipose tissue of obese individuals showed impaired responsiveness to insulin, their plasma insulin levels, after oral glucose, were elevated. Weight loss and reduction in adipose cell size restored plasma insulin concentration to normal, concomitant with the return of normal tissue insulin sensitivity. PMID- 16695938 TI - Cholesterol synthesis in the squirrel monkey: relative rates of synthesis in various tissues and mechanisms of control. AB - Cholesterol synthesis has been extensively investigated in various tissues of lower mammals; however, there is little specific information concerning cholesterologenesis in the primate. Furthermore, experiments in whole animals suggest that important differences may exist in the features of cholesterologenesis in the dog and rat versus the monkey and man. Using the new world squirrel monkey, therefore, we performed the present studies to determine the rates of cholesterologenesis in various tissues per unit weight, to define the relative rates of whole organ synthesis, and to evaluate the operation of control mechanisms in these tissues.In control animals fed a low cholesterol chow diet, the liver and ileum were the two most active sites for cholesterologenesis followed, in order, by the colon, esophagus, and proximal small bowel. Rates of synthesis in 10 other tissues tested were considerably lower than these found in the gastrointestinal tract. When rates of whole organ synthesis were calculated, three tissues, i.e., liver, bowel, and skin, accounted for 92% of the total demonstrable synthetic activity.Following cholesterol feeding utilizing either a solid chow or liquid formula diet, marked suppression of hepatic cholesterologenesis occurred while synthesis in other organs remained essentially unaltered. Similarly, fasting animals for periods up to 96 hr resulted in suppression of synthesis in the liver, but not in various levels of the intestine. Finally, biliary diversion for 48 hr caused a twofold increase in hepatic cholesterologenesis and a six- to eightfold increase in sterol synthesis in the small but not the large intestine. PMID- 16695939 TI - Biosynthetic origin of serum cholesterol in the squirrel monkey: evidence for a contribution by the intestinal wall. AB - The possibility that the intestinal wall serves as a biosynthetic site for serum cholesterol has been examined in two types of studies in the squirrel monkey. First, animals were fed cholesterol in order to inhibit cholesterol synthesis in the liver, and the intestinal lymph ducts were cannulated. After the administration of acetate-2-(14)C it was possible to demonstrate that cholesterol synthesized by the intestinal wall enters intestinal lymph and thereby in the intact animal enters the circulating pool. Second, an attempt to quantitate the significance of this intestinal contribution has been made in animals fed cholesterol-3-(3)H and injected with cholesterol-4-(14)C for long periods. By an application of the technique of analysis utilizing the isotopic steady state we estimated as a minimal value that in the squirrel monkey 1.5-2.0 mg of cholesterol synthesized in the intestinal wall reaches the circulation each day. PMID- 16695940 TI - Respiratory adjustment to chronic metabolic alkalosis in man. AB - This study examined the ventilatory adjustment to chronic metabolic alkalosis induced under controlled conditions in normal human volunteers. Metabolic alkalosis induced by buffers (sodium bicarbonate, trishydroxymethylamine methane) or ethacrynic acid was associated with alveolar hypoventilation, as evidenced by a rise in arterial Pco(2), a fall in arterial Po(2), a reduced resting tidal volume, and a diminished ventilatory response to CO(2) inhalation. Alveolar hypoventilation did not occur when metabolic alkalosis was induced in the same subjects by thiazide diuretics or aldosterone despite comparable elevations of the arterial blood pH and bicarbonate concentration.The different ventilatory responses of the two groups could not be ascribed to differences among individuals comprising each group, pharmacological effects of the alkalinizing agents, differences in the composition of the lumber spinal fluid, changes in extracellular fluid volume, or sodium and chloride balance.The differences in ventilatory adjustments were associated with differences in the patterns of hydrogen and potassium ion balance during the induction of alkalosis. Alveolar hypoventilation occurred when hydrogen ions were buffered (sodium bicarbonate, trishydroxymethylamine methane) or when renal hydrogen ion excretion was increased (ethacrynic acid). Alveolar hypoventilation did not occur when induction of similar degrees of extracellular alkalosis was accompanied by marked potassium loss and no demonstrable increase in external hydrogen loss (thiazides and aldosterone).These observations suggest that respiratory depression does not necessarily accompany extracellular alkalosis but depends on the effect of the mode of induction of the alkalosis on the tissues involved in the control of ventilation. PMID- 16695941 TI - Estradiol metabolism in cirrhosis. AB - Abnormal estrogen metabolism has been found in cirrhosis after administration of intravenous tracers of estradiol-(3)H to 6 patients and 23 healthy controls. The major abnormalities observed involved estrogen metabolites other than the 3 "classic" ones, i.e., estrone (E1), estradiol (E2), and estriol (E3). Urinary recovery of radioactivity was regularly elevated in the patients, to an average of 71% of the dose compared to 51% in normals. This is considered to reflect the component of intrahepatic cholestasis in cirrhosis. The per cent dose recovered as urinary glucosiduronates (42%) was normal in cirrhotics in contrast to impaired glucuronidation of cortisol metabolites in this disease. E1 and E2 were present in normal amounts, and E3 was slightly elevated to 21% of the extract compared to 14% in controls. There were strikingly decreased excretion of 2 hydroxyestrone (3% compared with normal 20%) and 2-methoxyestrone (2% compared with 5%) and increased excretion of 16alpha-hydroxyestrone (12% compared with normal 6%). Thus cirrhosis, too, is characterized by the reciprocal relationship between decreased 2-hydroxylation and increased 16alpha-hydroxylation previously described in hypothyroidism and male breast cancer. However, unlike these latter, the increase of 16alpha-hydroxy metabolites was less than the decrease of 2 hydroxy metabolites. The data indicate clearcut impairment of 2-hydroxylation, suggestive impairment of 16alpha-hydroxylation, and a definite depression of the reaction 16alpha-hydroxyestrone-->estriol, the latter finding so far unique to cirrhosis. Demonstration of abnormal peripheral metabolism of estrogen in cirrhosis provides a new approach to the origin of the hyperestrogenic syndrome in this disease. PMID- 16695942 TI - Micropuncture study of nephron function in the rhesus monkey. AB - The function of the proximal and distal tubule was studied in the rhesus monkey during antidiuresis and during the diuresis after furosemide administration (during which extracellular fluid volume was maintained).In the proximal tubule, fluid to plasma ratios for sodium, potassium, and osmolality approximated unity. During antidiuresis, about 30% of the filtered water remained at the end of the accessible portion of this segment (92% of length). Fluid was hypotonic to plasma throughout the distal tubule. 25% of the filtered water was present in the early distal tubule. Small but significant water reabsorption (about 8% of filtered) occurred in remainder of this segment. Tubule fluid to plasma potassium concentration ratios tended to increase along the distal tubule, and the amount of potassium, relative to the amount filtered, increased from 13% in the early portion of this segment to 26% in the late portion.After furosemide was administered animals excreted about one-third of the filtered sodium and water. Despite this diuresis, electrolyte and water reabsorption along the proximal tubule did not differ from values obtained in control animals. Osmolality and sodium concentration of fluid from the distal tubule approached those of plasma. 22% of the filtered sodium (twice the control values) reached the distal tubule, whereas the fraction of filtered water remaining was only slightly increased. These findings indicate that, after the administration of this drug, inhibition of sodium reabsorption occurred in the water-impermeable segment of the nephron, rather than in the proximal tubule. After furosemide administration, all tubule fluid to plasma potassium concentration ratios in the distal tubule were equal to or greater than one, suggesting inhibition of active potassium reabsorption at or prior to this site.Fluid to plasma bicarbonate concentration ratios from the midportion of the proximal tubule were consistently less than one in normal monkeys. After acetazolamide was administered, the bicarbonate concentration of samples of tubule fluid recollected from these same sites was the same as, or higher than in plasma. This fact demonstrates the inhibition of bicarbonate reabsorption in this portion of the tubule. PMID- 16695943 TI - The effect of steady-state increases in systemic arterial pressure on the duration of left ventricular ejection time. AB - The effect of steady-state increases in systemic arterial pressure on the duration of left ventricular ejection time was studied in 11 normal male subjects. Methoxamine, a pressor amine of predominantly vasoconstrictor activity but lacking significant inotropic effect, was administered intravenously resulting in an average increase in mean arterial pressure of 27 mm Hg. Heart rate was held constant by high right atrial pacing, and there was no significant change in cardiac output. During methoxamine infusion, when stroke volume, heart rate, and inotropic state were held constant, left ventricular ejection time increased as mean arterial pressure increased. There was a highly significant correlation between the increase in mean systolic blood pressure and the prolongation of left ventricular ejection time (r = 0.870). In one subject, an increase in mean systolic pressure of 75 mm Hg prolonged left ventricular ejection time 55 msec, producing paradoxical splitting of the second heart sound. The prolongation of left ventricular ejection time during infusion was not blocked by the prior intravenous administration of atropine sulfate or propranolol hydrochloride, thus ruling out both vagal inhibition of the left ventricle and reflex withdrawal of sympathetic tone as its cause. In three subjects, left ventricular end diastolic pressure was measured and found to be significantly increased. This finding suggests that the normal left ventricle maintains a constant stroke volume in the presence of an increased pressure load by the Frank Starling mechanism. This study concludes that arterial pressure must be included as a prime determinant of left ventricular ejection time along with stroke volume, heart rate, and inotropic state in intact man. PMID- 16695944 TI - Estimation of thyroxine distribution in man. AB - A group of 13 normal subjects were evaluated for their extrathyroidal thyroxine distribution. The method employed the measurement of the acute plasma disappearance of a thyroxine-(131)I tracer and its concomitant uptake into the liver and forearm. The analysis of these parameters allowed the theoretical construction of a four compartmental mathematical model system comprised of the plasma, extracellular fluid, hepatic, and extrahepatic thyroxine pools. The results of this analysis revealed that the exchange of thyroxine from the plasma into the hepatic and extrahepatic cellular fluid spaces appeared, in general, to be rapid, while the uptake into the extrahepatic tissues was relatively slow. The calculated distribution of thyroxine at equilibrium was estimated to be 14% in liver, 34% in extrahepatic tissues, and 26% each in the plasma and extracellular fluid pools in this group of normal subjects. PMID- 16695945 TI - Metabolic clearance and production rates of human luteinizing hormone in pre- and postmenopausal women. AB - Metabolic clearance rates and production rates of human luteinizing hormone (HLH) were determined in pre- and postmenopausal women by the constant infusion technique. Highly purified HLH-(131)I was infused into the fasting subjects at a constant rate. Serial plasma samples were obtained and the radioactive hormone was precipitated by a double antibody technique. Plasma HLH-(131)I levels reached equilibrium by 4 hr after the infusion started. Metabolic clearance rates were: 24.4 +/- 1.8 (mean +/- SE) ml/min in five normal premenopausal women; 23.3 +/- 1.1 ml/min in five normal women taking norethinodrel and mestranol; and 25.6 +/- 4.1 ml/min in four postmenopausal women. Endogenous plasma HLH levels measured in the same subjects by radioimmunoassay immediately before infusion were 32.0 +/- 9.6 mU/ml in the normal women, 16.8 +/- 3.2 mU/ml in the women on oral contraceptives, and 99.2 +/- 23.2 mU/ml in the postmenopausal women. The corresponding HLH production rates were: 734 +/- 170 mU/min in the normal women: 387 +/- 86 mU/min in the women on norethinodrel and mestranol; and 2400 +/- 410 mU/min in the postmenopausal women. The metabolic clearance rate did not change after ovariectomy in one women, but the production rate rose from 583 to 1420 mU/min. Based on previously reported bioassay values for pituitary content and urinary excretion of HLH, the estimated turnover of HLH in the pituitary is about once per day and less than 5% of the total HLH produced appears in the urine in a biologically active form. PMID- 16695946 TI - Reduced nicotinamide adenine dinucleotide and the reduction of oxidized glutathione in human erythrocytes. AB - The ability of reduced nicotinamide adenine dinucleotide (NADH), generated through the activity of lactic acid dehydrogenase, to support the reduction of endogenous oxidized glutathione in intact human erythrocytes and in hemolysates was investigated. Rapid initial oxidation of endogenous reduced glutathione was effected with methyl phenylazoformate. Freshly obtained normal erythrocytes and erythrocytes deficient in glucose-6-phosphate dehydrogenase activity were unable to regenerate reduced glutathione upon incubation with lactate. Only normal erythrocytes were capable of reducing oxidized glutathione after preincubation with glucose, inosine, or a medium which promoted the synthesis of increased amounts of intracellular NAD. This regeneration of reduced glutathione could be explained by the generation of reduced nicotinamide adenine dinucleotide phosphate through the metabolism of accumulated phosphorylated intermediates of glycolysis. Hemolysates prepared from both normal erythrocytes and from erythrocytes deficient in glucose-6-phosphate dehydrogenase activity were able to reduce oxidized glutathione in the presence of added lactate and NAD. The results obtained indicated either an inability of the intact erythrocyte to utilize the NAD at the concentrations attained or an altered behavior of the system for the regeneration of reduced glutathione after lysis of the cell. PMID- 16695947 TI - The phospholipid requirement of tissue factor in blood coagulation. AB - Using a coagulation assay specific for tissue factor, we found that removal of 95% of the tissue factor-phospholipid resulted in a loss of 98% of its biological activity. The activity could be restored, with yields in excess of 100% by combining the extracted tissue factor with either mixed brain phospholipids or highly purified phospholipids. Phosphatidylethanolamine was the most active, followed by phosphatidylcholine. Phosphatidylserine, phosphatidylinositol, and sphingomyelin had little or no activity. In addition, a requirement for unsaturation and the presence of two fatty acids was demonstrated. The activity of phosphatidylcholine was also dependent on the presence of the base. Furthermore, it was shown that activity was not a function of binding of phospholipids to tissue factor, as both active and inactive lipids were equally bound. PMID- 16695948 TI - Regional distribution of pulmonary ventilation and perfusion in elderly subjects. AB - Using radioactive xenon, we measured the regional distribution of pulmonary ventilation and blood flow in six normal men, whose ages ranged between 65 and 75 yr. The measurements were made in the standing position. The static volume pressure relation of the lungs was also measured in five of the subjects. The results indicate that by comparison with normal young men: (a) Blood flow to the upper lung zones was increased, although it still remained predominant in the lower zones. (b) Ventilation distribution during a vital capacity inspiration was similar to that seen in young subjects. (c) In five of the six elderly subjects, however, the distribution of ventilation in the resting tidal volume range was not preferential to the lower zones as it was in young men. This was probably caused by airway closure in the lower lung zones. The elderly subjects thus exhibit during normal tidal volume breathing a ventilation distribution pattern similar to that observed in young subjects when breathing at low lung volumes, i.e., near residual volume. This difference is probably due to the combined effect of the loss in elastic recoil of the lungs observed in the elderly subjects and of a decreased resistance to collapse of the aged airways. These findings suggest that in the elderly subjects there is a significant regional ventilation-perfusion impairment during quiet breathing, which may explain in part the reported increase in alveolar-arterial oxygen difference with advancing age. PMID- 16695949 TI - Metabolic clearance rates and interconversions of estrone and 17beta-estradiol in normal males and females. AB - The continuous infusion of (3)H-6,7-estrone and (3)H-6,7-estradiol has been used to study the metabolic clearance rate (MCR), the interconversions, and the red cell uptake of these steroids in normal males and females. The whole blood MCR of estrone is 1,990 +/- 120 liters per day/m(2) (SE) in males and 1,910 +/- 100 liters per day/m(2) in females. The whole blood MCR of estradiol is 1,600 +/- 80 liters per day/m(2) in males and 1,360 +/- 40 liters per day/m(2) in females. The values in females do not vary significantly when studied in the follicular or luteal phase of the cycle. At least 35% of the total estrone metabolism in both sexes is extrasplanchnic and at least 25% of the total estradiol metabolism in males, and 15% in females is extrasplanchnic. The [rho](BB) (2,1) [transfer constant of estradiol to estrone, which is equivalent to the fraction of the precursor (estradiol) converted to the product (estrone) when both the infusion of the precursor and the measurement of the product are in peripheral blood] is 15%; and the [rho](BB) (1,2) [transfer constant of estrone to estradiol, which is equivalent to the fraction of the precursor (estrone) converted to product (estradiol) when both the infusion of the precusor and the measurement of the product are in peripheral blood] is 5% in both males and females. Our findings concerning the radioactivity in whole blood, as measured by our procedure, were the following: 15-20% of estrone in both sexes and 15% of estradiol in males is associated with red cells. Only 2% of the whole blood radioactivity of estradiol in females is associated with red cells. Changes in the distribution of radioactivity between plasma and red cells will influence the MCR as calculated from plasma, but not as calculated from whole blood. PMID- 16695950 TI - On the shear rate dependence of red cell aggregation in vitro. AB - Non-Newtonian viscosity of blood, i.e., the rise in apparent viscosity at low flow, was believed to be a result of reversible aggregation of red cells at low velocity gradients (shear rate). By making a cone-plate viscometer transparent, direct observation was made possible of the blood flowing under defined shear rates. Red cell aggregates, occurring in all cases at low flow, were reversibly dispersed by increasing the shear rate. This behavior was independent of the addition of anticoagulants, but it could be altered by changing the plasma protein composition. Red cells in serum did not form aggregates; such nonaggregating samples did show an increase in viscosity at low shear rates. Since the sedimentation rate can be influenced by many parameters, it is not reliable in describing red cell aggregation. Aggregation of red cells is linked with a marked separation of plasma and cells. Such a separation is of considerable influence on cone-plate viscometry. PMID- 16695951 TI - Presidential address. PMID- 16695952 TI - Brain metabolism in uremic and adenosine-infused rats. AB - Analyses of nucleotides and glycolytic intermediates were performed on perchlorate extracts of blood and quick-frozen brain from rats nephrectomized 48 hr previously, and from rats infused for 6 hr with adenosine or AMP. Blood nucleotides of acutely uremic rats were normal. Uremic brain showed an increase of creatine phosphate (CP), ATP, and glucose with a corresponding decrease in creatine, ADP, AMP, and lactate. Other nucleotide triphosphates were increased, but total adenine nucleotide in brain was unchanged. Uremic brain failed to use ATP or produce ADP, AMP, and lactate at normal rates when subjected to the stress of ischemic anoxia. Although levels of cation responsive ATPase in extracts of uremic brain were normal, the inhibition of glycolysis in the intact brain appeared to be due to a failure of ATP hydrolysis (a diminished ATPase activity). Adenosine infusion produced mild azotemia, marked hyperglycemia, an increase in blood ATP, and an increase in total blood adenine nucleotide. Brain from rats infused with adenosine or AMP also had high levels of ATP, creatine phosphate, and glucose, whereas levels of ADP, AMP, and lactate were low. However these brains responded with normal use of ATP and normal production of lactate when stimulated by ischemic anoxia. PMID- 16695953 TI - The regulation of antidiuretic hormone release in man: I. Effects of change in position and ambient temperature on blood ADH levels. AB - It has been postulated that alterations in the intravascular distribution of blood affect antidiuretic hormone (ADH) secretion in man. The studies reported here were designed to alter blood distribution by thermal and by positional change to test this thesis.HUMAN BLOOD ADH LEVELS HAVE BEEN SHOWN TO VARY WITH POSITION: a mean value of 0.4 +/- 0.6 (SD) muU/ml was obtained while the subject was supine, a value of 1.4 +/- 0.7 muU/ml while sitting, and 3.1 +/- 1.5 muU/ml while standing. In 79 control subjects, sitting comfortably for 30 min in a normal environment, a blood ADH level of 1.65 +/- 0.63 muU/ml was found. It is suggested that subjects assume this position during experiments in which blood is drawn for measurement of ADH levels.In eight seated subjects the ADH level rose from 1.6 +/- 0.4 to 5.2 +/- 0.8 muU/ml after a 2 hr exposure at 50 degrees C and fell to 1.0 +/- 0.26 muU/ml within 15 min at 26 degrees C.Six subjects with a mean ADH level of 2.2 +/- 0.58 muU/ml sat quietly in the cold (13 degrees C) for 1 hr, and the ADH level fell to 1.2 +/- 0.36 muU/ml. After 15 min at 26 degrees C, the level rose to 3.1 +/- 0.78 muU/ml. The serum sodium and osmolal concentrations remained constant during all studies.Water, sodium, and total solute excretion decreased during exposure to the heat, whereas the urine to plasma (U/P) osmolal ratio increased. During cold exposure, water, sodium, and total solute excretion increased, and there was a decrease in the U/P osmolal ratio.These data are interpreted as indicating that changes in activity of intrathoracic stretch receptors, in response to redistribution of blood, alter ADH secretion independently of changes in serum osmolality. The rapidity of change of blood ADH concentration indicates a great sensitivity and a prime functional role for the "volume receptors" in the regulation of ADH secretion. PMID- 16695954 TI - Attempt to transfer contact hypersensitivity in man with dialysate of peripheral leukocytes. AB - In humans delayed hypersensitivity to tuberculin and coccidiodin can be transferred from a skin-test positive donor to a previously nonreactive recipient with a dialyzable fraction of donor peripheral leukocytes. The mechanism by which transfer is effected by this relatively low molecular weight material is unknown. Should transfer with dialysate be accomplished with hypersensitivity to low molecular weight artificial antigens, further characterization of "transfer factor" might be facilitated.A maximal degree of contact sensitivity to a simple chemical, dinitrochlorobenzene, was produced in tuberculin-sensitive human volunteers, who were then used as donors. Seven of the eight donors reacted with 15 mm or more of induration to 0.1 mug of tuberculin, purified protein derivative, while recipients did not react to 5 mug. The dialyzable fraction of peripheral leukocytes from 500 ml of donor blood was injected into each of eight recipients in an attempt at simultaneous transfer of systemic sensitivity to both tuberculin and dinitrochlorobenzene. Transfer of tuberculin sensitivity was accomplished with four of seven dialysates from tuberculin-positive donors but in none of the eight attempts was contact sensitivity transferred with the same dialysates.The results indicate that tuberculin sensitivity and contact type sensitivity to a simple chemical differ in respect to the behavior of transfer factor and that other models are necessary if dialyzable transfer factor is to be studied with nonbacterial antigens. PMID- 16695955 TI - Metabolic and calcium kinetic studies in idiopathic hypercalciuria. AB - Calcium balances and calcium kinetic studies using (47)Ca were performed in nine male patients with idiopathic hypercalciuria and in three normal male subjects. A sharp reduction in calcium intake in eight patients with idiopathic hypercalciuria caused a decrease in urinary calcium excretion, the latter remaining elevated above that reported for normal subjects on a low calcium diet. The hypercalciuric patients had an enlarged miscible calcium pool size, an increased calcium turnover rate, increased bone formation and bone resorption rates, and an elevated true intestinal calcium absorption rate, the increase of the latter three parameters being proportional to the increase of the turnover rate. The fraction of the calcium turnover rate excreted in the urine was elevated whereas that constituted by the endogenous fecal calcium excretion was decreased. Arguments are presented for the concept that the primary abnormality in idiopathic hypercalciuria is neither renal calcium hyperexcretion nor intestinal calcium hyperreabsorption, but a more fundamental disturbance in calcium metabolism of as yet unknown cause, leading to a high calcium turnover. PMID- 16695956 TI - Studies in clinical shock and hypotension: VI. Relationship between left and right ventricular function. AB - Left ventricular end diastolic (LVEDP) and mean right atrial (RAP) pressures were recorded simultaneously in 30 patients with shock (14 acute myocardial infarction, 10 acute pulmonary embolism or severe bronchopulmonary disease, and 6 sepsis). Myocardial infarction was characterized by a predominant increase in LVEDP, pulmonary disease by a predominant increase in RAP, and sepsis by a normal relationship between LVEDP and RAP. In all three groups a significant positive correlation was noted between RAP and LVEDP, with the regression line in cor pulmonale deviated significantly toward the RAP axis and the regression line in myocardial infarction exhibiting a zero RAP intercept at an elevated LVEDP.Low cardiac outputs with elevated LVEDP in myocardial infarction indicated severe left ventricular failure. Low outputs with elevated RAP in cor pulmonale were consistent with right ventricular overload. Although cardiac outputs often were normal in sepsis, low outputs with elevated cardiac filling pressures in some patients were consistent with a hemodynamic or humoral-induced generalized depression of cardiac performance.Vasoconstrictor and inotropic drugs often produced a functional disparity between the two ventricles, with the gradient between LVEDP and RAP increasing, apparently because of an increase in left ventricular work or an inadequacy of left ventricular oxygen delivery. Acute plasma volume expansion with dextran in patients with pulmonary vascular disease resulted in a somewhat more rapid rise in RAP than in LVEDP. In septic and myocardial infarction shock, however, LVEDP and RAP usually rose proportionally, with the absolute rise of LVEDP surpassing that of RAP. Although the absolute level of the central venous pressure thus may not be a reliable indicator of left ventricular function in shock, changes in venous pressure during acute plasma volume expansion should serve as a fairly safe guide to changes in LVEDP. PMID- 16695957 TI - Quantitative immunology of immune hemolytic anemia: II. The relationship of cell bound antibody to hemolysis and the effect of treatment. AB - The concentration of cell-bound and serum antibody was determined in a series of patients with warm antibody immune hemolytic anemia by determining the amount of C[unk]1 fixed to the cells by anti-IgG. This was compared to the rate of hemolysis as determined by hemoglobin concentration and reticulocyte count, or the endogenous production of carbon monoxide. The rate of hemolysis was, in general, proportional to the concentration of cell-bound antibody. In splenectomized patients, the rate of hemolysis was very much less than in unsplenectomized patients for a given concentration of cell-bound antibody. When prednisone was given, three effects were noted: (a) at high doses of drug, the concentration of cell-bound antibody decreased rapidly and the concentration of serum antibody increased, suggesting that the affinity of antibody for antigen had been altered; (b) in patients achieving remission, the concentration of serum antibody fell to low levels but rose again if the dose of prednisone was insufficient; (c) in one patient, prednisone appeared to inhibit sequestration of highly sensitized cells. PMID- 16695958 TI - The interaction of human macrophages and lymphocytes in the phytohemagglutinin stimulated production of interferon. AB - In studies of 13 normal adults to determine the blood cell types responsible for interferon production induced by phytohemagglutinin, the following observations were made. (a) In cultures containing 96-100% pure macrophages derived from blood monocytes, no interferon was detected in either the presence or the absence of phytohemagglutinin for up to 92 hr. (b) In cultures of 99.5-100% pure lymphocytes, low levels of interferon were detected in the presence, but not in the absence, of phytohemagglutinin. (c) An average fivefold increase in interferon titers occurred when pure lymphocytes were combined with the macrophages in culture with phytohemagglutinin. The peak response of interferon occurred at 68 hr after the initiation of the combined cultures. For maximum response, phytohemagglutinin was required for the duration of the culture, and both cell types in association were necessary. Medium from phytohemagglutinin stimulated macrophages or lymphocytes could not substitute for the corresponding intact cell. However, frozen-thawed macrophages in combination with lymphocytes and phytohemagglutinin produced an intermediate interferon response. An increase in either cell type produced an increased response in the range studied: lymphocytes, 0.45-1.8 x 10(6) per ml; and macrophages, 0.5-2.1 x 10(5) per ml. Syngeneic fibroblasts, HeLa cells, or mouse macrophages could not substitute for the human macrophages in the combined cultures with phytohemagglutinin. (d) Although all cultures producing interferon showed some degree of transformation (thymidine-(3)H incorporation into deoxyribonucleic acid), no direct correlation between the degree of phytohemagglutinin-induced lymphocyte transformation and the interferon titers was observed.The demonstration of macrophage-lymphocyte interaction in the production of interferon is of interest in view of the known interrelationship of these same cell types in antibody synthesis and cellular immunity. PMID- 16695960 TI - 1972 program american society for clinical investigation, inc. 64th annual meeting. PMID- 16695959 TI - Atlantic city meetings 1971. PMID- 16695961 TI - Parathyroid Hormone in Human Plasma: IMMUNOCHEMICAL CHARACTERIZATION AND BIOLOGICAL IMPLICATIONS. AB - Antigenic recognition of four anti-bovine parathyroid hormone antisera was characterized by their reactivity with bovine hormonal fragments (1-34, 1-13, 14 34, 19-34, 53-84) and human hormone extracted from parathyroid adenomas. All antisera were found to have antibody populations which recognized more than one antigenic determinant and all antisera differed in their specificity and reactivity for the fragments of bovine hormone. By modification of two antisera, GP-1 and GP-133, by preincubation with excess concentrations of 1-34 or 53-84 fragments, antigenic recognition was restricted to defined regions of the hormonal sequence.When assays using these modified antisera were applied to the study of hormones extracted from glands, greater immunochemical similarities were seen between bovine and human parathyroid hormone using assays that were specific for the measurement of amino-terminal portions of the hormones than of the carboxy-terminal portions.When assays using these antisera were applied to the study of endogenous parathyroid hormone in human plasma, the immunoreactive hormone in the general circulation was shown to substantially lack an amino terminal portion of the sequence of the intact hormone, including an antigenic determinant requiring all or some of the 14-19 region. This deletion accounts, at least in part, for the immunochemical heterogeneity of plasma parathyroid hormone in man. Radioimmunoassay of fractions of peripheral plasma subjected to gel filtration confirms that the dominant form of the immunoreactive hormone in the general circulation of man is a hormonal fragment that is totally devoid of amino terminal reactivity. Because of this deletion, it can be concluded that most of the immunoreactive parathyroid hormone in the general circulation of man must be biologically inactive. PMID- 16695962 TI - Differences in Cyclic AMP Changes after Stimulation by Prostaglandins and Isoproterenol in Lymphocyte Subpopulations. AB - Various lymphocyte populations have been studied for their content in cyclic adenosine 3',5'-monophosphate (cAMP) before and after stimulation by isoproterenol and prostaglandin E(1) (PGE(1)). Basal cAMP levels vary among lymphocytes according to their origin: peripheral blood lymphocytes show high cAMP level while spleen and lymph node cells and thymocytes show lower levels. Thymocytes are extremely sensitive to the stimulating effects of isoproterenol and PGE(1), much more than spleen and lymph node or peripheral blood cells. Corticoresistant thymocytes are less sensitive to isoproterenol stimulation than normal thymocytes, but are significantly more sensitive than peripheral thymus derived (T)-cells. Studies using bone-marrow-derived (B) or T cell depletion with anti-immunoglobulin-coated columns and antitheta serum (AthetaS) indicate that lymph node B cells synthesize more cAMP in the presence of isoproterenol than T cells. However, this difference between T and B cells has not been found in spleen cells. PMID- 16695963 TI - Fitzgerald Trait: Deficiency of a Hitherto Unrecognized Agent, Fitzgerald Factor, Participating in Surface-Mediated Reactions of Clotting, Fibrinolysis, Generation of Kinins, and the Property of Diluted Plasma Enhancing Vascular Permeability (PF/Dil). AB - The prolonged partial thromboplastin time observed in the plasma of a 71-yr-old asymptomatic man was related to the deficiency of a hitherto unrecognized agent. The patient's plasma also exhibited impaired surface-mediated fibrinolysis and esterolytic activity and impaired generation of kinins and of the property enhancing vascular permeability designated PF/Dil. The patient's plasma contained normal amounts of all known clotting factors except Fletcher factor (a plasma prekallikrein) which was present at a concentration of 10-15% of pooled normal plasma. Fletcher trait plasma, however, contained normal amounts of the agent missing from the patient's plasma and corrected the defects in clotting, fibrinolysis, and vascular permeability. Fletcher trait plasma was less effective in correcting generation of kinins and esterolytic activity, presumably because of the patient's partial deficiency of prekallikrein. The site of action of the factor deficient in the patient's plasma appeared to be subsequent to the activation of Hageman factor and plasma prekallikrein. A fraction of normal plasma, devoid of other clotting factors, corrected the defect in clotting in the patient's plasma; a similar fraction of the patient's plasma did not correct this abnormality. No evidence yet exists pointing to the familial nature of the patient's defect. Tentatively, the patient's disorder may be referred to by his surname as Fitzgerald trait, and the agent apparently deficient in his plasma as Fitzgerald factor. PMID- 16695964 TI - The Use of a Helium-Oxygen Mixture during Maximum Expiratory Flow to Demonstrate Obstruction in Small Airways in Smokers. AB - We measured the response to breathing a mixture of 80% helium and 20% oxygen (He) during a maximum expiratory flow-volume (MEFV) maneuver in 66 nonsmokers and 48 smokers, aged 17-67. All of the subjects studied had (forced expiratory volume in 1 s/forced vital capacity [FEV(1.0)/FVC]) x 100 of greater than 70%. While the flow rates of the smokers were within +/-2 SD of those of the nonsmokers at 50% VC (Vmax(50)), both groups showed a reduction in flow with age (nonsmokers: r= 0.34, P<0.01; smokers r=-0.52, P<0.001). Nonsmokers showed no significant reduction with age in response to breathing He, while smokers showed a marked reduction with age (r=-0.63, P<0.001 at Vmax(50)). We also measured the lung volume at which maximum expiratory flow (Vmax) while the subject was breathing He became equal to Vmax while he was breathing air, and expressed it as a percent of the VC. This was the most sensitive method of separating smokers from nonsmokers. These results indicate that the use of He during an MEFV maneuver affords sufficient sensitivity to enable detection of functional abnormalities in smokers at a stage when Vmax while they are breathing air is normal. PMID- 16695966 TI - Retraction. On the binding of tumor necrosis factor (TNF) to heparin and the release in vivo of the TNF-binding protein I by heparin. PMID- 16695965 TI - Jejunal perfusion of simple and conjugated folates in tropical sprue. AB - Absorption of labeled simple 3',5',9'-(3)H pteroylmonoglutamate, ([(3)H]PG-1) and conjugated pteroyl-mu[(14)C]glutamyl-gamma-hexaglutamate, ([(14)C]PG-7) folates was assessed in six patients with tropical sprue, before and after 6 mo of treatment, utilizing jejunal perfusion and urinary recovery techniques. Degradation products of [(14)C]PG-7 which were produced during perfusion were identified by DEAE-cellulose column chromatography. Jejunal mucosal activities of folate conjugase, lactase, sucrase, and maltase were measured in every patient. Malabsorption of both [(3)H]PG-1 and [(14)C]PG-7 was found in every untreated patient, with significant improvement after therapy. The urinary excretion of (3)H and (14)C paralleled the luminal disappearance of both isotopes. The chromatographic patterns of intraluminal degradation products of [(14)C]PG-7 obtained during perfusion did not differ from those previously found in normal subjects and were similar in studies performed before and after treatment. The activity of folate conjugase was increased in the mucosa of the untreated patients when compared to the post-treatment levels while the activities of mucosal lactase, sucrase, and maltase were originally low and increased significantly after therapy. These observations suggest that folate conjugase originates at a different mucosal locus than the brush border disaccharidases, and are consistent with previous evidence that folate conjugase is an intracellular enzyme. The present studies have demonstrated unequivocal malabsorption of both simple and conjugated folates in tropical sprue. In tropical sprue, folate malabsorption is the reflection of impaired folate transport and not of impaired hydrolysis. PMID- 16695967 TI - A welcome to the CMP edition. PMID- 16695968 TI - Immunohistochemically detectable p53 and mdm-2 oncoprotein expression in colorectal carcinoma: prognostic significance. AB - Aims-To investigate the correlation between the expression of the p53 and mdm-2 oncoproteins and to assess their prognostic value in colorectal cancer.Methods Using a polyclonal (CM1) and a monoclonal antibody directed against p53 and mdm 2, respectively, these oncoproteins were stained immunohistochemically in 109 colorectal adenocarcinomas.Results-p53 was detected in less than 10% of tumour cells in 11 of 109 adenocarcinomas, in 10-50% of tumour cells, in 17 of 109 adenocarcinomas, and in more than 50% of tumour cells in 32 of 109 adenocarcinomas. Expression of mdm-2 was detected in 22 of 109 (20%) cases investigated, of which 19 showed concomitant p53 expression. In most cases mdm-2 immunoreactivity was strongly associated with a small proportion of p53 positive tumour cells. Both p53 and mdm-2 expression lacked statistical significance when correlated with common staging and grading parameters.Conclusions-Detection of p53 and mdm-2 oncoprotein expression, detected using immunohistochemistry, is of no prognostic value in colorectal cancer. However, the close correlation between mdm-2 immunoreactivity and the proportion of p53 positive cells provides further evidence that the mdm-2 gene product interacts with p53 protein. PMID- 16695969 TI - Expression of bcl-2 and p53 proteins in nasopharyngeal carcinoma. Absence of correlation with the presence of EBV encoded EBER1-2 transcripts and latent membrane protein-1. AB - Aims-To investigate the immunohistochemical expression of bcl-2 and p53 proteins in nasopharyngeal carcinomas in relation to the expression of the Epstein-Barr virus (EBV) encoded EBER messenger RNAs (mRNAs) and latent membrane protein-1 (LMP-1).Methods-Formalin fixed, paraffin wax embedded tissue from 44 nasopharyngeal carcinomas (NPCs) was stained by immunohistochemistry for p53, bcl 2 and LMP-1 proteins and by RNA in situ hybridisation for EBER mRNAs.Results-The tumours were divided histologically into 13 cases of keratinising squamous cell NPC (KNPC), 15 cases of non-keratinising squamous cell NPC (NKNPC) and 16 cases of undifferentiated NPC (UNPC). Bcl-2 expression was observed in five of 15 NKNPC cases and in six of 16 UNPC cases; p53 expression was observed in one of 13 KNPC, two of 15 NKNPC and four of 16 UNPC cases. EBER 1-2 transcripts were detected in five of 15 NKNPC and nine of 16 UNPC cases, while LMP-1 expression was observed in one of 16 UNPC cases. All 13 KNPCs were EBV and bcl-2 negative. No correlation was found between the presence of EBER 1-2 transcripts and the detection of bcl-2 or p53 proteins, or both, in NPC cells.Conclusions-The expression of bcl-2 and p53 proteins may be associated with the level of the tumour cell differentiation in NPC. In addition, in view of the important role of the bcl-2 protein in the inhibition of apoptosis, the expression of bcl-2 protein may contribute to tumour cell survival in a proportion of NPCs. Furthermore, in the light of previous findings that the p53 gene in most UNPCs is in the wild-type configuration, mechanisms other than mutation may be responsible for stabilisation of the p53 protein in UNPCs. PMID- 16695970 TI - Determination of cell proliferation. PMID- 16695971 TI - Effects of hepatocyte growth factor on differentiation and cMET receptor expression in the promyelocytic HL60 cell line. AB - Aim-To determine the effects of hepatocyte growth factor (HGF) on myeloid cell differentiation and cMET expression using the promyelocytic HL60 cell line.Methods-HL60 cells cultured with purified recombinant HGF, dimethyl sulphoxide (DMSO), or 12-O tetradecanoylphorbol-13-acetate (TPA) were immunostained for the differentiation markers, human neutrophil elastase (HNE), cathepsin B, MAC387, or the receptor for hepatocyte growth factor (cMET).Results HGF treated cells were positive on staining for cathepsin B and MAC387, but were negative for HNE, indicating monocytic differentiation. HGF treated cells had the morphology of monocytes but continued to divide at the same rate as control cells and remained non-adherent. DMSO treated cells were positive for HNE and cell numbers were reduced, confirming myeloid differentiation. TPA treated cells were positive for cathepsin B and MAC387, cell numbers were reduced, and the cells became adherent, confirming terminal monocytic differentiation. Untreated HL60 cells were weakly positive for cMET at the start of the culture period and expression increased after 72 hours. Cells treated with HGF, DMSO, or TPA were also positive for cMET.Conclusions-These data suggest that HGF induced partial monocytic differentiation in HL60 cells. In addition, expression of cMET by HL60 cells occurs at an early stage in myelomonocytic cells and is maintained after differentiation along either the myeloid or monocytic pathways. PMID- 16695972 TI - Neutrophil elastase and cathepsin G protein and messenger RNA expression in bone marrow from a patient with Chediak-Higashi syndrome. AB - Aims-To determine whether neutrophil elastase and cathepsin G are expressed, at transcriptional or translational levels, in the bone marrow from a patient with Chediak-Higashi syndrome.Methods-Blood neutrophils were isolated from three patients with Chediak-Higashi disease and bone marrow was collected from one. Cell lysates were analysed for neutrophil elastase and cathepsin G activity by enzyme linked immunosorbent assay and western immunoblotting. Northern blotting was used to detect messenger RNA (mRNA) for cathepsin G, elastase and beta-actin in bone marrow extracts, and immunohistochemistry was used to localise the enzymes in marrow myeloid cells.Results-Elastase and cathepsin G were not detected in blood neutrophils from the patients with Chediak-Higashi disease, but were present in bone marrow cells, although immunohistochemistry showed they were not within cytoplasmic granules. The concentrations of elastase and cathepsin G in Chediak-Higashi bone marrow were about 25 and 15%, respectively, of those in normal marrow. Quantitative scanning of northern blots showed that elastase and cathepsin G mRNA, corrected for beta-actin mRNA, were expressed equally in normal marrow.Conclusions-Transcription of elastase and cathepsin G mRNA in promyelocytes of patients with Chediak-Higashi disease is normal, but the protein products are deficient in these cells and absent in mature neutrophils. This suggests that the translated proteins are not packaged into azurophil granules but are degaded or secreted from the cells. PMID- 16695973 TI - Sites of M-CSF messenger RNA production in bone marrow trephine biopsy specimens and long term cultures demonstrated by nonisotopic in situ hybridisation. AB - Aim-To develop methods of messenger RNA (mRNA) in situ hybridisation (ISH) for use with routinely processed bone marrow trephine biopsy specimens, decalcified using formic acid, and long term cultures in order to demonstrate sites of synthesis of mRNA encoding monocyte colony stimulating factor (M-CSF).Methods Biotinylated oligonucleotide probes, directed against target sequences within M CSF mRNA, were hybridised with sections from bone marrow trephine biopsy specimens and detected using Streptavidin-biotin alkaline phosphatase complex formation. Validation of results included demonstration of total mRNA and unrelated mRNA species in adjacent sections, with appropriate negative controls. Minor technical modifications were required to perform ISH with long term bone marrow cultures.Results-M-CSF mRNA was demonstrated successfully in trephine biopsy specimens and long term cultures. Biopsy specimens varied in their requirement for predigestion with proteinase K and in the strength of the final reaction product, presumably due to variation in fixation. M-CSF mRNA was present in myelocytes and promonocytes. No stromal production of M-CSF mRNA was detected in biopsy specimens. ISH using long term bone marrow cultures confirmed production of M-CSF mRNA by developing monocytes and macrophages. Weak M-CSF mRNA expression was also seen in stromal fibroblasts.Conclusions-ISH can be performed successfully with formic acid decalcified bone marrow trephine biopsy specimens and long term cultures. The presence of M-CSF mRNA in myelomonocytic cells suggests that an autocrine mechanism contributes to monocyte differentiation. The absence of detectable M-CSF mRNA in biopsy stroma and its presence in stromal fibroblasts within bone marrow cultures probably reflects reduced sensitivity of ISH following tissue fixation and processing. PMID- 16695974 TI - Activation by interferon-gamma of expression of ICAM-1 and MHC class II antigens in tumour cells from colorectal carcinomas. AB - Aims-To determine whether lack of MHC class II antigen and intercellular adhesion molecule-1 (ICAM-1) expression in some tumours is due to the inability of the tumour cells to respond to the cytokine interferon-gamma (IFN-gamma), an important activator of these surface molecules.Methods-Cells from 40 colorectal tumours which did not constitutively express class II MHC antigens or ICAM-1 were kept in short term culture after disaggregation for a few days to two weeks without significant loss of viability. These were treated with IFN-gamma. Expression of class II MHC antigens and ICAM-1 was determined using immunohistological techniques.Results-There was clear induction in vitro of both MHC class II antigens and ICAM-1 in cells from eight of the tumours, with between 50 and 80% of the tumour cells in the cultures staining positively. The staining was apparent within 24 hours, appeared maximal at about three days, and declined thereafter. There were no obvious differences in cell morphology or viability between the cultures which were inducible and those which were not, nor were there obvious differences between the tumours from which they were derived.Conclusions-Expression of MHC class II antigens and ICAM-1 may be induced by IFN-gamma in a small proportion of colorectal tumours which do not constitutively express these antigens, showing that only a minority of tumours are capable of responding to this cytokine. PMID- 16695975 TI - Peripheral CD25 positive T lymphocytes with biased T cell receptor Vbeta gene usage in autoimmune endogenous posterior uveitis. AB - Aims-To determine T cell receptor (TCR) Vbeta gene usage in peripheral blood T lymphocytes of patients with endogenous posterior uveitis (EPU). If biased TCR variable (V) gene usage occurs in this autoimmune disease, it should be detectable in immune activated peripheral blood T cells in vivo.Methods-Relative proportions of each Vbeta gene family expressed in total peripheral blood lymphocytes (PBL) and in vivo activated (CD25+) T cells from patients with EPU and controls were determined using the anchored polymerase chain reaction (anchored PCR) in conjunction with a novel hybridisation assay. The TCR Vbeta repertoires seen in these cell populations were then compared.Results-Vbeta1 usage within the CD25+ lymphocytes of patients with EPU was substantially elevated (mean +/- SD 15 +/-9%) compared with control CD25+ cells (3.3 +/ 2.4%).Conclusions-By contrasting the repertoires of these cell populations, biased TCR Vbeta gene usage was detected in patients with EPU, namely increased usage of Vbeta1 in CD25+ T cells from peripheral blood of these patients. This approach of directly analysing the activated T cells in blood, using bulk PBL as an internal control, has wide applicability where specific T cell subpopulations are thought to play an important aetiopathological role. PMID- 16695976 TI - Neurological disease associated with Mycoplasma pneumoniae infection. PCR evidence against a direct invasive mechanism. AB - Aims-To investigate the pathology in patients presenting with sudden onset neurological illnesses associated with Mycoplasma pneumoniae infection.Methods-M pneumoniae infection was diagnosed by a highly rigorous interpretation of serological markers initially using complement fixation, agglutination and IgM antibodies. Confirmation of the serological diagnosis was achieved using indirect immunofluorescence for IgM, IgA, and IgG. Serum and cerebrospinal fluid (CSF) samples from these patients were examined using the polymerase chain reaction to look for evidence of M pneumoniae DNA.Results-No M pneumoniae DNA was found in any serum or CSF samples. Diagnosis of M pneumoniae infection by agglutination and complement fixation antibodies was not always confirmed by indirect immunofluorescence.Conclusion-The neurological lesions in these patients do not appear to be caused by the direct invasion of M pneumoniae into the nervous system. The lesions may be an immune response to infection. Serological diagnosis of M pneumoniae continues to be a laboratory problem. PMID- 16695977 TI - Isolation of DNA from archival Papanicolaou stained cytological smears using a simple salting-out procedure. AB - DNA from archival Papanicolaou stained and unstained cytological smears was successfully isolated using a simple, rapid and inexpensive salting-out procedure. The quality of DNA was controlled by polymerase chain reaction (PCR) amplification of segments of the human beta-globin, human beta-actin and human papillomavirus L1 genes. Only negligible differences in amplification efficiency were observed between DNA isolated from stained and unstained smears. The salting out procedure is a more rapid method for the isolation of DNA than phenol chloroform extraction and may be used in instances where fresh or cryopreserved clinical specimens are not available. PMID- 16695978 TI - Clinical gene analysis and manipulation: overview and applications in pathology. PMID- 16695979 TI - A rapid and reliable PCR method for detecting clonal T cell populations. AB - Aims-To establish a reverse transcription polymerase chain reaction (RT-PCR) for the detection of clonal T cell populations, and to evaluate the sensitivity and reliability of the technique.Methods-After reverse transcription of the target RNA with a consensus T cell receptor (TCR) beta constant (C) region primer, consensus C, variable (V), diversity (D) and joining (J) region primers were used to amplify across various portions of the TCRbeta V-D-J-C junction.Results-In normal T cells the polyclonal rearrangements produce a ladder of PCR bands representing the different sized junction fragments. The presence of a T cell clone leads to over-representation of one junction fragment, hence a disproportionately brighter band in the PCR ladder. In a series of 16 patients the RT-PCR detected nine of nine shown to have a clonal TCRbeta rearrangement by Southern blotting and for six of seven patients, it confirmed the presence of a clone indicated by histology or immunophenotyping with FACS analysis, but which was undetectable (five patients) or not investigated (two patients) by Southern blotting. Investigations mixing RNA from normal lymphocytes and the Jurkat TCR Vbeta8 T cell line suggested that the method was more sensitive than Southern blotting.Conclusions-All PCR methods are faster and easier than Southern blotting, but RT-PCR also improves detection of clonal T cell populations, is reliable and produces results that are easy to interpret. PMID- 16695980 TI - Expression of B7 (CD80) and CD40 antigens and the CD40 ligand in Hodgkin's disease is independent of latent Epstein-Barr virus infection. AB - Aim-To examine the expression of CD40 and B7 (CD80) antigens and the CD40 ligand in Hodgkin's disease.Methods-Antigen and ligand expression was studied in 17 cases of Hodgkin's disease using immunohistochemistry. The study included 11 cases of Hodgkin's disease in which latent Epstein-Barr virus (EBV) infection could be demonstrated within tumour cells by in situ hybridisation for the EBV encoded early RNAs (EBERs).Results-In all cases, irrespective of EBV status, Reed Sternberg cells and their variants (HRS cells) showed strong expression of both B7 and CD40 antigens. CD40 ligand expression was not shown in HRS cells but was confined to a subset of small lymphocytes some of which were seen to be in intimate contact with HRS cells.Paraffin wax sections from a further 60 cases of Hodgkin's disease were examined for CD40 and EBER expression alone. The CD40 antigen was identified in HRS cells in all of these cases irrespective of EBER expression.Conclusions-As CD40 and B7 expression are features of professional antigen presenting cells, these results provide further evidence that HRS cells may have antigen presenting properties and that this may contribute to the characteristic recruitment and activation of non-malignant lymphocytes which is a feature of Hodgkin's disease. The ability of HRS cells to activate T(h) cells may in turn contribute to their own survival through the induction of the gp39/CD40 pathway. PMID- 16695981 TI - Absence of Epstein-Barr virus in testicular germ cell tumours: a study of 21 cases using in situ hybridisation. AB - Aim-To establish whether testicular germ cell tumours contain Epstein-Barr virus (EBV) and if so to provide further evidence for the hypothesis that EBV plays a direct role in the pathogenesis of testicular germ cell tumours.Method-Paraffin wax embedded tissue blocks from 21 germ cell tumours including 12 teratomas and nine classic seminomas were examined by in situ hybridisation for the expression of the small EBV encoded nuclear RNA transcripts (EBER-1 and -2) using isotopic and nonisotopic probes.Results-There was no EBER specific signal detectable in any of the testicular germ cell tumours examined by in situ hybridisation whilst a strong signal was observed in appropriate control sections.Conclusion-The absence of demonstrable EBER transcripts in testicular germ cell tumours make a direct role for EBV in the pathogenesis of these tumours unlikely. Other explanations for the epidemiological and serological evidence linking EBV with germ cell tumours need to be explored. PMID- 16695982 TI - Genetic susceptibility to multiple sclerosis in the Shanghai Chinese is not linked to the myelin basic protein gene microsatellite. AB - Aim-To investigate the role of myelin basic protein (MBP) gene polymorphisms in determining susceptibility to multiple sclerosis in a Shanghai Chinese population.Methods-Forty seven unrelated patients with multiple sclerosis and 94 healthy control subjects were included in the study. Genomic DNA was extracted from peripheral blood lymphocytes and amplified using the polymerase chain reaction to characterise two adjacent tetranucleotide repeats ([ATGG](12) and [TGGA](9)) located 5' to exon 1 of the MBP gene.Results-Two polymorphic loci were identified: locus A, comprising both repeats, and locus B, comprising the [ATGG](12) repeat only. Nine allelic variants were identified at locus A and six at locus B, ranging from 212 to 244 and 122 to 146 base pairs, respectively. The 244 base pair allele at locus A has not been reported before. The allele frequencies observed in the controls differed from those seen in normal white populations.Conclusions-The present study demonstrates a race specific pattern of allelic distribution within the tetranucleotide repeat of the MBP gene. Further studies are needed to fully elucidate the role of the MBP gene in inherited susceptibility to multiple sclerosis. PMID- 16695983 TI - Protein kinase C: a family of isoenzymes with distinct roles in pathogenesis. PMID- 16695985 TI - Loss of heterozygosity in lobular carcinoma in situ of the breast. AB - Aims-(1) To investigate whether loss of heterozygosity identified at various loci in invasive breast carcinoma or is present in lobular carcinoma in situ (LCIS). (2) To investigate whether LCIS is a monoclonal (neoplastic) or a polyclonal (hyperplastic) proliferation.Methods-Forty three cases of LCIS (30 with associated invasive carcinoma or in situ ductal carcinoma (DCIS) and 13 cases of pure LCIS) were investigated for loss of heterozygosity on chromosomes 16q, 17q, 17p, and 13q using a microdissection technique, polymorphic DNA markers, and the polymerase chain reaction (PCR).Results-Loss of heterozygosity was detected in both subgroups of LCIS at all the loci examined. There was no significant difference in the frequency of the loss between the group associated with invasive carcinoma and the pure LCIS group. The frequency of loss of heterozygosity ranged from 8% on 17p to 50% on 17q.Conclusions-Because of the nature of the technique employed, our findings show that LCIS is a monoclonal (neoplastic) proliferation rather than a hyperplastic proliferation. The incidence of loss of heterozygosity on 17p (D17S796) is lower than we have observed previously in DCIS, suggesting that LCIS and DCIS are different genetically as well as clinically and morphologically. The similar incidence of loss of heterozygosity on 16q and 17q, however, suggests that DCIS and LCIS may share a common pathway of evolution. PMID- 16695984 TI - The study of minimal residual disease in acute lymphoblastic leukaemia. PMID- 16695986 TI - p53 overexpression in Ewing's sarcoma/primitive neuroectodermal tumour is an uncommon event. AB - Aim-To determine the presence of p53 overexpression in Ewing's sarcoma/primitive neuroectodermal tumours (ETs) and to assess whether p53 accumulation has any prognostic value.Methods-From a prospectively compiled database of 76 patients with ETs, suitable tumour tissue was available for 38. The monoclonal antibody pAb1801 was used to detect p53 nuclear protein overexpression. Results-Nuclear staining was detected in the tumours of three (8%) of the 38 patients. Where tumours stained positively, over 10% of the tumour nuclei were postively stained. All three patients whose tumours overexpressed p53 died and in a relatively short time compared with the patients who did not overexpress p53 (mean 3.7 months compared with a mean of 38.7 months in the p53 negative group).Conclusion Overexpression of p53 in ETs is an uncommon event. Overexpression of p53 has repeatedly been shown to correlate closely with p53 point missense mutations and therefore this oncogenic event appears not to be of primary pathogenic importance in ETs. There is a tentative indication that those uncommon ETs in which p53 overexpression can be detected may be-have more aggressively. PMID- 16695988 TI - Stathmin is expressed by the proliferating hepatocytes during liver regeneration. AB - Aim-To determine the liver cell populations that express the phylogenetically conserved cytosolic protein stathmin during liver regeneration.Methods-Double immunostaining for stathmin and the Ki67 antigen was performed on sections of formaldehyde fixed, paraffin wax embedded tissues from 31 liver specimens. These included a variety of disease conditions characterised by some degree of hepatocyte regeneration. Quantitative western blot analysis was performed on 22 of these specimens.Results-Variable amounts of stathmin protein were detected by western blotting in all of the specimens examined. Stathmin was not detected in three cases of histologically normal liver. On immunostaining, stathmin was demonstrated in a proportion of hepatocytes as well as lymphoid inflammatory cells and other tissue elements. In all cases most of these stathmin positive cells showed nuclear positivity for the Ki67 antigen.Conclusions-Stathmin is expressed by proliferating hepatocytes but not by resting hepatocytes. Thus, it is likely that the protein has a function important to cell proliferation as opposed to cell differentiation. PMID- 16695987 TI - A comparative study of cell proliferation markers in breast carcinomas. AB - Aims-To investigate the tumour cell proliferative index obtained by immunostaining of paraffin wax sections of 30 cases of breast carcinoma with monoclonal antibodies MIB1, KiS1 and KiS5, and polyclonal Ki67 antisera to the Ki67 antigen and 19A2 and PC10 antibodies to proliferating cell nuclear antigen and the possible correlation between these indices and that of monoclonal Ki67 antibody in frozen sections of the same tumours.Methods-All tumour samples had been uniformly fixed and processed and sections were subjected to microwave antigen retrieval before immunostaining in all instances except for monoclonal Ki67 antibody which was used in cryostat sections. Tumour cell proliferative indices were evaluated by two independent examiners, each counting 500 tumour cells with the aid of a cross-hatched grid.Results-Proliferative indices obtained with MIB1, polyclonal Ki67, KiS1, and KiS5 correlated with those obtained with monoclonal Ki67 in frozen sections. Proliferative indices obtained with monoclonal 19A2 and PC10 showed no correlation with those of monoclonal Ki67 antibody. The staining obtained with MIB1 was the most intense and the easiest to read.Conclusions-Monoclonal antibodies MIB1, KiS1 and KiS5 and polyclonal Ki67 antiserum appear to be suitable substitutes for monoclonal antibody Ki67 in the assessment of tumour cell proliferative index. As these reagents are all immunoreactive in paraffin wax sections, they overcome the requirement for frozen tissue for immunostaining with monoclonal Ki67. PMID- 16695989 TI - Differential expression of cell adhesion molecules in the functional compartments of lymph nodes and tonsils. AB - Aims-To analyse the topographical distribution of adhesion molecules involved in lymphocyte recirculation in human lymph nodes and tonsils. The study focused on the expression of LECAM-1 (CD62L), VLA-alpha4 (CD49d), VLA-beta1 (CD29), LFA-1 alphaL (CD11a), LFA-beta2 (CD18), VCAM-1 (CD106), ICAM-1 (CD54), and H-CAM (CD44).Methods-Reactive lymph nodes and palatine tonsils were studied using immunofluorescence methods with fluorescein isothiocyanate (FITC) labelled monoclonal antibodies directed against cell adhesion molecules. To investigate the expression patterns of these molecules in the T and B cell populations, double labelling experiments were performed using Texas Red labelled antibodies against CD2 or CD19, respectively. The images from each fluorochrome were then simultaneously analysed using a laser scanning confocal microscope.Results-LECAM 1, VLA-alpha4 and H-CAM were predominantly expressed by mantle zone B cells, VCAM 1 and ICAM-1 by germinal centre cells, most of which exhibited a reticular staining pattern suggestive of follicular dendritic cells, whereas LFA-1 alphaL and LFA-beta2 were mainly found in extrafollicular and germinal centre T cells. All high endothelial venules expressed VLA-beta1 and ICAM-1, whereas VCAM-1 was present in only a few, with variable intensity.Conclusions-The data show that all of these adhesion molecules are differentially distributed within the distinct functional microenvironments of both organs. The differences observed in the expression patterns among the B and T cells belonging to different compartments probably depend on the momentum of cell traffic, the stage of maturation/activation, as well as on their functional role in the immune response. PMID- 16695990 TI - Ki67: from antibody to molecule to understanding? PMID- 16695991 TI - Does junk DNA regulate gene expression in humans? PMID- 16695992 TI - Rapid identification of mycobacteria from AIDS patients by capillary electrophoretic profiling of amplified SOD gene. AB - Aim-Rapid differentiation of mycobacterial species at the genomic level.Methods The manganese superoxide dismutase (SOD) gene (464 bp) and 16SrRNA (353 bp) from 104 isolates (18 species) of mycobacteria were amplified using polymerase chain reaction (PCR). Products were sequenced and a phenogram of SOD sequences derived. PCR products of SOD gene were digested with HaeIII, and restriction fragment profiles visualised using capillary electrophoresis.Results-Novel SOD sequences were found for M szulgai, M marinum, M phlei, M smegmatis, M chelonei, M paratuberculosis, M malmoense, M intracellulare serotype 7, M intracellulare serotype 18, and M celatum types 1, 2, and 3. Phylogenetic analysis indicated that 18 of 19 species studied had 8-29% interspecies and <6% intraspecies sequence diversity in the SOD gene. No consistent differences were detected between AIDS and non-AIDS isolates. M paratuberculosis showed a unique SOD sequence with a 1.1% (SD 0.5%) diversity from M avium. Capillary electrophoresis profiles were able to differentiate 16 of 18 species within 24 hours.Conclusions A phenogram of SOD sequences clearly delineated all mycobacterial species and showed two distinct clusters, fast growing species, and the M avium complex (MAC). Within the MAC, M avium (five types), M intracellulare (five types), M scrofulaceum (two types), and M paratuberculosis (one type) could be demonstrated. Phylogenetic diversity of M celatum from MAC, previously suggested by 16SrRNA data, was confirmed. This simple and rapid method for DNA extraction, in conjunction with capillary electrophoresis of SOD restriction fragments, allows rapid identification of mycobacterial isolates. PMID- 16695993 TI - Comparison of random amplified polymorphic DNA with restriction fragment length polymorphism as epidemiological typing methods for Mycobacterium tuberculosis. AB - Aim-To compare restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) methods for the epidemiological typing of Mycobacterium tuberculosis.Methods-Thirty one M tuberculosis cultures originating from patients in the Canton of Berne in Switzerland, which had previously been typed by RFLP, were subjected to RAPD analysis. Cultures were coded so that the investigators were blind to the RFLP results until RAPD analysis was complete.Results-The 31 cultures of M tuberculosis were divided into nine groups by RFLP and eight groups by RAPD. Generally there was good correlation between the groups identified by the two techniques, with the exception of strains that had only one copy of IS6110. Both methods subdivided isolates that were placed in a single group by the other method.Conclusions-RAPD analysis is quick, simple, and useful for the comparison of small numbers of isolates. RFLP is more reproducible and therefore better suited for the accumulation of RFLP fingerprints for long term local surveillance and large epidemiological studies. PMID- 16695994 TI - Selection of peptide ligands for the antimucin core antibody C595 using phage display technology: definition of candidate epitopes for a cancer vaccine. AB - Aims-To further define the specificity of the antimucin core antibody C595 by fitting it with a family of hexapeptide ligands by immunoselection of filamentous bacteriophage from a gene III display library of approximately 6.4 x 10(7) random hexapeptides.Methods-Three rounds of immuno-selection were used to enrich for C595 binding phage. DNA sequencing revealed the hexapeptides expressed. Bacteriophage and corresponding synthetic hexapeptides were used in ELISA assay to determine binding affinities.Results-Twenty nine clones from this selected population were analysed. Seven contained the natural epitope RPAP, encoded by two different DNA sequences; 17/29 contained the motif RLPP. In all, 28/29 clones contained the motif RXXP and one clone (RVRPAP) contained the motif RXXP in two peptidic registers; 24/28 clones (6/8 DNA sequences) contained a hydrophobic residue (V or I) at position 1 relative to the RXXP motif. In addition the proximity of RXXP to glycine (position 5) suggests that this contributes in the natural epitope to antibody/antigen binding, which was not detected by chemical synthetic methods. One clone, KSKAGV, bears no obvious relationship to the natural epitope and therefore qualifies as a weakly binding mimotope.Conclusions This approach has rapidly defined the specificity of this antibody in unprecedented detail, and provides a more comprehensive molecular basis for exploring the immune recognition of the MUC1 mucin by the C595 antibody. Importantly, the novel but related epitopes seen provide peptide specificities and a strategy which may prove useful in generating cancer vaccine candidates. PMID- 16695995 TI - Non-invasive detection of bladder cancer by identification of abnormal CD44 proteins in exfoliated cancer cells in urine. AB - Aims-To investigate the expression of CD44 proteins in exfoliated urothelial cells and in tumour tissues from bladder cancer patients. A further objective was to evaluate the diagnostic potential of the changes observed in the expression of these proteins as a marker for non-invasive detection of bladder cancer.Methods Naturally voided urine specimens were collected from 47 patients with bladder cancer or severe urothelial dysplasia (n=3) and from a control group of 43 people with no evidence of neoplastic disease. Exfoliated urothelial cells floating in the urine were pelleted by centrifugation and lysed, and their constituent proteins extracted. The pattern of expression of CD44 proteins in each sample was examined by western blot analysis using a monoclonal antibody, Hermes 3, which recognises an epitope on the polypeptide backbone of the CD44 protein. Immunohistochemical studies were performed on neoplastic (n=10) and normal (n=4) bladder tissue specimens which were snap frozen in liquid nitrogen before examination with antibodies to CD44 gene products (CD44s and CD44v6).Results Western blot analysis revealed several high molecular weight CD44 isoforms > 160 kDa in urine cell lysates from 75% of patients with histologically confirmed bladder cancer and in two of the three patients with severe dysplasia. Such patterns were not detected in the urine cell pellets from any persons in the control group. Immunohistochemical studies of the tissue distribution of CD44s and CD44v6 showed that the differentiation and maturation of the epithelial cells in the normal bladder mucosa is accompanied by a decrease in CD44 protein expression. However, carcinoma cells overexpress standard and variant CD44 isoforms and continue to do so as they proceed through the thickened epithelial layer to the luminal surface and after they are shed into the urine.Conclusions The abnormal expression of CD44 proteins in exfoliated cancer cells may be a useful marker for the noninvasive diagnosis of bladder cancer. PMID- 16695996 TI - Circulating human IgG autoanti-IgE antibodies in asthma patients block the binding of IgE to its high affinity receptor. AB - Aims-To investigate the ability of circulating human IgG autoanti-IgE antibodies from asthma patients to block the binding of IgE to the alpha chain of the high affinity receptor (FcepsilonRI).Methods-This involved the use of a well validated flow cytometric method to detect inhibition of FITC labelled IgE binding to a fibroblast cell line (CHK1E1) transfected with the alpha chain of FcepsilonRI.Results-IgG autoanti-IgE-containing sera blocked the binding of IgE FITC to the CHK1E1 cells. No such inhibition was demonstrable with rheumatoid sera containing autoanti-IgG (that is, rheumatoid factor) but lacking autoanti IgE. Percentage inhibition (up to 50%) of IgE binding to the CHK1E1 cells was directly related to the titre of IgG1, but not IgG4, autoanti-IgE in the sera tested (correlation coefficient 0.66, probability 0.003).Conclusions-The capacity of anti-IgE to block the binding of IgE to FcepsilonRI has important clinical implications, particularly in terms of downregulation of allergic reactions. PMID- 16695997 TI - Insulin-like growth factor II prevents apoptosis in a human teratoma derived cell line. AB - Aim-To study how insulin-like growth factor II (IGF-II) affects the behaviour of human teratoma cells.Methods-The human pluripotential teratoma cell line Tera 2 was cultured under serum-free conditions in the presence or absence of IGF-II. Effects on cell proliferation and apoptosis as well as on the expression of the proto-oncogene c-myc were studied.Results-In this study we show that Tera 2 cells deprived of serum undergo programmed cell death (apoptosis). The onset of nuclear fragmentation was observed 12 hours after serum withdrawal. The morphological changes of the Tera 2 cell nuclei were confirmed by the occurrence of a nucleosome ladder. However, the constitutive expression of the proto-oncogene c myc was not decreased in parallel with initiation of apoptosis. The apoptotic response to serum withdrawal could be counteracted by simultaneous addition of IGF-II. In addition it was found that human testicular tumours (seminoma and embryonal carcinoma) contain raised levels of insulin-like growth factors.Conclusions-The precise roles of IGF-I and IGF-II have been unclear, and there is overwhelming evidence against these factors as primarily transforming agents. The finding that IGF-II apparently counteracts apoptosis in vitro may well explain its effects on tumours in vivo. PMID- 16695998 TI - In situ visualisation of immunoglobulin genes in normal and malignant lymphoid cells. AB - Aims-To directly visualise immunoglobulin (Ig) heavy (H) and light chain genes (kappa and lambda) in metaphase chromosomes and interphase nuclei of normal and malignant lymphocytes using small genomic probes targeted to intragenic sequences.Methods-Cytogenetic preparations from phytohaemagglutinin stimulated lymphocytes, B-chronic lymphocytic leukaemia (B-CLL) cells, and a B prolymphocytic leukaemia (B-PLL) cell line, containing a t(11;14), were hybridised in situ using biotin or digoxigenin labelled plasmid probes. The kappa genes were visualised with a combination of probes for the Ckappa, Jkappa, Vkappa1, and Vkappa2 segments, the lambda genes with a probe containing the Jlambda2-Clambda2, Jlambda3-Clambda3 segments and the H genes with a probe for Clambda2. Hybridisation sites were visualised using appropriate fluorochrome conjugates and images were analysed by digital microscopy.Results-In both normal and malignant lymphoid cells, the kappa and lambda genes were visualised as a single dot signal, whereas the H lambda genes were resolved as either two or three separate signals per chromatid in metaphase chromosomes or per allele in interphase nuclei. In the malignant PLL cells, double hybridisation experiments with a painting library specific for the chromosome 11 showed that the lambda region was retained in the translocated chromosome, with an in situ resolution pattern similar to that of the normal allele.Conclusions-This study shows that a high resolution in situ analysis of the three Ig loci can be efficiently performed with small size genomic probes on both normal and malignant lymphoid cells. Such an approach offers a flexible tool for the molecular characterisations of these loci on chromosomes and individual neoplastic cells. PMID- 16695999 TI - Leukaemic mantle cell lymphoma with t(11;14) and trisomy 12 showing clinical features of state A0 B cell chronic lymphocytic leukaemia. AB - The precise diagnosis of lymphoma usually requires the histological examination of lymph nodes or involved tissues. Mantle cell lymphoma is a form of intermediate grade non-Hodgkin's lymphoma in which typical morphological immunophenotypic and cytogenetic features have been recognised. A case of leukaemic mantle cell lymphoma with the characteristic reciprocal translocation t(11;14) together with trisomy 12, a chromosomal abnormality usually associated with B cell chronic lymphocytic leukaemia (CLL), is presented. This combination of cytogenetic abnormalities has not been reported previously. The lack of lymphadenopathy and hepatosplenomegaly in this patient is more in keeping with stage A0 CLL. This case demonstrates the close clinical and biological relationship between mantle cell lymphoma and CLL. PMID- 16696000 TI - Proteolytic mechanisms of cartilage breakdown: a target for arthritis therapy? PMID- 16696001 TI - The scope of quantitative polymerase chain reaction assays in clinical molecular pathology. PMID- 16696002 TI - Deletions in the Epstein-Barr virus latent membrane protein-1 oncogene in Hodgkin's disease. AB - Aims-To analyse the latent membrane protein-1 (LMP-1) gene in a series of patients with Epstein-Barr virus (EBV) positive LMP expressing ordinary and HIV associated Hodgkin's disease to detect possible genetic alterations and particularly the existence of deletions near the 3' end of the gene.Methods Expression of the EBV LMP-1 was assessed using immunohistochemistry in 186 cases of Hodgkin's disease and 31 cases of HIV associated Hodgkin's disease. Genomic DNA was extracted from frozen lymph node biopsy specimens from 25 cases of Hodgkin's disease and 11 of HIV associated Hodgkin's disease, all of whom expressed the LMP-1 protein within diagnostic Hodgkin and Reed-Sternberg (HRS) cells, and amplified by polymerase chain reaction (PCR) using primers specific for the different LMP-1 regions.Results-LMP-1 expression was observed in 106 of 186 Hodgkin's disease cases and in all 31 HIV associated Hodgkin's disease cases. Molecular analysis of the LMP-1 gene showed a high degree of genetic heterogeneity in the carboxy-terminal domain compared with the prototype B95-8 EBV strain, specially in the patients with HIV associated Hodgkin's disease. Variation in the size of the repeated region was found in 17 of 25 Hodgkin's disease and nine of 11 HIV associated Hodgkin's disease cases. Deletions of 30 base pairs near the 3' end of the gene were detected in all cases of HIV associated Hodgkin's disease and in six Hodgkin's disease. In one case of Hodgkin's disease a larger deletion was observed. In all patients with LMP-1 deletion mutants, 50-90% of the diagnostic HRS cells expressed the LMP-1 protein.Conclusions-The presence of the 30 base pair deletion in all cases of HIV associated Hodgkin's disease supports previous studies that reported aggressive histological and clinical behaviour in tumours harbouring this deletion. This deletion may prolong the half-life of the protein which would explain the high levels of LMP-1 expressing HRS cells in those cases carrying LMP-1 deletions. That the 30 base pair deletion was present in all of the HIV associated Hodgkin's disease specimens suggests that impairment of immune function is a stringent requirement for the expansion of malignant cells infected by EBV strains containing the deleted LMP-1 gene. PMID- 16696003 TI - Absence of Epstein-Barr virus in oesophageal squamous cell carcinoma. AB - Aim-To identify the possible role of Epstein-Barr virus (EBV), in Chinese patients, in the pathogenesis of oesophageal squamous cell carcinoma.Methods Formalin fixed, paraffin wax embedded tissues from 74 cases of oesophageal squamous cell carcinoma (28 with well differentiated, 27 with moderately differentiated and 18 with poorly differentiated carcinomas) were analysed for EBV using in situ hybridisation for EBV encoded small RNAs.Results-EBV was detected in only a few lymphocytes adjacent to the tumour epithelia in 14 (19%) cases of oesophageal carcinoma. The adjacent, non-pathological oesophageal tissue was EBV negative.Conclusions-EBV does not play a major role in the aetiology of oesophageal squamous cell carcinoma. PMID- 16696004 TI - Comparison of different Ki67 antibodies in human glioblastomas. AB - Aims-To compare immunostaining between the original Ki67 monoclonal antibody and a new polyclonal Ki67 antibody on frozen and paraffin wax sections of human glioblastomas.Methods-Frozen sections and formalin fixed, paraffin wax embedded sections of the same tumour specimens were included in the study (10 cases). Half of the paraffin wax sections were pretreated in a microwave oven. Standard immunohistochemical techniques were used (avidinbiotin peroxidase complex). Five high power fields were examined using an eye-piece graticule, and 500 to 2000 tumour cells were counted. The labelling index was defined as the percentage of positive tumour cells.Results-The Ki67 monoclonal antibody displayed positive immunostaining in all frozen sections (median labelling index 5.9, range 2.6 11.4) whereas only four paraffin wax sections stained positively and only after pretreatment in a microwave oven. The polyclonal Ki67 antibody elicited positive staining in both frozen sections (median labelling index 13.7, range 6.7-21.5) and in paraffin wax sections (median labelling index 12.0, range 2.2-22.7) but only after pretreatment in a microwave oven.Conclusion-The Ki67 monoclonal antibody is not recommended for use on paraffin wax sections of glioma tissues whereas the new polyclonal Ki67 antibody provides satisfactory immunostaining on both frozen and paraffin wax sections. PMID- 16696005 TI - p53 Overexpression in laryngeal squamous cell carcinoma and dysplasia. AB - Aim-To investigate the expression of p53 protein in invasive squamous cell carcinoma (SCC) of the larynx and dysplasia in relation to histological grade and tobacco smoking.Method-Paraffin wax embedded tissue sections from 41 cases of invasive SCC of the larynx, 28 cases of dysplasia and 14 control laryngeal biopsy specimens were studied immunohistochemically using two anti-p53 monoclonal antibodies (DO7 and 1801). The Streptavidin/horseradish peroxidase method was used after microwave antigen retrieval and a semiquantitative method was applied to assess the extent of p53 expression.Results-Of the cases of invasive SCC of the larynx, 78% (32/41), regardless of histological grade, overexpressed p53 compared with only 30% (eight of 28) of cases of mild dysplasia. A gradual increase in p53 expression from mild to severe dysplasia (60%) was observed, and only three of 14 control biopsy specimens of laryngeal nodules showed occasional weakly positive basal cells.Conclusion-The gradual increase in p53 expression from mild to severe dysplasia to invasive SCC indicates that p53 overexpression is an early event in laryngeal carcinogenesis which may lead to invasive malignancy. p53 overexpression may be related to environmental factors as most of the patients smoked tobacco. Microwave postfixation may be essential for the reliable detection of p53. PMID- 16696006 TI - Absence of c-Ki-ras gene mutation in malignant and premalignant Barrett's oesophagus. AB - Aims-To establish the prevalence of c-Ki-ras gene mutations in codons 12 and 13 in 28 surgically resected Barrett's adenocarcinomas and 18 associated preneoplastic lesions in Barrett's oesophagus.Methods-Mutations were detected using the polymerase chain reaction followed by restriction fragment length polymorphism analysis. Human colon carcinoma cell lines with well characterised mutations in codons 12 and 13 were used as positive controls and to test the sensitivity of the method.Results-c-Ki-ras gene mutations were not detected in any of the 28 specimens of Barrett's adenocarcinoma or in the 18 specimens of Barrett's oesophagus (nine non-dysplastic cases, three cases with low and six with high grade dysplasia).Conclusions-These results suggest that the c-Ki-ras gene is not involved in the development of cancer in Barrett's oesophagus. PMID- 16696007 TI - Prognostic significance of CD44 expression in adenocarcinoma of the lung. AB - Aims-To determine whether expression of CD44 in neoplasia is associated with tumour grade, stage and prognosis.Methods-The immunohistochemical expression of CD44 was evaluated using the mouse antihuman monoclonal antibody 3G12 which recognises regions shared by all CD44 isoforms to determine whether expression in formalin fixed, paraffin wax embedded tissue correlates with tumour grade, stage or survival in adenocarcinoma of the lung. Thirty one adenocarcinomas of the lung, 16 T2N0 and 15 T2N1, and their nodal metastases were studied.Results-Of the 31 tumours, 25 were positive for the CD44 antigen. CD44 expression correlated with tumour grade, in that intense staining was seen only in moderately and/or poorly differentiated tumours. CD44 did not correlate with nodal status, tumour size, pleural invasion, angiolymphatic invasion, or host inflammatory response, but did correlate with survival. A median survival of 46 months was observed in patients with moderate to strong CD44 expression compared with 24 months for those with no or weak expression. Nine patients were alive without evidence of disease at a median follow up of 61 months. Six (66%) of these nine patients had strong CD44 expression. This contrasts with strong expression in only three (17%) of the 17 patients dying with a median survival of 28 months.Conclusion-In primary adenocarcinoma of the lung loss of CD44 expression is associated with less favorable outcome and may indicate a more aggressive neoplasm. CD44 may be a useful prognostic marker in lung carcinoma. PMID- 16696008 TI - Expression of hepatocyte growth factor receptor (c-met) mRNA in primary cultures of human hepatocytes. AB - Aim-To investigate the regulation of hepatocyte growth factor (HGF) receptor (c met) gene expression in isolated primary human hepatocytes.Methods-Primary hepatocytes were maintained in monolayer culture for up to 72 hours in serum-free medium. They were treated with growth factors and the level of HGF, c-met and reduced glyceraldehyde-phosphate dehydrogenase mRNA expression determined by northern blot analysis.Results-Hepatocytes expressed a single 9 kilobase c-met gene transcript whilst HGF mRNA analysis was negative. Addition of HGF and epidermal growth factor, both potent mitogens for human hepatocytes, enhanced c met mRNA expression approximately twofold within 24 hours, after which levels returned to normal. In non-growth factor treated cells, transforming growth factor-beta (TGFbeta) had little effect upon c-met mRNA levels. However, TGFbeta inhibited the HGF induced increase in c-met mRNA levels.Conclusions-These results indicate that hepatocytes which proliferate in response to HGF demonstrate levels of c-met mRNA which are subject to growth factor modulation and suggest an important means of growth regulatory control. PMID- 16696010 TI - Jh gene sequence analysis in a patient with Waldenstrom's macroglobulinaemia with subsequent development of immunoblastic lymphoma. AB - A case of Waldenstrom's macroglobulinaemia with subsequent development of immunoblastic lymphoma in a 69 year old man is reported. Plasmacytoid lymphocytes were initially observed in both peripheral blood and bone marrow smears. Lymph node biopsy was interpreted as malignant lymphoma, diffuse mixed cell type compatible with Waldenstrom's macroglobulinaemia. Complete remission resulted on combined treatment with prednisolone and melphalan. Four years later, lymphadenopathy recurred and biopsy revealed the development of an immunoblastic lymphoma. Initially, this lymphoma was thought to be the malignant transformation of Waldenstrom's microglobulinaemia, because both tumours produced IgM (kappa light chains). Sequencing of immunoglobulin heavy chain genes ofWaldenstrom's macroglobulinaemia and immunoblastic lymphoma cells, however, revealed different Dh and Jh usage between the two, indicating the independent nature of the two haematological malignancies. These results indicate that clonality of primary and secondary tumours should be determined not by phenotypic, but by genotypic analysis. PMID- 16696009 TI - Association between the CYP1A1 gene polymorphism and susceptibility to emphysema and lung cancer. AB - Aim-To investigate cytochrome P4501A1 (CYP1A1) polymorphism and susceptibility to emphysema and lung cancer.Methods-A novel polymerase chain reaction (PCR) for genotyping the CYP1A1 polymorphism, corresponding to putative low or high enzyme activity, was developed to genotype lung cancer resection samples which had been assessed macroscopically for the presence of centriacinar and panacinar emphysema. Samples were collected and genotyped from a group of patients with chronic obstructive airways disease. A control group of anonymous blood donations was genotyped to determine the basal levels of the polymorphism in the Scottish population.Results-The high activity allele of the CYP1A1 gene is associated with susceptibility to centriacinar emphysema and lung cancer but not panacinar emphysema. CYP1A1 polymorphism is not linked to lung cancer in the absence of emphysema, nor to chronic obstructive airways disease which is the clinical manifestation of emphysema, particularly of the panacinar type.Conclusions Susceptibility to emphysema and lung cancer is associated with polymorphism of the P4501A1 gene. A trend towards damage of centriacinar pattern has been detected, which supports the theory that centriacinar emphysema results from local, direct damage to the respiratory bronchioles from exposure to cigarette smoke. PMID- 16696011 TI - Central nervous system involvement by Mycoplasma pneumoniae. PMID- 16696012 TI - Quantitative analysis of silver stained nucleolar organiser regions: a reliable marker of cell proliferation and a promising prognostic parameter in tumour pathology. PMID- 16696014 TI - AgNOR measurements as indices of proliferation, ploidy and prognosis. PMID- 16696015 TI - Isolation and characterisation of antibodies which specifically recognise the peptide encoded by exon 7 (v2) of the human CD44 gene. AB - Aims-Exon 7 of the human CD44 gene is overexpressed in many commonly occurring carcinomas. The aim of the study was to explore the diagnostic and therapeutic potential of this frequent abnormality.Methods-A new monoclonal antibody (mAb, M 23.6.1) and a polyclonal antibody (pAb,S-6127) to the corresponding antigen were raised by immunising mice and sheep, respectively, with a specially constructed fusion protein HIV2 (gp32)-CD44 exon 7.Results-Characterisation of mAb, M-23.6.1 by ELISA, western blotting, immunocytochemistry, and FACS analysis confirmed that it specifically recognises an epitope in the region between amino acids 19 and 33 of the peptide encoded by this exon. Western blotting experiments with two cell lines, RT112 and ZR75-1, known from RT-PCR data to be overtranscribing the exon, yielded a monospecific band of approximately 220 kDa, and immunocytochemistry showed discrete membrane staining on the same cell lines. Fluorescent antibody cell sorting (FACS) revealed binding to greater than 90% of the cells of each of these lines. Specificity of recognition of the antigen was shown by inhibition of the precise immunoreactivity typically seen in ELISA and Western blots, by pre incubation with synthetic exon 7 peptide or fragments of it.Conclusions-The new antibodies will be useful tools for the further analysis of abnormal CD44 isoforms and their clinical implications. PMID- 16696016 TI - Proliferative activity and apoptosis of Langerhans histiocytes in eosinophilic granulomas as evaluated by MIB-1 and TUNEL methods. AB - Aims-To identify the role played by apoptosis in tumour regression.Methods-The growth fraction and apoptotic cell loss of four cases of eosinophilic granuloma were investigated using monoclonal antibodies against Ki-67 proliferation marker (MIB-1) antigen and the TdT mediated dUTP-biotin 3'-OH nick end labelling (TUNEL) method. These data were then compared with the clinical growth rate.Results-Only the Langerhans histiocytic cells, which reacted positively with anti-S-100 protein antibody, were immunolabelled with antibodies to proliferating cell nuclear antigen and Ki-67 antigen (MIB-1). Many apoptotic figures of histiocytic cells were also detected in all cases by the TUNEL method. In a patient whose tumour clinically showed spontaneous regression, the TUNEL staining index gave a higher score than the MIB-1 staining index.Conclusions-The main cause of the spontaneous regression of the tumours was postulated to be programmed cell death (apoptosis). PMID- 16696017 TI - Amplification and overexpression of the cyclin D1 gene in head and neck squamous cell carcinoma. AB - Aims-To determine cyclin D1 gene amplification and expression levels in head and neck squamous cell carcinoma (HNSCC) patients.Methods-Total RNA and genomic DNA were isolated from 40 samples of HNSCC tissue and matched normal tissue and were hybridised with a cyclin D1 cDNA probe. Northern and Southern analyses were used to detect mRNA overexpression and cyclin D1 gene amplification, respectively.Results-15 of the 40 HNSCC samples examined (38%) showed cyclin D1 gene amplification. Of these 15 samples, all 13 from which RNA was available showed increased cyclin D1 expression.Conclusions-HNSCC patients with both amplification and overexpression of the cyclin D1 gene are at greater risk than patients who showed no cyclin D1 gene amplification; amplification and over expression of the cyclin D1 gene may play an important role in the progression of HNSCC and in clinical outcome. PMID- 16696013 TI - Biological indices in the assessment of breast cancer. PMID- 16696018 TI - Interphase ribosomal RNA cistron staining in chronic myeloid leukaemia. AB - Aim-To evaluate the haemopoietic function of bone marrow blood forming cells in human chronic myeloid leukaemia (CML) by means of silver staining of nucleolar organiser region (AgNOR).Methods-Nucleoli were investigated in bone marrow blast cells and in erythroid, granulocytic, and megakaryocytic cells from 10 haematologically healthy subjects and from 26 patients with chronic myeloid leukemia (17 in benign phase, nine with blast crisis). The investigation was done before treatment, by means of a one step silver staining method. In every case 50 to 100 blasts, promyelocytes, myelocytes, immature (pronormoblastic and basophilic normoblastic) and mature (polychromatic normoblastic) erythroid elements, and megakaryocytes were evaluated for the mean numbers of nucleoli and for the average number of AgNORs per nucleus. Student's t test was used to compare the patient and control groups. Other statistical analyses were carried out by means of the computer assisted "HEMA" system.Results-Compared with controls, activation of NORs was noticed only in CML blasts, while there was a decrease in NORs in the erythroid elements, promyelocytes, and megakaryocytes. The AgNOR score of polychromatic normoblasts and megakaryocytes started to decrease in the benign stage of CML, whereas a similar decrease in pronormoblasts, basophilic normoblasts, and promyelocytes was detected only in patients with CML blast crisis.Conclusions-The loss of AgNOR sites in cell series in CML may be related to intrinsic defects in their proliferation. PMID- 16696019 TI - Proportionally constant quantitative transmission of nucleolin and protein B23 in cycling cancer cells. AB - Objective-To investigate whether and to what extent the two major AgNOR proteins, nucleolin and protein B23, are maintained after one cell division in proliferating cells.Design-Using three asynchronously growing human cancer cell lines, TG, SJNKP, and CHP 212 cells, nucleolin and protein B23 were first identified on SDS-polyacrylamide separated nucleolar proteins, transferred to nitrocellulose and silver stained for AgNOR proteins. Measurement of doubling time indicated a period very close to 24h for each of the cell lines. To quantify the percentage of nucleolin and protein B23 maintained in daughter cells after duplication, cells were labelled with [(35)S]-methionine and a 24h cold chase performed. Nucleolin and protein B23 labelling was evaluated by densitometric analysis on nitrocellulose autoradiograms.Results-The radioactivity relative to nucleolin and protein B23 bands maintained in the daughter cells was a constant fraction of that present before cell duplication. In the three cell lines the percentage of residual radioactivity measured in the nucleolin bands was 42.2, 40.6, and 41.2 and in protein B23 bands 48.0, 46.2, and 44.1.Conclusions-After one cell division the nucleolin and protein B23 quantity present in cells may be highly variable, depending on the amount of the two proteins present in the mother cell. This is important in relation to the correct utilisation of AgNOR protein quantity as an index for evaluating cell kinetics. PMID- 16696020 TI - Altered p53 in microdissected, metachronous, premalignant and malignant oral lesions from the same patients. AB - Aims-To determine whether mutant p53 alleles harboured by malignant tumours of the oral cavity were also present in previous premalignant lesions at the same site.Methods-Paraffin embedded tumour specimens along with their premalignant counterparts were analysed for p53 alterations using immunohistochemistry, microdissection, polymerase chain reaction amplification, and DNA sequencing.Results-Malignant lesions from five of eight patients showed overexpression of p53 protein by immunohistochemistry. Upon DNA sequencing, two of these five specimens had p53 mutations. Of the five patients whose cancers showed p53 overexpression by immunohistochemistry, three had previous premalignant lesions that also had immunohistochemically detectable p53 protein. However, DNA sequencing showed that none of these three had mutations in the p53 gene. The remaining five premalignant lesions had no immunohistochemically detectable p53 protein.Conclusions-Some premalignant lesions have increased p53 protein which can be detected by staining with antibody to p53. This staining is not caused by mutations in p53 that are found in subsequent tumours at the same site. PMID- 16696021 TI - A rapid PCR ELISA for the detection of activated K-ras in colorectal cancer. AB - Aims-To develop a rapid PCR ELISA procedure for the detection of mutations in K ras in a microtitre plate format, and to evaluate the assay for the detection of these mutations in human colorectal cancer.Methods-An enriched PCR method was used with labelled primers, and PCR product was captured on GCN4 coated immunoassay plates. Detection of biotinylated mutant product was performed by colorimetric assay with streptavidin-horseradish peroxidase. The assay was used to determine K-ras status in a series of 60 human colorectal neoplasms, together with paired normal colonic mucosa. Results from gel electrophoretic analysis were compared with ELISA results.Results-The assay proved reliable in detecting K-ras mutations in DNA extracted from both fresh and paraffin embedded colorectal tumours. ELISA results were comparable with results from gel electrophoresis. Mutations of K-ras were detected in 16 of 48 adenocarcinomas and five of 12 adenomas but no mutations were detected in normal mucosa. There was a highly significant difference (p<0.0005) between optical density values for carcinomas with mutant K-ras and their paired normal data. Adenomas did not show the clear distinction between positive and negative results seen with carcinomas.Conclusions-This assay provides a rapid and reliable means of detecting mutations in codon 12 of the K-ras oncogene. The single tube format colorimetric analysis in microtitre plates and clear discrimination between mutant and wild type genes makes the assay suitable for automation. The occurrence of intermediate results in the case of adenomas provides support for the hypothesis that mutations of K-ras occur early in the course of colorectal carcinogenesis. PMID- 16696022 TI - Calcium binding and concomitant changes in the structure and heat stability of calprotectin (L1 protein). AB - Aim-To obtain further data on the structure and conformation of calprotectin, a prominent leucocyte protein found in many species.Methods-The binding of Ca(2+) to calprotectin was studied by means of equilibrium dialysis using (45)Ca as tracer. The thermal stability and denaturation kinetics of calprotectin were studied by means of differential scanning calorimetry. Con-comitant alterations in optical activity resulting from different conditions were measured. A computer program calculated the parameters to fit different models of protein structure. Ultraviolet spectroscopy gave absorbtion spectra. Sedimentation velocity studies and molecular weight determinations by the low speed (sedimentation) equilibrium technique were performed.Results-A maximum of six calcium ions were bound per calprotectin molecule at 0.7 mM calcium chloride. The apparent dissociation constants were calculated. Ca(2+) ions increased the denaturation temperature by 26 degrees K. The enthalpy of denaturation was also increased by Ca(2+). Addition of Ca(2+) to the buffers caused a gradual change in the near UV circular dichroism spectrum, while only minor changes were seen at wavelengths of 210-240 nm. A gradual increase in the sedimentation coefficient was observed on addition of calcium chloride. The extinction coefficient at 279nm was determined: E(279)= 2.53.10(4) M(-1) cm(-1).Conclusions-Calprotectin can bind six calcium ions. Upon binding, the protein shows distinct conformational changes and increased thermal stability. The former may be of importance for its function, while the biological significance of the latter is unknown. PMID- 16696023 TI - Diagnosis of mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes in a Chinese family by PCR/restriction enzyme analysis. AB - The clinical presentation and the biochemical and molecular genetic findings are described in a 13 year old Chinese boy with MELAS (mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes). The diagnosis was initially suspected because of the characteristic clinical features and the strong family history of convulsions. Using polymerase chain reaction-restriction enzyme analysis, the heteroplasmic nt3243 A-->G mutation in mtDNA of peripheral blood leucocytes and a muscle sample was demonstrated. The oligosymptomatic relatives were then screened by this method and the degree of heteroplasmy was analysed. This appears to be the first report of a MELAS family in Hong Kong with this described mutation. Molecular genetic techniques are advantageous in the diagnosis of MELAS. PMID- 16696024 TI - Identification of a thrombin receptor with factor Xa receptor and tissue factor in human pancreatic carcinoma cells. AB - Venous thromboembolism is a common feature of pancreatic cancer. The underlying mechanism is unclear, but is likely to involve thrombin generation on the cell surface. Human pancreatic carcinoma cell lines (n=8) have been studied immmunohistochemically for the expression of tissue factor, factor Xa receptor, and thrombin receptor. Each antigen had a distinct pattern of immunoreactivity in cell membrane and cytoplasm. Tissue factor was predominantly localised to the membrane, whereas thrombin and factor Xa receptor were largely cytoplasmic in distribution. The results support the hypothesis of a coagulation cascade that starts with tissue factor, leads to thrombin generation, and might confer a biological advantage on tumour cells. PMID- 16696025 TI - Our first year. PMID- 16696026 TI - The mammalian mitogen activated protein kinase network. PMID- 16696027 TI - The role of human leucocyte antigen genes in the development of malignant disease. PMID- 16696028 TI - Demonstration of CD44 gene expression in cells from fine needle aspirates of breast lesions by the polymerase chain reaction. AB - Aim-To demonstrate the feasibility of studying specific gene expression in fine needle aspirates from clinical lesions. The reverse transcription/polymerase chain reaction (RT-PCR) technique was used to demonstrate CD44 gene expression in cells from diagnostic fine needle aspirates taken from patients attending the outpatient clinic for breast diseases.Methods-Polyadenylated RNA was extracted from the cells remaining in the syringe barrel after fine needle aspirate cytological diagnosis of 41 patients with breast lesions. Analysis of CD44 gene expression was performed by RT-PCR using primers flanking the site for insertion of the variant exons. The resulting products were separated on 1.2% agarose gels, transferred to nylon membranes using Southern blotting and hybridised with specific probes for standard (constitutive) and variant exons of this gene.Results-On hybridisation with the CD44 standard exon probe, the expected amplified product of approximately 482 base pairs was visualised in 22 of 41 samples examined. Further hybridisation with the "variant" exon probes (exons 7 (v2), 8 (v3), 9b (v4), 12 (v7), and 15 (v10)) on 12 of these samples showed the presence of large molecular variants in all of these samples. However, the expression pattern detected with the probes for exons 7 (v2), 8 (v3) and 9b (v4) differed among the patients.Conclusions-Expression of the standard and variant regions of the CD44 gene in cells remaining in the syringe after fine needle aspiration was demonstrated using RT-PCR. The 5' variant exon probes seemed to show different patterns of expression among the patients. Further studies are currently being conducted to determine whether there is any correlation between expression of the various components of this gene and cytological diagnosis. Using this method, it would be possible to study the expression of other candidate marker genes in breast cancer using fine needle aspirates. PMID- 16696029 TI - ras and p53 in the prediction of survival in Dukes' stage B colorectal carcinoma. AB - Aims-To determine possible associations between p53 allelic deletion, c-Ki-ras mutational activation, immunohistochemical detection of p53 and ras proteins, various clinicopathological variables, and patient outcome in 168 Dukes' stage B colorectal carcinomas.Methods-Allelic deletion at the p53 tumour suppressor gene locus was detected using polymerase chain reaction (PCR) based loss of heterozygosity (LOH) assays. Overexpressed proteins were detected using the CM1 polyclonal antibody. A PCR based assay was used to detect the presence of activating mutations at codon 12 of c-Ki-ras. Immunostaining was carried out using a monoclonal antibody to p21(ras).Results-p53 LOH, CM1 immunostaining, c-Ki ras mutational activation, and p21(ras) immunostaining were not predictive of survival by logrank analysis. Multivariate analysis using Cox regression did not predict survival in this group of tumours.Conclusions-Aberrations in ras and p53 are unlikely to play an important role in the subdivision of patients with Dukes' stage B colorectal carcinoma into more accurate prognostic strata. It is possible that later genetic events are more important in conferring a specific phenotype on the resultant Dukes' stage B tumour. PMID- 16696030 TI - Detection of c-Ki-ras mutations in bile samples from patients with pancreatic and biliary cancers. AB - Aim-To determine whether c-Ki-ras mutations can be detected in bile from patients with biliary strictures caused by pancreatic cancer and other biliary tumours, with a view to developing bile c-Ki-ras mutations as a non-invasive diagnostic marker of pancreatic cancer.Methods-Bile was collected from 89 subjects (47 controls (including patients with bile duct stones or benign stricture), 20 patients with pancreatic cancer, 11 with cholangiocarcinoma, five with ampullary cancer, and six with metastatic biliary obstruction) referred for endoscopic retrograde cholangiopancreatography. DNA was extracted from bile and c-Ki-ras codon 12 mutations were detected using PCR and a restriction enzyme digestion method.Results-c-Ki-ras mutations were detected in 10 (50%) of 20 patients with pancreatic cancer, in one (9%) of 11 with cholangiocarcinoma, and in two (33%) of six patients with metastatic biliary obstruction (primary tumours: colon and prostate). C-Ki-ras mutations were not detected in the controls and patients with ampullary cancer.Conclusions-The sensitivity of this test is too low at 50% to recommend its use clinically, but with refinement has potential as a diagnostic marker for pancreatic cancer. PMID- 16696031 TI - Enumeration of semen leucocytes by fluorescence in situ hybridisation technique. AB - Aim-To determine whether the fluorescent in situ hybridisation technique (FISH) using a total human DNA genomic probe can be used to enumerate semen leucocytes.Methods-Semen samples from five donors were subjected to a mild KC1 solution. These samples were then biotin labelled under FISH conditions using a total human DNA genomic probe and the leucocyte counts were determined. To check the accuracy of the technique a monoclonal antibody against the common leucocyte antigen CD45 [KC56(T-200)] served as a control. An isotypic control for [KC56(T 200)], the immunoglobulin [MsIgG1], served as a secondary control.Results-Semen leucocytes stained by the FISH technique were easily detected because of their distinct bright yellow colour, while the sperm cells were red. The leucocyte count ranged from 0.5 to 4.9 x 10(6) per ml of semen. KC56(T-200) and its isotypic control MsIgG1, which served as control for the FISH technique, accurately identified 94% and 97% of the semen leucocytes of a control donor, respectively.Conclusions-The FISH technique using a total human DNA probe can accurately and effectively enumerate the overall leucocyte population in semen. PMID- 16696032 TI - MDM-2 protein expression in nasopharyngeal carcinomas. Comparative study with p53 protein expression. AB - Aims-To investigate the immunohistochemical expression of MDM-2 protein in comparison with that of p53 protein in nasopharyngeal carcinomas.Methods-Formalin fixed, paraffin wax embedded tissue from 59 cases of nasopharyngeal carcinoma was stained by immunohistochemistry for MDM-2 and p53 proteins.Results-The tumours were divided histologically into seven cases of keratinising nasopharyngeal carcinoma (KNPC), 14 cases of non-keratinising nasopharyngeal carcinoma (NKNPC), and 38 cases of undifferentiated nasopharyngeal carcinoma (UNPC). MDM-2 nuclear expression was observed in 0/7 KNPC, 1/14 NKNPC, and 11/38 UNPC. p53 nuclear expression was observed in 1/7 KNPC, 2/14 NKNPC, and 15/38 UNPC. Parallel MDM-2 and p53 expression was found in 12 cases (11 UNPC and one NKNPC). Discordant MDM 2-/p53 + expression was found in six cases (four UNPC, one NKNPC, and one KNPC), and absence of expression of both proteins in the remaining 41 cases.Conclusions Expression of MDM-2 and p53 proteins may be associated with the level of tumour cell differentiation in nasopharyngeal carcinoma. Simultaneous expression of MDM 2/p53 in a proportion of UNPC suggests that MDM-2 protein may be responsible for stabilisation of p53 protein in these cases, in view of the previous demonstration of the p53 gene in germ line configuration. This could be important in the pathogenesis of these cases, since MDM-2 may deregulate the p53 dependent growth suppressive pathway. Discordant MDM-2-/p53 + expression in a few cases of nasopharyngeal carcinoma may reflect stabilisation of p53 protein by other proteins, or p53 mutations unable to activate MDM-2. PMID- 16696033 TI - Induction of intercellular adhesion molecule 1 and class II histocompatibility antigens in colorectal tumour cells expressing activated ras oncogene. AB - Aims-To determine whether there is a correlation between activation of the ras oncogene and the induction of MHC class II antigens and intercellular adhesion molecule 1 (ICAM-1) by interferon-gamma (IFN-gamma).Methods-Expression of class II antigens, ICAM-1 and intracellular ras oncoprotein (p21) in established colorectal cell lines and short term cultures of primary colorectal tumour cells was determined by flow cytometry and mutation in the ras gene by sequencing of amplified segments of the gene.Results-The cell lines showed a variation in their modulation of MHC class II antigens and ICAM-1, ranging from no induction to a 98 fold increase in class II antigen expression in the HT29 cell line. Previous work indicated that most tumours could not be induced to express class II antigens. Four of the five least inducible lines either contained mutant ras or highly expressed the oncoprotein. The four highly inducible cell lines all contained non mutant ras. Of the 21 tumours studied in primary culture, 10 were inducible, one of which contained mutant ras. Of the remaining non-inducible tumours, four were mutant.Conclusions-Correlations between ras activation and failure to respond to IFN-gamma could not be shown to be significant. Therefore, ras activation, and concomitant subversion of intracellular signalling pathways, is probably not the major determinant in failure to activate class II antigens and ICAM-1. PMID- 16696035 TI - Circulating IgG autoanti-IgE antibodies in atopic patients block the binding of IgE to its low affinity receptor (CD23). AB - Aims-To investigate the ability of circulating IgG autoanti-IgE antibodies from atopic rhinitis patients to block the binding of IgE to its low affinity receptor (FcepsilonRII), also termed CD23.Methods-This involved the use of a well validated flow cytometric method to detect inhibition of FITC labelled IgE binding to human B cells expressing CD23 (RPMI 8866 cell line).Results-Taking inhibition values greater than 20% as being significant, 15 out of 20 IgG anti IgE containing sera inhibited the binding of IgE-FITC to the RPMI 8866 cells. The inhibitory effect was recoverable in the IgG fraction of serum, but was not related to the titre of either IgG1 anti-IgE or IgG4 anti-IgE, thus suggesting that it might be related to epitope specificity. No such inhibition was demonstrable with rheumatoid sera containing autoanti-IgG (that is, rheumatoid factor), but lacking autoanti-IgE.Conclusions-The capacity of anti-IgE to block the binding of IgE to CD23 has important implications, particularly in terms of upregulation of IgE synthesis and the consequent perpetuation of the inflammatory response. PMID- 16696034 TI - IGF-II dependent autocrine growth in cell lines derived from renal tumours of childhood. AB - Aims-To determine the role of insulin-like growth factors (IGF) in the proliferation of tumour cells, by studying the mitogenic response to IGFs of three cell lines of differing phenotype established from both malignant rhabdoid and Wilms tumour, representing a range of cell types (GOS 4, G401, and T3/73).Methods-Production of IGF-II and IGF-I was measured by radioimmunoassay, and the presence of IGF binding protein complexes was observed by gel exclusion chromatography. Following growth analyses in serum-free media to ascertain the dependence of the cell lines on exogenous IGFs, the generation of autocrine growth was measured by a density dependence assay of proliferation in culture. Receptors were measured by radioligand cross linking and autocrine growth through these receptors assayed by the use of blocking antibodies.Results-While GOS 4 and G401 were able to proliferate in serum-free medium over a period of 5 d, T3/73 showed an absolute dependence on IGFs added daily at 1-10 ng/ml. Plating at clonal density showed that cell growth was directly density dependent in serum free medium. The serum independent proliferation of G401 and GOS 4 was blocked by the addition of an antibody to the type 1 IGF receptor (alpha-IR3) suggesting that the effects of autocrine factors are mediated through type 1 IGF receptors. S1 nuclease protection analysis indicated that all three cell lines produced significant amounts of mRNA derived mainly from the P3 IGF-II promoter, but transcripts for IGF-I were undetectable. Radioimmunoassay of IGFs from conditioned media showed that all the lines made assayable IGF-II (8.6, 8.4, and 6.1 ng/ml/24 h/10(6) cells for GOS 4, G401, and T3/73 respectively). The presence of species consistent with both type 1 and type II IGF receptors was demonstrated using radioligand binding to cell membranes followed by cross linking.Conclusions Autocrine IGF-II may contribute to the serum independence of GOS 4 and G401 cells, whereas T3/73 may depend on exogenous IGF-II for proliferation. PMID- 16696036 TI - The role of a nested polymerase chain reaction in the diagnosis of Pneumocystis carinii pneumonia. AB - Aim-To compare the techniques and results of a nested PCR and an immunofluorescence assay (IFA) for the detection of Pneumocystis carinii infection; to consider the role of the nested PCR in the diagnosis of P carinii pneumonia (PCP).Methods-Serial dilutions of two known P carinii positive samples were tested by IFA and PCR to determine their relative sensitivities. Seventy eight respiratory samples (15 from 11 patients with HIV infection/acquired immunodeficiency syndrome (AIDS) and 63 from 42 patients with other forms of immunodeficiency) were tested using both assays, and the costs and technical requirements of each assay were assessed.Results-The PCR had a greater relative sensitivity over the IFA of 2 x 10(1) to 2 x 10(3) fold in a postmortem lung sample and 2 x 10(5) to 2 x 10(6) fold in a bronchoalveolar lavage sample from a patient with PCP. P carinii was detected in all 15 samples from the patients with HIV/AIDS by both IFA and PCR. Of the 63 samples from the patients with immunodeficiencies other than HIV/AIDS, the PCR was more sensitive than IFA.Conclusions-The nested PCR is a more sensitive assay than the IFA. It may be useful in the diagnosis of PCP in patients with immunodeficiencies other than HIV/AIDS. Similarly, PCR may be of benefit for this patient group as less invasive specimens are needed. PCR has an increasing role to play in the diagnosis of PCP in the routine laboratory. PMID- 16696037 TI - Diagnosis, epidemiology and pathogenesis of bacterial infections in the molecular era. PMID- 16696038 TI - Complete structural characterisation of the human aryl hydrocarbon receptor gene. AB - Aims-To clone and characterise the complete structural gene for the human aryl hydrocarbon receptor (AhR). This gene, located on chromosome 7, encodes a cytosolic receptor protein which, upon activation by various xenobiotic ligands, translocates to the nucleus, where it acts as a specific transcription factor.Methods-Primers, based on the AhR cDNA sequence, were used in conjunction with recently developed long range PCR techniques to amplify contiguous sections of the cognate gene. The amplicons produced were then cloned and characterised. A cDNA probe was also used to screen a human P1 library.Results-Using the cDNA primers, DNA fragments which mapped the entire coding region of the gene were amplified and cloned. All but one of these fragments were amplified directly from human genomic DNA. The remaining fragment was amplified using DNA prepared from a P1 clone as the PCR template. This P1 clone, obtained by screening a human P1 library, also contained the entire Ah locus. Characterisation of amplified and cloned DNA fragments provided sufficient information for the construction of a complete structural map of the gene. This also included 150 base pairs of nucleotide sequence data at all intronic termini.Conclusions-These data indicate that the human AhR gene is about 50 kilobases long and contains 11 exons. The overall intron/exon structure of the human gene is homologous to that of the previously characterised mouse gene; however, it is probably some 20 kilobases larger. These results demonstrate the need for further characterisation and provide the data to facilitate this. PMID- 16696039 TI - Adhesion of platelets to chemotactically responsive and non-responsive neutrophils. AB - Aims-To investigate the heterotypic adhesion of unactivated platelets to chemotactically responsive (migrated) and non-responsive (non-migrated) polymorphonuclear neutrophils (PMN).Methods-Platelets and PMN were isolated from autologous, normal blood. Migrated and non-migrated PMN were separated after N formylmethionyl-leucylphenylalanine (FMLP) stimulation. Platelets were labelled with a fluorescent monoclonal antibody directed against CD41 (GPIIb-IIIa). Platelets (3 x 10(8)/ml) and PMN (3 x 10(6)/ml) were incubated together. Heterotypic cell adhesion was measured in isolated PMN and PMN co-incubated with platelets by flow cytometric analysis of platelet marker fluorescence in PMN gated events. Platelet-PMN adhesion was also visualised by fluorescence microscopy.Results-In studies of isolated PMN, contaminating platelets were bound to 16-34% of unstimulated PMN, 7-22% of stimulated PMN, 2-4% of migrated PMN, and 17-24% of non-migrated PMN. When platelets were co-incubated with migrated or non migrated PMN, 15-78% of PMN bound one or two platelets.Conclusions-Unactivated platelets adhere to isolated PMN in vitro. Fewer unactivated platelets were adhered to migrated PMN than to non-migrated PMN in isolated PMN preparations. These results indicate that platelets adhering to PMN are removed during PMN migration. PMID- 16696040 TI - Identification of Candida species in formalin fixed, paraffin wax embedded oral mucosa by sequencing of ribosomal DNA. AB - Aim-To identify Candida species in formalin fixed, paraffin wax embedded tissue by sequencing candidal rDNA.Methods-Target rDNA sequences were amplified by polymerase chain reaction (PCR) from fresh isolates of Candida and from 18 preserved oral mucosal tissue samples (16 cases of chronic hyperplastic candidiasis and two fibroepithelial polyps), shown histologically to contain Candida. Identification of Candida species within tissue was based on a comparison of the rDNA sequences obtained with those from the fresh isolates of Candida and those present in the GenBank database.Results-The PCR products obtained from 12 of the 18 tissue specimens studied were characteristic of Candida albicans. In two of these cases a second, larger PCR product was obtained and these sequences were characteristic of Candida glabrata.Conclusions-Candidal DNA was amplified successfully from formalin fixed, paraffin wax embedded tissue. Sequencing of the PCR product enabled identification of the Candida species present. PMID- 16696041 TI - Sequence analysis of immunoglobulin variable region genes that encode autoantibodies expressed by lymphomas of mucosa associated lymphoid tissue. AB - Aim-To determine whether the immunoglobulin genes used by three gastric mucosa associated lymphoid tissue type lymphomas with known autoreactivity are mutated from germline as mutation from germline is an indicator of exposure to a mutational mechanism which characteristically acts on B cells as they undergo a follicle centre response.Methods-Cell lines established from two cases of MALT type lymphoma secrete autoantibodies recognising follicular dendritic cells (one case) and basement membrane (one case). The immunoglobulin heavy chain variable region genes (IgV(H)) and light chain variable region genes (IgV(L)) used by these cell lines, and the IgV(H) genes from a third case recognising human IgG were sequenced.Results-All three cases studied had mutated IgV(H) genes, while the IgV(L) genes were unmutated.Conclusion-The presence of mutations in IgV(H) genes is consistent with the origin of gastric MALT type lymphomas from B cells which have traversed the lymphoid follicle. PMID- 16696042 TI - Establishment and characterisation of two cell lines derived from a primary adenocarcinoma of the duodenum. AB - Aims-To establish two cell lines from a primary duodenal adenocarcinoma; to describe the morphological, growth, ploidy, and immunophenotypic characteristics of these cell lines.Methods-The cell lines, designated DAC/S and DAC/E, were characterised using both in vitro and in vivo cell culture techniques, light and electron microscopy, immunocytochemistry, and FACS analyses.Results-Both cell lines have an epithelial origin, are aneuploid and display characteristics of transformed cells. The cell lines differ from each other in morphology, doubling time and serum requirements. These cell lines are anchorage dependent and do not grow in nude mice.Conclusions-DAC/S and DAC/E cell lines are derived from neoplastic epithelium and could provide in vitro model systems for future investigations of the cell and molecular biology of duodenal neoplasia. PMID- 16696043 TI - Expression of type III collagen mRNA in renal biopsy specimens of patients with idiopathic membranous glomerulonephritis. AB - Aim-To investigate the distribution of type III collagen in membranous glomerulonephritis (MGN); to identify the cells responsible for the synthesis of alpha1 (III) mRNA.method-The distribution of type III collagen was studied by immunohistochemistry in 10 renal biopsy specimens, histologically diagnosed as MGN, and five control renal tissue samples obtained at surgery. Synthesis of alpha1 (III) mRNA was detected by non-radioactive in situ hybridisation.Results On immunohistochemistry, type III collagen was not observed in the control glomeruli, but was present focally in the glomeruli in samples from patients with MGN. No specific hybridisation signal for alpha1 (III) mRNA was found in the control glomeruli on non-radioactive in situ hybridisation. By contrast, positive signals for alpha1 (III) chain mRNA were detected in glomerular epithelial cells and mesangial cells in MGN tissue samples.Conclusion-These data suggest that additional synthesis of type III collagen by intraglomerular cells contributes to the changes in the glomerular basement membrane characteristic of MGN. PMID- 16696044 TI - Accumulation of p53 protein in inverted transitional cell papilloma of the urinary bladder. AB - Aims-To study the possible accumulation of p53 protein in inverted papilloma of the urinary bladder.Methods-Formalin fixed, paraffin wax embedded sections from 14 cases of inverted papilloma of the urinary bladder were studied retrospectively. Accumulation of p53 was detected by immunohistochemistry using a mouse monoclonal antibody directed against p53. p53 protein reactivity was scored as follows: 0 = 10%; 1 = 10% to <30%; 2 = 30% to <50%; and 3 = >50% of cells p53 positive.Results-The 14 sections were scored as follows: 3 in four cases; 2 in four cases; 1 in one case; and 0 in five cases. Overall, nine (64%) of the 14 cases were positive for p53 protein.Conclusions-The accumulation of p53 protein in inverted papilloma of the urinary bladder suggests that p53 may have has an important role in the neoplastic process of this tumour. However, the benign nature of inverted papillomas suggests that p53 protein accumulation is not related to tumour invasiveness and metastasis. p53 reactivity cannot be used as a marker of malignancy for urothelial neoplasia. Further studies are required to determine the role of p53 protein in the oncogenesis of urothelial neoplasms. PMID- 16696045 TI - Cyclin D1 amplification and expression in human breast carcinoma: correlation with histological prognostic markers and oestrogen receptor expression. AB - Aims-To study the amplification of the Cyclin D1 gene (CCND1) in human breast carcinoma; to relate this to Cyclin D1 protein expression; to relate these parameters to recognised pathological prognostic factors, including oestrogen receptor (ER) status.Methods-DNA extracted from frozen sections of breast tumours (n = 36) was used for Southern blotting. Probes for CCND1, c-myc and the immunoglobulin heavy chain locus (IgH) were hybridised to tumour DNA. Immunocytochemical expression of Cyclin D1 protein and ER was studied in paraffin wax sections from the same tumours.Results-Amplification of CCND1 was observed in 11% (four of 36) of tumours studied. Over expression of Cyclin D1 protein was observed in 73% (30/41) of tumours. There was no correlation between recognised histological prognostic markers and either gene amplification or expression. However, a weak association was seen between Cyclin D1 expression and ER status.Conclusions-A disparity exists between locus amplification and over expression of Cyclin D1, suggesting the existence of another mechanism for raised protein expression. No significant correlation was detected between either Cyclin D1 amplification or over expression and established prognostic markers. PMID- 16696046 TI - Insulin-like growth factors and IGF binding proteins in cyst fluid from patients with craniopharyngioma prior to intracavitary irradiation with Yttrium and thereafter. AB - Aim-To examine a series of cyst fluid samples from patients with craniopharyngioma at various stages of treatment in order to evaluate the use of insulin-like growth factors (IGFs) and IGF binding proteins as tumour markers or indicators of successful treatment, or both.Methods-Cyst fluid samples were obtained by stereotactic puncture prior to the intracavitary application of (90)Yttrium and at subsequent occasions. Analysis was performed by gel chromatography, radio-immunoassays, binding studies, and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) with subsequent western blotting.Results-IGF-I, -II and IGF binding protein-1 concentrations were measured in three craniopharyngioma cyst fluid samples. Immunoreactive IGF-I and IGF binding protein-1 concentrations in these three samples were between 6 and 29 ng/ml, and 17 and 48 ng/ml, respectively. In contrast, the IGF-II concentrations measured in 19 cyst fluid samples from seven patients with craniopharyngioma at various stages of treatment were much higher at 25-671 ng/ml. SDS-PAGE and subsequent western blotting using [(125)I]IGF-II as the ligand gave bands with estimated molecular weights of 330, 220, 135, 96, 46, 43, 34, 29, and 13.5 kDa in one adult, and identical bands at 220, 41.5, 37.5, 32, and 19 kDa in three cyst fluid samples from three children with craniopharyngioma.Conclusions-These results suggest that IGFs and IGF binding proteins are secreted by craniopharyngiomas and that they may alter the growth characteristics of these tumours. Furthermore, the distinct pattern of IGF binding protein sizes might be used as a tool for the differential diagnosis of tumours of the central nervous system. PMID- 16696047 TI - Combined non-isotopic in situ hybridisation and indirect immunohistochemical analysis of hormone production in the rat pituitary gland. AB - An understanding of the intracellular relation between hormonal expression (storage) and gene expression (production) is essential for elucidating the functional status of the individual cells in endocrine tissue such as the pituitary gland. To this end, mRNA expression was visualised by using a combined in situ hybridisation and immunohistochemistry method in routinely processed, formalin fixed, paraffin wax embedded rat pituitaries. mRNA was detected by non isotopic in situ hybridisation (alkaline phosphatase antialkaline phosphatase method, with nitroblue tetrazolium and 5-bromo-4-chloro-3-indolylphosphate as substrates). Sections were then stained by using the immunoperoxidase method to demonstrate pituitary hormone expression. The specificity of the combined staining method was confirmed by staining adjacent sections separately. The antigenicity of rat growth hormone and prolactin was adequately preserved following hybridisation. In conclusion, this method is specific, easy to use and permits the determination of the functional status of individual cells. PMID- 16696048 TI - Tumour heterogeneity: a problem in biopsy assessment of the proliferation index of oesophageal adenocarcinomas. AB - Tumour heterogeneity may pose a problem when biopsy specimens are taken to measure proliferation (for example, in assessing response to therapy). Two "biopsy specimens" were taken from the centre and two from the edge of the luminal surface of 20 resected oesophageal adenocarcinomas. The proliferation index for each "biopsy specimen" was measured by counting Ki67 labelled nuclei in histological sections. The proliferation index was not associated with tumour differentiation or stage. There was site specific heterogeneity with a significant difference in proliferation index between the central (mean (SD) 36.4 (9.7)) and edge "biopsy specimens" (39.3 (9.9)). There was, however, a wide range of differences between pairs of "biopsy specimens" from both sites. In conclusion, if a tumour is to be sampled for measurement of the proliferation index before and after treatment, then the sequential biopsy specimens (preferably duplicated on each occasion) should be taken consistently from a leading edge of the lesion. PMID- 16696049 TI - Molecular and biochemical aspects of interphase nucleolar organiser regions. PMID- 16696050 TI - Principles of ploidy analysis by static cytometry. AB - Aims-To examine the basic assumptions made during DNA ploidy analysis of histological sections with an aim to eliminate methodological errors that have lead to conflicting results with this technique.Methods-A rat liver imprint and histological sections together with sections and whole nuclei cytospins of human breast tumour biopsy specimens were stained with azure A Schiff's reagent and used to investigate the effects of computer imaging, histological section thickness, nuclear volume, and shape corrections.Results-The rat liver imprint demonstrated a linear relation between mean nuclear transmittance and nuclear area for each of the three (2C, 4C, 8C) hepatocyte clusters. This finding was used to produce similar, proportional integrated optical density (IOD) measurements from rat liver sections by selecting only nuclei that were sectioned through their centres, as assumed by the mathematics for volume correction. The limitations of computer imaging necessitated an edge (glare) correction for each nucleus so that nuclei of different sizes could be analysed. Shape correction was required to analyse nuclei of different morphologies. Normal human lymphocytes, squamous epithelium and fibroblasts were all measured with similar IODs. DNA ploidy values obtained from 7 mum human breast tumour sections (using lymphocytes as controls) correlated well with those obtained from the whole nuclei cytospins from the same tissue blocks.Conclusions-With an improved understanding of the theoretical and technical aspects of ploidy analysis of tissue sections, reproducible and consistent results are possible. These results can be integrated into routine histopathology investigations alongside immunohistochemistry and molecular diagnostic techniques. PMID- 16696051 TI - Detection of clonal immunoglobulin gene rearrangements in the peripheral blood progenitor cells of patients with multiple myeloma: the potential role of purging with CD34 positive selection. AB - Aims-To determine the extent of clonal cell contamination of peripheral blood progenitor cell (PBPC) collections in patients with multiple myeloma (MM) and to assess the purging efficacy of CD34 positive selection.Methods-PBPC collections from 29 patients with MM were analysed for the presence of clonal immunoglobulin heavy chain (IgH) gene rearrangements with a fluorescence based PCR technique. In addition, the PBPC from eight of the 29 patients were "purged" by selection of CD34 positive haematopoietic progenitors with an avidin-biotin immunoabsorption column (Ceprate). In each case the unmanipulated PBPC, CD34 positive and waste fractions were all assessed for the presence of clonal IgH rearrangements.Results Clonal IgH rearrangements (identical with those demonstrated in diagnostic bone marrow samples) were demonstrated in 10 (35%) of 29 cases and seemed to be confined to those with significant residual bone marrow disease. Clonal rearrangements were evident in the PBPC of two of the eight patients who underwent CD34 selection; in both instances a "clonal purge" was seen as it was not possible to demonstrate the clonal rearrangement in the CD34 positive fraction. In four of the six remaining cases the normal polyclonal fingerprint could not be demonstrated in the CD34 positive fraction, which is consistent with a significant reduction in contaminating B cells.Conclusions-Clonal cells contaminate PBPC collections in a significant proportion of patients with MM and may be eliminated by CD34 positive selection. PMID- 16696052 TI - Use of non-radioactive detection in SSCP, direct DNA sequencing and LOH analysis. AB - Aims-To develop a protocol that is applicable to single strand conformation polymorphism (SSCP), direct sequencing and loss of heterozygosity analysis of DNA.Methods-The protocol is based on the detection of biotinylated DNA by a Streptavidin-alkaline phosphatase conjugate. Biotinylation of DNA was achieved by using 5'-end biotinylated primers for PCR. After polyacrylamide gel electrophoresis, the DNA fragments were transferred to a nylon membrane by contact blotting. Depending on the alkaline phosphatase substrate, DNA was visualised either colorimetrically or by chemiluminescence.Results-The method was verified by the identification and characterisation of p53 mutations by SSCP analysis and direct DNA sequencing, as well as the assessment of DNA loss in human lung carcinomas by microsatellite polymorphism allelotyping.Conclusions-The protocol is simple, does not require specialised equipment and would be particularly useful for laboratories with experience in Streptavidin-biotin methodology. PMID- 16696054 TI - Nomenclature for proteins: is calprotectin a proper name for the elusive myelomonocytic protein? PMID- 16696053 TI - The genetics of inherited colon cancer. PMID- 16696055 TI - Improved serodiagnosis of Lyme disease. AB - Aims-To determine whether enzyme linked immunosorbent assay (ELISA) results for Borrelia burgdorferi require confirmation by immunoblotting and how immunoblotting may best be used in the diagnosis of Lyme disease.Methods-Over one year, all referrals for Lyme disease to a district general hospital with a large tick population in its catchment area were tested by ELISA. Positive, low positive and negative serum samples were subjected to immunoblotting and the reactive bands analysed.Results-In total, 633 samples were received; 38 were ELISA positive and 97 low positive. More serum samples were from rural (n = 356) than from urban (n = 277) areas but a higher percentage of serum samples from urban areas were ELISA positive. The ELISA results were confirmed by immunoblotting in 15/38 positive samples but in only four of 37 with a low positive titre. An IgM positive blot required a 41 kDa band plus >/=1 specific band; for IgG a 41 kDa band plus >/=2 specific bands were necessary. Five serum samples were IgM positive with a 41 kDa plus one or more other specific bands. For IgG blots, the best discrimination was seen with the 21, 31, 46, and 92 kDa bands. Nonspecific, weakly reacting bands at 55, 60 and 67 kDa were frequently seen. Infection was confirmed in four of six patients with arthritis, but in only one of 10 patients with erythema chronicum migrans.Conclusions-ELISA alone is insufficient for diagnosis. All positive and low positive or negative serum samples with a good clinical history should be examined by immunoblotting. A higher percentage of modified ELISA positive than low positive results were confirmed. There are significant differences between European and American immunoblotting patterns. Local results show similarity to American results, highlighting the need for a local Borrelia isolate. PMID- 16696056 TI - p53 gene alterations and protein accumulation in colorectal cancer. AB - Aim-To correlate immunohistochemical staining with single strand conformation polymorphism (SSCP) analysis of the p53 gene in colorectal cancer in order to understand how the findings provided by the two techniques complement each other in defining p53 functional status.Methods-Frozen tumour tissue from 94 patients with colorectal cancer was studied for p53 protein accumulation and gene mutations. Accumulation of p53 protein was detected by immunohistochemistry using PAb1801 and BP53-12-1 monoclonal antibodies. The findings were then compared with SSCP analysis of exons 5 to 8 of the p53 gene. All cases with a positive result by SSCP analysis were confirmed by sequencing.Results-Nuclear staining was observed in 51 (54.2%) cases. SSCP analysis of the DNA amplified by PCR revealed that the electrophoretic pattern had shifted in 30 cases; sequence analysis confirmed the occurrence of a mutation in 29 cases and of a polymorphism in one. In 27 cases both assays gave a positive result, and in 40 both were negative; therefore, concordance between PCR-SSCP and immunohistochemistry was seen in 72% of cases.Conclusion-The data indicate that positive immunostaining corresponds with the presence of a mutation in most, but not all, cases studied; other mechanisms could be responsible for stabilisation and accumulation of p53 protein in the nucleus. Nonsense mutations which do not confer stability on the protein will not be detected by immunohistochemistry and false negative results can also occur with SSCP analysis. PMID- 16696057 TI - Differential expression of novH and CTGF in human glioma cell lines. AB - Aims-(1) To investigate the expression in human derived glioblastoma cell lines of two structurally related genes, novH (nephroblastoma overexpressed gene) and CTGF (connective tissue growth factor), which encode putative insulin-like growth factor binding proteins of a novel type. (2) To investigate whether the same transcription factors regulate CTGF and novH expression.Methods-Expression of novH and CTGF was analysed in 24 glioblastoma derived cell lines by northern blotting. The CTGF promoter region was characterised by nucleotide sequencing, RNase protection experiments, by transient transfections, and CAT assays.Results CTGF and novH mRNA levels differed in the glioma cell lines studied. NovH and CTGF genes were not co-expressed in all cell lines. The CTGF promoter region was highly conserved compared with the corresponding region in the mouse (FISP12) and exhibited in vitro transcriptional activity.Conclusions-Although the coding regions of novH and CTGF are highly homologous, their promoter regions are substantially different, suggesting that these two genes may be regulated by different mechanisms. Considering that novH and CTGF are likely to be, respectively, negative and positive regulators of growth and that some glioma cell lines expressing novH are not tumorigenic, expression of these two genes might represent a key element in determining the stage of differentiation or the malignant potential, or both, of some tumour cell lines. PMID- 16696059 TI - We're on the Web! PMID- 16696058 TI - Relation between deletion of chromosome 1p36 and DNA ploidy in breast carcinoma: an interphase cytogenetic study. AB - Aims-To investigate whether deletion of the 1p36 region of chromosome 1 is independent of DNA ploidy in breast cancer cells.Methods-Preparations of nuclei from 64 fresh primary breast tumours were studied using dual target fluorescence in situ hybridisation (FISH) combining probes specific for the 1q12 (pUC 1.77) and 1p36 (1p-79) regions of chromosome 1. Signals were counted in 100-300 nuclei and the percentage of cells showing fewer p1-79 than pUC 1.77 signals was measured in each sample. DNA ploidy was investigated by cytofluorimetry in 55 tumour samples.Results-Chromosome 1 aberrations were detected in 56 samples. There were fewer p1-79 than pUC 1.77 signals in 53 samples. The 1p36 region was deleted in 11 samples in which a single p1-79 signal was detected; seven of these samples were diploid. Abnormalities were found in 17/24 diploid and 30/31 aneuploid tumours.Conclusions-Chromosome 1 aberrations, including deletion of the 1p36 region, were observed in diploid breast tumours. Deletion of the 1p36 region may be an early event in tumorigenesis. Given the frequency and importance of chromosome 1 aberrations in the biological behaviour of breast tumours, FISH, used in conjunction with cytofluorimetry, may be helpful for determining prognosis in patients with diploid tumours. PMID- 16696061 TI - Molecular biology and the Internet. PMID- 16696060 TI - Protein glycosylation in cancer biology: an overview. PMID- 16696062 TI - Differential expression of CD44 variants among meningioma subtypes. AB - Aims/background-CD44 is a widely distributed cell surface molecule which has numerous isoforms generated by alternative splicing. The diverse functions related to the CD44 variants (CD44v) have been reported in various physiological and pathological conditions. The pattern of expression of CD44v among meningioma subtypes was investigated to ascertain whether CD44 variants play a role in a variety of biological processes, such as epithelial differentiation and extracranial metastasis.Methods-Twenty three meningiomas were studied immunohistochemically using novel antibodies directed against CD44 isoforms. Six of the 23 samples were analysed by reverse transcription polymerase chain reaction (RT-PCR), followed by Southern blotting with CD44v specific probes.Results-In meningothelial, fibrous and anaplastic meningiomas, a standard form of CD44 was detected by RT-PCR and was homogeneously expressed in tumour cells when studied immunohistochemically. CD44v was not detected in these subtypes. In secretory meningiomas, however, CD44v isoforms were strongly expressed in the cell clusters that produce secretory granules and also accumulated in the granules. The population of tumour cells immunopositive for CD44v was similar to that which stained with antibodies directed against carcinoembryonic antigen, epithelial membrane antigen and ezrin. On RT-PCR with Southern blotting, only the secretory type showed high level expression of CD44v.Conclusions-CD44v in meningiomas is expressed in relation to tumour cell differentiation towards the epithelial type. PMID- 16696063 TI - Distribution of CD44 messenger RNA in archival paraffin wax embedded tumours and normal tissues viewed by in situ hybridisation. AB - Aims-We have previously demonstrated the abnormal localisation of expression of the CD44 gene in carcinoma cells in cryostat sections of fresh frozen tumour tissues, using radioactive in situ hybridisation (RISH). In order to facilitate further analysis of the expression of this gene in a wider range of neoplastic and non-neoplastic conditions, we have developed a technique which can visualise its low copy number transcripts in archival paraffin wax embedded specimens.Methods-(35)S labelled riboprobes complementary to transcripts from the standard (CD44s) and variant (CD44v) regions of the gene were used on paraffin wax embedded sections of tumours and corresponding normal tissues of the colon, breast and uterine cervix.Results-Elevated levels of signals for CD44s and CD44v transcripts were observed in carcinoma cells relative to their non-neoplastic counterparts in all tissues examined.Conclusion-This method permits easy access to material which can be selected for suitability, handled at room temperature without degradation and relied upon to show good histological detail. Comparison of the results with those on frozen tissues showed similar distributions of signals. Furthermore, the resolution and morphological detail was improved in paraffin wax sections. PMID- 16696064 TI - Genes mediating programmed cell death: an immunohistochemical study of bcl-2, c myc and p53 expression in colorectal neoplasia. AB - Aims-To describe the expression of three genes involved in the regulation of cell proliferation and programmed cell death (apoptosis) in normal, dysplastic and malignant large bowel epithelium, and to relate any alterations to important biological and clinical variables.Methods-Immunohistochemistry was used to assess bcl-2, c-myc and p53 gene expression in 70 colorectal carcinomas, 36 adenomas and three samples of normal mucosa.Results-Bcl-2 and c-myc protein were detected in all samples of normal mucosa and most adenomas. P53 was never found in normal mucosa and was expressed in only 5% of adenomas. Sixty nine of 70 carcinomas expressed c-myc protein; p53 was found in 46% and bcl-2 was present in 35%. Bcl-2 expression correlated with a higher degree of tumour differentiation whereas the opposite was true for c-myc. Strong staining for c-myc protein predicted survival in univariate analysis. No correlation was found between p53 and bcl-2 expression.Conclusions-While c-myc and bcl-2 proteins are overexpressed at an early stage of the large bowel adenoma-carcinoma sequence, alterations to the p53 protein level only occur as a late event in large, highly dysplastic adenomas and carcinomas. Bcl-2 may therefore protect the growing adenoma against excessive programmed cell death and mutated p53 may play a similar role in carcinomas. In vitro there is a reciprocal relation between p53 and bcl-2 expression. This could not be confirmed in vivo. Similarly, there was no relation between bcl-2 and c myc status, despite evidence that these proteins cooperate to cause neoplastic transformation. C-myc may be a prognostic indicator in large bowel cancer. There is no evidence in the present series that bcl-2 status will affect survival. PMID- 16696065 TI - Gastrointestinal hormone mRNA expression in human colonic adenocarcinomas, hepatic metastases and cell lines. AB - Aims-(1) To investigate the expression of the four main hormones of the digestive tract by performing reverse transcription polymerase chain reaction (RT-PCR) on a series of samples, comprising tumoral and healthy colonic tissues, hepatic metastases and colonic cell line samples; and (2) to study the patterns of labelling obtained with serological and morphological markers.Methods-After extraction and reverse transcription, gastrin, somatostatin, cholecystokinin (CCK) and transforming growth factor alpha (TGFalpha) mRNAs were detected by PCR and nested PCR using specific primers. The corresponding proteins were detected by immunohistochemistry.Results-The cell lines expressed all four mRNAs. Gastrin mRNA was present in most tumoral and metastatic samples, while the somatostatin transcript was detected in all samples and was frequently overexpressed in the normal colon. TGFalpha mRNA was expressed systematically in tumours of the right and transverse colon, but not in those located in the left colon; the expression of CCK mRNA was systematically absent in the left colon.Conclusions-The data presented here shed some light on the transcriptional events involved in the production of the various hormones present in the gastrointestinal tract, in both healthy and tumoral tissues. The various mRNAs expressed in cell lines are therefore not systematically expressed in the human pathology. PMID- 16696066 TI - Effect of Wnt-1 antisense RNA on the outgrowth of a mammary adenocarcinoma cell line expressing that oncogene. AB - Aims-To investigate the effect of Wnt-1 antisense RNA on the outgrowth of a mammary tumour cell line expressing that oncogene.Methods-A plasmid (pMT 70), containing Wnt-1 cDNA, was cut with appropriate enzymes and inserted into a eukaryotic expression vector (pMAMneo). A mammary tumour cell line (CAC-L153) was transfected with the expression vector and cells with the vector in sense and antisense orientation were selected.Results-Tumour cells with the expression vector in the antisense orientation had a notable reduction in expression of Wnt 1 protein and a considerable reduction in tumour outgrowth compared with controls.Conclusions-The results indicate that the Wnt-1 proto-oncogene may be a possible target for antisense therapy. PMID- 16696068 TI - Nested PCR-SSCP assay for the detection of p53 mutations in paraffin wax embedded bone tumours: improvement of sensitivity and fidelity. AB - DNA extraction and PCR amplification from paraffin wax embedded bone tumour specimens present several difficulties, firstly, because of the abundant matrix they contain and, secondly, because decalcification often causes degradation of DNA. In this report, comparative studies were carried out to determine the most efficient method for DNA extraction and PCR amplification from such specimens. The results indicated that nested PCR produced appropriate strong reaction products with minimal background contamination. A method for DNA extraction from paraffin wax embedded bone tissue and a nested PCR-SSCP technique have been developed for use in such diagnostic specimens. PMID- 16696067 TI - Expression of p53 in leukoplakia and squamous cell carcinoma of the oral mucosa: correlation with expression of Ki67. AB - Aim-To study p53 expression in relation to proliferative status in normal and nondysplastic, dysplastic and malignant lesions of the oral mucosa.Method-The standard avidin-biotin complex (ABC) immunohistochemical staining method was used to study the expression of p53 and Ki67 on frozen sections of oral leukoplakias and carcinomas.Results-Of the leukoplakia and carcinoma samples, 70% expressed p53 in over 5% of cells. In normal mucosa less than 5% of cells expressed p53. The proliferation index, as assessed by expression of Ki67, was highest in the malignant lesions (43%) and lowest in normal mucosa (11%). Statistical analysis revealed that expression of both p53 and Ki67 was correlated significantly with the histopathological stage of the tumour. However, expression of p53 was not correlated with that of Ki67. In leukoplakia lesions with proliferative features p53 immunostaining was less intense than in non-proliferative lesions; this difference was statistically significant.Conclusions-These results emphasise the potential of Ki67 and p53 as biomarkers of carcinogenesis in oral cancer and may also serve as intermediate points for cancer prevention programmes, such as the oral chemopreventive trials. Factors other than p53 may have a more important role in the deregulation of proliferation in pre-malignant oral lesions. PMID- 16696069 TI - A rapid and efficient method for DNA extraction from paraffin wax embedded tissue for PCR amplification. AB - DNA from archival, formaldehyde fixed, paraffin wax embedded human tissue, suitable for amplification by the polymerase chain reaction (PCR), was obtained using a microwave method based on the capture of DNA by magnetic beads. Fragments of the alpha-1-antitrypsin gene (AAT) and the apolipoprotein E gene (APOE) were amplified successfully from human liver and brain tissue, respectively. This procedure provides a more rapid, simple and efficient method for reproducibly obtaining DNA from preserved tissue that has been kept in storage for up to 30 years. PMID- 16696070 TI - Postmortem diagnosis of Factor V Leiden from paraffin wax embedded tissue. AB - Activated protein C resistance resulting from Factor V Leiden is an important inherited thrombophilia disorder which is found in 3.5% of people in the UK. The genetic defect can be detected using the PCR and the diagnosis can be made postmortem from paraffin wax embedded tissue. The presence of Factor V Leiden should be sought in all cases of unexplained sudden death resulting from venous thromboembolism. PMID- 16696071 TI - Immunohistochemical detection of Ki67 antigen in formalin fixed, paraffin wax embedded sections. PMID- 16696072 TI - Karyotypic and molecular abnormalities in chronic lymphocytic leukaemia. PMID- 16696074 TI - c-erbB3 protein expression in ovarian cancer. AB - Aims-To study the prevalence of overexpression of c-erbB3 growth factor receptor in ovarian carcinomas and to analyse its relation to histological subtype, stage and grade of the tumours.Methods-Ninety eight ovarian carcinomas were evaluated immunohistochemically using the RTJ1 monoclonal antibody raised against a synthetic peptide, the sequence of which was derived from the cytoplasmic domain of the c-erbB3 protein.Results-Of the tumours, 16% (16/98) overexpressed c-erbB3 protein relative to normal ovarian epithelium, whereas 22% (22/98) were completely negative. There was a statistically significant association between overexpression and well differentiated grade.Conclusions-These findings suggest that c-erbB3 protein overexpression occurs in a significant proportion of ovarian cancers and is correlated with differentiation. Overexpression may merit further investigation as a potential prognostic indicator and as a target for new treatment. PMID- 16696073 TI - The invasiveness of Entamoeba histolytica - a continuing enigma. PMID- 16696075 TI - Increased expression of CD44v6 in endocrine pancreatic tumours but not in midgut carcinoid tumours. AB - Aims/background-To analyse the different isoforms of CD44 in various types of endocrine pancreatic and gut carcinoid tumours and to investigate the relation between their expression and tumour dissemination. This study was prompted by the recent observation that inappropriate splicing of the CD44 gene was correlated with tumour progression and metastasis formation in a number of human cancers.Methods-Expression of CD44 isoforms was studied in 38 endocrine pancreatic tumours and gut neuroendocrine tumours using antibodies directed against products of exons v3, v4-v5, v6, v7-v8 as well as against the standard CD44 molecule (CD44H). CD44 gene expression was also analysed by reverse transcription PCR (RT-PCR) in nine endocrine and seven carcinoid tumours.Results All gastrinomas except one (nine of 10) and about half of the other endocrine pancreatic tumours (seven of 15) expressed CD44v6. Most (10/11) midgut carcinoid tumours were CD44v6 negative, with no detectable immunostaining. CD44v3, CD44v4 v5 and CD44v7-v8 were not expressed in any of these tumours. CD44 mRNA analysis illustrated a complex splice pattern and expression of large CD44 isoforms in CD44v6 positive endocrine tumours, whereas the standard form only was detected in midgut carcinoid tumours. No correlation between CD44 variant expression and tumour metastasis was observed.Conclusions-CD44 variants encoding exon v6 are preferentially expressed both in gastrinomas and in most pancreatic endocrine tumours. In contrast to other tumours, the expression of CD44v6 in pancreatic neuroendocrine tumours does not seem to be correlated with tumour dissemination. PMID- 16696076 TI - AgNOR quantity in needle biopsy specimens of prostatic adenocarcinomas: correlation with proliferation state, Gleason score, clinical stage, and DNA content. AB - Aims-To define the relation between the quantity of silver stained nucleolar organiser regions (AgNORs) and histological grade, clinical stage, DNA content, and MIB-1 immunostaining in needle biopsy specimens of prostatic adenocarcinomas.Methods-Histological grade was determined according to the Gleason system. AgNOR quantity, DNA content and MIB-1 immunostaining were evaluated by image cytometry on routine histological sections stained with silver, Feulgen reaction and MIB-1 antibody, respectively.Results-The mean AgNOR area increased with increasing Gleason score. A significant difference was found in the AgNOR values between low, intermediate and high grade tumours. Patients with clinically localised tumour (stages A and B) had lower AgNOR values than patients with advanced disease (stages C and D), but the difference in the mean AgNOR values between the two groups was not statistically significant. Non diploid tumours had a significantly higher mean (SD) AgNOR area than diploid tumours (3.68 (1.04) mum(2)v 2.73 (0.60) mum(2), respectively), while no significant difference was observed in the mean AgNOR values between aneuploid and tetraploid tumours (3.68 (1.04) mum(2)v 3.70 (1.05) mum(2)). When AgNOR and MIB-1-PI values were compared using linear regression analysis, a highly significant correlation was found.Conclusions-These data demonstrate that AgNOR quantity reflects the proliferative potential of prostatic adenocarcinomas, and is significantly related to histological grade and DNA content. The ease of application on routine sections, maintaining the morphological integrity of the tissue, the ability to evaluate selected histological areas of limited size and objective quantification by image cytometry make the AgNOR method particularly suitable for cell kinetic analysis in prostatic needle biopsy specimens. PMID- 16696077 TI - Counting of apoptotic cells: a methodological study in invasive breast cancer. AB - Aims-To arrive at a reproducible sampling technique for counting apoptotic cells in tissue sections of invasive breast cancer that can serve as a protocol for further clinical studies.Methods-In 4 mum thick haematoxylin and eosin stained tissue sections of 12 breast cancers, apoptotic cells, recognised by strict morphological criteria, were counted in consecutive fields of vision at x1000 magnification in a marked area in the most poorly differentiated region of tumour. These counts were regarded as the gold standard. Subsequently, in a systematic sampling experiment, the number of fields that had to be counted to derive an acceptable coefficient of variation (CV) was determined. To compare counts at different magnifications, all fields of vision were also counted at x630 and x400. The intra- and inter-observer reproducibility was tested by repeated measurements at these magnifications in 10 systematically selected fields of vision.Results-Apoptosis seemed to be a rare event, affecting, on average, about 1% of tumour cells. Noticeable clustering of apoptotic cells was observed. The systematic sampling experiment showed that at x1000 magnification, the CV was improved by counting up to 20 fields. When comparing x400 and x630 magnifications with the x1000 magnification, the correlation coefficients were 0.88 and 0.87, respectively. However, the lower magnifications yielded lower counts. With regard to reproducibility, the intra-observer correlation coefficient was 0.91 at x630 and 0.76 at x400. The inter-observer correlation coefficient was 0.77 at x630 and 0.68 at x400.Conclusions-Apoptotic cells can be counted readily in haematoxylin and eosin stained tissue sections. However, a systematic sampling protocol must be followed and cells should be counted at a relatively high magnification to obtain acceptable reproducibility. The suggested protocol will permit further correlative and prognostic studies and the monitoring of the effects of treatment. PMID- 16696078 TI - Accumulation of allelic losses on chromosome 10 in human gliomas at recurrence. AB - Aims-To elucidate the implications of allelic loss on chromosome 10 in the malignant progression of human gliomas.Methods-Eight microsatellite loci (D10S249, D10S191, D10S210, D10S219, D10S246, D10S222, D10S221, and D10S212) were analysed for chromosomal deletions in histologically benign and malignant, including recurrent, gliomas. Of the 16 original tumours studied (two astrocytomas, nine anaplastic astrocytomas and five glioblastomas), the histological diagnosis at recurrence was anaplastic astrocytoma in six cases and glioblastoma in 10. Genomic DNA was extracted from formalin fixed, paraffin wax embedded sections. Samples of original and recurrent tumours were paired and amplified using PCR. Samples of histologically normal brain served as controls.Results-Of the original tumours, all five glioblastomas, five (56%) of nine anaplastic astrocytomas and none of the astrocytomas demonstrated loss of heterozygosity (LOH) on chromosome 10. Additional LOH was detected in the five cases of anaplastic astrocytoma that progressed to glioblastoma at recurrence. Additional LOH was not detected in the two cases of astrocytoma that progressed to anaplastic astrocytoma at recurrence. With the exception of one case, additional LOH was observed in the recurrent glioblastomas.Conclusion-LOH was observed at the loci of two adjacent microsatellite markers, D10S222 and D10S221 (10q23-q25), suggesting that this region on chromosome 10 is closely related to progression from anaplastic astrocytoma to glioblastoma. PMID- 16696079 TI - An assessment of the comparative utility of functional and molecular level analyses in the investigation of patients with thrombophilia. AB - Aim-To determine the relation of the low anticoagulant response phenotype with the Factor V Q506 (Leiden) mutation in a cohort of patients with thrombophilia.Methods-Fifty four patients with either a personal or family history of deep vein thrombosis were investigated both for their anticoagulant response by the activated protein C resistance test (APCR) and their genetic status in respect of the Leiden mutation by means of a PCR-RFLP method.Results Low APCR ratios do not necessarily predict possession of the Leiden mutation. Conversely, normal ratios do not exclude the mutation. Of 14 individuals with low APCR ratios, the Leiden mutation was absent in five. Of the remainder, three were heterozygous and six homozygous. Of nine heterozygote individuals, only three had low APCR ratios. All patients homozygous for the defect had low APCR ratios.Conclusions-These results lend further weight to the hypothesis that the APC resistant phenotype results from more than one genetic defect and indicate the value of combined functional and molecular investigations in all patients with thrombophilia. PMID- 16696080 TI - Lymphocytes, cytokines and adhesion molecules in chronic graft versus host disease. AB - Aims-To determine which inflammatory and immune pathways are implicated in the development of chronic graft versus host disease (GvHD) and whether differences between these pathways are responsible for the different presentations of chronic GvHD.Methods-Biopsy specimens of diseased and normal skin were obtained from patients presenting with lichen planus-like and sclerodermatous type chronic GvHD. Expression of epidermal cytokines, adhesion molecules and lymphoid surface markers was analysed by means of immunohistochemistry. Apoptosis was detected using the in situ nick endlabelling method.Results-In both GvHD lesion types, CD8+ cells predominated in the epidermis, whereas CD4+ cells were the most prevalentin the dermis. Apoptotickeratinocytes were found in diseased skin only and Fas antibodies labelled a considerable number of keratinocytes. The epidermis in both types of lesions expressed interleukin (IL) 1alpha, tumour necrosis factor (TNF) alpha and intercellular adhesion molecule (ICAM)-1, but dermal vascular cell adhesion molecule (VCAM)-1 expression was restricted to specimens of lichen planus-like GvHD. IL1alpha and E-selectin were expressed in normal looking skin of 55% and 80%, respectively, of patients with lichen planus-like GvHD.Conclusion-The similarity between expression of epidermal cytokines and adhesion molecules (with the exception of VCAM-1) and lymphocyte phenotype in lichen planus-like and sclerodermatous GvHD strongly suggests that the latter occurs as a consequence of the healing process. VCAM-1 distinguishes between lichen planus-like and sclerodermatous lesions. IL1alpha and E-selectin are potential early markers of chronic GvHD. PMID- 16696081 TI - Lymphotropic herpes virus (EBV, HHV-6, HHV-8) DNA sequences in HIV negative Castleman's disease. AB - Aim-To evaluate the possible involvement of lymphotropic herpes viruses in Castleman's disease.Methods-Archival formalin fixed, paraffin wax embedded biopsy specimens from 16 HIV negative patients (11 with localised and five of multicentric disease) were studied. Epstein-Barr virus (EBV), human herpes virus 6 (HHV-6) and human herpes virus-8 (HHV-8) DNA was detected using PCR. PCR was also used to characterise the EBV genomes and the clonal status of the lesions.Results-EBV sequences were identified in nine (56%) cases. The main EBV genotype detected was type 1. Two (12%) cases were positive for both HHV-6 and EBV sequences. HHV-8 sequences were detected in one case of localised Castleman's disease, the sequence of which differed from that of the HHV-8 prototype. No clonal immunoglobulin gene rearrangements were found.Conclusions-EBV DNA was detected in a substantial proportion of cases, suggesting that it may have a role in the pathogenesis of Castleman's disease, unlike HHV-6 which was detected rarely. This is the first report of HHV-8 specific sequences in the localised from of the disease. PMID- 16696082 TI - Detection of human papillomavirus in vulval carcinoma using semi-nested PCR and restriction enzyme typing: a rapid and sensitive technique. AB - Aims-To develop a highly sensitive technique for the reliable detection and typing of human papillomavirus (HPV) DNA in clinical tissue.Methods-A two step, semi-nested PCR was used with primers spanning the L1 region of the HPV genome and capable of detecting HPV DNA of all known HPV types. The clinical samples were typed by digestion of the 412 base pair PCR product with Rsa I, generating unique fragments for each HPV type. Thirteen samples were screened by this method, including nine vulval carcinoma samples and four wart samples from the penis and vulva.Results-Experiments using DNA extracted from HPV DNA positive cell lines-that is, CaSki (HPV type 16) and HeLa (HPV type 18) established that the technique could detect as few as 50 HPV copies and that the predicted Rsa I fragments from HPV types 16 and 18 were generated. The predicted 412 base pair fragment was observed for all 13 clinical samples subjected to semi-nested PCR. Rsa I digestion of the product of the second round of PCR permitted the positive identification of the HPV type in most cases.Conclusions-This technique provides an effective and rapid means of detecting HPV DNA, in most cases providing the HPV type. High risk HPV types were always detected in the nine vulval carcinoma samples analysed. The amount of tissue available from the biopsy specimens was small, confirming the sensitivity of the method. PMID- 16696083 TI - Interphase ribosomal RNA cistron staining in thyroid epithelial cells in Grave's disease, Hashimoto's thyroiditis and benign and malignant tumours of the thyroid gland. AB - Aim-To evaluate the expression of ribosomal cistrons in human thyroid epithelial cells (TECs) of patients with Grave's disease, Hashimoto's thyroiditis and benign and malignant tumours of the thyroid gland.Methods-TEC nucleoli were investigated in fine needle biopsy specimens from 10 controls, 39 patients with Grave's disease, 15 with Hashimoto's thyroiditis, 56 with benign, and 15 with malignant tumours of the thyroid. A one step silver staining method was applied. In most cases serum concentrations of thyroxine and triiodothyronine as well as goitre size were determined. In every case 100 TECs were evaluated for the mean numbers of nucleoli and for the average number of argyrophilic nucleolar organiser regions (AgNORs) per nucleus.Results-NORs were activated in all patients, but not in controls. The numbers of AgNORs in patients with Grave's disease were closely correlated with thyroxine or triiodothyronine, or both, concentrations and with the size of the thyroid. In patients with Hashimoto's thyroiditis about 30% of TECs nucleoli did not contain AgNORs, whereas others were heavily impregnated with silver. Compared with controls and benign tumours, the nucleoli of carcinomatous TECs were larger and irregular in shape. The mean number of AgNORs per nucleus in malignant cells was higher than that in their benign counterparts.Conclusions-The mechanism by which NORs are activated in TECs varies depending on the type of lesion. The higher AgNOR score in TECs from malignant tumours can be used to distinguish them from their benign counterparts. PMID- 16696084 TI - Counting apoptosis-why and how? PMID- 16696085 TI - Role of free radicals and antioxidants in the pathogenesis of the inflammatory periodontal diseases. PMID- 16696086 TI - Tumour infiltrating lymphocytes: insights into tumour immunology and potential therapeutic implications. PMID- 16696087 TI - Correlation between apoptosis, proliferation and bcl-2 expression in malignant non-Hodgkin's lymphoma. AB - Aim-To investigate whether clinical features of non-Hodgkin's lymphomas, at the time of first biopsy, correlate with studies of cell proliferation and cell death as well as with the level of bcl-2 expression.Methods-Bcl-2 expression, determined by immunocytochemistry, was compared with cell proliferation, measured using in situ hybridisation for histone mRNA, and cell death by apoptosis, measured using in situ end labelling for DNA cleavage.Results-Histone mRNA staining gave a labelling index of 30% of cells for reactive germinal centres, 5.2-13.5% of cells for low grade non-Hodgkin's lymphoma and 12.1-50.5% of cells for high grade non-Hodgkin's lymphoma. In situ end labelling gave a labelling index of 5.0-10.0% of cells for reactive germinal centres, 1.0-3.7% of cells for low grade non-Hodgkin's lymphoma and 4.7-13.5% of cells for high grade non Hodgkin's lymphoma. There was a positive correlation between apoptotic index and proliferation index. More cases of low grade than high grade non-Hodgkin's lymphoma expressed bcl-2. There was no correlation between apoptotic index and bcl-2 expression for high grade non-Hodgkin's lymphoma.Conclusions-The molecular mechanisms controlling cell proliferation and death in non-Hodgkin's lymphoma are complex, probably involving a range of genes, including bcl-2. A better understanding of resistance to cell death is needed if the clinical goal of tailoring cancer treatment to individual tumours is to be achieved. PMID- 16696088 TI - Simple method for pretreatment of tissue sections for the detection of apoptosis by in situ end-labelling and in situ nick translation. AB - Aims-To overcome the problems associated with proteolytic pretreatment of tissue sections for the detection of apoptosis.Methods-Formalin fixed, paraffin wax embedded tissue sections of reactive lymph nodes and biopsy specimens of Burkitt lymphoma were pretreated by pressure cooking for the detection of apoptosis using the in situ end-labelling and in situ nick translation methods.Results-The results achieved with the in situ end-labelling and nick translations methods were compared with those obtained using a novel anti-apoptosis specific protein (ASP) antibody. The staining patterns generated using the three methods were similar and consistent, although the ASP antibody seemed to be more sensitive and detected higher numbers of apoptotic cells within sections.Conclusions-Pressure cooking is advocated as an alternative method to proteolytic enzyme digestion for pretreating paraffin wax sections. It is reliable, inexpensive, reduces the need to optimise pretreatment variables for different tissues, and permits double immunostaining of sections. PMID- 16696089 TI - p53 protein expression in non-Hodgkin's lymphoma. Comparative study with the wild type p53 induced proteins mdm2 and p21/waf1. AB - Aims-To investigate the pattern of expression of p53 protein and two wild type p53 induced proteins (mdm2 and p21/waf1) as an indirect way of assessing p53 gene status in non-Hodgkin's lymphoma.Methods-Formalin fixed, paraffin wax embedded tissue from 87 cases of nodal non-Hodgkin's lymphoma, comprising 52 high grade and 35 low grade tumours, was stained by immunohistochemistry for p53, mdm2 and p21/waf1 proteins.Results-p53, mdm2 and waf1/p21 proteins were expressed in 36/52, 21/52 and 31/52 high grade and 3/35, 21/35 and 3/35 low grade non Hodgkin's lymphomas, respectively. Parallel p53/mdm2 protein expression was found in 23 cases (21 high grade and two low grade). These 23 cases were also positive for p21/waf1 protein expression. Discordant p53 positive/mdm2 negative protein expression was found in 16 cases (15 high grade and one low grade). Eleven (10 high grade and one low grade) of these 16 cases were p21/waf1 positive and the remaining five high grade non-Hodgkin's lymphomas were p21/waf1 negative. Mdm2 and p21/waf1 proteins were not expressed in the absence of p53 protein expression.Conclusions-p53, mdm2 and waf1/p21 protein expression is more frequently associated with aggressive histotypes of non-Hodgkin's lymphoma. Parallel expression of p53, mdm2 and p21 proteins may represent non-Hodgkin's lymphomas with a wild type p53 gene as mdm2 and p21 proteins can be induced by the wild type gene. In these cases p53 protein expression may result from stabilisation via complex formation with the mdm2 protein. This could be important in the pathogenesis of these cases as mdm2 may deregulate the p53 dependent growth suppressive pathway. Discordant p53 positive/mdm2 negative/p21 negative protein expression may represent non-Hodgkin's lymphoma with p53 gene mutations unable to activate expression of mdm2 and p21 proteins. Non-Hodgkin's lymphoma with p53 positive/mdm2 negative/p21 positive protein expression may have either wild type p53 with deregulated mdm2 gene expression or mutated p53 gene with p53 independent p21 expression. PMID- 16696090 TI - Assessment of intra-tumoral karyotypic heterogeneity by interphase cytogenetics in paraffin wax sections. AB - Aim-To analyse the effect of sectioning on the assessment of karyotypic heterogeneity by interphase cytogenetics in paraffin wax embedded normal squamous epithelium and to apply the principles derived to invasive cervical carcinoma.Methods-Normal male (n = 5) and female (n = 5) squamous epithelia were hybridised with peri-centromeric repeat probes specific for chromosomes X (DXZ1) and 17 (D17Z1) individually and in combination to assess the effect of sectioning on mono-, di-, tri-, and tetrasomic populations. Section thickness, interobserver variation and variation between different areas of the epithelium were evaluated. Invasive squamous carcinomas of the cervix (n = 5) were then hybridised with the DXZ1 probe and intratumoral heterogeneity was assessed by comparison of signal distributions obtained from different areas.Results-The optimum section thickness for the assessment of normal epithelium was 6 mum. Variation in the expected signal number in the range 1-4 did not introduce artefactual heterogeneity at this section thickness. The sensitivity of this approach for the detection of minor subpopulations was calculated to be 13-16%, 17-18% and 10-11% for mono-, tri- and tetrasomic populations, respectively. Karyotypic heterogeneity was detected in two of the five tumours and, in one case where the populations where clustered morphologically, a minor population representing 18% was identified.Conclusions-Interphase cytogenetic analysis of sections from paraffin wax embedded material can be used for the detection of minor subpopulations in tumours. This approach will be of particular value in the assessment of the relation between human papillomavirus infection and tumour karyotype and in the analysis of intraepithelial neoplasia. PMID- 16696091 TI - Mutations in the Epstein-Barr virus latent membrane protein-1 (BNLF-1) gene in spontaneous lymphoblastoid cell lines: effect on in vitro transformation associated parameters and tumorigenicity in SCID and nude mice. AB - Aims-(1) To study the frequency of putative malignancy associated point mutations and a 30 base pair (bp) deletion in exon 3 of the C-terminus of the Epstein-Barr virus (EBV) encoded latent membrane protein (LMP)-1 (BNLF-1) gene in wild type EBV strains. (2) To assess the influence of these mutations on the tumorigenicity of lymphoblastoid cell lines (LCL).Methods-Eight spontaneous EBV (wild type) infected LCL were established from seven subjects. Deletions and single base mutations in the C-terminus of the BNLF-1 gene were demonstrated using bi directional solid phase dideoxy sequencing following PCR amplification of viral DNA from the LCL. Tumorigenicity of the LCL was assessed in SCID and nude mice. Serum dependent growth and ability to form colonies in soft agarose were assessed for representative LCL.Results-All LCL showed sequence differences compared with the prototypic EBV strain B95-8. The 30 bp deletion could be detected in three of eight LCL and a 69 bp deletion (including the 30 bp deletion) was identified in an additional LCL. A range of single base mutations (including those described previously in association with EBV related neoplasias) was also seen in some of the LCL. In transformation studies, the genetic variations did not seem to influence the in vitro behaviour of the LCL. In the tumorigenicity studies, the presence of the 30 bp deletion had no influence on the behaviour of the LCL which were, as expected, tumorigenic in SCID mice but not in nude mice. In contrast, the LCL carrying the 69 bp deletion was tumorigenic in both SCID and nude mice.Conclusions-Genetic changes described previously in the C-terminus of the LMP-1 gene in various malignancy derived EBV strains are also present frequently in wild type viruses and do not simply define tumour specific EBV strains. Changes within this region may, however, still be important for the tumorigenicity of LMP-1 and thus play a role in EBV oncogenesis. PMID- 16696092 TI - Activity of phosphoglycerate mutase and its isoenzymes in serum after acute myocardial infarction. AB - Aims/background-In humans there are three phosphoglycerate mutase (PGM, EC 5.4.12.1) isoenzymes (MM, MB and BB) which have similar distribution and developmental pathways to creatine kinase (CK, EC 2.7.3.2) isoenzymes. Total serum PGM activity increases in acute myocardial infarction with the same time course as creatine kinase activity. The present study was undertaken to determine changes in the activity of PGM and its isoenzymes after acute myocardial infarction.Methods-PGM activity was measured spectrophotometrically, by coupling the formation of 2-phosphoglycerate from 3-phosphoglycerate with enolase, pyruvate kinase and lactate dehydrogenase catalysed reactions. Inter- and intra assay reproducibility was assessed. PGM isoenzyme activities were measured using cellulose acetate electrophoresis.Results-Total PGM activity in serum was increased in patients with a confirmed diagnosis of acute myocardial infarction. PGM activity peaked 12 to 24 hours after the onset of symptoms and returned to normal values within 48 hours. Electrophoretic analysis of serum from healthy subjects showed a band corresponding to BB-PGM and two other artefactual bands that did not correspond to adenylate kinase. After myocardial infarction, BB-PGM activity increased and MB-PGM and MM-PGM could be detected. On immunoblot analysis, normal serum contained an inactive form of MM-PGM with a smaller molecular weight than that of PGM tissue isoenzymes.Conclusions-Total serum PGM activity increased in patients with acute myocardial infarction, following the same temporal course as creatine kinase activity. The increase in MM-PGM and MB PGM activities in these patients was not as high as expected. It is suggested that PGM isoenzymes, after release into the blood, undergo postsynthetic, probably proteolytic, transformation. PMID- 16696093 TI - Amphotericin B induced abnormalities in human platelets. AB - Aims-To investigate in vitro the effect of amphotericin B on platelets in order to understand poor platelet recovery in patients receiving platelet transfusions and amphotericin B simultaneously.Methods-Washed platelets were isolated from platelet concentrates and exposed to amphotericin B (4 mug/ml) for one hour. Platelet function was assessed by aggregation response to thrombin (0-0.6 U/ml), serotonin release, response to hypotonic stress, and mean platelet volume. The expression of surface membrane glycoprotein (GP) Ib-IX complex, GPIIb-IIIa complex and CD62P (P-selectin) was examined by flow cytometry using fluorescence labelled monoclonal antibodies. Heterotypic cell adhesion was measured in amphotericin B treated platelets coincubated with isolated, autologous polymorphonuclear leucocytes (PMN) by flow cytometric analysis.Results Amphotericin B induced platelet dysfunction. The rate of aggregation by thrombin, serotonin uptake and thrombin induced release of serotonin, and the response of platelets to hypotonic stress were inhibited. There was up to a two-fold increase in the mean platelet volume. The expression of platelet surface GPIb-IX and GPIIb IIIa was not affected. P-selectin, normally expressed only on the surface of activated platelets, was also expressed on unactivated platelets. Amphotericin B increased platelet adherence to PMN and the number of platelets bound per PMN.Conclusions-In vitro, amphotericin B induces P-selectin expression on the surface of unactivated platelets and increases platelet adhesion to PMN, which is exacerbated by storage. Platelet dysfunction resulting from exposure to amphotericin B may contribute to poor platelet recovery in vivo when amphotericin B is administered concomitantly with platelet transfusion. PMID- 16696094 TI - Rapid differential diagnosis of myxoid liposarcoma by fluorescence in situ hybridisation on cytological preparations. AB - In two cases of suspected myxoid liposarcoma, where chromosomal metaphase preparations were not available, fluorescence in situ hybridisation was performed on interphase nuclei of cytological preparations for the detection of the specific translocation, t(12;16), characteristic of this tumour and of trisomy 8, which is the most frequent secondary chromosome aberration. Probes directed against chromosomes 12 and 16 and against the centromeres of chromosomes 12 and 8 were hybridised on cell brushings and cytocentrifuge preparations. The finding of three painting domains of both chromosomes 12 and 16 and of only two signals with the centromeric probe directed against chromosome 12, suggested the presence of t(12;16) in both cases. In one case trisomy 8 was inferred from the occurrence of three centromere 8 signals. This approach can be used to detect specific chromosomal abnormalities when an urgent differential diagnosis is requested or when chromosome preparations are not available, or both. PMID- 16696095 TI - SSCP analysis of paraffin wax embedded tissues in a family with an atypical form of Fabry disease. AB - To investigate the distribution of a single base pair mutation within a family with one known case of Fabry disease, DNA from paraffin wax embedded necropsy material was studied using single-strand conformation polymorphism (SSCP) analysis. The proband, who presented with an atypical form of Fabry disease, had a G to A transition in exon 6 of the alpha-galactosidase A gene. This patient had mainly cardiac symptoms and late onset disease. Further cases of coronary disorders occurred in this family, including the proband's brother who died at 42 years of age of a cardiac disorder. Formalin fixed, paraffin wax embedded material from the brother and two more distant relatives was available for analysis. SSCP analysis showed that the proband's brother also carried the G to A transition. Thus, the atypical form of Fabry disease and unrelated cardiac diseases with similar clinical symptoms occurred within a single family. The variant form is rare but may account for a few of the numerous cases of cardiac disease in men and should be considered when clusters of cases of cardiac disease occur within a single family. PMID- 16696096 TI - Novel primer specific false terminations during DNA sequencing reactions: danger of inaccuracy of mutation analysis in molecular diagnostics. AB - The determination of nucleotide sequence is fundamental to the identification and molecular analysis of genes. Direct sequencing of PCR products is now becoming a commonplace procedure for haplotype analysis, and for defining mutations and polymorphism within genes, particularly for diagnostic purposes. A previously unrecognised phenomenon, primer related variability, observed in sequence data generated using Taq cycle sequencing and T7 Sequenase sequencing, is reported. This suggests that caution is necessary when interpreting DNA sequence data. This is particularly important in situations where treatment may be dependent on the accuracy of the molecular diagnosis. PMID- 16696097 TI - Applications of polymerase chain reaction-single stranded conformational polymorphism to microbiology. PMID- 16696098 TI - Cell adhesion molecules. PMID- 16696099 TI - Inhibitors of collagenase but not of gelatinase reduce cartilage explant proteoglycan breakdown despite only low levels of matrix metalloproteinase activity. AB - Aims-To investigate the level of matrix metalloproteinase activity during the time-course of cartilage explant proteoglycan breakdown; to determine the effects of selective small-molecule inhibitors of matrix metalloproteinases on proteoglycan degradation.Methods-The levels of matrix metalloproteinase activity in cartilage explant cultures and conditioned media were monitored by use of a quenched fluorescent substrate. The constants for inhibition of certain matrix metalloproteinases by a series of synthetic inhibitors were determined. Bovine and human cartilage explant cultures were treated with interleukin-1, tumor necrosis factor or retinoic acid and the amount of proteoglycan released into the culture medium in the absence and presence of the inhibitors was quantified. Control experiments, examining the inhibition of other proteinases, and investigating possible toxic or non-specific effects of the inhibitors, were carried out.Results-The profile of inhibition of proteoglycan release suggested the involvement of interstitial collagenase-like, rather than gelatinase- or possibly stromelysin-like, proteinases. No evidence was found for toxic or non specific mechanisms of inhibition. Very low levels of activity of the known matrix metalloproteinases were present during the time-course of aggrecan breakdown.Conclusions-A novel collagenase-like proteinase(s) may be involved in cartilage proteoglycan breakdown. Gelatinase-type matrix metalloproteinases do not seem to be involved in this process. Specific collagenase inhibitors may be therapeutically efficacious in the treatment of arthritis. PMID- 16696100 TI - Human papilloma virus detection by in situ hybridisation signal amplification based on biotinylated tyramine deposition. AB - Aim-To describe a method for amplifying human papilloma virus (HPV) in situ hybridisation (ISH) signals.Methods-Three human cervical cell lines, namely CaSKi, HeLa and SiHa, containing different copy numbers of integrated HPV DNA were studied. Following ISH, catalysed reporter deposition (CARD), based on the deposition of biotinylated tyramine at the location of the DNA probe, was used to amplify the ISH signal.Results-Using CARD-ISH, one to three HPV type 16 copies were detected in situ both in cell suspensions and paraffin wax sections of SiHa cells. CARD-ISH can also be used to detect oncogenic HPV DNA sequences, such as HPV types 16 and 18, in routinely processed formalin fixed, paraffin wax embedded cervical specimens.Conclusions-CARD-ISH is a fast and highly sensitive ISH method for the routine detection of low copy number HPV DNA sequences in cervical cell lines and routinely processed tissue sections. Application of this technology also enables the routine detection and cellular localisation of other viral DNA sequences present at copy numbers below the detection limit of conventional ISH methods. PMID- 16696101 TI - Direct in situ nucleic acid amplification: control of artefact and use of labelled primers. AB - Aims-To evaluate factors which ameliorate false positive artefacts with direct in situ PCR using labelled dNTPs; to investigate the use of labelled primers to overcome this artefact whilst maintaining sensitivity.Methods-Sections of measles (RNA virus) infected Vero cells with cytoplasmic signal or cytomegalovirus (DNA virus) infected fibroblasts with nuclear signal were collected. In situ PCR (or in situ RT-PCR) was carried out by methods permitting evaporation. Reagents or conditions which may control false positive artefacts using labelled dNTPs were investigated systematically. Labelled primers were tested to overcome artefacts, with adjuncts which improve sensitivity.Results-No reagent nor condition investigated was able to control the artefact with labelled dNTPs. Excessive digestion and incomplete DNAse treatments exacerbated the artefact, whereas novobiocin decreased both specific signal and artefact. However, the artefact was controlled by labelled primers, albeit with relatively low sensitivity. Sensitivity using labelled primers could be increased using alcohol fixation, albumin or Perfectmatch.Conclusions-A repair process is implicated for the artefact using labelled dNTPs. Excessive digestion or DNAse treatment may exacerbate DNA damage by disrupting histones or the DNA, respectively. Labelled primers control this artefact, albeit with reduced sensitivity, which may be improved by precipitation fixatives (alcohol) and reagents which enhance specific reaction. PMID- 16696102 TI - Significantly reduced expression of the proteoglycan decorin in Alzheimer's disease fibroblasts. AB - Aims-To investigate whether proteoglycan synthesis is altered in skin fibroblasts in patients with Alzheimer's disease compared with normal subjects.Methods-Cell lines obtained from donors with Alzheimer's disease and healthy controls were incubated with radioactive sulphate. The proteoglycans synthesised were determined and analysed by chromatographic, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and glycosaminoglycans-lyase treatment. The amount of decorin synthesised by each cell line was quantified using western blot analysis. Transcripts for human decorin were determined using northern blot analysis.Results-No significant changes in total sulphate incorporation and glycos-aminoglycan (GAG) composition were detected in the incubation media of these cells. However, chromatographic and SDS-PAGE analysis of the proteoglycans secreted by the cell lines showed that a dermatan sulphate proteoglycan of 150-125 kilodaltons was substantially reduced in Alzheimer's disease fibroblasts. The molecular characteristics of this proteoglycan correspond to decorin. Western blot analysis indicated that decorin was reduced in Alzheimer's disease incubation medium compared with normal medium. Northern blotting indicated that in Alzheimer's disease fibroblasts decorin transcripts were significantly reduced compared with normal fibroblasts. Glypican concentrations, a cell surface heparan sulphate proteoglycan, remained the same.Conclusions-These results strongly suggest that the expression and synthesis of decorin is affected in Alzheimer's disease skin fibroblasts. PMID- 16696103 TI - AgNOR clusters as a parameter of cell kinetics in chronic lymphocytic leukaemia. AB - Aims-To study correlations between the pattern of silver stained nucleolar organiser regions (AgNORs) in chronic lymphocytic leukaemia (CLL) and parameters of tumour kinetics. To investigate whether quantitation of the AgNOR pattern can be used to discriminate between patients with stable and progressive disease.Methods-Peripheral blood smears from 48 patients with CLL, classified as having either stable or progressive disease (Rai stage III or IV; bulky lymph nodes or massive splenomegaly; or peripheral lymphocytes >100 x 10(9)/1), were studied. For each patient, total tumour mass (TTM) and for patients undergoing a period of observation without treatment, the TTM duplication time (DT) and the lymphocyte doubling time (LDT) were calculated.Results-Four cell types could be distinguished according to their AgNOR pattern: (1) cells with a single cluster; (2) cells with a single compact nucleolus; (3) cells with two compact nucleoli; and (4) cells with several scattered dots. The percentage of cells with clusters was the AgNOR parameter which correlated best with TTM and LDT. Correlations were also seen between the proportion of cells with clusters and age and haemoglobin concentration. A significant correlation with DT could be detected only when age was kept constant. Linear discriminant analysis revealed that the percentage of cells with clusters was the most important prognostic factor. This alone classified 94% of the patients correctly (jackknive procedure) as either stable or progressive CLL.Conclusions-The percentage of circulating lymphocytes with clusters of AgNORs can be used as a parameter of tumour kinetics in CLL and helps to discriminate between patients with stable and progressive disease. For practical purposes, a value of more than 13% of cells with clusters is suggestive of progressive disease. PMID- 16696104 TI - Development of a rapid DNA screening procedure for the Factor V Leiden mutation. AB - Aim-To develop a rapid, simple and highly specific DNA screening procedure based on the amplification refractory mutation system (ARMS) to detect the Leiden mutation in whole blood.Methods-ARMS PCR amplification primers with additional mismatches at either -2 or -3, which greatly improves specificity, were constructed to detect the normal Factor V gene and the Leiden mutation in whole blood samples from patients with abnormal clotting results.Results-Construction of ARMS primers with either an additional mismatch at -2 or -3 at the 3' end of the primer could be used to detect the Leiden mutation in 0.5 mu1 whole blood in under three hours. Primers destabilised at position -3 could be used at a lower annealing temperature, which gave greater sensitivity and are now routinely used. A control set of primers was included in the same reaction to act as a positive control.Conclusions-This rapid and specific assay for the factor V Leiden mutation is a useful addition to the investigation of patients with or at risk from thrombovascular disease. PMID- 16696105 TI - Use of proteinase K for RT-PCR of cytokine mRNA in formalin fixed tissue. AB - Fresh tissue from cases of sudden infant death syndrome is becoming increasingly scarce and therefore researchers interesting in studying the aetiology of this syndrome have had to resort to archival tissue, usually in the form of paraffin wax sections. A simple method for isolating mRNA from formalin fixed, paraffin wax embedded material of sufficient purity for reverse transcription (RT)-PCR is described. Proteinase K treatment of formalin fixed, wax embedded tissue followed by RNA STAT-60 extraction was successful in isolating mRNA suitable for RT-PCR. Interleukin (IL)-1alpha, IL-6 and tumour necrosis factor (TNF) transcripts were amplified successfully from heart, but not thyroid, kidney or liver tissue, of a patient who died following rejection of a transplanted heart, and IL-1alpha, but not IL-6 or TNF, transcripts from lung tissue of a six month old baby who died of viral pneumonia. Transcripts of a housekeeping gene were detected in all tissues. This method should be useful for examining gene expression in archival material. PMID- 16696106 TI - Can I prevent travelers' diarrhea by taking antibiotics before my trip? PMID- 16696107 TI - Heart hurt by carbon monoxide poisoning. PMID- 16696108 TI - Coffee may slow the flow. PMID- 16696109 TI - Procedures for atrial fibrillation. PMID- 16696110 TI - Heavy metal and arsenic content in seabirds affected by the Prestige oil spill on the Galician coast (NW Spain). AB - Seabirds are top consumers in marine foodchains which offer opportunities to detect and assess the toxicological effects of different inorganic elements on the marine ecosystem. In order to provide baseline data concerning trace element levels in seabird species from NW Spain, zinc, copper, arsenic, chromium, lead, cadmium and mercury concentrations were analyzed in liver of three different seabird species (common guillemot, Atlantic puffin and razorbill) affected by the Prestige oil spill in September 2002 on the Galician coast. In general, with the exception of mercury, levels of all the analyzed elements were similar or lower in comparison with those reported for the same species in other Atlantic areas, and did not exceed levels indicative of increased environmental exposure. PMID- 16696111 TI - Abstracts of the 2006 ECNP (European College of Neuropsychopharmacology) Workshop on Neuropsychopharmacology for Young Scientists in Europe, Nice, France, 9-12 March 2006. PMID- 16696112 TI - Abstracts of the 90th Meeting of the German Society of Pathology, Berlin, Germany, 19-21 April 2006. PMID- 16696113 TI - Abstracts of the 8th Headache Congress of the European Headache Federation, Valencia, Spain, 26-29 April 2006. PMID- 16696115 TI - Abstracts of the 41st Annual Meeting of the European Association for the Study of the Liver, April 26-30, 2006, Vienna, Austria. PMID- 16696114 TI - Abstracts of the 19th International Conference on Antiviral Research (ICAR), San Juan, Puerto Rico, May 7-11, 2006. PMID- 16696116 TI - [Abstracts of the 87th German Radiology Congress, 24-27 May 2006, Berlin, Germany]. PMID- 16696117 TI - Reply to the letter to the editor on 'Scalp metastases and scalp cooling for chemotherapy-induced alopecia prevention', by C. Christodoulou, G. Tsakalos, E. Galani & D. V. Skarlos (doi:10.1093/annonc/mdj008). PMID- 16696118 TI - The state of advanced psychiatric mental health nursing practice. PMID- 16696119 TI - Respond to We Need to Talk feature in the October-December 2005 issue. PMID- 16696121 TI - New plan for tuberculosis. PMID- 16696120 TI - Intraoperative imaging of venous thrombosis in a kidney transplant by duplex sonography. PMID- 16696122 TI - What is your diagnosis? Post-traumatic segmental biliary cirrhosis. PMID- 16696123 TI - Image fusion involving ultrasound. PMID- 16696124 TI - Neurensin-1 expression in the mouse retina during postnatal development and in the cultured retinal neurons. AB - Neurensin-1/Neuro-p24 (previously named Neuro-p24) is a neuron-specific membrane protein that is localized particularly in neurites. Neurensin-1 is considered to play an essential role in neurite extension during nervous development, regeneration and plasticity. To understand what role Neurensin-1 plays in retinal differentiation, we examined Neurensin-1 distribution and gene expression pattern in the postnatally developing retina of the mouse, because the retina is an excellent model for nervous development. In the postnatal day (PD) 1 retina, intense Neurensin-1 immunoreactivity was found in the optic nerve fiber layer. Faint staining was seen in the ganglion cells, presumptive amacrine and horizontal cells. As the postnatal development proceeded, the optic fibers became more intensely stained in addition to other parts of the retina such as the ganglion cells, inner plexiform layer and horizontal cells. In PD 10 retinas, the horizontal cell processes showed a prominently stained configuration. As the retina developed further to attain maturity, the staining in the retina became less pronounced, although the optic nerves remained positively stained. The distribution of Neurensin-1 mRNA was consistent with these results and confirmed that the ganglion, amacrine and horizontal cells actively synthesize Neurensin-1 in the developing retina. In the retinal cell culture from newborn mice, two types of neural cells were stained for Neurensin-1, one of which showed long processes and appeared presumptive ganglion cells. These results suggest that Neurensin-1 plays a role in the fiber extension of the retinal neurons, as has been observed in other central nervous tissues, and indicate that the developing retina is a suitable experimental model for the analysis of Neurensin function, both in vivo and in vitro. PMID- 16696125 TI - Action of citicoline on rat retinal expression of extracellular-signal-regulated kinase (ERK1/2). AB - Citicoline is an essential endogenous intermediate in the biosynthesis of phosphatidylcholine, which acts as a therapeutic agent in models of central nervous system injury and neurodegenerative diseases. The present study investigated the effects of citicoline on extracellular-signal-regulated kinase 1/2 (ERK1/2) expression in the rat retina after kainic acid (KA) treatment. KA (6 nmol) was injected into the vitreous of the rat eyes. The animals were then injected intraperitoneally with citicoline (500 mg/kg) twice daily after the KA injection. The neuroprotective effects of citicoline were estimated by evaluating temporal changes in ERK1/2 using terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL), immunoblotting and immunohistochemical techniques. The expression of phosphorylated ERK1/2 was slightly decreased after 6 h, and significantly reduced after 12 h, in the rats receiving the KA injection plus citicoline treatment. Our results demonstrated that citicoline decreased the activation of ERK1/2 due to the KA treatment, suggesting that it exerts its neuroprotective activity by reducing the concentrations of proteins involved in apoptosis. PMID- 16696126 TI - Effects of herbimycin A in the pilocarpine model of temporal lobe epilepsy. AB - Pilocarpine-induced status epilepticus (SE) causes widespread tyrosine phosphorylation in the brain. It has been postulated that this intracellular signal may mediate potentially epileptogenic changes in the morphology and physiology of particular brain regions, including the hippocampus. The present study evaluated the effects of herbimycin A, a protein tyrosine kinase (PTK) inhibitor, over the acute (during which intense biochemical and electrophysiological activation occurs) and the chronic phase (characterized by spontaneous and recurrent epileptic seizures and the presence of synaptic reorganization, e.g., mossy fiber sprouting) of the pilocarpine model of epilepsy. The administration of a single dose of 1.74 nmol of herbimycin A (i.c.v., 5 microL) 5 min after the onset of SE did not change the acute behavioral manifestation of seizures despite significantly decreasing c-Fos immunoreactivity in different areas of the hippocampus and of the limbic cortex. Herbimycin-treated animals developed spontaneous recurrent seizures, as did control animals, with a similar latency for the appearance of the first seizure and similar seizure frequency. Neo-Timm staining revealed that all animals experiencing SE, regardless of whether or not injected with herbimycin, showed aberrant mossy fiber sprouting in the supragranular region of the dentate gyrus. Herbimycin did not obviously affect neuronal cell death as evaluated in Nissl stained sections. These results indicate that the PTK blockade achieved with the current dose of herbimycin reduced the acute c-Fos expression but failed to alter the spontaneous seizure frequency or to attenuate the morphological modifications triggered by the SE. PMID- 16696127 TI - Lip lesion. PMID- 16696129 TI - Epilepsy and psychiatric disorders. PMID- 16696128 TI - Drug treatment of bipolar disorder. PMID- 16696130 TI - Treating depressed children: limits of psychotherapy. PMID- 16696131 TI - As a man (or woman) grows older... PMID- 16696132 TI - Caregiver stress: when helpers need help. PMID- 16696133 TI - Women's Health Initiative: not over yet. The largest study of women's health has raised at least as many questions as it has answered. Stay tuned. PMID- 16696134 TI - Recognizing and treating basal cell carcinoma. PMID- 16696135 TI - Link found between abdominal fat and gallbladder surgery. PMID- 16696136 TI - Kegels hold up as urinary continence treatment. PMID- 16696137 TI - By the way, doctor. My mother and I have a friendly disagreement about fats for baking cookies. I say shortening is the unhealthiest because of its high trans fat content; my mother says butter or lard is worse. What's the verdict? PMID- 16696138 TI - [Diabetes in elderly patients: emergence of a disquieting problem]. PMID- 16696139 TI - [Psychological perceptions of body image]. PMID- 16696140 TI - Intensive insulin therapy in the medical ICU. PMID- 16696141 TI - Intensive insulin therapy in the medical ICU. PMID- 16696142 TI - Long-term outcome of renal transplantation from older donors. PMID- 16696143 TI - Long-term outcome of renal transplantation from older donors. PMID- 16696144 TI - Long-term outcome of renal transplantation from older donors. PMID- 16696145 TI - Coronary-artery stents. PMID- 16696146 TI - Coronary-artery stents. PMID- 16696147 TI - NXY-059 for acute ischemic stroke. PMID- 16696148 TI - Lenalidomide and venous thrombosis in multiple myeloma. PMID- 16696149 TI - Filing of applications and requirements for widow's and widower's benefits. Final rule. AB - We are revising one of our regulations to clarify that we will protect a title II claimant's filing date as of the date the claimant or other proper applicant on the claimant's behalf completes and transmits to the Social Security Administration (SSA) the Personal Information Identification data on the Internet Social Security Benefit Application (ISBA). This revision addresses an aspect of implementing the ISBA to provide certain rights to Internet filers that we afford to other filers. In addition, we are correcting one word in a different title II regulation. The revision is necessary to correctly reflect the circumstances under which a claimant for widow's or widower's benefits as the insured person's surviving divorced spouse would be considered "unmarried." PMID- 16696150 TI - Medicaid program; state allotments for payment of Medicare Part B premiums for qualifying individuals: federal fiscal year 2006. Interim final rule with comment period. AB - This interim final rule with comment period sets forth the methodology and process used to compute and issue each State's allotment for fiscal year (FY) 2006 and FY 2007 that is available to pay Medicare Part B premiums for qualifying individuals. It also provides the preliminary FY 2006 allotments determined under this methodology. PMID- 16696151 TI - The use of ingredients of potential public health concern. Compliance with the HACCP system regulations and request for comment. AB - The Food Safety and Inspection Service (FSIS) is publishing this document to inform establishments that prepare meat and poultry products of the need to ensure that they maintain proper control over the use of ingredients, especially those that present a potential public health concern, and over the ingredient labeling of their products. Establishments should ensure that their systems provide such control as part of their next reassessment of their HACCP systems. FSIS invites comments on the matters presented in this document. PMID- 16696152 TI - Medicare program; inpatient psychiatric facilities prospective payment system payment update for rate year beginning July 1, 2006 (RY 2007). Final rule. AB - This final rule updates the prospective payment rates for Medicare inpatient hospital services provided by inpatient psychiatric facilities (IPFs). These changes are applicable to IPF discharges occurring during the rate year beginning July 1, 2006 through June 30, 2007. In addition, we are adopting the new Office of Management and Budget (OMB) labor market area definitions for the purpose of geographic classification and the wage index. We are also making revisions to existing policies and implementing new polices. PMID- 16696153 TI - Blood vessels recovered with organs and intended for use in organ transplantation. Direct final rule. AB - The Health Resources and Services Administration (HRSA) and the Food and Drug Administration (FDA) are amending their regulations to consider as part of an organ those blood vessels recovered with the organ that are intended for use in organ transplantation (HRSA regulation); and to exclude such blood vessels from the definition of human cells, tissues, and cellular and tissue-based products (HCT/Ps) (FDA regulation). We (HRSA and FDA) are taking this action to provide that blood vessels recovered with organs and intended for use in organ transplantation are governed by the regulations pertaining to organs. The regulation of other recovered blood vessels remains unchanged. We believe that this change will eliminate the unnecessary burden resulting from an organ procurement organization's efforts to comply with both FDA and HRSA rules with respect to blood vessels (FDA jurisdiction) and organs (HRSA jurisdiction). We are issuing these amendments directly as a final rule because they are noncontroversial, and there is little likelihood that we will receive any significant adverse comments. Elsewhere in this issue of the Federal Register, we are publishing a companion proposed rule under our usual procedures for notice and comment in the event that we receive any significant adverse comments on the direct final rule. If we receive any significant adverse comments that warrant terminating the direct final rule, we will consider such comments on the proposed rule in developing the final rule. PMID- 16696154 TI - Medicare program; prospective payment system for long-term care hospitals RY 2007: annual payment rate updates, policy changes, and clarification. Final rule. AB - This final rule updates the annual payment rates for the Medicare prospective payment system (PPS) for inpatient hospital services provided by long-term care hospitals (LTCHs). The payment amounts and factors used to determine the updated Federal rates that are described in this final rule have been determined for the LTCH PPS rate year July 1, 2006 through June 30, 2007. The annual update of the long-term care diagnosis-related group (LTC-DRG) classifications and relative weights remains linked to the annual adjustments of the acute care hospital inpatient diagnosis-related group system, and will continue to be effective each October 1. The outlier threshold for July 1, 2006, through June 30, 2007, is also derived from the LTCH PPS rate year calculations. We are also finalizing policy changes and making clarifications. PMID- 16696155 TI - Outbreak news. Avian influenza, Egypt--update. PMID- 16696156 TI - Challenges in global immunization and the Global Immunization Vision and Strategy 2006-2015. PMID- 16696157 TI - Influenza. PMID- 16696158 TI - Teenagers with cancer: an improving picture. PMID- 16696159 TI - Postconsolidation immunotherapy in leukaemia remission. PMID- 16696160 TI - Regulation of RANKL might reduce bone metastases . PMID- 16696161 TI - COL-3 benefits patients with AIDS-related Kaposi's sarcoma. PMID- 16696162 TI - Dendritic-cell vaccination for metastatic melanoma? PMID- 16696163 TI - Potential prognostic markers in penile cancer. PMID- 16696164 TI - Improved survival and response to hepatic infusion. PMID- 16696165 TI - Monoclonal antibody aids colorectal-cancer treatment. PMID- 16696166 TI - Combination treatment improves breast-cancer survival. PMID- 16696167 TI - Alzheimer's drug helps brain function after radiotherapy. PMID- 16696169 TI - Genetic testing might miss some inherited mutations . PMID- 16696168 TI - Benefit of experimental vaccine for gastric cancer? PMID- 16696170 TI - Scientific meetings might affect prescribing. PMID- 16696171 TI - Robert I. Simon, MD: Of psychiatry, law, and books. PMID- 16696172 TI - Orthopaedic information: how to find it fast on the Internet. PMID- 16696173 TI - [Patient information. Lactose intolerance]. PMID- 16696174 TI - Current awareness in human psychopharmacology. PMID- 16696175 TI - Coumestrol, bisphenol-A, DDT, and TCDD modulation of interleukin-2 expression in activated CD+4 Jurkat T cells. AB - Endogenous estrogens are known to modulate several components of immune response, including interleukin-2 (IL-2) production. IL-2 is a cytokine that plays an important role in adaptive immune responses. These responses may be modulated by xenoestrogens such as coumestrol, bisphenol A (BPA), DDT, and TCDD. In this research, we examined the effects and potential mechanisms of action of these estrogenic compounds on IL-2 production in activated CD4+ Jurkat T cells. IL-2 production was analyzed by ELISA and Western Blot. At the transcriptional level, protein expression was examined by RT-PCR. Coumestrol, DDT and TCDD (but not BPA) significantly suppressed IL-2 production in activated CD4+ Jurkat T cells, at the transcriptional and translational levels. The transcriptional suppression of IL-2 was associated with decreased protein levels of NF-kappabeta, an important IL-2 positive transcription factor, without affecting the expression of Ikappa-Balpha protein expression, an important inhibitor of NF-kappabeta nuclear translocation. Although the direct mechanisms of xenoestrogens modulation of the immune system remain to be elucidated, coumestrol-, DDT- and TCDD-induced suppression of IL-2 may have ramifications for our understanding of the impact of xenoestrogens on health and disease. PMID- 16696176 TI - Cytotoxicity and transcriptional activation of stress genes in human liver carcinoma (HepG2) cells exposed to iprodione. AB - Iprodione (C13H13Cl2N3O3) is a broad spectrum dicarboximide fungicide used on a wide variety of crop diseases. It is used on vegetables, ornamentals, pome and stone fruit, root crops, cotton and sunflowers, to control a variety of fungal pests. Iprodione inhibits the germination of spores and the growth of the fungal mycelium. Experimental studies with mice have indicated that exposure to iprodione at dose levels 5 to 15 folds greater than the LOAEL for liver injury, induces microsomal enzyme activities, hepatocyte proliferation, hepatomegaly, centrilobular hypertrophy, diffuse hypertrophy, and an increase in lauric acid hydroxylation. Currently, there is no toxicological data available on human health effects associated with exposure to iprodione. In this research, we performed the MTT Assay for cell viability to assess the cytotoxicity of iprodione, and the CAT-Tox (L) assay to measure the induction of stress genes in thirteen recombinant cell lines generated from human liver carcinoma cells (HepG2). The cytotoxicity data indicated a strong concentration-response relationship with regard to iprodione toxicity. The percentages of cell viability were 100 +/- 0%, 128.0 +/- 41.4%, 97.5 +/- 37.7%, 70.1 +/- 35.4%, 33.5 +/- 16.1%, and 5.1 +/- 3.7% in 0, 31.3, 62.5, 125, 250, and 500 microg/mL, respectively. The LC50 was 208.3 +/- 83.3 microg/mL. Data obtained from the CAT-Tox (L) assay showed that iprodione is able to induce a significant number of stress genes in HepG2 cells. At 250 ug/mL exposure, the induction levels were 1.2 +/- 0.4, 50.1 +/- 17.8, 3.9 +/- 1.2, 16.8 +/- 7.2, 10.7 +/- 0.7, 1.8 +/- 0, 26.3 +/- 10.0, 7.2 +/- 2.4, 1.8 +/- 0.0, 6.8 +/- 1.3, 6.7 +/- 0.5, and 4.3 +/- 1.8 for CYP1A1, GSTYa, XRE, HMTIIA, c-fos, NF-kBRE, HSP70, CRE, RARE, GADD153, GADD45, and GRP78, respectively. These results indicate that the metabolism of iprodione involves Phase II biotransformation in the liver (XRE, GSTYa), and that this chemical has the potential to cause cell proliferation and/or inflammatory reactions (c-fos, NF-kB), proteotoxic effects (HSP70, GRP78), metabolic disruption (CRE), and DNA damage (GADD45, GADD153). PMID- 16696177 TI - Liver and renal function tests in artisans occupationally exposed to lead in mechanic village in Nnewi, Nigeria. AB - Additives in petroleum solvents have been reported to have adverse health implications. An evaluation study on some toxicological effects of occupational exposure to petroleum products (especially petrol which contains tetraethyl lead) amongst twenty five occupationally exposed artisans and twenty five graduate students of College of Health Sciences, Nnamdi Azikiwe University, Nnewi, Nigeria as controls, was carried out using the following biochemical markers: electrolytes, urea, uric acid, inorganic phosphorus, creatinine, zinc and blood lead, as well as the activities of alanine and aspartate aminotransferases, and alkaline phosphatase. The results showed that occupational exposure of human subjects to lead in petrol increases the concentrations of uric acid (357 +/- 123micro mol/L) and phosphate (1.5 +/- 0.5m mol/L) in exposed subjects compared with unexposed subjects (uric acid 228 +/- 105micro mol/L, phosphate 1.2 +/- 0.41m mol/L; p < 0.01 in both cases). Significantly lower activities were observed for alkaline phosphatase (66 +/- 18.9 iu/L). The activities of alanine aminotransferase (11.4 +/- 4.0 iu/L) and aspartate aminotransferase (15.8 +/- 4.4 iu/L) in occupationally exposed artisans were higher compared with unexposed subjects (alkaline phosphatase = 78 +/- 22.4 iu/L alanine aminotranferase = 6.8 +/- 2.7 iu/L, aspartate aminotranferase = 9.6 +/- 3.5i u+/-L; p < 0.01 in all cases). Occupational exposure of human subjects to lead significantly increased blood lead (59.6 +/- 15.9 microg/dL) and decreased plasma zinc (71.3 +/- 14.4 microg/L) in exposed compared with unexposed subjects (blood lead = 35 +/- 7 microg/dL, zinc = 108.4 +/- 16.9 microg/dL; p < 0.01). The results indicate that occupational exposure to lead in petrol may compromise liver and renal function. PMID- 16696178 TI - Changes in soil chemical properties and microbial activities in response to the fungicide Ridomil Gold plus copper. AB - The purpose of the study was to investigate changes of soil chemical and biological properties changes resulting from a single application of the fungicide Ridomil Gold plus copper (Ridomil Gold plus)(mefenoxam 6% + copper oxide 60%) at the following rates 0.25, 0.5, 1, 2, and 10 g m(-2). Selected chemical properties generally differed between fungicide rates over longer incubation periods. Microbial activity indices (available N, ammonification rates and specific enzymatic systems) were more sensitive indicators of change. Values of these indicators generally increased with incubation period and decreased or increased at high rates. Significant changes in P availability occurred after 90 days of incubation at rates > or = 1 g m(-2). Incorporation of the fungicide significantly increased NH4+ levels in soil after 75 days of incubation. These changes stimulated soil microbial activity as evidenced by increased ammonification rates especially at long-term exposure. Of the enzyme activities studied, dehydrogenase and beta-glucosidase activities were the most sensitive to ridomil gold plus. This sensitivity was more pronounced with the dehydrogenase activity. PMID- 16696179 TI - Toxicity of a mixture of 2,4-dichlorophenoxyacetic acid and monosoduim methanearsonate to the red swamp crawfish, Procambarus clarkii. AB - 2,4-dichlorophenoxyacetic acid and monosodium methanearsonate are often sold in commercial herbicide mixtures. Toxicity studies have been performed for each herbicide individually, but there is a dearth of information concerning the toxicity of these herbicides in a mixture. The following study examined the toxicity of a mixture of these two herbicides in the red swamp crawfish, Procambarus clarkii. 96-hour acute toxicity assays were performed to determine whether surfactant significantly altered the toxicity of these herbicides individually or in combination. Markings additive index was calculated to identify the interactions of the herbicide mixture. Surfactant was observed to significantly increase the toxicity of 2,4-dichlorophenoxyacetic acid and the toxicity of the herbicide mixture. The herbicide mixture alone displayed half the toxicity of the individual herbicides, but the mixture with surfactant was twice as toxic as the individual herbicides. The synergistic action of surfactant may be attributed to increased pesticide absorption across biological membranes. 2,4 dichlorophenoxyacetic acid and surfactant may also compromise gill function, increasing the sensitivity of the crawfish to herbicide toxicity. The antagonistic effects of the herbicide mixture in the absence of surfactant may be caused by competition of both herbicides for the same sites of activity. PMID- 16696180 TI - Molecular and cellular mechanisms associated with autoimmune diseases. AB - Evidence points to increases in the incidence and prevalence of several autoimmune diseases in the United States. As a result, the cost to public health from clinical management of autoimmune conditions is on the rise. The initiation and progression of autoimmune disturbances involves both genetic and environmental factors. Deficiencies in important proteins that normally participate in maintaining checks and balances within the internal milieu may render an individual prone to developing autoantibodies. Structural abnormalities or decline in normal levels of the pentraxins (serum amylase-P protein, the acute phase proteins, complement, and C-reactive proteins) have been shown to induce autoimmunity. Irregular transmission of information arising from multiple signal transduction pathways typically associated with the serine/threonine cascade routes of mitogen activating phosphorylation kinases, has also been found to induce autoimmunity. The kind of ligand/receptor interactions drives physical recruitment of different signals within the lymphocyte; these links define the quality and quantity of subsequent immune responses. CD95 or the Fas/Apo-1 and its ligand CD95L participate in regulating lymphocyte populations and therefore influence various aspects of immune responses. Mutational abnormalities resulting from synthesis of proteins by the CD95 and/or its ligand CD95L may result in alterations in the apoptotic pathways. Apoptosis may be completely inhibited, activated or partially stimulated. Modulation of apoptosis may lead to accumulation of self-antigens. Subsequently the immune system may be stimulated to react against self-molecules through lymphatic hyperplasia. This process may end up in proliferative disorders and enhanced susceptibility to autoimmune syndromes. This paper deals with mechanisms of autoimmunopathogenesis at the cellular and molecular levels. Emphasis is laid on the role of T and B cell receptor/ligand interactions, functions and malfunctions due to structural and quantitative alterations in T- B- cell clusterof antigen determinants. Genetically susceptible patients who develop spontaneous autoimmune diseases are examined and the etiological factors implicated in the initiation and subsequent dissemination of autoimmune diseases is discussed. PMID- 16696181 TI - Association of clinical complications with nutritional status and the prevalence of leukopenia among arsenic patients in Bangladesh. AB - This study conducted in Bangladesh reports the relationship of clinical complications with nutritional status and the prevalence of leukopenia among arsenic exposed patients living in the rural villages. A total of 115 exposed individuals diagnosed as arsenicosis patients were randomly selected from four known arsenic endemic villages, and age-matched 120 unexposed subjects were enrolled in the study program. The duration of arsenic exposure in about 37% of the patients was at least 10 yrs, while the population mean and range were 7.6 +/ 5.2 yrs, and 1 - 25 yrs, respectively. The mean arsenic concentrations in the drinking water for the exposed and unexposed (control) population were 218.1 microg/L and 11.3 microg/L, respectively. The spot urine sample of the arsenicosis patients contained an average of 234.6 microg/L arsenic. Although very few patients showed elevated WBC count, 16% had leukopenia (below normal count), and the whole population had significantly low WBC count than the control subjects. Prevalences of neutropenia and lymphocytosis were observed in patients with chronic exposure to high levels of arsenic in water. The body mass index was found to be lower than 18.5, the cut-off point for malnutrition (underweight), in about 28% of the arsenicosis cases compared to 15% of the controls. The monthly income and total calorie consumption per day showed the patients were underprivileged than the controls. Arsenical symptoms and complications were more severe in the nutritionally vulnerable (underweight) patients than the overweight ones. Also, the incidences of leukopenia and anaemia were more common in the female patients who were underweight. The findings of this research demonstrate that the poor nutritional status of patients increases the complications of chronic arsenic toxicity; suggest the possibility of other sources of arsenic contamination different from drinking water in the study area; and establish a higher prevalence of leukopenia and lymphocytosis in arsenicosis patients. PMID- 16696182 TI - Arsenic-induced genotoxic and cytotoxic effects in human keratinocytes, melanocytes and dendritic cells. AB - Arsenical keratosis and skin cancer are among the most common health effects associated with acute and chronic exposures to arsenic. This study examines the acute and chronic dose-responses of arsenic in established human cell lines using keratinocytes (HaCaT), melanocytes (CRL1675) and dendritic cells (THP-1 + A23187). Chronic conditions were established by treating the three cell lines with at least 8 passages in 0.2 microg/mL arsenic trioxide. Cytotoxicity was assessed using the fluorescein diacetate assay after 72 hrs of exposure. Single cell gel electrophoresis (Comet assay) was used to measure DNA damage. Acute exposure to arsenic had LD10 and LD25 values of 0.38 microg/mL and 3.0 microg/mL for keratinocytes; 0.19 microg/mL and 0.38 microg/mL for melanocytes; and 0.38 microg/mL and 0.75 microg/mL for dendritic cells. Cytotoxicity assays for chronically exposed cells resulted in LD10, and LD25 values of 0.4 microg/mL and 0.8 microg/mL for keratinocytes; 0.10 microg/mL and 0.20 microg/mL for melanocytes; and 0.10 microg/mL and 1.0 microg/mL for dendritic cells. The Comet assay showed that arsenic was highly genotoxic to the three cell lines. No significant differences (p > 0.05) in DNA cleavage were observed between acute and chronic exposures. In acute exposure arsenic genotoxicity was more severe with dendritic cells while melanocytes were more sensitive to arsenic cytotoxicity. Similarly, chronically exposed dendritic cells showed the maximum genotoxic damage while melanocytes were more sensitive to arsenic cytotoxicity. In conclusion, this research shows that arsenic is dermatotoxic, showing a high degree of genotoxicity and cytotoxicity to skin cells. PMID- 16696183 TI - Pentachlorophenol-induced cytotoxic, mitogenic, and endocrine-disrupting activities in channel catfish, Ictalurus punctatus. AB - Pentachlorophenol (PCP) is an organochlorine compound that has been widely used as a biocide in several industrial, agricultural, and domestic applications. Although it has been shown to induce systemic toxicity and carcinogenesis in several experimental studies, the literature is scarce regarding its toxic mechanisms of action at the cellular and molecular levels. Recent investigations in our laboratory have shown that PCP induces cytotoxicity and transcriptionally activates stress genes in human liver carcinoma (HepG2) cells [1]. In this research, we hypothesize that environmental exposure to PCP may trigger cytotoxic, mitogenic, and endocrine-disrupting activities in aquatic organisms including fish. To test this hypothesis, we carried out in vitro cultures of male channel catfish hepatocytes, and performed the fluorescein diacetate assay (FDA) to assess for cell viability, and the Western Blot analysis to assess for vitellogenin expression following exposure to PCP. Data obtained from FDA experiments indicated a strong dose-response relationship with respect to PCP cytotoxicity. Upon 48 hrs of exposure, the chemical dose required to cause 50% reduction in cell viability (LD50) was computed to be 1,987.0 +/- 9.6 microg PCP/mL. The NOAEL and LOAEL were 62.5 +/- 10.3 microg PCP/mL and 125.0+/-15.2 microg PCP/mL, respectively. At lower levels of exposure, PCP was found to be mitogenic, showing a strong dose- and time-dependent response with regard to cell proliferation. Western Blot analysis demonstrated the potential of PCP to cause endocrine-disrupting activity, as evidenced by the up regulation of the 125-kDa vitellogenin protein the hepatocytes of male channel catfish. PMID- 16696184 TI - Mitogenic and cytotoxic effects of pentachlorophenol to AML 12 mouse hepatocytes. AB - Pentachlorophenol (PCP), an organochlorine fungicide, is extensively used in the United States for the protection of wood products. Moreover, widespread agricultural, domestic, and industrial applications have caused PCP-contaminants to enter the food chain from the environment. There is accumulating evidence indicating that PCP is highly toxic to humans, and causes injury to major organs including the lung, liver, kidneys, heart, and brain. While PCP has been shown to induce systemic toxicity and carcinogenesis in several experimental studies, the literature is scarce regarding its toxic mechanisms of action. Recent investigations in our laboratory have shown that PCP exerts both cytotoxic and mitogenic effects in human liver carcinoma (HepG2) cells [1], and in primary culture of catfish hepatocytes [2]. In the present study, we hypothesized that PCP exposure will trigger similar cytotoxic and mitogenic responses in AML 12 Mouse hepatocytes. To test this hypothesis, we performed the MTT assay for cell viability in PCP-treated and control cells. Data obtained from this experiment indicated a biphasic response with respect to PCP toxicity; showing a hormosis effect characterized by mitogenicity at lower levels of exposure, and cytotoxicity at higher doses. Upon 48 hrs of exposure, PCP chemical doses required to cause 50% reduction in the viability (LC50) of AML 12 mouse hepatocytes was computed to be 16.0 + 2.0 microg/mL. These results indicate that, although the sensitivity to PCP toxicity varies from one cell line to another, its toxic mechanisms are similar across cell lines. PMID- 16696185 TI - Investigation into the nephrotoxicity of Nigerian bonny light crude oil in albino rats. AB - The effect of bonny-light crude oil was assessed in adult albino rats. The rats were administered with 200, 400, and 800 mg/kg body weight of the crude oil orally for 7 days. Fluid intake was measured daily, initial and final animal body was recorded. The toxic effects on the kidneys were assessed and histological studies carried out. The results revealed that the kidney cells were damaged; crude oil caused a destruction of the renal reserve capacity. There was a significant increase (p ? 0.05) in creatinine in the high dose group (800mg/kg), and a significant decrease (p ? 0.05) in urea concentration. Histological examination indicates that crude oil induced severe pathologic changes in the forms of necrosis and oedema. PMID- 16696186 TI - Freshwater microcosms-based assessment of eco-toxicological effects of a chemical effluent from the Pilcam industry in Cameroon. AB - We studied the acute toxicity of a raw effluent from a battery manufacturing plant (Pilcam) in Douala, Cameroon, to a freshwater fish (Oreochromis niloticus), and subsequently evaluated its sub-acute effects on water quality and the biota in freshwater microscosms. The acute toxicity test was based on 96 hrs static renewal bioassays that resulted in 96-h LC50 and LC90 values of 16 and 20.7% (v/v), respectively. The sub-acute experiments were conducted by exposing several species of aquatic organisms (plankton, macroinvertebrates and mollusks) to lower effluent concentrations [1.6%, 8.0%, 16% (v/v)] for six weeks, and monitoring their survival rates, as well as the physical and chemical characteristics of water. These concentrations were based on 10%, 50%, and 100% of the 96 h - median lethal concentrations (LC50) of the effluent to the freshwater fish, Oreochromis niloticus. Significant effects on functional parameters, such as, chlorophyll-a and total protein could not be demonstrated. However, the activity of alkaline phosphatase was significantly inhibited at all concentrations tested. Phytoplankton, zooplankton, macro-invertebrate communities and snails were negatively affected by the effluent application at concentrations ? 8% (v/v), with chlorophyta, ciliates, ostracoda, annelida, planaria and snails being the most sensitive groups. The snails were eliminated after 24 h exposure from microcosms treated with effluent at concentration ? 8% (v/v). Effluent exposure also caused significant effects on water quality parameters (DO, pH, hardness, conductivity, color, turbidity, ammonia) in general at concentrations ? 8% (v/v). Temperature and alkalinity were not significantly affected. Overall, data from this research indicate that a dilution of the Pilcam effluent down to 1.6% does not provide protection against chronic toxicity to aquatic organisms. Further studies are needed to determine the no observable adverse effect level (NOAEL), as well as a chronic reference concentration for this effluent. PMID- 16696187 TI - Effects of long-term exposure of the red swamp crawfish Procambarus clarkii to a mixture of two herbicides, 2,4-dichlorophenoxyacetic acid and monosodium methanearsonate, and associated human health risks. AB - 2,4-dichlorophenoxyacetic acid and monosodium methanearsonate are often sold in commercial mixtures. Bioconcentration studies have been performed for each of these herbicides individually, but little information exists concerning long-term exposure to a mixture of these herbicides. The following study examined the uptake of arsenic in crawfish after long-term exposure to this mixture, and the health risks associated with consumption of these crawfish. Bioconcentration and depuration experiments using a 50:50 by concentration mixture of the two herbicides, with and without surfactant, were performed to quantify how much arsenic is concentrated in the edible tissue of the crawfish. Of the three tissues (muscle, gill, and hepatopancreas) sampled hepatopancreas bioconcentrated the highest amount of arsenic. Surfactant significantly reduced this uptake but did not affect bioconcentration of arsenic into other tissues. Surfactant had no effect on depuration of arsenic from any of the tissues. Cooking lowered hepatopancreatic arsenic content, possibly as a result of structural changes in the hepatopancreas. Assessment of the human health risk associated with consuming these crawfish showed an exposure dose at the high end of consumption that was approximately twice the reference dose for arsenic. Cancer risks were averaged at approximately 7 extra tumors in a population of 10,000 and 6 extra tumors in a population of 10,000 resulting from a lifetime consumption of crawfish exposed to the herbicide mixture without and with surfactant, respectively. PMID- 16696188 TI - Metal ion effect on BOD exertion at different temperatures. AB - The toxic effect of metal ions like chromium (Cr3+), cobalt (Co2+), nickel (Ni2+), copper (Cu2+), cadmium (Cd2+) and lead (Pb2+) on biochemical oxygen demand (BOD) of synthetic wastewater samples has been studied at different temperatures i.e., 15 degrees C, 20 degrees C, 25 degrees C and 30 degrees C. Experiments were conducted for BOD exertion in presence (10 ppm of each metal ion) and in the absence of metal ions at different temperatures. Transition metal ions like Cr3+, Co2+, Ni2+ and Cu2+ show an increase in relative percentage inhibition with increasing atomic number. BOD inhibition in presence of Cd2+ and Pb2+ is relatively large. The metal ions under study are found to be highly toxic to microbes. PMID- 16696189 TI - Technical Report No. 38. Manufacturing chromatography systems postapproval changes (ChromPAC): chemistry, manufacturing, and controls documentation. PMID- 16696190 TI - Viability-based rapid microbiological methods for sterility testing and the need for identification of contamination. PMID- 16696191 TI - An alternative paclitaxel self-emulsifying microemulsion formulation: preparation, pharmacokinetic profile, and hypersensitivity evaluation. AB - Based on the clinical fact that paclitaxel injection (Taxol) frequently causes hypersensitivity reactions, an alternative paclitaxel self-emulsifying microemulsion was studied with phase diagrams, and the prescription of microemulsion formulation was optimized. Regarding Taxol, the pharmacokinetic parameters of microemulsion and hypersensitivity were investigated in rats and guinea pigs, respectively. The results showed that the self-emulsifying microemulsion was made up of tricaproin:tributyrin 1:1 as the oil phase, ethanol as assist, pluronic F68 and lecithin as emulsifier, and formed a mean diameter of 16 +/- 3 nm when diluted with saline. In the pharmacokinetic study, rats were administrated Taxol or paclitaxel microemulsion. Blood samples were collected at definite time intervals, and plasma concentrations of paclitaxel were determined by high-performance liquid chromatography. The area under the curve was significantly higher in the microemulsion group (33 mg.ml(-1).h) than that in the Taxol group (25 mg.ml(-1).h) (P < 0.01). The constant of transport rate of speed, K10 (0.55 h(-1)), was much smaller in the microemulsion group compared with the Taxol group (1.55 h(-l)). The mean retention time was 3.89 h in microemulsion group and 2.52 h in the Taxol group, showing the elimination rate was much slower in the former than in the latter. Compared with Taxol, the paclitaxel microemulsion caused less toxicity and had a longer circulation time in rats. PMID- 16696192 TI - Experimental study of the impact of annealing on ice structure and mass transfer parameters during freeze-drying of a pharmaceutical formulation. AB - The principal aim of this study is to evaluate the influence of annealing on structural properties and mass transfer parameters of a model formulation used for freeze-drying of pharmaceutical proteins. Characterization runs on ice morphology of the frozen material were performed in a cold chamber by direct optical microscopy using episcopic coaxial lighting. Consequently, it was possible to quantify the influence of annealing (temperature, duration) on ice morphology. Then, the pressure rise analysis method was used during primary drying to identify as a function of time both the dried layer mass transfer resistance values, R(p), and the sublimation front temperature, T(i), with or without annealing. Annealing turned out to accelerate sublimation rates by increasing ice crystal sizes of the frozen matrix, thus leading to lower values of R(p). Finally, secondary drying kinetics were studied by measuring the values of the desorption constant by a simple gravimetric method. In this study, annealing turned out to decrease desorption kinetics by a factor of two. PMID- 16696193 TI - Analysis of the purity of cetrimide by titrations. AB - The purity of cetrimide, trimethyl tetradecyl ammonium bromide (TTAB), which is an important preservative of many cosmetic and pharmaceutical products, was determined by three independent methods of titration. Traditionally, cetrimide was analysed by an assay method of the European Pharmacopoeia, which showed consistently a low purity of cetrimide with associated large standard deviations, however. A systematic 3% bias of the European Pharmacopoeia assay method was identified by comparing the result with results of two alternative methods of titration that exhibited high precision and high accuracy. Titration by perchloric acid showed a 99.69% +/- 0.05% purity of cetrimide, and titration by silver nitrate showed a 99.85% +/- 0.05% purity, while the traditional assay method predicted a purity of only 97.1% +/- 0.4%. It was found that the discrepancy could be identified as differences in selectivity during the extraction step of the European Pharmacopoeia assay method. The distribution coefficients between chloroform and water of cetrimide and the corresponding iodide species (TTAI) were thus determined as 2150 +/- 50 M(-1) and 68000 +/- 4000 M(-1), respectively. PMID- 16696194 TI - Microbial identification strategies in the pharmaceutical industry. AB - A commonly discussed question amongst pharmaceutical microbiologists, and the quality assurance organizations they serve, is what microbial isolates from the many microbial monitoring programs at a manufacturing facility need to be characterized, identified to genus, identified to species, or strain-typed? This article discusses the overall strategies that may be successfully applied to microbial identification in support of microbial monitoring of utilities, pharmaceutical ingredients, the manufacturing environment, and finished products. Emphasis in the article is given to the justification of the microbial identification program, selection of identification methods, and use of speciation in successful product failure investigations. PMID- 16696195 TI - The expanded application of most probable number to the quantitative evaluation of extremely low microbial count. AB - This paper is about the evaluation of the extremely low microbial counts from the field by expanding the most probable number (MPN) methodology when the data follow Poisson distribution in order to achieve more accurate estimation with limited number of data. Several data sets with extremely low microbial counts from pharmaceutical clean room and laboratory clean bench monitoring were found with good Poisson distribution fitness by chi2 test (P < 0.05), and the MPN calculations were conducted for these data by using the Halvorson and Ziegler equation. The MPN values are generally larger than the arithmetic average, indicating a higher sensitivity of the data assay. This approach is justified because Poisson distribution is the mathematical background of the MPN procedure, with the Halvorson and Ziegler equation as its foundation. It is considered that the MPN methodology has a potential application for quality control in the extremely low level microbial counting environment in the clean rooms levels ISO class 7 or above. Further studies on the precision of this method and development of a sampling plan based on careful mathematical analysis will help to refine the approach. PMID- 16696196 TI - [Why has the drug trial at Northwick Park Hospital turned out to a disaster?]. PMID- 16696197 TI - [Can cytostatics be effective in severe avian influenza? The disease picture has similarities with hemophagocytic lymphohistiocytosis]. PMID- 16696198 TI - [Distance pediatric radiotherapy. Telemedicine a good tool to be used for discussions, exchange of experiences and competence]. PMID- 16696200 TI - [E-health within the EU. From research to policy]. PMID- 16696199 TI - [Clinical research important in the future]. PMID- 16696202 TI - [Millions of newborn infants die unnecessarily. Simple means can result in big measures--and reduced perinatal mortality]. PMID- 16696201 TI - [Scientific misconduct, mistakes or something in between?]. PMID- 16696203 TI - [Drug committees should not recommend "off label" prescriptions]. PMID- 16696204 TI - [Peroral emergency treatment with injection solutions or tablets?]. PMID- 16696205 TI - [Gastrostomy a good aid--but no length of care changes using the different methods]. PMID- 16696206 TI - [Misleading information about cow milk hydrolysate]. PMID- 16696207 TI - [Exaggerated simplification of animal ethics]. PMID- 16696208 TI - [The discussion on WHO's definition of health is not new]. PMID- 16696209 TI - [Male circumcision protects against HIV]. PMID- 16696210 TI - [Tuberculosis dispensary in Uppsala--a 100-year anniversary]. PMID- 16696211 TI - [The new frontiers in the treatment of severe obesity]. PMID- 16696212 TI - [The "problem obesity": viewpoint of the internist]. AB - The prevalence of obesity has reached epidemic dimension in industrialized countries and it is known that obesity is associated with increased risk of cardiovascular morbidity and mortality. Commonly, obesity is defined by the Body Mass Index (BMI). However, BMI fails to consider body fat distribution. The relationship between the risk of metabolic-cardiovascular diseases and body fat distribution indices such as the waist-to-hip ratio (WHR) and the waist circumference, rather than measures of the degree of body fatness as expressed by BMI, has long been recognized. Recently, clinical and epidemiological research has found waist circumference to be the best anthropometric indicator of both total body fat and intra-abdominal fat mass. Android or visceral obesity is associated with metabolic syndrome and increased cardiovascular morbidity and mortality through a variety of molecular mechanisms possibly linking the metabolic syndrome to hemostatic and vascular abnormalities. Obesity guidelines suggest the need for weight reduction using behavioural change to reduce caloric intake and increasing physical activity. A realistic goal for weight reduction is to reduce body weight by 5% to 10% over a period of 6 to 12 months. Combined intervention of a low calories diet, increased physical activity, and behaviour therapy provides better outcomes for long-term weight reduction and weight maintenance than programs that use only one or two of these modalities. The drugs used to promote weight loss have been anorexic drugs or appetite suppressants. All classes of anorexic drugs affect neurotransmitters in the brain. The new agent sibutramine has norepinephrine and serotonin effects. Another new agent, orlistat, has a different mechanism of action, the reduction of fat absorption. Weight loss drugs approved by the FDA for long-term use may be useful as an adjunct to diet, physical activity and behaviour therapy for patients with a BMI of > or =30 with no concomitant obesity-related risk factors or diseases, and for patients with a BMI of > or =27 with concomitant obesity-related risk factors or diseases. PMID- 16696213 TI - [Obesity and immune function]. AB - The perspective of obesity as a low grade systemic inflammatory condition has triggered a new interest on the many overlapping areas between this pathology and the immune system. White adipose tissue production of proteins related to the immune function has shown that many of these adipokines are implied in the ethiopathogenesis of some of the major metabolic diseases such as diabetes, hypertension, cardiovascular diseases, which share with obesity an important role in the Metabolic Syndrome. Besides, dysregulation of immune system may be present due to a dysregulation in the factors produced by adipose tissue. Weight loss through diet or surgery has proved to be beneficial for the recovery of the physiological levels of some of these pro-inflammatory molecules, but other studies are needed to clarify to which extent its possible to pursue risk reduction by this way. PMID- 16696214 TI - [Diet therapy of severe obesity]. AB - In the last decades there has been a dramatic increase of obese subjects in many countries with the subsequent increase of health related problems, such as diabetes, hypertension, cardiovascular diseases. Even though several differential approaches, as media communication, guidelines formulation, have been performed in many different countries to solve this burden global health problem, no satisfactory results have been obtained. Personalized nutritional programs have been selected by obese subjects. Thus the correct approach would be to prescribe a caloric restricted diet in order to achieve long term weight loss in overweight or obese people. The aim in planning weight-reduction diets is that total food intake should meet recommended dietary allowance/adequate intake levels with the maintenance of the equilibrium among nutrients. If the nutritional approach fails or it is not sufficient to obtain significant weight loss, bariatric surgical intervention has been developed for the treatment of morbid-obese patients or severe obese patients with comorbidities as recommended by the American Society for Bariatric Surgery (A.S.B.S.) guidelines. PMID- 16696215 TI - [History and pathophysiologic analysis of the various techniques in bariatric surgery]. AB - Bariatric surgery for the treatment of morbid obesity or overweight refractory to medical therapy was born at the beginning of second half of the twentieth century, and its first steps were uncertain and with a not jet well definite purpose. In fact the main result to be pursued seemed to be simply the reduction of body weight, and any change of anatomy of the digestive tract able to reduce the absorbtion of nutrients was judged adequate. But very early the adverse consequences of malabsorption so obtained became evident, and other operations possibly free from those complications were devised and clinically tested. So aside the by-pass operations many other surgical procedures found their room, all of them aiming to fight the ever more diffuse obesity of the people. This historical review of the various surgical procedures attempted in these last sixty years for morbid obesity is very interesting for a better understanding of the problem and to have a solid basis for future rational choices. PMID- 16696216 TI - [Technical options in bariatric surgery and their mechanisms of action]. AB - In recent years, bariatric surgery has enjoyed a high level of approval. There are several technical and surgical options. Most of the departments of obesity surgery perform usually the same procedure, when just in a few center the procedure can be adapted depending on the patient need. In this paper the most popular technical choices in our country are presented based on the Consensus Conference that is taking place now in the Italian Society of Obesity Surgery (SICOB), in order to give our guidelines and policies. PMID- 16696217 TI - [Experience of many years with the Mason-Mc Lean procedure for the treatment of obesity]. AB - Bariatric surgery is still in search of a general agreement among surgeons for the more advisable technical procedure, and the various solutions actually overall performed are almost only on empirical basis. The main wanted result of such variety, of procedures is represented by the possibility to restore the body weight to physiological values without adverse effects. The Authors' experience is referred to 94 consecutive surgical procedures to treat obese patients and is completely homogenous, with all the patients treated by vertical gastroplasty, and the most of them are almost completely controlled till 24 months for clinical, biological, psychological and weight parameters. The complete absence of complications and the relevant compliance of all patients, without biological derangement from the normal values, correction of those preoperatively altered, and with the restoration of the body weight to the mean normal values of BMI, encourage the Authors to sustain the great advisability of the chosen technique. PMID- 16696218 TI - [Bariatric pacing. A new frontier for the treatment of severe obesity]. AB - A new bariatric procedure, gastric myo-electrical stimulation, has been developed. The Implantable Gastric Stimulator (IGS) induces satiety while avoiding the morbidity and mortality of the common restrictive malabsorptive or combination restrictive or malabsorptive procedure. The procedure does not alter normal anatomy. Advantages of IGS therapy are simplicity, a more rapid procedure, a safer procedure and the absence of nutritional side effects associated with some bariatric operations. The devices consists of a stimulation lead implanted in the gastric wall, connected to an electronic pulse generator (Transcend IGS, Transneuronix Inc, Mt. Arlington, NJ, SA) implanted subcutaneously into the abdominal wall. The lead is implanted by laparoscopy into the muscular layer of the lesser curvature of the stomach at the end of the pes anserinus area. The generator is connected (proper connection, was confirmed via radio frequency programming), and implanted subcutaneously. The mechanism of action of the IGS therapy requires a concomitant program of diet and lifestyle modifications in order to optimise the results. PMID- 16696219 TI - [Obesity and plastic surgery]. AB - The number of the bariatric surgical procedures to obtain a body weight loss is increasing, and will be higher in the years to come. The plastic surgeons are witnessing in parallel a meaningful growth of requests for surgical procedures to obtain a reshaping of body profiles after the procedures of bariatric surgery The surgical techniques of bariplastic surgery can be divided in three main groups: (1) Dermolipectomies (associated or not with lipoplastics). (2) Mammaplasties (with or without prosthetic implant). (3) Combined techniques (torsectomies). To correctly treat the morbid obese patients the multidisciplinary collaboration of a medical equip is of paramount importance. Bariplastic surgery after massive loss of weight has grown safe and efficacious if made on selected patients and in the hands of wide experienced surgeons. PMID- 16696220 TI - [Psychopathologic patterns in obesity]. AB - The problem of relationship between obesity and psychological distress is debated in the literature wherein a lot of studies exists with controversial results. The phenomenon of obesity is actually considered with criteria aimed to evaluate almost exclusively the weight excess. Even if such criteria have the advantage to permit a scientific communicability, in the clinical settings the focus of obesity involves medical as well psychiatric aspects. The psychological aspects that may have a relevant role in the development of obesity must be recognized and distinguished from those that may be a direct consequence of obesity itself. In fact, certain obese subjects (no-binge obese) may not experience any psychological distress during lifetime whereas other obese subjects (binge obese) may have a significant and highly distressing psychological suffering. Therefore, obese persons seem to represent a heterogeneous population with different adaptive characteristics who may show several and complex psychological mechanisms and distresses. A psychotherapeutic approach seems to be essential to treat such psychological distress that may heavily concur to the development and the maintenance of obesity. PMID- 16696222 TI - [Endoscopic palliation of obstructive jaundice caused by inoperable pancreatic cancer: personal experience]. AB - AIM OF THE STUDY: Few patients with pancreatic cancer are eligible for resection. In the remainder, estimation of prognosis is important to optimise various aspects of care, including palliation of biliary obstruction and trial of chemotherapy. The aim is to refer our personal experience about the palliation with endoscopic stenting in patients with unresectable pancreatic cancer. METHODS: The Authors reviewed retrospectively 132 patients affected by unresectable pancreatic cancer who underwent palliative interventions with endoscopic stenting from 2000 to 2004. RESULTS: Jaundice dramatically decreased in 86% of patients within 36 hours, in 12% within 48 hours and in 2% after two days. As complication were registered 3% of bleeding after endoscopic sphincterotomy, 2% mild acute post-ERCP pancreatitis and in 7% of patients transitory hyperamylasemia. The most frequent late complication was relapse of jaundice or cholangitis for stent clogging (51%) in a variable range fom 72.3 to 120.7 days, treated with stent substitution. No mortality ERCP-related was registered. CONCLUSION: Resection offers the only potentially curative approach to pancreatic cancer. The majority of patients are either too old, too ill with coexistent disease, or have a tumour that is undoubtedly inappropriate to resect. Thus for the vast majority an endoprosthesis to relieve the jaundice, is the preferred management, after a multidisciplinary approach. PMID- 16696221 TI - [Second tumours in patients with malignant neoplasms of the digestive apparatus. A retrospective study on 2406 cases]. AB - BACKGROUND: Incidence of a second tumor is elevated, between 2-10%, in patients previously affected by another tumor. Second tumor may concern the same organ, as a synchronous or metachronous way, or different organs. The present work is based on 4286 patients who underwent surgical treatment for gastric, small bowel, colorectal or pancreatic cancer at Surgical Unit 1- Minni- Bologna University and regards multiple primitive malignant tumor (T.M.M.P) of the II type (Moertel classification), such as different organs or tissues based second tumors; it also analyses overall incidence, sites, lesion number, latency period, and main association in our series. PATIENTS AND METHODS: 1742 patients with colorectal carcinoma, 1418 patients with gastric cancer, 91 patients with malignant tumour of the small bowel, 867 patients with pancreatic cancer and 68 with hepatocellular carcinoma have been included in the present study. A careful history, aimed at checking the presence of a previous tumor in a different site, was taken. One hundred eighty one patients (82 male and 99 female) reported tumor of a second type. Eleven cases of synchronous tumor and 170 cases of metachronous tumors were found, at a distance of 1-39 years. Moreover 7 patients reported more than one second tumor. Female genital tract tumors (37 cases), hematopoietic system tumors (34 cases), breast cancer (29 cases), and cutaneous tumor (20 cases) 9 were the most observed previous tumor. Incidence of a second tumor was more elevated in patients with small bowel neoplasms (18/91, 19.8%) than in patients with others tumors of gastrointestinal tract. CONCLUSIONS: Present study, although win part retrospective, point out an overall incidence of 4.3% of second tumors of the II type. Incidence of second tumours in patients with small bowel neoplasm, is particularly high; this confirm that small bowel plays a possible immunoprotective role and the immunodepression characterise patients with tumors in this site. PMID- 16696223 TI - A causal factors and treatment of obstructive ileus in 369 patients. AB - From 1998 to 2002, 369 patients (47.39% M, 52.61% F; mean age 67.9 yrs) were faced in the Authors' Department due to acute intestinal obstruction. The main reasons of obstruction were adhesions in 281 patients (76.15%), followed by malignant neoplasms of large intestine in 41 patients (11.11%) hernias in 9 patients (2.44%), Ischemic colitis (4.1%), bezoars (2.39%), bile stone (2.71%) and volvulus (1.1%). In this study the clinical and laboratorial investigation, the preoperative preparation and the applied surgical treatment of patients with obstructive ileus are discussed. Also it is stressed the importance of the proper preoperative support, as well as the convenient surgical confrontation of patients. PMID- 16696224 TI - [Complications in the treatment of cholangiolithiasis. Personal experience]. AB - Bile ducts lithiasis is a very common disease all over the world. In Italy prevalence is about 5 millions cases: 11% of the population. Choledocholithiasis concurrent with gallbladder stones frequency is estimated from 4 to 20%. Incidence is even higher in the elderly and in patients affected by chronic liver disease. The treatment of bile ducts lithiasis is still debated; several surgical strategies may be performed: (1) fully laparoscopic procedure; (2) endo laparoscopic sequential treatment; (3) sequential inverse treatment (endoscopy following video laparoscopic cholecystectomy); (4) combined endo-laparoscopic treatment simultaneously performed; (5) 'open" treatment. The authors refer their experience concerning a series of 172 patients who underwent endo-laparoscopic sequential treatment to amend bile duct and gallbladder lithiasis. Complications are pointed out, pending to demonstrate safeness and effectiveness of this strategy. PMID- 16696225 TI - [Anterior jugular vein cavernoma in a forthcoming rupture stage: a case report]. AB - The Authors present a very rare case of left anterior jugular vein cavernoma anterior jugular vein cavernoma in an over-seventy-year-old woman. The patient was visited already in a complicated status occurring as an increasing lateral neck mass. The case is so interesting for several reasons: for the peculiar lesion that belongs, in fact, to the uncommon group of the cavernous hemangiomas; for the exceptional localization on the anterior jugular vein; for the old age of the patient; last, but not least, for the unusual presentation of the forthcoming rupture. A speed healing without complications has been achieved thanks to the immediate diagnosis (it is important to suspect the lesion!), to the imaging techniques and to the surgical approach performed in emergency. PMID- 16696226 TI - Acute pancreatitis during liver hydatidosis: treatment with ERCP and endoscopic sphincterotomy. AB - INTRODUCTION: The Authors report on a case of a young woman who developed acute pancreatitis when affected by liver hydatidosis, successfully treated with endoscopic retrograde cholangiopancreatography (ERCP) and endoscopic sphincterotomy. METHODS: An endoscopic sphincterotomy was performed, with extraction of multiple hydatid membranes. OBSERVATIONS: Laboratory values returned to normal within 36 hours of the sphincterotomy. The patient was dismissed with oral therapy (Albendazole 400 mg bis in die for 4 months) and antibodies to Echinococcus were not detectable 1 month later. One year later, at ultrasound and CT the hydatid cyst was regressed and patient was still without symptoms. CONCLUSIONS: Hydatid membranes in the biliary tract should be considered as a potential cause of pancreatitis in patients with hydatidosis, even if it is a rare complication, caused by the obstruction of the distal part of common bile duct by fragments of hydatid membranes, scolices or daughter cysts. ERCP may be beneficial, but surgery remains the treatment of choice for treatment of liver hydatid cysts. PMID- 16696228 TI - [Two techniques for the transanal excision of rectal prolapse]. PMID- 16696227 TI - [Education in postgraduate surgical schools: the role of the surgical tutor as supervisor in the operating room]. AB - The Postgraduate Surgical education is in an era of transition, in order to create physicians with skills and attitudes needed by modern health care. Many studies have examined the impact of surgical tutoring in surgical residency programs in USA Medical Schools, while few experiences are reported from European Universities. The new Italian guidelines for post-graduate education require a structured clinical learning with the supervision of a tutor ("attending surgeon" for surgical residency); it is a challenge to describe the role of this teacher and educator, and to implement an effective evaluation of operating room teachers. Confidential survey was administered to 14 surgical residents of the Authors' University. Questions were related to their surgical activity and their perception of educational role of tutors in operating room and tutors' teaching behaviors. Residents pointed out five behaviors they perceive as signs of tutor excellence in clinical and operating room setting. According with studies from other Universities, residents need a tutor with competency but also with good teaching skills and a mature self-perception as educator. Faculty would provide training programs for surgeons in order to improve their teaching skills and behaviors. PMID- 16696229 TI - [White-collar criminals--a homogenous offender population? Reflections on typical and atypical "white-collar criminals"]. AB - Little has been written in German scientific literature on the personality structure of white-collar criminals. Often, the relevance of this level of investigation has downright been denied. Conventional psychopathology does not seem to be an appropriate approach to these character problems since there are not only deficits but also competences to be found which are useful while making a professional career. The author points out the inhomogeneity of this offender population and presents a case report of an atypical white-collar criminal. Over and above that, he introduces two psychological concepts which are apt to better describe the peculiarities of these individuals: Machiavellian intelligence is often the core competence when it comes to rising in hierarchies, whereas the newly defined psychopathy concept according to R. D. Hare makes plausible the moral and ethical failure of these offenders in their professional settings. PMID- 16696230 TI - [Domestic accident or child abuse?]. AB - While playing, a 33/4-year-old girl was hiding in a tumble dryer, which had been running before and started the drying process with rotation of the drum again after the girl had climbed into the machine and shut the door. The child suffered multiple haematomas, especially on the back and the lower arms, as well as second degree burns on body regions not covered by the clothing. The injury pattern was consistent with the properties of the appliance, and the initial suspicion that the child had been physically abused could not be maintained. PMID- 16696231 TI - [The significance of the legal term "corpse" in forensic medicine]. AB - The definition of the term "corpse" is relevant for medicine in general and most particularly so for forensic medicine, as it has direct consequences on practical work: legally, postmortems and autopsies may only be performed on corpses. With this in mind, the legal term "corpse" was examined on the basis of German State Law regulations and provisions. Approximately half the burial acts include a definition of "corpse", but even here there are certain variations. The definitions for "corpse" can be divided into four categories: 1) according to the lack of vital signs or sure signs of death 2) according to the degree of decomposition and/or cohesion of body parts 3) on the basis of severed body parts, and 4) terminological differentiation of stillbirths and neonatal fatalities from miscarriages. Laws vary greatly in their definition of the term "corpse". Therefore, the decision whether or not human remains or parts thereof qualify as a "corpse" should be left to medical discretion. PMID- 16696232 TI - [Gastric death as first described by Sehrt ("Sehrt'scher Magentod")--a review of the literature]. AB - The gastric death (Sehrt) is a rare form of death in the water with heterogeneous etiology. According to the first description, the pathogeneic mechanisms can be divided into two groups: asphyxiation after aspiration of vomited gastric contents and drowning due to hyperemia during digestion. In this paper, the authors discuss the different views on the causes of gastric death and evaluate relevant post-mortem findings. PMID- 16696233 TI - [Delayed pericardial tamponade following a stab wound to a branch of the right coronary artery]. AB - A 45-year-old man was attacked with a knife and suffered multiple stab wounds to the chest and abdomen. After surgical treatment and hospitalization for 9 days he was discharged in a stable condition. The following day he was found dead in his apartment. The autopsy revealed a pericardial tamponade caused by acute bleeding from an injured branch of the right coronary artery. This case shows that life threatening late complications in patients with penetrating cardiac wounds may occur despite an initially uneventful course. PMID- 16696234 TI - Treatment centre programme disarray as contracts axed. PMID- 16696235 TI - Job cuts are a 'necessary correction', says HR boss. PMID- 16696236 TI - NHS to miss key cancer target. PMID- 16696237 TI - Resubmitted plans face DoH scrutiny. PMID- 16696238 TI - Human resources. 'Consensus on the reform agenda has broken down'. PMID- 16696239 TI - On pay deals and incentives. PMID- 16696240 TI - HSJ debate. Sixty years on, how Bevan's vision is bearing up. PMID- 16696241 TI - Head for figures. Interview by Alex Nolan. PMID- 16696242 TI - Data collection. Designer data. PMID- 16696243 TI - Service improvement. In the know. PMID- 16696244 TI - Adult social care. Bridging the care divide. PMID- 16696245 TI - Finance. A fairer cut. PMID- 16696246 TI - Data briefing. Variations in length of hospital stays. PMID- 16696247 TI - Service redesign. Eight steps to better patient safety. PMID- 16696248 TI - On why PCTs need nurse leaders. PMID- 16696249 TI - Human resources. Top scorer. PMID- 16696250 TI - Staff well-being. Bully blues. PMID- 16696251 TI - Special section commentary: opportunities and challenges for human factors and ergonomics in enhancing patient safety. PMID- 16696252 TI - Describing nurses' work: combining quantitative and qualitative analysis. AB - OBJECTIVE: To better understand nursing activities and working conditions. BACKGROUND: Nursing practice involves astute clinical decision making and the competent delivery of nursing care procedures. To complete nursing procedures, nurses must simultaneously organize and reorganize priorities and manage changing clinical information for multiple patients. Few researchers have examined the specific nature of nurses' activities and the effect of the environment on their work. METHODS: Quantitative and qualitative methods were used to collect data on 7 nurses engaged in nursing activities. Examples of methods include link analysis, subject matter expert, task analysis, cognitive pathway, and "stacking." Data collection focused on how nurses managed priorities, changing clinical information, and interruptions. RESULTS: The "cognitive pathway" graphically depicts the nature of nurses' work. Specifically, it reveals the shifting of the nurse's attention from patient to patient and the occurrence of interruptions. "Stacking" shows the number of tasks a nurse must balance at any one time. On average, nurses had 10 or more activities waiting to be performed and experienced 3.4 interruptions/hr. CONCLUSION: These methods provide unique insight into the dynamic nature of patient care and nursing work. APPLICATION: Extension of the methods demonstrated here may be useful in guiding efforts to change the work of nursing to better provide quality care and less stressful work environments. PMID- 16696253 TI - Compliance with intended use of Bar Code Medication Administration in acute and long-term care: an observational study. AB - OBJECTIVE: To identify the types and extent of workaround strategies with the use of Bar Code Medication Administration (BCMA) in acute care and long-term care settings. BACKGROUND: Medication errors are the most commonly documented cause of adverse events in hospital settings. Scanning of bar codes to verify patient and medication information may reduce medication errors. METHOD: A prospective ethnographic study was conducted using targeted observation. Fifteen acute care and 13 long-term care nurses were directly observed during medication administration at small, medium, and large Veterans Administration hospitals to detect workaround strategies. RESULTS: Noncompliance with recommended practices was observed in all settings and facilities. A larger proportion of acute care nurses than long-term care nurses scanned bar-coded wristbands to identify patients (53% vs. 8%, p = .016). A larger proportion of acute care nurses than long-term care nurses administered bar-coded medications immediately after scanning (93% vs. 23%, p < .001). CONCLUSION: Workaround strategies were employed with BCMA that increased efficiency but created new potential paths to adverse events. There was a significant difference in the rate of use of workaround strategies between acute and long-term care. APPLICATION: The extent of workaround strategies varied by care setting and facility. BCMA should be tailored to the long-term care setting, including increasing the efficiency of use. Hospitals implementing bar coding should facilitate the intended use through equipment procurement, implementation, and quality improvement strategies. PMID- 16696254 TI - Toward the improvement of image-guided interventions for minimally invasive surgery: three factors that affect performance. AB - OBJECTIVES: The objectives were to measure the impact of specific features of imaging devices on tasks relevant to minimally invasive surgery (MIS) and to investigate cognitive and perceptual factors in such tasks. BACKGROUND: Although image-guided interventions used in MIS provide benefits for patients, they pose drawbacks for surgeons, including degraded depth perception and reduced field of view (FOV). It is important to identify design factors that affect performance. METHOD: In two navigation experiments, observers fed a borescope through an object until it reached a target. Task completion time and object shape judgments were measured. In a motion perception experiment, observers reported the direction of a line that moved behind an aperture. A motion illusion associated with reduced FOV was measured. RESULTS: Navigation through an object was faster when a preview of the object's exterior was provided. Judgments about the object's shape were more accurate with a preview (compared with none) and with active viewing (compared with passive viewing). The motion illusion decreased with a rectangular or rotating octagonal viewing aperture (compared with circular). CONCLUSIONS: Navigation performance may be enhanced when surgeons develop a mental model of the surgical environment, when surgeons (rather than assistants) control the camera, and when the shape of the image is designed to reduce visual illusions. APPLICATION: Unintentional contact between surgical tools and healthy tissues may be reduced during MIS when (a) visual aids permit surgeons to maintain a mental model of the surgical environment, (b) images are bound by noncircular apertures, and (c) surgeons manually control the camera. PMID- 16696255 TI - Labeling of medicines and patient safety: evaluating methods of reducing drug name confusion. AB - OBJECTIVE: We report three experiments evaluating the proposal that highlighting sections of drug names using uppercase ("tall man") lettering and/or color may reduce the confusability of similar drug names. BACKGROUND: Medication errors commonly involve drug names that look or sound alike. One potential method of reducing these errors is to highlight sections of names on labels in order to emphasize the differences between similar products. METHOD: In Experiments 1 and 2, participants were timed as they decided whether similar name pairs were the same name or two different names. Experiment 3 was a recognition memory task. RESULTS: Results from Experiments 1 and 2 showed that highlighting sections of words using tall man lettering can make similar names easier to distinguish if participants are aware that this is the purpose of the intervention. Results from Experiment 3 suggested that tall man lettering and/or color does not make names less confusable in memory but that tall man letters may increase attention. CONCLUSION: These findings offer some support for the use of tall man letters in order to reduce errors caused by confusion between drug products with look-alike names. APPLICATION: The use of tall man letters could be applied in a variety of visual presentations of drug names--for example, by manufacturers on packaging, labeling, and computer software, and in pharmacies on shelf labels. Additionally, this paper demonstrates two meaningful behavioral measures that can be used during product design to objectively assess confusability of packaging and labeling. PMID- 16696256 TI - Multiple user considerations and their implications in medical error reporting system design. AB - OBJECTIVE: The study examined the differences between physicians and clinical assistants in their preferences for a statewide medical error reporting system. BACKGROUND: Medical error reporting systems have been proposed as a means for studying the causes of medical error. Knowledge of user similarities and differences is needed for the development of design guidelines for medical error reporting systems. METHOD: Separate focus groups composed of 8 physicians and 6 clinical assistants (physician clinical support staff) were conducted. One-hour focus group meetings were conducted via toll-free teleconference lines one to two times per month for 9 months. All conversations were audiotaped and transcribed for analysis. An inductive content analysis was conducted. RESULTS: Eighty-six major and minor themes emerged. Differences between physicians and clinical assistants included rules and regulations governing the use of the system, the medium of reporting, and aspects of the organization that may affect reporting levels. CONCLUSIONS: Although physicians and clinical assistants shared similar preferences and beliefs surrounding error reporting, there were differences that need to be considered if medical error reporting systems are to be effective. APPLICATION: To successfully deploy a medical error reporting system, the system itself must be designed for the potential users. This study uncovered previously underappreciated issues that should be incorporated into the design and implementation process. Actual or potential applications of this research include the improvement of the design and implementation of medical error reporting systems to account for the needs of different types of users. PMID- 16696257 TI - The right picture is worth a thousand numbers: data displays in anesthesia. AB - OBJECTIVE: To review the literature on data displays in anesthesia identifying issues and developing design recommendations. BACKGROUND: Unexpected incidents are common in critical care medicine. Adverse outcomes are frequently the catastrophic endpoints of an "evolving" chain of subtle incidents. One strategy to reduce the likelihood of an adverse patient outcome during anesthesia is to improve the anesthesiologist's ability to detect, diagnose, and treat critical incidents. METHOD: A literature review and analysis of data displays. RESULTS: Current numerical and waveform displays do not support anesthesiologists optimally. An alternative is graphical displays that functionally integrate variables into objects. In a well-designed graphic object, deviations from normal are shown by distortions in the object's symmetry. The emerging patterns that result from distorted symmetry facilitate the correct diagnosis. When treatment is effective, an object's shape is restored to normal. Graphical displays can be an effective tool in supporting anesthesiologists' situation awareness. CONCLUSION: Problems related to graphical displays have delayed their use in anesthesia, including the lack of conclusive clinical evidence of their value. However, currently more evidence is accumulating that graphical displays have the potential to improve clinical performance. The successful development of these graphical displays takes into account task requirements, a user's perceptual processes, and task-specific cognition. APPLICATION: This paper provides suggestions for the development of more effective displays in anesthesiology. Graphical displays can increase the anesthesiologist's situation awareness and improve clinical performance. Clinical use of these displays has the potential to significantly improve patient safety. PMID- 16696258 TI - The efficacy of a medical virtual reality simulator for training phlebotomy. AB - OBJECTIVE: The present study compared the effectiveness of a virtual reality (VR) simulator for training phlebotomy with that of a more traditional approach using simulated limbs. BACKGROUND: Phlebotomy, or drawing blood, is one of the most common medical procedures; yet, there are no universal standards for training and assessing performance. The absence of any standards can lead to injuries and inaccurate test results if the procedure is improperly performed. METHOD: Twenty 3rd-year medical students were trained under one of the two methods and had their performance assessed with a 28-item checklist. RESULTS: The results showed that performance improvements were limited to those who trained with the simulated limbs, and a detailed comparison of the two systems revealed several functional and physical differences that may explain these findings. CONCLUSION: Participants trained with simulated limbs performed better than those trained with a VR simulator; however, the metrics recorded by the VR system may address some aspects of performance that could eventually prove beneficial. APPLICATION: The present study highlights the potential for medical simulators to improve patient safety by enabling trainees to practice procedures on devices instead of patients. Applications of this research include training, performance assessment, and design of simulator systems. PMID- 16696259 TI - Drug delivery as control task: improving performance in a common anesthetic task. AB - OBJECTIVE: To conceptualize delivery of anesthesia as a control task, similar to control tasks in nonmedical domains, and to evaluate how presentation of new information and feedback affects task performance. BACKGROUND: In anesthesia, integrated monitors that show intravenous drug and effect-site concentrations in a patient currently do not exist. However, using real-time displays of intravenous anesthetic concentrations and effects could significantly enhance intraoperative clinical performance. Pharmacological models are available to estimate past, present, and future drug concentrations in the brain and to predict the drug's physiological effects. A display that integrates pharmacological models and visualizes drug concentrations was developed and tested to see if this drug display significantly improved clinical performance. METHOD: Thirty-three anesthesiologists with different levels of expertise administered anesthesia to simulated patients in a high-fidelity patient simulator. The experimental group used a drug display that visualized drug concentrations in real time, whereas the control group administered drugs without this information. RESULTS: Anesthesiologists using the drug display achieved better hemodynamic control of the simulated patient than did the control group. Similarly, the drug display enabled anesthesiologists to wake up and reanimate the patient faster. CONCLUSION: Visual feedback of drug concentrations leads to superior performance in the delivery of anesthesia. Drug delivery can be conceptualized within a control theoretical framework. Finally, the drug display has significant clinical potential to increase patient safety. APPLICATION: Clinical performance in delivering anesthesia depends on feedback. By providing this feedback, the drug display supports clinicians' ability to more precisely and safely administer anesthesia. PMID- 16696260 TI - The potential of training to increase acceptance and use of computerized decision support systems for medical diagnosis. AB - OBJECTIVE: The goals of this study were to understand the reasons underlying the limited use of medical decision-support tools and to explore the potential of a computer-based tutorial to mitigate barriers to use. BACKGROUND: Medical decision support tools such the Acute Cardiac Ischemia Time-Insensitive Predictive Instrument (ACI-TIPI) have demonstrated statistical validity and clinical impact for patient safety but have seen limited adoption and use. METHODS: The study developed a brief Web-based "demystifying" ACI-TIPI tutorial employing case-based training and evaluated the effectiveness of that tutorial in changing self reported attitudes and behaviors. RESULTS: Clinicians using the tutorial reported greater understanding of how to use the ACI-TIPI score appropriately and increased confidence in the score. Case studies in the tutorial that provided examples of how to use the score for actual cases were rated as especially helpful. CONCLUSION: This study suggests that a primary barrier to the use of statistical decision support tools for patient diagnosis is lack of training or experience in combining a population-based numerical risk score with other diagnostic information about the individual patient's case that is not considered in that score. The results of this study indicate that there is a potential for a relatively brief tutorial to increase acceptance and use of decision support tools for medical diagnosis. APPLICATION: These findings have the potential for the identification of methods to help clinicians learn how to use statistical and probabilistic information to better assess risk and to promote integration of decision support tools into medical decision making for improvement of patient safety. PMID- 16696261 TI - +Gz acceleration loss of consciousness: time course of performance deficits with repeated experience. AB - OBJECTIVES: We examine the time course of performance recovery from gravity induced loss of consciousness (GLOC) and evaluate the utility of exposing participants to repeated bouts of GLOC in promoting recovery time. BACKGROUND: A substantial number of accidents among fighter pilots have resulted from episodes of GLOC. U.S. Air Force doctrine holds that when pilots experience GLOC, impairment lasts for 24 s, in which there are 12 s of complete unconsciousness and 12 s of confusion. However, there is reason to suspect that performance efficiency associated with GLOC is degraded well before unconsciousness sets in and that more than 24 s are required for performance efficiency to return to baseline levels. Additionally, there is a belief that repeated exposure to GLOC will reduce recovery time. METHOD: Centrifuge simulators were used to induce GLOC in U.S. Air Force personnel with math and tracking tasks employed to emulate flight performance. Participants were tested once per week for 4 consecutive weeks. RESULTS: On average, performance deficits appeared 7.44 s prior to the onset of unconsciousness and persisted for 55.6 s following the GLOC event. Repeated exposure failed to moderate these results. CONCLUSION: The temporal course of performance deficits produced by GLOC far exceeds prior estimates. The problem is more serious than previously envisioned and it is not alleviated by repeated exposure to GLOC. APPLICATION: U.S. Air Force doctrine regarding the severity of GLOC and the utility of repeated exposure to this problem needs to be revised and these data incorporated into future aircraft auto recovery systems. PMID- 16696262 TI - Keyswitch orientation can reduce finger joint torques during tapping on a computer keyswitch. AB - OBJECTIVE: To examine the effects of keyswitch orientation on joint torques. BACKGROUND: The fingertip produces primarily vertical forces during single-finger tapping on a computer keyswitch. However, horizontal force components within the sagittal plane of the finger could reduce net joint torques. METHOD: Eleven participants tapped on a keyswitch oriented in three directions: vertical, tilted 30 degrees such that when pressed it moved away from the user (similar to a positive-tilt keyboard), and tilted 30 degrees such that when pressed it moved toward the user (similar to a negative-tilt keyboard). Participants also tapped on a prototype cantilever keyswitch design in which the key cap moves along the arc of a bending beam gradually away from the user. Miniature electro-optical goniometers measured the finger posture, and a two-axis force sensor measured fingertip forces. RESULTS: Tapping on a keyswitch oriented such that it moves away from the user when pressed reduced net joint torques by 47% relative to tapping on a vertically orientated keyswitch and by 56% relative to tapping on a keyswitch oriented toward the user, whereas the cantilever design resulted in 14% decreases in net joint torque relative to the vertical orientation. CONCLUSION: Reductions of torques resulted from decreasing the moment arm of the fingertip force about the joints. APPLICATION: Keyboard design should incorporate keyswitch mechanism angles along with other postural and geometric constraints to reduce exposure of the finger joints and muscles to force during typing. PMID- 16696263 TI - Forearm electromyographic changes with the use of a haptic force-feedback computer mouse. AB - OBJECTIVE: To examine changes in biomechanical and motor control associated with a force-feedback computer mouse. BACKGROUND: Haptic computer mice can improve the movement times for point-and-click tasks; however, changes in upper extremity biomechanics and motor control are unknown. METHOD: Eighteen people (ages 22-37 years) performed a point-and-click task 80 times using a force-feedback computer mouse across three different conditions: (a) no force feedback, emulating a conventional mouse; (b) a single attractive force field at the desired target that pulls the mouse to the center of the target; and (c) an attractive force field at the desired target as well as others between the two possible targets, distracting the user from the intended target. Cursor kinematics, wrist posture, and electromyographic (EMG) forearm muscle activity were recorded. RESULTS: The point-and-click movements were 30% faster with the addition of a single force field and 3% faster with the addition of multiple force fields. The Fitts' law index of performance metrics improved from 2.9 bits/response to 4.1 bits/response for multiple attractive fields and to 6.0 bits/response for a single force field. For the distracting force fields, the cursor maximum velocities were over 50% faster. EMG amplitude values were largest for the distracting force fields. These data suggest that the operator uses increased muscle activity to accelerate the mouse through the distracting force fields. CONCLUSION: When implementing attractive haptic force fields, one needs to consider how to reduce these observed effects of potential distracting force fields. APPLICATION: Applications include human-computer interface design for pointing devices extensively used for the graphical user interface. PMID- 16696264 TI - Stability ball versus office chair: comparison of muscle activation and lumbar spine posture during prolonged sitting. AB - OBJECTIVE: The objective of the study was to evaluate the differences between sitting on a stability ball and in an office chair in terms of trunk muscle activation and lumbar spine posture. BACKGROUND: Stability balls have become increasingly popular as an alternative to office chairs to help reduce the prevalence of low back pain; however, little research has been conducted on their use as office chairs. METHODS: The 14 participants (7 men, 7 women) were required to sit on both a stability ball and an office chair for 1 hour each while performing various computer workstation tasks throughout the sitting periods. The activation of eight muscles and lumbar spine posture were measured and analyzed. RESULTS: Increased muscle activation in thoracic erector spinae (p = .0352), decreased pelvic tilt (p = .0114), and increased perceived discomfort (p < .0001) while sitting on the stability ball were observed. CONCLUSIONS: The small changes in biological responses when sitting on a stability ball as compared with an office chair, combined with the increased reported discomfort while on the ball, suggests its use for prolonged sitting may not be advantageous. APPLICATION: Prolonged sitting on a stability ball does not greatly alter the manner in which an individual sits, yet it appears to increase the level of discomfort. Therefore, it is important to fully explore a new chair design and consult scientific research before implementing its use. PMID- 16696265 TI - Spatial ability subfactors and their influences on a computer-based information search task. AB - OBJECTIVE: The present study examined the relationship between two distinct subfactors of spatial ability and performance in an information search task modeled on browsing the Web. BACKGROUND: Previous studies have found relationships between various measures of spatial ability and performance in a wide variety of computer-based tasks. METHOD: In the search task 101 participants (18-29 years of age) searched for the answer to a question by navigating the system. They completed the experimental task as well as a battery of cognitive ability measures that included two different measures of spatial ability. RESULTS: The results indicate that spatial orientation ability was related to performance with tasks that were high in their navigational requirement (engendered by the use of a novel aid), whereas spatial visualization was unrelated to performance in any task condition. CONCLUSION: A closer inspection of the cognitive requirements of a task may reveal what interventions could be most useful when designing computer systems or developing training programs. APPLICATION: Given the unique differences between the different spatial abilities, the current results suggest the design of navigational aids that place less demand on spatial orientation ability. PMID- 16696266 TI - Pilots strategically compensate for display enlargements in surveillance and flight control tasks. AB - OBJECTIVE: Experiments were conducted to assess the impact of display size on flight control, airspace surveillance, and goal-directed target search. BACKGROUND: Research of 3-D displays has shown that display scale compression influences the perception of flight path deviation, though less is known about the causes that drive this effect. In addition, research on attention-based tasks has shown that information displaced to significant eccentricities can amplify effort, but it is unclear whether the effect generates a performance difference in complex displays. METHOD: In Experiment 1, 16 pilots completed a low-fidelity flight control task under single- and dual-axis control. In Experiment 2, the control task from Experiment 1 was scaled up to a more realistic flight environment, and pilots performed hazard surveillance and target search tasks. RESULTS: For flight control, pilots exhibited less path error and greater stick activity with a large display, which was attributed both to greater enhanced resolution and to the fact that larger depictions of error lead to greater urgency in correcting deviations. Size did not affect hazard surveillance or search, as pilots were adaptive in altering scanning patterns in response to the enlargement of the displays. CONCLUSION: Although pilots were adaptive to display changes in search and surveillance, display size reduction diminished estimates of flight path deviation and control performance because of lowered resolution and control urgency. APPLICATION: Care should be taken when manipulating display size, as size reduction can diminish control performance. PMID- 16696267 TI - Ergonomic guidelines for traffic sign design increase sign comprehension. AB - OBJECTIVE: This research directly tests the relationship between comprehension probability of highway signs and the extent to which they comply with three ergonomic principles of design: sign-content compatibility, familiarity, and standardization. BACKGROUND: A recent study that evaluated comprehension of traffic signs in four different countries showed that comprehension level varies widely and is apparently related to the extent that the sign's design incorporates ergonomic guidelines for good design (D. Shinar, R. E. Dewar, H. Summala, and L. Zakowska 2003). METHOD: Participants were presented with 30 signs and asked to describe each sign's meaning. They then evaluated each sign in terms of each of three ergonomic principles. In addition, a group of human factors/ergonomics experts evaluated each sign on its standardization and compatibility. RESULTS: There were high correlations between the ratings of the students and the ratings of the experts on compatibility (rho = .94) and on standardization (rho = .84), validating the use of the students' evaluations of the signs on these variables. There was a great variability in signs' comprehension and high and statistically significant correlations between the comprehension level of each sign and the extent to which it complied with compatibility (R = .76), familiarity (R = .89), and standardization (R = .88) principles. CONCLUSIONS: The more signs conform to universal ergonomic principles of good design, the more likely they are to be comprehended by drivers of different cultural backgrounds. APPLICATION: Sign design should be guided by established ergonomics principles to enhance comprehension, especially for drivers who have not had any prior encounters with specific signs. PMID- 16696268 TI - Examining the impact of cell phone conversations on driving using meta-analytic techniques. AB - OBJECTIVE: The performance costs associated with cell phone use while driving were assessed meta-analytically using standardized measures of effect size along five dimensions. BACKGROUND: There have been many studies on the impact of cell phone use on driving, showing some mixed findings. METHODS: Twenty-three studies (contributing 47 analysis entries) met the appropriate conditions for the meta analysis. The statistical results from each of these studies were converted into effect sizes and combined in the meta-analysis. RESULTS: Overall, there were clear costs to driving performance when drivers were engaged in cell phone conversations. However, subsequent analyses indicated that these costs were borne primarily by reaction time tasks, with far smaller costs associated with tracking (lane-keeping) performance. Hands-free and handheld phones revealed similar patterns of results for both measures of performance. Conversation tasks tended to show greater costs than did information-processing tasks (e.g., word games). There was a similar pattern of results for passenger and remote (cell phone) conversations. Finally, there were some small differences between simulator and field studies, though both exhibited costs in performance for cell phone use. CONCLUSION: We suggest that (a) there are significant costs to driver reactions to external hazards or events associated with cell phone use, (b) hands-free cell phones do not eliminate or substantially reduce these costs, and (c) different research methodologies or performance measures may underestimate these costs. APPLICATION: Potential applications of this research include the assessment of performance costs attributable to different types of cell phones, cell phone conversations, experimental measures, or methodologies. PMID- 16696269 TI - Impact of P4P. PMID- 16696271 TI - Important phlebotomy techniques. PMID- 16696270 TI - On cord blood labeling. PMID- 16696272 TI - CLS: how did we get here? PMID- 16696273 TI - The laboratory's role in the universal electronic health record. PMID- 16696274 TI - Under the blue top: part II fine-tune your ISIs. PMID- 16696275 TI - A world-class accommodates worldwide service. Armed Forces Institute of Pathology Lab is chosen as MLO's 2006 winner. PMID- 16696276 TI - Ron Andrews on Clarient's collaboration. PMID- 16696277 TI - Faulty time clock prompts tardiness policy review. PMID- 16696278 TI - Anastomotic leak after esophagectomy. AB - Anastomotic leaks continue to be a source of considerable morbidity and mortality after esophageal resection. Careful attention to the contributing factors to the development of a leak, particularly conduit ischemia and anastomotic technique, can reduce the incidence of anastomotic complications postoperatively. Selective management of leaks, including non-operative treatment when indicated, will maximize the chance for a successful outcome. PMID- 16696279 TI - Esophageal conduit necrosis. AB - A cumulative review of the prevalence of esophageal conduit necrosis is summarized in Table 4. The spectrum of conduit ischemia is broad and includes cases in which there is anastomotic leak or stricture as well as cases in which there is frank graft necrosis. Many of the studies that the authors reviewed do not specify the exact nature of postoperative ischemic complications or how they are defined. Therefore, postoperative conduit ischemia is reported globally. Based on the authors' review, average rates of ischemic complications for stomach, colon, and jejunum are 3.2%, 5.1%, and 4.2%, respectively. Results for colon and jejunum include results for both long- and short-segment grafting. Most reports that compare outcomes using different esophageal conduits demonstrate findings similar to the authors'. Davis and colleagues compared results with colon versus gastric conduit esophageal reconstruction. They found that operative mortality, anastomotic leaks, and conduit ischemia rates were all lower for the stomach than for the colon. Specifically, ischemia of the stomach conduit was 0.5%, compared with 2.4% for the colon conduit. Moorehead and Wong, in a large series of 760 esophagectomy patients in whom the stomach, colon, or jejunum was used for reconstruction, demonstrated that the stomach had the lowest incidence of conduit ischemia (1%), followed by jejunum (11.3%), then colon (13.3%). Some of the factors they identified as correlating with the risk of ischemia include length of conduit, technique of stomach graft preparation, whether anastomosis is in the neck or chest, and route of passage of the conduit. Mansour and colleagues compared their results using bowel interposition (either colon or jejunum) to reconstruct the resected esophagus. The authors report an overall mortality of 5.9%, and 3% conduit ischemia. All ischemia was noted in the colon conduits harvested from the left side. No ischemic complications were noted from jejunal segments. Briel and colleagues compared stomach versus colon conduit use after esophagectomy. They note an overall incidence of ischemia of 9.2%. In their series, the incidence of ischemia for stomach and colon was 10.4% and 7.4%, respectively. Anastomotic leak and stricture rates, both thought to be sequelae of ischemia, also were lower for colon conduit use than for stomach conduit. Multivariate analysis identified patient comorbidities as the only independent risk factor for conduit ischemia. The authors use their findings to support the preferential use of colon conduits rather than stomach conduits. The incidence of colon conduit ischemia (7.4%) is directly in line with all other published results, including the cumulative review by the authors of this article, whereas the rate of stomach conduit ischemia (10.4%) is considerable higher than in most other studies. Esophageal conduit necrosis is an uncommon but disastrous complication of esophageal surgery. Careful selection of patients for surgery, preoperative evaluation of the proposed conduit, and meticulous operative technique are the best defenses against conduit ischemia. Postoperatively, surgeons should have a high index of suspicion for this complication. Unexplained tachycardia, respiratory failure, leukocytosis, or any evidence for graft or anastomotic leak should prompt a search for conduit ischemia. The diagnosis is made by contrast esophagography, endoscopy, or direct operative inspection. There is no documented salvage technique once ischemia is identified. Treatment for mild cases may be supportive, with or without management of anastomotic leak. More severe cases of necrosis require debridement and conduit take-down with proximal esophageal diversion and placement of enteral feeding tubes. Reconstruction can be planned for later if possible. The majority of the data demonstrates that risk of ischemia is related to conduit type, length of conduit, comorbidities, and operative technique. The stomach has the lowest reported incidence of conduit ischemia, followed by the jejunum, and colon. In the future, methods to predict conduit ischemia more accurately at the time of surgery may further reduce the incidence of this disastrous complication. PMID- 16696280 TI - Recurrent laryngeal nerve injuries after esophagectomy. AB - Unilateral vocal-cord paralysis after esophagectomy involving a cervical anastomosis is uncommon in experienced hands but can cause marked morbidity because of pulmonary complications. It is important for the surgeon to understand the anatomy of the recurrent laryngeal nerve to avoid this complication. The diagnosis is usually easily made at the bedside and can be confirmed by visualization of the glottis. Early treatment may include supportive care with a speech pathologist or injection of the vocal-cord with temporary material to improve glottic closure. Definitive treatment can best be performed by medialization thyroplasty. PMID- 16696281 TI - Respiratory complications after esophagectomy. AB - Pulmonary complications are the major source of morbidity and mortality after esophageal resection, and numerous studies have identified various associated with these complications. This article discusses preoperative, intraoperative, and postoperative factors affecting pulmonary complications and strategies to reduce these complications after esophagectomy. PMID- 16696282 TI - Chylothorax after esophagogastrectomy. AB - In conclusion, chylothorax after esophagectomy is a devastating complication with high mortality rates if not corrected. A heightened awareness of this complication with early diagnosis and aggressive reoperation leads to excellent outcome. Reoperation is not indicated only when medical therapy significantly slows the daily loss of chyle and there are no metabolic consequences. The early decision to reoperate avoids the high morbidity of a persistently unchecked chylothorax. Reoperation should be based on the approach initially used for the esophagectomy, the location of the leak, and the side that has the chylothorax. The conduit should be handled carefully at the time of reoperation, the leak identified, the duct or the leaking nodal basin clipped and glued, and a pleurodesis performed. Following these principles minimizes the morbidity of a serious postoperative complication. PMID- 16696283 TI - Functional conduit disorders after esophagectomy. AB - Unfortunately normal gastrointestinal function after an esophagectomy is rare. Most patients will never eat the way they did before their illness. Most patients require smaller more frequent meals. It is common for patients to loose up to 15% of their body weight from the time of diagnosis through the first 6 months postoperatively, but fortunately this trend levels off after 6 months. Dumping syndrome, delayed gastric emptying, reflux, and dysphagia can all contribute to nutritional deficiency and poor quality of life. There is no one surgical modification to eliminate any one of these complications, but several guidelines can help reduce conduit dysfunction. Most patients seem to benefit from a 5-cm wide greater-curvature gastric tube brought up through the posterior mediastinum. The gastric-esophageal anastomosis should be placed higher than the level of the azygous vein. Drainage procedures seem to be helpful, especially when using the whole stomach as a conduit. Early erythromycin therapy significantly aids in the function of the gastric conduit. Proton-pump inhibitors are important for improvement of postoperative reflux symptoms and to help prevent Barrett's metaplasia in the esophageal remnant. Single-layer hand-sewn or semi-mechanical anastomoses provide greater cross-sectional area and fewer problems with stricture. When benign strictures occur, early endoscopy and dilation with proton pump inhibition greatly reduces the morbidity. Patients should be instructed to eat six small meals a day and to remain upright for as long as possible after eating. Simple sugars and fluid at mealtime should be avoided until the function of the conduit is established. PMID- 16696284 TI - Anastomotic stricture complicating esophagectomy. AB - Regardless of the definition, anastomotic strictures are a common complication after esophagectomy and adversely affect quality of life. They are best avoided by careful surgical technique that minimizes conduit ischemia during preparation, placement, and anastomosis. Anastomotic technique must assure an adequate anastomotic area. The Collard anastomosis, a significant advance in the construction of esophagogastric anastomoses, routinely assures adequate anastomotic area and thus assures fewer anastomotic strictures. The use of small diameter (21-mm and 25-mm) circular staplers is discouraged, because they are unquestionably associated with the occurrence of major anastomotic strictures. Anastomotic leaks precede many anastomotic strictures, but strictures are not inevitable after leaks. Other variables are less reliably associated with anastomotic strictures. Treatment requires diagnosis and exclusion of recurrent cancer and other causes of stricture. Dilation is safe, but diligence with repeated sessions is necessary to restore swallowing. Reoperation is rarely required. PMID- 16696285 TI - The effects of perioperative blood transfusion on morbidity and mortality after esophagectomy. AB - The effect of blood transfusion on outcomes in esophageal surgery remains controversial. The contrasting conclusions drawn from a number of retrospective analyses with different methodologies create a landscape that is difficult to interpret. Because of the scope of esophageal resection, the need for blood transfusion cannot be eliminated. What recommendations then, if any, can be made for the practicing surgeon? First, surgeons and anesthesiologists need to reevaluate their transfusion thresholds. The age-old practice of keeping the hemoglobin above 10 g/dL has very little evidence-based support. A multicenter, randomized, controlled clinical trial in Canada demonstrated that a restrictive strategy of blood transfusion, in which patients were transfused only for a hemoglobin level of less than 7 g/dL, was at least as effective as and possibly was superior to a liberal transfusion strategy in critically ill patients. It has also been estimated that more than 25% of patients undergoing colorectal resections may receive at least one unit of unnecessary blood. Further, the immediate reduction in the hemoglobin concentration caused by the normovolemic hemodilution associated with surgery and crystalloid fluid replacement is not associated with any increased morbidity or mortality. If these data are examined in the context of the results of Langley and Tachibana indicating that a threshold amount of blood needs to be transfused to impact outcomes, it becomes even more important to limit transfusion to only the amount that is essential. Thus, surgeons and anesthesiologists should adopt a more stringent set of requirements for blood transfusion. Second, with the proven feasibility and reduction in infectious complications associated with autologous blood-donation programs, any patient who meets the criteria discussed here should be encouraged to participate in such a program. Although the effect of autologous blood on cancer outcomes remains unclear, the other advantages certainly make such a program worthy of consideration. This discussion leads to a final point, namely that patients should be encouraged, whenever possible, to participate in clinical trial research. The only way that the community of surgeons treating patients who have esophageal cancer can hope to address properly the question of how blood transfusion affects outcomes is with well-designed clinical trials. A large, multicenter, randomized trial (level I) would be ideal. Short of such a trial, inclusion criteria and study methodology should be discussed among various institutions to avoid the differences in studies that make direct comparisons of results among different investigators difficult and potentially meaningless. This measure would at least allow different level II to IV data to be compared directly with some validity. PMID- 16696286 TI - The volume-performance relationship in esophagectomy. AB - The current literature demonstrates that both hospital volume and surgeon volume are inversely related to operative mortality for esophageal resection. Given the heterogeneity of the volume-outcome literature, it is difficult to identify minimum volume thresholds at which satisfactory performance is achieved. Both volume-based hospital referral and process improvement provide compelling and sometimes competing strategies for translating evidence about volume-outcome relationships into policy. Each approach has significant limitations that preclude uniform implementation nationwide but nonetheless should be a focus for continued investigation and application, with the goal of providing uniform and high-quality care after esophagectomy. PMID- 16696287 TI - Complications of achalasia surgery. AB - The development of minimally invasive techniques have revolutionized the surgical therapy of achalasia and made myotomy with or without partial fundoplication the treatment of choice for most patients. Complications do occur, however, and every effort must be made to minimize their occurrence and identify and treat them effectively when they occur to maximize the likelihood of an excellent outcome in these patients who have a benign but incurable disorder. Further studies on the best location for the myotomy as well as the best type of fundoplication will help refine the procedure for future generations of patients. PMID- 16696288 TI - Complications after surgery for gastroesophageal reflux disease. AB - Most complications after surgery for GERD can be avoided by experience and proper surgical technique. Often, what is termed a "slipped" or "twisted" wrap is one that was not properly constructed during the initial surgery. These technical errors can be avoided by complete mobilization of the stomach and esophagus, removal of the epigastric fat pad to identify esophageal shortening, and preservation of both vagus nerves. It is critical to avoid these errors, because an improperly constructed wrap will probably condemn the patient to significant dysphagia, recurrent reflux, and the need for reoperation. Should reoperation be required, the wrap should be completely dismantled so the technical error can be identified and a proper antireflux mechanism created. PMID- 16696289 TI - Opportunities and responsibilities for nurses in the genomic era. PMID- 16696290 TI - Introduction to genetics for otorhinolaryngology nurses. AB - Advances in human genetics have led to considerable changes in the way health professionals, especially nurses, understand and care for their patients. This article is focused on the clinical implications of genetic aspects of conditions of the head and neck, written for otorhinolaryngology (ORL) and head-neck nurses, but relevant for other nurses as well. This article presents information about basic inheritance patterns (autosomal dominant and recessive, X-linked, and mitochondrial) and compares single-gene disorders to complex or mutifactorial diseases. Cleft lip and palate disorders (Robin sequence and Van der Woude syndrome) and other craniofacial conditions (craniosynostosis, Treacher-Collins syndrome, and velo-cardio-facial syndrome) are discussed along with non-syndromic and syndromic hearing loss. Discussion of the psychosocial aspects of genetic conditions of the head and neck and a glossary of genetic terms are included. PMID- 16696291 TI - Factors associated with adult poisoning in northern Malaysia: a case-control study. AB - Data on adult risk factors associated with drug or chemical poisonings in Malaysia are scarce. The objective of the study was to identify possible risk factors associated with adult admissions to the Penang General Hospital (PGH) due to chemical poisoning and/or drug overdose. The present study was a case-control study, conducted over 18 weeks. One hundred acutely poisoned adult patients admitted to PGH during the period from September 2003 to February 2004 were considered as cases. Two hundred patients admitted to the same medical wards for other illnesses, during the same period, were matched for age and gender with the poisoned cases and thus selected as controls. McNemar test and binary logistic were used for univariate analysis and logistic regression analysis for multivariate analyses. The odds ratio (OR) and its 95% confidence interval (95% CI) were calculated for each predictor variable. Positive histories of psychiatric illness and previous poisoning, problems in boy/girl friend relationships, family problems, marital problems, Indian ethnicity, Chinese ethnicity, living in rented houses and living in a household with less than five people were significant risk factors associated with adult admissions due to poisoning. PMID- 16696292 TI - Calcium gluconate and sodium succinate for therapy of sodium fluoroacetate experimental intoxication in cats: clinical and electrocardiographic evaluation. AB - Sodium fluoroacetate (SFAC) or Compound 1080 is a potent rodenticide, largely used after 1946 for rodent and home pest control. The toxic effects of SFAC are caused by fluorocitrate action, a toxic metabolite, which has a competitive action with aconitase enzyme, leading to citrate accumulation and resulting in interference in energy production by Krebs cycle blockade. In the present study, domestic cats were intoxicated with oral doses of fluoroacetate (0.45 mg/kg). The intoxicated animals presented emesis, diarrhea with abdominal pain posture and an abdominal palpation, tachypnea, bilateral midriasis, hypothermia, hyperexcitability and convulsions. Blood gas analysis indicated decreased pH and bicarbonate levels. Serum ionized calcium was also decreased. ECG showed non specific changes in ventricular repolarization and ventricular arrhythmias. The survival rate was 75% in the treated group with calcium gluconate and sodium succinate and 37.5% in the nontreated group. PMID- 16696293 TI - Epidemiology of jellyfish stings reported to poison centers in Texas. AB - This study examined the relationship between selected factors and all human exposures involving jellyfish stings reported to Texas poison centers. Cases were obtained retrospectively from calls to poison centers in Texas and included all reported human exposures during 1998-2004 involving jellyfish stings. The distribution of cases was determined for a variety of demographic and clinical parameters. There were 423 total cases. Among the cases with a known patient age, 19.8% were <6 years of age, 53.5% were age 6-19 years, and 26.7% were >19 years of age. Males accounted for 52% of the cases. Of the 118 cases with a known clinical outcome, 0.8% had no effect, 80.5% had minor effects, and 18.6% had moderate effects. Counties along the Gulf Coast accounted for 72.3% of the calls. This information can be used to identify those portions of the population most at need of education regarding the prevention and treatment of jellyfish stings. PMID- 16696294 TI - Neuroprotective effect of Acorus calamus against middle cerebral artery occlusion induced ischaemia in rat. AB - The neuroprotective potential of ethanol:water (1:1) extract of rhizomes of Acorus calamus (AC-002) has been investigated in middle cerebral artery occlusion (MCAO)-induced ischaemia in rats. A significant behavioural impairment in Rota Rod performance and grid walking was observed in rats, 72 hours after MCAO as compared to sham-operated animals. These rats also exhibited an increase in lipid peroxidation (cortex -157%, corpus striatum - 58%) and a decrease in glutathione levels (cortex - 59%, corpus striatum - 34%) and superoxide dismutase (SOD) activity (cortex - 64%, corpus striatum - 32%) as compared to sham-operated animals. Ischaemic rats treated with AC-002 (25 mg/kg, p.o.) exhibited a significant improvement in neurobehavioural performance viz. Rota-Rod performance and grid walking as compared to the MCAO group. Interestingly, treatment with AC 002 in MCAO rats significantly decreased malonaldialdehyde levels in cortex as compared to ischaemic rats. A significant increase in reduced glutathione levels and SOD activity was also observed both in cortex and corpus striatum in MCAO rats treated with AC-002 in comparison to MCAO rats. Treatment with AC-002 in MCAO rats also reduced the contralateral cortical infarct area (19%) as compared to MCAO rats (33%). Neurological function score was improved in the AC-002 treated rats as compared to the MCAO group. The results of the present study indicate the neuroprotective efficacy of A. calamus in the rat model of ischaemia. PMID- 16696295 TI - Butanol ingestion in an airport hangar. AB - 1-Butanol is a colourless organic solvent with a rancid sweet odour. 1-Butanol ingestion may result in vomiting, abdominal pain, headache, drowsiness and unconsciousness. We present a 47-year-old male with no previous medical history, who was found comatose and soiled after having vomited while unconscious. On arrival, he had a Glasgow coma scale of 3, tachycardia, hypotension, shallow tachypnoic breathing, hypotonic muscles, absent myotatic reflexes and aromatic odour. The patient was intubated and treated with oxygen, dopamine and volume replacement therapy. Gastric lavage was performed and activated charcoal was given. His initial laboratory test revealed hypokaliemia, renal failure, acidosis with elevated lactate and hypercapnic respiratory insufficiency. Twelve hours after admission, the patient started to respond to a painful stimulus and 4 h later he was conscious. He was extubated 23 h after admission. All pathological laboratory results gradually returned within normal limits. The subsequent toxicological examination of gastric content and urine sample by gas chromatography revealed 1-butanol. On awakening, he confirmed ingestion of a solvent stored in an airport hangar. In conclusion, we describe a patient who ingested - a posteriori with suicidal intention - an unknown dose of 1-butanol. Symptoms were headache, vomiting, abdominal pain, coma, muscular hypotonus, hypotension, respiratory insufficiency and mixed acidosis. The patient totally recovered after supportive therapy over 30 h. In future cases, intravenous administration of ethanol or even hemodialysis can be considered analogous to the treatment of methanol and ethylene glycol poisoning. PMID- 16696296 TI - The influence of tobacco smoke and nicotine on antidepressant and memory improving effects of venlafaxine. AB - In experimental and clinical studies, central nicotinic systems have been shown to play an important role in cognitive function. Nicotinic acetylcholine receptors also mediate the reinforcing properties of nicotine (NIC) in tobacco products. A variety of studies have shown that acute treatment with NIC or nicotinic agonists can improve working memory function. Moreover, it is known that the monoaminergic system plays an important role in memory function. And there is evidence suggesting that prolonged use of NIC may exert antidepressant action via nicotinic receptors. The purpose of this study was to investigate the interactions between a novel antidepressant, venlafaxine (VEN), a blocker of noradrenaline and 5-hydroxytryptamine reuptake sites, and pure NIC in the context of antidepressant and memory function in tobacco smoke exposed and nonexposed rats. The animals were subjected to Porsolt's test for testing antidepressant activity and their memory function (spatial memory) was evaluated in the Morris Water Maze Test. In tobacco smoke non-exposed and exposed rats both single and chronic administration of VEN (20 mg/kg po) shortened immobility time. NIC (0.2 mg/kg sc) significantly reduced immobility time on the 1st, 7th and 14th test days in both non-exposed and exposed rats. Combined VEN+NIC treatment in tobacco smoke non-exposed rats reduced immobility too. This effect of the combination of drugs was significantly stronger as compared to the effects obtained after individual administration of VEN or NIC. In the group exposed to tobacco smoke, joint administration of VEN+ NIC induced a significant reduction of immobility as compared to the control and NIC groups. In the Morris Water Maze Test single and chronic administration of VEN, lower values of escape latencies and lower numbers of crossed quadrants were noted in both exposed and non-exposed rats, which indicates improved performance. After administering NIC we could observe improvement of spatial memory in both the exposed and non-exposed group. A similar effect of improvement of spatial memory was observed after joint administration of VEN and NIC. The study results support the involvement of nicotinic systems in memory processes in rats. Memory improvement and antidepressant effects following joint administration of VEN and NIC may depend on nicotinic interactions with monoaminergic systems and VEN may represent a new therapeutic approach to smoking cessation. PMID- 16696297 TI - Preliminary survey of ochratoxin A in millet, maize, rice and peanuts in Cote d'Ivoire from 1998 to 2002. AB - In a preliminary study, samples of millet (n =33) maize (n=41), rice (n=10) and peanuts (n=10) from Cote d'Ivoire were analysed for ochratoxin A (OTA) by HPLC with fluorimetric detection, followed by confirmation by cleavage of the OTA molecule using carboxypeptidase with HPLC separation and fluorimetric quantification of the released ochratoxin alpha (OTh). With the exception of four samples of peanuts, all samples showed OTA contamination, ranging from 3 to 1738 microg/kg. All cereals were contaminated and the OTA concentrations were in the range of 17-204 microg/kg for millet, 3-1738 microg/kg for maize, 9-92 microg/kg for rice and 0.6-64 microg/kg for peanuts, depending on the year of harvest. Most of the samples would not be accepted according to the EU regulatory limits for this mycotoxin. Following this survey, research for other mycotoxins and the evaluation of the exposure of the population is underway. PMID- 16696298 TI - Demographical, aetiological and clinical characteristics of poisonings in Mersin, Turkey. AB - BACKGROUND: As acute poisoning continues to be an important public health problem, and represents a frequent cause of admission in emergency departments, there is a constant need to have up to date information about it. The aim of this retrospective study was to evaluate the demographical, aetiological and clinical characteristics of acute poisonings who were admitted to a university hospital in the south part of Turkey. METHODS: The data of 195 acute poisoning patients who were admitted to the Emergency Department of Mersin University were retrospectively studied. RESULTS: Most poisonings occurred in the age range 0-6 years (57.5%) for children and 17-25 year (46.7%) for adults. The overall female to male ratio was 1.26. Drugs were the most common cause of poisonings (55.4%). Among the drug poisonings, psychotropics (41.7%) were the most frequent agent, followed by analgesics (20.4%). 67.7% of the cases presented to the emergency department within 3 hours of poisoning and the procedures for elimination of poison were performed in most of them. The clinical status of the patients on arrival was generally good. 69.2% of the patients were hospitalized. The mean duration of hospitalization was 2.4+/-2.8 days. 41.8% of the suicidal cases had a history of psychiatric disorders; 92.8% of the patients were discharged home after toxicological treatment; 2.6% of the cases died. CONCLUSION: Further studies are necessary to collect more data and these data will be helpful not only for determining risk factors of acute poisonings but also for developing education and prevention programmes. PMID- 16696299 TI - The influence of elastin-coated 520-nm- and 20-nm-diameter nanoparticles on human fibroblasts in vitro. AB - Magnetic nanoparticles have been used for biomedical purposes for several years. In recent years, nanotechnology has developed to a stage that makes it possible to engineer particles to provide opportunities for the site-specific delivery of drugs. To this end, a variety of iron oxide particles have been synthesized. The size and surface of the particles are crucial factors in the application of the particles. Therefore, this study involves the use of two types of magnetic nanoparticles derivatized with elastin and synthesized with differing diameters, compared with identical underivatized plain particles. This influence in vitro was assessed using human dermal fibroblasts and various techniques to observe cell-particle interaction, including light and fluorescence microscopy and scanning electron microscopy. The results indicate that derivatized particles induce alterations in cell behavior and morphology distinct from the plain particles, suggesting that cell response can be directed via specifically engineered particle surfaces. However, little difference was observed between the different diameters. PMID- 16696300 TI - Microscopical characterization of nanocapsules templated on ionic crystals and biological cells toward biomedical applications. AB - Nanocapsules, fuzzy assemblies of polyelectrolyte, represent a comparatively new class of colloids with controlled nanostructure and tunable properties. Due to the fact that the core as well as the dissolution influences the wall texture and the properties of the hollow capsules, the use of carbonate crystals as template is most convenient. Yeast cells constitute as a core candidate as well. They are a good system for testing the protective ability of shells and the permeability of the walls with respect to the needs of biological systems, namely, feeding and stability against attacks. The main features of the nanocapsules have been studied by two-photon, confocal, and atomic force microscopy. Nanocapsules are of biomedical interest because they can be used, for example, for the controlled release and targeting of drugs as well as for the protection of enzymes, proteins, and foreign cells. PMID- 16696301 TI - Inter-pulse interval between rectangular voltage pulses affects electroporation threshold of artificial lipid bilayers. AB - This paper describes experiments that determine how the inter-pulse interval between rectangular pulses in a train of pulses alters the threshold of electroporation of 1-pamitoyl 2-oleoyl phosphatidycholine bilayer lipid membranes. The bilayers were exposed to a train of sixteen 100-micros duration pulses. Threshold voltage and the sequence number of the pulse in the train, where onset of the electroporation occurred, were recorded for six inter-pulse intervals (infinity, 1000 micros, 100 micros, 10 micros, 1 micros, 0 micros). The threshold voltage of the pulse train decreased linearly with the logarithm of the inter-pulse interval. When the inter-pulse interval was 1 microm, electroporation threshold dropped to that of a single pulse with duration 1600 micros (equal to the sum of all pulse durations in the train). In this case, the occurrence of bilayer rupture was almost equally frequent for all pulses in the train. When the inter-pulse interval between the pulses exceeded 1 micros, the influence of the previous pulse on the response to the following pulse declined. It became more likely that the bilayer ruptured during the first half of the train. These experimental observations suggest that a train of pulses applied with short inter pulse interval (less than 1 ms) can lower the electroporation threshold of bilayer lipid membranes. PMID- 16696302 TI - Numerical simulation of protein stamping process driven by capillary force. AB - Numerical simulations based on first-principle conservation laws of mass and momentum are performed to observe flow characteristics during the stamping process. The protein solution is transferred by a new design of microstamps with microchannels and printed on a bottom substrate. Furthermore, key physics of the stamping process and the control factors achieving uniform spot size can be identified and optimized after these simulations. PMID- 16696303 TI - Opening eyes, opening minds. The Office of Minority Affairs reaches out to a new generation of students. PMID- 16696304 TI - The knight in the laboratory. Battling the side effects of radiotherapy through bone regeneration. PMID- 16696305 TI - A Sherlock Holmes of dental medicine. Following a new trail of clues to crack the mystery of oral cancer. PMID- 16696306 TI - Role of nitiic oxide and nitric oxide synthases in ischemia-reperfusion injury in rat organotypic hippocampus slice. AB - To investigate the effects of ischemia-reperfusion on the levels of nitric oxide and nitric oxide synthase isoforms (nNOS and iNOS), rat organotypic hippocampus slice were cultured in vitro and subjected to ischemia by oxygen-glucose deprivation (OGD) for 30 min and then placed in the normal culture condition. The ischemia-reperfusion produced a time-dependent increase in nitrite levels in the culture medium. Reverse transcriptional-polymerase chain reaction showed augmented levels of mRNA for both nNOS and iNOS when compared with control at 12 h and remained increase at 36 h after OGD (P < 0.05). The protein levels of both nitric oxide synthase isoforms increased significantly as determined by Western Blot. OGD also caused neurotoxicity in this model as revealed by the elevated lactate dehydrogenase (LDH) efflux into the incubation solution. The results suggest that organotypic hippocampus slice is a useful model in studying ischemia reperfusion brain injury. NO and NOS may play a critical role in the ischemia reperfusion brain damage in vitro. PMID- 16696307 TI - P53 gene mutation and expression of MDM2, P53, P16 protein and their relationship in human glioma. AB - To investigate the effect of P53 protein accumulation and p53 gene mutation in the pathogenesis of glioma and to study the role of MDM2, P53 and P16 protein in glioma formation and progression and their relationship with each other, LSAB immunohistochemical staining method and non-isotopic PCR-SSCP techniques were used to detect the expression of MDM2, P53 and P16 protein and p53 gene mutation in 48 cases of gliomas. The results showed that the positive expression rate of MDM2, P53 and the negative rate of P16 was 22.9%, 41.7% and 60.4%, respectively. The latter two in high grade (grade III, IV) gliomas had a significantly higher rate than in the low grade (grade II) gliomas. Moreover, the co-expression of MDM2 and P53 protein was confirmed in only 1 of 48 cases. No significant difference was found in the rate of the expression of MDM2 between high grade and low grade gliomas (P > 0.1). PCR-SSCP results showed that mutation of 5-8 exons of p53 gene was detected in 17 out of 48 cases (35.42%). Mutation was detected in 16 of 20 cases of positive p53 expression, and another one was detected in 28 cases of negative expression cases. The correlation between p53 mutation and p53 immunopositivity was observed in 89.6% of the cases. P53 gene mutation and the level of MDM2, P53 and P16 protein were not related to age, gender of the patients, tumor location and size. It is concluded that the mutation of p53 and deletion of p16 might play important roles in the tumorigenesis of gliomas and it was significantly associated with the grade of tumor differentiation. P53 protein accumulation can indirectly reflect p53 mutation. MDM2 amplification and overexpression might be an early event in the growth of human gliomas. PMID- 16696308 TI - Effects of chloroquine on GFAP, PCNA and cyclin D1 in hippocampus and cerebral cortex of rats with seizures induced by pentylenetetrazole. AB - The effects of chloroquine on glial fibrillary acidic protein (GFAP), proliferation cell nuclear antigen (PCNA) and Cyclin D1 in hippocampus and cerebral cortex of rats with seizures induced by pentylenetetrazole (PTZ) were observed in the present study. Forty-eight male adult Sprague-Dawley (SD) rats were randomly divided into control group, chloroquine intervening group, and PTZ group. The behavior and electroencephalogram (EEG) were observed and recorded. GFAP and PCNA were examined with immunohistochemistry. The content of Cyclin D1 in hippocampus and cerebral cortex was inspected with Western blot. The results showed no seizure activity in the control group, severe seizure activity in the PTZ group (IV - V degree), and slight seizure activity (I - IIl degree) in the chloroquine intervening group (P < 0.05). EEG recordings showed no epileptic spikes in the control group, high amplitude with fast frequency in the PTZ group, low-amplitude and slow frequency in the chloroquine intervening group. The expression of GFAP and the positive index of PCNA in the PTZ group were higher than those of control group (P < 0.05 and P < 0.01, respectively). No differences in GFAP expression and PCNA index were observed between chloroquine intervening and control groups (P > 0.05). The content of Cyclin D1 in hippocampus and cerebral cortex was significantly higher in the PTZ group than in control and chloroquine intervening groups (P < 0.05). Therefore, it is considered that chloroquine, by inhibiting the functions and proliferation of glial cells in the hippocampus and cerebral cortex, can alleviate the seizure activities. These results suggest that chloroquine may be an ideal anticonvulsant in preventing and treating epilepsy. PMID- 16696309 TI - Expression of CD151 in human atherosclerotic artery and its implication. AB - To investigate the expression of CD151 in human atherosclerosed artery and explore its clinical implications, Western blot and immunohistochemical techniques were used to determine the protein expression of CD151 in arterial tissues with atherosclerosis taken from 36 patients, including 26 cases who received bypass operation for peripheral artery atherosclerosis and 6 cases who died from coronary heart disease. The expression of CD151 in normal artery tissues from 15 healthy organ donators were also measured to serve as control. The results showed that expression of CD151 protein in atherosclerotic arteries was significantly higher than that in normal artery. In atherosclerotic arteries, CD151 expression was localized in vascular smooth muscle cells (VSMCs) in all tunica media and in partial subintima, while in normal artery, sparse expression was found in tunica media near adventitia. It is concluded that high CD151 protein expression in artery is associated with atherosclerosis and CD151 plays an important role in the atherosclerosis related to VSMC. The expression of CD151 in human atherosclerotic artery depends on the extent of atherosclerotic damage, it's independent of risk factors. PMID- 16696310 TI - Antiapoptotic mechanism of insulin in reoxygenation-induced injury in cultured cardiomyocytes of neonatal rats. AB - To examine the protective effect of insulin on reoxygenation-induced injury and explore the underlying mechanisms, the model of anoxia/reoxygenation (A/R) injury was established by inducing anoxia for 2 h and reoxygenation for 4 h in cultured cardiomyocytes of neonatal rats. The rats were randomized to four groups receiving vehicle, insulin, LY294002, insulin plus LY294002 at the onset of reoxygenation after 2 h of anoxia. At the end of reoxygenation of 4 h, activity of lactate dehydrogenase (LDH) and content of malondialdehyde (MDA) were spectrophotometrically determined, apoptosis of cardiomyocytes were detected by using TUNEL and DNA Ladder, and Western blotting was employed to examine the expression of phosphorylated Akt in all groups. Our results showed that compared with vehicle-treated group, activities of LDH, contents of MDA, apoptosis index (AI) were significantly decreased, and expression of phosphorylated Akt was increased significantly in insulin-treated group. However, changes in LDH, MDA, AI and phosphorylated Akt resulting from insulin were attenuated or abolished by LY294002 (PI3K inhibitor). These data strongly suggest that early administration of insulin at reoxygenation protects cardiomyocytes from reoxygenation-induced apoptosis through PI3K/Akt signaling pathway. PMID- 16696311 TI - Relationship between eukaryotic translation initiation factor 4E and malignant angiogenesis in non-Hodgkin lymphoma. AB - The relationship between angiogenesis and eukaryotic translation initiation factor 4E (EIF4E) expression level in non-Hodgkin lymphoma (NHL) was studied. Mean microvessel density (MVD) and EIF4E were detected in 52 lymph node samples paraffin sections of patients with newly diagnosed NHL by the way of immunohistochemistry. Antisense EIF4E cDNA was cloned into plasmid pcDNA3. 1 (+) and transfected into Raji cells. A series of angiogenesis related factors, including vascular endothelial growth factor (VEGF), matrix metalloproteinases 9 (MMP-9) and tissue inhibitor of metalloproteinases-2 (TIMP-2) proteins were detected by Western blot. The results showed that: (1) The Expression of EIF4E and MVD was higher in aggressive lymphomas than in indolent lymphomas (P < 0.05)and the expression of EIF4E was positively correlated with MVD in lymph node of NHL (r = 0.695, P < 0.01). (2) Antisense EIF4E eukaryocytic expression vector (pcDNA3. 1-EIF4Eas) was constructed successfully. (3) EIF4E, VEGF and MMP-9 were expressed at high levels in Raji cells as compared to normal human peripheral blood monocular cells (NHPMC), and blockage of EIF4E expression brought down the expression of VEGF and MMP-9. However, TIMP-2 was undetectable in Raji cells, although a moderate level of TIMP-2 was detected in NHPMC. It was concluded that the increased EIF4E expression was associated with aggressive property of NHL. PMID- 16696312 TI - Plasma levels of the anti-inflammatory cytokine IL-10 and inflammatory cytokine IL-6 in patients with unstable angina. AB - The plasma levels of inflammatory cytokine interleukin-6 (IL-6) and anti inflammatory cytokine interleukin-10 (IL-10) in the patients with unstable angina or stable angina were determined and compared. In 30 patients with unstable angina and 22 patients with stable angina, plasma levels of IL-10 and IL-6 were detected by ELISA and plasma lipid parameters by lipid research clinical methods respectively. The results showed plasma levels of IL-10 were significantly lower in unstable angina group than in stable angina group (P = 0.005), while those of IL-6 were significantly increased in unstable angina group as compared with those in stable angina group (P = 0.039). There was a significantly negative correlation between IL-10 and IL-6 in patients with unstable angina (r = -0.41, P = 0.003). In the unstable angina group, IL-6 levels were obviously positively correlated with TC (r = 0.314, P = 0.023), but not with TG and HDL. There were no significant correlations between IL-10 and plasma lipid parameters. It was suggested that the decreased IL-10 and increased IL-6 might be associated with the atheromatous plaque stability and progression of coronary heart diseases. IL 10 may play an important role in preventing coronary vascular lesions. PMID- 16696313 TI - Polylactic acid nanoparticles targeted to brain microvascular endothelial cells. AB - In this work, blank polylactic acid (PLA) nanoparticles with unstained surface were prepared by the nano-deposition method. On the basis of the preparation, the effect of surface modification on brain microvascular endothelial cells (BMECs) targeting was examined by in vivo experiments and fluorescence microscopy. The results showed that PLA nanoparticles are less toxic than PACA nanoparticles but their BMECs targeting is similar to PACA nanoparticles. The experiments suggest that drugs can be loaded onto the particles and become more stable through adsorption on the surface of PLA nanoparticles with high surface activity. The surface of PLA nanoparticles was obviously modified and the hydrophilicity was increased as well in the presence of non-ionic surfactants on PLA nanoparticles. As a targeting moiety, polysobate 80 (T-80) can facilitate BMECs targeting of PLA nanoparticles. PMID- 16696314 TI - Effect of PD I administration on dopamine receptors mRNAs expression in the lesioned striatum of PD rat model. AB - To study the effect of PD I administration on dopamine receptors (DR1, DR2) mRNAs expression in the lesioned striatum of the PD rat model and confirm if PD I has the effect of dopamine receptor agonist. The PD rats with unilateral 6 hydroxydopamine lesioned were administrated with PD I, L-dopa methyl/benserazide, L-dopa methyl /benserazide/ PD I, normal saline respectively for 4 weeks and their behavioral changes were observed. Then the rats were sacrificed and RT-PCR technique was used to detect changes of dopamine receptors (DR1, DR2) mRNAs expression in the ipsilateral striatum 1 day after the last treatment. The results showed that treatment with PD I plus L-dopa resulted in a stable contralateral rotation behavior; treatment with L-dopa resulted in a progressively increased contralateral rotation behavior. Rotation behavior induced by anhydromorphine decreased with PD I or PD I plus L-dopa treatment. Treatment With L-dopa or PD I plus L-dopa, up-regulation of DR1 mRNA and down regulation of DR2 mRNA were observed in the ipsilateral striatum which were more obvious than that treated with PD I or vehicle (P < 0.05). It was concluded that long-term treatment with PD I could alleviate the behavior of PD rats. PD I had no apparent effect on the dopamine receptors (DR1, DR2) mRNAs expression in the ipsilateral striatum and the PD I has no agonist effect on dopamine receptors. PMID- 16696315 TI - The influence of the Pro12Ala mutation of PPARgamma2 receptor gene on beta-cells restoration and insulin resistance in type 2 diabetes with hypertension. AB - The aim of this investigation was to determine whether a PPARgamma2 Pro12Ala polymorphism was associated with insulin resistance, beta-cell function and hypertension in Chinese populations. 289 unrelated Chinese subjects first diagnosed Type 2 diabetes (HbAC1 < 6.0) were investigated, including 132 hypertensive diabetic (HTD) subjects, 157 normotensive diabetic (NTD) subjects. Blood pressure and anthropometric measurements were collected from all participants, as well as several venous blood samples during oral glucose tolerance test (OGTT). Biochemical measurements (high-density lipoprotein (HDL) and low-density lipoprotein-cholesterol (LDL), triglycerides) and PPARgamma2 Pro12Ala genotype were also determined. And insulin resistance and beta-cells function was assessed by HOMA-IR and HOMA-beta respectively. The frequency of subjects bearing the Pro12Ala was lower in the hypertension group (3.03%) than in the non-hypertension group (5.7%) (P < 0.05) after adjusted for age, BMI and gender. Hypertensive diabetic Pro12Ala subjects had lower fasting plasma glucose level (P = 0.0127), and better glucose tolerance 60 min after oral glucose (P = 0.0361). Moreover, plasma insulin concentrations at 60 min was lower than those without A variant (P = 0.0275), and both hypertensive Ala/Pro in HOMA-beta (P = 0.0455) and AUC for insulin (P = 0.0473) were higher, and HOMA-IR was lower (P = 0.0375) as compared with hypertensive Pro/Pro subjects. No association was observed between Pro12Ala genotype and BMI, total cholesterol, HDL- cholesterol or triglycerides in either group. Our findings suggested that the Ala 12 allele of the PPARgamma2 gene may improve insulin resistance and ameliorate beta-cell function reserves in T2DM with hypertension, and protect patients from hypertension in T2DM. As an important thrifty gene, environment factors may exerts an effect of PPAR gamma2 on glucose homeostasis and insulin resistance. PMID- 16696316 TI - Expression of serum and glucocorticoid-inducible kinase1 in diabetic rats and its modulation by fluvastatin. AB - The expression of serum and glucocorticoid-induced protein kinase in the renal cortex of diabetic rats was examined, and the function of signal transduction mediated by SGK1 in diabetic nephropathy and its modulatiqn by fluvastatin were also investigated. 24 male Wistar rats were randomly divided into normal control group (n = 8), diabetic nephropathy group (n = 8) and fluvastatin-treated diabetic nephropathy group (15 mg/kg/d, n = 8). The metabolic parameters were measured at the 8th week. The expression of transforming growth factor beta1 (TGF beta1) and fibronectin (FN) was immunohistochemically examined. The expression of SGK1 was detected by RT-PCR and Western blot, and CTGF mRNA was assessed by RT PCR. As compared to DN, blood glucose, 24-h urinary protein, Cer and kidney weight index were all decreased and the weight was increased obviously in group F. At the same time, mesangial cells and extracellular matrix proliferation were relieved significantly. The levels of cortex SGK1 mRNA and protein were up regulated, and both TGF-beta1 and FN were down-regulated by fluvastatin. The mRNA of SGK1 was positively correlated with the CTGF, TGF-beta1 and FN. SGK1 expression is markedly up-regulated in the renal cortex of DN group and plays an important role in the development and progress of diabetic nephropathy by means of signal transduction. Fluvastatin suppressed the increased SGK1mRNA expression in renal cortex and postponed the development of diabetic nephropathy. PMID- 16696317 TI - RNA interference targeting leptin gene effect on hepatic stellate cells. AB - To construct the specific siRNA expression vectors and investigate their effect on leptin and collagen I in HSC, which provide a new approach to the prevent and treat hepatic fibrosis. The five siRNAs against leptin gene were transcript synthesized intracellularly by expression templates of plasmid vector psiRNA hH1neo. The recombinant leptin siRNA plasmid vectors could express in eukaryocyte, and then to evaluate them by using enzyme cutting and sequencing. The recombinant plasmids were transfected into HSCs using Lipofectamine methods respectively. The cells were selected after growing in DMEM containing 300 microg/mL G418 for about 4 weeks. Gene expression of leptin and collagen I were showed by Western blot analysis and reverse transcription polymerase chain reaction (RT-PCR). Identification by enzyme cutting and sequencing showed that the leptin siRNA expression vectors were constructed successfully, and leptin siRNA could inhibit the leptin and collagen I gene expression effectively. It was concluded that RNA interference-mediated silencing of leptin gene diminished leptin and collagen I gene expression in HSCs. Furthermore, attenuated the extracellular matrix over-deposition at the same time. Leptin gene is ideal targets of gene therapy for liver fibrosis. PMID- 16696318 TI - Effect of antisense MBD1 gene eukaryotic expression plasmid on expression of MBD1 gene in human biliary tract carcinoma cells. AB - Hypermethylation of the promoter region is one of the major mechanism of tumor suppressor gene inactivation. In order to provide a research tool for the study on the function of MBD1 gene in DNA methylation and tumorigenesis, antisense MBD1 gene eukaryotic expression plasmid was constructed and transfected into human biliary tract carcinoma cell line QBC-939 to observe its effect on the expression of MBD1 mRNA and protein by using RT-PCR and FCM respectively. Following the transfection, the mRNA level of MBD1 gene decreased from 0. 912 +/- 0.022 to 0.215 +/- 0. 017, and the protein level of MBD1 gene also decreased from (80.19 +/- 5.05) % to (35.11 +/- 4.05) %. There were very significant differences in the expression both at the transcription and post-transcription levels of MBD1 gene between non-tranfection group and the antisense MBD1 gene eukaryotic expression plasmid transfection group (P < 0.01). It was suggested that transfection with the antisense MBD1 gene eukaryotic expression plasmid can significantly reduce the expression level of MBD1 gene in QBC-939, and this study may provide a valid tool for the investigation of the function of MBD1 gene and its role in biliary tract carcinoma. PMID- 16696319 TI - Reversal of HCC drug resistance by using hammerhead ribozymes against multidrug resistance 1 gene. AB - To reverse multidrug resistance(MDR) of HepG2 by anti-MDR1 hammerhead ribozyme, an anti-MDR1 hammerhead ribozyme was developed and delivered to P-gp overproducing human hepatocarcinoma cell line HepG2 by a retroviral vector containing RNA polymerase III promoter. The expression of mdr1/Pgp and Rz was detected in HepG2, HepG2 multidrug-resistant cell line and HepG2 Rz-transfected cells by semi-quantitative RT-PCR and Western blot methods. Moreover, MTT assay was employed to detect the sensitivity of these ribozyme-transfected cells, and Rhodamine123 (Rh123) was used to test the function of Pgp. The Rz- transfected HepG2 cells became doxorubicin-sensitive, which was concomitant with the decreased MDR1 expression. The study showed that the retrovirus vector encoding the anti-MDR1 ribozyme may be applicable to the treatment of MDR cells. PMID- 16696320 TI - IL-5 up-regulates the expression of TGF-beta1 in human blood eosinophils in vitro. AB - To investigate the effects of IL-5 on the expression of TGF-beta1 in eosinophils in vitro, eosinophils were incubated in the presence of the same concentrations of 1L-4, IL-5 and IFNgamma, different concentrations of IL-5 in vitro and changes of eosinophil viability were assessed by trypan blue exclusion. Non-cytokine was employed as a negative control. 16 h after the cultivation, supernatants and cells were assayed by using TGF-beta1 specific ELISA and RT-PCR. The mRNA expression and protein expresssion of TGF-beta1 in eosinophils stimulated with different cytokines was observed. The expression of TGF-beta1 protein in eosinophils was increased significantly by IL-4 (433.67 +/- 9.86 vs 228.9 +/- 2.87) and IL-5 (403.72 +/- 7.60 vs 228.9 +/- 2.87, P < 0.05), while decreased by IFNgamma (178.47 +/- 2.60 vs 228.9 +/- 2.87). At the same time, the results demonstrated that the basal level of TGF expression was enhanced by IL-5 in all samples (P < 0.05). The expression of TGF-beta1 mRNA was 1.42, 1.70, 1.76-folds higher than that of the non-stimulated controls. It is concluded that IL-5 can up regulate the expression of TGF-beta1 in eosinophils in vitro, which might have effect in eosinophil-associated chronic rejection. PMID- 16696321 TI - Preparation of curcumin prodrugs and their in vitro anti-tumor activities. AB - The curcumin prodrugs, which could be selectively activated in tumor cells, were prepared to establish a basis for the targeted chemotherapy for cancer. On the basis of the molecular structure of curcumin, the N-maleoyl-L-valine-curcumin (NVC), N-maleoyl- glycine-curcumin (NGC) were chemically synthesized and identified by IR and NMR spectroscopy. After treatment with these two prodrugs for 6 - 24 h, the rates of growth inhibition on human bladder cancer EJ cells and renal tubular epithelial (HKC) cells were detected by MTT colorimetry. Our results showed that after the treatment with 20 micromol/L - 40 micromol/L NVC and NGC for 6 - 24 h, the growth inhibitory effects on EJ cells were 6.71% - 65.13% (P < 0.05), 10.96% - 73.01% (P < 0.05), respectively, in both dose- and time-dependent manners. When compared with the curcumin of same concentrations, the growth inhibitory effects of these two prodrugs on HKC cells were significantly decreased (P < 0.01). It is concluded that activation of curcumin prodrugs via hydrolysis functions of cellular esterase could inhibit the growth activities of tumor cells, and reduce the side effects on normal diploid cells. This provided a novel strategy for further exploration of tumor-targeted chemotherapeutic drugs. PMID- 16696322 TI - Immunohistochemical analysis of Omi/HtrA2 expression in prostate cancer and benign prostatic hyperplasia. AB - To study the expression and significance of the serine protease Omi/HtrA2 in prostate cancer and benign prostatic hyperplasia. The expression of Omi/HtrA2 was assayed by means of immunohistochemical technique in 41 prostate cancer (Cap), 20 benign prostatic hyperplasia (BPH) and 10 normal prostate (NP) specimens. Omi/HtrA2 expression was positive in 30 (73.17%) prostate cancer specimens, and the positive rate of Omi/HtrA2 was lower in well differentiated than in poorly and moderately differentiated groups (P < 0.05). By contrast, the cells in normal prostate and benign prostatic hyperplasia groups showed no or weak expression of Omi/HtrA2. Prostate cancer cells in vivo may need Omi/HtrA2 expression for apoptosis, and that Omi/HtrA2 expression might be involved in prostate cancer development. PMID- 16696323 TI - Wild-type Smad3 gene enhances the osteoblastic differentiation of rat bone marrow derived mesenchymal stem cells in vitro. AB - This study examined the effect of wild-type Smad3 gene on the osteoblastic differentiation of rat bone marrow-derived mesenchymal stem cells in vitro. Bone marrow-derived mesenchymal stem cells (MSCs) were stably transfected with the complexes of pcDNA3. 0-Myc-Smad3 or pcDNA3. 0-Myc-Smad3deltaC and Lipofectamine reagent. Immunofluorescence staining was performed to evaluate the c-Myc signal in MSCs. The cell proliferation was detected by MTT method. To clarify the osteoblastic characteristics in stably transfected MSCs, alkaline phosphatase (ALP) mRNA and core binding factor alpha1 (Cbfa1) mRNA were investigated by RT PCR, and ALP activity and mineralization were examined by p-nitrophenolphosphate method and alizarin red staining respectively. PD98059, a specific inhibitor of the ERK signaling pathway, was used to determine the role of ERK in Smad3-MSCs osteoblastic differentiation. c-Myc signal was detected in Smad3-MSCs and Smad3 deltaC-MSCs. The proliferation of Smad3-MSCs was slower than that of Smad3deltaC MSCs or V-MSCs. The relative levels of ALP mRNA and Cbfal mRNA in Smad3-MSCs, as well as ALP activity and mineralization, were markedly higher than those in Smad3deltaC-MSCs or V-MSCs. Although ALP activity and mineralization were slightly lower in Smad3-MSCs treated with PD98059 than in those without PD98059 treatment, no significant difference was found between them (P > 0.05). It is concluded that the wild-type Smad3 gene, which is a crucial component promoting bone formation, can inhibit the proliferation of MSCs and enhance the osteoblastic differentiation of uncommitted MSCs and the maturation of committed MSCs independent of the ERK signaling pathway. PMID- 16696324 TI - Effect of exogenous p16ink4a and hRb1 genes on cell cycle regulation of osteosarcoma cell. AB - To study the effect on regulation of cell cycle of osteosarcoma cell line MG63 tranceduced with exogenous p16ink4a and hRb1 genes, pIRES-p16ink4a-hRb1, pIRES p16ink4a and pIRES-hRb1 plasmids were constructed by gene recombination technology. The recombinant plasmid was transferred into osteosarcoma cell line MG63 by metafectene, and the resistant clones were selected by G418 selective medium. mRNA and protein expression of osteosarcoma cell line were assayed by RT PCR and Western-Blot respectively. Cell cycle and apoptosis were analyzed by subG1 flow cytometric. Cell proliferation was tested by MTT. In the genome of these transfected target cells, the expression of p16ink4a and hRb1 mRNA and protein were detected respectively in vitro. It was demonstrated with subG1 flow cytometric analysis and MTT method that p16ink4a and hRb1 genes cooperation more significantly inhibited cell growth and induced a more marked G1 arrest and apoptosis than p16ink4a/hRb1 alone (P < 0.01). Coexpression of exogenous p16ink4a with hRb1 broke the regulatory feedback loop of p16ink4a-cyclinD1 /CDK-hRb1 and played a more significant role in inhibiting cell growth as well as inducing cell apoptosis than p16ink4a or hRb1 did alone in vitro. PMID- 16696325 TI - The relationship of the expression of estrogen receptor in cartilage cell and osteoarthritis induced by bilateral ovariectomy in guinea pig. AB - To investigate the estrogen receptor (ER) expression in cartilage cell in the development of osteoarthritis induced by bilateral ovariectomy in guinea pig and to find their relationship. 30 two-month-old female guinea pigs were randomly divided into two groups (n = 15 each): sham operation (control) group and ovariectomized group (OVX); Scanning electorne microscope (SEM) and transmission electron microscope (TEM) were obtained to analysis the cartilage degeneration of the hind limb knee joint after 6 and 12 weeks of ovariectomy. Dextran-Coated Charcoal (DCC) was taken to quantitively detect the expression of ER. The serum levels of estrogen and gestone were detected by immune contest assay. The results showed that ER do exist in the cartilages of the guinea pigs, with higher expression in the control group than in OVX group at the same time point (P < 0.05). It was increased also at 12 th week after operation than that of preoperation. The blood serum levels of estrogen and gestone showed a similar tendency to the expression of ER. Joint cartilage degeneration detected by SEM and TEM could be found at 6 th week, but severe degenerative lesions at 12 th week in the OVX group compared with the control group (P < 0.01). The data suggested that bilateral ovariectomy in guinea pig lead to severe osteoarthritis which mighgt be related to the lower serum level of estrogen and the downregulation of the expression of ER in the cartilage also. PMID- 16696326 TI - The construction and investigation of PLGA artificial bone by biomimetic mineralization. AB - To modify the surface property of poly lactide-co-glycolide (PLGA) by biomimetic mineralization to construct a new kind of artificial bone. PLGA films and 3 diamensional (3-D) porous scaffolds hydrolyzed in alkaline solution were minerilized in SBF for 14 days. The morphology and composition of the mineral grown on PLGA were analyzed with SEM, FTIR and XRD. The porosity of the scaffolds was detected by using the liquid displacement method. The compressive strength of the scaffolds was detected by using a Shimadzu universal mechanic tester. An obvious mineral coating was detected on the surface of films and scaffolds. The main component of the mineral was carbonated hydroxyapatite (HA) similar to the major mineral component of bone tissues. The porosity of the un-mineralized and mineralized porous scaffolds was (84.86 +/- 8.52) % and (79.70 +/- 7.70) % respectively. The compressive strength was 0.784 +/- 0.156 N/mm2 in un mineralized 3-D porous PLGA and 0.858 +/- 0.145 N/mm2 in mineralized 3-D porous PLGA. There were no significant differences between the mineralized and un mineralized scaffolds (P > 0.05) in porosity and biomechanics. Biomimetic mineralization is a suitable method to construct artificial bone. PMID- 16696327 TI - Effects of Icariin on expression of OPN mRNA and type I collagen in rat osteoblasts in vitro. AB - To study the effects of Icariin on expression of osteopontin (OPN) mRNA and type I collagen in rat osteoblasts in vitro and to explore its possible mechanisms in preventing osteoporosis. OB was isolated from calvaria of new-born new-born fetal Sprague-Dawley (SD) rats by means of modified sequential collagenase digestion and incubated in MEM medium and the cell morphology was observed under inverted phase contrast microscope, OB was identified by alkaline phosphatase (ALP) staining. Different concentration (0.1 microg/mL, 1.0 microg/ml, 10 microg/mL) of Icariin was added to the OB and incubated. The effect of Icariin on the proliferation and osteogenesis of OB was monitored by MTT analysis. The expression of type I collagen was estimated with immunohistochemistry techniques. The expression levels of mRNA of OPN in the cells in every group were examined by reverse-transcriptase ploymerase chain reaction (RT-PCR). The expression of OPN mRNA and type I collagen was strengthened gradually with the increase of Icariin concentration and peaked with 10 microg/mL Icariin on the 5th day. Icariin could significantly promote the expression of OPN mRNA and type I collagen in rat osteoblasts in vitro. The levels of expression of OPN mRNA and type I collagen were changed with different concentration of Icariin. Icariin could effectively prevent and treat osteoporosis and promote the bone formation. PMID- 16696328 TI - Expression of heat shock protein 70 and 27 in non-small cell lung cancer and its clinical significance. AB - The heat shock proteins (HSPs) 70 and HSP 27 expression in patients with non small cell lung cancer (NSCLC) was studied and the relationship between HSP 70 and HSP 27 with the clinicopathological features of NSCLC was investigated. The expression of HSP 70 and HSP 27 was detected in tumor tissues from 60 patients with NSCLC by S-P immunohistochemistry. The findings were analyzed in combination with the histological types, histopathological differentiation, lymph node metastasis, patients' clinical stages, smoking history and gender. The results showed that of the 60 NSCLC tissue specimens studied, the immunoreactivity of HSP 70 and HSP 27 was detected in 47 (78.3%) and 43 (71.7%) specimens, respectively. A positive correlation was found between the overexpression of HSP 70 and HSP 27. The histopathological differentiation, lymph node metastasis, clinical stages and smoking history were correlated to the expression of HSP 70, but not to the expression of HSP 27. No statistical significance was observed in histological types and gender with respect to both HSP 70 and HSP 27 expression. It is suggested that the HSP 70 expression is a powerful and significant prognostic indicator and is related to histopathological differentiation, lymph node metastasis, patients' clinical stages, smoking history, whereas HSP 27 expression is not. PMID- 16696329 TI - Effects of intrathecally administerd NaV1. 8 antisense oligonucleotide on the expression of sodium channel mRNA in dorsal root ganglion. AB - Neuropathic pain has been hypothesized to be the result of aberrant expression and function of sodium channels at the site of injury. To investigate the effects of NaV1. 8 antisense oligonucleotide on the expression of sodium channel mRNA in dorsal root ganglion (DRG) neurons in chronic neuropathic pain. 24 Sprague-Dawley rats weighing 200-260 g were anesthetized with the intraperitoneal injection of 300 mg x kg(-1) choral hydrate. The CCI model was made by loose ligation of sciatic nerve trunk by 4-0 chromic gut. The mechanical and thermal pain threshold were measured before operation and 1, 3, 5, 7, 9, 11, 13 days after operation. A PE-10 catheter was implanted in subarachnoid space at lumbar region. On the 7th postoperative day the animals were randomly divided into 4 groups. The drugs were injected intrathecally twice a day for 5 consecutive days in group 2-4. The animals were decapitated 14 days after the surgery. The L4-L6 DRG of the operated side was removed and crushed, and total RNA was extracted with Trizol reagent. The contralateral side was used as control. The change of NaV1. 8 sodium channel transcripts was determined by RT-PCR. Pain threshold was significantly lowered after CCI as compared with that in control group and was elevated 3 days after antisense oligonucleotide injection. Sensory neuron specific TTX-R sodium channel NaV1. 8 transcript was down-regulated after antisense oligonucleotide injection at the dosage of 45 microg as compared with that in CCI group (P < 0.01), and it was even greater at the dosage of 90 microg. The intrathecally injected NaV1. 8 antisense oligonucleotide can reduce the mechanical allodynia and thermal hyperalgesia partially by downregulating the SNS transcript expression. PMID- 16696330 TI - Effect of propofol on glutamate and gamma-aminobutyric acid release from rat hippocampal synaptosomes. AB - To investigate the effect of propofol on the release of glutamate and gamma aminobutyric acid (GABA) from rat hippocampal synatosomes, synaptosomes was made from hippocampus and incubated with artificial cerebrospinal fluid (aCSF). With the experiment of Ca(2+)-dependent release of glutamate and GABA, dihydrokainic acid (DHK) and nipectic acid were added into aCSF. For the observation of Ca(2+) independent release of glutamate and GABA, no DHK, nipectic acid and Ca2+ were added from aCSF. The release of glutamate and GABA were evoked by 20 micromol/L veratridine or 30 mmol/L KCI. The concentration of glutamate and GABA in aCSF was measured by using high-performance liquid chromatography (HPLC). 30, 100 and 300 micromol/L propofol significantly inhibited veratridine-evoked Ca(2+)-dependent release of glutamate and GABA (P < 0.01 or P < 0. 05). However, propofol showed no effect on elevated KCl-evoked Ca(2+)-dependent release of glutamate and GABA (P > 0.05). Veratridine or elevated KCI evoked Ca(2+)-independent release of glutamate and GABA was not affected significantly by propofol (P > 0.05). Propofol could inhibit Ca(2+)-dependent release of glutamate and GABA. However, it has no effect on the Ca(2+)-independent release of glutamate and GABA. PMID- 16696331 TI - Assessment of left ventricular longitudinal regional myocardial systolic function by strain imaging echocardiography in patients with hypertrophic cardiomyopathy. AB - To assess the left ventricular longitudinal regional myocardial systolic function by strain imaging (SI) echocardiography and to study the relationship between regional myocardial systolic function and left ventricular structure in patients with hypertrophic cardiomyopathy (HCM). SI echocardiography were performed in 18 patients with HCM and 17 healthy subjects. For each wall, regional myocardial systolic strain was analyzed at the basal, mid, and apical level respectively. And the peak systolic strain was measured. Our results showed that the patients with HCM had reduced peak systolic strain at almost each segment of different walls when compared with healthy subjects. There was significant correlation between the mid-septum peak systolic strain and the thickness of IVS, so was the correlation between the mid-septum peak systolic strain and the IVS to LVPW thickness ratio. This study demonstrated that the left ventricular longitudinal regional myocardial systolic function was abnormal in HCM, and this kind of abnormalities existed extensively in hypertrophic and non-hypertrophic cardiac segments. The degrees of left ventricle hypertrophy and asymmetry are related to the myocardial regional systolic function in HCM. PMID- 16696332 TI - [Experimental study on the effect of transarterial chemoembolization with Bletilla striata in liver neoplasm]. PMID- 16696333 TI - Role of beta-adrenergic signal transduction pathway on myocardial ischemic preconditioning of rats. AB - To study the changes in every part of the beta-adrenergic signal transduction pathway and their effects on ischemic preconditioning of rat myocardium in vivo. SD rats were divided into three groups: IP group, I/R group and CON group. The IP group was further divided into PC1-, 2-, 3-, and PC1+, 2+, 3+ groups according to preconditioning procedure. The rats received surgical procedure and underwent left coronary artery occlusion and reperfusion. We analyzed the infarct size by TTC staining, measured serum myocardial enzymes, studied the beta-AR Bmax and Kd by radioligand binding assay of receptors, checked the activity of AC and PKA by the method of biochemistry and examined the content of cAMP by radioimmunoassay. The infarct area was much smaller in the IP group than in the I/R group (P < 0.001), while the enzymes were significantly higher in I/R (P < 0.001). The Bmax of beta-AR in IP was much higher than that in I/R (P < 0. 001), but no difference in Kd could be seen between IP and I/R groups. In IP, the activity of AC and PKA and the content of cAMP were higher than those in I/R (P < 0.05, 0.002 and 0.001, respectively). In the procedure of preconditioning, the content of cAMP and the activity of PKA showed the characteristic of cyclic fluctuation. Ischemic preconditioning can protect the heart from necrosis and reduce endo-enzyme leakage. The system of beta-adrenergic signal transduction pathway probably takes part in the protection effect of the IP, which might be elicited by the PKA. PMID- 16696334 TI - A comparative study of 99mTc-YIGSR and 99mTc-MIBI uptake in tumor cells. AB - To investigate a new kind of tumor tracer 99mTc-YIGSR developed from a five amino structure (YIGSR) of the Laminin -chain, which can bind to the laminin receptors of tumor specifically, and radiolabeled with MAG3. (1) Preparation of the 99mTc YIGSR probe: with S-Acetly-NH3-MAG3 as the chelator and with proper reductants YIGSR was labeled with 99mTc; (2) Cell culture and viability measurement: EAC was maintained in RPMI 1640 supplemented with calf serum; the trypan blue exclusion was applied to calculate the cell viability; (3) Study of the cell dynamic: The EAC's uptake of 99mTc-YIGSR and 99mTc-MIBI was observed at 37 degrees C and 22 degrees C, respectively. (1) The labeling efficiencies of 99mTc-YIGSR and 99mTc MIBI were (62 +/- 3)% and (96 +/- 2)%, respectively; (2) The cell viability was declined with time of incubation; (3) At 37 degrees C, the EAC'S uptake of 99mTc YIGSR and 99mTc-MIBI reached the peak of (43.16 +/- 2.4) % and (24.4 +/- 1.8) % at 60 min, respectively; and at 22 degrees C, the highest uptake was (26.5 +/- 2.1) % and (9.47 +/- 1.9) % at 60 min, respectively. The in vitro study suggests that 99mTc-YIGSR is superior to 99mTc-MIBI in cell uptake and has potential value in tumor imaging. PMID- 16696335 TI - Construction of antisense MT1-MMP vector and its inhibitory effects on invasion of human ovarian cancer cells. AB - Membrane-type 1 matrix metalloproteinase (MT1-MMP/MMP-14) plays crucial roles in tumor cell growth, invasion, and angiogenesis. To clarify whether the endogenously expressed MT1-MMP in metastatic human ovarian carcinoma cell lines SKOV3 plays a critical role in tumor cell invasiveness, antisense MT1-MMP cloned in eukaryotic expression vector pMMP14as was transferred into SKOV3 cells. 48h after transfection, decreased expression of endogenous MT1-MMP protein was detected in pMMP14as-transfected SKOV3 cells and the activation of pro-MMP2 was inhibited markedly. The mean percentage of invasive cells was (62.50 +/- 5.30) % in pMMP14as-transfected cells, which was obviously less than that (97.20 +/- 6.90) % in the control. Thus, antisense MT1-MMP effectively inhibited the endogenous MT1-MMP expression and the invasiveness in SKOV3 cells, suggesting that MT1-MMP may be a therapeutic target molecule for human invasive ovarian cancers. PMID- 16696336 TI - The effects of magnesium sulfate on fetal rats of FGR and the expression of caspase-3 in the placenta of maternal rat. AB - To investigate the effect of magnesium sulfate on the fetal rats of FGR and the expression of caspase-3 in the placenta of maternal rat; To explore the mechanism of using magnesium sulfate to cure the FGR. Establish model of FGR by a way of passive smoking: giving the maternal rats different agent of magnesium sulfate by subcutaneous injection: low agent group (300 mg/kg), high agent group (600 mg/kg). Concentration of magnesium sulfate was monitored. The expression of caspase-3 was measured by RT-PCR and immunohistochemistry technology. Both of the concentrations of magnesium sulfate in high and low agents group are higher than the FGR group (P < 0.01); the weight of the placenta and fetal rat in high agent group are higher than the FGR group (P < 0.05 and P < 0.01); the expression of mRNA and protein of caspase-3 in the two agent group is higher than the FGR group (P < 0.05 respectively); concentration of magnesium sulfate in the maternal rat blood correlate to the weight of fetal rat (r = 0.899, P = 0.038) and the expression of caspase-3 in the placenta of maternal rat (r = -0.747, P = 0.033; r = - 0.915, P = 0.001). The research suggests that the weight of fetal rat could be increased by treatment of magnesium sulfate. Because it would imfrmove the placental function by depressing the expression of caspase-3. PMID- 16696337 TI - A study on the traditional Chinese medicine Jinyebaidu for prevention and treatment of intrauterine infection with guinea pigs cytomegalovirus. AB - The purpose is to study the prophylactic and therapeutic effect of the traditional Chinese Medicine (TCM)-Jinyebaidu (JYBD) to guinea pig cytomegalovirus (GPCMV) intrauterine infection. The virus-free female and male guinea pigs were screened with nest-polymerase chain reaction (N-PCR). After inbred, pregnant guinea pigs were selected and divided into 3 groups randomly: 5 guniea pigs of the blank control group were not given either GPCMV or JYBD. 31 guniea pigs of the positive control group were inoculated 1 mL (10(7) TCID50) suspension of GPCMV intraperitoneal. 10 guniea pigs of the experimental group were inoculated GPCMV firstly and then perfused stomach with JYBD for 14 days (Dosage in accordance with the modulus of the weight ratio of human to guniea pig). The effects of JYBD on the intrauterine infection of GPCMV were observed. The results showed that JYBD could decrease the maternal infection rate from 100% (31/31) to 50% (5/10) (P < 0.001), the intrauterine infection rate from 100% (72/72) to 75% (21/28) (P < 0.001), and the rate of abnormal outcome of pregnancy from 64.4% (29/45) to 25.0% (7/28) (P < 0.001), the infective symptoms being relieved. It can be concluded that traditional Chinese medicine- JYBD can prevent and treat (GPCMV intrauterine infection, and can be expected a prophylactic drug for HCMV intrauterine infection. PMID- 16696338 TI - Serum IL-10 level in allergic rhinitis patients and its effect on serum total IgE. AB - To investigate the level of serum IL-10 in allergic rhinitis patients and the correlation between IL-10 and serum total IgE (TIgE). 50 allergic rhinitis patients and 30 normal subjects were involved in the study. The levels of serum IL-10 and TIgE were measured by enzyme-linked immunosorbent assay (ELISA), and the correlation between serum IL-10 and TIgE was analyzed. In the allergic rhinitis group, the levels of serum IL-10 and TIgE were 8.34 +/- 2.48 pg/mL and 142.6 +/- 28.2 KU(A)/L. In the normal control group, they were 12.86 +/- 2.88 pg/mL and 47.2 +/- 12.2 KU(A). There were significant differences between the two groups (P < 0.01); and the level of serum TIgE in the patients was negatively correlated with that of IL-10 (r = -0.46, P = 0.02). The level of serum IL-10 was significantly decreased in allergic rhinitis patients, which was beneficial to the synthesization of IgE. IL-10 plays an important role in the episode of allergic rhinitis. PMID- 16696339 TI - Study on remodeling of astrocytes in facial neuclus after peripheral injury. AB - To observe the glial reactions surrounding facial motor neurons following facial nerve anastomosis. At 1, 7, 21 and 60 d following facial nerve anastomosis, the recovery process of facial movement was observed, the glial fibrillary acidic protein (GFAP) immunoreactivity was analyzed by a combined method of fluorescent retrograde tracing and immunofluorescent histochemical staining, and the ultrastructure of astrocytes were observed under a transmission electron microscope (TEM), respectively. Postoperatively the function of facial muscles could not return to normal, often accompanied with hyperkinetic syndromes such as synkinesis at the late stage. Motor neurons in every facial subnucleus could be retrogradely labeled by fluoro-gold (FG), and displayed an evident somatotopic organization. Normally there was a considerable number of GFAP-positive cells in nonnucleus regions but few inside the facial nucleus region. Postoperatively the GFAP immunoreactivity in the anastomotic side increased significantly, but gradually decreased at the late stage. The ultrastructure of astrocytes in our experiment showed that the sheet-like process of astrocytes invested and protected the injured facial motor neurons. The present study shows that reactive astrocytes undergo some characteristic changes during the process of facial nerve injury and regeneration. The plastic change at the late stage may be involved in the mechanism of synkinesis. PMID- 16696340 TI - Inhibitory effect of tissue transglutaminase (tTG) antisense oligodeoxynucleotides on tTG expression in cultured bovine trabecular meshwork cells. AB - To study the effect of tTG fully phosphorothioated antisense oligodeoxynucleotides (tTG-ASDON) on tTG expression in cultured bovine trabecular meshwork cells (BTMCs) in vitro and explore a new treatment alternative for primary open angle glaucoma (POAG), the ASDON1 and ASDON2 complementary to the protein codogram region of tTG were designed, synthesized and phosphorothioated according to the secondary structure of tTG. The ASDON1 and ASDON2 were embedded in Lipofectamine and transfected into BTMCs. The untreated group served as negative controls. The expression of tTG in the mRNA and protein level were measured by semi-quantitative RT-PCR and immunohistochemical technique Supervision method respectively. Our results showed that both the mRNA and the protein of tTG with tTG-ASDON and tTG-ASDON2 were significantly decreased as compared with that of the controls (P < 0.05). On the other hand, no significant difference was found between the ASDON1 group and the ASDON2 group. It is concluded that the expression of tTG mRNA and protein in cultured BTMC are down regulated by tTG- ASDON. As a result, tTG-ASDON may be used for the treatment of POAG through the inhibitory effect on the expression of tTG. PMID- 16696341 TI - Effect of Homoharringtonine on corneal haze after excimer laser photorefractive keratectomy in rabbits. AB - To evaluate the inhibiting effect of Homoharringtonine (HHT) on the corneal haze after excimer laser photorefractive keratectomy (PRK) in rabbits. 18 healthy rabbits which underwent PRK were randomly divided into three groups (A, B and C). The refractive degree of ablation was - 10.0DS in each group. Group A was locally treated with a piece of filter paper soaked with 1 mg/mL HHT for 5 min, and then the entire cornea was repeatedly irrigated with balance solution; Group B was dropped with 0.1 mg/mL HHT after PRK for 3 months; Group C was the control group. Corneal haze, histopathology, response, ect. were investigated. The corneal haze was significantly less in group A, while the difference between group B and group C was insignificant. Keratocytes and fibrocytes in corneal stroma were more active up to 3 months in group B and group C. Intraoperative use of topical HHT can reduce corneal haze after PRK in rabbits. PMID- 16696342 TI - Regulatory effect of dexamethasone on aquaporin-1 expression in cultured bovine trabecular meshwork cells. AB - To evaluate the effect of dexamethasone on the expression of aquaporin-1 (AQP-1) in cultured bovine trabecular meshwork cells, bovine trabecular meshwork cells were cultured in vitro and reproduced to the third and the fourth generation, then treated with dexamethasone at the concentrations of 5, 25, 50, 250 microg/L respectively for 7 days. Immunohistochemical technique-supervision method was employed to measure, and image analysis system to analyze the expression of AQP-1 in normal cultured bovine trabecular meshwork cells and those treated with dexamethasone. In normal bovine trabecular meshwork cells, the grayscale of AQP-1 positive staining was 167.94 +/- 1.18, while it was 168.92 +/- 0.91, 176.72 +/- 1.80, 180.64 +/- 1.31, 185.64 +/- 1.58 in cells treated with 5, 25, 50, 250 microg/L concentrations of dexamethasone. When the concentration of dexamethasone was higher than 25 microg/L, the expression of AQP-1 was significantly inhibited (P < 0.05). The regulation of AQP-1 expression by dexamethasone in cultured bovine trabecular meshwork cells in vitro may be one of causes that retard the aqueous outflow in glucocorticoid- induced glaucoma. PMID- 16696343 TI - Effect of cyclosporin A eye drop on keratoconjunctivitis sicca and its mechanism. AB - In this study, the effect of cyclosporin A (CsA) eye drop on keratoconjunctivitis Sicca (KCS) and its mechanism were studied. The KCS models were established by injecting Pertussis vaccine, complete Freund's adjuvant (CFA) and antigen of conjunctiva from isotype mice. Then the KCS models were treated with cyclosporin A eye drop. Changes in breaking-up time (BUT), lacrimal secretion in 30 min and diversion in 24 h were measured. The percentage of beaker cells, the lymphocytic infiltration in conjunctiva were observed. The expression levels of Aquaporin-3 (AQP3) in conjunctiva epithelial cells, beaker cells and accessory lacrimal gland were immunohistochemically detected. The results showed that there were significant differences in BUT, the percentage of beaker cells, lacrimal secretion in 30 min, the lymphocytic infiltration and the expression of AQP3 between the experimental group and an control group. It was concluded that CsA eye drop exerts marked therapeutic effect on KCS by inhibiting T lymph cells, increasing the goblet cells and AQP3 expression in conjunctiva. PMID- 16696344 TI - Expression of telomerase subunits in gastric cancer. AB - To detect the expression of telomerase subunits (human telomerase reverse transcriptase, human telomerase associated protein 1 and human telomerase RNA) in gastric cancer and to examine the role that different telomerase subunits play in the gastric carcinogenesis, reverse transcription-polymerase chain reaction (RT PCR) was used to detect telomerase subunits messenger RNA in 24 samples of gastric cancer and corresponding non-cancerous tissue. The results showed that the positive rate of hTERT mRNA from gastric cancer and corresponding non cancerous tissues was 100% and 25%, respectively. The former was significantly higher than the latter (chi2 = 26.4, P < 0.01). The positive rate of hTEP1 mRNA from gastric cancer and corresponding non-cancerous tissues was 100% and 91.7%, respectively and no significant difference was found between them (chi2 = 2.1, P > 0.05). The positive rates of hTR for gastric cancer and corresponding non cancerous tissues were both 100% and no significant difference existed between them. It is concluded that in contrast to hTEP1 and hTR, the up-regulation of hTERT mRNA expression may play a more important role in the development of gastric cancer. PMID- 16696345 TI - Susceptibility to vaginal candidiasis under different conditions in mice. AB - In order to study the susceptibility of murine vaginal mucosa to Candida albicans under different conditions, vaginal lavage fluid and vaginal tissue of mice were observed and compared between murine models with normal immune system (estrogen treated mice) and immunosuppressed murine model, and between primary infection model of vaginal candidiasis and secondary infection one. The average level of colony forming unit (CFU) from the immuosuppressed group was higher than that from estrogen-treated group at each time point and the peak time was delayed. The differences between the two groups were statistically significant (P < 0.05) from the fourth day after inoculation. A significant difference existed in the average level of CFU between the control group and the estrogen-treated group (P < 0.05), and between the control group and the immuosuppressed group (P < 0.01). It was concluded that the vaginal mucosa from the immunosuppressed mice is more susceptible to Candida albicans and no difference is found in susceptibility between mice with primary infection and secondary infection. PMID- 16696346 TI - Viral resistance: deja vu? PMID- 16696347 TI - Oval window fistula. PMID- 16696348 TI - An unusual cause of nasal obstruction: a hair clip in the nasopharynx. PMID- 16696349 TI - Vocal fold atrophy after resection of a parapharyngeal space sarcoma. PMID- 16696350 TI - Epithelial-myoepithelial carcinoma. PMID- 16696351 TI - Otogenic cerebellar abscess: an unusual occurrence. PMID- 16696352 TI - Early detection of gastric cancer with esophageal extension by transnasal esophagoscopy. PMID- 16696353 TI - Dizziness, hyperactive caloric responses, otic capsule demineralization, impaired glucose tolerance, and hyperinsulinemia. PMID- 16696354 TI - Web portals. PMID- 16696355 TI - Styloid-carotid artery syndrome. PMID- 16696356 TI - Barium esophagogram of a Zenker's diverticulopexy. PMID- 16696357 TI - Tinnitus and brain activation: insights from transcranial magnetic stimulation. AB - The mechanisms underlying tinnitus are still not completely elucidated, but advances in neuroimaging and brain stimulation have provided us with new insights. Evidence suggests that tinnitus might actually be generated by central rather than peripheral structures. To illustrate the importance of brain activity changes in the pathology of tinnitus, we report the cases of 2 patients who experienced a recurrence/worsening of their tinnitus after they had undergone treatment for major depression with repetitive transcranial magnetic stimulation. We suggest that the tinnitus in these 2 patients was induced by changes in brain activity resulting from transcranial magnetic stimulation of the prefrontal cortex. We also review the pathophysiology and other aspects of tinnitus, focusing on associated brain reorganization. PMID- 16696358 TI - Noise-induced perilymph fistula. AB - Perilymph fistulae are difficult to diagnose because they present with a wide variety of signs and symptoms, they are associated with many etiologies, and they often mimic other conditions. In this article, we describe a case of perilymph fistula that featured one of its more rare causes: acoustic trauma--specifically, damage from a loud blast from the siren of a fire engine. We also review the literature and discuss the difficulties of diagnosis and treatment and the possible mechanisms by which acoustic trauma and other etiologies cause perilymph fistulae. PMID- 16696359 TI - Underlay tympanoplasty with laser tissue welding. AB - We investigated the feasibility of using laser tissue welding techniques to perform transcanal underlay tympanoplasty. We used 10 temporal bones obtained from human cadavers. After creating a subtotal tympanic membrane perforation, we introduced harvested periosteum through the perforation and used laser tissue welding to secure the periosteum graft in place in an underlay fashion. The procedure was performed via a transcanal approach and did not require middle ear packing. Immediately after the graft had been placed, we qualitatively tested its integrity with a blunt probe. The graft was as strong as the native cadaver tympanic membrane in all 10 cases. We conclude that laser transcanal underlay tympanoplasty is a feasible and effective method of repairing a tympanic membrane. The ultimate goal is to develop a technique that will allow physicians to routinely perform underlay tympanoplasty on moderately sized perforations in an office setting. PMID- 16696360 TI - The mechanical reduction of early acquired cholesteatomas in children: indications and limitations. AB - The standard treatment for acquired cholesteatoma involves surgical removal of the lesion and reconstruction of the tympanic membrane. In some children, these lesions can be treated more conservatively. We conducted a retrospective study of 29 ears in 24 children who had been treated for early acquired cholesteatoma with mechanical reduction and a tympanostomy tube. Outcomes measures included hearing status, the postoperative appearance of the tympanic membrane, and the need for additional surgery. We found that anterior and inferior pars tensa lesions, with or without squamous debris, can be successfully reduced, but that posterosuperior retractions respond less well when the ossicular chain has been eroded. None of the children who responded to mechanical reduction required major reconstructive surgery later. We conclude that mechanical reduction of retraction pocket cholesteatomas with tympanostomy tube placement is sufficient to restore normal hearing and a normal tympanic membrane appearance in selected children with early lesions. We also identified several important prognostic features, including the patient's age, the specific location of the retraction pocket on the tympanic membrane, the extent of the pocket, ossicular chain involvement, and the patient's adenoid status. PMID- 16696361 TI - Postauricular abscess leading to internal jugular vein thrombosis: a case report. AB - Thrombosis of the internal jugular vein (IJV) is uncommon. It usually responds to conservative medical treatment. We describe a case of IJV thrombosis in a 31-year old man. The patient was successfully treated with IJV ligation after conservative therapy had failed. PMID- 16696362 TI - Outcomes of primary and secondary tracheoesophageal puncture: a 16-year retrospective analysis. AB - The current study retrospectively reviewed the cases of 68 patients who had undergone total laryngectomy and tracheoesophageal puncture (TEP) over a 16-year period. Fifty-one patients underwent primary TEP and 17 underwent secondary TEP. Nearly 80% of patients who received TEP at the time of laryngectomy achieved excellent voice quality perceptually. In contrast, only 50% of secondary TEP patients achieved excellent voice ratings. This difference was statistically robust (p = 0.03). Although both surgical and prosthesis-related complications occurred more frequently following primary TEP, statistically significant differences were not achieved. Neither pre- nor postoperative radiotherapy had any effect on voice restoration or complication rates. Based on these data, primary TEP may be preferable for several reasons, including a greater likelihood of successful voice restoration, a shorter duration of postoperative aphonia, and the elimination of the need for a second operation and interim tube feedings. PMID- 16696363 TI - Malignant melanoma of the mucous membranes of the head and neck: three case reports. AB - Primary malignant melanoma of the oronasal region is rare. When it does occur, it is difficult to manage, and the prognosis is not good. Early diagnosis and radical surgical management appear to offer the best hope of curing the disease. We report 3 new cases of malignant melanoma of the mucous membranes of the head and neck. Despite the generally poor outlook, 2 of these patients underwent excision of the tumor and remained disease-free 2 years later; the other patient refused surgery and was lost to follow-up. PMID- 16696364 TI - Management of a tracheal tear during laryngopharyngoesophagectomy with gastric pull-up. AB - Laceration of the posterior tracheal wall is one of the risks of transhiatal esophagectomy. Various methods of repairing such lacerations have been described; many of these methods involve a thoracotomy, but some do not. We describe a case of a posterior tracheal wall tear that occurred during a laryngopharyngectomy with a gastric pull-up. The tear was repaired with the transposed stomach and did not require a thoracotomy. The transposed stomach was used to patch the tear and block communication between the environment and the mediastinum. Bedside endoscopic examination on postoperative day 5 revealed that the tear had healed. Key management considerations in such a circumstance include having the patient breathe without positive pressure ventilation postoperatively and keeping the tracheal lumen and stoma clear during the healing process in order to prevent the development of positive tracheal pressure. With these safeguards in place, the transposed stomach approach is a safe method of repairing posterior tracheal wall tears. PMID- 16696365 TI - Primary malignant melanoma of the epiglottis: a rare presentation. AB - Primary malignant melanoma of the epiglottis is extremely rare. Until now, only 4 cases have been reported in the world literature. We describe a new case of epiglottic primary malignant melanoma in a 74-year-old man who presented with hoarseness and a foreign-body sensation. Clinical examination revealed the presence of a small, whitish, polypoid tumor on the laryngeal surface of the epiglottis; no other primary melanoma was detected. Wide excision of the lesion was performed, and microscopy revealed that it contained melanin-pigmented tumor cells in both the mucosa and submucosa. Immunostaining was positive for S-100 protein. The patient was treated with radiotherapy, and he remained well 1 year after the diagnosis with no evidence of recurrence. PMID- 16696366 TI - Factors that predict the need for intubation in patients with smoke inhalation injury. AB - Early identification of smoke inhalation patients who will require intubation is crucial. We conducted a retrospective chart review to identify predictors of respiratory distress in patients who present with smoke inhalation injury. Our study involved 41 patients who had been treated in the emergency room at a regional burn center. Eight of these patients required intubation. Intubation was positively correlated with physical examination findings of soot in the oral cavity (p < 0.001), facial burns (p = 0.025), and body burns (p = 0.025). The need for intubation was also predicted by fiberoptic laryngoscopic findings of edema of either the true vocal folds (p < 0.001) or the false vocal folds (p < 0.01). No statistically significant correlation was found between intubation and any of the classic symptoms of smoke inhalation: stridor, hoarseness, drooling, and dysphagia (all p = 1.0). Also, multivariate analysis revealed that facial burns correlated significantly with edema of the true vocal folds (p = 0.01) and body burns correlated significantly with edema of both the true (p = 0.047) and false (p = 0.003) vocal folds. We conclude that patients with soot in the oral cavity, facial burns, and/or body burns should be monitored closely because these findings indicate a higher likelihood of laryngeal edema and the need for intubation. PMID- 16696367 TI - Recent developments and future trends in nuclear medicine instrumentation. AB - Molecular imaging using high-resolution single-photon emission computed tomography (SPECT) and positron emission tomography (PET) has advanced elegantly and has steadily gained importance in the clinical and research arenas. Continuous efforts to integrate recent research findings for the design of different geometries and various detector technologies of SPECT and PET cameras have become the goal of both the academic comcameras have become the goal of both the academic community and nuclear medicine industry. As PET has recently become of more interest for clinical practice, several different design trends seem to have developed. Systems are being designed for "low cost" clinical applications, very high-resolution research applications (including small-animal imaging), and just about everywhere in-between. The development of dual-modality imaging systems has revolutionized the practice of nuclear medicine. The major advantage being that SPECT/PET data are intrinsically aligned to anatomical information from the X-ray computed tomography (CT), without the use of external markers or internal landmarks. On the other hand, combining PET with Magnetic Resonance Imaging (MRI) technology is scientifically more challenging owing to the strong magnetic fields. Nevertheless, significant progress has been made resulting in the design of a prototype small animal PET scanner coupled to three multichannel photomultipliers via optical fibers, so that the PET detector can be operated within a conventional MR system. Thus, many different design paths are being pursued--which ones are likely to be the main stream of future commercial systems? It will be interesting, indeed, to see which technologies become the most popular in the future. This paper briefly summarizes state-of-the art developments in nuclear medicine instrumentation. Future prospects will also be discussed. PMID- 16696368 TI - [Image reconstruction and quantification in emission tomography]. AB - Radionuclide tomographic imaging has substantially benefited from the introduction of statistical image reconstruction. Although the main concepts of these iterative algorithms were published decades ago, their widespread use in clinical routine only became available with faster computers for image processing in the last few years. This article gives an overview of data acquisition and iterative reconstruction in emission tomography, deals with the popular maximum likelihood algorithm, and describes the basics of the maximum a posteriori reconstruction. Prerequisites and corrections necessary for quantification are discussed in the second part of the article on the basis of positron emission tomography. Improvements in technical equipment are expected to stimulate future research into image reconstruction. PMID- 16696369 TI - From 2D PET to 3D PET: issues of data representation and image reconstruction. AB - Positron emission tomography (PET), intrinsically a 3D imaging technique, was for a long time exclusively operated in 2D mode, using septa to shield the detectors from photons emitted obliquely to the detector planes. However, the use of septa results in a considerable loss of sensitivity. From the late 1980s, significant efforts have been made to develop a methodology for the acquisition and reconstruction of 3D PET data. This paper focuses on the differences between data acquisition in 2D and 3D mode, especially in terms of data set sizes and representation. Although the real time data acquisition aspect in 3D has been mostly solved in modern PET scanner systems, there still remain questions on how to represent and how to make best use of the information contained in the acquired data sets. Data representation methods, such as list-mode and matrix based methods, possibly with additional compression, will be discussed. Moving from 2D to 3D PET has major implications on the way these data are reconstructed to images. Two fundamentally different approaches exist, the analytical one and the iterative one. Both, at different expenses, can be extended to directly handle 3D data sets. Either way the computational burden increases heavily compared to 2D reconstruction. One possibility to benefit from the increased sensitivity in 3D PET while sticking to high-performance 2D reconstruction algorithms is to rebin 3D into 2D data sets. The value of data rebinning will be explored. An ever increasing computing power and the concept of distributed or parallel computing have made direct 3D reconstruction feasible. Following a short review of reconstruction methods and their extensions to 3D, we focus on numerical aspects that improve reconstruction performance, which is especially important in solving large equation systems in 3D iterative reconstruction. Finally exemplary results are shown to review the properties of the discussed algorithms. This paper concludes with an overview on future trends in data representation and reconstruction. PMID- 16696370 TI - The impact of PET and SPECT on dosimetry for targeted radionuclide therapy. AB - Targeted radionuclide therapy (TRT) is an increasingly used treatment modality for a range of cancers. To date, few treatments have involved the use of dosimetry either to plan treatment or to retrospectively ascertain the absorbed dose delivered during treatment. Also the correlation between absorbed dose and biological effect has been difficult to establish. Tomographic methods permit the determination of the activity volume on a macroscopic scale at different time points. Proper attenuation correction in tomographic imaging requires a patient specific attenuation map. This can be obtained from scintillation-camera transmission scanning, CT or by using segmented scatter-emission images. Attenuation corrections can be performed either on the projection images, on the reconstructed images, or as part of an iterative reconstruction method. The problem of image quantification for therapy radionuclides, particularly for I 131, is exacerbated by the fact that most cameras are optimised for diagnostic imaging with Tc-99m. In addition, problems may arise when high activities are to be measured due to count losses and mis-positioned events, because of insufficient pile-up and dead time correction methods. Sufficient image quantification, however is only possible if all effects that degrade the quantitative content of the image have been corrected for. Monte Carlo simulations are an appealing tool that can help to model interactions occurring in the patient or in the detector system. This is helpful to develop and test correction techniques, or to help to define detectors better suited to quantitative imaging. PET is probably the most accurate imaging method for the determination of activity concentrations in tissue. PET imaging can be considered for pre-therapeutic treatment planning but ideally requires the use of a radioisotope from the same element as that used for treatment (e.g. I-124 for I 131; Y-86 for Y-90). Problems, however are that--some of the positron emitting isotopes have a shorter half-life--non-standard quantification procedures have to be performed--the availability of the radiopharmaceutical is presently limited; Many 3D-tools and -techniques are now available to the physicist and clinician to enable absorbed dose calculations to both target and critical organs-at-risk. The challenge now facing nuclear medicine is to enable this methodology to be routinely available to the clinic, to ensure common standard operating procedures between centres and in particular to correlate response criteria with absorbed dose estimates. PMID- 16696371 TI - Gated listmode acquisition with the quadHIDAC animal PET to image mouse hearts. AB - PURPOSE: The aim of this study was to develop ECG and respiratory gating in combination with listmode acquisition for the quadHIDAC small-animal PET scanner. METHODS: ECG and respiratory gating was realized with the help of an external trigger device (BioVET) synchronized with the listmode acquisition. Listmode data of a mouse acquisition (injected with 6.5 MBq of 18F-FDG) were sorted according to three different gating definitions: 12 cardiac gates, 8 respiratory gates and a combination of 8 cardiac and 8 respiratory gates. Images were reconstructed with filtered back-projection (ramp filter), and parameters like left ventricular wall thickness (WT), wall-to-wall separation (WS) and blood to myocardium activity ratios (BMR) were calculated. RESULTS: Cardiac gated images show improvement of all parameters (WT 2.6 mm, WS 4.1 mm, BRM 2.3) in diastole compared to ungated images (WT 3.0 mm, WS 3.4 mm, BMR 1.3). Respiratory gating had little effect on calculated parameters. CONCLUSION: ECG gating with the quadHIDAC can improve myocardial image quality in mice. This could have a major impact on the calculation of an image-derived input function for kinetic modelling. PMID- 16696372 TI - Concepts of registration and correction of head motion in positron emission tomography. AB - The long acquisition times (up to hours) in PET brain imaging bear a high risk of head motion, which results in artefacts like blurred images and may even lead to misinterpretation and useless data. With the increased resolution of high performance PET scanners, the influence of head movements becomes more and more relevant. Especially in the analysis of small brain structures, e.g. during ROI analysis, head motion results in inaccuracies of quantified data. This may also influence the kinetic analysis and generate artifacts in parametric images calculated from a motion-affected image sequence. This work presents the feasibility of head motion registration using an external motion tracking system. The implementation of the multi acquisition frame method and an event-by-event method to correct PET data for motion are described. The effects of motion correction are demonstrated on the basis of phantom measurements and patient data. The influence of motion correction on parametric imaging is described in a receptor study. PMID- 16696373 TI - Optimized list-mode acquisition and data processing procedures for ACS2 based PET systems. AB - PET systems using the acquisition control system version 2 (ACS2), e.g. the ECAT Exact HR PET scanner series, offer a rather restricted list-mode functionality. For instance, typical transfers of acquisition data consume a considerable amount of time. This represents a severe obstacle to the utilization of potential advantages of list-mode acquisition. In our study, we have developed hardware and software solutions which do not only allow for the integration of list-mode into routine procedures, but also improve the overall runtime stability of the system. We show that our methods are able to speed up the transfer of the acquired data to the image reconstruction and processing workstations by a factor of up to 140. We discuss how this improvement allows for the integration of list-mode-based post-processing methods such as an event-driven movement correction into the data processing environment, and how list-mode is able to improve the overall flexibility of PET investigations in general. Furthermore, we show that our methods are also attractive for conventional histogram-mode acquisition, due to the improved stability of the ACS2 system. PMID- 16696374 TI - Performance evaluation of the PET sub-systems of two PET/CT imagers according to NEMA standard NU 2-2001. AB - Performance and image quality of PET scanners are mainly determined by the choice of the detector material and the realised design of the scanner. The aim of this study was to measure the PET performance characteristics of two state-of-the-art PET/CT Systems. METHODS: Performance measurements following the NEMA NU 2-2001 standard were performed on a GSO-based system and on a system with LSO detectors. The following parameters were determined: spatial resolution, scatter fraction, sensitivity and count rate behaviour A body phantom was used to determine the image quality as well as the influence of differently performed attenuation corrections (low dose CT vs. 137Cs transmission source). RESULTS: Both scanners have partly different, but altogether good performance characteristics. The differences can be explained mostly by different physical properties of the detectors and by a different design of the scanners including the implemented electronics. CONCLUSION: If the data are to be compared quantitatively, the modality of the attenuation correction has to be taken into account, especially in regions with reduced uptake or reduced density. Both scanners achieve a very good image quality rich in contrast within short acquisition times which allows easy detection of small lesions. PMID- 16696375 TI - Organ movement reduction in PET/CT using dual-gated list-mode acquisition. AB - PURPOSE: Imaging of moving organs using the PET leads to blurred images due to long acquisition times. Simultaneous cardiac and respiratory gating of list-mode PET/CT is evaluated with the aim to improve image quality and assess the organ movement. METHODS: We performed a N-13 ammonia PET/CT scan with a human volunteer, using the Siemens Biograph Sensation 16 scanner with list-mode acquisition. For ECG gating we used the scanner's integrated ECG device. Respiratory gating was done with the BioVet pneumatic sensor system. RESULTS: The sorting of the list-mode data post acquisition produced the desired matrix of eight cardiac times eight respiratory images. Organ movement could be measured in the series of gated PET images. The quantitation of tracer uptake in the myocardium showed artifacts due to the CT-based attenuation correction. CONCLUSION: Double gating is feasible in human PET/CT scans using a list-mode based scan protocol. The image quality can be enhanced using double gated list mode acquisition in PET/CT Attenuation correction protocols in PET using a single not gated fast CT introduces artifacts in moving organs. PMID- 16696376 TI - [Chromosomal disorders in the background of azoospermia]. AB - INTRODUCTION: Nowadays more and more couples face the fact that they cannot have babies in spite of many years of trying. The male factor can be identified in about half of these cases. AIM: The aim of this study was to analyse chromosomal alterations in patients with azoospermia. These patients may be candidates for testicular sperm retrieval and intracytoplasmatic sperm injection. MATERIALS AND METHODS: Preoperative evaluation included routine andrological investigation with 2 semen analysis, ultrasound, hormonal and genetic examination. Traditional histological examination and embryological diagnostic of tissue samples was performed. Cryopreservation of retrieved testicular tissue was also done. Between January 2001 and June 2005 73 biopsies were performed in 71 patients for testicular sperm extraction. In order to obtain an exact diagnosis, the traditional cytogenetic methods and fluorescence in situ hybridization analyses were performed in combination with molecular genetic techniques. Patients were offered to participate in the assisted reproduction programme on the base of their genetic results. RESULTS: In this study, the most characteristic cases were numerical deviations, such as 47,XXY (2 cases), mosaic 47,XXY/49,XXXXY (1 case), 47,XYY (1 case) and mosaic 46,XY/45,X (1 case) karyotypes. Non-obstructive azoospermia was diagnosed in 53 patients (79%). CONCLUSIONS: Authors emphasised the importance of cyto- and molecular examinations in cases of genetical disorders. The results provide a chance for patients to be spared from long-drawn moreover psychological burdening examinations. In addition the costs of different clinical intervention could be saved too. PMID- 16696377 TI - [Wertheim-operation: 5-year survival of 501 consecutive patients with cervical cancer]. AB - OBJECTIVE: The purpose of this study was to assess the 5-year survival and morbidity when radical hysterectomy and pelvic lymphadenectomy with pre- and postoperative radiation have been performed for IA2-IIB stage cervical cancer. STUDY DESIGN: During a 10 and a half year period between July, 1990 and December, 2000, 501 consecutive radical hysterectomies with bilateral pelvic lymphadenectomies were performed by the same gynecological surgeon for stage IA2, IB, IIA and IIB carcinoma of the cervix at the I. Department of Obstetrics and Gynecology, Semmelweis University Budapest. The patients were treated by pre- and postoperative irradiation as well. RESULTS: Perioperative complications apart from recurrence were minimal with no long-term morbidity. The absolute 5-year survival rates for the patients in stage IA2, IB1, IB2, IIA and IIB were 94,4%, 90,7%, 84,1%, 71,1% and 55,4%, respectively. The respective 5-year survival rates for patients without or with lymph node metastasis, were 94,5% and 33,3% in stage IB2, 81,7% and 48,7% in stage IIA and 70,2% and 36,5% in stage IIB, respectively. CONCLUSION: Nerve-sparing radical abdominal hysterectomy with pelvic lymph node dissection and pre- and postoperative irradiation remains the treatment of choice for most patients with early-stage and even in IIB-stage cervical cancer. The radicality and extent of lymph node dissection and parametrial resection should be individualized and tailored to tumor- and patient-related risk factors. PMID- 16696378 TI - [Association of chronic hepatitis C and lichen planus--in two patients]. AB - Relationship between hepatitis C virus (HCV) infection and lichen planus (LP) is well known but controversial in the literature. AIM: Two patients with lichen planus and chronic hepatitis C are presented. One of them suffered with disseminated LP and the other had cutaneous and oral erosive symptoms. METHODS: The chronic hepatitis of the patients was proved by elevated ALT and AST level, anti HCV (ELISA) and HCV-PCR serological examinations and liver biopsy. The diagnosis of lichen planus established on he typical clinical symptoms and the histological examination. RESULTS: HCV RNA in the skin specimen from the biopsy of the skin lesion was detected by RT-PCR method, but the non affected skin specimen from the first patient was HCV RNA negative. Treatment of the chronic hepatitis C with specific therapy (interferon-alpha + ribavirin) in the second patient led to the regression of lichen planus symptoms. CONCLUSIONS: The authors supposed that the lichen planus is one of the extrahepatic manifestations of HCV infection and there is a real correlation between the two diseases in these cases. PMID- 16696379 TI - [Clinical features and pathogenesis of Carney complex, a rare form of multiple endocrine neoplasia syndromes]. AB - Carney-complex is the rarest and most recently described form of multiple endocrine neoplasia syndromes that is unique both from clinical and pathogenetic aspects. Clinical features include spotty skin pigmentation, cutaneous and cardiac myxomas, multiple endocrine abnormalities and schwannomas. The most characteristic endocrine abnormality is primary pigmented nodular adrenal hyperplasia that may result in clinically apparent Cushing's syndrome. Acromegaly, hyperprolactinaemia, tumours of the testis and ovaries have also been described. Approximately fifty percent of Carney-complex cases are familial, with an autosomal dominant inheritance pattern. About 45-65% of Carney-complex cases are related to mutations of the PRKAR1A gene encoding a regulatory subunit of protein kinase A, but other genetic mechanisms seem to be involved, as well. Here, the authors present a brief synopsis of its clinical and pathogenetical features. PMID- 16696380 TI - [Professor Ferenc Kovacs, second director of the Department of Obstetrics and Gynecology of Debrecen University]. PMID- 16696381 TI - [Reminiscence of Gyorgy Hertz Dan (1922-1998)]. PMID- 16696382 TI - The prevalence of streptococcus agalactiae (group B) colonization in pregnant women at Thammasat Hospital. AB - BACKGROUND: Group B Streptococci (GBS) is responsible for serious infections in newborns such as septicemia and meningitis. OBJECTIVE: The present study was carried out to find the prevalence of GBS colonization in pregnant women and to determine the pattern of antibiotic resistance of the isolates. MATERIAL AND METHOD: From November 2004 to February 2005, 406 pregnant women were examined for GBS. RESULTS: GBS colonization rate was 16% in pregnant women, receiving antenatal care at Thammasat Hospital. CONCLUSION: All the isolates were sensitive to Ampicillin, Penicillin, Vancomycin and Cephazolin. Resistant was seen with Clindamycin (3%) and Erythromicin (1.5%). PMID- 16696383 TI - CNS germ cell tumors: pattern of failure and effects of radiation volume. AB - This retrospective study was conducted to evaluate local control and overall survival after radiotherapy for patients with intracranial germ cell tumors and to investigate the influence of irradiated field on treatment outcome. Thirty-two patients with surgically confirmed or suspected primary intracranial germ cell tumors (GCT) treated at the Division of Therapeutic Radiology and Oncology, Chiang Mai University, Chiang Mai, Thailand between January 1988 and December 1999 were reviewed Seven patients were not included in the analysis of treatment outcome and survival due to incompleteness of radiation treatment or death before the end of treatment. The median follow up time of 39.5 months (range from 2.3 months to 136.1 months). Median age at diagnosis was 16.5 years with 23 males and 9 females. Patients were irradiated to the primary tumor with an adequate margin in 7 patients, to the whole brain with a cone down boost in 8 patients. Craniospinal irradiation (CSI) was performed in 10 patients. For the 25 evaluable patients, 5 year overall survival was 86.4%. Five-year disease free survival was 72.9%. Five year overall survival rates were 83.1% and 90.0% for the germinoma and nonbiopsied group. (p-value = 0. 6052). Routine prophylactic CSI was not given with a spinal only failure rate of 33.3%. Five-year overall survival were 85.7%, 87.5%, 85.7% for CSI, whole brain irradiation with boost and local field irradiation (p-value = 0.9037). Five-year disease free survival were 85.7%, 72.9%, 85.7% for CSI, WBRT, and local field (p-value = 0. 6403). This retrospective study suggests that definitive radiation therapy is effective in controlling germinoma, and cure rates are excellent with irradiation alone. Craniospinal irradiation can eliminate the risk of relapse especially in patients who had incomplete diagnostic craniospinal evaluation. PMID- 16696384 TI - MRI of multiple sclerosis in Thai patients. AB - OBJECTIVE: Evaluate the MRI findings of multiple sclerosis (MS) in Thai patients and compare these with the McDonald criteria for dissemination in space. MATERIAL AND METHOD: A retrospective study was performed by reviewing clinical and MRI records of patients attending MS clinic at Siriraj Hospital. Only patients with complete clinical and MRI data were included in the present study. RESULTS: There were 25 patients included in the study. Twenty-four patients were females and 1 was male. The mean age was 35.24 years old (range 16-50). The disease duration was 1 month to 17 years after the onset. Twenty-three patients were diagnosed as relapsing-remitting MS, one as possible MS and one as clinical isolated syndrome. Analysis for CSF oligoclonal bands was performed in 22 cases with positive results in 6 cases (27.3%). The sites of involvement were optic neuritis (16 cases), spinal cord (14 cases), and brain (9 cases). Seventeen cases had MRI study at the early presentations, and 8 cases during subsequent relapses. In the 9 cases with brain symptoms, all had positive brain MRI. Ten cases (58.8%) had infratentorial lesions mostly found at the medulla. Most T2 lesions were found at juxtacortical and periventricular regions. Five in 9 cases (55.6%) with periventricular lesions had lesions not less than 3 mm and at least 3 lesions were found. There were 6 cases (35.3%) with, at least, nine T2 lesions. Most of the T2 lesions were oval shaped. Seven cases (41.2%) had black hole lesions. Three cases (17.6%) had gadolinium enhanced lesions. Thirteen of 15 available spinal MRI were abnormal. The locations of the lesions were cervical (6), thoracic (4) and cervicothoracic (3) levels. The number of T2 lesions was one in 8 cases and more than one in 5 cases. The length of each lesion varied: less than 1 vertebral segment (5 cases) and 2 or more vertebral segments (8 cases). Swelling of the cord was found in 3 cases and atrophy in 7 cases. The lesions occupied the whole cross-sectional cord in 6 cases and center of the cord in 7 cases. The gadolinium enhancement was found in 6 cases with a patchy pattern in 5 cases and mixed ring and patchy patterns in 1 case. There were totally 6 cases (24%) that fulfilled the McDonald MRI criteria for dissemination in space. CONCLUSION: The MRI findings in Thai MS have distinct features from the Western reports. Re-evaluation of McDonald criteria for the Eastern countries is needed. PMID- 16696385 TI - Postoperative radiotherapy with high dose rate iridium 192 mould for prevention of earlobe keloids. AB - BACKGROUND: A 50% to 80% recurrence rate of earlobe keloids can occur following a simple excision. Many modalities, including radiotherapy, have been suggested to reduce the postoperation recurrence. OBJECTIVE: The aim of the present study was to determine the efficacy of postoperative radiotherapy by a high dose rate Iridium 192 mould in the prevention of earlobe keloids recurrence. MATERIAL AND METHOD: Between March 1999 and March 2003, 22 patients with 24 earlobe keloids were treated by radiotherapy immediately following surgical excision. A dose of 15 Gray in 3 fractions was delivered at a point placed 5 mm from the axis of the Iridium sources. RESULTS: From the 22 patients with 24 keloids who were treated, 15 patients with 16 keloids were followed for a minimum of 6 months. Recurrence occurred in two keloids (12.5%). There were no severe adverse effects. CONCLUSION: Postoperative radiotherapy by high dose rate Iridium 192 mould was an effective prevention of earlobe keloids recurrence. It was well tolerated and did not present any significant side effects. PMID- 16696386 TI - Predictive CT features in ruptured abdominal aortic aneurysm. AB - OBJECTIVE: To evaluate the CT findings including size of aneurysm, hyperattenuation crescent sign and focal discontinuity of mural calcification to predict the risk of ruptured aneurysm. MATERIAL AND METHOD: Records of 27 patients, who underwent Multislices Computed Tomography (MDCT) and required operative repair of abdominal aortic aneurysm from July 2000 to July 2003 were retrospectively reviewed Two radiologists evaluated the images by consensus, analyzing the aneurysm size, hyperattenuation crescent sign, and focal discontinuity of mural calcification. RESULTS: The authors found that the mean maximum AP diameters of the aneurysm in the ruptured and nonruptured group were 6.95 cm and 5.50 cm, respectively. All patients in the ruptured group had an aneurysm size of more than 5.0 cm. The hyperattenuation crescent sign and focal discontinuity of mural calcification had a high specificity for predicting ruptured aneurysm, 95% and 100%, respectively. There was statistical significant difference between the ruptured aneurysm and non-ruptured groups in the patients who had maximum aneurysm size more than 5 cm with positive hyperattenuation crescent sign (p < 0.041). CONCLUSION: A maximum size of aneurysm greater than 5 cm with positive hyperattenuation crescent sign is a suggestive sign to predict ruptured aneurysm. PMID- 16696387 TI - A 12-case series of Penicillium marneffei pneumonia. AB - BACKGROUND: Penicillium marneffei, an endemic fungus in Southeast Asia and southern China, is the cause of opportunistic infection in HIV-infected patients who may present with symptoms and signs of the lungs, and abnormal chest radiographs. However, only a few cases of pulmonary infection from this organism have been reported. OBJECTIVE: To study the clinical manifestations of patients with Penicillium marneffei pneumonia diagnosed by sputum or bronchoalveolar lavage (BAL) fluid culture MATERIAL AND METHOD: Retrospective descriptive study of patients who were diagnosed with Penicillium marneffei pneumonia at Maharaj Nakorn Chiang Mai Hospital from September 1999 to July 2004. RESULTS: Twelve patients (eight males, four females) were included with mean age of 36.1 years. Nine cases were HIV-infected. Their presenting symptoms included fever, cough, dyspnea and weight loss. Skin lesions, hepatomegaly and lymphadenopathy were extrapulmonary signs. Chest radiographs revealed diffuse reticulonodular, diffuse reticular, localized alveolar, localized reticular infiltration, and cavitary lesion. The diagnosis was made by cultures from the sputum in five cases and BAL fluid in the others. Co-infections with Streptococcus pneumoniae, Klebsiella pneumoniae, Mycobacterium tuberculosis, Cryptococcus neoformans, and Strongyloides stercoralis were found. Most of them were treated by intravenous amphotericin B followed by oral itraconazole, or oral itraconazole. CONCLUSION: Penicillium marneffei pneumonia has non-specific clinical manifestations, it cannot be excluded from other infections and may have co-infections. Physicians should include this infection in their differential diagnosis especially in immunocompromised patients. PMID- 16696388 TI - Validation of the Thai version of a screening questionnaire for detection of systemic lupus erythematosus. AB - OBJECTIVE: The present study was performed to assess the sensitivity and specificity of the translation questionnaire for Systemic Lupus Erythematosus (SLE) in Thai speaking populations. MATERIAL AND METHOD: The 10- item questionnaire was applied to out-patients at the rheumatology clinic of the Chiang Mai University Hospital. One hundred and thirty-nine SLE, 109 Rheumatoid Arthritis (RA), and 35 Scleroderma (Scl) patients, as well as 88 Healthy Controls (HC) were enrolled into the present study. RESULTS: All subjects completed the questionnaire within 2 minutes. A positive response to three or more questions of the questionnaire gave a sensitivity and specificity of 92.81% and 76. 39%, respectively, and was comparable to the original version. CONCLUSION: This Thai version of the screening questionnaire should be applied in the general population to determine the prevalence of SLE. PMID- 16696389 TI - Closed vs ligasure hemorrhoidectomy: a prospective, randomized clinical trial. AB - OBJECTIVE: To compare the operative time, postoperative complications, and analgesic requirement between closed hemorrhoidectomy and Ligasure hemorrhoidectomy. MATERIAL AND METHOD: The study was conducted in a prospectively randomized controlled fashion. Forty-seven patients with grade 3 or 4 hemorrhoids plus external component or skin tag were operated on by either hemorrhoidectomy with Ligasure (24 patients) or closed hemorrhoidectomy (23 patients). One patient in each group was lost to follow up. The operative time, postoperative verbal numeric pain score, analgesic requirement, bleeding, and wound dehiscence between the two groups were compared Unpaired t-tests, Mann-Whitney U tests, or Fisher's Exact tests were used where appropriate. RESULTS: Demographic and clinical data between two groups were comparable. Operative time for the Ligasure hemorrhoidectomy was significantly shorter than the closed hemorrhoidectomy group (21.70 +/- 11.76 vs 35.68 +/- 14.25 min, p < 0. 001), while the number of resected hemorrhoids in the study group were 2.91 versus 2.18 in the control group. However, there were no differences in post-operative pain score, analgesic requirement, bleeding, or wound dehiscence between the two groups. CONCLUSION: Ligasure hemorrhoidectomy is superior to closed hemorrhoidectomy in terms of reducing the operative time without affecting postoperative complications. PMID- 16696390 TI - Management of endobronchial cancer using bronchoscopic electrocautery. AB - From December 2004 until November 2005, 15 cases of endobronchial cancer presenting with 22 episodes of mechanical obstruction of the airway, difficulty in breathing, hemoptysis, severe coughing, obstructive pneumonia and identified recurrent atelectasis of the lungs. The cancer was removed rapidly, effectively and without complication by a technique of fiberoptic bronchoscopic electrocautery. PMID- 16696391 TI - Postoperative analgesia for total knee replacement: comparing between pre-and postoperative "3-in-1" femoral nerve block. AB - BACKGROUND: Total Knee Replacement (TKR) produces severe postoperative pain. Pre- and postoperative single-shot "3-in-1" Femoral Nerve Block (FNB) were reported to improve analgesia and reduce morphine consumption post TKR. OBJECTIVE: To find out the most beneficial time for injection of single shot "3-in-1" FNB for TKR between preoperative and postoperative in a prospective controlled trial. MATERIAL AND METHOD: In a Randomized, double-blind Controlled Trial (RCT), 48 patients undergoing TKR received either pre- or postoperative "3-in-1" FNB using 30 mL of bupivacaine 0.25% after a standardized general anesthesia. Morphine consumption, Numeric Pain-Rating Scale (NPRS) at rest and during movement, tension in the back of the knee, nausea/vomiting, pruritus, sedation, and respiratory depression at 1, 4, 24 and 48 hr after TKR were compared RESULTS: There were no significant differences in 48-hr morphine consumption [46.5 (20.0) vs 45.0 (23.6) mg, p = 0.809], NPRS both at rest and during movement, tension in the back of knee, nausea/vomiting, pruritus, sedation, and respiratory depression at any time during 48-hr postoperative TKR between groups. CONCLUSION: Preoperative single-shot "3-in-1" FNB using 30 mL of bupivacaine 0.25% is not better than postoperative single-shot "3-in-1" FNB using the same drug in postoperative pain and morphine reduction in patients undergoing elective TKR under general anesthesia. PMID- 16696392 TI - Four recti enlargement at orbital apex and thyroid associated optic neuropathy. AB - OBJECTIVE: To determine the site of muscle enlargement in thyroid-associated ophthalmopathy and optic neuropathy in Ramathibodi Hospital. MATERIAL AND METHOD: Data and images of MRI and CT of patients with thyroid-associated ophthalmopathy were retrospectively reviewed Neuro-ophthalmologic data and correlation to the imaging findings were analyzed. RESULTS: Among 19 patients with thyroid associated ophthalmopathy, 20 imaging studies were collected. There were 4 patients with bilateral disease and crowdedness of 4 recti muscle. All of them had thyroid associated optic neuropathy. One out of 19 patients underwent two different sessions study. During her first study, right optic nerve was involved with the crowding of right optic nerve but 7 months later her left optic nerve became involved with crowding of the left optic nerve and the right 4 recti appeared smaller CONCLUSION: Four recti muscle enlargement in thyroid-associated ophthalmopathy, like four-leaf clover in coronal section of imaging study, may be an important sign of thyroid associated optic neuropathy. PMID- 16696393 TI - Validity and reliability study of the Thai version of WHO SCAN: somatoform and dissociative symptoms section. AB - OBJECTIVES: To determine the validity and reliability of the Thai version of the WHO Somatoform and Dissociative Symptoms Section of the Schedules for Clinical Assessment in Neuropsychiatry (SCAN) Version 2.1 MATERIAL AND METHOD: The SCAN interview version 2.1 Somatoform and Dissociative Symptoms Section was translated into Thai. The content validity of the translation was verified by comparing a back-translation (to English) of the Thai version to the English original. Whenever inconsistencies were encountered, the Thai version was adapted so that it correctly conveyed the meaning of the original English version. The revised Thai version was then field-tested nationwide for the comprehensibility of the relatively technical language. Between October 2003 and August 2004, 30 persons were recruited for the reliability study (16 males; 14 females) Fifteen subjects had somatoform disorders and 15 were normal. The number of years of formal education varied widely and occupations were diverse. Subjects were interviewed by a psychiatrist competent in using the Thai version of SCAN. The interviews were recorded on video so that the material could be rerated. RESULTS: Based on the response from Thai subjects and consultations with competent psychiatrists, the content validity was established. The time taken to interview a somatoform patient averaged 57.1 +/- 12.1 minutes while it was 42.1 +/- 13.9 minutes for a normal subject. The inter-rater reliability (kappa) of the 113 Items were: 0.81 1.0, 0.61-0.80 and 0. 00-0.20 in 49.6, 30.0 and 8.9 percent, respectively. Kappas could not be calculated for 11.5% of the Items. The intra-rater reliabilities were. 0.81-1.0, 0.61-0.80 and 0.00-0.20 in 54.9, 26.5 and 2.7 percent, respectively. Kappas could not be calculated for 15.9% of the Items. CONCLUSION: The Thai version of the Somatoform and Dissociative Symptoms Section of SCAN version 2.1 proved to be a valid and reliable tool for assessing somatoform and dissociative symptoms among Thai speakers. PMID- 16696394 TI - Association between different variants of papillary thyroid carcinoma and risk group according to AMES (age, metastasis, extent and size) classification system. AB - OBJECTIVE: To study the association between cell variants of papillary carcinoma and AMES (Age, Metastasis, Extent and Size) risk classification. MATERIAL AND METHOD: One hundred and twenty-one cases of papillary thyroid carcinomas were subclassified for cell type and risk-groups according to AMES classification system. Correlations between both variables are evaluated RESULTS: Among different cell variants of papillary carcinoma, solid cell pattern has the highest proportion of high-risk tumor classified by the AMES criteria, comprising 75% followed by tall cell subtype with 33.3% of high risk patients. Conventional papillary carcinoma has only 8.3% of high-risk group. Follicular and encapsulated variants as well as microcarcinoma (< 1 cm) are all categorized as low-risk neoplasms. CONCLUSION: The present study indicates that there is association between cell variants and AMES prognostic index. The authors, therefore, emphasize the importance of cell variants in predicting the prognosis of papillary carcinoma. PMID- 16696395 TI - Serum leptin concentrations in chronic hepatitis. AB - The objectives of this research were to investigate the leptin levels among Chronic Hepatitis B Virus (HBV), Chronic Hepatitis C Virus (HCV) and non alcoholic steatosis hepatitis (NASH) diseases of Thai patients compared with controls. Twenty of each HBV, HCV and NASH patients compared with sixty people as the control group from the Outpatient Department at the Hospital for Tropical Diseases, Bangkok, Thailand were investigated. Fasting blood samples were collected for investigation of leptin concentration, liver enzyme function tests and hematological variables. The serum leptin concentration of liver patients was significantly higher than that of control subjects. It might be due to the accumulations of fat cells in liver disease patients. However, there is no relationship between leptin level and other parameters such as BMI, ALT, AST, ALP and hematological variables. Liver enzyme functions levels are much higher in patients groups. White blood cells counts, platelets and hematocrit values are slightly lower in liver disease patients. Therefore, it is concluded that physiological regulation of leptin maintains in relation to body fat, even in chronic viral liver diseases. This finding and the apparent stage suggest the possibility that in the course of chronic viral diseases, serum leptin levels may reflect the extent of liver dysfunction. PMID- 16696396 TI - Development of community-based speech therapy model: for children with cleft lip/palate in northeast Thailand. AB - BACKGROUND: Surgical treatment can reduce disfigurement for children born with cleft lip/palate, however, most children are left with speech and language problems. This creates a new problem as speech and language services is limited. OBJECTIVE: To combine the principles of Community-Based Rehabilitation (CBR), Primary Health Care (PHC) and institutional medical approaches for reaching and treating speech disordered children with cleft lip and/ or palate in remote area. MATERIAL AND METHOD: The authors conducted the study from participatory workshops for development of a Community-Based Model. RESULTS: Community-Based Speech Therapy Model for children with cleft lip/palate was established based on healthcare system. CONCLUSION: Model can be implemented among children with cleft lip/palate for further process in Northeast and other areas of Thailand as well as developing countries where there is a limitation of speech therapy. PMID- 16696397 TI - An increase of the cardiothoracic ratio leads to a diagnosis of Bart's hydrops. AB - BACKGROUND: Thalassemia is a common single gene disorder in Southeast Asia. a thalassemia is a group of syndrome characterized by deficient production of the alpha-globin chain. Individuals with heterozygous alpha-thalassemia-1 are at risk of having a fetus that has Hemoglobin Bart's hydrops fetalis (Hb Bart's). Usually, when the hemoglobin electrophoresis in heterozygous alpha-thalassemia-1 is normal, the Mean Corpuscular Volume (MCV) is lowered. We report a case of increased cardiothoracic ratio that led to a diagnosis of Hb Bart's in a couple who had normal hemoglobin electrophoresis and low MCV. CASE REPORT: A 23-year-old woman, gravida 2, Para 0-0-1-0, initially presented for antenatal care at 13 weeks pregnancy. Her MCV was 67 fentolitre, DiChlorophenol-IndolPhenol (DCIP) test was negative and hemoglobin electrophoresis was normal. Her husbands MCV was 67 fentolitre, and hemoglobin electrophoresis was normal. Cardiomegaly (an increased of the cardiothoracic ratio) was detected by ultrasonogram at 25 weeks of gestation. She and her husband were comprehensively counseled after an Hb Bart's was suspected. A cordocentesis was performed and the fetal blood was tested for hemoglobin electrophoresis. The result was later known and confirmed as Hb Bart's. The couple decided to terminate the pregnancy. The induced abortion was successful and the patient was discharged on the second day after the abortion. She was well at the 4-week follow-up. CONCLUSION: A prenatal ultrasonographic screening should be conducted in couples who are suspected of being alpha-thalassemia-1 carriers when DNA study of alpha-globin gene cannot be performed. The increase of cardiothoracic ratio will help detect an early stage of Hb Bart's. PMID- 16696398 TI - Spontaneous rupture of an ovarian dermoid cyst associated with intra-abdominal chemical peritonitis: characteristic CT findings and literature review. AB - A case of ruptured ovarian dermoid is documented including the characteristic CT findings of chemical peritonitis based on the fatty peritoneal fluid content similar to that found in fatty dermoids. PMID- 16696399 TI - Respiratory complication of tsunami diaster victims in Vachira Phuket Hospital. AB - There were 958 patients who suffered from the tsunami diaster on 26 December 2004, and came to Vachira Phuket Hospital. 326 cases were admitted with severe respiratory complications. 42 cases had sputum culture, and 20 of them grew gram negative microorganism. The authors present 3 cases. PMID- 16696400 TI - Infective endocarditis in hypertrophic cardiomyopathy--mural and aortic valve vegetations: a case report. AB - Hypertrophic cardiomyopathy (HCM) is infrequently complicated by infective endocarditis (IE). The authors report the case of a 46-year-old woman developing IE in asymptomatic HCM. Blood cultures were positive for Streptococcus viridans. Echocardiography demonstrated: 1) a mobile (1.2 x 1 cm2) vegetation attached to the septal endocardium at the site of contact between the mitral valve leaflet and the hypertrophic septum; 2) two large (2.7 x 1.7 cm2 and 1.6 x 1.1 cm2) vegetations at NCC and RCC respectively of aortic valve, causing moderate valve regurgitation, and, 3) a mural (1 x 0.8 cm2) vegetation on the posterior wall of the left ventricle. On the third day of hospitalization, the patient underwent aortic valve replacement and removal of the vegetations. Antibiotics were continued for another four weeks. The patient recovered and follow-up was uneventful. Thus, chronic endocardial trauma of the septum, a common finding in HCM with outflow tract obstruction, may provide a fertile nidus for the development of vegetation, which in turn would play the major role in the pathogenesis of IE. PMID- 16696401 TI - The first invasive EEG monitoring for surgical treatment of epilepsy in Thailand. AB - The authors report the first invasive electroencephalography (EEG) monitoring in Thailand in a patient who had intractable left temporal lobe epilepsy. The seizure origin and functioning cortices were identified The corresponding epileptogenic zone was resected without functional deficit. The patient has become seizure free 1 year since surgery. Subdural EEG monitoring with cortical stimulation have been developed at this tertiary epilepsy center The technique provides essential evidence for the surgical decision so that the best post operative outcome can be achieved. PMID- 16696402 TI - Neuropathic arthropathy of the elbow: a report of two cases. AB - BACKGROUND: Neuropathic arthropathy (charcot joint) of the elbow is a rare condition. It is among the least frequently reported charcot pathology of the joint. The clinical symptoms are characterized by a painless and unstable joint. Infection in the neuropathic joint is rare. Only a limited number of cases have been reported in the literature. The authors present two cases of neuropathic arthropathy of the elbow. One of them was diagnosed as Septic neuropathic arthropathy. MATERIAL AND METHOD: Two patients with neuropathic arthropathy of the elbow (an infected and a noninfected case) were treated at our department. Both patients were over 60 years old. The underlying conditions associated with the arthropathy included combined median and ulnar neuropathy in the infected case and idiopathic in the non-infected case. Both of the patients had a history of specific trauma at the affected elbows. The radiographs of the elbows revealed dislocation, fracture fragmentation and some bony sclerosis. The management in the non-infected case aimed to maintain the function of the elbow with a short period of immobilization and physical therapy. For the infected case, the patient was treated successfully with systemic antibiotic, surgical debridement and a period of immobilization with an external fixator RESULTS: The patients were followed-up for approximately 9 months for the non-infected and 2 years for the infected case. Both of them had painless and functional range of motion of the elbows in the most recent follow-up examination. For the infected case, the surgical incision healed well and there was no recurrent discharge from that affected elbow. CONCLUSION: In the first case, with neuropathic arthropathy of the elbow, the investigations were made to find the underlying disease. The second case with septic neuropathic joint, systemic antibiotic, surgical debridement and appropriate immobilization were needed. With gross instability and marked distortion of the joint, the elbow of both patients functioned remarkably well. PMID- 16696403 TI - Chediak-Higashi syndrome: report of a case with uncommon presentation and review literature. AB - Chediak-Higashi syndrome (CHS) is a very rare autosomal recessive immunodeficiency disorder characterized by partial albinism, recurrent pyogenic infections, and large granules in all granule-containing cells. The author describes a Thai girl who was the first case of CHS in Thailand. She presented in the accelerated phase of CHS, which leads to repeated infections and bleeding, often resulting in fatal outcome. Pancytopenias, hepatosplenomegaly, lymphohistiocytic infiltration in bone marrow and the abnormal characteristic granules in leukocyte clinched the diagnosis. PMID- 16696404 TI - Paraurethral leiomyoma: a case report. AB - A paraurethral leiomyoma is a rare benign hormone-dependent tumor of mesenchymal origin. A 43-year-old woman presented with a palpable mass near the urethral opening with no abnormal urinary symptom. Simple excision was performed Histopathological diagnosis was paraurethral leiomyoma with confirmation by immunohistochemical study. Further hormonal study of the tumor yielded positive estrogen and progesterone receptor expression, suggesting conservative treatment with gonadotropin-releasing hormone (GnRH) agonist or antagonist may have a role as an alternative treatment in an inoperable case. PMID- 16696405 TI - Cell therapy: hype or hope. AB - Cell therapy is a promising therapeutic tool for the next decade. It has a potential to cure a number of chronic diseases and conditions related to aging processes or degenerative changes. In addition, it could be used to replace cells and tissues in injured organs. Furthermore, it may provide a novel approach to congenital anomalies and genetic disorders where current therapeutic options are limited However, many crucial questions need answers to ensure a safe, effective and successful solution in the field of cell therapy. In Thailand, innovative knowledge and expertise in stem cell biology and technology are required as the key elements to make cell therapy a "real" hope. PMID- 16696406 TI - Health research, fair benefits and access to medicines. AB - Access to medicines within the context of health research is viewed as a negotiation for 'fair benefits" where it is judged by the host country that access to medicines is the most important and desirable benefit. Research ethics committees in the host country, both local and central, are set to be key players in that determination. However, access to medicines, or "reasonable availability" of medicines in research ethics guidelines, may be difficult to achieve in practice. In extreme cases, the problem of access to medicines may need to be viewed as a global problem, beyond the negotiations within the fair benefits framework. PMID- 16696407 TI - Mental health of residents during Obstetrics and Gynecology training in Thailand. AB - OBJECTIVE: To evaluate mental health of residents during Obstetrics and Gynecology training and identify the factors that affect mental health problems. MATERIAL AND METHOD: During January-April 2004, one hundred and sixty Obstetrics and Gynecology residents (62% of 259) had completed a self-administered questionnaire composed of demographic data, workload, self-report of life stressors, and the Thai Mental Health Questionnaire (TMHQ-70). RESULTS: The prevalence of mental health problems was 29% (46/160). Somatization and social function were found in the first and second rank (18.1% and 11.9%), respectively. A resident who was younger than 25 years old, was married, cared for more than 20 patients per 8 hours in the labor room, performed more than 10 academic activities per year, or attended more than 5 examinations per year was at risk to develop social function problems. CONCLUSION: About one-third of residents training in Obstetrics and Gynecology have faced mental health problems- somatization, social function, depression and anxiety. The significant risk factors associated with social function problems were younger age, marriage and excessive workload PMID- 16696408 TI - Survival rate of recurrent cervical cancer patients. AB - OBJECTIVES: To determine the survival rate of recurrent cervical cancer patients and factors influencing survival. MATERIAL AND METHOD: The subjects were identified from the Gynecologic Oncology Unit tumor registry record The pathological, clinical data including the follow-up information of recurrent cervical cancer patients who were treated in the Gynecologic Oncology Unit, Bangkok Metropolitan Administration Medical College and Vajira Hospital between 1992 and 2003 were retrospective reviewed. RESULTS: During the study period, 144 recurrent cervical cancer patients were identified. Mean age of the patients was 52 years. The median time from complete primary treatment to disease recurrence was 14.8 months. Seventy-two patients (50%) had previous stage III disease. The most common histopalogy was squamous cell carcinoma (72.9%). Approximately half of the recurrences were local (73 patients or 50.7%) and distal recurrences were encountered in 71 patients or 49.3%. Overall 109 patients received treatmentfor their recurrences, i.e. radiation alone (55 patients, 38.2%), chemotherapy (31 patients, 21.5%), chemotherapy and radiation (18 patients, 12.5%), surgery (5 patients, 3.5%), and 35 patients (24.3%) received only supportive treatment. Two year survival rate of the group was 18.5%. Median survival was 8 months (95%CI, 7 10 months). The patients with only local recurrence had a 2-year survival rate of 22.2% compared to 14.6% in those with distant recurrence. (p = 0.245). Two-year survival rate of those who received any kind of treatment was 22.4% compared to 4.0% in those who received only supportive treatment (p = 0.014 and 0.017 in univariable and multivariable analysis respectively). CONCLUSION: Survival rate of recurrent cervical cancer was low, especially in those who received only supportive treatment. PMID- 16696409 TI - Prevalence and associated factors of discordant twins in Siriraj Hospital. AB - OBJECTIVES: To determine the prevalence of discordant twins in Siriraj Hospital and to evaluate associated factors. DESIGN: Cross-sectional study. SETTING: Department of Obstetrics and Gynecology, Faculty of Medicine Siriraj Hospital, Mahidol University. SUBJECTS: A total of 150 women with twin pregnancy, > or = 28 weeks of gestation, who had deliveries of both live twins at Siriraj Hospital from 2003 to 2004. METHOD: A review of medical records was conducted. Discordance was defined as > or = 20% difference in birth weight. Prevalence of discordance was calculated and associated risk factors were evaluated. RESULTS: Discordance was found in 35 cases; therefore, the prevalence was 23.3%. No significant association was found between discordance and various factors, including maternal age, maternal complications, parity, pregnant by assisted reproduction, gestational age at first diagnosis, chorionicity. However, discordant twins delivered at earlier gestational age compared to concordant twins (34.9 +/- 3.2 and 36.2 +/- 2.4 weeks, p = 0.037). Infants of discordant pairs were more likely to be admitted to the NICU than those of concordant pairs (17.1% and 3.9%, p < 0.001, both larger and smaller infants). Other neonatal morbidities were not significantly different. CONCLUSION: Discordance twin was found in 23.3% of the cases. No significant associated risk factor was found. Infants of discordant pairs were more likely to be admitted to the NICU than those of concordant pairs. PMID- 16696410 TI - Prevalence of SIL and SCCA in human immunodeficiency virus-seropositive women at anonymous clinic in Chonburi Hospital. AB - The series of 231 HIV-infected women were recruited in Chonburi Hospital, Thailand. Demographic, gynecologic factors were interviewed Pap smear was performed and classified based on the Bethasda system (1991) by a cytotechnologist. All abnormal Pap smear slides were reviewed by a cytopathologist. The prevalence of LSIL, HSIL and SCCA were 2.2%, 8.3% and 2.2%, respectively. There was statistically significant association between the duration of HIV infection and occurrence of SIL and SCCA. (p = 0.007) CONCLUSION: The present study showed a high prevalence of SIL and SCCA in-HIV-infected women at Chonburi Hospital. PMID- 16696411 TI - The impact of overactive bladder, stress and mixed urinary incontinence on quality of life in Thai postmenopausal women. AB - OBJECTIVE: To assess the quality of life (QOL) of Thai postmenopausal women with overactive bladder (OAB), stress urinary incontinence (SUI) and mixed urinary incontinence (MUI). MATERIAL AND METHOD: A total of 420 Thai postmenopausal women (120 had SUI, 60 MUI, 120 OAB and 120 controls) were recruited QOL was assessed by the Thai version short form 36 (SF-36). RESULTS: There was no significant difference in the patients's characteristics. All domains of SF-36 in women with OAB, SUI and MUI were statistically significantly lower than the control group. OAB and MUI had lower SF-36 scores than SUI in all domains except the role of physical health. When compared between OAB and MUI, the women with MUI had lower SF-36 scores than OAB in all domains (except the physical functioning and general health). CONCLUSION: OAB, SUI and MUI had more significant impaired QOL than control. The author found a greater impact on QUL in OAB and MUI in Thai postmenopausal women. PMID- 16696412 TI - Correlation between colposcopically directed biopsy and large loop excision of the transformation zone and influence of age on the outcome. AB - OBJECTIVES: To assess the accuracy and correlation between Colposcopically Directed Biopsy (CDB) and Large Loop Excision of the Transformation Zone (LLETZ) and the influence of age. MATERIAL AND METHOD: A comparative analysis was conducted from the 352 women referred to Rajavithi Hospital from January 1998 to December 2003, to compare between CDB and LLETZ. Correlation was assessed by the percentage of concordance (accuracy), Spearman's rank correlation coefficient (r) and Kappa statistics (K). A subgroup analysis was performed in women aged > or = 50 years to evaluate the influence of age on the correlation. RESULTS: A 66.2% concordance was found between CDB and LLETZ. The correlation and agreement between the two procedures were low (r = 0.32, p < 0.0001; K = 0.24, p < 0.0001). In women aged > or = 50 years, the accuracy was 60.7% and stronger correlation and agreement were noted (r = 0.50, p < 0.0001; K = 0.31, p = 0.001). CONCLUSION: CDB is an inadequate diagnosis tool for Squamous Intraepithelial Lesion (SIL) in woman below 50 years of age. In women aged 50 years or more with satisfactory colposcopy, the accuracy and correlation between two procedures are not compromised. PMID- 16696413 TI - Cryodamage on sperm chromatin according to different freezing methods, assessed by AO test. AB - OBJECTIVE: To evaluate cryodamage effects on human sperm chromatin, motility and cryosurvival rate after freeze-thawing, compared between liquid nitrogen vapour and computerized program freezer, assessed by acridine orange staining method (AO test). MATERIAL AND METHOD: Fifty semen samples were used. After semen analysis, each semen sample was mixed with cryoprotective media and divided into 2 straws. The first straw was frozen with liquid nitrogen vapour and the second with computerized program freezer. After 1 month of cryostorage, semen samples were thawed. Sperm chromatin integrity, motility, morphology, vitality and sperm cryosurvival rate were determined. RESULTS: DNA damage was significantly greater (p < 0.001) following freezing with liquid nitrogen vapour than with computerized program freezer. Furthermore, the computerized program freezing method significantly provided superior post-thaw sperm motility, vitality and cryosurvival rate, compared with the liquid nitrogen vapour freezing method. CONCLUSION: Computerized program freezing is recommended as a first choice method for routine cryostorage. PMID- 16696414 TI - Oral ibuprofen prophylaxis for symptomatic patent ductus arteriosus of prematurity. AB - BACKGROUND: The oral suspension form of ibuprofen has been shown to have the same efficacy and safety as indomethacin in the treatment of symptomatic PDA, however its role is still questionable in the prophylaxis of symptomatic PDA. OBJECTIVES: 1. To assess the efficacy and safety of the drug in the prevention of symptomatic PDA in premature infants. 2. To study its pharmacokinetics-pharmacodynamics relationship. MATERIAL AND METHOD: A randomized, single-blinded, controlled study was performed on premature neonates with a gestational age between 28-32 weeks, birthweight < or = 1500 grams at the neonatal unit, Queen Sirikit National Institute of Child Health from July 2003 to April 2004. Three doses of ibuprofen suspension or placebo were given 24 hours apart. Clinical evaluation was performed daily until the 28th day of life. Echocardiogram was performed prior to the drug administration, on the 3rd and 7th day of life. RESULTS: There were 22 and 20 cases in the ibuprofen and control group respectively. The epidemiologic data between the groups before enrollment showed no significant differences. Prevalence of symptomatic PDA was lower in the ibuprofen than in the control group without any significant side effects (0/22 vs 5/20, p = 0.015 on day 3 and 0/22 vs 6/20, p = 0.006 on day 7). Comparing with the pharmacokinetic study in older children and adult, the present study revealed nearly the same Cmax but longer Tmax and T1/2 in premature neonates. CONCLUSION: Oral ibuprofen suspension could reduce the prevalence of symptomatic PDA without any significant side effects. PMID- 16696415 TI - Risk factors for hypoxic-ischemic encephalopathy in asphyxiated newborn infants. AB - OBJECTIVE: To determine the risk factors for hypoxic-ischemic encephalopathy (HIE) in asphyxiated newborn infants. MATERIAL AND METHOD: A retrospective study of 17,706 newborns, who were admitted to the Neonatal Unit of King Chulalongkorn Memorial Hospital, from July 1999 till the end of December 2000. 84 infants with perinatal asphyxia were enrolled in the present study. All of the possible risk factors that might have contributed to asphyxia were identified and recorded HIE was diagnosed based on the Modified Sarnat-Sarnat Score for the diagnosis of neonatal encephalopathy. The clinical data of the HIE group were compared with those of the HIE negative group. The categorical data were analyzed for statistical significance (p < 0.05) by Chi-square test or Fisher exact test, or Student t-test. The odds ratio and 95% CI were calculated for those with statistical significance. Stepwise multiple logistic regression analysis used to determine the independent factors that may predispose an infant to HIE. RESULTS: Inappropriate antenatal care (OR 9.4; 95%CI: 2.6-35.4), post-term gestation (OR 7.4; 95%CI: 1.4-34.8), vacuum extraction (OR 5.4; 95%CI: 1.1-26.8), male (OR 4.8; 95%CI: 1.3-19.1), prolapsed cord (p = 0.01) and 1 and 5-minute Apgar scores, (p < 0.0001) were significant risk factors for HIE. However, by multiple regression analysis, only a 5-minute Apgar score was significantly associated with HIE (p = 0.001). CONCLUSION: Sophisticated or expensive equipment is not necessary for the treatment of HIE patient. HIE depends mainly on adequate and effective supportive strategy. The delivery of high risk pregnancies, under obstetric facilities and with appropriate intervention and with good neonatal resuscitation, may prevent the perinatal asphyxia and thereby minimize the occurring of HIE. PMID- 16696416 TI - Results of surgical repair of primary obstructive megaureter. AB - Surgical treatment of primary obstructive megaureter is generally recommended when ureteral dilatation is likely to contribute to symptoms such as pyelonephritis or calculi. The authors' objective for this study was to present the surgical results performing ureteral reimplantation, combined with modified Lich-Gregoir antireflux procedure (using anchoring stitch) over a 10 year period. The average operative time for ureteral reimplantation was 259.64 minutes. The authors' success rate was 92% and all patients had no postoperative ureteral obstruction from intussusception of reimplanted ureter Follow-up postoperative renal sonography at 22 weeks showed a decrease in hydronephrosis in 92%. The authors concluded that the surgical results were comparable with surgical results of other techniques and an anchoring stitch should be considered to prevent postoperative ureteral obstruction from ureteral intussusception. PMID- 16696417 TI - The outcomes and prognostic factors of patients with soft-tissue sarcoma. AB - The present study evaluates the outcomes and prognostic factors in patients with soft-tissue sarcomas of the extremities, trunk, head and neck region. A retrospective study of 104 patients who underwent treatment was conducted on 48 males and 56 females with a mean age of 44.5 years (range, 10-85 years). Seventy eight patients had high-grade sarcomas and most tumors (89.5 percent) were located at the extremities. One hundred patients were treated by surgery and 51 patients were treated by both surgery and radiation therapy. With the median follow-up time of 24.5 months, local recurrence developed in 26 patients (25 percent) and distant metastasis developed in 29 patients (27.9 percent). The actuarial overall 3-year disease-specific survival rate were 74.2 percent. Multivariate statistical analysis revealed that positive surgical margin and occurrence of distant metastasis were significant predictors for overall survival. Positive surgical margin was the only factor that increased the risk of local recurrence and older age (> or = 60 years) was only the factor that increased the risk of distant metastasis. The results reaffirm the importance of the surgical margin where uncontrolled local disease affects the risk of local failure and disease-specific survival. Occurrence of distant metastasis is associated with older age (> or = 60 years) and decreases disease-specific survival of the patients. PMID- 16696418 TI - Effectiveness of oxygenation under the drape in phacoemulsification with intraocular lens implantation: a randomized clinical trial. AB - OBJECTIVE: To compare the oxygen saturation and comfort of the patients undergoing phacoemulsification with intraocular lens implantation (PE/IOL) between the patients with and without oxygenation under the linen drape. SETTING: Institutional practice. MATERIAL AND METHOD: A randomized, triple-blind, clinical equivalence trial was conducted. Eighty consecutive patients, scheduled for PE/IOL under topical or subconjunctival anesthesia, were randomized The patients in group 1 received 5 liters/minute flow of 100% oxygen via tubes attached to the upper chest walls. Sham oxygenation was given to the patients in group 2. Patients, surgeons and investigators were masked. The oxygen saturation was recorded every 10 minutes. The patients rated the comfort score within 15 minutes after operation. RESULTS: In group 1 (n = 37), mean age was 68.2 +/- 10.5 years and mean operative time was 29.4 +/- 7.2 minutes. In group 2 (n = 43), mean age was 66.9 +/- 10.6 years and mean operative time was 27.2 +/- 6.3 minutes. In both groups, the oxygen saturation ranged from the mean of 97.3 to 100% throughout the present study. The 99% confidence intervals of the mean differences of the oxygen saturation at 10, 20, and 30 minutes after draping were -1.60 to 0. 17%, -1.63 to 0.05%, and -2.37 to 0.74%, respectively. The comfort score was 84.9% in group 1 and 89.8% in group 2 (p = 0.61). CONCLUSION: With or without oxygen flow under the drape with the tip of the oxygen tube 1.5 cm away from the chin during PE/IOL, the patients' oxygen saturation is equivalent. The patients' comfort is not statistically different. Oxygenation under the linen drape seems to be unnecessary in uncomplicated cases. PMID- 16696419 TI - Recurrent laryngeal cancer after surgical treatment. AB - OBJECTIVE: Study the presentation pattern of recurrent laryngeal cancer after surgical treatment. MATERIAL AND METHOD: The patterns of recurrent laryngeal cancer after surgical treatment were reviewed: 17 cases were included in the present study. RESULTS: The most common encountered cancers among the recurrent cases were transglottic (59%), supraglottic (35%), and only rarely glottic. 88% were of an advanced stage. The most common site of recurrence were neck node (53%) followed by distant metastasis (29%) and primary recurrence (24%). The onset of recurrence was within a few months of surgery for nodal and primary recurrence and about 2 years for distant metastasis. CONCLUSION: Factors influencing recurrence and appropriate management are discussed. PMID- 16696420 TI - A survey of patients with neuropathic pain at Siriraj Pain Clinic. AB - OBJECTIVE: To examine the profile and treatment pattern of patients with neuropathic pain attending Siriraj Pain Clinic. MATERIAL AND METHOD: A 2-year retrospective study of the prevalence of neuropathic pain, characteristics of the patients and the use of medical treatment. Records of all the patients that attended Siriraj Pain Clinic from September 1, 2002 until September 30, 2004 were reviewed. RESULTS: One thousand three hundred and thirty patients' records were reviewed. Five hundred and three patients were diagnosed as having neuropathic pain. The prevalence of neuropathic pain at Siriraj Pain Clinic during the survey period was 37.8%. The average age of neuropathic pain patients was 54 years. The most common type of neuropathic pain was peripheral type, nerve compression in particular. The most common concomitant illness was malignant tumor. The majority of patients (71.8%) had one pain location and the most common site was the lower extremity. The main descriptions of neuropathic pain were radiating, electric shock-like, burning, numbing and shooting. Oral medication was the most common method (79%) of pain-relief treatment. Almost all of the patients (93%) had received more than one type of oral medication. The most commonly used medicine was TCA (77.1%), followed by gabapentin (35%), carbamazepine (34%) and tramadol (24.3%). Most of the pain-relief medicines prescribed at this clinic were under the recommended doses for the treatment of neuropathic pain. CONCLUSION: Characteristics and treatment patterns of neuropathic pain at Siriraj Pain Clinic are similar to those seen in other pain clinics elsewhere in the world. The high prevalence of neuropathic pain in the clinic indicates that this type of pain syndrome is increasingly critical to our clinical practice. More educational programs on neuropathic pain and management are needed for Thai healthcare professionals. PMID- 16696421 TI - Seizure presenting to the emergency department, Srinagarind Hospital. AB - The authors would like to evaluate the incidence, clinical manifestation, the appropriateness of treatment, and outcome of seizure at the emergency department (ED). All charts of patients who visited the ED of Srinagarind Hospital from 1 January 2003 to 31 December 2003 were reviewed. The profiles of patients and management at the ED were recorded. There were 33,508 cases who visited the ED with 104 cases (0.31%) presenting with seizure. Four cases (3.9%) were diagnosed as status epilepticus. Generalized tonic-clonic seizure was the most common type. Poor antiepileptic drug (AED) compliance with the low AED level was the main precipitating factor. The normal physical examination and routine laboratory tests were normal in the majority of patients. Phenytoin intravenous loading was the commonest initial treatment even in patients with non-status epilepticus. Fourteen patients (13.5%) were treated with intravenous diazepam even though seizures were discontinued. Sixty patients (57.7%) were discharged after seizure was controlled. The advice in seizure control was recorded in only 11 cases (10.6%). From this review, 12 patients presented at the ED for continuing medication without any seizures. In conclusion, seizure at the ED should be treated more appropriately with both laboratory investigation and drug treatment. Futhermore, patient education should be implemented. PMID- 16696422 TI - Glove allergy and sensitization to natural rubber latex among nursing staff at Srinagarind Hospital, Khon Kaen, Thailand. AB - BACKGROUND: According to studies from different countries, the prevalence of natural rubber latex (NRL) sensitization in healthcare workers ranges from 2.9 to 17%. The incidence and prevalence of sensitization and allergy to NRL in Thailand is limited to two studies. There is no study among the high risk healthcare workers. OBJECTIVES: 1) To estimate the prevalence of natural rubber latex (NRL) glove allergy and NRL sensitization among nurses; 2) To describe its clinical symptoms. MATERIAL AND METHOD: Included in the present study were 412 nursing and medical record staff. A self-administered questionnaire was used to collect personal biodata and individual allergy histories to NRL products. Skin prick tests (SPTs) with the commercial NRL allergens; Stallergenes, S.A, Fresnes, France, and common environmental allergens, were performed. RESULTS: The questionnaire response rate was 88% (412/470), 93% females. The response rate of SPT was 72% (295/412) (95%CI 67.2, 76.0). The prevalence of NRL glove allergic symptoms and NRL sensitization was 24% (95%CI 19.9, 28.1) and 2% (95%CI 0.4, 3.6), respectively. The mostfrequently reported symptoms among the positive SPT to NRL was angioedema. Five of the six NRL sensitised subjects had had high exposure to NRL. CONCLUSION: NRL sensitization among Thai nursing staff is less prevalent than in healthcare workers in Europe. PMID- 16696423 TI - Malignant peripheral nerve sheath tumor of breast in patient without Von Recklinghausen's neurofibromatosis: a case report. AB - Malignant peripheral nerve sheath tumor (MPNST) of the breast without Von Recklinghausen's neurofibromatosis (VRN) is extremely rare. The authors report a 19 year-old woman who presented with a 12 month history of a painless mass of the left breast. Tissue biopsy was performed. The histologic diagnosis was made with immunohistochemical study in which the tumor showed positivity of vimentin, S-100 protein, neuron-specific protein(NSE), neurofilament protein(NF) and glial fibrillary acidic protein(GFAP). The patient was referred for radiation therapy after simple mastectomy. PMID- 16696424 TI - Cortical blindness after motorcycle accident: a case report. AB - A 47 year-old Thai man presented with acute bilateral blindness after suffering head trauma in a motorcycle accident. No neurological deficit was found. The visual acuity was light perception with poor light projection on both eyes. Other ocular examinations were normal except ecchymosis on both lower eyelids and mild subconjunctival hemorrhage on the left eye. Emergency computerized tomography showed normal appearance of optic nerves, optic canals and multiple contusional intracerebral hemorrhages at both occipital lobes. This case demonstrated a rare condition of permanent cortical blindness without other neurological deficits resulting from a head trauma. PMID- 16696425 TI - Hepatic artery embolization to control liver hemorrhages by interventional radiologists: experiences from Khon Kaen University. AB - Between 2001 and 2005, nine patients with liver hemorrhage underwent non-surgical embolotherapy. There were six males and three females, ranging in age from 9 to 58 years. All patients were symptomatic with hypotension and decreasing hematocrit. Causes of liver hemorrhage included liver laceration following blunt abdominal injury (2 patients), pseudoaneurysms of hepatic artery following blunt abdominal trauma (4 patients), post-traumatic hepatic artery and portal vein pseudoaneurysm (1 patient), hemobilia following liver resection for cholangiocarcinoma (1 patient) and hepatic artery aneurysm with fistula to the common bile duct (1 patient). All patients had successful embolization of artery to control liver hemorrhage. No acute complication occurred. In conclusion, embolotherapy is an effective treatment with a low complication rate to stop liver hemorrhage, especially from an artery. PMID- 16696426 TI - State of the art: deep brain stimulation in Parkinson's disease. AB - Surgical therapy for Parkinson's disease has a long history beginning in the 1930s with empirical exploration of different brain targets, such as resection of the primary motor cortex or extirpation of the caudate. Recently, there has been a renaissance of functional neurosurgery for the treatment of advanced Parkinson's disease, particularly deep brain stimulation (DBS). To date, DBS of the globus pallidus interna and subthalamic nucleus has been reported to relieve motor symptoms and levodopa-induced dyskinesia in patients with advanced Parkinson's disease. DBS also has different advantages over pallidotomy and subthalamotomy, including reversibility, decreased risk of reoperation and decreased morbidity. In addition to well-experienced neurologists and neurosurgeons, a multidisciplinary team approach is fundamental and critical to ensure success in the DBS procedure in individual patients. With the advances in neuroimaging, neurophysiology and localization techniques, it is increasingly likely that there will be more surgical targets in the future that can also improve cardinal features of Parkinson's disease, or even nonmotor manifestations of this condition. PMID- 16696427 TI - Physician's weak points at court. PMID- 16696428 TI - Myxozoan genera: definition and notes on taxonomy, life-cycle terminology and pathogenic species. AB - A list of myxozoan genera is presented in the current taxonomical scheme. These genera are defined; their type species and most important pathogens along with their hosts are listed. Simultaneously, definitions of actinospore stages representing sexual stages of the myxosporean life cycle are given; altogether, 17 actinospore collective groups with 180 types have been described. Life cycles of the two classes of the phylum Myxozoa, Malacosporea and Myxosporea, are briefly outlined with specification of the appropriate terms. Up to now, 4 malacosporean and 2,180 myxosporean species assigned to a total of 62 genera, have been established. The surviving classification of myxosporeans, based on spore morphology, is discussed in the context of the still fragmentary data resulting from SSU rDNA sequence analyses. The main task for the future is a rigorous, detailed morphological description combined with molecular techniques in establishment of new species and in revision of the existing ones. Establishment of a classification acceptable from morphological, biological and phylogenetical viewpoints is necessary. PMID- 16696429 TI - Occurrence of the microsporidian parasite Nucleospora salmonis in four species of salmonids from the Massif Central of France. AB - Nucleospora salmonis (Hedrick, Groff et Baxa, 1991), an intranuclear microsporidian parasite of marine and freshwater fish, causes diseases mainly in salmonid species. Losses have been reported in stocks of salmonid fish reared in the region of Auvergne (France). The cause of chronic mortalities in the local host species raised in aquaculture and destined for supplementation of the river system Loire-Allier was examined. The presence of N. salmonis was confirmed by PCR and histology in Salmo salar L. previously and in newly investigated salmonid species, Salmo salar, Salmo trutta fario L., Thymallus thymallus (L.) and Salvelinus alpinus (L.), present in European streams. The infection by N. salmonis was consistent in all cases with characteristic symptoms of the disease in deceased or moribund fish. The small subunit ribosomal DNA from N. salmonis was partially sequenced and compared to previously characterised N. salmonis isolates. As a result, a genotype, or clonal entity, was attributed to N. salmonis among Atlantic salmon found along the Northern Atlantic coastal lines and other salmonid species co-inhabiting or co-cultivated in the Auvergne region. PMID- 16696430 TI - Morphological, developmental and ecological evidence for a progenetic life cycle in Neochasmus (Digenea). AB - Seven species of fishes, Catostomus commersonii (Lacepede), Etheostoma nigrum Rafinesque, Micropterus dolomieu Lacepede, Notemigonus crysoleucas (Mitchill), Notropis hudsonius (Clinton), Perca flavescens (Mitchill), and Percina caprodes (Rafinesque) from the St. Lawrence River, Quebec, Canada, were found infected with progenetic specimens of Neochasmus spp. in the orbits and/or the body musculature. Worms displayed varying degrees of maturation. Eggs occupied the entirety of the worm in late stages of development and persisted as distinct clusters in situ after worm death. Populations of parasites were studied monthly in E. nigrum from one site between May and October in order to follow parasite recruitment, development and maturation. Recruitment of parasites was observed in young-of-the-year fish primarily in July and continued through October. Worms matured rapidly, displaying egg production within a month. Later developmental stages, in which eggs occupied most of the worm, and clusters of eggs became abundant by September. Infections in overwintered fish collected in May consisted mainly of worms in early stages of egg production and of clusters of eggs. When hatched artificially, eggs from the clusters released viable miracidia, indicating that they survive beyond the lifespan of the adult worm. It is suggested that progenesis is a fixed characteristic of the life cycle of these species, that egg dispersal requires the death of the host and that it is facilitated by predation. All prior records of Neochasmus spp. are examined, leading us to conclude that the role of the putative definitive host (primarily basses) has been reduced to that of a dispersal agent. Current hypotheses concerning the evolution and maintenance of progenesis are considered, but it is concluded that they do not apply to this host-parasite system. PMID- 16696431 TI - Plectanocotyle major sp. n. (Monogenea: Plectanocotylidae), a gill parasite of Chelidonichthys obscurus (Teleostei: Triglidae) from the Mediterranean Sea. AB - Plectanocotyle major sp. n. (Monogenea: Polyopisthocotylea: Plectanocotylidae) is described from Chelidonichthys obscurus (Bloch et Schneider, 1801) collected from the Mediterranean coasts (Tunisia and France). The new species is compared to Plectanocotyle gurnardi collected on Chelidonichthys gurnardus and the only other valid species in the genus Plectanocotyle. The new species differs from P. gurnardi by the clamp size (137 microm vs 64 microm for clamp length and 110 microm vs 60 microm for clamp width, respectively) and the length of the male copulatory organ (107 microm vs 144 microm, respectively). Species of the Plectanocotylidae seem to be characterized by an oioxenic specificity. PMID- 16696432 TI - The handling and fate of spermatophores in Neoentobdella diadema and N. apiocolpos (Monogenea: Capsalidae: Entobdellinae). AB - The elongated encased spermatophores of the capsalid (entobdelline) monogeneans Neoentobdella diadema (Monticelli, 1902) Kearn et Whittington, 2005 and N. apiocolpos (Euzet et Maillard, 1967) Kearn et Whittington, 2005 have been found attached by their proximal ends to the region of the vaginal opening, with the bulk of the spermatophore projecting from the vagina and therefore lying outside the body. In spite of previous reports, no spermatophores were found projecting from the common genital opening and if spermatophore exchange is as rapid as it is in the related entobdelline Entobdella soleae, then the chances of finding a spermatophore in this location are small. In N. diadema and N. apiocolpos it is likely that sperm enters the vagina through the open proximal end of an attached spermatophore, after which the empty spermatophore case is probably discarded. There is no evidence for a previous proposal that the whole spermatophore is engulfed by the vagina followed by digestion of the case to release the sperm. Three specimens of N. diadema were found each with two spermatophore cases projecting from the vagina and a specimen of N. apiocolpos carried three cases. Assuming that each parasite is able to donate or receive only one spermatophore at each mating, then the presence of one spermatophore does not prevent a further mating and acceptance of a fresh spermatophore. In spite of differences between the spermatophores of E. soleae and N. diadema/N. apiocolpos, the events of spermatophore exchange may be similar. PMID- 16696433 TI - Two new species of Philometra (Nematoda: Philometridae) parasitic in the perciform fish Cynoscion nebulosus (Sciaenidae) in the estuaries of South Carolina, USA. AB - Two new nematode species of Philometra Costa, 1845, P. carolinensis sp. n. and P. cynoscionis sp. n., are described from the spotted seatrout, Cynoscion nebulosus (Cuvier) (Sciaenidae, Perciformes) in estuaries on the Atlantic coast of South Carolina, USA. Philometra carolinensis (males and gravid and subgravid females) parasitic in the host's ovary is most similar to P. lateolabracis (Yamaguti, 1935) in female morphology, but distinctly differs from it in possessing conspicuously short spicules (81-87 microm long) with heavily sclerotized distal parts, the gubernaculum bearing a reflected dorsal barb, and in the length ratio of the gubernaculum and spicules (1:1.23-1.42) in the male. Philometra cynoscionis (only gravid females) found subcutaneously in the host's tissue (lateral to the ascending process of the premaxilla and also posteromedial to the length of each maxilla) is characterized mainly by the presence of cephalic papillae arranged in two circles (8 large papillae in outer circle and 6 small papillae in inner circle), the absence of caudal projections, the body size (length of gravid female 20-33 mm) and the location in the host. PMID- 16696434 TI - Sub-tegumental ventral fibres surrounding the vaginal opening of Entobdella soleae (van Beneden et Hesse, 1864) Johnston, 1929 (Platyhelminthes: Monogenea: Capsalidae). AB - The distribution of F-actin in the monogenean Entobdella soleae (van Beneden et Hesse, 1864) Johnston, 1929 (Platyhelminthes: Capsalidae) was revealed by staining paraformaldehyde-fixed specimens with FITC-labelled phalloidin. On the ventral surface on the left side of the body, just posterior to the pharynx, a concentrically arranged array of fluorescent fibres was observed, following a circular path around the tiny ventral opening of the vagina. It is assumed that this asymmetrically placed array of actin fibres is contractile and the possible role of these fibres in the assimilation of sperm from an attached spermatophore into the vagina is discussed. PMID- 16696435 TI - Ultrastructure of the secondary osmoregulatory canals in the scolex and neck region of Silurotaenia siluri (Batsch, 1786) (Cestoda: Proteocephalidae). AB - The secondary osmoregulatory canals in the scolex and neck region of Silurotaenia siluri, a parasite of the catfish Silurus glanis (L.), terminate below the tegument basal plasma membrane. The basal plasma membrane of the osmoregulatory canal syncytium is in tight contact with the tegument basal plasma membrane. PMID- 16696436 TI - Protein biotoxins of military significance. AB - There is a spectrum of several threat agents, ranging from nerve agents and mustard agents to natural substances, such as biotoxins and new, synthetic, bioactive molecules produced by the chemical industry, to the classical biological warfare agents. The new, emerging threat agents are biotoxins produced by animals, plants, fungi, and bacteria. Many types of organisms produce substances that are toxic to humans. Examples of such biotoxins are botulinum toxin, tetanus toxin, and ricin. Several bioactive molecules produced by the pharmaceutical industry can be even more toxic than are the classical chemical warfare agents. Such new agents, like the biotoxins and bioregulators, often are called mid-spectrum agents. The threat to humans from agents developed by modern chemical synthesis and by genetic engineering also must be considered, since such agents may be more toxic or more effective in causing death or incapacitation than classical warfare agents. By developing effective medical protection and treatment against the most likely chemical and mid-spectrum threat agents, the effects of such agents in a war scenario or following a terrorist attack can be reduced. Toxin-mediated diseases have made humans ill for millennia. Unfortunately, the use of biological agents as weapons of terror has now been realized, and separating naturally occurring disease from bioterroristic events has become an important public health goal. The key to timely identification of such attacks relies on education of primary care physicians, first responders, and public health officials. PMID- 16696437 TI - Apoptotic machinery: the Bcl-2 family proteins in the role of inspectors and superintendents. AB - Programmed cell death, apoptosis, plays an integral role in a variety of biological events, e.g. morphogenesis, removal of unwanted or harmful cells, tissue homeostasis etc. Members of the Bcl-2 family have been described as the key players in the regulation of the apoptotic process. This family consists of proteins that prevent apoptosis (Bcl-2-like) and two structurally distinct subgroups (Bax-like and BH3-only) that on the contrary promote cell death. Majority of their response is concentrated to the mitochondrial level. In this paper, besides reviewing some new information in this field we focused on how they interact among each other and on the way they sense and influence the death signals from the environment. Here, we compare Bcl-2 family to inspectors and superintendents since they supervise the manufacturing process of cell death and they determine whether the cell will die or it will resist and survive. PMID- 16696438 TI - Aesthetics in implantology. AB - Aesthetics in implantology is a topic currently attracting lot of attention from dental implantologists all over the world. It includes both white and red aesthetics with much greater importance been given to the red aesthetics. In this article, both hard tissue and soft tissue procedures and techniques, used in the development of implant recipient sites will be presented and discussed, briefly outlining the treatment planning and also various dental materials used in enhancing aesthetics in implant restorative procedures. PMID- 16696439 TI - Polyunsaturated fatty acids, phytosterols and cholesterol metabolism in the Mediterranean diet. AB - The objective of this work was to review nutritional components of the Mediterranean diet. The Mediterranean diet is not a specific diet plan or diet program but a collection of eating habits that are traditionally followed by the people of the Mediterranean region. There are at least 16 countries bordering the Mediterranean Sea and food habits vary between these countries according to culture, ethnic background and religion. The Mediterranean diet, containing olive oil, fish, fruits and vegetable is associated with a low rate of cardivascular and cancer diseases. This diet is rich in phytosterols, squalene, dietary fibre, antioxidants, phenolic substances and polyunsaturated acids. PMID- 16696440 TI - Mesenchymal stem cells isolated from the human bone marrow: cultivation, phenotypic analysis and changes in proliferation kinetics. AB - Mesenchymal Stem Cells (MSCs) are rare elements living in various organs (e.g., bone marrow), able to differentiate into specialized tissues, such as bone, cartilage, tendon, and myocardium. Since the first description of MSCs by Fridenshtein, several investigators have shown that these cells can also differentiate into chondrocytes, adipocytes, and, at least, in rodents into skeletal myoblasts. Later on, more primitive progenitor cells were characterized, able to give rise not only to limb-bud mesoderm, but also to cells of visceral mesoderm. Those cells were named mesodermal progenitor cells (MPCs). The aim of our study was to characterize and compare the biological properties and spontaneous differentiation potential of two different cell types (MSCs and MPCs) isolated from the human vertebral body bone marrow. The results of our experiments proved that the MPCs can be expanded beyond Hayflick's limit and differed from MSCs in morphology, biological and phenotypic characteristics. Because of their high proliferative and differentiation potential, MPCs can become more attractive source of adult stem cells for therapeutic purposes. PMID- 16696441 TI - Glycation and advanced glycation end-products in laboratory experiments in vivo and in vitro. AB - The purpose of our study was to determine the amount of glycated proteins and advanced glycation end products (AGE) in cataractous lens homogenates of patients who underwent phacoemulsification, and to define a simple in vitro protein model of glycoxidation. Analysis of 30 cataractous lenses (15 diabetic and 15 non diabetic) revealed a significant increase in both glycated lens proteins of diabetics compared with the controls (0.15 vs 0.08 nmol/mg protein, P < 0.01) and AGE-linked fluorescence at 440 nm (4.8 vs 2.8 AU/mg protein, P < 0.01). The presence of AGE fluorescence in lenses indicates the role of oxidative stress in cataractogenesis. Fifty-six days incubation of alanine and aspartate aminotransferases, used as model proteins, with 500 mM D-fructose at 25 and 37 degrees C led to a complete inhibition of ALT and AST activities. The fluorescence of both aminotransferases rose according to the chosen incubation temperature: 37 degrees C > 25 degrees C > 4 degrees C. ALT and AST incubated in a medium containing D-fructose are subject to nonenzymatic glycation followed by a consequent formation of AGE products. Our data: i) support the concept of glycation-glycoxidation pathway appearing in diabetic patients; ii) form a base for determination of the efficiency of various antioxidative compounds in vitro. PMID- 16696442 TI - Prevalence, epidemiological aspects and clinical importance of TT virus infection in Slovakia. AB - The aim of the study was to assess the prevalence, clinical impact and importance of different risk factors of transmission of TTV infection in Slovakia using two PCR methods. Sera of 426 adult persons were examined. TTV DNA was identified by PCR using primers from N22 and untranslated region (UTR) respectively. The established prevalence of TTV tested with N22 and UTR primers according to patients groups was: acute hepatitis of unknown etiology 4 resp. 28 of 37, acute hepatitis B 3 resp. 29 of 38, chronic hepatitis B 11 resp. 41 of 44, chronic hepatitis C 10 resp. 93 of 102, hemodialysis patients 13 resp. 72 of 72, health care workers 0 resp. 27 of 33, control group 8 resp. 83 of 100. Using N22 primers, TTV infection occurred more frequently in chronic hepatitis B group compared with health care workers, if UTR primers were used the group of hemodialysis patients differed significantly from both acute hepatitis groups, health care workers and controls (p < 0.05). From possible risk factors hemodialysis and transfusion count showed notable differences. Bilirubin and aminotransferase levels did not differ between TTV positive and negative groups. No pathogenetic role of TT virus in liver injury was confirmed. PMID- 16696443 TI - Analysis of thrombotic complications after catheter ablation. AB - INTRODUCTION: Thromboembolic complications are described in about 1% of the patients undergoing radiofrequency catheter ablation (RFA). The aim of this study was to analyze thrombotic complications after RFA and to determine prothrombotic states in patients with thrombotic complications. METHODS: We analyzed data from 400 patients (212 females) who underwent 453 RFA procedures for supraventricular tachycardias. Transthoracic echocardiography was performed one day before and after RFA in all patients. We evaluated the clinical and laboratory (in patients with thrombotic complications after RFA) risk factors of thromboembolism. RESULTS: We observed thrombotic complication in 7 (1.75%) patients (6 females), thrice flail thrombus in the right atrium, flail thrombus in the inferior vena cava, femoral vein thrombosis with massive pulmonary embolism, femoral vein mural thrombus and upper extremity digital arteries embolization; four of them were asymptomatic. As a prothrombotic state we identified factor V Leiden mutation in one case and the use of oral contraceptives in two cases. Two other patients had a positive history of thromboembolic events. In a subgroup of females the use of oral contraceptives (p = 0.13) or a positive history of thromboembolism (p = 0.21) were not identified as important risk factors. CONCLUSION: Echocardiographic detection of asymptomatic thrombotic complications contributed to the higher percentage of these complications in our study. Although we can identify the risk factor (laboratory or clinical) in a majority of patients with a thromboembolic complication, occurrence of these complications is unpredictable. PMID- 16696444 TI - Course and prognosis of sarcoidosis in a referral setting in Turkey; analysis of 166 patients. AB - BACKGROUND: Clinical manifestations and prognosis of sarcoidosis are heterogenous and the prevalence varies depending on the country, area and race evaluated. Also the organs involved and courses of the disease differ greatly between countries, areas, races and individuals. AIMS: To investigate the sociodemographic characteristics, clinical presentation and symptoms and to determine the severity and prognosis of sarcoidosis in Turkey as a referral center. METHODS: Between January and July 2003 we retrospectively evaluated the outcome of the patients with sarcoidosis whose first clinical visits were between 1965 and 2003 in the multidisciplinary referral setting RS at Cerrahpasa Medical Faculty of the University of Istanbul. Data collected about each patient included sociodemographic characteristics, clinical presentation, symptoms, date of diagnosis, date and age of onset, method and stage of disease at the date of diagnosis and at the date of last evaluation; the mortality and survival rate were calculated. One hundred and sixty six consecutive patients whose first clinical visits were between 1965 and 2003 in the multidisciplinary RS at Cerrahpasa Medical Faculty were enrolled. We contacted every patient in our cohort by telephone calls or home visits. All those contacted were called back to outpatient clinic for a formal evaluation between June and September 2003. A formal physical examination and thorax radiography were performed in patients who came to the hospital. Their radiological stage, signs, symptoms and associated extrapulmonary manifestations were recorded. RESULTS: At the initial presentation, the mean age of diagnosis was 40.3 years. The 31-40 age group is the group with the highest number of patients. Coughing was the most frequent symptom and erythema nodosum was the most frequent sign in both sexes. Thirty eight percent of patients had extrathoracic involvement. The most frequent extrapulmonary site of involvement was skin. The mortality rate was 11.6% (10.8% in females and 13% in males). Comorbidity was 3% (5 females, 1 male). Females, youngs and patients without extrathoracic involvement had higher survival rates. CONCLUSION: Clinical characteristics, course and prognosis of sarcoidosis vary in different studies. The results may vary accordingly to ethnic, geographic, social and economic conditions. PMID- 16696445 TI - The model of D-galactosamine-induced injury of rat hepatocytes in primary culture. AB - D-galactosamine (GalN) is a highly selective hepatotoxin that causes liver damage similar to human viral hepatitis via depletion of uridine nucleotides, which subsequently diminishes synthesis of RNA and proteins. Model of galactosamine hepatotoxicity is frequently used in animal experiments in vitro. The purpose of our study was to establish the model of GalN-induced hepatocyte injury in in vitro conditions using primocultures of rat hepatocytes as an important pre requisite for further experiments in which we would like to study potential hepatoprotective effect of various substances. Rate of hepatocyte injury was evaluated by morphological changes, changes in cell viability, albumin production, mitochondrial membrane potential, activity of mitochondrial dehydrogenases and glutathione content. Marked dose dependent hepatocyte injury was found after 24-hour incubation with GalN. Based on the results we suggest as an optimal model for short-term toxicity test exposure to GalN for 24 hours in dose of 40 mM. PMID- 16696446 TI - Treatment of postparotidectomy fistulae with fibrin glue. AB - Although fistulae resulting from superficial parotidectomy represent a serious problem, there is still little consensus on the optimal treatment. Some suggest management by applying dressings, while others advise surgery. In recent years several other strategies have been proposed. In this manuscript, we present two cases of postparotidectomy fistulae treated by injection with fibrin tissue glue (Tissucol Duo 500, Baxter AG, Vienna, Austria). Furthermore, a review of the literature is presented. In both patients the fistulae completely healed within months without complications. A literature search revealed that this is the first time postparotidectomy fistulae were treated this way. We conclude that postparotidectomy fistulae can be effectively treated with fibrin glue. Moreover, this treatment is simple, safe and has no side effects. PMID- 16696447 TI - Grade-III slipped capital femoral epiphysis with unstability: a report of three cases. AB - We reviewed the cases of three patients with an unstable grade-III slipped capital femoral epiphysis treated between 2001 and 2003. Clinical records and imaging studies were reviewed for patient's history, anatomic features of the slip, definitive treatment and clinical outcome. The duration of the follow-up ranged from twenty-four to fourty-eight months. PMID- 16696448 TI - Acute generalized exanthematous pustulosis induced by olanzapine. AB - The case of a 56 years old man is presented, who developed acute generalized exanthematous pustulosis 5 days after the introduction of olanzapine 10 mg. Multiple 1-mm pustules appeared on the whole body, concentrated especially on her neck and face. Within 2 days, the eruption was increasingly accompanied by erythema and pruritus. No fever, chills, nausea, vomiting, arthralgias or myalgias were recorded. The diagnosis was corroborated by hystopathology. After 7 days of treatment, olanzapine and valproate were stopped. Concomitantly, cetirizine 20 mg p.o. and methylprednisolone 500 mg i.v. were given once. During the following week betamethasone cream was applied, and the pustular eruption resolved completely. PMID- 16696449 TI - Glimpsing the future. PMID- 16696450 TI - Environmental transmission of SARS at Amoy Gardens. AB - Recent investigations into the March 2003 outbreak of SARS in Hong Kong have concluded that environmental factors played an important role in the transmission of the disease. These studies have focused on a particular outbreak event, the rapid spread of SARS throughout Amoy Gardens, a large, private apartment complex. They have demonstrated that, unlike a typical viral outbreak that is spread through person-to-person contact, the SARS virus in this case was spread primarily through the air. High concentrations of viral aerosols in building plumbing were drawn into apartment bathrooms through floor drains. The initial exposures occurred in these bathrooms. The virus-laden air was then transported by prevailing winds to adjacent buildings at Amoy Gardens, where additional exposures occurred. This article reviews the results of the investigations and provides recommendations for maintenance and other measures that building owners can take to help prevent environmental transmission of SARS and other flulike viruses in their buildings. PMID- 16696451 TI - Five-year surveillance of West Nile and eastern equine encephalitis viruses in Southeastern Virginia. AB - To investigate the occurrence of West Nile virus (WNV) and Eastern equine encephalitis virus (EEE) in southeastern Virginia, the Bureau of Laboratories at the Norfolk Department of Public Health (NDPH) analyzed mosquito pools and the sera of sentinel chickens from the southeastern Virginia area each year from 2000 to 2004. Mosquito pool supernatants were screened for the presence of viral RNA by conventional reverse transcription polymerase chain reaction (RT-PCR) and Taqman RT-PCR with the i-Cycler. Mosquito pools were also tested for virus activity by Vero cell culture. The primary enzootic vector of WNV was Culex (Cx.) pipiens and that of EEE was Culiseta (Cs.) melanura. During the five-year surveillance period, the peak minimum infection rates (MIRs) of WNV and EEE in these mosquito species were 2.7 (2002) and 0.9 (2001), respectively. In 2003, the MIRs in Cs. melanura for WNV and EEE were 0.24 and 0.56, respectively; and the MIR for WNV in Cx. pipiens was 0.64. In 2004, Cs. melanura was less active in the WNV transmission cycle (MIR = 0.07) than was Cx. pipiens (MIR = 1.8), and Cs. melanura was the only vector for EEE (MIR = 0.37). The trend was for EEE activity to peak in July; WNV activity peaked in August. Sentinel-chicken sera were tested for IgM antibodies, and peak IgM seroconversions to these arboviruses were recorded in August 2003 for WNV and in July 2003 for EEE. In 2004, the highest IgM seroconversions to EEE occurred later in August. The overall trend of arbovirus activity was greater in 2003 than in 2004. PMID- 16696452 TI - Incorporating problem-based learning exercises into an environmental health curriculum. AB - Environmental health professionals are often confronted with difficult conflicts involving multiple stakeholders. A problem-based-learning approach was used to engage students in a mock conflict resolution exercise involving the heavily publicized dispute over industrial contamination of an aquifer supplying drinking water to neighborhoods in Woburn, Massachusetts. This dispute provided the basis for the bestselling novel A Civil Action, by Jonathan Harr, as well as a major motion picture bearing the same name. Students were assigned to one of three stakeholder groups: the affected families, an industry consortium, or state and federal regulatory authorities. These stakeholder groups were then directed to participate in alternative-dispute-resolution mock negotiations, with the course instructor serving as a neutral facilitator. The "problem" the students were assigned was to reach a consent agreement that was acceptable to all three stakeholder groups. Two groups of undergraduate students (from different semesters) successfully completed all five phases of this endeavor, with nearly unanimous agreement that the approach was more rapid and potentially less expensive than the civil-litigation process portrayed in the book and movie. The alternative-dispute-resolution approach also provided a less adversarial environment, thereby allowing the industry and community groups to work together to forge a more creative, long-term agreement than that which resulted from the actual civil litigation. PMID- 16696453 TI - Recreational water-illness-prevention = healthy swimming. PMID- 16696454 TI - Computer-aided detection, in its present form, is not an effective aid for screening mammography. For the proposition. PMID- 16696455 TI - Computer-aided detection, in its present form, is not an effective aid for screening mammography. Against the proposition. PMID- 16696456 TI - Time delay study of a CT simulator in respiratory gated CT scanning. AB - In respiratory-gated radiotherapy (RGRT), if the time delay in a computed tomography (CT) simulator and that in a linear accelerator (Linac) are different, the simulation and the treatment cannot be synchronized. In this study, we presented a technique to measure the time delay of the AcQSim CT simulator (Philips Medical Systems, Cleveland, OH) using Varian's Real-Time Positioning Management (RPM) system (Varian Medical Systems, Palo Alto, CA). A respiratory gating platform (REF 91150, Standard Imaging, Inc., Middleton, MI) was first set at the position of amplitude maximum (phase 0). Then a ball of 1.3 cm diameter was put on the platform and set at the CT laser. A single axial scan was acquired across the center of the ball without motion. Then the motion was turned on and single axial scans gated at different phases were acquired with a very narrow gating window. The time between the phase giving a good estimate of the ball and phase 0 is the overall delay time. We found that for AcQSim CT, the overall delay for a single axial scan (with 1 s scan time) is 1.75 s. For multiple axial scans, the overall delay is 1.75 s for the first scan and 0.75 s for the subsequent ones. This demonstrated that the CT mechanical startup delay is 1 s. After the first axial scan, the overall delay per scan is less because CT gantry continuously spins and no mechanical delay exists. We call the overall delay without mechanical part the scanning delay, which basically equals half the scan time (0.5 s for 1 s scan time) plus the gating pulse triggering delay (250 ms). The delays were also verified using metal balls of 1.5 mm diameter set at the amplitude minimum (phase 180, initially). We note that it is the scanning delay rather than the triggering delay that should be compensated when doing motion synchronized radiotherapy. The current interface between the RPM system and the AcQSim CT does not compensate for this scanning delay. PMID- 16696457 TI - Monte Carlo study of photon fields from a flattening filter-free clinical accelerator. AB - In conventional clinical linear accelerators, the flattening filter scatters and absorbs a large fraction of primary photons. Increasing the beam-on time, which also increases the out-of-field exposure to patients, compensates for the reduction in photon fluence. In recent years, intensity modulated radiation therapy has been introduced, yielding better dose distributions than conventional three-dimensional conformal therapy. The drawback of this method is the further increase in beam-on time. An accelerator with the flattening filter removed, which would increase photon fluence greatly, could deliver considerably higher dose rates. The objective of the present study is to investigate the dosimetric properties of 6 and 18 MV photon beams from an accelerator without a flattening filter. The dosimetric data were generated using the Monte Carlo programs BEAMnrc and DOSXYZnrc. The accelerator model was based on the Varian Clinac 2100 design. We compared depth doses, dose rates, lateral profiles, doses outside collimation, total and collimator scatter factors for an accelerator with and without a flatteneing filter. The study showed that removing the filter increased the dose rate on the central axis by a factor of 2.31 (6 MV) and 5.45 (18 MV) at a given target current. Because the flattening filter is a major source of head scatter photons, its removal from the beam line could reduce the out-of-field dose. PMID- 16696459 TI - Detection of breast lesion regions in ultrasound images using wavelets and order statistics. AB - Accurate detection and segmentation of suspicious regions within the complex and irregular tissues of the breast, as depicted with ultrasonic B scans, typically require human analysis and decision making. Tissue characterization methods for classifying suspicious regions often depend on identifying and then accurately segmenting these regions. Motivated by an ultimate goal to automate this critical identification and segmentation step for tissue characterization problems, this work examines ultrasonic signal characteristics between various regions of breast tissue broadly classified as normal tissue and breast lesions. This paper introduces a nonparametric model based on order statistics (OS) estimated from multiresolution (MR) decompositions of energy-normalized subregions. Experimental results demonstrate the classification performance of the OS-based features extracted from the tumor and normal tissue regions in multiple scans from 84 patients, which resulted in a total of 204 tumor regions (from 43 malignant and 161 benign) and 816 normal tissue regions. Performance results indicate that OS based features achieved an area under the receiver-operator characteristic curve of 91% in the discrimination between breast lesions and surrounding normal tissues. PMID- 16696460 TI - Determination of the initial beam parameters in Monte Carlo linac simulation. AB - For Monte Carlo linac simulations and patient dose calculations, it is important to accurately determine the phase space parameters of the initial electron beam incident on the target. These parameters, such as mean energy and radial intensity distribution, have traditionally been determined by matching the calculated dose distributions with the measured dose distributions through a trial and error process. This process is very time consuming and requires a lot of Monte Carlo simulation experience and computational resources. In this paper, we propose an easy, efficient, and accurate method for the determination of the initial beam parameters. We hypothesize that (1) for one type of linacs, the geometry and material of major components of the treatment head are the same; the only difference is the phase space parameters of the initial electron beam incident on the target, and (2) most linacs belong to a limited number of linac types. For each type of linacs, Monte Carlo treatment planning system (MC-TPS) vendors simulate the treatment head and calculate the three-dimensional (3D) dose distribution in water phantom for a grid of initial beam energies and radii. The simulation results (phase space files and dose distribution files) are then stored in a data library. When a MC-TPS user tries to model their linac which belongs to the same type, a standard set of measured dose data is submitted and compared with the calculated dose distributions to determine the optimal combination of initial beam energy and radius. We have applied this method to the 6 MV beam of a Varian 21EX linac. The linac was simulated using EGSNRC/BEAM code and the dose in water phantom was calculated using EGSNRC/DOSXYZ. We have also studied issues related to the proposed method. Several common cost functions were tested for comparing measured and calculated dose distributions, including chi2, mean absolute error, dose difference at the penumbra edge point, slope of the dose difference of the lateral profile, and the newly proposed Kappaalpha factor (defined as the fraction of the voxels with absolute dose difference less than alpha%). It was found that the use of the slope of the lateral profile difference or the difference of the penumbra edge points may lead to inaccurate determination of the initial beam parameters. We also found that in general the cost function value is very sensitive to the simulation statistical uncertainty, and there is a tradeoff between uncertainty and specificity. Due to the existence of statistical uncertainty in simulated dose distributions, it is practically impossible to determine the best energy/radius combination; we have to accept a group of energy/radius combinations. We have also investigated the minimum required data set for accurate determination of the initial beam parameters. We found that the percent depth dose curves along or only a lateral profile at certain depth for a large field size is not sufficient and the minimum data set should include several lateral profiles at various depths as well as the central axis percent depth dose curve for a large field size. PMID- 16696458 TI - Analytic IMRT dose calculations utilizing Monte Carlo to predict MLC fluence modulation. AB - A hybrid dose-computation method is designed which accurately accounts for multileaf collimator (MLC)-induced intensity modulation in intensity modulated radiation therapy (IMRT) dose calculations. The method employs Monte Carlo (MC) modeling to determine the fluence modulation caused by the delivery of dynamic or multisegmental (step-and-shoot) MLC fields, and a conventional dose-computation algorithm to estimate the delivered dose to a phantom or a patient. Thus, it determines the IMRT fluence prediction accuracy achievable by analytic methods in the limit that the analytic method includes all details of the MLC leaf transport and scatter. The hybrid method is validated and benchmarked by comparison with in phantom film dose measurements, as well as dose calculations from two in-house, and two commercial treatment planning system analytic fluence estimation methods. All computation methods utilize the same dose algorithm to calculate dose to a phantom, varying only in the estimation of the MLC modulation of the incident photon energy fluence. Gamma analysis, with respect to measured two-dimensional (2D) dose planes, is used to benchmark each algorithm's performance. The analyzed fields include static and dynamic test patterns, as well as fields from ten DMLC IMRT treatment plans (79 fields) and five SMLC treatment plans (29 fields). The test fields (fully closed MLC, picket fence, sliding windows of different size, and leaf-tip profiles) cover the extremes of MLC usage during IMRT, while the patient fields represent realistic clinical conditions. Of the methods tested, the hybrid method most accurately reproduces measurements. For the hybrid method, 79 of 79 DMLC field calculations have gamma < 1 (3%/3 mm) for more than 95% of the points (per field) while for SMLC fields, 27 of 29 pass the same criteria. The analytic energy fluence estimation methods show inferior pass rates, with 76 of 79 DMLC and 24 of 29 SMLC fields having more than 95% of the test points with gamma < or = 1 (3%/3 mm). Paired one-way ANOVA tests of the gamma analysis results found that the hybrid method better predicts measurements in terms of both the fraction of points with gamma < or = 1 and the average gamma for both 2%/2 mm and 3%/3 mm criteria. These results quantify the enhancement in accuracy in IMRT dose calculations when MC is used to model the MLC field modulation. PMID- 16696461 TI - Continuous intensity map optimization (CIMO): a novel approach to leaf sequencing in step and shoot IMRT. AB - A new leaf-sequencing approach has been developed that is designed to reduce the number of required beam segments for step-and-shoot intensity modulated radiation therapy (IMRT). This approach to leaf sequencing is called continuous-intensity map-optimization (CIMO). Using a simulated annealing algorithm, CIMO seeks to minimize differences between the optimized and sequenced intensity maps. Two distinguishing features of the CIMO algorithm are (1) CIMO does not require that each optimized intensity map be clustered into discrete levels and (2) CIMO is not rule-based but rather simultaneously optimizes both the aperture shapes and weights. To test the CIMO algorithm, ten IMRT patient cases were selected (four head-and-neck, two pancreas, two prostate, one brain, and one pelvis). For each case, the optimized intensity maps were extracted from the Pinnacle3 treatment planning system. The CIMO algorithm was applied, and the optimized aperture shapes and weights were loaded back into Pinnacle. A final dose calculation was performed using Pinnacle's convolution/superposition based dose calculation. On average, the CIMO algorithm provided a 54% reduction in the number of beam segments as compared with Pinnacle's leaf sequencer. The plans sequenced using the CIMO algorithm also provided improved target dose uniformity and a reduced discrepancy between the optimized and sequenced intensity maps. For ten clinical intensity maps, comparisons were performed between the CIMO algorithm and the power-of-two reduction algorithm of Xia and Verhey [Med. Phys. 25(8), 1424-1434 (1998)]. When the constraints of a Varian Millennium multileaf collimator were applied, the CIMO algorithm resulted in a 26% reduction in the number of segments. For an Elekta multileaf collimator, the CIMO algorithm resulted in a 67% reduction in the number of segments. An average leaf sequencing time of less than one minute per beam was observed. PMID- 16696462 TI - Reduction of bias and variance for evaluation of computer-aided diagnostic schemes. AB - Computer-aided diagnostic (CAD) schemes have been developed to assist radiologists in detecting various lesions in medical images. In addition to the development, an equally important problem is the reliable evaluation of the performance levels of various CAD schemes. It is good to see that more and more investigators are employing more reliable evaluation methods such as leave-one out and cross validation, instead of less reliable methods such as resubstitution, for assessing their CAD schemes. However, the common applications of leave-one-out and cross-validation evaluation methods do not necessarily imply that the estimated performance levels are accurate and precise. Pitfalls often occur in the use of leave-one-out and cross-validation evaluation methods, and they lead to unreliable estimation of performance levels. In this study, we first identified a number of typical pitfalls for the evaluation of CAD schemes, and conducted a Monte Carlo simulation experiment for each of the pitfalls to demonstrate quantitatively the extent of bias and/or variance caused by the pitfall. Our experimental results indicate that considerable bias and variance may exist in the estimated performance levels of CAD schemes if one employs various flawed leave-one-out and cross-validation evaluation methods. In addition, for promoting and utilizing a high standard for reliable evaluation of CAD schemes, we attempt to make recommendations, whenever possible, for overcoming these pitfalls. We believe that, with the recommended evaluation methods, we can considerably reduce the bias and variance in the estimated performance levels of CAD schemes. PMID- 16696463 TI - Characterization of the phantom material virtual water in high-energy photon and electron beams. AB - The material Virtual Water has been characterized in photon and electron beams. Range-scaling factors and fluence correction factors were obtained, the latter with an uncertainty of around 0.2%. This level of uncertainty means that it may be possible to perform dosimetry in a solid phantom with an accuracy approaching that of measurements in water. Two formulations of Virtual Water were investigated with nominally the same elemental composition but differing densities. For photon beams neither formulation showed exact water equivalence the water/Virtual Water dose ratio varied with the depth of measurement with a difference of over 1% at 10 cm depth. However, by using a density (range) scaling factor very good agreement (<0.2%) between water and Virtual Water at all depths was obtained. In the case of electron beams a range-scaling factor was also required to match the shapes of the depth dose curves in water and Virtual Water. However, there remained a difference in the measured fluence in the two phantoms after this scaling factor had been applied. For measurements around the peak of the depth-dose curve and the reference depth this difference showed some small energy dependence but was in the range 0.1%-0.4%. Perturbation measurements have indicated that small slabs of material upstream of a detector have a small (<0.1% effect) on the chamber reading but material behind the detector can have a larger effect. This has consequences for the design of experiments and in the comparison of measurements and Monte Carlo-derived values. PMID- 16696464 TI - A novel approach to accurate portal dosimetry using CCD-camera based EPIDs. AB - A new method for portal dosimetry using CCD camera-based electronic portal imaging devices (CEPIDs) is demonstrated. Unlike previous approaches, it is not based on a priori assumptions concerning CEPID cross-talk characteristics. In this method, the nonsymmetrical and position-dependent cross-talk is determined by directly imaging a set of cross-talk kernels generated by small fields ("pencil beams") exploiting the high signal-to-noise ratio of a cooled CCD camera. Signal calibration is achieved by imaging two reference fields. Next, portal dose images (PDIs) can be derived from electronic portal dose images (EPIs), in a fast forward-calculating iterative deconvolution. To test the accuracy of these EPI-based PDIs, a comparison is made to PDIs obtained by scanning diode measurements. The method proved accurate to within 0.2+/-0.7% (1 SD), for on-axis symmetrical and asymmetrical fields with different field widths and homogeneous phantom thicknesses, off-axis Alderson thorax fields and a strongly modulated IMRT field. Hence, the proposed method allows for fast, accurate portal dosimetry. In addition, it is demonstrated that the CEPID cross talk signal is not only induced by optical photon reflection and scatter within the CEPID structure, but also by high-energy back-scattered radiation from CEPID elements (mirror and housing) towards the fluorescent screen. PMID- 16696465 TI - Bone-composition imaging using coherent-scatter computed tomography: assessing bone health beyond bone mineral density. AB - Quantitative analysis of bone composition is necessary for the accurate diagnosis and monitoring of metabolic bone diseases. Accurate assessment of the bone mineralization state is the first requirement for a comprehensive analysis. In diagnostic imaging, x-ray coherent scatter depends upon the molecular structure of tissues. Coherent-scatter computed tomography (CSCT) exploits this feature to identify tissue types in composite biological specimens. We have used CSCT to map the distributions of tissues relevant to bone disease (fat, soft tissue, collagen, and mineral) within bone-tissue phantoms and an excised cadaveric bone sample. Using a purpose-built scanner, we have measured hydroxyapatite (bone mineral) concentrations based on coherent-scatter patterns from a series of samples with varying hydroxyapatite content. The measured scatter intensity is proportional to mineral density in true g/cm3. Repeated measurements of the hydroxyapatite concentration in each sample were within, at most, 2% of each other, revealing an excellent precision in determining hydroxyapatite concentration. All measurements were also found to be accurate to within 3% of the known values. Phantoms simulating normal, over-, and under-mineralized bone were created by mixing known masses of pure collagen and hydroxyapatite. An analysis of the composite scatter patterns gave the density of each material. For each composite, the densities were within 2% of the known values. Collagen and hydroxyapatite concentrations were also examined in a bone-mimicking phantom, incorporating other bone constituents (fat, soft tissue). Tomographic maps of the coherent-scatter properties of each specimen were reconstructed, from which material-specific images were generated. Each tissue was clearly distinguished and the collagen-mineral ratio determined from this phantom was also within 2% of the known value. Existing bone analysis techniques cannot determine the collagen mineral ratio in intact specimens. Finally, to demonstrate the in situ potential of this technique, the mineralization state of an excised normal cadaveric radius was examined. The average collagen-mineral ratio of the cortical bone derived from material-specific images of the radius was 0.53+/-0.04, which is in agreement with the expected value of 0.55 for healthy bones. PMID- 16696466 TI - A novel respiratory detection method based on automated analysis of ultrasound diaphragm video. AB - This paper proposes a novel respiratory detection method based on diaphragm motion measurements using a 2D ultrasound unit. The proposed method extracts a respiratory signal from an automated analysis of the internal diaphragm motion during breathing. The respiratory signal may be used for gating. Ultrasound studies of diaphragm breathing motion were performed on four volunteers. The ultrasound video stream was captured and transferred to a personal computer and decomposed into individual image frames. After straightforward image analysis, region of interest selection, and filtering, the mutual information (MI) and correlation coefficients (CCs) between a selected reference frame and all other frames were computed. The resulting MI and CC values were discovered to produce a signal corresponding to the respiratory cycle in both phase and magnitude. We also studied the diaphragm motion of two volunteers during repeated deep inspiration breath holds (DIBH) and found a slight relaxation motion of the diaphragm during the DIBH, suggesting that the residual motion may be important for treatments delivered at this breathing phase. Applying the proposed respiratory detection method to these ultrasound studies, we found that the MI and CC values demonstrate the relaxation behavior, indicatingthat our method may be used to determine the radiation triggering time for a DIBH technique. PMID- 16696467 TI - Implications of unchanged detection criteria with CAD as second reader of mammograms. AB - In this paper we address the use of computer-aided detection (CAD) systems as second readers in mammography. The approach is based on Bayesian decision theory and its implication for the choice of optimal operating points. The choice of a certain operating point along an ROC curve corresponds to a particular tradeoff between false positives and missed cancers. By minimizing a total risk function given this tradeoff, we determine optimal decision thresholds for the radiologist and CAD system when CAD is used as a second reader. We show that under very general circumstances, the performance of the sequential system is improved if the decision threshold of the latent human decision variable is increased compared to what it would have been in the absence of the CAD system. This means that an initial stricter decision criterion should be applied by the radiologist when CAD is used as a second reader than otherwise. First and foremost, the results in this paper should be interpreted qualitatively, but an attempt is made at quantifying the effect by tuning the model to a prospective study evaluating the use of CAD as a second reader. By making some necessary and plausible assumptions, we are able to estimate the effect of the resulting suboptimal operating point. In this study of 12 860 women, we estimate that a 15% reduction in callbacks for masses could have been achieved with only about a 1.5% relative decrease in sensitivity compared to that without using a stricter initial criterion by the radiologist. For microcalcifications the corresponding values are 7% and 0.2%. PMID- 16696468 TI - Light field and crosshair quality assurance test using a simple lens system. AB - We describe here a simple lens system to test the positioning of the field light source and mylar crosshair for radiation therapy linear accelerators. Ideally the light source for the field light and the crosshair should be centered on the axis of rotation of the collimator. The traditional method for testing this coincidence uses the shadow of the crosshair caused by the field light source. The shadow of the crosshair is dependent on the position of both the field light source and mylar crosshair. Geometrically it is possible for the field light source and the mylar crosshair to be off the axis of rotation of the collimator and still cause the shadow of the crosshair to be on the axis of rotation at some distance. Using a lens system the motion of the field light source and crosshair can be observed in sharp focus independently of one another as the collimator is rotated. PMID- 16696469 TI - Scatter rejection in multislit digital mammography. AB - The scatter to primary ratio (SPR) was measured on a scanning multislit full field digital mammography system for different thickness of breast equivalent material and different tube voltages. Scatter within the detector was measured separately and was found to be the major source of scatter in the assembly. Measured total SPRs below 6% are reported for breast range 3-7 cm. The performance of the multislit assembly is compared to other imaging geometries with different scatter rejection schemes by using the scatter detective quantum efficiency. PMID- 16696470 TI - Investigation of gamma knife image registration errors resulting from misalignment between the patient and the imaging axis. AB - The ability of Leksell GammaPlan to perform stereotactic space localizations with image sets where there is misalignment of the patient's head (stereotactic frame and fiducial apparatus) relative to the computed tomography (CT) scanner coordinate system was studied. Misalignment is sometimes necessary for patient comfort. Results equally apply to magnetic resonance imaging. Seven 0.5 mm diameter CT-visible spheres were rigidly mounted to a string tied tightly at each end to diagonally opposite posts attached to a Leksell stereotactic frame. A standard CT fiducial box was applied to the frame in the usual clinical manner. A baseline CT scan (1 mm slice thickness) was obtained with the fiducial box perfectly aligned with the scanner axis. After localization of the image set, the (x,y,z) coordinate of the center of each sphere was recorded. Repeat CT scans with varying fiducial box misalignments with the imaging axis were subsequently obtained. The mean difference between the base line and the respective coordinates in misaligned geometries was approximately 0.2 mm (sigma=0.2 mm), well within the accuracy of the image sets and the delivery of radiosurgery with the Gamma Knife. PMID- 16696471 TI - Constrained segment shapes in direct-aperture optimization for step-and-shoot IMRT. AB - Previous studies have shown that, by optimizing segment shapes and weights directly, without explicitly optimizing fluence profiles, effective IMRT plans can be generated with fewer segments. This study proposes a method of direct aperture optimization with aperture shape constraints, which is designed to provide segmental IMRT plans using a minimum of simple, regular segments. The method uses a cubic function to create smoothly curving multileaf collimator shapes. Constraints on segment dimension and equivalent square are applied, and each segment can be constrained to lie within the previous one, for easy generation of fluence profiles with a single maximum. To simply optimize the segment shapes and reject any shapes which violate the constraints is too inefficient, so an innovative method of feedback optimization is used to ensure in advance that viable aperture shapes are generated. The algorithm is demonstrated using a simple cylindrical phantom consisting of a hemi-annular planning target volume and a central cylindrical organ-at-risk. A simple IMRT rectum case is presented, where segments are used to replace a wedge. More complex cases of prostate and seminal vesicles and prostate and pelvic nodes are also shown. The algorithm produces effective plans in each case with three to five segments per beam. For the simple plans, the constraint that each segment should be contained within the previous one adds additional simplicity to the plan, for a small reduction in plan quality. This study confirms that direct aperture optimization gives efficient solutions to the segmental IMRT inverse problem and provides a method for generating simple apertures. By using such a method, the workload of IMRT verification may be reduced and simplified, as verification of fluence profiles from individual beams may be eliminated. PMID- 16696472 TI - A semianalytic model to extract differential linear scattering coefficients of breast tissue from energy dispersive x-ray diffraction measurements. AB - The goal of this work is to develop a technique to measure the x-ray diffraction signals of breast biopsy specimens. A biomedical x-ray diffraction technology capable of measuring such signals may prove to be of diagnostic use to the medical field. Energy dispersive x-ray diffraction measurements coupled with a semianalytical model were used to extract the differential linear scattering coefficients [mus(x)] of breast tissues on absolute scales. The coefficients describe the probabilities of scatter events occuring per unit length of tissue per unit solid angle of detection. They are a function of the momentum transfer argument, x=sin(theta/2)/X, where theta=scatter angle and lambda=incident wavelength. The technique was validated by using a 3 mm diameter 50 kV polychromatic x-ray beam incident on a 5 mm diameter 5 mm thick sample of water. Water was used because good x-ray diffraction data are available in the literature. The scatter profiles from 6 degrees to 15 degrees in increments of 1 degrees were measured with a 3 mm x 3 mm x 2 mm thick cadmium zinc telluride detector. A 2 mm diameter Pb aperture was placed on top of the detector. The target to detector distance was 29 cm and the duration of each measurement was 10 min. Ensemble averages of the results compare well with the gold standard data of A. H. Narten ["X-ray diffraction data on liquid water in the temperature range 4 degrees C-200 degrees C," ORNL Report No. 4578 (1970)]. An average 7.68% difference for which most of the discrepancies can be attributed to the background noise at low angles was obtained. The preliminary measurements of breast tissue are also encouraging. PMID- 16696473 TI - Potential impact of prostate edema on the dosimetry of permanent seed implants using the new 131Cs (model CS-1) seeds. AB - Our aim in this work was to study the potential dosimetric effect of prostate edema on the accuracy of conventional pre- and post-implant dosimetry for prostate seed implants using the newly introduced 131Cs seed, whose radioactive decay half-life (approximately 9.7 days) is directly comparable to the average edema resolution half-life (approximately 10 days) observed previously by Waterman et al. for 125I implants [Int. J. Radiat. Oncol. Biol. Phys. 41, 1069 1077 (1998)]. A systematic calculation of the relative dosimetry effect of prostate edema on the 131Cs implant was performed by using an analytic solution obtained previously [Int. J. Radiat. Oncol. Biol. Phys. 47, 1405-1419 (2000)]. It was found that conventional preimplant dosimetry always overestimates the true delivered dose as it ignores the temporary increase of the interseed distance caused by edema. The overestimation for 131Cs implants ranged from 1.2% (for a small edema with a magnitude of 10% and a half-life of 2 days) to approximately 45% (for larger degree edema with a magnitude of 100% and a half-life of 25 days). The magnitude of pre- and post-implant dosimetry error for 131Cs implants was found to be similar to that of 103Pd implants for typical edema characteristics (magnitude < 100%, and half-life <25 days); both of which are worse compared to 125I implants. The preimplant dosimetry error for 131Cs implants cannot be compensated effectively without knowing the edema characteristics before the seed implantation. On the other hand, the error resulted from a conventional post-implant dosimetry can be minimized (to within +/-6%) for 131Cs implants if the post-implant dosimetry is performed at 10+/-2 days post seed implantation. This "optimum" post-implant dosimetry time is shorter than those determined previously for the 103Pd and 125I implants at 16+/ 4 days and 6+/-1 weeks, respectively. PMID- 16696474 TI - Method for transforming CT images for attenuation correction in PET/CT imaging. AB - A tube-voltage-dependent scheme is presented for transforming Hounsfield units (HU) measured by different computed tomography (CT) scanners at different x-ray tube voltages (kVp) to 511 keV linear attenuation values for attenuation correction in positron emission tomography (PET) data reconstruction. A Gammex 467 electron density CT phantom was imaged using a Siemens Sensation 16-slice CT, a Siemens Emotion 6-slice CT, a GE Lightspeed 16-slice CT, a Hitachi CXR 4-slice CT, and a Toshiba Aquilion 16-slice CT at kVp ranging from 80 to 140 kVp. All of these CT scanners are also available in combination with a PET scanner as a PET/CT tomograph. HU obtained for various reference tissue substitutes in the phantom were compared with the known linear attenuation values at 511 keV. The transformation, appropriate for lung, soft tissue, and bone, yields the function 9.6 x 10(-5). (HU+ 1000) below a threshold of approximately 50 HU and a (HU+ 1000)+b above the threshold, where a and b are fixed parameters that depend on the kVp setting. The use of the kVp-dependent scaling procedure leads to a significant improvement in reconstructed PET activity levels in phantom measurements, resolving errors of almost 40% otherwise seen for the case of dense bone phantoms at 80 kVp. Results are also presented for patient studies involving multiple CT scans at different kVp settings, which should all lead to the same 511 keV linear attenuation values. A linear fit to values obtained from 140 kVp CT images using the kVp-dependent scaling plotted as a function of the corresponding values obtained from 80 kVp CT images yielded y = 1.003 x -0.001 with an R2 value of 0.999, indicating that the same values are obtained to a high degree of accuracy. PMID- 16696475 TI - Imaging properties of digital magnification radiography. AB - Flat panel detectors exhibit improved signal-to-noise ratio (SNR) and display capabilities compared to film. This improvement necessitates a new evaluation of optimal geometry for conventional projection imaging applications such as digital projection mammography as well as for advanced x-ray imaging applications including cone-beam computed tomography (CT), tomosynthesis, and mammotomography. Such an evaluation was undertaken in this study to examine the effects of x-ray source distribution, inherent detector resolution, magnification, scatter rejection, and noise characteristics including noise aliasing. A model for x-ray image acquisition was used to develop generic results applicable to flat panel detectors with similar x-ray absorption characteristics. The model assumed a Gaussian distribution for the focal spot and a rectangular distribution for a pixel. A generic model for the modulated transfer function (MTF) of indirect flat panel detectors was derived by a nonlinear fit of empirical receptor data to the Burgess model for phosphor MTFs. Noise characteristics were investigated using a generic noise power spectrum (NPS) model for indirect phosphor-based detectors. The detective quantum efficiency (DQE) was then calculated from the MTF and NPS models. The results were examined as a function of focal spot size (0.1, 0.3, and 0.6 mm) and pixel size (50, 100, 150, and 200 microm) for magnification ranges 1 to 3. Mammography, general radiography (also applicable to mammotomography), and chest radiography applications were explored using x-ray energies of 28, 74, and 120 kVp, respectively. Nodule detection was examined using the effective point source scatter model, effective DQE, and the Hotelling SNR2 efficiency. Results indicate that magnification can potentially improve the signal and noise performance of digital images. Results also show that a cross over point occurs in the spatial frequency above and below which the effects of magnification differ indicating that there are task dependent tradeoffs associated with magnification. The cross over point varies depending upon focal spot size, pixel size, x-ray energy, and source-to-image-distance (SID). For mammography, the cross over point occurs for a 0.3 mm focal spot while a 0.6 mm focal spot indicates that magnification does not improve image quality due to focal spot blurring. Thus, the benefit of magnification may be limited. For general radiography (as well as mammotomography), and chest radiography, the cross over point changes with SID. For a system with a 0.3 mm focal spot, 100 microm pixel size, a 2 m SID, and the applicable tissue thickness and scatter components, optimal magnification improved SNR2 by approximately 1.2 times for mammography and 1.5 times for general radiography (and mammotomography). These results indicate that the optimal geometry can improve image quality without changing patient dose or otherwise reduce dose without compromising image quality. PMID- 16696476 TI - An analytical algorithm for skew-slit imaging geometry with nonuniform attenuation correction. AB - The pinhole collimator is currently the collimator of choice in small animal single photon emission computed tomography (SPECT) imaging because it can provide high spatial resolution and reasonable sensitivity when the animal is placed very close to the pinhole. It is well known that if the collimator rotates around the object (e.g., a small animal) in a circular orbit to form a cone-beam imaging geometry with a planar trajectory, the acquired data are not sufficient for an exact artifact-free image reconstruction. In this paper a novel skew-slit collimator is mounted instead of the pinhole collimator in order to significantly reduce the image artifacts caused by the geometry. The skew-slit imaging geometry is a more generalized version of the pinhole imaging geometry. The multiple pinhole geometry can also be extended to the multiple-skew-slit geometry. An analytical algorithm for image reconstruction based on the tilted fan-beam inversion is developed with nonuniform attenuation compensation. Numerical simulation shows that the axial artifacts are evidently suppressed in the skew slit images compared to the pinhole images and the attenuation correction is effective. PMID- 16696477 TI - Two-dimensional ionization chamber arrays for IMRT plan verification. AB - In this paper we describe a concept for dosimetric treatment plan verification using two-dimensional ionization chamber arrays. Two different versions of the 2D ARRAY (PTW-Freiburg, Germany) will be presented, a matrix of 16 x 16 chambers (chamber cross section 8 mm x 8 mm; the distance between chamber centers, 16 mm) and a matrix of 27 x 27 chambers (chamber cross section 5 mm x 5 mm; the distance between chamber centers is 10 mm). The two-dimensional response function of a single chamber is experimentally determined by scanning it with a slit beam. For dosimetric plan verification, the expected two-dimensional distribution of the array signals is calculated via convolution of the planned dose distribution, obtained from the treatment planning system, with the two-dimensional response function of a single chamber. By comparing the measured two-dimensional distribution of the array signals with the expected one, a distribution of deviations is obtained that can be subjected to verification criteria, such as the gamma index criterion. As an example, this verification method is discussed for one sequence of an IMRT plan. The error detection capability is demonstrated in a case study. Both versions of two-dimensional ionization chamber arrays, together with the developed treatment plan verification strategy, have been found to provide a suitable and easy-to-handle quality assurance instrument for IMRT. PMID- 16696478 TI - Generalized five-dimensional dynamic and spectral factor analysis. AB - We have generalized the spectral factor analysis and the factor analysis of dynamic sequences (FADS) in SPECT imaging to a five-dimensional general factor analysis model (5D-GFA), where the five dimensions are the three spatial dimensions, photon energy, and time. The generalized model yields a significant advantage in terms of the ratio of the number of equations to that of unknowns in the factor analysis problem in dynamic SPECT studies. We solved the 5D model using a least-squares approach. In addition to the traditional non-negativity constraints, we constrained the solution using a priori knowledge of both time and energy, assuming that primary factors (spectra) are Gaussian-shaped with full width at half-maximum equal to gamma camera energy resolution. 5D-GFA was validated in a simultaneous pre-/post-synaptic dual isotope dynamic phantom study where 99mTc and 123I activities were used to model early Parkinson disease studies. 5D-GFA was also applied to simultaneous perfusion/dopamine transporter (DAT) dynamic SPECT in rhesus monkeys. In the striatal phantom, 5D-GFA yielded significantly more accurate and precise estimates of both primary 99mTc (bias=6.4 % +/- 4.3 %) and 1231 (-1.7% +/- 6.9%) time activity curves (TAC) compared to conventional FADS (biases = 15.5% +/- 10.6% in 99mTc and 8.3% +/- 12.7% in 123I, p < 0.05). Our technique was also validated in two primate dynamic dual isotope perfusion/DAT transporter studies. Biases of 99mTc-HMPAO and 123I-DAT activity estimates with respect to estimates obtained in the presence of only one radionuclide (sequential imaging) were significantly lower with 5D-GFA (9.4% +/- 4.3% for 99mTc-HMPAO and 8.7% +/-4.1% for 123I-DAT) compared to biases greater than 15% for volumes of interest (VOI) over the reconstructed volumes (p < 0.05). 5D-GFA is a novel and promising approach in dynamic SPECT imaging that can also be used in other modalities. It allows accurate and precise dynamic analysis while compensating for Compton scatter and cross-talk. PMID- 16696479 TI - Effect of edema, relative biological effectiveness, and dose heterogeneity on prostate brachytherapy. AB - Many factors influence response in low-dose-rate (LDR) brachytherapy of prostate cancer. Among them, edema, relative biological effectiveness (RBE), and dose heterogeneity have not been fully modeled previously. In this work, the generalized linear-quadratic (LQ) model, extended to account for the effects of edema, RBE, and dose heterogeneity, was used to assess these factors and their combination effect. Published clinical data have shown that prostate edema after seed implant has a magnitude (ratio of post- to preimplant volume) of 1.3-2.0 and resolves exponentially with a half-life of 4-25 days over the duration of the implant dose delivery. Based on these parameters and a representative dose-volume histogram (DVH), we investigated the influence of edema on the implant dose distribution. The LQ parameters (alpha=0.15 Gy(-1) and alpha/beta=3.1 Gy) determined in earlier studies were used to calculate the equivalent uniform dose in 2 Gy fractions (EUD2) with respect to three effects: edema, RBE, and dose heterogeneity for 125I and 103Pd implants. The EUD2 analysis shows a negative effect of edema and dose heterogeneity on tumor cell killing because the prostate edema degrades the dose coverage to tumor target. For the representative DVH, the V100 (volume covered by 100% of prescription dose) decreases from 93% to 91% and 86%, and the D90 (dose covering 90% of target volume) decrease from 107% to 102% and 94% of prescription dose for 125I and 103Pd implants, respectively. Conversely, the RBE effect of LDR brachytherapy [versus external-beam radiotherapy (EBRT) and high-dose-rate (HDR) brachytherapy] enhances dose effect on tumor cell kill. In order to balance the negative effects of edema and dose heterogeneity, the RBE of prostate brachytherapy was determined to be approximately 1.2-1.4 for 125I and 1.3-1.6 for 103Pd implants. These RBE values are consistent with the RBE data published in the literature. These results may explain why in earlier modeling studies, when the effects of edema, dose heterogeneity, and RBE were all ignored simultaneously, prostate LDR brachytherapy was reported to show an overall similar dose effect as EBRT and HDR brachytherapy, which are independent of edema and RBE effects and have a better dose coverage. PMID- 16696480 TI - Normalized dose data for upper gastrointestinal tract contrast studies performed to infants. AB - The aim of the current study was to (a) provide normalized dose data for the estimation of the radiation dose from upper gastrointestinal tract contrast (UGIC) studies carried out to infants and (b) estimate the average patient dose and risks associated with radiation from UGIC examinations performed in our institution. Organ and effective doses, normalized to entrance skin dose (ESD) and dose area product (DAP) were estimated for UGIC procedures utilizing the Monte Carlo N-particle (MCNP) transport code and two mathematical phantoms, one corresponding to the size of a newborn and one to the size of a 1-year-old child. The validity of the MCNP results was verified by comparison with dose data obtained in physical anthropomorphic phantoms simulating a newborn and a 1-year old infant using thermoluminescence dosimetry (TLD). Data were also collected from 25 consecutive UGIC examinations performed to infants. Study participants were (a) 12 infants aged from 0.5 to 5.9 months (group 1) and (b) 13 infants aged from 6 to 15 months (group 2). For each examination, ESD and dose to comforters were measured using TLD. Patient effective doses were estimated using normalized dose data obtained in the simulation study. The risk for fatal cancer induction was estimated using appropriate coefficients. The results consist of tabulated dose data normalized to ESD or DAP for the estimation of patient dose. Conversion coefficients were estimated for various tube potentials and beam filtration values. The mean total fluoroscopy time was 1.26 and 1.62 min for groups 1 and 2, respectively. The average effective dose was 1.6 mSv for group 1 and 1.9 mSv for group 2. The risk of cancer attributable to the radiation exposure associated with a typical UGIC study was found to be up to 3 per 10 000 infants undergoing an UGIC examination. The mean radiation dose absorbed by the hands of comforters was 47 microGy. In conclusion, estimation of radiation doses associated with UGIC studies performed to infants can be made using the normalized dose data provided in the current study. Radiation dose values associated with UGIC examinations carried out to infants are not low and should be minimized as much as possible. PMID- 16696481 TI - A computer simulation study comparing lesion detection accuracy with digital mammography, breast tomosynthesis, and cone-beam CT breast imaging. AB - Although conventional mammography is currently the best modality to detect early breast cancer, it is limited in that the recorded image represents the superposition of a three-dimensional (3D) object onto a 2D plane. Recently, two promising approaches for 3D volumetric breast imaging have been proposed, breast tomosynthesis (BT) and CT breast imaging (CTBI). To investigate possible improvements in lesion detection accuracy with either breast tomosynthesis or CT breast imaging as compared to digital mammography (DM), a computer simulation study was conducted using simulated lesions embedded into a structured 3D breast model. The computer simulation realistically modeled x-ray transport through a breast model, as well as the signal and noise propagation through a CsI based flat-panel imager. Polyenergetic x-ray spectra of Mo/Mo 28 kVp for digital mammography, Mo/Rh 28 kVp for BT, and W/Ce 50 kVp for CTBI were modeled. For the CTBI simulation, the intensity of the x-ray spectra for each projection view was determined so as to provide a total average glandular dose of 4 mGy, which is approximately equivalent to that given in conventional two-view screening mammography. The same total dose was modeled for both the DM and BT simulations. Irregular lesions were simulated by using a stochastic growth algorithm providing lesions with an effective diameter of 5 mm. Breast tissue was simulated by generating an ensemble of backgrounds with a power law spectrum, with the composition of 50% fibroglandular and 50% adipose tissue. To evaluate lesion detection accuracy, a receiver operating characteristic (ROC) study was performed with five observers reading an ensemble of images for each case. The average area under the ROC curves (Az) was 0.76 for DM, 0.93 for BT, and 0.94 for CTBI. Results indicated that for the same dose, a 5 mm lesion embedded in a structured breast phantom was detected by the two volumetric breast imaging systems, BT and CTBI, with statistically significant higher confidence than with planar digital mammography, while the difference in lesion detection between BT and CTBI was not statistically significant. PMID- 16696482 TI - Segmented crystalline scintillators: empirical and theoretical investigation of a high quantum efficiency EPID based on an initial engineering prototype CsI(TI) detector. AB - Modern-day radiotherapy relies on highly sophisticated forms of image guidance in order to implement increasingly conformal treatment plans and achieve precise dose delivery. One of the most important goals of such image guidance is to delineate the clinical target volume from surrounding normal tissue during patient setup and dose delivery, thereby avoiding dependence on surrogates such as bony landmarks. In order to achieve this goal, it is necessary to integrate highly efficient imaging technology, capable of resolving soft-tissue contrast at very low doses, within the treatment setup. In this paper we report on the development of one such modality, which comprises a nonoptimized, prototype electronic portal imaging device (EPID) based on a 40 mm thick, segmented crystalline CsI(Tl) detector incorporated into an indirect-detection active matrix flat panel imager (AMFPI). The segmented detector consists of a matrix of 160 x 160 optically isolated, crystalline CsI(Tl) elements spaced at 1016 microm pitch. The detector was coupled to an indirect detection-based active matrix array having a pixel pitch of 508 microm, with each detector element registered to 2 x 2 array pixels. The performance of the prototype imager was evaluated under very low-dose radiotherapy conditions and compared to that of a conventional megavoltage AMFPI based on a Lanex Fast-B phosphor screen. Detailed quantitative measurements were performed in order to determine the x-ray sensitivity, modulation transfer function, noise power spectrum, and detective quantum efficiency (DQE). In addition, images of a contrast-detail phantom and an anthropomorphic head phantom were also acquired. The prototype imager exhibited approximately 22 times higher zero-frequency DQE (approximately 22%) compared to that of the conventional AMFPI (approximately 1%). The measured zero-frequency DQE was found to be lower than theoretical upper limits (approximately 27%) calculated from Monte Carlo simulations, which were based solely on the x-ray energy absorbed in the detector-indicating the presence of optical Swank noise. Moreover, due to the nonoptimized nature of this prototype, the spatial resolution was observed to be significantly lower than theoretical expectations. Nevertheless, due to its high quantum efficiency (approximately 55%), the prototype imager exhibited significantly higher DQE than that of the conventional AMFPI across all spatial frequencies. In addition, the frequency-dependent DQE was observed to be relatively invariant with respect to the amount of incident radiation, indicating x-ray quantum limited behavior. Images of the contrast detail phantom and the head phantom obtained using the prototype system exhibit good visualization of relatively large, low-contrast features, and appear significantly less noisy compared to similar images from a conventional AMFPI. Finally, Monte Carlo-based theoretical calculations indicate that, with proper optimization, further, significant improvements in the DQE performance of such imagers could be achieved. It is strongly anticipated that the realization of optimized versions of such very high-DQE EPIDs would enable megavoltage projection imaging at very low doses, and tomographic imaging from a "beam's eye view" at clinically acceptable doses. PMID- 16696483 TI - Improving delivery of segments with small MU in step-and-shoot IMRT. AB - The purpose of this study is to describe four new delivery schemes for intensity modulated radiation therapy (IMRT). In the first two schemes the order in which segments are delivered is varied from fraction to fraction. The last two delivery schemes employ fixed order of segments. The obtained results indicate that the suggested approaches can significantly reduce the so-called "overshoot" and "undershoot" phenomena and the associated discrepancies between planned and delivered monitor units. PMID- 16696484 TI - The US radiation dosimetry standards for 60Co therapy level beams, and the transfer to the AAPM accredited dosimetry calibration laboratories. AB - This work reports the transfer of the primary standard for air kerma from the National Institute of Standards and Technology (NIST) to the secondary laboratories accredited by the American Association of Physics in Medicine (AAPM). This transfer, performed in August of 2003, was motivated by the recent revision of the NIST air-kerma standards for 60Co gamma-ray beams implemented on July 1, 2003. The revision involved a complete recharacterization of the two NIST therapy-level 60Co gamma-ray beam facilities, resulting in new values for the air kerma rates disseminated by the NIST. Some of the experimental aspects of the determination of the new air-kerma rates are briefly summarized here; the theoretical aspects have been described in detail by Seltzer and Bergstrom ["Changes in the U.S. primary standards for the air-kerma from gamma-ray beams," J. Res. Natl. Inst. Stand. Technol. 108, 359-381 (2003)]. The standard was transferred to reference-class chambers submitted by each of the AAPM Accredited Dosimetry Calibration Laboratories (ADCLs). These secondary-standard instruments were then used to characterize the 60Co gamma-ray beams at the ADCLs. The values of the response (calibration coefficient) of the ADCL secondary-standard ionization chambers are reported and compared to values obtained prior to the change in the NIST air-kerma standards announced on July 1, 2003. The relative change is about 1.1% for all of these chambers, and this value agrees well with the expected change in chambers calibrated at the NIST or at any secondary standard laboratory traceable to the new NIST standard. PMID- 16696485 TI - A bench-top megavoltage fan-beam CT using CdWO4-photodiode detectors. I. System description and detector characterization. AB - We describe the components of a bench-top megavoltage computed tomography (MVCT) scanner that uses an 80-element detector array consisting of CdWO4 scintillators coupled to photodiodes. Each CdWO4 crystal is 2.75 x 8 x 10 mm3. The detailed design of the detector array, timing control, and multiplexer are presented. The detectors show a linear response to dose (dose rate was varied by changing the source to detector distance) with a correlation coefficient (R2) nearly unity with the standard deviation of signal at each dose being less than 0.25%. The attenuation of a 6 MV beam by solid water measured by this detector array indicates a small, yet significant spectral hardening that needs to be corrected before image reconstruction. The presampled modulation transfer function is strongly affected by the detector's large pitch and a large improvement can be obtained by reducing the detector pitch. The measured detective quantum efficiency at zero spatial frequency is 18.8% for 6 MV photons which will reduce the dose to the patient in MVCT applications. The detector shows a less than a 2% reduction in response for a dose of 24.5 Gy accumulated in 2 h; however, the lost response is recovered on the following day. A complete recovery can be assumed within the experimental uncertainty (standard deviation <0.5%); however, any smaller permanent damage could not be assessed. PMID- 16696486 TI - A bench-top megavoltage fan-beam CT using CdWO4-photodiode detectors. II. Image performance evaluation. AB - Megavoltage computed tomography (MVCT) is a potential imaging tool for positioning and dose delivery verification during image guided radiotherapy. The problem with many MVCT detectors, however, is their low detective quantum efficiency (DQE) which leads to poor low contrast resolution (LCR) and high image noise. This makes separating the tumors from the soft tissue background difficult. This manuscript describes the imaging performance of our bench-top MVCT scanner that uses an 80-element detector array consisting of CdWO4 photodiode elements with a DQE of 19% in 6 MV and 26% in Co60 beams [T. T. Monajemi, S. Steciw, B. G. Fallone, and S. Rathee, "Modelling scintillator photodiodes as detectors for megavoltage CT," Med. Phys. 31, 1225-1234 (2004)] at zero frequency. The imaging experiments presented were carried out mainly in a Co60 teletherapy unit, while the beam hardening characteristics of the system were also presented for a 6 MV beam. During image evaluation, persistent ring artifacts, caused by air gaps at the ends of the eight-element detector blocks, were removed by using a calibration procedure. The measured contrast of a low contrast target with a 20 mm diameter was determined to be independent of dose, between 2.1 and 17 cGy. The measured LCR of a target with a nominal contrast of 2.8% was reduced from 2.3% to 1.2% when the contrast target diameter was reduced from 15 to 5 mm, using 17 cGy for imaging. The signal to noise ratio of this system is shown to be proportional to the square root of dose. Most importantly, a low contrast target with a diameter of 6 mm and a nominal contrast level of 1.5% is resolved with a radiation dose of 2.1 cGy in the Co60 beam. The spatial resolution in the Co60 beam is limited to one line pair per centimeter mainly due to the size of the Co60 source. PMID- 16696487 TI - AAPM TG-43U1 formalism adaptation and monte carlo dosimetry simulations of multiple-radionuclide brachytherapy sources. AB - This paper presents a preliminary study on multiple-radionuclide sources for brachytherapy. An adaptation of the AAPM TG-43U1 formalism is proposed in order to derive the dosimetry parameters of multiple-radionuclide sources from mono radionuclides. The adapted formalism is applied to a bi-radionuclide case with the help of Monte Carlo calculations (MCNPX 2.5.0). InterSource seed loaded with 103Pd and 125I was chosen. This combination promotes a higher dose rate than InterSource125 (loaded with 125I) and deeper tissue penetration than InterSource103 (loaded with 103Pd) while reducing the dose at long distance (beyond 2.5 cm) relative to InterSource125. In conclusion, this work shows the benefits of combining different radionuclides inside the same seed and proposes an adaptation of the AAPM TG-43U1 formalism for the implementation of multiple radionuclide sources in current treatment planning systems. PMID- 16696488 TI - Effect of scatter and an antiscatter grid on the performance of a slot-scanning digital mammography system. AB - The use of a grid increases perceptibility of low contrast objects in mammography. Slot-scan mammography provides a more dose efficient reduction of the scattered radiation reaching the detector than obtained with an antiscatter grid in screen-film or flat-panel digital mammography. In this paper, the potential of using a grid in a slot-scan system to provide a further reduction of scattered radiation is investigated. The components of the digital signal: primary radiation, off-focus radiation, scattered radiation, and optical fluorescence glare in a CsI(Tl) detector were quantified. Based on these measurements, the primary and scatter transmission factors (Tp, Ts), scatter-to primary ratio (SPR), signal-difference-to-noise ratio (SDNR), and the SDNR improvement factor (K(SDNR)) were obtained. Our results showed that the SPR ranged from 0.05 to 0.19 for breast thicknesses between 2 and 8 cm, respectively. The values of K(SDNR) ranged from 0.85 to 0.94. Because the slot-scanning system has an inherently low SPR, the increase in dose required when the grid is used outweighs the benefit of the small increase in SDNR. It is possible that greater benefit could be achieved by using a grid with a higher Tp, such as obtained using air-core technology. PMID- 16696489 TI - Accurate skin dose measurements using radiochromic film in clinical applications. AB - Megavoltage x-ray beams exhibit the well-known phenomena of dose buildup within the first few millimeters of the incident phantom surface, or the skin. Results of the surface dose measurements, however, depend vastly on the measurement technique employed. Our goal in this study was to determine a correction procedure in order to obtain an accurate skin dose estimate at the clinically relevant depth based on radiochromic film measurements. To illustrate this correction, we have used as a reference point a depth of 70 micron. We used the new GAFCHROMIC dosimetry films (HS, XR-T, and EBT) that have effective points of measurement at depths slightly larger than 70 micron. In addition to films, we also used an Attix parallel-plate chamber and a home-built extrapolation chamber to cover tissue-equivalent depths in the range from 4 micron to 1 mm of water equivalent depth. Our measurements suggest that within the first millimeter of the skin region, the PDD for a 6 MV photon beam and field size of 10 x 10 cm2 increases from 14% to 43%. For the three GAFCHROMIC dosimetry film models, the 6 MV beam entrance skin dose measurement corrections due to their effective point of measurement are as follows: 15% for the EBT, 15% for the HS, and 16% for the XR-T model GAFCHROMIC films. The correction factors for the exit skin dose due to the build-down region are negligible. There is a small field size dependence for the entrance skin dose correction factor when using the EBT GAFCHROMIC film model. Finally, a procedure that uses EBT model GAFCHROMIC film for an accurate measurement of the skin dose in a parallel-opposed pair 6 MV photon beam arrangement is described. PMID- 16696490 TI - Assessment of a new multileaf collimator concept using GEANT4 Monte Carlo simulations. AB - The aim of the work was to investigate in advance the dosimetric properties of a new multileaf collimator (MLC) concept with the help of Monte Carlo (MC) simulations prior to the production of a prototype. The geometrical design of the MLC was implemented in the MC code GEANT4. For the simulation of a 6 MV treatment beam, an experimentally validated phase space and a virtual spatial Gaussian shaped model placed in the origin were used. For the simulation of the geometry in GEANT4, the jaws and the two leaf packages were implemented with the help of computer-aided design data. First, transmission values for different tungsten alloys were extracted using the simulation codes GEANT4 and BEAMnrc and compared to experimental measurements. In a second step, high-resolution simulations were performed to detect the leakage at depth of maximum dose. The 20%-80% penumbra along the travel direction of the leaves was determined using 10 x 10 cm2 fields shifted along the x- and y-axis. The simulated results were compared with measured data. The simulation of the transmission values for different tungsten alloys showed a good agreement with the experimental measurements (within 2.0%). This enabled an accurate estimation of the attenuation coefficient for the various leaf materials. Simulations with varying width of the spatial Gaussian distribution showed that the leakage and the penumbra depend very much on this parameter: for instance, for widths of 2 and 4 mm, the interleaf leakage is below 0.3% and 0.75%, respectively. The results for the leakage and the penumbra (4.7+/ 0.5 mm) are in good agreement with the measurements. This study showed that GEANT4 is appropriate for the investigation of the dosimetric properties of a multileaf collimator. In particular, a quantification of the leakage, the penumbra, and the tongue-and-groove effect and an evaluation of the influence of the beam parameters such as the width of the Gaussian distribution was possible. PMID- 16696491 TI - Automated detection of lung nodules in CT scans: false-positive reduction with the radial-gradient index. AB - We present a number of approaches based on the radial gradient index (RGI) to achieve false-positive reduction in automated CT lung nodule detection. A database of 38 cases was used that contained a total of 82 lung nodules. For each CT section, a complementary image known as an "RGI map" was constructed to enhance regions of high circularity and thus improve the contrast between nodules and normal anatomy. Thresholds on three RGI parameters were varied to construct RGI filters that sensitively eliminated false-positive structures. In a consistency approach, RGI filtering eliminated 36% of the false-positive structures detected by the automated method without the loss of any true positives. Use of an RGI filter prior to a linear discriminant classifier yielded notable improvements in performance, with the false-positive rate at a sensitivity of 70% being reduced from 0.5 to 0.28 per section. Finally, the performance of the linear discriminant classifier was evaluated with RGI-based features. RGI-based features achieved a substantial improvement in overall performance, with a 94.8% reduction in the false-positive rate at a fixed sensitivity of 70%. These results demonstrate the potential role of RGI analysis in an automated lung nodule detection method. PMID- 16696492 TI - Robust frameless stereotactic localization in extra-cranial radiotherapy. AB - In the field of extra-cranial radiotherapy, several inaccuracies can make the application of frameless stereotactic localization techniques error-prone. When optical tracking systems based on surface fiducials are used, inter- and intra fractional uncertainties in marker three-dimensional (3D) detection may lead to inexact tumor position estimation, resulting in erroneous patient setup. This is due to the fact that external fiducials misdetection results in deformation effects that are poorly handled in a rigid-body approach. In this work, the performance of two frameless stereotactic localization algorithms for 3D tumor position reconstruction in extra-cranial radiotherapy has been specifically tested. Two strategies, unweighted versus weighted, for stereotactic tumor localization were examined by exploiting data coming from 46 patients treated for extra-cranial lesions. Measured isocenter displacements and rotations were combined to define isocentric procedures, featuring 6 degrees of freedom, for correcting patient alignment (isocentric positioning correction). The sensitivity of the algorithms to uncertainties in the 3D localization of fiducials was investigated by means of 184 numerical simulations. The performance of the implemented isocentric positioning correction was compared to conventional point based registration. The isocentric positioning correction algorithm was tested on a clinical dataset of inter-fractional and intra-fractional setup errors, which was collected by means of an optical tracker on the same group of patients. The weighted strategy exhibited a lower sensitivity to fiducial localization errors in simulated misalignments than those of the unweighted strategy. Isocenter 3D displacements provided by the weighted strategy were consistently smaller than those featured by the unweighted strategy. The peak decrease in median and quartile values of isocenter 3D displacements were 1.4 and 2.7 mm, respectively. Concerning clinical data, the weighted strategy isocentric positioning correction provided the reduction of fiducial registration errors, featuring up to 61.7% decrease in median values (versus 46.8% for the unweighted strategy) of initial displacements. The weighted strategy proved high performance in minimizing the effects of fiducial localization errors, showing a great potential in improving patient setup. The clinical data analysis revealed that the application of a robust reconstruction algorithm may provide high-quality results in patient setup verification, by properly managing external fiducials localization errors. PMID- 16696493 TI - Method for the evaluation of a average glandular dose in mammography. AB - This paper concerns a method for accurate evaluation of average glandular dose (AGD) in mammography. At different energies, the interactions of photons with tissue are not uniform. Thus, optimal accuracy in the estimation of AGD is achievable when the evaluation is carried out using the normalized glandular dose values, g(x,E), that are determined for each (monoenergetic) x-ray photon energy, E, compressed breast thickness (CBT), x, breast glandular composition, and data on photon energy distribution of the exact x-ray beam used in breast imaging. A generalized model for the values of g(x,E) that is for any arbitrary CBT ranging from 2 to 9 cm (with values that are not whole numbers inclusive, say, 4.2 cm) was developed. Along with other dosimetry formulations, this was integrated into a computer software program, GDOSE. FOR, that was developed for the evaluation of AGD received from any x-ray tube/equipment (irrespective of target-filter combination) of up to 50 kVp. Results are presented which show that the implementation of GDOSE. FOR yields values of normalized glandular dose that are in good agreement with values obtained from methodologies reported earlier in the literature. With the availability of a portable device for real-time acquisition of spectra, the model and computer software reported in this work provide for the routine evaluation of AGD received by a specific woman of known age and CBT. PMID- 16696494 TI - Image registration with auto-mapped control volumes. AB - Many image registration algorithms rely on the use of homologous control points on the two input image sets to be registered. In reality, the interactive identification of the control points on both images is tedious, difficult, and often a source of error. We propose a two-step algorithm to automatically identify homologous regions that are used as a priori information during the image registration procedure. First, a number of small control volumes having distinct anatomical features are identified on the model image in a somewhat arbitrary fashion. Instead of attempting to find their correspondences in the reference image through user interaction, in the proposed method, each of the control regions is mapped to the corresponding part of the reference image by using an automated image registration algorithm. A normalized cross-correlation (NCC) function or mutual information was used as the auto-mapping metric and a limited memory Broyden-Fletcher-Goldfarb-Shanno algorithm (L-BFGS) was employed to optimize the function to find the optimal mapping. For rigid registration, the transformation parameters of the system are obtained by averaging that derived from the individual control volumes. In our deformable calculation, the mapped control volumes are treated as the nodes or control points with known positions on the two images. If the number of control volumes is not enough to cover the whole image to be registered, additional nodes are placed on the model image and then located on the reference image in a manner similar to the conventional BSpline deformable calculation. For deformable registration, the established correspondence by the auto-mapped control volumes provides valuable guidance for the registration calculation and greatly reduces the dimensionality of the problem. The performance of the two-step registrations was applied to three rigid registration cases (two PET-CT registrations and a brain MRI-CT registration) and one deformable registration of inhale and exhale phases of a lung 4D CT. Algorithm convergence was confirmed by starting the registration calculations from a large number of initial transformation parameters. An accuracy of approximately 2 mm was achieved for both deformable and rigid registration. The proposed image registration method greatly reduces the complexity involved in the determination of homologous control points and allows us to minimize the subjectivity and uncertainty associated with the current manual interactive approach. Patient studies have indicated that the two-step registration technique is fast, reliable, and provides a valuable tool to facilitate both rigid and nonrigid image registrations. PMID- 16696495 TI - Using a gradient plot of luminance response curve as a metric for assessing and comparing softcopy display system quality. PMID- 16696496 TI - The flu...and you. AB - Seasonal influenza is primarily an upper respiratory illness that recurs yearly in slightly different forms. The "flu shot," or yearly vaccine, is the best precaution for preventing flu. All health care workers at risk are advised to be vaccinated every year. Occasionally more dangerous or even deadly forms develop, possibly leading to epidemics and pandemics. Both type A and B can cause epidemics, but influenza type A has greater potential to develop new and deadly subtypes and is responsible for the current "bird flu" that threatens to mutate to a form capable of a human pandemic. The flu is spread by droplets of oral and respiratory secretions. In dental facilities, in addition to Standard Precautions, Droplet Precautions should be observed when people with highly contagious respiratory infections such as influenza are present. Consistent use of masks and careful hand hygiene, including use of alcohol-based waterless agents in both treatment and public areas by both employees and patients, are the most important precautions to control influenza. CDC Respiratory Hygiene/Cough Etiquette (available in posters from the CDC) is the perfect starting point to inform workers and patients how to keep the dental office "a safe place to be healed" rather than a place to contract the flu. PMID- 16696497 TI - The office environment: take a good look. PMID- 16696498 TI - The value of exit interviews. PMID- 16696499 TI - Informed consent: why you need more than a signature. PMID- 16696501 TI - Dental exams become field duty. PMID- 16696500 TI - The role of the Federal Trade Commission in advertising health products and services. AB - The Federal Trade Commission plays a unique role in enforcing well-established standards ensuring that consumers can make informed purchase and use decisions about health-related products and services based on truthful, nonmisleading advertising claims. Deceptive and unfair practices are defined. The importance of the "net impression" that ads convey to consumers and the need for substantiation of objective, factual claims is explained. The FTC uses its enforcement powers and consumer and industry outreach to create a climate for preventing misleading advertising. PMID- 16696502 TI - Your gift--to yourself and to your patient. PMID- 16696503 TI - Code smart, collect sooner. PMID- 16696504 TI - Clinical governance in Pakistan: myth or reality? PMID- 16696505 TI - Septic arthritis in a tertiary care hospital. AB - OBJECTIVES: To study the epidemiological features of septic arthritis in the adult population and to identify the risk factors for mortality in septic arthritis. METHODS: A five year reterospective study was performed on cases with septic arthritis admitted in our hospital between January 1999 and December 2004. Patients were identified according to ICD codes, 711.00. Data was recorded on a standardized data sheet and analyzed by SPSS 11.5 software. RESULTS: A total of 116 patients were identified, 69 were male (59.5%) and 47 female (40.55%). Mean age of patients was 49.22 years. The most common presenting clinical features were joint swelling (99.1%) and fever (60.3%), Knee joint was the most common joint involved (65.5%) followed by hip (11.2%). Gram stains of synovial fluid was done in 67.2% of cases out of which 22.4% had positive stains. Staphylococcus aureus was the most common organism isolated from blood as well as synovial fluid (18.8%). Mean haemoglobin was 10.83gm/dl and 57.8% of patients had total leukocyte count less than 11,000/cumm. Platelet count was greater than 150,000/cumm in 90.5% patients. Hypertension, renal failure, chronic liver disease and elevated ESR were identified as some of the potential risk factors for higher mortality in a cohort with septic arthritis. CONCLUSION: Septic arthritis is associated with significant morbidity and mortality. These results highlight the importance of obtaining cultures before starting any treatment. PMID- 16696506 TI - Frequency and determinants of vaginal infection in postpartum period: a cross sectional survey from low socioeconomic settlements, Karachi, Pakistan. AB - OBJECTIVE: To determine the frequency and factors associated with perceived vaginal infections among married women in their postpartum period. METHODS: A cross-sectional study was conducted from July 2000 to October 2000 in five squatter settlements of Karachi, Pakistan. These squatter settlements were selected on the basis of an existing surveillance system run by female community health workers for maternal and child healthcare which identified women who had delivered 42-56 days prior to the date of interview. Vaginal infection was considered present when a mother perceived foul smelling vaginal discharge during the postpartum period. Mothers were interviewed to gain insight into socioeconomic and demographic variables, materials used to staunch lochia, duration of labour, personal and perineal hygiene and past obstetric history. RESULTS: A total of 525 women were interviewed. The estimated prevalence of perceived vaginal infection was 5.1%. Factors associated with perceived vaginal infections included, delivery conducted by a non-medical personnel (AOR 3.5, CI 1.3-9.5) and use of unhygienic cloth or cotton for staunch of lochia (AOR 2.7, CI 1.1-6.2). CONCLUSION: Among women who reported perceived vaginal infection, a higher proportion were delivered by non-medical personnel, and used unhygienic material (cloth or cotton) for staunch of lochia as compared to women who did not perceive vaginal infection. We recommend deliveries to be conducted by trained personnel and provision of health education for persons who conduct delivery and women to use hygienic material for staunch of lochia during post partum period. PMID- 16696507 TI - Diagnostic value of chest X-ray and echocardiography for cardiac tamponade in post cardiac surgery patients. AB - OBJECTIVE: To investigate the effectiveness of X-ray and echocardiography (ECHO) as a diagnostic tool for cardiac tamponade in adult cardiac surgery patients. METHODS: Thirty five coronary artery bypass and graft surgery patients who developed cardiac tamponade at Cleveland clinic foundation, were included in this study. Their diagnosis was confirmed at the time of re-exploration in the operating room. These patients were followed retrospectively for demographics, X ray and echocardiography findings. Abnormal and enlarged cardiac silhouette which was different from first postoperative X-ray was used as radiological criteria for tamponade while echocardiographic diagnosis was left to the discretion of cardiologist. RESULTS: Twenty four males (69%) and 11 (31%) females with an average age of 60.7 +/- 15.2 years were included in this study. Only 7 (20%) patients met our criteria for radiological diagnosis of tamponade. All 35 patients showed pericardial effusion on ECHO while cardiac tamponade was present in 30 (86%) patients. Tamponade developed on an average on 4th postoperative day. CONCLUSION: Radiological diagnosis of cardiac tamponade based on changes in cardiac silhouette provide limited information while ECHO is a reliable diagnostic tool when combined with clinical findings. PMID- 16696508 TI - Unplanned prolonged postanaesthesia care unit length of stay and factors affecting it. AB - OBJECTIVE: To identify the factors that prolong the length of stay inthe post anaesthesia care unit (PACU). METHODS: This audit was conducted in the PACU of a university hospital. A special form was designed and filled for those patients who stayed unplanned in the PACU for more than two hours. All patients who were admitted to the PACU after surgery were included. Patients undergoing cardiothoracic surgery, those directly shifted to ICU and cases done under local anaesthesia were excluded. Data was collected for 20 months by a designated recovery nurse for all included patients including those admitted outside the scheduled surgery hours. RESULTS: The total number of patients who were admitted to the PACU during the audit period were 13644, out of these 1114 (8.1%) stayed in the PACU for more than 2 hours. The percentage of overstay patients on monthly basis ranged from 6.4% to 10%. The commonest reason was the need for postoperative monitoring 578 (51.8%), unavailability of beds in the special care areas 264 (23.7%), pain management 68 (6.1%) and 61 (5.4 %) for postoperative ventilation. CONCLUSION: Our results show that majority of patients stayed in the PACU for more than two hours either because they needed postoperative monitoring or because of unavailability of bed in the special care areas. PMID- 16696509 TI - Standardization of Anaesthesia Ready Time and reasons of delay in induction of anaesthesia. AB - OBJECTIVE: Anaesthesia-Ready Time (ART) is the time taken by the anaesthetist to provide sufficient anaesthetic depth for start of surgery. Our aim was to set benchmark timings for ART and compare it with our current practice. METHODS: Benchmark ART time of 15 minutes was set for American Society of Anesthesiologists (ASA) class I and II patients, 30 minutes for ASA III and IV patients, 20 minutes for spinal and 30 minutes for epidural anaesthesia. An additional 15 minutes was added for each invasive procedure. RESULTS: Three hundred elective cases were audited. Seventy eight percent of the cases were within benchmark timings. The main causes of delay included undergraduate students performing procedures (24.6%), teaching invasive lines to postgraduates (21.3%) and paediatric patients (16.4%). CONCLUSION: The introduction of benchmark timings and its regular auditing can help standardize operating room booking time and reducing patient cost. PMID- 16696510 TI - Tuberculous tenosynovitis: a cause of Carpal Tunnel Syndrome. AB - OBJECTIVE: To highlight the presentation of tuberculous tenosynovitis as Carpal Tunnel Syndrome (CTS). METHODS: A descriptive study conducted with purposive sampling on the patients presenting between April 2004 to January 2005 to the department of Plastic Surgery, CMH Rawalpindi, with the clinical picture of CTS. Symptoms and signs were recorded. The carpal tunnels were explored under nerve blocks. Where tenosynovitis was observed per-operatively, specimen of excised synovium were sent for histo-pathological examination, acid fast stains and bacterial cultures, to ascertain or rule out the presence of tuberculosis. RESULTS: A total of 53 patients predominantly males were included in the study. The median age was 43 years and average duration of symptoms was 9 months. Tenosynovitis was observed in three patients (6%) per-operatively, with histopathology suggestive of Tuberculous tenosynovitis. Point Prevalence of tuberculous tenosynovitis as a cause of CTS was 6%. These patients reported clinical improvement with anti-tuberculosis treatment. NCS/EMG co-related well with clinical relief. CONCLUSION: Tuberculous tenosynovitis is an uncommon cause of median nerve compression at the wrist. As symptoms are usually typical of CTS, diagnosis is frequently missed. Consideration of the possibility, examination of the opened canal at surgery and proper treatment can result in a successful outcome. PMID- 16696511 TI - Correlates of injection use of synthetic drugs among drug users in Pakistan: a case controlled study. AB - OBJECTIVE: To assess the various correlates of injection of synthetic drugs among drug users in Pakistan. METHODS: In a case controlled study, 141 cases and 272 controls were recruited, from 11 drug sites in Lahore, Pakistan, through multistage sampling techniques at a case control ratio of 1:2. Cases were defined as injecting drug users (IDU's) who switched from non injection routes of drug intake to injecting synthetic drugs, while controls (272) were heroin chasers who had never injected synthetic drugs. Data were collected on various demographic, socioeconomic and personal characteristics, drug use history and practices, and knowledge regarding HIV. While controlling for various confounders, adjusted odds ratios (aOR) with corresponding 95% CI were estimated using multivariate logistic regression. RESULTS: Factors found to be independently associated with route transitions were young age (aOR:3.7, 95%CI: 1.9-7.2), homelessness (aOR: 6.4, 95%CI: 3.9-10.2), presence of an IDU friend (aOR:3.7, 95%CI: 2.3-6.1), easy availability (aOR:5.6, 95%CI: 3.3-9.6) and cost of the current drug (aOR:2.6, 95%CI: 1.6-4.3) and poly drug use (aOR:3.2, 95%CI: 1.6-6.4). CONCLUSION: Low cost of the current drug of choice, easy availability and polydrug use were strongly associated with IDU of synthetic drugs. PMID- 16696512 TI - Pakistan--country profile of cancer and cancer control 1995-2004. PMID- 16696513 TI - Role of ursodeoxycholic acid in lowering ALT in chronic liver disease. AB - The objective was to see the efficacy of ursodeoxycholic acid in improving the ALT levels in patients suffering from chronic liver disease. Thirty patients suffering from chronic liver disease (either B or C) were given oral ursodeoxycholic acid in a dose of 250 mg twice a day for 4 months. Their blood biochemistry and haematology were repeated monthly for 7 months i.e. 4 months of therapy and 3 months post therapy to see the response. The mean ALT levels of 30 patients who completed the study was 101 +/- 47 IU/L. During therapy 24 cases (80%) showed lowering of their ALT levels while 6 (20%) either showed no response or worsening of ALT levels. Of 24 cases who showed an improvement in their ALT levels; over 25% drop in ALT levels was seen in 217 cases (70%) and less than 25% drop seen in 7 cases (30%). The mean ALT values during therapy were 75 IU/L showing an overall 25% reduction from the baseline values. Following cessation of therapy the mean ALT levels showed a rise to 90 IU/L which was almost similar to 101 IU/L value in the pre treatment period. The study showed some role of ursodeoxycholic acid in improving the ALT levels in chronic liver disease. PMID- 16696514 TI - Trans-mastoid approach to otogenic brain abscess. AB - The treatment of otogenic brain abscess initially involves excision or aspiration of the abscess through a temporal or sub-occipital route depending on its location. This is followed by a mastoidectomy by the ENT surgeon to eradicate the primary source of infection. During the last three years, we have approached such lesions through a mastoidectomy followed by excision of the abscess through the same approach. This trans-mastoid approach is technically feasible in following the tract of suppuration, and clearing the cause and effect of pathology, at the same sitting. This paper describes our initial experience with the trans-mastoid approach to otogenic brain abscesses. On the basis of our results, we believe that transmastoid approach is an effective and logical option for the treatment of otogenic brain abscess, and merits further investigation in the form of a prospective study. PMID- 16696515 TI - Management of compromised airway due to unusual presentation of cystic hygroma. AB - The successful management of a unique case of huge cystic hygroma in a fourteen month old child who presented in the emergency room with severe respiratory distress as a result of nearly almost complete upper airway obstruction, is presented. PMID- 16696516 TI - Follicular dendritic cell sarcoma of lymph node--a rare entity. AB - Follicular dendritic cells (FDC) are non-lymphoid, non-phagocytic accessory cells in the immune system that are essential for antigen presentation and germinal center reaction regulation. These cells are CD21+, CD35+, CD1a- and S100 protein +/- and they show desmosomes ultrastructurally. The most commonly involved sites by FDC tumors are lymph nodes but may arise at a variety of extranodal sites including oral cavity, tonsil, gastrointestinal tract and liver. Most studies represent single case reports or case series. Our patient presented with tumor in the lymph nodes. Histology revealed tumor cells with abundant eosinophilic cytoplasm, hyperchromatic and pleomorphic nuclei, and prominent nucleoli. The tumor cells were found to be positive for CD21 which is a specific marker for follicular dendritic cells. PMID- 16696517 TI - Unusual site of guide-wire entrapment during central venous catheterization. AB - We are reporting a case of possible entrapment of guide wire in the region of tricuspid valve, which made it impossible to remove the guide wire from internal jugular (IJ) vein for a short while. Awareness about this rare complication is emphasized. PMID- 16696518 TI - Life threatening mediastinal haematoma: a complication of central venous catheterization. AB - Central venous catheterization (CVC) has established risks and benefits in its application as a vascular access source, particularly in situations involving temporary cannulation. We present a rare case of life-threatening mediastinal haematoma resulting from CVC usage. Even though aggressive intervention yielded survival, the patient was left with permanent vocal compromise owing to traumatic palsy of the right recurrent laryngeal nerve. One should be careful in selection of venous access and be aware of alternatives routes. PMID- 16696519 TI - Frequency and sensitivity pattern of extended spectrum beta lactamases in a tertiary care hospital laboratory of Pakistan. PMID- 16696520 TI - The earthquake aftermath in Pakistan: unearthing the most vulnerable. PMID- 16696521 TI - Chronic obstructive pulmonary disease and cardiovascular disease. PMID- 16696522 TI - Inhaled formoterol versus ipratropium bromide in chronic obstructive pulmonary disease. AB - BACKGROUND: Short-acting anticholinergic bronchodilator, ipratropium bromide has been recommended as first-line drug in chronic obstructive pulmonary disease (COPD). More recently, long acting beta2-agonist (LABA) bronchodilators such as formoterol have been shown to be useful in COPD. Limited information is available on the relative efficacy of these two drugs in COPD. METHODOLOGY: A randomised, double-blind, cross-over, placebo-controlled study was carried out. Forty-four stable patients with COPD received single doses of formoterol (12 microg), ipratropium bromide (40 microg) or placebo, administered through a metered-dose inhaler on three consecutive days in a random order. Spirometry, static lung volumes, pulse rate and blood pressure, and assessment of sensation of dyspnoea at rest using a visual analog scale (Borg Scale) were recorded at baseline. Subsequently, these were repeated for assessment of response: spirometry at 5, 30 and 60 minutes and static lung volumes, pulse rate, blood pressure and dyspnoea measurement at 60 minute. RESULTS: Formoterol resulted in greater immediate improvement in lung function, with the change in FEV1 at 5 min being greater than that observed with ipratropium. The changes in static lung volumes were similar between the two but superior to placebo. Both the drugs reduced dyspnoea. Formoterol produced a significantly greater increase in heart rate and systolic blood pressure as compared to ipratropium, although the magnitude of these changes was small and clinically unimportant. CONCLUSIONS: Single therapeutic doses of formoterol and ipratropium bromide are equally effective in improving lung function and reducing dyspnoea. However, formoterol appears to be a better bronchodilator producing a faster improvement in lung function. PMID- 16696523 TI - Prediction model for peak expiratory flow in North Indian population. AB - OBJECTIVE: To establish a model for predicting peak expiratory flow rate (PEFR) in North Indian healthy population. Study subjects. Eight hundred and ninety seven healthy, non-smoker individuals (681 males and 216 females) in the age group of 10-60 years. METHODS: The study was carried out at a health exhibition organised by the Government of Uttar Pradesh, at King George's Medical University, Lucknow. Only healthy, non-smoker individuals were enrolled for the study. Age was noted in completed years and weight in kg and height in cm were taken without shoes. PEFR was measured by Mini Wrights peak flow meter in standing position after prior instructions and demonstration of technique to each individual. The test was performed three times on each subject and best of the three attempts was selected for data computation. The statistical solftware SPSS was used to fit the model and perform residual analysis. RESULTS: The highest reading for males was recorded in the age group of 20 to 24 years and for females in the age group of 25 to 29 years. Using age, height and weight, we established a regression model for predicting PEFR values for males and females separately in the age group 19-60 years. In the age group 10-18 years, the model for predicting PEFR was same for both the sexes. PEFR values were found to be more in males as compared to females. The predictive power of the model as described by explained variation was found to be 80 and 82 percent for males and females, respectively. CONCLUSIONS: Prediction model for north Indian subjects was drafted for age range 10-60 years. While separate models were required for males and females because of sex related differences in the age group 19-60 years, a common model sufficed for age group 10-18 years. PMID- 16696524 TI - Atypical mycobacterial infection among HIV seronegative patients in Pondicherry. AB - BACKGROUND: The role of non-tuberculous mycobacteria (NTM) as aetiological agents in the causation of lung infections has seldom been systematically studied from India. METHODS: Prospective study of Species identification from mycobacterial isolates. RESULTS: A total of 22 isolates of mycobacteria were isolated from 635 samples of suspected tuberculosis in a two months period, of which 17 were Mycobacterium tuberculosis, three Mycobacterium kansasii and one each of Mycobacterium fortuitum and Mycobacterium gordonae. CONCLUSIONS: This study highlights the importance of NTM as potential pathogens from both pulmonary and extrapulmonary sites in the Southern state of Pondicherry. Further studies on a larger scale are needed to delineate clearly the association of NTM and various infections, especially in immunocompromised patients. PMID- 16696525 TI - Clinical profile, diagnosis and management of patients presenting with symptomatic pulmonary embolism. AB - OBJECTIVE: To study the clinical profile and diagnostic methods in patients with symptomatic pulmonary embolism (PE). METHODS: Prospective assessment of clinical features, radiology and outcome of patients presenting with symptomatic PE over an 18-month period. RESULTS: During study period, 24 patients with a mean age of 39 +/- 12.1 years were diagnosed to have symptomatic pulmonary embolism. Dyspnoea (91.7%) and cough (58.3%) were the predominant complaints. Spiral computed tomographic pulmonary angiography (CTPA) was performed in 21 (87.5%) patients and perfusion scans in 14 (58.4%) patients. Echocardiography performed in all patients revealed evidence of pulmonary artery hypertension and right ventricular dyskinesia in 20 (83.3%) and 15 (62.5%) patients, respectively. Thrombolysis with streptokinase was performed in 14 (58.3%) patients. All patients received low molecular weight heparin followed by warfarin. Of the 24 patients, 20 (83.3%) were discharged and are under regular follow-up; four patients died. CONCLUSIONS: Pulmonary embolism is a common problem and can be easily diagnosed provided it is clinically suspected. Early diagnosis and aggressive management is the key to successful outcome. PMID- 16696526 TI - Hypersensitivity pneumonitis. AB - Hypersensitivity pneumonitis (HP) or extrinsic allergic alveolitis is a non-IgE mediated hypersensitivity disease initiated by inhalation and subsequent sensitisation to organic antigens. These diseases have been described in different occupational groups and present in acute, subacute or chronic forms based on the exposure to antigens and host response. Clinical features are dependent upon the stage of the disease and can include fever, chills, cough, dyspnoea, and weight loss. The immunopathogenesis involves both cellular immunity and antibody responses to inhaled antigens. Antibody response to the implicated antigen can be demonstrated in HP patients, but such antibodies are also detected in antigen exposed asymptomatic individuals. Bronchoalveolar lavage demonstrates lymphocytosis and preponderance of CD8+ cells. Pulmonary function studies demonstrate a restrictive pattern with diffusion defects. The diagnosis is difficult as no single test is confirmatory, hence information from clinical, radiological, physiological, and immunological evaluations may be used together for a confirmative diagnosis of hypersensitivity pneumonitis. The treatment of choice is avoidance of antigen but systemic corticosteroids may be effective in suppressing the inflammatory response. The prognosis depends on early diagnosis and effective antigen avoidance. PMID- 16696527 TI - Apical mass presentation of brachial plexus Schwannoma. AB - Neurogenic tumours arising from the brachial plexus are rare. Most of these present as palpable cervical mass with tenderness and pain radiating to the arm along the distribution of the affected nerve. Here we report a case with radiological presentation mimicking an apical mass in the right chest with minimal symptoms. The mass was successfully and completely removed at thoracotomy preserving the integrity of the lower trunk of the brachial plexus. There was some evidence of neuroparesis in the immediate post-operative period but the patient made a very good recovery 10 weeks after surgery. PMID- 16696528 TI - Cardiac sarcoidosis: an uncommon presentation of sarcoidosis in India. AB - Cardiac involvement in sarcoidosis is uncommon. The manifestations of cardiac sarcoidosis are protean and vary from benign ectopics to life threatening ventricular tachycardias. High index of suspcion and early aggressive intervention is required for the diagnosis and treatment of this condition. Early diagnosis is essential since ventricular tachycardia is the most common terminal event in these patients if left untreated. Corticosteroids are needed early in these patients which along with other antiarrhythmic management strategies including intracardiac defibrillator form the backbone of treatment. We describe four patients who were diagnosed to have pulmonary and cardiac sarcoidosis. Their varied presentations highlight the difficulties that may occur in the diagnosis. Furthermore, the need for proper management is stressed, as a delay in diagnosis may be fatal. PMID- 16696529 TI - Primary pulmonary Hodgkin's disease: a distinct entity. AB - A 30-year-old lady presented with fever, dry cough and weight loss for the preceding five months. Radiological investigations revealed a solitary nodular lesion in the lingula of the left lung. Guided fine needle aspiration cytology failed to yield any diagnostic material. Bronchoscopic cytology was also not contributory. As a last resort open lung biopsy was done and a diagnosis of Hodgkin's disease was made. Hilar and pre aortic lymph node biopsies showed only reactive change. The final diagnosis was primary pulmonary Hodgkin's disease. PMID- 16696530 TI - Idiopathic endogenous lipoid pneumonia. AB - Lipoid pneumonia is a rare pulmonary disorder having no classical radiological appearance. We report a 33-year-old male, ex-smoker who was referred to us with history of cough, mild mucoid expectoration and progressively increasing dyspnoea since one year. He was investigated at local hospital and was treated with 30 mg prednisolone per day for 6 months for sarcoidosis without any response. On examination, he was normal except for fine basal crepitations in chest. Pulmonary function test (PFT) revealed mild airway obstruction. High resolution computerised tomographic scan (HRCT scan) revealed bilateral reticulonodular shadows and bronchiectasis in lower zones. Open lung biopsy revealed lipoid pneumonia. As there was no history of nasal distillation of oils, it was diagnosed to be idiopathic. The relevant literature is reviewed. PMID- 16696531 TI - A case report of pulmonary placental transmogrification with review of literature. AB - Pulmonary placental transmogrification (PPT), a rare cystic lesion of the lung is described in a 38-year-old male who had no risk factor for emphysema. The lesion was characterised by large bullae with its inner configuration resembling placental chorionic villi macroscopically and merging imperceptibly with adjacent normal lung tissue. To the best of our knowledge, this is the first case report from India and South Asia. Awareness of this rare entity helps make a definite diagnosis. Surgery alone is curative and not conservative management as prescribed for emphysema. PMID- 16696532 TI - Oswego Hospital powers up with modular system. PMID- 16696533 TI - The future is now. Flexibility and expandability drive hospital project. PMID- 16696534 TI - Wrapping it up. The hospital's first line of defense against moisture. PMID- 16696535 TI - The test of time. As health care evolves, so does Planetree. PMID- 16696536 TI - Planning for power. An electrical system designed for current and future demands. PMID- 16696537 TI - Re-raising the bar. ASHE takes its construction certificate to the next level. PMID- 16696538 TI - Moving up. Where can ES managers find help with their transport duties? PMID- 16696539 TI - Multi-cultural and multi-ethnic considerations and advanced directives: developing cultural competency. AB - Attention to culturally specific rituals germane to end-of-life rituals are important for the nurse who is delivering culturally competent care. The Patient Self-Determination Act implemented in the United States in 1991 brought with it some specific assumptions of values related to end-of-life care involving patient autonomy, informed decision making, truth telling and control over the dying process (USC, 1990). The assumptions and values assumed in this act are not necessarily shared by persons in the United States who are from a minority culture, particularly one other than White of European descent. This manuscript considers the values of the Patient Self-Determination Act passed in the United States and specific cultural beliefs which may pose a conflict for the care giver in the delivery of culturally competent care. The Giger and Davdihizar (2004) Transcultural Assessment Model is used as a way to assess patient and the family in an effort to plan culturally appropriate care that recognizes the uniqueness of each individual at the end of life. PMID- 16696540 TI - Enhancing Success in Advanced Practice Nursing: a grant-funded project. AB - The Enhancing Success in Advanced Practice Nursing (ESAPN) Project is designed to improve access to a diverse and culturally competent and sensitive health professions workforce by increasing the number of Hispanic, African-American and Asian nurses recruited, enrolled in and graduated from the MSN program at the University of Medicine and Dentistry of New Jersey-School of Nursing (UMDNJ-SN). In addition, the project plan includes the development and implementation of a comprehensive program that incorporates academic support services, career advisement and mentoring activities to retain and graduate an increased number of culturally, racially and ethnically diverse advanced practice nurses for the State of New Jersey. The project also seeks to improve the quality of care by preparing advanced practice nurses to provide culturally competent and sensitive care by assuring that the MSN curriculum includes content and clinical experiences relevant to the development of cultural competence. Faculty participation in workshops, designed to increase knowledge of cultural competence, is a key component. The success of the project will be evaluated using a variety of measures that track increases in the number of diverse students recruited and enrolled, the number of students accessing services associated with the ESAPN program, and increased retention and graduation of Hispanic, African-American and Asian nurses prepared as advanced practice nurses. PMID- 16696541 TI - International immersion programs in baccalaureate nursing education: professor and student perspectives. AB - This article presents the dual perspectives of professor and student in relation to the explicit need for nurses to become acutely conscious of cultural competence through self-awareness and how this influences nursing care. Mary Anne Levine, Professor of Nursing, discusses the development, planning, implementation, and evolution of the International Immersion Program (IIP) at Humboldt State University. The student experience, integration, and evaluation of IIP and its impact on one's professional and personal life are described by Elizabeth Perpetua, currently a practicing RN, who participated in the program in Nakhodka, Russia. PMID- 16696542 TI - Prerequisites to providing culturally competent care to Mexican migrant farmworkers: a Delphi study. AB - The purpose of this study was to identify prerequisites to providing culturally competent care to Mexican migrant farmworkers. A national sample of nurse experts who worked with Mexican migrant farmworkers (N = 93 [50.2%] in round 1; and N = 142 [54.8%] in round 2) participated during the period of August 1998 to April 1999. Using a two-round modified Delphi method, a list of 89 items was generated. The nurse experts agreed that the 89 items identified were prerequisites to cultural competence, showing consensus rates of 85.4% in round 1 and 88.8% in round 2. The prerequisites to culturally competent care identified by participants encompassed caring, cultural sensitivity, cultural knowledge (general/specific), cultural abilities/skills, and others (bi-cultural status; cultural and learning experiences). PMID- 16696543 TI - Nepalese school-age children's self-reported fears and coping strategies related to medical experiences. AB - The purpose of this study was to determine the fears of healthy Nepalese school age children and their coping strategies relative to medical experiences. The sample consisted of 161 children from a private and a public school in Nepal. Consistent with cognitive developmental theory and coping theory, Nepalese school age children were capable of reporting their fears of medical experiences and their coping strategies. Nepalese school-age children reported relatively moderate levels of fears and limited use of coping strategies. A significant difference was found between boys and girls self-reported fears (t=1.94, p=.05). There was no significant difference in children's fears based on the type of school attended. However there was a significant difference between public and private school children's report of frequently used coping strategies (t=4.10, p=. 000) and coping effectiveness (t=4.30, p=. 000). Nursing implications are discussed. PMID- 16696544 TI - Identity and health in the narratives of older mixed ancestry Asian Americans. AB - The United States has experienced rapid growth of the Asian American population in the last decade. People of mixed ancestry are a significant proportion of Asian America. Little is known about the health beliefs and health practices of this extremely diverse population. Thirteen older racially mixed Asian Americans, ranging in age from 48-94, were interviewed in a qualitative study that included questions about identity, health beliefs, and health practices. Narrative analysis revealed a relationship between identity, health practices, and interpretation of experiences with health care providers. PMID- 16696545 TI - Immersion in another culture: one strategy for increasing cultural competency. AB - Educating nursing students in the United States to be culturally competent is a challenge. Undergraduate and graduate nursing students, including nurse-midwifery students were paired with faculty on a short-term transcultural nursing experience in Choluteca, Honduras. Students provided health screenings, prenatal assessments, and birth attendance. Learning objectives were designed to improve cultural competency in the healthcare setting for nursing students. A case study exemplar in the labor and delivery setting is presented. As a result of the cultural immersion experience, the student demonstrates versatility and flexibility in clinical practice, reflecting the skills necessary to adapt interventions to populations and circumstances. PMID- 16696546 TI - Cultural/interpersonal values and smoking in an ethnically diverse sample of Southern California adolescents. AB - In ethnically diverse school contexts, values from multiple cultures might influence adolescents' attitudes and behaviors. This study developed scales to assess cultural values among Southern California 6'-grade adolescents (N=2281) and evaluated the associations between values and smoking. The scales assessed values salient in many Hispanic and Asian cultures: Respect for Adults (e.g., filial piety, respeto), Interpersonal Harmony (e.g., saving face, simpatia), and Differentiated Gender Roles (e.g., machismo). In cross-sectional and one-year longitudinal models, Respect for Adults and Interpersonal Harmony were associated with a lower risk of lifetime smoking. The associations were significant even after controlling for demographic characteristics, friends' smoking, and parents' smoking, indicating that values influence adolescents' behavior over and above the effects of modeling and peer influence. Increased understanding of adolescents' values could inform the creation of smoking prevention programs for ethnically diverse adolescents. PMID- 16696547 TI - Symmetry types, systems and their multiplicity in the structure of adenovirus capsid. I. Symmetry networks and general symmetry motifs. AB - Each of the more than 1500 polypeptide molecules of 7 different types building up the adenovirus capsid--probably even those of their amino-acids--are in symmetrical location. Every kind of polypeptide forms a separately also symmetrical network in the capsid distributed according to their functions in the inner and outer side and the inside of the facets and edges, but always in compliance with the icosahedral symmetry. Therefore, each different polypeptide also means a general symmetry motif in the capsid in its own symmetry network. Hexons can be considered as general symmetry motifs in some special association that is because of their environmental position four kinds of hexon types can be found, which are on every facet, next to one another, like three identical groups of four (GOF) according to the three-fold rotational symmetry. Two polypeptides of a peripentonal hexon of each GOF orient toward the penton and the third toward the other penton located further on the same edge. There are two versions of the arrangement of the GOFs: the hexons surround either a polypeptide IX or a polypeptide IlIa. The two versions of GOFs on 20 facets symmetrically recurring 60 times as general hexon symmetry motifs form the capsid in combination with the network of other polypeptides. Ideally, the surface of the hexon trimer shows three-fold rotational and three-fold reflexional symmetries. In the arrangement of hexons in the facets the translational, rotational, horizontal and vertical reflexional symmetry and the combination of these, as well as the glide reflexion and the antisymmetry can be found. Each hexon has six nearest neighbours and every hexon takes part in the construction of three hexon rows. Every facet and every vertex made up of five facets has an antisymmetrical pair located on the opposite side of the capsid. Every triangular facet participates in forming three vertices and every facet has three nearest neighbouring facets. In the facets, the polypeptide subunits of polypeptide IX centered GOF hexons have identical counter-clockwise orientation but the orientation of the neighbouring facets is always opposite compared to each other. On the five-fold symmetry axis, any facet can be "turned on" to the adjacent facet or "rotated" to all the others and will take the symmetry and orientation of the facet it got turned on or rotated to. Thus, every facet together with the polypeptides attached to it shows a twenty fold symmetry and multiplicity. An other type of symmetry and multiplicity in the capsid is that perpendicular to the 6 five-fold rotation axes run a geodetic (equatorial) ribbon like motif (superfieces) altogether six made up of 10 x 10 triangular facets and bent ten-times with an angle of 36 degrees. A triangular facet participates in forming three ribbon-like motifs, which intersect with each other on the given facet, but the same three motifs intersect repeatedly only on the antisymmetrically located facet. PMID- 16696548 TI - Prevalence and avidity of human herpesvirus-6 specific IgG antibodies in pregnant women in Hungary. AB - The prevalence, the level and the avidity of human herpesvirus-6 (HHV-6) specific IgG were examined in pregnant women and age-matched female blood donors. The study group consisted of 180 women (age 14-45); 60 women with normal pregnancy, 60 pregnant women with fetuses suspected of having any viral infection and 60 healthy blood donors with no history of pregnancy. Plasma or serum samples were tested for HHV-6 IgG antibodies by an immunofluorescence assay. Ninety-eight percent of blood donors and 97% of 120 pregnant women had IgG antibodies to HHV 6. The rate of seropositivity in women with normal pregnancies and women with fetuses suspected to have viral infection was the same. Pregnant women (n = 120) had significantly lower antibody titer than blood donors. No significant differences were found in the same respect between the two groups of pregnant women. Low avidity of IgG antibodies to HHV-6 was detected in 5% of pregnant women. PMID- 16696549 TI - Chlamydophila pneumoniae and human cytomegalovirus in atherosclerotic carotid plaques--combined presence and possible interactions. AB - The aim of our study was to investigate the combination of Chlamydophila pneumoniae and human cytomegalovirus (HCMV) as a pathogenic factor in atherosclerosis. Accordingly, we tested by means of PCR and immunohistochemistry the presence of these pathogens in the same atherosclerotic carotid specimen. The histology of the samples and the patients' antibodies against these pathogens were evaluated. Further, we examined the impact of C. pneumoniae and HCMV infection on the gene expression of the human monocytic cell line U937. Six of the 22 samples contained only C. pneumoniae, 4 contained only HCMV, 7 contained both C. pneumoniae DNA and/or antigens of both pathogens, and 5 samples were negative. No correlation was found between the presence of these microbes and either the cellular structure of the plaques, or the serostatus of the patients. The infection of U937 cells with HCMV and especially C. pneumoniae induced inflammation and atherosclerosis-related genes. Furthermore, the doubly-infected cells produced higher levels of the mRNA of pro-platelet basic protein and fatty acid binding protein 4. In conclusion, C. pneumoniae is often present in combination with HCMV in atherosclerotic carotid lesions. The in vitro coinfection model reveals that the doubly-infected monocytes are potent expressors of proatherosclerotic genes, suggesting that this coinfected population may accelerate the process of atherosclerosis. PMID- 16696550 TI - Nef: a pleiotropic modulator of primate lentivirus infectivity and pathogenesis. AB - The lentiviral protein Nef recruits cellular signalling proteins to lipid rafts at the cell membrane and acts thereby as a master regulator affecting the transcription of a series of cellular genes. By activating resting T cells, Nef creates an optimal environment for lentivirus replication. In human immunodeficiency virus (HIV) infected macrophages and microglial cells Nef activates the production of T-cell attracting chemokines and contributes to the development HIV infection associated brain damage. Nef also functions as an adaptor or connector protein downregulating CD4 and CCR5, the key receptor and one of the coreceptors for HIV. It also downregulates cell surface expression of a subset of class I MHC molecules which contributes to viral immune evasion. Extracellular, soluble Nef may facilitate the spread of T-cell-tropic HIV variants and mediate a switch in dominant replicating HIV strains (from macrophage-tropic to T-cell-tropic viruses) in AIDS (acquired immunodeficiency syndrome) patients. Virion-bound Nef enhances infectivity. Nef is a potential target of antiretroviral therapy and nef-deleted (attenuated) retroviruses have been considered as candidate vaccines against HIV. We suggest that nef-deleted or highly mutated defective HIV (dHIV) genomes interfere with replication of "wild type" HIV in certain long-term non-progressor individuals. This implies that introduction of artificially constructed dHIV genomes (by infusion of leukocytes carrying dHIV proviruses) into HIV infected individuals could slow disease progression and could be considered as a therapeutic possibility. PMID- 16696551 TI - Purification of a moderate thermotolerant Bacillus coagulans BTS1 lipase and its properties in a hydro-gel system. AB - An alkaline thermotolerant lipase of Bacillus coagulans BTS1 was successively purified by ammonium sulfate precipitation and DEAE anion exchange chromatography. The purified lipase immobilized in alginate beads showed an optimal activity at pH 7.5 and 55 degrees C. A pH of 5.0 or 10.0 completely quenched the activity of immobilized lipase. The alginate-bound lipase retained its activity following exposure to most of the organic solvents including amines, alkanes and alcohols. Chloride salt of Al3+, Co2+, Mg2+ and NH4+ modulated the lipase activity of alginate-immobilized enzyme. The alginate entrapped lipase showed a preferentially high activity towards p-nitrophenyl palmitate (C: 16) and activity of matrix increased following exposure to SDS. Moreover, the immobilized lipase retained more than 50% of its activity after 3rd cycle of reuse. PMID- 16696552 TI - Virulence factors and resistance mechanisms of Serratia marcescens. A short review. AB - Serratia marcescens, a Gram-negative bacillus that belongs to the family Enterobacteriaceae, is a human opportunistic pathogen bacterium that causes many diseases, such as urinary tract infections, respiratory tract infections, bacteremia, conjunctivitis, endocarditis, meningitis and wound infections. Many plasmides that confers multi-drug resistance were discovered, such as virulence factors, like cytotoxins that damage epithelial cells. The main topic of this paper presents a review about the molecular traits evolved in the pathogenic processes mediated by Serratia and its mechanism of resistance to drugs. PMID- 16696553 TI - Pulsed-field gel electrophoresis: a versatile tool for analysis of fungal genomes. A review. AB - The separation of chromosome-size DNA molecules by pulsed-field gel electrophoresis (PFGE) has become a well-established technique in recent years. Although it has very wide-ranging applications, it made a real breakthrough for fungal genome analysis. Because of the small size of fungal chromosomes, their investigation was not possible earlier. Different PFGE approaches allowed the separation of DNA molecules larger than 10 megabase pairs in size, and electrophoretic karyotypes for numerous previously genetically uncharacterized fungal species could be established. This review discusses the applicability of these electrophoretic karyotypes for the investigation of genome structure, for strain identification and for species delimitation. PMID- 16696554 TI - Early autoimmune complications after thymomectomy in a patient with interstitial lung disease. Case report. AB - Thymoma has been associated with a variety of autoimmune disorders. We report a case of myasthenia gravis and pancytopenia in a 53-year-old man with lymphoepithelial thymoma and interstitial lung disease. Preoperative examination revealed neither hematologic abnormality nor myasthenia gravis. The patient had enteritis prior to thymomectomy, sternal infection in the first month of operation, and urinary infection at the third month. About three months after thymomectomy, he required mechanical ventilation support due to myasthenia gravis related respiratory failure. One month later, a rapidly progressing pancytopenia developed. The patient died within two weeks of overwhelming septicemia unresponsive to treatment with antibiotics and steroids. The possible onset of myasthenia gravis or pancytopenia after thymomectomy should be kept in mind during follow-up. Recurrent infections in the early stages of thymomectomy may suggest a lethal onset of pancytopenia. PMID- 16696555 TI - Disruptions in maternal-infant bonding and children's respiratory systems. PMID- 16696556 TI - Theodore Xenophon Barber. PMID- 16696557 TI - Age regression: tailored versus scripted inductions. AB - The effects of tailored versus scripted hypnotic inductions were tested with the intention of shedding light on age regression phenomena. From an initial pool of 31 volunteers, 10 males and 10 females who scored 3 or better on the Stanford Hypnotic Clinical Scale, participated in this study. Participants were assigned to either scripted or tailored hypnotic induction conditions for regression to age 5. The age specific developmental task was to indicate for each of 10 abstract figure pairs, which of each pair "was upside down". Both groups showed significant focal point dependency. However, the tailored induction group showed significantly greater focal point dependency characteristic of 5-year-old children, in contrast to the scripted induction group. It appears that tailored hypnotic inductions may provide a better avenue for the ego to regulate its own degree of regression. The better match to personality style takes advantage of the naturally occurring ego-syntonic capacities of the participant, thereby facilitating greater hypnotic responsiveness. PMID- 16696558 TI - The neuroscience of observing consciousness & mirror neurons in therapeutic hypnosis. AB - Neuroscience documents the activity of "mirror neurons" in the human brain as a mechanism whereby we experience empathy and recognize the intentions of others by observing their behavior and automatically matching their brain activity. This neural basis of empathy finds support in research on dysfunctions in the mirror systems of humans with autism and fMRI research on normal subjects designed to assess intentionality, emotions, and complex cognition. Such empathy research now appears to be consistent with the historical and research literature on hypnotic induction, rapport, and many of the classical phenomena of suggestion. A preliminary outline of how mirror neurons may function as a rapport zone mediating between observing consciousness, the gene expression/protein synthesis cycle, and brain plasticity in therapeutic hypnosis and psychosomatic medicine is proposed. Brain plasticity is generalized in the theory, research, and practice of utilizing mirror neurons as an explanatory framework in developing and training new skill sets for facilitating an activity-dependent approach to creative problem solving, mind-body healing, and rehabilitation with therapeutic hypnosis. PMID- 16696559 TI - Hypnotically enhanced dreaming to achieve symptom reduction: a case study of 11 children and adolescents. AB - Theories about dreams have shaped our thinking about mind-body unity and the influence of thought on the body. In this article, the authors review the sparse literature regarding the use of hypnosis with children's dreams and nightmares, summarize how hypnotically induced dreams have been used to resolve psychological symptoms, and note five themes in the literature worthy of further investigation. Building on the value of both dreams and hypnosis for working through conflicts, the authors united mind-body medicine and hypnotically induced dreaming in a pediatric pulmonary practice. A case series is presented of 11 patients who were offered an opportunity to review their reported nightmares through hypnosis in order to uncover their potential meaning. The recurrent nightmares among these patients decreased greatly in frequency or resolved following the hypnosis enhanced dream review. Thus, we demonstrate that hypnotically induced dream review may be useful in a pediatric population. PMID- 16696560 TI - Alert hypnosis: a review and case report. AB - This review summarizes the use of hypnotic inductions while the subject is physically active, open-eyed and focused on the external environment. Research cited from several sources documents that traditional and alert inductions produce similar hypnotic susceptibility scores, but after an alert induction, subjects may report feeling more alert and in control. A case is reported of a client who was able to use such an induction to stay in alert hypnosis for an extended time, and reduce the long-standing anxiety effects of past failure. Finally, a systematic way is discussed to generate inductions that may expand the use of hypnosis to new applications. PMID- 16696562 TI - The poisons within: application of toxicity mechanisms to fundamental disease processes. PMID- 16696563 TI - Identification and quantification of a guanine-thymine intrastrand cross-link lesion induced by Cu(II)/H2O2/ascorbate. AB - Reactive oxygen species (ROS) can be induced by both endogenous and exogenous processes, and they can damage biological molecules including nucleic acids. It was shown that X- or gamma-ray irradiation of aqueous solutions of DNA, during which *OH is one of the major ROS, can lead to the formation of intrastrand cross link lesions where the neighboring nucleobases in the same DNA strand are covalently bonded. Previous 32P-postlabeling studies suggested that the intrastrand cross-link lesions may arise from Fenton reaction, which also induces the formation of *OH; the structures of the proposed intrastrand cross-link lesions, however, have not been determined. Here, we showed for the first time that the treatment of calf thymus DNA with Cu(II)/H2O2/ascorbate could lead to the formation of an intrastrand cross-link lesion, i.e., G wedge T, where the C8 of guanine is covalently bonded to the neighboring 3'-thymine through its methyl carbon. LC-MS/MS quantification results showed dose-responsive formation of G wedge T. In addition, the yield of the intrastrand cross-link was approximately 3 orders of magnitude lower than those of commonly observed single-base lesions, that is, 8-oxo-7,8-dihydro-2'-deoxyguanosine, 5-(hydroxymethyl)-2'-deoxyuridine, and 5-formyl-2'-deoxyuridine. The induction of intrastrand cross-link lesion in calf thymus DNA by Fenton reagents in vitro suggests that this type of lesion might be formed endogenously in mammalian cells. PMID- 16696564 TI - Kinetics of carboplatin-DNA binding in genomic DNA and bladder cancer cells as determined by accelerator mass spectrometry. AB - Cisplatin and carboplatin are platinum-based drugs that are widely used in cancer chemotherapy. The cytotoxicity of these drugs is mediated by platinum-DNA monoadducts and intra- and interstrand diadducts, which are formed following uptake of the drug into the nucleus of cells. The pharmacodynamics of carboplatin display fewer side effects than for cisplatin, albeit with less potency, which may be due to differences in rates of DNA adduct formation. We report the use of accelerator mass spectrometry (AMS), a sensitive detection method often used for radiocarbon quantitation, to measure both the kinetics of [14C]carboplatin-DNA adduct formation with genomic DNA and drug uptake and DNA binding in T24 human bladder cancer cells. Only carboplatin-DNA monoadducts contain radiocarbon in the platinated DNA, which allowed for calculation of kinetic rates and concentrations within the system. The percent of radiocarbon bound to salmon sperm DNA in the form of monoadducts was measured by AMS over 24 h. Knowledge of both the starting concentration of the parent carboplatin and the concentration of radiocarbon in the DNA at a variety of time points allowed calculation of the rates of Pt-DNA monoadduct formation and conversion to toxic cross-links. Importantly, the rate of carboplatin-DNA monoadduct formation was approximately 100-fold slower than that reported for the more potent cisplatin analogue, which may explain the lower toxicity of carboplatin. T24 human bladder cancer cells were incubated with a subpharmacological dose of [14C]carboplatin, and the rate of accumulation of radiocarbon in the cells and nuclear DNA was measured by AMS. The lowest concentration of radiocarbon measured was approximately 1 amol/10 microg of DNA. This sensitivity may allow the method to be used for clinical applications. PMID- 16696565 TI - A compilation of two decades of mutagenicity test results with the Ames Salmonella typhimurium and L5178Y mouse lymphoma cell mutation assays. AB - As previously reported [Cameron, T. P., Rogers-Back, A. M., Lawlor, T. E., Harbell, J. W., Seifried, H. E., and Dunkel, V. C. (1991) Gentoxicity of multifunctional acrylates in the Salmonella/mammalian-microsome assay and mouse lymphoma TK+/- assay. Environ. Mol. Mutagen. 17, 264-271], the National Cancer Institute (NCI) shares the responsibility of selecting the most significant chemicals for carcinogenicity testing by the National Toxicology Program (NTP) and has used data from Salmonella and mouse lymphoma mutagenicity assays to aid in the selection and prioritization of chemicals to be further evaluated in chronic 2 year rodent studies. In addition, a number of antineoplastic and anti AIDS drugs in preclinical evaluation were tested for the NCI's Division of Cancer Treatment Toxicology Branch. In the NCI/NTP chemical selection process, it is no longer necessary to test chemicals prior to sending them to the NTP so the NCI program has ceased performing mutagenicity tests. Some of the testing data has been made available in summary form in the Chemical Carcinogenisis Research Information System (CCRIS), which is searchable on the NLM TOXNET system. The limitations in using this source are that only summary results are available and many negative test results are not included. A summary table that presents the results for each compound is provided in the Appendix with raw data provided in the Supporting Information. The Appendix table contains the compound name, CAS number, and a summary of the data from the Ames test and the mouse lymphoma assay. PMID- 16696566 TI - Cross-linking of the human DNA repair protein O6-alkylguanine DNA alkyltransferase to DNA in the presence of 1,2,3,4-diepoxybutane. AB - 1,2,3,4-Diepoxybutane (DEB) is a key carcinogenic metabolite of the important industrial chemical 1,3-butadiene. DEB is a bifunctional alkylating agent capable of reacting with DNA and proteins. Initial DNA alkylation by DEB produces N7-(2' hydroxy-3',4'-epoxybut-1'-yl)-guanine monoadducts, which can react with another nucleophilic site to form cross-linked adducts. A recent report revealed a strong correlation between cellular expression of the DNA repair protein O6-alkylguanine DNA alkyltransferase (AGT) and the cytotoxic and mutagenic activity of DEB, suggesting that DEB induces AGT-DNA cross-links (Valadez, J. G., et al. (2004) Activation of bis-electrophiles to mutagenic conjugates by human O6-alkylguanine DNA alkyltransferase. Chem. Res. Toxicol. 17, 972-982). The purpose of our study was to analyze the formation and structures of DEB-induced AGT-DNA conjugates and to identify specific amino acid residues within the protein involved in cross linking. DNA-protein cross-link formation was detected by SDS-PAGE when 32P labeled double-stranded oligodeoxynucleotides were exposed to DEB in the presence of either wild-type hAGT or a C145A hAGT mutant. Capillary HPLC-electrospray ionization mass spectrometry (ESI-MS) analysis of hAGT that had been treated with N7-(2'-hydroxy-3',4'-epoxybut-1'-yl)-deoxyguanosine (dG monoepoxide) revealed the ability of the protein to form either one or two butanediol-dG cross-links, corresponding to mass shifts of +353 and +706 Da, respectively. HPLC-ESI+ -MS/MS sequencing of the tryptic peptides obtained from dG monoepoxide-treated protein indicated that the two cross-linking sites were the alkyl acceptor site, Cys145, and a neighboring active site residue, Cys150. The same two amino acid residues of hAGT became covalently cross-linked to DNA following DEB treatment. Modification of Cys145 was further confirmed by HPLC-ESI+ -MS/MS analysis of dG monoepoxide-treated synthetic peptide GNPVPILIPCHR which represents the active site tryptic fragment of hAGT (C = Cys145). The replacement of the catalytic cysteine residue with alanine in the C145A hAGT mutant abolished DEB-induced cross-linking at this site, while the formation of conjugates via neighboring Cys150 was retained. The exact chemical structure of the cross-linked lesion was established as 1-(S-cysteinyl)-4-(guan-7-yl)-2,3-butanediol by HPLC-ESI+ -MS/MS analysis of the amino acids resulting from the total digestion of modified proteins analyzed in parallel with an authentic standard. AGT-DNA cross-linking is a likely mechanism of DEB-mediated cytotoxicity in cells expressing this important repair protein. PMID- 16696568 TI - The behavior of mixtures of paralytic shellfish toxins in competitive binding assays. AB - Organisms that contain paralytic shellfish toxins (PSTs) may contain many members of this toxin family. PSTs block voltage-gated sodium channels (Na channel) and elicit neurotoxicity. Animals, including humans, may encounter PST mixtures via consumption of tainted seafood, contaminated water, or the microalgae that produce the toxins. PST binding by the Na channel as well as other proteins such as antibodies and saxiphilin have been used to develop biomolecular assays for PSTs. An equation that predicts the combined effects of binary and ternary PST mixtures has been experimentally validated for two unrelated STX-binding proteins, the rat brain Na channel and a saxiphilin from the xanthid crab Liomera tristis. It was found that the most potent toxin or toxins in any mixture profoundly affect the cumulative potency of the mixture, overwhelming weaker toxins with the transition from strong to weak toxicity and changing in a curvilinear manner. Less active PSTs must be several orders of magnitude more concentrated than stronger toxins for the mixture to reflect their potency. This behavior is important in understanding how toxin mixtures may act at the Na channel receptor via which PSTs exert their neurotoxicity and that the presence of weaker toxins does not dilute the effect of stronger toxins in a linear fashion. This strong dominance of a mixture by the most potent toxins also has implications for measurement of toxic test samples and for standards that may contain low levels of highly potent bioactive impurities. This equation has been extended to mixtures of PSTs containing more than three toxins and may be applicable to other natural contaminants and any competitive binding assays used to detect their presence and measure their concentration. PMID- 16696567 TI - Ebselen induced C6 glioma cell death in oxygen and glucose deprivation. AB - Studies have shown that ebselen is an antiinflammatory and antioxidative agent. Its protective effect has been investigated in oxidative stress related diseases such as cerebral ischemia in recent years. However, experimental evidence also shows that ebselen causes cell death in several different cell types. Whether ebselen will have a beneficial or detrimental effect on cells under ischemic condition is not known. Herein, we studied the effect of ebselen on C6 glioma cells under oxygen and glucose deprivation (OGD), an in vitro ischemic model. We found that ebselen significantly enhanced cell death after 3 h of OGD as observed by lactase dehydrogenase (LDH) release and cellular morphological changes. Further studies revealed that depletion of cellular glutathione level by the combined action of ebselen and OGD played a role in enhanced cell death as demonstrated by the following evidence: (1) cellular GSH was significantly depleted by the combined effort of ebselen and OGD, compared to that of ebselen or OGD insult alone; (2) exogenous addition of N-acetyl cysteine completely diminished the cell damage induced by ebselen and OGD; (3) supplement of glucose, which provides cellular reducing agents and thus maintains cellular GSH level, to the OGD medium diminished C6 cell damage induced by ebselen. We conclude that depleting cellular glutathione plays an important role in ebselen-induced cell death with OGD. Our results suggest that ebselen can have a beneficial or toxic effect, depending on the availability of GSH. PMID- 16696569 TI - Effect of heparin on viologen-stimulated enzymatic NADH depletion. AB - Paraquat and diquat undergo redox cycling mediated by xanthine oxidase in the NADH-dependent manner. In these processes, the rates of NADH oxidation and superoxide formation are increased almost 10-fold. The addition of heparin can substantially inhibit these processes. A protective role of heparin against oxygen radicals formation can be rationalized in terms of its ability to bind paraquat or diquat. The binding process has been investigated by means of the pulse radiolysis technique. Biological consequences of the binding processes are discussed. PMID- 16696570 TI - Quantitation of pyridyloxobutyl DNA adducts of tobacco-specific nitrosamines in rat tissue DNA by high-performance liquid chromatography-electrospray ionization tandem mass spectrometry. AB - The tobacco-specific nitrosamines N'-nitrosonornicotine (NNN, 1) and 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK, 2) are potent carcinogens in rodents. Bioactivation of NNN and NNK by cytochrome P450 enzymes generates a pyridyloxobutylating agent 6, which alkylates DNA to produce pyridyloxobutyl (POB)-DNA adducts. POB-DNA adduct formation plays a critical role in NNN and NNK carcinogenicity in rodents. To further investigate the significance of this pathway, we developed a high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS) method for quantitative analysis of four POB-DNA adducts with known structures. The corresponding deuterated analogues were synthesized and used as internal standards. DNA samples, spiked with internal standards, were subjected to neutral thermal hydrolysis followed by enzymatic hydrolysis. The hydrolysates were partially purified by solid phase extraction prior to HPLC-ESI-MS/MS analysis. The method was accurate and precise. Excellent sensitivity was achieved, especially for O2 [4-(3-pyridyl)-4-oxobut-1-yl]thymidine (O2-POB-dThd, 11) with a detection limit of 100 amol per mg DNA. DNA samples treated with different concentrations of 4 (acetoxymethylnitrosamino)-1-(3-pyridyl)-1-butanone (NNKOAc, 3) were subjected to HPLC-ESI-MS/MS analysis. 7-[4-(3-Pyridyl)-4-oxobut-1-yl]guanine (7-POB-Gua, 12) was the most abundant adduct, followed by O6-[4-(3-pyridyl)-4-oxobut-1-yl]-2' deoxyguanosine (O6-POB-dGuo, 8), O2-POB-dThd, and O2-[4-(3-pyridyl)-4-oxobut-1 yl]cytosine (O2-POB-Cyt, 13). Lung and liver DNA isolated from NNK-treated rats were analyzed. Consistent with the in vitro data, 7-POB-Gua was the major POB-DNA adduct formed in vivo. However, levels of O6-POB-dGuo were the lowest of the four adducts analyzed, suggesting efficient repair of this adduct in vivo. In contrast to the other three adducts, O6-POB-dGuo was more abundant in lung than in liver. O2-POB-dThd appeared to be poorly repaired in vivo, and its levels were comparable to those of 7-POB-Gua. The results of this study provide a sensitive HPLC-ESI-MS/MS method for comprehensive quantitation of four POB-DNA adducts, support an important role of O6-POB-dGuo in NNK lung tumorigenicity in rats, and suggest that O2-POB-dThd may be a useful tobacco-specific DNA biomarker for future tobacco carcinogenesis studies. PMID- 16696571 TI - Bioactivation of carboxylic acid compounds by UDP-Glucuronosyltransferases to DNA damaging intermediates: role of glycoxidation and oxidative stress in genotoxicity. AB - Nonenzymatic modification of proteins by acyl glucuronides is well documented; however, little is known about their potential to damage DNA. We have previously reported that clofibric acid undergoes glucuronidation-dependent bioactivation to DNA-damaging species in cultured mouse hepatocytes. The aim of this study was to investigate the mechanisms underlying such DNA damage, and to screen chemically diverse carboxylic acid drugs for their DNA-damaging potential in glucuronidation proficient murine hepatocytes. Cells were incubated with each aglycone for 18 h, followed by assessment of compound cytotoxicity using the MTT assay and evaluation of DNA damage using the Comet assay. Relative cytotoxic potencies were ketoprofen > diclofenac, benoxaprofen, nafenopin >> gemfibrozil, probenecid > bezafibrate > clofibric acid. At a noncytotoxic (0.1 mM) concentration, only benoxaprofen, nafenopin, clofibric acid, and probenecid significantly increased Comet moments (P < 0.05 Kruskal-Wallis). Clofibric acid and probenecid exhibited the greatest DNA-damaging potency, producing significant DNA damage at 0.01 mM concentrations. The two drugs produced maximal increases in Comet moment of 4.51 x and 2.57 x control, respectively. The glucuronidation inhibitor borneol (1 mM) abolished the induction of DNA damage by 0.5 mM concentrations of clofibric acid and probenecid. In an in vitro cell-free system, clofibric acid glucuronide was 10 x more potent than glucuronic acid in causing DNA strand-nicking, although both compounds showed similar rates of autoxidation to generate hydroxyl radicals. In cultured hepatocytes, the glycation inhibitor, aminoguanidine, and the iron chelator, desferrioxamine mesylate, inhibited DNA damage by clofibric acid, whereas the free radical scavengers Trolox and butylated hydroxytoluene, and the superoxide dismutase mimetic bis-3,5-diisopropylsalicylate had no effect. In conclusion, clinically relevant concentrations of two structurally unrelated carboxylic acids, probenecid and clofibric acid, induced DNA damage in isolated hepatocytes via glucuronidation- dependent pathways. These findings suggest acyl glucuronides are able to access and damage nuclear DNA via iron-catalyzed glycation/glycoxidative processes. PMID- 16696572 TI - Identification of a novel hemoglobin adduct in Sprague Dawley rats exposed to atrazine. AB - Atrazine (2-chloro-4-[ethylamino]-6-[isopropylamino]-1,3,5-triazine) is one of the most commonly used herbicides in North America and is frequently detected in ground and surface waters. This research investigated possible covalent modifications of hemoglobin following in vivo exposures to atrazine in Sprague Dawley (SD) rats and in vitro incubations with diaminochlorotriazine. SD rats were exposed to 0, 10, 30, 100, and 300 (mg atrazine/kg)/day for 3 days via oral gavages, and blood was drawn at 0 h, 24 h, 72 h, 20 days, 1 month, and 2 months for globin analysis. Globin was purified from red blood cells, separated with high-performance liquid chromatography, and analyzed with matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS). An additional beta globin peak was seen in exposed animals during the HPLC and MALDI TOF MS analysis with a mass 110 Da greater than the normal beta subunits. Tryptic digests of this beta peak contained a peptide of 1449.9 m/z that corresponded to a modified peptide of amino acids 121-132. Mass spectrometry sequencing of this peptide indicated a 110 Da addition to Cys-125 of the major beta globin chain, which corresponds to a nucleophilic substitution reaction with a diaminochlorotriazine. In vitro incubations of SD globin and diaminochlorotriazine also resulted in a peptide of 1449.6 m/z that was identical in sequence to the modified peptide seen in the in vivo digest, confirming the nucleophilic substitution mechanism of adduct formation. Exposures of SD rats to atrazine results in formation of an adduct that is easily detected and provides an analytical model for detection of triazine adducts in other macromolecules with sulfhydryl functional groups. PMID- 16696573 TI - Kinetics of acetaminophen glucuronidation by UDP-glucuronosyltransferases 1A1, 1A6, 1A9 and 2B15. Potential implications in acetaminophen-induced hepatotoxicity. AB - The importance of uridine 5'-diphosphate-glucuronosyltranferases (UGT) 2B15 and other UGT enzymes (1A1, 1A6, and 1A9) in glucuronidating acetaminophen (APAP) is demonstrated. The kinetics and contributions of various UGTs in glucuronidating APAP are presented using clinically and toxicologically relevant concentrations of the substrate. UGT 1A9 and UGT 2B15 contribute significantly toward glucuronidating APAP when incubations were conducted in either phosphate or Tris HCl buffers at 0.1 and 1.0 mM substrate concentrations. At 10 mM APAP, UGT 1A9 is a significant enzyme responsible for metabolizing APAP in either one of the buffers. UGT 1A1 is the next most important enzyme in glucuronidating APAP at this high substrate concentration. The contribution of UGT 1A6 at 10 mM APAP concentration became obscured by similar relative activities exhibited by UGTs 1A7, 1A8, and 2B7. These observations may reflect the differences in kinetic parameters for APAP glucuronidation by the individual UGTs. UGT 1A1 demonstrated Hill kinetics while UGT 1A9 displayed Michaelis-Menten kinetics. Substrate inhibition kinetics is observed with UGT 1A6, UGT 2B15, and human liver microsomes. The substrate inhibition is confirmed by employing stable isotope labeled APAP as the substrate, while APAP glucuronide is used to test for inhibition of d4-APAP glucuronide. The in vitro hepatotoxicity caused by APAP in combination with phenobarbital or phenytoin is demonstrated in this study. The inhibition of APAP glucuronidation by phenobarbital leads to an increase in APAP mediated toxicity in human hepatocytes. The toxicity to hepatocytes was further increased by coadministering APAP with phenytoin and phenobarbital. This synergistic increase in toxicity is postulated to be due to inhibition of UGTs (1A6, 1A9, and 2B15) responsible for detoxifying APAP through the glucuronidation pathway. PMID- 16696574 TI - Detection and quantification of endogenous cyclic DNA adducts derived from trans 4-hydroxy-2-nonenal in human brain tissue by isotope dilution capillary liquid chromatography nanoelectrospray tandem mass spectrometry. AB - A sensitive and selective capillary liquid chromatography nanoelectrospray isotope dilution mass spectrometric method was developed to identify and quantify the endogenous cyclic DNA adducts derived from trans-4-hydroxy-2-nonenal with 2' deoxyguanosine (HNE-dG) in human brain tissues. Authentic and 13C and 15N stable isotope-labeled HNE-dG were synthesized to serve as standards. The in vitro HNE modified calf-thymus DNA as well as the DNA samples isolated from human brain tissues of normal and Alzheimer's disease subjects were enzymatically digested to nucleosides in vitro with the presence of internal standard (HNE-dG-13C10, 15N5). The enzymatic digests were cleaned up by solid phase extraction. Only 1-2 microg of DNA digests was loaded on a laboratory-constructed reversed phase capillary chromatography column, and the HNE-dG adducts were separated from intact nucleosides and quantified by a high capacity ion trap mass spectrometer in the MS/MS mode. This method was able to quantify an adduct level of approximately 40 lesions/10(9) normal DNA nucleosides. The detected level of HNE-dG adducts in hippocampus/parahippocampal gyrus and inferior parietal regions of postmortem brains from AD subjects were 556 +/- 379 and 238 +/- 72 adducts per 10(9) normal nucleosides, respectively. These results were consistent with the 32P postlabeling results, which detected 400-600 adducts per 10(9) normal nucleotides in the hippocampus. PMID- 16696575 TI - Polycyclic aromatic hydrocarbon (PAH) o-quinones produced by the aldo-keto reductases (AKRs) generate abasic sites, oxidized pyrimidines, and 8-oxo-dGuo via reactive oxygen species. AB - Reactive and redox-active polycyclic aromatic hydrocarbon (PAH) o-quinones produced by Aldo-Keto Reductases (AKRs) have the potential to cause depurinating adducts leading to the formation of abasic sites and oxidative base lesions. The aldehyde reactive probe (ARP) was used to detect these lesions in calf thymus DNA treated with three PAH o-quinones (BP-7,8-dione, 7,12-DMBA-3,4-dione, and BA-3,4 dione) in the absence and presence of redox-cycling conditions. In the absence of redox-cycling, a modest amount of abasic sites were detected indicating the formation of a low level of covalent o-quinone depurinating adducts (>3.2 x 10(6) dNs). In the presence of NADPH and CuCl2, the three PAH o-quinones increased the formation of abasic sites due to ROS-derived lesions destabilizing the N glycosidic bond. The predominant source of AP sites, however, was revealed by coupling the assay with human 8-oxoguanine glycosylase (hOGG1) treatment, showing that 8-oxo-dGuo was the major lesion caused by PAH o-quinones. The levels of 8 oxo-dGuo formation were independently validated by HPLC-ECD analysis. Apyrimidinic sites were also revealed by coupling the assay with Escherichia coli (Endo III) treatment showing that oxidized pyrimidines were formed, but to a lesser extent. Different mechanisms were responsible for the formation of the oxidative lesions depending on whether Cu(II) or Fe(III) was used in the redox cycling conditions. In the presence of Cu(II)-mediated PAH o-quinone redox cycling, catalase completely suppressed the formation of the lesions, but mannitol and sodium benzoate were without effect. By contrast, sodium azide, which acts as a *OH and 1O2 scavenger, inhibited the formation of all oxidative lesions, suggesting that the ROS responsible was 1O2. However, in the presence of Fe(III)-mediated PAH o-quinone redox-cycling, the *OH radical scavengers and sodium azide consistently attenuated their formation, indicating that the ROS responsible was *OH. PMID- 16696576 TI - Role of melatonin in the pathophysiology of migraine: implications for treatment. AB - The pineal gland and its secretory product, melatonin, have been implicated in the pathophysiology of migraine, as well as some of its comorbid disorders. Supporting this view, many migraineurs are susceptible to various environmental triggers that influence the secretion of melatonin from the pineal gland, and many prodromal symptoms are probably generated in the hypothalamus, a brain region that provides input into the pineal gland to modulate the secretion of melatonin. In addition, studies have shown abnormalities in melatonin secretion in patients who experience migraine and an improvement in migraine following administration of melatonin. However, the dysfunction in melatonin secretion in migraineurs may simply be a marker of hypothalamic dysfunction or neuronal hyperexcitability, leading to migraine susceptibility. It is also possible that abnormal melatonin secretion leads to decreased inhibitory neurotransmitter activity, decreased inhibition of the release of calcitonin gene-related peptide (CGRP) from activation of the trigeminal system, or less analgesia. Whatever its role in the pathogenesis of migraine, melatonin may prove to be a useful therapy. Future studies are necessary to further elucidate whether melatonin is a well tolerated, beneficial therapy, and to determine the optimal dose and formulation of melatonin for use in migraine therapy. PMID- 16696578 TI - Management of epilepsy in women of childbearing age: practical recommendations. AB - Women with epilepsy should not be discouraged from becoming pregnant as the likelihood of having a healthy baby is very high. However, in such women, early and individualised counselling about pregnancy and contraception is essential. Ideally, pregnancies should be planned, folic acid (5 mg/day) given and antiepileptic drug (AED) treatment optimised well before conception to ensure that the lowest dosage that controls seizures is administered. When initiating AEDs in a woman of childbearing age, the most appropriate drug for the seizure type and syndrome should be chosen, although it is preferable to avoid valproate, because of a possible elevated risk of fetal malformations, when equi-effective agents are available for a given syndrome. In women who become pregnant while taking AEDs, fetal monitoring should include high-resolution ultrasonography before week 20 and measurement of serum alpha-fetoprotein levels. Amniocentesis is not routinely indicated. The measurement of blood concentrations of AEDs can be useful to ensure that the lowest possible maintenance dosage is being used, especially for those drugs whose pharmacokinetics are likely to change during pregnancy. Breastfeeding should be encouraged whatever the treatment administered. PMID- 16696577 TI - Inhibition of gamma-secretase as a therapeutic intervention for Alzheimer's disease: prospects, limitations and strategies. AB - Genetic and experimental evidence points to amyloid-beta (Abeta) peptide as the culprit in Alzheimer's disease pathogenesis. This protein fragment abnormally accumulates in the brain cortex and hippocampus of patients with Alzheimer's disease, and self-aggregates to form toxic oligomers causing neurodegeneration.Abeta is heterogeneous and produced from a precursor protein (amyloid precursor protein [APP]) by two sequential proteolytic cleavages that involve beta- and gamma-secretases. This latter enzyme represents a potentially attractive drug target since it dictates the solubility of the generated Abeta fragment by creating peptides of various lengths, namely Abeta(40) and Abeta(42), the longest being the most aggregating. gamma-Secretase comprises a molecular complex of four integral membrane proteins - presenilin, nicastrin, APH-1 and PEN 2 - and its molecular mechanism remains under extensive scrutiny. The ratio of Abeta(42) over Abeta(40) is increased by familial Alzheimer's disease mutations occurring in the presenilin genes or in APP, near the gamma-secretase cleavage site. Potent gamma-secretase inhibitors have been identified by screening drug libraries or by designing aspartyl protease transition-state analogues based on the APP substrate cleavage site. Most of these compounds are not specific for gamma-secretase cleavage of APP, and equally inhibit the processing of other gamma-secretase substrates, such as Notch and a subset of cell-surface receptors and proteins involved in embryonic development, haematopoiesis, cell adhesion and cell/cell contacts. Therefore, current research aims at finding compounds that show selectivity for APP cleavage, and particularly that inhibit the formation of the aggregating form, Abeta(42). Compounds that target the substrate docking site rather than the enzyme active site are also being investigated as an alternative strategy. The finding that some NSAID analogues preferentially inhibit the formation of Abeta(42) over Abeta(40) and do not affect Notch processing has opened a new therapeutic window. The progress in design of selective inhibitors as well as recent results obtained in animal studies prove that gamma-secretase remains among the best targets for the therapeutic control of amyloid build-up in Alzheimer's disease. The full understanding of gamma-secretase regulation may yet uncover new therapeutic leads. PMID- 16696580 TI - Cardiovascular effects and risk of syncope related to donepezil in patients with Alzheimer's disease. AB - BACKGROUND: When otherwise unexplained, syncope in patients with Alzheimer's disease may be attributed to bradycardia caused by cholinesterase inhibitors. We studied prospectively the clinical events and cardiovascular changes occurring during treatment with donepezil in patients with Alzheimer's disease. METHODS: Consecutive patients presenting with mild-to-moderate Alzheimer's disease were included in the study. Their clinical characteristics, blood pressure, heart rate and electrocardiogram were recorded before (baseline) and during treatment with donepezil. The drug was administered at a dosage of 5 mg/day for 1 month and 10 mg/day for the following 7 months, as tolerated. We compared the baseline observations with those made at 1, 2 and 8 months of donepezil treatment. We also examined the effects of negatively chronotropic or dromotropic drugs concomitantly administered with donepezil. RESULTS: Thirty patients were included in the study, of whom 43% were taking negatively chronotropic or dromotropic drugs. The first month of therapy (donepezil 5 mg/day) was completed by 26 patients. During the 7-month high-dosage phase (10 mg/day), four patients dropped out of the study; thus, 22 patients completed the full 8 months of the study. The mean heart rate was 66 +/- 8 beats/min at baseline in the overall study population. This decreased significantly to 62 +/- 9, 61 +/- 7 and 62 +/- 8 beats/min at the 1, 2 and 8 month timepoints, respectively (all p = 0.002 vs baseline). Among patients not receiving negatively chronotropic or dromotropic drugs, heart rate decreased significantly over the course of the study (from 67 +/- 8 beats/min at baseline to 62 +/- 8 beats/min at 1 month, 62 +/- 7 beats/min at 2 months and 62 +/- 8 beats/min at 8 months [all p = 0.005 vs baseline]). There was no significant change in heart rate in patients who were receiving negatively chronotropic or dromotropic drugs. The PR interval increased over the course of the study in all patient groups, but these changes were only statistically significant in the group of patients who were not taking negatively chronotropic or dromotropic drugs (155 +/- 23ms at baseline vs 158 +/- 21, 160 +/ 22 and 163 +/- 24ms at the 1, 2 and 8 month timepoints; all p = 0.02 vs baseline). One patient developed syncope due to orthostatic hypotension; there were no cases of bradycardia-induced syncope. Gastrointestinal manifestations were reported in ten of the study patients. Abdominal pain and vomiting were the reasons for study termination in five of the eight patients who did not complete the trial. CONCLUSION: A donepezil-induced decrease in heart rate and increase in PR interval were observed only in patients with Alzheimer's disease who were not treated with negatively chronotropic or dromotropic drugs. These changes were not associated with bradycardia-induced syncope. PMID- 16696579 TI - Mechanism of action of atypical antipsychotic drugs and the neurobiology of schizophrenia. AB - Atypical antipsychotics have greatly enhanced the treatment of schizophrenia. The mechanisms underlying the effectiveness and adverse effects of these drugs are, to date, not sufficiently explained. This article summarises the hypothetical mechanisms of action of atypical antipsychotics with respect to the neurobiology of schizophrenia.When considering treatment models for schizophrenia, the role of dopamine receptor blockade and modulation remains dominant. The optimal occupancy of dopamine D(2) receptors seems to be crucial to balancing efficacy and adverse effects - transient D(2) receptor antagonism (such as that attained with, for example, quetiapine and clozapine) is sufficient to obtain an antipsychotic effect, while permanent D(2) receptor antagonism (as is caused by conventional antipsychotics) increases the risk of adverse effects such as extrapyramidal symptoms. Partial D(2) receptor agonism (induced by aripiprazole) offers the possibility of maintaining optimal blockade and function of D(2) receptors. Balancing presynaptic and postsynaptic D(2) receptor antagonism (e.g. induced by amisulpride) is another mechanism that can, through increased release of endogenous dopamine in the striatum, protect against excessive blockade of D(2) receptors. Serotonergic modulation is associated with a beneficial increase in striatal dopamine release. Effects on the negative and cognitive symptoms of schizophrenia relate to dopamine release in the prefrontal cortex; this can be modulated by combined D(2) and serotonin 5-HT(2A) receptor antagonism (e.g. by olanzapine and risperidone), partial D(2) receptor antagonism or the preferential blockade of inhibitory dopamine autoreceptors. In the context of the neurodevelopmental disconnection hypothesis of schizophrenia, atypical antipsychotics (in contrast to conventional antipsychotics) induce neuronal plasticity and synaptic remodelling, not only in the striatum but also in other brain areas such as the prefrontal cortex and hippocampus. This mechanism may normalise glutamatergic dysfunction and structural abnormalities and affect the core pathophysiological substrates for schizophrenia. PMID- 16696581 TI - Zolpidem extended-release. AB - Zolpidem extended-release, or controlled-release (CR), is a new formulation of zolpidem, a nonbenzodiazepine hypnotic. It is indicated in the US for the treatment of insomnia, characterised by difficulties with sleep onset and/or sleep maintenance. Zolpidem CR is a dual-layered tablet; one layer releases zolpidem immediately and a second layer provides a slower release of additional zolpidem for maintenance of plasma zolpidem concentrations. Efficacy of zolpidem CR was assessed in two 3-week, randomised, double-blind, placebo-controlled, phase III polysomnography trials in younger adult (aged 18-64 years) or elderly (aged > or =65 years) patients with primary insomnia. Patients received nightly zolpidem CR (12.5mg in younger adult and 6.25mg in elderly patients). Efficacy was assessed objectively on nights 1, 2, 15 and 16. Patients who received zolpidem CR had significantly improved objective latency to persistent sleep, wake time after sleep onset and sleep efficiency on assessment nights compared with placebo recipients. In subjective assessments of sleep quality on day 2 and nights 15 and 22, significantly more zolpidem CR than placebo recipients gave favourable responses on a Patient Global Impression scale in the study in younger adult patients. In the other study, significantly more elderly patients in the zolpidem CR group rated their sleep as improved compared with the placebo group. Zolpidem CR was generally well tolerated and appears to have a tolerability profile similar to that of the original formulation of zolpidem. PMID- 16696584 TI - Spotlight on quetiapine in acute mania and depression associated with bipolar disorder. AB - Quetiapine (Seroquel), an atypical antipsychotic with established efficacy in the treatment of schizophrenia, shows efficacy in the treatment of acute mania and depression associated with bipolar disorder.Quetiapine, either as monotherapy or in combination with lithium or divalproex sodium (valproate semisodium), is generally well tolerated and effective in reducing manic symptoms in adult and adolescent patients with acute bipolar mania, and is approved for use in adults for this indication. As monotherapy, the drug is also effective in reducing depressive symptoms in patients with bipolar depression. It is associated with a low incidence of extrapyramidal symptom (EPS)-related adverse events and low EPS ratings in bipolar disorder. Quetiapine thus shows potential in the treatment of bipolar depression, and represents a useful agent for the treatment of acute bipolar mania. PMID- 16696585 TI - The role of antihistamines in asthma management. AB - Histamine is an important mediator in airway inflammation. It is elevated in the airways of asthmatic patients and is responsible for many of the pathophysiological features in asthma. Antihistamines block the actions of histamine and also have effects on inflammation which is independent of histamine H(1)-receptor antagonism. Antihistamines have been shown to have bronchodilatory effects, effects on allergen-, exercise-, and adenosine-monophosphate-challenge testing, and also to prevent allergen-induced nonspecific airways hyperresponsiveness. Clinical studies have shown mixed results, and some studies have reported beneficial effects of azelastine, cetirizine, desloratadine, and fexofenadine on asthma symptoms or physiological measures in patients with asthma. The combination of an antihistamine and a leukotriene receptor antagonist has been shown to have additive effects in certain studies. Antihistamines have also been shown to delay or prevent the development of asthma in a subgroup of atopic children. These data suggest that antihistamines may have beneficial effects in the management of asthma. PMID- 16696586 TI - Chemokine receptors : therapeutic potential in asthma. AB - Leukocyte infiltration of the lung is a characteristic feature of allergic asthma and it is thought that these cells are selectively recruited by chemokines. Extensive research has confirmed that chemokine receptors are expressed on the main cell types involved in asthma, including eosinophils, T helper type 2 cells, mast cells and even neutrophils. Moreover, animal experiments have outlined a functional role for these receptors and their ligands. Chemokines signal via seven-transmembrane spanning G-protein coupled receptors, which are favored targets of the pharmaceutical industry due to the possibility of designing small molecule inhibitors. In fact, this family represents the first group of cytokines where small-molecule inhibitors have been designed. However, the search for efficient antagonists of chemokine/chemokine receptors has not been easy; a particular feature of the chemokine system is the number of molecules with overlapping functions and binding specificities, as well as the difficulty in reconciling the in vivo biologic functional validation of chemokines in rodent models with the development of antagonists which bind the human receptor, because of the lack of species cross-reactivity. The chemokines and their receptors that are active during allergic reactions are reviewed. Possible points of interaction that may be a target for development of new therapies, as well as the progress to date in developing inhibitors of key chemokine receptors for asthma therapy, are also discussed. PMID- 16696587 TI - Hypersensitivity pneumonitis : a broader perspective. AB - Hypersensitivity pneumonitis (HP) represents a group of lung disorders caused by the inhalation of a wide variety of organic particles by susceptible individuals. HP occurs mainly in nonsmokers, but smoking may promote an insidious and chronic disease. The prevalence of HP is difficult to estimate accurately since several antigens can produce the disease, but the range spans infancy to old age. Regardless of the causative antigen or its environmental setting, the clinical manifestations are essentially the same. Three different clinical presentations have been recognized: acute, subacute, and chronic. In the acute form, patients show flu-like symptomatology, followed by dyspnea and dry cough. Symptoms subside a few hours or days later. The subacute and chronic forms result from recurrent low-level antigen exposure and are characterized by progressive dyspnea and dry cough. Other constitutional symptoms such as fatigue, anorexia, and weight loss can be apparent. Fever may occur in the subacute form. Importantly, chronic HP may evolve insidiously or may result from repeated acute/subacute episodes. Recurrent acute, subacute, and chronic HP may progress to irreversible lung fibrosis or provoke emphysematous changes.HP can be difficult to identify, and precise diagnosis requires a history of exposure and a constellation of clinical, imaging, laboratory, bronchoalveolar lavage and pathologic findings. General laboratory tests show an increase of acute phase reactants. Specific precipitating antibodies, when present, are evidence of antigen exposure, and are a hallmark for diagnosis. Chest radiograph usually reveals widespread ground glass attenuation, and nodular or reticulonodular shadowing. High-resolution CT features include diffuse or patchy ground-glass opacities with small poorly defined nodules and air trapping. Pulmonary function tests are characterized by a predominantly restrictive ventilatory defect with loss of lung volume and hypoxemia at rest that worsens with exercise. Bronchoalveolar lavage reveals a significant increase in lymphocytes, mostly over 40%. In the acute form there is also an increase in neutrophils. Antigen-induced lymphocyte proliferation, and environmental or laboratory-controlled inhalation challenge, may be used for diagnostic purposes and can help to establish a diagnosis of insidious forms of HP. In subacute or chronic cases, lung biopsy may be necessary. Typical findings include bronchiolitis, lymphocytic alveolitis, and loosely formed granulomas, although occasionally other morphologic patterns such as nonspecific interstitial pneumonia may exist. Treatment focuses on avoiding further exposure to the offending antigen(s). Corticosteroids are recommended in subacute and chronic forms. The usual regimen consists of initial high doses of systemic corticosteroid (e.g. prednisone 0.5-1.0 mg/kg/day), followed by gradual tapering. PMID- 16696588 TI - Taxanes in the treatment of non-small cell lung cancer. AB - Paclitaxel and docetaxel, drugs that bind tightly to beta-tubulin and disrupt microtubule dynamics, are widely used in the treatment of non-small cell lung cancer (NSCLC), the most common cause of cancer death in men and women living in the US. These well tolerated drugs, alone or in combination with another cytotoxic agent, have been shown to increase the survival of patients with metastatic disease or malignant effusions. Both paclitaxel and docetaxel can be combined with concurrent chest irradiation for patients with locally advanced NSCLC. The combination of carboplatin and paclitaxel, when given postoperatively to patients with stage IB NSCLC, improved survival compared with surgery alone, with little toxicity. Taxane combinations are undergoing study as adjuvant therapy for patients with other stages of operable disease. Except for a recent trial with bevacizumab, efforts to improve the efficacy of taxane/platinum combinations in patients with advanced disease by adding a third 'targeted' drug have thus far been unsuccessful. PMID- 16696589 TI - The organizing pneumonias : a critical review of current concepts and treatment. AB - In this comprehensive review, two very closely related interstitial pneumonias are discussed: the cryptogenic form of organizing pneumonia (COP); and secondary forms of organizing pneumonia (OP), which occur in association with identifiable medical conditions. Some newer and lesser known of these associated conditions are described, most importantly post-radiation OP.Rapidly progressive, corticosteroid-resistant and poor prognostic forms of OP have been described. These types purportedly occur more frequently in secondary OP. However, OPs frequently coexist with other interstitial pneumonias, especially when associated with connective tissue diseases. Therefore, tissue sampling error or an incorrect morphologic diagnosis can be the basis for the occurrence of clinically aggressive OPs. By using the 2002 American Thoracic Society/European Respiratory Society diagnostic criteria, some pre-2002 cases reported as OP would be re classified today.Although COP is considered to have a good prognosis and to be corticosteroid responsive, approximately 70% of patients, treated with corticosteroids, relapse even during initial treatment. Multiple and late relapses occur in about one-third of the patients. We performed a meta-analysis of second-line treatment options for corticosteroid-refractory forms of OP. Three alternative nonsteroid agents - cyclophosphamide, azathioprine, and cyclosporin - have been used in combination with corticosteroids. On careful review, in a number of cases reported as secondary OP, other histologic interstitial patterns besides OP were described. The need for second-line therapy in these patients might have been dictated by the non-organizing pneumonic component. Most of the scant number of reports come from outside the US. World experience with these is limited, but good clinical outcomes have been noted, even in patients with interstitial patterns in addition to OP.The initiation of the OP tissue response in the bronchiolar and sub-bronchiolar location may be due to the presence of bronchiolar-associated lymphoid tissue found at the bifurcations of the bronchioles. Inhaled antigens stimulate granulocyte colony stimulating factor mediated airway inflammation, followed later by CD44-mediated clearance. Repair requires intrabronchiolar formation of granulation tissue and a favorable ratio of matrix metalloproteinase to tissue inhibitors of metalloproteinase (MMP : TIMP) within the stroma. This reparative milieu allows extracellular matrix degradation and re-synthesis to occur. MMP-expressing fibroblasts then phagocytose the collagen fibrils and microfibrils produced earlier in repair, reversing the initial fibrosis. PMID- 16696590 TI - The potential role of phosphodiesterase inhibitors in the management of asthma. AB - Asthma is a chronic inflammatory condition characterised by reversible airflow obstruction and airway hyperreactivity. The course of the illness may be punctuated by exacerbations resulting in deterioration in quality of life and, in some cases, days lost from school or work. That asthma is common and increasingly prevalent magnifies the importance of any potential economic costs, and promoting asthma control represents an important public health agenda. While lifestyle changes represent a valuable contribution in some patients, the majority of asthmatic patients require therapeutic intervention. The recognition of the role of inflammation in the pathogenesis of asthma has led to an emphasis on regular anti-inflammatory therapy, of which inhaled corticosteroid treatment remains the most superior. In selected patients, further improvements in asthma control may be gained by the addition of regular inhaled long-acting beta(2)-adrenoceptor agonists or oral leukotriene receptor antagonists to inhaled corticosteroid therapy. However, a significant minority of patients with asthma remain poorly controlled despite appropriate treatment, suggesting that additional corticosteroid nonresponsive inflammatory pathways may be operative. Furthermore, some patients with asthma display an accelerated decline in lung function, suggesting that active airway re-modeling is occurring. Such observations have focused attention on the potential to develop new therapies which complement existing treatments by targeting additional inflammatory pathways. The central role of phosphodiesterase (PDE), and in particular the PDE4 enzyme, in the regulation of key inflammatory cells believed to be important in asthma - including eosinophils, lymphocytes, neutrophils and airway smooth muscle - suggests that drugs designed to target this enzyme will have the potential to deliver both bronchodilation and modulate the asthmatic inflammatory response. In vivo studies on individual inflammatory cells suggest that the effects are likely to be favorable in asthma, and animal study models have provided proof of concept; however, first-generation PDE inhibitors have been poorly tolerated due to adverse effects. The development of second-generation agents such as cilomilast and roflumilast heralds a further opportunity to test the potential of these agents, although to date only a limited amount of data from human studies has been published, making it difficult to draw firm conclusions. PMID- 16696591 TI - Flunisolide attenuates nitric oxide-induced DNA damage in rat trachea epithelial cells. AB - In asthma the bronchial epithelium is highly abnormal, with various structural changes. As a consequence, the epithelium becomes an important source of inflammatory mediators that contribute to the ongoing inflammation and remodeling responses occurring in asthma. Compared with normal individuals, the fraction of exhaled nitric oxide (NO) is elevated in patients with asthma, and these levels have been shown to vary with disease activity. Thus, in asthma, epithelial cells may be exposed to large amounts of NO. Increased NO production is associated with the formation of various nitrosating species capable of promoting DNA damage. In this study we investigated the effect of NO on DNA of rat trachea epithelial cells in the presence or absence of flunisolide. Rat airway epithelial cells were prepared and incubated with the NO donor S-nitroso-L-glutathione monoethyl ester (GSNO-MEE). DNA damage was evaluated using single cell gel electrophoresis 'comet assay.' The parameters used as an index of DNA damage were tail length, tail intensity, and tail moment. Results of our study demonstrated that NO induced significant DNA damage in rat airway epithelial cells. Flunisolide in amounts of 11-110 mumol/L significantly reduced all the considered parameters indicating DNA damage. These data indicate that flunisolide may protect epithelial cells from the NO-mediated DNA damage. NO overproduction could contribute to epithelial injury in asthma, and flunisolide seems to attenuate this damage. PMID- 16696593 TI - Real-time in situ monitoring of human prostate photodynamic therapy with diffuse light. AB - Photodynamic therapy (PDT) requires oxygen to cause cellular and vascular tumor damage. Tissue oxygen concentration, in turn, is influenced by blood flow and blood oxygenation. Real-time clinical measurement of these hemodynamic quantities, however, is rare. This paper reports the development and application of a probe, combining diffuse reflectance spectroscopy (DRS) for measurement of tumor blood oxygenation and diffuse correlation spectroscopy (DCS) for measurement of tumor blood flow. The instrument was adapted for clinical use during interstitial prostate PDT. Three patients with locally recurrent prostate cancer received 2 mg/ kg motexafin lutetium (MLu) 3 h before illumination and a total light dose of 100 J/cm(2) at 150 mW/cm. Prostrate blood oxygen saturation (StO2) decreased only slightly (approximately 3%) after treatment. On the other hand, prostate blood flow and total hemoglobin concentration over the course of PDT decreased by 50% and 15%, respectively, suggesting MLu-mediated PDT has an anti-vascular effect. While it is certainly impossible to draw definite conclusions from measurements of only three patients, the observed differences in tumor blood flow and blood oxygenation responses during PDT can, in principle, be used to choose among tissue oxygen consumption models and therefore emphasize the potential clinical value for simultaneous monitoring of both parameters. PMID- 16696594 TI - Insight into the binding of divalent cations to Sepia eumelanin from IR absorption spectroscopy. AB - IR absorption spectroscopy is used to examine the binding of the divalent cations Mg(II), Ca(II), Zn(II) and Cu(II) to melanin granules isolated from the ink sacs of Sepia officinalis. The functional groups of the melanin granules interacting with the bound metal ions are deduced by examining the effect of metal concentration on transition frequencies associated with the COOH, NH and OH moieties of the pigment. The coordinating groups vary with metal ion and with concentration. For the experimental conditions used (initial solution pH of 4, ionic strength of 100 mM and a melanin concentration of 1 mg mL(-1)) Mg(II), Ca(II) and Zn(II) bind to carboxylate groups and Cu(II) binds predominantly to phenolic (catechol) groups However, at a concentration of 10 mM Cu(II) also shows evidence of binding to carboxylate and amine groups, reflecting a secondary binding site that becomes populated as the catechol sites are depleted. PMID- 16696595 TI - Age-dependent photoionization thresholds of melanosomes and lipofuscin isolated from human retinal pigment epithelium cells. AB - Melanosomes and lipofuscin were isolated from 14-, 59-, and 76-year-old, human retinal pigment epithelium specimens and examined. The morphological features of these samples were studied by scanning electron microscopy and atomic force microscopy, and the photoionization properties were examined by photoelectron emission microscopy. Ovoid- and rod-shaped melanosomes were observed. The size of the granules and the distribution between the two shapes show no significant age dependent change. However, there is a higher occurrence of irregularly shaped aggregates of small round granules in older samples which suggests degradation or damage to melanosomes occurs with age. The melanosomes from the 14-year-old donor eye are well characterized by a single photoionization threshold, 4.1 eV, while the two older melanosomes exhibit two thresholds around 4.4 and 3.6 eV. Lipofuscin from both young and old cells show two thresholds, 4.4 and 3.4 eV. The similarity of the potentials observed for aged melanosomes and lipofuscin suggest that the lower threshold in the melanosome sample reflects lipofuscin deposited the surface of the melanosome. The amount, however, is not sufficient to alter the density of the melanosome, and therefore these granules do not separate in a sucrose gradient at densities characteristic of the typical melanolipofuscin granule. These data suggest that thin deposits of lipofuscin on the surface of retinal pigment epithelium melanosomes are common in the aged eye and that this renders the melanosomes more pro-oxidant. PMID- 16696596 TI - No longer 'a private matter': collective action against the manifestations of violence. PMID- 16696597 TI - Peace, love and equality: nurses, interpersonal violence and social justice. PMID- 16696598 TI - Screening for domestic violence: the 'evidence' dilemma. PMID- 16696599 TI - Women defining themselves: shared stories of private hell. AB - In this phenomenological study women who had experienced an abnormal Pap result in the past were asked to share something about themselves as women, prior to describing their experience of having an abnormal Pap result. From their responses, we hear stories of how their sense of self was intricately entwined with their experiences of physical, emotional or sexual violence and/or a profound sense of grief and loss. The stories of violence, trauma and loss alert the nurse and midwife to the possibility that giving a woman information, for example that her Pap test result is abnormal, may compound existing feelings of fear and create a new threat to the woman's sense of 'self'. PMID- 16696600 TI - Lovestruck: women, romantic love and intimate partner violence. AB - Intimate Partner Violence remains a significant problem globally despite health promotion aimed at raising awareness. In particular, there is a current trend for many young women to view some abusive/violent behaviours as acceptable in their relationships. Intimate Partner Violence has serious implications for its short and long term impacts on the health of women and children. Health workers may find working with women a challenging and sometimes frustrating experience. A way forward is to develop clearer understandings of the complexities of Intimate Partner Violence and to better understand women's investments in romantic relationships. In this paper a secondary analysis of data from a narrative study of women's recovery from IPV relationships is presented in order to illustrate discourses that inform underpinnings of romantic relationships. Transcriptions of audio-taped interviews were analysed using a feminist post-structural approach in order to make visible the ways in which the women negotiated their identities in the discourses of femininity. A critical review of current literature was also undertaken to develop the construct of romantic love. Women revealed that cues for Intimate Partner Violence were present early in the relationship but were not recognised at the time. Two positions within the discourse of romantic love were identified that underpinned their desires to establish and invest in the relationship despite the presence of cues for Intimate Partner Violence. These were 'Desperate for a man' and interpreting jealousy as a sign of love. Romantic love may be desirable for the sharing of warmth, safety and protection, and yet can mask behaviours that are cues for domestic violence. Understanding the complex nature of the ways that women's desires are located in the discourse of romantic love has implications for all nurses working to prevent and reduce the incidence of Intimate Partner Violence. PMID- 16696601 TI - Domestic violence, health & health care: women's accounts of their experiences. AB - The current health service response to women's domestic violence related health issues seeks to categorise their presenting symptoms into physical and psychological ailments This gives a narrow and somewhat simplistic focus to what is a complex phenomenon and may obscure women's experiences and limit the opportunities to provide proactive health care. The purpose of this study was to expand knowledge about the health issues of women who are subjected to domestic violence, by affording primacy to women's accounts of health, in order to problematise current services and establish a more woman-centered underpinning for healthcare. The study involved narrative analysis of nine interviews with women who spoke about their health and health issues while they lived with domestic violence. Findings revealed that health was complex, and that health issues were not adequately addressed by health services, in fact health care responses were implicated in further adversely affecting the health of participants. These findings illuminate patterns that underlie individual realities of health and give a foundation from which current services can be evaluated and further developed in ways that may be more sensitive to women's realities and needs. PMID- 16696602 TI - Stories of resistance and healing in the process of leaving abusive relationships. AB - This article examines the unexpected outcomes of a research project that explored the informal support needs of women leaving abusive relationships. The women not only identified informal supports, they also appreciated the time the research offered them to talk about the violent situations, as well as their healing journeys of leaving relationships to sustain their lives without violence. The women's healing journeys were complex and unstable and the feminist approach used in this research empowered participants to acknowledge their abuse, resist the effects of that abuse, and begin their journeys of recovery. The implications of the findings for best nursing practice and models of service delivery are provided. PMID- 16696603 TI - Keeping nurses safe: an industry perspective. PMID- 16696604 TI - Violence, zero tolerance and the subversion of professional practice. AB - Assaults on health care staff have been a fact of life since the earliest years of organised health services, but it is only in recent years that governments have begun to acknowledge the problem. Assaults not only inflict physical and emotional injury, but undermine morale, create a climate of fear, and subvert the quality of care. They are also costly in terms of lost labour, compensation, and legal and procedural expenses. The response to violence in health care settings has thus far ranged from what might be called the 'Ostrich position', in which it is simply ignored, to training in self-defence, the deployment of security staff in clinical areas, conflict resolution training and, more recently, the policy of 'zero tolerance'. This paper examines the rationale for zero tolerance policies, drawing on their origins and applications in the United States and Britain. It suggests that zero tolerance is an ineffective response to violence in health care settings, and its adoption by authorities in Australia should be rejected. It is further argued, that resource allocation and marginalisation are identifiable and modifiable factors contributing to violence in our health care systems. PMID- 16696605 TI - 'They stand you in a corner; you are not to speak': nurses tell of abusive indoctrination in work teams dominated by bullies. AB - This paper reports some of the findings from the first qualitative stage of a large national study of bullying in the nursing workplace currently being undertaken in Australia. The findings reported here reveal how relationships between bullies were embedded within informal organizational alliances, enabling bullies to control work teams and use emotional abuse and psychological violence as a means of enforcing bully-defined 'rules of work'. Within nursing teams, bullies controlled work roles, tasks, and status in the nursing hierarchy through enforcing their 'rules'. Bullies enforced these rules through a process of ritual indoctrination, destroying the self-confidence and self-image of those targeted, and forcing them to eventually resign their position or acquiesce to survive. The merciless, calculated and deliberate nature of the bullying resulted in profound harm for many of those targeted. The findings of this research have implications for the understanding and management of workplace bullying. PMID- 16696606 TI - Workplace violence: the dark side of organisational life. AB - This paper draws on a diverse range of research literature addressing workplace violence, which constitutes one component of the dark side of organisational life. This selective review of the literature has been drawn from the disciplines of nursing, management, psychology and organisational culture. The paper focuses bullying and mobbing in the workplace, addressing its types, causes, the characteristics of bullies and targets and the generalised impact of bullying and mobbing. It also examines whether there are gender issues pertinent to the health care sector. Consideration will also be given to the impact on the individual, group and organization, given the apparent epidemic proportions workplace violence has reached. Ultimately, the question will arise: how can the workplace violence be abolished, specifically within the health care sector, given that we live in a global environment characterised by international bullying (Crawford 1999)? This is a challenge because workplace violence is perpetuated within organisations, due either to cultures of acceptance, or fear of retribution should it be acknowledged and acted upon (or both). PMID- 16696607 TI - Survival of the fittest, or socially constructed phenomena? Theoretical understandings of aggression and violence towards nurses. AB - Violence is an issue that is attracting increasing attention in the nursing literature. There is general agreement that nurses are exposed to unacceptable levels of violence and aggression in the workplace, and that levels of violence are increasing. Despite this attention and awareness, however, violence does not have a standard definition, and theoretical explanations of violence are seldom considered when discussing it in relation to nursing. This paper discusses current issues associated with defining violence, and presents an overview of some of the traditional and contemporary theories of aggression and violence in relation to nursing and the health context. Conflicting issues surrounding predictors and precipitators of violence are examined. We explore the notion that violence is 'part of the job' in nursing. In conclusion, we assert that the current discourses on aggression and violence maintain the status quo, and argue that new explanatory positions on aggression and violence towards nurses are needed. PMID- 16696608 TI - Aggression in nursing: a time to look closely at aged care? PMID- 16696609 TI - Alert but not alarmed? The rhetoric of terrorism and life after 9/11. AB - Our contemporary times are marked by an ever-present regime of sporadic and seemingly escalating violence and counter-violence as terrorism never rests in its campaign to destroy life and property in virtually every corner of the globe. Since September 11, 2001 our political and cultural climate has become increasingly febrile as governments and their various agencies of control ramp up their rhetoric on terrorism with often devastating social and inter-subjective consequences. In this paper I explore the idea of terrorism as a strategic device deployed by a range of actors and entities to manipulate and undermine the Western 'way of life'. Drawing on a diverse range of scholarship I interrogate the politics of fear and anxiety terrorism is designed to propagate and in closing, suggest some tactics ordinary citizens might adopt in order not to be cowed by terrorism's relentless assault on our intellects and sensibilities. PMID- 16696610 TI - Interpersonal violence and mental illness: a literature review. AB - There is a perception that people with a mental illness are dangerous. However, there are still arguments in the research literature as to whether the evidence supports this perception. The major aim of this paper is to review the findings of these studies in regard to the risk of violent behaviour in people with mental illness. An additional aim is to give an overview of the risk factors for violence in people with a mental illness. This systematic search of the literature resulted in good evidence that diagnoses such as schizophrenia and personality disorder are associated with an increased risk of violent behaviour. Substance abuse was the risk factor most associated with an increase in the risk of violent behaviour in people with a mental illness. However, there are substantial differences in the methods used in studies of the risk in violence in people with mental illness resulting in a large variability in the estimates of risk. One of the major causes of variation may be due to the different definitions of violence that are used. The need remains, therefore, for a meta analysis of this literature based on clear definitions of violence in order to get a more accurate estimate of the risk of violence in people with a mental illness. PMID- 16696611 TI - Working with families experiencing violence: the Ploeger Model of enhanced maternal and child health nursing practice. AB - This paper describes the Ploeger Model of enhanced maternal and child health (EMCHN) nursing practice that has been operationalised since 1997. These EMCHNs work with families whose infants and children are exposed to violence. Family violence is complex and many layered. It presents a situation of fear and unpredictability that is difficult to identify and manage. If this situation continues during the child rearing years the effect on the health and wellbeing of infants and children may range from death to life long disabilities. Maternal, siblings and families' health and wellbeing can also be impacted upon for life. The need for enhanced skills for identifying and supporting vulnerable families and safeguarding children at risk has been highlighted by the Department of Health (2004). The Ploeger Model provides an example of early identification and intervention using both secondary and tertiary prevention strategies to identify and make referrals within a multidisciplinary, collaborative partnership, and a process of mutual clinical supervision. Another feature of the model is 'assertive outreach', which assists with early engagement with the family focusing on social support. Each of these helps achieve positive outcomes for children and their families. Such front line professionals must have the knowledge, skills, time and management support to work effectively with these families. PMID- 16696612 TI - Child-to-mother violence: a pilot study. AB - There is vigorous research and debate into family violence. However, there is a paucity of enquiry into violence directed from child-to-mother. This paper presents preliminary results from the pilot of a larger study that examines child to-mother violence in the western suburbs of Sydney Australia. The data are from questionnaires distributed to women who are or have been mothers of children aged 10 to 24 years. Descriptive analysis revealed that an overwhelming majority of the 129 women surveyed experienced some form of threatening behaviour or acts of violence from their child/ren. The main perpetrators were sons and targets mothers. More than half the women had not spoken to anyone about the behaviour previously and a number of women stated their child had witnessed violence in the home. Findings suggest the complexity for women is increased as women are often encouraged to separate from violent partners but their role as mothers prevents them from dividing from a violent child. The incidence appears high, however further evidence from larger samples is required to reveal the full extent of this important social issue. PMID- 16696613 TI - Nurses' understanding of domestic violence. AB - Domestic violence is considered to be an important health care issue world wide. In order to provide appropriate support and treatment to women who have been abused, it is essential that nurses understand and recognise domestic violence. This study aimed to explore nurses' understanding of domestic violence issues utilising a grounded theory approach. A total of 41 emergency nurses was interviewed in selected Australian and Japanese emergency departments. Analysis of data identified six groups of nurse perceivers of domestic violence: nave perceivers, inexperienced perceivers, compassionate perceivers, proactive perceivers, acting perceivers and reluctant perceivers. Findings indicate that the provision of ongoing education and instigation of policy and protocol enable nurses to respond in a structured manner when they encounter women who have been abused. As a result, nurses can contribute to the amelioration of violence in society and enhance the wellbeing of these women. PMID- 16696614 TI - Recognising, responding and resisting violence: a critical challenge for nurses. PMID- 16696615 TI - The power of local alcohol prevention and the Trelleborg Project in southern Sweden. PMID- 16696616 TI - The first Pan American Conference on Alcohol Public Policies and its significance for the region. PMID- 16696617 TI - Moving beyond the transtheoretical model. PMID- 16696619 TI - Genetic epidemiology of cannabis use, abuse and dependence: a comment on Agrawal and Lynskey (2006). PMID- 16696620 TI - The genetics of cannabis involvement in humans: a genetic epidemiological perspective. PMID- 16696622 TI - Thailand: alcohol today. PMID- 16696623 TI - Images from the Brown University Alcohol and Addiction Studies collections. PMID- 16696624 TI - The genetic epidemiology of cannabis use, abuse and dependence. AB - AIMS: The genetic etiology of cannabis use, abuse and dependence has elicited significant interest from genetic epidemiologists. METHODS: Genetically informative studies, including family, twin and adoption studies that have examined the role of genetic and environmental influences on the various stages of cannabis involvement, and the genetic relationship between cannabis, licit drugs and other hard drugs, are reviewed. RESULTS: Findings across a number of such studies have indicated that there is a genetic basis to each stage of cannabis involvement although a proportion of the genetic factors influencing individual stages may be specific to that stage. Multivariate analyses that explore the association between cannabis and licit (alcohol and tobacco) as well as hard drugs (e.g. cocaine, opioids), using multiple methodological strategies, suggest the role of common genetic and environmental influences influencing the liability to cannabis and other drug involvement. CONCLUSIONS: The substantial evidence for the heritability of cannabis use, abuse and dependence underscore the importance of linkage and association studies that aim to find genes of etiologic significance. PMID- 16696625 TI - A community action programme for reducing harmful drinking behaviour among adolescents: the Trelleborg Project. AB - AIMS: To evaluate a 3-year community intervention programme by measuring changes in drinking patterns in a 15-16-year-old population. DESIGN, SETTING, PARTICIPANTS AND MEASUREMENTS: The action programme included five demand-reducing and one supply-reducing interventions. Cross-sectional, non-repeated data were collected from a questionnaire distributed in classrooms from 1999 to 2001 and 2003 (n = 1376, 724 boys and 652 girls; response rate = 92.3%). Stepwise logistic regression analyses were used to determine the relationship between different risk factors and excessive drinking, heavy episodic drinking, purchaser of alcohol and alcohol provided by parents. The results from the intervention community were also compared with similar Swedish cross-sectional data sets. FINDINGS: The results of our analyses indicated a decrease in harmful drinking behaviour in Trelleborg when comparing baseline with postintervention measurements. The comparison with other studies showed that the changes in these indicators were more rapid and consistent in Trelleborg. Finally, the multivariable logistic regression analyses showed that the outcomes were not likely to be attributed to changes in environmental factors. CONCLUSIONS: We concluded that a community action programme based on the systems approach reduced hazardous alcohol consumption effectively among adolescents in Trelleborg. PMID- 16696626 TI - Estimating the effect of help-seeking on achieving recovery from alcohol dependence. AB - AIMS: To investigate the effect of help-seeking on the likelihood of recovery from Diagnostic and Statistical Manual version IV (DSM-IV) alcohol dependence, specifically examining the impact of model selection, factors that moderate the effect of help-seeking and distinctions between the effects of 12-Step participation and formal treatment. DESIGN: This analysis is based on data from the Wave 1 2001-02 National Epidemiologic Survey on Alcohol and Related Conditions (NESARC), a cross-sectional, retrospective survey of a nationally representative sample of US adults 18 years of age and over. The analytical sample consisted of 4422 individuals with prior-to-past-year (PPY) onset of DSM IV alcohol dependence. METHODS: Logistic regression, proportional hazards and time-dependent proportional hazards models were used to estimate the effects of help-seeking on three outcomes: (1) any recovery from alcohol dependence, which required full remission of all symptoms of alcohol abuse and dependence and excluded asymptomatic drinkers whose alcohol consumption exceeded low-risk drinking guidelines; (2) non-abstinent recovery (NR), representing low-risk asymptomatic drinkers; and (3) abstinent recovery (AR), representing abstainers. FINDINGS: Only one-quarter of individuals with PPY-onset alcohol dependence had ever sought help for alcohol problems, including 3.1% who had participated in 12 Step programs only, 5.4% who had received formal treatment only and 17.0% with both 12-Step and formal treatment. Based on the most appropriate model, help seeking increased the likelihood of any recovery [hazard rate ratio (HRR) = 2.38], NR (HRR = 1.50) and AR (HRR = 4.01). The impact of help-seeking on AR did not show any significant variation across the exposure period but was modified by severity among other factors. Individuals who participated in 12-Step programs in addition to formal treatment had almost twice the chance of recovery and more than more than twice the chance of AR compared with those who received formal treatment alone. CONCLUSIONS: Help-seeking plays a significant role in the achievement of abstinent recovery from alcohol dependence, with 12-Step participation playing a major role. Appropriate model selection is critical to assessing the impact of help-seeking. PMID- 16696627 TI - Trends in alcohol-related mortality in Sweden 1969-2002: an age-period-cohort analysis. AB - AIM: To study the effects of age, period and cohorts on alcohol-related mortality trends in Sweden. DESIGN: The study comprises an age-period-cohort analysis. SETTING AND PARTICIPANTS: The analysis was based on all deaths in the Swedish population between 1969 and 2002. MEASUREMENTS: Data on alcohol-related deaths in Sweden from 1969 to 2002 excluding accidental injury and homicide were used. The analysis covered 43 021 deaths. FINDINGS: Time period and birth cohort both influenced alcohol-related mortality. Male cohorts born in the 1930-40s exhibited the highest alcohol-related mortality, while for females those born in the 1940 50s had the highest alcohol-related mortality. For both men and women, those born in the 1960-70s had the lowest age-adjusted alcohol-related mortality. High-risk cohorts were young or in early adulthood during the periods that alcohol became more available in Sweden. The low-risk cohorts of the 1960-70s were brought up during a period when society was concerned with increasing alcohol problems and more emphasis was placed on issuing alcohol awareness information in schools. CONCLUSIONS: Cohort effects were found suggesting that the link between alcohol consumption and non-accident alcohol-related mortality at the population level is dependent on other factors that may change over time. One such factor may be that restrictive alcohol policies have a greater effect on drinking in those who are younger at the time they are put into effect. PMID- 16696628 TI - A longitudinal evaluation of medical student knowledge, skills and attitudes to alcohol and drugs. AB - AIM: To examine the knowledge, skills and attitudes of medical students to alcohol and drugs as training progresses. DESIGN: A longitudinal, prospective, cohort-based design. SETTING: The four schools of medicine in New Zealand. PARTICIPANTS: All second-year medical students (first year of pre-clinical medical health sciences) in New Zealand were administered a questionnaire which was repeated in the fourth (first year of significant clinical exposure) and then sixth years (final year). A response rate of 98% in the second year, 75% in the fourth year and 34% in the sixth year, with a total of 637 respondents (47.8% male) and an overall response rate of 68%. QUESTIONNAIRE: The questionnaire consisted of 43 questions assessing knowledge and skills -- a mixture of true/false and scenario stem-based multiple-choice questions and 25 attitudinal questions scored on a Likert scale. Demographic questions included first language, ethnicity and personal consumption of alcohol and tobacco. FINDINGS: The competence (knowledge plus skills) correct scores increased from 23.4% at the second year to 53.6% at the fourth year to 71.8% at the sixth year, being better in those students who drank alcohol and whose first language was English (P < 0.002). As training progressed the student's perceptions of their role adequacy regarding the effectiveness of the management of illicit drug users diminished. For example, at second year 21% and at sixth year 51% of students felt least effective in helping patients to reduce illicit drug use. At the sixth year, 15% of sixth year students regarded the self-prescription of psychoactive drugs as responsible practice. CONCLUSION: Education on alcohol and drugs for students remains a crucial but under provided part of the undergraduate medical curriculum. This research demonstrated that while positive teaching outcomes were apparent, further changes to medical student curricula need to be considered to address specific knowledge deficits and to increase the therapeutic commitment and professional safety of medical students to alcohol and drugs. PMID- 16696629 TI - Violent victimization and drug involvement among Mexican middle school students. AB - AIMS: To answer the following research questions: (a) is there an association between violent victimization and exposure to opportunities to use marijuana, inhalants and cocaine and (b) is there an association between violent victimization and actual drug use among youth with drug-using opportunities? DESIGN: Cross-sectional survey. SETTING: Two middle schools located in the Historic Downtown area of Mexico City. PARTICIPANTS: The entire body of students (n = 767; mean age 13.8 years, 52% males). MEASUREMENTS: Qualitative research was used to develop questions on drug exposure opportunities and violent victimization. Standardized questions on life-time alcohol, tobacco, marijuana, inhalant drugs and cocaine use were also included, as well as questions on violent victimization and other covariates. FINDINGS: One-quarter (25%) of students had an opportunity to try marijuana, inhalant drugs or cocaine; 35% who had an opportunity actually used at least one drug. In this sample, 59% had been victimized violently. Youth who had been victimized had greater odds of opportunities to use drugs compared to those who had not been victimized [adjusted odds ratio (OR) = 3.8; 95% confidence interval (CI), 2.4, 6.1]. Once exposure opportunity is taken into consideration, no association was evident between violent victimization and actual drug use (adjusted OR = 0.9; 95% CI, 0.4, 2.1). CONCLUSIONS: It is possible to trace back the association between violent victimization and drug use to differences in exposure to opportunities. Limitations considered, this study suggests interventions to improve micro and macro contexts, such as families, schools and communities, so young people can have better places to live and develop. PMID- 16696630 TI - Public sector managed care: a comparative evaluation of substance abuse treatment in three counties. AB - AIMS: A study of publicly funded substance abuse treatment systems compared MidState, a county that reorganized its treatment system using managed care principles, to two other California counties that took different approaches, NorthState and SouthState. It was hypothesized that MidState would have better outcomes due to its emphasis on quality of care. DESIGN: This natural experiment compared the 'experimental' county, MidState, to two 'control' counties, assessing client outcomes following treatment. Administrative and historical exigencies that may affect system differences were explored in interviews with treatment program managers and staff. SETTINGS: Comparison counties were selected using treatment system and county census data, maximizing similarities to enhance internal validity. PARTICIPANTS: Adult clients (n = 681) were interviewed when beginning treatment and 12 months later (81% response rate). In addition, 50 treatment program managers and staff members across the three counties were interviewed during the year of client recruitment. MEASUREMENTS: Client interviews assessed functioning in the seven Addiction Severity Index domains alcohol, drug, psychiatric, legal, employment, medical and family/social. FINDINGS: Outcomes (differences between baseline and 12 month composite scores) did not differ between counties in six of seven domains; in the seventh, psychiatric functioning, SouthState had better outcomes than MidState. Staff interviews indicated generally similar treatment strategies across counties, with MidState supplying greater oversight and performance standards. CONCLUSIONS: Managed care in public sector treatment generally did not result in poorer outcomes. Future attention in MidState to the barriers to successful implementation of individualized treatment, and to dual diagnosis treatment, might bring more positive results. PMID- 16696631 TI - The use of record linkage to examine illicit drug use in pregnancy. AB - AIMS: To examine the obstetric and perinatal outcomes for women with a drug related hospital admission during pregnancy. DESIGN: Antenatal and birth admissions to New South Wales (NSW) hospitals from the NSW Inpatient Statistics Collection were linked to birth information from the NSW Midwives Data Collection over a 5-year period (1998-2002). MEASUREMENTS: Birth admissions were flagged as positive for drug use where a birth admission or any pregnancy admission for that birth involved an opioid, cannabis or stimulant-related International Classification of Diseases version 10 Australian modification (ICD-10AM) code. FINDINGS: A total of 416 834 live births were analysed over a 5-year period (1998 2002). Of these, 1974 pregnancies had an opioid ICD-10AM diagnosis recorded, 552 a stimulant-related ICD-10AM and 2172 a cannabis ICD-10AM diagnosis. Births in each of the drug groups were to women who were younger, had a higher number of previous pregnancies, were indigenous, smoked heavily and were not privately insured. These women also presented later in their pregnancy to antenatal services and were more likely to arrive at hospital unbooked. Neonates born to women in each of the drug groups were more likely to be premature and were admitted to neonatal intensive care and special care nursery more often, with neonates born to women in the opioid group admitted most often. CONCLUSIONS: Linked population level administrative data is a powerful method for examining the maternal and neonatal outcomes associated with the use of specific illicit drugs during pregnancy. PMID- 16696632 TI - The effects of nicotine gum and counseling among African American light smokers: a 2 x 2 factorial design. AB - AIM: Approximately 50% of African American smokers are light smokers (smoke < or = 10 cigarettes a day). The prevalence of light smoking in the United States is increasing, yet there has not been a single smoking cessation clinical trial targeting light smokers. The purpose of this 2 x 2 factorial, randomized clinical trial was to evaluate the efficacy of nicotine gum (2 mg versus placebo) and counseling (motivational interviewing versus health education) for African American light smokers. DESIGN: Participants were assigned randomly to one of four study arms: 2 mg nicotine gum plus health education (HE); 2 mg nicotine gum plus motivational interviewing (MI); placebo gum plus HE; and placebo gum plus MI. PARTICIPANTS AND SETTING: A total of 755 African American light smokers (66% female, mean age = 45) were enrolled at a community health center over a 16-month period. INTERVENTION AND MEASUREMENTS: Participants received an 8-week supply of nicotine gum and six counseling sessions during the course of the 26-week study. Biochemical measures included expired carbon monoxide (CO) and serum and salivary cotinine. FINDINGS: Seven-day quit rates for nicotine gum were no better than for the placebo group (14.2% versus 11.1%, P = 0.232) at 6 months. However, a counseling effect emerged, with HE performing significantly better than MI (16.7% versus 8.5%, P < 0.001). These results were consistent across outcome time-points (weeks 1, 8, and 26). CONCLUSIONS: Results highlight the potential positive impact of directive information and advice-oriented counseling on smoking cessation. Studies are needed to assess other interventions that may further improve quit rates among African American light smokers who are motivated to quit. PMID- 16696634 TI - Russia, substance misuse and social transition. PMID- 16696633 TI - Nicotine dependence is associated with compulsive alcohol craving. AB - AIMS: To investigate a possible association of nicotine dependence and alcohol craving. DESIGN: A prospective cross-sectional study on patients diagnosed with alcohol dependence. SETTING: Detoxification unit of a regional psychiatric hospital. PARTICIPANTS: A total of 127 smoking male patients were included in the study at admission for detoxification from alcohol. MEASUREMENTS: The Fagerstrom Test for Nicotine Dependence (FTND) was used to assess the severity of nicotine dependence while the Obsessive Compulsive Craving Scale (OCDS) was used to measure alcohol craving. The OCDS was assessed at admission and after 7 days of withdrawal treatment, distinguishing the total score, the obsessive and the compulsive subscale. FINDINGS: Spearman's correlation revealed a significant association between the extent of alcohol craving and the FTND score (day 0, n = 127: OCDS total score r = 0.238, P = 0.007; OCDS compulsive score r = 0.280, P = 0.001; day 7; n = 94: OCDS total score r = 0.212, P = 0.040; OCDS compulsive score r = 0.225, P = 0.029). CONCLUSIONS: The severity of nicotine dependence is associated with higher craving in alcohol-dependent patients. These results point towards shared pathophysiological mechanisms in alcohol craving and nicotine addiction. PMID- 16696635 TI - Dependence on zopiclone. PMID- 16696641 TI - Cryptococcus: the once-sleeping giant is fully awake. PMID- 16696642 TI - Cryptococcus neoformans: the yeast that likes it hot. AB - The ability of fungi to grow well at mammalian body temperatures is a fundamental characteristic of invasive human fungal pathogens. Cryptococcus neoformans, with its genetics, molecular biology, robust animal models and clinical importance, has become a premier fungal model system for molecular fungal pathogenesis studies. There has been over a half century of study into how C. neoformans grows at high temperatures. However, recently the understanding of high-temperature growth at a molecular level has dramatically accelerated. Many strategies have been used to identify genes and over a dozen genes have already been identified to be necessary for high-temperature growth. It is likely that there are many more to discover. It is clear that, as further studies understand how this encapsulated yeast is able to withstand the stresses of high temperature at a genetic and molecular basis, we will also know more about how it and other fungal pathogens have evolved into well-established human pathogens. PMID- 16696643 TI - Lipid metabolism in Cryptococcus neoformans. AB - In recent years, lipids have been shown to act as signalling molecules not only in mammalian cells but also in many other eukaryotes. Whereas in mammalian cells lipids regulate cellular functions that play crucial roles in the regulation of pathobiological processes, such as cancer, cardiovascular and neurodegenerative disorders, and inflammation, in the fungus Cryptococcus neoformans lipids play key roles in the regulation of pathogenic traits required for the development of cryptococcosis, an infectious disease particularly frequent in immunocompromised individuals. In this minireview we discuss recent advances in the understanding of lipid metabolism in this important human pathogen, highlighting the potential of fungal lipid enzymatic pathways as promising new drug targets. PMID- 16696644 TI - Phenotypic switching and its implications for the pathogenesis of Cryptococcus neoformans. AB - Phenotypic switching has been described in several strains of Cryptococcus neoformans. It occurs in vivo during chronic infection and is associated with differential gene expression and changes in virulence. The switch involves changes in the polysaccharide capsule and cell wall that affect the yeast's ability to resist phagocytosis. In addition, the phenotypic switch variants elicit qualitatively different inflammatory responses in the host. The host's immune response ultimately affects selection of the switch variants in animal models of chronic cryptococcosis. The biological relevance of phenotypic switching is demonstrated in several murine infection models and further underlines the importance of phenotypic switching in the setting of human disease. This includes the association of switching and poor outcome in chronic infection, the ability of the mucoid variant of strain RC-2 (RC-2 MC) but not the smooth variant (RC-2 SM) to promote increased intracranial pressure in a rat model, and lastly the observation that antifungal interventions can promote the selection of more virulent switch variants during chronic murine infection. PMID- 16696645 TI - Intersection of fungal fitness and virulence in Cryptococcus neoformans. AB - The use of insertional mutagenesis to discover genes that impact laccase activity has resulted in the identification of multiple cellular processes that affect the fitness of Cryptococcus neoformans. Fitness has been defined as the ability of an organism to propagate and evolve within a given environment. Because the human host is an evolutionary dead-end for an opportunistic pathogen, we have defined pathogenic fitness here as the capability to successfully propagate within the stressful environment of the host, causing disease by expression of virulence traits that damage the host. In this review, laccase-deficient insertional mutants will be highlighted in terms of the basic biological processes in which they are involved. The impact of laccase-associated cellular functions on fitness and virulence will be discussed, as will the mutants' potential as therapeutic targets. Vacuolar function, copper homeostasis, mitochondrial function and carbon repression are covered. PMID- 16696646 TI - Glycosyltransferases and their products: cryptococcal variations on fungal themes. AB - Glycosyltransferases are specific enzymes that catalyse the transfer of monosaccharide moieties to biological substrates, including proteins, lipids and carbohydrates. These enzymes are present from prokaryotes to humans, and their glycoconjugate products are often vital for survival of the organism. Many glycosyltransferases found in fungal pathogens such as Cryptococcus neoformans do not exist in mammalian systems, making them attractive potential targets for selectively toxic agents. In this article, we present the features of this diverse class of enzymes, and review the fungal glycosyltransferases that are involved in synthesis of the cell wall, the cryptococcal capsule, glycoproteins and glycolipids. We specifically focus on enzymes that have been identified or studied in C. neoformans, and we consider future directions for research on glycosyltransferases in the context of this opportunistic pathogen. PMID- 16696647 TI - The molecular basis for the immunogenicity of Cryptococcus neoformans mannoproteins. AB - T-cell-mediated immunity is necessary for effective host defenses against infections caused by Cryptococcus neoformans. Clinical and experimental studies have identified a heterogeneous family of mannoproteins as critical cryptococcal antigens responsible for stimulating T-cell responses. The archetypal mannoprotein has a signal sequence, a functional domain, a serine/threonine-rich region and a site for attachment of a glycosylphosphatidylinositol anchor. Extensive O-mannosylation, which occurs at the serine/threonine region, facilitates recognition by mannose receptors on antigen-presenting cells, particularly dendritic cells. This results in efficient antigen uptake, processing and presentation to T cells. Inhibition of mannose receptors or deglycosylation of mannoproteins profoundly inhibits T-cell responses, demonstrating the crucial contribution of mannosylation to immunogenicity. PMID- 16696648 TI - Towards a vaccine for Cryptococcus neoformans: principles and caveats. AB - In the Damage-response framework of microbial pathogenesis, infectious diseases are one outcome of a host-microorganism interaction in a susceptible host. In cryptococcal disease, damage to the host is caused by Cryptococcus neoformans virulence determinants, the nature of the host response, or both. Further, the disease may be acute or reactivated from a latent state. Hence, a vaccine for C. neoformans would need to prevent disease resulting from either acute or reactivated infection. The evidence to support the development of a vaccine for C. neoformans that induces antibody-mediated immunity is discussed herein. PMID- 16696649 TI - Glucuronoxylomannan exhibits potent immunosuppressive properties. AB - Cryptococcus neoformans is an opportunistic fungus responsible for life threatening infections in immunocompromised and occasionally in immunocompetent hosts. The fungus is endowed with several virulence factors such as its capsular polysaccharide, which plays a key role in virulence. The major component of capsular material of C. neoformans is glucuronoxylomannan, a polysaccharide that exhibits potent immunosuppressive properties in vitro and in vivo. Here we describe a new aspect relative to glucuronoxylomannan-induced immunosuppression. In this review we analyze the suppressive effects of glucuronoxylomannan and the mechanisms leading to induction of apoptosis in T cells. PMID- 16696650 TI - Contemplating the murine test tube: lessons from natural killer cells and Cryptococcus neoformans. AB - Murine experimentation has provided many useful tools, including the ability to knockout or over-express genes and to perform experiments that are limited by ethical considerations. Over the past century, mice have imparted valuable insights into the biology of many systems, including human immunity. However, although there are many similarities between the immune response of humans and mice, there are also many differences; none is more prominent than when examining natural killer cell biology. These differences include tissue distribution, effector molecules, receptor repertoire, and cytokine responses, all of which have important implications when extrapolating the studies to the human immune responses to Cryptococcus neoformans. PMID- 16696651 TI - Application of proton nuclear magnetic resonance spectroscopy to the study of Cryptococcus and cryptococcosis. AB - Proton nuclear magnetic resonance spectroscopy is a nondestructive technique that identifies chemicals in solution and in living cells. It has been used in cryptococcal research to identify the primary structure of capsular glucuronoxylomannans, link cellular apoptosis susceptibility (CAS) genes to positioning of residues on the mannose backbone of glucuronoxylomannan, and verify that the cryptococcal virulence determinant, phospholipase B, is elaborated in vivo. Promising clinical applications include speciation (Cryptococcus neoformans and Cryptococcus gattii), with preliminary evidence that varieties neoformans and grubii can also be distinguished, non-invasive diagnosis of cerebral cryptococcomas, and, in cases of meningitis, monitoring therapeutic response by analysis of cerebrospinal fluid. PMID- 16696652 TI - The pursuit of cryptococcal pathogenesis: heterologous hosts and the study of cryptococcal host-pathogen interactions. AB - Analysis of the molecular mechanisms by which a pathogen interacts with the human host is most commonly performed using a mammalian model of infection. However, several virulence-related genes previously shown to be involved in mammalian infection with Cryptococcus neoformans have also been shown to play a role in the interaction of these pathogens with invertebrates, such as Acanthamoeba castellanii, Caenorhabditis elegans, Dictyostelium discoideum, Drosophila melanogaster and Galleria mellonella. The study of host-pathogen interactions using these model hosts has allowed rapid screening of mutant libraries and can be used for the study of evolutionarily preserved aspects of microbial virulence and host response. PMID- 16696653 TI - Do major species concepts support one, two or more species within Cryptococcus neoformans? AB - Cryptococcus neoformans, the agent of cryptococcosis, had been considered a homogeneous species until 1949 when the existence of four serotypes was revealed based on the antigenic properties of its polysaccharide capsule. Such heterogeneity of the species, however, remained obscure until the two morphologically distinct teleomorphs of C. neoformans were discovered during the mid 1970s. The teleomorph Filobasidiella neoformans was found to be produced by strains of serotype A and D, and Filobasidiella bacillispora was found to be produced by strains of serotype B and C. Ensuing studies revealed numerous differences between the anamorphs of the two Filobasidiella species with regard to their ecology, epidemiology, pathobiology, biochemistry and genetics. At present, the etiologic agent of cryptococcosis is classified into two species, C. neoformans (serotypes A and D) and Cryptococcus gattii (serotypes B and C). Intraspecific genetic diversity has also been revealed as more genotyping methods have been applied for each serotype. As a result, the number of scientifically valid species within C. neoformans has become a controversial issue because of the differing opinions among taxonomists as to the appropriate definition of a species. There are three major species concepts that govern classification of organisms: phenetic (morphologic, phenotypic), biologic (interbreeding) and cladistic (evolutionary, phylogenetic). Classification of the two C. neoformans species has been based on the phenetic as well as the biologic species concept, which is also supported by the cladistic species concept. In this paper, we review and attest to the validity of the current two-species system in light of the three major species concepts. PMID- 16696654 TI - Looking for sex in the fungal pathogens Cryptococcus neoformans and Cryptococcus gattii. AB - Why are we interested in understanding the mode of reproduction being used by the fungal pathogens Cryptococcus neoformans and Cryptococcus gattii? Empirical evidence has finally supported the long-held assumption that, by increasing the rate of adaptive evolution, sex increases the chances of long-term survival. Understanding the ability of pathogenic organisms to adapt to diagnostic and treatment regimes is also important in the fight against the diseases caused by these organisms. This review looks at the different approaches used to identify population structure in C. neoformans and C. gattii. These are sexual species; however, recombination in natural populations has only recently been found. We highlight the importance of population selection and the value of both indirect molecular analysis and direct biological evidence for sexual recombination, when looking for the mode of reproduction in these fungal pathogens. PMID- 16696655 TI - Unique hybrids between the fungal pathogens Cryptococcus neoformans and Cryptococcus gattii. AB - Cryptococcus neoformans and Cryptococcus gattii are yeasts that cause meningoencephalitis, but that differ in host range and geographical distribution. Cryptococcus neoformans occurs world-wide and mostly infects immunocompromised patients, whereas C. gattii occurs mainly in (sub)tropical regions and infects healthy individuals. Anomalous C. neoformans strains were isolated from patients. These strains were found to be monokaryotic, and diploid or aneuploid. Amplified Fragment Length Polymorphism (AFLP) and sequence analyses indicated that AFLP genotypes 2 (C. neoformans) and 4 (C. gattii) were present. The strains were serologically BD. Mating- and serotype-specific PCR reactions showed that the strains were MATa-serotype D/MATalpha-serotype B. This study is the first to describe naturally occurring hybrids between C. neoformans and C. gattii. PMID- 16696656 TI - Cryptococcus neoformans population includes hybrid strains homozygous at mating type locus. AB - Recent attempts to characterise the hybrid strains of Cryptococcus neoformans have led to the identification of a cryptic population of hybrid strains ('H strains') with double DNA content but only a single mating-type allele. To verify a set of hypotheses concerning their origin, we investigated 14 previously isolated H strains and ten F1-progeny strains arising from H99 and JEC20 mating. The double DNA content was tested by flow cytometry; the presence of only one mating type was tested by amplifying 12 mating-type-specific genes and one gene unlinked with the mating-type locus (URA5). Analysis of the F1 progeny identified two H strains, and electrophoretic karyotyping confirmed the occurrence of genetic recombination. The simultaneous presence of the homozygous and heterozygous loci, and the fact that all of the F1-progeny strains presented a recombinant karyotype, suggest that the H strains originated from the post meiotic random fusion of two of the four recombinant nuclei. Further studies are required to elucidate the role of the homozygous mating-type loci in the virulence of C. neoformans. PMID- 16696657 TI - Molecular analysis of 311 Cryptococcus neoformans isolates from a 30-month ECMM survey of cryptococcosis in Europe. AB - During a European Confederation of Medical Mycology (ECMM) prospective survey of cryptococcosis in Europe (from July 1997 to December 1999) 655 cases were reported from 17 countries; 565 of the completed questionnaires were evaluable. Cryptococcosis was associated with HIV infection in 77% of cases (range 57.5 94%). Assessment of the laboratory data highlighted the lack of defined standard procedures for the diagnosis of cryptococcosis: the antigen test was not usually used for screening, the disease was mainly recognised when meningitis occurred (65% of patients) and, with the exception of a few cases, the extent of the infection was not investigated. Cryptococcus neoformans was the etiological agent in all of the cases except for six caused by C. gattii and four by other Cryptococcus species. A total of 311 C. neoformans strains were serotyped by Crypto Check latex agglutination, genotyped by PCR-fingerprinting using the (GACA)4 oligonucleotide as a single primer, and their mating type was determined by PCR of the STE20 alleles. Serotype A was the most represented (51% of the isolates), followed by serotype D (30%) and serotype AD (19%). PCR-fingerprinting analysis significantly increased the percentage of hybrid strains to 30%, as 6% of the serotype A and 28% of the serotype D isolates were of the VN3 or VN4 hybrid genotype. In addition, the mating type determinations revealed the MATa serotype A allele in one haploid strain and 28 hybrids, and hybrid isolates with a single mating type (four Aalpha and two Dalpha) were also identified. This is the first prospective survey to be carried out in Europe which has attempted to investigate the epidemiology of cryptococcosis and the population structure of C. neoformans, and the results obtained thus far show the widespread involvement of AD hybrid strains in C. neoformans infections. PMID- 16696658 TI - Yeast diversity sampling on the San Juan Islands reveals no evidence for the spread of the Vancouver Island Cryptococcus gattii outbreak to this locale. AB - Biological diversity has been estimated for various phyla of life, such as insects and mammals, but in the microbe world is has been difficult to determine species richness and abundance. Here we describe a study of species diversity of fungi with a yeast-like colony morphology from the San Juan Islands, a group of islands that lies southeast of Vancouver Island, Canada. Our sampling revealed that the San Juan archipelago biosphere contains a diverse range of such fungi predominantly belonging to the Basidiomycota, particularly of the order Tremellales. One member of this group, Cryptococcus gattii, is the etiological agent of a current and ongoing outbreak of cryptococcosis on nearby Vancouver Island. Our sampling did not, however, reveal this species. While the lack of recovery of C. gattii does not preclude its presence on the San Juan Islands, our results suggest that the Strait of Juan de Fuca may be serving as a geographical barrier to restrict the dispersal of this primary human fungal pathogen into the United States. PMID- 16696659 TI - Molecular epidemiology of clinical and environmental isolates of the Cryptococcus neoformans species complex reveals a high genetic diversity and the presence of the molecular type VGII mating type a in Colombia. AB - The aim of this study was to investigate the epidemiological relationships of clinical and environmental isolates of the Cryptococcus neoformans species complex in Colombia. The current study reflects data from 1987 to 2004. In Colombia serotypes A and B are most frequently recovered from patients and the environment. Of the 178 clinical isolates studied, 91.1% were of serotype A, 8.4% serotype B and 0.5% serotype C. Of the 247 environmental isolates, 44.2% were of serotype A, 42.6% serotype B and 13.2% serotype C. No serotype D isolates were isolated. Serotype AD has not been recovered in Colombia. PCR fingerprinting with the primers M13, (GACA)4 and (GTG)5 and URA5 gene restriction fragment length polymorphism analysis grouped the majority of clinical serotype A and environmental serotype B isolates into the molecular types VNI (98.1%) and VGII (100%), respectively. Mating type alpha was determined in 99.3% of serotype A isolates, but 96.6% of serotype B isolates were of mating type a. Similar profiles between clinical and environmental isolates suggest that the patients may have acquired the infection from the environment. The data presented form part of the Colombian contribution to the ongoing global survey of the C. neoformans species complex. PMID- 16696660 TI - Influence of climatic conditions on the isolation of members of the Cryptococcus neoformans species complex from trees in Colombia from 1992-2004. AB - The aim of this retrospective study was to analyze the relationship between occurrence of the serotypes of the Cryptococcus neoformans species complex in tree samples and the climatic conditions registered during samplings in four cities of Colombia, between 1992 and 2004, by means of a logistic regression model and lagged Pearson correlations. During 97 collection dates, 8220 samples from different tree species were taken, of which 2.63% were positive: 56.5% yielded serotype B, 24.7% serotype C and 18.8% serotype A isolates. The prevalence of the serotypes varied among the cities. The results suggest that environmental climatic conditions, mainly humidity, temperature, evaporation and solar radiation, can affect the occurrence of the different serotypes in trees in a differential manner. These different climatic tolerances were reflected in the geographic distribution of the serotypes in Colombia. The climatic conditions for 15 days before the sampling date were correlated with positive or negative isolation of the different serotypes. PMID- 16696661 TI - Identification of major proteins secreted by Cryptococcus neoformans. AB - The characterization of proteins secreted by Cryptococcus neoformans is of relevance to the identification of vaccine candidates, because concentrated supernatants from the fungus have been shown to be immunoprotective in previous studies. After fractionation of supernatants by anion exchange chromatography and preparative electrophoresis, we obtained the N-terminal amino acid sequences of 13 major proteins. Using a C. neoformans nucleotide database, we were able to clone and sequence the ORFs coding for 12 of these proteins. Some of the genes are identical to previously described ones, while six encode novel proteins, including four putative mannoproteins. The molecular characterization of these and other secreted products may provide useful information in the development of immune-based strategies to control cryptococcosis. PMID- 16696662 TI - The Skn7 response regulator of Cryptococcus neoformans is involved in oxidative stress signalling and augments intracellular survival in endothelium. AB - Cryptococcus neoformans is the causative agent of cryptococcal meningoencephalitis. There is accumulating evidence that C. neoformans is a facultative intracellular pathogen, residing in macrophages and endothelium. The molecular mechanism conferring resistance to phagolysosomal killing in these cells is a key unresolved issue. To gain insight into the fungal adaptive strategies, serial analysis of gene expression was used to map genes differentially expressed in an intraphagocytic environment. By comparing transcript profiles of C. neoformans serotype D B3501 cells recovered from endothelial cells with those from free-grown cryptococci, we identified the cryptococcal homologue of the SKN7 two-component stress response regulator gene from Saccharomyces cerevisiae. Studies with C. neoformans cells disrupted for SKN7 revealed an increased susceptibility to t-butyl hydroperoxide (100% lethality at 0.7 mM, vs. 1.0 mM for wild type) and significantly lower survival rates in endothelial infection experiments. Mice experiments revealed that SKN7 disruption strongly attenuates cryptococcal virulence in vivo. We propose that Skn7 (co-)regulates the fungal adaptive strategy, allowing intraphagocytic survival by conferring resistance to phagolysosomal killing in endothelial cells. PMID- 16696663 TI - Equatorial ring-like channels in the Cryptococcus neoformans polysaccharide capsule. AB - Under certain conditions, India ink particles can penetrate the capsule of the opportunistic pathogen Cryptococcus neoformans. India ink penetration gave two distinct patterns, one as a ring in the middle of the capsule, and another as a double spot located at opposite poles of the cells. These spots were perpendicularly orientated to the bud. This pattern suggests the existence of a localized structure deep in the capsule that can accumulate large insoluble particles. Although the mechanisms responsible for the assemblage and maintenance of ring-like channels are not understood, their existence deep within the capsule implies a new level of complexity for this enigmatic structure. PMID- 16696664 TI - Trends and highlights in Candida research. The 8th ASM Conference about Candida and Candidiasis, Denver (Colorado, USA), 13-17 March 2006. PMID- 16696665 TI - Probiotics and their fermented food products are beneficial for health. AB - Probiotics are usually defined as microbial food supplements with beneficial effects on the consumers. Most probiotics fall into the group of organisms' known as lactic acid-producing bacteria and are normally consumed in the form of yogurt, fermented milks or other fermented foods. Some of the beneficial effect of lactic acid bacteria consumption include: (i) improving intestinal tract health; (ii) enhancing the immune system, synthesizing and enhancing the bioavailability of nutrients; (iii) reducing symptoms of lactose intolerance, decreasing the prevalence of allergy in susceptible individuals; and (iv) reducing risk of certain cancers. The mechanisms by which probiotics exert their effects are largely unknown, but may involve modifying gut pH, antagonizing pathogens through production of antimicrobial compounds, competing for pathogen binding and receptor sites as well as for available nutrients and growth factors, stimulating immunomodulatory cells, and producing lactase. Selection criteria, efficacy, food and supplement sources and safety issues around probiotics are reviewed. Recent scientific investigation has supported the important role of probiotics as a part of a healthy diet for human as well as for animals and may be an avenue to provide a safe, cost effective, and 'natural' approach that adds a barrier against microbial infection. This paper presents a review of probiotics in health maintenance and disease prevention. PMID- 16696666 TI - Activity of trans-2-hexenal against Penicillium expansum in 'Conference' pears. AB - AIMS: To investigate the effects of trans-2-hexenal on blue mould disease, patulin content and fruit quality in 'Conference' pears. METHODS AND RESULTS: Fruits, wounded and inoculated with Penicillium expansum or non-inoculated, were exposed to trans-2-hexenal vapour treatment (12.5 microl l(-1)) at 20 degrees C. A greater reduction of decay was obtained by treatment application 24 or 48 h after inoculation, in contrast trans-2-hexenal application 2 h after inoculation was ineffective. Fruit storage temperature (-1 degrees C) after treatment did not affect the antifungal activity. Although 2-h exposure to trans-2-hexenal was effective in reducing blue mould, an exposure of at least 8 h was required to reduce fruit patulin content. Treatments did not affect fruit physical-chemical characteristics. After 6 days at 20 degrees C following exposure, trans-2-hexenal residue in treated fruits was less than the natural content of the compound in unripe fruits. CONCLUSIONS: trans-2-Hexenal treatment is effective in the reduction of blue mould infections and patulin content in Conference pears when applied 24-48 h after pathogen inoculation. SIGNIFICANCE AND IMPACT OF THE STUDY: trans-2-Hexenal could be a natural alternative to fungicides in the control of P. expansum infections. Further work is needed to study the methods and conditions avoiding the persistence of off-odours and off-flavours in pears after their exposure to trans-2-hexenal vapours. PMID- 16696667 TI - Microbial community responses associated with the development of oomycete plant pathogens on tomato roots in soilless growing systems. AB - AIMS: To determine the spread of different oomycete pathogens in hydroponic, soilless tomato growing systems and their impact on established microbial communities, as baseline studies prior to future introduction of microbial inoculants for disease suppression. METHODS AND RESULTS: The oomycete pathogens, Pythium group F, Pythium aphanidermatum and Phytophthora cryptogea, were introduced into small-scale recirculating tomato growing systems containing rockwool 6 weeks after set-up when roots were well-established. Two weeks later, half of the systems were switched over to run-to-waste. Pythium aphanidermatum spreads the fastest, Pythium group F the slowest and Ph. cryptogea was intermediate in its spread. The switch to run-to-waste had no effect on pathogen recovery. Microbial communities, monitored by dilution plating, were well established at the first sampling, 6 weeks after set-up and although differences in community levels were found between experiments, changes during any one experiment were small, generally less than 1 log10 CFU g(-1) for bacteria. Pathogen introduction increased microbial community levels in roots but the switch to run-to-waste had no effect. Analysis of bacterial communities through amplification of a fragment of the 16S rRNA gene and DGGE profiling showed that different communities were established within each pathogen experiment and that different communities were established on roots, rockwool and in nutrient solutions. However, no significant changes in microbial profiles were found over time in any experiment. CONCLUSIONS: In these systems, the microbial communities were well-established 6 weeks after set-up and were resistant to biological and physical perturbation. SIGNIFICANCE AND IMPACT OF THE STUDY: The implication for microbial inoculation of such systems for disease suppression is that the micro organisms would either have to be introduced very early during the set-up of the system or be able to replace an established but variable community. PMID- 16696668 TI - Interactions between Brettanomyces bruxellensis and other yeast species during the initial stages of winemaking. AB - AIMS: Wine is the product of complex interactions between yeasts and bacteria in grape must. Amongst yeast populations, two groups can be distinguished. The first, named non-Saccharomyces (NS), colonizes, with many other micro-organisms, the surface of grape berries. In the past, NS yeasts were primarily considered as spoilage micro-organisms. However, recent studies have established a positive contribution of certain NS yeasts to wine quality. Amongst the group of NS yeasts, Brettanomyces bruxellensis, which is not prevalent on wine grapes, plays an important part in the evolution of wine aroma. Some of their secondary metabolites, namely volatile phenols, are responsible for wine spoilage. The other group contributing to wine aroma, which is also the main agent of alcoholic fermentation (AF), is composed of Saccharomyces species. The fermenting must is a complex microbial ecosystem where numerous yeast strains grow and die according to their adaptation to the medium. Yeast-yeast interactions occur during winemaking right from the onset of AF. The aim of this study was to describe the interactions between B. bruxellensis, other NS and Saccharomyces cerevisiae during laboratory and practical scale winemaking. METHODS AND RESULTS: Molecular methods such as internal transcribed spacer-restriction fragment length polymorphism and polymerase chain reaction and denaturing gradient gel electrophoresis were used in laboratory scale experiments and cellar observations. The influence of different oenological practices, like the level of sulphiting at harvest time, cold maceration preceding AF, addition of commercial active dry yeasts on B. bruxellensis and other yeast interactions and their evolution during the initial stages of winemaking have been studied. Brettanomyces bruxellensis was the most adapted NS yeast at the beginning of AF, and towards the end of AF it appeared to be more resistant than S. cerevisiae to the conditions of increased alcohol and sugar limitation. CONCLUSIONS: Among all NS yeast species, B. bruxellensis is better adapted than other wild yeasts to resist in must and during AF. Moreover, B. bruxellensis appeared to be more tolerant to ethanol stress than S. cerevisiae and after AF B. bruxellensis was the main yeast species in wine. SIGNIFICANCE AND IMPACT OF THE STUDY: Brettanomyces bruxellensis interacts with other yeast species and adapts to the wine medium as the dominant yeast species at the end of AF. Contamination of B. bruxellensis might take place at the beginning of malolactic fermentation, which is a critical stage in winemaking. PMID- 16696669 TI - Portable continuous flow centrifugation and method 1623 for monitoring of waterborne protozoa from large volumes of various water matrices. AB - AIMS: The aims of this study were to validate a portable continuous flow centrifuge (PCFC) as an alternative concentration step of US-EPA Method 1623 and to demonstrate it's efficacy for recovery of low numbers of protozoa from large volumes of various water matrices. METHODS AND RESULTS: Recoveries of Cryptosporidium parvum oocysts, Giardia intestinalis cysts and Encephalitozoon intestinalis spores spiked into 10-1000 l volumes of various water matrices were evaluated during in-house and collaborative trials. Spiked protozoa were either approved standards or diluted stock samples enumerated according to USEPA Method 1623. Cryptosporidium recoveries exceeded method 1623 criteria and substantially high recoveries were observed for Giardia and E. intestinalis. CONCLUSIONS: Portable continuous flow centrifuge methodology exceeded method 1623 acceptance criteria for Cryptosporidium and could be easily adopted for other protozoa. SIGNIFICANCE AND IMPACT OF THE STUDY: The PCFC could be adopted as an alternative user-friendly concentration method for Cryptosporidium and for monitoring of large volumes of source and tap water for accidental or deliberate contamination with protozoa and potentially with other enteric pathogens. It is anticipated that PCFC would also be equal or superior to filtration for protozoa monitoring in wastewater and effluents. PMID- 16696670 TI - Molecular diversity of rhizobia nodulating the invasive legume Cytisus scoparius in Australia. AB - AIMS: To contribute to the understanding of Cytisus scoparius success at invading and establishing itself in Australia. METHODS AND RESULTS: Root-nodule bacteria isolated from C. scoparius, growing on five different sites and originally introduced to Australia, were compared with isolates from indigenous plants growing in France and isolates from native legumes growing on the same Australian sites as C. scoparius. Small-subunit rDNA from 251 isolates were analysed by PCR RFLP and representatives from different genospecies were selected for sequencing. Phylogenetic analyses revealed a great diversity of lineages belonging to Bradyrhizobium, with one genospecies being specific for Cytisus both in Australia and in France, Rhizobium and Mesorhizobium and one falling outside the described genera of legume-nodulating bacteria. Principal component analysis showed that the Cytisus Australian rhizobial communities are more similar to each other than to their co-occurring native partners. CONCLUSIONS: Early established rhizobial symbionts may have an increased probability to contribute inoculum for the development of further nodules. SIGNIFICANCE AND IMPACT OF THE STUDY: This is a first report comparing rhizobia nodulating C. scoparius in its native and exotic environments. Cytisus scoparius symbionts were identified outside the Bradyrhizobium genus and a new lineage of legume-nodulating bacteria was identified. PMID- 16696671 TI - Isolation and characterization of Legionella spp. and Pseudomonas spp. from greenhouse misting systems. AB - AIMS: Greenhouse misting systems used for watering plants produce fine aerosols. They are a possible cause for bacterial infections. This study investigates the colonization of greenhouse misting systems with Legionella spp. and Pseudomonas spp. and evaluates a possible health hazard. METHODS AND RESULTS: Between June and September 2003, a total of 80 water samples were collected in 20 different greenhouse systems in Germany, each tested on two different occasions. Each time, water was drawn at a central tap and at the outlet of spray nozzles. Sampled greenhouses were used to cultivate various plants and trees for commercial, recreational or scientific reasons, some of them in tropical conditions. Legionella spp. were detected in 10% of the systems (two systems), but only in low numbers. On the contrary, Pseudomonas spp. were recovered from 70% of the greenhouse watering systems (14 systems), occasionally at counts greater than 10,000 CFU per 100 ml. A random amplified polymorphic DNA polymerase chain reaction typing method was used to demonstrate that each colonized greenhouse had one or several individual strains of Legionella and Pseudomonas that could not be detected in any other system. CONCLUSIONS: This study demonstrates that aerosolizing greenhouse watering systems may be contaminated with Legionella or Pseudomonas which under certain circumstances could become a potential source of infection for workers and visitors. SIGNIFICANCE AND IMPACT OF THE STUDY: The study results indicate that greenhouse misting systems should be included in Legionella and Pseudomonas monitoring and control programs. PMID- 16696672 TI - A lacticin 3147 enriched food ingredient reduces Streptococcus mutans isolated from the human oral cavity in saliva. AB - AIMS: To isolate and characterise Streptococcus mutans from Irish saliva samples and to assess their sensitivity to a food-grade preparation of the lantibiotic, lacticin 3147, produced by Lactococcus lactis DPC3147. METHODS AND RESULTS: Saliva samples collected from children with varying oral health status were screened on Mitis Salivarius agar for the presence of pathogenic streptococci. Following selective plating, 16S rDNA sequencing and Pulsed Field Gel Electrophoresis (PFGE), 15 distinct strains of Strep. mutans were identified. These were grouped according to their relative sensitivity to lacticin 3147 which ranged from 0.78 to 6.25%; relative to a sensitive indicator strain, Lactococcus lactis ssp. lactis HP. Inhibition of indicator Strep. mutans strains from sensitive, intermediate and tolerant groupings were assessed in microtitre plate assays with increasing concentrations of lacticin 3147. The concentration of lacticin 3147 required to give 50% growth inhibition correlated with their relative sensitivities (as assayed by well diffusion methodology) and ranged from 1280 to 5120 AU ml(-1). Concentrated preparations of lacticin 3147 caused a rapid killing of Strep. mutans strains in broth. Moreover, in human saliva deliberately spiked with Strep. mutans, the pathogen was eliminated (initial inoculum of 10(5)) in the presence of 40,000 AU ml(-1) of lacticin 3147. Furthermore, a food grade lacticin 3147 spray dried powder ingredient was assessed for the inhibition of Strep. mutans in human saliva, spiked with a strain of intermediate sensitivity, resulting in up to a 4-log reduction in counts after 20 min. CONCLUSION: A food grade preparation of lacticin 3147 was effective in the inhibition of oral Strep. mutans. SIGNIFICANCE AND IMPACT OF THE STUDY: The inhibition of oral streptococci by food grade preparations of lacticin 3147 may offer novel opportunities for the development of lacticin 3147 as an anti cariogenic agent particularly in the area of functional foods for the improvement of oral health. PMID- 16696673 TI - Production of indolic compounds by rumen bacteria isolated from grazing ruminants. AB - AIM: To screen rumen bacterial cultures and fresh ruminal isolates for indole and skatole production. METHODS AND RESULTS: Culture collection strains and fresh bacterial isolates from rumen contents of sheep and dairy cows were screened for the production of indolic compounds. Clostridium aminophilum FT, Peptostreptococcus ssp. S1, Fusobacterium necrophorum D4 produced indole and Clostridium sticklandii SR produced indoleacetic acid. Fresh isolates from sheep (TrE9262 and TrE7262) and dairy cows (152R-1a, 152R-1b, 152R-3 and 152R-4) produced indole, indolepropionic acid, tryptophol and skatole from the fermentation of tryptophan and indoleacetic acid. Glucose altered the indolic compounds produced in some, but not all, isolates. TrE7262 and 152R-4 were identified as Clostridium sporogenes and 152R-1b as a new Cl. aminophilum strain. Isolates TrE9262, 152R-1a and 152R-3 were not closely related to any described species but belong to Megasphaera, Prevotella and Actinomyces genera, respectively. CONCLUSIONS: Rumen bacteria that produced a range of indolic compounds were identified. Some isolates are distinct from the previously described bacteria and may represent novel species. SIGNIFICANCE AND IMPACT OF THE STUDY: These observations will contribute to understanding skatole and indole formation in the rumen and will lead to methods that control the formation of indolic compounds in pasture-grazed ruminants. PMID- 16696674 TI - Comparison of 16S rRNA sequencing with conventional and commercial phenotypic techniques for identification of enterococci from the marine environment. AB - AIMS: To compare accuracy of genus and species level identification of presumptive enterococci isolates from the marine environment using conventional biochemical testing, four commercial identification systems and 16S rRNA sequence analysis. METHODS AND RESULTS: Ninety-seven environmental bacterial isolates identified as presumptive enterococci on mEI media were tested using conventional and Enterococcus genus screen biochemical tests, four commercial testing systems and 16S rRNA sequencing. Conventional and Enterococcus genus screen biochemical testing, 16S rRNA sequencing and two commercial test systems achieved an accuracy of > or = 94% for Enterococcus genus confirmation. Conventional biochemical testing and 16S rRNA sequencing achieved an accuracy of > or = 90% for species level identification. CONCLUSIONS: For confirmation of Enterococcus genus from mEI media, conventional or genus screen biochemical testing, 16S rRNA sequencing and the four commercial systems were correct 79-100% of the time. For speciation to an accuracy of 90% or better, either conventional biochemical testing or 16S rRNA sequencing is required. SIGNIFICANCE AND IMPACT OF THE STUDY: Accurate identification of presumptive environmental Enterococcus isolates to genus and species level is an integral part of laboratory quality assurance and further characterization of Enterococcus species from pollution incidents. This investigation determines the ability of six different methods to correctly identify environmental isolates. PMID- 16696675 TI - horC confers beer-spoilage ability on hop-sensitive Lactobacillus brevis ABBC45cc. AB - AIMS: To determine whether horC confers beer-spoilage ability and to evaluate the validity of horC as a trans-species genetic marker for differentiating the beer spoilage ability of lactic acid bacteria (LAB). METHODS AND RESULTS: Hop sensitive Lactobacillus brevis ABBC45cc was transformed with an expression plasmid, pHYchorBC, containing putative multidrug resistance gene horC and its putative regulator horB, and the transformant was designated as ABBC45cc/pHYchorBC. As a control, ABBC45cc was transformed with pHYchorB that contains horB, and the transformed strain was designated as ABBC45cc/pHYchorB. As a result of beer-spoilage assay of these transformants, ABBC45cc/pHYchorBC exhibited beer-spoilage ability, whereas ABBC45cc/pHYchorB did not. Furthermore ABBC45cc/pHYchorBC showed higher hop resistance than ABBC45cc/pHYchorB, accounting for the differences in beer-spoilage ability observed between the two transformants. ABBC45cc/pHYchorBC also exhibited higher resistance to various structurally unrelated drugs, compared with ABBC45cc/pHYchorB. CONCLUSIONS: horC was shown to confer hop resistance and beer-spoilage ability on ABBC45cc by presumably encoding a multidrug transporter. SIGNIFICANCE AND IMPACT OF THE STUDY: The finding that horC plays an important role in hop resistance and beer spoilage ability supports the validity of horC as a trans-species genetic marker for differentiating the beer-spoilage ability of LAB. PMID- 16696676 TI - Dynamic modeling of Listeria monocytogenes growth in pasteurized milk. AB - AIMS: The development and validation of a dynamic model for predicting Listeria monocytogenes growth in pasteurized milk stored at both static and dynamic temperature conditions. METHODS AND RESULTS: Growth of inoculated L. monocytogenes in a commercial pasteurized whole milk product was monitored at various isothermal conditions from 1.5 to 16 degrees C. The kinetic parameters of the pathogen were modelled as a function of temperature using a square root type model, which was further validated using data from 92 published growth curves from eight different milk products. Compared to four published models for L. monocytogenes growth, the model developed in this study performed better, with a per cent discrepancy and bias of 49.1 and -1.01%, respectively. The performance of the model in predicting growth at dynamic temperature conditions was evaluated at four different fluctuating temperature scenarios with periodic temperature changes from -2 to 16 degrees C. The prediction of growth at dynamic storage temperature was based on the square root model in conjunction with the differential equations of the Baranyi and Roberts model, which were numerically integrated with respect to time. The per cent relative errors between the observed and the predicted growth of L. monocytogenes were less than 10% for all temperature scenarios tested. CONCLUSIONS: Available models from experiments conducted in laboratory media may result in significant overestimation of L. monocytogenes growth in pasteurized milk because they do not take into account factors such as milk composition (e.g. natural antimicrobial compounds present in milk) and the interactions of the pathogen with the natural microflora. The product-targeted model developed in the present study showed a high performance in predicting growth of L. monocytogenes in pasteurized milk under both static and dynamic temperature conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Temperature fluctuations often occur during the transportation and storage of pasteurized milk. A high performance, dynamic model for the growth of L. monocytogenes can be a useful tool for effective management and optimization of product safety and can lead to more realistic estimations of pasteurized-milk related safety risks. PMID- 16696677 TI - Growth studies of plasmid bearing and plasmid cured Yersinia enterocolitica GER O:3 in the presence of cefsulodin, irgasan and novobiocin at 25 and 37 degrees C. AB - AIM: In this study, the growth characteristics of Yersinia enterocolitica biotype 4, GER O:3 plasmid bearing (P+) and plasmid cured (P-) strain types were evaluated in brain heart infusion broth supplemented with cefsulodin, irgasan, and novobiocin alone or in combination. METHODS AND RESULTS: Growth curves were obtained for the two strain types in broth supplemented with selective agents at 25 or 37 degrees C for 32 h to obtain data on the lag phase durations and growth rates of the strains. Generally, the lag times and growth rates of the P+ and P- strains were similar for cultures incubated at 25 degrees C regardless of the selective agent added and where plasmid replication and expression were not under any significant burden. However, where the lag times and growth rates of the strains were examined at 37 degrees C, significant differences were observed in the lag phase durations of the plasmid bearing strain type compared the plasmid cured strain, an effect that was due to the burden of the plasmid and the influence of selective agents. Generally, when two or more agents were present, lag phase durations were longer for the plasmid bearing strain. Some exceptions noted where in the presence of irgasan or full selective agent (CIN) the opposite case was observed. When growth rates were compared, the plasmidless strain type was typically faster than the plasmid bearing strain in the presence of most selective agents at 37 degrees C and the growth rates of both strain types at 25 degrees C were similar where the temperature appeared to negate the effects of plasmid. CONCLUSIONS: The data obtained in these studies suggest that selective agents (in particular irgasan) and incubation temperature play a significant role in influencing the growth characteristics of plasmid bearing and plasmid cured strains of Y. enterocolitica. SIGNIFICANCE AND IMPACT OF THE STUDY: This data presented in this study has significant implications for enrichment methods used in the detection or recovery of plasmid bearing Y. enterocolitica strains from food, environmental or clinical samples. PMID- 16696678 TI - Conversion of Lactococcus lactis cell envelope proteinase specificity by partial allele exchange. AB - AIMS: To determine whether conversion of lactocepin substrate binding regions by gene replacement can alter lactocepin specificity in Lactococcus lactis starter bacteria without affecting other important strain properties. METHODS AND RESULTS: We utilized two-step gene replacement to convert substrate-binding determinants in the L. lactis prtP genes encoding group h (bitter) lactocepin in two industrial strains into the corresponding group b (nonbitter) variant. Analysis of lactocepin activity toward alpha(s1)-casein (f 1-23) by reversed phase high-pressure liquid chromatography demonstrated enzyme specificity among isogenic derivatives had been altered in a manner that was consistent with predicted amino acid substitutions in substrate binding regions. Milk acidification properties of some mutants were not statistically different (P > 0.05) from wild-type parent strains, and strain propensity for autolysis was also not significantly (P > 0.05) changed. CONCLUSIONS: Conversion of lactocepin substrate binding regions by allele exchange can effectively alter lactocepin specificity in industrial strains of L. lactis without significantly affecting other important strain properties. SIGNIFICANCE AND IMPACT OF THE STUDY: Methodology outlined in this study can be used to alter lactocepin specificity in commercial starter cultures with a propensity for bitter flavour defect, and prtP derivatives developed by this approach should be suitable for commercial application. PMID- 16696679 TI - Detection and characterization of hepatitis A virus in water samples in Thailand. AB - AIMS: Outbreaks of hepatitis A in Thailand have been reported continuely and associated with water supply. However, the genetic analysis of hepatitis A virus (HAV) in water is limited. This study described the application of virus concentration method and reverse transcriptase-nested polymerase chain reaction (RT-nested PCR) to detect HAV RNA and analyse the genetic sequence of the virus in environmental water samples. METHODS AND RESULTS: The HAV from water samples was concentrated by using a developed virus concentration method (adsorption elution and subsequent speedVac reconcentration) and the viral RNA was detected by RT-nested PCR followed by sequencing of the amplified DNA products. Detection limit of HAV determined by the RT-nested PCR was 1.29 radioimmunofocus assay (RIFA) units ml(-1). The DNA band appeared at 183 basepairs. No cross-reactivity was observed in the presence of other enteric viruses (poliovirus and rotavirus). A total of 180 water samples were collected, concentrated, and detected for HAV. The HAV was found in 6/40 (15%) of water samples collected from a swamp and 3/30 (10%) collected from a canal. Ten river samples and 100 tap water samples stored in containers for drinking and domestic uses were negative for HAV. In sequence analysis of the DNA products and alignment with the HAV sequence deposited in the GenBank, six water samples showed the nucleotide sequence associated with HAV. The 120 nucleotides in the N-terminal VP1 region obtained from two swamp samples showed 95 and 96.7% identity to HAV genotype IA. In nearly all water samples where HAV was present bacterial indicators (faecal coliforms and Escherichia coli) were found for faecal contamination. CONCLUSIONS: A coupled virus concentration method and RT-nested PCR was successfully applied to examine HAV in water samples collected from various sources. DNA sequencing of nested PCR products showed the genotype IA associated with HAV that is predominate in Thailand. SIGNIFICANCE AND IMPACT OF THE STUDY: This research is the first study of genetic sequence of HAV in water samples in Thailand. The presence of naturally occurring HAV might pose a potential health risk for people. PMID- 16696680 TI - Antagonistic activity of probiotic lactobacilli and bifidobacteria against entero and uropathogens. AB - AIM: To develop in vitro assays for comparing the antagonistic properties and anti-oxidative activity of probiotic Lactobacillus and Bifidobacterium strains against various entero- and urinary pathogens. METHODS AND RESULTS: The antagonistic activity of five probiotic lactobacilli (Lactobacillus rhamnosus GG, Lactobacillus fermentum ME-3, Lactobacillus acidophilus La5, Lactobacillus plantarum 299v and Lactobacillus paracasei 8700:2) and two bifidobacteria (Bifidobacterium lactis Bb12, Bifidobacterium longum 46) against six target pathogens was estimated using different assays (solid and liquid media, anaerobic and microaerobic cultivation) and ranked (low, intermediate and high). Bacterial fermentation products were determined by gas chromatography, and the total anti oxidative activity of probiotics was measured using linolenic acid test. Pyelonephritic Escherichia coli was highly suppressed by GG and both bifidobacteria strains. Lactobacilli strains 8700:2, 299v and ME-3 were the most effective against Salmonella enterica ssp. enterica in microaerobic while ME-3 and both bifidobacteria expressed high activity against Shigella sonnei in anaerobic milieu. Lact. paracasei, Lact. rhamnosus and Lact. plantarum strains showed intermediate antagonistic activity against Helicobacter pylori under microaerobic conditions on solid media. The highest anti-oxidative activity was characteristic for Lact. fermentum ME-3 (P < 0.05). No efficient antagonist against Clostridium difficile was found. The positive correlations between the pH, lactic acid production and anti-microbial activity for all tested probiotics were assessed. CONCLUSIONS: Developed experimental assays enable to compare the anti-microbial and -oxidative activity of Lactobacillus and/or Bifidobacterium probiotics, which have been claimed to possess the ability of suppressing the growth of various enteric and urinary pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: Screening Lactobacillus and Bifidobacterium sp. strains according to their activity in various environmental conditions could precede the clinical efficacy studies for adjunct treatment with probiotics in cure of different gastrointestinal and urinary tract infections. PMID- 16696681 TI - Antifungal activity of Juniperus essential oils against dermatophyte, Aspergillus and Candida strains. AB - AIMS: The increasing resistance to antifungal compounds and the reduced number of available drugs led us to search therapeutic alternatives among aromatic plants and their essential oils, empirically used by antifungal proprieties. In this work the authors report on the antifungal activity of Juniperus essential oils (Juniperus communis ssp. alpina, J. oxycedrus ssp. oxycedrus and J. turbinata). METHODS AND RESULTS: Antifungal activity was evaluated by determination of MIC and MLC values, using a macrodilution method (NCCLS protocols), on clinical and type strains of Candida, Aspergillus and dermatophytes. The composition of the oils was ascertained by GC and GC/MS analysis. All essential oils inhibited test dermatophyte strains. The oil from leaves of J. oxycedrus ssp. oxycedrus is the most active, with MIC and MLC values ranging from 0.08-0.16 microl ml(-1) to 0.08 0.32 microl ml(-1), respectively. This oil is mainly composed of alpha-pinene (65.5%) and delta-3-carene (5.7%). CONCLUSIONS: J. oxycedrus ssp. oxycedrus leaf oil proved to be an emergent alternative as antifungal agent against dermatophyte strains. delta-3-Carene, was shown to be a fundamental compound for this activity. SIGNIFICANCE AND IMPACT OF THE STUDY: Results support that essential oils or some of their constituents may be useful in the clinical management of fungal infections, justifying future clinical trials to validate their use as therapeutic alternatives for dermatophytosis. PMID- 16696682 TI - Comparison of culture media and chairside assays for enumerating mutans streptococci. AB - AIM: This study compared several traditional culture-based media and chairside cultural assays for ability to recover mutans streptococci (MS) from pure cultures and from saliva samples. METHODS AND RESULTS: When pure cultures were used with traditional culture-based media, mitis-salivarius bacitracin (MSB) agar demonstrated less support for bacterial recovery than trypticase-yeast extract cysteine sucrose-bacitracin (TYCSB) agar and the modified medium of Ritz (HLR-S). One species of MS, Streptococcus ferus (c), was not recovered on MSB medium. Chairside cultural tests displayed considerable disparity between tests in recovering bacteria from pure cultures. On the glass adherence assay (Mucount), S. ferus was not detected and Streptococcus criceti was not detected on the dipslide assay (Cariescreen SM) or on the plastic adherence assay (Dentocult SM Strip mutans). The frequency of isolation of pure strains of bacteria other than MS was common. From saliva samples, the frequency of isolation of MS on HLR-S and TYCSB media and the glass adherence assay was 91-97%. The frequency of isolation on MSB medium and on the dip-slide and plastic adherence assays was significantly decreased (37, 47 and 69%, respectively). Recovery scores varied considerably among the culture methods studied and tended to be highest on the HLR-S medium and on the glass adherence assay. CONCLUSIONS: Growth and recovery profiles of pure bacterial cultures and of saliva samples for the MS varied according to different media. SIGNIFICANCE AND IMPACT OF THE STUDY: Caution should be exercised in comparing results between studies that employ different cultural methods for MS enumeration. PMID- 16696683 TI - A cost effective fermentative production of succinic acid from cane molasses and corn steep liquor by Escherichia coli. AB - AIM: Development and optimization of an efficient and inexpensive medium for succinic acid production by Escherichia coli under anaerobic conditions. METHODS AND RESULTS: Initially, 0.8 gl(-1) of succinic acid was produced in 60 h in 300 ml medium. On optimization, glucose and peptone were replaced by cane molasses and corn steep liquor. Three hundred ml of this medium was inoculated with 4% (v/v) of seed inoculum, incubated at 39 degrees C for 72 h, resulted in 7.1 gl( 1) of succinic acid in 36 h. Scale up in a 10-l fermentor under conditions of controlled pH and continuous CO2 supply in this medium resulted in 17 gl(-1) of succinic acid in 30 h. CONCLUSIONS: A ninefold increase in succinic acid production was obtained in 500-ml anaerobic bottles with optimized medium having cane molasses and corn steep liquor as against initial medium containing glucose and peptone. However, a subsequent scale up in a 10-l fermentor resulted in a 2.5 fold increase in succinic acid production as against optimized medium used in 500 ml anaerobic bottles. SIGNIFICANCE AND IMPACT OF THE STUDY: Succinic acid production was enhanced in medium consisting of inexpensive carbon and nitrogen sources in a shorter span of time. PMID- 16696684 TI - Characterization of dominant microbiota of a Ghanaian fermented milk product, nyarmie, by culture- and nonculture-based methods. AB - AIMS: To characterize the predominant micro-organisms in a Ghanaian traditional fermented dairy product, nyarmie, made from cows' milk, using both culture- and nonculture-based methods. METHODS AND RESULTS: Samples of nyarmie were analysed from three production sites in Accra, by determining the counts on selective culture media. The microbial diversity occurring in nyarmie was also evaluated by 16S/18S ribosomal DNA PCR amplification and denaturing gradient gel electrophoresis. Results showed that nyarmie contained lactococci and lactobacilli in the range of 10(8) and 10(10) CFU ml(-1), respectively, and yeasts at around 10(7) CFU ml(-1). The pH ranged between 3.49 and 4.25. The predominant lactic acid bacteria (LAB) in nyarmie were Leuconostocmesenteroides ssp. mesenteroides, Streptococcus thermophilus, Lactobacillus delbrueckii ssp. bulgaricus, Lact.helveticus, Lact. delbrueckii ssp. lactis and Lactococcus lactis, while Saccharomyces cerevisiae was the predominant yeast species. Lactobacillus delbrueckii ssp. delbrueckii was not detected by cultivation but its predominance was revealed by PCR-DGGE analysis. CONCLUSIONS: The flora in products from different producers varied in the LAB composition present and may result in variations in product quality. SIGNIFICANCE AND IMPACT OF THE STUDY: Development and use of starter cultures for nyarmie may be beneficial in improving the consistency of product quality. PMID- 16696685 TI - Glyceraldehyde-3-phosphate dehydrogenase regulation in Lactococcus lactis ssp. cremoris MG1363 or relA mutant at low pH. AB - AIMS: To analyse the phenotype of a relA acid-resistant mutant of Lactococcus lactis ssp. cremoris MG1363, and to compare the glyceraldehyde-3-phosphate dehydrogenase regulation in both strains. METHODS AND RESULTS: Lactococcus lactis ssp. cremoris MG1363 and the relA mutant affected in the (p)ppGpp synthetase were grown in a series of batch-mode fermentation at different pH-regulated conditions with glucose as carbon substrate. All the determinants of the glyceraldehyde-3 phosphate dehydrogenase (GAPDH) regulation were quantified. In L. lactis MG1363, the GAPDH was strongly inhibited in vitro by decreased pH values, but this inhibition was totally compensated in vivo by the lower NADH/NAD+ ratio and more efficiently by the important increase in the intracellular amount of GAPDH. In contrast to the wild type, GAPDH activity of the relA strain was not increased when grown at low pH but the level of GAPDH remained constitutively high. However, pH homeostasis was not improved in the relA mutant and it grew slower and exhibited a lower glycolytic flux than the wild-type strain at low pH. CONCLUSIONS: Despite a better resistance to acid stress, the increased survival in L. lactis relA mutant at low pH was not related with an improved pH homeostasis but was associated with a diminished capacity to maintain a high flux through glycolysis. SIGNIFICANCE AND IMPACT OF THE STUDY: The phenotype of a strong acid-resistant L. lactis strain was established in acid conditions and some key metabolic parameters compared with the wild type. This analysis led to the conclusion that growth and survival seem to be antinomic parameters, since improving one of them leads to a decrease in the other one. PMID- 16696686 TI - FK506, a secondary metabolite produced by Streptomyces, presents a novel antiviral activity against Orthopoxvirus infection in cell culture. AB - AIMS: To investigate the antiviral potential of the macrolide FK506, produced by Streptomyces tsukubaensis, against Orthopoxvirus infection in cell culture, and determine the replicative stage of viral cycle affected by the treatment. METHODS AND RESULTS: Cell lines were infected with different Orthopoxviruses and treated with FK506. The macrolide inhibited the replication of the prototypic Orthopoxvirus, vaccinia virus strain WR, with an IC50 of 12.05 micromol l(-1). Progeny production of other Orthopoxviruses was also inhibited by FK506 at noncytotoxic concentrations, as evaluated by the neutral-red uptake assay and metabolic labelling of cellular proteins. By Western blot assay, we detected a severe inhibition (approximately 87.6% +/- 2.78%) of VV strain WR post replicative protein synthesis. A similar reduction of virus DNA accumulation, as observed by slot-blot assay, probably accounts for the subsequent inhibition of virus late proteins. CONCLUSIONS: The macrolide FK506, isolated from S. tsukubaensis, presents a novel anti-poxvirus activity, probably targeting the stage of DNA replication during Orthopoxvirus infection. SIGNIFICANCE AND IMPACT OF THE STUDY: The secondary metabolite FK506, isolated from the culture filtrate of S. tsukubaensis, shows a pleiotropic range of activities, and might be a valuable tool as a lead structure in the generation of non-immunosuppressant analogues with strong anti-poxvirus activity. PMID- 16696687 TI - Detection of fish nocardiosis by loop-mediated isothermal amplification. AB - AIMS: Loop-mediated isothermal amplification (LAMP) is a novel method that amplifies DNA with high specificity and rapidity under isothermal conditions. In this study, using the LAMP method, a protocol for detecting Nocardia seriolae which is a causative agent of fish nocardiosis, was designed. METHODS AND RESULTS: A set of four primers, two inner and two outer, were designed based on the sequence of the 16S-23S ribosomal RNA internal transcribed spacer region of N. seriolae. Time and temperature conditions for detection of N. seriolae were optimized for 60 min at 65 degrees C. Other fish pathogen was not amplified by this LAMP system. The detection of N. seriola using LAMP was found to be more sensitive than that by polymerase chain reaction. CONCLUSIONS: LAMP is a highly sensitive and rapid diagnostic procedure for detection of N. seriolae. SIGNIFICANCE AND IMPACT OF THE STUDY: LAMP is a useful diagnostic method for fish nocardiosis. PMID- 16696688 TI - Production of organic acids from fermentation of mannitol, fructooligosaccharides and inulin by a cholesterol removing Lactobacillus acidophilus strain. PMID- 16696690 TI - The application of geographic information systems and spatial analysis to assess dumped and subsequently scattered human remains. AB - This study utilizes geographic information systems (GIS) and spatial analysis (SA) technology to address the problems associated with prediction of location and effective recovery of dumped and scattered human remains in Louisiana. The goals are to determine if a selective bias exists in Louisiana as to where and when human remains are dumped and to assess whether or not geographically specific patterns exist in the dispersal of human remains. We hypothesized that a positive relationship exists between postmortem interval (PMI) and dispersal distance, and that there are negative relationships between PMI and dispersal direction and between dispersal direction and distance. Our results indicate that, in Louisiana, remains are more often dumped in rural areas away from a structure, and are found within (1/4) mile of the nearest road. For Louisiana, no seasonal bias was found in the analysis of when remains are dumped. Furthermore, with the exception of the relationship between PMI and the shortest distance remains were dispersed, no geographically specific patterns were detected in the analyses of dispersal distance, dispersal direction, and PMI. PMID- 16696691 TI - Taphonomy of child-sized remains: a study of scattering and scavenging in Virginia, USA. AB - Child-sized pig carcasses (Sus scrofa) were placed in surface deposit and buried scenarios in a wooded area of Virginia from May 1998 through December 2000, to examine the taphonomic effects of decompositional changes, predator scavenging, and the extent of remains scattering. Changes were observed through on-site examination, charting of remains, and recorded video imaging. Analysis of data revealed that utilization of corpses as food sources by vertebrates was dependent upon invertebrate colonization. Vertebrates avoided feeding on the corpses while invertebrate colonization was active, and would feed before invertebrates successfully colonized a corpse, or would wait until the invertebrate populations migrated away from the corpse. Among vertebrates, there was no apparent succession order for the animals utilizing the remains as a food source. Different vertebrates would feed at different times based upon diurnal or nocturnal predilection. Analysis noted an accidental cooperative relationship between the invertebrates and vertebrates scavenging on the corpses. Certain vertebrates gained access to the internal tissues by utilizing openings in the corpses caused by invertebrate and other vertebrate scavenging. Alternately, carrion-frequenting insects were afforded access to previously inaccessible colonization sites as a result of scavenging vertebrate activities. PMID- 16696692 TI - Sacroiliac joint bridging: simple and reliable criteria for sexing the skeleton. AB - Vital to the study of past populations and an important phase in executing a forensic anthropological examination is the determination of skeleton sex. The aim of this study is to present an easy and reliable criterion for sexing the skeleton. The ilium and sacrum of 2845 skeletons were examined for new bone formation and for bridging/fusion in/across the sacroiliac joint. Sacroiliac joint bridging (SIB) was present in 12.27% of the males and 1.83% of the females. In 97% of the males bridging was extra-articular, whereas in all females bridging was intra-articular. In addition, computed tomograghy images of 81 in vivo individuals were examined for the same phenomenon. SIB was present in 34.2% of the males and 4.6% of the females. Bony spurs present on the ilium for a preliminary partial or full extra-articular bridging of the sacroiliac joint indicate a male skeleton. SIB presents an easy technique for sexing skeletons (especially in elderly individuals where the phenomenon of SIB becomes very common), as no prior knowledge, training, or equipment is required to apply the criterion. PMID- 16696693 TI - Analysis of artificial radiocarbon in different skeletal and dental tissue types to evaluate date of death. AB - Radiocarbon dating, with special reference to the modern bomb-curve, can provide useful information to elucidate the date of death of skeletonized human remains. Interpretation can be enhanced with analysis of different types of tissues within a single skeleton because of the known variability of formation times and remodeling rates. Analysis of radiocarbon content of teeth, especially the enamel in tooth crowns, provides information about the date of formation in the childhood years and in consideration of the known timing of tooth formation can be used to estimate the birth date after 1950 ad. Radiocarbon analysis of modern cortical and trabecular bone samples from the same skeleton may allow proper placement on the pre-1963 or post-1963 sides of the bomb-curve as most trabecular bone generally undergoes more rapid remodeling than does most cortical bone. Pre 1963 bone formation would produce higher radiocarbon values for most trabecular bone than for most cortical bone. This relationship is reversed for formation after 1963. Radiocarbon analysis was conducted in this study on dental, cortical, and trabecular bone samples from two adult individuals of known birth (1925 and 1926) and death dates (1995 and 1959). As expected, the dental results correspond to prebomb-curve values reflecting conditions during the childhoods of the individuals. The radiocarbon content of most bone samples reflected the higher modern bomb-curve values. Within the bone sample analyses, the values of the trabecular bone were higher than those of cortical bone and supported the known placement on the pre-1963 side of the bomb-curve. PMID- 16696694 TI - Dietary health does affect histological age assessment: an evaluation of the Stout and Paine (1992) age estimation equation using secondary osteons from the rib. AB - An age at death estimation equation that uses rib histological variables presented by Stout and Paine was used to evaluate a skeletal population of individuals with a known age at death and cause of death from either malnutrition or the niacin deficiency disease pellagra. The sample was comprised of 26 autopsied black South Africans. Histological analysis of mounted thin sections involved the microscopic measurement of cortical area and a count of the number of intact and fragmentary secondary osteons for the entire cross-section of the rib. Rib osteon population density values were then calculated for each case. It was found that this equation under-aged individuals on average by 29.2 years. Overall, secondary osteon size and Haversian canals tended to be larger than expected, while cortical bone area was less when compared with a control population. The implications of these findings are critical given that many of the skeletal remains examined by forensic anthropologists come from marginalized backgrounds, including malnutrition. This research suggests that measurements based on healthy cases may not be useful in an analysis of individuals with poor diet and health. It is argued that new standards for histological age assessment methods need to be created that account for variation in the health status of individuals examined by forensic anthropologists. PMID- 16696695 TI - A soft-tissue calcification: differential diagnosis and pathogenesis. AB - An autopsy of a 72-year-old white male revealed a 30.5 x 5.1 cm vertically aligned heterotopic ossification just deep to a 30.5 cm midline abdominal scar. The ossified mass was determined to be a heterotopic ossification or myostitis ossificans (MO) traumatica resulting from an abdominal surgical incision during life. While, MO traumatica is relatively common accounting for roughly 60-75% of patients with soft-tissue ossification, heterotopic ossification of abdominal incisions are relatively rare and thus infrequently reported. This article details the manifestation of this relatively large heterotopic bone and provides a comprehensive review of the literature and pathogenesis of this unusual ossification. A review of the English literature from 1920 to the present produced only a handful of articles for a total of 185 reported cases. All had bone formed within vertical incisions, usually within 1 year of surgery, and 89% were males. Knowledge of this phenomenon and the variable size at presentation is useful to both the autopsy pathologist and the anthropologist in generating a diagnosis for abnormal calcifications. PMID- 16696696 TI - A forensic case study of a naturally mummified brain from the bushveld of South Africa. AB - The present study reports our observations of a naturally mummified human brain found in the bushveld of South Africa. This case extends the geographic and climatic ranges in which mummified brains have been found, and it represents an additional case where no human activity has led to the mummification. The mummified brain was c. one fifth the size of a normal human brain, while the gyral and sulcal patterns of a typical human brain were clear. CT scanning of the brain revealed that subcortical structures, normally evident in this type of imaging, were not discernable, indicating a slow mummification process. Histological examination of the tissue revealed near complete degradation of the microanatomical structure, with only putative Nissl bodies remaining as identifiable neural microstructures. The specimen appears to have survived several veld fires, as well as a high annual rainfall, and a high relative humidity. It is thought that specific conditions amenable to brain mummification, but not other soft tissues, occurred in the skull of this specimen in the weeks postmortem. PMID- 16696697 TI - A validation study for duct tape end matches. AB - Fracture matches are considered to be the strongest association achievable in forensic examinations of glass, metal, wood, plastic, paint, tape, and other trace evidence. Despite being fairly routine examinations, few publications exist to support their admissibility in court. This study was designed, using duct tape as the fractured medium, to determine the validity and error rate associated with conducting end match (fracture match) examinations on this material. Five test designs, which varied either the source roll of tape or manner of separation (torn or cut) from the roll, were administered to four analysts with instructions to examine the assigned test sets for end matches. If an end match was not identified by the initial analyst, the entire test set was independently evaluated by the remaining three analysts. Results indicated that while tape grade did not hinder end match identification, the manner of separation could affect results. PMID- 16696698 TI - Lead-free primer residues: a qualitative characterization of Winchester WinClean, Remington/UMC LeadLess, Federal BallistiClean, and Speer Lawman CleanFire handgun ammunition. AB - The elemental composition and postfiring residues of several "lead-free" or "nontoxic" centerfire handgun ammunition types currently available to the general public were examined. Offerings from Winchester, Remington/UMC, Federal and Speer were obtained from retail sources in both .45 ACP and 9 mm when possible. A total of 112 postfiring residue samples (SEM pins) were collected at varying distances from the muzzle, at two distances from target and from the shooter's hands. An additional 20 samples were collected by direct ignition of primers. Qualitative determinations were carried out using scanning electron microscopy (SEM)/electron dispersive spectrometry (EDS) analysis. All types tested contained aluminum (Al), silicon (Si), copper (Cu), and zinc (Zn). Most contain traces of sulfur and calcium. Winchester WinClean and Remington LeadLess contained potassium as the principle ingredient. Federal BallistClean contained barium, while Speer Lawman CleanFire contained strontium. In the main, these compared favorably with manufacturers' MSDS publications and patents granted. The characterizations undertaken here will be of use to the forensic electron microscopist as these formulations gain popularity. PMID- 16696699 TI - Metal objects mapping after small charge explosions. A study on AISI 304Cu steel with two different grain sizes. AB - Evidence of exposure of a metal component to a small charge explosion can be detected by observing microstructural modifications; they may be present even if the piece does not show noticeable overall plastic deformations. Particularly, if an austenitic stainless steel (or another metal having a face-centered cubic structure and a low stacking fault energy) is exposed to an explosive shock wave, high-speed deformation induces primarily mechanical twinning, whereas, in nonexplosive events, a lower velocity plastic deformation first induces slip. The occurrence of mechanical twins can be detected even if the surface is damaged or oxidized in successive events. In the present research, optical metallography (OM) and scanning electron microscopy (SEM), and scanning tunneling microscopy (STM) were used to detect microstructural modifications caused on AISI 304Cu steel disks by small-charge explosions. Spherical charges of 54.5 or 109 g TNT equivalent mass were used at explosive-to-target distances from 6.5 to 81.5 cm, achieving peak pressures from 160 to 0.5 MPa. Explosions induced limited or no macro-deformation. Two alloy grain sizes were tested. Surface OM and SEM evidenced partial surface melting, zones with recrystallization phenomena, and intense mechanical twinning, which was also detected by STM and X-ray diffraction. In the samples' interior, only twins were seen, up to some distance from the explosion impinged surface and again, at the shortest charge-to-sample distances, in a thin layer around the reflecting surface. For forensic science locating purposes after explosions, the maximum charge-to-target distance at which the phenomena disappear was singled out for each charge or grain size and related to the critical resolved shear stress for twinning. PMID- 16696700 TI - Infrared spectra of U.S. automobile original finishes. VII. Extended range FT-IR and XRF analyses of inorganic pigments in situ--nickel titanate and chrome titanate. AB - The identification, analysis, and occurrence in U.S. automobile original finishes (1974-1989) of Nickel Titanate (yellow) and Chrome Titanate (yellow-orange) are described in this report. The titanate pigments are based on the rutile (titanium dioxide) structure and there are only minor differences between the infrared absorptions of rutile and the titanates. Titanate pigment absorptions in paint spectra can thus be easily mistaken for those of rutile. Each of the titanates, however, contains two elements in addition to titanium that can serve to distinguish them using elemental analyses. Fourier transform infrared (4000-220 cm(-1)) and X-ray fluorescence instruments were used in combination for the in situ analysis of the titanates. In addition to titanium, nickel, and antimony, the three main detectable elements comprising Nickel Titanate, all of the commercial products of this pigment that were examined also contained impurities of zirconium, niobium, and usually lead. These elements were also detected in most of the monocoats in which Nickel Titanate was identified, as well as in the Chrome Titanate pigments, and the zirconium to niobium ratio was found to exhibit a wide variation. Nickel Titanate is a relatively common pigment that was identified in nearly three dozen U.S. automobile yellow nonmetallic monocoats (1974-1989), while Chrome Titanate appears to have been used in only a few yellow and orange nonmetallic monocoats. The use of the titanate pigments likely increased after this time period as they were replacements for lead chromate pigments (last used in a U.S. automobile original finish in the early 1990s), and are more amenable for use in basecoat/clearcoat finishes than in monocoats. Minor distortions of the infrared absorptions of rutile, anatase, and the titanates obtained using accessories with diamond windows were noted, and their origins are discussed. PMID- 16696701 TI - Environmental survey for four pathogenic bacteria and closely related species using phylogenetic and functional genes. AB - Bacterial species with high DNA sequence similarity to pathogens could affect the specificity of assays designed to detect biological threat agents in environmental samples. The natural presence of four pathogenic bacteria, Bacillus anthracis, Clostridium perfringens, Francisella tularensis, and Yersinia pestis and their closely related species, was determined for a large collection of soil and aerosol samples. Polymerase chain reaction (PCR) and gene sequencing were used using group-specific 16S rRNA primers to identify pathogens and related species, and pathogen-specific virulence genes. Close relatives of B. anthracis (B. cereus group species) were detected in 37% of the soils and 25% of the aerosol samples. The B. anthracis protective antigen (pag) gene or a close homolog was detected in 16 of these samples. For the other three pathogen groups, the frequency of detection was much lower, and none of the samples were positive with both the phylogenetic and virulence gene primer sets. PMID- 16696702 TI - Composition of bacillus species in aerosols from 11 U.S. cities. AB - A PCR-based heteroduplex assay was used to determine the presence and composition of Bacillus species in 11,059 Environmental Protection Agency PM2.5 aerosol samples from 11 U.S. cities. The assay differentiated three groups: Type A containing Bacillus anthracis and very closely related, often pathogenic, Bacillus cereus and Bacillus thuringiensis strains; Type B containing other B. cereus and B. thuringiensis strains; and a third group of more-distantly related Bacillus species. Eight of the 11 cities were positive for Bacillus species in 50% or more of the samples, and the percent of aerosol samples that contained the HD Type A group ranged from 3% to 32%. Cities from the eastern half of the United States generally contained a higher frequency and broader diversity of Bacillus species than the western half of the United States. Positive samples were detected throughout the year. These results have implications for pathogen detection in environmental samples, understanding the natural evolution of new pathogenic strains, and incidence of infection caused by strains of the B. cereus subgroup. PMID- 16696703 TI - Evaluation of variation in control region sequences for Hispanic individuals in the SWGDAM mtDNA data set. AB - The Scientific Working Group on DNA Analysis Methods (SWGDAM) Hispanic data set was analyzed to determine the diversity, phylogeny, and relevant single nucleotide polymorphisms (SNPs) that describe haplogroup patterns for Hispanic Americans (N=686), and to assess the degree of admixture regarding mitochondrial DNA (mtDNA). The largest component of admixture based on mtDNA analysis derives from the four major haplogroups previously observed in Native American ancestry, including A (29.3%), B (15.7%), C (20.6%), and D (4.8%). European (17.8%) and African (11.8%) haplogroups also were observed within this data set. Hispanic SWGDAM samples from the southwest, compared with other SWGDAM Hispanic samples, were observed to have a greater percent of Native American haplogroups present (79.9%), and fewer African American haplogroups (4.5%). A total of 234 SNPs were observed in the data set, including 36 newly reported variable positions. These SWGDAM Hispanic data set SNPs ranged from having 1 to 31 changes (Length=L) on the phylogenetic tree, with site 16519 being the most variable. On average, there were 3.9 character changes for each variable position on the tree. The most variable sites (with 13 or more changes each listed from fastest to slowest) observed were 16519 (L=31), 16189 (L=23), 152 (L=23), 16311 (L=19), 146 (L=17), 195 (L=17), 16093 (L=15), 16362 (L=14), 16129 (L=13), 150 (L=13), and 153 (L=13). These sites are consistent with other reports on highly variable positions. A total of 27 SNPs were chosen to identify all clusters containing 1% (N=7) or more individuals in the SWGDAM Hispanic data set. The descriptive analyses revealed that the SWGDAM Hispanic data set is similar to published Native American and Hispanic data sets. PMID- 16696704 TI - Enhanced elution of sperm from cotton swabs via enzymatic digestion for rape kit analysis. AB - This report describes development of a method for enhanced cell elution from cotton swabs. The method exploits an enzyme mixture for digestion of the cotton to remove intact cells, and can be utilized in conjunction with or to circumvent conventional differential extraction (DE). Samples digested with Aspergillus niger cellulase yielded sperm cell recoveries (18+/-3.5%) similar to conventional DE buffer (23+/-7.8%) while providing intact epithelial cells. Storage time affected the concentration of enzyme required for optimal sperm cell recovery, with longer times requiring increased cellulase concentrations. Cellulase from A. niger yielded a twofold enhancement in sperm cell elution over buffer alone, and preliminary testing of higher activity cellulases from Trichoderma reesei and Trichoderma viride showed even greater enhancement. These results indicate that cellulose-digesting enzymes enhance the release of sperm and epithelial cells from a cotton swab over buffer alone, providing for efficient DNA analysis. PMID- 16696705 TI - Genetic structure among 38 populations from the United States based on 11 U.S. core Y chromosome STRs. AB - A DNA database consisting of the 11 Y chromosome short-tandem-repeat (Y-STR) recommended by the Scientific Working Group on DNA Analysis Methods is constructed for 2517 individuals from 38 populations in the United States. The population samples derive from five ethnic groups currently living in 10 states. A multidimensional scaling (MDS) plot places the populations into four discrete clusters (African Americans (AA), European Americans (EA), Hispanic Americans (HA), and Asian Americans (SA)) and one dispersed cluster of Native Americans. An analysis of molecular variance (AMOVA) indicates that a large proportion of the total genetic variance is partitioned among ethnic groups (24.8%), whereas only a small amount (1.5%) is found among-populations within ethnic groups. Separate AMOVA analyses within each ethnic group show that only the NA sample contains statistically significant among-population variation. Pair wise population differentiation tests do uncover heterogeneity among EA and among HA populations; however, this is due to only a single sample within each group. The analyses support the creation of AA, EA, HA, and Asian American databases in which samples from different geographic regions within the United States are pooled. We recommend that separate databases be constructed for different NA groups. PMID- 16696706 TI - Forensic analysis of bicomponent fibers using infrared chemical imaging. AB - The application of infrared chemical imaging to the analysis of bicomponent fibers was evaluated. Eleven nominally bicomponent fibers were examined either side-on or in cross-section. In six of the 11 samples, infrared chemical imaging was able to spatially resolve two spectroscopically distinct regions when the fibers were examined side-on. As well as yielding characteristic infrared spectra of each component, the technique also provided images that clearly illustrated the side-by-side configuration of these components in the fiber. In one case it was possible to prepare and image a cross-section of the fiber, but in general the preparation of fiber cross-sections proved very difficult. In five of the 11 samples, the infrared spectra could be used to identify the overall chemical composition of the fibers, according to a published classification scheme, but the fiber components could not be spatially resolved. Difficulties that are inherent to conventional "single-point" infrared spectroscopy, such as interference fringing and sloping baselines, particularly when analyzing acrylic type fibers, were also encountered in the infrared chemical image analysis of bicomponent fibers. A number of infrared sampling techniques were investigated to overcome these problems, and recommendations for the best sampling technique are given. Chemical imaging results were compared with those obtained using conventional fiber microscopy techniques. PMID- 16696707 TI - Chemical profiling of heroin recovered from the North Korean merchant vessel Pong Su. AB - Heroin samples, seized from the North Korean merchant vessel Pong Su in Australian waters, were analyzed to determine geographic origin. Duplicate samples were analyzed by the National Measurement Institute's Australian Forensic Drug Laboratory and the United States Drug Enforcement Administration's Special Testing and Research Laboratory. Alkaloid ratios were determined by both liquid chromatography-diode array detection (LC-DAD) and capillary electrophoresis-diode array detection (CE-DAD) techniques. Acid/neutral manufacturing by-products were determined by solvent extraction followed by gas chromatography-mass spectrometry (GC-MS). Solvents, trapped in the heroin particles during manufacture, were detected by both static headspace GC-MS and purge and trap GC-MS. The alkaloid ratios obtained were consistent with heroin of a Southeast Asian (SEA) origin and principal component analysis of the alkaloid results demonstrated the presence of at least four subgroupings within the seizure. The solvent analysis detected diethyl ether and ethyl acetate, solvents typically seen in SEA heroin. However, the acid/neutral analysis revealed compounds not normally seen in heroin of a SEA origin. Furthermore, sterol-like molecules, always detected in the acid/neutral analysis of SEA heroin, were absent from the Pong Su samples. The Pong Su heroin, although similar to SEA heroin, has sufficient differences to classify it as having an unknown origin at the time of this writing. PMID- 16696708 TI - Stable isotope analyses of heroin seized from the merchant vessel Pong Su. AB - A new type of heroin HCl seized in Australia was examined by stable isotope analysis. The final origin/process classification of these samples by chromatographic signature profiles of the impurity/manufacturing by-products was previously determined to be "unknown" by two independent national laboratories. Various drug enforcement authorities speculated that the heroin might be from a new region or new illicit process due to the unusual chromatographic impurity profiles that were present. Samples from 20 different kilogram packages were examined for isotopic content to determine if the samples fit isotopic patterns of known origins or if they were unique to any known origins. Authentic specimens from Southeast Asian (N=59), Southwest Asia (N=37), South America (N=104), and Mexico (N=21) we concomitantly examined for comparison purposes. Both continuous flow elemental analysis-isotope ratio mass spectrometry and gas chromatography isotope ratio mass spectrometry techniques were utilized. Heroin samples were also converted to morphine, without apparent isotopic fractionation, utilizing methanolic HCl for gas chromatography-isotope ratio mass spectrometry. The Pong Su samples were found to be isotopically and isotopically/alkaloidally distinct from the known origin/process classifications of Southwest Asian, Southeast Asian, South American, and Mexican. PMID- 16696709 TI - Sequential monitoring of burials containing large pig cadavers using ground penetrating radar. AB - Ground-penetrating radar (GPR) was used to monitor 12 pig burials in Florida, each of which contained a large pig cadaver. Six of the cadavers were buried in sand at a depth of 0.50-0.60 m, and the other six were buried at a depth of 1.00 1.10 m and were in contact with the upper surface of a clay horizon. Control excavations with no pig internment were also constructed as blank graves and monitored with GPR. The burials were monitored with GPR for durations of either 12-13 or 21-21.5 months when they were then excavated to correlate the decomposition state of the cadaver with the GPR imagery. Overall, cadavers in sand were easily detected for the duration of this study at 21.5 months, even when completely skeletonized. Conversely, in clay it became increasingly difficult to image the pig cadavers over the first year of burial, even when they still retained extensive soft tissue structures. PMID- 16696710 TI - Cracking the code: a decode strategy for the international business machines punch cards of Korean war soldiers. AB - During the Korean War, International Business Machines (IBM) punch cards were created for every individual involved in military combat. Each card contained all pertinent personal information about the individual and was utilized to keep track of all soldiers involved. However, at present, all of the information known about these punch cards reveals only their format and their significance; there is little to no information on how these cards were created or how to interpret the information contained without the aid of the computer system used during the war. Today, it is believed there is no one available to explain this computerized system, nor do the original computers exist. This decode strategy is the result of an attempt to decipher the information on these cards through the use of all available medical and dental records for each individual examined. By cross referencing the relevant personal information with the known format of the cards, a basic guess-and-check method was utilized. After examining hundreds of IBM punch cards, however, it has become clear that the punch card method of recording information was not infallible. In some cases, there are gaps of information on cards where there are data recorded on personal records; in others, information is punched incorrectly onto the cards, perhaps as the result of a transcription error. Taken all together, it is clear that the information contained on each individual's card should be taken solely as another form of personal documentation. PMID- 16696711 TI - A technique to capture, analyze, and quantify anterior teeth rotations for application in court cases involving tooth marks. AB - Anterior teeth within the human dentition have a specific numerical rotation value. Bite marks show an array of angled indentations, abrasions, microlacerations, and contusions. These marks generally represent the incisal surfaces of the suspect's dentition reflecting the rotation values of the teeth in the dental arch. This study described a method for capturing and analyzing anterior dental rotations. The rotations of individual anterior teeth within the study population were categorized as common, uncommon, and very uncommon according to Allen's classification. In the absence of a large number of incisal patterns present in a bite mark, a single but heavily weighted tooth rotation could be of equal discriminatory potential to several common rotation values. No prevalence studies quantifying individual tooth rotations are available. The measurement of each individual tooth rotation together with its individual discrimination potential will enhance the evaluation of the concordant features observed in bite marks. PMID- 16696712 TI - Use of diagonal teeth measurements in predicting gender in a Turkish population. AB - Mesiodistal and buccolingual measurements, commonly used as a means of sex determination from teeth, can sometimes cause difficulties. The aim of this study is to test whether diagonal measurements can make it possible to take more accurate measurements. The results of diagonal measurements of dental casts taken from 30 males and 30 females have been evaluated by discriminant function statistics. Intra- and interobserver error tests did not indicate any statistically significant differences between the findings of two observers. Seven of the 14 measurements on the maxilla and 10 of the 14 measurements on the mandible have been found to be significantly greater in males. According to the results of the stepwise discriminant function statistics, the most contributory measurements to the function were upper first incisor mesiobuccal-distolingual (MBDL) and distobuccal-mesiolingual, lower second incisor MBDL, and lower canine MBDL. The highest reliability was obtained in MBDL measurements. It was realized that diagonal measurements of teeth, especially of canines, revealed clear dimorphic differences. Classification accuracy was found to be 83.3% for total sample, 78.3 for upper jaw, and 85.0% for the lower jaw. Accuracy rate was higher in the lower teeth. Commonly seen orthodontic anomalies, such as tooth rotations, crowding, attritions, deep dentin-enamel junction defects, and certain types of fillings, could make it difficult to correctly take width measurements or could cause other mistakes to occur. This explains the reason why this research has been considered to be of some use in diagonal measurements, which is an accurate method, particularly when employed for the front teeth. PMID- 16696713 TI - Detection and classification of composite resins in incinerated teeth for forensic purposes. AB - The great demand for esthetic restorations has resulted in placement of large numbers of composite resin fillings. The popularity of these materials is reflected in the quantity and variety of resin brands currently on the market. The ability to distinguish resin brands can aid in positive identification of burn victims, assuming that appropriate dental records exist. Scanning electron microscopy/energy dispersive X-ray spectroscopy (SEM/EDS) was used to characterize the structure and composition of 10 modern resins. The structure of each resin was unique to manufacturer, and elemental analysis allowed separation into distinct groups. These 10 resins were also placed in extracted teeth and incinerated at 900 degrees C for 30 min, simulating near cremation conditions. The resins were identifiable by SEM/EDS after incineration, and the elemental composition remained almost unchanged. The data produced are immediately useful for resin identification in forensics, and comparative analysis can be readily performed using standard equipment. This work represents the initial stage of database generation. PMID- 16696714 TI - The development and application of a diatom-based quantitative reconstruction technique in forensic science. AB - Diatoms are a group of unicellular algae that have been recorded and classified for over 200 years and have been used in a range of applications in forensic science. We have developed a quantitative diatom-based reconstruction technique to confirm drowning as a cause of death and localize the site of drowning in two recent, high-profile, case studies. In both case studies we collected diatom samples from the local and/or regional area to act as a control in the examination of diatom assemblages associated with lungs and clothing. In Case Study 1 the modern analog technique suggested that all lung and clothing samples have statistically significant similarities to control samples from shallow water habitats. In Case Study 2, the analog matching suggested that the majority of lung samples show a statistically significant relationship to samples from a pond, indicating that this was the drowning medium. PMID- 16696715 TI - Amended cause and manner of death certification: a six-year review of the New Mexico experience. AB - Little is known about the amendment of death certificates (DCs) issued by medical examiners and coroners. This retrospective study examined why, how, and with what frequency cause and manner of death were amended on DCs issued by forensic pathologists over a 6-year period at the New Mexico Office of the Medical Investigator. Approximately 1% of DCs had either cause or manner of death amendments, with arteriosclerotic cardiovascular disease and intoxicants the most commonly amended and resulting causes of death, respectively. There was a significant association between manner of death and number of DCs amended (p<0.001). By percent, natural and suicide DCs were the most frequently amended. The way in which manner of death changed was significantly associated with the amount of time elapsed between DCs (p=0.04). Toxicology was the most common reason for DC amendment. PMID- 16696716 TI - Influence of substrate tissue type on larval growth in Calliphora augur and Lucilia cuprina (Diptera: Calliphoridae). AB - The size of fly larvae is an important variable in the use of these insects to estimate postmortem interval. Furthermore, the nutritional intake of larvae is likely to vary subject to the part of a corpse on which they are feeding. A study was therefore conducted to investigate the effect of type of food substrate on larval growth in two species of forensically important Australian blowflies. After collection on sheep's liver in the laboratory, different groups of larvae of Lucilia cuprina (Wiedemann) and Calliphora augur (Fabricius) were grown on sheep's liver, meat, and brains, and their body lengths compared. Results indicated that the development of larvae fed sheep's liver was adversely affected compared with larvae fed meat and brain; they moulted later, reached maximum length more slowly and sometimes produced significantly smaller pupae. These findings, similar to those of another recent study, have obvious implications for postmortem interval determinations. Estimates may be considerably skewed if the site of collection of larvae at a death scene contains tissue types different to those used in reference experiments. We therefore recommend caution in forensic analyses that interpret crime scene data using developmental studies performed with a single type of larval food substrate. PMID- 16696717 TI - Fatal capnocytophaga infection associated with splenectomy. AB - A case of fatal sepsis due to Capnocytophaga species is described. Capnatophaga canimorsus and C. cynodegmi can cause localized wound infections and/or systemic infections in people who have been bitten, licked, scratched, or merely exposed to cats or dogs, especially splenectomized individuals. A thorough social, medical, and surgical history, the clinical presentation, and cultures are important in making the diagnosis of Capnocytophaga infections. It is important that the forensic pathologist be aware of this zoonotic disease. PMID- 16696718 TI - Sudden death from tubercular myocarditis. AB - Tuberculous myocarditis is a rare finding. We present the case of a 33-year-old woman who was in good health and who died suddenly at home. Autopsy and histopathologic examinations revealed granulamatous lesions in the myocardium, lungs, lymph nodes, liver, and spleen. No fast acid bacilli were demonstrated on histological examination. The presence of a Mycobacterium tuberculosis DNA complex was identified using a polymerase chain reaction (PCR) on formalin-fixed paraffin-embedded histological samples. An HIV test carried out on the blood obtained during the autopsy was negative according to the DNA amplification technique (PCR) and enzyme-linked immunosorbent assay serological test. We hypothesize that the mechanism of death was severe ventricular arrhythmia due to granulomatous proliferation in the structures of the interventricular septum. PMID- 16696719 TI - Malnutrition, a rare form of child abuse: diagnostic criteria. AB - Infantile malnutrition is often difficult to diagnose as it is rarely observed in industrialized countries. It may be associated with physical violence or occur in isolation. The essential clinical sign is height and weight retardation, but malnutrition also causes a variety of internal and bone lesions, which lead to neuropsychological sequelae and death. We report a rare case of death by malnutrition in a female child aged 6 1/2 months. The infant presented height and weight growth retardation and internal lesions related to prolonged protein energy malnutrition (fat and muscle wasting, thymic atrophy, liver steatosis) resulting in a picture of marasmus or kwashiorkor. We detail the positive and negative criteria that established the diagnosis of abuse, whereas the parents had claimed a simple dietary error. PMID- 16696720 TI - Alligator attacks in southwest Florida. AB - The American alligator inhabits bodies of fresh water in Florida and other southeastern states. Although attacks on pets are frequent, alligator attacks on humans are relatively rare because of the animal's natural fear of man. Because of the rarity of attacks on humans, the pathologic findings and pathophysiology of death in such cases have not been well characterized in the literature. We report three cases of fatal alligator attacks that occurred in southwest Florida, each with different pathologic findings and mechanisms of death. Although the cause of death in each case was attributed to the alligator attack, the mechanisms of death differed and included exsanguination because of amputation of an extremity, overwhelming sepsis, and drowning. These cases illustrate the varied pathophysiologies associated with deaths due to alligator attacks on humans and the features that distinguish alligator bites from those of other aquatic predators. PMID- 16696721 TI - The presence of low IQ and mental retardation among murder defendants referred for pretrial evaluation. AB - After an overview of definitions of mental retardation and recent case law regarding mental retardation and the death penalty, this paper presents a study of factors associated with a mental retardation (MR) diagnosis among murder defendants. Subjects with a full-scale IQ< or =70 (n=42) were compared with other pretrial murder defendants (n=228) referred for forensic evaluation over a 5-year period. Subjects with an IQ< or =70 who were diagnosed with MR were compared with subjects with an IQ< or =70 who did not receive this diagnosis. Female murder defendants were more likely to receive a diagnosis of MR (p=0.03). MR was also more commonly diagnosed in subjects with an Axis I cognitive disorder (p=0.018). Having an IQ< or =70 was more common in subjects with a psychotic and substance use disorder (p=0.03) and did not necessarily lead to a diagnosis of MR in this subgroup. Implications for diagnosing MR among murder defendants are discussed. PMID- 16696722 TI - The paraphilia-related disorders: an investigation of the relevance of the concept in sexual murderers. AB - Paraphilic disorders (PAs) and sexual preoccupation are known risk factors for recidivism in sexual offenders. Nonparaphilic sexual excessive behaviors-so called paraphilia-related disorders (PRDs), like paraphilias, are also characterized by sexual preoccupation and volitional impairment and can be diagnosed in paraphilic men. The prevalence and clinical significance of PRDs in sexual homicide perpetrators, however, is unknown. We investigated the relationship between PAs and PRDs retrospectively in a sample of 161 sexual murderers. Four groups were compared: men without a PA or a PRD diagnosis, men with at least one PRD but no PA, men with at least one PA but no PRD, and finally, those with a combination of both (PA+PRD). The PA+PRD group had the most lifetime cumulative sexual impulsivity disorders, more developmental problems, the highest persistent frequency of sexual activity, the highest number of previous sexual offences, more sexual sadism, and compulsive masturbation. Men of the PRD subsample had suffered more from childhood sexual abuse, showed more promiscuity, psychopathy, and alcohol problems. The use of the PRD concept in this special offender group should be further investigated with prospectively designed studies. PMID- 16696723 TI - An evaluation of matching unknown writing inks with the United States International Ink Library. AB - Utilizing a database of standards for forensic casework is a valuable resource. Undoubtedly, as more standards (and corresponding information about the specimens) are collected, there is a greater certainty of identification when a questioned and a known item cannot be distinguished after a series of analyses. The United States Secret Service and the Internal Revenue Service National Forensic Laboratory jointly maintain the largest known forensic collection of writing inks in the world, which is comprised of over 8500 ink standards collected worldwide, dating back to the 1920s. This study was conducted to evaluate the reliability of matching arbitrarily purchased pens with known inks from a database. One hundred pens were randomly obtained from a variety of sources and their respective ink compositions were compared with standards. Eighty-five of the inks were determined to be suitable for comparison utilizing optical examinations and thin-layer chromatography. Three of the inks did not match any of the specimens on record; one of these inks was similar to an ink from an identical brand of pen that was in the database, but had a modified formulation. PMID- 16696724 TI - Commentary On: Kovarik CL, Stewart D, Cockerell CJ, Barnard J. Forensic dermatopathology and internal disease. J Forensic Sci 2005;50(1):154-8. PMID- 16696725 TI - Letter to the editor--nomenclature and allele repeat structure update for the Y STR locus GATA H4. PMID- 16696726 TI - A new allele of the short tandem repeat locus D21S11 in a Venezuelan population. PMID- 16696727 TI - Allele frequencies for three STR loci D10S1418, D8S1477, and D10S1426 in a Chinese population. PMID- 16696728 TI - Population genetics for five STR loci in Tibetan group of Chinese population. PMID- 16696729 TI - Population data on HLA-DQA1, LDLR, GYPA, HBGG, D7S8, and GC PCR-based loci in Serbia. PMID- 16696730 TI - Population data for four population groups from the United States for the eleven Y-chromosome STR loci recommended by SWGDAM. PMID- 16696731 TI - Genetic polymorphisms at four STR loci from the HLA region in a Venezuelan population. AB - POPULATIONS: Whole blood samples from 74 unrelated healthy individuals were collected. The donors' sample included Venezuelan mestizos from various regions of the country, but mostly from the resident population of Caracas City. A Venezuelan mestizo is the offspring of a mating between a native Venezuelan and a person born in Europe, mainly in Spain. PMID- 16696732 TI - Population genetics of two STR loci D2S1346 and D2S1353 in a Han population of Chinese. PMID- 16696733 TI - Allele frequency profile of three STR loci in nine North Indian populations. AB - POPULATION: Bhargavas (n=120), Chaturvedis (n=120), Brahmins (n=120), Muslim Sunni (n=120), Muslim Shiya (n=120), Kayastha (n=120), Mathurs (n=120), Rastogies (n=120), and Vaish (n=120). PMID- 16696734 TI - Genetic analysis of six STR polymorphisms in a southern Chinese population. AB - POPULATION: A total of 105 unrelated and healthy individuals from the Han ethnic group of Nanning city in south China. PMID- 16696735 TI - Genetic polymorphism for the PowerPlex 16 system from the Chinese Tujia Ethnic Minority Group. AB - POPULATIONS: The population studied was the Tujia population, living in the Hunan Province, China. Their ancestors had lived in the region for at least three generations. As a Chinese minority group, the Tujia ethnic group, with a population of 8,028,133 (year of 2000), is mainly distributed in the Wuling mountain area in the three provinces of Hunan, Hubei, and Sichuan. The Tujia population have their own language, which belongs to the Tibetan-Myanmese Language Group of the Chinese-Tibetan Language Family, but the large majority have come to speak the Han and Miao languages, now that the Tujias have been largely assimilated. PMID- 16696737 TI - Performance-based measures of physical function for high-function populations. AB - OBJECTIVES: To improve and broaden the applicability of performance-based measures of function for use in clinical and research settings. DESIGN: Cross sectional repeated-measures study. SETTING: Research clinic. PARTICIPANTS: Population-based sample of Japanese Americans without significant functional impairments aged 35 to 55 and 56 to 71 (N=203). MEASUREMENTS: Performance-based measures of physical function, including range of standard tests, newer automated measures of balance and strength and data on cognitive function, lifestyle, medical history, and physical activity. RESULTS: Of the nonplatform balance measures, only the one-leg stand was reliable (reliability coefficient (rc) =0.69) and able to discriminate between functional levels. Combining the Fourth National Health and Nutrition Examination Survey protocol of folded arm position while standing on a foam pad with the more-sophisticated balance platform test had the highest reliability and discrimination. With the strength chair, high rcs (0.88-0.96) were found for upper and lower extremity tests. Timed chair stands, a test of lower extremity and central strength were reliable. The 6-minute walk had a high rc (0.90). CONCLUSION: Many performance tests used today are not reliable. Only a few discriminate between the most highly functioning individuals and individuals with good function. Thus, a new recommended battery includes unassisted single-leg stand, balance platform "foam pad, eyes closed," elbow flexion and knee extension strength (strength chair), grip strength, timed chair stands, and the 6-minute walk. These simple performance-based tests have good reliability and discrimination across the range of function and can be used in most clinical and research settings to quickly assess global functional level. PMID- 16696738 TI - Meaningful change and responsiveness in common physical performance measures in older adults. AB - OBJECTIVES: To estimate the magnitude of small meaningful and substantial individual change in physical performance measures and evaluate their responsiveness. DESIGN: Secondary data analyses using distribution- and anchor based methods to determine meaningful change. SETTING: Secondary analysis of data from an observational study and clinical trials of community-dwelling older people and subacute stroke survivors. PARTICIPANTS: Older adults with mobility disabilities in a strength training trial (n=100), subacute stroke survivors in an intervention trial (n=100), and a prospective cohort of community-dwelling older people (n=492). MEASUREMENTS: Gait speed, Short Physical Performance Battery (SPPB), 6-minute-walk distance (6MWD), and self-reported mobility. RESULTS: Most small meaningful change estimates ranged from 0.04 to 0.06 m/s for gait speed, 0.27 to 0.55 points for SPPB, and 19 to 22 m for 6MWD. Most substantial change estimates ranged from 0.08 to 0.14 m/s for gait speed, 0.99 to 1.34 points for SPPB, and 47 to 49 m for 6MWD. Based on responsiveness indices, per-group sample sizes for clinical trials ranged from 13 to 42 for substantial change and 71 to 161 for small meaningful change. CONCLUSION: Best initial estimates of small meaningful change are near 0.05 m/s for gait speed, 0.5 points for SPPB, and 20 m for 6MWD and of substantial change are near 0.10 m/s for gait speed, 1.0 point for SPPB, and 50 m for 6MWD. For clinical use, substantial change in these measures and small change in gait speed and 6MWD, but not SPPB, are detectable. For research use, these measures yield feasible sample sizes for detecting meaningful change. PMID- 16696739 TI - Chronic kidney disease and functional limitation in older people: health, aging and body composition study. AB - OBJECTIVES: To assess whether chronic kidney disease (CKD) is independently associated with incident physical-function limitation. DESIGN: Prospective cohort study. SETTING: Two sites: Pittsburgh, Pennsylvania, and Memphis, Tennessee. PARTICIPANTS: Two thousand one hundred thirty-five men and women aged 70 to 79 without functional limitation at baseline from the Health, Aging and Body Composition Study. MEASUREMENTS: Functional limitation was defined as difficulty in walking one-quarter of a mile or climbing 10 steps on two consecutive reports 6 months apart (in the same function). Kidney function was measured using serum cystatin C. Estimated glomerular filtration rate (eGFR), using the Modification of Diet in Renal Disease formula (<60 versus > or =60 mL/min per 1.73 m(2)), was a secondary predictor. Muscle strength, lean body mass according to dual energy x ray absorptiometry, comorbidity, medication use, and inflammatory markers were evaluated as covariates. RESULTS: Persons in the highest (> or =1.13 mg/L) quartile of cystatin C experienced a significantly higher risk of developing functional limitation than those in the lowest (<0.86 mg/L) quartile (hazard ratio (HR)=1.70, 95% confidence interval (CI)=1.40-2.07). The association between the fourth cystatin C quartile and functional limitation remained after adjustment for demographics, lean body mass, comorbidity, muscle strength, and gait speed (HR=1.41, 95% CI=1.13-1.75), although the association was attenuated after adjustment for markers of inflammation (HR=1.15, 95% CI=0.90-1.46). Similar results were found for eGFR less than 60 mL/min per 1.73 m(2), although the association with functional limitation remained after adjustment for inflammatory markers (HR=1.30, 95% CI=1.08-1.56). CONCLUSION: CKD is associated with the development of functional impairment independent of comorbidity, body composition, and tests of strength and physical performance. The mechanism may be related to a heightened inflammatory state in CKD. PMID- 16696740 TI - Alcohol use, comorbidity, and mortality. AB - OBJECTIVES: To examine the combined influence of alcohol use and comorbidity on 20-year mortality in older adults (average age 66 at the time of the baseline survey). DESIGN: Longitudinal analysis of a national probability sample-based cohort study. SETTING: Data sources were the National Health and Nutrition Examination Survey I (NHANES I), 1971-1974, and the NHANES Epidemiologic Followup Survey, 1992. PARTICIPANTS: Four thousand six hundred ninety-one adults aged 60 and older who provided data on alcohol use. MEASUREMENTS: The prevalence of at risk drinking in older adults in the United States and the 20-year all-cause mortality risk associated with it. At-risk drinking status was determined from amount of alcohol consumed and comorbidities, using a previously validated method. RESULTS: The prevalence of at-risk drinking in the United States between 1971 and 1974 in older adults was 10% (18% of men, 5% of women). The majority of at-risk drinkers were identified as such because of their use of alcohol in amounts deemed risky in the presence of relevant comorbidities (69%) (e.g., drinking 2-3 drinks per day and having gout or anxiety or taking a medication for pain). In men, at-risk drinking was associated with higher mortality rates than not-at-risk drinking (hazard ratio=1.20, 95% confidence interval=1.01-1.41); abstinence was not associated with greater mortality. In women, neither abstinence nor at-risk drinking was associated with greater mortality rates. CONCLUSION: In this first, large population-based study of older adults examining the mortality risks of alcohol use and comorbidity, at-risk drinking was associated with greater mortality rates in men. These findings suggest that a lower threshold of alcohol use should be recommended for older adults with specific comorbidities to reduce mortality risks. PMID- 16696741 TI - Ankle brachial index as a predictor of cognitive impairment in the general population: ten-year follow-up of the Edinburgh Artery Study. AB - OBJECTIVES: To determine whether the ankle brachial index (ABI, a marker of generalized atherosclerosis) is associated with cognitive impairment after 10 years in older people. DESIGN: Cohort study (Edinburgh Artery Study). SETTING: Eleven general practices in Edinburgh, Scotland. PARTICIPANTS: Seven hundred seventeen men and women aged 55 to 74 from the general population, followed for 10 years. MEASUREMENTS: ABI measured at baseline and major cognitive functions (including premorbid function using the National Adult Reading Test, NART) tested after 10 years. RESULTS: After adjustment for age and sex, a low ABI was associated with lower scoring (bottom tertile vs top tertile) on Raven's Matrices (odds ratio (OR)=1.6, 95% confidence interval (CI) =1.0-2.6), Verbal Fluency (OR =1.8, 95% CI =1.1-3.0), and Digit Symbol Test (OR =2.3, 95% CI =1.3-4.2), suggesting that the ABI is predictive of poorer performance in nonverbal reasoning, verbal fluency, and information processing speed. The association between ABI and the Digit Symbol Test remained significant after further adjustment for premorbid cognitive function (tested using the NART), suggesting that the ABI is also predictive of decline in information processing speed (from premorbid ability to that measured here in older age). CONCLUSION: The ABI may be useful in identifying older individuals at higher risk of cognitive impairment. In the future, preventive measures developed to target individuals with a low ABI should consider measures to reduce vascular-related cognitive decline as well as cardiovascular events, in an effort to reduce the incidence and consequences of subsequent cognitive impairment and dementia. PMID- 16696742 TI - Limited literacy in older people and disparities in health and healthcare access. AB - OBJECTIVES: To determine the relationship between health literacy, demographics, and access to health care. DESIGN: Cross-sectional study, Health, Aging and Body Composition data (1999/2000). SETTING: Memphis, Tennessee, and Pittsburgh, Pennsylvania. PARTICIPANTS: Two thousand five hundred twelve black and white community-dwelling older people who were well functioning at baseline (without functional difficulties or dementia). MEASUREMENTS: Participants' health literacy was assessed using the Rapid Estimate of Adult Literacy in Medicine. Scores were categorized into 0 to sixth-, seventh- to eighth-, and ninth-grade and higher reading levels (limited health literacy defined as <9th grade). Participants' demographics, socioeconomic status, comorbidities, and three indicators of healthcare access (whether they had a doctor/regular place of medical care, an influenza vaccination within the year, or insurance for medications) were also assessed. RESULTS: Participants' mean age was 75.6, 52% were female, 38% were black, and 24% had limited health literacy. After adjusting for sociodemographics, associations remained between limited health literacy and being male, being black, and having low income and education, diabetes mellitus, depressive symptoms, and fair/poor self-rated health (P<.02). After adjusting for sociodemographics, health status, and comorbidities, older people with a sixth grade reading level or lower were twice as likely to have any of the three indicators of poor healthcare access (odds ratio=1.96, 95% confidence interval=1.34-2.88). CONCLUSION: Limited health literacy was prevalent and was associated with low socioeconomic status, comorbidities, and poor access to health care, suggesting that it may be an independent risk factor for health disparities in older people. PMID- 16696743 TI - Continuous positive airway pressure reduces subjective daytime sleepiness in patients with mild to moderate Alzheimer's disease with sleep disordered breathing. AB - OBJECTIVES: Studies have reported that 33% to 70% of patients with Alzheimer's disease (AD) have sleep-disordered breathing (SDB). Continuous positive airway pressure (CPAP) treatment has been shown to reduce daytime sleepiness and improve health-related quality of life in nondemented older people with SDB. The effect of therapeutic CPAP treatment on daytime sleepiness in patients with mild moderate AD with SDB was assessed. DESIGN: Randomized, double-blind, placebo controlled trial. SETTING: Patients' home and the University of California San Diego, General Clinical Research Center, J. Christian Gillin Laboratory of Sleep and Chronobiology. PARTICIPANTS: Thirty-nine community-dwelling elderly patients with mild-moderate probable AD with SDB. INTERVENTION: Patients were randomly assigned to receive 6 weeks of therapeutic CPAP or 3 weeks of sham CPAP followed by 3 weeks of therapeutic CPAP. MEASUREMENTS: Epworth Sleepiness Scale (ESS) was administered at baseline, 3 weeks, and 6 weeks. Changes in daytime sleepiness in subjects who received optimal therapeutic CPAP were compared with changes in the sham CPAP group. RESULTS: Within the therapeutic CPAP group, ESS scores were reduced from 8.89 during baseline to 6.56 after 3 weeks of treatment (P=.04) and to 5.53 after 6 weeks of treatment (P=.004). In the sham CPAP group, there was no significant difference after 3 weeks of sham CPAP but a significant decrease from 7.68 to 6.47 (P=.01) after 3 weeks of therapeutic CPAP. CONCLUSION: These data provide evidence of the effectiveness of CPAP in reducing subjective daytime sleepiness in patients with AD with SDB. PMID- 16696744 TI - Safety and efficacy of teriparatide in elderly women with established osteoporosis: bone anabolic therapy from a geriatric perspective. AB - OBJECTIVES: To assess the safety and efficacy of teriparatide in patients aged 75 and older and compare these findings with those of women younger than 75 using data from the Fracture Prevention Trial (FPT). DESIGN: The FPT was a randomized, multicenter, double-blind, placebo-controlled study. SETTING: The FPT multicenter international study. PARTICIPANTS: Postmenopausal women aged 42 to 86 were randomized to placebo (N=544) or teriparatide 20 mug (N=541) by daily self injection for a median of 19 months. Patients received daily oral supplements of 1,000 mg calcium and 400 to 1,200 IU vitamin D. For this analysis, subgroups were defined according to patient age younger than 75 (N=841) and 75 and older (N=244). MEASUREMENTS: The effects of teriparatide on bone mineral density (BMD) of the lumbar spine and femoral neck; the incidence of new vertebral and new nonvertebral fragility fractures; bone turnover markers, including bone-specific alkaline phosphatase; and urinary deoxypyridinoline corrected for creatinine clearance, as well as the safety of teriparatide, were investigated. RESULTS: There were no significant treatment-by-age interactions for the bone turnover markers, femoral neck BMD, vertebral fractures, nonvertebral fragility fractures, height loss, hyperuricemia, or hypercalcemia. A significant treatment-by-age interaction for lumbar spine BMD (P=.08) was due to an increase in BMD observed in the placebo group aged 75 and older. There were no treatment-by-age interactions for important treatment-emergent adverse events (TEAEs), including back pain, nausea, leg cramps, and dizziness. The most important TEAEs in women aged 80 and older (23 patients from the placebo group and 25 patients from the teriparatide group) were also reviewed; no unexpected TEAEs were found in the patients treated with teriparatide. These results indicate that the clinical effects of teriparatide were consistent in the older and younger women. CONCLUSION: Age does not affect the safety and efficacy of teriparatide in postmenopausal women with osteoporosis. PMID- 16696745 TI - Between-meal risedronate does not alter bone turnover in nursing home residents. AB - OBJECTIVES: To assess the effect of between-meal weekly risedronate and daily calcium 630 mg and vitamin D 400 IU on bone turnover markers. DESIGN: Randomized,double-blind,placebo-controlled trial. SETTING: Skilled nursing home (NH). PARTICIPANTS: Sixty skilled-NH residents (46 men, 14 women), mean age+/ standard deviation of 76+/-6, were randomized to receive risedronate 30 mg (n=31) or matching placebo (n=29) once weekly for 12 weeks. All received 315 mg calcium with 200 IU vitamin D twice daily. MEASUREMENTS: Bone-specific alkaline phosphatase (BSAP), N-telopeptide of type 1 collagen (NTx), 25-hydroxyvitamin D (25OHD), and parathyroid hormone were measured at baseline and 6 and 12 weeks. RESULTS: Risedronate reduced BSAP significantly more than placebo (P<.05) at 6 weeks but not at 12 weeks; no treatment effect on serum NTx was observed. Defining hypovitaminosis D as a serum 25OHD concentration below 32 ng/mL, 50 of 53 (94%) study participants were low at baseline (mean 25OHD 19 ng/mL). Vitamin D levels remained insufficient in 74% of participants after 12 weeks. CONCLUSION: In this NH population, weekly risedronate administered using a between-meal dosing schedule reduced serum BSAP at 6 weeks of treatment; this effect was not observed at 12 weeks. The overall lack of change in bone turnover markers suggests that this risedronate dose and schedule would not be expected to increase bone density or reduce fracture risk in this population. Hypovitaminosis D was common and not reliably corrected by 400 IU of vitamin D daily. Despite an extremely high osteoporotic fracture risk in NH residents, additional study is required to determine under which conditions pharmacological treatment is efficacious in this population and define approaches that assure vitamin D repletion. PMID- 16696746 TI - Caregiver characteristics are associated with neuropsychiatric symptoms of dementia. AB - OBJECTIVES: To determine whether caregiver characteristics are independently associated with neuropsychiatric symptoms of dementia (NPS) after accounting for patient characteristics. DESIGN: Cross-sectional analysis of data from the Medicare Alzheimer's Disease Demonstration and Evaluation study. SETTING: Community-dwelling residents in eight U.S. cities. PARTICIPANTS: Five thousand seven hundred eighty-eight patients with dementia and their caregivers. MEASUREMENTS: Caregivers were asked about the presence of 12 NPS in patients with dementia. Caregiver predictors included age, sex, education, income, marital status, relationship to the patient, whether they lived with patient, number of hours per week spent caregiving, self-reported health, dependency in activities of daily living (ADLs) and instrumental activities of daily living (IADLs), depression, and burden. Multivariate linear regression was used to determine which caregiver characteristics were independently associated with reports of more NPS in patients after controlling for the patient's age, sex, dementia severity, level of ADL dependency, and dementia type. RESULTS: Caregivers were on average 64 years old, 72% female, and 49% were the spouse of the patient (32% wives, 17% husbands). The mean burden score of caregivers was 15 (range 0-32, with higher scores indicating more burden), and 32% had significant depressive symptoms. Patients were on average 79 years old, 60% were female, and most had moderate to severe dementia. The mean number of NPS+/-standard deviation was 4.8+/-2.8. After adjusting for patient characteristics, caregivers who were younger, less educated, more depressed, more burdened, or spent more hours per week giving care reported more NPS in care recipients (all P< or =.005). CONCLUSION: Certain caregiver characteristics are associated with NPS, independent of patient characteristics, including dementia severity. Clinicians should consider the dynamics between patients and caregivers when managing NPS. Understanding how different caregiver characteristics influence NPS may help tailor caregiver education and interventions. PMID- 16696747 TI - Systemic inflammatory response syndrome in nosocomial bloodstream infections with Pseudomonas aeruginosa and Enterococcus Species: comparison of elderly and nonelderly patients. AB - OBJECTIVES: To determine whether the systemic inflammatory response syndrome (SIRS), clinical course, and outcome of monomicrobial nosocomial bloodstream infection (BSI) due to Pseudomonas aeruginosa or Enterococcus spp. is different in elderly patients than in younger patients. DESIGN: Historical cohort study. SETTING: An 820-bed tertiary care facility. PARTICIPANTS: One hundred twenty seven adults with P. aeruginosa or enterococcal BSI. MEASUREMENTS: SIRS scores were determined 2 days before the first positive blood culture through 14 days afterwards. Elderly patients (> or =65, n=37) were compared with nonelderly patients (<65, n=90). Variables significant for predicting mortality in univariate analysis were entered into a logistic regression model. RESULTS: No difference in SIRS was detected between the two groups. No significant difference was noted in the incidence of organ failure, 7-day mortality, or overall mortality between the two groups. Univariate analysis revealed that Acute Physiology And Chronic Health Evaluation (APACHE) II score of 15 or greater at BSI onset; adjusted APACHE II score (points for age excluded) of 15 or greater at BSI onset; and respiratory, cardiovascular, renal, hematological, and hepatic failure were predictors of mortality. Age, sex, use of empirical antimicrobial therapy, and infection with imipenem-resistant P. aeruginosa or vancomycin resistant enterococci did not predict mortality. Multivariate analysis revealed that hematological failure (odds ratio (OR)=8.1, 95% confidence interval (CI)=2.78-23.47), cardiovascular failure (OR=4.7, 95% CI=1.69-13.10), and adjusted APACHE II > or = 15 at BSI onset (OR=3.1, 95% CI=1.12-8.81) independently predicted death. CONCLUSION: Elderly patients did not differ from nonelderly patients with respect to severity of illness before or at the time of BSI. Elderly patients with pseudomonal or enterococcal BSIs did not have a greater mortality than nonelderly patients. PMID- 16696748 TI - A randomized trial of a multicomponent home intervention to reduce functional difficulties in older adults. AB - OBJECTIVES: To test the efficacy of a multicomponent intervention to reduce functional difficulties, fear of falling, and home hazards and enhance self efficacy and adaptive coping in older adults with chronic conditions. DESIGN: A prospective, two-group, randomized trial. Participants were randomized to a treatment group or no-treatment group. SETTING: Urban community-living older people. PARTICIPANTS: Three hundred nineteen community-living adults aged 70 and older who reported difficulty with one or more activities of daily living. INTERVENTION: Occupational and physical therapy sessions involving home modifications and training in their use; instruction in strategies of problem solving, energy conservation, safe performance, and fall recovery techniques; and balance and muscle strength training. MEASUREMENTS: Outcome measures included self-rated functional difficulties with ambulation, instrumental activities of daily living, activities of daily living, fear of falling, confidence performing daily tasks, and use of adaptive strategies. Observations of home hazards were also conducted. RESULTS: At 6 months, intervention participants had less difficulty than controls with instrumental activities of daily living (P=.04, 95% confidence interval (CI)=-0.28-0.00) and activities of daily living (P=.03, 95% CI=-0.24 to -0.01), with largest reductions in bathing (P=.02, 95% CI=-0.52 to 0.06) and toileting (P=.049, 95% CI=-0.35-0.00). They also had greater self efficacy (P=.03, 95% CI=0.02-0.27), less fear of falling (P=.001, 95% CI=0.26 0.96), fewer home hazards (P=.05, 95% CI=-3.06-0.00), and greater use of adaptive strategies (P=.009, 95% CI=0.03-0.22). Benefits were sustained at 12 months for most outcomes. CONCLUSION: A multicomponent intervention targeting modifiable environmental and behavioral factors results in life quality improvements in community-dwelling older people who had functional difficulties, with most benefits retained over a year. PMID- 16696749 TI - Risk factors and prediction of postoperative delirium in elderly hip-surgery patients: implementation and validation of a medical risk factor model. AB - OBJECTIVES: To evaluate risk factors for postoperative delirium in a cohort of elderly hip-surgery patients and to validate a medical risk stratification model. DESIGN: Prospective cohort study. SETTING: Medical school-affiliated general hospital in Alkmaar, the Netherlands. PARTICIPANTS: Six hundred three hip-surgery patients aged 70 and older screened for risk factors for postoperative delirium. MEASUREMENTS: Predefined risk factors for delirium were assessed on admission. One point was assigned for each of four risk factors present, resulting in three groups: low, intermediate, and high risk. Baseline screening and assessment included the Mini-Mental State Examination, the standardized Snellen test for visual impairment, chart review to determine Acute Physiological and Chronic Health Evaluation II score, and blood urea nitrogen to creatinine ratio. The primary outcome was postoperative delirium, as defined using Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, and Confusion Assessment Method criteria. All patients were screened daily for delirium. RESULTS: Incidence of delirium was 3.8% in the low-risk group (P<.001), 11.1% in the intermediate-risk group (P=.27, relative risk (RR)=3.0), and 37.1% in the high risk group (P<.001, RR=9.8). Cognitive impairment at admission had the highest predictive value for postoperative delirium (coefficient of determination=0.15). Contrary to previous findings, age was an independent predictive factor for delirium. Moreover, postoperative delirium was four times as frequent in acute patients as in elective hip-replacement patients. CONCLUSION: The medical risk factor model is valid for elderly hip-surgery patients. Cognitive impairment, age, and type of admission are important risk factors for delirium in this surgical population. PMID- 16696750 TI - Variation in the ciliary neurotrophic factor gene and muscle strength in older Caucasian women. AB - OBJECTIVES: To determine whether genetic variants in the ciliary neurotrophic factor (CNTF) gene are associated with muscle strength in older women. DESIGN: Cross-sectional analysis of baseline data from the Women's Health and Aging Studies I (1992) and II (1994), complementary population-based studies. SETTING: Twelve contiguous ZIP code areas in Baltimore, Maryland. PARTICIPANTS: Three hundred sixty-three Caucasian, community-dwelling women aged 70 to 79. MEASUREMENTS: Participants were genotyped at the CNTF locus for eight single nucleotide polymorphisms (SNPs), including the null allele rs1800169. The dependent variables were grip strength and the frailty syndrome, identified as presence of three or more of five frailty indicators (weakness, slowness, weight loss, low physical activity, exhaustion). In addition to genotypes, independent variables of body mass index (BMI) and osteoarthritis of the hands were included. RESULTS: Using multivariate linear regression, single SNP analysis identified five SNPs significantly associated with grip strength (P<.05), after adjusting for age, BMI, and osteoarthritis. Haplotype analysis was performed, and a single haplotype associated with grip strength was identified (P<.01). The rs1800169 null allele fully explained the association between this haplotype and grip strength under a recessive model, with individuals homozygous for the null allele exhibiting a 3.80-kg lower (95% confidence interval=1.01-6.58) grip strength. No association was seen between the CNTF null allele and frailty. CONCLUSION: Individuals homozygous for the CNTF null allele had significantly lower grip strength but did not exhibit overt frailty. Larger prospective studies are needed to confirm this finding and extend it to additional populations. PMID- 16696751 TI - Prospective study of fever and pneumonia after flexible fiberoptic bronchoscopy in older people. AB - OBJECTIVES: To assess the frequency of fever and pneumonia after fiberoptic bronchoscopy (FOB) in older people and to evaluate increased risk for these two adverse events with increasing age. DESIGN: Prospective study. SETTING: University hospital system. PARTICIPANTS: Three hundred fifty-eight patients, with 165 (46.1%) patients aged 70 and older, undergoing bronchoscopy. MEASUREMENTS: Indications, abnormal bronchoscopic findings, sampling procedures, final diagnosis, and fever and pneumonia after bronchoscopy. RESULTS: With regard to the indication for bronchoscopy and abnormal bronchoscopic findings, there was no statistical difference between elderly patients (> or =70) and younger patients (<70). Procedures such as forceps biopsy, brushing, and curetting were not performed more often in elderly patients, although final diagnosis of lung cancer was significantly more common in the elderly group (P=.002). The overall incidence of fever after FOB was 6.7%, and that of pneumonia after FOB was 5.6%. The patients aged 70 and older had an incidence of fever after FOB of 3.6% and an incidence of pneumonia of 4.2%, which were not higher than those in patients younger than 70. CONCLUSION: Increasing age is not associated with increasing fever and pneumonia after FOB, and chronological age should not be considered a limiting factor in the decision of whether to perform FOB when it is clinically indicated. PMID- 16696752 TI - Could self-neglect in older adults be a geriatric syndrome? AB - Self-neglect in older adults is a complex phenomenon characterized by inattention to health and hygiene, typically stemming from an inability or unwillingness to access potentially remediating services. Some aspects of self-neglect clinically resemble geriatric syndromes (e.g., falling, incontinence). The literature on self-neglect was comprehensively reviewed and its quality evaluated in the context of considering its candidacy for a geriatric syndrome. MEDLINE (1966 2004) was searched using self-neglect as a keyword. Using a "snowball" sampling strategy, associated terms (e.g., Diogenes' syndrome) were combined, selecting relevant papers and frequently cited references, assessing each one using specific criteria. Its candidacy for consideration for a geriatric syndrome was assessed based on the quality of data in four domains: multifactorial etiology, shared risk factors with other geriatric syndromes, association with functional decline, and association with increased mortality. The 54 articles reviewed included 24 case series, 13 theoretical articles, 11 observational studies, and six reviews; these were of highly variable methodological quality. The strongest evidence that self-neglect may be a geriatric syndrome includes its often multifactorial etiology, its clear independent association with increased mortality, and the fact that two other geriatric syndromes (cognitive impairment and depression) are risk factors for self-neglect. Self-neglect in older adults is a prevalent problem that appears to have at least some features of a geriatric syndrome. Insofar as the concept of geriatric syndrome has been a useful clinical and research paradigm to create interventions for vulnerable older adults, and no such strategies are available for this vexing and understudied clinical problem, future research is warranted in this area. PMID- 16696753 TI - Ethical issues in mobile psychiatric treatment with homebound elderly patients: the Psychogeriatric Assessment and Treatment in City Housing experience. AB - Unique and challenging ethical difficulties arise during mobile psychiatric treatment of elderly patients. This article outlines and analyzes five of these challenges that have been encountered during nearly 20 years of experience with the Psychogeriatric Assessment and Treatment in City Housing Program in Baltimore, Maryland. The ethical challenges reviewed are: establishing the treatment contract versus the right to refuse treatment, protecting confidentiality versus patient protection, protecting autonomy versus asserting beneficence, treatment termination versus open-ended treatment, and cost versus benefit of care. Ethical challenges with homebound elderly patients are unique because of patient characteristics as well as features of the treatment environment. PMID- 16696754 TI - Interdisciplinary care for older adults with complex needs: American Geriatrics Society position statement. PMID- 16696755 TI - Osteoporosis in older people: a tale of two studies (and three treatments). PMID- 16696756 TI - On the Brazilian elderly Amerindian heritage and the Very Old Italian-Brazilian Longitudinal Study on Aging. PMID- 16696757 TI - Reasons for the high competitiveness in entering a geriatrics fellowship in Brazil: some comparisons with the United States. PMID- 16696760 TI - Rhetoric of research: promoting nutrition products for pressure ulcers. PMID- 16696761 TI - Perioperative haloperidol usage for delirium management. PMID- 16696763 TI - Perioperative haloperidol to prevent postoperative delirium. PMID- 16696765 TI - Vision and ability to take medication. PMID- 16696767 TI - Cognitive function may be important for fall injury prevention trials. PMID- 16696768 TI - Deep vein thrombosis in chronically bedridden elderly individuals. PMID- 16696769 TI - The effects of gum-chewing therapy on salivation in stroke patients. PMID- 16696770 TI - Benefits of combining donepezil plus traditional Japanese herbal medicine on cognition and brain perfusion in Alzheimer's disease: a 12-week observer-blind, donepezil monotherapy controlled trial. PMID- 16696771 TI - Rectal sheath hematoma in an elderly woman after anticoagulation treatment. PMID- 16696772 TI - Gatifloxacin-induced delirium and psychosis in an elderly demented woman. PMID- 16696773 TI - Hyponatremia after fluorescein angiography. PMID- 16696774 TI - Multidisciplinary congestive heart failure clinic for older women: a randomized, controlled trial. PMID- 16696775 TI - Leukocytosis in obese individuals: possible link in patients with unexplained persistent neutrophilia. AB - BACKGROUND: Recently, it was shown that fat tissue produces and releases inflammatory cytokines, and that obesity may be regarded as a state of low-grade inflammation. In this regard, we aimed to establish an association between obesity and persistent leukocytosis. PATIENTS AND METHODS: We present clinical observations of obese subjects primarily referred for further evaluation of leukocytosis without a cause and validated the link between leukocytosis and elevated body mass index (BMI) in a cross-sectional study. RESULTS: During 1999 2005, 327 patients were referred for further investigation because of persistent leukocytosis. Of these, 15.3% were asymptomatic obese, mostly females, with mild persistent neutrophilia accompanied by elevated acute-phase reactants. After careful evaluation, no recognized cause for leukocytosis was found other than the fact that the patients were obese. During a mean follow-up of 45.6 months, the leukocytosis and the elevated acute-phase reactants persisted and no new causes for leukocytosis were evident. Furthermore, in a cross-sectional analysis of 3716 non-smoker subjects, 62 were found to have leukocytosis. Compared with the population with a normal white blood count range, these subjects with leukocytosis had higher BMI, serum C-reactive protein (CRP) levels, waist circumference, and neutrophil and platelet count (all P < 0.0005). After logistic regression analysis, only BMI was shown to be associated with leukocytosis (P < 0.0005). CONCLUSIONS: Obesity is recognized as a possible cause for reactive leukocytosis. Awareness of this 'obesity-associated leukocytosis' may help the clinician to avoid more extensive and unnecessary diagnostic work-up, particularly in similar obese subjects. PMID- 16696776 TI - Clipping for upper gastrointestinal bleeding. PMID- 16696777 TI - Being prepared before the storm. PMID- 16696778 TI - Bioterrorism and natural disasters: the impact of public health law on physicians in the case of an emergency. PMID- 16696779 TI - Military surgery. PMID- 16696780 TI - Pathogenesis and classification of cancer around the gastroesophageal junction- not so different in Japan. AB - Japanese patients with early adenocarcinoma of the esophagus have well-preserved gastric acid secretion, consistent with other parts of the world, reinforcing the likely importance of the luminal environment for pathogenesis. Progress in researching carcinomas that occur around the gastroesophageal junction would be facilitated by consensus-based review of the inadequate approaches currently used for classification of these tumors with subsequent development and widespread adoption of improved criteria. PMID- 16696781 TI - A meta-analysis of the yield of capsule endoscopy compared to other diagnostic modalities in patients with non-stricturing small bowel Crohn's disease. AB - OBJECTIVES: Capsule endoscopy (CE) allows for direct evaluation of the small bowel mucosa in patients with Crohn's disease (CD). A number of studies have revealed significantly improved yield for CE over other modalities for the diagnosis of CD, but as sample sizes have been small, the true degree of benefit is uncertain. Additionally, it is not clear whether patients with a suspected initial presentation of CD and those with suspected recurrent disease are equally likely to benefit from CE. The aim of this study was to evaluate the yield of CE compared with other modalities in symptomatic patients with suspected or established CD using meta-analysis. METHODS: We performed a recursive literature search of prospective studies comparing the yield of CE to other modalities in patients with suspected or established CD. Data on yield among various modalities were extracted, pooled, and analyzed. Incremental yield (IY) (yield of CE--yield of comparative modality) and 95% confidence intervals (95% CI) of CE over comparative modalities were calculated. Subanalyses of patients with a suspected initial presentation of CD and those with suspected recurrent disease were also performed. RESULTS: Nine studies (n = 250) compared the yield of CE with small bowel barium radiography for the diagnosis of CD. The yield for CE versus barium radiography for all patients was 63% and 23%, respectively (IY = 40%, p < 0.001, 95% CI = 28-51%). Four trials compared the yield of CE to colonoscopy with ileoscopy (n = 114). The yield for CE versus ileoscopy for all patients was 61% and 46%, respectively (IY = 15%, p= 0.02, 95% CI = 2-27%). Three studies compared the yield of CE to computed tomography (CT) enterography/CT enteroclysis (n = 93). The yield for CE versus CT for all patients was 69% and 30%, respectively (IY = 38%, p= 0.001, 95% CI = 15-60%). Two trials compared CE to push enteroscopy (IY = 38%, p < 0.001, 95% CI = 26-50%) and one trial compared CE to small bowel magnetic resonance imaging (MRI) (IY = 22%, p= 0.16, 95% CI =-9% to 53%). Subanalysis of patients with a suspected initial presentation of CD showed no statistically significant difference between the yield of CE and barium radiography (p= 0.09), colonoscopy with ileoscopy (p= 0.48), CT enterography (p= 0.07), or push enteroscopy (p= 0.51). Subanalysis of patients with established CD with suspected small bowel recurrence revealed a statistically significant difference in yield in favor of CE compared with all other modalities (barium radiography (p < 0.001), colonoscopy with ileoscopy (p= 0.002), CT enterography (p < 0.001), and push enteroscopy (p < 0.001)). CONCLUSIONS: In study populations, CE is superior to all other modalities for diagnosing non stricturing small bowel CD, with a number needed to test (NNT) of 3 to yield one additional diagnosis of CD over small bowel barium radiography and NNT = 7 over colonoscopy with ileoscopy. These results are due to a highly significant IY with CE over all other modalities in patients with established non-stricturing CD being evaluated for a small bowel recurrence. While there was no significant difference seen between CE and alternate modalities for diagnosing small bowel CD in patients with a suspected initial presentation of CD, the trend toward significance for a number of modalities suggests the possibility of a type II error. Larger studies are needed to better establish the role of CE for diagnosing small bowel CD in patients with a suspected initial presentation of CD. PMID- 16696782 TI - Sensitivity-specificity trade-off for capsule endoscopy in IBD: is it worth it? AB - There are a growing number of reports comparing wireless capsule endoscopy (CE) to other diagnostic tests for use in patients with Crohn's disease. A meta analysis of these studies in this issue of the Journal showed that the "diagnostic yield" of detecting lesions suggestive of active Crohn's disease is much higher with CE compared to other tests. Routine use of CE in Crohn's disease will increase true-positive testing, but also will increase the number of false positive tests. Since the penalty of false-positive testing is very high (unnecessary use of toxic and expensive medications, and the stigma of being diagnosed with a serious chronic disease), CE should be used only in limited clinical situations. In particular, CE may be helpful in Crohn's disease patients without a stricture whose physician needs an added measure of certainty that active disease is present. PMID- 16696783 TI - A population-based case control study of potential risk factors for IBD. AB - BACKGROUND: We aimed to pursue potential etiological clues to Crohn's disease (CD) and ulcerative colitis (UC) through a population-based case control survey study. METHODS: Cases with CD (n = 364) and UC (n = 217), ages 18-50 yr were drawn from the population-based University of Manitoba IBD Research Registry. Potential control subjects were drawn from the population-based Manitoba Health Registry by age, gender, and geographic residence matching to the cases (n = 433). Subjects were administered a multiitem questionnaire. RESULTS: By univariate analysis, some of the variables predictive of CD included lower likelihood of living on a farm, of having drunk unpasteurized milk or having eaten pork, and UC patients were less likely to have drunk unpasteurized milk and to have eaten pork. On multivariate analysis, variables significantly associated with CD were being Jewish (OR = 4.32, 95% CI 1.10-16.9), having a first degree relative with IBD (OR = 3.07, 95% CI 1.73-5.46), ever having smoked (OR = 1.54, 95% CI, 1.06-2.25), living longer with a smoker (OR = 1.03, 95% CI, 1.01-1.04). Being a first generation Canadian (OR = 0.33, 95% CI, 0.17-0.62), having pet cats before age 5 (OR = 0.66, 95% CI, 0.46-0.96) and having larger families (OR = 0.87, 95% CI, 0.79-0.96) were protective against CD. For UC being Jewish (OR = 7.46, 95% CI, 2.33-23.89), having a relative with IBD (OR = 2.23, 95% CI, 1.27 3.9), and ever smoking (OR = 1.62, 95% CI, 1.14-2.32) were predictive. CONCLUSION: This study reinforced the increased risk associated with family history, being Jewish, and smoking history, however, a number of significant associations with CD and UC on univariate and multivariate analysis may support the "hygiene hypothesis" and warrant further exploration in prospective studies. PMID- 16696784 TI - True or false? The hygiene hypothesis for Crohn's disease. AB - The "hygiene hypothesis" for Crohn's disease postulates that multiple childhood exposures to enteric pathogens protect an individual from developing Crohn's disease later in life, while individuals raised in a more sanitary environment are more likely to develop Crohn's disease. In this issue of the American Journal of Gastroenterology, two Canadian case-control studies come to diametrically opposed conclusions regarding the hygiene hypothesis for Crohn's disease. This difference may be partially related to differences in study population (population based vs hospital based), age of onset, different genetic determinants, urban/rural residence (40% rural vs principally urban), or different exposures from the putative causative agent. As of now, the veracity of the hygiene hypothesis for Crohn's disease is not confirmed. PMID- 16696786 TI - Measuring successful treatment of irritable bowel syndrome: is "satisfactory relief " enough? AB - The treatment options for the irritable bowel syndrome (IBS) are expanding as new therapies, including probiotics and serotonin receptor agents, become available. Before any new agents gain widespread use, they must be studied in appropriately designed clinical trials. Symptom improvement remains the key clinically but the best technique to measure symptom improvement is unclear. Many IBS therapy studies have used a binary endpoint such as "Have you had satisfactory relief of your IBS symptoms in the past week? Yes/No?" The study by Whitehead and colleagues in this issue suggests that "satisfactory relief" is affected by baseline symptom severity and may not always truly reflect the symptom burden. Future research needs to determine whether "satisfactory relief" is truly adequate, or whether alternatives such as the proportion of patients achieving a > or = 50% reduction in symptom severity would represent a superior approach to capture clinically important improvement. PMID- 16696785 TI - Inflammatory bowel disease characteristics among African Americans, Hispanics, and non-Hispanic Whites: characterization of a large North American cohort. AB - OBJECTIVES: Inflammatory bowel disease (IBD), comprising primarily of Crohn's disease (CD) and ulcerative colitis (UC), is increasingly prevalent in racial and ethnic minorities. This study was undertaken to characterize racial differences in disease phenotype in a predominantly adult population. METHODS: Phenotype data on 830 non-Hispanic white, 127 non-Hispanic African American, and 169 Hispanic IBD patients, recruited from six academic centers, were abstracted from medical records and compiled in the NIDDK-IBD Genetics Consortium repository. We characterized racial differences in family history, disease location and behavior, surgical history, and extraintestinal manifestations (EIMs) using standardized definitions. RESULTS: African American CD patients were more likely than whites to develop esophagogastroduodenal CD (OR = 2.8; 95% CI: 1.4-5.5), colorectal disease (OR = 1.9; 95% CI: 1.1-3.4), perianal disease (OR = 1.7; 95% CI: 1.03-2.8), but less likely to have ileal involvement (OR = 0.55; 95% CI: 0.32 0.96). They were also at higher risk for uveitis (OR = 5.5; 95% CI: 2.3-13.0) and sacroiliitis (OR = 4.0; 95% CI: 1.55-10.1). Hispanics had higher prevalence of perianal CD (OR = 2.9; 95% CI: 1.8-4.6) and erythema nodosum (3.3; 95% CI: 1.7 6.4). Among UC patients, Hispanics had more proximal disease extent. Both African American and Hispanic CD patients, but not UC patients, had lower prevalences of family history of IBD than their white counterparts. CONCLUSIONS: There are racial differences in IBD family history, disease location, and EIMs that may reflect underlying genetic variations and have important implications for diagnosis and management of disease. These findings underscore the need for further studies in minority populations. PMID- 16696787 TI - Alosetron: ischemic colitis and serious complications of constipation. AB - Drugs such as alosetron that modulate serotonin effects by stimulating or blocking its receptors may play an important role in the treatment of some patients with irritable bowel system. In the case of alosetron, a 5HT-3 antagonist, an analysis of data from randomized clinical trials and postmarketing experiences have demonstrated a causal relationship between this drug and ischemic colitis and serious complications of constipation. Because the mechanism(s) of drug-induced ischemic colitis and possibly other forms of intestinal ischemia associated with alosetron have not been elucidated, there is need to further assess risk with regard to patient susceptibility and other factors. PMID- 16696788 TI - Clinical performance of original and revised Bethesda guidelines for the identification of MSH2/MLH1 gene carriers in patients with newly diagnosed colorectal cancer: proposal of a new and simpler set of recommendations. AB - Identification of individuals who should undergo hereditary nonpolyposis colorectal cancer (HNPCC) genetic testing is a critical and difficult issue. For this purpose, the National Cancer Institute outlined a set of recommendations, the Bethesda guidelines, which have recently been revised. OBJECTIVE: To compare the clinical performance of original and revised Bethesda guidelines for the detection of MSH2/MLH1 gene carriers in patients with colorectal cancer. METHODS: A total of 1,222 patients with newly diagnosed colorectal cancer were included in the EPICOLON study, a prospective, multicenter, nationwide epidemiology survey aimed at establishing the incidence of HNPCC in Spain (JAMA 2005; 293:1986-1994). Performance characteristics of the original and revised Bethesda guidelines were assessed with respect to the presence of MSH2/MLH1 germline mutations. Logistic regression analysis was performed to establish the most effective strategy. RESULTS: Original or revised Bethesda guidelines were equivalent strategies in terms of sensitivity (100%vs 100%; ns), specificity (98.1%vs 97.9%; ns), and overall accuracy (98.1%vs 97.9%; ns), as well as positive (25.8%vs 24.2%) and negative predictive values (100%vs 100%). The most discriminating individual variables were criteria number 1 (i.e., fulfillment of the Amsterdam criteria; RR = 34.14; 95% CI = 6.85-170.16; p < 0.001) and number 2 (i.e., individuals with two HNPCC-related neoplasms; RR = 35.63; 95% CI = 4.83-262.6; p < 0.001) of the original guidelines, and criterion number 1 of the revised guidelines (i.e., colorectal cancer diagnosed under 50 yr of age; RR = 29.34; 95% CI = 3.81-225.96; p= 0.001). The aggregation of these three criteria was equivalent to both Bethesda guidelines in terms of sensitivity (100%) and negative predictive value (100%), but superior to the revised criteria regarding specificity (98.5%; p < 0.05), overall accuracy (98.5%; p < 0.05), and positive predictive value (30.8%). CONCLUSIONS: Original and revised Bethesda guidelines are equivalent, highly effective criteria for the identification of MSH2/MLH1 gene mutation carriers in patients with newly diagnosed colorectal cancer. A new set of recommendations, based on a combination of some of their individual criteria, may provide additional advantages in terms of effectiveness. PMID- 16696789 TI - Gastroparesis: clinical update. AB - Gastroparesis refers to chronically abnormal gastric motility characterized by symptoms suggestive of mechanical obstruction and delayed gastric emptying in the absence of mechanical obstruction. It may be idiopathic or attributable to neuropathic or myopathic abnormalities, such as diabetes mellitus, postvagotomy, postviral infection, and scleroderma. Dietary and behavioral modification, prokinetic drugs, and surgical interventions have been used in managing patients with gastroparesis. Although mild gastroparesis is usually well managed with these treatment options, severe gastroparesis may be very difficult to control and may require referral to a specialist center if symptoms are intractable despite pharmacological therapy and dietetic support. New advances in drug therapy, botulinum toxin injection, and gastric electrical stimulation techniques have been introduced and might provide new hope to patients with refractory gastroparesis. This article critically reviews the advances in the field from the perspective of the clinician. PMID- 16696790 TI - Rectal hyposensitivity. AB - Rectal hyposensitivity (RH) relates to a diminished perception of rectal distension that is diagnosed during anorectal physiologic investigation. There have been few direct studies of this physiologic abnormality, and its contribution to the development of functional bowel disorders has been relatively neglected. However, it appears to be common in patients with such disorders, being most prevalent in patients with functional constipation with or without fecal incontinence. Indeed, it may be important in the etiology of symptoms in certain patients, given that it is the only "apparent" identifiable abnormality on physiologic testing. Currently, it is usually diagnosed on the basis of elevated sensory threshold volumes during balloon distension in clinical practice, although such a diagnosis may be susceptible to misinterpretation in the presence of altered rectal wall properties, and thus it is uncertain whether a diagnosis of RH reflects true impairment of afferent nerve function. Furthermore, the etiology of RH is unclear, although there is limited evidence to support the role of pelvic nerve injury and abnormal toilet behavior. The optimum treatment of patients with RH is yet to be established. The majority are managed symptomatically, although "sensory-retraining biofeedback" appears to be the most effective treatment, at least in the short term, and is associated with objective improvement in the rectal sensory function. Currently, fundamental questions relating to the contribution of this physiologic abnormality to the development of functional bowel disorders remain unanswered. Acknowledgment of the potential importance of RH is thus required by clinicians and researchers to determine its relevance. PMID- 16696792 TI - Endoscopic mucosal resection for Paris type II colorectal neoplasia: hemostatic complications and resection margin histopathological assessment using coagulation current without routine endoclip closure. PMID- 16696793 TI - Critical appraisal of individual trials? PMID- 16696794 TI - Mycobacterium avium subspecies paratuberculosis: a human pathogen causing most cases of Crohn's disease. PMID- 16696798 TI - Review article: a critical view on impaired accommodation as therapeutic target for functional dyspepsia. AB - Several important pathophysiological mechanisms have been identified in functional dyspepsia, however a complete understanding of these mechanisms and beneficial therapeutic strategies are still lacking. Based on the currently available literature we aimed at providing a critical view on one of these pathophysiological mechanisms, impaired accommodation. Although impaired gastric accommodation is identified as a major pathophysiological mechanism, the clinical evidence supporting its role as an important therapeutic target is currently still lacking. Treatment with fundic relaxant drugs has shown conflicting results and has been rather disappointing in general. These negative findings could be explained by the fact that impaired fundic accommodation is part of a more complex disorder involving other regions of the proximal gut or by the increasing insight that central mechanisms may play an important role. Future studies of impaired accommodation should take these considerations into account. PMID- 16696799 TI - Systematic review: adipose tissue, obesity and gastrointestinal diseases. AB - BACKGROUND: Obesity is increasingly being recognized as a risk factor for a number of benign and malignant gastrointestinal conditions. However, literature on the underlying pathophysiological mechanisms is sparse and ambiguous. Insulin resistance is the most widely accepted link between obesity and disease, particularly colorectal cancer. The recognition that intra-abdominal fat is immunologically active sheds new light not only on the pathogenesis of obesity related gastrointestinal conditions, but also on inflammatory conditions such as Crohn's disease. AIM: To describe the biology of adipose tissue, its impact on the immune system and explores the possible underlying mechanisms linking obesity to gastrointestinal diseases. It also looks at the role of mesenteric fat in determining severity and course of Crohn's disease. METHODS: Relevant English language literature and abstracts cited on MEDLINE database were reviewed. RESULTS: Our recent finding of an association between obesity and subclinical bowel inflammation suggests that, apart from promoting generalized immune activation, fat also evokes local immune responses. We propose that the proinflammatory milieu promoted by obesity could underlie many of these associations and that the mechanism implicating insulin resistance may merely represent an epiphenomenon. In Crohn's disease, on the other hand, intra abdominal fat may provide a protective mechanism. CONCLUSION: The potential of adipose tissue as a therapeutic target is vast and needs exploration. PMID- 16696800 TI - Systematic review: microscopic colitis. AB - BACKGROUND: Collagenous and lymphocytic colitis are fairly common causes of chronic non-bloody diarrhoea, especially in elderly female. AIM: To present a systematic review of microscopic colitis. METHODS: A PubMed search using the MeSH terms microscopic colitis, collagenous colitis, lymphocytic colitis and chronic diarrhoea was performed. RESULTS: Annual incidence of each disorder is 4 6/100,000 inhabitants. The aetiology is unknown. Clinical characteristics are well described and there is an association with autoimmune diseases. Budesonide is the best-documented short-term treatment of collagenous colitis. In meta analysis pooled odds ratio for clinical response after 6-8 weeks of treatment was 12.3 (95% CI: 5.5-27.5) in comparison with placebo. The evidence for bismuth subsalicylate is weaker and the effectiveness of other alternatives such as loperamide, cholestyramine, aminosalicylates, probiotics, or Boswellia serrata extract is unknown. Although unproven, in unresponsive severe disease azathioprine or methotrexate may be tried. No controlled trials have been carried out in lymphocytic colitis. The long-term prognosis of microscopic colitis is good, serious complications are rare and there is no increased mortality. CONCLUSIONS: Clinical and epidemiological aspects of microscopic colitis are well described. Budesonide is the best-documented short-term therapy in collagenous colitis, but the optimal long-term strategy needs further study. Controlled treatment data of lymphocytic colitis are awaited for. PMID- 16696801 TI - Systematic review: evidence-based management of hepatocellular carcinoma--an updated analysis of randomized controlled trials. AB - The treatment strategy of hepatocellular carcinoma applied following scientific guidelines is only supported by 77 randomized controlled trials published so far, a figure that clearly pinpoints hepatocellular carcinoma as an 'orphan' cancer in terms of clinical research when compared with other high-prevalent cancers worldwide. A systematic review analysing 61 randomized controlled trials (1978 2002) showed a modest survival benefit from chemoembolization in patients with intermediate tumours, and the lack of an effective first-line treatment option for patients with advanced disease. These conclusions have been endorsed by the European Association for the Study of the Liver and the American Association for the Study of Liver Diseases. The present updated evidence-based approach includes 16 randomized controlled trials published from 2002 to 2005 assessing percutaneous ablation (seven), other loco-regional therapies (three) and systemic therapies (six). Eight showed high-quality methodological profiles. Four randomized controlled trials demonstrated a better local hepatocellular carcinoma control in tumours larger than 2 cm treated by radiofrequency ablation compared with ethanol injection. No survival advantages were obtained from systemic treatments in patients with advanced hepatocellular carcinoma, an area that is an unmet need. Therefore, there is an urgent request to conduct well-designed phase III investigations in hepatocellular carcinoma patients. PMID- 16696802 TI - Personal view: current role of artificial liver support devices. AB - Enthusiasm for liver support devices, particularly cell-based biological systems and albumin dialysis, increased over the last decade and there has been considerable clinical activity both within and without the construct of clinical trials. Most data have been generated on patients with acute liver failure or in patients with decompensation of chronic liver disease, often referred to as acute on-chronic liver failure. In acute liver failure liver, liver support devices are more realistically being used as a 'bridge' to liver transplantation rather than to transplant-free survival. In acute-on-chronic liver failure the clinical objective of attaining clinical stability with treatment appears more achievable. The so-called bioartificial liver device, based on porcine hepatocytes, is the most extensively evaluated biological device. A sizeable clinical trial failed to demonstrate efficacy, but secondary analyses suggest it would be unwise to assume futility had been established with this device. Molecular adsorbent recirculating system leads the way in the non-biological category in terms of the number of patients treated, but data from large clinical trials are not yet available. One of the strongest conclusions of this review is that the amount of high-quality data available on liver support devices dramatically understates the effort and money that have been expended in their assessment. It is very clear that randomized controlled trials are mandatory to establish clinical efficacy, but it is less clear how the ideal trial should be constructed. PMID- 16696803 TI - Meta-analysis: the adjuvant role of thymopentin on immunological response to hepatitis B virus vaccine in end-stage renal disease. AB - BACKGROUND: It has been calculated that 30-40% of dialysis patients fail to produce antibodies to HBsAg antigen after vaccination towards hepatitis B virus. Several authors have reported on the benefit of thymopentin (TP5) as adjuvant to vaccine against hepatitis B virus in patients receiving regular dialysis. However, consistent information on this issue is still lacking. AIMS: To evaluate efficacy and safety of thymopentin as adjuvant to hepatitis B vaccine in dialysis patients by performing a systematic review with a meta-analysis of clinical trials. METHODS: We used the random effects model of DerSimonian and Laird, with heterogeneity and sensitivity analyses. RESULTS: We identified 11 studies involving 272 unique patients with end-stage renal disease. Only prospective, controlled trials were included. Pooling of study results did not show a significant increase in seroresponse rate among study (thymopentin plus hepatitis B virus vaccine) vs. control (hepatitis B virus vaccine alone) patients; the pooled odds ratio of failure to respond to hepatitis B virus vaccine was 0.677 (95% confidence intervals: 0.285-1.605); no heterogeneity was found (P = 0.0001). Thymopentin significantly improved the seroresponse rate in the subgroup of trials based on greater thymopentin doses (OR: 0.184; 95% CI: 0.085-0.398). CONCLUSIONS: Our meta-analysis showed that thymopentin significantly improved the seroresponse rate towards hepatitis B vaccine only in dialysis patients treated with higher thymopentin doses. The limited number of patients precluded definitive conclusions. PMID- 16696804 TI - Efficacy of Lactobacillus GG in maintaining remission of ulcerative colitis. AB - BACKGROUND: Aminosalicylates are the mainstay of therapy to prevent relapse of quiescent ulcerative colitis. The rationale for using probiotics is based on the evidence implicating intestinal bacteria in the pathogenesis of this disorder. AIM: To evaluate the efficacy of Lactobacillus GG alone or in combination with mesalazine vs. mesalazine as maintenance treatment in ulcerative colitis. PATIENTS AND METHODS: 187 ulcerative colitis patients with quiescent disease were randomized to receive Lactobacillus GG 18 x 10(9) viable bacteria/day (65 patients), mesalazine 2400 mg/day (60 patients) or Lactobacillus GG + mesalazine (62 patients). Disease activity index, endoscopic and histological scores were determined at 0, 6 and 12 months and in case of relapse. The primary end point was to evaluate sustained remission. RESULTS: Overall analysis showed no difference in relapse rate at 6 (P = 0.44) and 12 months (P = 0.77) among the three treatment groups. However, the treatment with Lactobacillus GG seems to be more effective than standard treatment with mesalazine in prolonging the relapse free time (P < 0.05). CONCLUSIONS: Lactobacillus GG seems to be effective and safe for maintaining remission in patients with ulcerative colitis, and it could represent a good therapeutic option for preventing relapse in this group of patients. PMID- 16696805 TI - Treatment of haemorrhagic radiation-induced proctopathy using small volume topical formalin instillation. AB - BACKGROUND: Between 2 and 5% of patients undergoing pelvic radiotherapy develop chronic radiation proctopathy, occurring as a result of damage to the rectal mucosa during the treatment. Rectal bleeding of varying severity can occur as a consequence. There have been no formal trials of treatment for haemorrhagic radiation proctopathy and a variety of methods are currently used. AIM: In a retrospective study of 20 patients treated at a single centre, we assessed the efficacy of small volume topical formalin instillation to control bleeding from radiation proctopathy. METHOD: Patients were treated by a single operator using 20 mL of a 5% solution of formalin instilled into the rectum via a flexible sigmoidoscope for 3 min. Patients were followed up for an average of 22.7 months (range: 2-69). RESULTS: A single session of formalin treatment was effective in 13 of 20 (65%) patients and a further four (20%) patients responded to a second treatment. No complications of the treatment was identified. CONCLUSION: Small volume formalin instillation therapy appears to be safe and effective in the context of haemorrhagic radiation proctopathy. The technique is simple, inexpensive, quick and requires no sedation. We suggest that it should be considered as a first line for patients presenting with this distressing condition. PMID- 16696806 TI - White wine and beer induce gastro-oesophageal reflux in patients with reflux disease. AB - BACKGROUND: An induction of gastro-oesophageal reflux has been reported after ingestion of alcoholic beverages in healthy volunteers. However, it is unknown whether reflux in gastro-oesophageal reflux disease patients will be enhanced by the ingestion of alcoholic beverages. AIM: To investigate the effects of wine and beer on postprandial reflux in reflux patients. METHODS: Twenty-five patients (reflux oesophagitis 15, non-erosive reflux disease 10; 18 men and seven women) drank 300-mL white wine (n = 17), 500-mL beer (n = 8), or identical amounts of tap water (controls) together with a standardized meal in a randomized order. pH measurement was carried out during three postprandial hours by pH-metry and the percentage of time pH < 4 was calculated. RESULTS: Both alcoholic beverages increased reflux compared with water [wine 23% (median), water 12%, P < 0.01; beer 25%, water 11%, P < 0.05]. Between wine and beer, no difference in reflux induction was obtained. The reflux induction was seen in patients with (23%, P < 0.01) and without reflux oesophagitis (22%, P < 0.05) and in both sexes (women 23%, men 25%, P < 0.05 each). CONCLUSIONS: Ingestion of commonly consumed alcoholic beverages such as wine and beer induces gastro-oesophageal reflux in gastro-oesophageal reflux disease patients. Therefore, these patients should be advised to avoid the intake of large amounts (> or = 300 mL) of these beverages. PMID- 16696807 TI - Declining incidence of peptic ulcer but not of its complications: a nation-wide study in The Netherlands. AB - BACKGROUND: Despite reports of decreasing hospitalizations and mortality due to peptic ulcer, it is unclear whether the incidence has truly declined over time. AIM: To investigate time trends in the incidence of and in hospital admission rates for peptic ulcer in the Netherlands. METHODS: The nationwide registry of pathology reports (PALGA) and the national registry of hospital admissions (Landelijke Medische Registratie) were used. Standardized morbidity ratios were calculated to assess the magnitude of the changes. RESULTS: The age-adjusted incidence of gastric ulcer halved for both men (standardized morbidity ratio 0.48; CI 0.46-0.49) and women (standardized morbidity ratio 0.49; CI 0.47-0.51). Although the number of gastric biopsies obtained at endoscopy increased, the proportion with a diagnosis of peptic ulcer decreased by more than 50% (standardized morbidity ratio 0.47; CI 0.46-0.49). The admission rate for peptic ulcer more than halved between 1980 and 2003. In contrast, admission rates for complicated ulcers barely changed and slightly increased among women. CONCLUSIONS: The incidence of histopathologically confirmed gastric ulcer halved between 1992 and 2003 in the Netherlands. As the number of gastric biopsies increased in this period, a true decrease is likely. Hospital admissions for peptic ulcer declined dramatically between 1980 and 2003, but remained unchanged or slightly increased for complicated ulcers. PMID- 16696808 TI - Switch patterns before and after patent expiry of omeprazole: a case study in The Netherlands. AB - BACKGROUND: An increase of therapeutic substitution after patent expiry might have a negative effect on cost-savings generated with newly introduced generic drugs. To evaluate influences of patent expiry on therapeutic substitution, switch behaviour before and after patent expiry was investigated. AIM: To describe proton pump inhibitor use and investigate substitution patterns from omeprazole before and after patent expiry. METHODS: Data were obtained from the InterAction DataBase. Proportional proton-pump inhibitor use was identified per quarter during the study period 2000-2003. For the second part two cohorts--one before and one after patent expiry--were defined. The number of switchers was quarterly identified during 2-year follow-up period. For statistical analyses the chi-square test and hazard ratio were used. RESULTS: In proportional use, a downward trend for omeprazole was found. After patent expiry, significantly more patients switched to other proton pump inhibitors (P < 0.001). The hazard ratio of 0.62 (95% CI: 0.57-0.69), indicates that for every six patients switching before patent expiry, 10 patients switch after patent expiry. CONCLUSION: After patent expiry more patients switch to another proton pump inhibitor. In light of the total savings achieved with generic omeprazole, the importance of this negative impact on total cost-savings on proton pump inhibitors is unclear. PMID- 16696809 TI - Prevalence and sociodemographics of reflux symptoms in Germany--results from a national survey. AB - BACKGROUND: Although gastro-oesophageal reflux disease is a common gastrointestinal disorder, the prevalence rates differ depending on the population studied. AIM: To determine the prevalence of reflux symptoms in the German adult population and any associations with age, gender or socio-economic status. METHODS: A total of 7124 subjects were interviewed as part of the German National Health Interview and Examination Survey, which is a representative sample of the general adult population. The prevalence of mild, moderate or severe heartburn or regurgitation was calculated. Logistic regression models were developed to determine the relationship between reflux symptoms and sociodemographic factors such as age, gender and socio-economic status. RESULTS: The overall prevalence of moderate and severe reflux symptoms was 14% and 4%, respectively. The prevalence increased with age and was highest in the age group 60-69 years with 25% of these subjects reporting moderate or severe reflux symptoms. Being male, it increased the risk of mild, but not of moderate or severe symptoms. Lower socio-economic status was associated with moderate or severe symptoms. CONCLUSIONS: Because of their high prevalence, reflux symptoms are of major public health importance. PMID- 16696810 TI - Effect of frequent dosing of an oral proton pump inhibitor on intragastric pH. AB - BACKGROUND: Treatment with a continuous i.v. proton pump inhibitor is presumed to promote clot formation and stability by sustaining intragastric pH > or = 6. AIM: We postulated that very frequent oral dosing of proton pump inhibitors should simulate i.v. infusion and achieve similar pH control. METHODS: Twenty healthy volunteers were stratified by Helicobacter pylori status (10 positive; 10 negative) and had determination of CYP2C19 status. After an overnight fast, an intragastric pH probe was placed. Subjects received 120 mg of lansoprazole at 8 am and 30 mg every 3 h until 8 pm. Intragastric pH was measured over 24 h, and lansoprazole plasma concentrations were determined at five time points. RESULTS: Intragastric pH was > or = 6 for 41% (95% CI: 30-53%) of the 15-h period from 8 am-11 pm and 46% (95% CI: 35-56%) of the 24-h period (8-8 am). The mean proportion of patients with pH > or = 6 was not significantly different in H. pylori-positive vs. negative patients. Only 25% of subjects sustained pH > or = 6 for at least 60% of the 15-h period, and 35% had a sustained pH > or = 6 for at least 60% of the 24-h period. CONCLUSIONS: A dose of 120 mg of oral lansoprazole followed by standard 30 mg doses of lansoprazole every 3 h did not reliably sustain pH at the desired level of 6. PMID- 16696811 TI - Treatment with botulinum toxin of octo-nonagerians with oesophageal achalasia: a two-year follow-up study. AB - BACKGROUND: Treatment of oesophageal achalasia with intrasphincteric injections of botulinum toxin has proved to be a successful alternative treatment modality. However, little is known about its long-term effects in very old patients. AIM: To evaluate the effects of such treatment in octo-nonagerians during a 2-year follow-up period. PATIENTS AND METHODS: Thirty-three patients with idiopathic oesophageal achalasia (range 81-94 years) entered the study. After basal evaluation and screening procedures, 100 U of botulinum toxin was injected at the lower oesophageal sphincter, and the procedure was repeated 1 month later. Data were collected at baseline and were compared after 1 and 2 years following the procedure. RESULTS: Seventy-eight per cent of patients were considered responders at 1 year and 54% were considered responders at 2 years. The weight gain at the end of the follow-up period was 2 (0-3) kg. No significant relationship was found between baseline lower oesophageal sphincter pressure and symptoms score after 1 and 2 years of follow-up; moreover, no major complications of botulinum toxin therapy were reported. CONCLUSION: Treatment of very old achalasic patients with botulinum toxin is safe, effective and yields good quality of life in a substantial proportion of these subjects. PMID- 16696812 TI - Diagnosis of liver fibrosis by transient elastography (FibroScan) and non invasive methods in Crohn's disease patients treated with methotrexate. AB - BACKGROUND: Methotrexate is an effective treatment in Crohn's disease, which may induce liver fibrosis with high cumulative doses. Transient elastography (FibroScan, Echosens, Paris, France) is a new non-invasive rapid, allowing assessment of liver fibrosis by measurement of liver stiffness. AIM: A prospective study to evaluate liver fibrosis with FibroScan and non-invasive biochemical methods in Crohn's disease patients treated with methotrexate. METHODS: Consecutive Crohn's disease patients had evaluation of liver fibrosis with non-invasive methods. Two subgroups of patients were compared: cumulative dose of methotrexate of more than 1500 mg (group 1) and naive for methotrexate (group 2). Liver biopsy was performed in patients with persistent liver enzyme abnormalities or FibroScan value >8.7 kPa. RESULTS: Fifty-four consecutive Crohn's disease patients were fully investigated (45 females, mean age 41 +/- 14 years). Median FibroScan values were similar in group 1 (n = 21) and in group 2 (n = 33), 5.5 and 4.5 kPa, respectively. FibroScan values were not correlated with the cumulative dose of methotrexate. CONCLUSION: In Crohn's disease patients treated with a high dose of methotrexate, significant liver fibrosis is rare and not accurately detected with liver enzymes abnormalities. FibroScan could be recommended and liver biopsy could be performed only with patients with high values and/or with chronic liver enzymes abnormalities. PMID- 16696813 TI - Determinants of quality of life in chronic liver patients. AB - BACKGROUND AND AIM: Health-related quality of life of patients with chronic liver disease has been shown to be impaired in numerous studies. However, the factors which influence health-related quality of life in treated chronic liver patients are not quite known. This is the first study to assess the impact of physical and psychosocial determinants on a weighted score of health-related quality of life in patients with chronic liver disease. METHODS: The data of 1175 chronic liver patients were used to assess the relationship between items of the disease specific Liver Disease Symptom Index 2.0 and the Short Form (SF)-6D weighted utility score by means of linear regression analyses. RESULTS: Health-related quality of life was most strongly related to disease severity (B = -0.029) and joint pain (B = -0.023). Depression (B = -0.014), pain in the right upper abdomen (B = -0.014), decreased appetite (B = 0.014) and fatigue (B = -0.013) were also strongly related to health-related quality of life. In hepatitis C virus patients, disease severity (B = -0.037) and depression (B = -0.030) were strong determinants of health-related quality of life. CONCLUSIONS: This study shows that health-related quality of life in chronic liver patients is clearly determined by disease severity, joint pain, depression, decreased appetite and fatigue. These patients may benefit most from interventions aimed at improving adaptation to the symptoms described. PMID- 16696814 TI - Serum bilirubin and colorectal cancer risk: a population-based cohort study. AB - BACKGROUND: Bilirubin has antioxidant properties and has been postulated to protect against the development of malignancies. AIM: To investigate whether baseline serum bilirubin concentration predicts the incidence of colorectal cancer in a nationally representative sample of the US population. METHODS: Participants of the first National Health and Nutrition Examination Survey were divided into four groups based on quartiles of baseline serum bilirubin concentration in mg/dL: <0.38 (n = 1410), 0.38 to <0.5 (n = 1287), 0.5 to <0.6 (n = 1048) and > or = 0.6 (n = 1742). The incidence of colorectal cancer during the following 20 years was determined from hospitalization records and death certificates. RESULTS: 110 cases of colorectal cancer-related death or hospitalization were identified among 5487 participants during 88,339 person years of follow-up (12 per 10,000 person-years). There was no association between baseline serum bilirubin concentration and the incidence of colorectal cancer either in unadjusted analyses or after adjusting for age, gender, ethnicity, smoking, body mass index, alcohol consumption and educational attainment. CONCLUSIONS: Baseline serum bilirubin concentration did not predict the subsequent incidence of colorectal cancer in this population-based cohort study. PMID- 16696815 TI - A pilot study of atorvastatin treatment in dyslipemid, non-alcoholic fatty liver patients. AB - BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) encompasses a wide spectrum of liver injury. Currently, there are no proven effective therapies available. Atorvastatin is a new 3-hydroxy-3-metylglutaryl coenzyme A reductase inhibitor that reduces lipid serum levels. AIM: To evaluate the effectiveness and safety of atorvastatin in dyslipemid, non-alcoholic fatty liver patients. PATIENTS AND METHODS: We prospectively enrolled 25 patients with NAFLD; 22 of them completed the study. Body mass index, serum lipids, liver function tests and liver density assessed by echography were measured at baseline and after 1, 3, 6, 9 and 12 months of treatment. Normalization of transaminases and/or improvement in liver density were treatment end points. Patients received atorvastatin (10-80 mg/daily) according to basal serum choleresterol levels; additionally, they were given standard weight-loss counselling and encouraged to follow a low fat diet. RESULTS: All 22 patients (14 men, mean age 47 +/- 10 years) had high cholesterol levels at baseline and 11 (44%) also presented high trygliceride levels. After 6 months of treatment, eight patients (36.3%) presented normal transaminase levels. The remaining patients continued treatment for 12 months when 20% of patients presented with normal transaminase levels, while the other patients showed a 10% reduction in basal levels. Mean cholesterol levels were 268.5 +/- 44.2 and 186.8 +/- 14.4 mg/dL before and after treatment, respectively (P < 0.05). The mean body mass index was 27.4 +/- 3.1 at baseline and 26.3 +/- 2.8 kg/cm2 at the end of treatment (P > 0.05). No side effects were reported. CONCLUSIONS: Serum aminotransferase and lipid levels were reduced significantly in all patients with atorvastatin treatment. Therapy with atorvastatin in NAFLD patients with hyperlipidemia was found to be both effective and safe. PMID- 16696816 TI - Intrahepatic cholestasis of pregnancy: three novel MDR3 gene mutations. AB - BACKGROUND: The aetiology of intrahepatic cholestasis of pregnancy is unknown, but more than 10 different MDR3 gene mutations have recently been identified. AIM: To evaluate the genetic contribution of the MDR3 gene in the pathogenesis of intrahepatic cholestasis of pregnancy in Italian subjects. METHODS: We performed a multicentre prospective case-control study, enrolling 80 women with intrahepatic cholestasis of pregnancy at the third trimester of pregnancy and 80 pregnant women without intrahepatic cholestasis of pregnancy. Genomic DNA was extracted from peripheral venous blood leucocytes using standard procedures. The polymerase chain reaction was used to amplify exon 14 of the MDR3 gene and the polymerase chain reaction products were sequenced using a Big Dye Terminator Cycle Sequencing kit. RESULTS: Three novel non-synonymous heterozygous mutations in exon 14 were found (4%; E528D, R549H, G536R) among the 80 intrahepatic cholestasis of pregnancy patients, whereas the pregnant controls were all negative for exon 14 polymorphisms. The three patients involved had normal GGT and bilirubin, but high levels of both ALT and serum bile acids. One had cholesterol bile stones. The outcome of pregnancy was normal for two (with vaginal delivery), while foetal distress was recorded in the third. CONCLUSIONS: These three novel mutations add further information on the involvement of the MDR3 gene in intrahepatic cholestasis of pregnancy. As in other studies, we found only heterozygous mutations that could cause an impaired transport protein function, not its absence (which is responsible for more severe liver disease). Different genetic backgrounds might justify the presence of novel MDR3 gene mutations. PMID- 16696817 TI - Pilot study of the efficacy of renzapride on gastrointestinal motility and symptoms in patients with constipation-predominant irritable bowel syndrome. AB - AIM: To investigate the efficacy and safety of renzapride, a potent 5 hydroxytryptamine type-4 receptor full agonist and 5-hydroxytryptamine type-3 receptor antagonist in patients with constipation-predominant irritable bowel syndrome. METHODS: In this dose-escalating pilot study, 17 patients with constipation-predominant irritable bowel syndrome received placebo, renzapride 2 mg o.d. and renzapride 2 mg b.d. sequentially for 28 days. Response was determined by radio-opaque marker measurement of overall gastrointestinal and segmental colonic transit and patients' assessment of their irritable bowel syndrome symptoms. RESULTS: Renzapride reduced mean overall gastrointestinal transit time (placebo, 2.9 +/- 1.6 days; renzapride 2 mg o.d., 2.6 +/- 1.4 days; renzapride 2 mg b.d., 1.9 +/- 1.6 days) (P = 0.024) and accelerated segmental colonic transit, with statistically significant differences for renzapride 2 mg b.d. over placebo in caecum/ascending colon (P = 0.019) and descending colon (P = 0.022). Renzapride also reduced abdominal pain, increased the number of pain-free days and improved stool consistency. The frequency of reported adverse events was similar on renzapride and placebo. CONCLUSIONS: Renzapride is well-tolerated, stimulates gastrointestinal transit and improves symptoms in patients with constipation-predominant irritable bowel syndrome, particularly at the 2 mg b.d. dose, where improvements in gastrointestinal symptoms were evident over placebo. This study has established proof of concept and supports further investigation of renzapride in patients with constipation-predominant irritable bowel syndrome. PMID- 16696818 TI - Determinants of healthcare-seeking behaviour among subjects with irritable bowel syndrome. AB - BACKGROUND: Doctor visits for irritable bowel syndrome are associated with high medical costs. Predictors of medical consultation for irritable bowel syndrome remain poorly understood. AIM: To determine factors associated with healthcare seeking for irritable bowel syndrome. METHODS: Subjects from previous US population-based survey were contacted 2 years later. Those who continued to have irritable bowel syndrome were included. RESULTS: 49% of subjects sought medical care for abdominal symptoms in the past year. Healthcare seeking did not differ significantly between males and females, but more females received an irritable bowel syndrome diagnosis. Predictors of irritable bowel syndrome healthcare seeking differed by gender. In multivariate analysis, age > or = 55 years (OR = 2.8, 95% CI: 1.5-5.4), fear abdominal symptoms relates to serious illness (OR = 1.7, 95% CI: 0.95-3.1), decreased bowel movements (OR = 1.8, 95% CI: 0.98-3.2), dyspepsia (OR = 1.7, 95% CI: 0.94-3.2) and pelvic pain (OR = 2.3, 95% CI: 1.2 4.4) were associated with seeking care in females. Among males, being disabled (OR = 11.6, 95% CI: 2.4-56.1) and abdominal cramping (OR =4.3, 95% CI: 1.2-15.4) were associated with seeking care. Healthcare seekers had lower irritable bowel syndrome-related quality of life. Neither pain severity nor mental health status was associated with seeking care. CONCLUSION: Healthcare-seeking behaviour among irritable bowel syndrome patients was determined by presence of comorbidities and extent that irritable bowel syndrome affected quality of life, not physical symptoms or mental health status. PMID- 16696819 TI - Gastric juice--a natural-born killer. PMID- 16696821 TI - Prolonged treatment with neostigmine for resolution of acute colonic pseudo obstruction. PMID- 16696822 TI - Prophylactic treatment of bipolar disorder in pregnancy and breastfeeding: focus on emerging mood stabilizers. AB - OBJECTIVES: Bipolar disorders are reported to have a high incidence during childbearing years and the need may arise to start or continue a pharmacological treatment during pregnancy and the postpartum period. In the last few years several investigations have evaluated the efficacy of emerging mood-stabilizing agents in the treatment of bipolar disorders, such as lamotrigine, olanzapine, risperidone, quetiapine, aripiprazole and ziprasidone. A number of studies, which examined the use of oxcarbazepine, point to its potential usefulness in prophylactic treatment. The aim of this review is to compare information from the literature on the safety of lamotrigine, oxcarbazepine, risperidone, olanzapine, and quetiapine to the safety data on classic mood stabilizers during pregnancy and the postpartum period. METHODS: A computerized search carried out from 1980 to April 5, 2006 led to the summarization of the results. (References were updated after acceptance and prior to publication.) RESULTS: Emerging mood stabilizers show uncertain safety parameters in pregnancy and lactation. Limited information on lamotrigine and oxcarbazepine does not suggest a clear increase in teratogenicity, while olanzapine appears to be associated with a higher risk of metabolic complications in pregnant women. Data about risperidone and quetiapine are still inconclusive. Finally, the literature on the safety of these compounds in breastfeeding is anecdotal. CONCLUSIONS: Untreated pregnant bipolar women are at an increased risk of poor obstetrical outcomes and relapse of affective symptoms. On the other hand, classic antiepileptic drugs are well-known human teratogens, whereas data on lithium are partially ambiguous. The safety of emerging mood stabilizers in pregnancy and breastfeeding has not been examined extensively. Therefore, when approaching bipolar disorder, if possible, each episode must be considered separately. PMID- 16696823 TI - Symptomatic and functional outcome 12 months after a first episode of psychotic mania: barriers to recovery in a catchment area sample. AB - OBJECTIVE: Recent studies have shown that outcome in mania is worse than previously thought. Such studies have been conducted in selected samples with restrictive measures of outcome. We aimed to explore outcome and its predictors in a catchment area sample of first-episode psychotic mania of DSM-III-R bipolar I disorder. METHODS: Prospective 6 and 12 months follow-up was conducted with 87 DSM-III-R first-episode psychotic mania patients admitted to Early Psychosis Prevention and Intervention Centre between 1989 and 1997. Syndromic and symptomatic outcome were determined with the Brief Psychiatric Rating Scale; functional outcome with the Quality of Life Scale and Premorbid Adjustment Scale subitems. RESULTS: Symptomatic outcome was assessed in 67 patients at 6 months and 61 patients at 12 months, and functional outcome in 56 patients at 6 months and 49 patients at 12 months. Logistic regressions were conducted on 46 and 43 patients, respectively, to explore predictors of outcome. While 90% of patients achieved syndromic recovery at 6 and 12 months, 40% had not recovered symptomatically at 6 and 12 months, still presenting with anxiety or depression. A total of 66% of patients at 6 months and 61% of patients at 12 months failed to return to previous level of functioning. Age at intake, family history of affective disorder, illicit drug use and functional recovery at 6 months predicted functional outcome at 12 months. CONCLUSIONS: This study confirms poor symptomatic and functional outcome after first-episode psychotic mania. It suggests possible usefulness of early intervention strategies in bipolar disorders and need for developing specific interventions addressing anxiety, depression and substance abuse comorbidity. PMID- 16696824 TI - Treatment adherence with antipsychotic medications in bipolar disorder. AB - OBJECTIVES: Atypical antipsychotic medications are a relatively new, increasingly prominent component of the treatment armamentarium for bipolar disorder. Information on adherence with antipsychotics among individuals with bipolar disorder in general, and atypical antipsychotics in particular, is currently quite limited. Using data from the VA National Psychosis Registry, we examined adherence with antipsychotic medications among patients with bipolar disorder (n = 73,964). METHODS: Antipsychotic medication adherence among veterans with bipolar disorder was evaluated using the medication possession ratio and categorizing patients into three groups: fully adherent, partially adherent and non-adherent. We compared characteristics of bipolar patients who received versus those who did not receive antipsychotic medication, and also identified predictors of poor adherence with antipsychotic medications. RESULTS: Approximately 45% (n = 32,993) of all individuals with bipolar disorder were prescribed antipsychotic medication. Individuals who were prescribed antipsychotic medications were younger and more often had comorbid substance abuse or post-traumatic stress disorder compared to individuals with bipolar disorder who were not prescribed antipsychotic medication. Just over half (51.9%) of individuals appear to be fully adherent with antipsychotic medications, while 48.1% of individuals are either partially adherent or non-adherent with antipsychotic medications. Factors associated with treatment non-adherence were younger age, minority ethnicity, comorbid substance abuse and homelessness. CONCLUSIONS: Treatment non-adherence is a major issue for close to half of individuals with bipolar disorder prescribed antipsychotic medication. Additional studies are needed to better understand treatment adherence within the full range of pharmacologic therapies among individuals with bipolar disorder. PMID- 16696825 TI - Urbanicity during upbringing and bipolar affective disorders in Denmark. AB - BACKGROUND: It has been suggested that known or suspected risk factors for schizophrenia may also be of importance for other psychoses, but the empirical evidence regarding this is limited. Urbanicity of place of birth and during upbringing has been shown to be related to the risk of schizophrenia. Few studies of urbanicity in relation to bipolar affective disorder exist. OBJECTIVE: To investigate the potential association between urbanicity at birth and during upbringing and the risk of bipolar affective disorder. METHOD: Using data from the Danish Civil Registration System, we established a population-based cohort of 2.04 million people born in Denmark during 1956-1986, which included information on place of residence during upbringing. Bipolar affective disorder in cohort members was identified by linkage with the Danish Psychiatric Central Register. RESULTS: Overall, 2232 people developed bipolar affective disorder during 1971 2001. We found evidence of an increased risk associated with residence in the provincial city; individuals, who at the 15th birthday lived in the provincial city, had a risk of 1.23 (1.08-1.41). This increased risk was explained solely by an increased risk associated with residence in Aarhus; at the 15th birthday, people residing in Aarhus - the largest of the three Danish provincial cities - had a 1.83 (1.56-2.14) increased risk of bipolar affective disorder (p < 0.001). Urbanicity during upbringing (p = 0.13) had no significant effects on the risk of bipolar affective disorder. CONCLUSIONS: We found no evidence of a dose-response relationship between urbanicity at birth (and during upbringing) and the risk of bipolar affective disorders in Denmark, but found some evidence that the diagnostic practices used in Aarhus differed from the rest of Denmark. PMID- 16696826 TI - Preliminary evidence of differential relations between prefrontal cortex metabolism and sustained attention in depressed adults with bipolar disorder and healthy controls. AB - OBJECTIVES: To assess the relations between sustained attention as assessed by the Continuous Performance Test (CPT) and subgenual and dorsolateral prefrontal cortex metabolism in depressed patients with bipolar disorders and healthy controls. DESIGN: Cross-sectional case-control design. METHODS: Cerebral metabolic rates were assessed with 18F-fluoro-deoxyglucose and positron emission tomography (PET) in the regions of interest defined on co-registered structural magnetic resonance images in eight medication-free, depressed bipolar disorder patients and 27 healthy control participants. PET scans were obtained in a resting state and the CPT was administered within 1 week of the PET scan. RESULTS: Although there were no statistically significant differences in performance on the CPT or in cerebral metabolism between the two groups, our analyses revealed differential relations between the CPT and metabolism across the groups. Decreased subgenual prefrontal metabolism was associated with slower hit rate reaction time and more omission errors in the bipolar group, but not the control group. Decreased dorsolateral prefrontal metabolism in the bipolar group, but not the control group, was associated with more commission errors. CONCLUSIONS: This study extends previous neuroimaging findings of structural and functional relevance of the prefrontal region with attention to include depressed states in bipolar disorder. The results are consistent with interpretations that decreased prefrontal activity may represent failure to activate some areas of inhibitory control. Decreasing subgenual prefrontal cortex metabolism appears to relate to decreased attention whereas the decreased dorsolateral prefrontal cortex metabolism relates more to decreased inhibitory control. PMID- 16696828 TI - Serum calcium-independent phospholipase A2 activity in bipolar affective disorder. AB - OBJECTIVE: Phospholipases A2 (PLA2) are a family of enzymes involved in membrane phospholipid metabolism and cell signalling. The gene encoding one form, type VI calcium-independent phospholipase A2, is located in a region of DNA that may contain a gene important in the aetiology of psychosis. Moreover, the activity of calcium-independent PLA2 is reported to be elevated in the blood and brain of patients with schizophrenia. In this study we determined whether a similar change takes place in patients with bipolar disorder with and without a history of psychosis. METHODS: Serum calcium-independent and -dependent PLA2 activities were determined in 24 patients with bipolar I disorder. RESULTS: Serum calcium independent and -dependent PLA2 activities in bipolar cases did not differ significantly from that in healthy volunteers (HVs). However, calcium-independent PLA2 activity was significantly (p < 0.05) higher in patients with a history of psychosis compared with those with no history of psychosis (by 55%) or to HVs (by 31%). CONCLUSIONS: Our data suggest that a subset of bipolar I disorder patients with a history of psychosis have elevated calcium-independent PLA2 activity. Given that this enzyme activity is also increased in schizophrenia, elevated rates of phospholipid turnover mediated by the enzyme could represent a common biochemical feature of psychotic illness. PMID- 16696829 TI - The prospective impact of sleep duration on depression and mania. AB - OBJECTIVE: Many patients report sleeping less than 6 h per night during episodes of depression and mania. This type of sleep deficit may also be a risk factor for subsequent mood episodes; however, the long-term impact of sleep deficit remains unclear. The current study is among few longitudinal studies to assess the prospective effect of sleep deficit on depression and mania. METHODS: A subsample of 54 individuals from a longitudinal study of bipolar I disorder was selected. Participants entered the study during a mood episode. Baseline symptom data were collected at month 4 to allow for recovery from the initial episode, sleep was assessed at month 6, and follow-up symptom data were obtained during months 7-12. RESULTS: Sleep deficit predicted depressive symptoms across the 6-month follow-up but not mania. CONCLUSIONS: It is likely that the impact of sleep deficit on mania was probably missed because assessments covered a full month. Monitoring sleep duration may help predict depression in bipolar disorder and provide an opportunity for targeting intervention. PMID- 16696827 TI - Decrease in creatine kinase messenger RNA expression in the hippocampus and dorsolateral prefrontal cortex in bipolar disorder. AB - OBJECTIVES: Bipolar disorder (BPD) affects more than 2 million adults in the USA and ranks among the top 10 causes of worldwide disabilities. Despite its prevalence, very little is known about the etiology of BPD. Recent evidence suggests that cellular energy metabolism is disturbed in BPD. Mitochondrial function is altered, and levels of high-energy phosphates, such as phosphocreatine (PCr), are reduced in the brain. This evidence has led to the hypothesis that deficiencies in energy metabolism could account for some of the pathophysiology observed in BPD. To further explore this hypothesis, we examined levels of creatine kinase (CK) mRNA, the enzyme involved in synthesis and metabolism of PCr, in the hippocampus (HIP) and dorsolateral prefrontal cortex (DLPFC) of control, BPD and schizophrenia subjects. METHODS: Tissue was obtained from the Harvard Brain Tissue Resource Center. Real-time quantitative polymerase chain reaction (HIP, DLPFC) and gene expression microarrays (HIP) were employed to compare the brain and mitochondrial 1 isoforms of CK. RESULTS: Both CK isoforms were downregulated in BPD. Furthermore, mRNA transcripts for oligodendrocyte-specific proteins were downregulated in the DLPFC, whereas the mRNA for the neuron-specific protein microtubule-associated protein 2 was downregulated in the HIP. CONCLUSION: Although some of the downregulation of CK might be explained by cell loss, a more general mechanism seems to be responsible. The downregulation of CK transcripts, if translated into protein levels, could explain the reduction of high-energy phosphates previously observed in BPD. PMID- 16696830 TI - Increased oxidative stress after repeated amphetamine exposure: possible relevance as a model of mania. AB - BACKGROUND: Acute mania can be modeled in animals using D-amphetamine (AMPH). Acute AMPH injections are associated with monoamine depletion, loss of neurofilaments and neurite degeneration. However, the precise mechanisms underlying AMPH-induced neurotoxicity are still unclear. Several studies have demonstrated that oxidative stress may play a role in the behavioral and neurochemical changes observed after AMPH administration. METHODS: The effects of a single and repeated injections (seven daily injections) of AMPH administered intraperitonially on locomotion and the production of lipid and protein oxidative markers in rat cortex, striatum and hippocampus were assessed. Locomotion was assessed in an open-field task and markers of oxidative stress were assessed in brain tissue. RESULTS: Both single and repeated injections of AMPH increased protein carbonyl formation in rat brain. Repeated exposure to AMPH induced an additional increase in thiobarbituric acid reactive species in brain tissue. CONCLUSIONS: Longer periods of exposure to AMPH were associated with increased oxidative stress in rat brain. This adds to the notion that repeated manic episodes may be associated with greater brain damage and, therefore, poorer outcomes. PMID- 16696831 TI - Anterior cingulate subregion volumes and executive function in bipolar disorder. AB - OBJECTIVE: Although research findings suggest a relationship between the function of anterior cingulate cortex (ACC) and both cognitive ability and the pathophysiology of bipolar disorder (BPD), few studies have examined cognitive correlates of specific ACC subregion volumes in BPD. Therefore, the primary aim of this study was to examine the relationship between magnetic resonance imaging (MRI)-derived gray and white matter volumes of ACC subregions (caudal, rostral, and subgenual) and performance on tests of executive function in 27 patients with BPD and 22 healthy subjects. METHODS: 1.5T MRI and neuropsychological assessment were conducted with all participants. RESULTS: MANCOVA revealed statistically significant group differences in performance on executive function measures. However, no group differences were observed in any of the ACC white matter or gray matter regions of interest. Multiple regression analyses revealed that rostral and subgenual gray matter each interacted significantly with group in predicting performance on the Wisconsin Card Sorting Test. In addition, a significant interaction was observed between group and both rostral gray and white matter in predicting performance on the Trail Making Test. CONCLUSIONS: The results of this preliminary study support the extant literature that suggests that patients with BPD perform more poorly than healthy subjects on tests of executive function. Furthermore, the relationship between ACC subregion volumes and cognitive test performance was found to differ between patients with BPD and healthy subjects, despite comparable ACC volumes in the two groups. PMID- 16696832 TI - Effects of lamotrigine in patients with bipolar disorder and alcohol dependence. AB - BACKGROUND: Bipolar disorder is significantly associated with alcohol use disorders. Anticonvulsant drugs are used in the treatment of bipolar disorder and they have also been used to treat alcohol dependence. The purpose of the present study was to evaluate tolerance and safety of lamotrigine in a dual-diagnosis population presenting bipolar disorder and alcohol dependence. Open-label lamotrigine was examined in 28 outpatients with DSM-IV bipolar disorder and alcohol dependence. Lamotrigine was added to existing medication regimens. METHOD: Lamotrigine was started at a dose of 25 mg/day and titrated to a maximum dose of 300 mg/day. Subjects received a baseline evaluation which included a Structured Clinical Interview for DSM-IV (SCID) and weekly assessments for 12 weeks with the Hamilton Rating Scale for Depression (HAM-D), Young Mania Rating Scale (YMRS), Brief Psychiatric Rating Scale (BPRS), Severity of Alcohol Dependence Scale (SADS), a Visual Analogue Scale for Craving severity (VASC), and alcohol consumption. The concentration of carbohydrate-deficient transferrin (CDT) was used as an indirect measure of alcohol consumption. The sample consisted of 18 men and 10 women diagnosed with alcohol dependence and bipolar disorder I (n = 21) or bipolar disorder II (n = 7), with a mean age of 36.5 +/- 7.7 years. RESULTS: Significant improvement was observed in HAM-D, YMRS, and BPRS scores (p < 0.01). Craving and CDT also significantly decreased (p < 0.001). Lamotrigine was well tolerated with no dropout subjects due to adverse events. CONCLUSION: Lamotrigine is safe and well tolerated in this sample and associated with improvement in mood, alcohol craving and alcohol consumption. A placebo controlled study would be of interest. PMID- 16696833 TI - Duplication 8q22.1-q24.1 associated with bipolar disorder and speech delay. AB - OBJECTIVE: To report a case of a child with bipolar disorder found to have an unbalanced translocation involving the long arm of chromosome 8, a region that has been previously implicated in genome-wide linkage scans. CASE REPORT: A 7 year-old boy with a complex psychiatric symptom presentation including attention deficits, distractibility, impulsivity, pressured speech, sleep disturbance, aggressive behavior, and hypersexuality diagnosed with bipolar disorder. He also showed evidence of borderline intellectual and adaptive functioning and had mild dysmorphic features with a duplication of distal 8q that arose as an unbalanced chromosomal translocation due to a maternal 15p;8q insertion. CONCLUSION: This finding of an unbalanced translocation provides further evidence to support previous linkage studies of a potential causative gene on 8q for bipolar disorder. PMID- 16696834 TI - Aripiprazole use in a pregnant schizoaffective woman. AB - BACKGROUND: There are few or no clinical data on the safety of aripiprazole during pregnancy and lactation. CASE REPORT: A 27-year-old schizoaffective woman conceived while receiving aripiprazole 15 mg/day. The drug was withdrawn during the eighth week of gestation. She relapsed at week 20, and resumed treatment with the drug at a dose of 10 mg/day. She responded well, and the drug was continued through the rest of the pregnancy. The pregnancy was uneventful, but unexplained fetal tachycardia necessitated a cesarean section at full term. The newborn infant was normal in all regards and developed normally during a 6-month follow up. The infant required to be bottle-fed because of a failure of lactation. CONCLUSION: In this case, exposure to aripiprazole during different trimesters of pregnancy was not associated with intrauterine death, morphological teratogenicity, impaired fetal growth, neonatal toxicity, or neurobehavioral teratogenicity. Maternal health was not adversely affected during pregnancy. However, lactation failure occurred after delivery. PMID- 16696835 TI - Maintenance electroconvulsive therapy for recurrent refractory mania. AB - BACKGROUND: Although there are some reports on the efficacy of maintenance electroconvulsive therapy (mECT) in rapid cycling bipolar disorder there is scant data about the use of mECT in the non-rapid cycling form of this condition. PATIENT AND METHODS: We report the case of a 45-year-old man diagnosed with bipolar affective disorder at the age of 32 who had 5 manic episodes in 6 years. These episodes were severe, required hospitalization and had poor response to lithium carbonate, sodium valproate, carbamazepine, haloperidol and chlorpromazine (either alone or in combination). The patient was treated with electroconvulsive therapy (ECT) during his last hospitalization and since then his treatment has included mECT and chlorpromazine (400 mg/day) and he has remained euthymic during the past 21 months. CONCLUSION: This case describes the utility of mECT in medication refractory bipolar disorder. PMID- 16696836 TI - Electroconvulsive therapy: an effective therapy of medication-resistant bipolar disorder. AB - OBJECTIVES: We report our experience with a medication-refractory patient with bipolar I disorder, who remained clinically stable under continuation electroconvulsive therapy (C-ECT). We found ECT to be effective in providing functionality for a patient when various medications had failed. METHODS AND RESULTS: A 53-year-old woman with a 31-year history of recurrent manic and depressive episodes poorly responsive to medications warranted an ECT trial. Remission was achieved after 12 treatments and C-ECT at weekly intervals was used as maintenance therapy. C-ECT was continued for 37 months and 85 treatments, and discontinued when anesthetic difficulties precluded further treatment. Three months after the last ECT, she was readmitted in a severe manic relapse. Restarting ECT caused rapid remission. There are no signs of cognitive deterioration during C-ECT. CONCLUSION: Long-term C-ECT is an effective and safe prophylactic treatment in individual treatment-resistant patients with bipolar disorder. PMID- 16696838 TI - Abstracts of the XXVIIth International Congress of the World Federation of Hemophilia, Vancouver, Canada, 21-25 May, 2006. PMID- 16696837 TI - ECT in therapy-resistant mania: does it have a place? PMID- 16696839 TI - Dichotic listening with forced attention in patients with temporal lobe epilepsy: significance of left hemisphere cognitive dysfunction. AB - Fifty right-handed patients with focal temporal lobe epilepsy were administered a dichotic listening test with consonant-vowel syllables under non-forced, forced right and forced left attention conditions, and a neuropsychological test battery. Dichotic listening performance was compared in subgroups with and without left hemisphere cognitive dysfunction, measured by the test battery, and in subgroups with left and right temporal epileptic focus. Left hemisphere cognitive dysfunction led to more correct responses to left ear stimuli in all three attention conditions, and fewer correct responses to right ear stimuli in the non-forced attention condition. This was probably caused by basic left hemisphere perceptual dysfunction. Dichotic listening was less affected by a left sided epileptic focus than by left hemisphere cognitive dysfunction. General cognitive functioning influenced dichotic listening performance stronger in forced than in non-forced attention conditions. Larger cerebral networks were probably involved in the forced attention conditions due to the emphasis on conscious effort. PMID- 16696840 TI - Orientation dependent mental representations following real-world navigation. AB - The purpose of this study was to examine whether the mental representations acquired from real-world navigation are encoded in a single, specific orientation. Previous research has revealed an inconsistent pattern of results. In the present study, participants explored a university campus. In two reaction time tasks, spatial judgments were then made relative to four imagined headings within the explored environment. The main result was that one of the headings was encoded in a distinct manner: When making left-right judgments performance on this heading was generally superior by comparison with the other orientations, and when making front-back judgments the same heading was the only one in which an advantage of front over back responses was observed. The finding that the headings were not encoded equally indicates that real-world navigation can lead to orientation specific representations. PMID- 16696841 TI - The face you recognize may not be the one you saw: memory conjunction errors in individuals with or without learning disability. AB - Memory conjunction errors, that is, when a combination of two previously presented stimuli is erroneously recognized as previously having been seen, were investigated in a face recognition task with drawings and photographs in 23 individuals with learning disability, and 18 chronologically age-matched controls without learning disability. Compared to the controls, individuals with learning disability committed significantly more conjunction errors, feature errors (one old and one new component), but had lower correct recognition, when the results were adjusted for different guessing levels. A dual-processing approach gained more support than a binding approach. However, neither of the approaches could explain all of the results. The results of the learning disability group were only partly related to non-verbal intelligence. PMID- 16696842 TI - A Swedish non-word repetition test for preschool children. AB - A Swedish non-word repetition test for preschool children was constructed with the aim of measuring short-term phonological memory, which has been shown to correlate with language development and language proficiency. Normative data based on results for 200 children four to six years of age are presented. An item analysis of the original 30-item test resulted in a shortened 24 non-word version found suitable for four- to six-year old children. Results for this shortened version, which had a Cronbach's alpha of 0.73, showed longer non-words to be more difficult to repeat than shorter ones, and significant effects of age and compatibility of the tester's and the child's accent, but not of gender. The results are discussed within the framework of Baddeley's working memory model. PMID- 16696843 TI - The ethic of care development: a longitudinal study of moral reasoning among practical-nursing, social-work and law-enforcement students. AB - The study investigated changes in care-based moral reasoning, in the context of justice development over the 2-year period among practical-nursing, bachelor degree social-work and law-enforcement students (N = 59). Main measures were Skoe's Ethic of Care Interview and Colby et al.'s Moral Judgment Interview. Of the participants 34% progressed in care reasoning, and 48% in justice reasoning. Social-work and nursing students progressed in care reasoning, and all groups progressed in justice reasoning. One participant (1.7%) regressed in care reasoning. Care and justice reasoning were parallel in terms of internal consistency, and they were positively related to each other. Findings suggest that care reasoning follows a developmental sequence, involving three main and two transitional levels, as suggested by Gilligan (1982). Main levels include self-concern (Level 1), caring for others (Level 2), and balanced caring for self and others (Level 3). PMID- 16696844 TI - Social support, locus of control, and depressive symptoms in hemodialysis patients. AB - This study aimed to reveal how social support and psychological well-being association might differ with locus of control orientation for chronically ill patients receiving a restrictive and unpleasant medical treatment (i.e., hemodialysis). Data were collected from 104 hemodialysis patients. After controlling for the variance accounted for by gender and duration of dialysis, for patients with internal locus of control, lack of "perceived social support" was found to be associated with depressive symptoms. On the other hand, for patients with external locus of control, the same analysis revealed that lack of "satisfaction from the received social support" was associated with depressive symptoms. Thus, for hemodialysis patients the variables associated with depressive symptoms varied with their locus of control orientations. Furthermore, the present study underlined the importance of considering different aspects of social support while studying with chronically ill patients. PMID- 16696845 TI - Moving towards a favorable image: the self-presentational benefits of exercise and physical activity. AB - The purpose of this study was to investigate the exercise stereotype phenomenon and the moderating effects of exerciser impression motivation and construction on this stereotype in a sample of 176 female and 96 male Swedish university students. The participants read a description of one of the following female targets: a typical exerciser, an active living target, an excessive exerciser, a non-exerciser, or a control target, and then rated these targets on 12 personality (e.g., lazy-hard worker, dependent-independent) and 8 physical (e.g., scrawny-muscular, sick-healthy) dimensions. They also completed the Self Presentation in Exercise Questionnaire, measuring motivation to self-present as an exerciser and the propensity to construct the image of an exerciser. MANCOVAs revealed a significant main effect for both personality and physical attributes (p < 0.05). In general, the typical exerciser and active living targets received the most favorable ratings, especially on the physical attributes, whereas the excessive exerciser obtained the least positive ratings. Exerciser impression motivation moderated the exercise status/rating relationship for the physical attributes only. Differences between Swedish and North American students' impressions of exercisers and non-exercisers are discussed. PMID- 16696846 TI - Personality traits in women with Turner syndrome. AB - Turner syndrome is caused by the total or partial absence of, or structural abnormalities in, one sex chromosome. A personality style characterized by traits related to low neuroticism and high extroversion has been reported for females with Turner syndrome, indicating possible genetic factors related to personality development. This study examines personality traits in 37 Swedish women with Turner syndrome using the Karolinska Scales of Personality Inventory. Compared to reference data, the subjects rated themselves as more sensation seeking, less detached, less verbally aggressive and less irritated. These results partially support previous findings. The subjects were divided into three karyotype groups: 45,X (n = 13), 45,X/46,XX or 45,X/45,XY (n = 13), and structural abnormalities (n = 11). Contrary to some previous findings, no differences in personality were found between the karyotype groups. PMID- 16696847 TI - Does neuroscience hold promise for the further development of behavior therapy? The case of emotional change after exposure in anxiety and depression. AB - Over the last few years, the integration of biological and psychological models has become increasingly important in clinical psychology. This paper presents a neurobiological framework for exploring how neuroscience research may be valuable in the context of new developments in behavior therapy. The importance of the cortical-subcortical circuitry of emotion processing for investigating the working mechanisms of exposure therapy is illustrated by current neuroimaging research. The implication is that during exposure therapy the focus of attention must be fully directed towards the emotional content of the threatening situation to facilitate prefrontal control over the amygdala and that new contextual information, of emotional salience, must be added to create inhibitory projections from the hippocampus. In addition, some possible mechanisms of action for different therapeutic approaches to both anxiety and depression and the role of neuroimaging techniques in predicting treatment outcome are discussed. PMID- 16696848 TI - Sustained elevation of extracellular adenosine and activation of A1 receptors underlie the post-ischaemic inhibition of neuronal function in rat hippocampus in vitro. AB - Adenosine is released from the compromised brain and exerts a predominately neuroprotective influence. However, the time-course of adenosine release and its relationship to synaptic activity during metabolic stress is not fully understood. Here, we describe experiments using an enzyme-based adenosine sensor to show that adenosine potently (IC50 approximately 1 microm) inhibits excitatory synaptic transmission in area CA1 during oxygen/glucose deprivation ('ischaemia'), and that the prolonged post-ischaemic presence of extracellular adenosine sustains the depression of the field excitatory postsynaptic potential (fEPSP). N-methyl-D-aspartate (NMDA) receptor antagonism promotes post-ischaemic recovery of the fEPSP, in parallel with reduced release of adenosine. Paradoxically, however, after ischaemia the fEPSP recovers in the face of concentrations of adenosine capable of fully eliminating synaptic transmission during ischaemia. This hysteresis is not prevented by NMDA receptor antagonism, is observed during repeated ischaemia when adenosine release is reduced, and does not reflect desensitization of adenosine A1 receptors. We conclude that adenosine exerts powerful inhibitory actions on excitatory synaptic transmission both during, and for some considerable time after, ischaemia. Therapeutic strategies designed to exploit both the continued presence of adenosine and activity of A1 receptors could provide benefits in individuals who have suffered acute injury to the CNS. PMID- 16696849 TI - Chronic pain-induced emotional dysfunction is associated with astrogliosis due to cortical delta-opioid receptor dysfunction. AB - It has been widely recognized that chronic pain could cause physiological changes at supraspinal levels. The delta-opioidergic system is involved in antinociception, emotionality, immune response and neuron-glia communication. In this study, we show that mice with chronic pain exhibit anxiety-like behavior and an increase of astrocytes in the cingulate cortex due to the dysfunction of cortical delta-opioid receptor systems. Using neural stem cells cultured from the mouse embryonic forebrain, astrocyte differentiation was clearly observed following long-term exposure to the selective delta-opioid receptor antagonist, naltrindole. We also found that micro-injection of either activated astrocyte or astrocyte-conditioned medium into the cingulate cortex of mice aggravated the expression of anxiety-like behavior. Our results indicate that the chronic pain process promotes astrogliosis in the cingulate cortex through the dysfunction of cortical delta-opioid receptors. This phenomenon may lead to emotional disorders including aggravated anxiety under chronic pain-like state. PMID- 16696850 TI - Immunocytochemical localization of 3-methylcrotonyl-CoA carboxylase in cultured ependymal, microglial and oligodendroglial cells. AB - To evaluate the ability of ependymal, microglial and oligodendroglial cells to degrade leucine, the presence of 3-methylcrotonyl-CoA carboxylase (MCC) was investigated in cultures of these cells. MCC is a biotin-containing heterodimeric enzyme that is specific for the irreversible part of the leucine catabolic pathway. It has been reported previously that in cell culture MCC is expressed in astrocytes and a subpopulation of neurones. In the present study ependymal, microglial and oligodendroglial cell cultures, derived from the brains of newborn rats, were examined for the expression of MCC by RT-PCR, western blotting and immunocytochemistry. The results of RT-PCR and western blotting showed the presence of mRNA as well as protein of both subunits of MCC in ependymal, microglial and oligodendroglial cell cultures. Immunocytochemical investigation of the cellular and subcellular distribution of MCC demonstrated a mitochondrial location of MCC in all neuroglial cell types investigated. The ubiquitous expression of MCC in glial cells demonstrates the ability of the cells to engage in the catabolism of leucine transported into the brain, mainly for the generation of energy. PMID- 16696851 TI - Substance P release evoked by capsaicin or potassium from rat cultured dorsal root ganglion neurons is conversely modulated with bradykinin. AB - To clarify the molecular mechanism of substance P (SP) release from dorsal root ganglion (DRG) neurons, we investigated the involvement of several intracellular effectors in the regulation of SP release evoked by capsaicin, potassium or/and bradykinin. Bradykinin-evoked SP release from cultured adult rat DRG neurons was attenuated by either the mitogen-activated protein kinase kinase (MEK) inhibitor (U0126) or cycloheximide. As the long-term exposure of DRG neurons to bradykinin (3 h) resulted in extracellular signal-regulated kinase (ERK) phosphorylation at an early stage and thereafter induced cyclooxygenase-2 (COX-2) protein expression, which both contribute to the SP release triggered by bradykinin B2 receptor. The long-term exposure of DRG neurons to bradykinin enhanced the SP release by capsaicin, but attenuated that by potassium. Interestingly, the inositol 1,4,5-triphosphate (IP3)-induced calcium release blocker [2-aminoethyl diphenylborinate (2-APB)] not only inhibited the potassium-evoked SP release, but also completely abolished the enhancement of capsaicin-induced SP release by bradykinin from cultured DRG neurons. Together, these findings suggest that the molecular mechanisms of SP release by bradykinin involve the activation of MEK, and also require the de novo protein synthesis of COX-2 in DRG neurons. The IP3 dependent calcium release could be involved in the processes of the regulation by bradykinin of capsaicin-triggered SP release. PMID- 16696852 TI - Identification of phenylethanolamine N-methyltransferase gene expression in stellate ganglia and its modulation by stress. AB - Phenylethanolamine N-methyltransferase (PNMT, EC 2.1.1.28) is the terminal enzyme of the catecholaminergic pathway converting noradrenaline to adrenaline. Although preferentially localized in adrenal medulla, evidence exists that PNMT activity and gene expression are also present in the rat heart, kidney, spleen, lung, skeletal muscle, thymus, retina and different parts of the brain. However, data concerning PNMT gene expression in sympathetic ganglia are still missing. In this study, our effort was focused on identification of PNMT mRNA and/or protein in stellate ganglia and, if present, testing the effect of stress on PNMT mRNA and protein levels in this type of ganglia. We identified both PNMT mRNA and protein in stellate ganglia of rats and mice, although in much smaller amounts compared with adrenal medulla. PNMT gene expression and protein levels were also increased after repeated stress exposure in stellate ganglia of rats and wild-type mice. Similarly to adrenal medulla, the immobilization-induced increase was probably regulated by glucocorticoids, as determined indirectly using corticotropin releasing hormone knockout mice, where immobilization-induced increase of PNMT mRNA was suppressed. Thus, glucocorticoids might play an important role in regulation of PNMT gene expression in stellate ganglia under stress conditions. PMID- 16696853 TI - A yeast 2-hybrid analysis of human GTP cyclohydrolase I protein interactions. AB - The yeast 2-hybrid system was used to identify protein domains involved in the oligomerization of human guanosine 5'-triphosphate (GTP) Cyclohydrolase I (GCH1) and the interaction of GCH1 with its regulatory partner, GCH1 feedback regulatory protein (GFRP). When interpreted within the structural framework derived from crystallography, our results indicate that the GCH1 N-terminal alpha-helices are not the only domains involved in the formation of dimers from monomers and also suggest an important role for the C-terminal alpha-helix in the assembly of dimers to form decamers. Moreover, a previously unknown role of the extended N terminal alpha-helix in the interaction of GCH1 and GFRP was revealed. To discover novel GCH1 protein binding partners, we used the yeast 2-hybrid system to screen a human brain library with GCH1 N-terminal amino acids 1-96 as prey. This protruding extension of GCH1 contains two canonical Type-I Src homology-3 (SH3) ligand domains located within amino acids 1-42. Our screen yielded seven unique clones that were subsequently shown to require amino acids 1-42 for binding to GCH1. The interaction of one of these clones, Activator of Heat Shock 90 kDa Protein (Aha1), with GCH1 was validated by glutathione-s-transferase (GST) pull-down assay. Although the physiological relevance of the Aha1-GCH1 interaction requires further study, Aha1 may recruit GCH1 into the endothelial nitric oxide synthase/heat shock protein (eNOS/Hsp90) complex to support changes in endothelial nitric oxide production through the local synthesis of BH4. PMID- 16696854 TI - Molecular morphology and toxicity of cytoplasmic prion protein aggregates in neuronal and non-neuronal cells. AB - Recent studies have revealed that accumulation of prion protein (PrP) in the cytoplasm results in the production of aggregates that are insoluble in non-ionic detergents and partially resistant to proteinase K. Transgenic mice expressing PrP in the cytoplasm develop severe ataxia with cerebellar degeneration and gliosis, suggesting that cytoplasmic PrP may play a role in the pathogenesis of prion diseases. The mechanism of cytoplasmic PrP neurotoxicity is not known. In this report, we determined the molecular morphology of cytoplasmic PrP aggregates by immunofluorescence and electron microscopy, in neuronal and non-neuronal cells. Transient expression of cytoplasmic PrP produced juxtanuclear aggregates reminiscent of aggresomes in human embryonic kidney 293 cells, human neuroblastoma BE2-M17 cells and mouse neuroblastoma N2a cells. Time course studies revealed that discrete aggregates form first throughout the cytoplasm, and then coalesce to form an aggresome. Aggresomes containing cytoplasmic PrP were 1-5-microm inclusion bodies and were filled with electron-dense particles. Cytoplasmic PrP aggregates induced mitochondrial clustering, reorganization of intermediate filaments, prevented the secretion of wild-type PrP molecules and diverted these molecules to the cytoplasm. Cytoplasmic PrP decreased the viability of neuronal and non-neuronal cells. We conclude that any event leading to accumulation of PrP in the cytoplasm is likely to result in cell death. PMID- 16696856 TI - Role of delta-opioid receptor function in neurogenesis and neuroprotection. AB - The present study was undertaken to evaluate the implication of delta-opioid receptor function in neurogenesis and neuroprotection. We found that the stimulation of delta-opioid receptors by the selective delta-opioid receptor agonist SNC80 [(+)-4-[(alphaR)-alpha-((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl) 3-methoxybenzyl]-N,N-diethylbenzamide] (10 nm) promoted neural differentiation from multipotent neural stem cells obtained from embryonic C3H mouse forebrains. In contrast, either a selective micro-opioid receptor agonist, [D-Ala2, N-Me Phe4, Gly5-ol]-enkephalin (DAMGO), or a specific kappa-opioid receptor agonist, ( )-trans-(1S,2S)-U-50488 hydrochloride (U50,488H), had no such effect. In addition to neural differentiation, the increase in cleaved caspase 3-like immunoreactivity induced by H2O2 (3 microm) was suppressed by treatment with SNC80 in cortical neuron/glia co-cultures. These effects of SNC80 were abolished by a Trk-dependent tyrosine kinase inhibitor: (8R*,9S*,11S*)-(-)-9-hydroxy-9 methoxycarbonyl-8-methyl-2,3,9,10-tetrahydro-8,11-epoxy-1H,8H,11H-2,7b,11a triazadibenzo(a,g)cycloocta(cde)trinden-1-one (K-252a). The SNC80-induced neural differentiation was also inhibited by treatment with the protein kinase C (PKC) inhibitor, phosphatidylinositol 3-kinase (PI3K) inhibitor, mitogen-activated protein kinase kinase (MEK) inhibitor or Ca2+/calmodulin-dependent protein kinase II (CaMKII) inhibitor. These findings raise the possibility that delta-opioid receptors play a crucial role in neurogenesis and neuroprotection, mainly through the activation of Trk-dependent tyrosine kinase, which could be linked to PI3K, PKC, CaMKII and MEK. PMID- 16696855 TI - Genetic selection of sox1GFP-expressing neural precursors removes residual tumorigenic pluripotent stem cells and attenuates tumor formation after transplantation. AB - Because of their ability to proliferate and to differentiate into diverse cell types, embryonic stem (ES) cells are a potential source of cells for transplantation therapy of various diseases, including Parkinson's disease. A critical issue for this potential therapy is the elimination of undifferentiated cells that, even in low numbers, could result in teratoma formation in the host brain. We hypothesize that an efficient solution would consist of purifying the desired cell types, such as neural precursors, prior to transplantation. To test this hypothesis, we differentiated sox1-green fluorescent protein (GFP) knock-in ES cells in vitro, purified neural precursor cells by fluorescence-activated cell sorting (FACS), and characterized the purified cells in vitro as well as in vivo. Immunocytofluorescence and RT-PCR analyses showed that this genetic purification procedure efficiently removed undifferentiated pluripotent stem cells. Furthermore, when differentiated into mature neurons in vitro, the purified GFP+ cell population generated enriched neuronal populations, whereas the GFP- population generated much fewer neurons. When treated with dopaminergic inducing signals such as sonic hedgehog (SHH) and fibroblast growth factor-8 (FGF8), FACS purified neural precursor cells responded to these molecules and generated dopaminergic neurons as well as other neural subtypes. When transplanted, the GFP+ cell population generated well contained grafts containing dopaminergic neurons, whereas the GFP- population generated significantly larger grafts (about 20-fold) and frequent tumor-related deaths in the transplanted animals. Taken together, our results demonstrate that genetic purification of neural precursor cells using FACS isolation can effectively remove unwanted proliferating cell types and avoid tumor formation after transplantation. PMID- 16696857 TI - Clonal analysis of palmar fibromatosis: a study whether palmar fibromatosis is a real tumor. AB - BACKGROUND: Palmar fibromatosis that arises in the palmar soft tissue is characterized by infiltrative growth with a tendency toward local recurrence but does not metastasize. This study investigated the clonality of this process in twelve female patients, each with a single lesion, by examining the pattern of X chromosome inactivation. METHODS: Hematoxylin and eosin stained sections of formalin-fixed, paraffin-embedded tissues were microdissected by laser capture microdissection to obtain the proliferative spindle cells. Tumor cells were isolated from the sections of rectum adenocarcinoma, and used for positive control. The genomic DNAs was extracted with phenol-chloroform, digested with a methylation-sensitive restriction endonuclease HpaII, and amplified by polymerase chain reaction (PCR), using primers targeted to a highly polymorphic short tandem repeat (STR) of the human androgen receptor gene (HUMARA). RESULTS: Among the twelve samples, three samples failed amplification, one sample showed homozygosity which was not suitable for further analysis, eight samples were successfully amplified, and showed a random X chromosome inactivation pattern, suggesting polyclonality of these lesions. CONCLUSION: The current findings suggest that palmar fibromatosis is a reactive proliferation rather than a clonal neoplasm. PMID- 16696858 TI - Gender differentials in the evolution of cigarette smoking habits in a general European adult population from 1993-2003. AB - BACKGROUND: Describe the recent evolution of cigarette smoking habits by gender in Geneva, where incidence rates of lung cancer have been declining in men but increasing in women. METHODS: Continuous cross-sectional surveillance of the general adult (35-74 yrs) population of Geneva, Switzerland for 11 years (1993 2003) using a locally-validated smoking questionnaire, yielding a representative random sample of 12,271 individuals (6,164 men, 6,107 women). RESULTS: In both genders, prevalence of current cigarette smoking was stable over the 11-year period, at about one third of men and one quarter of women, even though smoking began at an earlier age in more recent years. Older men were more likely to be former smokers than older women. Younger men, but not women, tended to quit smoking at an earlier age. CONCLUSION: This continuous (1993-2003) risk factor surveillance system, unique in Europe, shows stable prevalence of smoking in both genders. However, sharp contrasts in age-specific prevalence of never and former smoking and of ages at smoking initiation indicate that smoking continues a long term decline in men but has still not reached its peak in women. PMID- 16696859 TI - Improvement in quality of life measures in patients with refractory hepatitis C, responding to re-treatment with Pegylated interferon alpha -2b and ribavirin. AB - BACKGROUND: In this paper, we report the health related quality of life (HRQOL) data from patients with hepatitis C viral infection (HCV) who were refractory to prior therapy and had re-treatment with a combination of Pegylated interferon alpha-2b and ribavirin. We hypothesized that the HRQOL will improve in those patients who attain sustained viral response similar to naive patients undergoing treatment for HCV. METHODS: HRQOL data was obtained from 152 patients enrolled into a randomized study for re-treatment of HCV refractory to prior therapy with interferon alpha-2b in combination with ribavirin. The treatment protocol was for 48 weeks and had a high and low dose arm. The HRQOL data was collected at baseline, weeks 24 and 48 of treatment, and at 24 week follow-up after treatment. A repeated measures statistical model was used for comparing the HRQOL domain scores between the responders and non-responders and the treatment groups. The responders and non-responders were also compared to the age and sex adjusted national mean scores. RESULTS: Twenty-five of the 152 (17%) patients achieved a sustained viral response. At baseline, HRQOL is lower in HCV patients compared to national norms. The norm based HRQOL domain scores for the different domains of the SF-36 instrument were as follows: physical functioning = 47.13, role-physical = 46.87, bodily pain = 48.00, general health = 44.01, vitality = 45.39, social functioning = 47.05, role-emotional = 48.88, mental health = 48.76, physical component score 43.26 and mental component score = 46.17. The scores decreased during therapy in those who would be responders and non-responders, but the pattern of change was different. During the treatment, the HRQOL domain scores of responders decrease notably in the domain of vitality. At week 48 vitality scores were worst in responders. 5 of the 8 domain scores were lower compared to baseline in non-responders. At 24 weeks post treatment follow up, HRQOL in those refractory patients who respond to re-treatment tended to be better than the national average in the domains of vitality (p = .06), social functioning (p = .06) and role-emotional (p = .03) while the non-responders improved their scores in domains of physical function and bodily pain. CONCLUSION: We conclude that patients who are to be responders and non-responders behave differently in terms of the HRQOL domain scores when re-treated with a combination of interferon alpha 2b and ribavirin. The responders sustained a significant decrease in the domain score of vitality while 5 of the 8 domain scores decrease in non-responders at the end of treatment. At the end of follow up, in responders, the HRQOL score tended to be better than the national average notably in the domains of role emotional, vitality and social functioning. On the other hand, in non-responders, the domain scores of physical function improve, while that of role-emotional worsened. PMID- 16696860 TI - Gene expression profiling of microglia infected by a highly neurovirulent murine leukemia virus: implications for neuropathogenesis. AB - BACKGROUND: Certain murine leukemia viruses (MLVs) are capable of inducing progressive spongiform motor neuron disease in susceptible mice upon infection of the central nervous system (CNS). The major CNS parenchymal target of these neurovirulent retroviruses (NVs) are the microglia, whose infection is largely coincident with neuropathological changes. Despite this close association, the role of microglial infection in disease induction is still unknown. In this paper, we investigate the interaction of the highly virulent MLV, FrCasE, with microglia ex vivo to evaluate whether infection induces specific changes that could account for neurodegeneration. Specifically, we compared microglia infected with FrCasE, a related non-neurovirulent virus (NN) F43/Fr57E, or mock-infected, both at a basic virological level, and at the level of cellular gene expression using quantitative real time RT-PCR (qRT-PCR) and Afffymetrix 430A mouse gene chips. RESULTS: Basic virological comparison of NN, NV, and mock-infected microglia in culture did not reveal differences in virus expression that provided insight into neuropathogenesis. Therefore, microglial analysis was extended to ER stress gene induction based on previous experiments demonstrating ER stress induction in NV-infected mouse brains and cultured fibroblasts. Analysis of message levels for the ER stress genes BiP (grp78), CHOP (Gadd153), calreticulin, and grp58 in cultured microglia, and BiP and CHOP in microglia enriched fractions from infected mouse brains, indicated that FrCasE infection did not induce these ER stress genes either in vitro or in vivo. To broadly identify physiological changes resulting from NV infection of microglia in vitro, we undertook a gene array screen of more than 14,000 well-characterized murine genes and expressed sequence tags (ESTs). This analysis revealed only a small set of gene expression changes between infected and uninfected cells (<18). Remarkably, gene array comparison of NN- and NV-infected microglia revealed only 3 apparent gene expression differences. Validation experiments for these genes by Taqman real time RT-PCR indicated that only single Ig IL-1 receptor related protein (SIGIRR) transcript was consistently altered in culture; however, SIGIRR changes were not observed in enriched microglial fractions from infected brains. CONCLUSION: The results from this study indicate that infection of microglia by the highly neurovirulent virus, FrCasE, does not induce overt physiological changes in this cell type when assessed ex vivo. In particular, NV does not induce microglial ER stress and thus, FrCasE-associated CNS ER stress likely results from NV interactions with another cell type or from neurodegeneration directly. The lack of NV-induced microglial gene expression changes suggests that FrCasE either affects properties unique to microglia in situ, alters the expression of microglial genes not represented in this survey, or affects microglial cellular processes at a post-transcriptional level. Alternatively, NV-infected microglia may simply serve as an unaffected conduit for persistent dissemination of virus to other neural cells where they produce acute neuropathogenic effects. PMID- 16696861 TI - Cost-effectiveness of microendoscopic discectomy versus conventional open discectomy in the treatment of lumbar disc herniation: a prospective randomised controlled trial [ISRCTN51857546]. AB - BACKGROUND: Open discectomy is the standard surgical procedure in the treatment of patients with long-lasting sciatica caused by lumbar disc herniation. Minimally invasive approaches such as microendoscopic discectomy have gained attention in recent years. Reduced tissue trauma allows early ambulation, short hospital stay and quick resumption of daily activities. A comparative cost effectiveness study has not been performed yet. We present the design of a randomised controlled trial on cost-effectiveness of microendoscopic discectomy versus conventional open discectomy in patients with lumbar disc herniation. METHODS/DESIGN: Patients (age 18-70 years) presenting with sciatica due to lumbar disc herniation lasting more than 6-8 weeks are included. Patients with disc herniation larger than 1/3 of the spinal canal diameter, or disc herniation less than 1/3 of the spinal canal diameter with concomitant lateral recess stenosis or sequestration, are eliglible for participation. Randomisation into microendoscopic discectomy or conventional unilateral transflaval discectomy will take place in the operating room after induction of anesthesia. The length of skin incision is equal in both groups. The primary outcome measure is the functional assessment of the patient, measured by the Roland Disability Questionnaire for Sciatica, at 8 weeks and 1 year after surgery. We will also evaluate several other outcome parameters, including perceived recovery, leg and back pain, incidence of re-operations, complications, serum creatine kinase, quality of life, medical consumption, absenteeism and costs. The study is a randomised prospective multi-institutional trial, in which two surgical techniques are compared in a parallel group design. Patients and research nurses are kept blinded of the allocated treatment during the follow-up period of 2 years. DISCUSSION: Currently, open discectomy is the golden standard in the surgical treatment of lumbar disc herniation. Whether microendoscopic discectomy is more cost-effective than unilateral transflaval discectomy has to be determined by this trial. PMID- 16696862 TI - Exogenous sphingomyelinase increases collagen and sulphated glycosaminoglycan production by primary articular chondrocytes: an in vitro study. AB - We previously established a role for the second messenger ceramide in protein kinase R (PKR)-mediated articular cartilage degradation. Ceramide is known to play a dual role in collagen gene regulation, with the effect of ceramide on collagen promoter activity being dependent on its concentration. Treatment of cells with low doses of sphingomyelinase produces small increases in endogenous ceramide. We investigated whether ceramide influences articular chondrocyte matrix homeostasis and, if so, the role of PKR in this process. Bovine articular chondrocytes were stimulated for 7 days with sphingomyelinase to increase endogenous levels of ceramide. To inhibit PKR, 2-aminopurine was added to duplicate cultures. De novo sulphated glycosaminoglycan and collagen synthesis were measured by adding [35S]-sulphate and [3H]-proline to the media, respectively. Chondrocyte phenotype was investigated using RT-PCR and Western blot analysis. Over 7 days, sphingomyelinase increased the release of newly synthesized sulphated glycosaminoglycan and collagen into the media, whereas inhibition of PKR in sphingomyelinase-treated cells reduced the level of newly synthesized sulphated glycosaminoglycan and collagen. Sphingomyelinase treated chondrocytes expressed col2a1 mRNA, which is indicative of a normal chondrocyte phenotype; however, a significant reduction in type II collagen protein was detected. Therefore, small increments in endogenous ceramide in chondrocytes appear to push the homeostatic balance toward extracellular matrix synthesis but at the expense of the chondrocytic phenotype, which was, in part, mediated by PKR. PMID- 16696863 TI - Failure of non-invasive ventilation in patients with acute lung injury: observational cohort study. AB - INTRODUCTION: The role of non-invasive positive pressure ventilation (NIPPV) in the treatment of acute lung injury (ALI) is controversial. We sought to assess the outcome of ALI that was initially treated with NIPPV and to identify specific risk factors for NIPPV failure. METHODS: In this observational cohort study at the two intensive care units of a tertiary center, we identified consecutive patients with ALI who were initially treated with NIPPV. Data on demographics, APACHE III scores, degree of hypoxemia, ALI risk factors and NIPPV respiratory parameters were recorded. Univariate and multivariate regression analyses were performed to identify risk factors for NIPPV failure. RESULTS: Of 79 consecutive patients who met the inclusion criteria, 23 were excluded because of a do not resuscitate order and two did not give research authorization. Of the remaining 54 patients, 38 (70.3%) failed NIPPV, among them all 19 patients with shock. In a stepwise logistic regression restricted to patients without shock, metabolic acidosis (odds ratio 1.27, 95% confidence interval (CI) 1.03 to 0.07 per unit of base deficit) and severe hypoxemia (odds ratio 1.03, 95%CI 1.01 to 1.05 per unit decrease in ratio of arterial partial pressure of O2 and inspired O2 concentration--PaO2/FiO2) predicted NIPPV failure. In patients who failed NIPPV, the observed mortality was higher than APACHE predicted mortality (68% versus 39%, p < 0.01). CONCLUSION: NIPPV should be tried very cautiously or not at all in patients with ALI who have shock, metabolic acidosis or profound hypoxemia. PMID- 16696864 TI - Prospective, randomized trial comparing fluids and dobutamine optimization of oxygen delivery in high-risk surgical patients [ISRCTN42445141]. AB - INTRODUCTION: Preventing perioperative tissue oxygen debt contributes to a better postoperative recovery. Whether the beneficial effects of fluids and inotropes during optimization of the oxygen delivery index (DO2I) in high-risk patients submitted to major surgeries are due to fluids, to inotropes, or to the combination of the two is not known. We aimed to investigate the effect of DO2I optimization with fluids or with fluids and dobutamine on the 60-day hospital mortality and incidence of complications. METHODS: A randomized and controlled trial was performed in 50 high-risk patients (elderly with coexistent pathologies) undergoing major elective surgery. Therapy consisted of pulmonary artery catheter-guided hemodynamic optimization during the operation and 24 hours postoperatively using either fluids alone (n = 25) or fluids and dobutamine (n = 25), aiming to achieve supranormal values (DO2I > 600 ml/minute/m2). RESULTS: The cardiovascular depression was an important component in the perioperative period in this group of patients. Cardiovascular complications in the postoperative period occurred significantly more frequently in the volume group (13/25, 52%) than in the dobutamine group (4/25, 16%) (relative risk, 3.25; 95% confidence interval, 1.22-8.60; P < 0.05). The 60-day mortality rates were 28% in the volume group and 8% in the dobutamine group (relative risk, 3.00; 95% confidence interval, 0.67-13.46; not significant). CONCLUSION: In patients with high risk of perioperative death, pulmonary artery catheter-guided hemodynamic optimization using dobutamine determines better outcomes, whereas fluids alone increase the incidence of postoperative complications. PMID- 16696865 TI - RIFLE criteria for acute kidney injury are associated with hospital mortality in critically ill patients: a cohort analysis. AB - INTRODUCTION: The lack of a standard definition for acute kidney injury has resulted in a large variation in the reported incidence and associated mortality. RIFLE, a newly developed international consensus classification for acute kidney injury, defines three grades of severity--risk (class R), injury (class I) and failure (class F)--but has not yet been evaluated in a clinical series. METHODS: We performed a retrospective cohort study, in seven intensive care units in a single tertiary care academic center, on 5,383 patients admitted during a one year period (1 July 2000-30 June 2001). RESULTS: Acute kidney injury occurred in 67% of intensive care unit admissions, with maximum RIFLE class R, class I and class F in 12%, 27% and 28%, respectively. Of the 1,510 patients (28%) that reached a level of risk, 840 (56%) progressed. Patients with maximum RIFLE class R, class I and class F had hospital mortality rates of 8.8%, 11.4% and 26.3%, respectively, compared with 5.5% for patients without acute kidney injury. Additionally, acute kidney injury (hazard ratio, 1.7; 95% confidence interval, 1.28-2.13; P < 0.001) and maximum RIFLE class I (hazard ratio, 1.4; 95% confidence interval, 1.02-1.88; P = 0.037) and class F (hazard ratio, 2.7; 95% confidence interval, 2.03-3.55; P < 0.001) were associated with hospital mortality after adjusting for multiple covariates. CONCLUSION: In this general intensive care unit population, acute kidney 'risk, injury, failure', as defined by the newly developed RIFLE classification, is associated with increased hospital mortality and resource use. Patients with RIFLE class R are indeed at high risk of progression to class I or class F. Patients with RIFLE class I or class F incur a significantly increased length of stay and an increased risk of inhospital mortality compared with those who do not progress past class R or those who never develop acute kidney injury, even after adjusting for baseline severity of illness, case mix, race, gender and age. PMID- 16696866 TI - Arteriolar vasoconstrictive response: comparing the effects of arginine vasopressin and norepinephrine. AB - INTRODUCTION: This study was designed to examine differences in the arteriolar vasoconstrictive response between arginine vasopressin (AVP) and norepinephrine (NE) on the microcirculatory level in the hamster window chamber model in unanesthetized, normotonic hamsters using intravital microscopy. It is known from patients with advanced vasodilatory shock that AVP exerts strong additional vasoconstriction when incremental dosage increases of NE have no further effect on mean arterial blood pressure (MAP). METHODS: In a prospective controlled experimental study, eleven awake, male golden Syrian hamsters were instrumented with a viewing window inserted into the dorsal skinfold. NE (2 microg/kg/minute) and AVP (0.0001 IU/kg/minute, equivalent to 4 IU/h in a 70 kg patient) were continuously infused to achieve a similar increase in MAP. According to their position within the arteriolar network, arterioles were grouped into five types: A0 (branch off small artery) to A4 (branch off A3 arteriole). RESULTS: Reduction of arteriolar diameter (NE, -31 +/- 12% versus AVP, -49 +/- 7%; p = 0.002), cross sectional area (NE, -49 +/- 17% versus AVP, -73 +/- 7%; p = 0.002), and arteriolar blood flow (NE, -62 +/- 13% versus AVP, -80 +/- 6%; p = 0.004) in A0 arterioles was significantly more pronounced in AVP animals. There was no difference in red blood cell velocities in A0 arterioles between groups. The reduction of diameter, cross sectional area, red blood cell velocity, and arteriolar blood flow in A1 to A4 arterioles was comparable in AVP and NE animals. CONCLUSION: Within the microvascular network, AVP exerted significantly stronger vasoconstriction on large A0 arterioles than NE under physiological conditions. This observation may partly explain why AVP is such a potent vasopressor hormone and can increase systemic vascular resistance even in advanced vasodilatory shock unresponsive to increases in standard catecholamine therapy. PMID- 16696867 TI - Early apoptosis of blood monocytes in the septic host: is it a mechanism of protection in the event of septic shock? AB - INTRODUCTION: Based on the central role of the triggering of monocytes for the initiation of the septic cascade, it was investigated whether apoptosis of blood monocytes in septic patients is connected to their final outcome. METHODS: Blood monocytes were isolated from 90 patients with septic syndrome due to ventilator associated pneumonia on days 1, 3, 5 and 7 from the initiation of symptoms. Apoptosis was defined after incubation with annexin-V-fluorescein isothiocyanate and propidium iodine and reading by a flow cytometer. The function of first-day monocytes was evaluated from the concentrations of tumour necrosis factor alpha (TNFalpha) and IL-6 in supernatants of cell cultures after triggering with endotoxins. TNFalpha, IL-6 and IL-8 were estimated in serum by an enzyme immunoassay. RESULTS: Mortality rates of patients with apoptosis < or =50% compared with patients with apoptosis >50% were 49.12% and 15.15%, respectively (P < 0.0001). Kaplan-Meier analysis showed a 28-day survival benefit in patients with septic shock and monocyte apoptosis >50% compared with those patients with apoptosis < or =50% (P = 0.0032). Production of IL-6 by monocytes on the first day by patients with apoptosis < or =50% was similar compared with monocytes isolated from healthy controls. Serum concentrations of TNFalpha were higher in patients with monocyte apoptosis < or =50% and septic shock compared with patients with apoptosis >50% on day 7; similar findings occurred for serum IL-6 on days 1 and 7 and for serum IL-8 on days 1 and 5. CONCLUSION: Early apoptosis of monocytes upon presentation of clinical signs of sepsis is connected to a favourable outcome. These findings are of particular importance for the patient with septic shock, where they might constitute a mechanism of pathogenesis. PMID- 16696868 TI - Periodontal disease and atherosclerosis from the viewpoint of the relationship between community periodontal index of treatment needs and brachial-ankle pulse wave velocity. AB - BACKGROUND: It has been suggested that periodontal disease may be an independent risk factor for the development of atherosclerosis. However, the relationship between periodontal disease and atherosclerosis has not been fully elucidated. This study aimed to assess the effects of periodontal disease on atherosclerosis. METHODS: The study design was a cross-sectional study. Subjects were 291 healthy male workers in Japan. We used the Community Periodontal Index of Treatment Needs (CPITN) score, average probing depth and gingival bleeding index (rate of bleeding gums) to assess the severity of periodontal disease. We also used the Brachial-Ankle Pulse Wave Velocity (baPWV) as the index for the development of atherosclerosis. RESULTS: The unadjusted odds ratio (OR) of atherosclerosis in relation to the CPITN score was 1.41 [95% CI: 1.16-1.73]. However, after adjustment for age, systolic blood pressure and smoking, the CPITN score had no relationship with atherosclerosis (adjusted OR: 0.91 [0.68-1.20]). CONCLUSION: Our results show no relationship between mild periodontal disease and atherosclerosis after appropriate adjustments. PMID- 16696869 TI - Induction of release and up-regulated gene expression of interleukin (IL)-8 in A549 cells by serine proteinases. AB - BACKGROUND: Hypersecretion of cytokines and serine proteinases has been observed in asthma. Since protease-activated receptors (PARs) are receptors of several serine proteinases and airway epithelial cells are a major source of cytokines, the influence of serine proteinases and PARs on interleukin (IL)-8 secretion and gene expression in cultured A549 cells was examined. RESULTS: A549 cells express all four PARs at both protein and mRNA levels as assessed by flow cytometry, immunofluorescence microscopy and reverse transcription polymerase chain reaction (PCR). Thrombin, tryptase, elastase and trypsin induce a up to 8, 4.3, 4.4 and 5.1 fold increase in IL-8 release from A549 cells, respectively following 16 h incubation period. The thrombin, elastase and trypsin induced secretion of IL-8 can be abolished by their specific inhibitors. Agonist peptides of PAR-1, PAR-2 and PAR-4 stimulate up to 15.6, 6.6 and 3.5 fold increase in IL-8 secretion, respectively. Real time PCR shows that IL-8 mRNA is up-regulated by the serine proteinases tested and by agonist peptides of PAR-1 and PAR-2. CONCLUSION: The proteinases, possibly through activation of PARs can stimulate IL-8 release from A549 cells, suggesting that they are likely to contribute to IL-8 related airway inflammatory disorders in man. PMID- 16696870 TI - Cognitive deficits and CTG repeat expansion size in classical myotonic dystrophy type 1 (DM1). AB - BACKGROUND: This study was designed to investigate cognitive abilities and their correlations with CTG repeat expansion size in classical Myotonic dystrophy type 1 (DM1), given that earlier studies have indicated cognitive deficits, possibly correlating with blood CTG repeats expansion size. METHODS: A measurement of CTG repeat expansion in lymphocytes and an extensive neuropsychological examination was made in 47 patients (25 women and 22 men). Individual results in the examination were compared with normative data. RESULTS: A substantial proportion of patients with DM1 (> 40%) scored worse in comparison to normative collectives on tests measuring executive, arithmetic, attention, speed and visuospatial abilities. We found significant correlations between longer CTG repeat expansion size and lower results on most tests associated with these abilities. Furthermore, the association between executive (frontal) deficits and DM1 were strengthened when considering both test results and correlations with CTG repeat expansion size in lymphocytes. CONCLUSION: This study showed deficits in several cognitive abilities when patients with DM1 are compared to normative collectives. Some of the neuropsychological tests associated with these abilities are correlated to CTG repeat expansion size in blood. These data highlight the importance of considering cognitive deficits when seeing patients with classical DM1 in clinical practice, but also the utility of using blood CTG repeat expansion size as a broad predictor of finding cognitive deficit in DM1. PMID- 16696871 TI - Prevalence of amyloid deposition in long standing rheumatoid arthritis in Iranian patients by abdominal subcutaneous fat biopsy and assessment of clinical and laboratory characteristics. AB - BACKGROUND: The study was aimed at determining the prevalence of secondary amyloidosis in a group of Iranian patients with Rheumatoid Arthritis (RA), and the assessment of its correlation with the clinical and laboratory findings and data. METHOD: A total number of 220 patients (167 female and 53 male) with a minimum five-year history of RA were selected. Congo red staining method was used for staining the specimens obtained by abdominal subcutaneous fat biopsy (ASFB) method. All of the specimens were examined for apple-green birefringence under polarized light microscope. Clinical and laboratory characteristics of the patients were assessed. Chi-square test and unpaired student's t-test were run for intergroup comparisons. RESULTS: Amyloid deposition test yielded positive results in 15 out of the 220 cases (6.8%) examined by the ASFB technique. Thirteen patients were found to have minimal amyloid deposits. Of all the clinically significant cases, 8 (53%) presented with proteinuria, and 7 cases (46.6%) had severe constipation. CONCLUSION: The prevalence of fat amyloid deposits in Iranian patients with RA is low. In up to half of the study group the deposits were subclinical. Follow up studies are required to determine whether this subclinical amyloidosis can develop into full-blown clinically significant amyloidosis. PMID- 16696872 TI - Obituary: Andrew William Morrison. PMID- 16696873 TI - Did Ludwig's angina kill Ludwig? AB - Wilhelm Frederick von Ludwig first described in 1836 a potentially fatal, rapidly spreading soft tissue infection of the neck and floor of the mouth. The condition was later named 'Ludwig's angina', a term which persists in medicine to this day. A gold medallist at 19 and professor at 25, Ludwig also served as president of the Wurttemberg Medical Association and chief physician to the royal family. His outstanding contribution to medicine was rewarded with the title Excellence upon retiring in 1855. Ludwig died at the age of 75, ironically, days after developing an inflammation of the neck. Could it be that Ludwig died of his own condition? This article combines a biography of Wilhelm Frederick von Ludwig with an overview of his eponymous condition and its management. PMID- 16696874 TI - Spatial orientation of the facial nerve in relation to parotid tumours. AB - OBJECTIVE: The aim of this study was to identify the distribution of parotid tumours within the gland in relation to the facial nerve branches. Documentation revealing such a relationship has not been reported previously. METHOD: A prospective study involving 111 patients was carried out over a period of 18 years in a specialist otolaryngology unit within a district general hospital. The relationship of the facial nerve to the tumour was graded into six types. The grading system was then employed to categorize each case. RESULTS: More than two thirds of the tumours were pleomorphic adenomas. The majority of these were located in the body and not in the tail of the parotid gland. In 50 per cent of these cases, the tumour was in close association with the two major divisions of the facial nerve. In the case of Warthin's tumours, 47 per cent were located in the tail, below the lower division of the facial nerve. PMID- 16696875 TI - Reliability of a tympanic thermometer in measuring temperatures in children after minor ear surgery. AB - AIM: To evaluate the reliability of infrared tympanic thermometry in children who have undergone myringotomy with grommet insertion. METHOD: Forty children who had undergone myringotomy with at least one grommet insertion had the tympanic temperature of each ear and the axillary temperature measured on admission and 30 minutes post-operatively. RESULT: No difference was found between the pre- and post-operative temperatures measured by either method (p > 0.05, paired t-test, hypothesized difference of 0). CONCLUSION: Infrared tympanic thermometry is reliable in monitoring body temperature in children who have had minor ear surgery. PMID- 16696876 TI - Hearing aid insertion: correlation between patients' confidence and ability. AB - INTRODUCTION: Once fitted with hearing aids, much reliance is placed on patients' own ability to manage their devices effectively. There has, however, been little research to assess how patients' own confidence compares with their actual ability to manage their hearing aids. This study compares patients' perceptions of their ability to insert their hearing aids with their observed ability to insert the devices satisfactorily. METHOD: Eighty-five patients provided a rating of their level of confidence in fitting their hearing aid, using a visual analogue score (VAS). This was then compared with their observed level of ability, assessed by an audiologist, also using a VAS (both scores 0-100 mm). RESULTS: We found a weak to moderate correlation between the subjective and objective scores (Pearson r = 0.4912). CONCLUSIONS: From these results, we would advise caution when accepting a patient's perceived level of ability as the only indicator of their true ability to insert their hearing aid. This also has important implications for follow-up services that rely only on telephone conversations with patients after a first fitting. PMID- 16696877 TI - Ear protection as a treatment for disruptive snoring: do ear plugs really work? AB - BACKGROUND: In the absence of surgical treatments with demonstrable long-term efficacy, conservative treatments for snoring remain appropriate. Only limited evidence evaluating the use of ear protection by partners has been published. METHODS: Eligible couples were invited to participate in a two-month trial, with each partner using custom-moulded ear plugs. Five questionnaires were completed by patients and their partners, at baseline and after two months: the snoring outcomes survey (SOS) or the spouse/bed partners survey (SBPS), the Epworth sleepiness scale (ESS), the general health questionnaire-12 (GHQ-12), the Nottingham health profile (NHP) and the Golombok-Rust inventory of marital status (GRIMS). RESULTS: After two months, there was a significant improvement in the SOS score for patients (p = 0.005), the SBPS score for partners (p = 0.001) and the ESS for partners (p = 0.004). CONCLUSIONS: For selected couples, ear plugs represent an effective short-term treatment for some of the social effects of disruptive snoring, as determined by the SOS, SBPS and ESS questionnaires. PMID- 16696878 TI - Optical quality of the nasendoscope with and without the endosheath. AB - With the increasing use of the endosheath in clinical practice, we set out to investigate the quality of the nasendoscope image produced with and without an endosheath. It has been suggested by some users that the endosheath degrades the image. We used a spectrophotometer to assess the optical transmission of the endosheath and found no selective chromatic absorption. However, on requesting nine experienced users to document whether they could differentiate between the sheathed and unsheathed endoscope, a significantly correct answering pattern was obtained in repeated blinded experiments. This suggests that individuals can distinguish between the sheathed and unsheathed nasendoscope view and that the image is indeed altered when an endosheath is employed. PMID- 16696880 TI - Laryngeal carcinoma: five-year survival and patterns of failure in 202 consecutive patients treated with primary or post-operative radiotherapy in Hong Kong. AB - OBJECTIVES: We aimed to conduct a retrospective analysis of patients treated with radiotherapy for laryngeal carcinoma at a single institution. METHODS: We analysed data from 202 consecutive patients treated with primary or post operative radiotherapy for laryngeal carcinoma over a 10-year period. RESULTS: Sixty-nine patients had a T1, 65 a T2, 39 a T3 and 29 a T4 lesion. Forty-one patients were node-positive. The clinical stage was I in 67 patients, II in 53, III in 36 and IV in 46. Primary radiotherapy was given to 152 patients. The median follow up was 60 months. The five-year overall local control rate was 86 per cent, the ultimate local control rate was 93 per cent, the five-year regional control rate was 96 per cent, the five-year relapse-free survival rate was 82 per cent and the five-year overall survival rate was 69 per cent. CONCLUSIONS: Patients with laryngeal carcinoma treated with primary or post-operative radiotherapy had a five-year overall survival rate of 69 per cent. PMID- 16696879 TI - Head and neck mucosal squamous cell carcinoma: results of palliative management. AB - OBJECTIVE: To document the outcome of patients deemed at initial diagnosis to be appropriate for palliative care. MATERIALS AND METHODS: Over a five-year period, all patients with head and neck mucosal squamous cell carcinoma (HNMSCC) were referred to one consultant surgeon, and data were collated retrospectively. RESULTS: Of 286 patients with HNMSCC, 60 (21 per cent) were deemed appropriate for palliative care at initial diagnosis. Their median survival was 154 days, with hypopharyngeal and oropharyngeal tumours forming a disproportional percentage of the initial diagnoses (p < 0.001). After initial assessment, 66 per cent of patients were discharged to their home, 17 per cent to hospice care and 17 per cent were transferred to other institutions. Including deaths during the initial admission, 29 per cent of patients did not return home after diagnosis. Palliative treatment included operative interventions in 35 per cent and radiotherapy in 43 per cent. Time spent in further hospital care averaged 47 days, and 47 per cent of patients eventually died in hospital. CONCLUSION: One in five patients presenting with HNMSCC were deemed appropriate for palliative care at initial presentation and survived less than six months after diagnosis. More than one-third required surgical intervention, and 29 per cent never returned home. PMID- 16696881 TI - Securing a paediatric airway with a nasal sucker for laser surgery. AB - The administration of anaesthetic gases to an infant undergoing an endolaryngeal laser procedure can be difficult. We describe the use of a Magill's nasal sucker and an anaesthetic connector to maintain anaesthesia during laser surgery. PMID- 16696882 TI - Angiolymphoid hyperplasia with eosinophilia of the auricle: progression of histopathological changes. AB - Angiolymphoid hyperplasia with eosinophilia is a rare condition that demonstrates dermal or subcutaneous proliferation of endothelial cells associated with an inflammatory cell infiltrate. A case is reported, with emphasis on the histopathological features on repeated biopsies. The report serves to stress the importance of considering this condition in the differential diagnosis of lesions in and around the ear. PMID- 16696883 TI - A bifid intra-tympanic facial nerve in association with a normal stapes. AB - A rare facial nerve anomaly was incidentally discovered whilst performing a tympanoplasty and ossicular reconstruction on a patient with an acquired unilateral conductive hearing loss. The nerve was seen to bifurcate and straddle a normal stapes superstructure as it ran posteriorly through the middle ear, a unique and as yet unreported combination. This case highlights the importance of vigilance regarding facial nerve anatomical variations encountered during middle ear surgery thus avoiding inadvertent damage. The purported embryological mechanism responsible for such anomalies of the intra-tympanic facial nerve is discussed. PMID- 16696884 TI - Deafness in Vogt-Koyanagi-Harada syndrome. AB - Sudden onset sensorineural hearing loss is a well recognized entity frequently encountered in otolaryngological practice. However, the combination of such deafness as part of a wider systemic disorder is fortunately rare. Almost 100 years after the syndrome was classified, we describe a case of Vogt-Koyanagi Harada syndrome occurring unusually in a Caucasian woman and characterized by sudden hearing loss. A brief review of this rare condition is presented. PMID- 16696885 TI - Bilateral vocal fold paralysis following treatment with vincristine. PMID- 16696888 TI - A case for full-coverage hard acrylic non-sleep-apnea dental orthotics. PMID- 16696889 TI - Perceptions and attitudes of Canadian dentists toward digital and electronic technologies. AB - OBJECTIVES: To determine dentists" perceptions of the usefulness of digital technologies in improving dental practice and resolving practice issues; to determine dentists" willingness to use digital and electronic technologies; to determine perceived obstacles to the use of digital and electronic technologies in dental offices; and to determine dentists" attitudes toward Internet privacy issues. METHODS: An anonymous, self-administered survey of Canadian dentists was conducted by mail. A potential mailing list of 14,052 active Canadian dentists was compiled from the 2003 records of provincial regulatory bodies. For each province, 7.8% of the dentists were randomly selected with the help of computer software. The surveys were mailed to this stratified random sample of 1,096 dentists. RESULTS: The response rate was 28% (312/1,096). Of the 312 respondents, 4 (1%) were in full-time academic positions, 16 (5%) were not practising, and 9 (3%) provided incomplete data. Therefore, 283 survey responses were available for analysis. More than 60% of the dentists indicated that computer technology was quite capable or very capable of improving their current practice by increasing patient satisfaction, decreasing office expenses, increasing practice efficiency, increasing practice production, improving record quality and improving case diagnosis and treatment planning. More than 50% of respondents reported that digital photography and digital radiography were quite useful or very useful. About 70% of the dentists agreed or strongly agreed with using digital and electronic technologies to consult with dental specialists. Cost of equipment and lack of comfort with technology were regarded as significant or insurmountable obstacles by substantial proportions of respondents. CONCLUSIONS: Respondents generally viewed digital and electronic technologies as useful to the profession. Increased office efficiency and production were perceived as positive effects of digital and electronic technologies. These technologies are more often used for consulting with colleagues rather than for consulting with patients. The major obstacles to the general use of these technologies were related to cost, lack of comfort with technology and differences in legislation between provinces and countries. Privacy issues were not perceived as a significant barrier. PMID- 16696890 TI - Comparative analysis of microleakage and seal for 2 obturation materials: Resilon/Epiphany and gutta-percha. AB - OBJECTIVE: Microleakage continues to be a main reason for failure of root canal therapy, where the challenge has been to achieve an adequate seal between the internal tooth structure and the main obturation material, gutta-percha. The Resilon/Epiphany (R/E) system uses a new obturation material that bonds chemically with the internal tooth structure, thereby decreasing the possibility of microleakage. The purpose of this study was to compare dye leakage in root canals filled with R/E and those in which gutta percha was used. METHODS AND MATERIALS: Pulpectomies were performed on 105 extracted human singlecanal mandibular incisors. The teeth were then randomly divided into 2 groups: 1 was obturated with gutta-percha and the other with R/E. Obturation was performed using the lateral condensation technique. The teeth were then incubated for 10 days, 1 month or 3 months in a heated water bath solution with dye to simulate conditions in the human oral cavity. Teeth were sectioned and examined under dissecting and scanning electron microscopes to assess dye penetration, seal and bonding. RESULTS: Resilon as the main obturation material consistently resulted in less microleakage than gutta-percha at all 3 time intervals. CONCLUSION: The R/E system provides a new material for root canal treatment. R/E creates a chemical bond with the internal tooth structure over the entire root area that is maintained over time, thus representing a better option than gutta-percha. Further studies on R/E will help validate its use and determine its long-term success rates in vivo. PMID- 16696891 TI - Persuasive evidence that formocresol use in pediatric dentistry is safe. AB - Concern has been expressed about the safety of formocresol use in pediatric dentistry. Formaldehyde, a primary component in formocresol, is a hazardous substance and is considered a probable human carcinogen by Health Canada. However, humans inhale and ingest formaldehyde daily and also produce this compound as part of normal cellular metabolism. The human body is physiologically equipped to handle this exposure through multiple pathways for oxidation of formaldehyde to formate and incorporation into biological macromolecules via tetrahydrofolate-dependent one-carbon biosynthetic pathways. Recent re-evaluation of earlier research that examined potential health risks associated with formaldehyde exposure has shown that the research was based on flawed assumptions, which resulted in erroneous conclusions. This review examines more recent research about formaldehyde metabolism, pharmacokinetics and carcinogenicity, the results of which indicate that formaldehyde is probably not a potent human carcinogen under conditions of low exposure. Extrapolation of these research results to pediatric dentistry suggests an inconsequential risk of carcinogenesis associated with formaldehyde use in pediatric pulp therapy. Areas for further investigation are suggested. PMID- 16696892 TI - Transmigration of impacted mandibular canines--report of 4 cases. AB - Impacted canines are not uncommon in clinical practice, but intraosseous movement of impacted canines crossing the midline (transmigration) is a rare phenomenon. We report 4 cases of mandibular canine transmigration to emphasize the need to supplement periapical radiographs with a panoramic radiographic examination in patients with over-retained deciduous canines or missing permanent canines. PMID- 16696893 TI - Longevity of a maxillary 2-unit cantilever fixed partial denture: clinical report. AB - In this clinical report, we discuss the length of service and subsequent replacement of a maxillary anterior 2-unit cantilever fixed partial denture (FPD). The FPD provided 53 years of service to the patient and was finally replaced with a 2-unit porcelain-fused-to-metal (PFM) FPD. The original prosthesis replaced a missing maxillary lateral incisor using a partial coverage metal retainer, whereas the new FPD was designed with a complete coverage PFM retainer. PMID- 16696894 TI - Kinetics of receptor-ligand interactions in immune responses. AB - Receptor-ligand interactions in blood flow are crucial to initiate the biological processes as inflammatory cascade, platelet thrombosis, as well as tumor metastasis. To mediate cell adhesions, the interacting receptors and ligands must be anchored onto two apposing surfaces of two cells or a cell and a substratum, i.e., the two-dimensional (2D) binding, which is different from the binding of a soluble ligand in fluid phase to a receptor, i.e., three-dimensional (3D) binding. While numerous works have been focused on 3D kinetics of receptor-ligand interactions in immune systems, 2D kinetics and its regulations have less been understood, since no theoretical framework and experimental assays have been established until 1993. Not only does the molecular structure dominate 2D binding kinetics, but the shear force in blood flow also regulates cell adhesions mediated by interacting receptors and ligands. Here we provided the overview of current progresses in 2D bindings and regulations. Relevant issues of theoretical frameworks, experimental measurements, kinetic rates and binding affinities, and force regulations, were discussed. PMID- 16696895 TI - The T-box transcription factor T-bet in immunity and autoimmunity. AB - The T-box transcription factor T-bet (Tbx21) has emerged as a key regulator of type 1-like immunity, playing critical roles in the establishment and/or maintenance of effector cell fates in T and B lymphocytes, as well as dendritic cells and natural killer cells. Several autoimmune diseases, especially those classically considered related to T helper 1 (Th1) immunity, appear to require T bet, at least as judged in mouse models. This review summarizes a current understanding of T-bet's role in immunity, as well as its importance in autoimmunity, with implications for therapeutic intervention. PMID- 16696896 TI - The immune response induced by hepatitis B virus principal antigens. AB - Hepatitis B virus (HBV) infection occurs primarily in hepatocytes in the liver with release of infectious virions and non-infectious empty surface antigen particles into the bloodstream. HBV replication is non-cytopathic. Transient infections run a course of several months, and chronic infections are often life long. Chronic infections can lead to liver failure with cirrhosis and hepatocellular carcinoma. It is generally accepted that neutralizing anti-HBs antibodies plays a key role in recovery from HBV infection by containing the spread of infection in the infected host and facilitating the removal and destruction of viral particles. However, the immune response initiated by the T cell response to viral antigens is also important for viral clearance and disease pathogenesis in HBV infection. The three structural forms of the viral proteins, the HBsAg, the particulate HBcAg, and the nonparticulate HBeAg, may preferentially elicit different Th cell subsets. The different IgG subclass profiles of anti-HBs, anti-HBc, and anti-HBe in different HBV infection status were revealed. Moreover, the different IgG subclass profiles in chronic carriers did not change with different ALT and AST levels and may reflect the difference between stimulating antigens, immune response, and the stages of viral disease and provide the basis for the use of vaccines and prophylactic treatments for individuals at high risk of human HBV infection. This review elucidates the detailed understanding of the immune responses induced during transient and persistent infection, and the development of immunotherapy and immunodiagnosis in patients with HBV infection, and possible means of reducing the liver damage. PMID- 16696897 TI - Target organ protection from a novel angiotensin II receptor (AT1) vaccine ATR12181 in spontaneously hypertensive rats. AB - Hypertension produces pathophysiological changes that are often responsible for the mortality associated with the disease. It is evident that overactive renin angiotensin systems play a central role in the development of hypertension and target organ damage associated with hypertension. We have previously found that a novel angiotensin II receptor (AT1) vaccine-ATR12181 attenuated the development of high blood pressure (BP) in spontaneously hypertensive rat (SHR) model of human essential hypertension. Our objective was to determine whether this attenuation of high BP is associated with prevention of target organ damage induced by hypertensive state. SHRs were immunized against a peptide (coded ATR12181) from the extracelluar portion of the AT1A receptor by repeated subcutaneous injections of peptide-tetanus-toxoid complex in combination with Freund's adjuvant. A 64 weeks long-term observation was performed. Repeated vaccinations resulted in the induction of anti-ATR12181 antibodies. At the end of observation, vaccinated SHRs manifested lower BP, decreased cardiac hypertrophy and attenuation of kidney injuries. mRNA levels of c-fos and c-jun in heart and kidneys were decreased in vaccinated SHRs. Since a self antigen was used, safety of vaccine was concerned. However, the signs of autoimmune diseases were not observed in the sections of heart and kidney. These data demonstrated that repeated immunization against a domain of the extracellular portion of the AT1 receptor was able to cause a target organ protection against hypertension. Active immunization against the AT1 receptor may be considered as a promising new strategy in the treatment of hypertension. PMID- 16696898 TI - Toxin(s), other than cholera toxin, produced by environmental non O1 non O139 Vibrio cholerae. AB - A total of 39 Vibrio cholerae non O1 non O139 strains were isolated from surface waters of different parts of Dhaka City, Bangladesh. All these strains showed lack of ctx or zot gene, as demonstrated by the PCR analysis. Eighteen representative strains were tested for enterotoxin production using a rabbit ileal loop model, of which live cells of 8 strains and culture filtrates of 6 strains produced fluid accumulation in ileal loops. However, none of them produced heat stable toxin (ST), as detected by suckling mouse assay. On the other hand, 15% of isolates produced cytotoxin as detected by the Chinese Hamster Ovary (CHO) cell assay. Fifty times concentrated culture filtrates of the representative strains did not give any precipitin band against the anti-cholera toxin, suggesting the strains produced an enterotoxin, which is antigenically different from known cholera toxin (CT). Eighty percent of the total isolates were found to be positive for heat labile haemolysin detected by tube method, whereas, 39% were found positive by the Christie-Atkins-Munch-Petersen (CAMP) method. However, 87% of the isolates were positive for haemagglutinin/protease and all of the strains were positive for mannose-sensitive-haemagglutinin assay. PMID- 16696899 TI - The ex vivo microenviroments in MLTC of poorly immunogenic tumor cells facilitate polarization of CD4+CD25+ regulatory T cells. AB - CD4+CD25+ regulatory T (TR) cells play an important role in maintaining a balanced peripheral immune system. Recent studies have shown that TR cells may also play a key role in suppressing anti-tumor immune response. In order to investigate the tumor immune microenvironment and its influence on TR polarization, poorly immunogenic tumor cell line D5 (C57BL/6, H-2b), immunogenic tumor cell lines FBL3 (C57BL/6, H-2b) and H22 BALB/c, H-2d) were used to establish the syngeneic/allogeneic, poorly immunogenic/immunogenic mixed lymphocytes-tumor cell culture (MLTC). Our results revealed that the proportion of CD4+CD25+ T cells in MLTC of syngeneic primed splenocytes stimulated with D5 tumor cells was higher than that with H22 cells (0.43% vs 0.044%, and the similar results appeared in allogeneic splenocytes stimulated with D5 tumor cells (0.39% vs 0.04%). The splenocytes stimulated with supernatant from syngeneic MLTC of D5 tumor cells demonstrated higher proportion of CD4+CD25+ cells than that from allogeneic MLTC of D5 tumor cells, and the splenocytes stimulated with supernatant from syngeneic or allogeneic MLTC of H22 tumor cells generated lower proportion of CD4+CD25+ T cells than that of D5 tumor cells. The TGF-beta1 and Th2-oriented cytokines (IL-4 and IL-10) were dominated in supernatants of syngeneic MLTC of poorly immunogenic tumor cells. Our results provided useful information for studying the mechanisms underlying tumor immune surveillance as well as for the tumor immunotherapy. PMID- 16696900 TI - Increased uterine NK-derived IFN-gamma and TNF-alpha in C57BL/6J mice during early gestation. AB - Natural killer (NK) cells are bone marrow-derived lymphocytes. They produce cytokines that regulate the development of acquired immunity. In view of their accumulation at the maternal-fetal interface, uterine natural killer (uNK) cells are also thought to play essential roles during pregnancy. Our results compared the differences of cytokine secretion profile by NK cells in uterine endometrium, liver, spleen and peripheral blood, and focused on the cytokines secretion by uNK cells. It was demonstrated that the expression of IFN-gamma and TNF-alpha in uterine endometrium of pregnant mice are lower than those in liver, but they increase significantly during pregnancy. Our study showed that the number of uNK cells was increased significantly during pregnancy. They produced more IFN-gamma and TNF-alpha than other organ-derived NK cells, and they also secreted minor amount of IL-4 and IL-5. The results indicated that the IFN-gamma and TNF-alpha produced by uNK cells ensured a successful pregnancy progress. PMID- 16696901 TI - Expression and purification of soluble human programmed death-1 in Escherichia coli. AB - Programmed death-1 (PD-1), a member of CD28 family, is able to negatively regulate the TCR complex-initiated signaling by interacting with its cognate ligands (PD-L1 and/or PD-L2). PD-1/PD-L1 pathway plays an important role in down regulating the effective phase of adaptive immune responses and the blockade of this pathway has been proved to enhance antiviral and antitumoral immunity, suggesting that it might be a potential target for the development of therapies to improve T cell responses in patients with virus infections or malignancies. In present study, the extracellular domain of human PD-1 with a carboxyl terminal His-tag (designated as sPD-1) was expressed as inclusion bodies in Escherichia coli. The product was on-column refolded, purified by immobilized metal affinity chromatography, and characterized by Western blotting. Furthermore, the soluble PD-1 with high purity possessed specific binding activity with its cognate ligand PD-L1, and the dissociation constant was 0.43 nmol/L as determined by Scatchard plot analysis. These results suggest that refolded sPD-1 from prokaryotic cells may be of therapeutic interest in enhancing antivirus and antitumoral immune responses. PMID- 16696902 TI - Different characters of spleen OX-62 positive dendritic cells between Fischer and Lewis rats. AB - The phenotype, DNA-binding activities of NF-kappaB, cytokine production, endocytosis and stimulatory capacity of spleen OX-62-positive dendritc cells (SDCs) from Fischer rats were compared with those from Lewis rats. Results showed that the expressions of CD11b, MHC-II, CD8, CD45RA, CD54 and CD86 on SDCs were significantly higher in Fischer than those in Lewis rats. The levels of IL-2, IL 4, IL-10 and IFN-gamma in SDCs from Fischer rats were distinctly higher than those from Lewis. Both stimulatory capacity and DNA-binding activities of NF kappaB in SDCs were all lower in Fischer than those in Lewis rats. These differences may partly contribute to rat strain-specificity in susceptibility to chronic inflammatory stimuli. PMID- 16696903 TI - Blood serum levels of IL-2, IL-6, IL-8, TNF-alpha and IL-1beta in patients on maintenance hemodialysis. AB - Cytokines are essential mediators of immune response and inflammatory reactions. Patients with chronic renal failure (CRF) commonly present with abnormalities of immune function related with impaired kidney function and the accumulation of uremic toxins in addition to bioincompatibility of dialyzer membranes. During a hemodialysis (HD) session, cytokines are released mainly by monocytes activated by endotoxin-type compounds in dialyzer fluid, complement factors and direct contact with dialyzer membrane. The study included 15 CRF patients, aged 36.4 +/- 2.9 years, on regular HD maintenance therapy for mean 68 +/- 10 months and 15 healthy controls. It was designed to assess serum levels of a panel of inflammatory cytokines: IL-1beta, IL-2, IL-6, IL-8 and TNF-alpha in CRF patients on regular maintenance HD before, 20, 60 and 240 minutes of a single HD session in parallel with C-reactive protein (CRP) as an additional parameter. CRP concentration was increased in HD patients when compared with healthy controls. The concentrations of IL-1, IL-6, IL-8 and TNF-alpha were increased, whereas the serum level of IL-2 was not altered during a single HD session. PMID- 16696904 TI - MS in Asian countries. AB - This paper reviews all available English language literature on MS from Asian countries published between 1970 and 2005. Although limited data are available, the review reveals that western Asia--including the Middle East--has the highest prevalence of MS across the continent, and that MS in Asia largely resembles conventional MS in western countries. Opticospinal MS (a distinct clinical entity from conventional MS) is more common in eastern Asian regions. Larger epidemiological and genetic studies, with more complete ascertainment in various Asian populations, are needed so that we can understand the diversity of Asian MS. PMID- 16696905 TI - [Research direction for syndromes of traditional Chinese medicine: differentiating diseases from syndromes and differentiating syndromes from diseases]. AB - By historical review on integrative research of syndrome differentiation and disease differentiation and considering the scientific research method of traditional Chinese medicine (TCM), the syndrome is the key subject of clinical study on TCM. During the recent 10 years or more, many new diseases and diagnostic approaches have been found and developed in Western medicine, while in TCM, the advances in research on syndrome can not keep up with the requirements of clinical practice. This problem was caused by three matters as follows: (1) Syndrome differentiation and its classification have been conservative and lack of new discovery; (2) The thinking pattern of clinical practice has been focused on disease differentiation, and only complemented by syndrome differentiation; (3) The treatment has been concentrated on the disease instead of on the syndrome. Considering these problems, the author has put forward the thinking pattern of clinical research, that is "to differentiate diseases from syndromes and to differentiate syndromes from diseases". In this way, many new syndromes and diseases will be discovered with the improvement of disease and syndrome differentiations. These advances will provide rich resources for basic theoretical and pharmacal studies of TCM, and improve the common understanding of TCM in the world. PMID- 16696906 TI - Improving the quality of randomized controlled trials in Chinese herbal medicine, part III: quality control of Chinese herbal medicine used in randomized controlled trials. AB - OBJECTIVE: To discuss quality control of Chinese herbal medicine (CHM) in randomized controlled trials (RCTs), and to provide suggestions for improving this aspect in future clinical study in this therapeutic area. METHODS: A search of the Cochrane Library was conducted to identify RCTs of CHM. Quality control information reported in those RCTs was then assessed independently. RESULTS: The search yielded a total of 167 RCTs of CHM for a variety of conditions. A total of 11 CHM preparations were used in those RCTs. Only one trial discussed quality control of the CHM interventions used. Issues affecting the safety and efficacy of CHM products used in RCTs were discussed including standardization of raw herbal materials, processing methods, screening for product contamination, and effects of combination products. CONCLUSION: The overall quality of reporting of RCTs of CHM was poor, reflecting the need for improvements in reporting future clinical trials in this area. RECOMMENDATIONS: To improve quality control of CHM used in RCTs in future, we recommend developing and implementing guidelines such as Good Agricultural Practice (GAP) for Chinese crude drugs, and current Good Manufacturing Practice (GMP) specific to CHM products. Chemical analyses of individual herbs of CHM and combination products are also recommended to provide reference standards for quality control. PMID- 16696907 TI - Improving the quality of randomized controlled trials in Chinese herbal medicine, part IV: applying a revised CONSORT checklist to measure reporting quality. AB - OBJECTIVE: To discuss the quality of reporting in randomized controlled trials (RCTs) of Chinese herbal medicine (CHM), and to provide suggestions for improving the reporting of future clinical studies in this therapeutic area. METHODS: A search of the Cochrane Library was conducted to identify RCTs of CHM. A revised CONSORT checklist designed for CHM clinical studies was implemented. The revised CONSORT checklist contained 63 items, including the following new items added specifically for CHM: (1) "syndrome of disease" based on Chinese medicine theories; (2) rationale of CHM formula; (3) formula composition; (4) preparation form of CHM; (5) quality control of CHM. RESULTS: The overall reporting quality of the RCTs as assessed with the revised CONSORT checklist varied between 19% and 44%, with a median score of 32% (standard deviation 8%). CONCLUSION: The overall quality of reporting of RCTs of CHM evaluated with a revised CONSORT checklist was poor, reflecting the need for improvements in reporting future clinical trials in this area. RECOMMENDATIONS: To improve the quality of reporting of RCTs of CHM, we recommend adopting a revised CONSORT checklist that includes items specific to CHM. We also recommend that editors of CHM journals require authors to use a structured approach to presenting their trials as a condition of publication. PMID- 16696908 TI - [A neglected master on integrative medicine: Wei-Ju ZHU]. AB - Through analyzing the academic ideas in Shanghan Zhinan written by Wei-Ju ZHU, a modern famous physician, it is pointed out that ZHU's achievement on integrative medicine has been overlooked. Taking the theory of syndrome differentiation of six channels from Shanghan Lun as an example, ZHU presumed that the essence of therapeutic theory of traditional Chinese medicine was to conform to or strengthen human body's self-healing and self-regulating abilities. Since Yangqi is the major force in the body to help heal disease when it occurred, ZHU often took measures to strengthen Yang first in treating disease and also was good at using hot-natured herbs, such as Radix aconiti lateralis preparata. ZHU's thoughts on integrative medicine were concerned with etiology, pathology and therapeutics. In addition, ZHU generalized the syndrome differentiation into the theory of five phases and eight principles, and made important contributions to the history of integrative medicine. His thinking will provide valuable revelation for clinical practice and modern research on traditional Chinese medicine. PMID- 16696909 TI - [Effects of yisui shengxue granules on expressions of alpha-hemoglobin stabilizing protein and erythroid transcription factor GATA-1 mRNAs in bone marrow of patients with beta-thalassemia]. AB - OBJECTIVE: To evaluate the effects of Yisui Shengxue Granules on expressions of alpha-hemoglobin stabilizing protein (AHSP) and erythroid transcription factor GATA-1 mRNAs in bone marrow of patients with beta-thalassemia, and to explore its possible molecular mechanism. METHODS: Twelve patients with beta-thalassemia intermedia were treated with Yisui Shengxue Granules for three months. The blood indexes including hemoglobin (Hb), RBC, fetal hemoglobin (HbF) and reticulated corpuscles (Ret) were examined before and after treatment. Total RNA was extracted from bone marrow karyocyte in 8 patients selected from these 12 patients before and after treatment, and the expression levels of the AHSP and GATA-1 mRNAs were measured by real-time PCR. RESULTS: Yisui Shengxue Granules could not only obviously improve the clinical symptoms of patients with beta thalassemia intermedia, but also obviously increased the contents of Hb, RBC, HbF and Ret (P<0.05, or P<0.01). The expression levels of AHSP and GATA-1 mRNAs also significantly increased after treatment as compared with those before treatment (P<0.05, or P<0.01). CONCLUSION: The results revealed that one of the possible molecular mechanism of the effects caused by Yisui Shengxue Granules is that it can up-regulate the expression levels of AHSP and erythroid transcription factor GATA-1 mRNAs, enhance the protein synthesis of AHSP which can bind the relative excess free alpha-globin, prevent the formation of alpha -globin-cytotoxic precipitates in red blood cells and decrease the hemolysis. PMID- 16696910 TI - [Effects of xuezhikang capsules on vascular endothelial function and redox status in patients with coronary heart disease]. AB - OBJECTIVE: To explore the effects of Xuezhikang Capsules (ZXKC) and probucol on blood lipids, vascular endothelial functions and redox status in patients with coronary heart disease. METHODS: One hundred and twelve patients with coronary heart disease were randomly divided into XZKC-treated group and probucol-treated group, 56 in each. Before and after 8-week treatment, the blood levels of total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL C), nitric oxide (NO), endothelin-1 (ET-1), reduced glutathione (GSH) and oxidized glutathione (GSSG) were all measured in both groups. The GSH/GSSG redox potential (Eh) was calculated according to the Nernst equation. RESULTS: In the XZKC-treated group, the blood levels of TC, LDL-C and TG were significantly decreased after 8-week treatment as compared with those before treatment. The blood levels of TC and LDL-C were also significantly decreased in the probucol treated group as compared with those before treatment. In the XZKC-treated group, the blood levels of ET-1 and GSSG and the GSH/GSSG Eh after treatment were all significantly lower than those before treatment, whereas the blood levels of GSH and NO, the NO/ET-1 ratio, and the GSH/GSSG ratio after treatment were all significantly higher than those before treatment. CONCLUSION: The XZKC or probucol treatment can yield a significant decrease in blood lipids in patients with coronary heart disease. Furthermore, XZKC exerts effective protection on vascular endothelial function, and can make GSH/GSSG redox status shift towards deoxidation. PMID- 16696911 TI - [Effects of xuefu zhuyu decoction on functions of vascular endothelium in patients with unstable angina pectoris]. AB - OBJECTIVE: To evaluate the effects of Xuefu Zhuyu Decoction (XFZYD) on functions of vascular endothelium in patients with unstable angina pectoris (UAP). METHODS: Fifty patients with UAP were randomly divided into two groups: XFZYD-treated group with 32 cases and control group with 18 cases. The patients in the control group were given regular therapy, while the patients in the XFZYD-treated group were given XFZYD additionally on the basis of regular therapy. All patients in both groups were treated for 8 weeks. Then the contents of serum endothelin (ET), nitric oxide (NO), soluble vascular cell adhesive molecule-1 (sVCAM-1) and soluble intercellular adhesion molecule-1 (sICAM-1) were examined by radioimmunoassay, enzymic method and enzyme-linked immunosorbent assay (ELISA) respectively. RESULTS: The total response rate in the XFZYD-treated group was 93.75%, which was significantly higher than 66.67% in the control group (P<0.05). The contents of serum ET, sVCAM-1 and sICAM-1 in the XFZYD-treated group were much lower than those in the control group (P<0.01), while the content of serum NO was obviously higher than that in the control group (P<0.01). CONCLUSION: XFZYD can improve the functions of vascular endothelium by lowering the levels of endothelium-derived contracting substances, enhancing the levels of endothelium derived relaxing substances, and reducing the cell adhesions, and hence to raise the therapeutic effects on UAP. PMID- 16696912 TI - [Volt-ampere characteristics of Yuan points of three yin channels of hand in women before, during and after menstruation]. AB - OBJECTIVE: To observe the changes of volt-ampere characteristics of Yuan-Primary acupoints in women before, during and after menstruation, and to find out the relationships between Qi-blood in human body and volt-ampere characteristics of acupoints, so as to develop a new method for quantitative analysis on changes of Qi-blood. METHODS: A high-sensitive detection system of volt-ampere characteristics of acupoint was applied to detecting the volt-ampere characteristics of Taiyuan (LU 9), Daling (PC 7) and Shenmen (HT 7) in healthy women before, during and after menstruation. RESULTS: The resistances on LU 9, PC 7 and HT 7 with some certain scan currents were quite different among the women before, during and after menstruation. For such changes, PC 7 was of the most obvious change, LU 9 was of the less and HT 7 was of the least. The total ratio of the scanning spots with significantly increasing changes of resistances on these three acupoints along with the menstrual process was 70.6%. CONCLUSION: The volt-ampere characteristics of acupoints vary with the menstrual process, which is of the acupoint specificity. The resistances on the acupoints increase with the menstrual process, and such results may be due to the blood loss during menstruation. The volt-ampere characteristics can be used as a quantitative index to study on the change of Qi and blood. PMID- 16696913 TI - [Establishment of a multidrug-resistant cell line BEL-7402/5-FU of human hepatocellular carcinoma and its biological characteristics]. AB - OBJECTIVE: To establish a multidrug-resistant cell line BEL-7402/5-FU of hepatocellular carcinoma (HCC). METHODS: BEL-7402/5-FU was induced by pulse therapy combined with continuous stepwise exposure to 5-fluorouracil in vitro. MTT assay was used to determine its multidrug resistance (MDR). Biological characteristics of the BEL-7402/5-FU cell line were observed including morphological changes, cell growth curve, population doubling time, plate cloning efficiency, adherence rate, cell cycle distribution, chromosome and tumorigenicity. Accumulation amount of adriamycin (ADM) in cytoplasm was measured by flow cytometry. The protein expression of thymidylate synthase (TS) was evaluated by immuno-cytochemical method. RESULTS: The acquired MDR cell line of BEL-7402/5-FU was established successfully. The BEL-7402/5-FU cells showed cross resistance to ADM, vincristine (VCR), methotrexate (MTX) and oxaliplatin (OHP), whereas still sensitive to hydroxycamptothecin (HCPT). The BEL-7402/5-FU cells tended to grow in clusters in vitro. It was found that the population doubling time of BEL-7402/5-FU cells was longer than that of its parental cells. The plate cloning efficiency and the adherence rate of BEL-7402/5-FU cells at the 2nd and 3rd hour were both lower than those of the parental cells. The distributing proportion of BEL-7402/5-FU cells in G(0)/G(1) phase was less than that of the parental cells, whereas the distributing proportion of BEL-7402/5-FU cells in S phase was higher than that of the parental cells. The accumulation amount of ADM in cytoplasm of BEL-7402/5-FU cells was significantly lower while the expression level of TS protein of which was highly up-regulated as compared with those of the parental cells. CONCLUSION: Establishment of the human HCC cell line BEL 7402/5-FU might be beneficial to the studies of 5-Fluorouracil acquired MDR mechanisms and the selection of reversal modifiers. PMID- 16696914 TI - [Effects of fuzheng yiliu granules on apoptotic rate and mitochondrial membrane potential of hepatocellular carcinoma cell line H22 from mice]. AB - OBJECTIVE: To evaluate the effects of Fuzheng Yiliu Granules (FZYLG) on apoptotic rate and mitochondrial membrane potential (Delta psi m) of hepatocellular carcinoma cell line H22 from mice. METHODS: Forty-eight mice inoculated with H22 cells were randomly divided into four groups: untreated group, cyclophosphamide treated group, high-dose FZYLG-treated group and low-dose FZYLG-treated group. After 14 days of corresponding treatment, H22 cells in each group were stained with propidium iodide, and the apoptotic rates were detected by flow cytometry (FCM). The rhodamine 123 was used as a fluorescence probe to label the H22 cells, and the fluorescence intensities were observed with laser scanning confocal microscope. The fluorescence intensity of H22 cells indicated the Delta psi m of H22 cells. RESULTS: FZYLG could significantly increase the apoptotic rate while reduce the Delta psi m of H22 cells from mice as compared with those in the untreated group. CONCLUSION: The antitumor effects of FZYLR on H22 cells from mice are related to decreasing the Delta psi m and then inducing the apoptosis of the H22 cells. PMID- 16696915 TI - [Effects of jinlongshe granules on apoptosis of MKN-45 human gastric cancer cells orthotopically transplanted in nude mice]. AB - OBJECTIVE: To assess the effects of Jinlongshe Granules (JLSG) on tumor growth of gastric carcinoma. METHODS: Fifty nude mice orthotopically transplanted with MKN 45 human gastric cancer cells were divided into five groups: untreated group, 5 fluorouracil (5-FU)-treated group and high-, medium-, and low-dose JLSG-treated groups. Corresponding antitumor drugs were administered in each group except the untreated group. The antitumor effects in vivo were evaluated. Cell cycle distribution and apoptosis of MKN-45 human gastric cancer cells were determined by using flow cytometry (FCM) and Annexin V-fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI) staining assay. The ultrastructure of MKN-45 gastric cancer cells was observed by transmission electron microscope. RESULTS: In the mice treated with high-, medium-, and low-dose JLSG, the growth inhibition rates of gastric cancer were 68.13%, 55.94% and 50.31% respectively, and this antitumor effect was dose-dependent. In the mice treated with intraperitoneal injection of 5-FU, the growth inhibition rate of gastric cancer was 53.43% and not much different from those treated with JLSG. The apoptotic rates in the high-, medium , and low-dose JLSG-treated groups were 22.81%, 28.27% and 38.54% respectively, in a dose-dependent manner, with the cell cycle arrested at G(0)/G(1) phase. An Annexin V-FITC/PI staining assay revealed that the percentages of early apoptotic cells in the three dose JLSG-treated groups were all significantly higher than that in the 5-FU-treated group, whereas the late apoptotic and necrotic cells were much more in the 5-FU-treated group than those in the three dose JLSG treated groups. CONCLUSION: Jinlongshe Granules exert an inhibiting effect on MKN 45 human gastric cancer cell. PMID- 16696916 TI - [Establishment of a rat model of alcoholic liver fibrosis induced by complex factors]. AB - OBJECTIVE: To establish a model of alcoholic liver fibrosis (ALF) in rats induced by complex factors. METHODS: Forty-seven healthy male rats were divided into three groups: normal control group (n=12), minor CCl4 group (n=12) and complex factors group (n=27). The rats in the complex factors group were fed a complex diet including alcohol, corn oil and pyrazole, and administered with intraperitoneal injection of minor CCl4 to induce ALF. During induction process, the histopathological changes of liver tissue and the values of liver-to-body weight ratio were both observed regularly. The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma glutamyltransferase (gamma-GT) in these three groups were all examined at the 12th week of the induction process. RESULTS: At the 12th week of the induction process, the model of ALF induced by complex factors was successfully established in rats, and the histopathological presentations showed alcoholic fatty liver, hepatitis and liver fibrosis in a sequence along with the induction process. The value of liver-to-body weight ratio and the serum levels of ALT, AST and gamma-GT of rats in the complex factors group were all significantly different from those in the other two groups. CONCLUSION: It is a steady and effective way to induce ALF in rats with complex diet and minor CCI4 injection. PMID- 16696917 TI - [Inhibiting effects of root of Mallotus apelta on duck hepatitis B virus]. AB - OBJECTIVE: To evaluate the inhibiting effects of the root of Mallotus apelta (Lour.) Muell.-Arg. on duck hepatitis B virus (D-HBV) in vivo. METHODS: Forty nestling ducks with congenitally infection of D-HBV detected by PCR were randomly divided into five groups: untreated group, lamivudine-treated group, and high-, medium- and low-dose root of Mallotus apelta-treated groups. The ducks in the lamivudine-treated group were fed lamivudine with a dose of 50 mg/kg once. Ducks in the three-dose Mallotus apelta-treated groups were treated with different doses of decoction of this herbal medicine for 21 days respectively. The serum content of D-HBV DNA was determined by quantitative real-time PCR technique before and 7 days after the treatment, and on the 7th, 14th and 21st day of the treatment. Liver biopsy was also executed before and after the treatment to observe the histopathological changes. RESULTS: Lamivudine showed a rapid inhibiting effect on D-HBV DNA, but this effect didn't last long, and the serum level of D-HBV DNA increased again after treatment. The serum level of D-HBV DNA dropped markedly in the high- and medium-dose Mallotus apelta-treated groups on the 14th and 21st day. Low-dose Mallotus apelta revealed no obvious inhibiting effect on D-HBV. After treatment, the inhibiting effect in the root of Mallotus apelta-treated group continued as compared with that in the untreated group. The histopathological changes of liver tissues showed that the inflammation in the high-dose root of Mallotus apelta-treated group was weakened as compared with that in the lamivudine-treated group. CONCLUSION: The root of Mallotus apelta has therapeutic effect on D-HBV. It can restrain the duplication of D-HBV in vivo. Although this effect is weaker than that of lamivudine, it continues longer. Thus this herbal medicine is an effective, safe and economical drug for hepatitis B. PMID- 16696918 TI - [Effects of shenshuning recipe on expression of hepatocyte growth factor in rats with glomerulosclerosis]. AB - OBJECTIVE: To investigate the effects of Shenshuning Recipe (SSNR) on gene expression of hepatocyte growth factor (HGF) in renal tissues in rats with glomerulosclerosis. METHODS: Glomerulosclerosis was induced in 42 rats by unilateral nephrectomy and intravenous injection of doxorubicin. Then these 42 rats were randomly divided into three groups: untreated group, SSNR-treated group and benazepril-treated group. Another eight rats were included into sham operation group. The rats in the SSNR-treated group and the benazepril-treated group were fed SSNR or benazepril respectively for 8 weeks. The levels of 24 h urine protein (Upr), serum creatinine (Cr) and blood urea nitrogen (BUN) of rats in each group were examined. The renal morphological changes were observed under microscope, and the diameter of glomerular capillary, mesangial matrix and glomerulosclerosis index were analyzed by image analysis software. Reverse transcription-polymerase chain reaction (RT-PCR) method was used to detect the gene expression of HGF in the renal tissues. RESULTS: The levels of 24 h Upr, serum Cr and BUN in the untreated group were remarkably increased than those in the sham-operation group (P<0.01). The pathological morphological changes in the untreated group showed that the glomerulosclerosis was diffused around the renal tissue and the capillaries were shrunk. The expression level of mesangial matrix was up-regulated and the glomerulosclerosis index was 3.32+/-0.35. The expression level of HGF mRNA in the untreated group was obviously lower than that in the sham-operation group (P<0.05). The levels of 24 h Upr, serum Cr and BUN in the SSNR-treated group and the benazepril-treated group were remarkably decreased as compared with those in the untreated group, while the expression levels of HGF mRNA were both obviously higher than that in the untreated group (P<0.01). The pathological morphological changes in the SSNR-treated group and the benazepril treated group were both alleviated. There was no significant difference in therapeutic effect between the SSNR-treated group and the benazepril-treated group. CONCLUSION: Shenshuning Recipe can up-regulate the expression of HGF mRNA, decrease the mesangial matrix, and improve the renal function, so that it may retard the development of glomerulosclerosis. PMID- 16696919 TI - [An in vitro natural degeneration model of chondrocytes derived from endplate of intervertebral discs of rats]. AB - OBJECTIVE: To set up a natural degeneration model of chondrocytes derived from endplate of intervertebral discs of rats in order to offer an appropriate carrier for the study on mechanism of intervertebral disc degeneration. METHODS: The method of enzyme digestion combined with natural subculture was used to set up the in vitro natural degeneration model of chondrocytes derived from the endplate of intervertebral disc of rats. The morphological appearances and microstructures of the chondrocytes of different generations were observed. The expression of collagen II in chondrocytes was detected by immunocytochemical method. RESULTS: The chondrocytes derived from the endplate of intervertebral disc expressed collagen II. After 13 days of culture, the chondrocytes of generation III showed that the ability of cell division descended, the nucleoli became unclear, the cells deformed obviously, fusiform shape with weak optical activity appeared, and the intercellular space was enlarged. There were vacuoles and lipid droplets in cytoplasm. The synthesis of collagen II, as well as the cell proliferation rate, descended notably. All results showed the natural degeneration process of the chondrocytes. CONCLUSION: The in vitro natural degeneration model of chondrocytes derived from endplate of intervertebral discs of rats was successfully established. This can offer the cytological basis for study on the mechanism of intervertebral disc degeneration. PMID- 16696920 TI - [Primary study on proteomics about Ganoderma lucidium spores promoting survival and axon regeneration of injured spinal motor neurons in rats]. AB - OBJECTIVE: To detect some proteins associated with the effect of ganoderma lucidium spores (GASP) on promoting the survival and axon regeneration of injured spinal motor neurons in rats. METHODS: The rats were divided into normal control group, untreated group and GASP-treated group, and the rats in the last two groups received ventral root avulsion. GASP preparation was fed to the rats in the GASP-treated group for 14 days. The gray matter tissues of the lumbar spinal were sampled from rats in each group after 14 days following ventral root avulsion, and the extracted proteins from these tissues were detected by using 2 dimensional electrophoresis. Matrix assisted laser desorption/ionization time-of flight mass spectroscopy (MALDI-TOF MS) was utilized to identify the differentially expressed proteins among these three groups. RESULTS: There were six kinds of proteins differentially expressed among the three groups, which were collapsin response mediator protein 2 (CRMP-2), F-actin capping protein beta subunit (FCP-beta), isocitrate dehydrogenase [NAD] subunit beta (IDH-beta), ATPase, glutamate oxaloacetate transaminase-1 (GOT1) and M2 pyruvate kinase (M2 PK). The expression levels of CRMP-2, IDH-beta, ATPase and GOT1 were higher in the GASP-treated group than those in the untreated group, while the expression levels of FCP-beta and M2-PK were lower than those in the untreated group. CONCLUSION: GASP maybe promotes the survival and axon regeneration of injured spinal motor neurons in rats by virtue of up- or down-regulating the expression levels of the proteins mentioned above. PMID- 16696921 TI - [Effects of soybean isoflavones on expression levels of osteoprotegerin and osteoprotegerin ligand mRNAs in bone tissues of ovariectomized rats]. AB - OBJECTIVE: To investigate the effects of soybean isoflavones (SI) on expression levels of osteoprotegerin (OPG) and osteoprotegerin ligand (OPGL) mRNAs in bone tissues of ovariectomized rats, and to discuss the mechanism of soybean isoflavaones in preventing and treating postmenopausal osteoporosis (PMO). METHODS: Thirty adult SD rats were randomly divided into 3 groups: sham-operation group, untreated group and SI-treated group. The rats in the last two groups were bilaterally ovariectomized. The bone density of L(3) to L(6) vertebrae was detected after 12-week intervention. Total RNA was extracted from femur head and the expression levels of OPG and OPGL mRNAs were examined by real-time quantitative polymerase chain reaction. RESULTS: SI could increase the bone density of lumbar vertebrae in ovariectomized rats, up-regulate the expression level of OPG mRNA and down-regulate the ratio of OPGL mRNA/OPG mRNA, but exert no significant effect on the expression of OPGL mRNA. CONCLUSION: The therapeutic effects of SI on PMO may be related to regulating the expression levels of OPG and OPGL mRNAs, and the ratio of OPGL mRNA/OPG mRNA. PMID- 16696922 TI - [Effects of kangfengshi granules on expressions of osteoprotegerin, RANKL and M CSF in bone tissues of rats with collagen-induced arthritis]. AB - OBJECTIVE: To observe the effects of Kangfengshi Granules (KFSG) on expressions of the mRNAs of osteoprotegerin (OPG), receptor activator of nuclear factor kappaB ligand (RANKL) and macrophage colony stimulating factor (M-CSF) in bone tissues of rats with collagen-induced arthritis. METHODS: Forty SD rats were randomly divided into four groups: normal control group, untreated group, cyclosporine A (CsA)-treated group and KFSG-treated group. Except the rats in the normal control group, all the other rats received subcutaneous injection of collagen II to establish collagen-induced arthritis (CIA) models. Then the rats in each group were fed normal saline or corresponding drugs for four weeks. Total RNA was extracted from carpal and digital bones. The expressions of OPG, RANKL and M-CSF mRNAs were examined by real-time PCR. RESULTS: The total incidence of arthritis induced by collagen II in the rats was approximately 90%. The expression levels of RANKL and M-CSF mRNAs and the RANKL mRNA/OPG mRNA ratio in the untreated group, KFSG-treated group and CsA-treated group were all significantly higher than those in the normal control group, while the expression levels of OPG mRNA in those three groups were significantly lower than that in the normal control group. The expression level of OPG mRNA in the KFSG-treated group was obviously higher while the expression level of M-CSF mRNA and the RANKL mRNA/OPG mRNA ratio in the same group were both lower as compared with those in the untreated group. CONCLUSION: The molecular mechanism of effects of KFSG on bone erosion and destruction induced by rheumatoid arthritis is closely correlated with up-regulating the expression of OPG mRNA, down-regulating the expression of M-CSF mRNA and RANKL mRNA/OPG mRNA ratio. PMID- 16696923 TI - [Research on real-time data collection and demonstration system for acupuncture manipulation]. AB - OBJECTIVE: To apply the real-time data collection and demonstration system for acupuncture manipulation to quantitative analysis of acupuncture manipulation for acupuncture teaching and research. METHODS: The real-time observation and analysis of parameter changes of acupuncture manipulation were implemented by this system. RESULTS: During the course of lifting and thrusting manipulation, the needle body moved in a uniform variable motion, and the action force was still kept while the arrival of Qi was achieved. During the course of twirling manipulation, the variations of twirling number and radian caused the changes of resistance moment, and this reaction was related to the body response. During the course of rotating and shaking manipulation, the radius for rotating and shaking was related to such factors as the force, the action point and the direction. CONCLUSION: The real-time data collection and demonstration system for acupuncture manipulation can provide a new experimental method for quantitative data analysis, standardized research and teaching demonstration of acupuncture. PMID- 16696924 TI - [Clinical observation on treatment of gastric cancer with zhijing granules: a report of 148 cases]. PMID- 16696925 TI - [Clinical effects of tonglin huoxue decoction on patients with ureteral calculi]. PMID- 16696926 TI - [Experiences in treatment based on syndrome differentiation for black fur caused by dampness stagnancy due to spleen deficiency]. PMID- 16696927 TI - [Progress in treatment of diabetes mellitus and complications by Chinese herbal drugs]. PMID- 16696928 TI - [Personal idea on English translation of traditional Chinese medicine: theory and practice]. PMID- 16696929 TI - Relationship between mechanisms and activities at the time of pedestrian injury and activity limitation among school adolescents in Kathmandu, Nepal. AB - This study assessed the relationship between pedestrian activity at the time of injury, the type of vehicle involved and resulting activity limitation among school adolescents in the Kathmandu and Lalitpur districts of Nepal. A cross sectional study of 1557 students in grades 6-8 across 14 schools was conducted using a self-administered questionnaire from August to September 2003. Twenty three percent of adolescents reported pedestrian injuries, 38% were from urban and 21% from semi-urban areas. Adolescents were commonly injured by motorcycles and motor vehicles while crossing the road; however, while walking and playing, they were commonly injured by bicycles and motorcycles. Bicycles and motor vehicles were less likely to be involved in injury while crossing the roads and playing, respectively (p < 0.001). Activity was more likely to be limited for a longer period of time (> 7 days) with injuries endured while crossing the road (p < 0.001). In urban areas, boys and girls were more likely to be injured while crossing the road and walking, respectively (p < 0.05), and both were commonly injured by motorcycles. In semi-urban areas, boys and girls were commonly injured while walking and were more likely to be injured by motorcycles and bicycles, respectively (p < 0.05). In both areas, more boys than girls were injured while playing. These findings have important implications for pedestrian safety interventions in poor countries. PMID- 16696930 TI - Factors associated with automobile accidents and survival. AB - This paper develops an econometric model for vehicles' inherent mortality rate and estimates the probability of accidents and survival in the United States. Logistic regression model is used to estimate probability of survival, and censored regression model is used to estimate probability of accidents. The estimation results indicated that the probability of accident and survival are influenced by the physical characteristics of the vehicles involved in the accident, and by the characteristics of the driver and the occupants. Using restrain system and riding in heavy vehicle increased the survival rate. Middle aged drivers are less susceptible to involve in an accident, and surprisingly, female drivers are more likely to have an accident than male drivers. Riding in powerful vehicles (high horsepower) and driving late night increase the probability of accident. Overall, the driving behavior and characteristics of vehicle does matter and affects the probabilities of having a fatal accident for different types of vehicles. PMID- 16696931 TI - Improving pilots' risk assessment skills in low-flying operations: the role of feedback and experience. AB - Risk assessment is one of the most important skills that pilots are expected to acquire to ensure the safe and successful management of flight. The traditional approach to the development of these skills requires pilots to directly engage with potentially hazardous events. Using low-flying as a context, the present study sought to test whether engagement with hazards in a simulated environment, together with feedback concerning performance, would improve pilots' risk assessment during a subsequent simulated test flight. The results indicated that engagement with the hazards, rather than the provision of feedback per se, was associated with behavior that reduced the risk to the aircraft, while maintaining operational performance. It was concluded that exposure to hazards within a simulated environment could provide the basis for the development of risk assessment skills amongst less experienced pilots. PMID- 16696932 TI - Experimental model of chronic osteomyelitis caused by Leishmania (L) amazonensis. AB - Experimental animal models have been used for the study of the physiopathogenesis of leishmaniasis, on some occasions with success, while in other situations such as bone alterations that accompany tegumentary leishmaniasis, especially in diffuse cutaneous form (DCL), the mechanisms are still unknown. In the present study, we determined these alterations in an animal model susceptible to Leishmania (L) amazonensis. Amastigotes of L. (L) amazonensis isolated from patients with diffuse cutaneous leishmaniasis (DCL) were inoculated into the hind paws of eight BALB/c mice, macroscopic and histopathological aspects were analyzed. After 90 and 120 days of evolution, histopathological analysis demonstrated a mononuclear cell infiltrate rich in plasma cells and intense parasitism of intra- and extra-medullary macrophages, with areas of bone necrosis and discrete involvement of cartilaginous tissue. The results show that the inflammatory process developed during L. (L) amazonensis infection might cause bone tissue destruction and secondarily affect the joints. PMID- 16696933 TI - Competency-based training for patient handling. AB - A technique-training approach has traditionally been used to address the problem of back pain associated with patient handling. This project aimed to investigate whether different levels of safety culture, based on competency-based training, resulted in different behaviour (physical and cognitive) for patient handling tasks. Sixteen healthcare organisations in the UK participated from the acute and primary healthcare sectors. Archival data for each organisation were benchmarked against the Royal College of Nursing competencies for manual handling. Behavioural data were collected on two patient handling tasks: (1) sitting-to standing and (2) repositioning-in-sitting using observations (postural analysis) and interviews (verbal protocol analysis). The data were analysed for each organisation and then grouped by task and method into larger data sets. These data sets were triangulated using the key decision-making points (from the interview data) as the framework. The results showed that in organisations with a more positive safety culture the nursing staff demonstrated more complex decision making about the patient handling tasks and had lower levels of associated postural risk. PMID- 16696934 TI - Homogeneous boron targeting of heterogeneous tumors for boron neutron capture therapy (BNCT): chemical analyses in the hamster cheek pouch oral cancer model. AB - OBJECTIVE: BNCT is a tumour cell targeted radiation therapy. Uniform targeting of heterogeneous tumours with therapeutically effective boron carriers would contribute to a therapeutic effect on all tumour cell populations and avoid radioresistant fractions. This remains an unresolved challenge. The aim of the present study was to evaluate the degree of variation in boron content delivered by boronophenylalanine (BPA), GB-10 (Na(2)(10)B(10)H(10)) and the combined administration of (BPA+GB-10) in different portions of tumour, precancerous tissue around tumour and normal pouch tissue in the hamster cheek pouch oral cancer model. MATERIALS AND METHODS: Samples of different areas of tumour, precancerous tissue and normal pouch tissue were taken from tumour-bearing hamsters, 3h post-administration of i.p. BPA (15.5mg B/kg b.w.), or i.v. GB-10 (50mg B/kg b.w.), or 3h and 1.5h post-administration respectively of i.v. GB-10 (34.5mg B/b.w.) and sequential i.p. injections of BPA (total dose 31mg B/kg b.w.) given jointly. Boron content was evaluated by inductively coupled plasma optical emission spectroscopy (ICP-OES). The degree of homogeneity in boron targeting was assessed in terms of the coefficient of variation (V: [S.D./mean]x100) of boron values. Statistical analysis of the results was performed by one-way ANOVA and the least significant difference test. RESULTS: GB-10 and GB-10 plus BPA achieved respectively a statistically significant 1.8- and 3.3-fold increase in targeting homogeneity over BPA. CONCLUSIONS: The combined boron compound administration protocol contributes to homogeneous targeting of heterogeneous tumours and would be expected to increase therapeutic efficacy of BNCT. PMID- 16696935 TI - High-resolution electrophysiology on a chip: Transient dynamics of alamethicin channel formation. AB - Microstructured planar substrates have been shown to be suitable for patch clamp recording from both whole cells and isolated patches of membrane, as well as for measurements from planar lipid bilayers. Here, we further explore this technology with respect to high-resolution, low noise single-channel recording. Using solvent-free lipid bilayers from giant unilamellar vesicles obtained by electro swelling, we recorded channels formed by the peptaibol alamethicin, a well studied model system for voltage-dependent channels, focusing on the transient dynamics of single-channel formation upon application of a voltage step. With our setup, we were able to distinctly resolve dwell times well below 100 mus and to perform a thorough statistical analysis of alamethicin gating. Our results show good agreement with models that do not rely on the existence of non-conducting preaggregate states. Microstructured apertures in glass substrates appear promising with respect to future experiments on cellular ion channels reconstituted in suspended lipid membranes. PMID- 16696936 TI - Regulation of cholesterol 25-hydroxylase expression by vitamin D3 metabolites in human prostate stromal cells. AB - Vitamin D3 plays an important role in the control of cell proliferation and differentiation. Cholesterol 25-hydroxylase (CH25H) is an enzyme converting cholesterol into 25-hydroxycholesterol. Vitamin D3 as well as 25 hydroxycholesterol has been shown to inhibit cell growth and induce cell apoptosis. Here we show that 10 nM 1alpha,25(OH)2D3 and 500 nM 25OHD3 upregulate CH25H mRNA expression in human primary prostate stromal cells (P29SN). Protein synthesis inhibitor cycloheximide does not block 1alpha,25(OH)2D3 mediated upregulation of CH25H mRNA. Transcription inhibitor actinomycin D blocks basal level as well as 1alpha,25(OH)2D3 induced CH25H mRNA expression. 1alpha,25(OH)2D3 has no effect on CH25H mRNA stability. 25-Hydroxycholesterol significantly decreased the P29SN cell number. A CH25H enzyme inhibitor, desmosterol, increases basal cell number but has no significant effect on vitamin D3 treated cells. Our data suggest that ch25h could be a vitamin D3 target gene and may partly mediate anti-proliferative action of vitamin D3 in human primary prostate stromal cells. PMID- 16696937 TI - Mutation analysis of Taiwanese Wilson disease patients. AB - Wilson disease (WD) is an autosomal recessive disorder of copper metabolism, which is caused by mutation in copper-transporting ATPase (ATP7B). In the present study, we report a molecular diagnosis method to screen the WD chromosome in patients or in heterozygotic carriers in Taiwan. Exons 8, 11, 12, 13, 16, 17, and 18 of ATP7B are selected for the screening of mutations. The most common mutation, Arg778Leu or Arg778Gln, was first screened by PCR-RFLP then we combined single-stranded conformation polymorphism (SSCP) analysis followed by direct DNA sequencing on the DNA fragments with mobility shift on SSCP analysis. The diagnostic rate was compared with standard ATP7B whole gene sequencing analysis. Ten different mutations were identified among 29 WD patients; among them four were novel (Ala1168Pro, Thr1178Ala, Ala1193Pro, and Pro1273Gln). The false positive rates were tested against 100 normal individuals and listed as follows: exon 8: 5%; exon 11: 4%; exon 12: 6%; exon 13: 5%; exon 16: 5%; exon 17: 3%; exon 18: 4%. The Arg778Leu mutation exhibited the highest allelic frequency (43.1%). The detection rate of WD chromosomes is 65.52%, which is as sensitive as whole gene sequencing scanning. According to our results, WD chromosomes in Taiwan are predominantely located at exons 8, 11, 12, 13, 16, 17, and 18. The standard sequencing analysis on the entire gene is time consuming. We recommend screening these 7 exons first on those individuals who have a higher risk in having WD, before whole gene and promoter sequencing analysis in Taiwan. PMID- 16696938 TI - IFN-alpha induces apoptosis of adipose tissue cells. AB - Interferon alpha (IFN-alpha) is produced in response to viral infections and used clinically in the therapy of a variety of cancers and viral infections. IFN-alpha treatment is often associated with severe weight reduction. To elucidate the mechanism of IFN-associated weight loss, we studied its effect on adipocytes in vitro and in vivo. Diet-induced obese (DIO) C57BL/6 mice were treated continuously for 8 days with human IFN-alpha A/D (100 U/g body weight) or with vehicle alone. The body weight and adipose cell size of IFN-alpha A/D-treated DIO mice were significantly lower (P<0.05 and P<0.001, respectively) as compared with those of control DIO mice. PI3K and Bcl-2 were down-regulated whereas Bax expression was elevated in adipose tissue following IFN treatment as compared to adipose tissue of control DIO mice. Treatment of differentiated 3T3-F442A adipocytes with IFN-alpha A/D (250 U/ml, 36 h) significantly increased the number of apoptotic cells from 15.8% in control cells to 56+/-6%. In conclusion, weight loss following IFN-alpha therapy is due at least in part to increased apoptosis of adipocytes. PMID- 16696939 TI - Acarbose, an alpha-glucosidase inhibitor, improves endothelial dysfunction in Goto-Kakizaki rats exhibiting repetitive blood glucose fluctuation. AB - Several epidemiological studies have revealed that subjects with postprandial hyperglycemia are at increased risk of cardiovascular disease. However, the impact of postprandial hyperglycemia and its treatment on endothelial function has not been clarified yet. In this study, Goto-Kakizaki (GK) rats, a non-obese type 2 diabetes model, fed twice daily were used as a model of repetitive postprandial glucose spikes. We investigated the endothelial function in these rats treated or untreated with acarbose, an alpha-glucosidase inhibitor. Administration of acarbose for 12 weeks markedly improved postprandial hyperglycemia, postprandial insulin level, total cholesterol, triglyceride, and free fatty acid level in GK rats. Furthermore, acarbose efficiently reduced the number of monocytes adherent to aortic endothelial layer, improved acetylcholine dependent vasodilatation, and reduced intimal thickening of the aorta. While it is generally regarded that repetitive postprandial hyperglycemia is associated with the onset of cardiovascular diseases, our data demonstrated that acarbose treatment efficiently ameliorated endothelial dysfunction and reduced intimal thickening, thus adding support to the protective effect of acarbose against the onset of cardiovascular disease. PMID- 16696940 TI - High glucose increases B1-kinin receptor expression and signaling in endothelial cells. AB - The loss of endothelial function is the initiating factor in the development of diabetic vascular disease. Kinins control endothelial function by the activation of two receptors: the B2 which is constitutively expressed, and the B1 which is highly induced in pathological conditions. In the present study, we observed that the levels of B1-receptor mRNA and protein are induced in endothelial cells incubated in high glucose. An increase in B1-receptor was also observed in the endothelial layer of aortas, from 4-week diabetic rats. When cells were grown in high glucose, the B1 agonist des-Arg9-BK increased nitrite levels, whereas in normal glucose nitrite levels were unchanged. Nitrite increase was blocked by L NAME and 1400W indicating the participation of the inducible Nitric Oxide Synthase (iNOS). iNOS protein levels were also increased in high glucose. These results demonstrate the participation of the B1 receptor in the signaling pathways mediated by kinins in high glucose. PMID- 16696941 TI - Functional cross-talk between p73beta and NF-kappaB mediated by p300. AB - p73beta is associated with induction of apoptosis or cellular growth arrest, while NF-kappaB is closely related with promotion of resistance to programmed cell death. These biologically opposing activities between p73beta and NF-kappaB propose a regulatory mechanism of critical turning on/off in cellular apoptotic or survival responses. In this study, we demonstrate that NF-kappaB-mediated transactivation is specifically downregulated by p73beta; conversely, p73beta transactivation is negatively regulated by functional expression of p65, NF kappaB RelA subunit. The p73beta transactivation domain (TA) and p65 NH2-terminus are crucial for their negative regulation of p65- and p73beta-mediated transactivation, respectively. Furthermore, p65- or p73beta-interaction with p300 is reciprocally inhibited by their competitive binding to p300 in a restrict amount-dependent manner. Likewise, both p73beta-activated apoptosis and p65 dependent increase of cell viability are reciprocally repressed by p65 and p73beta, respectively. These results have important implications for p300 mediated regulatory mechanism between p73beta- and p65-transactivation, by which both p73beta and NF-kappaB could mutually affect on their biological activities. Therefore, we propose that p300 is a transactivational regulator of competitively balanced cross-talk between p73beta and p65. PMID- 16696942 TI - UVA-mediated down-regulation of MMP-2 and MT1-MMP coincides with impaired angiogenic phenotype of human dermal endothelial cells. AB - UVA irradiation, dose-dependently (5-20 J/cm2), was shown to impair the morphogenic differentiation of human microvascular endothelial cells (HMECs) on Matrigel. Parallely, UVA down-regulated the expression of MMP-2 and MT1-MMP, both at the protein and the mRNA levels. On the contrary, the production of MMP-1 and TIMP-1 by HMECs increased following UVA treatment. The inhibitory effect of UVA on MMP expression and pseudotubes formation was mediated by UVA-generated singlet oxygen (1O2). The contribution of MT1-MMP, but not TIMP-1, to the regulation of HMECs' angiogenic phenotype following UVA irradiation was suggested using elastin derived peptides and TIMP-1 blocking antibody, respectively. PMID- 16696943 TI - O2-mediated oxidation of hemopexin-heme(II)-NO. AB - Hemopexin (HPX), serving as scavenger and transporter of toxic plasma heme, has been postulated to play a key role in the homeostasis of NO. Here, kinetics of HPX-heme(II) nitrosylation and O2-mediated oxidation of HPX-heme(II)-NO are reported. NO reacts reversibly with HPX-heme(II) yielding HPX-heme(II)-NO, according to the minimum reaction scheme: HPX-heme(II)+NO kon<-->koff HPX heme(II)-NO values of kon, koff, and K (=kon/koff) are (6.3+/-0.3)x10(3)M-1s-1, (9.1+/-0.4)x10(-4)s-1, and (6.9+/-0.6)x10(6)M-1, respectively, at pH 7.0 and 10.0 degrees C. O2 reacts with HPX-heme(II)-NO yielding HPX-heme(III) and NO3-, by means of the ferric heme-bound peroxynitrite intermediate (HPX-heme(III)-N(O)OO), according to the minimum reaction scheme: HPX-heme(II)-NO+O2 hon<--> HPX heme(III)-N(O)OO l-->HPX-heme(III)+NO3- the backward reaction rate is negligible. Values of hon and l are (2.4+/-0.3)x10(1)M-1s-1 and (1.4+/-0.2)x10(-3)s-1, respectively, at pH 7.0 and 10.0 degrees C. The decay of HPX-heme(III)-N(O)OO (i.e., l) is rate limiting. The HPX-heme(III)-N(O)OO intermediate has been characterized by optical absorption spectroscopy in the Soret region (lambdamax=409 nm and epsilon409=1.51x10(5)M-1cm-1). These results, representing the first kinetic evidence for HPX-heme(II) nitrosylation and O2-mediated oxidation of HPX-heme(II)-NO, might be predictive of transient (pseudo-enzymatic) function(s) of heme carriers. PMID- 16696944 TI - A novel cis-acting element in Her2 promoter regulated by Stat3 in mammary cancer cells. AB - Stat3 plays important roles in the development of breast malignancies and oncogenesis. In the present study, a palindromic cis-acting element displaying repression activity in breast cancer cells expressing low level of Her2 was found in Her2 promoter. Deletion analysis showed that the novel element was located within Pal2 region spanning nucleotides -529 to -505. The sequence analysis of Pal2 region revealed a DNA sequence (TTAAGATAA) homologous to the binding site of Stat3, starting from position -529 to -521bp. By reporter assay, Pal2 was found to be regulated by constitutive activated Stat3C. A stimulatory effect both on Her2 mRNA and protein expressions was observed in MCF-7 cells stably expressing Stat3C, suggesting that Stat3 regulated Her2 expression. Using ChIP assays the binding of Stat3 to Her2 promoter was confirmed. The data obtained in this study indicate constitutive activated Stat3 regulates Her2 expression. Further investigation of differential effects of Stat3 exerting on breast cancer cells expressing Her2 at different levels will provide more insights into the roles of Stat3 in Her2 expression as well as the regulation of diverse biological activities. PMID- 16696945 TI - Activation and roles of ALK4/ALK7-mediated maternal TGFbeta signals in zebrafish embryo. AB - Activin, Nodal, and Vg1, members of the transforming growth factor beta (TGFbeta) superfamily, transduce signal through type I receptors ALK4 or ALK7 and play important roles in mesoderm induction and patterning during vertebrate embryogenesis. However, the timing and magnitude of the ALK4/ALK7-mediated maternal TGFbeta signals are not clear. SB-431542 is identified as an inhibitor of the ALK4/ALK5/ALK7-mediated TGFbeta signals and its specificity in vertebrate embryos has not been reported. We demonstrate that SB-431542 is able to specifically and reproducibly block the Smad2/3-mediated TGFbeta signals in zebrafish embryo. Embryos exposed to SB-431542 exhibit various defects phenocopying Nodal-deficient mutants. SB-431542 treatments starting at different cell cycles before the midblastula transition lead to different degrees of developmental defects in mesoderm induction and patterning, suggesting that maternal TGFbeta signals are activated right after fertilization and required for mesoderm formation and patterning. PMID- 16696946 TI - Age-dependent oxidative stress-induced DNA damage in Down's lymphocytes. AB - The aim of the present study was to investigate the oxidative status of lymphocytes from children (n=7) and adults (n=18) with Down's syndrome (DS). The basal oxidative condition, the vulnerability to in vitro hydrogen peroxide exposure, and the repair capacity were measured by means of the damage-specific alkaline comet assay. Significantly and age-independently elevated numbers of single strand breaks and oxidized bases (pyrimidines and purines) were found in the nuclear DNA of the lymphocytes in the DS group in the basal condition. These results may support the role of an increased level of endogenous oxidative stress in DS and are similar to those previously demonstrated in Alzheimer's disease. In the in vitro oxidative stress-induced state, a markedly higher extent of DNA damage was observed in DS children as compared with age- and gender-matched healthy controls, suggesting that young trisomic lymphocytes are more sensitive to oxidative stress than normal ones. However, the repair ability itself was not found to be deteriorated in either DS children or DS adults. PMID- 16696948 TI - Suppression of CaCYP1, a novel cytochrome P450 gene, compromises the basal pathogen defense response of pepper plants. AB - A putative cytochrome P450 gene from chili pepper, Capsicum annuum L. Bukang cytochrome P450 (CaCYP1), was identified using cDNA microarray analysis of gene expression following induction of the leaf hypersensitive response by inoculation of pepper plants with the non-host pathogen Xanthomonas axonopodis pv. glycines 8ra. The full-length cDNA of CaCYP1 encoded a protein of 514 amino acid residues, which contained a putative hydrophobic membrane anchoring domain in the N terminal region, and a heme-binding motif in the C-terminal region. Analysis of the deduced amino acid sequence of CaCYP1 revealed that it has high homology to Arabidopsis CYP89A5, the function of which is unknown. Expression of CaCYP1 was preferentially increased in pepper plants in response to non-host pathogen inoculation and also during the host resistance response. CaCYP1 expression also increased following treatment with salicylic acid and abscisic acid, while treatment with ethylene had a mild effect. Using a virus-induced gene silencing based reverse genetics approach, we demonstrated that suppression of CaCYP1 results in enhanced susceptibility to bacterial pathogens. Interestingly, gene silencing of CaCYP1 in pepper plants resulted in the reduced expression of the defense-related genes CaLTP1, CaSIG4, and Cadhn. Our results indicated that CaCYP1, a novel cytochrome P450 in pepper plants, may play a role in plant defense response pathways that involve salicylic acid and abscisic acid signaling pathways. PMID- 16696947 TI - Phosphodiesterase 8 (PDE8) regulates chemotaxis of activated lymphocytes. AB - The immune system depends on chemokines to recruit lymphocytes to tissues in inflammatory diseases. This study identifies PDE8 as a new target for inhibition of chemotaxis of activated lymphocytes. Chemotactic responses of unstimulated and concanavalin A-stimulated mouse splenocytes and their modulation by agents that stimulate the cAMP signaling pathway were compared. Dibutyryl cAMP inhibited migration of both cell types. In contrast, forskolin and 3-isobutyl-1 methylxanthine each inhibited migration of unstimulated splenocytes, with little effect on migration of stimulated splenocytes. Only dipyridamole alone, a PDE inhibitor capable of inhibiting PDE8, strongly inhibited migration of stimulated and unstimulated splenocytes and this inhibition was enhanced by forskolin and reversed by a PKA antagonist. Following concanavalin A stimulation, mRNA for PDE8A1 was induced. These results suggest that in employing PDE inhibitor therapy for inflammatory illnesses, inhibition of PDE8 may be required to inhibit migration of activated lymphocytes to achieve a full therapeutic effect. PMID- 16696949 TI - Cloning and expression of maize-leaf pyruvate, Pi dikinase regulatory protein gene. AB - Pyruvate, orthophosphate dikinase (PPDK; E.C. 2.7.9.1) catalyzes the synthesis of the primary inorganic carbon acceptor, phosphoenolpyruvate in the C4 photosynthetic pathway and is reversibly regulated by light. PPDK regulatory protein (RP), a bifunctional serine/threonine kinase-phosphatase, catalyzes both the ADP-dependent inactivation and the Pi-dependent activation of PPDK. Attempts to clone the RP have to date proven unsuccessful. A bioinformatics approach was taken to identify the nucleotide and amino acid sequence of the protein. Based on previously established characteristics including molecular mass, known inter- and intracellular location, functionality, and low level of expression, available databases were interrogated to ultimately identify a single candidate gene. In this paper, we describe the nucleotide and deduced amino acid sequence of this gene and establish its identity as maize PPDK RP by in vitro analysis of its catalytic properties via the cloning and expression of the recombinant protein. PMID- 16696950 TI - Adipogenic differentiation of adipose tissue derived adult stem cells in nude mouse. AB - We evaluated the use of a combination of adipose tissue derived adult stem cells (ADSCs) obtained from liposuction and injectable poly(lactic-co-glycolic acid) (PLGA) spheres for adipose tissue engineering. Adipogenesis was examined in nude mice injected subcutaneously with ADSCs (group I), PLGA spheres (group II), or ADSCs attached PLGA spheres (group III) cultured in adipogenic medium for 7 days. After 4 and 8 weeks, newly formed adipose tissue was observed in groups II and III but not in group I. Oil red O staining of newly formed tissue showed that there was substantially more tissue regeneration and adipogenic differentiation in group III than in group II. RT-PCR confirmed that, after 8 weeks, the PLGA attached ADSCs had fully differentiated into adipocytes. This study provides significant evidence that ADSCs and PLGA spheres can be used in a clinical setting to generate adipose tissue as a noninvasive soft tissue filler. PMID- 16696951 TI - Antagonism of peroxisome proliferator-activated receptor gamma prevents high-fat diet-induced obesity in vivo. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) has been reported to play an important role to regulate adiposity and insulin sensitivity. It is not clear whether antagonism of PPARgamma using a synthetic ligand has significant effects on adipose tissue weight and glucose metabolism in vivo. The aim of this study is to examine the effects of a synthetic PPARgamma antagonist (GW9662) on adiposity and glycemic control in high-fat (HF) diet-fed mice. First the properties of GW9662 as a PPARgamma antagonist were estimated in vitro. GW9662 displaced [(3)H]rosiglitazone from PPARgamma with K(i) values of 13nM, indicating that the affinity of GW9662 for PPARgamma was higher than that of rosiglitazone (110nM). GW9662 had no effect on PPARgamma transactivation in cells expressing human PPARgamma. Treatment of 3T3-L1 preadipocytes with GW9662 did not increase aP2 expression or [(14)C]acetic acid uptake. GW9662 did not recruit transcriptional cofactors to PPARgamma. Limited trypsin digestion of the human PPARgamma/GW9662 complex showed patterns of digestion distinct from those of rosiglitazone. This suggests that the binding characteristics between GW9662 and PPARgamma are different from those of rosiglitazone. Treatment of HF diet-fed mice with GW9662 revealed that this compound prevented HF diet-induced obesity without affecting food intake. GW9662 suppressed any increase in the amount of visceral adipose tissue, but it did not change HF diet-induced glucose intolerance. These data indicate that antagonism of PPARgamma using a synthetic ligand suppresses the increased adiposity observed in HF diet-induced obesity, and that a PPARgamma antagonist could possibly be developed as an anti-obesity drug. PMID- 16696952 TI - Potential output pathways for agonistic-like responses resulting from the GABA(A) blockade of the torus semicircularis dorsalis in weakly electric fish, Gymnotus carapo. AB - The purpose of this study is to examine the pathways involved in the electromotor (electric organ discharge interruptions) and skeletomotor responses (defense like) observed by blockade of GABAergic control of the torus semicircularis dorsalis (TSd) of the awake weakly electric fish Gymnotus carapo, described in a former study. Microinjection of NMDA (5 mM) into the pacemaker nucleus (PM) through a guide cannula previously implanted caused a prolonged interruption of the electric organ discharge (EOD) intermingled with reduction in frequency, similar to that described for TSd GABA(A) blockade, but without noticeable skeletomotor effects. The EOD alterations elicited by bicuculline microinjections (0.245 mM) into the TSd could be blocked or attenuated by a previous microinjection of AP-5 (0.5 mM), an NMDA antagonist, into the PM. Labeled terminals are found in the nucleus electrosensorius (nE) after injection of the biotinylated dextran amine (BDA) tracer into the TSd and into the sublemniscal prepacemaker nucleus (SPPn) subsequent to the tracer injection into the nE. Defense-like responses but not EOD interruptions are observed after microinjections of NMDA (5 mM) into the rhombencephalic reticular formation (RF), where labeled terminals are seen after BDA injection into the TSd and somata are filled after injection of the tracer into the spinal cord. In this last structure, marked fibers are seen subsequent to injection of BDA into the RF. These results suggest that two distinct pathways originate from the torus: one for EOD control, reaching PM through nE and SPPn, and the other one for skeletomotor control reaching premotor reticular neurons. Both paths could be activated by toral GABA(A) blockade. PMID- 16696953 TI - Effects of electroacupuncture on retinal nerve growth factor and brain-derived neurotrophic factor expression in a rat model of retinitis pigmentosa. AB - The aim of this study was to investigate the effect of electroacupuncture (EA) on the progression of retinal degeneration in rats affected by inherited retinitis pigmentosa (IRP) and to correlate this event with the retinal expression of neurotrophins. Thirty-day-old Royal College of Surgeons (RCS) rats were exposed to 25-min-long daily sessions of low-frequency EA for 11 consecutive days. Control-untreated and EA-treated rats were sacrificed 1 h after the last EA session, and their retina removed for biochemical, molecular, and immunohistochemical analyses. Our data revealed that daily sessions of low frequency EA for 11 days to RCS rats during a critical developmental stage of retinal cell degeneration cause an increase of retinal nerve growth factor (NGF) and NGF high-affinity receptor (TrkA) expression; and increase of outer nuclear layer (ONL) thickness; and enhanced vascularization. These findings suggest the possible beneficial effects of EA treatment in the development of IRP-like retinal degeneration of RCS rats and that the mechanism through which EA might exerts its action on the regulation of NGF and brain-derived neurotrophic factor (BDNF) and/or their receptors in retinal cells. PMID- 16696954 TI - Event-related potential correlates of extradimensional and intradimensional set shifts in a modified Wisconsin Card Sorting Test. AB - Event-related potentials (ERPs) were recorded in healthy adult participants during the performance of a modified version of the Wisconsin Card Sorting Test that was designed to isolate the effects of extradimensional (ED) and intradimensional (ID) set-shifts. ERP averages were created for ED- and ID-Shift trials, as well as for the 5th trial in each block (Maintain-Rule). Differences in sensory and longer latency ERP components were found between the ED- and ID conditions, and between the two shift conditions and the Maintain-Rule trials. Consistent with the previous literature, these data indicated that ED- and ID Shifts require different levels of neural resources. A secondary goal of this experiment was to use the excellent temporal resolution of ERPs to examine the neural correlates of various other aspects of the performance of a set-shift task, including differences between correct shifts and the commission of errors, and the differences between the reception of correct and error feedback. Comparisons were made between ERP averages to correct ED-Shift trials and ED Error trials, and to feedback following a correct ED-Shift compared to feedback following an error. As expected, ERP differences were found between correct trials and error trials, and between the ERP correlates of receiving different types of feedback. Overall, these data further indicate the utility of using ERP methodology to study various aspects of complex neuropsychological paradigms. PMID- 16696955 TI - Cerebral asymmetry in 14 year olds born very preterm. AB - The normal pattern of cerebral asymmetry may be altered in neurodevelopmental disorders such as autism and schizophrenia. Babies born very preterm have an increased risk of brain damage, and brain abnormalities which persist into adolescence. This study aimed to ascertain whether preterm birth affects the development of fronto-occipital asymmetry. Structural MRI (magnetic resonance imaging) scans from 14 year old individuals born very preterm (n = 61; mean age 14 years 11 months; 29 male) and age-matched full-term controls (n = 49; mean age 14 years 11 months; 31 male) underwent morphometric analysis, using well validated stereological methods. Measurements of right and left prefrontal, premotor, sensorimotor and occipitoparietal regional volumes were made and asymmetry indices calculated. These factors underwent a reductive factor analysis. There were no significant between-group differences in fronto-occipital asymmetry between the preterm adolescents and their full-term counterparts. It seems unlikely, therefore, that preterm birth per se deviates the development of normal fronto-occipital asymmetry. Neonatal periventricular haemorrhage with ventricular dilatation revealed by ultrasound may be associated with reversal of asymmetry in the sensorimotor area. PMID- 16696956 TI - Peroxisome proliferator-activated receptor-gamma-agonist, rosiglitazone, promotes angiogenesis after focal cerebral ischemia. AB - Peroxisome proliferator-activated receptor-gamma (PPAR-gamma) agonist, rosiglitazone, not only improves insulin resistance in patients with type II diabetes but also exerts a broad spectrum protective effects in variable animal models of neurologic or cardiovascular diseases. We studied the effect of rosiglitazone on angiogenesis and neurological recovery after focal cerebral ischemia. Rosiglitazone (3 mg/kg or 0.3 mg/kg, p.o.) was administered for 7 days prior to and 3 days after the induction of focal ischemia (total 10 days) in adult rats. The rosiglitazone-treated group showed the enhanced neurologic improvement, the reduced infarction volume compared to the ischemia-vehicle group with dose dependency, and the reduced hemispheric atrophy. Rosiglitazone treatment reduced TUNEL(+)/activated caspase-3(+) cells, MPO(+)/Ox-42(+) inflammatory cell infiltrations, caspase-3 activity, and Bax(+) cells, as compared to the ischemia-vehicle group. The vascular surface area, the vascular branch points, the vascular length, and the number of BrdU(+) endothelial cells were significantly increased in the rosiglitazone group compared with the ischemia-vehicle group. Rosiglitazone increased eNOS expression around the ischemic margin with downregulation of FasL. Here, we show that rosiglitazone treatment enhances angiogenesis and functional recovery with dose-dependent induction of ischemic tolerance. PMID- 16696957 TI - Evidence that estrogen directly and indirectly modulates C1 adrenergic bulbospinal neurons in the rostral ventrolateral medulla. AB - Blood pressure in women increases after menopause, and sympathetic tone in female rats decreases with estrogen injections in the rostral ventrolateral medulla (RVLM) region that contains bulbospinal C1 adrenergic neurons and is involved in blood pressure control. We investigated the anatomical and physiological basis for estrogen effects in the RVLM. Neurons with alpha- or beta-subtypes of estrogen receptor (ER) immunoreactivity (-ir) overlapped in distribution with tyrosine hydroxylase (TH)-containing C1 neurons. Immunoelectron microscopy revealed that ERalpha- and ERbeta-ir had distinct cellular and subcellular distributions. ERalpha-ir was most commonly in TH-lacking profiles, many of which were axons and peptide-containing afferents that contacted TH-containing dendrites. ERalpha-ir was also in some TH-containing dendrites. ERbeta-ir was most frequently in TH-containing somata and dendrites, particularly on endoplasmic reticula, mitochondria, and plasma membranes. In whole-cell patch clamp recordings from isolated bulbospinal RVLM neurons, 17beta-estradiol dose dependently reduced voltage-gated Ca(++) currents, especially the long-lasting (L type) component. This inhibition was reversed by washing or prevented by adding the non-subtype-selective ER antagonist ICI182780. An ERbeta-selective agonist, but not an ERalpha-selective agonist, reproduced the Ca(++) current inhibition. The data indicate that estrogens can modulate the function of RVLM C1 bulbospinal neurons either directly, through extranuclear ERbeta, or indirectly through extranuclear ERalpha in selected afferents. Moreover, Ca(++) current inhibition may underlie the decrease in sympathetic tone evoked by local 17beta-estradiol application. These findings provide a structural and functional basis for the effects of estrogens on blood pressure control and suggest a mechanism for the modulation of cardiovascular function by estrogen in women. PMID- 16696958 TI - Estrogen has a neuroprotective effect on axotomized RGCs through ERK signal transduction pathway. AB - The neuroprotective effects of estrogen on neuronal cells in central nervous system have been described previously, however, the mechanisms of neuroprotective effect of estrogen against retinal ganglion cell (RGC) death has not been well identified. To examine the role of endogenous sex steroids produced in ovary, retina samples were prepared from female rats with or without ovariectomy and the density of RGC was calculated. Ovariectomy alone had no effect on the density of fluorogold (FG)-labeled RGC without injury, while the density of surviving RGC after optic nerve axotomy with ovariectomy was significantly decreased compared to that without ovariectomy. To examine the role of exogenous sex steroids, 17beta-estradiol was injected into the vitreous cavity in ovariectomized rats and showed neuroprotective effect on axotomy-induced RGC death while exogenous progesterone showed no effect. Immunoblot and immunohistochemical analysis demonstrated that ERK-c-Fos signal transduction pathway was activated by exogenous 17beta-estradiol in ganglion cell layer. U0126, an ERK inhibitor, inhibited the neuroprotective effect of estrogen on axotomized RGC death. These data suggest that estrogen has neuroprotective effect through activation of ERK-c Fos signaling pathway on axotomy-induced RGC death. The neuroprotective effect of estrogen may have therapeutic benefits in retinal diseases associated with RGC death such as glaucoma. PMID- 16696959 TI - A kinetic study of D-glucose oxidation by bromine in aqueous solutions. AB - The kinetics of the oxidation of D-glucose to D-gluconic acid by bromine in aqueous solution were studied using potentiometric techniques and theoretical considerations of complex bromine-bromide-pH equilibria. The pH has a strong influence on reaction rate. At pH < 8 the reaction is very slow, while in the pH range pH 8-9.5 the reaction is sufficiently fast and seems optimal for the reaction. The proposed active species at that pH region is hypobromous acid. At pH > 9.5, the reaction is further accelerated due to the formation of hypobromite. The proposed kinetics expression for gluconic acid formation, based on the determined kinetic parameters at pH 9.24, is of the form dc(GA)/dt = 160c(2)(G)c(o)(HOBr)c(o)(H(+)c(o)(Br) PMID- 16696960 TI - Photoinduced electron-transfer alpha-deoxygenation of aldonolactones. Efficient synthesis of 2-deoxy-D-arabino-hexono-1,4-lactone. AB - A photoinduced electron-transfer (PET) reaction was used for the deoxygenation at C-2 of aldonolactones derivatized as 2-O-[3-(trifluoromethyl)benzoyl] or benzoyl esters. By irradiation of different D-galactono- and D-glucono-1,4-derivatives, with a 450W lamp, using 9-methylcarbazole as photosensitizer, the corresponding 2 deoxy-D-lyxo- and 2-deoxy-D-arabino-hexono-1,4-lactones were efficiently obtained. PMID- 16696961 TI - Targeting membrane proteins to liquid-ordered phases: molecular self-organization explored by fluorescence correlation spectroscopy. AB - The complex and dynamic architecture of biological membranes comprises of various heterogeneities, some of which may include lipid-based and/or protein-based microdomains called "rafts". Due to interactions among membrane components, several types of domains can form with different characteristics and mechanisms of formation. Model membranes, such as giant unilamellar vesicles (GUVs), provide a key system to study lipid-lipid and lipid-protein interactions, which are potentially relevant to raft formation, by (single-molecule) optical microscopy. Here, we review studies of combined confocal imaging and fluorescence correlation spectroscopy (FCS) on lipid dynamics and organization in domains assembled in GUVs, prepared from various lipid mixtures, which are relevant to the problem of raft formation. Finally, we summarize the results on lipid-protein interactions, which govern the targeting of several putative raft- and non-raft-associated membrane proteins to domain-exhibiting GUVs. PMID- 16696962 TI - Plasma markers of activated hemostasis in the early diagnosis of acute coronary syndromes. AB - BACKGROUND: Because acute coronary syndromes (ACS) are caused by intracoronary thrombosis, plasma markers of coagulation have relevance for early diagnosis. AIMS AND OBJECTIVES: To provide a critical review of these studies and specific attempts to close the diagnostic time gap left by traditional plasma markers of heart injury. METHODS: Studies of ACS patients, with at least one control group, were included when blood samples were taken within 24 h after first symptoms prior to medication or intervention. Special attention was paid to studies reporting diagnostic performance, or combination of several markers into a single diagnostic index. RESULTS: Markers with short plasma half-life (FPA, TAT, etc.) reflect ongoing thrombosis and may identify patients at increased risk. Markers with longer half-life (F1+2, D-Dimer, etc.) may be more useful to indicate a single acute thrombotic event. However, results are highly variable and depend on sampling time, clot property, degree of coronary obstruction and physiological condition. Early diagnostic performance of hemostatic markers was poor even when combined with heart injury markers. CONCLUSIONS: Early measurement of hemostatic plasma markers in ACS patients provides pathophysiological information and may be helpful in risk stratification or to monitor anticoagulant therapy, but does not seem useful in routine clinical diagnosis of ACS. PMID- 16696963 TI - Analytical performance of the Iris iQ200 automated urine microscopy analyzer. AB - BACKGROUND: We evaluated the Iris iQ200 Automated Urine Microscopy Analyzer to find out if the instrument performed better than traditional visual bright field microscopy in detecting basic urine particles, as assessed against reference phase contrast microscopy. METHODS: The HUSLAB quality system was followed in planning and completing the evaluation process. The iQ200 instrument results from 167 mid-stream, uncentrifuged urine specimens were compared to those obtained with phase contrast reference microscopy, and to those with routine bright field microscopy. Linearity, carry-over and precision were tested according to well established protocols. RESULTS: The iQ200 counted erythrocytes (RBC) at r=0.894 (R(2)=0.799) with Automated Particle Recognition (APR) software alone and at r=0.948 (R(2)=0.898) after re-classification. The performance for leukocytes (WBC) was r=0.885 with APR and r=0.978 after re-classification. The correlations of counting after user re-classification were r=0.927 for squamous epithelial cells (SQEP), r=0.856 for casts, and r=0.706 for non-squamous epithelial cells. The iQ200 showed good linearity and precision and no carry-over was detected. CONCLUSIONS: The Iris iQ200 was capable to count reliably RBC, WBC, and SQEP cells and to identify a fraction of bacteria and renal elements. Counting results equalled or exceeded that of routine bright field microscopy or earlier flow cytometric technology. The instrument eliminates manual sample preparation but requires a well-trained technologist for re-grouping of findings. PMID- 16696964 TI - The activity of N-acetyl-beta-D-glucosaminidase and tumor necrosis factor-alpha at early stage of diabetic retinopathy development in type 1 diabetes mellitus children. AB - OBJECTIVE: The assessment of the clinical significance of TNF-alpha, IL-10 and NAG with its A and B isoforms concentrations in children with DM type 1 for the detection of early stages of both diabetic retinopathy and nephropathy. PATIENTS AND METHODS: One hundred and two children with DM type 1 and 35 healthy controls were analyzed. Levels of TNF-alpha, IL10 and total NAG enzyme activity with its A and B isoforms were measured in serum and urine of all participants. RESULTS: Children with diabetic retinopathy had a significantly higher levels of TNFalpha in serum (P=0.01) in comparison to those without retinopathy. The activity of NAG (P=0.002) and its isoform A (P=0.006) and isoform B (P=0.001) were significantly higher in children with diabetic retinopathy in comparison to those without this complication. Conversely, within the group with retinopathy, more children had detectable concentrations of IL10 in serum as compared to those without retinopathy (P=0.01). CONCLUSIONS: These results suggest that NAG activity and TNF-alpha concentration in diabetic retinopathy patients might show a relationship with a degree of renal glomeruli epithelial cells and renal proximal tubules damage. PMID- 16696965 TI - High concentrations of the compatible solute glycinebetaine destabilize model membranes under stress conditions. AB - Compatible solutes are accumulated by diverse organisms in response to environmental stresses such as drought, salt, or cold. Glycinebetaine (Bet) is such a solute that is accumulated by many plants and microorganisms to high concentrations under stress conditions. It is an osmoprotectant in bacteria and stabilizes both soluble and peripherally membrane-bound proteins in vitro. Here, the effects of Bet on the stability of model lipid membranes are compared to the effects of two other compatible solutes, sucrose and trehalose. Both in the presence of 1M NaCl and during freezing to -20 degrees C, Bet is highly destabilizing to liposomes containing nonbilayer lipids, while the disaccharides are either protective or, in some cases, much less destabilizing. The destabilizing effect of Bet is more pronounced in membranes containing the nonbilayer galactolipid monogalactosyldiacylglycerol from plant chloroplasts than in membranes containing the nonbilayer phospholipid phosphatidylethanolamine. The most dramatic differences between the sugars and Bet were observed in liposomes made from a combination of lipids resembling plant chloroplast thylakoid membranes. Measurements with the dye merocyanine 540 indicate that the water membrane interface was affected in opposite directions by the presence of high concentrations of sucrose or Bet. The dynamics of the lipids, however, were not differentially affected by the solutes, making direct solute-lipid interactions an unlikely explanation for the different effects on stability. The data offer an explanation, why Bet at high concentrations achieved during exogenous feeding of leaf tissues can be detrimental to cellular stability and survival under stress, while bacterial membranes that contain phosphatidylethanolamine instead of monogalactosyldiacylglycerol, or cyanobacteria that contain highly saturated monogalactosyldiacylglycerol are less susceptible. PMID- 16696967 TI - Carboxyamido-triazole inhibits proliferation of human breast cancer cells via G(2)/M cell cycle arrest and apoptosis. AB - Carboxyamido-triazole (CAI), a voltage-independent calcium channel inhibitor, has been shown to be able to induce growth inhibition and apoptosis in cancer cells. In the present study, we demonstrate that CAI significantly inhibits proliferation of cultured MCF-7 human breast cancer cells in a dose-dependent manner with an IC(50) of approximately 26 microM. Reduced proliferation of MCF-7 cells in the presence of CAI correlated with accumulation of cells in G(2)/M phase and induction of apoptosis. A treatment of MCF-7 cells with 30 microM CAI caused a time-dependent decrease in the levels of proteins that regulate G(2)/M progression, including Cdk1, Cyclin B1, and Cdc25C. A simultaneous increase in the expression of p21 protein was observed. We also demonstrated a concurrent decrease of the mitochondrial membrane potential (DeltaPsi(m)), and down regulation of anti-apoptotic protein Bcl-2. In conclusion, it seems reasonable to hypothesize that the antitumor effect of CAI in MCF-7 cells is based on G(2)/M cell cycle arrest and inducing apoptosis. PMID- 16696966 TI - Fgf8 expression in the Tbx1 domain causes skeletal abnormalities and modifies the aortic arch but not the outflow tract phenotype of Tbx1 mutants. AB - Fgf8 and Tbx1 have been shown to interact in patterning the aortic arch, and both genes are required in formation and growth of the outflow tract of the heart. However, the nature of the interaction of the two genes is unclear. We have utilized a novel Tbx1(Fgf8) allele which drives Fgf8 expression in Tbx1-positive cells and an inducible Cre-LoxP recombination system to address the role of Fgf8 in Tbx1 positive cells in modulating cardiovascular development. Results support a requirement of Fgf8 in Tbx1 expressing cells to finely control patterning of the aortic arch and great arteries specifically during the pharyngeal arch artery remodeling process and indicate that the endoderm is the most likely site of this interaction. Furthermore, our data suggest that Fgf8 and Tbx1 play independent roles in regulating outflow tract development. This finding is clinically relevant since TBX1 is the candidate for DGS/VCFS, characterized clinically by variable expressivity and reduced penetrance of cardiovascular defects; Fgf8 gene variants may provide molecular clues to this variability. PMID- 16696968 TI - Pyrrolidine dithiocarbamate reduced experimental periodontitis. AB - Nuclear factor-kappaB (NF-kappaB) is a transcription factor which plays a pivotal role in the induction of genes involved in physiological processes as well as in the response to injury and inflammation. Dithiocarbamates are antioxidants which are potent inhibitors of NF-kappaB. We postulated that pyrrolidine dithiocarbamate (PDTC) would attenuate inflammation. In the present study, we have investigated the effects of PDTC, in a rat model of periodontitis. Periodontitis was induced in rats by placing around the lower left first molar a 2/0 braided silk. At day eight the gingivomucosal tissue encircling the mandibular first molar was removed for biochemical and histological analysis. At day eight ligations significantly induced an increase neutrophil infiltration as well as the gingivomucosal tissue expression of TNF-alpha and iNOS as well as nitrotyrosine formation and poly (ADP-ribose) polymerase activation. Ligation significantly increased Evans blue extravasation in gingivomucosal tissue and alveolar bone destruction. Intraperitonial injection of PDTC (10 mg/kg daily for eight days) significantly reduced all of the parameters of inflammation as described above. These data demonstrate that PDTC exerts an anti-inflammatory role during experimental periodontitis and is able to ameliorate the tissue damage associated with ligature-induced periodontitis. PMID- 16696969 TI - AU-rich elements and alternative splicing in the beta-catenin 3'UTR can influence the human beta-catenin mRNA stability. AB - Beta-catenin, the central player of the Wnt signaling cascade, is a well-known oncogene. The regulation of beta-catenin protein stability has been studied extensively while other mechanisms that control cellular levels of beta-catenin have hardly been addressed. In this study, we show that there are three beta catenin mRNA splice variants that differ solely in their 3'-untranslated region (3'UTR) due to alternative splicing or retaining of an intron. The three isoforms were found to be ubiquitously expressed though in different quantities. Upon induction of the beta-catenin protein in peripheral blood mononuclear leukocytes (PBMC), the beta-catenin mRNA is induced in an isoform-specific manner. All three variants occur in the cytoplasm and contribute to the synthesis of beta-catenin acting as a transcriptional coactivator but have different cytoplasmic stabilities in Hela cells. AU-rich elements (AREs), sequence elements implicated in the regulation of mRNA stability, are found in each of the three transcripts. Surprisingly, the AREs contribute to stabilization of the beta-catenin mRNA transcripts in a splicing-dependent manner. The isoform most affected is the one found to be most induced when beta-catenin protein accumulates. These results suggest that alternative splicing and AREs can act together in regulating beta catenin mRNA stability and thereby provide a step of controlling the cellular beta-catenin concentration. PMID- 16696970 TI - Absence of alpha3 (Cx46) and alpha8 (Cx50) connexins leads to cataracts by affecting lens inner fiber cells. AB - Lens development and transparency have been hypothesized to depend on intercellular gap junction channels, consisting of alpha3 (Cx46) and alpha8 (Cx50) connexin subunits, to transport metabolites, secondary messages and ions between lens cells. To evaluate this hypothesis, we have generated alpha3(-/-) alpha8(-/-) double knockout mice and characterized their lens phenotypes. Without gap junctions between lens fiber cells, alpha3(-/-) alpha8(-/-) lenses displayed severe cataracts resulting from cell swelling and degeneration of inner fibers while normal peripheral fiber cells continued to form throughout life. Neither an increase of degraded crystallins nor an increase of water-insoluble crystallins was found in alpha3(-/-) alpha8(-/-) lenses. However, a substantial reduction of gamma-crystallin proteins, but not alpha- and beta-crystallins, was detected. These results suggest that gap junction communication is important for maintaining lens homeostasis of inner fiber cells and that a loss of gap junctions leads to cataract formation as well as reductions of gamma-crystallin proteins and transcripts. PMID- 16696971 TI - Fibroblasts isolated from human pterygia exhibit altered lipid metabolism characteristics. AB - To determine whether the fibrovascular proliferation observed in pterygium, may be, at least in part, mediated by an increased activity of cholesterol metabolism. The correlation between lipid metabolism and rate of growth was studied in human normal conjunctival (NCF) and primary pterygium fibroblasts (PFs) in primary culture. The expression of two proliferation markers (Ki-67 and p53) was evaluated by immunohistochemical staining techniques. Proliferation was evaluated by [(3)H]thymidine incorporation and by immunohistochemical assays. Lipid metabolism was evaluated by (14)C-oleate incorporated into cholesterol esters as well as by oil red O staining. Moreover, the cultures of pterygium fibroblasts were supplemented with two antiproliferative drugs in order to confirm the effective alterations in cholesterol metabolism related to proliferation. Immunohistochemistry of frozen sections from primary pterygium demonstrated an increased staining in Ki-67 and p53 compared with staining observed in normal conjunctiva. A dramatically increased activity of intracellular cholesterol metabolism was demonstrated in pterygium fibroblasts obtained from four different patients. This finding was confirmed by the reduction of cholesterol metabolism in pterygium fibroblasts treated with antiproliferative drugs. Collectively, these data support the hypothesis that alterations of cholesterol metabolism are involved in the development of pterygia. This finding may represent a target of new therapeutic approaches for treatment and prevention of pterygium. PMID- 16696972 TI - High cholesterol content in neurons increases BACE, beta-amyloid, and phosphorylated tau levels in rabbit hippocampus. AB - Epidemiological, cellular, and animal studies suggest that abnormalities in cholesterol metabolism may contribute to the etiology of Alzheimer's disease by increasing the generation of beta-amyloid (Abeta). However, the mechanism by which cholesterol increases Abeta levels is not fully understood. In the present study, we demonstrate that feeding rabbits with 1% cholesterol for 7 months causes an increase in cholesterol content in neurons. High cholesterol content in neurons is accompanied by an increase in the level of BACE1, the enzyme that initially cleaves beta-amyloid precursor protein to generate Abeta, causing the accumulation of Abeta1-42 peptide. These effects correlate with the phosphorylation of tau and the activation of extracellular signal-regulated protein kinase (ERK). Our data suggest that excessive cholesterol content in neurons, following long-term dietary cholesterol, may underlie the increase in BACE1 and Abeta levels. Increased Abeta levels may in turn trigger the phosphorylation of tau by activating ERK. PMID- 16696974 TI - Bone marrow for the brain? PMID- 16696973 TI - Synaptic plasticity in the adult spinal dorsal horn: the appearance of new functional connections following peripheral nerve regeneration. AB - Peripherally regenerated fibers were impaled in the dorsal columns. Each impaled fiber's adequate stimulus was determined and the fiber was activated by passing brief (200 ms) current pulses through the microelectrode. Cord dorsum potentials (CDPs) elicited by fiber stimulation were recorded at 8 sites, and then the fiber was injected with Neurobiotin (NB). In the same preparations, dorsal horn cells were impaled and their receptive fields (RFs) mapped; areas of skin from which the most vigorous responses were elicited were noted. Needle electrodes inserted into these cutaneous "hot spots" were used to electrically activate minimal numbers of peripherally regenerated fibers while simultaneously recording the resulting CDPs and any intracellular EPSPs. This allowed determination of connectivity between regenerated fibers and dorsal horn cells with overlapping RFs. In agreement with findings in intact animals, NB revealed long-ranging collaterals which were not seen using intraaxonally injected horseradish peroxidase (HRP). Although there was no qualitative difference in their morphology compared to those seen in controls, the correlation between spatial distribution of boutons and amplitudes of the monosynaptic CDPs of peripherally regenerated fibers revealed significant shifts in the functional efficacy of many central connections. Transcutaneous electrical stimulation revealed a significantly higher incidence of connectivity between regenerated fibers and cells with overlapping RFs at 9-12 months (86%) than at 5-6 months (34%). Although there was no obvious anatomical reorganization of afferent projections in the dorsal horn, the observed functional changes with time following transection show the formation of new functional central connections. PMID- 16696975 TI - Regulation of histone acetyltransferase activity of p300 and PCAF by proto oncogene protein DEK. AB - The proto-oncogene protein DEK has been implicated in the t(6;9) chromosomal translocation associated with a subtype of acute myelogenous leukemia (AML), which results in the formation of a DEK-CAN fusion protein. Histone acetylation is an important post-translational modification which is involved in transcriptional regulation. In this study, we report that the acidic domain containing protein DEK interacts with histones and exerts a potent inhibitory effect on both p300 and PCAF-mediated histone acetyltransferase activity and transcription. Using chromatin immunoprecipitation assays, we have demonstrated that the recruitment of DEK to the appropriate promoter induces the histone H3 and H4 hypoacetylation of chromatin. Collectively, our data illustrate the important regulatory role played by protein DEK in transcriptional regulation, and suggest that transcription-regulating acidic domain regions may play a role in leukemogenesis. PMID- 16696976 TI - Two mechanistically distinct forms of endocytosis in adrenal chromaffin cells: Differential effects of SH3 domains and amphiphysin antagonism. AB - We previously identified two forms of endocytosis using capacitance measurements in chromaffin cells: rapid endocytosis (RE), dynamin-1 dependent but clathrin independent and slow endocytosis (SE), dynamin-2 and clathrin-dependent. Various recombinant SH3 domains that interact with the proline-rich domain of dynamin were introduced into single cells via the patch pipette. GST-SH3 domains of amphiphysin-1, intersectin-IC, and endophilin-I inhibited SE but had no effect on RE. Grb2-SH3 (N-terminal) or a mutant of amphiphysin-1-SH3 was inactive on either process. These data confirm that dynamin-1 dependent RE is independent of clathrin and show that amphiphysin is exclusively associated with clathrin and dynamin-2-dependent SE. PMID- 16696977 TI - Signaling pathway of ginsenoside-Rg1 leading to nitric oxide production in endothelial cells. AB - We here provide definitive evidence that ginsenoside-Rg1, the pharmacologically active component of ginseng, is a functional ligand of the glucocorticoid receptor (GR) as determined by fluorescence polarization assay. Rg1 increased the phosphorylation of GR, phosphatidylinositol-3 kinase (PI3K), Akt/PKB and endothelial nitric oxide synthase (eNOS) leading to increase nitric oxide (NO) production in human umbilical vein endothelial cell. Rg1-induced eNOS phosphorylation and NO production were significantly reduced by RU486, LY294,002, or SH-6. Also, knockdown of GR completely eliminated the Rg1-induced NO production. This study revealed that Rg1 can indeed serve as an agonist ligand for GR and the activated GR can induce rapid NO production from eNOS via the non transcriptional PI3K/Akt pathway. PMID- 16696978 TI - Critique on the conceptualisation of quality of life: a review and evaluation of different conceptual approaches. AB - Over the past decades, the concept of quality of life has been of paramount importance for evaluating the quality and outcome of health care. Despite its importance, there is still no consensus on the definition or proper measurement of quality of life. Several concept analyses of quality of life have been published. However, they appear to have had a rather limited impact on how empirical studies are conducted. Therefore, we present an overview and critique of different conceptualisations of quality of life, with the ultimate goal of making quality of life a less ambiguous concept. We also describe six conceptual problems. These problems were used as criteria to evaluate the appropriateness of different conceptualisations. This evaluation suggests that defining quality of life in terms of life satisfaction is most appropriate, because this definition successfully deals with all the conceptual problems discussed. The result of our concept evaluation was not surprising for it corroborated the results of several concept analyses and the findings of a structural equation modelling study. Based on the findings revealed by our review, we propose that the scientific community should revitalise the conceptual discussion on quality of life. Furthermore, our findings can assist researchers in developing more rigourous quality-of-life research. PMID- 16696979 TI - The kinetics of Theileria parva infection and lymphocyte transformation in vitro. AB - Theileriaparva is an intracellular protozoan parasite that causes a fatal lymphoproliferative disease of cattle known as East Coast Fever. The parasite infects host lymphocytes causing their transformation and uncontrolled proliferation. Infiltration of major organs with parasitized lymphoblasts results in most cases in death within 3 weeks. Although both T and B lymphocytes are susceptible to infection, the majority of cell lines arising from infection of peripheral blood mononuclear cells in vitro are of T cell lineage. To explore the basis of this phenotypic bias we have followed the very early stages of parasite development in vitro at the single cell level. Peripheral blood mononuclear cells were infected and stained for both surface phenotype and intracellular parasite antigen and analysed by flow cytometry. Although the parasite antigen was detected intracellularly as early as 6h p.i., our data indicate that parasite infection does not lead to cell transformation in all instances. Rather, specific cell types appear to undergo selection very early after infection and expansion of particular cell subsets results in survival and growth of only a small proportion of the cells originally parasitized. PMID- 16696980 TI - Vector-borne parasites decrease host mobility: a field test of freeze or flee behaviour of willow ptarmigan. AB - Transmission mode has been suggested to be a strong predictor of virulence. According to theory, the transmission of vector-borne parasites should be less dependent on host mobility than directly transmitted parasites. This could select for increased exploitation of host resources in parasites transmitted by vectors, which may be manifested as higher virulence. Here, we test the prediction that there is an association between transmission mode and the effect on host mobility by comparing parasite infection levels and mobility in willow ptarmigan (Lagopus lagopus L.). We examined the endoparasite infracommunities of individual hosts to obtain annual, quantitative data on four vector-transmitted species (Leucocytozoon lovati, Trypanosoma avium, Haemoproteus mansoni and microfilaria), two directly transmitted species (Trichostrongylus tenuis and Eimeria sp.) and two species with indirect life cycles (Hymenolepis microps and Parionella urogalli). We then used observed variations in freeze-or-flee responses of individual willow ptarmigan to assess whether parasite intensities were related to scored freezing responses. From a field data set covering a period of 9 years from a single area, we found that stronger freezing responses were associated with higher intensities of vector-borne parasites, especially with higher intensities of the haemosporidian L. lovati. Freezing responses were not associated with parasites transmitted in other ways. Thus, high intensities of vector-borne parasites tended to reduce host movements, while parasites with other transmission modes did not. PMID- 16696981 TI - Dendritic cell activation and function in response to Schistosoma mansoni. AB - Dendritic cells (DC) are uniquely specialised for both antigen acquisition and presentation, linking innate and adaptive immunity. Their central role in the activation of naive T cells gives DC a strategic position in the control of immune responses. While the mechanisms by which viral, bacterial or protozoal pathogens interact with and activate DC are increasingly understood, much less is known about how these cells react to more complex organisms such as schistosomes. Recent studies have examined the impact on DC of antigens from different life cycle stages of Schistosoma mansoni and have revealed a DC phenotype quite distinct to that of conventional activation. Schistosome antigens elicit little of the cytokine secretion and costimulation that are abundantly triggered in DC by unicellular, proinflammatory pathogens and indeed may even actively inhibit such events. The DC response is not a null one, however, since S. mansoni-exposed DC still act as potent antigen presenting cells capable of generating a powerful Th2 immune response. Understanding the interaction between schistosomes and DC is therefore not only addressing fundamental questions of DC biology and immunity to multicellular parasites but also opens the way to therapeutic manipulation of the immune system. PMID- 16696982 TI - Quantitative and qualitative comparisons of Cryptosporidium faecal purification procedures for the isolation of oocysts suitable for proteomic analysis. AB - With the recent publication of the Cryptosporidium genome, investigation of the proteins expressed by Cryptosporidium parvum will provide complementary information on the biology of this complex organism. Proteomic studies on this apicomplexan parasite have been hampered due to the inability to culture or isolate high numbers required for 2D gel analysis. Neonatal calves are a common source of Cryptosporidium oocysts and we report on the development of a sucrose Percoll purification procedure which produced the high yield and purity (free from faecal and bacterial contaminants) that is required for successful proteomic studies from neonatal calves. We report on the development of quantitative and qualitative flow cytometric methods which were confirmed by epifluorescence microscopy. A comparison of five common purification procedures was carried out to determine the efficiency of the sucrose-Percoll gradient. 2D-PAGE results strongly support the sucrose-Percoll procedure as the most suitable method for applications like proteomics which require the recovery of high numbers of isolated oocysts with minimal faecal and bacterial contaminants. PMID- 16696983 TI - Targeting of host cell lineages by vertically transmitted, feminising microsporidia. AB - Feminising microsporidian parasites are transmitted vertically from generation to generation of their crustacean hosts. Little is known about the mechanisms underpinning vertical transmission, in particular, parasite transmission to the host gonad during host development. Here, we investigate the burden and distribution of two species of vertically transmitted, feminising microsporidia (Dictyocoela duebenum and Nosema granulosis) during early embryogenesis (zygote to eight-cells) of the Gammarus duebeni host. Parasite burden differs between the two parasites with N. granulosis being higher by a factor of 10. Whilst D. duebenum replicates during the first few host cell divisions, there is no increase in N. granulosis burden. Only merogonic parasite stages were observed in the host embryo. Distribution of both parasites was non-random from the two-cell embryo stage, indicating biased parasite segregation at host cell division. Dictyocoela duebenum burden was low in the germline and somatic gonad progenitor cells but was highest in the ectoderm precursors, leading us to propose that the parasite targets these cells and then secondarily infects the gonad later in host development. Targeting by N. granulosis was less specific although there was a persistent bias in parasite distribution throughout host cell divisions. Parasite burden was highest in the ectoderm precursors as well as the germline progenitors leading us to suggest that, in addition to using the ectodermal route, N. granulosis may also target germline directly. Biased segregation will be adaptive for these parasites as it is likely to lead to efficient transmission and feminisation whilst minimising virulence in the host. PMID- 16696984 TI - Coronary collateral circulation: the effects of smoking and alcohol. AB - OBJECTIVE: The presence or absence of coronary collaterals is of vital importance during acute ischemia. Smoking and alcohol have been suggested to play a role, but data are scarce. We examined the extent to which smoking and alcohol use affect the presence of coronary collateral circulation. METHODS: Cross-sectional study in 242 patients, admitted for elective PTCA. Smoking was defined as past or current. Pack years were calculated and categorized into never-smokers (reference category): <10, 10-19, 20-29, and >or=30 pack years. Alcohol consumption was defined as past or current, and categorized into never-users (reference category): <1, 1-10, 11-20, and >or=21 units per week (UPW). Collaterals were graded with Rentrop's classification. Coronary collateral presence was defined as Rentrop-grade >or=1. RESULTS: Current smoking (odds ratio (OR) 4.17; 95% confidence interval (CI) 1.79-9.71) was positively associated, while pack years of smoking was not related. Current alcohol intake showed a J-shaped tendency with coronary collateral presence, while past moderate alcohol consumption was inversely associated (OR 0.19; 95% CI 0.04-0.98). CONCLUSIONS: Smoking and (to some extent) alcohol use are associated with collateral presence. The results support the view that life-style factors may affect the formation of coronary collaterals in patients with ischemic cardiac disease. PMID- 16696985 TI - Different elution modes and field programming in gravitational field-flow fractionation: field programming using density and viscosity gradients. AB - In previous papers, several approaches to programming of the resulting force field in GFFF were described and investigated. The experiments were dealing with flow-velocity and channel thickness, i.e. factors influencing hydrodynamic lift forces (HLF). The potential of density and viscosity of carrier liquid for field programming was predicted and demonstrated by preliminary experiments. This work is devoted to experimental verification of the influence of carrier liquid density and viscosity. Several carrier liquid density and simultaneously viscosity gradients using water-methanol mixtures are in this work implemented in the separation of a model silica mixture. Working with the water-methanol gradients, one is not able to separate the influence of density from the contribution of viscosity. However, we found experimental conditions to show the isolated effect of carrier liquid density (two water-methanol mixtures of equal viscosity differing in their densities). In order to demonstrate the isolated effect of viscosity, we implemented in this work a new system of (hydroxypropyl)methyl cellulose (HPMC) carrier liquids. Three different HPMC compositions enabled to vary the viscosity more than two times at almost constant density. With increasing carrier liquid viscosity, the focusing and elevating trend was clearly pronounced for 5 and 10 microm silica particles. By the isolated effect of increased viscosity, the centre of the 10 microm particle zone was elevated to the streamline at 16% of the channel height. These experiments have shown that the influence of carrier liquid viscosity on HLF should be taken into account even at higher levels above the channel bottom, i.e. beyond the near wall region. Further, it is shown that higher value of carrier liquid viscosity improves the separation of the model mixture in terms of time and resolution. PMID- 16696986 TI - Electrokinetic migration across artificial liquid membranes Tuning the membrane chemistry to different types of drug substances. AB - Twenty different basic drugs were electrokinetically extracted across a thin artificial organic liquid membrane with a 300 V d.c. electrical potential difference as the driving force. From a 300 microl aqueous sample (acidified corresponding to 10mM HCl), the drugs were extracted for 5 min through a 200 microm artificial liquid membrane of a water immiscible organic solvent immobilized in the pores of a polypropylene hollow fiber, and into a 30 microl aqueous acceptor solution of 10mM HCl inside the lumen of the hollow fiber. Hydrophobic basic drugs (logP>1.7) were effectively isolated utilizing 2 nitrophenyl octyl ether (NPOE) as the artificial liquid membrane, with recoveries up to 83%. For more hydrophilic basic drugs (logP<1.0), a mixture of NPOE and 25% (w/w) di-(2-ethylhexyl) phosphate (DEHP) was required to ensure efficient extraction, resulting in recoveries up to 75%. DEHP was expected to act as an ion pair reagent ion-pairing the protonated hydrophilic drugs at the interface between the sample and the membrane, resulting in permeation of the interface. PMID- 16696987 TI - Simultaneous analysis of Vitamins A and E in infant milk-based formulae by normal phase high-performance liquid chromatography-diode array detection using a short narrow-bore column. AB - A rapid, simple and reproducible normal-phase (NP) high-performance liquid chromatography (HPLC)-diode array detection (DAD) method for simultaneous qualitative and quantitative determination of Vitamin A (retinol acetate and retinol palmitate) and Vitamin E (alpha-tocopherol acetate, alpha-, gamma- and delta-tocopherols) in milk-based infant formulae was developed and validated. The preparation sample was based on protein precipitation and vitamin extraction with ethanol, followed by re-extraction with hexane, while the chromatographic method was based on the use of a short narrow-bore column (50 mm x 2.1 mm; 3 microm particle size), which afforded less solvent consumption and higher mass sensitivity. The method showed acceptable values for precision, recovery and sensitivity, and proved very simple for routine analysis work. PMID- 16696988 TI - Simplified analysis of organic compounds in headspace and aqueous samples by high capacity sample enrichment probe. AB - A sample enrichment probe (SEP) consisting of a thin rod of an inert material and provided at one end with a short sleeve of polydimethylsilicone rubber was used for the high-capacity sample enrichment of analytes from gaseous and aqueous samples for analysis by gas chromatography (GC) and its hyphenated techniques. The silicone rubber was exposed to the analytical sample, after which the end of the rod carrying the silicone rubber was introduced into the injector and the analytes thermally desorbed and analysed by GC. This technique is similar to, but differs from, solid-phase microextraction (SPME) in that a much larger volume of the sorptive phase is employed, the sorptive phase is not introduced into the inlet of the GC via a needle and the injector is opened to the atmosphere for the introduction and removal of the SEP. In the determination of volatile and semi volatile organic compounds in gaseous and aqueous media, the SEP technique gave results comparable with those obtained by the stir-bar-sorptive extraction (SBSE) and high-capacity sorption probe (HCSP) techniques. Implementation of the SEP technique requires only minor adaptations to the gas chromatograph and does not require any auxiliary thermal desorption and cryotrapping equipment. PMID- 16696989 TI - Identification of three novel compounds in wine by means of a laboratory constructed multidimensional gas chromatographic system. AB - A laboratory-made off-line multidimensional GC-GC system has been optimized and applied to the isolation and identification of three novel aroma compounds in wine. The system comprises two independent chromatographs equipped with four-port manually actuated valves and fast capillary connectors that make it possible to quickly transfer a capillary trapping loop from the first to the second system. The first system includes a split/splitless injector, a thick phase semi capillary column, the valve and both a flame ionization detection (FID) system and a sniffing port. The trapping loop lies outside the chromatograph and it is immersed in a Dewar with liquid nitrogen. The second system includes a programmable temperature vaporization (PTV) injector, the valve, the analytical column and two parallel detectors (a MS system and a sniffing port). The injection of large sample volumes (90 microl) in the first GC system and the general operation of the system have been optimized. The extract was obtained by purging the wine at 37 degrees C with a stream of nitrogen and trapping the volatiles in a bed containing 1g LiChrolut EN. Fatty acids and alcohols were removed by washing the resins with a water/methanol solution containing 1% NaHCO3, and volatiles were recovered with dichloromethane. The GC-MS analysis of the cuts obtained in the first GC system made it possible to identify by first time in wine the strong smelling compounds ethyl 2-, 3- and 4-methylpentanoate. PMID- 16696990 TI - Development and application of microporous hollow fiber protected liquid-phase microextraction via gaseous diffusion to the determination of phenols in water. AB - A new organic solvent-free microextraction technique termed liquid-gas-liquid microextraction (LGLME) was developed. In this technique, a small amount (6 microl) of aqueous acceptor solution (0.5M NaOH) is introduced into the channel of a 2.65 cm polypropylene hollow fiber. The hollow fiber is then immersed in an aqueous sample donor solution. The aqueous acceptor phase in the channel of the hollow fiber is separated from the sample solution by the hydrophobic microporous hollow fiber wall with air inside its pores. The analytes (phenols) passed through the microporous hollow fiber membrane by gas diffusion and were then trapped by the basic acceptor solution. After extraction, the acceptor solution was withdrawn into a microsyringe and injected into a capillary electrophoresis sample vial for subsequent analysis. Limits of detection of between 0.5 and 10 microg/l for eight phenols could be achieved. The relative standard deviations (n=6) of this technique between 2.7 and 7.6%. The technique also provides good enrichment factors for all the eight analytes. PMID- 16696991 TI - New stationary phases for high-performance liquid chromatography based on poly(methyltetradecylsiloxane) thermally immobilized onto zirconized silica. AB - The modification of silica with zirconium oxide followed by sorption and thermal immobilization of poly(methyltetradecylsiloxane) (PMTDS) is used to prepare a reversed stationary phase for high-performance liquid chromatography. The thermal immobilization of PMTDS on zirconized silica was optimized using a central composite design. The new stationary phase was characterized by spectroscopic and chromatographic methods. Stationary phases with good reproducibility and good chromatographic performance for various compounds were obtained. PMTDS thermally immobilized on zirconized silica presented quite significant chemical stability at pH 10 and 50 degrees C. PMID- 16696992 TI - Enhancing micro-scale membrane extraction by implementing a barrier film. AB - Hollow fiber based microextraction techniques have shown much promise in the extraction and preconcentration of analytes. Typically, a solvent held in the membrane is used as an extractant (or an acceptor). However, the acceptor also permeates out during long extractions and vigorous agitation. The stabilization of the acceptor is important for high enrichment and good reproducibility. Coating the membrane with a low permeability barrier film can provide diffusional resistance to reduce the outflow of the acceptor. Several compounds were evaluated as potential barrier films. They were found to protect the organic acceptor and significantly enhance extraction performance. In a study with six polycyclic aromatic hydrocarbons (PAHs), enrichment factors ranged between 494 and 4555 were obtained and the detection limits were at low ppb to ppt levels. Good reproducibility with RSDs between 1.60 and 7.65%, and large linear dynamic ranges with good linearity (R2 between 0.9870 and 0.9997) were also achieved. PMID- 16696993 TI - New method for determination of ochratoxin A in beer using zinc acetate and solid phase extraction silica cartridges. AB - A new method for the determination of ochratoxin A (OTA) in beer has been developed. The new method has been compared with a reference method currently accepted as AOAC official first action. The limits of detection and quantification of the proposed method were 0.0008 and 0.0025 ng/ml, respectively, while they were 0.0025 and 0.0075 ng/ml, respectively, in the AOAC method used as reference. The recovery levels in the 0.025-0.40 ng OTA/ml spiking range for the proposed and the reference methods were 80.6-87.6% and 78.2-83.8%, respectively. The relative standard deviations of recoveries were 2.6-7.5% for the proposed method and 0.7-6.1% for the reference method. Passing and Bablok regression analysis of recovery data obtained by the proposed method versus data obtained by the reference method on an OTA-spiked beer sample showed good correlation (r2 = 0.9993), while the slope and intercept were 1.049 and -0.0013, respectively. The advantage of the proposed method is the low cost of the materials used in sample preparation because expensive immunoaffinity columns are not needed to clean-up samples while it maintains or even increases the good performance of the reference method. The proposed method was applied to 69 beer samples from different geographic origins (national and imported) but purchased in the Spanish market. They were found to be contaminated with OTA in the range from 0.008 to 0.498 ng/ml (average: 0.070 ng/ml). Five samples surpassed the limit recommended by the European Union (0.2 ng OTA/g). PMID- 16696994 TI - Growth of ultrathin films of cadmium telluride and tellurium as studied by electrochemical atomic force microscopy. AB - Time dependent, cathodic electrodeposition of ultrathin CdTe and Te films has been studied in 50 mM H(2)SO(4) + 1 mM CdSO(4) + 0.1 mM TeO(2) solutions at room temperature under potential control using electrochemical atomic force microscopy (EC-AFM). The films were also characterized electrochemically and with X-ray diffraction. The growth mechanism and the composition of the films depends on the applied potentials. Island-like growth mode was observed for CdTe films when the deposition potential was -0.35 V (SHE). At a more positive deposition potential of 0.138 V (SHE), Cd was not co-deposited into the film but affected the dynamic growth mode of the deposit. At this voltage smooth Te films were obtained. Depending on the applied potential, Cd acts either as a co-deposition element for CdTe film growth, or as a mediator for layer-by-layer growth of Te films. PMID- 16696995 TI - Removal of bromide and iodide anions from drinking water by silver-activated carbon aerogels. AB - The aim of this study is to analyze the use of Ag-doped activated carbon aerogels for bromide and iodide removal from drinking water and to study how the activation of Ag-doped aerogels affects their behavior. It has been observed that the carbonization treatment and activation process of Ag-doped aerogels increased the surface area value ( [Formula: see text] ), whereas the volume of meso-(V(2)) and macropores (V(3)) decreased slightly. Chemical characterization of the materials revealed that carbonization and especially activation process considerably increased the surface basicity of the sample. Original sample (A) presented acidic surface properties (pH(PZC)=4.5) with 21% surface oxygen, whereas the sample that underwent activation showed mainly basic surface chemical properties (pH(PZC)=9.5) with only 6% of surface oxygen. Carbonization and especially, activation process considerable increased the adsorption capacity of bromide and iodide ions. This would mainly be produced by (i) an increase in the microporosity of the sample, which increases Ag-adsorption sites available to halide anions, and (ii) a rise of the basicity of the sample, which produces an increase in attractive electrostatic interactions between the aerogel surface, positively charged at the working pH (pH(solution) T transitions are extensive. However, when the break region is not A, T-rich, mutations are mainly restricted to the break, similar to findings in vivo. The results indicate that AID has activity on double strand breaks (DSBs). Based on previous and current findings, a somatic hypermutation (SHM) model is proposed, in which collision between the transcription apparatus and the replication fork generates DSBs. After AID acts on break ends, the error-prone DNA repair machinery fixes and creates mutations. PMID- 16697046 TI - Selective dopamine D3 receptor antagonists enhance cortical acetylcholine levels measured with high-performance liquid chromatography/tandem mass spectrometry without anti-cholinesterases. AB - The present study compared the effects of two selective dopamine (DA) D(3) receptor antagonists, SB-277011A (3, 10 and 30 mg/kg i.p.) and SB-414796A (3, 10 and 30 mg/kg i.p.) on extracellular levels of acetylcholine (ACh) in the rat medial prefrontal cortex (mPFC) by using a LC/MS-MS analytical method that permitted the detection of ACh without the necessity of adding acetylcholinesterase inhibitors to the perfusate. Furthermore, the present LC/MS MS method permitted the simultaneous measurement of the respective concentrations of SB-277011A and SB-414796A in the same extracellular samples from the mPFC. The systemic administration of both selective DA D(3) receptor antagonists produced a significant increase in extracellular levels of Ach compared to vehicle-treated animals, which was associated with increases in extracellular concentrations of SB-277011A and SB-414796. Overall, the present findings further strengthen the likelihood of a modulation of cortical cholinergic function through a DA D(3) mediated mechanism and suggest that selective DA D(3) receptor antagonism may be beneficial in the treatment of psychiatric diseases, such as schizophrenia, which are characterized by cognitive dysfunction. PMID- 16697047 TI - The effects of a celebrity suicide on suicide rates in Hong Kong. AB - BACKGROUND: Deaths of celebrities, especially by suicide, can be followed by an increase in population suicide rates, particularly where there is extensive media reporting. We have examined the impact on suicides following the death of a famous Hong Kong pop singer whose death from suicide by jumping from a height, occurred on 1st April 2003, and resulted in extensive and often dramatic media coverage. METHODS: Data on suicides were obtained from the Hong Kong Census and Statistics Department and the Coroner's Court. The numbers of suicides in 2003 before and after the death of celebrity were compared to the same period in 1998 2002. The case files and suicide notes of people who died by suicide in 2003 were also studied qualitatively. RESULTS: There was a significant increase in suicides following the celebrity death, compared with the average over the preceding three months as well as the corresponding monthly average during 1998-2002. It was particularly marked in a subgroup comprising males, aged 25-39 years, many of whom died by jumping. The name of the celebrity was often mentioned in case files and suicide notes. LIMITATIONS: The statistical results in showing the excess of suicides were based on aggregated data only. CONCLUSIONS: This study provides further confirmation of the potential harmful consequences of sensational and excessive reporting of celebrity deaths. PMID- 16697048 TI - Effect of hedonic tone on event-related potential measures of cognitive processing. AB - Anhedonia, the inability to experience pleasure, is a core feature of several psychiatric disorders such as schizophrenia, depression, and substance dependence. Furthermore, it has been suggested that anhedonia is an important predictor of schizophrenia. Anhedonia has been associated with information processing deficits, especially attentional deficits, which may predispose for schizophrenia. In the present study, event-related potentials (ERPs) were used to examine the influence of hedonic tone on information-processing characteristics in a sample of healthy individuals. Thirty-five healthy subjects were divided into two groups based on their scores on the Snaith-Hamilton Pleasure Scale (SHAPS). Cognitive functions were measured during an active visual oddball paradigm. It was found that both early, middle and late ERP components of subjects with low levels of hedonic tone were attenuated compared with ERPs of subjects with high levels of hedonic tone. These findings suggest that decreased hedonic tone is associated with reductions in both automatic and effortful cognitive processing of relevant stimuli. Consequences of these findings for the vulnerability to psychopathology are discussed. PMID- 16697049 TI - Demonstration of high prevalence of SS-A antibodies in a general population: association with HLA-DR and enterovirus antibodies. AB - Autoantibodies are helpful markers for diagnosing autoimmune diseases and there is a link between HLA-DR3 and the prevalence of SS-A antibodies in clinical groups. We aimed to study this association at the level of general adult population and to verify whether these antibodies are more common in persons with antibodies against enteroviruses as possible associates of Sjogren syndrome. The studied material included sera from 200 persons, randomly selected from a general population sample. The IgG type of SS-A/SS-B autoantibodies were measured by nuclear immunoblot, developed by us, and the results were compared to other results obtained by anti-SS-A immunoblot and ELISA. Enterovirus antibodies were detected by ELISA using common enterovirus antigenic peptide KEVPALTAVETGAT. Altogether 33 out of 200 sera showed SS-A and/or SS-B bands in immunoblot, including all seven ANA Profile 3 (Euroimmune) positive sera. One of the persons positive in these two tests showed also positive reaction on anti-SS-A ELISA (Euroimmune). None of the antibody-positive persons had Sjogren's syndrome or other rheumatic disease. Among 82 HLA typed persons, selected at random, the HLA DRB1*03 and HLA-DRB1*11 allele carriers included significantly more persons with SS-A antibodies than the non-carries (p = 0.008). Antibodies against enterovirus peptide were present more frequently in persons with SS-A autoantibodies than in age- and sex-matched controls (p = 0.009). Summing up, our study showed that the prevalence of SS-A/SS-B antibodies in a general random population might be higher than thought previously being detected in up to 16.5% of persons including a significant number of those with HLA-DR3 or/and DR11 alleles and with antibodies against enteroviruses. Whether all these persons have the risk of developing rheumatic diseases should be evaluated by follow up studies. PMID- 16697050 TI - CSF cytokine and chemokine profiles in acute disseminated encephalomyelitis. AB - We simultaneously measured 16 cytokines/chemokines in cerebrospinal fluid (CSF) from 14 patients with acute disseminated encephalomyelitis (ADEM) and 20 controls using a fluorescent bead-based immunoassay. A variety of cytokines, such as IL 1beta, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IFN-gamma, TNF-alpha, G-CSF and MIP 1beta, were significantly elevated in ADEM. In particular, G-CSF showed a marked 38-fold increase compared to the control mean. Significant positive correlations with inflammatory parameters in CSF, such as cell counts and protein levels, were found for IFN-gamma, IL-6 and IL-8. In contrast, IL-17 produced by activated CD4(+) memory T cells was not increased. The results suggested that various cytokines related to activation of macrophages/microglias and Th(1) and Th(2) cells are upregulated in CSF in ADEM. PMID- 16697051 TI - Myasthenia gravis induced in mice by immunization with the recombinant extracellular domain of rat muscle-specific kinase (MuSK). AB - Myasthenia gravis (MG) is mostly caused by anti-acetylcholine receptor (AChR) auto-antibodies (Abs). Such Abs are undetectable in 10-15% of MG patients, but many have anti-muscle-specific kinase (MuSK) Abs. We injected recombinant rat MuSK extracellular domain in H-2(a), H-2(b), H-2(bm12) and H-2(d) mice. Certain strains exhibited exercise-induced fatigue, tremors, weight loss, and some died after 2-3 injections. Compound muscle action potentials showed decrement with low frequency repetitive nerve stimulation. Miniature endplate potentials decreased, suggesting lower numbers of endplates functional AChRs. Myasthenic sera inhibited agrin-induced AChR aggregation in C2C12 myotubes. CONCLUSION: Anti-MuSK Abs induce MG, which might also result from blocking the agrin-signaling pathway. PMID- 16697052 TI - Aluminum and copper in drinking water enhance inflammatory or oxidative events specifically in the brain. AB - Inflammatory and oxidative events are up-regulated in the brain of AD patients. It has been reported that in animal models of AD, exposure to aluminum (Al) or copper (Cu) enhanced oxidative events and accumulation of amyloid beta (Abeta) peptides. The present study was designed to evaluate the effect of a 3-month exposure of mice to copper sulfate (8 microM), aluminum lactate (10 or 100 microM), or a combination of the salts. Results suggest that although Al or Cu may independently initiate inflammatory or oxidative events, they may function cooperatively to increase APP levels. PMID- 16697053 TI - Innate immune response gene expression profiles of N9 microglia are pathogen-type specific. AB - Glial cells, particularly microglia, are thought to play a pivotal role in initiating and guiding innate immune responses to CNS infections and in perpetuating inflammation and pathology in CNS diseases such as multiple sclerosis and Alzheimer's disease. We describe here the development and use of a new microarray designed to specifically profile transcript expression of innate immunity genes. Microarray analysis validated by quantitative PCR demonstrated an extensive range of pattern recognition receptor gene expression in resting N9 microglia, including Toll-like receptors, scavenger receptors and lectins. Stimulation with LPS or infection with virus modulated pattern recognition receptor, cytokine, chemokine and other innate immune transcripts in a distinct and stimulus-specific manner. This study demonstrates that a single glial cell phenotype has an innate capability to detect infection, determine its form and generate specific responses. PMID- 16697054 TI - Emerging cancer therapeutic opportunities target DNA-repair systems. AB - DNA-damaging agents have a central role in non-surgical cancer treatment. The balance between DNA damage and repair determines the final therapeutic consequences. An elevated DNA-repair capacity in tumor cells leads to drug or radiation resistance and severely limits the efficacy of these agents. Interference with DNA repair has emerged as an important approach in combination therapy against cancer. Anticancer targets in DNA-repair systems have emerged, against which several small-molecule compounds are currently undergoing clinical trials. PMID- 16697055 TI - Subtype-specific alpha1- and beta-adrenoceptor signaling in the heart. AB - Recent studies of adrenoceptors have revealed subtype-specific signaling, promiscuous G-protein coupling, time-dependent switching of intracellular signaling pathways, intermolecular interactions within or between adrenoceptor subfamilies, and G-protein-independent signaling pathways. These findings have extended the classical linear paradigm of G-protein-coupled receptor signaling to a complex "signalome" in which an individual adrenoceptor initiates multiple signaling pathways in a temporally and spatially regulated manner. In particular, persistent stimulation of beta-adrenoceptor subtypes causes a time-dependent switch of signaling pathways and elicits different, even opposing, functional roles of these receptors in regulating cardiac structure and function. Recent progress in the understanding of subtype-specific functions and signaling mechanisms of cardiac adrenoceptor subtypes, particularly beta(1)-adrenoceptors, beta(2)-adrenoceptors, alpha(1A)-adrenoceptors and alpha(1B)-adrenoceptors, might have important pathogenic and therapeutic implications for heart disease. PMID- 16697056 TI - The ClC-3 Cl- channel in cell volume regulation, proliferation and apoptosis in vascular smooth muscle cells. AB - The volume-regulated Cl(-) current (I(Cl.vol)) is responsible for the transmembrane Cl(-) transport that is involved in cell volume regulatory mechanisms. Although the regulation of cell volume is a fundamental function of healthy cells for maintaining constant size, the molecular genetic identification of I(Cl.vol) is still being debated. Recent studies in vascular smooth muscle support the idea that ClC-3, a member of the voltage-gated ClC Cl(-) channel family, is the molecular component involved in the activation or regulation of I(Cl.vol). Moreover, gene-targeting studies in vascular smooth muscle cells (VSMCs) and other cell types indicate emerging roles of ClC-3 in cell proliferation and apoptosis. These findings indicate that ClC-3 might be involved in modulating vascular remodeling in hypertension and arteriosclerosis. PMID- 16697057 TI - Plasticity and specificity of cortical processing networks. AB - The cerebral cortex is subdivided into discrete functional areas that are defined by specific properties, including the presence of different cell types, molecular expression patterns, microcircuitry and long-range connectivity. These properties enable different areas of cortex to carry out distinct functions. Emerging data argue that the particular structure and identity of cortical areas derives not only from specific inputs but also from unique processing networks. The aim of this review is to summarize current information on the interplay of intrinsic molecular cues with activity patterns that are driven by sensory experience and shape cortical networks as they develop, emphasizing synaptic connections in networks that process vision. This review is part of the TINS special issue on The Neural Substrates of Cognition. PMID- 16697058 TI - Feature-based attention in visual cortex. AB - Although most studies of visual attention have examined the effects of shifting attention between different locations in the visual field, attention can also be directed to particular visual features, such as a color, orientation or a direction of motion. Single-unit studies have shown that attention to a feature modulates neuronal signals in a range of areas in monkey visual cortex. The location-independent property of feature-based attention makes it particularly well suited to modify selectively the neural representations of stimuli or parts within complex visual scenes that match the currently attended feature. This review is part of the TINS special issue on The Neural Substrates of Cognition. PMID- 16697059 TI - Hippocampal lesions impair performance on a conditional delayed matching and non matching to position task in the rat. AB - The hippocampus is thought to be involved in a range of cognitive processes, from the ability to acquire new memories, to the ability to learn about spatial relationships. Humans and monkeys with damage to the hippocampus are typically impaired on delayed matching to sample tasks, of which the operant delayed matching to position task (DMTP) is a rat analogue. The reported effects of hippocampal damage on DMTP vary, ranging from delay-dependent deficits to no deficit whatsoever. The present study investigates a novel memory task; the conditional delayed matching/non-matching to position task (CDM/NMTP) in the Skinner box. CDM/NMTP uses the presence of specific stimulus cues to signify whether a particular trial is matching or non-matching in nature. Thus, it incorporates both the task contingencies within one session, and supplements the requirement for remembering the side of the lever in the sample phase with attending to the stimulus and remembering the conditional discrimination for the rule. Rats were trained preoperatively and the effects of bilateral excitotoxic lesions of the hippocampus were examined on postoperative retention of the task. Rats with lesions of the hippocampus incurred a significant impairment on the task that was manifest at all delays intervals. Despite a bias towards matching during training, trials of either type were performed with equivalent accuracy and neither rule was affected differentially by the lesion. This task may prove useful in determining the cognitive roles of a range of brain areas. PMID- 16697060 TI - Behavioural effects of chronic haloperidol and risperidone treatment in rats. AB - The therapeutic properties of typical antipsychotic drugs (APDs) such as haloperidol in schizophrenia treatment are mainly associated with their ability to block dopamine D2 receptors. This blockade is accompanied by side effects such as extrapyramidal symptoms (EPS). Atypical APDs such as risperidone have superior therapeutic efficacy possibly due to their activity at multiple receptors (in particular 5-HT2A receptors). Although the risk of EPS is significantly lower in atypical than in typical APDs, it is not negligible. To investigate and compare the behavioural profile and EPS-asssociated side effects of haloperidol and risperidone APD treatment we applied a multi-tiered, comprehensive behavioural phenotyping approach. Sprague-Dawley rats were treated chronically (28 days) with supratherapeutic EPS-inducing doses of haloperidol and risperidone using osmotic minipumps. Domains such as motor activity, exploration, memory, and anxiety were analysed together with EPS assessment ("early onset" vacuous chewing movements and catalepsy). Both APDs produced diminished motor activity and exploration, impaired working memory performances, and increased anxiety levels. These effects were more pronounced in haloperidol-treated animals. Chronic APD treatment also caused a time-course dependent elevation of EPS-like symptoms. Risperidone treated animals showed a catalepsy-like phenotype, which differed to that of haloperidol-treated rats, indicating that processes other than the anticipated dopaminergic mechanisms are underlying this phenomenon. These EPS-related phenotypes are consistent with reported EPS-inducing D2 receptor occupancies of around 80%. Differences in the behavioural profile of haloperidol and risperidone, which were revealed by a comprehensive phenotyping strategy, are likely due to the unique receptor activation profiles of these APDs. PMID- 16697061 TI - A ubiquitous Plasmodium protein displays a unique surface labeling pattern in sporozoites. AB - The Plasmodium sporozoite is infective for mosquito salivary glands and vertebrate host tissues. Although it is a key developmental stage of the malaria parasite, relatively few sporozoite surface or secreted proteins have been identified and characterized. Herein, we describe the molecular and cellular characterization of a novel surface molecule that is preferentially-expressed in salivary gland sporozoites as compared to oocyst and hemolymph sporozoites. This molecule, designated the sporozoite and erythrocytic stages (SES) protein (formerly known as Pg4), exhibits a spiral surface labeling pattern that spans over a known sporozoite surface antigen, the circumsporozoite protein, with only minor co-localization. SES consists of 551 amino acids encoding a putative 63.2kDa protein that has been shown to be expressed not only on particular sporozoite stages, but also during the asexual and gametocyte stages. This novel protein also has three domains of unknown function that are conserved in at least eight Plasmodium spp. that represent human, avian, non-human primate, and rodent malarias. PMID- 16697062 TI - Iron-dependent metabolic remodeling in S. cerevisiae. AB - All eukaryotes require iron although iron is not readily bioavailable. Organisms expend much effort in acquiring iron and in response have evolved multiple mechanisms to acquire iron. Because iron is essential, organisms prioritize the iron use when iron is limiting; iron-sparing enzymes or metabolic pathways are utilized at the expense of iron-rich enzymes. A large percentage of cellular iron containing proteins is devoted to oxygen binding or metabolism, therefore, changes in oxygen availability affect iron usage. Transcriptional and post transcriptional mechanisms have been shown to affect the concentration of iron containing proteins under iron or oxygen limiting conditions. In this review, we describe how the budding yeast Saccharomyces cerevisiae utilizes multiple mechanisms to optimize iron usage under iron limiting conditions. PMID- 16697063 TI - Trk receptor binding and neurotrophin/fibroblast growth factor (FGF)-dependent activation of the FGF receptor substrate (FRS)-3. AB - We have investigated the signaling properties of the fibroblast growth factor (FGF) receptor substrate 3 (FRS3), also known as SNT-2 or FRS2beta, in neurotrophin-dependent differentiation in comparison with the related adapter FRS2 (SNT1 or FRS2alpha). We demonstrate that FRS3 binds all neurotrophin Trk receptor tyrosine kinases and becomes tyrosine phosphorylated in response to NGF, BDNF, NT-3 and FGF stimulation in transfected cells and/or primary cortical neurons. Second, the signaling molecules Grb2 and Shp2 bind FRS3 at consensus sites that are highly conserved among FRS family members and that Shp2, in turn, becomes tyrosine phosphorylated. While FRS3 over-expression in PC12 cells neither increases NGF-induced neuritogenesis nor activation of Map kinase/AKT, comparable to previous reports on FRS2, over-expression of a chimeric adapter containing the PH/PTB domains of the insulin receptor substrate (IRS) 2, in place of the PTB domain of FRS3 (IRS2-FRS3) supports insulin-dependent Map kinase activation and neurite outgrowth in PC12 cells. Collectively, these data demonstrate that FRS3 supports ligand-induced Map kinase activation and that the chimeric IRS2-FRS3 adapter is stimulating sufficient levels of activated MapK to support neurite outgrowth in PC12 cells. PMID- 16697065 TI - Gamma histograms for radiotherapy plan evaluation. AB - BACKGROUND AND PURPOSE: The technique known as the 'gamma evaluation method' incorporates pass-fail criteria for both distance-to-agreement and dose difference analysis of 3D dose distributions and provides a numerical index (gamma) as a measure of the agreement between two datasets. As the gamma evaluation index is being adopted in more centres as part of treatment plan verification procedures for 2D and 3D dose maps, the development of methods capable of encapsulating the information provided by this technique is recommended. PATIENTS AND METHODS: In this work the concept of gamma index was extended to create gamma histograms (GH) in order to provide a measure of the agreement between two datasets in two or three dimensions. Gamma area histogram (GAH) and gamma volume histogram (GVH) graphs were produced using one or more 2D gamma maps generated for each slice of the irradiated volume. GHs were calculated for IMRT plans, evaluating the 3D dose distribution from a commercial treatment planning system (TPS) compared to a Monte Carlo (MC) calculation used as reference dataset. RESULTS: The extent of local anatomical inhomogenities in the plans under consideration was strongly correlated with the level of difference between reference and evaluated calculations. GHs provided an immediate visual representation of the proportion of the treated volume that fulfilled the gamma criterion and offered a concise method for comparative numerical evaluation of dose distributions. CONCLUSIONS: We have introduced the concept of GHs and investigated its applications to the evaluation and verification of IMRT plans. The gamma histogram concept set out in this paper can provide a valuable technique for quantitative comparison of dose distributions and could be applied as a tool for the quality assurance of treatment planning systems. PMID- 16697064 TI - Rapid screening of the fermentation profiles of wine yeasts by Fourier transform infrared spectroscopy. AB - A rapid screening method for the evaluation of the major fermentation products of Saccharomyces wine yeasts was developed using Fourier transform infrared spectroscopy and principal component factor analysis. Calibration equations for the quantification of volatile acidity, glycerol, ethanol, reducing sugar and glucose concentrations in fermented Chenin blanc and synthetic musts were derived from the Fourier transform infrared spectra of small-scale fermentations. The accuracy of quantification of volatile acidity in both Chenin blanc and synthetic must was excellent, and the standard error of prediction was 0.07 g l(-1) and 0.08 g l(-1), respectively. The respective standard error of prediction in Chenin blanc and synthetic musts for ethanol was 0.32% v/v and 0.31% v/v, for glycerol was 0.38 g l(-1) and 0.32 g l(-1), for reducing sugar in Chenin blanc must was 0.56 g l(-1) and for glucose in synthetic must was 0.39 g l(-1). These values were in agreement with the accuracy obtained by the respective reference methods used for the quantification of the components. The screening method was applied to quantify the fermentation products of glycerol-overproducing hybrid yeasts and commercial wine yeasts. Principal component factor analysis of the fermentation data facilitated an overall comparison of the fermentation profiles (in terms of the components tested) of the strains. The potential of Fourier transform infrared spectroscopy as a tool to rapidly screen the fermentative properties of wine yeasts and to speed up the evaluation processes in the initial stages of yeast strain development programs is shown. PMID- 16697066 TI - Predictive model for growth of Clostridium perfringens in cooked cured pork. AB - Mathematical models have been developed and used for predicting growth of foodborne pathogens in various food matrices. However, these early models either used microbiological media or other model systems to develop the predictive models. Some of these models have been shown to be inaccurate for applications in meat and specific food matrices, especially under dynamic conditions, such as constantly changing temperatures that are encountered during food processing. The objective of this investigation was to develop a model for predicting growth of Clostridium perfringens from spore inocula in cured pork ham. Isothermal growth of C. perfringens at various temperatures from 10 to 48.9 degrees C were evaluated using a methodology that employed a numerical technique to solve a set of differential equations. The estimated theoretical minimum and maximum growth temperatures of C. perfringens in cooked cured pork were 13.5 and 50.6 degrees C, respectively. The kinetic and growth parameters obtained from this study can be used in evaluating growth of C. perfringens from spore populations during dynamically changing temperature conditions such as those encountered in meat processing. Further, this model can be successfully used to design microbiologically "safe" cooling regimes for cured pork hams and similar products. PMID- 16697067 TI - Advances in the production of sponge biomass Aplysina aerophoba--a model sponge for ex situ sponge biomass production. AB - Sponges are a promising source of organic compounds of potential interest regarding industrial and medical applications. For detailed studies on such compounds, large amounts of sponge biomass are required. Obtaining that is at present extremely difficult because most sponges are relatively rare in nature and their mass cultivation in the laboratory has not yet been accomplished. In this study the possibility of culturing Aplysina aerophoba fragments in laboratory was examined. While a substantial biomass increase was not yet observed, we achieved fragmented sponge tissue to develop into a functional sponge as a first success. PMID- 16697068 TI - Alendronate-loaded nanoparticles deplete monocytes and attenuate restenosis. AB - Systemic transient depletion of monocytes and macrophages by liposome encapsulated bisphosphonates (BPs), reduces neointimal formation in experimental restenosis. The aim of this study was to examine the antirestenotic effect of a polymeric nanoparticulate formulation containing the BP alendronate (ALN). The BP was successfully formulated in polylactide-co-glycolide (PLGA) nanoparticles (NP). ALN NP with negative charge, size of 223+/-64 nm, and high entrapment efficiency (55.1%) have been formulated. ALN NP exhibited a significant cytotoxic effect, in a dose-response relationship, on macrophage-like RAW264 cells in cell culture. Subcutaneously (SC) administrated ALN NP (1.5 mg/kg on days -1 and +6) resulted in a significant attenuation of neointima to media ratio (N/M) by 52.7% and stenosis by 39.7% 28 days after balloon injury in the hypercholesterolemic rabbit model. Moreover, a good correlation was found between macrophage abundance in the injured arteries and the extent of stenosis. ALN NP treatment resulted in the reduction of both interleukin-1beta and matrix metalloproteinases (2 and 9). It is concluded that a particulated dosage form of polymeric NP loaded with ALN reduce neointimal formation in vivo by systemic transient depletion of monocytes. PMID- 16697069 TI - The "inverted cup" -- a novel in vitro release technique for drugs in lipid formulations. AB - The aim of this study was to develop a membrane-free in vitro release method for drugs in lipid formulations. It was intended to be applicable to as wide a range as possible of preparations, independently of their polarity and viscosity. The principle of the novel technique is to keep the sample suspended in the release medium in an inverted glass cup, allowing a possible phase transition or swelling. Thirteen formulations containing bupivacaine, lidocaine and/or prilocaine in lipid vehicles with different physical properties were prepared and examined. When possible, in vitro release profiles obtained by the new method were compared to profiles obtained by earlier techniques. For three formulations of either bupivacaine or lidocaine in polar lipid formulations, in vitro release profiles were evaluated in relation to in vivo data, from nerve block and pharmacokinetic studies in rats. Preparations that could be investigated both by the "inverted cup" and by the earlier published "single drop" technique generally showed good agreement between the two release profiles. In the case of the polar lipid formulations, arterial blood concentration curves in rats could reasonably be predicted from the in vitro release profiles. In conclusion, the "inverted cup" technique should potentially be applicable to a wide range of lipid formulations of drugs, both for physico-chemical characterisation and for obtaining in vitro -- in vivo correlations. PMID- 16697070 TI - Coping with the quantitative genomics 'elephant': the correlation between the gene dispensability and evolution rate. AB - Early studies suggested that proteins with a greater contribution to the fitness of an organism evolve more slowly than less 'important' proteins. Recent articles by two research groups highlight the long-standing controversy about the genome wide relationships between the measures of evolution rate, protein abundance and the fitness effect of gene disruption. These studies highlight the need for truly multidimensional approaches to the issues of quantitative genomics. PMID- 16697071 TI - Endophenotypes for psychiatric disorders: ready for primetime? AB - It is increasingly accepted that the imprecision of categorical psychiatric diagnoses can be a limiting factor in understanding the genetic basis of human behavioral abnormalities. Genetic investigation of endophenotypes--more precisely defined quantitative traits hypothesized to underlie disease syndromes--offers great promise as an alternative or complement to studies of categorical disease phenotypes. However, there is not yet standardization of the methods by which candidate endophenotypes should be chosen and applied. Fruitful endophenotype studies depend on the selection of heritable, quantitative traits that can be objectively and reliably measured. In this article, we propose guidelines for such investigations for psychiatric disorders, using endophenotypes previously proposed for bipolar disorder as particular examples. Gene expression studies and non-human primate models are recent developments in which an endophenotype approach might prove particularly valuable. PMID- 16697072 TI - Epigenetics and twins: three variations on the theme. AB - Twin studies have had a key role in the evaluation of heritability, a population based estimate of the genetic contribution to phenotypic variation. These studies have led to the revelation that most normal and disease phenotypes are to some extent heritable. Recently, interest has shifted from phenomenological heritability to the identification of trait-specific genes. The era of twin studies, however, is not over: recent epigenetic and global gene expression studies suggest that the most interesting findings in twin-based research are still to come. The increasing realization of the influence of epigenetics in phenotypic outcomes means that the molecular mechanisms behind phenotypic differences in genetically identical organisms can be explored. Analyses of epigenetic twin differences and similarities might yet challenge the fundamental principles of complex biology, primarily the dogma that complex phenotypes result from DNA sequence variants interacting with the environment. PMID- 16697073 TI - Large cell neuroendocrine carcinoma of the lung: a retrospective analysis of 144 surgical cases. AB - OBJECTIVE: Large cell neuroendocrine carcinoma of the lung are considered aggressive. However, reported prognoses are heterogeneous and the optimum treatment remains undefined. We retrospectively evaluated outcomes in a series of patients with a pathological diagnosis of large cell neuroendocrine lung carcinoma, who underwent lung resection. We also assessed the utility of chemotherapy in a small subgroup. PATIENTS AND METHODS: The clinical records of 144 consecutive patients were reviewed in a multicenter study. Survival times, assessed from the day of surgery until death or most recent follow-up, were estimated by the Kaplan-Meier method, and compared by the log rank test. RESULTS: There were 117 men and 27 women of median age 63 years. Twelve wedge resections, 3 segmentectomies, 95 lobectomies, 7 bilobectomies and 24 pneumonectomies were performed. Induction chemotherapy was given in 21 and postoperative chemotherapy in 24. Pathologically, 73 (50%) were stage I, 29 (20%) stage II, 40 (28%) stage III and 2 stage IV. Postoperative mortality was 2.8% and morbidity 26%. Overall 5 year survival was 42.5%: 52% for stage I, 59% for stage II and 20% for stage III (p=0.001 log-rank test on Kaplan-Meier curves). A trend to better outcome was associated with preoperative or postoperative chemotherapy in stage I disease (p=0.077) compared to no chemotherapy. The response rate to induction chemotherapy was 80% in the 15 patients with data available. CONCLUSION: large cell neuroendocrine carcinoma of the lung are confirmed as aggressive but are also chemosensitive. Our experience suggests that chemotherapy may improve prognosis in stage I disease. PMID- 16697074 TI - Prognostic value of expression of vascular endothelial growth factor and its flt 1 and KDR receptors in stage I non-small-cell lung cancer. AB - BACKGROUND: Angiogenesis plays an important role in tumorigenesis and has attracted interest as a potential target in cancer treatment. PATIENTS AND METHODS: We examined the prognostic value of the expression of vascular endothelial growth factor (VEGF) and of the VEGF receptors (VEGFRs) fms-like tyrosine kinase receptor-1 (flt-1) and kinase insert domain-containing receptor (KDR) in non-small-cell lung cancer (NSCLC). Sixty patients with surgical stage I NSCLC who had not undergone induction therapy or adjuvant therapy were selected from among 170 patients with NSCLC who had undergone surgery from January to December 1995. Specimens obtained at surgical resection were subjected to immunohistological staining, and the relationship between postoperative outcome and the expression of VEGF and its receptors was investigated. All patients included in the analysis had been followed up for 5 years or longer or until death. RESULTS: Patients with tumors expressing VEGF or KDR tended to have poorer outcomes, and VEGF expression and KDR expression were positively correlated. In contrast, flt-1 expression was not correlated with VEGF expression or outcome. Outcomes were poor in patients with tumors positive for both VEGF and VEGFRs. Multivariate analysis identified expression of both flt-1 and KDR and VEGF and KDR as possible independent prognostic factors. CONCLUSIONS: Our results suggest that expression of VEGF and VEGFR are associated with a poor prognosis via autocrine and paracrine growth stimulation of cancer cells. Moreover, tumors expressing both flt-1 and KDR may have greater malignant potential and are associated with a poor prognosis. PMID- 16697075 TI - Computer modelling in combination with in vitro studies reveals similar binding affinities of Drosophila Crumbs for the PDZ domains of Stardust and DmPar-6. AB - Formation of multiprotein complexes is a common theme to pattern a cell, thereby generating spatially and functionally distinct entities at specialised regions. Central components of these complexes are scaffold proteins, which contain several protein-protein interaction domains and provide a platform to recruit a variety of additional components. There is increasing evidence that protein complexes are dynamic structures and that their components can undergo various interactions depending on the cellular context. However, little is known so far about the factors regulating this behaviour. One evolutionarily conserved protein complex, which can be found both in Drosophila and mammalian epithelial cells, is composed of the transmembrane protein Crumbs/Crb3 and the scaffolding proteins Stardust/Pals1 and DPATJ/PATJ, respectively, and localises apically to the zonula adherens. Here we show by in vitro analysis that, similar as in vertebrates, the single PDZ domain of Drosophila DmPar-6 can bind to the four C-terminal amino acids (ERLI) of the transmembrane protein Crumbs. To further evaluate the binding capability of Crumbs to DmPar-6 and the MAGUK protein Stardust, analysis of the PDZ structural database and modelling of the interactions between the C-terminus of Crumbs and the PDZ domains of these two proteins were performed. The results suggest that both PDZ domains bind Crumbs with similar affinities. These data are supported by quantitative yeast two-hybrid interactions. In vivo analysis performed in cell cultures and in the Drosophila embryo show that the cytoplasmic domain of Crumbs can recruit DmPar-6 and DaPKC to the plasma membrane. The data presented here are discussed with respect to possible dynamic interactions between these proteins. PMID- 16697076 TI - Dual effects of staurosporine on A431 and NRK cells: microfilament disassembly and uncoordinated lamellipodial activity followed by cell death. AB - The general protein kinase inhibitor staurosporine (STS) has dual effects on human epidermoid cancer cells (A431) and normal rat kidney fibroblasts (NRK). It almost immediately stimulated increased lamellipodial activity of both cell lines and after 2 h induced typical signs of apoptosis, including cytoplasmic condensation, nuclear fragmentation, caspase-3 activation and DNA degradation. In the early phase we observed disruption of actin-containing stress fibres and accumulation of monomeric actin in the perinuclear region and cell nucleus. Increased lamellipodial-like extensions were observed particularly in A431 cells as demonstrated by co-localisation of actin and Arp2/3 complex, whereas NRK cells shrunk and exhibited numerous thin long extensions. These extensions exhibited uncoordinated centrifugal motile activity that appeared to tear the cells apart. Both cofilin and ADF were translocated from perinuclear regions to the cell cortex and, as expected in the presence of a kinase inhibitor, all the cofilin was dephosphorylated. Myosin II was absent from the extensions, and a reduction of phosphorylated myosin light chains was observed within the cytoplasm indicating myosin inactivation. Microtubules and intermediate filaments retained their characteristic filamentous organisation after STS exposure even when the cells became rounded and disorganised. Simultaneous treatment of NRK cells with STS and the caspase inhibitor zVAD did not inhibit the morphological and cytoskeletal changes. However, the cells underwent cell death as verified by positive annexin-V-staining. Thus it seems likely that cell death induced by STS may not only be a consequence of the activation of caspase, instead the disruption of the many motile processes involving the actin cytoskeleton may by itself suffice to induce caspase-independent cell death. PMID- 16697077 TI - Identification of two phosphatidylinositol/phosphatidylcholine transfer protein genes that are predominately transcribed in the flowers of Arabidopsis thaliana. AB - The Sec14 protein (Sec14p) and its homologs are involved in the transfer of phosphatidylinositol/phosphatidylcholine phospholipids in eukaryotic cells. In the completely sequenced genome of Arabidopsis thaliana, multiple genes encoding putative Sec14p homologs have been predicted based on bioinformatic analysis. Here we report the identification of two yeast Sec14-like genes (designated as AtSFH3 and AtSFH12, respectively) that are predominately transcribed in Arabidopsis flowers. The deduced amino acid sequences of AtSfh3p and AtSfh12p exhibited high similarity to that of Sec14p. Ectopic expression of AtSfh3p or AtSfh12p corrected the high temperature sensitive phenotype caused by Sec14p functional deficiency in Saccharomyces cerevisiae, indicating that the two plant homologs are functional in the intracellular environment. AtSFH3 transcripts were detected in flowers, stems and immature siliques but not roots and leaves, with a relatively higher transcript level in the flowers. In contrast, AtSFH12 transcripts were only detectable in the flowers. Based on histochemical staining of beta-glucuronidase (GUS) activities in the transgenic Arabidopsis plants harboring promoter::GUS constructs, AtSFH3 transcription was first detected in the stigma papillae of the flowers at stage 11, and then in the pollen grains before and after fertilization. On the other hand, AtSFH12 transcription was only found in the mature and germinating pollen grains. The information from this study may provide useful clue for further analysis of the function of plant Sec14p homologs in the development of the male gametic cells and/or the fertilization process in higher plants. PMID- 16697078 TI - CART peptide diurnal variations in blood and brain. AB - The central role of CART peptide in feeding, drug abuse and stress has been widely researched however, CART's role in the peripheral system are less explored. CART peptide is present in a variety of peripheral tissues including sympathetic ganglion neurons, adrenal glands, gut, pancreas and blood. Studies that examined circulating CART demonstrated that the active fragment with a molecular weight of CART55-102 is present in the blood of rats and rhesus macaques. Interestingly, CART expression in these species exhibits a distinctive diurnal rhythm which correlates with the respective daily rhythms of corticosterone and feeding. In the rat, adrenalectomy significantly reduces blood CART levels and abolishes its daily rhythm while corticosterone replacement reinstates CART expression to control levels. In addition, direct administration of corticosterone significantly increases CART blood levels while administration of corticosterone synthesis blocker metyrapone, inhibits CART blood levels. These data suggest that the adrenal gland could be a source of blood CART and that glucocorticoids may play a role in the generation of CART's diurnal rhythm. Moreover, fuel availability may be important in the control of CART levels and its daily rhythm, since 24 h food restriction alters CART levels and abolishes its rhythm. In addition to blood, both CART peptide and mRNA exhibit food dependent diurnal rhythm in discrete rat brain areas including the nucleus accumbens, amygdala and hypothalamus. Altogether, these findings suggest that CART is influenced by hypothalamic-pituitary-adrenal interactions and that it may play a role in multiple physiological processes possibly involving feeding, stress, reward and motivation. PMID- 16697079 TI - Long-term effects of nicotine on the forced swimming test in mice: an experimental model for the study of depression caused by smoke. AB - Large evidence showing an association between depression and tobacco smoking is known. Nicotine is the active chemical responsible for smoking addiction, and its withdrawal may induce in smokers greater sensitivity to stress. Our aim has been to investigate the links between tobacco addiction and depression by studying the long-term effects of repeated administration of nicotine followed by dependence, to forced swimming test, serotonin content and 5-HT(1A) expression in diencephalon. Dependence has been induced by daily subcutaneous injection in mice of nicotine (2mg/kg four injections daily) for 15 days and assessed after nicotine withdrawal with an abstinence scale; control animals received daily subcutaneous injection of saline for the same period. Experiments on forced swimming test have been carried out at t=0 (last day of nicotine or saline treatment), and 15, 30, 45 and 60 days after saline or nicotine withdrawal. Both control mice and nicotine mice have been pre-treated with oral 5-hydroxy tryptophan (12.5-50mg/kg), precursor of serotonin, before forced swimming test. Nicotine mice have shown on forced swimming test a significant increase of immobility time compared to control mice. This increase was not evident in nicotine mice treated with 5-hydroxy-tryptophan and treatment with the selective serotonin receptorial antagonist WAY 100635 (WAY) abolished 5-hydroxy-tryptophan effects. Evaluation of diencephalic serotonin, performed at t=0 showed an increase of diencephalic serotonin content, while serotonin measured 15, 30, 45 and 60 days after nicotine withdrawal, was significantly reduced in nicotine mice compared to control mice. Western blot analysis showed a great reduction of 5 HT(1A) receptor expression in nicotine mice measured at t=0 (last day of treatment) and at 15 and 30 days after nicotine withdrawal compared to control mice. Our results show that (i) behavioural alterations estimated with forced swimming test and (ii) changes in diencephalic serotonin content and 5-HT(1A) receptor expression, are present since nicotine is withdrawn even after a long time, suggesting a role of serotonin in mood disorders eventually occurring following smoking cessation. PMID- 16697080 TI - Novel potent and selective alphavbeta3/alphavbeta5 integrin dual antagonists with reduced binding affinity for human serum albumin. AB - The binding of lead compounds and drugs to human serum albumin (HSA) is a ubiquitous problem in drug discovery since it modulates the availability of the leads and drugs to their intended target, which is linked to biological efficacy. In our continuing efforts to identify small molecule alpha(V)beta(3) and alpha(V)beta(5) dual antagonists, we recently reported indoles 2-4 as potent and selective alpha(V)beta(3)/alpha(V)beta(5) antagonists with good oral bioavailability profile. In spite of subnanomolar binding affinity of these compounds to human alpha(V)beta(3) and alpha(V)beta(5) integrins, high HSA binding (96.5-97.3%) emerged as a limiting feature for these leads. Structure activity HSA binding data of organic acids reported in the literature have demonstrated that the incorporation of polar groups into a given molecule can dramatically decrease the affinity toward HSA. We sought to apply this strategy by examining the effects of such modifications in both the central core constrain and the substituent beta to the carboxylate. Most of these derivatives were prepared in good yields through a cesium fluoride-catalyzed coupling reaction. This reaction was successful with a variety of nitrogen-containing scaffolds (20, 33, and 43) and selected acetylenic derivatives (16, 19, and 34). Among the compounds synthesized, the 3-[5-[2-(5,6,7,8-tetrahydro [1,8]naphthyridin-2 yl)ethoxy]indol-1-yl]-3-[5-(N,N-dimethylaminomethyl)-3-pyridyl]propionic acid (25) was found to be the most promising derivative within this novel series with a subnanomolar affinity for both alpha(v)beta(3) and alpha(v)beta(5) (IC(50) = 0.29 and 0.16 nM, respectively), similar to our initial lead receptor antagonists 2-4, and exhibiting a low HSA protein binding (40% bound, K(d) = 1.1+/-0.4 x 10(3) microM) and an improved in vitro stability profile toward human and mouse microsomes (99.9% and 98.7% remaining after 10 min). Moreover, the selectivity of 25 toward alpha(5)beta(1) and IIbIIIa integrins was perfectly maintained when compared to the parent leads 2-4. Thus, compound 25 was selected as a new lead with improved drug-like properties for further evaluations in the field of oncology and osteoporosis. PMID- 16697081 TI - [The unexpected Chikungunya outbreak]. PMID- 16697082 TI - [Endobronchial stenosis in Wegener's granulomatosis]. AB - INTRODUCTION: Endobronchial stenoses are rare in the course of Wegener's granulomatosis, and they occur even more rarely than subglottic stenoses. EXEGESIS: We report seven cases of endobronchial stenoses in the setting of Wegener granulomatosis. Neither the pulmonary symptoms nor the systemic manifestations of vasculitis were specific. However 6/7 patients presented a wheezing or an hemoptysis. Bronchial endoscopy has permitted the diagnosis in all cases. Local evolution was cicatricial and symptomatic stenosis in 3 cases (42,8%). CONCLUSION: Thus these lesions must be research in any case of pulmonary abnormality in the course of Wegener's granulomatosis, because they may lead to a pejorative prognosis. Moreover general and local treatment must be given early (at the inflammatory stage). After this stage, the local treatments are difficult and not efficient. PMID- 16697083 TI - [Iatrogenic angioedema due to estrogen, angiotensin conversion enzyme inhibitors, angiotensin receptor antagonists and dialysis membranes]. PMID- 16697084 TI - ESPEN Guidelines on Enteral Nutrition: Cardiology and pulmonology. AB - These guidelines are intended to give evidence-based recommendations for the use of enteral nutrition (EN) in patients with chronic heart failure (CHF) and chronic obstructive pulmonary disease (COPD). They were developed by an interdisciplinary expert group in accordance with officially accepted standards and are based on all relevant publications since 1985. They have been discussed and accepted in a consensus conference. EN by means of oral nutritional supplements (ONS) or tube feeding (TF) enables nutritional intake to be maintained or increased when normal oral intake is inadequate. No data are yet available concerning the effects of EN on cachexia in CHF patients. However, EN is recommended to stop or reverse weight loss on the basis of physiological plausibility. In COPD patients, EN in combination with exercise and anabolic pharmacotherapy has the potential to improve nutritional status and function. Frequent small amounts of ONS are preferred in order to avoid postprandial dyspnoea and satiety as well as to improve compliance. PMID- 16697085 TI - Evidence supports nutritional support. PMID- 16697086 TI - Introductory to the ESPEN Guidelines on Enteral Nutrition: Terminology, definitions and general topics. AB - The ESPEN guidelines on enteral nutrition are the first evidence-based European recommendations for enteral nutrition. They were established by European experts for a variety of disease groups. During guideline development it became evident that terms and definitions in clinical nutrition have been used inconsistently depending on medical disciplines as well as regional and personal preferences. Therefore, to increase explanatory accuracy it was necessary to unify them. In this chapter terms and definitions used throughout all guidelines are explained. Additionally answers to more general questions, which might be important in most indications are dealt with, i.e. use of fibre containing and diabetes formulae. PMID- 16697087 TI - ESPEN Guidelines on Enteral Nutrition: Intensive care. AB - Enteral nutrition (EN) via tube feeding is, today, the preferred way of feeding the critically ill patient and an important means of counteracting for the catabolic state induced by severe diseases. These guidelines are intended to give evidence-based recommendations for the use of EN in patients who have a complicated course during their ICU stay, focusing particularly on those who develop a severe inflammatory response, i.e. patients who have failure of at least one organ during their ICU stay. These guidelines were developed by an interdisciplinary expert group in accordance with officially accepted standards and are based on all relevant publications since 1985. They were discussed and accepted in a consensus conference. EN should be given to all ICU patients who are not expected to be taking a full oral diet within three days. It should have begun during the first 24h using a standard high-protein formula. During the acute and initial phases of critical illness an exogenous energy supply in excess of 20-25 kcal/kg BW/day should be avoided, whereas, during recovery, the aim should be to provide values of 25-30 total kcal/kg BW/day. Supplementary parenteral nutrition remains a reserve tool and should be given only to those patients who do not reach their target nutrient intake on EN alone. There is no general indication for immune-modulating formulae in patients with severe illness or sepsis and an APACHE II Score >15. Glutamine should be supplemented in patients suffering from burns or trauma. PMID- 16697088 TI - Intramuscular immunization with a DNA vaccine encoding a 26-amino acid CETP epitope displayed by HBc protein and containing CpG DNA inhibits atherosclerosis in a rabbit model of atherosclerosis. AB - Rabbits were intramuscularly immunized with the plasmid pCR-X8-HBc-CETP encoding a B-cell epitope of cholesteryl ester transfer protein (CETP) C-terminal fragment (CETPC) displayed by Hepatitis B virus core (HBc) particle. This plasmid also contained immunostimulatory sequences (ISS) which included eight CpG motifs 5' GACGTT-3', functioning as immunomodulators. After anti-CETP antibodies were successfully produced, rabbits were fed with a high-cholesterol diet for 15 weeks, and then the antiatherogenic effects of this DNA immunization were evaluated. The results showed that the fraction of plasma cholesterol in HDL significantly increased and the fraction of plasma cholesterol in LDL decreased in the pCR-X8-HBc-CETP immunized rabbits compared with those in the saline control group and one group treated with the plasmid pCR-X8-HBc containing ISS but lacking CETP epitope. More importantly, DNA immunization with pCR-X8-HBc-CETP markedly reduced the average percentage of aortic lesions in the entire aorta area by 80.60% compared with the saline control (3.78% versus 19.48%) and the average thickness of hyperplastic coronary artery in this group was also significantly less than in the saline control group (146+/-11 microm versus 248+/ 18 microm). Our data also showed that CpG DNA alone could be antiatherogenic in this model because the average percentage of aortic lesions in pCR-X8-HBc immunized rabbits was 16.53% lower than that of the saline control group and the average thickness of hyperplastic coronary artery was also substantially lower than saline control group (155+/-13 microm versus 248+/-18 microm). Thus, plasmid pCR-X8-HBc-CETP could significantly inhibit the progression of atherosclerosis and be potentially developed as a suitable DNA vaccine against atherosclerosis. PMID- 16697089 TI - A vaccine against the salmonid pathogen Piscirickettsia salmonis based on recombinant proteins. AB - We report here the protective effect against piscirickettsiosis elicited in fish by a mixture of recombinant proteins. A comparative genomics strategy was used on a genomic library of Piscirickettsia salmonis in order to select optimal candidates for a recombinant subunit vaccine to protect fish from rickettsial septicaemia (SRS). Based on this information, 15 P. salmonis ORFs encoding heat shock proteins, virulence factors, membrane bound and other surface exposed antigens, were isolated and expressed. Seven of the most promising antigens were formulated in three mixtures (V1-V3) containing two or three recombinant proteins each and injected into salmon to test their protective efficacy. Two of the three formulations (V1, V2) elicited a strong protective response in a challenge against the pathogen, which was coincident with the humoral response against the corresponding recombinant proteins present in each formulation. V1, formulated with recombinant chaperonines Hsp60, Hsp70 and flagellar protein FlgG of P. salmonis achieved the highest level of protection with a relative percent survival (RPS) of 95%. PMID- 16697090 TI - Evaluation of novel Brucella melitensis unmarked deletion mutants for safety and efficacy in the goat model of brucellosis. AB - Pregnant goats were employed to assess unmarked deletion mutant vaccine candidates BMDeltaasp24, BMDeltacydBA, and BMDeltavirB2, as the target host species naturally infected with Brucella melitensis. Goats were assessed for the degree of pathology associated with the vaccine strains as well as the protective immunity afforded by each strain against abortion and infection after challenge with wild-type Brucella melitensis 16M. Both BMDeltaasp24 and BMDeltavirB2 were considered safe vaccine candidates in the pregnant goat model because they did not cause abortion or colonize fetal tissues. BMDeltaasp24 was isolated from the maternal tissues only, indicating a slower rate of clearance of the vaccine strain than for BMDeltavirB2, which was not isolated from any maternal or fetal tissues. Both strains were protective against abortion and against infection in the majority of pregnant goats, although BMDeltaasp24 was more efficacious than BMDeltavirB2 against challenge infection. PMID- 16697091 TI - Immunization against Japanese encephalitis in China: a policy analysis. AB - This study examined the impact of Japanese encephalitis (JE) immunization policies on disease trends in China. Through a document review and key source interviews in eight provinces, the study found, JE immunizations were provided via a combination of fee-for-service and government funding. Unequal government funding and unbalanced economic development led to variation in JE incidence levels between provinces. Government support of low-fee JE immunization, has led to nationwide decline of JE incidence by >90%; however, greater reliance on user fees and market mechanisms in rural areas has limited the provision of JE and other childhood immunizations to poor rural children. PMID- 16697092 TI - The seeking of baby-feeding information by Canadian women pregnant with twins. AB - OBJECTIVE: To analyse baby-feeding information needs and seeking described by Canadian women pregnant with twins. DESIGN: Qualitative; data were collected by in-depth semi-structured interviews. Transcripts provided the data for discourse analysis of the use of two interpretative repertoires. SETTING: Ontario, Canada. PARTICIPANTS: 19 women pregnant with twins. MEASUREMENTS AND FINDINGS: The first interpretative repertoire represented caring for twins as fundamentally distinct from caring for singly born babies, and therefore emphasised the commonality of mothers of twins regardless of their background or situation. The second highlighted the uniqueness and individuality of each person. These repertoires intersect with discourses of baby feeding and good mothering, resulting in a complex discursive interplay of similarity and difference, commonality and individuality, information seeking, baby feeding and good mothering. Participants used the two interpretative repertoires to (1) frame information needs; (2) construct complex accounts of the biomedical, experiential and personalised authority of information sources; (3) describe the helpfulness of information sources independent of their authority; and (4) describe the barriers encountered in the finding appropriate sources of baby-feeding information in pregnancy. IMPLICATIONS: Understanding and respecting the discursive constructions of pregnancy and mothering can help practitioners understand the complex discursive interplay underlying participants' baby-feeding decisions, and may facilitate more sensitive support for women's individual needs and understandings. PMID- 16697093 TI - Influence of bacteria on Pb and Zn speciation, mobility and bioavailability in soil: A laboratory study. AB - A soil column experiment was carried out to investigate the effects of inoculation of bacteria on metal bioavailability, mobility and potential leachability through single chemical extraction, consequential extraction and in situ soil solution extraction technologies. Results showed that bacteria inoculated, including Azotobacter chroococcum, Bacillus megaterium and Bacillus mucilaginosus, may pose both positive and negative impacts on bioavailability and mobility of heavy metals in soil, depending on the chemical nature of the metals. The activities of bacteria led to an increase of water dissolved organic carbon (DOC) concentration and a decrease of pH value, which enhanced metal mobility and bioavailability (e.g. an increase of water-soluble and HOAc-soluble Zn). On the other hand, bacteria could immobilize metals (e.g. a great reduction of water soluble Pb) due to the adsorption by bacterial cell walls and possible sedimentation reactions with phosphate or other anions produced through bacterial metabolism. PMID- 16697094 TI - Affective empathy deficits in aggressive children and adolescents: a critical review. AB - Aggressive children and adolescents are often hypothesized to have lower levels of empathy than comparable nonaggressive peers. A review of 17 studies examining the relationship between affective empathy and aggressive or delinquent behavior was conducted to determine the validity of this hypothesis. The studies offered conflicting findings, even when the measures of empathy were identical across studies. Based on this review, there was not a consistent relationship between empathy and aggression in children, but a negative relationship was typically found in adolescents. Self-report measures of empathy used with adolescent participants showed the most robust relationship with aggression. Clinical implications, limitations of present research, and recommendations for future research are discussed. PMID- 16697095 TI - Communicative and strategic action in interpreted consultations in primary health care: a Habermasian perspective. AB - We report a qualitative study of accounts of interpreted consultations in UK primary care. The study sought to explore how three Habermasian tensions between (a) system and lifeworld, (b) communicative and strategic action, and (c) interpersonal and macropolitical spheres played out in the triadic consultation between clinician, interpreter and patient. In a total of 69 individual interviews and two focus groups, we collected narratives from service users (through interpreters or bilingual researchers), interpreters and doctors and other staff in general practice. We recorded, transcribed and analysed these, taking the story as the main unit of analysis. Our data suggest that the preconditions for communicative action are rarely met in the interpreted consultation. The interpreter's presence makes a dyadic interaction into a triad, adding considerable complexity to the social situation and generating operational and technical challenges. Lack of trust, intense pressure of time, mismatch of agendas (biomedical versus lifeworld), firm expectations of a specific outcome (e.g. referral, prescription) and profound power imbalances all promote strategic action (i.e. speech that seeks consciously or unconsciously to manipulate an outcome) rather than communicative action (i.e. sincere efforts to achieve understanding, and reach consensus) by all parties. In consultations interpreted by family members (an option traditionally seen as 'second best' by policy makers), the social situation is very different. Family members are generally trusted, share the lifeworld agenda, and shift the power balance in the patient's favour. The interpreter occupies multiple social roles, including translator, interpersonal mediator, system mediator, educator, advocate, and link worker. The essence of professionalism in interpreting is shifting judiciously between these potentially conflicting roles. We discuss the implications of our findings for communication with limited English speakers in healthcare consultations and for realizing contemporary policy goals such as concordance, shared decision-making, empowerment, and choice. PMID- 16697096 TI - A typology of preferences for participation in healthcare decision making. AB - Classifying patients as "active" or "passive" with regard to healthcare decision making is misleading, since patients have different desires for different components of the decision-making process. Distinguishing patients' desired roles is an essential step towards promoting care that respects and responds to individual patients' preferences. We included items on the 2004 Wisconsin Longitudinal Study mail survey measuring preferences for four components of the decision-making process: physician knowledge of patient medical history, physician disclosure of treatment choices, discussion of treatment choices, and selection of treatment choice. We characterized preference types for 5199 older adults using cluster analysis. Ninety-six percent of respondents are represented by four preference types, all of which prefer maximal information exchange with physicians. Fifty-seven percent of respondents wanted to retain personal control over important medical decisions ("autonomists"). Among the autonomists, 81% preferred to discuss treatment choices with their physician. Thirty-nine percent of respondents wanted their physician to make important medical decisions ("delegators"). Among the delegators, 41% preferred to discuss treatment choices. Female gender, higher educational attainment, better self-rated health, fewer prescription medications, and having a shorter duration at a usual place of care predicted a significantly higher probability of the most active involvement in discussing and selecting treatment choices. The overwhelming majority of older adults want to be given treatment options and have their physician know everything about their medical history; however, there are substantial differences in how they want to be involved in discussing and selecting treatments. PMID- 16697097 TI - Expression changes of neurofilament subunits in the central nervous system of hens treated with tri-ortho-cresyl phosphate (TOCP). AB - Tri-ortho-cresyl phosphate (TOCP) could induce degeneration of long, large diameter axons within the central and peripheral nervous system of susceptible species including human being and hens, which is referred to as organophosphorus ester induced delayed neuropathy (OPIDN). The mechanisms involved are not understood. Neuropathologic observations suggested that neurofilament subunits (NFs) could be a main target of TOCP in the peripheral nervous system. Our previous study also showed that NFs in protein levels significantly decreased in sciatic nerves of hens treated with TOCP. In this study, to determine whether the decrement of NFs proteins in sciatic nerves was due to reductions in NF gene expression or protein degradation, hens were treated with a single dose of 750 mg/kg body weight TOCP by gavage, and sacrificed on 21 day post-exposure. Cerebral cortexes and spinal cords were sampled. Transcriptional changes of NFs including high molecular weight neurofilament (NF-H), middle molecular weight neurofilament (NF-M), low molecular weight neurofilament (NF-L), and glyceraldehydes-3-phoaphate dehydrogenase (GAPDH) as inner inference in cerebral cortexes and spinal cords were analyzed by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Results showed that all of three NFs mRNA in cerebral cortexes down-regulated significantly. However, in spinal cords, there was only NF-M decreased, both of NF-H and NF-L kept unaffected. The protein levels of NFs in pellet and supernatant fractions of cerebral cortexes and spinal cords were also determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. We noticed that all NFs protein declined in pellet of cerebral cortexes, but NF-M reduction was not significant compared with that of control hens. NF-H and NF-M proteins in supernatant of cerebral cortexes exhibited significant increase, while NF-L level showed remarkable decline. In spinal cords, apart from NF-L in pellet were significantly increased, both of NF-H and NF-M in pellet and supernatant, as well as NF-L in supernatant fractions were manifested dramatic reduction compared with the pattern of control. The quantitative analyses revealed that the change magnitude in protein levels was much greater than that in mRNA levels in hens' central nervous system after TOCP administration. These findings suggest that the NFs disturbance in protein levels is closely associated with the decreases in sciatic nerves observed in our previous work after TOCP exposure, rather than that in mRNA levels, and the NFs alterations in protein levels may be one of the responsible factors for the OPIDN. PMID- 16697098 TI - Urinary biomarkers of di-isononyl phthalate in rats. AB - Commercial di-isononyl phthalate (DiNP) is a mixture of various branched-chain dialkyl phthalates mainly containing nine-carbon alkyl isomers. At high doses in rodents, DiNP is a carcinogen, and a developmental toxicant. After exposure, the diester isomers are de-esterified to form hydrolytic monoesters, monoisononyl phthalates (MiNP), which subsequently metabolize to form oxidative metabolites. These metabolites can be excreted in urine or feces. The urinary excretion of DiNP metabolites was monitored in adult female Sprague-Dawley rats after oral administration of a single dose (300 mg/kg) of commercial DiNP. The metabolites were extracted from urine, resolved with high performance liquid chromatography, analyzed by mass spectrometry, and tentatively identified based on their chromatographic separation and mass spectrometric fragmentation pattern. Because DiNP is an isomeric mixture, its metabolites were also isomeric mixtures that eluted from the HPLC column with close retention times. Mono(carboxy isooctyl)phthalate (MCiOP) was identified as the major metabolite of DiNP; in addition, mono(hydroxy-isononyl)phthalate (MHiNP) and mono(oxo-isononyl)phthalate (MOiNP) were present. Furthermore, metabolites of di-isooctyl phthalate (DiOP) and di-isodecyl phthalate (DiDP) were also detected. Excretion toxicokinetics of the DiNP metabolites in urine followed a biphasic pattern with initial rapid decay in concentration. Despite potential differences in the metabolism of DiNP among species, MCiOP, MHiNP and MOiNP were detected in humans with no known exposure to DiNP at levels significantly higher than MiNP suggesting that these oxidative metabolites may be better urinary biomarkers of human exposure to DiNP than is MiNP. PMID- 16697099 TI - Cantharidin-induced cytotoxicity and cyclooxygenase 2 expression in human bladder carcinoma cell line. AB - Mylabris is used in clinical therapy, but is always accompanied by cystitis. The toxic effects of mylabris on bladder are attributed to its active principle: cantharidin. In the present study, we explored how cantharidin induces cytotoxicity in the bladder. Human bladder carcinoma cell line T 24 cells were used as target cells, and human colon carcinoma HT 29 cells as native cells. Cantharidin exhibited acute cytotoxicity in the T 24 cells, and IC(50) was 21.8, 11.2 and 4.6 microM after treatment for 6, 24 and 48 h, respectively. The cytotoxicity of cantharidin was not significantly enhanced when T 24 cells were treated for a longer time. Moreover, PARP proteins and pro-caspase 3, Bcl-2 were significantly inhibited after cantharidin treatment in T 24 cells. Pretreatment with the caspase 3 inhibitor markedly inhibited cantharidin-induced cell death. Therefore, we suggested that cantharidin could induce apoptosis via active caspase 3 in T 24 cells. When T 24 cells were treated with cantharidin at a low dose, the cell cycle was arrested in the G(2)/M phase. Furthermore, p21(Cip1/Waf1) was enhanced, and cyclin A, B1 and cdk1 decreased. At a high dose (more 12.5 microM), cantharidin could stimulate T 24 cells to deplete a large number of ATP and induce secondary necrosis. In addition, cantharidin also stimulated COX 2 over-expression and PGE(2) production in T 24 cells, in a dose dependent manner. However, cantharidin also induced apoptosis and G(2)/M phase arrest in HT 29 cells, but did not induce COX 2 over-expression. Therefore, we suggest that cantharidin may induce cystitis through secondary necrosis and COX 2 over-expression. PMID- 16697100 TI - Conformational changes and activity alterations induced by nickel ion in horseradish peroxidase. AB - Conformational changes induced by the binding of nickel to horseradish peroxidase C (HRPC) were studied by electronic absorption spectroscopy, fluorescence spectroscopy and circular dichroism spectroscopy. Incubation of HRPC with various concentrations of Ni(2+) for 5 minutes resulted in changes in the enzyme absorption spectrum, including variations in the intensities of the Soret, beta and charge transfer (CT1) bands absorption, shift in the Soret, beta and CT1 bands maxima and absorption increase at 275 nm. Increases in the enzyme's intrinsic fluorescence as determined by fluorescence spectroscopy, as well as changes in the alpha-helical content, as determined by circular dichroism spectroscopy, were also found. Correlatively, alterations of the enzymatic activity by Ni(2+) were studied by following the H(2)O(2)-mediated oxidation of o dianisidine and 2,2'-azinobis(3-ethylbenzothiazolinesulfonic acid) (ABTS) by HRPC. With both reducing substrates, it was found that in the presence of sufficient amount of enzyme, 1-10 mM nickel would enhance the enzymatic activity, while higher Ni(2+) concentrations (20-50 mM) would inhibit it. The enzyme was completely inhibited after 5 minutes incubation in 50 mM Ni(2+). Prolonged incubation would induce complete inhibition at lower Ni(2+) concentrations. Spectrophotometry investigations also showed that inhibitory concentrations of Ni(2+) altered compounds I and II formation, compound II being the first affected. Based on spectrophotometry, fluorescence and circular dichroism spectroscopy, and data on compounds I and II formation, a scheme is suggested for HRPC conformational changes in different Ni(2+) concentrations. HRPC was found to have four potential attachment sites for Ni(2+) which were sequentially occupied in a dose- and time-dependent manner by the metallic ion. PMID- 16697101 TI - Characterization of the non-native trifluoroethanol-induced intermediate conformational state of the Shiga toxin B-subunit. AB - The effect of increasing concentrations of 2,2,2-trifluoroethanol (TFE) on the conformational stability of the Shiga toxin B-subunit (STxB), a bacterial homopentameric protein involved in cell-surface binding and intracellular transport, has been studied by far-, near-UV circular dichroism (CD), intrinsic fluorescence, analytical ultracentrifugation, and differential scanning calorimetry (DSC) under equilibrium conditions. Our data show that the native structure of STxB is highly perturbed by the presence of TFE. In fact, at concentrations of TFE above 20% (v/v), the native pentameric conformation of the protein is cooperatively transformed into a helix-rich monomeric and partially folded conformational state with no significant tertiary structure. Additionally, no cooperative transition was detected upon a further increase in the TFE concentration (above 40% (v/v)). The thermal stability of STxB was investigated at several different TFE concentrations using DSC and CD spectroscopy. Thermal transitions at TFE concentrations of up to 20% (v/v) were successfully fitted to the two-state folding/unfolding coupled to oligomerization model consistent with the transition between a pentameric folded conformation to a monomeric state of the protein, which the presence of TFE stabilizes as a partially folded conformation. PMID- 16697102 TI - P2X7 receptors in the nervous system. AB - P2X(7) receptors are ligand-gated ion channels, expressed as homo-oligomeric assemblies of individual subunits. They are widely distributed at immunocompetent cells of the central and peripheral nervous system and are believed to be primarily involved in host-defense reaction. However, a growing amount of evidence indicates that their signaling role in the brain is more widespread than previously anticipated. In this paper, we review the present knowledge on the structural and pharmacological features of the P2X(7) receptor, as well as its cell-type specific localization in the nervous system. Subsequently, the participation of P2X(7) receptors in distinct neuronal, astroglial and microglial functions are described. Finally, since they may play a prominent role in certain neurologic disorders, such as ischemia-reperfusion injury, Alzheimer's disease, spinal cord injury and sensory neuropathies, the pathological role and potential therapeutic exploitation of P2X(7) receptors are also discussed. PMID- 16697103 TI - Environmental risk perception, environmental concern and propensity to participate in organic farming programmes. AB - This paper aims to identify the factors underlying farmers' propensity to participate in organic farming programmes in a Romanian rural region that confronts non-point source pollution. For this, we employ structural equation modelling with latent variables using a specific data set collected through an agri-environmental farm survey in 2001. The model includes one 'behavioural intention' latent variable ('propensity to participate in organic farming programmes') and five 'attitude' and 'socio-economic' latent variables ('socio demographic characteristics', 'economic characteristics', 'agri-environmental information access', 'environmental risk perception' and 'general environmental concern'). The results indicate that, overall, the model has an adequate fit to the data. All loadings are statistically significant, supporting the theoretical basis for assignment of indicators for each latent variable. The significance tests for the structural model parameters show 'environmental risk perception' as the strongest determinant of farmers' propensity to participate in organic farming programmes. PMID- 16697104 TI - Stimulatory effects of lipopolysaccharide on endothelial cell activin and follistatin. AB - Activin A and its binding protein, follistatin, are released into the circulation following acute systemic inflammation. In this study, we determined the activin and follistatin response of ovine aortic endothelial cells to lipopolysaccharide (LPS). Exposure to LPS for 1h, mimicking a transient inflammatory event, elicited significant increases in activin betaA subunit mRNA or activin A release, with larger, more prolonged increases evident with continuous exposure. On the other hand, follistatin increases were only evident with prolonged exposure to LPS and following increases in activin A release. While cell-associated activin A increased with LPS exposure, levels were lower than those secreted, whereas the opposite was apparent for follistatin. In summary, our findings suggest that vascular endothelial cells, while capable of releasing activin A and follistatin following inflammatory stimulation, are unlikely to be responsible for the rapid release of activin A in vivo following inflammatory challenge. PMID- 16697105 TI - Evolution of arylalkylamine N-acetyltransferase: emergence and divergence. AB - The melatonin rhythm-generating enzyme, arylalkylamine N-acetyltransferase (AANAT) is known to have recognizable ancient homologs in bacteria and fungi, but not in other eukaryotes. Analysis of new cDNA and genomic sequences has identified several additional homologs in other groupings. First, an AANAT homolog has been found in the genome of the cephalochordate amphioxus, representing the oldest homolog in chordates. Second, two AANAT homologs have been identified in unicellular green algae. The homologs in amphioxus, unicellular green algae, fungi and bacteria are similarly primitive in that they lack sequences found in vertebrate AANATs that are involved in regulation and that facilitate binding and catalysis. In addition, all these sequences are intronless. These features are consistent with horizontal transfer of the AANAT ancestor from bacteria to green algae, fungi and chordates. Lastly, a third AANAT gene has been found in teleost fish, suggesting that AANAT genes serve multiple functions in addition to melatonin synthesis. PMID- 16697106 TI - Isocostunolide, a sesquiterpene lactone, induces mitochondrial membrane depolarization and caspase-dependent apoptosis in human melanoma cells. AB - Isocostunolide is a sesquiterpene lactone isolated from the roots of Inula helenium. Its chemical structure was determined by NMR and FAB-MS spectra. No biological activities of this compound have yet been reported. In this study, we found isocostunolide could effectively induce cytotoxicity in three cancer cell lines (A2058, HT-29, and HepG2), with an IC(50) of 3.2, 5.0, and 2.0 micro g/mL, respectively. DNA flow cytometric analysis indicated that isocostunolide actively induced apoptosis of cancer cells accompanied by a marked loss of G0/G1 phase cells. To address the mechanism of the apoptotic effect of isocostunolide, we analyzed the induction of apoptosis-related proteins in A2058. The levels of pro caspase-8, Bid, pro-caspase-3, and poly(ADP-ribose) polymerase (PARP) decreased. However, the level of Fas was increased markedly in a dose-dependent manner. Furthermore, this compound markedly induced a depolarization of mitochondrial membranes to facilitate cytochrome c release into cytosol. The findings suggest that isocostunolide may activate a mitochondria-mediated apoptosis pathway. To address this, we found that isocostunolide-induced loss of mitochondrial membrane potential occurred via modulation of the Bcl-2 family proteins. The production of intracellular reactive oxygen species (ROS) in A2058 was not elicited. In summary, for the first time, we have isolated and characterized isocostunolide from I. helenium. This compound induces apoptosis through a mitochondria dependent pathway in A2058 cells. PMID- 16697107 TI - Frequent epigenetic inactivation of p53 target genes in seminomatous and nonseminomatous germ cell tumors. AB - Hypermethylation of tumor-suppressor genes has been implicated in the pathogenesis of human cancers. This study was designed to examine the methylation profiles of a selected group of p53 target genes (APAF-1, CASP-8, DAPK-1, IGFBP 3) and to correlate the findings with the histopathological characterization of testicular germ cell tumors (TGCT). Promoter methylation status was analysed by highly sensitive real-time methylation-specific PCR in 46 primary TGCTs (26 seminomas and 20 nonseminomas) and 15 normal testicular tissue samples. APAF-1 methylation was detected in all of the seminomatous and nonseminomatous TGCTs as well as in 60% of normal testicular tissue. Methylation of DAPK-1 was frequent in seminomas (50%) and nonseminomas (20%), but not in normal testicular tissue (6%). The degree of DAPK-1 methylation correlated with the clinical stage of the disease (P=0.05) and was useful in differentiating seminomatous from nonseminomatous, and malignant from nonmalignant testicular tissue (P=0.04 and 0.02, respectively). The APAF-1 methylation index achieved a highly significant differentiation between seminomatous or nonseminomatous tissue and nonmalignant testicular tissue (P=0.0001). In testicular tumorigenesis, promoter methylation of specific p53 target genes occurs at early stage but to varying degrees. Methylation also occurs in normal testicular tissue, which is in contrast to findings in other urogenital malignancies. Further studies will be necessary to determine whether the methylation level may be used as marker for risk estimation, especially in clinical stage I disease. PMID- 16697108 TI - Amitriptyline suppresses neuroinflammation and up-regulates glutamate transporters in morphine-tolerant rats. AB - The present study was performed to evaluate the effects of the tricyclic antidepressant amitriptyline on morphine tolerance in rats. Male Wistar rats were implanted with two intrathecal (i.t.) catheters with or without a microdialysis probe, then received a continuous i.t. infusion of saline (control) or morphine (15 microg/h) and/or amitriptyline (15 microg/h) for 5 days. The results showed that amitriptyline alone did not produce an antinociceptive effect, while morphine alone induced antinociceptive tolerance and down-regulation of spinal glutamate transporters (GLAST, GLT-1, and EAAC1) in the rat spinal cord dorsal horn. Co-administration of amitriptyline with morphine attenuated morphine tolerance and up-regulated GLAST and GLT-1 expression. On day 5, morphine challenge (10 microg/10 microl) resulted in a significant increase in levels of the excitatory amino acids (EAAs), aspartate and glutamate, in CSF dialysates in morphine-tolerant rats. Amitriptyline co-infusion not only markedly suppressed this morphine-evoked EAA release, but also preserved the antinociceptive effect of acute morphine challenge at the end of infusion. Glial cells activation and increased cytokine expression (TNFalpha, IL-1beta, and IL-6) in the rat spinal cord were induced by the 5-day morphine infusion and these neuroimmune responses were also prevented by amitriptyline co-infusion. These results show that amitriptyline not only attenuates morphine tolerance, but also preserves its antinociceptive effect. The mechanisms involved may include: (a) inhibition of pro-inflammatory cytokine expression, (b) prevention of glutamate transporter down-regulation, and even up-regulation of glial GTs GLAST and GLT-1 expression, with (c) attenuation of morphine-evoked EAA release following continuous long term morphine infusion. PMID- 16697109 TI - Endogenous nociceptin/orphanin FQ signalling produces opposite spinal antinociceptive and supraspinal pronociceptive effects in the mouse formalin test: pharmacological and genetic evidences. AB - Nociceptin/orphanin FQ (N/OFQ) has been demonstrated to modulate nociceptive transmission via selective activation of N/OFQ peptide (NOP) receptors. Despite huge research efforts, the role(s) of the endogenous N/OFQ-NOP receptor system in pain processing remains incompletely understood. In the present study, we investigated the role of endogenous N/OFQ in the processing of tonic nociceptive input. To address this issue the effects of NOP-selective antagonists [Nphe1,Arg14,Lys15]N/OFQ-NH2 (UFP-101) and J-113397 on nociceptive behaviour, and the nociceptive phenotype of NOP receptor-deficient mice were tested in the mouse formalin test. Twenty microliters of 1.5% formalin solution was injected subcutaneously into the right hind paw causing a characteristic pattern of nociceptive behaviours (licking, biting and lifting of the injected paw). In control mice, the injection of formalin resulted in a classical biphasic nociceptive response with the first phase lasting from 0 to 10 min and the second phase from 15 to 45 min. UFP-101 at 10 nmol/mouse (but not at 1 nmol/mouse) produced antinociceptive action when injected intracerebroventricularly and a pronociceptive action when given intrathecally. Systemic administration of J 113397 (10 mg/kg, intravenously) and the genetic ablation of the NOP receptor gene both produced a significant increase of mouse nociceptive behaviour. Collectively, these results demonstrate that endogenous N/OFQ-NOP receptor signalling is activated during the mouse formalin test producing spinal antinociceptive and supraspinal pronociceptive effects. The overall effect of blocking NOP receptor signalling, by either systemic pharmacological antagonism or genetic ablation, indicates that the spinal antinociceptive action prevails over supraspinal pronociceptive effects. PMID- 16697110 TI - Quantitative sensory testing in the German Research Network on Neuropathic Pain (DFNS): standardized protocol and reference values. AB - The nationwide multicenter trials of the German Research Network on Neuropathic Pain (DFNS) aim to characterize the somatosensory phenotype of patients with neuropathic pain. For this purpose, we have implemented a standardized quantitative sensory testing (QST) protocol giving a complete profile for one region within 30 min. To judge plus or minus signs in patients we have now established age- and gender-matched absolute and relative QST reference values from 180 healthy subjects, assessed bilaterally over face, hand and foot. We determined thermal detection and pain thresholds including a test for paradoxical heat sensations, mechanical detection thresholds to von Frey filaments and a 64 Hz tuning fork, mechanical pain thresholds to pinprick stimuli and blunt pressure, stimulus/response-functions for pinprick and dynamic mechanical allodynia, and pain summation (wind-up ratio). QST parameters were region specific and age dependent. Pain thresholds were significantly lower in women than men. Detection thresholds were generally independent of gender. Reference data were normalized to the specific group means and variances (region, age, gender) by calculating z-scores. Due to confidence limits close to the respective limits of the possible data range, heat hypoalgesia, cold hypoalgesia, and mechanical hyperesthesia can hardly be diagnosed. Nevertheless, these parameters can be used for group comparisons. Sensitivity is enhanced by side-to-side comparisons by a factor ranging from 1.1 to 2.5. Relative comparisons across body regions do not offer advantages over absolute reference values. Application of this standardized QST protocol in patients and human surrogate models will allow to infer underlying mechanisms from somatosensory phenotypes. PMID- 16697111 TI - The efficacy of low level laser therapy for chronic neck pain. PMID- 16697112 TI - Quantitative Sensory Testing in children: practical considerations for research and clinical practice. PMID- 16697113 TI - Physical therapy and active exercises--an adequate treatment for prevention of late whiplash syndrome? Randomized controlled trial in 200 patients. AB - The aim of this study was to compare the effect of a physical therapy regimen including active exercises with the current standard treatment on reduction of pain 6 weeks and 6 months after whiplash injury caused by motor vehicle collision. Two hundred patients were enrolled in a prospective randomized controlled trial. In the standard group, treatment consisted of immobilization with a soft collar over 7 days. In the physical therapy group, patients were scheduled for 10 physical therapy appointments including active exercises within 14 days after enrollment. Pain intensity was rated by all patients daily during the first week, the sixth week, and 6 months after recruitment, using a numeric rating scale (0-10). Data analyses were performed by comparing the mean (over 1 week) pain scores between the two different treatment groups. Ninety-seven patients were randomly assigned to the standard treatment group and 103 to the physical therapy group. During the first week, there was no significant difference in mean pain intensity between the standard treatment group (4.76+/ 2.15) and the physical therapy group (4.36+/-2.14). However, after 6 weeks, mean pain intensity was significantly (p=0.002) lower in the physical therapy group (1.49+/-2.26 versus 2.7+/-2.78). Similarly, after 6 months, significantly (p<0.001) less pain was reported in the physical therapy group (1.17+/-2.13) than the standard treatment group (2.33+/-2.56). We conclude that a physical therapy regimen which includes active exercises is superior in reducing pain 6 weeks and 6 months after whiplash injury compared to the current standard treatment with a soft collar. PMID- 16697114 TI - Pharmacological characterization and autoradiographic distribution of alpha2 adrenoceptor antagonist [3H]RX 821002 binding sites in the chicken brain. AB - Knowledge about the noradrenergic system in birds is very scarce even though their biological diversity and complex social behavior make them an excellent model for studying neuronal functions and developmental biology. While the role of norepinephrine has been described in depth in a large number of central and peripheral functions in mammals, reports for avian species are limited. The radioligand [(3)H]RX 821002 ([(3)H]1,4-[6,7(n)3H]-benzodioxan-2-methoxy-2-yl)-2 imidazol) has been used to map and characterize alpha(2)-adrenoceptors through the chicken brain using in vitro autoradiography and membrane homogenates binding assays. [(3)H]RX 821002 showed a saturable and high affinity binding to a site compatible with alpha(2)-adrenoceptor, and to a serotonergic component. The autoradiographic assays displayed a similar alpha(2)-adrenoceptor distribution than those previously reported in birds using other radioligands such as [(3)H]UK 14304 ([(3)H]5-bromo-N-(4,5-dihydro-1H-imidazol-2-yl)-6-quinoxalinamine) or [(3)H]clonidine. [(3)H]RX 821002 binding pharmacological characterization was carried out in different chicken brain regions using membrane homogenates for competition assays with different alpha(2)-adrenoceptor agonists and antagonists drugs (oxymetazoline, BRL 44408 [2-(2H-(1-methyl-1,3-dihydroisoindole)methyl)-4,5 dihydroimidazole] ARC 239 [2-(2-4-(O-methoxyphenyl)-piperazin-1-yl)-ethyl-4,4 dimethyl-1,3-(2H,4H)-isoquinolindione], prazosin, UK 14304 and RX 821002). The results showed alpha(2A) as the predominant alpha(2)-adrenoceptor subtype in the chicken brain while alpha(2B)- and/or alpha(2C)-adrenoceptor subtypes were detected only in the telencephalon. RX 821002, serotonin (5-HT) and 8-OH-DPAT [8 hydroxy-2-(di-n-propylamino)tetralin] competition assays, and competition binding assays performed in the presence of serotonin demonstrated that [(3)H]RX 821002 binds with higher affinity to a serotonergic component, probably 5-HT(1A) receptors, than to the alpha(2)-adrenoceptors. Similar pharmacological properties for the alpha(2)-adrenoceptor component were observed both in rat and chicken brain. The results demonstrate that the different alpha(2)-adrenoceptor subtypes are present in chicken brain and suggest that these receptors are highly conserved through evolution. PMID- 16697116 TI - Hypoxia in presence of blockers of excitotoxicity induces a caspase-dependent neuronal necrosis. AB - When excitotoxic mechanisms are blocked, severe or prolonged hypoxia and hypoxia ischemia can still kill neurons, by a mechanism which is poorly understood. We studied this "non-excitotoxic hypoxic death" in primary cultures of rat dentate gyrus neurons. Many neurons subjected to hypoxia in the presence of blockers of ionotropic glutamate receptors developed the electron microscopic features of necrosis. They showed early mitochondrial swelling, loss of mitochondrial membrane potential and cytoplasmic release of cytochrome c, followed by activation of caspase-9, and by caspase-9-dependent activation of caspase-3. Caspase inhibitors were neuroprotective. These results suggest that "non excitotoxic hypoxic neuronal death" requires the activation in many neurons of a cell death program originating in mitochondria and leading to necrosis. PMID- 16697115 TI - Continuous exposure to glial cell line-derived neurotrophic factor to mature dopaminergic transplants impairs the graft's ability to improve spontaneous motor behavior in parkinsonian rats. AB - Functional recovery following intrastriatal transplantation of fetal dopaminergic neurons in animal models of Parkinson's disease is, at least in part, dependent on the number of surviving dopaminergic neurons and the degree of graft-derived dopaminergic reinnervation of the host striatum. In the present study, we analyzed whether continuous exposure of glial cell line-derived neurotrophic factor (GDNF) to mature dopaminergic grafts could further boost the functional outcome of widespread intrastriatal dopaminergic grafts. Rats with dopamine denervating lesions received multiple intrastriatal transplants of fetal dopaminergic cells and graft-induced behavioral effects were analyzed in drug induced and spontaneous motor behaviors. At three months after grafting, animals received intrastriatal injections of recombinant lentiviral vectors encoding for either human GDNF or the green fluorescent protein. Continuous exposure of GDNF to the grafts did not boost the functional recovery beyond what was observed in the control animals. Rather, in some of the spontaneous motor behaviors, animals in the GDNF-group showed deterioration as compared with control animals, and this negative effect of GDNF was associated with a down-regulation of the tyrosine hydroxylase enzyme. Based on these and our earlier results, we propose that intrastriatal administration of GDNF at the time of or shortly after grafting is highly effective in initially promoting the cell survival and fiber outgrowth from the grafts. However, once the grafts are mature, GDNF's ability to boost dopaminergic neurotransmission follows the same dynamics as for the native nigral dopaminergic neurons, which appears to be dependent on the concentration of GDNF. Since rather low doses of glial cell line-derived neurotrophic factor at nanogram levels appear to saturate these effects, it may be critical to adjust GDNF levels using tightly regulated gene expression systems. PMID- 16697117 TI - B-type natriuretic peptide is an independent predictor for cardiovascular death in patients with no clinical signs of ventricular dysfunction after acute myocardial infarction. AB - B-type natriuretic peptide (BNP) is released from cardiac ventricles in response to increased wall tension in patients with heart failure. It has been used as a biochemical marker for the diagnosis of congestive heart failure. BNP is also increased in patients with acute myocardial infarction, and is associated with an increased risk of cardiovascular mortality in those with impaired left ventricular function after myocardial infarction. We hypothesized that an increase in BNP soon after acute myocardial infarction is an independent predictor for long-term prognosis in patients with no clinical signs of left ventricular function. PMID- 16697118 TI - Atrial vulnerability is a major mechanism of paroxysmal atrial fibrillation in patients with Wolff-Parkinson-White syndrome. AB - Atrioventricular re-entrant tachycardia is the most common form of tachycardia in patients with Wolff-Parkinson-White (WPW) syndrome where one or more accessory pathways are present between atria and ventricles. Paroxysmal atrial fibrillation (PAF) is also a relatively common form of tachycardia in these patients. The mechanisms of PAF in patients with WPW syndrome are not completely understood. Previous studies have mainly focused on the role of accessory pathways in the development of PAF. However, up to 24% of patients still experience PAF after successful ablation of accessory pathways by surgical incision or radiofrequency catheter ablation techniques. We hypothesized that there are two primary mechanisms involved in the pathogenesis of PAF in patients with WPW syndrome. One is related to the accessory pathways that predispose the atria to fibrillation; the other is increased atrial vulnerability that is independent of the accessory pathways. Clarification of these mechanisms is critical in developing more effective means for the prevention or treatment of PAF in patients with WPW syndrome. PMID- 16697119 TI - Differential caspase-3 expression in noncancerous, premalignant and cancer tissues of stomach and its clinical implication. AB - BACKGROUND: Caspase-3 is a critical apoptosis-promoting element but its status during stepwise gastrocarcinogenesis needs to be further clarified. MATERIALS AND METHODS: By the use of frozen tissue microarrays constructed with the tissue spots cored from defined histological regions in tissue blocks, the pattern of caspase-3 expression in noncancerous, premalignant (atrophic gastritis and intestinal metaplasia) tissue and cancer spots were analyzed under the same experimental conditions by the methods of immunohistochemistry and mRNA-in situ hybridization. RESULTS: Caspase-3 was expressed in all 34 of the noncancerous mucosa (100%), in 16 of the 17 premalignant tissues (94.1%) and in 15 of the 48 gastric cancers (31.3%). The incidences of caspase-3 detection were significantly different (p<0.01) between noncancerous mucosa and intestinal as well as diffuse gastric cancers. CONCLUSION: Down-regulated caspase-3 is closely correlated with gastric cancer formation and would be a potential indicator of tumor formation and progression. Helicobacter pylori (H. pylori; Hp) infection is but not the only one element responsible to the enhanced caspase-3 expression in gastric epithelia. PMID- 16697120 TI - [Microarray-based comparative genomic hybridization in the study of constitutional chromosomal abnormalities]. AB - Chromosomal aberrations are the first cause of mental impairment and dysmorphism. Rearrangements involving large chromosomal segments can be detected by standard chromosome analysis using GTG-banding, but this technique is not suited for the detection of small chromosome abnormalities. Array comparative genomic hybridisation (array-CGH) is a method used to detect segmental DNA copy number alterations. Recently, advances in this technology have enabled high-resolution examination for identifying genetic alterations and copy number variations on a genome-wide scale. This review describes the current genomic array platforms and CGH methodologies and highlights their applications for studying constitutional disease. PMID- 16697121 TI - [Human cytogenetics. From 1956 to 2006]. AB - The correct enumeration of human chromosomes, only established in 1956, has marked the starting point of the modern cytogenetics. The introduction of banding techniques, then of in situ hybridization techniques, and now of genomic microarray technology allowed a dramatic development of cytogenetics of which the main applications to basic and medical research are evoked in this review. PMID- 16697122 TI - Trisomy 8 as the sole chromosomal aberration in acute myeloid leukemia and myelodysplastic syndromes. AB - Trisomy 8 as the sole abnormality is the most common karyotypic finding in acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS), occurring in approximately 5% and 10% of the cytogenetically abnormal cases, respectively. However, despite the high frequency of +8, much remains to be elucidated as regards its epidemiology, etiology, clinical impact, association with other chromosomal abnormalities, cell of origin, and functional and pathogenetic consequences. Here, we summarize and review these various aspects of trisomy 8, focusing on AMLs and MDS harboring this abnormality as a single change. PMID- 16697123 TI - Interleukin-2 receptor beta chain locus rearrangement in a T-cell acute lymphoblastic leukemia. AB - A translocation t(1;22)(p13;q13) was detected in a child with T-cell acute lymphoblastic leukemia (T-ALL). FISH studies showed that the breakpoint was located in the 5' part of the interleukin-2 receptor beta chain (IL2RB) locus, but could only be located distal to 1p13.3 on the partner chromosome. This is the first case of the IL2RB locus rearrangement in T-ALL. The localization of the breakpoint suggests that the chromosomal translocation results in deregulation of IL2RB expression. PMID- 16697124 TI - A double-blind, placebo-controlled trial of amantadine, propranolol, and their combination for the treatment of cocaine dependence in patients with severe cocaine withdrawal symptoms. AB - BACKGROUND: This trial evaluated the efficacy of amantadine, propranolol and their combination in cocaine dependent patients with severe cocaine withdrawal symptoms. METHODS: Cocaine withdrawal symptom severity was measured by the cocaine selective severity assessment (CSSA). One hundred and ninety-nine patients with high scores on the CSSA participated in a 10-week double-blind trial. Patients were randomly assigned to receive amantadine (300 mg/day), propranolol (100mg/day), a combination of amantadine (300 mg/day) and propranolol (100mg/day) or matching placebo capsules. The primary outcome measure was cocaine abstinence. RESULTS: In the intent-to-treat sample, there were no significant differences between the four medication groups in treatment retention. The odds of cocaine abstinence showed a marginally significant increase over time in the propranolol group (p=0.06) but not in the other three groups. In highly medication-adherent patients, treatment retention was significantly better in the propranolol group compared to the placebo group (p=0.01) and the odds of cocaine abstinence increased significantly over time in the propranolol group but not in the other three groups. CONCLUSION: In the intent-to-treat sample, none of the three active treatments (propranolol, amantadine or their combination) was significantly more effective than placebo in promoting abstinence from cocaine among patients who entered treatment with more severe cocaine withdrawal symptoms. Among patients highly adherent to study medication, propranolol treatment was associated with better treatment retention and higher rates of cocaine abstinence compared to placebo. PMID- 16697125 TI - Behavioral and neurochemical interactions between Group 1 mGluR antagonists and ethanol: potential insight into their anti-addictive properties. AB - Blockade of the mGluR5 subtype of Group 1 metabotropic glutamate receptor (mGluRs) reduces the rewarding effects of ethanol (EtOH), while the effects of mGluR1a blockade remain under-investigated. The present study compared the effects of pretreatment with the mGluR5 antagonist MPEP and the mGluR1a antagonist CPCCPOEt upon behavioral and neurochemical variables associated with EtOH reward in alcohol-preferring C57BL/6J mice. Pretreatment with either antagonist (0-10 mg/kg, IP) dose-dependently reduced measures of EtOH reward in an operant self-administration paradigm and the maximally effective antagonist dose (10 mg/kg) also blocked the expression of EtOH-induced place conditioning, as well as EtOH consumption under 24-h free-access conditions. MPEP pretreatment did not significantly alter the EtOH dose-locomotor response function; however, it prevented EtOH-induced changes in extracellular dopamine, glutamate and GABA in the nucleus accumbens (NAC). In contrast, CPCCOEt shifted the EtOH dose response function downwards, enhanced the capacity of higher EtOH doses to elevate NAC levels of GABA and lowered extracellular dopamine and glutamate below baseline following EtOH injection. It is suggested that the "anti-alcohol" effects of MPEP may involve an attenuation of the neurochemical signals mediating EtOH reward, whereas those of CPCCOEt may involve an increased sensitivity to the inhibitory effects of EtOH upon brain and behavior. PMID- 16697126 TI - Evaluation of antiviral activity of phenolic compounds and derivatives against rabies virus. AB - Human rabies is a viral disease with a great impact on public health, mainly on account of its fatal course in the majority of cases. Despite the well established prophylaxis by immunization, rabies is believed to be responsible for 40,000-70,000 human deaths per year, mostly in endemic areas. Palliative support and experimental protocols to avoid death have been employed with no expressive results, with the exception of a recent human case of recovery from rabies. No antiviral drugs are currently available to fight against this infection. In combination with the prophylaxis, an antiviral drug would be useful for human rabies treatment, providing enhanced protection against the encephalitis caused by the virus. Phenolic compounds are derived from the secondary plant metabolism, although they can also be obtained by synthetic processes. Many studies have shown a great range of pharmacological effects for these substances, including vasodilatation, antiallergenic, antiinflammatory and antiviral properties, among others. In this study, the potential in-vitro anti-rabies activity of 24 synthetic phenolic compounds was evaluated using McCoy cells and PV rabies strain. The cytotoxicity (CC50) was assayed by the MTT method and the antiviral activity (IC50) was estimated by the inhibition of viral cytopathic effects. Isoprinosine and ketamine were used as positive controls. The tested compounds showed selectivity indices (SI=CC50/IC50) ranging from 1.0 to 3.9. Six phenolic compounds failed to inhibit the cytopathic effect to any degree, and four showed SI > or = 3.0. According to these results, some probable structure-activity relationships are suggested. It was observed that the presence of free hydroxyl and ether groups influenced the anti-rabies activity. However, additional studies are required to establish these relationships. PMID- 16697127 TI - The occurrence and characterization of Campylobacter jejuni and C. coli in organic pigs and their outdoor environment. AB - The occurrence and species distribution of thermophilic Campylobacter was investigated in organic outdoor pigs. An increased exposure of outdoor pigs to C. jejuni from the environment may cause a shift from a normal dominance of C. coli to more C. jejuni, which may imply a concern of reduced food safety. Bacteriological methods for determination of Campylobacter excretion level were combined with colony-blot hybridization and real-time PCR for specific detection of C. jejuni in pigs. Campylobacter was isolated from pigs (n=47), paddock environment (n=126) and wildlife (n=44), identified to species by real-time PCR and sub-typed by serotyping (Penner) and pulse-field gel electrophoresis (PFGE) genotyping. All pigs excreted Campylobacter (10(3)-10(7) CFU g(-1) faeces) from the age of 8-13-weeks old. C. jejuni was found in 29% of pigs in three consecutive trials and always in minority to C. coli (0.3-46%). C. jejuni and C. coli were isolated from 10% and 29% of the environmental samples, respectively, while crow-birds and rats harboured C. jejuni. Individual pigs hosted several strains (up to nine serotypes). The paddock environment was contaminated with C. coli serotypes similar to pig isolates, while most of the C. jejuni serotypes differed. C. jejuni isolates of different origin comprised few similar serotypes, just one identical genotype was common between pigs, environment and birds. In conclusion, the occurrence of C. jejuni varied considerably between the three groups of outdoor pigs. Furthermore, transfer of C. jejuni to the outdoor pigs from the nearby environment was not predominant according to the subtype dissimilarities of the obtained isolates. PMID- 16697128 TI - Quantitative relationship of dioxin-responsive gene expression to dioxin response element in Hep3B and HepG2 human hepatocarcinoma cell lines. AB - Dioxin response element (DRE) is a cis-acting DNA sequence mediating the 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD)-induced gene expression. The present study was undertaken to elucidate TCDD-responsive gene expression profiles and their relationships to the number of DREs in liver cancer cells. Hep3B and HepG2 human hepatocarcinoma cells were exposed to 50-nM TCDD for 0, 1, 2 and 4h in culture, after which gene expression profiles were analyzed by the microarray hybridization using a chip containing 24,000 cDNAs prepared from the human liver. The TCDD-responsive expression levels in each gene were calculated by dividing the densitometric values of the hybridization signal for h1, h2 and h4 by that of h0, followed by transformation of the resulting data into a log scale with the base of 2. Up- and down-regulated gene expressions were defined as >0.585 and < 0.585 by the log scale (>1.5 and <1/1.5 arithmetically), respectively, exhibited at any time after h0. Hep3B and HepG2 cells had 27 and 58 TCDD-responsive, up regulated genes, respectively, of which 78% (21/27) and 62% (36/58) had one or more DREs. Of these 85, 80 genes were up-regulated exclusively in one of the two lines, with CYP1A1 and PPP1R15A being so regulated in both lines. Expression levels of the up-regulated genes at h1, h2 and h4 were correlated with each other (P<0.01) and the mean of these regressed to the number of DRE(s) in both lines (P<0.01). However, expression of a total of 93 TCDD-responsive, down-regulated genes, of which 46% contained DRE(s), had no relation to the number of DRE(s). In conclusion, results suggest that DREs may cooperatively mediate the expression of TCDD-responsive genes in liver cancer cells. PMID- 16697129 TI - The immune system as a target for environmental chemicals: Xenoestrogens and other compounds. AB - The immune system in higher organisms is under integrated control and has the capacity to rapidly respond to the environment. Recently, there has been a significant increase in the prevalence of allergic diseases. Environmental factors likely play a major role in the explosion of allergy. Although the "hygiene hypothesis" may explain the increase in allergic diseases which are prone to T helper 2 (Th2) immune responses, recent findings highlight the possible involvement of environmental xenobiotic chemicals which can modulate normal immune function. Interestingly, several reports suggest that the prevalence of systemic lupus erythematosus, a Th2-type autoimmune disease, is also increasing, although the development of high-sensitivity immunological tests may be a possible cause. The increased prevalence of autoimmune disease in women, the sexual dimorphism of the immune response, and the immunomodulatory effects of sex steroids, have focused attention on the role of chemicals which influence sex steroids in the development of immune diseases. Moreover, recent reports indicate that some environmental chemicals can work on nuclear hormone receptors, other than sex hormone receptors, and modulate immune reactions. This review focuses on the impact of environmental chemicals on immune system function and pathogenesis of immune diseases, including allergy and autoimmune diseases. PMID- 16697130 TI - Non-surgical deep intra-uterine transfer of in vitro produced porcine embryos derived from sex-sorted frozen-thawed boar sperm. AB - Embryos and offspring of a pre-determined sex have been produced in pigs using AI and IVF with unfrozen sperm, and after surgical insemination with sex-sorted frozen-thawed sperm. The aims of this study were to demonstrate that sex-sorted frozen-thawed boar sperm could be incorporated into pig IVF for the production of embryos of a pre-determined sex and that these embryos could be successfully non surgically transferred. Oocytes were matured in vitro, fertilised with either unsorted or sex-sorted frozen-thawed sperm and cultured until the eight-cell stage. These embryos were then transferred to recipients (n = 7) non-surgically (n = 70 embryos per sow). Oocyte cleavage was similar between sex-sorted (1538/5044; 30.5%) and unsorted (216/756; 28.6%) frozen-thawed sperm, and PCR sex determination of the embryos confirmed that they were of the predicted sex (n = 16). Delayed return to oestrus (>23 days) was observed in five recipient sows (71.4%). Fetal sacs were observed by transcutaneous ultrasound on Day 18 in one of these sows. Pre-sexed porcine IVP embryos can be successfully produced using sex-sorted frozen-thawed boar sperm, and these embryos are capable of initiating pregnancies when transferred to recipients. However, further refinement of porcine ET protocols are required to enable development to term. PMID- 16697132 TI - Ketogenic diet for treatment of infantile spasms. AB - This study sought to evaluate the efficacy, tolerability, and safety of a ketogenic diet (KD) in the treatment of infantile spasms (IS), incorporating a revised protocol based on our previous experience with KD. We undertook a retrospective analysis of 43 children who suffered from catastrophic IS and tried KD from June 1995 to May 2004 in two Korean epilepsy centers. Outcome measures included seizure frequency, electroencephalograms (EEGs), adverse reactions, and neurological development. Overall, the diet achieved the seizure-free state in 53.5% (23/43) of patients and a greater than 90% reduction of seizure frequency in 62.8% (27/43) of patients. The seizure outcomes were highly concordant with improvements in EEG findings and development. In addition, KD could be maintained more safely and more comfortably because of our revised protocol that included an initial non-fasting diet, a short-term trial of 8 months, a more protein-rich diet with a lipid to non-lipid ratio of 3:1, and liquid ketogenic milk. Most complications were transient and KD was well tolerated in most cases. KD can be an effective alternative therapy for catastrophic IS, and additional benefits may be derived from constant revision of the diet in the future. PMID- 16697133 TI - [Characteristics of newly managed HIV-infected patients: hospital Saint-Antoine, Paris 2002-2003]. AB - OBJECTIVES: The authors had for aim to assess incident HIV+ patients in Saint Antoine hospital, Paris in 2002-2003 (transfers excluded). METHODS: Sociodemographic, clinical, and virological data were collected to compare French and sub-Saharan patients. RESULTS: Three hundred new HIV+ patients were identified: mean age 36.2 year; ratio M/W 65/35; origin: Caucasian 43%, African 44%, Maghrebian 8%, Asian 3%; nationality: French 45%, sub-Saharan African 44%; illegal aliens 65%; employed 51%; transmission: heterosexual 54%, homosexual 39%, intravenous drug user (IVDU) 2%; circumstances for diagnosis: HIV exposure 34%, primary infection 9%, symptoms/AIDS 23%, pregnancy 6%, other 28%; CDC stage: A 77%, B 9%, C 14%; mean T-CD4+ 374/mm3, median HIV-RNA 30780 cp/mL; co-infection HBV 7.3% HCV 5%. OUTCOME: 88% still followed up in Saint-Antoine, 2% transferred, 9% lost to follow-up, 1% dead. A significant difference was shown: 1) between French (N = 123) and African (N = 46) men for heterosexual transmission (10 vs 91%), working status (85 vs 28%), T-CD4+ (354 vs 251/mm3), outcome (lost to follow-up 5 vs 24%) [P < 0.01]; 2) between French (N=13) and African (N = 85) women for age (41 vs 32 years), working status (38 vs 10%), stage A (54 vs 81%) [P < 0.05]; 3) between African patients according to sex (46M/85W) for age (39 vs 32 years), working status (28 vs 10%; P = 0.01), outcome (lost to follow-up 24 vs 6%) [P < 0.01]. CONCLUSION: This study highlights the persistence of HIV in native homosexual French men and the increasing prevalence in African migrant with precarious social status. PMID- 16697134 TI - Identification and characterization of an intervening sequence within the 23S ribosomal RNA genes of Edwardsiella ictaluri. AB - Comparison of the 23S rRNA gene sequences of Edwardsiella tarda and Edw. ictaluri confirmed a close phylogenetic relationship between these two fish pathogen species and a distant relation with the 'core' members of the Enterobacteriaceae family. Analysis of the rrl gene for 23S rRNA in Edw. ictaluri revealed the presence of an intervening sequence (IVS) in helix-45. This new 98bp IVS shared 97% nucleotide identity with Salmonella typhimurium helix-45 IVS. Edw. ictaluri helix-45 IVS was present in all Edw. ictaluri strains analyzed and in at least six rrl operons within each cell. Fragmentation of 23S rRNA due to IVS excision by RNase III was observed by methylene blue staining of ribosomal RNA extracted from Edw. ictaluri isolates. This is the first report of an IVS in the 23S rRNA gene of the genus Edwardsiella. PMID- 16697135 TI - Microbial community analysis of a biogas-producing completely stirred tank reactor fed continuously with fodder beet silage as mono-substrate. AB - The bioconversion of renewable raw material to biogas by anaerobic microbial fermentation processes in completely stirred tank reactors (CSTR) is a valuable alternative resource of energy especially for rural areas. However, knowledge about the microorganisms involved in the degradation of plant biomass is still poor. In this study, a first analysis of the biogas-forming process within a CSTR fed continuously with fodder beet silage as mono-substrate is presented in the context of molecular data on the microbial community composition. As indicated by the conventional process parameters like pH value, content of volatile fatty acids, N:P ratio and the biogas yield, the biogas-forming process within the CSTR occurred with a stable and efficient performance. The average biogas yield based on volatile solids was 0.87m(3)kg(-1) at an organic loading rate of 1.2-2.3kgm( 3)d(-1). This amounts to 94% of the theoretical maximum. In order to identify microorganisms within the CSTR, a 16S rDNA clone library was constructed by PCR amplification applying a prokaryote-specific primer set. One hundred and forty seven clones were obtained and subsequently characterized by amplified rDNA restriction analysis (ARDRA). The sequences of 60 unique ARDRA patterns were estimated in a length of approximately 800-900bp each. Four of them were assigned to the domain Archaea and 56 to the domain Bacteria. Within the domain Archaea, all clones showed a close relationship to methanogenic species. Major bacterial groups represented in the clone library were the class Clostridia of the phylum Firmicutes (22% of all 16S rDNA clones), the class Deltaproteobacteria of the phylum Proteobacteria (24%), the class Bacilli of the phylum Firmicutes (22%) and members of the phylum Bacteroidetes (21%). Within these major groups, the highest biodiversity was found within the class Clostridia (35% of all operational taxonomic units). Members of the phyla Actinobacteria and Spirochaetes were represented only by 5 and 2 clonal sequences, respectively. PMID- 16697136 TI - Molecular cloning and tissue expression of chicken AdipoR1 and AdipoR2 complementary deoxyribonucleic acids. AB - AdipoR1 and AdipoR2 belong to a novel class of transmembrane receptors that mediate the effects of adiponectin. We have cloned the chicken AdipoR1 and AdipoR2 complementary deoxyribonucleic acids (cDNA) and determined their expression in various tissues. We also investigated the effect of feed deprivation on the expression of AdipoR1 or AdipoR2 mRNA in the chicken diencephalon, liver, anterior pituitary gland, and adipose tissue. The chicken AdipoR1 and AdipoR2 cDNA sequences were 76-83% identical to the respective mammalian sequences. A hydrophobicity analysis of the deduced amino acid sequences of chicken AdipoR1/AdipoR2 revealed seven distinct hydrophobic regions representing seven transmembrane domains. By RT-PCR, we detected AdipoR1 and AdipoR2 mRNA in adipose tissue, liver, anterior pituitary gland, diencephalon, skeletal muscle, kidney, spleen, ovary, and blood. AdipoR1 or AdipoR2 mRNA expression in various tissues was quantified by real-time quantitative PCR, and AdipoR1 mRNA expression was the highest in skeletal muscle, adipose tissue and diencephalon, followed by kidney, ovary, liver, anterior pituitary gland, and spleen. AdipoR2 mRNA expression was the highest in adipose tissue followed by skeletal muscle, liver, ovary, diencephalon, anterior pituitary gland, kidney, and spleen. We also found that a 48 h feed deprivation significantly decreased AdipoR1 mRNA quantity in the chicken pituitary gland, while AdipoR2 mRNA quantity was significantly increased in adipose tissue (P<0.05). We conclude that the AdipoR1 and AdipoR2 genes are ubiquitously expressed in chicken tissues and that their expression is altered by feed deprivation in the anterior pituitary gland and adipose tissue. PMID- 16697137 TI - Non-ventilation during early incubation in combination with dexamethasone administration during late incubation: 1. Effects on physiological hormone levels, incubation duration and hatching events. AB - This study investigated the effect of non-ventilation of the incubator during the first 10 days of incubation and its combination with dexamethasone administration at day 16 or 18 of incubation on hatching parameters and embryo and post-hatch chick juvenile physiology. A total of 2400 hatching eggs produced by Cobb broiler breeders were used for the study. Blood samples were collected at day 18 of incubation, at internal pipping stage (IP), at the end of hatch (day-old chick) and at 7-day-post-hatch for T(3), T(4) and corticosterone levels determination. From 448 to 506 h of incubation, the eggs were checked individually in the hatcher every 2h for pipping and hatching. The results indicate that non ventilation during the first 10-day shortened incubation duration up to IP, external pipping (EP) and hatch, had no effect on hatchability and led to higher T(3) levels at IP but lower corticosterone levels at 7-day-post-hatch. The injection of dexamethasone at days 16 and 18 of incubation affected hatching and blood parameters in both the ventilated and non-ventilated embryos differentially and the effect was dependent on the age of the embryo. Dexamethasone increased T(3) levels and T(3)/T(4) ratios but the effect was greater with early non ventilation of eggs. Dexamethasone decreased hatchability but the effect was greater when injected at day 16 and especially in ventilated embryos. The effects of incubation protocols and dexamethasone treatments during incubation were still apparent in the hatched chicks until 7 days of age. The changes in T(3), T(4) and corticosterone levels observed in response to the early incubation conditions and late dexamethasone treatments in this study suggest that incubator ventilation or non-ventilation may influence the hypothalamic-pituitary-adrenal axis (HPA) regulation of stress levels (in terms of plasma corticosterone levels) and thyroid function in the embryo with impact on incubation duration, hatching events and early post-hatch life of the chick. Our results also suggest that some stages of development are more sensitive to dexamethasone administration as effects can be influenced by early incubation protocols. PMID- 16697138 TI - [Management of ventilator acquired pneumonia]. AB - Ventilator-associated pneumonia occurs in the evolution of 8 to 70% of patients in the Intensive Care Unit. It is the main site of nosocomial infection for mechanically ventilated patients. Nosocomial pneumonia represents an important cause of morbidity and mortality, despite progresses in antibiotic prescription, use of intensive care and prevention. This review is based on the ATS guidelines, and reviews epidemiology, diagnosis and treatment of ventilator-acquired pneumonia, in non-immunocompromised adults. PMID- 16697139 TI - Conservation patterns in different functional sequence categories of divergent Drosophila species. AB - We have explored the distributions of fully conserved ungapped blocks in genome wide pair-wise alignments of recently completed species of Drosophila: D. melanogaster, D. yakuba, D. ananassae, D. pseudoobscura, D. virilis, and D. mojavensis. Based on these distributions we have found that nearly every functional sequence category possesses its own distinctive conservation pattern, sometimes independent of the overall sequence conservation level. In the coding and regulatory regions, the ungapped blocks were longer than in introns, UTRs, and nonfunctional sequences. At the same time, the blocks in the coding regions carried a 3N + 2 signature characteristic of synonymous substitutions in the third-codon position. Larger block sizes in transcription regulatory regions can be explained by the presence of conserved arrays of binding sites for transcription factors. We also have shown that the longest ungapped blocks, or "ultraconserved" sequences, are associated with specific gene groups, including those encoding ion channels and components of the cytoskeleton. We discuss how restraining conservation patterns may help in mapping functional sequence categories and improve genome annotation. PMID- 16697140 TI - Biomechanical responses of human temporomandibular joint disc under tension and compression. AB - The purpose of this study was to observe the microscopic changes in the temporomandibular joint (TMJ) disc in response to tension and compression, and to study the mechanism of disc failure when subjected to mechanical stresses. The scanning electron microscope was used to observe topographic characteristics of TMJ disc samples obtained from 13 cadavers before and during biomechanical experiments. The stress relaxation experiments were conducted at different strain levels (2, 3, 4 and 6%). The uniaxial tensile experiments were carried out at a constant strain rate (0.05 mm/s). The confined compression tests were performed with 3 different indenters (2, 3 and 4mm in diameter) for 150 s and 3 h. The maximal tensile strain of the disc was 5% (nearly equal to 0.22 MPa of tensile stress) in the mediolaterally tensile direction. Typical wavelike structure of the collagen fibrils of the disc was present at 2-4% strain ranges. Tensile and shear damage to local collagen fibrils was observed in specimens of the intermediate zone and the posterior band at 6% strain level. Changes in the collagen network from a wavelike structure to distortion observed on the surface of the testing samples were reversible in the 150-s indentation, but severe, irreversible breakdown and deformation of the collagen-proteoglycan network occurred in those specimens that had been compressed for 3h. Persistent and prolonged compression inevitably resulted in irrecoverable disc failure. Mechanical stress is a crucial factor in breakdown of the TMJ disc. PMID- 16697141 TI - Mineralization at the interface of implants. AB - Osseointegration of implants is crucial for the long-term success of oral implants. Mineralization of the bone's extracellular matrix as the ultimate step of a mature bone formation is closely related to implant osseointegration. Osteogenesis at oral implants is a complex process, driven by cellular and acellular phenomena. The biological process of the maintenance and emergence of minerals in the vicinity of oral implants is influenced to a great extent by biophysical parameters. Implant-related structural and functional factors, as well as patient-specific factors, govern the features of osteogenesis. To understand the influence of these factors in peri-implant bone mineralization, it is important to consider the basic biological processes. Biological and crystallographic investigations have to be applied to evaluate mineralization at implant surfaces at the different hierarchical levels of analysis. This review gives insight into the complex theme of mineral formation around implants. Special focus is given to new developments in implant design and loading protocols aimed at accelerating osseointegration of dental implants. PMID- 16697142 TI - A 2-stage procedure combining maxillary advancement by distraction technique with mandibular setback surgery in patients with cleft lip and palate. AB - A 2-stage procedure combining maxillary advancement by distraction technique with mandibular setback surgery was used to correct jaw deformities in 5 patients with severe maxillary retrusion secondary to cleft lip and palate. First, a Le Fort I maxillary osteotomy was performed. Immediately after maxillary distraction, the distraction device was removed. The advanced maxilla was fixed with miniplates after adjusting the length and direction of advancement, and mandibular setback surgery was performed simultaneously to obtain a normal occlusal relationship. This 2-stage procedure resulted in stable occlusion and a markedly improved facial profile. PMID- 16697143 TI - Spontaneous sensory recovery in non-innervated radial forearm flaps used for head and neck reconstruction. AB - Although several studies have reported the use of reinnervated microvascular free flaps for oro-pharyngeal reconstruction, it has been known for some time that non innervated flaps demonstrate spontaneous sensory recovery. This study sought to evaluate the degree of such spontaneous recovery in 50 radial forearm flaps used for mucosal reconstruction of head and neck ablative defects. The recovery of sensation to pinprick, light touch and temperature was tested a mean of 38 months (range 15-71) after surgical insetting. Two-point discrimination was also sought. Although 18 flaps (36%) remained anaesthetic, partial recovery in one or more modalities was present in 28 patients (56%). A recovery in all modalities of sensation in at least two-thirds of the flap area was recorded in 4 patients (8%). The mean 2-point static discrimination for fascio-cutaneous flaps was 18.9mm. PMID- 16697144 TI - Functional outcomes after organ preservation treatment in head and neck cancer: a critical review of the literature. AB - The use of radiation therapy and/or chemotherapy in advanced head and neck cancer is increasing in popularity, driven by the notion that sparing the organs of speech and swallowing from surgical resection will also spare function. This critical review of the literature considered functional outcomes after organ preservation to assess the impact of such treatment on speech, swallowing and quality of life in patients with head and neck cancer. Literature searches were conducted on several library databases. A total of 50 relevant articles were identified and found to meet the inclusion criteria specified a priori. The majority of reports suggested that organ preservation techniques have the potential to result in swallowing disorders, often related to dysmotility of the oropharyngeal and laryngeal structures, and resulting in frequent episodes of aspiration. This may lead to the need for enteral feeding in the short term for some patients while, in others, this need is life long. Speech does not appear to be affected to the same degree as swallowing. These results suggest that organ preservation does not translate into function preservation for all patients with head and neck cancer. PMID- 16697145 TI - Effects of irradiation on bone remodelling around mandibular implants: an experimental study in dogs. AB - This research focuses on the effects of radiotherapy on bone remodelling around mandibular implants in dogs. After bilateral extraction of the mandibular premolars and first 2 molars, each of 11 beagles received 8 mandibular implants. Four animals were irradiated 4 weeks after implantation and 4 others 8 weeks before implantation; the remaining 3 did not receive radiotherapy. Irradiation consisted of 10 daily fractions of 4.3Gy (60)Co. Fluorochromes were given at implantation and irradiation to allow the measurement of bone apposition. The dogs were killed 6 months after implantation. Each hemi-mandible was processed according to bone-specific histological techniques. New bone formation was visible around 85 of the 88 implants. Stimulated mandibular remodelling was attested in both irradiated groups by increased porosity and numerous labelled osteons. Resorption was more pronounced in the group irradiated after implantation, but osteon formation appeared unvarying. Osseointegration was thus shown to be compatible with bone irradiation as bone turnover activities were maintained throughout the experiment. As the apposition stage of the remodelling cycle appears crucial to achieve optimal osseointegration, its normal completion should be taken into account in clinical practice by respecting a 6-month period between irradiation and implantation. PMID- 16697146 TI - Effect of postoperative radiotherapy on the functional result of implants placed during ablative surgery for oral cancer. AB - The purpose of this retrospective study was to evaluate the survival of dental implants placed during ablative surgery in the interforaminal region of the original edentulous mandible in patients with squamous cell carcinoma of the oral cavity in relation to postoperative radiotherapy. Forty-eight patients treated in 1996-2003 with surgery alone or in combination with postoperative radiotherapy were analysed. In all patients, 2 to 4 Branemark Mk II/III 2-phase implants were placed during tumour resection. A total of 139 implants were placed of which 61 (21 patients) received postoperative radiotherapy: 60-68 Gy as a boost dose on the primary tumour site and 10-68 Gy on the symphyseal area. No difference was found in percentage of functional dentures on implants between the radiated and non-radiated groups. The success rate of osseointegration was 97% in the postoperative irradiated group and 100% in the non-irradiated group. The prosthetic success rate (75%) was lower because in 12 of the 48 patients (34 implants) a functional denture could not be fitted due to tumour recurrence or metastasis (7 patients, 22 implants) or for psychological reasons (4 patients, 12 implants), independent of whether radiotherapy was administered. Postoperative radiotherapy does not affect the osseointegration of dental implants placed during tumour ablation and the ultimate number of functional dentures. Primary implant placement in edentulous mandibles may have advantages over secondary implant placement in patients with oral squamous cell carcinoma. PMID- 16697147 TI - Vertical distraction of fibula transplant in a case of mandibular defect caused by shotgun injury. AB - This case demonstrates the successful aesthetic and functional reconstruction of a complex facial gun-shot injury with extended bone defects and soft tissue destructions using a 3-step procedure. Initially, a reconstruction plate was inserted, later a fibula transplant enabled the basic reconstruction and finally was distructed in a 3rd session. The rationale behind the sequencing of surgical sessions was the extended bony defect and soft-tissue destruction. The main problem in this type of wound is hypoxia or anoxia of the receptor bed for the transplant. A microvascular anastomosized bone transplant is necessary for sufficient oxygen tension in the recipient site. The anatomical dimensional disproportion of the transplanted free fibula graft and the shape of the mandible were corrected prior to the insertion of dental implants by means of vertical distraction. PMID- 16697148 TI - A feasibility study of computer-assisted health-related quality of life data collection in patients with oral and oropharyngeal cancer. AB - Touchscreen technology (TST) has been widely used in the field of medicine. The purpose of this study was to pilot a standard TST system to administer the University of Washington Quality of Life Questionnaire (UWQOL) in a routine head and neck oncology clinic. The UWQOLv4 was completed using TST and a semi structured interview was used to evaluate patients' experience of the data collection process. Forty-one of 44 consecutive cancer patients agreed to participate in the study. Although over three-quarters of patients reported never having used a computer, all but one found the UWQOLv4 'easy' or 'very easy' to complete using the TST. They preferred the TST method to paper copy and the vast majority completed the TST in 10min or less. TST provides a very suitable mechanism for routine health-related quality of life data collection. The system is easy to programme and relatively inexpensive. PMID- 16697149 TI - A simple method for bone positioning of mandibular segments. AB - A simple bone-positioning technique is reported, using a combination of bone plates and a flexible tube filled with acrylic polymer. The flexible tube is solidified by injection of acrylic monomer. This technique is applicable for bone positioning after mandibular resection as well as positioning of the proximal segment after sagittal split ramus osteotomy. This technique has the advantages of greater simplicity and flexibility compared to previously reported methods of bone positioning. PMID- 16697150 TI - In vivo selection of Enterobacter aerogenes with reduced susceptibility to cefepime and carbapenems associated with decreased expression of a 40 kDa outer membrane protein and hyperproduction of AmpC beta-lactamase. AB - The mechanism(s) of resistance or decreased susceptibility to cefepime (FEP) and/or imipenem (IMP) in three consecutive isolates of Enterobacter aerogenes (Ea1, Ea2 and Ea3) cultured from bronchial aspirates of the same patient after treatment with ceftriaxone and FEP were studied. Identification was performed with the VITEK 2 system. All three isolates showed identical pulsed-field gel electrophoresis patterns and were resistant (minimum inhibitory concentrations (MICs)) to cefoxitin (MIC, >1024 mg/L), cefotaxime (CTX; MIC, 32-128 mg/L) and ceftazidime (CAZ; MIC, 32-128 mg/L) but susceptible to meropenem (MIC, Glu. Three outer membrane proteins (OMPs) of 51 kDa, 40 kDa and 38 kDa were observed in the three isolates by sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE) (10% polyacrylamide gels with 4 M urea), although expression of the 40 kDa OMP was reduced in Ea2. In conclusion, decreased susceptibility to FEP and IMP in Ea2 is related to reduced expression of a 40 kDa OMP and hyperproduction of AmpC, whereas resistance to FEP in Ea3 is associated with hyperproduction of an altered AmpC. PMID- 16697151 TI - A naturalistic multicenter study of intramuscular olanzapine in the treatment of acutely agitated manic or schizophrenic patients. AB - BACKGROUND: We conducted a naturalistic, multicenter, 24-hour, nonrandomized, observational study describing for the first time the effectiveness and safety of intramuscular (IM) olanzapine to control agitation and aggression in "real world" patients with psychosis. The data thus obtained was compared with that reported from randomized double-blind clinical trials. METHOD: 92 patients attending psychiatric emergency settings were enrolled. The study subjects were 44 male and 48 female patients with a mean age of 36.5+/-12 years and DSM-IV-TR diagnoses of schizophrenia (48.9%), psychotic disorder not specified (23.9%) or bipolar disorder (27.2%). 10 mg IM olanzapine was administered to all patients. An optional second injection was permitted> or =2 hours later in line with hospital policy. Evaluations (PANSS-EC and CGI-S) were performed at baseline and 2 and 24 hours following the IM injection. RESULTS: Two hours after IM olanzapine was administered, a mean decrease of -9.6 in the PANSS-EC from a baseline score of 26.5 was recorded. At the 24-hour endpoint a statistically and clinically significant reduction in the PANSS-EC scores (11.6+/-5.3) was observed as compared with values at study entry (26.5+/-5.9) and at 2 hours endpoint (16.9+/ 9.3), which represent a mean decrease of -14.9 and -5.3, respectively. CONCLUSION: The present naturalistic study provides naturalistic data on the effectiveness of IM olanzapine in the treatment of acute agitation in patients with schizophrenia or bipolar mania that is in line the data obtained in randomized double-blind clinical trials. PMID- 16697152 TI - The factor structure of the General Health Questionnaire (GHQ): a scaled version for general practice in Spain. AB - BACKGROUND: The 28-item version of the General Health Questionnaire (GHQ-28) developed by Goldberg and Hillier in 1979 is constructed on the basis of a principal components analysis of the GHQ-60. When used on a Spanish population, a translation of the GHQ-28 developed for an English population may lead to worse predictive values. METHODS: We used our Spanish sample to replicate the entire process of construction of the GHQ-28 administered in a primary-care setting. RESULTS: Two shorter versions were proposed: one with six scales and 30 items, and the other with four scales and 28 items. CONCLUSIONS: The resulting GHQ-28 was a successful adaptation for use on the Spanish sample. When compared with the original version, only 21 items were the same. Moreover, contrary to the English version, which groups sleep problems and anxiety in the same scale, a scale with items related exclusively to 'Sleep disturbances' was found. PMID- 16697153 TI - Duloxetine in the treatment of major depressive disorder: a placebo- and paroxetine-controlled trial. AB - OBJECTIVE: Duloxetine doses of 80 and 120 mg/day were assessed for efficacy and safety in the treatment of major depressive disorder (MDD). METHODS: In this randomized, double-blind trial, patients age > or =18 meeting DSM-IV criteria for MDD were randomized to placebo (N=99), duloxetine 80 mg/day (N=93), duloxetine 120 mg/day (N=103), or paroxetine 20 mg/day (N=97). The primary outcome measure was mean change from baseline in the 17-item Hamilton rating scale for depression (HAMD(17)) total score after 8 weeks of treatment; a number of secondary efficacy measures also were assessed. Safety and tolerability were assessed via collection and analysis of treatment-emergent adverse events (TEAEs), vital signs, and weight. The Arizona sexual experiences scale was used to assess sexual functioning. Patients who had a > or =30% reduction from baseline in the HAMD(17) total score at the end of the acute phase entered a 6-month continuation phase where they remained on the same treatment as they had taken during the acute phase; efficacy and safety/tolerability outcomes were assessed during continuation treatment. RESULTS: More than 87% of patients completed the acute phase in each treatment group. Duloxetine-treated patients (both doses) showed significantly greater improvement (P<0.05) in the HAMD(17) total score at week 8 compared with placebo. Paroxetine was not significantly different from placebo (P=0.089) on mean change on the HAMD(17). Duloxetine 120 mg/day also showed significant improvement on most secondary efficacy measures (six of nine) compared with placebo while duloxetine 80 mg/day (three of nine) and paroxetine (three of nine) were significantly superior to placebo on fewer secondary measures. HAMD(17) mean change data from this study and an identical sister study were pooled as defined a priori for the purposes of performing a non-inferiority test versus paroxetine. Both duloxetine doses met statistical criteria for non inferiority to paroxetine. TEAE reporting rates were low in all treatment groups and no deaths occurred in the acute or continuation phases. CONCLUSIONS: The efficacy of duloxetine at doses of 80 and 120 mg/day in the treatment of MDD was demonstrated. Tolerability, as measured by TEAEs, and safety were similar to paroxetine 20 mg/day and consistent with previous published data on duloxetine in the treatment of MDD. PMID- 16697154 TI - Barotropic phase transition between the lamellar liquid crystal phase and the inverted hexagonal phase of dioleoylphosphatidylethanolamine. AB - The phase transition between the lamellar liquid crystal (Lalpha) phase and the inverted hexagonal (H(II)) phase of dioleoylphosphatidylethanolamine (DOPE) in aqueous NaCl solutions was observed by means of differential scanning calorimetry (DSC) under ambient pressure and light-transmittance technique under high pressure. The pressure dependence of the transition temperature (dT/dp) and the thermodynamic quantities for the Lalpha/H(II) transition were compared with those of another phase transition found in the DOPE bilayer membrane, which is the transition from the lamellar crystal (Lc) phase to the Lalpha phase. The dT/dp value of the Lalpha/H(II) transition was about 3.5 times as large as that of the Lc/Lalpha transition while the thermodynamic quantities were significantly smaller than those of the latter to the contrary. Comparing the enthalpy and volume behavior of the Lalpha/H(II) transition with that of the Lc/Lalpha transition, we concluded that the Lalpha/H(II) transition can be regarded as the volume-controlled transition for the reconstruction of molecular packing. PMID- 16697155 TI - Boosting support vector regression in QSAR studies of bioactivities of chemical compounds. AB - In this paper, boosting has been coupled with SVR to develop a new method, boosting support vector regression (BSVR). BSVR is implemented by firstly constructing a series of SVR models on the various weighted versions of the original training set and then combining the predictions from the constructed SVR models to obtain integrative results by weighted median. The proposed BSVR algorithm has been used to predict toxicities of nitrobenzenes and inhibitory potency of 1-phenyl[2H]-tetrahydro-triazine-3-one analogues as inhibitors of 5 lipoxygenase. As comparisons to this method, the multiple linear regression (MLR) and conventional support vector regression (SVR) have also been investigated. Experimental results have shown that the introduction of boosting drastically enhances the generalization performance of individual SVR model and BSVR is a well-performing technique in QSAR studies superior to multiple linear regression. PMID- 16697156 TI - [Screening of congenital dislocation of the hip in the newborn]. PMID- 16697157 TI - [Pediatrics and dental emergency]. PMID- 16697158 TI - [Ultrasonography for hip joint dislocation in neonates]. PMID- 16697159 TI - [Allegation of sexual abuses during conflictual divorces]. PMID- 16697160 TI - [Animal dander allergy in children]. PMID- 16697161 TI - [Common classification of medical acts in clinical pediatrics]. PMID- 16697162 TI - [Vaccinal status in children with seizures: a retrospective study conducted in the Bordeaux child hospital]. AB - Vaccination is a common act in medicine. Some serious side effects are always feared in a preventive action, mainly among high-risk patients such as epileptic children or children having already experienced a seizure. OBJECTIVES: To study consequences of such background on the vaccine medical practice. POPULATION AND METHODS: A retrospective study comparing the vaccine statute of children with or without case history of seizures was carried out by the neurologic and paediatric emergencies departments. The study compared 55 with seizures versus 109 without. RESULTS: On the whole, the 2 groups were insufficiently vaccinated. A statistically significant difference was highlighted between the 2 groups for the vaccination coverage by vaccine DTP (diphtheria-tetanus-pertussis) (P=0.017) and MMR (measles-mumps-rubella) (P=0.004). However, concerning the vaccination against hepatitis B, no difference was found. CONCLUSION: The usual contra indications of these vaccines do not explain this difference and progress must be made to improve the vaccination coverage of epileptic children. PMID- 16697163 TI - [Approach of the general practitioner to the adolescent and third party]. PMID- 16697164 TI - [Fetal nephropathies]. PMID- 16697165 TI - [Role of parents for adolescent's outpatient clinic]. PMID- 16697166 TI - [Intravenous analgesia for pain control in paediatric lower extremity surgery]. PMID- 16697167 TI - [Zoonoses in childhood and companion animals]. PMID- 16697168 TI - [RePOP, a network for prevention and management of obesity in children in Lyon area]. PMID- 16697169 TI - Pharmacoscintigraphic evaluation of lipid dry powder budesonide formulations for inhalation. AB - Lung deposition of new formulations of budesonide, using solid lipid microparticles (SLmP) as a pharmaceutically acceptable filler and carrier for inhalation aerosols, and administered from a dry powder inhaler (Cyclohaler), were compared with that from Pulmicort Turbuhaler. Six healthy volunteers took part in a three-way randomized cross-over study, and inhaled a nominal dose of 400 microg budesonide, labelled with 99mTc, on each study day. Lung deposition was determined by gamma scintigraphy and by a pharmacokinetic method. The percentage of dose (SD) in the whole lung was 49.9 (3.7)% for the lipidic matricial form (M) and 62.8 (4.9)% for the lipidic physical blend formulation (PB). These results corresponded well with the in vitro fine particle assessment. In comparison with data recorded in literature for in vivo deposition obtained with Pulmicort Turbuhaler, it was estimated that lung deposition was 1.5 and 2.0 times higher for the M and PB formulations, respectively. Furthermore, the relative drug availability obtained from the pharmacokinetic evaluation, expressed as the percentage of pulmonary absorption of the comparator product, was 154% and 220% for M and PB, respectively. The results of the present study indicate that pulmonary administration using SLmP gives a prominent and significant increase in budesonide lung deposition. PMID- 16697170 TI - The use of agar as a novel filler for monolithic matrices produced using hot melt extrusion. AB - The use of filler materials in an extended release monolithic polymer matrix can lead to a vastly altered release profile for the active pharmaceutical ingredient. A range of excipients for use in monolithic matrices have been discussed in the literature. The body of work described in this research paper outlines the use of agar as a novel filler material in a hot melt extruded polymer matrix. Several batches of matrix material were prepared with Diclofenac sodium used as the active pharmaceutical ingredient (API). Agar and microcrystalline cellulose were used as the filler materials in varying ratios, to examine the effect of % filler content as well as filler type on the properties of the hot melt extruded matrix. The resultant extrudates were characterised using steady state parallel plate rheometry, differential scanning calorimetry (DSC) and dissolution testing. The rheometry analysis concluded that the fillers used resulted in an increase in the matrix viscosity. The DSC scans obtained showed negligible effects on the melting behavior of the matrix as a result of the filler inclusion. Dissolution analysis showed that the presence of the fillers resulted in a slower release rate of API than for the matrix alone. The results detailed within this paper indicate that agar is a viable filler for extended release hot melt produced dosage forms. PMID- 16697171 TI - Compaction behaviour and new predictive approach to the compressibility of binary mixtures of pharmaceutical excipients. AB - The compressibility of three pharmaceutical excipients (microcrystalline cellulose, lactose and anhydrous calcium phosphate) and their binary mixtures was studied. The aim of this work was to observe the impact of the mass composition of the mixture on the compressibility. The single-compound materials and their mixtures were compacted using instrumented presses. It allowed obtaining compression cycles (i.e., force-displacement curves) which were associated with energy measurements (specific compaction energy, Esp cp and specific expansion energy, Esp exp). It was observed that for the mixtures studied, the change of Esp cp with the mass composition could be fitted using a linear relationship (it was not the case with Esp exp). A linear relationship between the porosity of mixture's compacts and the mass composition was also obtained. Heckel's plots were then obtained for the three excipients and the mixtures. The mean yield pressure was calculated with the "in-die-method" and the "out-of-die method". A proportional relationship was not valid for the mean yield pressures. But, a predictive approach was proposed in order to obtain indirectly the mean yield pressure of a binary mixture if the data of the single materials were known. It used the linear mixing rule observed with the porosity. The validity was verified and compared with the experimental values. This comparison showed that it was possible to predict the mean yield pressure of binary mixtures from the accessible data of the single excipients. PMID- 16697172 TI - Versatility of biodegradable poly(D,L-lactic-co-glycolic acid) microspheres for plasmid DNA delivery. AB - In this study, we have optimized different formulations of DNA encapsulated into PLGA microspheres by correlating the protocol of preparation and the molecular weight and composition of the polymer, with the main characteristics of these systems in order to design an efficient non-viral gene delivery vector. For that, we prepared poly(D,L-lactic-co-glycolic acid) (PLGA) microparticles with an optimized water-oil-water double emulsion process, by using several types of polymers (RG502, RG503, RG504, RG502H and RG752), and characterized in terms of size, zeta potential, encapsulation efficiency (EE%), morphology, DNA conformation, release kinetics, plasmid integrity and erosion. The size of the particles ranged between 0.7 and 5.7 microm depending on the protocol of formulation and the molecular mass of the polymer used. The microspheres prepared by using in their formulation polymers of high molecular weight (RG503 and RG504) were bigger in size than in the case of using a lower molecular weight polymer (RG502). The EE (%) of plasmid DNA increased with increasing the molecular mass of the polymer and by using the most hydrophilic polymer RG502H, which contains terminal acidic groups in its structure. The plasmid could be encapsulated without compromising its structural and functional integrity. Also a protective effect of PLGA on endonuclease digestion is observed. Plasmid DNA release from microspheres composed of low molecular weight or hydrophilic polymers, like RG502H, was faster than from particles containing high molecular weight or hydrophobic polymers. These PLGA microspheres could be an alternative to the viral vectors used in gene therapy, given that may be used to deliver genes and other bioactive molecules, either very rapidly or in a controlled manner. PMID- 16697173 TI - Accumulation of copper by Acremonium pinkertoniae, a fungus isolated from industrial wastes. AB - Acremonium pinkertoniae isolated from zinc wastes was studied to understand the mechanisms that allow living organisms to thrive in polluted environments and the possible role of the fungus in the redistribution and cycling of copper. The fungus was cultured on solid media supplemented with copper sulfate at increasing concentrations. At high doses it was observed that the mycelia acquired a characteristic blue color. This was accompanied by morphological changes and the formation of crystalloid structures within thickened cell walls. The material was further analysed with EDX, X-ray powder diffraction and IR spectroscopy. It was demonstrated that A. pinkertoniae is able to accumulate over 20% dry weight of copper by what probably is a chitin-glucan complex. PMID- 16697174 TI - Inactivation of polygalacturonase and pectate lyase produced by pH tolerant fungus Fusarium moniliforme NCIM 1276 in a liquid medium and in the host tissue. AB - Fusarium moniliforme NCIM 1276 produced pH dependent an extracellular polygalacturonase (PG) and pectate lyase (PL) at pH 5 and pH 8, respectively. In the extracellular medium about 20.3% PG and 54% of PL protein concentrations were present in the active state at pH 5 and pH 8, respectively, whereas in intracellularly, more than 86% of both protein contents remained in the active state at all pH tested. We found two possible reasons, end-product inhibition and effect of environmental pH on conformation of the proteins after their release into the medium. Additionally, in infected tomato and cauliflower plants, the fungus secreted similar proteins which were located near to the epidermal and vascular regions of the hypocotyls. In infected tissues, between 26.9% and to 41.5% of PG and only 0.84%-13.4% of PL protein concentrations were present in active state. Thus, the medium/cell sap pH and concentrations of substrate/end products seem to play an important role in fungal invasion during plant pathogenesis are discussed with current literature. PMID- 16697175 TI - Cyclic compressive mechanical stimulation induces sequential catabolic and anabolic gene changes in chondrocytes resulting in increased extracellular matrix accumulation. AB - Overcoming the limited ability of articular cartilage to self-repair may be possible through tissue engineering. However, bioengineered cartilage formed using current methods does not match the physical properties of native cartilage. In previous studies we demonstrated that mechanical stimulation improved cartilage tissue formation. This study examines the mechanisms by which this occurs. Application of uniaxial, cyclic compression (1 kPa, 1 Hz, 30 min) significantly increased matrix metalloprotease (MMP)-3 and MMP-13 gene expression at 2 h compared to unstimulated cells. These returned to constitutive levels by 6 h. Increased MMP-13 protein levels, both pro- and active forms, were detected at 6 h and these decreased by 24 h. This was associated with tissue degradation as more proteoglycans and collagen had been released into the culture media at 6 h when compared to the unstimulated cells. This catabolic change was followed by a significant increase in type II collagen and aggrecan gene expression at 12 h post-stimulation and increased synthesis and accumulation of these matrix molecules at 24 h. Mechanical stimulation activated the MAP kinase pathway as there was increased phosphorylation of ERK1/2 and JNK as well as increased AP-1 binding. Mechanical stimulation in the presence of the JNK inhibitor, SP600125, blocked AP-1 binding preventing the increased gene expression of MMP-3 and -13 at 2 h and type II collagen and aggrecan at 12 h as well as the increased matrix synthesis and accumulation. Given the sequence of changes, cyclic compressive loading appears to initiate a remodelling effect involving MAPK and AP-1 signalling resulting in improved in vitro formation of cartilage. PMID- 16697176 TI - Caesium sorption by hydrated cement as a function of degradation state: experiments and modelling. AB - To provide reliable K(d) data for Cs required for the performance assessment of cement-based radioactive waste repositories, two complementary approaches were followed. First, Cs sorption was determined on a range of hydrated cement paste (HCP) and mortar samples of CEM I and CEM V for different degradation states and solution compositions, as well as on some single mineral phases. Second, a surface complexation-diffuse layer model previously developed by Pointeau et al. [Pointeau, I., Marmier, N., Fromage, F., Fedoroff, M., Giffaut, E., 2001. Cs and Pb uptake by CSH phases of hydrated cement. Material Research Society Symposium Proceedings, 663, 105-113] for Cs sorption on synthetic CSH phases was simplified to facilitate its application to whole HCP and mortars or concrete, following re assessment of the model parameters. All measurements were compared with model predictions. The sorption data obtained on the different solid phases as a function of conditions corroborate that CSH minerals are the main sorbing phase for Cs in HCP. The data also clearly show the important influence of pH and the dissolved concentration of Na, K and Ca on K(d). It is further suggested that a decrease of pH is concomitant with a decrease of the Ca/Si ratio and a corresponding increase in surface sites with high affinity for Cs and, thus, K(d). Elevated concentrations of cations able to compete with Cs for these sites lead to a decrease of K(d), on the other hand. The simplified model was applied to the sorption measurements performed within this study as well as to a variety of literature data, mainly K(d) values for a variety of fresh HCP and mortar or concrete samples based on different samples of Ordinary Portland Cement as well as blended cements. The results show that the model can be applied reasonably well to a very large variety of conditions in terms of solid and solution compositions that cover a range of K(d) values from 10(-4) to ca. 3.2m(3)/kg. The large scatter typically observed for Cs sorption, especially on fresh HCP samples prepared from different formulations, can be explained quantitatively by the variable concentrations of Na and K in the respective solutions, which compete with Cs for fixation sites. On the other hand, the comparatively uniform conditions in degraded HCP typically render the prediction of K(d) values less uncertain than in case of fresh HCP. PMID- 16697177 TI - Evolution of iron speciation during hydration of C4AF. AB - It is now well accepted and demonstrated that calcium silicate, calcium aluminate and calcium sulfo aluminate (ettringite, AFm) phases exhibit a good capability to fix metals and metalloids. Unfortunately the role of minor phases and especially calcium-ferric aluminate phase, shorthand C4AF is not well defined. In other systems like in soils or sediments iron phases play a key role in the fixation of pollutant. In cement sorption isotherms, indicated that various metals can be retained by the C4AF hydrated products. Therefore the capabilities of those phase to retain heavy metal should not be neglected. Previous investigations have shown that the minerals formed during the hydration of C4AF are similar to those formed from C3A (pure tri-calcium aluminate) under comparable conditions. Nevertheless no investigation was conducted at the molecular level and there is still a controversy whether Fe substitutes for Al in the hydrated minerals in whole or in part, or if it forms FeOOH clusters scattered throughout the matrix. In this context we have conducted XAS experiments using synchrotron radiation. It was found that the hydration of C4AF forms C3AH6 (hydrogarnet) in which Fe randomly substitutes for Al as well as an amorphous FeOOH phase. Intermediate products like AFm (i.e., an ill organized lamellar phase) are also formed but rapidly evolve to C3AH6; iron does not seem to be incorporated in the AFm structure. PMID- 16697178 TI - Harnessing microbial activities for environmental cleanup. AB - Human activities have released large amounts of toxic organic and inorganic chemicals into the environment. Toxic waste streams threaten dwindling drinking water supplies and impact terrestrial, estuarine and marine ecosystems. Cleanup is technically challenging and the costs based on traditional technologies are exceeding the economic capabilities of even the richest countries. Recent advances in our understanding of the microbiology contributing to contaminant transformation and detoxification has led to successful field demonstrations. Hence, harnessing the activity of naturally occurring bacteria, particularly the power of anaerobic reductive processes, is a promising approach to restore contaminated subsurface environments, protect drinking water reservoirs and to safeguard ecosystem health. PMID- 16697179 TI - RNA splicing: group I intron crystal structures reveal the basis of splice site selection and metal ion catalysis. AB - The group I intron has served as a model for RNA catalysis since its discovery 25 years ago. Four recently determined high-resolution crystal structures complement extensive biochemical studies on this system. Structures of the Azoarcus, Tetrahymena and bacteriophage Twort group I introns mimic different states of the splicing or ribozyme reaction pathway and provide information on splice site selection and metal ion catalysis. The 5'-splice site is selected by formation of a conserved G.U wobble pair between the 5'-exon terminus and the intron. The 3' splice site is identified through stacking of three base triples, in which the middle triple contains the conserved terminal nucleotide of the intron, OmegaG. The structures support a two-metal-ion mechanism for group I intron splicing that might have corollaries to group II intron and pre-mRNA splicing by the spliceosome. PMID- 16697180 TI - Mammary stem and progenitor cells: tumour precursors? AB - Several groups have proposed that mammary epithelial cell (MEC) populations, in common with other epithelia, have stem and progenitor sub-populations that are long-lived and provide most of the growth potential during ductal (and perhaps lobuloalveolar) outgrowth. In this review, we describe what is known about normal development, particularly with respect to the growth potential and regenerative capacity of mouse MEC populations. We have developed a theoretical model in order to understand how the activity of the somatic stem/progenitor cell compartment during mammary gland development could affect the demographic of adult MEC populations. This demographic is likely to be key to understanding tumour risk, since long-lived cells provide great advantages in the process of cancer development. PMID- 16697181 TI - Microencapsulation of Zanthoxylum limonella oil (ZLO) in glutaraldehyde crosslinked gelatin for mosquito repellent application. AB - Glutaraldehyde (GA) crosslinked gelatin (G) microcapsules containing Zanthoxylum limonella oil (ZLO) were prepared by coacervation technique. The effect of various parameters such as variation of oil-loading, gelatin concentration and degree of crosslinking on release rate of oil were studied. Scanning electron microscopy (SEM) was used to understand the surface characteristics of microcapsules. FTIR-results indicated the absence of any significant interaction between polymer and oil. PMID- 16697182 TI - Induction of pullulanase production in Bacillus cereus FDTA-13. AB - Studies were carried out on the production of pullulanase by a newly isolated strain Bacillus cereus FDTA-13. High titres of the enzyme were obtained in a medium containing branched polysaccharides. To further enhance the yield, induction of pullulanase using conventional inducers were studied. Maltooligosaccharides (maltose to maltotetraose) when added in the medium individually, or in a 1:1 combination of maltotriose and maltotetraose resulted different levels of pullulanase compared to control. Growth under carbon limited conditions (5 g l(-1)) with inducers resulted remarkably enhanced pullulanase activity. Pullulanase activity was severely repressed in presence of glucose. Low levels of pullulanase was observed in nitrogen limited medium, even with combinations of several maltosaccharides. PMID- 16697183 TI - Automatic control of biomass gasifiers using fuzzy inference systems. AB - A fuzzy controller for biomass gasifiers is proposed. Although fuzzy inference systems do not need models to be tuned, a plant model is proposed which has turned out very useful to prove different combinations of membership functions and rules in the proposed fuzzy control. The global control scheme is shown, including the elements to generate the set points for the process variables automatically. There, the type of biomass and its moisture content are the only data which need to be introduced to the controller by a human operator at the beginning of operation to make it work autonomously. The advantages and good performance of the fuzzy controller with the automatic generation of set points, compared to controllers utilising fixed parameters, are demonstrated. PMID- 16697184 TI - Comparison of activated carbon and bottom ash for removal of reactive dye from aqueous solution. AB - The adsorption of reactive dye from synthetic aqueous solution onto granular activated carbon (GAC) and coal-based bottom ash (CBBA) were studied under the same experimental conditions. As an alternative to GAC, CBBA was used as adsorbent for dye removal from aqueous solution. The amount of Vertigo Navy Marine (VNM) adsorbed onto CBBA was lower compared with GAC at equilibrium and dye adsorption capacity increased from 0.71 to 3.82 mg g(-1), and 0.73 to 6.35 mg g(-1) with the initial concentration of dye from 25 to 300 mg l(-1), respectively. The initial dye uptake of CBBA was not so rapid as in the case of GAC and the dye uptake was slow and gradually attained equilibrium. PMID- 16697185 TI - Response surface optimization of the critical medium components for carbonyl reductase production by Candida viswanathii MTCC 5158. AB - Culture conditions were optimized for the growth and carbonyl reductase production by a novel yeast strain Candida viswanathii. Response surface methodology was applied for the critical medium components (initial pH, mannitol, yeast extract and calcium chloride) identified earlier by one-factor-at-a-time approach. Central composite design was used for the optimization studies. Using this methodology, the optimal values for the concentration of mannitol, initial pH, yeast extract and calcium chloride were 1.9, 7.5, 1.6 and 4, respectively. This medium was projected to produce, theoretically, growth having an optical density of 1.1 (600 nm) and an enzyme activity of 81.5 U/ml. Using this optimized medium, an experimental growth of 1.1 OD (600 nm) and enzyme activity 80.9 U/ml verified the applied methodology. This approach for medium optimization led to an enhancement of the growth and enzyme activity by 1.3 and 2.3 times higher, respectively, as compared to the unoptimized media. PMID- 16697186 TI - Privileged structure based ligands for melanocortin receptors--4,4-disubstituted piperidine derivatives. AB - Homologation and cyclization back to the chiral methine of compound 3 yields achiral 4,4-disubstituted piperidine privileged structures (e.g., 8a) useful in the construction of melanocortin 4 receptor (MC4R) ligands. The piperidine nitrogen was replaced with carbon, oxygen, sulfur, and sulfone with minor erosion of binding. The methyl cyclohexane substituent was the most potent while significant affinity was still seen for smaller lipophilic groups such as ethyl. PMID- 16697187 TI - Design, synthesis, and SAR studies on a series of 2-pyridinylpiperazines as potent antagonists of the melanocortin-4 receptor. AB - A series of 2-pyridinylpiperazines derived from beta-Ala-(2,4-Cl)Phe dipeptide was synthesized for the study of their SARs and possible interactions with the MC4 receptor. Compounds such as 11k (Ki=6.5 nM) possessed high potency. PMID- 16697188 TI - Synthesis of [11C]Iressa as a new potential PET cancer imaging agent for epidermal growth factor receptor tyrosine kinase. AB - Iressa (Gefitinib) is an orally active inhibitor of epidermal growth factor receptor tyrosine kinase (EGFR-TK) involved in cell signal transduction processes critical to proliferation, apoptosis, repair, and angiogenesis of cancer cells. [11C]Iressa was first designed and synthesized as a new potential positron emission tomography (PET) cancer imaging agent for EGFR-TK in 30-40% radiochemical yield with 4.0-6.0 Ci/micromol specific activity at end of bombardment (EOB). PMID- 16697189 TI - Discovery of 3-arylpropionic acids as potent agonists of sphingosine-1-phosphate receptor-1 (S1P1) with high selectivity against all other known S1P receptor subtypes. AB - A series of 3-arylpropionic acids were synthesized as S1P1 receptor agonists. Structure-activity relationship studies on the pendant phenyl ring revealed several structural features offering selectivity of S1P1 binding against S1P2-5. These highly selective S1P1 agonists induced peripheral blood lymphocyte lowering in mice and one of them was found to be efficacious in a rat skin transplantation model, supporting that S1P1 agonism is primarily responsible for the immunosuppressive efficacy observed in preclinical animal models. PMID- 16697190 TI - Potent hFPRL1 (ALXR) agonists as potential anti-inflammatory agents. AB - We report the discovery of potent agonists for the human formyl-peptide-like 1 receptor (hFPRL1). These compounds did not act at a closely related receptor denoted human formyl peptide receptor (hFPR) up to 10 microM concentration. Recent studies have indicated that agonizing this receptor may promote resolution of inflammation. In an exploratory study, a novel hFPRL1 agonist showed efficacy in a mouse ear inflammation model following oral administration. PMID- 16697191 TI - Neurotrophic peptide aldehydes: solid phase synthesis of fellutamide B and a simplified analog. AB - A combination of solid phase and solution phase synthetic methods have been used to complete the total synthesis of the neurotrophic lipopeptide aldehyde fellutamide B (2). The beta-hydroxy aliphatic tail was prepared by regioselective reductive opening of a cyclic sulfate, and later coupled to a solid phase resin. The synthetic compound was then examined in cytotoxicity and nerve growth factor (NGF) induction assays. A simplified analog of fellutamide B also showed activity. PMID- 16697192 TI - New pyrrolopyrimidin-6-yl benzenesulfonamides: potent A2B adenosine receptor antagonists. AB - A new series of 4-(1,3-dialkyl-2,4-dioxo-2,3,4,5-tetrahydro-1H-pyrrolo[3,2 d]pyrimidin-6-yl)benzenesulfonamides has been identified as potent A2B adenosine receptor antagonists. The products have been evaluated for their binding affinities for the human A2B, A1 and A3 adenosine receptors. 6-(4-{[4-(4 Bromobenzyl)piperazin-1-yl]sulfonyl}phenyl)-1,3-dimethyl-1H-pyrrolo[3,2 d]pyrimidine-2,4(3H,5H)-dione (16) showed a high affinity for the A2B adenosine receptor (IC50=1 nM) and selectivity (A1: 183x; A3: 12660x). Synthesis and SAR of this novel class of compounds showing improved absorption properties is presented herein. PMID- 16697193 TI - Synthesis and structure-activity relationships of 3,4-diaminocyclobut-3-ene-1,2 dione CXCR2 antagonists. AB - A novel series of 3,4-diaminocyclobut-3-ene-1,2-diones was prepared and found to show potent inhibitory activity of CXCR2 binding and IL-8-mediated chemotaxis of a CXCR2-expressing cell line. Microsome stability and Caco2 studies were subsequently used to show that compounds of this chemotype are predicted to have good oral bioavailability and are thus suitable for pharmaceutical development. PMID- 16697194 TI - Design and synthesis of orally active pyrrolidin-2-one-based factor Xa inhibitors. AB - A series of novel, non-basic 3-(6-chloronaphth-2-ylsulfonyl)aminopyrrolidin-2-one based factor Xa (fXa) inhibitors, incorporating an alanylamide P4 group, was designed and synthesised. Within this series, the N-2-(morpholin-4-yl)-2-oxoethyl derivative 24 was shown to be a potent, selective fXa inhibitor with good anticoagulant activity. Moreover, 24 possessed highly encouraging rat and dog pharmacokinetic profiles with excellent oral bioavailabilities in both species. PMID- 16697195 TI - Crystal structure, spectroscopic, and biological study of the copper(II) complex with third-generation quinolone antibiotic sparfloxacin. AB - The neutral mononuclear copper(II) complex with the quinolone antibacterial drug sparfloxacin has been prepared and characterized with IR, UV-vis, and EPR spectroscopies and X-ray crystallography. The interaction of the complex with calf-thymus DNA has also been investigated and the antimicrobial activity has been evaluated against three different microorganisms. PMID- 16697196 TI - Structure-activity relationships of 1,5-biaryl pyrroles as EP1 receptor antagonists. AB - The preliminary SAR of a series of novel 1,5-biaryl pyrrole EP1 receptor antagonists derived from compound 1 is described. Replacement of the benzyl group of 1 with isosteric groups was investigated. The most effective replacement was found to be the isobutyl group. The cyclopentylmethyl and cyclohexylmethyl groups were also effective benzyl replacements. The cyclohexylmethyl derivative 19 demonstrated the lowest metabolic clearance within this series. In addition, several high affinity substituted benzyl analogues were also identified. Compound 39 was found to have good bioavailability in rats and demonstrated efficacy in the established FCA preclinical model of inflammatory pain with a calculated ED50 of 9.2mg/kg. PMID- 16697197 TI - Lamin A/C assembly defects in Emery-Dreifuss muscular dystrophy can be regulated by culture medium composition. AB - Emery-Dreifuss muscular dystrophy results from mutations in either emerin or lamin A/C and is caused by loss of some unknown function of emerin-lamin A/C complexes. This function must be of special importance in the skeletal and cardiac muscles that are affected by the disease. Some lamin A/C mutant proteins form 'nuclear foci' in the nucleoplasm when overexpressed by transient transfection and similar aggregates have been seen in cultured skin fibroblasts from patients with Emery-Dreifuss muscular dystrophy, suggesting that mis assembly of the A-type lamina may be involved in the pathogenesis. Whereas an earlier study of cultured skin fibroblasts compared several different missense mutations in lamin A/C, we have chosen to study one particular Emery-Dreifuss mutation (R249Q) in greater detail. We found that the proportion of fibroblast nuclei containing abnormal lamin A/C aggregates can vary from 0.5 to 23.6% depending on the culture conditions. In particular, switching from a 'slow growth' medium to 'rapid growth' media increased both the number and size of nuclear aggregates. Similar results were obtained with fibroblasts from a second unrelated patient with the same mutation. In contrast to these aggregates of endogenous lamin A/C, 'nuclear foci' formed after transfection of mouse embryo fibroblasts by mutant lamin A/C were not affected by culture conditions. Faulty assembly of the nuclear lamina by mutated lamin A/C molecules could be partly responsible for the disease phenotype, though this has not been proven. The present study suggests that inappropriate lamin A/C assembly may be preventable by manipulation of cell growth conditions. PMID- 16697198 TI - An unusual pathologic feature associated with dermatomyositis. AB - We present a case of juvenile dermatomyositis with unusual histopathologic findings. The child presented with a course consistent with dermatomyositis, a diagnosis confirmed by finding reticulotubular aggregates in endothelial cells on electron microscopy. However, histopathology of his muscle biopsy revealed a striking pattern of glycogen accumulation, to an extent similar to that seen in glycogen storage diseases; this degree of accumulation could potentially confound histopathologic diagnosis. PMID- 16697199 TI - Glucocorticoid receptor isoforms generate transcription specificity. AB - Glucocorticoids are necessary for life and are essential in all aspects of health and disease as they regulate processes from mitosis to apoptosis, from metabolism to growth and development. However, responses to glucocorticoids vary among individuals, cells and tissues. Recent evidence indicates that multiple glucocorticoid receptor (GR) isoforms are generated from one single GR gene by alternative splicing and alternative translation initiation. These isoforms all have unique tissue distribution patterns and transcriptional regulatory profiles. Furthermore, each is subject to various post-translational modifications that affect receptor function. Thus, increasing evidence suggests that unique GR isoform compositions within cells could determine the cell-specific response to glucocorticoids. Here, we discuss a new molecular model potentially underlying tissue-specific glucocorticoid resistance and selectivity. PMID- 16697200 TI - Amphibian muscle regeneration--dedifferentiation or satellite cells? AB - Until recently, the cell biology of mammalian muscle repair following damage appeared to be completely different from the formation of new muscles in regenerated appendages of Amphibia. Mammalian muscle repair occurs through the mobilization of muscle satellite cells, whereas the new muscle in amphibian appendage regeneration was believed to arise by dedifferentiation of myofibres to form myoblasts. But recent work shows that muscle satellite cells are also involved in amphibian regeneration and the controversy about the reality of muscle dedifferentiation is heating up again. PMID- 16697201 TI - Does postural chain muscular stiffness reduce postural steadiness in a sitting posture? AB - This study investigated the effect of postural chain muscular stiffening on postural steadiness when it is rhythmically perturbed by respiration. It consisted of an analysis of centre of pressure (CP) displacements when constant sub-maximum pushing efforts were performed in a sitting posture. Muscular stiffness, assessed by surface electromyography (iEMG), was imposed at two controlled levels, using two intensities of pushing effort (20% and 40% of the maximum voluntary contraction: 20MVC and 40MVC). Lumbo-pelvic mobility was varied using two different support areas at the seat contact (100% and 30% of the ischio femoral length: 100BP and 30BP). Respiratory disturbance to posture was varied using two respiratory rate conditions (quiet breathing (QB), which is the spontaneous rate, and fast breathing (FB) at a rate imposed by a metronome). The results demonstrated that an increased push effort was associated to a higher iEMG level, and induced greater mean deviation (X (p)) and sway path (SP) of antero-posterior CP displacements. It was concluded that postural muscle stiffness reduces postural steadiness. It was suggested that it could be related to a weaker compensation of respiratory disturbance to body posture. PMID- 16697202 TI - Correlation of blood-brain penetration using structural descriptors. AB - Experimental blood-brain partition coefficients (logBB) for a diverse set of 113 drug molecules are correlated with computed structural descriptors using CODESSA PRO and ISIDA programs to give statistically significant QSAR models based respectively, on molecular and on fragment descriptors. The linear correlation CODESSA-PRO five-descriptor model has correlation coefficient R2=0.781 and standard deviation s2=0.123. The 'consensus model' of ISIDA gave R2=0.872 and s2=0.047. The developed models were successfully validated using the central nervous system activity data of an external test set of 40 drug molecules. PMID- 16697203 TI - Synthesis and antibacterial activity of novel C12 ethyl ketolides. AB - A novel series of C(12) ethyl erythromycin derivatives have been discovered which exhibit in vitro and in vivo potency against key respiratory pathogens, including those resistant to erythromycin. The C(12) modification involves replacing the natural C(12) methyl group in the erythromycin core with an ethyl group via chemical synthesis. From the C(12) ethyl macrolide core, a series of C(12) ethyl ketolides were prepared and tested for antibacterial activity against a panel of relevant clinical isolates. Several compounds were found to be potent against macrolide-sensitive and -resistant bacteria, whether resistance was due to ribosome methylation (erm) or efflux (mef). In particular, the C(12) ethyl ketolides 4k,4s,4q,4m, and 4t showed a similar antimicrobial spectrum and comparable activity to the commercial ketolide telithromycin. The in vivo efficacy of several C(12) ethyl ketolides was demonstrated in a mouse infection model with Streptococcus pneumoniae as pathogen. PMID- 16697204 TI - Synthesis and metabolism of naphthyl substituted phosphoramidate derivatives of stavudine. AB - The synthesis of naphthylphosphoramidate derivatives of stavudine was achieved using a four-step procedure. The derivatives were subjected to several different enzymes including lipase, esterase, Subtilisin Carlsberg, and Carica papaya, and their hydrolysis rates were determined. Based on the rates of hydrolysis, we were able to differentiate between the chiralities at the phosphorus center of the phosphoramidate compounds. In addition, lipase was found to distinguish between both alpha and beta forms of the compounds. The superior chiral selectivity shown by lipase toward the naphthyl substituted phosphoramidate derivatives is attributed to the restrictive binding pocket of the lipase. PMID- 16697205 TI - Synthesis of Sansalvamide A derivatives and their cytotoxicity in the MSS colon cancer cell line HT-29. AB - We report the synthesis of thirty-six Sansalvamide A derivatives, and their biological activity against colon cancer HT-29 cell line, a microsatellite stable (MSS) colon cancer cell-line. The thirty-six compounds can be divided into three subsets, where the first subset of compounds contains L-amino acids, the second subset contains D-amino acids, and the third subset contains both D- and L-amino acids. Five compounds exhibited excellent inhibitory activity (>75% inhibition). The structure-activity relationship (SAR) of the compounds established that a single D-amino acid in position 2 or 3 gave up to a 10-fold improved cytotoxicity over Sansalvamide A peptide. This work highlights the importance of residues 2 and 3 and the role of D-amino acids in the extraordinary SAR for this compound class. PMID- 16697206 TI - Synthesis and evaluation of 5,5-diphenylimidazolones as potent human neuropeptide Y5 receptor antagonists. AB - A series of novel 5,5-diphenylimidazolones was synthesized and evaluated for activity against the human neuropeptide Y5 receptor. The 3-pyridyl analog 46 demonstrated an IC(50) of 8.3 nM with a favorable pharmacokinetic profile in rats, but was ineffective in reducing food intake. PMID- 16697207 TI - Artepillin C isoprenomics: design and synthesis of artepillin C isoprene analogues as lipid peroxidation inhibitor having low mitochondrial toxicity. AB - We designed and synthesized isoprene analogues of artepillin C, a major component of Brazilian propolis, and investigated the inhibitory activity on lipid peroxidation of rat liver mitochondria (RLM) and RLM toxicity based on isoprenomics. We succeeded in the synthesis of artepillin C isoprene analogues using regioselective prenylation within the range from 22% to 53% total yield. Reactivity of artepillin C and its isoprene analogues with ABTS (2,2'-Azinobis(3 ethylbenzothiazoline-6-sulfonate)) radical cations showed only a slight difference among the molecules. The isoprene side-chain elongation analogues of artepillin C showed almost the same inhibitory activity against RLM lipid peroxidation as artepillin C. Artepillin C and its isoprene analogues had very weak RLM uncoupling activity. Moreover, artepillin C and its isoprene analogues exhibited a lower inhibitory activity against adenosine 5'-triphosphate (ATP) synthesis by about two orders of magnitude than the effective inhibitory activity against RLM lipid peroxidation. From these results we conclude that artepillin C isoprene analogues could be potent lipid peroxidation inhibitors having low mitochondrial toxicity. We also conclude that elongation of the isoprene side chain of artepillin C to increase lipophilicity had little influence on the inhibitory activity toward RLM lipid peroxidation. PMID- 16697208 TI - Phosphatidylinositol mannosides: synthesis and suppression of allergic airway disease. AB - Phosphatidylinositol mannoside (PIM) extracts from mycobacteria have been shown previously to suppress allergic airway inflammation in mice. To help determine the structural requirements for activity, PIM1(2) (1), PIM1(6) (2) and PIM2 (3) were synthesized and tested for their ability to suppress cellular inflammation in a mouse model of allergic asthma. The synthetic PIMs were all effective in suppressing airway eosinophilia in the asthma model, with PIM1(6) being the most effective. Suppression of all inflammatory cells monitored was observed, indicating a general blockade of cellular inflammation. Non-mannosylated phosphatidylinositol (PI) had no suppressive effect, indicating that at least one alpha-d-mannopyranosyl residue is necessary for activity. The suppressive effect of the three PIM compounds indicates that other members of this set may be of value in treatment of a range of diseases driven by infiltration of inflammatory cells. PMID- 16697209 TI - Topical anti-inflammatory activity of 2alpha-hydroxy pentacyclic triterpene acids from the leaves of Ugni molinae. AB - Leaf extracts of Ugni molinae Turcz. are used in the Chilean cosmetic industry on the assumption that they have decongestant, regenerative, and anti-aging properties. A bioassay-guided fractionation of this plant material showed that some extracts have potent anti-inflammatory activities. Further fractionation led to the isolation and identification of betulinic acid, a mixture of ursolic and oleanolic acids, and the 2alpha-hydroxy derivatives alphitolic, asiatic, and corosolic acids. The latter three were evaluated in vivo in the mouse ear assay for their topical anti-inflammatory activity, inducing inflammation with either arachidonic acid (AA) or 12-O-tetradecanoylphorbol-13 acetate (TPA). Only corosolic acid was active in the AA assay, with similar potency to nimesulide, but all three triterpene acids inhibited TPA-induced inflammation with potencies comparable to that of indomethacin. PMID- 16697210 TI - Heterogeneous expression of hydrocephalic phenotype in the hyh mice carrying a point mutation in alpha-SNAP. AB - The hyh mouse carrying a point mutation in the gene encoding for soluble N ethylmaleimide-sensitive factor (NSF) attachment protein alpha (alpha-SNAP) develops inherited hydrocephalus. The investigation was designed to study: (i) the clinical evolution of hyh mice; (ii) factors other than the alpha-SNAP mutation that may influence the expression of hydrocephalus; (iii) the neuropathological features underlying the different forms of clinical evolution. The study included 3017 mice, 22.4% of which were hydrocephalic. The neuropathological study was performed in 112 mice by use of light and electron microscopy. It was found that maternal- and sex-related factors are involved in the heterogeneous expression of hyh phenotype. The clinical evolution recorded throughout a 4-year period also revealed a heterogeneous expression of the hydrocephalic phenotype. Two subpopulations were distinguished: (i) 70% of mice underwent a rapidly progressive hydrocephalus and died during the first 2 months of life; they presented macrocephaly, extremely large expansion of the ventricles, equilibrium impairment and decreased motor activity. (ii) Mice with slowly progressive hydrocephalus (30%) survived for periods ranging between 2 months and 2 years. They had no or moderate macrocephaly; moderate ventricular dilatation and preserved general motor activity; they all presented spontaneous ventriculostomies communicating the ventricles with the subarachnoid space, indicating that such communications play a key role in the long survival of these mice. The hyh mutant represents an ideal animal model to investigate how do the brain "adapt" to a virtually life-lasting hydrocephalus. PMID- 16697211 TI - Granulocyte growth factors in the treatment of non-small cell lung cancer (NSCLC). AB - Neutropenia and subsequent infections are common events that limit treatment of non-small cell lung cancer (NSCLC). Granulocyte growth factors (G- and GM-CSF) have been introduced in clinical practice and their use has yielded a reduction of the infection risk related to chemotherapy and a dose increase of drug delivery. Randomized clinical trials have shown that granulocyte colony stimulating factors and, more recently, the longer-acting pegylated granulocyte colony-stimulating factor (pegfilgrastim) effectively reduce the incidence and severity of neutropenia and of its complications. Recommendations for the use of haematopoietic colony-stimulating factors from the American Society of Clinical Oncology (ASCO) have been published in 1994 and updated in 1996, 1997 and 2000. Recently, moreover, National Comprehensive Cancer Network (NCCN) guidelines for the myeloid growth factors in cancer treatment make available. Chemotherapy associated myelosuppression is a major limitation of anticancer therapy also in early stage, local advanced and metastatic NSCLC. Recently, dose-dense chemotherapy has been shown to improve the outcome in early stage breast cancer and non-Hodgkin's lymphoma. However, few randomized trials have been reported on chemotherapy with or without granulocyte growth factors as primary prophylaxis in NSCLC. Presently, there is no evidence for a benefit in response rate and survival from the use of granulocyte growth factors as support of chemotherapy, in particular, for locally advanced and metastatic NSCLC. In clinical practice, the role of granulocyte growth factors for NSCLC treatment should be limited following the guidelines. An appropriate use of granulocyte growth factors may reduce the overall cost of treatment and improve the quality of life, important aims in the treatment of patients with local advanced or metastatic NSCLC. In the future, we need to identify patients who can benefit from granulocyte growth factors for optimize the schedule and doses, in advanced disease and also, after the recent positive results of adjuvant chemotherapy, in early stages. This review summarizes the present knowledge on the use of granulocyte growth factors in NSCLC. PMID- 16697212 TI - Increase of CXC chemokine CXCL10 and CC chemokine CCL2 serum levels in normal ageing. AB - No study has evaluated contemporaneously serum CXC and CC chemokines changes in normal ageing. Serum levels of CXCL10 (sCXCL10) (CXC) and CCL2 (sCCL2) (CC) prototype chemokines have been measured in 164 healthy subjects, from 10 to 79 years of age (82 males/82 females). By simple regression analysis, sCXCL10 and sCCL2 were significantly related with increasing age (r=0.32, p<0.001; r=0.31, p<0.0001, respectively), and with each other (r=0.30, p=0.0004). In a multiple linear regression model, only age and sCCL2 were significantly related to sCXCL10 levels (p<0.001); age and sCXCL10 were significantly related to sCCL2 levels (p<0.001). Subjects with high sCXCL10 levels (>150 pg/ml) were not significantly associated with those with high sCCL2 levels (>559 pg/ml). This study, performed in healthy subjects on an age gradient, demonstrates an increase of sCXCL10 and sCCL2 with advancing age; the differential increase of sCXCL10 or sCCL2 may reflect a general shift towards Th1 or Th2 cytokines pattern, respectively. PMID- 16697213 TI - HMGB1 activates replication of latent HIV-1 in a monocytic cell-line, but inhibits HIV-1 replication in primary macrophages. AB - High mobility group box protein 1 (HMGB1) is an abundant component of mammalian cells that can be released into extracellular milieu actively or by cells that undergo necrosis. Exposure of inflammatory and endothelial cells to HMGB1 leads to the release of cytokines, including TNF-alpha and IL-6. To evaluate the impact of exogenous HMGB1 on viral replication in HIV-1 infected cells, we studied models of latent and acute infection. Extracellular HMGB1 dose dependently increased HIV-1 replication in the monocytic cells, U1, which is an established model for studying latent HIV-1 infection. Dexamethasone, a known inhibitor of NF kappaB signaling in U1 cells, inhibited HMGB1-induced stimulation of the viral production. Addition of HMGB1 to primary monocytic cells with active HIV-1 infection elicited the opposite effect, due to suppression of the viral replication. The mechanism of this unexpected finding was explained by an HMGB1 mediated increased release of chemokines (RANTES, MIP-1alpha, and MIP-1beta) that are known to inhibit HIV-1 replication. The stimulatory effect of the HMGB1 was not present when latently infected T-cells (ACH-2) were used as target cells. Our data suggest that extracellular HMGB1 has a dichotomic effect on the HIV-1 infection in monocytes but not in lymphocytes. Both activation of latent HIV-1 infection and inhibition of active replication can thus be seen in vitro. PMID- 16697214 TI - 14-3-3 proteins and cancer biology. PMID- 16697215 TI - Regulation of the p53-MDM2 pathway by 14-3-3 sigma and other proteins. AB - The 14-3-3 sigma (sigma) protein, a unique member of 14-3-3 family, is a negative regulator of the cell cycle and is induced by p53 to initiate cell cycle checkpoint control after DNA damage. Among the 14-3-3 family members, 14-3-3 sigma is uniquely induced by p53 and has a positive feedback effect on p53 activity in response to DNA damage. Although 14-3-3 sigma is linked to p53 regulated cell cycle checkpoint control, the detailed mechanisms of cell cycle regulation by 14-3-3 sigma remain unclear. Decreased expression of 14-3-3 sigma was reported in several types of carcinomas, suggesting that the negative regulatory role of 14-3-3 sigma in the cell cycle is compromised during tumorigenesis. Given the fact that p53's tumor suppressive function is lost in almost half of all human cancers and that 14-3-3 sigma's activity is linked to the p53 network, a perspective regarding the p53/14-3-3 sigma relationship is needed for cancer research. Here we discuss the mechanisms by which 14-3-3 sigma stabilizes p53 with the hope that these insights may be applied to develop targeted therapeutic strategies for cancer treatment. PMID- 16697216 TI - Dynamic 14-3-3/client protein interactions integrate survival and apoptotic pathways. AB - The serine/threonine binding protein, 14-3-3, possesses a diverse array of client proteins. It is involved in the regulation of apoptosis through multiple interactions with proteins of the core mitochondrial machinery, pro-apoptotic transcription factors, and their upstream signaling pathways. 14-3-3 coordinates with survival kinases to inhibit multiple pro-apoptotic molecules. One prominent mechanism for the suppression of apoptosis is through 14-3-3-mediated sequestration of pro-apoptotic client proteins. On the other hand, cellular stresses appear to signal through the inhibition of 14-3-3 function to exert their pro-apoptotic effect. Global inhibition of 14-3-3/client protein interaction induces apoptosis, and stands as an attractive intervention in diseases involving overactive survival signaling pathways. Because dysregulation of 14-3-3 has been associated with poor survival of cancer patients, targeting 14 3-3 may provide a novel therapeutic approach for the treatment of cancer. PMID- 16697217 TI - Intrathecally infused antibodies against Nogo-A penetrate the CNS and downregulate the endogenous neurite growth inhibitor Nogo-A. AB - Neutralizing antibodies against the neurite growth inhibitory protein Nogo-A are known to induce regeneration, enhance compensatory growth, and enhance functional recovery. In intact adult rats and monkeys or spinal cord injured adult rats, antibodies reached the entire spinal cord and brain through the CSF circulation from intraventricular or intrathecal infusion sites. In the tissue, anti-Nogo antibodies were found inside Nogo-A expressing oligodendrocytes and neurons. Intracellularly, anti-Nogo-A antibodies were colocalized with endogenous Nogo-A in large organels, some of which containing the lysosomal marker cathepsin-D. This suggests antibody-induced internalization of cell surface Nogo-A. Total Nogo A tissue levels in spinal cord were decreased in intact adult rats following 7 days of antibody infusion. This mechanism was confirmed in vitro; cultured oligodendrocytes and neurons had lower Nogo-A contents in the presence of anti Nogo-A antibodies. These results demonstrate that antibodies against a CNS cell surface protein reach their antigen through the CSF and can induce its downregulation. PMID- 16697218 TI - Pin1 allows for differential Tau dephosphorylation in neuronal cells. AB - Neurofibrillary degeneration is likely to be related to abnormal Tau phosphorylation and aggregation. Among abnormal Tau phosphorylation sites, pThr231 is of particular interest since it is associated with early stages of Alzheimer's disease and is a binding site of Pin1, a peptidyl-prolyl cis/trans isomerase mainly involved in cell cycle regulation. In the present work, Pin1 level was found strongly increased during neuronal differentiation and tightly correlated with Tau dephosphorylation at Thr231. Likewise, we showed in cellular model that Pin1 allowed for specific Tau dephosphorylation at Thr231, whereas other phosphorylation sites were unchanged. Moreover, cells displaying Tau phosphorylation at Thr231 did not show any Pin1 nuclear depletion. Altogether, these data indicate that Pin1 has key function(s) in neuron and is at least involved in the regulation of Tau phosphorylation at relevant sites. Hence, Pin1 dysfunction, unlikely by nuclear depletion, may have critical consequences on Tau pathological aggregation and neuronal death. PMID- 16697219 TI - The nuclear lamina and its proposed roles in tumorigenesis: projection on the hematologic malignancies and future targeted therapy. AB - The nuclear lamina, a network of lamin filaments and lamin-associated proteins, is located between the inner nuclear membrane and the peripheral chromatin. The nuclear lamina is involved in numerous nuclear functions including maintaining nuclear shape, determining nuclear positioning, organizing chromatin and regulating the cell cycle, DNA replication, transcription, cell differentiation, apoptosis, and aging. Alterations in the composition of nuclear lamins and their associated proteins are currently emerging as an additional event involved in malignant transformation, tumor propagation and progression, thus identifying potential novel targets for future anti-cancer therapy. Here, we review the current knowledge on lamin expression patterns in cells of hematologic malignancies and give an overview on the roles of the nuclear lamina proteins in heterochromatin organization, apoptosis, and aging with special emphasis on the relevance in cancer development. PMID- 16697220 TI - Shape and compaction of Escherichia coli nucleoids. AB - The genomic DNA in cells of Escherichia coli is localized in one or two compact, phase-like regions with characteristic shapes. Nucleoids undergo progressive changes in shape and compaction in the presence of drugs such as chloramphenicol or puromycin. Forces which influence nucleoid shape and compaction are reviewed, with particular emphasis on crowding effects of the cytoplasm and confinement effects of the cell envelope. Based in part on the theory of Kornyshev and Leikin for interaction between DNA duplexes, the folding of DNA caused by binding of DNA associated proteins is suggested to antagonize DNA condensation and, thereby, increase access to DNA sequences. These views are incorporated into a working model for nucleoid organization. PMID- 16697221 TI - Template-based coiled-coil antigens elicit neutralizing antibodies to the SARS coronavirus. AB - The Spike (S) glycoprotein of coronaviruses (CoV) mediates viral entry into host cells. It contains two hydrophobic heptad repeat (HR) regions, denoted HRN and HRC, which oligomerize the S glycoprotein into a trimer in the native state and when activated collapse into a six-helix bundle structure driving fusion of the host and viral membranes. Previous studies have shown that peptides of the HR regions can inhibit viral infectivity. These studies imply that the HR regions are accessible and that agents which can interact with them may prevent viral entry. In the present study, we have investigated an approach to generate antibodies that specifically recognize the HRN and HRC regions of the SARS-CoV spike (S) glycoprotein in order to evaluate whether these antibodies can inhibit viral infectivity and thus neutralize the SARS-CoV. In this regard, we incorporated HRN and HRC coiled-coil surface residues into a de novo designed two stranded alpha-helical coiled-coil template for generating conformation-specific antibodies that recognize alpha-helices in proteins (Lu, S.M., Hodges, R.S., 2002. J. Biol. Chem. 277, 23515-23524). Eighteen surface residues from two regions of HRN and HRC were incorporated into the template and used to generate four anti-sera, HRN1, HRN2, HRC1, and HRC2. Our results show that all of the elicited anti-sera can specifically recognize HRN or HRC peptides and the native SARS-CoV S protein in an ELISA format. Flow cytometry (FACS) analysis, however, showed only HRC1 and HRC2 anti-sera could bind to native S protein expressed on the cell surface of Chinese hamster ovary cells, i.e., the cell surface structure of the S glycoprotein precluded the ability of the HRN1 or HRN2 anti-sera to see their respective epitope sites. In in vitro viral infectivity assays, no inhibition was observed for either HRN1 or HRN2 anti-serum, whereas both HRC1 and HRC2 anti-sera could inhibit SARS-CoV infection in a dose-dependent manner. Interestingly, the HRC1 anti-serum, which was a more effective inhibitor of viral infectivity compared to HRC2 anti-serum, could only bind the pre-fusogenic state of HRC, i.e., the HRC1 anti-serum did not recognize the six-helix bundle conformation (fusion state) whereas HRC2 anti-serum did. These results suggest that antibodies that are more specific for the pre-fusogenic state of HRC may be better neutralizing antibodies. Overall, these results clearly demonstrate that the two-stranded coiled-coil template acts as an excellent presentation system for eliciting helix-specific antibodies against highly conserved viral antigens and HRC1 and HRC2 peptides may represent potential candidates for use in a peptide vaccine against the SARS-CoV. PMID- 16697222 TI - Tissue specific differences in fibrillin microfibrils analysed using single particle image analysis. AB - Fibrillin microfibrils endow mammalian connective tissues with elasticity and play a fundamental role in the deposition of elastin. The microfibrils are 57 nm periodic supramolecular protein polymers with a mass of 2.5 MDa per repeat. The organisation of molecules within a microfibril is still open to debate and structural studies are only just starting to unravel this issue. The contribution of microfibril associated proteins to microfibril ultrastructure and whether there are any tissue specific differences in microfibril structure is still unknown. Therefore, we have used low dose electron microscopy, single particle image analysis and atomic force microscopy to study the structure of fibrillin microfibrils from different tissues. EM images of microfibrils from aorta, ciliary zonules and vitreous humor were collected and more than 500 microfibril repeats from each sample were subjected to averaging. Averages from each sample were analysed using axial stain exclusion patterns and difference images to detect any variations between them. The overall morphology of fibrillin microfibrils was conserved between tissues and there were only very minor differences in the bead and shoulder region of microfibrils. These data suggest that the structure of isolated microfibrils represents the fibrillin scaffold, and either microfibril associated molecules are lost on purification or play only a minor role in microfibril structure. PMID- 16697223 TI - Synthesizing average 3D anatomical shapes. AB - Average shape estimates are often used to characterize normal morphological variation and discriminate dysmorphology in populations. The purpose of this paper is to estimate "average" or the most representative shapes in populations by using high-resolution medical images as input. The "average" shape is computed from high-dimensional spatial transformations used to co-register each subject in the population rather than the image intensities. Inverse consistent image registration is used to help minimize correspondence errors and produce better population average estimates. Testing the method was done using a population of adult MR brain scans from 22 individuals with no know structural abnormalities. Population averages were computed using the spatial transformation method and local changes in morphology were mapped. Results suggest that this method is a feasible means for robust estimation of population average shape. It is also shown that using inverse consistent transformations produces average shape estimates with less error compared to those produced with transformations with nontrivial inverse consistent errors. PMID- 16697224 TI - Reproducibility of activation in Broca's area during covert generation of single words at high field: a single trial FMRI study at 4 T. AB - Although functional magnetic resonance imaging (FMRI) has arguably become the most ubiquitously used imaging modality, questions remain about the reproducibility of the observed patterns of activation and the acquisition time required to achieve statistically significant and reproducible maps. In the current study, we investigated the reliability of activation in Broca's area, on both a voxel-wise and region of interest level, in response to the covert generation of a single word at 4 T. We also assessed the effects of different parametric (P < 0.01; P < 0.005; P < 0.001) and spatial thresholds (25%, 50% and 75%) on the reproducibility of activation within our region of interest and other randomly selected areas of the brain. We report that the inter-trial consistency of activation within Broca's area for a single trial design using multi-echo EPI is roughly equivalent to previous studies that averaged across a much larger number of trials. However, reliability estimates varied dramatically (approximately 55%) depending on the different parametric and spatial criteria thresholds that were applied to the data. These results show that increased sensitivity at high field strength can be used to reduce the time needed to localize functional activation patterns, which is beneficial for clinical studies such as pre-surgical mapping. Additional benefits of single trial designs, such as the ability to immediately assess for extraneous cognitive processes, are also discussed. PMID- 16697225 TI - Hypothenar Hammer Syndrome: rare or underdiagnosed? AB - Hypothenar hammer Syndrome (HHS) is a condition characterised by digital ischaemia as a result of repetitive trauma to the hypothenar eminence of the hand. It occurs in people who repeatedly use the palm of the hand as a hammer to push, grind or twist objects. It is a curable and a preventable cause of upper digital ischemia. In this report we present a case of HHS and discuss the causes and pathogenesis of this syndrome. We review the incidence, clinical characteristics, differential diagnosis, investigation and treatment. PMID- 16697226 TI - Indirect detection of nuclear magnetic resonance via geometrically induced long range dipolar fields. AB - We report the indirect detection of the magnetization of one spin species via the NMR signal of a second species. Our method relies on the control of long-range dipolar fields between two separate objects, in this case, a water droplet (sensor) immersed in a tube containing mineral oil (sample). Unlike prior experiments, no gradient pulses are used; rather, the setup geometry is exploited to select the part of the sample to be probed and modulate the spin alignment in the sensor. Our results are discussed in the context of Dipolar Field Microscopy, a proposed strategy in which the detector is a hyperpolarized tip. PMID- 16697227 TI - Homozygous nonsense mutation in the MCEE gene and siRNA suppression of methylmalonyl-CoA epimerase expression: a novel cause of mild methylmalonic aciduria. AB - Methylmalonyl-CoA epimerase (MCE) catalyzes the interconversion of D- and L methylmalonyl-CoA in the pathway responsible for the degradation of branched chain amino acids, odd chain-length fatty acids, and other metabolites. Despite the occurrence of metabolic disorders in the enzymatic step occurring immediately upstream of MCE (propionyl-CoA carboxylase) and downstream of MCE (adenosylcobalamin-dependent methylmalonyl-CoA mutase), no disease-causing mutations have been described affecting MCE itself. A patient, formerly identified as belonging to the cblA complementation group of vitamin B12 disorders but lacking mutations in the affected gene, MMAA, was tested for mutations in the MCEE gene. The patient's fibroblasts had normal levels of adenosylcobalamin compared to controls, whereas other cblA cell lines typically had reduced levels of the cofactor. As well, this patient had a milder form of methylmalonic aciduria than usually observed in cblA patients. The patient was found to be homozygous for a c.139C>T (p.R47X) mutation in MCEE by sequence analysis that was confirmed by restriction digestion of PCR products. One sibling, also with mild methylmalonic aciduria, was homozygous for the mutation. Both parents and one other sibling were heterozygous. A nearby insertion polymorphism, c.41-160_161insT, heterozygous in both parents, showed the wild type configuration on the mutant alleles. To assess the impact of isolated MCE deficiency in cultured cells, HeLa cells were transfected with a selectable vector containing MCEE-specific small interfering RNA (siRNA) to suppress gene expression. The reduced level of MCEE mRNA resulted in the reduction of [14C] propionate incorporation into cellular macromolecules. However, siRNA only led to a small reduction in pathway activity, suggesting that previously postulated non enzymatic conversion of D- to L-methylmalonyl-CoA may contribute to some flux through the pathway. We conclude that the patient's MCEE defect was responsible for the mild methylmalonic aciduria, confirming a partial requirement for the enzymatic activity in humans. PMID- 16697228 TI - The impact of comparative genomics on infectious disease research. AB - The past decade has witnessed a revolution in infectious disease research, fuelled by the accumulation of a huge amount of DNA sequence data. The avalanche of genome sequence information has largely promoted the development of comparative genomics, which exploits available genome sequences to perform either inter- or intra-species comparisons of bacterial genome contents, or performs comparisons between the human genome and those of other organisms. This review aims to summarize how comparative genomics is being extensively used in infectious disease research, such as in the studies to identify virulence determinants, antimicrobial drug targets, vaccine candidates and new markers for diagnostics. These applications hold considerable promise for alleviating the burden of infectious diseases in the coming years. PMID- 16697229 TI - The impact of genomics on the analysis of host resistance to infectious disease. AB - The advent of new technologies and resources, including the complete sequence of mammalian genomes, has had a dramatic impact on the genetic analysis of susceptibility to infections in humans and in animal models of infection. Genes responsible for simple or complex control of susceptibility to infection with different pathogens have been recently identified and characterized, and are reviewed herein. PMID- 16697230 TI - Matrix metalloproteinase-9 (gelatinase B) deficiency leads to increased severity of Staphylococcus aureus-triggered septic arthritis. AB - Matrix metalloproteinases constitute a family of structurally related endopeptidases that are crucial for the normal turnover of the extracellular matrix. Elevated levels of MMP-9 have been demonstrated in synovial fluids of rheumatoid arthritis patients, and a correlation with the severity of the disease has been described. The aim of this study was to explore the impact of MMP-9 expression on joint inflammation and destruction in a model of bacterially induced septic arthritis. MMP-9 knock-out mice and C57Bl6 congenic controls were inoculated intravenously or intra-articularly with Staphylococcus aureus. Arthritis was evaluated clinically and by means of histology. Zymographic analyses were performed to study ex vivo induction of MMP-9 following exposure to S. aureus. The MMP-9 knock-out mice displayed a significantly higher frequency and severity, but not destructivity, of arthritis than did the wild-type mice. The knock-out mice also proved to harbour an increased number of bacteria locally in joints and systemically in kidneys, possibly by impaired extravasation and recruitment of leukocytes and a deficient early defence against infection. Our findings indicate that deficiency in MMP-9 increases the degree of joint inflammation due to decreased bacterial clearance. PMID- 16697231 TI - Augmentation of antimicrobial metronidazole therapy of bacterial vaginosis with oral probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14: randomized, double-blind, placebo controlled trial. AB - This study enrolled 125 premenopausal women diagnosed with bacterial vaginosis (BV) by presence of vaginal irritation, discharge and 'fishy' odor, and Nugent criteria and detection of sialidase enzyme. The subjects were treated with oral metronidazole (500 mg) twice daily from days 1 to 7, and randomized to receive oral Lactobacillus rhamnosus GR-1 (1 x 10(9)) and Lactobacillus reuteri RC-14 (1 x 10(9)) or placebo twice daily from days 1 to 30. Primary outcome was cure of BV as determined by normal Nugent score, negative sialidase test and no symptoms or signs of BV at day 30. A total of 106 subjects returned for 30-day follow-up, of which 88% were cured in the antibiotic/probiotic group compared to 40% in the antibiotic/placebo group (p<0.001). Of the remaining subjects, 30% subjects in the placebo group and none in the probiotic group had BV, while 30% in the placebo and 12% in the probiotic group fell into the intermediate category based upon Nugent score, sialidase result and clinical findings. High counts of Lactobacillus sp. (>10(5) CFU/ml) were recovered from the vagina of 96% probiotic treated subjects compared to 53% controls at day 30. In summary, this study showed efficacious use of lactobacilli and antibiotic in the eradication of BV in black African women. PMID- 16697232 TI - A role for dendritic cells in the dissemination of mycobacterial infection. AB - The ability of mycobacteria to disseminate from the initial site of infection has an important role in immune priming and in the seeding of disease in multiple organs. To study this phenomenon, we used flow cytometry to analyse the distribution of green fluorescent protein-labelled BCG amongst different populations of antigen-presenting cells in the lungs of mice following intranasal infection, and monitored appearance of live bacteria in the draining mediastinal lymph nodes. BCG predominantly infected alveolar macrophages (CD11c(+)/CD11b(-)) and dendritic cells (CD11c(+)/CD11b(+)) in the lungs. The bacteria that disseminated to the lymph node were found in dendritic cells. The results are consistent with a model in which mycobacterial dissemination from the lung is initiated by the migration of infected dendritic cells to the draining lymph nodes. PMID- 16697233 TI - The immunobiology of Acanthamoeba keratitis. AB - Acanthamoeba spp. are free-living amoebae that cause Acanthamoeba keratitis, a blinding corneal infection. The innate immune apparatus is crucial for the resolution of the disease. With the exception of mucosal antibody, elements of the adaptive immune system fail to prevent infection or contribute to its resolution in experimental animals. PMID- 16697234 TI - Adhesion and cytotoxicity of Bacillus cereus and Bacillus thuringiensis to epithelial cells are FlhA and PlcR dependent, respectively. AB - Some bacteria of the Bacillus cereus group are enteropathogens. The first cells encountered by bacteria following oral contamination of the host are epithelial cells. We studied the capacity of these bacteria to adhere to epithelial cells and the consequences of this interaction. We found that cell adhesion is strain dependent and that a strain mutated in flhA, which encodes a component of flagellum-apparatus formation, is impaired in adhesion, suggesting that flagella are important virulence factors. The bacteria are cytotoxic to epithelial cells and induce substantial cytoplasmic and membrane alterations. However, direct contact between cells and bacteria is not required for cytotoxicity. The determinants of this cytotoxicity are secreted and their expression depends on the pleiotropic regulator PlcR. Adhesion and cytotoxicity of B. cereus to epithelial cells might explain the diarrhea caused by these pathogens. Our findings provide further insight into the pathogenicity of B. cereus group members. PMID- 16697235 TI - Parachlamydia acanthamoebae enters and multiplies within pneumocytes and lung fibroblasts. AB - Parachlamydia acanthamoebae is a Chlamydia-like organism that naturally infects free-living amoebae. P. acanthamoebae is a putative emerging agent of community acquired and inhalation pneumonia that may enter and multiply within human macrophages. However, since Parachlamydia induces their apoptosis, macrophages may not represent a perennial niche for this obligate intracellular bacterium. Therefore, we investigated whether pneumocytes and lung fibroblasts are permissive to Parachlamydia infection and might act as a replicative niche. Entry of Parachlamydia into pneumocytes (A549) and lung fibroblasts (HEL) was confirmed by confocal and electron microscopy. In A549 cells, the mean number of Parachlamydia per cell increased 7-fold from day 0 to day 7, independently of the technique used to label the bacteria. The proportion of infected A549 cells also increased over time, whereas cell viability remained unaffected by Parachlamydia infection. The sustained (3 weeks) viability of Parachlamydia when incubated in the presence of A549 cells contrasted with that observed in the absence of cells. HEL cells were also permissive to Parachlamydia infection, as we observed a 3- to 4-fold increase in the mean number of bacteria per cell. In HEL cells, Parachlamydia retained some viability for 2 weeks. These findings demonstrate that Parachlamydia is able to enter and multiply within pneumocytes and fibroblasts. The viability of both cell types was not compromised after Parachlamydia infection. We therefore conclude that these cells may remain infected for a prolonged time and may represent an intrapulmonary niche for the strictly intracellular Parachlamydia. This indirectly supports the role of Parachlamydia as an agent of pneumonia. PMID- 16697236 TI - Decrease in Epstein-Barr virus-positive AIDS-related lymphoma in the era of highly active antiretroviral therapy. AB - Recent introduction of highly active antiretroviral therapy (HAART) is reported to have reduced the incidence of lymphoma among HIV-infected individuals. A clinicopathological study was performed on 86 AIDS-related lymphoma patients who were treated in Tokyo area from 1987 to 2005. The incidence of lymphoma detected by autopsy was 27% (53 cases/198 autopsies). Diffuse large B cell lymphoma was the most predominant histological subtype throughout the period (78%). Burkitt's lymphoma (BL) increased from 2% in the pre-HAART era (before end-1997) to 13% in the HAART era, whereas incidence of BL did not vary between HAART users and non users. Epstein-Barr virus (EBV)-positive lymphoma decreased from 88% in the pre HAART era to 58% in the HAART era, but did not differ significantly between HAART users (73%) and non-users (74%). Nodal involvement of lymphoma increased from 14% in the pre-HAART era to 50% in the HAART era; however, central nervous system involvement decreased from 62 to 38%. Kaposi's sarcoma-associated herpesvirus infection was rare (4%) among all cases. These data suggest that HAART might play a partial role in these changes, and the alteration in immunological backgrounds, such as EBV prevalence, is suggested as another leading cause of these changes in Japanese AIDS-related lymphoma. PMID- 16697237 TI - Emergence of distinct genetic variants in the population of primary Bartonella henselae isolates. AB - Bartonella henselae isolates from different hosts display a marked genetic heterogeneity, as determined by pulsed-field gel electrophoresis (PFGE). The aim of the present study was to determine whether different genetic variants may coexist within the population of distinct B. henselae isolates and could be detected by PFGE. Three primary B. henselae isolates and the B. henselae reference strains ATCC 49793 and 49882 were subjected as single colony derived cultures in quadruplicate to PFGE analysis upon restriction with SmaI or NotI. Up to 4 fragment differences were found among the cultures obtained from each primary isolate, indicating the coexistence of genetic variants in the population of primary B. henselae isolates. The clonal relatedness of the genetic variants was confirmed by arbitrarily primed PCR and multi-locus sequence typing. In contrast to the primary isolates, no variants were detected among the single colony derived cultures of the high-passage ATCC strains. We hypothesized that the coexistence of different genetic variants may represent a feature that is restricted to primary or low-passage B. henselae isolates. The primary isolates were serially passed in vitro and then subjected as single colony derived cultures to PFGE analysis, which now revealed identical patterns among the quadruplicate cultures of each high-passage isolate. These results suggest that the population of a primary B. henselae isolate is composed of distinct genetic variants, which may disappear upon repeated passages on artificial culture media. Generation of genetic variants by B. henselae may represent an escape mechanism to circumvent the host specific immune responses. PMID- 16697238 TI - Introduction: genomic approaches in infectious diseases. PMID- 16697239 TI - Dividing oceans into pools: strategies for the global analysis of bacterial genes. AB - In bacterial pathogenesis, it is often easy to accept the results of large-scale screens without independent verification of the results. How can one critically read this literature? Here we review issues inherent in genome-wide screens in bacteria, focusing on experiments that attempt to comprehensively identify genes required for bacterial growth under specific conditions. Our analysis suggests that the methodologies employed undoubtedly shape the results. It is clear, however, that the question is not which method is better but which provides the data most suited to a given question. PMID- 16697240 TI - [SERMs for treatment of osteoporosis: state-of-the-art and perspectives]. AB - Every year in France, we observe 60,000 vertebral fractures, 50,000 hip fractures and 35,000 wrist fractures. However, only 20% of these patients receive adequate pharmacologic therapy to prevent new fractures and their complications. At the present time, clinicians have not a perfect knowledge of the therapeutic class of SERMs (Selective Estrogen Receptor Modulators). In this paper, the authors show the strong capacity of these drugs to produce, in osteoporotic postmenopausal women, major therapeutic effects on bone (reduction of fracture risk), on breast (reduction of cancer risk), and without any side effects on arterial diseases except an increased risk of venous thromboembolism. PMID- 16697241 TI - Ultrasound accelerated esterification of palmitic acid with vitamin C. AB - The esterification of palmitic acid with vitamin C in the presence of concentrated sulfuric acid as the solvent and catalyst by means of 25 kHz ultrasonic irradiation to obtain l-ascorbyl 6-palmitate is studied. By using ultrasound the dissolution rate of the reactants can be accelerated greatly, the reaction time of esterification can be reduced from 36 to 2h, and better yield (90-93%) of ester can be given by using 95% concentrated sulfuric acid as the solvent and catalyst, contrast to the yield of 75-85% by using 99% concentrated sulfuric acid without ultrasound. The influence of reaction conditions and ultrasonic parameters to the yield of ascorbyl palmitate are reported. PMID- 16697242 TI - Langerhans cells: mediators of immunity and tolerance. AB - Langerhans cells provide the epidermis with a surveillance network that samples the external environment influencing the decision between immunity and tolerance. Langerhans cells are immature dendritic cells acquiring antigens from foreign invaders as well as damaged native tissue for display to the immune response. The current paradigm suggests that the state of maturity of Langerhans cells, defined by the display of molecules that provoke immune responses (histocompatibility, co stimulators, adhesion and homing receptors), determines whether emigration of the Langerhans cell to lymph nodes signals immunity or tolerance. Other factors such as type of immunogen ingested, environmental danger signals and the level of cell death may also play a role in tipping the balance towards immunity or immunosuppression. As modulators of the immune response, Langerhans cells play a role in cutaneous autoimmunity in lupus and in cancers that have an affinity for the epidermis such as cutaneous T cell lymphoma. PMID- 16697243 TI - In vitro denaturation-renaturation of fibronectin. Formation of multimers disulfide-linked and shuffling of intramolecular disulfide bonds. AB - It is well established that fibronectin into extracellular matrix undergoes repeated tensions applied by cells, resulting into dramatic structural changes which reflect its elastic properties. However, there is currently no study reporting with precision the consequences of this elasticity on fibronectin structure and conformation. In the present work, we investigated fibronectin structural and conformational reorganization in vitro through a denaturation renaturation approach. The similarities and differences between "refolded fibronectin" and "native fibronectin" were investigated using various spectroscopic methods, hydrodynamic characterization, molecular imaging and biochemical characterization. In the refolded form, secondary structure elements as well as local tyrosine and tryptophan environment are identical compared to the native form. Interestingly, some differences in global tertiary structure organization and molecular conformation were observed. These differences are due to the reactivity of the two free cysteines, which are buried in the native state but become accessible during the unfolding process. First, oxidation of these residues leading to the formation of intermolecular disulfide bonds results in formation of stabilized multimer. Second, some illegitimate intramolecular disulfide bonds are formed. The presence of iodoacetamide, the sulfhydryl alkylating agent, during the unfolding-refolding process prevents all these events. This study clearly demonstrates that, under near physiological conditions, competitive renaturation pathways occur, involving free cysteines in either multimer formation or intermolecular shuffling of disulfide bonds. These findings might have important implications for future studies and be helpful to develop a deeper understanding of fibronectin morphology. PMID- 16697244 TI - Mechanistic models for wind dispersal. AB - The growing need for ecological forecasts of, for example, species migration, has increased interest in developing mechanistic models for wind dispersal of seeds, pollen and spores. Analytical models are only able to predict mean dispersal distances, whereas sophisticated trajectory simulation models are able to incorporate rare wind conditions causing long-distance dispersal and are therefore preferable. Despite the rapid development of mechanistic dispersal models, only a few studies have focused on comparing the performance of the models. To assess the level of model complexity needed, attention should be paid to model comparisons and the sensitivity of the predictions to model complexity. In addition to studying the movement of airborne particles, future modelling work should also focus on the processes of particle release and deposition. PMID- 16697245 TI - Ammonia: a candidate for nitrogen transfer at the mycorrhizal interface. AB - In mycorrhizal associations, the fungal partner assists its plant host with nitrogen and phosphorus uptake while obtaining photosynthetically fixed carbon. Recent studies in mycorrhiza have highlighted the potential for direct transfer of ammonia from fungal to plant cells. This presents a new perspective on nitrogen transfer at the mycorrhizal interface, which is discussed here in light of recent progress made in characterizing a large array of membrane proteins that could fulfil the function of transporting ammonia. PMID- 16697246 TI - Wild sex in the grasses. AB - To date, alien introgression of agronomically important traits into bread wheat (Triticum aestivum) from wild relatives has not been readily achievable through traditional breeding practices. However, this door might now be unlocked. The insightful research published recently by Graham Moore and his team delivers a likely candidate in the form of a cdc2-kinase-related gene family for the Ph1 locus--a chromatin region located on chromosome 5B that is responsible for homologous chromosome pairing integrity in bread wheat. PMID- 16697247 TI - Trichomes: different regulatory networks lead to convergent structures. AB - Sometimes, proteins, biological structures or even organisms have similar functions and appearances but have evolved through widely divergent pathways. There is experimental evidence to suggest that different developmental pathways have converged to produce similar outgrowths of the aerial plant epidermis, referred to as trichomes. The emerging picture suggests that trichomes in Arabidopsis thaliana and, perhaps, in cotton develop through a transcriptional regulatory network that differs from those regulating trichome formation in Antirrhinum and Solanaceous species. Several lines of evidence suggest that the duplication of a gene controlling anthocyanin production and subsequent divergence might be the major force driving trichome formation in Arabidopsis, whereas the multicellular trichomes of Antirrhinum and Solanaceous species appear to have a different regulatory origin. PMID- 16697248 TI - Induction of sister chromatid exchanges in traffic policemen exposed to vehicular exhaust. AB - In urban areas there is an explosive growth of population and the number of automobiles. The ever-increasing vehicular traffic density is posing continued threat to the ambient air quality. Traffic policemen as a group of workers are exposed occupationally to the pollutants from vehicular exhaust. Sister chromatid exchanges (SCEs) as a biomarker of the pollutant's effect, were analyzed in peripheral blood lymphocytes of 85 traffic policemen and 60 control subjects. There was a significant increase in the mean SCEs+/-S.D./cell in the exposed group (9.31+/-5.29) when compared to the controls (4.18+/-1.85). Thus the present study concludes that vehicular exhaust might induce cytogenetic damage in traffic police. Further, the more pronounced frequency of SCEs observed in the smoking traffic policemen than in the non-smoking group suggests the joint effect of smoking and vehicular exhaust. PMID- 16697249 TI - Modification of cyclophosphamide-induced clastogenesis and apoptosis in rats by alpha-lipoic acid. AB - The aim of the present study was to investigate the protective efficacy of alpha lipoic acid (LA) on the cyclophosphamide (CP)-induced chromosomal aberrations (CA) and apoptosis in the bone marrow of rats. Male Wistar rats of 140+/-20 g were categorized into eight groups. Five groups were administered CP (40 mg/kg body weight, intraperitoneally) to induce toxicity; four of these groups received a single intraperitoneal injection of LA at a dose of either 100 or 200 mg/kg body weight, and either 30 or 60 min prior to CP administration. A vehicle treated control group and LA control groups were also included. Twenty-four hours after CP treatment, the frequency of CA in bone marrow cells were significantly increased in comparison with the controls. The CP-induced CA were associated with significant increase in DNA damage in the bone marrow as evidenced by increased single strand breaks, whereas in rats treated with LA and CP, the frequency of CA and single strand breaks were significantly decreased in comparison to those given CP alone. CP administration distinctly triggered the apoptotic and necrotic cell death, and LA pretreatment affected cell death by decreasing the number of apoptotic and necrotic cells. The protective effect of LA was found to be stronger at a dose of 200 mg/kg body weight than 100 mg/kg body weight dosage, indicating the dose dependent protective effect of LA. However, the protection by LA was not dependent on the time intervals between LA and CP administration. The results of this study illustrate the protective effect of LA on the CA and apoptosis induced by CP in the erythropoietic system of rats. PMID- 16697250 TI - DNA damage and repair in haemolymph cells of golden mussel (Limnoperna fortunei) exposed to environmental contaminants. AB - The development of methodologies for biomonitoring freshwater ecosystems is of particular relevance in view of the serious problem of aquatic environmental pollution. The mussel species Limnoperna fortunei (golden mussel) was chosen to be tested as a biomonitor organism based on its population data and distribution. L. fortunei individuals were exposed to UV radiation in vitro, and in vivo to pentachlorophenol (PCP) and copper sulphate (CuSO(4)), with the aim of standardizing comet assay and micronucleus test methodologies and evaluating the potential of this organism as a biomonitor. Haemolymph cells immobilized in agarose on slides exposed to UV radiation showed a dose-response relationship with maximum damage at 4.2 J/m(2). For the chemical tests, individuals were exposed for 2h for the comet assay and 24 and 48 h for the micronucleus test. A dose-response relationship was observed for both chemicals. 3x10(-5) M CuSO(4) induced high genotoxicity, also producing some toxicity after 48 h of exposure. PCP induced maximum damage in both assays at 150 MUg/L. Individuals exposed to PCP showed 100% repair 2 h after the exposure period, as assessed by the comet assay. Exposure to an environmental sample over 7 days confirmed the mussel sensitivity to water contaminants, detected both by the comet assay and the micronucleus test. The results allow us to suggest the golden mussel as a potential biomonitor organism. PMID- 16697251 TI - Polarized IR and Raman spectra and ab initio calculations for bis(guanidine) zirconium bis(nitrilotriacetate) hydrate single crystal [C(NH2)3]2[Zr[N(CH2COO)3]2](H2O)--the new laser Raman converter. AB - Fourier transform polarized IR and Raman spectra of bis(guanidine) zirconium bis(nitrilotriacetate) hydrate single crystal [C(NH(2))(3)](2)[Zr[N(CH(2)COO)(3)](2)](H(2)O) have been measured in the regions 30-4000 and 80-4000 cm(-1) and correlated with X-ray structural data. The factor group analysis has been applied in the discussion of the dichroic dependence of the vibrational modes. The assignment of the internal vibrations for the [Zr(nitrilotriacetate)2]2- complex ion has been based on the ab initio quantum chemical calculations. The usefulness of the studied crystal as Raman laser converter was analyzed basing on the comparison of the spontaneous and stimulated Raman spectra. PMID- 16697252 TI - Diffuse interstellar bands: a comprehensive laboratory study. AB - As a result of the search for the identity of the chromophores responsible for producing the diffuse interstellar bands, a comprehensive exposition of experimental data is presented, which implicates the following molecules: (1) The extremely stable organic molecules, magnesium tetrabenzoporphyrin (MgTBP) and H(2)TBP. (2) A paraffin matrix (referred to as "grains") containing TBPs. (3) A low concentration of pyridine (also within the grains), whose transmission window at 2175 A, accounts for the ubiquitous UV bump. The blue emission spectra associated with the central star, HD44179, of the Red Rectangle displays the fluorescence excitation spectra of bare MgTBP. This unique spectrum matches the low temperature lab data of MgTBP in the vapor phase. An effective grain temperature of 2.728 K (+/-0.008) was deduced, based on MgTBP's lowest measured vibrational state of 341 GHz. PMID- 16697253 TI - Interobserver variation in the interpretation of SSEPs in anoxic-ischaemic coma. AB - OBJECTIVE: To study interobserver variation in the interpretation of median nerve SSEPs in patients with anoxic-ischaemic coma. METHODS: SSEPs of 56 consecutive patients with anoxic-ischaemic coma were interpreted independently by 5 experienced clinical neurophysiologists using guidelines derived from a pilot study. Interobserver agreement was expressed as kappa coefficients. RESULTS: Kappa ranged from 0.20 to 0.65 (mean 0.52, SD 0.14). Disagreement was related with noise level and failure to adhere strictly to the guidelines in 15 cases. The presence or absence of N13 and cortical peaks caused disagreement in 5 cases each. For recordings with a noise level of 0.25 microV or more, mean kappa was 0.34; for recordings with a noise level below 0.25 microV mean kappa was 0.74. CONCLUSIONS: Interobserver agreement for SSEPs in anoxic-ischaemic coma was only moderate. Since the noise level strongly influenced interobserver variation, utmost attention should be given to its reduction. If an artefact level over 0.25 microV remains, absence of N20 cannot be judged with sufficient certainty and the SSEP should be repeated at a later stage. SIGNIFICANCE: Because of its moderate interobserver agreement, great care has to be given to accurate recording and interpretation of SSEPs before using the recordings for non-treatment decisions. PMID- 16697254 TI - Metabolic genotypes as modulators of asbestos-related pleural malignant mesothelioma risk: a comparison of Finnish and Italian populations. AB - The role of CYP1A1, GSTM1, GSTT1, EPHX1, and NAT2 genotypes in susceptibility to malignant mesothelioma (MM) was compared in two case-control studies, previously conducted in two countries where different types of asbestos fibers have been used [Hirvonen et al., 1995. Inherited GSTM1 and NAT2 defects as concurrent risk modifiers in asbestos-related human malignant mesothelioma. Cancer Res. 55, 2981 2983; Hirvonen et al., 1996. Glutathione S-Transferase and N-Acetyltransferase genotypes and asbestos-associated pulmonary disorders. J. Natl. Cancer Inst.88, 1853-1856; Neri et al., 2005. Pleural malignant mesothelioma, genetic susceptibility and asbestos exposure. Mutat. Res. 592, 36-44]. Fifty-seven asbestos-exposed MM patients and 255 controls were recruited in Italy, 48 cases and 121 controls in Finland. In order to make the two studies comparable, they have been updated and new genotyping analyses have been performed. The NAT2 fast acetylator and EPHX1 low-activity genotypes were positively associated with MM in the Italian study, while they were negatively associated with this malignancy in the Finnish one. A combined significant effect was also observed in the Italian study for the NAT2 fast acetylator and EPHX1 low-activity genotypes, while this combination was protective in the Finnish study. Combination of NAT2 fast acetylator and GSTM1 null genotype posed a significantly increased risk of MM in the Italian, but not in the Finnish study. The opposite results obtained in Finland and Italy may be ascribed to random chance, but a role may be hypothesized for the fact that different types of asbestos have been used in the two countries. PMID- 16697255 TI - Nosocomial respiratory syncytial virus infection: impact of prospective surveillance and targeted infection control. AB - BACKGROUND: Nosocomially acquired respiratory syncytial virus (RSV) infections cause serious problems in hospitalized patients. An increased effort should be made to describe the problems connected with such infections in pediatric hospitals, with the aim of reducing the occurrence of nosocomial RSV infections (NI). METHODS: A specialized database was introduced for surveillance and a multifaceted barrier concept based on the CDC recommendations was developed for the control of NI in a university children's hospital in Germany. RESULTS: Between 1999 and 2002 (November 1-April 30), 283 RSV infections (general population) were prospectively documented. Thirty-nine cases (13.8%) were nosocomial infections (NI) with an incidence density (ID) of 0.99/1000 patient days; 48.7% of all NI were found in prematurely born infants. Following the introduction of a surveillance and prevention policy, a 9-fold decrease of the ID (1.67 vs. 0.18/1000 patient-days) was found when comparing the first and the last season. Intensive care treatment was required in 18% of all documented RSV infections, in 48.7% of all NI cases and in 43.5% of all RSV-infected prematurely born infants. Overall RSV-related mortality was 0.71%. CONCLUSIONS: Early diagnosis, a strict cohorting and contact isolation policy, and prospective surveillance contribute to the reduction of nosocomial RSV infection. PMID- 16697256 TI - Specific factors for prenatal lead exposure in the border area of China. AB - The objectives of this study are to examine the prevalence of increased blood lead concentrations in mothers and their umbilical cords, and to identify risk factors for prenatal lead exposure in Kunming city, Yunnan province, China. The study was conducted at two obstetrics departments, and 100 peripartum women were enrolled. The mean blood lead concentrations of the mothers and the umbilical cords were 67.3microg/l and 53.1microg/l, respectively. In multiple linear regression analysis, maternal occupational exposure, maternal consumption of homemade dehydrated vegetables and maternal habitation period in Kunming city were significantly associated with an increase of umbilical cord blood lead concentration. In addition, logistic regression analysis was used to assess the association of umbilical cord blood lead concentrations that possibly have adverse effects on brain development of newborns with each potential risk factor. Maternal frequent use of tableware with color patterns inside was significantly associated with higher cord blood lead concentration in addition to the three items in the multiple linear regression analysis. These points should be considered as specific recommendations for maternal and fetal lead exposure in this city. PMID- 16697257 TI - Ventricular septal defect closure in right coronary cusp prolapse and aortic regurgitation complicating VSD in the outlet septum: which treatment is most appropriate? AB - BACKGROUND: There is currently not a standardized technique for the sizing and shaping of surgical closure of the ventricular septal defect (VSD) patch in patients with right coronary aortic cusp prolapse and aortic regurgitation (AR) complicating VSD in the outlet septum. METHODS: Forty-six VSD patients who had aortic valve prolapse were divided into groups DC (direct closure, n=19), and SPC (small patch closure, n=27). Preoperative and postoperative echocardiography with Doppler color flow interrogation was performed on all patients. RESULTS: In the DC group, among seven patients who had aortic valve prolapse but no AR preoperative, one patient developed AR during postoperative follow-up period. In the remaining 12 patients who had mild AR associated with aortic valve prolapse prior to the procedure, AR was diminished in four and unchanged in six patients. However, AR was aggravated in two patients who required further operations for AV repair or replacement. In the SPC group, among the eight patients who had no preoperative AR, AR progressed in one patient postoperatively. In the remaining 19 patients who had mild AR, AR was diminished in 15 and unchanged in four. The outcome from the operative procedure was significantly better in the SPC group than DC group with mild preoperative AR (chi(2)=7.82; P<0.05). CONCLUSIONS: Small patch closure for this type of VSD is safer and more reliable in improving mild AR than that of direct closure, especially in patients with mild AR. PMID- 16697258 TI - B cell depletion therapy in systemic rheumatic diseases: different strokes for different folks? AB - Autoantibodies have, until recently, been the overriding focus of investigators of autoantibody-associated diseases. Increasing attention is now being paid to B cells, which not only are the producers of autoantibodies but also contribute to autoimmune disease via autoantibody-independent mechanisms. Therapeutic measures that target B cells for depletion are gaining in popularity. In this review, we will focus on two distinct approaches of depleting B cells; one employing a direct-kill approach by engagement of B cell surface CD20 with an anti-CD20 monoclonal antibody (rituximab), and the other employing an indirect starvation approach by neutralization of B lymphocyte stimulator (BLyS), a potent B cell survival factor. Among the systemic immune-based rheumatic disorders, we will focus on rheumatoid arthritis and systemic lupus erythematosus, two disorders for which therapeutic B cell targeting is being intensely investigated. PMID- 16697259 TI - A new generation of retroviral producer cells: predictable and stable virus production by Flp-mediated site-specific integration of retroviral vectors. AB - We developed a new strategy that provides well-defined high-titer producer cells for recombinant retroviruses in a minimum amount of time. The strategy involves the targeted integration of the retroviral vector into a chromosomal locus with favorable properties. For proof of concept we established a novel HEK293-based retroviral producer cell line, called Flp293A, with a single-copy retroviral vector integrated at a selected chromosomal locus. The vector was flanked by noninteracting Flp-recombinase recognition sites and was exchanged for different retroviral vectors via Flp-mediated cassette exchange. All analyzed cell clones showed correct integration and identical titers for each of the vectors, confirming that the expression characteristics from the parental cell were preserved. Titers up to 2.5 x 10(7) infectious particles/10(6) cells were obtained. Also, high-titer producer cells for a therapeutic vector that encodes the 8.9-kb collagen VII cDNA in a marker-free cassette were obtained within 3 weeks without screening. Thus, we provide evidence that the precise integration of viral vectors into a favorable chromosomal locus leads to high and predictable virus production. This method is compatible with other retroviral vectors, including self-inactivating vectors and marker-free vectors. Further, it provides a tool for evaluation of different retroviral vector designs. PMID- 16697260 TI - Transesophageal echocardiography: a screening method for pulmonary vein stenosis after catheter ablation of atrial fibrillation. AB - AIMS: Pulmonary vein (PV) stenosis has been described as a complication after catheter ablation of atrial fibrillation. The aim of the study was to investigate the diagnostic role of transesophageal echocardiography (TEE) in the assessment of PV stenosis. METHODS: Ninety-one patients (71 men, mean age 57+/-16years), initially treated by catheter ablation of atrial fibrillation, underwent re ablation because of arrhythmia recurrences. PV angiograms and TEE were performed before the first and second ablation. PVs were analysed in an intraindividual comparison by measurements of mean and peak flow velocity and of velocity time integrals and diameters. PV angiograms served as standard for assessment of PV stenosis. RESULTS: Sixteen of 91 patients developed PV stenoses as a consequence of the first ablation (13 mild PV stenoses, 4 moderate PV stenoses). All patients with PV stenosis were asymptomatic. In moderate PV stenosis (50-70%) a significant increase of blood flow parameters, reduction of vessel diameter, inhomogeneous blood flow and aliasing were demonstrated by TEE. Using quantitative TEE criteria moderate PV stenosis could be identified with a sensitivity of 84% and specificity of 98%. Detection of mild PV stenosis (30-50%) is challenging (sensitivity of 48% and specificity of 75%). CONCLUSIONS: TEE identifies significant PV stenosis by assessment of flow characteristics and vessel diameter and can thereby be used as a follow-up tool after catheter ablation of atrial fibrillation. PMID- 16697261 TI - Mitral valve ring dehiscence with an aorta-left atrial fistula. AB - In an era with the increasing use of various imaging modalities including echocardiography, ventriculography and cardiac magnetic resonance (CMR) imaging, one must be aware of the limitations of each discipline. We report a case of an individual who presented with both a partial dehiscence of a mitral valve annuloplasty ring and an aorta-left atrium fistula following surgical management of infective endocarditis that was correctly identified using transesophageal echocardiographic imaging. PMID- 16697262 TI - Subjective memory and mood of Hong Kong Chinese adults with epilepsy. AB - Clinicians are often confronted with the self-report of memory difficulties by patients. This study explored the possible correlates of subjective memory in 67 adult Chinese patients with epilepsy in Hong Kong. These correlates include epilepsy-related factors, mood state, and actual performance on neuropsychological tests. Results suggested that there exists no significant systematic relationship between subjective and illness-related factors such as seizure frequency, age at onset, and medication. Instead, stepwise regression analysis revealed that mood (anxiety) explained about 17% of the variance of subjective memory difficulties, whereas performance on a memory test accounted for only 8% of the variance. Findings are discussed in the light of the need to attend to the anxiety of patients with epilepsy in the process of rehabilitation. PMID- 16697263 TI - Psychiatric comorbidity in adult patients with hypothalamic hamartoma. AB - The objective was to investigate the prevalence of psychiatric comorbidity in adults with hypothalamic hamartoma (HH). A standardized clinical interview and assessment scales were used to assess for depression, anxiety, personality, and sexual behavior, based on previous findings in the pediatric population. We found a high prevalence of mood and anxiety disorders, with Major Depressive Disorder and Social Anxiety Disorder being the most common. "Anxious" personality types were the most common. Symptoms of Asperger syndrome were high in males versus females. We concluded that depression and anxiety are common in adult patients with HH. Remission of cognitive, behavioral, and autistic symptoms has been reported with surgical tumor resection. Whether this can correct the psychiatric disturbances remains to be investigated. The need for adjunctive psychiatric treatment may nevertheless remain. PMID- 16697264 TI - Psychogenic pseudosyncope: an underestimated and provable diagnosis. AB - OBJECTIVE: The goal of this study was to estimate the frequency of psychogenic pseudosyncope in patients with "syncope of unknown origin." Twenty to thirty percent of patients referred to epilepsy centers for refractory seizures have psychogenic seizures. With syncope, about 20-30% of the cases remain unexplained after a complete evaluation, but, unlike in seizures, a psychogenic etiology is not usually investigated. METHODS: We prospectively evaluated patients referred to our epilepsy center for evaluation of recurrent syncope-like episodes, that is, limp, motionless fainting. All patients had a negative syncope workup. We performed EEG-video monitoring with activation by suggestion ("induction"), similar to what is used for diagnosis of psychogenic seizures. Activation was performed with patients standing or sitting up. The diagnosis of psychogenic pseudosyncope required: (1) an activation procedure that triggered the habitual event; (2) a clinical event of loss of postural tone and limp, motionless unresponsiveness with eyes closed; (3) normal EEG before, during, and after the clinical event, that is, no epileptiform abnormalities, a normal alpha rhythm during unresponsiveness, and no suppression of background or slowing as is typically seen in syncope. RESULTS: Ten patients were recruited over an 18-month period. Habitual syncope-like episodes were triggered in 9 of 10 (90%) patients, and all 9 were shown to have psychogenic pseudosyncope (eyes closed, motionless, unresponsive with normal EEG including normal alpha rhythm). In one patient, no episode was triggered, so a diagnosis could not be made. Among the 9 patients for whom episodes were recorded, age ranged from 21 to 60 (mean=36). Five were women. Duration of symptoms ranged from 6 months to 15 years (mean=4.2 years). Event frequency ranged from four per day to two per month. Prior evaluations for syncope included ECG in all patients, two-dimensional echocardiogram in three, Holter monitoring in two, and tilt-table test in five. Four patients had undergone cardiac catheterization, and one had received a pacemaker. Neurologic tests included CT of the head in seven and MRI of the brain in eight. CONCLUSION: Many patients with "syncope of unknown origin" may have psychogenic pseudosyncope, but most such patients do not undergo EEG-video monitoring, which is the only way to demonstrate a psychogenic etiology. Psychogenic pseudosyncope is not simply a diagnosis of exclusion, and can be firmly diagnosed. As is usually recommended for seizure-like events, patients with syncope-like events and a negative evaluation should undergo EEG-video monitoring with induction, specifically looking for a possible psychogenic etiology. PMID- 16697265 TI - Intracellular cGMP may promote Ca2+-dependent and Ca2+-independent release of catecholamines from sympathetic nerve terminals. AB - OBJECTIVE: This study examined the hypothesis that intracellular cGMP stimulates the release of catecholamines from sympathetic nerve terminals (SNTs) in conscious rats. METHODS: Conscious rats were prepared to determine the effects of intravenously-administered agents on heart rate (HR) and mean arterial blood pressure (MAP). RESULTS: Bolus intravenous injections of the membrane-permeable cGMP analogue, 8-(4-chlorophenylthio)-cGMP (8-CPT-cGMP), elicited immediate and pronounced increases in HR before any changes in MAP were observed. In contrast, injections of cGMP did not elicit changes in HR or MAP. The 8-CPT-cGMP-induced tachycardia was markedly diminished by (1) the beta(1,2)-adrenoceptor antagonist, propranolol, (2) the ganglion blocking agent, chlorisondamine, and (3) bretylium, which blocks Ca2+-dependent mobilization of vesicular stores of catecholamines from SNTs. 8-CPT-cGMP also elicited minor falls in MAP in propranolol-treated rats but elicited pronounced falls in MAP in rats treated with chlorisondamine, bretylium, or combined administration of bretylium and the muscarinic receptor antagonist, methyl-atropine. CONCLUSIONS: These findings suggest that (1) intracellular cGMP elicits the release of Ca2+-sensitive and Ca2+-insensitive stores of catecholamines from SNTs in conscious rats, and (2) cGMP-mediated release of catecholamines from SNTs antagonizes cGMP-mediated relaxation of vascular smooth muscle in resistance arteries. Taken together, these findings support the concept that increases in intracellular cGMP levels by atrial natriuretic peptide and endothelium- and cardiac-derived nitric oxide regulate sympathetic control of the heart and the microvasculature of conscious rats via cGMP-dependent release of catecholamines. PMID- 16697266 TI - Effect of mangiferin on mitochondrial energy production in experimentally induced myocardial infarcted rats. AB - Mangiferin, from the leaves of Mangifera indica Linn., has been suggested as useful in the treatment of cardiovascular disorders. In the present study this drug was examined on the alteration of cardiac energy metabolism in isoproterenol (ISPH) administered myocardial infarcted rats. ISPH (20 mg/kg b.w.), which was administered s.c. twice at an interval of 24 h, caused a significant decrease in the activities of TCA cycle enzymes and antioxidant defense enzymes with a concomitant increase in the lipid peroxidation of heart mitochondria in rat model. The ATP production and the oxidation of succinate in State 3 and 4 decreased significantly in the cardiac mitochondria of ISPH administered rats. These functional alterations were supported by severe modifications in mitochondrial ultrastructure. Pretreatment with mangiferin (100 mg/kg b.w. i.p.) for 28 days prevented these mitochondrial alterations, oxidation with energy metabolism and restored the TCA cycle enzyme activities to near normal values following ISPH administration. The structural integrity of the heart was protected by mangiferin in ISPH administered rats when compared to the untreated controls. The present findings suggest that the protective effect of mangiferin can be attributed to its reducing effect on oxidative damage and activation of mitochondrial energy metabolism. These results could be useful to study and understand the cellular events involved in this cardioprotective mechanism of mangiferin. Our studies of mangiferin on heart failure may have important implication for future therapeutic approaches involving in the prevention of cardiovascular diseases. PMID- 16697267 TI - Effect of green tea on angiogenesis and severity of atherosclerosis in cholesterol-fed rabbit. AB - BACKGROUND AND AIMS: Since the development of the atherosclerotic plaque requires the growth of new microvessels in the plaque itself (vasa vasorum), we postulated that green tea may exert an anti-atherogenic effect. METHODS AND RESULTS: Thirteen male New Zealand white rabbits were studied for 17 weeks. All rabbits were fed an hypecholesterolemic diet. After 2 weeks of adaptation rabbits were randomly assigned into two groups. Animals in Group A were fed the hypercholesterolemic diet and received plain tap water ad libitum. Animals in Group B were fed with the same diet and furthermore received 2.5% (g/g) green tea for 17 weeks. CONCLUSION: According to our results the atherosclerotic lesions were more severe in Group B than in Group A specimens. Also, the number of VEGF positively stained foam cells and smooth muscle cells of Group B were significantly greater than in Group A. About 30% less plaque was found in Group A than in the control group (Group B). So, our study showed that the consumption of green tea leads to a reduction of atherosclerosis as well as a significant decrease of VEGF expression in the atherosclerotic plaque of rabbit aorta. The hypothesis that probably green tea may produce its anti-atherogenetic effect through an anti-angiogenetic mechanism needs more investigation. PMID- 16697268 TI - Nitric oxide synthases are involved in the modulation of cardiovascular adaptation in hemorrhaged rats. AB - AIM: Nitric oxide has been implicated in the cardiovascular adaptation to hemorrhagic shock. Our aim was to study the potential role of nitric oxide synthases (NOS) in the cardiovascular response in hemorrhagic hypotension produced experimentally in anesthetized rats. METHODS: Groups of animals (n = 14, per group): (a) normotensive; (b) hypovolemic (20% blood loss); (c) normotensive and pretreatment with N(G)-nitro-L-arginine methyl ester (L-NAME); (d) hypovolemic and pretreatment with L-NAME. RESULTS: L-NAME restored the hypotension induced by hemorrhage. Blood loss decreased heart rate in the first stage increasing at 60 and 120 min. L-NAME blunted this effect. Right atria and left ventricle histochemical NOS activities increased at 60 and 120 min (atria 8% and 24%, respectively; ventricle 21% and 45%, respectively). This activity increased 17% in smooth muscle at 120 min. Heart endothelial NOS protein levels increased in heart at 60 min being attenuated at 120 min. Inducible NOS protein levels raised significantly in right atria, left ventricle and aorta at 120 min. CONCLUSION: Hemorrhagic shock induced a time-dependent and specific NOS activation modulating cardiovascular function. The inhibition of nitric oxide system appears to prevent the acceleration of heart rate during late phases after acute hypovolemic state induced by blood loss. PMID- 16697270 TI - Right thoracic stomach resulting from hiatal hernia. PMID- 16697269 TI - The vasodilator potency of the endothelium-derived relaxing factor, L-S nitrosocysteine, is impaired in conscious spontaneously hypertensive rats. AB - OBJECTIVE: This study compared the hemodynamic responses elicited by the endothelium-derived relaxing factor (EDRF), L-S-nitrosocysteine (L-SNC), the non prostanoid EDRF released by acetylcholine (ACh) and nitric oxide (NO)-donors such as MAHMA NONOate, in conscious spontaneously hypertensive (SH) and normotensive Wistar-Kyoto (WKY) rats. METHODS: The depressor and/or vasodilator responses elicited by intravenous injections of ACh, L-SNC and MAHMA NONOate were determined in adult WKY and SH rats before and after intravenous injection of the NO synthesis inhibitor, N(G)-nitro-L-arginine methylester (L-NAME), or the cyclooxygenase inhibitor, indomethacin. RESULTS: The responses elicited by ACh and L-SNC were smaller in SH than in WKY rats whereas the responses elicited by MAHMA NONOate were augmented in SH rats. The ACh-induced responses were not diminished after injection of L-NAME in WKY or SH rats. Indomethacin did not affect the responses to any of the vasodilator agents in WKY or SH rats. Addition of L-SNC to whole blood or thoracic aortae from SH rats yielded similar amounts of NO to those of WKY rats. CONCLUSIONS: The vasodilator potencies of ACh and L SNC were diminished whereas that of NO was augmented in SH rats. The loss of potency of L-SNC in SH rats was not obviously due to differences in decomposition to NO or the overactivity of cyclooxygenase factors. This study provides the first evidence that diminished endothelium-dependent vasodilation in SH rats may involve a loss of vasodilator potency of endogenous L-SNC. PMID- 16697271 TI - Pneumatosis intestinalis and portal vein gas. PMID- 16697272 TI - Incidence of statin hepatotoxicity in patients with hepatitis C. AB - BACKGROUND AND AIMS: Statins are considered contraindicated in patients with chronic liver disease. Our objective was to determine the risk of developing hepatotoxicity from statin therapy in hyperlipidemic patients with hepatitis C. METHODS: Changes in liver biochemistry values within 12 months compared with baseline were determined in 3 cohorts matched for age, sex, and body mass index: (I) 166 anti-hepatitis C virus (HCV)-positive hyperlipidemic veterans who were initiated on statin therapy; (II) 332 anti-HCV-positive veterans who had not received statin therapy; and (III) 332 anti-HCV-negative hyperlipidemic veterans who were initiated on statin therapy. An increase in liver biochemistry values was defined as mild-moderate or severe as proposed in a previous study on statin hepatotoxicity in a non-hepatitis C population. RESULTS: In patients with hepatitis C, statin therapy (cohort I) was associated with a higher incidence of mild-moderate liver biochemistry value increases compared with those not on statin therapy (cohort II) (22.9% vs 13.3%, respectively, P = .009), but a lower incidence of severe increases (1.2% vs 6.6%, respectively, P = .015). Among patients started on statin therapy (cohorts I and III), the incidence of mild moderate liver biochemistry value increases (22.9% vs 16.3%, respectively, P = .094), severe increases (1.2% vs 1%, respectively, P = .874), or discontinuation of statin therapy as a result of hepatotoxicity (21.6% vs 9.2%, respectively, P = .147) were similar in hepatitis C-positive and hepatitis C-negative patients. CONCLUSIONS: Statin therapy was not associated with a higher risk of severe hepatotoxicity in patients with chronic hepatitis C and appeared safe. PMID- 16697273 TI - Tuberculous lymphadenitis: an uncommon cause of obstructive jaundice. PMID- 16697274 TI - Metastatic hepatic paraganglioma 6 years after carotid body tumor resection. PMID- 16697275 TI - Intestinal necrosis as a result of stanford type B aortic dissection. PMID- 16697276 TI - Munc13-1 is required for the sustained release of insulin from pancreatic beta cells. AB - Munc13-1 is a presynaptic protein that is essential for synaptic vesicle priming. Deletion of Munc13-1/unc13 causes total arrest of synaptic transmission due to a complete loss of fusion-competent synaptic vesicles. The requirement of Munc13-1 for large dense-core vesicles (LDCVs), however, has not been established. In the present study, we use Munc13-1 knockout (KO) and diacylglycerol (DAG) binding deficient Munc13-1(H567K) mutant knockin (KI) mice to determine the role of Munc13-1 in the secretion of insulin-containing LDCVs from primary cultured pancreatic beta cells. We show that Munc13-1 is required for the sustained insulin release upon prolonged stimulation. The sustained release involves signaling of DAG second messenger, since it is also reduced in KI mice. Insulin secretion in response to glucose stimulation is characterized by a biphasic time course. Our data show that Munc13-1 plays an essential role in the development of the second phase of insulin secretion by priming insulin-containing LDCVs. PMID- 16697277 TI - The plasma membrane redox system in aging. AB - Oxidative stress over time leads to the accumulation of damaged macromolecules and to profound physiological changes that are associated with several age related diseases. The plasma membrane redox system (PMRS) appears to attenuate oxidative stress acting as a compensatory mechanism during the aging process. The PMRS appears to play a protective role during mitochondrial dysfunction to provide cells with a survival mechanism by lowering oxidative stress. The PMRS accomplishes this by producing more NAD(+) for glycolytic ATP production via transfer of electrons from intracellular reducing equivalents to extracelluar acceptors. Ubiquinone and alpha-tocopherol are key antioxidant molecules in the plasma membrane that are affected by aging and can be up-regulated by dietary interventions such as calorie restriction (CR). Up-regulation of PMRS activity leads to cell survival and membrane homeostasis under stress conditions and during calorie restriction. Further studies of the PMRS may provide not only additional information on the mechanisms involved in aging and CR, but may provide therapeutic targets for the prevention and treatment of age-related diseases. PMID- 16697278 TI - Specific determination of intact cisplatin and monohydrated cisplatin in human plasma and culture medium ultrafiltrates using HPLC on-line with inductively coupled plasma mass spectrometry. AB - We have developed a specific assay for cisplatin in human plasma ultrafiltrate (PUF) and cell culture medium ultrafiltrate (MUF) using HPLC on-line with inductively coupled plasma mass spectrometry (ICP-MS). Separation of cisplatin (6 min) and monohydrated cisplatin (12 min) was achieved using a muBondapak C(18) column (Waters) and a mobile phase (0.075 mM sodium dodecyl sulfate and 3% methanol, adjusted to pH 2.5 with triflic acid) pumped at a flow rate of 0.5 mL/min. The analytes were detected with little background interference by ICP-MS monitoring of platinum masses (m/z 194/195). Calibration curves were linear over three orders of magnitude (0.05-8 microM) and the limit of quantitation was 0.1 microM. Intra- and inter-assay accuracy (range 91.6-113%) and precision (range 1.00-12.3%) were acceptable for PUF and MUF. The method was applied to determining cisplatin during ex vivo incubation of the drug in whole human blood at 37 degrees C. In conclusion, a specific, sensitive and reliable HPLC-ICP-MS assay has been established for determining intact cisplatin in PUF and MUF. PMID- 16697279 TI - Determination of omeprazole in rat plasma by high-performance liquid chromatography without solvent extraction. AB - A HPLC method without solvent extraction and using ultraviolet detection at 302 nm for the determination of omeprazole in rat plasma has been validated. Plasma samples after pretreatment with acetonitrile to effect deproteinization were dried under N(2) at 40 degrees C and reconstituted with mobile phase. The standard calibration curve for omeprazole was linear (r(2)=0.9999) over the concentration range of 0.02-3 microgml(-1). The intra- and inter-day assay variability range was 4.8-9.2% and 5.2-10.3% individually. This method has been successfully applied to a pharmacokinetic study of omeprazole in rats. PMID- 16697280 TI - Screening anion-exchange chromatographic matrices for isolation of onco retroviral vectors. AB - The adsorption kinetics of retroviral vectors to several chromatographic media, DEAE FF, Streamlinetrade mark Q XL and CHTtrade mark Ceramic Hydroxyapatite, in batch mode was investigated. The effects of buffer type, pH and operational temperature were studied. A mathematical model describing viral adsorption kinetics that considers viral degradation in solution was developed. The best results, either in terms of speed and extent of adsorbed infectious particles, were obtained with DEAE FF and Streamlinetrade mark Q XL. Fixed-bed chromatography was further investigated using DEAE FF, Q XL and Q FF, for validation of the batch adsorption process. Fixed-bed DEAE FF and Q XL proved to be good candidates for purification of MoMLV derived vectors due to resulting high yields, 53+/-13% and 51+/-7%, respectively, while removing more than 99% of protein and 90% of the DNA contaminants. PMID- 16697281 TI - Endogenous ligands of PACAP/VIP receptors in the autocrine-paracrine regulation of the adrenal gland. AB - Vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) are the main endogenous ligands of a class of G protein coupled receptors (Rs). Three subtypes of PACAP/VIP Rs have been identified and named PAC(1)-Rs, VPAC(1)-Rs, and VPAC(2)-Rs. The PAC(1)-R almost exclusively binds PACAP, while the other two subtypes bind with about equal efficiency VIP and PACAP. VIP, PACAP, and their receptors are widely distributed in the body tissues, including the adrenal gland. VIP and PACAP are synthesized in adrenomedullary chromaffin cells, and are released in the adrenal cortex and medulla by VIPergic and PACAPergic nerve fibers. PAC(1)-Rs are almost exclusively present in the adrenal medulla, while VPAC(1)-Rs and VPAC(2)-Rs are expressed in both the adrenal cortex and medulla. Evidence indicates that VIP and PACAP, acting via VPAC(1)-Rs and VPAC(2)-Rs coupled to adenylate cyclase (AC)- and phospholipase C (PLC)-dependent cascades, stimulate aldosterone secretion from zona glomerulosa (ZG) cells. There is also proof that they can also enhance aldosterone secretion indirectly, by eliciting the release from medullary chromaffin cells of catecholamines and adrenocorticotropic hormone (ACTH), which in turn may act on the cortical cells in a paracrine manner. The involvement of VIP and PACAP in the regulation of glucocorticoid secretion from inner adrenocortical cells is doubtful and surely of minor relevance. VIP and PACAP stimulate the synthesis and release of adrenomedullary catecholamines, and all three subtypes of PACAP/VIP Rs mediate this effect, PAC(1)-Rs being coupled to AC, VPAC(1)-Rs to both AC and PLC, and VPAC(2)-Rs only to PLC. A privotal role in the catecholamine secretagogue action of VIP and PACAP is played by Ca(2+). VIP and PACAP may also modulate the growth of the adrenal cortex and medulla. The concentrations attained by VIP and PACAP in the blood rule out the possibility that they act as true circulating hormones. Conversely, their adrenal content is consistent with a local autocrine-paracrine mechanism of action. PMID- 16697282 TI - Ultrastructural dynamics of human reproduction, from ovulation to fertilization and early embryo development. AB - This study describes the updated, fine structure of human gametes, the human fertilization process, and human embryos, mainly derived from assisted reproductive technology (ART). As clearly shown, the ultrastructure of human reproduction is a peculiar multistep process, which differs in part from that of other mammalian models, having some unique features. Particular attention has been devoted to the (1) sperm ultrastructure, likely "Tygerberg (Kruger) strict morphology criteria"; (2) mature oocyte, in which the MII spindle is barrel shaped, anastral, and lacking centrioles; (3) three-dimensional microarchitecture of the zona pellucida with its unique supramolecular filamentous organization; (4) sperm-egg interactions with the peculiarity of the sperm centrosome that activates the egg and organizes the sperm aster and mitotic spindles of the embryo; and (5) presence of viable cumulus cells whose metabolic activity is closely related to egg and embryo behavior in in vitro as well as in vivo conditions, in a sort of extraovarian "microfollicular unit." Even if the ultrastructural morphodynamic features of human fertilization are well understood, our knowledge about in vivo fertilization is still very limited and the complex sequence of in vivo biological steps involved in human reproduction is only partially reproduced in current ART procedures. PMID- 16697283 TI - Chromosomal variation in mammalian neuronal cells: known facts and attractive hypotheses. AB - Chromosomal mosaicism is still a genetic enigma. Although the mechanisms and consequences of this phenomenon have been studied for over 50 years, there are a number of gaps in our knowledge concerning causes, genetic mechanisms, and phenotypic manifestations of chromosomal mosaicism. Neuronal cell-specific chromosomal mosaicism is not an exception. Originally, neuronal cells of the mammalian brain were assumed to possess identical genomes. However, recent studies have shown chromosomal variations, manifested as chromosome abnormalities in cells of the developing and adult mammalian nervous system. Here, we review data obtained on the variation in chromosome complement in mammalian neuronal cells and hypothesize about the possible relevance of large-scale genomic (i.e., chromosomal) variations to brain development and functions as well as neurodevelopmental and neurodegenerative disorders. We propose to cover the term "molecular neurocytogenetics to cover all studies the aim of which is to reveal chromosome variations and organization in the mammalian brain. PMID- 16697284 TI - Automated interpretation of protein subcellular location patterns. AB - Proteomics is a major current focus of biomedical research, and location proteomics is the important branch of proteomics that systematically studies the subcellular distributions for all proteins expressed in a given cell type. Fluorescence microscopy of labeled proteins is currently the main methodology to obtain location information. Traditionally, microscope images are analyzed by visual inspection, which suffers from inefficiency and inconsistency. Automated and objective interpretation approaches are therefore needed for location proteomics. In this article, we briefly review recent advances in automated imaging interpretation tools, including supervised classification (which assigns location pattern labels to previously unseen images), unsupervised clustering (which groups proteins based on the similarity among their subcellular distributions), and additional statistical tools that can aid cell and molecular biologists who use microscopy in their work. PMID- 16697285 TI - Cell and molecular biology of human lacrimal gland and nasolacrimal duct mucins. AB - The old concept that the lacrimal gland is only a serous gland has been superseded by the finding that lacrimal acinar cells are able to produce mucins- high-molecular-weight proteins--the major mass being carbohydrates with the common feature of tandem repeats of amino acids rich in serine, threonine, and proline in the central domain of the mucin core peptide. At the ocular surface, maintenance of the tear film, lubrication, and provision of a pathogen barrier on the epithelia, conjunctiva, and cornea have been shown to be facilitated by mucins that are present in membrane-anchored (lining epithelial cells) or secreted (goblet cells) form. Also in the lacrimal gland, both membrane-anchored (MUCs 1, 4, and 16) and secreted (MUCs 5B and 7) mucins have been identified. The lacrimal gland is the main contributor to the aqueous portion of the tear film. It is part of the lacrimal apparatus that comprises, together with the lacrimal gland, the paired lacrimal canaliculi, the lacrimal sac, and the nasolacrimal duct, which collects the tear fluid and conveys it into the nasal cavity. In this review, the latest information regarding mucin function in the human lacrimal gland and the human efferent tear ducts is summarized with regard to mucous epithelia integrity, rheological and antimicrobial properties of the tear film and tear outflow, age-related changes, and certain disease states such as the pathogenesis of dry eye, dacryostenosis, and dacryolith formation. PMID- 16697288 TI - Classification matters for catatonia and autism in children. AB - Despite its chequered history, Kahlbaum's 1874 description of catatonia (tension insanity) and its categorization as a clinical illness is in outline still valid. Kahlbaum also acknowledged the existence of catatonia in children. Corresponding case studies have also been analyzed. The originators and disciples of the Wernicke-Kleist-Leonhard school proved catatonia in early childhood as a discrete entity with specific psychopathology. This does not mean that catatonic symptoms do not occur in other illnesses and in particular in organic psychoses. These are, however, of a totally different nature. Autism, as first described in connection with schizophrenic negativism by Bleuler in 1910, is one of the key symptoms of schizophrenia. As identified by Kanner in 1943, abnormal social interaction and communication, together with retarded development, are the main characteristics of autism in early childhood. Asperger's concept of autistic disorder (1944), although based on psychopathological theory, did not include retardation in development as an aspect. Consequently, autistic behavior can occur in a variety of mental disorders. Research into possible etiological and pathogenetic factors has been undertaken, but no clear link found as yet. PMID- 16697289 TI - A systematic examination of catatonia-like clinical pictures in autism spectrum disorders. AB - Three studies concerning catatonia-like clinical pictures in people with autism spectrum disorders (ASDs) referred to clinics are described. The first investigated the frequencies, in children and adults with autistic disorders, of 28 specific disorders of movement, speech, and behavior similar to those occurring in chronic catatonia spectrum conditions. The second compared the frequency of these items among groups of children with, ASDs, learning disabilities, specific language impairment, and a group with typical development, respectively. The third study examined the pattern of catatonia-like deterioration occurring in a minority of adolescents and adults with ASDs. The studies demonstrated the high frequency of catatonia-like features in people with autistic disorders. There was some tendency for improvement with increasing age, especially for those with IQ 70 or over. The items were also found in children with learning disabilities and specific language disorders but significantly less often. They occurred least often in the children with typical development. Severe catatonia-like deterioration occurred in 17% of those with autistic disorders, who were aged 15 years or over when assessed at a diagnostic center. A history of passivity in social interaction and impairment of expressive language were associated with the deterioration. No clear relationship was found between a history of catatonia-like features, singly or combined, and catatonia-like deterioration. The findings pose questions for future research. PMID- 16697290 TI - Catatonia in individuals with autism spectrum disorders in adolescence and early adulthood: a long-term prospective study. AB - The objective is to cast light on diagnosis and catastasis, course, and comorbidity as concerned with catatonia in patients with autism spectrum disorders (ASDs) with respect to long-term prospective follow-up. Eleven patients (all male) were enrolled. The mean age and the mean follow-up duration were 27.6 years (standard deviation (SD) 5.5) and 18.7 years (SD 8.7), respectively. The mean IQ was 27 (SD 16.4). Information was garnered from medical case records; current examination and observation of patients, interview of parents, and questionnaires completed by parents or other caretakers. Informed consent was obtained from the parents. Criteria for catatonia in this study were: (1) abrupt stop of movements and maintenance of immobility or bizarre posture beginning in adolescence and early adult life, (2) such a cataleptic state had continued for at least several minutes and appeared many times a day to the point of interfering with daily activities. We described two typical catatonic cases of ASDs. The average onset age was 19 years (SD 6). In all cases, our diagnostic criteria of catatonia evaluating at worse are fully compatible with those of Diagnostic and Statistical Manual of Mental Disorders, 4th ed. (DSM-VI). In 8 out of 11, the onset of catatonia was clearly preceded by the appearance of slowness in movements accompanying the exacerbation of obsessive-compulsive symptoms. Catatonia was also found to have some connection with Tourette syndrome (3 cases), adjustment disorders (N=1), and depressive mood disorders (N=1). In one case, the manifestations of catatonia had to be distinguished from parkinsonism caused by antipsychotics. Catatonia in ASDs seems to be a chronic condition in most cases. However, there were also a few cases in which catatonia repeatedly aggravated over short spans of time. Catatonia in ASDs may be considered an epiphenomenon of ASDs or a manifestation of comorbidity in adolescence or early adulthood. PMID- 16697291 TI - Are autistic and catatonic regression related? A few working hypotheses involving gaba, Purkinje cell survival, neurogenesis, and ECT. AB - Autistic regression seems to occur in about a quarter of children with autism. Its cause is unknown. Late-onset autistic regression, that is, after 2 years of age, shares some features with catatonic regression. A working hypothesis is developed that some children with autistic regression suffer from early-onset catatonic regression. This hypothesis cannot be answered from current data and is difficult to address in clinical studies in the absence of definite markers of autistic and catatonic regression. Treatment implications are theoretical and involve the potential use of anticatatonic treatments for autistic regression. Focus is on electroconvulsive therapy (ECT)--an established but controversial treatment that is viewed by many, but not all, as the most effective treatment for severe, life-threatening catatonic regression. Clinical trials of ECT in early- or late-onset autistic regression in children have not been done yet. The effects of electroconvulsive seizures--the experimental analogue of ECT--should also be tested in gamma-aminobutyric acid-ergic animal models of autistic regression, autism, catatonia, and other neurodevelopmental disorders. Purkinje cell survival and neurogenesis are putative outcome measures in these models. PMID- 16697292 TI - Psychomotor development and psychopathology in childhood. AB - The sensorimotor developmental phase, leading to a gradual acquisition of skilled actions, is of crucial importance for the young child and its growing sense of competence. Three vital steps in motor development are mentioned: first, the smooth and spontaneous movements of the "graceful and elegant" baby, expression of his well-being and vitality, with their profound effect on the mother-infant relationship; second, the emergence of intentional and goal-oriented acts leading to Funktionslust and playful repetitions; and finally, the development of symbolic acts and increasing technical capacity to use playthings in imaginative play. The psychodynamic significance of the most important motor milestones for the child's ego development is set out. Motility is one of the most important avenues for exercising such functions as mastery, integration, reality testing (self-preservation), and control of impulses. One may consider this early childhood period of rapid motor development as the motor phase of ego and libido development. Hence, many forms of developmental psychopathology are attended with motor impairment or insufficient motor mastery and integration. From that clinical perspective pass in review: perinatal complications and motor disturbance, attention deficit/hyperactivity disorder, dissociated motor development, low birth weight children and their developmental difficulties, developmental coordination disorder, aspects of pervasive developmental disorder, and stereotypic movement disorder. PMID- 16697293 TI - The importance of catatonia and stereotypies in autistic spectrum disorders. AB - Motor disturbances are often observed in individuals with autistic spectrum disorders (ASDs) and recognized as diagnostic features of these disorders. The movement disorders characteristically associated with autism include stereotypies and self-injurious behavior. Yet, individuals with ASD may also be at the risk for catatonia. Although not as frequent as stereotypies, up to 17% of older adolescents and adults with autistic disorder may have severe catatonic-like symptoms. Catatonia may be a comorbid risk factor of autism that warrants further empirical and clinical evaluations. Clinicians may need to be attentive to more subtle signs of catatonic-like symptoms in individuals diagnosed with ASDs, especially as they enter adolescence and young adulthood. Stress has been implicated as a possible precursor for symptoms; however, its role has not been empirically proven as a potential risk factor. Clinicians might also need to assess for signs of significant declines in motor movements, as this appears to be a useful diagnostic indicator of catatonic-like symptoms. The literature on stereotypies and autism is more extensive than for catatonia and ASDs, probably because of the higher rate of stereotypies with autism. Explanations for the occurrence of stereotypies range from genetic to behavioral contingencies, with evidence for a multifactor explanation. Assessment measures often include items that assess for stereotypies to aid with diagnosing these symptoms in individuals with autism. Treatment for stereotypies is largely behavioral at the present time and requires consistent reinforcement of treatment gains to manage the symptoms successfully. An important area of future research in autism is the relation among different types of motor abnormalities, including stereotypies and catatonia. PMID- 16697294 TI - Prader-Willi syndrome: atypical psychoses and motor dysfunctions. AB - Prader-Willi syndrome (PWS) is the result of a lack of expression of genes on the paternally derived chromosome 15q11-q13 and can be considered as a hypothalamic disorder. Its behavioral phenotype is characterized by ritualistic, stereotyped, and compulsive behaviors as well as motor abnormalities. After adolescence, recurrent affective psychoses are relatively frequent, especially in patients with uniparental disomy. These psychotic states have a subacute onset with complete recovery and comprise an increase of psychomotor symptoms that show resemblance with catatonia. Some evidence has emerged that gamma-aminobutyric acid (GABA) dysfunctionality is involved in both PWS and catatonia. Treatment of these atypical psychoses should preferably include GABA mimetic compounds like lorazepam, valproic acid, and possibly topiramate. PMID- 16697295 TI - Towards a valid nosography and psychopathology of catatonia in children and adolescents. AB - Paraphrasing Taylor and Fink (2003), catatonia needs "a home of its own" in child and adolescent psychiatry. Limited but expanding literature supports that catatonia in children and adolescent can be identified reliably among other childhood conditions, is sufficiently common, treatable with the same specific treatments as adult catatonia (e.g., sedative drugs and electroconvulsive therapy), and can be worsened by other treatments (e.g., antipsychotics). Other findings in child and adolescent catatonia suggest that sex ratio and associated disorders may differ, and the proposed classification of Taylor and Fink (2003) needs modification. Adopting a broader diagnostic schedule may accommodate both child, adolescent, and adult catatonia. A psychomotor automatism variant should be included as a diagnosis, as well as specifiers for associated disorders such as acute nonpsychotic anxious state and pervasive developmental disorder. Duration of illness should be specified as acute or chronic. Regardless of associated psychiatric disorders, this chapter describes a new psychopathological model. Three main modalities of movement dysfunction in catatonic subjects are listed: (1) adherence to delusional ideas leading to a psychomotor automatism (De Clerambault, 1927); (2) resistance to delusional thinking or conviction; and finally (3) hyperanxious states. Case-vignettes illustrate the model, and future research directions are identified. PMID- 16697296 TI - Is there a common neuronal basis for autism and catatonia? AB - Neuronal bases for autism and catatonia are unknown although integrative theories may soon become feasible as research in autism and catatonia advances. Catatonia and autism may both qualify as neurobiological syndromes in their own right. There is emerging evidence that catatonia may be a common syndrome in autism. Although the relation between autism and catatonia is unclear, coexpression of autism and catatonia may be due to abnormalities in common neuronal circuitries. This possibility constitutes another level of complexity to neurobiological inquiry, but also provides an opportunity to advance our understanding of both disorders. There is a great potential benefit in studying the relation between catatonia and autism in order to focus future research on subtype-specific causes and treatments. Future research avenues are outlined. PMID- 16697297 TI - Shared susceptibility region on chromosome 15 between autism and catatonia. AB - We have compiled significant linkage results from 20 genome scans for the autism syndrome disorder (ASD) and 2 for catatonia in schizophrenia (SZ). Localization of the markers has been updated across the studies using the same cytological (Genetic Location Database), physical (National Center for Biological Information), and genetic (Marshfield) maps. Eight autosomal chromosomes (1, 2, 3, 7, 9, 13, 15, and 17) showed significant linkages with ASD, and one with catatonia (15). Chromosome 15 was further characterized for SZ genome scans (N = 4) since catatonia was observed in SZ patients, for candidate genes for ASD and catatonia, and for the numerous chromosomal rearrangement and abnormalities associated to ASD. From these results, we observed that four potential susceptibility regions for ASD could be observed on chromosome 15 at 15q11-q13, 15q14-q21, 15q22-q23, and 15q26, respectively. All the four regions were shared between ASD and SZ, with 15q15-q21 being also shared with catatonia. Strong candidate genes, such as gamma-aminobutyric acid receptor B3, A5, and G3, have shown associations with ASD at 15q11-q13 susceptibility region where the majority of the chromosomal rearrangements are also found. On the other hand, negative association results were observed at 15q14-q21 susceptibility region for catatonia with the genes encoding the zinc transporter SLC30A4, the cholinergic receptor nicotinic alpha polypeptide 7, and the delta-like 4 Drosophila. Further, fine mapping and candidate gene analyses are needed to highlight potential common genes between ASD and catatonia for this chromosome. PMID- 16697298 TI - Current trends in behavioral interventions for children with autism. AB - This chapter focuses on behavioral interventions involving skill acquisition for children and adolescents with autism over the last 12 years. Literature and outcome data are reviewed with respect to three categories: language, social skills, and self-management. Generalization of results is somewhat problematic, as all of the interventions discussed consist of relatively small sample sizes and utilize single case design methodology. However, expansion and refinement of research methodology over the last decade, with more researchers replicating previous studies, may make broader application of research findings possible. PMID- 16697299 TI - Case reports with a child psychiatric exploration of catatonia, autism, and delirium. AB - This chapter starts with some remarks on the conceptual history of catatonia, which begins with Kahlbaum and continues with Kraeplin, Bleuler, and Leonhard. The Diagnostic and Statistical Manual, 4th ed., Text Revision, criteria for catatonia and the multicausal origin of the disorder are discussed. So, not only schizophrenia and mood disorders associated with catatonia, which is the primary form, are introduced but also an extensive list of somatic disorders-resulting in secondary catatonia-along with the work of Gelenberg and Wing. Next, two very difficult cases, of boys with autism, catatonia, and one of them with mental retardation as well, are presented. Major textbooks, PubMed, and Medline were used for a select literature search. The results show the main and really relevant but scarce data concerning primary and secondary catatonia. In the discussion the topics are this dearth in knowledge, the concept of catatonia and its similarities with delirium, and the relation catatonia-autism, and where to find the data. The conclusions summarize the main points and end with a gentle reminder, or is it an appeal? PMID- 16697300 TI - ECT and the youth: catatonia in context. AB - Electroconvulsive therapy (ECT) has been in psychiatric practice for well over half a century, but it continues to incite controversy. However, it is regarded amongst psychiatrists as a safe and effective treatment and at times even a lifesaver. It offers a fairly swift but a time-limited response, opening up opportunities for initiation of more longer lasting treatments. The use of ECT in the youth is limited, and as such good studies are few and far between. The recent Practice Parameters by the American Academy of Child and Adolescent Psychiatry, specifically addressing ECT in adolescents, is indeed a welcome addition. Electrocovulsive therapy is as effective in the youth as it is in the adults, and the indications and contraindication are the same. The administration of ECT follows the same general principles in all age groups. One particular indication is of the use in catatonia, a motor syndrome that could occur with affective disorders, schizophrenia or medical conditions, in which it is considered to be extremely effective. The association between catatonia and autism and spectrum disorders has been noted, and in this situation, ECT is considered by some to be effective. Ethical considerations and that of capacity and informed consent are of paramount importance as are the human rights. Working in partnership with the parents/carers all the way is a must. The lack of information leaflets on ECT especially designed for young patients and their parents has to be rectified soon. Registers based on geographical health regions for those below the age of 18 will assist tremendously in epidemiological studies as well as pave the way toward more evidence-based studies that are essential. PMID- 16697301 TI - Catatonia in autistic spectrum disorders: a medical treatment algorithm. AB - Autism is a developmental syndrome with an unknown biology and inadequate therapeutics. Assessing the elements of the syndrome for the presence of depression, psychosis, mania, or catatonia, offers opportunities for systematic intervention. Since almost all descriptions of autism highlight the presence of motor symptoms that characterize catatonia, an assessment for this eminently treatable syndrome is recommended for all patients considered to be autistic. A minimum examination includes a catatonia rating scale and for those patients with defined catatonia, a lorazepam test. For those whose catatonia responds to lorazepam, high dose lorazepam therapy is recommended. If this fails, electroconvulsive therapy is recommended. The assessment and treatment of catatonia offers positive medical therapy for the victims of autism and their families. PMID- 16697302 TI - Psychological approaches to chronic catatonia-like deterioration in autism spectrum disorders. AB - The psychological dysfunctions that may underlie catatonia-like deterioration in autism spectrum disorders are discussed. Clinical observation suggests that an important factor is ongoing stress. The evidence for this from research and clinical observation is considered. The lack of evidence concerning the most appropriate medical treatments is discussed. A psychological approach designed for individual needs by relevant professionals and applied by parents and/or caregivers is described. This can be helpful whether or not medical treatments are used. It involves detailed holistic assessment of the individual and their circumstances to highlight possible precipitating stress factors in view of their underlying autism and cognitive/psychological functioning. The overall aim of this approach is to restructure the individual's lifestyle, environment and resolve cognitive/psychological factors to reduce the stress. An eclectic approach is used to find individual strategies in order to provide external goals and stimulation to increase motivation and keep the person engaged and active in meaningful and enjoyable pursuits. The approach describes ways of using verbal and physical prompts as external stimuli to overcome the movement difficulties and emphasizes maintaining a predictable structure and routine for each day. The importance of educating caregivers and service providers to understand the catatonia-like behavior is emphasized. Advice is given on management of specific problems such as incontinence, freezing in postures, eating problems, and episodes of excitement. PMID- 16697303 TI - Blueprints for the assessment, treatment, and future study of catatonia in autism spectrum disorders. AB - The blueprints for the assessment, treatment, and future study of catatonia in autism spectrum disorders (ASDs), which are submitted in this chapter aim to increase early recognition and treatment of catatonia in ASDs, show the urgency of controlled treatment trials, and increase collaborative and interdisciplinary research into the co-occurrence of these two enigmatic disorders. Catatonia should be assessed in any patient with ASDs when there is an obvious and marked deterioration in movement, pattern of activities, self-care, and practical skills, compared with previous levels, through a comprehensive diagnostic evaluation of medical and psychiatric symptoms. A formal diagnosis should be ascertained using ASD specific criteria for catatonia that takes into account baseline symptoms like muteness, echophenomena, stereotypy, negativism, or other psychomotor abnormalities. Any underlying medical and neurological conditions should be treated, and culprit medications or other substances that may cause catatonia should be eliminated. Separate treatment blueprints are presented for mild, moderate, and severe catatonia, featuring combinations of a psychological approach developed by Shah and Wing and medical treatments that have shown efficacy in catatonia: lorazepam challenge, lorazepam trial, lorazepam continuation, and bilateral electroconvulsive therapy (ECT). These treatment modalities in themselves are well established. Side effects and complications are known and manageable. Legal, ethical, and practice guidelines governing all treatment aspects should be followed. The treatment blueprints should be viewed as best estimates pending future controlled studies. The blueprint for the future study of catatonia in ASDs describes promising clinical and preclinical research avenues. Longitudinal studies need to assess the possible effect of early recognition and adequate treatment of catatonia in ASDs in order to avoid the impairment associated with chronicity. Effects of current and new anticatatonic treatments should be examined in experimental models of autism and catatonia. Finally, the role of gamma-aminobutyric acid (GABA) dysfunction in autism, catatonia, and abnormal stress responses in these disorders should be further assessed. PMID- 16697304 TI - Endocardial fibrosis in subacute non-Q wave myocardial infarction demonstrated by multislice computed tomography. PMID- 16697305 TI - Prospective evaluation of hospital isolation room capacity. AB - Risk assessment is used to determine the need for isolation in single rooms. Limited availability of isolation rooms and/or operational needs may compromise this process. This article reports the results of a 12-month prospective observational study of every infection control request for isolation in a 1100 bed teaching hospital. In addition, four point-prevalence surveys of the usage of single rooms were carried out. Data were collected on the incidence of new clinical meticillin-resistant Staphylococcus aureus (MRSA) isolates per ward and these were correlated with rates of isolation failures for MRSA cases. There were 845 requirements for patient isolation, of which 185 (22%) could not be met (isolation failures). Three-quarters of the requirements for isolation were due to MRSA or Clostridium difficile. The proportion of isolation failures was consistent for most organisms and conditions but varied markedly between clinical specialities (0-57%). Reasons for failure to isolate included no single rooms available, all single/isolation rooms occupied (for both isolation and non infection-control reasons), limitations on the use of single rooms in mixed-sex wards and patient-specific reasons. Only a minority of the available single rooms were occupied for infection control reasons (12-19%). There was a statistically significant correlation between isolation failures and MRSA incidence (Spearman's rho 0.596, P<0.001). In only one case where a ward had >or=30% of its beds provided in single rooms was there an instance of failure to isolate. In conclusion, insufficient capacity to isolate patients with potentially transmissible pathogens is common and may compromise infection control requirements. Either isolation capacity must be increased or evidence-based risk assessment must be applied to situations where demand for isolation exceeds availability. Further information is needed on the consequences of isolation failure. PMID- 16697306 TI - Rimonabant: a cannabinoid receptor type 1 blocker for management of multiple cardiometabolic risk factors. AB - Rimonabant is a first selective blocker of the cannabinoid receptor type 1 (CB1) being developed for the treatment of multiple cardiometabolic risk factors, including abdominal obesity and smoking. In four large trials, after one year of treatment, rimonabant 20 mg led to greater weight loss and reduction in waist circumference compared with placebo. Therapy with rimonabant is also associated with favorable changes in serum lipid levels and an improvement in glycemic control in prediabetes patients and in type 2 diabetic patients. At the same dose, rimonabant significantly increased cigarette smoking quit rates as compared with placebo. Rimonabant seems to be well tolerated, with a primary side effect of mild nausea. As an agent with a novel mechanism of action, rimonabant has a potential to be a useful adjunct to lifestyle and behavior modification in treatment of multiple cardiometabolic risk factors, including abdominal obesity and smoking. PMID- 16697307 TI - Biventricular versus conventional right ventricular stimulation for patients with standard pacing indication and left ventricular dysfunction: the Homburg Biventricular Pacing Evaluation (HOBIPACE). AB - OBJECTIVES: The Homburg Biventricular Pacing Evaluation (HOBIPACE) is the first randomized controlled study that compares the biventricular (BV) pacing approach with conventional right ventricular (RV) pacing in patients with left ventricular (LV) dysfunction and a standard indication for antibradycardia pacing in the ventricle. BACKGROUND: In patients with LV dysfunction and atrioventricular block, conventional RV pacing may yield a detrimental effect on LV function. METHODS: Thirty patients with standard indication for permanent ventricular pacing and LV dysfunction defined by an LV end-diastolic diameter > or =60 mm and an ejection fraction < or =40% were included. Using a prospective, randomized crossover design, three months of RV pacing were compared with three months of BV pacing with regard to LV function, N-terminal pro-B-type natriuretic peptide (NT proBNP) serum concentration, exercise capacity, and quality of life. RESULTS: When compared with RV pacing, BV stimulation reduced LV end-diastolic (-9.0%, p = 0.022) and end-systolic volumes (-16.9%, p < 0.001), NT-proBNP level (-31.0%, p < 0.002), and the Minnesota Living with Heart Failure score (-18.9%, p = 0.01). Left ventricular ejection fraction (+22.1%), peak oxygen consumption (+12.0%), oxygen uptake at the ventilatory threshold (+12.5%), and peak circulatory power (+21.0%) were higher (p < 0.0002) with BV pacing. The benefit of BV over RV pacing was similar for patients with (n = 9) and without (n = 21) atrial fibrillation. Right ventricular function was not affected by BV pacing. CONCLUSIONS: In patients with LV dysfunction who need permanent ventricular pacing support, BV stimulation is superior to conventional RV pacing with regard to LV function, quality of life, and maximal as well as submaximal exercise capacity. PMID- 16697308 TI - Prevention of ventricular desynchronization by permanent para-Hisian pacing after atrioventricular node ablation in chronic atrial fibrillation: a crossover, blinded, randomized study versus apical right ventricular pacing. AB - OBJECTIVES: The aim of our study was to evaluate the feasibility, the safety, and hemodynamic improvements induced by permanent para-Hisian pacing in patients with chronic atrial fibrillation and narrow QRS who underwent atrioventricular (AV) node ablation. BACKGROUND: Right ventricular apical pacing, inducing asynchronous ventricular contraction, may impair cardiac function; permanent para-Hisian pacing could preserve interventricular synchrony and improve left ventricular function. METHODS: After AV node ablation, 16 patients were implanted with a dual chamber pacemaker connected to a screw-in lead positioned in close proximity to the His bundle and to a right ventricular apical lead. Clinical and echocardiographic data were collected at baseline and after two randomized six month periods (with para-Hisian and conventional pacing). RESULTS: During para Hisian pacing, the interventricular electromechanical delay improved as well (34 +/- 18 ms) as during right apical pacing (47 +/- 19 ms), p < 0.05. Para-Hisian pacing allowed an improvement in New York Heart Association functional class (1.75 +/- 0.4 vs. 2.33 +/- 0.6 at baseline and 2.5 +/- 0.4 during apical pacing, p < 0.05 for both), in quality-of-life score (16.2 +/- 8.7 vs. 32.5 +/- 15.0 at baseline, p < 0.05), and in the 6-min walk test (431 +/- 73 m vs. 378 +/- 60 m at baseline and 360 +/- 71 m during apical pacing, p < 0.5 for both). Mitral and tricuspid regurgitation improved during para-Hisian pacing (1.22 +/- 0.8 and 1.46 +/- 0.5 index, respectively, vs. 1.68 +/- 0.6 [p < 0.05] and 1.62 +/- 0.7 [p = NS] index at baseline, respectively), with a slight worsening during apical pacing (1.93 +/- 1 and 1.93 +/- 0.7 index, respectively, p < 0.05 for both). CONCLUSIONS: Permanent para-Hisian pacing is feasible and safe. Compared with conventional right apical pacing, it allows an improvement in functional and hemodynamic parameters over long-term follow-up. PMID- 16697309 TI - Alternative site pacing to promote cardiac synchrony: has conventional pacing become unconventional? PMID- 16697310 TI - Long-term outcomes after stenting of bifurcation lesions with the "crush" technique: predictors of an adverse outcome. AB - OBJECTIVES: The purpose of this study was to evaluate predictors of an adverse outcome after "crush" bifurcation stenting. BACKGROUND: The "crush" technique is a recently introduced strategy with limited data regarding long-term outcomes. METHODS: We identified 231 consecutive patients treated with drug-eluting stent implantation with the "crush" technique for 241 de novo bifurcation lesions. Clinical follow-up was obtained in 99.6%. RESULTS: The in-hospital major adverse cardiac event (MACE) rate was 5.2%. At 9 months, 10 (4.3%) patients had an event consistent with possible post-procedural stent thrombosis. Survival free of target lesion revascularization (TLR) was 90.3%; the only independent predictor of TLR was left main stem (LMS) therapy (odds ratio [OR] 4.97; 95% confidence interval [CI] 2.00 to 12.37, p = 0.001). Survival free of MACE was 83.5% and independent predictors of MACE were LMS therapy (OR 3.79; 95% CI 1.76 to 8.14, p = 0.001) and treatment of patients with multivessel disease (OR 4.21; 95% CI 0.95 to 18.56, p = 0.058). Angiographic follow-up was obtained in 77% of lesions at 8.3 +/- 3.7 months. The mean late loss of the main vessel and side branch were 0.30 +/- 0.64 mm and 0.41 +/- 0.67 mm, respectively, with binary restenosis rates of 9.1% and 25.3%. Kissing balloon post-dilation significantly reduced the side branch late lumen loss (0.24 +/- 0.50 mm vs. 0.58 +/- 0.77 mm, p < 0.001). CONCLUSIONS: The crush technique of bifurcation stenting with drug-eluting stents is associated with favorable outcomes for most lesions; however, efficacy appears significantly reduced in LMS bifurcations, and further research is needed before the technique can be routinely recommended in this group. Furthermore, the incidence of possible stent thrombosis is of concern and requires further investigation. Kissing balloon post-dilatation is mandatory to reduce side branch restenosis. PMID- 16697311 TI - Genetic polymorphisms of platelet glycoprotein Ia and the risk for premature myocardial infarction: effects on the release of sCD40L during the acute phase of premature myocardial infarction. AB - OBJECTIVES: The aim of this research was to evaluate the effect of genetic polymorphisms C807T and G1648A of platelet glycoprotein Ia (GPIa), on the risk for myocardial infarction (MI) and on the release of soluble CD40 ligand (sCD40L) during the acute phase of MI and one year after the event. BACKGROUND: C807T and G1648A polymorphisms affect the density of GPIa on platelet surface, but their effect on the risk for MI and the release of sCD40L is unknown. METHODS: The study population consisted of 219 patients with premature MI and 389 controls. One year after the event, 67 patients and 232 controls were recalled for the follow-up study. RESULTS: The risk for MI in 807TT was 2.296 (95% confidence interval [CI]: 1.187 to 4.440) p < 0.05 versus CC + CT, 2.269 (95% CI: 1.085 to 4.745) p < 0.05 versus CC, and 2.135 (95% CI: 1.080 to 4.219) p < 0.05 versus CT. During the acute phase of MI, sCD40L was higher in 807CT + TT compared with 807CC (p < 0.01), an effect persisting after one year (p < 0.01). The carriage of 807T allele was an independent predictor for sCD40L during the acute phase of MI (beta = 9.442 [standard error (SE): 2.526], p = 0.001) and in the same patients one year later (beta = 8.282 [SE: 2.044], p = 0.001). In healthy individuals, 807T allele was associated with higher sCD40L levels compared with 807CC (p < 0.05), only among those with von Willebrand factor greater than or equal to median. CONCLUSIONS: Genetic polymorphism C807T increases the risk for premature MI. 807T allele is an independent predictor for sCD40L levels during the acute phase of premature MI as well as one year after the event, while it is associated with elevated sCD40L levels in healthy subjects, only in the presence of high von Willebrand levels. PMID- 16697312 TI - Relationship between cardiovascular risk factors and atherosclerotic disease burden measured by intravascular ultrasound. AB - OBJECTIVES: The goal of this study was to determine the relationship between established cardiovascular risk factors and the extent of coronary atherosclerotic plaque. BACKGROUND: Few data exist correlating cardiovascular risk factors with volumetric measurements of coronary atheroma burden in patients with coronary artery disease. METHODS: Clinical characteristics, quantitative coronary angiography, and intravascular ultrasound (IVUS) were evaluated in subjects enrolled in a study comparing atorvastatin and pravastatin. Plaque areas were measured at 1-mm intervals to compute atheroma volume. The percent of cross sections with an abnormal intimal thickness (>0.5 mm) was determined. Data on cardiovascular risk factors were collected. RESULTS: In 654 subjects, atheroma volume averaged 174.5 mm3 and percent atheroma volume 38.9%. Atherosclerosis was present in 81.2% of 25,897 cross sections. In univariate analysis, there was a strong association between diabetes, male gender, and a history of either prior revascularization or stroke with percent atheroma volume. Hypertension or prior myocardial infarction was also predictive of more severe disease. Low-density lipoprotein and C-reactive protein were not significant predictors of greater disease burden. In multivariate analysis, diabetes, male gender, and a history of a prior interventional procedure remained strong predictors of increased atheroma volume. History of stroke, non-Caucasian race, and smoking status remained significant. Although multiple measures of IVUS disease burden were worse in subjects with diabetes, angiographic stenosis severity was not different. CONCLUSIONS: Male gender, diabetes, and a history of prior revascularization are strong independent predictors of atherosclerotic burden in coronary disease patients. Many risk factors did not predict angiographic disease severity, suggesting different mechanisms drive stenosis development and atheroma accumulation. PMID- 16697313 TI - Heart disease risk determines menopausal age rather than the reverse. AB - OBJECTIVES: The purpose of this study was to investigate whether a harmful cardiovascular risk profile accelerates menopause. BACKGROUND: Women with an early menopause are at an increased risk of cardiovascular disease. Although increased cardiovascular risk has been proposed as consequence of menopause, the alternative hypothesis, that increased premenopausal cardiovascular risk promotes early menopause, needs to be examined. METHODS: We used data from the Framingham Heart Study cohort. This study started in 1948 and has followed up participants biennially since then. Women who were premenopausal at study entry and who reached natural menopause after at least two examination rounds were included in the study (n = 695). Premenopausal age-independent levels of serum total cholesterol, relative weight, blood pressure, and Framingham risk score were determined, as well as premenopausal changes in cholesterol, body weight, and blood pressure. RESULTS: A higher premenopausal serum total cholesterol level was statistically significantly associated with an earlier age at menopause, as were increases in total serum cholesterol, relative weight, and blood pressure in the premenopausal period. A decrease in total serum cholesterol during premenopause was statistically significantly associated with later age at menopause. Decreasing blood pressure was associated with a later menopausal age, but this association was not statistically significant. A decrease in relative weight was associated with a significant earlier age at menopause. Each 1% higher premenopausal Framingham risk score was associated with a decrease in menopausal age of 1.8 years (95% confidence interval -2.72 to -0.92). CONCLUSIONS: The findings support the view that heart disease risk determines age at menopause. This offers a novel explanation for the inconsistent findings on cardiovascular disease rate and its relationship to menopausal age and effects of hormone replacement therapy. PMID- 16697314 TI - Menopause and cardiovascular risk cause or consequence? PMID- 16697315 TI - Renal impairment and outcomes in heart failure: systematic review and meta analysis. AB - OBJECTIVES: We estimated the prevalence of renal impairment in heart failure (HF) patients and the magnitude of associated mortality risk using a systematic review of published studies. BACKGROUND: Renal impairment in HF patients is associated with excess mortality, although precise risk estimates are unclear. METHODS: A systematic search of MEDLINE (through May 2005) identified 16 studies characterizing the association between renal impairment and mortality in 80,098 hospitalized and non-hospitalized HF patients. All-cause mortality risks associated with any renal impairment (creatinine >1.0 mg/dl, creatinine clearance [CrCl] or estimated glomerular filtration rate [eGFR] <90 ml/min, or cystatin-C >1.03 mg/dl) and moderate to severe impairment (creatinine > or =1.5, CrCl or eGFR <53, or cystatin-C > or =1.56) were estimated using fixed-effects meta analysis. RESULTS: A total of 63% of patients had any renal impairment, and 29% had moderate to severe impairment. After follow-up > or =1 year, 38% of patients with any renal impairment and 51% with moderate to severe impairment died versus 24% without impairment. Adjusted all-cause mortality was increased for patients with any impairment (hazard ratio [HR] = 1.56; 95% confidence interval [CI] 1.53 to 1.60, p < 0.001) and moderate to severe impairment (HR = 2.31; 95% CI 2.18 to 2.44, p < 0.001). Mortality worsened incrementally across the range of renal function, with 15% (95% CI 14% to 17%) increased risk for every 0.5 mg/dl increase in creatinine and 7% (95% CI 4% to 10%) increased risk for every 10 ml/min decrease in eGFR. CONCLUSIONS: Renal impairment is common among HF patients and confers excess mortality. Renal function should be considered in risk stratification and evaluation of therapeutic strategies for HF patients. PMID- 16697316 TI - Atrial fibrillation and risk of clinical events in chronic heart failure with and without left ventricular systolic dysfunction: results from the Candesartan in Heart failure-Assessment of Reduction in Mortality and morbidity (CHARM) program. AB - OBJECTIVES: We assessed the risk of adverse cardiovascular (CV) outcomes associated with atrial fibrillation (AF) in the Candesartan in Heart failure Assessment of Reduction in Mortality and morbidity (CHARM) program, which enrolled patients with chronic heart failure (CHF) and a broad range of ejection fractions (EFs). BACKGROUND: Atrial fibrillation is associated with an increased risk of adverse CV outcomes in patients with CHF and reduced EF. The risk of AF in patients with CHF and preserved left ventricular ejection fraction (PEF) is unknown. METHODS: A total of 7,599 patients with symptomatic CHF were randomized to candesartan or placebo. Patients were divided by baseline EF (< or =40% or >40%) in low or preserved EF groups. Major outcomes were cardiovascular death or hospitalization for worsening heart failure, and all-cause mortality. Median follow-up was 37.7 months. RESULTS: A total of 670 (17%) patients in the low EF group and 478 (19%) in the PEF group had AF at baseline. Atrial fibrillation predicted a high risk of cardiovascular morbidity and mortality regardless of baseline EF. Patients with AF and low EF had the highest absolute risk for adverse CV outcomes. However, AF was associated with greater relative increased risk of the major outcomes in patients with PEF than in patients with low EF: hazard ratio 1.72 (95% confidence interval [CI] 1.45 to 2.06) versus 1.29 (95% CI 1.14 to 1.46), respectively. The same was true for the risk of all-cause mortality. Candesartan was associated with similar treatment effects regardless of baseline rhythm. CONCLUSIONS: Atrial fibrillation is associated with an increased risk of CV outcomes in patients with CHF and either reduced EF or PEF. Candesartan improved outcomes similarly regardless of baseline rhythm. PMID- 16697317 TI - Sites of focal atrial activity characterized by endocardial mapping during atrial fibrillation. AB - OBJECTIVES: The aim of the present study was to assess the feasibility of identifying sites of focal atrial activity by localized high-density endocardial mapping during atrial fibrillation (AF). BACKGROUND: Sites of focal activity in the left atrium have been demonstrated by epicardial mapping during AF. METHODS: Twenty-four patients (15 with paroxysmal, 3 with persistent, and 6 with permanent AF) underwent endocardial mapping during AF. A 20-pole catheter with five radiating spines was used to map both atria for 30 s in each of 10 pre-determined segments. A focal activity was defined as > or =3 atrial cycles with activation spreading from center to periphery of the mapping catheter. Catheter ablation was performed independent of the mapping results. RESULTS: Spontaneous focal activities were observed in 13 sites in the left atrium (9%; anterior 1, roof 2, posterior 6, inferior 4) in 12 patients (9 paroxysmal, 3 persistent). Focal activity was observed continuously (two sites) or intermittently (11 sites, median 5 episodes), and associated with shortening of the cycle length (from 183 +/- 33 ms to 172 +/- 29 ms; p < 0.05). The mean duration of an intermittent episode was 1.5 s (range 0.4 to 7.1 s). Atrial fibrillation terminated without ablation at the foci in all of 12 patients, but in 2 of them, re-initiated arrhythmia was successfully ablated at these foci. Nine of these 12 patients (75%) were arrhythmia-free without antiarrhythmic drugs during a follow-up period of 7.0 +/- 3.1 months. CONCLUSIONS: Termination of AF without ablation at the sites of atrial focal activity suggests that this activity may be triggered by impulses originating from other regions, such as the pulmonary veins. PMID- 16697318 TI - Isolated potentials during sinus rhythm and pace-mapping within scars as guides for ablation of post-infarction ventricular tachycardia. AB - OBJECTIVES: The purpose of this study was to identify ventricular tachycardia (VT) isthmus sites by pace-mapping within scar tissue and to identify electrogram characteristics that are helpful in identifying VT isthmus sites during sinus rhythm (SR). BACKGROUND: Pace-mapping has been used in the scar border zone to identify the exit site of post-infarction VT. METHODS: In 19 consecutive patients (18 men, mean age 66 +/- 9 years, mean ejection fraction 0.24 +/- 0.12) with post infarction VT, a left ventricular voltage map was generated during SR. Pace mapping was performed at sites with abnormal electrograms or isolated potentials. Radiofrequency ablation was performed at isthmus sites as defined by pace-mapping (perfect pace-map = 12/12 matching electrocardiogram leads; good pace-map = 10/12 to 11/12 matching electrocardiogram leads) and/or entrainment mapping. RESULTS: A total of 81 VTs (mean cycle length 396 +/- 124 ms) were inducible. In 16 of the 19 patients, a total of 41 distinct isthmus areas of 41 distinct VTs were identified and successfully ablated. All but one displayed isolated potentials during SR. Furthermore, 22 of the 81 VTs (27%) for which no isthmus was identified became noninducible after ablation of a targeted VT. The 16 patients in whom > or =1 isthmus was identified and ablated were free of arrhythmic events during a mean follow-up of 10 months. CONCLUSIONS: During SR, excellent or good pace-maps at sites of isolated potentials within areas of scar identify areas of fixed block that are protected and part of the critical isthmus of post infarction VT. Shared common pathways might explain why non-targeted VTs might become noninducible after ablation of other VTs. PMID- 16697319 TI - Accuracy of multislice computed tomography in the preoperative assessment of coronary disease in patients with aortic valve stenosis. AB - OBJECTIVES: To evaluate multislice computed tomography (MSCT) as an alternative to coronary angiography, we prospectively studied its diagnostic accuracy for the detection of significant coronary artery lesions in patients with significant aortic valve stenosis undergoing valve surgery. BACKGROUND: In patients with aortic valve stenosis, coronary angiography is still recommended before surgery. Multislice computed tomography is a promising noninvasive technique for the detection of significant coronary artery lesions. METHODS: Fifty-five consecutive patients scheduled for coronary angiography in the preoperative assessment of aortic valve stenosis underwent 16-slice MSCT 24 h before coronary angiography. We analyzed coronary lesions, image quality, and arterial calcium score. RESULTS: The sensitivity of the MSCT-based strategy in detecting significant stenosis was 100%, and its specificity 80%. The positive and negative predictive values were respectively 55% and 100%. For calcium scores <1,000 (77% of patients), MSCT detected all patients without coronary artery disease, enabling conventional coronary angiography to be avoided in 35 of 55 cases (80%). For calcium scores >1,000, MSCT enabled conventional coronary angiography to be avoided in only 6% of cases, either because significant stenosis was found with a possible indication of revascularization, or because the examination was not interpretable. CONCLUSIONS: The results of this initial experience in relatively few patients suggest that MSCT-based coronary angiography may serve as an alternative to invasive coronary angiography to rule out significant coronary artery disease in patients scheduled for elective aortic valve replacement. Larger studies are necessary to fully explore the potential of coronary MSCT to improve preoperative risk stratification. PMID- 16697320 TI - Coronary multidetector computed tomography: a new standard for preoperative risk assessment? PMID- 16697321 TI - Effects of time, dose, and inversion time for acute myocardial infarct size measurements based on magnetic resonance imaging-delayed contrast enhancement. AB - OBJECTIVES: This study sought to investigate the influence of time, dose, and inversion time (TI) and their interactions on myocardial infarct size measurements to establish the foundation for a standardized protocol for multicenter trials. BACKGROUND: There is growing interest in using magnetic resonance imaging (MRI) infarct size measurements as an end point in clinical trials. However, no standardized protocol exists, and there are limited data concerning the effects of time, contrast agent dose, and TI. METHODS: First, we determined the influence of postcontrast imaging time (5 to 40 min), contrast agent dose (0.1 vs. 0.2 mmol/kg), TI, and their interactions in an animal model (n = 14). Second, we tested whether the findings of the animal study apply to patients and are generalizable. Therefore, we retested the diagnostic window in a multicenter study. A total of 48 patients with first acute myocardial infarction (AMI) from three centers were imaged twice (5 and 30 min) after injection of 0.15 mmol/kg gadolinium diethylenetriamine-pentaacetate using an adjusted TI. RESULTS: The animal study showed that the infarct size is independent of time and dose (p = 0.9 and p = 0.16, respectively) using an adjusted TI. Using a fixed TI, however, infarct size is a function of time and dose (p = 0.0001 and p = 0.01, respectively). The multicenter study showed that MRI 1 (16.9 +/- 12% of left ventricle) was not statistically different from MRI 2 (16.4 +/- 12% of left ventricle, p = NS) with no difference between sites (p = NS). CONCLUSIONS: The AMI size can be measured with MRI using a contrast dose between 0.1 and 0.2 mmol/kg and a time window of 5 to 30 min after contrast administration, provided that the TI is adjusted. PMID- 16697322 TI - Racial differences in prevalence of coronary obstructions among men with positive nuclear imaging studies. AB - OBJECTIVES: The purpose of this research was to compare coronary obstruction between clinically similar African Americans (AA) and white persons undergoing coronary angiography. BACKGROUND: African Americans have higher rates of coronary death than whites, but are less likely to undergo coronary revascularization. Although differences in coronary anatomy do not explain racial difference in revascularization rates, several studies of clinically diverse persons undergoing coronary angiography have found less obstructive coronary disease in AA than clinically similar whites. METHODS: We studied 52 AA and 259 white male veterans who had both a positive nuclear perfusion imaging study and coronary angiography within 90 days of that study in five Department of Veterans Affairs hospitals. We used chart review and patient interview to collect demographics, clinical characteristics, and coronary anatomy results. Before angiography, we asked physicians to estimate the likelihood of coronary obstruction. RESULTS: The treating physicians' estimates of coronary disease likelihood were similar for AA (79.5%) and whites (83.0%); AA were less likely to have any coronary obstruction (63.5% vs. 76.5%, p = 0.05) and had significantly less severe coronary disease (p = 0.01) than whites. African Americans continued to be less likely to have coronary obstruction in analyses controlling for clinical features, including the physician's estimate of the likelihood of coronary obstruction. CONCLUSIONS: These results suggest that AA have less coronary obstruction than apparently clinically similar whites. Further studies are required to confirm our findings and better understand the paradox that AA are less likely to have obstructive coronary disease and more likely to suffer mortality from coronary disease. PMID- 16697323 TI - Assessment of left ventricular dyssynchrony in patients with conduction delay and idiopathic dilated cardiomyopathy: head-to-head comparison between tissue doppler imaging and velocity-encoded magnetic resonance imaging. AB - OBJECTIVES: This study sought to compare tissue Doppler imaging (TDI) with velocity-encoded (VE) magnetic resonance imaging (MRI) for left ventricular (LV) dyssynchrony assessment. BACKGROUND: Cardiac resynchronization therapy (CRT) is proposed for patients with heart failure, depressed LV function, and a wide QRS complex. Selection is based mainly on electrocardiogram criteria, but recent data suggest that intraventricular dyssynchrony may be preferred for selection. An LV dyssynchrony can adequately be assessed with TDI, but this has not been compared directly with other imaging modalities. A VE MRI potentially allows direct myocardial wall motion measurements similar to TDI. METHODS: Twenty patients with heart failure, systolic LV dysfunction, and a wide QRS complex were included, as well as 10 normal individuals with normal QRS duration and LV function. The TDI and VE MRI data were acquired to study intraventricular dyssynchrony. RESULTS: Left ventricular dyssynchrony was not observed in normal individuals (mean dyssynchrony -2 +/- 15 ms on TDI; mean -5 +/- 17 ms on MRI, p = NS). In patients, mean LV dyssynchrony was 55 +/- 37 ms on TDI; 49 +/- 38 ms on MRI (p = NS). Good correlation between both modalities was observed (linear regression TDI = 0.99 x MRI - 5, n = 30, r = 0.98, p < 0.01). The MRI showed a small, nonsignificant underestimation of 5 +/- 8 ms compared with TDI. Agreement between MRI and TDI for classification according to severity of LV dyssynchrony (minimal, intermediate, and extensive) was excellent (kappa +/- SE = 0.96 +/- 0.07, p < 0.01) with 95% of patients classified identical. CONCLUSIONS: Both MRI and TDI yield comparable information on LV dyssynchrony; MRI is useful in the selection of patients for CRT. PMID- 16697324 TI - Treatment of pulmonary arterial hypertension with the selective endothelin-A receptor antagonist sitaxsentan. AB - OBJECTIVES: We sought to determine the optimal dose of the selective endothelin A (ET(A)) receptor antagonist sitaxsentan for the treatment of pulmonary arterial hypertension (PAH); for observation only, an open-label (OL) bosentan arm was included. BACKGROUND: Endothelin is a mediator of PAH. In a preliminary PAH study, the selective ET(A) receptor antagonist sitaxsentan improved six-min walk (6MW) distance, World Health Organization (WHO) functional class (FC), and hemodynamics. METHODS: In this double-blind, placebo-controlled 18-week study, 247 PAH patients (idiopathic, or associated with connective tissue disease or congenital heart disease) were randomized; 245 patients were treated: placebo (n = 62), sitaxsentan 50 mg (n = 62) or 100 mg (n = 61), or OL (6MW tests, Borg dyspnea scores, and WHO FC assessments third-party blind) bosentan (n = 60). The primary end point was change in 6MW distance from baseline to week 18. Secondary end points included change in WHO FC, time to clinical worsening, and change in Borg dyspnea score. RESULTS: At week 18, patients treated with sitaxsentan 100 mg had an increased 6MW distance compared with the placebo group (31.4 m, p = 0.03), and an improved WHO FC (p = 0.04). The placebo-subtracted treatment effect for sitaxsentan 50 mg was 24.2 m (p = 0.07) and for OL bosentan, 29.5 m (p = 0.05). The incidence of elevated hepatic transaminases (>3x the upper limit of normal) was 6% for placebo, 5% for sitaxsentan 50 mg, 3% for sitaxsentan 100 mg, and 11% for bosentan. CONCLUSIONS: Treatment with the selective ET(A) receptor antagonist sitaxsentan, orally once daily at a dose of 100 mg, improves exercise capacity and WHO FC in PAH patients, with a low incidence of hepatic toxicity. PMID- 16697325 TI - Aortic valve regurgitation after arterial switch operation for transposition of the great arteries: incidence, risk factors, and outcome. AB - OBJECTIVES: The aims of this study were to assess the prevalence and incidence of aortic valve regurgitation (AR) after arterial switch operation (ASO), its outcome, and the risk factors. BACKGROUND: After an ASO, the long-term fate of the aortic valve is a concern as follow-up lengthens. METHODS: Operative and follow-up data on 1,156 hospital survivors after ASOs between 1982 and December 2000 were reviewed. RESULTS: At last follow-up (mean duration 76.2 +/- 60.5 months), 172 patients (14.9%) had an AR. Complex transposition of the great arteries, prior pulmonary banding done in 75 patients (21 with intact ventricular septum), aortic arch anomalies, AR at discharge, older age at ASO, and aortic/pulmonary size discrepancy were associated with AR. On multivariate analysis, the presence of a ventricular septal defect (VSD) or AR at discharge multiplied the risk by 2 and 4, respectively. Freedom from AR was 77.9% and 69.5% at 10 and 15 years, respectively; hazard function for AR declined rapidly and slowly increased thereafter. Reoperation from AR was done in 16 patients with one death, valvuloplasty being unsuccessful. Freedom from reoperation for AR was 97.7% and 96.8% at 10 and 15 years, respectively; hazard function slowly increased from 2 to 16 years. Higher late mortality was not associated with AR. CONCLUSIONS: After ASO, AR was observed and was related to VSD with attending high pressure and flow and AR at discharge. Progression of AR was slow, but incidence increased with follow-up. Reoperation for AR was rare. Late aortic valve function warrants long-term monitoring. PMID- 16697326 TI - Aortic regurgitation after arterial switch operation. PMID- 16697327 TI - Clinical outcome of 193 extracardiac Fontan patients: the first 15 years. AB - OBJECTIVES: We sought to evaluate the mid-term outcome of hospital survivors with extracardiac Fontan circulation. BACKGROUND: Few data exist about the mid-term and long-term results of the extracardiac Fontan operation. METHODS: From November 1988 to November 2003, 221 patients underwent an extracardiac Fontan procedure as primary (9 patients) or secondary (212 patients) palliation, at a mean age of 72.2 months (range 13.1 to 131.3 months). A total of 165 of 193 early survivors underwent programmed noninvasive follow-up evaluations and at least one cardiac catheterization. RESULTS: The overall survival, including operative deaths, was 85% at 15 years. Freedom from late failure among hospital survivors is 92% at 15 years. A total of 127 of 165 survivors (77%) were in New York Heart Association functional class I. The incidence of late major problems was 24% (42 major problems in 36 of 165 patients): 19 patients had arrhythmias (11%), 5 patients had obstruction of the extracardiac conduit (3%) and 6 of the left pulmonary artery (3.5%), and 5 patients experienced ventricular failure (3%), leading to heart transplantation in 3 patients. Protein-losing enteropathy was found in two patients (1%). The incidence of late re-interventions was 12.7% (21 of 165 patients, including 15 epicardial pacemaker implantations). Four patients died (2.3%), two after heart transplantation. CONCLUSIONS: After 15 years of follow-up, the overall survival, the functional status, and the cardiopulmonary performance of survivors of the extracardiac Fontan procedure compare favorably with other series of patients who underwent the lateral tunnel approach. The incidence of late deaths, obstructions of the cavopulmonary pathway, re interventions, and arrhythmias is lower than that reported late after other Fontan-type operations. PMID- 16697328 TI - Cellular basis for trigger and maintenance of ventricular fibrillation in the Brugada syndrome model: high-resolution optical mapping study. AB - OBJECTIVES: We examined how repolarization and depolarization abnormalities contribute to the development of extrasystoles and subsequent ventricular fibrillation (VF) in a model of the Brugada syndrome. BACKGROUND: Repolarization and depolarization abnormalities have been considered to be mechanisms of the coved-type ST-segment elevation (Brugada-electrocardiogram [ECG]) and development of VF in the Brugada syndrome. METHODS: We used high-resolution (256 x 256) optical mapping techniques to study arterially perfused canine right ventricular wedges (n = 20) in baseline and in the Brugada-ECG produced by administration of terfenadine (5 micromol/l), pinacidil (2 micromol/l), and pilsicainide (5 micromol/l). We recorded spontaneous episodes of phase 2 re-entrant (P2R) extrasystoles and subsequent self-terminating polymorphic ventricular tachycardia (PVT) or VF under the Brugada-ECG condition and analyzed the epicardial conduction velocity and action potential duration (APD) restitutions in each condition. RESULTS: Forty-one episodes of spontaneous P2R-extrasystoles in the Brugada-ECG were successfully mapped in 9 of 10 preparations, and 33 of them were originated from the maximum gradient of repolarization (GR(max): 176 +/- 54 ms/mm) area in the epicardium, leading to PVT (n = 12) or VF (n = 5). The epicardial GR(max) was not different between PVT and VF. Wave-break during the first P2R-extrasystole produced multiple wavelets in all VF cases, whereas no wave-break or wave-break followed by wave collision and termination occurred in PVT cases. Moreover, conduction velocity restitution was shifted lower and APD restitution was more variable in VF cases than in PVT cases. CONCLUSIONS: Steep repolarization gradient in the epicardium but not endocardium develops P2R extrasystoles in the Brugada-ECG condition, which might degenerate into VF by further depolarization and repolarization abnormalities. PMID- 16697329 TI - Autologous myoblast transplantation for chronic ischemic mitral regurgitation. AB - OBJECTIVES: This study was designed to assess whether post-myocardial infarction (MI) in-scar transplantation of skeletal myoblasts (SM) could reduce chronic ischemic mitral regurgitation (MR) by decreasing left ventricular (LV) remodeling. BACKGROUND: Extensive work has confirmed the relationship between ischemic MR and post-myocardial infarction (MI) remodeling of the LV. METHODS: An infero-posterior MI was created in 13 sheep, thereby resulting in increasing MR. Two months post-MI, the animals were randomized and in-scar injected with expanded autologous SM (n = 6, mean: 251 x 10(6) cells) or culture medium only (n = 7). Three-dimensional echocardiography was performed at baseline, before transplantation, and for two months thereafter (sacrifice), with measurements of LV end-diastolic and end-systolic volumes (ESV), ejection fraction (EF), MR stroke volume, and leaflet tethering distance; wall motion score index (WMSi) was assessed by two-dimensional echo. RESULTS: Measurements were similar between groups at baseline and before transplantation. At sacrifice, transplantation was found to have reduced MR progression (regurgitant volume change: -1.83 +/- 0.32 ml vs. 5.9 +/- 0.7 ml in control group, p < 0.0001) and tethering distance (-0.41 +/- 0.09 cm vs. 0.44 +/- 0.12 cm in control group, p < 0.001), with significant improvement of EF (2.01 +/- 0.94% vs. -4.86 +/- 2.23%, p = 0.02), WMSi (-0.25 +/- 0.11 vs. 0.13 +/- 0.03 in controls, p < 0.01) and a trend to a lesser increase in ESV (23.3 +/- 3.5 ml vs. 35.4 +/- 4.2 ml in control group, p = 0.055). CONCLUSIONS: Autologous skeletal myoblast transplantation attenuates mild-to moderate chronic ischemic MR, which otherwise is progressive, by decreasing tethering distance and improving EF and wall motion score, thereby enhancing valve coaptation. These data shed additional light on the mechanism by which skeletal myoblast transplantation may be cardioprotective. PMID- 16697330 TI - The year in cardiovascular surgery. PMID- 16697331 TI - Incomplete neointimal coverage of sirolimus-eluting stents: angioscopic findings. AB - OBJECTIVES: The goal of this study was to use angioscopy to investigate the amount of neointimal coverage after sirolimus-eluting stent (SES) implantation. BACKGROUND: Sirolimus-eluting stents reduce intimal hyperplasia. METHODS: We used angioscopy to evaluate 37 consecutive stented coronary artery lesions (15 SES and 22 bare-metal stents [BMS]) in 25 patients (18 men, 7 women) at 3 to 6 months after stent implantation. Angioscopic evaluation focused on: 1) neointimal coverage of stent struts, and 2) the existence of thrombi. The degree of neointimal coverage was classified as grade 0 when there was no neointimal coverage (similar to immediately after the implantation); grade 1 when stent struts bulged into the lumen, but were covered and still translucently visible; grade 2 when stent struts were visible but not clearly seen (not translucent); and grade 3 when stent struts were not visible because they were embedded in the neointima. RESULTS: Thrombi were identified in eight stented segments, tended to be more common with SES (p = 0.14), but were not seen on angiography. Three of the 15 SES (20%) had grade 0 neointimal coverage, and only 2 SES (13.3%) had complete coverage (grades 2/3). In contrast, all 22 BMS showed complete intimal coverage (grades 2/3). Thrombi were more common in stents with incomplete neointimal coverage (p = 0.09). CONCLUSIONS: The SES had incomplete neointimal coverage three to six months after implantation, and this was associated with subclinical thrombus formation. PMID- 16697332 TI - Drug-eluting stents and late adverse clinical outcomes lessons learned, lessons awaited. PMID- 16697333 TI - The perils of connectivity. PMID- 16697334 TI - The vascular smooth muscle cells apoptosis in asymptomatic diabetic carotid plaques: role of glycemic control. PMID- 16697335 TI - Evidence of cocaine-related coronary atherosclerosis in young patients with myocardial infarction. PMID- 16697336 TI - Association between right bundle branch block and impaired myocardial tissue level reperfusion in patients with acute myocardial infarction. PMID- 16697337 TI - Serum adiponectin levels are an independent predictor of the extent of coronary artery disease in men. PMID- 16697339 TI - Masked hypertension and white-coat hypertension prognosis. PMID- 16697340 TI - Training in cardiology fellowship programs. PMID- 16697342 TI - AHA/ACC guidelines for secondary prevention for patients with coronary and other atherosclerotic vascular disease: 2006 update endorsed by the National Heart, Lung, and Blood Institute. PMID- 16697343 TI - The molecular epidemiology of iridovirus in Murray cod (Maccullochella peelii peelii) and dwarf gourami (Colisa lalia) from distant biogeographical regions suggests a link between trade in ornamental fish and emerging iridoviral diseases. AB - Iridoviruses have emerged over 20 years to cause epizootics in finfish and amphibians in many countries. They may have originated in tropical Asia and spread through trade in farmed food fish or ornamental fish, but this has been difficult to prove. Consequently, MCP, ATPase and other viral genes were sequenced from archival formalin-fixed, paraffin-embedded tissues from farmed Murray cod (Maccullochella peelii peelii) that died during an epizootic in 2003 and from diseased gouramis that had been imported from Asia. There was almost complete homology (99.95%) over 4,527 bp between Murray cod iridovirus (MCIV) and an iridovirus (DGIV) present in dwarf gourami (Colisa lalia) that had died in aquarium shops in Australia in 2004, and very high homology with infectious spleen and kidney necrosis virus (ISKNV) (99.9%). These viruses are most likely to be a single species within the genus Megalocytivirus and probably have a common geographic origin. Primers for genus-specific PCR and for rapid discrimination of MCIV/DGIV/ISKNV and red sea bream iridovirus (RSIV), a notifiable pathogen, were developed. These were used in a survey to determine that the prevalence of DGIV infection in diseased gourami in retail aquarium shops in Sydney was 22% (95% confidence limits 15-31%). The global trade in ornamental fish may facilitate the spread of Megalocytivirus and enable emergence of disease in new host species in distant biogeographic regions. PMID- 16697344 TI - Assessing the effectiveness of "intuitive" vibrotactile warning signals in preventing front-to-rear-end collisions in a driving simulator. AB - This study was designed to investigate the possibility that driver responses to potential front-to-rear-end collision situations could be facilitated by implementing vibrotactile warning signals that indicate the likely direction of the potential collision. In a car following scenario in a driving simulator, participants drove along a rural road while trying to maintain a safe headway distance to the lead car using a visual distance display. Participants had to respond as quickly as possible to the sudden deceleration of the lead car which had its brake lights disabled, either with or without vibrotactile cues (presented in different experimental blocks). The results demonstrated significantly faster braking responses and larger safety margins when the vibrotactile warning signal was presented than when it was not. These findings demonstrate the effectiveness of vibrotactile cues in helping drivers to orient their spatial attention in the appropriate direction. Our results add to a growing body of empirical evidence highlighting the potential benefits of using "intuitive" vibrotactile in-car displays, in this case, to alert drivers to potential collisions and to provide time-critical directional information. PMID- 16697345 TI - Characterization of zero-valent iron nanoparticles. AB - The iron nanoparticle technology has received considerable attention for its potential applications in groundwater treatment and site remediation. Recent studies have demonstrated the efficacy of zero-valent iron nanoparticles for the transformation of halogenated organic contaminants and heavy metals. In this work, we present a systematic characterization of the iron nanoparticles prepared with the method of ferric iron reduction by sodium borohydride. Particle size, size distribution and surface composition were characterized by transmission electron microscopy (TEM), X-ray diffraction (XRD), high-resolution X-ray photoelectron spectroscopy (HR-XPS), X-ray absorption near edge structure (XANES) and acoustic/electroacoustic spectrometry. BET surface area, zeta (zeta) potential, iso-electric point (IEP), solution Eh and pH were also measured. Methods and results presented may foster better understanding, facilitate information exchange, and contribute to further research and development of iron nanoparticles for environmental and other applications. PMID- 16697346 TI - Comprehensive glycan analysis of recombinant Aspergillus niger endo polygalacturonase C. AB - The enzyme PGC is produced by the fungus Aspergillus niger during invasion of plant cell walls. The enzyme has been homologously overexpressed to provide sufficient quantities of purified enzyme for biological studies. We have characterized this enzyme in terms of its posttranslational modifications (PTMs) and found it to be both N- and O-glycosylated. The glycosyl moieties have also been characterized. This has involved a combination of matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF), liquid chromatography (LC)-ion trap, and LC-electrospray ionization (ESI) mass spectrometries in conjunction with trypsin degradation and beta-elimination, followed by Michael addition with dithiothreitol (BEMAD). This is the first demonstration of the ability of BEMAD to map glycosylation sites other than O-GlcNAc sites. The complete characterization of all PTMs on PGC allows us to model them on the peptide backbone, revealing potential roles played by the glycans in modulating the interaction of the enzyme with other macromolecules. PMID- 16697347 TI - Interaction of free fatty acids with mitochondria: coupling, uncoupling and permeability transition. AB - Long chain free fatty acids (FFA) exert, according to their actual concentration, different effects on the energy conserving system of mitochondria. Sub-micromolar concentrations of arachidonic acid (AA) rescue DeltapH-dependent depression of the proton pumping activity of the bc1 complex. This effect appears to be due to a direct interaction of AA with the proton-input mouth of the pump. At micromolar concentrations FFA increase the proton conductance of the inner membrane acting as protonophores. FFA can act as natural uncouplers, causing a mild uncoupling, which prevents reactive oxygen species production in the respiratory resting state. When Ca(2+)-loaded mitochondria are exposed to micromolar concentrations of FFA, the permeability of the inner membrane increases, resulting in matrix swelling, rupture of the outer membrane and release of intermembrane pro apoptotic proteins. The characteristics of AA-induced swelling appear markedly different in mitochondria isolated from heart or liver. While in the latter it presents the canonical features of the classical permeability transition (PT), in heart mitochondria substantial differences are observed concerning CsA sensitivity, DeltaPsi dependence, reversibility by BSA and specificity for the activating divalent cation. In heart mitochondria, the AA-dependent increase of the inner membrane permeability is affected by ANT ligands such as adenine nucleotides and atractyloside. AA apparently causes a Ca2+-mediated conversion of ANT from a translocator to a channel system. Upon diamide treatment of heart mitochondria, the Ca2+/AA-induced CsA insensitive channel is converted into the classical PT pore. The relevance of these observations in terms of tissue specific components of the putative PTP and heart ischemic and post-ischemic process is discussed. PMID- 16697348 TI - Mitochondrial DNA transcription and diseases: past, present and future. AB - The transcription of mitochondrial DNA has been studied for 30 years. However, many of the earlier observations are still unsolved. In this review we will recall the basis of mitochondrial DNA transcription, established more than twenty years ago, will include some of the recent progress in the understanding of this process and will suggest hypotheses for some of the unexplained topics. Moreover, we will show some examples of mitochondrial pathology due to altered transcription and RNA metabolism. PMID- 16697349 TI - Responsibility and obsessive-compulsive disorder: an experimental test. AB - To test the causal status of responsibility in obsessive-compulsive disorder (OCD), an experiment was executed in which responsibility was experimentally manipulated. OCD patients, non-OCD anxiety controls, and non-patients executed a classification task in either a high or a low responsibility (LoRes) condition. Subjective ratings related to danger and responsibility indicated that the manipulation was successful. Subjective OCD-like experiences and checking behaviors were higher in OCD patients in the high responsibility (HiRes) condition than in all other groups. Although the checking subscale of the Padua Inventory correlated with subjective ratings in the OCD patients in the HiRes condition, it was not associated with checking behaviors. The results confirm the hypothesis that responsibility plays a causal role in OCD. PMID- 16697350 TI - The relationship of momentary anger and impulsivity to bulimic behavior. AB - Past research has suggested that negative affect may be a causal factor for eating disordered behaviors. More specifically, research has shown that anger appears to be one aspect of negative affect that is particularly relevant in bulimic patients. Previous studies have also shown that the relationship between negative affect and eating disordered behaviors may partially depend upon personality variables such as impulsivity. The present study examined whether the relationship between anger and eating disordered behaviors is moderated by impulsivity. Subjects completed an ecological momentary assessment (EMA) protocol for an average of 2 weeks. Findings revealed that overall antecedent anger level and the variability of antecedent anger predicted binge-eating episodes and that these relationships were moderated by participants' level of impulsivity. These findings suggest that personality variables impact the way that anger and eating behaviors relate. They also suggest that the variability of antecedent anger may be a fruitful avenue for future research for those interested in causal variables associated with bulimia nervosa. PMID- 16697351 TI - Brain-derived neurotrophic factor conditional knockouts show gender differences in depression-related behaviors. AB - BACKGROUND: Indirect evidence suggests that loss of brain-derived neurotrophic factor (BDNF) from forebrain regions contributes to an individual's vulnerability for depression, whereas upregulation of BDNF in these regions is suggested to mediate the therapeutic effect of antidepressants. METHODS: We have tested this hypothesis by generating two independent lines of conditional BDNF knockout mice in which the BDNF gene is deleted selectively in forebrain. RESULTS: We show that male conditional knockouts exhibit hyperactivity but normal depression-related behaviors. In contrast, female conditional knockouts display normal locomotor activity but a striking increase in depression-like behavior. We also demonstrate that loss of BDNF in both male and female mice attenuates the actions of the antidepressant desipramine in the forced swim test. CONCLUSIONS: These gender differences in depression-related behaviors in BDNF conditional knockout mice provide direct evidence for a role of BDNF in depression. The results also support the view that forebrain BDNF may be essential in mediating antidepressant efficacy. PMID- 16697352 TI - Polymorphisms in the regulatory region of the human serotonin 5-HT2A receptor gene (HTR2A) influence gene expression. AB - BACKGROUND: Genomic variation in the regulatory region of the serotonin (5-HT) 2A receptor gene (HTR2A) may contribute to altered levels of 5-HT2A receptor and to psychiatric disease. METHODS: Frequency and linkage disequilibrium (LD) were determined for promoter single nucleotide polymorphisms (SNPs) -1438A/G, 1420C/T, and -783A/G in 156 subjects. Functional relevance of -1438A/G and 783A/G was assayed in vitro using a luciferase reporter assay and ex vivo using quantitative real time polymerase chain reaction in a set of human fibroblast cell lines. RESULTS: Significant LD was observed between SNPs -1438A/G and 783A/G. In vitro assays showed no significant differences in promoter activity between the A- and G-allele of -1438 locus when expressed with the major alleles at -1420C/T and -783A/G; however, when the minor allele G at -783 was expressed with G-allele at -1438, promoter activity was significantly decreased. 5-HT2A receptor mRNA expression in human fibroblast cell lines confirmed that -783A/G polymorphism significantly modified the effects of -1438A/G SNP. CONCLUSIONS: Our results demonstrate that SNP -783A/G modifies the effects of the major SNP 1438A/G. Future studies examining the association of -1438A/G polymorphism with diseases and 5-HT2A receptor expression analyses should account for this epistasis. PMID- 16697353 TI - Developmental vitamin D deficiency alters MK 801-induced hyperlocomotion in the adult rat: An animal model of schizophrenia. AB - BACKGROUND: Developmental vitamin D (DVD) deficiency has been proposed as a risk factor for schizophrenia. The behavioral phenotype of adult rats subjected to transient low prenatal vitamin D is characterized by spontaneous hyperlocomotion but normal prepulse inhibition of acoustic startle (PPI). The aim of this study was to examine the impact of selected psychotropic agents and one well-known antipsychotic agent on the behavioral phenotype of DVD deplete rats. METHODS: Control versus DVD deplete adult rats were assessed on holeboard, open field and PPI. In the open field, animals were given MK-801 and/or haloperidol. For PPI, the animals were given apomorphine or MK-801. RESULTS: DVD deplete rats had increased baseline locomotion on the holeboard task and increased locomotion in response to MK-801 compared to control rats. At low doses, haloperidol antagonized the MK-801 hyperactivity of DVD deplete rats preferentially and, at a high dose, resulted in a more pronounced reduction in spontaneous locomotion in DVD deplete rats. DVD depletion did not affect either baseline or drug-mediated PPI response. CONCLUSIONS: These results suggest that DVD deficiency is associated with a persistent alteration in neuronal systems associated with motor function but not those associated with sensory motor gating. In light of the putative association between low prenatal vitamin D and schizophrenia, the discrete behavioral differences associated with the DVD model may help elucidate the neurobiological correlates of schizophrenia. PMID- 16697354 TI - Long-term reduction in ventral tegmental area dopamine neuron population activity following repeated stimulant or ethanol treatment. AB - BACKGROUND: Drugs of abuse exert profound effects on the mesolimbic/mesocortical dopaminergic (DA) systems. Few studies have investigated the long-term adaptations in ventral tegmental area (VTA) DA neuron activity after repeated exposure to drugs of abuse. We investigated changes in the electrical activity of VTA DA neurons after cessation from treatment with several stimulants and ethanol. METHODS: Adult rats were treated with stimulants (amphetamine: 2 mg/kg per day, 5 days/week, 2 weeks; cocaine: 15 mg/kg per day, 5 days/week, 2 weeks; nicotine: .5 mg/kg per day, 5 days; ethanol: 10 g/kg per day, 3 weeks) and the single-unit activity of VTA DA neurons was studied in vivo 3 to 6 weeks later. RESULTS: Stimulant and ethanol treatment decreased basal VTA DA neuron population activity but not firing rate or firing pattern. This effect was reversed by acute apomorphine, suggesting that the underlying mechanism for reduced population activity was depolarization inactivation. Anesthesia did not confound this result, as similar effects were observed in amphetamine-treated rats recorded in a conscious preparation. CONCLUSIONS: Reduced basal VTA DA neuron population activity presumably due to depolarization inactivation is a common and long-term neuroadaptation to repeated treatment with stimulants and ethanol. This change in VTA DA neuron activity could underlie the persistent nature of addiction associated behaviors. PMID- 16697355 TI - Chronic treatment with mood stabilizers increases membrane GRK3 in rat frontal cortex. AB - BACKGROUND: G-protein receptor kinases (GRKs) are a family of serine/threonine kinases involved in the homologous desensitization of agonist activated G-protein coupled receptors (GPCRs). G-protein coupled receptor supersensitivity, possibly as a result of decreased GRK, has been suggested in affective disorders. METHODS: We used immunobloting to determine if chronic, therapeutically relevant doses of lithium (Li+), carbamazepine (CBZ), and valproate (VPA), would increase GRK2/3 protein levels in rat frontal cortex. RESULTS: Chronic Li+ (24%) and CBZ (44%) significantly increased GRK3 in the membrane but not cytosol fractions. Chronic VPA had no effect on GRK3. G-protein receptor kinase 2 protein levels were unchanged by all treatments. The GRK3 membrane to cytosol ratio was increased significantly in Li+ and CBZ treated rats. CONCLUSIONS: These results show that chronically administered Li+ and CBZ, but not VPA, increase the translocation of GRK3 from cytosol to membrane, possibly correcting supersensitivity of GPCRs in bipolar disorder. PMID- 16697356 TI - Sphere formation of ocular epithelial cells in the ciliary body is a reprogramming system for neural differentiation. AB - It is well known that neural stem/progenitor cells of the central nervous system (CNS) can proliferate to form neurospheres (CNS-neurospheres) that are positive for nestin, an intermediate filament for neural progenitors. Retinal stem/progenitor properties were also isolated from the ciliary body (CB) of the eye where, as in the CNS, such stem/progenitors also form spheres and have been considered to expand only via expansion by their proliferation even from the single-cell level (called spheres of pigment cells from the ciliary margin: PCM spheres). We here found a new and distinct process underlying the growth of CB cell-derived spheres (CB-spheres) that is unlike the mechanism of CNS- and PCM sphere expansion; this new process is a cell proliferation-independent incorporation of neighbor spheres and cells cultured at high density (200 cells/mul). The majority of cells in CB-spheres consisted of nestin-negative epithelia-like cells and started to express nestin during the course of their expansion by high-density cultivation. The growth of CNS-neurospheres was sensitive to a cell-cycle inhibitor, whereas the growth of CB-spheres was not seriously affected by cell proliferation; rather, the spheres grew by incorporating other CB-spheres and nestin-negative adherent cells, the latter of which started to express nestin and lost the expression of epithelial markers after being incorporated. These results indicate that CB-spheres do not form by the accumulation of neural progenitors but rather by a reprogramming system from epithelia-like cells for neural differentiation, a clearly distinct mechanism from sphere formation by single-cell expansion of retinal stem/progenitor populations. PMID- 16697357 TI - Differential expression of the CD14/TLR4 complex and inflammatory signaling molecules following i.c.v. administration of LPS. AB - The CD14/toll-like receptor 4 (TLR4) complex plays a vital role in initiating lipopolysaccharide (LPS) signaling during inflammation. In this study, we assessed innate immune responses and inflammatory transmission in the rat brain following intracerebroventricular (i.c.v.) administration of LPS. I.c.v. LPS induced the widespread increase in CD14 mRNA but did not change levels of TLR4 transcription in the brain. An increase in TLR4 immunoreactivity, coincident with cell death, leukocyte infiltration and neural tissue damage, was found in the meninges, choroid plexus and ventricular ependyma. In addition to CD14, rapid increases in gene expression of IkappaBalpha, IL-1beta, and TNF-alpha occurred along the meninges and ventricular ependyma. The response was most intense along the borders of the brain and declined in intensity in the adjacent periventricular areas and cerebral cortex. In the brain parenchyma, increased TLR4 immunoreactivity was confined to the vasculature and neighboring tissues along with strong vascular expression of IkappaBalpha and mPGES-1. These results suggest involvement of TLR4 in both brain inflammation and neural tissue injury and support the hypothesis that local diffusion and vascular transmission of inflammatory molecules are two major routes for developing inflammation in the brain. PMID- 16697358 TI - Heart muscle engineering: an update on cardiac muscle replacement therapy. AB - Cardiac muscle engineering aims at providing functional myocardium to repair diseased hearts and model cardiac development, physiology, and disease in vitro. Several enabling technologies have been established over the past 10 years to create functional myocardium. Although none of the presently employed technologies yields a perfect match of natural heart muscle, it can be anticipated that human heart muscle equivalents will become available after fine tuning of currently established tissue engineering concepts. This review provides an update on the state of cardiac muscle engineering and its utilization in cardiac regeneration. We discuss the application of stem cells including the allocation of autologous cell material, transgenic technologies that may improve tissue structure as well as in vivo engraftment, and vascularization concepts. We also touch on legal and economic aspects that have to be considered before engineered myocardium may eventually be applied in patients and discuss who may be a potential recipient. PMID- 16697359 TI - A novel ELISA system for selective measurement of human adiponectin multimers by using proteases. AB - BACKGROUND: Adiponectin, an antiatherogenic adipocyte-derived protein exists in human blood as multiple isoforms--trimeric low molecular weight (LMW), albumin binding LMW (Alb-LMW), hexameric middle molecular weight (MMW), and high molecular weight (HMW) forms. We developed a novel ELISA system to detect total human adiponectin and the selective level of each adiponectin multimer for investigating the distribution of these levels in human blood. METHODS: Two monoclonal antibodies that were raised against human adiponectin were used to construct a sandwich ELISA to measure adiponectin levels. Adiponectin multimers were selectively measured after sample pretreatment with two proteases that specifically digested the trimeric forms or both the hexameric and trimeric forms. RESULTS: The ELISA had a dynamic range of 0.075-4.8 ng/ml. Intraassay variations (CV) were 5.3% (total adiponectin), 4.1% (MMW+HMW), and 3.3% (HMW). Comparison of the results of ELISA and quantitative western blot analysis of multimeric adiponectin in serum samples revealed good correlation (LMW+Alb-LMW, r=0.873; MMW, r=0.907; HMW, r=0.950). Each of the three forms of adiponectin multimer levels closely correlated with total adiponectin levels in healthy subjects. CONCLUSIONS: This ELISA system can be used to further investigate the physiological roles of human adiponectin multimers. PMID- 16697360 TI - New "plasma referenced" POCT glucose monitoring systems--are they suitable for glucose monitoring and diagnosis of diabetes? AB - BACKGROUND: Results from portable glucose meters should be reported as plasma values as recommended by IFCC. Three new "plasma calibrated" blood glucose meters (Abbott Precision Xcceed, Bayer Ascensia Contour and Roche Accu-Chek Aviva) were compared against laboratory venous plasma glucose measurements to determine their suitability for patient monitoring and diabetes diagnosis. METHODS: 115 capillary blood samples were analyzed on each of the three meters and compared to venous plasma measurements on the Dade Behring RXL analyzer (Hexokinase method). RESULTS: We found a significant positive bias of all meters compared to the laboratory reference method. Deviations of more than 10% were seen in more than a third of all glucose values in all three devices. 13%, 8.7% and 10.4%, respectively, of the values from the Abbott, Bayer and Roche devices fell in zone B or C of the Parkes' error grid. CONCLUSIONS: Neither glucose meter met the ADA or the NACB goals and should therefore not be used as screening tests to diagnose diabetes but are well suitable for patient monitoring. PMID- 16697361 TI - Clinical laboratory findings associated with in-hospital mortality. AB - The diagnostic approach and the clinical management of critically ill patients is challenging. The recognition of biomarkers related to in-hospital mortality is of importance for identification of patients at increased risk of death. Many prediction models assessing the severity of illness and likelihood of hospital survival were developed using logistic regression analyses. These models include several laboratory parameters, such as white blood cell counts, serum bilirubin, serum albumin, blood glucose, serum electrolytes and markers which reflect acid base disturbances. Recently, several other biomarkers, including troponin, B-type natriuretic peptide (BNP), N-terminal proBNP, C-reactive protein, procalcitonin, cholesterol and coagulation related markers have emerged as clinically useful tools for risk stratification and mortality prediction of heterogeneous and more specific subgroups of critically ill patients. More investigations are required to verify whether risk stratification based on mortality-related biomarkers may translate into targeted treatment strategies to improve clinical outcome of the critical illness. Biomarkers which are related to in-hospital mortality are highlighted in the current review. PMID- 16697362 TI - Biological and clinical aspects of the vitamin D binding protein (Gc-globulin) and its polymorphism. AB - The vitamin D binding protein (DBP) is the major plasma carrier protein of vitamin D and its metabolites. Unlike other hydrophobic hormone-binding systems, it circulates in a considerably higher titer compared to its ligands. Apart from its specific sterol binding capacity, DBP exerts several other important biological functions such as actin scavenging, fatty acid transport, macrophage activation and chemotaxis. The DBP-gene is a member of a multigene cluster that includes albumin, alpha-fetoprotein, and alpha-albumin/afamin. All four genes are expressed predominantly in the liver with overlapping developmental profiles. DBP is a highly polymorphic serum protein with three common alleles (Gc1F, Gc1S and Gc2) and more than 120 rare variants. The presence of unique alleles is a useful tool for anthropological studies to discriminate and to reveal ancestral links between populations. Many studies have discussed the link between DBP-phenotypes and susceptibility or resistance to osteoporosis, Graves' disease, Hashimoto's thyroiditis, diabetes, COPD, AIDS, multiple sclerosis, sarcoidosis and rheumatic fever. This article reviews the general characteristics, functions and clinical aspects of DBP. PMID- 16697363 TI - An improved matched filter for blood vessel detection of digital retinal images. AB - The matched filter has been widely used in the detection of blood vessels of the human retina digital image. In this paper, the matched filter response to the detection of blood vessels is increased by proposing better filter parameters. These filter parameters are found by using an optimization procedure on 20 retina images of the DRIVE database. Comparisons with other approaches show that the matched filter that uses the newly found parameters outperforms the matched filter that uses the classical filter parameters as well as some vessel detection techniques. A technique is also discussed to find the best threshold value for the continuous matched filter output image and hence the best segmented vessel image. PMID- 16697364 TI - Developmental expression and hormonal regulation of different isoforms of the transcription factor E75 in the tobacco hornworm Manduca sexta. AB - E75A and E75B, isoforms of the E75 orphan nuclear receptor, are sequentially up regulated in the abdominal epidermis of the tobacco hornworm Manduca sexta by 20 hydroxyecdysone (20E) during larval and pupal molts, with E75A also increasing at pupal commitment (Zhou et al., Dev. Biol. 193, 127-138, 1998). We have now cloned E75C and show that little is expressed in the epidermis during larval life with trace amounts seen just before ecdysis. Instead, E75C is found in high amounts during the development of the adult wings as the ecdysteroid titer is rising, and this increase was prevented by juvenile hormone (JH) that prevented adult development. By contrast, E75D is expressed transiently during the larval and pupal molts as the ecdysteroid titer begins to decline and again just before ecdysis, but in the developing adult wings is expressed on the rise of 20E. Removal of the source of JH had little effect on either E75C or E75D mRNA expression during the larval and pupal molts. At the time of pupal commitment, in vitro experiments show that 20E up-regulates E75D and JH prevents this increase. Neither E75A nor E75D mRNA was up-regulated by JH alone. Thus, E75C is primarily involved in adult differentiation whereas E75D has roles both during the molt and pupal commitment. PMID- 16697365 TI - Salusin beta is a surrogate ligand of the mas-like G protein-coupled receptor MrgA1. AB - The mas-like G protein-coupled receptors form a subfamily of G protein-coupled receptors that includes variable member numbers across different species and that have been shown to bind a wide variety of ligands from peptides to amino acid derivatives. While screening a library of peptides against different orphan G protein-coupled receptors, we found that human salusin beta activates the mouse mas-like G protein-coupled receptor, mMrgA1 with an EC(50) of about 300 nM. Salusin beta is a bioactive peptide recently discovered through bioinformatics analysis which stimulates arginine-vasopressin release from rat pituitary and causes rapid and profound hypotension and bradycardia. However, when we further analyzed the generality of the mMrgA1 activation, we found that human salusin beta does not activate corresponding human mas-like G protein-coupled receptors. Our results show that human salusin beta is a surrogate ligand of the mouse MrgA1 and raises a cautionary flag for experiments that analyze the pharmacological profiles of mas-like G protein-coupled receptors from different species. PMID- 16697366 TI - Analgesic effects of the somatostatin sst4 receptor selective agonist J-2156 in acute and chronic pain models. AB - Somatostatin released from capsaicin-sensitive afferents exerts systemic anti nociceptive actions, presumably via somatostatin receptor subtype 4 (sst4). In the present study, the antinociceptive effects of a novel somatostatin sst4 receptor selective peptidomimetic compound, J-2156 (1-100 microg/kg i.p.), were examined. J-2156 inhibited nocifensive behaviour of mice in the second phase of the formalin test. Adjuvant-evoked chronic inflammatory mechanical allodynia was decreased in rats treated with J-2156 for 21 days. Sciatic nerve ligation-induced neuropathic mechanical hyperalgesia was inhibited by J-2156 on the seventh postoperative day. Results obtained using this highly selective agonist suggest that somatostatin sst4 receptors represent a promising target for new perspectives in analgesic therapy. PMID- 16697367 TI - Signal peptide peptidase dependent cleavage of type II transmembrane substrates releases intracellular and extracellular signals. AB - The intramembrane-cleaving proteases (I-CLiPs) presenilin-1 and -2 (PS1 and PS2), signal peptide peptidase (SPP) and the Site-2 protease (S2P) catalyze critical steps in cell signaling and are implicated in diseases such as Alzheimer's disease, hepatitis C virus (HCV) infection and cholesterol homeostasis. Here we describe the development of a cellular assay based on cleavage of the transmembrane sequence of the HCV core protein precursor, releasing intra- and extra-cellular signals that represent sequential signal peptidase and SPP cleavage, respectively. We find that the SPP inhibitor (Z-LL)2-ketone (IC50 = 1.33 microM) and the gamma-secretase potent inhibitors NVP-AHW700-NX (IC50 = 51 nM) and LY411575 (IC50 = 61 nM) but not DAPT dose dependently inhibited SPP but not signal peptidase cleavage. Our data confirm that type II orientated substrates, like the HCV transmembrane sequence, are sequentially cleaved by signal peptidase then SPP. This dual assay provides a powerful tool to pharmacologically analyze sequential cleavage events of signal peptidase and SPP and their regulation. PMID- 16697368 TI - Focus on molecules: rod photoreceptor cGMP-gated cation channel. PMID- 16697369 TI - High glucose concentration induces elevated expression of anti-oxidant and proteolytic enzymes in cultured human retinal pigment epithelial cells. AB - We investigated the differential protein expression patterns of retinal pigment epithelial (RPE) cells exposed to increased glucose concentrations. Cultured human RPE cells (ARPE-19) were exposed for 4 days with normal blood glucose concentration (5.5 mM D-glucose), followed by exposure to either normal (5.5 mM) or high (33 mM) concentrations of D-glucose for 48h. Protein extracts of glucose treated RPE cells were then subjected to comparative proteome analysis based on 2 D gel electrophoresis. Protein spots were visualized by silver staining. The differentially expressed proteins were excised and digested in-gel with trypsin, then analysed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The expression levels of cathepsin B, glutathione peroxidase and heat shock protein 27 were increased, and that of protein disulfide isomerase decreased in high glucose treated RPE compared to normal glucose. The isoelectric point of copper/zinc-containing superoxide dismutase (Cu/Zn-SOD) shifted toward acidic region in response to high glucose. Cu/Zn-SOD activity in high glucose group was significantly lower than that in normal glucose group (P<0.05, Mann-Whitney U-test). Systematic survey of protein expression has revealed that RPE cells respond to acute, pathologically high glucose levels by the elevated expression of anti-oxidant and proteolytic enzymes. PMID- 16697370 TI - Glial activation in white matter following ischemia in the neonatal P7 rat brain. AB - This study examines cell death and proliferation in the white matter after neonatal stroke. In postnatal day 7 injured rat, there was a marked reduction in myelin basic protein (MBP) immunostaining mainly corresponding to numerous pyknotic immature oligodendrocytes and TUNEL-positive astrocytes in the ipsilateral external capsule. In contrast, a substantial restoration of MBP, as indicated by the MBP ratio of left-to-right, occurred in the cingulum at 48 (1.27 +/- 0.12) and 72 (1.30 +/- 0.18, P < 0.05) h of recovery as compared to age matched controls (1.03 +/- 0.14). Ki-67 immunostaining revealed a first peak of newly generated cells in the dorsolateral hippocampal subventricular zone and cingulum at 72 h after reperfusion. Double immunofluorescence revealed that most of the Ki-67-positive cells were astrocytes at 48 h and NG2 pre-oligodendrocytes at 72 h of recovery. Microglia infiltration occurs over several days in the cingulum, and a huge quantity of macrophages reached the subcortical white matter where they engulfed immature oligodendrocytes. The overall results suggest that the persistent activation of microglia involves a chronic component of immunoinflammation, which overwhelms repair processes and contributes to cystic growth in the developing brain. PMID- 16697371 TI - Mechanisms of homocysteine neurotoxicity in neurodegenerative diseases with special reference to dementia. AB - Mild to moderate hyperhomocysteinemia is a risk factor for neurodegenerative diseases. Human studies suggest that homocysteine (Hcy) plays a role in brain damage, cognitive and memory decline. Numerous studies in recent years investigated the role of Hcy as a cause of brain damage. Hcy itself or folate and vitamin B12 deficiency can cause disturbed methylation and/or redox potentials, thus promoting calcium influx, amyloid and tau protein accumulation, apoptosis, and neuronal death. The Hcy effect may also be mediated by activating the N methyl-D-aspartate receptor subtype. Numerous neurotoxic effects of Hcy can be blocked by folate, glutamate receptor antagonists, or various antioxidants. This review describes the most important mechanisms of Hcy neurotoxicity and pharmacological agents known to reverse Hcy effects. PMID- 16697372 TI - Membrane leakage induced by dynorphins. AB - Dynorphins, endogeneous opioid peptides, function as ligands to the opioid kappa receptors and induce non-opioid excitotoxic effects. Here we show that big dynorphin and dynorphin A, but not dynorphin B, cause leakage effects in large unilamellar phospholipid vesicles (LUVs). The effects parallel the previously studied potency of dynorphins to translocate through biological membranes. Calcein leakage caused by dynorphin A from LUVs with varying POPG/POPC molar ratios was promoted by higher phospholipid headgroup charges, suggesting that electrostatic interactions are important for the effects. A possibility that dynorphins generate non-opioid excitatory effects by inducing perturbations in the lipid bilayer of the plasma membrane is discussed. PMID- 16697373 TI - Interaction of nucleoside diphosphate kinase and catalases for stress and light responses in Neurospora crassa. AB - Nucleoside diphosphate kinase (NDK) is an ubiquitous enzyme with the function of a signal transducer. In Neurospora crassa, an ndk-1(P72H) mutant carrying the point mutation Pro72His was isolated. We found that ndk-1(P72H) showed hypersensitivity to oxidative and heat stress and a decrease in the levels of catalase (Cat)-1 and -3 induced by oxidative, heat stress and illumination compared with wild type (Wt). We found, by conducting a yeast two-hybrid assay, that Cat-1 interacted with NDK-1. NDK-1 was suggested to control Cat-1 and Cat-3 at the post-transcriptional level in response to heat, oxidative stress and light. PMID- 16697374 TI - Short-term retention of actin filament binding proteins on lamellipodial actin bundles. AB - Actin filaments are organised into sub-compartments of meshwork and bundles in lamellipodia. Localisation of fascin, the LIM and SH3 domain protein 1 (lasp-1), and lasp-2 to the bundles suggest their involvement in that organisation; however, their contributions remain unclear. We have compared the turnover of these proteins with actin at the bundle. After photobleaching, EGFP-actin recovered inwards from the bundle tip, consistent with the retrograde flow by treadmilling. In contrast, the recovery of EGFP-fascin, -lasp-1 and -lasp-2 occurred from the anterograde direction. These results suggest that these molecules would participate in the stabilisation of bundles but not in initiation. PMID- 16697375 TI - Identification of TNF-alpha-responsive NF-kappaB p65-binding element in the distal promoter of the mouse serine protease inhibitor SerpinE2. AB - Serine protease inhibitor SerpinE2 is known as a cytokine-inducible gene. Here, we investigated whether tumor necrosis factor alpha-(TNF-alpha)-induced expression of SerpinE2 is mediated by the nuclear factor-kappaB (NF-kappaB) p65 subunit. Both steady state and TNF-alpha-induced expression of SerpinE2 mRNA were abrogated in p65-/- murine embryonic fibroblasts (MEFs). Reconstitution with wild type p65 rescued SerpinE2 mRNA expression in an IkappaB kinase beta-dependent manner. Electrophoresis mobility shift assay and ChIP assay demonstrated that p65 bound to the kappaB-like DNA sequence located at approximately -9 kbp in the SerpinE2 promoter. In addition, TNF-alpha stimulated luciferase gene expression driven by the kappaB-like element in the reconstituted MEFs, but not in p65-/- MEFs. These results indicated that activation of NF-kappaB p65 plays an important role in TNF-alpha-induced expression of SerpinE2. PMID- 16697376 TI - Functional dissection of human protease mu-calpain in cell migration using RNAi. AB - Calpains are a family of calcium-dependent cysteine proteases involved in a variety of cellular functions. Two isoforms, m-calpain and mu-calpain, have been implicated in cell migration. However, since conventional inhibitors used for the studies of the functions of these enzymes lack specificity, the individual physiological function and biochemical mechanism of these two isoforms, especially mu-calpain, are not clear. In contrast, RNA interference has the potential to allow a sequence-specific destruction of target RNA for functional assay of gene of interest. In the present study, we found that small interfering RNAs-mediated knockdown of mu-calpain expression in MCF-7 cells that do not express m-Calpain led to a reduction of cell migration. This isoform-specific function of mu-calpain was further confirmed by the rescue experiment as overexpression of mu-calpain but not m-calpain could restore the cell migration rate. Knockdown of mu-calpain also altered cell morphology with increased filopodial projections and a highly elongated tail that seemed to prevent cell spreading and migration with reduced rear detachment ability. Furthermore, knockdown of mu-calpain decreased the proteolytic products of filamin and talin, which were specifically rescued by overexpression of mu-calpain but not m calpain, suggesting that their proteolysis could be one of the key mechanisms by which mu-calpain regulates cell migration. PMID- 16697377 TI - Proteomic analysis of UVC irradiation-induced damage of plasma proteins: Serum amyloid P component as a major target of photolysis. AB - Ultraviolet-C (UVC) irradiation is a pathogen inactivation method used for disinfection of pharmaceutical products derived from human blood. Previous studies have shown that UVC can potentially damage proteins through photolysis or can generate reactive species resulting in protein thiol oxidation. In this study, two fluorescence-based quantitative proteomic approaches were used to assess the effects of a novel UVC-disinfection strategy on human plasma fractions. We show minimal changes in protein content, but gross alterations in protein thiol reactivity, indicative of oxidative damage. We identify a number of the damaged proteins by mass spectrometry, including serum amyloid P component, and further demonstrate UVC-induced photolysis of its disulphide bond. PMID- 16697378 TI - A transgenic rice seed accumulating an anti-hypertensive peptide reduces the blood pressure of spontaneously hypertensive rats. AB - RPLKPW is a potent anti-hypertensive peptide designed according to the structure of ovokinin(2-7) (RADHPF). In this study, we generated transgenic rice plants that accumulate the RPLKPW peptide as a fusion protein with the rice storage protein glutelin. The engineered peptide is expressed under the control of endosperm-specific glutelin promoters and specifically accumulates in seeds. Oral administration of either the RPLKPW-glutelin fraction or transgenic rice seeds to spontaneously hypertensive rats (SHRs) significantly reduced systolic blood pressures. These results suggest the possible application of transgenic rice seed as a nutraceutical delivery system and specifically for administration of active peptides in hypertension. PMID- 16697379 TI - Comparative analysis of enzyme activities and mRNA levels of peptidyl prolyl cis/trans isomerases in various organs of wild type and Pin1-/- mice. AB - We investigated the enzyme activity of peptidyl prolyl cis/trans isomerases (PPIases) in brain, testis, lung, liver, and mouse embryonic fibroblasts (MEF) of Pin1+/+ and Pin1-/- mice. The aim of this study is to determine if other PPIases can substitute for the loss of Pin1 activity in Pin1-/- mice and what influence Pin1 depletion has on the activities of other PPIases members. The results show that high PPIase activities of Pin1 are found in organs that have the tendency to develop Pin1 knockout phenotypes and, therefore, provide for the first time an enzymological basis for these observations. Furthermore we determined the specific activity (k(cat)/K(M)) of endogenous Pin1 and found that it is strongly reduced as compared with the recombinant protein in all investigated organs. These results suggest that posttranslational modifications may influence the PPIase activity in vivo. The activities originating from cyclophilin and FKBP are not influenced by the Pin1 knockout, but a basal enzymatic activity towards phosphorylated substrates could be found in Pin1-/- lysates. Real time PCR experiments of all PPIases in different mouse organs and MEF of Pin1+/+ and Pin1 /- mice support the finding and reveal the specific expression profiles of PPIases in mice. PMID- 16697380 TI - Hsp70 inhibits lipopolysaccharide-induced NF-kappaB activation by interacting with TRAF6 and inhibiting its ubiquitination. AB - Inducible heat shock protein 70 (Hsp70) is one of the most important HSPs for maintenance of cell integrity during normal cellular growth as well as pathophysiological conditions. Tumor necrosis factor receptor-associated factor 6 (TRAF6) is a crucial signaling transducer that regulates a diverse array of physiological and pathological processes and is essential for activating NF kappaB signaling pathway in response to bacterial lipopolysaccharide (LPS). Here we report a novel mechanism of Hsp70 for preventing LPS-induced NF-kappaB activation in RAW264.7 macrophage-like cells. Our results show that Hsp70 can associate with TRAF6 physically in the TRAF-C domain and prevent TRAF6 ubiquitination. The stimulation of LPS dissociates the binding of Hsp70 and TRAF6 in a time-dependent manner. Hsp70 inhibits LPS-induced NF-kappaB signaling cascade activation in heat-shock treated as well as Hsp70 stable transfected RAW264.7 cells and subsequently decreases iNOS and COX-2 expression. Two Hsp70 mutants, Hsp70DeltaC(1-428aa) with N-terminal ATPase domain and Hsp70C(428-642aa) with C-terminal domain, lack the ability to influence TRAF6 ubiquitination and TRAF6-triggered NF-kappaB activation. Taken together, these findings indicate that Hsp70 inhibits LPS-induced NF-kappaB activation by binding TRAF6 and preventing its ubiquitination, and results in inhibition of inflammatory mediator production, which provides a new insight for analyzing the effects of Hsp70 on LPS-triggered inflammatory signal transduction pathways. PMID- 16697381 TI - Individual variation in cortisol responses to acute "on-back" restraint in an outbred hamster. AB - An outbred species of dwarf hamster (Phodopus campbelli) was used to assess between-individual variability in the response to, and recovery from, a one-time stressor of 6 min of physical restraint in a subordinate, on-back, position. Four repeated plasma samples were drawn under home-cage isoflurane anesthesia from 33 males and 38 females 50 min before, and then 10, 60, and 120 min after the stress onset. Plasma cortisol concentrations were higher in females than males, but there was no evidence for a sex difference in response to the stressor. The expected cross-sectional increase ( approximately 50 ng/ml) in response to the stressor, followed by recovery, was seen. However, there was extensive individual variation, ranging from no reaction to continuous decline from the initial to the final sample. Results were expressed in four ways (absolute concentration, relative concentration, and area under the curve relative to ground and relative to the stress-induced increase) and also standardized and subjected to hierarchical cluster analysis. Clusters failed to effectively partition the between-individual variation and did not cluster by sex, age, or housing conditions. The current study cautions against ignoring individual differences and suggests that outbred animal models might be particularly relevant to understanding stress-related pathological conditions. PMID- 16697382 TI - Angiotensin II disrupts inhibitory avoidance memory retrieval. AB - The brain renin-angiotensin system (RAS) is involved in learning and memory, but the actual role of angiotensin II (A(II)) and its metabolites in this process has been difficult to comprehend. This has been so mainly due to procedural issues, especially the use of multi-trial learning paradigms and the utilization of pre training intracerebroventricular infusion of RAS-acting compounds. Here, we specifically analyzed the action of A(II) in aversive memory retrieval using a hippocampal-dependent, one-trial, step-down inhibitory avoidance task (IA) in combination with stereotaxically localized intrahippocampal infusion of drugs. Rats bilaterally implanted with infusion cannulae aimed to the CA1 region of the dorsal hippocampus were trained in IA and tested for memory retention 24 h later. We found that when given into CA1 15 min before IA memory retention test, A(II), but not angiotensin IV or angiotensin(1-7) induced a dose-dependent and reversible amnesia without altering locomotor activity, exploratory behavior or anxiety state. The effect of A(II) was blocked in a dose-dependent manner by the A(II)-type 2 receptor (AT(2)) antagonist PD123319 but not by the A(II)-type 1 receptor (AT(1)) antagonist losartan. By themselves, neither PD123319 nor losartan had any effect on memory expression. Our data indicate that intra-CA1 A(II) hinders retrieval of avoidance memory through a process that involves activation of AT(2) receptors. PMID- 16697383 TI - Psychosocial outcomes of Hong Kong Chinese diagnosed with acute coronary syndromes: a prospective repeated measures study. AB - BACKGROUND: Western studies have suggested that emotional stress and distress impacted on the morbidity and mortality in people following acute coronary events. Symptoms of anxiety and depression have been associated with re infarction and death, prolonged recovery and disability and depression may precipitate the client's low self-esteem. This study examined perceived anxiety, depression and self-esteem of Hong Kong Chinese clients diagnosed with acute coronary syndrome (ACS) over a 6-month period following hospital admission. OBJECTIVES: To examine: DESIGN: A prospective, repeated measures design with measures taken on two occasions over a 6-month period; (1) within the 1st week of hospital admission following the onset of ACS and (2) at 6 months follow up. SETTING AND PARTICIPANTS: Convenient sample of 182 voluntary consented clients admitted with ACS to a major public hospital in Hong Kong who could communicate in Chinese, complete questionnaires, cognitive intact, and were haemodynamically stable and free from acute chest pain at the time of interview. METHODS: Baseline data were obtained within 1 week after hospital admission. The follow-up data was collected 6 months after hospital discharge. The Chinese version of the Hospital Anxiety and Depression Scale (HADS), State Self-esteem Scale (SSES), and Rosenberg's Self-Esteem Scale (RSES) were used to assess anxiety and depression, state self-esteem, and trait self-esteem, respectively. RESULTS: Findings suggested gender differences in clients' perception in anxiety, depression and self-esteem. Improvements in clients' perception of these variables were evident over the 6-month period following their acute coronary events. CONCLUSION: The study confirmed the western notion that psychosocial problems are common among coronary clients and this also applies to Hong Kong Chinese diagnosed with ACS. Further studies to explore effective interventions to address these psychosocial issues are recommended. PMID- 16697385 TI - Induction of tissue factor expression and release as microparticles in ECV304 cell line by Chlamydia pneumoniae infection. AB - The association between Chlamydia pneumoniae (C. pneumoniae) infection and the onset and progression of atherosclerosis has become apparent recently. Moreover, increased expression of tissue factor (TF) as a result of C. pneumoniae infection has been previously demonstrated. We have examined the expression of TF on the surface of endothelial cells and the release of TF-containing cell-derived microparticles, over seven days. Additionally, using cells expressing a procoagulantly active EGFP-TF hybrid protein, we examined the kinetics of TF trafficking on the cells and incorporation into shed microparticles. Finally, in an attempt to associate this with the activation of NFkappaB, we used a luciferase reporter to measure the duration of the activation of this transcription factor. TF-containing microparticles were released within 24h of infection and continued for up to 7 days. Moreover, the initial release of TF containing microparticles was associated with NFkappaB activation and was suppressed on inclusion of an NFkappaB inhibitor, pyrrolidinedithiocarbamate ammonium. Moreover, persistent dissemination of TF-containing microparticles at later stages of infection was associated with the release of the infective C. pneumoniae elementary bodies. The released procoagulant, cellular microparticles are known to be strongly atherogenic and therefore we suggest a mechanism for the involvement of C. pneumoniae in the onset and progression of vascular disease. PMID- 16697386 TI - Analysis of the effect of multiple genetic variants of cardiovascular disease risk on insulin concentration variability in healthy adults of the STANISLAS cohort. The role of FGB-455 G/A polymorphism. AB - INTRODUCTION: Given the hypothesis of a common soil for atherosclerosis, type 2 diabetes and metabolic syndrome, we tested the contribution of gene polymorphisms involved in cardiovascular diseases on fasting insulin concentration (FIC). METHODS: The polymorphisms were investigated by a multiplex assay in 308 apparently healthy French middle-aged men and women, taken from the STANISLAS cohort. FIC was measured by a microparticular enzymatic immunoassay. RESULTS: After a series of regression analyses involving 34 polymorphisms, FGB -455G/A was the only polymorphism that remained significantly associated with FIC when adjusting the analyses for multiple testing. Stepwise models showed that FGB polymorphism accounted for 4.39% of FIC variability in men. Additionally, interactions between FGB and with environmental factors (alcohol and smoking in men, and BMI in women) were found. DISCUSSION: To our knowledge, this is the first study reporting an influence of FGB polymorphism on FIC in a healthy population. Our results concord with the already shown link between fibrinogen concentration and FIC, and support the hypothesis of a relationship between fibrinogen and endothelium in FIC homeostasis whose alteration may induce several metabolic disorders. The contribution of this gene, although modest, is consistent with the polygenic nature of insulin levels. PMID- 16697387 TI - Posterior and anterior components of force during bite loading. AB - Late anterior crowding of teeth has been associated with the anterior component of force (ACF) developed during biting. Possible physiologic mechanisms countering ACF, including the presence of a posterior component of force (PCF), are hypothesized. In this self-controlled study, 60 subjects aged 27.05+/-3.9 years were examined for ACF and PCF that were calculated as the change in tightness of a mandibular dental contact points from non-biting to biting state. Both ACF and PCF were found to develop simultaneously. However, the PCF was 4-7 folds smaller than the ACF (p<0.001). The ACF progressively declined by 10-20 folds (p<0.001) from the posterior to anterior dentition. The lateral incisor canine contact point had the greatest ACF decline (63-74%). ACF effect on the anterior dentition is counteracted by a protective mechanism consisted of PCF, progressive dissipation of ACF, and canine blockage. PMID- 16697388 TI - Simple and rapid determination of anatoxin-a in lake water and fish muscle tissue by liquid-chromatography-tandem mass spectrometry. AB - Anatoxin-a (AN) is a powerful neurotoxin that can be produced by cyanobacteria in eutrophic freshwaters. Consequently, AN can contaminate lakes, rivers and basins destined for drinking water and aquaculture. Two simple, specific and sensitive procedures for determining AN in lake water and fish muscle tissue are presented. Both analytical protocols are based on liquid-chromatography (LC)-tandem mass spectrometry (MS) with electrospray ionization. MS data were acquired in the multi reaction monitoring mode by selecting four precursor to product ion transitions. After filtration, AN in lake water was analyzed by directly injecting 0.5 ml of the aqueous sample in the LC column. Analysis of AN in fish muscle tissue involved the matrix solid-phase dispersion technique. The analyte was extracted from tissue by 4 ml of water acidified to pH 2 and heated at 80 degrees C. After acidification and filtration, 0.2 ml of the aqueous extract was injected in the LC column. Analyte recovery ranged between 71 and 79% and was not substantially affected by both the analyte concentration and the type of fish. Phenylalanine is an essential amino acid invariably present in any animal tissue. Like AN, this amino acid produces a pseudo molecular ion at m/z 166, it has a very similar fragmentation pattern and LC retention time. This method is able to prevent identifying phenylalanine for AN as the latter compound is eluted more than 1 min before the former one and the two compounds have remarkably different relative ion signal intensities. On the basis of a signal-to-noise ratio of 10, limits of quantification of AN in water and fish fillet were estimated to be 13 ng/l and 0.5 ng/g, respectively. PMID- 16697384 TI - Identification and analysis of genes expressed in the adult filarial parasitic nematode Dirofilaria immitis. AB - The heartworm Dirofilaria immitis is a filarial parasitic nematode infecting dogs and other mammals worldwide causing fatal complications. Here, we present the first large-scale survey of the adult heartworm transcriptome by generation and analysis of 4005 expressed sequence tags, identifying about 1800 genes and expanding the available sequence information for the parasite significantly. Brugia malayi genomic data offered the most valuable information to interpret heartworm genes, with about 70% of D. immitis genes showing significant similarities to the assembly. Comparative genomic analyses revealed both genes common to metazoans or nematodes and genes specific to filarial parasites that may relate to parasitism. Characterization of abundant transcripts suggested important roles for genes involved in energy generation and antioxidant defense in adults. In particular, we proposed that adult heartworm likely adopted an anaerobic electron transfer-based energy generation system distinct from the aerobic pathway utilized by its mammalian host, making it a promising target in developing next generation macrofilaricides and other treatments. Our survey provided novel insights into the D. immitis transcriptome and laid a foundation for further comparative studies on biology, parasitism and evolution within the phylum Nematoda. PMID- 16697389 TI - Chemical characterization of odorous gases at a landfill site by gas chromatography-mass spectrometry. AB - The composition of odorous gases emitted from a municipal landfill in the city of Izmir, Turkey was investigated using gas chromatography-mass spectrometry, and these data were examined in relation with the odor concentrations. Several volatile organic compounds (VOCs) were identified and quantified at five sampling sites in May and September 2005. Detected VOCs were monoaromatics (0.09-47.42 microg m(-3)), halogenated compounds (0.001-62.91microg m(-3)), aldehydes (0.01 38.55 microg m(-3)), esters (0.01-7.54 microg m(-3)), ketones (0.03-67.60 microg m(-3)), sulfur/nitrogen containing compounds (0.03-5.05 microg m(-3)), and volatile fatty acids (VFAs) (0.05-43.71 microg m(-3)). High levels of aldehydes (propanal up to 38.55 microg m(-3)) and VFAs (formic acid up to 43.71 microg m( 3)) were measured in May. However, VOC concentrations were relatively low in September. The monoaromatics and halogenated compounds were the abundant VOCs in landfill air for the both sampling periods. The benzene-to-toluene (B:T) ratio at the landfill site was significantly lower than urban areas indicating the presence of higher amounts of toluene in landfills compared to traffic exhaust rich urban areas. A statistically significant linear relationship was found between odor concentrations determined by olfactometry and total VOC concentrations. The relationships of odor concentrations with the different groups of chemicals were also examined using a step-wise multiple regression analysis. It was found that the concentrations of aldehydes, ketones, and esters are the best estimators, explaining 96% of the variability in odor concentrations (r2 = 0.96, n = 10, P < 0.01). PMID- 16697390 TI - The art and science of forming packed analytical high-performance liquid chromatography columns. AB - Columns of packed particles still are the most popular devices for high performance liquid chromatography (HPLC) separations because of their great utility, excellent performance and wide variety. However, the forming of packed beds for efficient, stable columns traditionally has been an art where the basics of how to form optimum beds generally was not well understood. The recent development of monolith rods was introduced in part to overcome the difficulty of producing stable beds of packing particles. However, these materials are less versatile than packed particle columns. Technology developments in recent years have produced a better understanding among those skilled in the practice of how to form optimized packed beds, and this has led to widely available, high-quality commercial columns. This presentation discusses the developments that led to the present state of column packing technology. Important steps in the packing of efficient, stable beds are described. The key step of selecting the best solvent for the slurry packing method is emphasized. Factors affecting the mechanical stability of packed columns also are discussed. The early art of packing columns now has evolved into a more scientific approach that allows the packing of good columns with a minimum of effort and time. PMID- 16697391 TI - Value of high-performance liquid chromatographic analysis of amino acids in the determination of Panax ginseng radix extract effect in cultured neurons. AB - The present research describes a reversed-phase high-performance liquid chromatographic (RP-HPLC) method that allows the determination of several amino acids in primary cultured cortical neurons of rats. The concentration of amino acids was determined by using pre-column derivatization with dansyl chloride and UV-diode array detection. Data show that Panax ginseng radix extract (GS) can modulate amino acid release in neurons. The levels of glutamate (Glu), aspartate (Asp), gamma-aminobutyric acid (GABA) and glycine (Gly) in the GS-treated groups were higher than in the non-treated groups dose-dependentwise. In this case, Glu and GABA were the most released amino acids (74.43% +/- 0.97 and 88.41% +/- 4.12 at ginseng dose 0.01 mg/ml after 1h from treatment, respectively). The values obtained in the determination of the analytical parameters (linearity, precision, limit of detection and accuracy) confirm the quality of the method. The average recoveries for intra and inter-day assay (n = 5) were 101.18 and 102.38 for Asp, 99.35 and 98.44 for Glu, 99.59 and 99.66 for Gly, and 100.06 and 100.37 for GABA. These data proved that the method yields accurate results, with RSD lower than 2.2%. The precision of the method was estimated on the basis of RSD of six injections at two different concentrations of amino acids. This technique is useful in studying the GS-mediated modulation of the dynamic equilibrium of amino acids and neurotransmission in neurons. PMID- 16697392 TI - Easy preparation and characterization of highly fluorescent polymer composite microspheres from aqueous CdTe nanocrystals. AB - Fluorescent microspheres were easily fabricated from aqueous CdTe nanocrystals (NCs). The NCs, which had negative charges on the surface, were first extracted to chloroform by cationic surfactant octadecyl-p-vinyl-benzyldimethylammonium chloride (OVDAC) and then swollen into performed polystyrene (PS) microspheres. Through this method, strong photoluminescence (PL) of aqueous NCs was inherited in the resultant composite microspheres. Moreover, the NCs were firmly stabilized in the microspheres, withstanding not only polar solvents but also nonpolar solvents. UV-vis spectrum, PL spectrum, TEM, and confocal fluorescence microscopy were used to characterize the product. PMID- 16697393 TI - Dependence of fragmentation behavior of colloidal aggregates on their fractal structure. AB - The fragmentation dynamics of aggregate of non-Brownian particles in shear flow is investigated numerically. The breakup behaviors of aggregates having the same connectivity but the different space-filling properties are examined. The Lagrangian particle simulation in a linear flow field is performed. The effect of surrounding fluid on the motion of multiple particles is estimated by Stokesian dynamics approach. The inter-particle force is calculated from the retarded van der Waals potential based on the Lifshitz theory. The results obtained in this work indicate that the fragmentation behavior of colloidal aggregates depends on their fractal structure. However, if the resultant aggregate size is smaller than the critical one, the fragmentation behavior shows the universality regardless of their original structure. Furthermore, the restructuring of aggregate in shear flow and its effect on the fragmentation process are also discussed. PMID- 16697394 TI - Safety of exercise testing in the chest pain unit: 31-mm ST elevation in variant angina. AB - We discuss a patient who presented with symptoms classic for variant angina with dramatic 31-mm ST elevation secondary to exercise testing in the chest pain unit, in whom neither myocardial infarction nor severe arrhythmia resulted. Although exercise testing is deemed generally safe, it has not been studied for safety per se in patients with variant angina. Further studies are needed to determine if the magnitude of ST elevation during exercise testing carries prognostic significance. PMID- 16697395 TI - Computer error and acceptable standards for electrocardiographic interpretation. PMID- 16697396 TI - Impulse propagation in late-stage embryonic and neonatal murine ventricular slices. AB - AIM: To evaluate whether field potential recordings from murine ventricular slice preparations serve as a model to investigate impulse propagation. METHOD: Late stage embryonic and neonatal murine hearts were sliced by a vibratome. Slices were placed on planar microelectrode arrays (MEAs). Field potentials of spontaneously beating and electrically stimulated contractions were recorded. The maximal negative deflection of the field potentials (dV / dt) was calculated to assess the local activation time, to create activation sequence maps, and to estimate conduction velocity. RESULTS: Mapping of impulse propagation of late stage embryonic and neonatal murine ventricular slices and estimation of conduction velocities is feasible using the MEA technique showing an impulse propagation reflecting anatomical structures and conduction velocities similar to those obtained with other techniques. CONCLUSION: The combination of viable ventricular slice preparations with the MEA technique offers a versatile and powerful technique to study cardiac impulse propagation. PMID- 16697397 TI - A specialized group of non-physician over-readers can expedite the final reporting process of the electrocardiogram. PMID- 16697398 TI - A technicians' perspective: tips and techniques for using disposable tab electrocardiogram electrodes; an easy to remember method for preventing electrocardiogram lead interchange. PMID- 16697399 TI - Bradycardia-dependent block in the accessory pathway in a patient with alternans Wolff-Parkinson-White syndrome. PMID- 16697400 TI - Diagnosis of inferior myocardial infarction based on the paced QRS complex. PMID- 16697401 TI - Recording lead V(4)R is associated to enhanced use of fibrinolytic therapy in acute myocardial infarction. AB - BACKGROUND: ST-segment elevation in the right-sided chest lead V(4)R in inferior wall myocardial infarction is recognized as a sign of proximal occlusion of the right coronary artery with evolving right ventricular myocardial infarction. Our objective is to study how often lead V(4)R is recorded in clinical practice and how this might be associated with use of reperfusion therapy and outcome of patients. METHODS: Recording of lead V(4)R in 814 consecutive patients with acute myocardial infarction, administration of therapy, and outcome of the patients during a median follow-up of 285 days (174-313 days) were studied. RESULTS: V(4)R was recorded in 52% of patients with inferior ST-elevation myocardial infarction. Patients with V(4)R recorded were more likely to receive fibrinolytic therapy compared with patients without recording (65% vs 51%; P = .035). In multivariate analysis, recording of lead V(4)R (odds ratio [OR] 1.6, 95% confidence interval [CI] 1.1-2.2; P = .006), along with age (P < .001), previous myocardial infarction (OR 2.2, 95% CI 1.3-3.5; P = .002), and diabetes (OR 3.9, 95% CI 1.1 2.4; P = .03) correlated to the use of reperfusion therapy. Patients with lead V(4)R recorded had less (P = .055) reinfarction, unstable angina, stroke, and/or death during follow-up. CONCLUSIONS: Lead V(4)R was recorded in only half of patients with inferior ST-elevation myocardial infarction. Patients with V(4)R recorded were more likely to receive thrombolytic therapy than those without recording of the additional chest lead. PMID- 16697402 TI - Tachyarrhythmias in percutaneous coronary interventions. AB - Accompanying the clear benefits, there are certain risks of tachyarrhythmias in percutaneous coronary interventions (PCI), including serious ventricular arrhythmias and atrial fibrillation (AF). Ventricular arrhythmias may result from excess catheter manipulation, intracoronary dye injection, new ischemic events, or reperfusion. In patients with heart failure such kind of arrhythmias can occur more frequently. Atrial dysfunction, sino-atrial and nodal ischemia, congestive heart failure, sympathetic stimulation, iatrogenic factors are the possible causes of AF especially in patients undergoing primary PCI. Atrial fibrillation, on the other hand, can cause clinical squeal in the setting of a rapid ventricular response or if the loss of atrial systole results in hypotension, as in a patient with mitral stenosis or diastolic ventricular dysfunction. Majority of the ventricular arrhythmias and AF tend to revert spontaneously. However, the special treatment must be given, when necessary. PMID- 16697403 TI - Electrocardiographic diagnosis of non-ST-segment elevation acute coronary syndromes: current concepts for the physician. AB - With several myocardial infarction (MI) registries reporting a decline in the incidence of ST-elevation MI (STEMI) and an increase in non-ST-elevation MI (NSTEMI) and unstable angina (UA), it is important that future healthcare resources are directed towards this increased volume of patients, ECG technology, core to the early diagnosis of these patients, has lagged behind relative to other techniques and little progress has been as far as acute coronary syndrome triage is concerned beyond ST-segment deviation. We present a review of the literature on current electrocardiographic changes which will allow admitting physicians to better risk stratify those patients with "non-diagnostic ECGs." These ECGs may become diagnostic with careful evaluation, use of serial ECGs and when additional lead sets are used. PMID- 16697404 TI - Urocortin inhibits Beclin1-mediated autophagic cell death in cardiac myocytes exposed to ischaemia/reperfusion injury. AB - Autophagy is known to be a feature of cardiomyopathies and chronic ischaemia. Here we demonstrate that autophagy is also induced by a single cycle of ischaemia/reperfusion (I/R in neonatal and adult rat cardiac myocytes). Consistent with the critical role for Beclin1 in autophagocytosis, reduction of Beclin1 expression in cardiac myocytes by RNAi reduces I/R-induced autophagy and this is associated with enhanced cell survival. Autophagy is also reduced by urocortin, an endogenous cardiac peptide which we have previously shown to reduce other forms of myocyte cell death induced by I/R. The inhibition of autophagy by urocortin is mediated in part by inhibition of Beclin1 expression, an effect which is mediated by activation of the PI3 kinase/Akt pathway but which does not involve activation of p42/p44 MAPK. PMID- 16697405 TI - Two-dimensional averaged images of the dynactin complex revealed by single particle analysis. AB - The dynactin complex interacts with dynein and numerous other proteins to provide for a wide range of subcellular transport functions. A detailed understanding of the structure and subunit organization of dynactin should yield new insights into its function. In the present study, we used single particle analysis to obtain a two-dimensional averaged image of dynactin isolated from chick embryo brains and visualized by negative stain electron microscopy (EM). Each individual image, consisting of the shoulder/sidearm and the rod, closely resembled the previously published quick-freeze deep-etch rotary-shadow electron micrographs. However, the averaged image revealed novel structural features that may have functional significance. The bulky shoulder complex has a triangular shape and is 13 nm wide and 8 nm high. The rod, with an overall length of 40 nm, consists of clearly defined lobes that are apparently grouped into three parts, the pointed-end complex, the middle segment, and the extra lobes at the barbed end. The pointed end complex reveals the characteristic protrusions and clefts that were previously observed only in the isolated pointed-end complex. In the middle segment, the seven lobes are fitted to the helical symmetry of F-actin. A narrow but prominent gap separates the previously unidentified extra three lobes at the barbed end from the middle segment. The averaged image we obtained contrasts dramatically with the simple Arp1 polymer that was previously reported by single particle analysis of bovine brain dynactin. These apparent structural differences are probably due to the greater stability and integrity of the chick embryo brain dynactin preparation. We propose a new structural model for dynactin, based on our observations. PMID- 16697406 TI - Quantitative analysis of multi-component spherical virus assembly: scaffolding protein contributes to the global stability of phage P22 procapsids. AB - Assembly of the hundreds of subunits required to form an icosahedral virus must proceed with exquisite fidelity, and is a paradigm for the self-organization of complex macromolecular structures. However, the mechanism for capsid assembly is not completely understood for any virus. Here we have investigated the in vitro assembly of phage P22 procapsids using a quantitative model specifically developed to analyze assembly of spherical viruses. Phage P22 procapsids are the product of the co-assembly of 420 molecules of coat protein and approximately 100 300 molecules of scaffolding protein. Scaffolding protein serves as an assembly chaperone and is not part of the final mature capsid, but is essential for proper procapsid assembly. Here we show that scaffolding protein also affects the thermodynamics of assembly, and for the first time this quantitative analysis has been performed on a virus composed of more than one type of protein subunit. Purified coat and scaffolding proteins were mixed in varying ratios in vitro to form procapsids. The reactions were allowed to reach equilibrium and the proportion of the input protein assembled into procapsids or remaining as free subunits was determined by size exclusion chromatography and SDS-PAGE. The results were used to calculate the free energy contributions for individual coat and scaffolding proteins. Each coat protein subunit was found to contribute 7.2(+/-0.1)kcal/mol and each scaffolding protein -6.1(+/-0.2)kcal/mol to the stability of the procapsid. Because each protein interacts with two or more neighbors, the pair-wise energies are even less. The weak protein interactions observed in the assembly of procapsids are likely important in the control of nucleation, since an increase in affinity between coat and scaffolding proteins can lead to kinetic traps caused by the formation of too many nuclei. In addition, we find that adjusting the molar ratio of scaffolding to coat protein can alter the assembly product. When the scaffolding protein concentration is low relative to coat protein, there is a correspondingly low yield of proper procapsids. When the relative concentration is very high, too many nuclei form, leading to kinetically trapped assembly intermediates. PMID- 16697407 TI - Human transcription factors contain a high fraction of intrinsically disordered regions essential for transcriptional regulation. AB - Human transcriptional regulation factors, such as activators, repressors, and enhancer-binding factors are quite different from their prokaryotic counterparts in two respects: the average sequence in human is more than twice as long as that in prokaryotes, while the fraction of sequence aligned to domains of known structure is 31% in human transcription factors (TFs), less than half of that in bacterial TFs (72%). Intrinsically disordered (ID) regions were identified by a disorder-prediction program, and were found to be in good agreement with available experimental data. Analysis of 401 human TFs with experimental evidence from the Swiss-Prot database showed that as high as 49% of the entire sequence of human TFs is occupied by ID regions. More than half of the human TFs consist of a small DNA binding domain (DBD) and long ID regions frequently sandwiching unassigned regions. The remaining TFs have structural domains in addition to DBDs and ID regions. Experimental studies, particularly those with NMR, revealed that the transactivation domains in unbound TFs are usually unstructured, but become structured upon binding to their partners. The sequences of human and mouse TF orthologues are 90.5% identical despite a high incidence of ID regions, probably reflecting important functional roles played by ID regions. In general ID regions occupy a high fraction in TFs of eukaryotes, but not in prokaryotes. Implications of this dichotomy are discussed in connection with their functional roles in transcriptional regulation and evolution. PMID- 16697408 TI - Identification of the PXW sequence as a structural gatekeeper of the headpiece C terminal subdomain fold. AB - The HeadPiece (HP) domain, present in several F-actin-binding multi-domain proteins, features a well-conserved, solvent-exposed PXWK motif in its C-terminal subdomain. The latter is an autonomously folding subunit comprised of three alpha helices organised around a hydrophobic core, with the sequence motif preceding the last helix. We report the contributions of each conserved residue in the PXWK motif to human villin HP function and structure, as well as the structural implications of the naturally occurring Pro to Ala mutation in dematin HP. NMR shift perturbation mapping reveals that substitution of each residue by Ala induces only minor, local perturbations in the full villin HP structure. CD spectroscopic thermal analysis, however, shows that the Pro and Trp residues in the PXWK motif afford stabilising interactions. This indicates that, in addition to the residues in the hydrophobic core, the Trp-Pro stacking within the motif contributes to HP stability. This is reinforced by our data on isolated C terminal HP subdomains where the Pro is also essential for structure formation, since the villin, but not the dematin, C-terminal subdomain is structured. Proper folding can be induced in the dematin C-terminal subdomain by exchanging the Ala for Pro. Conversely, the reverse substitution in the villin C-terminal subdomain leads to loss of structure. Thus, we demonstrate a crucial role for this proline residue in structural stability and folding potential of HP (sub)domains consistent with Pro-Trp stacking as a more general determinant of protein stability. PMID- 16697410 TI - Docking of a single phage lambda to its membrane receptor maltoporin as a time resolved event. AB - We have been able to observe the first step in bacteriophage infection, the docking of phage lambda to its membrane receptor maltoporin, at the single particle level. High-resolution conductance recording from a single trimeric maltoporin channel reconstituted into a planar lipid bilayer has allowed detection of the simultaneous and irreversible interaction of the phage tail with all three monomers of the receptor. The formation of a phage-maltoporin complex affects the channel transport properties. Our analysis demonstrates that phage attaches symmetrically to all three receptor monomers. The statistics of sugar binding to the phage-receptor complex on the side opposite to phage docking show that the monomers of maltoporin still bind sugar independently, with the kinetic constants expected from those of the phage-free receptor. This finding suggests that phage docking does not distort the structure of the receptor, and that the phage-binding regions are close to, but do not overlap with, the sugar-binding domains of the maltoporin monomers. However, ion fluxes through the pores of maltoporin in the phage-receptor complex share a new common pathway. We expect that the present study contributes to the current needs for structural information on the functional complexes involved in intercellular recognition. PMID- 16697409 TI - Evolutionarily distinct residues in the uncoupling protein UCP1 are essential for its characteristic basal proton conductance. AB - The uncoupling proteins (UCPs) are mitochondrial transporters that modulate the efficiency of oxidative phosphorylation. Members of this family have been described in many phyla within the animal and plant kingdoms, as well as in fungi. The mammalian uncoupling protein UCP1 is activated by fatty acids and inhibited by nucleotides. In the absence of both regulators, UCP1 presents a high ohmic proton conductance that is a unique property of this carrier. The increasing number of protein sequences available has enabled us to apply a sequence analysis approach to investigate transporter function. We reconstructed a robust phylogeny of UCPs and used comparative sequence analysis to search for phylogenetically shared derived sequence features that may confer distinct properties on UCP1. We assessed the functional relevance of shared derived UCP1 residues by substituting them with their counterparts in UCP2, and expressing the protein chimeras in yeast. We found that substitution of both Glu134 and Met140 abolishes the basal proton permeability of UCP1 while preserving fatty acid activation and its nucleotide inhibition. PMID- 16697411 TI - The thioesterase domain of the fengycin biosynthesis cluster: a structural base for the macrocyclization of a non-ribosomal lipopeptide. AB - Many secondary metabolic peptides from bacteria and fungi are produced by non ribosomal peptide synthetases (NRPS) where the final step of biosynthesis is often catalysed by designated thioesterase domains. Here, we report the 1.8A crystal structure of the fengycin thioesterase (FenTE) from Bacillus subtilis F29 3, which catalyses the regio- and stereoselective release and macrocyclization of the antibiotic fengycin from the NRPS template. A structure of the PMSF inactivated FenTE domain suggests the location of the oxyanion hole and the binding site of the C-terminal residue l-Ile11 of the lipopeptide. Using a combination of docking, molecular dynamics simulations and in vitro activity assays, a model of the FenTE-fengycin complex was derived in which peptide cyclization requires strategic interactions with residues lining the active site canyon. PMID- 16697412 TI - Psychiatric syndromes comorbid with mental retardation: differences in cognitive and adaptive skills. AB - The study concerns the specific cognitive and adaptive skills of persons dually diagnosed with mental retardation (MR) and comorbid pathologies, as schizophrenia, personality and mood disorders, pervasive developmental disorders, epilepsy and ADHD. The sample was composed of 182 subjects, diagnosed as mild or moderate MR level, age range from 6 years 8 months to 50 years 2 months, mean age 17.1 (standard deviation 7.9). All the subjects were inpatients in a specialized structure for the diagnosis and the treatment of MR. The instruments of the study were Wechsler Intelligence Scale (WAIS-R or WISC-R according to the chronological age of subjects) and Vineland Adaptive Behavior Scale (VABS). Results confirm that comorbidity is a factor differentiating among mentally retarded subjects. Both verbal processes requiring memory retrieval and visuo-spatial processes are involved as differentiating features. ADHD strongly increases the impairment of cognitive skills, while behavioral disorders are less damaging in MR performance. In adult samples, the differentiating role of comorbid syndromes in MR individuals is reduced for cognitive skills, and limited to some basic verbal abilities, more impaired in mood disorder, less in schizophrenic disorder. The areas of adaptation and socialization, motor and daily living skills, are impaired more in generalized development disturbances than in comorbid schizophrenic and personality and mood disorders. An accurate psychological assessment of dual diagnoses is useful in detecting the specific underlying processes differentiating the comorbid syndromes, and in planning an appropriate rehabilitative treatment. PMID- 16697414 TI - Nonfatal injuries following Hurricane Katrina--New Orleans, Louisiana, 2005. AB - The Journal of Safety Research has partnered with the National Center for Injury Prevention and Control at the Centers for Disease Control and Prevention (CDC) in Atlanta, Georgia, USA, to briefly report on some of the latest findings in the research community. This report is the fourth edition in a series of CDC articles. BACKGROUND: An active injury and illness surveillance system was established by the Centers for Disease Control and Prevention (CDC) along with the Louisiana Department of Health and Hospitals (LDHH) in the aftermath of Hurricane Katrina in functioning hospitals and medical clinics. RESULTS: The surveillance system recorded 7,543 nonfatal injuries among residents and relief workers between September 8-October 14, 2005. The leading mechanisms of injury identified in both groups were fall and cut/stab/pierce, with a greater proportion of residents compared to relief workers injured during the repopulation period. Clean-up was the most common activity at the time of injury for both groups. CONCLUSION: Injuries documented through this system underscore the need for surveillance of exposed populations to determine the injury burden and initiate injury prevention activities and health communication campaigns. PMID- 16697413 TI - Estrogen and response to sertraline in postmenopausal women with major depressive disorder: a pilot study. AB - OBJECTIVE: Pilot study examining the effects of estrogen therapy (ET) on antidepressant response in postmenopausal women with major depressive disorder (MDD). METHODS: Twenty-two subjects received sertraline at 50mg/day for one week, with an increase to 100mg/day at week 2 for a 10-week trial. Transdermal estrogen or placebo patches 0.1mg were randomly administered concurrent with the initiation of sertraline treatment. The 21 item Hamilton Depression Rating Scale (HDRS-21) was administered to all patients at baseline and weekly thereafter. RESULTS: Both groups showed a similar significant reduction in HDRS-21 scores by the end of the study. There was no significant difference between the two treatment groups at the end of the 10-week trial, but the women receiving sertraline with ET showed significantly greater early improvement (weeks 2-4) compared to the women receiving sertraline with placebo. CONCLUSIONS: Sertraline is an effective antidepressant for postmenopausal women with MDD. ET does not alter the response rate to antidepressant therapy however ET may play a role in accelerating the antidepressant response. PMID- 16697415 TI - Optical tissue window: a novel model for optimizing engraftment of intestinal stem cell organoids. AB - BACKGROUND: Intestinal malabsorption disorders and short bowel syndrome lead to significant morbidity. We recently demonstrated that grafting of intestinal organoids can grow a bioengineered intestinal neomucosa and cure bile acid malabsorption in rats. Now we have developed a novel system that permits direct observation of intestinal organoids in vivo to optimize conditions for engraftment. METHODS: Optical Windows were created in C57BL/6J mice by externalizing an omental pedicle into a dorsal skin flap chamber. Following creation of windows, 5000 intestinal organoids from green-fluorescent protein transgene (GFP)+ donor mice were seeded directly either on omentum or on polyglycolic acid (PGA) disks that had been placed on omentum at 1 or 5 days. Engraftment of green fluorescent cells was evaluated on postseeding days 1, 3, 5, 7, 10, 12, and 21 using fluorescence microscopy. RESULTS: An initial group had seeding onto omentum (n = 5) or biopolymer disks (n = 5) on postoperative day 1. After 7 days, there was mucosal cell engraftment onto omental tissue and biopolymers. GFP+ organoids engrafted significantly better when seeded onto biopolymers compared to omentum (P < 0.05). In a second study with increased sample size (n = 24) up to day 12, all four groups demonstrated adherence and growth. However, GFP+ organoids seeded onto delayed PGA biopolymer demonstrated significantly better engraftment (P < 0.05). CONCLUSIONS: This novel system allows continuous in vivo observation of engrafted cells that are seeded on externalized omentum. The use of PGA mesh biopolymer may improve engraftment of intestinal organoids. PMID- 16697416 TI - A probabilistic approach to compact steady-state kinetic equations for enzymic reactions. AB - We describe a method for deriving kinetic equations based on the simplification of a complex graphical scheme of steady-state enzymic reactions to one that is comprised of an unbranched pathway. It entails compressing unbranched multi-step sequences into one step, and fusing some graph nodes into a single node. The final form of the equations is compact and well structured, and it simplifies the choice of independent kinetic parameters. The approach is illustrated by an analysis of representative two- and three-substrate reactions. PMID- 16697417 TI - Simple mathematical model of pathologic microsatellite expansions: when self reparation does not work. AB - We propose a simple model of pathologic microsatellite expansion, and describe an inherent self-repairing mechanism working against expansion. We prove that if the probabilities of elementary expansions and contractions are equal, microsatellite expansions are always self-repairing. If these probabilities are different, self reparation does not work. Mosaicism, anticipation and reverse mutation cases are discussed in the framework of the model. We explain these phenomena and provide some theoretical evidence for their properties, for example, the rarity of reverse mutations. PMID- 16697418 TI - p38 and ERK MAP kinase mediates iron chelator-induced apoptosis and -suppressed differentiation of immortalized and malignant human oral keratinocytes. AB - Iron is essential for neoplastic cell growth, and iron chelators have been tested for potential anti-proliferative and anti-cancer effects, but the effects of iron chelators on oral cancer have not been clearly elucidated. To determine the mechanism of cell death induced by iron chelators, we explored the pathways of the three structurally related mitogen-activated protein (MAP) kinase subfamilies during iron chelator-induced apoptosis and differentiation of immortalized human oral keratinocytes (IHOK) and oral cancer cells (HN4). The iron chelator deferoxamine (DFO) exerted potent time- and dose-dependent inhibitory effects on the growth and apoptosis of IHOK and HN4 cells. DFO strongly activates p38 MAP kinase and extracellular signal-regulated kinase (ERK), but does not activate c Jun N-terminal kinase/stress-activated protein kinase. Of the three MAP kinase blockers used, the selective p38 MAP kinase inhibitor SB203580 and ERK inhibitor PD98059 protected IHOK and HN4 cells against iron chelator-induced cell death, which indicates that the p38 and ERK MAP kinase is a major mediator of apoptosis induced by this iron chelator. Interestingly, treatment of IHOK and HN4 cells with SB203580 and PD98059 abolished cytochrome c release, as well as the activation of caspase-3 and caspase-8. DFO suppressed the expression of epithelial differentiation markers such as involucrin, CK6, and CK19, and this suppression was blocked by p38 and ERK MAP kinase inhibitors. Collectively, these data suggested that p38 and ERK MAP kinase plays an important role in iron chelator-mediated cell death and in the suppression of differentiation of oral immortalized and malignant keratinocytes, by activating a downstream apoptotic cascade that executes the cell death pathway. PMID- 16697419 TI - Gender-specific orexigenic and anorexigenic mechanisms in rats. AB - Feeding dysregulation may manifest as either under-nourishment (e.g., anorexia) or excessive eating leading to obesity. Recent studies have suggested a gender related variance in weight maintenance in response to chronic disease or obesity related dietary regimens. However it is unclear whether these gender differences in weight management are secondary to appetite-mediated food intake or alternative mechanisms (e.g., exercise, metabolism). In this study, we explored gender-dependent feeding and hormonal responses to dietary restriction (12-h fast) or to an inflammatory stimulus (LPS, 100 microg/kg b.w.; i.p.) in rats. In response to a 12 h fast, female rats increased (p<0.05) total daily food intake above that of male rats by primarily increasing nighttime feeding by 40%, as compared to 10% in males. Consistent with the increased food intake, fasting induced a greater percent increase in female as compared to male plasma ghrelin (141 vs. 65%, p<0.001). In response to LPS, both male and female rats showed similar reductions in total daily food consumption. However LPS (6 h) induced a greater percent increase in plasma leptin in female than male rats (230 vs. 33%, p<0.01), whereas ghrelin was similarly decreased in both females and males (66 vs. 44%). These findings demonstrate sexual dimorphic responses in feeding and appetite-associated hormonal responses to fasting or LPS treatment. Our findings suggest that therapeutic interventions with ghrelin or leptin must be modified according to gender in order to optimally achieve either weight loss for obesity or weight gain/maintenance for chronic illness-associated anorexia. PMID- 16697420 TI - The occurrence of imposex in Hexaplex trunculus from the Croatian Adriatic. PMID- 16697421 TI - The Prestige oil spill: a scientific response. PMID- 16697422 TI - Antagonistic activity of one-joint muscles in three-dimensions using non-linear optimisation. AB - Non-linear optimisation, such as the type presented by R.D. Crowninshield and R.A. Brand [The prediction of forces in joint structures: Distribution of intersegmental resultants, Exercise Sports Sci. Rev. 9 (1981) 159], has been frequently used to obtain a unique set of muscle forces during human or animal movements. In the past, analytical solutions of this optimisation problem have been presented for single degree-of-freedom models, and planar models with a specific number of muscles and a defined musculoskeletal geometry. Results of these studies have been generalised to three-dimensional problems and for general formulations of the musculoskeletal geometry without corresponding proofs. Here, we extend the general solution of the above non-linear, constrained, planar optimisation problem to three-dimensional systems of arbitrary geometry. We show that there always exists a set of intersegmental moments for which the given static optimisation formulation will predict co-contraction of a pair of antagonistic muscles unless they are exact antagonists. Furthermore, we provide, for a given three-dimensional system consisting of single joint muscles, a method that describes all the possible joint moments that give co-contraction for a given pair of antagonistic muscles. PMID- 16697423 TI - Some factors affecting pulmonary oxygen transport. AB - Equations governing oxygen transport from alveolar gas to red blood cells flowing through pulmonary capillaries are written down. Some analytical predictions are made on factors affecting the rate at which this process takes place. Numerical simulations are carried out to investigate the effect of red blood cell shape, capillary dimensions, haematocrit and choice of oxygen dissociation curve on pulmonary oxygen transport. These factors all have an effect on pulmonary transport, with the effect being much more marked for simulations with low oxygen levels, typical of those seen in some subjects with respiratory disease. PMID- 16697424 TI - Modelling the balance between quiescence and cell death in normal and tumour cell populations. AB - When considering either human adult tissues (in vivo) or cell cultures (in vitro), cell number is regulated by the relationship between quiescent cells, proliferating cells, cell death and other controls of cell cycle duration. By formulating a mathematical description we see that even small alterations of this relationship may cause a non-growing population to start growing with doubling times characteristic of human tumours. Our model consists of two age structured partial differential equations for the proliferating and quiescent cell compartments. Model parameters are death rates from and transition rates between these compartments. The partial differential equations can be solved for the steady-age distributions, giving the distribution of the cells through the cell cycle, dependent on specific model parameter values. Appropriate formulas can then be derived for various population characteristic quantities such as labelling index, proliferation fraction, doubling time and potential doubling time of the cell population. Such characteristic quantities can be estimated experimentally, although with decreasing precision from in vitro, to in vivo experimental systems and to the clinic. The model can be used to investigate the effects of a single alteration of either quiescence or cell death control on the growth of the whole population and the non-trivial dependence of the doubling time and other observable quantities on particular underlying cell cycle scenarios of death and quiescence. The model indicates that tumour evolution in vivo is a sequence of steady-states, each characterised by particular death and quiescence rate functions. We suggest that a key passage of carcinogenesis is a loss of the communication between quiescence, death and cell cycle machineries, causing a defect in their precise, cell cycle dependent relationship. PMID- 16697425 TI - Mid- to late gestational morphine exposure does not alter the rewarding properties of morphine in adult male rats. AB - Prenatal exposure to drugs of abuse often leads to physiological and neurobiological abnormalities including decreased brain and body weight, cognitive deficits and behavioral alterations. A handful of studies showed increased vulnerability to drug abuse in prenatally drug-exposed offspring. Our work also demonstrated that prenatal exposure to analgesic doses of morphine during gestation days 11-18 increases mu-opioid receptor density in the nucleus accumbens and central amygdala of adult male rats. Both the nucleus accumbens and central amygdala play important roles in modulating drug-induced reward via the mesolimbic dopaminergic system. Therefore, two types of behavioral paradigms were used to test the hypothesis that the same prenatal morphine exposure would enhance the rewarding effects of morphine, making drug-exposed offspring more vulnerable to abuse this drug in adulthood. All experiments were performed with adult male offspring of saline-injected, morphine-injected or non-injected (control) dams. (1) The unbiased conditioned place preference (CPP) paradigm was used to investigate whether prenatal morphine exposure sensitizes adult male rats to non-contingent morphine reward. These adult animals were conditioned with 0.1, 0.3, 1, 3 or 5 mg/kg morphine. All control, prenatally saline- and morphine exposed male rats preferred the morphine-paired compartment relative to the saline-paired compartment. However, the magnitude of morphine CPP in adult male rats was not dependent on the conditioning dose of morphine or prenatal morphine exposure. (2) Intravenous morphine self-administration was used to assess the behavioral response to contingent morphine reward. Each rat self-administered one of four doses of morphine (0.3, 1, 2 or 3 mg/kg/infusion). Morphine self administration was not altered in prenatally morphine-exposed adult male offspring. Control males self-administered significantly less morphine at the lowest dose of morphine than both prenatally saline- and morphine-exposed males. Although our data show that prenatal exposure to an analgesic dose of morphine during the time of opioid receptor appearance does not enhance morphine CPP or self-administration, they do not exclude the possibility that this prenatal morphine exposure enhances the rewarding properties of other drugs of abuse. PMID- 16697426 TI - Retigabine and flupirtine exert neuroprotective actions in organotypic hippocampal cultures. AB - Retigabine and flupirtine are two structurally related molecules provided of anticonvulsant and analgesic actions. The present study has investigated the neuroprotective potential, as well as the possible underlying molecular mechanisms, exerted by retigabine and flupirtine in rat organotypic hippocampal slice cultures (OHSCs) exposed to N-methyl-D-aspartate (NMDA), oxygen and glucose deprivation followed by reoxygenation (OGD), or serum withdrawal (SW). Region specific vulnerability of hippocampal subfields occurred with each of these injury models. Specifically, CA1 was the most susceptible region to both NMDA and OGD-induced neurodegeneration, whereas selective cell death in the dentate gyrus (DG) occurred upon OHSCs exposure to SW. The NMDA antagonist MK-801 (10-30 microM), despite blocking NMDA- and OGD-induced cell death, failed to prevent SW induced neurodegeneration. Interestingly, retigabine (0.01-10 microM) and flupirtine (0.01-10 microM) dose-dependently prevented DG neuronal death induced by SW, with IC50 s of 0.4 microM and 0.7 microM, respectively. By contrast, retigabine and flupirtine (each at 10 microM) were less effective in counteracting NMDA- or OGD-induced toxicity in the CA1 region. Both retigabine and flupirtine (0.1-10 microM) reduced SW-induced ROS production in the DG with IC50 s of approximately 1 microM. This suggested that antioxidant actions of these compounds participated in OHSC neuroprotection during SW. By contrast, activation of KCNQ K+ channels seemed not to be involved in retigabine-induced OHSCs neuroprotection during SW, since linopirdine (20 microM) and XE-991 (10 microM), two KCNQ blockers, failed to reverse retigabine-induced neuronal rescue. PMID- 16697427 TI - QF2004B, a potential antipsychotic butyrophenone derivative with similar pharmacological properties to clozapine. AB - The aim of the present work was to characterize a lead compound displaying relevant multi-target interactions, and with an in vivo behavioral profile predictive of atypical antipsychotic activity. Synthesis, molecular modeling and in vitro and in vivo pharmacological studies were carried out for 2-[4-(6 fluorobenzisoxazol-3-yl)piperidinyl]methyl-1,2,3,4-tetrahydro-carbazol-4-one (QF2004B), a conformationally constrained butyrophenone analogue. This compound showed a multi-receptor profile with affinities similar to those of clozapine for serotonin (5-HT2A, 5-HT1A, and 5-HT2C), dopamine (D1, D2, D3 and D4), alpha adrenergic (alpha1, alpha2), muscarinic (M1, M2) and histamine H1 receptors. In addition, QF2004B mirrored the antipsychotic activity and atypical profile of clozapine in a broad battery of in vivo tests including locomotor activity (ED50 = 1.19 mg/kg), apomorphine-induced stereotypies (ED50 = 0.75 mg/kg), catalepsy (ED50 = 2.13 mg/kg), apomorphine- and DOI (2,5-dimethoxy-4-iodoamphetamine) induced prepulse inhibition (PPI) tests. These results point to QF2004B as a new lead compound with a relevant multi-receptor interaction profile for the discovery and development of new antipsychotics. PMID- 16697428 TI - The relationship between working memory and episodic memory disorders in transient global amnesia. AB - In a previous study, we investigated the relationship between the disorders of both episodic memory and working memory in the acute phase of transient global amnesia (TGA). Since executive functions were spared, another dysfunction may be responsible for the binding and maintenance of multimodal informations and contribute to the encoding disorders observed in some patients [Quinette, P., Guillery, B., Desgranges, B., de la Sayette, V., Viader, F., & Eustache, F. (2003). Working memory and executive functions in transient global amnesia. Brain, 126, 1917-1934.]. The aim of this present study was to assess the functions of binding and maintenance of multimodal information during TGA and explore their involvement in episodic memory disorders. We therefore conducted a more thorough investigation of working memory in 16 new patients during the acute phase of TGA using two tasks designed to assess the binding process and both dimensions of the maintenance, namely the active storage and the memory load ability. We also investigated the nature of the episodic memory impairment in distinguishing between the performance of patients with preferential encoding deficits and those of patients with preferential storage disorders on the episodic memory task. This distinction was closely related to the severity of amnesia, i.e. an encoding disorder was observed rather in the early phase of TGA. The results showed that while the functions of binding and maintenance of multimodal information were intact in patients with storage disorders, they were impaired in the case of encoding deficits. These results are interpreted in the recent framework of episodic buffer proposed by Baddeley [Baddeley, A. D. (2000). The episodic buffer: A new component of working memory? Trends in Cognitive Sciences, 4, 417-423] that represents an interface between working memory and episodic memory. PMID- 16697429 TI - Knowing 7 x 8, but not the meaning of 'elephant': evidence for the dissociation between numerical and non-numerical semantic knowledge. AB - Patients affected by semantic dementia (SD) and other severe cognitive deficits may show preserved numerical skills, including the retrieval of multiplication facts from long-term memory. No studies so far specifically investigated the network of arithmetic facts in semantic dementia. Thus, it is unknown whether preserved multiplication in SD truly reflects intact semantic knowledge or preserved retrieval of verbal sequences (just as the recitation of rhymes or poems). In the present study a patient (SG) with SD underwent an extensive assessment of number processing and calculation abilities. In particular, multiplication knowledge was investigated through a series of computerised tasks (production task, multiple-choice task, number bisection task with multiplicative triplets, number-matching task). SG demonstrated excellent performance in all number processing and calculation tasks. In computerised tasks tapping multiplication fact knowledge, SG was as accurate and fast as healthy controls. Analyses on individual regression slopes indicated that SG's reaction time effects (problem-size effect, problem-difficulty effect, interference effects, and facilitation effect) were comparable to those found in controls in each task. These results add new evidence to the independence of numerical knowledge from other semantic information and provide further insight into the organisation of stored arithmetic knowledge. PMID- 16697430 TI - Unconscious processing of Arabic numerals in unilateral neglect. AB - This study explores the processing of Arabic numerals in three patients with dense left unilateral neglect. Three tasks have been used: a test of visual awareness (is the stimulus on the left, on the right or on both sides?), a number comparison task (is the number larger or smaller than 5?), a number parity judgment task (is the number odd or even?). The test of visual awareness indicated that all three patients were completely unaware of the stimuli presented in the left hemifield. Despite this, the number comparison and number parity judgment tasks clearly indicated that Arabic numerals were unconsciously processed at semantic level (i.e. quantity). These results show that patients with left unilateral neglect can still semantically process Arabic numerals unconsciously. PMID- 16697431 TI - Doppler evaluation of maternal and fetal vessels during normal gestation in the bitch. AB - The aim of this work is to evaluate the haemodynamic characteristics of maternal and fetal vessels during normal pregnancy in bitches, using Colour and Pulsed wave Doppler ultrasonography, in order to obtain more information about maternal and fetal circulation. The blood waveforms of the uteroplacental arteries, aorta, caudal cava vein and umbilical cord of the fetuses were recorded weekly in 16 pregnant bitches. Also, the measurements of Peak Systolic, End Diastolic Velocities, Resistance and Pulsatility Indices were carried out. Uteroplacental blood flow was biphasic while the ones of the umbilical artery and aorta were first systolic and then diastolic. The cava showed a typical waveform of venous vessels. During gestation the EDV and PSV of fetal vessels increased (alpha<0.05) while the PI and RI of all vessels examined decreased (alpha<0.05) except for the IP of the Aorta. The Doppler ultrasonography was used to study the characteristics of maternal and fetal vessel flow and their progressive changes during pregnancy. This study can be considered a further contribution in diagnosing and monitoring high-risk pregnancies in Veterinary Medicine. PMID- 16697432 TI - A simple validated prediction rule to diagnose typhoid fever in Turkey. AB - The goal of this study was to develop a simple prediction rule for the diagnosis of typhoid fever. A model for the prediction of patients with typhoid fever at hospital admission was derived and validated by assigning weighted point values to independent predictive factors associated with a diagnosis of typhoid fever at hospital admission. Patient demographic, clinical and laboratory variables were used to compare patients with blood culture-confirmed typhoid fever with patients with fever of unknown origin. The model was derived and validated in two separate cohorts of patients from Dicle University Hospital in Diyarbakir, Turkey. A total of 371 patients were enrolled. A diagnostic index score was created using seven independent predictive factors associated with typhoid fever at hospital admission: age <30 years, abdominal distention, confusion, leukopenia, relative bradycardia, positive Widal test and a typhoid tongue. A clinical prediction rule helped to differentiate patients with typhoid fever. PMID- 16697433 TI - Relationship between Rayleigh wave polarization and state of stress. AB - This research develops an analytical model (using Stroh's formalism) to predict the affect of applied stress on the wave speed and the polarization of Rayleigh surface waves. Simulation results are then used to demonstrate that the polarization of a Rayleigh wave (which is reference-free) could be more sensitive than wave speed as an indicator of the state of stress. PMID- 16697434 TI - Perceptual learning without feedback in non-stationary contexts: data and model. AB - The role of feedback in perceptual learning is probed in an orientation discrimination experiment under destabilizing non-stationary conditions, and explored in a neural-network model. Experimentally, perceptual learning was examined with periodic alteration of a strong external noise context. The speed of learning, the performance loss at each change in external noise context (switch cost), and the asymptotic accuracy d' without feedback were very similar or identical to those with feedback. However, lack of feedback led to higher decision bias (error responses matching the external noise context). In the model, the stimulus representations are constant, whereas the read-out connections to a decision unit learn by a Hebbian plasticity rule that may be augmented by additional feedback input and criterion control of decision bias. PMID- 16697435 TI - Effects of interactions among wave aberrations on optical image quality. AB - Wave aberrations degrade the optical quality of the eye relative to the diffraction limit, but there are situations in which having slightly aberrated optics can provide some relative visual benefits. This fact led us to consider whether interactions among aberrations in the eye's wavefront produce an advantage for image quality relative to wavefronts with randomized combinations of aberrations with the same total RMS error. Total ocular wave aberrations from two experimental groups and corneal wave aberrations from one group were measured and expressed as Zernike polynomial expansions through the seventh-order. In a series of Monte Carlo simulations, modulation transfer functions (MTFs) for the measured wave aberrations were compared to distributions of artificial MTFs for wavefronts created by randomizing the sign or orientation of the aberrations, while maintaining the RMS error within each Zernike order. In a control condition, "synthetic" model eyes were produced by choosing each individual aberration term at random from individuals in the experimental group, and again MTFs were compared for original and randomized signs. Results were summarized by the MTF ratio: real MTF/mean simulated MTF, as a function of spatial frequency. For a 6mm pupil, the mean MTF ratio for total ocular aberrations was greater than 1.0 up to 60 cycles per degree, suggesting that the eye's aberrations are not independent and that there may be a positive functional consequences to their interrelations. This positive relation did not hold for corneal aberrations alone, or for the synthetic eyes. PMID- 16697436 TI - Effect of blur adaptation on blur sensitivity in myopes. AB - Although blur adaptation in myopia has been investigated, knowledge regarding its effect on blur sensitivity remains unknown. In the present study, changes in three blur thresholds (i.e., noticeable, bothersome, and non-resolvable blur) were assessed monocularly after 1h of blur adaptation in myopes. A Badal optical system was used to present either an isolated 20/50 Snellen E or 20/50 lines of text, with the full text field used in the latter condition for all blur judgments. Eight visually normal adult myopes were tested with paralyzed accommodation. All subjects exhibited blur adaptation, with a significant improvement in group mean visual acuity of -0.16 LogMAR. There was a consistent and concurrent significant decrease of 0.15-0.19 D in all blur thresholds for the isolated 20/50 E. However, there was no significant effect of blur adaptation on blur thresholds for the 20/50 text, with large intersubject variability evident. The enhanced blur sensitivity for the isolated E target may in part be attributed to the increased visual resolution following blur adaptation. Differences found in the blur thresholds for the two targets may be related to a variety of neuroperceptual phenomena, in particular lateral masking. PMID- 16697437 TI - The size distribution of polychlorinated dibenzo-p-dioxins and dibenzofurans in the bottom ash of municipal solid waste incinerators. AB - In this study, bottom ash was sampled from two Taiwanese municipal solid waste incinerators (MSWIs: A and B) and sieved to size classes of 4.75-9.5 mm, 2.36 4.75 mm, 1.0-2.36 mm, 0.6-1.0 mm, 0.3-0.6 mm, 0.21-0.3 mm, 0.125-0.21 mm, 0.075 0.125 mm and <0.074 mm. For both MSWIs, the major peak in the particle size distribution for the PCDD/F content was found in a particle size <0.21 mm, that is, 16.1 (A) and 4.37 pg I-TEQ/g (B). This is due to the fact that a smaller particle has higher specific surface area thus offering more adsorption sites. The mean cumulative fractions (F%) of PCDD/F I-TEQ in the bottom ash of MSWI A and B in the particle size range below 0.6, 2.36 and 4.75 mm, in sequence, are 40.4%, 77.2% and 95.6%, respectively. We suggest sieving the bottom ash before the subsequent reutilization at the suggested cut size of 0.6 mm. In the view of PCDD/Fs, bottom ash with a particle size larger than 0.6 mm, which is the more non-hazardous part, may be suitable for being the raw material of landfilling soil, road sub-base, and construction blocks. For the minus 0.6 mm fraction, vitrification with fly ash is one of the choices. PMID- 16697438 TI - Accumulation of organochlorine pesticides by semipermeable membrane devices using composite complex. AB - Semipermeable membrane devices (SPMDs) were developed for passive in situ monitoring of organochlorine pesticides (OCPs) in aqueous solution in both laboratory and field (Pearl River Delta, China) studies. The device consisted of a thin film of neutral lipid triolein, enclosed in thin-walled tubing made of composite cellulose acetate membrane (CA) supported by linear low density polyethylene (LLDPE) (CAPE). Results from the laboratory and field application indicated that triolein-CAPE (TCAPE) could quickly and efficiently accumulate hydrophobic OCPs in water and uptake equilibrium could reached within 20h in the laboratory. Some mathematical relationships of TCAPE-water partition coefficient (logK(sw)), triolein-water partition coefficient (logK(tw)) and octanol-water partition coefficient (logK(ow)) were developed under the laboratory conditions. A good correlation of accumulation in TCAPE with r(2) values ranging from 0.55 to 0.86 for individual OCPs (n=8) and an excellent correlation of logK(sw) and logK(ow) was also obtained under the field conditions. The average OCPs concentration in the surface water could be estimated by measuring OCPs concentration in the device under the field conditions. PMID- 16697439 TI - Use and storage of domestic pesticides in the UK. AB - Several epidemiological studies, particularly from the United States, have suggested health concerns may arise from the chronic exposure of young children to pesticides in their home environment. Most of these studies are retrospective and the available pesticide exposure information is either subject to the problem of recall or derived from surrogate information. Currently, little is known about the extent of pesticide use in homes and gardens in the UK. This was investigated in this study using a sample of parents from the Avon Longitudinal Study of Parents and Children (ALSPAC) cohort in and around Bristol. The level and extent of pesticide use by parents was explored through interviews that took place at the homes of the participants during August to November 2001, followed by an inventory of products stored in the home. From the results, 93% of parents had used at least one pesticide over the preceding year with the highest use being in the garden, second inside the home, followed by treating pets and head lice. Most households reported using one to five different products over the past year, with a mean of 3.5. The frequency of pesticide use showed that a minority of parents are using pesticides in the high frequency range. Over 85 different pesticide products were found to be stored in 76% of homes, with 76 different types of pesticide active ingredients. Insecticides were the most commonly used and stored pesticide. PMID- 16697440 TI - Microbiological and chemical properties of litter from different chicken types and production systems. AB - Chicken litter is produced in large quantities from all types of poultry raising activities. It is primarily used for land application, thus it is essential to analyze its properties before it is released to the environment. The objective of this study is to compare the microbiological and chemical properties of litter generated from layer and broiler chickens reared under intensive and free-range production systems. The microbiological analysis consisted of the enumeration of total bacteria, total coliforms, Staphylococcus species, Salmonella species and Clostridium perfringens. Chicken litter from layers reared under intensive and free range systems showed lower mean total bacterial count than the litter collected from chicken broilers reared under either of the two systems (P=0.0291). The litter from intensive layers had the lowest mean total coliform counts (P=0.0222) while the lowest Staphylococcus species count was observed in the litter from free-range layers (P=0.0077). The C. perfringens count was the lowest in chicken litter from intensively raised broilers and layers (P=0.0001). The chemical properties of litter from the different chicken types and production systems were compared based on determination of pH, electrical conductivity, carbon, nitrogen, phosphorus, potassium, cadmium and zinc. Litter from free-range broilers showed the highest pH value (P=0.0005); however, the electrical conductivity was higher in the litter from both intensive and free-range layers compared to the litter from both broiler production systems (P=0.0117). Chicken litter from intensive systems had higher nitrogen content than litter from free range systems (P=0.0000). The total phosphorus was the lowest in free-range broiler litter (P=0.0001), while the total potassium was the lowest in litter from intensively managed broilers (P=0.0000). Zinc appeared higher in litter from layers compared to that from broilers (P=0.0101). The cadmium content was higher in the litter from free-range broilers and layers compared to that in the litter from intensively managed systems (P=0.0439). Staphylococcus species in the litter as well as cadmium concentrations seem to be the most critical parameters presenting risks on the environment and on human health. Based on the lowest coliform counts (an indication of water pollution), the high nutrient levels and the low cadmium values, litter from intensively managed layers appears as the most suitable for application on agricultural soils. PMID- 16697441 TI - Occurrence and reductions of pharmaceuticals and personal care products and estrogens by municipal wastewater treatment plants in Ontario, Canada. AB - Over the last ten years there have been reports of pharmaceuticals and personal care product (PPCP) residuals in municipal wastewater treatment plant (WWTP) effluents. The principle goal of this study was specifically to expand and in some cases establish a Canadian database for the presence of selected acidic drugs, triclosan, polycyclic musks, and selected estrogens in MWWTP influent and effluent. The impact of treatment configuration (e.g. lagoons, conventional activated sludge (CAS), and CAS followed by media filtration (CAS+filtration)) was also examined. For CAS systems, the most prevalent treatment type, the effect of operating temperature and SRT was evaluated. Selected PPCPs included ten acidic pharmaceuticals (i.e. a group of pharmaceuticals that are extractable at a pH of 2 or less), triclosan, five polycyclic musks and two estrogens. The pharmaceuticals and musks were selected on the basis of levels of use in Canada; reported aquatic toxicity effects; and the ability to analyze for the compounds at low levels. Twelve MWWTPs discharging into the Thames River, the second largest river in southwestern Ontario, were surveyed. The only common characteristic of acidic drugs is their extraction pH as they differ in their intended biological function and chemical structure. Many organics degraded by WWTP processes benefit from warm temperatures and long SRTs so the impact of these variables warranted additional attention. Influent concentrations and reductions for acidic drugs reported by this study were compared to other Canadian studies, when available, and European investigations. The data of this study seems consistent with other reports. Ten acidic drugs were considered by this study. Three were consistently present at non-quantifiable levels (e.g. CLF, FNP and FNF). Additionally, one analyte, SYL, presented results that were so inconsistent that the values were not analysed. The remaining six acidic pharmaceuticals were placed into three categories. IBU and NPX members of the first category had consistently high reductions. At the level of reduction achieved (i.e. median reduction of greater than 93%) and any effect of treatment type or operating characteristics would be subtle and non-discernable given the analytical noise. In the second group are KTP and IND, and definitive comments are difficult to make on the impact of treatment type and operational considerations due to a sparse data set (i.e. many influent values were at non quantifiable concentrations). Median reductions were in the 23% to 44% range. In the last category are GMF and DCF which have median reductions of 66% and -34%, respectively. Several negative reduction values in the data set (i.e. twelve of twenty six sampling events) suggest that DCF may be deconjugated under certain conditions. This warrants further evaluation when analytical methods for measuring human metabolites of DCF are available. For both GMF and DCF, reduction does not appear to be strongly influenced by SRTs up to 15 days, while SRTs over 30 days were associated with more frequent non-quantifiable effluent levels of DCF. This would suggest that better treatment would be provided by lagoons and CAS systems with extended aeration. Preliminary data suggests that temperature does not play a strong role in the reduction of these compounds. Triclosan (TCL) was detected at concentrations of 0.01-4.01 microg/L in influent samples and 0.01 0.324 microg/L in effluent samples. Reduction of TCL ranged from 74% to 98%. Lagoon treatment seems to be the best TCL reduction as it was present in the influent and effluent at quantifiable and non-quantifiable concentrations, respectively, on nine of nine sampling occasions. Influent and reduction values of five polycyclic musks (e.g. ADBI, AHMI, ATII, HHCB, and AHTN) were examined over the course of this study. AHMI was predominantly present at non-quantifiable concentrations. HHCB and AHTN were present at the highest concentrations. A comparison between Canadian values and those of European studies indicate that in general polycyclic musk concentrations in Canadian MWWTP effluents are 5-10 times lower. More extensive European and Canadian databases would be useful in confirming this initial observation. Median reductions for the five remaining musks range between 37% and 65% in CAS systems. CAS+filtration systems would be expected to have higher reductions if musks were bound to the effluent solids. This trend is not apparent but this may be due to the small size of the data set. In lagoon systems, musk reduction for HHCB and AHTN are approximately 98-99%. For ADBI and ATII musk, there are no numerical reduction values as most often the effluent concentration was non-quantifiable. In some instances, both the influent and effluent concentrations were non-quantifiable. The hormones 17-beta-estradiol (E2) and estrone (E1) were detected at concentrations of 0.006 to 0.014 and 0.016 to 0.049 microg/L, respectively. E2 was not detected in any effluent samples (<0.005 microg/L) whereas E1 was detected in effluent samples from CAS treatment plants (median of 0.008 microg/L), and in one sample from lagoons. These data demonstrate that there are detectable levels of PPCPs entering Canadian waterways at trace levels, and that only some of these compounds are being reduced in a significant proportion by municipal wastewater treatment processes. PMID- 16697442 TI - Trees as reservoirs for amphibole fibers in Libby, Montana. AB - Tree bark and core samples were collected from areas surrounding the asbestos contaminated vermiculite mine in Libby, MT. These samples were collected to provide preliminary data in support of a proposed study to determine if trees can serve as reservoirs for amphibole fibers and to determine if there is a potential for exposure to those that harvest contaminated wood in the Libby mine area, specifically during firewood harvesting and commercial logging. Initially, three sets of samples were taken both within and directly outside of the EPA restricted area surrounding the mine site. Based on the results of the initial samples, a follow-up sampling program was conducted both in the town of Libby and directly outside the city limits. Gravimetric reduction of a tree core sample did not indicate the presence of amphibole fibers. However, transmission electron microscopy analysis of bark samples collected near the vermiculite mine yielded substantial amphibole fiber concentrations ranging from 41 million to 530 million fibers/g of bark. In addition, a bark sample collected approximately 7 miles west of the town next to a railroad line had concentrations of 19 million fibers/g. A conversion of these mass-based concentrations to areal concentrations (to reflect surface area contamination) revealed concentrations in excess of 100 million amphibole fibers/cm(2). These preliminary results suggest that trees in the Libby valley and along vermiculite shipping corridors can serve as reservoirs for amphibole fibers, and that a potential for exposure exists for those who harvest contaminated wood. PMID- 16697443 TI - Testicular trauma secondary to less-lethal kinetic energy munitions. AB - Many cases of testicular trauma secondary to munitions have been reported. We report a case of a 37-year-old man who suffered testicular trauma as a result of a less-lethal munition projectile. With the advent, and increased use, of less lethal munitions by the military and law enforcement agencies, more of these new subsets of genitourinary trauma patients who will require care are sure to result. PMID- 16697444 TI - Effects of melatonin on suppression of renal scarring in experimental model of pyelonephritis. AB - OBJECTIVES: To determine the effects of melatonin combined with antibiotic administration on the suppression of renal scarring in an experimental pyelonephritis model. METHODS: The control group underwent a sham operation without infection. In the other groups, treatment began 72 hours after direct bacterial inoculation. In the no-treatment group, rats received daily intraperitoneal injections of saline. In the antibiotic-only group, the rats were treated only with ceftriaxone intramuscularly at a dose of 50 mg/kg once daily for 5 days. In the melatonin-only group, only 20 mg/kg of melatonin once daily was given by intraperitoneal injection for 5 days. In the antibiotic plus melatonin group, melatonin and ceftriaxone were administered at the same dosages and duration as for the single-modality treatment groups. After 6 weeks, the kidneys were removed for malondialdehyde measurements and histopathologic examination (inflammatory response and cicatrization). RESULTS: Melatonin only (134.25 +/- 13.42) and antibiotic plus melatonin treatment (122.62 +/- 8.91) caused a marked reduction in the mean malondialdehyde values compared with no treatment (214.12 +/- 17.77) and antibiotic-only treatment (161.37 +/- 16.03), with no significant difference compared with that of the control group (120.75 +/ 9.83). Histopathologically, in the no-treatment group, the severity of scarring correlated directly with the severity of inflammation (r = 0.93). No significant differences were found in the renal scarring scores in rats receiving no treatment and those treated only with antibiotic or melatonin. In the antibiotic plus melatonin treatment group, the cicatrization score was not statistically different from that of the control group. CONCLUSIONS: When combined with antibiotics, melatonin causes a significant inhibition of malondialdehyde production and neutrophil infiltration caused by acute pyelonephritis in an experimental rat model, and these are responsible for the protective effect of melatonin against renal damage, preventing renal scarring formation. PMID- 16697445 TI - Ureteral obstruction by uterine artery aneurysm in third trimester of pregnancy. AB - Uterine artery aneurysm (UAA) is an uncommon complication of pregnancy. Its diagnosis before postpartum hemorrhage is rare. We report on a patient who presented with symptomatic ureteral obstruction secondary to UAA. The hydronephrosis was managed by ureteral stenting. The UAA was successfully embolized after a planned cesarean section delivery. UAA is a rare and clinically significant cause of hydronephrosis during pregnancy. PMID- 16697446 TI - Adult prone retroperitoneal partial nephrectomy: initial experience. AB - To our knowledge, we provide an initial description of an adult patient undergoing a retroperitoneal partial nephrectomy in the prone position. The prone position was chosen because of the posteromedial location of the patient's right kidney lesion. We describe our surgical methods and the potential pitfalls to this approach. PMID- 16697447 TI - Prognostic significance of neuroendocrine expression in lymph node-positive prostate cancer. AB - OBJECTIVES: To evaluate the expression of chromogranin A, a marker for neuroendocrine (NE) differentiation, in patients with lymph node-positive prostate cancer to determine its prognostic significance. NE cells are involved in cellular growth and differentiation in both normal and pathologic conditions of the prostate. METHODS: We reviewed the data of 140 patients with lymph node positive prostate adenocarcinoma treated with radical prostatectomy and pelvic lymphadenectomy. The median follow-up was 10.9 years (range 0.8 to 19.7). Immunohistochemical staining for chromogranin A was evaluated in areas of benign epithelium, primary prostate cancer, and lymph node metastasis. The association between chromogranin A expression and the clinical and pathologic factors (preoperative serum prostate-specific antigen and prostatectomy Gleason score and stage) and clinical outcomes, including overall and recurrence-free survival, was evaluated. RESULTS: Staining was positive in 86% of benign areas, 61% of primary cancer specimens, and 12% of lymph node deposits. The preoperative serum prostate specific antigen level and pathologic stage and grade of the primary tumor did not show any statistically significant correlation with NE staining in any of the areas. Only NE expression in the primary tumor was associated with clinical recurrence, with a 10-year recurrence-free survival rate for those with less than 5% staining of 67% compared with 35% for those with 5% staining or greater (P = 0.03). Furthermore, after adjusting for age, greater NE expression in the primary tumor (relative risk 2.15, P = 0.02) and lymph node deposit (relative risk 2.03, P = 0.03) was associated with poorer overall survival. CONCLUSIONS: NE expression in the primary tumor and lymph node metastasis of patients with node-positive prostate cancer may provide additional prognostic stratification. PMID- 16697448 TI - Complete penoscrotal transposition. PMID- 16697449 TI - HIV-associated primary non-Hodgkin's lymphoma of ovary: a case report. AB - BACKGROUND: Although there have been few case series of primary non-Hodgkin's lymphoma (NHL) of ovary, no patient with AIDS-related primary ovarian lymphoma has been described in the literature. CASE: We report a case of human immunodeficiency virus infected female with primary ovarian NHL for which she could not receive standard NHL combination chemotherapy and standard HIV highly active antiretroviral therapy (HAART) and succumbed to death within 7 months after diagnosis. CONCLUSIONS: Primary ovarian lymphoma is rare as compared with other ovarian tumors and has similar presentation to other common ovarian malignancies. The patients from developing countries cannot afford standard NHL combination chemotherapy and HAART as these drugs are expensive and are not provided freely to all patients, in state government run public hospitals. PMID- 16697450 TI - Pathological characteristics of cervical adenocarcinoma in a multi-center US based study. AB - OBJECTIVE: Difficulties in detecting cervical adenocarcinoma early are well known. We report a detailed pathology review of cervical adenocarcinoma subtypes, comparing growth patterns and appearance of non-neoplastic epithelium to inform possible clues for disease progression and early detection. METHODS: This analysis includes 154 women aged 18-69 years and diagnosed with incident in situ or invasive adenocarcinoma (AC), adenosquamous (AS), or other rare cervical glandular tumors from 1992-1996 in six U.S. medical centers. A pathology review panel evaluated histological features from original diagnostic slides. RESULTS: Higher tumor grade (P < 0.001) and vascular invasion (P = 0.002) were more common in AS compared to AC. Adenocarcinoma in situ (AIS) was also more common among AC than AS (P = 0.002). Among AC with cervical intraepithelial carcinoma (CIN), AIS and cribriform patterns were more common than AC without CIN (P = 0.01). Further, non-endometrioid AC had higher tumor grade (P = 0.01) and stromal responses (P = 0.02) than endometrioid AC. Finally, although microglandular hyperplasia is historically thought to be related to oral contraceptive (OC) use, our data do not support this notion. CONCLUSION(S): AS appears to be either diagnosed later or histologically more aggressive than AC, and among AC subtypes, there are distinct histologic characteristics. Further research is needed to identify precursor lesions for early detection of AC and particularly for AS where AIS may not be a common precursor. PMID- 16697451 TI - Sustained response to bevacizumab in refractory well-differentiated ovarian neoplasms. AB - BACKGROUND: Bevacizumab has demonstrated activity against a variety of solid tumors, including ovarian carcinoma. However, there have not been reproducible prognostic features associated with its activity. CASES: One patient each with recurrent, refractory well-differentiated serous-endometrioid ovarian carcinoma, micropapillary serous carcinoma of the ovary, and primary peritoneal micropapillary serous carcinoma were treated with single agent bevacizumab (15 mg/kg [DOSAGE ERROR CORRECTED] intravenously every 3 weeks). All three have had dramatic sustained responses of 15, 15, and 22 months' duration. CONCLUSION: Bevacizumab may have significant activity against well-differentiated ovarian carcinoma and micropapillary serous carcinomas of the ovary or peritoneum. Since these tumors are generally indolent and not responsive to adjuvant therapy, further investigation is warranted. PMID- 16697452 TI - Influencing healthful food choices in school and home environments: results from the TEENS study. AB - BACKGROUND: The purpose of this research is to examine the effects of an intervention designed to increase the availability of fruits, vegetables and lower fat foods in homes and schools. This research is part of the TEENS study, a school-based intervention study. METHODS: Sixteen schools in Minnesota were recruited to be in the study, and approximately 3600 middle school students in the eight intervention schools were exposed to a multi-component intervention. The TEENS intervention included classroom-based curricula, family newsletters, and changes in the school food environment including increasing more healthful options on a la carte and on the school lunch line. In addition to student-level outcomes, changes in availability of fruits, vegetables, and lower fat snacks in home and school environments were evaluated. The TEENS study was conducted from 1997 to 2000. RESULTS: Parents of students in intervention schools reported making healthier choices when grocery shopping as compared to parents of students in control schools (P = 0.01). No intervention effects were evident from a home food inventory. Compared to control schools, intervention schools offered (P = 0.04) and sold (P = 0.07) a higher proportion of healthier foods on a la carte, but no effects were seen for fruit and vegetables sales as part of the regular meal pattern lunch. CONCLUSION: Our results show mixed results for positively influencing adolescents' school and home environments. PMID- 16697454 TI - Adhesive systems and secondary caries formation: Assessment of dentin bond strength, caries lesions depth and fluoride release. AB - OBJECTIVES: The present study evaluated the microtensile bond strength and caries formation on adhesive/dentin interfaces before and after dynamic chemical formation of secondary caries. METHODS: Restorations were prepared on the dentin surface of 80 bovine incisors using four adhesive systems: two fluoride-free (Single Bond and Clearfil SE Bond) and two fluoride containing (Optibond Solo Plus and Clearfil Protect Bond). The restored teeth were then sectioned into multiple slabs that were further trimmed at the bonded interface to a cross sectional area of 1 mm2. Half of the slabs were subjected to secondary caries formation using a pH cycling model (treated groups); while the other half was used as the control group (no pH cycling). The specimens designated for bond strength evaluation were subjected to microtensile bond strength test (muTBS). Caries lesions formation was assesses by polarized light microscopy at different depths from the adhesive-dentin bonded interface. The fluoride ion concentration was evaluated using the de/remineralization solutions (De/Re). RESULTS: No differences in muTBS were observed among the adhesive systems in both the control and treated conditions. Secondary caries significantly reduced the values of muTBS for all adhesives (p<0.05). Optibond Solo Plus presented the lowest caries formation at 5 microm depth. Fluoride concentrations present in the De/Re were less than 0.03 ppm, regardless of the adhesive system tested. SIGNIFICANCE: Bond strength values significantly decreased after in vitro secondary caries formation. Fluoride present in adhesive systems is not capable of inhibiting secondary caries or maintaining bond strength values following caries formation. PMID- 16697453 TI - Uncoupled leftward asymmetries for planum morphology and functional language processing. AB - Explanations for left hemisphere language laterality have often focused on hemispheric structural asymmetry of the planum temporale. We examined the association between an index of language laterality and brain morphology in 99 normal adults whose degree of laterality was established using a functional MRI single-word comprehension task. The index of language laterality was derived from the difference in volume of activation between the left and right hemispheres. Planum temporale and brain volume measures were made using structural MRI scans, blind to the functional data. Although both planum temporale asymmetry (t(1,99) = 6.86, p < .001) and language laterality (t(1,99) = 15.26, p < .001) were significantly left hemisphere biased, there was not a significant association between these variables (r(99) = .01,ns). Brain volume, a control variable for the planum temporale analyses, was related to language laterality in a multiple regression (beta = -.30, t = -2.25, p < .05). Individuals with small brains were more likely to demonstrate strong left hemisphere language laterality. These results suggest that language laterality is a multidimensional construct with complex neurological origins. PMID- 16697455 TI - Use of fish in vitro hepatocyte assays to detect multi-endpoint toxicity in Slovenian river sediments. AB - There is an increasing demand for rapid, sensitive and robust methods for toxicity testing of single chemicals, complex mixtures and environmental samples. The objective of this work was to validate and use a primary culture of rainbow trout (Oncorhynchus mykiss) hepatocytes as a multi-endpoint in vitro bioassay for toxicity characterisation of river sediments from four areas of the Sava and Krupa Rivers (Slovenia). The endpoints were chosen to encompass acute toxicity (cytotoxicity) as well as sub-lethal biomarker and effect endpoints such as metabolic inhibition, DNA damage (Fast Micromethod), endocrine disruption (estrogenicity), and 7-ethoxyresorufin O-deethylase (EROD) activity. Results from these studies show that the primary hepatocyte culture was able to successfully detect effects of single model chemicals in all endpoints analysed. Furthermore, the bioassays were also able to discriminate between contaminated and less contaminated sediments for a number of endpoints such as cytotoxicity, metabolic inhibition and induction of EROD activity, although no increase in DNA damage and estrogenicity was observed above background at any site. The present study shows that primary fish hepatocytes may be used to determine multiple mechanisms of toxic action and that a holistic assessment of effects may improve our understanding of cellular toxicity of complex mixtures such as sediments. PMID- 16697456 TI - Developmental differences in elimination of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) during Xenopus laevis development. AB - Although 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) is a potent developmental toxicant in most vertebrates, several frog species are insensitive to TCDD, especially during early life stages. Previous experiments with ranid frogs suggest that TCDD insensitivity results largely from rapid elimination. Recent studies in the African clawed frog (Xenopus laevis; family Pipidae) link low TCDD toxicity with the low binding affinity of aryl hydrocarbon receptors, which mediate the toxic effects of dioxin-like compounds. The present study sought to examine TCDD elimination in X. laevis embryos and tadpoles, enabling an integrated assessment of the relative roles of TCDD elimination and AHR-related mechanisms in TCDD insensitivity within a single frog species. Using tadpoles (stage 52-55; approximately 1 month old) exposed to [3H]TCDD, we observed that TCDD has a relatively short half life of 102.6 h, consistent with other frogs and much faster than reported clearance rates in developing fish. In contrast, TCDD elimination is much slower during early development. Embryos exposed during primary organogenesis (from stage 31-41, beginning approximately 36 h after fertilization) exhibited little TCDD elimination during the subsequent 96 h. Enhanced TCDD clearance in later developmental stages may follow the appearance of a functional digestive tract and the onset of feeding. These results suggest that rapid elimination is unlikely to contribute mechanistically to TCDD insensitivity during development of the cardiovascular system, which is significantly perturbed by TCDD in fish embryos. PMID- 16697457 TI - Organ-specific patterns of P450arom gene isoforms are modulated by p,p'-DDE in adult male European common frog, Rana temporaria. AB - The organ-specific gene expression patterns of P450arom isoforms have been studied in European common frog, Rana temporaria after exposure to DDE, using real-time PCR. Four groups of frogs were subcutaneously injected with DDE at 0.01, 0.1, 1 and 10 mg/kg body weight and one group, serving as the control was injected with pure corn oil. Brain, kidney, testis and liver P450aromA and P450aromB gene expressions were evaluated at day 14 after exposure. P450aromB data show that 0.1 and 10 mg DDE/kg doses caused 76% and 63% (testis) and 79% and 80% reductions, respectively, of mRNA levels. Brain P450aromB mRNA decreased 10% and 34%, respectively, after exposure to 0.01 and 0.1 mg DDE/kg. Thereafter, a 185% and 177% induction response of brain P450aromB was observed in the groups treated with 1 and 10 mg DDE/kg, respectively. In the kidney, 0.01, 0.1, 1 and 10 mg DDE/kg induced a 516%, 178%, 466% and 247% induction of P450aronB mRNA, respectively. P450aromA expression was not quantified in any of the organs. The relative abundance of P450aromB gene expression in different organs is in the order: kidney > brain > liver > testis. The present data suggest potential detrimental effect of organochlorine pesticides (OCs) and their metabolites on the European frog steroidogenic pathways. Given the high persistency in the environment and continued use in developing countries coupled with the tendency for global atmospheric transport, OCs and their metabolites such as DDE will remain a focus of concern both for scientific and societal reasons. PMID- 16697458 TI - In vivo and in vitro CYP1B mRNA expression in channel catfish. AB - Our goal was to study the induction of CYP1B mRNA expression in channel catfish (Ictalurus punctatus). CYP1B belongs to the cytochrome P450 superfamily of genes, is involved in the oxidation of endogenous and exogenous compounds, and could potentially be a useful biomarker in fish for exposure to AhR ligands. The full length catfish CYP1B cDNA is 2417 nt to the polyA tail and encodes a putative protein of 536 amino acids. It has 67% amino acid similarity to carp and zebrafish CYP1B and 68% similarity to carp CYP1B2. Male channel catfish were collected from three Mississippi Delta sites: Lake Roebuck, Itta Bena; Bee Lake, Thornton; and Sunflower River, Indianola. Total RNA was isolated from wild-caught catfish gill, blood, gonad and liver tissues. Quantitative real-time reverse transcriptase PCR was used to determine relative induction of CYP1B in wild catfish compared to laboratory control and BaP-exposed catfish (20mg/kg i.p. after 4 days). BaP exposure significantly induced CYP1B message in blood, gonad, and liver of laboratory catfish. In these same tissues of wild catfish from sites with relatively low sediment contaminants, CYP1B message was not statistically increased relative to laboratory control catfish. CYP1B transcript abundance was higher in gills compared to other tissues in both laboratory and wild catfish. When primary cultured gill cells were treated with increasing concentrations of BaP, TCDD, and PCBs 77, 126 and 169, CYP1B mRNA was induced more than 10-fold while PCB153 and 4,4'DDT did not cause significant CYP1B induction. Our results suggest that catfish CYP1B is induced by the classic AhR ligands. PMID- 16697459 TI - Effects of algal extracts (Polysiphonia fucoides) on rainbow trout (Oncorhynchus mykiss): a biomarker approach. AB - The genotoxicity of algal extracts (Polysiphonia fucoides) was investigated in erythrocytes of rainbow trout (Oncorhynchus mykiss). Trout were exposed to 0.5% of the algal extract for 7 days. Comet assay (alkaline and neutral versions) and Micronucleus test were used to assess DNA damage, and Diffusion Assay to detect apoptotic cells. EROD activities and oxidative stress parameters in rainbow trout liver were also measured. A significant induction of DNA single strand breaks comparable to the ones induced by the in vivo exposure to 20 mg/kg B[a]P was observed at the end of the treatment, while increases of double strand breaks and apoptotic cells were not observed. The absence of activation of antioxidant responses seems to underline a mechanism of action of the genotoxic algal extract which does not involve oxidative stress. PMID- 16697460 TI - The environmental estrogen, 4-nonylphenol modulates brain estrogen-receptor- and aromatase (CYP19) isoforms gene expression patterns in Atlantic salmon (Salmo salar). AB - The modulations of reproductive biomarker responses, such as vitellogenin (Vtg) and zona radiata protein (Zr-protein) by xenoestrogens are mediated through the estrogen receptors (ERs). The ERalpha and ERbeta belong to the nuclear receptor (NR) superfamily of ligand activated transcription factors that modulate specific gene expression. By interacting with estrogen and estrogen like molecules (including estrogen mimics), the ERs exert a diverse array of biological effects. In addition to their effect as direct agonist to the ERs, the endocrine disrupting effects of environmental chemicals are sometimes interpreted as interference with steroidogenesis through the aromatase enzyme (CYP19 or P450arom). The aim of this study was to investigate the effects of nonylphenol on the expression patterns of ER and aromatase (P450arom) isoform genes in the brain as sensitive molecular biomarker of response to endocrine disrupting chemicals. Immature Atlantic salmon were exposed to continuous nominal waterborne nonylphenol at 15 microg/L. Brain samples were collected after 7 days exposure. A quantitative RT-PCR method was used to analyze mRNA expression patterns. Nonylphenol caused a significant induction of P450arom and ER isoform patterns in the brain and represent quantitative gene expression biomarkers of exposure to endocrine disrupting chemicals in the environment. PMID- 16697461 TI - Effluents from oil production activities contain chemicals that interfere with normal function of intra- and extra-cellular estrogen binding proteins. AB - Some environmental pollutants have the ability to alter the endocrine function in fish through interaction with the estrogen receptor (ER). Many of these chemicals are also able to interfere with the endocrine system through other mechanisms of action, however. The plasma sex steroid-binding protein (SBP), which is involved in regulating circulating levels of endogenous sex steroids, has recently been proposed to contribute to pollutant induced disruption of endocrine homeostasis. The objective of the present work was to determine whether industrial effluents contain chemicals that are able to modulate the endocrine system through interference with the function of the ER and SBP using in vitro biological assays (bioassays) from rainbow trout. The results show that solid phase extracts of process water (produced water) from an oil production facility in the North Sea and a land-based oil refinery contain chemicals that are able to induce estrogenic effects as well as displace natural sex steroid 17beta-estradiol from the SBP. The bioactive chemicals were found to be partly resistant to biological degradation, but the identity of the chemicals was not determined. The alkylphenol 4-tert-butylphenol, which is known to occur in effluents from various oil production facilities, was found to be estrogenic and displace 17beta estradiol from the SBP and may thus contribute to the observed endocrine disrupting activity. PMID- 16697462 TI - How does the legal system respond when children with learning difficulties are victimized? AB - OBJECTIVE: To understand how the Swedish legal system perceives and handles mentally handicapped children who may have been victimized. METHOD: Twenty-two judicial districts in Sweden provided complete files on 39 District Court cases (including the Appeals Court files on 17 of these cases) involving children with learning difficulties or other handicaps as alleged victims of abuse, threat and neglect. The children (25 girls and 14 boys) averaged 11.8 years of age when first allegedly victimized. Sexual abuse was the most frequently alleged crime (33 cases). Court transcripts, court files and expert assessments of the alleged victims' handicaps and their possible consequences were examined to elucidate the ways in which courts evaluated the credibility of the alleged victims. RESULTS: The children's reports of their victimization were expected to have the characteristics emphasized by proponents of Statement Reality Analysis (SRA) and Criterion Based Content Analysis (CBCA) in order to be deemed credible. Expert reports were seldom available or adequate. Because many reports were poorly written or prepared by experts who lacked the necessary skills, courts were left to rely on their own assumptions and knowledge when evaluating children's capacities and credibility. CONCLUSIONS: Children with learning difficulties or other handicaps were expected to provide the same sort of reports as other children. To minimize the risk that judgments may be based on inaccurate assumptions courts need to require more thorough assessments of children's limitations and their implications. Assessments by competent mental health professionals could inform and strengthen legal decision-making. A standardized procedure that included psycho-diagnostic instruments would allow courts to understand better the abilities, capacities, and behavior of specific handicapped children. PMID- 16697464 TI - Civilising missions, natural history and British industry: Livingstone in the Zambezi. AB - High hopes for opening new markets, locating natural resources and the 'advance of civilisation' were built into the plans for Victorian expeditions to Africa. When an expedition failed to deliver on these goals, criticisms of individual explorers and the entire 'civilising mission' project could result. The Zambesi Expedition, led by David Livingstone, received just such criticism when politics and geography obstructed the realisation of its aims. The needs of a growing industrial sector contributed to the organisation of the Expedition but also set the criteria by which its success or failure would be judged. PMID- 16697465 TI - A human monoclonal IgG1 potently neutralizing the pro-inflammatory cytokine GM CSF. AB - The pro-inflammatory cytokine GM-CSF is aberrantly produced in many autoimmune and chronic inflammatory human diseases. GM-CSF neutralization by antibodies has been shown to have a profound therapeutic effect in animal models of rheumatoid arthritis, inflammatory lung diseases, psoriasis and multiple sclerosis. Moreover, the absence of GM-CSF in null mutant mice ameliorates or prevents certain of these diseases. Here we describe the biophysical and biological properties of a human anti-GM-CSF IgG1 antibody designated MT203, which was derived by phage display guided selection. MT203 bound with picomolar affinity to an epitope on human and macaque GM-CSF involved in high-affinity receptor interaction. As a consequence, the antibody potently prevented both GM-CSF induced proliferation of TF-1 cells with a sub-nanomolar IC50 value and the production of the chemokine IL-8 by U937 cells. MT203 neutralized equally well recombinant (r) human (h) GM-CSF from Escherichia coli and yeast, and also normally glycosylated GM-CSF secreted by human lung epithelial cells in response to IL-1beta stimulation. Furthermore, MT203 significantly reduced both survival and activation of peripheral human eosinophils as may be required for effective treatment of inflammatory lung diseases. The antibody did not show a detectable loss of neutralizing activity after 5 days in human serum at 37 degrees C. Based on its favorable properties, MT203 has been selected for development as a novel anti-inflammatory human monoclonal antibody with therapeutic potential in a multitude of human autoimmune and inflammatory diseases. PMID- 16697466 TI - A new format of electrodes for the electrochemical reduction of cytochromes P450. AB - New approach to the electrochemical reduction of cytochromes P450 (P450s, CYPs) at electrodes chemically modified with appropriate substrates for P450s ("reverse" electrodes) was proposed. The method is based on the analysis of cyclic voltammograms, square-wave voltammograms and amperograms with subsequent determination of electrochemical characteristics such as catalytic current and redox potential. The sensitivity of proposed method is 0.2-1 nmol P450/electrode. The changes of maximal current and of redox potentials in square-wave voltammograms as well as the changes of catalytic current in amperometric experiments proved to be informative and reliable. Planar regime of screen printed electrodes (strip-type sensors) enabled to utilise 20-60 microl of electrolyte volume. The enzyme-substrate pairs P450 2B4/benzphetamine and P450scc/cholesterol were investigated. Electrochemical parameters of electrodes with unspecific P450 substrates differed considerably from electrodes with appropriate substrates. PMID- 16697467 TI - Chiral separation of T3 enantiomers using stereoselective antibodies as a selector in micro-HPLC. AB - This work deals with the application of stereoselective antibodies against L-T3 as a tailor-made chiral selector in micro-HPLC. The separations were performed in microbore columns using commercially available anti-L-T3 antibodies chemically bonded to 5 microm silica gel. The enantiomers of T3 were baseline separated under mild continuous isocratic elution conditions using 10 mM phosphate buffer, pH 7.4. The D-enantiomer eluted with the void volume, while the L-enantiomer was retained by the antibody phase and eluted second. An indirect competitive and non competitive enzyme linked immunosorbent assay (ELISA) was used for testing the stereoselectivity of anti-L-T3 antibodies. PMID- 16697468 TI - The Brief-HAPPI: a questionnaire to assess cognitions that distinguish between individuals with a diagnosis of bipolar disorder and non-clinical controls. AB - BACKGROUND: Despite developments in cognitive behavioural therapy for bipolar disorder, little is known about the specific dysfunctional beliefs that may predispose individuals to mania. Therefore a measure was specifically designed for this purpose based on a cognitive approach. The measure is called the Hypomanic Attitudes and Positive Predictions Inventory (HAPPI). METHOD: Fifty-six individuals with a diagnosis of bipolar disorder and 39 matched healthy controls completed a brief version of the HAPPI (Brief-HAPPI), the Internal State Scale, and the Hypomanic Interpretations Questionnaire (HIQ). RESULTS: The bipolar group scored higher on the overall Brief-HAPPI scale, higher on the forward items of the Brief-HAPPI, lower on the reverse items and no different on the filler items. The group differences in overall score remained when controlling for current self reported symptoms of mania and depression, and recent history of hypomanic symptoms, yet within the bipolar group, Brief-HAPPI score was positively correlated with the level of symptoms. The Brief-HAPPI showed a sizeable correlation with the HIQ, yet each scale showed a significant and independent association with bipolar disorder. CONCLUSION: The Brief-HAPPI assesses dysfunctional beliefs that are associated with a diagnosis of bipolar disorder. PMID- 16697469 TI - Anorexia nervosa with excessive exercise: a phenotype with close links to obsessive-compulsive disorder. AB - Anorexia nervosa (AN) and obsessive-compulsive disorder (OCD) are highly comorbid, and appear to share a common neurophysiological dysfunction that contributes to the obsessional thoughts and compulsive behaviours seen in both disorders. Obsessive-compulsive personality (OCP) traits are also important risk factors for AN. Since excessive exercise has also been associated with greater obsessionality, we hypothesised that AN patients with a hyperactive behavioural profile represent a phenotype more closely linked to OCD than their non exercising counterparts. We examined prospectively 50 female AN-Restrictor patients whom we classified as "excessive" or "non-excessive" based on their exercise status i) at admission and ii) over the lifetime of their illness. Validated measures of OCD symptoms and OCP traits were obtained at admission and after refeeding at discharge. On both classification methods, excessive exercisers had greater OCD symptoms and OCP traits than the non-excessive group, but did not differ on body mass index. OCD symptoms, but not OCP traits, decreased between admission and discharge. Findings support our prediction that AN patients with excessive physical activity constitute a subtype of the disorder with strong links to OCD. Indeed, this phenotype may be a culture-bound variant of OCD. PMID- 16697470 TI - Social anxiety and obsessive-compulsive spectra: validation of the SHY-SR and the OBS-SR among the Spanish population. AB - The study focuses on the adaptation into Spanish and on the validation of the Social Phobia Spectrum Self-Report (SHY-SR) and the Obsessive-Compulsive Spectrum Self-Report (OBS-SR). The questionnaires were designed to measure a broad range of subtle and atypical features related to social anxiety and obsessive compulsive phenomenology, respectively. Sixty-two outpatients who met DSM-IV criteria for social phobia (SP, n = 20), obsessive-compulsive disorder (OCD, n = 22) and major depression (MD, n = 20), and 25 non-clinical subjects participated. The spectra questionnaires were administered along with the Liebowitz Social Anxiety Scale and the Maudsley Obsessional Compulsive Inventory. The instruments proved to have satisfactory internal consistency and test-retest reliability. Convergent validity with other instruments was excellent for the SHY-SR and moderate for the OBS-SR. Both questionnaires were able to detect differences between patients with the disorder of interest (SP in the case of the SHY-SR scores and OCD in the case of the OBS-SR scores) and either normal controls or patients with MD. Receiver-Operating Characteristic Curve analyses were conducted to determine cut-off values in the Spanish versions of the questionnaires denoting the presence of significant SP and OCD symptomatology. Are the questionnaires available on the website? PMID- 16697471 TI - Reduced platelet [3H]paroxetine binding in anorexia nervosa: relationship to eating symptoms and personality pathology. AB - Alterations in serotonin function have been implicated in both anorexia and bulimia nervosa, and previous studies suggest associations between serotonin function and variations in pathological personality traits. Women meeting DSM-IV criteria for anorexia nervosa (AN, 16 with the restricting subtype and 14 with the binge-purge subtype) and 49 healthy control women (CW) provided blood samples for analyses of platelet [(3)H]paroxetine binding. Participants also filled out questionnaires tapping eating disorder symptoms, depression, and personality pathology. Compared with CW, women with restricting and binge-purge AN had significantly lower levels of paroxetine binding (respectively: 1012 + 487 vs. 560 + 253 vs. 618 + 217 fmol/mg protein). Simple correlation analyses showed that, within AN but not within controls, paroxetine binding was inversely related to dieting preoccupations, affective instability, anxiousness, compulsivity, restricted expression and social avoidance but independent of age, body mass index, depression, and other eating symptoms. Findings suggest that reduced peripheral serotonin transporter density in AN relates to increased dieting preoccupations, affective instability and anxiousness-fearfulness. PMID- 16697473 TI - Angiogenesis: from plants to blood vessels. AB - Angiogenesis is a major pathological component of diseases such as cancer and coronary heart disease. Although major advances have been made and encouraging clinical results obtained, safer and more effective approaches are required. The identification of new drugs from plants has a long and successful history, and certain proangiogenic and antiangiogenic plant components have been used in traditional Chinese medicine (TCM) for thousands of years. Similar to Western combination therapy, TCM uses mixtures of plant extracts, termed fufang, to maximize efficacy and minimize adverse effects or toxicity. More evidence-based research and chemical optimization of these compounds could further enhance the effectiveness of these plant-based medicines in angiotherapy. PMID- 16697474 TI - Rats assess costs and benefits according to an internal standard. AB - Variation in effort to obtain rewards is a fact of mammalian everyday life. In this study, we assess how rats scale variable costs and benefits. Different groups of rats were trained in a T-maze to discriminate a high (three or five sugar pellets) from a low reward (one sugar pellet) arm. Subsequently barriers were introduced at the high and low reward side such that the overall long-term pay-off of the high reward arm finally became lower than that of the low reward arm. The data show that under different regimes of costs (climbing barriers) and benefits (number of rewards) of the two arms rats appear to shift their behaviour towards the better side according to a constant relative cost-benefit ratio between the arms. Such a ratio allows them to deal with variation in the (physical appearance of) costs and benefits and choose the best long-term option. PMID- 16697475 TI - Unilateral hemispherectomy at adulthood asymmetrically affects immobile behavior of male Swiss mice. AB - In order to test the hypothesis that behavioral coping with stressful situations is asymmetrically modulated by the hemispheres, we used the unilateral hemispherectomy procedure to assess the relative importance of each hemisphere in the determination of the immobility time during the forced swimming test. Under anesthesia, adult Swiss male mice were submitted to unilateral (right or left) hemispherectomy or sham surgery. Fifteen days after surgery, the immobile and turning behaviors of each mouse were measured during a 5-min forced swimming testing session. In general, while turning activity decreased significantly as the test progressed, an increase in immobility was observed. The unilateral hemispherectomy asymmetrically affected the immobility time in the forced swimming test. Particularly, the increase in immobility time of right hemispherectomized mice was greater than that observed for sham-operated ones. In contrast, there were no differences in turning activity between the groups. The higher immobility time in males that had their right hemisphere removed supports the hypothesis that the two hemispheres contribute differentially to the behavioral response to stress. PMID- 16697477 TI - Homogeneous versus heterogeneous probes for microbial ecological microarrays. AB - Microbial ecological microarrays have been developed for investigating the composition and functions of microorganism communities in environmental niches. These arrays include microbial identification microarrays, which use oligonucleotides, gene fragments or microbial genomes as probes. In this article, the advantages and disadvantages of each type of probe are reviewed. Oligonucleotide probes are currently useful for probing uncultivated bacteria that are not amenable to gene fragment probing, whereas the functional gene fragments amplified randomly from microbial genomes require phylogenetic and hierarchical categorization before use as microbial identification probes, despite their high resolution for both specificity and sensitivity. Until more bacteria are sequenced and gene fragment probes are thoroughly validated, heterogeneous bacterial genome probes will provide a simple, sensitive and quantitative tool for exploring the ecosystem structure. PMID- 16697476 TI - Global genetic diversity and evolution of var genes associated with placental and severe childhood malaria. AB - In Plasmodium falciparum, var genes encode adhesive proteins that are transported to the surface of infected erythrocytes and act as major virulence determinants for infected erythrocyte binding and immune evasion. Var genes are highly diverse and can be classified into five major groups (UpsA, B, C, D, and E). Previous serological studies have suggested that the UpsA var group may contain common antigenic types that have important roles in severe childhood malaria. Here, our analysis found that UpsA vars are highly diverse between 22 world-wide parasite isolates, although they could be grouped into two broad clusters that may be separately recombining. By comparison, orthologs of the UpsA-linked Type 3 var and UpsE-linked var2csa were detected in nearly all parasite isolates, and a var2csa ortholog was also present in a chimpanzee malaria P. reichenowi that diverged from P. falciparum approximately 5-7 million years ago. Although the specific function of Type 3 var genes is unknown, var2csa is a leading candidate for a pregnancy associated malaria vaccine. Compared to typical var genes, var2csa is unusually conserved but still had only 54-94% amino acid identity in extracellular binding regions. However, var2csa alleles have extensive gene mosaicism within polymorphic blocks that are shared between world-wide parasite isolates and recognizable in P. rechenowi suggesting a high rate of self-self recombination and an ancient and globally-related pool of var2csa polymorphism. These studies aid our understanding of the evolutionary mechanisms that shape var diversity and will be important to the development of vaccines against pregnancy associated malaria and severe malaria. PMID- 16697478 TI - Natural drug extracts for a nutritive-tonic drink, promotes the induction of long term potentiation in rat hippocampal dentate gyrus in vivo. AB - We have shown previously that oral administration of a nutritive-tonic drink (NTD) improves scopolamine-induced memory impairment in the passive avoidance task and Morris water-maze in mice and that this action is attributable to the natural drug extracts, rather than synthetic drugs such as taurine and caffeine, in the NTD. In order to investigate the mechanism underlying the antiamnesic effects of the natural drug extracts, the effects of the extracts on the induction of long-term potentiation (LTP) was investigated in the dentate gyrus (DG) of normal and scopolamine-treated rats. Oral administration of natural drug extracts enhanced the induction of population spike amplitude induced by weak tetanic stimulation (30 pulses at 60 Hz). Scopolamine (0.3 mg/kg, i.p.) completely inhibited the induction of LTP induced by both weak and strong tetanic stimulation (100 pulses at 100 Hz). Natural drug extracts enhanced partially but significantly the induction of LTP by strong tetanus, but had a very weak effect on that induced by weak tetanus. These results suggest that LTP induced by strong tetanus is sensitive to natural drug extracts, and that the antiamnesic effect of the NTD is at least partly attributable to the LTP-improving effect of the natural drug extracts in the DG. PMID- 16697479 TI - Development of a quantitative real-time PCR assay for the detection of Aspergillus carbonarius in grapes. AB - Aspergillus carbonarius is the main species responsible for the production of ochratoxin A (OTA) in wine grapes. To monitor and quantify A. carbonarious in grapes, a quantitative real-time PCR assay was developed as a possible tool for predicting the potential ochratoxigenic risk. DNA extraction from grape berries was performed by using conventional extraction and clean up through EZNA Hi-bond spin columns. A TaqMan probe was used to quantify A. carbonarius genomic DNA in grape berries samples. An exogenous internal positive control was used to overcome DNA recovery losses due to matrix inhibition. The quantification of fungal genomic DNA in naturally contaminated grape was performed using the TaqMan signal versus spectrophotometrically measured DNA quantities (Log10) calibration curve with a linearity range from 50 to 5 x 10(-4) ng of DNA. A positive correlation (R2=0.92) was found between A. carbonarious DNA content and OTA concentration in naturally contaminated grape samples. This is the first application of TaqMan real-time PCR for identifying and quantifying A. carbonarius genomic DNA occurring in grapes. The rapid DNA extraction method for grapes, together with the commercial availability of reagents and instrumentation, allows to perform a remarkable number of reproducible assays (96 well format) in less than 4 h. PMID- 16697480 TI - Mechanism of action, development and clinical experience of recombinant FVIIa. AB - Recombinant FVIIa has been developed for treatment of bleedings in hemophilia patients with inhibitors, and has been found to induce hemostasis even during major surgery such as major orthopedic surgery. Recombinant FVIIa is being produced in BHK cell cultures and has been shown to be very similar to plasma derived FVIIa. The use of rFVIIa in hemophilia treatment is a new concept of treatment and is based on the low affinity binding of FVIIa to the surface of thrombin activated platelets demonstrated in a cell-based in vitro model. By the administration of pharmacological doses of exogenous rFVIIa the thrombin generation on the platelet surface at the site of injury is enhanced independently of the presence of FVIII/FIX. As a result of the increased and rapid thrombin formation, a tight fibrin hemostatic plug is being formed. A tight fibrin structure has been found to be more resistant to fibrinolytic degradation thereby helping to maintain hemostasis. The general mechanism of action of pharmacological doses of rFVIIa shown to induce hemostasis not only in hemophilia, but also in patients with platelet defects, and with profuse bleedings triggered by extensive surgery or trauma, may very well be the capacity of generating a tight fibrin hemostatic plug through the increased thrombin generation. Such a fibrin plug will help to resist the overwhelming mostly local release of fibrinolytic activity triggered by the vast tissue damage occurring in extensive trauma. A release of fibrinlytic activity locally has also been demonstrated to occur in the gastrointestinal tract as well as during profuse postpartum bleedings. Pharmacological doses of rFVIIa have in fact, also been shown to induce hemostasis in such cases. PMID- 16697481 TI - Discovery and investigation of a new, second triose phosphate isomerase in Klebsiella pneumoniae. AB - In this study, a tpi1 gene encoding for the enzyme triose phosphate isomerase in Klebsiella pneumoniae DSM2026 was knocked out in an effort to metabolically engineer this strain as a model system for the production of 1,3-propanediol. Investigations of the tpi1 knockout mutant led to the discovery of a second tpi gene (tpi2) in this organism. The new tpi2 gene was cloned and sequenced. The coding region of the tpi2 gene contains 795bp (base pairs) and the deduced protein consists of 265 amino acids. Sequence comparison of TPI2 proteins in different organisms revealed the presence of a highly conserved signature A-Y-E-P V-W-A-I-G-[EDVS]-[GKNASH], which is nearly the same as the reported TPI consensus signature. The tpi1 gene of K. pneumoniae DSM2026 shows a high sequence similarity to that of E. coli, whereas, the tpi2 gene resembles more its relatives in the alpha-proteobacteria, suggesting that they evolve from different ancestors. The overexpression of the tpi2 gene restores the growth deficiency of tpi1 knockout mutant on the minimal medium containing glucose or glycerol. Furthermore, the catalytic activity of this new triose phosphate isomerase was confirmed in both tpi1 knockout mutant and tpi2 over-expressing strain by enzyme assays. For the first time, the co-existence of two tpi genes in an enteric bacterium is experimentally confirmed. PMID- 16697482 TI - Covalent immobilization of triacylglycerol lipase onto functionalized novel mesoporous silica supports. AB - A novel mesoporous silica material was synthesized via a silicate salt route in the presence of polyvinyl alcohol as the structure-directing agent under acidic conditions. The material was functionalized and employed as the supports (LPS-1 and LPS-2) for immobilizing triacylglycerol lipase from porcine pancreas (PPL). Not only they had a good thermal stability and reusability but also the activity recovery of LPS-1 and LPS-2 reached to 69% and 76%, respectively. The optimal pH and temperature region of the LPS supports immobilized PPL for hydrolysis of olive oil were at 8.0 and 55-60 degrees C. Kinetic parameters such as maximum velocity (V(max)) and the Michaelis constant (K(m)) were determined for the free and the immobilized lipase and LPS-2 immobilized PPL had the highest catalytic efficiency in the three. Meanwhile, the LPS supports exhibited many advantages than small porous materials for immobilizing PPL. PMID- 16697483 TI - Fate of hairpin transcript components during RNA silencing and its suppression in transgenic virus-resistant tobacco. AB - Transgenic tobacco plants, carrying a Potato virus Y (PVY)-NIa hairpin sequence separated by a unique unrelated spacer sequence were specifically silenced and highly resistant to PVY infection. In such plants neither PVY-NIa nor spacer transgene transcripts were detectable by specific quantitative real time reverse transcriptase PCR (RT-qPCR) assays of similar relative efficiencies developed for direct comparative analysis. However, small interfering RNAs (siRNAs) specific for the PVY sequence of the transgene and none specific for the LNYV spacer sequence were detected. Following infection with Cucumber mosaic virus (CMV), which suppresses dsRNA-induced RNA silencing, transcript levels of PVY-NIa as well as spacer sequence increased manifold with the same time course. The cellular abundance of the single-stranded (ss) spacer sequence was consistently higher than that of PVY dsRNA in all cases. The results show that during RNA silencing and its suppression of a hairpin transcript in transgenic tobacco, the ssRNA spacer sequence is affected differently than the dsRNA. In PVY-silenced plants, the spacer is efficiently degraded by a mechanism not involving the accumulation of siRNAs, while following suppression of RNA silencing by CMV, the spacer appears protected from degradation. PMID- 16697484 TI - Characterization of Spodoptera exigua multicapsid nucleopolyhedrovirus ORF100 and ORF101, two homologues of E. coli ChaB. AB - ChaB, a putative regulator of ChaA in Escherichia coli (E. coli), and its homologues constitute a multigene family known to occur among bacteria, archaeabacteria and baculoviruses. The baculovirus Spodoptera exigua multicapsid nucleopolyhedrovirus contains two homologues of E. coli ChaB, open reading frames (ORFs) 100 (Se100) and 101 (Se101). RT-PCR and 5' RACE analyses indicated that transcription of both SeMNPV chaB genes occurs by 24 h postinfection and is initiated at a late consensus ATAAG motif. Immunoblot analysis showed that Se100 and Se101 were, respectively, expressed as 11 and 28kDa proteins in infected cells, and they were distributed in both cytoplasm and nucleus. Further analysis revealed that Se100 and Se101 are associated with nuclear structure and only present in the nucleocapsid of occlusion-derived virus. Additionally, column chromatography analysis showed that both proteins could weakly interact with nucleic acids. It was proposed that SeMNPV ChaB might function as DNA binding proteins. PMID- 16697485 TI - Oral bioavailability of a low molecular weight heparin using a polymeric delivery system. AB - Low molecular weight heparins (LMWHs) are the standards of anticoagulant for the prevention of deep vein thrombosis (DVT) in patients undergoing arthroplasty and abdominal surgery. However, LMWHs are so far only administered by parenteral route. Thus, they are usually replaced by oral warfarin for outpatient therapy. Since warfarin has a slow onset and high incidence of drug-drug interaction, there is a great need for the development of an oral LMWH formulation. LMWH (tinzaparin)-loaded nanoparticles prepared with a blend of a polyester and a polycationic polymethacrylate by the double emulsion method were administered orally in fasted rabbits. The plasma tinzaparin concentration was measured by a chromogenic anti-factor Xa assay. After oral administration of two doses of tinzaparin-loaded nanoparticles (200 and 600 anti-Xa U/kg), the oral absorption was observed between 4 and 10 or 12 h, with a delayed onset of action ranging from 3 to 4 h. Mean absolute bioavailabilities were 51% and 59% for the two tested doses. We now report that the encapsulation of tinzaparin into nanoparticles is likely to contribute to its oral efficacy with an anticoagulant effect prolonged up to 8 h. PMID- 16697486 TI - Survey of handheld computing among medical students. AB - The purpose of this study was to identify trends in the utilization and acceptance of handheld computers (personal digital assistants) among medical students during preclinical and clinical training. We surveyed 366 medical students and collected information on computer expertise, current handheld computer use, predicted future use, and user acceptance. Handheld computers were primarily used for personal applications by students during their preclinical training and as drug references and clinical calculators during their clinical training. In the future, all participants predicted they would use handheld computers at significantly higher rates and on a broader range of medical applications. The adoption of handheld computing was independent of user satisfaction. Those with more clinical experience were less satisfied with handheld computers, suggesting that the expectations of the more experienced users were not met. The lack of institutional support was seen as a key limitation. PMID- 16697487 TI - Quality control for handling of accidental blood exposures. AB - A regional counselling service was established to handle all accidental blood exposures using a standardized protocol. Levels of risk were assessed using an algorithm. Accidents that posed a risk for the transmission of hepatitis B (HBV), hepatitis C (HCV) and human immunodeficiency virus (HIV) were classified as 'high risk', whereas accidents that posed a risk for HBV alone were classified as 'low risk'. Medical interventions were implemented according to the level of risk. During a one-year period, all accidents were registered and analysed for adherence to the standard protocol. In 2003, the centre handled 454 incidents. Of these, 36 (7.9%) incidents were assessed as no risk, 329 (72.5%) were assessed as low risk, and 67 (14.8%) were assessed as high risk. Due to incomplete registration, 22 (4.8%) incidents could not be analysed further. In total, 36% of the incidents with risk for HBV transmission and 40% of the incidents with risk for HCV and HIV transmission were not handled according to the proposed protocol. Breaches consisted of over-reaction (25/396) as well as insufficient response (123/396). Potentially inadequate treatment occurred for HIV postexposure prophylaxis in 12 of 63 incidents. Incomplete follow-up for HCV occurred in 11 of 63 incidents, and lack of HBV immunoglobulin administration occurred in five of 396 incidents, including three high-risk incidents. In 21 of 396 low-risk exposures, the breaches in protocol resulted from late reporting. It remains difficult to achieve an acceptable level of standardized care when using standard operational procedures. Documentation and evaluation of flaws are essential to improve the system. PMID- 16697488 TI - NSAIDs and Alzheimer disease: epidemiological, animal model and clinical studies. AB - This review reports correlations between four independent fields related to inflammation and Alzheimer disease: fundamental pathology, epidemiology, transgenic animal studies and clinical trials. Activated microglia, along with a spectrum of inflammatory mediators, have been identified in association with the lesions of Alzheimer disease (AD), suggesting that antiinflammatory agents such as NSAIDs should protect against the disease. In multiple epidemiological investigations testing this hypothesis, a significant risk reduction, or a trend towards such a reduction has been observed in long term as opposed to short term users of traditional NSAIDs. In studies where such NSAIDs have been administered to AD transgenic mice, a dose dependent reduction in pathology was observed. The selective C0X-2 inhibitors were ineffective. Results of clinical investigations have so far been disappointing but have nevertheless correlated with fundamental pathological findings and with transgenic mouse results. Four clinical trials using selective COX-2 inhibitors failed which is in keeping with the animal results and is consistent with pathological findings demonstrating that COX-1 and not COX-2 is the appropriate target in activated human microglia. A low dose trial of the traditional NSAID naproxen also failed, but pilot trials using therapeutically established doses of indomethacin and diclofenac/misoprostol showed promise. Further clinical investigations with relatively high doses of traditional NSAIDs might be warranted, although significant side effects should be anticipated. PMID- 16697489 TI - QSAR models based on quantum topological molecular similarity. AB - A new method called quantum topological molecular similarity (QTMS) was fairly recently proposed [J. Chem. Inf. Comp. Sc., 41, 2001, 764] to construct a variety of medicinal, ecological and physical organic QSAR/QSPRs. QTMS method uses quantum chemical topology (QCT) to define electronic descriptors drawn from modern ab initio wave functions of geometry-optimised molecules. It was shown that the current abundance of computing power can be utilised to inject realistic descriptors into QSAR/QSPRs. In this article we study seven datasets of medicinal interest : the dissociation constants (pK(a)) for a set of substituted imidazolines , the pK(a) of imidazoles , the ability of a set of indole derivatives to displace [(3)H] flunitrazepam from binding to bovine cortical membranes , the influenza inhibition constants for a set of benzimidazoles , the interaction constants for a set of amides and the enzyme liver alcohol dehydrogenase , the natriuretic activity of sulphonamide carbonic anhydrase inhibitors and the toxicity of a series of benzyl alcohols. A partial least square analysis in conjunction with a genetic algorithm delivered excellent models. They are also able to highlight the active site, of the ligand or the molecule whose structure determines the activity. The advantages and limitations of QTMS are discussed. PMID- 16697490 TI - Chemotherapy of leishmaniasis part III: synthesis and bioevaluation of novel aryl substituted terpenyl pyrimidines as antileishmanial agents. AB - Some aryl substituted terpenyl pyrimidines 4 (a-p) have been synthesized using novel synthetic methods. The compounds were screened for in vitro antileishmanial activity against promastigotes. Compounds 4c, 4i and 4l showed IC(50) values as 35, 35 and 25 microg ml(-1). PMID- 16697491 TI - New zirconium (IV) complexes of coumarins with cytotoxic activity. AB - Complexes of zirconium (IV) with some bis-coumarin ligands have been synthesized. The zirconium (IV) complexes with bis-coumarins were characterized by different physicochemical methods-elemental analysis, IR-, and (1)H-NMR-spectroscopies and mass spectral data. The spectral data of zirconium (IV) complexes were interpreted on the basis of comparison with the spectra of the free ligands. The results of the ligands and their complexes, based on spectral data are informative and useful for suggestion of the metal-ligand binding mode. Cytotoxic screening by MTT assay was carried out. In the present study we performed comparative evaluation of the cytotoxic effects of the three newly synthesized zirconium complexes against the acute myeloid leukemia derived HL-60 and the chronic myeloid leukemia LAMA-84. The preliminary cytotoxicity screening program revealed that the investigated zirconium complexes induced 50% inhibition of the cell viability of HL-60 and LAMA-84 cells at micromolar concentrations and thus could be considered as biologically active. Independently of the tumor test system evaluated the complex of bis(4-hydroxy-2-oxo-2H-chromen-3-yl)-(1H-pyrazol 3-yl)-methane proved superior to the remaining agents with respect to the IC(50) values obtained. The complexes of both the other coumarins evaluated proved to be less potent than the corresponding free ligands, as evidenced by the IC(50) values obtained. Thus the zirconium complexes with coumarin ligands represent a novel class of antiproliferative agents, which deserve further attention in search of anticancer lead compounds. PMID- 16697492 TI - [A painful foot]. PMID- 16697493 TI - [Allergy emergencies, management and prevention]. PMID- 16697494 TI - Effect of docosahexaenoic acid-rich fish oil supplementation on human leukocyte function. AB - BACKGROUND: The effect of a docosahexaenoic acid (DHA)-rich fish oil (FO) supplementation on human leukocyte function was investigated. METHODS: Ten male volunteers were supplemented with 3g/day FO containing 26% eicosapentaenoic acid (EPA, 20:5, n-3) and 54% DHA (22:6, n-3) for 2 months. RESULTS: FO supplementation changed the fatty acid (FA) composition of leukocytes resulting in an increase of n-3/n-6 ratio from 0.18 to 0.62 in lymphocytes and from 0.15 to 0.70 in neutrophils. DHA-rich FO stimulated an increase in phagocytic activity by 62% and 145% in neutrophils and monocytes, respectively. Neutrophil chemotactic response was increased by 128%. The rate of production of reactive oxygen species by neutrophils was also increased, as it was with lymphocyte proliferation. These changes were partially reversed after a 2-month wash out period. With respect to cytokine production by lymphocytes, interleukin (IL)-4 release was not altered, whereas secretions of IL-10, interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha were raised. These results are in contrast to those described by others using EPA-rich FO supplementation. Lymphocyte pleiotropic gene expression was analyzed by a macroarray technique. Of the analyzed genes (588 in total), 77 were modified by the supplementation. FO supplementation resulted in up regulation of 6 genes (GATA binding protein 2, IL-6 signal transducer, transforming growth factor alpha, TNF, heat shock 90kDa protein 1-alpha and heat shock protein 70kDa 1A) and a down regulation of 71 genes (92.2% of total genes changed). The largest functional group of altered genes was that related to signaling pathways (22% of the total modified genes). CONCLUSIONS: Therefore, although EPA and DHA are members of n-3 FA family, changes in the proportion of DHA and EPA exert different effects on neutrophil, monocyte and lymphocyte function, which may be a result of specific changes in gene expression. PMID- 16697495 TI - ESPEN Guidelines on Enteral Nutrition: Adult renal failure. AB - Enteral nutrition (EN) by means of oral nutritional supplements (ONS) and tube feeding (TF) offers the possibility of increasing or ensuring nutrient intake in cases where normal food intake is inadequate. These guidelines are intended to give evidence-based recommendations for the use of ONS and TF in nephrology patients. They were developed by an interdisciplinary expert group in accordance with officially accepted standards and are based on all relevant publications since 1985. They were discussed and accepted in a consensus conference. Because of the nutritional impact of renal diseases, EN is widely used in nephrology practice. Patients with acute renal failure (ARF) and critical illness are characterized by a highly catabolic state and need depurative techniques inducing massive nutrient loss. EN by TF is the preferred route for nutritional support in these patients. EN by means of ONS is the preferred way of refeeding for depleted conservatively treated chronic renal failure patients and dialysis patients. Undernutrition is an independent factor of survival in dialysis patients. ONS was shown to improve nutritional status in this setting. An increase in survival has been recently reported when nutritional status was improved by ONS. PMID- 16697496 TI - Metabolic effects of parenteral nutrition enriched with n-3 polyunsaturated fatty acids in critically ill patients. AB - BACKGROUND & AIMS: n-3 fatty acids are expected to downregulate the inflammatory responses, and hence may decrease insulin resistance. On the other hand, n-3 fatty acid supplementation has been reported to increase glycemia in type 2 diabetes. We therefore assessed the effect of n-3 fatty acids delivered with parenteral nutrition on glucose metabolism in surgical intensive care patients. METHODS: Twenty-four surgical intensive care patients were randomized to receive parenteral nutrition providing 1.25 times their fasting energy expenditure, with 0.25 g of either an n-3 fatty acid enriched-or a soy bean-lipid emulsion. Energy metabolism, glucose production, gluconeogenesis and hepatic de novo lipogenesis were evaluated after 4 days. RESULTS: Total energy expenditure was significantly lower in patients receiving n-3 fatty acids (0.015+/-0.001 vs. 0.019+/-0.001 kcal/kg/min with soy bean lipids (P<0.05)). Glucose oxidation, lipid oxidation, glucose production, gluconeogenesis, hepatic de novo lipogenesis, plasma glucose, insulin and glucagon concentrations did not differ (all P>0.05) in the 2 groups. CONCLUSIONS: n-3 fatty acids were well tolerated in this group of severely ill patients. They decreased total energy expenditure without adverse metabolic effects. PMID- 16697497 TI - Rice protein-induced enterocolitis syndrome. AB - BACKGROUND AND AIMS: Food protein-induced enterocolitis syndrome (FPIES) is a non IgE-mediated allergic reaction, usually to cow's milk or soy protein. The aim is to present a series of infants with enterocolitis syndrome developed after rice ingestion. To our knowledge, the issue has only once been described in patients from Europe. METHODS: Data on five patients were retrospectively analyzed. The patients presented with vomiting, diarrhea and dehydration; therefore, allergy was not considered as a diagnosis and the patients underwent broad diagnostic evaluation. Finally, an open food challenge was performed for suspicion of rice allergy. RESULTS: Detailed clinical and laboratory findings are presented. As all patients had symptoms which could have been indicative of a broad spectrum of diseases, a median of two hospitalizations was needed to establish the diagnosis. When the rice protein-induced enterocolitis syndrome is suspected all patients should undergo open food challenge test, as no other diagnostic procedure can confirm the diagnosis. CONCLUSION: This report shows that even hypoallergenic foods such as rice may cause FPIES and should be considered in the differential diagnosis of profuse vomiting and prostration in infants introduced to some kind of rice protein. PMID- 16697499 TI - Post-oesophagectomy early enteral nutrition via a needle catheter jejunostomy: 8 year experience at a specialist unit. AB - BACKGROUND AND AIMS: The purpose of this study was to prospectively evaluate post operative jejunostomy feeding in terms of nutritional, biochemical, gastrointestinal and mechanical complications in patients undergoing upper gastrointestinal surgery for oesophageal malignancy. METHODS: The study included 205 consecutive patients who underwent oesophagectomy for malignancy. All patients had a needle catheter jejunostomy (NCJ) inserted at the conclusion of laparotomy. Patients were followed prospectively to record nutritional intake, type of feed administered, rate progression, tolerance, weight changes and complications either mechanical, biochemical or gastrointestinal. RESULTS: Ninety two per cent of patients were successfully fed exclusively by NCJ post oesophagectomy, and 94% of patients were tolerating a maintenance regimen of 2000 ml feed over 20 h by day 2 post-operatively. Patients spent a median of 15 days on jejunostomy feeding post-surgery (range 2-112 days); however, 26% required prolonged jejunostomy feeding (>20 days). Minor gastrointestinal complications were effectively managed by slowing the rate of infusion, or administering medication. Three (1.4%) serious complications of jejunostomy feeding occurred, all requiring re-laparotomy, one resulting in death. NCJ feeding was extremely effective in preventing severe post-operative weight loss in the majority of oesophagectomy patients post-op. However, oral intake was generally poor at discharge with only 65% of requirements being met orally. Sixteen patients (8%) patients required home jejunostomy feeding. By the first post-operative month, a further 6% (12) patients were recommenced on jejunostomy feeding. CONCLUSION: NCJ feeding is an effective method of providing nutritional support post oesophagectomy, and allows home support for the subset that fail to thrive. Serious complications, most usually intestinal ischaemia or intractable diarrhoea, are rare. PMID- 16697500 TI - ESPEN Guidelines on Enteral Nutrition: Non-surgical oncology. AB - Enteral nutrition (EN) by means of oral nutritional supplements (ONS) and tube feeding (TF) offers the possibility of increasing or ensuring nutrient intake in cases where normal food intake is inadequate. These guidelines are intended to give evidence-based recommendations for the use of ONS and TF in cancer patients. They were developed by an interdisciplinary expert group in accordance with officially accepted standards, are based on all relevant publications since 1985 and were discussed and accepted in a consensus conference. Undernutrition and cachexia occur frequently in cancer patients and are indicators of poor prognosis. EN should be started if undernutrition already exists or if food intake is markedly reduced for more than 7-10 days. Standard formulae are recommended for EN. Nutritional needs generally are comparable to non-cancer subjects. In cachectic patients metabolic modulators such as progestins, steroids and possibly eicosapentaenoic acid may help to improve nutritional status. EN is indicated preoperatively for 5-7 days in cancer patients undergoing major abdominal surgery. During radiotherapy of head/neck and gastrointestinal regions dietary counselling and ONS prevent weight loss and interruption of radiotherapy. Routine EN is not indicated during (high-dose) chemotherapy. PMID- 16697501 TI - A completely nurse-driven PICC-line service in a surgical ward: a prospective audit of the implementation-phase in unselected patients. PMID- 16697502 TI - Managing the patient journey through enteral nutritional care. AB - Nutritional support provision does not happen by accident. Clinical dimensions include screening and assessment, estimation of requirements, identification of a feeding route and the subsequent need for monitoring. Patients may need different forms of nutritional intervention during the course of their illness. Furthermore, these may need to be provided in different locations as their clinical status changes. If this is not properly managed there is potential for inappropriate treatment to be given. Clinical processes can only be effectively implemented if there is a robust infrastructure. The clinical team need to understand the different elements involved in effective service provision and this depends on bringing together disciplines which do not feature overtly on the clinical agenda including catering, finance and senior management. Excellent communication skills at all levels, financial awareness and insight into how other departments function are fundamental to success. Practice needs to be reviewed constantly and creativity about all aspects of service delivery is essential. Finally, it is important that key stakeholders are identified and involved so that they can support any successes and developments. This will raise awareness of the benefits of nutritional intervention and help to ensure that the right resources are available when they are needed. PMID- 16697503 TI - Maternal intranasal immunization with outer membrane protein P6 maintains specific antibody level of derived offspring. AB - Acute otitis media is one of the most common infectious diseases in children younger than 2 years of age. Immunological studies in young children have revealed that immature antibody-responses to major pathogens, such as S. pneumoniae, H.influenzae, would cause the vulnerability to upper respiratory tract infections. Thus, it is very important to induce effective protective immunity among children younger than 2 years of age. In this study, we evaluated the capacity of maternal immunization with P6 of H. influenzae to evoke specific antibody to P6 and to transfer it to offspring. We intranasally immunized mother mice with P6 and investigated the induction of specific antibody in sera and breast milk. The specific antibody among offspring delivered by immunized mother was also investigated according to the nursing status to evaluate the importance of breast feedings by immunized mothers. Anti-P6 specific IgG in sera were high at delivery and maintained during nursing periods among P6-immunized mother mice. Anti-P6 specific IgG were predominantly induced in breast milk. IgG subclass induced in sera and breast milk from P6-immunized mother mice were IgG2b, followed by IgG1 and IgG2a subclass. Offspring delivered by P6-immunized mothers had anti-P6 specific IgG in sera at the birth. The levels of anti-P6 specific IgG in sera from offspring breast-fed by P6-immunized mothers were then increased until day 14 and then decreased on day 21. The anti-P6 specific IgG in sera from offspring breast-fed by sham-immunized mothers were rapidly decreased after birth. The current findings strongly suggest that maternal intranasal immunization with P6 would be an attractive strategy against NTHi infections during early childhood. It can supply protective antibodies via transplacental transfer during pregnancy and via breast milk after birth. PMID- 16697504 TI - Introduction to DNA vaccines issue. PMID- 16697505 TI - The cellular effect of a single interrupted suture on tendon. AB - The effects on cell and matrix morphology of a single interrupted suture are described in rabbit (vascular) and mouse (avascular) digital flexor tendons. This model of tendon injury is reproducible and suitable for quantitative histological analysis. Tendons analysed at day 1, 3, 5, 7 and 14 after wounding demonstrated a well-demarcated "acellular zone" around the suture within 24 hours and persisting over 14 days. The placement of an untied suture in tendon did not produce this effect but tying and releasing the tied knot did. The rapidity of onset suggests that cells move from the zone of injury into less mechanically strained tissue. The acellular zone was apparent in rabbit hind paw flexor tendon which is vascularised and the corresponding tendon in mouse which has no intrinsic blood vessels. This phenomenon highlights biological events that must be considered in parallel with the current trend for multistrand locking flexor tendon suture repairs. PMID- 16697506 TI - Re: arthroscopic triangular fibrocartilage complex debridement using radiofrequency probes, and arthroscopic electrothermal collagen shrinkage for symptomatic laxity of the scapholunate interosseous ligament. PMID- 16697507 TI - Further evidence of the effects of global warming on lichens, particularly those with Trentepohlia phycobionts. AB - Increasing evidence suggests that lichens are responding to climate change in Western Europe. More epiphytic species appear to be increasing, rather than declining, as a result of global warming. Many terricolous species, in contrast, are declining. Changes to epiphytic floras are markedly more rapid in formerly heavily polluted, generally built-up or open rural areas, as compared to forested regions. Both the distribution (southern) and ecology (warmth-loving) of the newly established or increasing species seem to be determined by global warming. Epiphytic temperate to boreo-montane species appear to be relatively unaffected. Vacant niches caused by other environmental changes are showing the most pronounced effects of global warming. Species most rapidly increasing in forests, although taxonomically unrelated, all contain Trentepohlia as phycobiont in addition to having a southern distribution. This suggests that in this habitat, Trentepohlia algae, rather than the different lichen symbioses, are affected by global warming. PMID- 16697508 TI - Nutritional status and spousal empowerment among native Amazonians. AB - Researchers and development organizations have shown interest in individual empowerment because it presumably improves well-being. Estimates of empowerment's effects on well-being contain biases from the potential endogeneity of empowerment. Using data from a sexually egalitarian and highly autarkic society of foragers and horticulturalists in the Bolivian Amazon, the Tsimane', we overcome the problems that this poses by: (1) matching spouses' responses to the same questions about who makes decisions or who breaks ties in 10 domains to improve accuracy in measures of empowerment; and (2) using parental attributes of spouses as instrumental variables for spousal empowerment. Outcomes include two anthropometric indices of short-run nutritional status: body-mass index and age and sex-standardized z scores of mid-arm muscle area. The amount of empowerment of household heads did not affect their nutritional status or other indicators of their well-being, such as income, wealth, expenditures, happiness, social capital, or self-perceived health. It also did not affect the nutritional status of their offspring. Nor did it affect the difference in income, wealth, or monetary expenditures between spouses. The insubstantial effects persisted with other definitions of empowerment or types of regressions. We end with a discussion of why empowerment, despite its popularity in development discourse, has such tenuous links with objective indicators of well-being, and the implication of this finding for future studies of empowerment's effects. PMID- 16697509 TI - The close relation between birth, abortion and employment rates in Sweden from 1980 to 2004. AB - Birth and abortion rates in Sweden have fluctuated since 1980 while the proportion between the rates are the same at the beginning and end of the period. An increase in birth rates in the late 1980s resulted in a peak in 1991 and 1992, with 124,000 live births each year. Thereafter followed a steady decline in the rate until 2000, when the number of live births was about 90,000. At that point, the trend changed to an increase. The aim of this analysis was to investigate any relation between employment rates and the number of live births among women aged 20-34, and at the same time to explore the trend for abortion rates compared to the trend for live births. The relation between employment status and live birth rate is statistically more significant for women than men, and the rates have a higher correlation for the period after 1986. Young adults in this age group are vulnerable to economic cycles that can explain this covariation but the decline in birth rates in economically developed societies has multidimensional aspects and many other possible explanations. Much has been done in recent years in Sweden to decrease household inequality for families with children to avoid the risk of relative poverty, but the fact that there is no explicit health policy to reduce the abortion level that remain unchanged since the early 1980s may appear as a notable lack of strategy in a country with many other health-related goals. PMID- 16697510 TI - Making the normal deviant: the introduction of predictive medicine in life insurance. AB - Over the past years, one of the most discussed topics in policy debates on genetics has been the use of genetic testing in insurance. Many of these debates have been rather speculative and abstract. In a recent contribution to this journal, Kaufert therefore urged for "a proper research agenda" to study the issue, arguing for the need of anthropological and sociological research of the insurance world. This article will make a start with this. Based on ethnographic fieldwork in two Belgium insurance companies, this study analyses the ways insurers account for predictive medicine (lifestyle, genetics) during underwriting. We demonstrate how insurers highlight predictive lifestyle health information and how this articulates with a fault based approach in underwriting. Individual responsibility for health risks becomes the golden standard for assessing one's fitness for membership of the insurance pool. Moreover, these developments imply a changed concept of "normal standard" in insurance, increasing the conditions to fulfil to be part of the insurance group. Predictive medicine constitutes new ground in the old debates about individual control, responsibility and blame for health. This goes to the heart of the basis for citizenship and how this articulates with membership--or, if you want, exclusion- of the insurance pool. PMID- 16697511 TI - Is chronic widespread pain biographically disruptive? AB - This article draws on findings from a study of eight people (aged 40-60) with chronic widespread pain and their families, living in the West Midlands area of the UK. Data were generated through a series of in-depth interviews, based on a lifegrid and on participants' diaries. We explore the experience of chronic widespread pain in the context of sufferers' biographies, examining how people attempt to account for and give meaning to their pain onset and development and how they remake identity following disruption to their lives. We use these accounts to consider whether chronic widespread pain is biographically disruptive, or whether any of the alternative notions better describe the experience. We conclude that the consideration of context in biographical disruption should extend to lifestage and perceived life expectancy. PMID- 16697512 TI - Should structural interventions be evaluated using RCTs? The case of HIV prevention. AB - Structural interventions addressing macro-social determinants of risk have been suggested as potentially important adjuncts to biomedical and behavioural interventions for the prevention of HIV and other diseases. A few interventions of this type have been evaluated using randomised controlled trials (RCTs), the most rigorous design to evaluate the effects of biomedical and behavioural interventions. The appropriateness of applying RCTs to structural interventions is however debated. This paper considers whether issues of ethics, feasibility and utility preclude the use of RCTs in evaluations of structural interventions for HIV prevention. We conclude there is nothing particular to this category of interventions prohibiting use of RCTs. However, we suggest that RCTs may prove unacceptable, unfeasible or not useful in certain circumstances, such as where an intervention brings important benefits other than HIV prevention (such as increased income); where leaders of clusters do not allow decisions about macro social policies to be determined randomly; where the unit of social organization addressed by an intervention is so large that recruitment of adequate numbers of clusters is impossible; and where the period required to trial interventions extends beyond practical decision-making time-scales. In such cases, alternative evaluative designs must be assessed for their ability to provide evidence of intervention effectiveness. PMID- 16697513 TI - Age-related changes in duration reproduction: involvement of working memory processes. AB - The aim of the present research was to study age-related changes in duration reproduction by differentiating the working memory processes underlying this time estimation task. We compared performances of young and elderly adults in a duration reproduction task performed in simple and concurrent task conditions. Participants were also administered working memory tests to measure storage and central executive functions. Findings indicated a differential involvement of working memory storage and central executive functions in age-related differences in temporal tasks. The limited storage capacities explained age-related changes in the simple task of duration reproduction, and the dysfunctioning of central executive functions accounted for age-related changes in duration reproduction performed in a concurrent task condition, which involves greater attentional resources. PMID- 16697514 TI - Differential polyamine analogue effects in four human breast cancer cell lines. AB - Polyamine analogues have demonstrated anti-tumour activity in a number of solid tumour models. In the present study we compared the cytotoxicities of three polyamine analogues against four breast cancer cell lines. All cell lines are derived from tumours of women with breast cancer and, although we are sampling just a small number of tumours, they represent a spectrum of the genetic plethora of breast cancers. Cytotoxicity, over a dose range from 0.1 to 100 microM, was evaluated with three different cytotoxicity assays performed in 96-well plates. Comparing the effects of the analogues on polyamine pools with data from the cytotoxicity assays indicates that there was not a direct correlation between polyamine pool depletion and cytotoxicity. Flow cytometry was used to investigate analogue-induced cell death as measured by the appearance of a sub-G(1) peak. Induction of cell death by the analogues differed in the cell lines, however, cell death when induced was apoptotic, as demonstrated by detection of apoptotic bodies with immunofluorescence microscopy of propidium iodide-stained nuclei. Comparing the flow cytometry-derived data and the data from the cytotoxicity assays reveals that the analogues exert their effects by inhibiting cell growth and/or inducing cell death. PMID- 16697515 TI - Direct observation of protein folding in nanoenvironments using a molecular ruler. AB - We observe folding of horse heart cytochrome c in various environments including nano-compartments (micelles and reverse micelles). Using picosecond-resolved Forster resonance energy transfer (FRET) dynamics of an extrinsic covalently attached probe dansyl (donor) at the surface of the protein to a heme group (acceptor) embedded inside the protein, we measured angstrom-resolved donor acceptor distances in the environments. The overall structural perturbations of the protein revealed from the FRET experiments are in close agreement with our circular dichroism (CD) and dynamic light scattering (DLS) studies on the protein in a variety of solution conditions. The change of segmental motion of the protein due to imposed restriction in the nano-compartments compared to that in bulk buffer is also revealed by temporal fluorescence anisotropy of the dansyl probe. PMID- 16697516 TI - Properties of microsolvated ions: from the microenvironment of chromophore and alkali metal ions in proteins to negative ions in water clusters. AB - Here we discuss the fascinating chemistry and physics of microsolvated ions that bridge the transition from bare ions in gas phase to ions in solution. Such ions occur in many situations in biochemistry and are crucial for several functions; metal ions, for example, must remove their water shell to pass through ion pumps in membranes. Furthermore, only a few water molecules are buried in the hydrophobic pockets of proteins where they are bound to charged amino acid residues or ionic chromophores. Another aspect is the reactivity of microsolvated ions and the importance in atmospheric, organic and inorganic chemistry. We close by a discussion of the stability of molecular dianions, and how hydration affects the electronic binding energy. There is a vast literature on microsolvated ions, and in this review we are far from being comprehensive, rather we mainly bring examples of our own work. PMID- 16697517 TI - Water quality modelling of the river Yamuna (India) using QUAL2E-UNCAS. AB - This paper describes the utility of QUAL2E as a modelling package in the evaluation of a water quality improvement programme. In this study, QUAL2E was applied to determine the pollution loads in the river Yamuna during its course through the national capital territory of Delhi, India. The study aimed at examining the influence of different scenarios on river water quality. Four different pollution scenarios were analysed besides the 'business as usual' situation. The study revealed that it was necessary to treat the discharge from drains to the river Yamuna and diversion of a substantial load to the Agra canal for further treatment was also essential. It was also established through this study that maintaining a flow rate of more than 10 m(3)/s in the river could also help preserve the river's water quality. To clearly display the model outcomes and demarcate polluted zones in the river stretch, model results were interfaced with a Geographical Information System (GIS) to produce cartographic outputs. In addition, uncertainty analysis in the form of first-order error analysis and Monte Carlo analysis was performed, to realise the effect of each model parameter on DO and BOD predictions. The uncertainty analysis gave satisfactory results on simulated data. PMID- 16697518 TI - Product quality-based eco-efficiency applied to digital cameras. AB - When calculating eco-efficiency, there are considerable confusion and controversy about what the product value is and how it should be quantified. We have proposed here a quantification method for eco-efficiency that derives the ratio of the multiplication value of the product quality and the life span of a product to its whole environmental impact based on Life Cycle Assessment (LCA). In this study, product quality was used as the product value and quantified by the following three steps: (1) normalization based on a value function, (2) determination of the subjective weighting factors of the attributes, and (3) calculation of product quality of the chosen products. The applicability of the proposed method to an actual product was evaluated using digital cameras. The results show that the eco-efficiency values of products equipped with rechargeable batteries were higher than those products that use alkaline batteries, because of higher quality values and lower environmental impacts. The sensitivity analysis shows that the proposed method was superior to the existing methods, because it enables to identify the quality level of the chosen products by considering all products that have the same functions in the market and because, when adding a new product, the calculated quality values in the proposed method do not have to be changed. PMID- 16697519 TI - Effect of intravesical Resiniferatoxin (RTX) on lower urinary tract symptoms, urodynamic parameters, and quality of life of patients with urodynamic increased bladder sensation. AB - OBJECTIVES: Intravesical resiniferatoxin (RTX) has been used with variable efficacy in the treatment of detrusor overactivity (DO). Patients with interstitial cystitis (IC) failed to benefit from this treatment, but a single placebo-controlled study in patients with non-IC painful bladders showed that RTX was effective in the short-term. We investigated the efficacy of intravesical RTX in patients with urgency and frequency due to increased bladder sensation. METHODS: Patients with intractable urgency and frequency, with or without urgency incontinence or bladder pain/discomfort, and with no urodynamic evidence of DO were recruited. After a single intravesical instillation of 100ml 50 nM RTX solution, patients were followed at 1, 3, and 6 mo for changes in urodynamics, bladder diary, the King's Health Questionnaire (KHQ), and degree of bladder pain. RESULTS: Fifteen patients (mean age, 52.5 yr) were treated. RTX significantly improved maximum cystometric capacity, volume at first desire to void, mean micturition volume, 24-h frequency, and daytime frequency for up to 6 mo after treatment. The overall KHQ score improved at all time points, with sustained improvements in the Symptom Severity, Incontinence Impact, and Personal Relationships domains. A >50% decrease in pain was reported by five of seven patients with painful bladders at 1 mo, but only one of seven at 6 mo. CONCLUSIONS: In our small open-label study, a single administration of intravesical RTX in patients with frequency and urgency due to increased bladder sensation significantly improved lower urinary tract symptoms, urodynamic parameters, and quality of life for up to 6 mo. PMID- 16697520 TI - A (-5, -7) proPSA based artificial neural network to detect prostate cancer. AB - OBJECTIVE: The pro-forms of prostate specific antigen (-2,-5,-7 proPSA) and also %free PSA based artificial neural networks (ANN) have been suggested to enhance the discrimination between prostate cancer (PCa) and no evidence of malignancy (NEM). This study reports on the combined use of proPSA within a %free PSA based ANN to enhance specificity of PCa. METHODS: Serum samples from 898 patients with PCa (n=384) or NEM (n=514) within the PSA range 1-10 microg/l were analyzed for PSA, free PSA and (-5,-7) proPSA (Roche assays). Patient data from two centers - taken first from the Swiss site of the ERSPC (Aarau) and from a referral population (Berlin) have been analyzed. Leave-one-out ANN models with the variables PSA, %fPSA, proPSA, prostate volume and status of digital rectal examination (DRE) were constructed and compared by receiver-operating characteristic (ROC) curve analysis. RESULTS: (-5,-7) proPSA was only significantly different between NEM and PCa in the PSA range 4-10 microg/l. Within the PSA range 4-10 microg/l (Berlin group) the ANN including only the two variables %fPSA and proPSA could reach the same performance like the conventional ANN with PSA, %fPSA, age, prostate volume and DRE (both AUCs: 0.84) However, at 95% sensitivity all ANN could not improve specificity compared to %fPSA. CONCLUSIONS: ProPSA as single parameter did not improve specificity over %fPSA whereas proPSA and %fPSA within an ANN in the PSA range 4-10 microg/l substituted prostate volume and DRE. At 95% sensitivity only ANN with prostate volume and DRE perform significantly better than %fPSA. PMID- 16697521 TI - Prognostic role of the histologic subtypes of renal cell carcinoma after slide revision. AB - OBJECTIVES: To evaluate concordance, using the kappa statistic, between tumour histotypes assigned at initial diagnosis and those after slide revision, and to assess the independent predictive value of reviewed tumour histotype. MATERIALS AND METHODS: We analysed records of 491 patients who had undergone partial or radical nephrectomy for renal cell carcinoma (RCC) in our department from 1986 to 2000. Pathologic slides were reviewed by one uropathologist blinded to original diagnoses and follow-up data, who reassigned histologic subtype and nuclear grades. RESULTS: Concordance between original and reviewed histotypes was substantial (kappa = 0.52). Data were affected by the year of initial diagnosis: Concordance was moderate (kappa = 0.43) in patients treated from 1986 to 1997 and substantial (kappa = 0.73) in those observed thereafter. The original histotype did not stratify the cancer-related outcome. According to the reviewed histotypes, the 5-year cancer-specific survival probabilities of conventional, papillary and chromophobe RCC patients were 81.3%, 90.1% and 100%, respectively (p = 0.009). Patients with conventional RCC showed significantly lower cancer specific survival than those with papillary or chromophobe RCC, even in the cases with locally advanced (p = 0.04) or high-grade (p = 0.02) neoplasms. The reviewed histologic subtype was not an independent prediction of cancer-specific survival on multivariate analysis. CONCLUSION: Concordance between histologic subtype assigned before 1997 and after slide revision was only moderate. In univariate analysis, reviewed tumour histotype stratified cancer-related outcome, but the data of multivariate analysis were strongly influenced by the prevalent number of conventional RCC. PMID- 16697522 TI - Role of protein kinase Calpha in melatonin signal transduction. AB - Melatonin induces nuclear exclusion of the androgen receptor (AR) via activation of protein kinase C (PKC). The specific members of the PKC superfamily involved in AR nuclear exclusion were investigated in prostate cancer PC3 cells stably transfected with the wild-type androgen receptor (PC3-AR). PKCalpha was essentially cytoplasmic whereas PKCbeta and PKCepsilon were essentially membranal, suggesting their constitutive activity in the PC3-AR cells. Melatonin treatment induced membrane association of PKCalpha in a time and dose dependent manner. The PKCalpha and PKCbeta1 specific inhibitor GO6976 and the PKCbeta isoform-specific inhibitor hispidin had no effects on AR localization under basal conditions. However, GO6976 but not hispidin negated the melatonin-mediated nuclear exclusion of the AR. These data indicate that PKCalpha activation is a critical step in AR nuclear exclusion by melatonin. They also imply that PKCalpha activation is a potentially effective way to control of the AR activity in prostate cancer cells. PMID- 16697523 TI - Photoelectrocatalytic oxidation of remazol turquoise blue and toxicological assessment of its oxidation products. AB - The ability of photoelectrocatalytic oxidation to degrade the commercially important copper-phtalocyanine dye, remazol turquoise blue 15 (RTB) was investigated. The best experimental condition was optimized, evaluating the performance of Ti/TiO2 thin-film electrodes prepared by sol-gel method in the decolourization of 32 mg L(-1) RTB dye in 0.5 mol L(-1) Na2SO4 pH 8 and applied potential of +1.5 V versus SCE under UV irradiation. Spectrophotometric measurements, high performance liquid chromatography, dissolved organic carbon (TOC) evaluation and stripping analysis of yielding solution obtained after 3 h of photoelectrolysis leads to 100% of absorbance removal from wavelength of 250 800 nm, 79.6% of TOC reduction and the releasing of up to 54.6% dye-bound copper (0.85 mg L(-1)) into the solution. Both, original and oxidized dye solution did not presented mutagenic activity with the strains TA98 and TA100 of Salmonella in the presence and absence of S9 mix at the tested doses. Nevertheless, the yielding photoelectrocatalytic oxidized solution showed an increase in the acute toxicity for Vibrio fischeri bacteria, explained by copper liberation during treatment. PMID- 16697524 TI - Anatomical changes in selected cardio-respiratory brainstem nuclei following early post-natal chronic intermittent hypoxia. AB - Early post-natal environmental exposures, including chronic intermittent hypoxia (CIH), may lead to long-term alterations in cardio-respiratory control, such as reductions in baroreflex sensitivity and acute hypoxic ventilatory responses in adult rats. Although the mechanisms underlying CIH-induced functional metaplasticity are unclear, anatomical alterations within selected brainstem nuclei may develop after CIH. To examine this issue, male rats were exposed to CIH (RAIH) or room air (RARA) for the first 30 days of life and were microinjected unilaterally in the right nodose ganglion with the neuronal tracer tetramethylrhodamine-dextran (TMR-D) to label brainstem neurons receiving vagal and glossopharyngeal projections. Substantial reductions in labeled afferents within the nucleus tractus solitarii (nTS) and significant increases in the total number of labeled neurons within the ventrolateral medulla (VLM), principally in the nucleus ambiguus (Namb; p<0.01) occurred in RAIH. Furthermore, 5-bromo 2'deoxyuridine labeling revealed enhanced neurogenesis within the Namb in RAIH and could partially account for the increased neuronal population in Namb. Thus, CIH-associated cardio-respiratory metaplasticity is accompanied by substantial structural changes within both the nTS and Namb. PMID- 16697526 TI - Neural correlates of serial abacus mental calculation in children: a functional MRI study. AB - Abacus experts have demonstrated extraordinary potential of mental calculation by using an imaginary abacus. But the neural correlates of abacus mental calculation and the imaginary abacus still remain unclear. Here, we report, respectively, the analysis of fMRI images of abacus experts and non-experts in response to the performance of simple and complex serial calculation by visual stimuli as well as the images of the abacus experts with performance of the same tasks by auditory stimuli. We found that activated areas were quite different between two groups. In experts, enhanced activations were mainly observed in fronto-temporal circuit (lateral premotor cortex (LPMC) and posterior temporal areas) in simple addition, but in fronto-parietal circuit (lateral premotor cortex (LPMC) and posterior superior parietal lobe (PSPL)) in complex one. By contrast, in controls, the activated areas were almost similar in both simple and complex tasks, including bilateral inferior parietal lobule, prefrontal and premotor cortices. Furthermore, visual and auditory stimuli generated almost similar activations in experts. These observations reveal that (1) abacus mental calculation induces special patterns of brain response, and simple and complex tasks are sustained by dissociated brain circuits between the temporal and parietal cortices, respectively; (2) the abacus mental calculation may rely on neural resources of visuospatial representations with a super-modal form of abacus beads; (3) the posterior temporal areas and PSPL may be recruited for imaginary abacus. PMID- 16697525 TI - The physiologically modulated electrode potentials at the depth electrode-brain interface in humans. AB - To study the modulated electrical potential specifically related to the electrode brain interface (EBI) in deep brain stimulation (DBS) under physiological condition, we quantitatively identified the physiologically modulated electrode potentials by decomposing the local field potentials (LFPs) recorded from 11 patients (18 electrodes in four different brain regions) who underwent DBS, and correlated them with simultaneously recorded physiological signals of blood pressure (BP) and respiration. Results showed that electrode potentials were modulated by BP and respiration and could be detected as a specific component of the compound LFP signals with a mean (+/-S.D.) amplitude of 6.9+/-1.7 microV. The detection rate and amplitude of the modulated electrode potentials were independent from brain regions and neurological disorders. The current approach can be used to study the changes in properties of the EBI under physiological condition and to investigate the effects of the EBI on the 'crossing' current of either the neural signals to be recorded or the electrical pulses for neurostimulation. PMID- 16697527 TI - Yohimbine acts as a putative in vivo alpha2A/D-antagonist in the rat prefrontal cortex. AB - Yohimbine has been widely used as alpha2-adrenergic receptor antagonist in neurophysiological research and in clinical therapy. In this study, we provide in vivo electrophysiological evidence, that microiontophoretic application of yohimbine (YOH) inhibits spontaneous activity of prefrontal neurons of the rat. By microiontophoretic application of the alpha2A-receptor antagonist BRL44408 (BRL), the effects of YOH could be mimicked, indicating that the action of YOH is manifested through alpha2A/D-receptor mechanisms. Furthermore, the inhibiting effects of YOH or BRL were blocked by alpha2B-receptor antagonist imiloxan. In concert with previous microiontophoretic data, the present findings suggest that alpha2-receptor antagonist YOH predominantly acts on the alpha2A/D-receptor subtype in vivo. Furthermore, we hypothesize that this action is manifested via deactivation of autoreceptors causing increased norepinephrine release, finally inhibiting postsynaptic neurons through the activation of alpha2B-receptors. PMID- 16697528 TI - Disrupted central somatosensory processing in CRPS: a unique characteristic of the syndrome? PMID- 16697529 TI - Influence of genotype, dose and sex on pruritogen-induced scratching behavior in the mouse. AB - Itch features considerable interindividual variability in humans, and initial studies using animal models have demonstrated a likely role of genetic factors in mediating such variability. In an attempt to systematically study genetic mediation of itch in the mouse such that gene identification by linkage mapping might be achieved, we examined scratching behavior induced by histamine and chloroquine in mice of 11 inbred mouse strains. Multiple chloroquine drug doses were used, revealing the existence of inverted-U dose-response relationships in every strain, allowing us to determine strain-dependent peak scratching behavior over the entire dose range. Peak chloroquine-induced scratching varied by 2.5 fold in this set of strains; scratching behavior shows moderate heritability in the mouse. The present data also reveal, for the first time, significant sex differences in pruritogen-induced scratching behavior, with female mice scratching an average of 23% more than males. Finally, a comparison of the strain means obtained here with previously collected data using nociceptive assays revealed a suggestive negative genetic correlation between chloroquine-induced itch and thermal pain, such that strains sensitive to pain are resistant to itch and vice versa. This finding may have implications both for our understanding of itch pathophysiology and for the identification of itch-related genes. PMID- 16697531 TI - Neospora caninum antibodies in commercial fetal bovine serum. AB - The protozoan parasite Neospora caninum is a major cause of abortion in cattle throughout the world. In the process of propagating Neospora in vitro and producing specific antibodies for development of diagnostic assays in the food supply, our laboratory identified the presence of bovine antibodies to N. caninum in fetal bovine sera. The sera were produced commercially and preferentially recommended for tissue culture use and monoclonal antibody production. Seventeen different fetal bovine serum samples of different grades and from four different companies were examined for the presence of total IgG, IgG1, IgG2, and IgM specific for N. caninum. All of the tested serum samples recognized N. caninum specific bands on Western blot. Low IgG serum also recognized these antigens but with lower intensity. Antibody response was also evaluated using a commercially available ELISA kit for N. caninum. PMID- 16697532 TI - Is there heterogeneity among syndromes of substance use disorder for illicit drugs? AB - The use of DSM criteria to evaluate liability to substance use disorders (SUDs) and to identify SUD phenotypes may not provide the sensitivity required to identify genes associated with vulnerability to SUDs. The purpose of this study is to evaluate a number of basic aspects of substance use that may be more proximal than full SUDs to risk genes, some of which may thus have greater potential utility as phenotypes in subsequent molecular genetic analyses. In this paper we present results from the first stage of our planned analyses, focusing on how individual symptoms of abuse and dependence may be used to create alternate phenotypes for SUDs. Specifically, we used factor analysis and biometrical modeling on each symptom of illicit substance abuse and dependence within different types of substances, and compared and contrasted factor patterns and heritabilities across the different substances. These analyses were carried out using a population-based sample of 3372 male-male twin pairs from the Vietnam Era Twin Registry who participated in the Harvard Twin Study of Substance Abuse. We obtained extensive data from these participants on substance use and SUDs via telephone interview in 1992, including data on the illicit substances: opiates, cocaine, cannabis, sedatives, stimulants, and psychedelics. The results indicate that: A) although a one-factor model assuming a single underlying liability for abuse and dependence symptoms and behaviors can be rejected for most substances, there is no uniform support for a two-factor model differentiating between abuse versus dependence; B) patterns of symptoms or behaviors reported by substance users vary across substances; C) not all symptoms or behaviors contribute equally to the presentation of an SUD; and D) the heritability of symptoms or behaviors of substance users varies both within and between substances. These results represent important first steps in facilitating the search for SUD-risk genes in subsequent high-throughput molecular genetic analyses by providing alternate phenotypes that may have both optimal validity and increased heritability. PMID- 16697533 TI - Identifying cluster subtypes for the prevention of adolescent smoking acquisition. AB - School-based smoking prevention programs are typically identical for all students. Tailoring prevention materials to focus on individual needs with an emphasis on students at highest risk is a promising alternative. Recent prevention programs have tailored materials based on the Stages of Acquisition, an extension of the Stages of Change used to tailor smoking cessation materials effectively for adults. Three stages of acquisition have been identified: Acquisition Precontemplation (aPC), Acquisition Contemplation (aC) and Acquisition Preparation (aPR). However, about 90% of nonsmoking adolescents classify themselves in the aPC stage. A cluster analysis was performed, using the Decisional Balance and Situational Temptations scales, for three random subsamples of adolescents within the aPC stage (N(1)=N(2)=N(3)=514). Four distinct subtypes were identified in each subsample: High Risk, Protected, Ambivalent, and Risk Denial. External validity was established using family support for nonsmoking, peer variables, and stage classification at follow-up assessment (12, 24, and 36 months). Family support for nonsmoking was related to subtype much more strongly than peer interactions. Subjects in the Protected subgroup were the most likely to remain in the aPC stage at each follow-up assessment. Subtype membership, along with membership in the aC and aPR stages, provides important additional information for tailoring smoking prevention materials. Tailored interventions can focus on those adolescents at highest risk and limit or avoid expending resources on those at very low risk. PMID- 16697534 TI - A common structural mechanism underlying GCMB mutations that cause hypoparathyroidism. AB - Hypoparathyroidism, either of acquired or inherited origin, is a heterogenous group of human disorders caused by a defective calcium homeostasis clinically known as hypocalcemia and hyperphosphatemia. Two mutations (R47L, G63S) in the DNA binding domain of the parathyroid-specific transcription factor GCMB have been reported to be linked to hypoparathyroidism. Both mutations cause a loss of transactivation either with (R47L) or without (G63S) a concomitant loss of DNA binding. Despite these differences with respect to their DNA binding ability, molecular modeling of the wild type and mutant GCMB-DNA complexes reveals a common regular pattern of molecular interactions which is apparently crucial for the integrity of the GCM DNA binding domain and is altered by the respective mutations. The significance of this model is substantiated from an investigation of all biochemically known mutations of the DNA binding domain of GCM proteins that impede transactivation. All of them share the proposed molecular mechanism and thus can be predicted correctly by our model. This mechanistic commonness allows the prediction of 21 additional residues of which mutation might critically affect the transactivating ability of GCMB and thus might be linked to disease when present in patients. PMID- 16697535 TI - Differential promoter usage for insulin-like growth factor-II gene in Chinese hepatocellular carcinoma with hepatitis B virus infection. AB - BACKGROUND: Human insulin-like growth factor-II (IGF-II) gene contains nine exons and four different promoters (P1-P4). Expression of the gene is elevated in the preneoplastic hepatic foci and hepatocellular carcinoma (HCC) of experimental animals and humans. To gain insight into transcriptional regulation of the gene in HCC, we analyzed the relative usage of the P1-P4 promoters and its correlation with the clinical and pathological characteristics in Chinese hepatocellular carcinoma with hepatitis B virus (HBV) infection. METHODS: P1-P4 usage levels of the gene in tumorous and matched adjacent nontumorous tissues from 23 HCC patients and 7 normal liver tissues were evaluated using a semiquantitative reverse-transcription polymerase chain reaction (RT-PCR) assay. The mutation status of p53 gene in HCC tissues was analyzed by PCR and sequencing. RESULTS: Transcripts from P1 were not detectable in 65.2% HCC tissues, and were expressed at low levels or not expressed in all nontumorous tissues compared with normals, but P2 usage levels showed no differences. P3 and P4 expression was significantly increased in most of HCC and almost all adjacent nontumorous tissues. There was a positive association of expression levels of both P3 and P4 transcripts in HCC tissues with the p53 mutation and presence of tumor embolus of portal vein, and expression of P3 were negatively related to differentiation of HCC. However, expression of both P3 and P4 was not associated with other parameters. CONCLUSIONS: Loss of P1 activity and reactivation of P3 and P4 are important characteristics in most of Chinese HCC with HBV infection, and increased IGF-II expression from P3 and P4 may play an active role in early proliferation of precancerous liver cells and hepatocarcinogenesis of these cases. Significant increase in fetal transcripts is associated with the p53 mutation and poor prognosis of the HCC patients and might serve as one of identification parameters of poor HCC prognosis. PMID- 16697536 TI - The cyclin-like uracil DNA glycosylase (UDG) of murine oocytes and its relationship to human and chimpanzee homologues. AB - Examination of electrophoretically resolved randomly generated PCR amplicons from mature murine oocytes revealed the presence of a short sequence with partial homology to a cyclin-like human uracil DNA glycosylase (UDG2), a member of an important group of base-excision enzymes that remove misincorporated or cytosine derived uracil from nascent DNA. A combination of 3' R.A.C.E. and in silico 'cloning' of the 5' region of the original EST uncovered a predicted murine mRNA sequence with limited homology to human UDG2 at both nucleotide and amino acid levels. An alternative contig for human UDG2 showed stronger similarity with the homologous murine and chimp genomic and protein sequences and contained both N and C terminal cyclin box elements. This contig is also an Ensembl annotated alternative splice-site variant of the UDG2 gene (Udg2v2). We suggest that uracil excision repair in oocytes is likely to be mediated by Udg2v2, or alternatively that Udg2v2 is involved in a process related to oocyte-specific maturation by virtue of its cyclin-like domains. PMID- 16697537 TI - Meeting the educational needs of multiple birth children. AB - There has been a significant increase in the number of twins and higher multiples so that one child in 33 is now a multiple. It is therefore not unusual for schools to have several sets of twins, as well as triplets and even higher multiples. By being the same age and in the same school year if not class, twins and higher multiples are not like brothers and sisters born closely together. Teachers and parents need to be aware of particular issues that may affect the physical, intellectual, personal, social and emotional development of multiple birth children, and to ensure that school policy and practice include this special group of children and parents. These issues include: preterm birth catch up and implications for starting school; the balance of competition and cooperation among multiples; separation in school and the evidence from recent longitudinal studies; legislative and other initiatives on the development of school policy; the particular needs of higher multiples. PMID- 16697538 TI - Electrospray encapsulation of water-soluble protein with polylactide. Effects of formulations on morphology, encapsulation efficiency and release profile of particles. AB - Bovine serum albumin (BSA)-loaded poly(lactide) (PLA) particles were prepared using an electrospraying technique, in which a sufficiently strong electric field was applied to overcome the surface tension of a droplet. A comprehensive investigation was conducted on the effects of independent variables organic/aqueous phase volume ratio and BSA/PLA weight ratio on the dependent variables viscosity, electrical conductivity, surface tension; the morphologies, sizes, and yields of particles; BSA encapsulation efficiency (EE); and in vitro release. An increase in the organic/aqueous phase ratio increased the viscosity and decreased the electrical conductivity of the emulsions, while the viscosity increased with BSA/PLA ratio. In general, spherical particles, with smooth surface and without visible pores, were observed. However, the spherical shape was lost as the organic/aqueous phase ratio decreased and the BSA/PLA ratio decreased. The particle sizes ranged from 0.84+/-0.18 to 3.95+/-0.51mum and the yield was in the range of 64.3+/-1.8 to 80.1+/-2.6%. EE of BSA was between 22.9 and 80.6%, and was increased with organic/aqueous phase ratio and decreased with increasing BSA/PLA ratio. In vitro release of BSA from the particles was reduced with increasing organic/aqueous phase ratio and was enhanced by the increase in the BSA/PLA ratio. PMID- 16697539 TI - Evaluation of melt granulation and ultrasonic spray congealing as techniques to enhance the dissolution of praziquantel. AB - Praziquantel (PZQ), an anthelminthic drug widely used in developing countries, is classified in Class II in the Biopharmaceutics Classification Systems; this means that PZQ has very low water solubility and high permeability, thus the dissolution is the absorption rate-limiting factor. The aim of this work was to evaluate the suitability of melt granulation and ultrasonic spray congealing as techniques for enhancing the dissolution rate of PZQ. Granules in high shear mixer were prepared by melt granulation, using polyethylene glycol 4000 or poloxamer 188 as meltable binders and alpha-lactose monohydrate as a filler. Quite regularly shaped granules having main size fraction in the range 200-500 microm were obtained using both formulations; however, only poloxamer 188 granules demonstrated a significant (P=0.05) increase of the PZQ dissolution rate compared to pure drug. To evaluate the potential of ultrasonic spray congealing, Gelucire 50/13 microparticles having different drug to carrier ratios (5, 10, 20 and 30%, w/w) were then prepared. The results showed that all the microparticles had a significant higher dissolution rate (P=0.05) respect to pure PZQ. The increase of the PZQ content considerably decreased the dissolution rate of the drug: 5 and 10% PZQ loaded systems evidenced dissolution significantly enhanced compared to 20 and 30% PZQ microparticles. The microparticle's characterisation, performed by Differential Scanning Calorimetry, Hot Stage Microscopy, X-ray powder diffraction and FT-Infrared analysis, evidenced the absence of both modifications of the solid state of PZQ and of significant interactions between the drug and the carrier. In conclusion, melt granulation and ultrasonic spray congealing could be proposed as solvent free, rapid and low expensive manufacturing methods to increase the in vitro dissolution rate of PZQ. PMID- 16697541 TI - Acute and subacute toxicity study of the ethanol extract from Lonicera japonica Thunb. AB - The ethanol extract from the leaves of Lonicera japonica Thunb. was evaluated for acute and subacute toxicity. The single oral dose of the ethanol extract at 5,000 mg/kg did not produce mortality or significant changes in the general behaviour and gross appearance of the internal organs of rats. In subacute toxicity study, the ethanol extract was administered orally at a dose of 1,000 mg/kg/day for a period of 14 days. The satellite group was treated with the ethanol extract at the same dose and the same period and kept for another 14 days after treatment. There were no significant differences in the body and organ weights between the control and the treated group of both sexes. Hematological analysis and clinical blood chemistry revealed no toxicity effects of the extract. Pathologically, neither gross abnormalities nor histopathological changes were observed. PMID- 16697540 TI - Prevention of hepatic injury by a traditional Chinese formulation, BJ-JN, in mice treated with Bacille-Calmette-Guerin and lipopolysaccharide. AB - The hepatoprotective effects of BJ-JN (a traditional Chinese formulation) were evaluated in Bacille-Calmette-Guerin and lipopolysaccharide (BCG/LPS)-induced immunological liver injury (ILI) in mice. BJ-JN (0.75, 1.5, 3 g/kg) was administered via gavage daily for 10 days. Liver index (liver weight/body weight), serum levels of alanine aminotransferase (ALT), hepatic nitric oxide (NO), malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, splenocyte proliferation, production of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL-1) by peritoneal macrophages, and histopathologic changes of the liver were evaluated following the 10 days treatment. BJ-JN (0.75, 1.5, 3 g/kg) effectively reduced the BCG/LPS-induced elevated liver index, serum ALT levels, hepatic NO and MDA content, and restored hepatic SOD activity in ILI mice. BJ-JN treatment also alleviated diminished splenocyte proliferation induced by concanavalin A (ConA) and repressed abnormally high levels of TNF-alpha and IL 1 from peritoneal macrophages. The histopathological analysis suggested that BJ JN reduced the degree of liver injury of ILI mice. These results suggest that BJ JN has a protective and therapeutic effect on ILI mice, which might be associated with its antioxidant properties, ability to reduce NO production and immunoregulatory function. PMID- 16697542 TI - Diverse morphological lesions and serious arrhythmias with hemodynamic insults occur in the early myocardial contusion due to blunt impact in dogs. AB - To investigate the morphology and hemodynamics of the early myocardial contusion, an animal model of cardiac contusion was established by impact to the precordial region at sternum at velocity of 10.0m/s with a mechanical elastic-cord propelled impactor in 19 dogs. The electrocardiogram and both the left and right intra ventricular pressures were recorded continuously throughout the experiment. Histological and immunohistochemical examinations of myoglobin, creatine kinase MB and fibrinogen were conducted. At the moment of impact, abrupt over-pressures within the left and right ventricles occurred with concomitant serious arrhythmias followed by variety of cardiac conduction disorders and depressed left and right ventricular systolic pressures during the observation times. Histologically, lesions of myocardial contusions were identified at subepicardial, myocardial or subendocardial layer as interstitial hemorrhage, disruption or coagulative necrosis as well as contraction band necrosis of the muscle fibers, which might be categorized into the hemorrhagic, necrotized and mixed forms. The three forms of lesions were found to exist independently, or co existed in a heart. However, severity of the lesions varied greatly with different parts even within a heart. Intravascular thromboses were occasionally discovered post-impact. Immunohistochemically, loss of myoglobin and creatine kinase-MB from cardiac cells, and accumulation of fibrinogen at the cell membranes were detected 5min post-impact. The intracellular accumulation of fibrinogen increased with extension of post-impact intervals. Our results indicate that diverse morphological lesions concomitant with hemodynamic compromise and serious, even fatal arrhythmias occur in the early myocardial contusion, and intravascular thromboses are occasionally produced, suggesting that traumatic myocardial ischemic lesion may be induced due to blunt impact to the precordial region. PMID- 16697544 TI - [Genetic polymorphisms and infections]. AB - Recent studies have shown that genetic variants, responsible for the different human response when facing an infectious risk, concerned the genes of proteins involved in either recognition of the infectious agent, in the inflammatory cascade, or in the coagulation process. For example, some studies clearly demonstrated that if a twin was affected by an infectious disease, the risk of infection by the same agent for the other twin was significantly higher in homozygote than in heterozygote twins. In Caucasians, a punctual mutation of the TLR2 cytosol was proved to block the response to bacterial lipoproteins and to some Gram positive bacteria and could be responsible for a greater susceptibility to septic shock. Several polymorphisms of the tlr4 gene have been involved in the onset of septic shock in postsurgery infection due to Gram(-) bacilli. Paradoxically, these variants seemed to protect against legionellosis. In pediatrics, polymorphisms of tlr4 were also clearly identified as risk factors for meningococcemia of severe bronchiolitis due to the respiratory syncytial virus (RSV) in children under 2 years of age. A polymorphism of the TLR5 receptor gene, which creates a stop codon and which is responsible for a nonfunctionality of the receptor was associated with the onset of severe legionellosis. Analysing the functions of these genetic polymorphisms in the onset of sepsis will open the way to a lot of research on specific treatments focused on genetic abnormalities. PMID- 16697543 TI - Serotonin turnover in different duration of sleep recovery in discrete regions of young rat brain after 24 h REM sleep deprivation. AB - Sleep plays an important role in restorative function and serotonin (5 hydroxytryptamine: 5HT) equally plays important roles in sleep. Though various studies have revealed the roles of 5HT in sleep/wake cycle, the mechanism involved is yet unclear. In the present study we investigated alteration of the 5HT turnover in various regions of the young rat brains after 24 hours (h) REM sleep (sREM) deprivation to elucidate the roles of 5HT in sleep restoration function in the these regions. The 5HT turnover was evaluated by the ratio of 5 hydroxyindole acetic acid against 5HT. The sREM deprivation was performed by the inverted flowerpot technique. The 5HT turnover showed significant alteration in the all regions of the brain examined after 24h sREM deprivation, particular depending on the brain region. These results revealed that sREM modulates the 5HT turnover in the brain with region specificity and this may be one of the restorative functions of sleep indicating that sREM is regionally generated. PMID- 16697545 TI - [Relapse of Plasmodium falciparum malaria in a patient treated with artesunate]. AB - Chemoprophylaxis and the curative treatment of malaria are well documented in France. Nevertheless this data is still not accounted for by healthcare professionals and in the global population, and no longer approved treatments may be prescribed. The authors report the case of a 24-year-old female traveler having stayed in Africa where she was used to treat fever with artesunate. Soon after her return, she presented with uncomplicated Plasmodium falciparum malaria once again treated with artesunate. The evolution was initially favorable but a relapse occurred 3 weeks later. A conventional mefloquine treatment lead to a final cure. This observation confirms that artesunate monotherapy in malaria exposes to a risk of relapse. Artesunate should not be used as monotherapy in P. falciparum malaria. PMID- 16697546 TI - Dietary fatty acids intakes and rate of mild cognitive impairment. The Italian Longitudinal Study on Aging. AB - The possible impact of diet, particularly the intake of fatty acids, on cognitive decline and dementia was addressed recently by several studies. We investigated the role of dietary fatty acids on the rate of mild cognitive impairment (MCI) in a population-based, prospective study carried out on 278 and 186 nondemented elderly subjects (65-84 years) at the 1st (1992-1993) and 2nd (1995-1996) survey from the cohort of Casamassima, Bari, Italy (n=704), one of the eight centers of the Italian Longitudinal Study on Aging. During the median follow-up of 2.6 years, 18 new events of MCI were diagnosed, and high polyunsaturated fatty acids (PUFA) intake appeared to be a protective factor against the development of MCI [hazard ratio (HR): 0.65, 95% confidence interval (CI): 0.43-0.98, trend-test, df=1, p<0.04]. However, when we controlled for the possible confounders (age, sex, education, Charlson comorbidity index, and total energy intake), the HR slightly changed, and the highly skewed 95% CI, while not statistically significant, may be important (HR: 0.62, 95% CI: 0.34-1.13, p=0.12). In our population, dietary fatty acids intakes were not associated with incident MCI in older age, only high PUFA intake evidenced a borderline nonsignificant trend for a protective effect against the development of MCI. PMID- 16697547 TI - Debulking surgery for advanced thymoma. AB - AIMS: This study was conducted to evaluate the efficacy of debulking surgery in the treatment of locally advanced but operable malignant thymoma. METHODS: We reviewed 43 cases with incompletely resected stage III and IVa malignant thymoma managed between January 1987 and December 2002. RESULTS: Twenty-two had stage III and 21 had stage IVa disease. Maximal debulking was performed in 15 patients, nine with stage III and six with stage IVa disease. Nine patients also had myasthenia gravis (MG). Using univariate Kaplan-Meier analysis, we found that maximal debulking surgery, RT, and with the presence of MG were associated with better survival. Debulking resulted in a better outcome than non-debulking surgery (mean survival: 106 months vs 57.2 months). After adjustment with multivariate analysis, RT and MG were both associated with better survival. CT did not appear to be beneficial for advanced thymoma. CONCLUSIONS: RT is independently associated with a better outcome in locally advanced thymoma. Debulking surgery apparently allows for a better response to RT. PMID- 16697548 TI - Cytotoxicity of single-wall carbon nanotubes on human fibroblasts. AB - We present a toxicological assessment of five carbon nanomaterials on human fibroblast cells in vitro. We correlate the physico-chemical characteristics of these nanomaterials to their toxic effect per se, i.e. excluding catalytic transition metals. Cell survival and attachment assays were evaluated with different concentrations of refined: (i) single-wall carbon nanotubes (SWCNTs), (ii) active carbon, (iii) carbon black, (iv) multi-wall carbon nanotubes, and (v) carbon graphite. The refined nanomaterial that introduced the strongest toxic effect was subsequently compared to its unrefined version. We therefore covered a wide range of variables, such as: physical dimensions, surface areas, dosages, aspect ratios and surface chemistry. Our results are twofold. Firstly, we found that surface area is the variable that best predicts the potential toxicity of these refined carbon nanomaterials, in which SWCNTs induced the strongest cellular apoptosis/necrosis. Secondly, we found that refined SWCNTs are more toxic than its unrefined counterpart. For comparable small surface areas, dispersed carbon nanomaterials due to a change in surface chemistry, are seen to pose morphological changes and cell detachment, and thereupon apoptosis/necrosis. Finally, we propose a mechanism of action that elucidates the higher toxicity of dispersed, hydrophobic nanomaterials of small surface area. PMID- 16697549 TI - Prevention and recovery of (mu(3)-diethylentriamino)-chloro-palladium(II) chloride induced inhibition of Na/K-ATPase by SH containing ligands--L-cysteine and glutathione. AB - The effect of (mu(3)-diethylentriamino)-chloro-palladium(II)-chloride ([PdCl(dien)]Cl) on the activity of Na/K-ATPase from porcine cerebral cortex was studied in vitro, in the absence and presence of -SH containing ligands L cysteine and glutathione (GSH). The aim of the study was to elucidate the mechanism of [PdCl(dien)](+) induced inhibition of the enzyme activity and to examine the ability of thiols to prevent and recover the inhibition. The coordinative interaction between [PdCl(dien)](+) and enzyme was verified by UV and (1)H NMR spectra. The semblance in the changes in absorption spectra of [PdCl(dien)](+) in the presence of Na/K-ATPase and thiols (L-cysteine and GSH) suggested that the complex ion interacts with enzymatic sulfhydryl groups. [PdCl(dien)](+) inhibited the enzyme activity in a dose-dependent manner. The Hill analysis of the inhibition curve yielded the half-maximum inhibitory activity value, IC(50)=1.21 x 10(-4)M, and Hill coefficient, n=0.7, suggesting the negative cooperation for binding of [PdCl(dien)](+) to the enzyme. Dependence of the initial reaction rate on the concentration of MgATP(2-) exhibited typical Michelis-Menten kinetics in the absence and presence of the inhibitor. Kinetic analysis showed that [PdCl(dien)](+) inhibited Na/K-ATPase by reducing the maximum reaction rate (V(max)), rather than changing the affinity to the substrate (K(m)). Kinetic parameters derived using Lineweaver-Burk transformation of experimental data indicated the non-competitive nature of Na/K-ATPase inhibition. The inhibitory constant, K(i)=1.05 x 10(-4)M, was determined from secondary replot of Lineweaver-Burk graph, and correlated with stability constants of [Pd(dien)(thiol)] complexes. 1 x 10(-3)M L-cysteine or GSH prevented the enzyme inhibition induced by Pd(II) complex cation when present below 1 x 10( 4)M. The both thiols completely reversed the inhibited activity in the concentration dependent manner, due to the complex formation with [PdCl(dien)](+). PMID- 16697550 TI - Subjective versus objective behavioral ratings following two analogue tasks: a comparison of socially phobic and non-anxious adolescents. AB - Although results regarding objective social skills deficits in adults are inconsistent, research with youth have consistently demonstrated such deficits. Furthermore, research has consistently demonstrated subjective appraisal of social skill deficit in both youth and adults with social phobia or social anxiety. As a result, research has begun to examine the presence of a negative perceptual bias in individuals with social phobia and social anxiety. The purpose of the present study was to extend the findings of social skill deficits to an adolescent sample by investigating differences between adolescents with social phobia and non-anxious peers with regard to objective and subjective behavioral ratings. In addition, the presence of a negative perceptual bias was investigated by examining discrepancy scores between these ratings. The results of the current study provide support for the presence of social skill deficits in a socially phobic adolescent population and are the first to suggest that a limited negative bias exists in this population. PMID- 16697551 TI - CC chemokines in zebrafish: evidence for extensive intrachromosomal gene duplications. AB - Chemokines are a family of structurally related chemotactic cytokines that regulate the migration of leukocytes. CC chemokines represent the largest subfamily of chemokines, with 28 genes in mammals. In recent studies in channel catfish, Ictalurus punctatus, we identified 26 distinct CC chemokine transcripts and obtained the genomic sequences and structures of 23 CC chemokine genes. However, without the availability of similar sets of CC chemokines in closely related species or a sequenced genome in catfish, it was difficult to make inferences as to the origins and modes of duplication of these molecules or to analyze conserved synteny between teleost and mammalian CC chemokines. Here, we have identified as many as 46 loci in the zebrafish genome that encode putative CC chemokines. The zebrafish CC chemokines are highly clustered on several chromosomes and show evidence of extensive, species-specific intrachromosomal duplications. PMID- 16697552 TI - A second-generation genetic linkage map of the baboon (Papio hamadryas) genome. AB - Construction of genetic linkage maps for nonhuman primate species provides information and tools that are useful for comparative analysis of chromosome structure and evolution and facilitates comparative analysis of meiotic recombination mechanisms. Most importantly, nonhuman primate genome linkage maps provide the means to conduct whole genome linkage screens for localization and identification of quantitative trait loci that influence phenotypic variation in primate models of common complex human diseases such as atherosclerosis, hypertension, and diabetes. In this study we improved a previously published baboon whole genome linkage map by adding more loci. New loci were added in chromosomal regions that did not have sufficient marker density in the initial map. Relatively low heterozygosity loci from the original map were replaced with higher heterozygosity loci. We report in detail on baboon chromosomes 5, 12, and 18 for which the linkage maps are now substantially improved due to addition of new informative markers. PMID- 16697553 TI - A mouse TRAPP-related protein is involved in pigmentation. AB - We identified a new spontaneous recessive mutation in the mouse, mhyp (mosaic hypopigmentation), in a screen for novel proviral integration sites in a multiple ecotropic provirus mapping stock. Integration of an 8.4-kb retrovirus results in mosaic loss of coat pigment in mhyp homozygotes. Patchy loss of pigmentation in the retinal pigmented epithelial layer of the eye with abnormal melanosomes is also evident. We mapped mhyp to mouse chromosome 7 and cloned the underlying gene. mhyp is a defect in the Trappc6a gene. Expression of Trappc6a is markedly diminished in mhyp homozygotes. The normal protein, TRAPPC6A, is a subunit of the TRAPP (transport protein particle) I and II complexes. While TRAPP complexes are essential for ER-to-Golgi and intra-Golgi vesicle trafficking in yeast, TRAPP subunits participate in additional, including post-Golgi, transport events in mammals. The data implicate mammalian TRAPPC6A in vesicle trafficking during melanosome biogenesis. PMID- 16697555 TI - Botryoid odontogenic cyst versus glandular odontogenic cyst. PMID- 16697554 TI - Manganese acts centrally to activate reproductive hormone secretion and pubertal development in male rats. AB - Manganese (Mn) is an important element for normal growth and reproduction. Because Mn accumulates in the hypothalamus and is capable of stimulating puberty related hormones in female rats, we assessed whether this metal could cause similar effects in male rats. We have demonstrated that MnCl2, when administered acutely into the third ventricle of the brain, acts dose dependently to stimulate luteinizing hormone (LH) release. Furthermore, there was a dose dependent stimulation in the secretion of LH-releasing hormone (LHRH) from the medial basal hypothalamus in vitro, and administration of an LHRH receptor antagonist in vivo blocks Mn-induced LH release. To assess potential chronic effects of the metal, male pups were supplemented with 10 or 25 mg MnCl2 per kg by gastric gavage from day 15 until days 48 or 55, at which times developmental signs of spermatogenesis were assessed. Results demonstrate that while significant effects were not observed with the 10 mg/kg dose, the animals receiving the 25 mg/kg dose showed increased LH (p<0.05), FSH (p<0.01) and testosterone (p<0.01) levels at 55 days of age. Furthermore, there was a concomitant increase in both daily sperm production (p<0.05) and efficiency of spermatogenesis (p<0.05), demonstrating a Mn-induced acceleration in spermatogenesis. Our results suggest Mn is a stimulator of prepubertal LHRH/LH secretion and may facilitate the normal onset of male puberty. These data also suggest that the metal may contribute to male precocious pubertal development should an individual be exposed to low but elevated levels of Mn too early in life. PMID- 16697557 TI - Novel plasmid-mediated CTX-M-8 subgroup extended-spectrum beta-lactamase (CTX-M 40) isolated in the UK. PMID- 16697558 TI - Evaluation of a cefoxitin disk diffusion test for the detection of mecA-positive methicillin-resistant Staphylococcus saprophyticus. AB - In order to validate the current Clinical and Laboratory Standards Institute (CLSI) criteria for the detection of mecA-mediated resistance in Staphylococcus saprophyticus, 101 clinical isolates, including 8 mecA-positive isolates, were investigated. All the isolates were in the range of the resistant category for coagulase-negative staphylococci with the 1 microg oxacillin disk diffusion method and agar dilution method, despite 93 isolates (92%) being mecA-negative. On the other hand, the 30 microg cefoxitin disk diffusion method showed clearly distinguishable zone diameters between the mecA-positive and -negative isolates. However, four of the mecA-negative isolates that would be considered resistant were false positive, and the current interpretive criteria of the CLSI may thus require reconsideration. This study suggests that the cefoxitin disk diffusion method could be more suitable than the oxacillin disk diffusion method for detecting mecA-mediated resistance in S. saprophyticus. PMID- 16697559 TI - Results of an interlaboratory test on antimicrobial susceptibility testing of bacteria from animals by broth microdilution. AB - A standard operating procedure for the determination of minimum inhibitory concentrations (MICs) of antimicrobial agents by the broth microdilution method was developed and evaluated for its fitness for use in an interlaboratory ring trial involving 46 routine diagnostic laboratories. All laboratories tested five strains (one reference strain and four field strains) against a total of 22 different antimicrobial agents. Gram-negative strains were tested against 16 different antimicrobial agents and Gram-positive strains against 14 different antimicrobial agents. Tests were performed once a week for three consecutive weeks. At least 80% of the results determined by 35 of the 46 participating laboratories were within the expected range (mode MIC+/-1 dilution step), with the 18 participating laboratories experienced in MIC determination showing a slightly higher mean percentage of accurate results (89.3% reproducible results) than the 28 non-experienced laboratories (86.7% reproducible results). The most accurate results were obtained for the Escherichia coli field strain, whilst the results for the Streptococcus uberis field strain showed the highest error rate. Among the 22 antimicrobial agents tested, the highest variabilities in the results (mean value for all antimicrobial agents 12.3%) were recorded for ceftiofur (27.8%), penicillin G (20.8%) and cefoperazone (20.6%). PMID- 16697560 TI - Resistance of different Chlamydia-like organisms to quinolones and mutations in the quinoline resistance-determining region of the DNA gyrase A- and topoisomerase-encoding genes. AB - Parachlamydia acanthamoebae has been shown to be resistant to fluoroquinolones. In this study, we determined the susceptibility of Neochlamydia hartmannellae and Simkania negevensis to quinolones using amoebal co-culture in the presence of serial dilutions of ciprofloxacin. The gyrA and parC quinoline resistance determining regions (QRDRs) were amplified and sequenced, and amino acid substitutions that may explain the observed quinolone resistance were investigated. Neochlamydia and Simkania were resistant to 16 microg/mL ciprofloxacin. Substitution at positions 83 and 70 of GyrA QRDR were present in Neochlamydia and in both Parachlamydia strains studied. For Simkania, substitutions at positions 99 and 101 in the GyrA QRDR and at positions 81 and 84 in the ParC QRDR were observed. Resistance of Parachlamydia, Neochlamydia and Simkania to quinolones is likely due to these mutations. PMID- 16697561 TI - Drug permeability across a phospholipid vesicle-based barrier 2. Characterization of barrier structure, storage stability and stability towards pH changes. AB - Recently we reported on the development of a phospholipid vesicle-based barrier as a medium throughput method for screening of drug permeability. The aim of this present study is to characterize the barrier structure, including an estimation of the amount of phospholipid within it, its storage stability and its stability over various pH ranges found in different parts of the gastrointestinal tract. The amount of lipid in the barrier was quantified using a colorimetric phospholipase D-based assay. The total amount averaged 3.30mg phospholipid per barrier. The preparation process comprises the consecutive deposition of two types of liposomes on a filter support. We estimated that the smallest liposomes, with a mean diameter of 298nm, would fill the pore volume of the filter when tightly packed. The volume of the bigger liposomes, deposited on top of the filter, was calculated to generate a 0.1mm thick layer. Visualisation of fluorescently labelled liposomes by confocal laser-scanning microscopy confirmed that the pores of the filter were completely filled with liposomes and that there was a liposome layer on top. Small angle X-ray scattering (SAXS) analysis was used to study the lamellarity of the liposomes. The liposomes contained oligo- and/or multilamellar structures before and after deposition. The functionality of the barriers during storage at three different temperatures was examined for a period of up to 4 weeks by measuring the permeability of the hydrophilic marker calcein across them. The conclusion was that the phospholipid vesicle-based barriers could be stored at -80 degrees Celsius for up to 2 weeks without significant changes. The stability of the barriers in a pH range from 2.0 to 8.0 was investigated by performing permeation studies with fluorescein at different pH values. It was found that the phospholipid vesicle-based barrier did not lose its integrity within this range. Thus, the barriers appear suitable for further studies to provide insight into segmental absorption in the human gastrointestinal tract. Furthermore, because the phospholipid vesicle-based barrier can be stored, larger batches can be produced. This makes the phospholipid vesicle-based barrier more appropriate for high throughput screening. PMID- 16697562 TI - [Mechanisms of action of general anesthetics]. PMID- 16697563 TI - [Improving communication between the radiologist, the patient and his parents]. PMID- 16697564 TI - [Preoperative assessment of the child with congenital heart disease]. PMID- 16697565 TI - [Inherited renal diseases and prenatal diagnosis]. PMID- 16697566 TI - [Medicoeconomic results of the pediatric unit at Le Havre hospital]. PMID- 16697567 TI - [The place of the parents in the outpatient investigation of adolescents by pediatricians]. PMID- 16697568 TI - [Guidelines for improving the screening of developmental dysplasia of the hip]. PMID- 16697569 TI - [Health cost and imaging]. PMID- 16697570 TI - Developing adolescent services--crossing the divide. PMID- 16697571 TI - [Imaging of neonatal brain disorders: doubts and certitudes]. PMID- 16697572 TI - [Management of acute hypophosphatemia]. PMID- 16697573 TI - [Proximal tubular impairment (Fanconi syndrome)]. PMID- 16697574 TI - [Outcome after non-accidental head injury in children]. PMID- 16697575 TI - [Pediatric prediction models for evaluation of bone mass with DEXA]. PMID- 16697576 TI - [Hip ultrasonography. Need for quality examination]. PMID- 16697577 TI - [Back pain in children]. PMID- 16697579 TI - [French legislation and radiation dose control in paediatric computed tomography]. PMID- 16697578 TI - [A painful mass of the thigh in a child]. PMID- 16697580 TI - [Sedation in children: how and for who?]. PMID- 16697581 TI - [Do anesthetics impair brain development?]. PMID- 16697582 TI - [Gastroesophageal reflux related otolaryngologic diseases]. PMID- 16697583 TI - [Dynamic ultrasonographic study of hip articular stability]. PMID- 16697584 TI - [Guidelines for diagnostic imaging in children and responsibility of the referring physician]. PMID- 16697585 TI - [Immunization in France: high quality regulation]. PMID- 16697586 TI - [Spasticity management and progress in ambulatory cerebral palsy]. PMID- 16697588 TI - [Growth and limb pains]. PMID- 16697587 TI - [Hospital network in pediatric oncology]. PMID- 16697589 TI - [Abdominal pain in children. Traps and diagnosis]. PMID- 16697590 TI - [Immunization: adverse event reporting system]. PMID- 16697591 TI - [The radiologist and the discovery of a cerebral abnormality in the fetus]. PMID- 16697592 TI - [Long term central venous catheters: indications and technique]. PMID- 16697593 TI - [Kidney failure in the newborn]. PMID- 16697594 TI - [Guidelines for diagnostic imaging in emergency care]. PMID- 16697595 TI - [Anesthesia for patent ductus arteriosus closure]. PMID- 16697596 TI - [Failure in the screening and diagnosis of developmental dysplasia of the hip]. PMID- 16697597 TI - [Henoch-Schonlein purpura]. PMID- 16697598 TI - [Morphological hip examination by ultrasonography according to Graf]. PMID- 16697599 TI - [Management of childhood obesity]. PMID- 16697600 TI - [Hypnosis, child and pain]. PMID- 16697601 TI - [Turbulence in emergency ward. Agitation of children and adolescents]. PMID- 16697602 TI - [Brain MR imaging in shaken-baby syndrome: how? when and why?]. PMID- 16697603 TI - [Malignant bone tumours in children and adolescent]. PMID- 16697604 TI - [Renal involvement in systemic diseases]. PMID- 16697605 TI - [Management of long lasting vascular access]. PMID- 16697606 TI - [Network for investigation of medicinal products in children]. PMID- 16697607 TI - [Central venous catheter: epidemiology, diagnosis, prevention and treatment of infectious complications]. PMID- 16697608 TI - [The use of antidepressants to treat depression in children and adolescents]. PMID- 16697609 TI - [A virtual unit: experience from the institut Curie]. PMID- 16697610 TI - [MRI morphometry, an insight into brain function]. PMID- 16697611 TI - [Adolescents with cancer: a specific population]. PMID- 16697612 TI - [Growth hormone and bone]. PMID- 16697613 TI - [The unit for adolescent with cancer, institut Gustave-Roussy]. PMID- 16697614 TI - [Isolated health professionals in paediatric palliative care]. PMID- 16697615 TI - [Apparent malformations in an African area: factors influencing consultation delay in pediatric surgery]. AB - AIM: To analyze the attitudes of the Guinean and of the Ivory Coast communities leading to delayed consultation despite apparent malformations in children. PATIENTS AND METHODS: From January 1, 2000 to December 31, 2002, we carried out a prospective investigation in the paediatric surgery units of the Donka teaching hospital (Conakry) and Cocody, Treichville and Yopougon (Abidjan) teaching hospital. One hundred and two children affected with apparent malformations were included. The studied variables were: age, sex, ethnos group, religion, socio economic level and the cultural designs of the families. RESULTS: Sex ratio male/female was 1,5 and the average age at first consultation was 17 months. Seventy-six per cent of the children carrying apparent malformations at birth were seen at an age ranging from 1 to 30 months. Orthopaedic malformations were prominent (44%) and led especially to negative reactions of the entourage of the patients. Some religious beliefs took a part of the delayed consultation and impaired relationships between the 2 parents. The low socio-economic level (54%) was determining in the delayed consultation. The birth of a child with malformation in the malinke, akan krou community could be understood like a parchment from a god or a witchcraft. CONCLUSION: The contributive factors of the delay to the consultation of the children carrying apparent malformations in the communities Guinean and of the Ivory Coast are poverty, ignorance and some religious beliefs. Education and well understanding of these reasons in developing country should improve the acceptance and taking care of these children as well as the development of medical insurance system. PMID- 16697616 TI - [Nutritional evaluation in neonatology: which place for Dual Energy X Ray Absorbtiometry (DEXA)?]. PMID- 16697617 TI - [Back pain in children]. PMID- 16697618 TI - [Low radiation doses in children, benefit risk approach]. PMID- 16697619 TI - [Pelvic calcifications in a female infant]. PMID- 16697620 TI - [Gluten-free diet: a challenging follow-up]. PMID- 16697621 TI - [Prospective comparison of child asthma education in the emergency department and at scheduled follow-up consultation]. AB - OBJECTIVES: To assess asthma control in asthmatic children attending the emergency department and to compare it with that for children with scheduled specialist follow-up. POPULATION AND METHODS: Between September 2002 and September 2003, we included asthmatic children aged 6 to 16 years, attending the emergency department (group U) or pulmonology follow-up appointments (group C) at Necker Hospital, Paris, France. We used the Asthma Therapy Assessment Questionnaire (ATAQ), completed by the parents and children aged 10 years and over. RESULTS: We interviewed 156 families and included 144 (92%) in the final analysis (77 in group U). The 2 groups did not differ in age, sex ratio, age at onset and asthma diagnosis and the frequency of asthma attacks. Children in group U received less daily maintenance treatment (P<0.01) but reported more severe asthma (P<0.05). They had also experienced fewer investigations for their asthma (fewer lung function tests P=0.01 and allergy tests P=0.001). The children in group C had better controlled asthma, as assessed by both the children themselves and their parents. They also had fewer problems relating to the behavior, communication and treatment control domains. The assessment of children and parents were similar in all areas except communication. The children considered their communication problems to be more serious than their parents did. Age related differences were observed, with older children's parents having more difficulties in all domains investigated. CONCLUSION: Clinicians should consider assessing asthma control in children attending the emergency department. There is clearly an opportunity to improve the quality of asthma education for these children. Clinicians could provide children with a written plan of action for asthma attacks and information about daily self-management. Improving communication between children, their parents and doctors should be considered a key educational project. PMID- 16697622 TI - [Air pollution and asthma in children]. AB - The prevalence of asthma and allergic diseases has increased world-wide during the last quarter of the 20th century, particularly among children and adolescents. No change common to all sites where asthma has increased throughout the world has been identified, suggesting that this 'epidemic' phenomenon is likely due to multiple factors. The following have been most discussed: exposure to indoor and outdoor allergens, modification of the patterns of respiratory infections, decreasing trends of physical activity, evolution in the make-up of environmental irritants, including tobacco smoke and urban air toxicants. In this review, we point out the role of exposure to air pollutants, in addition to and in combination with other asthma enhancers or precipitators. Whereas concentrations of the 'classical' air quality indicators (SO2, CO) have more or less steadily decreased, asthma prevalence augmented in developed countries during the same period. However, the nature of the air pollution mix has deeply evolved, and should also be considered. Ambient air concentrations of industrial and house heating combustion sources of pollutants in the city have substantially decreased, but by contrast the concentrations of various ultrafine particles have increased. Now, there is in vitro and in vivo evidence that exposure to urban air particles, and particularly to diesel exhausts, elicits chronic oxidative stress and repeated inflammatory responses, so that they may enhance allergic inflammation and airway hyper-responsiveness. Several epidemiological studies suggested an association between traffic density close to places of children's residence and prevalence of respiratory symptoms, and more specifically of asthma or allergic rhinitis symptoms in them. Chronic exposure during infancy to traffic related pollutants may accelerate or even provoke, among genetically sensitive subjects, disruption of the normal regulatory and repair processes eventually contributing to the increase of asthma incidence. PMID- 16697623 TI - [Grown up congenital heart disease, current management]. PMID- 16697624 TI - [Aging with chronic renal failure from childhood]. PMID- 16697625 TI - [Back's pain, lack of words: about body's image]. PMID- 16697626 TI - [Genetics of hypophosphosphatemia]. PMID- 16697627 TI - [Hormonal effects and bone mass regulation]. PMID- 16697628 TI - [A child psychiatrist's approach]. PMID- 16697629 TI - [Which prevention for dental health?]. PMID- 16697630 TI - PCR-identification of Dunaliella salina (Volvocales, Chlorophyta) and its growth characteristics. AB - The saline pond microalga, Dunaliella salina (Dunal) Teod. maintained in De Walne's (basal) medium under laboratory conditions was confirmed by amplifying the chromosomal DNA of the microalga by PCR with specific primers MA1 and MA2. Seaweed extracts obtained from Sargassum wightii and Ulva lactuca were amended separately at 1.0%, 1.5%, 2.0% and 2.5% levels to the basal medium in order to assess their potential on the growth and concentration of pigments, viz. Chl a, Chl b and beta-carotene of the alga. beta-Carotene was isolated and visible absorption spectrum was taken at 443 and 475 nm confirmed the presence of 9-cis beta-carotene and all-trans-beta-carotene isomers. Maximum yield, highest division rate (mu) and highest pigment concentrations were observed in the cells grown in 1.5% S. wightii and 2.0% U. lactuca amended medium and these cells were subjected to DAPI staining. The results of epifluorescence microscopy and image analysis revealed a significant enhancement of the cell and nuclear area of the microalgae. PMID- 16697631 TI - Ambient pH signaling regulates expression of the serine protease gene (spr1) in pine wilt nematode-trapping fungus, Monacrosporium megalosporum. AB - We have cloned and characterized spr1, a putative serine protease gene, from a nematode-trapping fungus, Monacrosporium megalosporum. The gene was present as a single copy in the genome. The predicted protein sequence of spr1 is homologous to the putative cuticle-degrading serine proteases PII and Azo1 from the nematode trapping fungus, Arthrobotrys oligospora. In the 5' untranslated region near the initiation codon, consensus sequences to an AreA binding site, a well-known mediator of nitrogen metabolite repression in the fungus Aspergillus nidulans, a CreA binding site, a carbon response regulator in A. nidulans, and a PacC binding site, a transcription factor that responds to ambient pH signals in A. nidulans were found. However, spr1 was not regulated by carbon or nitrogen source, and exogenous protein did not induce expression of spr1. The transcription of the spr1 gene of this fungus was significantly affected by ambient pH. Based on RT PCR, the product of the spr1 gene was not transcribed at pH 4, whereas under alkaline conditions such as pH 8 and 9, the spr1 gene was transcribed well. These results indicate that the spr1 gene is controlled only by a PacC homologue. Moreover, the expression profile of the spr1 gene corresponded with the pH dependent physiology of this fungus. PMID- 16697632 TI - Turkish freshwater and marine macrophyte extracts show in vitro antiprotozoal activity and inhibit FabI, a key enzyme of Plasmodium falciparum fatty acid biosynthesis. AB - The ethanolic extracts of a number of Turkish freshwater macrophytes (Potamogeton perfoliatus, Ranunculus tricophyllus and Cladophora glomerata) and marine macroalgae (Dictyota dichotoma, Halopteris scoparia, Posidonia oceanica, Scinaia furcellata, Sargassum natans and Ulva lactuca) were assayed for their in vitro antiprotozoal activity. Trypanosoma brucei rhodesiense, Trypanosoma cruzi, Leishmania donovani and Plasmodium falciparum were used as test organisms. The cytotoxicity of the extracts was also assessed against primary rat skeletal myoblasts (L6 cells). Whereas none of the extracts were active against T. cruzi, all crude extracts displayed appreciable trypanocidal activity against T. brucei rhodesiense, with S. natans being the most active one (IC(50) 7.4microg/ml). Except for the marine alga H. scoparia, all extracts also possessed leishmanicidal potential. The best antileishmanial activity was exerted by U. lactuca and P. oceanica (IC(50)'s 5.9 and 8.0microg/ml, respectively). Five extracts that demonstrated inhibitory activity towards P. falciparum (IC(50)'s 18.1-48.8microg/ml) were simultaneously assayed against FabI, a crucial enzyme of the fatty acid system of P. falciparum, to find out whether FabI was their target. The extracts of C. glomerata and U. lactuca efficiently inhibited the FabI enzyme with IC(50) values of 1.0 and 4.0microg/ml, respectively. None of the extracts were cytotoxic towards mammalian L6 cells. This work reports for the first time antiprotozoal activity of some Turkish marine and freshwater algae, as well as a target-based antiplasmodial screening for the identification of P. falciparum FabI inhibitors from aquatic and marine macrophytes. PMID- 16697633 TI - Sensing capability of the localized surface plasmon resonance of gold nanorods. AB - We demonstrate the feasibility of using the longitudinal component of gold nanorod's surface plasmon resonance in biomolecular sensing. The sensitive dependence of the absorption maximum on the dielectric constant of the particle interfacial region makes gold nanorods a promise for constructing a biomolecular sensing scheme. The sensor containing gold nanorods, with a mean aspect ratio of 5.2, exhibits a sensitivity of ca. 366 nm/RIU (refractive index unit), which increases accordingly with the increase of the particle mean aspect ratios. Such a biosensor was further modified to demonstrate its effectiveness in quantitative detection for selective binding events, such as biotin/streptavidin pairs, through a process in which biotin molecules were chemically attached to the gold nanorods' surface prior to detection measurements. Results showed that the spectral lambda(max) shifts linearly to the concentrations of the streptavidin. The results from both experiment and model calculations strongly indicate the efficacy of the longitudinal surface plasmon absorption band in biosensing. PMID- 16697634 TI - WITHDRAWN: A radio frequency biosensor with gold nanoparticle probes. 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- 16697635 TI - Lytic phage as a specific and selective probe for detection of Staphylococcus aureus--A surface plasmon resonance spectroscopic study. AB - Rapid and reliable detection of harmful pathogens at low levels are vital due to the related environmental and economical impact. While antibodies (monoclonal or polyclonal) are successfully employed in many immunoanalysis procedures as a biorecognition element, many of them remain costly with a comparatively short shelf life and uncertain manufacturability. Additionally, they suffer from several limitations, such as susceptibility to hostile environmental stresses such as temperature, pH, ionic strength, and cross-reactivity. The development of easy available, sensitive, and robust alternative molecular recognition elements, capable of providing a very high level of selectivity are very attractive to industry and may benefit in multiple areas. Several attempts have been made to utilize fluorescent-tagged bacteriophages and phage-displayed peptides for bacterial detection. However, involvement of complex labeling and detecting procedures make these approaches time-consuming and complicated. Here, we are reporting for the first time, the label-free detection of Staphylococcus aureus using lytic phage as highly specific and selective biorecognition element and surface plasmon resonance-based SPREETA sensor as a detection platform. Lytic phage was immobilized on the gold surface of SPREETA sensor via trouble-free direct physical adsorption. The detection limit was found to be 10(4) cfu/ml. Detection specificity was investigated by an inhibition assay while selectivity was examined with Salmonella typhimurium. The preliminary results using lytic phage as a probe for bacterial detection, in combination with SPR platform are promising and hence can be employed for rapid and label-free detection of different bacterial pathogens. PMID- 16697636 TI - Gonadotropin-releasing hormone signaling in behavioral plasticity. AB - Sex and reproduction sculpt brain and behavior throughout life and evolution. In vertebrates, gonadotropin-releasing hormone (GnRH) is essential to these processes. Recent advances have uncovered novel regulatory mechanisms in GnRH signaling, such as the initiation of sexual maturation by kisspeptins. Yet despite our increasing molecular knowledge, we know very little about environmental influences on GnRH signaling and reproductive behavior. Alternative model systems have been crucial for understanding the plasticity of GnRH effects within an organismal context. For instance, GnRH signaling is under the control of seasonal cues in songbirds, whereas social signals regulate GnRH in cichlid fishes, with crucial consequences for reproduction and behavior. Analyzing cellular signaling cascades within an organismic context is essential for an integrative understanding of GnRH function. PMID- 16697637 TI - The role and expression of the protocadherin-alpha clusters in the CNS. AB - The clustered protocadherins comprise the largest subfamily of the cadherin superfamily and are predominantly expressed in the nervous system. The family of clustered protocadherins (clustered Pcdh family) is substructured into three distinct gene arrays in mammals: Pcdh-alpha, Pcdh-beta, and Pcdh-gamma. These are regulated by multiple promoters and cis-alternative splicing without DNA recombination. Pcdh-alpha proteins interact with beta1-integrin to promote cell adhesion. They also form oligomers with Pcdh-gamma proteins at the same membrane sites. During neuronal maturation, Pcdh-alpha expression is dramatically downregulated by myelination. The clustered Pcdh family has multiple variable exons that differ somewhat in number and sequence across vertebrate species. At the single-cell level, Pcdh-alpha mRNAs are regulated monoallelically, resulting in the combinatorial expression of distinct variable exons from each allele. These findings support the idea that diversified Pcdh molecules contribute to neural circuit development and provide individual cells with their specific identity. PMID- 16697638 TI - Neural stem cells as novel cancer therapeutic vehicles. AB - The startling resemblance of many of the behaviours of brain tumours to the intrinsic properties of the neural stem/progenitor cell has triggered a recent dual interest in arming stem cells to track and help eradicate tumours and in viewing stem cell biology as somehow integral to the emergence and/or propagation of the neoplasm itself. These aspects are reviewed and discussed here. PMID- 16697639 TI - Chemical structures and biological activities of rhamnolipids produced by Pseudomonas aeruginosa B189 isolated from milk factory waste. AB - The aim of this work was to study chemical structures and biological activities of rhamnolipids produced by Pseudomonas aeruginosa B189 isolated from milk factory waste. The culture produced two biosurfactants, a and b, which showed strong activity and were identified as L-rhamnopyranosyl-L-rhamnopyranosyl-beta hydroxydecanoyl-beta-hydroxydecanoate or Rha-Rha C10-C10 and L-rhamnopyranosyl-L rhamnopyranosyl-beta-hydroxydecanoyl-beta-hydroxydodecanoate or Rha-Rha C(10) C(12), respectively. Both compounds exhibited higher surfactant activities tested by the drop collapse test than several artificial surfactants such as SDS and Tween 80. Rhamnolipid a showed significant antiproliferative activity against human breast cancer cell line (MCF-7) at minimum inhibitory concentration (MIC) at 6.25 microg/mL while rhamnolipid b showed MIC against insect cell line C6/36 at 50 microg/mL. PMID- 16697640 TI - Nepsilon-thioacetyl-lysine: a multi-facet functional probe for enzymatic protein lysine Nepsilon-deacetylation. AB - Peptides containing Nepsilon-thioacetyl-lysine and Nepsilon-acetyl-lysine were evaluated for their de(thio)acetylation catalyzed by human HDAC8 and SIRT1, two distinct protein deacetylases. Nepsilon-Thioacetyl-lysine was found to be a mimic of Nepsilon-acetyl-lysine for HDAC8-catalyzed reaction, but to confer inhibition against SIRT1. These results point to further diverse applications based on Nepsilon-thioacetyl-lysine. PMID- 16697641 TI - Dipeptide vinyl sultams: synthesis via the Wittig-Horner reaction and activity against papain, falcipain-2 and Plasmodium falciparum. AB - The synthesis of phosphonate derivatives of N-phenyl- and N-benzyl-gamma- and delta-sultams, and their application in the Wittig-Horner reaction with N-Boc-L phenylalanine aldehyde to afford E- and Z-isomers, are described. These compounds were further processed to provide five dipeptide vinyl sultams, which were found to be inactive against papain at concentrations up to 50 microM. In contrast, vinyl sultams demonstrated weak activity against recombinant falcipain-2 and Plasmodium falciparum W2. PMID- 16697643 TI - A structural equation model relating impaired sensorimotor function, fear of falling and gait patterns in older people. AB - Many falls in older people occur while walking, however the mechanisms responsible for gait instability are poorly understood. Therefore, the aim of this study was to develop a plausible model describing the relationships between impaired sensorimotor function, fear of falling and gait patterns in older people. Temporo-spatial gait parameters and acceleration patterns of the head and pelvis were obtained from 100 community-dwelling older people aged between 75 and 93 years while walking on an irregular walkway. A theoretical model was developed to explain the relationships between these variables, assuming that head stability is a primary output of the postural control system when walking. This model was then tested using structural equation modeling, a statistical technique which enables the testing of a set of regression equations simultaneously. The structural equation model indicated that: (i) reduced step length has a significant direct and indirect association with reduced head stability; (ii) impaired sensorimotor function is significantly associated with reduced head stability, but this effect is largely indirect, mediated by reduced step length, and; (iii) fear of falling is significantly associated with reduced step length, but has little direct influence on head stability. These findings provide useful insights into the possible mechanisms underlying gait characteristics and risk of falling in older people. Particularly important is the indication that fear related step length shortening may be maladaptive. PMID- 16697642 TI - Notch signaling in the regulation of tumor angiogenesis. AB - The Notch signaling pathway is conserved in vertebrates and invertebrates and is involved in many developmental processes. Notch receptors and ligands are expressed on the cell surface enabling interactions between adjacent cells upon receptor-ligand binding. Notch signaling molecules have an important well documented role in vascular development, differentiation, proliferation, apoptosis and tumorigenesis. Recently, several groups have identified the importance of Notch signaling in tumor angiogenesis. Notch activity increases specifically in tumor endothelium and in various tumors types and, in some studies, Notch signaling suppresses angiogenic processes. Because the Notch signaling pathway can mediate communication between various cell types in the tumor microenvironment, interactions between tumor cells and endothelial cells might promote angiogenesis, therefore targeting the Notch pathway might provide a novel strategy for anti-angiogenic therapies. Here, we discuss recent insights of Notch signaling in tumor angiogenesis. PMID- 16697644 TI - Inconspicuous visual field defect in anterior choroidal artery territory infarction. AB - We describe a patient with a right anterior choroidal artery territory infarction and an inconspicuous left visual defect. The anterior choroidal artery is a unique artery of the cerebral circulation. The occlusion of this artery can result in dysfunction of motor, sensory, and visual systems with only rare involvement of higher cortical function. Among symptoms reported, visual abnormalities are the most variable and the least common. However, the visual field abnormality may be overlooked and the incidence underestimated since some patients may not be aware of the problem until uncovered by formal visual field testing. PMID- 16697645 TI - Disseminated spread of recurrent oligodendroglioma (WHO grade II). AB - Oligodendroglioma is a relatively uncommon primary brain tumour. The occurrence of metastatic dissemination of oligodendroglioma is rare and usually occurs in patients with anaplastic oligodendroglioma. The dissemination of WHO Grade II oligodendroglioma can occur and we report a patient with an initial diagnosis of a left temporal oligodendroglioma who presented with disseminated disease in the left temporal lobe, sellar region, medulla oblongata, both frontal lobes and ventricles more than 8 years later. Histology at dissemination showed anaplastic oligodendroglioma. Similar reports of metastatic dissemination of oligodendrogliomas reveal that the tumours may remain as WHO Grade II or may progress to anaplastic oligodendroglioma at metastasis. However, regardless of the histological grade at metastasis, the prognosis of metastatic oligodendroglioma is poor. PMID- 16697646 TI - Discovery of new chemical leads for selective EP1 receptor antagonists. AB - A series of 4-([2-[alkyl(phenylsulfonyl)amino]phenoxy]methyl)benzoic acids were identified as functional PGE(2) antagonists with selectivity for the EP1 receptor subtype starting from a chemical lead 1, which was found while screening our in house compound library. Discovery of the optimized analogs 21-23 is presented here and structure-activity relationships (SAR) are also discussed. PMID- 16697647 TI - Structure-activity studies on the protection of Trimetazidine derivatives modified with nitroxides and their precursors from myocardial ischemia reperfusion injury. AB - Trimetazidine, the known anti-anginal and anti-ischemic drug, was modified by pyrroline and tetrahydropyridine nitroxides and their hydroxylamine and sterically hindered secondary amine precursors. The synthesized new compounds proved to be better superoxide scavenger molecules compared to the parent Trimetazidine in an in vitro experiment. This reactive oxygen species (ROS) scavenging activity was further supported by ischemia/reperfusion (I/R) studies on Langendorff-perfused rat hearts pretreated with Trimetazidine and with the modified Trimetazidine derivatives before ischemia. Two of the investigated compounds, containing 2,2,5,5-tetramethyl-2,5-dihydro-1H-pyrrole and 4-phenyl 2,2,5,5-tetramethyl-2,5-dihydro-1H-pyrrole substituents on the piperazine ring, provided significant protection from the cardiac dysfunction caused by I/R. The protective effect could be attributed to the combined anti-ischemic and antioxidant effects. PMID- 16697648 TI - Adsorption behavior of strontium on binary mineral mixtures of montmorillonite and kaolinite. AB - The adsorption behavior of kaolinite and montmorillonite minerals and their mixtures in respect of Sr ion were studied by means of a batch method using (90)Sr as a radio tracer. The effect of several parameters such as temperature, pH, Sr concentration and supporting electrolyte were investigated. Experimentally measured distribution coefficients showed a good agreement to within 98.5-99.7% with theoretically calculated values. The values of adsorption capacity of adsorbents and mean adsorption energy of adsorption were calculated by fitting the adsorption data to Dubinin-Radushkevich isotherm. The adsorption capacity of clay mixtures decreased as kaolinite fractions increased. The mean adsorption energy values of 8.0-9.5 kJ mol(-1) showed that adsorption was governed by ion exchange. The Freundlich parameters were used to characterize a site distribution function for binary exchange between Sr and Na. PMID- 16697649 TI - Application of high-resolution radiochromic film dosimetry in verifying a small field stereotactic radiosurgery plan. AB - A high-resolution radiochromic film dosimetry (Hr-RCFD) method has been applied to verify a small-field stereotactic radiosurgery (SRS) plan. This was done by exposing a RCF in a Perspex head phantom undergoing the same treatment plan as the patient. The dose distribution obtained by the Hr-RCFD was verified against that calculated by the stereotactic treatment planning system and the result was satisfactory. The Hr-RCFD method has been found to be an accurate and practical tool in verifying small-field SRS plans. PMID- 16697650 TI - Increased vulnerability of ApoE4 neurons to HIV proteins and opiates: protection by diosgenin and L-deprenyl. AB - Human immunodeficiency virus (HIV) infection continues to rise in drug-abusing populations and causes a dementing illness in a subset of individuals. Factors contributing to the development of dementia in this population remain unknown. We found that HIV-infected individuals with the E4 allele of Apolipoprotein E (ApoE) or history of intravenous drug abuse had increased oxidative stress in the CNS. In vitro studies showed that HIV proteins, gp120 and Tat, Tat + morphine but not tumor necrosis factor-alpha (TNF-alpha), caused increased neurotoxicity in human neuronal cultures with ApoE4 allele. Microarray analysis showed a differential alteration of transcripts involved in energy metabolism in cultures of ApoE3 and 4 neurons upon treatment with Tat + morphine. This was confirmed using assays of mitochondrial function and exposure of the neurons to Tat + morphine. Using this in vitro model, we screened a number of novel antioxidants and found that only L deprenyl and diosgenin protected against the neurotoxicity of Tat + morphine. Furthermore, Tat-induced oxidative stress impaired morphine metabolism which could also be prevented by diosgenin. In conclusion, opiate abusers with HIV infection and the ApoE4 allele may be at increased risk of developing dementia. L deprenyl and a plant estrogen, diosgenin, may have therapeutic potential in this population. PMID- 16697651 TI - Adriamycin-induced, TNF-alpha-mediated central nervous system toxicity. AB - The clinical effectiveness of adriamycin (ADR), a potent chemotherapeutic, is known to be limited by severe cardiotoxic side effects. However, the effect of ADR on brain tissue is not well understood. It is generally thought that ADR is not toxic to the brain because ADR does not pass the blood-brain barrier. The present study demonstrates that ADR autofluorescence was detected only in areas of the brain located outside the blood-brain barrier, but a strong tumor necrosis factor (TNF) alpha immunoreactivity was detected in the cortex and hippocampus of ADR-treated mice. Systemic injection of ADR led to a decline in brain mitochondrial respiration via complex I substrate shortly after ADR treatment (P < 0.05). Cytochrome c release, increased caspase 3 activity, and TUNEL-positive cell death all were suggestive of apoptosis in brain following systemic ADR treatment. The levels of the known pro-apoptotic proteins, p53 and Bax, were increased in brain mitochondria at 3 h following ADR treatment and declined by 48 h. In contrast, the anti-apoptotic protein, Bcl-xL, was increased later at 6 h post-ADR treatment and was sustained throughout 72 h. Furthermore, p53 migrated to mitochondria and interacted with Bcl-xL, supporting the hypothesis that mitochondria are targets of ADR-induced CNS injury. Neutralizing antibodies against circulating TNF completely abolished both the increased TNF in the brain and the observed mitochondrial injury in brain tissues. These results are consistent with the notion that TNF is an important mediator by which ADR induces central nervous system (CNS) injury. This study, the first to provide direct biochemical evidence of ADR toxicity to the brain, revealed novel mechanisms of ADR-induced CNS injury and suggests a potential therapeutic intervention against circulating TNF-induced CNS effects. PMID- 16697652 TI - Elevated brain 3-hydroxykynurenine and quinolinate levels in Huntington disease mice. AB - The brain levels of the endogenous excitotoxin quinolinic acid (QUIN) and its bioprecursor, the free radical generator 3-hydroxykynurenine (3-HK), are elevated in early stage Huntington disease (HD). We now examined the status of these metabolites in three mouse models of HD. In R6/2 mice, 3-HK levels were significantly and selectively elevated in the striatum, cortex and cerebellum starting at 4 weeks of age. In contrast, both 3-HK and QUIN levels were increased in the striatum and cortex of the full-length HD models, beginning at 8 months (YAC128) and 15 months (Hdh(Q92) and Hdh(Q111)), respectively. No changes were seen in 13-month-old shortstop mice, which show no signs of motor or cognitive dysfunction or selective neuropathology. These results demonstrate both important parallels and intriguing differences in the progressive neurochemical changes in these HD mouse models and support the hypothesis that QUIN may play a role in the striatal and cortical neurodegeneration of HD. PMID- 16697653 TI - Biotransformation of mycosporine like amino acids (MAAs) in the toxic dinoflagellate Alexandrium tamarense. AB - Changes in mycosporine-like amino acids (MAAs) induced by the increase of photosynthetically active radiation (PAR) were studied in the toxic dinoflagellate Alexandrium tamarense. Cultures of A. tamarense were maintained at exponential growth under low (25 micromol quanta m(-2)s(-1)) PAR irradiance. The cultures were nutrient enriched and one day later exposed to higher irradiance (150 micromol quanta m(-2)s(-1)). The content of MAAs was determined by means of high performance liquid chromatography (HPLC). Eleven MAAs, including some partially characterized compounds, were identified. The MAAs synthesis induction can be described as a two-stage process. The first one involves the net synthesis of the MAAs bi-substituted by amino acids. In the second stage these compounds were transformed into other secondary MAAs. The two most prominent changes were observed in the concentration of porphyra-334 and palythene. The cellular concentration of porphyra-334 increased during the first 2h of exposure to higher irradiance and then decreased rapidly. In contrast, the cellular concentration of palythene showed a continuous accumulation since the beginning of the exposure. In A. tamarense the main route of MAAs transformation has porphyra-334 as a precursor of a sequential conversion resulting in the accumulation of palythene. PMID- 16697654 TI - HIV-infected patients with lipodystrophy have higher rates of carotid atherosclerosis: the role of monocyte chemoattractant protein-1. AB - Individuals with HIV-1 infection are at increased risk for cardiovascular events, and lipodystrophy is generally associated with pro-atherogenic metabolic disturbances. We conducted a case-control study to assess the presence of sub clinical atherosclerosis in HIV-1-infected patients with or without lipodystrophy (LD) and to evaluate the influence of monocyte chemoattractant protein-1 (MCP-1) on the development of both carotid atherosclerosis and LD. The study population consisted of 43 patients with LD and 86 patients without LD. We determined carotid intima-media thickness (IMT), MCP-1 concentrations in plasma, and MCP-1 genotype (presence or absence of the -2518G allele). HIV-1-infected patients with LD showed increased risk (OR=3.71, 95% CI=1.10-12.47, p=0.03) for sub-clinical atherosclerosis, and MCP-1 plasma concentration was significantly correlated with IMT in these patients (Pearson=0.31, p=0.03). Furthermore, presence of LD was a determinant for MCP-1 plasma concentration (beta=0.18, p=0.05). In summary, HIV-1 infected patients with clinically manifest LD are at higher risk for atherosclerosis and our observations support the relationship between inflammation and atherosclerotic disease. PMID- 16697655 TI - Study of cytokines involved in the prevention of a murine experimental breast cancer by kefir. AB - Previous studies have shown that compounds released during milk fermentation by Lactobacillus helveticus are implicated in the antitumour effect of this product. Here the effects of the consumption, during 2 or 7 days, of kefir or kefir cell free fraction (KF) on the systemic and local immune responses in mammary glands and tumours using a murine hormone-dependent breast cancer model were studied. In the tumour control group, mice did not receive these products. At the end of the feeding period, mice were injected subcutaneously with tumour cells in the mammary gland. Four days post-injection, they received kefir or KF on a cyclical basis. Rate of tumour development, cytokines in serum; mammary gland tissue, and tumour isolated cells were monitored. Two-day cyclical administration of both products delayed tumour growth. Both kefir and KF increased IL-10 in serum and decreased IL-6(+) cells (cytokine involved in oestrogen synthesis) in mammary glands. Two-day cyclical administration of KF increased IL-10(+) cells in mammary glands and in tumours and decreased IL-6(+) cells in tumour. This study demonstrated the modulatory capacity of KF on the immune response in mammary glands and tumours and the importance of the administration period to obtain this effect. PMID- 16697656 TI - Integrin alpha 11 is a novel type I interferon stimulated gene. AB - Interferons (IFNs) are a family of cytokines that have many biological functions in the cell, including regulation of cellular growth, differentiation, immunomodulation, and viral replication by inducing a set of interferon stimulated genes (ISGs). Based on their structure and biological activities IFNs are subdivided into two groups: type I IFNs, which includes IFN-alpha and IFN beta and type II IFNs, represented by IFN-gamma. The aim of this work was to investigate whether integrin alpha 11 (ITGA-11), a novel collagen-binding integrin, is responsive to type I IFN treatment. Our findings indicated that type I IFNs were able to induce the ITGA-11 mRNA levels in T98G cells. Increased levels of ITGA-11 protein were also observed in IFN-treated cells. The in vivo induction of ITGA-11 was detected in spleen and lungs of IFN-treated BALB/c mice. T98G cells infected with Murine encephalomyocarditis virus showed increased levels of ITGA-11 mRNA and protein. We observed that the ITGA-11 promoter has binding sites for transcriptional factors regulated by IFNs and the double stranded RNA dependent protein kinase (PKR). Therefore we investigated the role of PKR in the induction of ITGA-11 by using a PKR deficient mouse embryo fibroblast cell line (MEFs). PKR(-/-) MEFs treated with IFN did not show increased levels of ITGA-11 protein nor mRNA although that could be promptly detected in wild type MEFs. Taken together our data suggest that ITGA-11 is a new interferon stimulated gene. PMID- 16697657 TI - Effect of overuse of the antimigraine combination of indomethacin, prochlorperazine and caffeine (IPC) on the disposition of its components in chronic headache patients. AB - BACKGROUND: The combination of indomethacin, prochlorperazine and caffeine (IPC) is one of the most utilized formulations for the treatment of migraine attacks in Italy. Several patients suffering from chronic headache overuse this symptomatic medication in the attempt to control their headache. OBJECTIVE: To verify whether overuse of IPC combination by chronic headache patients is associated with modified disposition of its components. METHODS: We studied indomethacin, prochlorperazine, and caffeine disposition in 34 female subjects suffering from primary headaches, subdivided into four groups: eight migraine patients occasionally using IPC combination suppositories-group 1; nine patients with chronic headache and probable medication-overuse headache, daily taking one or more suppositories of the IPC combination-group 2; 11 migraine patients occasionally using "mild" suppositories of the IPC combination-group 3; six migraine patients occasionally taking tablets of the IPC combination-group 4. The IPC combination habitually used was administered to each patient. Blood samples were taken at baseline and at fixed intervals up to 6h after administration. Plasma levels of indomethacin and prochlorperazine were assayed by high-pressure liquid chromatographic (HPLC) method; caffeine levels were assayed by enzyme multiplied immunoassay test (EMIT). Pharmacokinetic parameters were calculated by means of a computer software (P K Solutions 2.0. Summit Research Services, Montrose, CO, USA). RESULTS: Half-life of indomethacin was longer, and clearance lower, in group 2 than in the other groups; AUC of indomethacin in group 2 was twice that in group 1 (P<0.05, Newman-Keuls' test). Peak concentrations and AUC(0 ->infinity) of caffeine were significantly higher in group 2 than in the other groups (P<0.05, Newman-Keuls' test). We could not define prochlorperazine disposition because it was not detectable in the majority of blood samples. CONCLUSION: Overuse of IPC combination in chronic headache patients is associated with increased plasma levels of indomethacin and caffeine, and with delayed elimination of indomethacin; the high and sustained concentrations of these drugs may cause rebound headache, organ damages, and perpetuate medication-overuse headache. PMID- 16697658 TI - Determination of cooling rates in a quadrupole ion trap. AB - Collisional cooling rates of infrared excited ions are measured in a quadrupole ion trap (QIT) mass spectrometer at different combinations of temperature and pressure. Measurements are carried out by monitoring fragmentation efficiency of leucine enkephalin as a function of irradiation time by an infrared laser after a short excitation and incrementally increasing cooling periods. Cooling rates are observed to be directly related to bath gas pressure and inversely related to bath gas temperature. The cooling rate at typical ion trap operating pressure (1 mTorr) and temperature (room T) is faster than can be measured. At elevated temperature and the lowest pressure used for the studies, the rate of collisional cooling becomes negligible compared to the rate of radiative cooling. PMID- 16697659 TI - Miniature toroidal radio frequency ion trap mass analyzer. AB - A miniature ion trap mass analyzer is reported. The described analyzer is a 1/5 scale version of a previously reported toroidal radio frequency (rf) ion trap mass analyzer. The toroidal ion trap operates with maximum rf trapping voltages about 1 kVp-p or less; however despite the reduced dimensions, it retains roughly the same ion trapping capacity as conventional 3D quadrupole ion traps. The curved geometry provides for a compact mass analyzer. Unit-mass resolved mass spectra for n-butylbenzene, xenon, and naphthalene are reported and preliminary sensitivity data are shown for naphthalene. The expected linear mass scale with rf amplitude scan is obtained when scanned using a conventional mass-selective instability scan mode combined with resonance ejection. PMID- 16697660 TI - Expanded use of a battery-powered two-electrode emitter cell for electrospray mass spectrometry. AB - A battery-powered, controlled-current, two-electrode electrochemical cell containing a porous flow-through working electrode with high surface area and multiple auxiliary electrodes with small total surface area was incorporated into the electrospray emitter circuit to control the electrochemical reactions of analytes in the electrospray emitter. This cell system provided the ability to control the extent of analyte oxidation in positive ion mode in the electrospray emitter by simply setting the magnitude and polarity of the current at the working electrode. In addition, this cell provided the ability to effectively reduce analytes in positive ion mode and oxidize analytes in negative ion mode. The small size, economics, and ease of use of such a battery-powered controlled current emitter cell was demonstrated by powering a single resistor and switch circuit with a small-size, 3 V watch battery, all of which might be incorporated on the emitter cell. PMID- 16697661 TI - Role of PTPN22 in type 1 diabetes and other autoimmune diseases. AB - We recently discovered that a single-nucleotide polymorphism (SNP) in the lymphoid tyrosine phosphatase (LYP), encoded by the PTPN22 gene on chromosome 1p13, correlates strongly with the incidence of type 1 diabetes (T1D) in two independent populations. This findings has now been verified by numerous studies and it has been expanded to rheumatoid arthritis, juvenile rheumatoid arthritis, systemic lupus erythematosus, Graves' disease, generalized vitiligo and other autoimmune disease. Here we review the genetics of the SNP and its association with autoimmunity, discuss the function of the phosphatase in signaling, the biochemistry of the disease-predisposing allele, and the possible mechanisms by which PTPN22 contributes to the development of human disease. PMID- 16697662 TI - 14-3-3 proteins in cell cycle regulation. AB - Co-ordinated progression through the cell cycle is essential for the maintenance of genomic integrity. Several checkpoint mechanisms guarantee that the next step in cell cycle progression is only entered after error-free completion of the previous phase. Cell cycle deregulation caused by changes in 14-3-3 expression has been implicated in cancer formation. 14-3-3 proteins function at several key points in G(1)/S- and G(2)/M-transition by binding to regulatory proteins and modulating their function. In most cases, the association with 14-3-3 proteins requires a specific phosphorylation of the protein ligand and mediates cell cycle arrest. 14-3-3 binding may lead to cytoplasmic sequestration of the protein ligand but may also have other functional consequences. The 14-3-3sigma gene is induced by p53 and its product inhibits G(2)/M progression by cytoplasmatic sequestration of CDC2-cyclin B complexes. In addition, 14-3-3 proteins have been implicated in the transcriptional regulation of CDK-inhibitors as they modulate the transcription factors p53, FOXO and MIZ1. Effects of 14-3-3 proteins on cell cycle progression and the regulation of 14-3-3 activity during the cell cycle are reviewed in this chapter. PMID- 16697663 TI - Reelin is transiently expressed in the peripheral nerve during development and is upregulated following nerve crush. AB - Reelin is an extracellular matrix protein which is critical for the positioning of migrating post-mitotic neurons and the laminar organization of several brain structures during development. We investigated the expression and localization of Reelin in the rodent peripheral nerve during postnatal development and following crush injury in the adult stage. As shown with Western blotting, immunocytochemistry and RT-PCR, Schwann cells in the developing peripheral nerve and in primary cultures from neonatal nerves produce and secrete Reelin. While Reelin levels are downregulated in adult stages, they are again induced following sciatic nerve injury. A morphometric analysis of sciatic nerve sections of reeler mice suggests that Reelin is not essential for axonal ensheathment by Schwann cells, however, it influences the caliber of myelinated axons and the absolute number of fibers per unit area. This indicates that Reelin may play a role in peripheral nervous system development and repair by regulating Schwann cell-axon interactions. PMID- 16697664 TI - The neural basis of the hemodynamic response nonlinearity in human primary visual cortex: Implications for neurovascular coupling mechanism. AB - It has been well recognized that the nonlinear hemodynamic responses of the blood oxygenation level-dependent (BOLD) functional MRI (fMRI) are important and ubiquitous in a series of experimental paradigms, especially for the event related fMRI. Although this phenomenon has been intensively studied and it has been found that the post-capillary venous expansion is an intrinsically nonlinear mechanical process, the existence of an additional neural basis for the nonlinearity has not been clearly shown. In this paper, we assessed the correlation between the electric and vascular indices by performing simultaneous electroencephalography (EEG) and fMRI recordings in humans during a series of visual stimulation (i.e., radial checkerboard). With changes of the visual stimulation frequencies (from 0.5 to 16 Hz) and contrasts (from 1% to 100%), both the event related potentials (ERPs) and hemodynamic responses show nonlinear behaviors. In particular, the mean power of the brain electric sources and the neuronal efficacies (as originally defined in the hemodynamics model [Friston et al. Neuroimage, 12, 466-477, 2000], here represent the vascular inputs) in primary visual cortex consistently show a linear correlation for all subjects. This indicates that the hemodynamic response nonlinearity found in this paper primarily reflects the nonlinearity of underlying neural activity. Most importantly, this finding underpins a nonlinear neurovascular coupling. Specifically, it is shown that the transferring function of the neurovascular coupling is likely a power transducer, which integrates the fast dynamics of neural activity into the vascular input of slow hemodynamics. PMID- 16697665 TI - Hierarchical microimaging for multiscale analysis of large vascular networks. AB - There is a wide range of diseases and normal physiological processes that are associated with alterations of the vascular system in organs. Ex vivo imaging of large vascular networks became feasible with recent developments in microcomputed tomography (microCT). Current methods permit to visualize only limited numbers of physically excised regions of interests (ROIs) from larger samples. We developed a method based on modified vascular corrosion casting (VCC), scanning electron microscopy (SEM), and desktop and synchrotron radiation microCT (SRmicroCT) technologies to image vasculature at increasing levels of resolution, also referred to as hierarchical imaging. This novel approach allows nondestructive 3D visualization and quantification of large microvascular networks, while retaining a precise anatomical context for ROIs scanned at very high resolution. Scans of entire mouse brain VCCs were performed at 16-microm resolution with a desktop microCT system. Custom-made navigation software with a ROI selection tool enabled the identification of anatomical brain structures and precise placement of multiple ROIs. These were then scanned at 1.4-microm voxel size using SRmicroCT and a local tomography setup. A framework was developed for fast sample positioning, precise selection of ROIs, and sequential high-throughput scanning of a large numbers of brain VCCs. Despite the use of local tomography, exceptional image quality was achieved with SRmicroCT. This method enables qualitative and quantitative assessment of vasculature at unprecedented resolution and volume with relatively high throughput, opening new possibilities to study vessel architecture and vascular alterations in models of disease. PMID- 16697666 TI - Skull-stripping magnetic resonance brain images using a model-based level set. AB - The segmentation of brain tissue from nonbrain tissue in magnetic resonance (MR) images, commonly referred to as skull stripping, is an important image processing step in many neuroimage studies. A new mathematical algorithm, a model-based level set (MLS), was developed for controlling the evolution of the zero level curve that is implicitly embedded in the level set function. The evolution of the curve was controlled using two terms in the level set equation, whose values represented the forces that determined the speed of the evolving curve. The first force was derived from the mean curvature of the curve, and the second was designed to model the intensity characteristics of the cortex in MR images. The combination of these forces in a level set framework pushed or pulled the curve toward the brain surface. Quantitative evaluation of the MLS algorithm was performed by comparing the results of the MLS algorithm to those obtained using expert segmentation in 29 sets of pediatric brain MR images and 20 sets of young adult MR images. Another 48 sets of elderly adult MR images were used for qualitatively evaluating the algorithm. The MLS algorithm was also compared to two existing methods, the brain extraction tool (BET) and the brain surface extractor (BSE), using the data from the Internet brain segmentation repository (IBSR). The MLS algorithm provides robust skull-stripping results, making it a promising tool for use in large, multi-institutional, population-based neuroimaging studies. PMID- 16697667 TI - Isolation of erythroid cells from the mouse embryonic yolk sac by laser capture microdissection and subsequent microarray hybridization. AB - Erythropoietic tissues are complex, containing both erythroid and other cells. The embryonic yolk sac in particular contains primitive erythroid cells in low abundance. Laser capture microdissection (LCM) was performed to isolate erythroid cells, and epithelial cells, from mouse embryonic day 10 (E10) yolk sac. Quantitative RT-PCR was performed to confirm that enriched cell populations were obtained. epsilony- and betaH1-globin mRNAs were enriched in the erythroid compared to the epithelial fraction, and villin mRNA was enriched in the epithelial compared to the erythroid fraction. RNA isolated from the microdissected erythroid cells was of high quality as indicated by capillary electrophoresis. The RNA from the LCM erythroid fraction was linearly amplified with T7 RNA polymerase and hybridized to a Mouse 430A 2.0 Affymetrix array. Forty eight percent of genes were present in the microarray assays, including low abundance transcripts such as erythroid transcription factors and enzymes involved in heme synthesis. With the LCM/microarray strategy, it will be possible to identify genes that are differentially regulated in native primitive and definitive erythroid cells. PMID- 16697668 TI - Membrane mobility and clustering of Integrin Associated Protein (IAP, CD47)- major differences between mouse and man and implications for signaling. AB - Integrin Associated Protein (IAP, CD47) is a ubiquitous integral membrane protein implicated in processes (in mice) that range from inhibiting clearance by phagocytes [Oldenborg et al., Science 2000; Gardai et al., Cell 2005] to neutrophil motility [Lindberg et al., Science 1996]. SIRPalpha is CD47's main receptor on phagocytes plus a number of other cell types, and SIRPalpha-CD47 interactions in clusters are believed to mediate signaling. However, considerable species differences in CD47 sequence as well as differences in CD47 extractability from mouse cells versus man motivate a characterization of mobility, clusterability, and kinetics under force of CD47-SIRPalpha. Despite similar levels of CD47 on red cells from mouse and man, we find an effective avidity of SIRPalpha-CD47 for mouse appears higher than for human. Both mouse and human CD47 show clustering by multivalent SIRPalpha complexes, but only mouse cells aggregate with CD47 concentrating at cell-cell contacts. This proves consistent with fluorescence imaged micro-deformation, which indicates near complete mobility of CD47 on mouse cells compared to only about 30-40% mobility on normal human cells. To qualify the method, we also show that disrupting cellular F-actin dramatically increases the mobility of integral membrane proteins. Furthermore, atomic force microscopy probing of cell membranes with human SIRPalpha confirms the species-specific interactions and provides evidence of clustering and adhesion on short time scales, but it also shows surprisingly strong forces in detachment for a signaling complex. The results thus highlight major species differences in CD47-SIRPalpha interactions and CD47 integration, suggesting that signaling by CD47 in man may be qualitatively different from mouse. PMID- 16697669 TI - Phase-specific gene expression underlying morphological adaptations of the dimorphic human pathogenic fungus, Coccidioides posadasii. AB - Coccidioides posadasii is a dimorphic fungal pathogen that grows as a filamentous saprobe in the soil and as endosporulating spherules within the host. To identify genes specific to the pathogenic phase of Co. posadasii, we carried out a large scale study of gene expression in two isolates of the species. From the sequenced Co. posadasii genome, we chose 1,000 open reading frames to construct a 70-mer microarray. RNA was recovered from both isolates at three life-cycle phases: hyphae, presegmented spherules, and spherules releasing endospores. Comparative hybridizations were conducted in a circuit design, permitting comparison between both isolates at all three life-cycle phases, and among all life-cycle phases for each isolate. By using this approach, we identified 92 genes that were differentially expressed between pathogenic and saprobic phases in both fungal isolates, and 43 genes with consistent differential expression between the two parasitic developmental phases. Genes with elevated expression in the pathogenic phases of both isolates included a number of genes that were involved in the response to environmental stress as well as in the metabolism of lipids. The latter observation is in agreement with previous studies demonstrating that spherules contain a higher proportion of lipids than saprobic phase tissue. Intriguingly, we discovered statistically significant and divergent levels of gene expression between the two isolates profiled for 64 genes. The results suggest that incorporating more than one isolate in the experimental design offers a means of categorizing the large collection of candidate genes that transcriptional profiling typically identifies into those that are strain specific and those that characterize the entire species. PMID- 16697670 TI - A jack-in-the-box of respiratory research: is the technique of barometric whole body plethysmography a disappointing surprise? PMID- 16697671 TI - Efficient orientational averaging by the extension of Lebedev grids via regularized octahedral symmetry expansion. AB - Gaussian spherical quadrature methods in the guise of the Lebedev sampling grids are highly efficient for some orientational ("powder") averaging problems in solid state NMR. However, their applicability is currently restricted, as the sets of orientations are derived analytically and because they are not well adapted to simulate the broad peakshapes encountered, for example, in the NMR on static powders or on half-integer quadrupolar spins subject to second order quadrupolar interactions under magic-angle spinning conditions. We remedy these problems by (i) introducing the recursive procedure regularized octahedral symmetry expansion (ROSE), to which any existing Lebedev set may be subjected. Each recursive step gives a 9-fold enlarged set of orientations. (ii) We demonstrate that ROSE-expanded grids, in conjunction with spectral interpolation, is well suited for calculating broad peakshapes. These advances combine into the apparently most efficient general-purpose two-angle orientational averaging technique proposed to date for solid state NMR applications. PMID- 16697672 TI - Comparative neuroendocrinology: Integration of hormonal and environmental signals in vertebrates and invertebrates. PMID- 16697673 TI - Hypopituitarism after acute brain injury. AB - Acute brain injury has many causes, but the most common is trauma. There are 1.5 2.0 million traumatic brain injuries (TBI) in the United States yearly, with an associated cost exceeding 10 billion dollars. TBI is the most common cause of death and disability in young adults less than 35 years of age. The consequences of TBI can be severe, including disability in motor function, speech, cognition, and psychosocial and emotional skills. Recently, clinical studies have documented the occurrence of pituitary dysfunction after TBI and another cause of acute brain injury, subarachnoid hemorrhage (SAH). These studies have consistently demonstrated a 30-40% occurrence of pituitary dysfunction involving at least one anterior pituitary hormone following a moderate to severe TBI or SAH. Growth hormone (GH) deficiency is the most common pituitary hormone disorder, occurring in approximately 20% of patients when multiple tests of GH deficiency are used. Within 7-21 days of acute brain injury, adrenal insufficiency is the primary concern. Pituitary function can fluctuate over the first year after TBI, but it is well established by 1 year. Studies are ongoing to assess the effects of hormone replacement on motor function and cognition in TBI patients. Any subject with a moderate to severe acute brain injury should be screened for pituitary dysfunction. PMID- 16697674 TI - Innate immunity in plants: a continuum of layered defenses. AB - Plant responses to pathogenic invaders result from recognition of nonself elicitors. Host surveillance proteins activate distinct signaling pathways that induce partially overlapping defensive responses. Pathogen virulence is promoted by inhibition of these pathways. This evolutionary struggle has produced plant immune systems that rely on a continuum of layered defenses. PMID- 16697675 TI - HIV tat and neurotoxicity. AB - HIV tat is the transactivator of HIV-1, supporting efficient viral replication by stabilizing the transcription of viral genes. Tat can be released from HIV infected cells and alter several functions in uninfected cells. In the brain, tat induces neuronal dysfunction/toxicity, even though neurons cannot be directly infected with HIV, resulting in CNS pathology, such as the dementia and encephalitis associated with NeuroAIDS. This review discusses the most recent data addressing tat-induced neurotoxicity and integrates these new findings in the context of NeuroAIDS. PMID- 16697676 TI - Dual function of the long pentraxin PTX3 in resistance against pulmonary infection with Klebsiella pneumoniae in transgenic mice. AB - The long pentraxin PTX3 is expressed during acute inflammation and appears to control nitric oxide (NO) and tumor necrosis factor (TNF)-alpha production. In the present study, the physiological function of PTX3 was investigated in a model of pulmonary infection caused by the Gram-negative bacterium Klebsiella pneumoniae. Transgenic mice expressing multiple copies of PTX3 under the control of its own promoter were used to assess lethality rates, bacterial counts and inflammatory indices following pulmonary infection of mice. Expression of PTX3 is enhanced during pulmonary infection in wild-type mice. In transgenic mice given a high inoculum, overt PTX3 expression was associated with faster lethality. Faster lethality correlated with enhanced nitrate in plasma, an inability of neutrophils to migrate to lung tissue and greater dissemination of bacteria to blood at 20h after infection. In contrast, transgenic PTX3 expression conferred protection to mice given lower pulmonary inocula. In the latter experiments, there was enhanced TNF-alpha production, greater neutrophil influx and phagocytosis of bacteria by migrated neutrophils. By controlling the production of TNF-alpha and NO, and depending on the intensity of the inflammatory response induced by a given inoculum, the expression of PTX3 may favor or disfavor the influx of neutrophils and the ability of the murine host to deal with pulmonary infection with K. pneumoniae. These experiments highlight the delicate balance that exists among the various mediators that control the inflammatory response and suggest that PTX3 is an essential part of the ability of a host to deal with bacterial infection. PMID- 16697677 TI - Current understanding of the genetic diversity of Mycobacterium avium subsp. paratuberculosis. AB - Mycobacterium avium subsp. paratuberculosis (MAP) is the etiological agent of Johne's disease (or paratuberculosis). Paratuberculosis is a chronic gastroenteritis mainly affecting cattle, sheep and other ruminants. MAP is also of concern due to the heretofore unresolved issue of its possible role in Crohn's disease in humans. We present here a review of MAP (i) mobile genetic elements; (ii) repetitive elements; (iii) single nucleotide polymorphisms; and (iv) whole genome comparisons to study the molecular epidemiology of MAP. A summary of the findings to date is presented, and the discriminatory power, advantage and disadvantages of each of the methods are compared and discussed. PMID- 16697678 TI - Induction of beta-defensin 3 in keratinocytes stimulated by bacterial lipopeptides through toll-like receptor 2. AB - The epidermis, which covers the surface of all mammals, serves as a front line of defense against the invasion of pathogenic microbes and acts as a crucial site for innate immune responses. Various antimicrobial molecules are expressed not only on the surfaces of monocytes but also on epithelial cells. beta-Defensins, a family of antimicrobial peptides, are produced by several types of epithelial cells, including keratinocytes. However, the induction pathways for beta defensins in keratinocytes are not fully understood. We hypothesized that bacterial components would trigger the expression of beta-defensins in keratinocytes through a toll-like receptor (TLR)-MyD88 signaling pathway that plays important roles in innate immunity. Production of TNF-alpha and IL-1 alpha following stimulation with lipopolysaccharide or bacterial lipopeptides was completely abolished in TLR2&TLR4-doubly deficient keratinocytes and in MyD88 deficient keratinocytes. Expression of murine beta-defensin was upregulated by bacterial lipopeptides in wild-type keratinocytes, while it was attenuated in TLR2-deficient keratinocytes. To evaluate the in vivo role of TLRs in keratinocytes, we inoculated Staphylococcus aureus into the tail skin from TLR2 deficient mice that had been grafted on the dorsal skin of syngeneic mice. The grafted skin from TLR2-deficient mice resulted in erosion. These studies strongly suggest that the TLR2-MyD88-dependent pathway in keratinocytes is essential for antimicrobial activity in vivo. PMID- 16697679 TI - Development of a novel Borna disease virus reverse genetics system using RNA polymerase II promoter and SV40 nuclear import signal. AB - Borna disease virus (BDV) is a noncytolytic, neurotropic RNA virus that replicates and transcribes in the nucleus of infected cells. Therefore, efficient synthesis of BDV RNA in the nucleus is critical for the development of a reverse genetics system for this virus. Here, we report the development of such a system using the RNA polymerase II (Pol II) promoter. The BDV minigenome cDNA was flanked by hammerhead ribozyme and hepatitis delta ribozyme sequences and inserted downstream of the Pol II promoter. To improve the efficacy of minigenome expression, we estimated the effects of several signal sequences within the minigenome constructs. We found that insertion of the SV40 nuclear import sequence into the Pol II constructs significantly enhances the replication of the minigenome even in cells lacking the SV40 large T antigen. This novel system is theoretically applicable to any mammalian cell line and would be valuable for analyzing host- or cell-type-dependent differences in BDV replication and production. We could demonstrate here the cell-type-dependent inhibitory effect of the viral protein X on BDV polymerase activity. This system may be useful for various research fields not only of BDV but also of other negative-sense RNA viruses. PMID- 16697680 TI - Effect of porcine respiratory coronavirus infection on lipopolysaccharide recognition proteins and haptoglobin levels in the lungs. AB - Porcine respiratory coronavirus (PRCV) potentiates respiratory disease and proinflammatory cytokine production in the lungs upon intratracheal inoculation with lipopolysaccharide (LPS) at 1 day of infection. This study aimed to quantify LPS-binding protein (LBP), CD14 and haptoglobin in the lungs throughout a PRCV infection. LBP and CD14 recognize LPS and enhance its endotoxic activity, whereas haptoglobin dampens it. Gnotobiotic pigs were inoculated intratracheally with PRCV (n = 34) or saline (n = 5) and euthanized 1-15days post inoculation (DPI). Virus was detected in the lungs from 1 to 9DPI. Cell-associated CD14 in lung tissue increased up to 15 times throughout the infection, due to an increase in highly CD14+ monocyte-macrophages from 1 to 12DPI and CD14+ type 2 pneumocytes from 7 to 9DPI. LBP and soluble CD14 levels in bronchoalveolar lavage fluids were elevated from 1-12DPI, with up to 35- and 4-fold increases, respectively. Haptoglobin levels increased significantly (x4.5) at 7DPI. In addition, we found that PRCV could sensitize the lungs to LPS throughout the infection, but the response to LPS appeared less enhanced at the end of infection (7DPI). The marked increases in LBP, CD14 and haptoglobin were not correlated with the extent of the LPS response. PMID- 16697681 TI - Host-cell interaction of attenuated and wild-type strains of yellow fever virus can be differentiated at early stages of hepatocyte infection. AB - Yellow fever (YF) virus is currently found in tropical Africa and South America, and is responsible for a febrile to severe illness characterized by organ failure and shock. The attenuated YF 17D strain, used in YF vaccine, was derived from the wild-type strain Asibi. Although studies have been done on genetic markers of YF virulence, differentiation of the two strains in terms of host-cell interaction during infection remains elusive. As YF wild-type strains are hepatotropic, we chose a hepatic cell line (HepG2) to study YF virus-host cell interaction. HepG2 cells rapidly produced high titres of infectious viral particles for 17D and Asibi YF strains. However, HepG2 cells were more susceptible to the attenuated 17D virus infection, and only this virus strain induced early apoptosis in these cells. Molecular markers specific for the 17D virus were identified by microarray analysis and confirmed by quantitative RT-PCR analysis. As early as 1h postinfection, three genes, (IEX-1, IRF-1, DEC-1) all implicated in apoptosis pathways, were upregulated. Later in infection (48 h) two other genes (HSP70-1A and 1B), expressed in cases of cellular stress, were highly upregulated in 17D infected HepG2 cells. The early specific upregulation of these cellular genes in HepG2 cells may be considered markers of the 17D virus. This study on the YF attenuated strain gives a new approach to the analysis of the factors involved in virus attenuation. PMID- 16697682 TI - Effect of microbial pathogens on the diversity of aquatic populations, notably in Europe. AB - The expansion of aquaculture and the demand for ornamental fish have resulted in the large-scale movements of aquatic animals and their pathogens. Here we review the most important non-native fish and shellfish pathogens in European waters and their global impacts on wild fish host populations. The role of theoretical models in the study of the impact of microbial pathogens is discussed, including its integration into risk assessments. PMID- 16697683 TI - Host cell invasion mediated by Trypanosoma cruzi surface molecule gp82 is associated with F-actin disassembly and is inhibited by enteroinvasive Escherichia coli. AB - The target cell F-actin disassembly, induced by a Ca2+-signaling Trypanosoma cruzi factor of unknown molecular identity, has been reported to promote parasite invasion. We investigated whether the metacyclic trypomastigote stage-specific surface molecule gp82, a Ca2+-signal-inducing molecule implicated in host cell invasion, displayed the ability to induce actin cytoskeleton disruption, using a recombinant protein (J18) containing the full-length gp82 sequence fused to GST. J18, but not GST, induced F-actin disassembly in HeLa cells, significantly reducing the number as well as the length of stress fibers. The number of cells with typical stress fibers scored approximately 70% in untreated and GST-treated cells, as opposed to approximately 30% in J18-treated samples, which also showed decreased F-actin content. J18, but not GST, inhibited approximately 6-fold the HeLa cell entry of enteroinvasive Escherichia coli (EIEC), which depends on actin cytoskeleton. Not only were fewer cells infected with bacteria in the presence of J18, there were also fewer bacteria per cell. The inhibitory activity of J18 was Ca2+ dependent. In co-infection experiments, preincubation of HeLa cells with EIEC drastically reduced gp82-dependent internalization of T. cruzi metacyclic forms. All these data, plus the finding that gp82-mediated penetration of metacyclic forms was associated with disrupted HeLa cell cytoskeletal architecture, indicate that gp82 promotes parasite invasion by disassembling the cortical actin cytoskeleton. PMID- 16697684 TI - Brucella spp. lumazine synthase: a novel adjuvant and antigen delivery system to effectively induce oral immunity. AB - Brucella lumazine synthase (BLS) has been previously used with success as a delivery system for systemic immunization against murine cysticercosis. We herein determined the usefulness of BLS as a new antigen-delivery system and mucosal adjuvant using KETc1, one of the peptides of the anti-cysticercosis vaccine. A protection of up to 98% was induced when KETc1 was used as a chimera fused to BLS. Used as adjuvant of KETc1, BLS also induced a high level of protection (79%), which did not significantly differ from that induced by the cholera toxin (74%). KETc1 and BLS administered separately also reduced the parasite load. KETc1 administered orally as a chimera, and to a lesser extent with BLS as adjuvant, elicited IgG and IgA specific antibodies, which were detectable both in fecal extracts and in sera, and increased B and CD4 activated cells. BLS-KETc1 also increased the levels of transcription of TNF-alpha, IL-2 and IFNgamma in Peyer's patches, and in spleen, only increased TNF-alpha was observed. Overall, these results showed that BLS can be used as both an antigen-carrier and as an adjuvant in the design of new oral subunit vaccines. PMID- 16697685 TI - Analysis of gene expression during development: lessons from the Apicomplexa. AB - Apicomplexans are responsible for significant human and animal disease worldwide, including malaria and toxoplasmosis. Herein we summarize recent advances in gene expression analysis in these eukaryotic pathogens, especially with respect to their developmental biology, and discuss the impact this work may have on the development of new vaccines and chemotherapeutics. PMID- 16697686 TI - Differences in gene expression levels and in enzymatic qualities account for the uneven contribution of superoxide dismutases SodCI and SodCII to pathogenicity in Salmonella enterica. AB - Most Salmonella enterica serovars produce two periplasmic [Cu,Zn] superoxide dismutases, SodCI, which is prophage encoded, and SodCII, encoded by a conserved chromosomal gene. Both enzymes were proposed to enhance Salmonella virulence by protecting bacteria against products of macrophage oxidative burst. However, we previously found SodCI, but not SodCII, to play a role during mouse infection by S. enterica serovar Typhimurium. Here we have extended these findings to another serovar of epidemiological relevance: sv Enteritidis. In both serovars, the dominant role of SodCI in virulence correlates with its higher levels in bacteria proliferating in mouse tissues, relative to SodCII. To analyze the basis of these differences, the coding sequences of sodCI and sodCII genes were exchanged with the reciprocal 5'-regions (in serovar Typhimurium). The accumulation patterns of the two proteins in vivo were reversed as a result, indicating that the regulatory determinants lie entirely within the regions upstream from the initiation codon. In the construct with the sodCI gene fused to the sodCII 5' region, SodCI contribution to virulence was reduced but remained significant. Thus, both, high-level expression and some unidentified qualities of the enzyme participate in the phenotypic dominance of SodCI over SodCII in Salmonella pathogenicity. PMID- 16697687 TI - Non-vertebrate hosts in the analysis of host-pathogen interactions. AB - Mutations in bacterial pathogens have been isolated using many strategies. In contrast, the hosts they attack are significantly less tractable. To overcome this problem, a number of model host systems have been developed for isolation and investigation of mutations that modulate pathogen growth. These novel host models are either unicellular organisms, intact invertebrates or cells derived from invertebrates. PMID- 16697688 TI - [Hematocolpos due to obstructed hemivagina. About three cases]. AB - Abnormalities of the reproductive tract interest a little more than 1% of the women. The symptoms are not specific: abdominal pain seems like appendicitis, invalidating dysmenorrheas in the girl, urinary manifestations. They should be discovered and treated because they could bring about obstetric complications later. We present three observations, illustrated with a recent review of the literature, and describe the diagnostic difficulties and their surgical treatments. PMID- 16697689 TI - Illness severity and propensity to travel along the urban-rural continuum. AB - In this paper, we examine whether the relationship between severity of illness and the propensity to travel greater distance relative to the norm (defined by peers in one's county of residence) is uniform across the urban-rural continuum of geography or over time. We focus on the elderly in New York State who have been admitted to hospital for ambulatory care sensitive conditions (ACSCs), admissions which are presumed to be representative of usual travel patterns. The two periods of time examined span the implementation of the Balanced Budget Act (BBA) of 1997, which established the Medicare Rural Hospital Flexibility Program, a major national initiative to strengthen rural health care with the development of rural Critical Access Hospitals (CAHs). As the number of NY rural hospitals certified as CAH increased with the expanded funding from the BBA, one might expect to see increased distance traveled by more severely ill rural elderly, as their CAHs referred them to their affiliated support hospitals. The logistic regression estimates support this expectation, highlighting an asymmetrical relationship between relative distance and severity across patients in rural and urban areas. Despite a general decline in average propensity to travel further than the norm across the landscape, severity had a larger impact on travel propensity in rural areas, which increased over time. PMID- 16697690 TI - Induction of interleukin-8 secretion and activation of ERK1/2, p38 MAPK signaling pathways by thrombin in dermal fibroblasts. AB - It was reported that thrombin could induce IL-8 secretion from human dermal fibroblasts (HDFs) through activation of proteinase activated receptor (PAR)-1. However, little is known of intracellular signaling pathways involved in the event. In the present study, expression of PARs in primarily cultured HDFs was determined by flow cytometry analysis and reverse transcription polymerase chain reaction (RT-PCR), levels of IL-8 were determined by using ELISA and signaling pathways were examined by using Western blot. It was found that HDFs express PAR 1 and PAR-3, and thrombin induces approximately 7.4-fold increase in IL-8 secretion from HDFs. Hirudin and a PAR-1 blocking antibody completely abolish the action of thrombin. It was also found that PD98059, a mitogen-activated protein kinase (MAPK) pathway inhibitor and U0126, an inhibitor of extracellular signal regulated kinase (ERK) blocks thrombin-induced phosphorylation of ERK1/2 and IL-8 secretion, indicating the involvement of MAPK/ERK signaling pathway in thrombin induced IL-8 secretion. p38 MAPK pathway appears also being involved as SB203580, a selective inhibitor of p38 MAPK inhibit phosphorylation of p38 MAPK and thrombin-induced IL-8 secretion. Furthermore, Janus kinase (JAK)/signal transducer and activator of transcription 3 (STAT3) pathway, but not phosphotidylinositol 3-kinase (PI3K)/Akt signaling pathway may also be activated by thrombin. In conclusion, thrombin potently induce IL-8 release via PAR-1 from HDFs. Thrombin elicited IL-8 release is predominantly conducted through MAPK/ERK and p38 MAPK signaling pathways. Discovery of the signaling pathways of thrombin in HDFs may help to understand the role of thrombin in inflammation and tissue remodeling. PMID- 16697691 TI - Skeletal muscle apoptosis is not increased in gastric cancer patients with mild moderate weight loss. AB - Numerous experimental and clinical studies have shown that skeletal muscle apoptotis may increase in wasting conditions and suggest that apoptosis might contribute to the loss of lean body mass. Data in cancer patients are still lacking. The present study aimed at verifying whether apoptosis was enhanced in the skeletal muscle of 16 patients with gastric cancer with respect to controls. A biopsy specimen was obtained from the rectus abdominis muscle. The occurrence of apoptosis in muscle biopsies was determined morphologically by the fluorescent transferase-mediated dUTP nick end labeling assay and by immunohistochemistry for caspase-3 and caspase-1. Mean weight loss was 6+/-2% in cancer patients and 0.5+/ 0.1% in controls (p<0.0001). Serum albumin levels (g/dL) were 3.7+/-0.3 in cancer patients and 4.1+/-0.2 in controls (p<0.05). The percentage of apoptotic myonuclei was similar in cancer patients and in controls (1.5+/-0.3 versus 1.4+/ 0.2, respectively; p=ns), in gastric cancer patients with mild (1.6+/-0.4) or moderate-severe weight loss (1.4+/-0.5) (p=ns), and in the different stages of disease (stages I-II: 1.5+/-0.7; stage III: 1.3+/-0.4; stage IV: 1.6+/-0.3; p=ns). By immunohistochemistry, caspase-1 and caspase-3 positive fibers were absent in controls and in neoplastic patients. Poly-ADP-ribosyl polymerase, a typical caspase-3 substrate whose processing is indicative of caspase-3 activation, was not cleaved in muscle biopsies of cancer patients. These data suggest that skeletal muscle apoptosis is not increased in neoplastic patients with mild-moderate weight loss and argue against the hypotheses that caspase-3 activation might be an essential step of myofibrillar proteolysis in cancer related muscle wasting. PMID- 16697692 TI - Heme oxygenase-1 inhibits apoptosis in Caco-2 cells via activation of Akt pathway. AB - Heme oxygenase-1 can play a protective role against cellular stress. In colon cancer cells, these effects would be relevant to oncogenesis and resistance to chemotherapy. The aim of the study was to examine the effects of heme oxygenase-1 induction on cell survival in a human colon cancer cell line, Caco-2. Serum deprivation induced apoptosis, reduced Akt and p38 phosphorylation, and increased p21(Cip/WAF1) levels. Heme oxygenase-1 induction by treatment with cobalt protoporphyrin IX resulted in resistance to apoptosis, activation of Akt, reduction in p21(Cip/WAF1) levels and modification of bcl2/bax ratio towards survival. Indomethacin reduced apoptosis but in contrast to heme oxygenase-1, arrested cells in G0/G1. Apoptosis was also inhibited by the heme oxygenase metabolites bilirubin and biliverdin but the CO donor tricarbonyldichlororuthenium(II) dimer did not exert significant effects. Protection against apoptosis in cells treated with cobalt protoporphyrin IX was reverted by incubation with heme oxygenase-1 small interfering RNA. This study shows an antiapoptotic effect of heme oxygenase-1 in colon cancer cells which could be mediated by the formation of bilirubin and biliverdin. Our results support an antiapoptotic role for HO-1 in these cells and provide a mechanism by which overexpression of HO-1 may promote tumor resistance to stress in conditions of limited nutrient supply. We have extended these observations by demonstrating that these effects are independent of p38 but are mediated via Akt pathway. PMID- 16697693 TI - Can metals defend plants against biotic stress? AB - Farmers have used metal compounds in phytosanitary treatments for more than a century; however, it has recently been suggested that plants absorb high concentrations of metals from the substrate as a self-defense mechanism against pathogens and herbivores. This metal defense hypothesis is among the most attractive proposals for the 'reason to be' of metal hyperaccumulator species. On a molecular basis, metal defense against biotic stress seems to imply common and/or complementary pathways of signal perception, signal transduction and metabolism. This does not imply a broad band of co-resistance to different stress types but reflects a continuous cross talk during the coevolution of plants, pathogens and herbivores competing in an environment where efficient metal ion acquisition and ion homeostasis are essential for survival. PMID- 16697694 TI - Native-language sensitivities: evolution in the first year of life. AB - In the first few months of their lives, human infants can perceive differences in phonetic contrasts not present in their environment and that their parents cannot discriminate. At 12 months, their pattern of discrimination approaches that of their parents. Until recently, it was believed that the perception of native language categories did not significantly evolve in that period. Kuhl et al. offer compelling new evidence indicating significant increase in native-language perception. PMID- 16697695 TI - Cognitive abnormalities in post-traumatic stress disorder. AB - Characteristically arising in response to overwhelmingly terrifying events, post traumatic stress disorder (PTSD) is a disorder of memory: sufferers seemingly relive their trauma in the form of involuntary recollection. Prominent cognitive abnormalities, especially in memory functioning, have motivated research designed to elucidate the mediating mechanisms that produce PTSD symptoms, especially those involving involuntary recollection. Recent developments suggest a pathophysiological model of PTSD which includes hyporesponsive prefrontal cortical regions and/or a hyper-responsive amygdala. Other work has also identified above-average cognitive ability as a protective factor and below average hippocampal volume as a vulnerability factor for PTSD among the trauma exposed. These attempts to elucidate the mediating mechanisms of PTSD have been both cognitive and, more recently, cognitive-neuroscientific in emphasis. PMID- 16697696 TI - Ultraviolet (UV) transmittance characteristics of daily disposable and silicone hydrogel contact lenses. AB - The ultraviolet (UV) transmittance spectra of daily wear hydrogel and disposable silicone hydrogel contact lenses were measured. Average transmittance percentages were calculated for each lens for the entire UV spectrum and individually for the UVC, UVB and UVA portions of the spectrum. The significance of the differences in transmittance spectra obtained for the lenses was analysed using a one-way ANOVA planned comparisons test (alpha=0.05). The transmittance data were then used to calculate a UV protection factor (PF) for each contact lens brand tested. The PFs for 1-DAY ACUVUE MOIST (6.22), ACUVUE ADVANCE (10.02) and ACUVUE OASYS (11.96) contact lenses show that these contact lenses have superior UV-blocking capabilities. The PFs for Focus DAILIES (1.79), SofLens 1-day disposables (1.72), NIGHT & DAY (1.84), O2 Optix (1.99) and Purevision (2.62) show that these contact lenses posses more modest UV-blocking characteristics. This paper reviews the importance of protection of the anterior ocular surface from UV damage and quantifies the protection afforded by selected commercially available disposable contact lenses. PMID- 16697697 TI - A simple variational-numerical approach to the ro-vibrational spectrum of diatomic molecules. An application to CH+. AB - We describe a simple way of obtaining numerically the manifold of energies for ro vibrational transitions for a centrifugally distorted oscillator, starting from the potential energy of the non-rotating oscillator calculated by an accurate ab initio method. It is shown that the energies so obtained compare well with those obtained variationally. The species of astrophysical interest methylidyne ion, CH(+), has been selected as an example that allow us to show the computational efficiency of the method with respect to the variational one. It is applied for the determination of ro-vibrational levels up J=6, and the spectroscopic parameters corresponding to the ground electronic state X(1)Sigma(+). From the potential energy surface computed at the MRCI/cc-pV5Z level, the fundamental frequency, B(0) and D(0) are determined to be 2724.8, 13.85688 and -1.53322x10( 3)cm(-1), respectively. We provide also an estimation of anharmonic constants. PMID- 16697698 TI - Cytomegalovirus and Epstein-Barr virus subtypes--the search for clinical significance. AB - Cytomegalovirus (CMV) as well as Epstein Barr virus (EBV) genomes include regions which show in part substantial polymorphisms. Characterization of several polymorphic regions led to the identification of various CMV and EBV subtypes. Within the last years there have been undertaken numerous efforts to find out whether the diverse subtypes differentially contribute to clinical manifestations. However, although some associations have been described so far between a certain virus subtype and the development of individual diseases these analyses were greatly complicated by the huge genomic background of CMV and EBV, by the large variety of individual host-virus relations and by differences in the geographic or demographic subtype distribution. In addition, it was shown meanwhile that a substantial proportion of virus infections is due to mixed infections with different subtypes. In this review we will give an overview of the current knowledge concerning the clinical significance of individual CMV and EBV subtypes, defined by characterization of selected polymorphisms. In addition, we also focus on recent analyses which show that infection with mixed virus subtype populations may be disadvantageous compared to single virus subtype infections. PMID- 16697699 TI - Hypothyroidism down-regulates mitochondrial citrate carrier activity and expression in rat liver. AB - The effect of hypothyroidism on citrate carrier (CiC) activity has been investigated in rat-liver mitochondria. The rate of citrate transport was reduced by approximately 50% in mitochondria from hypothyroid as compared with euthyroid rats. In parallel, a decrease in the rate of de novo fatty acid synthesis was observed in the cytosol of the former animals. Kinetic analysis of citrate transport revealed that only the Vmax was reduced by hypothyroidism, while Km was almost unaffected. Hypothyroidism increased the mitochondrial percentage of phosphatidylcholine while decreased that of phosphatidylethanolamine; an altered fatty acid pattern but no significant difference in the sum of saturated and unsaturated fatty acids as well as in the unsaturation index was observed. The CiC Arrhenius plot did not show appreciable difference between the two groups of rats. However, Western blot analysis associated with mRNA quantitation indicated that both protein level and mRNA accumulation of hepatic CiC were noticeably decreased in hypothyroid state. Therefore, a reduced content of the carrier protein can represent a plausible mechanism to explain the decline in the CiC activity observed in rat liver mitochondria of hypothyroid rats. PMID- 16697700 TI - Adaptation of office workers to a new building - impaired well-being as part of the sick-building-syndrome. AB - AIM: The focus of our study was the assessment of the effects of spatial relocation on office staff. Our aim was to investigate whether psychosocial or personal factors are better predictors of the occurrence of impaired well-being. METHODS: Before relocation the administration of the university hospital of Ulm (Germany) was located in ten different buildings. Chemical and physical parameters of the indoor air were measured. The employees were surveyed with a questionnaire for their health status and psychosocial determinants. After moving to a new wide-spaced building, the same procedure was reapplied shortly afterwards and half a year later. Only respondents who had taken part in all three surveys are taken into account (n=84). The definition of impaired well being as defined by the ProKlimA-study group was used as the criterion variable. RESULTS: The overall prevalence of impaired well-being rose from 24% to 36% after relocation. Contrarily, persons who were formerly accommodated in a wide spaced building showed a reduced risk (OR(post1)=0.3). Affected persons had at all times a more negative response pattern. Chemical and physical parameters did not have any influence in this context. CONCLUSIONS: The adaptation to a new environment is influenced by the old "socialization" of the former buildings. Impaired well being is not limited to bodily complaints, it rather has a systemic character in the form of a distinctive overall response pattern. For an adequate analysis of impaired well-being - and the sick-building-syndrome in consequence - the elucidation of individual and other potentially intervening factors is essential. Taking this into consideration, the search for norm values or a framework seems to be of limited value. PMID- 16697701 TI - Critical differences in lateral X-rays with and without a diagnosis of plantar fasciitis. AB - Plantar fasciitis is a clinical diagnosis and is often combined with some form of imaging to validate the diagnosis. The clinical utility of lateral X-rays lies in the fact that they are relatively inexpensive and may contribute to ruling out other osseous causes of pain. In this study 106 (27 plantar fasciitis (PF) and 79 controls) plain non-weight bearing lateral X-rays were examined by a blind examiner to document the key features of the lateral X-ray between images of individuals with and without plantar fasciitis. As expected calcaneal spurs were observed in both groups (85% PF and 46% controls). However, plantar fascia thickness and fat pad abnormalities resulted in the best group differentiation (p<0.0001) with sensitivity of 85% and specificity of 95% for plantar fasciitis. It was concluded that the key radiological features that differentiate the groups were not spurs but rather changes in the soft tissues. If it is deemed necessary to confirm the diagnosis of typical plantar fasciitis with imaging, a lateral non weight bearing X-ray should be the first choice investigation especially if these key features are noted. PMID- 16697702 TI - Assessment of bone strength at differentially-loaded skeletal regions in adolescent middle-distance runners. AB - Bone adaptations to loading extend beyond mineral accrual to geometric markers of bone strength. Available technology and regional differences in cortical bone dictate how bone strength is reported. Examination of bone strength at two differentially-loaded skeletal sites using hip structure analysis (HSA) and bone strength index (BSI) is under-explored in adolescent sporting populations. The purpose of this study was to compare HSA at the femoral neck and BSI at the distal tibia in adolescent middle-distance runners and age- and gender-matched controls. Four groups of 20 adolescents aged 14-18 years were composed of male and female middle-distance runners, and male and female controls. Distal tibial BSI was calculated using data from dual energy X-ray absorptiometry (DXA) and magnetic resonance imaging (MRI). Calculations for femoral neck strength were acquired from DXA-derived HSA software. Female athletes displayed greater distal tibial BSI than controls t(38)=3.4, p=0.002, but femoral neck bone measures did not differ. In males, no group differences were found at either the distal tibia or femoral neck. In conclusion, exposure to similar high training loads may advantage female adolescent athletes more than male adolescent athletes compared with less active peers in bone strength at the distal tibia. PMID- 16697703 TI - Minimally invasive approach for surgical management of cardiac myxoma. PMID- 16697704 TI - Rotational atherectomy and simultaneous kissing stenting of a bifurcation stenosis of left anterior descending and anomalous right coronary arteries. AB - Intervention to anomalous coronaries can present specific challenges: non standard guide catheters may be required to provide adequate access and support, the course of the arteries can be tortuous with sharp angulations, and they may subtend a critical area of myocardium. We present the first report of percutaneous coronary intervention to the bifurcation of an anomalous right coronary artery arising from the left anterior descending artery. This lesion contained a high burden of calcification, which required rotational atherectomy of the left anterior descending artery before kissing stenting of the bifurcation. PMID- 16697705 TI - Acute coronary syndrome induced by capecitabine therapy. AB - Capecitabine, a new member of the fluoropyrimidine family, is an orally administered drug that delivers fluorouracil (5-FU) selectively to the tumour. Although the cardiotoxicity of 5-FU is well documented, there is little published data about cardiovascular adverse effects of Capecitabine. This case highlights the possible development of acute coronary syndrome as a side effect of Capecitabine therapy. PMID- 16697706 TI - T cell recognition in chronic beryllium disease. AB - Chronic beryllium disease (CBD) is a granulomatous lung disorder caused by beryllium exposure in the workplace and is characterized by the accumulation of beryllium-specific CD4(+) T cells. Depending on genetic susceptibility and the nature of the exposure, CBD occurs in up to 20% of exposed workers. Genetic susceptibility has been associated with particular HLA-DP alleles, especially those possessing a negatively charged glutamic acid residue at the 69th position of the beta-chain. The mechanism for this association lies in the ability of these HLA-DP molecules to bind and present beryllium to pathogenic CD4(+) T cells. Large numbers of effector memory, beryllium-specific CD4(+) T cells are recruited to the lung of these subjects and secrete Th1-type cytokines upon beryllium recognition. The presence of circulating beryllium-specific CD4(+) T cells directly correlates with the severity of lymphocytic alveolitis. With the presence of a known antigenic stimulus, CBD serves as an important model of immune-mediated, organ destruction. Thus, our findings in CBD have important implications for studies in autoimmune diseases, in particular those with an unknown inciting antigen and an inaccessible target organ. PMID- 16697707 TI - Development of novel E1-complementary cells for adenoviral production free of replication-competent adenovirus. PMID- 16697708 TI - A conditionally replicating HIV-based vector that stably expresses an antiviral shRNA against HIV-1 replication. AB - Human pathogenic viruses can be targeted by therapeutic strategies based on RNA interference. Whereas the administration of synthetic short interfering RNAs (siRNAs) may transiently inhibit viral replication, long-term inhibition may be achieved through stable intracellular expression of siRNAs or short hairpin RNAs (shRNAs). Both approaches face serious problems with delivery to the right cells in an infected individual. We explored the potential of a replicating HIV-based vector to deliver an antiviral shRNA cassette into HIV-1-susceptible target cells to block chronic HIV-1 infection. The vector is based on a doxycycline (dox) dependent HIV-1 variant that we previously proposed as a conditional-live HIV-1 vaccine. With dox, this virus spreads efficiently to all HIV-susceptible cells. Subsequent dox withdrawal generates cells with a transcriptionally silent integrated provirus, but with an active shRNA expression cassette. Because the shRNA targets viral sequences that are removed from the vector construct, there is no self-targeting, yet there is specific shutdown of HIV-1 replication. PMID- 16697709 TI - Aphasic status epilepticus with periodic lateralized epileptiform discharges in a bilingual patient as a presenting sign of "AIDS-toxoplasmosis complex". AB - We describe an HIV-infected, bilingual patient presenting with Wernicke's aphasia due to partial status epilepticus with periodic lateralized epileptiform discharges, as the first sign of AIDS-toxoplasmosis complex. The localization of the native and secondary language centers in the brain and the possible role of recurrent seizures in the fluctuating course of Wernicke's aphasia in this patient are discussed. The clinical course of this patient supports the belief that a second language area for a second language learned in the later stages of life is located in an area different from that for the native language but still in close proximity to it. PMID- 16697710 TI - NCI Thesaurus: a semantic model integrating cancer-related clinical and molecular information. AB - Over the last 8 years, the National Cancer Institute (NCI) has launched a major effort to integrate molecular and clinical cancer-related information within a unified biomedical informatics framework, with controlled terminology as its foundational layer. The NCI Thesaurus is the reference terminology underpinning these efforts. It is designed to meet the growing need for accurate, comprehensive, and shared terminology, covering topics including: cancers, findings, drugs, therapies, anatomy, genes, pathways, cellular and subcellular processes, proteins, and experimental organisms. The NCI Thesaurus provides a partial model of how these things relate to each other, responding to actual user needs and implemented in a deductive logic framework that can help maintain the integrity and extend the informational power of what is provided. This paper presents the semantic model for cancer diseases and its uses in integrating clinical and molecular knowledge, more briefly examines the models and uses for drug, biochemical pathway, and mouse terminology, and discusses limits of the current approach and directions for future work. PMID- 16697711 TI - Neurogenesis and neural stem cells in the dorsal vagal complex of adult rat brain: new vistas about autonomic regulations--a review. AB - The dorsal vagal complex (DVC) of the brainstem is the major reflex center of autonomic nervous system. Several neuroplasticity effectors have been identified in the DVC of adult rat, such as PSA-NCAM, GAP-43, BDNF and its receptor TrkB; moreover, acute vagal stimulation was found to induce c-fos and to down-regulate western-blot-assayed tissular concentration of PSA-NCAM. Adult neurogenesis was first shown in rat DVC by BrdU incorporation combined with phenotypic labelling in situ; new neurons are generated in equal proportions with new astrocytes and at a lower rate than in olfactory bulb or hippocampus. Intrinsic proliferative cells were then detected within the DVC of adult rat by means of Ki-67 immunohistochemistry and western-blot of D-cyclins. The presence of neural stem cells within DVC was directly demonstrated by applying the in vitro neurosphere assay on microdissected adult DVC explants; DVC-derived neurospheres display lower proliferation rate and neurogenic potential than forebrain ones. Vagotomy in adult promotes massive and transient increase of neurogenic and microglial proliferations within DVC, the kinetics and location of which were analyzed by Ki 67 immunohistochemistry and cyclin D western blot. These mechanisms shed light on so far unknown plasticity potential in DVC, which brings novel cues about physiological adaptations of autonomic reflexes in adult mammals. PMID- 16697712 TI - Cerebral and muscle tissue oxygenation in acute hypoxic ventilatory response test. AB - Eight men were exposed to progressive isocapnic hypoxia for 10 min to test the hypothesis that (i) cerebral and muscle tissue would follow similar deoxygenation profiles during an acute hypoxic ventilatory response (AHVR) test; and (ii) strong cerebrovascular responsiveness to hypoxia would be related to attenuated cerebral deoxygenation. End-tidal O(2) concentration was reduced from normoxia (approximately 102 mmHg) to approximately 45 mmHg while arterial oxygen saturation (SpO2 %) declined from 98+/-1% to 77+/-7% (P<0.001). Near-infrared spectroscopy (NIRS)-derived local cerebral tissue (frontal lobe) deoxyhemoglobin increased 5.55+/-2.22 microM, while oxyhemoglobin and tissue oxygenation index decreased 2.57+/-1.99 microM and 6.2+/-3.4%, respectively (all P<0.001). In muscle (m. vastus lateralis) the NIRS changes from the initial normoxic level were non-significant. Cerebral blood velocity (V(mean), transcranial Doppler) in the middle cerebral artery increased from 53.4+/-10.4 to 60.6+/-11.6 cms(-1) (P<0.001). In relation to the decline in SpO2 % the mean rate of increase of V(mean) and AHVR were 0.33+/-0.19 cms(-1)%(-1) and 0.52+/-0.20l min(-1)%(-1), respectively. We conclude that cerebral, but not muscle, tissue shows changes reflecting a greater deoxygenation during acute hypoxia. However, the changes in NIRS parameters were not related to cerebrovascular responsiveness or ventilatory chemosensitivity during graded hypoxia. PMID- 16697716 TI - CMS selects demonstration sites for ways to reduce disparities in cancer health care. PMID- 16697713 TI - Bisphosphonates activate nucleotide receptors signaling and induce the expression of Hsp90 in osteoblast-like cell lines. AB - Bisphosphonates are the most important drugs used in the treatment of osteoporosis as they inhibit osteoclast resorption and stimulate proliferation of osteoblasts. However, the molecular mechanisms responsible for these effects are still poorly elucidated. It is known that nucleotide receptors-mediated signaling plays a central role in modulating osteoblasts growth in response to mechanical stress. By using osteoblast-like cell lines (i.e., HOBIT, MG-63, ROS P2Y), which express P2Y receptors, we found that the treatment with risedronate promotes non lytic ATP release leading to activation of ERKs through the involvement of P2Y receptors triggering. A major role in this signal transduction pathway seems to be the involvement of P2Y(1) and P2Y(2) receptors, since the stimulatory effect of risedronate on ERKs is not appreciable in ROS 17/2.8 cells, which do not express these two receptors. Differential proteomics analysis identified Hsp90 upregulation as a result of risedronate effect on HOBIT and MG-63 cells. The stimulatory effect is dependent on ERKs activation involving nucleotide receptors triggering and leads to increased proliferation of osteoblast-like cells. In fact, functional inactivation of Hsp90 by the specific inhibitor 17-AAG prevents the bisphosphonate-induced mitogenic effects in osteoblasts. These findings show that bisphosphonates, by inducing ATP release, may also act through nucleotide receptors signaling leading to ERKs activation and may exert their mitogenic role on osteoblasts through the involvement of Hsp90. PMID- 16697717 TI - CMS expands opportunities for physicians to earn quality improvement performance payments. PMID- 16697718 TI - The future of gastroenterology and gastroenterology. PMID- 16697719 TI - Gastric bypass surgery improves metabolic and hepatic abnormalities associated with nonalcoholic fatty liver disease. AB - BACKGROUND & AIMS: Most patients with extreme obesity have nonalcoholic fatty liver disease (NAFLD). Although gastric bypass (GBP) surgery is the most common bariatric operation performed in obese patients in the United States, the effect of GBP surgery-induced weight loss on the metabolic and hepatic abnormalities associated with NAFLD are not clear. METHODS: Whole-body glucose, fatty acid and lipoprotein kinetics, liver histology, and hepatic cellular factors involved in inflammation and fibrogenesis were evaluated in 7 extremely obese subjects (body mass index, 58 +/- 4 kg/m(2)) before and 1 year after GBP surgery. RESULTS: At 1 year after surgery, subjects lost 29% +/- 5% of initial body weight (P < .01); palmitate rate of appearance in plasma, an index of adipose tissue lipolysis, decreased by 47% +/- 4% (P < .01); endogenous glucose production rate decreased by 27% +/- 7% (P < .01); and very-low-density lipoprotein-triglyceride secretion rate decreased by 44% +/- 9% (P < .05). In addition, GBP surgery-induced weight loss decreased hepatic steatosis but did not change standard histologic assessments of inflammation and fibrosis. However, there was a marked decrease in hepatic factors involved in regulating fibrogenesis (collagen-alpha1(I), transforming growth factor-beta1, alpha-smooth muscle actin, and tissue inhibitor of metalloproteinase 1 expression and alpha-smooth muscle actin content) and inflammation (macrophage chemoattractant protein 1 and interleukin 8 expression) (P < .05, compared with values before weight loss). CONCLUSIONS: These data demonstrate that weight loss induced by GBP surgery normalizes the metabolic abnormalities involved in the pathogenesis and pathophysiology of NAFLD and decreases the hepatic expression of factors involved in the progression of liver inflammation and fibrosis. PMID- 16697720 TI - Gastrointestinal stromal tumors with KIT exon 11 deletions are associated with poor prognosis. AB - BACKGROUND & AIMS: Gain-of-function mutations in the KIT receptor tyrosine kinase gene and rare mutations in the platelet-derived growth factor receptor alpha (PDGFRA) gene are important events in gastrointestinal stromal tumor (GIST) development. Different mutations are reportedly associated with distinctive phenotypes and possibly clinical behavior. We investigated the correlation among mutation type, phenotype, and clinical course in a preimatinib, population-based series of GIST with long-term follow-up. METHODS: Genomic DNA from 177 GIST patients was analyzed for KIT exons 9, 11, 13, and 17 and PDGFRA exons 12 and 18 mutations using denaturating high-performance liquid chromatography and bidirectional sequencing. RESULTS: KIT exon 11 mutations were detected in 101 of 177 GIST (61 deletions, 23 missense mutations, and 17 duplications); wild-type (WT) KIT and PDGFRA were detected in 63; KIT exon 9 and exon 17 mutations in 6 and 1, respectively; and PDGFRA exons 12 and 18 mutations in 3 each. GIST >5 cm vs GIST /=2) at 3 months (75% vs 41%; odds ratio, 4.36; 95% confidence interval, 1.44 13.23; P = .008) and at 1 year (93% vs 59%; odds ratio, 9.80; 95% confidence interval, 1.04-92.70; P = .041) and had a trend toward an earlier reintervention (mean, 7.00 vs 5.30 years; P = .174). The severity of myenteric plexitis in the proximal resection margin correlated with the severity of endoscopic recurrence at 3 months (r = 0.334, P = .010) and 1 year (r = 0.560, P = .001). Myenteric plexitis was the only consistent predictor of endoscopic recurrence. CONCLUSIONS: The presence of myenteric plexitis in proximal margins of ileocolonic resection specimens is predictive of early endoscopic CD recurrence. PMID- 16697724 TI - Alcohol use and treatment of hepatitis C virus: results of a national multicenter study. AB - BACKGROUND & AIMS: Patients with hepatitis C virus (HCV) infection who use alcohol have been excluded from clinical trials; therefore, outcomes with antiviral therapy are unknown. The aim of the study was to determine the impact of alcohol use on HCV treatment outcomes. METHODS: Subjects using alcohol were categorized as follows: no alcohol versus regular alcohol use, quantity consumed (none, <6 drinks/day, >/=6 drinks/day), CAGE score <2 or >/=2, and recent alcohol use (past 12 months). Patients were treated with interferon plus ribavirin. RESULTS: A total of 4061 subjects were enrolled, and 726 (18%) received treatment. Alcohol use (past and within 12 months) reduced treatment candidacy. Past alcohol use did not affect the end-of-treatment response, sustained virologic response (SVR), and treatment discontinuation rates. However, recent alcohol use resulted in higher treatment discontinuation (40% vs 26%; P = .0002) and tended to reduce the SVR (14% vs 20%; P = .06), but when patients who discontinued treatment were excluded from analysis, the trend in favor of nondrinkers for SVR disappeared (25% vs 23%). These findings were also consistent in subgroup analyses on race and genotype. CONCLUSIONS: Eligibility for anti-HCV treatment was reduced in past and recent drinkers. Recent alcohol use was associated with increased treatment discontinuation and lower SVR. However, patients who use alcohol and completed the treatment had a response comparable to that of nondrinkers. Patients with a history of alcohol use should not be excluded from HCV therapy. Instead, additional support should be provided to these patients to ensure their ability to complete treatment. PMID- 16697725 TI - The evolution of severe steatosis after bariatric surgery is related to insulin resistance. AB - BACKGROUND & AIMS: In severely obese patients, factors implicated in the evolution of severe steatosis after bariatric surgery remain unresolved. Our aim was to determine whether insulin resistance (IR) influences the histologic effects induced by bariatric surgery. METHODS: We prospectively included 185 severely obese patients (body mass index >/=35 kg/m(2)) referred for bariatric surgery. The evolution of IR (IR index = 1/quantitative insulin sensitivity check index) and liver injury with consecutive biopsy was concomitantly assessed before and 1 year after surgery. RESULTS: At preoperative biopsy, 27% of severely obese patients disclosed severe steatosis (>/=60%). The alanine aminotransferase (P = .01) and IR indexes (P = .04) were independent predictive factors of severe steatosis at baseline. One year after surgery, surgical treatment induced a decrease in body mass index (9.5 kg/m(2); P < .0001), steatosis score (8.5%; P < .0001), and IR index (0.29; P < .0001). The preoperative IR index (P = .01) and preoperative steatosis (P = .006) were independent predictive factors in the persistence of severe steatosis after surgery. Moderate or severe steatosis was more frequently observed in patients who had conserved a higher IR index after surgery than in patients who had improved their IR index (44% vs 20.2%; P = .04). CONCLUSIONS: IR was independently associated with severe steatosis and predicted its persistence after surgery. The amelioration of IR after surgery is associated with a decrease in the amount of fat. Taken together, the results of this prospective study in severely obese patients demonstrate that severe steatosis and its evolution after surgery are intimately connected with IR. PMID- 16697726 TI - Genotypes and viremia of hepatitis B and D viruses are associated with outcomes of chronic hepatitis D patients. AB - BACKGROUND & AIMS: Genotypes and viremia of hepatitis D virus (HDV) and hepatitis B virus (HBV) may be associated with outcomes. This study evaluated the impact of viral genotypes and viremia on outcomes of dual HBV and HDV infection. METHODS: Viremia and viral genotypes were analyzed in 194 consecutive chronic hepatitis B patients with HDV superinfection and correlated with outcomes. RESULTS: The numbers of HBV genotype A, B, C, and nonclassified were 4, 57, 23, and 110, respectively. There were 51 genotype I HDV, 74 genotype II HDV, 8 genotype IV HDV, and 61 nonclassified HDV genotype. In a median follow-up of 135 months, 24 progressed to cirrhosis and 41 developed hepatocellular carcinoma. Patients infected with genotype I HDV had a lower remission rate (15.2% vs 40.2%; P = .007) and more adverse outcomes (cirrhosis, hepatocellular carcinoma, or mortality) (52.2% vs 25.0%; P= .005) than those with genotype II HDV. Patients infected with genotype C HBV had a lower remission rate (0 vs 32.1%; P = .005) and more adverse outcomes (70.0% vs 33.9%; P = .005) than those with genotype B HBV. The presence of HBV or HDV viremia was associated with lower remission rates compared with those negative for both (26.4% and 24.3% vs 69.2%; P < .001). In multivariate analysis, age, genotype C HBV, and genotype I HDV were independent factors associated with adverse outcomes. CONCLUSIONS: In chronic HBV and HDV dual infections, older age, genotype I HDV, and genotype C HBV correlated with adverse outcomes. PMID- 16697727 TI - Relationship between steatosis, inflammation, and fibrosis in chronic hepatitis C: a meta-analysis of individual patient data. AB - BACKGROUND & AIMS: Steatosis is a frequent histologic finding in chronic hepatitis C (CHC), but it is unclear whether steatosis is an independent predictor for liver fibrosis. We evaluated the association between steatosis and fibrosis and their common correlates in persons with CHC and in subgroup analyses according to hepatitis C virus (HCV) genotype and body mass index. METHODS: We conducted a meta-analysis on individual data from 3068 patients with histologically confirmed CHC recruited from 10 clinical centers in Italy, Switzerland, France, Australia, and the United States. RESULTS: Steatosis was present in 1561 patients (50.9%) and fibrosis in 2688 (87.6%). HCV genotype was 1 in 1694 cases (55.2%), 2 in 563 (18.4%), 3 in 669 (21.8%), and 4 in 142 (4.6%). By stepwise logistic regression, steatosis was associated independently with genotype 3, the presence of fibrosis, diabetes, hepatic inflammation, ongoing alcohol abuse, higher body mass index, and older age. Fibrosis was associated independently with inflammatory activity, steatosis, male sex, and older age, whereas HCV genotype 2 was associated with reduced fibrosis. In the subgroup analyses, the association between steatosis and fibrosis invariably was dependent on a simultaneous association between steatosis and hepatic inflammation. CONCLUSIONS: In this large and geographically different group of CHC patients, steatosis is confirmed as significantly and independently associated with fibrosis in CHC. Hepatic inflammation may mediate fibrogenesis in patients with liver steatosis. Control of metabolic factors (such as overweight, via lifestyle adjustments) appears important in the management of CHC. PMID- 16697728 TI - Distal splenorenal shunt versus transjugular intrahepatic portal systematic shunt for variceal bleeding: a randomized trial. AB - BACKGROUND & AIMS: Variceal bleeding refractory to medical treatment with beta blockers and endoscopic therapy can be managed by variceal decompression with either surgical shunts or transjugular intrahepatic portal systemic shunts (TIPS). This prospective randomized trial tested the hypothesis that patients receiving distal splenorenal shunts (DSRS) would have significantly lower rebleeding and encephalopathy rates than TIPS in management of refractory variceal bleeding. METHODS: A prospective randomized controlled clinical trial at 5 centers was conducted. One hundred forty patients with Child-Pugh class A and B cirrhosis and refractory variceal bleeding were randomized to DSRS or TIPS. Protocol and event follow-up for 2-8 years (mean, 46 +/- 26 months) for primary end points of variceal bleeding and encephalopathy and secondary end points of death, ascites, thrombosis and stenosis, liver function, need for transplant, quality of life, and cost were evaluated. RESULTS: There was no significant difference in rebleeding (DSRS, 5.5%; TIPS, 10.5%; P = .29) or first encephalopathy event (DSRS, 50%; TIPS, 50%). Survival at 2 and 5 years (DSRS, 81% and 62%; TIPS, 88% and 61%, respectively) were not significantly different (P = .87). Thrombosis, stenosis, and reintervention rates (DSRS, 11%; TIPS, 82%) were significantly (P < .001) higher in the TIPS group. Ascites, need for transplant, quality of life, and costs were not significantly different. CONCLUSIONS: DSRS and TIPS are similarly efficacious in the control of refractory variceal bleeding in Child-Pugh class A and B patients. Reintervention is significantly greater for TIPS compared with DSRS. Because both procedures have equivalent outcomes, the choice is dependent on available expertise and ability to monitor the shunt and reintervene when needed. PMID- 16697729 TI - Evidence-based incorporation of serum sodium concentration into MELD. AB - BACKGROUND & AIMS: Serum sodium (Na) concentrations have been suggested as a useful predictor of mortality in patients with end-stage liver disease awaiting liver transplantation. METHODS: We evaluated methods to incorporate Na into model for end-stage liver disease (MELD), using a prospective, multicenter database specifically created for validation and refinement of MELD. Adult, primary liver transplant candidates with end-stage liver disease were enrolled. RESULTS: Complete data were available in 753 patients, in whom the median MELD score was 10.8 and sodium was 137 mEq/L. Low Na (<130 mEq/L) was present in 8% of patients, of whom 90% had ascites. During the study period, 67 patients (9%) died, 243 (32%) underwent transplantation, 73 (10%) were withdrawn, and 370 were still waiting. MELD score and Na, at listing, were significant (both, P < .01) predictors of death within 6 months. After adjustment for MELD score and center, there was a linear increase in the risk of death as Na decreased between 135 and 120 mEq/L. A new score to incorporate Na into MELD was developed: "MELD-Na" = MELD + 1.59 (135 - Na) with maximum and minimum Na of 135 and 120 mEq/L, respectively. In this cohort, "MELD-Na" scores of 20, 30, and 40 were associated with 6%, 16%, and 37% of risk of death within 6 months of listing, respectively. If this new score were used to allocate grafts, it would affect 27% of the transplant recipients. CONCLUSIONS: We demonstrate an evidence-based method to incorporate Na into MELD, which provides more accurate survival prediction than MELD alone. PMID- 16697730 TI - Liver microsomal triglyceride transfer protein is involved in hepatitis C liver steatosis. AB - BACKGROUND & AIMS: Hepatic steatosis is frequent in chronic hepatitis C. Several mechanisms might be implicated, including metabolic cofactors and direct viral effects on intracellular lipid pathways. In a transgenic mouse model, hepatitis C virus (HCV) was shown to inhibit microsomal triglyceride transfer protein (MTP) activity, which is essential for hepatic lipoprotein assembly and secretion. No data are available on liver MTP activity in HCV-infected patients. We therefore investigated liver MTP gene expression and its lipid transfer activity in untreated cases infected with the major HCV genotypes showing variable degrees of hepatic steatosis. METHODS: MTP messenger RNA (mRNA) levels were measured by real time polymerase chain reaction, and MTP activity was assessed by fluorescent assay in liver biopsy specimens of 58 HCV-positive patients. A set of metabolic and serum lipid markers was also measured at the time of liver biopsies. RESULTS: MTP mRNA levels showed a statistically significant (P = .001) inverse correlation with the degree of steatosis, independently of the HCV genotype. MTP mRNA levels also had an inverse correlation with serum insulin (P = .0002), homeostasis model assessment-insulin resistance (HOMA-IR) (P = .005), and body mass index (P = .02) in patients with HCV-1 and HCV-2 and with serum HCV-RNA (P = .02) in HCV-3 patients. Liver MTP-specific activity was significantly reduced in HCV-3 patients compared with those with other HCV genotypes (P = .004) and correlated with reduced serum cholesterol, apo B, and low-density lipoproteins. CONCLUSIONS: MTP may play a central role in HCV-related steatosis, being modulated by different genotype-specific mechanisms, mainly hyperinsulinemia in non-HCV-3 patients, and more profound and direct virus-related effects in HCV-3-infected individuals. PMID- 16697731 TI - Proteomic classification of pancreatic adenocarcinoma tissue using protein chip technology. AB - BACKGROUND & AIMS: Pancreatic adenocarcinoma is a most devastating cancer that presents late and is rapidly progressive. This study aimed to identify unique, tissue-specific protein biomarkers capable of differentiating pancreatic adenocarcinoma (PC) from adjacent uninvolved pancreatic tissue (AP), benign pancreatic disease (B), and nonmalignant tumor tissue (NM). METHODS: Tissue samples representing PC (n = 31), AP (n = 44), and B (n = 19) tissue were analyzed on hydrophobic protein chip arrays by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry. Training models were developed using logistic regression and validated using the 10-fold cross validation approach. RESULTS: The hydrophobic protein chip array revealed 13 protein peaks differentially expressed between PC and AP (receiver operating characteristic [ROC] area under the curve [AUC], 0.64-0.85), 8 between PC and B (ROC AUC, 0.67-0.78), and 12 between PC and NM tissue (ROC AUC, 0.63-0.81). Logistic regression and cross-validation identified overlapping panels of peaks to develop a training model that distinguished PC from AP (77.4% sensitivity, 84.1% specificity), B (83.9% sensitivity, 78.9% specificity), and NM tissue (58.1% sensitivity, 90.5% specificity). The final panels selected correctly classified 80.6% of PC and 88.6% of AP samples (ROC AUC, 0.92), 93.5% of PC and 89.5% of B samples (ROC AUC, 0.99), and 71.0% of PC and 92.1% of NM samples (ROC AUC, 0.91). CONCLUSIONS: This study used surface-enhanced laser desorption/ionization time-of-flight mass spectrometry to discover a number of protein panels that can distinguish effectively between pancreatic adenocarcinoma, benign, and adjacent pancreatic tissue. Identification of these proteins will add to our understanding of the biology of pancreatic cancer. Furthermore, these protein panels may have important diagnostic implications. PMID- 16697732 TI - Identification of two gene variants associated with risk of advanced fibrosis in patients with chronic hepatitis C. AB - BACKGROUND & AIMS: Previously identified clinical risk factors such as sex, alcohol consumption, and age at infection do not accurately predict which patients with chronic hepatitis C (CHC) will develop advanced fibrosis (bridging fibrosis and cirrhosis). The aim of this study was to identify genetic polymorphisms that can predict the risk of advanced fibrosis in patients with CHC. METHODS: A total of 916 subjects with CHC was enrolled from 2 centers. A gene-centric disease association study of 24,832 putative functional, single nucleotide polymorphisms (SNPs) was performed. Of the 1609 SNPs that were significantly associated (P 5% of its own weight of toxin in vitro. Administration of the GM(1)-expressing probiotic also protected infant mice against challenge with virulent V cholerae, even when treatment was delayed until after establishment of infection. When treatment commenced 1 hour after challenge, 12 of 12 mice given the probiotic survived, compared with only 1 of 12 for control mice (P < .00001). CONCLUSIONS: Toxin binding probiotics such as that described here have considerable potential for prophylaxis and treatment of cholera in humans. PMID- 16697734 TI - Trefoil factor family-1 mutations enhance gastric cancer cell invasion through distinct signaling pathways. AB - BACKGROUND & AIMS: Trefoil factor family-1 (TFF1) is a key gastric tumor suppressor gene. TFF1 knockout mice develop multiple gastric adenomas and carcinomas, and human gastric cancers typically lack TFF1 expression. Recently, TFF1 mutations have been found in human gastric cancer. The purpose of this study was to determine the functionality of these mutants. METHODS: Recombinant wild type TFF1 and the gastric cancer-associated TFF1 mutants (A10D and E13K) were produced and tested for their effect on gastric cancer cell proliferation, apoptosis, and invasion. Molecular modeling was used to guide the choice of mutants and to evaluate structure-function relationships. RESULTS: Molecular modeling suggested that A10D and E13K altered the surface charge of the loop 1 region of TFF1 without disturbing protein stability. Recombinant wild-type TFF1 significantly inhibited cell growth; A10D and E13K lost this tumor-suppressive property along with the ability to block etoposide-induced apoptosis. Although wild-type TFF1 promoted cell invasion, A10D and E13K were even more pro-invasive. Invasion induced by both mutants was blocked by inhibiting PI3-kinase or phospholipase-C, but inhibiting Rho-associated kinase (ROCK) blocked only E13K induced invasion. CONCLUSIONS: The loss of tumor-suppressor activity and gain of invasiveness from single point mutations constitute evidence for a functional role of TFF1 mutations in gastric cancer. These site-directed mutagenesis experiments provide the tools for continued probing of signal transduction mechanisms and structural elements responsible for TFF1 functions. PMID- 16697735 TI - Therapeutic action of ghrelin in a mouse model of colitis. AB - BACKGROUND & AIMS: Ghrelin is a novel growth hormone-releasing peptide with potential endogenous anti-inflammatory activities ameliorating some pathologic inflammatory conditions. Crohn's disease is a chronic debilitating disease characterized by severe T helper cell (Th)1-driven inflammation of the colon. The aim of this study was to investigate the therapeutic effect of ghrelin in a murine model of colitis. METHODS: We examined the anti-inflammatory action of ghrelin in the colitis induced by intracolonic administration of trinitrobenzene sulfonic acid. Diverse clinical signs of the disease were evaluated, including weight loss, diarrhea, colitis, and histopathology. We also investigated the mechanisms involved in the potential therapeutic effect of ghrelin, such as inflammatory cytokines and chemokines, Th1-type response, and regulatory factors. RESULTS: Ghrelin ameliorated significantly the clinical and histopathologic severity of the trinitrobenzene sulfonic acid-induced colitis; abrogating body weight loss, diarrhea, and inflammation; and increasing survival. The therapeutic effect was associated with down-regulation of both inflammatory and Th1-driven autoimmune response through the regulation of a wide spectrum of inflammatory mediators. In addition, a partial involvement of interluekin-10/transforming growth factor-beta1-secreting regulatory T cells in this therapeutic effect was demonstrated. Importantly, the ghrelin treatment was therapeutically effective in established colitis and avoided the recurrence of the disease. CONCLUSIONS: Our data demonstrate novel anti-inflammatory actions for ghrelin in the gastrointestinal tract, ie, the capacity to deactivate the intestinal inflammatory response and to restore mucosal immune tolerance at multiple levels. Consequently, ghrelin administration represents a novel possible therapeutic approach for the treatment of Crohn's disease and other Th1-mediated inflammatory diseases, such as rheumatoid arthritis and multiple sclerosis. PMID- 16697736 TI - CD4+ T-cell modulation of visceral nociception in mice. AB - BACKGROUND & AIMS: Although inflammatory and immune cells are present in the gut in the absence of pathology, their presence does not result in sensitization of sensory nerves, implying the existence of a local antinociceptive influence. We hypothesized that a component of the immune system exerts an antinociceptive influence, thus enabling the gut to function in the absence of undue pain or discomfort. METHODS: Visceromotor responses to colorectal distention were measured in mice with severe combined immune deficiency (SCID) and their wild type controls. RESULTS: SCID mice exhibited significantly lower pain thresholds. Transfer of CD4(+) T, but not B lymphocytes, normalized visceral pain in these mice. The restoration of normal visceral nociception following T-cell reconstitution in SCID mice was blocked by naloxone methiodide. Using an enzyme immunoassay and immunohistochemistry for beta-endorphin, we showed that in vitro stimulation of T lymphocytes induced the synthesis and release of beta-endorphin and that transfer of T cells into SCID mice increased the expression of beta endorphin in the enteric nervous system. CONCLUSIONS: These findings indicate that the immune system is a critical determinant of visceral nociception and that T lymphocytes provide an important opioid-mediated antinociceptive influence in the gut. PMID- 16697737 TI - The role of neurokinin 1 receptors in the maintenance of visceral hyperalgesia induced by repeated stress in rats. AB - BACKGROUND & AIMS: The neurokinin 1 receptors (NK(1)Rs) and substance P (SP) have been implicated in the stress and/or pain pathways involved in chronic pain conditions. Here we examined the participation of NK(1)Rs in sustained visceral hyperalgesia observed in rats exposed to chronic psychological stress. METHODS: Male Wistar rats were exposed to daily 1-hour water avoidance stress (WA) or sham WA for 10 consecutive days. We tested intraperitoneal or intrathecal injection of the NK(1)R antagonist SR140333 on the visceromotor reflex to colorectal distention in both groups at day 11. Real-time reverse-transcription polymerase chain reaction, Western blot, and immunohistochemistry were used to assess the expression of NK(1)Rs and/or SP in samples of colon, spinal cord, and dorsal root ganglia. RESULTS: Both intraperitoneal and intrathecal SR140333 injection diminished the enhanced visceromotor reflex to colorectal distention at day 11 in stressed rats but did not affect the response in control animals. Real-time polymerase chain reaction and Western blotting demonstrated stress-induced up regulation of spinal NK(1)Rs. Immunohistochemistry showed an increased number of NK(1)R-expressing neurons in the laminae I of the dorsal horn in stressed rats. The expression of NK(1)Rs was decreased in colon from stressed rats compared with control. The expression of SP gene precursor in dorsal root ganglia was unchanged in stressed rats compared with controls. CONCLUSIONS: Stress-induced increased NK(1)R expression on spinal neurons and the inhibitory effect of intrathecal NK(1)R antagonist on visceral hyperalgesia support the key contribution of spinal NK(1)Rs in the molecular pathways involved in the maintenance of visceral hyperalgesia observed after chronic WA. PMID- 16697738 TI - Antidepressants attenuate increased susceptibility to colitis in a murine model of depression. AB - BACKGROUND & AIMS: Psychiatric factors may determine gastrointestinal health outcomes. Here, we used a model of depression based on neonatal maternal separation (MS) to identify alterations in gut physiology and to assess its association with increased sensitivity to experimental colitis in adulthood. We also examined whether antidepressant therapy attenuates the increased susceptibility to colitis. METHODS: C57BL/6 mouse pups were separated from mothers for 3 hours per day at 1-21 days of age. Maternally unseparated (US) litters served as controls. At 8 weeks of age mice were examined for changes in behavior, intestinal permeability, and sensitivity to colitis. Separate sets of MS and US mice were given either saline or the antidepressant desipramine 15 mg/kg once daily at 23-36 days of age. Testing of mice occurred at 8 weeks of age. RESULTS: Adult MS mice showed evidence of depressive-like behavior and enhanced intestinal permeability but showed no evidence of spontaneous inflammation. A more severe colitis was seen in MS compared with US mice. Antidepressant therapy improved parameters of depressive-like behavior and reduced the vulnerability to dextran sulphate sodium colitis in MS mice but had no effect on colitis in US mice. CONCLUSIONS: MS may lead to depression and increased responsiveness to stress, to impaired intestinal barrier function, and to enhanced vulnerability to colitis in adulthood. This vulnerability is reversed by antidepressant therapy. Depression increases vulnerability to intestinal inflammation. We speculate that pre-existing depression may facilitate the expression of inflammatory bowel diseases. PMID- 16697739 TI - The role of matrix metalloproteinase-7 in redefining the gastric microenvironment in response to Helicobacter pylori. AB - BACKGROUND & AIMS: Interactions between epithelial and stromal cells are important determinants of mucosal organization, but the signaling mechanisms are understood incompletely. Matrix metalloproteinase (MMP)-7 is produced uniquely in epithelia, may act on growth factors and matrix proteins, and in the stomach is increased with Helicobacter pylori infection. We have studied the role of MMP-7 in signaling between epithelial cells and a key stromal cell type, the myofibroblast. METHODS: Immunohistochemistry and Western blotting were applied to gastric corpus biopsy specimens; primary cultures of human gastric glands and myofibroblasts were used to study the role of MMP-7 in regulating proliferation and migration of the latter, and MMP-7 substrates were identified by proteomic methods. RESULTS: Increased abundance of the myofibroblast marker alpha-smooth muscle actin was identified in H. pylori-positive biopsy specimens. Media from H pylori-infected gastric epithelial cultures stimulated proliferation and migration of primary human gastric myofibroblasts and antisense oligonucleotide treatment indicated a role for MMP-7. Proteomic methods identified insulin-like growth factor binding protein (IGFBP)-5 as a substrate for MMP-7 in medium from gastric myofibroblasts. Knockdown of IGFBP-5 by small interfering RNA or immunoneutralization of IGF-II, abolished myofibroblast responses to MMP-7. Proliferation of gastric epithelial cells also was stimulated by MMP-7-treated myofibroblasts via IGF-II. CONCLUSIONS: MMP-7 acts as an epithelial-derived signal increasing the bioavailability of IGF-II released from myofibroblasts. Because IGF-II acts on both stromal and epithelial cells, the findings suggest that increased MMP-7 expression contributes to redefining the niche occupied by dividing cells and leading to hyperproliferation in H pylori infection. PMID- 16697740 TI - Candida albicans is an immunogen for anti-Saccharomyces cerevisiae antibody markers of Crohn's disease. AB - BACKGROUND AND AIMS: Antibodies directed against oligomannose sequences alpha-1,3 Man (alpha-1,2 Man alpha-1,2 Man)(n) (n = 1 or 2), termed anti-Saccharomyces cerevisiae antibodies (ASCAs) are markers of Crohn's disease (CD). S. cerevisiae mannan, which expresses these haptens, is used to detect ASCA, but the exact immunogen for ASCA is unknown. Structural and genetic studies have shown that Candida albicans produces mannosyltransferase enzymes that can synthesize S cerevisiae oligomannose sequences depending on growth conditions. This study investigated whether C. albicans could act as an immunogen for ASCA. METHODS: Sequential sera were collected from patients with CD, systemic candidiasis, and rabbits infected with C. albicans. Antibodies were purified by using chemically synthesized (Sigma) ASCA major epitopes. These affinity-purified antibodies and lectins were then used to analyze the expression of ASCA epitopes on molecular extracts and cell walls of C. albicans and S cerevisiae grown in various conditions. RESULTS: In humans and rabbits, generation of ASCA was shown to be associated with the generation of anti-C. albicans antibodies resulting specifically from infection. By using affinity-purified antibodies, C. albicans was shown to express ASCA epitopes on mannoproteins similar to those of S. cerevisiae. By changing the growth conditions, C. albicans mannan was also able to mimic S. cerevisiae mannan in its ability to detect ASCA associated with CD. This overexpression of ASCA epitopes was achieved when C. albicans grew in human tissues. CONCLUSIONS: C. albicans is one of several immunogens for ASCA and may be at the origin of an aberrant immune response in CD. PMID- 16697741 TI - Diet- and colonization-dependent intestinal dysfunction predisposes to necrotizing enterocolitis in preterm pigs. AB - BACKGROUND & AIMS: Preterm birth and formula feeding are key risk factors associated with necrotizing enterocolitis (NEC) in infants, but little is known about intestinal conditions that predispose to disease. Thus, structural, functional, and microbiologic indices were used to investigate the etiology of spontaneous NEC development in preterm pigs. METHODS: Piglets were delivered by cesarean section at 92% gestation, reared in infant incubators, and fed infant formula or colostrum every 3 hours (n = 120) until tissue collection at 1-2 days of age. RESULTS: Clinical and histopathologic signs of NEC were observed in 57% of pigs fed FORMULA (26/46) and in 5% of pigs fed COLOSTRUM (2/38) (P < .05). Relative to COLOSTRUM, both healthy and sick FORMULA pigs had reduced intestinal villous heights, enzyme activities, nutrient absorption, and antioxidant levels and higher inducible nitric oxide synthetase activity (P < .05). In healthy pigs, mucosal microbial diversity remained low and diet independent. NEC pigs showed bacterial overgrowth, and a high mucosal density of Clostridium perfringens was detected in some but not all pigs. Germ-free conditions and antiserum against Clostridium perfringens toxin prevented intestinal dysfunction and NEC in formula fed pigs, whereas the gut trophic factors, epidermal growth factor, and glucagon like peptide 2 had limited effects. CONCLUSIONS: A subclinical, formula-induced mucosal atrophy and dysfunction predispose to NEC and bacterial overgrowth. The adverse feeding effects are colonization dependent and may be reduced by factors in colostrum that include antibodies against aggressive toxins such as those of Clostridium perfringens. PMID- 16697742 TI - Drug and bile acid transporters in rosuvastatin hepatic uptake: function, expression, and pharmacogenetics. AB - BACKGROUND & AIMS: The 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, or statins, target liver HMG-CoA and are of proven benefit in the prevention of coronary heart disease. Rosuvastatin is an effective statin notable for liver selectivity and lack of significant metabolism. We assessed the extent and relevance of hepatic transporters to rosuvastatin uptake. METHODS: Transporters involved in rosuvastatin uptake were determined through heterologous expression of multiple human and rat uptake transporters. Human organic anion transporting polypeptide (OATP) 1B1 and sodium-dependent taurocholate cotransporting polypeptide (NTCP) allelic variants were also assessed. Expression of OATP and NTCP messenger RNA and protein was determined from a bank of human liver samples. RESULTS: Multiple OATP family members, including 1B1, 1B3, 2B1, and 1A2, were capable of rosuvastatin transport. Naturally occurring polymorphisms in OATP1B1, including *5, *9, *15, and *18, were associated with profound loss of activity toward rosuvastatin. Interestingly, the major human hepatic bile acid uptake transporter NTCP, but not rat Ntcp, also transported rosuvastatin. Human hepatocyte studies suggested that NTCP alone accounted for approximately 35% of rosuvastatin uptake. Remarkably, NTCP*2, a variant known to have a near complete loss of function for bile acids, exhibited a profound gain of function for rosuvastatin. Quantitative messenger RNA analysis revealed marked intersubject variability in expression of OATPs and NTCP. CONCLUSIONS: Multiple transporters mediate the overall hepatic uptake of rosuvastatin, and NTCP may be a heretofore unrecognized transporter important to the disposition of rosuvastatin and possibly other drugs/statins in clinical use. Accordingly, transporter expression and polymorphisms may be key determinants of intersubject variability in response to statin therapy in general. PMID- 16697743 TI - The bone marrow functionally contributes to liver fibrosis. AB - BACKGROUND & AIMS: Bone marrow (BM) cells may transdifferentiate into or fuse with organ parenchymal cells. BM therapy shows promise in murine models of cirrhosis, and clinical trials of bone marrow stem cell therapy for organ healing are underway. However, the BM may contribute to scar-forming myofibroblasts in various organs including the liver. We have studied this axis of regeneration and scarring in murine models of cirrhosis, including an assessment of the temporal and functional contribution of the BM-derived myofibroblasts. METHODS: Female mice were lethally irradiated and received male BM transplants. Carbon tetrachloride or thioacetamide was used to induce cirrhosis. BM-derived cells were tracked through in situ hybridization for the Y chromosome. BM transplants from 2 strains of transgenic mice were used to detect intrahepatic collagen production. RESULTS: In the cirrhotic liver, the contribution of BM to parenchymal regeneration was minor (0.6%); by contrast, the BM contributed significantly to hepatic stellate cell (68%) and myofibroblast (70%) populations. These BM-derived cells were found to be active for collagen type 1 transcription in 2 independent assays and could influence the fibrotic response to organ injury. These BM-derived myofibroblasts did not occur through cell fusion between BM-derived cells and indigenous hepatic cells but, instead, originated largely from the BM's mesenchymal stem cells. CONCLUSIONS: The BM contributes functionally and significantly to liver fibrosis and is a potential therapeutic target in liver fibrosis. Clinical trials of BM cell therapy for liver regeneration should be vigilant for the possibility of enhanced organ fibrosis. PMID- 16697744 TI - The vagus nerve and nicotinic receptors modulate experimental pancreatitis severity in mice. AB - BACKGROUND & AIMS: The nervous system, through the vagus nerve, controls inflammation by decreasing the release of tumor necrosis factor-alpha from endotoxin stimulated macrophages. This anti-inflammatory effect is mediated by an interaction of acetylcholine, the principal neurotransmitter of the vagus nerve, with macrophage cholinergic nicotinic receptors expressing the alpha7 subunit. METHODS: To determine the role of this "nicotinic anti-inflammatory pathway" in experimental pancreatitis, we induced pancreatitis in mice by 12 hourly intraperitoneal injections of cerulein. Pancreatitis was preceded by unilateral left cervical vagotomy or pretreatment with the nicotinic receptor antagonist mecamylamine or by pretreatment with the selective alpha7 nicotinic receptor agonist 3-(2,4-dimethoxybenzylidene) anabaseine (GTS-21). RESULTS: Vagotomy or pretreatment with mecamylamine resulted in an enhanced severity of pancreatitis, as reflected by histology, edema, plasma hydrolases, and interleukin-6 levels. Furthermore, the number of neutrophils migrated to the pancreas was increased in these mice, as shown by myeloperoxidase content and intrapancreatic staining of neutrophils. Conversely, GTS-21 pretreatment strongly decreased the severity of pancreatitis. Pancreatitis-associated pulmonary inflammation was independent of the integrity of the vagus nerve and nicotinic receptors. CONCLUSIONS: This study provides the first evidence for a therapeutic potential of the vagus nerve and the "nicotinic anti-inflammatory pathway" in attenuating inflammation and injury during experimental pancreatitis. PMID- 16697745 TI - Endogenous opioids modulate the growth of the biliary tree in the course of cholestasis. AB - BACKGROUND & AIMS: There is poor knowledge on the factors that modulate the growth of cholangiocytes, the epithelial cell target of cholangiopathies, which are diseases leading to progressive loss of bile ducts and liver failure. Endogenous opioids are known to modulate cell growth. In the course of cholestasis, the opioidergic system is hyperactive, and in cholangiocytes a higher expression of opioid peptide messenger RNA has been described. This study aimed to verify if such events affect the cholangiocyte proliferative response to cholestasis. METHODS: The presence of the delta opioid receptor (OR), muOR, and kappaOR was evaluated. The effects on cholangiocyte proliferation of the in vitro and in vivo exposure to their selective agonists, together with the intracellular signals, were then studied. The effects of the OR antagonist naloxone on cell growth were also tested both in vivo and in vitro. RESULTS: Cholangiocytes express all 3 receptors studied. deltaOR activation strongly diminished the proliferative and functional response of cholangiocytes to cholestasis, whereas muOR resulted in a slight increase in cell growth. The deltaOR signal is mediated by the IP3/CamKIIalpha/PKCalpha pathway, which inhibits the cAMP/PKA/ERK1/2/AKT cascade. In contrast, muOR activation stimulates the cAMP/PKA/ERK1/2/AKT cascade but does not affect the IP3/CamKIIalpha/PKCalpha pathway. The blockage of endogenous opioid peptides by naloxone further enhanced cholangiocyte growth both in vivo and in vitro. CONCLUSIONS: The increase in opioid peptide synthesis in the course of cholestasis aims to limit the excessive growth of the biliary tree in the course of cholestasis by the interaction with the deltaOR expressed by cholangiocytes. PMID- 16697746 TI - NAFLD, obesity, and bariatric surgery. PMID- 16697747 TI - Intestinal cholesterol absorption inhibitor ezetimibe added to cholestyramine for sitosterolemia and xanthomatosis. AB - Sitosterolemia is a rare, recessively inherited disorder characterized by increased absorption and delayed removal of noncholesterol sterols, which is associated with accelerated atherosclerosis, premature coronary artery disease, hemolysis, and xanthomatosis. Treatments include low-sterol diet and bile salt binding resins; however, these often do not reduce the xanthomatosis. We examined the effects of the intestinal cholesterol/phytosterol transporter inhibitor ezetimibe added to cholestyramine in a young female patient with sitosterolemia and associated xanthomatosis. The patient was an 11-year-old female with sitosterolemia presenting with prominent xanthomas in the subcutaneous tissue of both elbows who was receiving treatment with cholestyramine 2 g once daily. Bilateral carotid bruits were audible, and a grade II/VI systolic murmur was detected at the left upper sternal border. She also had a low platelet count of 111,000/microL. Ezetimibe 10 mg once daily was added to the patient's ongoing cholestyramine regimen, and she was evaluated for 1 year. The patient followed an unrestricted diet during the 1-year treatment period. After 1 year of treatment with ezetimibe added to ongoing cholestyramine therapy, the patient's plasma sitosterol and campesterol levels decreased by approximately 50%. Her carotid bruits completely resolved, her systolic murmur diminished, and her platelet count rose to 268,000/microL. More remarkably, the tuberous xanthomas on her elbows had completely regressed. Ezetimibe added to ongoing low-dose cholestyramine therapy led to a marked improvement in plasma sterol concentrations, complete regression of xanthomatosis, resolution of carotid bruits, and improvement in cardiac murmur in a young female patient with sitosterolemia. PMID- 16697748 TI - Diagnosis of gastric cancer by serum proteomic fingerprinting. AB - BACKGROUND & AIMS: Accurate serum biomarkers for gastric cancer currently are lacking. We attempted to identify potential diagnostic serum markers for gastric cancer with the use of the surface-enhanced laser desorption/ionization ProteinChip technology. METHODS: The study was divided into 3 phases: (1) discovery of potential diagnostic markers using sera of gastric cancer patients and controls, (2) development of a diagnostic model, and (3) independent validation of the diagnostic model using a different cohort of gastric cancer and control patients. The serum proteins/peptides were analyzed with 2 types of ProteinChip arrays, IMAC30 arrays loaded with copper (II) ion and CM10 (weak cation exchange) arrays. RESULTS: In the discovery set, peak intensities of 31 surface-enhanced laser desorption/ionization proteomic features were significantly higher in gastric cancer patients. The tumor-specific nature of 6 proteomic features with the mass/charge (m/z) values of 5098, 8592, 8610, 11,468, 11,804, and 50,140 was verified by their lower peak intensities in postoperative sera. After excluding the sodium adduct peak (8610 m/z) of the 8592 m/z protein, the peak intensities of the tumor-specific proteomic features were used to develop a linear regression model for calculating a diagnostic index. The area under the receiver operating characteristic curve of the corresponding diagnostic index was 0.92 (95% confidence interval, 0.85-0.99) in the independent validation set. At a specificity of 95%, the sensitivity for gastric cancer detection was 83%. CONCLUSIONS: A unique serum proteomic fingerprint can be detected in the sera of gastric cancer patients, which may be useful in the noninvasive diagnosis of gastric cancer. PMID- 16697749 TI - Guidelines for colonoscopy surveillance after cancer resection: a consensus update by the American Cancer Society and the US Multi-Society Task Force on Colorectal Cancer. AB - Patients with resected colorectal cancer are at risk for recurrent cancer and metachronous neoplasms in the colon. This joint update of guidelines by the American Cancer Society and the US Multi-Society Task Force on Colorectal Cancer addresses only the use of endoscopy in the surveillance of these patients. Patients with endoscopically resected Stage I colorectal cancer, surgically resected Stages II and III cancers, and Stage IV cancer resected for cure (isolated hepatic or pulmonary metastasis) are candidates for endoscopic surveillance. The colorectum should be carefully cleared of synchronous neoplasia in the perioperative period. In nonobstructed colons, colonoscopy should be performed preoperatively. In obstructed colons, double-contrast barium enema or computed tomography colonography should be performed preoperatively, and colonoscopy should be performed 3 to 6 months after surgery. These steps complete the process of clearing synchronous disease. After clearing for synchronous disease, another colonoscopy should be performed in 1 year to look for metachronous lesions. This recommendation is based on reports of a high incidence of apparently metachronous second cancers in the first 2 years after resection. If the examination at 1 year is normal, then the interval before the next subsequent examination should be 3 years. If that examination is normal, then the interval before the next subsequent examination should be 5 years. Shorter intervals may be indicated by associated adenoma findings (see "Guidelines for Colonoscopy Surveillance After Polypectomy: A Consensus Update by the US Multi Society Task Force on Colorectal Cancer and the American Cancer Society"). Shorter intervals also are indicated if the patient's age, family history, or tumor testing indicate definite or probable hereditary nonpolyposis colorectal cancer. Patients undergoing low anterior resection of rectal cancer generally have higher rates of local cancer recurrence compared with those with colon cancer. Although effectiveness is not proven, performance of endoscopic ultrasound or flexible sigmoidoscopy at 3- to 6-month intervals for the first 2 years after resection can be considered for the purpose of detecting a surgically curable recurrence of the original rectal cancer. PMID- 16697750 TI - Guidelines for colonoscopy surveillance after polypectomy: a consensus update by the US Multi-Society Task Force on Colorectal Cancer and the American Cancer Society. AB - Adenomatous polyps are the most common neoplastic findings discovered in people who undergo colorectal screening or who have a diagnostic work-up for symptoms. It was common practice in the 1970s for these patients to have annual follow-up surveillance examinations to detect additional new adenomas and missed synchronous adenomas. As a result of the National Polyp Study report in 1993, which showed clearly in a randomized design that the first postpolypectomy examination could be deferred for 3 years, guidelines published by a gastrointestinal consortium in 1997 recommended that the first follow-up surveillance take place 3 years after polypectomy for most patients. In 2003 these guidelines were updated and colonoscopy was recommended as the only follow up examination, stratification at baseline into low risk and higher risk for subsequent adenomas was suggested. The 1997 and 2003 guidelines dealt with both screening and surveillance. However, it has become increasingly clear that postpolypectomy surveillance is now a large part of endoscopic practice, draining resources from screening and diagnosis. In addition, surveys have shown that a large proportion of endoscopists are conducting surveillance examinations at shorter intervals than recommended in the guidelines. In the present report, a careful analytic approach was designed to address all evidence available in the literature to delineate predictors of advanced pathology, both cancer and advanced adenomas, so that patients can be stratified more definitely at their baseline colonoscopy into those at lower risk or increased risk for a subsequent advanced neoplasia. People at increased risk have either 3 or more adenomas, high grade dysplasia, villous features, or an adenoma 1 cm or larger in size. It is recommended that they have a 3-year follow-up colonoscopy. People at lower risk who have 1 or 2 small (<1 cm) tubular adenomas with no high-grade dysplasia can have a follow-up evaluation in 5-10 years, whereas people with hyperplastic polyps only should have a 10-year follow-up evaluation, as for average-risk people. There have been recent studies that have reported a significant number of missed cancers by colonoscopy. However, high-quality baseline colonoscopy with excellent patient preparation and adequate withdrawal time should minimize this and reduce clinicians concerns. These guidelines were developed jointly by the US Multi-Society Task Force on Colorectal Cancer and the American Cancer Society to provide a broader consensus and thereby increase the use of the recommendations by endoscopists. The adoption of these guidelines nationally can have a dramatic impact on shifting available resources from intensive surveillance to screening. It has been shown that the first screening colonoscopy and polypectomy produces the greatest effects on reducing the incidence of colorectal cancer in patients with adenomatous polyps. PMID- 16697751 TI - Toll-like receptor signaling in the liver. PMID- 16697752 TI - Our new president--Mark Donowitz, MD. PMID- 16697753 TI - Alcohol and hepatitis C: do we have a drinking problem? PMID- 16697754 TI - Genetically engineered probiotic competition. PMID- 16697755 TI - Cancer serum proteomics in gastroenterology. PMID- 16697756 TI - Child, MELD, hyponatremia, and now portal pressure. PMID- 16697758 TI - A hair of the dog that bit you. PMID- 16697759 TI - MLN-02 in IBD: is "super-selective" really super? PMID- 16697760 TI - The skinny on obesity and reflux. PMID- 16697761 TI - CLASSIC-I study the efficacy of adalimumab. PMID- 16697762 TI - Adalimumab induction for Crohn's disease. PMID- 16697763 TI - Is there a role for sequential in sequential anti-H. pylori therapy? PMID- 16697765 TI - Coffee and tea consumption and chronic liver disease. PMID- 16697768 TI - Amadori-modified glycated serum proteins and accelerated atherosclerosis in diabetes: pathogenic and therapeutic implications. PMID- 16697769 TI - Aspirin therapy for inhibition of platelet reactivity in the presence of erythrocytes in patients with vascular disease. AB - Inhibition of erythrocyte (RBC) promotion of platelet reactivity could improve the antiplatelet effect of aspirin (ASA). We tested different ASA regimens for optimal inhibition of platelets and the effects of RBC in patients with a history of vascular diseases. Collagen-induced platelet activation (14C-5HT, TXA2 release) and platelet recruitment (proaggregatory activity of cell-free releasates from activated platelets) were measured in PRP, platelet-RBC (Hct 40%), and whole blood (WB) in 206 patients initially on 200-300-mg ASA/day. Their regimen was modified to biweekly 500 mg (loading dose, L) plus daily or twice daily low-dose ASA (50 or 100 mg). TXA2 was inhibited with all regimens. Percentage of patients with suboptimal inhibition of platelet recruitment in WB was 200-300 ASA/day (41%), L-50/day (87%), L-100/day (58%), L-50/twice-daily (39%), and L-100/twice-daily (20%; P < 0.05 vs other regimens). 14C-5HT release was inhibited to the greatest extent with L-100/twice-daily in PRP + RBC or WB (P < 0.05 vs other regimens) due to greater inhibition of the RBC prothrombotic effect. Compared with other ASA regimens, L-100 twice-daily (equivalent to 221-mg ASA/day in the 14-day cycle), reduced by >50% the proportion of patients with suboptimal inhibition of platelet recruitment in WB and inhibited 14C-5HT release to the greatest extent. PMID- 16697770 TI - Polymorphisms of p53 and p21 genes in chronic obstructive pulmonary disease. AB - BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a multifactorial disease influenced by genetic and environmental factors, particularly cigarette smoking. Although cigarette smoke may be directly mutagenic, polymorphisms in the genes controlling acquired somatic mutations may also contribute, at least to some extent, to the observed differing susceptibilities to COPD. To investigate the involvement of genetic polymorphisms of p53 and p21 in the pathogenesis of COPD, the authors performed a case-control study involving 206 subjects with COPD and 210 healthy smokers as control subjects. METHODS: Polymorphisms of p53 and p21 genes were analyzed using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique on genomic DNA isolated from peripheral lymphocytes. The distribution of the p53 and p21 polymorphisms in healthy subjects and COPD patients was examined and compared using the Pearson X2 test. Significance was accepted at P < 0.05. Odds ratios (ORs) and 95% confidence intervals (CIs) of each specific genotype were calculated using logistic regression analysis to quantitatively assess the degree of association observed. RESULTS: The distribution frequencies of genotypes of p53 codon 72 and p21 codon 31 were significantly different between the COPD and the control groups. Higher ORs for COPD were seen for persons with p53 Pro/Pro or Pro/Arg genotypes against Arg/Arg genotype [OR = 2.35, 95% CI 1.27-4.39, P = 0.008], or p21 Arg/Arg and Arg/Ser genotypes against Ser/Ser genotype [OR = 2.07, 95% CI 1.06-4.05, P = 0.033]. CONCLUSIONS: The polymorphisms of p53 and p21 were significantly associated with the occurrence of smoking-related COPD in Taiwan Chinese patients. PMID- 16697771 TI - Coordinate activation of intracellular signaling pathways by insulin-like growth factor-1 and platelet-derived growth factor in rat hepatic stellate cells. AB - Proliferation of activated hepatic stellate cells (HSC) is an important event in the development of hepatic fibrosis. Insulin-like growth factor-1 (IGF-1) has been shown to be mitogenic for HSC, but the intracellular signaling pathways involved have not been fully characterized. Thus, the aims of the current study were to examine the roles of the extracellular signal-regulated kinase (ERK), phosphatidylinositol 3-kinase (PI3-K) and p70-S6 kinase (p70-S6-K) signaling pathways in IGF-1- and platelet-derived growth factor (PDGF)-induced mitogenic signaling of HSC and to examine the potential crosstalk between these pathways. Both IGF-1 and PDGF increased ERK, PI3-K and p70-S6-K activity. When evaluating potential crosstalk between these signaling pathways, we observed that PI3-K is required for p70-S6-K activation by IGF-1 and PDGF, and is partially responsible for PDGF-induced ERK activation. PDGF and IGF-1 also increased the levels of cyclin D1 and phospho-glycogen synthase kinase-3beta. Coordinate activation of ERK, PI3-K and p70-S6-K is important for perpetuating the activated state of HSC during fibrogenesis. PMID- 16697772 TI - Characterization of peripheral natural killer cells in primary Sjogren's syndrome: impaired NK cell activity and low NK cell number. AB - The aim of this study was to compare the number of peripheral blood natural killer (NK) cells, NK cell activity, expression of NK cell activating receptors, and serum cytokine levels in patients with primary Sjogren's syndrome (SS) vs normal controls. The authors found that NK cell number, NK cell killing activity, and the expression of activating receptors CD2 and NKG2D were significantly decreased, and the expression of NKp46, as well as the percentage of apoptotic NK cells, were significantly increased in primary SS patients compared with healthy controls. NK cell killing activity on a per-cell basis was similar in primary SS patients and healthy controls. Moreover, the levels of IL-18 and TNF-alpha, cytokines that have been shown to promote NK cell death, were significantly increased in sera from patients with primary SS compared with controls. These data suggest that reduced NK cell numbers, probably a result of apoptotic death, may contribute to impaired NK cell activity in patients with primary SS. PMID- 16697773 TI - Differential gene expression identifies subgroups of renal cell carcinoma. AB - Clear cell carcinoma of the kidney, the most common subtype of renal cell cancer, displays different biological behavior in different patients. This heterogeneity cannot be recognized by light microscopy. In this study, gene expression in 16 clear cell renal cell carcinoma samples and 17 non-malignant tissue types comprising 539 samples was determined using oligonucleotide microarrays representing approximately 40,000 known genes and ESTs. Differences in gene expression were quantified as the fold change in gene expression between the various sets of samples. A set of genes was identified that was overexpressed in the renal cell carcinoma samples compared with the normal kidney samples. Principle component analysis of the set of renal cell carcinomas using this set of genes overexpressed in renal cell cancer revealed the existence of 2 major subgroups among the renal carcinomas. A series of principle component analyses of the set of renal cell carcinomas using different gene sets composed of genes involved in different metabolic pathways also revealed the same 2 major subgroups of the renal cell cancers. Eisen clustering using the same genes also revealed the same 2 major renal cell cancer subsets. Review of the pathology suggested that these 2 subgroups differed in pathologic grade. Genes differentially expressed between the 2 renal cell cancer subsets were identified. Examination of gene expression in each renal cell cancer subset and the pool of renal cell carcinoma samples compared with that in 17 different normal tissues revealed genes specifically overexpressed in renal cell cancer compared with these normal tissues. The authors conclude that gene expression patterns may be useful in helping to further classify subtypes of renal cell carcinoma that may have clinical significance. In addition, the genes identified as overexpressed in each set of clear cell renal cell carcinomas compared with normal tissues may represent useful targets for therapy. PMID- 16697779 TI - The future of journals and meetings--is there one? PMID- 16697780 TI - Targeting the genetic basis of transitional cell carcinoma. PMID- 16697781 TI - Bladder cancer: in search of the right operation for the right patient. PMID- 16697782 TI - A new classification is needed for pelvic pain syndromes--are existing terminologies of spurious diagnostic authority bad for patients? PMID- 16697783 TI - Stimulating aspects of sacral nerve stimulation. PMID- 16697784 TI - The evolution of laparoscopy in pediatric urology--intelligent design? PMID- 16697785 TI - Papillary urothelial neoplasm of low malignant potential: evolving terminology and concepts. AB - PURPOSE: The most controversial aspect of the new WHO 2004/ISUP classification system is the creation of the PUNLMP diagnostic category. We discuss PUNLMP tumors and the WHO 2004/ISUP classification system with an emphasis on tumor morphology and heterogeneity, recurrence and progression rates, tumor genetics, interobserver variability and the usefulness of biomarkers and molecular diagnostic techniques for grading bladder tumors. MATERIALS AND METHODS: A literature search using PubMed was performed. All relevant literature concerning PUNLMP and the WHO 2004/ISUP grading system for urothelial neoplasms was reviewed. RESULTS: The new WHO 2004/ISUP classification reflects work in progress. Low malignant potential terminology may not reflect the true biological behavior of these tumors. Additionally, interobserver variability in making a diagnosis of PUNLMP is high despite detailed histological criteria. Urine cytopathology in the context of the WHO 2004/ISUP classification does not appear to effectively discriminate PUNLMP from low grade carcinoma. CONCLUSIONS: For practical purposes patients with PUNLMP should be treated similarly to patients with low grade, noninvasive urothelial carcinoma. It is our hope that recent advances in the molecular grading of these tumors may eventually supplant traditional morphological classification, allowing a more precise and objective assessment of the biological potential of these tumors. PMID- 16697786 TI - Complications of intravesical therapy for urothelial cancer of the bladder. AB - PURPOSE: Intravesical therapy is an integral part of treatment in patients with superficial urothelial carcinoma of the bladder. The American Urological Association and European Association of Urology guidelines on bladder cancer incorporate it for the treatment of superficial bladder cancer. Given the extensive use of intravesical immunotherapy and chemotherapy, it is essential for the practicing urologist to be aware of the local and systemic side effects of these therapies. MATERIALS AND METHODS: We reviewed the literature on intravesical immunotherapy and chemotherapy with particular emphasis on side effects, complications and their management. A Medline search of the English language literature for the last 25 years was done on Entrez PubMed and all relevant articles were studied in full. All side effects and complications were studied and their management was reviewed. RESULTS: Intravesical therapy for transitional cell carcinoma of the bladder is generally safe. There is a high incidence of local, usually self-limiting, relatively minor side effects and infrequent, potentially severe local and systemic side effects. Most side effects are avoidable. CONCLUSIONS: Although intravesical therapy is generally safe, local and systemic side effects occur and it is important to be aware of them. Identifying complications early, preventing them when possible and managing them efficiently are critical. Most complications are preventable. Knowing the risks and benefits of chemotherapeutic and immunotherapeutic agents may decrease the short-term and long-term toxicity of these agents. Adherence to guidelines may prevent inappropriate use, which can lead to unnecessary complications, resulting in bladder dysfunction and even cystectomy. PMID- 16697787 TI - Laparoscopic radical prostatectomy for localized prostate cancer: a systematic review of comparative studies. AB - PURPOSE: We compared the safety and efficacy of laparoscopic and open radical prostatectomy through a systematic assessment of the literature. MATERIALS AND METHODS: Literature databases were searched from 1996 to December 2004 inclusive. Studies comparing transperitoneal laparoscopic radical prostatectomy, extraperitoneal endoscopic radical prostatectomy or robot assisted radical prostatectomy with open radical retropubic prostatectomy or radical perineal prostatectomy for localized prostate cancer were included. Comparisons between different laparoscopic approaches were also included. RESULTS: We identified 30 comparative studies, of which none were randomized controlled trials. There were 21 studies comparing laparoscopic with open prostatectomy with a total of 2,301 and 1,757 patients, respectively, and 9 comparing different laparoscopic approaches with a total of 1,148 patients. In terms of safety there did not appear to be any important differences in the complication rate between laparoscopic and open approaches. However, blood loss and transfusions were lower for laparoscopic approaches. In terms of efficacy operative time was longer for laparoscopic than for open prostatectomy but length of stay and duration of catheterization were shorter. Positive margin rates and recurrence-free survival were similar. Continence and potency were not well reported but they appeared similar for the 2 approaches. There were no important differences between laparoscopic approaches. CONCLUSIONS: Laparoscopic radical prostatectomy is emerging as an alternative to open radical prostatectomy but randomized, controlled trials considering patient relevant outcomes, such as survival, continence and potency, with sufficient followup are required to determine relative safety and efficacy. PMID- 16697788 TI - Renal cell carcinoma and end stage renal disease. AB - PURPOSE: Patients with ESRD secondary to acquired renal cystic disease have been reported to have a higher incidence of RCC than the general population. We examined the clinical and pathological significance of incidental renal masses in patients with ESRD. MATERIALS AND METHODS: From January 1994 to July 2000, 852 consecutive patients with ESRD who were being considered for renal transplantation at University of Mississippi Medical Center were evaluated with renal ultrasound as part of assessment for possible kidney transplantation. Those patients with ultrasound suspicious for a malignant renal lesion were further evaluated with CT of the abdomen with and without intravenous contrast medium. Any patient with CT findings suspicious for RCC was recommended to undergo radical nephrectomy before kidney transplantation. RESULTS: A total of 19 patients had CT criteria for a possible malignant renal lesion. Seven patients had Bosniak class 3 renal cysts and 12 patients had solid, enhancing renal masses. Of the patients 17 underwent radical nephrectomy. On pathological examination 14 patients had RCC with a 1.64% prevalence in the population screened. Mean Fuhrman nuclear grade in our patients was 2.45. CONCLUSIONS: RCC in patients with ESRD are of clinical significance, considering the size, grade, histology and pathological stage of these tumors. The higher prevalence of clinically significant RCC in patients with ESRD as well as the risk of cancer progression while patients are on immunosuppressive medications justifies screening for RCC in patients with ESRD who are awaiting renal transplantation. PMID- 16697790 TI - Simple enucleation for the treatment of renal cell carcinoma between 4 and 7 cm in greatest dimension: progression and long-term survival. AB - PURPOSE: We present our findings in a series of patients treated with simple enucleation for RCC 4 to 7 cm in greatest dimension. We specifically report the incidence of local and systemic recurrence, and the disease specific survival rate. MATERIALS AND METHODS: We retrospectively reviewed clinical and pathological data on 71 patients who underwent nephron sparing surgery by simple enucleation between 1986 and 2004 for sporadic, unilateral, pathologically confirmed, 4 to 7 cm RCC. Patients with a solitary kidney due to previous RCC treated with radical nephrectomy were excluded from study. None of the patients had preoperative or intraoperative suspicion of positive nodes. All patients were free of distant metastases before surgery (M0). Patient status was last evaluated in May 2005. Mean followup was 74 months (median 51, range 12 to 225). RESULTS: Pathological review according to the 2002 TNM classification showed that 42% of the tumors (30 of 71) were pT1a, 44% (31 of 71) were pT1b and 14% (10 of 71) were pT3a. Mean tumor greatest dimension +/- SD was 4.7 +/- 0.81 cm (median 4.5, range 4.0 to 7.0) cm. None of the patients died within the first 30 days of surgery. There were no major complications requiring open reoperation, such as bleeding and urinary leakage/urinoma. Five and 8-year cancer specific survival was 85.1% and 81.6%, respectively. Five-year cancer specific survival in patients with pT1a (4 cm), pT1b and pT3a disease was 95.7%, 83.3% and 58.3%, respectively (pT1a vs pT1b p = 0.254, pT1a vs pT3a p = 0.006 and pT1b vs pT3a p = 0.143). Overall 10 patients experienced progressive disease (14.9%), of whom 3 had local recurrence (4.5%) alone or local recurrence associated with distant metastases. CONCLUSIONS: Simple tumor enucleation is a useful and acceptable approach to nephron sparing surgery for 4 to 7 cm RCC. It provides long-term cancer specific survival rates similar to those of radical nephrectomy and is not associated with a greater risk of local recurrence than partial nephrectomy for RCC less than 4 cm in greatest dimension. PMID- 16697793 TI - Nephron sparing surgery for renal cell carcinoma with normal contralateral kidney: 25 years of experience. AB - PURPOSE: We report the long-term results of our consecutive series of 504 patients who underwent NSS for cancer suspicious, solid renal tumors in the presence of a normal opposite kidney at our institution since 1979. MATERIALS AND METHODS: A total of 715 patients underwent NSS since 1969, including 504 for an elective indication, that is with a normal opposite kidney. Of these patients 381 (75.6%) had RCC, 123 (24.4%) had cancer suspicious benign lesions, including 53 (10.5%) with oncocytoma, 33 (6.5%) with angiomyo(lipo)ma, 23 (4.6%) with a complicated cyst and 13 (2.8%) with other benign lesions. Of the 381 patients with RCC 283 (74.3%) had clear cell, 68 (17.8%) had papillary and 30 (7.9%) had chromophobic RCC. Mean tumor diameter was 3.0 cm (range 0.5 to 11.0). Mean followup was 6.77 years (range 0.2 to 24.1). The oncological outcome was studied, including pathological features associated with tumor progression. RESULTS: Estimated cancer specific survival rates at 5 and 10 years were 98.5% and 96.7%, respectively. Estimated survival rates free of distant metastasis at 5 and 10 years were 97.5% and 95.1%, respectively. Nine patients with localized RCC experienced local recurrence after NSS. Estimated survival rates free of local recurrence at 5 and 10 years were 98.3% and 95.7%, respectively. CONCLUSIONS: The long-term results of our series support the concept of organ sparing surgery for RCC in the presence of a normal opposite kidney with excellent long-term survival and a low tumor recurrence rate. PMID- 16697794 TI - Multiplex polymerase chain reaction for microsatellite analysis of urine sediment cells: a rapid and inexpensive method for diagnosing and monitoring superficial transitional bladder cell carcinoma. AB - PURPOSE: Several urinary markers have been recently introduced in clinical practice for improving the noninvasive diagnosis of transitional cell carcinoma. Although microsatellite analysis must be considered the best method in terms of results, its cost and method time are unacceptable for daily use. We validated a more rapid and inexpensive method of determination using rapid DNA extraction and automatic multiplex polymerase chain reaction amplification. MATERIALS AND METHODS: A total of 120 patients who presented consecutively to a urological office, including 73 with transitional cell carcinoma and 43 who served as controls, were selected for study. Microsatellite analysis was performed in the blood/urine pair using 3 multiplex polymerase chain reactions per patient. Urine sediment inflammatory cells were assessed by urine dipstick test. Ten microsatellite loci were investigated. Numerical data collected during electrophoresis of the amplified segment in an ABI Prism 310 Genetic Analyzer were used to calculate the cutoff for allelic imbalance. Method sensitivity, specificity, and positive and negative predictive values were calculated. RESULTS: A total of 66 patients had microsatellite analysis alterations in urine sediment, of whom 59 had transitional cell carcinoma, while 7 had other urological diseases. Test sensitivity and specificity were 80.8% and 85.1%, respectively. Statistical analysis did not indicate any significant influence of inflammatory status on microsatellite analysis diagnostic performance. In the control group the allelic imbalance on chromosome 9 was significantly lower than on other chromosomes (p = 0.0143). This could confirm that chromosome 9 has a specific role in transitional cell carcinoma. The multiplex microsatellite analysis method was low cost and not time-consuming. CONCLUSIONS: Multiplex microsatellite analysis is a noninvasive, rapid, inexpensive and reproducible method for screening for and monitoring superficial transitional cell carcinoma. It should be considered an alternative method to urinary cytology and it should also be considered in the presence of urine sediment inflammatory cells. PMID- 16697796 TI - Association between urinary cytology and pathology for nontransitional cell malignancies of the urinary tract. AB - PURPOSE: Because nontransitional cell carcinoma neoplasms of the urinary tract are rare in Western countries, we examined the association between urinary cytology and pathology evaluations for these tumors. MATERIALS AND METHODS: An institutional review board approved, retrospective review of a total of 55,946 cytology evaluations in 12,705 patients between 1992 and 2004 was performed for correlation with subsequent histopathology findings. Documented urothelial neoplasms were then correlated with previous cytology results. Nontransitional cell carcinomas were categorized as adenocarcinoma, squamous cell carcinoma and other, including small cell disease, sarcoma, melanoma or lymphoma. RESULTS: All 108 patients with cytology evaluations showing adenocarcinoma had histological evidence of cancer and 86% had adenocarcinoma in the urinary tract. All 110 patients with squamous cell carcinoma on cytology had cancer, including 47% with primary squamous cell disease. All 42 patients with other nontransitional cell carcinomas on cytology evaluation had cancer, of whom 64% had histological concordance. In a separate analysis of 70 patients who had pathologically confirmed adenocarcinoma 57% had positive prior cytology findings, of whom 19% had histological concordance. Of 85 patients with squamous cell carcinoma 81% had positive prior cytology findings, of whom 60% had histological concordance. Of 83 patients with other nontransitional cell carcinomas 70% had positive prior cytology findings, of whom 31% had histological concordance. CONCLUSIONS: In our series all patients with nontransitional cell carcinoma cytological results had cancer in the urinary tract. Thus, nontransitional cell carcinoma cytology findings mandate careful urinary tract evaluation. Concordance with histological subclassification on subsequent pathology evaluation ranges from 49% for squamous cell carcinoma to 86% for adenocarcinoma. A majority of patients with nontransitional cell carcinoma malignancies had positive prior cytology findings. However, the concordance with histological subclassification on prior cytology results ranges from 19% for adenocarcinoma to 60% for squamous cell carcinoma. PMID- 16697798 TI - Population based survival data on urachal tumors. AB - PURPOSE: Urachal carcinoma accounts for less than 1% of all bladder cancers. Limited data exist on disease related outcomes originating from case reports and select referral centers. We describe a population based outcomes analysis with long-term followup in patients in the province of Ontario. MATERIALS AND METHODS: We reviewed the data source of the Ontario Cancer Registry for patients diagnosed with urachal cancer during 1976 to 2001. A cohort of 40 patients with urachal adenocarcinoma was found. Primary outcome measures were overall and disease specific survival. The effect of patient age, sex, grade, stage and university vs nonacademic treating hospital as predictors of outcome was determined. RESULTS: Median patient age was 52 years. Median followup was 72.7 months. Mean overall survival +/- SD was 121.6 +/- 21 months. Mean disease specific survival in patients treated operatively was 165 +/- 27 months with 5 and 10-year disease specific survival of 61.3% and 49.2%, respectively. Disease specific mortality was not evident after 7 years from diagnosis. Well differentiated tumors in a third of the patients were associated with a 90% cure rate when treated operatively. Well differentiated tumors, and noninvolvement of adjacent organs and the peritoneum correlated with better prognosis (p = 0.004, p = 0.03 and 0.045, respectively). CONCLUSIONS: Urachal adenocarcinoma occurs in all age groups. Long-term disease specific survival can be achieved with partial cystectomy. Covariates associated with better disease specific survival are well differentiated tumor grade and the absence of adjacent organ or peritoneal involvement. No relapses were observed after 7 years. PMID- 16697800 TI - Clinical outcomes following radical cystectomy for primary nontransitional cell carcinoma of the bladder compared to transitional cell carcinoma of the bladder. AB - PURPOSE: The effect of bladder cancer histological subtypes other than transitional cell carcinoma (nonTCC) on clinical outcomes remains uncertain. We conducted a multi-institutional retrospective study of patients with bladder cancer treated with radical cystectomy to assess the impact of nonTCC histology on bladder cancer specific outcomes. MATERIALS AND METHODS: A total of 955 consecutive patients underwent radical cystectomy with bilateral pelvic lymphadenectomy for bladder cancer at 3 academic institutions. TCC was present in the radical cystectomy specimen in 888 patients (93%). NonTCC histology was present in 67 patients (7%), including squamous cell carcinoma in 26, adenocarcinoma in 13, small cell carcinoma in 10 and other nonTCC subtypes (ie spindle cell carcinoma, carcinosarcoma and undifferentiated carcinoma) in 18. For patients alive at last followup median followup was 39 and 23 months for patients with TCC and nonTCC histologies, respectively. Bladder cancer specific progression and survival were assessed using Kaplan-Meier and multivariate Cox proportional hazards analyses. RESULTS: Bladder cancer specific progression and mortality did not differ significantly between patients with SCC and TCC histologies. Patients with nonTCC and nonSCC bladder cancer were at significantly increased risk for progression and death compared to patients with TCC or SCC (p <0.001). This association remained statistically significant in patients with organ confined disease (stage pT2 or lower) and patients with nonorgan confined disease (stage pT3 or higher) (p <0.001). In a multivariate analysis nonTCC and nonSCC histology was associated with an increased risk of bladder cancer progression and death (OR 2.272 and 2.585, respectively, p <0.001), even after adjusting for final pathological stage, lymph node status, lymphovascular invasion and neoadjuvant or adjuvant treatments. CONCLUSIONS: NonTCC and nonSCC histological subtype is an independent predictor of bladder cancer progression and mortality in patients undergoing radical cystectomy for bladder cancer. Patients with bladder TCC and SCC share similar stage specific clinical outcomes. PMID- 16697802 TI - Patterns of initial transitional cell recurrence in patients after cystectomy. AB - PURPOSE: We characterized initial relapse patterns in patients who underwent cystectomy for transitional cell carcinoma of the bladder at our institution. MATERIALS AND METHODS: Between June 1995 and December 2001 a total of 417 patients underwent radical cystectomy for transitional cell carcinoma. Of the 399 patients treated with orthotopic diversion or incontinent cutaneous diversion, recurrent TCC developed in 86 (21.6%). We retrospectively examined this population to determine if any perioperative variables influenced the first site of disease recurrence. RESULTS: Of the 86 patients in whom metastatic disease developed after cystectomy, the mean followup was 22.1 months. Mean time to disease recurrence was 15.1 months. Recurrence was most commonly initially in the pelvis (30.2%). Other common sites of first recurrence included the viscera (26.7%), the retroperitoneum (16.3%) and the skeletal system (16.3%). The site of first recurrence did not correlate significantly with gender, age, diversion type, pathological stage, nodal status, prostatic involvement or the presence of a positive surgical margin. Lymphovascular invasion was more prevalent in the pelvic recurrence group (62%) than in the retroperitoneal (36%), visceral (22%) or skeletal metastases (29%) groups (p = 0.03). CONCLUSIONS: While the pelvis is the most common site of disease recurrence following radical cystectomy, TCC of the bladder recurs in an unpredictable pattern and aggressive surveillance is indicated. Patients with local pelvic recurrence had a higher percentage of lymphovascular invasion on cystectomy than the other recurrence site groups. PMID- 16697803 TI - Partial cystectomy for muscle invasive urothelial carcinoma of the bladder: a contemporary review of the M. D. Anderson Cancer Center experience. AB - PURPOSE: Partial cystectomy is a surgical option for select patients diagnosed with urothelial carcinoma. We review our experience with partial cystectomy for muscle invasive urothelial carcinoma to assess local control and survival rates. MATERIAL AND METHODS: From 1982 to 2003 a total of 37 patients with muscle invasive urothelial carcinoma underwent partial cystectomy with curative intent. Reviewed data included history of superficial tumors, presence of variant histology, tumor location, clinical stage, pathological stage, presence of carcinoma in situ, adjuvant therapy and disease status. RESULTS: The 5-year overall, disease specific and recurrence-free survival rates were 67%, 87% and 39%, respectively. Mean followup was 72.6 months (range 6 to 217). Of the 37 patients 19 (51%) did not have tumor recurrence, 9 (24%) had superficial recurrence in the bladder that was treated successfully and 9 (24%) had recurrence with advanced disease. A total of 24 patients (65%) had an intact bladder with no evidence of disease after a median of 53 months. There were 6 patients (16%) who died of bladder cancer, 3 of whom died of late recurrence of muscle invasive cancer (41, 44 and 138 months after partial cystectomy). On multivariate analysis higher pathological stage (HR 3.4, p = 0.04) was associated with shorter recurrence-free survival. A history of superficial tumors (p <0.01) and clinical stage (p = 0.01) was associated with advanced recurrence-free survival. The use of adjuvant chemotherapy (HR 0.18, p = 0.03) was associated with prolonged advanced recurrence-free survival, however adjuvant chemotherapy did not impact overall survival. CONCLUSIONS: Partial cystectomy provides adequate local control of muscle invasive bladder cancer in select patients. Because late recurrence is not uncommon and is potentially life threatening, lifelong followup with cystoscopy is recommended. PMID- 16697804 TI - Urothelial carcinoma after external beam radiation therapy for prostate cancer. AB - PURPOSE: We reviewed the clinical course of patients in whom urothelial carcinoma developed following radiation therapy for prostate cancer. MATERIALS AND METHODS: A retrospective review of all patients between 1990 and 2005 with the diagnosis of bladder and prostate cancer was performed. Of 125 total patients new onset urothelial carcinoma developed in 11 after undergoing external beam radiation therapy for prostate cancer. RESULTS: Whole pelvis external beam radiation therapy with a proton boost to the prostate was the radiation modality in 7 of the 11 patients (64%), while the remaining 4 patients received standard external beam radiation only. Urothelial carcinoma was detected a mean of 3.07 years after completion of radiation therapy in the proton group, compared to a mean latency period of 5.75 years in the standard radiation group (p = 0.09). Average patient age at diagnosis was 72 years (range 64 to 84). All patients presented with gross hematuria and had cystoscopic findings of coexisting radiation cystitis. Of the 11 patients 5 (45%) presented with grade 3 carcinoma and eventually 7 (64%) required radical cystectomy. Urothelial tumors with sarcomatoid features (carcinosarcoma and spindle cell sarcomatoid) developed in 2 patients (18%). Of the 11 patients 10 (91%) were nonsmokers at the time of urothelial carcinoma diagnosis. CONCLUSIONS: Urothelial carcinoma in patients with previous radiation therapy for prostate cancer is often high grade, and the majority of patients have cancer progression requiring cystectomy. A high incidence of urothelial carcinoma with sarcomatoid features was seen in these patients. PMID- 16697805 TI - Expression and cellular localization of follicle-stimulating hormone receptor in normal human prostate, benign prostatic hyperplasia and prostate cancer. AB - PURPOSE: FSH, identified as an endogenous product of the prostate, is a glycoprotein with proliferative activity. Increasing evidence of autocrine/paracrine activities of gonadotropins at extragonadal sites led us to investigate the gene expression and cellular localization of FSH-R in normal and diseased human prostates. MATERIALS AND METHODS: Prostate specimens, including normal gland, BPH, PCa and human androgen refractory (PC3) and androgen dependent (LNCaP) prostate cancer cell lines (European Collection of Cell Cultures, Salisbury, United Kingdom), were analyzed for FSH-R expression by semiquantitative and real-time reverse transcriptase-polymerase chain reaction and immunohistochemistry. We also evaluated cyclic adenosine monophosphate production by cultured PC3 and LNCaP stimulated with human FSH. RESULTS: Little FSH-R expression was seen in 9 of 13 normal and 8 of 15 BPH specimens. Of 30 PCa samples 21 were FSH-R positive with generally higher expression compared to normal prostate and BPH samples. Real-time reverse transcriptase-polymerase chain reaction of matched normal/tumor pairs confirmed higher FSH-R mRNA expression in PCa. PC3 cells expressed FSH-R, while LNCaP cells were FSH-R negative. FSH-R protein was mainly localized in the glandular epithelium and in some stromal cells in normal prostate, BPH and PCa specimens. PC3 cells expressed FSH-R protein and their treatment with FSH induced a significant increase in cyclic adenosine monophosphate production. CONCLUSIONS: These results indicate that a subset of PCa expresses FSH-R mRNA and protein at levels higher than those of normal and hyperplastic tissues that express FSH-R. This suggests that FSH might contribute to some cases of PCa via a receptor mediated mechanism. PMID- 16697807 TI - Prevalence of prostate specific antigen testing for prostate cancer in elderly men. AB - PURPOSE: We investigated the prevalence and outcome of PSA testing for prostate cancer screening or diagnosis in elderly men 75 years or older at our academic medical center. MATERIALS AND METHODS: A cross-sectional study design was used to identify all men 75 years or older who underwent a PSA test through the family medicine or internal medicine service at our institution between January 1, 1998 and June 30, 2004. All patients with a suspected (PSA less than 0.1 ng/ml) or confirmed prior diagnosis of prostate cancer were excluded. The prevalence of PSA testing was then compared to that in younger age groups (45 to 54, 55 to 64 and 65 to 74 years). We then examined the frequency and nature of further evaluation and treatment performed in men following the PSA test. RESULTS: The 8,787 male patients who were 75 years or older generated a total of 82,672 visits in the 5.5 year period. Of these patients 505 (5.7%) underwent at least 1 PSA test. The prevalence of PSA testing in the younger age groups was 10.3% (1,769 of 17,175) in patients 45 to 54 years old, 14.9% (2,052 of 13,772) in those 55 to 64 years old and 11.8% (1,258 of 10,661) in those 65 to 74 years old (chi-square test p <0.001). Of these patients 98 of 343 (28.6%) with PSA between 0.1 and 4 ng/ml were referred to a urologist at our institution and 3 underwent biopsy. None had a prostate cancer diagnosis. Of the 162 patients with PSA more than 4 ng/ml 84 (51.9%) were referred to a urologist. Only 10 of the 84 patients (11.9%) who were referred to a urologist underwent prostate biopsy. Six of the 10 men (60%) were diagnosed with prostate cancer, including 1 with a Gleason 6 tumor, 1 with a Gleason 7 tumor and 4 who were found to have tumors with a Gleason score of 8 or greater. All patients received androgen deprivation therapy, except 1 who received local external beam radiation therapy. An additional patient was diagnosed by biopsy of a vertebral lesion and he received hormone therapy. At a median followup of 51 months (range 28 to 72) 4 of 7 men (57%) were alive with disease. CONCLUSIONS: PSA testing for prostate cancer screening and diagnosis appear to decrease with advancing age. A small but significant proportion of men who are 75 years or older continue to undergo PSA testing. Abnormal PSA results do not always result in further evaluation and therapy for prostate cancer in elderly men. The establishment of firm guideline recommendations regarding PSA testing and further evaluation for prostate cancer in elderly men, perhaps based on individualized geriatric assessment, may be helpful. PMID- 16697808 TI - Suprapubic prostatectomy with a novel catheter. AB - PURPOSE: We evaluated the postoperative morbidity and technical complications of a new handmade catheter used for suprapubic prostatectomy. MATERIALS AND METHODS: A total of 146 patients with lower urinary tract symptoms who had indications for open prostatectomy were enrolled in the study from January 2003 to August 2004. Of the patients 96 were treated with a novel technique without a urethral catheter but with a special handmade cystostomy catheter, while the other 50 underwent the traditional procedure using cystostomy and a urethral catheter. The same surgical team operated on the 2 groups. Clot retention episodes, hemoglobin decreases, irritative symptoms, voiding status after cystostomy removal and incontinence were evaluated postoperatively. Three months later we followed the patients with symptom score, maximum flow rate and cystoscopic examination. RESULTS: Preoperatively mean International Prostate Symptom Score was 31.6 in each group. Postoperatively none of the patients with the novel catheter complained of significant irritative urinary symptoms, clot retention and true or stress urinary incontinence. Of the cohorts 94% were satisfied with voiding but 86% of controls were satisfied. There was no report of urinary tract infection or epididymo-orchitis in the cohorts, while we found epididymo-orchitis in 4 controls (8%). Three months after the operation the mean International Prostate Symptom Score was 4.4 (range 1 to 7) and the mean maximum flow rate was 22.6 ml per second (range 14 to 25) in patients with the novel catheter, and 4.2 (range 1 to 7) and 22.5 ml per second (range 15 to 25), respectively, in those with the traditional catheter. At followup there was no bladder neck contracture but 4 patients (4.1%) showed some degree of membranous urethral stricture. We also noted 7 controls (14%) with urethral stricture. CONCLUSIONS: Transurethral prostate resection has been introduced as the surgical treatment of choice in patients with benign prostatic hyperplasia. However, open adenomectomy still has a place. Urethral catheter-free suprapubic prostatectomy can be safely applied with a low postoperative risk of infection, incontinence and stricture formation. PMID- 16697809 TI - Clinical and immunological characteristics of patients with serologic progression of prostate cancer achieving long-term disease control with granulocyte macrophage colony-stimulating factor. AB - PURPOSE: We describe the clinical and immunological characteristics of patients with biochemically relapsed prostate cancer who achieved long-term disease control with GM-CSF (Leukine). MATERIALS AND METHODS: A total of 30 patients with prostate cancer and nonmetastatic recurrent disease, as manifested by increasing PSA between 0.4 and 6.0 ng/ml after prior definitive therapy, were enrolled in a phase II trial. Patients received 250 microg/m2 GM-CSF daily subcutaneously on days 1 through 14 of a 28-day cycle until PSA or objective progression. The patient and disease characteristics of patients who remained without evidence of disease progression beyond 4 years were examined. Additionally, flow cytometry was performed in peripheral blood to characterize monocyte and dendritic cells. RESULTS: Seven of 29 evaluable patients (24%) remained free of disease progression at a median of 5.1 years (range 4.5 to 5.6 or greater) from the start of GM-CSF therapy. Patients on long-term GM-CSF tended to have lower initial T stage, Gleason score and pretreatment PSA. An increase in the number of circulating monocytes and dendritic cells was observed after 14 days of GM-CSF treatment. These values returned to baseline during the 14-day off period. CONCLUSIONS: GM-CSF modulates PSA in androgen dependent, biochemically relapsed cases. A substantial proportion of patients achieve long-term disease control. The clinical characteristics described may help select patients for future clinical trials with GM-CSF or other immunomodulators. Additional investigation is required to define the immunological mechanism of GM-CSF in prostate cancer. PMID- 16697810 TI - The impact of neoadjuvant hormonal therapy on the outcome of laparoscopic radical prostatectomy: a matched pair analysis. AB - PURPOSE: We evaluated the effect of androgen ablation treatment on laparoscopic radical prostatectomy operative and postoperative parameters. MATERIALS AND METHODS: A total of 50 patients (group 1) on neoadjuvant androgen deprivation, followed by laparoscopic radical prostatectomy, were compared to 50 (group 2) without any treatment who were matched for prostate volume, laparoscopic pelvic lymphadenectomy, nerve sparing procedure, surgical access type and pathological stage. We analyzed operative time, blood loss, intraoperative and postoperative complications, catheter time, procedure difficulty as scored by the surgeon and surgical margin status. RESULTS: There was no significant difference between the neoadjuvant and nonneoadjuvant groups with respect to mean operative time +/- SD (228.6 +/- 62.9 vs 219.4 +/- 65.1 minutes), mean blood loss (667.6.1 +/- 217.1 vs 729.8 +/- 285.1 ml) and median catheter time (7 vs 7.5 days). We also found no difference related to the complication rate. Ten of 50 prostate dissections (20%) in group 1 were classified as difficult, whereas in group 2 only 4 of 50 (8%) were scored as difficult (p = 0.084). The positive surgical margin rates did not differ. CONCLUSIONS: There was no significant difference with respect to operative or postoperative parameters in patients undergoing neoadjuvant androgen ablation therapy compared to controls. At centers where there is experience laparoscopic radical prostatectomy can be safely performed in patients who have undergone neoadjuvant hormonal therapy. PMID- 16697811 TI - Long-term benefits of elective radiotherapy after prostatectomy for patients with positive surgical margins. AB - PURPOSE: The benefit of adjuvant radiotherapy after prostatectomy for patients with pathological risk factors but with an undetectable postoperative PSA remains controversial. In this retrospective study we define the benefits of elective postoperative radiotherapy in this setting. MATERIALS AND METHODS: A total of 44 patients received elective postoperative radiotherapy at a single institution in the PSA era (1989 to 1995) for positive surgical margins and undetectable postoperative PSA. Radiotherapy was delivered to a median dose of 60 Gy. Clinical target volume included the prostate bed. Pelvic nodes were not treated. The four field box technique with customized blocking of bladder, rectum and small bowels was used and defined the planning target volume. The patients were then compared to a contemporaneous group of 189 patients with positive surgical margins who underwent radical prostatectomy without any adjuvant therapy. Failure was defined as biochemical (PSA) recurrence and was timed from first detectable PSA. RESULTS: The 5 and 10-year biochemical no evidence of disease was 90.9% and 90.9% for the elective postoperative radiotherapy group, and 66.4% and 54.5% for the observation group, respectively (p = 0.0012). Median time to biochemical failure was also longer in the elective postoperative radiotherapy group (88.6 months) compared to the observation group (43.5 months) (p <0.001). Risk factors for biochemical recurrence on multivariate analysis were Gleason score greater than 7 (p = 0.017), established extracapsular extension (p = 0.002) and lack of elective postoperative radiation (p = 0.001). CONCLUSIONS: This is one of the longest followup studies showing that elective postoperative radiation therapy is associated with improved bNED and prolonged time to recurrence. Combined radical prostatectomy and elective postoperative radiotherapy should be considered in the management of high risk prostate cancer, especially in the presence of positive surgical margins despite undetectable PSA. PMID- 16697813 TI - Nomogram predictive of cancer specific survival in patients undergoing partial or total amputation for squamous cell carcinoma of the penis. AB - PURPOSE: We created the first nomograms to predict cancer specific survival probabilities of patients with squamous cell carcinoma of the penis, clustering prognostic information from the most commonly used clinical and pathological variables. MATERIALS AND METHODS: We retrospectively collected clinical and pathological data from 175 patients who had undergone surgery for squamous cell carcinoma of the penis from 1980 to 2002 at 11 urological centers in northeastern Italy. A logistic regression model was used to construct the nomogram. RESULTS: At a median followup of 24 months, 101 patients (57.7%) were alive and disease free while 74 (42.3%) died of penile cancer. According to multivariate analyses, 2 models predictive of cancer specific survival probability were generated. The first model was based on the pathological findings of the primary tumor after penectomy and on the clinical stage of groin lymph nodes, while the second model included the pathological data of the primary tumor and groin lymph nodes. The concordance index was 0.728 for the first model and 0.747 for the second. Calibration appeared to be good in both models. CONCLUSIONS: In this article we propose 2 models to predict the 5-year cancer specific survival probabilities of patients with squamous cell carcinoma of the penis. Both models showed good discriminating power and calibration in predicting patient 5-year cancer specific survival. These nomograms could improve the quality of prognostic data provided to patients and support physicians in planning treatment. PMID- 16697815 TI - An analysis of the natural history of Peyronie's disease. AB - PURPOSE: Little information exists on the natural history of PD. We defined the course of PD in a group of men with this condition who received no treatment. MATERIALS AND METHODS: The study population comprised patients with PD who presented within 6 months of disease onset, had no medical treatment and were followed until at least 12 months after disease onset. At baseline and followup penile abnormality was determined following intracavernous injection and by measurement at maximum penile rigidity. RESULTS: A total of 246 patients met inclusion criteria. At presentation mean age +/- SD was 52 +/- 22 years and the duration of PD was 3.5 +/- 1.5 months. At baseline in men with documented curvature 72% had dorsal, 17% had ventral and 11% had lateral curvature. Mean curvature at baseline was 42 +/- 22 degrees. Mean stretched flaccid penile length was 12.2 cm. The mean duration of PD at the followup assessment was 18 +/- 7 months. At followup stretched flaccid length had decreased to 11.4 cm (p = 0.035). Of the patients 32% complained of some degree of erectile dysfunction at baseline. All patients who reported penile pain had improvement and 89% reported complete resolution at followup. Of men with curvature 12% had improved, 40% remained stable and 48% had worsened at followup. In those in whom curvature improved the mean change was 15 degrees, while in those in whom curvature worsened the mean change was 22 degrees. CONCLUSIONS: To our knowledge this is the largest study to explore the natural history of PD. A minority of men experienced improvement in penile abnormality, while penile length decreased during the 1-year followup. This information will permit clinicians to provide patients with realistic expectations at presentation for the evaluation of PD. PMID- 16697817 TI - Genetic heritability of urinary stone risk in identical twins. AB - PURPOSE: Quantitative measurements of urinary parameters are valuable clinical tools for predicting the risk of nephrolithiasis. To our knowledge no previous studies have evaluated the heritability of urinary stone risk in identical twins. Because these individuals share identical genetics, the R defined by their phenotypic data are theoretically equal to the entire population H2. MATERIALS AND METHODS: A total of 12 sets of healthy homozygous twins, including 4 males and 8 females for a total of 24 individuals, with a mean age of 25.9 years (range 21 to 36) volunteered for this study. All subjects provided informed written consent before assessment. Urinary stone risk profiles were done elsewhere on 2 consecutive days after 5 days of a standardized diet (170 mEq Na and 2500 kcal). Linear regression was performed on the data to determine R. Because identical twins were used, R was theoretically an estimate of H2. RESULTS: Certain urinary stone risk markers were highly heritable, including urinary calcium (94%), oxalate (94%), citrate (95%), uric acid (96%) and brushite supersaturation (90%), as determined by genotype (H2 90% or greater). Uric acid supersaturation (58%) and urinary sodium (64%) had low degrees of heritability. CONCLUSIONS: H2 is a measure of how much of the total variance in phenotype results from differences in genotype, as opposed to environmental differences. For example, an H2 of 95% for citrate suggests that genetic differences account for 95% of the variation in urinary citrate and environmental differences account for the remaining 5%. Therefore urinary calcium, oxalate and citrate are primarily determined by genotype, while environmental factors, particularly those that impact urine pH and urinary volume, may be increasingly important for determining uric acid supersaturation. PMID- 16697818 TI - Ureteroscopic laser lithotripsy for upper urinary tract calculi with active fragment extraction and computerized tomography followup. AB - PURPOSE: Management of fragments generated by ureteroscopic laser lithotripsy remains controversial. In this study we explored the impact of active fragment extraction after ureteroscopic laser lithotripsy on stone clearance. MATERIALS AND METHODS: A total of 69 patients with 3 or less upper urinary tract calculi (5 to 15 mm) demonstrated on preoperative CT were prospectively evaluated. Stones were translocated to a dependent upper pole calix where laser lithotripsy was performed. An attempt was made to clear all fragments using tipless stone baskets. One month after surgery stone clearance was evaluated exclusively with noncontrast spiral CT. RESULTS: In 58 patients undergoing surgery on protocol, average stone burden was 9.4 +/- 3.4 mm and was significantly smaller in 44 patients with stones in a solitary location (8.5 +/- 2.9 mm) than in 14 patients with stones in multiple locations (12.3 +/- 3.2 mm, p <0.001). Primary stone location was categorized as renal nonlower pole (in 16), renal lower pole (in 19) and proximal ureter (in 23). Average operative time (43.7 +/- 18.4 minutes) was unaffected by stone location or multiplicity after controlling for stone size (p >0.05). Stone clearance rates were not affected by stone location or multiplicity, with overall success rates of 54%, 84% and 95% at fragment thresholds of 0, 2 and 4 mm, respectively (p >0.05). CONCLUSIONS: Ureteroscopic laser lithotripsy with active fragment extraction was time efficient and highly effective. Sensitive postoperative imaging reveals the challenge of achieving a true stone-free state. We were unable to demonstrate an impact of stone location on stone-free rates. PMID- 16697820 TI - Exploration for hemorrhage following laparoscopic renal surgery: intraoperative findings. AB - PURPOSE: Bleeding after surgery is a rare but potentially life threatening complication. We reviewed operative and postoperative clinical features in patients who required surgical exploration secondary to hemorrhage following laparoscopic renal procedures. MATERIALS AND METHODS: We retrospectively reviewed the records of patients undergoing laparoscopic renal surgery between January 1996 and September 2004. Nine of 1,123 patients (0.8%) underwent early exploration for bleeding within 5 days of surgery. RESULTS: Two groups were identified. Group 1 consisted of 4 patients who underwent early exploration at less than 10 hours after surgery and had arterial bleeding. Group 2 consisted of 5 patients who underwent exploration a mean 38 hours after surgery and in whom no bleeding source was identified. Group 1 patients had pronounced hypotension with systolic blood pressure 70 to 79 mmHg and hematocrit decreases (mean 10.5%) in a short time course before repeat exploration (mean 4.5 hours). Arterial bleeding was identified in the hilum and adrenal bed. Group 2 patients demonstrated a decrease in hematocrit from an initial mean of 28.3% to 22.5% with tachycardia and mild hypotension (systolic blood pressure 90 to 99 mmHg). On exploration group 2 patients had diffuse oozing. Mean hospital stay in group 1 was 8 days (range 4 to 9) vs 12 (range 6 to 24) in group 2. CONCLUSIONS: Early hemodynamic instability after laparoscopic renal surgery is likely to indicate a discrete arterial bleeding source from the hilum or adrenal bed, requiring surgical control. In patients who underwent exploration after a delayed bleeding presentation no discrete source was found intraoperatively. Therefore, it is unclear whether these patients benefited from surgical exploration. PMID- 16697821 TI - Urethral and bladder neck injury associated with pelvic fracture in 25 female patients. AB - PURPOSE: We describe the presentation, diagnostic evaluation, management and outcome of female urethral trauma. MATERIALS AND METHODS: All female patients treated at Harborview Medical Center between 1985 and 2001 with urethral injury were identified by International Classification of Diseases 9th revision code. Approval of the Human Subject Division was obtained and patient charts were reviewed. The Urogenital Distress Inventory Short Form, the Incontinence Impact Questionnaire Short Form and the Female Sexual Function Index were sent to the patients. RESULTS: A total of 25 patients (13 adults, 12 children) with a mean age of 22 years (range 4 to 67) met inclusion criteria. All had pelvic fracture related to blunt trauma. They represented 6% of all female patients treated in the same review period with pelvic fracture. Blood was seen at the introitus in 15 patients and 19 had gross hematuria. Of the injuries 9 were avulsions, 15 were longitudinal lacerations and 1 was not further specified. Primary repair was performed in 21 patients and 4 were treated nonoperatively. There were 5 patients who required secondary procedures including fistula repair in 4 and continent urinary diversion in 1. At a mean followup of 7.3 years (range 1.6 to 14.4) 9 of 21 patients (43%) had moderate or severe lower urinary tract symptoms and 8 of 13 (38%) had sexual dysfunction (FSFI score less than 26.55). CONCLUSIONS: Female urethral and bladder neck injury occurs with pelvic fracture, presents with gross hematuria and/or blood at the introitus, and requires operative repair for avulsions and longitudinal lacerations. These patients are at risk for significant sexual and lower urinary tract dysfunction. PMID- 16697823 TI - Proximal bulbar urethroplasty via extended anastomotic approach--what are the limits? AB - PURPOSE: We report our initial experience with men who underwent EAU for strictures greater than 2.5 cm involving the proximal bulbar urethra. MATERIALS AND METHODS: Of the more than 250 men who underwent urethral reconstruction at our institution during 1997 to 2005 a select consecutive group of 22 in whom proximal bulbar urethral strictures were treated with primary bulbomembranous anastomosis were evaluated. Outcomes in men with strictures greater than 2.5 cm long (EAU) were compared to those in men with shorter strictures in the same proximal bulbar location. Cases of post-traumatic urethral disruption related to pelvic fractures were omitted. American Urological Association symptom index scores and erectile function questionnaires were completed more than 6 months postoperatively. Results of a prior study using the same erectile function questionnaire after various types of urethroplasty and circumcision were then compared to those of our series. RESULTS: Patients with EAU had an average stricture length of 3.78 cm (range 2.6 to 5.0) and 10 of 11 procedures (91%) were successful. Anastomotic urethroplasty performed for similar proximal bulbar strictures less than 2.5 cm (mean 1.5, range 1.0 to 2.3) was successful in 10 of 11 cases (91%). Mean followup was 22.1 months and all followups were more than 1 year. Men treated with EAU had no increased rate of stricture recurrence or erectile complaints compared to men in whom shorter proximal bulbar strictures were repaired using an identical surgical technique. Similarly no increased rate of erectile problems was identified compared to other types of urethroplasty and circumcision using an identical questionnaire. CONCLUSIONS: Urethral reconstructability is proportional to the length and elasticity of the distal urethral segment. Defects up to 5 cm may be successfully excised and primarily reconstructed in select young men with proximal bulbar strictures. PMID- 16697825 TI - Reconstructive surgery for the management of genitourinary tuberculosis: a single center experience. AB - PURPOSE: We evaluated the role of surgery for genitourinary tuberculosis with special emphasis on reconstructive procedures. MATERIALS AND METHODS: Case records of 241 patients with genitourinary tuberculosis who underwent surgery at our center during a period of 17 years were reviewed. Clinical features, organ involvement, investigations, treatment and outcome of therapy were studied. RESULTS: There were 129 males and 112 females with a mean age of 34.6 years. The most common presentation was irritative voiding symptoms. Azotemia was seen in 54 (22.4%) cases. The most commonly involved organ was the kidney in 130 (53.94%) cases. Preoperative bacteriologic diagnosis was confirmed in 70 (29%) cases. All patients received antitubercular drug therapy for 9 months. A total of 248 procedures, including 33 endoscopic, 87 ablative and 128 reconstructive, were performed with some patients requiring more than 1 procedure. Early complications, which mainly involved the bowel, were seen in 19 (7.88%) cases. Bacteriologic cure was achieved in all culture positive cases. Renal functional parameters stabilized or improved in 44 of 54 patients (81.5%) in whom they were deranged at presentation. CONCLUSIONS: Genitourinary tuberculosis is common in developing countries. Diagnosis is often delayed because of late presentation and many patients present with cicatrization sequelae. A combination of antitubercular drug therapy and judicious surgery achieves satisfactory results in the majority of cases. With improved antitubercular drug therapy and experience with the use of bowel segments in the urinary tract, more reconstructive procedures are being performed with satisfactory outcomes. In patients who undergo reconstructive procedures, a rigorous and prolonged followup is necessary. PMID- 16697827 TI - Early and late urodynamic assessment of ileal orthotopic bladder substitutes combined with an afferent tubular segment. AB - PURPOSE: Limited information is available concerning changes in the urodynamic characteristics of orthotopic bladder substitutes with time. Therefore, we compared early and late urodynamic results in patients with an ileal orthotopic bladder substitute combined with an afferent tubular segment. MATERIALS AND METHODS: Of 139 patients surviving at least 5 years after cystoprostatectomy and ileal orthotopic bladder substitution with an afferent tubular segment 119 underwent urodynamic assessment, including 66 at a median of 9 months (early) and 77 at a median of 62 months (late). Of these patients 24 were assessed at each time point. Simultaneously all patients were asked to complete a bladder diary and questionnaire regarding continence for at least 3 days in the week preceding the urodynamic study. RESULTS: Urodynamic parameters were comparable in patients who were evaluated early and late postoperatively. In addition, median values at early and late urodynamic evaluation in the 24 patients with the 2 examinations showed no statistically significant differences for volume at first desire to void (300 vs 333 ml, p = 0.85), pressure at first desire to void (12 vs 13 cm H2O, p = 0.57), maximum cystometric capacity (450 vs 453 ml, p = 0.84), end filling pressure (19 vs 20 cm H2O, p = 0.17), reservoir compliance (25 vs 28 ml/cm H2O, p = 0.58) or post-void residual urine volume (5 vs 15 ml, p = 0.27). CONCLUSIONS: Urodynamic results after 5 years of living with an ileal orthotopic bladder substitute with an afferent tubular segment show grossly unchanged urodynamic characteristics. Patients maintain a reservoir capacity and micturition pattern consistent with a normal life-style. Reservoir pressure remained low, thereby protecting and preserving upper tract function. To achieve these results patients must be regularly followed, and the causes of bacteriuria, increased post-void residual urine and bladder outlet obstruction must be recognized and dealt with accordingly. PMID- 16697829 TI - Laparoscopic pyelopyelostomy for retrocaval ureter without excision of the retrocaval segment: first report of 6 cases. AB - PURPOSE: We report our series of 6 cases of retrocaval ureter that were successfully treated with a laparoscopic approach. MATERIALS AND METHODS: Three men and 3 women with a mean age of 31 years (range 16 to 50) were referred to our department with a diagnosis of retrocaval ureter. One patient had a 12 mm renal pelvic calculus. A transperitoneal laparoscopic approach was used in all cases. The retrocaval segment along with some of the more proximal and distal segments of the ureter was readily mobilized. The distal part of the dilated renal pelvis was transected and the divided distal segment was repositioned to lie anterior to the inferior vena cava. The renal pelvic stone in 1 patient was readily removed at this point. Re-anastomosis was then performed over a Double-J stent placed intraoperatively using 2 rows of running 4-zero polyglactin sutures with intracorporeal knot tying. The stent was removed 4 weeks later. Excretory urography was performed 3 months after the laparoscopic procedure. RESULTS: Mean operative duration was 180 minutes (range 150 to 210). Patients were discharged home at a mean of 4 days (range 3 to 5). Followup excretory urogram 3 months after laparoscopic repair revealed a widely patent anastomosis with considerable improvement in hydronephrosis in all patients. CONCLUSIONS: To our knowledge this is the largest series of laparoscopic repair of retrocaval ureter reported to date. Laparoscopic pyelopyelostomy for retrocaval ureter without excision of the retrocaval segment is associated with an excellent outcome, minimal postoperative morbidity, short hospital stay and highly satisfactory cosmetic results. PMID- 16697831 TI - Ultrasound assessment of detrusor thickness in men-can it predict bladder outlet obstruction and replace pressure flow study? AB - PURPOSE: We estimated the diagnostic accuracy of ultrasound detrusor thickness measurement for BOO and investigated whether this method can replace PFS for the diagnosis of BOO in some patients with lower urinary tract symptoms. MATERIALS AND METHODS: Detrusor thickness was measured by linear ultrasound (7.5 MHz) at a filling volume of greater than 50% of cystometric capacity in 102 men undergoing PFS for LUTS. All patients with prior treatment for bladder outlet obstruction and those with underlying neurological disorders were excluded from analysis. Detrusor thickness was correlated with PFS data. Obstruction was defined according to the Abrams-Griffiths nomogram. RESULTS: Detrusor thickness was significantly higher (p <0.0001) in obstructed (61 cases, median detrusor thickness 2.7 mm, IQR 2.4 to 3.3) compared to unobstructed (18 cases, median detrusor thickness 1.7 mm, IQR 1.5 to 2) as well as equivocal (23 cases, median detrusor thickness 1.8 mm, IQR 1.5 to 2.2) cases. A weak to medium Spearman correlation was found between detrusor thickness and PFS parameters. For a diagnosis of BOO, detrusor thickness of 2.9 mm or greater had a positive predictive value of 100%, a negative predictive value of 54%, specificity of 100% and sensitivity of 43%. ROC analysis revealed that detrusor thickness had a high predictive value for BOO with an AUC of 0.88 (95% CI 0.81-0.94). CONCLUSIONS: In men with LUTS without prior treatment and/or neurological disorders, ultrasonographically assessed detrusor thickness 2.9 mm or greater has a high predictive value for BOO and can replace PFS for the diagnosis of BOO. However, this cutoff value needs to be validated in a larger study population. PMID- 16697832 TI - Interrater reliability of filling cystometrogram interpretation in a multicenter study. AB - PURPOSE: As part of a continuous quality control effort to measure the interrater reliability of urodynamic studies performed at multiple centers, we compared agreement levels for urodynamic studies between central and local physician reviewers. We report interrater reliability findings for the filling cystometrogram. MATERIALS AND METHODS: Following a satisfactory interrater reliability study among 4 central physician reviewers in 33 tracings 36 urodynamic study tracings from 9 Urinary Incontinence Treatment Network continence treatment centers and 13 Urinary Incontinence Treatment Network certified urodynamic study testers were randomly selected for review. These tracings were originally interpreted by 11 local physician reviewers using standardized Urinary Incontinence Treatment Network interpretation guidelines. Each of the 4 central physician reviewers reviewed 9 randomly assigned tracings and none reviewed tracings from his or her center. Local and central physician reviewers were instructed to categorize values as invalid if specified technical quality assurance standards were not met or the signal pattern suggested implausible values because of technical deficiencies. An intraclass correlation coefficient was calculated for continuous (numerical) variables and a kappa statistic was calculated for qualitative values with acceptable agreement defined a priori as an intraclass correlation coefficient of greater than 0.6. RESULTS: Filling cystometrogram baseline pressure, Valsalva leak point pressure, and volume and pressure measurements at maximum cystometric capacity had excellent intraclass correlation coefficients of 0.74 to 0.99. There were no significant differences between local and central physician reviewer means, indicating excellent agreement. CONCLUSIONS: With proper quality control measures in place and a set of standardized interpretive guidelines excellent interrater reliability between local and central physician reviewer can be achieved for numerical cystometrogram variables. PMID- 16697833 TI - Predictors of success for first stage neuromodulation: motor versus sensory response. AB - PURPOSE: We investigated whether intraoperative motor or sensory response is more predictive of successful sacral neuromodulation using the InterStim system. MATERIALS AND METHODS: A total of 35 patients with medically refractory frequency, urgency and urge incontinence were enrolled in the study. All patients underwent lead placement for quadripolar test stimulation under local anesthesia with intravenous sedation. Confirmation of correct lead placement was by observation of known motor and sensory responses that result from third sacral nerve stimulation. Motor and sensory responses were documented intraoperatively. Patients had a 1-week trial of stimulation, and those who had greater than 50% improvement in symptoms had placement of the implantable pulse generator. Those without at least 50% improvement in their symptoms had the quadripolar lead removed. RESULTS: Of the 35 patients enrolled 21 had successful quadripolar test stimulation and went on to permanent implantable pulse generator placement. Of the patients who had successful quadripolar test stimulation 95% demonstrated positive intraoperative motor response whereas only 21.4% of patients with unsuccessful quadripolar test stimulation demonstrated positive motor response. If only a positive sensory response was elicited, patients had only a 4.7% chance of having a positive quadripolar test stimulation. CONCLUSIONS: A positive quadripolar test stimulation (greater than 50% improvement in symptoms) with InterStim sacral neuromodulation is more likely when intraoperative lead placement results in positive motor response vs only sensory response. PMID- 16697835 TI - Failure of sacral nerve stimulation due to migration of tined lead. AB - PURPOSE: Stimulation of the sacral nerves is a commonly used treatment for frequency, urgency, urge incontinence, retention and other types of voiding dysfunction. Minimally invasive placement of a percutaneous permanent quadripolar tined lead into the sacral foramen has been described. No lead migration has been reported. We report on our experience with lead migration and the subsequent failure of InterStim in a large cohort of patients with a focus on possible diagnostic and salvage techniques. MATERIALS AND METHODS: Between February 2002 and April 2005 tined lead electrodes were implanted in the S3 foramen in 235 patients using the InterStim system. Patients with a good response during the testing phase (greater than 50% improvement) underwent placement of an implantable pulse generator. Position was confirmed by radiographic evaluation intraoperatively. Sacral radiographs were obtained at the first postoperative visit, after IPG placement and whenever there was a change in symptomatic response. RESULTS: There were 5 patients (2.1%) in whom treatment failed after a successful trial of stimulation due to lead migration. This was seen as early as 3 weeks and as late as 8 months. Migration of the lead occurred between first and second stage implantation in 1 of the 5 cases, and occurred after the second stage in 4 of 5. Anterior migration was noted in 4 patients and posterior migration was noted in 1. CONCLUSIONS: Lead migration after placement of the tined lead can occur and thus sacral radiographs should be routinely used. This complication can be easily resolved without significant morbidity to the patient. PMID- 16697836 TI - Outcomes following erosions of the artificial urinary sphincter. AB - PURPOSE: Artificial urinary sphincter urethral cuff erosion occurs in up to 5.0% of cases, presenting a complex management problem. We examine our experience with the eroded AUS, relating to preoperative risk factors, operative management and outcomes. MATERIALS AND METHODS: We reviewed the medical records of 637 patients undergoing bulbar urethral AUS implantation from 1990 to 2003 for demographic and surgical variables. RESULTS: Of the 637 records reviewed, 46 patients underwent 54 explantations of the AUS device for erosions, including 13 who had the primary implant performed at our institution and 33 being referred for management of erosion after implantation elsewhere. Our institution erosion rate was 2.2%. Mean followup after AUS reimplant following erosion was 27.8 months (range 1 to 180). Comorbidities were more prevalent in patients with erosions included hypertension (p = 0.006), coronary artery disease (p = 0.03), prior radiation therapy (p = 0.006) and prior AUS revisions (p = 0.0001). A majority of patients had persistent mild incontinence (0 to 1 pad daily in 29 or 56.8%), moderate (1 to 3 pads daily in 9 or 17.4%) and severe (more than 3 pads daily in 6 or 11.8%) incontinence after secondary AUS implantation. Patients who underwent reimplantation after AUS cuff erosions have a significantly higher rate of second erosions (in 16, 34.8%) within an average of 6.7 months (range 3 to 24), including our own 11.8% institutional rate (in 4). CONCLUSIONS: Our study suggests that patients with comorbidities including hypertension, coronary artery disease, prior radiation therapy and prior AUS revisions are more likely to have erosions of their AUS. Nevertheless, continence can still be salvaged using various strategies which optimize use of the remaining healthy urethral tissue. PMID- 16697838 TI - Inside out transobturator vaginal tape for the treatment of female stress urinary incontinence: interim results of a prospective study after a 1-year minimum followup. AB - PURPOSE: We analyzed the results of a prospective, observational trial designed to assess the safety and efficacy of the TVT-O procedure for female SUI. MATERIALS AND METHODS: Preoperative and postoperative evaluations included physical examination, and urinary symptom and quality of life scale questionnaires. RESULTS: Between March 2003 and December 2004, 253 patients with clinical and urodynamic diagnoses of SUI who fulfilled inclusion and exclusion criteria were enrolled in the trial and underwent the TVT-O procedure. No significant intraoperative complications were observed. One-year minimum followup was available on 99 of the initial 102 patients, of whom 16 had undergone concomitant pelvic organ prolapse surgical treatment. The SUI complete cure rate was 91%. No patient had vaginal or urethral erosion. Four patients required tape release or section. Frequency and urge symptoms improved after the operation (p <0.001). The severity of obstructive symptoms slightly increased postoperatively in the group of patients who did not undergo associated pelvic organ prolapse treatment (p <0.05), while maximum flow rates somewhat decreased (p <0.001) and post-void residual urine volumes somewhat increased (p <0.005). Most patients reported a significant decrease in incontinence severity and improvement in quality of life (p <0.0001). CONCLUSIONS: The results of this study, which suggest that the TVT-O procedure is a safe and efficient surgical treatment for female SUI, warrant further comparative evaluation of this procedure with retropubic and outside in transobturator approaches in appropriately designed, prospective, randomized trials. PMID- 16697839 TI - Treatment for unsuccessful tension-free vaginal tape operation by shortening pre implanted tape. AB - PURPOSE: We studied the efficacy of shortening the pre-implanted suburethral tape in patients with recurrent urodynamic stress incontinence after a TVT operation. MATERIALS AND METHODS: A total of 14 women, including 6 with ISD, were treated for recurrent urodynamic stress incontinence after the initial TVT operation by performing the shortening procedure under local anesthesia. Urodynamics, a 1-hour pad test, introital ultrasonography of the urethra and a cotton swab test were done before the procedure and 1 year postoperatively. RESULTS: All 14 patients completed the shortening procedure. Mean patient age was 47.2 years (range 43 to 66). Mean time between initial TVT and the shortening procedure was 4 months (range 3 to 14). Ten patients (71.4%) were objectively cured and treatment failed in 4 (2 with ISD and 2 with a fixed urethra). Mean operative time was 17 minutes (range 10 to 25). No intraoperative surgical complications were observed. The 1 hour pad test showed a decrease from a median of 9.0 gm to 1.0. Median postoperative hospital stay was 1 day (range 1 to 4). Spontaneous voiding with adequate post-void residual urine was noted in all patients before discharge home. CONCLUSIONS: Shortening a pre-implanted TVT tape for the treatment of recurrent urodynamic stress incontinence is a safe, effective and minimally invasive option requiring only a short hospital stay. However, ISD and an immobile urethra seem to be risk factors for failure. Long-term followup is needed to determine if this surgery achieves long-lasting results. PMID- 16697841 TI - Restoration of posterior aspect of rhabdosphincter shortens continence time after radical retropubic prostatectomy. AB - PURPOSE: Prolonged postoperative incontinence is a major drawback of RRP. Age, scars in the rhabdosphincter, nonnerve sparing surgery and postoperative sphincter insufficiency can cause temporary or definitive urinary incontinence. We believe that sphincter deficiency is the main cause of early incontinence. Urinary leakage results from the shortening of anatomical and functional sphincter length due to caudal retraction of the urethral sphincteric complex and disruption of the median posterior fibrous raphe. We describe a modification of the Walsh RRP that overcomes caudal retraction, reconstructs the posterior fibrous raphe and decreases time to continence. The primary study end point was early continence rate assessment. Long-term continence (1 year) and erectile function assessment were secondary end points. MATERIALS AND METHODS: To avoid caudal retraction of the urethrosphincteric complex, before completing the vesicourethral anastomosis the posterior semicircumference of the sphincter is joined to the residuum of Denonvilliers' fascia and fixed to the posterior bladder wall 1 to 2 cm cranial and dorsal to the new bladder neck. Vesicourethral anastomosis is subsequently performed with care taken not to involve the neurovascular bundles. A total of 161 patients with clinically confined disease underwent modified RRP (group 1). They were compared with a historical series of 50 patients who underwent standard RRP (group 2). Early continence was defined as no pad use but patients using 1 diaper were also considered continent. Continence, assessed prospectively as the number of pads daily, was evaluated 3, 30 and 90 days, and 1 year after catheter removal. The continence state was assessed by a multivariate logistic model. Erectile function was evaluated using the International Index of Erectile Function questionnaire preoperatively and after 18 months in patients younger than 65 years who underwent nerve sparing surgery. RESULTS: In group 1, 116 (72%), 127 (78.8%) and 139 patients (86.3%) were continent 3, 30 and 90 days after catheter removal compared with 7 (14%), 15 (30%) and 23 (46%), respectively, in group 2. One-year continence rates were 96% and 90%, respectively. Erectile function was similar in groups 1 and 2 (46% and 42%, respectively). Multivariate analysis showed that continence was significantly influenced by operation type, stage and patient age. CONCLUSIONS: Careful reconstruction of the posterior aspect of the rhabdosphincter markedly shortens time to continence. PMID- 16697842 TI - Nocturnal tumescence: a parameter for postoperative erectile integrity after nerve sparing radical prostatectomy. AB - PURPOSE: The exact process and time required for rehabilitation of erectile function after nerve sparing prostatectomy remain unclear to date. Different theories of the pathophysiology of postoperative erectile dysfunction are currently being discussed. In a prospective study we performed recordings of nocturnal penile tumescence and rigidity during the acute phase after nerve sparing radical prostatectomy, ie in the first night after removal of the catheter, to assess the organic penile integrity. MATERIALS AND METHODS: In 27 patients with local prostate carcinoma who had been sexually active before the intervention, we performed unilateral or bilateral nerve sparing radical prostatectomy. Preoperative sexual function of all patients was evaluated by the International Index of Erectile Function-5 questionnaire. On the day of catheter removal (postoperative day 7 to 14) an NPTR recording was performed on the following night with an erectometer (RigiScan). RESULTS: All patients had a preoperative IIEF score greater than 18. After removal of the catheter 25 of 27 patients (93%) showed 1 to 5 nocturnal rigidity increases by greater than 70% for at least 10 minutes. In a control group of 4 patients who underwent radical prostatectomy without nerve sparing, no nocturnal erections were recorded. CONCLUSIONS: NPTR recording during the acute phase after nerve sparing radical prostatectomy showed residual erectile function as early as the first night after catheter removal. These results are significant for selecting adequate pharmacological treatment for optimal therapy and rehabilitation of satisfactory erections and sexual function. In cases of early nocturnal tumescence, application of a PDE5 inhibitor can support successive organ rehabilitation. However, if tumescence does not occur, penile injection therapy is recommended. PMID- 16697843 TI - Synchronous prosthetic implantation through a transscrotal incision: an outcome analysis. AB - PURPOSE: The ease of the transscrotal approach for penile prosthesis implantation and the proximity of the urethra has allowed the evolution of a new strategy for dual implantation of an AUS and IPP at a single setting. Concerns regarding increased infection rates and poor outcomes have prevented widespread acceptance of this simultaneous implantation technique. A multi-institutional evaluation of dual prosthesis implantation was performed to determine the overall efficacy, safety and long-term success of this innovative approach. MATERIALS AND METHODS: We performed a multi-institutional, retrospective analysis in patients with 2 urological prostheses (AUS and IPP). Only patients undergoing dual implantation via a single transscrotal technique were included for evaluation. Comorbidities, infection rates, and intraoperative and postoperative complications were recorded. RESULTS: A total of 22 men underwent AUS and IPP device implantation between 2000 and 2003 in a synchronous manner. Mean followup was 17 months. Complications were urethral erosion in 2 patients (9%) and reservoir migration in 2 (9%), of whom 1 underwent revision. Importantly no patient experienced a prosthetic infection postoperatively. The overall revision rate was 14%, due to urethral erosion of the AUS in 2 patients and reservoir migration in 1. All patients reported urinary leakage requiring 1 pad daily or less. CONCLUSIONS: Our initial intermediate followup in patients who underwent synchronous dual prosthetic implantation was favorable. The inherent advantage of a single anesthetic event and a single transscrotal incision should encourage widespread acceptance of this technique. PMID- 16697844 TI - A novel microfluidic device for male subfertility screening. AB - PURPOSE: To our knowledge no simple, disposable and accurate test of semen quality currently exists. A novel technique to assess the motile sperm concentration of the human ejaculate has been designed and its results are presented. MATERIALS AND METHODS: In a micromachined device fluorescently labeled motile sperm traverse a hydrostatic microfluid line to a target detection cuvette. A microfluorometer assesses the fluorescence signal generated by sperm accumulating there throughout a 50-minute study period. A total of 21 semen specimens from men presenting to our university based reproductive endocrinology and infertility center were tested a total of 67 times. Semen parameters determined by computer assisted semen analysis were compared to the signal reported by the microfluidic device. RESULTS: The fluorescence signal increased throughout the data collection period for all samples. Pearson r values relating the device signal to total and progressive motile concentration were 0.79 and 0.80, respectively (each p <0.001). A signal threshold based on the aggregate data were established, correlating with the WHO standard of the normal total motile sperm concentration. As a screening test, the device was 94% sensitive and 97% specific for identifying samples with less than the WHO standard for the total motile concentration, and 96% sensitive and 90% specific when considering the progressive motile concentration. CONCLUSIONS: A novel microfluidic device is presented that enables accurate assessment of the motile sperm concentration in human ejaculate compared to computer assisted semen analysis. Its size and design demonstrate the feasibility of applying laboratory on chip technology to male infertility screening. PMID- 16697846 TI - The prostate cancer prevention trial: design, biases and interpretation of study results. AB - PURPOSE: We describe the complexities of the study design of the PCPT and how they influenced the end point chosen, trial implementation, analysis and interpretation of the results. MATERIALS AND METHODS: Data from the PCPT are provided to evaluate and quantify the potential biases of this trial design. RESULTS: Six potential sources of bias, including prostate specific antigen, digital rectal examination, prostate biopsy technique, study medication nonadherence and contamination, and transurethral prostate resection are presented. These biases resulted in the need for the end of study biopsy to evaluate the trial objectives. CONCLUSIONS: There were a large number of known and potential biases that worked for and against finasteride. Because of the trial design and inherent biases, it is imperative that interim biopsy results should be interpreted with caution. While the period prevalence end point that relied on an end of study biopsy was perhaps not the most clinically relevant, it was the only way to remove as much bias as possible and meet the study objective of determining if finasteride could decrease the risk of prostate cancer. The success of the PCPT depended on constant scrutiny by the Data and Safety Monitoring Committee to monitor these biases. The design and biopsy assumptions outlined at the inception of the trial were met, including adherence and contamination rates, the for-cause biopsy rate and the final percent of men with study end points. PMID- 16697847 TI - The continued decline of formal urological education of medical students in the United States: does it matter? AB - PURPOSE: Survey analyses of 50 years document a substantial decline in the formal urological education of United States medical students, resulting in repeated calls by urologists to reverse this trend. Has this trend continued during the last decade? If so, should we be concerned? We performed a national survey to provide an updated assessment of urological education at United States medical schools. MATERIALS AND METHODS: All 321 applicants to the 2003 to 2004 United States urology residency match and 527 applicants to a Boston area emergency medicine residency program in 2003 to 2004 from accredited United States medical schools were invited to participate in the study. On a validated, anonymous online survey, these students were asked 1) did their school offer a required and/or elective clinical rotation in urology, and 2) if so, what was its duration and in which year was it usually completed. RESULTS: Curricular data were obtained on 110 of 125 (88%) accredited United States medical schools. Of United States schools 17% (17.3%, SE 1.2%, 19 of 110) have required clinical rotations in urology; these required rotations are 1 to 2 weeks in duration and, in 79% of cases, conducted in year 3 of medical school. Those medical schools that require a clinical rotation in urology do not have a greater percentage of their students entering the urology match (Mann-Whitney U test p = 0.540). CONCLUSIONS: Formal urological education of United States medical students has continued to erode during the last decade, but the impact of this trend on student learning and career choice is not clear. Further research is required to determine whether these changes are of concern for the quality of applicants to urological training and for the future of urological care in the United States. PMID- 16697849 TI - Validity and reliability of parental report of frequency, severity and risk factors of urinary tract infection and urinary incontinence in children. AB - PURPOSE: We describe the validity and reliability of a questionnaire designed to determine frequency, severity and risk factors of urinary tract infection and daytime urinary incontinence in primary school-age children. MATERIALS AND METHODS: Based on published validated questionnaires and advice from content experts, a questionnaire was developed and piloted in children attending outpatient clinics. Construct validity for parent report of frequency and severity of daytime urinary incontinence was tested by comparison with a daily accident diary in 52 primary school children, and criterion validity of parent report for UTI was verified by comparison with the reference standard (urine culture) in 100 primary school children. Test-retest reliability of the questionnaire was assessed in 106 children from primary schools. RESULTS: There was excellent agreement between the questionnaire and accident diary in severity (weighted kappa 0.94, 95% confidence intervals 0.85 to 1.03) and frequency of daytime urinary incontinence (0.88, 0.7 to 1.0). Parents reported urinary tract infection in 15% of children, compared to a positive urine culture in 8% (sensitivity 100% and specificity 68.5%). Test-retest reliability of the questionnaire was excellent (mean k 0.78, range 0.61 to 1.00). CONCLUSIONS: Parents overreport UTI by about 2-fold but can recall frequency and severity of daytime urinary incontinence well during a 3-month period. The developed questionnaire is a valid tool to estimate frequency, severity and risk factors of daytime urinary incontinence and UTI in primary school children. PMID- 16697850 TI - Home uroflowmetry biofeedback in behavioral training for dysfunctional voiding in school-age children: a randomized controlled study. AB - PURPOSE: We studied the added value of home uroflowmetry for biofeedback training compared to added attention and standard therapy in a multicomponent behavioral training program for voiding disorders in school-age children. Little is known about the role of biofeedback by home uroflowmetry for dysfunctional voiding due to NNBSD in children. MATERIALS AND METHODS: A randomized controlled study was conducted at an outpatient pediatric incontinence university clinic from January 2000 to June 2003. A total of 192 children 6 to 16 years old who were suffering from recurrent urinary tract infections with or without urge incontinence were screened for NNBSD. Of 143 eligible patients 44 were randomly allocated to receive 8 weeks of standard therapy (outpatient behavioral therapy), 46 to receive 8 weeks of home video instructions together with standard therapy and 53 to receive 8 weeks of home uroflowmetry biofeedback together with standard therapy. After 8 weeks all treatment groups proceeded with standard therapy for 16 weeks, after which prophylaxis with antibiotics was stopped and patients were followed for another 6 months. Main outcome measurement was total relief of complaints, namely urinary tract infections and, if present, incontinence, at 12 months after randomization. RESULTS: At baseline there was no reason to predict major incomparabilities between the groups. In an intent to treat analysis there was no difference in total relief between standard treatment (44%) and added video instruction (42%, RR 0.96, 95% CI 0.59 to 1.56). Total relief in the added home uroflowmetry group (55%) was higher than with standard therapy (RR 1.24, 95% CI 0.80 to 1.93), although the difference was not statistically significant. A per protocol analysis suggested that the groups with added home uroflowmetry showed better total relief than the pooled groups with standard therapy and those with added video (RR 1.40, 95% CI 0.98 to 2.00). CONCLUSIONS: Home uroflowmetry appears to be a useful adjunctive treatment for the reduction of complaints in children with dysfunctional voiding due to nonneurogenic bladder-sphincter dyssynergia. PMID- 16697852 TI - Comparison of laparoscopic and open partial nephrectomy for duplication anomalies in children. AB - PURPOSE: We compared the outcome of laparoscopic vs open PN for duplication anomalies at our institution. MATERIALS AND METHODS: We retrospectively reviewed the records of all patients undergoing PN within the last 4 years. RESULTS: A total of 34 patients (16 females) were divided into 2 groups. Group 1 consisted of 20 patients undergoing open PN between 2000 and 2003, and group 2 consisted of 14 patients undergoing laparoscopic PN between 2003 and 2004. Mean patient age was 21 months in group 1 and 18 months in group 2. Diagnosis was ectopic ureter in 18 patients, ureterocele in 11, VUR in 4 and ureteropelvic junction obstruction in 1. Cystoscopy was performed as part of the procedure in 30% of the patients in group 1 and 100% of those in group 2. Simultaneous lower tract procedures were performed in 3 patients in group 1 and 2 patients in group 2. Mean duration of PN was 115 minutes for group 1 and 180 minutes for group 2. There was no significant bleeding or need for transfusion except in 1 patient in the open group. Median hospitalization was 3 days for group 1 and 2 days for group 2. Mean analgesic requirement was 2.3 doses of opioids and 2 doses of ketorolac for group 1, and 3.2 doses of opioids for group 2. Acetaminophen only was used in 3 of 20 patients in group 1 and 5 of 14 in group 2. There were 2 complications in each group, namely 1 case of ureteral bleeding and 1 lower pole ureteral injury in group 1, and 1 omental hernia and 1 urinoma in group 2. CONCLUSIONS: Laparoscopic PN is feasible even in small infants, and the results are comparable to the open procedure. Length of hospitalization was shorter in the laparoscopic group. In our series the learning curve for this technique was rapid, and after a few cases the procedure could be done in the same time as open surgery, with the advantages offered by laparoscopy. PMID- 16697853 TI - Laparoscopic renal denervation and nephropexy for autosomal dominant polycystic kidney disease related pain in adolescents. AB - PURPOSE: We present our initial experience with laparoscopic renal denervation and nephropexy for ADPKD related pain in the adolescent population. MATERIALS AND METHODS: Four patients 15 to 19 years old previously diagnosed with ADPKD presented with chronic pain refractory to narcotic analgesics. These 4 patients underwent laparoscopic renal denervation of 5 kidneys. RESULTS: Mean operative time was 136 minutes and mean hospital stay was 2.75 days. All patients were pain free at discharge home and remained pain-free at a mean followup of 11.5 months. CONCLUSIONS: We believe that laparoscopic renal denervation plus nephropexy is a promising option for uncontrolled ADPKD related pain in the adolescent population. PMID- 16697855 TI - Outcome analysis of retroperitoneal laparoscopic heminephrectomy in children. AB - PURPOSE: Retroperitoneal laparoscopic heminephrectomy is a relatively new, well tolerated, minimally invasive alternative to open surgery for children with a nonfunctioning moiety in a duplex kidney. We collected data to assess outcomes in our experience with this procedure. MATERIALS AND METHODS: Data were retrospectively gathered on all patients operated on for retroperitoneal laparoscopic heminephrectomy at our institution between February 2001 and November 2004. Followup ultrasound was performed in all patients, and additional imaging was done only when clinically indicated. RESULTS: Retroperitoneal laparoscopic heminephrectomy was performed in 18 girls and 4 boys (18 upper and 5 lower pole moieties). Indications included ureterocele (12 patients), ectopic ureter (9) and vesicoureteral reflux (5). Mean age at surgery was 5 years (range 4 months to 18 years). Four patients required conversion to an open procedure due to inadequate exposure and were excluded from further review. Urine leaks developed postoperatively in 3 patients and were treated with Foley catheter drainage and/or stent placement. Two patients were noted to have large (more than 5 cm) asymptomatic retroperitoneal fluid collections on followup ultrasound. Functional loss of the remaining ipsilateral moiety was confirmed by DMSA scan in 2 children (ages 6 and 7 months). Doppler studies confirmed normal hilar blood flow in both cases. Clinical presentation for this subgroup included fever on postoperative day 3 in 1 patient and hypertension at 32 months postoperatively in 1. Mean followup was 33 months (range 3 to 56). CONCLUSIONS: Although retroperitoneal laparoscopic renal ablative surgery in children appears to be relatively safe and is associated with low morbidity, serious complications may occur with subtle or even late presentation. PMID- 16697858 TI - Sexual function following bowel vaginoplasty. AB - PURPOSE: We review our 23-year experience with bowel vaginoplasty, with particular attention to postoperative quality of life and sexual function. MATERIALS AND METHODS: We reviewed the records of 57 patients who underwent bowel replacement vaginoplasty between 1980 and 2004. A total of 42 patients had the Mayer-Rokitansky syndrome, of whom 6 had varying forms of intersexuality, 6 had undergone surgery for pelvic malignancy, 1 had aphallia, 1 had cloacal exstrophy and 1 was the survivor of a conjoined twin separation. Replacement vaginoplasty was done using sigmoid colon in 39 patients, ileum in 9 and cecum in 9. Followup ranged from 18 months to 24 years, with a mean of 8.8 years. Outcome was evaluated by retrospective chart review, and the FSDQ, a validated, IRB approved instrument, was used to evaluate postoperative sexual function. RESULTS: Among the 57 patients postoperative sexual function was evaluated in 44, 9 were lost to followup and 4 were considered too young for evaluation. Of the 44 patients 36 responded to the FSDQ, 6 refused and 2 were unable to complete the questionnaire adequately. Of the 36 patients who responded 15 were married and 31 were sexually active. On a scale of 0 to 5, 28 patients (78%) reported sexual desire, 33% sexual arousal, 33% sexual confidence and 28 (78%) sexual satisfaction. In addition, 20 patients (56%) reported frequent orgasms, 8 (22%) occasional orgasms and 8 (22%) no orgasms. A total of 32 patients (89%) reported adequate lubrication for intercourse and 2 reported dyspareunia. Two of the 36 patients performed home dilation and required estrogen suppositories. A total of 34 patients used home douching and 20 required pads for mucus production. CONCLUSIONS: It appears that isolated bowel segments provided excellent tissue for vaginal replacement. For technical reasons we believe that colonic segments, particularly sigmoid, are preferable to small bowel. Sexual function following bowel vaginoplasty appears to be adequate and durable. PMID- 16697859 TI - Torsion of the cryptorchid testis--can it be salvaged? AB - PURPOSE: The literature concerning undescended testis mainly concentrates on the increased risks of infertility and development of germ cell tumors. Yet the UDT also appears to be at higher risk for torsion compared to the normally descended testis, and this issue is relatively poorly addressed. We reviewed all cases of torsion of UDTs operated on at our hospital during the last 20 years in an attempt to characterize better this condition and its salvageability. MATERIALS AND METHODS: In this retrospective clinical study we reviewed and analyzed all cases of testicular torsion involving UDT operated on at our hospital between 1984 and 2004. RESULTS: A total of 11 children were operated on at our hospital for torsion of undescended testis between 1984 and 2004. Patient age ranged from 1 month to 18 years (median 7.5 months). In all cases unilateral torsion of undescended testis was diagnosed, with 73% of cases involving the left side. Clinical symptoms included inguinal swelling and erythema associated with a tender, firm mass palpated in the groin area and an empty ipsilateral hemiscrotum. Doppler ultrasound examination was routinely performed in the last 7 patients to confirm the diagnosis. During inguinoscrotal exploration severe ischemia or overt necrosis of the testis was found in 5 of 11 cases. Three of these 5 cases were managed by orchiectomy, while in the other 2 cases the testis subsequently vanished. In the 6 patients who exhibited some improvement following detorsion and warming of the tissue the testes were preserved and orchiopexy was performed. Followup was available in only 5 patients, with vanishing of the torsed testis observed in 4 and a normal testicle detected 21 years postoperatively in 1 patient who was diagnosed early. CONCLUSIONS: This series clearly demonstrates poor rates of surgical salvage, which we mainly attribute to delays in parental response and in primary physician referral to the hospital. Parents, who have a pivotal role in early diagnosis, were usually unaware of this urological emergency, and some were surprisingly unaware of the presence of cryptorchidism. By increasing the awareness regarding this entity among members of the medical community and parents, we hope that torsion of the cryptorchid testis (literally, "hidden testis") will no longer necessarily be synonymous with "crypt-torsion" ("hidden torsion"). PMID- 16697861 TI - Assessment of serum follicle-stimulating hormone level and testicular volume for prediction of paternity potential in pubertal boys who underwent bilateral orchiopexy in childhood. AB - PURPOSE: We examined infertile adults for the presence of spermatozoa in the testes, serum FSH level and testicular volume to predict paternity potential in pubertal boys who had undergone bilateral orchiopexy in childhood. MATERIALS AND METHODS: The study included 58 idiopathic infertile men (group 1), 14 infertile men who had undergone bilateral orchiopexy in childhood (group 2) and 21 pubertal boys who had undergone bilateral orchiopexy in childhood (group 3). We evaluated the correlation between FSH level and testicular volume in all groups. In groups 1 and 2 the testes were examined for the presence of spermatozoa using bilateral testicular biopsy samples. RESULTS: Judging from the presence of spermatozoa in the testes and correlation between FSH level and testicular volume, 14 patients in group 1 (24%) were categorized as being at high risk, 32 (55%) at intermediate risk and 12 (21%) at low risk for loss of paternity potential, with spermatozoa positive rates of 0%, 49% and 100%, respectively. In comparison, 4 patients in group 2 (29%) were categorized as being at high risk, 7 (50%) at intermediate risk and 3 (21%) at low risk, with spermatozoa positive rates of 0%, 43% and 100%, respectively. According to this categorization, 19% of pubertal boys in group 3 were at high risk and may already have lost their paternity potential. CONCLUSIONS: Despite bilateral orchiopexy in childhood, some male adolescents may not have the potential for paternity. PMID- 16697862 TI - Gonadoblastoma. PMID- 16697863 TI - Classification of renal neoplasms based on molecular signatures. AB - PURPOSE: Gene expression microarray studies have demonstrated distinct molecular signatures for different types of renal neoplasms based on overall gene expression patterns. However, in most of these studies the investigators used renal tumors with defined histology. We analyzed a test set of renal tumors in double-blind fashion using recently established molecular profiles of renal tumors as benchmarks. MATERIALS AND METHODS: A total of 16 consecutive nephrectomies performed for neoplasms at a single urological service were subjected to gene expression profiling using cDNA chips containing 21,632 genes. Analysis was clustered with our previously established molecular profiles of 91 histologically defined kidney neoplasms and comparative genomic microarray analysis while blinded to tumor histology and clinical information. RESULTS: With molecular analysis 9, 4, 2 and 1 tumors were classified as clear cell, papillary RCC, chromophobe RCC, and renal oncocytoma, respectively. Histopathological evaluation was concordant in 14 tumors. One of the 2 tumors with a discrepancy between molecular and pathological diagnoses was composed of oncocytoma and high grade clear cell RCC, and the other was chromophobe RCC that histologically mimicked papillary RCC. CONCLUSIONS: We report the feasibility of the molecular diagnosis and classification of unknown renal neoplasms. Molecular diagnosis appears to be reliable and comparable to the standard of urological pathology. This molecular method may be a potentially useful test for establishing an accurate diagnosis that can impact clinical management. PMID- 16697864 TI - Acute integrity of closure for partial nephrectomy: comparison of 7 agents in a hypertensive porcine model. AB - PURPOSE: We assessed the acute effectiveness of closure after partial nephrectomy of 7 techniques in a large hypertensive porcine model using shallow and deep resections to approximate clinical situations. MATERIALS AND METHODS: Open surgical partial nephrectomy with hilar clamping was performed in pigs weighing 150 to 200 lbs, including small-a quarter length and a quarter width of kidney, medium-a third length and a third width of kidney, and into the renal sinus and up to the collecting system, and large-lower pole heminephrectomy at the renal sinus. Seven agents were compared after a single application, namely thrombin/collagen granules, polyethylene glycol hydrogel, fibrin glue, thrombin/gelatin granules, cyanoacrylate glue, fibrin glue/gelatin sponge and sutured bolster. Failure and success were determined by the presence or absence of bleeding, respectively, after unclamping and by an increase in SBP to 100 and then to 200 mm Hg with dopamine infusion. RESULTS: Of 70 partial nephrectomies the success rates were 33% and 14% for thrombin/collagen granules, and 67% and 0% for polyethylene glycol hydrogel in small and medium resections; 100%, 71% and 0% for fibrin glue, and 100%, 86% and 0% for thrombin/gelatin granules in small, medium and large resections; and 67% and 80% for cyanoacrylate glue, 100% and 20% for fibrin glue/gelatin sponge, and 100% for sutured bolster in medium and large resections, respectively. Of the kidneys that did not bleed at an SBP of 100 mm Hg 31% bled at 200 mm Hg. CONCLUSIONS: There is considerable variability among agents. Most were effective for small resections and some worked for medium resections but for large resections only sutured bolster was consistently effective. SBP also appears to be an important factor. These results bear on the selection of techniques during laparoscopic partial nephrectomy. PMID- 16697865 TI - Evaluation of survivin reverse transcriptase-polymerase chain reaction for noninvasive detection of bladder cancer. AB - PURPOSE: Survivin, a member of the inhibitor of apoptosis gene family, is expressed in most common cancers. We investigated the expression pattern of survivin in the tumors of patients with bladder cancer and assessed the diagnostic potential of RT-PCR detection of survivin mRNA in urine. MATERIALS AND METHODS: RT-PCR was used to analyze mRNA expression of survivin in 161 cases of bladder cancer, including TCC in 97, SCC in 53 and adenocarcinoma in 11, and their matched nontumor tissues. Urine specimens (50 ml) were collected from 84 patients in whom bladder cancer was documented by transurethral resection or biopsy, 41 with nonbladder cancer urological diseases and 42 healthy volunteers. Total RNA was extracted from urine sediments and RT-PCR was performed for survivin. RESULTS: Survivin expression was detected in all bladder cancer tissues. In contrast, survivin was not detectable in normal urothelium specimens. Urinary survivin was detected in urine samples from 51 of 53 patients with TCC, 22 of 25 with SCC and 6 of 6 with adenocarcinoma. Overall sensitivity was 94%. Survivin mRNA was not detected in any healthy volunteers. Positive results were obtained in 2 patients with renal cell carcinoma, 1 with hematuria and 1 with a contracted bladder but in none with other urological diseases. Overall specificity was 95%. CONCLUSIONS: Survivin mRNA detection in urine sediment using RT-PCR shows high sensitivity and specificity for bladder cancer. It may prove useful for the routine screening and monitoring of patients. PMID- 16697866 TI - Intraperitoneal chemotherapy for the prevention of transitional cell carcinoma implantation. AB - PURPOSE: Tumor spillage from bladder perforation during transurethral resection of a bladder tumor or during cystectomy risks seeding the peritoneum with TCC. Current therapy is irrigation with sterile water with an unknown extent of clinical benefit. Intraperitoneal chemotherapy for other human cancers has demonstrable benefit but to our knowledge it has never been investigated for TCC. We investigated whether intraperitoneal chemotherapy can prevent TCC implantation in a murine model of tumor spillage and whether water irrigation is beneficial. MATERIALS AND METHODS: Laparotomy was performed in 28 Fischer 344 rats (National Cancer Institute, Frederick, Maryland) to instill 1 x 10 AY-27 TCC cells. Mitomycin (10 mg/m) was instilled in 9 rats and saline was used in the control group. A third group underwent lavage with sterile water. At sacrifice after 2 weeks tumors were measured in mm and weighed. A followup experiment of 4-week survival used 5 mg/m mitomycin and added a fourth group treated with water lavage plus mitomycin. RESULTS: All 9 rats in the saline control group had gross tumors at the laparotomy site as well as gross carcinomatosis. The 10 water lavage rats also demonstrated gross tumors but of smaller size (p = 0.02). All rats treated with mitomycin had no gross or microscopic evidence of tumor growth anywhere in the peritoneum. In experiment 2 none of the rats treated with lower dose mitomycin had gross or microscopic tumors regardless of water lavage. CONCLUSIONS: Intraperitoneal chemotherapy prevents TCC implantation in a murine model of tumor spillage. Water lavage decreases the tumor burden but it cannot effectively sterilize the peritoneum of tumor. PMID- 16697867 TI - Multifocal urothelial cancers with the mutator phenotype are of monoclonal origin and require panurothelial treatment for tumor clearance. AB - PURPOSE: UC is a disease of the entire urothelium, characterized by multiplicity and multifocality. The clonal relationship among multiple UCs has implications regarding adjuvant chemotherapy. It has been investigated in studies of chromosomal alteration and single gene mutation. However, these genetic changes can occur in unrelated tumors under similar carcinogenic selection pressures. Tumors with high MSI have numerous DNA mutations, of which many provide no selection benefit. While these tumors represent an ideal model for studying UC clonality, their low frequency has prevented their previous investigation. MATERIALS AND METHODS: We investigated 32 upper and lower urinary tract UCs with high MSI and 4 nonUC primary cancers in 9 patients. We used the high frequency and specificity of individual DNA mutations in these tumors (MSI at 17 loci) and the early timing of epigenetic events (methylation of 7 gene promoters) to investigate tumor clonality. RESULTS: Molecular alterations varied among tumors from different primary organs but they appeared related in the UCs of all 9 patients. While 7 patients had a high degree of concordance among UCs, in 2 the UCs shared only a few similar alterations. Genetic and epigenetic abnormalities were frequently found in normal urothelial samples. CONCLUSIONS: Multiple UCs in each patient appeared to arise from a single clone. The molecular order of tumor development varied from the timing of clinical presentation and suggested that residual malignant cells persist in the urinary tract despite apparent curative surgery. These cells lead to subsequent tumor relapse and new methods are required to detect and eradicate them. PMID- 16697868 TI - Triclosan loaded ureteral stents decrease proteus mirabilis 296 infection in a rabbit urinary tract infection model. AB - PURPOSE: Infection and encrustation remain major limitations of ureteral stent use and to our knowledge no device has completely overcome these obstacles to date. Triclosan is a biocide currently used in a plethora of consumer and medical products that has recently been loaded into a ureteral stent. Using a rabbit model of UTI we examined the effects of triclosan impregnated stent segments on the growth and survival of Proteus mirabilis, a uropathogen commonly associated with device related UTI and encrustation. MATERIALS AND METHODS: A total of 48 male New Zealand White rabbits were instilled transurethrally with 1 x 10(6) P. mirabilis 296. A stent curl from a triclosan eluting, Percuflex Plus or Optima ureteral stent was placed intravesically. Urine was cultured on days 1, 3 and 7. On day 7 the stents were assessed for encrustation and viable organisms, while the bladders were scored for the degree of inflammation. RESULTS: Throughout the study urine isolated from the triclosan group contained significantly fewer viable organisms than controls with 7 of 13 animals completely clearing the infection by day 7. Similarly 9 of 13 triclosan eluting stents showed no viable organisms upon recovery and the remaining 4 showed significantly fewer organisms than controls. Urine and stents in all controls were positive for P. mirabilis at all time points. Although there was no significant difference in encrustation among the groups, bladders harvested from the triclosan group demonstrated significantly less inflammation. CONCLUSIONS: Triclosan eluting stents greatly decreased P. mirabilis growth and survival in a rabbit UTI model compared to controls. These stents may prove useful for decreasing device related P. mirabilis UTIs. PMID- 16697869 TI - Establishing composition and structure of intact urinary calculi by x-ray coherent scatter for clinical laboratory investigations. AB - PURPOSE: Current urinary stone analysis techniques are limited in their abilities to simultaneously characterize composition and structure. Laboratory techniques such as IRS and x-ray diffractometry require small powdered samples for analysis, rendering composition results dependent on the choice of sample and its preparation. We investigated the application of x-ray CS analysis to identify topographic urinary stone composition ex vivo. CS is essentially a transmission based x-ray diffractometry method that depends on molecular structure and, therefore, can distinguish different compounds. Diagnostic x-ray equipment facilitates the examination of structural arrangements of minerals in intact calculi. MATERIALS AND METHODS: Tomographic images of intact calculi CS properties were acquired with a purposely built scanner. Composition is extracted from CS by fitting a library of common pure stone component CS signatures to those of the unknown sample in each image pixel. Two zones per stone were isolated (powdered) for IRS composition analysis for comparison with CS maps at these locations. Each stone was also independently analyzed for bulk composition by IRS analysis. RESULTS: CS composition maps revealed the spatial arrangement of minerals in intact calculi. IRS results showed good agreement with CS in the selected regions of interest. Bulk composition by IRS was noted to miss some important stone components, indicating that the choice of sample may skew composition results. CONCLUSIONS: CS from diagnostic x-rays can be used to identify structure and composition in mixed urinary calculi nondestructively. The tissue specific CS images presented support the development of CS analysis as a means of identifying stone composition characteristics in the laboratory. PMID- 16697870 TI - Botulinum-A toxin injections into the detrusor muscle decrease nerve growth factor bladder tissue levels in patients with neurogenic detrusor overactivity. AB - PURPOSE: We investigated the effects of BTX-A on visceral afferent nerve transmission by measuring bladder tissue NGF levels in patients with neurogenic detrusor overactivity before and after intravesical treatment with BTX-A. We also compared the bladder tissue NGF content with clinical and urodynamic data. MATERIALS AND METHODS: A total of 23 patients underwent clinical evaluation and urodynamics with detection of the UDC threshold, maximum pressure and maximum cystometric capacity before, and at the 1 and 3-month followups. Endoscopic bladder wall biopsies were also obtained at the same time points. NGF levels were measured in tissue homogenate by enzyme-linked immunosorbent assay (Promega, Madison, Wisconsin). RESULTS: At 1 and 3 months mean catheterization and incontinent episodes were significantly decreased (p <0.05 and <0.001, respectively). On urodynamics we detected a significant increase in the UDC threshold and maximum cystometric capacity, and a significant decrease in UDC maximum pressure at the 1 and 3-month follow-ups compared to baseline (each p <0.001). At the same time points we detected a significant decrease in NGF bladder tissue content (each p <0.02). CONCLUSIONS: BTX-A intravesical treatment induces a state of NGF deprivation in bladder tissue that persists at least up to 3 months. As caused by BTX-A, the decrease in acetylcholine release at the presynaptic level may induce a decrease in detrusor contractility and in NGF production by the detrusor muscle. Alternatively BTX-A can decrease the bladder level of neurotransmitters that normally modulate NGF production and release. PMID- 16697872 TI - Sildenafil citrate inhibits agonist induced contractions in isolated rat seminal vesicles. AB - PURPOSE: Sildenafil is reported to regulate smooth muscle contractility through nitric oxide-cyclic guanosine monophosphate, not only in the corpus cavernosum. Its possible effects on seminal vesicle contractility might be of importance with respect to premature ejaculation. We investigated the effects of sildenafil citrate (Pfizer, New York, New York) on agonist induced isometric contractions of the rat seminal vesicle in vitro. MATERIALS AND METHODS: Seminal vesicles isolated from adult male Wistar rats were suspended in an organ bath and contracted by NE (10 microM), ACh (10 microM) or KCl (60 mM) (Sigma, Deisenhofen, Germany). The effects of sildenafil citrate (100 to 300 microM) were evaluated in terms of mean contraction amplitude, the area under force-time curves and isometric contractility indexes. RESULTS: Sildenafil citrate (300 microM) significantly inhibited the mean amplitude +/- SEM of contractile responses induced by NE (1,061 +/- 153 vs 271 +/- 65 mg, p <0.0001), ACh (475 +/- 51 vs 68 +/- 17 mg, p <0.0001) and KCl (546 +/- 71 vs 59 +/- 18 mg, p <0.0002). It also caused dose dependent concomitant decreases in the area under force-time curves. Additionally, pretreatment with sildenafil citrate markedly prevented the contractile response to NE, ACh and KCl. CONCLUSIONS: Our results indicate that sildenafil citrate inhibits the contractions of isolated rat seminal vesicle that are induced by NE, ACh or KCl. Future studies may support an in vivo effect of sildenafil for delaying or inhibiting seminal vesicle emission, thereby, promoting improvement in patients with premature ejaculation. PMID- 16697871 TI - Effect of doxazosin with and without rho-kinase inhibitor on human corpus cavernosum smooth muscle in the presence of bladder outlet obstruction. AB - PURPOSE: We investigated the relationship of adrenergic responses in corpus cavernosum tissues in the presence of BOO using the alpha1-adrenergic receptor antagonist doxazosin (Pfizer, New York, New York) and the rho-kinase inhibitor Y 27632 (Calbiochem, San Diego, California). MATERIALS AND METHODS: CCSM tissue was obtained from patients who underwent penile prosthesis implantation. Patients were divided into 2 groups according to the presence of BOO. The submaximal (EC80) concentration of phenylephrine (Sigma Chemical Co., St. Louis, Missouri) was calculated by evaluating adrenergic activity responses with cumulatively applied phenylephrine. After achieving a stable contraction plateau test compounds were put in an organ bath. The relaxant potencies of doxazosin and Y 27632 were expressed as the percent of inhibition of the contraction plateau induced EC80 concentration of phenylephrine. Relaxation responses in the 2 groups were compared. RESULTS: At the highest dose of increasing concentrations phenylephrine generated 70% more contraction response in the BOO positive group than in the BOO negative group. Doxazosin and Y-27632 caused concentration dependent relaxation in CCSM precontracted by phenylephrine. With doxazosin significantly higher relaxation responses were attained in the BOO positive group in terms of log IC50 and the maximal relaxation response (p = 0.0353 and 0.0003, respectively). Maximum relaxation responses following Y-27632 administration were significantly higher in the BOO positive group. CONCLUSIONS: The contractility of human corpus cavernosum is increased in the presence of BOO. Doxazosin and Y 27632 generate effective CCSM relaxation in the presence of BOO. Doxazosin and Y 27632 may be the alternatives for the treatment of erectile dysfunction associated with BPH. PMID- 16697874 TI - Re: reproducibility of lymphoscintigraphy for lymphatic mapping in patients with penile carcinoma. PMID- 16697875 TI - Re: treatment delay and prognosis in invasive bladder cancer. PMID- 16697876 TI - Re: treatment of chronic bacterial prostatitis with levofloxacin and ciprofloxacin lowers serum prostate specific antigen. PMID- 16697878 TI - Re: hypogonadism and metabolic syndrome: implications for testosterone therapy. PMID- 16697881 TI - Re: potency following robotic radical prostatectomy: a questionnaire based analysis of outcomes after conventional nerve sparing and prostatic fascia sparing techniques. PMID- 16697882 TI - Re: what makes a medical student avoid or enter a career in urology? Results of an international survey. PMID- 16697884 TI - Letter from the guest editor: there but for the grace of ... PMID- 16697885 TI - Letter from the editor: the role of the radiologist in oncologic imaging. PMID- 16697886 TI - Case of the season: gossypiboma. PMID- 16697887 TI - Role of image-guided percutaneous needle biopsy in cancer staging. PMID- 16697888 TI - Breast cancer staging. PMID- 16697889 TI - Imaging upper gastrointestinal malignancy. PMID- 16697890 TI - Imaging colorectal cancer. PMID- 16697891 TI - Radiologic diagnosis and staging of renal and bladder cancer. PMID- 16697892 TI - Prostate cancer imaging. PMID- 16697894 TI - Finding protein kinase hits using structural information. PMID- 16697895 TI - Blunting the Swiss army knife of hepatitis C virus: inhibitors of NS3/4A protease. PMID- 16697896 TI - Peptide deformylase inhibitors. PMID- 16697897 TI - Clinically useful vanilloid receptor TRPV1 antagonists: just around the corner (or too early to tell)? PMID- 16697898 TI - Recent medicinal chemistry of the histamine H3 receptor. PMID- 16697899 TI - Recent progress in cannabinoid research. PMID- 16697900 TI - Oxytocin antagonists as potential therapeutic agents for the treatment of preterm labour. PMID- 16697902 TI - Minimally invasive surgery for valve disease. AB - Heart valve surgery evolved since the early 1960s toward routine clinical application with good patient outcome. Different surgical techniques and valve prostheses have been developed. Thus standard procedures were continuously established. The different surgical procedures have now gained widespread clinical acceptance with good patient outcomes. Aortic valve stenosis and mitral valve incompetence are the most frequently acquired heart valve lesions in the western communities. Usually such lesions reach clinical significance in patients during their fifth to eighth decade of life. Standard surgical techniques of aortic valve repair and mitral valve replacement or repair result in persistent cure of the disease. Surgical access was gained using conventional lateral thoracotomies in the early days and later on using median sternotomy. Minimally invasive techniques, mostly by a partial sternotomy for the aortic and a lateral minithoracotomy for the mitral position, have been increasingly applied to improve patient outcome since the mid 1990s. At specialized centers these techniques have evolved as clinical standard allowing all different valve procedures to be safely performed. Patient recovery is fast leading to a significant improvement in the individual's quality of life. Minimally invasive valve surgery can be considered the standard approach and will reach more widespread clinical application. PMID- 16697903 TI - Characterization of monoclonal antibodies directed against the canine distemper virus nucleocapsid protein. AB - We have established four monoclonal antibodies (MAbs) against the nucleocapsid protein (NP) of canine distemper virus (CDV). A competitive binding assay has revealed that the MAbs are directed against two antigenic domains. An immunofluorescence assay using a series of deletion clones of the NP and an immunoprecipitation assay using the NP have revealed that two of the MAbs recognize the C-terminal region of the NP while the other two recognize the tertiary structure of the N-terminal domain. These MAbs reacted with all eight strains of CDV used in this study, but showed different reactivities against measles virus and rinderpest virus. PMID- 16697905 TI - Paula Clayton: her role in the Development of the Field of Affectiveology. PMID- 16697904 TI - Serological evidence for Japanese encephalitis and West Nile viruses in domestic animals of Nepal. AB - A regional survey was conducted in Nepal for antibody to Japanese encephalitis virus (JEV) in domestic animals. Sera from pigs, and limited numbers of ducks and horses were collected from 16 districts in 2002-2003 and subjected to three serological tests. Of 270 porcine sera tested by C-ELISA, 55% were found positive for the presence of antibodies against Japanese encephalitis virus. Additional testing for IgM antibody to JEV revealed less than 2% of C-ELISA positive sera had evidence of recent JEV infection. Plaque reduction neutralisation tests (PRNT) using JEV, Murray Valley encephalitis (MVEV) and Kunjin (KUNV) viruses implicated JEV as the flavivirus associated with the observed antibody response in most sero-positive pigs. However, eight porcine sera with predominant neutralising antibody for KUNV (an Australasian subtype of West Nile Virus) provided evidence for the circulation of West Nile virus in Nepal. PMID- 16697906 TI - Sex, death and tragedy. AB - The population consequences of sexual conflict are relatively unexplored. In a recent paper, Le Galliard et al. now show that males of the common lizard Lacerta vivipara cause such damage to females that male-biased populations decrease in size, posing a real risk to the persistence of local lizard populations. Their study reveals surprising parallels between sexual conflict and the tragedy of the commons, where selfish competition over females destroys the very resource (i.e. the females) over which the males are fighting. PMID- 16697907 TI - Detecting anthropogenic disturbance in tropical forests. AB - Tropical forests are beleaguered by an array of threats driven by different scales of anthropogenic perturbations, which vary in the degree to which they can be detected by remote sensing. The extent of different patterns of cryptic disturbance often far exceeds the total area deforested, as shown by two recent studies on selective logging in Amazonia. Here, we discuss different forms of disturbance in Amazonian forests and question how much of the apparently intact forest in this region remains relatively undisturbed. PMID- 16697908 TI - The causes of protein evolutionary rate variation. AB - The rate of protein evolution varies more than 1000-fold and, for the past 30 years, it was thought that the rate was determined by protein function. Drummond and co-workers have now shown that a single factor underlying mRNA expression, protein abundance and synonymous codon usage is the chief causal agent of protein evolutionary rate in yeast. It will be interesting to see whether this is shown to be a universal rule for all biological systems. PMID- 16697909 TI - Disruptive selection and then what? AB - Disruptive selection occurs when extreme phenotypes have a fitness advantage over more intermediate phenotypes. The phenomenon is particularly interesting when selection keeps a population in a disruptive regime. This can lead to increased phenotypic variation while disruptive selection itself is diminished or eliminated. Here, we review processes that increase phenotypic variation in response to disruptive selection and discuss some of the possible outcomes, such as sympatric species pairs, sexual dimorphisms, phenotypic plasticity and altered community assemblages. We also identify factors influencing the likelihoods of these different outcomes. PMID- 16697910 TI - Environment and evolution through the Paleocene-Eocene thermal maximum. AB - The modern orders of mammals, Artiodactyla, Perissodactyla and Primates (APP taxa), first appear in the fossil record at the Paleocene-Eocene boundary, c. 55 million years ago. Their appearance on all three northern continents has been linked to diversification and dispersal in response to rapid environmental change at the beginning of a worldwide 100 000-200 000-year Paleocene-Eocene thermal maximum (PETM) and carbon isotope excursion. As I discuss here, global environmental events such as the PETM have had profound effects on evolution in the geological past and must be considered when modeling the history of life. The PETM is also relevant when considering the causes and consequences of global greenhouse warming. PMID- 16697911 TI - The influence of terrestrial ecosystems on climate. AB - Terrestrial ecosystems influence climate by affecting how much solar energy is absorbed by the land surface and by exchanging climatically important gases with the atmosphere. Recent model analyses show widespread qualitative agreement that terrestrial ecological processes will have a net positive feedback effect on 21st century global warming, and, therefore, cannot be ignored in climate-change projections. However, the quantitative uncertainty in the net feedback is large. The uncertainty in 21st-century carbon dioxide emissions resulting from terrestrial carbon cycle-climate feedbacks is second in magnitude only to the uncertainty in anthropogenic emissions. We estimate that this translates into an uncertainty in global warming owing to the land surface of 1.5 degrees C by 2100. We also emphasise the need to improve our understanding of terrestrial ecological processes that influence land-atmosphere interactions at relatively long timescales (decadal-century) as well as at shorter intervals (e.g. hourly). PMID- 16697912 TI - Land-plant ecology on the basis of functional traits. AB - The tissue traits and architectures of plant species are important for land-plant ecology in two ways. First, they control ecosystem processes and define habitat and resources for other taxa; thus, they are a high priority for understanding the ecosystem at a site. Second, knowledge of trait costs and benefits offers the most promising path to understanding how vegetation properties change along physical geography gradients. There exists an informal shortlist of plant traits that are thought to be most informative. Here, we summarize recent research on correlations and tradeoffs surrounding some traits that are prospects for the shortlist. By extending the list and by developing better models for how traits influence species distributions and interactions, a strong foundation of basic ecology can be established, with many practical applications. PMID- 16697913 TI - Amphioxus and tunicates as evolutionary model systems. AB - One important question in evolutionary biology concerns the origin of vertebrates from invertebrates. The current consensus is that the proximate ancestor of vertebrates was an invertebrate chordate. Today, the invertebrate chordates comprise cephalochordates (amphioxus) and tunicates (each a subphylum in the phylum Chordata, which also includes the vertebrate subphylum). It was widely accepted that, within the chordates, tunicates represent the sister group of a clade of cephalochordates plus vertebrates. However, recent studies suggest that the evolutionary positions of tunicates and cephalochordates should be reversed, the implications of which are considered here. We also review the two major groups of invertebrate chordates and compare relative advantages (and disadvantages) of each as model systems for elucidating the origin of the vertebrates. PMID- 16697914 TI - Plant-mediated effects in insect-pathogen interactions. AB - Interactions between insect herbivores and their pathogens can be modulated by host plants. Inter- and intraspecific differences in plant chemistry and structure can alter the susceptibility of insects to infection and the production and environmental persistence of pathogens. Whether plants can manipulate insect pathogens to act as "bodyguards" and increase their own fitness remains to be shown. Reduced insect performance owing to poor plant quality can enhance the susceptibility of an insect to disease while these same phytochemicals can also reduce the effectiveness of entomopathogens in killing the host. As we discuss here, plants have an important role in the evolution of insect-pathogen relationships and a tritrophic perspective should thus be incorporated into the study of insects and their pathogens. PMID- 16697915 TI - Outwit, outplay, outlive. PMID- 16697916 TI - Cell-mediated immunity in arthropods: hematopoiesis, coagulation, melanization and opsonization. AB - The functions of hemocytes in innate immune response are reviewed with emphasized on their roles in coagulation, melanization and opsonization. Also the ways in which hemocytes are produced in and released from hematopoietic tissue are discussed. PMID- 16697917 TI - Recognition of pathogens and activation of immune responses in Drosophila and horseshoe crab innate immunity. AB - In innate immunity, pattern recognition receptors discriminate between self- and infectious non-self-matter. Mammalian homologs of the Drosophila Toll protein, which are collectively referred to as Toll-like receptors (TLRs), recognize pathogen-associated molecular patterns (PAMPs), including lipopolysaccharides (LPS) and lipoproteins, whereas the Drosophila Toll protein does not act as a PAMP receptor, but rather binds to Spatzle, an endogenous peptide. In Drosophila, innate immune surveillance is mediated by members of the peptidoglycan recognition protein (PGRP) family, which recognize diverse bacteria-derived peptidoglycans and initiate appropriate immune reactions including the release of antimicrobial peptides and the activation of the prophenoloxidase cascade, the latter effecting localized wound healing, melanization, and microbial phagocytosis. In the horseshoe crab, LPS induces hemocyte exocytotic degranulation, resulting in the secretion of various defense molecules, such as coagulation factors, antimicrobial peptides, and lectins. Recent studies have demonstrated that the zymogen form of the serine protease factor C, a major granular component of hemocyte, also exists on the hemocyte surface and functions as a biosensor for LPS. The proteolytic activity of activated factor C initiates hemocyte exocytosis via a G protein mediated signal transduction pathway. Furthermore, it has become clear that an endogenous mechanism for the feedback amplification of the innate immune response exists and is dependent upon a granular component of the horseshoe crab hemocyte. PMID- 16697918 TI - Pathogen recognition and signalling in the Drosophila innate immune response. AB - Genetic analysis of the innate immune response in Drosophila has provided important insights into the mechanism of microbial sensing and the subsequent host signalling events. The two major players following immune challenge are the Toll and Immune deficiency (IMD) pathways, which are essential for fruit flies to survive infection. These pathways are homologous to the mammalian Toll-like receptor and tumour necrosis factor pathways, respectively. Moreover, microbial pattern-recognition receptors upstream of Toll and IMD, such as the peptidoglycan recognition proteins, have been isolated and studied at the structural and functional level. In the present, we will review recent data pertaining to the genetic, genomic, RNAi and infection studies that have added new complexities to the system. PMID- 16697919 TI - Proteases and protease inhibitors: a balance of activities in host-pathogen interaction. AB - The immune system is the collection of effector molecules and cells of the host that act against invading parasites and their products. Secreted proteases serve important roles in parasitic metabolism and virulence and the several families of protein protease inhibitors of the plasma and blood cells play an important role in immunity by inactivating and clearing the protease virulence factors of parasites. The protease inhibitors are of two classes, the active-site inhibitors and the alpha2-macroglobulins. Inhibitors for the first class bind and inactivate the active site of the target protease. Proteins of the second class bind proteases by a unique molecular trap mechanism and deliver the bound protease to a receptor-mediated endocytic system for degradation in secondary lysosomes. Proteins of the alpha2-macroglobulin family are present in a variety of animal phyla, including the nematodes, arthropods, mollusks, echinoderms, urochordates, and vertebrates. A shared suite of unique functional characteristics have been documented for the alpha2-macroglobulins of vertebrates, arthropods, and mollusks. The alpha2-macroglobulins of nematodes, arthropods, mollusks, and vertebrates show significant sequence identity in key functional domains. Thus, the alpha2-macroglobulins comprise an evolutionarily conserved arm of the innate immune system with similar structure and function in animal phyla separated by 0.6 billion years of evolution. PMID- 16697920 TI - Lectin complement system and pattern recognition. AB - Living organisms have strong defense mechanisms against invading microorganisms as survival strategies. One of the defense mechanisms is the complement system, composed of more than 30 serum and cell surface components. This system collaborates in recognition and elimination of pathogens as a part of both the innate and acquired immune systems. The two collagenous lectins, mannose-binding lectin (MBL) and ficolins, are pattern recognition proteins acting in innate immunity and, upon recognition of the pathogens, they trigger the activation of the lectin complement pathway through attached serine proteases (MASPs). A similar lectin-based complement system, consisting of the lectin-protease complex and C3, is present in ascidians, our closest invertebrate relatives and in lamprey, the most primitive vertebrate. Furthermore, a lamprey N acetylglucosamine (GlcNAc)-binding lectin was identified as the orthlogue of mammalian C1q, and lamprey MASP is suggested as the prototype of MASP-2/C1r/C1s, indicating that the classical complement pathway arose as a part of the innate immune system. Thus, the complement system is one of the most highly organized innate immune systems in invertebrates and jawless vertebrates, and this system has survived in vertebrates with its core components little changed for 600-700 million years. PMID- 16697921 TI - Iron-withholding strategy in innate immunity. AB - The knowledge of how organisms fight infections has largely been built upon the ability of host innate immune molecules to recognize microbial determinants. Although of overwhelming importance, pathogen recognition is but only one of the facets of innate immunity. A primitive yet effective antimicrobial mechanism which operates by depriving microbial organisms of their nutrients has been brought into the forefront of innate immunity once again. Such a tactic is commonly referred to as the iron-withholding strategy of innate immunity. In this review, we introduce various vertebrate iron-binding proteins and their invertebrate homologues, so as to impress upon readers an obscured arm of innate immune defense. An excellent comprehension of the mechanics of innate immunity paves the way for the possibility that novel antimicrobial therapeutics may emerge one day to overcome the prevalent antibiotic resistance in bacteria. PMID- 16697922 TI - Host defence peptides from invertebrates--emerging antimicrobial strategies. AB - Cationic antimicrobial (host defence) peptides are found as potent components of the innate immune system of all invertebrates in which they have been investigated. They vary substantially in their amino acid sequences, secondary structures, inducibility, potency and antimicrobial activity spectra. This enormous diversity is providing templates for the design and development of both antibiotic peptides and peptides that selectively modulate innate immunity to increase protection against infections and sepsis. PMID- 16697924 TI - Percutaneous therapies for valvular heart disease. AB - Balloon valvuloplasty has been in clinical use for over 20 years, but the prospect of repairing and replacing cardiac valves via catheter-based techniques represents a truly recent development. This review introduces evolving technologies and their relevance to cardiovascular pathologists. PMID- 16697925 TI - Changes in cardiovascular function on treatment of inhibitors of apoptotic signal transduction pathways in left ventricular remodeling after myocardial infarction. AB - INTRODUCTION: It has been suggested that apoptosis in cardiac remodeling after myocardial infarction (MI) occurs in cardiomyocytes and is critically involved in the process of postinfarct cardiac remodeling. We investigated the pathophysiological link between myocardial apoptosis and cardiovascular function by modulating apoptotic signal transduction pathways. METHODS: Either a caspase-3 inhibitor (CasI) or a calpain inhibitor (CalI) was administered immediately after MI in a rat model of MI. Blood pressure (BP), heart rate (HR), and blood flow velocity (BFV) were measured, and pressure-rate product (PRP) was calculated to estimate the changes in cardiovascular function (n=6 for each group). RESULTS: BFV showed no remarkable changes in any of the groups. Both systolic blood pressure (SBP) and HR changed significantly (P<.01) in the MI+CasI and MI+CalI groups at 1 day after MI and returned to control levels thereafter. In contrast, SBP and HR remained significantly (P<.01) altered in the MI group. PRP in the MI groups was significantly decreased (P<.05 in the MI and MI+CasI groups; P<.01 in the MI+CalI group) at 1 day after MI and returned to control levels at 4 days. CONCLUSION: This study suggests that inhibition of apoptosis during left ventricular remodeling ameliorates cardiovascular function in remodeled hearts. PMID- 16697927 TI - Serum biomarkers for heart failure. AB - The incidence of heart failure has been steadily increasing over the past several decades. High readmission rates in patients with acute decompensated heart failure led to the search for biomarkers that could predict future clinical course and would, in an ideal case, enable monitoring of patients with heart failure and guidance of their therapy. From among several promising markers, the B-type natriuretic peptide and the biologically inactive N-terminal portion of its pro-hormone, NT-proBNP, have become the most frequently used analytes. Other known markers, such as atrial natriuretic peptide and endothelin-1, are currently used for research purposes. The development of additional biomarkers will be an important step from improving diagnosis and treatment of patients with chronic and acute decompensated heart failure. PMID- 16697926 TI - Relevance of inflammatory cell infiltrates for complete atrioventricular block in experimental murine myocarditis. AB - BACKGROUND: There are few systemic pathologic studies on myocarditis. This study aimed to clarify the pathologic characteristics of murine myocarditis. METHODS: We recorded serial electrocardiograms in experimental viral myocarditis in mice and then examined their cardiac pathology. After taking baseline electrocardiograms, we inoculated the mice intraperitoneally with the encephalomyocarditis virus. Electrocardiograms were serially recorded until 220 days after the virus inoculation. RESULTS: Serial electrocardiograms revealed ectopic beats, low voltage of the QRS complex, and the appearance of complete atrioventricular (AV) block. Corresponding myocardial lesions were found in the hearts of mice with these ectopic beats. Mononuclear cell infiltrations into the His bundle were most frequently found in mice with complete AV block. CONCLUSIONS: Inflammatory change with cellular infiltrations was the most common pathologic finding in mice with complete AV block. In clinical settings, anti inflammatory therapy might be recommended for patients with myocarditis complicated with conduction disturbances. PMID- 16697928 TI - A giant coronary artery aneurysm in the right coronary artery. AB - Aneurysms of the coronary arteries are uncommon occurrences that usually develop secondary to atherosclerosis and are often asymptomatic. They are usually diagnosed incidentally during investigation for ischemic heart disease or at autopsy for sudden death. We present a case of a "giant" right coronary artery aneurysm (CAA) discovered incidentally at surgery. Pathological examination confirmed that this was a true aneurysm showing marked thinning of the media and fibrocalcific plaques with small, multifocal areas of lymphocytic infiltrates. PMID- 16697929 TI - Rupture of the thyrocervical trunk branch from the subclavian artery in a patient with neurofibromatosis: a case report. AB - BACKGROUND: Vascular involvement in neurofibromatosis type 1 (NF1) is well recognized; however, rupture of extracranial arteries rarely occurs. We present a case of NF1 with rupture of the thyrocervical trunk, which branched from the right subclavian artery. A 76-year-old woman who has numerous cafe-au-lait spots and soft tumors of the skin manifested a sudden swelling of her neck accompanied with increasing pain. Radiological examinations revealed bleeding from the artery. METHODS: Histological and immunohistochemical examinations were carried out using tissues that contained the affected vessel. RESULTS: Proliferation of spindle cells positive for S-100 protein was seen in the adventitia of the ruptured vessel. Intimal thickening by proliferation of fibromuscular cells was also evident with irregularity of the media. CONCLUSIONS: These findings suggest that the artery was disrupted by NF in the vascular wall. It is considered that NF in the arterial wall causes dysplasia of the smooth muscle layer in the intima and media and leads to fragility of the vessel. Twelve cases, including the present case, with rupture of extracranial arteries in NF1 have been reported in the past 10 years; two thirds of these occurred in extravisceral sites in which there is a good deal of physical movement. This suggests that a physiological factor is one of the triggers for arterial rupture, which occurs under a background of vascular fragility in NF1. PMID- 16697930 TI - Fatal cor pulmonale caused by pulmonary tumor microembolism in a patient with occult gastric cancer. AB - BACKGROUND: A fatal pulmonary tumor microembolism is rarely caused by an occult gastric cancer. METHODS AND RESULTS: We report the case of a 40-year-old woman who died after 3 days of progressive dyspnea, cough, and pulmonary hypertension. Postmortem examination demonstrated the presence of an occult diffuse-type gastric carcinoma, which had caused emboli in about 80% of small pulmonary arteries and arterioles. Despite an interatrial defect in the fossa ovalis, no parenchymal metastases were documented. CONCLUSION: Pulmonary tumor microembolism may be suspected in patients complaining of unexplained progressive dyspnea and who develop acute or subacute cor pulmonale. PMID- 16697931 TI - Case report of a Quilty lesion within a coronary artery. AB - The Quilty lesion is the moniker of a lymphoid proliferation that resides predominantly in the endocardium and underlying myocardium within transplanted hearts. It has been described in the endocardium, myocardium, and epicardium. Although its pathogenesis is not understood, it is generally considered that the Quilty lesion is not a manifestation of acute rejection. However, its clinical significance and relation to acute rejection, such as its pathogenesis, are still not completely understood. Immunohistochemical studies distinguish between the cellular infiltrates of Quilty lesions and those associated with acute rejection. We describe, what we believe to be, the first case of a Quilty lesion in an epicardial coronary artery from an 8-year-old girl with congenital heart disease who required retransplantation 25 months after her cardiac transplantation for severe graft coronary artery disease. The lesion consisted of an intramural nodular aggregate of numerous T and B lymphocytes, with fewer macrophages, characteristic of a Quilty lesion. PMID- 16697932 TI - Calcific bicuspid aortic valve disease in a patient with Cornelia de Lange syndrome: linking altered Notch signaling to aortic valve disease. AB - We describe a patient with Cornelia de Lange syndrome (CdLS) and a severely calcified bicuspid aortic valve. Cornelia de Lange syndrome is characterized by altered Notch signaling, and recent studies have provided a link between Notch signaling and heart valve development and calcific bicuspid aortic valve disease. In this case report, we propose that altered Notch signaling in CdLS may be causally linked to the calcific bicuspid aortic valve disease in these patients. Patients with CdLS should undergo routine echocardiographic examination for possible congenital cardiac defects including bicuspid aortic valve. PMID- 16697933 TI - Black pigmentation of atherosclerotic plaques associated with chronic minocycline therapy. AB - Minocycline is a broad-spectrum antimicrobial agent used chronically in many clinical settings. Pigmentation is a well-recognized side effect of minocycline therapy. It has been reported in numerous sites, especially the skin, bones, and thyroid gland. We report an unusual case of pigmentation of atherosclerotic plaques in the aorta and arteries of the leg in a patient on chronic minocycline therapy. PMID- 16697934 TI - Leiomyosarcoma of the inferior vena cava. AB - Leiomyosarcoma of the inferior vena cava is a rare neoplasm; however, it is the most frequent tumor of vascular origin. It is classified according to site of origin that reflects its clinical presentation and prognosis. They are generally recognized to be of smooth muscle cell origin. Surgical resection is the treatment of choice. The role of adjuvant therapy for leiomyosarcoma is not established. PMID- 16697935 TI - Myocardial infarction with spontaneous reperfusion in a young woman with hypertrophic cardiomyopathy. PMID- 16697936 TI - A rationale for a stone on the heart--subepicardial lipoma. PMID- 16697937 TI - Design and conduct of clinical trials in patients with osteoarthritis of the hand: recommendations from a task force of the Osteoarthritis Research Society International. PMID- 16697939 TI - Chronic exposure to intra-amniotic lipopolysaccharide affects the ovine fetal brain. AB - OBJECTIVE: Fetal brain injury is associated with chorioamnionitis, which is often present without signs of overt infection or fetal compromise. We aimed to determine if prolonged exposure to intrauterine inflammation caused by intra amniotic infusion of lipopolysaccharide (LPS) would affect the fetal brain. METHODS: At 80 days of pregnancy ewes bearing singletons had osmotic pumps implanted intra-amniotically to infuse Escherichia coli LPS (055:B5; n = 8) or saline (n = 7) for 28 days. At delivery (110 days), umbilical arterial blood and chorioamnion were assessed for inflammation; cytokine concentrations (interleukin [IL]-6 and IL-8) in amniotic fluid and fetal and maternal plasma were measured. The fetal cerebral hemispheres were examined for gross anatomical changes and the number of activated microglia/macrophages, astrocytes, and oligodendrocytes estimated after immunohistochemical staining. RESULTS: Intra-amniotic administration of LPS caused chorioamnionitis, fetal leucocytosis, and a moderate to extensive infiltration of activated microglia/macrophages in the subcortical white matter in six of eight fetuses; the remaining two fetuses were less affected. Within these focal regions of damage there was an attenuation of astrocytic processes, axonal injury, and a reduction in the number of 2',3' cyclic nucleotide 3'-phosphodiesterase (CNPase) immunoreactive oligodendrocytes in areas of extensive focal damage. In control fetuses there was mild (3/7) or no infiltration of activated microglia/macrophages in the subcortical white matter. Overall the infiltration of activated microglia/macrophages in the white matter was significantly greater in LPS-exposed fetuses compared to controls. In regions devoid of injury, the number of oligodendrocytes and astrocytes was not different between groups, nor was there a difference in the volume of cerebral white matter or density of blood vessels within the white matter. Amniotic fluid IL-6 and IL 8, and maternal plasma IL-8 concentrations were significantly increased by LPS infusion. CONCLUSIONS: An increase in inflammatory cells and axonal disruption in the subcortical white matter of the fetal brain can accompany chorioamnionitis induced by intra-amniotic administration of LPS, but cystic lesions do not occur. Thus, the effect on the fetal brain is milder than that reported from animal models of acute fetal/intrauterine infection. PMID- 16697940 TI - Effects of multiple courses of antenatal corticosteroids on blood-brain barrier permeability in the ovine fetus. AB - OBJECTIVE: To test the hypothesis that multiple courses of antenatal corticosteroids accentuate the decreases in blood-brain barrier permeability observed after a single course of corticosteroids in preterm ovine fetuses. METHODS: Chronically instrumented 106-day gestation ovine fetuses were studied after single and multiple courses of dexamethasone or placebo were given to ewes beginning at 104 to 106 or 76 to 78 days of gestation, respectively. In the single-course groups, the ewes received dexamethasone (6 mg, n = 6) or placebo (n = 6) as four intramuscular injections every 12 hours over 48 hours. In the multiple course groups, the ewes received the same treatment (dexamethasone, n = 9, or placebo, n = 8), once per week for 5 weeks starting at 76 to 78 days of gestation. Blood-brain barrier permeability was quantified with the blood-to brain transfer constant (K(i)) for alpha-aminoisobutyric acid (AIB) in the brain regions of the fetuses 12 hours after the last injection of dexamethasone was given to the ewes at 106 to 107 days of gestation. RESULTS: Both single (analysis of variance [ANOVA]; main effects for dexamethasone treatment, F = 5.92, P <.04) and multiple (ANOVA; main effects for dexamethasone treatment, F = 4.74, P <.04) courses of antenatal corticosteroids were associated with decreases in blood brain barrier permeability in the brain regions of the ovine fetus. However, the multiple courses did not accentuate (ANOVA; main effects for single versus multiple courses, F = 1.06, P = .32) the decreases in permeability observed after a single course. CONCLUSION: Contrary to our hypothesis, antenatal treatment with a 5-week course of corticosteroids did not accentuate the reductions in blood brain barrier permeability that we observed after a single course of corticosteroids in the fetus. PMID- 16697941 TI - Gene expression patterns in the developing murine placenta. AB - OBJECTIVE: Successful placental development is crucial for optimal growth, maturation, and survival of the embryo/fetus. To examine genetic aspects of placental development, we investigated gene expression patterns in the murine placenta at embryonic day 10.5 (E10.5), E12.5, E15.5, and E17.5. METHODS: By use of the Affymetrix MU74A array (Affymetrix, Santa Clara, CA), we measured expression levels for 12,473 probe sets. Using pairwise analysis we selected 622 probe sets, corresponding to 599 genes, that were up- or down-regulated by more than fourfold between time points E10.5 and E12.5, E12.5 and E15.5, E15.5 and E17.5. We analyzed and functionally annotated those genes regulated during development. RESULTS: In comparing E10.5 to E12.5 we found that angiogenesis and fatty acid metabolism and transport related genes were up-regulated at E10.5, while genes involved in hormonal control and ribosomal proteins were up-regulated at E12.5. When comparing E12.5 to E15.5 we noted that genes involved in the cell cycle and RNA metabolism were strongly up-regulated at E12.5, while genes involved in cellular transport were up-regulated at E15.5. Finally, when comparing E15.5 to E17.5, we found genes related to cell cycle control, genes expressed in the nucleus and involved in RNA metabolism were up-regulated at E17.5. CONCLUSION: Microarray analysis has allowed us to describe gene expression patterns and profiles in the developing mouse placenta. Further analysis has demonstrated that several functional classes are up- and down-regulated at specific time points in placental development. These changes may have significant implications for placental development in the human. PMID- 16697942 TI - Macrophage migration inhibitory factor-nitric oxide interaction in human fetal membranes at term pregnancy. AB - OBJECTIVES: Macrophage migration inhibitory factor (MIF), a multifunctional proinflammatory cytokine, has been recently involved in many aspects of reproduction including pregnancy. However, no evidence is available on the role of MIF in gestational tissues nor on factors regulating MIF production. This study, conducted on explants of human fetal membranes at term gestation, has been undertaken to investigate whether: (1) MIF is produced by fetal membranes; (2) nitric oxide (NO) can regulate local MIF production; and (3) MIF, in turn, can influence NO release in these tissues. METHODS: Tissues were obtained from 56 healthy women who underwent elective cesarean delivery. Fetal membranes have been incubated with either sodium nitroprusside (NP), a NO donor, or recombinant MIF (r-MIF), or a specific anti-MIF antibody (MIF-Ab). Reverse transcriptase polymerase chain reaction (RT-PCR), Western blot, enzyme-linked immunosorbent assay (ELISA), and colorimetric assay have been used to detect MIF mRNA and protein, inducible nitric oxide synthase (iNOS), and NO metabolites. RESULTS: Fetal membranes basally express MIF mRNA and protein and release MIF. Exposing tissues to NP results in an increase of MIF mRNA expression and protein release. Conversely, treatment of tissues with MIF is followed by a reduction in iNOS mRNA and protein expression as well as in NO release. These effects are reversed by adding MIF-Ab. CONCLUSIONS: MIF is generated and released by human fetal membranes at term. MIF mRNA and protein expression and release are modulated by NO. MIF, in turn, can reduce iNOS expression and NO release by these tissues. NO could be a regulator of MIF production in pregnancy and labor. PMID- 16697943 TI - Differential secretion of matrix metalloproteinase-2 and -9 after selective infection with group B streptococci in human fetal membranes. AB - OBJECTIVE: This study evaluated the secretions of zymogen and active forms of matrix metalloproteinase (MMP)-9 and MMP-2 and their specific inhibitors, TIMP-1 and TIMP-2 by fetal membranes stimulated with group B Streptoccocci (GBS). METHODS: We used an in vitro experimental model that allowed us to estimate the individual contribution of the amnion (AM) and the choriodecidua (CHD) to the microbial insult. Membranes were obtained after delivery by elective cesarean delivery from women at 37 to 40 weeks of gestation without evidence of either active labor or intrauterine infection. Membranes were mounted in Transwell devices (Costar, New York, NY), physically separating the upper and lower chambers; 1 x 10(6) CFU of GBS was added to either AM or CHD and the secretions and gelatinolytic activity of MMP-2 and MMP-9 were measured in both compartments by enzyme-linked immunosorbent assay (ELISA) and zymography. TIMPs secretion was measured by ELISA. Both MMPs were immunolocalized in tissue sections. RESULTS: The simultaneous stimulation at both sides was followed by increases of proMMP-9 (85.0 +/- 18.63 pg/mL) and proMMP-2 (4.10 +/- 1.90 ng/mL) in the CHD (P <.05). When only one side of the membrane was stimulated, the secretion level of proMMP 2 increased 2.3-fold and that of proMMP-9 2.5-fold in the CHD. The active forms of both enzymes did not change with any modality of stimulation. The secretion level of both TIMPs remained without significant changes. CHD and AM were positive for immunoreactive MMP-2 and MMP-9. CONCLUSION: We propose that infection of fetal membranes with GBS is followed by active secretion of MMP and the CHD is the principal source of these mediators of extracellular matrix degradation. PMID- 16697944 TI - Association of fetal choroid plexus cysts with trisomy 18 in a population previously screened by nuchal translucency thickness measurement. AB - OBJECTIVE: The study's aim was to determine any association between choroid plexus cysts (CPCs) and trisomy 18 in a population of fetuses previously screened by nuchal translucency (NT) thickness measurement. METHODS: During the study period (May 1999 to December 2004), 7,795 fetuses had an NT scan and second trimester fetal anatomical scan at our institution. The prevalence of trisomy 18 was determined among four types of pregnancies: 1) those with isolated CPCs, 2) those with CPCs and enlarged NT, 3) those with CPCs and other ultrasound markers, and 4) those with CPCs, enlarged NT, and other ultrasound markers. The fetal outcome according to NT and presence of CPCs was calculated. Incidence rates of enlarged NT and CPCs in fetuses with trisomy 18 and fetuses with normal chromosomes were also evaluated. RESULTS: For the entire population, ten trisomy 18 cases were diagnosed prenatally (prevalence, 0.13%). Among fetuses with enlarged NT, the likelihood ratio of trisomy 18 was significantly increased in fetuses with CPCs compared with fetuses without such cysts (333.6 versus 15.2, P = .002). However, among fetuses with normal NT, no significant difference was demonstrated for likelihood ratio of trisomy 18 between fetuses with and without CPCs. CONCLUSION: In pregnancies complicated by isolated CPCs, fetal karyotyping is not indicated when no additional anomaly is detected on ultrasonographic examination and first-trimester NT results are normal. PMID- 16697946 TI - The depolarized expression of the alpha-6 integrin subunit in the endometria of women with endometriosis. AB - OBJECTIVE: The current study sought to compare the endometrial localization of the integrin subunit alpha-6 in women with endometriosis and women without the disease. Alpha-6 integrins have an important function, not only in the attachment of cells to the extracellular matrix and laminin, but they also serve as inductors of cell migration and invasion, depending on their pattern of expression in the cell membrane. METHODS: The endometriosis group consisted of 32 women with a confirmed diagnosis of endometriosis by laparoscopy or laparotomy. The control group consisted of 20 women not having endometriosis or any other gynecologic disease at laparoscopy. Endometria were obtained by biopsy. Immunohistochemical techniques were used to assess alpha-6 localization. In each section, the percentage of positive cells and the localization of expression were evaluated. RESULTS: All glandular cells expressed alpha-6 in all of the samples but presented two different patterns, either only in the basal side of the cells (polarized) or also in other sides of the cells (depolarized). The percentage of samples showing depolarized expression was significantly higher in the endometriosis group (66.6% vs 15.8%, chi2 =12.09, P = .001). CONCLUSIONS: The endometria of women with endometriosis more frequently show a depolarized expression of integrin subunit alpha-6, a characteristic usually found in highly proliferating cells with migrating and invasive abilities. PMID- 16697945 TI - Analysis of free hemoglobin level and hemoglobin peptides from human puerperal uterine secretions. AB - OBJECTIVE: Hemocidins are a novel class of antibacterial peptides generated proteolytically from hemoglobin. These peptides play a particularly important role in maintaining vaginal homeostasis during menstrual bleeding. To investigate the hemoglobin fragmentation process during the last stages of pregnancy, we examined uterine secretion (lochia) samples from a group of 22 healthy women who underwent cesarean delivery at term. METHODS: Patients were divided into three groups: (1) the elective cesarean deliveries without symptoms of spontaneous labor, (2) the nonelective cesarean deliveries with spontaneous beginning of labor, and (3) the nonelective cesarean deliveries during advanced labor. The samples were subjected to chromatographic estimation of free hemoglobin and peptides. In three representative patients the identification of all lochial peptides was performed. RESULTS: All samples contained a significant amount of free hemoglobin and its level increased with labor progression. The presence of peptide fractions was also detected in most lochia samples. They were confirmed to be human hemoglobin fragments, almost identical to the recently described bactericidal hemocidins from menstrual discharge. The level of peptides also increased during labor. The subgroup with advanced labor demonstrated the highest amount of hemocidins. CONCLUSIONS: The presented results prove that proteolysis of free hemoglobin in the female upper reproductive tract begins together with the clinical symptoms of normal labor. We speculate that cesarean delivery affects molecular mechanisms involved in antibacterial hemocidins generation and, in effect, might be responsible for the increased risk of gynecologic infections in cesarean deliveries. PMID- 16697947 TI - Preoperative treatment of uterine leiomyomas: clinical findings and expression of transforming growth factor-beta3 and connective tissue growth factor. AB - OBJECTIVE: To evaluate the clinical features and the expression of transforming growth factor-beta3 (TGF-beta3) and connective tissue growth factor (CTGF) in myometrium and uterine leiomyomas after preoperative treatment with gonadotropin releasing hormone-analogs (GnRH-a) and tibolone. METHODS: Twenty-three patients received 3.75 mg leuprolide acetate depot for 4 months. Twenty-two patients received the same therapy plus 2.5 mg tibolone daily. Patients underwent uterine surgery after therapy. Twenty-two untreated patients underwent surgery directly. Hematologic tests, bone mineral density (BMD) measurement, and ultrasonographic evaluation of uterine volume were performed before and after treatment. Menorrhagia and pelvic pain were evaluated with a visual analog scale. Hot flushes were recorded in daily diaries. Immunohistochemical expression of TGF beta3 and CTGF in myometrium and myoma samples was evaluated semiquantitatively. RESULTS: After therapy, hemoglobin and iron levels similarly increased in both groups. BMD significantly decreased only in the GnRH-a group. Uterine volume similarly decreased in both groups. No patient had menorrhagia or pelvic pain at the end of therapy. The number of hot flushes increased after the first month in the GnRH-a group; in the GnRH-a plus tibolone group, it remained constant and was lower. In untreated cases, TGF-beta3 and CTGF smooth muscle cell immunoexpression was lower in myometrium than in leiomyomas. After medical treatment, growth factor immunoexpression remained unchanged in myometrial samples and was reduced in leiomyomas. Endothelial cells showed strong immunopositivity, both in untreated and in treated cases. CONCLUSION: This study focuses on the effects of GnRH-a and tibolone on TGF-beta3 and CTGF expression in myometrium and myomas and supports the hypothesis of a pathogenetic role of these growth factors in uterine fibromatosis. PMID- 16697948 TI - The granulin-epithelin precursor is a steroid-regulated growth factor in endometrial cancer. AB - OBJECTIVES: The majority of endometrial cancers arise as a result of estrogen stimulation, the molecular targets of which remain incompletely defined. We hypothesize that the granulin-epithelin precursor (GEP) may be one such target. In this study, we examined the frequency of GEP and estrogen receptor (ER) co expression in human endometrial cancers. Once we established the co-expression of GEP with the estrogen receptor we examined the potential estrogen regulation of GEP expression, as well as the functional significance of GEP expression in vitro. METHODS: Double immunofluorescence and confocal microscopy were used to compare GEP and ER expression among 41 endometrial cancers. The effects of estradiol and tamoxifen treatment on GEP expression in two endometrial cancer cell lines, KLE and HEC-1-A, were assessed through reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot analysis. The antiproliferative effect of GEP silencing by short hairpin (sh)RNA, was evaluated in HEC-1-A cells using an MTT assay. RESULTS: GEP co-expression with ER was observed in 63% of the cancers examined. A two- to fivefold increase in GEP expression with estradiol and/or tamoxifen treatment was observed in KLE cells. Silencing of GEP in HEC-1-A cells using shRNA resulted in a decrease in proliferation among transfected cells. CONCLUSIONS: Co-expression of GEP and ER in endometrial cancer cells, and the regulation of GEP by estrogen, suggests a role for GEP in steroid-mediated endometrial cancer cell growth. Further characterization of GEP as a steroid-mediated growth factor in these cells may broaden our understanding of endometrial cancer biology and also provide guidance in the development of novel therapeutic targets. PMID- 16697949 TI - Potassium channels in the vasodilating action of levosimendan on the human umbilical artery. AB - OBJECTIVE: Levosimendan is a calcium-sensitizing agent and inodilator working via potassium channels, which is under current investigation in the treatment of heart failure. We investigated the type of potassium channels that play a role on the dilatating effect of levosimendan on the contractile tones of the isolated human umbilical artery (HUA). METHODS: The response in the HUA was recorded isometrically by a force displacement transducer in isolated organ baths. Levosimendan was added to organ baths after precontraction with serotonin (5-HT, 1 microM). Levosimendan-induced relaxations were tested in the presence of the large conductance Ca2+-activated K+ channel inhibitor tetraethylammonium (TEA, 1 mM), the adenosine triphosphate (ATP)-sensitive K+ channel inhibitor glibenclamide (GLI, 10 microM), and the voltage-sensitive K+ channel inhibitor 4 aminopyridine (4-AP, 1 mM). All experiments were performed in solutions containing the cyclooxygenase inhibitor indomethacin (10 microM) and the nitric oxide synthase inhibitor L-NAME (100 microM). RESULTS: Levosimendan (10 nM to 3 microM) produced potent relaxation in the HUA. Vehicle had no significant relaxant effect. The relaxation to levosimendan was not affected by the K+ channel inhibitor, GLI. However, 4-AP (1 mM) and TEA (1 mM) inhibited levosimendan-induced relaxation significantly (P <.05). CONCLUSION: These results show that levosimendan effectively and directly decreases the tone of the HUA. The mechanism of this levosimendan-induced relaxation in the HUA appears in part to be due to voltage-gated and large conductance Ca2+-activated K+ channel opening action. PMID- 16697950 TI - The systematic review of systematic reviews has arrived! PMID- 16697951 TI - Kinase inhibitors: vice becomes virtue. AB - In this issue of Cancer Cell, Fan and coworkers describe a novel inhibitor of PI3 kinase (PI3K) that potently interferes with the growth of glioma cells. They show that the efficacy of this inhibitor results from dual, synergistic activity against the p110alpha subunit of PI3K and against TOR. Although p110alpha and TOR belong to the same signaling pathway, they both must be inactivated because of the need to silence the regulatory feedback loop that remains unaffected by monospecific inhibitors. The new PI3K inhibitor achieves the effects of combination therapy as a single agent by fortuitously hitting two critical targets. PMID- 16697952 TI - At the gates of death. AB - Apoptosis that proceeds via the mitochondrial pathway involves mitochondrial outer membrane permeabilization (MOMP), responsible for the release of cytochrome c and other proteins of the mitochondrial intermembrane space. This essential step is controlled and mediated by proteins of the Bcl-2 family. The proapoptotic proteins Bax and Bak are required for MOMP, while the antiapoptotic Bcl-2 proteins, including Bcl-2, Bcl-xL, Mcl-1, and others, prevent MOMP. Different proapoptotic BH3-only proteins act to interfere with the function of the antiapoptotic Bcl-2 members and/or activate Bax and Bak. Here, we discuss an emerging view, proposed by Certo et al. in this issue of Cancer Cell, on how these interactions result in MOMP and apoptosis. PMID- 16697953 TI - Ewing's sarcoma: general insights from a rare model. AB - Ewing's sarcoma is characterized by the presence of fusion oncoproteins involving EWSR1 and an ETS gene, most commonly FLI1. In this issue of Cancer Cell, Smith et al. have combined RNA interference with expression profiling to study the pattern of gene expression downstream of the most common of these fusions, EWS/FLI. Using this strategy, Smith et al. have identified a homeobox gene, NKX2.2, which is both highly expressed in Ewing's sarcoma and essential for the transforming activity of EWS/FLI. PMID- 16697954 TI - The challenges of gene expression microarrays for the study of human cancer. AB - Large-scale genomic studies promise to advance our understanding of the biology of human cancers and to improve their diagnosis, prognostication, and treatment. The analysis and interpretation of genomics studies have faced challenges. The retrospective and observational design of many studies has rendered them susceptible to confounding and bias. Technological variations and advances have impacted on reproducibility. Statistical hurdles in relating a large number of variables to a small number of observations have added further constraints. This review considers the promise and challenge associated with the large-scale clinically oriented genomic analysis of human cancer and attempts to emphasize potential solutions. PMID- 16697955 TI - A dual PI3 kinase/mTOR inhibitor reveals emergent efficacy in glioma. AB - The PI3 kinase family of lipid kinases promotes cell growth and survival by generating the second messenger phosphatidylinositol-3,4,5-trisphosphate. To define targets critical for cancers driven by activation of PI3 kinase, we screened a panel of potent and structurally diverse drug-like molecules that target this enzyme family. Surprisingly, a single agent (PI-103) effected proliferative arrest in glioma cells, despite the ability of many compounds to block PI3 kinase signaling through its downstream effector, Akt. The unique cellular activity of PI-103 was traced directly to its ability to inhibit both PI3 kinase alpha and mTOR. PI-103 showed significant activity in xenografted tumors with no observable toxicity. These data demonstrate an emergent efficacy due to combinatorial inhibition of mTOR and PI3 kinase alpha in malignant glioma. PMID- 16697956 TI - Mitochondria primed by death signals determine cellular addiction to antiapoptotic BCL-2 family members. AB - We show that the antiapoptotic proteins BCL-2, BCL-XL, MCL-1, BFL-1, and BCL-w each bear a unique pattern of interaction with a panel of peptides derived from BH3 domains of BH3-only proteins. Cellular dependence on an antiapoptotic protein for survival can be decoded based on the pattern of mitochondrial sensitivity to this peptide panel, a strategy that we call BH3 profiling. Dependence on antiapoptotic proteins correlates with sequestration of activator BH3-only proteins like BID or BIM by antiapoptotic proteins. Sensitivity to the cell permeable BCL-2 antagonist ABT-737 is also related to priming of BCL-2 by activator BH3-only molecules. Our data allow us to distinguish a cellular state we call "primed for death," which can be determined by BH3 profiling and which correlates with dependence on antiapoptotic family members for survival. PMID- 16697957 TI - NKX3.1 stabilizes p53, inhibits AKT activation, and blocks prostate cancer initiation caused by PTEN loss. AB - We demonstrate that PTEN loss causes reduced NKX3.1 expression in both murine and human prostate cancers. Restoration of Nkx3.1 expression in vivo in Pten null epithelium leads to decreased cell proliferation, increased cell death, and prevention of tumor initiation. Whereas androgen receptor (AR) positively regulates NKX3.1 expression, NKX3.1 negatively modulates AR transcription and consequently the AR-associated signaling events. Consistent with its tumor suppressor functions, NKX3.1 engages cell cycle and cell death machinery via association with HDAC1, leading to increased p53 acetylation and half-life through MDM2-dependent mechanisms. Importantly, overexpression of Nkx3.1 has little effect on Pten wild-type epithelium, suggesting that PTEN plays a predominant role in PTEN-NKX3.1 interplay. Manipulating NKX3.1 expression may serve as a therapeutic strategy for treating PTEN-deficient prostate cancers. PMID- 16697958 TI - The PTEN and INK4A/ARF tumor suppressors maintain myelolymphoid homeostasis and cooperate to constrain histiocytic sarcoma development in humans. AB - Histiocytic sarcoma (HS) is a rare malignant proliferation of histiocytes of uncertain molecular pathogenesis. Here, genetic analysis of coincident loss of Pten and Ink4a/Arf tumor suppressors in the mouse revealed a neoplastic phenotype dominated by a premalignant expansion of biphenotypic myelolymphoid cells followed by the development of HS. Pten protein loss occurred only in the histiocytic portion of tumors, suggesting a stepwise genetic inactivation in the generation of HS. Similarly, human HS showed genetic or epigenetic inactivation of PTEN, p16(INK4A), and p14(ARF), supporting the relevance of this genetically engineered mouse model of HS. These genetic and translational observations establish a cooperative role of Pten and Ink4a/Arf in the development of HS and provide mechanistic insights into the pathogenesis of human HS. PMID- 16697959 TI - Tumor stem cells derived from glioblastomas cultured in bFGF and EGF more closely mirror the phenotype and genotype of primary tumors than do serum-cultured cell lines. AB - The concept of tumor stem cells (TSCs) provides a new paradigm for understanding tumor biology, although it remains unclear whether TSCs will prove to be a more robust model than traditional cancer cell lines. We demonstrate marked phenotypic and genotypic differences between primary human tumor-derived TSCs and their matched glioma cell lines. Unlike the matched, traditionally grown tumor cell lines, TSCs derived directly from primary glioblastomas harbor extensive similarities to normal neural stem cells and recapitulate the genotype, gene expression patterns, and in vivo biology of human glioblastomas. These findings suggest that TSCs may be a more reliable model than many commonly utilized cancer cell lines for understanding the biology of primary human tumors. PMID- 16697960 TI - Expression profiling of EWS/FLI identifies NKX2.2 as a critical target gene in Ewing's sarcoma. AB - Our understanding of Ewing's sarcoma development mediated by the EWS/FLI fusion protein has been limited by a lack of knowledge regarding the tumor cell of origin. To circumvent this, we analyzed the function of EWS/FLI in Ewing's sarcoma itself. By combining retroviral-mediated RNA interference with reexpression studies, we show that ongoing EWS/FLI expression is required for the tumorigenic phenotype of Ewing's sarcoma. We used this system to define the full complement of EWS/FLI-regulated genes in Ewing's sarcoma. Functional analysis revealed that NKX2.2 is an EWS/FLI-regulated gene that is necessary for oncogenic transformation in this tumor. Thus, we developed a highly validated transcriptional profile for the EWS/FLI fusion protein and identified a critical target gene in Ewing's sarcoma development. PMID- 16697962 TI - Autoimmunity in the era of genomics and proteomics. AB - Autoimmune diseases arise from an aberrant recognition of self-epitopes by the immune system, resulting in inflammation and tissue damage. The wide range of affected organ systems and the overlap between different diseases pose several clinical challenges. Apart from establishing an accurate diagnosis, some of the problems frequently encountered are assessment of risk factors, prediction of prognosis and administration of an optimal treatment regimen. In recent years, the introduction of novel laboratory techniques have raised hopes that new answers will be provided to these challenges. In this work, we have summarized some of the prominent contributions of genetic works, microarray analyses and autoantigen microarrays to the understanding of autoimmunity and management of patients. PMID- 16697963 TI - Protein arrays and pattern recognition: new tools to assist in the identification and management of autoimmune disease. AB - The occurrence of antibody patterns in connective tissue diseases has been recognized for thirty years, but the generation of multiple antibody results relied on time-consuming immunodiffusion or electrophoretic techniques. Today it is possible to study the antibody repertoire using rapid multi-analyte technologies, generally referred to as protein arrays. These arrays may use planar surfaces similar to DNA arrays, or use microspheres in suspension ("liquid arrays"). Also, many high quality autoantigens are now commercially available, including recombinant antigens. The vast amount of information that can be generated by measuring multiple antibodies for multiple patients has created demand for data processing. Software programs to aid physicians in reviewing multiple inputs as an aid to disease diagnosis and classification have been available for twenty years. Initial work used the "expert systems" approach; more recently pattern recognition has been widely evaluated because of the improvements in software programs and computational speed. The use of antibody data, generated in protein arrays, may assist in establishing diagnosis, in identifying potentially significant antibody patterns in advance of clinical symptoms, and in classifying patients based on expected disease progression. PMID- 16697964 TI - Immunoglobulin variable-region gene mutational lineage tree analysis: application to autoimmune diseases. AB - Lineage trees have frequently been drawn to illustrate diversification, via somatic hypermutation (SHM), of immunoglobulin variable-region (IGV) genes. In order to extract more information from IGV sequences, we developed a novel mathematical method for analyzing the graphical properties of IgV gene lineage trees, allowing quantification of the differences between the dynamics of SHM and antigen-driven selection in different lymphoid tissues, species, and disease situations. Here, we investigated trees generated from published IGV sequence data from B cell clones participating in autoimmune responses in patients with Myasthenia Gravis (MG), Rheumatoid Arthritis (RA), and Sjogren's Syndrome (SS). At present, as no standards exist for cell sampling and sequence extraction methods, data obtained by different research groups from two studies of the same disease often vary considerably. Nevertheless, based on comparisons of data groups within individual studies, we show here that lineage trees from different individual patients are often similar and can be grouped together, as can trees from two different tissues in the same patient, and even from IgG- and IgA expressing B cell clones. Additionally, lineage trees from most studies reflect the chronic character of autoimmune diseases. PMID- 16697965 TI - Recombinant antibodies with T-cell receptor-like specificity: novel tools to study MHC class I presentation. AB - MHC class I molecules play a central role in the immune response against a variety of cells that have undergone malignant transformation by shaping the T cell repertoire and by presenting peptide antigens from endogeneous antigens to CD8+ cytotoxic T cells. Because of their unique specificity such MHC-peptide complexes are a desirable target for novel approaches in immunotherapy. Targeted delivery of toxins or other cytotoxic drugs to cells which express specific MHC peptide complexes that are involved in the immune response against cancer or viral infections would allow for a specific immunotherapeutic treatment of these diseases. We have recently demonstrated that antibodies with the antigen specific, MHC restricted specificity of T cells can be generated by taking advantage of the selection power of phage display technology. In addition to their tumor targeting capabilities antibodies that mimic the fine specificity of T cell receptors can serve as valuable research reagents that enable to study human class I peptide-MHC ligand-presentation as well as TCR-peptide-MHC interactions. T-cell receptor-like antibody molecules may prove to be useful tools for studying MHC class I antigen presentation in health and disease as well as for therapeutic purposes in cancer, infectious diseases, and autoimmune disorders. PMID- 16697966 TI - Genome-wide SNP association: identification of susceptibility alleles for osteoarthritis. AB - The successful identification of genes involved in common human disorders is dependent upon availability of informative sample sets, validated marker panels, a high-throughput scoring technology, and a strategy for combining these resources. We have developed a universal platform based on mass spectrometry (MassARRAY) for analyzing nucleic acids with high precision and accuracy. To fuel this technology we have generated more than 100,000 validated assays for single nucleotide polymorphisms (SNPs) covering virtually all known and predicted human genes, and a large DNA sample bank from more than 50,000 consented diseased (case) and healthy (control) individuals. Taking advantage of MassARRAY's capability for quantitative analysis of nucleic acids, allele frequencies are estimated in sample pools containing large numbers of individual DNAs. Comparing frequencies between case and control pools as a first-pass filtering step is a tremendous advantage in throughput and cost over individual genotyping. We have employed this approach in numerous genome-wide association studies to identify genes implicated in common complex diseases, including osteoarthritis (OA). Access to additional patient samples through collaborations allows us to conduct replication studies that validate true disease genes. These discoveries will expand our understanding of genetic disease predisposition, and our capabilities for early diagnosis and improved therapeutic approaches. PMID- 16697967 TI - Molecular mechanisms of TNF-alpha-induced apoptosis in naive and memory T cell subsets. AB - Aging in humans is associated with progressive decline in T cell function, hyperimmunoglobulinemia, increased prevalence of autoantibodies and decline in naive CD8(+) T cells and accumulation of memory T cells, which appears to be oligoclonal and display feature of senescence, that is, decreased replication, short telomere length and resistance to apoptosis. Recently memory T cells have been further subdivided into central and effector memory T cells, based upon their migratory and homing properties. They are identified by a number of cell surface makers. In this brief review we will discuss molecular mechanisms of apoptosis in naive and various types of memory T cells to possibly explain the changes observed in aging, which are very similar to certain autoimmune diseases. PMID- 16697968 TI - Prevention of gonadal toxicity and preservation of gonadal function and fertility in young women with systemic lupus erythematosus treated by cyclophosphamide: the PREGO-Study. AB - BACKGROUND: With dramatically improved survival rates of SLE patients, comorbidity and long-term damage such as premature ovarian failure (POF) gain increasing importance. In the Erlangen cohort, 14% of cyclophosphamide treated patients younger than 41 years have POF, which is a common consequence of cyclophosphamide treatment. PATIENTS AND METHODS: We tested the concentrations of FSH and LH, before, during and after cyclophosphamide treatment in 63 premenopausal women with SLE without ovarian protection and initiated the PREGO Study (Prospective randomized study on protection against gonadal toxicity) in patients with SLE. RESULTS: In lupus patients treated with cyclophosphamide, 60% suffered from POF and hypergonadotropic amenorrhea. Whereas the POF rate was <50% in women below 30 years, it was 60% between 30 and 40 years. The cumulative dosage of cyclophosphamide also strongly influenced POF rate. CONCLUSIONS: Our present results, with a high POF rate in Cyclophosphamide treated SLE patients demonstrate the urgent need for ovarian protection in this patient group. Besides POF these women are at high risk for premature atherosclerosis which is the major cause of death in lupus. Following preliminary encouraging experience in women with lymphoma, in whom the temporary induction of a prepubertal hormonal milieu during chemotherapy, has significantly decreased the risk of POF, we have initiated the PREGO-Study, comparing randomised monthly injection versus no injection of gonadotropin-releasing hormone analogue (GnRH-a) to young SLE patients during cyclophosphamide therapy. PMID- 16697969 TI - The role of biologic therapies in the management of systemic vasculitis. AB - The recent development of biologic therapies capable of selectively targeting components of the immune system has revolutionised the treatment of inflammatory arthritides. The steady increase in use of biologic agents coupled with the expansion in the knowledge of the pathogenesis of vascular inflammation has led to their application in the treatment of primary systemic vasculitis. These agents may have a role in addition to or in place of conventional immunosuppression and also be effective when the latter fails to induce remission. The use of biologics as targeted therapies has also, in reverse, improved our understanding of the pathophysiology of vascular inflammation. While the advent of biologics heralds a new era in the management of the systemic vasculitis, evidence for their efficacy is still in its infancy and has yet to match that of conventional immunosuppressants. In this review, we examine the up to-date evidence for the use of biologics in systemic vasculitis, including TNF alpha inhibitors, and highlight the challenges facing their use. We examine the rationale for using biologics based on the pathophysiology of vasculitis. Issues of toxicity and pharmacovigilance with the use of biologics are also discussed. Finally, future directions and predictions are presented. PMID- 16697970 TI - Juvenile idiopathic arthritis. AB - One of the most enigmatic problems in rheumatology has been juvenile idiopathic arthritis (JIA). Firstly, the classification has often depended on clinical features that have variations between patients. Secondly, there are different classification schemes in usage and there are few objective serologic tests that help to resolve the differences between the criteria sets. Thirdly, only recently have significant advances been made in understanding the immunology and immunopathology of JIA and, in particular, new treatment options. In this review, we will define the historical basis of JIA and emphasize not only the clinical features, but also the immunological characteristics, the pathogenesis, and treatment options. We will also discuss, in particular, quality of life, psychosocial functioning, socioeconomic outcomes and the difficult area of mortality. Finally, this review will attempt to bridge genetic observations with clinical presentation. JIA represents a relatively common syndrome of pediatric onset rheumatologic disease and a better understanding of the clinical definition, the relationship to autoimmunity, and novel treatments with biologic agents are critical for improved patient care. PMID- 16697971 TI - Induction of early apoptosis and ROS-generation activity in human gingival fibroblasts (HGF) and human submandibular gland carcinoma (HSG) cells treated with curcumin. AB - OBJECTIVE: Curcumin [1] is well known to possess apoptosis-inducing activity in some cancer cells, but little is known about its activity in normal cells of oral origin, such as HGF. The aim of the present study was to clarify the relationship between early apoptosis in HGF and the induction of reactive oxygen species (ROS) generation by curcumin. DESIGN: We treated HGF and HSG cells with curcumin [1] and the related compounds biseugenol [2], eugenol [3], alpha-diisoeugenol [4], and isoeugenol [5] and measured cell survival (MTT method), ROS generation (DCFH DA staining), and induction of early apoptosis. Early apoptosis was detected by monitoring loss of mitochondrial membrane potential (DeltaPsi(m)) by JC-1 staining and externalization of phosphatidylserine (PS) on the cell surface by annexin V-FITC/PI staining combined with flow cytometry. RESULTS: The cytotoxic activities of curcumin [1] and [4] were similar and were nearly 10- to 100-fold stronger than those of the other compounds. Only curcumin was able to induce ROS generation and early apoptosis. Loss of DeltaPsi(m), PS externalization and ROS generation were significantly more pronounced in HGF cells than in HSG cells at curcumin concentrations lower than about 15microM, and were inhibited by the addition of the antioxidants N-acetyl-l-cysteine and glutathione. CONCLUSION: The potent PS externalization and loss of DeltaPsi(m) in curcumin-treated HGF cells appears to be mediated by ROS generation. PMID- 16697972 TI - The peripheral stalk of the mitochondrial ATP synthase. AB - The peripheral stalk of F-ATPases is an essential component of these enzymes. It extends from the membrane distal point of the F1 catalytic domain along the surface of the F1 domain with subunit a in the membrane domain. Then, it reaches down some 45 A to the membrane surface, and traverses the membrane, where it is associated with the a-subunit. Its role is to act as a stator to hold the catalytic alpha3beta3 subcomplex and the a-subunit static relative to the rotary element of the enzyme, which consists of the c-ring in the membrane and the attached central stalk. The central stalk extends up about 45 A from the membrane surface and then penetrates into the alpha3beta3 subcomplex along its central axis. The mitochondrial peripheral stalk is an assembly of single copies of the oligomycin sensitivity conferral protein (the OSCP) and subunits b, d and F6. In the F-ATPase in Escherichia coli, its composition is simpler, and it consists of a single copy of the delta-subunit with two copies of subunit b. In some bacteria and in chloroplasts, the two copies of subunit b are replaced by single copies of the related proteins b and b' (known as subunits I and II in chloroplasts). As summarized in this review, considerable progress has been made towards establishing the structure and biophysical properties of the peripheral stalk in both the mitochondrial and bacterial enzymes. However, key issues are unresolved, and so our understanding of the role of the peripheral stalk and the mechanism of synthesis of ATP are incomplete. PMID- 16697973 TI - Band 3 tyr-phosphorylation in human erythrocytes from non-pregnant and pregnant women. AB - Pregnancy is associated with changes in circulating red blood cells, mainly involving band 3 protein and membrane lipid peroxidation. Membrane band 3 is a multifunctional protein containing four Tyr-phosphorylatable residues which modulate the physiological status of erythrocytes by regulating glycolysis, cell shape and membrane transport. Erythrocytes from nine pregnant and 12 age-matched non-pregnant healthy women were subjected to oxidative and hyperosmotic stress conditions and the extent of band 3 Tyr-phosphorylation and membrane Syk recruitment as a membrane marker were evaluated. Results indicated that, in pregnancy, red blood cells show a decrease in band 3 Tyr-phosphorylation and a clear-cut rearrangement of band 3 protein within the membrane. In fact, band 3 shows a decrease in high molecular weight aggregates (HMWA), with different subdivision between Triton-soluble and -insoluble compartments, and an increase in proteolytic fragments. In conclusion, it is demonstrated that pregnancy is associated with membrane adjustments which reduce the sensitivity of erythrocytes to both oxidative and osmotic stress. Band 3 Tyr-phosphorylation is proposed as a new parameter in the evaluation of erythrocyte membrane arrangement. PMID- 16697974 TI - Infrared imaging microscopy of bone: illustrations from a mouse model of Fabry disease. AB - Bone is a complex tissue whose composition and properties vary with age, sex, diet, tissue type, health and disease. In this review, we demonstrate how infrared spectroscopy and infrared spectroscopic imaging can be applied to the study of these variations. A specific example of mice with Fabry disease (a lipid storage disease) is presented in which it is demonstrated that the bones of these young animals, while showing typical spatial variation in mineral content, mineral crystal size, and collagen maturity, do not differ from the bones of age- and sex-matched wild type animals. PMID- 16697975 TI - Peptide-membrane interactions and mechanisms of membrane destruction by amphipathic alpha-helical antimicrobial peptides. AB - Antimicrobial peptides (AMPs) have received considerable interest as a source of new antibiotics with the potential for treatment of multiple-drug resistant infections. An important class of AMPs is composed of linear, cationic peptides that form amphipathic alpha-helices. Among the most potent of these are the cecropins and synthetic peptides that are hybrids of cecropin and the bee venom peptide, mellitin. Both cecropins and cecropin-mellitin hybrids exist in solution as unstructured monomers, folding into predominantly alpha-helical structures upon membrane binding with their long helical axis parallel to the bilayer surface. Studies using model membranes have shown that these peptides intercalate into the lipid bilayer just below the level of the phospholipid glycerol backbone in a location that requires expansion of the outer leaflet of the bilayer, and evidence from a variety of experimental approaches indicates that expansion and thinning of the bilayer are common characteristics during the early stages of antimicrobial peptide-membrane interactions. Subsequent disruption of the membrane permeability barrier may occur by a variety of mechanisms, leading ultimately to loss of cytoplasmic membrane integrity and cell death. PMID- 16697976 TI - Multimodal assessment of disgust in contamination-related obsessive-compulsive disorder. AB - The present study utilizes multiple methods to examine the relationship between disgust and contamination-related obsessive-compulsive disorder (OCD) symptoms in an analogue sample. Questionnaire findings revealed that participants with high OCD contamination concerns showed stronger disgust sensitivity than did participants with low OCD contamination symptoms after controlling for negative affect. High OCD participants (N=30) also reported significantly more disgust than did low OCD participants (N=30) when exposed to a disgust-inducing video, whereas no significant between-group differences were detected on other negative emotional dimensions. Results from a series of disgust-specific behavioral avoidance tasks (BATs) revealed that high OCD participants demonstrated both less compliance and less approach behavior. Subsequent analysis also revealed that disgust sensitivity generally mediated avoidance on the BATs among high OCD subjects. High OCD participants also rated the BATs as more fearful and disgusting than did low OCD participants, with disgust generally emerging as the dominant emotional response. The results are consistent with a disgust-based, disease-avoidance approach in understanding contamination-related OCD themes. PMID- 16697977 TI - Cortical influences on rapid brainstem plasticity. AB - Cortical contributions to brainstem plasticity in the somatosensory system are poorly understood. Tactile receptive fields (RFs) of brainstem dorsal column nuclei (DCN) neurons rapidly enlarge when peripheral inputs are disrupted by local anesthetic blocks with lidocaine (LID). Cortical inputs appear to influence this plasticity because enlargements have been shown to be greater when cortical inputs are disrupted. Like disruptions of peripheral inputs, disruptions of DCN inhibition by DCN administration of the GABAA receptor antagonist bicuculline methiodide (BMI) also cause rapid enlargements of DCN RFs when cortical inputs are intact. These findings leave questions about interactions between cortical inputs, DCN inhibition, and DCN RF plasticity. To study potential interactions, the present experiments evaluated RF sizes of DCN tactilely responsive neurons in anesthetized rats following DCN microinjection of BMI when cortical inputs were acutely disrupted or intact. These tests were also supplemented by subsequent LID tests to directly compare post-BMI and post-LID effects on the same RF. BMI caused DCN RF enlargements when cortical inputs were disrupted or intact; however, enlargements after cortical input disruption were greater than when cortical inputs were intact. Following RF enlargement and retraction after BMI, LID often caused a second enlargement of the same RF, across skin that partially matched skin involved in the enlargement after BMI. This occurred when cortical inputs were disrupted or intact. We hypothesize that cortical inputs are not required for BMI and LID to initiate partially matching enlargements in individual DCN tactile RFs, however, cortical inputs constrain magnitudes of these enlargements. PMID- 16697978 TI - Soluble amyloid precursor protein alpha reduces neuronal injury and improves functional outcome following diffuse traumatic brain injury in rats. AB - Amyloid precursor protein (APP) has previously been shown to increase following traumatic brain injury (TBI). Whereas a number of investigators assume that increased APP may lead to the production of neurotoxic Abeta and be deleterious to outcome, the soluble alpha form of APP (sAPPalpha) is a product of the non amyloidogenic cleavage of amyloid precursor protein that has previously been shown in vitro to have many neuroprotective and neurotrophic functions. However, no study to date has addressed whether sAPPalpha may be neuroprotective in vivo. The present study examined the effects of in vivo, posttraumatic sAPPalpha administration on functional motor outcome, cellular apoptosis, and axonal injury following severe impact-acceleration TBI in rats. Intracerebroventricular administration of sAPPalpha at 30 min posttrauma significantly improved motor outcome compared to vehicle-treated controls as assessed using the rotarod task. Immunohistochemical analysis using antibodies directed toward caspase-3 showed that posttraumatic treatment with sAPPalpha significantly reduced the number of apoptotic neuronal perikarya within the hippocampal CA3 region and within the cortex 3 days after injury compared to vehicle-treated animals. Similarly, sAPPalpha-treated animals demonstrated a reduction in axonal injury within the corpus callosum at all time points, with the reduction being significant at both 3 and 7 days postinjury. Our results demonstrate that in vivo administration of sAPPalpha improves functional outcome and reduces neuronal cell loss and axonal injury following severe diffuse TBI in rats. Promotion of APP processing toward sAPPalpha may thus be a novel therapeutic strategy in the treatment of TBI. PMID- 16697979 TI - Distribution of neuropeptide W immunoreactivity and mRNA in adult rat brain. AB - Neuropeptide W (NPW) is a recently identified neuropeptide that binds to G protein-coupled receptor (GPR) 7, which is highly expressed in several discrete regions of the rodent brain including the central amygdaloid nucleus and bed nucleus of the stria terminalis. Although several reports suggested that NPW is implicated in the regulation of energy homeostasis and nociception, the precise physiological role of NPW has remained unclear. In this study, we examined distribution of NPW messenger RNA and NPW immunoreactivity in the adult rat brain. NPW-immunoreactive (ir) cells were detected in the ventral tegmental area, periaqueductal gray, and Edinger-Westphal nucleus. NPW-ir fibers were observed in several brain regions, including the lateral septum, bed nucleus of the stria terminalis, dorsomedial and posterior hypothalamus, central amygdaloid nucleus, CA1 field of hippocampus, interpeduncular nucleus, inferior colliculus, lateral parabrachial nucleus, facial nucleus, and hypoglossal nucleus. NPW-ir fibers were most abundantly observed in the central amygdaloid nucleus and the bed nucleus of the stria terminalis, which are regions implicated in fear and anxiety. These results suggest that NPW might be involved in the regulation of stress and emotive responses, especially in fear and anxiety-related physiological and behavioral functions. PMID- 16697980 TI - Treatment with deferoxamine increases neurons from neural stem/progenitor cells. AB - Neural transplantation is a promising approach for treating neurodegenerative disease. Neural stem/progenitor cells (NPCs) are self-renewing and multipotent and thus are good candidates for donor cells when they have been clearly defined to differentiate into neurons. As neuronal differentiation follows cell cycle exit, we investigated whether neuron production from NPCs is increased by treatment with cell cycle blockers. NPCs from E12.5 rat ventral mesencephalon were cultured as neurospheres in DMEM/F12 medium containing N2 supplements and bFGF. Treatment of NPCs with deferoxamine, a G1/S phase blocker, increased the number of beta-tubulin III-positive cells after differentiation, concomitant with increases of MAP2 mRNA and protein, and a decrease of GFAP protein. Further, an increase in beta-tubulin III/BrdU double-positive cells and a decrease in GFAP/BrdU double-positive cells were confirmed. In real-time PCR, the expressions of p21(cip1), p27(kip1) and p57(kip2) mRNAs remained unaltered for 8 h after treatment with deferoxamine but were significantly elevated after 1 day. Deferoxamine specifically enhanced the elevation of p27(kip1) mRNA at 1-2 days and the accumulation of p27(kip1) protein at 3 days, along with the activation of neuroD promoter and the elevation of neuroD mRNA. Transfection of p27(kip1) into NPCs induced activation of neuroD promoter and increase of number of beta-tubulin III-positive cells. These data suggest that pretreatment with deferoxamine increases the number of neurons from NPCs related to prolonged p27(kip1) elevation and activation of the neuroD signaling pathway. In this way, regulation of the cell cycle should be a useful first step in engineering NPCs for neural transplantation. PMID- 16697981 TI - Parasite glycoconjugates. Part 16: Synthesis of a disaccharide and phosphorylated di- and tri-saccharides from Leishmania lipophosphoglycan. AB - A neutral disaccharide beta-D-Galp-(1-->4)-alpha-D-Manp and phosphorylated di- and tri-saccharides beta-D-Galp-(1-->3)-[H(2)PO(3)-6]-beta-D-Galp O[CH(2)](8)CHCH(2) and beta-D-Galp-(1-->3)-[H(2)PO(3)-6]-beta-D-Galp-(1-->4) alpha-D-Manp, which are fragments of the phosphoglycan portion of the surface lipophosphoglycan from Leishmania donovani (the disaccharide) or Leishmania major (all three compounds), were prepared and used as TLC standards to help the identification and differentiation of the elongating and branching beta-D galactosyl transferase activities in Leishmania. The phosphosaccharides were synthesised using the H-phosphonate method for phosphorylation. PMID- 16697982 TI - Improved synthesis of dicyclohexylidene protected quebrachitol and its use in the synthesis of L-chiro-inositol derivatives. AB - A modified synthesis of 1L-1,2:3,4-di-O-cyclohexylidene-5-O-methyl-chiro-inositol has been accomplished that improves the overall procedure, yield, and environmental aspects of its formation. Several inositol analogues have been prepared from this intermediate for testing as biosynthetic inhibitors of glycosyl-phosphatidylinositol (GPI) anchor formation. PMID- 16697983 TI - Synthesis of a 3'-naphthamido-LacNAc fluorescein conjugate with high selectivity and affinity for galectin-3. AB - Described is the synthesis of a fluorescent LacNAc derivative appended with a 3' deoxy-3'-naphthamido functionality, 2-(fluorescein-5/6-amido)ethyl 3-deoxy-3-(2 naphthamido)-beta-D-galactopyranosyl-(1-->4)-2-acetamido-2-deoxy-beta-D glucopyranoside, which confers high affinity (Kd 170 nM) and selectivity for galectin-3 via a stacking interaction with Arg144. Its use as a selective and sensitive galectin-3 probe is demonstrated with fluorescence polarization measurements. PMID- 16697984 TI - Neutral pentosides surfactants issued from the butadiene telomerization with pentoses: preparation and amphiphilic properties. AB - Interfacial properties of octadienyl pentosides prepared by the palladium catalyzed telomerization of butadiene with free pentoses have been evaluated and compared to those of mixtures issued from the autoclaving process. PMID- 16697985 TI - Synthesis of a novel glycosphingolipid from the millipede, Parafontaria laminata armigera, and the assembly of its carbohydrate moiety into multivalent structures. AB - A novel glycosphingolipid, beta-D-Manp-(1-->4)-[alpha-L-Fucp-(1-->3)]-beta-D-Glcp (1-->1)-Cer, found in the millipede, Parafontaria laminata armigera, and multivalent derivatives of its carbohydrate moiety were synthesized. As the key step, the target glycolipid (1) was obtained through an inversion reaction at the 2-position of a beta-glucopyranoside residue yielding a beta-mannopyranoside. In addition, the synthesis of fluorescently labeled trimer and tetramer glycoconjugates (2, 3) was achieved by iterative amide bond formation using a monomer unit (24). PMID- 16697986 TI - Glyoxal formation by Fenton-induced degradation of carbohydrates and related compounds. AB - In this paper, we provide a systematic analysis of glyoxal (1) formation from a range of monosaccharides and related compounds, to determine their potential role as sources of this alpha-oxoaldehyde in vivo. Substrates were reacted with the Fenton reagent (Fe(2+)/EDTA/H(2)O(2)) and the mixtures were analyzed by HPLC using the 6-hydroxy-2,4,5-triaminopyrimidine fluorimetric assay. The rank order of hexoses and their derivatives as glyoxal sources was found to be fructose > glucose = mannose = galactose > glucose-6-phosphate > mannitol. Within the pentose group, arabinose and ribose gave the higher yields of 1 followed by deoxyribose and its adenine N-glycosides and ribulose. Among the tested substrates, three-carbon compounds, that is, trioses and glycerol, but not glyceraldehyde-3-phosphate, were by far the most effective sources of 1. The effects of H(2)O(2) and Fe(2+)/EDTA concentrations as well as of other metal ions were also investigated. PMID- 16697987 TI - DBU assisted expeditious synthesis of glycosyl dienes via glycosylated beta hydroxy esters. AB - DBU catalyzed condensation of 3-O-benzyl(methyl)-5,6-dideoxy-1,2-O-isopropylidene beta-L-threo-hept-4-enofuranuronates with different aldehydes produces the corresponding 3-O-benzyl(methyl)-6-carbethoxy-5,6-dideoxy-1,2-O-isopropylidene-7 phenyl-beta-L-threo-hept-4-enofuranoses. The latter on treatment with methanesulfonyl chloride followed by DBU catalyzed E2 reaction of the methanesulfonyloxy intermediates gave the respective 3-O-benzyl(methyl)-6 carbethoxy-5,6,7-trideoxy-1,2-O-isopropylidene-7-phenyl-beta-L-threo-hept-4,6 dienofuranose in moderate to good yields. PMID- 16697988 TI - Synthesis of multivalent lactose derivatives by 1,3-dipolar cycloadditions: selective galectin-1 inhibition. AB - Acetylene derivatives of phenylalanine, phenethylamine and the multifunctional unnatural amino acids, phenyl-bis-alanine and phenyl-tris-alanine, were synthesized and functionalized with 2-azidoethyl beta-D-galactopyranosyl-(1-->4) beta-D-glucopyranoside via regioselective copper(I)-mediated 1,3-dipolar cycloaddition to give a panel of mono-, di- and trivalent lactoside derivatives. Evaluation of the compounds as inhibitors against the tumour- and inflammation related galectin-1, -3, -4N, -4C, -4, -7, -8N and -9N revealed a divalent compound with a Kd value as low as 3.2 microM for galectin-1, which corresponded to a relative potency of 30 per lactose unit as compared to the natural disaccharide ligand lactose. This divalent compound had at least one order of magnitude higher affinity for galectin-1 than for any of the other galectins investigated. PMID- 16697989 TI - A novel oligoalginate lyase from abalone, Haliotis discus hannai, that releases disaccharide from alginate polymer in an exolytic manner. AB - We previously reported the isolation and cDNA cloning of an endolytic alginate lyase, HdAly, from abalone Haliotis discus hannai [Carbohydr. Res.2003, 338, 2841 2852]. Although HdAly preferentially degraded mannuronate-rich substrates, it was incapable of degrading unsaturated oligomannuronates smaller than tetrasaccharide. In the present study, we used conventional chromatographic techniques to isolate a novel unsaturated-trisaccharide-degrading enzyme, named HdAlex, from the digestive fluid of the abalone. The HdAlex showed a molecular weight of 32,000 on SDS-PAGE and could degrade not only unsaturated trisaccharide but also alginate and mannuronate-rich polymers at an optimal pH and temperature of 7.1 and 42 degrees C, respectively. Upon digestion of alginate polymer, HdAlex decreased the viscosity of the alginate at a slower rate than did HdAly, producing only unsaturated disaccharide without any intermediate oligosaccharides. These results indicate that HdAlex degrades the alginate polymer in an exolytic manner. Because HdAlex split saturated trisaccharide producing unsaturated disaccharide, we considered that this enzyme cleaved the alginate at the second glycoside linkage from the reducing terminus. The primary structure of HdAlex was deduced with cDNAs amplified from an abalone hepatopancreas cDNA library by the polymerase chain reaction. The translational region of 822 bp in the total 887-bp sequence of HdAlex cDNA encoded an amino acid sequence of 273 residues. The N-terminal sequence of 16 residues, excluding the initiation methionine, was regarded as the signal peptide of this enzyme. The amino-acid sequence of the remaining 256 residues shared 62-67% identities with those of the polysaccharide lyase family-14 (PL14) enzymes such as HdAly and turban-shell alginate lyase SP2. To our knowledge, HdAlex is the first exolytic oligoalginate lyase belonging to PL14. PMID- 16697990 TI - Synthesis of asparagine-linked bacillosamine. AB - Various types of protein glycosylation have been identified from prokaryotes. Recent investigations have revealed the presence of N-linked glycoproteins in the pathogenic bacterium, Campylobacter jejuni. The structure of this glycan is unique, consisting of 5 GalNAc and 1 Glc, in addition to 2,4-diacetamido-2,4,6 trideoxy-d-glucopyranose (bacillosamine; Bac), which is N-glycosidically linked to the side chain of asparagine (Asn). We synthesized Bac from a 2-azido-2-deoxy D-galactose derivative, which was further converted to the Asn-linked form. PMID- 16697991 TI - Structural determination of the O-antigenic polysaccharide from the Shiga toxin producing Escherichia coli O171. AB - The structure of the O-antigenic part of the lipopolysaccharide (LPS) obtained from the verotoxin-producing Escherichia coli O171 has been determined. (1)H and (13)C NMR spectroscopy techniques in combination with component analysis were used to elucidate the O-antigen structure of O-deacylated LPS. Subsequent NMR analysis of the native LPS revealed acetylation at O-7/O-9 of the sialic acid residue. The sequence of sugars was determined by inter-residue correlations in (1)H,(1)H-NOESY and (1)H,(13)C-heteronuclear multiple-bond correlation spectra. The O-antigen is composed of pentasaccharide repeating units with one equivalent of O-acetyl groups distributed over two positions: -->4)-alpha-Neu5Ac7,9Ac-(2- >6)-beta-D-Galp-(1-->6)-beta-DGlcp-->(1-->3)-beta-D-Galp-(1-->3)-beta-D-GalpNAc (1--> Based on biosynthetic considerations, this should also be the biological repeating unit. PMID- 16697992 TI - Concentration measurements of sucrose and sugar surfactants solutions by using the 1H NMR ERETIC method. AB - The ERETIC method has been used to determine precise concentrations of aqueous solutions of sucrose and sugar surfactants, namely octyl glucoside and fatty acid sucrose esters by (1)H NMR spectroscopy. The effects of NMR tuning, acquisition parameters, and spectrum processing on the measurement have been assessed in these particular cases. The linearity upholds over the whole concentration range, with both sucrose and octyl glucoside, whatever the physicochemical phenomena occurring, either an increasing viscosity or the micellization of the surfactant. For sucrose solutions, an accuracy of 2% is measured for concentrations between 0.1 and 200 mmol/L, which is consistent with literature data. PMID- 16697993 TI - Structural studies of the O-antigenic oligosaccharide from Vibrio salmonicida strain C2 isolated from Atlantic cod, Gadus morhua L. AB - We report the chemical structure of the oligosaccharide part of Vibrio salmonicida lipopolysaccharide serotype C2, isolated from Atlantic cod (Gadus morhua L.). The structure was established by NMR spectroscopy and mass spectrometry. It is concluded that the oligosaccharide has the following structure in which L-alpha-D-Hepp is L-glycero-alpha-D-manno-heptopyranose, D alpha-D-Hepp is D-glycero-alpha-D-manno-heptopyranose, alpha-NonA is 5 acetamidino-7-acetamido-3,5,7,9-tetradeoxy-L-glycero-alpha-D-galacto-nonulosonic acid, and PEA is phosphoethanolamine. [chemical structure: see text] PMID- 16697994 TI - A new synthesis of alpha-arbutin via Lewis acid catalyzed selective glycosylation of tetra-O-benzyl-alpha-D-glucopyranosyl trichloroacetimidate with hydroquinone. AB - alpha-Arbutin has huge application potentials in the cosmetic industry, as its inhibitory effect on human tyrosinase is stronger than that of its naturally occurring anomer arbutin (4-hydroxyphenyl beta-D-glucopyranoside). Enzymatic synthesis was preferred for alpha-arbutin previously, and now a new chemical synthesis is reported. The reaction of tetra-O-benzyl-alpha-D-glucopyranosyl trichloroacetimidate, as glycosyl donor, with hydroquinone was initiated by catalytic amounts of trimethylsilyl trifluoromethanesulfonate (TMSOTf), resulting in 4-hydroxyphenyl 2,3,4,6-tetra-O-benzyl-alpha-D-glucopyranoside with high stereoselectivity and yield, and then to alpha-arbutin quantitatively after deprotection. PMID- 16697995 TI - O-Isopropylidenation of carbohydrates catalyzed by vanadyl triflate. AB - Vanadyl triflate has been identified as a mild and efficient catalyst for the chemoselective O-isopropylidenation of functionalized carbohydrates with acetone and acetone equivalents. The current protocol is compatible with a diverse array of protecting groups and the products can be readily isolated by simple aqueous wash. PMID- 16697996 TI - Structure of the O-polysaccharide of Proteus mirabilis CCUG 10705 (OF) containing an amide of D-galacturonic acid with L-alanine. AB - The structure of the O-polysaccharide of Proteus mirabilis CCUG 10705 (OF) was determined by chemical analyses along with one- and two-dimensional (1)H and (13)C NMR spectroscopy. The polysaccharide was found to contain an amide of D galacturonic acid with L-alanine and based on the uniqueness of the O polysaccharide structure and serological data, it was suggested to classify P. mirabilis OF into a new separate Proteus serogroup, O74. A weak cross-reactivity of P. mirabilis OF and P. mirabilis O5 was observed and accounted for by a similarity of their O-repeating units. The following structure of the polysaccharide of P. mirabilis OF was established: [chemical structure: see text] PMID- 16697997 TI - Feruloyl esterases as a tool for the release of phenolic compounds from agro industrial by-products. AB - Agro-industrial by-products are a potential source of added-value phenolic acids with promising applications in the food and pharmaceutical industries. Here two purified feruloyl esterases from Aspergillus niger, FAEA and FAEB were tested for their ability to release phenolic acids such as caffeic acid, p-coumaric acid and ferulic acid from coffee pulp, apple marc and wheat straw. Their hydrolysis activity was evaluated and compared with their action on maize bran and sugar beet pulp. The specificity of both enzymes against natural and synthetic substrates was evaluated; particular attention was paid to quinic esters and lignin monomers. The efficiency of both enzymes on model substrates was studied. We show the ability of these enzymes to hydrolyze quinic esters and ester linkages between phenolic acids and lignin monomer. PMID- 16697998 TI - Synthesis of the sugar moiety of TIME-EA4, a glycopeptide isolated from silkworm diapause eggs. AB - We describe the efficient synthesis of the tetrasaccharide, 2-O-acetyl-3,4,6-tri O-benzyl-alpha-D-mannopyranosyl-(1-->6)-2,4-di-O-acetyl-3-O-allyl-beta-D mannopyranosyl-(1-->4)-3,6-di-O-benzyl-2-deoxy-2-phthalimido-beta-D glucopyranosyl-(1-->4)-3,6-di-O-benzyl-2-deoxy-2-phthalimido-beta-D glucopyranosyl azide, which is the protected form of the sugar unit of TIME-EA4 that is isolated from the diapausing eggs of the silkworm, Bombyx mori. The beta linked D-mannoside of the tetrasaccharide was obtained using the conventional oxidation-reduction method for inversion of the configuration at the C-2 hydroxyl group of beta-D-glucoside. The reduction was effected with NaBH(4) in a methanolic solution in a ratio of 98:2 in favor of the beta-D-mannoside that was obtained in 87% yield. PMID- 16697999 TI - Contrasting reactivity of thioglucoside and selenoglucoside donors towards promoters: implications for glycosylation stereocontrol. AB - The stereochemical outcome of glycosylation reactions with model thioglycosides and selenoglycosides proved to be dependent on the source of promoter iodonium ion, with iodine giving different results to N-iodosuccinimide (NIS) alone or N iodosuccinimide/trimethylsilyltrifluoromethanesulfonate (NIS/TMSOTf). In contrast to armed thioglycosides, which anomerise, and disarmed thioglycosides, which do not react, both armed and disarmed selenoglycosides give rise to the corresponding glycosyl iodides when reacted with iodine. Further, whilst the single electron transfer agent DDQ alone is an ineffective promoter, in combination with iodine it produces better acetonitrile-assisted beta stereoselectivity with both thioglycosides and selenoglycosides than does tris(4 bromophenyl)aminium hexachloroantimonate (BAHA). PMID- 16698000 TI - A de novo approach to C-branched inositols: synthesis of a myo-inositol precursor for C-linked glycosyl phosphatidylinositols. AB - C-Linked glycosyl inositols are valuable structure-activity probes because of their greater hydrolytic stability and different conformational behavior compared with their parent O-glycosides. Simple C-branched inositols are synthetic precursors to these and other groups of inositol mimetics. Herein is described a de novo synthesis of C-branched inositols that contain a versatile ethenyl side chain for elaboration into more complex appendages. The approach centers on a stereoselective oxocarbenium ion-allylsilane cyclization and provides C-branched inositols with different stereochemical motifs. The synthesis of C-ethenyl-di-O isopropylidene-myo-, neo-, epi-, and allo-inositols is discussed. PMID- 16698001 TI - Association of RhoGDIalpha with Rac1 GTPase mediates free radical production during myocardial hypertrophy. AB - OBJECTIVE: Reactive oxygen species (ROS) contribute to the pathogenesis of myocardial hypertrophy. NADPH oxidase is a major source of ROS production. The small GTPase Rac1 mediates the activation of NADPH oxidase; however, the mechanism of Rac1 activation is incompletely understood. METHODS AND RESULTS: Transaortic constriction (TAC, C57/Bl6 mice, 360 microm, 21 days) increased the ratio of heart to body weight from [ per thousand] SHAM 4.16+/-0.09 to TAC 7.1+/ 0.37, p<0.01. Treatment with rosuvastatin prevented pressure-induced cardiac hypertrophy (5.5+/-0.18, p<0.05). TAC induced a 4-fold up-regulation of myocardial NADPH oxidase activity as well as Rac1 activity; both effects were absent in statin-treated animals. In cultured rat cardiomyocytes, treatment with angiotensin II (AngII) increased translocation of Rac1 to cell membranes and Rac1 activity. AngII altered neither expression nor tyrosine phosphorylation of GTPase activating protein GAP-p190 and the guanine nucleotide exchange factors Vav and Tiam. Transaortic constriction as well as AngII increased the binding of Rho guanine nucleotide dissociation inhibitor (RhoGDIalpha) to Rac1. The association of RhoGDIalpha with Rac1 was mediated by phosphatidylinositol 3-kinase and depended on geranylgeranylation. Statin treatment inhibited RhoGDIalpha-Rac1 binding both in cultured cardiomyocytes and during myocardial hypertrophy in vivo. Transfection with RhoGDIalpha siRNA constructs potently reduced RhoGDIalpha protein expression, decreased AngII-induced superoxide production and lipid peroxidation, and inhibited AngII-induced leucine incorporation. CONCLUSIONS: Myocardial hypertrophy is characterized by activation of Rac1 and NADPH oxidase. The association of the regulatory protein RhoGDIalpha with Rac1 represents a necessary step in the Rac1-dependent release of ROS. Rac1-RhoGDIalpha binding may represent a target for anti-hypertrophic pharmacologic interventions, potentially by statin treatment. PMID- 16698002 TI - Morphine modulates monocyte-macrophage conversion phase. AB - Monocyte migration and their activation into the macrophage phenotype play a role in the modulation of tissue injury. We studied the effect of morphine on the monocyte-macrophage conversion phase (MMCP). Phorbol 12-myristate 13-acetate (PMA) activated THP-1 cells and promoted their adhesion to the substrate. Morphine inhibited PMA-induced MMCP. However, opiate receptor antagonists attenuated this effect of morphine. Interestingly, PMA as well as morphine stimulated superoxide production by monocytes. Superoxide dismutase (SOD) not only inhibited PMA-mediated MMCP but also attenuated the inhibitory effect of morphine. PMA not only enhanced adhesion of monocytes to a filter but also promoted their migration. These findings suggest that the PMA-induced macrophage phenotype conversion may be accelerating their migration; whereas, morphine may be preventing the migration of monocytes by inhibiting MMCP. PMID- 16698003 TI - Some insights into the mode of action of butadiene by examining the genotoxicity of its metabolites. AB - 1,3-Butadiene (BTD) is an important commodity chemical and air pollutant that has been shown to be a potent carcinogen in mice, and to a lesser extent, a carcinogen in rats. To better assess butadiene's carcinogenic risk to humans, it is important to understand its mode of action and how this relates to differences in responses among species. In a series of in vitro experiments, lymphocytes from rats, mice, and humans were exposed to 3,4-epoxy-1-butene (EB) or 1,2:3,4 diepoxybutane (DEB) for 1h at the G(0) stage of the cell cycle, stimulated to divide, and cultured to assess the ability of these metabolites to induce sister chromatid exchange (SCE) and chromosome aberrations (CAs). EB induced no increases in SCEs or CAs in the cells from the three species. DEB was a potent SCE- and CA-inducer, with the results being similar in each rodent species. The response for SCEs seen in the human cells was more complex, with genetic polymorphism for glutathione-S-transferases (GST) possibly modulating the response. The single cell gel electrophoresis assay was used on genetically engineered V79 cell lines to investigate a possible influence of GST status. Experiments were also conducted to investigate the reason for EB's failure to induce SCEs or CAs in G(0) cells. The results indicate that EB-induced DNA damage was repaired before DNA synthesis in unstimulated lymphocytes, but EB caused a large increase in SCEs if actively cycling cells were treated. Thus, the results indicate that DEB damage is persistent in G(0) cells, and DEB is a much more potent genotoxicant than EB. The carcinogenic effect of butadiene will most likely depend on the degree to which DEB is produced and reaches target tissues, and to a lesser extent on the ability of EB to reach actively dividing or repair deficient cells. PMID- 16698004 TI - Determination of carbohydrate-deficient transferrin in human serum using the Bio Rad %CDT by HPLC test. AB - BACKGROUND: Carbohydrate-deficient transferrin (CDT) in serum is a biomarker used to identify individuals with sustained, heavy alcohol consumption. This study evaluated the performance of a new commercial method for CDT, the Bio-Rad %CDT by HPLC test, that measures relative amounts of separate transferrin glycoforms in proportion to total transferrin. METHOD: The samples used were two human serum pools (low/high disialotransferrin), 150 clinical sera with low to highly elevated disialotransferrin values, and 18 genetic transferrin variants. Glycoforms are separated on a gradient HPLC system, followed by specific measurement of the iron-transferrin complex at 460 nm. Comparison was made with an HPLC candidate reference method on an Agilent 1100 LC system. RESULTS: The Bio Rad %CDT by HPLC test allowed for reproducible separation and quantification of the transferrin glycoforms within approximately 6 min. Genetic variants were readily identified. For the low and high serum pool, the total CV was 8.5% and 4.3%, respectively. The relative amounts of disialotransferrin, the main CDT glycoform, were in good agreement with the results of the HPLC candidate reference method (r(2)=0.998, p<0.0001). CONCLUSIONS: The present results demonstrated that the Bio-Rad %CDT by HPLC test is appropriate for confirmatory and routine %CDT testing in human serum, with the added advantage over previously published HPLC methods of an improved serum sample pretreatment and a shorter total analysis time. PMID- 16698005 TI - The exciting story of cardiac biomarkers: from retrospective detection to gold diagnostic standard for acute myocardial infarction and more. AB - This paper reviews the history of the contribution of the laboratory medicine to clinical cardiology and discusses the most important steps in this field. Until 20 years ago, the clinical laboratory only placed at the cardiologist's disposal a few assays for the retrospective detection of cardiac tissue necrosis, such as enzymatic methods for creatine kinase and lactate dehydrogenase activities. However, in the latter part of the 20th century, highly sensitive and specific assays, such as cardiac troponins, as well as assays for markers of myocardial function, such as cardiac natriuretic peptides, rapidly changed the scenario of clinical management of patients with cardiac diseases, assigning to the laboratory a pivotal role in the overall diagnostic flow. This is witnessed by the recent incorporation of these markers into international guidelines and in the redefinition of myocardial infarction. For the foreseeable future, new serum markers of myocardial ischemic, i.e. reversible, injury or related to coronary plaque instability and disruption are expected. PMID- 16698006 TI - Novel mutation, c.1234delA, in the DAX1 gene in congenital adrenal hypoplasia. PMID- 16698007 TI - The effect of aspirin and vitamins C and E on HbA1c assays. AB - BACKGROUND: Aspirin (ASA) and vitamins C and E may inhibit non-enzymatic glycation in vivo and may also interfere with HbA(1c) assays, masking true results. We investigated the effect of usual doses of ASA, vitamin C and E on HbA1c levels in a group of non-diabetic volunteers. METHODS: A randomized clinical trial was performed with 28 healthy non-diabetic individuals. Subjects were allocated to take ASA 200 mg/day, vitamin C 1 g/day, vitamin E 400 mg/day, or to a control group, for a period of 4 months. Blood samples were collected at baseline and at monthly intervals for HbA1c analysis by HPLC Variant II (BioRad), HPLC L-9100 (Merck - Hitachi) and Tina Quant HbA(1c) II immunoassay (Roche). RESULTS: HbA(1c) levels of the control, vitamin C and E groups did not change throughout the study, independently of the method used. HbA(1c) measured by Hitachi L-9100 HPLC increased significantly (P=0.033) at 4 months after ASA intake, although this increase was of only 0.17%. CONCLUSIONS: Treatment with vitamins C and E in pharmacological doses does not have any impact on HbA1c measurements in non-diabetic patients with the three methods employed. ASA induces a modest, not clinically relevant, increase in HbA1c levels with one of the methods. PMID- 16698008 TI - Plasma heart-type fatty acid binding protein is superior to troponin and myoglobin for rapid risk stratification in acute pulmonary embolism. AB - BACKGROUND: Irreversible right ventricular (RV) failure with myocardial damage may precipitate fatal outcome in acute pulmonary embolism (APE). Cytoplasmic heart-type fatty acid binding protein (H-FABP) is a sensitive and specific biomarker of myocardial damage. We assessed which biomarker of myocardial damage or RV stretching is the most useful for short-term risk stratification in APE. METHODS: We analyzed 77 patients (51 F, 26 M) aged 65.3+/-16.0 years with confirmed APE. On admission, systemic blood pressure and transthoracic echocardiography (for RV overload) were recorded and plasma concentrations of myoglobin (Mb), cardiac troponin T (cTnT), N-terminal fragment of proBNP (NT proBNP) and H-FABP were evaluated. RESULTS: Fifteen (19.5%) patients died and 24 (31.2%) experienced complicated clinical course (CCC) death/thrombolysis/cardiopulmonary resuscitation/intravenous vasopressors. Hazard ratio analysis demonstrated that plasma H-FABP, Mb, cTnT and NT-proBNP concentrations predicted fatal outcome. When only APE-related deaths were considered, plasma H-FABP concentrations indicated fatal outcome. Multivariate hazard ratio analysis revealed H-FABP as the only 30-day mortality predictor (HR 1.02 CI 95% 1.01-1.05). CONCLUSIONS: H-FABP measured on admission is useful for short-term risk stratification in APE. It appears to be superior to cTnT, NT proBNP and Mb in the prediction of 30-day APE-related mortality. PMID- 16698009 TI - Cryopreservation of Crassostrea gigas vesicular cells: viability and metabolic activity. AB - Cryopreservation is widely used for long-term conservation of various tissues, embryos or gametes. However, few studies have described cryopreservation of invertebrate primary cell cultures and more particularly of marine invertebrate somatic cells. This technique would however be of great interest to facilitate the study of various metabolic processes which vary seasonally. The aim of the present study was to develop a protocol for cryopreservation of Crassostrea gigas vesicular cells. Different parameters were adjusted to improve recovery of cells after freezing. The most efficient cryoprotectant agent was a mix of Me(2)SO, glycerol, and ethylene glycol (4% each). The optimal cooling rate was -1 degrees Cmin(-1) down to -70 degrees C before transfer into liquid nitrogen. In these conditions the percentage of viable cells reached 70% of the control. The glucose metabolism of thawed cells was evaluated using radioactive glucose as a tracer. Immediately after thawing, glucose uptake involving membrane transporters was greatly reduced (24% of control) whereas glucose incorporation into glycogen was less affected (68% of control). PMID- 16698010 TI - Fertilization triggers localized activation of Src-family protein kinases in the zebrafish egg. AB - Fertilization triggers activation of Src-family kinases in eggs of various species including marine invertebrates and lower vertebrates. While immunofluorescence studies have localized Src-family kinases to the plasma membrane or cortical cytoplasm, no information is available regarding the extent to which these kinases are activated in different regions of the zygote. The objective of the present study was to detect the subcellular distribution of activated Src-family kinases in the fertilized zebrafish egg. An antibody specific for the active, non-phosphorylated form of Src-family PTKs was used to detect these activated kinases by immunofluorescence. The results demonstrate that Fyn, and possibly other Src family members are activated by dephosphorylation of the C-terminal tyrosine at fertilization. The activated Src family kinases are asymmetrically distributed around the egg cortex with an area of higher kinase activity localized adjacent to the micropyle near the presumptive animal pole. Fertilization initially caused elevation of kinase activity in the cytoplasm underlying the micropyle, but this quickly spread to involve the entire zygote cortex. Later, during egg activation, formation of the blastodisc involved concentration of active Src-family kinase in the blastodisc cortex. As cytokinesis began, activated Src-family kinases were no longer limited to the cortex, but became more evenly distributed in the clear apical cytoplasm of the blastomeres. The results demonstrate that the cortex of the zebrafish egg is functionally differentiated and that fertilization triggers localized activation of Src-family kinases at the point of sperm entry, which subsequently progresses through the entire egg cortex. PMID- 16698011 TI - Central nervous system depressant action of flavonoid glycosides. AB - The pharmacological effects on the central nervous system (CNS) of a range of available flavonoid glycosides were explored and compared to those of the glycosides 2S-hesperidin and linarin, recently isolated from valeriana. The glycosides 2S-neohesperidin, 2S-naringin, diosmin, gossipyn and rutin exerted a depressant action on the CNS of mice following i.p. injection, similar to that found with 2S-hesperidin and linarin. We demonstrate in this work that these behavioural actions, as measured in the hole board, thiopental induced sleeping time and locomotor activity tests, are unlikely to involve a direct action on gamma-aminobutyric acid type A (GABA(A)) receptors. The corresponding aglycones were inactive, pointing to the importance of the sugar moieties in the glycosides in their CNS depressant action following systemic administration. The pharmacological properties of the flavonoid glycosides studied here, in addition to our previous results with hesperidin and linarin, opens a promising new avenue of research in the field. PMID- 16698012 TI - Anti-inflammatory activity of azithromycin attenuates the effects of lipopolysaccharide administration in mice. AB - Macrolide antibacterials inhibit the production of various cytokines and the migration of inflammatory cells. These anti-inflammatory actions of macrolides may be beneficial in attenuating inflammatory processes involved in bacterial sepsis. Therefore, we investigated the ability of azithromycin to attenuate the deleterious effects of lipopolysaccharide (LPS), in three different LPS-induced inflammatory models. Our results show that azithromycin (10 and 100 mg/kg) significantly attenuated the intraperitoneal LPS-induced increase in plasma TNF alpha concentration. It also increased survival rate in a septic shock model in mice challenged with intravenous LPS. Oral treatment with azithromycin (up to 300 mg/kg) was less effective in suppressing neutrophil infiltration into the lungs 24 h after intranasal LPS challenge, possibly because of a slower onset of action or inadequate dosing. In the same model, azithromycin given intraperitoneally significantly improved inflammatory markers (total cell number, neutrophil percentage and MIP-2 concentration) in bronchoalveolar lavage fluid. In conclusion, azithromycin exhibits significant anti-inflammatory properties but the potency of such effects varies depending on the experimental model and route of administration. PMID- 16698013 TI - Anti-inflammatory effect of jeongshintang through suppression of p38 activation in human astrocytoma, U373MG cells. AB - Jeongshintang (JST) is a Korean herbal prescription, which has been successfully used for cerebral diseases. However, the anti-inflammatory effect of JST on Alzheimer's disease (AD) is still not fully understood. In this study, we investigated the effects of JST in attenuating the inflammatory response induced by interleukin (IL)-1beta plus beta-amyloid [1-42] fragment (A beta) in the human astrocyte cell line, U373MG. The production of IL-6, IL-8, and prostaglandin (PG)E2 was significantly increased by IL-1beta plus A beta (1-42) in a time dependent manner (P < 0.05). JST significantly inhibited the IL-1beta plus A beta (1-42)-induced IL-6, IL-8, and PGE2 production at 24 h (P < 0.05). Maximal inhibition rate of IL-6, IL-8, and PGE2 production by JST was about 54.40%, 56.01%, and 44.06% respectively. JST (0.01-1 mg/ml) also attenuated the expression of cyclooxygenase (COX)-2 and activation of p38 MAPK induced by IL 1beta and A beta (1-42). These results demonstrated that JST has an anti inflammatory effect, which might explain its beneficial effect in the treatment of various neurodegenerative diseases such as AD. PMID- 16698014 TI - Structural model of the OPA1 GTPase domain may explain the molecular consequences of a novel mutation in a family with autosomal dominant optic atrophy. AB - Autosomal dominant optic atrophy (ADOA) is the most frequent hereditary optic neuropathy. Three loci have been reported for ADOA: a major locus, harboring all identified mutations to date, maps to 3q28 (OPA1), a second locus is linked to 18q12.2-q12.3 (OPA4) and a third locus on 22q12.1-q13.1 (OPA5) has been reported recently. We describe a six-generation Iranian family in which optic atrophy runs as an autosomal dominant trait with an age of onset at 14-15years. We performed linkage analysis with markers mapping to 3q28 and 18q12.2-q12.3 and found linkage to 3q28. Subsequent sequencing of OPA1 identified a novel heterozygous missense mutation (c.1313A>G) replacing aspartic acid by glycine (p.D438G) in the GTPase domain of OPA1. Interestingly, another missense mutation at the same position (c.1313A>T, D438V) has been reported before in two unrelated German families, indicating a possible mutation hot spot. Further evidence supporting the importance of D438 is its conservation from human to acoelomata. OPA1 is believed to be the human orthologue of yeast MGM1, a dynamin-related protein required for the integrity of mitochondrial DNA. Homology modeling of the OPA1 GTPase domain revealed extensive structural similarity to the Dictyostelium dynamin A GTPase domain and showed that D438 may interact with residues of the G1 and the G4 motifs, which are crucial in coordinating GTP. Based on this analysis, we propose a mechanism which explains the gradual decline of vision in ADOA patients with OPA1 mutations at position 438. PMID- 16698015 TI - Captopril suppresses inflammation in endotoxin-induced uveitis in rats. AB - Captopril is an inhibitor of angiotensin-converting enzyme (ACE) that is largely used in the treatment of cardiovascular diseases. Several previous studies have demonstrated that captopril exhibits a wide variety of biological activities, including an anti-inflammatory action, on which we focused our attention. The aim of the present study was to investigate the efficacy of captopril on endotoxin induced uveitis (EIU) in rats. We investigated its effect upon cellular infiltration and protein leakage, as well as on the concentration of tumor necrosis factor-alpha (TNF-alpha), nitric oxide (NO), prostaglandin E2 (PGE2), monocyte chemoattractant protein-1 (MCP-1) in the anterior chamber. In addition, we checked its effect on activation of nuclear factor kappa B (NF-kappaB) in iris and ciliary body (ICB) cells in vivo. EIU was induced in male Lewis rats by a footpad injection of lipopolysaccharide (LPS). One hour after the LPS inoculation, either 1mg/kg, 10mg/kg or 100mg/kg captopril were injected intravenously. 24h later, the aqueous humor was collected from both eyes, and the number of infiltrating cells and protein concentration in the aqueous humor were determined. Levels of TNF-alpha, PGE2, NO and MCP-1 were determined by enzyme linked immunosorbent assay. On some eyes, after enucleation, immunohistochemical staining with a monoclonal antibody against activated NF-kappaB was performed. Captopril treatment significantly decreased the inflammatory cells infiltration, the level of protein, concentrations of TNF-alpha, PGE2, NO and MCP-1 in the aqueous humor. The number of activated NF-kappaB-positive cells was lower in ICB of the rats treated with captopril 3h after the LPS injection. The present results indicate that captopril suppresses the inflammation in EIU by inhibiting the NF-kappaB-dependent pathway and the subsequent production of pro-inflammatory mediators. PMID- 16698016 TI - Different patterns of axonal damage after intracerebral injection of malonate or AMPA. AB - White matter damage occurs following stroke and traumatic brain injury. In preclinical studies of potential therapies to reduce acute brain damage, it is important not only to understand the mechanisms by which this damage occurs, but also to employ techniques that fully quantify the extent of damage. In both respects, neurons have previously received greater attention than axons. The aim of the present study was to compare the extent of axonal damage visualised with different immunohistochemical markers following intracerebral injection of either the excitotoxin AMPA or the mitochondrial inhibitor malonate. Adult mice received intrastriatal injection of toxin and 24 h later the amount of white matter damage visualised with either amyloid precursor protein (APP) or neurofilament 200 (NF200) immunohistochemistry. Malonate induced a dose-dependent increase in the extent of axonal damage with either marker. However, AMPA induced a dose dependent increase in the extent of axonal damage visualised by NF200 immunoreactivity but not by APP immunoreactivity. Malonate and AMPA also differed in their effects on other assessments of white matter integrity and (14)C-2 deoxyglucose autoradiography revealed the two toxins to differ in their initial effects on cerebral metabolism. These data indicate that the ability of commonly used axonal damage markers to quantify the full extent of white matter damage differs following initial excitotoxicity or mitochondrial inhibition. We also confirmed that the markers reveal different extents of axonal damage in a rat model of focal cerebral ischaemia. Therefore, in preclinical studies designed to assess brain protecting agents, it is advisable to use more than one marker to quantify the true extent of axonal damage. PMID- 16698018 TI - Serum from pemphigus vulgaris reduces desmoglein 3 half-life and perturbs its de novo assembly to desmosomal sites in cultured keratinocytes. AB - Defects of cell-cell adhesion underlie disruption of epithelial integrity observed in patients with pemphigus vulgaris (PV), an autoimmune disease characterized by severe mucosal erosions and skin blisters. Pathogenic PV autoantibodies found in patients' sera target desmoglein 3 (Dsg3), a major component of the desmosome, but how does this phenomenon affect Dsg-dependent adhesion and lead to acantholysis still remains controversial. Here, we show that PV serum determines a reduction of Dsg3 half-life in HaCaT keratinocytes, although the total amount of Dsg3 remains unchanged. Immunofluorescence studies suggest that PV IgG exert their effect prevalently by binding non-desmosomal Dsg3 without causing its massive internalization. Furthermore, PV IgG targeting desmosome-assembled Dsg3 do not induce depletion of Dsg3 from the adhesion sites. Conversely, incorporation of PV IgG-Dsg3 complexes into new forming desmosomes appears perturbed. With our study, the basic biochemical changes of Dsg3 in an in vitro model of PV have been defined. PMID- 16698017 TI - Nobiletin, a citrus flavonoid, suppresses phorbol ester-induced expression of multiple scavenger receptor genes in THP-1 human monocytic cells. AB - Unregulated uptake of oxidized low-density lipoproteins (ox-LDL) via macrophage scavenger receptors (SRs) such as lectin-like ox-LDL receptor-1 (LOX-1) is a key event in atherosclerosis. In this study, we examined the effects of five selected food phytochemicals on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced LOX-1 mRNA expression in THP-1 human monocyte-like cells. Nobiletin, a citrus polymethoxylated flavone, markedly reduced it in dose- and time-dependent manners. It also suppressed the phosphorylation of extracellular signal-regulated protein kinase (ERK) 1/2, c-Jun NH2-terminal kinase (JNK) 1/2, and c-Jun (Ser 63), thereby inhibiting the transcriptional activity of activator protein-1. Further nobiletin attenuated expression of SR-A, SR-PSOX, CD36, and CD68, but not CLA-1, mRNA, leading to the blockade of DiI-acLDL uptake. Together, our results suggest that nobiletin is a promising phytochemical for regulating atherosclerosis with reasonable action mechanisms. PMID- 16698019 TI - Fatty acid control of nitric oxide production by macrophages. AB - Modulation of macrophage functions by fatty acids (FA) has been studied by several groups, but the effect of FA on nitric oxide production by macrophages has been poorly examined. In the present study the effect of palmitic, stearic, oleic, linoleic, arachidonic, docosahexaenoic and eicosapentaenoic acids on NF kappaB activity and NO production in J774 cells (a murine macrophage cell line) was investigated. All FA tested stimulated NO production at low doses (1-10 microM) and inhibited it at high doses (50-200 microM). An increase of iNOS expression and activity in J774 cells treated with a low concentration of FA (5 microM) was observed. The activity of NF-kappaB was time-dependently enhanced by the FA treatment. The inhibitory effect of FA on NO production may be due to their cytotoxicity, as observed by loss of membrane integrity and/or increase of DNA fragmentation in cells treated for 48 h with high concentrations. The results indicate that, at low concentrations FA increase NO production by J774 cells, whereas at high concentrations they cause cell death. PMID- 16698020 TI - Interaction of pro-apoptotic protein HGTD-P with heat shock protein 90 is required for induction of mitochondrial apoptotic cascades. AB - HGTD-P is a hypoxia-responsive pro-apoptotic protein that transmits hypoxic signals directly to mitochondria. When overexpressed, HGTD-P induces cell death via typical mitochondrial apoptotic cascades. However, much is unknown about post transcriptional modification and signaling networks of HGTD-P in association with cell death-regulating proteins. We performed yeast two-hybrid screening to identify the molecules involved in HGTD-P-mediated cell death pathways. In this study, we show that heat shock protein 90 physically interacts with HGTD-P and that suppression of Hsp90 activity by low concentrations of geldanamycin reduced HGTD-P-induced mitochondrial catastrophe through inhibition of mitochondrial translocation of HGTD-P. PMID- 16698021 TI - Analysis of leptin signalling in hematopoietic cells using an adapted MAPPIT strategy. AB - The adipocyte-secreted hormone leptin participates in the regulation of hematopoiesis and enhances proliferation of hematopoietic cells. We used an adaptation of the MAPPIT mammalian two-hybrid method to study leptin signalling in a hematopoietic setting. We confirmed the known interactions of suppressor of cytokine signalling 3 (SOCS3) and STAT5 with the Y985 and Y1077 motifs of the leptin receptor, respectively. We also provide evidence for novel interactions at the Y1077 motif, including phospholipase C gamma and several members of the SOCS protein family, further underscoring the important role of the Y1077 motif in leptin signalling. PMID- 16698022 TI - POSH, a scaffold protein for JNK signaling, binds to ALG-2 and ALIX in Drosophila. AB - Plenty of SH3s (POSH) functions as a scaffold protein for the Jun N-terminal kinase (JNK) signal transduction pathway, which leads to cell death in mammalian cultured cells and Drosophila. Here, we show that POSH forms a complex with Apoptosis-linked gene-2 (ALG-2) and ALG-2-interacting protein (ALIX/AIP1) in a calcium-dependent manner. Overexpression of ALG-2 or ALIX in developing imaginal eye discs resulted in roughened or melanized eyes, respectively. These phenotypes were enhanced by co-overexpression of POSH. We found that overexpression of either gene could induce ectopic JNK activation, suggesting that POSH/ALG-2/ALIX may function together in the regulation of the JNK pathway. PMID- 16698023 TI - PFKFB3 gene silencing decreases glycolysis, induces cell-cycle delay and inhibits anchorage-independent growth in HeLa cells. AB - The high rate of glycolysis despite the presence of oxygen in tumor cells (Warburg effect) suggests an important role for this process in cell division. The glycolytic rate is dependent on the cellular concentration of fructose 2,6 bisphosphate (Fru-2,6-P2), which, in turn, is controlled by the bifunctional enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK-2). The ubiquitous PFK-2 isoenzyme (uPFK-2, alternatively named UBI2K5 or ACG) coded by the pfkfb3 gene is induced by different stimuli (serum, progesterone, insulin, hypoxia, etc.) and has the highest kinase/phosphatase activity ratio amongst all PFK-2 isoenzymes discovered to date, which is consistent with its role as a powerful activator of glycolysis. uPFK-2 is expressed in brain, placenta, transformed cells and proliferating cells. In the present work, we analyze the impact of small interfering RNA (siRNA)-induced silencing of uPFK-2 on the inhibition of cell proliferation. HeLa cells treated with uPFK-2 siRNA showed a decrease in uPFK-2 RNA levels measured at 24h. uPFK-2 protein levels were severely depleted at 48-72h when compared with cells treated with an unrelated siRNA, correlating with decreased glycolytic activity, Fru-2,6-P2, lactate and ATP concentrations. These metabolic changes led to reduced viability, cell-cycle delay and an increase in the population of apoptotic cells. Moreover, uPFK-2 suppression inhibited anchorage-independent growth. The results obtained highlight the importance of uPFK-2 on the regulation of glycolysis, on cell viability and proliferation and also on anchorage-independent growth. These data underscore the potential for uPFK-2 as an effective tumor therapeutic target. PMID- 16698024 TI - Application of the transtheoretical model to identify psychological constructs influencing exercise behavior: a questionnaire survey. AB - BACKGROUND: Current research on exercise behavior has largely been attempted to identify the relationship between psychological attributes and the initiation or adherence of exercise behavior based on psychological theories. A limited data are available on the psychological predictors of exercise behavior in public health. OBJECTIVES: The present study examined the theorized association of TTM of behavior change constructs by stage of change for exercise behavior. METHODS: A total of 228 college students selected from 2 universities in Seoul were surveyed. Four Korean-version questionnaires were used to identify the stage of exercise behavior and psychological attributes of adolescents. Data were analyzed by frequency analysis, MANOVA, correlation analysis, and discriminant function analysis. RESULTS: Multivariate F-test indicated that behavioral and cognitive processes of change, exercise efficacy, and pros differentiated participants across the stages of exercise behavior. Furthermore, exercise behavior was significantly correlated with the TTM constructs, and that overall classification accuracy across the stages of change was 61.0%. CONCLUSIONS: The present study supports the internal and external validity of the Transtheoretical Model for explaining exercise behavior. As this study highlights, dissemination must increase awareness but also influences perceptions regarding theoretically based and practically important exercise strategies for public health professionals. PMID- 16698025 TI - Genotyping Trichomonas vaginalis. AB - A genotyping method has been developed to distinguish each Trichomonas vaginalis isolate and has provided the first genome mapping studies of this protist with an estimated 180Mb genome. The technique was developed using high molecular weight DNA prepared from five laboratory isolates from Australia and USA and 20 clinical isolates from South Africa. Inhibition of the notorious T. vaginalis endogenous nucleases by addition of potent inhibitors was essential to the success of this study. Chromosomal DNA larger than 2.2Mb was macrorestricted to a minimum segment size of approximately 50kb, separated by pulsed field gel electrophoresis and hybridised with a variety of gene probes. Each isolate generated a unique pattern that was distinguished by each of the probes. Four single copy gene probes (fd, hmp35, ibp39 and pfoD) were identified but probes which identified several bands (pfoB and alpha-scs) per isolate were most informative for genotyping. The pyruvate:ferredoxin oxidoreductase B gene probe identified two to seven copies of pfoB (or its closely related homologue pfoA) per genome in different isolates and is an obvious candidate probe to identify epidemiological linkage between infections by this genotyping method. Cleavage of the genomes into smaller fragments failed to distinguish isolates from diverse locations indicating the proximal regions of genes are conserved. PMID- 16698026 TI - Measurement of intracellular strain on deformable substrates with texture correlation. AB - Mechanical stimuli are important factors that regulate cell proliferation, survival, metabolism and motility in a variety of cell types. The relationship between mechanical deformation of the extracellular matrix and intracellular deformation of cellular sub-regions and organelles has not been fully elucidated, but may provide new insight into the mechanisms involved in transducing mechanical stimuli to biological responses. In this study, a novel fluorescence microscopy and image analysis method was applied to examine the hypothesis that mechanical strains are fully transferred from a planar, deformable substrate to cytoplasmic and intranuclear regions within attached cells. Intracellular strains were measured in cells derived from the anulus fibrosus of the intervertebral disc when attached to an elastic silicone membrane that was subjected to tensile stretch. Measurements indicated cytoplasmic strains were similar to those of the underlying substrate, with a strain transfer ratio (STR) of 0.79. In contrast, nuclear strains were much smaller than those of the substrate, with an STR of 0.17. These findings are consistent with previous studies indicating nuclear stiffness is significantly greater than cytoplasmic stiffness, as measured using other methods. This study provides a novel method for the study of cellular mechanics, including a new technique for measuring intranuclear deformations, with evidence of differential magnitudes and patterns of strain transferred from the substrate to cell cytoplasm and nucleus. PMID- 16698027 TI - New approach to calculating and predicting the ionic strength generated during carrier ampholyte isoelectric focusing. AB - A serious drawback of the carrier ampholyte isoelectric focusing is the undetermined ionic strength at which the proteins separate. We tried to study its effect in order to bridge the gap between theory and practice giving a quantitative and qualitative description. Using certain electrode systems, we managed to calculate the discrete values of the ionic strength in a random position between the electrodes. As a rule, higher values of up to 10(-3)mol/l were obtained near the electrodes, which gradually decreased towards the middle of the carrier reaching 10(-7)mol/l. A relationship between the measured isoelectric points (pI) of some proteins and the ionic strength was found. In the present paper, we report our findings on the dynamics of the process referring to the origin of ionic strength, its formation and alteration during the isoelectric focusing process. PMID- 16698028 TI - Development of an ion chromatographic gradient retention model from isocratic elution experiments. AB - When facing separation problems in ion chromatography, chromatographers often lack guidelines to decide a priori if isocratic elution will give enough separation in a reasonable analysis time or a gradient elution will be required. This situation may be solved by the prediction of retention in gradient elution mode by using isocratic experimental data. This work describes the development of an ion chromatographic gradient elution retention model for fluoride, chloride, nitrite, bromide, nitrate, sulfate and phosphate by using isocratic experimental data. The isocratic elution retention model was developed by applying a polynomial relation between the logarithm of the retention factor and logarithm of the concentration of competing ions; the gradient elution retention model was based on the stepwise numerical integration of the corresponding differential equation. It was shown that the developed gradient elution retention model was not significantly affected by transferring data form isocratic experiment. The root mean squared prediction error for gradient elution retention model was between 0.0863 for fluoride and 0.7027 for bromide proving a very good predictive ability of developed gradient elution retention model. PMID- 16698029 TI - An assessment of the retention behaviour of polycyclic aromatic hydrocarbons on reversed phase stationary phases: selectivity and retention on C18 and phenyl type surfaces. AB - In this manuscript the retention and selectivity of a set of linear and non linear PAHs were evaluated on five different reversed-phase columns. These phases included C18 and C18 Aqua stationary phases, as well as three phenyl phases: Propyl-phenyl, Synergi polar-RP and Cosmosil 5PBB phase. Overall, the results revealed that the phenyl-type columns offered better separation performance for the linear PAHs, while the separation of the structural isomer PAHs was enhanced on the C18 columns. The Propyl-phenyl column was found to have the highest molecular-stationary phase interactions, as evidenced by the greatest rate of change in 'S' (0.71) as a function of the molecular weight in the PAH homologous series, despite having the lowest surface coverage (3% carbon load) (where S is the slope of a plot of logk versus the solvent composition). In contrast, the C18 Aqua column, having the highest surface coverage (15% carbon load) was found to have the second lowest molecular-stationary phase interactions (rate of change in S=0.61). Interestingly, the Synergi polar-RP column, which also is a phenyl stationary phase behaved more 'C18-like' than 'phenyl-like' in many of the tests undertaken. This is probably not unexpected since all five phases were reversed phase. PMID- 16698030 TI - Surface modification of phenolphthalein poly(ether sulfone) ultrafiltration membranes by blending with acrylonitrile-based copolymer containing ionic groups for imparting surface electrical properties. AB - Asymmetric ultrafiltration membranes were fabricated from the blends of phenolphthalein polyethersulfone (PES-C) and acrylonitrile copolymers containing charged groups, poly(acrylonitrile-co-acrylamido methylpropane sulfonic acid) (PAN-co-AMPS). From the surface analysis by XPS and ATR-FTIR, it was found that the charged groups tend to accumulate onto the membrane surface. This result indicated that membrane surface modification for imparting surface electrical properties could be carried out by blending charged polymer. Furthermore, with the help of a relatively novel method to measure membrane conduction, the true zeta potentials calculated on the basis of the streaming potential measurements were used to reflect the charge state of membrane surface. In addition, it was noteworthy that, from the profiles of zeta potential versus pH curves and the magnitude of zeta potentials, the determination of zeta potential was dependent not only on the electrical properties of membrane surface but also on its hydrophilicity. At last, based on a relatively elaborate study on the electrostatic interaction between the membrane surface and protein, it was found that these charged membranes could meet different demands of membrane applications, such as resisting protein fouling or protein separation, through adjusting solution pH value. PMID- 16698031 TI - Molecular characterization of hemoglobin from the honeybee Apis mellifera. AB - Due to the prevailing importance of the tracheal system for insect respiration, hemoglobins had been considered rare exceptions in this arthropod subphylum. Here we report the identification, cloning and expression analysis of a true hemoglobin gene in the honeybee Apis mellifera (Hymenoptera). The deduced amino acid sequence covers 171 residues (19.5kDa) and harbors all globin-typical features, including the proximal and the distal histidines. The protein has no signal peptide for transmembrane transport and was predicted to localize in the cytoplasm. The honeybee hemoglobin gene shows an ancient structure, with introns in positions B12.2 and G7.0, while most other insect globins have divergent intron positions. In situ hybridization studies showed that hemoglobin expression in the honeybee is mainly associated with the tracheal system. We also observe hemoglobin expression in the Malpighi tubes and testis. We further demonstrated that hemoglobins occur in other insect orders (Hemiptera, Coleoptera, Lepidoptera), suggesting that such genes belong to the standard repertoire of an insect genome. Phylogenetic analyses show that globins evolved along with the accepted insect systematics, with a remarkable diversification within the Diptera. Although insect hemoglobins may be in fact involved in oxygen metabolism, it remains uncertain whether they carry out a myoglobin-like function in oxygen storage and delivery. PMID- 16698032 TI - A CCR1 antagonist prevents the development of experimental autoimmune myocarditis in association with T cell inactivation. AB - Chemokines play an important role in induction of chemotaxis of immune cells. CCR1 is a chemokine receptor expressed on neutrophils, monocytes, and T lymphocytes. The role of CCR1 in immunity is not well examined. We demonstrated the role of CCR1 on T lymphocytes and the effect of a CCR1 antagonist, BX471 in myocarditis. Lewis rats were immunized with cardiac myosin on day 0 to establish experimental autoimmune myocarditis. Rats were then administered BX471 subcutaneously every day (group BX0: n = 7) or from day 14 (group BX14: n = 7) and were killed on day 21. We confirmed expression of CCR1 in cells infiltrating the myocardium by immunohistochemistry and FACS analysis. The development of myocarditis was almost completely prevented in group BX0, and myocarditis affected areas were significantly decreased in size in group BX14. Cardiac function was markedly improved. Ribonuclease protection assay showed that the CCR1 antagonist treatment suppressed mRNA expression for IL-6, IL-1beta, and TNF alpha in the hearts. An antigen-specific T cell proliferation assay was performed with CD4-positive T cells isolated from control rats immunized with cardiac myosin. T cell proliferation was inhibited by the CCR1 antagonist. Additionally, we showed by Western blot that the CCR1 antagonist suppressed ERK1/2 and JNK activities in T cells stimulated with myosin and that IL-2 reversed this suppression. The CCR1 antagonist reduced the severity of EAM by inhibiting cytokine expression and inducing T cell inactivation. Thus, the CCR1 antagonist may provide a novel therapeutic strategy treatment of myocarditis. PMID- 16698033 TI - PPARdelta modulates lipopolysaccharide-induced TNFalpha inflammation signaling in cultured cardiomyocytes. AB - Peroxisome proliferator activated receptors (PPARs: PPARalpha, gamma and delta) regulate fatty acid metabolism, glucose homeostasis, cell proliferation, differentiation and inflammation. Tumor necrosis factor alpha (TNFalpha) is one of the important pathological factors in inflammatory responses during the pathological progression of myocardial ischemic/reperfusion and hypertrophy. Accumulating evidence shows that synthetic ligands of PPARalpha and PPARgamma suppress myocardial inflammatory responses, such as the production of TNFalpha, thus exerting beneficial effects in animals who had undergone ischemia/reperfusion injury or cardiac hypertrophy. However, it remains obscured if PPARdelta and its ligands exert any effect on the production of TNFalpha, thus influencing cardiac inflammatory responses. In this study, we investigated the effects of PPARdelta and its synthetic ligand GW0742 on TNFalpha production in cultured cardiomyocytes. Our studies indicate that a PPARdelta-selective ligand inhibited lipopolysaccharide (LPS)-induced TNFalpha production from cardiomyocytes. Adenoviral-mediated overexpression of PPARdelta substantially inhibited TNFalpha expression in cultured cardiomyocytes compared to controls, whereas overexpression of a PPARdelta mutant with ablated ligand binding domain did not show similar effect. Conversely, absence of PPARdelta in cardiomyocytes further exaggerated LPS-induced TNFalpha production. Moreover, activation of PPARdelta abrogated LPS-induced degradation of IkappaBs, thus suppressing LPS induced nuclear factor kappaB (NF-kappaB) activities. Therefore, PPARdelta is an important determinant of TNFalpha expression via the NF-kappaB signaling pathway, thus serving as therapeutic targets to attenuate inflammation that are involved in cardiac pathological progression. PMID- 16698034 TI - Hijacking of a substrate-binding protein scaffold for use in mycobacterial cell wall biosynthesis. AB - The waxy cell wall is crucial to the survival of mycobacteria within the infected host. The cell wall is a complex structure rich in unusual molecules that includes two related lipoglycans, the phosphatidylinositol mannosides (PIMs) and lipoarabinomannans (LAMs). Many proteins implicated in the PIM/LAM biosynthetic pathway, while attractive therapeutic targets, are poorly defined. The 2.4A resolution crystal structure of an essential lipoprotein, LpqW, implicated in LAM biosynthesis is reported here. LpqW adopts a scaffold reminiscent of the distantly related, promiscuous substrate-binding proteins of the ATP-binding cassette import system. Nevertheless, the unique closed conformation of LpqW suggests that mycobacteria and other closely related pathogens have hijacked this scaffold for use in key processes of cell wall biosynthesis. In silico docking provided a plausible model in which the candidate PIM ligand binds within a marked electronegative region located on the surface of LpqW. We suggest that LpqW represents an archetypal lipoprotein that channels intermediates from a pathway for mature PIM production into a pathway for LAM biosynthesis, thus controlling the relative abundance of these two important components of the cell wall. PMID- 16698035 TI - ACCA phosphopeptide recognition by the BRCT repeats of BRCA1. AB - The tumour suppressor gene BRCA1 encodes a 220 kDa protein that participates in multiple cellular processes. The BRCA1 protein contains a tandem of two BRCT repeats at its carboxy-terminal region. The majority of disease-associated BRCA1 mutations affect this region and provide to the BRCT repeats a central role in the BRCA1 tumour suppressor function. The BRCT repeats have been shown to mediate phospho-dependant protein-protein interactions. They recognize phosphorylated peptides using a recognition groove that spans both BRCT repeats. We previously identified an interaction between the tandem of BRCA1 BRCT repeats and ACCA, which was disrupted by germ line BRCA1 mutations that affect the BRCT repeats. We recently showed that BRCA1 modulates ACCA activity through its phospho-dependent binding to ACCA. To delineate the region of ACCA that is crucial for the regulation of its activity by BRCA1, we searched for potential phosphorylation sites in the ACCA sequence that might be recognized by the BRCA1 BRCT repeats. Using sequence analysis and structure modelling, we proposed the Ser1263 residue as the most favourable candidate among six residues, for recognition by the BRCA1 BRCT repeats. Using experimental approaches, such as GST pull-down assay with Bosc cells, we clearly showed that phosphorylation of only Ser1263 was essential for the interaction of ACCA with the BRCT repeats. We finally demonstrated by immunoprecipitation of ACCA in cells, that the whole BRCA1 protein interacts with ACCA when phosphorylated on Ser1263. PMID- 16698037 TI - Obstetrical complications in children at high risk for bipolar disorder. AB - OBJECTIVE: To examine obstetrical complications as a risk factor for developing bipolar disorder (BPD). We hypothesized that children with a bipolar parent would be at greater risk for obstetrical complications than demographically matched children of healthy adults. Additionally, within this "at-risk" (AR) sample, we hypothesized that obstetrical complications would be associated with the development of psychiatric disorders. METHODS: The Washington University in St. Louis Kiddie-Schedule for Affective Disorders and Schizophrenia (WASH-U KSADS) was administered to children (AR) who had at least one parent with BPD (N=36) and children of healthy parents (HC) (N=27), by raters who were blind to diagnostic category. To assess obstetrical risk history, the Rochester Research Obstetrical Scale (ROS) was administered to parents of AR and HC children. RESULTS: Children at familial risk for BPD had greater total (p=0.02) and prenatal (p=0.006) obstetrical complication scores than children of healthy parents. However, obstetrical complications were not associated with the development of affective, anxiety, or disruptive behavioral disorders within the at-risk group. CONCLUSION: Our data suggest that compared with children of families without BPD, children of parents with BPD may be at greater risk for obstetrical complications, particularly those that occur during the prenatal period; however, at this early follow-up period factors other than obstetrical complications appear to contribute to the differences in rates of psychiatric disorders between these groups. PMID- 16698036 TI - Crystallographic structure of human beta-hexosaminidase A: interpretation of Tay Sachs mutations and loss of GM2 ganglioside hydrolysis. AB - Lysosomal beta-hexosaminidase A (Hex A) is essential for the degradation of GM2 gangliosides in the central and peripheral nervous system. Accumulation of GM2 leads to severely debilitating neurodegeneration associated with Tay-Sachs disease (TSD), Sandoff disease (SD) and AB variant. Here, we present the X-ray crystallographic structure of Hex A to 2.8 A resolution and the structure of Hex A in complex with NAG-thiazoline, (NGT) to 3.25 A resolution. NGT, a mechanism based inhibitor, has been shown to act as a chemical chaperone that, to some extent, prevents misfolding of a Hex A mutant associated with adult onset Tay Sachs disease and, as a result, increases the residual activity of Hex A to a level above the critical threshold for disease. The crystal structure of Hex A reveals an alphabeta heterodimer, with each subunit having a functional active site. Only the alpha-subunit active site can hydrolyze GM2 gangliosides due to a flexible loop structure that is removed post-translationally from beta, and to the presence of alphaAsn423 and alphaArg424. The loop structure is involved in binding the GM2 activator protein, while alphaArg424 is critical for binding the carboxylate group of the N-acetyl-neuraminic acid residue of GM2. The beta subunit lacks these key residues and has betaAsp452 and betaLeu453 in their place; the beta-subunit therefore cleaves only neutral substrates efficiently. Mutations in the alpha-subunit, associated with TSD, and those in the beta subunit, associated with SD are discussed. The effect of NGT binding in the active site of a mutant Hex A and its effect on protein function is discussed. PMID- 16698038 TI - Assessment of cerebral blood volume in schizophrenia: A magnetic resonance imaging study. AB - Brain atrophy has consistently been observed in schizophrenia, representing a 'gross' evidence of anatomical abnormalities. Reduced cerebral blood volume (CBV) may accompany brain size decrement in schizophrenia, as suggested by prior small SPECT studies. In this study, we non-invasively investigated the hemisphere CBV in a large sample of patients suffering from schizophrenia with perfusion weighted imaging (PWI). PWI images were obtained, following intravenous injection of paramagnetic contrast agent (Gadolinium-DTPA), for 54 DSM-IV patients with schizophrenia (mean age+/-SD=39.19+/-12.20 years; 34 males, 20 females) and 24 normal controls (mean age+/-SD=44.63+/-10.43 years; 9 males, 15 females) with a 1.5T Siemens magnet using an echo-planar sequence (TR=2160 ms, TE=47 ms, slice thickness=5mm). The contrast of enhancement (CE), a semi-quantitative parameter inversely estimating the CBV, were calculated pixel by pixel as the ratio of the maximum signal intensity drop during the passage of contrast agent (Sm) by the baseline pre-bolus signal intensity (So) (CE=Sm/Sox100) for right and left hemisphere on two axial images. Specifically, higher CE values correspond to lower CBV and viceversa Compared to normal controls, patients with schizophrenia had significantly higher bilateral hemisphere CE values (p=0.02) and inverse CE laterality index (p=0.02). This study showed abnormally reduced and inverse hemisphere CBV in a large population of patients with schizophrenia. Hypothetically, chronic low CBV may sustain neural hypoactivation and concomitant increase of free radicals, ultimately resulting in neuronal loss and cognitive impairments. Thus, altered intracranial hemodynamics may accompany brain atrophy and cognitive deficits, being a crucial factor in the pathophysiology of schizophrenia. PMID- 16698039 TI - Designing sensor systems capable of differentiating children from adults. AB - INTRODUCTION: Injury prevention systems intended to prevent children from entering hazardous locations (or at least alert caregivers if that occurs) often respond to every instance of a person's presence, regardless of whether the intruder is a child. This performance results in a high nuisance alarm rate that sometimes causes adults to disable or circumvent the safety system. If a child safety system can accurately identify intruders as adults or children, nuisance alarm rates can be decreased. METHOD: This analysis selects three human factors (height, foot length, and cognition) amenable to adult/child differentiation and describes likely sensor strategies, advantages, and disadvantages. RESULTS: Preliminary testing of prototypes systems shows that simple sensor systems are capable of acquiring adequate data for adult/child differentiation. The discussion addresses requirements for discriminator systems and the effects of various sensor combinations on overall performance. PMID- 16698040 TI - Construction of genetic oscillators with interlocked feedback networks. AB - To understand how a gene regulatory network functioning as an oscillator is built, a precise mathematical description of the network and its dynamical properties are developed in this paper. Our approach is based on analyzing the effect of interactions between smaller subnetworks with simple dynamics. We relate an oscillatory behavior of the network to the destabilization phenomenon of a steady state caused by the interactions in a simple discrete map. When source of the instability of the steady state rather than the oscillatory behavior itself is considered, linear stability analysis and feedback control theory can be employed. Moreover, the amplitudes robust against change in delay can also be obtained from the discrete map. The main ideas are illustrated by constructing a genetic oscillator termed a "repressilator" and analyzing the occurrence of cellular rhythms, although the theoretical results hold for a general class of biological systems. The method can be directly applied to design, construct genetic oscillators, and further control their dynamics, even for large-scale networks. PMID- 16698041 TI - Maintenance of brain thyroid hormone level during peripheral hypothyroid condition in adult rat. AB - Thyroid hormones are essential for normal functioning of adult mammalian brain. The present investigation deals with the understanding of the time course of thyroid hormone homeostasis in adult rat brain. Animals were rendered hypothyroid by PTU injections (2 mg/100 g bw) for 30 consecutive days. Serum and synaptosomal T3/T4 content, synaptosomal AChE and Na+-K+-ATPase activities were determined on alternate days. While serum T4 level initially increased on the second day compared to control, serum T3 declined in a triphasic pattern; the first phase lasting from the second day to the 6th day, the second phase ended on the 14th day and last phase continued till the 30th day. Cerebro-cortical synaptosomal T3 level increased on the 2nd day from the control, attained a peak on the 4th day, remained stable until the 18th day, and abruptly declined on the 20th day. Synaptosomal T4 content remained negligible or undetected throughout. Synaptosomal membrane Na+-K+-ATPase and AChE activity exhibited an inverse relationship during the experimental regime, being much more prominent on the 2nd, 18th and 20th day coinciding with the variations in brain T3 level. Thus, the study identifies the onset of central homeostasis between the first and second day, its continuation for about 16-18 days and its termination between the 18th and 20th day. PMID- 16698042 TI - Effect of chronic supplementation with branched-chain amino acids on the performance and hepatic and muscle glycogen content in trained rats. AB - The objective of this study was to evaluate the effects of a diet supplemented with branched-chain amino acids (BCAA; 3.57% and 4.76%) on the performance and glycogen metabolism of trained rats. Thirty-six adult male Wistar rats received the control diet (AIN-93M) (n=12) and two diets supplemented with BCAA (S1: AIN 93M+3.57% BCAA, n=12, and S2: AIN-93M+4.76% BCAA, n=12) for 6 weeks. The training protocol consisted of bouts of swimming exercise (60 min day(-1)) for 6 weeks at intensities close to the lactate threshold. On the last day of the experiment, all groups were trained for 1 h (1H) or were submitted to the exhaustion test (EX). The time to exhaustion did not differ between groups. The groups submitted to the exhaustion test presented a reduction in plasma glucose and an increase in plasma ammonia and blood lactate concentrations compared to the 1H condition. In the 1H condition, hepatic glycogen concentration was significantly higher in group S2 compared to the control diet and S1 groups (132% and 44%, respectively). Group S2 in the 1H condition presented a higher muscle glycogen concentration (45%) compared to the control diet group. In the EX condition, a significantly higher hepatic glycogen concentration was observed for group S2 compared to the control diet and S1 groups (262% and 222%, respectively). Chronic supplementation with BCAA promoted a higher hepatic and muscle glycogen concentration in trained animals, with this effect being dose dependent. PMID- 16698043 TI - Melatonin directly scavenges free radicals generated in red blood cells and a cell-free system: chemiluminescence measurements and theoretical calculations. AB - Melatonin, a pineal secretory product, has properties of both direct and indirect powerful antioxidant. The aim of the present study was to compare the radical scavenging, structural and electronic properties of melatonin and tryptophan, precursor of melatonin. Using the alkoxyl- and peroxyl radical-generating systems [the organic peroxide-treated human erythrocytes and a cell-free system containing the azo-initiator 2,2'-azobis(2-amidinopropane)dihydrochloride], we evaluated the radical-scavenging effects of melatonin and tryptophan. Melatonin rather than tryptophan at concentrations of 100-2000 microM markedly inhibited membrane lipid peroxidation in human erythrocytes treated with organic hydroperoxide as well as radical-induced generation of luminol-dependent chemiluminescence. The apparent Stern-Volmer constants for inhibition of membrane lipid peroxidation by melatonin and tryptophan were estimated to be (0.23+/-0.05) x 10(4) M(-1) and (0.02+/-0.005) x 10(4) M(-1), respectively. The apparent Stern Volmer constants for inhibition of azo-initiator-derived peroxyl radical generation by melatonin and tryptophan were determined to be (0.42+/-0.05) x 10(4) M(-1) and (0.04+/-0.01) x 10(4) M(-1), respectively. The structural and electronic properties of melatonin and its precursor, tryptophan, were determined theoretically by performing semi-empirical and ab initio calculations. The high radical-scavenging properties of melatonin may be explained by the high surface area value and high dipole moment value. From the thermodynamic standpoint, based on our calculations, N(1)-acetyl-N(2)-formyl-5-methoxykynuramine (AFMK), was the most stable end oxidative product of melatonin. PMID- 16698044 TI - Evaluation of possible mechanisms of protective role of Tamarix gallica against DEN initiated and 2-AAF promoted hepatocarcinogenesis in male Wistar rats. AB - We have previously reported that Tamarix gallica caused a marked inhibition of thioacetamide-induced hepatotoxicity, oxidative damage and early tumor promotion related events in the liver. These results strongly indicates that T. gallica may have chemopreventive potential. Therefore, in the present study, we examined the inhibitory effects of T. gallica methanolic extract on diethylnitrosamine (DEN) initiated and 2-acetyl aminofluorene (2-AAF) promoted liver carcinogenesis in male Wistar rats. Interestingly, it was found that T. gallica (25 and 50 mg/kg body wt.) resulted in a marked reduction of the incidence of liver tumors. The study was further histologically confirmed. Furthermore to understand the underlying mechanisms of chemopreventive action by T. gallica we evaluated the levels activities of hepatic antioxidant defense enzymes, ornithine decarboxylase activity and hepatic DNA synthesis as a marker for tumor promotion since direct correlation between these marker parameters and carcinogenicity have been well documented. Treatment of male Wistar rats for five consecutive days with 2-AAF i.p. induced significant hepatic toxicity, oxidative stress and hyperproliferation. Pretreatment of T. gallica extract (25 and 50 mg/kg body wt.) prevented oxidative stress by restoring the levels of antioxidant enzymes and also prevented toxicity at both the doses. The promotion parameters induced (ornithine decarboxylase activity and DNA synthesis) by 2-AAF administration in diet with partial hepatectomy (PH) were also significantly suppressed dose dependently by T. gallica. Therefore, we can conclude that ultimately the protection against liver carcinogenesis by T. gallica methanolic extract might be mediated by multiple actions, which include restoration of cellular antioxidant enzymes, detoxifying enzymes, ODC activity and DNA synthesis. PMID- 16698045 TI - Oregon Beach Monitoring Program: Bacterial exceedances in marine and freshwater creeks/outfall samples, October 2002-April 2005. AB - A total of 3086 samples, both marine (2916) and freshwater creeks/outfall samples (170) were collected and analyzed for enterococci during October 2002-April 2005, from 52 designated beaches in Oregon. A total of 3.2% (99/3086) of the samples at 21 beaches exceeded 158 most probable number (MPN)/100 milliliters (mL). The average enterococci levels of these 99 exceedances was 559 MPN/100 mL, with a maximum of 4352 MPN/100 mL (Otter Rock and Ona) and a minimum of 160 MPN/100 mL (Sunset Bay State Park, Bastendorff, and Mill). For marine water, 77/2916 (2.6%) exceeded 158 MPN/100 mL. For freshwater, 22/170 (12.9%) exceeded 158 MPN/100 mL, with a maximum of 587 MPN/100 mL at Sunset Bay. Sixty percent of the marine and 9% of freshwater exceedances occurred during the winter. Seventy-two percent (55/77) of the marine exceedances occurred after rainfall events (0.01-60.0 mm). At Harris and Mill Beaches, cumulative rainfall was highly correlated with bacterial densities, R = 0.7. Rainfall, at both beaches, explained about one-half the variation in log10 bacteria density (R2 = 0.5). Additional monitoring is warranted to further characterize bacterial contamination in Oregon waters. PMID- 16698046 TI - Oceanographic conditions in North and Northwest Iberia and their influence on the Prestige oil spill. AB - Oceanographic conditions at the time of the Prestige oil spill (November 2002) and following months are analyzed based on a set of hydrographic cruises. The ship sank off one of the flanks of the Galician Bank, an offshore seamount, and a major oil spill drifted to the N and NW Iberian coast mainly driven by dominant winds. Coastal circulation was characterized by freshwater plumes and the poleward slope current, and could have affected the fate of the oil spill and influenced stranding places. Seasonal evolution of oceanographic conditions in this particular year is compared with the long-term average and reveals specific features that need to be taken into account in studies of the impact of the oil spill on populations. Spring conditions commenced earlier than other years in the Southern Bay of Biscay, contrastingly in western Iberia. The lack of subsurface intrusion of subtropical waters suggests a low intense penetration of the poleward current in Spanish Biscay slopes. In western Iberia, the slope poleward current observed in late autumn weakens and is exported off slope during upwelling pulses in the spring, with no strong intrusion of the poleward current on the slope at the time of the spring bloom. A description of current velocities near the wreck on the Galician Bank is obtained after the analysis of a mooring line. PMID- 16698047 TI - Monitoring biomarkers in fish (Lepidorhombus boscii and Callionymus lyra) from the northern Iberian shelf after the Prestige oil spill. AB - Hepatic biomarker responses were measured in two demersal fish species (Lepidorhombus boscii and Callionymus lyra) from the northern Iberian shelf associated with the massive Prestige oil spill (POS), five months after the accident. The biomarkers selected were 7-ethoxyresorufin-O-deethylase (EROD), glutathione-S-transferase (GST), glutathione reductase (GR), catalase (CAT), and DNA integrity. Interspecies differences and spatial variations in biomarker responses were observed along the shelf. GST, GR and CAT activities were significantly elevated in L. boscii in the most oil impacted area (Finisterre) and positively correlated (p<0.05) with POS tar aggregate densities. The lack of previous data from the area together with the existence of chronic background pollution of the shelf implies that the observed biomarker responses cannot be solely attributed to the petroleum hydrocarbon components of the spilled oil. This first biological effect assessment showed that L. boscii is a potentially suitable target species to be used in future biomonitoring programmes along the northern Iberian shelf. PMID- 16698048 TI - Molecular genetic alterations of FHIT and p53 genes in benign and malignant thyroid gland lesions. AB - Several oncogenes and tumor-suppressor genes are involved either as early or late event in thyroid gland carcinogenesis. Human FHIT (fragile histidine triad) gene is highly conserved gene whose loss of function may be important in the development and/or progression of various types of cancer. We undertook this study to analyze FHIT and p53 gene status in different benignant and malignant thyroid tumors. Status of these genes as well as intensity of apoptosis was analyzed in tumor tissues by molecular genetic methods, immunohistochemistry, and FACS-scan analysis. The majority of the malignant thyroid cancers displayed aberrant expression of FHIT gene, concominant with p53 gene inactivation. This is followed by low rate of apoptosis, which may be important in the development and/or progression of thyroid cancer. We found higher incidence of p53 mutation and aberrant processing of FHIT mRNA in malignant tumors (papillary, follicular, medullary and anaplastic carcinomas) and in those tumors with distant metastasis. The growth of p53(-)/FHIT(-) follicular carcinoma of human origin was much faster in nude mice than p53(+)/FHIT(+) follicular carcinoma, and mice had shorter survival rate. Our results show a correlation between aberrant FHIT and p53 expression, low rate of apoptosis, and malignancy. Concomitant aberration of FHIT gene and p53 could be responsible for development of highly malignant types of thyroid cancer and may be considered as a prognostic marker for these tumors. PMID- 16698049 TI - Adolescent and adult responsiveness to the incentive value of cocaine reward in mice: role of neuronal nitric oxide synthase (nNOS) gene. AB - A major concern in adolescent psychostimulant abuse is the long-term consequence of this practice, because early drug exposure may cause long-term adaptations, which render the organism more susceptible to drug abuse later in life. The incentive value of drug and natural reward in rodents is commonly assessed by the conditioned place preference (CPP) paradigm, which involves Pavlovian learning. The aims of the present study were to investigate: a) the acquisition, expression, maintenance and reinstatement of cocaine CPP from periadolescence (PD24-45) through adulthood (PD70); b) potential sexual dimorphism in adolescence and adulthood in response to cocaine-induced CPP; and c) the role of the neuronal nitric oxide synthase (nNOS) gene in long-term neural plasticity underlying responsiveness to cocaine and cocaine-associated cues. Adolescent wild type (WT) mice acquired significant cocaine (20 mg/kg) CPP that was maintained from PD24 through PD43. Upon extinction, CPP was reinstated in adulthood (PD70) following a priming injection of cocaine (5 mg/kg). In contrast, cocaine CPP acquired between PD26 and PD31 in adolescent nNOS knockout (KO) mice, was neither maintained nor reinstated by cocaine. There was no sexual dimorphism in adolescent WT and KO mice. Genotype differences and sexual dimorphism were observed in adult mice. Cocaine CPP in adult WT males (PD89-94) was maintained for 4 weeks post training, and subsequently reinstated by cocaine priming; the magnitude of CPP in adult WT males was lower than in female counterparts. CPP in adult KO males (PD88-93) was neither maintained nor reinstated by cocaine priming; in contrast, CPP in adult KO females was not significantly different from adult WT females. Results suggest that the nNOS gene is essential during adolescence of both sexes for the development of long-term neural plasticity underlying responsiveness to the incentive value of cocaine reward. Sexual dimorphism in response to cocaine CPP emerges in adulthood; nNOS contribution to long-term plasticity is therefore sexually dimorphic and age-dependent in female but not in male subjects. PMID- 16698050 TI - Increased prepulse inhibition of the acoustic startle response is associated with better strategy formation and execution times in healthy males. AB - Prepulse inhibition (PPI) refers to the attenuation of the amplitude of the startle reflex in response to sudden intense stimuli (pulse) if preceded by a weaker sensory stimulus (prepulse). PPI reflects the ability to filter out irrelevant information in the early stages of processing so that attention can be directed to more salient environmental features. Inhibition at this early stage of information processing appears modulated by the prefrontal cortex in a "top down" fashion and this may account for the normal inter-individual variability in PPI and in cognitive performance. PPI data were calculated from 82 healthy male subjects who were also tested in problem solving (Stockings of Cambridge; SoC), spatial working memory (SWM) and 5-choice reaction time (RT) tests from the Cambridge Neuropsychological Test Automated Battery. Correlations between PPI scores and cognitive test variables were examined. In addition PPI scores were divided in quartiles which were used as grouping factors in examining cognitive test performance. Compared to individuals in the lowest quartile those in the highest had (a) shorter execution but not reaction times on the 5-choice RT, (b) shorter subsequent but not initial thinking times in the SoC where they also solved more problems correctly with the minimum number of moves, and (c) better strategy but not errors scores in the SWM. Our findings suggest that greater PPI is associated with superior abilities in strategy formation and execution times. We suggest that this is due to more efficient early information processing. PMID- 16698051 TI - Imitation during phoneme production. AB - Does listening to and observing the speaking interlocutor influence phoneme production? In two experiments female participants were required to recognize and, then, to repeat the string-of-phonemes /aba/ presented by actors visually, acoustically and audiovisually. In experiment 1 a male actor presented the string of-phonemes and the participants' lip kinematics and voice spectra were compared with those of a reading control condition. In experiment 2 female and male actors presented the string-of-phonemes and the lip kinematics and the voice spectra of the participants' responses to the male actors were compared with those to the female actors (control condition). In both experiments 1 and 2, the lip kinematics in the visual presentations and the voice spectra in the acoustical presentations changed in the comparison with the control conditions approaching the male actors' values, which were different from those of the female participants and actors. The variation in lip kinematics induced changes also in voice formants but only in the visual presentation. The data suggest that both features of the lip kinematics and of the voice spectra tend to be automatically imitated when repeating a string-of-phonemes presented by a visible and/or audible speaking interlocutor. The use of imitation, in place of the usual lip kinematics and vocal features, suggests an automatic and unconscious tendency of the perceiver to interact closely with the interlocutor. This is in accordance with the idea that resonant circuits are activated by the activity of the mirror system, which relates observation to execution of arm and mouth gestures. PMID- 16698052 TI - Physical abuse during pregnancy and risk of low-birthweight infants among aborigines in Taiwan. AB - OBJECTIVES: The aim of this study was to assess whether physical abuse during pregnancy is associated with an increased risk of giving birth to a low birthweight (LBW) infant. STUDY DESIGN AND METHODS: A cross-sectional survey was conducted and 1143 aboriginal women were recruited into this study. The Abuse Assessment Screen was used to collect information regarding maternal physical abuse, and infants' birth weights were obtained from hospital medical records. Multiple logistic regression was used to estimate the association of LBW with physical abuse during pregnancy, adjusting for behavioural and sociodemographic variables. RESULTS: Of the women experiencing physical abuse during their recent pregnancy, 11.76% delivered a LBW infant, compared with 5.78% of women who did not experience physical abuse [odds ratio (OR) 1.97, 95% confidence intervals (CI) 1.07-3.63]. Physical abuse during pregnancy was also significantly associated with low weight gain in the prenatal period. Univariate analysis revealed that delivery of a LBW infant was associated with maternal alcohol use, maternal betel chewing and maternal educational level. Multiple logistic regression was performed, adjusting for prenatal weight gain, maternal height, maternal years of education, maternal alcohol use and maternal betel quid use. It was found that women who reported physical abuse with injuries during pregnancy had a 2.4-fold higher risk of delivering a LBW infant compared with women who did not report physical abuse with injuries (adjusted OR=2.43, 95%CI 1.06-5.55). CONCLUSIONS: These findings suggest that physical abuse during pregnancy is an independent risk factor for delivery of a LBW infant. Prenatal care for aboriginal women should be coupled with routine systematic screening for the presence of abuse during pregnancy, and adequate support and interventions for abused women. PMID- 16698053 TI - Efficacy and safety of two whole IgG polyvalent antivenoms, refined by caprylic acid fractionation with or without beta-propiolactone, in the treatment of Bothrops asper bites in Colombia. AB - The efficacy and safety of two whole IgG polyvalent antivenoms (A and B) were compared in a randomised, blinded clinical trial in 67 patients systemically envenomed by Bothrops asper in Colombia. Both antivenoms were fractionated by caprylic acid precipitation and had similar neutralising potencies, protein concentrations and aggregate contents. Antivenom B was additionally treated with beta-propiolactone to lower its anticomplementary activity. Analysing all treatment regimens together, there were no significant differences between the two antivenoms (A=34 patients; B=33 patients) in the time taken to reverse venom induced bleeding and coagulopathy, to restore physiological fibrinogen concentrations and to clear serum venom antigenaemia. Blood coagulability was restored within 6-24 h in 97% of patients, all of whom had normal coagulation and plasma fibrinogen levels 48 h after the start of antivenom treatment. Two patients (3.0%) had recurrent coagulopathy and eight patients suffered recurrence of antigenaemia within 72 h of treatment. None of the dosage regimens of either antivenom used guaranteed resolution of venom-induced coagulopathy within 6 h, nor did they prevent recurrences. A further dose of antivenom at 6 h also did not guarantee resolution of coagulopathy within 12-24 h in all patients. The incidence of early adverse reactions (all mild) was similar for both antivenoms (15% and 24%; P>0.05). PMID- 16698054 TI - Prevalence of Clostridium difficile in the environment in a rural community in Zimbabwe. AB - Clostridium difficile has been shown to be a nosocomial pathogen associated with diarrhoea and pseudomembranous colitis in hospitalised patients, but very little is known about its prevalence outside the hospital environment. The aim of this study was to determine the prevalence of C. difficile in faeces of domestic animals, soil and drinking water in a rural community. Water, animal faeces and soil were collected from homesteads in a rural community and the samples were cultured for C. difficile. Clostridium difficile isolates that produced toxins A or B were tested for their susceptibility to antimicrobial drugs. Clostridium difficile was isolated from 37.0% of 146 soil samples, 17.4% of 115 chicken faeces samples, 6.0% of 234 water samples and 4.3% of 161 faecal samples of other animals. Some of the C. difficile isolates from chickens (55.0%), soil (66.7%) and water (14.3%) were toxigenic. All toxigenic isolates were susceptible to metronidazole, vancomycin, doxycycline, chloramphenicol and tetracycline and all were resistant to cefotaxime, gentamicin, ciprofloxacin, norfloxacin and nalidixic acid. The results of the present study suggest that chickens kept by villagers are an important reservoir of C. difficile, which may act as a source of human infection. PMID- 16698055 TI - Volatiles from the venom of five species of paper wasps (Polistes dominulus, P. gallicus, P. nimphus, P. sulcifer and P. olivaceus). AB - The venom volatiles of five paper wasp species, four European belonging to the subgenus Polistes sensu stricto (P. dominulus, P. gallicus, P. nimphus, P. sulcifer) and one belonging to the Asian subgenus Gyrostoma (P. olivaceus), have been sampled by headspace solid phase micro-extraction and analysed by gas chromatography-mass spectrometry. The venom volatile components of Polistes wasps have never been fully investigated before, although the presence of some spiroacetals has been previously reported in literature. The composition of the venom was qualitatively and quantitatively different among the analysed species with the major substances tentatively identified, on the basis of their mass spectra, as: spiroacetals, mainly 2,8-dimethyl-1,7-dioxaspiro[5.5]undecane, two amides, N-(3-methylbutyl)acetamide and N-(3-methylbutyl)propanamide and acetates of saturated, mono- and di-unsaturated 2-alcohols with an odd number of carbon atoms in the chain. The acetate of a di-unsaturated 2-alcohol, present in two isomeric forms, identified as (E)- and (Z)-5-tangerinol has never been reported in literature for insects. Propanoates of the same 2-alcohols were only found in the venom of P. gallicus. Both the amides and the above-mentioned spiroacetal have been already shown to be alarm pheromones in other social wasps, while the acetates and propanoates have ever been reported in this taxon. PMID- 16698056 TI - Slant perception, and its voluntary control, do not govern the slant aftereffect: multiple slant signals adapt independently. AB - Although it is known that high-level spatial attention affects adaptation for a variety of stimulus features (including binocular disparity), the influence of voluntary attentional control-and the associated awareness-on adaptation has remained unexplored. We developed an ambiguous surface slant adaptation stimulus with conflicting monocular and binocular slant signals that instigated two mutually exclusive surface percepts with opposite slants. Using intermittent stimulus removal, subjects were able to voluntarily select one of the two rivaling slant percepts for extended adaptation periods, enabling us to dissociate slant adaptation due to awareness from stimulus-induced slant adaptation. We found that slant aftereffects (SAE) for monocular and binocular test patterns had opposite signs when measured simultaneously. There was no significant influence of voluntarily controlled perceptual state during adaptation on SAEs of monocular or binocular signals. In addition, the magnitude of the binocular SAE did not correlate with the magnitude of perceived slant. Using adaptation to one slant cue, and testing with the other cue, we demonstrated that multiple slant signals adapt independently. We conclude that slant adaptation occurs before the level of slant awareness. Our findings place the site of stereoscopic slant adaptation after disparity and eye posture are interpreted for slant [as demonstrated by Berends et al. (Berends, E. M., Liu, B., & Schor, C. M. (2005). Stereo-slant adaptation is high level and does not involve disparity coding. Journal of Vision 5 (1), 71-80), using that disparity scales with distance], but before other slant signals are integrated for the resulting awareness of the presented slant stimulus. PMID- 16698057 TI - Amplitude adaptation occurs where a saccade is represented as a vector and not as its components. AB - When saccades become inaccurate, their amplitude is adapted. We examined, in humans, whether this adaptation occurs where the saccade is represented as a vector or as its horizontal and vertical components. In one experiment, we behaviorally reduced the amplitude of clockwise oblique saccades and examined the transfer to saccades made to other target amplitudes and directions. In a second, we adapted rightward saccades of the same size as the rightward component of the clockwise oblique saccades and examined the effect on oblique saccades. The results of both experiments imply that adaptation occurs where the saccade command is represented as a vector. PMID- 16698058 TI - With a careful look: still no low-level confound to face pop-out. AB - In this issue of Vision Research, VanRullen, R. (2006). On second glance: Still no high-level pop-out effect for faces. Vision Research, in press. challenges our earlier Vision Research paper, "At first sight: A high-level pop-out effect for faces" (Hershler, O., & Hochstein, S. (2005). At first sight: A high-level pop out effect for faces. Vision Research, 45, 1707-1724). In that paper, we showed that faces pop-out from a great variety of heterogeneous distractors. This search must have been based on a holistic combination of facial features, since it could not have relied on any single low-level distinguishing feature-each of which was present in at least some of the distractors. VanRullen implies that the pop-out effect is not limited to faces, is not holistic, and is due to a low-level confound, namely that the "low-level" Fourier amplitude spectrum may differentiate between faces and other categories. We now show that he fails to substantiate all three claims. His first experiment replicates our own and shows once again that faces do indeed pop-out, while other objects, such as cars, do not. The claim regarding the non-holistic nature of face search is based on a failure to differentiate between holistic processing for face detection and for individual face identification. His central claim is that the Fourier amplitude spectrum is processed low-level and could be used for face pop-out. However, changing the amplitude spectrum may well affect high-level representations as well. For example, his demonstration uses hybrid images which are extremely fuzzy, rendering them difficult to identify. More importantly, this claim would lead to the conclusion that targets with a non-face phase spectrum and only a face amplitude spectrum would pop-out among distractors with different amplitude spectra. We demonstrate that this is, of course, not the case and that the Fourier amplitude is not the hoped for "low-level confound". Until another such "hidden" low level feature is found, we must accept that face pop out depends on a high level mechanism. PMID- 16698059 TI - Interaction of spatial and temporal integration in global form processing. AB - The mechanisms by which global structure is extracted from local orientation information are not well understood. Sensitivity to global structure can be investigated using coherence thresholds for detection of global forms of varying complexity, such as parallel and concentric arrays of oriented line elements. In this study, we investigated temporal integration in the detection of these forms and its interaction with spatial integration. We find that for concentric patterns, integration times drop as region size increases from 3 degrees to 10.9 degrees , while for parallel patterns, the reverse is true. The same spatiotemporal relationship was found for Glass patterns as for line element arrays. The two types of organization therefore show quite different spatiotemporal relations, supporting previous arguments that different types of neural mechanism underlie their detection. PMID- 16698060 TI - Effects of feature-based attention on the motion aftereffect at remote locations. AB - Previous studies have shown that attention to a particular stimulus feature, such as direction of motion or color, enhances neuronal responses to unattended stimuli sharing that feature. We studied this effect psychophysically by measuring the strength of the motion aftereffect (MAE) induced by an unattended stimulus when attention was directed to one of two overlapping fields of moving dots in a different spatial location. When attention was directed to the same direction of motion as the unattended stimulus, the unattended stimulus induced a stronger MAE than when attention was directed to the opposite direction. Also, when the unattended location contained either uncorrelated motion or had no stimulus at all an MAE was induced in the opposite direction to the attended direction of motion. The strength of the MAE was similar regardless of whether subjects attended to the speed or luminance of the attended dots. These results provide further support for a global feature-based mechanism of attention, and show that the effect spreads across all features of an attended object, and to all locations of visual space. PMID- 16698061 TI - Chlorination kinetics of glyphosate and its by-products: modeling approach. AB - Chlorination reactions of glyphosate, glycine, and sodium cyanate were conducted in well-agitated reactors to generate experimental kinetic measurements for the simulation of chlorination kinetics under the conditions of industrial water purification plants. The contribution of different by-products to the overall degradation of glyphosate during chlorination has been identified. The kinetic rate constants for the chlorination of glyphosate and its main degradation products were either obtained by calculation according to experimental data or taken from published literature. The fit of the kinetic constants with experimental data allowed us to predict consistently the concentration of the majority of the transitory and terminal chlorination products identified in the course of the glyphosate chlorination process. The simulation results conducted at varying aqueous chlorine/glyphosate molar ratios have shown that glyphosate is expected to degrade in fraction of a second under industrial aqueous chlorination conditions. Glyphosate chlorination products are not stable under the conditions of drinking water chlorination and are degraded to small molecules common to the degradation of amino acids and other naturally occurring substances in raw water. The kinetic studies of the chlorination reaction of glyphosate, together with calculations based on kinetic modeling in conditions close to those at real water treatment plants, confirm the reaction mechanism that we have previously suggested for glyphosate chlorination. PMID- 16698062 TI - Homologue and isomer distribution of dioxins observed in water samples collected from Kahokugata Lagoon and inflowing rivers, Japan. AB - Water samples were collected at 17 sites in Kahokugata Water Basin, a closed water basin in central Japan. We determined the concentration of dioxins of the water samples. Linear relationships between toxic equivalent (TEQ) concentrations of dioxin and concentrations of suspended solid (SS) were obtained at sites in Kahokugata Lagoon and in the rivers flowing into the lagoon. Homologue composition of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) indicated that all the water samples were still strongly influenced by chlorinated herbicides, such as chloronitrofen (CNP) and pentachlorophenol (PCP) that had been widely used in rice fields. The main isomer distributions of the PCDD homologues were not significantly different among the sampling sites, while the main isomer distributions of the PCDF homologues were considerably different among the sampling sites. At a few sampling points in the downstream part of one of the rivers, high concentrations of 1,3,6,7,8-pentachloro dibenzofuran (1,3,6,7,8-PeCDF) and its related isomers (1,3,6,8-chlorine-substituted PCDFs) were traced to a dye manufacturing plant. These non-toxic isomers are usually only minor constituents in environmental water samples and are not indicators of any known dioxin sources. The dyeing discharge was found to make a contribution only in the water samples collected near the plant and the seasonal variation of the contribution might depend on the flow rate of the river. PMID- 16698063 TI - Comparative assessment of coal tars obtained from 10 former manufactured gas plant sites in the eastern United States. AB - A comparative analysis was performed on eleven coal tars obtained from former manufactured gas plant sites in the eastern United States. Bulk properties analyzed included percent ash, Karl Fisher water content, viscosity and average molecular weight. Chemical properties included monocyclic- and polycyclic aromatic hydrocarbon (PAH) concentrations, alkylated aromatic concentrations, and concentrations of aliphatic and aromatic fractions. It was found that there was at least an order-of-magnitude variation in all properties measured between the eleven coal tars. Additionally, two coal tars obtained from the same manufactured gas plant site had very different properties, highlighting that there can be wide variations in coal tar properties from different samples obtained from the same site. Similarities were also observed between the coal tars. The relative chemical distributions were similar for all coal tars, and the coal tars predominantly consisted of PAHs, with naphthalene being the single-most prevalent compound. The C(9-22) aromatic fraction, an indicator of all PAHs up to a molecular weight of approximately 276 gmole(-1), showed a strong power-law relationship with the coal tar average molecular weight (MW (ct)). And the concentrations of individual PAHs decreased linearly as MW (ct) increased up to ca. 1000 gmole(-1), above which they remained low and variable. Implications of these properties and their variation with MW (ct) on groundwater quality are discussed. Ultimately, while these similarities do allow generalities to be made about coal tars, the wide range of coal tar bulk and chemical properties reported here highlights the complex nature of coal tars. PMID- 16698064 TI - The degradation of the cyanobacterial hepatotoxin nodularin (NOD) by UV radiation. AB - This study investigates the decomposition of NOD by UV irradiation. Water solutions of pure NOD and NOD-containing Nodularia extract as well as Nodularia filaments collected on filters were exposed to UV-A, UV-B, and white fluorescent light (VIS) during 48 h experiments. In VIS, the toxin was fairly stable and only 3.8-4.6% of the original degraded. UV-B had the most pronounced effect on the NOD degradation rate. In the experiment, the overall loss of NOD was 0.27 and 0.77 micro g ml(-1)day(-1) for the solution of pure toxin and Nodularia extract and 0.28 micro g day(-1) for Nodularia filaments. Comparison of UV-B degradation rate in water and methanol extracts revealed higher stability of NOD in methanol. This might suggest that some hydrophobic components of Nodularia cell play a protective role against UV radiation. Additionally, chemical (LC-MS/MS) and biochemical (ELISA and PPIA) assays were employed to characterize the UV degradation products. LC-MS/MS analyses showed that in UV-B exposed sample, apart from NOD, there were three other compounds with molecular ion at m/z at 825.4. The fragmentation pattern of the ion was the same for all four compounds suggesting that they are geometrical isomers of NOD. The major degradation product, with a local absorption maximum at 242 nm, was active in both biochemical assays. PMID- 16698065 TI - Compost effect on soil humic acid: A NMR study. AB - The humic acid (HA) fraction of a food and vegetable residues compost (CM) was taken as indicator to trace the fate of CM organic matter in four years CM amended soil. (1)H and (13)C NMR spectroscopy were used to investigate the nature of the HA isolates from CM, control soil (S(4)) and amended soil. The result indicated a significant structural difference between CM HA and S(4) HA, and supported the presence of both HA fractions in soil at the end of the amendment trials. However, the nature and content of CM HA in soil did not fully explain the increase of soil cation exchange capacity (CEC) after amendment. All CM humic fractions (i.e., fulvic acid, humic acid and humin) were found to contribute to the change of the soil organic matter composition. It is concluded that although CM HA is a suitable indicator of the survival of compost organic matter in soil during amendment, all three humic fractions should be monitored and analyzed to fully understand changes in the composition and properties of amended soil. PMID- 16698066 TI - Characterization and modeling of nutrient-deficient tomato-processing wastewater treatment using an anaerobic/aerobic system. AB - Tomato-processing wastewaters are typical of slowly biodegradable high strength wastewaters generated from the food canning industry. Three different samples of influent and three samples of anaerobic effluents corresponding to the three influents collected from an on-site pilot-scale anaerobic/aerobic system were characterized using respirometric methods. Respirometric studies demonstrated that the pretreatment in the anaerobic reactor of the pilot-scale system increased the readily biodegradable fraction from an average of 40% of the SCOD in the influent to 50% of the SCOD in the anaerobic effluent, improved kinetics from an average micro(max) of 1.5d(-1) in the influent to 3.5d(-1) in the anaerobic effluent, and eliminated nutrient deficiency. Furthermore, the system was calibrated and simulated by application of wastewater characterization and biokinetic data derived form respirometric tests of influent and anaerobic effluent in Activated Sludge Model no.1 (ASM1). PMID- 16698067 TI - Effect of crude oil contaminated sediment exposure on cytochrome P450 enzymes in the Australian asteroid Coscinasterias muricata. AB - Levels of cytochrome P450 enzymes were measured in pyloric caeca microsomes of the asteroid Coscinasterias muricata following exposure to sediment with nominal concentrations of 0, 0.1 or 2 ml crude oil kg(-1) (dry weight) and subsequent depuration. No significant differences were observed in total cytochrome P450 levels or cytochrome P418 levels following the exposure period. However after five days of depuration, levels of total P450 in the pyloric caeca of C. muricata exposed to the highest oiled sediment concentration were significantly lower than in specimens exposed to the other treatments. Cytochrome P418 levels were inversely related to total P450 levels following exposure and subsequent depuration. Preliminary results show that levels of CYP1A-like immunopositive protein (CYP1A-like IPP) in exposed asteroids exhibited a concentration response relationship following the exposure period. Variations in CYP1A-like IPP levels observed during the depuration period may be influenced by the sublethal toxicity of hydrocarbons within the crude oil. PMID- 16698068 TI - Ethoxyresorufin-O-deethylase (EROD) activity and bile metabolites as contamination indicators in Baltic Sea perch: determination by HPLC. AB - Exposure of Baltic Sea perch (Perca fluviatilis) to organic contaminants was investigated by means of liver enzyme activity, measured as ethoxyresorufin-O deethylase (EROD) activity, and PAH metabolites excreted to bile. Female perch used in the study were caught in coastal waters of the western Gulf of Finland. Reaction conditions were optimised for determination of EROD activity in liver. A high-performance liquid chromatography (HPLC) method with fluorescence detection was developed to separate the interfering fluorescent signal from the signal due to EROD activity. EROD activity in perch varied within a concentration range of 0.30-14 pmol min(-1)mg(-1) protein. Recent PAH exposure was indicated in enhanced levels of the bile metabolite 1-hydroxypyrene, which varied between 213 and 1149 microg kg(-1). No correlation was indicated between hepatic EROD activity and concentration of 1-hydroxypyrene. PMID- 16698069 TI - Lead isotope ratio measurements by ICP-QMS to identify metal accumulation in vegetation specimens growing in mining environments. AB - The use of variations in stable Pb isotope ratios has become a well-established diagnostic technique for characterising sources of lead contamination. In this work, lead isotope ratios in mining wastes (lead content 320-130,000 mg kg-1) and vegetation specimens (lead concentration 7-650 mg kg-1) have been determined by inductively coupled plasma quadrupole-based mass spectrometry (ICP-QMS) in order to investigate lead bioaccumulation in Buddleia davidii growing on wastes from two abandoned Pb/Zn mining areas in Spain. The accuracy of the isotope ratio measurements was evaluated by analysing a certified isotopic standard NIST SRM 981. Good agreements were obtained between the lead isotope ratios measured and the certified values (deviations within 0.01-0.2%). The results indicate that the lead isotopic ratios in vegetation samples collected in the mining areas differed from those of a specimen from an uncontaminated site (control sample). However, close lead isotope ratio values were found between vegetation specimens and mining tailings. Therefore, the results suggest that lead in the collected vegetation specimens is most likely related to the influence of mining activities rather than to other sources like past leaded-petrol emissions. PMID- 16698070 TI - Sediment reference concentrations of seldom monitored trace elements (Ag, Be, In, Ga, Sb, Tl) in four Swedish boreal lakes--comparison with commonly monitored elements. AB - This paper presents reference and recent acid-leachable concentrations of some seldom monitored trace elements (SMTE; Ag, Be, Ga, In, Sb and Tl) in sediments from four boreal oligotrophic lakes in a south to north transect in Sweden. For comparison commonly monitored trace elements (As, Cd, Co, Cr, Cu, Ni, Pb and Zn) are included as well as those of relevance for redistribution processes (Al, Ca, Fe, Mg and Mn). Pore water pH and the corresponding solid/solution distribution coefficients (Kd) indicated that redistribution of the elements due to acidification is minor. The depth of impact was defined as the level in the sediment where the mean values became constant on successive exclusion of metal concentrations in overlying strata. Reference concentrations were calculated below the depth of impact. The present concentration changes are estimated by the ratio between the average concentration above the depth of impact and the reference concentration. Reference concentration ranges for the SMT-elements are (mg/kg, dry wt.): Ag 0.16-0.66; Be 1.6-3.7; Ga 2.0-5.1; In 0.05-0.22; Sb 0.05 0.11 and Tl 0.17-0.70. The concentration ratios for these elements ranged in the two most southern lakes from 1.5 to 4.5 and in the two northern ones from 0.6 to 1.6. A high correlation between Kd for the SMT-elements, and iron, except for Sb and Tl, infer that the biogeochemistry of iron is quantitatively important for the accumulation of these elements. The reference concentration ranges for the commonly monitored trace elements are (mg/kg, dry wt.): As 7.0-29.6; Cd 0.33 0.98; Co 5.7-23.8; Cr 15.2-26.1; Cu 27.6-58.4; Ni 5.4-20.8; Pb 44-96. The corresponding concentration ratios were 1.2-18 (second highest 3.9) in the two most southern lakes and 0.8-1.6 in the two northern ones. Declining ratios were found from south to north, most obvious for cobalt and zinc. The copper ratios did not show a regional pattern, partly because of the impact from old mine waste. Increased concentrations of Ag, Be, Ga, In, Sb and Tl in recent sediments up to 4.5 times the reference levels in combination with the geographical pattern infer an elevated loading of these elements. PMID- 16698071 TI - Age-dependent morphological alterations of human ovaries from populations with and without BRCA mutations. AB - OBJECTIVE: From analysis of pre-cancer ovarian tissues obtained from prophylactic oophorectomies, several studies reported the increased ovarian morphological changes in high-risk ovaries, but whether these morphological changes are associated with BRCA1/BRCA2 genotypes or are cancer precursors is controversial. Here, we have investigated a recent collection of ovaries from prophylactic oophorectomies and control ovaries from surgeries due to other non-ovarian related cancer or non-neoplastic diseases to determine if ovarian morphological changes relate to BRCA1/2 genotypes or reproductive history. METHODS: We assembled a panel of archived ovarian tissues: 52 ovarian tissue blocks were from prophylactic oophorectomies of a high-risk (HR) population; 66 ovaries were from surgeries due to non-ovarian-related diseases, referred to as normal-risk (NR) group. The morphology of ovarian tissues was examined, and morphological changes including papillomatosis, invaginations, inclusion cysts, and epithelial stratification were assessed in a blinded fashion. RESULTS: No statistically significant difference in frequency of these histolopathologic features was found between HR and NR groups. However, inclusion cysts and deep invaginations were found much more commonly in women age 45-54 of either HR or NR groups. CONCLUSIONS: This study suggests that no significant increase in the presence of non-neoplastic ovarian morphological changes is associated with BRCA1/BRCA2 mutations. Rather, the frequency of these histological features, especially inclusion cysts, may associate with age or menopausal status. We propose that ovulatory and perimenopausal gonadotropin stimulation produces ovarian morphological changes, and these histological features may promote the transformation of genetically compromised epithelial cells in the development of ovarian cancer. PMID- 16698072 TI - Effectiveness and dentin bond strengths of two materials for reinforcing thin walled roots. AB - OBJECTIVES: To investigate the effectiveness of two restorative materials for reinforcing thin-walled roots and their microtensile bond strengths to root canal dentin. METHODS: Twenty-one decoronated maxillary central incisor roots were root filled and the canals enlarged to leave approximately 1.0 mm thick dentin walls. The roots were distributed randomly to three equal groups. Group 1 (control): a large tapered cast post-core was fabricated. Group 2 (BIS-CORE): a thick layer of dual-cured composite was placed in the post-hole before fabrication of a small diameter tapered cast post-core. Group 3 (ChemFil Superior): a thick layer of glass-ionomer was placed, as before. Metal-ceramic crowns were fabricated for all teeth. Microtensile bond strength tests and optical and atomic force microscope observations were employed to examine adhesion between the two restorative materials and the root canal dentin. RESULTS: Mechanical loading (kN) demonstrated that composite 0.64 (S.D. 0.05), but not glass-ionomer 0.49 (0.05), significantly reinforced the fracture resistance of thin-walled roots, P<0.001. Microtensile bond strengths (MPa) were 21.7 (1.6) for composite and 12.9 (1.7) for glass-ionomer, P=002. SIGNIFICANCE: A thick intermediate layer of resin bonded composite, sandwiched between the root dentin and a small-diameter cast Ni Cr post or dowel, increased significantly the fracture resistance of the roots. PMID- 16698073 TI - The role of ras gene in the development of haemic neoplasia in Mytilus trossulus. AB - Disseminated neoplasia has been reported in mussels (Mytilus spp) from numerous locations worldwide. This condition is progressive and fatal and the aetiology is unknown. In vertebrates, oncogenes such as ras, and tumour suppressor genes such as p53, play important roles in carcinogenesis. We have cloned a Mytilus trossulus homologue of the vertebrate ras gene, which shows conserved sequence in regions of functional importance. Neoplastic hemolymph samples derived from M. trossulus have been investigated for the presence of ras gene mutations and changes in expression. PMID- 16698074 TI - Oxidative damage produced by Cr(VI) and repair in mussel (Mytilus edulis L.) gill. AB - This study has assessed DNA damage induced by oxidative stress and its subsequent repair in mussels. Gill was obtained from mussels collected from New Brighton, UK within 24 h and also after 1 month maintenance under laboratory conditions. The pro-oxidant sodium dichromate produced a statistically significant increase in DNA strand breaks (DSB) in these gill cells at both time points as measured by the COMET assay. The response was higher at 1 month in association with a higher concentration of GSH which is known to activate Cr(VI) producing reactive oxygen species. DSB were shown, through studies in wild type and OGG-1-null mouse fibroblasts, to be produced by repair enzymes in response to Cr(VI). In support of evidence for repair of oxidative DNA damage, we have also demonstrated for the first time repair activity in mussel gill towards 8-oxo-dG using an oligonucleotide cutting assay. PMID- 16698075 TI - Benzo[a]pyrene cytotoxicity in right whale (Eubalaena glacialis) skin, testis and lung cell lines. AB - The North Atlantic right whale (NARW) is one of the most endangered great whales. The NARW population consists of only about 300 individuals and is reproducing at an insufficient rate. There is growing concern about the potential effects of environmental contaminants on the reproductive and overall health of NARW. High contaminant burdens can accumulate in tissues of great whales but toxicological studies of their effects are limited due to legal, logistical and ethical restrictions and specific in vitro models are critically needed. Cell lines from NARW skin and internal organs were previously created in our laboratory. In this study, skin, testis and lung primary fibroblast cell lines were exposed to benzo[a]pyrene (BP) as part of a multi-chemical toxicity testing project in NARW. Cells were exposed for 24-72 h to 10 nM-10 microM BP dissolved in dimethylsulfoxide. Cytotoxicity was measured with a clonogenic assay using standard methods. Some cytotoxicity was observed after 24 h, the highest concentration (10 microM BP) resulting in 77, 74 and 51 percent relative survival in testis, skin and lung cells, respectively, and indicating a higher cytotoxicity in the lung (p < 0.05). After 48 and 72-h exposure, 10 microM BP resulted in 24 and 3, 74 and 27, and 42 and 23 percent relative survival in testis, skin and lung cells, respectively. Cytotoxicity significantly increased with exposure time in all three tissues (p < 0.05 for skin and p < 0.01 for lung and testis), suggesting metabolic activation of BP in the three organs. Fibroblast cytotoxicity observed in the testis was higher than that observed either in the skin or lung after 48 h (p < 0.01) and was close to 100% after 72 h, warranting further investigation of the potential effects of PAHs on reproductive health. PMID- 16698076 TI - Aromatase in zebrafish: a potential target for endocrine disrupting chemicals. AB - In zebrafish, two isoforms of the aromatase gene exist, namely cyp19a1 and cyp19a2, expressed predominantly in the gonads and brain, respectively. In this study, we focus on characterizing the specificity of antibodies against the aromatase isoforms, and on (xeno)estrogen-induced changes of individual cyp19a2 mRNA concentrations in the brains of adult male zebrafish. Among three polyclonal antibodies studied, the one against CYP19A2 was found to be specific in Western blots and immunohistochemistry. Real-time RT-PCR analyses revealed strong interindividual variation of cyp19a2 levels in the brains of adult male zebrafish. After a three-week-exposure to (xeno)estrogens, mean values of cyp19a2 mRNA levels tended to increase, with significant induction at 200 ng 17beta estradiol/L, but interindividual variation of cyp19a2 expression was maintained. PMID- 16698077 TI - Injectable calcium phosphate scaffold and bone marrow graft for bone reconstruction in irradiated areas: an experimental study in rats. AB - The purpose of this study was to assess the possibilities for bone reconstruction of an injectable calcium phosphate scaffold (ICPS) associated with a bone marrow (BM) graft after irradiation in a rat model. External irradiation was delivered to 12 out of 27 inbred rats. Three weeks later, four osseous defects were created per animal and were kept empty or filled with either ICPS alone, BM graft alone or with a mixture of BM and ICPS. Three weeks after implantation, bone specimens were studied under light microscopy and by scanning electron microscopy. Filling irradiated defects with ICPS alone was not accompanied by the formation of new bone. The BM graft associated with ICPS significantly increased ceramic degradation (p<0.01) and bone ingrowth (p<0.01) in the irradiated areas. The results are evidence for the meaning of the BM in driving the bone repair in irradiated animals. PMID- 16698078 TI - Expression of urocortin in the extravillous human trophoblast at the implantation site. AB - Urocortin (UCN) is a 40 amino acid peptide which is closely related to corticotropin-releasing hormone and binds with high affinity to both CRH type 1 and type 2 receptors. UCN is expressed in human reproductive tissues including endometrium, ovary, and placenta. This study was designed to investigate the cellular localization of UCN at the implantation site of the human blastocyst, as well as the regulation of the UCN promoter by two major intracellular signaling pathways, the cAMP/PKA and diacylglycerol/PKC pathways, in cells of placental origin. For this reason, immunohistochemistry was performed on tissue sections from paraffin-embedded human first trimester placentas and freshly isolated human invasive extravillous trophoblast cells (EVT) were analyzed for UCN expression using RT-PCR and immunofluorescence. Finally, UCN promoter activity was analyzed in the JEG3 human choriocarcinoma cell line. Immunohistochemistry revealed expression of UCN in the cytotrophoblast, the EVT and decidual cells. Both UCN mRNA and peptide were detectable in freshly isolated EVT. Finally, a human UCN promoter luciferase reporter construct transfected into JEG3 cells was significantly inducible by phorbol ester plus ionomycin, but not by phorbol ester alone or by forskolin. Collectively, the present study reports the expression of UCN in EVT and the activation of the UCN gene promoter by the diacylglycerol/PKC pathway. The functional significance of urocortin for the physiology of EVT requires further investigation. PMID- 16698079 TI - Increased chymotrypsin-like protease (chymase) expression and activity in placentas from women with preeclampsia. AB - Placenta-derived chymotrypsin-like protease (CLP/chymase) promotes endothelial P selectin and E-selectin expression, which may be responsible for the increased neutrophil/endothelial interactions in preeclampsia (PE). However, little is known about this protease expression and production in human placenta. This study was undertaken to determine the distribution and gene expression of CLP in human placenta. Human placental tissues were obtained immediately after delivery from normal and PE pregnancies. We examined (1) CLP/chymase immunoactivity by immunohistochemical staining of villous tissue sections; (2) trophoblast mRNA and protein expression for chymase by RT-PCR and Western blot analysis; (3) chymase cDNA sequencing in isolated trophoblast cells (TCs); and (4) release of CLP by placental villous tissue cultured under 2% and 20% O(2). We found (1) CLP expression is mainly localized in the epithelial layer of syncytiotrophoblasts; (2) both mRNA and protein expression are significantly (p<0.05) upregulated in TCs isolated from PE vs. normal placentas; (3) TC chymase cDNA sequence and the deduced amino acid sequence are 100% identical to that reported for the human heart; and (4) villous tissue releases more chymotrypsin when cultured with 2% O(2). We conclude that (1) the DNA and protein sequence for chymase in placental trophoblast cells are the same as those reported in the human heart; (2) CLP/chymase expression is upregulated in TCs during PE; and (3) lowered oxygen condition promotes CLP release by placental TCs. Since chymase is a potent non ACE angiotensin II producing enzyme, our data suggest that if placenta-derived CLP/chymase is released into the maternal circulation, it may contribute to the cardiovascular complications associated with PE. PMID- 16698080 TI - Comparing maltreating fathers and mothers in terms of personal distress, interpersonal functioning, and perceptions of family climate. AB - OBJECTIVE: This study compared perceptions of personal distress, interpersonal and marital problems, and aspects of family climate of maltreating fathers and mothers. METHODS: Subjects were 2841 offenders (1918 of whom were fathers or father-figures) who were identified and treated by the USAF Family Advocacy Program between 1988 and 1996. Independent variables for the analysis were parent sex (mother vs. father) as well as type and severity of maltreatment, history of repeat offenses, and history of abuse in childhood. RESULTS: Maltreating mothers were more distressed and reported more problems from individuals outside the family than maltreating fathers; fathers reported more rigid expectations for children, less cohesive families, and less organized families than did maltreating mothers. Regardless of parental sex, victimization in the family of origin was related to distress and unhappiness. Similarly, both victimization in the family of origin and history of repeated offenses were powerful predictors of a more negative family climate regardless of the offending parent's sex. No significant statistical interactions between parental sex and other independent variables were found when predicting personal and interpersonal distress, marital problems, or family climate. CONCLUSIONS: Studies rarely examine maltreating fathers except in the context of sexual abuse. Fewer still compare maltreating mothers and fathers. This study identified meaningful, though generally small, differences between maltreating mothers and fathers. Patterns suggest that maltreating mothers may tend to cope more poorly with personal distress, whereas maltreating fathers tend to operate in a family climate that is both distant and rigid, while holding inappropriate expectations for children's behavior. The absence of interactions between parental sex and the other independent variables included in the analysis indicate that these patterns do not vary by the history of victimization in the family of origin, the type or severity of child maltreatment, or the history of prior maltreatment in the family. PMID- 16698081 TI - How useful is the presence of petechiae in distinguishing non-accidental from accidental injury? PMID- 16698087 TI - A novel stimulus system for applying tactile stimuli to the macrovibrissae in electrophysiological experiments. AB - The rat's vibrissae are a wonderful system for the study of sensory neural encoding in relation to behaviour because the vibrissae are easily identifiable and accessible for manipulation, allowing easy application of a variety of different types of deflections that mimic natural whisking. Here we report the development of a powerful and flexible method for precisely deflecting these vibrissae. Recordings from CNS neurons showed, in response to variations in the parameters of a trapezoid whisker deflection stimulus that mimics the basic unit of whisking, a variety of complex responses as well as complex interactions between different response components. The recordings also included a response that is reported to be found during active whisking (movement under muscle control) and not passive whisker movements and thus to differentiate active from passive whisker deflections. Thus, this system could well be used in anaesthetized animals to apply whisker deflections that well mimic natural active whisking in awake animals, thereby allowing highly detailed study of the neuronal responses and neuronal interactions found with natural whisking behaviour. PMID- 16698085 TI - Mycobacterium heckeshornense: an emerging pathogen responsible for a recurrent lung infection. AB - Mycobacterium heckeshornense was responsible for a severe, recurrent and chronic pulmonary infection in an immunocompetent 65-year-old woman. The pathogen, initially identified as Mycobacterium xenopi and considered as a contaminant, led to a delayed adapted antimicrobial treatment. Although M. heckeshornense is phenotypically closely related to M. xenopi, its pathogenicity is noticeably higher. Accurate molecular diagnosis methods and treatment guidelines are needed to improve the management of patients infected by this uncommon pathogen. PMID- 16698086 TI - A simple and rapid radiochemical choline acetyltransferase (ChAT) assay screening test. AB - A simple radiochemical choline acetyltransferase (ChAT) assay screening test was developed by measuring for [(3)H]acetylcholine ([(3)H]ACh) formed from 0.2 mM [(3)H]acetyl-coenzyme A ([(3)H]acetyl-CoA) and 1 mM choline by 0.2 mg of rat brain homogenates containing ChAT into 96-well microplates. A simple and rapid procedure for isolating [(3)H]ACh from the incubation mixture into 96-well microplates was achieved by using a sodium tetraphenylboron (Kalibor) solution (in ethyl acetate, 0.75%, w/v) and a hydrophobic liquid scintillator mixture (1:5, v/v, 0.2 mL) as an extraction solvent. The benefits of this radiochemical method using 96-well microplates are as follows: (1) this method is reliable and reproducible; (2) many samples can be examined at the same time by this method; (3) this method is economical and effective in reducing radioactive waste. The development of a new simple radiochemical ChAT assay screening test is the first stage of development of radiolabeled ChAT mapping agent. PMID- 16698082 TI - Spiny mice (Acomys cahirinus) do not respond to thymus-independent type 2 antigens. AB - Analysis of the immune system of spiny mice (Acomys cahirinus) has been limited. Originally grouped with Mus, Acomys has recently been placed closer to Meriones (gerbils). This study compared immunity in Acomys, Mus, and Meriones. Lymphocytes from all rodents examined proliferated in response to mitogen and superantigen stimulation. Only Mus T cells responded to anti-CD3 stimulation. Acomys, like Meriones, and Mus that express xid, did not respond to thymus-independent type 2 antigens. Flow cytometric analyses revealed that T cell-specific MAbs did not bind Acomys or Meriones lymphocytes. The B cell-specific anti-CD45R (B220) MAb detected all rodent B cells and revealed the absence of a CD45R(lo) subset in the peritoneal cavity of Acomys and Meriones. Bone marrow from Acomys and Meriones failed to reconstitute B cell function in SCID mice. Thus, in terms of immunity, Acomys appears to be more similar to Meriones than Mus. PMID- 16698083 TI - Role of complement in neurodegeneration and neuroinflammation. AB - The complement system provides an innate defence mechanism against pathogenic microorganisms. Although viewed for many years as an immune-privileged organ, the central nervous system contains many components of the immune system, including components of the complement system that are synthesized by astrocytes, microglia, and neurons. During the past two decades, a wide range of inflammatory markers, typically absent in the normal elderly population, have been reported in Alzheimer's disease brains. It is becoming evident that sustained brain inflammation might be an essential cofactor in Alzheimer disease and other neurodegenerative disorders such as Parkinson disease, dementia with Lewy bodies, Huntington's and prion diseases. The complement system may be useful in eliminating aggregated and toxic proteins associated with these neurological disorders and thus have a protective effect. However, an exaggerated or insufficient activation of the complement system can have deleterious effect through the activation of microglia, secretion of many proinflammatory cytokines, and generation of oxidative products. The role of complement-mediated inflammation in Alzheimer disease has drawn greater attention recently in view of new therapeutic advances made in the management of the disease. This review is meant to update the role of complement in Alzheimer's disease and other neurodegenerative disorders in view of recent vaccination and immunotherapeutic approaches. PMID- 16698089 TI - Flow cytometric expression of common antigens CD18/CD45 in blood from dogs with lymphoid malignancies: a semi-quantitative study. AB - Flow cytometry is useful to study lymphoid malignancies since it allows both immunophenotyping of neoplastic cells and quantification of antigen expression. CD18 and CD45 are commonly exposed membrane antigens with different levels of expression on blood leukocyte and neoplastic cells. The aim of this retrospective study was to semi-quantitatively evaluate the expression of CD18 and CD45 in dogs with different lymphoid malignancies with blood involvement and to compare results with those from healthy dogs and dogs with reactive diseases. Blood samples from 13 dogs with precursor lymphoid malignancies, 20 with mature neoplasms (either chronic lymphocytic leukaemia or lymphoma), of different immunophenotypes, were compared with 24 healthy dogs and 12 dogs with different reactive diseases. The median fluorescence intensity (MFI) for CD18 and CD45 was recorded on lymphoid and granulocytic populations using dual colour flow cytometry, and the ratio between MFI for lymphoid and granulocytic populations (L/N ratio) was calculated to compare the results obtained in different sessions using an internal control (granulocyte fluorescence intensity). Significant decreases in the L/N ratio were detected in neoplastic samples for both CD18 (either precursors or mature versus controls) and CD45 (either precursors or mature versus control), while using MFI only slight differences were detectable in CD45 between precursors and controls. Neoplastic cells often exhibited lower expression of the L/N ratio for CD18, and mainly for CD45, most likely due to a less mature pattern than normal cells and/or to an aberrant quantitative expression of surface antigen. Moreover, more than 50% of neoplastic lymphoid cells exhibited L/N ratios that were not within the values observed in controls for at least one antigen. Altered L/N ratios, in particular decreases of CD45, were mainly observed in precursor neoplasms and in T-cell neoplasms. Detection of altered expression of common antigens, and in particular a L/N ratio for CD45 lower than a value of 103% may be useful as a confirmation of pseudo-clonality thus helping in differentiating reactive and neoplastic lymphocyte expansions. PMID- 16698088 TI - Antibody response in sheep experimentally infected with different small ruminant lentivirus genotypes. AB - Two groups of sheep were experimentally infected by intratracheal route with two small ruminant lentivirus (SRLV) isolates belonging to different genotypes (It 561 genotype A3 and It-Pi1 genotype B2). Seroconversion was evaluated using recombinant homologous and heterologous matrix protein/capsid antigen fusion protein. Results clearly indicate that seroconversion against homologous antigen was detected well in advance as regards heterologous antigen in both groups, although the advantage of using homologous antigen was less evident in detecting seroconversion against the caprine arthritis encephalitis virus (CAEV)-like strain, compared with the maedi-visna virus (MVV)-like infection. Commercially available ELISAs detect CAEV-like seroconversion earlier than MVV-like infection suggesting a closer relationship between CAEV-like isolate and the antigen used in the latter ELISA tests. Seven recombinant subunits developed from matrix protein and capsid antigen of strain K1514 (prototype A1) were used to better define the antibody response in sheep infected with It-561 isolate. Two animals clearly reacted against type specific epitopes in the early stage of infection. This study highlights the relative insensitivity of gag encoded cross-reacting epitopes during the early stage of infection and suggests the development of novel diagnostic tests based on both genotype specific antigens. PMID- 16698084 TI - Selective hypoxia-cytotoxins based on vanadyl complexes with 3-aminoquinoxaline-2 carbonitrile-N1,N4-dioxide derivatives. AB - A new vanadyl complex with the formula VO(L1)2, where L1=3-amino-6(7) chloroquinoxaline-2-carbonitrile N(1), N(4)-dioxide, has been synthesized and characterized by elemental analyses, conductometry, fast atom bombardment mass spectroscopy (FAB-MS) and electronic, Fourier transform infrared (FTIR), Raman, nuclear magnetic resonance (NMR) and electron paramagnetic resonance (EPR) spectroscopies. Results were compared with those previously reported for analogous vanadium complexes with other 3-aminoquinoxaline-2-carbonitrile N1,N4 dioxide derivatives as ligands. As an effort to develop novel metal-based selective hypoxia-cytotoxins and to improve bioavailability and pharmacological and toxicological properties of aminoquinoxaline carbonitrile N-dioxides bioreductive prodrugs, the new complex and VO(L)2 complexes, with L=3-amino-6(7) bromoquinoxaline-2-carbonitrile N1,N4-dioxide (L2) and 3-amino-6(7) methylquinoxaline-2-carbonitrile N1,N4-dioxide (L3), were subjected to cytotoxic evaluation in V79 cells in hypoxic and aerobic conditions. The complexes resulted in vitro more potent cytotoxins than the free ligands (i.e. potencies P(VO(L1)2)=3.0, P(L1)=9.0 microM) and Tirapazamine (P=30.0 microM) and showed excellent selective cytotoxicity in hypoxia, being no cytotoxic in oxia. In addition, the solubility in hydrophilic solvents resulted significantly higher for the vanadyl complexes than for the free ligands. These results could be indicative that complexation of the quinoxaline-2-carbonitrile N1,N4-dioxide derivatives with vanadium could improve their bioavailability. In addition, a new aspect of the series has been investigated. A detailed comparison of the electrochemical behavior of the free ligands and the complexes has been performed searching for a correlation between reduction potentials of the complexes and their activities and hypoxia selectivities. PMID- 16698090 TI - Dynamics of interferon-beta modulated mRNA biomarkers in multiple sclerosis patients with anti-interferon-beta neutralizing antibodies. AB - The objective of this study was to evaluate multiple interferon (IFN) specific mRNA biomarkers in multiple sclerosis (MS) patients with anti-IFN-beta neutralizing antibodies (NAB) using a pharmacodynamic study design. Thirty patients were enrolled. Blood samples were drawn at pre-treatment, 4-, 8-h time points following the intramuscular dose of IFN-beta-1a. Total RNA was obtained from peripheral blood cells, processed to cDNA and analyzed using quantitative real-time polymerase chain reaction. Pre-treatment serum samples were analyzed for anti-IFN-beta binding and neutralizing antibodies: 22 patients were NAB negative; equal numbers of the eight remaining patients were either NAB positive or had borderline NAB status. The results showed that early assessment (at 4 h after IFN-beta injection) of mRNAs for Stat-1, MxA, MxB and TRAIL was more sensitive than the later measurements. Furthermore, the NAB positive patients had strongly attenuated gene expression responses on all the mRNAs. Patients with borderline NAB had average responses that appear to be lower than NAB negative patients on several genes, notably Stat-1, TRAIL and beta2 microglobulin. PMID- 16698091 TI - Altered maturation of circulating dendritic cells in primary progressive MS patients. AB - We investigated the phenotype and frequency of circulating myeloid dendritic cells (MDC) and plasmacytoid DC (PDC) in 86 multiple sclerosis (MS) patients and 33 healthy controls (HC). The MS group comprised 20 patients with primary progressive MS (PPMS), 20 patients with secondary progressive MS (SPMS), and 46 patients with relapsing-remitting MS (RRMS) [23 treated with interferon-beta (IFN beta)]. The frequency of circulating MDC and PDC, and the expression of CD83, CD123, CD80, CD86, and CD40 were analyzed by flow cytometry. The percentage of circulating MDC was decreased in patients with SPMS and PPMS. The expression of CD83, CD80, and CD86 was lower in PPMS patients. Treatment with IFN-beta induced the expression of CD123 in PDC and decreased the number of circulating MDC. These results suggest an impaired maturation state of DC in PPMS patients, and a beneficial effect of IFN-beta favouring the survival of PDC and promoting a Th2 environment. PMID- 16698093 TI - Antiviral therapy for adenovirus infections. AB - The treatment of severe adenovirus keratoconjunctivitis and life-threatening adenovirus infections in immunocompromised patients is still unsatisfactory. We here review the mode of action and antiviral data for cidofovir and ribavirin, obtained in cell culture, animal models or patients. Several nucleoside or nucleotide analogues have been described that target the adenovirus polymerase, whereas other antiviral targets have been poorly investigated. Furthermore, optimal therapeutic response may be achieved by combining antiviral therapy with immunotherapeutic approaches, as currently being explored. PMID- 16698092 TI - Circulating dendritic cells subsets and regulatory T-cells at multiple sclerosis relapse: differential short-term changes on corticosteroids therapy. AB - Glucocorticoids remain the treatment of choice for MS relapses. However, little is known on the effect of intravenous methylprednisolone (IVMP) on dendritic cells (DCs) and regulatory T-cells (TReg). Our main goal was to quantify circulating myeloid and plasmacytoid DCs (mDCs and pDCs), and TReg at MS relapse versus healthy controls; and to analyse the short-term changes after IVMP for MS relapse. MS patients at relapse compared to controls showed higher %CD4+CD25high+ TReg (p<0.01). After 5-days of IVMP, activated T-lymphocytes (p=0.001), pDCs (p<0.0001), and CD11c+ mDCs (p<0.0001) decreased. By contrast, CD4+CD25+ and CD4+CD25high+ TReg further increased (p<0.0001 both). Changes on these subsets may play a relevant role in the immunosuppressive activity of this drug. PMID- 16698094 TI - Hippocampal evoked potentials in novel environments: a behavioral clamping method. AB - The hippocampus is involved in the detection of novelty and is essential for certain forms of learning about environmental events and relationships. The cellular and molecular mechanisms of one form of hippocampal synaptic plasticity, long-term potentiation (LTP), are thought to overlap significantly with the neural mechanisms of learning. In this study changes in hippocampal synaptic efficacy were measured in awake, freely behaving rats during exploration of novel environments. Because hippocampal physiology is modulated by on-going behavior, evoked potentials collected during Type 1 versus Type 2 behavior were evaluated separately. The effect of prior LTP induction at perforant path-dentate synapses on exploration-induced changes was evaluated. The results show that exploration causes an increase in population spike amplitude with no change in excitatory postsynaptic potential during Type 1 behavior that lasts longer than 5 min. Prior induction of hippocampal LTP occludes the change induced by exploration. This change is not likely to be due to a reduction of GABAergic inhibition induced by novelty. PMID- 16698095 TI - Post-training and post-reactivation administration of amphetamine enhances morphine conditioned place preference. AB - Amphetamine has been shown to enhance consolidation in a variety of memory paradigms. However, it is not known if amphetamine can modulate the consolidation of the types of context-reward associations involved in drug addiction, such as those formed in the conditioned place preference (CPP) task. Also, some types of memory exhibit a second period of lability following memory reactivation, and it is not known whether amphetamine administered during this period can modulate CPP. Our study investigated whether amphetamine can enhance morphine CPP when administered during the consolidation period or the post-reactivation period. Subjects were trained in the CPP task and injected with amphetamine or vehicle immediately or 6 h after each training session. The day after the completion of training, they were tested. Amphetamine injected immediately but not 6 h after training enhanced morphine CPP. In separate experiments, subjects were first trained in the CPP task. The day following the completion of training, subjects were given a memory reactivation session and injected with amphetamine or vehicle immediately or 6 h after reactivation. Subjects were tested the next day. Amphetamine injected immediately but not 6 h after memory reactivation enhanced morphine CPP. However, amphetamine injected without memory reactivation had no effect on the expression of morphine CPP. Our results suggest that amphetamine enhances the consolidation of morphine CPP and that morphine CPP exhibits a temporally limited period of post-reactivation lability during which the memory can be modulated. PMID- 16698096 TI - REMOVED: Boron neutron capture therapy (BNCT) for glioblastoma multiforme: A phase 2 study evaluating a prolonged high dose of boronophenylalanine (BPA). AB - This article has been removed consistent with Elsevier Policy on Article Withdrawal. Please see .The Publisher apologizes for any inconvenience this may cause. PMID- 16698097 TI - Anatomy changes in radiotherapy detected using portal imaging. AB - BACKGROUND AND PURPOSE: Localisation images normally acquired to verify patient positioning also contain information about the patient's internal anatomy. The aim of this study was to investigate the anatomical changes observed in localisation images and examples of dosimetric consequences. PATIENTS AND METHODS: Localisation images were obtained weekly prior to radiotherapy with an electronic portal imaging device (EPID). A series of 'difference images' was created by subtracting the first localisation image from that of subsequent fractions. Images from 81 lung, 40 head and neck and 34 prostate cancer patients were classified according to the changes observed. Changes were considered relevant if the average pixel value over an area of at least 1cm(2) differed by more than 5%, to allow for variations in linac output and EPID signal. Two patients were selected to illustrate the dosimetric effects of relevant changes. Their plans were re-calculated with repeat CT scans acquired after 4 weeks of treatment and compared with the difference images of the corresponding days. RESULTS: Progressive changes were detected for 57% of lung and 37% of head and neck cancer patients studied. Random changes were observed in 37% of lung, 28% of head and neck and 82% of prostate cancer patients. For a lung case, an increase of 10.0% in EPID dose due to tumour shrinkage corresponded to an increase of 9.8% in mean lung dose. Gas pockets in the rectum region of the prostate case increased the EPID dose by 6.3%, and resulted in a decrease of the minimum dose to the planning target volume of 26.4%. CONCLUSIONS: Difference images are an efficient means of qualitatively detecting anatomical changes for various treatment sites in clinical practice. They can be used to identify changes for a particular patient, to indicate if the dose delivered to the patient would differ from planning and to detect if there is a need for re-planning. PMID- 16698098 TI - A novel radiation therapy technique for malignant pleural mesothelioma combining electrons with intensity-modulated photons. AB - BACKGROUND AND PURPOSE: To investigate the feasibility and potential benefits of combining electron and photon intensity modulated radiotherapy (IMRT) for patients with malignant pleural mesothelioma (MPM). PATIENTS AND METHODS: The planning CT images of 11 MPM patients, six after extrapleural pneumonectomy (EPP) and five after pleurectomy/decortication (P/D), were used for this study. These cases were planned with photon IMRT alone and photon IMRT combined with electrons (IMRT+e). The latter approach incorporated the electron dose into the inverse planning optimization. The resulting doses to the planning target volume (PTV) and relevant critical structures were compared. RESULTS: For all patients, the PTV was well covered and doses to critical structures were clinically acceptable for all patients with both techniques. However, IMRT+e exhibited a distinct advantage in reducing the doses to the liver, ipsilateral kidney, contralateral kidney, and heart (P=0.002, 0.003, 0.025, and 0.001, respectively). CONCLUSIONS: This study showed that IMRT or IMRT+e is a viable treatment modality for MPM patients. Both plans can provide excellent target coverage and normal tissue sparing, but with the addition of electron beams, the critical structures can be further spared. Additional refining of the electron contribution is expected to further reduce radiation-induced morbidity. PMID- 16698099 TI - Long-term results of hypofractionated radiotherapy and hormonal therapy without surgery for breast cancer in elderly patients. AB - BACKGROUND AND PURPOSE: To evaluate early and late reactions, local control, disease-free survival, cause-specific survival, and overall survival of elderly breast cancer patients treated with definitive once-a-week hypofractionated radiotherapy together with hormonal therapy. PATIENTS AND METHODS: Between 1987 and 1999, 115 patients with a median age of 83 presenting with 124 non-metastatic breast carcinoma were treated with definitive once weekly hypofractionated radiotherapy associated with hormonal therapy. The main reasons for adopting this schedule were patient refusal of surgery, very old age, locally advanced case, and/or comorbid disease. Radiation was delivered as once-a-week, 6.5 Gy for a total breast dose of 32.5 Gy in five fractions, followed with 1-3 fractions of 6.5 Gy to the tumour site. The median follow-up was 41 months. RESULTS: Neoadjuvant hormonal therapy led to 56% reduction of the tumour volume. Late reactions occurred in 46 patients; they were mild to moderate in 87% of these patients. The Kaplan-Meier rate was 52% of patients, with 6% experiencing grade 3 reactions. The 5-year local progression-free rate was 78%. The corresponding cause-specific survival was 71%, and was influenced by T classification, nodal status, oestrogen receptors and the total dose. The first three factors retained an independent prognostic impact on multivariate analysis. The 5-year overall survival was 38%. It was affected by the T classification, lymph node involvement and the performance status (PS). Using a multivariate analysis, only T classification and PS were identified as independent factors regarding overall survival. CONCLUSIONS: Definitive hypofractionated radiotherapy allows a good local control, with acceptable toxicity. This schedule associated with hormonal therapy is a good alternative to surgery in non-operable old patients and in case of patient refusal to surgery and to standard fractionation. PMID- 16698100 TI - Does 4 MV perform better compared to 6 MV in the presence of air cavities in the head and neck region? AB - BACKGROUND AND PURPOSE: The underdose near air cavities in the head and neck region at photon energies of 4 MV and 6 MV was studied in search for clinical advantages of the 4 MV over 6 MV treatments. MATERIALS AND METHODS: The on-axis and off-axis dose distributions were measured with a parallel-plate ionization chamber and films in polystyrene phantoms containing an air cavity of appropriate size based on the results of computed tomography scans. RESULTS: Although most results are similar for both energies, the 4 MV photon beams give a somewhat smaller underdose effect and a faster re-build up than the 6 MV. For both energies a significant underdose effect was observed at the edge of the field in the larynx phantom. This proved to be true for small and large fields, for smaller and larger cavities, for one-beam as well as parallel-opposed beams. CONCLUSION: For most clinically relevant situations there is no remarkable benefit in the use of either of the two energies. PMID- 16698101 TI - Analysis of functional polymorphisms in three synaptic plasticity-related genes (BDNF, COMT AND UCHL1) in Alzheimer's disease in Colombia. AB - In recent years, it has been proposed that synaptic dysfunction may be an important etiological factor for Alzheimer's disease (AD). This hypothesis has important implications for the analysis of AD genetic risk in case-control studies. In the present work, we analyzed common functional polymorphisms in three synaptic plasticity-related genes (brain-derived neurotrophic factor, BDNF Val66Met; catechol-O-methyl transferase, COMT Val158; ubiquitin carboxyl-terminal hydroxylase, UCHL1 S18Y) in a sample of 102 AD cases and 168 age and sex matched controls living in Bogota, Colombia. There was not association between UCHL1 polymorphism and AD in our sample. We have found an initial association with BDNF polymorphism in familial cases and with COMT polymorphism in male and sporadic patients. These initial associations were lost after Bonferroni correction for multiple testing. Unadjusted results may be compatible with the expected functional effect of variations in these genes on pathological memory and cognitive dysfunction, as has been implicated in animal and cell models and also from neuropsychological analysis of normal subjects carriers of the AD associated genotypes. An exploration of functional variants in these and in other synaptic plasticity-related genes (a synaptogenomics approach) in independent larger samples will be important to discover new genes associated with AD. PMID- 16698102 TI - Black aspergilli and ochratoxin A production in French vineyards. AB - A survey on the occurrence on grape of black Aspergillus species and their capability to produce ochratoxin A (OTA) was conducted in France over three years (2001-2003) in 10 vineyards from four winemaking regions with different geographical locations and climatic conditions. During 2001 and 2002, from setting to harvest, the total numbers of fungal isolates were respectively 721 and 711 increasing in 2003 to reach 1035. The Aspergillus genus was essentially represented by Section Nigri (99%) and it was predominant (80%+/-4.6) when compared to Penicillium (20%+/-4.6). Regardless of sampling year, 32.5% (+/ sigma=1.26) of the fungal isolates were OTA producers and 93% (+/-sigma=2.65) belonging to black aspergilli. The ochratoxigenic potential of the isolates and their occurrence on grapes revealed that Aspergillus carbonarius was the main OTA producer (up to 37.5 mug/g). At harvest time, the fungal population was maximal and this was the most critical period influencing OTA contamination. Grapes from Languedoc-Roussillon region were most infested with ochratoxigenic fungi and had the highest concentrations of OTA (up to 2.8 ng/g). PMID- 16698103 TI - Occurrence of ochratoxin A producing fungi in wine and table grapes in Israel. AB - A 3-year survey was conducted to assay the number of Aspergillus Section Nigri isolates and in vitro ochratoxin A (OTA) production capacity in 10 vineyards in Israel. The survey included field sampling of two wine cultivars, 'Sauvignon Blanc' and 'Cabernet Sauvignon' as well as the table grape cultivar 'Superior'. A total of 2114 isolates were analyzed and of those 161 isolates were shown to produce OTA. The major finding was that Aspergillus carbonarius (336 tested strains) is the most consistent producer of OTA, with approximately 35% of the isolates identified as positive in vitro. In comparison, 3.1% of other isolates from the Aspergillus niger aggregate (of 1432 strains) produced OTA in vitro. In contrast, none of the 346 tested strains with a uniseriate head morphology produced OTA. The incidence of infected berries was very low before veraison, while at harvest, this frequency was twice as high. In general, the composition of black Aspergilli did not differ during berry development. Generally, more OTA producing isolates were isolated from the surface of table grapes cv. 'Superior' compared to 'Sauvignon Blanc'. None of the samples collected at harvest contained traces of OTA in the juice. This study shows that grapes in Israel are contaminated with ochratoxigenic species which represent a risk of OTA contamination. PMID- 16698104 TI - Application of global analysis techniques to Corynebacterium glutamicum: new insights into nitrogen regulation. AB - The regulation of nitrogen metabolism in the amino acid producer Corynebacterium glutamicum was subject of research for several decades. While previous studies focused on single enzymes or pathways, the publication of the C. glutamicum genome sequence gave a fresh impetus to research, since a global investigation of metabolism and regulation networks became possible based on these data. This communication summarizes the advances made by different studies, in which global analysis approaches were used to characterize the C. glutamicum nitrogen starvation response. A combination of bioinformatics approaches, transcriptome and proteome analyses as well as chemostat experiments revealed new insights into the nitrogen control network of C. glutamicum. C. glutamicum reacts to a limited nitrogen supply with a rearrangement of the cellular transport capacity, changes in metabolic pathways for nitrogen assimilation and amino acid biosynthesis, an increased energy generation and increased protein stability. With the aid of chemostat experiments, in which different growth rates were obtained by nitrogen limitation, general starvation effects could be distinguished from specific nitrogen limitation-dependent changes. The core adaptations on the level of transcription are controlled by the master regulator of nitrogen control, the TetR-type protein AmtR. This global regulator governs transcription of at least 33 genes via binding to a palindromic consensus motif (AmtR box). Genes with AmtR box-containing promoters were identified by genome-wide screening and validated, besides by other methods, by transcriptome analyses using DNA microarrays. PMID- 16698105 TI - Antibody-based metabolic engineering in plants. AB - Genetic engineering is a powerful tool for the manipulation of cellular metabolism and the development of plant varieties with enhanced biological and nutrional functions. Several strategies are available for the in vivo modulation of enzymatic activities, allowing metabolic flux to be directed towards desired biochemical products. Such strategies include the simultaneous expression and/or suppression of multiple genes encoding rate-limiting enzymes, ectopic expression of transcription factors, and the RNA-based inhibition of catabolic enzymes. As an alternative approach, recombinant antibodies expressed in plants have been used to inactivate or sequestrate specific host proteins or compounds, resulting in significant changes to metabolic pathways. The impact of this approach depends on prudent selection of the target antigen, careful antibody design, appropriate subcellular targeting and stable accumulation of the recombinant antibodies in planta. Here, we describe the current status of antibody-based metabolic engineering in plants, discuss procedures for the optimisation of this technology and consider the remaining challenges to its widespread use. PMID- 16698106 TI - High efficient degradation of dyes with lignin peroxidase coupled with glucose oxidase. AB - The H(2)O(2) supply strategy was one of crucial factors for high efficient degradation of pollutants with lignin peroxidase (LiP). In this paper, an attempt was made to couple a H(2)O(2) producing enzymatic reaction to the LiP catalyzed oxidation of dyes. H(2)O(2) needed was generated by glucose oxidase (GOD) and its substrate glucose. The generation rate of H(2)O(2) could be easily controlled by adjusting the pH of the degradation system and the amount of GOD added. Due to the controlled release of H(2)O(2), a sustainable constant activity of LiP was observed. The inhibition of LiP by high level H(2)O(2) supplied externally by a single addition at the beginning of the experiments could be avoided. Degradation of three dyes (xylene cyanol, fuchsine and rhodamine B) with LiP coupled with GOD indicated that the present H(2)O(2) supply strategy was very effective for improvement of the efficiency of the decolourization of dyes. PMID- 16698107 TI - Saccharomyces cerevisiae: a useful model host to study fundamental biology of viral replication. AB - Understanding the fundamental steps of virus life cycles including virus-host interactions is essential for the design of effective antiviral strategies. Such understanding has been deferred by the complexity of higher eukaryotic host organisms. To circumvent experimental difficulties associated with this, systems were developed to replicate viruses in the yeast Saccharomyces cerevisiae. The systems include viruses with RNA and DNA genomes that infect plants, animals and humans. By using the powerful methodologies available for yeast genetic analysis, fundamental processes occurring during virus replication have been brought to light. Here, we review the different viruses able to direct replication and gene expression in yeast and discuss their main contributions in the understanding of virus biology. PMID- 16698108 TI - Chronic fatigue syndrome: a systematic review. AB - OBJECTIVE: A systematic review of the literature about chronic fatigue syndrome (CFS). METHODS: A search of the Medline database (via Ovid and PubMed) with the key words chronic fatigue syndrome, diagnosis, classification, epidemiology, etiology, physiopathology, metabolism, microbiology, immunology, virology, psychology, drug therapy, rehabilitation, and therapy. The reference lists of each article were examined for additional related articles. RESULTS: CFS was defined in 1988 by the US Centes for Disease Control and Prevention. The prevalence of chronic fatigue syndrome has ranged from 0.2% to 0.7% in the general population. In 1994, the definition of CFS was revised by Fukuda et al. Despite various research in several topics (e.g. infection, immune systems, neuroendocrinology, autonomic activity, neuromuscular involvement), the pathophysiology remains unknown. CONCLUSION: CFS, with its various major clinical and functional impacts, should be associated with a "biopsychosocial model". Progressive muscular rehabilitation, combined with behavioral and cognitive treatment, is an essential part of therapy. PMID- 16698109 TI - Perineal neuromuscular fatigue. AB - AIM: The physiology of urinary continence during stress is complex and the role of passive and active mechanisms remains unclear. Coughing leads to a contraction of urethral rhabdomyosphincter and pelvic floor muscles leading to a positive urethro-vesical gradient and continence. Neuromuscular fatigue can involve all striated muscles, including rhabdomyosphincter, peri-urethral and pelvic floor muscles. This article reviews results of studies assessing perineal muscular fatigue in urinary incontinence. MATERIALS AND METHODS: A systematic review of the literature (Medline, Pascal and Embase) with use of the MESH keywords fatigue, stress, urinary incontinence, pelvic floor, urethra, urethral pressure, and muscle. RESULTS: Animal models have shown that the pelvic muscles (iliococcygeus and pubococcygeous) exhibit more neuromuscular fatigue than classical skeletal striated muscles (i.e. soleus muscle). Although the human external urethral sphincter is considered to be a highly fatigue-resistant muscle with its high proportion of slow muscle fibers, repeated coughing seems to lead to decreased urethral pressure in numerous women affected with stress urinary incontinence. In this case, "urethral fatigue" might be a possibility. CONCLUSIONS: Although few studies have focused on perineal muscular fatigue, such increased fatigue in pelvic floor muscles may play a role in the pathophysiologic features of stress urinary incontinence in women. PMID- 16698110 TI - [Intrathecal baclofen. Twenty years later]. PMID- 16698111 TI - Portal vein ligation and partial hepatectomy differentially influence growth of intrahepatic metastasis and liver regeneration in mice. AB - BACKGROUND/AIMS: Since modern treatment of liver metastases includes serial hepatectomies and portal vein obstruction, we investigated the effects of portal vein ligation (PVL) and partial hepatectomy (PH) on tumor growth. METHODS: The effects of 70% PH and PVL on liver and lung metastases were evaluated in mice. Tumor growth and liver regeneration were assessed by morphometry and immunohistochemistry for PCNA and BrdU. The effect of growth factors of liver regeneration on CT-26 cells was tested in vitro, and TGF-beta secretion of CT-26 cells was measured by ELISA. RNA synthesis of TGF-beta and activin A was measured by RT-PCR. RESULTS: Liver regeneration after PH and PVL was similar in tumor-free mice. Intrahepatic tumor growth was lower after PH than after PVL (p=0.016). Extrahepatic tumor growth was not different. In contrast to PVL, liver regeneration was delayed after PH in metastatic livers (p=0.001). Tested growth factors of liver regeneration stimulated CT-26 cells in vitro, and CT-26 secreted significant amounts of TGF-beta in vitro and in vivo. CONCLUSIONS: Although similar in tumor-free mice, liver regeneration significantly differed between PVL and PH in metastatic livers. In addition, PH and PVL differently affected intrahepatic tumor growth. PMID- 16698112 TI - Limiting youth access to tobacco: comparing the long-term health impacts of increasing cigarette excise taxes and raising the legal smoking age to 21 in the United States. AB - Although many states in the US have raised cigarette excise taxes in recent years, the size of these increases have been fairly modest (resulting in a 15% increase in the per pack purchase price), and their impact on adult smoking prevalence is likely insufficient to meet Healthy People 2010 objectives. This paper presents the results of a 75-year dynamic simulation model comparing the long-term health benefits to society of various levels of tax increase to a viable alternative: limiting youth access to cigarettes by raising the legal purchase age to 21. If youth smoking initiation is delayed as assumed in the model, increasing the smoking age would have a minimal immediate effect on adult smoking prevalence and population health, but would affect a large drop in youth smoking prevalence from 22% to under 9% for the 15-17-year-old age group in 7 years (by 2010)-better than the result of raising taxes to increase the purchase price of cigarettes by 100%. Reducing youth initiation by enforcing a higher smoking age would reduce adult smoking prevalence in the long-term (75 years in the future) to 13.6% (comparable to a 40% tax-induced price increase), and would produce a cumulative gain of 109 million QALYs (comparable to a 20% price increase). If the political climate continues to favor only moderate cigarette excise tax increases, raising the smoking age should be considered to reduce the health burden of smoking on society. The health benefits of large tax increases, however, would be greater and would accrue faster than raising the minimum legal purchase age for cigarettes. PMID- 16698113 TI - Life saving or money wasting? Perceptions of caesarean sections among users of services in rural Bangladesh. AB - Bangladesh has a high level of maternal mortality, corresponding to one of the world's lowest rates of use of skilled birth attendance (12.1%), and a similarly low rate of caesarean births (2.4%). While increasing the proportion of women who deliver with professional medical care is essential to prevent maternal deaths, past work has identified distrust of caesarean procedures in Bangladesh. The reasons behind this distrust can manifest itself in health seeking behaviour around maternal care. This paper presents findings from a qualitative study of 30 women in a rural district of Bangladesh who recently delivered in a health facility. It finds that the distrust in doctor's recommendations for surgery stemmed from high costs incurred and a belief that it was used when not medically justified. This could lead to women avoiding or leaving medical facilities in extreme cases. Some women's experiences further illustrated disagreement among medical staff as to whether or not a caesarean procedure should be done, with conflicting financial incentives for doctors to perform caesarean deliveries, and for nurses and midwives to conduct normal deliveries. Policy makers must recognise that the fears women hold of caesarean deliveries may not simply be rooted in ignorance and may, in fact, reflect legitimate concerns with medical practice. Ultimately, it will be essential to address problems in the health systems environment, which may promote improper service provision. PMID- 16698114 TI - Plasma fibrinogen and serum C-reactive protein are associated with non-small cell lung cancer. AB - OBJECTIVES: There is an association between coagulation and lung cancer. Therefore, pre-operative plasma fibrinogen and serum C-reactive protein (CRP) concentration were assessed to determine their association with tumour characteristics and to ascertain any role in patient selection for curative resection. METHODS: These parameters were compared with tumour size, pTNM stage, and possibility of complete resection in 93 patients with non-small cell lung cancer who underwent surgical resection. RESULTS: Plasma fibrinogen concentration (r(s)=0.34, P=0.001) and serum CRP concentration (r(s)=0.34, P=0.001) were positively correlated with maximum pathological tumour size. A higher plasma fibrinogen concentration was associated with squamous cell carcinoma versus adenocarcinoma (4.5+/-0.13 g/L versus 3.6+/-0.28 g/L; P=0.008), with a trend towards a similar association for CRP (P=0.06). Pathological T stage was also associated with mean plasma fibrinogen and serum CRP concentration (P=0.01 and 0.04, respectively), but pN stage was not associated with either parameter. Incomplete resection occurred in 23% of patients with plasma fibrinogen > 5 g/L or serum CRP > 40 mg/L (versus only 8% when fibrinogen < or = 5 g/L and CRP < or = 40 mg/L; P=0.09). CONCLUSIONS: Plasma fibrinogen and serum CRP are associated with tumour characteristics. High values were associated with inability to achieve complete resection which may refine patient selection for thoracotomy when used with other staging modalities. Attempted resection may be justified in a patient of borderline fitness who has favourable plasma fibrinogen and serum CRP concentration, where a high resection rate is possible. As the relationship was with T stage rather than N stage it may be complimentary to PET scanning, which has only marginally better accuracy for T stage than CT scanning. PMID- 16698115 TI - Evaluating mobility for radiotherapy planning of lung tumors: a comparison of virtual fluoroscopy and 4DCT. AB - PURPOSE: Fluoroscopy is widely used for evaluating tumor mobility in radiotherapy planning. Lung tumor mobility was scored using virtual fluoroscopy, and this was compared to mobility derived from contoured tumors in all phases of a respiration correlated (or 4D) CT scan. METHODS AND MATERIALS: 4DCT datasets were reviewed and 29 patients were identified in whom tumors were visible on anterior-posterior fluoroscopy views. Mobility in all directions was estimated on fluoroscopy movie loops by four clinicians. These results were compared to mobility measured from contoured tumor volumes in all phases of the same 4DCT. Internal target volumes (ITV) were generated for both approaches. RESULTS: In eight patients, fluoroscopy did not allow for tumor mobility to be assessed in at least one direction. No significant inter-clinician variation was observed with respect to fluoroscopic assessment of mobility. Clinicians systematically overestimated mobility in all three directions (p<0.05). The mean ITVs derived using fluoroscopy were 52.2% larger than those derived using 4DCT contours, but the individual ITVs were smaller in three patients. CONCLUSION: Use of virtual fluoroscopy generally overestimates the mobility of visible lung tumors, and results in irradiation of unnecessarily large target volumes. In contrast, use of 4DCT minimizes the risk of normal tissue toxicity. PMID- 16698116 TI - Time scarcity and food choices: an overview. AB - Time scarcity, the feeling of not having enough time, has been implicated in changes in food consumption patterns such as a decrease in food preparation at home, an increase in the consumption of fast foods, a decrease in family meals, and an increase in the consumption of convenience or ready-prepared foods. These food choices are associated with less healthful diets and may contribute to obesity and chronic health problems such as cardiovascular disease, diabetes, and cancer. In spite of the potential importance for health, there has been little study of how time scarcity influences people's food choices. This paper presents an overview of time issues related to food choices and discuss applications of time research for nutrition and health researchers, policy makers, and practitioners interested in food choice. PMID- 16698117 TI - Duration of operation as a risk factor for surgical site infection: comparison of English and US data. AB - T times are used to categorize surgical procedures into long and short durations. They constitute a part of the US National Nosocomial Infection Surveillance (NNIS) risk index that is widely used internationally in surveillance for surgical site infections (SSIs). The objective of this study was to compare the US NNIS T times with data collected in England. The Surgical Site Infection Surveillance Service in England holds data collected by 168 hospitals in 13 categories of surgical procedures between 1997 and 2002. The 75(th) percentile and corresponding T time were calculated from English data and compared with US times. Differences in rates of SSI above and below the T times were compared. Graphical methods were used to assess the cut points that exhibited an association with risk of SSI. The results show that English and US T times were the same for all surgical categories except coronary artery bypass graft and vascular surgery, where the English T time was 4 h. The 75(th) percentile time for hip hemiarthroplasties was 40 min less than for total hip replacements (THR). Although the incidence of SSI in THR was significantly higher in operations lasting for longer than the T time (P<0.05), no association between risk of SSI and T times set at 1, 1.5 or 2 h was observed for hip hemiarthroplasties. In conclusion, operations lasting for longer than the T time were associated with a higher risk of SSI in most categories. In the hip prosthesis category, this association only applied to THR. PMID- 16698118 TI - Risk of conjunctival contamination in total joint arthroplasty. AB - This study arose from a concern about the transmission of infection through conjunctival contamination during total joint arthroplasty surgery. A prospective analysis of 110 sets of personal face and eye protective equipment (Sterishields) used for 29 total hip arthroplasties (THA) and 26 total knee arthroplasties (TKA) were examined for macroscopic contamination. In additition, a postal questionnaire regarding the perceived risk of eye contamination and current practice of eye protection used during TKA and THA surgery was sent to 1500 Fellows of the British Orthopaedic Association. All Sterishields demonstrated macroscopic contamination with an average of 203 blood and fat spots. The number of these was significantly higher in the lower half of the Sterishields. Of the 834 useable replies, 48% of respondents did not use adequate eye protection routinely. Twenty-eight percent felt that no eye protection was needed, while 45% felt that the eye protection available compromised their vision and so did not use it. One hundred and seven respondents (13%) reported being aware of eye contamination, while 21 (2.5%) respondents were aware of at least one case of conjunctival infection following contamination. This study demonstrates that there is a high risk of conjunctival contamination with fat and blood in major total joint arthroplasty. The reported use of appropriate eye and face protection in the UK has been shown to be low (52%). PMID- 16698119 TI - In vitro antimicrobial activity of the novel polymeric guanidine Akacid plus. AB - The bactericidal and fungicidal activity of Akacid plus, a novel polymeric compound of the cationic family of disinfectants, was compared with chlorhexidine digluconate using quality control strains of Staphylococcus aureus, Enterococcus hirae, Escherichia coli, Pseudomonas aeruginosa, Candida albicans and Aspergillus niger. In vitro activity was determined using the quantitative suspension tests described by the European Committee for Standardization. These use concentrations of 0.01-0.5% against bacterial strains/C. albicans, with 0.5-4% against A. niger, and exposure times of 5, 15 and 60 min in the presence and absence of 0.3% bovine albumin and with dilution in distilled and hard water. In the basic quantitative suspension test, Akacid plus destroyed all bacterial pathogens at a concentration of 0.1% in < or =5 min. Chlorhexidine was also highly active against S. aureus, E. coli and P. aeruginosa, but failed to eliminate E. hirae within 5 min. A high organic load reduced the bactericidal activity of both disinfectants slightly. Akacid plus showed fungicidal activity against C. albicans within 15-60 min and eliminated A. niger at a concentration of 1% in 5 min of contact. Chlorhexidine was fungicidal against C. albicans, but not against A. niger. PMID- 16698120 TI - Laboratory and patient record audit of ESBL-positive enterobacteriaceae. PMID- 16698121 TI - Dopamine turnover is upregulated in the caudate/putamen of asymptomatic MPTP treated rhesus monkeys. AB - In Parkinson's disease (PD) and experimental parkinsonism, losses of up to 60% and 80%, respectively, of dopaminergic neurons in substantia nigra, and dopamine (DA) in striatum remain asymptomatic. Several mechanisms have been suggested for this functional compensation, the DA-mediated being the most established one. Since this mechanism was recently challenged by striatal DA analysis in 1-methyl 4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated monkeys, we present data on several DAergic parameters in three groups of rhesus monkeys: MPTP-treated asymptomatic animals; symptomatic MPTP-treated animals with stable parkinsonism; and untreated sex and age matched controls. We determined ratios of striatal and nigral 3,4-dihydroxyphenyl acetic acid (DOPAC) to DA levels and tyrosine hydroxylase (TH) enzyme activity to DA levels, in addition to the commonly used homovanillic acid (HVA)/DA ratios which, as such, might be less reliable under the conditions of partial denervation. We found that in the asymptomatic MPTP monkeys the DOPAC/DA ratios in putamen and caudate nucleus were shifted with high statistical significance 1.9-5.8-fold, as compared to controls, the shifting of the ratios being in the same range as the 2.6-5.4-fold shifts in the symptomatic animals. Also TH/DA ratios were significantly increased in both, the asymptomatic and the symptomatic MPTP-treated monkeys, with shifts in the putamen and caudate nucleus of 3- and 2.7-7.0-fold, respectively. In the substantia nigra, DOPAC levels and TH activity were strongly decreased after MPTP (-77 to -97%), but the ratios DOPAC/DA and TH/DA were not changed in this brain region. Collectively, our findings support the concept of DAergic compensation of the progressive striatal DA loss in the presymptomatic stages of the parkinsonian disease process. PMID- 16698122 TI - Density of Sgt1-immunopositive neurons is decreased in the cerebral cortex of Alzheimer's disease brain. AB - Sgt1 was discovered as a protein required for the mitotic activity of kinetochore and for the activity of ubiquitin ligase in yeast [Kitagawa, K., Skowyra, D., Elledge, S.J., Harper, J.W., Hieter, P., 1999. SGT1 encodes an essential component of the yeast kinetochore assembly pathway and a novel subunit of the SCF ubiquitin ligase complex. Mol. Cell 4, 21-33.]. Later, Sgt1 was identified in different organisms including mammals where it was found at high level in the brain. To understand Sgt1 function in this tissue we analyzed its localization in human brain by immunohistochemistry. In normal brain we observed Sgt1 immunostaining in Purkinje cells of the cerebellum, in granule cells of the dentate gyrus of the hippocampus and in multiple neurons of the cortex. By Western blotting we found a higher level of this protein in the cortex than in the cerebellum. Subsequent morphometric analyses showed that the density of Sgt1 immunopositive neurons varied in different cortical regions. The highest density of Sgt1-immunopositive cells was seen in the temporal cortex (from 1.2% to 5.7%), and the lowest - in the entorhinal cortex (from 0 to 1.1% of all neurons). We next compared the density of Sgt1-immunopositive neurons in cortical layers of healthy aged and Alzheimer's disease (AD) brain sections. A significant decrease in Sgt1-immunopositive neurons was found in the temporal (up to 25-fold), angular (up to 11-fold) and posterior cingulate cortex (up to five-fold). In the entorhinal and precentral cortex the reduction of Sgt1-immunopositive neurons was only about two-fold in AD brains as compared to healthy aged ones. The presence of Sgt1 in post-mitotic neurons indicates the involvement of this protein in a process different from that required for activity of the kinetochore. Decreased immunostaining in AD cortex point to Sgt1 as a possible marker of neurons degenerating in AD. PMID- 16698123 TI - Evaluation of neuroendocrine status in longevity. AB - It is well known that physiological changes in the neuroendocrine system may be related to the process of aging. To assess neuroendocrine status in aging humans we studied a group of 155 women including 78 extremely old women (centenarians) aged 100-115 years, 21 early elderly women aged 64-67 years, 21 postmenopausal women aged 50-60 years and 35 younger women aged 20-50 years. Plasma NPY, leptin, glucose, insulin and lipid profiles were evaluated, and serum concentrations of pituitary, adrenal and thyroid hormones were measured. Our data revealed several differences in the neuroendocrine and metabolic status of centenarians, compared with other age groups, including the lowest serum concentrations of leptin, insulin and T3, and the highest values for prolactin. We failed to find any significant differences in TSH and cortisol levels. On the other hand, LH and FSH levels were comparable with those in the elderly and postmenopausal groups, but they were significantly higher than in younger subjects. GH concentrations in centenarians were lower than in younger women. NPY values were highest in the elderly group and lowest in young subjects. We conclude that the neuroendocrine status in centenarians is markedly different from that found in early elderly or young women. PMID- 16698124 TI - Transthyretin and Alzheimer's disease: where in the brain? AB - Transthyretin (TTR), a carrier protein for thyroxine and retinol in plasma and cerebrospinal fluid (CSF), has been shown to bind the amyloid beta peptide. Accordingly, TTR has been suggested to protect against amyloid beta deposition, a key pathological feature in Alzheimer's disease (AD). Supporting this view are the reduced TTR levels found in CSF of patients with AD, as well as reports of altered TTR expression in the cortex and hippocampus of AD rodent models. Importantly, early characterization of TTR distribution revealed the choroid plexus as the site of TTR synthesis within the brain. To resolve this controversy we used precise laser microdissection technology to assay for TTR mRNA expression. Our results clearly demonstrate that TTR is not produced in the brain parenchyma of wild-type mice nor in two different transgenic mouse models of AD, suggesting that contamination by choroid plexus contributed to the recent results indicating TTR production in various brain regions. The relevance of TTR to AD should now take into consideration TTR production by the choroid plexus and its ability, in the CSF, to sequester the amyloid beta peptide. PMID- 16698125 TI - Long-term measures of free testosterone predict regional cerebral blood flow patterns in elderly men. AB - We previously reported that high circulating free testosterone (T) was associated with better performance on tests of memory, executive function, and spatial ability, and with a reduced risk for Alzheimer's disease. In this study, we report that free T levels, measured on multiple occasions over 14 years, predict regional cerebral blood flow (rCBF) measured by PET in 40 older men. Voxel-based regression, indicated that higher Free T was associated with increased rCBF in the hippocampus bilaterally (extending to the parahippocampal gyrus on the right), anterior cingulate gyrus, and right inferior frontal cortex. Total T concentrations were positively correlated with rCBF in the left putamen, bilateral thalamus, and left inferior frontal cortex and negatively correlated with amygdala rCBF bilaterally. These findings suggest that endogenous T influences brain physiology in regions critical for memory and attention and provide one mechanism through which T may affect cognitive function. PMID- 16698126 TI - Cerebrovascular risk factors do not contribute to genetic variance of cognitive function: the ERF study. AB - Impaired cognition in later life may result from Alzheimer's disease-related pathology, but also from vascular pathology. We studied to what extent vascular risk explained heritability of cognition in 780 individuals, related in one extended pedigree in a genetically isolated population, in the ERF study. Heritability was estimated using variance components modelling (SOLAR). Univariate analyses included models with and without vascular disease; bivariate analyses included both cognitive and vascular traits, such as blood pressure, serum glucose or lipids. Heritability for immediate and delayed recall, recognition, semantic fluency, Trail making B and Stroop tests was significant, with estimates from 0.16 to 0.36. Vascular factors did not affect cognitive functions, except immediate recall and the Stroop test. Heritability estimates did not change significantly when adjusted for vascular disease. We found no genetic correlation between cognition and vascular traits. Therefore, in this population vascular disease is mildly associated with cognitive dysfunction, and in those with vascular disease, the underlying genetic risk factors are not likely to account for the genetic variation in cognition at adult age. PMID- 16698127 TI - Genetic and environmental contribution to postural balance of older women in single and dual task situations. AB - The purpose of the present study was to examine the effect of a second task on postural balance and to determine the role of genetic influences on postural balance when dual tasking among 206 monozygotic and 227 dizygotic female twins, aged 63-76 years. Balance was measured as medio-lateral and antero-posterior velocity of the centre of pressure (COP) (mm/s) and velocity moment (mm(2)/s) while standing on a force platform. Doing an arithmetic task increased movement of the COP while the hand motor task had no effect on movement of the COP. The genetic contribution to balance in the single task situation was minor (14%, 95% confidence interval, CI: 11-35%) whereas in the dual task situation it was moderate (28%, 95% CI: 0.02-42%). The finding can be explained in the light of the underlying phenotypes of dual tasking, such as cognitive processing that is found to be under considerable genetically controlled even in old age. Moreover, the present study supports previous studies suggesting that, especially among older people, when simultaneously maintaining balance and performing another cognitively demanding task, additional central processing is recruited. PMID- 16698128 TI - Plasma beta amyloid and impaired CO2-induced cerebral vasomotor reactivity. AB - Amyloid beta (Abeta) may disturb cerebral autoregulation by damaging the wall of small cerebral blood vessels and by direct negative vasoactive properties. We assessed whether previous and concurrent plasma Abeta(1-40) and Abeta(1-42) levels were associated with an impaired CO2-induced cerebral vasomotor response. In the longitudinal population-based Rotterdam Study we measured plasma Abeta levels and cerebral vasomotor reactivity to hypercapnia with transcranial Doppler ultrasonography (TCD) in 441 people, aged 60-90 years. We performed age and sex adjusted logistic regression analysis. Plasma Abeta levels assessed on average 6.5-year before TCD were linearly associated with an impaired CO2-induced cerebral vasomotor response (odds ratio 1.48 (95%CI 1.19;1.84) per standard deviation increase in Abeta(1-40), and 1.36 (95%CI 1.09;1.70) per standard deviation increase in Abeta(1-42)). Such an association was not present for Abeta assessed concurrently with the TCD measurement. Persons whose plasma Abeta(1-40) levels had decreased in the 6.5-year period preceding TCD measurements were more likely to have an impaired CO2-induced vasomotor reactivity. Overall our observations are most compatible with plasma Abeta levels representing vascular Abeta deposits years later resulting in impaired CO2-induced vasomotor reactivity. PMID- 16698129 TI - ESPEN Guidelines on Enteral Nutrition: Gastroenterology. AB - Undernutrition as well as specific nutrient deficiencies have been described in patients with Crohn's disease (CD), ulcerative colitis (UC) and short bowel syndrome (SBS). The present guideline gives evidence-based recommendations for the indication, application and type of formula of enteral nutrition (EN) (oral nutritional supplements (ONS) or tube feeding (TF)) in these patients. It was developed in an interdisciplinary consensus-based process in accordance with officially accepted standards and is based on all relevant publications since 1985. ONS and/or TF in addition to normal food is indicated in undernourished patients with CD or CU to improve nutritional status. In active CD EN is the first line therapy in children and should be used as sole therapy in adults mainly when treatment with corticosteroids is not feasible. No significant differences have been shown in the effects of free amino acid, peptide-based and whole protein formulae for TF. In remission ONS is recommended only in steroid dependent patients in CD. In patients with SBS TF should be introduced in the adaptation phase and should be changed with progressing adaptation to ONS in addition to normal food. PMID- 16698130 TI - A prospective study on weight loss and energy intake in patients with head and neck cancer, during diagnosis, treatment and revalidation. AB - BACKGROUND & AIMS: Patients with head and neck carcinomas often loose a significant percentage of weight, which correlates with the complication rate. Only limited information is available on the moment and extent of weight loss and energy intake in time and the relation with type of treatment. The aim of this study was to describe the moment and extent of weight loss and nutritional energy intake in patients with tumors in oral cavity, oropharynx and hypopharynx during diagnosis, treatment and revalidation. METHODS: An observational, prospective study on weight changes and nutritional intake in these patients was carried out during diagnosis, treatment and revalidation in relation to the type of treatment. RESULTS: Forty-seven patients successfully completed the study. A significant difference in mean bodyweight was found for patients treated by radiotherapy during treatment (-3.3 kg, P=0.01) and the early revalidation period (-3.4 kg, P=0.01) and for patients treated with surgery during diagnoses (-1.5 kg, P=0.001) and early revalidation period (1.6 kg, P=0.02). Overall patients lowered their energy intake by 122 kcal/day followed by a significant increase in energy intake during revalidation 326 kcal/day (P=0.04). CONCLUSIONS: The radiotherapy and the concomitant radio-chemotherapy group lost most body weight during treatment and early revalidation. All treatment groups experienced a decrease in energy intake during treatment followed by a significant increase during revalidation. PMID- 16698131 TI - Tolerable infusion rate of citrate based on clinical signs and the electrocardiogram in conscious dogs. AB - BACKGROUND & AIMS: The possible clinical significance of the toxic effects of citrate has not yet been fully clarified. This study was therefore conducted to confirm the toxicity and determine the tolerable infusion rate of citrate administered by rapid intravenous infusion to conscious dogs. METHODS: Citrate solutions were infused via the cephalic vein of 4 conscious dogs at 0.33, 0.67, or 1.33mmol/kg/h up to 1.33mmol/kg. Clinical signs and the electrocardiogram were observed during and after infusion. Serum citrate and ionized calcium levels were also measured. RESULTS: Although the mean citrate level increased in accordance with the infusion rate, the calcium level decreased. No significant changes in clinical signs or the electrocardiogram were observed during infusion at 0.33mmol/kg/h despite an increase in the serum citrate level to 1.22+/-0.11mmol/l (pre-infusion value: 0.38+/-0.01mmol/l) and a decrease in the serum calcium level to 1.28+/-0.03mmol/l (pre-infusion value: 1.50+/-0.05mmol/l). Vomiting and QTc prolongation were observed at 0.67mmol/kg/h or higher. Salivation and tachycardia were observed at 1.33mmol/kg/h. CONCLUSIONS: Based on clinical signs and the electrocardiogram, the tolerable infusion rate of citrate in conscious dogs is concluded to be 0.33mmol/kg/h. PMID- 16698132 TI - The German hospital malnutrition study. AB - BACKGROUND & AIMS: Malnutrition is frequently observed in chronic and severe diseases and associated with impaired outcome. In Germany general data on prevalence and impact of hospital malnutrition are missing. METHODS: Nutritional state was assessed by subjective global assessment (SGA) and by anthropometric measurements in 1,886 consecutively admitted patients in 13 hospitals (n=1,073, university hospitals; n=813, community or teaching hospitals). Risk factors for malnutrition and the impact of nutritional status on length of hospital stay were analyzed. RESULTS: Malnutrition was diagnosed in 27.4% of patients according to SGA. A low arm muscle area and arm fat area were observed in 11.3% and 17.1%, respectively. Forty-three % of patients 70 years old were malnourished compared to only 7.8% of patients <30 years. The highest prevalence of malnutrition was observed in geriatric (56.2%), oncology (37.6%), and gastroenterology (32.6%) departments. Multivariate analysis revealed three independent risk factors: higher age, polypharmacy, and malignant disease (all P<0.01). Malnutrition was associated with an 43% increase of hospital stay (P<0.001). CONCLUSIONS: In German hospitals every fourth patient is malnourished. Malnutrition is associated with increased length of hospital stay. Higher age, malignant disease and major comorbidity were found to be the main contributors to malnutrition. Adequate nutritional support should be initiated in order to optimize the clinical outcome of these patients. PMID- 16698133 TI - Pulmonary function and sleep-related breathing disorders in severely obese children. AB - BACKGROUND & AIMS: To evaluate the frequency of pulmonary function and sleep breathing disorders in severely obese children and to search for their association with obesity phenotypes. METHODS: Sleep studies and spirometry were performed for 54 severely obese children. RESULTS: Upper airway resistances (RAWs) were increased with RAW>200% and forced 25s expiratory volume<80% in 83% and 60% of individuals, respectively. A decrease in functional residual capacity (FRC)<80% was found in 43%. Fifty-two percent of the children had a desaturation index>10 during sleep, and 41% of children presented at least one of three severity criteria (snoring index>300 per hour, respiratory events index (REI)>10 and arousal index>10). Univariate analyses showed a positive correlation between snoring index and BMI Z-score and neck/height ratio (P=0.01 and 0.04, respectively) as between REI and the same parameters (P=0.01 and 0.03, respectively). In a multivariate model with BMI Z-score, NHR still correlated with the snoring index (P=0.02) and REI (P=0.01). CONCLUSIONS: In our cohort, obese children showed frequent pulmonary function and sleep-breathing disorders. The later were associated with impaired upper airway respiratory conductance. PMID- 16698134 TI - The effect of the ingestion of Ginkgo biloba extract (EGb 761) on the pharmacokinetics of metformin in non-diabetic and type 2 diabetic subjects--a double blind placebo-controlled, crossover study. AB - BACKGROUND & AIMS: Ginkgo biloba extract (EGb 761) has been shown to ameliorate some defects associated with the insulin resistance syndrome and so patients with Type 2 diabetes mellitus (T2DM) may be inclined to co-ingest the herb with their medications, such as metformin. This study was designed to determine if the co ingestion of EGb 761 and metformin would alter the pharmacokinetic properties of metformin in T2DM patients and persons without diabetes, who may ingest it for other purposes. METHOD: Normal glucose tolerance (NGT) subjects (n=10; age, 39.2+/-14.0 years; fasting plasma glucose (FPG), 90+/-7 mg/dl; body mass index (BMI), 24.1+/-3.7 kg/m(2)) and 10 T2DM patients (n=10; age, 51.7+/-8.9 years; FPG, 150+/-7 mg/dl; BMI, 33.7+/-5.7 kg/m(2)) completed a randomized, double blind, placebo-controlled crossover study. They ingested either EGb 761 (12 0mg/day as a single dose) or a vegetable-based placebo during each arm for 3 months. At the end of each arm, the NGT subject ingested a single 500 mg dose of metformin (non-diabetics) and the T2DM subject took his/her prescribed metformin dose (250-850 mg) with 120 mg EGb 761. Blood and urine samples were collected over an 8-h period, and in the case of T2DM subjects, additionally over the first 2h of the subsequent 3 days. RESULTS: Ingestion of EGb 761 produced no significant changes in diagnostic laboratory tests in either group, except reducing glycosylated hemoglobin A(1c) levels (from 7.7+/-1.2 to 7.2+/-0.9%, P<0.05) in T2DM the subjects. The pharmacokinetic parameters of metformin were all significantly different (P<0.05) between the NGT (500 mg) and 8 out of 10 of the T2DM subjects who were prescribed 500 mg of metformin during the placebo cycles. During the EGb 761 cycles, only the elimination half-life in the T2DM subjects was significantly increased (0.117+/-0.085 to 0.141+/-0.100, P<0.05). CONCLUSION: The co-ingestion of 120 mg of EGb 761 and 500 mg of metformin did not significantly affect the pharmacokinetic properties of metformin. Further studies are required to verify this observation for smaller and larger dose of metformin with other doses of EGb 761, since T2DM patients on medication constitute a very heterogeneous group. PMID- 16698135 TI - Maximum acceptable infusion rate of citrate: relationship between blood ionized calcium levels and cardiovascular effects in anesthetized rats. AB - BACKGROUND & AIMS: Citrate is a useful chemical as a stabilizer for infusion solutions. However, cardiovascular depression associated with ionized hypocalcemia has been observed during massive transfusion of citrated blood products. The goal of the present study was to determine the maximum acceptable infusion rate of citrate and safe blood ionized calcium (Ca(2+)) levels. METHODS: Citrate was administered intravenously to anesthetized rats at infusion rates between 0.5 and 2.0 mmol/kg/h for 4 h. Changes in heart rate (HR), arterial blood pressure, and the concentrations of plasma citrate and blood Ca(2+) were measured. RESULTS: Infusion of citrate caused decreases in arterial blood pressure and HR, but no severe cardiovascular depression was observed at infusion rates up to 1.0 mmol/kg/h. Plasma citrate levels reached a steady state within 1 h after the start of infusion at up to 1.0 mmol/kg/h. The concentrations of plasma citrate and blood Ca(2+) were 1.35 and 0.89 mmol/l, respectively, 4h after the start of infusion at 1.0 mmol/kg/h. CONCLUSIONS: The maximum acceptable infusion rate of citrate was 1.0 mmol/kg/h in anesthetized rats, and no severe cardiovascular effects were observed when the blood Ca(2+) level was 0.89 mmol/l or above. PMID- 16698136 TI - Comparison of tolerance and change of intragastric pH between early nasogastric and nasojejunal feeding following resection of colorectal cancer. AB - BACKGROUND: Early feeding after injury has been suggested to decrease morbidity and mortality in many studies. Intrajejunal feeding has been preferred over intragastric feeding due to earlier return of peristalsis following laparotomy. Few reports, however, have focused on the tolerance and change in pH inside the stomach after intragastric and intrajejunal feeding. The aim of the present study was the assessment of (1) the postoperative tolerance of intragastric and intrajejunal feeding, and (2) the effect of intragastric and intrajejunal feeding on intragastric pH value. MATERIALS AND METHODS: From April 1998 to October 2002, 140 patients underwent colon resection for colorectal cancer entered the study. The patients were divided into seven groups of 20 patients each. Group I was kept on NPO for 1 week. Groups II, III, and IV were fed through a nasogastric (NG) tube from the second to the sixth postoperative day (POD) with low residual (Osmolite-HN), high-fat (Pulmocare), and glutamine-containing (AlitraQ) enteral formulas, respectively. Groups V, VI, and VII were fed through a nasojejunal (NJ) tube from the second to the sixth POD with Osmolite-HN, Pulmocare, and AlitraQ, respectively. Feeding started at 500 kcal/500 cm(3)/d. If the patient tolerated the formula well, feeding increased to 1,500 kcal/1,500 cm(3)/d the following day. Intragastric pH was measured preoperatively and then twice daily until the sixth POD. RESULTS: Poor tolerance occurred in 14 patients (23%) with NG tube feeding and 18 patients (30%) with NJ tube feeding. The pH value of intragastric juice increased significantly once NG feeding started (3.67+/-1.33 on the third POD; 4.28+/-1.26 on the sixth POD). However, the pH value remained low after NJ feeding was started (2.09+/-1.46 on the third POD; 2.14+/-1.49 on the sixth POD). CONCLUSIONS: This series suggests that (1) the majority of patients can tolerate early feeding well following resection of colorectal cancer, and NJ feeding is not necessarily better tolerated than NG feeding; (2) early NG, but not the NJ feeding, can significantly elevate the intragastric pH value in patients who underwent resection of colorectal cancer. NG may be more effective than NJ feeding in preventing stress-induced gastropathy by elevating the pH value of intragastric juice. PMID- 16698137 TI - A method for implementation of nutritional therapy in hospitals. AB - BACKGROUND AND AIMS: Many barriers make implementation of nutritional therapy difficult in hospitals. In this study we investigated whether, a targeted plan made by the staff in different departments could improve nutritional treatment within selected quality goals based on the ESPEN screening guidelines. METHODS: The project was carried out as a continuous quality improvement project. Four different specialities participated in the study with a nutrition team of both doctors, nurses, and a dietician, and included the following methods: (1) Pre measurement: assessment of quality goals prior to study including the use of screening of nutritional risk (NRS-2002), whether a nutrition plan was made, and monitoring was documented in the records. (2) INTERVENTION: multidisciplinary meeting for the ward staff using a PC-based meeting system for detecting barriers in the department concerning nutrition, elaboration of an action plan and implementation of the plan. (3) Re-measurement: as in (1) based on information from records and patient interviews, and an evaluation based on focus group interview with the staff. Patients who gave informed consent to participate in the study (>14 years) were included consecutively. Mann-Whitney and Kruskal Wallis test was used for ordinal data, and Pearson chi(2) test for nominative data. P values <0.05 were considered significant. The study was performed in accordance with the Research Ethics Committee. RESULTS: In this study 141/122 patients were included before/after the implementation period with a mean weight loss within the last 3 months of 6.2 and 5.2 kg, respectively. Before the study we found that BMI was not measured. More than half of the patients had a weight loss within the last 3 months, and 40% had a weight loss during hospitalization, and this was not documented in the records. About 75% had a food intake less than normal within the last week, and nearly one-third were at a severe nutritional risk, and only 33% of these had a nutrition plan, and 18% a plan for monitoring. Barriers concerning nutrition included low priority, no focus, no routine or established procedures, and insufficient knowledge, lack of quality and choice of menus, and lack of support from general manager of the hospital. The staff introduced individually targeted procedures including assigning of responsibility, a nutrition record, electronic calculator of energy intake, upgrading of the dieticians and special diets, communication, and educational programs. A great consistency existed between barriers for targeted nutrition effort and ideas for improvement of the quality goals between the different departments. Quality assessment after study showed an overall significant improvement of the selected quality goals. CONCLUSION: The introduction of a new method for implementation of nutritional therapy according to ESPEN screening guidelines seems to improve nutritional therapy in hospitals. The method included assessment of quality goals, identification of barriers and individual targeted plans for each department followed by an evaluation process. The model has to be refined further with relevant clinical endpoints. PMID- 16698138 TI - Longitudinal trends in quality of life after starting home parenteral nutrition: a randomised controlled study of telemedicine. AB - BACKGROUND AND AIMS: This study defines and quantifies longitudinal changes in quality of life (QoL) at the time of first discharge home on home parenteral nutrition (HPN) and over the first year. METHODS: Results were compared in patients in standard contact with a nutrition nurse specialist by telephone, with results of those in contact via telemedicine in a randomised controlled trial. Participants were recruited from nine UK HPN centres. Patients were randomised to receive telemedicine upon initial discharge or after 1 year. The SF36 was the principal instrument chosen to determine QoL throughout the year on three predetermined occasions. EQ5D and hospital anxiety and depression scores were also recorded. RESULTS: Thirty participants were recruited to the study from March 2001 to June 2003. In all domains, QoL scores were significantly lower than normative data at discharge. QoL scores significantly improved over the first 6 months in physical functioning, physical role (RP), vitality (VT), social functioning (SF), emotional-role (RE) domains, and mental component summary (MCS). At 6 months RE, mental health (MH) and MCS were no longer significantly lower than normative data. There was no significant change in bodily pain (BP), general health (GH), MH, and physical component summary (PCS). Opiate use significantly reduced SF36 domains RP, BP, VT, SF, MH, and MCS at 6 months and was associated with more subsequent inpatient episodes and central line reinsertions. Patients with an acute onset of intestinal failure had less pain and better GH scores at 6 months, and had less inpatient episodes after discharge than patients with a more chronic onset. Telemedicine had no impact on QoL or subsequent clinical outcome. CONCLUSIONS: Aspects of QoL improve over the first 6 months of HPN. Opiate use and chronic diagnosis have a negative impact on some elements of QoL and clinical outcome variables. PMID- 16698139 TI - Body composition and echocardiographic abnormalities associated to anemia and volume overload in heart failure patients. AB - BACKGROUND: Heart failure (HF) patients with hemodilutional anemia had a worse prognosis than those with true anemia. Volume overload (VO) in anemic patients must be identified in order to select the best treatment. METHOD: We assessed the prevalence of anemia and VO in systolic and diastolic HF patients, and its effects on clinical status, body composition, and quality of life (QOL). Anemia was defined according to the OMS criteria and VO was assessed by bioelectrical impedance analysis (BIA). Echocardiography variables were also obtained. RESULTS: We studied 132 HF patients, 59.1% with systolic and 40.9% with diastolic HF. Anemia was present in 29.5% patients with systolic and 24.1% with diastolic HF (P=0.7). Anemic patients in the systolic HF group tend to be worse functional class than the non-anemic subjects; moreover, in both groups of cardiac dysfunction, the anemic patients had less grip strength, lower body mass index (BMI) and waist and hip circumferences, and a higher percentage of extracellular water (ECW), a higher proportion of patients with total serum cholesterol concentration below the 25th percentile, and a lower serum albumin. When patients were divided in 4 subgroups according to the presence of anemia and/or VO, we observed that VO affected the body composition not only through lower BMI, and waist and hip circumferences but also through a lower phase angle. Systolic HF patients with anemia and VO had the highest dimension of left atrium and pulmonary artery pressure (PAP) levels. CONCLUSION: Anemia is common in HF and its prevalence is similar in systolic and diastolic HF patients. Anemia was associated to VO, less grip strength and body composition abnormalities; however, VO was related to more severe body composition and echocardiographic abnormalities, as well as to an impairment of QOL. So that, it is important to differentiate between those who have anemia associated to VO and those who have true anemia. PMID- 16698140 TI - Validation of the VO2000 calorimeter for measuring resting metabolic rate. AB - BACKGROUND & AIMS: Metabolic carts used in laboratory settings for the measurement of resting metabolism are cumbersome limiting their use in the field. The validity of a newly developed portable calorimeter (Medical Graphics VO2000) under resting conditions was assessed in comparison to a well-established reference system, the DELTATRAC. METHODS: Gas exchange and energy expenditure were measured for 25 min consecutively using the two devices. Values of the last 20 min were averaged and used in the analysis. The order of device for the first subject was randomly chosen and the calorimeters were alternated thereafter. RESULTS: Among 33 subjects, acceptable measures of resting metabolism were obtained in 25 (11 men) aged 20-78 years because eight subjects (three men) either hyperventilated or did not adapt well enough to the facemask. VO(2), VCO(2), and RQ were not significantly different between devices. Small (2.8%) non clinically relevant mean differences (-0.145+/-0.341 MJ day(-1)) were found. Results of the two devices were highly correlated (r=0.95) yielding a more accurate estimate than predictive equations. CONCLUSIONS: The VO2000 calorimeter is a valid system to measure resting metabolism but the facemask may not be suitable for some people. PMID- 16698141 TI - Measurements of gastric emptying during continuous nasogastric infusion of liquid feed: electric impedance tomography versus gamma scintigraphy. AB - BACKGROUND & AIMS: Continuous nasogastric infusion is commonly used to deliver enteral feed but current methods used to assess tolerance based on aspiration and measurement of gastric residual volume have been criticised. Electric impedance tomography (EIT) measures gastric emptying by monitoring changes in epigastric impedance when a meal progressively empties from the stomach. AIMS: (1) to establish whether EIT was a valid method for measuring gastric emptying during continuous nasogastric infusion by comparing it with gamma scintigraphy (GS) and (2) to provide data on gastric emptying patterns during continuous nasogastric infusion. METHODS: Gastric emptying of 400 ml of enteral feed given over 200 min was measured simultaneously using EIT and GS in 10 healthy volunteers (five male and five female). RESULTS: Gastric emptying curves were obtained in 10 subjects by EIT but only eight by GS. Visual examination of the curves showed reasonable agreement. Patterns of emptying and filling during continuous nasogastric infusion were variable between individuals; the prevailing pattern was a trend towards a steady-state volume of approximately 50-125 ml. CONCLUSIONS: While EIT does not provide an accurate estimate of gastric volume during continuous infusion, it does show patterns of gastric emptying over time. With further development this could make it a useful tool for monitoring gastric emptying in patients at risk of gastroparesis. PMID- 16698142 TI - An in vivo and in vitro study of erythrocyte membrane acetylcholinesterase, (Na+, K+)-ATPase and Mg2+-ATPase activities in basketball players on alpha-tocopherol supplementation. The role of L-carnitine. AB - BACKGROUND & AIMS: Vitamin E (alpha-tocopherol, alpha-Te) and carnitine reduce lipid peroxidation. THE AIM WAS TO: To investigate the erythrocyte membrane acetylcholinesterase (AChE), Na+, K+-ATPase and Mg2+-ATPase activities in basketball players with or without alpha-Te supplementation, before and after training. In vitro, we aimed to find out any additional effect of L-carnitine (L C) on the modulated enzyme activities. SUBJECTS AND METHODS: Blood was obtained from 10 players before (group A), after exercise (group B) and after 1 month on alpha-Te (200 mg/24 h orally) supplementation before (group C) and after the game (group D). Lactate, pyruvate, muscle enzyme activities and total antioxidant status (TAS) were measured with commercial kits. Catecholamines and alpha-Te were determined with HPLC methods and membrane enzyme activities spectrophotometrically. RESULTS: Lactate, pyruvate, muscle enzymes and catecholamine levels were increased (P<0.001) in all groups after training. Alpha Te levels and Mg2+-ATPase activity remained unaltered before and after exercise. TAS was decreased in the groups after the game. AChE activity was increased in group B (P<0.01) and decreased in group D (P<0.01). After the exercise, Na+, K+ ATPase activity was increased in group B and remained unaltered in group D. In vitro incubation of membranes from group D with L-C resulted in a partially restoration of the membrane AChE activity, whereas Na+, K+-ATPase and Mg2+-ATPase activities were found unchanged. CONCLUSIONS: Alpha-Te supplementation in basketball players results in an increase of TAS and AChE activity, whereas the other enzyme activities were found unchanged. L-C addition may restore AChE activity, which was modulated by training in players on alpha-Te. PMID- 16698143 TI - The effect of different macronutrient infusions on appetite, ghrelin and peptide YY in parenterally fed patients. AB - BACKGROUND & AIMS: Patients receiving parenteral nutrition (PN) still feel hungry despite adequate provision of calories intravenously. It is not known whether PN or its constituent macronutrients acutely affect appetite and to what degree this may be mediated by ghrelin and peptide YY (PYY). METHODS: Six medically stable patients (four men) with intestinal failure receiving PN received an isocaloric 200 kcal infusion on three separate occasions following a 12 h fast. The infusions consisted of either carbohydrate (10% dextrose), fat (10% intralipid) or mixed protein/carbohydrate (PN). Changes in ghrelin and peptide YY levels and changes in subjective symptoms of hunger, satiety and nausea during each macronutrient infusion were assessed. RESULTS: None of the three infusions acutely affected subjective symptoms of hunger, satiety and nausea (P>0.05 ANOVA). Ghrelin levels decreased significantly during dextrose [-19.1 (-35.9, 12.4), regression coefficient (95% CI), P<0.001] and parenteral nutrition infusions [-18.2 (-26.8, -9.6), P<0.001]. Lipid infusion had no effect on ghrelin levels but led to a significant decrease in PYY [-0.076 (-0.0123, -0.028), P=0.004]. Dextrose and PN infusion had no significant effect on PYY levels. CONCLUSIONS: Dextrose and PN infusions decrease ghrelin levels. Lipid infusion does not affect ghrelin levels but in contrast to oral nutrients leads to a significant decrease in PYY. Despite these changes, in patients receiving PN, macronutrient infusions do no acutely affect appetite. PMID- 16698144 TI - Fecal energy losses in enterally fed intensive care patients: an explorative study using bomb calorimetry. AB - BACKGROUND & AIMS: Early enteral nutrition and tailored supply of nutrients have become standard in most of the intensive care units (ICU). So far little attention has been given to losses of energy in the stools. The purpose of this explorative study was to evaluate the energy losses of patients with loose stools, necessitating the use of a feces-collector device in a tertiary academic ICU. METHODS: In a group of 13 fully enterally fed and mechanically ventilated patients with loose stools, the daily energy loss in feces was determined, using bomb calorimetry. Malabsorption was defined as an absorption capacity of 85% or less. Energy expenditure was determined with indirect calorimetry. RESULTS: Six out of 13 (46%) patients fulfilled the criterion of malabsorption. The mean total energetic absorption capacity was 84.6+/-13.3%. The mean capacity of absorption of fat was 89.7+/-16.3%. The caloric value of energy loss had a mean of 301+/-259 kcal/day. Fecal fat loss proved not to be a good indicator of total fecal energy loss. A total of 4/13 patients (31%) had a net negative energy balance of over 500 kcal/day. A daily feces production of 250 g or more was a good predictor of malabsorption. Energy loss could accurately be predicted by using a factor 4.87 for the combined energetic value of protein and carbohydrates, if dry weight and fecal fat content are known. CONCLUSIONS: In this clinical study on ICU patients with loose stools, malabsorption proved to be a frequently occurring and so far unrecognized problem, contributing strongly to negative energy balances in 1/3 of the patients. PMID- 16698145 TI - Metabolic syndrome: analogies with subtypes of protein energy malnutrition. PMID- 16698146 TI - Association of abdominal obesity with decreased serum levels of carotenoids in a healthy Japanese population. AB - BACKGROUND & AIMS: Few epidemiological studies have investigated the relationship between abdominal adiposity and serum levels of carotenoids. This study investigated the relationship between total and abdominal obesity and serum levels of carotenoids. METHODS: Study population consisted of healthy Japanese males (n=192) and females (n=398) who attended a health examination screening (age: 49-86 years). Serum carotenoids were measured by high-performance liquid chromatography. Multiple logistic regression analysis was used to provide the multivariate-adjusted odds ratio (OR) and 95% confidence interval (CI) of obesity indices for low levels of serum carotenoids. RESULTS: In females, High waist circumference was associated with low levels of serum canthaxanthin (OR=2.00, 95% CI=1.26-3.19) and beta-carotene (BC) (OR=2.01, 95% CI=1.23-3.28). High waist hip ratio was also associated with low levels of serum alpha- carotene and BC (OR=2.44, 95% CI=1.49-4.03, and OR =1.97, 95% CI=1.20-3.28, respectively). Similar results were obtained when body mass index was added to confounding factors. In males, however, there were no significant associations between obesity indices and serum levels of carotenoids. CONCLUSIONS: Our results suggest that abdominal fat accumulation is associated with oxidative stress as determined by low levels of serum carotenoids in females. In males, further study excluding the effect of strong confounding factors, including smoking, is needed. PMID- 16698147 TI - ESPEN Guidelines on Enteral Nutrition: Wasting in HIV and other chronic infectious diseases. AB - Undernutrition (wasting) is still frequent in patients infected with the human immunodeficiency virus (HIV), despite recent decreases in the prevalence of undernutrition in western countries (as opposed to developing countries) due to the use of highly active antiretroviral treatment. Undernutrition has been shown to have a negative prognostic effect independently of immunodeficiency and viral load. These guidelines are intended to give evidence-based recommendations for the use of enteral nutrition (EN) by means of oral nutritional supplements (ONS) and tube feeding (TF) in HIV-infected patients. They were developed by an interdisciplinary expert group in accordance with officially accepted standards and is based on all relevant publications since 1985. Nutritional therapy is indicated when significant weight loss (>5% in 3 months) or a significant loss of body cell mass (>5% in 3 months) has occurred, and should be considered when the body mass index (BMI) is <18.5 kg/m(2). If normal food intake including nutritional counselling and optimal use of ONS cannot achieve an adequate nutrient intake, TF with standard formulae is indicated. Due to conflicting results from studies investigating the impact of immune-modulating formulae, these are not generally recommended. The results obtained in HIV patients may be extrapolated to other chronic infectious diseases, in the absence of available data. PMID- 16698148 TI - The effect of green tea in oxidative stress. AB - BACKGROUND & AIMS: Green tea, an infusion prepared with the leaves of Camellia sinensis is particularly rich in flavonoids, which are strong antioxidants. Tea drinking, by providing antioxidants, may become valuable in several oxidative stress conditions. Our aim was to evaluate the effect of green tea drinking on some factors reflecting the development of oxidative stress in plasma and in erythrocytes. METHODS: The study was performed in 34 Portuguese subjects. We evaluated the total antioxidant status (TAS), the lipid peroxidation products malonyldialdehyde (MDA) and malonyldialdehyde+4-hydroxy-2(E)-nonenal (MDA+4-HNE) and the oxidative changes in erythrocyte membrane, namely membrane bound haemoglobin (MBH) and the band 3 profile. Analytical evaluations were performed after 3 weeks drinking 1l of water daily, and after 4 weeks drinking 1l of green tea daily. Tea was prepared daily at the same conditions of temperature, time of infusion and concentration. RESULTS: After green tea drinking, we found a significant reduction in serum levels of MDA and MDA+4-HNE and in the oxidative stress within the erythrocyte, as suggested by a significantly lower value of MBH and by changes in band 3 profile towards a normal mean profile, namely an increase in the band 3 monomer. A rise in the antioxidant capacity was also observed. CONCLUSIONS: Our data suggest for green tea drinking a beneficial effect, by reducing the development or the enhancement of oxidative stress and, therefore, protecting the individual for oxidative stress diseases. Moreover, we propose further studies about the value of band 3 profile and of MBH in providing a cumulative measurement of the effect of green tea drinking upon the oxidative stress in cells. Moreover, further studies are also needed, to clarify the effect of green tea consumption, the value of regular green tea consumption and the way it should be prepared to reach a healthy effect. PMID- 16698149 TI - Monitoring of patients on home parenteral nutrition (HPN) in Europe: a questionnaire based study on monitoring practice in 42 centres. AB - INTRODUCTION & AIM: To gather information about how adult patients on home parenteral nutrition (HPN) are monitored after discharge from the hospital. METHOD: A questionnaire about HPN monitoring practice was circulated to HPN centres in eight European countries through the representative of the ESPEN HAN working group. Centres were asked about guidelines, home visits and how monitoring and handling of complications were managed. RESULTS: Fourty-two centres in eight European countries completed the questionnaire: UK n=14, France n=9, Belgium n=4, Italy n=4, Poland n=4, Denmark n=4, Spain n=2, Germany n=1. The HPN experience of the centres was in the range 2-30 years. Centres ranged in size from 0 to 125 HPN patients representing a total number of 934 of whom 54% had received HPN for more than 2 years. The primary disease was non-malignant in 90% whilst 10% had been diagnosed with active cancer. Of the centres 92% had a HPN team and 66% had written guidelines for monitoring HPN. Home visits after discharge for monitoring purposes were carried out by 31 of the centres involving the HPN team, general practitioner, community nurse or home care agency. Stable patients on HPN for more than 12 months were monitored at the discharging hospital (73%), at a local hospital (12%), by the General Practitioner (11%) or by a home care agency (4%). Of the centres, 90% reported that the main responsibility for monitoring was assigned to a specific person. The intervals between monitoring visits for the stable HPN patient was in the range 1-6 months, 52% of the centres reported intervals of 2-3 months. In case of complications 76% of centres reported that patients got in touch with the HPN team, 2% the local hospital, 5% the home care agency, and 17% other. Re-admission to hospital was usually to the HPN centre and only occasionally to a local hospital. CONCLUSION: In Europe a specialised team at the discharging hospital monitors HPN patients and 66% of the centres had some kind of written guidelines. PMID- 16698150 TI - Anti-inflammatory potential of the probiotic dietary supplement Lactibiane Tolerance: in vitro and in vivo considerations. AB - BACKGROUND & AIMS: Probiotics are live microorganisms which when administered in adequate amounts confer a health benefit on the host. In inflammatory bowel disease (IBD), where major modifications of the intestinal microflora have been reported, there is an increasing interest in modulating the flora with probiotic products. This work addresses the anti-inflammatory potential of Lactibiane Tolerance, a probiotic dietary supplement (mixture of four strains) using in vitro and in vivo approaches. METHODS: Comparison of the four individual strains and the commercial product reconstituted from them was conducted by in vitro tests (cytokine release after 24h stimulation of human peripheral blood mononuclear cells (PBMC)). The potential immunomodulatory characteristics of Lactibiane Tolerance were determined in vivo in an acute mice model of 2,4,6 trinitrobenzene sulfonic acid (TNBS)-induced colitis. Assessment of colitis included blinded histological and macroscopic scores. RESULTS: We showed that Lactibiane Tolerance has anti-inflammatory properties in vitro by stimulating IL 10 production and in vivo by conferring a significant protective effect in the TNBS-induced colitis model (more than 50% decrease of colitis symptoms, P<0.01). CONCLUSIONS: These results demonstrate that a probiotic dietary supplement, Lactibiane Tolerance, can significantly prevent the initial injury of TNBS and could stimulate the initiation of clinical trials in IBD. PMID- 16698151 TI - Intestinal anti-inflammatory activity of combined quercitrin and dietary olive oil supplemented with fish oil, rich in EPA and DHA (n-3) polyunsaturated fatty acids, in rats with DSS-induced colitis. AB - BACKGROUND AND AIMS: Previous studies have described the intestinal anti inflammatory effects exerted by the bioflavonoid quercitrin (QR) and by an n-3 polyunsaturated fatty acids (PUFA)-enriched diet in experimental models of rat colitis. The aim of the present study was to test if the combination of both treatments would result in an improvement in the intestinal anti-inflammatory effect achieved separately. METHODS: Colitis was induced in female Wistar rats by incorporating dextran sodium sulfate (DSS) in drinking water at 5% (w/v) for 5 days and at 2% (w/v) for the following 10 days. Five groups of rats (n=10) were used: two of them received an olive-oil-based diet with fish oil, rich in n-3 PUFA (FO diet) for 2 weeks before colitis induction and until the end of the experiment, and one of those also was administered daily QR (1mg/kg, PO), starting when DSS concentration was changed. DSS colitis was induced in other two groups fed with standard rat diet, one of them being administered QR as before. A non-colitic group fed standard diet was also included. After that period, the rats were sacrificed and colonic damage was assessed both histologically and biochemically. RESULTS: The concurrent administration of FO diet and QR exhibited an intestinal anti-inflammatory effect, as evidenced by a significant improvement of all biochemical parameters of colonic inflammation assayed in comparison with non-treated colitic rats. Thus, both colonic myeloperoxidase (MPO) and alkaline phosphatase (AP) activities were significantly reduced compared with untreated colitic rats. In addition, a complete restoration of colonic glutathione content, which was depleted as a consequence of the colonic insult, was obtained in rats treated with QR plus FO diet; this content was even higher than that obtained when colitic rats were treated with FO diet alone. When compared with the control colitic group, the combined treatment was also associated with a lower colonic nitric oxide synthase and cyclooxygenase-2 expression as well as with a significant reduction in different colonic proinflammatory mediators assayed, i.e. leukotriene B(4), tumor necrosis factor alpha and interleukin 1beta, showing a significantly greater inhibitory effect of the latter in comparison with rats receiving FO diet without the flavonoid. CONCLUSIONS: These results support the potential synergism between the administration of the flavonoid and the incorporation of olive oil and n-3 PUFA to the diet for the treatment of these intestinal inflammatory disorders. PMID- 16698152 TI - ESPEN Guidelines on Enteral Nutrition: Surgery including organ transplantation. AB - Enhanced recovery of patients after surgery ("ERAS") has become an important focus of perioperative management. From a metabolic and nutritional point of view, the key aspects of perioperative care include: Enteral nutrition (EN) by means of oral nutritional supplements (ONS) and if necessary tube feeding (TF) offers the possibility of increasing or ensuring nutrient intake in cases where food intake is inadequate. These guidelines are intended to give evidence-based recommendations for the use of ONS and TF in surgical patients. They were developed by an interdisciplinary expert group in accordance with officially accepted standards and are based on all relevant publications since 1980. The guideline was discussed and accepted in a consensus conference. EN is indicated even in patients without obvious undernutrition, if it is anticipated that the patient will be unable to eat for more than 7 days perioperatively. It is also indicated in patients who cannot maintain oral intake above 60% of recommended intake for more than 10 days. In these situations nutritional support should be initiated without delay. Delay of surgery for preoperative EN is recommended for patients at severe nutritional risk, defined by the presence of at least one of the following criteria: weight loss >10-15% within 6 months, BMI<18.5 kg/m(2), Subjective Global Assessment Grade C, serum albumin <30 g/l (with no evidence of hepatic or renal dysfunction). Altogether, it is strongly recommended not to wait until severe undernutrition has developed, but to start EN therapy early, as soon as a nutritional risk becomes apparent. PMID- 16698153 TI - CLA and n-3 PUFA differentially modulate clinical activity and colonic PPAR responsive gene expression in a pig model of experimental IBD. AB - BACKGROUND AND AIMS: Conjugated linoleic acid (CLA) and n-3 polyunsaturated fatty acids (PUFA) have been proposed as important pharmaco-nutrients for modulating mucosal immunity and therapeutic responses in patients with inflammatory bowel disease (IBD). We evaluated the ability of CLA and n-3 PUFA alone or in combination to modulate IBD in a pig model of dextran sodium sulfate (DSS) induced colitis. METHODS: Sixty-four, 15-day-old pigs were used to evaluate the effect of CLA, n-3 PUFA and a 50:50 mixture of CLA and n-3 PUFA on growth, clinical activity and colonic PPAR-responsive gene expression. Diets were formulated to contain: 1.33% soybean oil (control); 1.33% CLA; 1.33% fish oil; or 1.33% of a 50:50 mixture of CLA and fish oil. Intestinal inflammation was induced by an intragastric challenge with DSS on day 42 of dietary supplementation. The colonic expression of peroxisome proliferator-activated receptor-gamma (PPAR gamma), PPAR gamma- and delta-responsive genes, keratinocyte growth factor (KGF) and tumor necrosis factor (TNF-alpha) were assayed by quantitative RT-PCR. RESULTS: The onset of IBD was delayed, colitis less severe and growth suppression attenuated in pigs fed CLA, which correlated with induction of colonic PPAR gamma and its responsive gene PPAR gamma-coactivator-1alpha (PGC1-alpha) and downregulation of TNF-alpha. However, dietary supplementation with n-3 PUFA alone or in combination with CLA resulted in an early onset of disease (i.e., day 2) and faster recovery on days 6 and 7, which correlated with a marked induction of the PPAR delta-responsive gene uncoupling protein 3 (UCP3). CLA and n-3 PUFA acted synergistically to upregulate colonic KGF expression in DSS-challenged pigs but n-3 PUFA blocked CLA-induced PPAR gamma activation. CONCLUSION: Dietary CLA supplementation upregulated colonic PPAR gamma expression and contributed to delaying the onset of experimental IBD, whereas n-3 PUFA failed to protect from IBD, although it accelerated colonic regeneration and clinical remission by activating PPAR delta. PMID- 16698154 TI - The conduct of ethical research collaboration across international and culturally diverse communities. AB - OBJECTIVE: to present recently developed guidelines by the International Confederation of Midwives (ICM) to guide the ethical conduct of research by midwives across international and culturally diverse communities. To illustrate the complexities of conducting collaborative research across diverse communities, a case study is provided highlighting unequal power balances, legal, ethical and cultural constraints. BACKGROUND: the research guidelines are structured using a human rights framework. The document was developed with input and consideration by the ICM Research Standing Committee and comments of 35 delegates who participated in a workshop to evaluate the guidelines from the perspectives of frontline midwifery clinicians, leaders and researchers. It is intended to serve as a blueprint for midwives aiming to enter into a collaborative research partnership with ethnic and culturally diverse groups. CONCLUSIONS: although the document was developed with the international midwifery community in mind, it will guide any researcher seeking to conduct research in the best interest of women, their newborn babies, their families, and their wider communities. PMID- 16698156 TI - Trace metal enrichments in core sediments in Muthupet mangroves, SE coast of India: application of acid leachable technique. AB - Core sediments from Mullipallam Creek of Muthupet mangroves on the southeast coast of India were analyzed for texture, CaCO(3), organic carbon, sulfur and acid leachable trace metals (Fe, Mn, Cr, Cu, Ni, Co, Pb, Zn and Cd). Textural analysis reveals a predominance of mud while CaCO(3) indicates dissolution in the upper half of the core, and reprecipitation of carbonates in reduction zones. Trace metals are diagenetically modified and anthropogenic processes control Pb and, to some extent, Ni, Zn and Fe. A distinct event is identified at 90 cm suggesting a change in deposition. Strong relationship of trace metals with Fe indicates that they are associated with Fe-oxyhydroxides. The role of carbonates in absorbing trace metals is evident from their positive relationship with trace metals. Comparison of acid leachable trace metals indicates increase in concentrations in the study area and the sediments act as a sink for trace metals contributed from multiple sources. PMID- 16698157 TI - Primary care professionals' perceptions of depression in older people: a qualitative study. AB - An understanding of patients' perspectives is crucial to improving engagement with health care services. For older people who may not wish to bother medical professionals with problems of living such as depression, such exploration becomes critical. General practitioners (GPs), nurses and counsellors working in 18 South London primary care teams were interviewed about their perceptions of depression in older people. All three professional groups shared a predominantly psychosocial model of the causes of depression. While presentation of somatic symptoms was seen as common in all age groups, identification of depression in older patients was complicated by co-existent physical illnesses. GPs reported that older patients rarely mentioned psychological difficulties, but practice nurses felt that older people were less inhibited in talking to them about "non medical" problems. Many older people were perceived to regard symptoms of depression as a normal consequence of ageing and not to think it appropriate to mention non-physical problems in a medical consultation. Men were thought to be particularly reluctant to disclose emotional distress and were more vulnerable to severe depression and suicide. Some GPs had mixed feelings about offering medication to address what they believed to be the consequences of loneliness and social isolation. Participants thought that many older people regard depression as a "sign of weakness" and the perceived stigma of mental illness was widely recognised as a barrier to seeking help. Cultural variations in illness beliefs, especially the attribution of symptoms, were thought to profoundly influence the help-seeking behaviour of elders from minority ethnic groups. Families were identified as the main source of both support and distress; and as such their influence could be crucial to the identification and treatment of depression in older people. PMID- 16698158 TI - Education and health in 22 European countries. AB - This study investigates educational health inequalities in 22 European countries. Moreover, age and gender differences in the association between education and health are analysed. The study uses data from the European Social Survey 2003. Probability sampling from all private residents aged 15 years and older was applied in all countries. The European Social Survey includes 42,359 cases. Persons under age 25 were excluded to minimise the number of respondents whose education was not complete. Education was coded according to the International Standard Classification of Education. Self-rated health and functional limitations were used as health indicators. Results of multiple logistic regression analyses show that people with low education (lower secondary or less) have elevated risks of poor self-rated health and functional limitations. Inequalities are relatively small in Austria, Norway, Sweden, and the United Kingdom, large inequalities were found for Hungary, Poland, and Portugal. Analyses of age differences reveal that health effects of education are stronger at ages 25-55 than in the higher age groups. However, age differences in the education-health association vary between countries, sexes, and health indicators. In conclusion, our results confirm that educational inequalities in health are a generalised though not invariant phenomenon. Variations between countries, sexes and health indicators might be one explanation for the inconsistent results of other studies on age differences in the association between socioeconomic position and health. PMID- 16698159 TI - Women's health in mid-life: life course social roles and agency as quality. AB - Data from a prospective British birth cohort study showed that women who were childless, lone mothers or full-time homemakers between the ages of 26 and 53 were more likely to report poor health at age 54 than women who occupied multiple roles between these ages. To explain this finding we developed and tested a theory of role quality based on the concept of agency by drawing on Giddens' theory of structuration and Doyal and Gough's theory of human needs. According to our theory, the patriarchal structuration (drawing on Giddens' term) of work and family roles provides both limitation and opportunity for the expression of agency. Doyal and Gough's theory of human needs was then used to identify the restriction of agency as a possible influence on health. This theory of role quality was operationalised using a measure of work (paid and unpaid) quality at age 36 and a measure of work and family stress between ages 48 and 54. The relatively poor subjective health in mid-life of lone mothers was explained by work and family stress and adult social class. In contrast, the poor health in mid-life of long-term homemakers and childless women was less easily explained. Homemaker's excess risk of reporting poor health at age 54 remained strong and significant even after adjusting for role quality and socioeconomic indicators, and childless women were at an increased risk of reporting poor health despite the social advantage inherent in attaining educational qualifications and occupying professional or managerial occupations. This study highlights the need to develop measures of role quality specifically designed to capture agency aspects of social roles. PMID- 16698160 TI - A pilot clinical trial of oxcarbazepine in adults with attention-deficit hyperactivity disorder. AB - Despite the increasing recognition of attention-deficit hyperactivity disorder (ADHD) in adults, there is a paucity of controlled pharmacological trials. Recent reports have suggested the potential usefulness of mood stabilizing drugs for ADHD. To this end, the authors completed a pilot study with oxcarbazepine for the treatment of adults with ADHD. This was an open pilot study of oxcarbazepine (300 1,500 mg daily dosage) in adults who met DSM-IV criteria for ADHD. The treatment period was 8 weeks. Of the 9 subjects enrolled in the study (4 men, 5 women), 8 patients could be included in the analysis. At the endpoint of the active treatment, a significantly high proportion of subjects was considered improved while receiving oxcarbazepine. ADHD symptom checklist scores (ADHD-IV rating scale, Conners ADHD adult rating scale, ADHD self-rating [ADHD-SR] scale) showed significant reduction during the treatment period. Treatment with oxcarbazepine was relatively well tolerated; dizziness, sedation and nausea were the most frequently reported adverse effects. The results of this investigation indicate that oxcarbazepine may be a potentially useful agent for the treatment of ADHD in adults. However, placebo-controlled randomized trials are needed to provide evidence. PMID- 16698161 TI - Polysaccharide peptides from COV-1 strain of Coriolus versicolor inhibit tolbutamide 4-hydroxylation in the rat in vitro and in vivo. AB - Polysaccharide peptide (PSP), isolated from COV-1 strain of Coriolus versicolor, is commonly used as an adjunct in cancer chemotherapy in China. In this study, the effects of whole PSP extract and water extract of PSP on 4-hydroxylation of tolbutamide were investigated in rat liver microsomes in vitro and in vivo in the rat. Both the whole PSP extract and the water soluble fraction (0.5-20 microM) decreased the metabolism of tolbutamide to 4-hydroxytolbutamide in vitro. Enzyme kinetics studies showed that PSP inhibited tolbutamide 4-hydroxylase activity in a competitive, concentration-dependent manner. The whole PSP extract had a Ki value of 12.6 microM and IC50 at 18.4 microM, while the water extract had a Ki value of 6.9 microM and IC50 at 9.8 microM. Sulphaphenazole, a specific human CYP2C9 inhibitor, showed a Ki value of 30.8 microM and IC50 at 44.0 microM in the test system. In the pharmacokinetic studies in vivo, acute PSP (4 micromol/kg, i.p.) treatment did not produce significant changes in tolbutamide clearance, but produced a decrease in the Cinitial (7.4%) and an increase in the Vd (7.4%). Sub chronic pre-treatment of PSP (1-2 micromol/kg/day, i.p.) for three days did not affect the clearance and AUC of tolbutamide, but the Cinitial was decreased, together with increases in the T1/2, and Vd. The formation of 4 hydroxytolbutamide in vivo was decreased in both acute and sub-chronic studies. Taken together, this study demonstrated the PSP can inhibit tolbutamide 4 hydroxylation both in vitro and in vivo. Despite the fact that CYP isoforms that metabolise tolbutamide are different between rat and human liver due to different catalytic characteristics, and rat studies may not be directly extrapolatable to man, the concomitant use of PSP with other CYP2C substrates should be carefully monitored. PMID- 16698162 TI - Modulation of antipyrine clearance by polysaccharide peptide (PSP) isolated from Coriolus versicolor in the rat. AB - Polysaccharide peptide (PSP), isolated from Coriolus versicolor COV-1, has been previously shown to have immuno-stimulatory, anti-tumour and analgesic effects in animal models. When used as an adjunct in cancer chemotherapy in clinical trials carried out in China, PSP improved the quality of life in the patients by improving appetite and alleviating symptoms associated with cancer chemotherapy. In this study, the effects of non-toxic doses of PSP on phase I metabolism was investigated in the rat, using the conventional probe antipyrine. Acute PSP (3-5 micromol/kg, i.p.) treatment did not produce significant changes in antipyrine clearance. Sub-chronic treatment with PSP (1-3 micromol/kg/day, i.p., 3 days) decreased the antipyrine clearance (30-35%), with an increase in the plasma half life (T1/2) by 55% and an increase in the area under concentration-time curve (AUC) by 61%. Total hepatic cytochrome P450 (P450) was dose-dependently decreased (32-54%) after sub-chronic, but not the acute treatment of PSP. Given that PSP can affect phase I metabolism and hepatic cytochrome P450 content, the concomitant use of PSP with other therapeutic agents that undergo phase I metabolism should be carefully monitored. PMID- 16698163 TI - Long-lasting neurotoxicity of prenatal benzene acute exposure in rats. AB - Benzene is a common element of environmental pollution. Although this substance is not recognized as a teratogenic agent, it is not known whether prenatal exposure to benzene may induce neurobehavioral changes in the progeny. Benzene 0.1mg/kg was injected subcutaneously (s.c.) acutely at day 15 of gestation into pregnant female rats of the Sprague-Dawley strain and neurotoxicity of the substance was studied in pups and male adult animals of the same progeny. No change was found in total number of neonates, body weight and eye opening time between benzene-exposed animals and controls. No malformations were observed. At birth, neonatal reflexes (cliff aversion, forelimb placing, bar holding, forelimb grasping, startle) were scored in benzene-exposed pups and their percent appearance was found to be anticipated (more benzene-exposed pups exhibited reflexes each day) in comparison to that of control animals. Also, the completion (maximum appearance, i.e. 100% of the brood was found to exhibit each reflex) of neonatal reflexes in benzene-exposed animals preceded that of controls. Starting 2 months after birth, cognitive and motor performance was assessed only in male animals of the prenatally benzene-exposed progeny. The overall evaluation of motor activity in benzene-exposed animals in the open-field test revealed reduced ambulation in these rats as compared to control animals. Acquisition of active avoidance responses in the shuttle-box test, as assessed by the number of conditioned avoidance responses and the percent of learners, was impaired in benzene-exposed rats as compared to control animals. Prenatal exposure to benzene was also followed by reduced retention latency in a step-through passive avoidance task in two retention tests. These results suggest that acute exposure to benzene during gestational organogenesis may cause long-lasting changes in motor behavior and cognitive processes. This may be relevant for the assessment of benzene toxic profile for the progeny of pregnant subjects, although teratogenic effects are not observed. PMID- 16698164 TI - Reproducibility of skeletal muscle vasodilatation responses to Stroop mental challenge over repeated sessions. AB - Skeletal muscle blood flow responses to stress have implications for psychobiological disease pathways. An important assumption underlying psychophysiological studies relating stress reactivity with disease risk is that individuals are characterized by stable response profiles that can be reliably assessed using acute psychophysiological stress testing. We examined the reproducibility of forearm vasodilatation, blood pressure, and cardiac responses to a 2 min Stroop mental challenge over two repeated stress sessions that were on average 3.6 months apart. Participants were 21 healthy men and women (aged 21.8+/ 3.7 years). Vasodilatation, blood pressure and heart rate responses displayed no habituation between sessions, although there was significantly greater cardiac parasympathetic involvement during the second testing session. Significant test retest correlations between the sessions were observed for both forearm blood flow and heart rate reactivity. These findings demonstrate skeletal muscle vasodilatation responses to repeated stress are robust, so may be a useful psychophysiological indicator in studies of stress reactivity and disease risk. PMID- 16698165 TI - The use of an unpleasant sound as an unconditional stimulus in a human aversive Pavlovian conditioning procedure. AB - Ethical considerations can limit the use of traditional unconditional stimuli (US), such as electric shock and loud tones, when used in a human aversive Pavlovian conditioning procedure. The risk of the US causing pain or excessive anxiety is a particular concern when testing sensitive populations such as children, the elderly, and those with psychological or neurodevelopmental disorders. Two experiments used a differential conditioning procedure to determine whether an unpleasant sound (metal scraping on slate) could support the acquisition and extinction of conditioned responses to the same extent as either electric shock or a 100 dB(A) tone US. Experiment 1 (N=48) demonstrated equivalent or superior conditioning effects for the signal-based learning measures of US expectancy, skin conductance responses, and heart rate. Experiment 2 (N=57) yielded similar outcomes in the affective-based learning measures of startle blink modulation and pleasantness ratings. The results support the use of an unpleasant sound as a US in human Pavlovian conditioning experiments. PMID- 16698166 TI - Cross-sectional study of weight gain and increase in BMI throughout pregnancy. AB - OBJECTIVE: To generate reliable new reference ranges for weight gain and increase in body mass index (BMI) during pregnancy from a large population. STUDY DESIGN: In a prospective cross-sectional study at the Obstetric outpatient clinic, Zurich University Hospital, weight gain and BMI, before gestation and at the booking visit, were determined in 4034 pregnant women with accurately dateable singleton pregnancies (Caucasian: N = 3242, Asian (predominantly from Sri Lanka, Thailand and the Philippines): N = 578 and Black: N = 214). Women with known insulin dependent diabetes mellitus before pregnancy were excluded. Fifth, 50th and 95th centiles were presented for Caucasians and corresponding centile curves for Asians and Blacks. Simple and multiple regression analyses were performed for various risk factors. A significance level of P < 0.05 was used in all tests. RESULTS: Mean weight gain was 15.5+/-5.9 kg (34.2+/-13.0 lb) at term with values >25.4 kg (56.0 lb) and <5.7 kg (12.6 lb) for the 95th and the 5th centile, respectively. Mean BMI increased slightly and steadily to 28 kgm(-2) at term. Parity and pre-pregnancy BMI were significant determinants in Caucasians. Weight gain and BMI was slightly lower in Asians and Blacks. CONCLUSIONS: BMI centile curves have the advantage in that they consider height during the whole course of pregnancy. It may be an additional helpful tool in controlling weight gain in pregnancy. Further studies are required to determine the prognostic implications of values > or = 95th centile and < or = 5th centile. PMID- 16698167 TI - Enhancement kinetics of normal ovaries on dynamic contrast-enhanced MR imaging. AB - OBJECTIVE: The objective was to determine the enhancement kinetics of the normal ovaries in healthy women on dynamic contrast-enhanced magnetic resonance (DCE-MR) imaging. METHOD: Twenty-one women who had normal ovulatory cycles (volunteers; mean age=26.3 years, range=20-35), normal hormone profile, and apparently normal ovaries on ultrasonographic scan underwent DCE-MR imaging on a 1.5-T system (Philips Medical Systems, Gyroscan Intera, Best, The Netherlands) using a phased array pelvic coil. Sequential images with an imaging time of 14.4s per dynamic image were acquired before and after injection of a contrast bolus at 30s and at 1, 2, 3, 4, and 5 min. On postprocessing examination the following measurements were obtained for ovarian and muscle tissue: the signal intensity value per dynamic study, early phase enhancement rate, time to peak enhancement (Tp), and percentage of wash-out at the fifth minute were determined. Data of the ovaries and skeletal muscle were compared using Wilcoxon's rank sum test. RESULTS: Most of the mean values of the postcontrast signal intensity measurements, the mean values of the early phase enhancement rate, and the percentage of wash-out at the fifth minute were found to be significantly higher in ovary than in muscle (p<0.05). The mean values of Tp, on the other hand, were nonsignificantly lower in ovary than in muscle (p>0.05). On the examination of the mean signal intensity time data graphics the ovary showed a tendency toward greater and quicker enhancement and wash-out. CONCLUSION: In our opinion, DCE-MR imaging, which determines contrast enhancement such as wash-in and wash-out kinetics, can provide knowledge of ovarian vascularization. Thus, by using DCE-MR imaging, determination of these vascularization changes in various ovarian diseases may provide us with additional parameters in the diagnosis of and treatment strategies for ovarian diseases. PMID- 16698168 TI - Estrogen and phytoestrogens: Effect on eNOS expression and in vitro vasodilation in cerebral arteries in ovariectomized Watanabe heritable hyperlipidemic rabbits. AB - OBJECTIVES: To evaluate the effect of estrogen replacement therapy or soy isoflavones supplement on endothelium-dependent relaxation in vitro and gene expression of endothelial nitric oxide synthase (eNOS) in cerebral arteries in a rabbit model of human hypercholesterolemia. STUDY DESIGN: Thirty-six female ovariectomized Watanabe heritable hyperlipidemic (WHHL) rabbits were randomised to treatment with 17beta-estradiol (17beta-E(2)), SoyLife 150 or control for 16 weeks. Ring segments of basilar artery (BA) and posterior cerebral artery (PCA) were mounted in myographs for isometric tension recordings. Concentration response curves to carbamylcholine chloride, sodium nitroprusside (SNP) and l NAME were evaluated after precontraction with potassium. Total RNA was extracted, reverse transcribed and eNOS quantitated by real-time polymerase chain reaction (real-time PCR). RESULTS: Plasma cholesterol was significantly higher at termination in the SoyLife group (P<0.0001), whereas low-density lipoprotein (LDL) cholesterol was comparable in all treatment groups. Neither treatment influenced the endothelium-dependent responses to carbamylcholine chloride or l NAME or the endothelium-independent response to SNP in any of the arteries. Correspondingly, eNOS mRNA was similarly expressed in all treatment groups in both arteries. CONCLUSIONS: Improvement of cerebral endothelial function by estrogen or soy isoflavones in ovariectomized WHHL rabbits is not supported by the present data. The findings may be unique to the WHHL rabbit in which the hypocholesterolemic effect of estrogens mediated by upregulation of liver LDL receptors is excluded. PMID- 16698169 TI - Apoptosis pattern in human endometrium in women with pelvic endometriosis. AB - OBJECTIVE: In the present study we aimed to describe apoptosis patterns in eutopic endometrium in women suffering from endometriosis in order to assess its value as a marker of early forms of endometriosis, and also to try to answer whether endometriosis is caused by changes within the eutopic endometrium or whether endometriotic lesions change the characteristics of eutopic endometrium. STUDY DESIGN: The study was performed on 125 women treated in Division of Reproduction. In 52 patients peritoneal endometriosis was diagnosed (I(0)-23; II(0)-29). Seventy-three patients in whom no endometriotic foci could be found at laparoscopy were qualified as the control group. Endometrial biopsy 7-9 days after ovulation was taken for assessment of apoptosis (TUNEL) and routine histology. RESULTS: Apoptosis indices in the eutopic endometrium of women with endometriosis were lower compared to women without endometriosis. In the endometrial glands apoptosis indices were 2.94+/-1.66 versus 5.23+/-2.06 (p<0.01) in the group of women with and without endometriosis, respectively. In the endometrial stroma apoptosis indices were estimated at 2.04+/-1.72 in women with endometriosis and 4.12+/-1.62 in the control group (p<0.01). CONCLUSIONS: The observed changes could support the hypothesis of the different properties of eutopic endometrium in endometriotic women as a causing factor of peritoneal endometriosis. PMID- 16698170 TI - Use of chemotherapy for ovarian cancer during human pregnancy: case report and literature review. PMID- 16698171 TI - Reference curves of symphysis-fundus height in twin pregnancies. AB - OBJECTIVE: To generate reliable new reference ranges for symphysis-fundus height (SFH) in twin pregnancies using modern statistical methods and to evaluate whether small-for-gestational age (SGA) babies of women who had a SFH measurement after the 25th gestational week could be predicted by the SFH measurement in the reference curves and other maternal data. STUDY DESIGN: In a retrospective cross sectional study at the obstetric outpatient clinic, Zurich University Hospital, SFH was determined in 257 twin-pregnant women with accurately dateable twin pregnancies (Caucasians: N=217, Asians: N=15, Blacks: N=10, and 15 others). Exclusion criteria were intrauterine fetal death, and known fetal and maternal diseases, which influence SFH. Pregnant women with twins were divided in three groups according to the birth weight of the babies. Group I: both babies were appropriate for gestational age (AGA), group II: one baby was AGA and one SGA, and group III: both babies were SGA. RESULTS: SFH measurements increased linearly with gestational age (GA). The following rule of thumb is suggested for the 50th centile of SFH (cm)=gestational week+10% of gestational week. Age, height, weight and body mass index (BMI) before pregnancy, parity and ethnic group were insignificant determinants in SFH measurement. A prognostic score for identification of group III was created for a GA> or =25 weeks and BMI<30 kg/m2. CONCLUSIONS: Measuring SFH is simple, inexpensive and non-invasive and may be of some use for identifying twin mothers with SGA twin pairs. PMID- 16698172 TI - Ion solvation and water structure in potassium halide aqueous solutions. AB - The structure of water and the nature of ionic hydration is explored in aqueous solutions of potassium fluoride, chloride, bromide and iodide over a range of concentrations up to 4.8 ion pairs per 100 water molecules, using the combined techniques of neutron diffraction with hydrogen isotope substitution. The diffraction data are interpreted using the method of empirical potential structure refinement, which attempts to build a three-dimensional model of the scattering system consistent with the diffraction data. The water structure is strongly perturbed in the first hydration shells of both anion and cation, but is found to be only mildly perturbed outside of this region, with the largest effects occurring with the smallest anion and highest concentrations. For the potassium ion there are strong orientational correlations in the first hydration shell, with the water molecules lying with their dipole moments pointing almost directly away from the cation on average, but with an angular spread of approximately +/-60 degrees which is mildly dependent on the anion type present. For all the anions the water molecules in the first shell are strongly oriented with one O-H vector pointing directly towards the anion on average, with an angular spread of approximately +/-10 degrees for F(-), increasing to approximately +/-22 degrees for I(-). For both anions and cations the second hydration shell is much more disordered than the first, but there is a weak pattern of orientational correlation which becomes more pronounced with the larger anions. There is some evidence that the fluoride ion structures water significantly in its first hydration shell, but not beyond. The findings throw further light on recent findings that the orientational relaxation time for water outside the first shell of dissolved ions is the same as in the bulk liquid. PMID- 16698173 TI - The improvement of the algorithm for order parameter calculation (S2) from molecular dynamics simulation using the correlation motion function. AB - The generalized order parameter, S2, calculated from MD simulation trajectory using time-dependent internal Correlation Motion Function (CMF) agrees well with NMR derived S2 processed with the extended model-free analysis approach. However, the former lies considerably lower comparing to simple model-free derived data from NMR experiments. In the present study we analyze possible reasons of such disagreement. In the general case we propose to use preexponential factors from expression for internal CMF rather than ordinary S2 values. Particularly, in case of the simple model-free S(2) experimental values we suggest comparing them with S2(eff)=1+S2-Sf2 computed from MD simulation data. We show that the S2(eff) values are in a good agreement with NMR derived S2 values obtained using the simple model-free analysis. PMID- 16698174 TI - Managing mercury in the great lakes: an analytical review of abatement policies. AB - Mercury, a toxic metal known to have several deleterious affects on human health, has been one of the principal contaminants of concern in the Great Lakes basin. There are numerous anthropogenic sources of mercury to the Great Lakes area. Combustion of coal, smelting of non ferrous metals, and incineration of municipal and medical waste are major sources of mercury emissions in the region. In addition to North American anthropogenic emissions, global atmospheric emissions also significantly contribute to the deposition of mercury in the Great Lakes basin. Both the USA and Canada have agreed to reduce human exposure to mercury in the Great Lakes basin and have significantly curtailed mercury load to this region through individual and joint efforts. However, many important mercury sources, such as coal-fired power plants, still exist in the vicinity of the Great Lakes. More serious actions to drastically reduce mercury sources by employing alternative energy sources, restricting mercury trade and banning various mercury containing consumer products, such as dental amalgam are as essential as cleaning up the historical deposits of mercury in the basin. A strong political will and mass momentum are crucial for efficient mercury management. International cooperation is equally important. In the present paper, we have analyzed existing policies in respective jurisdictions to reduce mercury concentration in the Great Lakes environment. A brief review of the sources, occurrence in the Great Lakes, and the health effects of mercury is also included. PMID- 16698176 TI - Propiverine compared to oxybutynin in neurogenic detrusor overactivity--results of a randomized, double-blind, multicenter clinical study. AB - OBJECTIVES: To compare the efficacy and tolerability of propiverine and oxybutynin in patients with neurogenic detrusor overactivity. METHODS: Patients were eligible, if at least 18 years of age and suffering from neurogenic detrusor overactivity. Eligibility also required a maximum cystometric capacity less than 300 ml. After a one-week run-in period, propiverine 15 mg t.i.d. or oxybutynin 5mg t.i.d. were administered for 21 days. As primary efficacy outcomes urodynamic parameters were assessed. As tolerability outcome the percentage of patients with newly manifesting anticholinergic adverse events was taken. RESULTS: 131 patients were recruited at 20 study centers. The maximum cystometric capacity (ml) was increased significantly in the propiverine group from 198 (+/-110) to 309 (+/ 166), and in the oxybutynin group from 164 (+/-64) to 298 (+/-125). Similarly, maximum detrusor pressure during the filling phase (cm H(2)O) was lowered significantly in the propiverine group from 56.8 (+/-36.2) to 37.8 (+/-31.6), and in the oxybutynin group from 68.6 (+/-34.5) to 43.1 (+/-29.2). No significant differences resulted between treatment groups. Anticholinergic adverse events were reported less frequently in the propiverine compared to the oxybutynin group (63.0% versus 77.8%). Dryness of the mouth, the most frequent adverse event, was reported significantly less (47.1% versus 67.2%; p=0.02) in the propiverine compared to the oxybutynin group. CONCLUSION: Propiverine and oxybutynin are equally effective in increasing bladder capacity and lowering bladder pressure in patients with neurogenic detrusor overactivity. The trend for better tolerability of propiverine compared to oxybutynin achieved significance for dryness of the mouth. PMID- 16698177 TI - Presence of endocrine and exocrine markers in EGFP-positive cells from the developing pancreas of a nestin/EGFP mouse. AB - In order to purify and characterize nestin-positive cells in the developing pancreas a transgenic mouse was generated, in which the enhanced green fluorescent protein (EGFP) was driven by the nestin second intronic enhancer and upstream promoter. In keeping with previous studies on the distribution of nestin, EGFP was expressed in the developing embryo in neurones in the brain, eye, spinal cord, tail bud and glial cells in the small intestine. In the pancreas there was no detectable EGFP at embryonic day 11.5 (E11.5). EGFP expression appeared at E12.5 and increased in intensity through E14.5, E18.5 and post-natal day 1. Flow cytometry was used to quantify and purify the EGFP positive population in the E15.5 pancreas. The purified (96%) EGFP-expressing cells, which represent 20% of the total cell population, were shown by RT/PCR to express exocrine cell markers (amylase and P48) and endocrine cell markers (insulin 1, insulin 2, and Ngn3). They also expressed, at a lower level, PDX-1, Isl-1, and the islet hormones pancreatic polypeptide, glucagon and somatostatin as well as GLUT2, the stem cell marker ABCG2 and PECAM, a marker of endothelial cells. It was further shown by immunocytochemistry of the E15.5 pancreas that EGFP colocalised in separate subpopulations of cells that expressed nestin, insulin and amylase. These results support the conclusion that nestin expressing cells can give rise to both endocrine and exocrine cells. The ability to purify these putative progenitor cells may provide further insights into their properties and function. PMID- 16698178 TI - Recovery of nickel and cobalt from organic acid complexes: adsorption mechanisms of metal-organic complexes onto aminophosphonate chelating resin. AB - This study examined the recovery of nickel and cobalt from organic acid complexes using a chelating aminophosphonate Purolite S950 resin. These metal complexes are generated by bioleaching nickel laterite ores, a commercial nickel and cobalt mineral oxide, with heterotrophic organism and their metabolites or organic acid products. Equilibrium adsorption tests were conducted as a function of Ni and Co concentrations (15-2000 mg/L), solution pH (0.01 and 0.1 M acids) and three metabolic complexing agents (citrate, malate and lactate). It was shown that the adsorption of the various Ni- and Co-complexes on Purolite were quite low, 16-18 and 5.4-9 mg/g of resin, respectively, in comparison to the smaller nickel ions and nickel sulfate. This was attributed to the bulky organic ligands which promoted crowding effect or steric hindrance. The adsorption of these complexes was further hampered by the strong affinity of the resin to H+ ions under acidic conditions. Mechanisms of adsorption, as inferred from the fitted empirical Langmuir and Freundlich models, were correlated to the proposed steric hindrance and competitive adsorption effects. Nickel and cobalt elution from the resin were found be effective and were independent of the type of metal complexes and metal concentrations. This study demonstrated the relative challenges involved in recovering nickel and cobalt from bioleaching solutions. PMID- 16698179 TI - Anomalous sensorimotor integration in adults who stutter: a tendon vibration study. AB - Anomalies in oral movement control have been identified in stuttering, which suggest this speech disorder involves a sensorimotor deficit. To test whether adults who stutter (AWS) display aberrant proprioceptive function, masseter tendon vibration was used to manipulate jaw proprioception as AWS and normal speakers performed a jaw-opening task. A movement amplitude reduction in the vibration condition was observed in both groups indicating the movements of AWS and controls were influenced in a similar manner by altering masseter proprioception. However, the undershoot magnitude was reduced in AWS relative to the control participants indicating a subtle difference in proprioceptive integration among the stuttering participants. Our interpretation is that AWS use proprioceptive information less efficiently than normal speakers, which could interfere with sensorimotor integration during speech production. PMID- 16698180 TI - Ischemia-induced increase in RGS7 mRNA expression in gerbil hippocampus. AB - The present study investigated the changes in the expression of regulators of G protein-coupled signaling proteins RGS2, 7 and 8 in gerbil hippocampus to better understand alterations of G-protein-coupled receptors signaling after cerebral ischemia. In situ hybridization revealed a transient, robust early increase in RGS7 mRNA levels in the dentate gyrus after ischemia. RGS8 mRNA expression started to increase at a later time point in the CA3 region but no changes were found for RGS2. Our results show a subtype-, time-, and subregion-specific regulation in mRNA expression of RGS proteins after cerebral ischemia in gerbil hippocampus. PMID- 16698181 TI - Psychiatric comorbidities in a community sample of women with fibromyalgia. AB - Prior studies of careseeking fibromyalgia (FM) patients often report that they have an elevated risk of psychiatric disorders, but biased sampling may distort true risk. The current investigation utilizes state-of-the-art diagnostic procedures for both FM and psychiatric disorders to estimate prevalence rates of FM and the comorbidity of FM and specific psychiatric disorders in a diverse community sample of women. Participants were screened by telephone for FM and MDD, by randomly selecting telephone numbers from a list of households with women in the NY/NJ metropolitan area. Eligible women were invited to complete physical examinations for FM and clinician-administered psychiatric interviews. Data were weighted to adjust for sampling procedures and population demographics. The estimated overall prevalence of FM among women in the NY/NJ metropolitan area was 3.7% (95% CI=3.2, 4.4), with higher rates among racial minorities. Although risk of current MDD was nearly 3-fold higher in community women with than without FM, the groups had similar risk of lifetime MDD. Risk of lifetime anxiety disorders, particularly obsessive compulsive disorder and post-traumatic stress disorder, was approximately 5-fold higher among women with FM. Overall, this study found a community prevalence for FM among women that replicates prior North American studies, and revealed that FM may be even more prevalent among racial minority women. These community-based data also indicate that the relationship between MDD and FM may be more complicated than previously thought, and call for an increased focus on anxiety disorders in FM. PMID- 16698182 TI - Inactivation of O6-alkylguanine DNA alkyltransferase as a means to enhance chemotherapy. AB - DNA adducts at the O6-position of guanine are a result of the carcinogenic, mutagenic and cytotoxic actions of methylating and chloroethylating agents. The presence of the DNA repair protein O6-alkylguanine-DNA alkyltransferase (AGT) renders cells resistant to the biological effects induced by agents that attack at this position. O6-Benzylguanine (O6-BG) is a low molecular weight substrate of AGT and therefore, results in sensitizing cells and tumors to alkylating agent induced cytotoxicity and antitumor activity. Presently, chemotherapy regimens of O6-BG in combination with BCNU, temozolomide and Gliadel are in clinical development. Other ongoing clinical trials include expression of mutant AGT proteins that confer resistance to O6-BG in bone marrow stem cells, in an effort to reduce the potential enhanced toxicity and mutagenicity of alkylating agents in the bone marrow. O6-BG has also been found to enhance the cytotoxicity of agents that do not form adducts at the O6-position of DNA, including platinating agents. O6-BG's mechanism of action with these agents is not fully understood; however, it is independent of AGT activity or AGT inactivation. A better understanding of the effects of this agent will contribute to its clinical usefulness and the design of better analogs to further improve cancer chemotherapy. PMID- 16698183 TI - Insight into the treatment of cancer in older patients: developments in the last decade. AB - In the last decades there has been an increased interest in the treatment of elderly cancer patients and a change in attitude of both clinicians and their patients has occurred. Drugs are now available that might be considered "elderly friendly" and the enormous advances in surgical procedures and supportive treatments over the recent years have enabled adverse effects to be minimized. A Geriatric Assessment is increasingly used as a tool to define those patients who are more suitable for aggressive chemotherapy or, on the contrary, palliative treatment. For almost all cancers, older patients are better treated today than they were in the past, even though we are still far from optimal management. Despite the perceived barriers to including elderly patients in clinical trials, there are few data to support excluding them. We must not permit increased age in cancer patients to continue to be an important and independent risk factor for receiving inadequate care. PMID- 16698184 TI - A comparison of ventral tegmental neurons projecting to optic flow regions of the inferior olive vs. the hippocampal formation. AB - The ventral tegmental area (catecholaminergic group A10) is a midbrain region characterized by concentrated dopaminergic immunoreactivity. Previous studies in pigeons show that the ventral tegmental area provides a robust projection to the hippocampal formation and to the medial column of the inferior olive. However, the distribution, morphology, and neurochemical content of the neurons that constitute these projections have not been resolved. In this study, we used a combination of retrograde tracing techniques and immunofluorohistochemistry to address these issues. Retrograde tracers were used to demonstrate that the distribution of ventral tegmental area neurons projecting to the hippocampus and the inferior olive overlap in the caudo-ventral ventral tegmental area. The hippocampus- and inferior olive-projecting ventral tegmental area neurons could not be distinguished based on morphology: most neurons had small- to medium-sized multipolar or fusiform soma. Double-labeling with fluorescent retrograde tracers revealed that the hippocampus- and medial column of the inferior olive-projecting neurons were found intermingled in the ventral tegmental area, but no cells were double labeled; i.e. individual ventral tegmental area neurons do not project to both the hippocampal formation and medial column of the inferior olive. Finally, we found that a minority (8.2%) of ventral tegmental area neurons providing input to the hippocampus were tyrosine hydroxylase-immunoreactive, whereas none of the inferior olive-projecting neurons were tyrosine hydroxylase positive. Combined, our findings show that the projections to the hippocampus and olivocerebellar pathway arise from intermixed subpopulations of ventral tegmental area neurons with indistinguishable morphology but only the hippocampal projection involves dopaminergic neurons. We suggest that equivalent projections from the ventral tegmental area to the hippocampal formation and inferior olive exist in mammals and discuss their potential role in the processing of optic flow and the analysis of self-motion. PMID- 16698185 TI - A longitudinal study on cortisol and complaint reduction in burnout. AB - Several studies have investigated the association between burnout and HPA-axis functioning, but the results are far from consistent. This does not preclude the possibility that within a group of burnout patients a recovery of symptoms in a longitudinal course corresponds to (changes in) cortisol parameters. The latter possibility is tested in the present study before and after treatment, and at follow-up. HPA-axis functioning and burnout complaints were assessed in burned out participants at baseline (n=74), post-treatment (n=62) and at follow-up (n=53). Multilevel regression analysis was used to test the hypothesis. Burnout complaints were significantly reduced at 8.5 months post-treatment, but there was no further reduction in complaints at follow-up 6.3 months later. Cortisol after awakening, and after dexamethasone intake showed no changes from baseline to post treatment and follow-up. There was a small decline in cortisol during the day over the longitudinal course. The cortisol level after awakening in the longitudinal course showed significant positive association with the initial exhaustion level, a negative association with the change in the burnout exhaustion score, and a positive association with the change in depression. Although these associations are statistically significant, they only explain a small fraction of the variance in cortisol after awakening between and within persons. This implies that changes at symptom level are hardly related to changes in cortisol functioning, therefore the clinical implications of this finding are limited. PMID- 16698187 TI - The anterior cruciate ligament deficiency as a model of brain plasticity. AB - This paper describes a plausible mechanism for the development of brain plastic changes due to a peripheral joint injury, such as anterior cruciate ligament (ACL) rupture. Evidence for the hypothesis is based on mainly three indications derived from the literature review: (a) the existence of two different categories of ACL patients, the copers and non-copers, presenting different features of functional deficiencies, (b) the demand of a sufficient post-traumatic time (more than 6 months) for the dysfunction development and (c) the fact that the dysfunction is not limited to the injured limb but also concerns the non-injured one. Considering the fact that ACL contains mechanoreceptors, which inform the central nervous system (CNS) about joint sense position and kinaesthesia, it can be suggested that this kind of injury might be regarded as a neurophysiological dysfunction, not being a simple musculoskeletal injury. The rupture of the ACL could lead to the cessation/depletion or differentiation of the ascending afferent pathway from the injured joint towards CNS, inducing to the joint de afferentation and consequently CNS reorganization and joint de-efferentation. In case of presumable evidence of the proposed hypothesis, its clinical application could concern several aspects of the intervention procedures. For example, a number of clinical findings, such as the functional differences presented between two separate clinical groups of patients (copers and non-copers) could be justified or the rehabilitation strategies might have to be revised, provided that certain therapeutic components have influence on facilitating brain plastic changes that induce to beneficial functional outcomes. PMID- 16698186 TI - Increased coronary flow may prolong transmural ventricular repolarisation. AB - An increase in spatial dispersion of ventricular refractory periods reflects inhomogeneity of ventricular refractoriness and is associated with an increased risk of life-threatening ventricular arrhythmias. Spatial dispersion in ventricular refractoriness is determined by the differences in refractory periods of myocytes in epicardium, mid-myocardium and endocardium. Prolongation of ventricular repolarisation with drugs or physical means may reduce the spatial dispersion and the risk of arrhythmia. An increase in coronary flow has been shown to prolong the duration of ventricular repolarisation measured from epicardium in the intact animal heart. We hypothesised that an increase in coronary flow may also prolong ventricular repolarisation cross the three layers of ventricular myocardium through the release of nitric oxide. The simultaneously prolongation of transmural repolarisation may reduce the spatial dispersion of ventricular repolarisation and hence, the risk of life-threatening ventricular arrhythmias. PMID- 16698188 TI - Fatalities after taking ibogaine in addiction treatment could be related to sudden cardiac death caused by autonomic dysfunction. AB - Ibogaine is the most important alkaloid of the Central African Iboga-shrub. It is the central drug in Gabonian initiation ceremonies in which it is used to cause a near-death experience. In Western countries it is used in private clinics to treat addiction. However, in the United States and most European countries it is classified as an illegal drug because at least eight persons have died after having taken Ibogaine. These fatalities occurred in most cases several days after ingestion or following the intake of very small doses. There is no conclusive explanation at the present time for these deaths. We hypothesize, that these deaths may be a result of cardiac arrhythmias, caused by a dysregulation of the autonomic nervous system. Ibogaine affects the autonomic nervous system by influencing several neurotransmitter-systems and the fastigial nucleus. The cerebellar nucleus responds to small doses with a stimulation of the sympathetic system, leading to a fight or flight reaction. High doses, however, lead to a vagal dominance: a "feigned death". The risk of cardiac arrhythmias is increased in situations of sympathetic stimulation or coincidence of a high parasympathetic tonus and a left-sided sympathetic stimulation. This could occur under influence of small doses of ibogaine and also at times of exhaustion with a high vagal tonus, when sudden fear reactions could cause a critical left-sided sympathetic stimulation. Gabonian healers prevent these risks by isolating their patients from normal life and by inducing a trance-state with right-hemispheric and vagal dominance for several days. PMID- 16698189 TI - Brain profiling and clinical-neuroscience. AB - The current psychiatric diagnostic system, the diagnostic statistic manual, has recently come under increasing criticism. The major reason for the shortcomings of the current psychiatric diagnosis is the lack of a scientific brain-related etiological knowledge about mental disorders. The advancement toward such knowledge is further hampered by the lack of a theoretical framework or "language" that translates clinical findings of mental disorders to brain disturbances and insufficiencies. Here such a theoretical construct is proposed based on insights from neuroscience and neural-computation models. Correlates between clinical manifestations and presumed neuronal network disturbances are proposed in the form of a practical diagnostic system titled "Brain Profiling". Three dimensions make-up brain profiling, "neural complexity disorders", "neuronal resilience insufficiency", and "context-sensitive processing decline". The first dimension relates to disturbances occurring to fast neuronal activations in the millisecond range, it incorporates connectivity and hierarchical imbalances appertaining typically to psychotic and schizophrenic clinical manifestations. The second dimension relates to disturbances that alter slower changes namely long-term synaptic modulations, and incorporates disturbances to optimization and constraint satisfactions within relevant neuronal circuitry. Finally, the level of internal representations related to personality disorders is presented by a "context-sensitive process decline" as the third dimension. For practical use of brain profiling diagnosis a consensual list of psychiatric clinical manifestations provides a "diagnostic input vector", clinical findings are coded 1 for "detection" and 0 for "non-detection", 0.5 is coded for "questionable". The entries are clustered according to their presumed neuronal dynamic relationships and coefficients determine their relevance to the specific related brain disturbance. Relevant equations calculate and normalize the different values attributed to relevant brain disturbances culminating in a three-digit estimation representing the three diagnostic dimensions. brain profiling has the promise for a future brain-related diagnosis. It offers testable predictions about the etiology of mental disorders because being brain related it lends readily to brain imaging investigations. Being presented also as a one-point representation in a three-dimensional space, multiple follow-up diagnoses trace a trajectory representing an easy-to-see clinical history of the patient. Additional, more immediate, advantages involve reduced stigma because it relaters the disorder to the brain not the person, in addition the three-digit diagnostic code is clinically informative unlike the DSM codes that have no clinical relevance. To conclude, brain profiling diagnosis of mental disorders could be a bold new step toward a "clinical-neuroscience" substituting "psychiatry". PMID- 16698190 TI - Triglycerides from Urena lobata. AB - Two triglycerides, both bearing polyunsaturated fatty acid residues, were isolated from the hexane extract of Urena lobata. One of them is characterized by the presence of three different polyunsaturated fatty acids. Their structures were studied with spectral methods. PMID- 16698191 TI - Wound healing potential of Tragia involucrata extract in rats. AB - The methanol extract of the roots of Tragia involucrata topically tested at doses of 100 and 200 mg/kg exerted significant wound healing effect in Staphylococcus aureus-induced excision wound in rats. PMID- 16698192 TI - Antioxidant activity of Punica granatum fruits. AB - Arils, juice and rinds of Punica granatum fruits and their aqueous and ethyl acetate extracts displayed good antioxidant activity. PMID- 16698193 TI - A new cerebroside from Polygala japonica. AB - The isolation of a new cerebroside, 1-O-beta-d-glucopyranosyl-(2S,3S,4R,11E)-2 [(2'R)-2'-hydroxypalmitoylamino]-11-octadecene-1,3,4-triol (polygalacerebroside) (1), from the aerial part of Polygala japonica is reported. PMID- 16698194 TI - Sedative activity of stem bark of the Sri Lankan endemic plant, Vateria copallifera. AB - An aqueous bark extract of Vateria copallifera showed marked sedative activity, which supports the therapeutic effects claimed by Sri Lankan traditional physicians. PMID- 16698196 TI - [Biological diagnosis of systemic candidiasis: difficulties and future prospects]. AB - The diagnosis of systemic Candidiasis is difficult to establish and biologic diagnosis raises problems. Blood culture which is the gold standard for the diagnosis of systemic Candidiasis lacks sensitivity and usually takes several days to become positive. Early diagnostic approach is imperative to avoid delays in the initiation for treatment. Therefore, nonculture methods like test for Candida antigen detection, metabolite detection or Candida DNA detection by PCR are being developed for the laboratory diagnosis. Candida derived metabolites and antigens detection lacks sensitivity. A new strategy consisting of the combined detection of mannanemia and an antibody response was developed. The combined detection has a high specificity and sensitivity in the diagnosis of invasive candidiasis. The results of tests for the detection of yeast DNA by PCR obtained recently are promising in terms of sensitivity, specificity and identification of species of Candida. PMID- 16698195 TI - COX-2 and sPLA2 inhibitory activity of aqueous extract and polyphenols of Rhizophora mangle (red mangrove). AB - The aqueous extract of Rhizophora mangle bark and its polyphenolic fractions showed remarkable in vitro antiinflammatory activity in a preliminary study. The low molecular weight fraction exhibited cyclooxygenase-2 inhibitory activity while the total aqueous extract and the low molecular weight fraction showed secretory phospholipase A(2) inhibitory activity. PMID- 16698197 TI - Identification and characterization of the gene encoding the Acidobacterium capsulatum major sigma factor. AB - Acidobacterium capsulatum is an acid-tolerant, encapsulated, Gram-negative member of the ubiquitous, but poorly understood Acidobacteria phylum. Little is known about the genetics and regulatory mechanisms of A. capsulatum. To begin to address this gap, we identified the gene encoding the A. capsulatum major sigma factor, rpoD, which encodes a 597-amino acid protein with a predicted sequence highly similar to the major sigma factors of Solibacter usitatus Ellin6076 and Geobacter sulfurreducens PCA. Purified hexahistidine-tagged RpoD migrates at approximately 70 kDa under SDS-PAGE conditions, which is consistent with the predicted MW of 69.2 kDa, and the gene product is immunoreactive with monoclonal antibodies specific for either bacterial RpoD proteins or the N-terminal histidine tag. A. capsulatum RpoD restored normal growth to E. coli strain CAG20153 under conditions that prevent expression of the endogenous rpoD. These results indicate we have cloned the gene encoding the A. capsulatum major sigma factor and the gene product is active in E. coli. PMID- 16698198 TI - Vaccine use and disease prevalence in dogs and cats. AB - A yearly revaccination of adult pets against distemper, the adenoviral and parvoviral diseases is scientifically unwarranted, professionally obsolete and ethically questionable; other vaccinal antigens, however, may need yearly or even more frequent injections. Base immunisation is redefined: it is complete only after the multivalent booster in the second year of life. A yearly revaccination interview, not necessarily an injection, should become the new standard. This interview is a professional service that must be taught, expertly performed and invoiced. Adult animals should be "vaccinated to measure", taking age, breed, lifestyle, the epidemiologic situation, etc. into account. Post-vaccination serology should become a guide in revaccination decisions. For a solid herd immunity, more animals of the population must be vaccinated. The profession should issue regular updates of the 'code of vaccination practice'. To counteract vaccination antagonism, a concerted action of academia, the veterinary profession and industry is required. PMID- 16698199 TI - The challenge for the next generation of feline calicivirus vaccines. AB - Feline calicivirus (FCV) has been shown to evolve within individual cats and in the environment of colonies. This evolution and the diversity it creates has important clinical implications, not only for the disease in cats, but also for attempts to control disease by vaccination. Generally speaking, existing vaccines appear to be very effective at controlling the majority of clinical disease. However, some concerns remain including a failure to induce sterilising immunity, occasional vaccine breakdowns, and for live vaccines, occasional vaccine-induced disease. Key areas for future vaccine development include monitoring and broadening the cross-reactivity of vaccine immunity to field viruses, especially the recently evolved highly virulent strains, and attempting to reduce/eliminate field virus shedding by vaccinated cats. PMID- 16698200 TI - Comparison of IS900 tissue PCR, bacterial culture, johnin and serological tests for diagnosis of naturally occurring paratuberculosis in goats. AB - Comparative efficacy of an IS900 tissue PCR, bacterial culture, johnin, agar-gel immunodiffusion (AGID) and absorbed-ELISA tests was investigated in 43 goats naturally infected with paratuberculosis. On histological examination, tissue sections from all animals showed typical granulomatous inflammatory changes. The lesions were classified as multibacillary (MB) (n=30), which had diffuse granulomatous lesions with abundant acid-fast bacilli (AFB), and paucibacillary (PB) (n=13), which had focal or multifocal granulomatous lesions with few AFB. The sensitivities of johnin test, tissue culture, faecal culture, tissue PCR, AGID and ELISA were 68% (17/25), 100% (30/30), 84.6% (22/26), 100% (30/30), 96.2% (25/26) and 100% (26/26) in MB goats, and 88.8 (8/9), 46.1% (8/13), 40% (4/10), 61.5% (8/13), 50% (5/10), and 70% (7/10) in PB goats, respectively. Except for the johnin test, which showed higher sensitivity in PB goats, all other tests displayed significantly higher sensitivities in MB goats. The results indicate the usefulness of tissue PCR, culture and serological tests in the diagnosis of clinically affected paratuberculous goats, especially with multibacillary pathology. PMID- 16698201 TI - Serological testing--an alternative to boosters? AB - The issue of the duration of immunity, particularly for the modified live viral components of veterinary vaccines, has been a significant part of the recent vaccination debate. One manufacturer has increased the recommended booster interval for these components to 3 years give name and another now states 'up to 4 years' immunity. There remain many unanswered questions regarding this duration of immunity (DOI). Studies suitable for data sheet claims are time consuming and costly and can only be performed in laboratory dogs under tightly controlled conditions. Evidence from rabies serology testing in the UK shows that the response of individual animals to routine vaccination is highly variable. Much of the published field evidence on the persistence of antibody titres originates from North America, where vaccination strategies and reservoir species differ from Europe. Quantifying the effect of exposure to field virus on the maintenance of immunity in these studies is impossible, and little is known of the circulation of virus in unvaccinated dogs and wild mammals throughout Europe. If owners or vets are concerned about revaccination one option is to assess the need for each booster by performing a blood test. There is some published evidence of the relationship between antibody titres and protective immunity, and tests are available to measure responses to individual viral components of the routine canine and feline vaccines. It must be remembered that most commercial tests to assess immunity only measure antibodies, which are only one aspect of the immune response to vaccination. It is therefore possible that animals without or with low antibody titres are in fact protected. Serological tests are an option if owners are unwilling to have their animal boostered without evidence that it is needed. However, the cost of these tests is likely to exceed that of booster vaccination for the foreseeable future. PMID- 16698202 TI - Cryopreservation of in vitro matured buffalo (Bubalus bubalis) oocytes by minimum volumes vitrification methods. AB - The aim of this study was to evaluate the efficiency of the solid surface vitrification (SSV) and the cryoloop vitrification (CLV) methods to cryopreserve in vitro matured buffalo oocytes. Another objective of the work was to investigate whether the presence of cumulus cells affects the efficiency of oocyte vitrification in this species. In the SSV method, oocytes were vitrified in a solution of 35% ethylene glycol, 5% polyvinyl-pyrrolidone and 0.4% trehalose and they were warmed in a 0.3M trehalose solution. In the CLV method, oocytes were vitrified in 16.5% ethylene glycol and 16.5% dimethyl sulfoxide and warmed in decreasing concentrations of sucrose. The oocytes that survived vitrification were fertilized and cultured in vitro up to the blastocyst stage. Although high survival rates were recorded in all groups, when the oocytes were vitrified by the CLV method in the absence of cumulus cells, the survival rate was significantly (P<0.05) lower. However, the CLV gave a significantly higher cleavage rate compared to the SSV with the denuded oocytes (45% versus 26%, respectively; P<0.05), whereas no differences were found between methods with the cumulus-enclosed oocytes (14% versus 15%, respectively). Blastocysts were produced for the first time from in vitro matured oocytes that were vitrified warmed in buffalo. Nevertheless, vitrification significantly decreased blastocyst yield, regardless of both the method employed and the presence or absence of cumulus cells. PMID- 16698203 TI - Characterizing powder mixing processes utilizing compartment models. AB - Powder mixing has been the subject of substantial research due to its importance in a variety of industrial sectors, including pharmaceuticals, food, and polymer manufacturing. Although a number of different models have been proposed in the literature, most of them are either empirical or require computationally intensive calculations that make them difficult to implement for realistic systems. The aim of this paper is to develop a simplified framework, based on compartment modeling that efficiently and accurately captures the system behavior. Using the V-blender as a model system, the compartment modeling approach was used to illustrate the effects of vessel loading on mixing as well as the impact of sampling methods on the accuracy of mixing characterization. PMID- 16698204 TI - QSPR models for the prediction of apparent volume of distribution. AB - An estimate of volume of distribution (V(d)) is of paramount importance both in drug choice as well as maintenance and loading dose calculations in therapeutics. It can also be used in the prediction of drug biological half life. This study employs quantitative structure-pharmacokinetic relationship (QSPR) techniques for the prediction of volume of distribution. Values of V(d) for 129 drugs were collated from the literature. Structural descriptors consisted of partitioning, quantum mechanical, molecular mechanical, and connectivity parameters calculated by specialized software and pK(a) values obtained from ACD labs/log D database. Genetic algorithm and stepwise regression analyses were used for variable selection and model development. Models were validated using a leave-many-out procedure. QSPR analyses resulted in a number of significant models for acidic and basic drugs separately, and for all the drugs. Validation studies showed that mean fold error of predictions for the selected models were between 1.79 and 2.17. Although separate QSPR models for acids and bases resulted in lower prediction errors than models for all the drugs, the external validation study showed a limited applicability for the equation obtained for acids. Therefore, the universal model that requires only calculated structural descriptors was recommended. The QSPR model is able to predict the volume of distribution of drugs belonging to different chemical classes with a prediction error similar to that of the other more complicated prediction methods including the commonly practiced interspecies scaling. The structural descriptors in the model can be interpreted based on the known mechanisms of distribution and the molecular structures of the drugs. PMID- 16698205 TI - The spermatogenic and ovogenic effects of chronically administered Shilajit to rats. AB - This study examined the possibility of using Shilajit as a fertility agent. The effects of Shilajit on spermatogenesis and ovogenesis were studied using male and female rats. Shilajit was administered orally to 7-week-old rats over a 6-week period. In the male rats, the number of sperms in the testes and epididymides was significant higher than in the control. A histological examination revealed an apparent increase in the number of seminiferous tubular cell layers in the testes of the treated rats. However, there were no significant differences in the weights of heart, spleen, liver, kidney, brain, testes and epididymides. In the female rats, the effect of Shilajit was estimated by the ovulation inducing activity. Over a 5-day, ovulation was induced in seven out of nine rats in the Shilajit administration group and in three out of nine rats in the control. It was estimated that Shilajit had both a spermiogenic and ovogenic effect in mature rats. PMID- 16698206 TI - Effect of Mouriri pusa extracts on experimentally induced gastric lesions in rodents: role of endogenous sulfhydryls compounds and nitric oxide in gastroprotection. AB - Several plants are used in folk medicine to treat gastrointestinal disorders. Mouriri pusa Gardn. (Melastomataceae) is a medicinal plant commonly used in the central region of Brazil against gastric ulcer. Two organic extracts methanolic (MeOH) and dichloromethane (DCM) obtained by sequential extraction from the leaves of Mouriri pusa were evaluated for their ability to protect the gastric mucosa against injuries caused by necrotizing agents (0.3M HCl/60% EtOH, absolute ethanol, non-steroidal anti-inflammatory drug, stress and pylorus ligature) in mice and rats. The best results were obtained after pretreatment with MeOH extract whereas the DCM extract did not show the same significant antiulcerogenic activity. No acute toxicity was observed in animals treated with 5 g/kg, p.o. of MeOH extract. The mechanism involving the antiulcerogenic action of MeOH extract seemed to be related to NO generation and also suggested the effective participation of endogenous sulfhydryl group in the gastroprotective action. Phytochemical investigation of the MeOH extract of Mouriri pusa yielded tannins, flavonoids and (-)-epicatechin. The presence of these phenolic compounds probably would explain the antiulcerogenic effect of the polar extract of Mouriri pusa leaves. PMID- 16698208 TI - Anti-infective potential of natural products: how to develop a stronger in vitro 'proof-of-concept'. AB - Natural products, either as pure compounds or as standardized plant extracts, provide unlimited opportunities for new drug leads because of the unmatched availability of chemical diversity. To secure this, a number of pivotal quality standards need to be set at the level of extract processing and primary evaluation in pharmacological screening models. This review provides a number of recommendations that will help to define a more sound 'proof-of-concept' for antibacterial, antifungal, antiviral and antiparasitic potential in natural products. An integrated panel of pathogens is proposed for antimicrobial profiling, using accessible standard in vitro experimental procedures, endpoint parameters and efficacy criteria. Primary requirements include: (1) use of reference strains or fully characterized clinical isolates, (2) in vitro models on the whole organism and if possible cell-based, (3) evaluation of selectivity by parallel cytotoxicity testing and/or integrated profiling against unrelated micro-organisms, (4) adequately broad dose range, enabling dose-response curves, (5) stringent endpoint criteria with IC(50)-values generally below 100microug/ml for extracts and below 25microM for pure compounds, (6) proper preparation, storage and in-test processing of extracts, (7) inclusion of appropriate controls in each in vitro test replicate (blanks, infected and reference controls) and (8) follow-up of in vitro activity ('hit'-status) in matching animal models ('lead' status). PMID- 16698207 TI - Protective effect of Rehmannia glutinosa on the cisplatin-induced damage of HEI OC1 auditory cells through scavenging free radicals. AB - The steamed root of Rehmannia glutinosa has been used in traditional Oriental Medicine for treatment of inner ear diseases, such as tinnitus and hearing loss. In the present study, we showed that the ethanol extract of steamed roots of Rehmannia glutinosa (SRG) protected HEI-OC1 auditory cells from cisplatin cytotoxicity in a dose-dependent fashion. In addition, to investigate the protection mechanism of SRG on cisplatin cytotoxicity towards HEI-OC1, we measured the effects of SRG on lipid peroxidation of cisplatin treated cells as well as scavenging activities against superoxide radical, hydroxyl radical, hydrogen peroxide, and DPPH radical. SRG (5-100 microg/ml) had protective effect against the cisplatin-induced HEI-OC1 cell damage and reduced lipid peroxidation in a dose-dependent manner. Furthermore, SRG showed strong scavenging activity against superoxide radical, hydroxyl radical, hydrogen peroxide, and DPPH radical. These results indicate that SRG protects cisplatin-induced HEI-OC1 cell damage through inhibition of lipid peroxidation and scavenging activities of free radials. PMID- 16698209 TI - [Epidemiology, control and treatments of antimicrobial resistances: highlights of the 45th ICAAC, Washington, 2005]. AB - For more than 20 years, hospital and community-acquired antimicrobial resistances regularly increase. In France, methicilline-resistant Staphylococcus aureus (MRSA) and extended-spectrum beta-lactamase-producing Enterobacteriaceae are the most common resistant pathogens. Previous hospitalisation, surgery, long length of stay, enteral feeding and antibiotic exposure are the main risk factors associated with nosocomial MRSA infections. To limit the emergence of resistances, control strategies have been implemented, based on isolation practices, healthcare workers education, strict hand hygiene and a controlled use of antibiotics. However, new antimicrobials acting by a novel mechanism of action are necessary in fighting the most resistant organisms. Therefore, the launch on the American market in June 2005 of tigecycline, a first-in-class glycylcycline indicated in the treatment of complicated skin and skin structure infections and complicated intra-abdominal infections, will provide to physicians a new therapeutic option against hospital or community-acquired resistant pathogens. The data presented to ICAAC indicate its expended broad spectrum of in vitro activity against Gram positives, Gram negatives, anaerobes and against number of difficult-to-treat resistant isolates, such as hospital or community-acquired MRSA, vancomycine resistant Enterococcus but also ESBL-producing organisms. PMID- 16698210 TI - [Antibiotic therapy in serious infectious states]. AB - Severe sepsis, which is related to a high mortality rate, requires a very specific antibiotic strategy, which must be adapted to each case. The appropriateness of empiric therapy is based on the delay before administration of the molecule, the bacterial resistance profile, and the kinetic and/or dynamic properties of the available antibiotics. PMID- 16698211 TI - CD34+/CD133- circulating endothelial precursor cells (CEP): characterization, senescence and in vivo application. AB - Circulating endothelial precursor cells (CEP) are interesting candidates for the treatment of ischemic diseases and for tumor targeting/imaging. We isolated a homogeneous population of CEP from CD34(+)/CD133(-) cells of peripheral blood that can be expanded easily on collagen-type-I coated plastic. CEP displayed a phenotype of mature endothelial cells (vWF, CD31, CD34, VEGF-R2, CD105, CD146) similar to that of cord-blood CEP and umbilical vein endothelial cells. They bound UEA-1 lectin, incorporated acetylated LDL and formed tube-like structures with capillary lumens in vitro. Weibel-Palade bodies were observed by electron microscopy. After 40-60 cell population doublings, CEP cultures underwent a terminal growth arrest, had shorter telomeres, up-regulated cell cycle inhibitory proteins, such as p21(CIP1) and stained positive for senescence-associated-beta galactosidase. During the whole expansion period CEP retained their endothelial phenotype and a normal karyotype. CEP had the capacity to home to ischemic tissue in vivo after systemic injection in nude rats. The convenient expandability, the homogenous phenotype, the functional cellular senescence program, the regular karyotype and the homing capacity to ischemic myocardium suggest autologous CEP cultures as a safe and promising tool for cell-based therapeutic approaches in targeting ischemic tissue and tumors. PMID- 16698212 TI - A rapid and simple procedure for the determination of synephrine in dietary supplements by gas chromatography-mass spectrometry. AB - A simple and rapid procedure based on gas chromatography-mass spectrometry (GC MS) is described for determination of synephrine, active principle of Citrus aurantium plant, in solid and liquid dietary supplements. After the addition of 3,4-methylenedioxypropylamphetamine as internal standard (I.S.), a liquid-liquid extraction procedure in alkaline conditions with chloroform/isopropanol (9:1, v/v) was applied to the samples prior to analysis. Chromatography was performed on a fused capillary column and synephrine and I.S., derivatized with pentafluoropropionic anhydride, were determined in the selected-ion-monitoring (SIM) mode. The method was validated in the range 0.1-50 microg/mg or microg/mL synephrine. Mean recovery ranged between 89.3% and 90.5% in both solid and liquid dietary supplements. The quantification limit was 0.1 microg/mg or microg/ml. The method was applied to analysis of various dietary supplements promoted for aiding weight control containing, among other constituents such as ephedrine alkaloids and methylxanthines, Citrus aurantium. Amount of synephrine present in such products ranged from 3.1 microg/mg solid product to 480.2 microg/mL liquid product. PMID- 16698213 TI - In vivo and real time determination of ornidazole and tinidazole and pharmacokinetic study by capillary electrophoresis with microdialysis. AB - The aim of this study was to develop a rapid and sensitive method for in vivo and real time monitoring unbound ornidazole (ONZ) and tinidazole (TNZ) in rabbit blood using capillary electrophoresis coupled with microdialysis. The UV wavelength was set at 214 nm and all separations were performed in 20 mM Tris H3PO4 (pH 1.5) buffer. Microdialysis probes were perfused at 4 microl/min resulting in relative recoveries of 33.1+/-3.6% and 34.8+/-3.3% (n=3) for ONZ and TNZ, respectively. The linearity was studied in the concentration range of 1.0 412 microg/ml for ONZ and 1.0-520 microg/ml for TNZ. The detection limits were 0.7 microg/ml for ONZ and 0.6 microg/ml for TNZ (S/N=3). All separation could be achieved within 15 min. This method has been successfully applied to the pharmacokinetic study of ONZ and TNZ in rabbit blood. PMID- 16698214 TI - Evaluation of the Cepheid herpes simplex virus typing real-time PCR assay using dermal and genital specimens. AB - The Cepheid herpes simplex virus (HSV) (Cepheid, Sunnyvale, CA) typing multiplex real-time polymerase chain reaction (PCR) assay was evaluated for its ability to detect HSV in dermal and genital specimens stored in M5 media. Swab specimens (n = 114) for HSV testing were placed in M5 media and split between our laboratory and a highly experienced reference laboratory. Aliquots for testing with the Cepheid assay were processed using a simple boil-and-go procedure and then run in a SmartCycler II (Cepheid). Aliquots tested at the reference laboratory were processed using a MagNA Pure LC DNA extractor (Roche Molecular Systems, Alameda, CA) and tested by the Roche HSV real-time PCR assay. Both laboratories detected 35 positives. Of the positive specimens, the Cepheid assay typed 16 as HSV 1 and 19 as HSV 2; the reference laboratory typed 15 as HSV 1, 19 as HSV 2, and 1 as HSV indeterminate. Our results demonstrate that the Cepheid real-time PCR assay, using specimens subjected to minimal specimen processing, performed as well as the Roche real-time PCR assay, using DNA extracts, for the detection of HSV DNA in genital and dermal specimens. PMID- 16698216 TI - Flavophospholipol use in animals: positive implications for antimicrobial resistance based on its microbiologic properties. AB - Bambermycin (flavophospholipol) is a phosphoglycolipid antimicrobial produced by various strains of Streptomyces. It is active primarily against Gram-positive bacteria because of inhibition of transglycosylase and thus of cell wall synthesis. Bambermycin is used as a feed additive growth promoter in cattle, pigs, chickens, and turkeys, but has no therapeutic use in humans or animals. Flavophospholipol is known to suppress certain microorganisms (e.g., Staphylococcus spp. and Enterococcus faecalis) and thus contributes to an improved equilibrium of the gut microflora providing a barrier to colonization with pathogenic bacteria and resultant improved weight gain and feed conversion. Flavophospholipol has also been shown to decrease the frequency of transferable drug resistance among Gram-negative enteropathogens and to reduce the shedding of pathogenic bacteria such as Salmonella in pigs, calves, and chickens. Plasmid mediated resistance to bambermycin has not been described. Likewise, cross resistance among bacteria between bambermycin and penicillin, tetracycline, streptomycin, erythromycin, or oleandromycin has not been observed. This brief review summarizes the antimicrobial properties of bambermycin, in particular, its potentially favorable role in decreasing antimicrobial resistance. PMID- 16698215 TI - The association between anatomic site of Candida colonization, invasive candidiasis, and mortality in critically ill surgical patients. AB - We evaluated whether the likelihood of developing invasive candidiasis (IC) differed depending upon the anatomic site of Candida colonization in 182 surgical intensive care unit (SICU) patients who participated in a randomized trial of fluconazole to prevent candidiasis. We also determined the impact of Candida colonization of different anatomic sites on all-cause SICU and hospital mortality. A total of 2851 surveillance fungal cultures collected from 5 anatomic sites were analyzed. There was a statistically significant difference in the frequency of IC comparing patients with and without urinary (13.2% versus 2.8%, P = .02), respiratory (8.0% versus 1.2%, P = .04), and rectum/ostomy (8.4% versus 0%, P = .01) colonization. Patients with negative rectum/ostomy cultures and patients with both negative urine and respiratory tract cultures did not develop IC. Candiduria detected at any time in the SICU was independently associated with SICU mortality (odds ratio, 2.86; 95% confidence interval, 1.05-7.74). Surveillance fungal cultures of particular anatomic sites may help differentiate patients at higher risk of developing IC from those at low risk. PMID- 16698217 TI - The suitable restriction enzymes for pulsed-field gel electrophoresis analysis of Bordetella pertussis. AB - We searched the restriction enzymes with rare cutting sites on the genome of Bordetella pertussis strain Tohama I using the Restriction Digest Tool software provided in the Institute for Genomic Research web site. The usefulness of 5 enzymes for pulsed-field gel electrophoresis (PFGE) analysis was evaluated with 68 B. pertussis isolates. The results indicated that AflII, DraI, SpeI, and XbaI were useful enzymes, and AflII was the best one for PFGE analysis of B. pertussis isolates. PMID- 16698218 TI - Blood and urine samples as useful sources for the direct detection of tuberculosis by polymerase chain reaction. AB - The aim of the study was to assess the utility of the polymerase chain reaction (PCR) assay in blood and urine for the diagnosis of tuberculosis (TB). We prospectively evaluated the usefulness of PCR performed in blood and urine samples from patients with proved or probable TB compared with a control group of patients. The PCR technique was performed using IS6110 primers. We included in the study 57 patients (43 with definite TB and 14 with probable TB) and 26 controls. Blood and urine samples were drawn at the time of microbiologic diagnosis and 3, 6, 9, and 12 months later. Cultures were positive in the early period (<1 month after treatment) in 11 of 57 patients (19%) with probable or definite TB, in comparison with 42% of patients (24/57) who yielded a positive PCR (P = 0.02). Urine samples increased the sensitivity of PCR determination in blood samples by 10%. The PCR in blood and/or urine was positive in 41% of patients with pulmonary TB, in 36% of patients with extrapulmonary TB, and in 50% of patients with disseminated TB. Mycobacterium tuberculosis was still detectable by PCR in 5 of 13 patients with cured TB after 1 or more months of antituberculous treatment. The PCR detection of M. tuberculosis in blood and urine samples is useful for the diagnosis of different clinical forms of TB, mostly in those patients in which sample extraction is difficult or requires aggressive techniques. The sensitivity of this technique could be improved studying more than 1 sample in each patient, even after initiating an antituberculous treatment. PMID- 16698219 TI - Morphine exposure prevents up-regulation of MR and GR binding sites in the brain of adult male and female rats due to prenatal stress. AB - Our previous work demonstrated that the hormone response to stress and the negative feedback inhibition to these hormones are sex-dependently altered by prenatal morphine exposure in adult rats. An alteration in the glucocorticoid negative feedback inhibition is mediated by glucocorticoid receptors (GR) that are distributed throughout the brain, and mineralocorticoid receptors (MR) localized mainly in the hippocampus and involved in a tonic influence of brain functions. Therefore, the present study examined the binding characteristics of MR and GR in young adult male and female rats exposed prenatally (E11-E18) to morphine (10 mg/kg/2 x /day), saline or no treatment at all (controls). At 60-90 days of age, animals were adrenalectomized (ADX) 24 h prior to decapitation. The hippocampus and hypothalamus were dissected for saturation binding assays. The data demonstrate that prenatal stress due to maternal saline injections up regulates MR and GR binding in the hippocampus of adult male rats and this effect is prevented by prenatal morphine exposure. There is no effect of prenatal morphine exposure on GR binding in the hypothalamus of males. In female rats, prenatal morphine exposure does not affect the binding of MR and GR in the hippocampus or GR in the hypothalamus relative to controls; however, they are affected by ovarian hormone fluctuation. Moreover, prenatal stress decreases MR binding in the hippocampus of diestrous females and GR binding in the hypothalamus of estrous females. Both decreases are prevented by prenatal morphine exposure. Thus, the present study demonstrates that: (1) prenatal stress due to maternal saline injections alters MR and GR binding of adult male and female rats and is prevented by prenatal morphine exposure; (2) the MR and GR binding in adult female rats are affected by ovarian hormone fluctuations. PMID- 16698220 TI - Behavior ontogeny in the elevated plus-maze: prenatal stress effects. AB - Prenatal stress is a putative model for studying some psychopathological disorders. Indeed, submitting pregnant animals to stress leads to enhanced anxiety in the adult offspring. However, little is known about how prenatal stress effects interacts with anxiety throughout development. To study this issue, prenatally stressed rats were tested in the elevated plus-maze at different ages. During pregnancy female rats were submitted to uncontrollable electric foot shock sessions every other day or kept undisturbed (controls). After delivery, litters from control and stressed dams were left undisturbed from the 3rd to the 14th postnatal days. Male and female rats were tested in the elevated plus-maze at the ages of 30, 45 or 60 days. The following measures were taken in the elevated plus-maze: number of entries and time spent in the arms (or their extremities) and frequency and time spent in naturalistic behaviors (stretching, rearing, end exploring, grooming and head dipping). Decreases in the percentage of entries into and in the time spent (only females) in the open arms were shown by 60-day-old prenatally stressed rats, but not by 30- and 45-day old. Increased open arm ends exploration was shown by 45-day-old prenatally stressed males. Rearing behavior was found to increase with age, a phenomenon more pronounced in females. Additionally, at the younger ages prenatally stressed rats were heavier than controls, an effect which disappeared at young adulthood. In conclusion, anxiogenic prenatal stress effects in the elevated plus-maze could only be detected at early adulthood, not before. Nonetheless, at late adolescence (45 days of age) prenatal stress leaded to an anxiolytic-like effect which can be interpreted as increased risk-taking behavior. PMID- 16698221 TI - Effect of prostaglandin F2alpha on ovarian, adrenal, and pituitary hormones and on luteal blood flow in mares. AB - The effect of a single injection of prostaglandin F2alpha (PGF) during mid diestrus on systemic concentrations of progesterone, LH, FSH, estradiol, and cortisol and on blood flow to the corpus luteum was studied in 10 controls and 10 PGF-treated mares. Blood flow was assessed by estimating the percentage of corpus luteum with color-Doppler signals of blood flow during real-time scanning of the entire structure and by the diameter of the vascular pedicle near its attachment to the ovary. Treatment was done 8 days after ovulation and 0 h was immediately before the treatment. Examinations and collection of blood samples were done at 0 h, every 5 min until 1h, and then at 1.5, 2, 4, 8, 12, 24, 48, and 72 h. The concentrations of estradiol did not change, but progesterone, LH, FSH, and cortisol increased significantly within 5 min. Concentrations of LH and FSH in the PGF group remained elevated until a temporarily lower concentration at 8 or 4h, respectively, rebounded to 12h, and then slowly decreased. Cortisol remained elevated, until a decrease between 1 and 4h. Progesterone in the PGF group increased significantly until 10 min after 0 h and then decreased by 40 min to below the concentrations in controls. Within the PGF group, progesterone decreased significantly by 45 min to below the concentrations at 0 h. The values for each of the two indicators of blood flow did not differ significantly between the PGF and control groups until a decrease at 24h in the PGF group. Results did not support the hypothesis that the immediate transient post-PGF increase in progesterone was associated with an increase in luteal blood flow. Luteolysis, as indicated by decreasing progesterone, began well before the beginning of a decrease in luteal blood flow. PMID- 16698222 TI - Growth hormone suppresses the expression of IGFBP-5, and promotes the IGF-I induced phosphorylation of Akt in bovine mammary epithelial cells. AB - Growth hormone (GH) plays a specific role to inhibit apoptosis in the bovine mammary gland through the insulin-like growth factor (IGF)-I system, however, the mechanism of GH action is poorly understood. In this study, we show that GH dramatically inhibits the expression of IGFBP-5, and GH along with IGF-I enhanced the phosphorylation of Akt through the reduction of IGF binding protein (IGFBP) 5. To determine how GH affects Akt through IGF-I in bovine mammary epithelial cells (BMECs), we examined the phosphorylation of Akt in GH treated BMECs and found that IGF-I induced phosphorylation of Akt was significantly enhanced by the treatment with GH. We demonstrated that GH reduces mRNA and protein expression of IGFBP-5 in BMECs, but it does not affect the expression of IGFBP-3. To determine that the enhanced effect of the Akt phosphorylation by the treatment of GH is due to the inhibition of the expression of IGFBP-5, we examined the effect of IGFBP-3 and -5 on the phosphorylation of Akt through IGF-I in the GH-treated BMECs. The phosphorylation of Akt was inhibited in a dose-dependent manner when IGFBP-5 was added at varying concentrations and was also inhibited in the presence of IGFBP 3. The results of this study suggest that GH plays an important role on mammary gland involution in bovine mammary epithelial cells. PMID- 16698223 TI - Management of cervical cancer. AB - AIMS: The aim of the article was to review the current approach to management of cervical cancer. METHODS: The relevant literature has served as a source for review of different options applied in the management of cervical cancer. RESULTS: Treatment of invasive cervical cancer is affected by the stage of the disease, which is based on clinical evaluation. Microinvasive carcinoma of the cervix has limited metastatic potential and therefore is most likely curable by non-radical treatment. There is no standard management of stage Ib-IIa cervical carcinoma. Both radical surgery and radical radiotherapy have proven to be equally effective, but differ in associated morbidity and complications. Most often, stage Ib1 cervical cancer is treated by radical hysterectomy with pelvic lymphadenectomy. Laparoscopically assisted radical vaginal hysterectomy has shown similar efficacy and recurrence rates. Radical vaginal trachelectomy with laparoscopic pelvic lymphadenectomy may be an option in small cervical cancer where preservation of fertility is desired. There is lot of conflicting published work regarding the treatment of bulky stage Ib-IIa cervical cancer. While some centers are performing primary surgery as for Ib1 disease followed by tailored postoperative radiation with or without chemotherapy, the others are in favor of primary chemo-radiation therapy. Neoadjuvant chemotherapy followed by radical surgery has emerged as a possible alternative, which may improve a survival in patients with stage Ib2 disease. Concomitant chemoradiation is becoming a new standard in treatment of advanced disease, because it has been clearly shown to improve disease-free, progression-free and overall survival. Management of recurrent disease depends on previous treatment, site and extent of recurrence, disease-free interval and patient's performance status. CONCLUSIONS: Treatment decisions should be individualized and based on multiple factors including the stage of the disease, age, medical condition of the patient, tumor-related factors and treatment preferences, to yield the best cure with minimum complications. PMID- 16698224 TI - Gynaecological oncology perspective on management of the menopause. AB - Treatment for gynaecological cancer may ablate ovarian function through surgery (oophorectomy), radiotherapy or chemotherapy. Bilateral oophorectomy or surgical menopause results in an immediate menopause which may be intensely symptomatic in young women. Also oestrogen deficiency before the age of 40 increases the risk of osteoporosis and cardiovascular disease. This article will describe the use of oestrogen and non-oestrogen-based treatments as well as alternative and complementary therapies in gynaecological cancer survivors. AIMS: To review the use of oestrogen and non-oestrogen-based treatments as well as alternative and complementary therapies in gynaecological cancer survivors in whom ovarian function has been ablated through surgery (oophorectomy), radiotherapy or chemotherapy. METHODS: Literature search using PubMed for indexed articles. RESULTS: Ovarian, cervical, vaginal and vulval cancers are not oestrogen dependent conditions and oestrogen replacement is not contraindicated. However, there is some doubt with regard to endometrioid ovarian cancer and endometrial carcinoma is often listed in data sheets as an absolute contra-indication to hormone replacement therapy. Progestogens may be effective in treating vasomotor symptoms. A variety of agents are available for the prevention and treatment of osteoporosis and all except parathyroid hormone and strontium ranelate act mainly by inhibiting bone resorption. There is little scientific evidence that complementary and alternative therapies can help menopausal symptoms or conserve bone mass and there are no safety data in women with gynaecological cancer. CONCLUSIONS: Oestrogen-based therapies are the treatments of choice in young women with a premature menopause since oestrogen deficiency before the age of 40 increases the risk of osteoporosis and cardiovascular disease. PMID- 16698225 TI - [Assessment of patient satisfaction after regional anaesthesia in two institutions]. AB - OBJECTIVES: This study was designed to assess patient satisfaction after regional anaesthesia for limb surgery. METHODS: An anaesthesia satisfaction questionnaire was developed, validated (Kappa coefficient) and submitted to 314 patients operated in two institutions (one university hospital with anaesthesiology residents and one specialised in orthopaedics with experienced anaesthesiologists). Items explored were information modalities, pain and anxiety during procedure and global satisfaction rated with four levels (very satisfied [VS], satisfied [S], partially satisfied [PS], non-satisfied [NS]). Patients were interviewed by telephone at postoperative D1 and D8 by a pharmacist student not involved in the patient's care. RESULTS: Inspite of a high level of patient satisfaction at D8 (VS: 50%, S: 44%), some interesting aspects should be emphasised: a) sedation given before nerve block was not efficient to reduce anxiety and pain during procedure; b) VS levels decreased from D1 (56%) to D8 (50%) mainly because of late postoperative pain (after discharge) and discomforts; c) willingness to undergo the same nerve block again (294/314) was not correlated with patient's satisfaction since among PS and NS patients, a majority (9/15) wished for a block in case of renewed limb surgery; d) multivariate analysis showed that VS level was highly correlated with the quality of communication by the anaesthesiologist mainly for informations about pre and postoperative periods. No correlation was found with pain level during procedure; e) satisfaction levels were not different in the two institutions. CONCLUSION: This study has emphasised some important factors of patient satisfaction which were not sufficiently taken into account in our daily practice. PMID- 16698226 TI - [Rhabdomyolysis and paraphenylene-diamine poisoning]. AB - OBJECTIVE: The aim of this study is to describe clinical description, biological findings, outcome and prognostic factors of paraphenylene-diamine poisoning. PATIENTS AND METHODS: We report a cohort study spreaded over 6 years (1999-2004), realized in Medical Intensive Care Unit in Ibn-Rochd University Hospital at Casablanca (Morocco). This study included 315 patients admitted for paraphenylene diamine (PPD) poisoning. Diagnosis was based on: poisoning reported by the patient or his family, clinical data, biological findings and qualitative determination of PPD. Epidemiological parameters was obtained at admission. Every day, clinical and biological data, therapy and gravity scores were collected and a mean was calculated. RESULTS: 315 patients were admitted over this period. The mean age was 23+/-9 years. We noticed a clear female predominance (sex ratio=9.86). The intoxication was voluntarily aiming at autolysis in 93.3% of the cases. The patients were admitted at about 5+/-5.3 hours after the intoxication. The clinical chart was at first dominated by the respiratory and renal symptoms. The mean of CPK was 132,351.8+/-164,978 UI/l. The treatment was especially symptomatic. The mortality was 47%. The multivariate analysis concluded that acid urinary pH, hyperglycaemia, hard muscles, betamimetic drugs and MPM II>0.14 were associated with a poor prognosis. CONCLUSION: The PPD poisoning represents the first cause of toxic rhabdomyolysis in our context and responsible of high mortality. For that, it's necessary to control PPD trade, to inform the medical persons and a rapid management. PMID- 16698227 TI - [Maternal and foetal anaesthesia for ex utero intrapartum treatment (EXIT) procedure]. AB - The ex utero intrapartum treatment (EXIT) procedure is a surgical procedure maintaining utero-placental circulation during caesarean section. Anaesthetic implications are described: foetal transplacental anaesthesia to avoid first breathing and to permit surgical procedure on obstructed foetal airway, deep maternal haemodynamically stable anaesthesia to relax uterine smooth muscle during a long caesarean procedure but avoiding post-partum haemorrhage. Volatile anaesthesia with sevoflurane seems to be adequate for these aims. Two cases are described. PMID- 16698229 TI - [Microdialysis and brain injury: importance of animal study]. AB - Brain microdialysis allows the exploration of brain extracellular medium. This review discusses the main contribution of brain microdialysis for the knowledge of the pathophysiology of brain ischemia and trauma. We describe fundamental principle of microdialysis, limits, and validated metabolic parameters as the lactate/pyruvate ratio or glycerol. The interest to use microdialysis for testing metabolic hypothesis and potential scientific research ways will also be discussed. PMID- 16698230 TI - [Amniotic fluid embolism: successful evolution course of isolated disseminated intravascular coagulation and early biological diagnosis]. AB - Amniotic embolism is a sudden, unexpected and devastating complication of pregnancy. The diagnosis is usually made on the basis of clinical presentation after excluding differential diagnosis or at autopsy in the event of death of the parturient. We need to develop simple, non-invasive, sensitive tests for a reliable and early diagnosis. We report the case of a 34-year-old woman, who presented soon after delivery, an isolated disseminated intravascular coagulation with severe haemorrhage, an haemostatic hysterectomy was required. A 3370 g child was delivered by caesarean section. The patient survived without sequelae. The diagnosis of amniotic embolism was established by the presence of amniotic cells in the maternal central venous blood as well as in the bronchoalveolar fluid. PMID- 16698231 TI - [Validation of the french translated Richmond vigilance-agitation scale]. AB - OBJECTIVE: To test reliability of the Richmond Agitation Sedation Scale (RASS) after French translation. STUDY DESIGN: Prospective psychomotor evaluation study. PATIENTS AND METHODS: Two bilingual physicians performed the translation from English to French language. The French version was then translated to English by a bilingual non physician ("backtranslation") and compared to the original version. The translated scale was tested according to usual guidelines concerning the translation of neuropsychological evaluation tools. The French version of RASS was tested on the morning of every fifth or sixth day, and 30-min after any procedure, in 43 consecutive patients admitted to a 12-bed medical-surgical intensive care unit during two months, by a group of four observers: the principal investigator, two anaesthesiology residents and one pharmacology student. Inter-rater reliability was tested using the simple and weighted Kappa coefficients (with their 95% confidence interval). A consensual and detailed "operating instructions" guide of RASS, intended for medical and nursing staff, has been added in the Annexe section. RESULTS: Four hundred twenty measures were performed during 105 grouped observations. Reliability of the French version of RASS was substantial. Simple Kappa coefficient, testing the concordance between observers, was ranged from 0.72 (0.62-0.81) to 0.87 (0.79-0.94) and weighted Kappa coefficient from 0.95 (0.92-0.98) to 0.99 (0.98-0.99). CONCLUSIONS: The French translation of RASS was acceptable. The translated scale preserved substantial inter-rater reliability. Monitoring of vigilance status in intensive care settings can be performed with this reliable clinical tool. PMID- 16698232 TI - [Intracerebral haemorrhage, what's new?]. AB - Haemorrhagic stroke is frequent and associated with a high mortality and morbidity. Less than 30% of patients are still alive five years after onset and few patients regain functional independence. The worsening effect of anticoagulation has been demonstrated and the failure to rapidly normalize coagulation further increases haematoma expansion. In a recent phase II trial, recombinant activated factor VII given within 4 hours of stroke onset, reduced haematoma growth, mortality and disability. An aggressive blood pressure and intracranial pressure control early after the haemorrhage seems beneficial. A large prospective randomized study (the STICH trial) did not demonstrate any beneficial effect of surgery. PMID- 16698233 TI - Transcriptional network dynamics in macrophage activation. AB - Transcriptional regulatory networks govern cell differentiation and the cellular response to external stimuli. However, mammalian model systems have not yet been accessible for network analysis. Here, we present a genome-wide network analysis of the transcriptional regulation underlying the mouse macrophage response to bacterial lipopolysaccharide (LPS). Key to uncovering the network structure is our combination of time-series cap analysis of gene expression with in silico prediction of transcription factor binding sites. By integrating microarray and qPCR time-series expression data with a promoter analysis, we find dynamic subnetworks that describe how signaling pathways change dynamically during the progress of the macrophage LPS response, thus defining regulatory modules characteristic of the inflammatory response. In particular, our integrative analysis enabled us to suggest novel roles for the transcription factors ATF-3 and NRF-2 during the inflammatory response. We believe that our system approach presented here is applicable to understanding cellular differentiation in higher eukaryotes. PMID- 16698234 TI - The expression of the NR1-subunit of the NMDA receptor during mouse and early chicken development. AB - It has been suggested that homocysteine-induced defects are mediated by the inhibition of the N-methyl-d-aspartate (NMDA) receptor on neural crest cells. However, the majority of this work has been performed using the chicken embryo model. In an effort to better understand the molecular events involved a murine model of homocysteine-induced defects was sought. However, it has been previously shown that homocysteine failed to induce congenital defects in several strains of mouse. Therefore, in an effort to better understand the difference in the susceptibility between these two species we investigated the ontogeny of the NMDA receptor in the mouse and chicken. To determine the expression of the NMDA receptor we performed Western blot analysis using an antibody to the NR1-subunit of the NMDA receptor in both the chicken and mouse embryos. Further, we used RT PCR to determine the temporal expression of this subunit in the murine embryos from gestational day 8.5 to 18.5 to confirm our Western blot analysis. Results from these studies demonstrated that the expression of the NMDA receptor was expressed during the early stages of development in the chick embryo but neither the transcript nor the protein was detected in mouse embryos until later in development. These results demonstrate that during the stages of neurulation and/or early heart development the expression of the NR1-subunit of the NMDA receptor was not detected. The expression of this gene increased and was detectable by gestational days 14.5-15.5 and continued to increase in its expression until term. Therefore, these experiments suggest that homocysteine induced defects may be mediated via the NMDA receptor. PMID- 16698235 TI - Learning movement sequences with a delayed reward signal in a hierarchical model of motor function. AB - A key problem in reinforcement learning is how an animal is able to learn a sequence of movements when the reward signal only occurs at the end of the sequence. We describe how a hierarchical dynamical model of motor function is able to solve the problem of delayed reward in learning movement sequences using associative (Hebbian) learning. At the lowest level, the motor system encodes simple movements or primitives, while at higher levels the system encodes sequences of primitives. During training, the network is able to learn a high level motor program composed of a specific temporal sequence of motor primitives. The network is able to achieve this despite the fact that the reward signal, which indicates whether or not the desired motor program has been performed correctly, is received only at the end of each trial during learning. Use of a continuous attractor network in the architecture enables the network to generate the motor outputs required to produce the continuous movements necessary to implement the motor sequence. PMID- 16698236 TI - Mineralization and mechanical properties of the canine mandible distraction wound following acute molding. AB - To investigate the influence of an acute single step callus manipulation immediately after distraction on mechanical properties and mineralization of the regenerate, custom made distraction devices were bilaterally placed in the mandibular angle of 15 beagle dogs, allowing to simultaneously compress and stretch the regenerate after completed linear distraction. The animals were divided in three groups (n=5): Group 1 and 2 underwent manipulation of the regenerate, group 3 remained in a linear position. After 42 (group1) and 90 (group 2 and 3) days of consolidation the animals were sacrificed. The mechanical properties were assessed in an Instron testframe and bone density quantified by quantitative computed tomography and three- dimensionally assessed (Scion Image processing and analysis software). After 6 weeks of consolidation 25% of the specimens reached a stiffness which was >/=90% of the mean values of the unoperated reference hemi-mandibles. After a 13 week consolidation period, 62.5% were as stiff as the referenced specimens. Manipulated regenerates, allowed to heal under stable conditions for 13 weeks, had the same mechanical properties as specimens that underwent pure linear distraction. A temporary but not significant delay of osseous healing had to be postulated for the stretched zone after 6 weeks, indicating this area to be more critical than the compressed area. PMID- 16698237 TI - A simple formula for individualising ceftazidime dosage administered by continuous infusion in patients with haematological malignancies. AB - A formula is proposed for individualising ceftazidime dosage administered by continuous infusion in patients with haematological malignancies. Sixty patients were retrospectively randomised into Group A (n=30) to establish the formula and Group B (n=30) to evaluate this formula. Individual ceftazidime clearances were estimated from the ratio between the rate of infusion and plasma concentration at steady state. In Group A, ceftazidime clearance was significantly correlated with creatinine clearance. From this result, a formula (rate of infusion (g/day)=0.00133x[creatinine clearance (mL/min)]x[target concentration at steady state (mg/L)]) is proposed. This formula provided consistent estimations of ceftazidime plasma concentrations in Group B and should help clinicians to define the optimum ceftazidime dosage, particularly in patients with disturbed renal function. PMID- 16698238 TI - Evaluation of dalbavancin in combination with nine antimicrobial agents to detect enhanced or antagonistic interactions. AB - Dalbavancin is a potent, once-weekly administered lipoglycopeptide that is active against a broad spectrum of Gram-positive species. Synergy studies were performed with dalbavancin and each of nine antimicrobial agents (90 tests in total) representing nine antimicrobial classes using the broth microdilution checkerboard method to establish in vitro interactive categories. Antagonism was not observed between dalbavancin and any of the antimicrobials tested. However, dalbavancin was synergistic or partially synergistic with oxacillin for staphylococci, including methicillin-resistant strains, vancomycin-intermediate Staphylococcus aureus and enterococci, a significant finding that warrants further investigation to establish its potential clinical relevance. PMID- 16698239 TI - Temperature-pH sensitivity of bovine serum albumin protein-microgels based on cross-linked poly(N-isopropylacrylamide-co-acrylic acid). AB - Monodispersed poly(N-isopropylacrylamide-co-acrylic acid) (PNIPAM/AAc) microgels with various contents of acrylic acid (AAc) and cross-linker N,N' methylenebisacrylamide (MBAAm) were synthesized by surfactant-free emulsion polymerization. The microgels exhibited affinity of binding bovine serum albumin (BSA) with increase of AAc contents. The adsorption amount of BSA was strongly dependent on suspension pH. The maximum adsorption was observed at around pH 4.0, while both low pH and basic conditions induced the rather low adsorption. In addition, AFM images showed that the microgel particles underwent bridging aggregation by loading BSA. The resultant BSA-microgel particles became more hydrophobic after loading BSA. Importantly, the adsorption of BSA onto PNIPAM/AAc microgel was found to alter the swelling/deswelling behavior of the resultant microgel suspensions. With the increase of BSA concentration, the volume phase transition temperature of BSA-microgel particles shifted to the lower temperature. PMID- 16698240 TI - [Cardiac output in the child: should it be investigated? And how?]. PMID- 16698241 TI - [Migrain in children]. PMID- 16698242 TI - [Ethical approach to clinical uncertainty]. PMID- 16698243 TI - [Osteogenesis imperfecta or child abuse]. PMID- 16698244 TI - [Calcium, vitamin D, physical activity and bone mineral content of sport children and adolescents]. PMID- 16698245 TI - [Imaging of back pain in children]. PMID- 16698246 TI - [Magnetic resonance urography in children]. PMID- 16698247 TI - [Physical fitness in children with asthma]. AB - The exercise limitation in a child with asthma is the result of several factors: the lack of ventilatory reserve, muscle deconditioning, and cardiac limitation. The severity of the airway obstruction is a major determinant of physical fitness. Aerobic and anaerobic fitness are equally affected. Both types of fitness can be improved by specific training programs. PMID- 16698248 TI - [French regulation for radiation dose control in paediatric radiography]. PMID- 16698249 TI - [Surgical pitfalls in adolescents]. PMID- 16698250 TI - [Pathophysiology and genetics of deafness]. PMID- 16698251 TI - [Labelling, claims and infant or young child special foods]. PMID- 16698252 TI - [Localization of nerves in pediatric regional anesthesia]. PMID- 16698253 TI - [Family planning: visit without parents]. PMID- 16698254 TI - [Asthma in children: how to improve adherence]. PMID- 16698255 TI - [Fetal neurology: conditions of diagnostic uncertainty]. PMID- 16698256 TI - Characterization and biological activity of the main flavonoids from Swiss Chard (Beta vulgaris subspecies cycla). AB - The molecular components of a phenolic fraction (P2), obtained from liquid chromatography of a Swiss Chard (Beta vulgaris subsp. cycla) extract, were identified using HPLC-ESI-MS/MS. The primary P2 components were: vitexin-2''O rhamnoside, its demethylated form 2''-xylosylvitexin, isorhamnetin 3 gentiobioside, and rutin. P2 "in toto" and the single components were characterized for antioxidant capacity, antimitotic activity on MCF-7 human breast cancer cells and for toxicity to human lymphocytes and macrophages. P2 inhibited MCF-7 cell proliferation (IC(50) value = 9 microg/ml) without inducing apoptosis, showed no toxicity to human lymphocytes and slight toxicity to macrophages. Vitexin-2''O-rhamnoside strongly inhibited DNA synthesis in MCF-7 cells, whereas 2''-xylosylvitexin and isorhamnetin 3-gentiobioside were activators; combinations of activators and inhibitors maintained the over-all inhibitory effect. PMID- 16698257 TI - The effects of desferrioxamine and quercetin on hepatic ischemia-reperfusion induced renal disturbance. AB - BACKGROUND: The aim of this study was to analyze the effects of 45min of hepatic ischemia and 1h of reperfusion on renal oxidative stress parameters, on renal tissue damage, and the role of Desferrioxamin (Dfx) and Q on these parameters. METHODS: Thirty Wistar albino rats were randomized to five groups. Group I was the control group. Group II received no treatment. Groups III and IV received intramuscular injections of desferrioxamine (100mg/kg) and quercetin (50mg/kg), respectively. Group V was administered Dfx and quercetin in combination. After treatment for 3 days, groups II, III, IV, and V were exposed to total hepatic ischemia for 45min. Plasma alanine aminotransferase levels, renal malondialdehyde and reduced glutathione (GSH) activities were measured after reperfusion for 1h. Histopathological and ultrastructural analysis of renal tissues was carried out. RESULTS: Plasma creatinine and BUN levels were markedly increased in the IR group and pretreated groups. Kidney MDA increased in the IR group, Q and Dfx+Q significantly decreased kidney MDA Kidney GSH levels markedly decreased in the IR group, Dfx significantly increased kidney GSH. No evidence of overt injury was observed in any renal tissue under light and electron microscopy. CONCLUSIONS: Our data demonstrated that 45min of hepatic ischemia and 1h of reperfusion may alter renal functions and may cause oxidative stress on renal tissue. Q and Dfx seem to have a beneficial effect via the GSH system and modulation of MDA levels. PMID- 16698258 TI - Observational study to compare the clinical efficacy of the natural surfactants Alveofact and Curosurf in the treatment of respiratory distress syndrome in premature infants. AB - INTRODUCTION: Natural surfactants have been shown to be superior to synthetic surfactants in the treatment of neonatal respiratory distress syndrome (RDS). In Germany, Alveofact (A) and Curosurf (C) are the most frequently used natural surfactant preparations. The aim of this retrospective, observational study was to compare the effects of A and C on gas exchange and outcome in premature infants. METHODS: During a 5-year period in our neonatal intensive care unit (NICU), 187 premature infants were treated with surfactant, with 82 receiving A and 105 receiving C. We recorded F(I)O(2) and gas exchange (PaO(2)/F(I)O(2) ratio, PaCO(2), SaO(2)) during the first 72h after surfactant application and the incidence of outcome parameters at day 28 (bronchopulmonary dysplasia (BPD), intraventricular hemorrhage (IVH grade III or IV), patent ductus arteriosus (PDA), pneumothorax, necrotizing enterocolites (NEC) and death). The differences between the patient groups were assessed by ANOVA or the calculation of relative risks. Bonferroni correction was used for multiple comparisons. RESULTS: There were no statistically significant differences between infants treated with A and C in mean gestational age (28.4 vs. 28.4 weeks), birth weight (1210 vs.1258 g) and time of first surfactant application (60 vs. 90 min postnatal). We observed no significant between group differences in course of F(I)O(2) and blood gases, or in incidence at day 28 of BPD (41.7% vs. 42.8%), IVH III/IV (18.3% vs. 14.3%), pneumothorax (9.8% vs. 4.8%), PDA (23.2% vs. 21.9%), PVL (7.3% vs. 9.5%) and death (17% vs. 17.1%). There were also no statistically significant differences in the subgroup of infants <28 weeks. The lower incidence of NEC in A compared with C (1.2% vs. 10.5%, P=0.01) was not statistically significant after Bonferroni correction. CONCLUSION: Independent of gestational age no significant difference in the clinical efficacy of A and C was observed. PMID- 16698259 TI - Tiotropium in COPD patients not previously receiving maintenance respiratory medications. AB - INTRODUCTION: Use of maintenance bronchodilator therapy is currently recommended in symptomatic patients with Chronic obstructive pulmonary disease (COPD) and in those with Stage II or greater COPD as defined by the Global Initiative for Chronic Obstructive Lung Disease (GOLD). Because no prospective data describe when rescue therapy alone is insufficient or the optimal time to start maintenance therapy, it is unclear whether maintenance therapy has benefits in milder disease. To explore potential benefits we asked: Does once-daily tiotropium improve lung function, health status, and/or symptoms in "undertreated" COPD patients (i.e., those who are not receiving maintenance bronchodilator therapy) or patients considered by their health care providers as having milder disease? METHODS: A post-hoc analysis of data from COPD patients participating in two, 1-year, placebo-controlled trials with tiotropium was performed. Patients were defined as "undertreated" if they received no respiratory medication or only as-needed short-acting beta-agonists prior to enrollment. Measures included serial spirometry, Transition Dyspnea Index (TDI), and St. George's Respiratory Questionnaire (SGRQ). RESULTS: Of 921 patients enrolled, 218 (23.7%) were "undertreated": 130 received tiotropium; 88 received placebo. Demographics for the two treatment groups were comparable. Tiotropium treated patients had significantly improved forced expiratory volume in 1s (FEV1) and forced vital capacity (FVC) compared with patients using placebo on all study days. Additionally, TDI and SGRQ scores significantly improved with tiotropium compared with placebo. CONCLUSIONS: Once-daily tiotropium provides significant improvement in lung function, health status, and dyspnea when used as maintenance therapy in undertreated COPD patients who were not previously receiving maintenance bronchodilator therapy. PMID- 16698260 TI - Enhanced surface plasmon resonance with the modified catalytic growth of Au nanoparticles. AB - The catalytic growth of Au nanoparticles (AuNPs) has been employed in several analytical methods for improving the detection sensitivity, or integrated with the enzyme reactions for the quantitative detection of the respective substrates. However, the catalytic growth of Au nanoparticles do not work in some situations, such as surface plasmon resonance (SPR), electrochemistry, where metal matrices were used, because metal matrices used in these techniques, e.g. Au, are susceptible to metal deposition, which increased the background seriously. In this work, a SiO(2) layer was vapor-deposited on the gold film. The inhibition of metal deposition by this SiO(2) layer was investigated by SPR sensor. The results showed that the SiO(2) layer could avoid the deposition of metal on Au film. With the low background achieved by SiO(2)-coated Au films, sensitive detection of DNA hybridization using the catalytic growth of Au nanoparticles enhanced SPR was demonstrated. The work described here maybe helpful for the development of sensitive bioanalytical methods. PMID- 16698261 TI - MHC homologs in the nervous system--they haven't lost their groove. AB - Major histocompatibility complex (MHC) molecules have been implicated in a number of non-immune roles in the central nervous system, particularly in synaptic development and plasticity. The discovery of M10 (50% sequence identity to classical MHC molecules) proteins expressed in the vomeronasal organ adds to the list of non-traditional roles of MHC homologs. M10 molecules associate with the V2R class of vomeronasal receptors, a family of G-protein coupled receptors thought to function as pheromone receptors. Recent studies showing that classical MHC-binding peptides activate V2R-expressing neurons offer tempting clues that M10s might participate directly in the recognition of pheromone ligands, but M10 proteins do not bind to these peptides with significant affinity. Instead of presenting MHC-binding peptides, M10s might function as molecular chaperones to V2R receptors or more generally as modulators of neuronal function, as demonstrated elsewhere in the brain for classical MHC molecules. PMID- 16698262 TI - Calcium-permeable AMPA channels in neurodegenerative disease and ischemia. AB - Compelling evidence supports contributions of glutamate receptor overactivation ('excitotoxicity') to neurodegeneration in both acute conditions, such as stroke, and chronic neurodegenerative conditions, such as amyotrophic lateral sclerosis. However, anti-excitotoxic therapeutic trials, which have generally targeted highly Ca2+ permeable NMDA-type glutamate channels, have to date failed to demonstrate impressive efficacy. Whereas most AMPA type glutamate channels are Ca2+ impermeable, an evolving body of evidence supports the contention that relatively unusual Ca2+ permeable AMPA channels might be crucial contributors to injury in these conditions. These channels are preferentially expressed in discrete neuronal subpopulations, and their numbers appear to be upregulated in amyotrophic lateral sclerosis and stroke. In addition, unlike NMDA channels, Ca2+ permeable AMPA channels are not blocked by Mg2+, but are highly permeable to another potentially harmful endogenous cation, Zn2+. The targeting of these channels might provide efficacious new avenues in the therapy of certain neurological diseases. PMID- 16698263 TI - Anaerobic digestion of cattail with rumen culture in the presence of heavy metals. AB - The anaerobic digestion of cattail by rumen cultures in the presence of Cu(II), Cd(II) or Cr(VI) was investigated in this study. Three cases were respectively observed for the different metal dosages: promoted cattail degradation and methane production at a low heavy metal concentration, e.g., Cu(II) 2.4 mg/l, Cd(II) 1.6 mg/l, Cr(VI) 4.0 mg/l; reduced cattail degradation efficiency and methane production at a middle metal level; a severe inhibition to the cattail degradation at a high heavy metal dosage. The inhibition kinetics of Cu(II) on the digestion of cattail by rumen cultures was also analyzed and a simplified Andrews equation was used to describe such an inhibition. The inhibition constants for Cu(II) on the degradation of cattail, production of volatile fatty acids and formation of methane were estimated as 7.4, 9.5 and 6.4 mg/l, respectively. Comparative experimental results suggest that the order of toxicity degree of the tested metals on the rumen cultures was: Cd(II) > Cu(II) > Cr(VI). PMID- 16698264 TI - Discovery of 3,5-bis(trifluoromethyl)benzyl L-arylglycinamide based potent CCR2 antagonists. AB - Systematic modification of a screening lead yielded a class of potent glycinamide based CCR2 antagonists. The best compound (55, (2S)-N-[3,5 bis(trifluoromethyl)benzyl]-2-{[2-(1-piperidinyl)ethyl]amino}-2-(3 thienyl)acetamide) displayed good binding affinity (IC50=30 and 39 nM) toward human monocytes and CHO cell expressing human CCR2b, respectively. Functionally, it blocked MCP-1 (CCL2)-induced calcium mobilization (IC50=50 nM) and chemotaxis mediated through the CCR2 receptor (9.6 nM). It is selective against other chemokine receptors tested. PMID- 16698265 TI - Azadideoxyadenosine: synthesis, enzymology, and anti-HIV activity. AB - Synthesis of an azanucleoside, a new analogue of dideoxyadenosine, is described. This compound is only slowly deaminated by mammalian adenosine deaminase and it is a substrate for adenosine kinase. It exhibits in vitro anti-HIV activity. PMID- 16698266 TI - Putative therapeutic agents for the learning and memory deficits of people with Down syndrome. AB - Mental retardation is the most common and debilitating condition for individuals with Down syndrome (DS). The hyper-activation of DYRK1A by overexpression causes significant learning and memory deficits in DS-model mice. Thus far, no mechanism based drug has been developed to address this. After a combination of in silico and in vitro screenings, two DYRK1A inhibitors were isolated that are active in a cell-based assay. Further optimization could lead to a novel drug discovery that could address DS learning and memory deficits. PMID- 16698267 TI - Unraveling the mechanisms of synaptotagmin and SNARE function in neurotransmitter release. AB - SNARE proteins and synaptotagmin are key components of the complex machinery that controls Ca(2+)-triggered neurotransmitter release but their mechanisms of action are under debate. Recent research has shed light on which biochemical and/or biophysical properties underlie SNARE and synaptotagmin function. SNARE proteins most likely have a role in membrane fusion owing to their ability to bring the synaptic vesicle and plasma membranes together and to perturb lipid bilayers through their transmembrane regions. Synaptotagmin acts as a Ca(2+) sensor and might cooperate with the SNAREs in accelerating fusion by binding simultaneously to the two membranes. However, recent research has strongly challenged the validity of models proposing that the SNAREs (with or without synaptotagmin) constitute "minimal membrane fusion machineries" and has emphasized the essential nature of other proteins for exocytosis. Understanding the functions of these proteins will be crucial to reach a faithful description of the mechanisms of membrane fusion and neurotransmitter release. PMID- 16698268 TI - Whither critical care outreach? PMID- 16698269 TI - Ligands with a 3,3-diphenylpentane skeleton for nuclear vitamin D and androgen receptors: Dual activities and metabolic activation. AB - Ligands possessing dual vitamin D(3) (VD(3))-agonistic and androgen-antagonistic activities with various activity spectra were prepared based on a substituted 3,3 diphenylpentane (DPP) skeleton. Among the compounds, (R,S)-DPP-1023 [(R,S)-7b] and (S,S)-DPP-0123 [(S,S)-7c] showed the most potent vitamin D(3)-agonistic activity [with potency comparable to that of 1alpha,25-dihydroxyvitamin D(3) (1,25-VD(3))] and nuclear androgen receptor (AR)-binding activity (with higher affinity than that of hydroxyflutamide), respectively. Metabolic activation (reduction of the carbonyl group) of pivaloyl analogs [DPP-1113 (3a), DPP-1013 (3b), DPP-0113 (3c), and DPP-0013 (3d)] in HL-60 cells was found to be necessary for binding to nuclear vitamin D(3) receptor (VDR). PMID- 16698270 TI - Synthesis and antidepressant-like action of stereoisomers of imidobenzenesulfonylaziridines in mice evaluated in the forced swimming test. AB - The present study describes the chemical synthesis and pharmacological evaluation of a new series of eleven compounds stereoisomers of imidobenzenesulfonylaziridines in the forced-swimming test (FST) in mice. The pharmacological results of these compounds show that six of them, given intraperitoneally, reduced the immobility time of mice evaluated in the FST, an antidepressant-like profile of action similar to imipramine, a well-known standard antidepressant drug used for comparison, without compromising the animals' motor performance. The putative antidepressant-like action demonstrated here indicates their viability for the development of new therapeutic options for the treatment of depression. PMID- 16698271 TI - Novel benzofuran inhibitors of human mitogen-activated protein kinase phosphatase 1. AB - Protein tyrosine phosphatases have a central role in the maintenance of normal cellular functionality. For example, PTP1B has been implicated in insulin resistance, obesity, and neoplasia. Mitogen-activated protein kinase phosphatase 1 (MKP-1 or DUSP1) dephosphorylates and inactivates mitogen-activated protein kinase (MAPK) substrates, such as p38, JNK, and Erk, and has been implicated in neoplasia. The lack of readily available selective small molecule inhibitors of MKP family members has severely limited interrogation of their biological role. Inspired by a previously identified inhibitor (NSC 357756) of MKP-3, we synthesized seven NSC 357756 congeners, which were evaluated for in vitro inhibition against several protein phosphatases. Remarkably, none displayed potent inhibition against MKP-3, including the desamino NSC 357756 analog NU-154. Interestingly, NU-154 inhibited human PTP1B in vitro with an IC(50) value of 24 +/- 1 microM and showed little inhibition against Cdc25B, MKP-1, and VHR phosphatases. NU-126 [2-((E)-2-(5-cyanobenzofuran-2-yl)vinyl)-1H-indole-6 carbonitrile] inhibited MKP-1 and VHR in vitro but was less active against human MKP-3, Cdc25B, and PTP1B. The inhibition of MKP-1 by NU-126 was independent of redox processes. The benzofuran substructure represents a new potential scaffold for further analog development and provides encouragement that more selective and potent inhibitors of MKP family members may be achievable. PMID- 16698272 TI - Carborne gamma-ray spectrometry. Calibration and applications. AB - Calibration of carborne gamma-ray spectrometry systems for (137)Cs is carried out with a source successively placed at 791 positions within an area of 34 m x 62 m. A computer model supplements the measurements. Hereby a sensitivity map for a surface contamination is generated as well as line and area sensitivities. Another model converts surface sensitivity to sensitivity for a deep contamination. Use of the sensitivity map for a non-homogeneous distribution of (137)Cs is demonstrated. Applications of line sensitivities for special tasks are discussed. PMID- 16698273 TI - Excitation functions for the production of 82Sr by proton bombardment of natRb at energies up to 100 MeV. AB - The excitation functions for the production of (82)Sr and other radionuclides produced in the proton bombardment of rubidium were measured by means of the activation technique. Stacks were assembled from RbCl targets, aluminium and copper monitor foils and bombarded with protons of energy up to 100 MeV and nominal current 0.1 microA. The measured data were compared with the theoretical calculations obtained by means of ALICE-IPPE, and also with previously published data. The measured data sets exhibit good agreement at incident energies below 45 MeV and greater than 60 MeV but show large discrepancies in the energy range between these values. The ALICE-IPPE calculations evidently overestimate all reported experimental cross sections, in particular overestimating the present data by a factor of 1.6. Notwithstanding the fact that there are discrepancies in the cross sections, good agreement is found with regards to the shape of the excitation function. PMID- 16698274 TI - Hormonal and cytokine regulation of early implantation. AB - Implantation of the blastocyst into the endometrium is a delicately controlled process and a prerequisite for the furtherance of the mammalian species. A complex network of molecules is involved in preparing both the endometrium and blastocyst for a successful interaction. However, the exact molecular steps are poorly understood. Studies so far have shown that disruption of certain pathways results in fertility defects. Impaired implantation is currently considered to be the most important limiting factor for the establishment of viable pregnancies in assisted reproduction. It is expected that elucidating the molecular background of the process will enable accurate diagnosis and effective treatment of infertility. PMID- 16698275 TI - Combined administration of G-CSF and GM-CSF stimulates monocyte-derived pro angiogenic cells in patients with acute myocardial infarction. AB - Mobilization of endothelial progenitor cells has been suggested to contribute to neo-vascularization of ischemic organs. Aim of this study was to investigate whether the combination of granulocyte colony stimulating factor (G-CSF) and granulocyte-macrophage (GM)-CSF may influence the expansion of circulating KDR+ cells in patients with acute myocardial infarction (AMI). KDR+ cells significantly increased in peripheral blood of AMI patients treated with G-CSF and GM-CSF compared to untreated patients. This KDR+ cells population was CD14+ but not CD34+ or CD133+. CD14+/KDR+ cells were also obtained in vitro by culturing mononuclear cells from healthy donors in a Rotary Cell Culture System in the presence of G-CSF + GM-CSF, but not of the individual growth factors. CD14+/KDR+ cells, obtained from patients or from in vitro culture, co-expressed hematopoietic (CD45, CD14) and endothelial markers (CD31, CD105, and VE cadherin). CD14+/KDR+, but not CD14+/KDR- cells, stimulated the organization of human microvascular endothelial cells into capillary-like structures on Matrigel both in vitro and in vivo. The combination of G-CSF and GM-CSF induced a CD14+/KDR+ cell population with potential pro-angiogenic properties. PMID- 16698276 TI - Interleukin-10 or tumor necrosis factor-alpha polymorphisms and the natural course of hepatitis C virus infection in a hyperendemic area of Japan. AB - We investigated the effects of polymorphisms in interleukin (IL)-10 and tumor necrosis factor (TNF)-alpha on the natural course of hepatitis C virus (HCV) infection in a community-based population in Japan. A total of 460 anti-HCV antibody seropositive individuals were classified into two groups, those who were positive or negative for HCV RNA. In HCV RNA-positive individuals with at least four annual alanine aminotransferase (ALT) measurements taken between 1993 and 2003, 74 exhibited persistently normal ALT levels, while 211 had one or more elevated ALT level tests. We examined the relationships between polymorphisms in the genes encoding IL-10 (-1082, -819, -592) or TNF-alpha (-308, -238) and HCV clearance, ALT abnormalities, or serum level of type IV collagen 7S, a marker of hepatic fibrosis. These polymorphisms were equally distributed among the patient subgroups with differential HCV RNA clearances or ALT abnormalities. Serum levels of type IV collagen 7S, however, were significantly higher in individuals with an A at position -238 or -308 in the TNF-alpha gene promoter than in individuals lacking these polymorphisms. We conclude that, while the relationships between inherited variations in IL-10 or TNF-alpha expression are not associated with alterations in HCV clearance or ALT levels, TNF-alpha polymorphisms may be associated with hepatic fibrosis. PMID- 16698277 TI - Nafion-coated bismuth film and nafion-coated mercury film electrodes for anodic stripping voltammetry combined on-line with ICP-mass spectrometry. AB - Nafion-coated bismuth film electrodes (NCBFEs) and Nafion-coated mercury film electrodes (NCMFEs) were used to electrochemically preconcentrate metal analytes for subsequent analysis by inductively coupled plasma-mass spectrometry (ICP-MS). Either type of electrodes is part of a thin-layer electrochemical flow cell that is positioned upstream of a microconcentric nebulizer for the ICP-MS. Performances of these electrodes were compared in terms of the analytical "figures of merit" (e.g., dynamic ranges, reproducibility, hydrodynamic stability, and elimination of matrix effects detrimental to ICP-MS). The coupled technique (ASV-ICP-MS) is found to possess a wide dynamic range (at least 4 to 5 orders of magnitude) and to be reproducible. Both electrodes are much more stable than the thin mercury film electrode (TMFE) traditionally used for ASV-ICP-MS, with the lifetime of the NCBFE exceeding 8 h. Adopting these electrodes for ASV ICP-MS overcomes the problems associated with a TMFE, the erosion of which decreases the sample throughput, affects the analysis precision, and contaminates conventional glass nebulizers and spray chambers of the spectrometer. The medium exchange procedure inherent in ASV is successfully implemented with a two-valve flow injection system for the accumulation of trace Cd2+ into the electrode from a certified seawater sample, followed by stripping Cd into a solution that is compatible to the ICP-MS operation. PMID- 16698278 TI - Ion trap collision-induced dissociation of human hemoglobin alpha-chain cations. AB - Multiply protonated human hemoglobin alpha-chain species, ranging from [M + 4H]4+ to [M + 20H]20+, have been subjected to ion trap collisional activation. Cleavages at 88 of the 140 peptide bonds were indicated, summed over all charge states, although most product ion signals were concentrated in a significantly smaller number of channels. Consistent with previous whole protein ion dissociation studies conducted under similar conditions, the structural information inherent to a given precursor ion was highly sensitive to charge state. A strongly dominant cleavage at D75/M76, also noted previously in beam type collisional activation studies, was observed for the [M + 8H]8+ to [M + 11H]11+ precursor ions. At lower charge states, C-terminal aspartic acid cleavages were also prominent but the most abundant products did not arise from the D75/M76 channel. The [M + 12H]12+-[M + 16H]16+ precursor ions generally yielded the greatest variety of amide bond cleavages. With the exception of the [M + 4H]4+ ion, all charge states showed cleavage at the L113/P114 bond. This cleavage proved to be the most prominent dissociation for charge states [M + 14H]14+ and higher. The diversity of dissociation channels observed within the charge state range studied potentially provides the opportunity to localize residues associated with variants via a top-down tandem mass spectrometry approach. PMID- 16698279 TI - Electrochemically-induced reactions of hexafluorophosphate anions with water in negative ion electrospray mass spectrometry of undiluted ionic liquids. AB - The influence of water on the observed gas-phase population of negative ions in electrospray mass spectrometry was studied for the undiluted ionic liquid 1,3 butyl-methyl-imidazolium hexafluorophosphate (BMIM+PF6-). During the electrospray process, electrolytic reduction of water enhances the production of tetrafluorophosphate (F4PO-), which undergoes further reactions to produce difluorophosphate (F2PO2-) anions. These anions are observed in addition to the pre-existing hexafluorophosphate anion. The apparent substitution of two fluorine atoms with one oxygen is attributed to a series of reactions initiated by hydrolysis of hexafluorophosphate. This hydrolysis reaction was enhanced by the addition of hydroxide, formed via the hydrolysis of water or through the addition of ammonium hydroxide. The formation of FxPOy- was studied as a function of the electrospray current and solution flow rate. The mass spectral response shows a quantitative logarithmic relationship between SigmaFxPOy- signal intensities (adjusted for mole equivalents of H2O required) and the amount of water present, against which the water content could be rapidly assessed. Results were found to be comparable to Karl Fischer titration data. PMID- 16698280 TI - Focus in honor of Gary J. Van Berkel, 2005 Biemann Medal awardee. PMID- 16698281 TI - Epigenetic silencing of 14-3-3sigma in cancer. AB - The 14-3-3sigma gene is a direct target of the p53 tumor suppressor and its product inhibits cell cycle progression. Recently, a proteomic analysis revealed that 14-3-3sigma regulates additional cellular processes relevant to carcinogenesis, as migration and MAP-kinase signalling. The expression of 14-3 3sigma is down-regulated by CpG methylation in several types of human cancer, among them prostate, lung, breast and several types of skin cancer. The epigenetic inactivation of 14-3-3sigma occurs at an early stage of tumor development and may allow evasion from senescence and promote genomic instability. In the future the detection of CpG methylation of 14-3-3sigma may be used for diagnostic and prognostic purposes. PMID- 16698282 TI - Crystal structure of the actin-binding domain of alpha-actinin 1: evaluating two competing actin-binding models. AB - Alpha-actinin belongs to the spectrin family of actin crosslinking and bundling proteins that function as key regulators of cell motility, morphology and adhesion. The actin-binding domain (ABD) of these proteins consists of two consecutive calponin homology (CH) domains. Electron microscopy studies on ABDs appear to support two competing actin-binding models, extended and compact, whereas the crystal structures typically display a compact conformation. We have determined the 1.7A resolution structure of the ABD of alpha-actinin 1, a ubiquitously expressed isoform. The structure displays the classical compact conformation. We evaluated the two binding models by surface conservation analysis. The results show a conserved surface that spans both domains and corresponds to two previously identified actin-binding sites (ABS2 and ABS3). A third, and probably less important site, ABS1, is mostly buried in the compact conformation. However, a thorough examination of existing structures suggests a weak and semi-polar binding interface between the two CHs, leaving open the possibility of domain reorientation or opening. Our results are consistent with a two-step binding mechanism in which the ABD interacts first in the compact form observed in the structures, and then transitions toward a higher affinity state, possibly through minor rearrangement of the domains. PMID- 16698283 TI - Influence of selected Indian immunostimulant herbs against white spot syndrome virus (WSSV) infection in black tiger shrimp, Penaeus monodon with reference to haematological, biochemical and immunological changes. AB - Immunostimulants are the substances, which enhance the non-specific defence mechanism and provide resistance against the invading pathogenic micro-organism. In order to increase the immunity of shrimps against the WSSV, the methanolic extracts of five different herbal medicinal plants like Cyanodon dactylon, Aegle marmelos, Tinospora cordifolia, Picrorhiza kurooa and Eclipta alba were selected and mixed thoroughly in equal proportion. The mixed extract was supplemented with various concentrations viz. 100 (A), 200 (B), 400 (C), and 800 (D) mgkg(-1) through artificial diets individually. The prepared diets (A-D) were fed individually to WSSV free healthy shrimp Penaeus monodon with an average weight of 8.0+/-0.5g for 25 days. Control diet (E), devoid of herbal extract was also fed to shrimps simultaneously. After 25 days of feeding experiment, the shrimps were challenged with WSSV, which were isolated and propagated from the infected crustaceans. The shrimps succumbed to death within 7 days when fed on no herbal immunostimulant diet (E). Among the different concentrations of herbal immunostimulant supplemented diets, the shrimps fed on diet D (800mgkg(-1)) significantly (P<0.0001) had more survival (74%) and reduction in the viral load. Also the better performance of haematological, biochemical and immunological parameters was found in the immunostimulant incorporated diets fed shrimps. The present work revealed that the application of herbal immunostimulants will be effective against shrimp viral pathogenesis and they can be recommended for shrimp culture. PMID- 16698284 TI - Persistence of betanodavirus in Barramundi brain (BB) cell line involves the induction of Interferon response. AB - The BB cell line derived from the brain tissue of a barramundi (Lates calcarifer) that survived nervous necrosis virus (NNV) infection is persistently infected with NNV. To elucidate whether interferon (IFN) plays a role in the mechanism of NNV-persistent infection in BB cell line, a virus-negative control cell line was obtained by treating BB cells with NNV-specific rabbit antiserum for 5 subcultures. After the treatment, NNV titer or RNA or capsid protein was no longer detected in the cured BB (cBB) cells. Expression of Mx gene, encoding a type I IFN-inducible antiviral protein, was found in BB cells and cBB cells following NNV infection, but not in NNV-free cBB cells. Moreover, expression of Mx gene and antiviral activity against NNV were induced in cBB cells by the treatment with MAb-neutralized BB cell supernatant. Furthermore, NNV persistent infection was induced again in cBB cell culture if multiplicity of infection (MOI) was low (< or = 1). These experimental results indicated that IFN-like cytokines existed in the culture supernatant of BB cells, and IFN-induced response played an important role in protecting the majority of cells from virus lytic infection and regulating NNV persistence in the BB cell line. PMID- 16698285 TI - Moderate alterations in lower limbs muscle temperature do not affect postural stability during quiet standing in both young and older women. AB - Older adults demonstrate increased amounts of postural sway, which may ultimately lead to falls. Temperature is known to have a profound effect on the performance of the neuromuscular system which could have important implications on motor control. It is, therefore, of interest to investigate if the age-related decline in postural stability could be affected by changes in local limbs temperature. The present study investigated the effects of localized warming and cooling on postural sway in nine young (22+/-3 years) and nine older (73+/-3 years) women. Postural sway was assessed, using a single force platform, during quiet standing at three muscle temperature conditions: control (34.2+/-0.2 degrees C), cold (31.3+/-0.3 degrees C) and warm (37.0+/-0.1 degrees C). Two stances were evaluated, the Romberg (large support base) and modified Tandem (narrow support base), under both eyes-open and eyes-closed conditions. Root mean square (RMS), mean velocity (MV), sway area (SA) and mean power frequency (MPF) were calculated from the centre of pressure (COP) displacement. Neither warming nor cooling significantly affected any of the postural parameters which were, however, all higher (P<0.05) in the older group than the young group in all conditions. This study demonstrated that, in quiet standing conditions, a moderate variation (+/-3 degrees C) in lower limbs temperature does not affect postural steadiness in either young or older women. PMID- 16698286 TI - Cortical motor areas are activated early in a characteristic sequence during post movement processing. AB - During motor learning in goal-directed reactions, a specific movement has to be associated with feedback about the movement's success. Such feedback often follows when the movement is already over. We investigated the time-course of post-movement cortical motor processing by high-resolution analysis of lateralized post-movement potentials in forewarned and simple reaction time tasks. In both paradigms we could separate a post-movement component (motor postimperative negative variation-mPINV) peaking about 500 ms after the button press (confirmed by electromyogram and accelerometer). mPINV could not be sufficiently explained by motor cortex activity related to EMG output and/or by sensory feedback. mPINV was enhanced by long intertrial intervals and its lateralization changed with response movement side. Its scalp potential distribution resembled (pre-)motor cortex activity during preceding movement stages and differed from the frontal motor potential peak (proprioceptive and somatosensory reafferent feedback); suggesting post-movement activation of pre /primary motor cortex. Dipole source analysis yielded a single radial source near premotor cortex which explained lateralized mPINV almost completely. mPINV was present in simple reaction time tasks, indicating that mPINV is an independent component and does not represent delayed resolution of pre-movement negativity. An equivalent of "classical" PINV (cPINV) occurred later over prefrontal and anterior temporal sites in simple and forewarned reaction time tasks. Our results suggest that high-resolution analysis of lateralized movement-related potentials allows to image post-movement motor cortex activity and might provide insights into basic mechanisms of motor learning: A characteristic sequence might involve motor cortex activation (mPINV) before "higher order associative areas" come into play (cPINV). PMID- 16698287 TI - Brain and behavioral indices of retrieval mode. AB - In recent event-related potential (ERP) studies of episodic retrieval, ERPs have been acquired in tasks where participants have been cued trial-by-trial to prepare either to make episodic or semantic retrieval judgments. ERPs elicited during this preparatory cue period and separated according to retrieval task have diverged at right frontal scalp electrodes, with a relatively greater positivity associated with preparation for episodic rather than for semantic retrieval. Importantly, this pattern of differences has been observed only on 'stay' trials: those trials where the participant was cued to prepare for the same retrieval task on the previous trial. These findings have provided the basis for the proposal that the ERP modulations index processes that support the adoption or configuration of retrieval mode - a tonic process that can be sustained while recovery of episodic information is required and which facilitates the retrieval process. In these studies, however, the preparatory period on each trial was no more than 2000 ms, raising the possibility that, with more time available, neural correlates of these preparatory processes would not be restricted to stay trials. In this experiment, participants were cued trial-by-trial to complete either an episodic or a semantic retrieval task, and the preparatory period was greater than 4000 ms on the majority of trials. In keeping with previous findings, the ERPs elicited by these two cue types diverged principally on stay trials at right frontal electrode locations, suggesting that time to prepare is not the primary determinant of the onset of task-specific preparatory retrieval processing. In an important addition to previous findings, moreover, the accuracy of episodic memory judgments increased with the number of successive trials of the same task that participants completed, a finding consistent with the view that adopting a retrieval mode successfully can influence the accuracy of episodic memory judgments. PMID- 16698288 TI - Molecular clocks and the origin(s) of modern amphibians. PMID- 16698289 TI - Phylogenetic relationships and divergence times among New World monkeys (Platyrrhini, Primates). AB - Orthologous sequences of six nuclear genes were obtained for all recognized genera of New World monkeys (Primates: Platyrrhini) and outgroups to evaluate the phylogenetic relationships and to estimate divergence times. Phylogenetic relationships were reconstructed by maximum parsimony, maximum likelihood, and Bayesian approaches. All methods resolved with 100% branch support genus-level relationships, except for the grouping of Aotus as a sister taxa of Cebus and Saimiri, which was supported by low bootstrap percentages and posterior probability. All approaches depict three monophyletic New World monkey families: Atelidae, Cebidae, and Pitheciidae; also within each family, all approaches depict the same branching topology. However, the approaches differ in depicting the relationships of the three families to one another. Maximum parsimony depicts the Atelidae and Cebidae as sister families next joined by the Pitheciidae. Conversely, likelihood and Bayesian phylogenetic trees group families Atelidae and Pitheciidae together to the exclusion of Cebidae. Divergence time estimations using both local molecular clock and Bayesian approaches suggest the families diverged from one another over a short period of geological time in the late Oligocene-early Miocene. PMID- 16698290 TI - Inhibition of caspase-9 reduces chondrocyte apoptosis and proteoglycan loss following mechanical trauma. AB - OBJECTIVE: Chondrocyte death, a notable feature of osteoarthritis, may play a role in the initiation of cartilage degeneration. The present study was aimed at uncovering the nature and involvement of cell death in the initiation of cartilage degeneration induced by mechanical trauma. METHODS: Articular cartilage discs obtained from healthy skeletally mature horses were subjected to a single impact load (500 g from 50 mm) using a simple drop-tower device and cultured in vitro for 48 h. Chondrocyte death was examined using two independent methods: transmission electron microscopy and caspase-3 activity assay. To elucidate the signalling mechanisms involved in impact-induced cell death measured by terminal deoxynucleotidyl transferase-deoxyuridine triphosphate (dUTP) nick-end labelling (TUNEL), cartilage discs were incubated with specific caspase-3, -8 and -9 inhibitors prior to impact. Additionally, weight gain and glycosaminoglycan (GAG) release, markers of cartilage degeneration were monitored. RESULTS: After 48 h, ultrastructural evidence of apoptosis was observed. Caspase-3 was activated after 12h of culture post-impact. When quantified by TUNEL, impact trauma induced death in 52.6% of superficial chondrocytes after 48 h in culture, compared to 4.2% in unimpacted controls. Specific caspases-3 and -9 inhibitors significantly reduced impact-induced apoptosis to 24.3% and 14.7%, respectively. Caspase-8 inhibition had no effect on chondrocyte death (60.3%). Impact-induced GAG release into the medium was significantly reduced by inhibition of cell death, but weight gain remained unaffected by caspase inhibition. CONCLUSION: These results suggest that impact trauma-induced chondrocyte death is predominantly due to caspase-9 dependent apoptosis and is linked to cartilage degeneration. PMID- 16698291 TI - Calcitonin is involved in cartilage homeostasis: is calcitonin a treatment for OA? AB - OBJECTIVE: Osteoarthritis (OA) is the most common form of degenerative joint diseases and a major cause of disability and impaired quality of life in the elderly. Recent observations suggest that calcitonin may act on both osteoclasts and chondrocytes. The present review was sought to summarize emerging observations from the molecular level to the preliminary clinical findings of possible chondroprotective effects of calcitonin. METHOD: This review summarizes peer-reviewed articles found using pre-defined search criteria and published in the PubMed database before January 2006. In addition, abstracts from the OsteoArthritis Research Society International (OARSI) conferences in the time period 2000-2005 have been included in the search. RESULTS: Ample evidence for the effect of calcitonin on bone resorption was found. Support for direct effects of calcitonin on chondrocytes on matrix synthesis and inhibition of cartilage degradation have been published. In addition, clinical evidence for the effect of calcitonin on cartilage degradation is emerging. CONCLUSION: Several independent lines of evidence suggest a direct chondroprotective effect of calcitonin in addition to the well-established effect on bone resorption. Given the currently limited availability of chondroprotective agents, much expectation regards the ongoing clinical assessment of calcitonin therapy for the prevention and treatment of OA. PMID- 16698292 TI - Isolation from canned foods of a novel Thermoanaerobacter species phylogenetically related to Thermoanaerobacter mathranii (Larsen 1997): emendation of the species description and proposal of Thermoanaerobacter mathranii subsp. Alimentarius subsp. Nov. AB - Several anaerobic, thermophilic, Gram-positive bacteria were isolated from dairy products and canned meats. While some isolates were identified as Thermoanaerobacter thermohydrosulfuricus, comparisons of 16S rDNA genes indicated that others were phylogenetically closely related to Thermoanaerobacter mathranii, and more distantly related to Thermoanaerobacter thermocopriae and Thermoanaerobacter italicus. Biochemical characteristics, phylogenetic analysis, G+C content, and DNA-DNA hybridization experiments demonstrated that the strains AIP 504.99, AIP 505.99T and AIP 431.03, notwithstanding their high sequence similarities differ from T. mathranii and represent a novel T. mathranii subspecies for which the name T. mathranii subsp. alimentarius is proposed. The type strain is strain AIP 505.99T = CIP 108280T = CCUG 49566T. Emendation of the species description for T. mathranii is proposed to include this subspecies. PMID- 16698293 TI - Investigating RGS proteins in yeast. AB - Regulator of G protein signalling (RGS) proteins are vital in the adaptation of cells to stimulation via G protein-coupled receptors. Yeast have been integral in elucidating the important role that RGS proteins play within cellular processes. In addition to extensive characterisation of the endogenous RGS proteins, these organisms have enabled the identification and analysis of numerous mammalian homologues. The simplicity and plasticity of the yeast pheromone-response pathway has facilitated studies which would have been impossible in mammalian systems and it is certain that yeast will continue to have a great impact on this field of research in the future. PMID- 16698294 TI - Gene expression profiling of the nitrogen starvation stress response in the mycorrhizal ascomycete Tuber borchii. AB - The focus of this work is on the nitrogen starvation stress responses operating in a plant symbiotic fungus. A cDNA array profiling analysis was conducted on N limited mycelia of the mycorrhizal ascomycete Tuber borchii. Fifty-one unique transcripts, out of 2062 redundant arrayed cDNAs, were differentially expressed by at least 1.5-fold in response to N deprivation. Only two N assimilation components-a nitrate transporter and a high-affinity ammonium transporter-were found among differentially expressed genes. All the other N status responsive genes code for as yet unidentified hypothetical proteins or components not directly involved in N assimilation or metabolism, especially carbohydrate binding proteins and oligosaccharide as well as lipid modifying enzymes. A subset of cDNA array data were confirmed and extended by Northern blot analysis, which showed that most of the latter components respond not only to nitrogen, but also to carbon source depletion. PMID- 16698295 TI - Transcription analysis using high-density micro-arrays of Aspergillus nidulans wild-type and creA mutant during growth on glucose or ethanol. AB - Here, we describe how the recently published Aspergillus nidulans genome sequence [Galagan, J.E., Calvo, S.E., Cuomo, C., Li-Jun, M., Wortman, J.R., et al., 2005. Sequencing of Aspergillus nidulans and comparative analysis with A. fumigatus and A. oryzae. Nature 438 (7071), 1105-1115] was used to design a high-density oligo array with probes for 3,278 selected genes using the Febit Geniom One array system. For this purpose, the program OligoWiz II was used to design 24,125 probes to cover the 3,278 selected genes. Subsequently, the Febit system was used to investigate carbon catabolite repression by comparing the gene expression of a creA deleted mutant strain with a reference strain grown either with glucose or ethanol as the sole carbon source. In order to identify co-regulated genes and genes influenced by either the carbon source or CreA, the most significantly regulated genes (pDPAn-3> or =EPA>AA approximately 20:3(n-6)> or =22:4(n-6). One step desaturation, C2 elongation or C2 cleavage within the n-6 or n-3 fatty acid group was probably very active in L929 cells, because AA, synthesized from 20:3(n-6) or 22:4(n-6), and C22 fatty acids, synthesized from AA or EPA, were preferentially retained in cellular phospholipids. These observations suggested that attenuation of TNF induced necrosis by the supplementation of various C20 or C22 polyunsaturated fatty acids is mainly attributable to the enrichment of three kinds of polyunsaturated fatty acids, i.e., DHA, DPAn-3 or AA, in phospholipids. Among these fatty acids, DHA was the most effective in the reduction of L929 necrosis as observed in the case of U937 apoptosis. This suggests that DHA-enriched membranes can protect cell against TNF irrespective of death modes and that membranous DHA may abrogate the death signaling common to necrosis and apoptosis. PMID- 16698314 TI - Modification of low-density lipoprotein by myeloperoxidase-derived oxidants and reagent hypochlorous acid. AB - Substantial evidence supports the notion that oxidative processes contribute to the pathogenesis of atherosclerosis and coronary heart disease. The nature of the oxidants that give rise to the elevated levels of oxidised lipids and proteins, and decreased levels of antioxidants, detected in human atherosclerotic lesions are, however, unclear, with multiple species having been invoked. Over the last few years, considerable data have been obtained in support of the hypothesis that oxidants generated by the heme enzyme myeloperoxidase play a key role in oxidation reactions in the artery wall. In this article, the evidence for a role of myeloperoxidase, and oxidants generated therefrom, in the modification of low density lipoprotein, the major source of lipids in atherosclerotic lesions, is reviewed. Particular emphasis is placed on the reactions of the reactive species generated by this enzyme, the mechanisms and sites of damage, the role of modification of the different components of low-density lipoprotein, and the biological consequences of such oxidation on cell types present in the artery wall and in the circulation, respectively. PMID- 16698317 TI - Long-term changes in leisure time walking, moderate and vigorous exercise: were they influenced by the National Physical Activity Guidelines? AB - Data from the Australian Bureau of Statistics three cross-sectional National Health Surveys, conducted in 1989-1990, 1995-1996 and 2000, were used to examine long-term population responses to the 1996 physical activity (PA) recommendations. Past 2-week recall of the number of times and total minutes spent walking for recreation or sport, and doing moderate and vigorous exercise, were averaged for 1 week to create a measure of 'sufficiently active': >or=5 weekly sessions of >or=150 min/week of at least moderate-intensity activity. With the exception of adults aged>60 years, from 1989 to 2000, there were increases in the prevalence of walking (+8%) and in moderate-intensity exercise (+4%), resulting in a significant decline (-6%) in the proportion of those reporting no leisure-time exercise. Participation in vigorous exercise remained unchanged. There was a 'population shift' in median minutes and sessions of walking from 1995 to 2000, but the prevalence of regular walking (>or=5 times and >or=150 min/week) remained unchanged. A small but significant increase (+2%) in the proportion of adults who met the PA recommendations was noted mainly for men under 60 years and for women aged 45-59 years. The proportion meeting the PA recommendations declined significantly in the most socioeconomically disadvantaged area. These data provide evidence contrary to the observed declines in PA documented by the Active Australia surveillance in the late 1990s. The population may have responded to the 1996 concept of moderate-intensity exercise mainly by walking, but sustainable promotional efforts will be required to build on these gains in the forthcoming decades. PMID- 16698318 TI - Evaluation of Epsilon amino-caproic acid (EACA) and autologous blood as blood conservation strategies in patients undergoing cardiac surgery. AB - BACKGROUND: To evaluate the effects of autologous blood and Epsilon amino-caproic acid on intra-operative and post-operative blood loss and homologous blood product requirements in patients undergoing cardiac surgery. METHODS: Patients were randomly allocated to two groups of 30 each. In the Epsilon amino-caproic acid (EACA) group, the drug was administered in a loading dose of 100 mg/kg before skin incision followed by an infusion of 1/5 th the loading dose hourly and terminated 3 h after heparin neutralization. In the autologous transfusion (AT) group, 10% of the calculated whole blood volume was collected intra operatively before cardiopulmonary bypass and re-infused after its termination. RESULTS: Haemoglobin values were comparable pre-operatively, on cardiopulmonary bypass, off cardiopulmonary bypass and post-operatively on day two in both groups. Intra-operative blood loss was not significantly different (643.3+/ 129.14 ml in group EACA versus 710+/-145.5 ml in group AT, p = 0.66). Although the chest drainage was more in group AT during 0-3 h (71.3+/-54.3 ml versus 112.6+/-79.3.6 ml, p = 0.006) it was comparable amongst in the first 24 h (231.1+/-98.3 ml in group AT versus 235+/-101.4 ml in group EACA, p = 0.88). Homologous blood product requirements were similar in both groups. CONCLUSION: Autologous blood is as efficacious as Epsilon amino-caproic acid for blood conservation in cardiac surgery. PMID- 16698319 TI - Production of interferon-gamma by myeloid cells--fact or fancy? AB - Myeloid cells are usually viewed as targets rather than producers of interferon (IFN)-gamma. However, since 1993 several studies have suggested that macrophages and dendritic cells can also secrete IFN-gamma in response to various stimuli, which led to the idea of autocrine myeloid-cell activation in innate immunity. Here, we review this concept in the light of recent findings that illustrate the necessity of specific cell markers, cell purity, sensitive single-cell cytokine detection methods, and of further insights into the developmental origin of myeloid and lymphoid cell lineages. We also discuss the possible function of intracellular IFN-gamma in macrophages and the evidence for a protective role of myeloid IFN-gamma in vivo. PMID- 16698320 TI - Counselling on lifestyle factors in hypertension care after training on the stages of change model. AB - BACKGROUND: In assisting the nurse's counselling on lifestyle changes in hypertension care a behaviour model can be used. AIM: To analyse the effects of nurses' training on the use of the stages of change model when counselling hypertensive patients to perform lifestyle changes. METHODS: As part of a randomised, controlled trial, 19 nurses belonging to the intervention group took part in video-recorded consultation training with simulated patients. To evaluate the training, the nurses audio-recorded their consultations with two patients before and after the intervention. Analysis focused on the areas of non pharmacological treatment and the nurses' attention to the patients' readiness for change. RESULTS: Patient participation in the consultations increased after the training. The importance of non-pharmacological treatment was mentioned more frequently for all areas of lifestyle behaviour, exercise, smoking, alcohol consumption, food and stress, and the nurses acquired a more distinct structure for their consultations. The mean length of the recorded consultations increased from 18 min to 20.5 min. All the criteria for fulfillment of attention to patient's readiness to change were met in nine consultations before the training and in seven after it. After the training, attention was paid to support more frequently than before in the action and maintenance stages and a great deal of information was provided. PMID- 16698321 TI - Mucopolysaccharidosis I cats mount a cytotoxic T lymphocyte response after neonatal gene therapy that can be blocked with CTLA4-Ig. AB - Although gene therapy has reduced manifestations of genetic diseases, immune responses can abrogate the effect. One approach to inducing tolerance is to perform gene transfer in newborns when the immune system is immature. We demonstrate here that the dose of retroviral vector (RV) is important in mice, as mucopolysaccharidosis I (MPS I) mice that received neonatal intravenous gene therapy with a high dose of a canine alpha-L-iduronidase (cIDUA)-expressing RV had stable expression, while those that received a low dose did not. It was unclear, however, if neonatal transfer with any dose could induce tolerance in large animals. Therefore, newborn MPS I cats were injected intravenously with the RV expressing cIDUA. Although this resulted in high serum IDUA activity due to secretion by transduced cells, expression fell due to a CTL response. Cats that transiently received the immunosuppressive agent CTLA4-Ig did not develop a CTL response. In contrast, MPS I dogs, which can respond immunologically to canine IDUA, had stable serum IDUA activity after neonatal gene therapy. We conclude that cats, but not dogs, mount a potent CTL response to canine IDUA after neonatal gene therapy, which can be prevented with transient CTLA4-Ig. PMID- 16698322 TI - Labeling and intracellular tracking of functionally active plasmid DNA with semiconductor quantum dots. AB - Semiconductor nanocrystal quantum dots (QDs) allow long-term imaging in the cellular environment with high photostability. QD biolabeling techniques have previously been developed for tagging proteins and peptides as well as oligonucleotides. In this contribution, QD-decorated plasmid DNA was utilized for the first time for long-term intracellular and intranuclear tracking studies. Conjugation of plasmid DNA with phospholipid-coated QDs was accomplished using a peptide nucleic acid (PNA)-N-succinimidyl-3-(2-pyridylthio) propionate linker. Gel electrophoresis and confocal and atomic force microscopy (AFM) were used to confirm the structure of QD-DNA conjugates. AFM imaging also revealed that multiple QDs were attached in a cluster at the PNA-reactive site of the plasmid DNA. These QD-DNA conjugates were capable of expressing the reporter protein, enhanced green fluorescent protein, following transfection in Chinese hamster ovary (CHO-K1) cells with an efficiency of ca. 62%, which was comparable to the control (unconjugated) plasmid DNA. PMID- 16698323 TI - Hot water epilepsy: clinical and electroencephalographic features of 25 cases. AB - The aim of this study was to analyze the clinical and electroencephalographic findings for 25 patients with hot water epilepsy. Personal and family history, neurological state, age at onset and types of seizures, provoking factors, bathing habits, EEGs, and neuroimages were all reviewed. Age at onset of seizures ranged from 6 months to 37 years. Twenty patients had complex partial seizures. Eight patients had spontaneous seizures as well; one was a typical case of idiopathic photosensitive occipital lobe epilepsy. One patient described an episode interpreted as nonconvulsive status. In our study group, the pouring of the water over the head and the temperature of the water were the most common triggering factors. A special kind of soap and entry of water into the mouth were determined to be unusual triggering factors. Interictal EEGs revealed epileptogenic abnormalities located over the temporal regions in nine patients. Seventeen patients underwent neuroimaging, mostly cranial magnetic resonance imaging. One had right mesial temporal sclerosis and one cortical atrophy; the others had normal findings. Sixteen patients received antiepileptic drugs, mainly carbamazepine, and remained seizure-free. The high rate of epileptogenic abnormalities localized in the temporal region and the complex partial seizures observed in most of our patients indicate the considerable role of the temporal lobe in hot water epilepsy. PMID- 16698324 TI - Gene therapy of the ischemic lower limb--Therapeutic angiogenesis. AB - The limitations of surgical revascularisation and pharmacological treatment in peripheral arterial occlusive disease (PAOD) are well recognized. Therapeutic options for critical leg ischemia are consequently limited to percutaneous transluminal angioplasty (PTA) or surgical revascularisation. Unfortunately, many patients with critical leg ischemia are poor candidates for either procedure. Therapeutic angiogenesis is a novel promising tool to treat these patients. Experimental and clinical and trials of gene transfer for therapeutic angiogenesis have already shown some clinical efficacy. This review is focused on gene transfer techniques in preclinical and clinical therapeutic angiogenesis, angiogenic growth factors, vectors, delivery methods and routes. The results of clinical and experimental studies, safety and side effects of gene therapy, and the perspectives of future research are also discussed. PMID- 16698325 TI - Evaluation of cytotoxic effect of photodynamic therapy in combination with electroporation in vitro. AB - Under the influence of electric pulses cells undergo membrane electroporation (EP), which results in increased permeability of the membrane to exogenous compounds. EP is applied in oncology as a method to enhance delivery of anticancer drugs. For that reason it was essential to combine photodynamic tumor therapy (PDT)--the cancer treatment method based on the use of photosensitizers that localize selectively in malignant tumors and become cytotoxic when exposed to light, and EP, with the aim to enhance the delivery of photosensitizers into the tumor and therefore to increase the efficacy of PDT. Thus, the aim of study was to evaluate the cytotoxic effect of PDT in combination with EP. A Chinese hamster lung fibroblast cell line (DC-3F) was used. The cells were affected by photosensitizers chlorin e(6) (C e(6)) at the dose of 10 mug/ml and aluminium phthalocyanine tetrasulfonate (AlPcS4) at the dose of 50 microg/ml. Immediately after adding of photosensitizers the cells were electroporated with 8 electric pulses at 1200 V/cm intensity, 0.1 ms duration, 1 Hz frequency. Then, after 20 min of incubation the cells were irradiated using a light source--a visible light passing through a filter (KC 14, emitted light from 660 nm). The fluence rate at the level of the cells was 3 mW/m(2). Cytotoxic effect on cells viability was evaluated using MTT assay. Our in vitro data showed that the cytotoxicity of PDT in combination with EP increases fourfold on the average. Based on the results we suggest that EP could enhance the effect of PDT. PMID- 16698326 TI - Selective response of dopamine in the presence of ascorbic acid on L-cysteine self-assembled gold electrode. AB - A L-cysteine (L-Cys) self-assembled modified gold electrode was used to detect dopamine (DA) by chronoamperometric method (CE) in the presence of ascorbic acid (AA). The defective limit is 2.0 x 10(-8) mol L(-1) (S/N=3). The proposed method was applied to detect DA in the samples with satisfied result. PMID- 16698327 TI - A new simple HPLC method for measuring mitotane and its two principal metabolites Tests in animals and mitotane-treated patients. AB - A new C18 reversed-phase column and UV HPLC method for the detection of mitotane, its principal metabolites, dichlorodiphenylacetate and dichlrodiphenylethene, and its precursor DDT is described. In this article mitotane, dichlorodiphenylacetate, and dichlrodiphenylethene concentrations in organs of rats fed on a mitotane diet, and the effects of erythromycin and grapefruit juice as cytochrome P450 common inhibitors are presented. Tissue accumulation of mitotane and dichlrodiphenylethene, the acquired ability to eliminate dichlorodiphenylacetate, and inhibition of beta-hydroxylation by both inhibitors are illustrated here. Blood samples from mitotane-treated patients revealed two correlations: plasma mitotane/dichlrodiphenylethene and plasma mitotane/red cell mitotane. PMID- 16698328 TI - Screening of blood donors for tissue transglutaminase antibodies in the Ryazan area (Russia). PMID- 16698329 TI - Introduction: there is an important need for increased understanding of the pathophysiology of heart failure postinfarction, new treatment strategies, earlier implementation of effective strategies, and more effective use of existing therapies. PMID- 16698330 TI - Cardiovascular effects of aldosterone and post-acute myocardial infarction pathophysiology. AB - Aldosterone is an important mediator of the renin-angiotensin-aldosterone system (RAAS) that plays a major role in the pathophysiology of cardiovascular disease as well as regulation of extracellular fluid volume and potassium. In experimental models, aldosterone has been shown to promote endothelial dysfunction; induce vascular inflammation, myocardial ischemia, and necrosis; increase collagen synthesis in cardiac fibroblasts; contribute to plasminogen activator inhibitor-1 regulation; decrease baroreceptor sensitivity and reflex function; block myocardial uptake of norepinephrine; increase oxidative stress; and stimulate cardiomyocyte apoptosis. A review of animal and human studies with aldosterone blockers reveals improvement in, and in some cases complete reversal of, these pathophysiologic effects of aldosterone on the cardiovascular system. PMID- 16698331 TI - Epidemiology of heart failure and left ventricular systolic dysfunction after acute myocardial infarction: prevalence, clinical characteristics, and prognostic importance. AB - The development of heart failure and/or left ventricular systolic dysfunction (LVSD) in the setting of acute myocardial infarction (AMI) results in significant risk far above that of AMI independently. In patients admitted to the hospital for AMI, concomitant heart failure and/or LVSD on hospital admission or development of either or both of these conditions during admission are among the strongest predictors of inhospital death and are associated with significant increases in inhospital, 30-day, and long-term mortality and rehospitalization rates. Given the high risks in this population, aggressive treatment, comprising early initiation and sustained use of evidence-based treatments, is essential for improving prognosis. PMID- 16698332 TI - Improving outcomes in post-acute myocardial infarction heart failure: incorporation of aldosterone blockade into combination therapy to optimize neurohormonal blockade. AB - Although angiotensin-converting enzyme (ACE) inhibitors, angiotensin II receptor blockers (ARBs), and beta-blockers have been proved to reduce mortality in patients with heart failure post-acute myocardial infarction (AMI), studies show that these agents are consistently underused in this population. Further, morbidity and mortality remain high even when standard-of-care therapies are applied. Thus, new strategies have been sought to better counteract the maladaptive effects of neurohormonal stimulation in post-AMI heart failure. The Eplerenone Post-Acute Myocardial Infarction Heart Failure Efficacy and Survival Study (EPHESUS) demonstrated that the selective aldosterone blocker eplerenone, when used in addition to standard therapy, results in significant incremental improvements in survival and morbidity and is safe and well tolerated in this setting. Based on this, major therapeutic guidelines in the United States and Europe now strongly recommend that all eligible patients with concomitant heart failure post-AMI be treated with an aldosterone blocker in addition to an ACE inhibitor (or an ARB) and a beta-blocker. To achieve needed improvements in outcomes in this population, early and consistent initiation of these evidence based, guideline-recommended therapies in all eligible patients is crucial. PMID- 16698333 TI - Role of aldosterone blockade for treatment of heart failure and post-acute myocardial infarction. AB - Heart failure continues to significantly impact morbidity and mortality after acute myocardial infarction (AMI), especially when associated with left ventricular systolic dysfunction (LVSD). Therefore, routine use of optimal therapy for patients with heart failure after AMI is needed in the future. Data from the Eplerenone Post-Acute Myocardial Infarction Heart Failure Efficacy and Survival Study (EPHESUS) has demonstrated that addition of eplerenone to standard therapy in patients with LVSD and clinical heart failure post-AMI significantly improves outcomes in these patients, providing the basis for acceptance of aldosterone blockade as an integral component of therapy. However, further studies are needed to more broadly explore the usefulness of aldosterone blockade in heart failure therapy. As research clarifies the many complex pathophysiologic processes involved in post-AMI remodeling, treatment should continue to move beyond symptom-targeted therapies to disease- and mechanism-targeted therapies that will further improve outcomes in the setting of heart failure post-AMI and beyond. PMID- 16698334 TI - Perioperative nursing. PMID- 16698335 TI - Preoperative nursing assessment of the surgical patient. AB - Thorough assessment of the surgical patient begins in the preoperative phase and extends throughout the perioperative experience. Patient-centered interviews gather critical data that contribute to a successful experience and a focused plan of care. Information collected, documented, and conveyed to the surgeon or physician assists in appropriate medical decision making. This article focuses on perioperative nursing assessment tools and discusses issues pertinent to achieving safe delivery of care. A broad overview touches on key topics that deserve exploration and evaluation. This article will hopefully serve as a useful tool that helps direct the decision-making process. PMID- 16698336 TI - Prevention, recognition, and management of nursing complications in the intraoperative and postoperative surgical patient. AB - A review of the literature focusing on postoperative complications reveals that the best available tools to the medical and surgical teams are recognition and prevention. This article highlights the more common postsurgical adverse events and discusses methods for preventing and treating these occurrences. PMID- 16698337 TI - Positioning impact on the surgical patient. AB - The operating room is unknown territory to most health care providers. It frequently brings up thoughts of blood, strange smells, and cold temperatures. Many nursing programs have scheduled little, if any, time in this environment for students. As a result, few nurses who practice outside of this specialized area understand the patient care events that occur in the operating room. Those who have selected the operating room for their work environment know that it is a somewhat isolated period in the perioperative experience. This article provides insight into this area of patient care and a greater understanding of how patients are positioned, the physiologic impact of these positions, and some consequences that may impact the postoperative care of these patients. PMID- 16698338 TI - Comparison of operating room lasers: uses, hazards, guidelines. AB - Although the discovery of laser light no longer can be termed recent, it took the medical community a long time to use its technology. Everyday, advances are being made in laser technology, and new applications are being discovered for this modality. Because lasers allow for the capture, control, and manipulation of energy, it is important for nurses to understand the basic biophysics of lasers. As lasers are being used in most surgical settings and specialties, it is important for the perioperative nurse to have a basic knowledge of each wave length, type of laser, indications for its use, and safety considerations. PMID- 16698339 TI - Laparoscopy: risks, benefits and complications. AB - This article discusses the benefits and risks of laparoscopy. Also discussed are complications of laparoscopy and methods to avoid or treat these adversities. PMID- 16698340 TI - How religion, language and ethnicity impact perioperative nursing care. AB - Religion, language, and ethnicity play important roles in the perioperative arena. This article highlights some of the challenges that religion, language, and ethnicity can present and offers strategies for making the experience as positive as possible for all patients. PMID- 16698341 TI - Bariatric surgery risks, benefits, and care of the morbidly obese. AB - The American Society for Bariatric Surgery defines morbid obesity as a lifelong, progressive, life-threatening, genetically-related, costly, multi-factorial disease of excess fat storage with multiple comorbidities. Obesity satisfies the definition of morbid obesity when it reaches the point of significant risk for obesity-related comorbidities. These significant comorbidities often result in either significant physical disability or even death. Obesity results from excessive accumulation of fat that exceeds the body skeletal and physical standards. Morbid obesity is defined as being at least 100 lb heavier than ideal body weight, or a body mass index (BMI) of 40. BMI is calculated as weight in kilograms divided by the height in meters squared. This article focuses on the causes, treatment, and perioperative nursing care of patients who are morbidly obese. PMID- 16698342 TI - Unique concerns of the pediatric surgical patient: pre-, intra-, and postoperatively. AB - This article focuses on the unique concerns of pediatric surgical patients and emphasizes preparation of the family as an integrated unit. The statement from older standards of the American Nurses' Association that "children have the right to be treated with dignity and respect" remains a valid challenge for nurses caring for these patients. PMID- 16698343 TI - Management of the special needs of the pregnant surgical patient. AB - This article summarizes the nursing care that should be provided to the pregnant patient during surgery and gives a brief discussion of the physiology of pregnancy as it relates to surgery and anesthesia. An overview of anesthesia considerations is presented and the nursing process using the guidelines for pregnant patients from the Association of Perioperative Registered Nurses is discussed, followed by nursing considerations for the most common surgical procedures that pregnant women undergo. PMID- 16698344 TI - Nursing considerations in the geriatric surgical patient: the perioperative continuum of care. AB - Although aging is a natural process, caring for an older person is not the same as caring for a middle-aged adult person. The perioperative and perianesthesia nurses need to be competent in geriatric nursing care and possess specialized knowledge and skills related to a myriad of geriatric issues. This article focuses on the special needs of the elderly population and how the perioperative nursing team may address their care best. PMID- 16698345 TI - Current trends in molecular classification of adult renal tumors. PMID- 16698346 TI - Management of chronic prostatitis/chronic pelvic pain syndrome: an evidence-based approach. PMID- 16698347 TI - What is the most cost-effective treatment for 1 to 2-cm bulbar urethral strictures: societal approach using decision analysis. AB - OBJECTIVES: Direct vision internal urethrotomy (DVIU) and urethroplasty are the primary methods of managing urethral stricture disease. Using decision analysis, we determine the cost-effectiveness of different management strategies for short, bulbar urethral strictures 1 to 2 cm in length. METHODS: A decision tree was constructed, with the number of planned possible DVIUs before attempting urethroplasty defined for each primary branch point. Success rates were obtained from published reports. Costs were estimated from a societal perspective and included the costs of the procedures and office visits and lost wages from convalescence. Sensitivity analyses were conducted, varying the success rates of the procedures and cost estimates. RESULTS: The most cost-effective approach was one DVIU before urethroplasty. The incremental cost of performing a second DVIU before attempting urethroplasty was $141,962 for each additional successfully voiding patient. In the sensitivity analysis, urethroplasty as the primary therapy was cost-effective only when the expected success rate of the first DVIU was less than 35%. CONCLUSIONS: The most cost-effective strategy for the management of short, bulbar urethral strictures is to reserve urethroplasty for patients in whom a single endoscopic attempt fails. For longer strictures for which the success rate of DVIU is expected to be less than 35%, urethroplasty as primary therapy is cost-effective. Future prospective, multicenter studies of DVIU and urethroplasty outcomes would help enhance the accuracy of our model. PMID- 16698348 TI - Long-term efficacy of holmium laser endoureterotomy for benign ureteral strictures. AB - OBJECTIVES: To determine the long-term clinical and radiographic success of holmium laser endoureterotomy for nonobliterative benign ureteral strictures. METHODS: A total of 19 patients (12 women and 7 men, mean age 47.5 years) underwent holmium laser endoureterotomy for iatrogenic ureteral strictures (seven proximal, seven mid, and five distal) using semirigid ureteroscopy and a 360 microm fiber at 1 J and 10 Hz. RESULTS: Success was strictly defined as both relief of symptoms and radiographic resolution of obstruction by intravenous pyelography or diuretic renography, or both. With a median follow-up of 3.0 years, success was achieved in 13 (68.4%) of 19 patients. CONCLUSIONS: Our results have shown that holmium laser endoureterotomy is associated with a long term success rate equivalent to, or better than, other currently available minimally invasive treatment options. Also, failure was uniformly evident within the first 3 months after treatment. PMID- 16698349 TI - Comparison of mechanical and in vivo performance of new and reprocessed harmonic scalpels. AB - OBJECTIVES: To compare the mechanical and clinical performance of new and reprocessed harmonic scalpels (HS). METHODS: A total of 89 reprocessed and 90 new HS (laparoscopic coagulating shears with a curved blade and 5-mm-diameter shaft) were subjected to visual inspection, destructive testing, and nondestructive mechanical testing. Subsequently, new HS, randomly selected reprocessed HS, and selected reprocessed HS with known abnormalities were graded on clinical performance by 14 surgeons in a porcine model. RESULTS: Visual inspection of the HS discriminated between the new and reprocessed instruments in 11 of the 12 visual criteria (P < or = 0.02). In vitro mechanical testing revealed greater clamp arm dislodge forces for the new HS (P < 0.01) and greater midshaft temperatures for reprocessed HS (P < 0.01). Overall, 65 (73%) of 89 reprocessed and 7 (7.8%) of 90 new HS had gross abnormalities noted on inspection and mechanical evaluation (P < 0.01). The surgeons' evaluation of the instruments during in vivo testing demonstrated that the new instruments manifested significantly greater hemostatic control compared with randomly selected reprocessed HS (P = 0.01) and were significantly better for tissue sticking (P = 0.01), tissue transection (P = 0.02), tissue dissection (P < 0.01), grasping (P < 0.01), and hemostatic control (P = 0.04) compared with reprocessed HS with obvious defects. CONCLUSIONS: In vitro and in vivo data demonstrated significantly greater performance for new HS compared with reprocessed HS. Mechanical testing of reprocessed HS could not adequately distinguish which HS would result in decreased performance. These findings raise important issues of clinical safety in the contemporary quest for cost-effectiveness. PMID- 16698350 TI - Heat/burning sensation induced by topical application of capsaicin on perineal cutaneous area: new approach in diagnosis and treatment of chronic prostatitis/chronic pelvic pain syndrome? AB - OBJECTIVES: To investigate the feasibility, safety, and efficacy of perineal cutaneous application of capsaicin as a test for the diagnosis, as well as a potential therapeutic tool, in patients with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS). METHODS: We recruited 22 patients (mean age 34.5 years, range 19 to 56), who had been diagnosed with CP/CPPS according to the National Institutes of Health criteria, and 6 healthy control subjects. Both groups received a topical application of 5 mL capsaicin at a concentration of 10(-5) M to the perineal body skin. The patients were asked to mark on a visual analogue scale the intensity of any heat or burning sensation. All the patients had completed a National Institutes of Health Chronic Prostatitis Symptom Index before and 1 week after the test. The scores of the two groups were compared using the Mann-Whitney U test. RESULTS: The patients with CP/CPPS reported a heat/burning sensation intensity that was statistically greater than that of the healthy controls (7.5 versus 4.3, P <0.001) and a shorter time to heat sensation onset and maximal intensity (1.5 versus 3.4 minutes, P <0.001, and 2.5 versus 7 minutes, P <0.001, respectively). Of the 22 patients, 16 reported an improvement of symptoms after 7 days and the mean National Institutes of Health Chronic Prostatitis Symptom Index score decreased from 27 to 16 (P <0.01). CONCLUSIONS: We found a statistically significant difference in the pain visual analogue scale and interval between topical application and the onset of the heat/burning sensation between patients with CP/CPPS and healthy controls. The small sample size strongly suggests the need for additional larger and more controlled studies. PMID- 16698351 TI - Assessment of efficacy of transcutaneous electrical nerve stimulation for pain management during office-based flexible cystoscopy. AB - OBJECTIVES: To evaluate the efficacy of transcutaneous electrical nerve stimulation (TENS) for its effectiveness in eliminating or minimizing discomfort during office-based flexible cystoscopy. METHODS: A total of 148 patients were prospectively randomized into one of three groups: flexible cystoscopy with no analgesics, a placebo TENS, or an activated TENS. The patient data collected included patient parameters, number of previous flexible cystoscopies, visual analog pain scores during and after the procedure, surgeon's difficulty rating of procedure, and International Prostate Symptom Score before and 24 hours after the procedure. RESULTS: No statistically significant difference was found among the three groups regarding patient parameters. The visual analog pain scores were similar before and after the procedure. At 30 seconds, the mean visual analog scale score for the control group, placebo group, and TENS study group was 3.73, 3.65, and 3.52, respectively (control versus placebo, control versus active, and placebo versus active: P = 0.97, 0.29, and 0.53, respectively). At 1 and 5 minutes, the corresponding scores were 3.44, 4.37, and 3.50 (P = 0.88, P = 0.99, and P = 0.99) and 0.86, 1.23, and 0.88 (P = 0.97, P = 0.35, and P = 0.56), respectively. The surgeon's mean difficulty rating for the control procedures was 1.08 and for the placebo group was 2.30 (P = 0.02). CONCLUSIONS: All patients undergoing flexible cystoscopy in the office setting experienced discomfort. The TENS device provided no significant benefit for pain. The trend toward greater pain scores in the control group could be attributed to the greater degree of difficulty. PMID- 16698352 TI - Clinical sequelae of radiographic iceball involvement of collecting system during computed tomography-guided percutaneous renal tumor cryoablation. AB - OBJECTIVES: Percutaneous renal tumor cryoablation is being evaluated as a treatment option for small renal tumors. However, when tumors are located centrally, involvement of the collecting system by the radiographic iceball can occur. We reviewed our series of computed tomography (CT)-guided percutaneous renal tumor cryoablation to identify those cases in which there appeared to be involvement of the collecting system by the radiographic iceball and to determine any clinical sequelae of such involvement. METHODS: Retrospective review of the medical records identified 6 patients who had undergone CT-guided percutaneous renal tumor cryoablation with evidence of collecting system involvement. Measurements of the tumor size, size of the radiographic iceball, and the size of the immediate postprocedure "cryozone" (region of apparent treatment on contrast enhanced CT) were obtained from the preprocedure, intraprocedure, and immediate postprocedure CT scans. Follow-up imaging was obtained beginning at 3 to 6 months. RESULTS: Six patients were identified who met the inclusion criteria and had at least 3 months of documented follow-up. Despite the apparent involvement of the collecting system during the cryoablation procedure, no patient developed clinical signs or symptoms or radiographic evidence of a urine leak or fistula formation. Furthermore, no evidence of ureteral narrowing or stricture formation has been found to date, with a mean follow-up of 167.7 days (range 90 to 288). CONCLUSIONS: We observed no clinically appreciable urine leaks despite what appeared to be obvious involvement of the collecting system by the radiographic iceball. However, care should be exercised to avoid this insult when possible until additional research has confirmed its safety. PMID- 16698353 TI - Proposed classification of complications after live donor nephrectomy. AB - OBJECTIVES: A standardized classification for the potential complications of living donor nephrectomy is an essential step in establishing a construct for monitoring and reporting the outcomes of this procedure. It is also helpful in informing potential donors about the inherent risks of the donor operation as part of the informed choice process. METHODS: We reviewed 600 laparoscopic live donor nephrectomies performed at our center. A modification of the Clavien classification system describing procedure-related complications was developed and used to grade the severity of all complications. RESULTS: We observed 43 complications (7.2%) in our series of 600 patients. Grade 1 defines all events that, if left untreated, would have a spontaneous resolution or needed a simple bedside procedure (39.5%). Grade 2 complications differ from grade 1 in that they are potentially life-threatening and usually require some form of intervention, but do not result in ongoing disability. We subdivided grade 2 complications (55.8% in our study) into 2a, 2b, and 2c. The latter describes complications requiring open conversion of laparoscopic donor nephrectomy for patient treatment. Grade 3 complications are events with residual or lasting disability (4.7% in our review). Grade 4 events are those resulting in renal failure or death because of any complication (none occurred in our series). CONCLUSIONS: A graded classification scheme for reporting the complications of donor nephrectomy may be useful for maintaining registry information on donor outcomes and when informing potential donors about the risks and benefits of this procedure. PMID- 16698354 TI - Detection of carbonic anhydrase-9 gene expression in peripheral blood cells predicts risk of disease recurrence in patients with renal cortical tumors. AB - OBJECTIVES: To present extended follow-up on a cohort of patients with renal cortical tumors treated with partial or radical nephrectomy and preoperatively assess for carbonic anhydrase 9 tumor marker expression in the peripheral blood. METHODS: All patients were originally enrolled in an institutional review board approved study assessing the role of a reverse-transcriptase polymerase chain reaction peripheral blood assay designed to detect the tumor-specific gene carbonic anhydrase-9 (CA9). A total of 41 patients with renal cortical tumors confined to the kidney were enrolled at a single institution and assessed preoperatively with peripheral blood test for CA9 expression before undergoing partial or radical nephrectomy. A Kaplan-Meier estimated survival analysis and log-rank test were performed to determine whether detection of peripheral blood cells expressing the CA9 gene influences disease-free and disease-specific survival. RESULTS: The median follow-up was 4.3 years. The median age was 71 years. Of the 41 patients, 26 were men and 15 were women. The estimated 5-year disease-free survival for patients with detectible expression of the CA9 gene in the peripheral blood was 39.5% compared with 88.1% for patients without detection of the CA9 gene (P = 0.048). On bivariate analysis, disease-free survival correlated with histologic type, tumor diameter, and tumor grade. CONCLUSIONS: The expression of the tumor-specific marker CA9 in the peripheral blood is associated with decreased disease-free survival in patients with renal cortical tumors. This is the first study reporting on the prognostic value of this peripheral blood-based tumor marker for kidney tumors. PMID- 16698355 TI - uCyt+ test: alternative to cystoscopy for less-invasive follow-up of patients with low risk of urothelial carcinoma. AB - OBJECTIVES: To assess the utility of ImmunoCyt/uCyt+ in combination with cytology to reduce the number and cost of cystoscopies in the follow-up of patients with urothelial cancer of the bladder. METHODS: A total of 216 patients under follow up after transurethral resection of the bladder for urothelial cancer were enrolled in the present study. The mean follow-up time was 26 months (range 3 to 96). Patients were tested for bladder cancer with ImmunoCyt/uCyt+, urinary cytology, and cystoscopy every 3 months. All patients with positive cystoscopy results for recurrence or suspect areas underwent transurethral resection of the bladder. Cystoscopy was considered the reference standard to establish the bladder tumor diagnosis. The patients were divided into low, intermediate, and high risk groups for progression according to the European Association of Urology criteria. RESULTS: A total of 195 patients were suitable for evaluation. The urine samples from 21 patients were considered not evaluable for ImmunoCyt/uCyt+, and those patients were excluded from the study. Of all the control patients, 69.7% had negative cystoscopy findings. In the low-risk group, 84 patients underwent 131 cystoscopies, which diagnosed 30 Stage pTaG1 recurrences but no progression. Cytology and ImmunoCyt/uCyt+ together had a sensitivity of 86.6% and a negative predictive value of 95.2%. CONCLUSIONS: Of the cystoscopies performed during the 26 months of follow-up, 69.7% were negative. In the low-risk group, 30 Stage pTaG1 tumors and no progression was detected at a total cost of 14,672 Euros (USD 17,606). ImmunoCyt/uCyt+ and cytology every 6 months combined with annual cystoscopy reduced the morbidity and cost of follow-up in this group. PMID- 16698356 TI - Prospective study evaluating efficacy and safety of Adjustable Continence Therapy (ProACT) for post radical prostatectomy urinary incontinence. AB - OBJECTIVES: To examine a new prosthesis, the Adjustable Continence Therapy (ProACT), to determine its ability to treat effectively post radical prostatectomy urinary incontinence. Urinary incontinence is one of the most significant complications of radical prostatectomy. Although the artificial urinary sphincter (AUS) is considered the standard treatment for this condition, many men seek a simpler and less expensive treatment option. METHODS: From November 2000 to March 2004, 25 patients with severe post radical prostatectomy urinary incontinence were treated using the ProACT device. The preoperative evaluation included pad count, Valsalva leak point pressure determination, and Incontinence Quality-of-Life scores. In the follow-up, the same parameters, as well as complications, were analyzed and compared with the baseline measurements to assess the efficacy. RESULTS: The follow-up period was 6 to 48 months (mean 22.4). Of the 25 patients, 23 had follow-up data available for analysis. The improvements in pad count, Incontinence Quality-of-Life score, and Valsalva leak point pressures from baseline to the last follow-up examination were all significant (P <0.05). Overall, of the 23 patients followed up, 15 (65.2%) were continent using 0 to 1 pad daily and satisfied, 3 (13%) were improved but unsatisfied, and 5 (22%) did not have any improvement. Balloon adjustments were performed in all patents to achieve continence. Revision surgery was required in 4 (17%) of 23 patients. CONCLUSIONS: The use of ProACT represents a safe and effective treatment for post radical prostatectomy incontinence with a good degree of patient satisfaction and a low complication rate. Postoperative adjustments were necessary in most patients and were undertaken as a simple outpatient visit. PMID- 16698357 TI - Robotic repair of vesicovaginal fistula: case series of five patients. AB - OBJECTIVES: To describe a technique of robotic repair of vesicovaginal fistula (VVF) and present our experience with 5 such patients. METHODS: A total of 5 patients were diagnosed with posthysterectomy (n = 4) or postmyomectomy (n = 1) VVF. All patients were first treated conservatively with continuous drainage using a Foley catheter without any success. After 12 weeks, these patients underwent robotic repair of the VVF. The steps of the technique of robotic repair are (a) vaginoscopy, (b) cystoscopy, (c) bilateral ureteral catheterization, (d) placement of ports for robotic repair, (e) peritoneoscopy, (f) lysis of adhesions, (g) incision of the bladder and cystotomy in reverse tennis racquet fashion encircling the fistula, (h) excision and freshening of the fistulous margins after complete separation of the bladder from the vagina, (i) closure of the vaginal opening horizontally and bladder opening vertically with interrupted Vicryl sutures, and, finally, (j) interposition of the omentum between these suture lines. RESULTS: Fistula repair was successful in all cases, with a mean operative time (from cystoscopy to the end of the procedure) of 233 minutes (range 150 to 330) and estimated blood loss of less than 70 mL. The length of hospital stay was a mean of 5 days (range 4 to 7). The Foley catheter was removed on the 10th postoperative day after voiding cystourethrography. At 6 months of follow-up, these patients continued to void normally without any recurrence of VVF. CONCLUSIONS: These data suggest that robot-assisted VVF repair is feasible and results in lower morbidity, a shorter hospital stay, and a quicker recovery. The minimally invasive approach of robot-assisted VVF repair may be a more attractive option for patients with VVF. PMID- 16698358 TI - Significance of nocturnal hesitancy in treatment of men with lower urinary tract symptoms. AB - OBJECTIVES: To determine the significance of nocturnal hesitancy in the treatment of men with lower urinary tract symptoms. METHODS: A total of 123 patients with nocturia were prospectively studied. The nocturnal hesitancy scores ranged from 0 to 5 and were scored according to the International Prostate Symptom Score. The patients were stratified on the basis of their nocturnal hesitancy scores as group 1 (0 to 1, n = 57), group 2 (2 to 3, n = 45), and group 3 (4 to 5, n = 21). All patients received treatment with an alpha-blocker once daily for 4 weeks. RESULTS: Group 3 scored significantly higher on the International Prostate Symptom Score at baseline and after treatment than did group 1. The actual number of nightly voids was greatest in group 3 and was lowest in group 1 (P = 0.011 at baseline and P = 0.046 after treatment, respectively). The baseline nocturia indexes were greatest in group 3 and were lowest in group 2 (P = 0.027). A significant but weak correlation was also noted between the nocturia hesitancy score and the differences in scores between the posttreatment and baseline voiding symptoms (r = -0.234, P = 0.021) and total International Prostate Symptom Score (r = -0.270, P = 0.011), respectively. In our multivariate analyses, the straining score was a significant determinant of nocturnal hesitancy. CONCLUSIONS: The results of our study revealed that more emphasis should be placed on nocturnal hesitancy in the terminology of lower urinary tract symptoms. Additional research regarding the pathophysiologic mechanisms underlying nocturnal hesitancy, as well as its effects on those with it, is clearly warranted. PMID- 16698359 TI - Positron emission tomography with C11-acetate for tumor detection and localization in patients with prostate-specific antigen relapse after radical prostatectomy. AB - OBJECTIVES: To evaluate positron emission tomography with C11-acetate as a method for detecting and localizing prostate cancer recurrence. No technique for localizing and detecting prostate cancer recurrence after biochemical relapse available today is sensitive enough to localize recurrence at a stage at which salvage radiotherapy is still curative. METHODS: Twenty patients (age 56 to 77 years) who had undergone radical prostatectomy and had an increasing prostate specific antigen level measured on two consecutive occasions were included. In addition to the investigations usually performed when prostate cancer recurrence is suspected, they underwent positron emission tomography with C11-acetate as the marker. RESULTS: Pathologic uptake of acetate was seen in 15 (75%) of the 20 patients. In 8 of these patients, a solitary lesion was found (seven in the prostatic fossa and one at the regional lymph nodes). Multiple lesions were found in the remaining 7. False-positive uptake was seen in 3 men (15%). Additional investigations in these men revealed pathologic findings other than prostate cancer. CONCLUSIONS: Positron emission tomography with C11-acetate as marker is a promising method for early detection and localization of prostate cancer recurrence. False-positive uptake does occur. PMID- 16698360 TI - Phase II study evaluating oral triamcinolone in patients with androgen independent prostate cancer. AB - OBJECTIVES: To assess the effect of triamcinolone administration on the serum prostate-specific antigen (PSA) response and the time to progression in patients with androgen-independent prostate cancer (AIPC). METHODS: Patients with AIPC were prospectively treated with oral triamcinolone 4 mg twice daily, and their serum PSA and cortisol levels were measured monthly. Patients with greater than 25% increases in serum PSA from baseline were considered to have progressive disease and were removed from the study. Those patients who had a decrease in serum PSA levels or stable disease continued in the study until disease progression. Bone scans were obtained every 12 weeks and at progression. RESULTS: Twenty-four patients with AIPC were treated from November 2002 to June 2004. A partial response with a more than 50% decrease in serum PSA level was seen in 29%. Another 21% achieved stable disease. No statistically significant difference was found in the time to progression in the partial responders and patients with stable disease. The median time to progression in both groups was 7.5 months. Treatment was well tolerated without any grade 3 or 4 toxicity. CONCLUSIONS: Oral triamcinolone was well tolerated by patients with AIPC, with 50% of the patients exhibiting a good response to therapy in terms of serum PSA level and time to progression. PMID- 16698361 TI - Prognostic factors in lymph node-positive prostate cancer. AB - OBJECTIVES: To characterize lymph node metastasis of prostate cancer (PCa) and identify the parameters associated with patient outcome. The incidence of clinically localized PCa with concurrent lymph node metastasis has decreased to less than 1% in the United States but is between 10% and 15% in other countries. METHODS: Our study cohort of 1148 patients underwent radical prostatectomy in Ulm, Germany, between 1986 and 2002, and 201 (18%) had lymph node-positive PCa. RESULTS: The metastases showed growth architecture resembling primary PCa. We assigned a Gleason pattern and evaluated for size, extranodal extension, and lymphovascular invasion (LVI). Of 201 patients, 155 had original pathology slides available; 36 of the 155 were excluded because of preoperative hormonal ablation therapy. Of the remaining 119 patients, 22 (19%) were assigned Gleason pattern 3, 93 (78%) Gleason pattern 4, and 4 (3%) Gleason pattern 5. Extranodal extension was present in 66 (55%) of 119 patients and LVI in 29 (25%). An increased risk of prostate-specific antigen (PSA) recurrence was found for Gleason pattern 4/5 (hazard ratio [HR] 2.5, P = 0.038), LVI in the lymph nodes (HR 1.9, P = 0.038), and nuclear grade of the primary tumor (HR 2, P = 0.025). Independent predictors of PSA recurrence included LVI and nuclear grade (HR 1.9, P = 0.03 and HR 2, P = 0.03, respectively). CONCLUSIONS: Lymph node metastases of PCa are heterogeneous and have a close relation to the corresponding primary tumor. Most patients with lymph node-positive PCa remained disease free for up to 13 years after radical prostatectomy. Independent predictors of PSA recurrence among those with lymph node-positive PCa included LVI in the lymph nodes and the nuclear grade of the primary tumor. These parameters may be useful in predicting PSA recurrence in lymph node-positive PCa and could be included in patient follow-up. PMID- 16698362 TI - Ethnicity and determinants of quality of life after prostate cancer treatment. AB - OBJECTIVES: To examine the quality of life (QOL) in a sample of men recently treated for localized prostate cancer to determine whether minority men are at greater risk of decrements in QOL and to identify factors that might explain disparities in QOL outcomes. METHODS: The relationship between ethnicity and QOL was evaluated in a diverse sample of 204 men (85 non-Hispanic white, 37 African American, and 82 Hispanic men). We also assessed associations with other factors known to be related to QOL (ie, sociodemographic, medical, and health behavior factors). Hierarchical regression analysis was used to assess the relationship between ethnicity and QOL. Factors that were anticipated to explain the ethnic differences in QOL were then added in stepwise analyses. RESULTS: Ethnic group membership was related to QOL such that minority men had lower QOL than non Hispanic white men. In subsequent steps, the association between ethnic group membership and QOL was partially mediated by sociodemographic, medical, and health behavior factors, with each factor adding significant incremental variance (5%, 5%, and 17%, respectively). Three variables remained significant in the final model, which explained 37% of the variance in QOL scores: medical comorbidity, physical activity, and sleep functioning. CONCLUSIONS: Health behaviors appear to be strongly related to men's QOL after prostate cancer treatment. Intervention studies aimed at improving QOL should include a brief assessment of health behaviors and may want to incorporate intervention components designed to address physical activity and sleep functioning. PMID- 16698363 TI - Outcome after radical prostatectomy in young men with or without a family history of prostate cancer. AB - OBJECTIVES: To establish the cancer control afforded by radical prostatectomy in patients aged 50 years or younger with sporadic, familial, or hereditary prostate cancer. METHODS: We collected data on all patients 50 years old or younger among 5880 patients treated for prostate cancer between 1994 and 2004. We recorded age, ethnic origin, clinical presentation, family history of prostate cancer, preoperative prostate-specific antigen (PSA) level, treatment, Gleason score, 2002 TNM stage, surgical margin status, and disease progression. Radical prostatectomy was offered as first-line treatment to all patients with localized prostate cancer (T1-T2N0M0) and negative lymph nodes. PSA-free survival was calculated. RESULTS: We analyzed the data from 110 patients (1.9%), of whom 37 had familial cancer (33.6%) and 15 hereditary cancer (13.6%). A total of 85 patients (77.3%) had undergone radical prostatectomy, 39 (45.9%) by open retropubic surgery and 46 (54.1%) laparoscopically. The surgical margins were positive in 11 patients (12.9%). The mean follow-up after prostatectomy was 39.1 +/- 36.8 months (range 4 to 125). Nine patients (10.6%) experienced biochemical recurrence (PSA level greater than 0.2 ng/mL). Longer PSA-free survival after surgery was significantly associated with high-risk and intermediate-risk patients (P = 0.01 and P = 0.02, respectively) but not with the surgical procedure (P = 0.6) or family history of cancer (P = 0.46). CONCLUSIONS: Radical prostatectomy is an effective treatment of localized cancer in patients younger than 50 years old. Nearly one half of our cases of prostate cancer in younger men were forms of familial cancer. Detecting familial forms is a key objective in early screening and in the timely identification of candidates for prostatectomy. PMID- 16698364 TI - Defining association between sleep apnea syndrome and erectile dysfunction. AB - OBJECTIVES: To conduct a study using validated sexual function and sleepiness inventories to define whether sleep apnea syndrome (SAS) is associated with erectile dysfunction and whether any correlation exists between the severity of SAS and the severity of erectile dysfunction. Previous work has suggested that sleep disorders are associated with erectile dysfunction. METHODS: Men presenting to a sleep clinic with symptoms consistent with SAS were given the Epworth Sleepiness Scale and an erectile dysfunction risk factor inventory, the International Index of Erectile Function. A database was constructed and statistical analysis conducted to define the correlation between the two entities. RESULTS: A total of 50 men met the criteria for inclusion. Of the 50 men, 60% had abnormal Epworth Sleepiness Scale scores and 80% of these patients had erectile dysfunction as determined by inventory scores compared with 20% of the men with normal Epworth Sleepiness Scale scores. There were statistically significant differences between men with normal and abnormal sleepiness scores for the total and erectile function domain of the International Index of Erectile Function. The correlation between the severity of the sleepiness and the severity of erectile dysfunction was good (r = -0.80, P = 0.012). CONCLUSIONS: Men presenting with symptoms consistent with SAS have a significant risk of erectile dysfunction, and the correlation between the severity of sleep apnea and the severity of erectile dysfunction is strong. PMID- 16698365 TI - Long-term oral phosphodiesterase 5 inhibitor therapy alleviates recurrent priapism. AB - OBJECTIVES: Recurrent ischemic priapism describes a disorder of repeated episodes of prolonged penile erection that frequently leads to devastating complications of erectile tissue damage and erectile dysfunction. A mechanistic role for dysregulated phosphodiesterase 5 (PDE5) in the deranged smooth muscle response of the corpus cavernosum of the penis offers new understanding about the pathogenesis of the disorder and suggests that PDE5 may serve as a molecular target for its treatment and prevention. We explored the use of PDE5 inhibitors to treat recurrent priapism, based on the hypothesis that the erection regulatory function of PDE5 would be regularized by this treatment and protect against further episodes. METHODS: We administered PDE5 inhibitors using a long-term therapeutic regimen to 3 men with sickle cell disease-associated priapism recurrences and 1 man with idiopathic priapism recurrences. RESULTS: Long-term PDE5 inhibitor treatment alleviated priapism recurrences. CONCLUSIONS: These observations support the hypothesis that PDE5 dysregulation exerts a pathogenic role for priapism associated with hematologic dyscrasias, as well as idiopathic priapism. Although these preliminary findings suggest that continuous, long-term PDE5 inhibitor therapy may be useful as a preventative strategy for priapism, additional evaluation in the form of a controlled clinical trial is needed. PMID- 16698366 TI - Frequency of wetting is predictive of response to anticholinergic treatment in children with overactive bladder. AB - OBJECTIVES: To examine the variables relative to the response of oxybutynin treatment in children with daytime urinary incontinence. METHODS: The records of patients seen for voiding problems between 1999 and 2003 were reviewed. Patients taking oxybutynin for 3 months or longer were included in the study. Patients with structural or neurologic bladder abnormalities and those taking oxybutynin at the initial visit were excluded. Age, sex, uroflowmetry findings, postvoid residual urine volume, duration and severity of symptoms before oxybutynin, urinary tract infection history, constipation, and the duration, dosage, and adverse effects of oxybutynin treatment were evaluated. Data were analyzed using Fisher's exact test for categorical variables, the two-sample t test or Wilcoxon rank-sum test for continuous and ordinal variables, and the linear mixed model analysis for uroflow data. RESULTS: Eighty-one patients met the inclusion criteria. After an average follow-up of 1.2 years, 31 (38.3%) were dry, 25 (30.9%) had experienced significant improvement, 19 (23.5%) had experienced slight improvement, and 6 (7.4%) were unchanged. No significant differences were detected between those who became dry and those with little to no improvement with respect to age, sex, duration of symptoms, follow-up, uroflow pattern, postvoid residual urine volume, or bladder capacity. Those children presenting with decreased frequency of wetting episodes were significantly more likely to obtain daytime continence. The most common side effects were constipation (18.5%), dry mouth (17.3%), and flushing (13.6%). CONCLUSIONS: Children with daytime incontinence presenting with the lowest frequency of wetting were most likely to achieve continence. The frequency of wetting should be considered a significant prognostic variable when assessing the results of therapeutic intervention trials. PMID- 16698369 TI - Role of positional instillation of contrast cystography in the algorithm for evaluating children with confirmed pyelonephritis. AB - OBJECTIVES: To assess the utility of positional instillation of contrast (PIC) cystography in detecting vesicoureteral reflux (VUR) in patients with renal scarring from recurrent febrile urinary tract infections that standard voiding cystourethrography and nuclear cystogram imaging failed to reveal. METHODS: Between June 2004 and November 2004, a total of 5 pediatric patients with recurrent febrile urinary tract infections and radiologic evidence of upper tract involvement were examined with PIC cystography. All patients had at least one previous negative standard reflux study (voiding cystourethrography or nuclear cystography). RESULTS: All 5 patients showed VUR on PIC cystography. Unilateral reflux was detected in 3 patients, and 2 patients had bilateral VUR. CONCLUSIONS: The PIC cystogram should be integrated into the algorithm for diagnosing patients with recurrent febrile urinary tract infection, who do not exhibit VUR on standard imaging modalities. The morbidity associated with undiagnosed VUR, as demonstrated by this group of patients, suggests that permanent renal damage may be prevented by early diagnosis and treatment. PMID- 16698373 TI - Dysfunction of lower urinary tract in renal transplant children with nephrologic disease. AB - OBJECTIVES: To investigate the relationship between dysfunction of the lower urinary tract after renal transplantation and renal transplant function in children with an underlying nephrologic disease. METHODS: The research group consisted of 21 renal transplant children (12 girls and 9 boys, mean age 13.5 years, range 6 to 18) with an underlying nephrologic disease. To indicate renal transplant function, the calculated creatinine clearance rate (Ccr) according to Schwartz was used. The Ccr was measured at two points, 2 months after transplantation and at the moment of study. The average graft age was 34 months (range 5 to 85). The data on dysfunction of the lower urinary tract were gathered using a written questionnaire, frequency volume chart, free uroflowmetry, transabdominal ultrasonography, and medical records. To determine the relationship between the symptoms of dysfunction of the lower urinary tract and Ccr at the moment of study, we computed bivariate correlations and performed multivariate regression analyses in which the associations were studied while controlling for the Ccr 2 months after transplantation and graft age. RESULTS: A sensation of incomplete emptying (P = 0.03), postvoid residual urine volume (P = 0.06), and urinary tract infection (P = 0.004) correlated negatively with the Ccr at the moment of study. These effects remained present (P = 0.07, P = 0.03, and P = 0.003, respectively) while controlling for graft age and the Ccr at 2 months after transplantation in the regression analysis. CONCLUSIONS: The results of our study have shown that a postvoid residual urine volume and urinary tract infections after renal transplantation may result in renal transplant deterioration in children with an underlying nephrologic disease. PMID- 16698375 TI - Lift-assisted offset lithotomy: positioning for transurethral operations in morbidly obese. AB - The incidence of morbid obesity is rapidly increasing in the United States. This presents a unique challenge in the diagnosis, management, and surgical treatment of urologic disease. The use of the lithotomy position for transurethral procedures is often not possible because of the body habitus and equipment specific weight limitations. During positioning, it becomes imperative to maintain the safety of both the patient and the operating room personnel. We present a new, bariatric lift-assisted positioning technique used in the management of a 311-kg morbidly obese man with high-grade, superficial, papillary urothelial carcinoma. PMID- 16698376 TI - Renal calculi presenting as hyperhidrosis in patient with spinal cord injury. AB - Renal calculi in the spinal cord injury population present a diagnostic dilemma for urologists. However, 7% of all patients with spinal cord injury will develop renal calculi. Undiagnosed stone disease can lead to significant morbidity and mortality in this population. This case demonstrates the subtlety of the presenting symptoms of renal calculi in the patient with spinal cord injury. PMID- 16698378 TI - Giant accessory female phallic urethral diverticulum. AB - Anomalies of the urogenital sinus have been described in association with male pseudohermaphroditism, bladder and uterine duplication, and a spectrum of external genital anomalies, including a female phallic urethra. The evaluation and optimal surgical management of these anomalies can be complex. We describe a case of a female patient with an accessory phallic urethra and a urogenital sinus anomaly. The presence of stenosis of the accessory urethral meatus led to the formation of a giant diverticulum with unusual features. The complete repair of the anomaly incorporated creation of labia minora from portions of the diverticulum. PMID- 16698377 TI - Failure of gonadotropin-releasing hormone agonists with and without sterile abscess formation at depot sites: insight into mechanisms? AB - We describe a patient with advanced prostate cancer who failed to achieve testosterone suppression with depot leuprolide after developing sterile abscesses at the injection sites. When the patient was switched to depot goserelin, he did not have any evidence of inflammation at the injection sites, but testosterone suppression again failed. This case suggests variable mechanisms for failure of gonadotropin-releasing hormone agonist therapy and highlights the necessity of prospective testosterone monitoring in patients who have developed sterile abscesses, even if switched to another gonadotropin-releasing hormone agonist. PMID- 16698379 TI - Resection of recurrent inferior vena cava tumor after radical nephrectomy for renal cell carcinoma. AB - Management of recurrent tumor in the inferior vena cava (IVC) after radical nephrectomy is surgically challenging. We report 3 cases of recurrent renal cell carcinoma within the IVC managed by three different surgical techniques. One patient was treated with tumor thrombus removal and primary cavotomy closure. The second patient was treated with IVC ligation and removal without vascular reconstruction. A third patient was treated with IVC wall excision and placement of a bovine pericardium graft. Although technically difficult, repeat resection of IVC tumor recurrence after nephrectomy for renal cell carcinoma is an acceptable method of treatment. PMID- 16698380 TI - Xanthine urolithiasis. AB - Xanthine calculi are uncommonly encountered stones. When they occur, they typically do so in association with inborn metabolic disorders such as hereditary xanthinuria or Lesch-Nyhan syndrome. They may also occur in association with states of profound hyperuricemia such as myeloproliferative disease after treatment with allopurinol. If the underlying disorder is not addressed, a high risk of stone recurrence exists. Therefore, to raise clinical awareness, we reviewed and report our experience in the treatment of patients with these stones, discussing the underlying pathophysiology and approach to treatment. PMID- 16698381 TI - Fibrin sealant for retrograde ureteroscopic closure of urine leak after partial nephrectomy. AB - We present a case of a prolonged urine leak after partial nephrectomy successfully treated by retrograde ureteroscopic injection of fibrin sealant. The technical aspects of retrograde instillation of sealant into the lower renal pole and outcome are discussed. PMID- 16698382 TI - Refractory hematuria from amyloidosis successfully treated by splenectomy. AB - Systemic amyloidosis can result in a coagulopathy that is associated with low levels of factor X. We present a case of intractable, life-threatening hematuria that was successfully managed with activated recombinant human factor VII and splenectomy. PMID- 16698383 TI - Presentation of fused vas deferens. AB - Transverse testicular ectopia is a rare condition that usually presents as a unilateral nonpalpable testis and a contralateral descended testis with an associated hernia. Currently, with the use of diagnostic laparoscopy, transverse testicular ectopia is found before groin exploration. We reviewed 2 cases that were referred to our institution. Both patients had a common vas deferens with proximal fusion. To our knowledge, a fused vas deferens has only been reported three other times in published reports. Our second case is unique in that this is the first time a common vas deferens has been reported outside of transverse testicular ectopia. PMID- 16698384 TI - Ovarian hyperstimulation causing ureteral obstruction in a pregnant woman. AB - Obstruction of the urinary collecting system in pregnant women can present a unique challenge to the urologist. Although ureteral obstruction due to stone disease is well-documented and ureteral obstruction is a well-known and defined cause of acute renal failure, obstruction from extrinsic compression is a less understood phenomenon. We report a case of ureteral obstruction from hyperplastic ovaries in a pregnant woman receiving hormonal therapy that was managed initially with ureteral stenting. PMID- 16698385 TI - Retroperitoneal ganglioneuroma presenting as right renal mass. AB - A primary retroperitoneal ganglioneuroma was found in a 37-year-old woman on computed tomography, which revealed a homogeneous mass just above the right kidney. The tumor was resected through a transabdominal approach. The resected specimen measured 11 x 9 x 4.5 cm and weighed 270 g. The histologic examination showed that the lesion was a ganglioneuroma composed of both mature ganglion cells and nerve fibers. PMID- 16698386 TI - Urethral polyp in asymptomatic male infant with prenatal hydronephrosis. AB - Congenital posterior urethral polyps are rare, benign lesions of the posterior urethra. We report a 1-day-old newborn boy with a solitary fibroepithelial posterior urethral polyp who presented with a history of prenatal mild hydronephrosis, subsequently associated with bladder wall thickening on ultrasonography. PMID- 16698387 TI - Association of interleukin-1Ra gene polymorphism in patients with bladder cancer: case control study from North India. AB - OBJECTIVES: To evaluate whether polymorphism of interleukin (IL)-1beta gene (exon 5 and promoter region) and IL-1 receptor antagonist (Ra), 86-bp variable number tandem repeat, are associated with transitional cell carcinoma of the urinary bladder, because cytokines have been hypothesized to be important in cancer development. METHODS: The study included 120 patients with bladder cancer (age range 32 to 69 years) and 150 age-matched controls (age range 25 to 62 years). The polymorphisms were identified by polymerase chain reaction/restriction fragment length polymorphism method and IL-1Ra polymorphism by variable number of identical tandem repeat analysis. Genotype distribution and allelic frequencies between patients and controls were compared. RESULTS: A significant difference was found in the frequency distribution of the IL-1Ra gene polymorphism in patients with bladder cancer compared with the normal control group (P < 0.001), but no difference was found in the frequencies of the IL-1beta promoter region and exon 5 genotypes between patients with bladder cancer and controls (P = 0.112 and P = 0.953, respectively). CONCLUSIONS: This is perhaps the first report on polymorphic changes in gene encoding IL-1Ra in patients with bladder cancer from India. Our data suggest that IL-1Ra intron 2 polymorphism seems to play a prominent role among the IL-1 gene cluster with respect to bladder cancer, and the association studies appear to be plausible in determining the cancer susceptibility and risk. PMID- 16698388 TI - Time-dependent variations in inflammation and scar formation of six different pubovaginal sling materials in the rabbit model. AB - OBJECTIVES: To provide pathologic evidence, using six different sling materials, of the findings from rabbit model studies demonstrating loss of tensile strength and stiffness in porcine and cadaveric sling materials. METHODS: Ten rabbits randomized into two survival groups (6 and 12 weeks of age) each had human cadaveric fascia, porcine dermis, porcine small intestine submucosa, polypropylene mesh, and autologous fascia implanted on their anterior rectus fascia. At harvest, hematoxylin-eosin and immunohistochemical staining for CD3, CD20, and MIB-I were performed. A pathologist unaware of the content of the slides quantified the degree of inflammation and fibrosis of each. RESULTS: Significant differences were found for inflammation (P = 0.016), eosinophil infiltrate (P = 0.035), and inflammatory rind (P = 0.027) at 12 weeks, with polypropylene mesh having the lowest degree. At 12 weeks, differences were found in the presence of fibrosis/scar formation (P = 0.010) and degree of fibrosis/scar (P = 0.009). Although polypropylene mesh, cadaveric fascia, and porcine dermis all demonstrated a high presence of fibrosis/scar, polypropylene mesh had the greatest overall degree of scar formation at 12 weeks. CONCLUSIONS: The inflammation with the cadaveric fascia and porcine materials may cause rapid clinical deterioration compared with autologous fascia and polypropylene mesh. These data provide a possible explanation for prior biomechanical studies demonstrating variations in tensile strength and stiffness of the different materials. The fibrosis and scarring noted with polypropylene mesh may also contribute to a more lasting repair. PMID- 16698389 TI - Curbing the obesity epidemic. PMID- 16698390 TI - Condoms and the Vatican. PMID- 16698391 TI - A reappraisal of H5N1 avian influenza. PMID- 16698392 TI - Deeper lessons from the CMAJ debacle. PMID- 16698393 TI - Dip-stick test for trachoma control programmes. PMID- 16698394 TI - Emergency management of seizures in children. PMID- 16698395 TI - Worms, asthma, and the hygiene hypothesis. PMID- 16698396 TI - Surgery for ovarian cancer: how to improve survival. PMID- 16698397 TI - Global framework on essential health R&D. PMID- 16698398 TI - Jules Rimet's bananas. PMID- 16698399 TI - Frank Artress. PMID- 16698400 TI - Cuttings. PMID- 16698401 TI - Antivirals for influenza in healthy adults. PMID- 16698402 TI - Antivirals for influenza in healthy adults. PMID- 16698403 TI - Antivirals for influenza in healthy adults. PMID- 16698404 TI - Chronic fatigue syndrome. PMID- 16698406 TI - Chronic fatigue syndrome. PMID- 16698407 TI - Vaccination in Australia. PMID- 16698409 TI - Drug-resistant enteric fever in the UK. PMID- 16698410 TI - Aminophylline in bradyasystolic cardiac arrest: a randomised placebo-controlled trial. AB - BACKGROUND: Endogenous adenosine might cause or perpetuate bradyasystole. Our aim was to determine whether aminophylline, an adenosine antagonist, increases the rate of return of spontaneous circulation (ROSC) after out-of-hospital cardiac arrest. METHODS: In a double-blind trial, we randomly assigned 971 patients older than 16 years with asystole or pulseless electrical activity at fewer than 60 beats per minute, and who were unresponsive to initial treatment with epinephrine and atropine, to receive intravenous aminophylline (250 mg, and an additional 250 mg if necessary) (n=486) or placebo (n=485). The patients were enrolled between January, 2001 and September, 2003, from 1886 people who had had cardiac arrests. Standard resuscitation measures were used for at least 10 mins after the study drug was administered. Analysis was by intention-to-treat. This trial is registered with the ClinicalTrials.gov registry with the number NCT00312273. FINDINGS: Baseline characteristics and survival predictors were similar in both groups. The median time from the arrival of the advanced life-support paramedic team to study drug administration was 13 min. The proportion of patients who had an ROSC was 24.5% in the aminophylline group and 23.7% in the placebo group (difference 0.8%; 95% CI -4.6% to 6.2%; p=0.778). The proportion of patients with non-sinus tachyarrhythmias after study drug administration was 34.6% in the aminophylline group and 26.2% in the placebo group (p=0.004). Survival to hospital admission and survival to hospital discharge were not significantly different between the groups. A multivariate logistic regression analysis showed no evidence of a significant subgroup or interactive effect from aminophylline. INTERPRETATION: Although aminophylline increases non-sinus tachyarrhythmias, we noted no evidence that it significantly increases the proportion of patients who achieve ROSC after bradyasystolic cardiac arrest. PMID- 16698411 TI - Field evaluation of a rapid point-of-care assay for targeting antibiotic treatment for trachoma control: a comparative study. AB - BACKGROUND: Trachoma results from repeated episodes of conjunctival infection with Chlamydia trachomatis and is the leading infectious cause of blindness. To eliminate trachoma, control programmes use the SAFE strategy (Surgery, Antibiotics, Face cleanliness, and Environmental improvement). The A component is designed to treat C trachomatis infection, and is initiated on the basis of the prevalence of the clinical sign trachomatous inflammation-follicular (TF). Unfortunately, TF correlates poorly with C trachomatis infection. We sought to assess a newly developed point-of-care (POC) assay compared with presence of TF for guiding the use of antibiotics for trachoma control. METHODS: We compared performance outcomes of the POC assay and presence of TF using commercial PCR as a comparator in 664 children aged 1-9 years in remote, trachoma-endemic villages in Tanzania. Signs of trachoma were graded according to the WHO simplified trachoma grading system. FINDINGS: Of 664 participants, 128 (19%) were positive for ocular C trachomatis infection by PCR. Presence of TF had a sensitivity of 64.1% (95% CI 55.8-72.4), specificity of 80.2% (76.8-83.6), and positive predictive value of 43.6% (36.5-50.7). By contrast, the POC assay had a sensitivity of 83.6% (77.2-90.0), specificity of 99.4% (98.8-100.0), and positive predictive value of 97.3% (94.2-100.3). Interagreements and intra-agreements between four novice operators were 0.988 (0.973-1.000) and 0.950 (0.894-1.000), respectively. INTERPRETATION: The POC assay is substantially more accurate than TF prevalence in identifying the presence or absence of infection. Additional studies should assess the use of the assay in the planning and monitoring of trachoma control activities. PMID- 16698412 TI - Efficacy and safety of intranasal lorazepam versus intramuscular paraldehyde for protracted convulsions in children: an open randomised trial. AB - BACKGROUND: In sub-Saharan Africa, rectal diazepam or intramuscular paraldehyde are commonly used as first-line anticonvulsant agents in the emergency treatment of seizures in children. These treatments can be expensive and sometimes toxic. We aimed to assess a drug and delivery system that is potentially more effective, safer, and easier to administer than those presently in use. METHODS: We did an open randomised trial in a paediatric emergency department of a tertiary hospital in Malawi. 160 children aged over 2 months with seizures persisting for more than 5 min were randomly assigned to receive either intranasal lorazepam (100 microg/kg, n=80) or intramuscular paraldehyde (0.2 mL/kg, n=80). The primary outcome measure was whether the presenting seizure stopped with one dose of assigned anticonvulsant agent within 10 min of administration. The primary analysis was by intention-to-treat. This study is registered with ClinicalTrials.gov, number NCT00116064. FINDINGS: Intranasal lorazepam stopped convulsions within 10 min in 60 (75%) episodes treated (absolute risk 0.75, 95% CI 0.64-0.84), and intramuscular paraldehyde in 49 (61.3%; absolute risk 0.61, 95% CI 0.49-0.72). No clinically important cardiorespiratory events were seen in either group (95% binomial exact CI 0-4.5%), and all children finished the trial. INTERPRETATION: Intranasal lorazepam is effective, safe, and provides a less invasive alternative to intramuscular paraldehyde in children with protracted convulsions. The ease of use of this drug makes it an attractive and preferable prehospital treatment option. PMID- 16698413 TI - Effect of albendazole treatments on the prevalence of atopy in children living in communities endemic for geohelminth parasites: a cluster-randomised trial. AB - BACKGROUND: Epidemiological studies have shown inverse associations between geohelminth (intestinal helminth) infection and atopy, leading to the suggestion that geohelminths might protect against allergy. Periodic deworming of school children with anthelmintics is a widely implemented intervention and has raised concerns that such programmes could increase allergy. We investigated the effect of repeated anthelmintic treatments with albendazole over 12 months on the prevalence of atopy and clinical indices of allergy. METHODS: We did a cluster randomised controlled trial in schoolchildren from 68 rural schools. Children were randomly assigned by school to either albendazole (34 schools, 1164 children) every 2 months for 12 months, or to no intervention (34 schools, 1209 children). The intervention schools received a total of seven albendazole treatments. The primary outcome was atopy at 12 months (allergen skin-test reactivity), and analysis was by intention-to-treat for whole-school analyses and per protocol for children. This study is registered as an International Standard Randomised Controlled Trial, number ISRCTN61195515. FINDINGS: Data for analysis were available for all schools and from 67.4% (784 of 1164) and 70.1% (848 of 1209) of children in albendazole and no-treatment groups, respectively. Albendazole treatment caused large reductions in geohelminth prevalence over the study period (adjusted odds ratio 0.13, 95% CI 0.09-0.19, p<0.001), but there was no evidence that treatment was associated with an increase in atopy prevalence (0.97, 0.68-1.39, p=0.862), or clinical allergy (wheeze, 1.07, 0.54-2.11, p=0.848) in the albendazole compared with the no-treatment group. INTERPRETATION: We saw no increase in the prevalence of atopy or clinical allergy associated with albendazole treatment. Deworming programmes for schoolchildren are unlikely to be accompanied by an increase in allergy. PMID- 16698414 TI - Annular erythema of Sjogren's syndrome. PMID- 16698415 TI - Cushing's syndrome. AB - Cushing's syndrome results from lengthy and inappropriate exposure to excessive glucocorticoids. Untreated, it has significant morbidity and mortality. The syndrome remains a challenge to diagnose and manage. Here, we review the current understanding of pathogenesis, clinical features, diagnostic, and differential diagnostic approaches. We provide diagnostic algorithms and recommendations for management. PMID- 16698416 TI - Persistent postsurgical pain: risk factors and prevention. AB - Acute postoperative pain is followed by persistent pain in 10-50% of individuals after common operations, such as groin hernia repair, breast and thoracic surgery, leg amputation, and coronary artery bypass surgery. Since chronic pain can be severe in about 2-10% of these patients, persistent postsurgical pain represents a major, largely unrecognised clinical problem. Iatrogenic neuropathic pain is probably the most important cause of long-term postsurgical pain. Consequently, surgical techniques that avoid nerve damage should be applied whenever possible. Also, the effect of aggressive, early therapy for postoperative pain should be investigated, since the intensity of acute postoperative pain correlates with the risk of developing a persistent pain state. Finally, the role of genetic factors should be studied, since only a proportion of patients with intraoperative nerve damage develop chronic pain. Based on information about the molecular mechanisms that affect changes to the peripheral and central nervous system in neuropathic pain, several opportunities exist for multimodal pharmacological intervention. Here, we outline strategies for identification of patients at risk and for prevention and possible treatment of this important entity of chronic pain. PMID- 16698417 TI - Our patients remain with us: that is what experience means. PMID- 16698418 TI - Necrotising pneumonia complicating a nasal furuncle. PMID- 16698419 TI - Foxp3 expressing CD4+ CD25+ and CD8+CD28- T regulatory cells in the peripheral blood of patients with lung cancer and pleural mesothelioma. AB - The role of T regulatory (Treg) cells in human cancer has not yet been clarified. We assessed the presence and function of CD4+ and CD8+ Treg cell subsets in the peripheral blood of patients with lung cancer (LC) and pleural mesothelioma (PM). We found a low but significant increase in the number of CD4+ T cells with phenotype and functional features of Treg cells in LC patients compared to normal healthy controls (NHC). Furthermore, total CD4+ T cells from LC patients proliferated less than cells from controls, suggesting that the increase in the CD4+ Treg cell pool has functional importance. LC patients also showed an expansion of the CD8+CD28- T cell subset and these cells expressed Foxp3 mRNA, as recently observed in alloantigen-specific CD8+CD28- T suppressor cells. No variation of peripheral Treg cell subsets was found in patients with PM, a disease with a predominantly localized nature. However, the lack of correlation between cancer stage and the number or the function of peripheral Treg cells in LC patients refuted the hypothesis that these cells are involved in tumor spreading. A possible involvement of the peripheral Treg cell pool in cancer development and/or in inducing systemic immunosuppression in LC patients can be hypothesized. PMID- 16698420 TI - Genetic variation in IL-8 associated with increased risk and poor prognosis of breast carcinoma. AB - Interleukin-8 (IL-8), a potent chemoattractant, has been demonstrated to contribute to human cancer progression through its potential functions as a mitogenic, angiogenic, and motogenic factor. We designed a broad study to investigate whether genetic variation in IL-8 has implications for susceptibility to and prognosis in breast carcinoma. We used the allele-specific polymerase chain reaction to characterize the variation of the IL-8 promoter region for 308 unrelated Tunisian patients with breast carcinoma and 236 healthy control subjects. Associations of the clinicopathologic parameters and the genetic marker with the rates of the breast carcinoma-specific overall survival and the disease free survival were assessed using univariate and multivariate analyses. A significantly increased risk of breast carcinoma was associated with heterozygous IL-8 (-251) TA (OR=1.58, p=0.02) and homozygous IL-8 (-251) AA (OR=1.76, p=0.01) variants. A significant association between the IL-8 (-251) AA homozygous genotype and the aggressive phenotype of breast carcinoma as defined by the high histological grade, auxiliary's lymph node metastasis, and large tumor size was found. The IL-8 (-251) A allele manifested a significant association with decreased overall survival and disease-free survival for breast carcinoma patients. The polymorphism in the promoter region of the IL-8 gene may not only represent a marker for the increased risk of breast carcinoma but also predict the clinical outcome. PMID- 16698421 TI - Expression of Fas, FasL, and soluble Fas mRNA in endomyocardial biopsies of human cardiac allografts. AB - Apoptosis mediated by the Fas/Fas ligand (FasL) has been implicated in rejection of solid organ allografts and it has been recently proposed that soluble forms of Fas could interfere with this interaction, blocking apoptosis. The purpose of this study was to analyze intragraft Fas, FasL, and soluble Fas mRNA levels in relation to acute rejection in cardiac allografts in humans. mRNA levels were determined by quantitative reverse transcriptase-polymerase chain reaction in 42 samples of endomyocardial biopsies obtained from 18 cardiac transplant recipients within the first 6 months after transplantation. FasL and Fas mRNA levels were higher in biopsies with rejection than in biopsies without rejection, and no difference was observed in soluble Fas mRNA. During rejection, there was a positive correlation between the mRNA levels of Fas-FasL, Fas-soluble Fas, and FasL-soluble Fas. During quiescent periods, however, the only correlation observed was between Fas and soluble Fas mRNA levels. In conclusion, our findings do not suggest a role for soluble Fas, confirm the heightened expression of FasL, and indicate, for the first time, an increased expression of Fas in acute rejection of cardiac allografts. PMID- 16698422 TI - Serum levels of interleukin-6 and its soluble receptors in patients with hepatitis C virus infection. AB - Interleukin-6 (IL-6) is an important cytokine in liver regeneration, and elevated levels of IL-6 have been demonstrated in patients with chronic liver diseases (CLD). Many biological effects of IL-6 depend on naturally occurring soluble IL-6 receptors. In the present study we measured the concentrations of IL-6 and its soluble receptors in the sera of patients with CLD related to hepatitis C virus (HCV) infection. We studied 77 patients with varying degrees of HCV-related CLD. Serum levels of IL-6 and its soluble receptors (sIL-6R, sgp130) were measured by enzyme-linked immunosorbent assay. Serum IL-6 and sIL-6R were elevated in patients with CLD compared with healthy subjects. Serum levels of sgp130 did not differ between patients with chronic hepatitis and healthy subjects. However, in patients with liver cirrhosis, sgp130 was significantly elevated and was positively correlated with total bilirubin and negatively correlated with cholinesterase and prothrombin time. Our study demonstrated that in patients with HCV-related CLD, serum IL-6 and its soluble receptor levels are correlated with both liver function impairment and the degree of liver fibrosis. These observations suggest that the balance of IL-6 and its soluble receptors may correspond to the state of liver damage in patients with CLD. PMID- 16698424 TI - Transcriptional control of activation-induced cytidine deaminase and error-prone DNA polymerases is functionally mature in the B cells of infants at birth. AB - Somatic hypermutation (SHM) of immunoglobulin genes requires activation-induced cytidine deaminase (AID). The error-prone DNA polymerases, such as Pol eta, Pol zeta, and Pol iota, also have been implicated in the process. Human adult antibodies directed to microbial pathogens are increased in affinity and function compared with those of infants. Adult antibodies achieve this increased affinity through somatic mutations, which are lacking in the B cells of infants. It is unknown if infant B cells are capable of upregulating the cell machinery needed to introduce mutations after stimulation through the antigen receptor. We show here that infant B cells exhibit similar kinetics and magnitude of transcription of AID and pol eta genes and only marginally lower levels of pol iota and pol zeta genes after stimulation through the B cell receptor. These data suggest that the ability to upregulate gene transcription of enzymes mediating SHM is not a limiting determinant of the functional quality of infant antibody responses. PMID- 16698423 TI - Rotavirus-specific CD5+ B cells in young children exhibit a distinct antibody repertoire compared with CD5- B cells. AB - Antiviral antibody responses in infants are limited in quality. One reason for this finding could be that the majority of B cells in infants are CD5+ cells, a subset of B cells that is thought to contain cells expressing polyreactive, low affinity B cell receptors. We analyzed the rotavirus (RV)-specific antibody heavy chain variable region (VH) repertoire in CD5+ and CD5- B cells of four RV infected children between 10 and 19 months of age. We found that the RV-specific B cell repertoire in CD5+ cells was VH3 family biased, in contrast to the VH1/VH4 dominance seen in CD5- B cells. The immunodominant RV-specific gene segment in CD5- B cells was VH1-46, which is the dominant segment used in RV-specific peripheral blood B cells from infants and adults. In contrast, the immunodominant gene segment was VH3-23 in RV-specific CD5+ B cells, which is the dominant gene segment in randomly selected B cells. Both RV-specific CD5+ and RV-specific CD5- B cells from all children studied demonstrated very low frequencies of somatic mutations. In conclusion, CD5+ B cells in infants responding to RV use an antibody gene repertoire that differs from the virus-specific repertoire of CD5- B cells, and both CD5+ and CD5- RV-specific B cells exhibit a low frequency of somatic mutations. PMID- 16698425 TI - HLA-DPB1*0202 is associated with a predictor of good prognosis of Graves' disease in the Japanese. AB - Whereas most patients with Graves' disease (GD) have antibodies against the thyrotropin receptor, which are measured as thyrotropin-binding inhibitory immunoglobulin (TBII), the TBII of 10% or less of Japanese patients with GD is undetectable at the first visit and throughout the entire clinical course, and these patients tend to respond well to medications and follow the better clinical course. Therefore, the absence of TBII at the first visit may be a predictor of good prognosis. Ninety-seven patients with GD who had remained TBII negative for at least 2 years from the onset, as well as 142 typical TBII-positive GD patients, were examined to reveal the HLA-linked immunogenetic background for this predictor. Compared with a healthy control population, the frequencies of HLA-A*0206 (OR=2.17, p=9.73x10(-4)) and DPB1*0501 (OR=3.26, p=3.31x10(-7)) carriers were increased in the typical patients, whereas those of HLA-A*0201 (OR=2.16, p=1.92x10(-3)), A*0207 (OR=3.19, p=7.17x10(-4)), and DPB1*0202 (OR=3.13, p=3.97x10(-4)) were increased in the TBII-negative group. These two patient groups were associated with similar HLA-A alleles and different HLA-DPB1 alleles, suggesting the presence of two genetic factors for GD within the HLA region; one is HLA-A linked and may be related to thyroid organ specificity, the other is HLA-DP linked and may control the severity of autoimmunity. PMID- 16698426 TI - Polymorphism in the 5' upstream regulatory and 3' untranslated regions of the HLA G gene in relation to soluble HLA-G and IL-10 expression. AB - The nonclassical human leukocyte antigen (HLA) class Ib gene HLA-G may be important for the induction and maintenance of immune tolerance between the mother and the semi-allogeneic fetus during pregnancy. Expression of HLA-G can influence cytokine and cytotoxic T-lymphocyte responses. Different HLA-G mRNA isoform expression patterns have been associated with HLA-G polymorphism, especially with a 14-bp insertion deletion polymorphism in the 3' untranslated region (3'UTR) of the HLA-G gene. A significantly high level of interleukin-10 (IL-10) secretion is observed in homozygous +14/+14-bp HLA-G peripheral blood mononuclear cells after lipopolysaccharide (LPS) stimulation. This study finds that polymorphism in the 5' upstream regulatory region (5'URR) of the HLA-G gene may also be implicated in differences in IL-10 secretion. However, this may also be due to linkage disequilibrium with the 14-bp polymorphism. A single-nucleotide polymorphism located -477 bp from the start site of exon 1 had a significant association with IL-10 concentrations but not after correction (p=0.011; pc=0.154). This polymorphism is located next to a heat shock element. Eighteen 5' URR/3'-UTR HLA-G haplotypes were defined; one common homozygous genotype based on these haplotypes was significantly associated with a high IL-10 level after LPS stimulation compared to certain other genotypes. This study indicates that polymorphism in the 5'-URR of the HLA-G gene may have functional significance, although a new line of investigations is needed to elucidate these findings. PMID- 16698427 TI - Identification of novel chimpanzee MHC class I and II alleles using an improved sequence-based typing strategy. AB - The chimpanzee (Pan troglodytes) is a valuable model for the study of infection with hepatitis C virus and hepatitis B virus, to which only humans and chimpanzees are susceptible. Because the cellular immune response plays a crucial role in host defense against these viruses, the analysis of major histocompatibility complex (MHC) (Patr) class I and II allele diversity in chimpanzees is essential for immune response analysis and vaccine development. In the present study, we report a novel, rapid, and sensitive sequence-based typing strategy to identify polymorphisms of the principal Patr class I (Patr-A and Patr B) and Patr class II (Patr-DQB1 and Patr-DRB1) alleles. Using this method we identified seven novel Patr alleles in 17 chimpanzees, one of them present in 3 chimpanzees. Furthermore, we detected heterozygosity more rapidly and with higher sensitivity than was done with previous techniques that were based on reverse transcription and amplification of messenger RNA followed by molecular cloning and sequencing. PMID- 16698428 TI - The haplotype structure of the human major histocompatibility complex. AB - There is great interest in the use of single-nucleotide polymorphisms (SNPs) and linkage disequilibrium (LD) analysis to localize human disease genes. The results suggest that the human genome, including the major histocompatibility complex (MHC), consists largely of 5- to 200-kb blocks of sequence fixity between which random recombination occurs. Direct determination of MHC haplotypes from family studies also demonstrates similar-sized blocks, but otherwise gives a very different picture, with a third to a half of Caucasian haplotypes fixed from HLA B to HLA-DR/DQ (at least 1 Mb) as conserved extended haplotypes (CEHs), some of which encompass more than 3 Mb. These fixed haplotypes differ in frequency both in different Caucasian subpopulations and in Caucasian patients with HLA associated diseases, complicating disease susceptibility gene localization. The inherent inability of LD analysis to "see" DNA fixity beyond three markers contributes to the failure of SNP/LD analysis to define in detail or even detect CEHs in the MHC and probably elsewhere in the genome. More importantly, the use of statistical analysis, rather than direct haplotype determination and counting, fails to reveal the details of haplotype structure essential for gene localization. Given the oversimplified picture of the MHC (and probably the rest of the genome) provided only by SNP/LD-defined blocks, it is questionable whether this approach will be of great help in disease susceptibility gene localization or identification. PMID- 16698429 TI - KIR gene in ethnic and Mestizo populations from Mexico. AB - Killer cell immunoglobulin-like receptors are characterized by their great diversity of genes and alleles. Population studies have identified the presence of a broad variety of genotypes. In Mexico, there are diverse ethnic groups representing 9% of the total population and the rest is composed of Mestizos with a more varied biology. For the purpose of this study, genotyping was performed in Mestizos, in Mexico City inhabitants, and in three ethnic groups. The frequencies of genes KIR2DL2, 2DL5, 2DS1-3, 2DS5, and 3DS1 showed a greater variability in the groups studied. A total of 12 different genotypes were identified, the higher number for the Mestizos and the lower number for the Tarahumaras. Genotype 1 was found at a greater frequency in all the groups, except for the Tarahumaras, in which genotype 4 was more frequent. The frequency of genotypes 4 and 8 in Mexicans was higher than that for other populations analyzed. By subtyping of KIR3DL1, 3DL2, 2DL1, and 2DL3, two B haplotypes were identified in families; both were absent in Caucasian families. Our results indicated a greater diversity of genes in the Mestizos group than in the ethnic groups. PMID- 16698430 TI - MICA is associated with type 1 diabetes in the Belgian population, independent of HLA-DQ. AB - To ascertain association of MICA with type 1 diabetes (T1D) in the Belgian population, well-characterized antibody-positive patients were analyzed for MICA transmembrane gene polymorphism in both an association study and a nuclear family study. The frequency of MICA5 was significantly increased in the T1D patient group (18%) compared with the control population (12%, OR=1.6, pc<10(-3)), whereas MICA9 was decreased (11% versus 16%, OR=0.7, pc<0.01). A p value<10(-3) for the association of MICA conditional on HLA class II and p=0.01 for the conditional extended transmission disequilibrium test were obtained, indicating that MICA is associated with type 1 diabetes, independent of HLA-DQ. Analysis of estimated extended HLA-DQ-MICA haplotypes revealed individual effects of MICA alleles. The most significant effect was seen for MICA5 on the HLA-DQA1*03 DQB1*0302-MICA haplotype (OR=2.5, p<10(-3)). A significant protective effect was seen for the combination of DQA1*01-DQB1*0602/3 and MICA5.1 (OR=0.3, p<10(-3)). However, patients stratified according to the presence or absence of the different MICA alleles did not differ in terms of age at onset, sex, or other diabetes-related clinical and epidemiological data. In conclusion, MICA is associated with type 1 diabetes in the Belgian population and the observed association does not result from the HLA-DQ associated risk. PMID- 16698431 TI - DC-SIGN interacts with Mycobacterium leprae but sequence variation in this lectin is not associated with leprosy in the Pakistani population. AB - The C-type lectin DC-SIGN is involved in early interactions between human innate immune cells and a variety of pathogens. Here we sought to evaluate whether DC SIGN interacts with the leprosy bacillus, Mycobacterium leprae, and whether DC SIGN genetic variation influences the susceptibility and/or pathogenesis of the disease. A case-control study conducted in a cohort of 272 individuals revealed no association between DC-SIGN variation and leprosy. However, our results clearly show that DC-SIGN recognizes M. leprae, indicating that mycobacteria recognition by this lectin is not as narrowly restricted to the Mycobacterium tuberculosis complex as previously thought. Altogether, our results provide further elucidation of M. leprae interactions with the host innate immune cells and emphasize the importance of DC-SIGN in the early interactions between the human host and the infectious agents. PMID- 16698432 TI - A family based linkage analysis of HLA and 5-HTTLPR gene polymorphisms in Sardinian children with autism spectrum disorder. AB - Autism spectrum disorders (ASD) are characterized by a broad range in clinical presentation. Although a definite genetic cause has not yet been fully demonstrated, family based studies suggest that a multigenic pattern may be responsible for susceptibility, but most results are conflicting and have yet to be replicated. The purpose of this investigation was to analyze the linkage of the human leukocyte antigen (HLA) and the human serotonin transporter coding (5 HTTLPR) genes with ASD in a group of 37 families of Sardinian ethnicity in insular Italy. In 50% of these families, ASD is linked to HLA, and in the other 50% it is linked to 5-HTTLPR polymorphic genes; in other words, linkage to one or the other was evident in all cases. Despite a very homogenous genetic pattern being generally reported for Sardinians, the linkage observed with HLA and 5 HTTLPR genetic regions indicated a statistically defined heterogeneity (p=0.002). No allelic HLA or 5-HTTLPR polymorphisms were specifically associated with ASD, suggesting these loci as markers of other genes mapped in their close proximity that may be more directly involved and thus may merit further analytical studies. PMID- 16698433 TI - Killer cell immunoglobulin-like receptor (KIR) genes in the Basque population: association study of KIR gene contents with type 1 diabetes mellitus. AB - Killer cell immunoglobulin-like receptors (KIR) form a group of regulatory molecules that specifically recognize HLA class I molecules, modulating cytolytic activity of natural killer cells. The number of KIR genes can vary between individuals, significant allelic variations have been described, and KIR genes are organized in a complex and heterogeneous family. In the present study we have performed KIR genotype analysis in the Basque general population. Additionally, we have tested the possible association between KIR gene content (in combination with its HLA ligand) and type 1 diabetes mellitus (T1DM). KIR genotyping was performed using a commercial sequence-specific primer amplification genotyping kit and amino acid position 80 of HLA-C was genotyped by specific amplification and direct sequencing. Haplotypes and genotypes were deduced based on previous studies, and frequencies were compared between disease and control groups. All KIR genes tested were present in Basques and several of these genes (KIR2DS5, KIR3DS1, and KIR2DL2) displayed significant differences from corresponding genes in other Caucasoid populations. In general, Basques present an increase in activating KIR gene frequency and, consequently, the proportion of B haplotypes is higher. Three novel haplotypes were identified in the Basque population. Overall, our results confirm the particular genetic characteristics of the Basque population. No association between KIR gene content and susceptibility to T1DM was observed. PMID- 16698435 TI - Introduction: MICA/MICB in innate immunity, adaptive immunity, autoimmunity, cancer, and in the immune response to transplants. PMID- 16698434 TI - Disease relevant HLA class II alleles isolated by genotypic, haplotypic, and sequence analysis in North American Caucasians with pemphigus vulgaris. AB - Early studies of genetic susceptibility to pemphigus vulgaris (PV) showed associations between human leukocyte antigen (HLA) DR4 and DR6 and disease. The emergence of DNA sequencing techniques has implicated numerous DRB1 and DQB1 loci in various populations, leading to confusion regarding which exact alleles confer susceptibility. The strong linkage disequilibrium among DR and DQ HLA alleles further complicates the investigation of the true susceptibility loci. In this study, we report genotyping data for the largest sampling of North American Caucasian non-Jewish and Ashkenazi Jewish PV patients studied to date and compare our data with other population studies. To pinpoint true susceptibility, alleles among overrepresented sequences, we applied a step-wise reductionist analysis through (1) determination of the degree of linkage disequilibrium (LD) between purportedly associated alleles, (2) haplotype frequencies comparisons, and (3) primary sequence comparisons of disease-associated versus non-disease-associated alleles to identify crucial differences in amino acid residues in putative peptide binding pockets. Collectively, our data provide extended support for the hypothesis that the HLA associations in Caucasian PV patients map to DRB1*0402 and DQB1*0503 alone. Further structure-function studies will be required to define the exact mechanisms of HLA-mediated control of susceptibility and resistance to disease. PMID- 16698436 TI - MICA allele-level typing by sequence-based typing with computerized assignment of polymorphic sites and short tandem repeats within the transmembrane region. AB - MICA genes are located close to, but are structurally distinct from, HLA class I genes and have been found to be associated with some diseases and with immune responses to transplants. We have developed a MICA typing method based on polymerase chain reaction (PCR)/sequence-based typing and a computer program that determines the polymorphisms and distinguishes the GCT repeats in exon 5. One PCR amplification was performed to obtain templates of 2.2 kb including exons 2, 3, 4, and 5 of MICA to be sequenced with two forward and two reverse primers. Overlay of nucleotide sequencing signals resulting from presence of different GCT repeats in exon 5 antisense from two different MICA alleles can be identified by a computer-based analysis of the combined signal string containing the 35 bases. Eighteen reference samples from the 10th International Histocompatibility Workshop with known MICA alleles, as more recently determined, were tested and the concordance was 100% with the previous typing. In addition, 46 samples from kidney or heart transplant recipients and donors were analyzed for their MICA typing by this approach. Results demonstrated that the majority of samples were MICA heterozygous. The most common allele was MICA*00801/A5.1 (44.7%), which was consistent with previous reports. Three samples manifested ambiguous results, either because of polymorphism in exon 6 which was not tested or because the combination of two alleles gives the same pattern as the other two. The procedure was relatively simple and fast and is presently our method of choice for high resolution clinical MICA typing. PMID- 16698437 TI - Diversity of MICA and linkage disequilibrium with HLA-B in two North American populations. AB - The MICA gene has a high degree of polymorphism. Allelic variation of MICA may influence binding of these ligands to the NK cell receptor NKG2D and may affect organ transplantation and/or disease pathogenesis. Knowledge of the population distribution of MICA alleles and their linkage disequilibrium (LD) with class I human leukocyte antigen (HLA) will enhance our understanding of the potential functional significance of the MICA polymorphism. In the present study, we characterized the MICA and HLA-B polymorphisms in two North American populations: European and African. The individual racial groups showed rather limited variation at the MICA locus, where the same set of three most common alleles, MICA*00201, *004, and *00801, account for 64 and 71% of the allele frequency in European-Americans and African-Americans, respectively. Other common alleles (allele frequency >5% in a population) include MICA*00901 and *010. MICA alleles showed strong linkage disequilibrium with HLA-B. Typically, a common MICA allele has strong LD with several HLA-B alleles, whereas most HLA-B alleles and their related serological groups are associated with a single MICA allele. The lack of evidence for an active diversification of the MICA gene after racial separation indicates an evolutionary history distinct from that of the classical HLA genes. PMID- 16698438 TI - Regulation of NKG2D ligand gene expression. AB - The activating immunoreceptor NKG2D has seven known host ligands encoded by the MHC class I chain-related MIC and ULBP/RAET genes. Why there is such diversity of NKG2D ligands is not known but one hypothesis is that they are differentially expressed in different tissues in response to different stresses. To explore this, we compared expression patterns and promoters of NKG2D ligand genes. ULBP/RAET genes were transcribed independent of each other in a panel of cell lines. ULBP/RAET gene expression was upregulated on infection with human cytomegalovirus; however, a clinical strain, Toledo, induced expression more slowly than did a laboratory strain, AD169. ULBP4/RAET1E was not induced by infection with either strain. To investigate the mechanisms behind the similarities and differences in NKG2D ligand gene expression a comparative sequence analysis of NKG2D ligand gene putative promoter regions was conducted. Sequence alignments demonstrated that there was significant sequence diversity; however, one region of high similarity between most of the genes is evident. This region contains a number of potential transcription factor binding sites, including those involved in shock responses and sites for retinoic acid-induced factors. Promoters of some NKG2D ligand genes are polymorphic and several sequence alterations in these alleles abolished putative transcription factor binding. PMID- 16698439 TI - Intracellular expression of MICA in activated CD4 T lymphocytes and protection from NK cell-mediated MICA-dependent cytotoxicity. AB - MICA is a stress-regulated molecule recognized by the NK cell-activating receptor NKG2D. Previously, we demonstrated that MICA is induced on activated T cells but regulation by mitogenic cytokines and its biological consequences remain unexplored. Here, we show that IL-2, IL-4, and IL-15 but not TNF-alpha or IFN alpha induced MICA expression in T lymphocytes present in peripheral blood mononuclear cells (PBMCs), as assessed by Western blot. IL-2 effect involved Jak3/STAT5, p38 MAPK, p70(56) kinase, Lck/fyn kinases, and NF-kappaB. MICA expression was also observed in Th1 and Th2 cells. However, surface expression was not detected. T lymphocytes present in PBMCs and isolated CD4+ T lymphocytes stimulated with phorbol-12-myristate-13-acetate and ionomycin also induced MICA expression as assessed by Western blot, but only low levels were expressed at the cell surface. Activated but not resting CD4+ T lymphocytes were lysed by IL-15- or IL-2-stimulated NK cells, and susceptibility was increased when HLA class I molecules were blocked. Also, cytokine-stimulated NK cells produced more IFN gamma after culture with activated CD4+ T lymphocytes. However, the participation of MICA in these responses, if any, was marginal. Confocal microscopy revealed that MICA is retained mostly inside activated CD4+ T cells. Our results suggest that low surface expression of MICA on activated CD4+ T lymphocytes might be a safeguard mechanism to protect them from NK cells in an inflammatory, virus infected, or tumor microenvironment, where NK and activated CD4+ T cells are recruited. PMID- 16698440 TI - Contact inhibition causes strong downregulation of expression of MICA in human fibroblasts and decreased NK cell killing. AB - The polymorphic MICA gene encodes glycoproteins that activate T cells and NK cells through the NKG2D receptor and may costimulate immune functions. We found that MICA was expressed on freshly isolated human fibroblasts and was markedly decreased when fibroblasts were grown to confluency in culture dishes. MICA surface protein was measured by flow cytometry with the MICA-specific monoclonal antibody (mAb) 6B3, and HLA class I-specific protein was determined with mAb w6/32. In these experiments, after culture for 120 hours, the staining for MICA in fibroblasts decreased to about 20% of the initial amount and MICA mRNA fell in parallel, while HLA class I staining was maintained or even became somewhat stronger. In other experiments, MICA expression was not decreased when fibroblast contact was prevented by the addition of 1 muM Rottlerin, a specific inhibitor of protein kinase C delta known to prevent contact inhibition of fibroblasts. In the NK cell cytotoxicity assay, blocking MICA by antibody or downregulation by cell contact resulted in a decrease of specific killing by 30%. Increased MICA expression during proliferation of fibroblasts may support the host response to injury. PMID- 16698441 TI - Release of MICB molecules by tumor cells: mechanism and soluble MICB in sera of cancer patients. AB - MICA, a ligand of the activating immunoreceptor NKG2D, is released by tumor cells in a soluble form and can be detected in sera of tumor patients at significant levels. Soluble MICA has been proposed to counteract NKG2D-mediated immunosurveillance of tumors. Here, we report that MICB, the second member of the human MIC protein family, is likewise shed by metalloproteases from tumor cells and is present in sera of patients with gastrointestinal tumors. While cell-bound MICB causes downregulation of surface NKG2D, soluble MICB did not alter NKG2D expression on NK cells in vitro. Thus, proteolytic shedding of MICB by tumor cells may impair immunogenicity of tumors primarily by reducing NKG2D-ligand densities on malignant cells. PMID- 16698442 TI - MHC class I chain-related gene a diversity in head and neck squamous cell carcinoma. AB - Many immune-related genes are located within the human leukocyte antigen (HLA) region on chromosome 6. The MHC class I chain-related gene A (MICA), located centromeric of HLA-B, is involved in the innate and adaptive immune response through activation of NK and T cells. Differences of MICA transmembrane repeat lengths have been associated with diseases and expression is observed on epithelial tumors. Head and neck squamous cell carcinoma (HNSCC) is an epithelial tumor. In the present study we evaluated the MICA repeat length diversity in relation to MICA expression in Dutch HNSCC patients. MICA short tandem repeat analysis indicated a significant decrease in the frequency for the MICA-A9 repeat in patients diagnosed with oral cavity squamous cell carcinoma (SCC) but not in patients with SCC in the hypoharynx, larynx, or oropharynx. Interestingly, the majority of patients expressed MICA as observed with immunohistochemical staining whereas no soluble MICA was detected in patients' sera by enzyme-linked immunosorbent assay. In conclusion, the length of the MICA transmembrane repeats in Dutch HNSCC patients does not influence the MICA expression on tumor cells. PMID- 16698443 TI - How NKG2D ligands trigger autoimmunity? AB - The function of NK and CD8 T cells in the elimination of infected, transformed, or stressed cells occurs together with tolerance to self, a property that is essential to prevent autoimmunity. Inappropriate expression of NK receptor ligands, leading to activation of autoreactive effector cells, might therefore trigger or exacerbate autoimmunity. We review here some recent data on the activating receptor NKG2D and its MIC ligand, which are indicative of their detrimental roles in some autoimmune disorders. PMID- 16698444 TI - MHC class I chain-related gene B promoter polymorphisms and celiac disease. AB - The possibility that susceptibility to celiac disease (CD) might be influenced by the MHC class I chain-related gene family, MICA and MICB, has been previously reported. In this study, we analyzed the MICB promoter and examined the association of the polymorphisms found within such in a group of CD patients. To study the MICB promoter we sequenced the 5' flanking region of MICB gene in DNA from homozygous B-lymphoblastoid cell lines corresponding to the most frequent MICB alleles found in our population (MICB*00502, MICB*002, MICB*004, and MICB*008). DNA from a MICB*003 homozygous individual was also analyzed. Sequence analysis revealed six single nucleotide polymorphisms located at positions 45860 C/A, 45862 G/C, 45877 C/G, 46113 A/C, 46219 G/C, and 46286 G/C and an insertion of 2 bp --/AG at position 45944 according to the published genomic sequence. Those polymorphisms were found to be associated in four different haplotypes corresponding to different MICB alleles. Subsequently, 126 CD subjects and 117 healthy controls were typed by polymerase chain reaction using sequence-specific primers for these polymorphisms. MICB promoter polymorphism haplotypes were also found in our population and showed strong linkage disequilibrium with MICB alleles. MICB promoter polymorphism Haplotype 3, included in MICB*002 and MICB*008 alleles, was found to be overrepresented in CD patients (79.4% CD patients vs 45.3% healthy controls; p(c) < 0.0001; OR = 4.64; CI 95% = 2.64 8.16). Both MICB*008 and MICB*002 alleles were found as part of the CD susceptibility extended haplotypes B8/DR3/DQ2, B18/DR3/DQ2, and DR4/DQ8. PMID- 16698445 TI - MICA antigens stimulate T cell proliferation and cell-mediated cytotoxicity. AB - In previous experiments we have found that transplant recipients had specific antibodies against MICA. In the present study, we measured T cell proliferation, cytokine production, and cytotoxicity to investigate whether immunization with MICA can produce a specific cellular immune response. BALB/c mice were immunized with recombinant MICA (rMICA). Lymphoid cell suspensions obtained after immunization were used to measure T cell proliferation. We detected a robust proliferative response in MICA-stimulated cultures as determined by [3H]thymidine uptake. Using carboxyfluorescein diacetate succinimidyl ester (CFSE) to measure proliferation, we found that in MICA-stimulated cultures, 21% of the CD3+ T cells were CD4+ and CFSE-low and 3% of the T cells were proliferating CD8+ T cells. Among CFSE-low CD4+ spleen cells, 25% secreted IL-4 and only 2% produced IFN gamma, suggesting a predominant Th2-type response. Blocking of MHC class I or class II molecules with monoclonal antibodies resulted in prominent inhibition of CD8+ or CD4+ T cell proliferation, respectively. In addition, we found that blocking the NKG2D receptor did not cause inhibition of the T cell response. MICA stimulated CD8+ T lymphocytes exerted cytotoxicity against a BALB/c monocyte cell line (RAW 267.4) primed with soluble rMICA. Our results suggested that MICA activated T cells may have a role in a cellular component of transplant rejection. PMID- 16698446 TI - Frequency of MIC antibody in rejected renal transplant patients without HLA antibody. AB - Antibodies to MICA and MICB antigens were sought in the sera of 139 kidney transplant recipients. MICA*001, *002, *007, *008, and MICB*002 antigens were produced in Escherichia coli and then tested using enzyme-linked immunosorbent assay plates. Among 35 normal sera, 6% had MIC antibodies, and among 14 sera from pregnant women, 21% had MIC antibodies. Among 34 patients with functioning transplants with human leukocyte antigen (HLA) antibodies, 24% had MIC antibodies, and 19% of 32 patients without HLA antibodies had MIC antibodies. Among 46 patients who lost grafts with HLA antibodies, 26% had MIC antibodies, and among 27 failed patients without HLA antibodies, 37% had MIC antibodies. We conclude that antibodies to MIC are produced in the course of immunization by pregnancies and kidney transplants. They also occurred more frequently in rejected patients (30%) than in those with functioning grafts (21%). PMID- 16698447 TI - Detection of anti-MICA antibodies in patients awaiting kidney transplantation, during the post-transplant course, and in eluates from rejected kidney allografts by Luminex flow cytometry. AB - Previously we have reported on the development of antibodies against MICA alleles in kidney transplant recipients. These alloantibodies have now been determined using a new assay using Luminex beads bound to soluble recombinant MICA antigens produced in insect cells. In the present study we have analyzed sera from 85 kidney transplant recipients on the waiting list and 66 patients transplanted within the last 4 years and 59 acid eluates obtained from allograft nephrectomy specimens. Many of the patients in those groups were sensitized and some had previous transplants (waiting list: 15%; post-tx: 7.6%; eluates 22%) and their sera were found to contain anti-human leukocyte antigen (HLA) and anti-MICA antibodies. Anti-MICA antibodies were detected in 21/85 (24.7%) of the waiting list patients and in 15/66 (22.7%) of the transplanted recipients; 11 of the eluates (18.6%) were found to have MICA-specific antibodies (6 of them also had anti-HLA antibodies and 5 did not). These data suggested that immunization against mismatched MICA alleles induces development of anti-MICA antibodies. The finding of MICA allele-specific antibodies in eluates of kidney transplants suggests that anti-MICA antibodies can be involved in the pathogenesis of kidney allograft rejection. Further studies will be required to determine whether patients who produce alloantibodies against MICA alleles are at risk for transplant rejection even when no HLA antibodies are detected. PMID- 16698448 TI - MICA, MICB, and MHC beta block matching in bone marrow transplantation: relevance to transplantation outcome. AB - Genetic testing of the MHC is required for selection of donors for bone marrow transplantation. The outcome of related bone marrow transplantation is usually superior to that of unrelated bone marrow transplantation. This may be the result of inaccurate or incomplete genetic testing employed for selection of donor for transplantation. Based on MHC haplotype matching, MHC block matching has been developed for selection of potential unrelated donor. Block matching has been shown previously to improve outcome when added to the conventional method of human leukocyte antigen (HLA) typing for selection of donors. In this study, we have retrospectively analyzed 44 donor recipient pairs from the Australian Bone Marrow Donor Registry Repository with respect to matching of HLA-B and HLA-Cw by sequence-based typing and MICA and MICB by polymerase chain reaction-sequence specific primer and MHC beta block matching and correlated these results with survival. Beta block matching was correlated with MIC matching (p < 0.005) and with HLA-B and HLA-Cw matching. Patients who were HLA-B and -Cw matched had significantly improved survival when they were additionally matched for MHC beta block (p(c) = 0.04) or MIC (p(c) = 0.05). PMID- 16698449 TI - A critical analysis of nonsurgical treatment of Peyronie's disease. AB - OBJECTIVE: Because the efficacy of nonsurgical therapy of Peyronie's disease is controversial, this review analyses the current status of conservative therapy of Peyronie's disease. METHOD: A systematic survey on results of studies published as original papers in peer-reviewed journals is provided. RESULTS: Oral drug therapies include potassium para-aminobenzoate (Potaba), vitamin E, colchicine, tamoxifen, propoleum, acetyl-L-carnitine, and propionyl-L-carnitine. Verapamil, interferon-alpha2a and interferon-alpha2b, collagenase, cortisone, hyaluronidase, and superoxide dismutase are considered intralesional therapies that have had various degrees of success. Other treatments include local gels, iontophoresis, extracorporeal shock wave therapy, and radiation. CONCLUSION: This review analyses the current status of the conservative therapy of Peyronie's disease, because the efficacy of the nonsurgical therapy is controversial. PMID- 16698450 TI - Classification of excimer laser profiles. PMID- 16698451 TI - Imaging quality of intraocular lenses. PMID- 16698453 TI - Macular thickness after cataract surgery with intracameral cefuroxime. PMID- 16698455 TI - Association of IFIS and vasodepressor medication. PMID- 16698456 TI - Biaxial phacoemulsification. PMID- 16698457 TI - Association between IFIS and Flomax. PMID- 16698458 TI - Intraoperative retinoscopy. PMID- 16698459 TI - Silicone lens fracture using IOL injector. PMID- 16698461 TI - Consultation section. Cataract surgical problem. PMID- 16698470 TI - Technique for measuring laser in situ keratomileusis flap thickness using the IntraLase laser. AB - Measurement of laser in situ keratomileusis flap thickness is required to estimate residual bed thickness. Historically, flap thickness has been assumed, estimated based on microkeratome head labeling, guessed based on flap handling characteristics, or calculated by subtraction pachymetry. We describe a method for measuring flap thickness before actual flap elevation using the IntraLase laser. PMID- 16698471 TI - Modified SST-USST for tamsulosin-associated intraoperative [corrected] floppy iris syndrome. AB - I describe a technique using ophthalmic viscosurgical devices to perform cataract surgery in patients taking tamsulosin (Flomax). The 6-step method uses a combination variant of the soft-shell and ultimate soft-shell techniques and involves adjustments to flow parameters. It achieves satisfactory iris stability and permits uneventful surgery. PMID- 16698472 TI - Surgical technique for anterior segment surgery in pediatric patients using 25 gauge instruments. AB - Many surgical techniques for the management of vitreous prolapse in the wound, secondary pupillary membrane, and congenital cataract in pediatric patients have been described. We report our experience using 25-gauge instruments and describe intraoperative and postoperative advantages. PMID- 16698473 TI - Surgically induced astigmatism after phacoemulsification in eyes with mild to moderate corneal astigmatism: temporal versus on-axis clear corneal incisions. AB - PURPOSE: To determine whether there is a difference in surgically induced astigmatism (SIA) and postoperative uncorrected visual acuity (UCVA) and best corrected visual acuity (BCVA) between 2 types of clear corneal incisions used in phacoemulsification: the temporal and the on-axis (ie, on the steeper corneal meridian) clear corneal incision (CCTI and CCOI, respectively). SETTING: Moorfields Eye Hospital, London, United Kingdom. METHODS: In a prospective randomized controlled trial (pilot study), 61 eyes with cataract and mild to moderate corneal astigmatism (<2.58 diopters [D] on corneal topography) having phacoemulsification (single surgeon, 3.2 mm incision) were randomized to receive CCTI or CCOI. Main outcome measures included postoperative BCVA and corneal astigmatism and SIA (calculated using the Holladay vector analysis formula). Measurements were repeated postoperatively at 3 weeks in all eyes, whereas only 46 eyes made it to the final assessment at 8 weeks. Continuous variables between groups were compared by Student t test. The power of the study was calculated. RESULTS: At the first follow-up, the difference in SIA between the 2 incision types was 0.15 D and it was not statistically significant (0.65 D in CCOI; 0.50 D in CCTI). At the second follow-up, the difference was 0.29 D and it was statistically significant (0.63 D in CCOI; 0.34 D in CCTI; P = .0004). There was no statistically significant difference in the final postoperative visual acuity. The power of the pilot study was 72%. CONCLUSION: Seven weeks after small incision phacoemulsification, the CCTI induced less SIA than the CCOI; however, there were no significant differences in the final UCVA and BCVA. PMID- 16698474 TI - Change in intraocular pressure within 1 week of phacoemulsification and intraocular lens implantation using Adatocel. AB - PURPOSE: To examine the change in intraocular pressure (IOP) within 1 week of phacoemulsification and foldable posterior chamber intraocular lens (PC IOL) implantation using Adatocel (hydroxypropyl methylcellulose 2% [HPMC]). SETTING: Department of Ophthalmology, University of Sciences, Faculty of Medicine, Pecs, Hungary. METHODS: In this prospective study, the IOP in 118 eyes of 118 patients (57 men, 61 women, mean age 68 years +/- 7.8 [SD]) with no history of glaucoma was assessed by Goldmann applanation tonometry 2 to 3, 6 to 8, and 22 to 24 hours and 1 week after uneventful phacoemulsification and PC IOL implantation. The effect of the removal of Adatocel ("partial removal" from the anterior chamber [AC] only versus "complete removal" from behind of the IOL as well), the lens type (Medicontur 601 HP versus Bausch & Lomb Hydroview), and the type of anesthesia (topical versus parabulbar) were compared. Statistical analysis was performed using the Student t test, and P< or =.05 was considered statistically significant. RESULTS: The mean preoperative IOP was 13.83 +/- 2.5 mmHg. There were no significant differences at any time in postoperative IOP measurements between the 2 IOL types and the 2 modes of anesthesia. At 2 to 3 hours, 6 to 8 hours, and 22 to 24 hours, the IOP was significantly higher in the 30 eyes in which the Adatocel was partially removed (from the AC only) than in the 88 eyes in which it was completely removed (from behind the PC IOL as well) (P< or =.05, P< or =.01, and P< or =.001, respectively). CONCLUSION: Severe postoperative IOP spikes in nonglaucomatous patients after uneventful phacoemulsification cataract surgery are rare. The type of implanted PC IOL and the mode of anesthesia had no significant effect on postoperative IOP. Total removal of the ophthalmic viscosurgical device, even when using HPMCs such as Adatocel, is necessary to prevent postoperative IOP spikes. PMID- 16698475 TI - Visual experience and patient satisfaction with wavefront-guided laser in situ keratomileusis. AB - PURPOSE: To determine the correlation between patient perception and clinical measurements after wavefront-guided laser in situ keratomileusis (LASIK). SETTING: Multicenter clinical trial. METHODS: In this retrospective analysis of clinical study results in 274 eyes, wavefront-guided LASIK procedures were performed in 274 myopic-astigmatic eyes at 6 sites. Comprehensive evaluations of vision and ocular health were conducted preoperatively and for 6 months postoperatively. Visual acuity and contrast sensitivity were measured, and questionnaires were administered. Questionnaire responses were compared with clinical measurements. RESULTS: The overall distribution shifted toward "more satisfied." The "very satisfied" population increased by as much as 70% in all areas. There was no significant change in frequency of visual symptoms. Patients were more expressive about visual decline than visual improvement. Perception of changes in vision appeared to be related to mesopic contrast sensitivity and mesopic contrast sensitivity under glare. Mesopic pupil diameter was not a major predictor of patient satisfaction. Contact lens wearers were more satisfied with postoperative vision than were spectacle wearers. Men were more likely to report visual symptoms and to complain about night vision, although their changes were the same as those of the women. Residual refractive error was a major predictor for most questionnaire items. CONCLUSION: Most patients were as satisfied or more satisfied with their postoperative uncorrected visual acuity than with their preoperative best corrected visual acuity. By continuing to minimize postoperative refractive error through the use of nomograms and improved technology, visual complaints such as night glare and halo could be diminished or even eliminated. The study shows that subjective experience affected satisfaction with results. Some dissatisfaction might be mitigated by being sure patients are educated and have realistic expectations before surgery. PMID- 16698476 TI - Q-factor customized ablation profile for the correction of myopic astigmatism. AB - PURPOSE: To compare the results of the Q-factor customized aspheric ablation profile with the wavefront-guided customized ablation pattern for the correction of myopic astigmatism. SETTING: Institute for Refractive and Ophthalmic Surgery, Zurich, Switzerland. METHODS: Thirty-five patients were enrolled in a controlled study in which the nondominant eye was treated with the Q-factor customized profile (custom-Q study group) and the dominant eye was treated with wavefront guided customized ablation (control group). Preoperative and 1-month postoperative high-contrast visual acuity, low-contrast visual acuity, and glare visual acuity, as well as aberrometry and asphericity of the cornea, were compared between the 2 groups. All eyes received laser in situ keratomileusis surgery, and the laser treatment was accomplished with the Wavelight Eye-Q 400 Hz excimer laser. RESULTS: For corrections up to -9 diopters (D) of myopia, there were no statistically significant differences between the 2 groups regarding any visual or optical parameter except coma-like aberrations (3rd Zernike order), where the wavefront-guided group was significantly better 1 month after surgery (P = .002). For corrections up to -5 D (spherical equivalent), the Q-factor optimized treated eyes had a significantly smaller shift toward oblate cornea: DeltaQ15 = 0.25 in Q-factor customized versus DeltaQ15 = 0.38 in wavefront-guided treatment (P = .04). CONCLUSIONS: Regarding safety and refractive efficacy, custom-Q ablation profiles were clinically equivalent to wavefront-guided profiles in corrections of myopia up to -9 D and astigmatism up to 2.5 D. Corneal asphericity was less impaired by the custom-Q treatment up to -5 D of myopia. PMID- 16698477 TI - Laser in situ keratomileusis: ablation on the flap and stromal bed in a primary treatment. AB - PURPOSE: To analyze outcomes of laser in situ keratomileusis (LASIK) performed on the flap and on the stromal bed at the same time in a primary treatment. SETTING: Instituto de la Vision, Buenos Aires, Argentina. METHODS: Twenty-two cases (13 female, 9 male) were studied retrospectively; the mean age of the patients was 33.15 years +/- 10.44 (SD). Inclusion criteria were low pachymetry or topographic asymmetry. Twenty cases (90.9%) presented with low pachymetry (512.2 +/- 21.40 mum), and 2 cases (9.1%) presented with topographic asymmetry (inferior-superior difference more than 1.5 diopters [D]). Ablation was performed on the flap from the back and on the bed. The flap was made using the Technolas Hansatome microkeratome and the ablation by the Technolas 217z excimer laser (both by Bausch & Lomb). Preoperatively, the spherical equivalent (SE) was -5.61 +/- 1.72 D, the cylinder was -1.78 +/- 1.24 D, and the best spectacle-corrected visual acuity (BSCVA) was 0.77 +/- 0.19. Difficulties and complications of the technique, efficacy and safety index, and aberrations were analyzed. RESULTS: According to the surgical plan and our nomogram, diopters corrected on the flap were -1.73 +/- 1.08 and on the bed were -4.77 +/- 1.89. The mean follow-up was 8.28 +/- 2.19 months. At 6 months, the SE was -0.21 +/- 0.39, vectorial change was 1.69 +/- 0.74, and the BSCVA was 0.77 +/- 0.20. Postoperatively, no eye had an increase in refractive astigmatism. Uncorrected visual acuity was 20/40 or better in all cases (100%) and 20/25 or better in 13 cases (59.1%). Regarding gained and lost lines of BSCVA, 54.5% conserved the lines, 18.2% lost 1 line, 9.1% gained 1 line, and 18.2% gained 2 lines. CONCLUSIONS: Simultaneous ablation on the flap and on the bed in cases of low pachymetry or topographical asymmetry was predictable, effective, and safe. PMID- 16698478 TI - Outcome of simultaneous phakic implantable contact lens removal with cataract extraction and pseudophakic intraocular lens implantation. AB - PURPOSE: To assess the outcome of simultaneous implantable contact lens (ICL) removal and cataract extraction with pseudophakic intraocular lens (IOL) implantation. SETTING: CODET Aris Vision Institute, Tijuana, Mexico. METHODS: This retrospective noncomparative interventional case series evaluated 14 eyes of 12 patients with ICL implantations who developed a cataract and simultaneously had ICL removal and cataract extraction with IOL implantation. The follow-up time was at least 6 months (range 6 to 24 months). Visual acuity (logMAR), manifest refraction, intraocular pressure, and adverse events were recorded. RESULTS: Of the 12 patients (14 eyes), 10 patients (12 eyes) had ICL surgery to correct high myopia and 2 patients (2 eyes), to correct hyperopia. The mean uncorrected visual acuity after ICL implantation (before cataract development), before cataract surgery, and after cataract surgery were 0.48 +/- 0.32, 0.83 +/- 0.34, and 0.40 +/- 0.27, respectively. The mean best corrected visual acuity (BCVA) before ICL implantation, after ICL implantation, and after cataract surgery were 0.31 +/- 0.21, 0.28 +/- 0.19, and 0.27 +/- 0.21, respectively. The mean final manifest spherical equivalent was 0.30 diopters (D) +/- 1.07 (SD) (range +2.38 to 2.0 D). Ten eyes (71.4%) were within +/-1.0 D of the calculated target. One eye had a tear in the posterior capsule with vitreous loss during cataract surgery. No other intraoperative, perioperative, or postoperative complications were observed. No loss of BCVA was recorded at the last postoperative visit. CONCLUSIONS: Lens opacities and cataract formation are a potential complication of ICL surgery. The removal of the ICL and the cataract with IOL implantation was found to be safe, with predictable refractive results. PMID- 16698479 TI - Analysis of crystalline lens position. AB - PURPOSE: To study normal crystalline lens position to provide a comparative baseline for future studies of crystalline lens or intraocular lens shift. SETTING: Taipei Municipal Yang-Ming Hospital, Taipei, Taiwan. METHODS: A Scheimpflug anterior segment analyzer (EAS-1000, Nidek) was prospectively applied to measure the cycloplegic crystalline lens position in subjects who had not had previous ocular surgeries or who had been diagnosed previously with major ocular diseases such as glaucoma, retinal detachment, or cataract. Measurements included anterior chamber depth (ACD), magnitudes, direction of lens decentration, and lens tilt. Refractive error was measured with an autorefractometer, and multiple linear regression was used to verify revealed relationships. The aging effect was determined with the Pearson correlation test. RESULTS: Thirty-nine eyes of 30 subjects (15 men, median age 13 years, range 4 to 53 years) were included. The center of the anterior lens surface was decentered 0.25 mm superotemporally. The lens tilted 2.85 degrees with the anterior lens surface facing the inferotemporal quadrant. The mean ACD was 3.26 mm; it tended to increase before subjects reached 20 years of age and to decrease thereafter. With age, the lens tended to exhibit less tilt. Lens position did not affect the spherical equivalent or the magnitude of astigmatism. CONCLUSION: The crystalline lens was not aligned perfectly along the visual axis, but its effect on refraction was limited. PMID- 16698480 TI - Subjective difficulty of each stage of phacoemulsification cataract surgery performed by basic surgical trainees. AB - PURPOSE: To measure the relative difficulty of each stage of phacoemulsification surgery performed by basic surgical trainees. SETTING: Mater Misericordiae University Hospital, Dublin, Ireland. METHODS: This prospective study comprised 100 consecutive cases of phacoemulsification performed by 8 trainee surgeons over an 11-month period. The trainees completed a questionnaire at the end of surgery, rating the degree of difficulty of 8 individual stages of phacoemulsification on a 5-point scale from 1 (very easy) to 5 (very difficult). RESULTS: At the start of the study, trainees had spent a mean of 9.4 months training. The mean preoperative best corrected decimal visual acuity was 20/70. The most difficult stages were phacoemulsification and capsulorhexis with the highest mean scores of 3.32 +/- 1.00 (SD) and 3.30 +/- 1.24, respectively and the lowest completion rates of 66.7% and 74.4%, respectively. The first 3 stages of peribulbar blockade (1.72 +/- 0.75), draping the surgical field (1.35 +/- 0.70) and corneal incision (1.97 +/- 0.72) were easiest and had close to a 100% completion rate. Hydrodissection, irrigation/aspiration, and intraocular lens insertion had difficulty rates of 2.42 +/- 0.92, 2.80 +/- 1.10, and 2.58 +/- 1.08, respectively, and completion rates of 92.3%, 78.4%, and 83.1%, respectively. Posterior capsule tear occurred in 9%, including 4% with vitreous loss; there was 1 case of zonular dehiscence. CONCLUSIONS: Trainee surgeons found phacoemulsification and capsulorhexis the most difficult stages of cataract surgery early in the learning curve. More time can be dedicated to mastering these steps in the wet lab. PMID- 16698481 TI - Early post-phacoemulsification hypotony as a risk factor for intraocular contamination: in vivo model. AB - PURPOSE: To observe cataract wound dynamics and determine the risk for intraocular contamination under induced conditions of early postoperative hypotony after anterior chamber decompression for high intraocular pressure (IOP) spikes after uneventful phacoemulsification surgery. SETTING: Department of Ophthalmology, Queen's Hospital, Burton-on-Trent, United Kingdom. METHODS: In a prospective case series, 30 patients were treated with anterior chamber decompression for elevated IOP (>35 mm Hg) 1 to 2 hours after uneventful phacoemulsification through clear corneal temporal incisions. A drop of 2% fluorescein was instilled in the conjunctival sac before aqueous release to study the ocular fluid movements during and after the decompression procedure. The patterns of fluorescein movement were observed and photographed under cobalt blue light. RESULTS: Fluorescein staining of the entire corneal tunnel was observed in all 30 eyes after decompression. An immediate fluorescein flare was observed in the anterior chamber after aqueous release in 24 eyes (80%) (P<.001). Observations over several seconds after decompression found a frank influx of fluorescein-stained fluid into the anterior chamber in 12 eyes (40%) (P<.001), giving rise to an "inverse Seidel's test." An intermittent ingress of fluorescein stained fluid continued for several minutes after the procedure in 6 eyes (20%) (P<.05). CONCLUSIONS: An incompetence of clear corneal cataract wound with intraocular contamination was observed during the period of induced hypotony after anterior chamber decompression in the early post-phacoemulsification period. PMID- 16698482 TI - Characterizing pseudoexfoliation syndrome through the use of ultrasound biomicroscopy. AB - PURPOSE: To determine the clinical utility of ultrasound biomicroscopy (UBM) in diagnosis of pseudoexfoliation (PEX) syndrome by characterizing the lens capsule and zonules before cataract surgery. SETTING: Veterans Administration Hospital, East Orange, and University of Medicine & Dentistry of New Jersey-New Jersey Medical School, Newark, New Jersey, USA. METHODS: Ultrasound biomicroscopy was performed on 10 patients clinically diagnosed with PEX syndrome. The clinical diagnosis was made by the presence of fibrillin deposits on the anterior lens capsule, lack of pigment at the pupillary ruff, and poor pharmacologic dilation. Five persons without PEX were used as controls. The thickness of the anterior lens capsule was measured in 5 locations in each eye: centrally and in the peripheral lens capsule superiorly, inferiorly, nasally, and temporally. Four measurements were taken from the zonule at the thickest point on each fiber. The UBM also found the presence or absence of nodular deposits on the zonules. RESULTS: The anterior and peripheral lens capsule in patients with PEX was thicker than that in the control group. Additionally, patients with PEX had thicker zonules than the control group and had nodular deposits present; the control group had no deposits. These differences were all significant with a 99% confidence interval. CONCLUSIONS: A thicker anterior lens capsule and lens zonule nodules were associated with PEX. These abnormalities can be visualized with the UBM to confirm the diagnosis of PEX and identify patients at risk for operative complications. PMID- 16698483 TI - Superoxide dismutase activity in the lens capsule of patients with pseudoexfoliation syndrome and cataract. AB - PURPOSE: To evaluate the activity of the antioxidative enzyme superoxide dismutase (SOD) in the anterior lens capsule of patients with cataract complicating pseudoexfoliation syndrome (PEX) in an attempt to determine the role of the oxidative mechanisms in the etiopathogenesis of this disorder. SETTING: Departments of Ophthalmology and Biochemistry, Ankara University School of Medicine, Ankara, Turkey. METHODS: At the beginning of phacoemulsification surgery, the anterior lens capsules in 21 eyes with PEX and cataract (group A) and those in 24 eyes with cataract alone (group B) were collected with continuous curvilinear capsulorhexis and were frozen at -20 degrees C. At the time of analysis, the specimens were thawed and centrifuged and the supernatants of the homogenate obtained from the samples were analyzed for the activity of the enzyme SOD. RESULTS: The mean age of patients was 74.8 years +/- 5.5 (SD) in group A and 71.7 +/- 5.6 years in group B. The specific activity of SOD was 17.6 +/- 10.8 IU/mg and 4.36 +/- 1.80 IU/mL in group A and 9.9 +/- 12.9 IU/mg and 2.71 +/- 2.61 IU/mL in group B. Superoxide dismutase activity was significantly higher in group A patients than in group B patients (P = .022 and P = .011, respectively). CONCLUSIONS: The increase in SOD activity in the lens capsule of patients with PEX and cataract suggests that oxidative mechanisms play a role in the etiopathogenesis of cataract in PEX. This suggests that SOD activity may be increased as a compensatory mechanism to eliminate this oxidative stress. PMID- 16698484 TI - Effect and outcomes of posterior capsule rupture in a district general hospital setting. AB - PURPOSE: To assess how patients from a district general hospital population were affected on a short-term basis after posterior capsule (PC) rupture during phacoemulsification cataract surgery. SETTING: Department of Ophthalmology, Raigmore Hospital, Inverness, United Kingdom. METHODS: This retrospective audit encompassing a 2-year period evaluated the incidence of PC rupture and its outcomes. Details of the patients' age, preoperative and final postoperative best corrected visual acuity (BCVA), ocular comorbidity, surgeon grade, number and duration of follow-up reviews, and postoperative complications were collected. RESULTS: Over the 2-year period, the PC rupture rate was 1.7% (45 PC ruptures in 2727 phacoemulsification cataract procedures). Thirty-eight eyes (84.4%) achieved a final BCVA of 6/12 or better. In 39 eyes (86.7%), final BCVA improved compared with BCVA at presentation. Mean duration of follow-up was 11.7 weeks. The most common extra procedure was anterior vitrectomy (64.4%). The most common postoperative complication was raised intraocular pressure (IOP) (exceeding 30 mm Hg) on the first postoperative day (20.0%) despite additional acetazolamide as prophylaxis. None developed infective endophthalmitis. Apart from 1 retinal detachment (2.2%) at week 27, all complications resolved by 19 weeks. No patient required additional long-term medication at the final visit. CONCLUSION: Posterior capsule rupture impacted on the patient in the short-term in terms of additional surgical procedures, additional topical and oral medications, and number and duration of follow-up reviews. Most patients recovered an acuity of 6/12 or better. However, it is important to be vigilant of postoperative complications, especially raised IOP on the first postoperative day. PMID- 16698485 TI - Visual outcomes after accommodating intraocular lens implantation. AB - PURPOSE: To evaluate and compare the visual outcomes and accommodative amplitude in cataract patients after implantation of the Crystalens intraocular lens (IOL) (Eyeonics) versus standard monofocal IOLs. SETTING: Ten clinics in a nationwide multicenter study in the United States. METHODS: A multicenter comparative interventional case series with masked randomized postoperative examination of 224 eyes of 112 patients was performed by a single observer. Patients were divided into 2 groups (56 patients; 112 eyes each) depending on which IOL was implanted (Crystalens or monofocal). Accommodation was measured using 1 objective (dynamic retinoscopy) and 2 subjective methods (defocus and near point of accommodation). Visual acuity measurements were performed under the same conditions with standard visual acuity charts. RESULTS: Uncorrected monocular near vision was significantly better in the Crystalens group than in the standard monofocal group, with 101 of 112 eyes (90%) and 17 of 112 (15%), respectfully, reading J3 or better postoperatively. All 56 Crystalens patients had a binocular uncorrected near visual acuity of J3 or better compared with 16 of 56 (29%) standard monofocal patients. The mean postoperative monocular (0.85 +/- 0.30 [SD] versus 0.70 +/- 0.19, P<.01) and binocular (1.16 +/- 0.17 versus 1.01 +/- 0.14, P<.01) distance uncorrected visual acuities were also better in the Crystalens group than in the control group. All patients in the study achieved a corrected distance visual acuity of 20/20 or better. Measures of accommodation were significantly higher in Crystalens patients than in the monofocal IOL patients (dynamic retinoscopy 2.42 +/- 0.39 diopters [D] versus 0.91 +/- 0.24 D, P<.01; monocular defocus 1.74 +/- 0.48 D versus 0.75 +/- 0.25 D, P<.01; monocular near point of accommodation 9.5 +/- 3.1 inches versus 34.7 +/- 9.8 inches, P<.01). Perceived accommodation (5.79 D) was significantly greater than the measured accommodation (1.96 to 2.42 D) in Crystalens patients (paired t test, P<.01). CONCLUSIONS: The Crystalens IOL provided better uncorrected near and distance visual outcomes than standard monofocal IOLs in all analyses performed. Patients perceived a greater accommodation than measured. Understanding why this occurred could lead to valuable advances in accommodating IOL technology. PMID- 16698486 TI - Posterior capsule opacification in myopic eyes. AB - PURPOSE: To determine whether posterior capsule opacification (PCO) is extensive in eyes with myopia or long axial length when an intraocular lens (IOL) of low power, zero-power, or minus-power, is implanted. SETTING: Hayashi Eye Hospital, Fukuoka, Japan. METHODS: Ninety eyes of 90 patients scheduled for phacoemulsification surgery were recruited. These consisted of 30 eyes with high (> or =-8 diopters [D]) myopia, 30 eyes with moderate (<-8 D and > or =-3 D) myopia, and 30 eyes with low (<-3 D) myopia. All eyes had implantation of an acrylic IOL-low-power, zero-power, or minus-power-with a sharp optic edge (AcrySof MA60BM or MA60MA, Alcon Surgical). The PCO in these eyes was measured using a Scheimpflug videophotography system (EAS-1000, Nidek) 1, 3, 6, 12, 18, and 24 months after surgery. Visual acuity and the incidence of neodymium:YAG (Nd:YAG) laser posterior capsulotomy were also examined. RESULTS: No significant difference was observed in the mean PCO value or in the Nd:YAG capsulotomy rate between the high myopia, moderate myopia, or low myopia groups throughout the follow-up period. There was also no significant correlation between PCO value and the actual spherical power or axial length of the eye. Furthermore, although mean visual acuity tended to be worse in proportion to the degree of myopia, the difference was not statistically significant. CONCLUSION: When an acrylic IOL of low-power, zero-power, or minus-power with a sharp optic edge was implanted, high myopia and long axial length were not associated with the degree of PCO. PMID- 16698487 TI - Thermal effect of microburst and hyperpulse settings during sleeveless bimanual phacoemulsification with advanced power modulations. AB - PURPOSE: To assess wound temperature during bimanual sleeveless phacoemulsification using customizable power modulations such as hyperpulse and microburst technology. SETTING: In vitro laboratory. METHODS: The Millennium Microsurgical System (Bausch & Lomb) with custom control software (CCS) was used to perform phacoemulsification in 5 porcine eyes with MicroFlow needles (Bausch & Lomb) and with power varied from 20% to 80% in 10% increments. Pulse modes were set for fixed microburst (4 ms on, 4 ms off; and 6 ms on, 12 or 24 ms off) and for hyperpulse (30% duty cycle with 8 or 75 pulses per second [pps]), with and without aspiration-line occlusion. Wound temperatures were measured 3 times per second. RESULTS: Using 80% total power, the wound temperature during 3 minutes of occlusion did not exceed 39.0 degrees C. The maximum temperature with fixed microbursts of 4 ms on, 4 ms off was 29.0 degrees C without occlusion and 37.8 degrees C with occlusion (duration 3 minutes). At 6 ms on, 12 ms off, the maximum temperatures were 28.1 degrees C and 38.7 degrees C, respectively. At 6 ms on, 24 ms off, peak temperatures were 24 degrees C and 23.6 degrees C, respectively. The hyperpulse mode of 30% duty cycle and 8 pps produced maximum temperatures of 25.5 degrees C nonoccluded and 33.4 degrees C occluded. With 30% duty cycle, 75 pps, temperatures were 28 degrees C and 38.0 degrees C, respectively. For all power below 80%, temperatures were lower. CONCLUSIONS: Customizable power modulation with microburst and hyperpulse technology further reduced wound temperatures during bimanual sleeveless phacoemulsification. This enhances the safety and effectiveness of phacoemulsification through a sleeveless needle and a small stab incision. PMID- 16698488 TI - Optomechanical eye model with imaging capabilities for objective evaluation of intraocular lenses. AB - PURPOSE: To develop an in vitro procedure providing data on the visual performance obtainable with intraocular lenses (IOLs), for objective comparison between IOL models and direct correlation with the relative visual performance attainable in vivo. SETTING: University Hospital San Raffaele, Milan, Italy. METHODS: An optomechanical eye model was developed to allow simulated in vivo testing of IOLs. The experimental eye mimics the optics and geometry of the Gullstrand's eye model, with an aspheric poly(methyl methacrylate) cornea, variable pupil, and IOL holder. Its detection system is designed to reproduce the mean resolution of the human fovea. The imaging capabilities of the model eye were measured using monofocal IOLs. The tests included qualitative information, such as appearance of optotype chart images, and quantitative information, such as simulated visual acuity tests for far and near distance at variable contrasts. RESULTS: Objective numerical IOL evaluation was made possible on the basis of the visual acuity recorded with the eye model. The maximum recorded far acuity for the monofocal IOLs was about 20/14 at full contrast, progressively decreasing for reduced contrast. Best corrected near acuity ranged between 20/14.7 and 20/15.4. CONCLUSIONS: The optomechanical eye model provided objective grading of IOLs through the evaluation of simulated visual acuity, which can be scaled usefully to human vision. The eye model also allowed the qualitative visualization of IOL imaging properties, making it potentially useful in characterizing and distinguishing different IOL types. PMID- 16698489 TI - Hydrophilic acrylic intraocular lens as a drug-delivery system: Pilot study. AB - PURPOSE: To evaluate the ability of a hydrophilic acrylic intraocular lens (IOL) to serve as a drug-delivery system for commercially available gatifloxacin and moxifloxacin. SETTING: David J. Apple, MD, Laboratories for Ophthalmic Devices Research, John A. Moran Eye Center, University of Utah, Salt Lake City, Utah, USA. METHODS: Hydrophilic acrylic IOLs (C-flex, Rayner Ltd.), presoaked for 24 hours in commercially available solutions of gatifloxacin (Zymar) or moxifloxacin (Vigamox), were implanted in the capsular bag of 6 rabbits for a total of 12 eyes (6 in each group). Aqueous humor samples were taken 2, 4, and 6 hours after implantation. One rabbit served as a control and had nonpresoaked C-flex IOLs implanted. At the end of the operation, 1 drop of Vigamox was applied to the right eye and 1 drop of Zymar was applied to the left eye of the control rabbit. RESULTS: High concentrations of both antibiotics were found in all the samples of the eyes implanted with the presoaked IOLs. The concentrations of the antibiotics decreased over time, but even the 6-hour sample concentrations were markedly higher than the concentrations found in the control rabbit after 4 hours. CONCLUSION: The results suggest that the Rayner C-flex IOL can be effective as a drug-delivery system for fourth-generation fluoroquinolones. PMID- 16698490 TI - Interlenticular opacification: dual-optic versus piggyback intraocular lenses. AB - PURPOSE: To evaluate and compare the incidence of capsular bag opacification, particularly interlenticular opacification (ILO), in rabbit eyes implanted with a dual-optic silicone intraocular lens (IOL) or piggyback lenses. SETTING: John A. Moran Eye Center, University of Utah, Salt Lake City, Utah, USA. METHODS: Ten dual-optic study IOLs (Synchrony), 10 control pairs of piggyback silicone-plate lenses, and 10 control pairs of piggyback single-piece hydrophobic acrylic lenses were implanted in the capsular bag of 30 rabbit eyes following phacoemulsification. After a 6-week follow-up, the rabbits were killed and their eyes enucleated. Anterior capsule opacification and posterior capsule opacification were graded on a 0 to 4 scale from a posterior or Miyake-Apple view. Interlenticular opacification was noted in relation to the center of the interlenticular space (periphery, paracentral, and central area) and to the number of quadrants involved. The eyes were then evaluated histopathologically. RESULTS: Postoperative inflammatory reaction was similar in all groups. Interlenticular opacification formation was statistically different among the 3 groups of lenses (ILO extension, P = .0013, and ILO extension x ILO quadrants, P = .0023; Kruskal-Wallis test). Pairwise post comparisons of ILO formation showed that the differences between the study IOL group and the silicone-plate lens group were not significant. Interlenticular opacification post comparisons between the hydrophobic acrylic lenses and the study lens or the silicone-plate lenses were significant (P = .002 and P = .001, respectively). Histopathologic examination showed extension of the proliferating cortical material from the peripheral Soemmering's ring into the interlenticular space, causing ILO, especially with the pairs of hydrophobic acrylic lenses. CONCLUSIONS: In this rabbit model, ILO was significantly associated with pairs of hydrophobic acrylic lenses implanted in the bag. This study appears to confirm clinical observations that implantation of 2 silicone-plate lenses in the bag is not associated with ILO. There was also a relative lack of ILO with the dual-optic silicone lens. PMID- 16698491 TI - Intraocular lens haptic fracturing with the neodymium:YAG laser In vitro study. AB - PURPOSE: To determine the neodymium:YAG (Nd:YAG) laser energy threshold to fracture the haptic of various currently marketed intraocular lenses. SETTING: Dokuz Eylul University, Ophthalmology Department, Izmir, Turkey. METHODS: Four haptic materials-poly(methyl methacrylate) (PMMA), hydrophobic acrylic, hydrophilic acrylic, and polyvinylidene fluoride-were tested. The intraocular lenses (IOLs) were grouped according to haptic material. Each group comprised 9 IOLs. Beginning laser energy was 4 mJ. If no crack was noted, energy was increased by 0.5 mJ at every 20 shots and the procedure continued until a fracture was noted. If the fracture could not be developed despite reaching an energy level of 10 mJ, the procedure was terminated. Statistical analysis was performed to determine the differences within groups. RESULTS: Hydrophobic acrylic and polyvinylidene fluoride haptics required higher energy and more laser shots to create a fracture than hydrophilic acrylic and PMMA haptics. CONCLUSIONS: Neodymium:YAG laser haptic fracturing can be a viable option to cut the fibrosed haptic before explanting any IOL. It was demonstrated in vitro that it is easier to fracture hydrophilic acrylic and PMMA haptics than hydrophobic acrylic and polyvinylidene fluoride haptics. PMID- 16698492 TI - Effect of surface coating an acrylic intraocular lens with poly(2 methacryloyloxyethyl phosphorylcholine) polymer on lens epithelial cell line behavior. AB - PURPOSE: To evaluate the effect of surface coating of an acrylic intraocular lens (IOL) with poly(2-methacryloyloxyethyl phosphorylcholine) (MPC) on the behavior of the lens epithelial cell (LEC) line, alpha-TN4. SETTING: Department of Ophthalmology, Nihon University School of Medicine, Tokyo, Japan. METHODS: A hydrophobic soft acrylic IOL (AF-1, Hoya) was coated with MPC polymer. A noncoated IOL served as control. An IOL from each group was placed on the membrane of collagen I or IV of the cell culture dish. The alpha-TN4 cells were seeded in the insert. Cell behaviors (ie, cell proliferation and spreading) on IOLs and membranes were observed. Cell migration beneath the IOL optic portion was assayed using a computer software program (POCOman system) for posterior capsule opacification (PCO). Type I or IV collagen is the major matrix component of PCO or native lens capsule. RESULTS: Cell proliferation was more marked on the noncoated IOL than on the coated IOL. Type IV collagen accelerated proliferation more than type I collagen. Cell migration to the area beneath the IOL optic was more prominent in the group with the type I collagen membrane and noncoated IOL than in other groups. CONCLUSION: Coating an acrylic IOL surface with MPC polymer suppressed adhesion and proliferation of LECs, suggesting it improves IOL biocompatibility. PMID- 16698493 TI - Late spontaneous in-the-bag intraocular lens and capsular tension ring dislocation in pseudoexfoliation syndrome. AB - Two patients with pseudoexfoliation syndrome (PEX) in whom late spontaneous in the-bag intraocular lens (IOL) and capsular tension ring (CTR) dislocation occurred 3 and 6 years after cataract surgery are described. The patients received CTRs because of phacodonesis due to zonular laxity. The IOLs were centered postoperatively, and there was no postoperative ocular trauma. In both cases, removal of the IOL and CTR within the capsular bag was performed uneventfully by a scleral tunnel incision and an anterior chamber IOL was implanted. Capsular tension ring implantation in PEX-associated zonular weakening does not guarantee long-term zonular stability and capsular bag/IOL position in these patients after cataract surgery. PMID- 16698494 TI - Keratitis after intracorneal ring segment insertion for keratoconus. AB - We report a case of sterile keratitis after Ferrara intracorneal ring (Ferrara Ophthalmics) implantation into the right cornea of a keratoconus patient who also suffered from atopic blepharoconjunctivitis. The keratitis resolved within a few days after removal of the Ferrara rings. Ocular atopy with sterile keratitis should be considered in the differential diagnosis of keratitis after the insertion of intracorneal ring segments, which can be easily treated by ring removal. PMID- 16698495 TI - Management of corneal ectasia and cataract following photorefractive keratectomy. AB - A 42-year-old man was referred to our clinic 18 months after bilateral photorefractive keratectomy (PRK). He had been on topical prednisolone acetate for 12 months because of post-PRK grade 4 haze. On his first visit, visual acuity was limited to light perception in both eyes because of moderate haze, significant corneal ectasia, and a white cataract. A 2-step surgical approach was elected in both eyes. First, a deep anterior lamellar keratoplasty was performed. Six weeks later, phacoemulsification with intraocular lens implantation was performed. Compared with a triple procedure combining penetrating keratoplasty and cataract surgery in 1 stage, the 2-step approach may lower the risk for corneal graft rejection and reduce ametropia. PMID- 16698497 TI - Severe photic phenomenon. AB - A 69-year-old man who had uneventful bilateral cataract surgery with visual acuity of 20/15 without correction presented with severe dysphotopsia. Treatment with brimonidine (Alphagan) or pilocarpine did not resolve the symptoms. A multipiece silicone intraocular lens was piggybacked into the sulcus, and the patient no longer reported dysphotopsia. PMID- 16698496 TI - Exophiala dermatitidis keratitis after laser in situ keratomileusis. AB - A 52-year-old man had laser in situ keratomileusis in March 2002 and 2 weeks later had a corneal infection that was found to be caused by Exophiala dermatitidis. This infection waxed and waned for 9 months despite therapy with natamycin, itraconazole, and fluconazole but eventually responded to topical amphotericin. He recovered good visual acuity in this eye with a rigid gas permeable lens. This case shows the importance of selecting the appropriate agent in fungal keratitis. PMID- 16698498 TI - Phacoemulsification in a patient with a deep brain stimulator. AB - We describe a 62-year-old man with a deep brain stimulator who had cataract surgery performed by phacoemulsification. Representatives of Medtronic and Alcon Laboratories would not sanction our proceeding with cataract surgery because of the possible interaction between the ultrasound and the device. Simulated cataract surgery 1 week before the scheduled operation failed to produce any consequences. One week later, the patient had uneventful cataract surgery. Although a single case does not guarantee the lack of interference between the ultrasound used in phacoemulsification and the deep brain stimulator, our safe outcome may warrant a place in the ophthalmic literature. PMID- 16698499 TI - Paradoxical increase of visual impairment with punctal occlusion in a patient with mild dry eye. AB - We report a paradoxical increase in visual impairment after punctal plug placement despite improvement in corneal epithelial damage caused by mild dry eye. Ocular higher-order aberrations (HOAs) were measured sequentially with a wavefront sensor before and after punctal plug insertion in a patient with mild dry eye. Although postblink HOAs tend to increase in normal subjects or patients with dry eye, after treatment there was a maximum spike in HOAs for a few seconds that decreased gradually. Excessive retention of tear film by punctal occlusion may cause inferosuperior asymmetrical distribution in precorneal tear-film thickness, possibly leading to slower wavefront on the inferior cornea and increased coma-like aberrations. Increased visual impairment might occur paradoxically despite improvement in the corneal epithelial damage caused by dry eye. PMID- 16698500 TI - Reduction of anterior-segment vascularity with preoperative topical apraclonidine 1%. PMID- 16698501 TI - Can patients self-assess their requirement for cataract surgery? PMID- 16698502 TI - The circle and the shadow: 2 early postoperative findings. PMID- 16698503 TI - Clinical trials update from the American College of Cardiology: Darbepoetin alfa, ASTEROID, UNIVERSE, paediatric carvedilol, UNLOAD and ICELAND. AB - This article provides information and a commentary on trials relevant to the pathophysiology, prevention and treatment of heart failure, presented at the American College of Cardiology 55th Annual Scientific Session held in March 2006. All reports should be considered as preliminary data, as analyses may change in the final publication. Darbepoetin alfa increased haemoglobin levels in heart failure patients and improved some aspects of quality of life compared to placebo. In the ASTEROID study rosuvastatin significantly reduced LDL-cholesterol levels and induced regression of atherosclerosis in patients with CAD. Rosuvastatin also produced a significant reduction in LDL-cholesterol levels in heart failure patients in the UNIVERSE study, but had no effect on left ventricular remodelling compared to placebo. The paediatric carvedilol study failed to show a benefit of carvedilol in children with heart failure. Ultrafiltration produced a greater weight and fluid loss than intravenous diuretics in heart failure patients with volume overload in the UNLOAD study but did not exert a greater improvement in breathlessness; however, ultrafiltration did reduce readmission rates. The ICELAND MI study showed that CMR imaging was more sensitive than ECG or clinical criteria for detecting myocardial infarction. PMID- 16698504 TI - Iowa nursing home characteristics associated with reported abuse. AB - OBJECTIVE: To evaluate the association of nursing home abuse reported by employees in Iowa licensed nursing homes with nursing home characteristics, inspection results, nursing staff hours, and census demographics. DESIGN: Mailed survey. SETTING: Iowa's 409 Medicare-certified stand-alone nursing homes. PARTICIPANTS: Administrators and directors of nursing. RESULTS: The annual rate of abusive events was 20.7 per 1000 nursing home residents, with a rate of 18.4 reported events and 5.2 substantiated events. A higher number of residents and a higher number of certified beds were significantly associated with higher incident, report, and substantiation rates. Higher incident and report rates were associated with nursing homes in metropolitan areas. Higher substantiation rates were associated with for-profit nursing homes. CONCLUSION: Recognized abuse in nursing homes is not uncommon. Approximately 90% of events are reported to the state authorities with 29% of those being substantiated. Differences associated with nursing facility abuse rates are metropolitan area, ownership, occupancy rate, and number of residents and certified beds. PMID- 16698505 TI - Predictive role of single diseases and their combination on recovery of balance and gait in disabled elderly patients. AB - OBJECTIVES: In the elderly population, chronic diseases are common determinants of mobility limitations and comorbidity consistently shows a strong association with functional status. This study was designed to evaluate the role of single chronic diseases and of their combination on functional recovery after rehabilitative treatment in disabled elderly patients. DESIGN: With respect to the difference in magnitude of their disabling effect, diseases were classified into 2 groups: "more disabling" diseases (COPD, heart failure, peripheral artery diseases, diabetes, and not life-threatening cancer) and "less disabling" diseases (anemia, kidney, gastrointestinal, and liver diseases). SETTING: 35-bed Geriatric Evaluation and Rehabilitation Unit. PARTICIPANTS: We studied 710 patients (age 77.8 +/- 7.4 years, 76.2% females), consecutively admitted for stroke, Parkinson's disease, and osteoarthritis. MEASUREMENTS: A multidimensional evaluation for mobility (Tinetti-score), cognitive status (MMSE), and somatic health (Greenfield's Individual Disease Severity Index-IDS, Burden of diseases BoD) was performed. Functional recovery was decided based on the Delta-Tinetti, which is the difference of the values between admission and discharge. RESULTS: We tested, in a multivariate regression model, the predictive role of single chronic conditions and of their combinations on functional recovery, after having adjusted for which diseases are direct causes of disability (stroke, Parkinson's disease, and osteoarthritis) and other potential predictors (age, sex, cognitive function, depressive symptoms, albumin, and c-reactive protein). A negative prediction of functional recovery was expressed by the "more disabling" diseases group. The determinants of poor recovery were characterized by the combination of "more disabling diseases" rather than single condition effects, independently by age, cognitive, and functional status on admission. CONCLUSION: Our study adds a new perspective about the role of COPD, heart failure, peripheral artery diseases, diabetes and not life-threatening cancer on functional recovery, emphasizing their combined impact in elderly people. PMID- 16698506 TI - The accuracy of Minimum Data Set diagnoses in describing recent hospitalization at acute care facilities. AB - OBJECTIVES: The Minimum Data Set (MDS) is the resident assessment instrument used to guide clinical care, reimbursement, and assess quality in long-term care facilities. This database has been used in many studies, although the accuracy of many data elements remains unknown. This study evaluated the accuracy of the MDS diagnosis variables with respect to the diagnoses for recent hospitalization from Medicare claims data. DESIGN: Retrospective cohort study. SETTING: 945 skilled nursing facilities in Ohio. PARTICIPANTS: 17,294 residents admitted from an acute care facility during 2000. MEASUREMENTS: Eleven diagnoses listed in the MDS were compared with Medicare hospital discharge claims. Specifically, each MDS diagnosis was compared to the primary diagnosis, the list of secondary diagnoses, and the Diagnosis Related Group (DRG). RESULTS: Claims diagnoses were listed in the MDS with an average frequency of 79% (range: 31%-94%) for the primary diagnosis, 66% (range: 33%-90%) for any diagnosis, and 71% (range: 31%-94%) for the DRG. MDS diagnoses were listed as the primary diagnosis, any diagnosis, and DRG with an average frequency of 20% (range: 6%-81%), 62% (range: 41%-86%), and 19% (range: 7%-84%), respectively, with only hip fracture listed more than 80% of the time. CONCLUSION: The sensitivity of the MDS for listing diagnoses from recent hospitalization appears good for most diagnoses. However, except for hip fracture, the MDS has poor predictive value with regard to the primary reason for the preceding hospitalization; this may have implications for resident care planning and the utility of this database in long-term care research. PMID- 16698507 TI - Symptom experiences of residents dying in assisted living. AB - OBJECTIVES: To identify the end-of-life symptom experiences of residents who died in assisted living, to describe family satisfaction with the end-of-life care, and to compare end-of-life symptom distress across 4 settings: assisted living, private home, nursing home, and hospital. DESIGN: In-person interviews with family members of people aged 65 or older who had died 2 to 4 months earlier in an assisted-living facility. Symptom distress reports were compared with data from prior studies in other settings. SETTING: A mixed urban-rural community in the Northwest. PARTICIPANTS: Twenty-five family members who self-identified as closely involved in the care or decision making for a relative who died in an assisted-living facility. MEASUREMENTS: A 28-item interview guide composed of demographic questions, the modified Family Memorial Symptom Assessment Scale Global Distress Index (FMSAS-GDI), and open-ended items describing the decedent's last days and family satisfaction with care and symptom management. RESULTS: Symptoms experienced by at least 75% of decedents were lack of energy, loss of appetite, dry mouth, drowsiness, and pain. Symptom experiences in assisted living were comparable to those in other settings. Families overall were satisfied with end-of-life care in assisted living, but had concerns about communication among providers, inadequate monitoring of their decedent, and lack of staff knowledge specific to symptom management. CONCLUSION: Residents dying in assisted-living settings have similar symptom distress experiences as people dying in other, more studied settings. As residents and families alike overwhelmingly support resident deaths in their assisted living "home," policies, structures, and processes are needed to ensure that end-of-life care needs are met. PMID- 16698508 TI - The Aging Game: improving medical students' attitudes toward caring for the elderly. AB - OBJECTIVES: The objective of this study was to evaluate the effectiveness of a modified aging game to enhance medical students' attitudes toward caring for elderly patients, enhance empathy for elderly patients, and improve general attitudes toward the elderly. DESIGN: Preintervention and postintervention study PARTICIPANTS AND SETTING: All 84 students from 2 first-year medical school classes INTERVENTION: The aging game takes place over a period of 3 hours with a group of 10 to 12 students and 5 to 6 facilitators, and includes experiential learning about polypharmacy, medication cost, loss of functional status, semi independent living, and living in a long-term care facility. MEASUREMENTS: Scores obtained through a modified Maxwell and Sullivan questionnaire, and the Aging Semantic Differential (ASD). RESULTS: There was a statistically significant improvement in 6 of the 8 attitudes toward caring for elderly and a statistically significant increase in empathy towards elderly patients following the aging game. There was a statistically significant change on 23 out of 32 ASD questions. There was no evidence of differences in the scores from pre-course to post-course by gender age group, interest in Family/Internal Medicine, or prior geriatric experience. Following the game, 77 (93%) of the students said they would take the course if not mandatory (P = .0001). A majority stated the aging game added significantly (61.5%) or moderately (37.3%) to their knowledge and skills in patient care for the elderly. CONCLUSION: A simulation experience like the aging game is an effective method of increasing the empathy and attitudes toward caring for the elderly early in the medical school curriculum. Longitudinal assessment of the effect of the aging game in maintaining the positive effect on empathy and attitudes toward caring for the elderly and especially in clinical practice needs to be done. PMID- 16698509 TI - The efficacy of performing screening mammograms in the frail elderly population. PMID- 16698510 TI - The application of evidence-based principles of care in older persons (issue 3): management of diabetes mellitus. PMID- 16698512 TI - Physical abuse of the elderly: the medical director's response. PMID- 16698513 TI - Quality improvement initiative for chronic pain assessment and management in the nursing home: a pilot study. AB - OBJECTIVES: To improve assessment and management of chronic pain in the nursing home through a method of continuous quality improvement. DESIGN: Quality improvement. SETTING: One nursing home in North Carolina as the primary site and 3 related facilities as secondary sites. INTERVENTION: Formation of a quality improvement team that reviewed current clinical practice guidelines, updated the policies and procedures, developed tools for pain assessment, and educated the staff in pain assessment and management principles. The "Plan-Do-Study-Act" (PDSA) paradigm was employed. MEASUREMENT: The staff's pain-related knowledge was measured with multiple-choice tests before and after an educational program. The completeness of the documentation and satisfaction of patients and families with pain assessment and management were also measured before and after the intervention. RESULTS: Before the intervention, the mean number of questions correct on the staff test was highest among the registered nurses (RN) (13.7 out of 16), intermediate among the licensed practical nurses (LPN) (12.4), and lowest among the certified nursing assistants (CNA) (9.4). After the intervention the scores were 16 for the RNs, 12.9 for the LPNs, and 12.0 for the CNAs. In addition, 8 of 9 elements of a complete pain assessment showed improvement in documentation. The level of satisfaction for both the patients and families measured by interviews also increased. CONCLUSION: Education and use of the PDSA paradigm improved staff knowledge and patient and family satisfaction with chronic pain assessment and management. PMID- 16698514 TI - Delirium in postacute and long-term care. AB - Delirium is a classic geriatric syndrome that occurs commonly among the frail elders who make up many of the residents in postacute and long-term care facilities. The prevalence of the disorder in these settings may be increasing as a result of the pressure to reduce hospital length of stay. Clinicians often do not recognize when patients in their care are delirious, but simple and practical means exist to allow its diagnosis. Those who practice in long-term care must be knowledgeable about the risk factors for the disorder, as well as how to recognize, diagnose, prevent, and treat it. PMID- 16698515 TI - "Pay for performance": can it help improve long-term care? PMID- 16698516 TI - The race. PMID- 16698517 TI - Can combination drug therapy potentially reduce multiple medications and polypharmacy in nursing homes? PMID- 16698518 TI - Value creation: the new business of medicine. PMID- 16698519 TI - Unusual appearing leiomyoma at laparoscopic myomectomy. PMID- 16698520 TI - Laparotomy to laparoscopy: changing trends in the surgical management of ectopic pregnancy in a tertiary care teaching hospital. AB - STUDY OBJECTIVE: To review the changing trends in the surgical management of ectopic pregnancy and to evaluate the effect of an ongoing training program for resident surgeons on the rate and success of laparoscopic surgery. DESIGN: Retrospective chart review (Canadian Task Force classification II-3). SETTING: University tertiary medical center. PATIENTS: One thousand forty-six patients with ectopic pregnancy treated at Jackson Memorial Hospital from January 1, 1995, through December 31, 2004. INTERVENTION: A formal, ongoing laparoscopic training program was established in 1999. MEASUREMENTS: The rate of laparoscopy, laparotomy, and conversion for each year was compared with the baseline year of 1995 and between the years before and after the establishment of the training program. RESULTS: Four hundred sixty-eight laparotomies and 578 laparoscopies were successfully completed. The laparoscopic approach rate has increased from 40.9% in 1995 to 86.3% in 2004. By year 2000 and thereafter, significantly more ectopic surgeries were approached through the laparoscope compared with in 1995 (p<.01). By 2000 and thereafter, significantly less conversions occurred compared with the baseline year of 1995 (4.0% vs 18.5%; p = .01). CONCLUSION: Laparoscopy became the primary approach for the management of ectopic pregnancy in part because of resident participation in an ongoing laparoscopy training program. PMID- 16698521 TI - Hysteroscopic endometrial ablation using the Hydro ThermAblator in an outpatient hysteroscopy clinic: feasibility and acceptability. AB - STUDY OBJECTIVE: To assess the feasibility and patient acceptability of global endometrial ablation using the Hydro ThermAblator (HTA) in an outpatient hysteroscopy clinic setting under local anaesthesia. DESIGN: A prospective cohort study (Canadian Task Force classification II-2). SETTING: University hospital in the United Kingdom. PATIENTS: Forty patients with medication-resistant menorrhagia. INTERVENTION: Oral mefenamic acid was administered 8 hours before the procedure. A combined cervical/paracervical block using lignocaine 1% adrenaline 1:200,000 was used before hysteroscopic endometrial ablation using the HTA device. MEASUREMENTS AND MAIN RESULTS: A successful procedure was defined as a completed HTA treatment cycle. Failure was defined as premature termination or inability to carry out the HTA procedure after scheduling the patient. Pain scores were estimated at three stages during and after the procedure using a visual-analog scale. A questionnaire was used before discharge to assess acceptability. Forty patients were successfully treated. The median pain score during the ablation was 6.4 (range 4.0-8.9). Eighty-eight percent of patients found the procedure acceptable. No serious complications occurred. All patients were discharged home within 2 hours of the procedure. CONCLUSION: Selected patients suffering from menorrhagia may be treated with the HTA device using local anesthesia in an outpatient setting. PMID- 16698522 TI - Perioperative complication rate in 1706 patients after a standardized laparoscopic supracervical hysterectomy technique. AB - STUDY OBJECTIVE: The aim of the study was to determine perioperative morbidity and complication rate after a standardized technique of laparoscopic supracervical hysterectomy (LASH). DESIGN: Retrospective analysis of consecutive patients (Canadian Task Force classification II-3). SETTING: Private hospital. PATIENTS: Seventeen hundred and six consecutive patients with symptomatic uterine myomata, dysfunctional uterine bleeding, dysmenorrhea, or chronic pelvic pain. INTERVENTION: Laparoscopic supracervical hysterectomy using a unipolar hook for dissection of the body of the uterus followed by electric morcellation. MEASUREMENTS AND MAIN RESULTS: The main indications for LASH were uterine myomata with dysfunctional uterine bleeding (31.1%) or without (45.4%) and therapy resistant dysfunctional uterine bleeding (21%). The mean uterine weight was 226.4 +/- 193.9 g (95% CI 217.1-235.6), the mean duration of surgery was 91.4 +/- 33.3 minutes (95% CI 89.9-93.0), and the mean duration of hospital stay was 2.15 +/- 0.63 days (95% CI 2.12-2.18). Nine hundred two (52.9%) of the patients had a history of at least one laparotomy. In 14 patients (0.82%), a conversion to laparotomy was necessary. Of these, 11 were due to the size and immobility of the uterus, one was due to severe adhesions, and two because of intraoperative complications. In total, five (0.3%) intraoperative and 20 (1.2%) postoperative complications occurred. The mean weight of the uterus in the five patients with intraoperative complications (three bladder injuries, one ureter injury, and one severe intraoperative bleed) was 818.4 +/- 911.9 g (95% CI -313.9-1950.7). In two patients who suffered trauma to the bladder, there was a history of cesarean sections (two and three, respectively). The most common postoperative complications were bleeding from the cervix and pain caused by adhesions or postoperative infection. CONCLUSION: Laparoscopic supracervical hysterectomy is a minimally invasive surgical method that should be regarded as an alternative to all other methods of total hysterectomy in benign conditions of the uterus (uterine myomata, dysfunctional uterine bleeding, uterine adenomyosis) as it is associated with a low perioperative morbidity and a rapid period of convalescence. Of special significance is that LASH can be performed on nulliparous patients, patients who have not previously had vaginal delivery, and patients who have had previous abdominal surgery. PMID- 16698523 TI - The effect of vasopressin on local and general circulation during laparoscopic surgery. AB - STUDY OBJECTIVE: To study local and general circulation after vasopressin administration during laparoscopic surgery. DESIGN: Prospective analysis (Canadian Task Force classification II-1). SETTING: University-affiliated hospital. PATIENTS: Nine patients who underwent laparoscopic myomectomy (LM) from March through December 2003. INTERVENTION: Transvaginal ultrasound and transesophageal ultrasound were performed to evaluate blood flow of the uterine artery and the descending aorta, respectively, during laparoscopic surgery. MEASUREMENTS AND MAIN RESULTS: Nine patients who had a low number (one to two) of uterine myomas in the anterior wall underwent LM with local administration of four units of 100-fold diluted vasopressin. We monitored blood flow of the ascending branch of the uterine artery and measured the resistance index (RI) value by transvaginal ultrasound. The decreasing reaction of diastole blood flow was fast, and the median time required until disappearance of signal was 4 minutes (range 1-8 minutes). We monitored blood flow of the descending aorta by transesophageal ultrasound and compared the results with eight control patients who did not receive vasopressin. Urine volume was measured during the surgery and postoperatively. No significant difference in descending aortic blood flow volume, and intraoperative and postoperative urine volume was found between the two groups. Systolic/diastolic blood pressure tended to increase immediately after vasopressin administration, but the increase was not significant. CONCLUSIONS: Local administration of vasopressin to the uterus is a safe and effective hemostatic technique for controlling regional blood flow from the uterine artery to peripheral vessels without having a significant effect on systemic circulatory dynamics. PMID- 16698524 TI - Topical diclofenac patch for postoperative wound pain in laparoscopic gynecologic surgery: a randomized study. AB - STUDY OBJECTIVE: To compare pain management of standard analgesic and standard analgesic plus diclofenac transdermal patch in patients who undergo laparoscopic gynecologic surgery. DESIGN: Randomized prospective study (Canadian Task Force classification I). SETTING: University hospital. PATIENTS: One hundred twenty patients requiring laparoscopic surgery for gynecologic benign diseases. INTERVENTIONS: Patients were divided into two groups, one medicated with a diclofenac patch (diclofenac group) and the other medicated with standard skin medication (placebo; control group) at all incisional areas at the end of the operation. MEASUREMENTS AND MAIN RESULTS: The principal measures of outcome were pain intensity at 6, 12, and 24 hours after surgery and consumption of analgesics. The two treatment groups were comparable with respect to demographic and intraoperative characteristics. No significant difference was observed between the two groups in mean pain intensity at 6 hours after surgery. Mean pain intensity at 12 and 24 hours, respectively, after surgery was significantly lower in the diclofenac group (3.7 +/- 1.3 and 2.0 +/- 0.6) than that observed in the control group (5.7 +/- 1.9 and 4.6 +/- 0.5) (p value, respectively, .002 and <.001). Twenty-one patients (35.0%) in the diclofenac group required analgesics in the first 36 hours after the operation versus 43 patients (71.7%) in the control group (p <.001). Hospital discharge was significantly more rapid in the diclofenac group (28 +/- 5 hours vs 39 +/- 3 hours; p = .031). CONCLUSION: Diclofenac transdermal administration seems a valid help to standard analgesic treatment in postoperative pain control and could also help reduce the period of hospitalization of patients who undergo laparoscopic benign gynecologic surgery. PMID- 16698525 TI - Histopathologic findings on uterosacral ligaments in women with chronic pelvic pain and visually normal pelvis at laparoscopy. AB - STUDY OBJECTIVE: To determine the prevalence and type of microscopic findings on laparoscopically resected uterosacral ligaments in women with chronic pelvic pain and no visible pelvic disease. The effect of this procedure on the patients' level of pain also was assessed as a secondary objective. DESIGN: Prospective follow-up (Canadian Task Force classification II-2). SETTING: University affiliated hospital. PATIENTS: Twenty-seven premenopausal women undergoing diagnostic laparoscopy for chronic pelvic pain had a normal pelvis on visual inspection. All patients underwent resection and histologic evaluation of the uterosacral ligaments. Pain relief was evaluated by use of a questionnaire administered before and at 3, 6, and 12 months after surgery. MEASUREMENTS AND MAIN RESULTS: Microscopic examination identified endometriosis in 2 (7.4%), endosalpingiosis in 3 (11.1%), and inflammation in 14 (51.9%) patients. Laparoscopic uterosacral ligament resection was associated with a reduction in dysmenorrhea (p < or = .001), with 14 (52%) patients reporting improved or resolved symptoms. There was a statistically significant decrease in dyspareunia (p < or = .01) and in the severity of noncyclical pain (p < or = .002). Thirty five percent of patients no longer required medication for pain control (p < or = .005). The number of days needed off work also decreased. CONCLUSION: Despite normal laparoscopic appearance, microscopic endometriosis, endosalpingiosis, and inflammatory changes were found in uterosacral ligaments in 17 (63%) women with chronic pelvic pain. Laparoscopic resection of uterosacral ligaments improved dysmenorrhea, dyspareunia, and noncyclical pain and decreased the number of days lost from work, as well as the proportion of patients who required medication for pain control. PMID- 16698526 TI - Effect of body habitus and parity on insufflated CO2 volume at various intraabdominal pressures during laparoscopic access in women. AB - STUDY OBJECTIVE: To determine height, weight, body mass index (BMI), parity, and age effect on the volume of CO2 pneumoperitoneum during laparoscopic access in women. DESIGN: Prospective observational cohort study (Canadian Task Force classification II-1). SETTING: University-affiliated teaching hospital. PATIENTS: From January through June 2004, 100 healthy women underwent operative laparoscopic surgery consecutively by the senior author (GAV). Indications were: chronic pelvic pain (CPP, n = 66), pelvic mass (n = 7), CPP and pelvic mass (n = 4), infertility (n = 23). Median (range) height, weight, BMI, parity and age were 1.65 m (1.45-1.85 m), 70 kg (43-118 kg), 25.5 kg/m2 (17-39 kg/m2), 1.1 (0-5), and 34 years (19-58 years), respectively. INTERVENTIONS: While in supine position, patients received general endotracheal anesthesia and muscle relaxants. Pneumoperitoneum was established by reusable Veres needle. The insufflated CO2 volume was serially recorded at intraperitoneal insufflation pressures (IPIPs) of 10, 15, 20, 25, and 30 mm Hg. The primary 10-mm trocar was introduced at IPIP of 30 mm Hg. Upon entering the peritoneal cavity, the abdominal contents were visualized with the laparoscope to ensure there was no injury, and the IPIP was immediately reduced back to the operating pressure of 15 mm Hg before switching the patient to the Trendelenburg position. MEASUREMENTS AND MAIN RESULTS: The mean (SD) insufflated CO2 volumes at 10, 15, 20, 25, and 30 mm Hg were 1.7 (0.74) L, 3.1 (0.9) L, 3.96 (1.05) L, 4.42 (1.1) L and 4.72 (1.14) L, respectively. Using multivariate analysis, we demonstrated that at 20 to 30 mm Hg the insufflated CO2 volume correlated positively with the height, weight and BMI of women. Parity correlated positively at all pressures. There was no correlation with age at any pressure. CONCLUSIONS: Higher CO2 volume is required to establish appropriate pneumoperitoneum in tall, overweight, and parous women at 20 to 30 mm Hg. Setting the IPIP at 20 to 30 mm Hg before primary trocar insertion eliminates the need to monitor CO2 insufflated volume regardless of women's age, parity, and body habitus. PMID- 16698527 TI - A modified method of laparoscopic presacral neurectomy for the treatment of midline dysmenorrhea. AB - STUDY OBJECTIVE: To describe the modified surgical technique of laparoscopic presacral neurectomy. DESIGN: A prospective trial (Canadian Task Force classification II-1). SETTING: University hospital. PATIENTS: Forty-two women with severe midline dysmenorrhea. INTERVENTION: Laparoscopic modified presacral neurectomy (LMPSN). MEASUREMENTS AND MAIN RESULTS: We designed a prospective study and used a zero-to-10 pain score system to evaluate 42 patients before and after LMPSN. Before surgery, each patient had pain scores ranging from 7 to 10 and had failed medical management. Mean time required to perform LMPSN was 7.33 +/- 1.76 minutes, and most women were discharged the day after surgery. The mean postoperative pain score of 2.02 +/- 2.25 was obtained 6 months after surgery and again at 12 months. No patient had intraoperative or long-term complications. CONCLUSION: When a patient requires surgical treatment for chronic midline pelvic pain or severe dysmenorrhea, LMPSN is an alternative choice. It is relatively safe, simple to perform, and satisfactory. This modified surgical procedure appears to increasingly succeed in treatment of midline dysmenorrhea. PMID- 16698528 TI - Novel application of polypropylene sling: transvaginal cervicoisthmic cerclage in women with high risk of preterm delivery. AB - STUDY OBJECTIVE: To describe a novel technique of transvaginal cervicoisthmic cerclage with a polypropylene sling in prevention of preterm labor in pregnant women at high risk. DESIGN: Retrospective study (Canadian Task Force classification: III). SETTING: University hospital (department of obstetrics and gynecology). PATIENTS: Twenty-one women showing symptoms of high risk of preterm delivery: histories of pregnancy losses in the second trimester and prior failure of MacDonald's cerclage or absent portio vaginalis of the cervix. The median age of the patients was 32.8 years (range 22-39 years). INTERVENTIONS: Cerclage was performed between 12 and 16 weeks' gestation. A polypropylene tape was placed at the cervicoisthmic junction by vaginal route. Caesarean delivery was systematically performed in all patients because the cerclage was considered to be definitive. MEASUREMENTS AND MAIN RESULTS: No intraoperative complications occurred. The mean operating time was 36 +/- 6 minutes (range 30-45 minutes). The mean length of stay was 1.4 +/- 0.5 days. Mean gestational age and birth weight at delivery were respectively 37.1 +/- 1.8 weeks (CI 95%: 36.4-37.9) and 2850 +/- 745 g (CI 95%: 2531-3168). The preterm birth rate was 19% (4/21). One neonatal death occurred after amniotic fluid infection at 34 weeks. Birth at less than 32 weeks occurred in one patient (4%). CONCLUSION: Transvaginal cervicoisthmic cerclage with polypropylene sling may be considered as an effective and minimally invasive alternative to the transabdominal cervicoisthmic cerclage in women presenting with high risk of preterm delivery. PMID- 16698529 TI - Hysteroscopy for the diagnosis and treatment of pathologic changes in the uterine cavity in women with an intact hymen. AB - STUDY OBJECTIVE: We perform hysteroscopy in 14 women for the diagnosis and treatment of pathologic changes in the uterine cavity of women with an intact hymen. All hysteroscopies were successfully carried out with the hymen intact and the therapeutic results were excellent. DESIGN: Retrospective study. SETTING: University-affiliated hospital, department of obstetrics and gynecology. PATIENTS: Fourteen unmarried women who underwent hysteroscopy on the condition of keeping the hymen intact. INTERVENTIONS: Hysteroscopy was used for the diagnosis and treatment of pathological changes in the uterine cavity of women with an intact hymen. MEASUREMENTS AND MAIN RESULTS: All 14 hysteroscopies were successfully carried out with the hymen intact, and the therapeutic results were excellent. CONCLUSION: Hysteroscopy without damaging the intact hymen is feasible and helpful for the diagnosis in the treatment of pathologic endometrial changes in women. PMID- 16698530 TI - Three-dimensional model for gas flow, resistance, and leakage-dependent nominal pressure maintenance of different laparoscopic insufflators. AB - STUDY OBJECTIVE: To compare laparoscopic insufflators with different insufflation characteristics and evaluate the effects of resistance and leakage on maintenance of nominal pressure. DESIGN: Prospective study (Canadian Task Force classification II-2). SETTING: Laboratory of university clinic. INTERVENTIONS: Measurement in an abdomen box model of insufflator response to different resistance and leakage combinations during steady state while attempting to maintain a nominal set pressure of 12 mm Hg. MEASUREMENTS AND MAIN RESULTS: An abdomen box model for laboratory measurements was designed with differing entrance and exit diameters simulated by hole disks of 0.5 mm to 7.6 mm. With a computer-based data-acquisition model, five insufflators (Olympus 9L and 16L, Storz 10L and 30L, HiTec 16L) were evaluated with 150 disk combinations. Average abdominal box pressures were recorded in three-dimensional (3-D) profiles showing different nominal pressure maintenance response for all insufflators regarding the resistance and leakage combination, maximum flow performance, and insufflation principle of the device. Average abdomen model box pressure can increase up to 26.57 mm Hg at 12 mm Hg nominal pressure setting (+121.4%). Insufflators with high flow rates (Storz 30L) tend to compensate leakage better but with the risk of higher intraabdominal pressure peaks. Insufflators with low pressure principle (HiTec 16L) have limited pressure compensation but are safer for the patient. CONCLUSIONS: Nominal pressure maintenance of insufflators is mainly dependent on flow resistance and leakage rate and less on potential insufflator flow capacity. With this 3-D model, the nominal pressure maintenance of different insufflators can be described for the first time regarding these main factors. PMID- 16698531 TI - Severe complications caused by retained tissue in laparoscopic supracervical hysterectomy. AB - In our department, laparoscopic supracervical hysterectomy has gradually replaced hysterectomy by laparotomy in patients with a benign condition and no history of cervical dysplasia. During the last year, we experienced serious complications in two patients because of inadequate removal of the tissue in the morcellator procedure. These cases are presented and discussed in this case report. PMID- 16698532 TI - Sister Mary Joseph's nodule as the first presenting sign of primary fallopian tube adenocarcinoma. AB - Umbilical metastasis (Sister Mary Joseph's nodule) is often the first sign of intraabdominal and/or pelvic carcinoma. We describe the fourth case reported in the literature of Sister Mary Joseph's nodule originating from fallopian tube carcinoma. In a 54-year-old woman, Sister Mary Joseph's nodule was unexpectedly detected during umbilical hernia repair. Subsequent laparoscopy revealed a 2-cm friable tumor located at the fimbriated end of right fallopian tube and 1-cm peritoneal implant in the pouch of Douglas. Laparoscopic bilateral adnexectomy and resection of the peritoneal implant were performed. Because frozen section examination revealed fallopian tube carcinoma, the procedure was continued with laparotomy including total abdominal hysterectomy, omentectomy, and pelvic lymph node sampling. Final diagnosis was stage IIIB fallopian tube carcinoma. The patient received postoperative adjuvant chemotherapy with single-agent carboplatin and has remained alive and with no evidence of disease. It is concluded that in cases of Sister Mary Joseph's nodule, laparoscopy can be a useful tool in the search of the primary tumor in the abdomen and/or pelvis. Laparoscopy can provide crucial information with respect to the location, size, and feasibility of optimal surgical resection of the intraabdominal and/or pelvic tumors. PMID- 16698533 TI - Laparoscopic management of vaginal clear cell adenocarcinoma arising in pelvic endometriosis: case report and literature review. AB - Vaginal clear cell adenocarcinoma arising from pelvic endometriosis has not been reported in the literature. We report a case of a 50-year-old woman with stage I clear cell adenocarcinoma of the vagina who was found to have endometriosis adjacent to the vaginal tumor. She was treated with neoadjuvant chemoradiation, laparoscopically assisted radical vaginal hysterectomy, radical upper vaginectomy, and pelvic lymphadenectomy followed by combination chemotherapy. PMID- 16698534 TI - Subserosal pregnancy in a previous myomectomy site: a variant of intramural pregnancy. AB - A 35-year-old woman with a history of myomectomy underwent in vitro fertilization and became pregnant. Transvaginal ultrasound revealed a gestational sac within the subserosal area of the posterior uterine wall. The patient was treated successfully with conservative surgery, and the pathologic evaluation of the excised mass demonstrated chorionic villi involving myometrium. Early in a subsequent pregnancy, placental invasion through the sinus tract was detected. However, the pregnancy outcome was uneventful. This constitutes the first report of subserosal implantation in the uterine body. Our findings suggest that the probable pathogenesis of this rare variant of intramural pregnancy is implantation through a sinus tract made during a previous uterine surgery. PMID- 16698535 TI - Combined transurethral and laparoscopic partial cystectomy and robot-assisted bladder repair for the treatment of bladder endometrioma. AB - A 40-year-old nulliparous woman with a 7-year history of progressive abdominal discomfort and lower urinary tract symptoms was diagnosed with a deeply infiltrating 4-cm bladder endometrioma. After failing conservative therapy, she was treated with simultaneous transurethral and laparoscopic partial cystectomy and robot-assisted bladder reconstruction. She remained symptom free at last follow-up. PMID- 16698536 TI - The ABCs of a safer laparoscopic entry. AB - It is well known that at least 50% of laparoscopic complications occur during the initial entry into the abdomen regardless of the method used. There is evidence that most gynecologists practice the "classic" or closed laparoscopic entry. There is no evidence that the closed entry is more or less dangerous than the other existing methods of entry. Entry-related complications have been minimized by the following three steps: low initial Veres intraperitoneal pressure indicating correct placement of the Veres needle; transient high-pressure pneumoperitoneum before primary trocar/cannula insertion; and visual entry with the Ternamian cannula. Following the above steps, no entry complications have been encountered by the author in more than 3000 consecutive laparoscopies. PMID- 16698540 TI - Molecular glue to cement a phage. PMID- 16698538 TI - Fanfani F, et al. A prospective study of laparoscopy versus minilaparotomy in the treatment of uterine myomas. PMID- 16698541 TI - Broken symmetry in homing endonucleases. PMID- 16698542 TI - Turning up the HEAT on translation. PMID- 16698543 TI - Deposition diseases and 3D domain swapping. AB - Protein aggregation is a feature of both normal cellular assemblies and pathological protein depositions. Although the limited order of aggregates has often impeded their structural characterization, 3D domain swapping has been implicated in the formation of several protein aggregates. Here, we review known structures displaying 3D domain swapping in the context of amyloid and related fibrils, prion proteins, and macroscopic aggregates, and we discuss the possible involvement of domain swapping in protein deposition diseases. PMID- 16698544 TI - RNA kink-turns as molecular elbows: hydration, cation binding, and large-scale dynamics. AB - The presence of Kink-turns (Kt) at key functional sites in the ribosome (e.g., A site finger and L7/L12 stalk) suggests that some Kink-turns can confer flexibility on RNA protuberances that regulate the traversal of tRNAs during translocation. Explicit solvent molecular dynamics demonstrates that Kink-turns can act as flexible molecular elbows. Kink-turns are associated with a unique network of long-residency static and dynamical hydration sites that is intimately involved in modulating their conformational dynamics. An implicit solvent conformational search confirms the flexibility of Kink-turns around their X-ray geometries and identifies a second low-energy region with open structures that could correspond to Kink-turn geometries seen in solution experiments. An extended simulation of Kt-42 with the factor binding site (helices 43 and 44) shows that the local Kt-42 elbow-like motion fully propagates beyond the Kink turn, and that there is no other comparably flexible site in this rRNA region. Kink-turns could mediate large-scale adjustments of distant RNA segments. PMID- 16698545 TI - Highly discriminatory binding of capsid-cementing proteins in bacteriophage L. AB - Cementing proteins that bind to the virion surface have been described in double stranded DNA viruses such as herpesvirus, adenovirus, and numerous bacteriophages. The three-dimensional structure of bacteriophage L determined by electron cryo-microscopy reveals binding modes of two cementing proteins-one, called Dec, encoded by phage gene orf134 and the other by an as yet unidentified gene. These two proteins form homotrimers and bind at the quasi 3-fold axes nearest the icosahedral 2-fold axes and at the icosahedral 3-fold vertices, respectively. They do not bind at the quasi 3-fold axes near the icosahedral 5 fold vertices. These observations indicate precise recognition of the two cementing proteins at a subset of the quasi equivalent sites on the phage capsid. Sequence analysis shows striking similarity between the C-terminal portion of phage L Dec protein and five regions in the long tail fiber of a T4-like phage, suggesting functional parallelism between them. PMID- 16698546 TI - High-resolution structural validation of the computational redesign of human U1A protein. AB - Achieving atomic-level resolution in the computational design of a protein structure remains a challenging problem despite recent progress. Rigorous experimental tests are needed to improve protein design algorithms, yet studies of the structure and dynamics of computationally designed proteins are very few. The NMR structure and backbone dynamics of a redesigned protein of 96 amino acids are compared here with the design target, human U1A protein. We demonstrate that the redesigned protein reproduces the target structure to within the uncertainty of the NMR coordinates, even as 65 out of 96 amino acids were simultaneously changed by purely computational methods. The dynamics of the backbone of the redesigned protein also mirror those of human U1A, suggesting that the protein design algorithm captures the shape of the potential energy landscape in addition to the local energy minimum. PMID- 16698547 TI - YfhJ, a molecular adaptor in iron-sulfur cluster formation or a frataxin-like protein? AB - The yfhJ gene is part of the isc operon, which encodes the machinery devoted to assemble iron-sulfur clusters in prokaryotes. Its transcript is a small acidic protein that binds the desulfurase IscS, which is essential in iron-specific metabolic pathways. To understand its cellular role, we have characterized the structure of YfhJ in solution and its interactions with potential cellular partners. It contains a modified winged helix motif, usually present in DNA binding proteins, and is able to bind iron cations. The IscS interaction surface is the same as that involved in iron binding. This observation and the pattern of conservation through species strongly suggest that YfhJ is a molecular adaptor that is able to modulate the function of IscS in iron-sulfur cluster formation. The remarkable similarity between the in vitro behavior of YfhJ and that of the protein frataxin also suggests new hypotheses regarding the functional role of both proteins. PMID- 16698548 TI - The structure of I-CeuI homing endonuclease: Evolving asymmetric DNA recognition from a symmetric protein scaffold. AB - Homing endonucleases are highly specific catalysts of DNA strand breaks, leading to the transfer of mobile intervening sequences containing the endonuclease ORF. We have determined the structure and DNA recognition behavior of I-CeuI, a homodimeric LAGLIDADG endonuclease from Chlamydomonas eugametos. This symmetric endonuclease displays unique structural elaborations on its core enzyme fold, and it preferentially cleaves a highly asymmetric target site. This latter property represents an early step, prior to gene fusion, in the generation of asymmetric DNA binding platforms from homodimeric ancestors. The divergence of the sequence, structure, and target recognition behavior of homing endonucleases, as illustrated by this study, leads to the invasion of novel genomic sites by mobile introns during evolution. PMID- 16698549 TI - GTP-Ras disrupts the intramolecular complex of C1 and RA domains of Nore1. AB - The novel Ras effector mNore1, capable of inducing apoptosis, is a multidomain protein. It comprises a C1 domain homologous to PKC and an RA domain similar to the Ras effectors AF-6 and RalGDS. Here, we determine the affinity of these two domains to the active forms of Ras and Rap1 using isothermal calorimetric titration. The interaction of Ras/Rap1-GTP with the RA domain of mNore1 is weakened significantly by direct binding of the C1 domain to the RA domain. In order to analyze this observation in atomic detail, we solved the C1 solution structure by NMR. By determining chemical shifts and relaxation rates, we can show an intramolecular complex of C1-RA. GTP-Ras titration and binding to RA disrupts this complex and displaces the C1 domain. Once the C1 domain tumbles freely in solution, a lipid binding interface becomes accessible. Furthermore, we provide evidence of phosphatidylinositol 3-phosphate binding of the free C1 domain. PMID- 16698550 TI - Structures and polymorphic interactions of two heptad-repeat regions of the SARS virus S2 protein. AB - Entry of SARS coronavirus into its target cell requires large-scale structural transitions in the viral spike (S) glycoprotein in order to induce fusion of the virus and cell membranes. Here we describe the identification and crystal structures of four distinct alpha-helical domains derived from the highly conserved heptad-repeat (HR) regions of the S2 fusion subunit. The four domains are an antiparallel four-stranded coiled coil, a parallel trimeric coiled coil, a four-helix bundle, and a six-helix bundle that is likely the final fusogenic form of the protein. When considered together, the structural and thermodynamic features of the four domains suggest a possible mechanism whereby the HR regions, initially sequestered in the native S glycoprotein spike, are released and refold sequentially to promote membrane fusion. Our results provide a structural framework for understanding the control of membrane fusion and should guide efforts to intervene in the SARS coronavirus entry process. PMID- 16698551 TI - Structure of the mammalian NOS regulator dimethylarginine dimethylaminohydrolase: A basis for the design of specific inhibitors. AB - Dimethylarginine dimethylaminohydrolase (DDAH) is involved in the regulation of nitric oxide synthase (NOS) by metabolizing the free endogenous arginine derivatives N(omega)-methyl-L-arginine (MMA) and N(omega),N(omega)-dimethyl-L arginine (ADMA), which are competitive inhibitors of NOS. Here, we present high resolution crystal structures of DDAH isoform 1 (DDAH-1) isolated from bovine brain in complex with different inhibitors, including S-nitroso-L-homocysteine and Zn2+, a regulator of this mammalian enzyme. The structure of DDAH-1 consists of a propeller-like fold similar to other arginine-modifying enzymes and a flexible loop, which adopts different conformations and acts as a lid at the entrance of the active site. The orientation and interaction mode of inhibitors in the active site give insight into the regulation and the molecular mechanism of the enzyme. The presented structures provide a basis for the structure-based development of specific DDAH-1 inhibitors that might be useful in the therapeutic treatment of NOS dysfunction-related diseases. PMID- 16698552 TI - Two structurally atypical HEAT domains in the C-terminal portion of human eIF4G support binding to eIF4A and Mnk1. AB - The X-ray structure of the C-terminal region of human eukaryotic translation initiation factor 4G (eIF4G) has been determined at 2.2 A resolution, revealing two atypical HEAT-repeat domains. eIF4G recruits various translation factors and the 40S ribosomal subunit to the mRNA 5' end. In higher eukaryotes, the C terminus of eIF4G (4G/C) supports translational regulation by recruiting eIF4A, an RNA helicase, and Mnk1, the kinase responsible for phosphorylating eIF4E. Structure-guided surface mutagenesis and protein-protein interaction assays were used to identify binding sites for eIF4A and Mnk1 within the HEAT-repeats of 4G/C. p97/DAP5, a translational modulator homologous to eIF4G, lacks an eIF4A binding site in the corresponding region. The second atypical HEAT domain of the 4G/C binds Mnk1 using two conserved aromatic/acidic-box (AA-box) motifs. Within the first AA-box, the aromatic residues contribute to the hydrophobic core of the domain, while the acidic residues form a negatively charged surface feature suitable for electrostatic interactions with basic residues in Mnk1. PMID- 16698554 TI - Divergence of interdomain geometry in two-domain proteins. AB - For homologous protein chains composed of two domains, we have determined the extent to which they conserve (1) their interdomain geometry and (2) the molecular structure of the domain interface. This work was carried out on 128 unique two-domain architectures. Of the 128, we find 75 conserve their interdomain geometry and the structure of their domain interface; 5 conserve their interdomain geometry but not the structure of their interface; and 48 have variable geometries and divergent interface structure. We describe how different types of interface changes or the absence of an interface is responsible for these differences in geometry. Variable interdomain geometries can be found in homologous structures with high sequence identities (70%). PMID- 16698553 TI - Rounding up: Engineering 12-membered rings from the cyclic 11-mer TRAP. AB - The protein TRAP (trp RNA binding attenuation protein) forms a highly thermostable ring-shaped 11-mer. By linking in tandem two, three, or four DNA sequences encoding TRAP monomers, we have engineered new rings that consist of 12 TRAP subunits and bind 12 ligand molecules. The hydrogen bonding pattern and buried surface area within and between subunits are essentially identical between the 11-mer and 12-mer crystal structures. Why do the artificial proteins choose to make single 12-mer rings? The 12-mer rings are highly sterically strained by their peptide linkers and far from thermostable. That proteins choose to adopt a strained conformation of few subunits rather than an unstrained one with 11 subunits demonstrates the importance of entropic factors in controlling protein protein interactions in general. PMID- 16698555 TI - [Emergent viruses: SARS-associate coronavirus and H5N1 influenza virus]. AB - Two viral agents with RNA genome are responsible for emerging illnesses: influenza virus A/H5N1 and Severe Acute Respiratory Syndrome virus (SARS). For the diagnosis of SARS virus infection, an epidemiological investigation is necessary to know whether the patient has been exposed to a risk in a country where the SARS virus is circulating or whether the patient had worked in a laboratory handling SARS virus. The detection of SARS virus is possible in various clinical samples (including urine) by viral culture or RT-PCR. The handling of those samples and RNA extraction must be performed in a BSL3 laboratory. The SARS virus RT-PCR is poorly sensitive, therefore the test should be performed on samples collected consecutively for several days. In front of a suspicion of A/H5N1, similar procedures are recommended. An epidemiologic investigation is necessary to specify whether the patient stayed in a country where A/H5N1 virus was circulating. Clinical samples needed for a specific diagnosis are: nasopharyngeal, throat-swab or fecal samples, cerebrospinal fluid and blood. The presence of A/H5N1 virus is confirmed by viral isolation or RNA detection by RT-PCR. RNA extraction must be performed in a BSL3 laboratory. For diagnosis of A/H5N1 virus infection, RT-PCR test amplifies specifically a fragment of H5 gene (Hemagglutinin). In french laboratories of medical virology, procedures are ready to diagnose the first case of A/H5N1 virus infection and cases of reemerging SARS virus infection. PMID- 16698556 TI - [Heart fatty-acid binding protein (h-FABP): a new cardiac marker]. AB - Heart Fatty-Acid Binding Protein (h-FABP) is a small cytosolic protein that is abundant in the heart and found at lower concentrations in muscle or in the brain. h-FABP is released into the circulation shortly after the onset of ischemia. Several studies indicate its usefulness in cardiology: exclusion of acute myocardial infarction, detection of reperfusion, prognostic value... A rapid immuno-chromatographic assay (Cardiodetect) was recently commercialized in France with a result obtainable within 15 minutes. We review the strengths and weakness of h-FABP for detecting myocardial injury. PMID- 16698557 TI - [Parasitic etiology of blood hypereosinophilia]. AB - Blood hypereosinophilia is a common finding in medical practice requiring further investigations. Parasitic diseases are a major cause of eosinophilia and eosinophilia is mainly due to helminths. The investigation of eosinophilia is a long and complicated process. The etiologic diagnosis of eosinophilia requires a review of the patient's history which may reveal travel out of country, symptoms and rate and evolution of eosinophilia. For subjects originating from a country as Tunisia, suspected parasitosis -if eosinophilia is massif- are toxocariasis, distomiasis and scabies. Anguilluliasis and ankylostomiasis will be suspected if the subject is originating from some hot and wet focus. For subjects who have traveled to tropical country, bilharziasis, anguilluliasis and ankylostomiasis will be suspected. Since filariasis and onchocerciasis are diseases by accumulation, they will be suspected if the stay has been long. PMID- 16698558 TI - [Relation between lupus-antiphospholipids antibodies and heart disorders]. AB - BACKGROUND AND AIM: Antiphospholipids antibodies (APL) are autoantibodies found in lupus erythematosus and disorders like. Their frequency varies between 2 and 62% according to the literature. An increased frequency of cardiac disorders in antiphospholipids (APL) positive lupus has been reported. The aim of our study was to evaluate the role of APL as an independent risk factor of cardiac disorders in patients with systemic lupus erythematosus. MATERIAL AND METHOD: A prospective study during 14 months has been designed with the cooperation of dermatologic, internal medicine and cardiology departments of the Aristide Le Dantec hospital of Dakar. Platelets count (Beckmann Coulter analyzer), activated partial thromboplastin time (Diagnostiga stago analyzer) and antiphospholipids antibodies (Elisa) were determined. RESULTS: 37 patients affected by lupus were included in this study with a net feminine prevalence (89%); 8 (14.6%) had APL's significant results and 20 presented an echographic heart abnormality. The analysis of our data did not reveal an increased risk of cardiac diseases among APL positive lupic patients as compared to the negative group (p = 1). CONCLUSION: The presence of APL in patients with systemic lupus does not so seem to be an independant risk factor of heart diseases. PMID- 16698559 TI - [Dissemination of an extended spectrum beta-lactamase producing Klebsiella pneumoniae strains in a Tunisian university hospital center]. AB - On a period of four years (january 1999-december 2002), 49 strains (non redundant) of extended spectrum beta-lactamase-producing Klebsiella pneumoniae from 43 hospitalised patients and three strains from the environment hospital were collected at Mongi Slim University Hospital Center. The objectives of our work were to investigate for clonality of strains, to clarify transmission fashions and reservoirs of the infection by reviewing clinical records and typing strains. Antibiotic susceptibility testing, plasmid analysis and Random Amplified Polymorphic DNA (RAPD) have been done. 84% of the patients were hospitalized in the intensive care and pediatric units. Urinary infections and septicaemias were the more frequent infections (74.2%). The 49 isolats have been classified in 13 antibiotypes. The plasmid analysis showed 16 patterns. The RAPD revealed 28 patterns and variation within patterns in five cases. Our results showed a diversity of the strains suggesting endemicity, possible transmission of plasmids and persistence of some clones which circulated between services. The used markers permitted the evaluation of a long-term strategy of prevention requiring a strict observance of hygiene rules and rational use of antibiotics. PMID- 16698560 TI - [Prescribing guidance pocket chart on the initiation of warfarin therapy in a geriatric hospital: a useful but challenging tool]. AB - Elderly patients are at high risk of over-anticoagulation and of haemorrhagic risk when on warfarin, especially during treatment induction. In Charles Foix Hospital, a 800-bed geriatric hospital, we specifically developed for in-patients older than 70 years (target INR 2.5) a simple low-dose warfarin induction regimen. The dosing recommendations were summarized on a prescribing guidance pocket chart. Eighteen months after the distribution of the chart, we conducted a one-year observational study in order to evaluate: i/ the time needed to achieve the warfarin maintenance dose; ii/ the prescriber'adherence to the recommendations; iii/ the benefit for elderly patients receiving warfarin therapy. The mean time needed to achieve the warfarin maintenance dose was 12.3 +/- 7.0 days for the 89 patients included in the study: 10.6 +/- 5.9 days for the 30 patients whose prescribers followed the recommendations versus 13.5 +/- 7.6 days for the 59 patients whose prescribers did not follow the recommendations. There is a trend to a more frequent over-anticoagulation in patients whose prescribers did not follow the recommendations. The duration of the heparin warfarin overlap was significantly shorter when the recommendations were followed. Finally, the reasons for non-adherence to the recommendations were analyzed. This study illustrates an assessment of practice in an health care institution. PMID- 16698561 TI - [Primary hyperaldosteronism]. AB - Diagnosing primary aldosteronism, a hypertensive endocrine disease, is difficult because of an ill-defined frequency, various clinical forms and multiple diagnostic criteria. Current recommendations rest upon aldosterone and renin or renin activity determinations, the main point being to investigate aldosterone secretion with regards to of its main stimulus, renin. Proponents of the renin assay argue that it is easy and reliable. Proponents of the renin activity assay favour this method because of multiple epidemiological studies. Whatever the method used, each laboratory has to establish its critical thresholds in relation to the kits used. Extensive comparative studies would be useful to appreciate their relative benefits. PMID- 16698562 TI - [Evaluation of the automated coagulation analyser ACL TOP (instrumentation laboratory)]. AB - The ACL TOP is a fully automated random access analyser for coagulometric, chromogenic ou immunologic measurements. The aim of the present study was to evaluate its performances. The tests performed were: prothrombin time (PT), activated partial thromboplastin time (aPTT), prothrombin time derived fibrinogen (FibD), factor VIII (FVIII), antithrombin (AT) and free protein S antigen (PS). The comparison study was performed by analyzing patients samples in duplicate with ACL TOP and the analyser MDA II (BioMerieux) or CA 6000 (Dade-Behring). Imprecision (CV%) was < 4% for routine tests and < 5% for FVIII, AT and PS, except a between-run imprecision < 7,5% for low AT levels. The measured lower limits of linearity for FVIIII, AT and PS were satisfactory. The correlation observed between the ACL TOP and the other analysers was strict for PT and AT and good for TCA and FibD. Triglyceride levels > or = 6 mmol/L interfered with PT and FibD measurements. No sample or reagent carryover was observed in the conditions of the study. Overall, the ACL TO is an analyser with very satisfactory analytical and technical performances and is well suited for routine and specialized coagulation laboratories. PMID- 16698563 TI - [Diagnostic value of serum and pleural fluid adenosine deaminase activity in tuberculous pleurisy]. AB - The biological diagnosis of tuberculous pleurisy poses the problem of the time required to obtain results and of the sensitivity of the usual diagnostic methods. The determination of adenosine deaminase (ADA) activity has been proposed for the diagnosis of tuberculous pleural effusion and for the follow-up. In the present study, ADA in pleural effusion (p-ADA) and in serum (s-ADA) has been measured in 2 groups of patients: tuberculosis (27) and non-tuberculosis (53) patients. The upper limit of the normal values was fixed at 37 U/L. Comparing these 2 groups, we observed a specificity of 81.2% and a sensitivity of 66.6%. The PPV and the NPV were respectively 64.3% and 82.7%. We used p-ADA/s-ADA ratio for diagnosis of tuberculous pleural effusion, a threshold value of 1.8 gave a sensitivity of 82.6% and a specificity of 84.8%. Within the tuberculosis patient group, the activity of ADA decreased after the appropriate treatment initiation. Thus, the determination of ADA activity and/ or the p-ADA/s-ADA ratio, can help to recognize the tuberculosis origin of pleural effusions. PMID- 16698564 TI - [Fatal evolution of a dilated cardiomyopathy during a primary Epstein-Barr virus infection]. AB - We report the observation of a 2-year-old child who developed an acute heart failure during a primary EBV infection. The viral serological diagnosis showed CMV and EBV IgM positivity. The detection of EBV genome by PCR and the characteristic evolution of EBV serological response confirmed a primary EBV infection. PMID- 16698565 TI - [BNP/NT-proBNP: what is the best choice in an emergency laboratory?]. AB - BNP and NT-proBNP are both well established as diagnostic and prognostic markers for congestive heart failure (CHF). However it remains for the biologist to choose between these two biomarkers depending on his equipment availability. The aim of this study was to compare results obtained with the Biosite Triage BNP assay and the Dade Behring NT-proBNP assay with regards to the clinical status. One hundred twelve patients (average age 76 +/- 13 years) with acute dyspnea were including and stratified by diagnosis at presentation into 3 groups: patients without acute CHF (group I, n=50), patients with non-cardiac dyspnea and CHF history (group II, n=22) and patients with acute CHF (group III, n=40). Levels of both BNP and NT-proBNP were higher among patients with cardiac dyspnea (group III) than among patients with a non-cardiac dyspnea (BNP=740 pg/mL versus 84 pg/mL; p<0.001 / NT-proBNP=7.502 pg/mL versus 499 pg/mL; p<0.001). ROC analysis for BNP or NT-proBNP were not statistically different in patients with acute CHF (group III) compared with patients with a non-cardiac dyspnea (group I + II) (AUC=0.927 versus AUC=0.930, p=0.90). Neither there was a difference between ROC analysis for BNP or NT-proBNP in patients with cardiac dyspnea (group III) compared to patients with a non cardiac dyspnea (group I) (AUC=0.981 versus AUC=0.975, p=0.76). Measurement of BNP or NT-proBNP is of identical interest for the diagnosis of acute CHF in acute dyspnea. The BNP Biosite assay was faster because analysis is performed on whole blood. With regards to analytical performance, the NT-proBNP Dade Behring assay had a higher accuracy and is highly recommended for the follow-up of CHF treatment. PMID- 16698566 TI - [Human trichostrongyloidosis: case report and microscopic difficulties to identify ankylostomidae eggs]. AB - We present the case of a 53 year old woman living in a rural area and which suffered from gastric pain and high blood eosinophil polymorphonuclear counts for several years. The parasitological stool examination showed Trichostrongylus sp. larvae and eggs. Clinical and biological signs resolved after 2 courses of treatment with albendazole. We also report the microscopic difficulties to identify Ankylostomidae eggs. PMID- 16698567 TI - [Interest and limit of a free light chain immunoassay in serum and urine for the diagnosis and the follow-up of monoclonal dysglobulinemia]. AB - This work aims to define the interest and the limits of free light chain (FLC) determination in serum and urine for the investigation of monoclonal gammopathies. Based on the study of nine typical cases extracted from laboratory practice, the authors demonstrate the interest of this determination for the diagnosis and the monitoring of FLC and non secretory myelomas. This test is also useful for the evaluation of response to chemotherapy and the early detection of relapses in intact immunoglobulin multiple myelomas. These results are discussed in the light of the literature with a special emphasis on AL amyloidosis and monoclonal gammapathy of undetermined significance (MGUS). Finally the authors underline some limitations leading to an overestimation of the results in certain patients together with the difficulty to interpret data when a renal damage is associated. PMID- 16698568 TI - [Acquired Von Willebrand syndrome]. AB - We report a case of acquired von Willebrand syndrome in association with multiple myeloma, diagnosed in a 54 year-old man suffering from sudden onset of mucocutaneous bleeding. The acquired von Willebrand syndrome is a rare bleeding disorder with laboratory findings similar to those of inherited von Willebrand disease. Diagnosis, etiology and pathophysiology of acquired von Willebrand syndrome are reviewed to establish a differential diagnosis with inherited von Willebrand disease. Identification of the underlying disease responsible for the acquired von Willebrand factor defect and the bleeding diathesis is necessary to choose among the therapeutic options the appropriate treatment. PMID- 16698569 TI - Variations and reconstruction of the hepatic artery in liver transplantation. PMID- 16698570 TI - Modified techniques for adult-to-adult living donor liver transplantation. AB - BACKGROUND: Because of critical organ shortage, transplant professionals have utilized living donor liver transplantation (LDLT) in recent years. We summarized our experience in adult-to- adult LDLT with grafts of right liver lobe by a modified technique. METHODS: From January 2002 to August 2005, 24 adult patients underwent living donor liver transplantation with grafts of the right liver lobe at West China Hospital, Sichuan University, China. Twenty-two patients underwent modified procedures designed to improve the reconstruction of the right hepatic vein and the tributaries of the middle hepatic vein by interposing a great saphenous vein (GSV) graft and the anastomosis of the hepatic arteries and bile ducts. RESULTS: No severe complications and death occurred in all donors. In the first 2 patients, (patients 1 and 2), operative procedure was not modified. One patient suffered from "small-for-size syndrome" and the other died of sepsis with progressive deterioration of graft function. In the rest 22 patients (patients 3 to 24), however, the procedure of venous reconstruction was modified, and better results were obtained. Complications occurred in 7 recipients including acute rejection (2 patients), hepatic artery thrombosis (1), bile leakage (1), intestinal bleeding (1), left subphrenic abscess (1), and pulmonary infection (1). One patient with pulmonary infection died of multiple organ failure (MOF). The 22 patients underwent direct anastomosis of the right hepatic vein to the inferior vena cava (IVC), 9 direct anastomosis plus the reconstruction of the right inferior hepatic vein, and 10 direct anastomosis plus the reconstruction of the tributaries of the middle hepatic vein by interposing a GSV graft to provide sufficient venous outflow. Trifurcation of the portal vein was met in 3 patients. Venoplasty or separate anastomosis was performed. The ratio of graft to recipient body weight ranged from 0.72% to 1.17%. Among these patients, 19 had the ratio <1.0% and 4 <0.8%, and the ratio of graft weight to recipient standard liver volume was between 31.86% and 62.48%. Among these patients, 10 had the ratio <50% and 2 <40%. No "small-for-size syndrome" occurred in the 22 recipients who were subjected to modified procedures. CONCLUSIONS: With the modified surgical techniques for the reconstruction of the hepatic vein to obtain an adequate outflow and provide a sufficient functioning liver mass, living donor liver graft in adults using the right lobe can be safe to prevent the "small-for-size syndrome". PMID- 16698571 TI - Liver retransplantation: report of 80 cases and review of literature. AB - BACKGROUND: Because the orthotopic liver transplantation (OLT) was performed widely in recent 5 years throughout China, the proportion of recipients whose graft function deteriorated to be retransplantation candidates increased gradually. This study was undertaken to analyze clinical experience of orthotopic liver retransplantation (re-OLT) at our center. METHODS: The medical records of 80 patients who had undergone liver retransplantation at our center from January 1999 to July 2005 were analyzed retrospectively, including indications and timing of retransplantation, surgical techniques, and the causes of death. RESULTS: The commonest cause leading to hepatic graft loss and subsequent retransplantation was biliary complications in 36 patients (45%). The patients underwent retransplantation more than 30 days after their primary transplant recovered better than those who underwent retransplantation within 8-30 days after primary transplantation (perioperative mortality 19.6% versus 70%). Sepsis (12 of 22 patients, 54.5%) and multiple organ failure (4 of 22 patients, 18.2%) were leading causes of re-OLT recipient deaths. CONCLUSIONS: Proper indications and optimal operative time, surgical procedures, perioperative monitoring and appropriate postoperative treatment contribute to the improvement of the survival rate of patients after liver retransplantation. PMID- 16698572 TI - Cardiac arrest after graft reperfusion during liver transplantation. AB - BACKGROUND: Cardiac arrest (CA) during orthotopic liver transplantation (OLT) is rare but it threatens the lives of patients. The cause of perioperitive CA is not fully understood. We reported the occurrence of CA in 5 patients after unclamping of the vena cava and investigated the relationship between CA and associated variables. METHODS: Five patients with CA after graft reperfusion during OLT in our unit from November 1996 to September 2003 were retrospectively reviewed. Analyzed data included donor and recipient demographic data, and recipient operative and postoperative events. RESULTS: Five (2.1%) of 240 patients undergoing OLT experienced CA 5 minutes after graft reperfusion. Two patients died of resuscitation failure. Hyperkalemia and metabolic acidosis after revascularization were observed in some patients. The five patients had hypothermia and hypocalcemia, and one had pulmonary embolism. CONCLUSIONS: CA is one of the syndromes after reperfusion. Many factors such as hyperkalemia, acidosis or pulmonary embolism combined with hypothermia and hypocalcemia during the operation seem to contribute to the occurrence of CA. PMID- 16698573 TI - Drug resistance of infectious pathogens after liver transplantation. AB - BACKGROUND: Infection in liver recipients is related to high risk of transplantation failure and mortality. Infectious agents isolated from 55 liver recipients from January 2003 through June 2005 were studied to improve the anti infectious therapy. METHODS: Pathogens were isolated from routine culture. K-B method was used to examine the drug susceptibility. Extended spectrum beta lactamase, AmpC beta-lactamase and Van gene in E.coli were examined by the agar dilution susceptibility test and Nitrocefin test. RESULTS: Thirty-nine of the 55 recipients got infection. The 513 strains of pathogens isolated from 1861 specimens were predominantly Gram negative bacteria and over 40% of them showed resistance to more than 4 drugs. The positive rates of extended spectrum beta lactamse and AmpC beta-lactamse production in E.cloacae were 32.4% and 36.8%, in E.coli were 33.8% and 10.5%, but the rates of these 2 bacteria producing both lactamses were 24.3% and 7.0%. The beta-lactamse production rates of Enterococcus faecalis and Enterococcus faecium were 8.8% and 11.1%, and the resistance rates to vancomycin were 11.2% and 18.5%, respectively. CONCLUSIONS: Infectious pathogens isolated from liver recipients are potent and multiple drug resistant. ESBLs and AmpC beta-lactamases are the major factors associated with Gram negative drug resistance. The infection of Enterococcal species presents as a particular challenge. PMID- 16698574 TI - Influence of perfusion by gaseous oxygen persufflation on rat donor liver. AB - BACKGROUND: Hepatic ischemia-reperfusion injury is one of the major problems in liver transplantation. This study aimed to investigate the possible improvement of aerobic metabolism of ischemic donor livers through systemic persufflation with gaseous oxygen in rinse solution. METHODS: Thirty rats were randomly divided into 3 groups. In group A (control, n=10), the livers were perfused with 4 degree centigrade lactic acid ringer's solution through the cannulated portal vein for 25 minutes soon after warm ischemia for 30 minutes. In group B (n=10), the livers were treated the same way as in group A except for addition of gaseous oxygen in the rinse solution. In group C (n=10), the livers were treated similarly as in group B except for addition of superoxide dismutase (SOD) in the rinse solution. RESULTS: In group B, the malondialdehyde (MDA)levels in hepatic tissues after perfusion were significantly increased (P<0.01), whereas the SOD levels were markedly decreased (P<0.01). After combined use with antioxidant in group C, the levels of MDA and SOD in hepatic tissues after perfusion recovered significantly (P<0.01). CONCLUSION: Perfusion by gaseous oxygen persufflation in combination with antioxidative agents is promising for resuscitating the ischemically altered livers for successful transplantation. PMID- 16698575 TI - Enteral feeding of glycyl-glutamine dipeptide improves the structure and absorptive function of the small intestine after allogenetic liver transplantation in rats. AB - BACKGROUND: Recipients of liver transplantation could have postoperative structural injury and declined absorptive function in the gastrointestinal tract. Glutamine (Gln) is a special nutrient of small intestinal mucosa and of various kinds of cells proliferating rapidly. But Gln could form a kind of poisonous pyroglutamic acid in water solution, which is the limitation of Gln in clinical practice. Glycyl-glutamine (Gly-Gln) is highly soluble and can be hydrolyzed to release glutamine. This study was undertaken to observe the effect of Gly-Gln dipeptide by enteral feeding on the intestinal structure and absorptive function after allogenetic liver transplantation in rats. METHODS: Twelve male inbred Lewis rats were selected randomly as donors, and 24 male inbred BN rats as recipients of allogenetic liver transplantation. The recipients were also randomly divided into two groups: control group (ALA group, n=12) and experimental group (GLN group, n=12). In each group, 6 normal BN rats were sampled as the normal parameter on the 3rd preoperative day. The 6 recipients in the control group received alanine 0.6 g/kg daily for 3 days before operation and 7 days after operation by gastric perfusion, and the 6 recipients in the experimental group were given Gly-Gln 0.6 g/kg daily the same way. The 12 BN recipients underwent 3-day fasting (free access to water with 0.23% sodium chloride) and orthotopic liver transplantation in aseptic conditions and were given subcutaneous injection of CsA 2 mg/kg daily after the operation. The 12 BN recipients were sampled on the 8th postoperative day. All of the 24 BN rats were subjected to examination of mucosal structure, activities of Na+-K+-ATP and disaccharidase, and D-xylose absorption test. RESULTS: The 12 BN recipients were alive after liver transplantation. On the 3rd preoperative day, mucosal structure, activities of Na+-K+-ATP and disaccharidase and D-xylose absorption in the two groups were not significantly different. On the 8th postoperative day, the parameters of the two groups were markedly changed compared with those on the 3rd preoperative day. However, the parameters of GLN group were remarkably higher than those of ALA group. CONCLUSION: Enteral feeding of Gly-Gln could improve the structure and absorptive function of the small intestine after liver transplantation in rats. PMID- 16698576 TI - Effect of Shenfu injection on ischemia-reperfusion injury of rat liver graft. AB - BACKGROUND: It is reported that Shenfu injection (an injection prepared from traditional Chinese medicines red ginseng and aconite root) can decrease the extent of ischemia-reperfusion injury to many organs, such as the heart and kidney. We therefore investigated the effect of Shenfu injection on ischemia reperfusion injury of rat liver graft and its mechanism. METHODS: Male Sprague Dawley (SD) rats were used as a model for isogeneic orthotopic liver transplantation. Sixty rats were randomly divided into two groups (30 in each group). The recipient was given intravenous Shenfu injection immediately before the removal of the liver in the Shenfu group and normal saline of the same volume in the control group. At 3, 6 and 24 hours after the reperfusion, blood and hepatic tissue were taken for examination. RESULTS: The levels of superoxide dismutase (SOD) and nitric oxide (NO) increased more significantly in the Shenfu group than in the control group (P<0.05). The levels of serum liver enzymes, hyaluronic acid (HA), malondialdehyde (MDA), tumor necrosis factor-alpha (TNF alpha), interleukin-1 (IL-1), endothelin -1 (ET-1) and liver cell apoptosis index were lower in the Shenfu group than in the control group (P<0.05). Microscopic examination revealed that the morphological changes of hepatic tissue were more severe in the control group than in the Shenfu group. CONCLUSIONS: Shenfu injection has protective effect on ischemia-reperfusion injury of rat liver graft. It inhibits the production of oxygen free radical and the activation of Kupffer cells, decreases apoptosis of liver cell, and improves microcirculation. PMID- 16698577 TI - Oxidative stress in humans following the Pringle manoeuvre. AB - BACKGROUND: Oxidative stress is induced in the liver by application of the Pringle manoeuvre. Malondialdehyde is a carbonyl compound formed during lipid peroxidation and prostaglandin biosynthesis, which combines with DNA to form a number of adducts. Among them is the DNA adduct; 3-(2-deoxybeta dierythropentafuranosyl) pyr [1, 2-alpha]-purin-10 (3H) one or M1G. This study was undertaken to determine the suitability of M1G as a novel marker of ischemia reperfusion injury in the liver and its correlation with both the length of Pringle clamp application and the overall length of the operation. METHODS: Normal and colorectal liver metastatic tissues were obtained in 12 patients before and after application of the Pringle manoeuvre. All samples were snap frozen in liquid nitrogen at -80 degree centigrade. DNA was extracted and M1G quantification was performed by immunoslotblot analysis. RESULTS: M1G levels in normal liver tissue were 4.0+/-1.0 per 10(7) nucleotides before the Pringle manoeuvre and 7.4 +/-1.0 per 10(7) nucleotides after the Pringle manoeuvre (mean+/-standard deviation) (P<0.05 by ANOVA). M1G levels in malignant liver tissue were 2.5+/-1.4 per 10(7) nucleotides before the Pringle manoeuvre and 6.5+/-1.9 adducts per 10(7) nucleotides after the Pringle manoeuvre (P<0.05). Adduct levels in normal liver tissue showed a significant correlation with cumulative period of Pringle application. CONCLUSIONS: This is the first time that the tissue levels of M1G before and after application of the Pringle manoeuvre have been studied. The results show that the Pringle manoeuvre exerts significant oxidative stress in human hepatocytes, which is Pringle-time dependent. The results highlight the potential for oxidative DNA adducts levels as a tool for measuring the severity of ischemia-reperfusion injury. PMID- 16698578 TI - Effects of hemoglobin concentration on hyperdynamic circulation associated with portal hypertension. AB - BACKGROUND: Chronic anemia caused by different factors is a common complication in patients with portal hypertension, but attention has been rarely paid to its detrimental effect on hemodynamic status. This study was undertaken to investigate the effects of hemoglobin concentration on hyperdynamic disturbance associated with portal hypertension (PHT). METHODS: According to blood hemoglobin level of 120 g/L, 55 patients with portal hypertension were divided into two groups, anemic and nonanemic. Hemodynamic and clinical data of the patients were analyzed retrospectively. The data were analyzed separately according to the Child classification in an attempt to avoid the effects of differences on hepatic function. RESULTS: Compared with the nonanemic group, the anemic group had an increased cardiac output (7.4+/-1.7 L/min vs. 6.3+/-1.9 L/min, P=0.028), free portal pressure (29.1+/-3.1 mmHg vs. 26.8+/-3.3 mmHg, P=0.012), a decreased mean arterial pressure (84.0+/-10.7 mmHg vs. 97.1+/-12.0 mmHg, P<0.01), and systemic vascular resistance (866+/-215 dyn/s.cm5 vs. 1207+/-317 dyn/s.cm5, P<0.01). Similar results were obtained when Child A and Child B-C patients were analyzed separately. Multivariate logistic regression revealed that the concentrations of hemoglobin (standard coefficient=0.31, P=0.01) and albumin (standard coefficient=0.21, P=0.04) were independent factors influencing the systemic vascular resistance in PHT patients. CONCLUSIONS: Anemia aggravates the hyperdynamic circulation of portal hypertension. Hemoglobin concentration is an important variable for evaluating the degree of hemodynamic disturbance in PHT patients. PMID- 16698579 TI - Relationship between CT grouping and complications of liver cirrhosis. AB - BACKGROUND: Imaging examination is important for hepatic cirrhosis. But the relationship between magnetic resonance (MR), computed tomography (CT) or ultrasound findings and pathological groups, degree, or reserve function of the cirrhotic liver is not clear. In this study, we investigated the relationship between the CT groupings of liver cirrhosis and its complications and clinical conditions. METHODS: The CT findings in 357 patients with liver cirrhosis were grouped. The complications were analyzed, included splenomegaly, varicose collateral veins, ascites, pleurorrhea, primary liver carcinoma, gallbladder stone, etc. Blood routine (BRt), and serum usea nitrogen (SUN), creatinine and uric acid were measured and hypersplenia and liver-kidney syndrome were estimated. RESULTS: Three hundred and fifty-seven patients with cirrhosis were divided into homogeneous group (87 patients, 24.4%), segmental group (41, 11.5%), and nodal group (229, 64.2%). The grade of spleen enlargement in the segmental and the nodal groups was significantly greater than that in the homogeneous group (P<0.01 and P<0.001). The patients with varices were shown in a descending order in the segmental group (70.7%), the nodal group (17.0%) and the homogeneous group (2.3%), respectively. Significant difference was observed among the 3 groups (P<0.001). Ascites was seen in 182 patients (79.5%) of the nodal group, in 11 patients (26.8%) of the segmental group and in 9 patients (10.3%) of the homogeneous group (P<0.001). Sixty-eight patients (29.7%) in the nodal group had primary liver carcinoma and 1 (2.4%) in the segmental group and 5 (5.8%) in the homogeneous group (P<0.001). The number of patients with decreased concentration of hematoglobin in the nodal group was more than that in the homogeneous group (P<0.001). The mean values of hematoglobin and platelet in the nodal group and the segmental group were significantly lower than those in the homogeneous group (P<0.05). The number of patients with increased concentration of SUN in the nodal group and the segmental group was more than that in the homogeneous group (P<0.005). The concentration of SUN in the nodal group was significantly higher than that in the homogeneous group (P<0.002). CONCLUSION: There is a close relationship between the grouping of liver cirrhosis by CT findings and complications caused by the cirrhosis. The grouping is significant for estimating clinical conditions. PMID- 16698580 TI - Adjuvant hepatic chemotherapy after resection of solitary hepatocellular carcinoma associated with hepatitis B virus cirrhosis. AB - BACKGROUND: Although resection is the major treatment for patients with hepatocellular carcinoma (HCC), the high intrahepatic recurrence remains a cardinal cause of death. This study was undertaken to evaluate the effect of hepatic arterial infusion chemotherapy on the survival and recurrence of HCC patients with hepatitis B virus (HBV) cirrhosis after resection. METHODS: Twenty eight patients who had undergone placement of a hepatic arterial pump at the time of liver wedge resection for HCC from 1998 through 2004 were reviewed retrospectively. These patients aged 23-71 years had HBV cirrhosis (Child-Pugh class A or B). They were given floxuridine (FUDR)(250 mg), doxorubicin (10 mg) and mitomycin C (4 mg) alternatively every 2 or 3 days through arterial pumps for 8 cycles each year in the first two years after resection. Meanwhile, traditional Chinese herbal medicine was prescribed to the patients. When the leukocyte count was as low as 3X10(9)/L or asparate aminotransferase (AST) level was significantly increased, the regimen of chemotherapy was delayed for the normalization of leukocyte count and AST level (below 80 U/L). RESULTS: Of the 28 patients, 23 received 8 or 16 cycles of the set regimen of chemotherapy. These patients are alive with no evidence of recurrence. Among them, 5, 7, and 11 patients are alive beyond 5 years, 3 years, and 1 year respectively. In the remaining 5 patients, 3 who had had a HCC 10 cm or more in diameter showed tumor recurrence within 1 year, in whom, 8 cycles of chemotherapy were not completed because of their low leukocyte count (<3 x 10(9)/L) and poor liver function. One patient who had received 8 cycles of chemotherapy demonstrated recurrence at 16 months after resection. One patient who had received 16 cycles of chemotherapy had intrahepatic recurrence at 58 months after surgery. No recurrence was observed in 17 patients who had received 16 cycles of chemotherapy. CONCLUSION: Adjuvant hepatic arterial chemotherapy may be feasible to improve the survival of patients after resection of solitary HCC associated with HBV cirrhosis. PMID- 16698581 TI - Bacteriological and electron microscopic examination of primary intrahepatic stones. AB - BACKGROUND: Primary intrahepatic cholelithiasis is usually combined with biliary tract infection. This research was undertaken to investigate the relationship between intrahepatic stones and biliary tract infection. METHODS: Thirty-five bile samples and 30 stones specimens were cultured for bacteria and 12 stones specimens were examined with a scan electron microscope (SEM) or a transmission electron microscope (TEM). RESULT: 94.2% bile samples and 96.7% stones specimens were positive in bacteria culture. Bacteria were found in stones under SEM and TEM. CONCLUSION: Bacteria in stones are associated with the infection of the biliary tract. PMID- 16698582 TI - Relationship between pre-TIPS hepatic hemodynamics and postoperative incidence of hepatic encephalopathy. AB - BACKGROUND: Hepatic encephalopathy (HE) is one of the complications that have limited the effectiveness of transjugular intrahepatic portosystemic shunt (TIPS) most significantly. Up to the present, the predicting factors of HE post-TIPS have been debated controversially. This study was undertaken to verify the relationship between pre-TIPS intrahepatic hemodynamics and the incidence of post TIPS HE. METHODS: The hepatic blood dynamics was evaluated in 41 patients with liver cirrhosis before TIPS and at one month after TIPS by ultrasonography. The patients were divided into two groups according to Doppler findings before TIPS: group 1, patients with prograde portal flow, and group 2, patients with hepatofugal or back-forth portal flow. The clinical characteristics (age, sex, etiology of liver disease, pre-TIPS Child-Pugh score, incidence of pre-TIPS HE, and portacaval pressure gradient), incidence of post TIPS HE, and pre-/post-TIPS hepatic arterial resistant index (RI) in the two groups were compared. The independent prognostic value of pre-TIPS variables for the onset of HE after TIPS, including age, Child-Pugh score, presence of HE before TIPS, and the pattern of portal flow, was tested with a multiple-factor regression analysis. RESULTS: No significant difference in age, etiology of liver disease, indications of TIPS placement, incidence of HE before TIPS, and portacaval gradient before and after TIPS was observed between the two groups; but liver failure was more severe in group 2 (P<0.05). The incidence of post-TIPS HE in group 2 was significantly lower than that in group 1 (P<0.01). Pre-TIPS, the RI of the hepatic artery in group 1 was significantly higher than that in group 2 (P<0.01). However, TIPS induced a significantly decreased RI in group 1 (P<0.01), but not in group 2. Multiple-factor regression analysis demonstrated that the pattern of portal flow before TIPS was closely associated with the onset of post TIPS HE. CONCLUSIONS: Pre-TIPS intrahepatic hemodynamics is closely related to the incidence of post-TIPS HE. Hepatic hemodynamics of patients with hepatofugal portal blood flow only changes a little after TIPS and still provides compensatory blood supply of the hepatic artery, and the hepatic function is less affected. Hence HE is unlikely. Hepatic hemodynamics of patients with prograde portal blood flow changes a lot after TIPS, and dual blood supply of the portal vein and hepatic artery changes into compensatory blood supply of the hepatic artery, and hepatic function suffers greatly in a short time. Thus HE is mostly likely. PMID- 16698583 TI - Screening and evaluation of human single-chain fragment variable antibody against hepatitis B virus surface antigen. AB - BACKGROUND: Phage display technology has become a vital tool in studies aimed at identifying molecules binding to a specific target. It enables the rapid generation and selection of high affinity, fully human antibody product candidates to essentially any disease target appropriate for antibody therapy. In this study, we prepared the recombinant single-chain fragment variable (ScFv) antibody to hepatitis B virus surface antigen (HBsAg) by the phage display technology for obtaining a virus-targeting mediator. METHODS: mRNA was isolated from B-lymphocytes from a healthy volunteer and converted into cDNA. The fragment variables of heavy and light chain were amplified separately and assembled into ScFv DNA with a specially constructed DNA linker by polymerase chain reaction. The ScFv DNA was ligated into the phagmid vector pCANTAB5E and the ligated sample was transformed into competent E.coli TG1. The transformed cells were infected with M13K07 helper phage to form a human recombinant phage antibody library. The volume and recombinant rate of the library were evaluated by bacterial colony count and restriction analysis. After two rounds of panning with HBsAg, the phage clones displaying ScFv of the antibody were selected by enzyme-linked immunosorbant assay (ELISA) from the enriched phage clones. The antigen binding affinity of the positive clone was detected by competition ELISA. HB2151 E.coli was transfected with the positive phage clone demonstrated by competition ELISA for production of a soluble form of the anti-HBsAg ScFv. ELISA assay was used to detect the antigen binding affinity of the soluble anti-HBsAg ScFv. Finally, the relative molecular mass of soluble anti-HBsAg ScFv was measured by SDS-PAGE. RESULTS: The variable heavy (VH) and variable light (VL) and ScFv DNAs were about 340 bp, 320 bp and 750 bp, respectively. The volume of the library was up to 2 x 10(6) and 8 of 10 random clones were recombinants. Two phage clones could strongly compete with the original HBsAb for binding to HBsAg. Within 2 strong positive phage clones, the soluble anti-HBsAg ScFv from one clone was found to have the binding activity with HBsAg. SDS-PAGE showed that the relative molecular weight of soluble anti-HBsAg ScFv was 32 kDa. CONCLUSION: The anti-HBsAg ScFv successfully produced by phage antibody technology may be useful for broadening the scope of application of the antibody. PMID- 16698584 TI - Down-regulation of the gax gene in smooth muscle cells of the splenic vein of portal hypertension patients. AB - BACKGROUND: The expression of gax, an anti-proliferative homeobox gene, is rapidly down-regulated in vascular smooth muscle cells (VSMCs) following arterial injury. Whether the down-regulation of gax is involved in modulating the proliferation of smooth muscle cells of the splenic vein in patients with portal hypertension has not yet been elucidated. The aim of this study was to investigate the expression of the mRNA of the gax gene in smooth muscle cells of the splenic vein in patients with portal hypertension. METHODS: Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the expression of gax mRNA and immunohistochemistry staining was performed to demonstrate the expression of PCNA protein in the splenic veins of 28 patients with portal hypertension and those of 12 normal controls. This study was approved by the Institutional Ethics Committee and informed consent was obtained from all participants. RESULTS: RT-PCR showed that the expression of gax mRNA was PCNA positive and negative in the splenic vein of patients with portal hypertension (1.08+/-0.04 and 1.39+/-0.27, respectively). There was a significant difference in the 28 patients compared with the 12 normal controls (P<0.01). The relative expression of PCNA protein in the vascular tissues was significantly higher in the experimental group than in the control group. CONCLUSIONS: Down-regulation of gax mRNA and the overexpression of PCNA protein were seen in smooth muscle cells of the splenic vein in patients with portal hypertension, regulating the proliferation, migration and phenotypic change and resulting in remodelling of the splenic vein, which may play an important role in the pathogenesis and maintenance of portal hypertension. PMID- 16698585 TI - Negatively regulating mechanism of Sirpalpha1 in hepatocellular carcinoma: an experimental study. AB - BACKGROUND: Signal regulatory protein (Sirp) is a recently isolated, cloned and identified inhibitor receptor distributed in the membrane of hematopoietic and nonhematopoietic cells. Sirp alpha1 (Sirpalpha1) is a member of Sirp families. Sirpalpha1 can bind SHP-2 in the form of tyrosine phosphorylation by SH2 effect and negatively regulate growth factor, oncogene, or insulin-induced responses as its substrate. This study aimed to preliminarily clarify the negatively regulating proliferation mechanism of Sirpalpha1 in liver cancer. METHODS: pLXSN, Sirpalpha1 and Sirpalpha1P4Y2 plasmids were respectively transfected into Sk-Hep1 liver cancer cell line, and various stable Sk-Hep1 cell lines were obtained with screening agent of G418 (1200 microg/ml). The expressing levels of cyclin D1, CDK4, Fas, beta-catenin and gankyrin in various cell lines were determined with Western blotting. Cell cycles were determined at 0, 12 and 24 hours with flow cytometry after various synchronous cell lines were cultured without serum for 72. Cell apoptosis induced with agent of TNF-alpha (50 ng/ml) was determined with flow cytometry at 0, 0.5, 1, 3, 6 and 12 hours. RESULTS: Sirpalpha1 could significantly decrease the expression of cyclin D1, beta-catenin and gankyrin, but it couldn't affect the expression level of CDK4 and Fas. When synchronous cells were cultured for 12 hours, S phase Sk-Hep1 cell transfected with Sirpalpha1 plasmid was the lowest [(31.92+/-0.22)% vs. other cell lines, P<0.05], and the cell line was highly sensitive to TNF-alpha agent for 1 hour. (59.31+/ 0.59)% of apoptotic cells occurred (vs. the other time points, P<0.05). CONCLUSIONS: Sirpalpha1 might block the cell cycle of liver cancer, inhibit cell proliferation, promote cell apoptosis by decreasing the expression of cyclin D1, beta-catenin and gankyrin. It is one of the important mechanisms inhibiting the occurrence and development of hepatocellular carcinoma. PMID- 16698586 TI - Expression of cyclooxygenase in hyperdynamic portal hypertensive rats. AB - BACKGROUND: By detecting hemodynamic changes, concentration of plasm prostacyclin (PGI2) and expression of cyclooxygenase (COX) in vasculature and splanchnic tissues, we evaluated the relative contributions of PGI2 and COX mRNA expression to the hyperdynamic circulatory state in chronic portal hypertensive rats. METHODS: Fifty male Sprague-Dawley rats were divided into 3 groups: intrahepatic portal hypertension (IHPH, n=18) by injection of CCl4, prehepatic portal hypertension (PHPH, n=18) by partial stenosis of the portal vein, and sham operated controls (SO, n=14). Splanchnic hemodynamics was measured by radioactive microsphere techniques and the concentration of PGI2 was detected by specific radioimmunoassay for its stable hydrolysis product 6-keto-PGF1alpha. Semi quantitive reverse transcriptase-polymerase chain reaction (RT-PCR) was performed to measure the levels of COX-1 mRNA and COX-2 mRNA in the thoracic aorta, superior mesenteric artery (SMA), and small intestine of IHPH, PHPH and SO rats, respectively. RESULTS: Hyperdynamic circulatory state was characterized by increased splanchnic blood flow and decreased splanchnic vascular resistance in IHPH and PHPH rats. The concentration of plasma 6-keto-PGF1alpha (pg/ml) in IHPH (1093.75+/-142.15) and PHPH (897.42+/-53.29) rats was significantly higher than that in SO rats (730.13+/-98.67) (P<0.05). The expression of COX-1 mRNA in the thoracic aorta, SMA and small intestine was enhanced, whereas COX-2 mRNA expression was not detected in either of these vessels or the small intestine. The plasma 6-keto-PGF1alpha concentration and the expression of COX-1 mRNA in these vessels and the small intestine were closely correlated with such hemodynamic parameters as portal venous inflow (PVI), splanchnic vascular resistance (SVR) and free portal venous pressure (FPP) (P<0.05). CONCLUSION: The expression of COX-1 mRNA and the levels of PGI2 were closely related to the hyperdynamic circulatory state of portal hypertensive rats. PMID- 16698587 TI - Gradual upregulation of OCI-5 expression during occurrence and progression of rat hepatocellular carcinoma. AB - BACKGROUND: OCI-5, the rat homologene of human glypican 3 (GPC3), is confirmed upregulated in hepatocellular carcinoma (HCC). The present study was undertaken to detect gene expression change of OCI-5 during occurrence and progression of rat HCC. METHODS: Male Sprague-Dawley rats were given diethylnitrosamine (DENA) to induce HCC. Three DENA-induced rats and one control rat were sacrificed every week for 18 weeks during the development of HCC. Tissues specimens were snap frozen in liquid nitrogen and total RNA was isolated. Sk-Hep1 cells were treated with DENA at different concentrations. The gene expression levels of OCI-5 and GPC3 were detected with the RT-PCR method. RESULTS: OCI-5 was not expressed in normal rat liver tissues. When HCC occurred and aggravated, OCI-5 expression was gradually elevated to a very high level. GPC3 was not expressed in the DENA treated Sk-Hep1 cells. CONCLUSIONS: OCI-5 was not expressed in normal rat liver tissues but in rat HCC tissues. High-expression of OCI-5 in DENA-induced rat HCC model was the gene expression change of HCC not the DENA-induced gene expression. The expression level of OCI-5 was not only elevated in rat HCC but also gradually along the occurrence and progression of HCC, indicating that GPC3 might serve as a sensitive marker of early stage HCC. PMID- 16698588 TI - Tea polyphenols inhibit expressions of iNOS and TNF-alpha and prevent lipopolysaccharide-induced liver injury in rats. AB - BACKGROUND: Tea polyphenols have been shown to protect against carbon tetrachloride (CCl4)-induced liver injury, liver fibrosis, hepatic ischemia reperfusion injury. In this study, we examined the effect of tea polyphenols on lipopolysaccharide (LPS)-induced liver injury, and explored its mechanisms. METHODS: Sprague-Dawley rats received tea polyphenols (100 mg.kg-1.d-1) or vehicle (water) intragastrically by gavage for 14 days, followed by LPS (5 mg/kg) or saline injection intraperitoneally. Liver injury was assessed by biochemical assay and pathological analysis. Serum tumor necrosis factor-alpha (TNF-alpha) levels and liver malondialdehyde (MDA) contents were determined. Inducible nitric oxide synthase (iNOS) protein and TNF-alpha, iNOS and endothelial nitric oxide synthase (eNOS) mRNA expressions in the liver were detected by immunohistochemistry and reverse transcriptase-polymerase chain reaction (RT PCR), respectively. RESULTS: Administration of LPS resulted in liver injury in rats, evidenced by elevated activities of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), hepatocellular necrosis, and neutrophil infiltration in the liver. These responses were associated with increased serum TNF-alpha levels, induced iNOS protein, expressions of TNF-alpha, iNOS mRNA in the liver and elevated lipid peroxidation at 90 minutes or 6 hours after LPS injection. Pretreatment with tea polyphenols attenuated LPS-induced liver injury, and blunted the rises of serum TNF-alpha levels and lipid peroxidation and the induction of expressions of TNF-alpha, iNOS in the liver. CONCLUSION: Tea polyphenols prevent LPS-induced liver injury, and the mechanisms may involve the reduction of serum TNF-alpha levels and lipid peroxidation and the suppression of TNF-alpha, iNOS expressions in the liver. PMID- 16698589 TI - Effect of tea polyphenol on cytokine gene expression in rats with alcoholic liver disease. AB - BACKGROUND: Oxidative stress plays a pathogenetic role in initiation and progression of hepatic damage caused by alcohol. Recently, antioxidants have received considerable attention. Green tea polyphenols have both antioxidant and antiinflammatory properties. This study was designed to evaluate the effect of tea polyphenol (TP) on alcohol-induced liver injury in rats. METHODS: The rats were divided randomly into 3 groups: group A gastrically infused with alcohol for 12 weeks, group B fed with alcohol plus TP (250 mg/kg.d) simultaneously, and group C (control group) gastrically infused with normal saline. At the end of 12 weeks, the rats were sacrificed. The liver specimen of each rat was taken for histological examination. All data were statistically analyzed in quantum and semi-quantum. Gene expression of cytokines of each group was determined. RESULTS: At the end of 12 weeks, hepatic injury of different degrees developed in group A and group B compared to group C. The degree of hepatic injury was attenuated in group B, with slight steatosis, liver cellular swelling in small areas; less spot and focal necrosis, no bridging necrosis, less mega-bubble steatosis and less collagen deposition in contrast to group A. Gene expressions of IL-3, IL-4, IL 1R2, IL-6R, IL-7R2 were up-regulated in group B compared with group A, but those of IL-3Ra, IL-1R1 were down-regulated. Gene expressions of IL-13, IL-1R1, IL-7R2, EPO-R, LIFR were up-regulated in group A compared with group C, but those of IL 1R2, IL-5R2, CSF1, CD27, IL-6R were down-regulated. CONCLUSION: TP is able to attenuate alcoholic liver damage. Cytokine may contribute to alcoholic liver disease. PMID- 16698590 TI - Endoscopic management of postoperative bile leaks. AB - BACKGROUND: Significant bile leak as an uncommon complication after biliary tract surgery may constitute a serious and difficult management problem. Surgical management of biliary fistulae is associated with high morbidity and mortality. Biliary endoscopic procedures have become the treatment of choice for management of biliary fistulae. METHODS: Ninety patients presented with bile leaks after cholecystectomy (open cholecystectomy in 45 patients, cholecystectomy with common bile duct exploration in 20 and laparoscopic cholecystectomy in 25). The presence of bile leaks was confirmed by ERCP and the appearance of bile in percutaneous drainage of abdominal collections. Of the 90 patients with postoperative bile leaks, 18 patients had complete transaction of the common bile duct by ERCP and were subjected to bilioenteric anastomosis. In the remaining patients after cholangiography and localization of the site of bile leaks, therapeutic procedures like sphincterotomy, biliary stenting and nasobiliary drainage (NBD) were performed. If residual stones were seen in the common bile duct, sphincterotomy was followed by stone extraction using dormia basket. Nasobiliary drain or stents of 7F size were placed according to the standard techniques. The NBD was removed when bile leak stopped and closure of the fistula confirmed cholangiographically. The stents were removed after an interval of 6-8 weeks. RESULTS: Bile leaks in 72 patients occurred in the cystic duct (38 patients), the common bile duct (30), and the right hepatic duct (4). Of the 72 patients with post-operative bile leak, 24 had associated retained common bile duct stones and 1 had ascaris in common bile duct. All the 72 patients were subjected to therapeutic procedures including sphincterotomy with stone extraction followed by biliary stenting (24 patients), removal of ascaris and biliary stenting (1), sphincterotomy with biliary stenting (18), sphincterotomy with NBD (12), biliary stenting alone (12), and NBD alone (5). Bile leaks stopped in all patients at a median interval of 3 days (range 3-16 days) after endoscopic interventions. No difference was observed in efficacy and in time for the treatment of bile leak by sphincterotomy with endoprosthesis or endoprosthesis alone in patients with bile leak after surgery. CONCLUSIONS: Post-cholecystectomy bile leaks occur most commonly in the cystic duct and associated common bile duct stones are found in one-third of cases. Endoscopic therapy is safe and effective in the management of bile leaks and fistulae after surgery. Sphincterotomy with endoprosthesis or endoprosthesis alone is equally effective in the management of postoperative bile leak. PMID- 16698591 TI - Surgical therapy for hiliar cholangiocarcinoma: analysis of 198 cases. AB - BACKGROUND: Carcinoma of the hepatic duct confluence is the most common site of bile duct malignancies. Although hilar cholangiocarcinoma has been characterized as a slow-growing and late metastasizing tumor, post-therapeutic prognosis has remained poor. The study was undertaken to analyze factors influencing the surgical curative effect of hilar cholangiocarcinoma. METHODS: A retrospective clinical analysis was made of 198 patients with hilar cholangiocarcinoma who had been surgically treated at our hospital from 1997 to 2002. Jaundice (94.5%, 187 patients), pruritus (56.6%, 112) and abdominal pain (33.8%, 67) were the main symptoms. According to the Bismuth-Corlette classification, there were 14 type I patients, 19 type II patients, 12 type IIIa patients, 15 type IIIb patients, 112 type IV patients, and 26 unclassified patients. 144 patients received laparotomy, and 120 tumor resection including radical resection (59 patients) and palliative resection (61). Fifty-four patients were treated by endoscopic surgery and 16 patients by postoperative adjuvant radiation. RESULTS: Occupation, preoperative level of total serum bilirubin, operative procedure and postoperative adjuvant radiation affected postoperative survival of the patients. The postoperative survivals of endoscopic nose-biliary drainage (ENBD) group, endoscopic retrograde biliary drainage (ERBD) or endoscopic metal biliary endoprosthesis (EMBE) group, biliary exploration and drainage group, palliative resection group and radical resection group differed (chi2=87.0489, P<0.01). CONCLUSION: Early diagnosis and radical resection are important to improve the prognosis of hilar cholangiocarcinoma. PMID- 16698592 TI - Clinical analysis of patients with iatrogenic bile duct injury. AB - BACKGROUND: The main cause of iatrogenic bile duct injury is misidentification of the common bile duct as the cystic duct. In this article, we summarize the experience in the treatment of 112 patients with iatrogenic bile duct injury. METHOD: Clinical data of these patients treated at 10 hospitals of Songhua river area, Heilongjiang province, China from January 1978 to January 2005 were analyzed retrospectively. RESULTS: In 55.4% patients (62/112), iatrogenic bile duct injury was due to misidentification of the anatomy of Calot's triangle before cholecystectomy. Their diagnosis was based on clinical features, celiac puncture and imaging examination in which ultrasonography was most sensitive, giving a diagnostic rate of 97.5%. Six types of injury were identified according to their locations, and type III damage was commonly seen (92/112). The curative rate in this group was 95.5% (107/112). Eighty-seven patients (77.7%) underwent Roux-en-Y choledochojejunostomy with a cure rate of 94.3% (82/87). CONCLUSION: The prevention of iatrogenic bile duct injury lies in identifying the topography of extrahepatic bile ducts. Roux-en-Y choledochojejunostomy is usually the treatment of choice. PMID- 16698593 TI - Hepatobiliary membrane transporters involving in the formation of cholesterol calculus. AB - BACKGROUND: Cholecyst cholesterol lithiasis is a common disease of the digestive system; however, the cause of lithogenesis is still unclear. Although bile salt export pump (BSEP), multidrug resistance protein 2 (MRP2), and multiple drug resistance 3 (MDR3) are 3 well-known transporting proteins, their effect on lithogenesis has not been elucidated. This study was undertaken to examine the relationship between BSEP, MRP2, MDR3, and cholesterol calculus formation. METHODS: Liver tissue specimens were taken from 20 patients with cholesterol calculus and from 10 patients with normal liver. mRNA and protein expressions of BSEP, MRP2, and MDR3 were determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot, respectively. This study was approved by the ethics committee of China Medical University and informed consent was obtained from all patients. RESULTS: mRNA and protein expressions of BSEP, MRP2, and MDR3 were significantly down-regulated in the liver tissue of the patients with cholesterol calculus compared with normal liver tissue of the controls. CONCLUSION: The down-regulation of BSEP, MRP2, and MDR3 may be correlated with the formation of cholesterol calculus. PMID- 16698594 TI - Is there a role for serum endothelin in predicting the severity of acute pancreatitis? AB - BACKGROUND: Acute pancreatitis remains a common presentation to acute surgical units and carries significant morbidity and mortality. The progression of the disease to necrotizing pancreatitis and multi-organ dysfunction syndrome (MODS) is associated with a very poor clinical outcome, and persistently high mortality. Increases in serum endothelin (ET) have been seen in animal models of acute pancreatitis and this study aims to investigate whether there is a change in serum ET-1 in patients with acute pancreatitis and whether any such change is linked to disease severity. METHODS: All patients admitted with acute pancreatitis were prospectively recruited from the emergency admissions at the Norfolk and Norwich University Hospital. Serum ET levels were determined on admission, at 24 hours and 5 days post admission. Healthy adult controls were recruited from dermatology outpatients. RESULTS: A total of 21 patients joined the trial after giving informed consent. There were 3 men and 18 women with a median age of 65 years (range 26-87 years). Serum ET levels were significantly higher in acute pancreatitis patients than in normal controls (P<0.05). An association was seen between persistently raised serum ET levels and progression to MODS. CONCLUSIONS: The study does demonstrate a correlation between the circulating levels of ET and acute pancreatitis in humans, although it does not elicit its involvement in the pathogenesis of the disease. The observation that a persistently high level of circulating ET-1 is associated with progression to MODS may indicate a role for ET in the monitoring of acute pancreatitis patients for recovery or progression to MODS. PMID- 16698595 TI - APACHE system is better than Ranson system in the prediction of severity of acute pancreatitis. AB - BACKGROUND: It has been suggested that addition of obesity score to the APACHE-II system can lead to more accurate prediction of severity of acute pancreatitis. However there is scanty information on the usefulness of the combined APACHE-O scoring system in Asian patients. This study aimed to compare the accuracy of Ranson, APACHE-II and APACHE-O systems in assessing severity of acute pancreatitis in a local Chinese population. METHODS: One hundred and one consecutive patients with acute pancreatitis were prospectively studied. Body mass index (BMI) was measured on admission. Ranson score, APACHE-II and APACHE-O scores were recorded on admission and at 48 hours. By adopting the cut-off levels and definitions advocated in the Atlanta consensus for severe disease, the diagnostic accuracy of the three scoring systems was compared by the area under the curve (AUC) under the receiver operator characteristic curve. RESULTS: Of the 101 patients, 12 (11.9%) patients suffered from severe pancreatitis. Obesity was uncommon and only two patients (2.0%) had BMI>30. Eighty-two (81.2%) patients were normal weight (BMI< or =25) whereas 17 (16.8%) were overweight (BMI 25-30). Overweight or obesity (BMI>25) was not associated with severe pancreatitis (P=0.40). The AUC for admission scores of Ranson, APACHE-II, and APACHE-O systems was 0.549, 0.904 and 0.904, respectively. The AUC for 48-hour scores of Ranson, APACHE-II and APACHE-O systems was 0.808, 0.955 and 0.951, respectively. CONCLUSIONS: The APACHE-II scoring system is more accurate than the Ranson scoring system of the prediction of severity in acute pancreatitis. Addition of obesity score does not significantly improve the predictive accuracy of the APACHE-II system in our local population with a low prevalence of obesity. PMID- 16698596 TI - Solid-pseudopapillary tumor of the pancreas: CT and MRI features of 3 cases. AB - BACKGROUND: Solid-pseudopapillary tumor of the pancreas is a rather rare but low grade malignant tumor with good prognosis after surgical excision. METHODS: In 3 patients with solid-pseudopapillary tumor of the pancreas which were proved pathologically, the imaging features (CT in all and MRI in one patient) of the tumor were analyzed. RESULTS: In the 3 female patients under 35-year-old who presented no jaundice, CT revealed that huge solid-cystic masses of the pancreatic head with calcification in one and slight dilation of the pancreatic duct in another, but without dilation of the bile duct system in all. MRI demonstrated mixed signal intensity on T1WI and T2WI of the mass in one patient. CT and MRI revealed obvious enhancement of the cystic wall and solid part of the masses. CONCLUSIONS: CT and MRI findings of solid-pseudopapillary tumor of the pancreas are characteristic, and the diagnosis can be made preoperatively with the combination of clinical features. PMID- 16698597 TI - Breast carcinoma with metastasis to the gallbladder: an unusual case report with a short review of literature. AB - Gallbladder metastases are very rare and usually arise from malignant melanoma, renal cell carcinoma and cervical carcinoma. Breast carcinoma metastatic to the gallbladder is extremely rare and only 4 cases have been reported in the English literature. We hereby report a 54-year-old lady who was diagnosed as having breast carcinoma and underwent modified radical mastectomy. One month after the operation, she developed acute abdominal pain and underwent cholecystectomy after clinical investigation. Histopathological examination revealed metastasis to the gallbladder. Being considered a patient with metastatic breast carcinoma she was subjected to taxane and anthracycline-based palliative chemotherapy. Later she had CNS involvement and died of the progressive disease soon after few months. PMID- 16698598 TI - A case of portal vein thrombosis associated with acute cholecystitis/pancreatitis or coincidence. AB - BACKGROUND: Portal vein thrombosis (PVT) is complex and risk factors include local precipitating factors and acquired and inherited factors. It occurs secondary to abdominal malignancy, infection or surgical intervention. PVT is commonly forgotten as a possible cause of abdominal pain. The clinical picture may vary but abdominal pain and low grade fever are the most characteristic picture. METHODS: A 58-year-old male patient was admitted to our hospital complaining of abdominal pain for three days. CT scan revealed an edematous area around the portal vein. Doppler ultrasonography showed evidence of a portal vein thrombosis. RESULTS: PVT can be diagnosed with CT and Doppler ultrasonography. Fresh thrombus can be undetected by sonography because of the low echogenicity but can be recognized by color Doppler ultrasonography. Treatment ranges from observation and bowel rest to surgical resection of bowel. CONCLUSIONS: When we suspect a case of PVT, it should be treated at an early stage to prevent being lost in a diagnostic dilemma. The immediate use of anticoagulant could be important in preventing serious consequences of PVT. PMID- 16698599 TI - Diagnosis and management of extrahepatic portal vein aneurysm: a case report. AB - BACKGROUND: Extrahepatic portal vein aneurysm is a rare clinical entity with no more than 50 reported cases in the English literature. This study was to elucidate the procedures used in the diagnosis and management of portal vein aneurysm. METHOD: The clinical data of a patient with extrahepatic portal vein aneurysm were analyzed. RESULTS: Extrahepatic portal vein aneurysm associated with portal hypertension and splenomegaly was diagnosed by color Doppler ultrasonography (US), computed tomography (CT), magnetic resonance angiography (MRA) and angiography (indirect portography). Splenectomy and surgical shunt (splenorenal shunt) were performed. No hepatic encephalopathy and variceal bleeding occurred after the operation except mild pancreatic leakage caused by partial pancreatic tail resection. The patient recovered well. CONCLUSIONS: Extrahepatic portal vein aneurysm can be evaluated by US, CT, MRA, and angiography for optimum conservative or surgical management. PMID- 16698600 TI - Propofol is not so safe for ERCP. PMID- 16698602 TI - National Service Framework for long-term conditions--Quo vadis. AB - There is widespread dismay over the fate of the NSF for Long-term Conditions after all the efforts by so many people in securing its publication only one year ago in March 2005. In order to help to avoid its premature demise, the British Journal of Neurosurgery is pleased to provide a forum for the rapid publication of original papers and reviews that might help to catalyse the NSF's rational implementation. The first two of such papers are published in this Volume. PMID- 16698603 TI - A decade of change in regional head injury care: a retrospective review. AB - This paper reviews local and national changes and progress in the care of head injuries over the past decade in the Eastern Region. PMID- 16698604 TI - The spinal surgeon, the medical mishap, the warning and Afshar. AB - The patient suffering an injury and suing the doctor must establish a causal connection between the operation and the injury. If the doctor was not negligent in carrying out the operation, the patient can still succeed if the doctor did not inform and warn him of the predictable risks, however slight those risks might be, if they would or might have influenced the decision of the patient to consent or not. PMID- 16698605 TI - Characterization of cerebrovascular reactivity after craniectomy for acute brain injury. AB - Analysis of slow waves in arterial blood pressure (ABP) and intracranial pressure (ICP) has been used as an index to describe cerebrovascular pressure-reactivity. It has been previously demonstrated that the pressure-reactivity index (PRx) can be used to reflect global cerebrovascular reactivity with changes in ABP. A positive PRx signifies a positive association between ABP and ICP, indicating a non-reactive vascular bed, while a negative PRx is reflective of intact cerebral autoregulation, where ABP waves provoke inversely correlated waves in ICP. To date, there has been no characterization of pressure-reactivity following decompressive craniectomy. In this prospective observational study, 33 patients who underwent surgery for acute brain injury with mass lesions for which the bone flap was left out were studied. The PRx was calculated as a moving correlation coefficient between 30 consecutive samples of values of ICP and ABP averaged for a period of 10 s. The time profiles of mean PRx values at 6-hourly intervals were analysed and compared with that in seven patients treated by medical therapy alone. The initial mean PRx 6 h after surgery was positive, indicative of disturbed pressure-reactivity. With time, PRx trended towards a more negative value, suggestive of an improving cerebrovascular autoregulatory reserve. The mean PRx 24 h after surgery was 0.28 (+/-0.26), while the mean PRx 72 h after surgery was 0.15 (+/-0.25) (p = 0.012). In contrast, the mean PRx in patients that were not decompressed did not change significantly with time (p = 0.357). Surgery in acute brain injury for which the bone flap is left out in anticipation of raised intracranial pressure in the postoperative period leads to an improved PRx as compared with controls. Craniectomy in this situation may have a contribution to the restoration of disturbed cerebrovascular pressure-reactivity. PMID- 16698606 TI - Neurosurgical trainees operative experience before and after introduction of the New Deal for junior doctors. AB - To compare operative experience before and after implementation of the New Deal to reduce junior doctors working hours, operative data was audited over a 6-month period. The three registrars with national training numbers in our unit were placed on two virtual working patterns to determine what their operative experience would have been over that 6-month period. Comparison with a 1980s trainee over a similar period was also made. Trainees on a 1:3 rota would have performed more emergency operations [32 (28 - 38)], than those on a partial shift [1:6 with following day off, 16 (13 - 23)]. Particular examples include trauma craniotomies 13 (9 - 15) versus 6 (3 - 11) and CSF diversion, 9 (2 - 13) versus 4 (2 - 7). Although trainees on a 1:3 on call rota would have had more emergency operative experience over the same period of time, less common procedures, such as lumber decompression for cauda equina syndrome, were equally distributed between both groups. Trainees doing the partial shift would have lost 18 (12 - 24) elective cases over the same period as a result of enforced absence following periods of duty. These included posterior fossa, complex spinal, benign and rare cranial procedures. The increasingly common option of employing juniors on a full shift rota would have an even greater impact on training. Trainees are unavailable for elective training on night shift and then for a compensatory period afterwards, doubling the time spent away from formal surgical training. Indiscriminate reduction in working hours by enforced absence for compensatory rest has a potentially deleterious impact on elective training. By reducing the out of hours commitment trainees have a reduced, but possibly acceptable, exposure to emergency neurosurgery without impact on elective training. PMID- 16698607 TI - Inappropriate acute neurosurgical bed occupancy and short falls in rehabilitation: implications for the National Service Framework. AB - Patients undergoing neurosurgical intervention may require different types of organized rehabilitation. A prospective study was performed of the care needs of neurosurgical inpatients between the ages of 16 and 70 years who were in acute wards for more than 2 weeks. Only 58% of bed occupancy days were devoted to essential acute neurosurgical ward management. This figure was even lower for patients admitted with subarachnoid haemorrhage (36%) or traumatic brain injury (38%). Overall, 21% of bed days would have more appropriately spent in 'rapid access'/acute rehabilitation beds, 13% in 'active participation' rehabilitation beds and 5% in cognitive/behavioural rehabilitation units. Addressing this unmet need would increase the availability of acute neurosurgery beds, without needing to build and staff more neurosurgery wards. PMID- 16698608 TI - Craniotomy with prosthetic heart valves: a clinical dilemma. AB - The authors report two cases of spontaneous intracranial haemorrhage after elective craniotomy for resection of cerebral tumour. Both patients had mechanical aortic valve prostheses and were on regular warfarin therapy. In both cases, warfarin therapy was discontinued 5 days prior to surgery and unfractionated heparin administered intravenously until 12 h before surgery. Both patients were re-anticoagulated with subcutaneous low molecular weight heparin within the first week postcraniotomy-both developed life-threatening intracranial haemorrhage requiring urgent evacuation. The authors emphasize the risk of re anticoagulation without postoperative imaging and the disadvantages of therapeutic dose, low molecular weight heparin in the postoperative period. PMID- 16698609 TI - Awake craniotomy using stealth frameless stereotaxy without rigid skull fixation. AB - The authors describe a technique using the Medtronic Stealth spinal reference array allowing awake craniotomy to be performed without cranial fixation in the Mayfield pin head rest. A Medtronic spinal reference array (four-point H-shaped LED array) is fitted to a Yasargil footplate via a three-jointed swingarm. The Yasargil footplate is directly attached to the cranium after craniotomy and following stereotactic registration the patient is awakened. The patient is free to move his head during the procedure as the reference array does not move in relation to the cranial contents and the fiducials, preserving accuracy. PMID- 16698610 TI - Sciatic schwannoma spanning the sciatic notch: removal by an anterior, transabdominal approach. AB - We describe a sciatic schwannoma spanning the sciatic notch in a 39-year-old woman with persistent pelvic pain after caesarean delivery. The tumour was detected by pelvic CT scan and MRI. Anterior transabdominal surgery allowed the en bloc removal of both the pelvic and the buttock component of the tumour. PMID- 16698611 TI - Symptomatic syringomyelia secondary to clinically obscure infratentorial tumour. AB - The formation of a cervical spinal cord syrinx as a result of an infratentorial mass, even though uncommon, has been reported in international literature. In such cases, syringomyelia is usually asymptomatic, while the tumour-related symptoms and signs predominate. We report a patient with a posterior fossa tumour and secondary syringomyelia. In this patient, syringomyelia symptoms and signs were present, and a cervical spine Magnetic Resonance Imaging (MRI) showed a large cervical syrinx. A more careful clinical examination though, revealed a sub clinical posterior fossa syndrome and brain MRI revealed a large infratentorial meningioma. A posterior fossa craniotomy was performed, followed by complete tumour resection and almost complete remission of the syrinx and its related symptoms. The authors discuss the role of posterior fossa tumour induced tonsillar herniation in the development of secondary syringomyelia, the mechanisms leading to syrinx formation and the conditions that must be fulfilled for that to happen. PMID- 16698612 TI - Granular cell tumour of the neurohypophysis: a rare sellar tumour with specific radiological and operative features. AB - Symptomatic granular cell tumours of the neurohypophysis are rare sellar lesions. Preoperative prediction of the diagnosis on the basis of radiological appearance is useful as these tumours carry specific surgical difficulties. This is possible when the tumour arises from the pituitary stalk, rostral to a normal pituitary gland. This has not been emphasized previously. PMID- 16698613 TI - Emergency placement of a self-expandable covered stent for carotid artery injury during trans-sphenoidal surgery. AB - A patient sustained internal carotid artery (ICA) injury during trans-sphenoidal surgery. Bleeding from the resultant pseudo-aneurysm was not fully controlled by surgical packing. Emergency endovascular deployment over the injured ICA segment of a self-expandable covered-stent ('Symbiot' stent), initially designed for use in coronary saphenous vein-graft, was successful in securing haemostasis. PMID- 16698614 TI - Central neurocytoma in third and fourth ventricles with aqueductal involvement. AB - Central neurocytomas are characterized by their intraventricular locations, usually arising from the lateral ventricles and their benign clinical course. Variations in location, histology and clinical behaviour have been reported in recent years. The authors present two cases of central neurocytomas arising in the third and fourth ventricles with aqueductal involvement. The atypical features in their location and behaviour are discussed. The cases represent the extended spectrum of central neurocytomas. PMID- 16698615 TI - The patient always knows best! Conservative management of a large central disc prolapse. PMID- 16698620 TI - Screening of substance misuse during early pregnancy in Blyth: an anonymous unlinked study. AB - Substance abuse during pregnancy is rising and often remains undiagnosed. This harms both the mother and the baby. We conducted an anonymous unlinked study in Blyth valley, Northumberland, to determine the prevalence of substance abuse and alcohol use during pregnancy. Urine toxicology screening was performed on 150 women who attended antenatal clinic for booking. Seven commonly misused substances (amphetamine, benzodiazepine, barbiturates, cannabinoids, cocaine, methadone and opiates) and alcohol were tested in the urine. A total of 16 (10.7%) women were found positive and all 16 of them had denied use of any substance. Amphetamine was the most common among the substances misused. A total of 12 out of the 16 were from social classes 4 and 5. None of the positive women was nulliparous. Because of the difficulty in identifying substance misuse in pregnant women, consideration should be given to the implementation of routine substance screening at the booking antenatal visit in general population with a high incidence of substance misuse. PMID- 16698621 TI - Acceptability of prenatal HIV screening at the primary care level in Nigeria. AB - A survey of 333 pregnant women receiving antenatal care at the primary healthcare centres in Sagamu Local Government Area of Ogun State, southwest Nigeria was conducted between January and March 2005 to assess the acceptability of prenatal HIV screening among them. A total of 325 (97.8%) of the respondents were aware of HIV/AIDS but only 181 (54.3%) of them believed it is a problem in Nigeria. A total of 257 (77.2%) respondents agreed to undergo voluntary counselling and HIV testing (VCT). Multivariate logistic regression analysis of associated factors indicated that being married, self-perception of no risk of HIV infection, awareness of benefits of prenatal HIV testing and Christianity are independent predictors of acceptance of prenatal HIV testing in this population. Most of the respondents (78.9%) who were unwilling to take the test cited fear of being infected with its consequences of stigma and discrimination as the reason for their attitude. The survey suggests that a successful integration of VCT programme into the existing primary healthcare services for prevention of vertical HIV transmission is feasible in this part of Nigeria. PMID- 16698622 TI - Factors influencing gestational age at antenatal booking at the University College Hospital, Ibadan, Nigeria. AB - In order to achieve the aim of improved outcome of pregnancy for mother and fetus, early booking (first antenatal visit), prior to 14 weeks' gestation is usually recommended. A survey of information on personal data, index pregnancy, reasons for booking at a particular gestational age, past obstetric history and medical history of 205 pregnant women attending the antenatal booking clinic at the University College Hospital, Ibadan, Nigeria was carried out using a structured questionnaire. The mean gestational age at booking was 21.82 (+/-7.0) weeks. Only 29 patients (14.1%) booked before 14 weeks. The reasons given for early booking among them were the perceived benefits of such practice (41.4%), physician's recommendation (34.5%) and occurrence of complication(s) in previous pregnancy (24.1%). Illness in the index pregnancy and nulliparity were the only factors found to significantly favour early booking. The need to educate women of the reproductive age group, who are potential mothers, on the benefits of early booking was recommended. PMID- 16698623 TI - Utility of Doppler velocimetry in correlating with outcomes in severe discordant twins. AB - Twin birth weight discordance is associated with poor perinatal outcome. Three cases of severe discordant twins were serially assessed by ultrasound biometry and the Doppler ultrasonography of both the middle cerebral artery (MCA) and umbilical artery (UA) to measure pulsatility index (PI), resistance index (RI), and the systolic and diastolic ratio (S/D ratio). The birth weight discrepancies in three cases are 38%, 69% and 68%, respectively. Doppler velocimetry revealed a discrepancy in the S/D ratio of MCA and UA more than 15%. Absent end-diastolic flow, or the brain sparing effect, was noted in three small twins. Cerebral umbilical S/D ratio (CUR) (MCA/UA) of the twins was above 1 in large ones and below 1 in small ones. The small twin in case 3 expired after delivery. In conclusion, if the CUR of the S/D ratio of the fetus is below 1, the prognosis might be poor and fetal weight discordancy might be progressively larger. PMID- 16698624 TI - Obstetric complications and perinatal outcome in triplet pregnancies. AB - The incidence of multiple pregnancies with more than two fetuses has significantly increased since the introduction of ovulation agents and assisted reproductive technologies. Over a 15-year period there were 35 triplet pregnancies beyond 24 weeks that delivered at the King Fahad Hospital, an incidence of 1 in 1,099 deliveries. Early diagnosis is important for improving the rate of fetal salvage in triplet pregnancy. These pregnancies were managed on an outpatient basis. Prophylactic interventions were not utilised. A total of 91% of the pregnancies had at least one antenatal complication, pre-term labour being the most common (80%) followed by anaemia (43%). The average gestational age at delivery was 31.7 weeks (SD 4.2 weeks). A total of 94.3% of the patients were delivered by lower segment caesarean section. The mean birth weight of the neonates was 1,552 g (SD 510 g) and mean 5-min Apgar score was 7.6 (SD 0.8). The corrected perinatal mortality rate in the study was 152/1,000. Pregnancy outcome did not vary with birth order or mode of conception. Higher rate of pre-term births among triplet pregnancies make considerable demands on the neonatal intensive care unit. All methods of assisted reproduction should aim at prevention of multifetal pregnancies. PMID- 16698625 TI - Recurrent stillbirths: a matched case-control study of unexplained stillbirths at term. AB - The aim of this study was to assess the recurrent risk of an unexplained stillbirth at term. A total of 75 women who delivered stillbirths were matched for maternal age and parity with 75 controls. After excluding explained stillbirths, matched cases and controls were compared for maternal age, length of gestation, birth weight and 'interval to next birth'. The main outcome measure was the frequency of recurrence of a stillbirth. Both groups were similar for maternal age and length of gestation. Birth weight was marginally different (odds ratio (OR) = 0.997, 95% confidence interval (CI) 0.996, 0.999) and 'interval to next birth' was longer (OR = 1.08, 95% CI 1.00, 1.17). There were no stillbirths in cases and controls at follow-up. We conclude that a woman who has had an unexplained stillbirth at term has no greater risk of recurrence than a matched control. However, the 'interval to next birth' was significantly longer. PMID- 16698626 TI - Acceptability of caesarean delivery to antenatal patients in a tertiary health facility in south-west Nigeria. AB - Today, caesarean section is one of the most commonly performed surgical procedures the world over. Despite the well-documented record of safety, the strong aversion of women in sub-Saharan Africa to the procedure, especially in the presence of life-threatening indications, is of great concern to many obstetricians. This cross-sectional study, aimed at assessing the knowledge of the patients about caesarean section and its acceptability as mode of delivery, was conducted among antenatal patients at a University Teaching Hospital in south west Nigeria. A pre-tested structured questionnaire was used. Among the 201 patients surveyed, a high level of acceptability of caesarean section (85%) was found. However, 96.5% of those who would accept would give consent only after seeking the opinion of other people, especially their husbands. Previous major surgery and caesarean section were found to favour its acceptability, while age, tribe, marital status and the woman or her husband's educational status did not have any influence. With proper health education, especially during antenatal care, many more women would find caesarean section acceptable. PMID- 16698627 TI - Postpartum beliefs and practices in Danbare village, Northern Nigeria. AB - Postpartum cultural beliefs and practices are widely prevalent in northern Nigeria. Using a cross-sectional survey, we set out to examine contemporary postpartum beliefs and practices among a cohort of 300 mothers in Danbare village, northern Nigeria. Common postpartum practices included sexual abstinence (100%), physical confinement (88%), hot ritual baths (86%), nursing in heated rooms (84%) and ingestion of gruel enriched with local salt (83%). The majority of mothers (93%) believed that these practices made them stronger and helped them regain their physiologic state. Most respondents believed that non-observance could lead to body swelling, foul-smelling lochia and perineal pain. Mothers with formal education were significantly more likely to believe that these practices were non-beneficial compared with those mothers without formal education (odds ratio (OR) = 9.9, 95% confidence interval (CI) = 3.6 - 28.8). Almost half of the respondents (49%) said they would continue with these practices. In conclusion, women are still holding on to postpartum cultural beliefs and practices in northern Nigeria. However, educated women could act as useful agents of change towards the elimination of practices harmful to the health of mothers and their children. PMID- 16698628 TI - Problems associated with scuba diving are not evenly distributed across a menstrual cycle. AB - The problems encountered during scuba diving may be a contributing factor in an episode of decompression illness (DCI). Evidence exists that there may be a relationship between the position in the menstrual cycle and the occurrence of DCI. We examined, by prospective observation in female recreational scuba divers, any interaction between reported problems during diving (RPDD) and the position in the menstrual cycle. A total of 533 women, aged between 14 and 57 years, returned diaries for >6 months, with 61% returning diaries for 3 consecutive years. A total of 34,625 dives were reported within 11,461 menstrual cycles between 21 and 40 days in length, with 65% of women reporting at least one RPDD. Logistic regression showed a significant non-linear relationship between the position in the menstrual cycle and RPDD (p = 0.004). RPDD were not evenly distributed over the menstrual cycle; the rate per 1,000 dives varied from 39.2 at start of the cycle to 19.7 during week 3, and 31.9 in week 4. We concluded these field data suggest a possible correlation between the incidence of RPDD and the position in which they occurred in the menstrual cycle. PMID- 16698629 TI - Power Doppler in the assessment of pelvic masses in a low risk group. AB - Discrimination between benign and malignant adnexal masses aids in optimising the management of women with pelvic tumours. A grading system incorporating grey scale and power Doppler measurements might increase the accuracy of pre-operative assessment. The present study aimed to assess the sensitivity and specificity of a grading system including grey-scale and power Doppler findings. A total of 179 women who had undergone power Doppler Ultrasound for a pelvic mass between April 2001 and March 2003 were identified. Masses were surveyed by grey-scale and power Doppler and graded in relation to the estimated risk of malignancy. Clinicians were provided with the findings. The results were compared with the findings at surgery. A total of 74.3% were premenopausal. Some 94.4% (n = 169) had benign; 1.7% borderline and 3.9% malignant histology. A total of 69% underwent surgery. Specificity was 72.2 - 95.9%; sensitivity 55.6 - 88.9% and negative predictive value 97.6 - 99.2%. Power Doppler as an additional diagnostic aid may help the clinician in the management of pelvic masses. Further research is needed to clarify sensitivity and specificity. PMID- 16698630 TI - Adenomyosis: not such an elusive diagnosis any longer. AB - Adenomyosis has been an elusive diagnosis until recently due to the need for a histological confirmation of diagnosis post-hysterectomy. This commentary focuses on the role of non-invasive imaging techniques that have become available for use in the diagnosis of adenomyosis in women with an intact uterus. These include magnetic resonance imaging (MRI) and transvaginal ultrasound (TVS). With improvements in imaging modalities it has now become feasible to establish a diagnosis of adenomyosis without hysterectomy, to exclude additional pathology and to institute conservative symptomatic treatment and monitoring. PMID- 16698631 TI - Outcome of clinical pregnancies after ovulation induction using metformin. AB - The objective of this study was to review the first 50 clinical pregnancies of women with polycystic ovarian syndrome (PCOS) who had ovulation induced either with metformin alone, or in combination with clomifene. The study was confined to women with PCOS attending our infertility service. A register of clinical pregnancies was maintained of women who conceived after metformin therapy. The metformin was continued throughout the first trimester. The outcome of pregnancy was determined by individual chart review. Of the 50 women, 21 conceived with a combination of clomifene and metformin, and 29 with metformin alone. Seven women had a first trimester loss and 43 had a live birth. There were no perinatal deaths, no neonatal seizures and no congenital malformations. There were also no multiple pregnancies. The overall caesarean rate was 37%, and none of the babies had an Apgar score less than 7, at 5 min. This study found no evidence of any adverse clinical effects when metformin is continued in the first trimester of women with PCOS following ovulation induction. There was also no evidence of an increase in the rate of miscarriage or multiple pregnancy. PMID- 16698632 TI - Cetrorelix in women with poor ovarian response in previous assisted reproduction cycles. AB - Our aim was to compare a gonadotrophin-releasing hormone (GnRH) antagonist protocol with an analogue protocol using high dose gonadotrophins (rFSH) in women with poor ovarian response in order to optimise the management while undergoing assisted reproduction treatment. We recruited 31 consecutive patients over 5 months. The eligibility criteria for the study were: one or more previous cancelled cycle due to or=4,500 IU of rFSH. For the antagonist cycle regimen, we used daily 300 IU of rFSH from day 2 on the menses, and then from day 5 daily 0.25 mg of Cetrorelix until the day of human chorionic gonadotrophin (hCG) administration. We demonstrated that the use of an antagonist cycle was associated with a reduction in cancellation rates from 48% (agonist) to 10% (antagonist) (p < 0.039) allowing women to undergoing oocyte retrieval and embryo transfer with a non-significant improvement in the pregnancy rates. PMID- 16698633 TI - Development and psychometric testing of a symptom index for pelvic organ prolapse. AB - A wide range of symptoms are commonly ascribed to pelvic organ prolapse including pain, awareness of lump, bowel, bladder and sexual dysfunction. The aim of this work was to develop and validate an instrument to quantify symptoms related to pelvic organ prolapse. Consultation with symptomatic women and specialists in coloproctology, urology, gynaecology and sexual health resulted in a questionnaire with 25 questions. In total, 203 women participated in a psychometric testing of this instrument, 152 cases with prolapse and 51 controls without. The content validity, criterion validity, reliability and responsiveness of the questionnaire were evaluated. The questionnaire proved a reliable and valid instrument for the assessment of symptoms related to uterovaginal prolapse. It is also sensitive to change. PMID- 16698634 TI - A novel two-stage approach for the management of massive pelvic organ prolapse. AB - A new two-stage surgical procedure to treat massive pelvic organ prolapse in elderly women is described. Four women in whom previous pessary treatment had failed (two of these had also tried and failed the double ring pessary) or was not possible due to poor pelvic muscle tone or enlarged vaginal introitus underwent manual reduction of prolapse, colpoperineorrhaphy and ring pessary insertion in the first stage followed by a vaginal hysterectomy with or without sacrospinous fixation 6 weeks later. All patients had a favourable outcome in terms of improvement of symptoms and satisfaction with the treatment. PMID- 16698635 TI - A case of recurrent lymphocytic hypophysitis in pregnancy. PMID- 16698636 TI - Acute hepatic encephalopathy with acute pancreatitis in pregnancy. PMID- 16698637 TI - Myotonic dystrophy in pregnancy 'a salutary tale'. PMID- 16698638 TI - 'HELLP syndrome' in a patient with a negative pregnancy test. PMID- 16698639 TI - Cystic fibrosis diagnosed in late pregnancy. PMID- 16698640 TI - Spontaneous vaginal delivery through an unusual posterior cervical tear. PMID- 16698641 TI - Extra abdominal, intrauterine pregnancy. PMID- 16698643 TI - Avascular necrosis of femoral head associated with pregnancy. PMID- 16698642 TI - Application of the B-Lynch brace suture with associated intrauterine balloon catheter for massive haemorrhage due to placenta accreta following a second trimester miscarriage. PMID- 16698644 TI - An unusual case of haematocolpos with regular menstruation. PMID- 16698645 TI - Unusual cause of secondary amenorrhoea. PMID- 16698646 TI - Neglected vaginal ring pessary. PMID- 16698647 TI - Iatrogenic vesico-vaginal fistula caused by shelf pessary. PMID- 16698648 TI - Conservative treatment of an immature ovarian teratoma with gliomatosis peritonei. PMID- 16698649 TI - Ultrasound scan appearance of perforated uterus after surgical evacuation of retained products of conception. PMID- 16698650 TI - Successful use of TVT in the management of urodynamic stress incontinence in a treated case of carcinoma cervix. PMID- 16698651 TI - A large cystadenofibroma presenting in early pregnancy. PMID- 16698652 TI - Squamous cell carcinoma of the vagina: an unusual presentation. PMID- 16698654 TI - The pendulum swings back; estrogen is now beneficial if started at the right time. PMID- 16698655 TI - Homocysteine, estrogen and cognitive decline. AB - BACKGROUND: Factors that contribute to cognitive decline in women from midlife remain poorly understood. There are circumstantial data indicating a positive association between homocysteine and cognitive decline and that endogenous and exogenous estrogen may influence homocysteine levels. The aim of this review was to establish what is known of the relationships between cognitive change and homocysteine levels, and the impact of the menopause transition and exogenous estrogen on homocysteine levels. METHODS: We reviewed the recent published literature from 1993 to 2005 pertaining to the current understanding of the relationship(s) between plasma homocysteine levels and cognitive functioning and endogenous hormone levels and exogenous estrogen use in women. RESULTS: Hyperhomocysteinemia is consistently associated with cognitive decline. Dietary supplementation with vitamins may assist in normalizing homocysteine levels; however, there is no evidence that this results in favorable effects on cognition. Changes in endogenous estrogen levels are inversely associated with changes in serum homocysteine. Consistent with this, estrogen therapy is associated with reductions in plasma homocysteine, with the greatest effects reported in women with higher levels of homocysteine at baseline. Limited data indicate that tibolone is associated with little change in homocysteine. The use of raloxifene, the most studied selective estrogen receptor modulator, is associated with a modest reduction in homocysteine. CONCLUSIONS: There are data to suggest an underlying link between homocysteine levels and cognitive decline. There is also evidence for a link between both the menopause transition and use of exogenous estrogen therapy and homocysteine levels. Clinical data do not support a role for exogenous estrogen in the prevention of dementia in older women; however, the 'window of opportunity' theory suggests that there is a need for randomized controlled trials to evaluate the role of estrogen in the early postmenopausal years to protect against cognitive decline in later life. PMID- 16698656 TI - Hormones and progeny of breast tumor cells. AB - The rudimentary human glandular breast, with the approach of puberty, starts to grow both at glandular and stromal sites. Full differentiation is a gradual process and takes many years, and is only fully attained by pregnancy. The risk of breast cancer is inversely related to parity. Women during adolescence have the highest susceptibility to breast cancer development. This appears to be the period when the mammary gland has the highest number of stem cells. Stem cells may represent important targets for transformational events. Immunohistochemistry allows for identification of the lineage-specific precursor glandular and myoepithelial cells and their differentiated progeny. Both estrogen receptor subtypes are found in epithelial cells of alveoli and ducts as well as in stromal cells. Immunophenotypia of benign proliferative breast disease favors a fundamentally different epithelial composition from that of most malignant epithelial proliferations such as atypical ductal hyperplasia, ductal carcinoma in situ, lobular neoplasia and invasive breast carcinoma. Immunophenotypical characterization of these lesions assists in distinguishing benign from malignant disease. Based on the observation of bilateral risks and frequent multifocality with atypical ductal hyperplasia, atypical lobular hyperplasia and lobular carcinoma in situ, it is suggested that these may represent risk factors as well as precursors. One should, however, realize that ductal as well as lobular premalignant breast lesions ultimately arise from stem cells in the terminal duct lobular units. Estrogen receptor-beta (ERbeta)-positive and ERalpha-negative expression characterizes the highest levels of proliferative cancer cell activity. Point mutations and alterations of co-activators and co-repressors will also determine hormone sensitivity. There is evidence for different genetic pathways in the development of ductal carcinoma in situ and lobular carcinoma in situ. While they share recurrent 16q losses, a second hit in the E-cadherin gene explains the advent of lobular lesions and their common existence with the primary ductal type. Based on our immunocytochemical observations, the most likely target cell of malignant transformation is the Ck18/18-positive and ER negative transient cell of normal breast epithelium. Pregnancy confers a different genomic imprint to breast epithelial stem cells. Further elucidation of this mechanism may assist in developing appropriate means of breast cancer prevention. This paper is the Pieter van Keep Memorial Lecture given by Professor Hermann Schneider at the 11th World Congress on the Menopause, October 18-22, 2005, in Buenos Aires, Argentina. PMID- 16698657 TI - The long-term impact of 2-3 years of hormone replacement therapy on cardiovascular mortality and atherosclerosis in healthy women. AB - OBJECTIVE: The effect of hormone replacement therapy (HRT) on cardiovascular risk is intensely debated. The aim of this study was to investigate the long-term effects of HRT given for a few years on all-cause and cardiovascular mortality and the severity of atherosclerosis. METHODS: This analysis was based on a cohort of 1,458 postmenopausal women (55.8 +/- 6.1 years old) who previously participated in a number of randomized, placebo-controlled, clinical trials assessing the efficacy of 2-3 years of therapy with various estrogen plus progestin combinations for preventing bone loss. Women were followed on average for 9.8 years and came for a follow-up visit. Outcome variables were all-cause and cardiovascular mortality and the severity of atherosclerosis, as estimated by semi-quantitative scoring of vascular calcification in the lumbar aorta on lateral radiographs. RESULTS: A total of 174 women died during the observation period. All-cause mortality was decreased by 30% in the HRT+ group compared with the HRT- group (hazard ratio (HR) 0.70; 95% confidence interval (CI) 0.50-0.97) after adjusting for age, body mass index and smoking. Under the same conditions, similar results characterized mortality from cardiovascular disease (n = 61 deaths; 35.1% of all deaths) and coronary heart disease (n = 39 deaths; 22.4% of all deaths), which were decreased by 46% (HR 0.54, 95% CI 0.29-0.98, p = 0.045) and 53% (HR 0.47, 95% CI 0.21-1.03, p = 0.062), respectively. Furthermore, the mean severity score of aortic calcification at follow-up was significantly lower in hormone-treated compared to non-treated women (p < 0.0001). CONCLUSION: Women who receive 2-3 years of HRT after menopause do not have increased all-cause mortality, and results of the present study suggest relative cardiovascular benefits compared to those who had not used hormones. PMID- 16698658 TI - Quality of life and climacteric complaints amongst women seeking medical advice in Taiwan: assessment using the WHOQOL-BREF questionnaire. AB - OBJECTIVE: To determine the impact of the climacteric transition on health related quality of life amongst women, between the ages of 45 and 55 years, seeking medical advice in Taiwan. METHOD: A total of 203 women seeking medical advice (SMA) were drawn from a special integrated clinic, with a further 349 healthy referents of the same age, range and gender, with no history of hormone replacement therapy and living in the same municipality, also being recruited from a national health survey sample for comparison. Each one was asked to fill out the brief questionnaire of the Taiwan version of the World Health Organization Quality of Life (WHOQOL-BREF), assessing quality of life on 26 items in four domains (physical, psychological, social and environmental). SMA subjects were also questioned about the 21 most frequent symptoms. Multiple regression analyses were conducted to control variables such as age, marital status, religion and educational attainment. RESULTS: The mean scores for the physical, psychological and social domains were significantly lower than those of the healthy referents, as was the overall quality of life for SMA women. Although usual vasomotor symptoms did not significantly predict quality of life in the SMA subjects, after controlling for demographic factors, insomnia and emotional disturbance were found to be major determinants of the scores in the different domains. CONCLUSION: Insomnia and emotional disturbance should be taken into consideration in the management of climacteric women seeking medical advice. PMID- 16698659 TI - Efficacy and acceptability of risedronate 5 mg daily compared with 35 mg once weekly for the treatment of postmenopausal osteoporosis. AB - OBJECTIVE: Risedronate is a bisphosphonate used in the treatment of osteoporosis. It has a strong effect in inhibiting osteoclast activity. The primary objective of this study was to evaluate the effectiveness and adverse events of two different forms of risedronate (5 mg and 35 mg) using a rapid biochemical marker for comparison of C-terminal telopeptide (CTx) type I collagen cross-links. METHODS: The study was designed at Bakirkoy Dr. Sadi Konuk Education & Research Hospital, between January and June 2004. A total of 123 postmenopausal osteoporotic women were randomly assigned in blocks of three to one of the following groups: control, risedronate 5 mg/day and risedronate 35 mg once a week. RESULTS: Of the 123 women enrolled, 103 (83.7%) completed the study. Adverse events were experienced by 53.6% in the control group, 56% in the risedronate 5 mg/day group and 53.6% in the group receiving risedronate 35 mg once per week. The most common adverse events were gastrointestinal (21.9% of subjects in group 1, 29.2% of subjects in group 2, 24.3% of subjects in group 3). The women in groups receiving risedronate either 5 mg/day and 35 mg once per week had similarly decreased levels of CTx but the control group was not as effective. CONCLUSION: The results support the hypothesis that risedronate 35 mg given once per week has the same therapeutic efficacy and safety as a daily 5 mg regimen. Taking the medicine once a week is likely to be easier and more satisfactory than the daily regimen. However, patients taking a once-a-week regimen may forget to take it due to the 7-day break without medicine. PMID- 16698660 TI - The influence of premenopausal hormones on severity of climacteric symptoms and use of HRT. AB - OBJECTIVES: There is wide variation in the severity of climacteric symptoms and we hypothesized that this could be a reflection of premenopausal hormone levels. METHODS: As part of a long-term cohort study of endocrine risk factors for breast cancer, blood had been collected between 1986 and 1990 from 1882 premenopausal women aged >or=35 years. Questionnaires on menopausal symptom severity were sent to 1,843 surviving women in 2001, of whom 1,434 replied. Estradiol, progesterone and testosterone levels were measured by radioimmunoassay in 680 women who reported a natural menopause and completed the symptom severity section in full. RESULTS: Symptom severity fell with rising premenopausal estradiol levels and women with higher premenopausal testosterone levels had more severe vasomotor symptoms. Over 70% of women with above-median severity of symptoms had used hormone replacement therapy (HRT). Those with higher testosterone levels were less likely to take HRT. CONCLUSIONS: Premenopausal hormone levels may predict risk of severe menopausal symptoms, which in turn influences use of HRT. Paradoxically, a high testosterone level was associated with more vasomotor symptoms but reduced use of HRT. Those at greatest risk of climacteric symptoms may be at lower risk of breast cancer because of premenopausal reduced estrogen exposure. PMID- 16698661 TI - Guidelines for hormone replacement therapy of Asian women during the menopausal transition and thereafter. AB - These Guidelines summarize the position of an Expert Panel on Menopause in Asian Women regarding the use of hormone replacement therapy (HRT) during the menopausal transition and thereafter. They are intended to aid gynecologists, family physicians and other health-care professionals in providing optimal care to menopausal Asian women who desire HRT. PMID- 16698663 TI - Effects of adenovirus-mediated gene transfer of ICOSIg and CTLA4Ig fusion protein on experimental autoimmune myocarditis. AB - OBJECTIVE: To explore the therapeutic alliance effects of adenovirus vector mediated gene transfer of ICOSIg and CTLA4Ig fusion protein on experimental autoimmune myocarditis (EAM). METHODS: Expression vector pAdeno-CTLA4Ig and pAdeno-ICOSIg was constructed and transfected into HEK293 cells. Adenovirus expresses CTLA4Ig and ICOSIg was produced. Ad-CMV-GFP was used as controls. EAM was induced in Lewis rats by injection of procine cardiac myosin. All the immunized rats were divided into four groups. Group A (n = 15) received adenovirus containing CTLA4Ig and ICOSIg from day 14-28; group B (n = 15), group C (n = 15) and group D (n = 15) received adenovirus containing CTLA4Ig, ICOSIg and GFP, respectively. Group E (n = 10) was normal controls never received immunization. On day 28, all the rats were killed after echocardiography examination. Histopathological examination was used to observe inflammation in the myocardium. Western blot was used to detect CTLA4, ICOS, ICOSL and competitive RT-PCR for B7-1, B7-2 expression. T lymphocyte proliferation assay was performed and ELISPOT was used to detect the Th1 and Th2 production. RESULTS: Alliance application of CTLA4Ig and ICOSIg exerts therapeutic effects on EAM. After a treatment duration of 14 days, cardiac function and myocardial inflammation improved significantly compared to group D. Expression of CTLA-4, ICOS and ICOSL, B7-1 was statistically decreased in group A, B and C compared with group D. T-cell proliferation was inhibited by costimulatory blockade in a dose-dependent style. ICOSIg blockade significantly augments IL-4 and IL-10 production while diminished IFN-gamma production. CONCLUSIONS: Blockade of costimulatory pathway with alliance therapy of CTLA4Ig and ICOSIg alleviated autoimmune damage in EAM and improved cardiac function. The mechanisms may be downregulation of costimulatory molecules and anti-inflammation. PMID- 16698664 TI - A reduced IL2R (CD25) expression level in first and second degree female relatives of autoimmune thyroid disease patients. A sign of a poor capability to preserve tolerance? AB - There is room for immune markers other than TPO-Abs to identify an increased risk to develop autoimmune thyroid disease (AITD). Our aim was to test the hypothesis that activation of CD4+ T cells is such marker in relatives of AITD patients, who have an increased risk to develop AITD. We established a controlled study on 20 TPO-Ab positive and 20 TPO-Ab negative euthyroid female relatives. All these cases had at least one 1st or 2nd degree relative with a documented autoimmune hyper- or hypothyroidism in whom we studied the percentages of circulating subsets of activated (MHC class-II, CD25 (IL-2R), CD71 or CD69+) CD4+ T cells and the level of the soluble (s)-IL2R in serum. We found that euthyroid female relatives did not show an activation of their T cell system, but a reduced expression of CD25 on CD4+ T cells. The level of the shed IL2R in serum was also lower in comparison with levels found in healthy control females. A reduced T cell activity was found in both TPO-Ab positive and negative relatives. In conclusion, female relatives with at least one 1st or 2nd degree relative with an AITD show signs of a reduced expansion capability of their T cell pool. It is hypothesized that this reduced expansion capability may affect T cell tolerance mechanisms more than T effector mechanisms. PMID- 16698665 TI - Identification of nuclear spliceosomal antigens targeted by NOD mouse antibodies following sodium iodide intake. AB - The non-obese diabetic (NOD) mouse spontaneously develops a range of autoreactive responses including an autoantibody response to nuclear antigens. As elevated dietary iodine has been shown to increase thyroid autoimmune pathology in NOD mice, the effect of sodium iodide (NaI) on the development of anti-nuclear antibodies (ANA) was assessed. Interestingly, the NaI symporter is expressed in both thyroid and salivary glands. Elevated dietary iodine was found to increase the percentage of male NOD mice developing autoantibodies. Specifically, the nuclear autoantibodies that develop in NOD mice were shown to target specific spliceosomal components. The target specificity of the autoantibodies was determined using recombinant spliceosomal proteins and shown to include U1A, U170K, U2B'', U2A', as well as the Sm proteins D1, D2, and B. The autoantibody isotypes most consistently represented were IgG2a and IgG2b. PMID- 16698666 TI - IgA class antibodies to 2-oxo-acid dehydrogenase complex are not predictive markers of histopathological progression in primary biliary cirrhosis. AB - Although antimitochondrial antibody (AMA) is the characteristic serological feature of primary biliary cirrhosis (PBC), its pathogenic role remains unclear. In our previous study, we reported a positive correlation between immunoglobulin (Ig) A class anti-2-oxo-acid dehydrogenase complex (2-OADC) and histopathological stage. To determine whether the appearance of IgA class anti-2-OADC by immunoblotting represents an early marker of more aggressive disease or whether it is late finding during the disease course of PBC, we tested not only the entire IgA class but also IgA1, IgA2 and secretory IgA class anti-2-OADC in serial serum samples from 15 patients with PBC. During the median observation period of 51 months, four cases showed histopathological progression (from stage 1 to 2, stage 1 to 3, stage 1 to 4 and stage 2 to 4). There was no statistically significant correlation between the above IgA class anti-2-OADCs and histopathological progression. There was no significant correlation between histopathological stages and IgA2 class anti-2-OADC or secretory IgA class anti-2 OADC by immunoblotting. IgA class anti-2-OADC was more frequent in stages 3-4 than in stages 1-2 (p = 0.0049), but IgA1 class anti-2-OADC was more frequent in stages 1-2 than in stages 3-4 (p = 0.0232). Our present study demonstrated that serum IgA class 2-OADC was not a predictive marker of histopathological progression but was associated with the histopathological stage of PBC. Although the IgA class AMA may have a specific pathogenic role for PBC, the discrepant results between IgA and IgA1 class anti-2-OADC should be further assessed to investigate different functional activities depending on their molecular form. PMID- 16698667 TI - A new oligonucleotide-based ELISA for the detection of anti-double-stranded DNA antibodies. AB - The detection and measurement of antibodies to double-stranded (ds) DNA is important in the diagnosis and monitoring of patients with systemic lupus erythematosus (SLE). Several methods are available for their determination but none of them is completely satisfactory. Usually, purified dsDNA from different sources is used as antigen in the solid (enzyme-linked immunosorbent assay, ELISA) or liquid phase (Farr assay). Alternatively, anti-dsDNA antibodies can be demonstrated by immunofluorescence, using the haemoflagellate Crithidia luciliae (CLIFT). We have developed a new oligonucleotide-based anti-dsDNA ELISA in which dsDNA, constituted of a 5'photobiotinylated nucleic acid probe and its highly specific complementary oligonucleotide, is formed onto the solid-phase. To do that, a 40 bp probe is coated on the microplate wells via streptavidin-biotin bond and the in vitro (in solid-phase) developed hybrid between probe and its complementary helix used as capturing antigen. The assay has proved to be specific and the several experiments performed to ascertain the absence of single stranded DNA (ssDNA) contamination and/or non specific binding to plastic, streptavidin, probe (without complementary chain) have all excluded any significant interference. To evaluate the clinical performance of this new assay, 114 serum samples from 68 SLE patients and 85 samples from 85 disease controls, in addition to 30 blood donors, were studied. The results were compared with commercial ELISAs, Farr assay and indirect immunofluorescence. The sensitivity and specificity were 66.2 and 96.4%, respectively, comparable and even better than those of the commercially available anti-dsDNA kits. Anti-dsDNA antibody levels, measured with the oligonucleotide-ELISA, correlated with SLE clinical activity determined using the European Consensus Lupus Activity Measurement (ECLAM) (r = 0.59, p < 0.0001). In conclusion, this assay has proved to be reproducible, and the results correlate well with other standard anti-dsDNA assays. The features of this new assay make it promising for clinical use. PMID- 16698668 TI - Transfer of immune components from rabbit autoimmune cardiomyopathy into severe combined immunodeficiency (SCID) mice induces cardiomyopathic changes. AB - BACKGROUND: Growing evidence suggests that autoimmune mechanism plays an important role in the pathogenesis of cardiomyopathy. The purpose of this study was to investigate whether passive transfer of IgG and/or lymphocytes from rabbits with autoimmune cardiomyopathy is able to reproduce cardiomyopathic changes in severe combined immunodeficiency (SCID) mice. METHODS AND RESULTS: SCID mice were injected intraperitoneally with IgG and/or peripheral blood lymphocytes (PBL) from either rabbits immunized with both beta1-adrenoceptor peptide and M2-muscarinic receptor peptide (beta1+M2 group) or rabbits with adjuvant (N group). Thirty five SCID mice were divided into seven groups; N-IgG, N-PBL, N-IgG & PBL, (beta1+M2)-IgG, (beta1+M2)-PBL, (beta1+M2)-IgG & PBL and control groups. Heart weight in three (beta1+M2) groups were significantly increased. All mice in three (beta1+M2) groups showed high titer of rabbit anti beta1 adrenoceptor autoantibodies, and 4 mice in the (beta1+M2)-PBL group and 3 mice in the (beta1+M2)-IgG & PBL group showed a significant increase in titer of rabbit anti-M2-muscarinic receptor autoantibodies. Focal infiltration of inflammatory cells in the myocardium was observed in the (beta1+M2)-IgG & PBL group. In the (beta1+M2)-PBL group and (beta1+M2)-IgG & PBL group, cardiomyocyte diameters were significantly increased. Some myocytes of the (beta1+M2)-IgG & PBL group exhibited intracellular edema, clumps of Z-band and increased numbers of mitochondria by using electron microscopy. CONCLUSION: Transfer of IgG and PBL from rabbits immunized with combined beta1 and M2 peptides was able to reproduce the early stage of cardiomyopathic changes in SCID mice. PMID- 16698669 TI - Lipoteichoic acid may affect the pathogenesis of bile duct damage in primary biliary cirrhosis. AB - AIM: Intrahepatic bile ducts are the targets for inflammation in primary biliary cirrhosis (PBC), but their pathogenesis is not known. Gram-positive bacterial DNA was detected recently in gallbladder bile of PBC patients. In the present study, we assessed the possible pathological role of lipoteichoic acid (LTA), the gram positive bacterial cell wall component, in PBC. METHODS: Liver samples, obtained from 20 patients with PBC (stage 1-2 with CNSDC: stage 3-4 with loss of bile ducts = 10:10) and from 13 patients with chronic hepatitis due to hepatitis C virus (CH-C) with lymphocytic cholangitis, were subjected to immunohistochemical staining with polyclonal rabbit anti-LTA as the primary antibody. Serum reactivities to LTA were studied by ELISA. After 1 microg of purified LTA was placed in a 96-well microplate as an antigen, an antibody capture assay was carried out using serum samples from PBC (n = 20), CH-C (n = 13) and healthy subjects (n = 11). RESULTS: LTA was localized around the sites of chronic non suppurative destructive cholangitis (CNSDC) in the portal area in stage 1-2 PBC but was not detected in the portal area in CH-C. In stage 3-4 PBC, LTA was localized around sites of ductular proliferation at the periphery of portal tracts. IgM class anti-LTA serum titers were significantly higher in PBC than in CH-C. IgA class anti-LTA serum titers were significantly higher in PBC than in healthy subjects. CONCLUSIONS: In the PBC livers, the profile of immunoreactivity to LTA changed markedly as the disease progressed. Sera from PBC showed higher levels of anti-LTA titers than CH-C (IgM) or from healthy subjects (IgA). The LTA mediated immune system might affect the initiation and/or progression of PBC. PMID- 16698670 TI - Reduced tetanus antibody titers in overweight children. AB - Under-nutrition impairs immune responses, but far less is known about the impact of over-nutrition, such as obesity, on the response to vaccines. We measured the effect of childhood overweight status on inflammatory mediators, circulating immunoglobulins and tetanus antibodies in fifteen overweight children (BMI > 85 age-adjusted percentile) and 15 age-matched normal weight controls. Fitness was measured by a progressive ramp type exercise test. Lean body mass (LBM) and fat mass were determined by DXA. Tumor necrosis factor-a (TNF-a), interleukin-6 (IL 6), interleukin-1 beta (IL-1beta) and interleukin-1 receptor antagonist (IL-1ra) were used to assess the inflammatory status; and circulating immunoglobulins (IgM, IgA, IgG and IgG subclasses) and specific IgG titer to tetanus were used to assess humoral immunity. Overweight children had higher LBM and percent fat mass, and lower peak VO2 normalized to body weight. IL-6 was significantly higher in the obese children (2.6 +/- 0.3 vs. 1.3 +/- 0.3 pg/ml, in overweight and normal weight children, respectively; p < 0.05). No significant differences were found in TNF-a, IL-1beta and IL-1ra between the groups. No significant differences were found in immunoglobulin levels (IgM, IgA, IgG and IgG subclasses) between the groups. Anti-tetanus IgG antibodies were significantly lower in the overweight children compared to normal weight controls (2.4 +/- 0.6 vs. 4.2 +/- 0.5 IU/ml, in overweight and normal weight children, respectively; p < 0.05). The reduced specific antibody response to tetanus in obese children and adolescent might be due to mechanical factors such as lower relative vaccination dose, or reduced absorption from the injection site due to increased adipose tissue, or related to reduce immune response due to the chronic low grade inflammation expressed by the higher levels of IL-6. PMID- 16698671 TI - Cannabidiol lowers incidence of diabetes in non-obese diabetic mice. AB - Cannabidinoids are components of the Cannabis sativa (marijuana) plant that have been shown capable of suppressing inflammation and various aspects of cell mediated immunity. Cannabidiol (CBD), a non-psychoactive cannabidinoid has been previously shown by us to suppress cell-mediated autoimmune joint destruction in an animal model of rheumatoid arthritis. We now report that CBD treatment significantly reduces the incidence of diabetes in NOD mice from an incidence of 86% in non-treated control mice to an incidence of 30% in CBD-treated mice. CBD treatment also resulted in the significant reduction of plasma levels of the pro inflammatory cytokines, IFN-gamma and TNF-alpha. Th1-associated cytokine production of in vitro activated T-cells and peritoneal macrophages was also significantly reduced in CBD-treated mice, whereas production of the Th2 associated cytokines, IL-4 and IL-10, was increased when compared to untreated control mice. Histological examination of the pancreatic islets of CBD-treated mice revealed significantly reduced insulitis. Our results indicate that CBD can inhibit and delay destructive insulitis and inflammatory Th1-associated cytokine production in NOD mice resulting in a decreased incidence of diabetes possibly through an immunomodulatory mechanism shifting the immune response from Th1 to Th2 dominance. PMID- 16698672 TI - The autoimmune characteristics of progressive or sudden sensorineural hearing loss. AB - Autoimmune sensorineural hearing loss (ASNHL) is a clinical syndrome that typically produces a bilateral rapidly progressive hearing loss. Autoantibodies and autoreactive T cells have been implicated in the etiopathogenesis of ASNHL. However, the identity of a specific or highly relevant inner-ear self-antigen is still required. In recent years, a role for antiphospholipid antibodies has been also suggested, assuming sudden/progressive hearing loss to be part of the primary antiphospholipid syndrome (APS). In this review, we address the need for specific diagnostic tools in order to establish an autoimmune origin for hearing loss. Moreover, the decision of whether anti-inflammatory drugs or anti coagulation should be given is also discussed. PMID- 16698673 TI - Use of alum and inactive Bordetella pertussis for generation of antibodies against synthetic peptides in mice. AB - We have investigated the efficacy of the combined use of Alum and inactive Bordetella pertussis (iBP) adjuvants for eliciting anti-peptide antibodies. ICR mice were immunized four times at 3-week intervals with each of 7 free (i.e., not conjugated to any carrier) synthetic peptides of 15-17 amino acid residues in Alum + iBP, in the commonly used adjuvant protocols (CFA; CFA (initial) followed by IFA), or in CFA + iBP. Serum samples after 3 and 4 injections were tested by RIA. Use of Alum + iBP greatly increased the production of antibodies for most of the peptides. The results have important implications for human vaccine formulation involving peptides. PMID- 16698674 TI - Sphingosine kinase and sphingosine-1-phosphate regulate migration, endocytosis and apoptosis of dendritic cells. AB - Dendritic cells (DC) are inducers of primary immune responses and represent an attractive vector for cancer immunotherapy. Sphingosine kinase (SphK) and its product sphingosine-1-phosphate (S1P) play an important role in the regulation of immune cells and cancer, affecting processes such as differentiation, growth or migration. We studied the role of SphK and S1P on migration of DC. RT-PCR showed mRNA expression of SphK in DC, declining from immature (iDC) to mature DC (mDC) to antigen-loaded mDC. Expression of S1P receptors was S1P(1) > S1P(2) = S1P(3), unrelated to maturation or antigen uptake. In transwell assays, iDC migrated towards SDF-1, MIP-1alpha, MCP and S1P, whereby S1P combined with a chemokine had a synergistic effect. mDC migrated towards 6Ckine and MIP-3beta, but not towards S1P. The SphK-inhibitor dihydro-sphingosine (DHS) reduced migration of iDC but not of mDC. In addition S1P(3)-inhibitor suramin inhibited DC migration in response to S1P. DHS had a reverse effect on endocytosis, enhancing the uptake of FITC dextran. We also observed an anti-apoptotic effect of S1P on mDC for the first time. This indicates that SphK/S1P may play a role in accumulation of peripheral iDC at the location of antigen and subsequent antigen-uptake. These findings may help to optimise DC-based cancer immunotherapy by modulation of SphK/S1P. PMID- 16698676 TI - Description of a novel panallergen of cross-reactivity between moulds and foods. AB - The present investigation is undertaken to demonstrate a novel cross-reactivity between aeroallergens (moulds fungi imperfecti) and allergens from foods (spinach and mushroom Agaricus bisporus). We have performed a dual study in vivo and in vitro, in a population of atopic patients. Data from in vivo tests performed with spinach and mushroom have been statistically analysed. To the in vitro assays, mushroom and spinach extracts have been obtained, and sera from moulds allergic patients analysed by means of IgE-immunoblott assays. Inhibition experiments have been also performed to study a possible relation between proteins. Statistical analysis of data showed a relation between allergenicity to moulds (Alternaria alternata, Cladosporium herbarum and/or Aspergillus fumigatus), and positive skin prick tests with mushroom and/or spinach. The immunoblotts performed showed that seven moulds allergic patients had a strong recognition of a protein with a molecular weight of about 30 kD present both in spinach and mushroom extracts, and by means of inhibition assays we could determine that these two proteins were related. This study demonstrates the existence of a new allergen responsible for cross reactivity between moulds and two frequently consumed foods, mushroom and spinach. We conclude that a novel cross-reactive allergen between aeroallergens and foods has been identified. PMID- 16698675 TI - MPO and cytokines in the serum of cancer patients in the context of Candida colonization and infection. AB - This study investigated the immunological factors, such as neutrophils number, the level of myeloperoxidase and IL-12, IL-10, TNF-alpha, IFN-gamma, that additionally might correlate with increased susceptibility to Candida infections in cancer patients. A total of 105 cancer patients were evaluated. Patients were examined twice for Candida colonization and presence of Candida antigen and DNA in bloodstream. Serum concentrations of MPO and selected cytokines were quantified by ELISA. The values for myeloperoxidase were decreased in Candida colonized as well as deep-infected cancer patients groups, compared to healthy persons. In the group of patients suspected of deep candidiasis, we observed significantly elevated level of IFN-gamma compared to control. In the group of Candida-colonized patients, the concentrations of IL-12, TNF- alpha and IFN-gamma were significantly heightened when compared to control.MPO deficiency seems to be one of the important risk factor for deep candidiasis independently of the neutrophil count. The disturbances in cytokines levels in cancer patients group can be connected with underlying cancer disease, its treatment as well as Candida infection. The decreased level of TNF-alpha, in particular may be connected with Candida invasion. PMID- 16698677 TI - The role of IL-4 and IL-12 in the regulation of collagen synthesis by fibroblasts. AB - Chronic sclerodermifomic graft versus host disease is a rare but important complication of allogeneic hematopoietic stem cell transplantation that especially occurs in patients who are treated with donor lymphocyte infusions for relapse of a malignant disease. Today most knowledge about the pathogenesis of chronic Graft-versus-Host Disease is based on mice models. In this report we describe the development of an allogeneic in vitro model that allows studying the pathogenesis of chronic sclerodermifomic Graft-versus-Host Disease in the human setting. We report that priming of mononuclear cells in the presence of allogeneic fibroblasts and Interleukin (IL)-4 induces fibroblast collagen synthesis, whereas priming in the presence of IL-12 suppresses collagen synthesis during subsequent coculture of primed mononuclear cells with allogeneic fibroblasts. Since IL-12 is also known to mediate anti-tumor effects by stimulation of Natural Killer cell and Lymphokine Activated Killer cell activity, these findings indicate that treatment of patients with IL-12 or pretreatment of donor lymphocytes with IL-12 might strengthen a graft versus leukemia effect and at the same time decrease the risk of chronic sclerodermifomic Graft-versus-Host Disease development. PMID- 16698678 TI - Cord blood-derived plasma factor (CBPF) potentiates the low cytokinetic and immunokinetic profile of bone marrow cells in pesticide victims suffering from Acquired Aplastic Anaemia (AAA): an in vitro correlate. AB - Cases of Acquired Aplastic Anaemia (AAA) in patients with a long history of pesticide exposure from agricultural fields have been investigated in our laboratory using an immunological approach. These patients showed moderate to severe degrees of bone marrow aplasia as a result of 9-12 years protracted exposure to pesticides which were mainly comprised of organophosphorous and organochloride compounds. The bone marrow aspirate culture was found to be severely deficient both in terms of differentiation and proliferation, and cell mediated immune function (CMI). We attempted ex vivo manipulation of the bone marrow population of patients in two different protocols: in one, stem cell factor (SCF), interleukin-3 (IL-3), and granulocyte-colony stimulating factor (G CSF) were administered and, in the second set, cord blood-derived plasma factors (CBPF) were supplemented to evaluate the effects, if any. Simultaneously, two control groups including one for healthy normal control (N) and the second, for non-pesticide induced aplastic anaemia group of patients (NPAA) was also investigated for all the above parameters. Active colony formation and improved cellular immune activity (CMI) was observed more frequently in the CBPF treated group rather than that in the cytokine treated group. Surprisingly, administration of cytokines in the first set and CBPF in the second set triggered CD34 (+) cell generation as revealed through flow cytometric analysis (FACS). The effect was more pronounced in the second set. Investigations carried out with NPAA showed relatively insignificant effects with both cytokine and CBPF set up. The investigations indicated that AAA as induced by pesticides could be therapeutically manipulated by exogenous cytokines and growth factors and, more efficiently, by CBPF by way of immunopotentiation through microenvironmental supplementation. PMID- 16698679 TI - Investigation of GM-CSF immune accessory effects in tumor-bearing mice by direct gene immunization. AB - To assess GM-CSF immune accessory effects in tumor-bearing mice, an animal tumor model was established by inoculating SP2/0 myeloma cells s.c. into the flank of Balb/c mice and 14 days later, injecting either 400 mug recombinant pcDNA3.1/mGM CSF or a blank plasmid s.c. or i.m. into the tumor four times. The tumor weight, the activities of CTL and NK, the serum levels of IFN-gamma, IL-2 and lymphocytes infiltrating in tumor tissue were analysed 8 weeks later with MTT, ELISA and pathological section methods. The results showed that the tumor lump was reduced in mice injected s.c. (0.880 +/- 0.405 g) or i.m. (0.378 +/- 0.411 g) with pcDNA3.1/mGM-CSF compared with control mice injected s.c. (1.548 +/- 0.221g, P < 0.01)or i.m. (1.554 +/- 0.249g, P < 0.001) with a blank vector. Lymphocyte infiltration in tumor tissues was very apparent in mice injected i.m. with pcDNA3.1/mGM-CSF. In contrast, there was no lymphocyte infiltration in tumor tissues of control mice. In addition, the serum concentrations of IFN-gamma, IL-2 and the activities of CTL and NK cells were significantly increased in mice injected with pcDNA3.1/mGM-CSF compared with a control mice (P < 0.01). In conclusion, direct gene immunization of recombinant pcDNA3.1/mGM-CSF is a feasible strategy for tumor therapy. PMID- 16698680 TI - Polymorphism in the promoter region of the CD14 gene and susceptibility to Brucellosis. AB - A single-nucleotide polymorphism in the promoter region of the CD14 gene at position 159 has been implicated in susceptibility to infectious diseases. We sought to determine the association between CD14 C-159 T functional promoter polymorphism and brucellosis in Western Iranian population where the disease is endemic. The CD14 genotype was determined in 228 patients with brucellosis from a rural area and 129 healthy volunteers from the same area. The prevalence of genotype TT was significantly higher in the patients while the controls showed higher prevalence of genotype CC (34.5% vs 15.5%, 15.4% vs 25.6%, P = 0.009). Multiple logistic regression analysis after adjustment for gender demonstrated that the patients who were homozygous for allele T of promoter of CD14 gene had a significantly higher risk for developing brucellosis with odds ratio of 3.03 (95% CI, 5.2, 1.75 P = 0.0004). The existence of homozygous genotype of allele T of CD14 was an independent determinant for occurrence of arthritis among the patients with brucellosis (odds ratio of 3.92 (95% CI, 2.93, 5.88, P = 0.001).Our findings provide suggestive evidence of association of the CD14 gene polymorphism with susceptibility to development of brucellosis in Iranian populations. PMID- 16698682 TI - Cytotoxic lymphocytes for the clinic. PMID- 16698683 TI - AML-loaded DC generate Th1-type cellular immune responses in vitro. AB - BACKGROUND: The generation of AML-specific T-lymphocyte responses by leukemia derived DC has been documented by multiple investigators and is being pursued clinically. An obstacle to widespread use of this strategy is that it has not been possible to generate leukemic DC from all patients, and an alternative approach is needed if the majority of leukemia patients are to receive therapeutic vaccination in conjunction with other treatment protocols. METHODS: In the present study, we generated DC from CD14-selected monocytes isolated from healthy donor PBPC and loaded them with a total cell lysate from AML patient blasts. RESULTS: Immature in vitro-derived DC exhibited robust phagocytic activity, and mature DC demonstrated high expression of CD80, CD83, CD86 and the chemokine receptor CCR7, important for DC migration to local lymph nodes. Mature, Ag-loaded DC were used as APC for leukemia-specific cytotoxic T-lymphocyte (CTL) induction and demonstrated cytotoxic activity against leukemic targets. CTL lysis was Ag-specific, with killing of both allogeneic leukemic blasts and autologous DC loaded with allogeneic AML lysate. HLA-matched controls were not lysed in our system. DISCUSSION: These data support further research into the use of this strategy as an alternative approach to leukemia-derived DC vaccination. PMID- 16698684 TI - Manufacturing of gene-modified cytotoxic T lymphocytes for autologous cellular therapy for lymphoma. AB - BACKGROUND: The production of therapeutic T-cell populations for adoptive immunotherapy of cancer requires extensive ex vivo cell processing, including the isolation or creation of Ag-specific T cells and their subsequent propagation to clinically relevant numbers. These procedures must be performed according to the principles of current good manufacturing practices (cGMP) for phase I clinical trials to ensure the identity, purity potency and safety of the cellular product. In this report we describe our approach to manufacturing and characterizing bulk populations of gene-modified autologous T cells for use in treating follicular lymphoma. METHODS: PBMC from healthy donors, obtained after informed consent, were stimulated in vitro with Ab to CD3epsilon (OKT3) and recombinant human IL-2 and then electroporated with plasmid DNA containing a human CD19-specific chimeric Ag receptor (CAR) gene and HSV-1 thymidine kinase (TK) gene. Stably transfected cells were selected in cytocidal concentrations of hygromycin B over multiple 14-day stimulation culture cycles and then cryopreserved. Vials of cryopreserved/selected T cells were used to initiate T-cell expansion cultures to produce cell products for clinical infusion. These cultures were characterized for phenotype, function and suitability for use in adoptive immunotherapy studies. RESULTS: Our results demonstrate that bulk populations of gene-modified T cells derived from peripheral blood of healthy donors express CD19+ chimeric Ag receptor at low levels and can specifically lyse CD19+ target cells in vitro. These cells display a differentiated T-effector phenotype, are sensitive to ganciclovir-mediated killing and display a non-transformed phenotype. TCR Vbeta usage indicated that all populations tested were polyclonal. Ex vivo cell expansion from cryopreserved cell banks is sufficient to produce doses of between 5 x 10(9) and 1 x 10(10) cells/run. One of three transductions resulted in a population of cells that was not suitable for infusion but was identified during release testing. No populations displayed any evidence of bacterial, fungal or mycoplasma contamination. DISCUSSION: We have established a manufacturing strategy that is being used to produce T cells for a phase I clinical trial for follicular lymphoma. Genetically modified T cells have been characterized by cell surface marker phenotype, functional activity against CD19+ targets and requisite safety testing. These pre-clinical data confirm the feasibility of this approach to manufacturing T-cell products. PMID- 16698685 TI - Type I IFN-mediated enhancement of anti-leukemic cytotoxicity of gammadelta T cells expanded from peripheral blood cells by stimulation with zoledronate. AB - BACKGROUND: In order to establish efficient gammadelta T-cell based tumor immunotherapy, we explored a method to enhance the cytotoxicity of gammadelta T cells against leukemia cells by stimulating gammadelta T cells with type I IFN. METHODS: Gammadelta T cells were expanded from normal PBMC by culturing with zoledronate and a low concentration of IL-2 for 2 weeks. For the activation of gammadelta T cells, gammadelta T cells were cultured with type I IFN (HLBI, IFN alpha2b and IFN-beta) for 1-3 days. The cytotoxicity of HLBI-activated gammadelta T cells against leukemia cell lines and fresh leukemia cells was evaluated by 51Cr-release assay. RESULTS: Gammadelta T cells, which were expanded and purified with magnetic beads using an anti-gammadelta TCR MAb, were demonstrated to be cytotoxic against leukemia cell lines of both lymphoid and myeloid origin and fresh myeloid leukemia cells. By culturing expanded gammadelta T cells with type I IFN, the expression of the activation marker CD69 was increased and the cytometric bead array showed an elevated production of IFN-gamma by gammadelta T cells. In addition, the cytotoxicity of gammadelta T cells against leukemia cells was definitely enhanced by culturing gammadelta T cells with HLBI. DISCUSSION: The present study has demonstrated that type I IFN could enhance the anti leukemic cytotoxicity of expanded gammadelta T cells, which implies that in vitro bisphosphonate (such as zoledronate)-expanded and type I IFN-activated gammadelta T cells could be applied to immunotherapy for hematologic malignancies such as leukemia and lymphoma. PMID- 16698686 TI - CD34+-derived CD11c+ + + BDCA-1+ + CD123+ + DC: expansion of a phenotypically undescribed myeloid DC1 population for use in adoptive immunotherapy. AB - BACKGROUND: DC are commonly defined as HLA-DR+/Lin- cells that can be CD11c+ + + CD123+/ -, termed DC1/myeloid DC that induce a Th1 response, or CD11c- CD123+ + +, termed DC2/lymphoid DC that induce a Th2 response. However, significant heterogeneity within DC preparations is apparent and supports the existence of several distinct DC subpopulations. This study aimed to expand and characterize CD34+ DC for use in immunotherapy. METHODS: CD34+ cells were seeded at 1 x 10(5)/mL and expanded for 14 days in RPMI + 10% autologous plasma supplemented with GM-CSF, IL-4, Flt-3L and SCF. Maturation was induced with TNF-alpha and PGE2 for 2 days. DC were analyzed morphologically, phenotypically with a panel of MAb to lineage and DC markers, and functionally in MLR, T-cell assays and T-cell cytokine secretion by ELISA. RESULTS: Significant cellular expansion was observed: 60+/-5 x 10(6) DC from 1 x 10(6) CD34+ cells (n=28). Phenotypically DC were characterized as HLA-DR+ +, CD11c+ + +, CD80+ +, CD83+, CD86+ +, CD123+ +, CD15+ +, CD33+ +, BDCA-1+ +, CD4+ and Lin-. DC displayed potent allostimulatory capacity and efficient presentation of KLH and tetanus toxin. DC-primed T cells secreted IFN-gamma (Th1); however, no detectable IL-4 (Th2) was noted. DISCUSSION: We present features of CD34+ DC that have not been previously described. The CD34+ DC generated represent a population of myeloid DC functioning as DC1 but phenotypically expressing markers characteristic of both DC1 and DC2. This novel DC population is capable of inducing naive T-cell responses and can be expanded to clinically useful numbers. CD34+-derived DC represent attractive candidates for use in adoptive T-cell immunotherapy. PMID- 16698687 TI - Ex vivo expansion of non-MHC-restricted cytotoxic effector cells as adoptive immunotherapy for myeloma. AB - BACKGROUND: PBMC can be expanded ex vivo into aggressive cytotoxic effector cells (CEC) comprising T, NK and NKT cells. We identified the phenotype, cytotoxicity and mechanisms of killing of these CEC. METHODS: CY- and G-CSF-mobilized PBMC from myeloma patients were placed in Aim-V serum-free medium, IL-2 (50 IU/mL) and OKT-3 (50 ng/mL). Cytotoxicity was evaluated by selectively blocking the TCR, MHC class I or NKG2D receptor. RESULTS: The CEC expanded three-fold by day 7 and aggressively lysed myeloma cells (41.9%) compared with day 0 (4%; P=0.012). CD8+ CD56+ NKT cells performed the majority of lysis. The CD8+ cells greatly increased NKG2D expression during culture (P=0.005). Cytotoxicity correlated with target NKG2D ligand expression (P=0.0002). Blocking the TCR or MHC class I did not affect cytotoxicity (P>0.22). CD8+ cell-mediated lysis dropped 48% when the NKG2D receptor was blocked. Day 7 CEC aggressively lysed myeloma cells in an MHC- and non-MHC-restricted fashion, through the NKG2D receptor. DISCUSSION: Because MHC expression is often down-regulated on tumor cells and the NKG2D ligands are generally specific to malignant cells, the adoptive transfer of CEC that kill through different pathways may circumvent tumor-resistant mechanisms and improve outcomes. PMID- 16698688 TI - Ex vivo expanded umbilical cord blood T cells maintain naive phenotype and TCR diversity. AB - BACKGROUND: Umbilical cord blood (CB) is a promising source of hematopoietic stem cells for allogeneic transplantation. However, delayed engraftment and impaired immune reconstitution remain major limitations. Enrichment of donor grafts with CB T cells expanded ex vivo might facilitate improved T-cell immune reconstitution post-transplant. We hypothesized that CB T cells could be expanded using paramagnetic microbeads covalently linked to anti-CD3 and anti-CD28 Ab. METHODS: CB units were divided into three fractions: (1) cells cultured without beads, (2) cells cultured with beads and (3) cells cultured with beads following CD3+ magnetic enrichment. All fractions were cultured for 14 days in the presence of IL-2 (200 IU/mL). RESULTS: A mean 100-fold expansion (range 49-154) of total nucleated cells was observed in the CD3+ magnetically enriched fraction. Following expansion, CB T cells retained a naive and/or central memory phenotype and contained a polyclonal TCR diversity demonstrated by spectratyping. DISCUSSION: Our data provide evidence that naive and diverse CB T cells may be expanded ex vivo and warrant additional studies in the setting of human CB transplantation. PMID- 16698689 TI - Fresh PBSC harvests, but not BM, show temperature-related loss of CD34 viability during storage and transport. AB - BACKGROUND: The optimum conditions for storage and transport of freshly harvested HPC in the liquid state are uncertain. It is not specified in commonly applied standards for stem cell transplantation. We used a viable CD34 assay to determine the optimum temperature for maintaining progenitor cell viability in freshly harvested BM and PBSC. Our aim was to identify standardized conditions for storage and transport of marrow or peripheral blood products that would optimize CD34 recovery, leading to better transplant outcomes. METHODS: Samples were aseptically removed from 46 fresh HPC harvests (34 PBSC and 12 BM) and stored at refrigerated temperature (2-8 degrees C), room temperature (18-24 degrees C) and 37 degrees C for up to 72 h. Samples were analyzed for viable CD34+ cells/microL at 0, 24, 48 and 72 h. RESULTS: The mean viable CD34+ yield prior to storage was 7.7 x 10(6)/kg (range 0.7-30.3). The mean loss of viable CD34+ cells in HPC products at refrigerated temperature was 9.4%, 19.4% and 28% at 24, 48 and 72 h, respectively. In contrast, the mean loss of viable CD34+ cells at room temperature was 21.9%, 30.7% and 43.3% at 24, 48 and 72 h, respectively. No viable CD34+ cells remained after storage at 37 degrees C for 24 h. Only PBSC products and not BM showed temperature-related loss of CD34 viability. Greater loss of viable CD34+ cells was observed for allogeneic PBSC compared with autologous PBSC. DISCUSSION: These results demonstrate that the optimum temperature for maintaining the viability of CD34+ cells, during overnight storage and transport of freshly harvested HPC, is 2-8 degrees C. These findings will allow the development of standard guidelines for HPC storage and transport. PMID- 16698690 TI - Adipose tissue-derived mesenchymal stem cell yield and growth characteristics are affected by the tissue-harvesting procedure. AB - BACKGROUND: Adipose tissue contains a stromal vascular fraction that can be easily isolated and provides a rich source of adipose tissue-derived mesenchymal stem cells (ASC). These ASC are a potential source of cells for tissue engineering. We studied whether the yield and growth characteristics of ASC were affected by the type of surgical procedure used for adipose tissue harvesting, i.e. resection, tumescent liposuction and ultrasound-assisted liposuction. METHODS: Frequencies of ASC in the stromal vascular fraction were assessed in limiting dilution assays. The phenotypical marker profile of ASC was determined, using flow cytometry, and growth kinetics were investigated in culture. ASC were cultured under chondrogenic and osteogenic conditions to confirm their differentiation potential. RESULTS: The number of viable cells in the stromal vascular fraction was affected by neither the type of surgical procedure nor the anatomical site of the body from where the adipose tissue was harvested. After all three surgical procedures, cultured ASC did express a CD34+ CD31- CD105+ CD166+ CD45- CD90+ ASC phenotype. However, ultrasound-assisted liposuction resulted in a lower frequency of proliferating ASC, as well as a longer population doubling time of ASC, compared with resection. ASC demonstrated chondrogenic and osteogenic differentiation potential. DISCUSSION: We conclude that yield and growth characteristics of ASC are affected by the type of surgical procedure used for adipose tissue harvesting. Resection and tumescent liposuction seem to be preferable above ultrasound-assisted liposuction for tissue engineering purposes. PMID- 16698691 TI - Generation of a bioartificial fibromuscular tissue with autoregenerative capacities for surgical reconstruction. AB - INTRODUCTION: Anecdotal clinical reports denote first tissue engineering applications entering medical practice. Currently it is still unknown, if these new types of implants will tolerate the specific needs in cancer patients undergoing postoperative chemo- and radiotherapy. METHODS: We implemented a radiotherapy protocol (cumulative dosis 40 Gy) on generated human bioartificial fibromuscular tissues in vitro. We monitored tissue vitality during radiotherapy and tissue recovery (8 weeks follow up period) applying histological methods. RESULTS: The biopsy procedure and seeding techniques yielded a viable 3 dimensional bioartificial human tissue. Radiation resulted in immediate devitalization without destroying tissue integrity. The bioartificial tissue recovered entirely in vitro within 6 weeks. CONCLUSION: Bioartificial human implants appear applicable for surgical reconstruction in oncologic patients potentially facing postoperative radiotherapy. PMID- 16698693 TI - Brain-derived neurotrophic factor treatment does not improve functional recovery after hair cell regeneration in the pigeon. AB - CONCLUSIONS: Brain-derived neurotrophic factor (BDNF) supply to the inner ear does not improve the time course or the extent of functional recovery after hair cell regeneration. Specifically it does not improve the residual threshold elevation observed after the completion of spontaneous recovery. OBJECTIVE: The avian inner ear is capable of hair cell regeneration and substantial functional recovery, but residual hearing deficits remain. We investigated whether functional recovery can be improved by intracochlear application of BDNF, which plays an important role in auditory ontogenesis and maintenance during adult life. METHODS: Hair cells in adult pigeons were destroyed by local application of gentamicin. After 3 days either BDNF or control solution was administered to the scala tympani by implanted osmotic minipumps for 8 weeks. Auditory brain stem responses (ABR) to tone pips were used to assess recovery of hearing thresholds in both groups. RESULTS: The application of gentamicin caused a frequency dependent hearing loss that ranged from 24.8 dB SPL at low frequencies to 66.2 dB SPL at high frequencies. After day 10 substantial recovery was observed, but a significant threshold shift remained. The time course of recovery in the control and BDNF-treated groups was similar, without significant residual threshold differences in any frequency range. PMID- 16698694 TI - Inner ear and facial nerve complications of acute otitis media with focus on bacteriology and virology. AB - CONCLUSION: Among 20 patients with inner ear complications and/or peripheral facial palsy secondary to acute otitis media (AOM) a proven or probable bacteriological cause was found in 13 (65%). In seven patients (35%), a proven or probable viral cause was found. Only two of the patients (10%), with a proven bacterial AOM and a clinical picture of a purulent labyrinthitis in both, together with a facial palsy in one, had a substantial degree of dysfunction. Although the number of patients in this study is relatively low our findings show that inner ear complications and facial palsy due to AOM can be of both bacterial and viral origin. Severe sequelae were found only where a bacterial origin was proven. OBJECTIVES: Inner ear complications and/or peripheral facial palsy secondary to AOM are rare. The general understanding is that they are due to bacterial infections. However, in some of these patients there are no clinical or laboratory signs of bacterial infections and they have negative bacterial cultures. During recent years different viruses have been isolated from the middle ear or serologically proven in AOM patients and are thought to play a pathogenetic role. We suggest that in some cases of AOM complications from the inner ear and the facial nerve can be caused by viruses. The purpose of our study was to analyze infectious agents present in patients with inner ear complications and/or facial palsy arising from AOM. PATIENTS AND METHODS: The medical records of 20 patients who had inner ear complications and/or facial palsy following AOM ( unilateral in 18, bilateral in 2) between January 1989 and March 2003 were evaluated. Bacterial cultures were carried out for all patients. Sera from 12 of the patients were stored and tested for a battery of specific viral antibodies. In three patients, investigated between November 2002 and March 2003, viral cultures were also performed on samples from the middle ear and nasopharynx. RESULTS: Nineteen patients had inner ear symptoms. Eight of them had a unilateral sensorineural hearing loss and vertigo, three had vertigo as an isolated symptom and one, with bilateral AOM, had bilateral sensorineural hearing loss. Seven patients had a combination of facial palsy and inner ear symptoms (unilateral sensorineural hearing loss in three, unilateral sensorineural hearing loss and vertigo in two, bilateral sensorineural hearing loss and vertigo in one, with bilateral AOM, and vertigo alone in one). One patient had an isolated facial palsy. Healing was complete in 11 of the 20 patients. In seven patients a minor defect remained at follow-up (a sensorineural hearing loss at higher frequencies in all). Only two patients had obvious defects (a pronounced hearing loss in combination with a moderate to severe facial palsy (House-Brackman grade 4) in one, distinct vestibular symptoms and a total caloric loss in combination with a high-frequency loss in the other. Eight patients had positive bacteriological cultures from middle ear contents: Streptococcus pneumoniae in two, beta hemolytic Streptococcus group A in two, beta-hemolytic Streptococcus group A together with Staphylococcus aureus in one, Staph. aureus alone in one and coagulase-negative staphylococci (interpreted as pathogens) in two. In the 12 patients with negative cultures, there was a probable bacteriological cause due to the outcome in SR/CRP and leukocyte count in five. In four patients serological testing showed a concomitant viral infection that was interpreted to be the cause (varicella zoster virus in two, herpes simplex virus in one and adenovirus in one.) In three there was a probable viral cause despite negative viral antibody test due to normal outcome in SR/CRP, normal leukocyte count, serous fluid at myringotomy and a relatively short pre-complication antibiotic treatment period. PMID- 16698695 TI - High resolution micro-CT scanning as an innovative tool for evaluation of the surgical positioning of cochlear implant electrodes. AB - X-ray microtomography (micro-CT) is a new technique allowing for visualization of the internal structure of opaque specimens with a quasi-histological quality. Among multiple potential applications, the use of this technique in otology is very promising. Micro-CT appears to be ideally suited for in vitro visualization of the inner ear tissues as well as for evaluation of the electrode damage and/or surgical insertion trauma during implantation of the cochlear implant electrodes. This technique can greatly aid in design and development of new cochlear implant electrodes and is applicable for temporal bone studies. The main advantage of micro-CT is the practically artefact-free preparation of the samples and the possibility of evaluation of the interesting parameters along the whole insertion depth of the electrode. This paper presents the results of the first application of micro-CT for visualization of the inner ear structures in human temporal bones and for evaluation of the surgical positioning of the cochlear implant electrodes relative to the intracochlear soft tissues. PMID- 16698696 TI - Combining perimodiolar electrode placement and atraumatic insertion properties in cochlear implantation -- fact or fantasy? AB - CONCLUSIONS: Except for basal cochlear traumatization, all specimens implanted into scala tympani showed atraumatic insertion properties and good perimodiolar electrode positioning. Cochleostomy preparation and placement can have a significant impact on levels of basal cochlear trauma. OBJECTIVE: In the past, perimodiolar cochlear implant electrodes increased the risk for intracochlear traumatization when compared to free-fitting arrays. Recently, however, clinical evidence for atraumatic perimodiolar implantations with preservation of residual hearing has been described. The aim of this paper was to histologically evaluate a perimodiolar cochlear implant array for its insertion properties in cadaver human temporal bones. Surgical and electrode factors, as well as preparation artifacts influencing intracochlear trauma, were considered in the evaluation. MATERIALS AND METHODS: Sixteen human temporal bones were harvested up to 24 hours post mortem and implanted immediately with the Nucleus 24 Contour Advance cochlear implant electrode array. Implantations were either performed using a regular caudal approach cochleostomy or through the round window membrane. After implantation, all bones underwent special histological processing, which allowed sectioning of undecalcified bone. Insertion properties were evaluated according to a grading system. RESULTS: Fourteen specimens were implanted into scala tympani and only two exhibited basal trauma attributable to electrode insertion characteristics. Two bones were implanted into scala vestibuli after causing trauma in the region of the cochleostomy. Insertion depths ranged from 180 degrees to 400 degrees. All bones showed good perimodiolar electrode positioning. Basal trauma due to surgical issues and histological artifacts was present in 10 of 16 bones. PMID- 16698697 TI - Prevalence of upper and lower airway symptoms: the Skovde population-based study. AB - CONCLUSION: This study was able to present representative data from Sweden as a whole and illuminates the epidemiological background of symptoms of the upper and lower airways. Current regular smokers, compared with non-smokers, had more problems with nasal blockage, cough and asthma. The relationship between the upper and lower airways was confirmed as regards airway symptoms. BACKGROUND: Epidemiology of common respiratory symptoms is basic information in health care. This study has focused on the prevalence of upper and lower airway symptoms. PATIENTS AND METHODS: A random sample of 1900 age- and gender-stratified adults was recruited from a general population for this study, which included a structured interview about symptoms of nasal secretions and blockage, cough and asthma, and a clinical examination with nasal endoscopy, and peak nasal inspiratory flow (PNIF). Smoking habits were evaluated by a questionnaire for current and previous smoking. RESULTS: A response rate of 73% was achieved. The prevalence of various symptoms varied between age strata and gender. There was a statistically significant relationship between subjectively experienced nasal blockage and objectively observed nasal congestion. PNIF was negatively related to age and height and was increased in male gender. Smoking was most common among females and related to nasal blockage, cough and asthma, but not to nasal secretions. Significant relationships between the upper and lower airways were found. PMID- 16698698 TI - Bacteriological findings and antimicrobial susceptibility in chronic sinusitis with nasal polyp. AB - CONCLUSIONS: We recommend amoxacillin/clavulanate, cephalosporins and macrolides rather than penicillin as the first-line drug in chronic sinusitis with nasal polyps. In cases where there is no improvement of symptoms, cultures should be taken from the middle meatus, followed by appropriate selection of second-line antibiotics according to the sensitivity test results. OBJECTIVE: To investigate the causative bacteria and the antimicrobial susceptibility in patients with chronic sinusitis and nasal polyps in Korea. MATERIALS AND METHODS: The bacteriology and antimicrobial susceptibility of maxillary sinus aspirates from 81 patients were evaluated. RESULTS: Aerobes were isolated from 58.0% of the cultures from the middle meatus and from 48.1% of those from the maxillary sinus. Staphylococcus aureus, Haemophilus influenzae, and Streptococcus pneumoniae were the most prevalent aerobic pathogens. Anaerobes were isolated from 8.6% of the cultures from the middle meatus and from 18.5% of the cultures from the maxillary sinus. The predominant anaerobic organisms were Prevotella and Peptostreptococcus in adults but none of them were cultured in children. A high rate of concordance of the middle meatus and maxillary sinus was noted. Monomicrobial infection was most commonly observed. Ampicillin-resistant H. influenzae isolates were cultured in 46% of the cases. Penicillin resistance rates were 93% for Staph. aureus; 25% of Strep. pneumoniae were intermediate and 25% were resistant. PMID- 16698699 TI - Expression of Notch1 and Hes5 in the developing olfactory epithelium. AB - CONCLUSION: Notch signaling pathway may play an important role in the development of the olfactory epithelium (OE). Objectives. To elucidate whether the Notch signaling pathway mediates the developmental processes in the developing OE. MATERIALS AND METHODS: The expression of Notch1 and Hes5 in the developing OE of mice with ages ranging from embryonic day (E) 11 to postnatal day (PN) 14 was examined. RESULTS: As detected by in situ hybridization, Notch1 was expressed in scattered cells located in the basal portion of the embryonic OE and later in the cell layer adjacent to the basal lamina from E11 to PN14. Hes5 was expressed in scattered cells located in the basal portion of the embryonic OE from E11. However, at the late embryonic stage, the number of Hes5-positive cells decreased and after birth distinct Hes5-positive cells were not observed in the OE. PMID- 16698700 TI - Ear, nose and throat manifestations of Churg-Strauss syndrome. AB - CONCLUSION: Ear, nose and throat (ENT) involvement is common in Churg-Strauss syndrome (CSS), usually manifesting as allergic rhinitis and chronic rhinosinusitis with or without polyps. Otolaryngologists may play a pivotal role in making an early diagnosis of this disease. OBJECTIVES: CSS is a systemic vasculitic disorder that affects small to medium-sized blood vessels. Although the cause of CSS remains unknown, tissue damage seems more likely to be mediated by activated eosinophils. Patients affected by CSS frequently have ENT manifestations, which are often present at the time of disease onset and may represent relevant clues for the diagnosis. Thus, our objective was to present the ENT manifestations at the onset, at the diagnosis and at some point during the course of the disease in a series of patients with CSS collected at a single center. MATERIALS AND METHODS: Twenty-eight patients with CSS, as defined according to the 1990 American College of Rheumatology classification criteria, were identified. Twenty-one (75%) of these patients had ENT involvement. We evaluated the clinical course, laboratory data, histologic findings, treatment and outcomes. RESULTS: Of the 21 patients, 13 (61.9%) had ENT involvement at asthma onset and 8 (38%) at diagnosis or during follow-up. The most common ENT manifestations were allergic rhinitis in 9 (42.8%) patients and nasal polyposis in 16 (76.1%). Three (14.2%) patients developed chronic rhinosinusitis without polyps, three (14.2%) had nasal crusting, one (4.7%) serous otitis media, one (4.7%) purulent otitis media, two (9.5%) progressive sensorineural hearing loss, and one (4.7%) unilateral facial palsy. Corticosteroid therapy associated with immunosuppressive drugs usually yielded improvement or stabilization. PMID- 16698701 TI - Treatment of branchial cleft cyst with intracystic injection of OK-432. AB - CONCLUSIONS: The data suggest that branchial cleft cyst (BCC) can be primarily treated with OK-432 sclerotherapy and only the remaining lesions with excision. OBJECTIVES: To evaluate the effectiveness of sclerotherapy using OK-432 in the treatment of BCC. MATERIALS AND METHODS: We treated 12 BCC patients (3 males and 9 females; mean age 25 years) with OK-432 sclerotherapy at an outpatient clinic. The cystic fluids were aspirated and diagnosed by cytomorphology and DNA cytometric analysis to exclude malignancy. The fluid aspirated from the cyst was replaced with an equal volume of OK-432 solution. The sizes of cysts were measured and compared before and after injection. The remaining cysts were excised and histopathologically compared with the excised BCCs that had not been treated with OK-432. RESULTS: Seven of 12 patients (58%) showed a complete response after OK-432 injection, administered one to three times. Three patients (25%) had only partial response and two (17%) were stationary. Five patients with remaining lesions underwent excision. There was no difficulty in dissecting around the cysts and no increased morbidity during operation. None of the patients had evidence of recurrences or malignancies developing during the follow up period (mean 21 months, range 17-26 months). There were no major side effects except fever after sclerotherapy. PMID- 16698702 TI - Three-dimensional laryngeal model for planning of laryngeal framework surgery. AB - CONCLUSION: The three-dimensional prototype model was useful for planning of laryngeal framework surgery. OBJECTIVE: To discuss the usefulness of a three dimensional laryngeal model for laryngeal framework surgery. MATERIALS AND METHODS: A three-dimensional laryngeal model was created based on the postoperative helical computed tomography (CT) data of the larynx (case 1) which underwent lateral cricoarytenoid muscle (LCA) pull surgery. LCA pull surgery is a kind of arytenoid adduction for unilateral vocal cord paralysis. A three dimensional model of case 1 larynx was prototyped using a selective laser sintering method. In case 1, the patient's voice did not improve after LCA pull surgery. The three-dimensional model revealed that the original surgical procedure was not appropriate to obtain optimal arytenoid adduction. According to the analysis of this three-dimensional model, we changed the surgical approach and performed this new refined LCA pull surgery on another patient with unilateral vocal cord paralysis (case 2). RESULTS: We were able to pull LCA precisely in case 2. Three-dimensional CT of case 2 after refined LCA pull surgery allowed the correct pulling of LCA and complete adduction of arytenoid. The postoperative voice improved remarkably. PMID- 16698703 TI - High-speed digital imaging and electroglottography of tracheoesophageal phonation by Amatsu's method. AB - BACKGROUND: Our previous findings have indicated that the thyropharyngeal muscles form a retropharyngeal prominence during alaryngeal phonation via the TE fistula. This prominence forms a so-called 'neoglottis', which is thought to function as the vibratory source. To better understand the mechanism of TE phonation, we analyzed the vibration of the neoglottis using electroglottography (EGG) and a high-speed digital imaging system. PATIENTS AND METHODS: Two volunteers who use TE phonation for their daily speech communication were subjected to this study. The vibrations of the neoglottis were recorded simultaneously as EGG and high speed imaging with acoustic signals. RESULTS: The vibrations of the neoglottis, recorded by means of high-speed digital imaging, were exactly synchronized with the waveforms of the acoustic signals and EGG. CONCLUSIONS: These results further confirm the neoglottis as the source of vibration during tracheoesophageal (TE) phonation. PMID- 16698704 TI - A modified technique for bypass of the external carotid artery to the proximal posterior cerebral artery: an anatomical and technical study. AB - CONCLUSIONS: Our results support the proposition that bypass between the external carotid artery (ECA) and proximal posterior cerebral artery (PCA) can be achieved by using a short saphenous venous graft. The diameters of the ECA and vein graft may provide an increased blood flow with a straighter course. This technique may be helpful for management of patients with vertebrobasilar insufficiency or those requiring a high volume blood flow to the posterior circulation. OBJECTIVES: We aimed to describe a modified technique using a short saphenous vein graft for bypass between the ECA and the PCA in order to use a small length of graft material and increase the patency of the anastomosis. MATERIALS AND METHODS: Ten sides of five cadavers were dissected bilaterally. After a frontotemporal craniotomy and zygomatic arch osteotomy, the middle cranial fossa was exposed. A hole located approximately 2-3 cm posterolateral to the foramen rotunda was created extradurally. The sylvian fissure and the interpeduncular and ambient cisterns were opened. The proximal P2 segment of the PCA was identified. The ECA was found through a cervical incision. A short interposition saphenous vein graft was conducted to pass just behind the ramus mandible to the infratemporal fossa. The bypass between the ECA and P2 segment of the PCA was performed by using a short saphenous vein graft. The diameters of the ECA, P2 segment of PCA and both ends of the saphenous vein graft and its length were measured using an electronic micrometer. RESULTS: The mean cross-clamping time of the PCA was 10.4+/-1.8 min. The mean diameters of the P2 segment of the PCA and ECA were 2.2+/-0.15 mm and 3.83+/-0.28 mm, respectively. The mean length of the saphenous vein graft was 88.8+/-3.8 mm. PMID- 16698705 TI - Complications after CO(2) laser surgery of laryngeal cancer in the elderly. AB - CONCLUSION: Cancer of the larynx in elderly patients should be treated with curative intention, if the extension of the primary tumour allows safe resection. Transoral CO(2) laser microsurgery is associated with a low rate of major complications and can be regarded as suitable even for elderly patients. Age itself should never be the sole factor in deciding which therapy should be undertaken. OBJECTIVES: In the industrialized nations the age group beyond 75 years will grow steadily, requiring special attention by medical professionals in the future. Nowadays laryngeal cancer is often treated by transoral CO(2) laser microsurgery. Because of a variety of comorbidities, the incidence of perioperative complications in the group of elderly patients is of increasing interest. To date, no references in the literature have discussed complications after transoral CO(2) laser microsurgery in this age group. Patients and methods. Twenty-four patients over the age of 75 suffering from squamous cell carcinoma of the larynx and treated by transoral CO(2) laser microsurgery since 1998 were analysed for intraoperative and postoperative complications. RESULTS: There was no evidence of surgery-related complications at all. Postoperatively, 6 of 24 patients complained about dysphagia. Due to intraoperative placement of nasogastric feeding tubes in these six cases further complications such as pneumonia resulting from aspiration could have been avoided. PMID- 16698706 TI - Diagnostic work-up and outcome of cervical metastases from an unknown primary. AB - CONCLUSIONS: An intensive diagnostic work-up including (18)F-fluorodeoxyglucose positron emission tomography (FDG-PET) detects many unknown primary tumours, leads to a low emergence rate of primary tumours, and selects carcinoma of unknown primary with much more favourable results after neck dissection and postoperative radiotherapy. OBJECTIVE: To investigate the optimal diagnostic approach and best treatment modality for rare head and neck cancer of unknown primary. PATIENTS AND METHODS: In a retrospective study, 69 patients admitted from 1987 to 2002 with cervical lymph node metastases without apparent primary were reviewed. Test characteristics of all diagnostic procedures were calculated. Disease-free and overall survival rates were calculated. Major prognostic factors were analysed uni-variously. RESULTS: At the primary site FDG-PET showed the best sensitivity with 69% and the highest negative predictive value with 87%. Computed tomography and magnetic resonance imaging had a better specificity with 87% and 95%, respectively. The primary tumour was detected in 23 cases (33%). Frequent primary tumour origin was the palatine tonsil (n=8, 35%), base of the tongue (n=6, 26%) and lung (n=4, 17%). All patients with unknown primary were treated by neck dissection. Adjuvant radiotherapy was performed in 26 patients (57%), concurrent radiochemotherapy was performed in 12 patients (26%). The primary emergence rate was 7%. The 5-year overall survival rate was inferior in patients with detected primary in comparison with patients with unknown primary (22% versus 52%). Significant prognostic factors in case of unknown primary were M stage, smoking, alcohol consumption and tonsillectomy. Radiotherapy but not chemotherapy with carboplatin influenced the overall survival. PMID- 16698707 TI - Recombinant erythropoietin beta enhances growth of xenografted human squamous cell carcinoma of the head and neck after surgical trauma. AB - CONCLUSION: Treatment of anaemia with recombinant human erythropoietin beta (rHuEpo) in patients with squamous cell carcinoma of the head and neck (HNSCC) undergoing curative radiotherapy does not improve cancer control. In fact, incompletely resected patients with HNSCC receiving radiation in combination with rHuEpo showed poorer loco-regional progression-free survival than patients receiving radiation in combination with placebo. It could be hypothesized that the effects of rHuEpo on tumour cell growth might only be manifested in vivo and after cell trauma, and that treating anaemia with rHuEpo might contribute to poor outcome after incomplete surgical resection. OBJECTIVE: The aim of the present study was to examine the effect of rHuEpo alone and in combination with surgical trauma on the growth of human squamous cell carcinoma in vivo, xenografted onto nude mice. MATERIALS AND METHODS: The surgical trauma was inflicted through subcutaneous transection of the tumour with a needle. Immunohistochemical staining verified expression of the EPO receptor in tumour cells. RESULTS: rHuEpo alone had no effect on the growth of xenografted HNSCC. However, a significant increase in tumour growth was observed after surgical trauma in combination with rHuEpo compared with surgery alone (p = 0.0008). PMID- 16698708 TI - Endolymphatic hydrops as a cause of audio-vestibular manifestations in relapsing polychondritis. AB - Relapsing polychondritis (RP) is characterized by inflammation and subsequent degeneration of cartilage. We report a 61-year-old woman who had RP with audio vestibular manifestations. She was also diagnosed as having a myelofibrosis with myeloid metaplasia (MMM). Bilateral endolymphatic hydrops (EH) was confirmed by dominant -SP/AP of the electrocochleogram (ECochG). When thalidomide and prednisolone were prescribed for the treatment of MMM, symptoms of RP -- including the inner ear dysfunction -- were ameliorated. Isosorbide, one of the osmotic diuretics commonly used for the treatment of Meniere's disease (MD) in Japan, was also effective in keeping her free from inner ear dysfunction. This is the first report to confirm the existence of EH in a patient with RP with audio vestibular manifestations. We suppose that an immunological imbalance due to MMM, in conjunction with a specific immunogenetic background, may have played a role in the pathogenesis of RP and the formation of EH in this patient. PMID- 16698709 TI - Cervical bronchogenic cyst: asymptomatic neck mass in an adult male. AB - Bronchogenic cysts are rare congenital malformations of ventral foregut development. They are usually located in the mediastinum and intrapulmonary regions. Localization in the cervical area is unusual and the majority of cases reported have been found in the pediatric population: the literature reports few cases in adults. We describe a 57-year-old male who presented an asymptomatic right lateral neck mass. Diagnostic studies included chest X-ray, pharyngo esophagus double-contrast X-ray, computed tomography (CT), and bronchoscopy. The mass was excised through a transverse right cervical skin incision. The right lateral neck mass of the patient was identified as a bronchogenic cyst. The embryology, the presentation, the pathological and radiological evaluation, treatment of the cyst and a review of the English literature are reported in this paper. Cervical bronchogenic cysts are usually diagnosed in the pediatric population; these lesions are rare in adults. We suggest that the clinical observation of an asymptomatic lateral neck mass in an adult should include the possibility of a bronchogenic cyst in the differential diagnosis. Surgical excision is the elective treatment for this tumor, in order to prevent complications including infection, compression symptoms, malignant transformation, and the rare but fatal air embolism. PMID- 16698710 TI - Carotid artery pseudoaneurysm as a rare sequela of surgery for laryngeal cancer. AB - Formation of carotid artery pseudoaneurysm is uncommon after intensive treatment for head and neck cancer. We encountered a case of postoperative formation of pseudoaneurysm at the left carotid bifurcation in a diabetic man. The risk factors for carotid artery pseudoaneurysm in the patient included diabetes mellitus, previous radiotherapy to the neck, neck dissection, and postoperative Staphylococcus aureus infection secondary to accidental pharyngocutaneous fistula. We successfully obliterated this pseudoaneurysm using a Smart stent and detachable coils without neurological deficits. The possibility of vascular injury after treatment for head and neck cancer must be considered, especially in patients with risk factors for it due to previous treatment. PMID- 16698716 TI - Proposed mode of action for in utero effects of some phthalate esters on the developing male reproductive tract. AB - Phthalate esters containing a straight-chain backbone of 4-6 carbons have demonstrated testicular toxicity and infertility in adult and pre-adolescent rats, mice, hamsters, and ferrets. In recent years, these same phthalates have been shown to interfere with the normal development of the male reproductive tract in rodents and rabbits. The review presented here summarizes studies that provide evidence of a mode of action for these effects. The data indicate that C4 C6 phthalate esters inhibit processes in the Leydig cell, such as the synthesis of testosterone (T) and production of insulin-like factor 3 (insl3), both of which are required for normal development of male genitalia. A proposed secondary effect of reduced androgen production is on Sertoli cells, resulting in failure to proliferate and interference with cell-cell communication (gap-junction intracellular communication) leading to the development of large multinucleate gonocytes. The possibility that phthalates act directly on the Sertoli cells to interfere with intracellular communication is not excluded. The strength, consistency, and plausibility of the proposed mode of action and alternate modes of action are discussed. PMID- 16698717 TI - Ectopic uterine ovarian tissue in cynomolgus monkeys. AB - Spontaneous ectopic uterine ovarian tissue was found in 9 of 118 cynomolgus monkeys (Macaca fascicularis), an incidence of 7.6%. These ectopic tissues were minute and were incidentally observed in the parametrium by microscopic examination. They were composed of primordial or primary follicles and ovarian stroma. Ectopic ovarian tissue may result from disturbed migration of the ovarian primordium during embryogenesis. PMID- 16698718 TI - Species and sex differences in susceptibility to olfactory lesions among the mouse, rat and monkey following an intravenous injection of vincristine sulphate. AB - Species and sex differences in susceptibility to vincristine sulphate (VCR) induced olfactory epithelial lesions were investigated among the BALB/c mice, Crj: CD(SD) IGS rats and common marmoset monkeys following a single intravenous administration on day 1. As dosage levels, the 0.17-fold LD10, 0.6-fold LD10 and LD10 were used for mice and rats, and a maximum tolerated dose (MTD) was chosen only for monkeys. The order of strength of VCR action on peripheral neuropathic signs, body weight gain, and hematological parameters was mice > rats > monkeys, without clear sex differences. Histopathologically, on day 2, single cell death in the olfactory epithelium and vomeronasal organ was observed only in male mice at LD10, and in female mice at 0.6-fold LD10 or more. On day 5, the olfactory epithelium in these mice showed regenerative proliferation suggesting a sign of recovery. On day 10, axonopathy and demyelination in the sciatic and trigeminal nerves were noted in mice of both sexes at 0.6-fold LD10 or more. In rats and monkeys of either sex, however, no morphological changes were observed at any dose level. In conclusion, mice, particularly females, were shown to be more susceptible to VCR-induced apoptosis in the olfactory epithelium than rats and monkeys. PMID- 16698719 TI - Existence of no hepatocarcinogenic effect levels of 2-amino-3,8 dimethylimidazo[4,5-f]quinoxaline with or without coadministration with ethanol. AB - There is increasing evidence of the existence of no effect levels for genotoxic carcinogens. However, only limited information is available regarding dose response curves for combination effects of multiple carcinogens at low dose. In the present study, 280 male F344 rats were divided into 14 groups to determine the effects of co-administration of various doses of 2-amino-3,8 dimethylimidazo[4,5-f]quinoxaline (MeIQx) and 10% ethanol on the development of glutathione S-transferase placental form (GST-P)-positive foci in the liver. The results provided concrete evidence for the existence of no effect levels for hepatocarcinogenicity of MeIQx either in presence or absence of ethanol and, therefore, for a practical threshold for this genotoxic carcinogen. PMID- 16698720 TI - Absence of beta-catenin alteration in hepatic tumors induced by p-nitroanisole in Crj:BDF1 mice. AB - In the present study, beta-catenin localization in hepatocellular neoplasms and hepatoblastomas, induced by oral administration of p-Nitroanisole (pNA) in Crj:BDF1 for 2 years, was evaluated by immunohistochemistry along with genetic alterations in exon 2 of beta-catenin by the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) approach. Genomic DNA was isolated from paraffin sections of a total of 53 liver tumors. Immunohistochemical analysis revealed no abnormal accumulation of the beta-catenin protein in any of the cases. No mutations (0/13), 20% silent mutations (3/15) and 8% silent plus 12% functional mutations (2 + 3/25), not in the multiple phosphorylation sites of beta-catenin, were observed in hepatocellular adenomas, carcinomas and hepatoblastomas, respectively. The results indicate that beta-catenin does not play an important role in development of hepatic tumors induced by pNA in Crj:BDF1 mice. PMID- 16698721 TI - Retinal and peripheral nerve toxicity induced by the administration of a pan cyclin dependent kinase (cdk) inhibitor in mice. AB - Cyclin-dependent kinases (cdks) play a crucial role in cell cycle regulation and are considered promising targets for cancer therapy. Intravenous administration of AG-012986, a pan-cyclin-dependent kinase inhibitor (cdk(i)), resulted in unexpected retinal and peripheral nerve toxicity in mice. AG-012986 was administered daily to CD-1 or B6C3F1 mice for 5 consecutive days. Mice were euthanized 24 h after the last dose (study day 6) or after a 21-day post-dose period (study day 26). Compound related microscopic findings were seen in the sciatic nerves (axonal degeneration) of both strains and in the retina (retinal degeneration/atrophy) of CD-1 mice only after the post-dose period. Although retinal degeneration/atrophy was not detected by routine histology in mice euthanized on day 6, apoptotic retinal cells were evident at this time using TUNEL assay. To our knowledge retinal or peripheral nerve toxicity secondary to the administration of cdk(i)s has not been previously reported. Although the pathogenesis of these lesions is unclear, the toxicities may reflect the unique profile of cdk inhibition, off-target kinase inhibition or receptor binding, or metabolism/distribution properties of AG-012986. Multi-targeted-inhibitors may interfere with cdks and other kinases involved in a wide range of functions other than cell cycle regulation, which could result in unexpected toxicities that may hinder their clinical applications. PMID- 16698724 TI - The nose revisited: a brief review of the comparative structure, function, and toxicologic pathology of the nasal epithelium. AB - The nose is a very complex organ with multiple functions that include not only olfaction, but also the conditioning (e.g., humidifying, warming, and filtering) of inhaled air. The nose is also a "scrubbing tower" that removes inhaled chemicals that may be harmful to the more sensitive tissues in the lower tracheobronchial airways and pulmonary parenchyma. Because the nasal airway may also be a prime target for many inhaled toxicants, it is important to understand the comparative aspects of nasal structure and function among laboratory animals commonly used in inhalation toxicology studies, and how nasal tissues and cells in these mammalian species may respond to inhaled toxicants. The surface epithelium lining the nasal passages is often the first tissue in the nose to be directly injured by inhaled toxicants. Five morphologically and functionally distinct epithelia line the mammalian nasal passages--olfactory, respiratory, squamous, transitional, and lymphoepithelial--and each nasal epithelium may be injured by an inhaled toxicant. Toxicant-induced epithelial lesions in the nasal passages of laboratory animals (and humans) are often site-specific and dependent on the intranasal regional dose of the inhaled chemical and the sensitivity of the nasal epithelial tissue to the specific chemical. In this brief review, we present examples of nonneoplastic epithelial lesions (e.g., cell death, hyperplasia, metaplasia) caused by single or repeated exposure to various inhaled chemical toxicants. In addition, we provide examples of how nasal maps may be used to record the character, magnitude and distribution of toxicant-induced epithelial injury in the nasal airways of laboratory animals. Intranasal mapping of nasal histopathology (or molecular and biochemical alterations to the nasal mucosa) may be used along with innovative dosimetric models to determine dose/response relationships and to understand if site-specific lesions are driven primarily by airflow, by tissue sensitivity, or by another mechanism of toxicity. The present review provides a brief overview of comparative nasal structure, function and toxicologic pathology of the mammalian nasal epithelium and a brief discussion on how data from animal toxicology studies have been used to estimate the risk of inhaled chemicals to human health. PMID- 16698725 TI - Nasal dosimetry of inhaled gases and particles: where do inhaled agents go in the nose? AB - The anatomical structure of the nasal passages differs significantly among species, affecting airflow and the transport of inhaled gases and particles throughout the respiratory tract. Since direct measurement of local nasal dose is often difficult, 3-dimensional, anatomically accurate, computational models of the rat, monkey, and human nasal passages were developed to estimate regional transport and dosimetry of inhaled material. The computational models predicted that during resting breathing, a larger portion of inspired air passed through olfactory-lined regions in the rat than in the monkey or human. The models also predicted that maximum wall mass flux (mass per surface area per time) of inhaled formaldehyde in the nonsquamous epithelium was highest in monkeys (anterior middle turbinate) and similar in rats and humans (dorsal medial meatus in the rat and mid-septum in the human, near the squamous/nonsquamous epithelial boundary in both species). For particles that are 5 microm in aerodynamic diameter, preliminary simulations at minute volume flow rates predicted nasal deposition efficiencies of 92%, 11% and 25% in the rat, monkey, and human, respectively, with more vestibular deposition in the rat than in the monkey or human. Estimates such as these can be used to test hypotheses about mechanisms of toxicity and supply species-specific information for risk assessment, thus reducing uncertainty in extrapolating animal data to humans. PMID- 16698726 TI - Molecular biology of the nasal airways: how do we assess cellular and molecular responses in the nose? AB - We summarize studies herein that relate to use of molecular techniques to assess mechanism of toxicant and carcinogen action on the nasal mucosa. Specifically, we present the results of an in vivo mutagenesis assay with the herbicide alachlor, which causes olfactory mucosal tumors in rats following dietary administration. A positive response was found in olfactory mucosa after 3 mo of treatment. There was no increase in mutant frequency in the adjacent nasal respiratory mucosa or in liver, which are both non-target tissues for alachlor carcinogenesis. We also summarize previous findings of gene expression studies. One on these was a GeneChip experiment aimed at elucidating the mechanism of alachlor olfactory carcinogenesis, wherein we found that oxidative stress and gelatinase genes were upregulated early in the carcinogenic process, while genes consistent with activation of Wnt signaling were activated later in the carcinogenic process. The final example presented summarizes the results of a microarray experiment designed to identify novel olfactory genes involved in the plasticity of the olfactory mucosa. Those studies identified novel olfactory mucosal genes including Sgpl1 and Pon1. In each instance, precise sampling is emphasized and proper controls are discussed, and examples of independent means of validation of genomics experiments are presented. PMID- 16698727 TI - Types and patterns of response in the larynx following inhalation. AB - The laryngeal mucosa responds to insult similarly to other epithelial tissues but the response depends on location within the larynx since important anatomic differences exist, even within rodent species. Although dogs and nonhuman primates are also utilized for inhalation toxicology studies, little published information is available regarding sites of injury from inhaled toxicants in these species. Accurate and consistent fixation, trimming, and microtomy of laryngeal sections allow the pathologist to compare the morphology of laryngeal mucosa from exposed and control animals and detect and interpret subtle differences resulting from inhalation exposure. There are anatomic landmarks that are keys to providing consistent sections through important areas of the laryngeal mucosa. Repeated inhalation of toxic concentrations of chemicals, drugs, or environmental contaminants induces a wide range of responses, depending on the physical properties and concentration of the toxic substance and duration of exposure. Responses include edema, acute to chronic inflammation, fibrosis, mucosal ulceration, degeneration, and necrosis. Attempts at repair include regeneration, hyperplasia, squamous metaplasia, hyperkeratosis, and neoplasia. Awareness of normal histology and zones of transition from squamous to respiratory epithelium in different areas of the larynx in different species is critical to avoid confusing normal epithelium with metaplasia or hyperplasia. Microscopic examination of laryngeal mucosa from animals exposed via inhalation and necropsied following a recovery period provides the opportunity to determine the degree of regression or progression of exposure-induced laryngeal lesions. PMID- 16698728 TI - Design-based sampling and quantitation of the respiratory airways. AB - Design-based quantitation of the nasal cavity, larynx and tracheobronchial conducting airways after exposure to inhaled toxicants requires complete measurement of all respiratory airways or appropriate sampling followed by morphometric measurements. In vivo imaging (MRI or CT) of the nasal cavity, larynx and conducting airways provides anatomical detail of all the airways down to the distal airways. Since inhaled toxicants show predictable deposition patterns in the airways, identification and sampling of conducting airways becomes essential in a precise toxicological evaluation. Lengths, diameters and luminal surface areas can be directly measured on fixed specimens using a stereomicroscope. Estimates of cell numbers, extracellular matrix volumes and vessel/nerve lengths per airway or epithelial basal laminar surface are estimated stereologically. Selected airways are cut into smaller pieces using a "fractionator" for uniform sampling of the airways. Cell numbers are estimated using a "disector." Volumes are estimated using point probes, while length and surface areas are estimated by isotropically oriented sections with plane and line probes; an approach free of assumptions of shape, size or spatial orientation. True biological variance and the average sampling variance of the stereological measurement define the minimal sampling required to achieve precise estimates of the nasal cavity, larynx and conducting airways. PMID- 16698729 TI - A 'best practices' approach to neuropathologic assessment in developmental neurotoxicity testing--for today. AB - A key trait of developmental neurotoxicants is their ability to cause structural lesions in the immature nervous system. Thus, neuropathologic assessment is an essential element of developmental neurotoxicity (DNT) studies that are designed to evaluate chemically-induced risk to neural substrates in young humans. The guidelines for conventional DNT assays have been established by regulatory agencies to provide a flexible scaffold for conducting such studies; recent experience has launched new efforts to update these recommendations. The present document was produced by an ad hoc subcommittee of the Society of Toxicologic Pathology (STP) tasked with examining conventional methods used in DNT neuropathology in order to define the 'best practices' for dealing with the diverse requirements of both national (EPA) and international (OECD) regulatory bodies. Recommendations (including citations for relevant neurobiological and technical references) address all aspects of the DNT neuropathology examination: study design; tissue fixation, collection, processing, and staining; qualitative and quantitative evaluation; statistical analysis; proper control materials; study documentation; and personnel training. If followed, these proposals will allow pathologists to meet the need for a sound risk assessment (balanced to address both regulatory issues and scientific considerations) in this field today while providing direction for the research needed to further refine DNT neuropathology 'best practices' in the future. PMID- 16698732 TI - Prophylaxis for and treatment of deep venous thrombosis after stroke: the Post Stroke Rehabilitation Outcomes Project (PSROP). AB - Deep venous thrombosis (DVT) is a common and preventable complication after a stroke. Although the treatment of DVT is simple and straightforward, its prevention remains controversial. The Post-Stroke Rehabilitation Outcomes Project (PSROP) database was used to describe the incidence and temporal sequence of DVT and trends in the prevention and treatment of DVT. Of the 1,161 patients in the PSROP database, 383 (32.99%) patients without DVT and 8 (0.69%) with DVT had no documented orders for anticoagulant medications. Sixty-five (5.60%) patients had DVTs during the inpatient rehabilitation facility stay. Of 10 (0.86%) patients with DVTs in the common femoral vein, 4 (40%) were diagnosed within 24 hours of admission. Nine (90%) of these 10 patients were classified as moderate or severe strokes. All patients with common femoral DVT received appropriate therapy. Although much is known about the prevention, diagnosis, and treatment of poststroke DVT, clinicians need to learn and apply treatment protocols to prevent DVTs and allow more quality time for rehabilitation. PMID- 16698733 TI - Antiplatelet and anticoagulant medication usage during stroke rehabilitation: the Post-Stroke Rehabilitation Outcomes Project (PSROP). AB - Secondary prevention of strokes is an important issue during the admission to the inpatient rehabilitation facility (IRF). There are many clinical practice guidelines with strong levels of evidence that address the secondary prophylaxis of strokes. The Post-Stroke Rehabilitation Outcomes Project (PSROP) database was used to describe the frequency that antiplatelet and/or anticoagulant medications are prescribed for the secondary prophylaxis of a stroke. Of the 1,161 participants in the PSROP, 890 (76.66%) had nonhemorrhagic strokes. Of the participants with nonhemorrhagic strokes, 169 (18.99%) did not receive any antiplatelet or anticoagulant medication. Of 717 participants who did not have an embolic event, 140 (19.5%) did not receive a salicylate, antiplatelet agent, or warfarin. Of 173 participants who had an embolic event, 29 (16.8%) did not receive a salicylate, antiplatelet agent, or warfarin. Unless patients have any medical contraindications to these medications, they should receive these evidence-based treatments for secondary stroke prophylaxis. As more stroke survivors receive antiplatelet and/or anticoagulant medications, it is more likely that the incidence of recurrent strokes will decrease. PMID- 16698734 TI - Antihypertensive medication usage during stroke rehabilitation: the Post-Stroke Rehabilitation Outcomes Project (PSROP). AB - Blood pressure management is an important issue for the primary prevention, acute management, and secondary prevention of a stroke. Rehabilitation professionals need to consider the timing in lowering blood pressures in stroke survivors, the types of medications that should be used in managing hypertension, and the target pressures to achieve. The Post-Stroke Rehabilitation Outcomes Project (PSROP) database was used to describe the types of antihypertensive medications prescribed to stroke survivors, compare prescription patterns with current practice guidelines of the management of hypertension after a stroke, and determine whether systolic and diastolic blood pressures decrease during admissions to inpatient rehabilitation facilities (IRFs). Of the 1,161 patients in the PSROP database, the most commonly prescribed antihypertensive medications were the angiotensin-converting enzyme antagonists and angiotensin II antagonists, followed by beta-blockers, calcium-channel blockers and diuretics, adrenergics (alpha-blockers), and other (minoxidil, hydralazine) medications. Systolic, diastolic, and mean arterial pressures declined in participants during admissions to IRFs. However, blood pressures were significantly higher throughout IRF stays in participants receiving antihypertensive medications when compared to those not receiving antihypertensive medications. Rehabilitation professionals need to be cognizant of the relationship between stroke and hypertension, clinical practice guidelines that provide evidence-based management tools for hypertension, and patient issues that may hinder the effective treatment of hypertension. PMID- 16698736 TI - Usage of pain medications during stroke rehabilitation: the Post-Stroke Rehabilitation Outcomes Project (PSROP). AB - Pain remains one of the most common, yet most challenging, medical problems in health care today, and it is one of the most common complications that occurs after a stroke. Pain can affect the course of stroke rehabilitation adversely, and it occasionally may be a cause for transfer back to an acute care hospital. The Post-Stroke Rehabilitation Outcomes Project (PSROP) database was used to describe the incidence of pain by body location and trends in the use of different classifications of medications to treat pain. Of the 1,122 participants in the PSROP database, the most common locations of pain in stroke survivors were the head, leg, back, and shoulder. The most frequently prescribed classifications of pain medications were other analgesics (acetaminophen and tramadol), followed by narcotic analgesics, non-steroidal antiinflammatory drugs (NSAIDs), anticonvulsants, and tricyclic antidepressants. After acetaminophen, the most frequently prescribed medications in each classification, respectively, include hydrocodone APAP, cox-2 inhibitors, gabapentin, and amitriptyline. Other frequently prescribed pain medications included sumatriptan (migraine analgesic), cyclobenzaprine (muscle relaxant), and baclofen (antispasticity muscle relaxant). Medications should be chosen based upon the medical condition causing pain, the ability of the stroke survivor to comply with administration of the medication, and the cost of the medication. Appropriate and timely treatments of painful conditions result in maximum function and the ability to lead active lives and maintain an adequate quality of life. PMID- 16698735 TI - Neurostimulant medication usage during stroke rehabilitation: the Post-Stroke Rehabilitation Outcomes Project (PSROP). AB - Motor recovery after a stroke depends upon many upon different modalities. Intensive therapy using compensatory and facilitatory techniques is the primary method to improve movement and function in affected extremities. However, medications used to modulate neurotransmitters may be useful in augmenting therapy approaches. The Post-Stroke Rehabilitation Outcomes Project (PSROP) database was used to describe the frequency of prescribing neurostimulant medications; the types of neurostimulant medications used; and how the use of neurostimulant medications affected rehabilitation length of stay, motor recovery, cognitive recovery, and discharge destination. Of the 1,161 patients in the PSROP database, 929 (80.0%) patients did not receive any treatment with methylphenidate, modafinil, levodopa, amantadine, or bromocriptine. Patients who received neurostimulant medications did not have any more significant changes in length of stay, motor recovery, cognitive recovery, or discharge destination than patients who did not receive neurostimulant medications. Much research needs to be completed before clinicians know precisely whether and how rehabilitation therapies and medications interact to assist in functional recovery. PMID- 16698737 TI - Collaborative rehabilitation goal setting. PMID- 16698738 TI - Rehabilitation medicine summit: building research capacity. AB - The general objective of the "Rehabilitation Medicine Summit: Building Research Capacity" was to advance and promote research in medical rehabilitation by making recommendations to expand research capacity. The five elements of research capacity that guided the discussions were 1) researchers; 2) research culture, environment, and infrastructure; 3) funding; 4) partnerships; and 5) metrics. The 100 participants included representatives of professional organizations, consumer groups, academic departments, researchers, governmental funding agencies, and the private sector. The small group discussions and plenary sessions generated an array of problems, possible solutions, and recommended actions. A post-Summit, multi-organizational initiative is called to pursue the agendas outlined in this report. PMID- 16698739 TI - Damage control surgery--the intensivist's role. AB - "Damage control" surgery has evolved during the past 20 years from an accepted surgical technique in the traumatized, moribund patient to an expanded role in critically ill, nontraumatized patients. Physicians caring for these patients in extremis have begun to recognize a pattern of severe physiologic derangement that prompts an abbreviated laparotomy after hemorrhage and contamination control. Emphasis then shifts from the operating theater to the intensive care unit, where the patient's physiologic deficits are corrected. Once these derangements have been resolved, the patient is taken back to the operating room for definitive, reconstructive surgical care. The purpose of this article is to review the concept of "damage control" in reference to the patient whose pathophysiologic depletion prompts the need for it. Resuscitation in the intensive care unit will be summarized, pitfalls will be identified, and treatment plans will be delineated. Complications such as abdominal compartment syndrome and difficult abdominal wall closures will also be discussed. PMID- 16698740 TI - Diagnosis of ventilator-associated pneumonia: focus on nonbronchoscopic techniques (nonbronchoscopic bronchoalveolar lavage, including mini-BAL, blinded protected specimen brush, and blinded bronchial sampling) and endotracheal aspirates. AB - The ideal diagnostic approach for ventilator-associated pneumonia currently is based on invasive procedures to obtain respiratory tract cultures. Given the lack of consensus and relatively poor acceptance of full bronchoscopic bronchoalveolar lavage (BAL) and protected specimen brush (PSB), less invasive procedures have been developed. We review the nonbronchoscopic procedures (nonbronchoscopic bronchoalveolar lavage, including mini-BAL, blinded protected specimen, and blinded bronchial sampling) and endotracheal aspiration. We provide a critique of the methods used, the types of catheters inserted, and the sample collection methods. Most studies were flawed in that antibiotic use before initiation of the procedure was not controlled. The variability of both the methods and the criteria for the gold standard in the numerous investigations show that these procedures are neither standardized nor proven to be accurate and often did not improve management. Pending future studies, use of endotracheal aspirates without the use of quantitation seems to be a reasonable approach for clinicians who are not committed to an invasive procedure. PMID- 16698741 TI - Perioperative effects and safety of nesiritide following cardiac surgery in children. AB - Nesiritide (Natrecor, Scios Inc), human B-type natriuretic peptide, has hemodynamic effects that may be beneficial in pediatric patients after cardiac surgery. Experience with nesiritide and pediatrics is limited. The purpose of this study was to evaluate perioperative effects and safety of nesiritide in pediatric cardiothoracic surgery. Seventeen patients with congenital heart disease undergoing cardiac surgery were given a loading dose (1 microg/kg) while on cardiopulmonary bypass (constant flow) followed by continuous infusion for 24 hours (0.01 microg/kg/min x 6 hours, then 0.02 microg/kg/min x 18 hours). A 7% decrease in mean blood pressure was seen following nesiritide loading dose on cardiopulmonary bypass. No patient required intervention for hypotension while receiving nesiritide load or infusion. Nesiritide load during surgery and continuous infusion after cardiac surgery in pediatric patients resulted in no significant hemodynamic compromise. PMID- 16698742 TI - Use of recombinant activated factor VII (Novoseven) in trauma and surgery: analysis of outcomes reported to an international registry. AB - The objective was to evaluate the efficacy and safety of recombinant activated factor VII in patients with massive bleeding. Forty-five patients with severe massive hemorrhage requiring>or= 14 transfusion units of packed red blood cells received recombinant activated factor VII. Postdrug blood loss and transfusion requirements were assessed, and mortality was compared with predicted outcomes. Blood loss was markedly reduced in 40 of 43 (93.0%) patients, and transfusion requirements decreased after recombinant activated factor VII administration. Mortality rate in trauma patients who had massive hemorrhage was significantly reduced compared with predictions using scoring systems. This may be associated with the use of recombinant activated factor VII. This study failed to demonstrate an improvement in surgical patients. The absence of concurrent controls prevents definitive conclusions regarding actual safety or efficacy of recombinant activated factor VII. PMID- 16698743 TI - Mechanical complications of central venous catheters. AB - We analyzed 385 consecutive central venous catheter (CVC) attempts over a 6-month period. All critically ill patients 18 years of age or older requiring a CVC were included. The rate of mechanical complications not including failure to place was 14%. Complications included failure to place the CVC (n = 86), arterial puncture (n = 18), improper position (n = 14), pneumothorax (n = 5 in 258 subclavian and internal jugular attempts), hematoma (n = 3), hemothorax (n = 1), and asystolic cardiac arrest of unknown etiology (n = 1). Male patients had a significantly higher complication rate than female patients (37% vs 27%, P = .04). The subclavian approach had a higher complication rate than the internal jugular or the femoral approach (39% vs 33% vs. 24%, P = .02). The complication rate increased with the number of percutaneous punctures, with a rate of 54% when more than 2 punctures were required. PMID- 16698744 TI - Acute pulmonary edema due to rosiglitazone use in a patient with diabetes mellitus. AB - Rosiglitazone is a peroxisome proliferator active receptor. gamma agonist, which increases insulin sensitivity in adipose tissue, muscle, and liver. Rosiglitazone is a member of the thiazolidinedione group, and because of its significantly positive effect on glycemic control, it is especially preferred in type 2 diabetic patients with a high cardiovascular disease risk. This drug, because of its decreasing effect on insulin resistance, is used alone or combined with type 2 diabetic drugs. A 73-year-old female patient was admitted to the emergency department with dyspnea, pink frothing phlegm, cyanosis, and tiredness. She was lethargic, uncooperative, and had no orientation. In arterial blood gases, hypoxemia and hypercapnia were found. She was taken to the general intensive care unit, and oxygen was applied via mask. The patient had a history of 10 years of diabetes mellitus, hypertension, and atherosclerotic cardiac disease, and she was using rosiglitazone for the past 6 weeks. Her chest x-ray was taken, and acute pulmonary edema was diagnosed. In her last echocardiography, which was performed 1 year before, no signs indicating cardiac failure and pleural effusion could be found. Therefore, it was concluded that pulmonary edema occurred as a complication of rosiglitazone use. After stabilizing the patient's vital signs, blood glucose levels, and lactate levels, medical treatment of diabetes mellitus was rearranged, and she was discharged on the seventh day after her admittance. In a patient with diabetes mellitus who has been admitted to the intensive care unit because of acute pulmonary edema, for differential diagnosis, use of rosiglitazone should be kept in mind during the determination of treatment. Therefore, the authors aim to discuss the effect of rosiglitazone on creating acute pulmonary edema with a case report presentation. PMID- 16698745 TI - Central venous catheterization: better and worse. PMID- 16698746 TI - The Montreal classification of inflammatory bowel disease: controversies, consensus, and implications. AB - In recent years, investigators have readdressed the complex issues involved in the classification of inflammatory bowel diseases. In 2003, a Working Party of investigators with an interest in the issues involved in disease subclassification was formed with the aim of summarising recent developments in disease classification and establishing an integrated clinical, molecular, and serological classification of inflammatory bowel disease. The results of the Working Party were reported at the 2005 Montreal World Congress of Gastroenterology. Here we highlight the key issues that have emerged from discussions of the Montreal Working Party and the relevance to clinical practice and research activities. PMID- 16698747 TI - Designing Spiegelmers to antagonise ghrelin. PMID- 16698748 TI - The putative role of endogenous and exogenous opiates in inflammatory bowel disease. PMID- 16698749 TI - Probiotics for Crohn's disease: what have we learned? PMID- 16698750 TI - Prospective evaluation of fluorouracil chemotherapy based on the genetic makeup of colorectal cancer. PMID- 16698751 TI - Prognosis in Budd Chiari syndrome after re-establishing hepatic venous drainage. PMID- 16698752 TI - An unusual cause and treatment for ascites after liver transplantation. PMID- 16698753 TI - Intermittent fever in a patient with apparent fatty liver. PMID- 16698754 TI - An unsuspected cause of chronic colitis. PMID- 16698755 TI - Chronic pancreatitis and dyspnoea. PMID- 16698756 TI - Molecular subtyping of gastric MALT lymphomas: implications for prognosis and management. PMID- 16698757 TI - Endoscopic evaluation for cytomegalovirus enterocolitis after allogeneic haematopoietic stem cell transplantation. PMID- 16698758 TI - Second gastric cancers among patients with primary sporadic and familial cancers in Sweden. PMID- 16698759 TI - Hyperhomocysteinaemia and vascular disease in liver patients. PMID- 16698760 TI - Percutaneous gall bladder aspiration as an alternative to laparoscopic cholecystectomy in Child-Pugh C cirrhotic patients with acute cholecystitis. PMID- 16698761 TI - Wireless capsule endoscopic finding in Cronkhite-Canada syndrome. PMID- 16698762 TI - Immunonutrition enhances the expression and secretion of mouse intestinal defensins. PMID- 16698763 TI - A new silybin-vitamin E-phospholipid complex improves insulin resistance and liver damage in patients with non-alcoholic fatty liver disease: preliminary observations. PMID- 16698764 TI - Is the periodic repetition of a coagulation check necessary during anti-hepatitis C virus therapy? PMID- 16698765 TI - Seventeen years after successful small bowel transplantation: long term graft acceptance without immune tolerance. PMID- 16698766 TI - White cell scanning has a role in imaging small and large bowel. PMID- 16698767 TI - Is this really the current status of small bowel transplantation in the UK? PMID- 16698768 TI - A unification of models for meta-analysis of diagnostic accuracy studies. AB - Studies of diagnostic accuracy require more sophisticated methods for their meta analysis than studies of therapeutic interventions. A number of different, and apparently divergent, methods for meta-analysis of diagnostic studies have been proposed, including two alternative approaches that are statistically rigorous and allow for between-study variability: the hierarchical summary receiver operating characteristic (ROC) model (Rutter and Gatsonis, 2001) and bivariate random-effects meta-analysis (van Houwelingen and others, 1993), (van Houwelingen and others, 2002), (Reitsma and others, 2005). We show that these two models are very closely related, and define the circumstances in which they are identical. We discuss the different forms of summary model output suggested by the two approaches, including summary ROC curves, summary points, confidence regions, and prediction regions. PMID- 16698769 TI - Averaged gene expressions for regression. AB - Although averaging is a simple technique, it plays an important role in reducing variance. We use this essential property of averaging in regression of the DNA microarray data, which poses the challenge of having far more features than samples. In this paper, we introduce a two-step procedure that combines (1) hierarchical clustering and (2) Lasso. By averaging the genes within the clusters obtained from hierarchical clustering, we define supergenes and use them to fit regression models, thereby attaining concise interpretation and accuracy. Our methods are supported with theoretical justifications and demonstrated on simulated and real data sets. PMID- 16698770 TI - Observations of amino acid gain and loss during protein evolution are explained by statistical bias. AB - The authors of a recent manuscript in "Nature" claim to have discovered "universal trends" of amino acid gain and loss in protein evolution. Here, we show that this universal trend can be simply explained by a bias that is unavoidable with the 3-taxon trees used in the original analysis. We demonstrate that a rigorously reversible equilibrium model, when analyzed with the same methods as the "Nature" manuscript, yields identical (and in this case, clearly erroneous) conclusions. A main source of the bias is the division of the sequence data into "informative" and "noninformative" sites, which favors the observation of certain transitions. PMID- 16698771 TI - Does CO2 permeate through aquaporin-1? AB - Aquaporins facilitate water permeation across biological membranes. Additionally, glycerol and other small neutral solutes are permeated by related aquaglyceroporins. The role of aquaporins in gas permeation has been a long standing and controversially discussed issue. We present an extensive set of atomistic molecular dynamics simulations that address the question of CO(2) permeation through human aquaporin-1. Free energy profiles derived from the simulations display a barrier of approximately 23 kJ/mol in the aromatic/arginine constriction region of the water pore, whereas a barrier of approximately 4 kJ/mol was observed for a palmitoyloleoylphosphatidylethanolamine lipid bilayer membrane. The results indicate that significant aquaporin-1-mediated CO(2) permeation is to be expected only in membranes with a low intrinsic CO(2) permeability. PMID- 16698772 TI - Determination of the orientational distribution and orientation factor for transfer between membrane-bound fluorophores using a confocal microscope. AB - Orientational fluorophores have been a useful tool in physical chemistry, biochemistry, and more recently structural biology due to the polarized nature of the light they emit and that fact that energy can be transferred between them. We present a practical scheme in which measurements of the intensity of emitted fluorescence can be used to determine limits on the mean and distribution of orientation of the absorption transition moment of membrane-bound fluorophores. We demonstrate how information about the orientation of fluorophores can be used to calculate the orientation factor kappa(2) required for use in FRET spectroscopy. We illustrate the method using images of AlexaFluor probes bound to MscL mechanosensitive transmembrane channel proteins in spherical liposomes. PMID- 16698773 TI - Dynamics of the WPD loop of the Yersinia protein tyrosine phosphatase. AB - The bacterial protein tyrosine phosphatase YopH is an essential virulence determinant in Yersinia spp., causing gastrointestinal diseases and the plague. Like eukaryotic PTPases, YopH catalyzes the hydrolysis of the phosphate moiety of phosphotyrosine within a highly conserved binding pocket, which is also characterized by the closure of the so-called "WPD loop" upon ligand binding. In this study, we investigate the conformational changes and dynamics of the WPD loop by molecular dynamics simulations. Consistent with experimental observations, our simulations show that the WPD loop of YopH is intrinsically flexible and fluctuates between the open and closed conformation with a frequency of approximately 4 ns for the apo, native protein. The region of helix alpha4 spanning loop 384-392, which has been revealed experimentally as a second substrate-binding site in YopH, is found to be highly associated with the WPD loop, stabilizing it in the closed, active conformation, and providing a structural basis for the cooperation of the second-substrate binding site in substrate recognition. Loop L4 (residues 323-327) is shown to be involved in a parallel, correlated motion mode with the WPD loop that contributes the stabilization of a more extended open conformation. In addition, we have simulated the loop reopening in the ligand-bound protein complex by applying the locally enhanced sampling method. Finally, the dynamic behavior of the WPD loop for the C403S mutant differs from the wild-type YopH remarkably. These results shed light on the role of the WPD loop in PTPase-mediated catalysis, and are useful in structure-based design for novel, selective YopH inhibitors as antibacterial drugs. PMID- 16698774 TI - Model of a putative pore: the pentameric alpha-helical bundle of SARS coronavirus E protein in lipid bilayers. AB - The coronavirus responsible for the severe acute respiratory syndrome contains a small envelope protein, E, with putative involvement in host apoptosis and virus morphogenesis. To perform these functions, it has been suggested that protein E can form a membrane destabilizing transmembrane (TM) hairpin, or homooligomerize to form a TM pore. Indeed, in a recent study we reported that the alpha-helical putative transmembrane domain of E protein (ETM) forms several SDS-resistant TM interactions: a dimer, a trimer, and two pentameric forms. Further, these interactions were found to be evolutionarily conserved. Herein, we have studied multiple isotopically labeled ETM peptides reconstituted in model lipid bilayers, using the orientational parameters derived from infrared dichroic data. We show that the topology of ETM is consistent with a regular TM alpha-helix. Further, the orientational parameters obtained unequivocally correspond to a homopentameric model, by comparison with previous predictions. We have independently confirmed that the full polypeptide of E protein can also aggregate as pentamers after expression in Escherichia coli. This interaction must be stabilized, at least partially, at the TM domain. The model we report for this pentameric alpha-helical bundle may explain some of the permabilizing properties of protein E, and should be the basis of mutagenesis efforts in future functional studies. PMID- 16698775 TI - Secondary structure and Pd(II) coordination in S-layer proteins from Bacillus sphaericus studied by infrared and X-ray absorption spectroscopy. AB - The S-layer of Bacillus sphaericus strain JG-A12, isolated from a uranium-mining site, exhibits a high metal-binding capacity, indicating that it may provide a protective function by preventing the cellular uptake of heavy metals and radionuclides. This property has allowed the use of this and other S-layers as self-assembling organic templates for the synthesis of nanosized heavy metal cluster arrays. However, little is known about the molecular basis of the metal protein interactions and their impact on secondary structure. We have studied the secondary structure, protein stability, and Pd((II)) coordination in S-layers from the B. sphaericus strains JG-A12 and NCTC 9602 to elucidate the molecular basis of their biological function and of the metal nanocluster growth. Fourier transform infrared spectroscopy reveals similar secondary structures, containing approximately 35% beta-sheets and little helical structure. pH-induced infrared absorption changes of the side-chain carboxylates evidence a remarkably low pK < 3 in both strains and a structural stabilization when Pd((II)) is bound. The COO( )-stretching absorptions reveal a predominant Pd((II)) coordination by chelation/bridging by Asp and Glu residues. This agrees with XANES and EXAFS data revealing oxygens as coordinating atoms to Pd((II)). The additional participation of nitrogen is assigned to side chains rather than to the peptide backbone. The topology of nitrogen- and carboxyl-bearing side chains appears to mediate heavy metal binding to the large number of Asp and Glu in both S-layers at particularly low pH as an adaptation to the environment from which the strain JG-A12 has been isolated. These side chains are thus prime targets for the design of engineered S layer-based nanoclusters. PMID- 16698776 TI - A newly designed microspectrofluorometer for kinetic studies on protein crystals in combination with x-ray diffraction. AB - We present a new design for a fluorescence microspectrophotometer for use in kinetic crystallography in combination with x-ray diffraction experiments. The FLUMIX device (Fluorescence spectroscopy to monitor intermediates in x-ray crystallography) is built for 0 degrees fluorescence detection, which has several advantages in comparison to a conventional fluorometer with 90 degrees design. Due to the reduced spatial requirements and the need for only one objective, the system is highly versatile, easy to handle, and can be used for many different applications. In combination with a conventional stereomicroscope, fluorescence measurements or reaction initiation can be performed directly in a hanging drop crystallization setup. The FLUMIX device can be combined with most x-ray sources, normally without the need of a specialized mechanical support. As a biological model system, we have used H-Ras p21 with an artificially introduced photo-labile GTP precursor (caged GTP) and a covalently attached fluorophore (IANBD amide). Using the FLUMIX system, detailed information about the state of photolyzed crystals of the modified H-Ras p21 (p21(mod)) could be obtained. Measurements in combination with a synchrotron beamline showed significant fluorescence changes in p21(mod) crystals even within a few seconds of x-ray exposure at 100 K. PMID- 16698777 TI - Mechanical behavior of cellulose microfibrils in tension wood, in relation with maturation stress generation. AB - A change in cellulose lattice spacing can be detected during the release of wood maturation stress by synchrotron x-ray diffraction experiment. The lattice strain was found to be the same order of magnitude as the macroscopic strain. The fiber repeat distance, 1.033 nm evaluated for tension wood after the release of maturation stress was equal to the conventional wood values, whereas the value before stress release was larger, corresponding to a fiber repeat of 1.035 nm, nearly equal to that of cotton and ramie. Interestingly, the fiber repeat varied from 1.033 nm for wood to 1.040 nm for algal cellulose, with an increasing order of lateral size of cellulose microfibrils so far reported. These lines of experiments demonstrate that, before the stress release, the cellulose was in a state of tension, which is, to our knowledge, the first experimental evidence supporting the assumption that tension is induced in cellulose microfibrils. PMID- 16698778 TI - On imposing detailed balance in complex reaction mechanisms. PMID- 16698779 TI - Viewing dynamic assembly of molecular complexes by multi-wavelength single molecule fluorescence. AB - Complexes of macromolecules that transiently self-assemble, perform a particular function, and then dissociate are a recurring theme in biology. Such systems often have a large number of possible assembly/disassembly intermediates and complex, highly branched reaction pathways. Measuring the single-step kinetic parameters in these reactions would help to identify the functionally significant pathways. We have therefore constructed a novel single-molecule fluorescence microscope capable of efficiently detecting the colocalization of multiple components in a macromolecular complex when each component is labeled using a different color fluorescent dye. In this through-objective excitation, total internal reflection instrument, the dichroic mirror conventionally used to spectrally segregate the excitation and emission pathways was replaced with small broadband mirrors. This design spatially segregates the excitation and emission pathways and thereby permits efficient collection of the spectral range of emitted fluorescence when three or more dyes are used. In a test experiment with surface-immobilized single-stranded DNA molecules, we directly monitored the time course of a hybridization reaction with three different oligonucleotides, each labeled with a different color dye. The experiment reveals which of the possible reaction intermediates were traversed by each immobilized molecule, measures the hybridization rate constants for each oligonucleotide, and characterizes kinetic interdependences of the reaction steps. PMID- 16698780 TI - Minor groove deformability of DNA: a molecular dynamics free energy simulation study. AB - The conformational deformability of nucleic acids can influence their function and recognition by proteins. A class of DNA binding proteins including the TATA box binding protein binds to the DNA minor groove, resulting in an opening of the minor groove and DNA bending toward the major groove. Explicit solvent molecular dynamics simulations in combination with the umbrella sampling approach have been performed to investigate the molecular mechanism of DNA minor groove deformations and the indirect energetic contribution to protein binding. As a reaction coordinate, the distance between backbone segments on opposite strands was used. The resulting deformed structures showed close agreement with experimental DNA structures in complex with minor groove-binding proteins. The calculated free energy of minor groove deformation was approximately 4-6 kcal mol(-1) in the case of a central TATATA sequence. A smaller equilibrium minor groove width and more restricted minor groove mobility was found for the central AAATTT and also a significantly ( approximately 2 times) larger free energy change for opening the minor groove. The helical parameter analysis of trajectories indicates that an easier partial unstacking of a central TA versus AT basepair step is a likely reason for the larger groove flexibility of the central TATATA case. PMID- 16698781 TI - A mechanism for copper inhibition of infectious prion conversion. AB - We employ ab initio electronic structure calculations to obtain two structural models for copper bound in the strongest binding site of the noninfectious form of the prion protein. The models are compatible with available experimental constraints from electron spin resonance data. The bending of the peptide backbone attendant with the copper binding is not compatible with the requisite straight beta-strand backbone structure for the same sequence contained in two recently proposed models of the prion protein structure in its infectious form. We hypothesize that copper binding at this site is protective against conversion to the infectious form, discuss experimental data that appear to support and conflict with our hypothesis, and propose tests using recombinant prion protein, genetically modified cultured neurons, and transgenic mice. PMID- 16698782 TI - The conformation of calsequestrin determines its ability to regulate skeletal ryanodine receptors. AB - Ca2+ efflux from the sarcoplasmic reticulum decreases when store Ca2+ concentration falls, particularly in skinned fibers and isolated vesicles where luminal Ca2+ can be reduced to very low levels. However ryanodine receptor activity in many single channel studies is higher when the luminal free Ca2+ concentration is reduced. We investigated the hypothesis that prolonged exposure to low luminal Ca2+ causes conformational changes in calsequestrin and deregulation of ryanodine receptors, allowing channel activity to increase. Lowering of luminal Ca2+ from 1 mM to 100 microM for several minutes resulted in conformational changes with dissociation of 65-75% of calsequestrin from the junctional face membrane. The calsequestrin remaining associated no longer regulated channels. In the absence of this regulation, ryanodine receptors were more active when luminal Ca2+ was lowered from 1 mM to 100 microM. In contrast, when ryanodine receptors were calsequestrin regulated, lowering luminal Ca2+ either did not alter or decreased activity. Ryanodine receptors are regulated by calsequestrin under physiological conditions where calsequestrin is polymerized. Since depolymerization occurs slowly, calsequestrin can regulate the ryanodine receptor and prevent excess Ca2+ release when the store is transiently depleted, for example, during high frequency activity or early stages of muscle fatigue. PMID- 16698783 TI - Calcium-sensitive downregulation of the transduction chain in rod photoreceptors of the rat retina. AB - In vertebrate rod outer segments phototransduction is suggested to be modulated by intracellular Ca. We aimed at verifying this hypothesis by recording saturated photosignals in the rat retina after single and double flashes of light and determining the time t(c) to the beginning of the signal recovery. The time course of Ca(i) after a flash was calculated from a change of the spatial Ca(2+) concentration profile recorded in the space between the rods. After single flashes t(c) increased linearly with the logarithm of flash intensity, confirming the assumption that t(c) is determined by deactivation of a single species X* in the phototransduction cascade. The photoresponse was shortened up to 45% if the test flash was preceded by a conditioning preflash. The shortening depended on the reduction of Ca(i) induced by the preflash. The data suggest that the phototransduction gain determining the amount of activated X* is regulated by a Ca(i)-dependent mechanism in a short time period (<800 ms) after the test flash. Lowering of Ca(i) by a preflash reduced the gain up to 20% compared to its value in a dark-adapted rod. The relation between phototransduction gain and Ca(i) revealed a K(1/2) value close to the dark level of Ca(i). PMID- 16698784 TI - Dissociation of biological catch-bond by periodic perturbation. AB - The analysis of the P-selectin/PSGL-1 catch-slip bond that is periodically driven by a detaching force predicts that in the frequency range on the order of 1 s(-1) the bond lifetime undergoes significant changes with respect to both frequency and amplitude of the force. The result indicates how variations in the heart rate could have a substantial effect on leukocyte and lymphoid cell transport and adhesion to endothelial cells and platelets during inflammatory processes. PMID- 16698785 TI - Fourier transform infrared spectroscopy provides a fingerprint for the tetramer and for the aggregates of transthyretin. AB - Transthyretin (TTR) is an amyloidogenic protein whose aggregation is responsible for several familial amyloid diseases. Here, we use FTIR to describe the secondary structural changes that take place when wt TTR undergoes heat- or high pressure-induced denaturation, as well as fibril formation. Upon thermal denaturation, TTR loses part of its intramolecular beta-sheet structure followed by an increase in nonnative, probably antiparallel beta-sheet contacts (bands at 1,616 and 1,686 cm(-1)) and in the light scattering, suggesting its aggregation. Pressure-induced denaturation studies show that even at very elevated pressures (12 kbar), TTR loses only part of its beta-sheet structure, suggesting that pressure leads to a partially unfolded species. On comparing the FTIR spectrum of the TTR amyloid fibril produced at atmospheric pressure upon acidification (pH 4.4) with the one presented by the native tetramer, we find that the content of beta-sheets does not change much upon fibrillization; however, the alignment of beta-sheets is altered, resulting in the formation of distinct beta-sheet contacts (band at 1,625 cm(-1)). The random-coil content also decreases in going from tetramers to fibrils. This means that, although part of the tertiary- and secondary-structure content of the TTR monomers has to be lost before fibril formation, as previously suggested, there must be a subsequent reorganization of part of the random-coil structure into a well-organized structure compatible with the amyloid fibril, as well as a readjustment of the alignment of the beta sheets. Interestingly, the infrared spectrum of the protein recovered from a cycle of compression-decompression at pD 5, 37 degrees C, is quite similar to that of fibrils produced at atmospheric pressure (pH 4.4), which suggests that high hydrostatic pressure converts the tetramers of TTR into an amyloidogenic conformation. PMID- 16698786 TI - Tryptophan rotamers as evidenced by X-ray, fluorescence lifetimes, and molecular dynamics modeling. AB - We investigated the native-state dynamics of the Bacillus caldolyticus cold-shock protein mutant Bc-Csp L66E, using fluorescence and appropriate molecular dynamics methods. Two fluorescence lifetimes were found, the amplitudes of which agree very well with tryptophan rotamer populations, obtained from parallel tempering calculations. Rotamer lifetimes were predicted by transition-state theory from high-temperature simulations. Transition pathways were extracted from the transition rates between individual rotameric states. The molecular dynamics also reveal the loop fluctuations in the native state. PMID- 16698787 TI - Viscoelastic study of the mechanical unfolding of a protein by AFM. AB - We have applied a dynamic force modulation technique to the mechanical unfolding of a homopolymer of immunoglobulin (Ig) domains from titin, (C47S C63S I27)5, [(I27)5] to determine the viscoelastic response of single protein molecules as a function of extension. Both the stiffness and the friction of the homopolymer system show a sudden decrease when a protein domain unfolds. The decrease in measured friction suggests that the system is dominated by the internal friction of the (I27)5 molecule and not solvent friction. In the stiffness-extension spectrum we detected an abrupt feature before each unfolding event, the amplitude of which decreased with each consecutive unfolding event. We propose that these features are a clear indication of the formation of the known unfolding intermediate of I27, which has been observed previously in constant velocity unfolding experiments. This simple force modulation AFM technique promises to be a very useful addition to constant velocity experiments providing detailed viscoelastic characterization of single molecules under extension. PMID- 16698788 TI - Overview of the structure of all-AT oligonucleotides: organization in helices and packing interactions. AB - We present the crystalline organization of 33 all-AT deoxyoligonucleotide duplexes, studied by x-ray diffraction. Most of them have very similar structures, with Watson-Crick basepairs and a standard average twist close to 36 degrees. The molecules are organized as parallel columns of stacked duplexes in a helical arrangement. Such organization of duplexes is very regular and repetitive: all sequences show the same pattern. It is mainly determined by the stacking of the terminal basepairs, so that the twist in the virtual TA base step between neighbor duplexes is always negative, approximately -22 degrees. The distance between the axes of parallel columns is practically identical in all cases, approximately 26 A. Interestingly, it coincides with that found in DNA viruses and fibers in their hexagonal phase. It appears to be a characteristic distance for ordered parallel DNA molecules. This feature is due to the absence of short range intermolecular forces, which are usually due to the presence of CG basepairs at the end of the oligonucleotide sequence. The duplexes apparently interact only through their diffuse ionic atmospheres. The results obtained can thus be considered as intermediate between liquid crystals, fibers, and standard crystal structures. They provide new information on medium range DNA-DNA interactions. PMID- 16698789 TI - The rotor tip inside a bearing of a thermophilic F1-ATPase is dispensable for torque generation. AB - F(1)-ATPase is an ATP-driven rotary molecular motor in which the central gamma subunit rotates inside a stator cylinder made of alpha(3)beta(3) subunits. To elucidate the role of rotor-stator interactions in torque generation, we truncated the gamma-subunit at its carboxyl terminus, which forms an alpha helix that penetrates deeply into the stator cylinder. We used an alpha(3)beta(3)gamma subcomplex of F(1)-ATPase derived from thermophilic Bacillus PS3 and expressed it in Escherichia coli. We could obtain purified subcomplexes in which 14, 17, or 21 amino-acid residues were deleted. The rotary characteristics of the truncated mutants, monitored by attaching a duplex of 0.49-microm beads to the gamma subunit, did not differ greatly from those of the wild-type over the ATP concentrations of 20 nM-2 mM, the most conspicuous effect being approximately 50% reduction in torque and approximately 70% reduction in the rate of ATP binding upon deletion of 21 residues. The ATP hydrolysis activity estimated in bulk samples was more seriously affected. The 21-deletion mutant, in particular, was >10-fold less active, but this is likely due to instability of this subcomplex. For torque generation, though not for rapid catalysis, most of the rotor-stator contacts on the deeper half of the penetrating portion of the gamma-subunit are dispensable. PMID- 16698790 TI - Genetic deletion of Rac1 GTPase reveals its critical role in actin stress fiber formation and focal adhesion complex assembly. AB - Rac1 is an intracellular signal transducer regulating a variety of cell functions. Previous studies by overexpression of dominant-negative or constitutively active mutants of Rac1 in clonal cell lines have established that Rac1 plays a key role in actin lamellipodia induction, cell-matrix adhesion, and cell anoikis. In the present studies, we have examined the cellular behaviors of Rac1 gene-targeted primary mouse embryonic fibroblasts (MEFs) after Cre recombinase-mediated deletion of Rac1 gene. Rac1-null MEFs became contracted and elongated in morphology and were defective in lamellipodia formation, cell spreading, cell-fibronectin adhesion, and focal contact formation in response to platelet-derived growth factor or serum. Unexpectedly, deletion of Rac1 also abolished actin stress fibers in the cells without detectable alteration of endogenous RhoA activity. Although the expression and/or activation status of focal adhesion complex components such as Src, FAK, and vinculin were not affected by Rac1 deletion, the number and size of adhesion plaques were significantly reduced, and the molecular complex between Src, FAK, and vinculin was dissembled in Rac1-null cells. Overexpression of an active RhoA mutant or ROK failed to rescue the stress fiber and adhesion plaque defects of the Rac1-null cells. Although Rac1 deletion caused a significant reduction in phospho-PAK1, AKT, and -ERK under serum stimulation, reconstitution of active PAK1, but not AKT or MEK1, was able to rescue the actin cytoskeleton and adhesion phenotypes of the Rac1-deficient cells. Furthermore, Rac1 deletion led to a marked increase in spontaneous apoptosis that could be rescued by active PAK1, AKT, or MEK1 expression. Our results obtained from gene-targeted primary MEFs indicate that Rac1 is essential not only for lamellipodia induction but also for the RhoA regulated actin stress fiber and focal adhesion complex formation and that Rac1 is involved in cell survival regulation through anoikis-dependent as well as independent mechanisms. PMID- 16698791 TI - The crystal structure of human geranylgeranyl pyrophosphate synthase reveals a novel hexameric arrangement and inhibitory product binding. AB - Modification of GTPases with isoprenoid molecules derived from geranylgeranyl pyrophosphate or farnesyl pyrophosphate is an essential requisite for cellular signaling pathways. The synthesis of these isoprenoids proceeds in mammals through the mevalonate pathway, and the final steps in the synthesis are catalyzed by the related enzymes farnesyl pyrophosphate synthase and geranylgeranyl pyrophosphate synthase. Both enzymes play crucial roles in cell survival, and inhibition of farnesyl pyrophosphate synthase by nitrogen containing bisphosphonates is an established concept in the treatment of bone disorders such as osteoporosis or certain forms of cancer in bone. Here we report the crystal structure of human geranylgeranyl pyrophosphate synthase, the first mammalian ortholog to have its x-ray structure determined. It reveals that three dimers join together to form a propeller-bladed hexameric molecule with a mass of approximately 200 kDa. Structure-based sequence alignments predict this quaternary structure to be restricted to mammalian and insect orthologs, whereas fungal, bacterial, archaeal, and plant forms exhibit the dimeric organization also observed in farnesyl pyrophosphate synthase. Geranylgeranyl pyrophosphate derived from heterologous bacterial expression is tightly bound in a cavity distinct from the chain elongation site described for farnesyl pyrophosphate synthase. The structure most likely represents an inhibitory complex, which is further corroborated by steady-state kinetics, suggesting a possible feedback mechanism for regulating enzyme activity. Structural comparisons between members of this enzyme class give deeper insights into conserved features important for catalysis. PMID- 16698792 TI - Apolipoprotein A-I assumes a "looped belt" conformation on reconstituted high density lipoprotein. AB - Apolipoprotein A-I (apoA-I) plays a central role in the reverse cholesterol transport pathway; however, the structural basis for its antiatherogenic effects remains poorly understood. Here we employ EPR spectroscopy and fluorescence resonance energy transfer to elucidate the conformation and relative alignment of apoA-I monomers on discoidal (9.4 nm) reconstituted high density lipoprotein (rHDL). EPR spectroscopy provided evidence for an extended helical secondary structure. Position 139 since it was the only residue examined to display a dynamic motional character consistent with a flexible loop structure. The EPR spectra of nitroxide probes at positions 133 and 146 exhibit spin coupling, indicating that these positions are proximal to an apoA-I paired counterpart on the perimeter of rHDL. fluorescence resonance energy transfer studies employing engineered apoA-I variants possessing a single tryptophan (energy donor) and/or a single cysteine (whose thiol moiety was covalently labeled with an extrinsic energy acceptor) provided evidence that paired apoA-I molecules around the perimeter of rHDL align in an extended antiparallel conformation. Taken together with the observation that the EPR spectra of nitroxide probes positioned at intervening sequence positions (134-145) do not exhibit spin coupling, this has led us to propose a "looped belt" model, wherein residues 133-146 comprise a flexible loop segment that confers to apoA-I an intrinsic ability to adapt its structure to accommodate changing particle lipid content. Specifically, in the looped belt model, with the exception of amino acids 134-145, apoA-I aligns with its counterpart in a helix 5-helix 5 registry, centered at position 139. PMID- 16698793 TI - Role of neuregulin-1/ErbB2 signaling in endothelium-cardiomyocyte cross-talk. AB - Neuregulin-1 (NRG-1), a cardioactive growth factor released from endothelial cells, has been shown to be indispensable for the normal function of the adult heart by binding to ErbB4 receptors on cardiomyocytes. In the present study, we have investigated to what extent ErbB2, the favored co-factor of ErbB4 for heterodimerization, participates in the cardiac effects of endothelium-derived NRG-1. In addition, in view of our previously described anti-adrenergic effects of NRG-1, we have studied which neurohormonal stimuli affect endothelial NRG-1 expression and release and how this may fit into a broader frame of cardiovascular physiology. Immunohistochemical staining of rat heart and aorta showed that NRG-1 expression was restricted to the endocardial endothelium and the cardiac microvascular endothelium (CMVE); by contrast, NRG-1 expression was absent in larger coronary arteries and veins and in aortic endothelium. In rat CMVE in culture, NRG-1 mRNA and protein expression was down-regulated by angiotensin II and phenylephrine and up-regulated by endothelin-1 and mechanical strain. CMVE-derived NRG-1 was shown to phosphorylate cardiomyocyte ErbB2, an event prevented by a 24-h preincubation of myocytes with monoclonal ErbB2 antibodies. Pretreating cardiomyocytes with these inhibitory anti-ErbB2 antibodies significantly attenuated CMVE-induced cardiomyocyte hypertrophy and abolished the protective actions of CMVE against cardiomyocyte apoptosis. Accordingly, ErbB2 signaling participated in the paracrine survival and growth controlling effects of NRG-1 on cardiomyocytes in vitro, explaining the cardiotoxicity of ErbB2 antibodies in patients. Cardiac NRG-1 synthesis occurs in endothelial cells adjacent to cardiac myocytes and is sensitive to factors related to the regulation of blood pressure. PMID- 16698794 TI - Spi-1/PU.1 oncoprotein affects splicing decisions in a promoter binding-dependent manner. AB - The expression of the Spi-1/PU.1 transcription factor is tightly regulated as a function of the hematopoietic lineage. It is required for myeloid and B lymphoid differentiation. When overexpressed in mice, Spi-1 is associated with the emergence of transformed proerythroblasts unable to differentiate. In the course of a project undertaken to characterize the oncogenic function of Spi-1, we found that Spi-1 interacts with proteins of the spliceosome in Spi-1-transformed proerythroblasts and participates in alternative splice site selection. Because Spi-1 is a transcription factor, it could be hypothesized that these two functions are coordinated. Here, we have developed a system allowing the characterization of transcription and splicing from a single target. It is shown that Spi-1 is able to regulate alternative splicing of a pre-mRNA for a gene whose transcription it regulates. Using a combination of Spi-1 mutants and Spi-1 dependent promoters, we demonstrate that Spi-1 must bind and transactivate a given promoter to favor the use of the proximal 5' alternative site. This establishes that Spi-1 affects splicing decisions in a promoter binding-dependent manner. These results provide new insight into how Spi-1 may act in the blockage of differentiation by demonstrating that it can deregulate gene expression and also modify the nature of the products generated from target genes. PMID- 16698795 TI - Phosphatidylethanolamine and monoglucosyldiacylglycerol are interchangeable in supporting topogenesis and function of the polytopic membrane protein lactose permease. AB - To determine the specific role lipids play in membrane protein topogenesis in vivo, the orientation with respect to the membrane bilayer of Escherichia coli lactose permease (LacY) transmembrane (TM) domains and their flanking extramembrane domains was compared after assembly in native membranes and membranes with genetically modified lipid content using the substituted cysteine accessibility method for determining TM domain mapping. LacY assembled in the absence of the major membrane lipid phosphatidylethanolamine (PE) does not carry out uphill transport of substrate and displays an inverted orientation for the N terminal six-TM domain helical bundle (Bogdanov, M., Heacock, P. N., and Dowhan, W. (2002) EMBO J. 21, 2107-2116). Strikingly, the replacement of PE in vivo by the foreign lipid monoglucosyldiacylglycerol (MGlcDAG), synthesized by the Acholeplasma laidlawii MGlcDAG synthase, restored uphill transport and supported the wild type TM topology of the N-terminal helical bundle of LacY. An interchangeable role in defining membrane protein TM domain orientation and supporting function is played by the two most abundant lipids, PE and MGlcDAG, in gram-negative and gram-positive bacteria, respectively. Therefore, these structurally diverse lipids endow the membrane with similar properties necessary for the proper organization of protein domains in LacY that are highly sensitive to lipids as topological determinants. PMID- 16698796 TI - Involvement of aquaporin 9 in osteoclast differentiation. AB - Aquaporins (water channels) selectively enhance water permeability of membranes. Since osteoclast differentiation includes a dramatic increase in cell volume, we hypothesize that aquaporin(s) is/are critical for the formation of the multinucleated osteoclast from its mononuclear precursor. Our studies employ two cell models, bone marrow macrophages (BMMs) and the murine macrophage-like cell line, RAW264.7, as osteoclast precursors. Receptor activator of nuclear factor kappaB (NF-kappaB) ligand (RANKL) and macrophage-colony-stimulating factor or RANKL alone were used to induce osteoclast differentiation in BMMs or RAW264.7 cells, respectively. We first used qualitative reverse transcription (RT)-PCR to examine which of the aquaporins are expressed in osteoclasts and in their precursor cells. Out of the 10 aquaporins examined, only aquaporin 9 (AQP9) was expressed in osteoclast-lineage cells. AQP9 has unique aqueous pore properties mediating the passage of a wide variety of non-charged solutes in addition to water. Western analyses using specific antibodies revealed a higher AQP9 level in RANKL-treated than in untreated cells. Quantitative real-time RT-PCR analyses also demonstrated higher AQP9 mRNA levels in RANKL-treated cells. Finally, we examined the effect of phloretin, an AQP9 inhibitor, on RANKL-induced osteoclast differentiation. Cells were incubated with RANKL for 5 days, and phloretin was added for the last 2 days, when most fusion occurs. A dramatic reduction in osteoclast size and in the number of nuclei per osteoclast was observed in cultures containing phloretin. The inhibitor did not have a significant effect on the number and size of mononuclear phagocytes in cultures not treated with RANKL. Our results suggest a role for AQP9 in osteoclast differentiation, specifically in the fusion process. PMID- 16698797 TI - Physical and functional association of human protein O-mannosyltransferases 1 and 2. AB - A defect of protein O-mannosylation causes congenital muscular dystrophy with brain malformation and structural eye abnormalities, so-called Walker-Warburg syndrome. Protein O-mannosylation is catalyzed by protein O-mannosyltransferase 1 (POMT1) and its homologue, POMT2. Coexpression of POMT1 and POMT2 is required to show O-mannosylation activity. Here we have shown that POMT1 forms a complex with POMT2 and the complex possesses protein O-mannosyltransferase activity. Results indicate that POMT1 and POMT2 associate physically and functionally in vivo. Recently, three mutations were reported in the POMT1 gene of patients who showed milder phenotypes than typical Walker-Warburg syndrome. We coexpressed these mutant POMT1s with POMT2 and found that none of them had any activity. However, all POMT1 mutants, including previously identified POMT1 mutants, coprecipitated with POMT2. These results indicate that the mutant POMT1s could form heterocomplexes with POMT2 but that such complexes are insufficient for enzymatic activity. PMID- 16698798 TI - Affinity of TatCd for TatAd elucidates its receptor function in the Bacillus subtilis twin arginine translocation (Tat) translocase system. AB - Twin arginine translocation (Tat) systems catalyze the transport of folded proteins across the bacterial cytosolic membrane or the chloroplast thylakoid membrane. In the Tat systems of Escherichia coli and many other species TatA-, TatB-, and TatC-like proteins have been identified as essential translocase components. In contrast, the Bacillus subtilis phosphodiesterase PhoD-specific system consists only of a pair of TatA(d)/TatC(d) proteins and involves a TatA(d) protein engaged in a cytosolic and a membrane-embedded localization. Because soluble TatA(d) was able to bind the twin arginine signal peptide of prePhoD prior to membrane integration it could serve to recruit its substrate to the membrane via the interaction with TatC(d). By analyzing the distribution of TatA(d) and studying the mutual affinity with TatC(d) we have shown here that TatC(d) assists the membrane localization of TatA(d). Besides detergent solubilized TatC(d), membrane-integrated TatC(d) showed affinity for soluble TatA(d). By using a peptide library-specific binding of TatA(d) to cytosolic loops of membrane protein TatC(d) was demonstrated. Depletion of TatC(d) in B. subtilis resulted in a drastic reduction of TatA(d), indicating a stabilizing effect of TatC(d) for TatA(d). In addition, the presence of the substrate prePhoD was the prerequisite for appropriate localization in the cytosolic membrane of B. subtilis as demonstrated by freeze-fracture experiments. PMID- 16698799 TI - Cell cycle-controlled interaction of nucleolin with the retinoblastoma protein and cancerous cell transformation. AB - Retinoblastoma protein (Rb) is a multifunctional tumor suppressor, frequently inactivated in certain types of human cancer. Nucleolin is an abundant multifunctional phosphoprotein of proliferating and cancerous cells, recently identified as cell cycle-regulated transcription activator, controlling expression of human papillomavirus type 18 (HPV18) oncogenes in cervical cancer. Here we find that nucleolin is associated with Rb in intact cells in the G1 phase of the cell cycle, and the complex formation is mediated by the growth-inhibitory domain of Rb. Association with Rb inhibits the DNA binding function of nucleolin and in consequence the interaction of nucleolin with the HPV18 enhancer, resulting in Rb-mediated repression of the HPV18 oncogenes. The intracellular distribution of nucleolin in epithelial cells is Rb-dependent, and an altered nucleolin localization in human cancerous tissues results from a loss of Rb. Our findings suggest that deregulated nucleolin activity due to a loss of Rb contributes to tumor development in malignant diseases, thus providing further insights into the molecular network for the Rb-mediated tumor suppression. PMID- 16698800 TI - N-terminally truncated WT1 protein with oncogenic properties overexpressed in leukemia. AB - WT1 was originally identified as an inactivated gene in Wilms tumor, a childhood kidney cancer. Alternative splicing of the WT1 transcript generates four major protein isoforms, each having different functional properties. Here we characterized a short transcript originating from a second promoter located within intron 1 of WT1. This 2.3-kb sWT1 transcript encodes a protein of approximately 35-37 kDa that retains intact DNA-binding and transactivation domains but lacks the 147 amino acids at the N terminus required for transcriptional repression. We found sWT1 to be a more potent transcriptional activator than WT1 for cyclin E and insulin-like growth factor 1 receptor promoters, which are normally repressed by WT1. The expression patterns of the sWT1 and WT1 transcripts differed slightly in various organs; we found sWT1 protein in tissue samples from adult testis and fetal kidney, with low-level expression in adult kidney as well. The sWT1 transcript, but not the full-length transcript, was over-expressed in the leukemia samples tested. sWT1-specific small interfering RNA retarded the proliferation of leukemia cell line K562 in vitro. Finally, sWT1 cooperated with Ras in transforming primary fibroblasts in vitro. Further studies are needed to clarify the oncogenic behavior of this isoform and to determine the mechanism underlying its up-regulation in leukemia and other forms of cancer. PMID- 16698801 TI - Functional characterization of Src-interacting Na/K-ATPase using RNA interference assay. AB - We have shown that the Na/K-ATPase and Src form a signaling receptor complex. Here we determined how alterations in the amount and properties of the Na/K ATPase affect basal Src activity and ouabain-induced signal transduction. Several alpha1 subunit knockdown cell lines were generated by transfecting LLC-PK1 cells with a vector expressing alpha1-specific small interference RNA. Although the alpha1 knockdown resulted in significant decreases in Na/K-ATPase activity, it increased the basal Src activity and tyrosine phosphorylation of focal adhesion kinase, a Src effector. Concomitantly it also abolished ouabain-induced activation of Src and ERK1/2. When the knockdown cells were rescued by a rat alpha1, both Na/K-ATPase activity and the basal Src activity were restored. In addition, ouabain was able to stimulate Src and ERK1/2 in the rescued cells at a much higher concentration, consistent with the established differences in ouabain sensitivity between pig and rat alpha1. Finally both fluorescence resonance energy transfer analysis and co-immunoprecipitation assay indicated that the pumping-null rat alpha1 (D371E) mutant could also bind Src. Expression of this mutant restored the basal Src activity and focal adhesion kinase tyrosine phosphorylation. Taken together, the new findings suggest that LLC-PK1 cells contain a pool of Src-interacting Na/K-ATPase that not only regulates Src activity but also serves as a receptor for ouabain to activate protein kinases. PMID- 16698802 TI - Escaping from the TGFbeta anti-proliferative control. AB - Transforming growth factor-beta (TGFbeta) has a crucial role in tissue homeostasis and disruption of the TGFbeta pathway has been implicated in many human diseases including cancer. As a potent inhibitor of epithelial cell proliferation, TGFbeta is a tumor suppressor. Tumor cells evade the antitumoral effect of TGFbeta, either by acquiring somatic mutations that blunt TGFbeta signaling or by selectively preventing the cytostatic responses to TGFbeta. During tumor progression, TGFbeta not only loses the anti-proliferative response but can also become an oncogenic factor. Recent work has provided insights into the specific molecular mechanisms involved in the loss of the TGFbeta anti proliferative response. This review is an overview of the mechanisms that lead to the impairment of the tumor-suppressive function of TGFbeta in cancer. The understanding of how the TGFbeta signal is disrupted in cancer might facilitate the design and development of rational and successful therapeutic strategies. PMID- 16698803 TI - Predictive factors of acute urinary retention requiring catheterization following 125I prostate brachytherapy. AB - OBJECTIVE: To analyze predictive factors of acute urinary retention requiring catheterization after (125)I prostate brachytherapy. METHODS: A group of 227 consecutive patients with localized prostate cancer were treated with (125)I prostate brachytherapy between September 2003 and December 2004. The clinical, treatment-related and dosimetric factors were evaluated for the need for catheterization owing to urinary retention. RESULTS: Twelve patients (5.3%) required catheterization. The median time to onset was 2 days after implantation (range 1-7 days). Univariate analysis demonstrated that pre-implant ultrasound prostate volume, number of seeds, number of needles and neoadjuvant hormonal manipulation were predictive for catheterization. In multivariate analysis, the number of needles and neoadjuvant hormonal manipulation were significant independent predictive factors for catheterization (P = 0.002 and 0.025, respectively). The risk of catheterization in the cluster in which the number of needles was >24 was 4.07 times as high as that in the cluster in which the number of needles was < or =24 [11.3% versus 3.0%, P = 0.020; 95% confidence interval (CI) 1.24-13.36], and the risk in the hormonally manipulated patients was 7.05 times as high as that in the hormone-naive patients (7.7% versus 1.2%, P = 0.034; 95% CI 0.89-55.64). CONCLUSION: Our data suggest that the number of needles and hormonal manipulation might be the strongest predictors for catheterization. PMID- 16698804 TI - Smoking, obesity, and their co-occurrence in the United States: cross sectional analysis. AB - OBJECTIVES: To describe the prevalence of obesity, smoking, and both health risk factors together among adults in the United States. DESIGN: Cross sectional analysis of a national health interview survey. SETTING: United States. PARTICIPANTS: 29 305 adults (aged > or = 18) in 2002. MAIN OUTCOME MEASURES: Prevalence of adults who are obese (body mass index > or = 30), who smoke, and who are obese and smoke. Prevalence was stratified by age, sex, ethnic group, education, and income. RESULTS: 23.5% of adults were obese, 22.7% smoked, and 4.7% smoked and were obese. CONCLUSIONS: Although the proportion of adults who smoke and are obese is relatively low, this subgroup is concentrated among lower socioeconomic groups. PMID- 16698805 TI - Injustice at work and incidence of psychiatric morbidity: the Whitehall II study. AB - BACKGROUND: Previous studies of organisational justice and mental health have mostly examined women and have not examined the effect of change in justice. AIM: To examine effects of change in the treatment of employees by supervisors (the relational component of organisational justice) on minor psychiatric morbidity, using a cohort with a large proportion of men. METHODS: Data are from the Whitehall II study, a prospective cohort of 10 308 white-collar British civil servants (3143 women and 6895 men, aged 35-55 at baseline) (Phase 1, 1985-88). Employment grade, relational justice, job demands, job control, social support at work, effort-reward imbalance, physical illness, and psychiatric morbidity were measured at baseline. Relational justice was assessed again at Phase 2 (1989-90). The outcome was cases of psychiatric morbidity by Phases 2 and 3 (1991-93) among participants case-free at baseline. RESULTS: In analyses adjusted for age, grade, and baseline physical illness, women and men exposed to low relational justice at Phase 1 were at higher risk of psychiatric morbidity by Phases 2 and 3. Adjustment for other psychosocial work characteristics, particularly social support and effort-reward imbalance, partially attenuated these associations. A favourable change in justice between Phase 1 and Phase 2 reduced the immediate risk (Phase 2) of psychiatric morbidity, while an adverse change increased the immediate and longer term risk (Phase 3). CONCLUSION: This study shows that unfair treatment by supervisors increases risk of poor mental health. It appears that the employers' duty to ensure that employees are treated fairly at work also has benefits for health. PMID- 16698806 TI - Work-family conflict as a risk factor for sickness absence. AB - OBJECTIVES: (1) To study both cross-sectional and prospective relationships between work-family conflict and sickness absence from work; (2) to explore the direction of the relationships between the different types of conflict (work-home interference and home-work interference) and sickness absence; and (3) to explore gender differences in the above relationships. METHODS: Data from the Maastricht Cohort Study were used with six months of follow up (5072 men and 1015 women at T6). Work-family conflict was measured with the Survey Work-Home Interference Nijmegen (SWING). Sickness absence was assessed objectively through individual record linkage with the company registers on sickness absence. RESULTS: In the cross-sectional analyses, high levels of work-family conflict, work-home interference, and home-work interference were all associated with a higher odds of being absent at the time of completing the questionnaire, after controlling for age and long term disease. Differences in average number of absent days between cases and non-cases of work-home interference were significant for men and most pronounced in women, where the average number of absent days over six months follow up was almost four days higher in women with high versus low-medium work-home interference. CONCLUSIONS: A clear relation between work-family conflict and sickness absence was shown. Additionally, the direction of work family conflict was associated with a different sickness absence pattern. Sickness absence should be added to the list of adverse outcomes for employees struggling to combine their work and family life. PMID- 16698807 TI - Prediction of sickness absence: development of a screening instrument. AB - OBJECTIVES: To develop a concise screening instrument for early identification of employees at risk for sickness absence due to psychosocial health complaints. METHODS: Data from the Maastricht Cohort Study on "Fatigue at Work" were used to identify items to be associated with an increased risk of sickness absence. The analytical procedures univariate logistic regression, backward stepwise linear regression, and multiple logistic regression were successively applied. For both men and women, sum scores were calculated, and sensitivity and specificity rates of different cut-off points on the screening instrument were defined. RESULTS: In women, results suggested that feeling depressed, having a burnout, being tired, being less interested in work, experiencing obligatory change in working days, and living alone, were strong predictors of sickness absence due to psychosocial health complaints. In men, statistically significant predictors were having a history of sickness absence, compulsive thinking, being mentally fatigued, finding it hard to relax, lack of supervisor support, and having no hobbies. A potential cut-off point of 10 on the screening instrument resulted in a sensitivity score of 41.7% for women and 38.9% for men, and a specificity score of 91.3% for women and 90.6% for men. CONCLUSIONS: This study shows that it is possible to identify predictive factors for sickness absence and to develop an instrument for early identification of employees at risk for sickness absence. The results of this study increase the possibility for both employers and policymakers to implement interventions directed at the prevention of sickness absence. PMID- 16698808 TI - Update of mortality and cancer incidence in the Australian petroleum industry cohort. AB - OBJECTIVES: To update the analysis of the cohort mortality and cancer incidence study of employees in the Australian petroleum industry. METHODS: Employees of Australian Institute of Petroleum member companies were enrolled in the cohort in four industry-wide surveys between 1981 and 1999. Mortality of 16,547 males and 1356 females was determined up to 31 December 2001 and cancer incidence to 31 December 2000. Cause specific mortality and cancer incidence were compared with those of the Australian population by means of standardised mortality ratios (SMRs) and standardised incidence ratios (SIRs). Associations between increased incidence of specific cancers and employment in the petroleum industry were tested by trends according to period of first employment, duration of employment, latency, and hydrocarbon exposure, adjusting for personal smoking history where appropriate. RESULTS: There was a significant elevation of the incidence of mesothelioma (SIR 1.77, 95% CI 1.05 to 2.79), melanoma (SIR 1.37, 95% CI 1.19 to 1.58), and prostate cancer (SIR 1.18, 95% CI 1.04 to 1.34). The SIRs of all leukaemias and of acute non-lymphocytic leukaemia (ANLL) were not significantly different from unity, but all 11 ANLL cases were clustered in the middle to high hydrocarbon exposure categories. Tanker drivers had a significantly elevated incidence of kidney cancer (12 cases v 5.84 expected, SIR 2.05, 95% CI 1.06 to 3.59). Lung cancer incidence was significantly reduced (SIR 0.69, 95% CI 0.57 to 0.83) CONCLUSIONS: Most cases of mesothelioma are probably related to past exposure to asbestos in refineries. No occupational cause has been identified for the excess of melanoma, or prostatic or bladder cancer. The possibility of a causal relationship between cancer of the kidney and hydrocarbon exposure warrants further study. It is uncertain whether benzene exposures, particularly past levels of exposure, have been high enough to cause ANLL. PMID- 16698810 TI - Chronic fatigue syndrome-like caseness as a predictor of work status in fatigued employees on sick leave: four year follow up study. AB - OBJECTIVE: To assess whether CFS-like caseness (meeting the criteria for chronic fatigue syndrome (CFS)) predicts work status in the long term. METHODS: Prospective study in a sample of fatigued employees absent from work. Data were collected at baseline and four years later, and included CFS-like caseness and work status (inactive work status and full work incapacity). RESULTS: CFS-like cases at baseline were three times more likely to be unable to work at follow up than fatigued employees who did not meet CFS criteria at baseline (ORs 3-3.3). These associations grew even stronger when demographic and clinical confounders were controlled for (ORs 3.4-4.4). CONCLUSION: A CFS-like status (compared to non CFS fatigue) proved to be a strong predictor of an inactive work status and full work incapacity in the long term. Since little is known about effective interventions that prevent absenteeism and work incapacity or facilitate return to work in subjects with chronic fatigue, there is a great need for powerful early interventions that restore or preserve the ability to work, especially for workers who meet criteria for CFS. PMID- 16698809 TI - Association of ventricular arrhythmias detected by implantable cardioverter defibrillator and ambient air pollutants in the St Louis, Missouri metropolitan area. AB - BACKGROUND: It has previously been reported that the risk of ventricular arrhythmias is positively associated with ambient air pollution among patients with implantable cardioverter defibrillators (ICD) in Boston. AIMS: To assess the association of community exposures to air pollution with ventricular arrhythmias in a cohort of ICD patients in metropolitan St Louis, Missouri. METHODS: ICD detected episodes reported during clinical follow up were abstracted and reviewed by an electrophysiologist to identify ventricular arrhythmias. A total of 139 ventricular arrhythmias were identified among 56 patients. A case-crossover design was used with control periods matched on weekday and hour of the day within the same calendar month. Conditional logistic regression models were adjusted for temperature, barometric pressure, and relative humidity in the 24 hours preceding the event. RESULTS: There was a significant (24%, 95% CI 7% to 44%) increase in risk of ventricular arrhythmias associated with each 5 ppb increase in mean sulphur dioxide and non-significantly increased risk (22%, 95% CI -6% to 60%; and 18%, 95% CI -7% to 50%) associated with increases in nitrogen dioxide (6 ppb) and elemental carbon (0.5 microg/m3), respectively in the 24 hours before the arrhythmia. CONCLUSIONS: These results provide evidence of an association between ventricular arrhythmias and ambient air pollutants in St Louis. This is consistent with previous results from Boston, although the pollutants responsible for the increased risk are different. PMID- 16698811 TI - Fertility and exposure to solvents among families in the Agricultural Health Study. AB - BACKGROUND: Several studies have reported associations between solvent exposure and reduced female fertility, but the evidence is inconclusive for male fertility. OBJECTIVES: To investigate the impact of solvent exposure on subfertility among families of male licensed pesticide applicators in the Agricultural Health Study cohort. METHODS: The couples enrolled between 1993 and 1997. Cross-sectional questionnaire information on work tasks was used to assess exposure to solvents. The data were limited to couples (wife aged less than 40 years) with an attempt at pregnancy in the last four years (n = 2112). RESULTS: Twenty eight per cent of the couples were defined as subfertile (not conceiving a pregnancy after at least 12 months of unprotected intercourse, regardless of whether or not a pregnancy ultimately occurred). Adjusted subfertility odds ratios (OR) for exposure to solvents were calculated with logistic regression. Female (OR 1.42, 95% CI 1.15 to 1.75) and male exposure to solvents (OR 1.21 (95% CI 0.93 to 1.57) for monthly exposure and 1.40 (95% CI 0.97 to 2.03) for daily or weekly exposure) were associated with subfertility. In farming, spouses may share or exchange jobs. To account for potential dual exposure, variables for parental exposure (either parent exposed or both parents exposed) were also defined. Both were strongly associated with subfertility (OR 1.62 (95% CI 1.20 to 2.17) and OR 2.10 (95% CI 1.22 to 3.60), respectively). CONCLUSIONS: Solvents may impair fertility of either gender, though the evidence for female effects is stronger than for male effects. PMID- 16698812 TI - The role of ascorbic acid in the control of flowering time and the onset of senescence. AB - Ascorbic acid (AA) is not only an important antioxidant, it also appears to link flowering time, developmental senescence, programmed cell death, and responses to pathogens through a complex signal transduction network. The biological activity of AA is defined by its oxidation and subsequent regeneration into the reduced form. Some studies suggest that the total endogenous level of AA influences induction of flowering and senescence. Both processes require the co-ordinated regulation of gene expression, which is mediated by various phytohormones. For example, gibberellins and salicylic acid are known to promote flowering, but inhibit or retard senescence in Arabidopsis. Ethylene and abscisic acid accelerate senescence. Ascorbic acid serves as an important co-factor for the synthesis of some of these hormones. Therefore, it is assumed that AA affects phytohormone-mediated signalling processes during the transition from the vegetative to the reproductive phase and the final stage of development, senescence. This review summarizes recent reports that investigate the effect of AA on flowering time and the onset of senescence. An attempt was made to bring these findings in context with previously characterized flowering and senescence pathways and a model is proposed that may explain how AA influences flowering and senescence both under long- and short-day conditions in Arabidopsis. PMID- 16698813 TI - Effect of ascorbate and its oxidation products on H2O2 production in cell suspension cultures of Picea abies and in the absence of cells. AB - Dehydroascorbate and traces of ascorbate were present apoplastically in living spruce (Picea abies) twigs. Since the proposed apoplastic ascorbate degradation pathway contains several steps that possibly generate H(2)O(2), the effects of ascorbate and some of its degradation products were tested on apoplastic H(2)O(2) concentrations in a cell culture of P. abies as a model and on non-enzymic H(2)O(2) production in vitro. Ascorbate scavenged H(2)O(2) in the culture medium of lignin-producing Picea cells and in spent and boiled spent medium; in the presence of Cu(2+) or fresh medium, ascorbate led to the non-enzymic generation of H(2)O(2). Preparations of dehydroascorbate (the initial oxidation-product of ascorbate), and diketogulonate (the hydrolysis-product of dehydroascorbate) induced H(2)O(2) accumulation both non-enzymically and enzymically in Picea cell suspensions. Paper electrophoresis showed that the dehydroascorbate and diketogulonate preparations contained several degradation products; some of these probably contributed to H(2)O(2) production and/or scavenging in these experiments, and would also do so in vivo. These results indicate a complex ability of apoplastic ascorbate, dehydroascorbate, diketogulonate, and further products to modulate H(2)O(2) concentrations, with potential consequences for the control of growth, development and lignification. PMID- 16698814 TI - Controlled levels of salicylic acid are required for optimal photosynthesis and redox homeostasis. AB - Sudden exposure of plants to high light (HL) leads to metabolic and physiological disruption of the photosynthetic cells. Changes in ROS content, adjustment of photosynthetic processes and the antioxidant pools and, ultimately, gene induction are essential components for a successful acclimation to the new light conditions. The influence of salicylic acid (SA) on plant growth, short-term acclimation to HL, and on the redox homeostasis of Arabidopsis thaliana leaves was assessed here. The dwarf phenotype displayed by mutants with high SA content (cpr1-1, cpr5-1, cpr6-1, and dnd1-1) was less pronounced when these plants were grown in HL, suggesting that the inhibitory effect of SA on growth was partly overcome at higher light intensities. Moreover, higher SA content affected energy conversion processes in low light, but did not impair short-term acclimation to HL. On the other hand, mutants with low foliar SA content (NahG and sid2-2) were impaired in acclimation to transient exposure to HL and thus predisposed to oxidative stress. Low and high SA levels were strictly correlated to a lower and higher foliar H(2)O(2) content, respectively. Furthermore high SA was also associated with higher GSH contents, suggesting a tight correlation between SA, H(2)O(2) and GSH contents in plants. These observations implied an essential role of SA in the acclimation processes and in regulating the redox homeostasis of the cell. Implications for the role of SA in pathogen defence signalling are also discussed. PMID- 16698815 TI - Antioxidative enzymes from chloroplasts, mitochondria, and peroxisomes during leaf senescence of nodulated pea plants. AB - In this work the influence of the nodulation of pea (Pisum sativum L.) plants on the oxidative metabolism of different leaf organelles from young and senescent plants was studied. Chloroplasts, mitochondria, and peroxisomes were purified from leaves of nitrate-fed and Rhizobium leguminosarum-nodulated pea plants at two developmental stages (young and senescent plants). In these cell organelles, the activity of the ascorbate-glutathione cycle enzymes ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), and glutathione reductase (GR), and the ascorbate and glutathione contents were determined. In addition, the total superoxide dismutase (SOD) activity, the pattern of mitochondrial and peroxisomal NADPH-generating dehydrogenases, some of the peroxisomal photorespiratory enzymes, the glyoxylate cycle and oxidative metabolism enzymes were also analysed in these organelles. Results obtained on the metabolism of cell organelles indicate that nodulation with Rhizobium accelerates senescence in pea leaves. A considerable decrease of the ascorbate content of chloroplasts, mitochondria, and peroxisomes was found, and in these conditions a metabolic conversion of leaf peroxisomes into glyoxysomes, characteristic of leaf senescence, took place. PMID- 16698816 TI - Photoinhibition of manganese enzymes: insights into the mechanism of photosystem II photoinhibition. AB - Evidence has recently been presented that photoinhibition of photosystem II (PSII) is triggered by absorption of light by the oxygen-evolving manganese cluster. To get insight into the effects of light on enzymes containing manganese or other transition metal cofactors, the photosensitivities of Mn catalase, Mn superoxide dismutase, the haem (Fe)-containing bovine liver catalase, and CuZn superoxide dismutase were investigated. Glucose oxidase was studied as an example of an enzyme that does not have a metal cofactor. Sensitivities of these five enzymes to UVC, UVA, and visible light were compared in anaerobic conditions. The Mn(III)-oxo-Mn(III)-containing Mn catalase was found to be more sensitive to both visible and UV light than bovine liver catalase. Furthermore, the action spectrum of photoinhibition of Mn catalase was found to be fairly similar to that of photoinhibition of PSII. The Mn(II)-containing Mn superoxide dismutase was sensitive to UVC light and somewhat sensitive to UVA light, while only UVC light caused some inhibition of CuZn superoxide dismutase. Glucose oxidase was the least photosensitive of the enzymes studied. The photosensitivity of Mn enzymes supports the hypothesis that the oxygen-evolving manganese complex of PSII can be damaged by UV and visible light absorbed by its Mn(III) or Mn(IV) ions. PMID- 16698817 TI - Reactive oxygen and nitrogen species and glutathione: key players in the legume Rhizobium symbiosis. AB - Several reactive oxygen and nitrogen species (ROS/RNS) are continuously produced in plants as by-products of aerobic metabolism or in response to stresses. Depending on the nature of the ROS and RNS, some of them are highly toxic and rapidly detoxified by various cellular enzymatic and non-enzymatic mechanisms. Whereas plants have many mechanisms with which to combat increased ROS/RNS levels produced during stress conditions, under other circumstances plants appear to generate ROS/RNS as signalling molecules to control various processes encompassing the whole lifespan of the plant such as normal growth and development stages. This review aims to summarize recent studies highlighting the involvement of ROS/RNS, as well as the low molecular weight thiols, glutathione and homoglutathione, during the symbiosis between rhizobia and leguminous plants. This compatible interaction initiated by a molecular dialogue between the plant and bacterial partners, leads to the formation of a novel root organ capable of fixing atmospheric nitrogen under nitrogen-limiting conditions. On the one hand, ROS/RNS detection during the symbiotic process highlights the similarity of the early response to infection by pathogenic and symbiotic bacteria, addressing the question as to which mechanism rhizobia use to counteract the plant defence response. Moreover, there is increasing evidence that ROS are needed to establish the symbiosis fully. On the other hand, GSH synthesis appears to be essential for proper development of the root nodules during the symbiotic interaction. Elucidating the mechanisms that control ROS/RNS signalling during symbiosis could therefore contribute in defining a powerful strategy to enhance the efficiency of the symbiotic interaction. PMID- 16698818 TI - The role of stigma peroxidases in flowering plants: insights from further characterization of a stigma-specific peroxidase (SSP) from Senecio squalidus (Asteraceae). AB - Angiosperm stigmas have long been known to exhibit high levels of peroxidase activity when they are mature and most receptive to pollen but the biological function of stigma peroxidases is not known. A novel stigma-specific class III peroxidase gene, SSP (stigma-specific peroxidase) expressed exclusively in the stigmas of Senecio squalidus L. (Asteraceae) has recently been identified. Expression of SSP is confined to the specialized secretory cells (papillae) that compose the stigma epidermis. The literature on stigma peroxidases and hypotheses on their function(s) is reviewed here before further characterization of SSP and an attempt to determine its function are described. It is shown that SSP is localized to cytoplasmic regions of stigmatic papillae and also to the surface of these cells, possibly as a component of the pellicle, a thin layer of condensed protein typical of "dry" stigmas. Enzyme assays on recombinant SSP showed it to be a peroxidase with a preference for diphenolic substrates (ABTS and TMB) and a pH optimum of approximately 4.5. In such assays the peroxidase activity of SSP was low when compared with horseradish peroxidase. To explore the function of SSP and other stigmatic peroxidases, levels of reactive oxygen species (ROS) in stigmas of S. squalidus were investigated. Relatively large amounts of ROS, principally H(2)O(2), were detected in S. squalidus stigmas where most ROS/H(2)O(2) was localized to the stigmatic papillae, the location of SSP. These observations are discussed in the context of possible functions for SSP, other peroxidases, and ROS in the stigmas of angiosperms. PMID- 16698819 TI - Rapid hydropassive opening and subsequent active stomatal closure follow heat induced electrical signals in Mimosa pudica. AB - In Mimosa pudica L., heat stimulation triggers leaflet folding in local, neighbouring and distant leaves. Stomatal movements were observed microscopically during this folding reaction and electrical potentials, chlorophyll fluorescence, and leaf CO(2)/H(2)O-gas exchange were measured simultaneously. Upon heat stimulation of a neighbouring pinna, epidermal cells depolarized and the stomata began a rapid and pronounced transient opening response, leading to an approximately 2-fold increase of stomatal aperture within 60 s. At the same time, net CO(2) exchange showed a pronounced transient decrease, which was followed by a similar drop in photochemical quantum yield at photosystem (PS) II. Subsequently, CO(2)-gas exchange and photochemical quantum yield recovered and stomata closed partly or completely. The transient and fast stomatal opening response is interpreted as a hydropassive stomatal movement caused by a sudden loss of epidermal turgor. Thus, epidermal cells appear to respond in a similar manner to heat-induced signals as the pulvinar extensor cells. The subsequent closing of the stomata confirms earlier reports that stomatal movements can be induced by electrical signals. The substantial delay (several minutes) of guard cell turgor loss compared with the immediate response of the extensor and epidermal cells suggests a different, less direct mechanism for transmission of the propagating signal to the guard cells. PMID- 16698820 TI - Physiological and biochemical responses of fruit exocarp of tomato (Lycopersicon esculentum Mill.) mutants to natural photo-oxidative conditions. AB - Photo-oxidative stress was imposed under natural solar radiation on exposed and shaded sections of detached fruit of immature green tomato (Lycopersicon esculentum Miller = Solanum lycopersicum L.) mutants (anthocyanin absent, beta carotene, Delta, and high pigment-1) and their nearly isogenic parents ('Ailsa Craig' and 'Rutgers'). After 5 h exposure to high solar irradiance, either with or without ultraviolet (UV) radiation, surface colour changes, pigment composition, photosynthetic efficiency, antioxidant metabolites and enzyme activities, and selected flavonoids and antioxidant proteins in exocarp tissue were evaluated. The imposed photo-oxidative stress reproduced the symptoms observed on attached fruit. Both high temperature and solar irradiance caused fruit surface discoloration with faster degradation of chlorophyll (Chl) than carotenoids (Car), leading to an increase in the Car/Chl ratio. Surface bleaching was mostly caused by visible light, whereas elevated temperatures were mostly responsible for the inactivation of photosynthesis, measured as decreased F(v)/F(m). Ascorbate, glutathione, and total soluble protein concentrations decreased in the exocarp as the duration of exposure increased. Specific activities of superoxide dismutase, ascorbate peroxidase, dehydroascorbate reductase, monodehydroascorbate reductase (MDHAR), glutathione reductase (GR), and catalase increased with exposure, suggesting that these proteins were conserved during the imposed stress. GR protein expression remained stable during the imposed stress, whereas, MDHAR protein expression increased. Quercetin and kaempferol concentrations increased rapidly upon exposure, but not to UV radiation, suggesting rapid photo-protection in response to visible light; however, naringenin synthesis was not induced. The apparent increased tolerance of hp-1 fruit is discussed. PMID- 16698821 TI - National variations in the provision of cardiac services in the United Kingdom: second report of the British Cardiac Society Working Group, 2005. PMID- 16698822 TI - Maternal diabetes and the fetal heart. AB - Maternal diabetes mellitus significantly affects the fetal heart and fetal placental circulation in both structure and function. The influence of pre conceptional diabetes begins during embryonic development in the first trimester, with altered cardiac morphogenesis and placental development. It continues to have an influence on the fetal circulation through the second and third trimesters and into the perinatal and neonatal period. PMID- 16698824 TI - Images in cardiology. Traumatic left ventricular true aneurysm: echocardiographic, MRI, and intraoperative images. PMID- 16698823 TI - Arrhythmogenic right ventricular dysplasia, a cell adhesion cardiomyopathy: insights into disease pathogenesis from preliminary genotype--phenotype assessment. AB - Arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVC) is a genetically determined heart muscle disorder presenting clinically with even lethal ventricular arrhythmias, particularly in the young and athletes. It is reported familial with recessive and most commonly dominant inheritance. Disease-causing genes are increasingly recognised among desmosomal proteins plakoglobin, desmoplakin, plakophilin2, and desmoglein2 displaying phenotypic heterogeneity. Mutations in the plakoglobin and desmoplakin genes have been identified to underlie recessive ARVC associated with woolly hair and palmoplantar keratoderma (Naxos disease), while mutations in plakophilin2, desmoglein2 as well as desmoplakin have been identified to underlie the dominant non-syndromic form. Preliminary genotype-phenotype assessment indicates that mutations affecting the outer dense plaque of desmosome (desmoglein2, plakoglobin, plakophilin2 and the N terminal of desmoplakin) result in ARVC with the ordinary described phenotype. However, mutations at the inner dense plaque, particularly affecting the desmin binding site of desmoplakin, may result in ARVC with predominantly left ventricular involvement and clinical overlapping with dilated cardiomyopathy. The interesting finding of abnormal distribution of plakoglobin, independently of the primarily affected protein, might suggest a common pathway for plakoglobin in ARVC pathogenesis. PMID- 16698825 TI - Sir Brian Barratt-Boyes KBE. PMID- 16698826 TI - Emerging medical treatments for aortic stenosis: statins, angiotensin converting enzyme inhibitors, or both? AB - Aortic stenosis is the most common adult heart valve condition seen in the Western world and its incidence continues to rise. No established disease modifying treatments retard progression of the stenotic process. Recent insights into the pathogenesis of calcific aortic stenosis suggest that the disease mimics atherosclerosis. The natural history and progression of calcific aortic stenosis are described with particular emphasis on new and emerging medical treatments that may modify the disease process. In particular, statins and angiotensin converting enzyme inhibitors appear to hold promise but definitive evidence from large clinical trials is awaited. PMID- 16698827 TI - Images in cardiology. A common cause of secondary hypertension: coarctation of the aorta. PMID- 16698829 TI - Images in cardiology. Angiographic appearance of "tumour blush" produced by a large right atrial myxoma. PMID- 16698830 TI - Images in cardiology. Unusual acute coronary syndrome. PMID- 16698831 TI - Images in cardiology. Brugada syndrome unmasked by a shift of right precordial leads. PMID- 16698832 TI - Images in cardiology. An unusual mass related to the right atrium. PMID- 16698833 TI - Images in cardiology. Obstructed left coronary cusp as a cause of left main coronary artery obstruction and severe aortic regurgitation. PMID- 16698834 TI - Images in cardiology. Twiddler's syndrome in a patient with an implantable cardioverter defibrillator. PMID- 16698835 TI - Images in cardiology. Infective pseudoaneurysm of a ruptured sinus of Valsalva as an unusual cause of myocardial infarction by compression of the right coronary artery. PMID- 16698836 TI - Images in cardiology. Thrombotic obliteration of the right ventricular cavity. PMID- 16698837 TI - Plasma concentrations of D-dimer predict mortality in acute type A aortic dissection. PMID- 16698838 TI - Images in cardiology. Stent structure after kissing balloon inflation visualised by 64 slice computed tomography. PMID- 16698839 TI - Increased erythropoietin production after myocardial infarction in mice. PMID- 16698840 TI - Images in cardiology. Atrial myocarditis: a possible cause of idiopathic enlargement of bilateral atria. PMID- 16698841 TI - Role of B-type natriuretic peptide (BNP) and NT-proBNP in clinical routine. PMID- 16698842 TI - Basics of cardiac pacing: selection and mode choice. PMID- 16698843 TI - The genetics of heart attack. PMID- 16698844 TI - Stamps in cardiology. Society of perfusionists of Great Britain and Ireland. PMID- 16698845 TI - Hospital consumption of antibiotics in 15 European countries: results of the ESAC Retrospective Data Collection (1997-2002). AB - OBJECTIVES: To collect reliable, comparable and publicly available data on hospital use of antibiotics in Europe aggregated at the national level (1997 2002). METHODS: Consumption data of systemic antibiotics in Anatomical Therapeutic Chemical (ATC) class J01 were collected and expressed in defined daily doses (DDD) per 1000 inhabitants per day. Valid data for 2002 were available for 15 countries, and 6 year trends for 10 countries. Comparison with ambulatory care (AC) consumption data was possible in 14 countries. RESULTS: In 2002, median national hospital antibiotic consumption in Europe was 2.1 DDD/1000 inhabitants/day in Europe, ranging from 3.9 in Finland and France to 1.3 in Norway and Sweden. Hospital care (HC) consumption as a proportion of total antibiotic consumption ranged from 17.8% to 6.4%. The consumption of hospital specific antibiotics ranged from 0.43 DDD/1000 inhabitants/day in Greece and 0.08 in Sweden. Six-year trends in consumption were stable, except for rising co amoxiclav exposure and more rapid market penetration of new antibiotics (e.g. levofloxacin) in some countries. There was a strong, positive correlation between the extent of antibiotic use in AC and in HC (Spearman coefficient 0.745; P = 0.002), both for overall use and for use of five main classes (not macrolides and 'others'). In contrast to AC consumption no substantial seasonal variation in consumption was observed. CONCLUSIONS: It was cumbersome but feasible to collect ecological data on hospital antibiotic consumption in a set of 15 European countries on a retrospective basis, illustrating substantial cross-national variations in the extent and distribution of exposure to antibiotics in hospital care. PMID- 16698846 TI - Activity of mecillinam against AmpC beta-lactamase-producing Escherichia coli. PMID- 16698847 TI - Antimicrobial susceptibility of Bifidobacterium strains from humans, animals and probiotic products. AB - OBJECTIVES: The aim of this study was to assess the antimicrobial susceptibility of a taxonomically diverse set of Bifidobacterium strains to different classes of antimicrobial agents using a recently described medium. METHODS: The susceptibility of 100 strains encompassing 11 bifidobacterial species originating from humans, animals and probiotic products to 12 antimicrobial agents was tested by agar overlay disc diffusion. Based on these results, one or two strains per species were selected for susceptibility testing to nine antibiotics by broth microdilution using the Lactic acid bacteria Susceptibility test Medium (LSM) supplemented with cysteine. The genotypic basis of atypical tetracycline resistance was further characterized using PCR, Southern blotting and partial sequencing. RESULTS: Based on the distribution of inhibition zone diameters and MIC values, all strains tested were susceptible to amoxicillin, chloramphenicol, erythromycin, quinupristin/dalfopristin, rifampicin and vancomycin. Our data also reinforce earlier observations indicating that bifidobacteria are intrinsically resistant to gentamicin, sulfamethoxazole and polymyxin B. Susceptibility to trimethoprim, trimethoprim/sulfamethoxazole, ciprofloxacin, clindamycin, tetracycline and minocycline was variable. The tet(W) gene was responsible for tetracycline resistance in 15 strains including 7 probiotic isolates belonging to the taxa Bifidobacterium animalis subsp. lactis and Bifidobacterium bifidum. This gene was present in a single copy on the chromosome and did not appear to be associated with the conjugative transposon TnB1230 previously found in tet(W) containing Butyrivibrio fibrisolvens. CONCLUSIONS: The use of the LSM + cysteine medium allowed us to discriminate between intrinsic and atypical resistance properties of bifidobacteria and sets the scene for future definition of epidemiological cut-off values for all important Bifidobacterium species. The presence of an acquired tet(W) gene in several probiotic product isolates stresses the need for a minimal safety evaluation during the selection of Bifidobacterium strains for probiotic use. PMID- 16698848 TI - Comment on: empirical antibiotic monotherapy for febrile neutropenia: systematic review and meta-analysis of randomized controlled trials. PMID- 16698849 TI - Urinary bromide and breathing zone concentrations of 1-bromopropane from workers exposed to flexible foam spray adhesives. AB - 1-Bromopropane (1-BP) has been marketed as an alternative for ozone depleting solvents and suspect carcinogens and is in aerosol products, adhesives and solvents used for metal, precision and electronics cleaning. Toxicity of 1-BP is poorly understood, but it may be a neurologic, reproductive and hematologic toxin. Sparse exposure information prompted this exposure assessment study using air sampling, and measurement of urinary metabolites. Mercapturic acid conjugates are excreted in urine from 1-BP metabolism involving removal of bromide (Br) from the propyl group. One research objective was to evaluate the utility of urinary Br analysis for assessing 1-BP exposure using a relatively inexpensive, commercially available method. Complete 48 h urine specimens were obtained from 30 workers on two consecutive days at two facilities using 1-BP adhesives to construct polyurethane foam seat cushions and from seven unexposed control subjects. All of the workers' urine was collected into composite samples representing three daily time intervals (at work; after work but before bedtime; and upon wake-up) and analyzed for Br ion by inductively coupled plasma-mass spectrometry. Full-shift breathing zone samples were collected for 1-BP on Anasorb carbon molecular sieve sorbent tubes and analyzed by gas chromatography flame ionization detection via NIOSH method 1025. Geometric mean (GM) breathing zone concentrations of 1-BP were 92 parts per million (p.p.m.) for adhesive sprayers and 11 p.p.m. for other jobs. For sprayers, urinary Br concentrations ranged from 77 to 542 milligrams per gram of creatinine [mg (g-cr)(-1)] at work; from 58 to 308 mg (g-cr)(-1) after work; and from 46 to 672 mg (g-cr)(-1) in wake up samples. Pre-week urinary Br concentrations for sprayers were substantially higher than for the non-sprayers and controls, with GMs of 102, 31 and 3.8 mg (g cr)(-1), respectively. An association of 48 h urinary Br concentration with 1-BP exposure was statistically significant (r(2) = 0.89) for all jobs combined. This study demonstrates that urinary elimination is an important excretion pathway for 1-BP metabolism, and Br may be a useful biomarker of exposure. PMID- 16698850 TI - Mediators and modulators of pulmonary arterial hypertension. AB - Pulmonary hypertension (PH), defined as a mean pulmonary arterial (PA) pressure of >25 mmHg at rest or >30 mmHg during exercise, is characterized by a progressive and sustained increase in pulmonary vascular resistance that eventually leads to right ventricular failure. Clinically, PH may result from a variety of underlying diseases (Table 1 and Refs. 50, 113, 124). Pulmonary arterial hypertension (PAH) may be familial (FPAH) or sporadic (idiopathic, IPAH), formerly known as primary pulmonary hypertension, i.e., for which there is no demonstrable cause. More often, PAH is due to a variety of identifiable diseases including scleroderma and other collagen disorders, liver disease, human immunodeficiency virus, and the intake of appetite-suppressant drugs such as phentermine and fenfluramine (72). Other, more common, causes of PAH include left ventricular failure (perhaps the most common cause), valvular lesions, chronic pulmonary diseases, sleep-disordered breathing, and prolonged residence at high altitude. This classification, now widely accepted, was first proposed at a meeting in Evian, France, in 1998, and modified in Venice, Italy, in 2003 (124). PMID- 16698851 TI - Choosing the frequency of deep inflation in mice: balancing recruitment against ventilator-induced lung injury. AB - Low tidal volume (Vt) ventilation is protective against ventilator-induced lung injury but can promote development of atelectasis. Periodic deep inflation (DI) can open the lung, but if delivered too frequently may cause damage via repeated overdistention. We therefore examined the effects of varying DI frequency on lung mechanics, gas exchange, and biomarkers of injury in mice. C57BL/6 males were mechanically ventilated with positive end-expiratory pressure (PEEP) of 2 cmH2O for 2 h. One high Vt group received a DI with each breath (HV). Low Vt groups received 2 DIs after each hour of ventilation (LV) or 2 DIs every minute (LVDI). Control groups included a nonventilated surgical sham and a group receiving high Vt with zero PEEP (HVZP). Respiratory impedance was measured every 4 min, from which tissue elastance (H) and damping (G) were derived. G and H rose progressively during LV and HVZP, but returned to baseline after hourly DI during LV. During LVDI and HV, G and H remained low and gas exchange was superior to that of LV. Bronchoalveolar lavage fluid protein was elevated in HV and HVZP but was not different between LV and LVDI. Lung tissue IL-6 and IL-1beta levels were elevated in HVZP and lower in LVDI compared with LV. We conclude that frequent DI can safely improve gas exchange and lung mechanics and may confer protection from biotrauma. Differences between LVDI and HV suggest that an optimal frequency range of DI exists, within which the benefits of maintaining an open lung outweigh injury incurred from overdistention. PMID- 16698852 TI - Ectopic expression of C/EBPalpha in the lung epithelium disrupts late lung development. AB - The lung develops from the endoderm through a process of branching morphogenesis. This process is highly active during the pseudoglandular stage of lung development and continues into the canalicular stage, resulting in the formation of terminal sacs. CCAAT/enhancer binding proteins (C/EBPs) are transcription factors regulating central aspects of differentiation and proliferation. We report here the developmental expression of C/EBPalpha, -beta, and -delta in the lung. C/EBPalpha exhibits a dynamic expression pattern and is first detected during the late pseudoglandular stage. At this stage, expression is observed in a subset of epithelial cells in the distal parts of the branching tubules. The expression of C/EBPalpha is confined to nonproliferating cells. To examine the role of C/EBPalpha in lung development, we generated transgenic mice ectopically expressing C/EBPalpha in the lung epithelium using the human surfactant protein C promoter. Lungs from these mice were of normal size but exhibited a phenotype characterized by fewer and larger developing epithelial tubules, indicating that the branching process was affected. No effects on overall proliferation or cellular differentiation were observed. When this phenotype was compared with that of mice carrying a targeted mutation of the Cebpa gene, the Cebpa-/- mice exhibited a similar developmental phenotype. In conclusion, our results show a role for C/EBPalpha in lung development and suggest a function in the later stages of lung branching morphogenesis. PMID- 16698853 TI - Simvastatin causes endothelial cell apoptosis and attenuates severe pulmonary hypertension. AB - Severe pulmonary hypertension (SPH) is characterized by precapillary arteriolar lumen obliteration, dramatic right ventricular hypertrophy, and pericardial effusion. Our recently published rat model of SPH recapitulates major components of the human disease. We used this model to develop new treatment strategies for SPH. SPH in rats was induced using VEGF receptor blockade in combination with chronic hypoxia. A large variety of drugs used in this study, including anticancer drugs (cyclophosphamide and paclitaxel), the angiotensin-converting enzyme inhibitor lisinopril, the antiangiogenic agent thalidomide, and the peroxisome proliferator-actived receptor-gamma agonist PGJ2, failed to decrease mean pulmonary artery pressure (PAP) or right ventricular hypertrophy. In contrast, treatment of rats with established SPH with simvastatin markedly reduced mean PAP and right ventricular hypertrophy, and this reduction was associated with caspase-3 activation and pulmonary microvascular endothelial cell apoptosis. Simvastatin partially restored caveolin-1, caveolin-2, and phospho caveolin expression in vessel walls. In rat primary pulmonary microvascular endothelial cells, simvastatin induced caspase 3 activation and Rac 1 expression while suppressing Rho A and attenuated levels of Akt and ERK phosphorylation. We conclude that simvastatin is effective in inducing apoptosis in hyperproliferative pulmonary vascular lesions and could be considered as a potential drug for treatment of human SPH. PMID- 16698854 TI - Higher urine desmosine levels are associated with mortality in patients with acute lung injury. AB - Desmosine is a stable breakdown product of elastin that can be reliably measured in urine samples. We tested the hypothesis that higher baseline urine desmosine would be associated with higher mortality in 579 of 861 patients included in the recent Acute Respiratory Distress Syndrome Network trial of lower tidal volume ventilation (1). We also correlated urine desmosine levels with indexes of disease severity. Finally, we assessed whether urine desmosine was lower in patients who received lower tidal volumes. Desmosine was measured by radioimmunoassay in urine samples from days 0, 1, and 3 of the study. The data were expressed as a ratio of urine desmosine to urine creatinine to control for renal dilution. The results show that higher baseline (day 0) urine desmosine-to creatinine concentration was associated with a higher risk of death on adjusted analysis (odds ratio 1.36, 95% confidence interval 1.02-1.82, P=0.03). Urine desmosine increased in both ventilator groups from day 0 to day 3, but the average rise was higher in the 12-ml/kg predicted body weight group compared with the 6-ml/kg predicted body weight group (P=0.053, repeated-measures model). In conclusion, patients with acute lung injury ventilated with lower tidal volumes have lower urine desmosine levels, a finding that may reflect reduced extracellular matrix breakdown. These results illustrate the value of evaluating urinary biological markers that may have prognostic and pathogenetic significance in acute lung injury. PMID- 16698855 TI - Inhibition of matrix metalloproteinase-9 prevents neutrophilic inflammation in ventilator-induced lung injury. AB - Neutrophils are considered to play a central role in ventilator-induced lung injury (VILI). However, the pulmonary consequences of neutrophil accumulation have not been fully elucidated. Matrix metalloproteinase-9 (MMP-9) had been postulated to participate in neutrophil transmigration. The purpose of this study was to investigate the role of MMP-9 in the neutrophilic inflammation of VILI. Male Sprague-Dawley rats were divided into three groups: 1) low tidal volume (LVT), 7 ml/kg of tidal volume (VT); 2) high tidal volume (HVT), 30 ml/kg of VT; and 3) HVT with MMP inhibitor (HVT+MMPI). As a MMPI, CMT-3 was administered daily from 3 days before mechanical ventilation. Degree of VILI was assessed by wet-to dry weight ratio and acute lung injury (ALI) scores. Neutrophilic inflammation was determined from the neutrophil count in the lung tissue and myeloperoxidase (MPO) activity in the bronchoalveolar lavage fluid (BALF). MMP-9 expression and activity were examined by immunohistochemical staining and gelatinase zymography, respectively. The wet-to-dry weight ratio, ALI score, neutrophil infiltration, and MPO activity were increased significantly in the HVT group. However, in the HVT+MMPI group, pretreatment with MMPI decreased significantly the degree of VILI, as well as neutrophil infiltration and MPO activity. These changes correlated significantly with MMP-9 immunoreactivity and MMP-9 activity. Most outcomes were significantly worse in the HVT+MMPI group compared with the LVT group. In conclusion, VILI mediated by neutrophilic inflammation is closely related to MMP-9 expression and activity. The inhibition of MMP-9 protects against the development of VILI through the downregulation of neutrophil-mediated inflammation. PMID- 16698856 TI - Integrating acute lung injury and regulation of alveolar fluid clearance. AB - The acute respiratory distress syndrome (ARDS) is characterized by non cardiogenic pulmonary edema and flooding of the alveolar air spaces with proteinaceous fluid. ARDS develops in response to inflammatory stresses including sepsis, trauma, and severe pneumonia, and despite aggressive critical care management, it still has a mortality of 30-50%. At the time of its original description in 1967, relatively little was known about the specific mechanisms by which the alveolar epithelium regulated lung fluid balance. Over the last 20 years, substantial advances in our understanding of the alveolar epithelium have provided major new insights into how molecular and cellular mechanisms regulate the active transport of solutes and fluid across the alveolar epithelium under both normal and pathological conditions. Beginning with the elucidation of active sodium transport as a major driving force for the transport of water from the air space to the interstitium, elegant work by multiple investigators has revealed a complex and integrated network of membrane channels and pumps that coordinately regulates sodium, chloride, and water flux in both a cell- and condition-specific manner. At the Experimental Biology Meeting in San Francisco on April 4, 2006, a symposium was held to discuss some of the most recent advances. Although there is still much to learn about the mechanisms that impair normal alveolar fluid clearance under pathological conditions, the compelling experimental findings presented in this symposium raise the prospect that we are now poised to test and develop therapeutic strategies to improve outcome in patients with acute lung injury. PMID- 16698857 TI - Lung surfactant secretion by interalveolar Ca2+ signaling. AB - Although clusters of alveoli form the acinus, which is the most distal respiratory unit, it is not known whether interalveolar communication coordinates acinar surfactant secretion. To address this, we applied real-time digital imaging in conjunction with photo-excited Ca2+ uncaging in intact alveoli of the isolated, blood-perfused rat lung. We loaded alveolar cells with the Ca2+ cage o nitrophenyl EGTA-AM (NP-EGTA-AM) together with the fluorophores, fluo 4, or LysoTracker green (LTG) to determine, respectively, the cytosolic Ca2+ concentration ([Ca2+]cyt) or type 2 cell secretion. To uncage Ca2+ from NP-EGTA, we exposed a region in a selected alveolus to high-intensity UV illumination. As a result, fluo 4 fluorescence increased, whereas LTG fluorescence decreased, in the photo-targeted region, indicating that uncaging both increased [Ca2+]cyt and induced secretion. Concomitantly, [Ca2+]cyt increases conducted from the uncaging site induced type 2 cell secretion in both the selected alveolus as well as in neighboring alveoli, indicating the presence of interalveolar communication. These conducted responses were inhibited by pretreating alveoli with the connexin43 (Cx43)-inhibiting peptides gap 26 and gap 27. However, although the conducted [Ca2+]cyt increase diminished with distance from the uncaging site, type 2 cell secretion rates were similar at all locations. We conclude that Cx43 dependent, interalveolar Ca2+ signals regulate type 2 cell secretion in adjacent alveoli. Such interalveolar communication might facilitate acinar coordination of alveolar function. PMID- 16698858 TI - Alarms and human behaviour: implications for medical alarms. AB - Alarms are used in many clinical applications, but they are often less than optimal because the design and implementation of alarms has not always taken the cognitive capacity and processing mechanisms of the user into account. As a result alarms are frequently too loud, irritating, confusing, badly designed, and too numerous, resulting in them often being turned off and hindering, rather than enhancing, task performance. This paper reviews some of the main areas where it is essential to take account of the cognitive system of the user and behavioural processes more generally. Five central areas of concern are discussed: the number of alarms and ways that this might be reduced; false alarm rates and their impact on human responses; the design of alarms and the application of research into auditory cognition on design; intelligent alarm systems; and the proposals for alarm design set out in a recent worldwide medical alarms standard. In each area some background is given and the implications for alarm design and implementation outlined. The conclusion is that there are some indications that alarm design and implementation takes account of relevant research data, but that there is still some way to go before these findings are fully integrated and the situation is improved upon further. PMID- 16698860 TI - Monitoring the injured brain: ICP and CBF. AB - Raised intracranial pressure (ICP) and low cerebral blood flow (CBF) are associated with ischaemia and poor outcome after brain injury. Therefore, many management protocols target these parameters. This overview summarizes the technical aspects of ICP and CBF monitoring, and their role in the clinical management of brain-injured patients. Furthermore, some applications of these methods in current research are highlighted. ICP is typically measured using probes that are inserted into one of the lateral ventricles or the brain parenchyma. Therapeutic measures used to control ICP have relevant side-effects and continuous monitoring is essential to guide such therapies. ICP is also required to calculate cerebral perfusion pressure which is one of the most important therapeutic targets in brain-injured patients. Several bedside CBF monitoring devices are available. However, most do not measure CBF but rather a parameter that is thought to be proportional to CBF. Frequently used methods include transcranial Doppler which measures blood flow velocity and may be helpful for the diagnosis and monitoring of cerebral vasospasm after subarachnoid haemorrhage or jugular bulb oximetry which gives information on adequacy of CBF in relation to the metabolic demand of the brain. However, there is no clear evidence that incorporating data from CBF monitors into our management strategies improves outcome in brain-injured patients. PMID- 16698859 TI - Blood glucose concentration profile after 10 mg dexamethasone in non-diabetic and type 2 diabetic patients undergoing abdominal surgery. AB - BACKGROUND: Dexamethasone prevents postoperative nausea and vomiting but may increase blood glucose. We compared blood glucose concentrations after dexamethasone in non-diabetic and type 2 diabetic patients undergoing surgery and looked for any association with preoperative glycosylated haemoglobin [HbA (1c)] and BMI. METHODS: Sixty three patients were enrolled: 32 were non-diabetic (Group ND) and 31 type 2 diabetic (Group D) without insulin treatment. Anaesthesia was induced using i.v. anaesthetic agents and maintained with sevoflurane. All patients received 10 mg dexamethasone at induction. Blood glucose concentrations were measured at induction and then every 60 min for 240 min. Data were analysed using anova. Effects of HbA (1c) and BMI were investigated using linear correlation and logistic regression. RESULTS: Blood glucose concentrations increased significantly over time and peaked at 120 min after 10 mg dexamethasone in both groups. The magnitude of increase was comparable between the groups [mean (SD) 29 (19) and 35 (19)% of baseline in Group D and Group ND, respectively]. Maximum concentrations were higher in Group D [8.97 (1.51) mmol litre(-1), range 6.67-12.94 mmol litre(-1)] than in Group ND [7.86 (1.00) mmol litre(-1), range 5.78-10.00 mmol litre(-1)]. There was a significant correlation between the maximum concentrations and BMI (R(2)=0.21) or HbA (1c) (R(2)=0.26). Logistic regression analysis revealed that the higher the BMI, the lower the HbA (1c) threshold associated with an increased probability (>0.5) of observing blood glucose levels higher than 8.33 mmol litre(-1) during 240 min after dexamethasone administration. Similarly, the higher the HbA (1c), the lower the BMI threshold associated with the same probability. CONCLUSIONS: After 10 mg dexamethasone, blood glucose levels increase in non-diabetic and type 2 diabetic patients undergoing abdominal surgery. Poorly controlled diabetes and severe obesity can influence the development of hyperglycaemia. PMID- 16698861 TI - Cerebral microdialysis: research technique or clinical tool. AB - Cerebral microdialysis is a well-established laboratory tool that is increasingly used as a bedside monitor to provide on-line analysis of brain tissue biochemistry during neurointensive care. This review describes the principles of cerebral microdialysis and the rationale for its use in the clinical setting, including discussion of the most commonly used microdialysis biomarkers of acute brain injury. Potential clinical applications are reviewed and future research applications identified. Microdialysis has the potential to become an established part of mainstream multi-modality monitoring during the management of acute brain injury but at present should be considered a research tool for use in specialist centres. PMID- 16698862 TI - Influence on platelet aggregation of i.v. parecoxib and acetaminophen in healthy volunteers. AB - BACKGROUND: Acetaminophen (paracetamol) alone or in combination with other analgesics is widely used for postoperative analgesia. While acetaminophen and non-steroidal anti-inflammatory drugs inhibit platelet function, the cyclooxygenase-2 (COX-2) selectively inhibiting coxibs show no interference with platelet function. The authors studied the effect of a combination of i.v. parecoxib and acetaminophen on platelet function in healthy volunteers. METHODS: Eighteen healthy, male volunteers (22-33 yr) received i.v. acetaminophen 1 g, parecoxib 40 mg+acetaminophen 1 g or placebo in a double-blind, crossover study. Platelet function was assessed by photometric aggregometry and by measuring the release of thromboxane B(2). Plasma acetaminophen concentrations were measured by high-performance liquid chromatography. RESULTS: Platelet aggregation (median area under the curve) triggered with arachidonic acid 500 microM was 24.6, 3.9 and 4.2x10(3) area units (P=0.02, all groups) after placebo, acetaminophen and parecoxib+acetaminophen, respectively. Inhibition of platelet aggregation showed no difference between acetaminophen alone and the combination (P=0.82). Aggregation triggered with arachidonic acid 750 or 1000 microM, adenosine diphosphate (ADP) 1.5 or 3 microM, or epinephrine 5 microM showed no differences between the groups. Release of thromboxane B(2) in response to ADP was inhibited similarly by both acetaminophen and the combination. Plasma acetaminophen concentrations were similar after acetaminophen and the combination. CONCLUSIONS: Acetaminophen and parecoxib showed no interaction in inhibiting platelet function. In combination they cause a mild degree of COX-1 inhibition corresponding to that of acetaminophen alone. PMID- 16698863 TI - Clinical trials of monitoring in anaesthesia, critical care and acute ward care: a review. AB - During anaesthesia monitoring is used as part of a complex feedback-control system to keep the patient in a safe physiological 'envelope' and so is central to the conduct of a modern anaesthetic. The utility of basic monitoring is universally acknowledged and will never be assessed using randomized controlled trials. However, each time a new monitoring device is introduced, it can be assessed to see if it adds to the safety and effectiveness of anaesthetics. This review highlights some of the studies that have assessed new monitors in anaesthesia, critical care, and other areas of acute care. PMID- 16698864 TI - Anaesthetic management of a parturient with the postural orthostatic tachycardia syndrome: a case report. AB - Postural orthostatic tachycardia syndrome encompasses a group of disorders characterized by orthostatic intolerance. We describe the anaesthetic management of analgesia for labour and of Caesarean section in a parturient suffering from this disorder. Worsening of her symptoms during pregnancy was managed with an increase in the dose of beta-blockers taken by the patient. Epidural analgesia was instigated early to attenuate the stress of labour and avoid consequent triggering of a tachycardic response. Slow titration of epidural analgesia and anaesthesia after an adequate fluid preload was undertaken to minimize hypotension and subsequent tachycardia. Neuraxial opioid, combined with non steroidal anti-inflammatory drugs and bilateral iliohypogastric and ilioinguinal nerve blocks were used to optimize postoperative analgesia. PMID- 16698865 TI - Ultrasound-guided blocks of the ilioinguinal and iliohypogastric nerve: accuracy of a selective new technique confirmed by anatomical dissection. AB - BACKGROUND: Ilioinguinal and iliohypogastric nerve blocks may be used in the diagnosis of chronic groin pain or for analgesia for hernia repair. This study describes a new ultrasound-guided approach to these nerves and determines its accuracy using anatomical dissection control. METHODS: After having tested the new method in a pilot cadaver, 10 additional embalmed cadavers were used to perform 37 ultrasound-guided blocks of the ilioinguinal and iliohypogastric nerve. After injection of 0.1 ml of dye the cadavers were dissected to evaluate needle position and colouring of the nerves. RESULTS: Thirty-three of the thirty seven needle tips were located at the exact target point, in or directly at the ilioinguinal or iliohypogastric nerve. In all these cases the targeted nerve was coloured entirely. In two of the remaining four cases parts of the nerves were coloured. This corresponds to a simulated block success rate of 95%. In contrast to the standard 'blind' techniques of inguinal nerve blocks we visualized and targeted the nerves 5 cm cranial and posterior to the anterior superior iliac spine. The median diameters of the nerves measured by ultrasound were: ilioinguinal 3.0x1.6 mm, and iliohypogastric 2.9x1.6 mm. The median distance of the ilioinguinal nerve to the iliac bone was 6.0 mm and the distance between the two nerves was 10.4 mm. CONCLUSIONS: The anatomical dissections confirmed that our new ultrasound-guided approach to the ilioinguinal and iliohypogastric nerve is accurate. Ultrasound could become an attractive alternative to the 'blind' standard techniques of ilioinguinal and iliohypogastric nerve block in pain medicine and anaesthetic practice. PMID- 16698866 TI - Coronary stents and perioperative anti-platelet regimen: dilemma of bleeding and stent thrombosis. PMID- 16698867 TI - Adverse events in anaesthetic practice: qualitative study of definition, discussion and reporting. AB - BACKGROUND: This study aimed to explore how critical and acceptable practice are defined in anaesthesia and how this influences the discussion and reporting of adverse incidents. Method. We conducted workplace observations of, and interviews with, anaesthetists and anaesthetic staff. Transcripts were analysed qualitatively for recurrent themes and quantitatively for adverse events in anaesthetic process witnessed. We also observed departmental audit meetings and analysed meeting minutes and report forms. RESULTS: The educational value of discussing events was well-recognized; 28 events were discussed at departmental meetings, of which 5 (18%) were presented as 'critical incidents'. However, only one incident was reported formally. Our observations of anaesthetic practice revealed 103 minor events during the course of over 50 anaesthetic procedures, but none were acknowledged as offering the potential to improve safety, although some were direct violations of 'acceptable' practice. Formal reporting appears to be constrained by changing boundaries of what might be considered 'critical', by concerns of loss of control over formally reported incidents and by the perception that reporting schemes outside anaesthesia have purposes other than education. CONCLUSIONS: Despite clear official definitions of criticality in anaesthesia, there is ambiguity in how these are applied in practice. Many educationally useful events fall outside critical incident reporting schemes. Professional expertise in anaesthesia brings its own implicit safety culture but the reluctance to adopt a more explicit 'systems approach' to adverse events may impede further gains in patient safety in anaesthesia. PMID- 16698868 TI - Perioperative anaphylaxis from locally applied rifamycin SV and latex. AB - A patient developed severe anaphylaxis during irrigation of a wound with rifamycin SV. The temporal relationship between application of rifamycin SV, the positive skin test and basophil activation test for rifamycin SV strongly supported diagnosis of anaphylaxis from the locally applied antibiotic. However, after operation the patient had two anaphylactic reactions with pruritus, urticaria and angio-oedema after routine care by a nurse, and these were probably caused by natural rubber latex. This case report has several messages. First, it is not widely appreciated that topically applied drugs and related compounds can elicit life-threatening anaphylaxis. Second, it illustrates patients can present with more than one allergy. Finally, it provides an opportunity to summarize the applications of flow cytometry-assisted quantification of in vitro activated basophils in diagnosing the cause of anaphylaxis during anaesthesia. PMID- 16698869 TI - The Narcotrend 'depth of anaesthesia' monitor cannot reliably detect consciousness during general anaesthesia: an investigation using the isolated forearm technique. PMID- 16698870 TI - Comments on a case of left ventricular hypertrabeculation/noncompaction. PMID- 16698871 TI - The management of laryngeal and subglottic stenosis. PMID- 16698872 TI - Ephedrine and phenylephrine for treatment of maternal hypotension. PMID- 16698873 TI - Preparedness of intensive care units in south-east UK for influenza pandemic. PMID- 16698874 TI - Bispectral index monitoring for more effective electroconvulsive therapy? PMID- 16698876 TI - Glucose metabolism and acute myocardial infarction. PMID- 16698877 TI - Three-year duration of benefit from abciximab in patient receiving stents for acute myocardial infarction in the randomized double-blind ADMIRAL study. PMID- 16698878 TI - Dyspnoea after AZD6140: safety first? PMID- 16698879 TI - Suppression of Parkin enhances nigrostriatal and motor neuron lesion in mice over expressing human-mutated tau protein. AB - Abnormal deposition of protein tau takes place in the brain of patients with several neurodegenerative diseases. Few of these patients present frontotemporal dementia with parkinsonism and amyotrophy (FTDPA-17), an autosomal dominant tauopathy related to mutations of the gene that codes for protein tau, localized in chromosome 17. The great majority of patients with tauopathies such as Alzheimer's disease, sporadic frontotemporal dementia or progressive supranuclear palsy do not show a Mendelian pattern of inheritance. We have occasionally seen tauopathies in patients with parkin mutations and, therefore, hypothesized that the protein tau interacts with parkin. We have tested that hypothesis in mice with combined genetic modifications of tau (over-expression of human tau with three mutations known to produce FTDPA-17) and parkin (deleted) proteins. Homozygote parkin null or over-expressing mutated-human tau mice have subtle behavioral and molecular abnormalities but do not express a clinical phenotype of neurodegenerative disease. Mice with combined homozygous mutations of these two genes show progressively abnormal walking already noticeable at 3 months of age, loss of dopamine and dopamine markers in striatum, nuclear tau immunoreactive deposits in motor neurons of the spinal cord, abnormal expression of glial markers and enhanced levels of pro-apoptotic proteins; findings that were absent or less pronounced in homozygote animals with deletions of parkin or over expression of tau. The double transgenic mice do not express normal mechanisms of adaptation to stress such as increased levels of GSH and Hsp-70. In addition, they have reduced levels of CHIP-Hsc70, a complex known to attenuate aggregation of tau and to enhance ubiquitination of phosphorylated tau. We have found high levels of phosphorylated tau in parkin-/-+tau(VLW) mice and a relative decrease of the inactivated pSer9 to total GSK-3 levels. Our data reveal that there are interactions between tau and parkin that could be relevant for the pathogenesis and treatment of tauopathies. Similarly, we hope that the double transgenic parkin-/-+tau(VLW) mice could be useful for testing of compounds with putative therapeutic value in human tauopathies. PMID- 16698880 TI - Frequent genetic and epigenetic abnormalities contribute to the deregulation of cytoglobin in non-small cell lung cancer. AB - Lung cancer demonstrates the highest mortality in the UK. Previous studies have implicated allelic loss at chromosome 17q in the development of non-small cell lung carcinoma (NSCLC), and a number of known and putative tumour-suppressor genes reside within this region. One candidate tumour-suppressor gene is cytoglobin (CYGB), which is contained entirely within the 42.5 kb tylosis with oesophageal cancer (TOC) minimal region. CYGB abnormalities have been demonstrated only in sporadic head and neck cancers. In this study, we investigated the expression, promoter methylation and allelic imbalance status of this gene in 52 paired (normal/tumour) surgically excised lung tissue samples from patients with NSCLC. CYGB expression in tumour tissue was significantly reduced compared with corresponding adjacent normal in 54% of the examined cases (paired t-test, P<0.001). The CYGB promoter was shown by pyrosequencing to be significantly hypermethylated [2-fold increase of methylation index (MtI) in tumours] in 25/52 (48%) tumour samples compared with normal samples. MtI of the CYGB promoter was associated with CYGB mRNA expression (linear regression analysis, P=0.009), suggesting a primary role for the epigenetic events in CYGB silencing. In addition, frequent LOH was detected at the locus 17q25 in 32/48 (67%) tumours examined. It is of note that the loss of expression intensified when both LOH and hypermethylation coincided in samples (Mann-Whitney, P=0.049). These findings provide the first evidence to suggest the implication of CYGB in the pathogenesis of NSCLCs. PMID- 16698882 TI - Learning from policy failure and failing to learn from policy. PMID- 16698883 TI - Learning from policy failure? Health action zones in England. PMID- 16698884 TI - Cardiovascular risk factors in the Federation of Bosnia and Herzegovina. AB - BACKGROUND: Federation of Bosnia and Herzegovina (FBiH) has high cardiovascular disease mortality as other countries in Eastern Europe and situation has even deteriorated in the post war period. Reliable information on risk factor levels and patterns needed in prevention planning and disease management has been lacking. METHODS: A cross sectional population survey was conducted in the FBiH in autumn 2002. A random sample of population, aged 25-64 years, was taken using a three stage stratified sampling methodology. Altogether, 2750 persons participated in the survey (1121 men and 1629 women). The survey was done according to internationally established standards and protocols. RESULTS: The mean systolic blood pressure was 132 mmHg among men and 135 mmHg among women. The mean diastolic blood pressure was 84 mmHg in both genders. Almost 40% of the participants were recognized as hypertensive (blood pressure level over 140/90 mmHg). The prevalence of hypertension among men was 36% and among women 45%. The mean Body Mass Index (BMI) was 26.5 kg/m(2) among males and 27.0 kg/m(2) among females. About 75% of both men and women were overweight (BMI > 25 kg/m(2)) and 16% of men and 20% of women were obese (BMI > 30 kg/m(2)). About 50% of men and 30% of women reported to be daily smokers. CONCLUSIONS: As a whole the non communicable disease risk factor levels in the FBiH seems to be relatively high. The data can be utilized in health promotion planning and as a baseline for future monitoring activities with possibility of international comparison of results. PMID- 16698885 TI - Can pigeons change the emergency system in Poland? PMID- 16698886 TI - AIDS mortality before and after the introduction of highly active antiretroviral therapy: does it vary with socioeconomic group in a country with a National Health System? AB - BACKGROUND: The aim of this study is to determine whether socioeconomic AIDS mortality inequalities before and after the introduction of highly active antiretroviral therapy (HAART) have increased or decreased in a Spanish city where HAART is free. METHODS: The study used a trend design, including all Barcelona residents older than 19 years of age. All AIDS deaths, which occurred among these residents between 1991 and 2001 were included. The variables studied were age, sex, socioeconomic (SES) group and HIV transmission group. AIDS age standardized mortality rates for each year were estimated. Poisson regression models were fitted to obtain the relative risk (RR) of AIDS death for each socioeconomic group with respect to the reference group. RESULTS: AIDS mortality increased up until 1995 and subsequently decreased due to the introduction of HAART. The increase in AIDS mortality was greater in the lowest SES group, which had higher rates and a RR of dying larger than that of the highest SES group, fact that remained fairly stable over the whole period. A similar pattern was observed in intravenous drug users. In the homosexual transmission group, rates for the lowest SES group were higher for the whole period and increased until 1996, while rates for the other SES groups were lower and decreased over the entire period. CONCLUSIONS: The fact that inequalities in AIDS mortality by SES group remained fairly stable for the whole period suggests that perhaps access to HAART, or adherence, is lower than desirable, in people of lower SES groups. These results ought to be taken into account when implementing treatment and prevention strategies. PMID- 16698887 TI - Adolescent obesity in Lebanese private schools. AB - BACKGROUND: Obesity has become a public health problem worldwide. Our objective was to calculate the prevalence of overweight and obesity. METHODS: It is a cross sectional study of adolescents in private Lebanese schools, aged 10-18 years. Gender, birth date and measures of weight and height were recorded. RESULTS: In 12,299 adolescents, we found high prevalence of obesity (7.5%) and at risk of obesity (24.4%). In girls, risk of obesity and obesity prevalence decrease with increasing age (P < 10(-4)) as compared with that in boys. CONCLUSION: Early recognition of obesity should become routine in paediatric ambulatory care settings. PMID- 16698888 TI - Nuclear DNA content variation among Central European Koeleria taxa. AB - BACKGROUND AND AIMS: Polyploidization plays an important role in the evolution of many plant genera, including Koeleria. The knowledge of ploidy, chromosome number and genome size may enable correct taxonomic treatment when other features are insufficient as in Koeleria. Therefore, these characteristics and their variability were determined for populations of six central European Koeleria taxa. METHODS: Chromosome number analysis was performed by squashing root meristems, and ploidy and 2C nuclear DNA content were estimated by flow cytometry. KEY RESULTS: Three diploids (K. glauca, K. macrantha var. macrantha and var. pseudoglauca), one tetraploid (K. macrantha var. majoriflora), one decaploid (K. pyramidata) and one dodecaploid (K. tristis) were found. The 2C nuclear DNA content of the diploids ranged from 4.85 to 5.20 pg. The 2C DNA contents of tetraploid, decaploid and dodecaploid taxa were 9.31 pg, 22.89 pg and 29.23 pg, respectively. The DNA content of polyploids within the K. macrantha aggregate (i.e. within K. macrantha and K. pyramidata) was smaller than the expected multiple of the diploid genome (K. macrantha var. macrantha). Geography correlated variation of DNA content was found for some taxa. Czech populations of K. macrantha var. majoriflora had a 5.06% smaller genome than the Slovak ones. An isolated eastern Slovakian population of K. tristis revealed 8.04% less DNA than populations from central Slovakia. In central and north-west Bohemia, diploid and tetraploid cytotypes of K. macrantha were sympatric; east from this region diploid populations, and towards the west tetraploid populations were dominant. CONCLUSIONS: Remarkable intra-specific inter-population differences in nuclear DNA content were found between Bohemian and Pannonian populations of Koeleria macrantha var. majoriflora and between geographically isolated central and eastern Slovakian populations of K. tristis. These differences occur over a relatively small geographical scale. PMID- 16698889 TI - Nicorandil-associated anal ulceration. PMID- 16698890 TI - In vitro and in vivo correlation of hepatic transporter effects on erythromycin metabolism: characterizing the importance of transporter-enzyme interplay. AB - The effects of hepatic uptake and efflux transporters on erythromycin (ERY) disposition and metabolism were examined by comparing results from rat hepatic microsomes, freshly isolated hepatocytes, and in vivo studies. Uptake studies carried out in freshly isolated rat hepatocytes showed that ERY and its metabolite (N-demethyl-ERY) are substrates of Oatp1a4 and Oatp1b2. Whereas rifampin and GG918 [GF120918: N-{4-[2-(1,2,3,4-tetrahydro-6,7-dimethoxy-2 isoquinolinyl)-ethyl]-phenyl}-9,10-dihydro-5-methoxy-9-oxo-4-acridine carboxamine] exerted minimal effects on metabolism in microsomes, rifampin (2.5 microM) and GG918 (0.5 microM) significantly decreased and increased ERY metabolism in hepatocytes, respectively. Concentration-time course studies further demonstrated that, compared with the intracellular N-demethyl-ERY control area under the curve (AUC) (0.795 +/- 0.057 microM . min), a decreased AUC (0.513 +/- 0.028 microM . min, p < 0.005) was observed when ERY was coincubated with rifampin, and an increased AUC (2.14 +/- 0.21 microM . min, p < 0.05) was found when GG918 was present. The results of the i.v. bolus studies showed that, compared with the ERY clearance of the controls (47.2 +/- 12.5 ml/min/kg for the rifampin group and 42.1 +/- 5.7 for the GG918 group), a decreased blood clearance, 29.8 +/- 6.1 ml/min/kg (p < 0.05) and 21.7 +/- 9.0 ml/min/kg (p < 0.01), was observed when rifampin or GG918, respectively, was coadministered. When either inhibitor was codosed with ERY, volume of distribution at steady state was unchanged, but t1/2 and mean residence time significantly increased compared with the controls. Hepatic uptake and efflux transporters modulate intracellular concentrations of ERY, thereby affecting metabolism. The interplay of transporters and enzymes must be considered in evaluating potential drug-drug interactions. PMID- 16698891 TI - In silico prediction of drug binding to CYP2D6: identification of a new metabolite of metoclopramide. AB - Patients with cancer often take many different classes of drugs to treat the effects of their malignancy and the side effects of treatment, as well as their comorbidities. The potential for drug-drug interactions that may affect the efficacy of anticancer treatment is high, and a major source of such interactions is competition for the drug-metabolizing enzymes, cytochromes P450 (P450s). We have examined a series of 20 drugs commonly prescribed to cancer patients to look for potential interactions via CYP2D6. We used a homology model of CYP2D6, together with molecular docking techniques, to perform an in silico screen for binding to CYP2D6. Experimental IC50 values were determined for these compounds and compared with the model predictions to reveal a correlation with a regression coefficient of r2= 0.61. Importantly, the docked conformation of the commonly prescribed antiemetic metoclopramide predicted a new site of metabolism that was further investigated through in vitro analysis with recombinant CYP2D6. An aromatic N-hydroxy metabolite of metoclopramide, consistent with predictions from our modeling studies, was identified by high-performance liquid chromatography/mass spectrometry. This metabolite was found to represent a major product of metabolism in human liver microsomes, and CYP2D6 was identified as the main P450 isoform responsible for catalyzing its formation. In view of the prevalence of interindividual variation in the CYP2D6 genotype and phenotype, we suggest that those experiencing adverse reactions with metoclopramide, e.g., extrapyramidal syndrome, are likely to have a particular CYP2D6 genotype/phenotype. This warrants further investigation. PMID- 16698892 TI - Species differences in metabolism and pharmacokinetics of a sphingosine-1 phosphate receptor agonist in rats and dogs: formation of a unique glutathione adduct in the rat. AB - The pharmacokinetics and metabolism of 1-(4-((4-phenyl-5-trifluoromethyl-2 thienyl)methoxy)benzyl)azetidine-3-carboxylic acid (MRL-A), a selective agonist for the sphingosine-1-phosphate 1 (S1P1) receptor, were investigated in rats and dogs. In both species, more than 50% of the dose was excreted in bile. Specific to the rat, and observed in bile, were a taurine conjugate of MRL-A and a glucuronide conjugate of an azetidine lactam metabolite. In dogs, a smaller portion of the dose (54% of administered dose) was excreted intact in bile, and the major metabolites detected were an azetidine N-oxide of MRL-A and an acylglucuronide of an N-dealkylation product. This latter metabolite was also observed in rat bile. Stereoselective formation of the N-oxide isomer was observed in dogs, whereas the rat produced comparable amounts of both isomers. The formation of a unique glutathione adduct was observed in rat bile, which was proposed to occur via N-dealkylation, followed by reduction of the putative aldehyde product to form the alcohol, and dehydration of the alcohol to generate a reactive quinone methide intermediate. Incubation of a synthetic standard of this alcohol in rat microsomes fortified with reduced glutathione or rat hepatocytes resulted in formation of this unique glutathione adduct. PMID- 16698893 TI - NMR characterization of an S-linked glucuronide metabolite of the potent, novel, nonsteroidal progesterone agonist tanaproget. AB - Tanaproget is a first-in-class nonsteroidal progesterone receptor agonist that is being investigated for use in contraception. A major in vitro and in vivo metabolite of tanaproget formed in humans was initially characterized as a glucuronide of tanaproget. However, whether the glucuronide was linked to the nitrogen or sulfur of the benzoxazine-2-thione group in tanaproget could not be determined by liquid chromatography/mass spectrometry (LC/MS) and LC-tandem mass spectrometry analysis. To obtain additional structural details for this metabolite, additional quantities were generated from rat liver microsomal incubations and purified by high-performance liquid chromatography (HPLC) for NMR analysis. The NMR data for the metabolite confirmed that the glucuronide was covalently bound to either the sulfur or the nitrogen of the benzoxazine-2-thione moiety. The lack of key through-bond (scalar) and through-space (dipolar) one dimensional (1D) and two-dimensional (2D) NMR couplings and correlations in the metabolite spectra (due primarily to low sample concentration) precluded an unambiguous structure elucidation. Subsequent synthesis of the S- and N glucuronides of tanaproget from tanaproget facilitated the unambiguous regio- and stereochemical assignment of the metabolite by comparison of 1D NMR chemical shifts and scalar coupling constants, 2D NMR correlations, and HPLC and LC/MS characteristics between the synthetic compounds and the metabolite. From extensive comparison of the spectral and chromatographic data of the microsomally derived metabolite and the synthetic compounds, the metabolite has been determined to be the S-(beta)-D-glucuronide of tanaproget. PMID- 16698894 TI - The role of supraspinal vasopressin and glutamate neurones in an increase in renal sympathetic activity in response to mild haemorrhage in the rat. AB - This study investigated the importance of supraspinal vasopressin and glutamate neurones in regulating renal sympathetic activity as part of the response to an acute reduction in blood volume. Wistar rats anaesthetized with chloralose and urethane were instrumented to record arterial blood pressure, heart rate and left renal sympathetic nerve activity. Pharmacological agonists and antagonists to glutamate and vasopressin were applied to the renal outflow of the spinal cord via an intrathecal catheter inserted at the foramen magnum and with the tip at the level of T10. Both glutamate and vasopressin increased renal sympathetic activity, and these actions were shown to be selectively blocked by their respective antagonists. Removing 1 ml of venous blood from a femoral venous catheter elicited an increase of 26 +/- 2% in renal sympathetic activity. This response to mild haemorrhage was halved to 13 +/- 4% by prior intrathecal application of a selective V1a antagonist. Similarly, prior intrathecal application of kynurenic acid reduced the response to the mild haemorrhage from 28 +/- 2 to 12.6 +/- 2.8%. Intrathecal application of both antagonists together reduced the haemorrhage response even further to 8 +/- 3%. All the changes were statistically significant at P < 0.01. It is concluded that a small reduction in blood volume induces an increase in renal sympathetic activity dependent on vasopressin and glutamate release from terminals of supraspinal neurones. It is suggested that the vasopressin neurones most probably originate from the paraventricular nucleus of the hypothalamus. PMID- 16698895 TI - Early steps in the biosynthesis of NAD in Arabidopsis start with aspartate and occur in the plastid. AB - NAD is a ubiquitous coenzyme involved in oxidation-reduction reactions and is synthesized by way of quinolinate. Animals and some bacteria synthesize quinolinate from tryptophan, whereas other bacteria synthesize quinolinate from aspartate (Asp) using L-Asp oxidase and quinolinate synthase. We show here that Arabidopsis (Arabidopsis thaliana) uses the Asp-to-quinolinate pathway. The Arabidopsis L-Asp oxidase or quinolinate synthase gene complemented the Escherichia coli mutant defective in the corresponding gene, and T-DNA-based disruption of either of these genes, as well as of the gene coding for the enzyme quinolinate phosphoribosyltransferase, was embryo lethal. An analysis of functional green fluorescent protein-fused constructs and in vitro assays of uptake into isolated chloroplasts demonstrated that these three enzymes are located in the plastid. PMID- 16698896 TI - Carbon balance and circadian regulation of hydrolytic and phosphorolytic breakdown of transitory starch. AB - Transitory starch is formed in chloroplasts during the day and broken down at night. Transitory starch degradation could be regulated by light, circadian rhythms, or carbon balance. To test the role of these potential regulators, starch breakdown rates and metabolites were measured in bean (Phaseolus vulgaris) and Arabidopsis (Arabidopsis thaliana) plants. In continuous light, starch and maltose levels oscillated in a circadian manner. Under photorespiratory conditions, transitory starch breakdown occurred in the light faster than at night and glucose-6-P (G6P) was elevated. Nonaqueous fractionation showed that the increase in G6P occurred in the chloroplast. When Arabidopsis plants lacking the plastidic starch phosphorylase enzyme were placed under photorespiratory conditions, G6P levels remained constant, indicating that the increased chloroplastic G6P resulted from phosphorolytic starch degradation. Maltose was increased under photorespiratory conditions in both wild type and plants lacking starch phosphorylase, indicating that regulation of starch breakdown may occur at a point preceding the division of the hydrolytic and phosphorolytic pathways. When bean leaves were held in N2 to suppress photosynthesis and Suc synthesis without increasing photorespiration, starch breakdown did not occur and maltose and G6P levels remained constant. The redox status of the chloroplasts was found to be oxidized under conditions favoring starch degradation. PMID- 16698897 TI - Surface position, not signaling from surrounding maternal tissues, specifies aleurone epidermal cell fate in maize. AB - Maize (Zea mays) endosperm consists of an epidermal-like surface layer of aleurone cells, an underlying body of starchy endosperm cells, and a basal layer of transfer cells. To determine whether surrounding maternal tissues perform a role in specifying endosperm cell fates, a maize endosperm organ culture technique was established whereby the developing endosperm is completely removed from surrounding maternal tissues. Using cell type-specific fluorescence markers, we show that aleurone cell fate specification occurs exclusively in response to surface position and does not require specific, continued maternal signal input. The starchy endosperm and aleurone cell fates are freely interchangeable throughout the lifespan of the endosperm, with internalized aleurone cells converting to starchy endosperm cells and with starchy endosperm cells that become positioned at the surface converting to aleurone cells. In contrast to aleurone and starchy endosperm cells, transfer cells fail to develop in in vitro grown endosperm, supporting earlier indications that maternal tissue interaction is required to fully differentiate this cell type. Several parameters confirm that the maize endosperm organ cultures described herein retain the main developmental features of in planta endosperm, including fidelity of aleurone mutant phenotypes, temporal and spatial control of cell type-specific fluorescent markers, specificity of cell type transcripts, and control of mitotic cell divisions. PMID- 16698898 TI - A suppressor of fab1 challenges hypotheses on the role of thylakoid unsaturation in photosynthetic function. AB - Leaf membrane lipids of the Arabidopsis (Arabidopsis thaliana) fatty acid biosynthesis 1 (fab1) mutant contain a 35% to 40% increase in the predominant saturated fatty acid 16:0, relative to wild type. This increase in membrane saturation is associated with loss of photosynthetic function and death of mutant plants at low temperatures. We have initiated a suppressor screen for mutations that allow survival of fab1 plants at 2 degrees C. Five suppressor mutants identified in this screen all rescued the collapse of photosynthetic function observed in fab1 plants. While fab1 plants died after 5 to 7 weeks at 2 degrees C, the suppressors remained viable after 16 weeks in the cold, as judged by their ability to resume growth following a return to 22 degrees C and to subsequently produce viable seed. Three of the suppressors had changes in leaf fatty acid composition when compared to fab1, indicating that one mechanism of suppression may involve compensating changes in thylakoid lipid composition. Surprisingly, the suppressor phenotype in one line, S31, was associated with a further substantial increase in lipid saturation. The overall leaf fatty acid composition of S31 plants contained 31% 16:0 compared with 23% in fab1 and 17% in wild type. Biochemical and genetic analysis showed that S31 plants contain a new allele of fatty acid desaturation 5 (fad5), fad5-2, and are therefore partially deficient in activity of the chloroplast 16:0 Delta7 desaturase. A double mutant produced by crossing fab1 to the original fad5-1 allele also remained alive at 2 degrees C, indicating that the fad5-2 mutation is the suppressor in the S31 (fab1 fad5-2) line. Based on the biophysical characteristics of saturated and unsaturated fatty acids, the increased 16:0 in fab1 fad5-2 plants would be expected to exacerbate, rather than ameliorate, low-temperature damage. We propose instead that a change in shape of the major thylakoid lipid, monogalactosyldiacylglycerol, mediated by the fad5-2 mutation, may compensate for changes in lipid structure resulting from the original fab1 mutation. Our identification of mutants that suppress the low temperature phenotype of fab1 provides new tools to understand the relationship between thylakoid lipid structure and photosynthetic function. PMID- 16698899 TI - Identification of a mung bean arabinofuranosyltransferase that transfers arabinofuranosyl residues onto (1, 5)-linked alpha-L-arabino-oligosaccharides. AB - Arabinofuranosyltransferase activity was identified in Golgi membranes obtained from mung bean (Vigna radiata) hypocotyls. The enzyme transfers the arabinofuranosyl (Araf) residue from UDP-beta-L-arabinofuranose to exogenous (1, 5)-linked alpha-L-arabino-oligosaccharides labeled at their reducing ends with 2 aminobenzamide. The transferred residue was shown, using 1H-nuclear magnetic resonance spectroscopy and alpha-L-arabinofuranosidase treatment, to be alpha-L Araf and to be linked to O-5 of the nonreducing terminal Araf residue of the acceptor oligosaccharide. The enzyme was nonprocessive because only a single Araf residue was added to the acceptor molecule. Arabino-oligosaccharides with a degree of polymerization between 3 and 8 were acceptor substrates. The 2 aminobenzamide-labeled arabino-tetra- and pentasaccharides were the most effective acceptor substrates analyzed. The enzyme has a pH optimum between 6.5 and 7.0 and its activity is stimulated by Mn2+ and Co2+ ions. The apparent Km and Vmax values of the arabinofuranosyltransferase for UDP-arabinofuranose are 243 microm and 243 pmol min(-1) mg protein(-1), respectively. The highest enzyme activity was detected in the elongating regions of mung bean hypocotyls. The data show that UDP-arabinofuranose is the donor molecule for the generation of arabino oligosaccharides composed of Araf residues. PMID- 16698900 TI - The SCABRA3 nuclear gene encodes the plastid RpoTp RNA polymerase, which is required for chloroplast biogenesis and mesophyll cell proliferation in Arabidopsis. AB - In many plant species, a subset of the genes of the chloroplast genome is transcribed by RpoTp, a nuclear-encoded plastid-targeted RNA polymerase. Here, we describe the positional cloning of the SCABRA3 (SCA3) gene, which was found to encode RpoTp in Arabidopsis (Arabidopsis thaliana). We studied one weak (sca3-1) and two strong (sca3-2 and sca3-3) alleles of the SCA3 gene, the latter two showing severely impaired plant growth and reduced pigmentation of the cotyledons, leaves, stem, and sepals, all of which were pale green. The leaf surface was extremely crumpled in the sca3 mutants, although epidermal cell size and morphology were not perturbed, whereas the mesophyll cells were less densely packed and more irregular in shape than in the wild type. A significant reduction in the size, morphology, and number of chloroplasts was observed in homozygous sca3-2 individuals whose photoautotrophic growth was consequently perturbed. Microarray analysis showed that several hundred nuclear genes were differentially expressed in sca3-2 and the wild type, about one-fourth of which encoded chloroplast-targeted proteins. Quantitative reverse transcription-PCR analyses showed that the sca3-2 mutation alters the expression of the rpoB, rpoC1, clpP, and accD plastid genes and the SCA3 paralogs RpoTm and RpoTmp, which respectively encode nuclear-encoded mitochondrion or dually targeted RNA polymerases. Double mutant analysis indicated that RpoTmp and SCA3 play redundant functions in plant development. Our findings support a role for plastids in leaf morphogenesis and indicate that RpoTp is required for mesophyll cell proliferation. PMID- 16698901 TI - NUCLEAR FUSION DEFECTIVE1 encodes the Arabidopsis RPL21M protein and is required for karyogamy during female gametophyte development and fertilization. AB - Karyogamy, or nuclear fusion, is essential for sexual reproduction. In angiosperms, karyogamy occurs three times: twice during double fertilization of the egg cell and the central cell and once during female gametophyte development when the two polar nuclei fuse to form the diploid central cell nucleus. The molecular mechanisms controlling karyogamy are poorly understood. We have identified nine female gametophyte mutants in Arabidopsis (Arabidopsis thaliana), nuclear fusion defective1 (nfd1) to nfd9, that are defective in fusion of the polar nuclei. In the nfd1 to nfd6 mutants, failure of fusion of the polar nuclei is the only defect detected during megagametogenesis. nfd1 is also affected in karyogamy during double fertilization. Using transmission electron microscopy, we showed that nfd1 nuclei fail to undergo fusion of the outer nuclear membranes. nfd1 contains a T-DNA insertion in RPL21M that is predicted to encode the mitochondrial 50S ribosomal subunit L21, and a wild-type copy of this gene rescues the mutant phenotype. Consistent with the predicted function of this gene, an NFD1-green fluorescent protein fusion protein localizes to mitochondria and the NFD1/RPL21M gene is expressed throughout the plant. The nfd3, nfd4, nfd5, and nfd6 mutants also contain T-DNA insertions in genes predicted to encode proteins that localize to mitochondria, suggesting a role for this organelle in nuclear fusion. PMID- 16698902 TI - Redox regulation of a novel plastid-targeted beta-amylase of Arabidopsis. AB - Nine genes of Arabidopsis (Arabidopsis thaliana) encode for beta-amylase isozymes. Six members of the family are predicted to be extrachloroplastic isozymes and three contain predicted plastid transit peptides. Among the latter, chloroplast-targeted beta-amylase (At4g17090) and thioredoxin-regulated beta amylase (TR-BAMY; At3g23920; this work) are experimentally demonstrated to be targeted to plastids. Recombinant TR-BAMY was catalytically active only when expressed as a mature protein, i.e. with no transit peptide. Mature TR-BAMY was a monomer of 60 kD, hydrolyzing soluble starch with optimal activity between pH 6.0 and 8.0. The activity of recombinant TR-BAMY was strictly dependent on redox potential with an Em,7.0 of -302 +/- 14 mV. Thioredoxins f1, m1, and y1 of Arabidopsis were all able to mediate the reductive activation of oxidized TR BAMY. Site-specific mutants showed that TR-BAMY oxidative inhibition depended on the formation of a disulfide bridge between Cys-32 and Cys-470. Consistent with TR-BAMY redox dependency, total beta-amylase activity in Arabidopsis chloroplasts was partially redox regulated and required reducing conditions for full activation. In Arabidopsis, TR-BAMY transcripts were detected in leaves, roots, flowers, pollen, and seeds. TR-BAMY may be the only beta-amylase of nonphotosynthetic plastids suggesting a redox regulation of starch metabolism in these organelles. In leaves, where chloroplast-targeted beta-amylase is involved in physiological degradation of starch in the dark, TR-BAMY is proposed to participate to a redox-regulated pathway of starch degradation under specific stress conditions. PMID- 16698903 TI - Determination of structural requirements and probable regulatory effectors for membrane association of maize sucrose synthase 1. AB - Sucrose (Suc) synthase (SUS) cleaves Suc to form UDP glucose and fructose, and exists in soluble and membrane-associated forms, with the latter proposed to channel UDP glucose to the cellulose-synthase complex on the plasma membrane of plant cells during synthesis of cellulose. However, the structural features responsible for membrane localization and the mechanisms regulating its dual intracellular localization are unknown. The maize (Zea mays) SUS1 isoform is likely to have the intrinsic ability to interact directly with membranes because we show: (1) partial membrane localization when expressed in Escherichia coli, and (2) binding to carbonate-stripped plant microsomes in vitro. We have undertaken mutational analyses (truncations and alanine substitutions) and in vitro microsome-binding assays with the SUS1 protein to define intrinsic membrane binding regions and potential regulatory factors that could be provided by cellular microenvironment. The results suggest that two regions of SUS1 contribute to membrane affinity: (1) the amino-terminal noncatalytic domain, and (2) a region with sequence similarity to the C-terminal pleckstrin homology domain of human pleckstrin. Alanine substitutions within the pleckstrin homology like domain of SUS1 reduced membrane association in E. coli and with plant microsomes in vitro without reducing enzymatic activity. Microsomal association of wild-type SUS1 displayed cooperativity with SUS1 protein concentration and was stimulated by both lowering the pH and adding Suc. These studies offer insight into the molecular level regulation of SUS1 localization and its participation in carbon partitioning in plants. Moreover, transgenics with active SUS mutants altered in membrane affinity may be of technological utility. PMID- 16698904 TI - Necrosis- and ethylene-inducing peptide from Fusarium oxysporum induces a complex cascade of transcripts associated with signal transduction and cell death in Arabidopsis. AB - Treatment of Arabidopsis (Arabidopsis thaliana) with a necrosis- and ethylene inducing peptide (Nep1) from Fusarium oxysporum inhibited both root and cotyledon growth and triggered cell death, thereby generating necrotic spots. Nep1-like proteins are produced by divergent microbes, many of which are plant pathogens. Nep1 in the plant was localized to the cell wall and cytosol based on immunolocalization results. The ratio of chlorophyll a fluorescence (F685 nm/F730 nm) significantly decreased after 75-min treatment with Nep1 in comparison to the control. This suggested that a short-term compensation of photosynthesis occurred in response to localized damage to cells. The concentrations of most water soluble metabolites analyzed were reduced in Arabidopsis seedlings after 6 h of Nep1 treatment, indicating that the integrity of cellular membranes had failed. Microarray results showed that short-term treatment with Nep1 altered expression of numerous genes encoding proteins putatively localized to organelles, especially the chloroplast and mitochondria. Short-term treatment with Nep1 induced multiple classes of genes involved in reactive oxygen species production, signal transduction, ethylene biosynthesis, membrane modification, apoptosis, and stress. Quantitative PCR was used to confirm the induction of genes localized in the chloroplast, mitochondria, and plasma membrane, and genes responsive to calcium/calmodulin complexes, ethylene, jasmonate, ethylene biosynthesis, WRKY, and cell death. The majority of Nep1-induced genes has been associated with general stress responses but has not been critically linked to resistance to plant disease. These results are consistent with Nep1 facilitating cell death as a component of diseases caused by necrotrophic plant pathogens. PMID- 16698907 TI - Evidence is a challenge in wound management. PMID- 16698905 TI - Heavy metal stress and sulfate uptake in maize roots. AB - ZmST1;1, a putative high-affinity sulfate transporter gene expressed in maize (Zea mays) roots, was functionally characterized and its expression patterns were analyzed in roots of plants exposed to different heavy metals (Cd, Zn, and Cu) interfering with thiol metabolism. The ZmST1;1 cDNA was expressed in the yeast (Saccharomyces cerevisiae) sulfate transporter mutant CP154-7A. Kinetic analysis of sulfate uptake isotherm, determined on complemented yeast cells, revealed that ZmST1;1 has a high affinity for sulfate (Km value of 14.6 +/- 0.4 microm). Cd, Zn, and Cu exposure increased both ZmST1;1 expression and root sulfate uptake capacity. The metal-induced sulfate uptakes were accompanied by deep alterations in both thiol metabolism and levels of compounds such as reduced glutathione (GSH), probably involved as signals in sulfate uptake modulation. Cd and Zn exposure strongly increased the level of nonprotein thiols of the roots, indicating the induction of additional sinks for reduced sulfur, but differently affected root GSH contents that decreased or increased following Cd or Zn stress, respectively. Moreover, during Cd stress a clear relation between the ZmST1;1 mRNA abundance increment and the entity of the GSH decrement was impossible to evince. Conversely, Cu stress did not affect nonprotein thiol levels, but resulted in a deep contraction of GSH pools. Our data suggest that during heavy metal stress sulfate uptake by roots may be controlled by both GSH-dependent or independent signaling pathways. Finally, some evidence suggesting that root sulfate availability in Cd-stressed plants may limit GSH biosynthesis and thus Cd tolerance are discussed. PMID- 16698908 TI - Microangiopathic ulcers of the lower extremities: a daily challenge. PMID- 16698909 TI - Prognostic factors of leg amputation in patients with vascular injury: a systematic review. AB - Individual studies on the prognostic factors of leg amputation, due to vascular injury, have been small, and they have produced conflicting results. Reliable data are necessary so that surgery can be targeted more effectively. The authors carried out a systematic review from 1990 to 2002 to identify the high risk of patients to amputation. Meta-analysis was carried out. The authors found that patients with preoperative hypotension, popliteal artery injury, and associated bone and nerve injury had a significantly higher risk of leg amputation than those without these risk factors. Also, patients with postoperative infection had a higher chance of amputation than those without infection. This information is essential for an appropriate evaluation and the treatment of such patients. PMID- 16698910 TI - A study of the prevalence and risk factors of foot problems in a population of diabetic patients in cameroon. AB - Foot problems are common in diabetic patients and are one of the most expensive chronic complications to treat. The authors sought to determine the prevalence and risk factors of the diabetic foot in a clinic population. In this cross sectional study of 300 diabetic patients, the authors reviewed records, carried out an interview, and performed a meticulous foot examination with assessment of neuropathy (monofilaments and tuning fork) and ischemia (pulses). Foot lesions were classified according to Wagner grades. The prevalence of foot lesions was 13.0% (inpatients 25.6% and outpatients 11.1%). Diabetic neuropathy assessed using monofilaments was found in 81 patients (27.3%) (monofilaments). The prevalence of ischemia was 21.3% and deformity was 17.3%, whereas 37 patients (12.3%) had a previous history of foot lesions. Foot examination was done in 14.3% of patients, and 47% had a risky nail-trimming habit, whereas 22% wore ill fitting shoes. The prevalence of diabetic foot lesions is high, and known risk factors are significantly present, especially poor foot care. PMID- 16698911 TI - Diagnostic criteria and treatment of Buerger's disease: a review. AB - Buerger's disease is an inflammatory occlusive disorder affecting the small and medium-size arteries and veins of young, predominantly male, smokers. The disorder has been identified as an autoimmune response triggered when nicotine is present. Tobacco abuse is the major contributing risk factor; however, smoking seems to be a synergistic factor rather than the cause of the disease. The traditional diagnosis of Buerger's disease is based on 5 criteria (smoking history, onset before the age of 50 years, infrapopliteal arterial occlusive disease, either upper limb involvement or phlebitis migrans, and absence of atherosclerotic risk factors other than smoking). As there is no specific diagnostic test and an absence of positive serologic markers, confident clinical diagnosis should be made only when all these 5 criteria have been fulfilled although not universally accepted. The angiographic findings in Buerger's disease ("corkscrew," "spider legs," or "tree roots") are helpful but not pathognomonic. A wide spectrum of medical or surgical therapeutic options have been proposed; however, total abstinence from tobacco use remains the only means of stopping the disease progression. The initial management of patients with Buerger's disease should be conservative. Because several arteries may be unaffected, claudicants should be encouraged to walk, whereas patients with "critical" ischemia should be admitted for bed rest in the hospital. Bypass grafting is seldom an option, as the location of the lesions distally leaves little to bypass because of lack of target vessels. A literature review revealed only a few series reporting vascular reconstruction (mainly femorodistal bypasses) in Buerger's disease. Bypass patency rates were suboptimal; however, the corresponding limb salvage rates were satisfactory. A possible explanation is that patent grafts, even over a short period of time, are sufficient to allow healing of ulcers in patients with Buerger's disease. PMID- 16698912 TI - A unique waterskiing injury leading to a necrotizing foot infection in an insulin dependent diabetic. AB - Reports of waterskiing-related injuries are limited in the medical literature. The authors report a case of a unique waterskiing injury that progressed into a necrotizing foot infection in a well-controlled insulin-dependent diabetic. This case shows that with aggressive therapy, including hyperbaric oxygen treatments and use of a wound vacuum-assisted closure system, limbs can be saved. PMID- 16698913 TI - A role for vacuum-assisted closure in lower limb trauma: a proposed algorithm. AB - The British Association of Plastic Surgeons and British Orthopaedic Association estimate that 70% of Gustilo 3b tibial fractures require flap coverage. Prior to the introduction of vacuum-assisted closure (V.A.C.) in the late 1990s, options were limited for the treatment of such injuries in selected individuals with medical comorbidity or complex wounds. VAC has now become an important part of our algorithm for soft tissue reconstruction with excellent results, but its role has not yet been defined in common management pathways. The authors present 3 cases, all different in their mechanism of injury, type of injury, and subsequent management, but all had a successful outcome with VAC. PMID- 16698914 TI - Management of a difficult-to-heal chronic wound infected with methycillin resistant staphylococcus aureus in a patient with psoriasis following a complex knee surgery. AB - This report presents the difficulties encountered in managing a wound colonized with methycillin-resistant Staphylococcus aureus (MRSA) following a complex knee surgery in an elderly female patient with generalized psoriasis. The patient's chronic wound was successfully treated with nanocrystalline silver-releasing dressings (Acticoat). The patient did not develop a deep-seated infection, nor was removal of the implant needed. However, the wound infection persisted for 6 months. At 3 years follow-up, the patient has satisfactory skin over the implant on the knee. PMID- 16698915 TI - Evaluation of the antibacterial activity and toxicity of 2 new hydrogels: a pilot study. AB - Wound bed preparation remains a very important issue in wound healing. To promote the production of granulation tissue, it is necessary to remove necrotic tissue and to control infection. Necrotic tissue may be removed using a hydrogel preparation. Flaminal and Flaminal Hydro (Flen Pharma, Belgium) are 2 new hydroactive colloid gel dressings with state antibacterial properties. These properties are attributed to an enzymatic complex in their formulation. In the study described in this report, the antibacterial effects of Flaminal and Flaminal Hydro were confirmed in an in vitro as well as an in vivo setting. It was also demonstrated that Flaminal and Flaminal Hydro are not toxic to keratinocytes in vitro using an MTT [3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide] test. PMID- 16698917 TI - A case of cutaneous vasculitis associated with olanzapine. PMID- 16698918 TI - Local myocardial insulin-like growth factor 1 (IGF-1) delivery with biotinylated peptide nanofibers improves cell therapy for myocardial infarction. AB - Strategies for cardiac repair include injection of cells, but these approaches have been hampered by poor cell engraftment, survival, and differentiation. To address these shortcomings for the purpose of improving cardiac function after injury, we designed self-assembling peptide nanofibers for prolonged delivery of insulin-like growth factor 1 (IGF-1), a cardiomyocyte growth and differentiation factor, to the myocardium, using a "biotin sandwich" approach. Biotinylated IGF-1 was complexed with tetravalent streptavidin and then bound to biotinylated self assembling peptides. This biotin sandwich strategy allowed binding of IGF-1 but did not prevent self-assembly of the peptides into nanofibers within the myocardium. IGF-1 that was bound to peptide nanofibers activated Akt, decreased activation of caspase-3, and increased expression of cardiac troponin I in cardiomyocytes. After injection into rat myocardium, biotinylated nanofibers provided sustained IGF-1 delivery for 28 days, and targeted delivery of IGF-1 in vivo increased activation of Akt in the myocardium. When combined with transplanted cardiomyocytes, IGF-1 delivery by biotinylated nanofibers decreased caspase-3 cleavage by 28% and increased the myocyte cross-sectional area by 25% compared with cells embedded within nanofibers alone or with untethered IGF-1. Finally, cell therapy with IGF-1 delivery by biotinylated nanofibers improved systolic function after experimental myocardial infarction, demonstrating how engineering the local cellular microenvironment can improve cell therapy. PMID- 16698919 TI - On the hand-over-hand mechanism of kinesin. AB - We present here a simple theoretical model for conventional kinesin. The model reproduces the hand-over-hand mechanism for kinesin walking to the plus end of a microtubule. A large hindering force induces kinesin to walk slowly to the minus end, again by a hand-over-hand mechanism. Good agreement is obtained between the calculated and experimental results on the external force dependence of the walking speed, the forward/backward step ratio, and dwell times for both forward and backward steps. The model predicts that both forward and backward motions of kinesin take place at the same chemical state of the motor heads, with the front head being occupied by an ATP (or ADP,Pi) and the rear being occupied by an ADP. The direction of motion is a result of the competition between the power stroke produced by the front head and the external load. The other predictions include the external force dependence of the chemomechanical coupling ratio (e.g., the stepping distance/ATP ratio) and the walking speed of kinesin at force ranges that have not been tested by experiments. The model predicts that the chemomechanical coupling remains tight in a large force range. However, when the external force is very large (e.g., approximately 18 pN), kinesin slides in an inchworm fashion, and the translocation of kinesin becomes loosely coupled to ATP turnovers. PMID- 16698920 TI - Differentiation of proteins based on characteristic patterns of association and denaturation in solutions of SDS. AB - This paper shows that proteins display an unexpectedly wide range of behaviors in buffers containing moderate (0.1-10 mM) concentrations of SDS (complete unfolding, formation of stable intermediate states, specific association with SDS, and various kinetic phenomena); capillary electrophoresis provides a convenient method of examining these behaviors. Examination of the dynamics of the response of proteins to SDS offers a way to differentiate and characterize proteins. Based on a survey of 18 different proteins, we demonstrate that proteins differ in the concentrations of SDS at which they denature, in the rates of unfolding in SDS, and in the profiles of the denaturation pathways. We also demonstrate that these differences can be exploited in the analysis of mixtures. PMID- 16698921 TI - Runaway domain swapping in amyloid-like fibrils of T7 endonuclease I. AB - Amyloid fibrils are associated with >20 fatal human disorders, including Alzheimer's, Parkinson's, and prion diseases. Knowledge of how soluble proteins assemble into amyloid fibrils remains elusive despite its potential usefulness for developing diagnostics and therapeutics. In at least some fibrils, runaway domain swapping has been proposed as a possible mechanism for fibril formation. In runaway domain swapping, each protein molecule swaps a domain into the complementary domain of the adjacent molecule along the fibril. Here we show that T7 endonuclease I, a naturally domain-swapped dimeric protein, can form amyloid like fibrils. Using protein engineering, we designed a double-cysteine mutant that forms amyloid-like fibrils in which molecules of T7 endonuclease I are linked by intermolecular disulfide bonds. Because the disulfide bonds are designed to form only at the domain-swapped dimer interface, the resulting covalently linked fibrils show that T7 endonuclease I forms fibrils by a runaway domain swap. In addition, we show that the disulfide mutant exists in two conformations, only one of which is able to form fibrils. We also find that domain-swapped dimers, if locked in a close-ended dimeric form, are unable to form fibrils. Our study provides strong evidence for runaway domain swapping in the formation of an amyloid-like fibril and, consequently, a molecular explanation for specificity and stability of fibrils. In addition, our results suggest that inhibition of fibril formation for domain-swapped proteins may be achieved by stabilizing domain-swapped dimers. PMID- 16698922 TI - Hsk1 kinase is required for induction of meiotic dsDNA breaks without involving checkpoint kinases in fission yeast. AB - Cdc7 kinase, conserved through evolution, is known to be essential for mitotic DNA replication. The role of Cdc7 in meiotic recombination was suggested in Saccharomyces cerevisiae, but its precise role has not been addressed. Here, we report that Hsk1, the Cdc7-related kinase in Schizosaccharomyces pombe, plays a crucial role during meiosis. In a hsk1 temperature-sensitive strain (hsk1-89), meiosis is arrested with one nucleus state before meiosis I in most of the cells and meiotic recombination frequency is reduced by one order of magnitude, whereas premeiotic DNA replication is delayed but is apparently completed. Strikingly, formation of meiotic dsDNA breaks (DSBs) are largely impaired in the mutant, and Hsk1 kinase activity is essential for these processes. Deletion of all three checkpoint kinases, namely Cds1, Chk1, and Mek1, does not restore DSB formation, meiosis, or Cdc2 activation, which is suppressed in hsk1-89, suggesting that these aberrations are not caused by known checkpoint pathways but that Hsk1 may regulate DSB formation and meiosis. Whereas transcriptional induction of some rec genes and horsetail movement are normal, chromatin remodeling at ade6-M26, a recombination hotspot, which is prerequisite for subsequent DSB formation at this locus, is not observed in hsk1-89. These results indicate unique and essential roles of Hsk1 kinase in the initiation of meiotic recombination and meiosis. PMID- 16698923 TI - Tomato aromatic amino acid decarboxylases participate in synthesis of the flavor volatiles 2-phenylethanol and 2-phenylacetaldehyde. AB - An important phenylalanine-derived volatile compound produced by plants is 2 phenylethanol. It is a major contributor to flavor in many foods, including fresh fruits, such as tomato, and an insect-attracting scent in roses and many other flowers. Despite the centrality of 2-phenylethanol to flavor and fragrance, the plant genes responsible for its synthesis have not been identified. Here, we describe a biosynthetic pathway for 2-phenylethanol and other phenylalanine derived volatiles in tomato fruits and a small family of decarboxylases (LeAADC1A, LeAADC1B, and LeAADC2) that can mediate that pathway's first step. These enzymes each catalyze conversion of phenylalanine to phenethylamine and tyrosine to tyramine. Although tyrosine is the preferred substrate in vitro, phenylalanine levels in tomato fruits far exceed those of tyrosine, indicating that phenylalanine is a physiological substrate. Consistent with this view, overexpression of either LeAADC1A or LeAADC2 in transgenic tomato plants resulted in fruits with up to 10-fold increased emissions of the products of the pathway, including 2-phenylacetaldehyde, 2-phenylethanol, and 1-nitro-2-phenylethane. Further, antisense reduction of LeAADC2 significantly reduced emissions of these volatiles. Besides establishing a biosynthetic route, these results show that it is possible to change phenylalanine-based flavor and aroma volatiles in plants by manipulating expression of a single gene. PMID- 16698924 TI - Expression of 5-lipoxygenase and leukotriene A4 hydrolase in human atherosclerotic lesions correlates with symptoms of plaque instability. AB - Leukotrienes (LT) are a group of proinflammatory lipid mediators that are implicated in the pathogenesis and progression of atherosclerosis. Here we report that mRNA levels for the three key proteins in LTB4 biosynthesis, namely 5 lipoxygenase (5-LO), 5-LO-activating protein (FLAP), and LTA4 hydrolase (LTA4H), are significantly increased in human atherosclerotic plaque (n = 72) as compared with healthy controls (n = 6). Neither LTC4 synthase nor any of the LT receptors exhibits significantly increased mRNA levels. Immunohistochemical staining revealed abundant expression of 5-LO, FLAP, and LTA4H protein, colocalizing in macrophages of intimal lesions. Human lesion tissue converts arachidonic acid into significant amounts of LTB4, and a selective, tight-binding LTA4H inhibitor can block this activity. Furthermore, expression of 5-LO and LTA4H, but not FLAP, is increased in patients with recent or ongoing symptoms of plaque instability, and medication with warfarin correlates with increased levels of FLAP mRNA. In contrast to human plaques, levels of 5-LO mRNA are not significantly increased in plaque tissues from two atherosclerosis-prone mouse strains, and mouse plaques exhibit segregated cellular expression of LTA4H and 5-LO as well as strong increases of CysLT1 and CysLT2 mRNA. These discrepancies indicate that phenotypic changes in the synthesis and action of LT in specific mouse models of atherosclerosis should be cautiously translated into human pathology. The abundant expression of LTA4H and correlation with plaque instability identify LTA4H as a potential target for pharmacological intervention in treatment of human atherosclerosis. PMID- 16698925 TI - A heme-degradation pathway in a blood-sucking insect. AB - Hematophagous insects are vectors of diseases that affect hundreds of millions of people worldwide. A common physiological event in the life of these insects is the hydrolysis of host hemoglobin in the digestive tract, leading to a massive release of heme, a known prooxidant molecule. Diverse organisms, from bacteria to plants, express the enzyme heme oxygenase, which catalyzes the oxidative degradation of heme to biliverdin (BV) IX, CO, and iron. Here, we show that the kissing bug Rhodnius prolixus, a vector of Chagas' disease, has a unique heme degradation pathway wherein heme is first modified by addition of two cysteinylglycine residues before cleavage of the porphyrin ring, followed by trimming of the dipeptides. Furthermore, in contrast to most known heme oxygenases, which generate BV IXalpha, in this insect, the end product of heme detoxification is a dicysteinyl-BV IXgamma. Based on these results, we propose a heme metabolizing pathway that includes the identified intermediates produced during modification and cleavage of the heme porphyrin ring. PMID- 16698926 TI - L1 retrotransposition in nondividing and primary human somatic cells. AB - Whether long interspersed element-1 (L1 or LINE-1) retrotransposition can occur in quiescent, nondividing, and/or terminally differentiated somatic cells has remained an unanswered fundamental question in human genetics. Here, we used a ubiquitously active phosphoglycerate kinase-1 promoter to drive the expression of a highly active human L1 element from an adenovirus-L1 hybrid vector. This vector system achieved retrotransposition in up to 91% of actively growing immortalized cells, and we demonstrated that L1 retrotransposition can be suppressed by the reverse transcriptase inhibitor 3'-azido-3'-deoxythymidine. This adenovirus vector enabled efficient delivery of the L1 element into differentiated primary human somatic cells and G1/S-arrested cells, resulting in retrotransposition in both cases; however, it was not detected in G0-arrested cells. Thus, these data indicate that L1 retrotransposition can occur in nondividing somatic cells. PMID- 16698927 TI - kappa-casein-deficient mice fail to lactate. AB - Acquisition of milk production capabilities by an ancestor of mammals is at the root of mammalian evolution. Milk casein micelles are a primary source of amino acids and calcium phosphate to neonates. To understand the role of kappa-casein in lactation, we have created and characterized a null mouse strain (Csnk-/-) lacking this gene. The mutant kappa-casein allele did not affect the expression of other milk proteins in Csnk-/- females. However, these females did not suckle their pups and failed to lactate because of destabilization of the micelles in the lumina of the mammary gland. Thus, kappa-casein is essential for lactation and, consequently, for the successful completion of the process of reproduction in mammals. In view of the extreme structural conservation of the casein locus, as well as the phenotype of Csnk-/- females, we propose that the organization of a functional kappa-casein gene would have been one of the critical events in the evolution of mammals. Further, kappa-casein variants are known to affect the industrial properties of milk in dairy animals. Given the expenses and the time scale of such experiments in livestock species, it is desirable to model the intended genetic modifications in mice first. The mouse strain that we have created would be a useful model to study the effect of kappa-casein variants on the properties of milk and/or milk products. PMID- 16698928 TI - Backsteps induced by nucleotide analogs suggest the front head of kinesin is gated by strain. AB - The two-headed kinesin motor harnesses the energy of ATP hydrolysis to take 8-nm steps, walking processively along a microtubule, alternately stepping with each of its catalytic heads in a hand-over-hand fashion. Two persistent challenges for models of kinesin motility are to explain how the two heads are coordinated ("gated") and when the translocation step occurs relative to other events in the mechanochemical reaction cycle. To investigate these questions, we used a precision optical trap to measure the single-molecule kinetics of kinesin in the presence of substrate analogs beryllium fluoride or adenylyl-imidodiphosphate. We found that normal stepping patterns were interspersed with long pauses induced by analog binding, and that these pauses were interrupted by short-lived backsteps. After a pause, processive stepping could only resume once the kinesin molecule took an obligatory, terminal backstep, exchanging the positions of its front and rear heads, presumably to allow release of the bound analog from the new front head. Preferential release from the front head implies that the kinetics of the two heads are differentially affected when both are bound to the microtubule, presumably by internal strain that is responsible for the gating. Furthermore, we found that ATP binding was required to reinitiate processive stepping after the terminal backstep. Together, our results support stepping models in which ATP binding triggers the mechanical step and the front head is gated by strain. PMID- 16698929 TI - Selective regulatory function of Socs3 in the formation of IL-17-secreting T cells. AB - Suppressor of cytokine signaling (Socs) 3 is a cytokine-inducible inhibitor with critical but selective cell-specific effects. We show that deficiency of Socs3 in T cells had minimal effects on differentiation of T cells to the T helper (Th) 1 or Th2 subsets; accordingly, Socs3 had no effect on IL-12-dependent signal transducer and activator of transcription (Stat) 4 phosphorylation or IL-4 dependent Stat6 phosphorylation. By contrast, Socs3 was found to be a major regulator of IL-23-mediated Stat3 phosphorylation and Th17 generation, and Stat3 directly binds to the IL-17A and IL-17F promoters. We conclude that Socs3 is an essential negative regulator of IL-23 signaling, inhibition of which constrains the generation of Th17 differentiation. PMID- 16698930 TI - Low-dose alcohol actions on alpha4beta3delta GABAA receptors are reversed by the behavioral alcohol antagonist Ro15-4513. AB - Although it is now more than two decades since it was first reported that the imidazobenzodiazepine Ro15-4513 reverses behavioral alcohol effects, the molecular target(s) of Ro15-4513 and the mechanism of alcohol antagonism remain elusive. Here, we show that Ro15-4513 blocks the alcohol enhancement on recombinant "extrasynaptic" alpha4/6beta3delta GABA(A) receptors at doses that do not reduce the GABA-induced Cl(-) current. At low ethanol concentrations (< or =30 mM), the Ro15-4513 antagonism is complete. However, at higher ethanol concentrations (> or =100 mM), there is a Ro15-4513-insensitive ethanol enhancement that is abolished in receptors containing a point mutation in the second transmembrane region of the beta3 subunit (beta3N265M). Therefore, alpha4/6beta3delta GABA receptors have two distinct alcohol modulation sites: (i) a low-dose ethanol site present in alpha4/6beta3delta receptors that is antagonized by the behavioral alcohol antagonist Ro15-4513 and (ii) a site activated at high (anesthetic) alcohol doses, defined by mutations in membrane spanning regions. Receptors composed of alpha4beta3N265Mdelta subunits that lack the high-dose alcohol site show a saturable ethanol dose-response curve with a half-maximal enhancement at 16 mM, close to the legal blood alcohol driving limit in most U.S. states (17.4 mM). Like in behavioral experiments, the alcohol antagonist effect of Ro15-4513 on recombinant alpha4beta3delta receptors is blocked by flumazenil and beta-carboline-ethyl ester (beta-CCE). Our findings suggest that ethanol/Ro15-4513-sensitive GABA(A) receptors are important mediators of behavioral alcohol effects. PMID- 16698931 TI - Role of TL1A and its receptor DR3 in two models of chronic murine ileitis. AB - TL1A is a TNF-like cytokine that binds to the death-domain receptor (DR)3 and provides costimulatory signals to activated lymphocytes. Through this interaction, TL1A induces secretion of IFN-gamma and may, therefore, participate in the development of T helper-1-type effector responses. In this study, we investigated whether interactions between TL1A and DR3 are involved in the pathogenesis of chronic murine ileitis. We demonstrate that alternative splicing of DR3 mRNA takes place during the activation of lymphocytes, which results in up regulation of the complete/transmembrane (tm) form of DR3. Using two immunogenetically distinct animal models of Crohn's disease, we demonstrate that induction of intestinal inflammation is associated with significant up-regulation of TL1A and tm DR3 in the inflamed mucosa. In addition, within isolated lamina propria mononuclear cells from mice with inflammation, TL1A is primarily expressed on CD11c(high) dendritic cells. We also report that TL1A acts preferentially on memory CD4(+)/CD45RB(lo) murine lymphocytes by significantly inducing their proliferation, whereas it does not affect the proliferation of the naive CD4(+)/CD45RB(hi) T helper cell subpopulation. Finally, we demonstrate that TL1A synergizes with both the cytokine-dependent IL-12/IL-18 pathway and with low dose stimulation of the T cell receptor to significantly induce the secretion of IFN-gamma via an IL-18-independent pathway. Our results raise the possibility that interaction(s) between TL1A expressed on antigen-presenting cells and tm DR3 on lymphocytes may be of particular importance for the pathogenesis of chronic inflammatory conditions that depend on IFN-gamma secretion, including inflammatory bowel disease. Blockade of the TL1A/DR3 pathway may, therefore, offer therapeutic opportunities in Crohn's disease. PMID- 16698932 TI - Frequency-specific and D2 receptor-mediated inhibition of glutamate release by retrograde endocannabinoid signaling. AB - The mechanisms underlying modulation of corticostriatal synaptic transmission by D2-like receptors (D2Rs) have been controversial. A recent study suggested that D2Rs inhibit glutamate release at this synapse, but only during high-frequency synaptic activation. Because the release of postsynaptic endocannabinoids (eCBs), which act as retrograde messengers to inhibit presynaptic glutamate release, can be triggered by D2R activation and intense synaptic activation, such a mechanism could mediate dopaminergic modulation of corticostriatal transmission. Here, we show that D2R activation reduces excitatory transmission onto striatal medium spiny neurons at a stimulation frequency of 20 Hz but not at 1 Hz. This form of inhibition requires CB1 receptor activation, as evidenced by the fact that it is blocked by AM251 [N-(piperidin-1-yl)-1-(2,4-dichlorophenyl)-5-(4-chlorophenyl)-4 methyl-1H-pyrazole-3-carboxamide], a CB1 antagonist, and is absent in CB1 knockout mice. It is also blocked by postsynaptic intracellular calcium chelation, by group I metabotropic glutamate receptor antagonism, and by inhibition of postsynaptic phospholipase C. These results demonstrate a previously unrecognized role for retrograde eCB signaling in reversible and frequency-specific inhibition of glutamate release by the activation of striatal D2Rs. PMID- 16698933 TI - Hierarchical structures induce long-range dynamical correlations in written texts. AB - Thoughts and ideas are multidimensional and often concurrent, yet they can be expressed surprisingly well sequentially by the translation into language. This reduction of dimensions occurs naturally but requires memory and necessitates the existence of correlations, e.g., in written text. However, correlations in word appearance decay quickly, while previous observations of long-range correlations using random walk approaches yield little insight on memory or on semantic context. Instead, we study combinations of words that a reader is exposed to within a "window of attention," spanning about 100 words. We define a vector space of such word combinations by looking at words that co-occur within the window of attention, and analyze its structure. Singular value decomposition of the co-occurrence matrix identifies a basis whose vectors correspond to specific topics, or "concepts" that are relevant to the text. As the reader follows a text, the "vector of attention" traces out a trajectory of directions in this "concept space." We find that memory of the direction is retained over long times, forming power-law correlations. The appearance of power laws hints at the existence of an underlying hierarchical network. Indeed, imposing a hierarchy similar to that defined by volumes, chapters, paragraphs, etc. succeeds in creating correlations in a surrogate random text that are identical to those of the original text. We conclude that hierarchical structures in text serve to create long-range correlations, and use the reader's memory in reenacting some of the multidimensionality of the thoughts being expressed. PMID- 16698934 TI - Tetrahedral structure or chains for liquid water. AB - It has been suggested, based on x-ray absorption spectroscopy (XAS) experiments on liquid water [Wernet, Ph., et al. (2004) Science 304, 995-999], that a condensed-phase water molecule's asymmetric electron density results in only two hydrogen bonds per water molecule on average. The larger implication of the XAS interpretation is that the conventional view of liquid water being a tetrahedrally coordinated random network is now replaced by a structural organization that instead strongly favors hydrogen-bonded water chains or large rings embedded in a weakly hydrogen-bonded disordered network. This work reports that the asymmetry of the hydrogen density exhibited in the XAS experiments agrees with reported x-ray scattering structure factors and intensities for Q > 6.5 A(-1). However, the assumption that the asymmetry in the hydrogen electron density does not fluctuate and is persistent in all local molecular liquid water environments is inconsistent with longer-ranged tetrahedral network signatures present in experimental x-ray scattering intensity and structure factor data for Q < 6.5 A(-1). PMID- 16698935 TI - Genetic effects of chronic habitat fragmentation in a wind-pollinated tree. AB - Habitat fragmentation poses a serious threat to plants through genetic changes associated with increased isolation and reduced population size. However, the longevity of trees, combined with effective seed or pollen dispersal, can enhance their resistance to these effects. The European beech (Fagus sylvatica) dominates forest over large regions of Europe. We demonstrate that habitat fragmentation in this species has led to genetic bottlenecks and the disruption of the species' breeding system, leading to significantly elevated levels of inbreeding, population divergence, and reduced genetic diversity within populations. These results show that, in contrast with the findings of previous studies, forest fragmentation has a negative genetic impact, even in this widespread, wind pollinated tree. The identification of significant effects of forest fragmentation in beech demonstrates that trees are not at reduced risk from environmental change. This should be accounted for in the management of remaining natural and seminatural forest throughout the world. PMID- 16698936 TI - The maximum number of torque-generating units in the flagellar motor of Escherichia coli is at least 11. AB - Torque is generated in the rotary motor at the base of the bacterial flagellum by ion translocating stator units anchored to the peptidoglycan cell wall. Stator units are composed of the proteins MotA and MotB in proton-driven motors, and they are composed of PomA and PomB in sodium-driven motors. Strains of Escherichia coli lacking functional stator proteins produce flagella that do not rotate, and induced expression of the missing proteins leads to restoration of motor rotation in discrete speed increments, a process known as "resurrection." Early work suggested a maximum of eight units. More recent indications that WT motors may contain more than eight units, based on recovery of disrupted motors, are inconclusive. Here we demonstrate conclusively that the maximum number of units in a motor is at least 11. Using back-focal-plane interferometry of 1-mum polystyrene beads attached to flagella, we observed at least 11 distinct speed increments during resurrection with three different combinations of stator proteins in E. coli. The average torques generated by a single unit and a fully induced motor were lower than previous estimates. Speed increments at high numbers of units are smaller than those at low numbers, indicating that not all units in a fully induced motor are equivalent. PMID- 16698937 TI - Signature whistle shape conveys identity information to bottlenose dolphins. AB - Bottlenose dolphins (Tursiops truncatus) develop individually distinctive signature whistles that they use to maintain group cohesion. Unlike the development of identification signals in most other species, signature whistle development is strongly influenced by vocal learning. This learning ability is maintained throughout life, and dolphins frequently copy each other's whistles in the wild. It has been hypothesized that signature whistles can be used as referential signals among conspecifics, because captive bottlenose dolphins can be trained to use novel, learned signals to label objects. For this labeling to occur, signature whistles would have to convey identity information independent of the caller's voice features. However, experimental proof for this hypothesis has been lacking. This study demonstrates that bottlenose dolphins extract identity information from signature whistles even after all voice features have been removed from the signal. Thus, dolphins are the only animals other than humans that have been shown to transmit identity information independent of the caller's voice or location. PMID- 16698938 TI - PKCepsilon increases endothelin converting enzyme activity and reduces amyloid plaque pathology in transgenic mice. AB - Deposition of plaques containing amyloid beta (Abeta) peptides is a neuropathological hallmark of Alzheimer's disease (AD). Here we demonstrate that neuronal overexpression of the epsilon isozyme of PKC decreases Abeta levels, plaque burden, and plaque-associated neuritic dystrophy and reactive astrocytosis in transgenic mice expressing familial AD-mutant forms of the human amyloid precursor protein (APP). Compared with APP singly transgenic mice, APP/PKCepsilon doubly transgenic mice had decreased Abeta levels but showed no evidence for altered cleavage of APP. Instead, PKCepsilon overexpression selectively increased the activity of endothelin-converting enzyme, which degrades Abeta. The activities of other Abeta-degrading enzymes, insulin degrading enzyme and neprilysin, were unchanged. These results indicate that increased neuronal PKCepsilon activity can promote Abeta clearance and reduce AD neuropathology through increased endothelin-converting enzyme activity. PMID- 16698939 TI - Synaptic tetraspan vesicle membrane proteins are conserved but not needed for synaptogenesis and neuronal function in Caenorhabditis elegans. AB - Tetraspan vesicle membrane proteins (TVPs) comprise a major portion of synaptic vesicle proteins, yet their contribution to the synaptic vesicle cycle is poorly understood. TVPs are grouped in three mammalian gene families: physins, gyrins, and secretory carrier-associated membrane proteins (SCAMPs). In Caenorhabditis elegans, only a single member of each of these families exists. These three nematode TVPs colocalize to the same vesicular compartment when expressed in mammalian cells, suggesting that they could serve overlapping functions. To examine their function, C. elegans null mutants were isolated for each gene, and a triple mutant was generated. Surprisingly, these animals develop normally and exhibit normal neuronal architecture and synaptic contacts. In addition, functions of the motor and sensory systems are normal as determined by pharmacological, chemotaxis, and thermotaxis assays. Finally, direct electrophysiological analysis of the neuromuscular junction revealed no phenotype in the TVP mutants. We therefore conclude that TVPs are not needed for the basic neuronal machinery and instead may contribute to subtle higher order functions. PMID- 16698940 TI - The immune response in autism: a new frontier for autism research. AB - Autism spectrum disorders (ASD) are part of a broad spectrum of neurodevelopmental disorders known as pervasive developmental disorders, which occur in childhood. They are characterized by impairments in social interaction, verbal and nonverbal communication and the presence of restricted and repetitive stereotyped behaviors. At the present time, the etiology of ASD is largely unknown, but genetic, environmental, immunological, and neurological factors are thought to play a role in the development of ASD. Recently, increasing research has focused on the connections between the immune system and the nervous system, including its possible role in the development of ASD. These neuroimmune interactions begin early during embryogenesis and persist throughout an individual's lifetime, with successful neurodevelopment contingent upon a normal balanced immune response. Immune aberrations consistent with a dysregulated immune response, which so far, have been reported in autistic children, include abnormal or skewed T helper cell type 1 (T(H)1)/T(H)2 cytokine profiles, decreased lymphocyte numbers, decreased T cell mitogen response, and the imbalance of serum immunoglobulin levels. In addition, autism has been linked with autoimmunity and an association with immune-based genes including human leukocyte antigen (HLA)-DRB1 and complement C4 alleles described. There is potential that such aberrant immune activity during vulnerable and critical periods of neurodevelopment could participate in the generation of neurological dysfunction characteristic of ASD. This review will examine the status of the research linking the immune response with ASD. PMID- 16698941 TI - New insights into the regulation of TLR signaling. AB - Toll-like receptor (TLR) activation is dictated by a number of factors including the ligand itself and the localization of the receptor, in terms of expression profile and subcellular localization and the signal transduction pathway that has been activated. Recent work into TLR signal transduction has revealed complex regulation at a number of different levels including regulation by phosphorylation, targeted degradation, and sequestration of signaling molecules. Here, we describe recent advances that have been made in our understanding of how TLR signaling is regulated at the biochemical level. PMID- 16698942 TI - Cytokines, chemokines, and their receptors: targets for immunomodulation. Conference report: International Cytokine Society Conference 2005. AB - The International Cytokine Society Conference 2005 was held in Seoul, Korea, October 27-31, 2005. This meeting provided the attendants an excellent forum to discuss recent advances in the biology of cytokines, chemokines, their receptors, and the receptors for innate immunity, with particular emphasis on the possibility of interfering with these systems for therapeutic purposes. We herein summarize some of the key findings discussed at the meeting. PMID- 16698943 TI - Comparative gene expression analysis of NKT cell subpopulations. AB - Natural killer T (NKT) cells are a lymphocyte lineage, which has diverse immune regulatory activities in many disease settings. Most previous studies have investigated the functions of this family of cells as a single entity, but more recent evidence highlights the distinct functional and phenotypic properties of NKT cell subpopulations. It is likely that the diverse functions of NKT cells are regulated and coordinated by these different NKT subsets. Little is known about how NKT subsets differ in their interactions with the host. We have undertaken the first microarray analysis comparing the gene expression profiles of activated human NKT cell subpopulations, including CD8(+) NKT cells, which have often been overlooked. We describe the significant gene expression differences among NKT cell subpopulations and some of the molecules likely to confer their distinct functional roles. Several genes not associated previously with NKT cells were shown to be expressed differentially in specific NKT cell subpopulations. Our findings provide new insights into the NKT cell family, which may direct further research toward better manipulation of NKT cells for therapeutic applications. PMID- 16698944 TI - Arabidopsis PASTICCINO2 is an antiphosphatase involved in regulation of cyclin dependent kinase A. AB - PASTICCINO2 (PAS2), a member of the protein Tyr phosphatase-like family, is conserved among all eukaryotes and is characterized by a mutated catalytic site. The cellular functions of the Tyr phosphatase-like proteins are still unknown, even if they are essential in yeast and mammals. Here, we demonstrate that PAS2 interacts with a cyclin-dependent kinase (CDK) that is phosphorylated on Tyr and not with its unphosphorylated isoform. Phosphorylation of the conserved regulatory Tyr-15 is involved in the binding of CDK to PAS2. Loss of the PAS2 function dephosphorylated Arabidopsis thaliana CDKA;1 and upregulated its kinase activity. In accordance with its role as a negative regulator of the cell cycle, overexpression of PAS2 slowed down cell division in suspension cell cultures at the G2-to-M transition and early mitosis and inhibited Arabidopsis seedling growth. The latter was accompanied by altered leaf development and accelerated cotyledon senescence. PAS2 was localized in the cytoplasm of dividing cells but moved into the nucleus upon cell differentiation, suggesting that the balance between cell division and differentiation is regulated through the interaction between CDKA;1 and the antiphosphatase PAS2. PMID- 16698945 TI - Gamma-tubulin is essential for microtubule organization and development in Arabidopsis. AB - The process of microtubule nucleation in plant cells is still a major question in plant cell biology. gamma-Tubulin is known as one of the key molecular players for microtubule nucleation in animal and fungal cells. Here, we provide genetic evidence that in Arabidopsis thaliana, gamma-tubulin is required for the formation of spindle, phragmoplast, and cortical microtubule arrays. We used a reverse genetics approach to investigate the role of the two Arabidopsis gamma tubulin genes in plant development and in the formation of microtubule arrays. Isolation of mutants in each gene and analysis of two combinations of gamma tubulin double mutants showed that the two genes have redundant functions. The first combination is lethal at the gametophytic stage. Disruption of both gamma tubulin genes causes aberrant spindle and phragmoplast structures and alters nuclear division in gametophytes. The second combination of gamma-tubulin alleles affects late seedling development, ultimately leading to lethality 3 weeks after germination. This partially viable mutant combination enabled us to follow dynamically the effects of gamma-tubulin depletion on microtubule arrays in dividing cells using a green fluorescent protein marker. These results establish the central role of gamma-tubulin in the formation and organization of microtubule arrays in Arabidopsis. PMID- 16698946 TI - SCARFACE encodes an ARF-GAP that is required for normal auxin efflux and vein patterning in Arabidopsis. AB - To identify molecular mechanisms controlling vein patterns, we analyzed scarface (sfc) mutants. sfc cotyledon and leaf veins are largely fragmented, unlike the interconnected networks in wild-type plants. SFC encodes an ADP ribosylation factor GTPase activating protein (ARF-GAP), a class with well-established roles in vesicle trafficking regulation. Quadruple mutants of SCF and three homologs (ARF-GAP DOMAIN1, 2, and 4) showed a modestly enhanced vascular phenotype. Genetic interactions between sfc and pinoid and between sfc and gnom suggest a possible function for SFC in trafficking of auxin efflux regulators. Genetic analyses also revealed interaction with cotyledon vascular pattern2, suggesting that lipid-based signals may underlie some SFC ARF-GAP functions. To assess possible roles for SFC in auxin transport, we analyzed sfc roots, which showed exaggerated responses to exogenous auxin and higher auxin transport capacity. To determine whether PIN1 intracellular trafficking was affected, we analyzed PIN1:green fluorescent protein (GFP) dynamics using confocal microscopy in sfc roots. We found normal PIN1:GFP localization at the apical membrane of root cells, but treatment with brefeldin A resulted in PIN1 accumulating in smaller and more numerous compartments than in the wild type. These data suggest that SFC is required for normal intracellular transport of PIN1 from the plasma membrane to the endosome. PMID- 16698947 TI - Spatial organisation of microbiota in quiescent adenoiditis and tonsillitis. AB - BACKGROUND: The reasons for recurrent adenotonsillitis are poorly understood. METHODS: The in situ composition of microbiota of nasal (5 children, 25 adults) and of hypertrophied adenoid and tonsillar tissue (50 children, 20 adults) was investigated using a broad range of fluorescent oligonucleotide probes targeted to bacterial rRNA. None of the patients had clinical signs of infection at the time of surgery. RESULTS: Multiple foci of ongoing purulent infections were found within hypertrophied adenoid and tonsillar tissue in 83% of patients, including islands and lawns of bacteria adherent to the epithelium, with concomitant marked inflammatory response, fissures filled with bacteria and pus, and diffuse infiltration of the tonsils by bacteria, microabscesses, and macrophages containing phagocytosed microorganisms. Haemophilusinfluenzae mainly diffusely infiltrated the tissue, Streptococcus and Bacteroides were typically found in fissures, and Fusobacteria,Pseudomonas and Burkholderia were exclusively located within adherent bacterial layers and infiltrates. The microbiota were always polymicrobial. CONCLUSIONS: Purulent processes persist during asymptomatic periods of adenotonsillitis. Most bacteria involved in this process are covered by a thick inflammatory infiltrate, are deeply invading, or are located within macrophages. The distribution of the bacteria within tonsils may be responsible for the failure of antibiotic treatment. PMID- 16698948 TI - The role of immunohistochemistry for smooth-muscle actin, p63, CD10 and cytokeratin 14 in the differential diagnosis of papillary lesions of the breast. AB - BACKGROUND: Histological differentiation of mammary papillary lesions can be difficult. The evaluation of myoepithelial cells can be helpful, with benign papilloma showing a continuous myoepithelial cell layer, which becomes attenuated or absent in malignant papillary lesions. METHODS: A large series of 100 papillomas (28 papillomas with florid epithelial hyperplasia) and 68 papillary carcinomas (9 invasive, 44 in situ, and 15 ductal carcinomas in situ (DCIS) involving papillomas) of the breast were stained for myoepithelial cells by immunohistochemistry using antibodies to smooth-muscle actin (SMA), p63, CD10 and cytokeratin (CK) 14. RESULTS: In the papillomas, using these four antibodies, myoepithelial cells were positive in 88%, 99%, 91% and 95% of cases, respectively, with SMA showing marked stromal component cell staining and CD10 showing epithelial and stromal staining. CK14 also showed epithelial staining in 71% of papillomas and 96% of papillomas with florid epithelial hyperplasia. In the papillary carcinomas, 36 (53%) cases showed staining of myoepithelial cells that were scattered, discontinuous and diminished in number and the remaining 32 (47%) cases did not show myoepithelial cells. Invasive papillary carcinoma has the lowest proportion (33%) with myoepithelial cells, and DCIS involving papillomas had the highest proportion (87%). CONCLUSIONS: p63 had the highest sensitivity and did not cross-react with stromal cells and only rarely with epithelial cells. CK14 has the added ability to distinguish between florid epithelial hyperplasia involving papilloma and DCIS involving papillomas. CK14 and p63 may be used as an adjunct in assessing difficult papillary lesions of the breast. PMID- 16698949 TI - Expression of KAI1 and tenascin, and microvessel density are closely correlated with liver metastasis of gastrointestinal adenocarcinoma. AB - AIM: To seek good markers to predict invasion and metastasis of gastrointestinal adenocarcinoma (GIA). METHODS: Expression of KAI1 and tenascin were examined on tissue microarrays containing gastric adenocarcinoma (n = 98), colorectal adenocarcinoma (n = 125), gastric adjacent non-cancerous mucosa (n = 95) and colorectal adjacent non-cancerous mucosa (n = 112) by immunostaining. Microvessel density (MVD) in GIA was labelled using anti-CD34 antibody by immunostaining. Expression of KAI1 and tenascin, and MVD were compared with clinicopathological features of tumours, including PTEN (phosphatase and tensin homology deleted from human chromosome 10) and EMMPRIN (extracellular matrix metalloproteinase inducer) expression. RESULTS: KAI1 expression was higher in GIAs than in their adjacent non-cancerous mucosa (p<0.05). KAI1 and tenascin expression showed a significantly negative association with liver metastasis of GIA (p<0.05), but not with depth of invasion, venous invasion or lymph node metastasis (p>0.05). A significantly negative relationship was observed between EMMPRIN and tenascin expression in GIA (p<0.05). MVD was positively correlated with depth of invasion, venous invasion, lymph node metastasis and liver metastasis of tumours (p<0.05), whereas it was negatively correlated with PTEN expression (p<0.05). CONCLUSIONS: Up-regulated KAI1 expression may play an important part in malignant transformation of gastrointestinal epithelial cells. Reduced expression of KAI1 and tenascin might underlie the molecular basis of liver metastasis of GIA. Angiogenesis is a key event in the invasion and metastasis of GIA. These markers might be used to indicate liver metastasis of GIA in clinicopathological practice. PMID- 16698951 TI - Variability of grade and stage in simultaneous paired liver biopsies in patients with hepatitis C. AB - BACKGROUND: Grading and staging of liver biopsies in patients with chronic hepatitis remains an inexact "gold standard" that is influenced by variabilities in scoring systems, sampling, observer agreement and expertise. Spatial disease variability relative to markers of the adequacy of biopsy has not been studied previously. METHODS: Paired liver biopsy specimens were obtained from the right and left hepatic lobes of 60 patients with chronic hepatitis C. Histological grade and disease stage were assessed according to the Ludwig scoring system, and scores were evaluated in relation to differences in size and number of portal tracts in all paired samples. RESULTS: The relative difference (%) in aggregate biopsy size and number of portal tracts was similar between paired samples with and without a difference in grade. Paired samples with a difference in stage showed a larger relative difference in biopsy size (p = 0.09) and in the number of portal tracts (p = 0.016). CONCLUSIONS: Our study shows a difference of one grade or one stage in 30% of paired liver biopsies, due to a combination of sampling variability and observer variability. Acknowledgment of "built-in" variability in grading and staging chronic hepatitis C by both clinicians and pathologists is essential for managing the individual patient with chronic hepatitis C. PMID- 16698950 TI - Value of ezrin, maspin and nm23-H1 protein expressions in predicting outcome of patients with head and neck squamous-cell carcinoma treated with radical radiotherapy. AB - BACKGROUND: Prognostic factors in predicting outcomes in patients with head and neck squamous-cell carcinoma (HNSCC) are limited to the clinical-pathological parameters, including lymph node metastasis, location, grade and stage of the disease. AIM: To determine whether the expression of these proteins has a value in predicting patient outcome. METHODS: Ezrin, maspin and nm23-H1 immunohistochemistry in tissue samples of 120 patients with HNSCC were evaluated using the microarray technique. RESULTS: In determining the association among each of the three proteins and the clinical-pathological parameters, low maspin expression was the only one found to be significantly associated with high tumour grade (p = 0.007); all others showed no significant associations. In univariate analysis, patients with tumours expressing high ezrin had a shorter disease-free survival (DFS) of 51% than those with low ezrin expression (DFS 84%; p = 0.08). In multivariate analysis, tumours with the combination of loss of maspin and low histological grade had longer DFS (83%) compared with those with high maspin and high histological grade (DFS 42%; p = 0.08). CONCLUSION: Our study is the first to determine the value of ezrin and maspin in HNSCC in a large series of patients with long follow-up. Ezrin and maspin seem to have a potential prognostic value in patients with HNSCC but results should be confirmed with further studies. PMID- 16698952 TI - Presence of mouse mammary tumour-like virus gene sequences may be associated with morphology of specific human breast cancer. AB - BACKGROUND: Mouse mammary tumour virus (MMTV) has a proven role in breast carcinogenesis in wild mice and genetically susceptible in-bred mice. MMTV-like env gene sequences, which indicate the presence of a replication-competent MMTV like virus, have been identified in some human breast cancers, but rarely in normal breast tissues. However, no evidence for a causal role of an MMTV-like virus in human breast cancer has emerged, although there are precedents for associations between specific histological characteristics of human cancers and the presence of oncogenic viruses. AIM: To investigate the possibility of an association between breast cancer and MMTV-like viruses. METHODS: Histological characteristics of invasive ductal human breast cancer specimens were compared with archival MMTV-associated mammary tumours from C3H experimental mice. The presence of MMTV-like env DNA sequences in the human breast cancer specimens was determined by polymerase chain reaction and confirmed by Southern hybridisation. RESULTS: MMTV-like env gene sequences were identified in 22 of 59 (37.3%) human breast cancer specimens. Seventeen of 43 (39.5%) invasive ductal carcinoma breast cancer specimens and 4 of 16 (25%) ductal carcinoma in situ specimens had some histological characteristics, which were similar to MMTV-associated mouse mammary tumours. However, these similarities were not associated with the presence or absence of MMTV-like gene sequences in the human breast tumour specimens. A significant (p = 0.05) correlation was found between the grade of the human breast cancer and similarity to the mouse mammary tumours. The lower the grade, the greater the similarity. CONCLUSION: Some human breast cancer specimens, in which MMTV-like env DNA sequences have been identified, were shown to have histological characteristics (morphology) similar to MMTV-associated mouse mammary tumours. These observations are compatible with, but not conclusive of, an association between the presence of MMTV-like env DNA sequences and some human breast cancers. PMID- 16698953 TI - Construction and validation of a bone marrow tissue microarray. AB - BACKGROUND: The use of tissue microarrays (TMAs) is now a generally accepted method for the investigation of solid tumours. However, little is known about the applicability of the TMA technique for analysis of patients with acute leukaemia. A bone marrow (BM)-TMA analysis with 15 different immunohistochemical markers was performed. The TMA was validated by comparison with the corresponding full tissue sections. MATERIALS AND METHODS: A BM-TMA comprising 148 cases of acute leukaemia, including 115 acute myeloid leukaemia (AML) and 33 acute lymphoblastic leukaemia (ALL) cases, was constructed. Expression of CD3, CD10, CD15, CD20, CD34, CD61, CD68, CD79a, CD99, CD117, CD138, myeloperoxidase, haemoglobin A1, glycophorin and terminal deoxynucleotidyl transferase was immunohistochemically analysed. 50 cases of the TMA were directly compared with the corresponding full tissue section to validate the results. RESULTS: Morphologically and immunohistochemically, 6 (4%) of 148 cases and 765 (11%) cores of 6912 individual analyses were not evaluable. A direct comparison of TMA cases with conventional full sections showed a concordance of the results of 100%. CONCLUSIONS: The small size of bone-marrow biopsies and the presence of bony trabeculae do not preclude construction and analysis of acute leukaemia TMAs. Acute leukaemia cases on TMA displayed the characteristic phenotypic profiles expected in different AML and ALL subtypes. Therefore, the TMA technique is also a promising method for high throughput analysis of combined marker expression and clinicopathological correlations in patients with leukaemia. PMID- 16698954 TI - Expression of peroxisome proliferator activated receptor gamma and cyclo oxygenase 2 in primary and recurrent ovarian carcinoma. AB - AIM: Peroxisome proliferator-activated receptor gamma (PPARgamma) has emerged as a potential therapeutic target in several types of cancer. In ovarian carcinomas, limited and conflicting data on PPARgamma protein expression have been reported. METHODS: The immunoexpression of PPARgamma and its putative target cyclo oxygenase 2 (COX2) was investigated in tumour tissues from 80 patients with primary and corresponding recurrent ovarian serous carcinomas after conventional platinum-based chemotherapy. RESULTS: PPARgamma expression was observed in 29% of primary and recurrent carcinomas. In the recurrent tumours, PPARgamma expression inversely correlated with COX2 overexpression in both chemosensitive (p = 0.02) and chemoresistant (p = 0.04) carcinomas. CONCLUSIONS: The data indicate that PPARgamma may represent a potential target for second-line treatment in ovarian cancers. PMID- 16698955 TI - Examination of peripheral blood films using automated microscopy; evaluation of Diffmaster Octavia and Cellavision DM96. AB - BACKGROUND: Differential counting of peripheral blood cells is an important diagnostic tool. Yet, this technique requires highly trained staff, is labour intensive and has limited statistical reliability. A recent development in this field was the introduction of automated peripheral blood differential counting systems. These computerised systems provide an automated morphological analysis of peripheral blood films, including a preclassification of both red and white cells (RBCs and WBCs, respectively). AIMS: To investigate the ability of two automated microscopy systems to examine peripheral blood smears. METHODS: Two automated microscopy systems, the Cellavision Diffmaster Octavia (Octavia) and Cellavision DM96 (DM96), were evaluated. RESULTS: The overall preclassification accuracy values for the Octavia and the DM96 systems were 87% and 92%, respectively. Evaluation of accuracy (WBC analysis) showed good correlation for both automated systems when compared with manual differentiation. Total analysis time (including post classification) was 5.4 min/slide for the Octavia and 3.2 min/slide for the DM96 (100 WBC/slide) system. The DM96 required even less time than manual differentiation by an experienced biomedical scientist. CONCLUSIONS: The Octavia and the DM96 are automated cell analysis systems capable of morphological classification of RBCs and WBCs in peripheral blood smears. Classification accuracy depends on the type of pathological changes in the blood sample. Both systems operate most effectively in the analysis of non-pathological blood samples. PMID- 16698956 TI - My approach to pancreatic fine needle aspiration. AB - Pancreatic fine needle aspiration cytopathology has earned a reputation as a rapid, safe, accurate and cost-beneficial modality of investigation of pancreatic mass lesion. Optimal results can be expected only if these procedures are undertaken regularly in gastroenterology departments processing large numbers of patients; occasional aspirators cannot exploit the technique to full potential. Professional teams following a dedicated approach to patient selection and management develop requisite expertise over time. Cytodiagnosis rests as much on morphological examination as on the milieu in which is it practised, and as much on sample quality as on cellular criteria. This article focuses attention on specimen handling, with particular reference to rapid on-site evaluation. The significance of particular cytodiagnoses in patient care is evaluated, and tumour types that may be encountered are enumerated and illustrated. PMID- 16698957 TI - Nuclear and cytoplasmic Maspin expression in primary non-small cell lung cancer. AB - AIM: To investigate whether nuclear and cytoplasmic Maspin expression is associated with distinct clinicopathological parameters and TP53 expression in a representative series of primary non-small cell lung cancer (NSCLC). METHODS: Tissue microarrays (n=487) were used to immunohistochemically analyse expression of Maspin and TP53. Cytoplasmic and nuclear expression of Maspin was scored on the basis of the percentage of positive tumour cells. Univariate analysis of clinicopathological variables potentially affecting tumour-specific survival was performed. RESULTS: Immunohistochemical Maspin expression (nuclear and cytoplasmic) was informative in 72.3% (352/487) of cases. Cytoplasmic and nuclear Maspin immunoreactivity in >or=10% of tumour cells was detected in 37.8% (133/352) and 65.3% (230/352) of informative cases, respectively. Nuclear and cytoplasmic Maspin staining was observed more frequently in primary squamous cell carcinomas than in other lung cancer types. Only nuclear Maspin immunoreactivity was significantly associated with positive TP53 staining. Cytoplasmic or nuclear Maspin expression was not associated with tumour-specific survival. CONCLUSION: Maspin expression was found both in the nucleus and the cytoplasm of NSCLC, more frequently in squamous cell carcinomas. However, no association with tumour specific survival could be demonstrated. PMID- 16698958 TI - MicroRNA enrichment among short 'ultraconserved' sequences in insects. AB - MicroRNAs are short (approximately 22 nt) regulatory RNA molecules that play key roles in metazoan development and have been implicated in human disease. First discovered in Caenorhabditis elegans, over 2500 microRNAs have been isolated in metazoans and plants; it has been estimated that there may be more than a thousand microRNA genes in the human genome alone. Motivated by the experimental observation of strong conservation of the microRNA let-7 among nearly all metazoans, we developed a novel methodology to characterize the class of such strongly conserved sequences: we identified a non-redundant set of all sequences 20 to 29 bases in length that are shared among three insects: fly, bee and mosquito. Among the few hundred sequences greater than 20 bases in length are close to 40% of the 78 confirmed fly microRNAs, along with other non-coding RNAs and coding sequence. PMID- 16698959 TI - GREM, a technique for genome-wide isolation and quantitative analysis of promoter active repeats. AB - We developed a technique called GREM (Genomic Repeat Expression Monitor) that can be applied to genome-wide isolation and quantitative analysis of any kind of transcriptionally active repetitive elements. Briefly, the technique includes three major stages: (i) generation of a transcriptome wide library of cDNA 5' terminal fragments, (ii) selective amplification of repeat-flanking genomic loci and (iii) hybridization of the cDNA library (i) to the amplicon (ii) with subsequent selective amplification and cloning of the cDNA-genome hybrids. The sequences obtained serve as 'tags' for promoter active repetitive elements. The advantage of GREM is an unambiguous mapping of individual promoter active repeats at a genome-wide level. We applied GREM for genome-wide experimental identification of human-specific endogenous retroviruses and their solitary long terminal repeats (LTRs) acting in vivo as promoters. Importantly, GREM tag frequencies linearly correlated with the corresponding LTR-driven transcript levels found using RT-PCR. The GREM technique enabled us to identify 54 new functional human promoters created by retroviral LTRs. PMID- 16698960 TI - Single-cell chromosomal imbalances detection by array CGH. AB - Genomic imbalances are a major cause of constitutional and acquired disorders. Therefore, aneuploidy screening has become the cornerstone of preimplantation, prenatal and postnatal genetic diagnosis, as well as a routine aspect of the diagnostic workup of many acquired disorders. Recently, array comparative genomic hybridization (array CGH) has been introduced as a rapid and high-resolution method for the detection of both benign and disease-causing genomic copy-number variations. Until now, array CGH has been performed using a significant quantity of DNA derived from a pool of cells. Here, we present an array CGH method that accurately detects chromosomal imbalances from a single lymphoblast, fibroblast and blastomere within a single day. Trisomy 13, 18, 21 and monosomy X, as well as normal ploidy levels of all other chromosomes, were accurately determined from single fibroblasts. Moreover, we showed that a segmental deletion as small as 34 Mb could be detected. Finally, we demonstrated the possibility to detect aneuploidies in single blastomeres derived from preimplantation embryos. This technique offers new possibilities for genetic analysis of single cells in general and opens the route towards aneuploidy screening and detection of unbalanced translocations in preimplantation embryos in particular. PMID- 16698961 TI - Strong physical constraints on sequence-specific target location by proteins on DNA molecules. AB - Sequence-specific binding to DNA in the presence of competing non-sequence specific ligands is a problem faced by proteins in all organisms. It is akin to the problem of parking a truck at a loading bay by the side of a road in the presence of cars parked at random along the road. Cars even partially covering the loading bay prevent correct parking of the truck. Similarly on DNA, non specific ligands interfere with the binding and function of sequence-specific proteins. We derive a formula for the probability that the loading bay is free from parked cars. The probability depends on the size of the loading bay and allows an estimation of the size of the footprint on the DNA of the sequence specific protein by assaying protein binding or function in the presence of increasing concentrations of non-specific ligand. Assaying for function gives an 'activity footprint'; the minimum length of DNA required for function rather than the more commonly measured physical footprint. Assaying the complex type I restriction enzyme, EcoKI, gives an activity footprint of approximately 66 bp for ATP hydrolysis and 300 bp for the DNA cleavage function which is intimately linked with translocation of DNA by EcoKI. Furthermore, considering the coverage of chromosomal DNA by proteins in vivo, our theory shows that the search for a specific DNA sequence is very difficult; most sites are obscured by parked cars. This effectively rules out any significant role in target location for mechanisms invoking one-dimensional, linear diffusion along DNA. PMID- 16698962 TI - Cloning and expression of new microRNAs from zebrafish. AB - MicroRNAs (miRNAs) play an important role in development and regulate the expression of many animal genes by post-transcriptional gene silencing. Here we describe the cloning and expression of new miRNAs from zebrafish. By high throughput sequencing of small-RNA cDNA libraries from 5-day-old zebrafish larvae and adult zebrafish brain we found 139 known miRNAs and 66 new miRNAs. For 65 known miRNAs and for 11 new miRNAs we also cloned the miRNA star sequence. We analyzed the temporal and spatial expression patterns for 35 new miRNAs and for 32 known miRNAs in the zebrafish by whole mount in situ hybridization and northern blotting. Overall, 23 of the 35 new miRNAs and 30 of the 32 known miRNAs could be detected. We found that most miRNAs were expressed during later stages of development. Some were expressed ubiquitously, but many of the miRNAs were expressed in a tissue-specific manner. Most newly discovered miRNAs have low expression levels and are less conserved in other vertebrate species. Our cloning and expression analysis indicates that most abundant and conserved miRNAs in zebrafish are now known. PMID- 16698964 TI - Estimated fluence-to-absorbed dose conversion coefficients for use in radiological protection of embryo and foetus against external exposure to photons from 50 keV to 10 GeV. AB - In the literature, no conversion coefficients are available for use in radiological protection of the embryo and foetus against external exposure to photons. This study used the Monte-Carlo code MCNPX to determine mean absorbed doses to the embryo and foetus when the mother is exposed to external photon fields. Monoenergetic photons ranging from 50 keV to 10 GeV were considered. The irradiation geometries included antero-posterior (AP), postero-anterior (PA), lateral (LAT), rotational (ROT), and isotropic (ISO). At each of these standard irradiation geometries, absorbed doses to the foetal brain and body were calculated for the embryo of 8 weeks and the foetus of 3, 6 or 9 months. Photon fluence-to-absorbed-dose conversion coefficients were estimated for the four prenatal ages. PMID- 16698963 TI - GAS41 interacts with transcription factor AP-2beta and stimulates AP-2beta mediated transactivation. AB - Transcription factor AP-2 regulates transcription of a number of genes involving mammalian development, differentiation and carcinogenesis. Recent studies have shown that interaction partners can modulate the transcriptional activity of AP-2 over the downstream targets. In this study, we reported the identification of GAS41 as an interaction partner of AP-2beta. We documented the interaction both in vivo by co-immunoprecipitation as well as in vitro through glutathione S transferase (GST) pull-down assays. We also showed that the two proteins are co localized in the nuclei of mammalian cells. We further mapped the interaction domains between the two proteins to the C-termini of both AP-2beta and GAS41, respectively. Furthermore, we have identified three critical residues of GAS41 that are important for the interaction between the two proteins. In addition, by transient co-expression experiments using reporter containing three AP-2 consensus binding sites in the promoter region, we found that GAS41 stimulates the transcriptional activity of AP-2beta over the reporter. Finally, electrophoretic mobility shift assay (EMSA) suggested that GAS41 enhances the DNA binding activity of AP-2beta. Our data provide evidence for a novel cellular function of GAS41 as a transcriptional co-activator for AP-2beta. PMID- 16698965 TI - Radioactivity concentrations and dose assessment for soil samples from Kestanbol granite area, Turkey. AB - The concentrations and distribution of natural and anthropogenic radionuclides in soils from around the Kestanbol (Canakkale), Turkey were investigated with an aim of evaluating the environmental radioactivity and radiological health hazard. Concentrations of radionuclides in the samples were determined by gamma ray spectrometer using HPGe detector. In the soils in general, the concentration of (232)Th was found to be higher than that of the (238)U and the activities of (232)Th and (238)U in this area are higher than the world average. (137)Cs was observed in all the samples, ranging from 0.37 +/- 0.22 to 36.03 +/- 0.54 Bq kg( 1). The mean radium equivalent activity, external hazard index and terrestrial absorbed dose rate for the area under study are 498 Bq kg(-1), 1.4 and 219 nGy h( 1), respectively. The annual effective dose to the public was found to be 269 muSv. The present data were compared with data obtained from different countries. PMID- 16698966 TI - Long-term measurements of H*(10) at aviation altitudes in the northern hemisphere. AB - Monitoring the radiation field at aviation altitudes is achieved by the dosemeter system piDOS installed in a passenger aircraft. The basic detector is a 2 in. tissue-equivalent proportional counter (TEPC) mounted in a cabin-baggage-sized aluminium suitcase. The entire system was characterised in neutron and photon reference fields from which two calibration factors were determined according to the splitting of the measured dose deposition spectrum yd(y) into low-LET and high-LET regions. A total of 255 flights in the northern hemisphere was analysed. The dependencies of the ambient dose equivalent rates on altitude, latitude and solar activity have been determined. These new data extend the data base used in Germany for the validation of program codes to calculate the radiation exposure of air crew members. PMID- 16698967 TI - Determination of ambient and personal dose equivalent for personnel and cargo security screening. AB - In the past few years, imaging technology using ionising radiation has been gaining in importance for the screening of goods and persons for security reasons and in order to detect contraband. For radiation protection purposes it is extremely important to know that dose persons are exposed to when passing through a personnel scanner or, as a stowaway, in a cargo scanner, so as to remain within the prescribed dose limits. Within the scope of a research project, measurements were performed on different types of personnel X-ray scanners as well as cargo X ray scanners, using the transmission and/or the backscattering method. All scanners investigated operate with a high dose rate and use short irradiation time. Owing to this method of scanning reliable values can only be determined for the personal and ambient dose equivalents, H(p)(10) and H(*)(10), by using a specially developed measuring system. The aim of this project was to determine the range of magnitudes of doses for representative personnel and cargo X-ray scanner systems. Depending on the type of scanner, the determined dose values for personnel scanners range from 0.07 microSv to 6 microSv. Measurements and instruments used in this study are described and the dose values obtained are discussed in detail. PMID- 16698968 TI - Pitfalls and modelling inconsistencies in computational radiation dosimetry: lessons learnt from the QUADOS intercomparison. Part I: Neutrons and uncertainties. AB - The QUADOS EU cost shared action conducted an intercomparison on the usage of numerical methods in radiation protection and dosimetry. The eight problems proposed were intended to test the usage of Monte Carlo and deterministic methods by assessing the accuracy with which the codes are applied and also the methods used to evaluate uncertainty in the answer gained through these methods. The overall objective was to spread good practice through the community and give users information on how to assess the uncertainties associated with their calculated results. PMID- 16698969 TI - Response of LiF:Mg,Ti to low energy carbon and oxygen ions. AB - The extended track interaction model (ETIM) has been formulated to explain the TL fluence response for peak 5 to heavy ions using radial dose distributions produced by the ions in LiF and their luminescent centre occupation probability distributions. In this work, an experimental study of the TLD-100 fluence response to carbon and oxygen ions and its interpretation in terms of a Monte Carlo simulation of ETIM applied to peak 5 are presented. Irradiations were performed with 7.34 and 10.3 MeV 12C and 8.34 MeV 16O ions in the fluence interval between 2 x 10(7) and 2 x 10(11) cm(-2). Individual glow curve responses show the expected increase of supralinearity as the peak temperature increases. Data for peak 5 show a weak dependence with energy. These latter results are difficult to understand when one considers the differences in the expected radial occupancies for different ion energies. PMID- 16698970 TI - Deformation luminescence produced during application and release of pressure on to gamma-irradiated CaF2:RE crystals. AB - Calcium fluoride CaF2 is an interesting host lattice for rare earth (RE) activators. CaF2 crystals doped with different concentrations of Dy, Ce, Er and Gd have been grown by the Bridgman technique and their deformation luminescence (DL) induced by room temperature gamma irradiation has been recorded. When a uniaxial pressure is applied on to gamma-irradiated CaF2:RE crystals, initially the DL intensity increases with time, attains a maximum value and then it decreases with time. Although the DL intensity produced during the release of pressure is less, its rise and decay behaviours are similar to that obtained during the application of pressure. The DL intensity depends on dopant, concentration of dopant, irradiation doses and mass of the load or applied pressure. It is suggested that the moving dislocation produced during deformation of crystals capture holes from hole trapped centres (like perturbed Vk centre) and the subsequent radiative recombination of the dislocation holes with electrons give rise to DL. PMID- 16698971 TI - Modelling the dose response of peaks 4, 5 and 5b, in TLD-100, as a function of recombination temperature. AB - This investigation focuses on whether the experimentally observed changes, in supralinearity f(D) as a function of recombination temperature, can be, successfully, predicted by the Unified Interaction Model (UNIM). To resolve this question, the measurement of f(D) has been carried out for glow peaks 4, 5 and 5b in LiF:Mg,Ti (TLD-100) at two different heating rates and UNIM analysis is used in an attempt to arrive at a consistent/physically reasonable explanation of all the f(D)/heating rate data for the three glow peaks. It is demonstrated that in addition to the variation in the luminescence and competitive centre capture cross section with temperature changes in the ks (the geminate recombination coefficient) parameter as a function of temperature is also required in order to predict the changes in f(D). PMID- 16698972 TI - Community health advocacy. AB - Competing health needs of diverse populations and ever shrinking resources available to support these needs often serve as the impetus for the initiation of advocacy efforts to improve community health. However, perceptions of what constitutes a community differ, as do approaches to advocacy itself. This glossary addresses five key questions: (1) What is advocacy?; (2) What is meant by community?; (3) What are the different approaches to community health advocacy?; (4) How are priorities established in the face of competing health advocacy goals?; (5) How can community health advocacy efforts be evaluated?; and (6) What challenges may be encountered in advocating for community health? Each of these issues could serve as the basis for a text on that subject alone. Accordingly, this article is not meant to be comprehensive text on these issues but is, instead, intended to highlight key foundational issues. And, although advocacy efforts can be conducted by individuals, this article focuses specifically on advocacy efforts of communities, however they may be defined and characterised. PMID- 16698973 TI - The legionnaires' outbreak in Barrow-in-Furness, summer 2002. PMID- 16698974 TI - Community acquired legionnaires' disease: lessons for surveillance from recent outbreaks. PMID- 16698975 TI - Living arrangements and mental health in Finland. AB - BACKGROUND: Non-married persons are known to have poor mental health compared with married persons. Health differences between marital status groups may largely arise from corresponding differences in interpersonal social bonds. However, official marital status mirrors the social reality of persons to a decreasing extent, and living arrangements may be a better measure of social bonds. Little is known about mental health in different living arrangement groups. This study aims to establish the extent and determinants of mental health differences by living arrangement in terms of psychological distress (GHQ) and DSM-IV psychiatric disorders (CIDI). METHODS: Data were used from the nationally representative cross sectional health 2000 survey, conducted in 2000-1 in Finland. Altogether 4685 participants (80%) aged 30-64 years were included in these analyses; comprehensive information was available on measures of mental health and living arrangements. Living arrangements were measured as follows: married, cohabiting, living with other(s) than a partner, and living alone. RESULTS: Compared with the married, persons living alone and those living with other(s) than a partner were approximately twice as likely to have anxiety or depressive disorders. Cohabiters did not differ from the married. In men, psychological distress was similarly associated with living arrangements. Unemployment, lack of social support, and alcohol consumption attenuated the excess psychological distress and psychiatric morbidity of persons living alone and of those living with other(s) than a partner by about 10%-50% each. CONCLUSIONS: Living arrangements are strongly associated with mental health, particularly among men. Information on living arrangements, social support, unemployment, and alcohol use may facilitate early stage recognition of poor mental health in primary health care. PMID- 16698976 TI - Do perceptions of neighbourhood environment influence health? Baseline findings from a British survey of aging. AB - OBJECTIVES: To investigate the relation between perceived neighbourhood environment, social contact and support, and self efficacy, on the health of older people. DESIGN: British cross sectional population survey of people aged 65 and over. PARTICIPANTS: 999 people aged 65 plus living at home in Britain. RESULTS: Regression modelling showed that high self efficacy had a strong independent association with better self rated health status and physical functioning. Indicators of perceived neighbourhood environment that showed strong associations with both good health and functioning were: perceptions of good quality facilities in the area (social/leisure, facilities for people aged 65+, rubbish collection, health services, transport, closeness to shops, somewhere nice to walk), and high levels of neighbourliness (knows/trusts people). Perceptions of problems in the area (noise, crime, air quality, rubbish/litter, traffic, graffiti) were also predictive of poorer health. Measures of social contact and support did not show any independent associations with health or functioning. CONCLUSIONS: The unique value of this paper is in the simultaneous analysis of associations between perceived neighbourhood, social contact and support, self efficacy, and health. The consistent strength of older people's perceptions of the quality of facilities in their neighbourhoods shows that responsive reinforcement of local infrastructures might have wider health benefits. Also of interest was the strength of self efficacy as a predictor of self rated health and physical functioning. The results have implications for both neighbourhood level interventions and self management programmes in chronic illness. PMID- 16698977 TI - Life course social roles and women's health in mid-life: causation or selection? AB - STUDY OBJECTIVE: To investigate whether relations between social roles and health are explained by health selection into employment and parenthood by examining the influence of early health on relations between long term social role histories and health in mid-life. DESIGN: Prospective, population based, birth cohort study. PARTICIPANTS AND SETTING: Women from a national British cohort born in 1946, including 1171 women with a valid measure of self reported health at age 54 and valid work and family role measures at ages 26, 36, 43, and 53, as well as 1433 women with a valid body mass index (BMI) measure at age 53 and valid work and family role measures at ages 26, 36, 43, and 53. OUTCOME MEASURES: Self reported health at age 54 and obesity at age 53, taken from objective height and weight measures conducted by a survey nurse during face to face interviews in respondents' homes. MAIN RESULTS: Women who occupied multiple roles over the long term reported relatively good health at age 54 and this was not explained by early health. Women with weak long term ties to the labour market were more likely to be obese at age 53. Examination of body mass index (BMI) from age 15 showed that long term homemakers were larger than other women from age 26, but their mean BMI increased significantly more with age than that of other women. CONCLUSIONS: Relations between social roles and health were generally not explained by health selection into employment and parenthood, although some health selection may occur for obesity. PMID- 16698978 TI - Does gender modify associations between self rated health and the social and economic characteristics of local environments? AB - OBJECTIVES: To examine whether area level socioeconomic disadvantage and social capital have different relations with women's and men's self rated health. METHODS: The study used data from 15 112 respondents to the 1998 Tasmanian (Australia) healthy communities study (60% response rate) nested within 41 statistical local areas. Gender stratified analyses were conducted of the associations between the index of relative socioeconomic disadvantage (IRSD) and social capital (neighbourhood integration, neighbourhood alienation, neighbourhood safety, political participation, social trust, trust in institutions) and individual level self rated health using multilevel logistic regression analysis before (age only) and after adjustment for individual level confounders (marital status, indigenous status, income, education, occupation, smoking). The study also tested for interactions between gender and area level variables. RESULTS: IRSD was associated with poor self rated health for women (age adjusted p<0.001) and men (age adjusted p<0.001), however, the estimates attenuated when adjusted for individual level variables. Political participation and neighbourhood safety were protective for women's self rated health but not for men's. Interactions between gender and political participation (p = 0.010) and neighbourhood safety (p = 0.023) were significant. CONCLUSIONS: These finding suggest that women may benefit more than men from higher levels of area social capital. PMID- 16698979 TI - Influence of marital history over two and three generations on early death. A longitudinal study of Danish men born in 1953. AB - BACKGROUND: This study examined the effects of marital status over two and three generations on last generation's mortality, and tested the hypothesis of an effect of the latest status (proximity effect) as well as the hypothesis of an accumulative effect. METHODS: The study population covers a random sample of all boys born in the the metropolitan area of Copenhagen with complete data from interviews and registers on two and three generation's marital status, socioeconomic position variables, and last generation's admission to psychiatric hospital, n = 2614. Among these 105 deaths occurred. Cox proportional hazards regression models were used to estimate the effect of marital status on mortality. RESULTS: Never married sons showed a considerably increased mortality compared with their married counterparts in the adjusted analyses. Mother's marital status at childbirth was also associated with increased mortality among the sons. There was no independent effect of maternal grandparent's experience of divorce on third generation's mortality. Son's marital status was the strongest marital status predictor of mortality. Accumulation of both two and three generations' marital status was significantly associated with mortality risk in a dose-response pattern. All analyses were adjusted for socioeconomic position variables and mental health. CONCLUSIONS: These results support the proximity hypothesis as son's marital status was the strongest predictor of mortality, and suggest an accumulative effect as each of the three non-married generations added to an increased mortality risk. PMID- 16698980 TI - Influence of family type and parenting behaviours on teenage sexual behaviour and conceptions. AB - BACKGROUND: Longitudinal data were used to explore relations between teenage pregnancy, sexual behaviour, and family type. The study examined whether students from lone parent and/or teenage mother initiated families more commonly report sex, lack of contraception at first sex, and/or conceptions by age 15/16, and whether such associations can be explained by low parental strictness, difficult parent-child communication, and/or low parental input into sex education. Up to date longitudinal UK research on family influences on conceptions is lacking, as is longitudinal research on family influences on sexual behaviour. No previous studies have comprehensively examined effects of parenting behaviours. Unlike previous research, this study tested theories suggesting that parenting deficits among lone parent and teenage initiated families increase risk of teenage pregnancy among their children. METHODS: Secondary analysis of data from a trial of sex education. RESULTS: Girls and boys from lone parent families or having mothers who were teenagers when they were born were more likely to report sex but not lack of contraception at first sex by age 15/16. Girls and boys with mothers having them as teenagers, and boys but not girls from lone parent families, were more likely to report being involved in conceptions by age 15/16. Only the association between teenage mother family and girls' conceptions was reduced by adjusting for a parenting behaviour measure. CONCLUSIONS: Students from lone parent families or having mothers who were teenagers when they were born are more likely to report early sexual debut and conceptions by age 15/16, but this is not generally explained by parenting style. PMID- 16698981 TI - Traffic intensity, dwelling value, and hospital admissions for respiratory disease among the elderly in Montreal (Canada): a case-control analysis. AB - BACKGROUND: Persons exposed to residential traffic have increased rates of respiratory morbidity and mortality. As poverty is an important determinant of ill health, some have argued that these associations may relate to the lower socioeconomic status of those living along major roads. AIMS: The objective was to evaluate the association between traffic intensity at home and hospital admissions for respiratory disease among Montreal residents of 60 years and older. METHODS: Case hospitalisations were those with respiratory diagnoses and control hospitalisations were those where the primary discharge diagnosis was non respiratory. Morning peak traffic estimates from the EMME/2 Montreal traffic model (MOTREM98) were used as an indicator of exposure to road traffic outside the homes of those hospitalised. The crude association between traffic intensity and hospitalisation for respiratory disease was adjusted by an area based estimate of the appraised value of patients' residences, expressed as a dollar average over a small segment of road (lodging value). This indicator of socioeconomic status, as calculated from the Montreal property assessment database, is available at a finer geographical scale than the neighbourhood socioeconomic indicators accessible from the Canadian census. RESULTS: Increased odds of being hospitalised for a respiratory compared with a control diagnosis were associated with higher levels of estimated road traffic nearby patients' homes, even after adjustment for lodging value (crude OR 1.35, CI95% 1.22 to 1.49; adjusted OR 1.18, CI95% 1.06 to 1.31 for >3160 vehicles passing during the three hour morning traffic peak compared with secondary roads off network). CONCLUSION: The results suggest that road traffic intensity itself, may affect the respiratory health of elderly residents of a large Canadian city, an association that is not solely a reflection of socioeconomic status. PMID- 16698983 TI - People's power in people's hands: the lesson we need to learn. PMID- 16698984 TI - Impact of a cultural belief about ghost month on delivery mode in Taiwan. AB - BACKGROUND: Many Chinese believe the lunar month of July, "ghost month" is inauspicious for major surgical procedures. This study hypothesised that caesaren delivery (CS) rates will be significantly lower during lunar July, and higher than normal during June, representing preemptive caesaren delivery to avoid delivering in July. METHODS: Population based data from Taiwan on all singleton deliveries during 1997-2003 (1 750 862 cases) were subjected to multivariate autoregressive integrated moving average (ARIMA) modelling, adjusting for major obstetric complications (previous CS, breech presentation, dystocia, and fetal distress). RESULTS: ARIMA intervention models showed significantly lower CS rates in lunar July, and among younger age groups (p<0.001), but not among 35 plus aged mothers. Incidence of previous CS, is significantly higher among June deliveries, while the incidence of the remaining major complications is similar in July, June, and other months. Patients with clinically less salient obstetric complications show significantly lower CS rates in July. CONCLUSIONS: Adjusted CS rates during the ghost month are significantly lower than other months. Lunar June shows an increase in deliveries of previous CS mothers (almost all by CS), suggesting elective CS to preempt CS in July. A major policy implication is that health education must be launched to dissipate the cultural belief about the ghost month. Evidence also implies some proportion of clinically un-indicated CS in other months, showing the need for professional and policy initiatives to reduce unnecessary CS. Policy makers and researchers in other countries should be alert to cultural beliefs associated with delivery to enable informed delivery choices by mothers. PMID- 16698982 TI - Income inequality and physical and mental health: testing associations consistent with proposed causal pathways. AB - OBJECTIVES: To test associations between individual health outcomes and ecological variables proposed in causal models of relations between income inequality and health. DESIGN: Regression analysis of a large, nationally representative dataset, linked to US census and other county and state level sources of data on ecological covariates. The regressions control for individual economic and demographic covariates as well as relevant potential ecological confounders. SETTING: The US population in the year 2000. PARTICIPANTS: 4817 US adults about age 40, representative of the US population. MAIN OUTCOME MEASURES: Two outcomes were studied: self reported general health status, dichotomised as "fair" or "poor" compared with "excellent", "very good", or "good", and depression as measured by a score on the Center for Epidemiologic Studies depression instrument >16. RESULTS: State generosity was significantly associated with a reduced odds of reporting poor general health (OR 0.84, 95%CI: 0.71 to 0.99), and the county unemployment rate with reduced odds of reporting depression (OR 0.91, 95%CI: 0.84 to 0.97). The measure of income inequality is a significant risk factor for reporting poor general health (OR 1.98, CI: 1.08 to 3.62), controlling for all ecological and individual covariates. In stratified models, the index of social capital is associated with reduced odds of reporting poor general health among black people and Hispanics (OR 0.40, CI: 0.18 to 0.90), but not significant among white people. The inequality measure is significantly associated with reporting poor general health among white people (OR 2.60, CI: 1.22 to 5.56) but not black people and Hispanics. CONCLUSIONS: The effect of income inequality on health may work through the influence of invidious social comparisons (particularly among white subjects) and (among black subjects and Latinos) through a reduction in social capital. Researchers may find it fruitful to recognise the cultural specificity of any such effects. PMID- 16698985 TI - Childhood socioeconomic circumstances predict specific causes of death in adulthood: the Glasgow student cohort study. PMID- 16698986 TI - Can Health Promoting Schools contribute to the better health and wellbeing of young people? The Hong Kong experience. AB - BACKGROUND: The Health Promoting School (HPS) is a WHO sponsored framework, compiled to enable education and health sectors to be more effective in school based initiatives. AIMS: This study attempted to test the hypothesis that students from schools that had comprehensively embraced the HPS concept as indicated by the Healthy School Award, were better, in terms of health risk behaviour, self reported health status, and academic results, than students from schools that did not reach the standard of the award. METHODS AND RESULTS: The results presented came from nine schools (four primary and five secondary) applying for accreditation of the Healthy Schools Award after adopting the HPS framework for two years. Regular consultancy support and training were available to all schools. Students had completed before and after surveys to assess their health behaviours, self reported health status, and academic standing before the two year intervention, and at its end. Data from the before and after surveys of the students attending schools that reached certain level of HPS standard as indicated by the award, were compared with students whose schools did not receive the award, and the results showed differences. Some differences were found to be more significant among the primary school students than secondary schools students. This illustrated early intervention for lifestyle changes to be more effective. Students' satisfaction with life also improved if their schools adopted the concept of HPS comprehensively. CONCLUSIONS: The results suggest that comprehensive implementation of HPS would contribute to differences in certain behaviours and self reported health and academic status. PMID- 16698987 TI - Injecting and HIV prevalence among young heroin users in three Spanish cities and their association with the delayed implementation of harm reduction programmes. AB - OBJECTIVES: To evaluate changes in the prevalence of HIV infection among young heroin users in three Spanish cities, and their association with harm reduction programmes (HRPs). METHODS: Two cross sectional studies. The 1995 study included 596 users; half were street recruited and half were recruited at drug treatment centres. The 2001-03 study included 981 street recruited users. Face to face interviews were conducted using a structured questionnaire. Samples for HIV testing (saliva in 1995 and dried blood spot in 2001-03) were collected. RESULTS: The proportion who had ever injected (IDUs) decreased in all three cities. HIV prevalence in IDUs decreased by half in Barcelona (44.1% to 20.8%) and Seville (44.2% to 22.2%), but remained constant in Madrid (36.8% and 34.9%). This difference was attributable to a decrease in HIV prevalence in long term IDUs in Barcelona and Seville, but not in Madrid. The crude odds ratio for HIV prevalence in Madrid compared with Barcelona in long term IDUs was 2.3 (95%CI 1.4 to 3.7), increasing to 3.1 (95%CI 1.5 to 6.2) after adjusting for sociodemographic and risk factors. HIV prevalence in short term IDUs was similar in all cities. In 1992 Barcelona already had 20 heroin users in methadone maintenance programmes (MMPs) per 10 000 population aged 15-49 years; Seville reached this rate in 1994, and Madrid, not until 1998. CONCLUSIONS: The prevalence of HIV infection did not decrease in long term injectors in Madrid. The delayed implementation of HRPs, especially MMPs, may be the most plausible hypothesis. This finding should shed light on decision making in countries in a similar epidemiological and sociological situation. PMID- 16698988 TI - Review of syndromic surveillance: implications for waterborne disease detection. AB - Syndromic surveillance is the gathering of data for public health purposes before laboratory or clinically confirmed information is available. Interest in syndromic surveillance has increased because of concerns about bioterrorism. In addition to bioterrorism detection, syndromic surveillance may be suited to detecting waterborne disease outbreaks. Theoretical benefits of syndromic surveillance include potential timeliness, increased response capacity, ability to establish baseline disease burdens, and ability to delineate the geographical reach of an outbreak. This review summarises the evidence gathered from retrospective, prospective, and simulation studies to assess the efficacy of syndromic surveillance for waterborne disease detection. There is little evidence that syndromic surveillance mitigates the effects of disease outbreaks through earlier detection and response. Syndromic surveillance should not be implemented at the expense of traditional disease surveillance, and should not be relied upon as a principal outbreak detection tool. The utility of syndromic surveillance is dependent on alarm thresholds that can be evaluated in practice. Syndromic data sources such as over the counter drug sales for detection of waterborne outbreaks should be further evaluated. PMID- 16698989 TI - On socioeconomic gradients in cancer registry data quality. PMID- 16698991 TI - Murine norovirus: a model system to study norovirus biology and pathogenesis. PMID- 16698992 TI - Inhibition of the NF-kappaB pathway by varicella-zoster virus in vitro and in human epidermal cells in vivo. AB - Varicella-zoster virus (VZV) is an alphaherpesvirus that causes varicella and herpes zoster. Using human cellular DNA microarrays, we found that many nuclear factor kappa B (NF-kappaB)-responsive genes were down-regulated in VZV-infected fibroblasts, suggesting that VZV infection inhibited the NF-kappaB pathway. The activation of this pathway causes a cellular antiviral response, including the production of alpha/beta interferon, cytokines, and other proteins that restrict viral infection. In these experiments, we demonstrated that VZV interferes with NF-kappaB activation in cultured fibroblasts and in differentiated epidermal cells in skin xenografts of SCIDhu mice infected in vivo. VZV infection of fibroblasts caused a transient nuclear translocation of p50 and p65, the canonical NF-kappaB family members. In a process that was dependent upon the presence of infectious VZV, these proteins rapidly became sequestered in the cytoplasm of VZV-infected cells. Exclusion of NF-kappaB proteins from nuclei was associated with the continued presence of IkappaBalpha, which binds p50 and p65 and prevents their nuclear accumulation. IkappaBalpha levels did not diminish even though the protein became phosphorylated and ubiquitinated, as determined based on detection of the characteristic high-molecular-weight form of the protein, and the 26S proteasome remained functional in VZV-infected cells. VZV infection also inhibited the characteristic degradation of IkappaBalpha that is induced by exposure of fibroblasts to tumor necrosis factor alpha. As expected, herpes simplex virus 1 caused the persistent nuclear translocation of NF-kappaB proteins, which has been shown to facilitate its replication, whereas VZV infection progressed without persistent NF-kappaB nuclear localization. We suggest that VZV has evolved a mechanism to limit host cell antiviral defenses by sequestering NF-kappaB proteins in the cytoplasm, a strategy that appears to be unique among the herpesviruses. PMID- 16698993 TI - Epstein-Barr virus protein kinase BGLF4 is a virion tegument protein that dissociates from virions in a phosphorylation-dependent process and phosphorylates the viral immediate-early protein BZLF1. AB - Epstein-Barr virus (EBV) BGLF4 is a viral protein kinase that is expressed in the lytic phase of infection and is packaged in virions. We report here that BGLF4 is a tegument protein that dissociates from the virion in a phosphorylation dependent process. We also present evidence that BGLF4 interacts with and phosphorylates BZLF1, a key viral regulator of lytic infection. These conclusions are based on the following observations. (i) In in vitro tegument release assays, a significant fraction of BGLF4 was released from virions in the presence of physiological NaCl concentrations. (ii) Addition of physiological concentrations of ATP and MgCl(2) to virions enhanced BGLF4 release, but phosphatase treatment of virions significantly reduced BGLF4 release. (iii) A recombinant protein containing a domain of BZLF1 was specifically phosphorylated by purified recombinant BGLF4 in vitro, and BGLF4 altered BZLF1 posttranslational modification in vivo. (iv) BZLF1 was specifically coimmunoprecipitated with BGLF4 in 12-O-tetradecanoylphorbol-13-acetate-treated B95-8 cells and in COS-1 cells transiently expressing both of these viral proteins. (v) BGLF4 and BZLF1 were colocalized in intranuclear globular structures, resembling the viral replication compartment, in Akata cells treated with anti-human immunoglobulin G. Our results suggest that BGLF4 functions not only in lytically infected cells by phosphorylating viral and cellular targets but also immediately after viral penetration like other herpesvirus tegument proteins. PMID- 16698994 TI - Ebola virus VP35-VP40 interaction is sufficient for packaging 3E-5E minigenome RNA into virus-like particles. AB - The packaging of viral genomic RNA into nucleocapsids and subsequently into virions is not completely understood. Phosphoprotein (P) and nucleoprotein (NP) interactions link NP-RNA complexes with P-L (polymerase) complexes to form viral nucleocapsids. The nucleocapsid then interacts with the viral matrix protein, leading to specific packaging of the nucleocapsid into the virion. A mammalian two-hybrid assay and confocal microscopy were used to demonstrate that Ebola virus VP35 and VP40 interact and colocalize in transfected cells. VP35 was packaged into budding virus-like particles (VLPs) as observed by protease protection assays. Moreover, VP40 and VP35 were sufficient for packaging an Ebola virus minigenome RNA into VLPs. Results from immunoprecipitation-reverse transcriptase PCR experiments suggest that VP35 confers specificity of the nucleocapsid for viral genomic RNA by direct VP35-RNA interactions. PMID- 16698995 TI - Differentially regulated interferon response determines the outcome of Newcastle disease virus infection in normal and tumor cell lines. AB - Newcastle disease virus (NDV) is a negative-strand RNA virus with oncolytic activity against human tumors. Its effectiveness against tumors and safety in normal tissue have been demonstrated in several clinical studies. Here we show that the spread of NDV infection is drastically different in normal cell lines than in tumor cell lines and that the two cell types respond differently to beta interferon (IFN-beta) treatment. NDV rapidly replicated and killed HT-1080 human fibrosarcoma cells but spread poorly in CCD-1122Sk human skin fibroblast cells. Pretreatment with endogenous or exogenous IFN-beta completely inhibited NDV replication in normal cells but had little or no effect in tumor cells. Thus, the outcome of NDV infection appeared to depend on the response of uninfected cells to IFN-beta. To investigate their differences in IFN responsiveness, we analyzed and compared the expression and activation of components of the IFN signal transduction pathway in these two types of cells. The levels of phosphorylated STAT1 and STAT2 and that of the ISGF3 complex were markedly reduced in IFN-beta treated tumor cells. Moreover, cDNA microarray analysis revealed significantly fewer IFN-regulated genes in the HT-1080 cells than in the CDD-1122Sk cells. This finding suggests that tumor cells demonstrate a less-than-optimum antiviral response because of a lesion in their IFN signal transduction pathway. The rapid spread of NDV in HT-1080 cells appears to be caused by their deficient expression of anti-NDV proteins upon exposure to IFN-beta. PMID- 16698996 TI - Ebola virus VP24 binds karyopherin alpha1 and blocks STAT1 nuclear accumulation. AB - Ebola virus (EBOV) infection blocks cellular production of alpha/beta interferon (IFN-alpha/beta) and the ability of cells to respond to IFN-alpha/beta or IFN gamma. The EBOV VP35 protein has previously been identified as an EBOV-encoded inhibitor of IFN-alpha/beta production. However, the mechanism by which EBOV infection inhibits responses to IFNs has not previously been defined. Here we demonstrate that the EBOV VP24 protein functions as an inhibitor of IFN alpha/beta and IFN-gamma signaling. Expression of VP24 results in an inhibition of IFN-induced gene expression and an inability of IFNs to induce an antiviral state. The VP24-mediated inhibition of cellular responses to IFNs correlates with the impaired nuclear accumulation of tyrosine-phosphorylated STAT1 (PY-STAT1), a key step in both IFN-alpha/beta and IFN-gamma signaling. Consistent with this proposed function for VP24, infection of cells with EBOV also confers a block to the IFN-induced nuclear accumulation of PY-STAT1. Further, VP24 is found to specifically interact with karyopherin alpha1, the nuclear localization signal receptor for PY-STAT1, but not with karyopherin alpha2, alpha3, or alpha4. Overexpression of VP24 results in a loss of karyopherin alpha1-PY-STAT1 interaction, indicating that the VP24-karyopherin alpha1 interaction contributes to the block to IFN signaling. These data suggest that VP24 is likely to be an important virulence determinant that allows EBOV to evade the antiviral effects of IFNs. PMID- 16698997 TI - Ebola virus VP35 protein binds double-stranded RNA and inhibits alpha/beta interferon production induced by RIG-I signaling. AB - The Ebola virus (EBOV) VP35 protein blocks the virus-induced phosphorylation and activation of interferon regulatory factor 3 (IRF-3), a transcription factor critical for the induction of alpha/beta interferon (IFN-alpha/beta) expression. However, the mechanism(s) by which this blockage occurs remains incompletely defined. We now provide evidence that VP35 possesses double-stranded RNA (dsRNA) binding activity. Specifically, VP35 bound to poly(rI) . poly(rC)-coated Sepharose beads but not control beads. In contrast, two VP35 point mutants, R312A and K309A, were found to be greatly impaired in their dsRNA-binding activity. Competition assays showed that VP35 interacted specifically with poly(rI) . poly(rC), poly(rA) . poly(rU), or in vitro-transcribed dsRNAs derived from EBOV sequences, and not with single-stranded RNAs (ssRNAs) or double-stranded DNA. We then screened wild-type and mutant VP35s for their ability to target different components of the signaling pathways that activate IRF-3. These experiments indicate that VP35 blocks activation of IRF-3 induced by overexpression of RIG-I, a cellular helicase recently implicated in the activation of IRF-3 by either virus or dsRNA. Interestingly, the VP35 mutants impaired for dsRNA binding have a decreased but measurable IFN antagonist activity in these assays. Additionally, wild-type and dsRNA-binding-mutant VP35s were found to have equivalent abilities to inhibit activation of the IFN-beta promoter induced by overexpression of IPS 1, a recently identified signaling molecule downstream of RIG-I, or by overexpression of the IRF-3 kinases IKKepsilon and TBK-1. These data support the hypothesis that dsRNA binding may contribute to VP35 IFN antagonist function. However, additional mechanisms of inhibition, at a point proximal to the IRF-3 kinases, most likely also exist. PMID- 16698998 TI - LC16m8, a highly attenuated vaccinia virus vaccine lacking expression of the membrane protein B5R, protects monkeys from monkeypox. AB - The potential threat of smallpox as a bioweapon has led to the production and stockpiling of smallpox vaccine in some countries. Human monkeypox, a rare but important viral zoonosis endemic to central and western Africa, has recently emerged in the United States. Thus, even though smallpox has been eradicated, a vaccinia virus vaccine that can induce protective immunity against smallpox and monkeypox is still invaluable. The ability of the highly attenuated vaccinia virus vaccine strain LC16m8, with a mutation in the important immunogenic membrane protein B5R, to induce protective immunity against monkeypox in nonhuman primates was evaluated in comparison with the parental Lister strain. Monkeys were immunized with LC16m8 or Lister and then infected intranasally or subcutaneously with monkeypox virus strain Liberia or Zr-599, respectively. Immunized monkeys showed no symptoms of monkeypox in the intranasal-inoculation model, while nonimmunized controls showed typical symptoms. In the subcutaneous inoculation model, monkeys immunized with LC16m8 showed no symptoms of monkeypox except for a mild ulcer at the site of monkeypox virus inoculation, and those immunized with Lister showed no symptoms of monkeypox, while nonimmunized controls showed lethal and typical symptoms. These results indicate that LC16m8 prevents lethal monkeypox in monkeys, and they suggest that LC16m8 may induce protective immunity against smallpox. PMID- 16698999 TI - Role of the stable signal peptide and cytoplasmic domain of G2 in regulating intracellular transport of the Junin virus envelope glycoprotein complex. AB - Enveloped viruses utilize the membranous compartments of the host cell for the assembly and budding of new virion particles. In this report, we have investigated the biogenesis and trafficking of the envelope glycoprotein (GP-C) of the Junin arenavirus. The mature GP-C complex is unusual in that it retains a stable signal peptide (SSP) as an essential component in association with the typical receptor-binding (G1) and transmembrane fusion (G2) subunits. We demonstrate that, in the absence of SSP, the G1-G2 precursor is restricted to the endoplasmic reticulum (ER). This constraint is relieved by coexpression of SSP in trans, allowing transit of the assembled GP-C complex through the Golgi and to the cell surface, the site of arenavirus budding. Transport of a chimeric CD4 glycoprotein bearing the transmembrane and cytoplasmic domains of G2 is similarly regulated by SSP association. Truncations to the cytoplasmic domain of G2 abrogate SSP association yet now permit transport of the G1-G2 precursor to the cell surface. Thus, the cytoplasmic domain of G2 is an important determinant for both ER localization and its control through SSP binding. Alanine mutations to either of two dibasic amino acid motifs in the G2 cytoplasmic domain can also mobilize the G1-G2 precursor for transit through the Golgi. Taken together, our results suggest that SSP binding masks endogenous ER localization signals in the cytoplasmic domain of G2 to ensure that only the fully assembled, tripartite GP-C complex is transported for virion assembly. This quality control process points to an important role of SSP in the structure and function of the arenavirus envelope glycoprotein. PMID- 16699001 TI - Evidence for potent autologous neutralizing antibody titers and compact envelopes in early infection with subtype C human immunodeficiency virus type 1. AB - Information about neutralizing antibody responses in subtype C-infected individuals is limited, even though this viral subtype causes the majority of AIDS cases worldwide. Here we compared the course and magnitude of the autologous neutralizing antibody (NAb) response against viral envelope (Env) glycoproteins present during acute and early infection with subtypes B and C human immunodeficiency virus type 1 (HIV-1). NAb responses were evaluated in 6 subtype B-infected and 11 subtype C-infected subjects over a mean evaluation period of 25 months using a pseudovirus reporter gene assay. All subjects in the C cohort were infected through heterosexual contact, while five of the six subjects in the B cohort were infected via male-to-male contact. The kinetics and magnitude of the NAb responses varied among subjects in the B and C cohorts; however, the median 50% inhibitory concentration (IC(50) titer) reached by antibody in the plasma of subtype C-infected subjects, overall, was 3.5-fold higher than in the subtype B infected subjects (P = 0.06). The higher titers of NAbs in the C cohort were associated with viruses having significantly shorter amino acid length (P = 0.002) in the V1 to V4 region of the surface Env glycoprotein, gp120, compared to the B cohort. Despite the potency of the autologous subtype C NAb response, it was not directed against cross-neutralizing epitopes. These data demonstrate that subtype C Envs elicit a potent yet restricted NAb response early in infection that frequently reaches IC(50) titers in excess of 1:1,000 and suggest that clade specific differences may exist in Env immunogenicity or susceptibility to neutralization. PMID- 16699000 TI - Separate basic region motifs within the adeno-associated virus capsid proteins are essential for infectivity and assembly. AB - Adeno-associated virus (AAV) is gaining momentum as a gene therapy vector for human applications. However, there remain impediments to the development of this virus as a vector. One of these is the incomplete understanding of the biology of the virus, including nuclear targeting of the incoming virion during initial infection, as well as assembly of progeny virions from structural components in the nucleus. Toward this end, we have identified four basic regions (BR) on the AAV2 capsid that represent possible nuclear localization sequence (NLS) motifs. Mutagenesis of BR1 ((120)QAKKRVL(126)) and BR2 ((140)PGKKRPV(146)) had minor effects on viral infectivity ( approximately 4- and approximately 10-fold, respectively), whereas BR3 ((166)PARKRLN(172)) and BR4 ((307)RPKRLN(312)) were found to be essential for infectivity and virion assembly, respectively. Mutagenesis of BR3, which is located in Vp1 and Vp2 capsid proteins, does not interfere with viral production or trafficking of intact AAV capsids to the nuclear periphery but does inhibit transfer of encapsidated DNA into the nucleus. Substitution of the canine parvovirus NLS rescued the BR3 mutant to wild-type (wt) levels, supporting the role of an AAV NLS motif. In addition, rAAV2 containing a mutant form of BR3 in Vp1 and a wt BR3 in Vp2 was found to be infectious, suggesting that the function of BR3 is redundant between Vp1 and Vp2 and that Vp2 may play a role in infectivity. Mutagenesis of BR4 was found to inhibit virion assembly in the nucleus of transfected cells. This affect was not completely due to the inefficient nuclear import of capsid subunits based on Western blot analysis. In fact, aberrant capsid foci were observed in the cytoplasm of transfected cells, compared to the wild type, suggesting a defect in early viral assembly or trafficking. Using three-dimensional structural analysis, the lysine- and arginine-to-asparagine change disrupts hydrogen bonding between these basic residues and adjacent beta strand glutamine residues that may prevent assembly of intact virions. Taken together, these data support that the BR4 domain is essential for virion assembly. Each BR was also found to be conserved in serotypes 1 to 11, suggesting that these regions are significant and function similarly in each serotype. This study establishes the importance of two BR motifs on the AAV2 capsid that are essential for infectivity and virion assembly. PMID- 16699003 TI - The cytoplasmic tail of the influenza A virus M2 protein plays a role in viral assembly. AB - The viral replication cycle concludes with the assembly of viral components to form progeny virions. For influenza A viruses, the matrix M1 protein and two membrane integral glycoproteins, hemagglutinin and neuraminidase, function cooperatively in this process. Here, we asked whether another membrane protein, the M2 protein, plays a role in virus assembly. The M2 protein, comprising 97 amino acids, possesses the longest cytoplasmic tail (54 residues) of the three transmembrane proteins of influenza A viruses. We therefore generated a series of deletion mutants of the M2 cytoplasmic tail by reverse genetics. We found that mutants in which more than 22 amino acids were deleted from the carboxyl terminus of the M2 tail were viable but grew less efficiently than did the wild-type virus. An analysis of the virions suggested that viruses with M2 tail deletions of more than 22 carboxy-terminal residues apparently contained less viral ribonucleoprotein complex than did the wild-type virus. These M2 tail mutants also differ from the wild-type virus in their morphology: while the wild-type virus is spherical, some of the mutants were filamentous. Alanine-scanning experiments further indicated that amino acids at positions 74 to 79 of the M2 tail play a role in virion morphogenesis and affect viral infectivity. We conclude that the M2 cytoplasmic domain of influenza A viruses plays an important role in viral assembly and morphogenesis. PMID- 16699002 TI - Extraintestinal spread and replication of a homologous EC rotavirus strain and a heterologous rhesus rotavirus in BALB/c mice. AB - Although rotavirus infection has generally been felt to be restricted to the gastrointestinal tract, over the last two decades there have been sporadic reports of children with acute or fatal cases of rotavirus gastroenteritis testing positive for rotavirus antigen and/or nucleic acid in various extraintestinal locations such as serum, liver, kidney, bladder, testes, nasal secretions, cerebrospinal fluid, and the central nervous system. Recently, studies in animals and people have demonstrated that rotavirus antigenemia is a common event during natural infection. In this study, we extend these observations and compare the intestinal and extraintestinal spread of wild-type homologous murine rotavirus EC and a heterologous strain, rhesus rotavirus (RRV), in newborn mice. A strand-specific quantitative reverse transcription-PCR (ssQRT PCR) assay was used to quantify the ability of different rotavirus strains to spread and replicate extraintestinally. Both strain EC and RRV were detected extraintestinally in the mesenteric lymph nodes (MLN), livers, lungs, blood, and kidneys. Extraintestinal replication, as measured by ssQRT-PCR, was most prominent in the MLN and occurred to a lesser degree in the livers, kidneys, and lungs. In the MLN, strain EC and RRV had similar (P < 0.05) RNA copy numbers, although EC was present at a 10,000-fold excess over RRV in the small intestine. Rotavirus nonstructural protein 4 (NSP4) and/or assembled triple-layered particles, indicated by immunostaining with the VP7 conformation-dependent monoclonal antibody 159, were detected in the MLN, lungs, and livers of EC- and RRV-inoculated mice, confirming the ssQRT-PCR findings. Infectious RRV was detected in the MLN in quantities exceeding the amount present in the small intestines or blood. The cells in the MLN that supported rotavirus replication included dendritic cells and potentially B cells and macrophages. These data indicate that extraintestinal spread and replication occurs commonly during homologous and some heterologous rotaviral infections; that the substantial host range restrictions for rhesus rotavirus, a heterologous strain present in the intestine, are not necessarily apparent at systemic sites; that the level and location of extraintestinal replication varies between strains; that replication can occur in several leukocytes subsets; and that extraintestinal replication is likely a part of the normal pathogenic sequence of homologous rotavirus infection. PMID- 16699004 TI - Role of the herpes simplex virus helicase-primase complex during adeno-associated virus DNA replication. AB - A subset of DNA replication proteins of herpes simplex virus (HSV) comprising the single-strand DNA-binding protein, ICP8 (UL29), and the helicase-primase complex (UL5, UL8, and UL52 proteins) has previously been shown to be sufficient for the replication of adeno-associated virus (AAV). We recently demonstrated complex formation between ICP8, AAV Rep78, and the single-stranded DNA AAV genome, both in vitro and in the nuclear HSV replication domains of coinfected cells. In this study the functional role(s) of HSV helicase and primase during AAV DNA replication were analyzed. To differentiate between their necessity as structural components of the HSV replication complex or as active enzymes, point mutations within the helicase and primase catalytic domains were analyzed. In two complementary approaches the remaining HSV helper functions were either provided by infection with HSV mutants or by plasmid transfection. We show here that upon cotransfection of the minimal four HSV proteins (i.e., the four proteins constituting the minimal requirements for basal AAV replication), UL52 primase catalytic activity was not required for AAV DNA replication. In contrast, UL5 helicase activity was necessary for fully efficient replication. Confocal microscopy confirmed that all mutants retained the ability to support formation of ICP8-positive nuclear replication foci, to which AAV Rep78 colocalized in a manner strictly dependent on the presence of AAV single-stranded DNA (ssDNA). The data indicate that recruitment of AAV Rep78 and ssDNA to nuclear replication sites by the four HSV helper proteins is maintained in the absence of catalytic primase or helicase activities and suggest an involvement of the HSV UL5 helicase activity during AAV DNA replication. PMID- 16699005 TI - Kaposi's sarcoma-associated herpesvirus lytic gene ORF57 is essential for infectious virion production. AB - The ORF57 gene of Kaposi's sarcoma-associated herpesvirus (KSHV) encodes a nuclear protein expressed during the lytic phase of KSHV replication. An ORF57 homolog is present in all known human herpesviruses and many animal herpesviruses. Many of these proteins have been demonstrated to have essential transcriptional and posttranscriptional regulatory functions. ORF57 enhances expression of reporter genes posttranscriptionally in vitro and may synergize with transcription factors to enhance gene transcription. However, the biologic role of ORF57 in KSHV replication has not been established. In this study, we demonstrate that ORF57 is essential for productive KSHV lytic replication by constructing a recombinant KSHV in which ORF57 expression has been specifically inactivated. The ORF57-null KSHV recombinant was unable to produce virion progeny or fully express several other lytic KSHV genes except when ORF57 was provided in trans. The Epstein-Barr virus (EBV) homolog of ORF57, SM, was unable to rescue lytic KSHV virion production, although EBV SM does enhance KSHV lytic gene expression from the ORF57-null mutant. Conversely, ORF57 did not rescue an SM null recombinant EBV, indicating the existence of virus-specific functions for the ORF57 family of genes. PMID- 16699006 TI - Regulation of the EBNA1 Epstein-Barr virus protein by serine phosphorylation and arginine methylation. AB - The Epstein-Barr virus (EBV) EBNA1 protein is important for the replication and mitotic segregation of EBV genomes in latently infected cells and also activates the transcription of some of the viral latency genes. A Gly-Arg-rich region between amino acids 325 and 376 is required for both the segregation and transcriptional activation functions of EBNA1. Here we show that this region is modified by both arginine methylation and serine phosphorylation. Mutagenesis of the four potentially phosphorylated serines in this region indicated that phosphorylation of multiple serines contributes to the efficient segregation of EBV-based plasmids by EBNA1, at least in part by increasing EBNA1 binding to hEBP2. EBNA1 was also found to bind the arginine methyltransferases PRMT1 and PRMT5. Multiple arginines in the 325-376 region were methylated in vitro by PRMT1 and PRMT5, as was an N-terminal Gly-Arg-rich region between amino acids 41 and 50. EBNA1 was also shown to be methylated in vivo, predominantly in the 325-376 region. Treatment of cells with a methylation inhibitor or down-regulation of PRMT1 altered EBNA1 localization, resulting in the formation of EBNA1 rings around the nucleoli. The results indicate that EBNA1 function is influenced by both serine phosphorylation and arginine methylation. PMID- 16699007 TI - Acetylation of the latency-associated nuclear antigen regulates repression of Kaposi's sarcoma-associated herpesvirus lytic transcription. AB - Reactivation of the Kaposi's sarcoma-associated herpesvirus (KSHV) lytic cycle can be initiated by transcription activation of the ORF50 immediate early gene (Rta). We show that ORF50 transcription is actively repressed by the KSHV latency associated nuclear antigen (LANA) during latency. Depletion of LANA by small interfering RNA derepressed ORF50 transcription in the latently infected BCBL1 pleural effusion lymphoma-derived cell line. In contrast, overexpression of LANA suppressed ORF50 mRNA levels in BCBL1 cells. ORF50 transcription was significantly elevated during primary infection with recombinant virus lacking LANA, further indicating that LANA plays a role in lytic gene silencing during the establishment of latency. Chromatin immunoprecipitation assays indicated that LANA interacts with the ORF50 promoter region in latently infected cells. Histone deacetylase inhibitors, including sodium butyrate (NaB) and trichostatin A, caused the rapid dissociation of LANA from the ORF50 promoter. NaB treatment of latently infected BCBL1 cells disrupted a stable interaction between LANA and the cellular proteins Sp1 and histone H2B. We also found immunological and radiochemical evidence that LANA is subject to lysine acetylation after NaB treatment. These findings support the role of LANA as a transcriptional repressor of lytic reactivation and provide evidence that lysine acetylation regulates LANA interactions with chromatin, Sp1, and ORF50 promoter DNA. PMID- 16699008 TI - Mucosal administration of CpG oligodeoxynucleotide elicits strong CC and CXC chemokine responses in the vagina and serves as a potent Th1-tilting adjuvant for recombinant gD2 protein vaccination against genital herpes. AB - Although sexually transmitted pathogens are capable of inducing pathogen-specific immune responses, vaginal administration of nonreplicating antigens elicits only weak, nondisseminating immune responses. The present study was undertaken to examine the potential of CpG-containing oligodeoxynucleotide (CpG ODN) for induction of chemokine responses in the genital tract mucosa and also as a vaginal adjuvant in combination with glycoprotein D of herpes simplex virus type 2 (HSV-2) for induction of antigen-specific immune responses. We found that a single intravaginal administration of CpG ODN in mice stimulates a rapid and potent response of CC chemokines macrophage inflammatory protein 1alpha (MIP 1alpha), MIP-1beta, and RANTES as well as of CXC chemokines MIP-2 and IP-10 in the vagina and/or the genital lymph nodes. Importantly, intravaginal vaccination with recombinant gD2 in combination with CpG ODN gave rise to a strong antigen specific Th1-like immune response in the genital lymph nodes as well as the spleens of the vaccinated mice. Further, such an immunization scheme conferred both systemic and mucosal immunoglobulin G antibody responses as well as protection against an otherwise lethal vaginal challenge with HSV-2. These results illustrate the potential of CpG ODN for induction of potent chemokine responses in the genital tract and also as a vaginal adjuvant for generation of Th1-type mucosal and systemic immune responses towards a nonreplicating antigen derived from a sexually transmitted pathogen. These data have implications for the development of a mucosal vaccine against genital herpes and possibly other sexually transmitted diseases. PMID- 16699009 TI - Gag regulates association of human immunodeficiency virus type 1 envelope with detergent-resistant membranes. AB - Assembly of the human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein on budding virus particles is important for efficient infection of target cells. In infected cells, lipid rafts have been proposed to form platforms for virus assembly and budding. Gag precursors partly associate with detergent-resistant membranes (DRMs) that are believed to represent lipid rafts. The cytoplasmic domain of the envelope gp41 usually carries palmitate groups that were also reported to confer DRM association. Gag precursors confer budding and carry envelope glycoproteins onto virions via specific Gag-envelope interactions. Thus, specific mutations in both the matrix domain of the Gag precursor and gp41 cytoplasmic domain abrogate envelope incorporation onto virions. Here, we show that HIV-1 envelope association with DRMs is directly influenced by its interaction with Gag. Thus, in the absence of Gag, envelope fails to associate with DRMs. A mutation in the p17 matrix (L30E) domain in Gag (Gag L30E) that abrogates envelope incorporation onto virions also eliminated envelope association with DRMs in 293T cells and in the T-cell line, MOLT 4. These observations are consistent with a requirement for an Env-Gag interaction for raft association and subsequent assembly onto virions. In addition to this observation, we found that mutations in the gp41 cytoplasmic domain that abrogated envelope incorporation onto virions and impaired infectivity of cell free virus also eliminated envelope association with DRMs. On the basis of these observations, we propose that Gag-envelope interaction is essential for efficient envelope association with DRMs, which in turn is essential for envelope budding and assembly onto virus particles. PMID- 16699010 TI - Human papillomavirus type 16 E6 activates NF-kappaB, induces cIAP-2 expression, and protects against apoptosis in a PDZ binding motif-dependent manner. AB - Infection with human papillomavirus (HPV) is a critical factor in the pathogenesis of most cervical cancers and some aerodigestive cancers. The HPV E6 oncoprotein from high-risk HPV types contributes to the immortalization and transformation of cells by multiple mechanisms, including degradation of p53, transcriptional activation of human telomerase reverse transcriptase (hTERT), and degradation of several proteins containing PDZ domains. The ability of E6 to bind PDZ domain-containing proteins is independent of p53 degradation or hTERT activation but does correlate with oncogenic potential (R. A. Watson, M. Thomas, L. Banks, and S. Roberts, J. Cell Sci. 116:4925-4934, 2003) and is essential for induction of epithelial hyperplasia in vivo (M. L. Nguyen, M. M. Nguyen, D. Lee, A. E. Griep, and P. F. Lambert, J. Virol. 77:6957-6964, 2003). In this study, we found that HPV type 16 E6 was able to activate NF-kappaB in airway epithelial cells through the induction of nuclear binding activity of p52-containing NF kappaB complexes in a PDZ binding motif-dependent manner. Transcript accumulation for the NF-kappaB-responsive antiapoptotic gene encoding cIAP-2 and binding of nuclear factors to the proximal NF-kappaB binding site of the cIAP-2 gene promoter are induced by E6 expression. Furthermore, E6 is able to protect cells from TNF-induced apoptosis. All of these E6-dependent phenotypes are dependent on the presence of the PDZ binding motif of E6. Our results imply a role for targeting of PDZ proteins by E6 in NF-kappaB activation and protection from apoptosis in airway epithelial cells. PMID- 16699011 TI - Characterization of the early steps of hepatitis C virus infection by using luciferase reporter viruses. AB - The lack of an efficient system to produce hepatitis C virus (HCV) particles has impeded the analysis of the HCV life cycle. Recently, we along with others demonstrated that transfection of Huh7 hepatoma cells with a novel HCV isolate (JFH1) yields infectious viruses. To facilitate studies of HCV replication, we generated JFH1-based bicistronic luciferase reporter virus genomes. We found that RNA replication of the reporter construct was only slightly attenuated and that virus titers produced were only three- to fivefold lower compared to the parental virus, making these reporter viruses an ideal tool for quantitative analyses of HCV infections. To expand the scope of the system, we created two chimeric JFH1 luciferase reporter viruses with structural proteins from the Con1 (genotype 1b) and J6CF (genotype 2a) strains. Using these and the authentic JFH1 reporter viruses, we analyzed the early steps of the HCV life cycle. Our data show that the mode of virus entry is conserved between these isolates and involves CD81 as a key receptor for pH-dependent virus entry. Competition studies and time course experiments suggest that interactions of HCV with cell surface-resident glycosaminoglycans aid in efficient infection of Huh7 cells and that CD81 acts during a postattachment step. The reporter viruses described here should be instrumental for investigating the viral life cycle and for the development of HCV inhibitors. PMID- 16699012 TI - A novel assay for viral microRNA function identifies a single nucleotide polymorphism that affects Drosha processing. AB - MicroRNAs (miRNAs) are a class of approximately 22-nucleotide noncoding RNAs that inhibit the expression of specific target genes at the posttranscriptional level. Recently, 11 miRNAs encoded by the pathogenic human herpesvirus Kaposi's sarcoma associated herpesvirus (KSHV) were cloned from latently infected cells. While the expression of these miRNAs has been confirmed by Northern analysis, their ability to inhibit target gene expression has not been demonstrated. We have devised a novel assay for miRNA function that uses lentiviral indicator vectors carrying two perfectly complementary target sites for each given miRNA in the 3' untranslated region of the Renilla luciferase gene. This assay allowed us to demonstrate the activity of each viral miRNA upon cotransduction of cells with the Renilla luciferase indicator vector together with a firefly luciferase control vector. In KSHV-infected BC-1 and BCBL-1 cells, but not uninfected control cells, Renilla luciferase expression was selectively reduced up to 10 fold. Interestingly, one of the viral miRNAs (miR-K5) exhibited much higher activity in BC-1 cells than in BCBL-1 cells. Sequence analysis of both viral genomes revealed a single nucleotide polymorphism in the miR-K5 precursor stem loop, which inhibits the expression of mature miR-K5 in BCBL-1 cells. We show that the primary miR-K5 sequence present in BCBL-1 results in diminished processing by Drosha both in vivo and in vitro. This is the first report of a naturally occurring sequence polymorphism in an miRNA precursor that results in reduced processing and therefore lower levels of mature miRNA expression and function. PMID- 16699013 TI - Evidence for functional protein interactions required for poliovirus RNA replication. AB - Poliovirus protein 2C contains a predicted N-terminal amphipathic helix that mediates association of the protein with the membranes of the viral RNA replication complex. A chimeric virus that contains sequences encoding the 18 residue core from the orthologous amphipathic helix from human rhinovirus type 14 (HRV14) was constructed. The chimeric virus exhibited defects in viral RNA replication and produced minute plaques on HeLa cell monolayers. Large plaque variants that contained mutations within the 2C-encoding region were generated upon subsequent passage. However, the majority of viruses that emerged with improved growth properties contained no changes in the region encoding 2C. Sequence analysis and reconstruction of genomes with individual mutations revealed changes in 3A or 2B sequences that compensated for the HRV14 amphipathic helix in the polio 2C-containing proteins, implying functional interactions among these proteins during the replication process. Direct binding between these viral proteins was confirmed by mammalian cell two-hybrid analysis. PMID- 16699014 TI - Gamma interferon plays a crucial early antiviral role in protection against West Nile virus infection. AB - West Nile virus (WNV) causes a severe central nervous system (CNS) infection in humans, primarily in the elderly and immunocompromised. Prior studies have established an essential protective role of several innate immune response elements, including alpha/beta interferon (IFN-alpha/beta), immunoglobulin M, gammadelta T cells, and complement against WNV infection. In this study, we demonstrate that a lack of IFN-gamma production or signaling results in increased vulnerability to lethal WNV infection by a subcutaneous route in mice, with a rise in mortality from 30% (wild-type mice) to 90% (IFN-gamma(-/-) or IFN-gammaR( /-) mice) and a decrease in the average survival time. This survival pattern in IFN-gamma(-/-) and IFN-gammaR(-/-) mice correlated with higher viremia and greater viral replication in lymphoid tissues. The increase in peripheral infection led to early CNS seeding since infectious WNV was detected several days earlier in the brains and spinal cords of IFN-gamma(-/-) or IFN-gammaR(-/-) mice. Bone marrow reconstitution experiments showed that gammadelta T cells require IFN gamma to limit dissemination by WNV. Moreover, treatment of primary dendritic cells with IFN-gamma reduced WNV production by 130-fold. Collectively, our experiments suggest that the dominant protective role of IFN-gamma against WNV is antiviral in nature, occurs in peripheral lymphoid tissues, and prevents viral dissemination to the CNS. PMID- 16699015 TI - Recombinant adenoviral vectors can induce expression of p73 via the E4-orf6/7 protein. AB - Despite the utility of recombinant adenoviral vectors in basic research, their therapeutic promise remains unfulfilled. Most engineered adenoviral vectors use a heterologous promoter to transcribe a foreign gene. We show that adenoviruses containing the cytomegalovirus immediate-early promoter induce the expression of the proapoptotic cellular protein TAp73 via the cyclin-dependent kinase retinoblastoma protein-E2F pathway in murine embryonic fibroblasts. Cells transduced with these vectors also expressed high levels of the adenoviral E4 orf6/7 and E2A proteins. By contrast, adenoviruses containing the ubiquitin C promoter failed to elicit these effects. E4-orf6/7 is necessary and sufficient for increased TAp73 expression, as shown by using retrovirus-mediated E4-orf6/7 expression and adenovirus with the E4-orf6/7 gene deleted. Activation of TAp73 likely occurs via E4-orf6/7-induced dimerization of E2F and subsequent binding to the inverted E2F-responsive elements within the TAp73 promoter. In addition, adenoviral vectors containing the cytomegalovirus immediate-early promoter, but not the ubiquitin C promoter, cooperated with chemotherapeutic agents to decrease cellularity in vitro. In contrast to murine embryonic fibroblasts, adenoviruses containing the ubiquitin C promoter, but not the cytomegalovirus immediate-early promoter, induced both E4-orf6/7 and TAp73 in human foreskin fibroblasts, emphasizing the importance of cellular context for promoter-dependent effects. Because TAp73 is important for the efficacy of chemotherapy, adenoviruses that increase TAp73 expression may enhance cancer therapies by promoting apoptosis. However, such adenoviruses may impair the long-term survival of transduced cells during gene replacement therapies. Our findings reveal previously unknown effects of foreign promoters in recombinant adenoviral vectors and suggest means to improve the utility of engineered adenoviruses by better controlling their impact on viral and cellular gene expression. PMID- 16699016 TI - Characterization of a permissive epitope insertion site in adenovirus hexon. AB - A robust immune response is generated against components of the adenovirus capsid. In particular, a potent and long-lived humoral response is elicited against the hexon protein. This is due to the efficient presentation of adenovirus capsid proteins to CD4+ T cells by antigen-presenting cells, in addition to the highly repetitive structure of the adenovirus capsids, which can efficiently stimulate B-cell proliferation. In the present study, we take advantage of this immune response by inserting epitopes against which an antibody response is desired into the adenovirus hexon. We use a B-cell epitope from Bacillus anthracis protective antigen (PA) as a model antigen to characterize hypervariable region 5 (HVR5) of hexon as a site for peptide insertion. We demonstrate that HVR5 can accommodate a peptide of up to 36 amino acids without adversely affecting virus infectivity, growth, or stability. Viruses containing chimeric hexons elicited antibodies against PA in mice, with total immunoglobulin G (IgG) titers reaching approximately 1 x 10(3) after two injections. The antibody response contained both IgG1 and IgG2a subtypes, suggesting that Th1 and Th2 immunity had been stimulated. Coinjection of wild-type adenovirus and a synthetic peptide from PA produced no detectable antibodies, indicating that incorporation of the epitope into the capsid was crucial for immune stimulation. Together, these results indicate that the adenovirus capsid is an efficient vehicle for presenting B-cell epitopes to the immune system, making this a useful approach for the design of epitope-based vaccines. PMID- 16699017 TI - Modulation of Kaposi's sarcoma-associated herpesvirus infection and replication by MEK/ERK, JNK, and p38 multiple mitogen-activated protein kinase pathways during primary infection. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) is etiologically associated with Kaposi's sarcoma, a dominant AIDS-related tumor of endothelial cells, and several other lymphoproliferative malignancies. While activation of the phosphatidylinositol 3-kinase-protein kinase C-MEK-ERK pathway is essential for KSHV infection, we have recently shown that KSHV also activates JNK and p38 mitogen-activated protein kinase (MAPK) pathways during primary infection (J. Xie, H. Y. Pan, S. Yoo, and S.-J. Gao, J. Virol. 79:15027-15037, 2005). Here, we found that activation of both JNK and p38 pathways was also essential for KSHV infection. Inhibitors of all three MAPK pathways reduced KSHV infectivity in both human umbilical vein endothelial cells (HUVEC) and 293 cells. These inhibitory effects were dose dependent and occurred at the virus entry stage of infection. Consistently, inhibition of all three MAPK pathways with dominant-negative constructs reduced KSHV infectivity whereas activation of the ERK pathway but not the JNK and p38 pathways enhanced KSHV infectivity. Importantly, inhibition of all three MAPK pathways also reduced the yield of infectious virions during KSHV productive infection of HUVEC. While the reduction of infectious virions was in part due to the reduced infectivity, it was also the result of direct modulation of KSHV lytic replication by the MAPK pathways. Accordingly, KSHV upregulated the expression of RTA (Orf50), a master transactivator of KSHV lytic replication, and activated its promoter during primary infection. Furthermore, KSHV activation of RTA promoter during primary infection was modulated by all three MAPK pathways, predominantly through their downstream target AP-1. Together, these results indicate that, by modulating multiple MAPK pathways, KSHV manipulates the host cells to facilitate its entry into the cells and postentry productive lytic replication during primary infection. PMID- 16699018 TI - Effect of apolipoprotein E on the cerebral load of latent herpes simplex virus type 1 DNA. AB - Herpes simplex virus type 1 (HSV-1) is neurotropic and enters a latent state lasting the lifetime of the host. This pathogen has recently been proposed as a risk factor for Alzheimer's disease (AD) in conjunction with apolipoprotein E4 (ApoE4). In a murine acute infection model, we showed that viral neuroinvasiveness depends directly on the overall ApoE dosage and especially on the presence of isoform ApoE4. If an interaction between ApoE and HSV-1 is involved in AD, it may occur during latency rather than during acute infection. Certainly, ApoE plays an important role in late-onset AD, i.e., at a time in life when the majority of people harbor HSV-1 in their nervous system. In the present work, wild-type, APOE knockout, APOE3, and APOE4 transgenic mice were used to analyze the influence of the ApoE profile on the levels of latent virus DNA. The knockout mice had significantly lower concentrations of the virus in the nervous system than the wild-type mice, while the APOE4 mice had very high levels in the brain compared to the APOE3 animals. ApoE4 seems to facilitate HSV-1 latency in the brain much more so than ApoE3. The APOE dosage correlated directly with the HSV-1 DNA concentration in the brain, strengthening the hypothesis that HSV-1, together with ApoE, might be involved in AD. PMID- 16699019 TI - Roles of human parainfluenza virus type 3 bases 13 to 78 in replication and transcription: identification of an additional replication promoter element and evidence for internal transcription initiation. AB - The genomic promoter of human parainfluenza virus type 3 (HPIV3) contains multiple cis-elements controlling transcription and replication. Previous work showed that regions 1 to 12 and 79 to 96 were critical in promoting replication of an HPIV3 minireplicon, while the intergenic sequence and N gene start signal (IS/Ngs, bases 49 to 61) were important for transcription. Because these data were collected primarily using point mutations, not every base from position 1 to 96 was analyzed, and some important control elements may have been missed. To clarify the role of bases 13 to 78 in transcription and replication, a series of mutations were made which collectively scanned this entire region. Mutation of bases 13 to 28 resulted in markedly decreased HPIV3 minireplicon replication, indicating these bases constitute an additional cis-element involved in the synthesis of the HPIV3 antigenomic RNA. The position dependence of the IS/Ngs was also examined. Analysis of mutants in which the IS/Ngs was shifted 5' or 3' showed that this segment could be moved without significantly disrupting transcription initiation. Additional mutants which contained two successive IS/Ngs segments were created to test whether the polymerase accessed the gene start signal by proceeding along the template 3' to 5' or by binding internally at the gene start signal. Based on analysis of the double gene start mutants, we propose a model of internal transcription initiation in which the polymerase enters the template at approximately the location of the natural N gene start but then scans the template bidirectionally to find a gene start signal and initiate transcription. PMID- 16699020 TI - A short isoform of human cytomegalovirus US3 functions as a dominant negative inhibitor of the full-length form. AB - Human cytomegalovirus encodes four unique short (US) region proteins, each of which is independently sufficient for causing the down-regulation of major histocompatibility complex (MHC) class I molecules on the cell surface. This down regulation enables infected cells to evade recognition by cytotoxic T lymphocytes (CTLs) but makes them vulnerable to lysis by natural killer (NK) cells, which lyse those cells that lack MHC class I molecules. The 22-kDa US3 glycoprotein is able to down-regulate the surface expression of MHC class I molecules by dual mechanisms: direct endoplasmic reticulum retention by physical association and/or tapasin inhibition. The alternative splicing of the US3 gene generates two additional products, including 17-kDa and 3.5-kDa truncated isoforms; however, the functional significance of these isoforms during viral infection is unknown. Here, we describe a novel mode of self-regulation of US3 function that uses the endogenously produced truncated isoform. The truncated isoform itself neither binds to MHC class I molecules nor prevents the full-length US3 from interacting with MHC class I molecules. Instead, the truncated isoform associates with tapasin and competes with full-length US3 for binding to tapasin; thus, it suppresses the action of US3 that causes the disruption of the function of tapasin. Our results indicate that the truncated isoform of the US3 locus acts as a dominant negative regulator of full-length US3 activity. These data reflect the manner in which the virus has developed temporal survival strategies during viral infection against immune surveillance involving both CTLs and NK cells. PMID- 16699021 TI - Inhibition of DNA replication of human papillomavirus by artificial zinc finger proteins. AB - Recently, we demonstrated that plant DNA virus replication was inhibited in planta by using an artificial zinc finger protein (AZP) and created AZP-based transgenic plants resistant to DNA virus infection. Here we apply the AZP technology to the inhibition of replication of a mammalian DNA virus, human papillomavirus type 18 (HPV-18). Two AZPs, designated AZP(HPV)-1 and AZP(HPV)-2, were designed by using our nondegenerate recognition code table and were constructed to block binding of the HPV-18 E2 replication protein to the replication origin. Both of the newly designed AZPs had much higher affinities towards the replication origin than did the E2 protein, and they efficiently blocked E2 binding in vitro. In transient replication assays, both AZPs inhibited viral DNA replication, especially AZP(HPV)-2, which reduced the replication level to approximately 10%. We also demonstrated in transient replication assays, using plasmids with mutant replication origins, that AZP(HPV)-2 could precisely recognize the replication origin in mammalian cells. Thus, it was demonstrated that the AZP technology could be applied not only to plant DNA viruses but also to mammalian DNA viruses. PMID- 16699022 TI - Molecular determinants of substrate specificity for Semliki Forest virus nonstructural protease. AB - The C-terminal cysteine protease domain of Semliki Forest virus nonstructural protein 2 (nsP2) regulates the virus life cycle by sequentially cleaving at three specific sites within the virus-encoded replicase polyprotein P1234. The site between nsP3 and nsP4 (the 3/4 site) is cleaved most efficiently. Analysis of Semliki Forest virus-specific cleavage sites with shuffled N-terminal and C terminal half-sites showed that the main determinants of cleavage efficiency are located in the region preceding the cleavage site. Random mutagenesis analysis revealed that amino acid residues in positions P4, P3, P2, and P1 of the 3/4 cleavage site cannot tolerate much variation, whereas in the P5 position most residues were permitted. When mutations affecting cleavage efficiency were introduced into the 2/3 and 3/4 cleavage sites, the resulting viruses remained viable but had similar defects in P1234 processing as observed in the in vitro assay. Complete blockage of the 3/4 cleavage was found to be lethal. The amino acid in position P1' had a significant effect on cleavage efficiency, and in this regard the protease markedly preferred a glycine residue over the tyrosine natively present in the 3/4 site. Therefore, the cleavage sites represent a compromise between protease recognition and other requirements of the virus life cycle. The protease recognizes at least residues P4 to P1', and the P4 arginine residue plays an important role in the fast cleavage of the 3/4 site. PMID- 16699023 TI - Deletion of open reading frame UL26 from the human cytomegalovirus genome results in reduced viral growth, which involves impaired stability of viral particles. AB - We previously showed that open reading frame (ORF) UL26 of human cytomegalovirus, a member of the US22 multigene family of betaherpesviruses, encodes a novel tegument protein, which is imported into cells in the course of viral infection. Moreover, we demonstrated that pUL26 contains a strong transcriptional activation domain and is capable of stimulating the major immediate-early (IE) enhancer promoter. Since this suggested an important function of pUL26 during the initiation of the viral replicative cycle, we sought to ascertain the relevance of pUL26 by construction of a viral deletion mutant lacking the UL26 ORF using the bacterial artificial chromosome mutagenesis procedure. The resulting deletion virus was verified by PCR, enzyme restriction, and Southern blot analyses. After infection of human foreskin fibroblasts, the UL26 deletion mutant showed a small plaque phenotype and replicated to significantly lower titers than wild-type or revertant virus. In particular, we noticed a striking decrease of infectious titers 7 days postinfection in a multistep growth experiment, whereas the release of viral DNA from infected cells was not impaired. A further investigation of this aspect revealed a significantly diminished stability of viral particles derived from the UL26 deletion mutant. Consistent with this, we observed that the tegument composition of the deletion mutant deviates from that of the wild-type virus. We therefore hypothesize that pUL26 plays a role not only in the onset of IE gene transcription but also in the assembly of the viral tegument layer in a stable and correct manner. PMID- 16699024 TI - High-efficiency system for the construction of adenovirus vectors and its application to the generation of representative adenovirus-based cDNA expression libraries. AB - We here describe a convenient system for the production of recombinant adenovirus vectors and its use for the construction of a representative adenovirus-based cDNA expression library. The system is based on direct site-specific insertion of transgene cassettes into a replicating donor virus. The transgene is inserted into a donor plasmid containing the viral 5' inverted terminal repeat, the complete viral packaging signal, and a single loxP site. The plasmid is then transfected into a Cre recombinase-expressing packaging cell line that has been infected with a donor virus containing a partially deleted packaging signal flanked by loxP sites. Cre recombinase, by two steps of action, sequentially catalyzes the generation of a nonpackageable donor virus acceptor substrate and the generation of the desired recombinant adenovirus vector. Due to its growth impairment, residual donor virus can efficiently be counterselected during amplification of the recombinant adenovirus vector. By using this adenovirus construction system, a plasmid-based human liver cDNA library was converted by a single step into an adenovirus-based cDNA expression library with about 10(6) independent adenovirus clones. The high-titer purified library was shown to contain about 44% of full-length cDNAs with an average insert size of 1.3 kb. cDNAs of a gene expressed at a high level (human alpha(1)-antitrypsin) and a gene expressed at a relatively low level (human coagulation factor IX) in human liver were isolated from the adenovirus-based library using an enzyme-linked immunosorbent assay-based screening procedure. PMID- 16699025 TI - Nuclear localization of flavivirus RNA synthesis in infected cells. AB - Flaviviral replication is believed to be exclusively cytoplasmic, occurring within virus-induced membrane-bound replication complexes in the host cytoplasm. Here we show that a significant proportion (20%) of the total RNA-dependent RNA polymerase (RdRp) activity from cells infected with West Nile virus, Japanese encephalitis virus (JEV), and dengue virus is resident within the nucleus. Consistent with this, the major replicase proteins NS3 and NS5 of JEV also localized within the nucleus. NS5 was found distributed throughout the nucleoplasm, but NS3 was present at sites of active flaviviral RNA synthesis, colocalizing with NS5, and visible as distinct foci along the inner periphery of the nucleus by confocal and immunoelectron microscopy. Both these viral replicase proteins were also present in the nuclear matrix, colocalizing with the peripheral lamina, and revealed a well-entrenched nuclear location for the viral replication complex. In keeping with this observation, antibodies to either NS3 or NS5 coimmunoprecipitated the other protein from isolated nuclei along with newly synthesized viral RNA. Taken together these data suggest an absolute requirement for both of the replicase proteins for nucleus-localized synthesis of flavivirus RNA. Thus, we conclusively demonstrate for the first time that the host cell nucleus functions as an additional site for the presence of functionally active flaviviral replicase complex. PMID- 16699026 TI - Lipid-mediated introduction of hepatitis B virus capsids into nonsusceptible cells allows highly efficient replication and facilitates the study of early infection events. AB - The hepatitis B virus (HBV) is an enveloped DNA virus which is highly infectious in vivo. In vitro, only primary hepatocytes of humans and Tupaia belangeri or the novel HepaRG cell line are susceptible to HBV, but infection is inefficient and study of early infection events in single cells is unsatisfactory. Since hepatoma cells replicate the virus efficiently after transfection, this limited infection efficiency must be related to the initial entry phase. Here, we describe the lipid-based delivery of HBV capsids into nonsusceptible cells, circumventing the natural entry pathway. Successful infection was monitored by observing the emergence of the nuclear viral covalently closed circular DNA and the production of progeny virus and subviral particles. Lipid-mediated transfer initiated productive infection that was at least 100-fold more effective than infection of permissive cell cultures. High-dose capsid transfer showed that the uptake was not receptor limited and allowed the intracellular transport of capsids and genomes to be examined microscopically. The addition of inhibitors confirmed an entry pathway by fusion of the lipid with the plasma membrane. By indirect immune fluorescence and native fluorescence in situ hybridization, we followed the pathway of capsids and viral genomes in individual cells. We observed an active microtubule-dependent capsid transfer to the nucleus and a subsequent release of the viral genomes exclusively into the karyoplasm. Lipid-mediated transfer of viral capsids thus appears to allow efficient introduction of genetic information into target cells, facilitating studies of early infection events which are otherwise impeded by the small number of viruses entering the cell. PMID- 16699027 TI - Lepidopteran ortholog of Drosophila breathless is a receptor for the baculovirus fibroblast growth factor. AB - The Bombyx mori nucleopolyhedrovirus (BmNPV) encodes a gene homologous to the mammalian fibroblast growth factor (FGF) family. We report the cloning of B. mori and Spodoptera frugiperda orthologous genes (Bmbtl and Sfbtl, respectively) of Drosophila melanogaster breathless (btl) encoding a receptor for Branchless/FGF and show that these genes encode the receptor for a baculovirus-encoded FGF (vFGF). Sequence analysis showed that BmBtl is composed of 856 amino acid residues, which potentially encodes a 97.3-kDa polypeptide and shares structural features and sequence similarities with the FGF receptor family. Reverse transcription-PCR experiments showed that Bmbtl was abundantly expressed in the trachea and midgut in B. mori larvae, with moderate expression observed in the hemocytes and the B. mori cultured cell line BmN. We generated Sf-9 cells that stably expressed His-tagged BmBtl. Western blot analysis revealed that BmBtl was an approximately 110-kDa protein. Immunoprecipitation experiments showed that BmNPV vFGF markedly phosphorylated BmBtl in Sf-9 cells. In addition, we found that BmBtl overexpression enhanced the migration activity for BmNPV vFGF. Furthermore, we generated Sf-9 cells in which Sfbtl was knocked down by transfection with double-strand RNA-expressing plasmids. In these cells, cell motility triggered by vFGF was markedly reduced. These results strongly suggest that the Btl orthologs, BmBtl and SfBtl, are the receptors for vFGF, which mediate vFGF-induced host cell chemotaxis. PMID- 16699028 TI - Expression profiles of bovine adeno-associated virus and avian adeno-associated virus display significant similarity to that of adeno-associated virus type 5. AB - We present the first detailed expression profiles of nonprimate-derived adeno associated viruses, namely, bovine adeno-associated virus (B-AAV) and avian adeno associated virus (A-AAV), which were obtained after the infection of cell lines derived from their natural hosts. In general, the profiles of B-AAV and A-AAV were quite similar to that of AAV5; however, both exhibited features found for AAV2 as well. Like adeno-associated virus type 5 (AAV5), B-AAV and A-AAV utilized an internal polyadenylation site [(pA)p]; however, it was used to greater relative levels by B-AAV than by A-AAV. Similar to AAV5, >99% of B-AAV RNAs generated from upstream promoters were polyadenylated at (pA)p and hence not spliced. In contrast, ca. 50% of the A-AAV RNAs generated from upstream promoters read through (pA)p, as seen for AAV2. However, A-AAV generated lower levels of spliced P5 and P19 products than does AAV2, suggesting that A-AAV generates lower relative levels of Rep 68 and Rep 40. An additional difference in the expression profile of these viruses was that B-AAV generated a greater level of ITR initiated RNAs than did A-AAV or AAV5. In addition, we demonstrate that, like AAV2, transactivation of transcription of the capsid-gene promoter of B-AAV required both adenovirus and targeting of its Rep protein to the transcription template; however, expression of the capsid-gene promoter of A-AAV was, like AAV5, largely independent of both adenovirus and its Rep proteins. PMID- 16699029 TI - The Herpesvirus capsid surface protein, VP26, and the majority of the tegument proteins are dispensable for capsid transport toward the nucleus. AB - Upon entering a cell, alphaherpesvirus capsids are transported toward the minus ends of microtubules and ultimately deposit virus DNA within the host nucleus. The virus proteins that mediate this centripetal transport are unknown but are expected to be either viral tegument proteins, which are a group of capsid associated proteins, or a surface component of the capsid itself. Starting with derivatives of pseudorabies virus that encode a fluorescent protein fused to a structural component of the virus, we have made a collection of 12 mutant viruses that lack either the VP26 capsid protein or an individual tegument protein. Using live-cell fluorescence microscopy, we tracked individual virus particles in axons following infection of primary sensory neurons. Quantitative analysis of the VP26 null virus indicates that this protein plays no observable role in capsid transport. Furthermore, viruses lacking tegument proteins that are nonessential for virus propagation in cell culture were also competent for axonal transport. These results indicate that a protein essential for viral propagation mediates transport of the capsid to the nucleus. PMID- 16699030 TI - Prediction and identification of herpes simplex virus 1-encoded microRNAs. AB - MicroRNAs (miRNAs) are key regulators of gene expression in higher eukaryotes. Recently, miRNAs have been identified from viruses with double-stranded DNA genomes. To attempt to identify miRNAs encoded by herpes simplex virus 1 (HSV-1), we applied a computational method to screen the complete genome of HSV-1 for sequences that adopt an extended stem-loop structure and display a pattern of nucleotide divergence characteristic of known miRNAs. Using this method, we identified 11 HSV-1 genomic loci predicted to encode 13 miRNA precursors and 24 miRNA candidates. Eight of the HSV-1 miRNA candidates were predicted to be conserved in HSV-2. The precursor and the mature form of one HSV-1 miRNA candidate, which is encoded approximately 450 bp upstream of the transcription start site of the latency-associated transcript (LAT), were detected during infection of Vero cells by Northern blot hybridization. These RNAs, which behave as late gene products, are not predicted to be conserved in HSV-2. Additionally, small RNAs, including some that are roughly the expected size of precursor miRNAs, were detected using probes for miRNA candidates derived from sequences encoding the 8.3-kilobase LAT, from sequences complementary to U(L)15 mRNA, and from the region between ICP4 and U(S)1. However, no species the size of typical mature miRNAs were detected using these probes. Three of these latter miRNA candidates were predicted to be conserved in HSV-2. Thus, HSV-1 encodes at least one miRNA. We hypothesize that HSV-1 miRNAs regulate viral and host gene expression. PMID- 16699032 TI - Gangliosides are essential for bovine adeno-associated virus entry. AB - Recombinant adeno-associated viruses (AAV) are promising gene therapy vectors. We have recently identified a bovine adeno-associated virus (BAAV) that demonstrates unique tropism and transduction activity compared to primate AAVs. To better understand the entry pathway and cell tropism of BAAV, we have characterized the initial cell surface interactions required for transduction with BAAV vectors. Like a number of AAVs, BAAV requires cell surface sialic acid groups for transduction and virus attachment. However, glycosphingolipids (GSLs), not cell surface proteins, were required for vector entry and transduction. Incorporation of gangliosides, ceramide-based glycolipids containing one or more sialic acid groups, into the cytoplasmic cell membranes of GSL-depleted COS cells partially reconstituted BAAV transduction. The dependency of BAAV on gangliosides for transduction was further confirmed by studies with C6 cells, a rat glioma cell line that is deficient in the synthesis of complex gangliosides. C6 cells were resistant to transduction by BAAV. Addition of gangliosides to C6 cells prior to transduction rendered the cells susceptible to transduction by BAAV. Therefore, gangliosides are a likely receptor for BAAV. PMID- 16699031 TI - Diversity of the CD8+ T-cell response to herpes simplex virus type 2 proteins among persons with genital herpes. AB - Cytolytic T cells play a major role in controlling herpes simplex virus type 2 (HSV-2) infections in humans. In an effort to more thoroughly evaluate the response to HSV-2 directly, ex vivo, we developed an enzyme-linked immunospot (ELISPOT) assay that utilized pools of overlapping synthetic peptides presented by autologous dendritic cells to purified CD8(+) T cells. Donor response rates to individual open reading frames (ORFs) ranged from fewer than 5% responding to as many as 70% responding, with the greatest frequency of responses (by ORF) being directed against UL39, UL25, UL27, ICP0, UL46, and UL47 in descending order of frequency. HSV-2-seropositive subjects responded to as few as 3 or as many as 46 of the 48 ORFs tested, with a median of 11 ORFs recognized. HLA-B*07 expression correlated with stronger responses overall that were directed primarily against UL49 and UL46. Cumulative precursor frequencies in the blood ranged from 500 to almost 6,000 HSV-2 spot-forming units/10(6) CD8(+) T cells. The magnitude and breadth of the response in the infected population were greater than previously appreciated. Whether this variability in the CD8(+) T-cell response within individuals is associated with the frequency of viral reactivation warrants further study. PMID- 16699033 TI - Epitopes expressed in different adenovirus capsid proteins induce different levels of epitope-specific immunity. AB - On the basis of the concept that the capsid proteins of adenovirus (Ad) gene transfer vectors can be genetically manipulated to enhance the immunogenicity of Ad-based vaccines, the present study compared the antiantigen immunogenicity of Ad vectors with a common epitope of the hemagglutinin (HA) protein of the influenza A virus incorporated into the outer Ad capsid protein hexon, penton base, fiber knob, or protein IX. Incorporation of the same epitope into the different capsid proteins provided insights into the correlation between epitope position and antiepitope immunity. Following immunization of three different strains of mice (C57BL/6, BALB/c, and CBA) with either an equal number of Ad particles (resulting in a different total HA copy number) or different Ad particle numbers (to achieve the same HA copy number), the highest primary (immunoglobulin M [IgM]) and secondary (IgG) anti-HA humoral and cellular CD4 gamma interferon and interleukin-4 responses against HA were always achieved with the Ad vector carrying the HA epitope in fiber knob. These observations suggest that the immune response against an epitope inserted into Ad capsid proteins is not necessarily dependent on the capsid protein number and imply that the choice of incorporation site in Ad capsid proteins in their use as vaccines needs to be compared in vivo. PMID- 16699034 TI - A herpes simplex virus recombinant that exhibits a single-chain antibody to HER2/neu enters cells through the mammary tumor receptor, independently of the gD receptors. AB - The human epidermal growth factor receptor 2/neuregulin (HER2/neu) receptor is overexpressed in highly malignant mammary and ovarian tumors and correlates with a poor prognosis. It is a target for therapy; humanized monoclonal antibodies to HER2 have led to increased survival of patients with HER2/neu-positive breast cancer. As a first step in the design of an oncolytic herpes simplex virus able to selectively infect HER2/neu-positive cells, we constructed two recombinants, R LM11 and R-LM11L, that carry a single-chain antibody (scFv) against HER2 inserted at residue 24 of gD. The inserts were 247 or 256 amino acids long, and the size of the gD ectodomain was almost doubled by the insertion. We report the following. R-LM11 and R-LM11L infected derivatives of receptor-negative J or CHO cells that expressed HER2/neu as the sole receptor. Entry was dependent on HER2/neu, since it was inhibited in a dose-dependent manner by monoclonal antibodies to HER2/neu and by a soluble form of the receptor. The scFv insertion in gD disrupted the ability of the virus to enter cells through HVEM but maintained the ability to enter through nectin1. This report provides proof of principle that gD can tolerate fusion to a heterologous protein almost as large as the gD ectodomain itself without loss of profusion activity. Because the number of scFv's to a variety of receptors is continually increasing, this report makes possible the specific targeting of herpes simplex virus to a large collection of cell surface molecules for both oncolytic activity and visualization of tumor cells. PMID- 16699035 TI - Furin cleavage potentiates the membrane fusion-controlling intersubunit disulfide bond isomerization activity of leukemia virus Env. AB - The membrane fusion protein of murine leukemia virus is a trimer of a disulfide linked peripheral-transmembrane (SU-TM) subunit complex. The intersubunit disulfide bond is in SU linked to a disulfide bond isomerization motif, CXXC, with which the virus controls its fusion reaction (M. Wallin, M. Ekstrom, and H. Garoff, EMBO J. 23:54-65, 2004). Upon receptor binding the isomerase rearranges the intersubunit disulfide bond into a disulfide bond isomer within the motif. This facilitates SU dissociation and fusion activation in the TM subunit. In the present study we have asked whether furin cleavage of the Env precursor potentiates the isomerase to be triggered. To this end we accumulated the late form of the precursor, gp90, in the cell by incubation in the presence of a furin inhibiting peptide. The isomerization was done by NP-40 incubation or by a heat pulse under alkylation-free conditions. The cells were lysed in the presence of alkylator, and the precursor was immunoprecipitated, gel isolated, deglycosylated, and subjected to complete trypsin digestion. Disulfide-linked peptide complexes were separated by sodium dodecyl sulfate-tricine-polyacrylamide gel electrophoresis under nonreducing conditions. This assay revealed the size of the characteristic major disulfide-linked peptide complex that differentiates the two isomers of the disulfide bond between Cys336 (or Cys339) and Cys563, i.e., the bond corresponding to the intersubunit disulfide bond. The analyses showed that the isomerase was five- to eightfold more resistant to triggering in the precursor than in the mature, cleaved form. This suggests that the isomerase becomes potentiated for triggering by a structural change in Env that is induced by furin cleavage in the cell. PMID- 16699036 TI - Sequential immunization with V3 peptides from primary human immunodeficiency virus type 1 produces cross-neutralizing antibodies against primary isolates with a matching narrow-neutralization sequence motif. AB - An antibody response capable of neutralizing not only homologous but also heterologous forms of the CXCR4-tropic human immunodeficiency virus type 1 (HIV 1) MNp and CCR5-tropic primary isolate HIV-1 JR-CSF was achieved through sequential immunization with a combination of synthetic peptides representing HIV 1 Env V3 sequences from field and laboratory HIV-1 clade B isolates. In contrast, repeated immunization with a single V3 peptide generated antibodies that neutralized only type-specific laboratory-adapted homologous viruses. To determine whether the cross-neutralization response could be attributed to a cross-reactive antibody in the immunized animals, we isolated a monoclonal antibody, C25, which neutralized the heterologous primary viruses of HIV-1 clade B. Furthermore, we generated a humanized monoclonal antibody, KD-247, by transferring the genes of the complementary determining region of C25 into genes of the human V region of the antibody. KD-247 bound with high affinity to the "PGR" motif within the HIV-1 Env V3 tip region, and, among the established reference antibodies, it most effectively neutralized primary HIV-1 field isolates possessing the matching neutralization sequence motif, suggesting its promise for clinical applications involving passive immunizations. These results demonstrate that sequential immunization with B-cell epitope peptides may contribute to a humoral immune-based HIV vaccine strategy. Indeed, they help lay the groundwork for the development of HIV-1 vaccine strategies that use sequential immunization with biologically relevant peptides to overcome difficulties associated with otherwise poorly immunogenic epitopes. PMID- 16699038 TI - Influence of pseudorabies virus proteins on neuroinvasion and neurovirulence in mice. AB - Neurotropism is a distinctive feature of members of the Alphaherpesvirinae. However, its molecular basis remains enigmatic. In the past, research has been focused mainly on the role of viral envelope proteins in modulating herpesvirus neuroinvasion and neurovirulence (T. C. Mettenleiter, Virus Res. 92:192-206, 2003). To further analyze the molecular requirements for neuroinvasion of the alphaherpesvirus pseudorabies virus (PrV), adult mice were infected intranasally with a set of single- or multiple-deletion mutants lacking the UL3, UL4, UL7, UL11, UL13, UL16, UL17, UL21, UL31, UL34, UL37, UL41, UL43, UL46, UL47, UL48, UL51, US3, US9, glycoprotein E (gE), gM, UL11/US9, UL11/UL16, UL16/UL21, UL11/UL16/UL21, UL11/gE, UL11/gM, UL43/gK, UL43/gM, or UL43/gK/gM genes. Neurovirulence was evaluated by measuring mean survival times compared to that after wild-type virus infection. Furthermore, by immunohistochemical detection of infected neurons, the kinetics of viral spread in the murine central nervous system was investigated. PMID- 16699037 TI - Anti-V3 humanized antibody KD-247 effectively suppresses ex vivo generation of human immunodeficiency virus type 1 and affords sterile protection of monkeys against a heterologous simian/human immunodeficiency virus infection. AB - In an accompanying report (Y. Eda, M. Takizawa, T. Murakami, H. Maeda, K. Kimachi, H. Yonemura, S. Koyanagi, K. Shiosaki, H. Higuchi, K. Makizumi, T. Nakashima, K. Osatomi, S. Tokiyoshi, S. Matsushita, N. Yamamoto, and M. Honda, J. Virol. 80:5552-5562, 2006), we discuss our production of a high-affinity humanized monoclonal antibody, KD-247, by sequential immunization with V3 peptides derived from human immunodeficiency virus type 1 (HIV-1) clade B primary isolates. Epitope mapping revealed that KD-247 recognized the Pro-Gly-Arg V3 tip sequence conserved in HIV-1 clade B isolates. In this study, we further demonstrate that in vitro, KD-247 efficiently neutralizes CXCR4- and CCR5-tropic primary HIV-1 clade B and clade B' with matching neutralization sequence motifs but does not neutralize sequence-mismatched clade B and clade E isolates. Monkeys were provided sterile protection against heterologous simian/human immunodeficiency virus challenge by the passive transfer of a single high dose (45 mg per kg of body weight) of KD-247 and afforded partial protection by lower antibody doses (30 and 15 mg per kg). Protective neutralization endpoint titers in plasma at the time of virus challenge were 1:160 in animals passively transferred with a high dose of the antibody. The antiviral efficacy of the antibody was further confirmed by its suppression of the ex vivo generation of primary HIV-1 quasispecies in peripheral blood mononuclear cell cultures from HIV infected individuals. Therefore, KD-247 promises to be a valuable tool not only as a passive immunization antibody for the prevention of HIV infection but also as an immunotherapy for the suppression of HIV in phenotype-matched HIV-infected individuals. PMID- 16699039 TI - Varicella-zoster virus infection of human foreskin fibroblast cells results in atypical cyclin expression and cyclin-dependent kinase activity. AB - In its course of human infection, varicella-zoster virus (VZV) infects rarely dividing cells such as dermal fibroblasts, differentiated keratinocytes, mature T cells, and neurons, none of which are actively synthesizing DNA; however, VZV is able to productively infect them and use their machinery to replicate the viral genome. We hypothesized that VZV alters the intracellular environment to favor viral replication by dysregulating cell cycle proteins and kinases. Cyclin dependent kinases (CDKs) and cyclins displayed a highly unusual profile in VZV infected confluent fibroblasts: total amounts of CDK1, CDK2, cyclin B1, cyclin D3, and cyclin A protein increased, and kinase activities of CDK2, CDK4, and cyclin B1 were strongly and simultaneously induced. Cyclins B1 and D3 increased as early as 24 h after infection, concurrent with VZV protein synthesis. Confocal microscopy indicated that cyclin D3 overexpression was limited to areas of IE62 production, whereas cyclin B1 expression was irregular across the VZV plaque. Downstream substrates of CDKs, including pRb, p107, and GM130, did not show phosphorylation by immunoblotting, and p21 and p27 protein levels were increased following infection. Finally, although the complement of cyclin expression and high CDK activity indicated a progression through the S and G(2) phases of the cell cycle, DNA staining and flow cytometry indicated a possible G(1)/S blockade in infected cells. These data support earlier studies showing that pharmacological CDK inhibitors can inhibit VZV replication in cultured cells. PMID- 16699040 TI - Human cytomegalovirus attenuates interleukin-1beta and tumor necrosis factor alpha proinflammatory signaling by inhibition of NF-kappaB activation. AB - Viral infection is associated with a vigorous inflammatory response characterized by cellular infiltration and release of the proinflammatory cytokines interleukin 1 (IL-1) and tumor necrosis factor alpha (TNF-alpha). In the present study, we identified a novel function of human cytomegalovirus (HCMV) that results in inhibition of IL-1 and TNF-alpha signaling pathways. The effect on these pathways was limited to cells infected with the virus, occurred at late times of infection, and was independent of cell type or virus strain. IL-1 and TNF-alpha signaling pathways converge at a point upstream of NF-kappaB activation and involve phosphorylation and degradation of the NF-kappaB inhibitory molecule IkappaBalpha. The HCMV inhibition of IL-1 and TNF-alpha pathways corresponded to a suppression of NF-kappaB activation. Analysis of IkappaBalpha phosphorylation and degradation suggested that HCMV induced two independent blocks in NF-kappaB activation, which occurred upstream from the point of convergence of the IL-1 and TNF-alpha pathways. We believe that the ability of HCMV to inhibit these two major proinflammatory pathways reveals a critical aspect of HCMV biology, with possible importance for immune evasion, as well as establishment of infection in cell types persistently infected by this virus. PMID- 16699041 TI - Silencing of hepatitis A virus infection by small interfering RNAs. AB - Infection by hepatitis A virus (HAV) can cause acute hepatitis and, rarely, fulminant liver failure, in particular in patients chronically infected with hepatitis C virus. Based on our previous observation that small interfering RNAs (siRNAs) can silence translation and replication of the firefly luciferase encoding HAV replicon, we now exploited this technology to demonstrate the effect of siRNAs on viral infection in Huh-7 cells. Freshly and persistently infected cells were transfected with siRNAs targeting various sites in the HAV nonstructural genes. Compared to a single application, consecutive siRNA transfections targeting multiple sequences in the viral genome resulted in a more efficient and sustained silencing effect than a single transfection. In most instances, multiple applications of a single siRNA led to the emergence of viral escape mutants with mutated target sites that rendered these genomes resistant to RNA interference (RNAi). Efficient and sustained suppression of the viral infectivity was achieved after consecutive applications of an siRNA targeting a computer-predicted hairpin structure. This siRNA holds promise as a therapeutic tool for severe courses of HAV infection. In addition, the results provide new insight into the structural bases for sequence-specific RNAi. PMID- 16699042 TI - Sequence requirements for localization of human cytomegalovirus tegument protein pp28 to the virus assembly compartment and for assembly of infectious virus. AB - The human cytomegalovirus UL99 open reading frame encodes a 190-amino-acid (aa) tegument protein, pp28, that is myristoylated and phosphorylated. pp28 is essential for assembly of infectious virus, and nonenveloped virions accumulate in the cytoplasm of cells infected with recombinant viruses with a UL99 deletion. pp28 is localized to the endoplasmic reticulum-Golgi intermediate compartment (ERGIC) in transfected cells, while in infected cells, it is localized together with other virion proteins in a juxtanuclear compartment termed the assembly compartment (AC). We investigated the sequence requirements for pp28 trafficking to the AC and assembly of infectious virus. Our studies indicated that the first 30 to 35 aa were required for localization of pp28 to the ERGIC in transfected cells. Mutant forms of pp28 containing only the first 35 aa localized with other virion structural proteins to cytoplasmic compartments early in infection, but localization to the AC at late times required a minimum of 50 aa. In agreement with previous reports, we demonstrated that the deletion of a cluster of acidic amino acids (aa 44 to 59) prevented wild-type trafficking of pp28 and recovery of infectious virus. A recombinant virus expressing only the first 50 aa was replication competent, and this mutant, pp28, localized to the AC in cells infected with this virus. These findings argued that localization of pp28 to the AC was essential for assembly of infectious virus and raised the possibility that amino acids in the amino terminus of pp28 have additional roles in the envelopment and assembly of the virion other than simply localizing pp28 to the AC. PMID- 16699043 TI - Batai and Ngari viruses: M segment reassortment and association with severe febrile disease outbreaks in East Africa. AB - Ngari virus is an orthobunyavirus recently recognized as a reassortant between Bunyamwera virus and an as yet unidentified M segment donor. Analysis of M segment sequences of Batai and Ilesha viruses revealed 95% deduced amino acid identity between Batai virus and Ngari virus. These findings suggest Batai virus as the donor of Ngari virus M segment sequence. Analysis of Batai virus-related African isolates identified UgMP-6830, isolated from mosquitoes in Uganda, as an isolate of Batai virus. KV-141, isolated during a febrile disease outbreak in Sudan, was identified as another isolate of Ngari virus, emphasizing a role of this reassortant virus in severe human illness throughout East Africa. PMID- 16699044 TI - Two surface-exposed elements of the B30.2/SPRY domain as potency determinants of N-tropic murine leukemia virus restriction by human TRIM5alpha. AB - Human TRIM5alpha (TRIM5alpha(hu)) potently restricts N-tropic (N-MLV), but not B tropic, murine leukemia virus in a manner dependent upon residue 110 of the viral capsid. Rhesus monkey TRIM5alpha (TRIM5alpha(rh)) inhibits N-MLV only weakly. The study of human-monkey TRIM5alpha chimerae revealed that both the v1 and v3 variable regions of the B30.2/SPRY domain contain potency determinants for N-MLV restriction. These variable regions are predicted to be surface-exposed elements on one face of the B30.2 domain. Acidic residues in v3 complement basic residue 110 of the N-MLV capsid. The results support recognition of the retroviral capsid by the TRIM5alpha B30.2 domain. PMID- 16699045 TI - Group B coxsackievirus diabetogenic phenotype correlates with replication efficiency. AB - Group B coxsackieviruses can initiate rapid onset type 1 diabetes (T1D) in old nonobese diabetic (NOD) mice. Inoculating high doses of poorly pathogenic CVB3/GA per mouse initiated rapid onset T1D. Viral protein was detectable in islets shortly after inoculation in association with beta cells as well as other primary islet cell types. The virulent strain CVB3/28 replicated to higher titers more rapidly than CVB3/GA in the pancreas and in established beta cell cultures. Exchange of 5'-nontranslated regions between the two CVB3 strains demonstrated a variable impact on replication in beta cell cultures and suppression of in vivo replication for both strains. While any CVB strain may be able to induce T1D in prediabetic NOD mice, T1D onset is linked both to the viral replication rate and infectious dose. PMID- 16699046 TI - Measles virus V protein inhibits p53 family member p73. AB - Paramyxovirus V proteins function as host interference factors that inactivate antiviral responses, including interferon. Characterization of cellular proteins that copurify with ectopically expressed measles virus V protein has revealed interactions with DNA binding domains of p53 family proteins, p53 and p73. Specific transcriptional assays reveal that expression of measles virus V cDNA inhibits p73, but not p53. Expression of measles virus V cDNA can delay cell death induced by genotoxic stress and also can decrease the abundance of the proapoptotic factor PUMA, a p73 target. Recombinant measles virus with an engineered deficiency in V protein is capable of inducing more severe cytopathic effects than the wild type, implicating measles virus V protein as an inhibitor of cell death. These findings also suggest that p73-PUMA signaling may be a previously unrecognized arm of cellular innate antiviral immunity. PMID- 16699047 TI - Transspecies transmission of the endogenous koala retrovirus. AB - The koala retrovirus (KoRV) is a gammaretrovirus closely related to the gibbon ape leukemia virus and induces leukemias and immune deficiencies associated with opportunistic infections, such as chlamydiosis. Here we characterize a KoRV newly isolated from an animal in a German zoo and show infection of human and rat cell lines in vitro and of rats in vivo, using immunological and PCR methods for virus detection. The KoRV transmembrane envelope protein (p15E) was cloned and expressed, and p15E-specific neutralizing antibodies able to prevent virus infection in vitro were developed. Finally, evidence for immunosuppressive properties of the KoRV was obtained. PMID- 16699048 TI - Rapid upregulation of interferon-regulated and chemokine mRNAs upon injection of 108 international units, but not lower doses, of adenoviral vectors into the brain. AB - The innate immune response, characterized by the rapid induction of proinflammatory genes, plays an important role in immune responses to viral vectors utilized in gene therapy. We demonstrate that several innate proinflammatory mRNAs, i.e., those coding for the interferon (IFN)-regulated proteins interferon regulatory factor 1, 2',5'-oligoadenylate synthetase, and double-stranded-RNA-dependent protein kinase as well as those coding for the chemokines RANTES, IFN-gamma-inducible protein 10, and monocyte chemoattractant protein 1, were all increased in a statistically significant manner in response to 1 x 10(8) IU, but not lower doses, of a first-generation adenovirus injected into the naive brain. This indicates the presence of a threshold dosage of adenovirus needed to elicit an acute innate inflammatory response. PMID- 16699049 TI - Differential drug resistance to anti-tumour necrosis factor agents in rheumatoid arthritis. PMID- 16699050 TI - Lymphoma and other malignancies in primary Sjogren's syndrome. PMID- 16699051 TI - Decreased density of serotonin 5-HT2A receptors in rheumatoid arthritis. AB - BACKGROUND: Animal studies have indicated that 5-HT2A receptors could play a role in arthritic diseases. OBJECTIVE: To analyse the binding properties of 5-HT2A receptors in patients with rheumatoid arthritis. METHODS: Using a radioactive binding assay, 43 patients with rheumatoid arthritis were compared with 49 sex and age matched controls for density and affinity (measured as Bmax and Kd) of 5 HT2A serotonin receptors. Genotyping, using polymerase chain reaction, was undertaken to exclude the possibility that differences in the genetic polymorphism T102C for the 5-HT2A receptor determine differences in receptor density. RESULTS: Mean of Bmax of 5-HT2A receptors in rheumatoid patients was significantly lower than in controls, at 45.3 v 57.4 fmol/mg protein (p = 0.004), but there was no significant difference in Kd. The T102C receptor polymorphism genotypes showed a skewed distribution between the two groups. Even when adjusted for this, there was a significant difference in Bmax between the groups. CONCLUSIONS: The density of 5-HT2A serotonin receptors in patients with rheumatoid arthritis is markedly reduced. This could either reflect a difference involved in the susceptibility to the disease or be a secondary effect of the disease. PMID- 16699052 TI - Facioskeletal changes in children with juvenile idiopathic arthritis. AB - OBJECTIVE: To investigate the facioskeletal morphology in patients with juvenile idiopathic arthritis (JIA) with and without temporomandibular joint (TMJ) involvement. METHODS: Eighty five patients were included. TMJ involvement was defined by orthopantomogram alterations. Lateral cephalograms were used to determine linear and angular measurements and occlusion. RESULTS: Patients regardless of their TMJ status had a 67% chance for retrognathia and a 52% chance for posterior rotation of the mandible and, respectively, 82% and 58% if TMJ involvement were present. Changes were not uniformly distributed among the different subtypes. CONCLUSION: Patients with JIA have an altered facial morphology, especially in the presence of TMJ involvement. PMID- 16699053 TI - Temporal artery biopsy for diagnosing giant cell arteritis: the longer, the better? AB - OBJECTIVE: To investigate the relation between temporal artery biopsy (TAB) length and diagnostic sensitivity for giant cell arteritis. METHODS: Histological TAB reports generated from four hospital pathology departments were reviewed for demographics, histological findings, and formalin fixed TAB lengths. A biopsy was considered positive for giant cell arteritis if there was a mononuclear cell infiltrate predominating at the media-intima junction or in the media. RESULTS: Among 1821 TAB reports reviewed, 287 (15.8%) were excluded because of missing data, sampling errors, or age < 50 years. Mean TAB length of the 1520 datasets finally analysed (67.2% women; mean (SD) age, 73.1 (10.0) years) was 1.33 (0.73) cm. Histological evidence of giant cell arteritis was found in 223 specimens (14.7%), among which 164 (73.5%) contained giant cells. Statistical analyses, including piecewise logistic regression, identified 0.5 cm as the TAB length change point for diagnostic sensitivity. Compared with TAB length of < 0.5 cm, the respective odds ratios for positive TAB without and with multinucleated giant cells in samples > or = 0.5 cm long were 5.7 (95% confidence interval, 1.4 to 23.6) and 4.0 (0.97 to 16.5). CONCLUSIONS: A fixed TAB length of at least 0.5 cm could be sufficient to make a histological diagnosis of giant cell arteritis. PMID- 16699054 TI - Raised serum S100B protein levels in neuropsychiatric lupus. AB - OBJECTIVE: To test serum S100B protein levels in patients with and without neuropsychiatric systemic lupus erythematosus (NPSLE) and controls. METHODS: 87 patients with SLE, 23 with and 64 without neuropsychiatric involvement, and 25 control subjects were prospectively evaluated. NPSLE diagnosis was made according to the American College of Rheumatology nomenclature and case definitions for neuropsychiatric lupus syndromes. Serum S100B protein levels were determined with a luminescence immunoassay. Statistical analysis was performed using Mann-Whitney and Kruskal-Wallis tests. RESULTS: Among the patients with NPSLE, 9 presented psychosis; 4, cranial neuropathy; 3, cerebrovascular disease; 1, seizures; 1, chorea; 1, peripheral polyneuropathy; 1, multiplex mononeuropathy; 3, dementia. Serum concentrations of S100B protein were significantly higher in patients with NPSLE (median 0.164 ng/ml, interquartile range 0.113-0.332) than in non-NPSLE patients (0.062 ng/ml, 0.026-0.109) and controls (0.088 ng/ml, 0.013-0.124) (p<0.001). Patients with anti-dsDNA antibodies had higher S100B protein levels (p = 0.001). No significant associations were found of lupus activity (among non NPSLE cases), antiphospholipid antibodies, and reduced complement levels with S100B concentration. CONCLUSIONS: Serum S100B protein level is raised in NPSLE, reflecting continuing neurological damage. The association of anti-dsDNA antibodies with higher S100B protein concentration deserves further study. PMID- 16699055 TI - Medial tibial stress syndrome due to methotrexate osteopathy. PMID- 16699056 TI - Leflunomide and methotrexate combination therapy in daily clinical practice. PMID- 16699057 TI - Fatal exacerbation of fibrosing alveolitis associated with systemic sclerosis in a patient treated with adalimumab. PMID- 16699058 TI - Effects of acute administration of iloprost on the cardiac autonomic nervous system and ventricular repolarisation in patients with systemic sclerosis. PMID- 16699059 TI - Subchondral insufficiency fracture of the femoral head in a patient with systemic lupus erythematosus. PMID- 16699060 TI - Eosinophilic cellulitis (Wells' syndrome) as a cutaneous reaction to the administration of adalimumab. PMID- 16699061 TI - Genes and schizophrenia: beyond schizophrenia: the role of DISC1 in major mental illness. AB - Schizophrenia and related disorders have a major genetic component, but despite much effort and many claims, few genes have been consistently replicated and fewer have biological support. One recent exception is "Disrupted in Schizophrenia 1" (DISC1), which was identified at the breakpoint on chromosome 1 of the balanced translocation (1;11)(q42.1;q14.3) that co-segregated in a large Scottish family with a wide spectrum of major mental illnesses. Since then, genetic analysis has implicated DISC1 in schizophrenia, schizoaffective disorder, bipolar affective disorder, and major depression. Importantly, evidence is emerging from genetic studies for a causal relationship between DISC1 and directly measurable trait variables such as working memory, cognitive aging, and decreased gray matter volume in the prefrontal cortex, abnormalities in hippocampal structure and function, and reduction in the amplitude of the P300 event-related potential. Further, DISC1 binds a number of proteins known to be involved in essential processes of neuronal function, including neuronal migration, neurite outgrowth, cytoskeletal modulation, and signal transduction. Thus, both genetic and functional data provide evidence for a critical role for DISC1 in schizophrenia and related disorders, supporting the neurodevelopmental hypothesis for the molecular pathogenesis of these devastating illnesses. PMID- 16699063 TI - They only see it when the sun shines in my ears: exploring perceptions of adolescent hearing aid users. AB - Hard-of-hearing (HOH) young people may encounter multiple challenges to their educational, social, and emotional development. The benefits of wearing hearing aids to enhance communication may be countered by negative stigma associated with hearing aids. This study explored the experience of 16 bilaterally, moderately to severely HOH adolescents in mainstream education to understand their perceptions of using hearing aids. A core category of "normality" emerged from the data, which captures the predominant issue for these young people. The sense of being normal may be situationally determined, but those who are able to perceive their use of hearing aids in a given context as normal most frequently use them. Regardless of the age of diagnosis or length of time since fitting, if use of hearing aids is perceived as not normal then their use is disguised or negated. The perceptions of the young people indicate that psychosocial supports focused on promoting their identity as HOH young people and normalizing the use of hearing aids should be a key feature of interventions. PMID- 16699062 TI - Environmental factors in schizophrenia: the role of migrant studies. AB - There is now compelling evidence that migrant groups in several countries have an elevated risk of developing schizophrenia and other psychotic disorders. Though the findings of earlier studies were greeted with skepticism, and ascribed by some to have methodological shortcomings and diagnostic biases, the more rigorous recent studies, from a variety of countries, have still found markedly increased incidence rates. While this phenomenon is an important health issue in its own right, understanding the reasons for the increased rates may provide valuable insights into the causes of schizophrenia and other psychotic disorders in general. The challenge for the next phase of studies is to identify the relevant risk factors and how they might interact to increase the risk of psychosis, both in migrant groups and in the general population. PMID- 16699064 TI - A compelling desire for deafness. AB - A case is described of a patient who has a compelling and persistent desire to become deaf. She often kept cotton wool moistened with oil in her ears and was learning sign language. Living without sound appeared to be a severe form of avoidance behavior from hyperacusis and misophonia. She had a borderline personality disorder that was associated with a poor sense of self. Her desire to be deaf may be one aspect of gaining an identity for herself and to compensate for feeling like an alien and gaining acceptance in the Deaf community. Will a compelling desire for deafness ever become a recognized mental disorder one day for which hearing patients may be offered elective deafness after a period of assessment and living like a deaf person? Those working in the field of deafness should be aware that individuals may occasionally be seeking elective deafness or self-inflicting deafness to obtain a hearing aid. PMID- 16699065 TI - High-glucose-altered endothelial cell function involves both disruption of cell to-cell connection and enhancement of force development. AB - Vascular endothelial cells (ECs), which regulate vascular tonus, serve as a barrier at the interface of vascular tissue. It is generally believed that alteration of this barrier is correlated with diabetic complications; however, a detailed mechanism has not been elucidated. This study examined alteration of bovine arterial EC functions stimulated by a thromboxane A2 analog (9,11-dideoxy 11 alpha,9 alpha-epoxymethano prostaglandin F(2 alpha); U46619) under normal and high-glucose (HG) conditions. U46619 treatment increased EC layer permeability in a time- and dose-dependent fashion. This response initially disrupted calcium dependent EC-to-EC connections, namely, vascular endothelial cadherin (VE-CaD). Thereafter, EC force development in association with morphological changes was detected employing a reconstituted EC fiber technique, resulting in paracellular hole formation in the EC layer. Thus, we confirmed that U46619-induced enhancement of EC layer permeability involves these sequential steps. Similar trials were performed using a concentration twice that of normal glucose (22.2 mM glucose for 48 h). This treatment significantly enhanced U46619-induced EC layer permeability; furthermore, increases in both rate of VE-CaD disruption and EC fiber contraction were evident. Inhibition of calcium-independent protein kinase C and diacylglycerol kinase indicated that the glucose-dependent increase in VE CaD disruption was mediated by a calcium-independent mechanism. Moreover, EC contraction was regulated by a typical calcium-independent pathway associated with rho kinase and actin stress fiber. Contraction was also enhanced under HG conditions. This investigation revealed that glucose-dependent enhancement of EC layer permeability is related to increases in VE-CaD disruption and EC contraction. Increases in both parameters were mediated by alteration of a calcium-independent pathway. PMID- 16699066 TI - Regulation of kindling epileptogenesis by hippocampal galanin type 1 and type 2 receptors: The effects of subtype-selective agonists and the role of G-protein mediated signaling. AB - The search for antiepileptic drugs that are capable of blocking the progression of epilepsy (epileptogenesis) is an important problem of translational epilepsy research. The neuropeptide galanin effectively suppresses acute seizures. We examined the ability of hippocampal galanin receptor type 1 (GalR1) and type 2 (GalR2) to inhibit kindling epileptogenesis and studied signaling cascades that mediate their effects. Wistar rats received 24-h-long intrahippocampal infusion of a GalR1/2 agonist galanin(1-29), GalR1 agonist M617 [galanin(1-13)-Gln14 bradykinin(2-9)-amide], or GalR2 agonist galanin(2-11). The peptides were administered alone or combined with an inhibitor of Gi protein pertussis toxin (PTX), Gi-protein activated K+ channels (GIRK) inhibitor tertiapin Q (TPQ), G(q/11) protein inhibitor [D-Arg1,D-Trp(5,7,9),Leu11]-substance P (dSP), or an inhibitor of intracellular Ca2+ release dantrolene. Sixteen hours into drug delivery, the animals were subjected to rapid kindling-60 electrical trains administered to ventral hippocampus every 5 min. M617 delayed epileptogenesis, whereas galanin(1-29) and galanin(2-11) completely prevented the occurrence of full kindled seizures. TPQ abolished anticonvulsant effect of M617 but not of galanin(2-11). PTX blocked anticonvulsant effects of M617 and inversed the action of galanin(1-29) and galanin(2-11) to proconvulsant. dSP and dantrolene did not modify seizure suppression through GalR1 and GalR2, but eliminated the proconvulsant effect of PTX + galanin(1-29) and PTX + galanin(2-11) combinations. We conclude that hippocampal GalR1 exert their disease-modifying effect through the Gi-GIRK pathway. GalR2 is antiepileptogenic through the Gi mechanism independent of GIRK. A secondary proconvulsant pathway coupled to GalR2 involves G(q/11) and intracellular Ca2+. The data are important for understanding endogenous mechanisms regulating epileptogenesis and for the development of novel antiepileptogenic drugs. PMID- 16699067 TI - Roles of endogenous prostaglandins and cyclooxygenase isozymes in healing of indomethacin-induced small intestinal lesions in rats. AB - The role of prostaglandins (PGs)/cyclooxygenase (COX) in the healing of indomethacin-induced small intestinal ulcers was examined in rats. Animals were given indomethacin (10 mg/kg s.c.) and killed 1, 2, 3, 5, and 7 days later. Indomethacin (2 mg/kg), 5-(4-chlorophenyl)-1-(4-methoxyphenyl)-3 trifluoromethylpyrazole (SC560; COX-1 inhibitor; 3 mg/kg), and rofecoxib (COX-2 inhibitor; 3 mg/kg) were given p.o. once daily for 6 days, during the first 3 days or last 3 days of the experimental period. All COX inhibitors given for 6 days significantly impaired the healing of these ulcers. Healing was also impaired by rofecoxib given for the first 3 days or by SC560 given for the last 3 days. The expression of COX-2 mRNA in the intestine was up-regulated after ulceration, persisting for 3 days and dissipating thereafter. Mucosal PGE2 contents decreased within 3 h after ulceration, recovered 24 h later, and increased above normal 1 approximately 3 days later. The PGE2 content at 4 days after ulceration was decreased by rofecoxib but not SC560, whereas that at 7 days was suppressed by SC560 but not rofecoxib. Vascular content in the ulcerated mucosa decreased when the healing was impaired by COX inhibitors. The deleterious effect of indomethacin on healing was mimicked by a prostacyclin E receptor (EP) 4 antagonist and reversed by coadministration of PGE2 as well as an EP4 agonist. In conclusion, endogenous PGs play a role in the healing of intestinal ulcers through EP4 receptors, yet the COX isozyme involved differs depending on the stage of healing; COX-2 in the early stage and COX-1 in the late stage. PMID- 16699068 TI - An orally active cathepsin K inhibitor, furan-2-carboxylic acid, 1-{1-[4-fluoro-2 (2-oxo-pyrrolidin-1-yl)-phenyl]-3-oxo-piperidin-4-ylcarbamoyl}-cyclohexyl)-amide (OST-4077), inhibits osteoclast activity in vitro and bone loss in ovariectomized rats. AB - Human cathepsin K, a cysteine proteinase of the papain family, has been recognized as a potential drug target for the treatment of osteoporosis. The predominant expression of cathepsin K in osteoclasts has rendered the enzyme into a major target for the development of novel antiresorptive drugs. Now, we report the pharmacological properties of OST-4077 [furan-2-carboxylic acid (1-{1-[4 fluoro-2-(2-oxo-pyrrolidin-1-yl)-phenyl]-3-oxo-piperidin-4-ylcarbamoyl} cyclohexyl)-amide] as a novel selective cathepsin K inhibitor. Human and rat cathepsin K were inhibited in vitro by OST-4077 with the IC50 values of 11 and 427 nM, respectively. OST-4077 suppressed bone resorption induced by rabbit osteoclasts (IC50, 37 nM) but did not affect bone mineralization or cellular alkaline phosphatase activity in MC3T3-E1 cells. Parathyroid hormone-induced bone resorption was inhibited in a dose-dependent manner in thyroparathyroidectomized rats gavaged with a single dose of OST-4077 (ED50, 69 mg/kg). When given orally twice daily for 4 weeks to 3-month-old ovariectomized (OVX) rats, OST-4077 dose dependently prevented bone loss, as monitored by bone densitometry, ash content, and urinary excretion of deoxypyridinoline. No change in serum osteocalcin in the OVX rats by OST-4077 suggested that bone formation might not be affected by the agent. In summary, OST-4077 selectively inhibited bone resorbing activities of osteoclasts and prevented bone loss induced by estrogen deficiency but did not affect bone formation. OST-4077, an orally active selective human cathepsin K inhibitor, may have the therapeutic potential for the treatment of diseases characterized by excessive bone loss including osteoporosis. PMID- 16699069 TI - Cardiac ischemia-reperfusion injury induces matrix metalloproteinase-2 expression through the AP-1 components FosB and JunB. AB - Matrix metalloproteinase-2 (MMP-2) is a central component of the response to injury in the heart. During ischemia, MMP-2 influences ventricular performance and is a determinant of postinfarction remodeling. Elevation of MMP-2 during reperfusion after ischemia suggests that new protein is synthesized, but the molecular regulation of MMP-2 generation during ischemia-reperfusion (I/R) injury has not been studied. Using the MMP-2 promoter linked to a beta-galactosidase reporter in transgenic mice, we investigated the transcriptional regulation and cellular sources of MMP-2 in isolated, perfused mouse hearts subjected to acute global I/R injury. I/R injury induced a rapid activation of MMP-2 promoter activity with the appearance of beta-galactosidase antigen in cardiomyocytes, fibroblasts, and endothelial cells. Activation of intrinsic MMP-2 transcription and translation was confirmed by real-time PCR and quantitative Western blot analyses. MMP-2 transcription and translation were inhibited by perfusion with 1.0 mM hydroxyl radical scavenger N-(-2-mercaptopropionyl)-glycine. Nuclear extracts demonstrated increased abundance of two activator proteins-1 (AP-1) components JunB and FosB following I/R injury. Immunohistochemical staining localized JunB and FosB proteins to the nuclei of all three cardiac cell types following I/R injury, consistent with enhanced nuclear transport of these transcription factors. Chromatin immunoprecipitation (ChIP) of the AP-1 binding site in the intrinsic murine MMP-2 promoter yielded only JunB under control conditions, whereas ChIP following I/R injury recovered both JunB and FosB, consistent with a change in occupancy from JunB homodimers in controls to JunB/FosB heterodimers following I/R injury. We conclude that enhanced MMP-2 transcription and translation following I/R injury are mediated by induction, via oxidant stress, of discrete AP-1 transcription factor components. PMID- 16699070 TI - Increases in diastolic [Ca2+] can contribute to positive inotropy in guinea pig ventricular myocytes in the absence of changes in amplitudes of Ca2+ transients. AB - Increases in contraction amplitude following rest or in elevated extracellular Ca(2+) concentration ([Ca(2+)]) have been attributed to increased sarcoplasmic reticulum (SR) Ca(2+) stores and/or increased trigger Ca(2+). However, either manipulation also may elevate diastolic [Ca(2+)]. The objective of this study was to determine whether elevation of diastolic [Ca(2+)] could contribute to positive inotropy in isolated ventricular myocytes. Voltage-clamp experiments were conducted with high-resistance microelectrodes in isolated myocytes at 37 degrees C. Intracellular free [Ca(2+)] was measured with fura-2, and cell shortening was measured with an edge detector. SR Ca(2+) stores were assessed with 10 mM caffeine (0 mM Na(+), 0 mM Ca(2+)). Following a period of rest, cells were activated with trains of pulses, which generated contractions of increasing amplitude, called positive staircases. Positive staircases were accompanied by increasing diastolic [Ca(2+)] but no change in Ca(2+) transient amplitudes. When extracellular [Ca(2+)] was elevated from 2.0 to 5.0 mM, resting intracellular [Ca(2+)] increased and resting cell length decreased. Amplitudes of contractions and L-type Ca(2+) current increased in elevated extracellular [Ca(2+)], although SR Ca(2+) stores, assessed by rapid application of caffeine, did not increase. Although Ca(2+) transient amplitude did not increase in 5.0 mM extracellular [Ca(2+)], diastolic [Ca(2+)] continued to increase with increasing extracellular [Ca(2+)]. These data suggest that increased diastolic [Ca(2+)] contributes to positive inotropy following rest or with increasing extracellular [Ca(2+)] in guinea pig ventricular myocytes. PMID- 16699071 TI - Competitive displacement of phosphoinositide 3-kinase from beta-adrenergic receptor kinase-1 improves postinfarction adverse myocardial remodeling. AB - Adverse remodeling after myocardial infarction (MI) determines the progression of heart failure. Failing hearts are characterized by downregulation of beta adrenergic receptor (beta-AR) signaling in part because of increased beta-AR kinase 1 activity. Our previous studies have shown that overexpression of the phosphoinositide kinase (PIK) domain of phosphoinositide 3-kinase (PI3K), prevents beta-AR downregulation and enhances adrenergic agonist responsiveness by inhibiting the targeting of PI3K to the beta-AR complex. To investigate whether preventing beta-AR downregulation in the heart ameliorates cardiac function post MI, transgenic mice with cardiac-specific overexpression of the PIK domain peptide (TgPIK) underwent left coronary artery ligation and were subsequently followed by serial echocardiography at 4, 8, 12, 16, and 20 wk. Despite having similar infarction sizes, TgPIK mice showed better systolic function, less cardiac dilatation, and improved hemodynamic response to dobutamine compared with littermate controls after MI. To test that displacement of PI3K from the beta-AR complex, but not the total loss of PI3K-gamma, is critical for amelioration of cardiac function, mice lacking the PI3K-gamma (PI3K-gamma-KO) underwent MI, and their cardiac function was assessed 20 wk post-MI. Serial echocardiographic measurements showed severe reduction in contractile performance in PI3K-gamma-KO compared with TgPIK mice. Furthermore, significant beta-AR downregulation and desensitization were only seen in infarcted wild-type and PI3K-gamma-KO mice and not in TgPIK mice. Together, these results demonstrate that adverse remodeling of the ventricle after MI can be attenuated by a strategy that prevents recruitment of PI3K to the plasma membrane and restores normal beta-AR function. PMID- 16699072 TI - Impact of physiological variables and genetic background on myocardial frequency resistivity relations in the intact beating murine heart. AB - Conductance measurements for generation of an instantaneous left ventricular (LV) volume signal in the mouse are limited, because the volume signal is a combination of blood and LV muscle, and only the blood signal is desired. We have developed a conductance system that operates at two simultaneous frequencies to identify and remove the myocardial contribution to the instantaneous volume signal. This system is based on the observation that myocardial resistivity varies with frequency, whereas blood resistivity does not. For calculation of LV blood volume with the dual-frequency conductance system in mice, in vivo murine myocardial resistivity was measured and combined with an analytic approach. The goals of the present study were to identify and minimize the sources of error in the measurement of myocardial resistivity to enhance the accuracy of the dual frequency conductance system. We extended these findings to a gene-altered mouse model to determine the impact of measured myocardial resistivity on the calculation of LV pressure-volume relations. We examined the impact of temperature, timing of the measurement during the cardiac cycle, breeding strain, anisotropy, and intrameasurement and interanimal variability on the measurement of intact murine myocardial resistivity. Applying this knowledge to diabetic and nondiabetic 11- and 20- to 24-wk-old mice, we demonstrated differences in myocardial resistivity at low frequencies, enhancement of LV systolic function at 11 wk and LV dilation at 20-24 wk, and histological and electron-microscopic studies demonstrating greater glycogen deposition in the diabetic mice. This study demonstrated the accurate technique of measuring myocardial resistivity and its impact on the determination of LV pressure-volume relations in gene-altered mice. PMID- 16699074 TI - Quantification of right ventricular afterload in patients with and without pulmonary hypertension. AB - Right ventricular (RV) afterload is commonly defined as pulmonary vascular resistance, but this does not reflect the afterload to pulsatile flow. The purpose of this study was to quantify RV afterload more completely in patients with and without pulmonary hypertension (PH) using a three-element windkessel model. The model consists of peripheral resistance (R), pulmonary arterial compliance (C), and characteristic impedance (Z). Using pulmonary artery pressure from right-heart catheterization and pulmonary artery flow from MRI velocity quantification, we estimated the windkessel parameters in patients with chronic thromboembolic PH (CTEPH; n = 10) and idiopathic pulmonary arterial hypertension (IPAH; n = 9). Patients suspected of PH but in whom PH was not found served as controls (NONPH; n = 10). R and Z were significantly lower and C significantly higher in the NONPH group than in both the CTEPH and IPAH groups (P < 0.001). R and Z were significantly lower in the CTEPH group than in the IPAH group (P < 0.05). The parameters R and C of all patients obeyed the relationship C = 0.75/R (R(2) = 0.77), equivalent to a similar RC time in all patients. Mean pulmonary artery pressure P and C fitted well to C = 69.7/P (i.e., similar pressure dependence in all patients). Our results show that differences in RV afterload among groups with different forms of PH can be quantified with a windkessel model. Furthermore, the data suggest that the RC time and the elastic properties of the large pulmonary arteries remain unchanged in PH. PMID- 16699073 TI - Vascular endothelial growth factor promotes cardiomyocyte differentiation of embryonic stem cells. AB - Embryonic stem cells (ESCs) overexpressing the vascular endothelial growth factor (VEGF) improve cardiac function in mouse models of myocardial ischemia and infarction by mechanisms that are poorly understood. Here we studied the effects of VEGF on cardiomyocyte differentiation of mouse ESCs in vitro. We used flow cytometry to determine the expression of alpha-myosin heavy chain (alpha-MHC), cardiac troponin I (cTn-I), and Nkx2.5 in differentiated ESCs. VEGF (20 ng/ml) significantly enhanced alpha-MHC, cTn-I, and Nkx2.5 expression in differentiated ESCs. Western blot analysis confirmed these findings. We found that VEGF receptor FMS-like tyrosine kinase-1 (Flt-1) and fetal liver kinase-1 (Flk-1) expression increased during ESC differentiation. Antibodies against Flk-1 totally blocked and against Flt-1 partially blocked VEGF-induced NKx2.5-positive-stained cells. The ERK inhibitor PD-098059 abolished VEGF-induced cardiomyocyte differentiation of ESCs. Our results suggest that VEGF promotes cardiomyocyte differentiation predominantly by ERK-mediated Flk-1 activation and, to a lesser extent, by Flt-1 activation. These findings may be of significance for stem cell and growth factor therapies to regenerate failing cardiomyocytes. PMID- 16699075 TI - Regulation of alpha1-adrenoceptor-mediated contractions of uterine arteries by PKC: effect of pregnancy. AB - Protein kinase C (PKC) plays an important role in the regulation of uterine artery contractility and its adaptation to pregnancy. The present study tested the hypothesis that PKC differentially regulates alpha(1)-adrenoceptor-mediated contractions of uterine arteries isolated from nonpregnant (NPUA) and near-term pregnant (PUA) sheep. Phenylephrine-induced contractions of NPUA and PUA sheep were determined in the absence or presence of the PKC activator phorbol 12,13 dibutyrate (PDBu). In NPUA sheep, PDBu produced a concentration-dependent potentiation of phenylephrine-induced contractions and shifted the dose-response curve to the left. In contrast, in PUA sheep, PDBu significantly inhibited phenylephrine-induced contractions and decreased their maximum response. Simultaneous measurement of contractions and intracellular free Ca(2+) concentrations ([Ca(2+)](i)) in the same tissues revealed that PDBu inhibited phenylephrine-induced [Ca(2+)](i) and contractions in PUA sheep. In NPUA sheep, PDBu increased phenylephrine-induced contractions without changing [Ca(2+)](i). Western blot analysis showed six PKC isozymes, alpha, beta(I), beta(II), delta, epsilon, and zeta, in uterine arteries, among which beta(I), beta(II), and zeta isozymes were significantly increased in PUA sheep. In contrast, PKC-alpha was decreased in PUA sheep. In addition, analysis of subcellular distribution revealed a significant decrease in the particulate-to-cytosolic ratio of PKC epsilon in PUA compared with that in NPUA sheep. The results suggest that pregnancy induces a reversal of PKC regulatory role on alpha(1)-adrenoceptor mediated contractions from a potentiation in NPUA sheep to an inhibition in PUA sheep. The differential expression of PKC isozymes and their subcellular distribution in uterine arteries appears to play an important role in the regulation of Ca(2+) mobilization and Ca(2+) sensitivity in alpha(1)-adrenoceptor mediated contractions and their adaptation to pregnancy. PMID- 16699076 TI - KCa+ channels contribute to exercise-induced coronary vasodilation in swine. AB - Coronary blood flow is controlled via several vasoactive mediators that exert their effect on coronary resistance vessel tone through activation of K(+) channels in vascular smooth muscle. Because Ca(2+)-activated K(+) (K(Ca)(+)) channels are the predominant K(+) channels in the coronary vasculature, we hypothesized that K(Ca)(+) channel activation contributes to exercise-induced coronary vasodilation. In view of previous observations that ATP-sensitive K(+) (K(ATP)(+)) channels contribute, in particular, to resting coronary resistance vessel tone, we additionally investigated the integrated control of coronary tone by K(Ca)(+) and K(ATP)(+) channels. For this purpose, the effect of K(Ca)(+) blockade with tetraethylammonium (TEA, 20 mg/kg iv) on coronary vasomotor tone was assessed in the absence and presence of K(ATP)(+) channel blockade with glibenclamide (3 mg/kg iv) in chronically instrumented swine at rest and during treadmill exercise. During exercise, myocardial O(2) delivery increased commensurately with the increase in myocardial O(2) consumption, so that myocardial O(2) extraction and coronary venous Po(2) (Pcv(O(2))) were maintained constant. TEA (in a dose that had no effect on K(ATP)(+) channels) had a small effect on the myocardial O(2) balance at rest and blunted the exercise-induced increase in myocardial O(2) delivery, resulting in a progressive decrease of Pcv(O(2)) with increasing exercise intensity. Conversely, at rest glibenclamide caused a marked decrease in Pcv(O(2)) that waned at higher exercise levels. Combined K(Ca)(+) and K(ATP)(+) channel blockade resulted in coronary vasoconstriction at rest that was similar to that caused by glibenclamide alone and that was maintained during exercise, suggesting that K(Ca)(+) and K(ATP)(+) channels act in a linear additive fashion. In conclusion, K(Ca)(+) channel activation contributes to the metabolic coronary vasodilation that occurs during exercise. Furthermore, in swine K(Ca)(+) and K(ATP)(+) channels contribute to coronary resistance vessel control in a linear additive fashion. PMID- 16699077 TI - Ventricular-arterial coupling in a rat model of reduced arterial compliance provoked by hypervitaminosis D and nicotine. AB - The vitamin D(3) and nicotine (VDN) model is one of isolated systolic hypertension (ISH) in which arterial calcification raises arterial stiffness and vascular impedance. The effects of VDN treatment on arterial and cardiac hemodynamics have been investigated; however, a complete analysis of ventricular arterial interaction is lacking. Wistar rats were treated with VDN (VDN group, n = 9), and a control group (n = 10) was included without the VDN. At week 8, invasive indexes of cardiac function were obtained using a conductance catheter. Simultaneously, aortic pressure and flow were measured to derive vascular impedance and characterize ventricular-vascular interaction. VDN caused significant increases in systolic (138 +/- 6 vs. 116 +/- 13 mmHg, P < 0.01) and pulse (42 +/- 10 vs. 26 +/- 4 mmHg, P < 0.01) pressures with respect to control. Total arterial compliance decreased (0.12 +/- 0.08 vs. 0.21 +/- 0.04 ml/mmHg in control, P < 0.05), and pulse wave velocity increased significantly (8.8 +/- 2.5 vs. 5.1 +/- 2.0 m/s in control, P < 0.05). The arterial elastance and end systolic elastance rose significantly in the VDN group (P < 0.05). Wave reflection was augmented in the VDN group, as reflected by the increase in the wave reflection coefficient (0.63 +/- 0.06 vs. 0.52 +/- 0.05 in control, P < 0.05) and the amplitude of the reflected pressure wave (13.3 +/- 3.1 vs. 8.4 +/- 1.0 mmHg in control, P < 0.05). We studied ventricular-arterial coupling in a VDN induced rat model of reduced arterial compliance. The VDN treatment led to development of ISH and provoked alterations in cardiac function, arterial impedance, arterial function, and ventricular-arterial interaction, which in many aspects are similar to effects of an aged and stiffened arterial tree. PMID- 16699078 TI - Timing of muscle activation in a hand movement sequence. AB - Recent studies have described muscle synergies as overlapping, multimuscle groups defined by synchronous covariation in activation intensity. A different approach regards a synergy as a fixed temporal sequence of bursts of activity across groups of motoneurons. To pursue this latter definition, the present study used a principal component (PC) analysis tailored to reveal the across-muscle temporal synergies of human hand movement. Electromyographic (EMG) activity was recorded as subjects used a manual alphabet to spell a list of words. The analysis was applied to the EMG waveforms from 27 letter-to-letter transitions of equal duration. The first PC (of 27) represented the main temporal synergy; after practice, it began to account for more of the EMG variance (up to 40%). This main synergy began with a burst in the 4-finger extensor and a silent period in the flexors. There were then progressively later and shorter bursts in the thumb abductor, thumb flexor, little finger abductor, and finally the finger flexors. The results suggest that hand movements may be generated by activity waves unfolding in time. Because finger muscles are under relatively direct cortical control, this suggests a specific form of cortical pattern generation. PMID- 16699079 TI - Effects of focal frontal lesions on response inhibition. AB - This study examined the performance of 38 normal subjects and 43 patients with focal lesions of the frontal lobes on a simple go-nogo task where the probability of the nogo stimulus was either 75% or 25%. Patients with lesions to the superior medial parts of the frontal lobes, in particular to the left superior portion of Brodmann area 6 (which includes the supplementary motor areas and the premotor areas for the right hand) had an increased number of false alarms (incorrect responses to the nogo stimulus). These results indicate that area 6 is specifically involved in the inhibition of response. Patients with lesions to the right anterior cingulate (areas 24 and 32) were slower and more variable in their reaction time. These findings could be explained by an inability to sustain stimulus-response contingencies. Lesions to the right ventrolateral prefrontal cortex (Brodmann areas 44, 45, 47) also increased the variability of response, perhaps by disrupting monitoring performance. PMID- 16699080 TI - Extrinsic and intrinsic systems in the posterior cortex of the human brain revealed during natural sensory stimulation. AB - When exposing subjects to a continuous segment of an audiovisual movie, a large expanse of human cortex, especially in the posterior half of the cerebral cortex, shows stimulus-driven activity. However, embedded within this widespread activity, there are cortical regions whose activity is dissociated from the external stimulation. These regions are intercorrelated among themselves, forming a functional network, which largely overlaps with cortical areas previously shown to be deactivated by task-oriented paradigms. Moreover, the network of areas whose neuronal dynamics are associated with external inputs and the network of areas that appears to be intrinsically driven complement each other, providing coverage of most of the posterior cortex. Thus, we propose that naturalistic stimuli reveal a fundamental neuroanatomical partition of the human posterior cortex into 2 global networks: an "extrinsic" system, comprising areas associated with the processing of external inputs, and an "intrinsic" system, largely overlapping with the task-negative, default-mode network, comprising areas associated with--as yet not fully understood--intrinsically oriented functions. PMID- 16699081 TI - Similar frontal and distinct posterior cortical regions mediate visual and auditory perceptual awareness. AB - Activity in ventral visual cortex is a consistent neural correlate of visual consciousness. However, activity in this area seems insufficient to produce awareness without additional involvement of frontoparietal regions. To test the generality of the frontoparietal response, neural correlates of auditory awareness were investigated in a paradigm that previously has revealed frontoparietal activity during conscious visual perception. A within-experiment comparison showed that frontal regions were related to both visual and auditory awareness, whereas parietal activity was correlated with visual awareness and superior temporal activity with auditory awareness. These results indicate that frontal regions interact with specific posterior regions to produce awareness in different sensory modalities. PMID- 16699082 TI - Neural correlates of processing valence and arousal in affective words. AB - Psychological frameworks conceptualize emotion along 2 dimensions, "valence" and "arousal." Arousal invokes a single axis of intensity increasing from neutral to maximally arousing. Valence can be described variously as a bipolar continuum, as independent positive and negative dimensions, or as hedonic value (distance from neutral). In this study, we used functional magnetic resonance imaging to characterize neural activity correlating with arousal and with distinct models of valence during presentation of affective word stimuli. Our results extend observations in the chemosensory domain suggesting a double dissociation in which subregions of orbitofrontal cortex process valence, whereas amygdala preferentially processes arousal. In addition, our data support the physiological validity of descriptions of valence along independent axes or as absolute distance from neutral but fail to support the validity of descriptions of valence along a bipolar continuum. PMID- 16699083 TI - Associative encoding in anterior piriform cortex versus orbitofrontal cortex during odor discrimination and reversal learning. AB - Recent proposals have conceptualized piriform cortex as an association cortex, capable of integrating incoming olfactory information with descending input from higher order associative regions such as orbitofrontal cortex (OFC). If true, encoding in piriform cortex should reflect associative features prominent in these areas during associative learning involving olfactory cues. To test this hypothesis, we recorded from neurons in OFC and anatomically related parts of the anterior piriform cortex (APC) in rats, learning and reversing novel odor discriminations. Findings in OFC were similar to what we have reported previously, with nearly all the cue-selective neurons exhibiting substantial plasticity during learning and reversal. Also, many of the cue-selective neurons were originally responsive in anticipation of the outcomes early in learning, thereby providing a single-unit representation of the cue-outcome associations. Some of these features were also evident in firing activity in APC, including some plasticity across learning and reversal. However, APC neurons failed to reverse cue selectivity when the associated outcome was changed, and the cue selective population did not include neurons that were active prior to outcome delivery. Thus, although representations in APC are substantially more associative than expected in a purely sensory region, they do appear to be somewhat more constrained by the sensory features of the odor cues than representations in downstream areas of OFC. PMID- 16699084 TI - State-dependent verapamil block of the cloned human Ca(v)3.1 T-type Ca(2+) channel. AB - Verapamil is a potent phenylalkylamine antihypertensive believed to exert its therapeutic effect primarily by blocking high-voltage-activated L-type calcium channels. It was the first clinically used calcium channel blocker and remains in clinical use, although it has been eclipsed by other calcium channel blockers because of its short half-life and interactions with other channels. In addition to blocking L-type channels, it has been reported to block T-type (low-voltage activated) calcium channels. This type of cross-reactivity is likely to be beneficial in the effective control of blood pressure. Although the interactions of T channels with a number of drugs have been described, the mechanisms by which these agents modulate channel activity are largely unknown. Most calcium channel blockers exhibit state-dependence (i.e., preferential binding to certain channel conformations), but little is known about state-dependent verapamil block of T channels. We stably expressed human Ca(v)3.1 T-type channels in human embryonic kidney 293 cells and studied the state-dependence of the drug with macroscopic and gating currents. Verapamil blocked currents at micromolar concentrations at polarized potentials similar to those reported for L-type channels, although unlike for L-type currents, it did not affect current time course. The drug exhibited use-dependence and significantly slowed the apparent recovery from inactivation. Current inhibition was dependent on potential. This dependence was restricted to negative potentials, although all data were consistent with verapamil binding in the pore. Gating currents were unaffected by verapamil. We propose that verapamil achieves its inhibitory effect via occlusion of the channel pore associated with an open/inactivated conformation of the channel. PMID- 16699086 TI - Letter to the editor. PMID- 16699087 TI - Using simulation models to predict feed intake: phenotypic and genetic relationships between observed and predicted values in cattle. AB - The objectives of this study were to evaluate the accuracy of the Decision Evaluator for the Cattle Industry (DECI) and the Cornell Value Discovery System (CVDS) in predicting individual DMI and to assess the feasibility of using predicted DMI data in genetic evaluations of cattle. Observed individual animal data on the average daily DMI (OFI), ADG, and carcass measurements were obtained from postweaning records of 504 steers from 52 sires (502 with complete data). The experimental data and daily temperature and wind speed data were used as inputs to predict average daily feed DMI (kg) required (feed required; FR) for maintenance, cold stress, and ADG; maintenance and cold stress; ADG; maintenance and ADG; and maintenance alone, with CVDS (CFRmcg, CFRmc, CFRg, CFRmg, and CFRm, respectively) and DECI (DFRmcg, DFRmc, DFRg, DFRmg, and DFRm, respectively). Genetic parameters were estimated by REML using an animal model with age on test as a covariate and with genotype, age of dam, and year as fixed effects. Regression equations for observed on predicted DMI were OFI = 1.27 (SE = 0.27) + 0.83 (SE = 0.04) x CFRmcg [R2 = 0.44, residual SD (s(y.x)) = 0.669 kg/d] and OFI = 1.32 (SE = 0.22) + 0.8 (SE = 0.03) x DFRmcg (R2 = 0.53, s(y.x) = 0.612 kg/d). Heritability of OFI was 0.27 +/- 0.12, and heritabilities ranged from 0.33 +/- 0.12 to 0.41 +/- 0.13 for predicted measures of DMI. Phenotypic and genetic correlations between OFI and CFRmcg, CFRmc, CFRg, CFRmg, CFRm, DFRmcg, DFRmc, DFRg, DFRmg, and DFRm were 0.67, 0.73, 0.41, 0.63, 0.78, 0.73, 0.82, 0.45, 0.77, and 0.86 (P < 0.001 for all phenotypic correlations); and 0.95 +/- 0.07, 0.82 +/- 0.13, 0.89 +/- 0.09, 0.95 +/- 0.07, 0.91 +/- 0.09, 0.96 +/- 0.07, 0.89 +/- 0.09, 0.88 +/- 0.09, 0.96 +/- 0.06, and 0.96 +/- 0.07, respectively. Phenotypic and genetic correlations between CFRmcg and DFRmcg, CFRmc and DFRmc, CFRg and DFRg, CFRmg and DFRmg, and CFRm and DFRm were 0.98, 0.94, 0.99, 0.98, and 0.95 (P < 0.001 for all phenotypic correlations), and 0.99 +/- 0.004, 0.98 +/- 0.017, 0.99 +/- 0.004, 0.99 +/- 0.005, and 0.97 +/- 0.021, respectively. The strong genetic relationships between OFI and CFRmcg, CFRmg, DFRmcg, and DFRmg indicate that these predicted measures of DMI may be used in genetic evaluations and that DM requirements for cold stress may not be needed, thus reducing model complexity. However, high genetic correlations for final weight with OFI, CFRmcg, and DFRmcg suggest that the technology needs to be further evaluated in populations with genetic variance in feed efficiency. PMID- 16699088 TI - No associations between PrP genotypes and reproduction traits in INRA 401 sheep. AB - In this study, the potential association of PrP genotypes with ewe reproductive traits and lamb growth traits was investigated. Data were included on ewes and lambs of the INRA 401 breed from the Bourges-La Sapiniere INRA experimental farm. This breed was chosen because of good breeding (prolificacy and maternal abilities) and carcass qualities, and the large number of available animals with performance records and PrP genotypes. Ewes were categorized into 3 PrP genotype classes: ARR homozygous, ARR heterozygous, and animals without any ARR haplotype. Two analyses differing in the traits considered were carried out. Firstly, the potential association of the PrP genotype of ewes with their reproduction traits (fertility, litter size, and ovulation rate) was studied. The data included 801, 779, and 587 genotyped ewes for fertility, litter size, and ovulation rate, respectively. Secondly, the association of the PrP genotype of the ewes with growth traits of their lambs (birth weight, ADG between 10 and 30 d of age) was investigated. The data included 3,749 and 3,512 lambs for birth weight and ADG, respectively. The different traits were analyzed using an animal model, where the PrP genotype effect was included as a fixed effect. The reproductive traits were modeled under a probit or a threshold approach. The results of this study indicate no evidence of an association between PrP genotypes and reproductive and growth traits. It is unlikely that selection for scrapie resistance will have an effect on the reproductive or performance traits studied in the INRA 401 breed. PMID- 16699089 TI - Effects of sire line, sire, and sex on plasma urea nitrogen, body weight, and backfat thickness in offspring of Duroc and Landrace boars. AB - In pork production, the efficiency of dietary protein (AA) use is low, resulting in urinary excretion of large quantities of nitrogen as urea. Use of AA and formation of urea are under enzymatic regulation, suggesting genetic regulation. The current study examined the effects of sire line, sire, and sex on growth characteristics and plasma urea nitrogen (PUN) concentrations in the offspring of 11 Duroc sires and 11 Landrace sires bred to Yorkshire-Landrace dams. Plasma samples were obtained at approximately 107 (age class = 107 d), 128 (age class = 128 d), and 149 (age class = 149 d) d of age from 511 boars, gilts, and barrows group-penned and fed standard finishing diets. Body weight and backfat (BF, mean of 3 measurements) were recorded at the time of blood sample collection. Sex, age class, and their interaction influenced (P < 0.01) BW, BF, and PUN. Predicted traits (i.e., ADG, BW at 21 wk, average daily change in BF, BF at 21 wk, and the mean of 3 PUN measures) were generated. Means (+/-SD) were: ADG, 888 +/- 204 g; BW at 21 wk, 94.2 +/- 12.5 kg; average daily change in BF, 0.083 +/- 0.052 mm; BF at 21 wk, 13.8 +/- 3.0 mm; and the mean of 3 PUN measures, 16.2 +/- 4.4 mg/dL. Predicted weight traits were influenced (P < 0.05) by sire line, and sex influenced (P < 0.01) all predicted traits. Heritability estimates for PUN at 107, 128, and 149 d of age were 0.35 +/- 0.15, 0.21 +/- 0.13, and 0.16 +/- 0.12, respectively. Phenotypic correlations of PUN with growth and fat traits were low. Genetic correlations of PUN measured at 107 d with growth and fat traits were low. However, genetic correlations of PUN measured at 128 or 149 d with growth and fat traits ranged from 0.81 to 0.95. Determination of PUN, as herein, may be of sufficient precision to allow its use in a selection protocol. Selection of pigs with superior growth performance and low PUN may result in a greater efficiency of dietary nitrogen use and a reduced negative environmental impact. PMID- 16699090 TI - Correlated responses in growth, carcass, and meat quality traits to divergent selection for testosterone production in pigs. AB - The objective of this project was to characterize changes in growth, carcass yield, and meat quality traits in castrates and gilts in response to divergent selection for testosterone production. In generation 21, endogenous testosterone concentrations in Duroc boars of the high (HTL) and low (LTL) testosterone lines averaged 49.0 and 27.8 ng/mL (P < 0.01), respectively. Eight LTL and 10 HTL boars were used to sire 29 LTL and 33 HTL litters. To remove the effects of inbreeding, these same boars were mated to females of a Large White x Landrace composite (WC) to generate 11 WC by LTL litters (WLT) and 23 WC by HTL litters (WHT). Castrates and gilts were then allotted to LTL (n = 53), HTL (n = 61), WLT (n = 102), and WHT (n = 101) for testing. Growth and carcass traits analyzed included days to 114 kg (D114), ADG, backfat adjusted to 114 kg (ABF), LM area adjusted to 114 kg and predicted percent lean (PPL). Fat-O-Meater data collected were adjusted fat depth (AFD), adjusted loin depth, and percent lean. Meat quality traits characterized at 24 h postmortem included marbling score, percent lipid, pH, drip loss, color score, and Minolta L*, a*, and b*. Data were analyzed with a mixed model including fixed effects of line, mating type (purebred or crossbred), sex, and the random effect of sire nested within line. All possible interactions among fixed effects were tested. The HTL had fewer D114 (P < 0.05), greater ADG (P < 0.01), greater ABF (P < 0.01), and lower PPL (P < 0.01) than LTL. The WHT and WLT did not differ for D114, ADG, or ABF. The WHT had smaller LM area adjusted to 114 kg (P < 0.05) and greater drip loss (P < 0.05) than WLT. The WLT had lower adjusted loin depth (P < 0.05) than LTL and HTL. The LTL and HTL had greater subjective scores for marbling (P < 0.05) compared with WLT and WHT. The least squares mean for percent lipid for HTL and LTL was 4.00. The WHT had greater means for L*, a*, and b* (P < 0.05) than WLT. Pigs selected for increased testosterone production grew faster and produced fatter carcasses than pigs selected for decreased testosterone. Changes in growth, carcass yield, and meat quality traits were detected in castrates and gilts in response to divergent selection for testosterone production. PMID- 16699091 TI - The effect of ignoring individual heterogeneity in Weibull log-normal sire frailty models. AB - The objective of this study was, by means of simulation, to quantify the effect of ignoring individual heterogeneity in Weibull sire frailty models on parameter estimates and to address the consequences for genetic inferences. Three simulation studies were evaluated, which included 3 levels of individual heterogeneity combined with 4 levels of censoring (0, 25, 50, or 75%). Data were simulated according to balanced half-sib designs using Weibull log-normal animal frailty models with a normally distributed residual effect on the log-frailty scale. The 12 data sets were analyzed with 2 models: the sire model, equivalent to the animal model used to generate the data (complete sire model), and a corresponding model in which individual heterogeneity in log-frailty was neglected (incomplete sire model). Parameter estimates were obtained from a Bayesian analysis using Gibbs sampling, and also from the software Survival Kit for the incomplete sire model. For the incomplete sire model, the Monte Carlo and Survival Kit parameter estimates were similar. This study established that when unobserved individual heterogeneity was ignored, the parameter estimates that included sire effects were biased toward zero by an amount that depended in magnitude on the level of censoring and the size of the ignored individual heterogeneity. Despite the biased parameter estimates, the ranking of sires, measured by the rank correlations between true and estimated sire effects, was unaffected. In comparison, parameter estimates obtained using complete sire models were consistent with the true values used to simulate the data. Thus, in this study, several issues of concern were demonstrated for the incomplete sire model. PMID- 16699092 TI - True ileal amino acid digestibility and endogenous ileal amino acid losses in growing pigs fed wheat shorts- or casein-based diets. AB - Use of dietary AA in growing pigs reflects digestion and use of digested AA for various body functions. Before evaluating dietary effects on use of digestible AA intake for body protein deposition, a digestibility study was conducted to investigate true ileal AA digestibility and endogenous ileal AA losses in growing pigs fed graded levels of wheat shorts (WS) or casein (CS; control). A casein based basal diet (basal) was formulated to contain 0.27 g of standardized ileal digestible (SID) Lys per MJ of DE, to which extra Lys was added from WS (WS2, +0.10 g of SID Lys per MJ of DE; WS3, +0.20 g of SID Lys per MJ of DE) or casein (CS3, +0.20 g of SID Lys per MJ of DE). A fifth diet was formulated to be similar in CP level and source as CS3 but in which 6% pectin, a source of soluble non starch polysaccharides (NSP), was included at the expense of cornstarch (CS3 + pectin). Five Yorkshire barrows (17.5 +/- 1.5 kg of BW) were fitted with a T cannula at the distal ileum and randomly assigned to 1 of the 5 experimental diets in a 5 x 5 Latin Square design. Apparent ileal digestibility (AID), true ileal digestibility (TID), and endogenous ileal protein losses (EPL) were determined using the homoarginine method. Diet CS level did not influence (P > or = 0.10) TID of most essential AA or EPL (10.4 g/kg of DM intake). Including pectin in the diet did not influence TID of AA (P > or = 0.10) but increased EPL (15.6 g/kg of DM intake; P > or = 0.01). Inclusion of WS in the diet reduced TID of most essential AA (P < 0.01). The TID values for most essential AA, however, were the same (P > or = 0.10) for both dietary WS levels, except for Lys and Met, which were further reduced at the greatest dietary WS level. Increased EPL (P < 0.01) was only observed for WS3 (16 g/kg of DMI). We concluded that (1) the effects of dietary protein source on AID of AA can be attributed both to reduced TID of AA and increased EPL, (2) the impact of dietary WS level on TID of AA and EPL does not seem to be linear, (3) soluble NSP from pectin or WS exerts a greater effect on EPL than insoluble NSP, and (4) because of the metabolic cost associated with EPL and the impacts of feed composition on microbial fermentation in the gut lumen, the effects of feed ingredients on the use of ileal digestible AA for protein deposition should be investigated further. PMID- 16699093 TI - Efficiency of utilizing ileal digestible lysine and threonine for whole body protein deposition in growing pigs is reduced when dietary casein is replaced by wheat shorts. AB - To determine the effect of dietary inclusion level of wheat shorts (WS; a high nonstarch poly-saccharide-containing feed ingredient) and casein (CS; a control) on the efficiency of utilizing ileal digestible Lys (kLys) and Thr (kThr) for whole body protein deposition (PD) in the growing pig, 2 separate N-balance studies were conducted with either Lys or Thr as first-limiting AA in cornstarch based diets. For the Lys study, a basal diet (L-basal) was formulated to contain 0.24 g of standardized ileal digestible (SID) Lys per MJ of DE, to which 0.095 or 0.19 g of SID Lys per MJ of DE were added using either CS (L-CS2 or L-CS3, respectively) or WS (L-WS2 or L-WS3, respectively). A sixth diet was evaluated that was similar to L-CS3 but to which 6% pectin (L-pectin) was added as a source of soluble nonstarch polysaccharides. For the Thr study, the basal diet (T-basal) was formulated to contain 0.14 g of SID Thr per MJ of DE, to which 0.055 or 0.11 g of SID Thr per MJ of DE were added from CS (T-CS2 or T-CS3, respectively) or from WS (T-WS2 and T-WS3, respectively). A sixth diet was evaluated that was similar to T-CS3 but to which 6% pectin was added (T-pectin). Increasing SID Lys intake from CS did not influence kLys for PD (P > 0.10), whereas increasing SID Lys intake from WS reduced kLys for PD (P = 0.001; 89 vs. 79%). Inclusion of 6% pectin had no effect on kLys for PD (P > 0.10). Increasing SID Thr intake from CS also did not influence kThr for PD (P > 0.10), whereas kThr for PD was reduced at the greatest dietary inclusion level of WS (P < 0.001; 90 vs. 77%). Pectin inclusion had no effect on kThr for PD (P > 0.10). The inefficiency of utilizing ileal digestible Lys intake for PD may be attributed to nonreactive Lys in WS. The negative impact of including high levels of WS in the diet of pigs on kThr seems to be associated with fiber content of WS; it was not related to increased endogenous ileal AA losses at the distal ileum. The impact of dietary AA source on the use of ileal digestible Lys and Thr for PD, or other body functions, is substantial and should be considered in the formulation of pig diets. Further research is warranted to elucidate the mechanisms contributing to substantial dietary effects on Thr use for PD. PMID- 16699094 TI - Intestinal morphology and enzymatic activity in newly weaned pigs fed contrasting fiber concentrations and fiber properties. AB - The main objective of this study was to determine the effect of fiber source and concentration on morphological characteristics, mucin staining pattern, and mucosal enzyme activities in the gastrointestinal tract of pigs. The experiment included 50 pigs from 10 litters weaned at 4 wk of age (BW 8.6 +/- 1.4 kg) and divided into 5 treatment groups. Diets containing fiber of various physico chemical properties and concentrations were formulated to contain 73, 104, or 145 g of dietary fiber/kg of DM. The diets were based on raw wheat and barley flours. Pectin and barley hulls, representing soluble and insoluble fiber sources, respectively, were used to increase the fiber concentration. The pigs were fed the experimental diets for 9 d, and then the pigs were euthanized and the entire gastrointestinal tract was removed. Tissue samples were taken from the mid and distal small intestine and from the mid colon. Inclusion of pectin in the diets significantly decreased (P < 0.001) ADFI and ADG compared with pigs fed no pectin. The villi and the crypts were shorter in pigs fed pectin-containing diets, but the villous height/crypt depth ratio was unaltered. Pectin significantly decreased the area of mucins in the crypts of the small intestine, indicating that the pigs fed the pectin-containing diet would probably be more susceptible to pathogenic bacteria, although this cannot be separated from the impact on ADFI. The lectin-binding pattern of the intestinal mucosa was unaffected by diet. The activity of lactase and maltase was increased in pigs fed diets with high fiber content, whereas sucrase activity was increased in pigs fed the pectin-containing diets. The activity of the peptidases, aminopeptidase N and dipeptidylpeptidase IV, was increased when feeding high fiber diets, whereas the activity of gamma-glutamyl transpeptidase remained unaffected by the experimental diets. In conclusion, the reduced feed intake observed with the pectin-containing diets could explain the lower villous height and crypt depth observed in this study. However, direct effects of pectin also are possible, and thus further study is warranted. Feeding pigs high insoluble fiber diets improved gut morphology by increasing villi length and increased mucosal enzyme activity when compared with pigs fed pectin-containing diets. The mucin content as determined by staining characteristics suggests that pigs fed high insoluble fiber diets might be better protected against pathogenic bacteria than pigs fed diets high in soluble fiber. PMID- 16699095 TI - Selecting soybean meal characteristics preferred for swine nutrition. AB - As environmental constraints become more important issues for the animal industry, selecting feed ingredients that yield good animal performance but also minimize environmental impact of animal production becomes critical. The objective of this research was to identify which compositional features would be desirable for soybean meal to maximize nutritional value and minimize animal waste. Eight soybean samples were selected from a database of 72, such that maximal variability for CP, NDF, and ADF content was obtained. Samples were subsequently processed into meal using standardized procedures. In Experiment 1, 8 cannulated pigs were used to determine ileal digestibility following a Latin square design. In Experiment 2, 5 of the samples were used in complete feeds and 10 pigs were used in a crossover Latin square design to determine the total tract digestibility, odorants in fresh and 5-d-old manure, and ammonia emission from manure. Differences up to 6% in ileal DM digestibility and 8% in ileal CP digestibility were observed. This difference was reduced to 1.1% for total tract DM digestibility and 4% for total tract CP digestibility. Differences in odorant concentration were 3-fold and for in vitro ammonia emission were 42%. The only compositional variable with a significant effect on digestibility was stachyose, which negatively affected ileal digestibility of DM (r = -0.80, P = 0.02) and energy (r = -0.73, P = 0.04). None of the compositional variables measured affected ileal CP digestibility. Ileal CP digestibility, however, was correlated with estimated CP fermentation in the large intestine (r = -0.86, P = 0.06) and with in vitro ammonia emission after 48 h (r = -0.81, P = 0.09). In conclusion, nutritionally relevant variability exists in soy varieties. Low stachyose content is important for maximizing ileal energy digestibility of soybean meal. Although no compositional variable was identified that explained differences in ileal CP digestibility, maximizing ileal CP digestibility is of interest for maximizing the nutritional value of soybean meal and possibly for reducing ammonia and odor emissions. PMID- 16699096 TI - Amino acid and energy digestibility of protein sources for growing pigs. AB - Two experiments were conducted to determine the apparent ileal digestibility (AID) and standardized ileal digestibility (SID) of AA and DE, and to estimate ME and NE of rice protein concentrate, salmon protein hydrolysate, whey protein concentrate, and spray-dried plasma protein. In Exp. 1, 6 barrows (initially 29.5 +/- 2.5 kg of BW) were fitted with ileal T-cannulas and fed each of 5 cornstarch based diets in a balanced crossover design over 35 d. During a given week, there were either 1 or 2 replications of each treatment, resulting in 6 total replications over 5 wk. The 4 test diets (fed from d 0 to 28) were formulated to contain 12.5% CP by using analyzed nutrient compositions of rice protein concentrate, salmon protein hydrolysate, whey protein concentrate, or spray-dried plasma protein. The fifth (N-free) diet was fed from d 28 to 35 to estimate basal endogenous losses of CP and AA, which were used to calculate SID. Ileal digesta were collected and analyzed, and AID and SID values were calculated. Apparent ileal digestible Lys, Met, and Thr values were 80.0 +/- 3.3, 65.6 +/- 3.1, and 68.4 +/- 4.5% for rice protein concentrate; 85.6 +/- 4.8, 85.5 +/- 4.3, and 69.8 +/- 8.5% for salmon protein hydrolysate; 93.3 +/- 1.4, 89.9 +/- 5.8, and 83.6 +/- 5.3% for whey protein concentrate; and 92.8 +/- 0.9, 85.7 +/- 2.1, 86.5 +/- 2.3% for spray-dried plasma protein, respectively. In Exp. 2, 6 barrows (initially 37.6 +/- 1.7 kg of BW) were fed each of 5 corn-based diets in a balanced crossover design over 35 d. During a given week, there were either 1 or 2 replications of each treatment, resulting in 6 total replications over 5 wk. The 4 diets containing the test ingredients were formulated to contain approximately 20% CP by using their analyzed nutrient compositions. The fifth (corn control) diet containing 8.2% CP was also used to calculate energy values by difference. Feces were collected to determine DE. The ME and NE contents were estimated using published regression equations. The DE, ME, and NE (as-fed) values were 4,724 +/- 461, 4,226 +/- 437, and 3,235 +/- 380 kcal/kg for rice protein concentrate; 4,173 +/- 1,052, 3,523 +/- 1,002, and 2,623 +/- 872 kcal/kg for salmon protein hydrolysate; 4,949 +/- 1,002, 4,352 +/- 955, and 3,344 +/- 831 kcal/kg for whey protein concentrate; and 4,546 +/- 673, 3,979 +/- 652, and 3,020 +/- 567 kcal/kg for spray-dried plasma protein, respectively. The excellent AA digestibility and relatively high DE, ME, and NE values indicate that these protein sources warrant further investigation as ingredients for growing pig diets. PMID- 16699097 TI - Evaluation of the inclusion of soybean oil and soybean processing by-products to soybean meal on nutrient composition and digestibility in swine and poultry. AB - This experiment was designed to evaluate the effects of selected soybean (SB) processing byproducts (gums, oil, soapstock, weeds/trash) when added back to soybean meal (SBM) during processing on the resulting nutrient composition, protein quality, nutrient digestibility by swine, and true metabolizable energy (TMEn) content and standardized AA digestibility by poultry. To measure ileal DM and nutrient digestibility, pigs were surgically fitted with a T-cannula in the distal ileum. The concentration of TMEn and the standardized AA digestibility by poultry were determined using the precision fed cecectomized rooster assay. Treatments in the swine experiment included SBM with no by-products; SBM with 1% gum; SBM with 3% gum; SBM with 0.5% soapstock; SBM with 1.5% soapstock; SBM with 2% weeds/trash; SBM with a combination of 3% gum, 1.5% soapstock, and 2% weeds/trash; SBM with 5.4% soybean oil; and roasted SB. A 10 x 10 Latin square design was utilized. The experiment was conducted at the University of Illinois, Urbana-Champaign, and at The Ohio State University, Columbus. In the swine experiment, apparent ileal DM, OM, CP, and AA digestibilities were reduced (P < 0.05) when pigs consumed the combination by-product diet compared with the diet containing no by-products. Apparent ileal digestibilities of DM, CP, and total essential, total nonessential, and total AA were lower (P < 0.05) for any diet containing by-products compared with the diet with no by-products. Apparent ileal digestibilities of DM, OM, CP, and AA were lower (P < 0.05) for the roasted SB compared with the SB oil-containing diet. In the rooster experiment, TMEn values were greater (P < 0.05) for roasted SB compared with SBM with no by-products and increased linearly as the addition of soapstock increased. Individual, total essential, total nonessential, and total AA digestibilities were lower (P < 0.05) for roosters fed roasted SB versus SBM devoid of by-products. Gums, soapstock, and weeds/trash reduce the nutritive value of the resultant meal when they are added back during processing. PMID- 16699098 TI - The dietary valine requirement for prolific lactating sows does not exceed the National Research Council estimate. AB - Two studies were conducted to determine the effect of increasing the valine:lysine (V:L) ratio in diets of lactating sows above the minimum proposed by the NRC (1998). The first experiment involved 189 PIC, Camborough product sows (parity 1 to 4) that were allotted to 1 of 3 dietary treatments. Diets were formulated to achieve total dietary V:L ratios of 0.90, 1.05, or 1.25:1, respectively, and were corn and soybean meal-based. The second experiment involved 279 PIC, Camborough sows (parity 1 to 5) that were allotted to 1 of 4 treatments. Diets 1 and 3 were formulated using corn and a fixed inclusion of soybean meal (16.7%), with 0.27% L-lysine HCl. The V:L ratios in diets 1 and 3 were 0.73 and 1.25:1, respectively. Diets 2 and 4 were typical corn-soybean meal diets containing 0.05% L-lysine HCl, with a fixed inclusion of soybean meal (22.7%). The V:L ratios in diets 2 and 4 were 0.86 and 1.25:1, respectively. In both experiments, each litter was standardized to a minimum of 10 pigs, which achieved litter growth rates of 2.22 and 2.56 kg/d in Exp. 1 and 2, respectively. In Exp. 1, increasing the dietary V:L ratio beyond 0.90:1 did not improve (P > 0.10) the number of pigs weaned, survival rate, or piglet growth rate, even though sows were nursing more than 10 pigs per litter for 19 d. In Exp. 2, total lysine intake was similar among treatments and ranged from 52.1 to 55.3 g/d. Valine intake increased as the diet valine concentration increased (diet 1 vs. 3 and diet 2 vs. 4, P < 0.001), ranging from 40.0 to 66.1 g/d. Litter gain tended to improve (P = 0.14) when the 0.27% L-lysine HCl control (0.73 V:L) was supplemented with valine to achieve a 1.25:1 V:L ratio. In contrast, no aspect of sow or litter response was improved when the practical control diet containing 0.05% L-lysine HCl (0.86 V:L) was supplemented with valine to achieve a 1.25:1 V:L ratio. Collectively, this research shows that a V:L ratio in excess of 0.86 does not conserve maternal tissue loss or improve piglet growth rate, but a V:L ratio of 0.73 may compromise litter growth rate. PMID- 16699099 TI - Effects of supplementation of beta-glucans on growth performance, nutrient digestibility, and immunity in weanling pigs. AB - Two experiments were conducted to evaluate the efficacy of beta-glucan on growth performance, nutrient digestibility, and immunity in weanling pigs. In Exp. 1, 210 weanling pigs (6.38 +/- 0.92 kg of BW) were fed dietary beta-glucan (0, 0.01, 0.02, 0.03, or 0.04%) for 5 wk. In Exp. 2, 168 pigs (6.18 +/- 1.31 kg of BW) were fed no beta-glucan or antibiotics (T1), 0.02% beta-glucan (T2), only antibiotics (T3), or 0.02% beta-glucan with antibiotics (T4) for 8 wk. In Exp. 2, the antibiotics fed were apramycin and carbadox in phase I (0 to 2 wk) and carbadox and chlortetracycline in phase II (3 to 8 wk). During Exp. 2, the performance study was conducted for 5 wk, and the immune response was tested until 8 wk. In Exp. 1, there was a trend for a linear increase (P = 0.068) in ADG as the dietary beta-glucan concentration increased in the diet. The digestibilities of DM, GE, CP, ether extract, Ca, and P increased linearly (P < 0.05) in the beta-glucan supplemented pigs. In Exp. 2, the overall ADG was greater (P < 0.05) in treatment T4 compared with the control group (T1). Also, except for P, this group showed greater (P < 0.05) nutrient digestibilities than the control group. In Exp. 2, at d 15, 24, and 46 antibody titers were measured by ELISA against Pasteurella multocida type A and D after vaccination with atrophic rhinitis, and they differed significantly (P < 0.05) with no particular trend. Flow cytometry was used to determine porcine lymphocyte subpopulations at 4 and 8 wk of Exp. 2. There was an increase in CD4 cells (P < 0.05) and a trend for an increase in CD8 cells (P < 0.10) at 8 wk in pigs fed the T2 diet compared with the other groups. Overall, increasing the dietary concentrations of beta-glucan did not improve ADG without antibiotic, and in weanling pigs antibiotics seem to be more effective in improving nutrient digestibilities and growth performance than beta-glucan. PMID- 16699100 TI - Changes in heat production by mature cows after changes in feeding level. AB - We hypothesized that adaptation of heat production in the realimented cow would occur over an extended period, and the length of time would be influenced by the level of feed. Our objectives were to quantify the changes in heat production of cows after feed restriction and to quantify the effect of level of realimentation on the dynamics of heat production in lightweight cows. Forty 4-yr-old nonpregnant, nonlacting cows (4-breed composite: 1/4 Hereford, 1/4 Angus, 1/4 Red Poll, and 1/4 Pinzgauer) were randomly assigned to receive 1 of 4 levels of a common alfalfa hay source. All cows were feed-restricted [50.0 g of DM/metabolic body size (MBS, kg of BW(0.75)); period 1], and individual fed heat production measurements were taken 0, 7, 13, 28, 56, and 91 d after feed restriction (period 1). In period 2, cows were fed their assigned feed level for their treatment after d 91 of restriction: 50.0 (T50.0), 58.5 (T58.5), 67.0 (T67.0), and 75.5 (T75.5) g of DM/MBS. Measures were taken at 7, 13, 28, 42, 56, 91, 119, and 175 d. In period 3, all cows were fed 75.5 g of DM/MBS after their 175-d measurement, and measures were taken at 7, 14, 28, 56, and 112 d later. In period 1, heat production decreased rapidly during the first 7 d of feed restriction, and heat production continued to decrease during the 91-d restriction. Heat production increased rapidly within the first 7 d, but chronic adaptation continued for T75.5 and T67.0 cows. In period 3, heat production increased rapidly during the first 7 d. Heat production scaled for metabolic body size tended to differ among treatments (P = 0.11). Daily heat production increased by 2.5 kcal/d. These data suggest that there is not a lag in heat production during realimentation and that increased recovered energy is associated with a rapid increase in heat production. PMID- 16699101 TI - Effects of a long-acting, trace mineral, reticulorumen bolus on range cow productivity and trace mineral profiles. AB - The objectives were to determine if strategic supplementation of range cows with a long-acting (6 mo), trace mineral, reticulorumen bolus containing Cu, Se, and Co would: (1) increase cow BCS and BW, and calf birth, weaning, and postweaning weights, or weight per day of age (WDA); (2) increase liver concentrations of Cu or Zn in cows, or blood Se, Cu, or Zn concentrations in cows and calves; and (3) vary by cow breed for any of these response variables. There were 192 control and 144 bolused Composite cows (C; 25% Hereford, Angus, Gelbevieh, and Senepol or Barzona); 236 control and 158 bolused Hereford (H) cows; and 208 control and 149 bolused Brahman cross (B) cows used in a 3-yr experiment. Cows were weighed and scored for body condition in January, May, and September, and all bolused cows received boluses in January. Each year, from among the 3 breed groups a subset of 15 control and 15 bolused cows (n = 90) had samples obtained in January and May for liver Cu and Zn, blood Se, and serum Cu and Zn. As for cows, blood and serum from the calves of these cows were sampled each year in May and September for Cu, Se, and Zn. There was a significant breed x year x treatment interaction (P = 0.001) for cow weight loss from January to May. Calf WDA, weaning, and postweaning weights did not differ (P > 0.40) between bolused and control cows, but there was a significant (P = 0.022) breed x year x treatment interaction for birth weight. Liver Cu was deficient (< 75 ppm; P < 0.001) in control cows and adequate (< 75 to 90 ppm) for bolused cows. Liver Cu differed by year (P < 0.001). Blood Se was adequate (< 0.1 ppm) for all cows except in January 2001 and 2002. There was no difference (P > 0.50) in blood Se between treatment groups in January, but bolused cows had greater (P < 0.01) blood Se in May. Breed differences for blood Se concentrations existed for bolused cows, with B having greater (P < 0.05) blood Se than either C or H cows. Breed differences also existed for control cows, with H having less blood Se (P < 0.04) than B or C cows. Calves from bolused cows had greater blood Se than calves from control cows (P = 0.01). Supplementation via a long-acting trace mineral bolus was successful in increasing liver Cu in cows and blood Se in cows and calves, but the responses varied by year. Bolus administration had variable effects on BW change in early lactation, depending on breed and year, which may indicate the need for breed- and year-specific supplementation programs. PMID- 16699102 TI - Growth and body composition of dairy calves fed milk replacers containing different amounts of protein at two feeding rates. AB - Previous research has demonstrated that increasing the CP concentration from 16 to 26% in milk replacers fed to male preruminant dairy calves at 1.5% of BW (DM basis) daily resulted in increased ADG, G:F, and deposition of lean tissue. However, the effects of dietary CP would be expected to vary depending on ME intake. Here, male Holstein calves < 1 wk old were used to determine the effects of feeding rate and CP concentration of isocaloric, whey protein-based milk replacers on growth and body composition. After a 2-wk standardization period, calves were assigned randomly to an initial baseline group or to treatments in a 2 x 4 factorial arrangement of feeding rate (1.25 or 1.75% of BW daily, DM basis) and milk replacer CP concentration (14, 18, 22, or 26% of DM). No starter was offered, but calves had free access to water. After a 5-wk feeding period, calves were slaughtered and body composition was determined. Increasing the feeding rate increased (P < 0.05) ADG, G:F, empty-body gains of chemical components and energy, the percentage of fat in empty BW gain and in the final empty body, and concentrations of IGF-I and insulin in plasma. Increasing the feeding rate decreased (P < 0.01) percentages of water and protein in the empty body and decreased urea N in plasma. Increasing dietary CP concentration linearly increased (P < 0.05) ADG, body length, heart girth, and gains of water and protein but linearly decreased (P < 0.05) fat gain. As dietary CP increased, fat content in empty body gain decreased, and water and protein increased. Increasing CP concentration increased (quadratic, P < 0.02) G:F, with greatest efficiencies for calves fed 22% CP. Gross energetic efficiency (retained energy:intake energy) was greater (P < 0.05) for calves fed at 1.75% of BW daily. Efficiency of dietary protein use for protein gain was greater for calves fed at 1.75% of BW daily but was not affected by dietary CP. The ratio of protein gain to apparently digestible protein intake above maintenance decreased as dietary CP increased. Interactions (P < 0.05) of feeding rate and CP concentration for gains of water and protein indicated that when dietary CP was 26% the ME supply limited protein use by calves fed at 1.25% of BW daily. Body composition of preruminant calves can be markedly altered by manipulating the protein to energy ratio in milk replacers. These dietary effects on body composition and growth are not predicted by current NRC standards. PMID- 16699103 TI - Effects of corn condensed distillers solubles supplementation on ruminal fermentation, digestion, and in situ disappearance in steers consuming low quality hay. AB - Two metabolism (4 x 4 Latin square design) experiments were conducted to evaluate the effects of corn condensed distillers solubles (CCDS) supplementation on intake, ruminal fermentation, site of digestion, and the in situ disappearance rate of forage in beef steers fed low-quality switchgrass hay (Panicum virgatum L.). Experimental periods for both trials consisted of a 9-d diet adaptation and 5 d of collection. In Exp. 1, 4 ruminally and duodenally cannulated steers (561 +/- 53 kg of initial BW) were fed low-quality switchgrass hay (5.1% CP, 40.3% ADF, 7.5% ash; DM basis) and supplemented with CCDS (15.4% CP, 4.2% fat; DM basis). Treatments included 1) no CCDS; 2) 5% CCDS; 3) 10% CCDS; and 4) 15% CCDS (DM basis), which was offered separately from the hay. In Exp. 2, 4 ruminally and duodenally cannulated steers (266.7 +/- 9.5 kg of initial BW) were assigned to treatments similar to Exp. 1, except forage (Panicum virgatum L.; 3.3% CP, 42.5% ADF, 5.9% ash; DM basis) and CCDS (21.6% CP, 17.4% fat; DM basis) were fed as a mixed ration, using a forage mixer to blend the CCDS with the hay. In Exp. 1, ruminal, postruminal, and total tract OM digestibilities were not affected (P = 0.21 to 0.59) by treatment. Crude protein intake and total tract CP digestibility increased linearly with increasing CCDS (P = 0.001 and 0.009, respectively). Microbial CP synthesis tended (P = 0.11) to increase linearly with increasing CCDS, whereas microbial efficiency was not different (P = 0.38). Supplementation of CCDS to low-quality hay-based diets tended to increase total DM and OM intakes (P = 0.11 and 0.13, respectively) without affecting hay DMI (P = 0.70). In Exp. 2, ruminal OM digestion increased linearly (P = 0.003) with increasing CCDS, whereas postruminal and total tract OM digestibilities were not affected (P > or = 0.37) by treatment. Crude protein intake, total tract CP digestibility, and microbial CP synthesis increased (P < or = 0.06) with increasing level of CCDS supplementation, whereas microbial efficiency did not change (P = 0.43). Ruminal digestion of ADF and NDF increased (P = 0.02 and 0.008, respectively) with CCDS supplementation. Based on this data, CCDS used in Exp. 2 was 86.7% rumen degradable protein. The results indicate that CCDS supplementation improves nutrient availability and use of low-quality forages. PMID- 16699104 TI - Effects of 25-hydroxyvitamin D3 and manipulated dietary cation-anion difference on the tenderness of beef from cull native Korean cows. AB - In this study, we characterized the effects of 25-hydroxyvitamin D3 (25-OH D3) and manipulated dietary cation-anion difference (DCAD) on the performance, urine pH, serum constituents, carcass traits, tissue residual vitamin D and its metabolites, beef tenderness, and mRNA and protein concentrations of Ca-dependent proteinases in LM using 24 cull native Korean cows. The cows were divided into 3 groups of 8: control, 25-OH D3 supplemented (25-OH D3), and manipulated DCAD plus 25-OH D3 supplemented (DCAD+25-OH D3). Cows receiving 25-OH D3 or DCAD+25-OH D3 were dosed with 125 mg of 25-OH D3 6 d before slaughter. The manipulated DCAD ( 10 mEq/100 g of DM) diet was fed from 20 to 6 d (14 d) before slaughter. The DCAD+25-OH D3 treatment decreased urine pH and increased serum Ca concentrations. Although the vitamin D concentrations in LM, liver, and kidney were not affected by 25-OH D3 or DCAD+25-OH D3, muscle tissue 25-OH D3 concentrations were increased by both regimens. Serum 25-OH D3 concentrations were increased by 25-OH D3 supplementation, and the increase was even greater for DCAD+25-OH D3. The same pattern was observed for serum 1,25- (OH)2 D3. However, the LM concentration of 1,25-(OH)2 D3 was less for DCAD+25-OH D3 than for control. Although Ca concentrations of LM increased numerically in response to 25-OH D3 supplementation, no statistical differences in Warner-Bratzler shear force or sensory traits of LM were detected. The LM of cows receiving 25-OH D3 with or without manipulated DCAD had greater concentrations of mu-calpain and m-calpain mRNA, whereas the reverse was observed for calpastatin mRNA. Expression of mu calpain protein was increased relative to control by DCAD+25-OH D3. The amount of 25-OH D3 and manipulated DCAD administered to cull native Korean cows was insufficient to improve tenderness of beef by increasing muscle Ca concentration. However, DCAD+25-OH D3 induced greater expressions of mu-calpain protein as well as mRNA. PMID- 16699105 TI - Methane emissions from beef cattle: effects of fumaric acid, essential oil, and canola oil. AB - The objective of this study was to identify feed additives that reduce enteric methane emissions from cattle. We measured methane emissions, total tract digestibility (using chromic oxide), and ruminal fermentation (4 h after feeding) in growing beef cattle fed a diet supplemented with various additives. The experiment was designed as a replicated 4 x 4 Latin square with 21-d periods and was conducted using 16 Angus heifers (initial BW of 260 +/- 32 kg). Treatments were: control (no additive), fumaric acid (175 g/d) with sodium bicarbonate (75 g/d), essential oil and spice extract (1 g/d), or canola oil (4.6% of DMI). The basal diet consisted of 75% whole-crop barley silage, 19% steam-rolled barley, and 6% supplement (DM basis). Four large chambers (2 animals fed the same diet per chamber) were equipped to measure methane emissions for 3 d each period. Adding canola oil to the diet decreased (P = 0.009) total daily methane emissions by 32% and tended (P = 0.09) to decrease methane emissions as a percentage of gross energy intake by 21%. However, much of the reduction in methane emissions was due to decreased (P < 0.05) feed intake and lower (P < 0.05) total tract digestibility of DM and fiber. Digestibility of all nutrients was also lowered (P < 0.05) by feeding essential oil, but there were no effects on ruminal fermentation or methane emissions. In contrast, adding fumaric acid to the diet increased total VFA concentration (P = 0.03), increased propionate proportions (P = 0.01), and decreased the acetate:propionate ratio (P = 0.002), but there was no measurable effect on methane emissions. The study demonstrates that canola oil can be used to reduce methane losses from cattle, but animal performance may be compromised due to lower feed intake and decreased fiber digestibility. Essential oils had no effect on methane emissions, whereas fumaric acid caused potentially beneficial changes in ruminal fermentation but no measurable reductions in methane emissions. PMID- 16699106 TI - Effects of energy level on methionine utilization by growing steers. AB - We evaluated the effect of energy supplementation on Met use in growing steers. Six ruminally cannulated Holstein steers (228 +/- 8 kg of BW) were used in a 6 x 6 Latin square and fed 2.8 kg of DM/d of a diet based on soybean hulls. Treatments were abomasal infusion of 2 amounts of Met (0 or 3 g/d) and supplementation with 3 amounts of energy (0, 1.3, or 2.6 Mcal of GE/d) in a 2 x 3 factorial arrangement. The 1.3 Mcal/d treatment was supplied through ruminal infusion of 90 g/d of acetate, 90 g/d of propionate, and 30 g/d of butyrate, and abomasal infusion of 30 g/d of glucose and 30 g/d of fat. The 2.6 Mcal/d treatment supplied twice these amounts. All steers received basal infusions of 400 g/d of acetate into the rumen and a mixture (125 g/d) containing all essential AA except Met into the abomasum. No interactions between Met and energy levels were observed. Nitrogen balance was increased (P < 0.05) by Met supplementation from 23.6 to 27.8 g/d, indicating that protein deposition was limited by Met. Nitrogen retention increased linearly (P < 0.05) from 23.6 to 27.7 g/d with increased energy supply. Increased energy supply also linearly reduced (P < 0.05) urinary N excretion from 44.6 to 39.7 g/d and reduced plasma urea concentrations from 2.8 to 2.1 mM. Total tract apparent OM and NDF digestibilities were reduced linearly (P < 0.05) by energy supplementation, from 78.2 and 78.7% to 74.3 and 74.5%, respectively. Whole-body protein synthesis and degradation were not affected significantly by energy supplementation. Energy supplementation linearly increased (P < 0.05) serum IGF-I from 694 to 818 ng/mL and quadratically increased (P < 0.05) serum insulin (0.38, 0.47, and 0.42 ng/mL for 0, 1.3, and 2.6 Mcal/d, respectively). In growing steers, N retention was improved by energy supplementation, even when Met limited protein deposition, suggesting that energy supplementation affects the efficiency of AA use. PMID- 16699107 TI - Effects of energy source on methionine utilization by growing steers. AB - We evaluated the effects of different supplemental energy sources on Met use in growing steers. Ruminally cannulated Holstein steers were used in two 6 x 6 Latin squares, and data were pooled for analyses. In Exp. 1, steers (148 kg) were fed 2.3 kg of DM/d of a diet based on soybean hulls. Treatments (2 x 3 factorial) were abomasal infusion of 0 or 3 g of l-Met/d, and supplementation with no energy or with glucose (360 g/d) or fat (150 g/d) continuously infused into the abomasum. In Exp. 2, steers (190 kg) received 2.6 kg of dietary DM/d and were provided (2 x 3 factorial) with 0 or 3 g of l-Met/d, and with no supplemental energy or with acetate (385 g/d) or propionate (270 g/ d) continuously infused into the rumen. In both experiments, the energy sources supplied 1.3 Mcal of GE/d, and all steers received basal infusions of 400 g of acetate/d into the rumen and a mixture (125 g/d) of all essential AA except Met into the abomasum. Nitrogen balance (18.8 vs. 23.5 g/d; P < 0.01) and whole-body protein synthesis (2.1 vs. 2.3 kg/d; P < 0.07) were increased by Met supplementation, indicating that protein deposition was limited by Met. Supplemental energy reduced (P < 0.01) urinary N excretion and increased (P < 0.01) N retention without differences among energy sources. Increases in N retention in response to Met were numerically greater when energy was supplemented. Efficiency of supplemental Met use was 11% when no energy was supplemented but averaged 21% when 1.3 Mcal of GE/d was provided. Whole-body protein synthesis and degradation were not affected by energy supplementation. Serum insulin concentrations were increased by glucose and propionate supplementation. Serum IGF-I concentrations were increased by supplementation with Met or glucogenic sources of energy. In growing steers, N retention was increased by energy supplementation even though protein deposition was limited by Met, suggesting that energy supplementation improves the efficiency of AA use. These responses were independent of the source of energy. PMID- 16699108 TI - Effects of pig age at market weight and magnesium supplementation through drinking water on pork quality. AB - Thirty-two halothane-negative pigs (109 +/- 0.6 kg of BW) were used to determine the effect of pig age at marketing (and thus growth rate), and magnesium supplementation through drinking water, on pork quality. Two initial groups of 50 pigs that differed by 30 +/- 2 d of age were fed diets to meet or exceed nutrient requirements beginning at 28 kg of BW. Sixteen average, representative pigs were selected from each group to represent older, slow-growing pigs and younger, fast growing pigs. For the duration of the study, pigs were individually penned, provided 2.7 kg of feed (0.12% Mg) daily, and allowed free access to water. After 7 d of adjustment, pigs were blocked by sex and BW and allotted to 0 or 900 mg of supplemental Mg/L as MgSO4 in drinking water for 2 d before slaughter. All 32 pigs were then transported (110 km) to a commercial abattoir on the same day and slaughtered 2.5 h after arrival. Longissimus and semimembranosus (SM) chops were packaged and stored to simulate display storage for fluid loss and Minolta color determinations at 0, 2, 4, 6, and 8 d. Two remaining sections of the LM were vacuum-packaged and stored at 4 degrees C for 25 or 50 d. Fast- (younger) and slow- (older) growing pigs differed by 27 +/- 0.3 d of age (153 and 180 +/- 0.3 d; P < 0.001) at similar BW (108 and 110 +/- 0.6 kg of BW; P = 0.13). Supplementation of Mg tended to increase plasma Mg concentration (24.1 vs. 21.8 +/- 0.8 ppm; P = 0.06) but did not affect Mg concentration in LM or SM. Fluid loss of displayed LM or SM, and purge loss, color, and oxidation of vacuum packaged LM or SM were not affected by age or Mg (P > 0.10). Surface exudate of the SM from older pigs was lower than that of younger pigs (61 vs. 74 +/- 6 mg; P = 0.05) but was not different for the LM (P = 0.22). The LM from older pigs displayed for 4 and 8 d; P < 0.05) were less yellow (lower b*) than younger pigs. The SM from older pigs had lower lightness (L*) initially (47.9 vs. 49.5 +/- 0.4) and after 2 d (49.7 vs. 51.1 +/- 0.4), 6 d (52.1 vs. 53.7 +/- 0.4) and 8 d (54.5 vs. 55.9 +/- 0.4) of display storage. Younger pigs had greater oxidation of the LM than older pigs on d 8 of display (P < 0.01), and Mg decreased oxidation on d 8 within younger pigs (P < 0.05). Pork quality was improved in older pigs as indicated by less exudate, reduced yellowness of the LM, reduced paleness of the SM, and reduced oxidation of the LM. However, Mg supplementation through the water for 2 d did not affect pork quality of either older, slower growing pigs or younger, faster growing pigs. PMID- 16699109 TI - Comparison of cortisol, luteinizing hormone, and testosterone responses to a defined stressor in sexually inactive rams and sexually active female-oriented and male-oriented rams. AB - The objective of this study was to determine whether the effect of restraint stress on cortisol, LH, and testosterone varied among sexually inactive and sexually active female- and male-oriented rams, to allow differentiation among ram classes. Restraint stress or no stress was imposed on sexually inactive (n = 7) and sexually active female- (n = 17) and male-oriented (n = 6) rams in a 2 x 3 factorial arrangement. Rams were assigned to restraint or control within each classification. Rams were habituated to wearing halters and being tethered in separate pens, permitting visual, vocal, and olfactory contact with adjacent rams for 7 d before treatment. After 1 d of habituation, rams were fitted with jugular catheters that were checked twice daily for patency. For restraint stress, rams were laid on their side with their legs tied for 1 h. For no stress, rams were tethered with halters and leads, but their legs were not tied. On the treatment day, blood was collected at 30-min intervals for 3 h followed by 15-min intervals for 1 h before restraint, during 1-h restraint, and for 1 h after liberation from restraint. Then blood was collected at 30-min intervals for an additional 2 h. Blood was collected from controls at similar intervals. Control rams were isolated from stressed rams. Cortisol, LH, and testosterone were measured using RIA. Mixed model analyses with repeated measures were used on transformed data. Average prestress data were used as a covariate. Cortisol increased (P < 0.01) within 15 min after restraint and remained increased until 1.5 h after liberation from 1-h of restraint stress. In contrast, in controls cortisol remained unchanged at 5 ng/ mL. Cortisol did not differ over time among ram classes, and the treatment x ram class x time interaction was not significant. For LH and testosterone, the ram class x time interactions appeared to compromise the ability to identify differences in these hormones, indicating that they were not good endocrine candidates for methods of classifying rams. In conclusion, restraint stress increased cortisol in sexually inactive and sexually active female- and male-oriented rams alike, thus not providing a method to differentiate among ram classes. PMID- 16699110 TI - Influence of increased feeding frequency on behavior and integument lesions in growing-finishing restricted-fed pigs. AB - This study evaluated how feeding frequency affects behavior and the occurrence of skin lesions in growing-finishing pigs. One hundred eighty pigs (27 to 112 kg of BW) were reared in one environmentally controlled room (20 pens; 9 pigs/pen). Pigs in 10 pens were fed 3 times daily (reference group), whereas the others were fed 9 times daily (experimental group). Both groups received the same total amount of liquid feed. Rations were adjusted to the mean pen weights. Behavioral observations (scan sampling, as well as continuous focal pig observations) were made in wk 4, 10, and 14 of the growing-finishing period. After each observation, skin lesions were assessed individually for each pig. Pigs fed 9 times daily tended to lie laterally for less time (P = 0.083) and tended to be active (P = 0.054) during the day, especially in growing-finishing wk 4 (P = 0.007). With continuously observed focal pigs, no differences in time allocations for feeding were found between groups. During feeding in growing-finishing wk 4, focal pigs belonging to the experimental group displayed more aggressive actions (P = 0.019), tended to perform aggressive actions for a longer time (P = 0.076), and tended to be belly-nosed for a longer time (P = 0.083) compared with the reference group. In addition, in growing-finishing wk 14, pigs in the experimental group had greater scores for skin lesions (head, P = 0.001; belly, P < 0.001; caudal part, P < 0.001) and tended to be belly-nosed for a longer time (P = 0.084). In the case of pigs restricted-fed liquid feed, a greater frequency of feeding per day appears to be a condition that results in greater competitive feeding than with a lower feeding frequency. PMID- 16699111 TI - Effects of concentrated separator by-product (desugared molasses) on intake, ruminal fermentation, digestion, and microbial efficiency in beef steers fed grass hay. AB - Concentrated separator by-product (CSB) is produced when beet molasses goes through an industrial desugaring process. To investigate the nutritional value of CSB as a supplement for grass hay diets (12.5% CP; DM basis), 4 ruminally and duodenally cannulated beef steers (332 +/- 2.3 kg) were used in a 4 x 4 Latin square with a 2 x 2 factorial arrangement of treatments. Factors were intake level: ad libitum (AL) vs. restricted (RE; 1.25% of BW, DM basis) and dietary CSB addition (0 vs. 10%; DM basis). Experimental periods were 21 d in length, with the last 7 d used for collections. By design, intakes of both DM and OM (g/kg of BW) were greater (P < 0.01; 18.8 vs. 13.1 +/- 0.69 and 16.8 vs. 11.7 +/- 0.62, respectively) for animals consuming AL compared with RE diets. Main effect means for intake were not affected by CSB (P = 0.59). However, within AL-fed steers, CSB tended (P = 0.12) to improve DMI (6,018 vs. 6,585 +/- 185 g for 0 and 10% CSB, respectively). Feeding CSB resulted in similar total tract DM and OM digestion compared with controls (P = 0.50 and 0.87, respectively). There were no effects of CSB on apparent total tract NDF (P = 0.27) or ADF (P = 0.35) digestion; however, apparent N absorption increased (P = 0.10) with CSB addition. Total tract NDF, ADF, or N digestion coefficients were not different between AL- and RE-fed steers. Nitrogen intake (P = 0.02), total duodenal N flow (P = 0.02), and feed N escaping to the small intestine (P = 0.02) were increased with CSB addition. Microbial efficiency was unaffected by treatment (P = 0.17). Supplementation with CSB increased the rate of DM disappearance (P = 0.001; 4.9 vs. 6.9 +/- 0.33 %/h). Restricted intake increased the rate of in situ DM disappearance (P = 0.03; 6.4 vs. 5.3 +/- 0.33 %/h) compared with AL-fed steers. Ruminal DM fill was greater (P = 0.01) in AL compared with RE. Total VFA concentrations were greater (P = 0.04) for CSB compared with controls; however, ammonia concentrations were reduced (P = 0.03) with CSB addition. At different levels of dietary intake, supplementing medium-quality forage with 10% CSB increased N intake, small intestinal protein supply, and total ruminal VFA. PMID- 16699112 TI - Effect of processing flax in beef feedlot diets on performance, carcass characteristics, and trained sensory panel ratings. AB - To assess the effects of flax addition and flax processing on feedlot performance and carcass characteristics, 128 yearling beef heifers (360 +/- 14 kg of initial BW) were blocked by weight and assigned randomly to feedlot diets that included no flax (control), whole flax (WHL), rolled flax (RLD; 1,300 microm), or ground flax (GRD; 700 microm). Heifers were fed a growth diet (31% corn, 30% corn silage, 18% barley malt pellets, 14% alfalfa, 4% linseed meal, and 3% supplement; DM basis) for 56 d, after which they were adapted to a finishing diet (79% corn, 7% corn silage, 7% alfalfa, 4.75% linseed meal, and 2.25% supplement; DM basis). In WHL, RLD, and GRD, flax replaced all linseed meal and partially replaced corn at 8% of diet DM. All diets provided 0.5 mg of melengestrol acetate, 2,000 IU of vitamin E, and 232 mg of monensin per heifer daily. Cattle were slaughtered by block after 96, 97, and 124 (2 blocks) d on feed. At 24 h postmortem, carcass data were collected, and a portion of the loin was removed, vacuum-packaged, and aged for 14 d. After aging, 2 steaks were removed from each loin for Warner Bratzler shear force measurement, sensory panel evaluation, and fatty acid analysis (approximately 100 g of muscle was collected). Flax inclusion (WHL, RLD, and GRD vs. control) did not affect DMI (P = 0.79), fat thickness over the 12th rib (P = 0.32), or LM area (P = 0.23). Flax inclusion increased ADG (P = 0.006), G:F (P = 0.006), and USDA yield grade (P = 0.01). Flax processing (RLD and GRD vs. WHL) increased ADG (P = 0.05), G:F (P = 0.08), and apparent dietary NEm and NEg (P = 0.003). Muscle from heifers fed flax had greater phospholipid 18:3n-3 (P < 0.001), 20:5n-3 (P < 0.001), 22:5n-3 (P < 0.001), and 22:6n-3 (P = 0.02) fractions, and greater neutral lipid 18:3n-3 (P < 0.001). Feeding 8% flax to feedlot heifers increased gain and efficiency, and processing flax increased available energy and resulted in increased efficiency of gain. Feeding 8% flax also increased levels of n-3 fatty acids in fresh beef. PMID- 16699113 TI - A model for predicting feed intake of growing animals during exposure to pathogens. AB - A general model is proposed for predicting the effects of subclinical pathogen challenges of different doses and virulence on the relative feed intake (RFI) of animals. The RFI is defined as the feed intake (FI, kg/d) of the animal challenged by a pathogen divided by its FI in the same state had it not been challenged. Actual FI can be predicted from the RFI and the animal's state. The RFI was assumed to be affected only when animals were naive to a particular pathogen (i.e., had not previously experienced it) and when the challenge dose was above a predetermined threshold. The model is for the period from recognition of a pathogen through acquisition and subsequent expression of immunity. The way in which RFI changes with time is described by 5 main parameters and is based on data for RFI during different pathogen challenges of a range of hosts. Lag time (L, d) is the delay from a pathogen challenge until any effects on RFI are seen. Reduction time (R, d) describes the time it takes for the lowest value of RFI (lambda) to be achieved. The duration time (D, d) describes the time that lambda is maintained for, and rho (RFI/d) describes the rate of recovery of RFI until RFI = 1. There is no compensatory intake, and RFI is always < or = 1. The effects of host resistance on the values of the model parameters are proposed. Attempts were made to parameterize the model; when data were scarce, initial parameter values were derived on conceptual grounds. Predictions of the effects of pathogen dose, virulence, and host resistance are described and discussed. When comparing the responses in RFI for different genotypes, it is crucial to define the pathogen challenge (in terms of dose and virulence) and the degree of resistance of different hosts. Possible interactions between dose, virulence, and resistance were explored. Feed intake of healthy and challenged animals, at a time, may be different once the challenged animal has recovered (RFI = 1). The issue of reductions in FI during pathogen challenges is important for nutritionist and animal breeders. The large variation that has been observed for reductions in FI during pathogen challenges may be a viable point of selection. The points highlighted will aid selection strategies by quantifying the effects of pathogen dose and virulence, and time, on the FI of challenged animals. The proposed model may be integrated with other models of growth to predict animal performance during exposure to pathogens. PMID- 16699114 TI - Variations in meat pH of beef bulls in relation to conditions of transfer to slaughter and previous history of the animals. AB - Animals are subjected to various events that cause physical exhaustion and psychological stress during transfer to slaughter. This can lead to defective meat quality. Some animals may be better able to withstand the stress of transfer, depending on their previous experience of transport and on their finishing conditions (mixing, farmers' attitudes). The objective of this study was to assess the impact of 1) the conditions of transfer to slaughter (including duration of the journey, waiting time at lairage, etc.); and 2) the bulls' previous history (including experience in transport, mixing during finishing, and the farmers' attitudes) on the reactions of bulls to transfer and on their meat quality. We conducted a survey in commercial conditions. The history of the bulls and the facilities on the farms were noted; farmers were questioned on their attitudes; the bulls' reactions to loading into and unloading from the truck were observed; journey-related data were collected; and cortisol concentration at slaughter and the pH of the LM and the rectus abdominis were measured. Our study confirmed that certain physical factors associated with transport can increase stress and limit the decline of meat pH. These factors include the absence of loading facilities on the farm, transport on a warm day, or a short waiting time at lairage. Social aspects also played a role; the presence of bulls from the same finishing group limited stress and improved the pH decline. Events and management before transfer were also of importance; the farmer awareness of the sensitivity of bulls to humans or to feeding schedules but the absence of a positive attitude toward close contacts with bulls were all likely to limit stress or its consequences on meat pH. Although these results need to be confirmed in controlled experiments, they suggest that good management of beef bulls before and during transfer is essential to meat quality. PMID- 16699115 TI - Effect of long-term selection for increased leanness on meat and eating quality traits in Duroc swine. AB - A study was conducted to evaluate differences in meat and eating quality traits between purebred Duroc pigs sired by boars currently available and pigs sired by boars from the mid 1980s. Two lines were developed by randomly allocating littermate and half-sib pairs of females to matings by current time period (CTP) or old time period (OTP) boars. Matings by CTP boars were made using fresh semen, and matings by OTP boars were via frozen semen. All available barrows and randomly selected gilts were sent to a commercial abattoir and used for meat and eating quality evaluation. A total of 178 pigs from 23 CTP sires and 99 pigs from 15 OTP sires, across 2 replications and at a mean live weight of 109 kg, were slaughtered and analyzed. Chemical intramuscular fat percentage was determined by lab analysis of a slice from the LM at the 10th rib. Additional meat and eating quality traits measured on the LM were Minolta reflectance and Hunter L color (24 h); pH (24 h and 7 d); water-holding capacity; subjective visual scores for color, marbling, and firmness (48 h); Instron tenderness; cooking loss; and trained sensory panel evaluations (7 d). Time period differences were assessed by use of a mixed model that included fixed effects of sire time period, replication, sex, contemporary group, and the interaction of sex x time period. The random effect of dam and the random effect of sire nested within time period were also included. Loins from pigs sired by OTP boars had greater intramuscular fat (3.48 vs. 3.09%) and visual marbling scores (3.54 vs. 3.07), required less Instron force (5.31 vs. 5.98 kg) to compress, and had darker visual color scores (4.09 vs. 3.87) compared with loins from pigs sired by CTP boars (P < 0.05). No differences were observed between time periods for Minolta reflectance, Hunter L (24 h), water-holding capacity, pH (24 h and 7 d), or subjective firmness scores. Trained sensory evaluations revealed more pork flavor and less off-flavor (P < 0.05) for OTP-sired pigs; however, no differences in tenderness score, juiciness score, chewiness score, or cooking loss were found between lines. Long-term selection response in carcass composition has been at the expense of meat and eating quality traits. PMID- 16699116 TI - Manure composition of swine as affected by dietary protein and cellulose concentrations. AB - An experiment was conducted to investigate the effects of reducing dietary CP and increasing dietary cellulose concentrations on manure DM, C, N, S, VFA, indole, and phenol concentrations. Twenty-two pigs (105 kg initial BW) were fed diets containing either 14.5 or 12.0% CP, in combination with either 2.5 or 8.7% cellulose. Pigs were fed twice daily over the 56-d study, with feed intake averaging 2.74 kg/d. Feces and urine were collected after each feeding and added to the manure storage containers. Manure storage containers were designed to provide a similar unit area per animal as found in industry (7,393 cm2). Before sampling on d 56, the manure was gently stirred to obtain a representative sample for subsequent analyses. An interaction of dietary CP and cellulose was observed for manure acetic acid concentration, in that decreasing CP lowered acetic acid in pigs fed standard levels of cellulose but increased acetic acid in pigs fed greater levels of cellulose (P = 0.03). No other interactions were noted. Decreasing dietary CP reduced manure pH (P = 0.01), NH4 (P = 0.01), isovaleric acid (P = 0.06), phenol (P = 0.05), and 4-ethyl phenol (P = 0.02) concentrations. Increasing dietary cellulose decreased pH (P = 0.01) and NH4 (P = 0.07) concentration but increased manure C (P = 0.03), propionic acid (P = 0.01), butyric acid (P = 0.03), and cresol (P = 0.09) concentrations in the manure. Increasing dietary cellulose also increased manure DM (P = 0.11), N (P = 0.11), and C (P = 0.02) contents as a percentage of nutrient intake. Neither cellulose nor CP level of the diet affected manure S composition or output as a percentage of S intake. Headspace N2O concentration was increased by decreasing dietary CP (P = 0.03) or by increasing dietary cellulose (P = 0.05). Neither dietary CP nor cellulose affected headspace concentration of CH4. This study demonstrates that diets differing in CP and cellulose content can significantly impact manure composition and concentrations of VFA, phenol, and indole, and headspace concentrations of N(2)O, which may thereby affect the environmental impact of livestock production on soil, air, and water. PMID- 16699117 TI - Effects of plane of nutrition on in vitro fertilization and early embryonic development in sheep. AB - Nutrition has been shown to influence several reproductive functions, including hormone production, oocyte competence and fertilization, and early embryonic development. To determine the effects of maternal diet on in vitro fertilization (IVF) and early embryonic development, ewes (n = 18; 47.0 +/- 1.5 kg of initial BW) were divided into control and underfed (60% of control) nutritional planes for 8 wk before oocyte collection. Pelleted diets containing 2.4 Mcal of ME/kg and 13% CP (DM basis) were fed once daily. During the first 4-wk acclimation phase, control and underfed ewes were fed 1,000 and 600 g/d, respectively. From wk 4 to 8, control (adequate) ewes were fed to maintain BW and offered 720 g/d, whereas underfed ewes received 432 g/d (60% restricted). Synchronization of estrus was performed using progestagen sponges for 14 d. Follicular development was induced by twice daily injections of FSH on d 13 (5 units/injection) and 14 (4 units/injection) of the estrous cycle. Oocytes were collected from all visible follicles on d 15 of the estrous cycle. After IVF, the proportion of developing embryos was evaluated throughout an 8-d culture period. Under-nutrition decreased (P < 0.006) the rate of cleavage, number of blastocysts per ewe, and rate of blastocyst formation (from 79 to 64%; from 3.3 to 0.8; and from 31 to 8%, respectively). However, the number of visible follicles, total number of oocytes, number of healthy oocytes, percentage of healthy oocytes, number of cleaved oocytes, and morula formation per ewe were similar for control and underfed ewes. These data indicate that undernutrition of donor ewes, resulting in lower BW and BCS, has a negative effect on oocyte quality, which results in lower rates of cleavage and blastocyst formation. PMID- 16699118 TI - Effect of fescue toxicosis on hepatic gene expression in mice. AB - Fescue toxicosis affects wild and domestic animals grazing fescue pasture infected with the endophytic fungus Neotyphodium coenophialum. Signs of fescue toxicosis include increased core body temperature and respiration rate and decreased milk yield and reproductive performance. Laboratory mice also exhibit symptoms of fescue toxicosis, as indicated by reduced growth rate and reproductive performance. Mice were used to study the effects of fescue toxicosis on hepatic gene expression. Twenty-seven mice were randomly allocated to a diet containing either 50% endophyte-infected (E+; 6 ppm ergovaline) or endophyte-free (E-) fescue seed for 2 wk under thermoneutral conditions. Liver genes differentially expressed due to fescue toxicosis were identified using DNA microarray. A 2-stage ANOVA of microarray data identified 36 differentially expressed genes between mice fed E+ and E- diets. Another analysis method, significance analysis of microarray, identified 9 genes as differentially expressed between treatment groups, and some genes overlapped with genes identified by ANOVA. Hierarchical clustering of 36 genes identified by ANOVA clearly separated the mice by diet, with 100% confidence as computed by bootstrap analysis. Expression of 11 genes was verified using quantitative real-time PCR. The E+ diet resulted in downregulation of genes involved in the sex-steroid metabolism pathway and genes involved in cholesterol and lipid metabolism. Genes coding for ribosomes and protein synthesis were upregulated by the E+ diet. Genes identified in the present analysis indicate some of the mechanisms by which fescue toxicosis occurs in animals. PMID- 16699119 TI - Performance of beef heifers grazing stockpiled fescue as influenced by supplemental whole cottonseed. AB - The objectives of this study were to determine the composition of stockpiled fescue from December through February over 2 yr and to determine the performance of heifers grazing stockpiled fescue with or without supplemental whole cottonseed. In early December, 36 heifers (initial BW 277 +/- 0.70 kg, yr 1; 266 +/- 2.2 kg, yr 2; and initial BCS of 5.0 +/- 0.04) were assigned randomly to 6 groups. Each group was assigned randomly to a 2.4-ha tall fescue pasture (98% endophyte infected), which had received 76 kg of N/ha on September 1. Group was the experimental unit for all measures. Forage DM available during grazing (to ground level) averaged 3,913 and 5,370 kg/ha in yr 1 and 2, respectively. The pasture was strip-grazed for 83 d, with daily forage allocation. Three groups were fed whole cottonseed (0.90 kg of DM/heifer; 24.4% CP, DM basis) daily at 0.33% of BW, and a small amount of a corn-based concentrate (0.19 kg of DM/heifer) to assure complete cottonseed consumption. Nutritive value of forage (dry basis) was determined each week by sampling each pasture to the 5-cm target grazing height. Forage disappearance was estimated every 2 wk from pre- and postgraze forage mass. Forage CP was 16.8% in yr 1 and 12.6% in yr 2. In vitro true organic matter digestibility (IVTOMD) was 82.0 and 71.9%, and ADF was 25.9 and 30.7% in yr 1 and 2, respectively. Most indicators of forage quality declined slightly through the winter, although they recovered in late winter in yr 1. The proportion of fescue that was green declined (P < 0.05) from December (79% in yr 1 and 64% in yr 2) to February (62% in yr 1 and 52% in yr 2). Green tissue averaged 20.4 and 15.2% CP, 91 and 87% IVTOMD, and 22.1 and 23.3% ADF in yr 1 and 2, respectively. Brown tissue averaged 10.3 and 8.5% CP, 64 and 62% IVTOMD, and 35.7 and 37.4% ADF in yr 1 and 2, respectively. Shrunk ADG (0.46 vs. 0.56 kg/d in yr 1 and 0.23 vs. 46 kg/d in yr 2) and change in BCS (- 0.03 vs. 0.33 in yr 1 and 0.13 vs. 0.5 in yr 2) was greater (P < 0.05) for supplemented heifers. Supplemented heifers had greater serum urea nitrogen in yr 1 (9.5 vs. 10.5 mg/dL; P < 0.07) and yr 2 (7.2 vs. 8.6 mg/dL; P < 0.01). Forage disappearance was similar between supplemented and unsupplemented heifers (3.19 vs. 3.39 kg.heifer( 1).d(-1) in yr 1 and 4.14 vs. 4.17 kg.heifer(-1).d(-1) in yr 2, respectively). Heifers responded to supplementation, but performance was lower than expected based on forage nutrient content. PMID- 16699120 TI - Influence of stocking rate and steroidal implants on growth rate of steers grazing toxic tall fescue and subsequent physiological responses. AB - An 84-d grazing experiment was conducted in 2 growing seasons to evaluate interactions of stocking rate and steroidal implants with BW gain and symptoms of toxicosis in yearling steers grazing endemic endophyte-infected (E+) tall fescue (Festuca arundinacea Schreb.). A 4 x 2 factoral design was used to evaluate 4 stocking rates (3.0, 4.0, 5.0, and 6.0 steers/ ha) with or without steroidal implants (200 mg of progesterone + 20 mg of estradiol benzoate). Treatment combinations were randomly assigned to eight 1-ha pastures of E+ Kentucky-31 tall fescue (i.e., treatments were not replicated). Treatment effects were analyzed for ADG, total BW gain per hectare, forage availability, and hair coat ratings. At the conclusion of grazing in the second year (22 June), steers were placed on a bermudagrass [Cynodon dactylon (L.) Pers.] pasture, and rectal temperatures and serum prolactin concentrations were monitored for 10 d to assess carryover effects of stocking rate and steroidal implants on recovery from toxicosis related heat stress. Forage availability differed (P < 0.001) between years, but there were no year x treatment interactions (P > 0.10). There was an implant x stocking rate interaction (P < 0.05) on ADG. Differences between the slopes in the regression equations indicated that ADG responded to implantation when stocking rates were low, but the response diminished as stocking rate increased. Stocking rate did not influence (P = 0.89) postgraze rectal temperature, but the regression intercept for implanted steers was 0.4 degrees C greater (P < 0.05) than for nonimplanted steers, and the difference was consistent across the entire 10-d fescue-free grazing period. Concentrations of prolactin increased during the 10-d fescue-free grazing period, but trends differed due to an implantation x stocking rate interaction (P < 0.05). Results indicate that implantation with progesterone + estradiol benzoate increases ADG with lower stocking rates, but the effect diminishes with increased grazing intensity. Implantation with steroid hormones increased rectal temperatures, but during a fescue-free grazing period rectal temperatures and serum prolactins for implanted and nonimplanted steers returned to values indicative of a stable and healthy status in a 192- to 240-h (i.e., an 8- to 10-d) period. However, because the treatments used in this study were not replicated, these observations need to be confirmed with replicated studies. PMID- 16699121 TI - Perspective: a new ethic for health promotion: reflections on a philosophy of health education for the 21st century. AB - This article describes two models for thinking about the purposes of health education-a medical model and an education model--and traces how concerns about the validity of research have driven preference for the medical model. In the medical model, the purpose of health education is to develop effective interventions that will prevent people from adopting unhealthy behaviors. Here, health educators are expected to replicate the methods identified by researchers to effect targeted changes in health behavior. The article then describes an alternative way of thinking about the purposes of health education. In pursuing a philosophy of education, the purpose of research and practice would be to clarify basic social values and to strengthen one's faculty for making value judgments. Practitioners here use research results as a stimulus for dialogue about the role of good health habits in living the kind of life that community members find most valuable. PMID- 16699122 TI - An evaluation of the "TrEAT Yourself Well" restaurant nutrition campaign. AB - This study examined the effect of the "TrEAT Yourself Well" campaign on diners'menu choices using data from four restaurant chains in California. Within each chain, two locations in the greater San Diego area were selected as experimental sites and either one or two locations outside the greater San Diego area were selected as control sites. Various promotional activities, including in restaurant promotions, community events, and paid media advertising, were conducted in the experimental region to promote healthy menu entrees. The results show that the campaign was successful in reaching diners and had positive effects on their beliefs and attitudes toward healthy dining. The campaign directly increased the probability of a consumer purchasing a healthy menu item by 3.7% (p = .05). By improving consumer attitudes toward healthy menu items, the campaign indirectly increased purchases of these items by 4.4%. PMID- 16699123 TI - Results of a 3-year study of two methods of delivery of life skills training. AB - Model programs and standards for substance abuse prevention have been identified by a number of federal agencies. The study reported here assessed two methods of delivery of one such program, Life Skills Training (LST), implemented in nine rural disadvantaged school districts. The results indicate that neither standard LST nor an infused LST delivery method was found effective for the entire sample, although some encouraging results were found for the females in the study. This study, conducted by researchers independent of the LST program, is useful for school decision makers in determining what programs are most effective with which groups. It included all students with parental permission, controlling for prior use levels, unlike some previous LST studies. The results of the program, as implemented by regular classroom teachers, reflect many issues relevant to recruitment, training, implementation, adaptation, and institutionalization of prevention programming. PMID- 16699124 TI - Validity of a sun safety diary using UV monitors in middle school children. AB - This article describes a validity study conducted among middle school students comparing self-reported sun safety behaviors from a diary with readings from ultraviolet (UV) monitors worn on different body sites. The UV monitors are stickers with panels that turn increasingly darker shades of blue in the presence of increasing amounts of UV light. Three schools participated (N = 527). Statistically significant associations were found between self-reported behaviors and the UV monitor measurements, with Kendall's tau b's ranging from 0.400 to 0.823. Multiple regression was performed and indicator variables created separately for arm (R = .12) and leg (R = .49) coverage by protective clothing. The effect of time spent outdoors on the UV monitor measure was attenuated when the students reported having the corresponding body site covered with clothing. Self-reported shade was not a significant predictor of UV monitor change. Overall, the results provide evidence of validity of students' self-reports of time outdoors and protective clothing use. PMID- 16699125 TI - A systematic review of readability and comprehension instruments used for print and web-based cancer information. AB - Adequate functional literacy skills positively influence individuals' ability to take control of their health. Print and Web-based cancer information is often written at difficult reading levels. This systematic review evaluates readability instruments (FRE, F-K, Fog, SMOG, Fry) used to assess print and Web-based cancer information and word recognition and comprehension tests (Cloze, REALM, TOFHLA, WRAT) that measure people's health literacy. Articles on readability and comprehension instruments explicitly used for cancer information were assembled by searching MEDLINE and Psyc INFO from 1993 to 2003. In all, 23 studies were included; 16 on readability, 6 on comprehension, and 1 on readability and comprehension. Of the readability investigations, 14 focused on print materials, and 2 assessed Internet information. Comprehension and word recognition measures were not applied to Web-based information. None of the formulas were designed to determine the effects of visuals or design factors that could influence readability and comprehension of cancer education information. PMID- 16699126 TI - Nutrition education among low-income older adults: a randomized intervention trial in Congregate Nutrition sites. AB - Nutritional well-being among older adults is critical for maintaining health, increasing longevity, and decreasing the impact of chronic illness. However, few well-controlled studies have examined nutritional behavior change among low income older adults. A prospective, controlled, randomized design examined a five session nutrition education module delivered to limited-resource older adults (N = 703) in Congregate Nutrition sites by Cooperative Extension agents. Experimental group participants were significantly more likely than control group participants to increase multivitamin use, to increase calcium supplement use, to read labels of dietary supplements, to carry a supplement and/or medication list, and to discuss such use with their health care professional. The study addresses weaknesses in the literature by using a theoretically derived education component, implementing the intervention within a setting regularly used by low income older adults, employing randomized assignment to intervention and control conditions, and using hierarchical linear modeling to deal with "nested" data. PMID- 16699127 TI - Information desired and acquired by men with prostate cancer: data from ethnic focus groups. AB - Information and understanding are needed so that men with prostate cancer can effectively manage and cope with their disease. Although research has shown that cultural beliefs and practices influence the way people access health-related knowledge, little research has addressed how ethnicity affects the information desired and received among low-income men with prostate cancer. This research sought to (a) describe baseline knowledge at diagnosis, information subsequently received, information sources utilized, and knowledge believed to have been helpful and (b) explore differences in desired information among men of various ethnicities. Six ethnicity specific focus groups among men diagnosed with prostate cancer were conducted. With content analysis, themes emerging from each focus group were identified and compared. Results highlight disparities in the prostate cancer information desired and acquired by men of different ethnicities. PMID- 16699128 TI - Monitoring 14-3-3 protein interactions with a homogeneous fluorescence polarization assay. AB - The 14-3-3 proteins mediate phosphorylation-dependent protein-protein interactions. Through binding to numerous client proteins, 14-3-3 controls a wide range of physiological processes and has been implicated in a variety of diseases, including cancer and neurodegenerative disorders. To better understand the structure and function of 14-3-3 proteins and to develop small-molecule modulators of 14-3-3 proteins for physiological studies and potential therapeutic interventions, the authors have designed and optimized a highly sensitive fluorescence polarization (FP)-based 14-3-3 assay. Using the interaction of 14-3 3 with a fluorescently labeled phosphopeptide from Raf-1 as a model system, they have achieved a simple 1-step "mix-and-measure" method for analyzing 14-3-3 proteins. This is a solution-based, versatile method that can be used to monitor the binding of 14-3-3 with a variety of client proteins. The 14-3-3 FP assay is highly stable and has achieved a robust performance in a 384-well format with a demonstrated signal-to-noise ratio greater than 10 and a Z' factor greater than 0.7. Because of its simplicity and high sensitivity, this assay is generally applicable to studying 14-3-3/client-protein interactions and especially valuable for high-throughput screening of 14-3-3 modulators. PMID- 16699129 TI - A fluorescent microplate assay for exocytosis in alveolar type II cells. AB - The authors describe a simple, reliable, and quantitative assay to monitor exocytotic fusion of lamellar bodies (LBs) in adherent rat alveolar type II (AT II) cells. The assay is based on fluorescence measurements of LB-plasma membrane (PM) fusions modified for the use in multiwell culture plates to obtain a high sample throughput. In particular, it is based on the presence of a highly light absorbing dye in the cell supernatants to increase the specificity of fluorescence signals and to yield pseudo-confocal information from the cells. When the assay was tested with agonist-(ATP) and phorbolester-induced stimulation of LB-PM fusions, the authors found a good correlation with direct microscopic investigations based on single cell recordings. To further validate the assay, they used Curosurf at 10 mg/ml. However, it influenced neither the basal nor the ATP-stimulated rate of LB-PM fusions. This was corroborated by the fact that Curosurf had no effect on resting Ca (2+) levels nor the ATP induced Ca (2+) signals. The results cast new light on previous findings that surfactant phospholipids decrease the rate of secretion in AT II cells in a dose-dependent way. The authors conclude that the inhibitory effect exerted by phospholipids might be due to action on a later step in exocytosis, probably associated with exocytotic fusion pore expansion and content release out of fused vesicles. PMID- 16699130 TI - Multiparameter measurement of caspase 3 activation and apoptotic cell death in NT2 neuronal precursor cells using high-content analysis. AB - Caspase activation is a component of a number of neurodegenerative disorders, including stroke. In this study, the authors describe a multiplexed assay for caspase 3 activation, nuclear condensation, and cell viability in a neuronal precursor cell line Ntera-2, injured with staurosporine and etoposide. Using a high-content screening approach, cells were identified by staining with the nuclear stain Hoechst 33342; cell viability was measured by staining cells with YoPro-1, which is taken up by damaged cells but excluded from healthy cells; and caspase 3/7 activation was detected using the cell-permeable probe PhiPhi-Lux, which becomes fluorescent when cleaved by active caspase 3 or 7. These 3 dyes were detected simultaneously using a 4-band pass filter set on a Cellomics Array scan. The authors used peptide-fmk inhibitors selective for a variety of caspases, demonstrating that the injury is mediated primarily through caspase 3 or 7, although other caspases or related proteases may play a minor role. The general caspase inhibitor zVAD-fmkwas able to block cell death and caspase activation with the highest potency. The caspase 3 selective inhibitor DEVD fmkwas almost as potent as zVAD-fmk; other peptide caspase inhibitors displayed only modest inhibition of cell death. This assay was also used as a high-content screening tool for the evaluation of novel caspase 3 inhibitors for the potential treatment of degenerative disorders. PMID- 16699133 TI - Investigation of risk factors for clinical mastitis in British dairy herds with bulk milk somatic cell counts less than 150,000 cells/ml. AB - A sample of dairy farms in Great Britain with a monthly bulk milk somatic cell count of less than 150,000 cells/ml was enrolled into a 12-month prospective study. At the end of the study, a questionnaire on milking practice and other farm management practices was sent to the 482 farmers who had collected data on the occurrence of mastitis throughout the 12 months. The response rate was 93 per cent. The reported mean incidence of clinical mastitis was 36.9 cases per 100 cow years. Factors associated with an increase in the incidence of clinical mastitis were cleaning out the straw yard less frequently than every six weeks, more than 5 per cent of cows leaking milk outside the parlour, checking the foremilk, wearing gloves during milking, an average annual milk yield of more than 7000 litres per cow, dipping or spraying teats before milking and keeping milk with a high somatic cell count out of the bulk tank. Factors associated with a decrease in the incidence of clinical mastitis were using a cloth to dry the teats after washing them as part of premilking preparation, using calving boxes for less than 40 per cent of calvings, and using both cubicles and straw yards to house dry cows, as opposed to other housing. PMID- 16699134 TI - Effects of salinity and the use of shackle/leg sprays on the electrical conductivity of a commercial waterbath stunner for broilers. AB - The effect of salinity and temperature on the electrical conductivity of a waterbath stunner was assessed in a commercial broiler processing plant. During a break in production, the waterbath was emptied and filled with either cold water, cold water plus sodium chloride, warm water, or warm water plus sodium chloride. Different concentrations of sodium chloride in distilled water at different temperatures were prepared in the laboratory and their conductivities were compared with the results obtained in the processing plant. No significant increase in conductivity was achieved through an increase in water temperature in either the laboratory or the processing plant, but there was an increase in conductivity after the addition of sodium chloride. However, the increase in conductivity was lost after 20 to 30 minutes of birds passing through the waterbath. The effects of the use, the location and the water temperature of the shackle/leg sprays on electrical conductivity were also studied, but under the conditions of the experiment none of these factors increased the electrical conductivity. PMID- 16699135 TI - Prospective evaluation of postoperative pain in cats undergoing ovariohysterectomy by a midline or flank approach. AB - Twenty entire female cats were randomly assigned to two groups of 10; the cats in one group underwent ovariohysterectomy by a midline approach and the cats in the other group by a flank approach. Cats were assessed for signs of pain and scores were assigned pre- and postoperatively. There was a tendency for the cats neutered by a flank approach to be in more pain postoperatively (P=0.05). The final pain score for cats in either group was equal to or lower than their baseline score. PMID- 16699136 TI - Isolation of Brachyspira pilosicoli from weanling horses with chronic diarrhoea. PMID- 16699137 TI - Chronic eperythrozoonosis in an adult ewe. PMID- 16699138 TI - Cutaneous T cell lymphoma in a heifer seropositive for bovine leukosis virus. PMID- 16699139 TI - Nematode infestation with Heligmosomoides polygyrus in captive common dormice (Muscardinus avellanarius). PMID- 16699142 TI - Rare preputial anomaly and its surgical correction in a dog. PMID- 16699140 TI - TB policy and the badger culling trials. PMID- 16699143 TI - Natural and experimental West Nile virus infection in five raptor species. AB - We studied the effects of natural and/or experimental infections of West Nile virus (WNV) in five raptor species from July 2002 to March 2004, including American kestrels (Falco sparverius), golden eagles (Aquila chrysaetos), red tailed hawks (Buteo jamaicensis), barn owls (Tyto alba), and great horned owls (Bubo virginianus). Birds were infected per mosquito bite, per os, or percutaneously by needle. Many experimentally infected birds developed mosquito infectious levels of viremia (>10(5) WNV plaque forming units per ml serum) within 5 days postinoculation (DPI), and/ or shed virus per os or per cloaca. Infection of organs 15-27 days postinoculation was infrequently detected by virus isolation from spleen, kidney, skin, heart, brain, and eye in convalescent birds. Histopathologic findings varied among species and by method of infection. The most common histopathologic lesions were subacute myocarditis and encephalitis. Several birds had a more acute, severe disease condition represented by arteritis and associated with tissue degeneration and necrosis. This study demonstrates that raptor species vary in their response to WNV infection and that several modes of exposure (e.g., oral) may result in infection. Wildlife managers should recognize that, although many WNV infections are sublethal to raptors, subacute lesions could potentially reduce viability of populations. We recommend that raptor handlers consider raptors as a potential source of WNV contamination due to oral and cloacal shedding. PMID- 16699144 TI - Susceptibility of greater sage-grouse to experimental infection with West Nile virus. AB - Populations of greater sage-grouse (Centrocercus urophasianus) have declined 45 80% in North America since 1950. Although much of this decline has been attributed to habitat loss, recent field studies have indicated that West Nile virus (WNV) has had a significant negative impact on local populations of grouse. We confirm the susceptibility of greater sage-grouse to WNV infection in laboratory experimental studies. Grouse were challenged by subcutaneous injection of WNV (10(3.2) plaque-forming units [PFUs]). All grouse died within 6 days of infection. The Kaplan-Meier estimate for 50% survival was 4.5 days. Mean peak viremia for nonvaccinated birds was 10(6.4) PFUs/ml (+/-10(0.2) PFUs/ml, standard error of the mean [SEM]). Virus was shed cloacally and orally. Four of the five vaccinated grouse died, but survival time was increased (50% survival=9.5 days), with 1 grouse surviving to the end-point of the experiment (14 days) with no signs of illness. Mean peak viremia for the vaccinated birds was 10(2.3) PFUs/ml (+/-10(0.6) PFUs/ml, SEM). Two birds cleared the virus from their blood before death or euthanasia. These data emphasize the high susceptibility of greater sage grouse to infection with WNV. PMID- 16699145 TI - Pathology and preliminary characterization of a parapoxvirus isolated from a California sea lion (Zalophus californianus). AB - Cutaneous pox-like lesions are a common complication in the rehabilitation of pinnipeds. However, the exact identity, taxonomy, and host range of pinniped parapoxviruses remain unknown. During a poxvirus outbreak in May 2003 in California sea lions (Zalophus californianus) at a marine mammal rehabilitation facility, multiple raised, firm, 1-3-cm skin nodules from the head, neck, and thorax of one sea lion weanling pup that spontaneously died were collected. Histologically, the nodules were characterized by inflammation and necrosis of the dermis and epidermis, acanthosis, and ballooning degeneration of the stratum spinosum. Large, coalescing eosinophilic cytoplasmic inclusions were observed in the ballooned cells. A parapoxvirus (sea lion poxvirus 1, SLPV-1) was isolated on early passage California sea lion kidney cells inoculated with a tissue homogenate of a skin nodule. The morphology of the virions on electron microscopy was consistent with that of parapoxviruses. Partial sequencing of the genomic region encoding the putative major virion envelope antigen p42K confirmed the assignment of the sea lion poxvirus to the genus Parapoxvirus. Although SLPV-1 is most closely related to the poxvirus of harbor seals of the European North Sea, it is significantly different from orf virus, bovine papular stomatitis virus, pseudocowpox virus and the parapoxvirus of New Zealand red deer. PMID- 16699146 TI - Persistence of Pasteurella multocida in wetlands following avian cholera outbreaks. AB - Avian cholera, caused by Pasteurella multocida, affects waterbirds across North America and occurs worldwide among various avian species. Once an epizootic begins, contamination of the wetland environment likely facilitates the transmission of P. multocida to susceptible birds. To evaluate the ability of P. multocida serotype-1, the most common serotype associated with avian cholera in waterfowl in western and central North America, to persist in wetlands and to identify environmental factors associated with its persistence, we collected water and sediment samples from 23 wetlands during winters and springs of 1996 99. These samples were collected during avian cholera outbreaks and for up to 13 wk following initial sampling. We recovered P. multocida from six wetlands that were sampled following the initial outbreaks, but no P. multocida was isolated later than 7 wk after the initial outbreak sampling. We found no significant relationship between the probability of recovery of P. multocida during resampling and the abundance of the bacterium recovered during initial sampling, the substrate from which isolates were collected, isolate virulence, or water quality conditions previously suggested to be related to the abundance or survival of P. multocida. Our results indicate that wetlands are unlikely to serve as a long-term reservoir for P. multocida because the bacterium does not persist in wetlands for long time periods following avian cholera outbreaks. PMID- 16699147 TI - Lesions associated with a novel Mycoplasma sp. in California sea lions (Zalophus californianus) undergoing rehabilitation. AB - From July 1999 to November 2001, Mycoplasma sp. was cultured from lesions in 16 California sea lions (Zalophus californianus) undergoing rehabilitation. The Mycoplasma sp. was the likely cause of death of four animals in which it was associated with either pneumonia or polyarthritis. The most common lesion associated with this bacterium was subdermal abscessation, found in 12 animals. Other lesions included intramuscular abscesses, septic arthritis, and lymphadenopathy. Infection was associated with a leukocytosis and left shift in 12 animals. Animals with abscesses improved clinically after surgical lancing, irrigation, and systemic antibiotic therapy. The mycoplasma isolates had a consistent 16S rRNA sequence dissimilar from other Mycoplasma spp. and represent a novel species, Mycoplasma zalophi proposed sp. nov. PMID- 16699148 TI - Prevalence of shiga toxin-producing Escherichia coli and Salmonella enterica in rock pigeons captured in Fort Collins, Colorado. AB - The potential role of rock pigeons (Columba livia) in the epidemiology of shiga toxin-producing Escherichia coli (STEC) and Salmonella enterica is unclear. Our objective was to determine the prevalence of STEC and S. enterica in pigeons at urban and dairy settings as a function of season. Prevalence of STEC and S. enterica was estimated by bacteriologic culture of cloacal swabs collected from pigeons trapped at urban and dairy locations in and around Fort Collins, Colorado from January to November 2003. Presumptive E. coli isolates were tested for the presence of virulence genes SLT-1, SLT-2, eae, hlyA, K1, CNF-1, CNF-2, and LT using polymerase chain reaction. Shiga toxins were not isolated from any of 406 samples from pigeons, but virulence genes typically associated with disease in humans were identified in isolates from 7.9% (95% CI: 5.5% to 10.9%) of captured pigeons. S. enterica were detected in 3.2% of 277 samples from pigeons, with all positive samples originating from dairy locations (nine of 106 [8.5%]; 95% CI: 4.0-15.5%). The results suggest that although pigeons may acquire S. enterica from cattle and play a role in recirculation and persistence of the microorganism at dairies, pigeons are not important carriers of STEC. PMID- 16699149 TI - Necropsy findings in American alligator late-stage embryos and hatchlings from northcentral Florida lakes contaminated with organochlorine pesticides. AB - Increased American alligator (Alligator mississippiensis) embryo and neonatal mortality has been reported from several northcentral Florida lakes contaminated with old-use organochlorine pesticides (OCPs). However, a clear relationship among these contaminants and egg viability has not been established, suggesting the involvement of additional factors in these mortalities. Thus, the main objective of this study was to determine the ultimate cause of mortality of American alligator late-stage embryos and hatchlings through the conduction of detailed pathological examinations, and to evaluate better the role of OCPs in these mortalities. Between 2000 and 2001, 236 dead alligators were necropsied at or near hatching (after approximately 65 days of artificial incubation and up to 1 mo of age posthatch). Dead animals were collected from 18 clutches ranging in viability from 0% to 95%. Total OCP concentrations in yolk ranged from approximately 100 to 52,000 microg/kg, wet weight. The most common gross findings were generalized edema (34%) and organ hyperemia (29%), followed by severe emaciation (14%) and gross deformities (3%). Histopathologic examination revealed lesions in 35% of the animals, with over half of the cases being pneumonia, pulmonary edema, and atelectasis. Within and across clutches, dead embryos and hatchlings compared with their live cohorts were significantly smaller and lighter. Although alterations in growth and development were not related to yolk OCPs, there was an increase in prevalence of histologic lesions in clutches with high OCPs. Overall, these results indicate that general growth retardation and respiratory abnormalities were a major contributing factor in observed mortalities and that contaminants may increase the susceptibility of animals to developing certain pathologic conditions. PMID- 16699150 TI - A plague epizootic in the black-tailed prairie dog (Cynomys ludovicianus). AB - Plague is the primary cause for the rangewide decline in prairie dog (Cynomys spp.) distribution and abundance, yet our knowledge of plague dynamics in prairie dog populations is limited. Our understanding of the effects of plague on the most widespread species, the black-tailed prairie dog (C. ludovicianus), is particularly weak. During a study on the population biology of black-tailed prairie dogs in Wyoming, USA, plague was detected in a colony under intensive monitoring, providing a unique opportunity to quantify various consequences of plague. The epizootic reduced juvenile abundance by 96% and adult abundance by 95%. Of the survivors, eight of nine adults and one of eight juveniles developed antibodies to Yersinia pestis. Demographic groups appeared equally susceptible to infection, and age structure was unaffected. Survivors occupied three small coteries and exhibited improved body condition, but increased flea infestation compared to a neighboring, uninfected colony. Black-tailed prairie dogs are capable of surviving a plague epizootic and reorganizing into apparently functional coteries. Surviving prairie dogs may be critical in the repopulation of plague-decimated colonies and, ultimately, the evolution of plague resistance. PMID- 16699151 TI - Multi-species patterns of avian cholera mortality in Nebraska's Rainwater Basin. AB - Nebraska's Rainwater Basin (RWB) is a key spring migration area for millions of waterfowl and other avian species. Avian cholera has been endemic in the RWB since the 1970s and in some years tens of thousands of waterfowl have died from the disease. We evaluated patterns of avian cholera mortality in waterfowl species using the RWB during the last quarter of the 20th century. Mortality patterns changed between the years before (1976-1988) and coincident with (1989 1999) the dramatic increases in lesser snow goose abundance and mortality. Lesser snow geese (Chen caerulescens caerulescens) have commonly been associated with mortality events in the RWB and are known to carry virulent strains of Pasteurella multocida, the agent causing avian cholera. Lesser snow geese appeared to be the species most affected by avian cholera during 1989-1999; however, mortality in several other waterfowl species was positively correlated with lesser snow goose mortality. Coincident with increased lesser snow goose mortality, spring avian cholera outbreaks were detected earlier and ended earlier compared to 1976-1988. Dense concentrations of lesser snow geese may facilitate intraspecific disease transmission through bird-to-bird contact and wetland contamination. Rates of interspecific avian cholera transmission within the waterfowl community, however, are difficult to determine. PMID- 16699152 TI - Wild bird mortality and West Nile virus surveillance: biases associated with detection, reporting, and carcass persistence. AB - Surveillance targeting dead wild birds, in particular American crows (Corvus brachyrhynchos), plays a critical role in West Nile virus (WNV) surveillance in the United States. Using crow decoy surrogates, detection and reporting of crow carcasses within urban and rural environments of DeKalb County, Georgia were assessed for potential biases that might occur in the county's WNV surveillance program. In each of two replicated trials, during July and September 2003, 400 decoys were labeled with reporting instructions and distributed along randomly chosen routes throughout designated urban and rural areas within DeKalb County. Information-theoretic methods were used to compare alternative models incorporating the effects of area and trial on probabilities of detection and reporting. The model with the best empirical support included the effects of area on both detection and reporting of decoys. The proportion of decoys detected in the urban area (0.605, SE=0.024) was approximately twice that of the rural area (0.293, SE=0.023), and the proportion of decoys reported in the urban area (0.273, SE=0.023) was approximately three times that of the rural area (0.103, SE=0.028). These results suggest that human density and associated factors can substantially influence dead crow detection and reporting and, thus, the perceived distribution of WNV. In a second and separate study, the persistence and fate of American crow and house sparrow (Passer domesticus) carcasses were assessed in urban and rural environments in Athens-Clarke, Madison, and Oconee counties, Georgia. Two replicated trials using 96 carcasses of each species were conducted during July and September 2004. For a portion of the carcasses, motion sensitive cameras were used to monitor scavenging species visits. Most carcasses (82%) disappeared or were decayed by the end of the 6-day study. Carcass persistence averaged 1.6 days in rural areas and 2.1 days in urban areas. We analyzed carcass persistence rates using a known-fate model framework in program MARK. Model selection based on Akaike's Information Criteria (AIC) indicated that the best model explaining carcass persistence rates included species and number of days of exposure; however, the model including area and number of days of exposure received approximately equal support. Model-averaged carcass persistence rates were higher for urban areas and for crow carcasses. Six mammalian and one avian species were documented scavenging upon carcasses. Dead wild birds could represent potential sources of oral WNV exposure to these scavenging species. Species composition of the scavenger assemblage was similar in urban and rural areas but "scavenging pressure" was greater in rural areas. PMID- 16699153 TI - Hematology, plasma biochemistry, and antibodies to select viruses in wild-caught Eastern massasauga rattlesnakes (Sistrurus catenatus catenatus) from Illinois. AB - During the 2004 field season, blood was collected from Eastern massasauga rattlesnakes (Sistrurus catenatus catenatus) in the Carlyle Lake (Carlyle, Illinois, USA) and Allerton Park (Monticello, Illinois, USA) populations to derive baseline complete blood count and plasma biochemistry data and to assess the prevalence of antibodies to West Nile virus (WNV) and ophidian paramyxovirus (OPMV). Massasaugas were located for sampling through visual encounter surveys. Body weight, snout-vent length, total protein, globulins, sodium, and potassium were normally distributed among the survey population. Aspartate aminotransferase, creatine kinase, albumin, calcium, uric acid, white blood cell count, heterophils, lymphocytes, monocytes, eosinophils, and basophils were non normally distributed within these animals. Female snakes had significantly shorter tail lengths; lower blood glucose, packed cell volumes, and absolute azurophil counts; and higher plasma calcium and phosphorus concentrations than did males. None of the snakes tested (n=21) were seropositive for WNV, whereas all (n=20) were seropositive for OPMV. PMID- 16699154 TI - Hematology and serum chemistry of harp (Phoca groenlandica) and hooded seals (Cystophora cristata) during the breeding season, in the Gulf of St. Lawrence, Canada. AB - Standard hematologic and serum chemistry parameters were determined from 28 harp seals (Phoca groenlandica) and 20 hooded seals (Cystophora cristata) sampled from 6 March 2001 to 13 March 2001 during the breeding season. Whole blood was collected immediately postmortem from harp seal mother-pup pairs and from six hooded seal pups, and from live-captured adult hooded seals and three hooded seal pups; blood was analyzed within 24 hr at a local human hospital. A certified veterinary laboratory validated subsamples of whole blood and analyzed all serum chemistry parameters. Significant interlaboratory differences in mean values of packed cell volume (PCV) and mean cell volume (MCV) were found. Significant differences were found between samples from the five seal groups (adult male hooded seals, lactating female hooded seals, unweaned hooded seal pups; lactating female harp seals, and unweaned harp seal pups) for hematology and most serum chemistry parameters. In general, age-class influenced mean values of PCV, hemoglobin (HB), red blood cell (RBC) counts, MCV, mean cell hemoglobin (MCH), mean cell hemoglobin concentration (MCHC), and nucleated red blood cell (NRBC) counts per 100 leucocytes, but most age-related variations were species specific. Harp seal pups had significantly lower mean values of HB, PCV, MCH, and MCHC than did other seal groups, and significantly lower mean RBC counts than did hooded seal pups. Mean NRBC counts per 100 leukocytes were more than three times higher in harp seal pups than in hooded seal pups, but this difference was not statistically significant. Mean MCV were significantly lower in harp and hooded seal pups compared to those of adult harp and hooded seals. Differences in hemograms between pup species were likely because of the precocious development of hooded seal pups, which are weaned within 4 days, compared to 12 days for harp seal pups. Among adult seal groups, male hooded seals had significantly higher mean values of PCV and HB than did female harp and hooded seals, and significantly higher mean RBC counts than did adult female hooded seals. Among adult females, mean values of MCH and MCHC were statistically higher in hooded seals than in harp seals. Adult female harp and hooded seals did not differ significantly in other RBC parameters and mean leukocyte counts. Mean values of glucose, blood urea nitrogen, total bilirubin, alanine aminotransferase (ALT), alkaline phosphatase (ALP), total protein, and albumin showed species-specific variations between adults and pups. Except for ALP, few significant differences in mean enzyme activities of aspartate aminotransferase (AST), ALT, creatine kinase and gamma-glutamyltransferase were found between seal groups. Mean concentrations of electrolytes (calcium, phosphorus, sodium, potassium, chloride, magnesium, and total carbon dioxide) varied with age class, but variations in potassium and magnesium were species specific. Harp seal pups had significantly higher mean phosphorus and potassium levels compared to other seal groups. PMID- 16699155 TI - Hematology, plasma chemistry, and serology of the flightless cormorant (Phalacrocorax harrisi) in the Galapagos Islands, Ecuador. AB - The flightless cormorant (Phalacrocorax harrisi) is an endemic species of the Galapagos Islands, Ecuador. Health studies of the species have not previously been conducted. In August 2003, baseline samples were collected from flightless cormorant colonies on the islands of Isabela and Fernandina. Seventy-six birds, from nestlings to adults, were evaluated. Genetic sexing of 70 cormorants revealed 37 females and 33 males. Hematology assessment consisted of packed cell volume (n=19), leukograms (n=69), and blood smear evaluation (n=69). Microscopic evaluation of blood smears revealed microfilaria in 33% (23/69) of the cormorants. Plasma chemistries were performed on 46 cormorants. There was no significant difference in chemistry values or complete blood counts between male and female cormorants or between age groups. Based on a serologic survey to assess exposure to avian pathogens, birds (n=69) were seronegative for West Nile virus, avian paramyxovirus type 1 (Newcastle disease virus), avian paramyxovirus types 2 and 3, avian influenza, infectious bursal disease, infectious bronchitis, Marek's disease (herpes), reovirus, avian encephalomyelitis, and avian adenovirus type 2. Antibodies to avian adenovirus type 1 and Chlamydophila psittaci were found in 31% (21/68) and 11% (7/65) of flightless cormorants respectively. Chlamydophila psittaci was detected via polymerase chain reaction in 6% (2/33) of the cormorants. The overall negative serologic findings of this research suggest that the flightless cormorant is an immunologically naive species, which may have a reduced capacity to cope with the introduction of novel pathogens. PMID- 16699156 TI - A novel gammaherpesvirus associated with genital lesions in a Blainville's beaked whale (Mesoplodon densirostris). AB - An adult male Blainville's beaked whale (Mesoplodon densirostris) was found stranded on the Atlantic coast of the USA on 28 January 2004. Necropsy revealed a focal papilloma-like penile lesion, the cells from which revealed single 4-6 microm basophilic intranuclear inclusions. Total DNA extracted from lesion material was tested using a pan-herpes-virus PCR assay that targets the DNA polymerase gene and found to be positive. When the amplified DNA fragment was cloned, sequenced, and compared to GenBank-deposited herpesvirus DNA polymerase sequences, the detected virus was determined to be a distinct member of the Gammaherpesvirinae subfamily of herpesviruses. This new virus, tentatively named Ziphiid herpesvirus type 1, was associated with but not determined to be the cause of genital disease in the Blainville's beaked whale. PMID- 16699157 TI - Papillomatosis in a European bison. AB - Five European bison (Bison bonasus) from three European zoos were shipped to the Bukovske Vrchy Hills (Slovakia) in June 2004 and kept together in an acclimatization enclosure. The European bison were released into the wild in December 2004. At that time, papillomas were found at the medial canthus of the left eye of a 12-yr-old female bison. Cutaneous papillomatosis was confirmed histologically. Negative stain transmission electron microscopic examination revealed papillomavirus in the papillomas, and papillomavirus DNA also was detected using the polymerase chain reaction with FAP59 and FAP64 primers. The amplified 413 bp DNA sequence was identical to that of BAPV2 bovine papillomavirus. This paper is the first report of papillomatosis in European bison. PMID- 16699158 TI - Exposure of Mongolian gazelles (Procapra gutturosa) to foot and mouth disease virus. AB - Foot and mouth disease is a highly contagious acute viral disease that affects most ruminant and porcine species. During 2001, 33 serum samples were collected from Mongolian gazelles (Procapra gutturosa) in the Eastern Steppe of Mongolia. Samples were tested for antibodies to seven subtypes of foot-and-mouth-disease virus (FMDV). Antibodies were detected in 67% of the animals, and serologic results indicated exposure to FMDV-O. This virus was present in domestic animal populations in Mongolia from 2000 to 2002, and it is likely that the antibodies to FMDV detected in these gazelles resulted from spillover of virus from domestic animal sources. PMID- 16699159 TI - Determining prevalence of bluetongue and epizootic hemorrhagic disease viruses in mule deer in Arizona (USA) using whole blood dried on paper strips compared to serum analyses. AB - We investigated the feasibility of using whole blood dried on paper strips as a means to collect antibody prevalence data for the epizootic hemorrhagic disease viruses (EHDV) and bluetongue viruses (BTV) from hunter-harvested male mule deer (Odocoileus hemionus) in October 2002 from Arizona, USA. We compared antibody prevalence estimates in mule deer from paired paper strip and serum samples. Prevalence data obtained from elution of dried blood on paper strips proved to be consistent with results from serum in 94% of the samples tested. The paper strip method allows easy collection of blood from dead animals, with a smaller amount of blood being needed for analyses. Also, samples do not need to be refrigerated before analyses. We also used serum samples to determine hemorrhagic disease (HD) serotype exposure status of mule deer harvested from 4 distinct areas in Arizona. Antibodies to BTV and EHDV were identified in 3 of the 4 areas, with positive results to EHDV-1, EHDV-2, BTV-10, and BTV-11 being most common. Many animals did not have antibodies against the BTV serotypes. Exposure varied geographically and potentially with elevation. Hemorrhagic disease viruses commonly infect Arizona mule deer, except on the Kaibab Plateau in northern Arizona. PMID- 16699160 TI - No evidence of persistent Yersina pestis infection at prairie dog colonies in north-central Montana. AB - Sylvatic plague is a flea-borne zoonotic disease caused by the bacterium Yersinia pestis, which can cause extensive mortality among prairie dogs (Cynomys) in western North America. It is unclear whether the plague organism persists locally among resistant host species or elsewhere following epizootics. From June to August 2002 and 2003 we collected blood and flea samples from small mammals at prairie dog colonies with a history of plague, at prairie dog colonies with no history of plague, and from off-colony sites where plague history was unknown. Blood was screened for antibody to Y. pestis by means of enzyme-linked immunosorbent assay or passive hemagglutination assay and fleas were screened for Y. pestis DNA by polymerase chain reaction. All material was negative for Y. pestis including 156 blood samples and 553 fleas from colonies with a known history of plague. This and other studies provide evidence that Y. pestis may not persist at prairie dog colonies following an epizootic. PMID- 16699161 TI - A paretic condition in an Anaplasma phagocytophilum infected roe deer calf. AB - This paper describes a case of Anaplasma phagocytophilum infection in a roe deer (Capreolus capreolus) calf in Norway. The calf was found deserted, paretic, and heavily infested with Ixodes ricinus ticks. It was euthanized and investigated postmortem. Anaplasma phagocytophilum was detected in several tissues by polymerase chain reaction (PCR) and 16S rRNA sequence analyses. Analyses for Borrelia burgdorferi sensu lato and tick-borne encephalitis (TBE) virus infections were negative. This is the first report of a possible paretic condition in A. phagocytophilum infected roe deer. PMID- 16699162 TI - Cryptococcus gattii in wildlife of Vancouver Island, British Columbia, Canada. AB - Although Cryptococcus gattii has emerged as an important pathogen of humans and domestic animals on Vancouver Island, Canada since 1999; its distribution in regional wildlife species is largely unknown. Opportunistic sampling methods were employed to obtain nasal swabs for fungal culture from wild mammal species residing within the coastal Douglas fir biogeoclimatic zone on the southeast coast of the island. Samples were collected from 91 animals representing 14 species. Cryptococcus gattii was isolated from the nasal swabs of two eastern gray squirrels (Sciurus carolinensis) trapped in Duncan, British Columbia. The relative proportion of nasal colonization in wild mammal species is consistent with findings in domestic animals, suggesting that animals may be good indicators of environmental organisms. PMID- 16699163 TI - Antibodies to Toxoplasma gondii in Eurasian badgers. AB - Antibodies to Toxoplasma gondii were detected in samples collected from 90 live trapped adult Eurasian badgers (Meles meles) sampled at three sites (two agricultural and one woodland) in southern England. Serum was tested using a qualitative latex agglutination test procedure and 63 of 90 (70%) badgers tested positive for T. gondii antibodies. Antibody prevalence varied between the sites; 67% and 77% of badgers from agricultural sites and 39% from a nonagricultural site tested positive. PMID- 16699164 TI - Serological survey of Toxoplasma gondii infection in free-ranging Eurasian lynx (Lynx lynx) from Sweden. AB - To investigate the prevalence of Toxoplasma gondii infection in free-ranging Eurasian lynx (Lynx lynx) in Sweden, serosanguinous fluids and feces were collected from 207 carcasses of lynx killed or found dead from 1996 to 1998. Sera were tested for antibodies against T. gondii by the direct agglutination test, and 156 (75.4%) of the sera tested positive at antibody titers>or=40. Antibody prevalence was significantly lower in lynx originating from the northern parts of Sweden than in lynx from the more southern regions that are more densely populated by humans. Age-related differences also were found, with a significantly lower prevalence (55%) in juvenile (<1-yr-old) than in subadult and adult animals (82%). There was no significant difference in seroprevalence between males and females. Oocysts typical of T. gondii were not detected in any of the fecal samples. PMID- 16699165 TI - Seroprevalence of antibodies to Toxoplasma gondii in the Pennsylvania bobcat (Lynx rufus rufus). AB - From 2000 to 2002 bobcat blood samples were collected, in association with the Pennsylvania Game Commission, during the recently reactivated bobcat hunting and trapping season. Sex, age, and county/township data were recorded for each animal. Blood was tested for antibodies to Toxoplasma gondii using the modified agglutination test. In the 2-yr study, 131 bobcat samples were collected in 14 Pennsylvania counties and 109 (83%) of these had antibodies to T. gondii (titer>or=25). A two-way Chi-Square test (95% confidence interval) yielded no significance differences in antibody prevalence between males (83%) and females (88%) or adults (83%) and juveniles (77%). All 14 counties had at least one bobcat with antibodies to T. gondii. PMID- 16699166 TI - Stearylated octaarginine and artificial virus-like particles for transfection of siRNA into primary rat neurons. AB - RNA interference (RNAi) provides a powerful experimental tool for sequence specific gene silencing, allowing efficient analysis of gene function in a multitude of cell types. However, application of RNAi in primary mammalian neurons has been limited by low-transfection efficiency and considerable toxicity of conventional transfection methods. In this study, we evaluated a peptide mediated and a polymer/lipid-based cellular delivery method for siRNA into rat primary neurons and compared the results with a commonly used liposomal transfection reagent. Stearylated octaarginine (Stearyl-R8) was used as polypeptide and artificial virus-like particles (AVPs) were used as a combined liposomal-polymeric vector, since both reagents have been previously shown to successfully transfect DNA into cell lines. Stearyl-R8 and AVPs both promoted siRNA transfection into primary hippocampal neurons via the endosomal pathway. SiRNA-mediated gene silencing could be effectively induced in primary neuron cultures. In comparison with the commonly used cationic liposome transfection agent, both novel reagents were less detrimental to cell metabolic activity. We conclude that these novel transfection methods yield performances comparable to cationic liposome-mediated transfection for siRNA, while being less cytotoxic in primary neurons. Stearyl-R8 and AVPs may therefore represent novel and more cost efficient alternatives to conventional siRNA-transfection reagents. PMID- 16699167 TI - Characterization of GE82832, a peptide inhibitor of translocation interacting with bacterial 30S ribosomal subunits. AB - GE82832, a secondary metabolite produced by Streptosporangium cinnabarinum (strain GE82832), has been identified as a translational inhibitor by in vitro screening of a library of natural products. Secondary functional tests specific for individual steps of the translational pathway demonstrated that translocation is the specific target of GE82832. Chemical probing in situ demonstrated that this antibiotic protects bases A1324 and A1333 and exposes C1336 of 16S rRNA, thereby indicating that its binding site is located on the head of the 30S ribosomal subunit. The ribosomal location of GE82832, near ribosomal protein S13 and G1338, two elements of the small subunit that are part of or close to the B1a intrasubunit bridge, suggests that translocation inhibition results from an altered dynamics of 30S-50S ribosomal subunit interaction. PMID- 16699168 TI - Modulators of cigarette smoke-induced pulmonary emphysema in A/J mice. AB - Mice develop pulmonary emphysema after chronic exposure to cigarette smoke (CS). In this study, the influence of gender, exposure duration, and concentration of CS on emphysema, pulmonary function, inflammation, markers of toxicity, and matrix metalloproteinase (MMP) activity was examined in A/J mice. Mice were exposed to CS at either 100 or 250 mg total particulate material/m(3) (CS-100 or CS-250, respectively) for 10, 16, or 22 weeks. Evidence of emphysema was first seen in female mice after 10 weeks of exposure to CS-250, while male mice did not develop emphysema until 16 weeks. Female mice exposed to CS-100 did not have emphysema until 16 weeks, suggesting that disease development depends on the concentration and duration of exposure. Airflow obstruction and increased pulmonary compliance were observed in mice exposed to CS-250 for 22 weeks. Decreased elasticity was likely the major contributor to airflow obstruction because substantial remodeling of the conducting airways, beyond mild mucous cell hyperplasia, was lacking. Exposure to CS increased the number of macrophages, neutrophils, lymphocytes (B cells and activated CD4- and CD8-positive T cells), and activity of MMP-2 and -9 in the bronchoalveolar lavage fluid (BALF). Treatment with antioxidants N-acetylcysteine or epigallocatechin gallate (EGCG) did not decrease emphysema severity, but EGCG slightly decreased BALF inflammatory cell numbers and lactate dehydrogenase activity. Inflammation and emphysema persisted after a 17-week recovery period following exposure to CS-250 for 22 weeks. The similarities of this model to the human disease make it promising for studying disease pathogenesis and assessing new therapeutic interventions. PMID- 16699169 TI - Heterotachy processes in rhodophyte-derived secondhand plastid genes: Implications for addressing the origin and evolution of dinoflagellate plastids. AB - Serial transfer of plastids from one eukaryotic host to another is the key process involved in evolution of secondhand plastids. Such transfers drastically change the environment of the plastids and hence the selection regimes, presumably leading to changes over time in the characteristics of plastid gene evolution and to misleading phylogenetic inferences. About half of the dinoflagellate protists species are photosynthetic and unique in harboring a diversity of plastids acquired from a wide range of eukaryotic algae. They are therefore ideal for studying evolutionary processes of plastids gained through secondary and tertiary endosymbioses. In the light of these processes, we have evaluated the origin of 2 types of dinoflagellate plastids, containing the peridinin or 19'-hexanoyloxyfucoxanthin (19'-HNOF) pigments, by inferring the phylogeny using "covarion" evolutionary models allowing the pattern of among-site rate variation to change over time. Our investigations of genes from secondary and tertiary plastids derived from the rhodophyte plastid lineage clearly reveal "heterotachy" processes characterized as stationary covarion substitution patterns and changes in proportion of variable sites across sequences. Failure to accommodate covarion-like substitution patterns can have strong effects on the plastid tree topology. Importantly, multigene analyses performed with probabilistic methods using among-site rate and covarion models of evolution conflict with proposed single origin of the peridinin- and 19'-HNOF-containing plastids, suggesting that analysis of secondhand plastids can be hampered by convergence in the evolutionary signature of the plastid DNA sequences. Another type of sequence convergence was detected at protein level involving the psaA gene. Excluding the psaA sequence from a concatenated protein alignment grouped the peridinin plastid with haptophytes, congruent with all DNA trees. Altogether, taking account of complex processes involved in the evolution of dinoflagellate plastid sequences (both at the DNA and amino acid level), we demonstrate the difficulty of excluding independent, tertiary origin for both the peridinin and 19'-HNOF plastids involving engulfment of haptophyte-like algae. In addition, the refined topologies suggest the red algal order, Porphyridales, as the endosymbiont ancestor of the secondary plastids in cryptophytes, haptophytes, and heterokonts. PMID- 16699170 TI - A proton delivery pathway in the soluble fumarate reductase from Shewanella frigidimarina. AB - The mechanism for fumarate reduction by the soluble fumarate reductase from Shewanella frigidimarina involves hydride transfer from FAD and proton transfer from the active-site acid, Arg-402. It has been proposed that Arg-402 forms part of a proton transfer pathway that also involves Glu-378 and Arg-381 but, unusually, does not involve any bound water molecules. To gain further insight into the importance of this proton pathway we have perturbed it by substituting Arg-381 by lysine and methionine and Glu-378 by aspartate. Although all the mutant enzymes retain measurable activities, there are orders-of-magnitude decreases in their k(cat) values compared with the wild-type enzyme. Solvent kinetic isotope effects show that proton transfer is rate-limiting in the wild type and mutant enzymes. Proton inventories indicate that the proton pathway involves multiple exchangeable groups. Fast scan protein-film voltammetric studies on wild-type and R381K enzymes show that the proton transfer pathway delivers one proton per catalytic cycle and is not required for transporting the other proton, which transfers as a hydride from the reduced, protonated FAD. The crystal structures of E378D and R381M mutant enzymes have been determined to 1.7 and 2.1 A resolution, respectively. They allow an examination of the structural changes that disturb proton transport. Taken together, the results indicate that Arg-381, Glu-378, and Arg-402 form a proton pathway that is completely conserved throughout the fumarate reductase/succinate dehydrogenase family of enzymes. PMID- 16699171 TI - Distinct signaling functions for Shc isoforms in the heart. AB - Thrombin activates protease-activated receptor-1 (PAR-1) and engages signaling pathways that influence the growth and survival of cardiomyocytes as well as extracellular matrix remodeling by cardiac fibroblasts. This study examines the role of Shc proteins in PAR-1-dependent signaling pathways that influence ventricular remodeling. We show that thrombin increases p46Shc/p52Shc phosphorylation at Tyr(239)/Tyr(240) and Tyr(317) (and p66Shc-Ser(36) phosphorylation) via a pertussis toxin-insensitive epidermal growth factor receptor (EGFR) transactivation pathway in cardiac fibroblasts; p66Shc-Ser(36) phosphorylation is via a MEK-dependent mechanism. In contrast, cardiac fibroblasts express beta(2)-adrenergic receptors that activate ERK through a pertussis toxin-sensitive EGFR transactivation pathway that does not involve Shc isoforms or lead to p66Shc-Ser(36) phosphorylation. In cardiomyocytes, thrombin triggers MEK-dependent p66Shc-Ser(36) phosphorylation, but this is not via EGFR transactivation (or associated with Shc-Tyr(239)/Tyr(240) and/or Tyr(317) phosphorylation). Importantly, p66Shc protein expression is detected in neonatal, but not adult, cardiomyocytes; p66Shc expression is induced (via a mechanism that requires protein kinase C and MEK activity) by Pasteurella multocida toxin, a Galpha(q) agonist that promotes cardiomyocyte hypertrophy. These results identify novel regulation of individual Shc isoforms in receptor-dependent pathways leading to cardiac hypertrophy and the transition to heart failure. The observations that p66Shc expression is induced by a Galpha(q) agonist and that PAR-1 activation leads to p66Shc-Ser(36) phosphorylation identifies p66Shc as a novel candidate hypertrophy-induced mediator of cardiomyocyte apoptosis and heart failure. PMID- 16699172 TI - Structural analysis of protein kinase A mutants with Rho-kinase inhibitor specificity. AB - Controlling aberrant kinase-mediated cellular signaling is a major strategy in cancer therapy; successful protein kinase inhibitors such as Tarceva and Gleevec verify this approach. Specificity of inhibitors for the targeted kinase(s), however, is a crucial factor for therapeutic success. Based on homology modeling, we previously identified four amino acids in the active site of Rho-kinase that likely determine inhibitor specificities observed for Rho-kinase relative to protein kinase A (PKA) (in PKA numbering: T183A, L49I, V123M, and E127D), and a fifth (Q181K) that played a surprising role in PKA-PKB hybrid proteins. We have systematically mutated these residues in PKA to their counterparts in Rho-kinase, individually and in combination. Using four Rho-kinase-specific, one PKA specific, and one pan-kinase-specific inhibitor, we measured the inhibitor binding properties of the mutated proteins and identify the roles of individual residues as specificity determinants. Two combined mutant proteins, containing the combination of mutations T183A and L49I, closely mimic Rho-kinase. Kinetic results corroborate the hypothesis that side-chain identities form the major determinants of selectivity. An unexpected result of the analysis is the consistent contribution of the individual mutations by simple factors. Crystal structures of the surrogate kinase inhibitor complexes provide a detailed basis for an understanding of these selectivity determinant residues. The ability to obtain kinetic and structural data from these PKA mutants, combined with their Rho-kinase-like selectivity profiles, make them valuable for use as surrogate kinases for structure-based inhibitor design. PMID- 16699173 TI - Inhibitory effects of voluntary running wheel exercise on UVB-induced skin carcinogenesis in SKH-1 mice. AB - Earlier studies showed that oral administration of green tea or caffeine to SKH-1 mice inhibited ultraviolet B light (UVB)-induced skin carcinogenesis, decreased dermal fat thickness and increased locomotor activity. In the present study, the effects of voluntary running wheel exercise on thickness of dermal fat as well as on UVB-induced tumorigenesis in SKH-1 mice were studied in UVB-initiated high risk and UVB-induced complete carcinogenesis models. In the high-risk model, animals were exposed to UVB (30 mJ/cm(2)) 3 times/week for 16 weeks. For 14 weeks subsequent to UVB exposure, half of the animals had access to running wheels in their cages whereas the other half did not. In the complete carcinogenesis model, animals were exposed to UVB (30 mJ/cm(2)) 2 times/week for 33 weeks. From the beginning, half of the animals had access to running wheels whereas the other half did not. At the conclusion of each study, body weights were not different between groups, although animals with running wheels consumed significantly more food and water than animals without running wheels. In addition, animals with running wheels had decreases in parametrial fat pad weight and thickness of the dermal fat layer. In both UVB-initiated high-risk and complete carcinogenesis models, voluntary running wheel exercise delayed the appearance of tumors, decreased the number of tumors per mouse and decreased tumor volume per mouse. Histopathology studies revealed that running wheel exercise decreased the number of non-malignant tumors (primarily keratoacanthomas) by 34% and total tumors per mouse by 32% in both models, and running wheel exercise decreased the formation of squamous cell carcinomas in the UVB-induced complete carcinogenesis model by 27%. In addition, the size of keratoacanthomas and squamous cell carcinomas were decreased substantially in both models. The effects described here indicate that voluntary running wheel exercise inhibits UVB-induced skin tumorigenesis and may also inhibit tumor growth. PMID- 16699174 TI - The necessity of a human epigenome project. AB - Epigenetics is one of the hottest topics in cancer research. We know that human tumors undergo a major disruption of their DNA methylation and histone modification patterns. The aberrant epigenetic landscape of the cancer cell is characterized by a massive genomic hypomethylation, CpG island promoter hypermethylation of tumor suppressor genes, an altered histone code for critical genes and a global loss of monoacetylated and trimethylated histone H4. But what we know is just a minimal percentage of the epigenetic 'earthquake' present in the transformed cell. We need to make an ambitious step to understand the DNA methylation and histone changes underlying tumorigenesis. The launching of an International Human Epigenome Project should be the response to this necessity. PMID- 16699175 TI - Negative energy balance induced by voluntary wheel running inhibits polyp development in APCMin mice. AB - Treadmill running of approximately 0.9 km/day has had inconsistent effects on spontaneous intestinal polyp development in C57BL/6J-Apc(Min)/J (Min) mice; the amount of energy expenditure and/or a lack of hormonal changes could account for this variability. The purpose of this study was to examine the effects of a negative energy balance induced by voluntary wheel running on polyps, insulin like growth factor-1 (IGF-1) and corticosterone in Min mice. Seven-week-old male Min mice were randomly assigned to control (CON, n = 23) or wheel running (EX, n = 24) conditions for a 10-week study period. All mice had water and AIN-76A diet ad libitum for the first approximately 3 weeks on study, after which the EX group was pair-fed to the CON group to maintain a negative energy balance due to the exercise. EX mice voluntarily ran 3.8 km/day (2.7-6.0 km/day) (median, interquartile range) and weighed less than CON mice throughout the study. More CON mice died before the end of the study versus EX mice (26 versus 0%, P < 0.01). CON mice had significantly more polyps versus EX mice (21.6 +/- 1.5 versus 16.9 +/- 2.0, P < 0.01; mean +/- SE), and daily running distance in EX was inversely correlated with total polyp number (r = -0.70, P < 0.01). Urinary corticosterone output (P < 0.01) and serum IGF-1 were significantly higher in EX than CON (P < 0.001); however, total polyp number was unrelated to corticosterone (r = 0.05, P = 0.84) and IGF-1 (r = -0.01, P = 0.93). In this study, a negative energy balance produced by wheel running exercise and restricted feeding decreased polyp burden in male Min mice and appeared to have a dose-response effect on polyp number. Although EX affected IGF-1 and corticosterone, neither marker was related to total polyp number. PMID- 16699176 TI - In diatoms, the transthylakoid proton gradient regulates the photoprotective non photochemical fluorescence quenching beyond its control on the xanthophyll cycle. AB - In diatoms, the non-photochemical fluorescence quenching (NPQ) regulates photosynthesis during light fluctuations. NPQ is associated with an enzymatic xanthophyll cycle (XC) which is controlled by the light-driven transthylakoid proton gradient (delta pH). In this report, special illumination conditions and chemicals were used to perturb the kinetics of the delta pH build-up, of the XC and of NPQ. We found that the delta pH-related acidification of the lumen is also needed for NPQ to develop by switching the xanthophylls to an 'activated' state, probably via the protonation of light-harvesting antenna proteins. It confirms the NPQ model previously proposed for diatoms. PMID- 16699177 TI - Genetic interaction between the AS1-AS2 and RDR6-SGS3-AGO7 pathways for leaf morphogenesis. AB - In higher plants, class I KNOTTED1-like homeobox (KNOX) gene suppression and leaf polarity establishment are two processes crucial for leaf morphogenesis. The Arabidopsis genes, ASYMMETRIC LEAVES1 and 2 (AS1 and AS2), are required for repressing the class I KNOX genes and promoting leaf adaxial cell fates. In addition, the RNA-DEPENDENT RNA POLYMERASE6 (RDR6) gene acts synergistically with AS1 and AS2 to specify the adaxial polarity and repress the KNOX genes in leaves. It is known that RDR6 is one of the key components in plant post-transcriptional gene silencing (PTGS), and is likely to function with other silencing components in a genetic pathway in regulating leaf patterning. Here we report phenotypic analyses of double mutants combining as1 or as2 with other mutations relating to different RNA silencing pathways. We show that plants carrying rdr6, suppressor of gene silencing3 (sgs3) or zippy (zip, also called ago7) in combination with as1 or as2 demonstrate severe morphological defects, and the double mutant plants are generally similar to one another. Detailed phenotypic and molecular analyses reveal that leaves of rdr6 as2(1), sgs3 as2(1) and zip as2(1) all show an abnormal adaxial identity, and contain high levels of microRNA165/166 and FILAMENTOUS FLOWER (FIL) transcripts. These results suggest that RDR6, SGS3 and AGO7 act in the same pathway, which genetically interacts with the AS1-AS2 pathway for leaf development. The RDR6-SGS3-AGO7 pathway was previously identified as regulating the plant vegetative phase change. Our results reveal a new function of the pathway, which is also required for normal leaf morphogenesis. PMID- 16699178 TI - n-Butanol induces depolymerization of microtubules in vivo and in vitro. AB - The effects of butanol on microtubules (MTs) were examined by immunofluorescence microscopy. Fragmentation of cortical MTs was induced by n-butanol, but not by s- and t-butanols, in cultured tobacco BY-2 cells. Taxol prevented n-butanol-induced MT fragmentation. Fragmented cortical MTs were still attached to the inner face of the plasma membrane when n-butanol-treated protoplasts were ruptured on the slide glass. Moreover, MTs were depolymerized in the presence of n-butanol in vitro. Therefore, n-butanol is not only an activator of phospholipase D but also an effective MT-depolymerizing agent. PMID- 16699179 TI - Identification of a Sed5-like SNARE gene LjSYP32-1 that contributes to nodule tissue formation of Lotus japonicus. AB - We identified a Sed5-like clone LjSYP32-1 which contributes to nodule tissue formation and plant growth in Lotus japonicus. In the L. japonicus expressed sequence tag (EST) clone databases of Kazusa DNA Research Institute, another syntaxin-related clone (LjSYP32-2) was also detected, and the nucleotide and amino acid sequences of these two clone are very similar to each other. Real-time PCR and promoter analysis indicated that expression of LjSYP32-1 was dominant compared with LjSYP32-2 in the various plant organs. Promoter analysis and in situ hybridization revealed that LjSYP32-1 was expressed significantly in the inner cortex cell layer surrounding the infected zone of young nodules and in the meristem area of developing lateral root. To explore the function and physiological role of LjSYP32-1 in nodules and other plant organs, stable transformation lines of L. japonicus expressing either sense or antisense LjSYP32 1 were prepared. The antisense plants showed a significantly retarded plant growth phenotype, suggesting a role for LjSYP32-1 in supporting plant growth. In the same transgenic lines, the plants were capable of forming nodules, but the acetylene reduction activity was reduced by around 50% per plant. The nodules were much smaller and some nodules were fused to each other by sharing the inner cortex. The rate of occurrence of such irregular nodules was twice that observed in wild-type plants. The data suggest that LjSYP32-1 contributes to the support of plant growth and normal nodule tissue differentiation. PMID- 16699180 TI - The adenomatous polyposis coli tumor suppressor gene regulates expression of cyclooxygenase-2 by a mechanism that involves retinoic acid. AB - Mutations in the adenomatous polyposis coli (APC) gene result in uncontrolled proliferation of intestinal epithelial cells and are associated with the earliest stages of colorectal carcinogenesis. Cyclooxygenase-2 (COX-2) is elevated in human colorectal cancers and plays an important role in colorectal tumorigenesis; however, the mechanisms by which APC mutations result in increased COX-2 expression remain unclear. We utilized APC mutant zebrafish and human cancer cells to investigate how APC modulates COX-2 expression. We report that COX-2 is up-regulated in APC mutant zebrafish because of a deficiency in retinoic acid biosynthesis. Treatment of both APC mutant zebrafish and human carcinoma cell lines with retinoic acid significantly reduces COX-2 expression. Retinoic acid regulates COX-2 levels by a mechanism that involves participation of the transcription factor C/EBP-beta. APC mutant zebrafish express higher levels of C/EBP-beta than wild-type animals, and retinoic acid supplementation reduces C/EBP-beta expression to basal levels. Both morpholino knockdown of C/EBP-beta in APC mutant zebrafish and silencing of C/EBP-beta using small interfering RNA in HT29 colon cancer cells robustly decrease COX-2 expression. Our findings support a sequence of events in which mutations in APC result in impaired retinoic acid biosynthesis, elevated levels of C/EBP-beta, up-regulation of COX-2, increased prostaglandin E(2) accumulation, and activation of Wnt target genes. PMID- 16699182 TI - Crystallization to obtain protein-ligand complexes for structure-aided drug design. AB - The use of X-ray crystallography to derive three-dimensional structures for structure-aided drug design (SADD) is a common activity in drug discovery today. In this process, the structures of inhibitors or other ligands of interest complexed with their macromolecular target are solved and the structural information is used iteratively to design new molecules. The ability to form cocrystal complexes between a target protein and a ligand is essential to this process and therefore is of considerable interest to anyone practicing in this field. In the course of obtaining the necessary ligand-protein crystals, even with crystallization conditions well established for a protein of interest, obtaining co-structures with inhibitors either through cocrystallization or soaking is too often not successful. There are numerous potential reasons for this lack of success and this article outlines a number of possible factors that may be involved and discusses considerations that should be taken into account when designing successful experiments to obtain iterative costructures. PMID- 16699181 TI - Identification and solution structures of a single domain biotin/lipoyl attachment protein from Bacillus subtilis. AB - Protein biotinylation and lipoylation are post-translational modifications, in which biotin or lipoic acid is covalently attached to specific proteins containing biotin/lipoyl attachment domains. All the currently reported natural proteins containing biotin/lipoyl attachment domains are multidomain proteins and can only be modified by either biotin or lipoic acid in vivo. We have identified a single domain protein with 73 amino acid residues from Bacillus subtilis strain 168, and it can be both biotinylated and lipoylated in Escherichia coli. The protein is therefore named as biotin/lipoyl attachment protein (BLAP). This is the first report that a natural single domain protein exists as both a biotin and lipoic acid receptor. The solution structure of apo-BLAP showed that it adopts a typical fold of biotin/lipoyl attachment domain. The structure of biotinylated BLAP revealed that the biotin moiety is covalently attached to the side chain of Lys(35), and the bicyclic ring of biotin is folded back and immobilized on the protein surface. The biotin moiety immobilization is mainly due to an interaction between the biotin ureido ring and the indole ring of Trp(12). NMR study also indicated that the lipoyl group of the lipoylated BLAP is also immobilized on the protein surface in a similar fashion as the biotin moiety in the biotinylated protein. PMID- 16699183 TI - The high-resolution structure of (+)-epi-biotin bound to streptavidin. AB - (+)-Epi-biotin differs from (+)-biotin in the configuration of the chiral center at atom C2. This could lead to a difference in the mode of binding of (+)-epi biotin to streptavidin, a natural protein receptor for (+)-biotin. Diffraction data were collected to a maximum of 0.85 Angstrom resolution for structural analysis of the complex of streptavidin with a sample of (+)-epi-biotin and refinement was carried out at both 1.0 and 0.85 Angstrom resolution. The structure determination shows a superposition of two ligands in the binding site, (+)-biotin and (+)-epi-biotin. The molecules overlap in the model for the complex except for the position of S1 in the tetrahydrothiophene ring. Differences in the conformation of the ring permits binding of each molecule to streptavidin with little observable difference in the protein structures at this high resolution. PMID- 16699184 TI - Structure of human DSP18, a member of the dual-specificity protein tyrosine phosphatase family. AB - The human dual-specificity protein phosphatase 18 (DSP18) gene and its protein product have recently been characterized. Like most DSPs, DSP18 displays dephosphorylating activity towards both phosphotyrosine and phosphothreonine residues. However, DSP18 is distinct from other known DSPs in terms of the existence of approximately 30 residues at the C-terminus of the catalytic domain and an unusual optimum activity profile at 328 K. The crystal structure of human DSP18 has been determined at 2.0 A resolution. The catalytic domain of DSP18 adopts a fold similar to that known for other DSP structures. Although good alignments are found with other DSPs, substantial differences are also found in the regions surrounding the active site, suggesting that DSP18 constitutes a unique structure with a distinct substrate specificity. Furthermore, the residues at the C-terminus fold into two antiparallel beta-strands and participate in extensive interactions with the catalytic domain, explaining the thermostability of DSP18. PMID- 16699185 TI - Structural elucidation of the PDI-related chaperone Wind with the help of mutants. AB - The structures of the PDI-related protein Wind (with a C-terminal His(6) tag) and the mutants Y53S, Y53F and Y55K have been determined and compared with the wild type structure with the His(6) tag at the N-terminus. All five structures show the same mode of dimerization, showing that this was not an artefact introduced by the nearby N-terminal His(6) tag and suggesting that this dimer may also be the biologically active form. Although the mutants Y53S and Y55K completely abrogate transport of the protein Pipe (which appears to be the primary function of Wind in the cell), only subtle differences can be seen in the putative Pipe binding region. The Pipe binding in the active forms appears to involve hydrophobic interactions between aromatic systems, whereas the inactive mutants may be able to bind more strongly with the help of hydrogen bonds, which could disturb the delicate equilibrium required for effective Pipe transport. PMID- 16699186 TI - Structure of recombinant Ves v 2 at 2.0 Angstrom resolution: structural analysis of an allergenic hyaluronidase from wasp venom. AB - Wasp venom from Vespula vulgaris contains three major allergens: Ves v 1, Ves v 2 and Ves v 5. Here, the cloning, expression, biochemical characterization and crystal structure determination of the hyaluronidase Ves v 2 from family 56 of the glycoside hydrolases are reported. The allergen was expressed in Escherichia coli as an insoluble protein and refolded and purified to obtain full enzymatic activity. Three N-glycosylation sites at Asn79, Asn99 and Asn127 were identified in Ves v 2 from a natural source by enzymatic digestions combined with MALDI-TOF mass spectrometry. The crystal structure of recombinant Ves v 2 was determined at 2.0 A resolution and reveals a central (beta/alpha)(7) core that is further stabilized by two disulfide bonds (Cys19-Cys308 and Cys185-Cys197). Based on sequence alignments and structural comparison with the honeybee allergen Api m 2, it is proposed that a conserved cavity near the active site is involved in binding of the substrate. Surface epitopes and putative glycosylation sites have been compared with those of two other major group 2 allergens from Apis mellifera (honeybee) and Dolichovespula maculata (white-faced hornet). The analysis suggests that the harboured allergic IgE-mediated cross-reactivity between Ves v 2 and the allergen from D. maculata is much higher than that between Ves v 2 and the allergen from A. mellifera. PMID- 16699187 TI - A survey of protein-protein complex crystallizations. AB - A survey of crystallization conditions was carried out for 650 published protein protein complexes in the Protein Data Bank (PDB) of the Research Collaboratory for Structural Bioinformatics (RCSB). This resulted in the establishment of a Protein Complex Crystallization Database (PCCD) and a set of configuration-space boundaries for protein-complex crystallizations. Overall, polyethylene glycol (PEG) based conditions accounted for 70-80% of all crystallizations, with PEG 3000-4000, 5000-6000 and 8000 being the most frequently used. The median values of PEG concentrations were between 10 and 20% and were inversely correlated with their molecular weights. Ammonium sulfate remained the most favorable salt precipitant, with a median concentration of 1.6 M. The crystallization pH for the vast majority of protein complexes was between 5.0 and 8.0. Overall, the boundaries for the crystallization configuration space of protein complexes appear to be more restricted than those of soluble proteins. This may reflect the limited stability and solubility of protein-protein complexes. Based on statistical analysis of the database, a sparse-matrix and a systematic buffer and pH screen were formulated to best represent the crystallization of protein complexes. PMID- 16699188 TI - Mapping the active site of Escherichia coli malonyl-CoA-acyl carrier protein transacylase (FabD) by protein crystallography. AB - Malonyl-CoA-acyl carrier protein transacylase (FabD; EC 2.3.1.39) is a key enzyme in the fatty-acid biosynthesis pathway of bacteria, catalyzing the transfer of a malonyl moiety from malonyl-CoA to holo acyl carrier protein (ACP), generating malonyl-ACP and free CoASH. Malonyl-ACP, which is the product of this reaction, is the key building block for de novo fatty-acid biosynthesis. Various binary complex structures of the Escherichia coli enzyme are presented, including that of the natural substrate malonyl-CoA, indicating the functional role of the highly conserved amino acids Gln11, Ser92, Arg117 and His201 and the stabilizing function of the preformed oxyanion hole during the enzymatic reaction. Based on the presented structural data, a possible new catalytic enzyme mechanism is discussed. The data obtained could be used in aiding the process of rational inhibitor design. PMID- 16699189 TI - Novel crystal form of the ColE1 Rom protein. AB - The RNA I modulator protein (Rom) acts as a co-regulator of ColE1 plasmid copy number by binding to RNA kissing hairpins and stabilizing their interaction. The structure of Rom has been determined in a new crystal form from X-ray diffraction data to 2.5 A resolution. In this structure, a dimer of the 57-amino-acid protein is found in the asymmetric unit. Each subunit consists almost entirely of two antiparallel alpha-helices joined by a short hairpin bend. The dimer contains a non-crystallographic twofold axis and forms a highly regular four-alpha-helical bundle. The structural packing in this novel crystal form is different from previously known Rom structures. The asymmetric unit contains one dimer, giving a crystal volume per protein weight (V(M)) of 1.83 A(3) Da(-1) and a low solvent content of 30%. Strong packing interactions and low solvation are characteristic of the structure. The Rom protein was cocrystallized with the Tar-Tar* kissing hairpin RNA. Although the electron-density maps do not show bound RNA, altered conformations in the side chains of Rom that are known to be involved in RNA binding have been identified. These results provide additional information about Rom protein conformational flexibility and suggest that the presence of a highly charged polymer such as RNA can promote tight packing of an RNA-binding protein, even when the RNA itself is not observed in the crystal. PMID- 16699190 TI - Invariance and variability in bacterial PanK: a study based on the crystal structure of Mycobacterium tuberculosis PanK. AB - Pantothenate kinase (PanK) is a ubiquitous and essential enzyme that catalyzes the first step of the universal coenzyme A biosynthetic pathway. In this step, pantothenate (vitamin B(5)) is converted to 4'-phosphopantothenate, which subsequently forms coenzyme A in four enzymatic steps. The complex of this enzyme from Mycobacterium tuberculosis (MtPanK) with a derivative of the feedback inhibitor coenzyme A has been crystallized in two forms and its structure solved. The structure was refined in both forms using room-temperature and low temperature X-ray data. In both forms, the MtPanK subunit has a mononucleotide binding fold with a seven-stranded central beta-sheet and helices on either side. However, there is a small though significant difference in subunit association between the two forms. The structure is also grossly similar to the enzyme from Escherichia coli. The active-site pocket and the dimeric interface are on two opposite sides of the PanK subunit. The enzymes from M. tuberculosis and E. coli exhibit several differences, particularly at the dimeric interface. On the other hand, the coenzyme A-binding region is almost entirely conserved. A delineation of the invariant and variable features of the PanK structure further indicates that the dimeric interface is very variable, while the coenzyme A-binding site is substantially invariant. A sequence alignment involving various bacterial PanKs is in agreement with this conclusion. The strong correlation between structural plasticity, evolutionary conservation and variability and function exhibited by the molecule could be important in the design of species-specific inhibitors of the enzyme. PMID- 16699191 TI - Interaction energies between glycopeptide antibiotics and substrates in complexes determined by X-ray crystallography: application of a theoretical databank of aspherical atoms and a symmetry-adapted perturbation theory-based set of interatomic potentials. AB - Intermolecular interaction energies between fragments of glycopeptide antibiotics and small peptide ligands are evaluated using geometries determined by X-ray crystallography and recently developed methods suitable for application to very large molecular complexes. The calculation of the electrostatic contributions is based on charge densities constructed with a databank of transferable aspherical atoms described by nucleus-centered spherical harmonic density functions, and uses the accurate and fast EPMM method. Dispersion, induction and exchange repulsion contributions are evaluated with atom-atom potentials fitted to intermolecular energies from SAPT (symmetry-adapted perturbation theory) calculations on a large number of molecules. For a number of the complexes, first principle calculations using density functional theory have been performed for comparison. Results of the new methods agree within reasonable bounds with those from DFT calculations, while being obtained at a fraction (less than 1%) of the computer time. A strong dependence on the geometry of the interaction is found, even when the number of hydrogen bonds between the substrate and antibiotic fragment is the same. While high-resolution X-ray data are required to obtain interaction energies at a quantitative level, the techniques developed have potential for joint X-ray/energy refinement of macromolecular structures. PMID- 16699192 TI - MUMBO: a protein-design approach to crystallographic model building and refinement. AB - In recent years, significant progress has been achieved in automation of the crystal structure-determination process. However, the final part of this process, namely the refinement of the atomic model, is still tedious for biological macromolecules because, especially at lower resolution, it requires extensive manual intervention. Here, it is shown that computer algorithms widely used in protein-design approaches can substantially simplify this process, helping to identify the correct orientation of the side chains during refinement. This approach was implemented into the computer program MUMBO. As in many protein design programs, side-chain rotamer diversity is generated using rotamer libraries. The selection of the best combination of side-chain orientations is based on either the dead-end elimination (DEE) theorem or a Metropolis Monte Carlo approach and on a detailed atomic scoring function that describes the molecular interactions between the rotamers. We show that this scoring function can be easily extended and complemented through the introduction of an X-ray pseudo-energy calculated from the electron density present at the position of the rotamer. This extension is fully compatible with present protein-design algorithms and it is shown for a number of test cases that using this approach model refinement is simplified and convergence occurs faster. PMID- 16699193 TI - The first crystal structure of an RNA racemate. AB - The racemate of the RNA duplex r(CUGGGCGG).r(CCGCCUGG) from Thermus flavus 5S rRNA has been crystallized and examined by X-ray crystallography. The space group is P1(_) with approximate unit-cell parameters a = 26.5, b = 38.0, c = 45.4 Angstrom, alpha = 113.1, beta = 100.5, gamma = 93.3 degrees. The structure was solved by molecular replacement. There are four RNA duplexes in the unit cell. The crystal lattice consists of columns of RNA duplexes. The duplexes are stacked end-to-end and are stabilized by intermolecular base-stacking interactions. Within each column the L-duplexes and D-duplexes are stacked alternately. Every other duplex in each stack has two alternative conformations, approximately equally occupied, corresponding to molecules oriented in opposite directions. Neighbouring columns are related by the crystallographic centre of symmetry. The unit cell also contains approximately 250 ordered water molecules and six ordered calcium ions. A glycerol molecule is visible in the minor groove interacting with a guanosine residue. PMID- 16699194 TI - Multiple crystal forms of the cell-wall invertase inhibitor from tobacco support high conformational rigidity over a broad pH range. AB - Plant acid invertases catalyse the breakdown of sucrose. Their activity is tightly regulated through interaction with specific protein inhibitors. The complex between the cell-wall invertase inhibitor Nt-CIF and its target enzyme is stable only at acidic pH, as found in the plant cell wall. Since the pH in this compartment can be modulated between pH 4 and 6 in planta, the rapid dissociation of the inhibitor-enzyme complex at neutral pH may represent a regulatory event. Here, it is analyzed whether the inhibitory component undergoes structural rearrangements upon changes in the pH environment. Six crystal forms grown at pH 4.6-9.5 and diffracting up to 1.63 Angstrom indicate only small structural changes in CIF. This suggests that complex dissociation at neutral pH is mediated either by rearrangements in the enzyme or by a complex pattern of surface charges in the inhibitor-enzyme binding interface. PMID- 16699195 TI - High-resolution structure of human cytoglobin: identification of extra N- and C termini and a new dimerization mode. AB - Cytoglobin (Cgb) is a recently discovered member of the vertebrate haem containing globin family. The structure of a new crystal form of wild-type human Cgb (space group C2) was determined at a resolution of 1.68 Angstrom. The results show the presence of an additional helix in the N-terminal residues (4-20) prior to the A helix and an ordered loop structure in the C-terminal region (168-188), while these extended peptides were invisible owing to disorder in the previously reported structures using a P3(2)21 crystal at a resolution of 2.4 Angstrom. A detailed comparison of the two crystal structures shows differences in the conformation of the residues (i.e. Arg84) in the haem environment owing to a different dimeric arrangement. PMID- 16699196 TI - Small revisions to predicted distances around metal sites in proteins. AB - A new analysis has been made of distances around metal sites in protein structures in the Protein Data Bank determined with resolution < or =1.25 A and equivalent distances have been extracted from the Cambridge Structural Database. They are for the metals Na, Mg, K, Ca, Mn, Fe, Co, Cu, Zn and the donor atoms O of water, O of Asp and Glu, O of the main-chain carbonyl group, N of His and S of Cys. Some revisions are recommended to the tables of 'target distances' previously given. As well as small changes in many distances and a large improvement for Mg-O(carboxylate), the table includes an indication of how reliable each prediction may be. Special attention was given to carboxylate interactions. When the carboxylate group is monodentate, the M-O(carboxylate) distance is well defined, but for bidentate carboxylate groups a wide range of distances is allowable; when the metal is Co, Cu or Zn the M-O(1) and M-O(2) distances are clearly inversely correlated; for the more purely electrostatic interactions involving Na, K and Ca there is a wider scatter of distances and little correlation. PMID- 16699198 TI - Measurements of emittance and absolute spectral flux of the PETRA undulator at DESY Hamburg. AB - The first synchrotron radiation beamline using a 4 m-long undulator at the 12 GeV storage ring PETRA delivers hard X-ray photons usable up to 300 keV. The photon intensity is measured on an absolute scale in the energy range between 16 and 60 keV and compared with calculated intensities. The experimental set-up described is also used to measure the horizontal and vertical emittance of the source. PMID- 16699197 TI - Structure determination and analysis of acyl-CoA oxidase (ACX1) from tomato. AB - The flavoenzyme acyl-CoA oxidase (ACX) catalyzes the first committed step in beta oxidation and is required for the biosynthesis of jasmonic acid, a signaling molecule involved in plant defense. Recently, a mutant in tomato was identified that is deficient in jasmonic acid production and compromised in its wound response. This results from a single point mutation in acx1, which causes the conserved residue Thr138 to be substituted by isoleucine. To understand the structural basis for this mutation, the crystal structure of LeACX1 was determined to 2.74 Angstrom resolution by molecular replacement. Unexpectedly, an unusual packing arrangement was observed in which three monomers of LeACX1 are present in the asymmetric unit. Although the tertiary structure of LeACX1 is essentially similar to the previously determined structures of ACX enzymes, the packing within the unit cells is distinctly different. PMID- 16699199 TI - Development of a tunable UV laser system synchronizing precisely with synchrotron radiation pulses from UVSOR. AB - A mode-locked Ti:sapphire laser is made to oscillate at the frequency of the UVSOR storage ring, 90.115 MHz, in a multi-bunch operation mode. The third harmonic of the laser is available in the wavelength range 243-280 nm. Synchrotron radiation from an undulator is monochromated by a grazing-incidence monochromator and introduced coaxially with the laser. The temporal profile of the photon pulses is monitored in situ by a luminescing substance/photomultiplier combination. The delay timing between the laser and synchrotron radiation can be changed from 0 to 11 ns by adjusting an electronic module that provides phase locked loop stabilization of the laser pulse. The reliability and feasibility of this laser-synchrotron radiation combination technique are demonstrated by applying pump-probe experiments to two physical systems. The first system is photodissociation of iodomethane (CHA) with a laser photon, followed by photoionization of I and CH3 fragments with synchrotron radiation. The second, two-photon ionization of He atoms, is studied as the prototype of a time-resolved experiment. The He+ signal counts as a function of the laser-synchrotron radiation delay are found to be enhanced in a narrow time window, which can be interpreted in terms of a short lifetime of the resonant state, He*(1s2p 1P), produced by primary synchrotron radiation excitation. PMID- 16699200 TI - Time-resolved protein crystallography with large-angle oscillations: an application of a protein data-collection system using the Weissenberg technique and a large-format imaging plate. AB - A diffraction-intensity data-collection system with synchrotron radiation X-rays utilizing the screenless Weissenberg technique and incorporating a large-format imaging plate is one of the most suitable apparatus for time-resolved protein crystallography with larger angle oscillations than hitherto described. The time resolution and data quality of the system have been tested using a tetragonal lysozyme crystal as a test sample in a flow-cell experiment at the bending-magnet beamline 18B at the Photon Factory, and a time resolution of 15 min is confirmed. PMID- 16699201 TI - Protein crystallography in the soft X-ray region: crystal lifetime and diffraction efficiency. AB - Multiwavelength anomalous diffraction on the sulfur (5.01 A) and phosphorus (5.77 A) absorption K-edges seems to offer a solution to one of the most appealing problems of protein crystallography - the determination of the phases of structure amplitudes. However, a strong increase in absorption of X-rays in this region may impede the development of this method of structure solution. Analytical expressions for diffraction efficiency and normalized diffraction efficiency have been derived in the present paper. It is shown that the crystal lifetime in the soft X-ray region will be significantly shorter than the lifetime of a macromolecular crystal exposed to 1-1.5 A X-rays during diffraction data collection. An optimum crystal size has been estimated. PMID- 16699202 TI - Application of an imaging plate to the large-volume press MAX80 at the Photon Factory. AB - An imaging plate was applied to the large-volume press MAX80 for quantitative in situ X-ray diffraction. A combination of the two-dimensional detector with the uniform pressure and temperature environments of MAX80 and the wide energy range of synchrotron radiation resulted in high-quality diffraction data. A modified cell assembly with a disc-type heater was used to minimize extrinsic diffraction peaks from the surrounding materials. To demonstrate the high-quality data obtained from the imaging-plate system, the crystal structure of the high pressure and high-temperature polymorph of the highly absorbing element bismuth (Bi IV) was determined. PMID- 16699203 TI - X-ray fluorescence used to characterize the salt content of proteins. AB - The quantitative measurement of the salt content in solid protein samples was performed using X-ray fluorescence. Linear calibration curves were obtained for chloride, calcium and sulfur using sulfur and chloride as internal standards in the range 1-10 protein molar equivalents. The detection limit was approximately 0.02 molar equivalents for chloride and less than 0.01 molar equivalents for calcium. X-ray fluorescence thus provides a non-destructive sensitive method of testing the efficiency of different purification methods. Commercial hen egg white lysozyme samples contain from 15 to 46 molar equivalents of chloride, whereas the calcium content remains less than 0.2 equivalents. Deionization on ion-exchange resins is a very efficient tool for removing ionic species since deionized lysozyme samples contain less than 0.34 molar equivalents of chloride. Extensive dialysis against water only partially removes chloride ions, the residual chloride content corresponding to the number of counter-ions necessary to ensure the electroneutrality of lysozyme when dissolved in water. PMID- 16699204 TI - X-FINE--a refinement program for FEFF input parameters. AB - X-FINE is an interactive and user-friendly tool to optimize input parameters (S20, sigma2, DeltaE) of the FEFF program, a package for EXAFS analysis. PMID- 16699206 TI - A method to provide magnet-free long straight sections in low-emittance synchrotron radiation sources. AB - In a low-emittance synchrotron radiation source it is difficult to provide magnet free long straight sections (MLSSs) a few tens of metres long because introducing MLSSs to the radiation source causes symmetry breaking and markedly reduces dynamic stability. To overcome this difficulty and realize MLSSs in such a ring, a method of building MLSSs is proposed starting from the stable lattice with high symmetry and proceeding via a transient lattice. PMID- 16699207 TI - Operation of an X-ray undulator in the Tristan main ring as a prototype of future super light sources. AB - A 5.4 m-long 239-pole undulator comprising three rigid and precise standardized unit undulators was completed and installed in the Tristan main ring, which was operated as a light source at a beam energy from 8 to 10 GeV. This undulator was successfully tested, characterized and used as a brilliant hard X-ray source for several synchrotron radiation experiments during the last 3.5 months of 1995. PMID- 16699208 TI - Hard X-ray microbeam experiment at the Tristan main ring test beamline of the KEK. AB - A hard X-ray microbeam with zone-plate optics has been tested at the MR-BW-TL beamline on the Tristan main ring of the KEK, and preliminary experiments on scanning microscopy have also been performed. A sputtered-sliced Fresnel zone plate with an Au core and Ag/C multilayer is used as an X-ray focusing device. The outermost zone width of the zone plate is 0.25 microm. A focused spot size of approximately 0.5 mum has been achieved at an X-ray energy of 8.54 keV. In a scanning X-ray microscopy experiment, test patterns with submicrometer fine structure have been clearly resolved. PMID- 16699209 TI - Casting technique for the fabrication of pinholes for X-ray radiation. AB - Pinholes with diameters down to 4 mum have been made in gold plates of a few hundred mum thickness. The fabrication method used is based on a casting technique. The pinholes are well suited for the collimation of (hard) X-rays from a synchrotron radiation source. PMID- 16699210 TI - Calibration and application of an X-ray image intensifier/ charge-coupled device detector for monochromatic macromolecular crystallography. AB - Charge-coupled device (CCD)-based X-ray detectors allow data to be collected much more quickly (approximately 10 times) than with current on-line imaging-plate systems. At the ESRF, X-ray image intensifier/CCD detector systems have been developed. These have great potential as fast read-out detectors for macromolecular and other forms of crystallography. They are relatively large sensitive X-ray detectors but have two inherent weaknesses: convex detection surfaces leading to spatial distortion and non-uniformity of intensity response, and susceptibility to small changes in magnetic fields. A large improvement has been made to the accuracy obtained by non-uniformity of response calibration and correction, using fluorescence from doped lithium borate glasses. Monochromatic macromolecular crystallography demonstration experiments with external user groups have shown that high-quality results may be obtained under real experimental conditions. PMID- 16699211 TI - Crystal structure study of (E)-3-(4-Nitrobenzyloxyimino)hexahydroazepin-2-one using synchrotron radiation. AB - NOHA, 3-(4-nitrobenzyloxyimino)hexahydroazepin-2-one, is a potent anticonvulsive agent. It occurs in two stereoisomers, the Z and E forms. (E)-NOHA is about 1.5 times more active than (Z)-NOHA. While accurate structural data are available for (Z)-NOHA, those for the E form have remained, until recently, highly speculative due to the lack of single crystals of suitable size for X-ray analysis and to the instability of the E isomer under radiation exposure. For structure-activity correlation purposes, efforts have been made to solve the crystal structure of the E isomer. Data collection from tiny needle crystals has been performed using synchrotron radiation. An initial molecular-packing model of this compound was obtained by energy-based and X-ray data modelling and successfully refined by molecular-dynamics methods: space group P2(1)/c, a = 6.225 (5), b = 17.885 (5), c = 12.157 (5) A, beta = 92.35 (5) degrees , R-factor = 3.5% for 343 reflections. In this contribution the crystal and molecular structure of (E)-NOHA is reported and the role of the hydrogen bond acting as the driving force in the intermolecular assembly in the (E)-NOHA crystal is highlighted. PMID- 16699212 TI - Time-resolved X-ray powder diffraction using a large-area CCD-based detector and Rietveld refinement: solid-state polymerization of S2N2 to (SN)x. AB - A kinetic study of the solid-state polymerization of disulfur dinitride (S2N2) to polysulfur nitride [(SN)x] has been performed, combining monochromatic high energy (lambda = 0.3263 A) synchrotron radiation X-ray powder diffraction, a large-area (o = 220 mm) CCD-based X-ray image-intensifier detector and Rietveld refinement. Recently developed techniques for detector calibration and reduction of two-dimensional images to one-dimensional diffraction patterns have been employed for data processing/analysis. Good fits were obtained after Rietveld refinement [Rp = 8.4%, wRp = 9.4%, sin(theta(max))/lambda = 0.585 A(-1)] of diffraction patterns of S2N2 from images with 2 s exposure time. The solid-state polymerization of S2N2 to (SN)x, was followed at a maximum rate of two diffraction images per minute. Scale factors and cell parameters for S2N2 and beta-(SN), as functions of time were readily obtained after Rietveld refinement of the diffraction patterns obtained from each individual image throughout the polymerization. The polymerization was preceded by a lattice distortion of S2N2, and at 50% conversion the a axis had decreased by about 1% and the c axis had increased about 1%. The polymerization yielded not only the expected polymer beta (SN)x, but also a small amount of a compound that could be another phase of (SN)x. PMID- 16699213 TI - Laue orientation imaging. AB - Advantage was taken of the highly focused X-ray beam (10-30 microm) and the broad white spectrum of synchrotron X-rays at the ESRF for automatic recording of Laue patterns from polycrystals and extraction of orientation information. The procedure used is similar to that applied for electron-backscattering patterns in the scanning electron microscope and provides data for local orientation mapping used in texture analysis. Laue patterns are obtained from a thin slice of material in transmission and recorded with a CCD detector. The Laue geometry is converted into a gnomonic projection in which co-zonal reflections lie on straight lines. On applying the Hough transform these lines are merged into a single point, which is recognized by the computer and assigned zone indices [uvw] by comparison with a table of interzonal angles. From the angular positions of several [uvw] the crystal orientation is calculated. The method is illustrated for the orthorhombic magnesium silicate olivine. PMID- 16699214 TI - High-energy magnetic compton scattering from iron. AB - The magnetic Compton profile of Fe [111] was measured using circularly polarized synchrotron radiation at incident energies of 84.4, 167.2 and 256.0 keV on the high-energy beamline at the European Synchrotron Radiation Facility. It was found that the momentum resolution of these experiments, which use semiconductor detectors, improves by almost a factor of two over what was previously possible by this technique at photon energies of approximately (1/10)mc(2). It was also observed that all three spectra reduced to the magnetic Compton profile, describing the spin-dependent ground-state momentum density, and that within the experimental error the integrated intensity of the magnetic effect scaled as predicted by the cross section derived in the limit of energies much less than the rest energy of the electron. The magnetic Compton profile of Fe [111], measured using 167.2 keV incident energy and with momentum resolution of 0.42 a.u., was compared with the prediction from a full-potential linearized augmented plane-wave model profile. The fine structure predicted by theory was confirmed by the experimental profile at this improved resolution. PMID- 16699215 TI - Brilliance--an overview. AB - One of the most important figures of merit for third-generation sources is the brilliance. An intuitive physical picture of the diffraction properties of synchrotron radiation is presented in this paper. The consequences of these diffraction properties on the design of third-generation light sources is also discussed. PMID- 16699216 TI - Status of MAX II. AB - A new third-generation VUV synchrotron radiation ring called MAX II has just been built and commissioned in Lund, Sweden. Owing to tight boundary conditions regarding economy, space and manpower some novel techniques have been used in the design of the ring. The present status of the ring as well as the motivation for the parameter choices and techniques chosen are presented. PMID- 16699217 TI - X-ray optics developments at the APS for the third generation of high-energy synchrotron radiation sources. AB - Third-generation hard-X-ray synchrotron radiation sources simultaneously provide both a need and an opportunity for the development of new short-wavelength optical components. The high power and power densities of the insertion-device produced X-ray beams have forced researchers to consider what may seem like exotic approaches, such as cryogenically cooled silicon and highly perfect diamond crystals, to mitigate thermal distortions in the first optical components. Once the power has been successfully filtered while maintaining the high beam brilliance, additional specialized optical components can be inserted into the monochromatic beam that take advantage of that brilliance. This paper reviews the performance of such optical components that have been designed, fabricated and tested at the Advanced Photon Source, starting with high-heat-load components and followed by examples of several specialized devices, such as an meV resolution (in-line) monochromator, a high-energy X-ray phase retarder and a phase-zone plate with submicrometer focusing capability. PMID- 16699218 TI - Asymmetric Fraunhofer diffraction from roller-blade slits. AB - X-ray diffraction experiments under coherent conditions have been made possible by the development of new sources of synchrotron radiation, but make tough demands on the experimental apparatus. Here we describe the design and initial testing of a precision aperture that uses polished molybdenum rods as the slit blades. The device has an inherent asymmetry which is accurately accounted for by a simple modification to the Fraunhofer diffraction function. PMID- 16699219 TI - A system for integrated collection and analysis of crystallographic diffraction data. AB - A set of linked software modules has been installed at the A1 station of CHESS, the Cornell High Energy Synchrotron Source, which, with the underlying hardware, allows crystallographic users of the facility to evaluate crystals, collect diffraction images and process the images rapidly and with assurance of the quality of the resultant data, thereby making most efficient use of their beam time. The system includes a CCD detector and its controlling software, with a graphical user interface, a convenient oscillation camera featuring automated alignment with the X-ray beam, a program for optimizing crystal rotation range, and the HKL data-reduction package. Principles embodied in this system are applicable to other facilities where crystallographic data are routinely collected. PMID- 16699220 TI - Large-format imaging plate and weissenberg camera for accurate protein crystallographic data collection using synchrotron radiation. AB - Off-line and on-line protein data-collection systems using an imaging plate as a detector are described and their components reported. The off-line scanner IPR4080 was developed for a large-format imaging plate ;BASIII' of dimensions 400 x 400 mm and 400 x 800 mm. The characteristics of this scanner are a dynamic range of 10(5) photons pixel(-1), low background noise and high sensitivity. A means of reducing electronic noise and a method for finding the origin of the noise are discussed in detail. A dedicated screenless Weissenberg camera matching IPR4080 with synchrotron radiation was developed and installed on beamline BL6B at the Photon Factory. This camera can attach one or two sheets of 400 x 800 mm large-format imaging plate inside the film cassette by evacuation. The positional reproducibility of the imaging plate on the cassette is so good that the data can be processed by batch job. Data of 93% completeness up to 1.6 A resolution were collected on a single axis rotation and the value of R(merge) becomes 4% from a tetragonal lysozyme crystal using a set of two imaging-plate sheets. Comparing two types of imaging plates, the signal-to-noise ratio of the ST-VIP-type imaging plate is 25% better than that of the BASIII-type imaging plate for protein data collection using 1.0 and 0.7 A X-rays. A new on-line protein data-collection system with imaging plates is specially designed to use synchrotron radiation X rays at maximum efficiency. PMID- 16699221 TI - Applications of high-energy synchrotron radiation for structural studies of polycrystalline materials. AB - The large penetration power of high-energy X-rays (>60 keV) raises interesting prospects for new types of structural characterizations of polycrystalline materials. It becomes possible in a non-destructive manner to perform local studies, within the bulk of the material, of the fundamental materials physics properties: grain orientations, strain, dislocation densities etc. In favourable cases these properties may be mapped in three dimensions with a spatial resolution that matches the dimensions of the individual grains. Imbedded volumes and interfaces become accessible. Moreover, the high energies allow better in situ studies of samples in complicated environments (industrial process optimization). General techniques for research in this energy range have been developed using broad-band angle-dispersive methods, on-line two-dimensional detectors and conical slits. Characterizations have been made at the level of the individual grains and grain boundaries as well as on ensembles of grains. The spatial resolution is presently of the order of 10-100 micom. Four examples of applications are presented along with an outlook. PMID- 16699222 TI - Compression behaviour of elastically anisotropic polycrystals using energy dispersive X-ray diffraction. AB - The compression behaviour of a foil of Cu3Au in a pressure medium of NaCl has been studied by energy-dispersive X-ray diffraction in a diamond-anvil cell. Evidence from stress analysis and peak broadening of the foil (the most extreme example of a non-ideal powder) throws light on the compression of powders under non-hydrostatic conditions. A complete pressure cycle, including re pressurization after pressure release, shows that significant plastic deformation takes place which results in large deviations from a (hydrostatic) equation of state. The origin of the deformations is traced to shear stresses transmitted to the sample through the pressure medium. PMID- 16699223 TI - Synchrotron radiation study of the relation between structure and strain in polyurethane elastomers. AB - This paper describes a system for the study of the relation between structure and applied strain in thermoplastic polyurethane elastomers using the Australian National Beamline Facility at the Photon Factory, KEK, Tsukuba, Japan. The system uses the sagittal focusing monochromator at beamline 20B to provide a high intensity focused beam which then falls on the specimen mounted in a miniature tensometer mounted in the unique vacuum diffractometer (BIGDIFF). Imaging plates were used to record simultaneously SAXS and WAXS patterns from the specimen at a particular strain. The change in SAXS and WAXS patterns with loading and unloading was recorded using a ten-plate imaging-plate changer. PMID- 16699224 TI - Application of imaging-grazing-incidence X-ray diffraction and specular reflectivity to the structural investigation of quantum-confinement semiconductor devices. AB - X-ray reflectivity and grazing-incidence X-ray diffraction techniques have been employed to investigate the structure of quantum-well and quantum-dot semiconductor devices. This work has been performed using both laboratory and synchrotron radiation sources. The use of synchrotron radiation enabled reflectivity studies to be performed on small samples, and established the feasibility of imaging-grazing-incidence diffraction studies on quantum confinement structures. Interdiffusion effects in quantum-well and quantum-dot structures, the disordering of overlayers grown on quantum dots, and the variation in diffraction pattern with incident angle have been observed. It is evident that X-ray reflectivity and imaging-grazing-incidence X-ray diffraction yield quite different but complementary information. PMID- 16699225 TI - X-ray reflectivity at the L edges of Gd. AB - Preparations are underway for the experimental investigation of the roughness of magnetic interfaces in rare-earth multilayers by combining the grazing-angle X ray scattering technique with the resonant magnetic scattering of hard X-rays. Theoretical considerations show that for small scattering angles, 2theta, the asymmetry ratio, A = [I(+) - I(-)]/[I(+) + I(-)], depends on 2theta and varies as 1/cos theta. The first step towards the goal of determining the magnetic roughness has been taken by measuring the chemical roughness (via specular reflectivity) of a Gd thin-film sample at five photon energies close to the L(3) absorption edge, which yielded the dispersion corrections, f' and f'', to the Gd atomic form factor in good agreement with the calculation of Cromer & Liberman [J. Chem. Phys. (1970), 53, 1891-1898]. PMID- 16699226 TI - On the off-Bragg reflectivity enhancement in dynamical multiple diffraction of X rays by an In(0.5)Ga(0.5)P/GaAs heteroepitaxial structure. AB - The effect of dynamical interactions between reflections involved in 000/111/220 multiple diffraction by an In(0.5)Ga(0.5)P/GaAs heteroepitaxial structure was observed, which significantly increases the intensity of the (111) reflection in the region of the total (220) reflection from the substrate during Renninger-like scanning. The amplitude of the peak is roughly five times as great as the peak of detoured excitation for ideal GaAs. Possible applications of the observed feature in synchrotron-radiation-based studies are discussed. PMID- 16699227 TI - X-ray reflectivity study of semiconductor interfaces. AB - The results of an X-ray reflectivity study of thick AlAs-AlGaAs and thin Ge-Si-Ge multilayers grown using metal-organic vapour-phase epitaxy and ion-beam sputtering deposition techniques, respectively, are presented. Asymmetry in interfaces is observed in both of these semiconductor multilayers. It is also observed that although the Si-on-Ge interface is sharp, an Si(0.4)Ge(0.6) alloy is formed at the Ge-on-Si interface. In the case of the III-V semiconductor, the AlAs-on-AlGaAs interface shows much greater roughness than that observed in the AlGaAs-on-AlAs interface. For thin multilayers it is demonstrated that the compositional profile as a function of depth can be obtained directly from the X ray reflectivity data. PMID- 16699229 TI - A novel insertion device for circularly polarized radiation. AB - A novel insertion device for circularly polarized radiation with a wide available energy range is proposed. It consists of two helical undulators with different period lengths. From an analysis made on the radiation, higher harmonics are found to be contained in the radiation, unlike the ordinary helical undulator. In addition, calculations show that a high degree of circular polarization is obtained for finite beam emittance. PMID- 16699230 TI - Two-photon correlations in X-rays from a synchrotron radiation source. AB - The intensity correlation experiment using visible light performed by Hanbury Brown & Twiss [Nature (London) (1956), 177, 27-29] is extended to X-ray wavelengths. Correlations of 14.4 keV X-ray photons from a synchrotron radiation source are observed by the coincidence counting technique. The high brilliance of synchrotron radiation available in the Tristan main ring enables the observation of two-photon correlations with a reasonable measurement time. PMID- 16699231 TI - Diamond beam-position monitor for undulator radiation and tests at the Tristan Super Light Facility. AB - A photon beam-position monitor using a diamond foil as a position-sensitive device has been developed for use on insertion-device beamlines of third generation synchrotron radiation facilities such as SPring-8, and was tested on the undulator beamline of the Tristan Super Light Facility at KEK. The beam test results show that the diamond monitor can be operated in a photoconductive mode like a semiconductor detector. It has a linear working range of about +/-1 mm and a position sensitivity of less than 3 mum. The stability of the monitor was confirmed by continuous operation under low photon beam intensity conditions. PMID- 16699232 TI - Slits as adjustable pinholes for coherent X-ray scattering experiments. AB - The combination of accurate translation stages with carefully polished slit blades leads to slits that have many advantages as pinholes for coherent X-ray scattering experiments. The size is adjustable and can be made as small as 0.5 mum. Setting up is easy, while the blade thickness (1 mm tungsten) also makes the slits useful for hard X-rays. A relation between the slit-sample distance and the minimum beam size, together with the corresponding slit size, is derived. This shows that a micrometer-sized beam can be achieved with this type of slits. PMID- 16699233 TI - Application of energy-resolved measurements to Laue diffraction: determination of unit-cell parameters, deconvolution of harmonics and assignment of systematic absences. AB - The use of energy-resolved area detection of Laue diffraction patterns for the determination of unit-cell parameters and systematic absences is demonstrated. Seven different crystals having previously known unit cells were re-examined using Laue diffraction methods. These crystals included four different crystal systems including cubic, orthorhombic, tetragonal and monoclinic cells. The crystals had cell sizes ranging from 179.4 to 4588.3 A(3). Comparison of known and re-determined cells showed good agreement (ratio of known to measured cells = 0.987 +/- 0.18). A single procedure was suitable for all unit-cell determinations. The accuracy of the method is presently limited by the quality of the available energy measurements. Some of the crystals represent space groups containing systematic absences normally obscured by harmonic overlap when using the Laue method. These include absences due to 2(1) screw axes (h, k or 1 = 2n + 1) and cell centering (h + k = 2n + 1). All systematic absences were identified using a combination of multiple linear regression with either stepwise elimination or stepwise inclusion and an F test for assignment of systematic absence. The methods are discussed in detail and simulations are used to evaluate critical tolerances for future systems. PMID- 16699234 TI - Investigation of the variation in orientation and crystallinity in poly(ethylene terephthalate) containers using microfocus X-ray diffraction. AB - The microfocus X-ray beamline at the European Synchrotron Radiation Facility has been used to investigate the variation in molecular orientation and crystallinity in the wall of a container fabricated from poly(ethylene terephthalate). Two dimensional wide-angle X-ray scattering patterns were recorded and displayed in real time as the specimen was tracked across the incident X-ray beam enabling the measurement of textural changes to be made with a spatial resolution of ~2 mum. PMID- 16699235 TI - The use of a scanning X-ray microprobe for simultaneous XRF/XRD studies of fly ash particles. AB - With the opening of the first real ;third-generation' synchrotron source in Grenoble, in fall 1994, X-ray sources of unprecedented brilliances and qualities became available to the scientific community. Different X-ray analytical techniques could now be applied on a level that was unimaginable only a decade ago. Here are some preliminary results from an experiment where different analytical techniques have been applied on a micrometer level carried out at the most powerful synchrotron microbeam currently available in the world, the microfocus beamline (BL1) at ESRF. This beamline can now provide micrometer-sized X-ray beams with a flux density up to 10(10) photons microm(-2) at an energy of 13 keV and with a bandwidth of 10(-4). In this experiment, X-ray diffraction and X-ray fluorescence have been combined in order to obtain a precise and comprehensive micro-analytical description of micrometer-sized fly ash particles. These types of particles are heavily inhomogeneous with a very irregular shape that makes them inaccessible to conventional micro-analysis. The experiment was performed in a scanning mode and two-dimensional images of different analytical information were reconstructed from the data recorded during the scan. The major features and limitations of this micro-analytical technique will be outlined and different examples on how the analytical information can be used for generating two-dimensional images of the sample will be demonstrated and discussed. PMID- 16699236 TI - High-resolution X-ray fluorescence and excitation spectroscopy of metalloproteins. AB - A spectrograph has been developed with sufficient efficiency to make high resolution fluorescence experiments on metalloproteins possible. The resolution of this spectrometer can reach 0.45 eV at 7.1 keV emission energy. The focus images of this multiple curved-crystal array spectrometer are presented. The chemical sensitivity of Kbeta emission spectra can be used to identify chemical states, and the spin-polarized near-edge structure provides a new measure of the spin density. The high-resolution fluorescence metalloprotein studies should become routine with third-generation synchrotron facilities, and the strength of both site and spin selectivity should complement the structural information from other spectroscopies. PMID- 16699237 TI - EXAFS Debye-Waller factor and thermal vibrations of crystals. AB - The EXAFS Debye-Waller factor depends on the correlation of atomic motion and can yield original information on the vibrational dynamics of crystalline solids. In this paper an introductory treatment of thermal disorder in EXAFS, based on thc cumulant approach, is given. Thc general relation between mean-square relative displacement (MSRD) measured by EXAFS and atomic thermal vibrations in harmonic approximation is explored. Strengths and limitations of the phenomenological Einstein- and Debye-correlated models are discussed. Some of the most significant results so far obtained are reviewed. A relatively simple method for estimating anharmonic corrections to the MSRD is proposed. PMID- 16699238 TI - Relaxation process of the excited state and selection rule in the soft X-ray emission of Si and cBN. AB - The soft X-ray emission spectra of Si and cubic boron nitride (cBN) are compared in the Si2p- and B1s-core exciton regions because inversion symmetry is lost and the band gap is extremely large for cBN although both crystal structures are very similar. The background in the soft X-ray emission spectrum of Si is much larger than that for cBN but becomes extremely weak as the excitation energy becomes low and the Raman process becomes dominant compared with the fluorescence process. The background is found to be mainly formed by electron excitation from the valence band to the conduction band. The Raman spectrum is similar to the absorption spectrum in cBN, while those of Si are quite different from each other. It is shown that the centrosymmetry is important in the selection rule of Raman scattering. In resonant Raman scattering the selection rule partly breaks down due to the lattice relaxation process that emits an ungerade phonon in the intermediate state. PMID- 16699239 TI - Gauss-schell sources as models for synchrotron radiation. AB - Gauss-Schell light sources are considered as models for synchrotron radiation. These sources can be viewed as random superpositions of coherent Gaussian beams. The relationships of the various widths that can be defined for the description of intensity and coherence, as related to the widths of the electron beam and the single-electron radiation (diffraction) pattern, are summarized. The description is also applied to the temporal coherence, which is of interest in the case of free-electron lasers. PMID- 16699240 TI - A Two-Dimensional X-ray Scattering System for In-Situ Time-Resolving Studies of Polymer Structures Subjected to Controlled Deformations. AB - A two-dimensional X-ray scattering system developed around a CCD-based area detector is presented, both in terms of hardware employed and software designed and developed. An essential feature is the integration of hardware and software, detection and sample environment control which enables time-resolving in-situ wide-angle X-ray scattering measurements of global structural and orientational parameters of polymeric systems subjected to a variety of controlled external fields. The development and operation of a number of rheometers purpose-built for the application of such fields are described. Examples of the use of this system in monitoring degrees of shear-induced orientation in liquid-crystalline systems and crystallization of linear polymers subsequent to shear flow are presented. PMID- 16699241 TI - A New High-Flux Chemical and Materials Crystallography Station at the SRS Daresbury. 1. Design, Construction and Test Results. AB - A new single-crystal diffraction facility has been constructed on beamline 9 of the SRS at Daresbury Laboratory for the study of structural problems in chemistry and materials science. The station utilizes up to 3.8 mrad horizontally from the 5 T wiggler magnet which can be focused horizontally and vertically. The horizontal focusing is provided by a choice of gallium-cooled triangular bent Si (111) or Si (220) monochromators, giving a wavelength range from 0.3 to 1.5 A. Focusing in the vertical plane is achieved by a cylindrically bent zerodur mirror with a 300 mum-thick palladium coating. The station is equipped with a modified Enraf-Nonius CAD-4 four-circle diffractometer and a Siemens SMART CCD area detector system. High- and low-temperature facilities are available to cover the temperature range from about 80 to 1000 K. Early results on test compounds without optimization of the beam optics demonstrate that excellent refined structures can be obtained from samples giving diffraction patterns too weak to be measured with conventional laboratory X-ray sources, fulfilling a major objective of the project. PMID- 16699242 TI - Protein Crystallography at Ultra-Short Wavelengths: Feasibility Study of Anomalous-Dispersion Experiments at the Xenon K-edge. AB - A protein crystallography experiment at the xenon K-edge (lambda = 0.358 A) has been successfully carried out at the materials science beamline (BL2/ID11) of the ESRF. The samples used in this methodological study were crystals of porcine pancreatic elastase, a 26 kDa protein of known structure. The diffraction data are of excellent quality. The combination of isomorphous replacement and anomalous dispersion of a single xenon heavy-atom derivative allowed accurate phase determination and the computation of a high-quality electron density map of the protein molecule. This is the first fully documented report on a complete protein crystallography experiment, from data collection up to phase determination and calculation of an electron density map, carried out with data obtained at ultra-short wavelengths. Experimental considerations as well as possible advantages and drawbacks of protein crystallography at very short and ultra-short wavelengths are discussed. PMID- 16699243 TI - Anomalous Dispersion with Edges in the Soft X-ray Region: First Results of Diffraction from Single Crystals of Trypsin Near the K-Absorption Edge of Sulfur. AB - Anomalous dispersion of X-ray diffraction at wavelengths near the X-ray K absorption edge of sulfur at wavelengths around 5 A has been applied to single crystals of trypsin obtained from an ammonium sulfate solution. The multiwavelength anomalous-dispersion method based on 775 unique reflections (+183 Bijvoet mates) measured at three wavelengths near the K-absorption edge of sulfur in trypsin (two methionines and disulfide bridges of six cystines) reproduces the known features of the trypsin structure of a resolution of 4 A. It appears that there is anisotropic anomalous scattering from the disulfide bridges of cystine. The multiwavelength anomalous solvent contrast shows up at wavelengths near the K absorption edge of the sulfate ions, which is shifted by 10 eV to higher energies with respect to that of sulfur in trypsin. The influence of the complex contrast of trypsin in 2.5 M ammonium sulfate on the dispersion of a low-order reflection is analyzed. The measurement of anomalous dispersion of X-ray diffraction at long wavelengths beyond 5 A requires a special diffractometer, the features of which are presented. An outstanding one is a detector system consisting of four multiwire proportional counters. Its efficiency is compared with that of imaging plates. The influence of radiation damage with soft X-ray diffraction from single crystals of trypsin is presented and possible remedies are discussed. PMID- 16699244 TI - Operation of a Two-Crystal X-ray Interferometer at the Photon Factory. AB - A two-crystal X-ray interferometer for phase-contrast X-ray imaging is reported. Mechanical stability of less than 0.1 nm is required to operate the two-crystal interferometer. The feasibility of using such an interferometer at the Photon Factory using synchrotron radiation has been investigated. Interference fringes of 70% visibility were observed with 0.092 nm X-rays. This result indicates that the two-crystal X-ray interferometer can be applied to phase-contrast X-ray imaging. PMID- 16699245 TI - New commercial products. PMID- 16699246 TI - Fifty years of synchrotron radiation. PMID- 16699247 TI - Atomic and molecular physics using synchrotron radiation - the early years. AB - The field of atomic and molecular photoionization has undoubtedly been transformed by the use of synchrotron radiation. In the mid-1960's the 180 MeV electron synchrotron at the National Bureau of Standards, Washington, DC, was used to discover many resonances in the photoionization continua of atoms and simple molecules. The resonances were characterized in terms of the excited states involved, their energies and lifetimes. This article concentrates on that period. As dedicated high-flux storage rings became available, the emphasis turned to the study of the products of the photoionization process in order to further understand the electron correlation effects that had been uncovered earlier; examples of these more sophisticated experiments are given. PMID- 16699248 TI - Tantalus, a 240MeV Dedicated Source of Synchrotron Radiation, 1968-1986. AB - Tantalus was a 240 MeV electron storage ring completed and commissioned in 1968. It was the first storage ring operated exclusively for the production of synchrotron radiation, although it was not designed for that purpose. As such, it influenced the operating pattern for subsequent dedicated sources of synchrotron radiation. Pioneering experiments using synchrotron radiation were carried out on this machine, and innovative instrumentation was produced there. It ceased operation in 1986. PMID- 16699249 TI - Synchrotron Radiation Research in the UK. AB - The origins, development and growth of synchrotron radiation science and technology in the UK, and an account of the expansion of the synchrotron radiation research programme, its highlights, current activities and the prospects for the future expansion of synchrotron radiation activities within the UK, are presented. PMID- 16699250 TI - A prospect and retrospect - the Japanese case. AB - The early through recent history of synchrotron radiation research in Japan, since the initial efforts in 1962, is reviewed. Following a period of parasitic use of an electron synchrotron, Japanese users attempted to build a storage ring as a dedicated soft X-ray source, which was completed in 1974. It opened up a new era of second-generation synchrotron radiation research. The Photon Factory, a dedicated X-ray source commissioned in 1982, provided a much wider research area as well as a number of technical innovations, among which insertion devices brought the further prospect of significant improvements in the properties of sources. As a consequence, the new concept of a light source oriented towards full exploitation of insertion devices, or the idea of a third-generation source, was created. The motivations and developments which led to Spring-8, a third generation Japanese X-ray source that is currently being commissioned, will be reviewed briefly. PMID- 16699251 TI - Development of Synchrotron Radiation as a High-Intensity Source for X-ray Diffraction. AB - Interest in the molecular mechanism of muscle contraction led to the search for an intense source of X-rays of 1-2 A wavelength so as to be able to examine the rich X-ray diffraction patterns given by muscles during contraction. This led to the first X-ray diffraction experiments using synchrotron radiation, carried out by Holmes, Rosenbaum and Witz at DESY, Hamburg, in September 1970. In the following years, the EMBL Outstation, to utilize synchrotron radiation for biological structure determination, was established at DESY and preliminary experiments on muscle were also carried out at NINA (Daresbury). The development of time-resolved techniques for muscle diffraction was first started in the MRC Molecular Biology Laboratory in Cambridge, using rotating-anode X-ray tubes, and was then greatly extended at the EMBL Outstation, Hamburg, using the storage ring DORIS. This was a very successful venture, and helped to drive the whole technology development and to interest other potential users in the technique. PMID- 16699252 TI - Early work with synchrotron radiation at stanford. AB - The use of synchrotron radiation in the soft and hard X-ray spectral region received major impetus with the start of parasitic operation of the Stanford Synchrotron Radiation Project (SSRP) in 1974. This was the first time that synchrotron radiation from a multi-GeV electron storage ring was made available in a user facility for studying the structure of matter. Here we review the early work at SSRP as well as the activities that preceded it, highlighting the scientific accomplishments (soft X-ray photoemission, EXAFS, protein crystallography), beamline instrumentation developments and source improvements. The early work using bending-magnet radiation led to the funding of several dedicated facilities in the US and elsewhere in the world - the so-called second generation light sources. Early work with wiggler and undulator insertion devices led to funding of third-generation sources better optimized for insertion device sources, particularly undulators. PMID- 16699253 TI - Prices of IUCr journals. PMID- 16699254 TI - Long-term bosentan in chronic thromboembolic pulmonary hypertension. AB - BACKGROUND: There is no approved pharmacological treatment for patients with chronic thromboembolic pulmonary hypertension (CTEPH) who are not suitable for pulmonary endarterectomy (PEA). OBJECTIVE: The study investigates the effect of the dual endothelin receptor antagonist bosentan on exercise tolerance (6-min walking distance, 6MWD) and right ventricular function (Tei index) in patients with CTEPH over 24 months. METHODS: Twelve consecutive patients (5 males and 7 females) with CTEPH not eligible for PEA or following partial or complete failure of PEA were included in a non-randomized, open-label prospective study. All patients were WHO class III. They were included, if progressive pulmonary hypertension was diagnosed despite best supportive treatment. Bosentan was started at 62.5 mg b.i.d. and increased to the final dose of 125 mg b.i.d. RESULTS: 6MWD and the Tei index were assessed every 6 months. We observed a significant increase in 6MWD from 319 +/- 85.0 m at baseline to 391 +/- 76.9 m at 6 months and a significant decrease in the Tei index from 0.39 +/- 0.10 at baseline to 0.34 +/- 0.08 at 6 months. This improvement was maintained over 24 months (6 MWD: 381 +/- 101 m; Tei index: 0.31 +/- 0.03). Six patients exhibited an improvement in WHO class at 6, 12 and 18 months, 5 demonstrated improvement at the 24-month follow-up. The remainder were stable throughout the study period. CONCLUSION: This is the first study demonstrating a long-term beneficial effect of bosentan on exercise tolerance (6MWD) and right heart function (Tei index) in CTEPH. PMID- 16699255 TI - A case of severe dyspnea and an unusual bronchoscopy: the Chilaiditi syndrome. PMID- 16699256 TI - Characteristics of multiple primary malignancies associated with lung cancer by gender. AB - BACKGROUND: There are gender differences in multiple primary malignancies associated with lung cancer (MPMLC) in terms of clinical characteristics. However, the importance of these differences in the management of patients has not been clarified. OBJECTIVE: Differences in characteristics affected by gender were investigated in MPMLC to identify factors important for the proper management of the patients. METHODS: Univariate and multivariate analyses were performed between 82 male and 34 female patients with MPMLC treated from August 1982 to March 2002. RESULTS: In univariate analysis, the numbers of smokers or ex smokers, smoking-related cancer and synchronous multiple primary malignancies were significantly increased in males with MPMLC (p < 0.0001, p < 0.05 and p < 0.05, respectively). In multivariate analysis, synchronous multiple primary malignancies and the number of smokers or ex-smokers were significantly different between male and female MPMLC. Gastric, lung and colon cancers were major constituents in male MPMLC, and 40.2% of all malignancies were smoking-related cancers. On the other hand, breast and uterine cancers were major constituents in female MPMLC, and only 20.6% of all MPMLC were smoking-related cancers. CONCLUSIONS: Male patients with MPMLC demonstrated significant smoking history and synchronous multiple primary malignancies, indicating the need for different approaches to properly manage and follow up male versus female MPMLC patients. PMID- 16699258 TI - Active influenza immunization in hemodialysis patients: comparison between single dose and booster vaccination. AB - BACKGROUND/AIMS: Hemodialysis (HD) patients are recommended to be immunized against influenza annually, but the immune response after vaccination is known to be weakened in these patients. We intended to compare the efficacy of influenza vaccines in HD patients with that in healthy people, and we also evaluated the additive effect of a booster vaccination in HD patients. METHODS: During the 2003 2004 influenza season, 100 patients on HD and 50 age-matched healthy controls were recruited from the Korea University Guro Hospital. The HD patients were divided into two groups: single-dose group (50 patients) and booster group (50 patients). Eight weeks following the influenza vaccination, the serum antibody responses were compared. RESULTS: Although the antibody response of the HD patients was impaired in comparison with that of healthy controls, more than 90% of the HD patients showed protective antibody levels. Booster vaccination did not significantly increase the immune response in HD patients. CONCLUSIONS: Influenza vaccination in HD patients should be encouraged, but the immune response was comparatively impaired in subpopulations, including patients with a long-term HD history (>2.5 years), with a higher urea reduction rate, or with underlying diabetes. There was no effect of a booster vaccine dose on the antibody titers. PMID- 16699257 TI - Regulation of acid-base transporters by vasopressin in the kidney collecting duct of Brattleboro rat. AB - AIM: The objective of these studies was to examine the effects of long-term vasopressin treatment on acid-base transporters in the collecting duct of rat kidney. METHODS: Brattleboro rats were placed in metabolic cages and treated with daily injections of 1-desamino-8-D-arginine vasopressin (dDAVP), a selective V2 receptor agonist, or its vehicle (control) for up to 8 days. RESULTS: dDAVP treatment resulted in a significant reduction in serum bicarbonate concentration, and caused the upregulation of key ammoniagenesis enzymes, along with increased urinary NH4+ excretion. Northern hybridization and immunofluorescence labeling indicated a significant increase (+80%) in mRNA expression of the apical Cl-/HCO3 exchanger pendrin (PDS), along with a sharp increase in its protein abundance in B-type intercalated cells in the cortical collecting duct in dDAVP-treated rats. In the inner medullary collecting duct, the abundance of basolateral Cl-/HCO3- exchanger (AE1) and apical H+-ATPase was significantly reduced in dDAVP-treated rats. Kidney renin mRNA increased significantly and correlated with an increase in serum aldosterone levels in dDAVP-injected rats. Serum corticosterone levels were, however, reduced and correlated with increased mRNA levels of renal 11beta hydroxysteroid dehydrogenase-2 (11beta-HSD2) and decreased mRNA expression of 11beta-hydroxylase in the adrenal gland of dDAVP-injected rats. CONCLUSION: Chronic administration of dDAVP to Brattleboro rats is associated with the upregulation of PDS and downregulation of H+-ATPase and AE1 in the collecting duct, along with increased ammoniagenesis. Stimulation of the renin-angiotensin aldosterone system and/or decreased glucocorticoid levels likely plays a role in the transduction of these effects. PMID- 16699259 TI - Speech recognition at 1-year follow-up in the childhood development after cochlear implantation study: Methods and preliminary findings. AB - The Childhood Development after Cochlear Implantation (CDaCI) study is a longitudinal multicenter investigation designed to identify factors influencing spoken language in young deaf children with cochlear implants. Normal-hearing peers serve as controls. As part of a comprehensive evaluation battery, a speech recognition hierarchy was designed to assess how well these children recognize speech stimuli across developmental stages. Data were analyzed for the earliest measures in 42 pairs of children reaching 1 year of follow-up. A number of children in the cochlear implant group who met criteria for testing approached levels of performance similar to the normal-hearing controls, and some could identify sentences in competition. These results demonstrate the responsiveness of the speech recognition hierarchy in tracking emergent skills from a sample of the CDaCI cohort. PMID- 16699260 TI - Relevance of tenascin-C and matrix metalloproteinases in vascular abnormalities in murine hypoplastic lungs. AB - BACKGROUND: Tenascin-C (TN-C), an extracellular matrix glycoprotein, is crucial to cell-migration, proliferation, apoptosis and remodeling of tissues, with a potential role in pathobiology of pulmonary hypertension. Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) are crucial to the integrity of the extracellular matrix. TN-C and MMPs are counter regulatory molecules, which influence the vascular integrity through modulations of elastin. We have a murine model of pulmonary hypoplasia with coexistent diaphragmatic hernia, vascular abnormalities and excessive arterial smooth muscle cell (SMC) proliferation. OBJECTIVES: Our objective was to investigate modulations of TN-C and MMPs in hypoplastic lungs and their possible contribution to the observed pulmonary vascular abnormalities. METHODS: We addressed our objectives by pursing immunoblotting and immunohistochemistry and zymography/reverse zymography to assess the alterations in activities of MMPs and their inhibitors. RESULTS: We observed significant down-regulation of MMP-9 activity in hypoplastic lungs at the later fetal developmental stages, whereas MMP-2 activity assessed by gelatin zymography remained unaltered. Reverse zymography revealed up-regulation of activities of TIMP-1, -2, -3 and -4 in hypoplastic lungs during later fetal development, with pronounced increases in TIMP-3 and -4 activities. Furthermore, immunoblot analyses and immunohistochemistry revealed that TN-C protein was down-regulated in developing hypoplastic lungs, compared to normal lungs. CONCLUSIONS: (1)TN-C is known to inhibit vascular SMC proliferation. But, decrease in TN-C in hypoplastic lungs may support the observed arterial SMC proliferation. (2) Our studies showed that in hypoplastic lungs the SMC apoptosis is not affected, thus suggesting that SMC proliferation and apoptosis may be two separate processes in pulmonary hypoplasia with coexistent diaphragmatic hernia. Together, our data showed an imbalance in the extracellular matrix proteins, which may contribute to the pulmonary vascular abnormalities. PMID- 16699261 TI - Advances in GI pharmacology and therapeutics. PMID- 16699262 TI - Acid suppression therapy: where do we go from here? AB - The dramatic success of pharmacological acid suppression in healing peptic ulcers and managing patients with gastroesophageal reflux disease (GERD) has been reflected in the virtual abolition of elective surgery for ulcer disease, a reduction in nonsteroidal anti-inflammatory drug (NSAID)-associated gastropathy and the decision by most patients with reflux symptoms to continue medical therapy rather than undergo surgical intervention. However, a number of challenges remain in the management of acid-related disorders. These include management of patients with gastroesophageal symptoms who do not respond adequately to proton pump inhibitor (PPI) therapy, treatment of patients with nonvariceal upper gastrointestinal bleeding, prevention of stress-related mucosal bleeding, optimal treatment and prevention of NSAID-related gastrointestinal injury, and optimal combination of antisecretory and antibiotic therapy for the eradication of Helicobacter pylori infection. A number of new drugs are currently being investigated to provide a significant advance on current treatments. Some of them (namely potassium-competitive acid blockers (P-CABs) and CCK2-receptor antagonists) have already reached clinical testing while some others (like the antigastrin vaccine, H3-receptor ligands or gastrin-releasing peptide receptor antagonists) are still in preclinical development and need the proof of concept in human beings. Of the current approaches to reduce acid secretion, P-CABs and CCK2-receptor antagonists hold the greatest promise, with several compounds already in clinical trials. Although the quick onset of action of P-CABs (i.e. a full effect from the first dose) is appealing, the results of phase II studies with one such agent (namely AZD0865) did not show any advantages over esomeprazole. Thanks to their limited efficacy and the development of tolerance it is unlikely that CCK2 antagonists will be used alone as antisecretory compounds but, rather, their combination with PPIs will be attempted with the aim of reducing the long-term consequences of hypergastrinemia. While H2-receptor antagonists (especially soluble or over-the-counter formulations) will become the 'antacids of the third millennium' and will be particularly useful for on-demand symptom relief, clinicians will continue to rely on PPIs to control acid secretion in GERD and other acid-related diseases. In this connection, several new PPI formulations have been developed and two novel drugs (namely ilaprazole and tenatoprazole) are being studied in humans. The recently introduced immediate release (IR) omeprazole formulation (currently available only in the USA) quickly increases intragastric pH and, given at bedtime, seems to achieve a better control of nocturnal acidity. IR formulations of other PPIs (including the investigational ones) will probably be available in the future and will enlarge our therapeutic armamentarium. Amongst the novel PPIs, tenatoprazole appears to be a true advance in the acid suppression therapy. Its long half-life (the longest among the available compounds) and longer duration of antisecretory action, with no difference between day and night, will allow the drug to go beyond the intrinsic limitations of currently available PPIs. Thanks to its favorable pharmacokinetics, the sodium salt of S-tenatoprazole is being developed and the preliminary results indicate that this drug has the potential to address unmet clinical needs. Although some decades have elapsed since the introduction of effective and safe antisecretory drugs in clinical practice and their use has stood the test of time, the ongoing research will further provide the clinician with more effective means of controlling acid secretion. PMID- 16699263 TI - Antisecretory drugs for diarrheal disease. AB - Acute diarrhea is a major cause of morbidity and mortality worldwide. Infants and pre-school children are the most vulnerable in whom there are 2-3 million deaths each year as a result of the associated dehydration and acidosis. Although oral rehydration therapy has reduced mortality during the past 30 years ago, the search for agents that will directly inhibit intestinal secretory mechanisms and thereby reduce faecal losses in patients with high-volume watery diarrhea has continued for more than 20 years. A variety of potential targets for antisecretory agents have been explored which include loci within the enterocyte (the chloride channel, calcium-calmodulin) and other sites such as enteric nerves and endogenous mediators (such as 5-HT, prostaglandins). Although the potential of calcium-calmodulin inhibition has as yet not been realised, preliminary studies suggest that there are chloride channel blockers under development that will find a place in the management of secretory diarrheas. Recent work has highlighted the importance of neurohumoral mechanisms in the pathogenesis of acute diarrhea. Potentiation of the effects of endogenous enkephalin activity by enkephalinase inhibition has already produced a safe, effective anti-secretory drug, racecadotril. Speculative early work indicates that there may be a role for antagonists of 5-HT, substance P, and VIP receptors. There now seems to be a real possibility that antisecretory therapy will become more widely available in the future. PMID- 16699264 TI - Drugs acting on serotonin receptors for the treatment of functional GI disorders. AB - In the gut, serotonin (5-hydroxytryptamine: 5-HT) exerts a variety of effects on intrinsic enteric neurons, extrinsic afferents, enterocytes and smooth muscle cells, which are related to the expression of multiple 5-HT receptor types and subtypes regulating motility, vascular tone, secretion and perception. Agonists and antagonists at 5-HT receptors have gained access to the market for the two major variants of the irritable bowel syndrome (IBS), a functional disorder characterized by abdominal pain associated with diarrhea and/or constipation in the absence of any organic abnormality. Indeed, the 5-HT3 receptor antagonist alosetron is available in the US market for the treatment of women with severe, diarrhea-predominant IBS (D-IBS) refractory to conventional therapy, whereas tegaserod, a partial 5-HT4 receptor agonist, has been approved by the FDA and other regulatory agencies for the treatment of women with constipation predominant IBS (C-IBS) or functional constipation. This review is mainly intended to discuss the role of non-neuronal (paracrine) and neuronal 5-HT in the pathophysiology of functional gastrointestinal disorders (FGIDs), such as IBS and functional dyspepsia, and the mechanisms through which drugs acting on 5-HT receptors regulate visceral motility, perception and secretion in these two conditions. PMID- 16699265 TI - CCK1 antagonists: are they ready for clinical use? AB - Cholecystokinin (CCK) is a peptide hormone which is found both in the gastrointestinal tract throughout the human small intestine and nerves in the myenteric plexus of the enteric nervous system and in the central nervous system. This dual location constitutes the anatomical basis for this in functions as a hormone and a neurotransmitter implicated in the regulation of both systems. CCK regulates not only motor functions in the gastrointestinal tract like lower oesophageal sphincter relaxation, gastric secretion and emptying, gall bladder contractility and bile secretion into the duodenum, intestinal and colonic motility, but also sensory functions and plays a role in the regulation of food intake. These effects are mediated through selective receptors CCK1 and CCK2. Over the last few years, research has focused on understanding the role of CCK, its receptors with antagonists at the biological, pharmacological, clinical and therapeutic level. As far as the CCK1 antagonists is concerned, important inroads have been made in the potential role of these antagonists in the treatment of GERD, IBS and pancreatitis. They have also shown encouraging results in sphincter of Oddi dysfunction and some gastrointestinal cancers. This review focuses on the recent ad vances of the biological role of CCK and their CCK1 antagonists: their current basic and clinical status in gastroenterology, with particular emphasis on the potential therapeutic role of the CCK1 antagonists and future research directions. PMID- 16699266 TI - Tachykinin receptors as drug targets for motility disorders. AB - The tachykinins and their receptors are strategically distributed within the gut wall, spinal cord, and central nervous system to be potential targets of therapeutic agents for gastrointestinal motility disorders. However, the development of effective tachykinin receptor agonists or antagonists to treat these disorders has had very limited success so far. This is, in part, due to the complex and multilevel of regulation of gastrointestinal motility function and the challenges faced in targeting the specific type of gut contraction to normalize function in disease state. PMID- 16699267 TI - Peripheral opioids for functional GI disease: a reappraisal. AB - Opioids have been used medicinally and recreationally for thousands of years. The clinical use of opioids for gastrointestinal conditions has been limited by central nervous system side effects. A new generation of peripheral opioid receptor ligands free of central nervous system side effects is being developed. Clinical trials with the peripherally acting mu opioid receptor antagonists' alvimopan and N-methylnaltrexone show promise for improving postoperative ileus- and opioid-induced constipation. Likewise, preliminary studies with the peripherally acting kappa opioid agonist fedotozine showed promise in the treatment of irritable bowel syndrome (IBS) and functional dyspepsia. Further studies are on hold presumably due to lack of efficacy in subsequent studies. However, clinical studies are underway for newer kappa opioid agonists such as asimadoline and ADL 10-0101. PMID- 16699268 TI - Drugs affecting visceral sensitivity: ready for the prime time? AB - Visceral sensitivity has been recognized over the last decade as a frequent pathophysiological component of functional bowel disorders. Studies in animals and humans have identified numerous neurotransmitters involved in the processing of sensations from the gut to the brain. However, up to now none of them has actually been proven to have a marked clinical efficacy and the benefit comes rather from their action of bowel disturbances. Reproducible tests are lacking to detect visceral hypersensitivity in humans and distension tests are difficult to undertake in a clinical setting. Therefore, abnormal visceral sensitivity may not be regarded as a tool to select IBS patients as candidates for a given treatment. PMID- 16699269 TI - Opioid-induced gastrointestinal dysfunction. AB - Use of opioid analgesics is associated with a number of side effects, especially opioid-induced gastrointestinal dysfunction. The extensive use of these compounds and the significant negative impact of the resulting gastrointestinal dysfunction on patients' quality of life make it an important clinical issue. In recent years our understanding of the mechanisms of opioid-induced gastrointestinal dysfunction has advanced greatly. This article reviews the underlying pathophysiological mechanisms of specific gastrointestinal adverse effects of opioids. The role of endogenous opioid peptides in certain gastrointestinal diseases is also discussed. A better understanding of the pathophysiological mechanisms of opioid-induced bowel dysfunction should lead to the development of newer opioid analgesics and improved regimens resulting in reduced gastrointestinal adverse effects. PMID- 16699270 TI - 'Rescue' therapies for the management of Helicobacter pylori infection. AB - Helicobacter pylori infection is the main cause of gastritis, gastroduodenal ulcer and gastric cancer and should be considered as a major public health issue. According to several international guidelines, first-line therapy for treating H. pylori infection consists of proton pump inhibitor (PPI) or ranitidine bismuth citrate (RBC) with any two antibiotics of amoxicillin, clarithromycin or metronidazole given for 7-14 days. However, even with the recommended treatment regimens, approximately 20% of patients will fail to obtain H. pylori eradication. The proportion of patients with first-line H. pylori therapy failure may be higher in clinical practice and it may increase thanks to diffusion of H. pylori treatment. The recommended second-line therapy is the quadruple regimen composed by tetracycline, metronidazole, bismuth salts and a PPI. However, the efficacy of this regimen is limited by poor patient's compliance due to its side effects, number of tablets per day, and long duration. Moreover, bismuth and metronidazole are not available in all countries. Alternatively, a longer-lasting (i.e. 10-14 days) PPI or RBC triple therapy with two antibiotics has generally been used. In an empirical strategy, the choice of second line depends on the treatment initially used. If a clarithromycin-based regimen was administered in first line, a quadruple regimen or PPI (or RBC) triple therapy with metronidazole and amoxicillin (or tetracycline) should be suggested as a second line. In case of second-line treatment failure, the patient should be evaluated by a case-by case approach. A susceptibility-guided strategy, if available, is recommended in order to choose the best third-line treatment. Culture can reveal the presence of H. pylori-sensitive strains to clarithromycin (the best effective) or other antimicrobials (such as amoxicillin, metronidazole and tetracycline). Conversely, in an empirical strategy, a third-line not yet used therapy, can reach a high success rate. PPI or RBC, amoxicillin and a new antimicrobial (e.g. rifabutin, levofloxacin or furazolidone) could be used. Several studies have obtained relatively good results with triple therapy combining PPI, rifabutin, and amoxicillin, although a reversible myelotoxicity as leukopenia and thrombocytopenia has been described. Preliminary good results were also achieved with triples PPI regimens combining levofloxacin and amoxicillin without important adverse effects. Furazolidone has also shown efficacy for H. pylori eradication, although untoward reactions could limit its use, especially when high doses are employed. Finally, in more than one H. pylori treatment failure, non-antimicrobial add-on medications (such as lactoferrin, probiotics and others) could be used with the aim either to improve the eradication rate or to minimize side effects. PMID- 16699271 TI - Emerging biological treatments in inflammatory bowel diseases. AB - Although the advent of infliximab has changed the treatment paradigm and goals in inflammatory bowel disease, it does not provide a cure for it and recent evidence has demonstrated that the immunogenicity of this chimeric anti-tumor necrosis factor antibody is associated with secondary loss of response and intolerance. In ulcerative colitis the efficacy of infliximab was demonstrated in two large clinical trials, but long-term maintenance efficacy data are lacking. Novel biological agents have entered clinical development and pioneering trials have been reported in the last 2 years. For Crohn's disease the fully human IgG1 anti tumor necrosis factor monoclonal adalimumab, and the humanized anti-alpha4 integrin IgG4 antibody, natalizumab have yielded the most promising results in controlled trials, but also agents inhibiting the crucial interleukin 12/interferon-gamma feedback loop suggest therapeutic potential. For severe ulcerative colitis infliximab has been shown to be an effective rescue treatment and the anti-T-cell CD3 antibody has shown promising open-label results. Crucial in the development of novel biological agents, however, is the benefit:risk ratio. As illustrated by unexpected but devastating brain infections with anti adhesion molecules, clinicians should be aware that the powerful immunomodulatory capacity of biologicals necessitates a rigorous safety follow-up. PMID- 16699272 TI - Living drugs for gastrointestinal diseases: the case for probiotics. AB - Nonpathogenic micro-organisms may contain or produce molecules of potential therapeutic interest. This led to the concept of using ingested living micro organisms to produce and transport these molecules to targets in the proximal or distal intestine. Several characteristics of this pharmacological approach are very original: potential for in vivo production of active molecules, for targeting immune cells, for presenting immunogenic molecules in a microbial context, for duodenal delivery using bile sensitivity. Probiotics have been studied for some decades and more recently worm eggs have also received some interest. This paper summarizes facts (especially results of randomized controlled trials and pharmacokinetic studies), and ideas about the use of probiotics to treat or prevent gastrointestinal diseases. The safety of this approach (exceptional cases of infections have been observed), and the potential for using new agents or genetically modified micro-organisms (ongoing trials in humans with Crohn's disease) are also discussed. PMID- 16699273 TI - Pharmacological prevention and treatment of acute pancreatitis: where are we now? AB - Acute pancreatitis is a disease of increasing prevalence, unchanged mortality over many decades, and limited treatment strategies. Progress has been made in developing therapies that reduce the rate of endoscopic retrograde cholangiopancreatography (ERCP)-associated pancreatitis and in preventing infected pancreatic necrosis with intravenous carbapenems. Attempts at reducing pancreatic enzyme output or inhibiting the activity of digestive enzyme proteases have not yielded encouraging results - nor have anti-inflammatory strategies for the treatment of acute pancreatitis been found to be effective so far. Future therapeutic options that are presently being developed or under investigation attempt to restore pancreatic secretory function, interfere with inflammatory pathways in a more effective manner, or inhibit digestive enzyme proteases more selectively. PMID- 16699274 TI - Established and emerging therapies for the treatment of viral hepatitis. AB - Patients who are chronically infected with either hepatitis B or C viruses run the risk of developing cirrhosis and hepatocellular carcinoma in later life. Antiviral treatment offers the only means of interrupting this progression. To date, recombinant interferon alpha and the nucleos(t)ide analogues lamivudine and adefovir dipivoxil are the only licensed drugs for the treatment of chronic hepatitis B, whilst recombinant or pegylated interferons in combination with ribavirin are the ones used for chronic hepatitis C virus infections. The efficacy of these treatments, reasons for treatment failure, drug resistance and future options are discussed in the present review. PMID- 16699275 TI - Emerging therapies for liver fibrosis. AB - Liver fibrosis occurs as a result of a wide range of injurious processes and in its end-stage results in cirrhosis. This gross disruption of liver architecture is associated with impaired hepatic function, portal hypertension and significant resultant morbidity and mortality. Indeed, liver fibrosis and cirrhosis represent a major worldwide healthcare burden. Recent progress in liver transplantation, the management of portal hypertension and the treatment of chronic viral hepatitis have had an important impact. However, these approaches are not without their limitations - in particular, issues regarding organ availability for transplantation - and serve to highlight the urgent requirement to influence pharmacologically the underlying fibrotic process in many patients. Liver fibrosis has been shown to be a bidirectional process and increasing data from laboratory and clinical studies reveal that even advanced fibrosis and cirrhosis are potentially reversible. Exploration of the molecular mechanisms underlying this bi-directionality will lead to char acterisation of the essential attributes of an antifibrotic therapy. In this review, these mechanisms are highlighted and the growing number of emerging antifibrotic agents discussed. PMID- 16699276 TI - Gastroesophageal reflux in healthy subjects induced by two different species of chilli (Capsicum annum). AB - BACKGROUND: Although the ingestion of chilli has been associated with gastroesophageal reflux (GER) symptoms, there are no studies that have explored the effect of a chronic ingestion of different kinds of chilli with a variable content of capsaicin as a cause of GER. METHODS: The effect of chilli on esophageal 24-hour pH monitoring was studied in 12 healthy subjects without GER symptoms before and after of ingestion one of two kinds of chilli. Patients were randomized to ingest 3 g daily of cascabel chilli (Capsicum annum coraciforme containing 880 ppm of capsaicin) or ancho chilli (Capsicum annum grossum containing 488 ppm of capsaicin). RESULTS: After chilli ingestion, the Johnson De Meester Index (JDI) increased significantly [basal: 7 (1-14), after chilli: 13 (2 69), p = 0.0047]. When considering both kinds of chilli separately, the JDI varied, although nonsignificantly, with the ancho chilli [basal: 3 (1-8), after chilli: 10 (2-69), p = 0.11], and significantly with the cascabel chilli [basal: 10 (5-14), after chilli: 18 (2-44), p = 0.028]. CONCLUSION: Our results suggest that the chronic ingestion of chilli induces GER, and that the magnitude of the induced reflux seems to be related to the kind of chilli. PMID- 16699277 TI - NSAID-associated dyspepsia and ulcers: a prospective cohort study in primary care. AB - BACKGROUND AND AIM: Nonsteroidal anti-inflammatory drugs (NSAIDs) cause dyspeptic complaints and lesions in the upper gastrointestinal tract. The true incidence of these side effects in the everyday situation remains uncertain. We therefore investigated as to how often patients on NSAIDs in the primary care setting must be expected to develop troublesome dyspepsia and/or ulcers in the upper gastrointestinal tract. PATIENTS AND METHODS: Admitted to the study were consecutive patients requiring NSAID treatment for at least 2 weeks, who were free of treatment-requiring dyspeptic symptoms, and who were not receiving any prophylactic co-medication. After a minimum of 2 weeks of treatment with a NSAID, a standardized questionnaire and endoscopy of the upper gastrointestinal tract were obtained. RESULTS: 104 patients (median age 53 years, 91 women) were recruited to the study. Four patients had to be excluded for protocol violations. NSAID treatment was applied mainly with diclofenac (n = 67), followed by ibuprofen (n = 22) and rofecoxib (n = 9). The main indication was degenerative complaints affecting the vertebral column and joints. Under treatment, 35% of the patients developed troublesome dyspepsia that required treatment. The frequency of dyspepsia was independent of the duration of NSAID use. Ulcer prevalence was 16% (duodenal ulcer: n = 5; gastric ulcer: n = 11; cardiac ulcer: n = 1). Relevant epigastric pain was experienced more frequently by ulcer patients than those with no ulcer (35 vs. 18%, p = n.s.), but their overall symptom frequency was no higher than in the latter. Predictors for the development of ulcer were smoking (odds ratio 5.11 [1.59-16.48]), regular use of alcohol (odds ratio 4.49 [1.34-15.07]) and duration of treatment less than 1 month (odds ratio 4.95 [1.06 23.09]). No ulcer complications occurred during the period under observation. Overall, 44% of the patients developed troublesome dyspepsia and/or ulcer. CONCLUSION: Primary care patients with an average risk profile frequently develop dyspeptic symptoms requiring treatment, and ulcers while on NSAIDs. Patients who developed an ulcer were not identifiable on the basis of symptoms or risk factors. PMID- 16699278 TI - Rifaximin-based regimens for eradication of Helicobacter pylori: a pilot study. AB - BACKGROUND: Triple therapy is the treatment of choice for Helicobacter pylori infected patients with an eradication rate ranging from 70 to 85%. Poor compliance and antibiotic resistance are the main causes of treatment failure. The aim of the present study was to assess the efficacy of rifaximin, a poorly absorbed antibiotic, for H. pylori eradication. METHODS: We enrolled 48 consecutive H. pylori-positive patients affected. They were randomized to receive two 7-day rifaximin-based triple therapies: rifaximin tablets 400 mg t.i.d., esomeprazole 40 mg o.d. and clarithromycin 500 mg b.i.d. (CRE) or levofloxacin 500 mg o.d. (LRE). H. pylori eradication was assessed using a (13)C-urea breath test 4 weeks after the end of therapy. Treatment compliance and the incidence of side effects were also evaluated. RESULTS: No dropouts were observed. The eradication rate both on intention-to-treat and per-protocol analysis did not show significant differences between groups: 58% (14/24 patients) in group 1 and 42% (10/24 patients) in group 2 (p = 0.24, OR 1.96, 95% CI 0.62-6.18). No significant differences in patients' compliance and incidence of side effects were found between groups. CONCLUSIONS: Rifaximin-based therapy showed optimal compliance but a limited eradication rate compared to standard first-line treatment. Further investigations are needed to evaluate different dosages and combinations. PMID- 16699279 TI - Infliximab therapy improves the bone metabolism in fistulizing Crohn's disease. AB - BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) plays a pivotal role in the pathogenesis of inflammatory bowel diseases and bone resorption as well. Limited data exist about the effect of anti-TNF-alpha infliximab on bone metabolism in inflammatory-type Crohn's disease (CD). AIM: Our aim was to evaluate the effect of infliximab treatment on rapid changes of bone metabolism in fistulizing CD patients. METHODS: 27 patients with fistulizing CD were treated with three series of infliximab. Serum osteocalcin (OC) and beta-CrossLaps (bCL) were measured before administration of each infliximab infusion. 54 patients with inactive CD were controls. RESULTS: In treated patients, there were significant differences in bCL concentrations on days 0 and 14 (p < 0.01) and days 0 and 42 (p < 0.05). OC levels increased significantly between day 0 and 42 (p < 0.05). The values of bCL and OC of control groups differed from serum levels in active patients before the treatment, but not on day 42. Bone markers improved significantly in responder patients, but not in non-responders. CONCLUSION: The beneficial effect of infliximab to the bone metabolism is more expressive in patients whose fistulizing disease improves with this therapy. Our results suggest that TNF alpha has an important role in the alteration of bone metabolism in fistulizing CD patients. PMID- 16699280 TI - Treatment of gastrointestinal stromal tumor with imatinib mesylate: a retrospective single-center experience in Heidelberg. AB - BACKGROUND: Gastrointestinal stromal tumor (GIST) is the most common mesenchymal neoplasm of the gastrointestinal tract. Surgery has been the only effective therapy. However, many patients still eventually die of disease recurrence. Chemotherapy and radiation therapy have been of limited value. Imatinib mesylate (Glivec) is an orally administered competitive inhibitor of tyrosine kinases associated with the KIT, ABL protein, licensed for the treatment of metastatic GIST since 2002 in Germany. METHODS: We summarized the data of 16 patients with advanced or metastatic GIST treated with imatinib mesylate in palliative and neoadjuvant settings. RESULTS: Overall response was 81%, with no evidence of disease (NED) in 3/16 (19%), partial response (PR) in 9/16 (56%) and stable disease (SD) in 1/16 (6%), whereas 3/16 patients (19%) suffered from progressive disease (PD). Mean follow-up was 18.6 months [range: 4-30]. Mean progression-free survival (PFS) was 17.6 months [range: 0-30], mean overall survival (OS) from initial diagnosis was 32.3 months [range: 5-122]. Most common side effects were periorbital edema and skin rash. CONCLUSION: Imatinib mesylate is well tolerated in a dose of up to 800 mg/day and has significant activity during long- term treatment of patients with advanced or metastatic GIST. PMID- 16699281 TI - Association of estrogen receptor alpha (ESR1) PvuII and XbaI polymorphisms with sporadic Alzheimer's disease and their effect on apolipoprotein E concentrations. AB - BACKGROUND: Many studies have shown that estrogen replacement therapy may improve cognitive function in women and reduce the risk of Alzheimer's disease (AD). Because most of the estrogen neuroprotective effect is mediated by receptors, we studied the associations between estrogen receptor alpha (ESR1) polymorphisms (PvuII and XbaI) and AD, and their interactions with apolipoprotein E (APOE) polymorphism and plasma levels. METHODS: ESR1 genotypes and APOE plasma concentrations were determined in a sample of AD patients and controls. RESULTS: ESR1 PP and XX genotypes were associated with an increased risk for AD only in males (OR = 3.6, 95% CI = 1.2-10.9) and conferred a relevant additional risk of AD to subjects also carrying APOE e*4 allele (OR = 13.3, 95% CI = 1.7-103.6). Mean APOE concentrations were lower in AD patients; the lowest levels were observed in male patients carrying PP and/or XX genotypes (p = 0.006) and in patients carrying PP and/or XX genotypes together with the e*4 allele (p = 0.003). In AD women, ESR1 PP and XX genotypes were also associated with lower MMSE values (p = 0.0007). CONCLUSION: The present data suggest that the involvement of ESR1 polymorphisms in AD onset is mediated by the regulation of apoE expression. ESR1 polymorphisms are also associated with a faster cognitive decline in the women AD patients. PMID- 16699282 TI - Apolipoprotein E epsilon4 allele is unrelated to cognitive or functional decline in Alzheimer's disease: retrospective and prospective analysis. AB - OBJECTIVE: The apolipoprotein E (ApoE) epsilon4 allele is a well-documented genetic risk factor for Alzheimer's disease (AD). Its role, if any, in the progression of cognitive and functional impairment in AD has been the subject of discrepant reports in the literature. This study aimed to determine whether ApoE epsilon4 dose is related to the progression of cognitive and functional decline in AD patients by combined retrospective and prospective analyses. METHODS: A sample of 366 AD patients was genotyped for ApoE. Subjects received tests of cognition (Mini-Mental State Examination, MMSE; Alzheimer's Disease Assessment Scale-Cognitive subscale, ADAS-Cog) and daily function (Instrumental Activities of Daily Living, IADL; Alzheimer's Disease Cooperative Study-Activities of Daily Living, ADCS-ADL) at baseline and at multiple subsequent time points during their participation in a variety of research protocols. In retrospective analyses, scores on baseline cognitive and functional measures were compared cross sectionally among genotype groups, controlling for duration of symptoms. In prospective analyses, longitudinal rates of change for each measure were computed by linear regression and compared across genotype groups. RESULTS: No association was observed between ApoE epsilon4 dose and any of the retrospective or prospective measures of cognitive or functional decline in this AD patient sample. CONCLUSIONS: Although ApoE epsilon4 increases the risk for AD and decreases the age of disease onset in population studies, it did not significantly influence the rate of disease progression in cognitive or functional domains in our sample. PMID- 16699283 TI - Retropatellar chondromalacia associated with medial osteoarthritis after meniscus injury. One year of observations in sheep. AB - BACKGROUND/AIMS: In an ovine meniscal repair model, the patellofemoral (PF) osteoarthritis due to a non-sutured tear or failed repair was investigated. METHODS: A radial meniscus tear was either sutured with polydioxanone (PDS), with a slow degrading polylactide long-term suture(LTS) or left without treatment. Knee joint cartilage in the PF and medial compartment was evaluated compared to normal knees (healthy controls). RESULTS: Retropatellar osteoarthritis in the non sutured and sutured animals was intense in contrast to the control knees after 6 months in all groups (p < 0.001), and after 12 months in the PDS group (p < 0.001), LTS group and non-sutured animals (p < 0.05). CONCLUSION: Non-sutured meniscus tears and failed repair lead fast to intense PF osteoarthritis corresponding with tibial damage of the injured compartment. PMID- 16699284 TI - Novices in surgery are the target group of a virtual reality training laboratory. AB - BACKGROUND: This study aims to establish which physicians represent the suitable target group of a virtual training laboratory. METHODS: Novices (48 physicians with fewer than 10 laparoscopic operations) and intermediate trainees (19 physicians who performed 30-50 laparoscopic operations) participated in this study. Each participant performed the basic module 'clip application' at the beginning and after a 1-hour short training course on the LapSim. The course consisted of the tasks coordination, lift and grasp, clip application, cutting with diathermy and fine dissection at increasing difficulty levels. The time taken to complete the tasks, number of errors, and economy of motion parameters (path length and angular path) were analyzed. RESULTS: Following training with the simulator, novices completed the task significantly faster (p = 0.001), demonstrated a greater economy of motion [path length (p = 0.04) and angular path (p = 0.01)]. In contrast, the intermediate trainees showed a reduction of their errors, but without reaching statistical significance. They showed no improvement in economy of motion and completed the task significantly slower (p = 0.03). CONCLUSION: Novices, in comparison to intermediate trainees, tend to benefit most during their first exposure to a laparoscopy simulator. PMID- 16699285 TI - Delayed energy protection of ischemic preconditioning on hepatic ischemia/reperfusion injury in rats. AB - BACKGROUND: Hepatic ischemia/reperfusion (IR) injuries associated with hepatic resections are unresolved problems in the clinical practice. The aim of this study is to elucidate the effect of ischemic preconditioning (IPC) on the energy charge (EC) and related mechanisms at the late phase of hepatic IR injury. METHODS: 30 Wistar rats were randomly divided into sham, IR and IPC groups. The model of partial hepatic IR was used. The rats were subjected to 60 min hepatic ischemia, pretreated by IPC (10/15 min) or not. After 24 h of reperfusion, serum alanine aminotransferase (ALT), nitrite/nitrate (NOx), malondialdehyde (MDA), hepatic tissue arginase activity, adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP) and EC of the liver were measured. RESULTS: Liver injury reduced by IPC is measured by liver tissue arginase activity and serum ALT. Tissue NOx levels in rats pretreated with IPC were significantly higher than levels in the IR group (p < 0.001). Tissue levels of MDA in the liver of the IPC group were found to be significantly lower than the levels in the IR group (p < 0.001). ATP and EC levels 24 h after hepatic ischemia in rats pretreated with IPC were higher than the levels in the IR (p < 0.05). All groups had similar ADP and AMP levels in the liver tissues. The IPC procedure significantly reduced the hepatic necrosis (p < 0.001). CONCLUSION: The results of this study demonstrated that pretreatment with IPC improved tissue ATP, EC, and hepatic necrosis at late stages of ischemia reperfusion injury of the liver. Increased nitric oxide, reduced MDA and arginase activity seemed to play a regulatory role in this delayed protective effect of IPC. PMID- 16699286 TI - Sphincterolysis: a novel approach towards chronic anal fissure. AB - BACKGROUND AND AIMS: The surgical approach in chronic anal fissure is often found associated with disturbed anal continence as well as recurrence. This report describes the author's approach of 'sphincterolysis' or fragmentation of the fibers of the internal sphincter on the left lateral anal wall. PATIENTS AND METHODS: 132 patients with chronic anal fissures were treated with this technique. Pre- and postoperative anal manometry was recorded. The postoperative course and early and 1-year follow-up results were recorded. RESULTS: Early complications included ecchymosis, hematoma, and pain. Fissure healing and relief of symptoms observed in 97% of patients. A transient, variable degree of incontinence occurred in 23 patients and persistent incontinence to flatus and soiling in 5. CONCLUSION: Internal anal sphincterolysis is a safe, effective procedure for the treatment of chronic anal fissure. PMID- 16699287 TI - Allergen-specific polyclonal antibodies reduce allergic disease in a mouse model of allergic asthma. AB - BACKGROUND: Recombinant allergen-specific immunoglobulin G (IgG) antibody therapy can reduce allergic asthma symptoms by inhibiting the immunoglobulin E (IgE) mediated allergic response. This study investigated the effect of intranasally administered allergen-specific monoclonal (mAb) and polyclonal (pAb) antibody on airway inflammation and hyperresponsiveness (AHR) in a mouse model of human asthma. METHODS: Ovalbumin (OVA)-specific IgG2b antibodies were generated by phage display using spleens from OVA-immunized mice, and screening against OVA and finally expressed in CHO cells. Sensitized mice were treated intranasally with either a recombinant anti-OVA mAb (gc32) or a polyclonal preparation comprising seven selected antibodies (including gc32). Control mice received diluent only, OVA only, a control polymeric IgG or dexamethasone. Following challenge with nebulized OVA, investigators assessed airway inflammation by histology and cellular composition of the bronchoalveolar fluid, and methacholine induced airway hyperresponsiveness (AHR). Serum levels of total and OVA-specific IgE were measured by ELISA. RESULTS: Sensitized mice developed airway inflammation and AHR in response to OVA challenge. Intranasally administered OVA specific murine polyclonal or monoclonal IgG2b antibodies both reduced OVA induced lung inflammation. Polyclonal, but not anti-OVA mAb, also reduced AHR and eosinophil influx into the airway lumen. Both anti-OVA antibody preparations reduced levels of specific IgE with no effect on total IgE levels. CONCLUSIONS: Intranasal treatment with allergen-specific pAb reduces pulmonary inflammation and AHR in a mouse model of allergic asthma, but allergen-specific mAb reduces inflammation only. Allergen-specific recombinant pAb offers a potentially valuable therapeutic approach to the management of allergic asthma. PMID- 16699288 TI - Allergic and anaphylactic response to sesame seeds in mice: identification of Ses i 3 and basic subunit of 11s globulins as allergens. AB - BACKGROUND: Allergy to sesame seeds is an emerging food allergy of a serious nature due to a high risk of systemic anaphylaxis. Although a mouse model to study sesame anaphylaxis is desirable, currently it is not available. Here, using a transdermal exposure model system, we tested the hypothesis that sesame seed elicits IL-4-associated IgE antibody response with consequent clinical sensitization in mice. METHODS: Groups of BALB/c mice were exposed to sesame seed extract or saline or a control food (vanilla bean extract) by transdermal applications. Systemic IgE, IgG1 and IgG2a antibody responses were examined using preoptimized ELISA. Type 2 and type 1 cytokine responses were evaluated by ex vivo antigen-mediated activation of spleen cells. Clinical response to oral sesame challenge was studied. Western blot and N-terminal amino acid sequence analyses were performed to identify the sesame allergens. RESULTS: Transdermal exposure to sesame elicited robust IgE and IgG1 but very little IgG2a antibody responses. IgE response to transdermal exposure in two high-IgE responder mice strains with disparate MHC confirmed the intrinsic allergenicity of sesame seed. Transdermal sensitization was associated with activation of IL-4 but not IFN gamma. Furthermore, oral exposure to sesame resulted in clinical signs of systemic anaphylaxis. Western blot and sequence analysis identified four allergens including Ses i 3 and the basic subunit of 11s globulins. CONCLUSION: These data argue that transdermal exposure to sesame seed can result in IL-4 activation, IgE response and clinical sensitization for systemic anaphylaxis. PMID- 16699289 TI - Hyperbaric oxygen treatment augments the efficacy of a losartan regime in an experimental nephrotic syndrome model. AB - BACKGROUND/AIMS: Proteinuria is associated with oxidant stress and inflammation. Hyperbaric oxygen (HBO) treatment has anti-inflammatory and anti-oxidant effects. The aim of the study was to investigate the benefits of HBO treatment on an experimental nephrotic syndrome model. METHODS: 50 male Sprague-Dawley rats weighing 255 +/- 39 g were housed. Forty rats were injected 6 mg/kg adriamycin into tail veins under anesthesia to induce nephrosis, while 10 rats were spared as sham control. After the stabilization of proteinuria at the sixth week, the rats were treated for 6 weeks by losartan (n = 10, 30 mg/kg/day), HBO (n = 10, 2.8 atmosphere absolute, 90 min/day), HBO + losartan (n = 10) and vehicle (n = 10). Protein carbonyl (PCO), superoxide dismutase (SOD) and glutathione peroxidase (GPx) were analyzed from tissue specimens. Biochemical markers were studied from venous samples and 24-hour urine was collected for proteinuria. The surviving animals at 12 weeks (vehicle group (n = 6), HBO (n = 6), losartan (n = 8), HBO + losartan (n = 10) were sacrificed. Glomerular sclerosis, tubulointerstitial and blood vessel changes were determined by semiquantitative scoring. RESULTS: The PCO levels increased (p < 0.001), and the GPx and SOD levels decreased (p < 0.001 for both) in the nephrotic rats. In losartan and HBO groups GPx levels increased (p = 0.001, p = 0.002 respectively), but PCO and SOD levels did not change. The combination of HBO with losartan significantly increased the GPx and SOD levels (p = 0.001 for both) and decreased PCO levels (p = 0.005). HBO but not losartan significantly reduced proteinuria (p < 0.001). The combination of HBO and losartan reduced proteinuria better than the single losartan regime (p < 0.001). The effect of the combination was also noticed on the histological examination of the kidneys. The activities, appetites, weight gains, and improvement of edema were better in the HBO combined with losartan regime. CONCLUSIONS: These results indicate that the addition of HBO therapy to a conventional regime, angiotensin receptor blockers, has significant benefits in the management of proteinuria. Future clinical studies are needed to elucidate the role of HBO and other antioxidant strategies in the treatment of proteinuria. PMID- 16699290 TI - Estradiol, but not dehydroepiandrosterone, decreases parasitemia and increases the incidence of cerebral malaria and the mortality in plasmodium berghei ANKA infected CBA mice. AB - OBJECTIVE: The effect of castration and subsequent replacement of dehydroepiandrosterone (DHEA) or estradiol on parasitemia, mortality and incidence of cerebral malaria (CM) was evaluated in CBA mice infected with Plasmodium berghei ANKA. METHODS: Female mice were castrated, and groups of 12-15 animals received daily injections of DHEA, estradiol or saline. Four days after the start of treatment, mice were inoculated with 1 x 10(6)P. berghei ANKA parasitized erythrocytes. DHEA treatment was continued during the 5 days after infection, and estradiol was administered during the follow-up. Parasitemia was evaluated daily in Giemsa-stained blood smears. Signs of CM were determined by the manifestation of coma, limb paralysis and/or convulsions. Plasma TNF-alpha levels were evaluated by sandwich ELISA. Nitric oxide synthase (NOS) activity in the brain of moribund mice was measured by the method of Bredt and Snyder. RESULTS: In non-castrated infected mice, the incidence of CM was 50%, and plasma TNF-alpha increased and brain NOS activity decreased compared to non-infected controls. Castration had no major effect on the parameters analyzed (parasitemia, mortality, CM incidence, TNF-alpha levels or NOS activity). Estradiol replacement caused a decrease in parasitemia but resulted in higher CM incidence and faster mortality, with an increase in NOS activity. CONCLUSIONS: Estradiol modulated the immune response of P. berghei ANKA-infected CBA mice, decreasing parasitemia and increasing NOS activity, and impacted negatively on survival and CM incidence, showing that neuroimmunoendocrine interactions are important in the physiopathogenesis of malaria infections. PMID- 16699291 TI - Antidiuretic hormone and osmolality in isosorbide therapy and glycerol test. AB - Changes in plasma antidiuretic hormone (p-ADH) and plasma osmolality (p-Osm) levels were studied in 63 patients with Meniere's disease before and after isosorbide administration and a glycerol test. Increments in both p-ADH and p-Osm levels were observed after isosorbide administration and the glycerol test. The p ADH secretion appeared to be secondarily stimulated by an increase in the p-Osm level. This p-ADH level increase affects cochlear fluid homeostasis. Based on these results, we must consider both the p-Osm and p-ADH levels when treating patients with Meniere's disease by osmotic diuretics. PMID- 16699292 TI - Effect of oral administration of crude aqueous extract of garlic on pharmacokinetic parameters of isoniazid and rifampicin in rabbits. AB - AIM: To study the effect of oral administration of crude aqueous extract of garlic for 14 days on pharmacokinetic parameters of isoniazid and rifampicin. MATERIALS AND METHODS: Crude extract was prepared according to the method described by Fromtling and Bulmer. The study was done on 16 New Zealand white rabbits, divided into two groups of 8 animals each for two drugs. Baseline pharmacokinetic parameters for single-dose isoniazid and rifampicin were calculated from plasma drug concentrations obtained at various time intervals after dosing. The animals were given garlic extract orally for 14 days. Pharmacokinetic parameters were calculated again as done previously. OBSERVATIONS: Administration of crude aqueous extract of garlic significantly altered the pharmacokinetic parameters for isoniazid. C(max) was reduced from 15.4 +/- 5.6 to 5.4 +/- 3.3 microg/ml. AUC((0-24)) was reduced from 76.7 +/- 25.0 to 34.3 +/- 19.2 microg/ml.h. No significant change in T(max), k(el) and AUC((0 )(alpha)) was seen. Pharmacokinetic parameters of rifampicin were not significantly altered by administration of garlic extract. CONCLUSIONS: Oral administration of garlic extract decreased the bioavailability of isoniazid significantly with no change in rate of elimination. Bioavailability of rifampicin is not significantly altered by garlic extract. PMID- 16699293 TI - Six cases of ectopic ACTH syndrome caused by thymic carcinoid. AB - The ectopic ACTH syndrome (EAS) caused by thymic carcinoid is rare and its diagnosis remains a challenge to the endocrinologist. Here are six cases of EAS with a typical Cushing habitus accompanied by hyperpigmentation and hypokalemia. For all six patients, the high dose (8 mg) dexamethasone suppression test (HDDST) showed lack of suppression, computed tomography (CT) scanning documented anterior mediastinal masses, and the mediastenal tumors removed were confirmed as ACTH secreting thymic carcinoids by positive ACTH and NSE staining. Our data indicate that HDDST chest radiologic imaging and other laboratory examinations will greatly assist in diagnosing the thymic carcinoid-induced EAS at an earlier stage, which will significantly improve the long-term survival of the patient. PMID- 16699294 TI - Prevalence of adrenal incidentaloma in a contemporary computerized tomography series. AB - Adrenal incidentalomas, defined as masses discovered incidentally during imaging investigation of non-adrenal disorders, have become a rather common finding in clinical practice. The prevalence is not well characterized and varies among studies. The aim of the present study was to perform a prospective evaluation of the prevalence of adrenal incidentalomas among subjects undergoing computerized tomography (CT) scan of the chest in a screening program of lung cancer (Tic TAC study) in Piedmont, a region of Northwestern Italy. This evaluation included 520 subjects (382 males and 138 females, aged between 55-82 yr), referred to our hospital from April to December 2001. Twenty-three patients with adrenal masses were identified: 21 adrenal adenomas, 1 myelolipoma, and 1 metastasis of lung cancer. Therefore, the overall prevalence of adrenal lesions was 4.4%, and that of benign adrenal masses was 4.2%. This prevalence is higher than those found in previous CT scan series reported in the literature, probably because of the use of high-resolution CT scanning technology. Another factor that influenced our results is that subject age is skewed towards the decades characterized by a greater occurrence of adrenal masses. The outcome of this study confirms that we are presently able to identify incidentally discovered adrenal masses more often than in early years and that the prevalence of adrenal incidentalomas on CT images is approaching that of autopsy series. The present study provides a reliable estimate of the prevalence of adrenal incidentaloma with currently used CT scanners. Notwithstanding that our subjects were at increased risk of lung cancer, the rate of adrenal metastases was low. We think that the present results can be generalized even if we may disclose the lack of histological diagnosis. PMID- 16699295 TI - Changes in calcium, 25(OH) vitamin D3 and other biochemical factors during pregnancy. AB - INTRODUCTION AND AIMS: Calcium and vitamin D play major roles in calcium homeostasis and skeletal development, especially during pregnancy. This study was conducted to determine changes in calcium, 25 hydroxy [25(OH)] vitamin D3 and other biochemical factors (PTH, osteocalcin, alkaline phosphatase, magnesium, phosphorus) related to calcium homeostasis and bone turnover during pregnancy and compare the values to those of non-pregnant women. MATERIALS AND METHODS: In a cohort study, 48 pregnant women, in their first trimester of pregnancy (12+/-2.7 weeks), from 5 prenatal care centers, and 47 non-pregnant women randomly selected from the Tehran Lipid and Glucose Study (TLGS) population were enrolled. These pregnant women were followed in their second (26+/-1.9 weeks) and third trimesters (37+/-3.2 weeks) of pregnancy. Samples were drawn from June 2002 to March 2003. Including criteria were healthy women with no background of disease. Women using photo protection and calcium and vitamin D supplementation were excluded. A questionnaire was used to obtain demographic information for both groups. Venous blood samples were taken after 12-14 h of overnight fasting to measure serum calcium, phosphorus, magnesium, alkaline phosphatase, PTH, 25 (OH) vitamin D3 and serum osteocalcin levels. The repeated measures analysis of variance and t-test were used for statistical analysis. Data were matched for age and weight in both the case (in the first trimester) and control groups. RESULTS: Significant differences were found in the mean serum levels of osteocalcin and alkaline phosphatase between the three trimesters of pregnancy (p< 0.001). Osteocalcin was significantly higher in the first trimester as compared to second and third trimesters of pregnancy. Alkaline phosphatase was significantly lower in the first trimester as compared to the second and third trimesters of pregnancy and their controls. There was also a significant difference in osteocalcin in the second and third trimesters and alkaline phosphatase in the first and third trimesters of pregnancy in comparison to the control group. The mean values of osteocalcin were 12.7+/-8.5, 8.1+/-6.9, 5.6+/-5.0 and 13.9+/-7.9 ng/ml, respectively, and mean values for alkaline phosphatase were 115+/-38, 125+/-37, 174+/-61 and 134+/-35.0 Iu/l, respectively. In the first trimester, alkaline phosphatase was lower and osteocalcin was higher than in the second and third trimesters. In the first trimester of pregnancy, 20 and 40% of women had 25(OH) vitamin D3 < 10 and < 20 ng/ml, respectively, and 19% of women had serum calcium levels < 8.6 mg/dl. CONCLUSION: 60% of women in the first trimester, 48% in the second and 47% in the third trimester had either severe or moderate vitamin D deficiency. It is recommended that the importance of calcium supplements with vitamin D in pregnant women be stressed for these individuals. PMID- 16699296 TI - Serum PSA levels are not affected by the menstrual cycle or the menopause, but are increased in subjects with polycystic ovary syndrome. AB - BACKGROUND/AIMS: Prostatic specific antigen (PSA) is the most specific prostatic tumor marker in man. Recently, PSA has been detected in a variety of tissues and fluids in women, and its determination suggested as a marker of hyperandrogenism. However, precise information about the physiology of PSA in females is not available. The goal of this study was to assess serum concentrations of PSA in healthy pre-menopausal women (healthy pre-menopausal group), menopausal women (menopause group) and patients with polycystic ovary syndrome (PCOS group). METHODS: PSA, androgens, LH, FSH, 17-beta-estradiol (E2), progesterone (Pg) were assessed in 40 post-menopausal women, 35 fertile controls and 35 women with PCOS. RESULTS: No significant difference in PSA concentrations could be demonstrated in different phases of the menstrual cycle in healthy pre-menopausal group and between pre- and post-menopausal groups. No correlations could be demonstrated between serum PSA levels and the following parameters: age, body mass index (BMI), LH, FSH, E2, testosterone (T), DHEAS, and SHBG, both in pre- and post menopausal women. Significantly higher PSA levels (median=14 pg/ml) were found in the PCOS group compared to both pre-menopausal (median=5 pg/ml) and menopausal (median= 5 pg/ml) groups (p< 0.05). CONCLUSIONS: only minor fluctuations of serum PSA concentrations are observed in healthy pre- and post-menopausal women, while serum level is higher in PCOS, and therefore PSA can be considered a suitable marker of female hyperandrogenism. PMID- 16699297 TI - A rural-urban comparison of the characteristics of the metabolic syndrome by gender in Korea: the Korean Health and Genome Study (KHGS). AB - The number of cases of the metabolic syndrome is increasing dramatically in Western countries. However, the evaluation of the metabolic syndrome is limited in Asian countries. Thus, our objectives were: 1) to investigate parameters of the metabolic syndrome defined by the National Cholesterol Education Program (NCEP)-Adult Treatment Panel III (ATPIII) in the subjects representing Korean general population and 2) the modification of which factor is most effective in reducing the metabolic syndrome. A total of 10,044 (5024 rural and 5020 urban) Korean men and women in the age range 40-69 yr voluntarily participated in this community-based cross-sectional study (a rural and an urban community was selected). Anthropometric parameters (weight, height, waist and hip circumference and blood pressure), social factors (smoking, alcohol, exercise and education status) as well as biochemical parameters (fasting glucose and insulin, lipids and body composition) were measured. Twenty-six point one per cent of the total subjects were classified as having the metabolic syndrome. Age- and sex-adjusted prevalences were 29.3 and 22.3% in the rural and urban community, respectively (p< 0.01). Abdominal obesity (46.9%) and high blood pressure (45.2%) were major components in the rural community; hypertriglyceridemia (37.6%) and low HDL cholesterolemia (37.0%) in the urban community. In conclusion, abdominal obesity in the rural community and dyslipidemia in the urban community should be a main subject of intervention, aimed at reducing the prevalence of the metabolic syndrome in Korea. Given the rapid progression of the Korean economy over the past 30 yr, the prevalence of the metabolic syndrome is expected to increase continuously. A strategy to prevent this expected extraordinary event should be conducted at a national level. PMID- 16699298 TI - Erythropoietin in thyroid cancer. AB - Erythropoietin (Epo) and the epo-receptor (EpoR) have been implicated in tumor growth, invasion and metastasis. We previously demonstrated Epo and EpoR expression in a small group of archived papillary thyroid cancers (PTC), but were unable to examine functional integrity using formalin-fixed tissues. In the present study, we examined the in vitro expression, induction and function of Epo and EpoR in papillary (NPA), follicular (WRO) and anaplastic (ARO-81) thyroid cancer cells. We found that all three cell lines expressed Epo and EpoR mRNA and that the hypoxia-mimetic cobalt induced Epo expression in all cell lines. None of the growth factors we examined (thyrotropin, vascular endothelial growth factor, IGF-I, or human Epo) altered Epo or EpoR gene expression. Importantly, however, administration of Epo to NPA but not WRO cells resulted in significant alterations in the expression of several mitogenic genes including cyclooxygenase 2 (COX-2), beta-casein (CSN2), wild type p53-induced gene-1 (WIG1) and cathepsin D (CTSD). Epo treated ARO-81 cells only had an increase in CSN2 expression. We conclude that Epo and EpoR are expressed by thyroid cancers and that stimulation of the Epo/EpoR signal pathway results in changes that could impact on the clinical behavior of thyroid cancers. PMID- 16699299 TI - A study on glucose metabolism in a small cohort of children and adolescents with kidney transplant. AB - Post-transplant diabetes mellitus (PTDM) and impaired glucose tolerance are now considered among the major adverse events following organ transplantation. The present study was aimed at investigating the regulation of glucose metabolism in pediatric recipients of a kidney transplant (KT), receiving tacrolimus or cyclosporine A-based immunosuppression. Twelve subjects, eight males and four females, aged 12.1+/-3.8 yr, and with a mean time from KT of 45.6 months were enrolled in the study. All patients had a basal evaluation of fasting glucose (GF), fasting insulin (IF), C-peptide and glycated hemoglobin (HbA1c) levels. They then underwent oral glucose tolerance test (OGTT), with measurement of blood glucose and insulin concentration. Two children had impaired GF, associated with supernormal HbA1c levels, one patient showed impaired glucose tolerance, none had PTDM. Peripheral insulin resistance, as measured by quantitative insulin sensitivity check index (QUICKI) and homeostasis model assessment estimate of insulin sensitivity (HOMA-IR) index, was enhanced in 3 patients. Subsequently, GF significantly increased with time from transplant (p=0.01), while fasting C peptide and the area under the curve of insulin correlated with creatinine clearance. In conclusion, our results, although generated in a small sample size, would suggest that long-term follow-up of children receiving a KT should extend to explore the response to oral glucose load and at least the basal measure of insulin response. PMID- 16699300 TI - Human placenta and fetal membranes express nerve growth factor mRNA and protein. AB - The present study investigated whether trophoblast, decidua and fetal membranes express nerve growth factor (NGF) mRNA and peptide. Tissue specimens were collected in the first and third trimester of pregnancy from women undergoing voluntary pregnancy interruption (no.= 6; from 8 to 12 gestational weeks) and from women having an elective caesarean section at term (no.= 6; week 39-40 of pregnancy). Using reverse transcriptase-polymerase chain reaction (RT-PCR), trophoblast, amnion/chorion and maternal decidua showed the expression of NGF mRNA both in early gestation and at term. By immunohistochemistry, the immunoreactive NGF was found in the cyto and syncytial trophoblast cells, chorionic mesodermic cells and in decidua. Vessel endothelial cells were stained in maternal compartments, while fetal vessels were unstained. These results, showing the expression and localization of NGF, support the current concept that human placenta is a potent neuroendocrine organ throughout gestation. PMID- 16699301 TI - One-tube-PCR technique for CCL2, CCL3, CCL4 and CCL5 applied to fine needle aspiration biopsies shows different profiles in autoimmune and non-autoimmune thyroid disorders. AB - Autoimmune thyroid diseases are characterized by lymphocytic infiltration of the thyroid gland. Chemokines are crucial in the recruitment of lymphocytes and might play an important role in the pathogenesis of autoimmune thyroid disease. The aim of this study was to test the feasibility of analysing by one-tube reverse transcriptase polymerase chain reaction (RT-PCR) technique CC chemokine profiles in samples obtained by fine needle aspiration biopsy (FNAB). In 27 out of 35 (77%) samples, the material was sufficient for analysis and in 16 (59%) chemokines were detected, thus demonstrating the potential of this technique. Moreover, even in this small group, a statistically significant increase of CCL3 and CCL4 was found in samples from patients with autoimmune thyroid disease as compared to those with multinodular goiter. Chemokine profile measured by improved multiamplification techniques in FNAB thyroid samples may become a useful complementary tool for the management of thyroid autoimmune disease as it constitutes a source of data for research of their pathogenesis. PMID- 16699302 TI - The malignant potential of a succinate dehydrogenase subunit B germline mutation. AB - Familial catecholamine secreting tumors have been associated with multiple endocrine neoplasia type 2, Von Hippel-Lindau disease and neurofibromatosis type 1. In the last years, mutations of genes encoding subunits B, C and D of the succinate dehydrogenase have been discovered as other causes of pheochromocytomas and paragangliomas. We diagnosed a malignant retroperitoneal paraganglioma in a 64-yr-old man with bone metastasis in 2001. Two years later a retroperitoneal benign paraganglioma was found and resected in his 32-yr-old daughter. Thus we diagnosed in this family a paraganglioma syndrome. We performed molecular genetic analyses of the genes SDHB, SDHC, and SDHD. We detected in the SDHB gene the mutation SDHB c. 558-3 C> G affecting the splice site of exon 5. In a second daughter the mutation was also detected, thorough clinical investigation revealed normal results. We conclude that the SDHB mutation predisposes to abdominal extra adrenal and potential malignant pheochromocytoma with incomplete penetrance. PMID- 16699303 TI - Hypophysitis associated with a ruptured Rathke's cleft cyst in a woman, during pregnancy. AB - We report the case of a 29-yr-old woman who first presented an aseptic meningitis at the beginning of a pregnancy. She was admitted one month later with headaches and vomiting. Panhypopituitarism with diabetes insipidus was diagnosed. Magnetic resonance imaging (MRI) data suggested the existence of lymphocytic infundibulohypophysitis, with inflammation of the suprasellar area. No new symptoms were noticed until 6 months later when this patient pointed out troubles of the visual field, due to a compression of the optic chiasma. Three boluses of 1 g methylprednisolone were prescribed, with no effects. After delivery, the defects of the visual field increased. A neurosurgical intervention was decided. Diagnosis of Rathke's cleft cyst (RCC) was made. We concluded that this patient presented a rupture of a RCC, which occurred at the beginning of pregnancy, associated later with panhypopituitarism with diabetes insipidus, due to a probable hypophysitis. The end of the pregnancy was marked by consequences of an increased volume of the RCC. To our knowledge, this case is the first described during pregnancy. PMID- 16699304 TI - Long-term quiescence of ectopic Cushing's syndrome caused by pulmonary neuroendocrine tumor (typical carcinoid) and tumorlets: spontaneous remission or therapeutic effect of bromocriptine? AB - In 1990, a 55-yr-old woman was admitted to the Medical Department of our hospital for severe hypercortisolism complicated by secondary diabetes mellitus and serious hypokalemia. Although inferior petrosal sinus sampling did not show any significant difference between central and peripheral ACTH concentration, suggesting an ectopic source of ACTH secretion, diagnostic imaging was negative and Cushing's disease due to hyperplasia of the pituitary intermediate lobe was suspected. Medical treatment with bromocriptine and cyproheptadine led to a rapid and stabile normalization of adrenal function, so that after two months cyproheptadine was stopped and bromocriptine was tapered to a smaller dose. An attempt to discontinue medical treatment, carried out 3 yr later, was followed by a quick increase of ACTH and cortisol levels, which were normalized by the resumption of the bromocriptine. Adrenal function remained normal until 1994 when hypercortisolism relapsed despite the treatment. Chest radiography and computed tomography (CT) scan detected a 6 mm nodule in the middle lobe of the lung which proved to be a neuroendocrine tumor, with immunohistochemical positivity for ACTH. Nests of neuroendocrine cells (tumorlets) were also demonstrated in the surrounding lung tissue. After the lobectomy, the patient recovered completely from Cushing's syndrome and no symptoms and/or signs of recurrence have been observed over the subsequent follow-up period. Although cyclical spontaneous Cushing's syndrome could not be excluded, there was strong evidence that medical treatment with bromocriptine might have played a key role in long-lasting remission. To our knowledge, this is the second case described in literature of Cushing's syndrome caused by neuroendocrine lung tumor responsive to bromocriptine. PMID- 16699305 TI - Severe reversible dilated cardiomyopathy in a patient with multiple endocrine neoplasia 2A syndrome. AB - Pheochromocytoma may infrequently lead to dilated cardiomyopathy, which may reverse partially or completely after treatment. Progressive dyspnea, palpitations, and paroxysmal attacks of severe hypertension leading to cardiac failure had developed in a 25-yr-old woman. Chest radiography and echocardiography revealed a massive 4-chamber dilatation of the heart with an ejection fraction of 12%. Twenty-four-h urinary vanillylmandelic acid and metanephrine levels were elevated. Magnetic resonance imaging detected a large mass lesion in the right adrenal gland. Oral glucose tolerance testing revealed diabetes mellitus. Medical drug therapy with alpha-blocker, angiotensin converting enzyme inhibitor, beta-blocker, digoxin, and diuretic rapidly improved her cardiac condition. Repeat echocardiogram showed that the left ventricular function had improved substantially. The clinical condition of excess catecholaminemia (and thus, arterial hypertension and the abnormality of the glucose metabolism) subsided with complete resolution of the congestive heart failure following the surgical removal of the tumor. Evaluation for medullary thyroid carcinoma (MTC) revealed an elevated calcitonin level demonstrated by fine needle aspiration biopsy. There were no biochemical evidences for primary hyperparathyroidism. Multiple endocrine neoplasia 2 (MEN 2A) syndrome was diagnosed. An overwhelming secretion of catecholamine might cause severe cardiomyopathy and impair glucose metabolism, as evidenced by the improvement of both conditions following the medical treatment of catecholaminemia and surgical resection of the tumor. PMID- 16699306 TI - A case of reversed pituitary dysfunction with intrasellar mass. AB - Hypopituitarism can be caused by tumor, inflammation, granuloma and injuries. Once pituitary function is disturbed, hormone replacement therapy is necessary for the remaining life span in most cases. We have experienced a rare case of a unique intrasellar mass associated with pituitary dysfunction in which both spontaneously reversed. A 61-yr-old woman developed hypoadrenalism and central diabetes insipidus (cDI). Magnetic resonance (MR) imaging revealed a lobular, strong hypointense lesion with spotty signal in the middle of the hypophysis. This spotty lesion showed isointensity on T1- and high-intensity on T2-weighted MR images. The spotty signal as well as the normal pituitary lobe were enhanced by the administration of gadolinium. As replacement therapies for hypoadrenalism and cDI, 10 mg of hydrocortisone and 2.5 microg of desmopressin acetate were prescribed. Three months later, slight shrinkage of intrasellar mass and spontaneous improvement of pituitary functions were found. Hydrocortisone was then discontinued. Furthermore, because polyuria and polydipsia were improved nine months later, desmopressin acetate was stopped. Currently, the intrasellar mass continues to shrink, and the patient shows no symptoms without medication. Based upon the unique features of MR images, we suspect that the origin of the mass is an intrasellar hemangioma. PMID- 16699307 TI - Aldosterone and the conquest of land. AB - The sequence of the phylogenetic events that preceded the appearance of aldosterone in vertebrates is described, starting from the ancestral conversion of cytochrome P450s from oxygen detoxification to xenobiotic detoxification and synthesis of oxygenated endobiotics with useful functions in intercellular signalling, such as steroid hormones. At the end of the Silurian period [438-408 million yr ago, (Mya)], a complete set of cytochrome P450s for corticoid synthesis was presumably already available, except for mitochondrial cytochrome P450c18 or aldosterone synthase encoded by CYP11B2. This gene arose by duplication of the CYP11B gene in the sarcopterygian or lobe-finned fish/tetrapod line after its divergence from the actinopterygian or ray-finned fish line 420 Mya, but before the beginning of the colonization of land by tetrapods in the late Devonian period, around 370 Mya. The fact that aldosterone is already present in Dipnoi, which occupy an evolutionary transition between water- and air breathing but are fully aquatic, suggests that the role of this steroid was to potentiate the corticoid response to hypoxia, rather than to prevent dehydration out of the water. In terrestrial amphibians, there is no differentiation between the secretion rates and gluco- and mineralocorticoid effects of aldosterone and corticosterone. In sauropsids, plasma aldosterone concentrations are much lower than in amphibians, but regulation of salt/water balance is dependent upon both aldosterone and corticosterone, though sometimes with opposed actions. In terrestrial mammals, aldosterone acquires a specific mineralocorticoid function, because its interaction with the mineralocorticoid receptor is protected by the coexpression of the enzyme 11beta-hydroxysteroid dehydrogenase type 2, which inactivates both cortisol and corticosterone. There is evidence that aldosterone can be also synthesized extra-adrenally in brain neurons and cardiac myocytes, which lack this protection and where the effects of aldosterone oppose those of glucocorticoids. In conclusion, the phylogenetic history of aldosterone documents the erratic progression of evolutionary changes in the course of the strenuous struggle for environmental resources and survival. PMID- 16699308 TI - Restriction of dietary Iodine does not ameliorate the early effect of anti thyroid drug therapy for Graves' disease in an area of excessive iodine intake. AB - The close relationship between iodine intake and the effects of anti-thyroid drugs (ATD) for Graves' disease (GD) has been well established. However, it remains unknown whether restriction of dietary iodine improves the effect of ATD. This study aimed to clarify this issue in Japanese patients with GD who routinely ingest large amounts of dietary iodine. We performed a prospective clinical study in 81 patients with newly diagnosed GD who were divided into an iodine restricted group and a control group. Urinary iodine, thyroid hormones and TSH receptor antibody were measured during the first 8 weeks of ATD therapy. Urinary iodine concentrations in the iodine restricted group were significantly lower than in the control group (p=0.043). However, there were no significant differences in the decrease of thyroid hormones and TSH receptor antibody between the two groups. Restriction of dietary iodine does not ameliorate the effect of ATD therapy for GD in an area of excessive iodine intake. PMID- 16699309 TI - Pseudomyxoma peritonei of appendiceal origin: a clinicopathologic analysis of 101 patients uniformly treated at a single institution, with literature review. AB - Pseudomyxoma peritonei is a clinical term for gelatinous ascites, usually secondary to an appendiceal tumor. The pathologic classification of pseudomyxoma peritonei and its associated appendiceal tumors has been plagued with controversy and confusing terminology. In an effort to clarify this, we reviewed the pathology of 101 patients, all treated at our institution from 1993 to 2005, with pseudomyxoma peritonei of appendiceal origin. All patients were uniformly treated with our standardized protocol. This is the largest pathologic series solely devoted to appendiceal neoplasia with gelatinous ascites. The cases were assigned, according to previously published criteria, to the categories of disseminated peritoneal adenomucinosis (DPAM), peritoneal mucinous carcinomatosis (PMCA), or PMCA with intermediate (well differentiated) features (PMCA-I), with the exception that any case with a signet-ring cell component was considered as PMCA and not PMCA-I. By histologic category, 58 patients had DPAM, 23 were PMCA, and 20 were PMCA-I.One-year, 3-year, and 5-year survival outcomes were not significantly different between DPAM and PMCA-I. DPAM and PMCA-I also exhibited a roughly equal incidence of parenchymal (beyond the serosa) organ invasion. Survival outcomes were significantly worse for PMCA, compared with PMCA-I and DPAM. After reviewing our data and the literature, mucinous carcinoma peritonei low grade was applied to the low-grade histology of pseudomyxoma peritonei, including those cases referred to by some as DPAM in the same category as PMCA-I. Cases that are moderately differentiated to poorly differentiated are classified as mucinous carcinoma peritonei-high grade. PMID- 16699310 TI - Microadenomatosis of the endocrine pancreas in patients with and without the multiple endocrine neoplasia type 1 syndrome. AB - It has been suggested that microadenomatosis of the endocrine pancreas is a hallmark of the multiple endocrine neoplasia type 1 syndrome (MEN1). This study attempts to elucidate the relationship between pancreatic microadenomatosis and the MEN1 and von Hippel-Lindau (VHL) syndromes. Pancreatic tissue specimens from 37 patients (with either microadenomatosis or the MEN1 syndrome) were analyzed using immunohistochemistry, confocal laser scanning microscopy, and morphometric methods. The MEN1 and the VHL status were assessed on the basis of clinical criteria (all patients) and PCR-based mutational analysis (15 and 5 patients, respectively). Pancreatic microadenomatosis was found in 35 of 37 patients, 28 of whom fulfilled the clinicopathologic criteria and 13 the genetic criteria for MEN1, whereas none of the patients had evidence of a VHL syndrome. Microadenomas were present in 26 of the 28 MEN1 patients, and all these tumors were consistently multihormonal. Five of the 9 patients with microadenomatosis and no clinical evidence for MEN1 or VHL also lacked mutations for the respective genes. Five of these 9 patients suffered from hyperinsulinism and revealed multiple insulin-positive tumors. The other patients were nonsymptomatic and showed multiple glucagon-expressing neoplasms. In microadenomatosis patients with and without the MEN1 syndrome, a subset of morphologically normal-appearing islets showed increased endocrine cell proliferation. In conclusion, endocrine multihormonal microadenomatosis of the pancreas is a feature of MEN1. In addition, a monohormonal type of pancreatic microadenomatosis was identified that consisted of either insulinomas or glucagon-producing tumors and was not associated with MEN1 or VHL. PMID- 16699311 TI - Evaluation of the nonneoplastic pathology in tumor nephrectomy specimens: predicting the risk of progressive renal failure. AB - Pathologic changes in nonneoplastic renal parenchyma of nephrectomy specimens for renal tumors and the significance of these changes with regard to the outcome of contralateral kidney function have not been studied previously. We examined the nonneoplastic renal parenchymal changes in 110 consecutive tumor nephrectomy specimens, and we correlated our findings with patients' clinical information. The material was examined for the presence of any glomerular, tubulointerstitial, or vascular pathology. In our analysis, only about 10% of cases had unremarkable renal parenchyma and vasculature. A further 28% of cases had unremarkable parenchyma, but some degree of vascular sclerosis was noted. The remaining cases (>60%) had evident pathologic abnormalities, most commonly related to vascular disease or diabetes mellitus. Regardless of the type of renal cancer they have, the majority of our cases can be placed in one of three principal groups: 1) unremarkable kidney parenchyma, with or without vascular sclerosis (38%); 2) parenchymal scarring and marked vascular changes, including cases of atheroembolic disease, and chronic thrombotic microangiopathy (28%); and 3) changes related to diabetes mellitus, such as glomerular hypertrophy, mesangial expansion, and diffuse glomerulosclerosis (24%). Follow-up data on serum creatinine 6 months postoperatively were available in a third of our patients. Patients with severe histopathologic findings (parenchymal scarring with >20% global glomerulosclerosis and advanced diffuse diabetic glomerulosclerosis) showed a significant change in serum creatinine from the preoperative period to 6 months after radical nephrectomy (P=0.001), indicative of progressive worsening of renal function; this change is significantly greater than that seen in patients with unremarkable renal parenchyma (P=0.01). We conclude that adequate examination of nonneoplastic renal parenchyma is an important tool in recognizing patients at risk for progressive renal disease after nephrectomy and could be an essential step in providing early preventive and treatment measures and better medical care of patients undergoing nephrectomy for neoplastic processes. PMID- 16699312 TI - IgD positive L&H cells identify a unique subset of nodular lymphocyte predominant Hodgkin lymphoma. AB - Nodular lymphocyte predominant Hodgkin lymphoma (NLPHL) is a rare B-cell lymphoma considered to be of germinal center (GC) derivation. Studies on immunoglobulin expression have been few, and post-switch immunoglobulin (IgG) has been identified in the majority of cases examined thus far. We reviewed 180 cases of NLPHL and observed the unexpected expression of IgD in 27% of cases. IgD is usually coexpressed with IgM in naive B cells but can also be seen as IgD-only in centroblasts (CD38-positive) or memory B cells (CD27-positive). We asked whether IgD-positive NLPHL differed from cases of NLPHL negative for IgD. Clinically, the IgD-positive cases presented at a younger median age (21 vs. 44 years) and had a striking male predominance (male-to-female ratio, 23:1 vs. 1.5:1). Cervical lymph nodes were more frequently involved (56% vs. 18.2%). L&H cells were localized in a predominantly extrafollicular distribution in the majority of IgD-positive cases (69%). The IgD-positive cases did not coexpress IgM or CD27 (a marker associated with memory B cells), and nearly all (93%) were weakly positive for CD38, supporting a GC derivation. The expression of Bcl-6, BOB.1, Oct2, and SWAP 70 was similar in the two groups. However, PU.1 expression was seen in 60% of the IgD-positive cases in contrast to 86% of the IgD-negative cases. The absence of PU.1 staining correlated with more L&H cells in an extrafollicular distribution, weakening the use of this marker in the differential diagnosis with T-cell rich/histiocyte rich B-cell lymphomas. To study IgD expression in "de-novo" T cell rich/histiocyte rich B-cell lymphomas, we analyzed 20 cases and all but one were negative. In conclusion, cases of IgD-positive NLPHL do not differ from IgD negative cases regarding cellular derivation and most other immunophenotypic characteristics. However, IgD-positive NLPHL exhibits distinctive clinical features, and more often involves the interfollicular region in a background relatively rich in T cells. IgD positivity may represent an additional useful marker in the diagnosis of NLPHL. PMID- 16699313 TI - Angiomyolipoma with epithelial cysts (AMLEC): a distinct cystic variant of angiomyolipoma. AB - Renal angiomyolipoma (AML) is typically a solid lesion, composed of varying amounts of adipose, vascular, and muscular tissue, lacking an epithelial component. Although it is known that entrapped renal tubules may be observed in AML, presentation as a cystic mass has not been previously reported. We report the clinicopathologic and immunohistochemical features of four cystic renal AML. The lesions were found in 2 male and 2 female patients, ranging in age from 37 to 76 years, none with a history of hormonal therapy. One of the four patients had known tuberous sclerosis, and this patient and 1 other presented with bilateral cystic renal lesions. Grossly, the lesions measured between 1.3 and 4.5 cm in greatest dimension. Histologically, the lesions were each composed of three components: 1) epithelial cysts lined by cuboidal to hobnail cells; 2) a compact subepithelial "cambium-like" layer of cellular, mullerian-like AML stroma with prominent admixed chronic inflammation; and 3) muscle-predominant AML with associated dysmorphic blood vessels exterior to the cellular subepithelial stroma. Immunohistochemically, the stromal components of all four lesions labeled with HMB-45 and Melan-A, most intensely in the cellular subepithelial stroma, whereas smooth muscle actin and desmin demonstrated the opposite pattern, with greatest intensity in the muscle-predominant AML areas. Immunoreactivity for estrogen and progesterone receptors (ER, PR) and CD10 was present in all cases, with strongest and most diffuse staining in the subepithelial AML cells. The cyst lining cells were positive for pancytokeratin and soy bean agglutinin (the latter characteristic of distal nephron epithelium) but did not label for ER, PR, or melanocytic markers. Although these distinctive lesions may be confused with mixed epithelial-stromal tumor, their lack of female predominance or association with hormone therapy, their distinctive dysplastic blood vessels and disorganized exterior muscular layer, along with their immunohistochemical profile, support their classification as AML with epithelial cysts. Whereas the cysts appear to arise primarily from entrapped collecting ducts, the peculiar subepithelial condensation of AML stromal cells, with morphologic and immunohistochemical features suggesting differentiation toward endometrial stroma, may represent epithelial-induced mullerian differentiation not previously reported in AML. PMID- 16699314 TI - Paraganglioma of the ovary: report of three cases of a rare ovarian neoplasm, including two exhibiting inhibin positivity. AB - Paraganglioma is one of the rarest neoplasms to involve the ovary, whether primary or metastatic, with only two previous reports. We describe three examples that occurred in patients 22, 58, and 68 years of age. Two patients had hypertension. Two tumors involved the left ovary and one the right ovary; they ranged from 8 to 22 cm, were solid, and were tan, brown, or yellow. One tumor was confined to the ovary; in the second case, there were tumor deposits on the posterior surface of the uterus and the contralateral ovary; in the other case, there was peri-aortic lymph node involvement and peritoneal deposits. In all cases, however, radiologic investigations did not reveal an alternative primary site. On microscopic examination, all three tumors showed a predominantly nested "zellballen" pattern with groups of cells surrounded by a vascular stroma. Tumor cells largely had abundant granular eosinophilic cytoplasm with, in 2 cases, focal clear cell areas. In 1 case, bizarre tumor giant cells were present. Immunohistochemically, all neoplasms were cytokeratin negative and diffusely positive with neuroendocrine markers. In 1 case, there was an S-100-positive population of sustentacular cells. Two cases were positive for inhibin, one focal and one diffuse, and the other was focally positive for calretinin. Electron microscopy performed in 2 cases revealed dense core neuroendocrine granules. One patient has been followed up for 15 years and is alive and well. Although metastatic spread from an undetected primary outside the ovary cannot be totally excluded for the 2 cases with extraovarian disease, we think that the neoplasms most likely represent primary ovarian paragangliomas. Because various neoplasms in the sex cord-stromal and steroid categories are likely to enter into the differential diagnosis, inhibin and calretinin positivity represents a significant potential diagnostic pitfall. The differential is broad and may include many other ovarian tumors, particularly those with an oxyphilic cell type. Possible theories of histogenesis of primary ovarian paraganglioma include an origin from extra-adrenal paraganglia in the region of the ovary or unidirectional differentiation within a teratoma. PMID- 16699315 TI - Pathologic findings of lung tumors diagnosed on baseline CT screening. AB - Sixty-five people had a resection of their baseline screen-diagnosed lung cancers in the Early Lung Cancer Action Program. Forty-nine of the carcinomas were solitary, and 42 of these were adenocarcinomas. More than 1 carcinoma was found in 16 patients after pathologic examination of the lobectomy specimen; 15 of the 16 second carcinomas were adenocarcinomas, mixed subtype. Eighteen cases were submitted by local pathologists as Bronchioloalveolar carcinomas but were found to be invasive adenocarcinomas according to the World Health Organization classification by the Pathology Review Panel. Of the 65 resected cases, 57 were N0, 7 were N1, and 1 was N2. Upon careful review of the lobectomy specimens, 49 cases had solitary malignancies, 30 were Stage IA, 13 Stage IB, 3 Stage IIA, 2 Stage IIB, and 1 Stage IIIA on the basis of the American Joint Committee on Cancer/International Union for Cancer Control criteria. In the 16 cases found to have multiple malignancies, 6 had histologically different carcinomas and the remaining 10 had histologically identical malignancies. Eighty-three percent (76/92) of the carcinomas invaded the stroma with destruction of normal lung, and 21% (19/92) also showed either pleural or angiolymphatic invasion, even though 88% (57/65) of the carcinomas were free of lymph node metastases. This report describes the pathologic findings of the resected cases. Histopathologic distinctions among atypical adenomatous hyperplasia, bronchioloalveolar carcinomas, and invasive adenocarcinoma are described in detail. PMID- 16699316 TI - Lymph node involvement in ovarian serous tumors of low malignant potential (borderline tumors): pathology, prognosis, and proposed classification. AB - The occurrence of regional lymph node involvement (LNI) in patients with primary ovarian serous tumors of low malignant potential (S-LMP), although well described in the literature, continues to be problematic. Most studies indicate that LNI is not associated with an adverse prognosis, but there has not been a comprehensive study addressing the histologic patterns of LNI, the importance, if any, of classifying the type of LNI (ie, as either noninvasive or invasive in analogy to peritoneal implant classification), or the presence and significance of associated endosalpingiosis. To further evaluate LNI in S-LMP, 74 patients with ovarian S-LMP and a lymph node biopsy or sampling were studied. Thirty-one of 74 patients had LNI in pelvic (18; 58%), mesenteric/omental (9; 29%), paraaortic (8; 26%), or supradiaphragmatic (2; 6%) lymph nodes. The number of involved nodes ranged from 1 to 20 (mean, 11.1). Four patterns of LNI were identified: individual cells, clusters of cells, and simple, nonbranching papillae (28 of 31; 90%); intraglandular (21 of 31; 68%); cells with prominent cytoplasmic eosinophilia ("eosinophilic cell" pattern) (16 of 31; 52%); and micropapillary pattern (5 of 31; 16%). LNI was diffuse in at least one lymph node in 13 patients (42%) and formed nodular aggregates greater than 1 mm in 6 patients (19%). Nodal endosalpingiosis was present in 58% of cases with LNI compared with 35% without LNI (P=0.06). There was no significant difference in survival for patients with LNI compared with patients without LNI. However, the presence of discrete nodular aggregates of epithelium greater than 1 mm in linear dimension without intervening lymphoid tissue was associated with a statistically significant decreased disease-free survival when compared with other patterns of LNI (P=0.02). Nodular aggregates were strongly associated with desmoplastic fibrous stromal reaction (P=0.001) and micropapillary architecture (0.02). There was also a trend for decreased survival among patients with LNI without associated endosalpingiosis (56%) compared with patients with LNI associated with endosalpingiosis (85%) and those with endosalpingiosis only (93%). This study suggests that patients with ovarian S-LMP may be further substratified into risk categories by the presence of nodular aggregates of S-LMP in lymph nodes, a feature that is more common in cases with micropapillary architecture and associated stromal reaction in the intranodal tumor. This high risk pattern of LNI may have a predictive value similar to invasive peritoneal implants and deserves independent evaluation in future studies of S-LMP. PMID- 16699317 TI - Complex choristoma of the gyrus rectus: a distinct clinicopathologic entity? AB - We report 2 cases of an unusual central nervous system lesion involving the gyrus rectus and characterized by well-differentiated epithelial and mesenchymal components. One patient was a 30-year-old woman, and the other was a 42-year-old man. Both presented with seizures. Radiologically, the lesions were solid, with ill-defined margins and increased signal on T2-weighted MRI. Both were contrast enhancing. There was no significant mass effect or edema. Histopathologically, the epithelial component was characterized by a tubule-forming, stratified epithelium. The lumina of the tubules were lined, in some areas, by a distinct layer of mature keratinocytes. Smooth muscle actin-immunoreactive myoepithelial cells were present in the basal layer, indicating glandular differentiation. Some lumina were cystically dilated and contained anucleate squames and/or PAS positive material. In some areas, the epithelial structures were separated by reactive glial tissue resembling pilocytic astrocytoma, with prominent, piloid processes, Rosenthal fibers, and gemistocytic astrocytes. In other areas, glial nuclei formed small clusters reminiscent of subependymoma, but no glioma was identified in either case. Tissue resembling mature adipose was present focally. No mitotic figures were identified. Follow-up interval of up to 2 years showed no evidence of recurrent tumor. By location and histologic features, this lesion appears to represent a rare, distinct clinicopathologic entity, perhaps derived from misplaced cranial neural crest cells. PMID- 16699318 TI - Schwannoma of the adrenal gland: report of two cases. AB - Primary schwannomas of the adrenal gland are extremely uncommon. We report the clinical and pathologic features of two cases, occurring in a 73-year-old man and a 26-year-old woman, both of whom presented with abdominal pain and an adrenal mass on imaging studies. Both tumors were characterized by a proliferation of cytologically bland spindle cells. One case exhibited alternating compact Antoni A areas and less cellular Antoni B areas consistent with conventional schwannoma, whereas the other case was hypercellular and composed predominantly of Antoni A areas consistent with cellular schwannoma. Reactive inflammatory cells and lymphoid aggregates were present throughout both tumors. The diagnosis of schwannoma was supported by positive immunoreactivity for S-100 protein and collagen IV and absence of reactivity for keratin, muscle related antigens, and CD34 in both cases. Electron microscopy was also performed in the case of cellular schwannoma, which showed ultrastructural features confirming the diagnosis. Both cases had a favorable outcome without evidence of recurrence or metastasis. Because of its rarity, schwannoma occurring at this particular site can pose problems in diagnosis and should be distinguished from other spindle cell lesions of the adrenal gland. PMID- 16699319 TI - Use of chromogenic in situ hybridization to identify MYCN gene copy number in neuroblastoma using routine tissue sections. AB - Amplification of MYCN in neuroblastoma is associated with a poor prognosis. However, methods for estimating the number of MYCN genes based on pooled cells do not address copy number heterogeneity at the cell level and can underestimate or even miss amplification. MYCN copy number can be directly assessed by fluorescence in situ hybridization, but evaluation of tissue histology is next to impossible. We have used a chromogenic method for in situ hybridization (CISH) that enables determination of MYCN copy number using routine light microscopy on routinely processed paraffin sections. Of 41 cases studied, CISH identified 100% of the 18 cases that were determined to be amplified by other techniques and was more sensitive than Southern blotting or quantitative DNA polymerase chain reaction. Because the technique evaluates individual tumor cells, heterogeneity of MYCN copy number was apparent from cell to cell. When defined as 50% or greater variation in copy number between cells in amplified tumors, almost 30% of cases were scored as heterogeneous. Heterogeneity reflects different tumor clones and its role has likely been under-recognized and underestimated in neuroblastoma biology. CISH will provide a valuable tool to assess this phenomenon in conjunction with other morphologic parameters in neuroblastoma specimens, to further our understanding of the biology of this childhood tumor. PMID- 16699320 TI - Fetal pulmonary malformations: defining histopathology. AB - Although classification schemes have sought to categorize congenital cystic lung malformations, studies including the pathology of pulmonary malformations occurring specifically during the fetal period are limited. To better characterize such histopathology, we reviewed a total of 23 fetal lung malformations seen at the Children's Hospital of Philadelphia from 1996 to 2004. Twenty-one of the 23 fetal pulmonary malformations could be categorized into 1 of 3 groups based upon the predominant histologic features present within each lesion. Group 1 (9/21) demonstrated tubular airspaces lined by columnar epithelium. Group 2 (6/21) contained airspaces lined by cuboidal epithelium and surrounded by smooth muscle with abundant interstitial mesenchyme. Group 3 (6/21) showed a mixture of relatively mature-appearing airspaces lined by flattened epithelium and scattered dilated bronchiole-like structures. Cysts were of variable size but in all cases showed a respiratory-type lining. Gestational ages ranged from 21 5/7 to 38 2/7 weeks. Patients in groups 1 and 2 were generally younger than those in group 3; however, morphology did not seem to correlate entirely with normal stages of fetal lung development, and group 2 lesions in particular were the least akin to normal fetal lung. In 4 cases a systemic vascular supply to a lobe of lung was identified, providing evidence that such vasculature is embryonic in origin. The histopathology of fetal lung malformations highlights the variability seen in such lesions at all ages, and it is hoped that continued investigations will provide further insight into these enigmatic lesions. PMID- 16699321 TI - Post-gastric endoscopic mucosal resection surveillance biopsies: evaluation of mucosal changes and recognition of potential mimics of residual adenocarcinoma. AB - Endoscopic mucosal resection (EMR) offers curative treatment for patients with node-negative early gastric carcinoma of less than 2 cm without ulceration or ulceration scar. Follow-up biopsies are frequently performed to ensure the absence of residual neoplasia. We performed a retrospective analysis of post-EMR biopsies from 33 patients who underwent gastric EMR. Histologic changes included inflammation (100%), stromal edema (97.0%), foveolar hyperplasia (78.8%), ectatic vessels (66.7%), epithelial atypia (60.6%), increased glandular mitoses (57.6%), epithelial anisonucleosis (54.5%), fibrinopurulent materials (51.5%), ischemia (48.5%), stromal hemorrhage (33.3%), mucin depletion (12.1%), clear cell degeneration (15.2%), and signet-ring cell-like change (6.1%). Especially, clear cell degeneration and signet-ring cell-like change were conspicuous in the area of ischemia. Residual adenocarcinomas were noted in 4 of 33 cases, and consistently showed high nuclear-to-cytoplasmic ratio with high glandular density. Glandular clear cell degeneration and/or signet-ring cell-like change were worrisome and sometimes difficult to be distinguished from residual neoplastic glands. However, these degenerative glands were usually embedded in a nondesmoplastic stroma and showed anisonucleosis of glandular epithelia. Mimics of residual adenocarcinoma, namely clear cell degeneration and signet-ring cell like change should be judiciously assessed to avoid unnecessary surgery. PMID- 16699322 TI - Assessment and prognostic significance of mitotic index using the mitosis marker phospho-histone H3 in low and intermediate-grade infiltrating astrocytomas. AB - Distinguishing between grade II and grade III diffuse astrocytomas is important both for prognosis and for treatment decision-making. However, current methods for distinguishing between grades based on proliferative potential are suboptimal, making identification of clear cutoffs difficult. In this study, we compared the results from immunohistochemical staining for phospho-histone H3 (pHH3), a specific marker of cells undergoing mitosis, with standard mitotic counts (number of mitoses/10 high-power fields) and MIB-1 labeling index values for assessing proliferative activity. We tested the relationship between pHH3 staining and tumor grade and prognosis in a retrospective series of grade II and III infiltrating astrocytomas from a single institution. The pHH3 index (per 1000 cells), MIB-1 index (per 1000 cells), and number of mitoses per 10 high-power fields were determined for each of 103 cases of grade II and III diffuse astrocytomas from patients with clinical follow-up. pHH3 staining was found to be a simple and reliable method for identifying mitotic figures, allowing a true mitotic index to be determined. The pHH3 mitotic index was significantly associated both with the standard mitotic count and with the MIB-1 index. Univariate analyses revealed that all 3 measurements of proliferation were significantly associated with survival. However, the pHH3 mitotic index accounted for a larger proportion of variability in survival than standard mitotic count or MIB-1/Ki-67 labeling index. After adjusting for age, extent of resection, and performance score, the pHH3 mitotic index remained an independent predictor of survival. Thus, pHH3 staining provides a simple and reliable method for quantifying proliferative potential and for the stratification of patients with diffuse astrocytomas into typical grade II and III groups. These results also suggest that pHH3 staining may be a useful method in other neoplasms in which accurate determination of proliferation potential is relevant to tumor grading or clinical treatment decision-making. PMID- 16699323 TI - Theory versus practice. PMID- 16699324 TI - Delegating decision making in health care organizations. AB - This article explores the realm of delegation from the behavioral standpoint. Delegation is defined as well as reasons why it fails. The limitations of decision making as related to delegation are covered. Ways in which health care organizations can improve managerial decision making to make a positive impact on delegation are explored. Finally, considerations for managers to engage in effective delegation are delineated. PMID- 16699325 TI - Implementing performance management in the Irish Health Sector. AB - To realize the goals of successive health strategies, managers in the Irish Health Sector will have to proactively facilitate optimal employee performance in line with policy objectives. Along with developing employee and teams' capabilities, these managers have begun to implement performance management to achieve the latter. However, there typically are a variety of foundational organizational characteristics required for the successful implementation of performance management. These include providing top-down support for line management buy-in, providing ongoing managerial and performance management training so that trusting relationships and a culture of consensus and cooperation are developed, and appropriately managing expectations. Agreement on employee role definitions and provision of team-based conflict resolution training is also needed to facilitate performance management. There is a need for negotiated performance indicators that are of various types, specific, measurable, and aligned with strategy objectives. Associated reward systems need to be holistic and imaginative, and personal development plans need to have a broader focus than merely improving current job skills and performance. Performance review needs to be ongoing, conducted in a transparent manner, and allocated sufficient discussion time. Managers also need to be mindful of managing overperformance. PMID- 16699326 TI - Job dissatisfaction among public hospital physicians is a universal problem: evidence from Turkey. AB - Several recent reports show that many physicians feel discontented and unhappy with their work situation. This could be due in part to factors such as declining social status, loss of influence on health care organizations, and reduced income. The professional life of physicians is characterized by long working days and pressure for efficiency. This threatens the quality of patient care. In this study we investigated levels of job stress as well as job and life satisfaction among a sample of 168 physicians (80 women and 88 men) who work in a large government facility in Ankara, Turkey. A self-administered questionnaire was distributed to physicians to determine factors contributing to stress and job satisfaction. The most important results of the study show that the participants were unhappy with their salary, did not have enough time to follow developments in medicine, and had a limited social life due to heavy workloads. The results of the study show that physicians who work in public facilities have low motivation due to the nature of their organizations. Public hospital working conditions and staff salaries must be improved to promote health professionals and healthy society. PMID- 16699328 TI - Six Sigma in health care management and strategy. AB - In the years ahead, health care organizations will continue to face numerous challenges from longstanding and currently unresolved issues and new and emerging trends. Some of these include workforce shortages, rising consumerism, patient and stakeholder expectations, quality and patient safety, reimbursement, an aging population, regulatory constraints, and disaster preparedness. Health care organizations will need to adopt effective strategic tools, such as Six Sigma, to improve efficiency and effectiveness. Failure to do so will not only threaten their ability to remain competitive and provide quality care to the communities they serve, but their own survival will also be jeopardized by shrinking margins caused by continued downward pressure on reimbursement. PMID- 16699329 TI - Satisfaction for health care employees: a quest for the Holy Grail? AB - This article proposes that, across industries, too much has been made of the importance of job satisfaction and its impact on organizational effectiveness. In addition, so much attention has been directed toward satisfaction that many health care employees, particularly nurses, now expect job satisfaction from their employers as an entitlement. In nursing, feelings about job satisfaction may, in fact, be exacerbated by the idealism which leads the young person entering the field to expect to be in a "helping profession" where workers almost automatically encounter the satisfaction that comes from giving the help which the patient desperately needs. Faced with the realities of long hours, grueling and often menial tasks, and sometimes churlish patients and physicians, the young nurse may, in fact, enter a period of deep dissatisfaction and of questioning nursing as a career choice. This situation is not unique to nurses. Many health care professionals face equally dissatisfying aspects of their jobs. Our focus is primarily on reducing job dissatisfaction, rather than improving job satisfaction, through practical solutions for those charged with attracting and retaining health care employees during tight labor markets. PMID- 16699330 TI - Can case management interventions reduce the number of emergency department visits by frequent users? AB - This study examined the impact of nurse case management interventions on the number of visits of frequent users of a level 1, urban Emergency Department that sees over 70,000 patient visits per year. Frequent users, defined as those having over 3 visits in a month, were tracked before and after implementation of nurse case management interventions designed to reduce their visit rate. It is a 50 patient pilot study and data collection includes whether or not the patient had a primary care provider, the patient's age and gender, insurance status, and the type of case management interventions including medical social work, community referrals, referrals to primary care providers, and limitation of narcotic prescriptions. Based on statistical tests, pre and post case management interventions suggest that case management interventions do not make a statistically significant reduction in the overall number of visits. This is a medically vulnerable patient group whose visits add to the contemporary problem of Emergency Department overcrowding. The ability of case management interventions to reduce the volume of visits and associated impact on reducing Emergency Department overcrowding was not proven. PMID- 16699331 TI - Shiftwork: what health care managers need to know. AB - Shiftwork is one of health care worker's oldest problems and is known to have important implications on health. Health risks are compounded with age and the amount of cumulated shiftwork. No shift system is clearly advantaged, yet the worker's ability to choose the shift system seems to maximize adaptation to shiftwork. When designing a work schedule, it is important to take into consideration the shift pattern, length of the shift, and the number of consecutive days worked. A poorly designed work schedule can impact the quality of care, the personal and professional outcomes for health care workers, patient satisfaction, length of stay, unplanned absenteeism, cost effectiveness, and productivity. Long-term studies of shiftworkers may disproportionately represent workers who have adapted to shiftwork. Self-scheduling is an interesting alternative in the quest for a more responsive work environment and is a strategy for retention among new, mid-career, and senior nurses. Planned on-site napping may be a useful tool to combat the pernicious effects of sleep debt on performance. Guidelines must be developed and initiatives implemented and evaluated to protect health care workers, especially older female shiftworkers, from the negative impact of shiftwork as they represent a precious resource in a shrinking supply. PMID- 16699332 TI - Health care CIOs: assessing their fit in the organizational hierarchy and their influence on information technology capability. AB - Despite the continuing interest in the importance of health care information technology adoption, very little is known about chief information officers (CIOs), the individuals who manage this effort. The present study surveyed hospital CIOs to understand their backgrounds, their organizational status, and their influence in hospital health care information technology adoption. Survey responses from 98 organizations suggest that the CIO position varies significantly according to the profit status of the hospital. Further, regression analyses suggest that CIO tenure is associated with greater health care information technology adoption, whereas the reporting structure of the CIO is not. Management implications of the findings are discussed. PMID- 16699333 TI - The management of a multigenerational nursing team with differing qualifications: a qualitative study. AB - Retiring age is no longer fixed at 39 years of working service by Italian law, so nurses with more than 40 years of service continue to work in the wards, creating a distinct older generation. Moreover, with the closure of nursing colleges and the introduction of university degree courses for nurses, nursing education has changed significantly. This article reports on a study which explored the experiences of chief nurses in the daily management of nursing teams composed of multigenerational nurses who differed in age and qualifications. Husserls' phenomenologic approach was used, interviewing at length a purposeful sample of 10 chief nurses. Five main areas of concern emerged: (a) The generation gaps between peers and team leaders; (b) nomadism versus being settled; (c) conflicting methods of working which result in nothing being accomplished: managing the "flat paradox"; (d) different nursing values, ideals, and visions; and (e) existing generation gaps with the younger staff. The number of younger nurses will increase with time and, as a consequence, the relevance of intergenerational differences must be seriously debated, not only difficulties with age differences but also those relating to nursing education. PMID- 16699334 TI - Implementation of problem-based learning in a final semester comprehensive nursing course. AB - Problem-based learning is a technique that is becoming increasingly popular in nursing educators in nursing education settings. In the 1980s, Knox and Mogan conducted classic studies on teacher effectiveness as rated by student nurses, graduate nurses, and faculty. They determined that nursing competence and teaching ability were highly rated by both students and faculty to "best" educators. Multiple studies utilizing problem-based teaching styles have revealed that students are highly satisfied with the results of certain characteristics, which promote student learning: self-directed learning, tutor-student interaction, and group learning. This article provides a review of the literature on problem-based learning in nursing education and specifically reviews articles utilizing the Mogan and Knox Nursing Clinical Teacher Effectiveness Inventory. Problems experienced in utilization of problem-based learning are also described. PMID- 16699336 TI - National Nurse Month. PMID- 16699338 TI - Advancing the agenda for home healthcare quality: conference proceedings and findings. AB - On March 31 to April 1, 2005, the Center for Home Care Policy and Research of the Visiting Nurse Service of New York assembled approximately 80 home healthcare stakeholders from across the country to advance the action strategy for measuring, improving, and assuring quality in home healthcare. A specific aim of the conference was to disseminate relevant research on organizational change and evidence-based practices to influence patient safety and quality of care. Attendees worked together to identify high-priority action steps and recommendations at both the organizational and policy levels. This article reports the conference's proceedings and findings. PMID- 16699339 TI - Diabetes management at the end of life: transitioning from tight glycemic control to comfort. AB - Tight glycemic control has become the standard of care for prevention of the long term side effects of diabetes mellitus. When individuals with diabetes approach the end of life from advanced cancer or another chronic illness, they often become anorexic. The result is an increased risk for hypoglycemic episodes. It is appropriate to shift the goal of therapy from tight control of blood sugar to maintaining comfort and enhancing quality of life. PMID- 16699340 TI - Honoring our elders: hearing their stories, respecting their ways. PMID- 16699341 TI - Medical devices: promoting a safe migration into the home. AB - Medical device use in the home environment has grown in volume and complexity. Because of this, the Food and Drug Administration (FDA) needs to ensure that these products are safe and effective for use in this environment. The FDA must develop a partnership with home healthcare providers in areas of patient education, monitoring device adverse events that occur in the home environment, and communicating problems to the manufacturer. PMID- 16699343 TI - Using standards of care to drive evidence-based clinical practice and outcomes for diabetes mellitus. AB - Home care has the opportunity to positively affect the quality of care and outcomes of patients with diabetes by implementing evidence-based practice. However, many clinicians are unsure how to locate and use this guidance. This article describes how the staff nurse can use evidence-based practice interventions and outcomes from the American Diabetes Association's standards to provide optimal care for patients with diabetes. PMID- 16699345 TI - What home healthcare nurses are saying about their jobs. PMID- 16699346 TI - Foot care: prevention of problems for optimal health. AB - Problems of the feet generally cause pain but are not life threatening. By making information available regarding ill-fitting shoes and the conditions linked to poor shoe wear, the awareness of our clients can be raised. Home health nurses can contribute to the educational process that may reduce foot problems associated with improper shoe wear. As the foot ages, the normal plantar fat pad begins to atrophy. During the initial visit, foot health should be integrated into health promotion. This article seeks to provide an overview of common foot problems that are not related to diabetes mellitus. PMID- 16699350 TI - Sister Jeanne. PMID- 16699351 TI - Considerations in prevention of surgical site infections following cardiac surgery: when your patient is diabetic. AB - The incidence of surgical site infections among patients with diabetes continues to occur at a greater rate when compared with their nondiabetic counterparts. Preexisting vascular changes, delayed wound healing, and impaired immune factors contribute. Adult patients with diabetes likely possess comorbid coronary artery disease, thus increasing the need for cardiac surgery. The resultant potential for infection can be combated with supplementary interventions above those universally taken. Modifiable risk factors of hyperglycemia and obesity are targeted preoperatively. Glycemic control, adequate tissue perfusion, and adjunct use of nasal mupirocin are addressed intraoperatively. Lastly, focus is placed on nutrition, exercise, and continued glucose control postoperatively and beyond discharge. PMID- 16699352 TI - ALOHA to women's heart disease. AB - This article describes the American Heart Association, ALOHA program. ALOHA is a multidisciplinary approach to helping lay people and clinicians determine the best course of action for managing cardiac risk factors in women. ALOHA, an acronym that stands for designated interventions based on individualized assessment of patients, along with the Framingham risk assessment calculator, allows health care providers with their patients to individualize treatment for heart disease and management of risk factors. PMID- 16699354 TI - An introduction to the collaborative methodology and its potential use for the management of heart failure. AB - BACKGROUND: Heart failure (HF) is responsible for significant disease burden in developed countries internationally. Despite significant advances and a strong evidence base in therapies and treatment strategies for HF, access to these therapies continues to remain elusive to a significant proportion of the HF population. The reasons for this are multifactorial and range from the financial cost of treatments to the individual attitudes and beliefs of clinicians. The collaborative methodology, based upon a quality improvement philosophy, has been identified as a potentially useful tool to address this treatment gap. AIM: In this manuscript, we review the published literature on the collaborative methodology and assess the evidence for achieving improvement in the management of HF. METHODS: Searches of electronic databases, the reference lists of published materials, policy documents, and the Internet were conducted using key words including "collaborative methodology," "breakthrough series," "quality improvement," "total quality improvement," and "heart failure." Because of the paucity of high-level evidence, all English-language articles were included in the review. RESULTS: On the basis of the identified search strategy, 43 articles were retrieved. Key themes that emerged from the literature included the following: (1) The collaborative methodology has a significant potential to reduce the treatment gap. (2) Leadership is an important characteristic of the collaborative method. (3) The collaborative methodology facilitates sustainability of the quality improvement process. CONCLUSION: The collaborative methodology, when implemented and conducted according to key conceptual principles, has significant potential to improve the outcomes of patients, particularly those with HF and chronic cardiovascular disease. PMID- 16699355 TI - Factors influencing patient knowledge of warfarin therapy after mechanical heart valve replacement. AB - BACKGROUND AND RESEARCH OBJECTIVE: Patients with mechanical heart valves must follow lifelong warfarin therapy. Warfarin, however, is a difficult drug to manage because it has a narrow therapeutic window and potentially serious side effects. Successful anticoagulation treatment is dependent upon the patient's knowledge of this drug; however, little is known regarding the determinants of such knowledge. Therefore, the purpose of this study was to determine the influence of both in-hospital teaching practices as well as socioeconomic status and demographic variables on patients' knowledge of warfarin therapy. SUBJECTS AND METHODS: A telephone survey was conducted among 100 patients 3 to 6 months after mechanical heart valve replacement. A previously validated 20-item questionnaire was used to measure the patient's knowledge of warfarin, its side effects, and vitamin K food sources. Demographic information, socioeconomic status data, and medical education information were also collected. Knowledge scores were compared using the Student t test or one-way analysis of variance. Variables with P < or = .2 on univariate analysis were entered in multiple stepwise regression analysis. RESULTS AND CONCLUSIONS: Sixty-one percent of participants had scores indicative of insufficient knowledge of warfarin therapy (score < or = 80%). Age was negatively related to warfarin knowledge scores (r = 0.27, P = .007). Patients with family incomes greater than $25,000, who had greater than a grade 8 education, and who were employed or self-employed had significantly higher warfarin knowledge scores (P = .007, P = .002, and P = .001, respectively). Gender, ethnicity, and warfarin therapy before surgery were not related to warfarin knowledge scores. Furthermore, none of the in-hospital teaching practices significantly influenced knowledge scores, whereas receiving postdischarge community counseling significantly improved knowledge scores (P = .001). Multivariate regression analysis revealed that understanding the concept of International Normalized Ratio, knowing the acronym, age, and receiving community counseling after discharge were the strongest predictors of warfarin knowledge. Accessing postdischarge counseling resulted in significantly improved warfarin knowledge scores. Because improved knowledge has been associated with improved compliance and control, our findings support the need to develop a comprehensive postdischarge education program or at least to ensure that patients have access to a community counselor to compliment the in-hospital education program. PMID- 16699357 TI - Depression, physical impairment, and treatment of depression in chronic heart failure. AB - BACKGROUND AND RESEARCH OBJECTIVE: This study examines the contribution of attitudes about impairment to the relation between depression and physical impairment in patients with heart failure. It also describes the current status of antidepressant treatment in a sample of outpatients with heart failure. SUBJECTS AND METHODS: A total of 32 depressed and 51 nondepressed patients with heart failure were recruited while seeking heart failure treatment in an outpatient heart failure or family practice clinic. Medical, functional, cognitive, and psychological measures were administered at baseline. Depression measures were readministered to the 32 depressed patients at 8, 16, and 24 weeks after the baseline interview. RESULTS AND CONCLUSIONS: Attitudes about impairment and perceived social support were the strongest cross-sectional correlates of depression. The strong association between physical impairment and depression was no longer significant after controlling for attitudes about impairment. In addition, attitudes about impairment predicted chronicity of depressive symptoms longitudinally after controlling for baseline depressive symptom severity. The relation between physical impairment and depression in heart failure is strongly related to how patients cope with impairment. These results call for the development of additional interventions to treat depression that focus on the emotional adjustment to physical impairment. PMID- 16699358 TI - A nursing intervention to reduce prehospital delay in acute coronary syndrome: a randomized clinical trial. AB - Prehospital delay in patients experiencing symptoms of acute coronary syndrome (ACS) has proved to be a significant and intractable public health problem, with minimal change in delay time over the past 2 decades despite numerous community education programs designed to reduce delay. With each 30-minute increment of delay, 1-year mortality increases by 7.5%, thus reinforcing the importance of helping patients label symptoms correctly and take appropriate action steps to seek definitive treatment. We therefore are conducting a multicenter, international clinical trial in 3,500 patients with documented coronary heart disease to determine whether a brief education and counseling intervention delivered by a nurse can reduce prehospital delay in the face of symptoms of ACS. The main outcome being studied is time from ACS symptom onset to arrival at the emergency department. Secondary outcomes include use of the emergency medical system; aspirin use; and knowledge, attitudes, and beliefs about heart attack symptoms. Patients are being followed for 2 years from the time of enrollment. The purpose of this article is to describe the intervention and its theoretical framework, and to outline the design of this randomized controlled trial. PMID- 16699359 TI - Effects of music therapy on physiological and psychological outcomes for patients undergoing cardiac surgery. AB - BACKGROUND: Cardiac surgery is a common interventional procedure for ischemic and valvular heart disease. Cardiac surgery is accompanied by postoperative pain and anxiety. The use of music therapy has been shown to reduce pain, anxiety, and physiological parameters in patients having surgical procedures. OBJECTIVES: To compare the effects of music therapy versus a quiet, uninterrupted rest period on pain intensity, anxiety, physiological parameters, and opioid consumption after cardiac surgery. SUBJECTS AND METHODS: An experimental design was used. A total sample of 86 patients (69.8% males) were randomized to 1 of 2 groups; 50 patients received 20 minutes of music (intervention), whereas 36 patients had 20 minutes of rest in bed (control). Anxiety, pain, physiologic parameters, and opioid consumption were measured before and after the 20-minute period. RESULTS: A significant reduction in anxiety (P < or = .001) and pain (P = .009) was demonstrated in the group that received music compared with the control group, but no difference was observed in systolic blood pressure (P = .17), diastolic blood pressure (P = .11), or heart rate (P = .76). There was no reduction in opioid usage in the 2 groups. CONCLUSIONS: Patients recovering from cardiac surgery may benefit from music therapy. PMID- 16699360 TI - Adherence to treatment factors in hypertensive African American women. AB - Hypertension (HTN) is a major public health problem in African American women. The disproportionate rate of HTN in African American women makes it a high priority national health objective. Lack of access to a regular continuous source of care may be particularly significant among African American women because such a source of care is a prerequisite to early diagnosis and successful treatment. Understanding the factors that influence access to care for HTN in African American women will provide the information needed to improve the use of healthcare services and enhance adherence to treatment. Despite the known importance of adherence to successful HTN treatment, few attempts have been made to incorporate the indicators of adherence to HTN treatment into a theoretical model that can be used to guide research and practice with African American women. The adapted Hill-Levine model offers an appropriate framework for studying these phenomena in African American women because factors frequently associated with seeking and using healthcare services are organized in three dimensions, including predisposing, enabling, and reinforcing factors. This article discusses the magnitude of the problem presented by HTN and lack of HTN control in African American women, identifies from the literature the factors related to adherence to treatment in HTN management, in general, and in African American women, specifically, and uses the adapted Hill-Levine model to organize the identified predisposing, enabling, and reinforcing factors associated with adherence to HTN treatment recommendations among African American women. PMID- 16699361 TI - Recruiting and retaining young, sedentary, hypertension-prone African American women in a physical activity intervention study. AB - African American women have a high prevalence of hypertension and low level of physical activity compared with their counterparts. A sedentary lifestyle contributes to the development of hypertension, as well as other cardiovascular diseases, especially among African American women. Healthy People 2010 initiatives underscore the priority of reducing minority health disparities. To reduce health disparities, there has been recent emphasis on recruiting and retaining minority populations in clinical research studies. However, little information is available to guide researchers in the evaluation of impediments in successful recruitment and retention of young African American women. A first step is for researchers to report information concerning the efficacy of recruiting/retaining methods in order to facilitate minority participation in clinical trials and, ultimately, reduce health disparities. This report summarizes existing recruitment and retention methods from the literature, and describes how effective these strategies were in recruiting and retaining young, mildly hypertensive African American women to a physical activity intervention study. Multiple strategies, resources, and time were necessary to recruit and retain these women for the study. Among women enrolled, newspaper advertisements and flyers were the most effective recruiting strategies implemented (46% and 21%, respectively). Study retention was high (96%), which may have resulted from flexible scheduling, frequent contact, and a caring environment. Recruiting and retaining efforts need to be tailored to meet the needs of the target population. PMID- 16699362 TI - Opportunistic screening for cardiovascular problems in rural and remote health settings. AB - BACKGROUND: Cardiovascular disease is the leading cause of death in Australia and the United States. It is not known if routine electrocardiogram (ECG) assessment at the community level could identify a significant proportion of people with clinically relevant ECG anomalies who could benefit from intervention. PURPOSE: This study aimed to elucidate the use of 3-lead ECG assessment by community nurses in rural and remote health settings. We report the findings obtained from 20-minute, lead II ECG recordings of 71 people who participated in a diabetes screening study. RESULTS: Seven participants reported cardiac anomalies before screening. One or more ECG abnormalities were found in 45 participants. Of these, nine people who were otherwise asymptomatic showed abnormal ECG characteristics that warranted further investigation. CONCLUSION: Although further research is needed, incorporating routine 3-lead ECG testing in rural and remote communities may improve general health in the community by providing early recognition of cardiac anomalies in otherwise asymptomatic individuals who may be amenable to treatment. This study has implications for community nurses on two levels: opportunistic screening, monitoring and evaluation. PMID- 16699363 TI - End-of-life communication and hospital nurses: an educational pilot. AB - Heart failure is a high prevalence, high burden disease with an unpredictable trajectory. Given that approximately 50% of persons with the diagnosis die within 5 years, the implications for communication about advance planning and end of life, although simultaneously providing hope and treatment, are extremely challenging. This article reports on a feasibility trial of a modest continuing education seminar to increase awareness and communication skills of inpatient nurses. Curriculum design details, including objectives and methods, are described. Needs assessment and evaluation data are reported. At the 2-month posttest, a summary communications skill score showed significant improvement. Data also illustrated that the challenge of facing issues about dying are complicated by a work force of relatively young staff and low exposure to few actual deaths during relatively short lengths of stay. PMID- 16699364 TI - A motivational counseling approach to improving heart failure self-care: mechanisms of effectiveness. AB - BACKGROUND: Self-care is an integral component of successful heart failure (HF) management. Engaging patients in self-care can be challenging. METHODS: Fifteen patients with HF enrolled during hospitalization received a motivational intervention designed to improve HF self-care. A mixed method, pretest posttest design was used to evaluate the proportion of patients in whom the intervention was beneficial and the mechanism of effectiveness. Participants received, on average, 3.0 +/- 1.5 home visits (median 3, mode 3, range 1-6) over a three-month period from an advanced practice nurse trained in motivational interviewing and family counseling. Quantitative and qualitative data were used to judge individual patients in whom the intervention produced a clinically significant improvement in HF self-care. Audiotaped intervention sessions were analyzed using qualitative methods to assess the mechanism of intervention effectiveness. RESULTS: Congruence between quantitative and qualitative judgments of improved self-care revealed that 71.4% of participants improved in self-care after receiving the intervention. Analysis of transcribed intervention sessions revealed themes of 1) communication (reflective listening, empathy); 2) making it fit (acknowledging cultural beliefs, overcoming barriers and constraints, negotiating an action plan); and, 3) bridging the transition from hospital to home (providing information, building skills, activating support resources). CONCLUSION: An intervention that incorporates the core elements of motivational interviewing may be effective in improving HF self-care, but further research is needed. PMID- 16699365 TI - Progress in prevention: adding a cerebral dimension. PMID- 16699366 TI - Immune suppression in the tumor microenvironment. AB - The identification of tumor-expressed antigens that can be recognized by specific T lymphocytes has made it possible both to study the properties of T cells participating in anti-tumor immune responses in patients and also to develop antigen-specific immunotherapies as a treatment modality. Interestingly, moves toward intervention have proceeded at a faster pace than have investigations toward understanding. In melanoma in particular, many clinical trials of active immunization have been performed, and many of these have shown increases in tumor antigen-specific T cells circulating in the blood. However, clinical responses have been infrequent, arguing that mechanisms of resistance downstream from initial T cell priming may be dominant in many cases. In fact, may patients show spontaneous generation of immune effector cells and/or antibodies, implying that the priming phase has occurred already in such individuals even without vaccination. Recent attention has turned toward mechanisms of immune evasion at the effector phase of the anti-tumor immune response, predominantly within the tumor microenvironment. Evidence is accumulating that T cell-intrinsic hyporesponsiveness or anergy, extrinsic suppression by regulatory cell populations, inhibitory ligands such as PD-L1, soluble factors such as TGF-beta, and the activity of nutrient-catabolizing enzymes such as indoleamine 2,3 dioxygenase (IDO), may contribute to immune escape in different settings. Murine preclinical models have shown that interfering with each of these processes can translate into T cell-mediated tumor control. Clinical studies to estimate the frequency of specific immune evasion mechanisms in individual patients, to correlate specific events with clinical outcome, and to develop strategies to counter resistance mechanisms should receive a high priority. PMID- 16699367 TI - Accumulation in tumor tissue of adoptively transferred T cells: A comparison between intravenous and intraperitoneal injection. AB - Accumulation of T cells at the tumor is essential in cancer immunotherapy based on adoptive transfer of tumor-specific T cells. To gain further insight into the accumulation process and to evaluate the effect of using different routes of cell transfer, we investigated the accumulation of ovalbumin-specific CD8+ T cells (OT I) injected either intravenously (IV) or intraperitoneally (IP) into mice carrying a subcutaneous tumor of the ovalbumin-expressing melanoma cell line B16 OVA. Maximal accumulation of the adoptively transferred cells in tumor tissue was observed 5 days after injection, irrespective of the injection route. The route of injection affected neither the total number of adoptively transferred cells found in tumor tissue nor the kinetics of this accumulation. In the spleen, however, the accumulation of adoptively transferred cells was clearly dependent on the injection route. IP injections resulted in a large number of adoptively transferred cells in the spleen on all days analyzed. In comparison, IV injection resulted in significantly fewer adoptively transferred cells in the spleen, and this number decreased over time. The route of injection affected neither the activation status of the adoptively transferred T cells that accumulated at the tumor site, nor the ability of these cells to control tumor growth. Two cell populations, SIINFEKL-tetramer(Low)(Tet(Low))CD69+ CD25+ and Tet(high)CD69- CD25 , were present in tumor samples, whereas only Tet(High)CD69- CD25- cells accumulated in the spleen. In tumors, IV injection resulted in a higher fraction of adoptively transferred cells with an activated phenotype (Tet(Low)CD69+ CD25+) compared with IP injection. PMID- 16699368 TI - Assessing the safety of cytotoxic T lymphocytes transduced with a dominant negative transforming growth factor-beta receptor. AB - Transforming growth factor (TGF)-beta, a pleiotropic cytokine that regulates cell growth, is secreted by many human tumors and markedly inhibits tumor-specific cellular immunity. It has previously been shown by our group that transduction of cytotoxic T lymphocytes (CTLs) with a retroviral vector expressing the dominant negative TGFbeta type II receptor (DNR) overcomes this tumor evasion in a model of Epstein-Barr virus (EBV)-positive Hodgkin disease. TGFbeta is an important physiologic regulator of T-cell growth and survival, however, abrogation of this regulatory signal in genetically modified cells is potentially problematic. To ensure that unresponsiveness to TGFbeta did not lead to the unregulated growth of genetically modified CTLs, the characteristics of DNR-transduced CTLs in vivo were studied. Donor C57BL6 mice were vaccinated with human papillomavirus-E7 plasmid DNA to induce production of E7-specific CTLs. The E7-specific CTLs were genetically modified to express enhanced green fluorescent protein (GFP) or DNR and administered to syngeneic mice. All mice received monthly boosts with E7 DNA for 9 months, and during this time, transduced CTLs were detected in the peripheral blood of most of the mice using a quantitative real-time polymerase chain reaction. By 12 months, 3 months after cessation of vaccination, no DNR transduced CTLs or GFP-transduced CTLs were detected in the peripheral blood. There were 4 cases of lymphoma (2 DNR-transduced mice and 2 control mice): all tumors were CD3-/CD8- and were also negative for the DNR transgene. Hence, mature antigen-specific cytotoxic T cells can be genetically modified to resist the antiproliferative effects of TGFbeta without undergoing spontaneous lymphoproliferation in vivo. They may be of value for treating human cancers, which use TGFbeta as a powerful immune evasion mechanism. PMID- 16699369 TI - Enhanced immunity to breast cancer in mice immunized with fibroblasts transfected with a complementary DNA expression library from breast cancer cells: Enrichment of the vaccine for immunotherapeutic cells. AB - Breast cancer cells express an array of weakly immunogenic tumor-associated antigens (TAAs). Under appropriate circumstances, immunity to breast cancer can be induced, with potential benefits for patients with the disease. Here, we report a new cell-based vaccination strategy resulting in enhanced immunity to breast cancer in tumor-bearing mice. The strategy was designed to enrich the vaccine for highly immunogenic cells. The vaccine was prepared by transfer of unfractionated complementary DNA (cDNA) derived from a highly malignant breast neoplasm that arose spontaneously in a C3H/He mouse (SB5b) into an immunogenic fibroblast cell line. As the transferred cDNA spontaneously integrates into the genome of the recipient cells and is replicated as the cells divide, sufficient DNA to prepare the vaccine could be obtained from as few as 10(7) cells (4-mm tumor). Because only a small proportion of the transfected cell population was expected to have incorporated genes responsible for inducing immunity to the breast cancer, we devised a novel approach designed to enrich the transfected cell population for cells that induced immunity to the neoplasm. Aliquots of the transfected population were divided into small pools (initial inoculum = 4 x 10). Afterward, the cell number from each pool was allowed to expand in vitro. Pools containing greater numbers of immunogenic cells (identified by 2 independent assays) were subdivided for additional rounds of immune selection. Enhanced immunity to the neoplasm was detected in tumor-bearing mice treated solely by immunization with the enriched cell population. The immunity, mediated by CD8+ T cells, was sufficient to prolong the survival of mice with established breast cancer. PMID- 16699370 TI - H60/TNT-3 fusion protein activates NK cells in vitro and improves immunotherapeutic outcome in murine syngeneic tumor models. AB - H60 is a murine minor histocompatibility antigen that binds to NKG2D and activates an effector phenotype in NK and T cells. In the present study, H60 was genetically fused to the tumor-targeting murine MAb TNT-3. The resultant fusion protein, named H60/TNT-3, was produced in NS0 cells and determined by ELISA to possess an H60 epitope. The Ka of H60/TNT-3 (2.43 x 10(9) M(-1)) was nearly identical to that of the parental Ab (2.22 x 10(9) M(-1)), demonstrating that addition of the H60 moiety to the N-terminus of TNT-3 heavy chain did not affect antigen affinity. In vitro, H60/TNT-3 bound and activated murine NK cells, eliciting IFN-gamma production in a higher percentage of cells than the activating NKG2D Ab A10. In vivo, H60/TNT-3 possessed a half-life of approximately 12 hours and effectively targeted tumor tissue versus control organs, with nearly 2% injected dose per gram of tumor retained after 48 hours. Finally, H60/TNT-3 was tested for antitumor efficacy in BALB/c and C57BL/6 mice bearing subcutaneous syngeneic carcinomas. Tumor volume reduction was observed in both CT26 and Lewis Lung models (53% and 52%, respectively) relative to untreated control mice. Further, Lewis Lung carcinoma-bearing mice treated with H60/TNT-3 experienced a statistically significant survival advantage. Taken together, these data characterize a new immunotherapeutic MAb with antitumor efficacy that prolonged overall survival in a resistant solid tumor model. PMID- 16699371 TI - Comparison of common gamma-chain cytokines, interleukin-2, interleukin-7, and interleukin-15 for the in vitro generation of human tumor-reactive T lymphocytes for adoptive cell transfer therapy. AB - The adoptive transfer of human tumor-reactive T lymphocytes into autologous patients can mediate the regression of metastatic melanoma. Here, the in vitro generation of melanoma-reactive T lymphocytes was compared using 3 common gamma chain cytokines, interleukin (IL)-2, IL-7, and IL-15, alone or in combination. The proliferation, function, and phenotype were evaluated for tumor-reactive T cells derived from peripheral blood mononuclear cells (PBMCs) from patients previously immunized with the melanoma-associated peptide gp100:209-217(210M) and PBMCs transduced with a retrovirus encoding the alpha and beta chains of a gp100 reactive T-cell receptor (TCR). IL-7 alone did not induce significant proliferation of any tumor-reactive T-cell population, whereas IL-2 and IL-15 induced significant proliferation of tumor-reactive T lymphocytes from both sources. Cells cultured in the presence of IL-2 or IL-15 secreted comparable amounts of interferon-gamma and IL-2 in response to melanoma cells in vitro and were phenotypically similar in terms of costimulatory molecules (CD27 and CD28), cytokine receptors (CD25, CD122, and CD127), and a lymphoid homing molecule (CD62L). In addition, the proliferation, function, and phenotype of T cells cultured with combinations of IL-2, IL-7, and IL-15 were similar to those grown with IL-2 alone. The effects of these cytokines on TCR stimulation of CD45RA+ naive cells derived from adult patients and from human umbilical cord blood were also compared. Similar to the data with activated tumor-reactive T lymphocytes, IL-7 alone did not support significant proliferation of naive T cells after TCR stimulation with anti-CD3, although IL-2 and IL-15 induced comparable proliferation of T lymphocytes with similar phenotypic attributes. PMID- 16699372 TI - Cytotoxic T lymphocytes responding to low dose TRP2 antigen are induced against B16 melanoma by liposome-encapsulated TRP2 peptide and CpG DNA adjuvant. AB - The induction of a potent and specific T cell response is a major challenge in the development of efficacious cancer vaccine strategies. We applied a novel liposomal formulation (AVE3) for efficient delivery of antigenic peptides into APCs of the skin. These liposomes resulted in a long-lasting deposition of encapsulated compounds at the injection site and the draining lymph nodes. Using a peptide from the melanocyte differentiation antigen tyrosinase-related protein (TRP2) 2 we could show that vaccination with liposome-encapsulated peptide in combination with oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODNs) as adjuvant leads to the induction of tumor cell-specific cytotoxic T cells. The most potent immune response was observed when both, TRP2 peptide and CpG ODNs, were encapsulated into AVE3. Importantly, in contrast to vaccination with free TRP2 liposomal TRP2 peptide generated T cells which respond to 1000 fold lower antigen concentration. Using the poorly immunogenic B16 melanoma model we could demonstrate that vaccination with liposomal TRP2 peptide plus CpG ODNs but not vaccination with free peptide or adjuvant alone resulted in tumor protection in subcutaneous and metastatic tumor models. In summary, vaccination with liposome-encapsulated peptide antigen and CpG ODN allows for the in vivo loading and activation of DC, thereby generating reactive CTL populations even against poorly immunogenic self-peptide presenting tumors resulting in a potent anti-tumor immune response. PMID- 16699373 TI - Myelopeptide-2 recovers interleukin-2 synthesis and interleukin-2 receptor expression in human T lymphocytes depressed by tumor products or measles virus. AB - Myelopeptide-2 (MP-2; Leu-Val-Val-Tyr-Pro-Trp), originally isolated from the supernatant of porcine bone marrow cell culture, is able to restore the mitogen responsiveness of human T lymphocytes inhibited by conditioned medium from HL-60 leukemia cells or measles virus. This effect is based on the ability of MP-2 to recover the reduced interleukin (IL)-2 synthesis and IL-2 receptor (IL-2R) expression in human T lymphocytes treated with these harmful agents. The involvement of other cytokines in MP-2 restoration of the reduced IL-2 synthesis in T lymphocytes is experimentally studied. It is shown that T helper (TH) 1 and TH2 cytokines are acting in close interaction, the character of which depends on the immune status of the T-lymphocyte donors. The data obtained allow one to suggest that the MP-2 involvement in regulatory processes is directed to the maintenance of immune homeostasis. This peptide is perspective to be applied in antitumor and antivirus therapy. PMID- 16699374 TI - IL-7 administration to humans leads to expansion of CD8+ and CD4+ cells but a relative decrease of CD4+ T-regulatory cells. AB - Lymphopenia is a serious consequence of HIV infection and the administration of cancer chemotherapeutic agents. Although growth factors can be administered to patients to increase circulating neutrophils, there is no effective method to stimulate CD8+ lymphocyte production in humans, in vivo. This report is the first to describe the administration of recombinant interleukin-7 to humans and demonstrates the ability of this cytokine to mediate selective increases in CD4+ and CD8+ lymphocytes along with a decrease in the percentage of CD4+ T-regulatory cells. These studies suggest an important role for interleukin-7 in the treatment of patients with lymphopenia. PMID- 16699375 TI - Efficient ex vivo expansion of Valpha24+ NKT cells derived from G-CSF-mobilized blood cells. AB - Natural killer T (NKT) cells are involved in the function of innate immune systems and also play an important role in regulating acquired immune responses. In previous reports, we showed that Valpha24+ NKT cells proliferated more efficiently from granulocyte-colony stimulating factor (G-CSF)-mobilized peripheral blood mononuclear cells (PBMC) than from non-mobilized PBMC. However, the mechanism of this enhanced NKT cell expansion is not yet clear. The goal of this research was to develop culture conditions for the more efficient ex vivo expansion of NKT cells. G-CSF-mobilized PBMC was cultured in AIM-V medium supplemented with 10% auto-plasma, 100 ng/mL alpha-galactosylceramide (alpha GalCer) and 100 IU/mL recombinant human (rh) interleukin (IL)-2. The efficiency of the expansion of Valpha24+ NKT cells was evaluated on day 12. The expansion fold of Valpha24+ NKT cells was augmented depending on the proportion of CD14+ cells at the beginning of culture. The depletion of Valpha24+ NKT cells abrogated the expansion of Valpha24+ NKT cells. Depletion of CD56+ NK cells from mobilized PBMC enhanced, and add-back of purified CD56+ NK cells suppressed the expansion of Valpha24+ NKT cells. Experiments with different timings for the addition of cells, IL-2 and alpha-GalCer suggested that follow-up supplementation with IL-2 or CD14+ cells should be avoided for the efficient expansion of Valpha24+ NKT cells. These results should be useful for the development of an efficient and practical expansion protocol for adoptive immunotherapy with Valpha24+ NKT cells. PMID- 16699376 TI - Application of the granzyme B ELISPOT assay for monitoring cancer vaccine trials. AB - Granzyme B (GrB) is present in the granules of cytolytic lymphocytes and is a key mediator of cell-mediated target cell death via the granule-mediated pathway. The release of GrB can be used as an indicator of a cytotoxic T lymphocyte response. Herein, we report that the GrB enzyme-linked immunospot assay (ELISPOT) can be used to measure ex vivo antigen-specific cytotoxicity of peripheral blood mononuclear cells from cancer patients vaccinated with a peptide-based cancer vaccine. We compare the reactivity of patients' peripheral blood mononuclear cells in the GrB ELISPOT with reactivity in the tetramer, interferon (IFN)-gamma ELISPOT, and Cr-release assays. Differences in immune response over all assays tested were found between patients and 4 response patterns were observed. Reactivity in the GrB ELISPOT was more closely associated with cytotoxicity in the Cr-release assay than the tetramer or IFN-gamma ELISPOT assays. Moreover, the higher affinity g209-2M peptide (used for vaccination) elicited greater GrB secretion than the native g209 peptide, although this difference was not observed with IFN-gamma secretion. Taken together with the fact that GrB is a specific mediator released by cytotoxic T lymphocytes, these results show that simultaneous use of the GrB ELISPOT assay with other immunologic assays may provide important additional immunologic insight into patient responses to cancer vaccines. PMID- 16699377 TI - Highly efficient expansion of human CD4+CD25+ regulatory T cells for cellular immunotherapy in patients with graft-versus-host disease. AB - CD4+CD25+ regulatory T cells (T(REG)) are engaged in the regulation of murine and human immune responses as well as graft-versus-host disease (GvHD) after allogeneic stem-cell transplantation. Despite their suppression of GvHD they do not impair graft-versus-tumor activity in the mouse, which makes T(REG) especially attractive candidates for cellular immunotherapy. T(REG) comprise only 5% to 10% of CD4+ T cells in peripheral blood and are naturally anergic, which prevented their use as therapeutic suppressor cells in the context of autoimmune or alloimmune reactions so far. We therefore developed an in vitro expansion protocol for human T(REG), breaking their anergy with anti-CD3/anti-CD28-coupled paramagnetic beads and a combination of interleukin (IL)-2 and IL-15. Highly purified human T(REG) can be expanded 285-fold to 1000-fold within 20 days and keep their phenotype as well as all their suppressor functions even in the context of stimulation with mature allogeneic dendritic cells. However, we demonstrate that FoxP3 is not a reliable marker for human T(REG) as it is transiently inducible in CD4+CD25- cells upon activation with cytokines or via their T cell receptor. In addition, we successfully expanded CD4+CD25+ cells from patients after allogeneic stem-cell transplantation with or without GvHD and show that different suppressor functions might be lost independently, demonstrating that human T(REG) biology is likely more complicated than previously thought. PMID- 16699378 TI - Diagnosing major depressive disorder III: can some symptoms be eliminated from the diagnostic criteria? AB - All criteria used to diagnose a psychiatric disorder should contribute to distinguishing cases from noncases. The principal of parsimony argues for defining a disorder with as few criteria as possible. Thus, criteria that do not contribute to the case-noncase distinction should be eliminated from the list of defining features because they unnecessarily increase the complexity of the definition of the disorder. In polythetically defined disorders such as major depressive disorder (MDD), diagnosis is based on the presence of a minimum number of features from a list. For a criterion to be retained on such a list, it should contribute to distinguishing between individuals with and without MDD. Simply demonstrating that a criterion is significantly more common in individuals with MDD than individuals without MDD is not a sufficient demonstration of its necessity. Rather, to demonstrate an impact on diagnosis, it should be shown that eliminating the criterion from the list results in individuals being reclassified from a case to a noncase. A criterion does not contribute to determining caseness if its elimination does not result in diagnostic reclassification. The goal of this report from the Rhode Island Hospital Methods to Improve Diagnostic Assessment and Services project was to determine if any of the criteria of MDD are candidates for elimination because of their lack of impact on diagnosis. The results indicated that the symptoms of increased weight, decreased weight, and indecisiveness rarely influenced diagnostic classification and thus are candidates for elimination. PMID- 16699379 TI - Major depression in the transition to adulthood: the impact of active and past depression on young adult functioning. AB - This study examined the association between active and past major depression and deficits in young adult functioning using data from a longitudinal community study (N = 354). Three groups were compared: (1) participants with a 1-year diagnosis of major depression at age 26 (active group); (2) those who experienced major depression during the transition to adulthood, ages 18-25, but did not have active depression at age 26 (past group); and (3) individuals not meeting diagnostic criteria for depression during the transition period. Results highlight serious deficits in psychosocial functioning at age 26 linked to both active and past depression. Although participants with active depression experienced the greatest number of problems, those with past depression evidenced similar deficits across many important domains of functioning. The significant impairments characterizing those with past depression indicate the need for continued monitoring to decrease the risk of recurrence and the establishment of a chronic course of illness. PMID- 16699380 TI - The naturalistic course of unipolar major depression in the absence of somatic therapy. AB - The goal of the study was to describe the naturalistic course of unipolar major depression in subjects not receiving somatic therapy for their depressive illness. Affectively ill individuals were recruited into the Collaborative Depression Study and followed prospectively for up to 15 years. One hundred thirty subjects who recovered from their intake episode of major depression subsequently experienced a recurrence that went untreated for at least 4 weeks following onset of the recurrence. The duration of the recurrent episode was examined using survival analytic techniques. Of the 130 subjects, 46 obtained somatic therapy at some time during the course of their depressive illness, while 84 subjects received no somatic therapy throughout their entire depressive episode. Survival analysis, which accounts for these 46 individuals by censoring their episodes at the time treatment was obtained, yielded a median time to recovery of 23 weeks. In the subsample of 84 subjects whose depressive illness went untreated from its inception through its resolution, the median time to recovery was 13 weeks. These results suggest that there is a high rate of recovery in individuals not receiving somatic treatment of their depressive illness, particularly in the first 3 months of an episode. Because treatment seeking behavior is known to be associated with a worse prognosis, 23 weeks probably represents a lower-limit approximation of the median duration of an untreated depressive episode. PMID- 16699381 TI - Expressed emotion and family interactions in Mexican Americans with schizophrenia. AB - Patients with schizophrenia who are exposed to family environments high in expressed emotion (EE) are at increased risk of relapse. EE is usually measured by an interview with one family member and does not include a direct assessment of family interaction. To understand better the key processes that underlie the relationship between EE and course of illness, we applied a well-validated measure of family interaction, the Structural Family Systems Ratings (SFSR), to 28 Mexican-Americans with schizophrenia and their families. The relationships among EE indices, SFSR components, and relapse were examined. Although measures of EE and family interactions were not related, both EE and one component of the SFSR, family resonance, were associated with a psychotic relapse within 1 year of the assessment. The results suggest that Mexican Americans with schizophrenia benefit from living with families that are able to adapt to the ever-changing circumstances and needs of their ill relatives. PMID- 16699382 TI - Correlates of health-related quality of well-being in older patients with schizophrenia. AB - Research on correlates of health-related quality of life (HRQOL) among older patients with schizophrenia has been very limited. This study evaluated the relative impact of positive, negative, and depressive symptoms, movement disorders, and cognitive impairment on HRQOL among middle-aged and older patients with schizophrenia or schizoaffective disorder. Participants were 199 patients aged 45 to 85 years. The study was cross-sectional. The primary outcome measure was the Quality of Well-Being scale, and correlates were measures of positive and negative symptoms, depression, abnormal movements, and cognitive performance. Severity of depressive symptoms and of cognitive impairment correlated significantly with HRQOL and independently affected HRQOL scores. The initiation/perseveration subscale of the Dementia Rating Scale had the largest impact. These findings suggest that depressive symptoms and cognitive functioning should be part of the routine assessment of older people with schizophrenia and may be targets for psychopharmacological and psychosocial interventions to improve HRQOL. PMID- 16699383 TI - Adult social attachment disturbance is related to childhood maltreatment and current symptoms in borderline personality disorder. AB - We characterized borderline personality disorder (BPD) along two fundamental dimensions of adult social attachment and evaluated attachment associations with childhood maltreatment and current symptoms using self-report measures in 40 outpatients with DSM-IV BPD. The BPD group had significantly greater dimensional attachment impairment and rate of fearful attachment type compared with a healthy control group. Among BPD subjects, dimensional attachment-anxiety was specifically associated with sexual abuse, whereas attachment-avoidance was associated with all five maltreatment types. The two attachment dimensions showed divergent associations with current interpersonal problems, impulsivity subtypes and mood symptoms. We conclude that (1) BPD is characterized by adult attachment disturbance; (2) these attachment problems are strongly related to childhood maltreatment, and to current interpersonal problems and clinical symptoms that are considered core features of BPD; and (3) the diverse problems of BPD patients may arise from two basic mechanisms, each tied to a different type of attachment disturbance, developmental history, and clinical outcome. PMID- 16699384 TI - Childhood physical abuse and differential development of paranormal belief systems. AB - This study compared paranormal belief systems in individuals with and without childhood physical abuse histories. The Revised Paranormal Belief Scale and the Assessing Environments III Questionnaire were completed by 107 University students. Psi, precognition, and spiritualism, which are thought to provide a sense of personal efficacy and control, were among the most strongly held beliefs in abused subjects, and were significantly higher in abused versus nonabused subjects. Superstition and extraordinary life forms, thought to have an inverse or no relation to felt control, were the least strongly held beliefs in abused subjects, and, along with religious beliefs, did not differ between the two abuse groups. Witchcraft was unexpectedly found to be the most strongly held belief among those with abuse histories. Results suggest that by providing a sense of control, certain paranormal beliefs may offer a powerful emotional refuge to individuals who endured the stress of physical abuse in childhood. PMID- 16699385 TI - Depersonalization in psychiatric patients: a transcultural study. AB - There is evidence suggesting that the prevalence of depersonalization in psychiatric patients can vary across cultures. To explore the possible influence of culture on the prevalence of depersonalization, we compared psychiatric inpatient samples from the United Kingdom (N = 31), Spain (N = 68), and Colombia (N = 41) on standardized and validated self-rating measures of dissociation and depersonalization: the Cambridge Depersonalization Scale and the Dissociative Experiences Scale (DES). Colombian patients were found to have lower global scores on the Cambridge Depersonalization Scale and the DES and all its subscales, with the exception of DES-Absorption. No differences were found for measures of depression or anxiety. These findings seem to support the view that depersonalization is susceptible to cultural influences. Attention is drawn to the potential relevance of the sociological dimension "individualism collectivism" on the experience of the self, and it is proposed that cultures characterized by high individualism may confer vulnerability to depersonalization experiences. PMID- 16699386 TI - Dissociation, resting EEG, and subjective sleep experiences in undergraduates. AB - In this study, we explored whether individual differences in dissociation are related to certain resting electroencephalographic (EEG) parameters. Baseline EEG with eyes open and closed was recorded in an undergraduate sample (N = 67). Cortical power in the alpha range was inversely related to dissociative symptoms as measured by the Dissociative Experiences Scale, while both delta and theta power where positively related to dissociation. However, sleep experiences, as indexed with the Iowa Sleep Experiences Survey, were unrelated to resting EEG characteristics. We propose that suppression in the alpha band and raised levels of theta activity, which are typical for high dissociators, might help to explain why dissociative symptoms are accompanied by attentional and memory deficits. PMID- 16699387 TI - Clinical characteristics of perinatal psychiatric patients: a chart review study. AB - Although postpartum depression and other perinatal disorders have been the subject of increased research attention, important questions remain regarding women who actively seek psychiatric treatment during pregnancy and the postpartum period. In this study, we examined clinical records of 500 perinatal psychiatric patients who received treatment in a psychiatric day hospital (N = 398) or outpatient behavioral health clinic (N = 102). Patients' presenting diagnoses, psychiatric history, treatment course, and depressive symptoms were recorded. The majority of women had major depression as their primary diagnosis, with an average Edinburgh Postnatal Depression Scale score of over 20. Many depressed patients were diagnosed with comorbid anxiety and substance abuse disorders. Although most women were willing to take psychotropic medications, a sizable minority were not, particularly those who were breast-feeding. For more than a third of the sample, the treatment sought while pregnant or postpartum represented their first contact with the mental health system. Treatment implications are discussed. PMID- 16699388 TI - A longitudinal perspective on depression and sense of coherence in a sample of mass-evacuated adults from Kosovo. AB - Symptoms of depression and Sense of Coherence (SOC) in a sample of mass-evacuated adults from Kosovo were studied using a prospective design with a baseline study and follow-ups at 3 months and 6 months in Sweden, with an additional follow-up after 1.5 years in both Sweden and Kosovo. The 12-item version of the SOC, depression subscales of the General Health Questionnaire, and the Hopkins Symptom Check List-25 were used. At the additional follow-up after 1.5 years, the same measures were used together with clinical diagnostic assessment. Depression scores increased and SOC decreased over time. There was a significant negative correlation between SOC and symptoms of depression on all occasions, and SOC scores could not predict the diagnosis or symptoms of depression. Those who remained in Sweden became significantly more depressed than those who repatriated. The results are discussed in terms of postmigration stress and breakdown of SOC. PMID- 16699389 TI - Childhood sexual abuse and depressive symptom severity: the role of neuroticism. AB - This study examined neuroticism's role in the association between childhood sexual abuse and severity of depressive symptoms in a sample of 105 psychiatric patients 50 years of age and older diagnosed with major depressive disorder. As hypothesized, men and women who reported histories of childhood sexual abuse had more severe depressive symptoms than those without abuse histories. Further, neuroticism partially accounted for the association between severe childhood sexual abuse and depressive symptom severity. Self-consciousness, a facet of neuroticism conceptually related to shame, also partially accounted for that relationship. These findings suggest that neuroticism may be one way in which childhood sexual abuse contributes to depressive symptoms in later life. Prospective studies are needed. PMID- 16699438 TI - Comparison of multifocal and monovision soft contact lens corrections in patients with low-astigmatic presbyopia. AB - PURPOSE: The purpose of this study was to assess visual performance and patient satisfaction with two presbyopic soft contact lens modalities. METHODS: A crossover study of 38 patients with presbyopia was conducted. Patients were randomized first into either multifocal (Bausch & Lomb SofLens Multifocal) or monovision (SofLens 59) for 1 month. Visual performance was measured with high- and low-contrast visual acuity at distance and near and near stereoacuity. Patients' satisfaction was measured by the National Eye Institute Refractive Error Quality of Life Instrument questionnaire and by recording the patient's final lens preference. RESULTS: Patients maintained at least 20/20 binocular vision with both multifocal (MF) and monovision (MV) contact lenses under high contrast conditions at distance and near. Under low-contrast conditions, patients lost less than a line of vision from the best spectacle correction to either multifocal or monovision contact lens correction at distance (pMF = 0.001, pMV = 0.006). Under low-contrast conditions at near, multifocal wearers lost five to six letters and monovision wearers lost two letters of vision (pMF < 0.001, pMV = 0.03, pMF/MV = 0.005). The average stereoacuity decreased by 79 s arc with monovision vs. multifocal contact lenses (p = 0.002). On the NEI-RQL, patients reported worse clarity of vision (pMF = 0.01, pMV < 0.001), more symptoms (pMF = 0.09, pMV = 0.01), and an improvement in their appearance with contact lens wear (pMF < 0.001, pMV < 0.001). Seventy-six percent of patients reported that they preferred multifocal contact lenses, and 24% preferred monovision contact lenses (p = 0.001). CONCLUSION: The majority of our patients preferred multifocals to monovision, most likely because the Bausch & Lomb SofLens Multifocal provides excellent visual acuity without compromising stereoacuity to the same degree as monovision. PMID- 16699439 TI - The Nepal Longitudinal Study: biometric characteristics of developing eyes. AB - PURPOSE: To identify differences in potential biometric markers for predicting refractive error in school children. METHODS: Biometric data on 895 Tibetan children, aged 6 to 18 years, residing in Katmandu, Nepal, were collected biennially from 1992 to 2000. Measurements included cycloplegic autorefraction, A scan ultrasonography, and video phakometry. Only those children who had been studied at least once at age 12 years or more were included in the analysis. Subjects were divided into two groups: a myopia group if the refractive error was myopic by more than -0.50 D and a nonmyopia group if the refractive error was maximally myopic by -0.50 D, expressed as a spherical equivalent error in the left eye. RESULTS: Biometric measures that differed significantly with increasing age between the two refractive groups included: anterior chamber depth + 0.012 mm/year (p = 0.014), anterior lens radius of curvature + 0.073 mm/year (p = 0.001), lens power -0.059 D/year (p = 0.082), lens thickness -0.005 mm/year (p = 0.02), and vitreous chamber depth + 0.084 mm/year (p < 0.001). Corneal radii of curvature of the myopic group were steeper at all ages by 0.09 mm (p < 0.001), but the rate of change with age was equivalent across the refractive groups. CONCLUSIONS: Compared with those who remained nonmyopic, children who developed myopia had a crystalline lens that was initially thicker and steeper, and a vitreous chamber that was initially shorter. With age, children who became myopic developed greater lens thinning, greater flattening of the anterior lens surface radius, and a greater increase in vitreous chamber depth than their nonmyopic counterparts. PMID- 16699440 TI - Accommodative insufficiency is the primary source of symptoms in children diagnosed with convergence insufficiency. AB - PURPOSE: Accommodative insufficiency (AI) and convergence insufficiency (CI) have been associated with similar symptomology and frequently present at the same time. The severity of symptomology in CI has been linked to the severity of the CI, suggesting a dose-dependent relationship. However, with increasing severity of CI also comes increased comorbidity of AI. AI alone has been shown to cause significant symptomology. We hypothesize that AI drives the symptoms in CI with a comorbid AI condition (CIwAI) and that it is the increased coincidence of AI, rather than increased severity of CI, which causes additional symptomology. METHODS: Elementary school children (n = 299) participated in a vision screening that included tests for CI and AI and the CISS-V15 symptom survey. They were categorized into four groups:1) normal binocular vision (NBV); 2) AI-only; 3) CI only; and 4) CIwAI. One hundred seventy elementary school children fell into the categories of interest. RESULTS: Pairwise comparison of the group means on the symptom survey showed: 1) children with AI-only (mean = 19.7, p = 0.006) and children with CIwAI (mean = 22.8, p = 0.001) had significantly higher symptom scores than children with NBV (mean = 10.3); and 2) children with CI-only (mean = 12.9, p = 0.54) had a similar symptom score to children with NBV. Using a two factor analysis of variance (AI and CI), the AI effect was significant (AI mean = 21.56; no AI mean = 11.56, p < 0.001), whereas neither the CI effect (p = 0.16) nor the CI by AI interaction effect (p = 0.66) were significant. CONCLUSION: CI is a separate and unique clinical condition and can occur without a comorbid AI condition, our CI-only group. Past reports of high symptom scores for children with CI are the result of the presence of AI, a common comorbid condition. When AI is factored out, and children with CI only are evaluated, they are not significantly more symptomatic than children with NBV. PMID- 16699441 TI - Measuring contrast sensitivity under different lighting conditions: comparison of three tests. AB - PURPOSE: The purpose of this study was to evaluate three psychophysical tests for the measurement of contrast sensitivity (CS) and disability glare (DG) at different luminance levels. METHODS: In 60 eyes of 60 individuals (group 1: 20 healthy eyes of young individuals; group 2: 20 healthy eyes of elderly subjects; group 3: 20 eyes with nuclear cataract), CS with best correction was measured twice with the Frankfurt-Freiburg Contrast and Acuity Test System (FF-CATS) and the Functional Acuity Contrast Test (FACT, 1.5 cycles per degree [cpd]) at 167 cd/m2 and 0.167 cd/m2, and with the Pelli-Robson Chart (PRC) at 100 cd/m2 with and without glare. Repeatability of test and retest, and discriminative ability between the different subgroups, were assessed for CS values. RESULTS: Maximum CS values varied across tests. In all groups, highest CS values were obtained with the photopic FF-CATS. For FACT scores at 1.5 cpd, there was a ceiling effect for young subjects. CS scores obtained with the PRC were the lowest. The PRC had the best test-retest repeatability of all tests. Under mesopic conditions with glare, reliability was generally lower; the FF-CATS had the highest repeatability of the mesopic tests. The FF-CATS discriminated best between the different groups for all conditions. CONCLUSIONS: There are large discrepancies in the test results between CS testing methods, especially under different lighting conditions. Results from different CS tests are not interchangeable. PMID- 16699442 TI - The effect of colored lenses on the visual evoked response in children with visual stress. AB - PURPOSE: Some children with visual stress and/or headaches have fewer symptoms when wearing colored lenses. Although subjective reports of improved perception exist, few objective correlates of these effects have been established. METHODS: In a pilot study, 10 children who wore Intuitive Colorimeter lenses, and claimed benefit, and two asymptomatic children were tested. Steady-state potentials were measured in response to low contrast patterns modulating at a frequency of 12 Hz. Four viewing conditions were compared: 1) no lens; 2) Colorimeter lens; 3) lens of complementary color; and 4) spectrally neutral lens with similar photopic transmission. RESULTS: The asymptomatic children showed little or no difference between the lens and no lens conditions. When all the symptomatic children were tested together, a similar result was found. However, when the symptomatic children were divided into two groups depending on their symptoms, an interaction emerged. Children with visual stress but no headaches showed the largest amplitude visual evoked potential response in the no lens condition, whereas those children whose symptoms included severe headaches or migraine showed the largest amplitude visual evoked potential response when wearing their prescribed lens. CONCLUSIONS: The results suggest that it is possible to measure objective correlates of the beneficial subjective perceptual effects of colored lenses, at least in some children who have a history of migraine or severe headaches. PMID- 16699443 TI - Dynamic measurement of accommodation and pupil size using the portable Grand Seiko FR-5000 autorefractor. AB - PURPOSE: The purpose of this study was to evaluate the potential of the portable Grand Seiko FR-5000 autorefractor to allow objective, continuous, open-field measurement of accommodation and pupil size for the investigation of the visual response to real-world environments and changes in the optical components of the eye. METHODS: The FR-5000 projects a pair of infrared horizontal and vertical lines on either side of fixation, analyzing the separation of the bars in the reflected image. The measurement bars were turned on permanently and the video output of the FR-5000 fed into a PC for real-time analysis. The calibration between infrared bar separation and the refractive error was assessed over a range of 10.0 D with a model eye. Tolerance to longitudinal instrument head shift was investigated over a +/-15 mm range and to eye alignment away from the visual axis over eccentricities up to 25.0 degrees . The minimum pupil size for measurement was determined with a model eye. RESULTS: The separation of the measurement bars changed linearly (r2 = 0.99), allowing continuous online analysis of the refractive state at 60 Hz temporal and approximately 0.01 D system resolution with pupils >2 mm. The pupil edge could be analyzed on the diagonal axes at the same rate with a system resolution of approximately 0.05 mm. The measurement of accommodation and pupil size were affected by eccentricity of viewing and instrument focusing inaccuracies. CONCLUSIONS: The small size of the instrument together with its resolution and temporal properties and ability to measure through a 2 mm pupil make it useful for the measurement of dynamic accommodation and pupil responses in confined environments, although good eye alignment is important. PMID- 16699444 TI - Visual acuity in simple myopic astigmatism: influence of cylinder axis. AB - PURPOSE: The relationship between astigmatic refractive errors and their associated visual acuity has been studied in recent years in the context of refractive power space. The influence of the axis of astigmatism remains a matter of controversy. Our goal in this study is to provide additional experimental evidence to clarify this subject. The influence of the simulated axis orientation was compared with other factors that affect visual acuity such as the particular design of the test and the differences between eyes. METHODS: Simple myopic astigmatism from 0 to -3.00 D, in steps of -0.25 D, and with five different axes between 0 degrees and 90 degrees , were simulated on four healthy eyes of young observers. In each case, visual acuity was recorded for three different tests. Refractions were expressed in the form of vectors and visual acuity was represented as a function of strength. RESULTS: No significant differences in visual acuity were found for astigmatism of the same power but different axes. In fact, our results show these differences are even less important than those recorded for the same astigmatism induced in different eyes. The highest discrepancies in visual acuity were found when different charts were used to test the same astigmatic error. CONCLUSIONS: The strength of the vector representing the astigmatic refractive state describes very accurately the performance of visual acuity across simple myopic astigmatic errors. In these cases, visual acuity can be associated with a single refractive parameter. This fact could be useful, especially in statistical studies involving visual performance. PMID- 16699445 TI - Vision loss in younger patients: a review of choroidal neovascularization. AB - Choroidal neovascularization (CNV) is a common cause of vision loss in patients <50 years of age. In these patients, CNV is often the result of pathologic myopia, ocular histoplasmosis syndrome, angioid streaks, or idiopathic causes. Rarely, CNV is seen in children, usually in association with inherited macular dystrophies such as Best's disease (vitelliform macular dystrophy) or drusen of the optic nerve. This article discusses the role of optometrists in the detection and management of CNV. Untreated CNV can cause rapid deterioration of central vision and is associated with a poor prognosis. Optometrists may be able to improve the prognosis of younger patients with CNV by recognizing the signs, symptoms, and risk factors for CNV and promptly referring patients with suspected CNV to a retina specialist. They can also help patients by providing long-term support and rehabilitation. Recent treatment advances, such as the availability of verteporfin (Visudyne; Novartis Pharma AG, Basel, Switzerland) therapy, that are applicable to younger patients with CNV have increased the importance of early detection of CNV and prompt referral of patients to a specialist who can treat CNV. PMID- 16699447 TI - Disassociation between risk of graft loss and risk of non-Hodgkin lymphoma with induction agents in renal transplant recipients. AB - BACKGROUND: It is widely assumed that the graft-enhancing properties of antilymphocyte induction agents and their lymphoma-inducing potential are intimately related. METHODS: The Collaborative Transplant Study (CTS) database was used to evaluate graft survival and non-Hodgkin lymphoma at 3 years according to type of induction in 112,122 patients receiving a deceased-donor renal transplant during 1985 to 2004. RESULTS: The relative risk of 3-year graft loss versus no induction was 1.07 (95% confidence interval [CI], 1.01-1.13; P=0.016) with murine anti-CD3 monoclonal antibody (OKT3), 1.03 (95% CI, 0.95-1.11; NS) with antithymocyte globulin (ATG)-Fresenius, 1.18 (95% CI, 1.02-1.35; P=0.021) with ATGAM, 0.74 (95% CI, 0.68-0.81; P<0.001) with Thymoglobulin, and 0.78 (95% CI, 0.72-0.84; P<0.001) with interleukin (IL)-2RA induction. The standardized incidence ratio of lymphoma compared with a similar nontransplant population was 21.5 (95% CI, 15.7-28.8; P<0.001) with OKT3, 4.9 (95% CI, 1.6-11.5; P=0.008) with ATG-Fresenius, 29.0 (95% CI, 12.5-57.1; P<0.001) with ATGAM, 21.6 (95% CI, 14.3 31.2; P<0.001) with Thymoglobulin, 7.8 (95% CI, 4.4-12.9; P<0.001) with IL-2RAs, and 9.4 (95% CI, 8.3-10.6 P<0.001) with no induction. CONCLUSIONS: Those agents that offered the highest rates of graft survival were not necessarily associated with the highest risk of lymphoma. Graft survival was significantly improved with Thymoglobulin and IL-2RA induction, whereas lymphoma rates were highest with ATGAM, OKT3, and Thymoglobulin. IL-2RA agents seem to offer the best risk-to benefit ratio for this patient population overall in terms of graft survival and lymphoma. PMID- 16699448 TI - Target of rapamycin inhibitors (sirolimus and everolimus) for primary immunosuppression of kidney transplant recipients: a systematic review and meta analysis of randomized trials. AB - BACKGROUND: Target of rapamycin inhibitors (TOR-I) have a novel mode of action but uncertain clinical role. We performed a systematic review of randomized trials where immunosuppressive regimens containing TOR-I were compared with other regimens as initial therapy for kidney transplant recipients. METHODS: Databases (inception, June 2005) and conference proceedings (1996-2005) were searched. Two independent reviewers assessed trials for eligibility and quality. Results at 1 year, are expressed as relative risk (RR), where values<1 favor TOR-I, or lower dose of TOR-I, and for continuous outcomes are expressed as weighted mean difference (WMD), both expressed with 95% confidence intervals (CI). RESULTS: Thirty-three trials (142 reports) were included (27 trials of sirolimus, 5 of everolimus, and 1 of head-to-head comparison). When TOR-I replaced calcineurin inhibitors (CNI) (8 trials with 750 participants), there was no difference in acute rejection (RR, 1.03; 95% CI, 0.74-1.44), but serum creatinine was lower (WMD, -18.31 micromol/L; 95% CI, -30.96 to -5.67) and bone marrow more suppressed (leukopenia: RR 2.02; 95% CI, 1.12-3.66; thrombocytopenia: RR, 6.97; 95% CI, 2.97 16.36; and anaemia: RR, 1.67; 95% CI, 1.27-2.20). When TOR-I replaced antimetabolites (11 trials with 3966 participants), acute rejection and cytomegalovirus infection (CMV) were reduced (RR, 0.84; 95% CI, 0.71-0.99; RR, 0.49; 95% CI, 0.37-0.65, respectively), but hypercholesterolemia was increased (RR, 1.65; 95% CI, 1.32-2.06). When low- was compared with high-dose TOR-I, with equal CNI dose (10 trials with 3,175 participants), rejection was increased (RR, 1.23; 95% CI, 1.06-1.43) but calculated glomerular filtration rate (GFR) higher (WMD, 4.27 mL/min; 95% CI, 1.12-7.41), and when lower-dose TOR-I and standard dose CNI were compared with higher-dose TOR-I and reduced CNI, acute rejection was reduced (RR, 0.67; 95% CI, 0.52-0.88), but calculated GFR was also reduced (WMD, -9.46 mL/min; 95% CI, -12.16 to -6.76). There was no significant difference in mortality, graft loss, or malignancy risk for TOR-I in any comparison. CONCLUSIONS: TOR-I have been evaluated in four different primary immunosuppressive algorithms: as replacement for CNI and antimetabolites, in combination with CNI at low and high doses, and with a variable dose of CNI. Generally, surrogate endpoints for graft survival favor TOR-I (lower risk of acute rejection and higher GFR), and surrogate endpoints for patient outcomes are worsened by TOR-I (bone marrow suppression and lipid disturbance). Long-term hard endpoint data from methodologically robust randomized trials are still required. PMID- 16699449 TI - Attitudes toward financial incentives, donor authorization, and presumed consent among next-of-kin who consented vs. refused organ donation. AB - BACKGROUND: Financial incentives, donor authorization, and presumed consent are strategies designed to increase organ donation rates. Surveys designed to assess attitudes toward these initiatives have been conducted with the general public, transplant patients, and transplant professionals. METHODS: To assess attitudes toward financial incentives, donor authorization, and presumed consent and to identify multivariate predictors of such attitudes, we conducted telephone interviews with 561 family members who had recently been asked for consent to donate the organs of a deceased family member (348 donors, 213 nondonors). RESULTS: Financial incentives would have made a difference in the donation decision for 54% of nondonors (vs. 46% of donors, P=0.02), and a higher percentage of nondonors would themselves become donors if financial incentives were available (P=0.03). Donors had significantly more favorable attitudes toward donor authorization (P<0.0001) and presumed consent (P<0.0001) policies. Overall, 54% of participants thought that family permission for donation was unnecessary when the deceased documented their donation intention, and 24% favored a presumed consent law with an opting out provision. CONCLUSIONS: Of the three initiatives, donor authorization is likely supported by more donor and nondonor families than either financial incentives or presumed consent. Public education efforts should aim to better inform the public regarding existing and proposed donor authorization legislation and its benefits for registered organ donors and their families. PMID- 16699450 TI - TNFalpha and IL-10 in bone marrow transplantation: does genotype matter? PMID- 16699451 TI - Kidney donation--all gifts have a price. PMID- 16699452 TI - Association of TNFd and IL-10 polymorphisms with mortality in unrelated hematopoietic stem cell transplantation. AB - BACKGROUND: Non-HLA immunogenetic polymorphisms may influence outcome of hematopoietic stem cell transplantation (HSCT). In this study, we have determined the role of TNFa, TNFd, IL-10, IL-1, IL-1Ra, and IL-4R polymorphisms in patients transplanted with HSC of an unrelated donor. METHODS: The allelic variants of four SNPs (IL-10-1082, IL-1beta-511, IL-4R-3223, IL-4R-1902) and four microsatellites (TNFa, TNFd, IL-10-1064, IL-1Ra) were determined in 131 unrelated patient/donor pairs typed for HLA-A/B/C/DR/DQ (four digits). RESULTS: The allelic distribution of the polymorphisms was similar to that previously reported in Caucasoid populations. Patient and donor TNFd and patient IL-10-1064 polymorphisms correlated with mortality in univariate analysis. Patients with TNFd1/d2/d3 genotypes had 3-year survival rates of 65%. A gradual decrease in survival rates was observed for patients with TNFd3/d3 genotypes (50%, p=n.s.), TNFd4 (46%, P=0.08), and TNFd5 (33%, P=0.03). A multivariate analysis of 10/10 matched patients revealed that the following patient genotypes correlated with lower survival: TNFd3/d3 (RR 4.08, P=0.026) TNFd4 (RR 3.78, P=0.032) and TNFd5 (RR 6.69, P=0.021) all compared to TNFd1/d2/d3 genotypes. Patient IL-10 (12, 14, 15) microsatellite alleles correlated with lower 3-year survival (28%) when compared to IL-10 (<12) (56%, P=0.052) and to Il-10 (13) alleles (60%, P=0.0023). In multivariate analysis this correlation remained significant only in recipients of HSCT of 10/10 HLA matched donors (RR=2.96, P=0.038). CONCLUSION: The data demonstrate a significant correlation of the TNFd and IL-10-1064 microsatellite polymorphisms with mortality after unrelated HSCT. They support the hypothesis that simple genomic tests, in addition to precise HLA matching, may contribute to determine prognosis in patients undergoing unrelated HSCT. PMID- 16699453 TI - The impact of living-related kidney transplantation on the donor's life. AB - BACKGROUND: Living-donation kidney transplantation (LDKT) is increasingly performed for treatment of chronic renal failure. Recently, risks for the donor and problems in decision-making have been stressed. This study was conducted to illuminate the decision making-process and consequences of LDKT on family life, the financial and occupational situation. Moreover, quality of life (QOL) and mental distress were explored. METHODS: All German residents at Essen University, who donated their kidney between 1999 and 2003, were included in the study. Donors filled out the questionnaire of the European Multicenter Study of Transplantation Using Living Donors, the Short Form 36-Health Survey, and the Brief Symptom Inventory. RESULTS: Out of a total of 65 donors, 47 replied (72%) at an average 2.5 years postdonation. No fatalities occurred in the whole sample (n=65), medical complications were experienced by 28%. Most donors decided voluntarily (94%) and spontaneously (66%) to donate, after donation 96% stated that they would decide in the same way again. QOL was within the norm. On the other hand, 10% experienced family conflicts, every eighth donor suffered from clinically relevant distress, financial disadvantages were experienced by every fourth donor, with 25% not answering this question. CONCLUSION: Seen from the donor's perspective, LDKT is a relatively safe procedure. However, increased rates of donors with mental distress and intra-familial conflicts emphasize the need for a careful selection process. Regular postdonation psychosocial screening and provision of specific interventions to those in need are recommended. Donors should not suffer from financial and occupational disadvantages resulting from donation. PMID- 16699454 TI - Early increase of retinal arterial and venous blood flow velocities at color Doppler imaging in brittle type 1 diabetes after islet transplant alone. AB - Little information is currently available about the role of islet transplantation alone (ITA) on the retinal microcirculation. Our purpose was to investigate with color-Doppler-imaging the effect of ITA after one year on the blood flow velocities of central retinal artery and vein. Central retinal arteries and veins of both eyes of 10 ITA patients were evaluated with color-Doppler-imaging before and one year after transplant. Peak systolic velocity (psv), end diastolic velocity (edv) for arteries and maximum velocity (maxv), minimum velocity (minv) for veins were recorded and compared with a control group of type 1 diabetic patients. At one year, a statistically significant increase of blood flow velocities of central retinal arteries (psv: 6.09+/-0.46 vs. 10.12+/-1.20 cm/s, P=0.01) and veins (maxv: 3.12+/-0.28 vs. 6.12+/-1.00 cm/s, P=0.01) was found only in the ITA patients. An early, significant increase of arterial and venous retinal blood flow velocities was found after ITA. PMID- 16699455 TI - The routine use of high-resolution immunological screening of recipients of primary deceased donor kidney allografts is cost-effective. AB - BACKGROUND: The economic and health benefits of kidney transplantation are dependent on the length of allograft survival. High-resolution immunological screening can identify recipients at increased risk of early graft loss caused by acute rejection, but the use of these tests increases screening costs before transplantation. The objective of this study was to evaluate the cost effectiveness of routine use of high-resolution flow-cytometry cross-matching and solid-phase screening for all recipients of primary deceased donor kidney transplants. METHODS: A Markov model was constructed to evaluate costs and effects of two different clinical strategies on a simulated cohort of 1,000 transplant recipients: serological screening (SS) only and flow screening (FS) only. Outcomes measures were total cost of patient care over 25 years, life expectancy, quality-adjusted life expectancy, and transplant life expectancy. RESULTS: In the base-case analysis, FS was associated with an average gain of 0.08 life years, 0.25 transplant life years, and 0.08 quality-adjusted life years per patient. SS was associated with a higher cost of CND$6,397 per patient, mostly because of increased use of dialysis in patients who suffered early graft loss under the SS strategy. The results were robust to uncertainty in the majority of variables, and a strategy using FS was cost-effective except under the unlikely scenario where the false-negative rate for SS was or=60 years). Independent variables included social support (measured with the Social Support List Discrepancies questionnaire), sociodemographic and medical variables, side effects and compliance. RESULTS: Predictors of better PHS in patients<40 years were better social support (P 70% of their person-years since 1960 (181 cases, 142 controls). For those in the diet arm of the study (458 cases, 383 controls from 4 centers) and for Iowa participants in both the diet and drinking water analyses, we estimated dietary nitrate and nitrite intake using a 117-item food-frequency questionnaire that included foods high in nitrate and nitrite. Odds ratios and 95% confidence intervals were calculated using logistic regression, adjusting for the study matching factors, education, and caloric intake (diet analyses only). RESULTS: We found no overall association with the highest quartile of average drinking water nitrate (> 2.90 mg/L nitrate-N: odds ratios = 1.2; 95% confidence interval = 0.6-2.2) or with years > or = 5 mg/L (10+ years: 1.4; 0.7-2.9). We observed no evidence of an interaction between drinking water nitrate exposure and either vitamin C or red meat intake, an inhibitor and precursor, respectively, of N-nitroso compound formation. Among those in the diet arm, dietary nitrate was inversely associated with risk of NHL (highest quartile: 0.54; 0.34-0.86). Dietary nitrite intake was associated with increasing risk (highest quartile: 3.1; 1.7-5.5) largely due to intakes of bread and cereal sources of nitrite. CONCLUSION: Average drinking water nitrate levels below 3 mg/L were not associated with NHL risk. Our study had limited power to evaluate higher levels that deserve further study. PMID- 16699474 TI - Liver disease: hepatitis B. Treatment of chronic hepatitis B With entecavir. PMID- 16699475 TI - Osteoporosis in patients with inflammatory bowel disease: risk factors, prevention, and treatment. AB - Patients with inflammatory bowel disease (IBD) are at increased risk for osteoporotic fracture. Bone density testing and osteoporosis management are recommended for IBD patients at greater risk for fracture (ie, postmenopausal women, men aged . 60 years, and those with low body mass indices, glucocorticoid use, family history of osteoporosis, and malabsorption). Patient management includes modification of osteoporosis risk factors, such as calcium and vitamin D supplementation, hormone deficiency correction, and smoking cessation. When indicated, bisphosphonates, such as risedronate and alendronate, have been shown to increase bone mass and reduce fracture risk in patients with glucocorticoid induced osteoporosis. Infliximab, an anti-tumor necrosis factor a antibody, increases bone mineral density, but this effect has not as yet translated into reduced fracture risk. PMID- 16699476 TI - A unifying hypothesis for the functional gastrointestinal disorders: really multiple diseases or one irritable gut? AB - The functional gastrointestinal disorders are defined by the Rome criteria as a heterogeneous group of symptom-based conditions that have no structural or biochemical explanation. However, this definition now seems outdated, because structural and molecular abnormalities have begun to be recognized in subsets of patients with the irritable bowel syndrome (IBS), the prototypic functional bowel disease. A complex classification system based arbitrarily on symptom criteria does not fit in with a number of emerging facts. For example, the symptom overlap of IBS with gastroesophageal reflux disease is not due to chance, and the emergence of post-infectious IBS, dyspepsia, or both after Salmonella gastroenteritis fits better with a 1-disease model. A new paradigm seems to be needed. All of these disorders may arise after infection or gut inflammation, but the phenotype depends on localized neuromuscular dysfunction in the predisposed human host (the "irritable gut"). PMID- 16699477 TI - Clostridium difficile-associated diarrhea: resurgence with a vengeance. AB - There has been a significant increase in the incidence and severity of Clostridium difficile-associated diarrhea (CDAD) in the past several years, including outbreaks in multiple states and provinces in the United States and Canada, as well as in the United Kingdom. A new, highly virulent strain of C. difficile has appeared that is less responsive to standard therapy and associated with a high rate of recurrence. Along with nosocomially acquired infections there has been a rise in the number of community-acquired cases of CDAD, even among those without prior antibiotic exposure. Many factors have contributed to this epidemic, including the development of resistance to the widely used fluoroquinolones class of antibiotics. Because this new strain is less responsive to standard therapy, particularly metronidazole, a number of new antibiotics and other therapies are actively being investigated for use in both primary and recurrent CDAD. A multifaceted approach to managing CDAD is called for, including active surveillance, antibiotic stewardship, and meticulous attention to contact precautions, including gloves, gowns, and hand washing. PMID- 16699479 TI - Pancreatobiliary disease. Metallic stents in the therapy of benign biliary strictures: a new twist. PMID- 16699478 TI - Treatment of ulcerative colitis with oral mesalamine: advances in drug formulation, efficacy expectations and dose response, compliance, and chemoprevention. AB - Sulfasalazine, olsalazine, balsalazide, delayed-release mesalamine, controlled release mesalamine, mesalamine pellets, and Multi-Matrix System mesalamine are effective first-line therapies for the treatment of mildly to moderately active ulcerative colitis and for subsequent maintenance of remission. For induction therapy it is unclear if there is a dose response above 1.5 g, and for maintenance therapy existing data do not support a dose response above 1.5 g. Sulfasalazine has more frequent side effects than olsalazine, balsalazide, and mesalamine formulations. Once-daily dosing with multi-matrix system mesalamine 1.2 g tablets may lead to optimal compliance. Mesalamine >/= 1.2 g and sulfasalazine >/= 2 g reduce the risk of colorectal cancer in patients with ulcerative colitis. Drug formulations, efficacy expectations and dose response, toxicity expectations, compliance considerations, and chemoprevention considerations are reviewed. PMID- 16699481 TI - Infectious disease. Failures of antibiotic prophylaxis for travelers' diarrhea: norovirus may be the culprit. PMID- 16699482 TI - [International rule on access and benefit sharing under the Convention on Biological Diversity]. PMID- 16699483 TI - [Survey of Trichophyton tonsurans infection in Japan. Molecular epidemiology and factors affecting adequate hairbrush sampling]. AB - At the 48th Annual Meeting of The Society for Japanese Medical Mycology, held in October, 2004, we reported our findings from a survey on Trichophyton tonsurans infections in the Hokuriku and Kinki regions of Japan. The survey revealed that a few epidemics had occurred across these regions. In this article, we introduce our subsequent studies relating to 1) molecular epidemiology of isolates taken from people in many parts of Japan and 2) factors affecting adequate sampling of the scalp with hairbrushes, essential for surveying and monitoring the infection. In total, 198 isolates of Trichophyton tonsurans were analyzed using restriction fragment length polymorphisms of the non-transcribed spacer regions of ribosomal RNA genes. The restriction enzyme Mva I indicated two molecular types of strains, implying that the causative agents of the epidemic had different origins. None of the isolates obtained from the epidemic showed the same restriction profile as that of isolates from aged and sporadic cases. The published hairbrush method suitable for obtaining samples from the scalp of Judo trainees was reevaluated by changing several factors. We found that sampling should not be done soon after the students' physical training because other fungal elements may give a false positive, samples should not be obtained from students who have recently applied topical antimycotics, and samples should be taken under the guidance of qualified instructors familiar with the sampling method. PMID- 16699484 TI - [Dermatomycosis and environment]. AB - Environments may act as reservoirs for pathogenic fungi, a determinant of the establishment of fungal infection, or an exacerbating factor of disease. In recent years, skin disease caused by geophilic fungi has been decreasing, while case reports of zoonoses from various animals are increasing. Outbreaks of anthropophilic T. tonsurans infection pose a problem to medical mycologists. Tinea pedis is the most common exogenous dermatomycosis in Japan. Although T. rubrum is presumed to be the dominant pathogen of this disease, T. mentagrophytes is detected more frequently from various environments, so far, the reason for this discrepancy has not been fully understood. The latest knowledge about the route of dermatophyte foot infection is as follows: (1) Dermatophyte propagules disseminated from patients may contaminate not only bath-mats but also wood floors, Japanese style mattings, concrete floors, slippers, cushions, etc., and from them adhere to healthy skin. (2) The agar stamping method can easily detect dermatophytes from the skin and the environment. (3) Propagules of T. mentagrophytes can survive for more than three months under certain conditions such as in rubber boots. (4) In order to eliminate dermatophytes gathered in socks and footwear, simple procedures (washing, bathing with hot water, or wiping with a towel) are all effective. (5) Prior application of an antifungal agent promptly eradicates dermatophyte propagules adhering to the skin from the environment. The author also mentioned the possibility of asymptomatic dermatophyte colonization, and the high prevalence of dysgeucia in oral carriers of Candida albicans. PMID- 16699485 TI - [Onychomycosis: recent progress in the epidemiology, diagnosis and treatment]. AB - Recent progress in the epidemiology, diagnosis and treatment of onychomycosis is summarized. The risk factors of this condition were investigated, and the results of analyses of the systemic state of patients, shape of nails, blood circulation around nails, nail growth rate and causative fungi were reported. In making a diagnosis of onychomycosis, the major effect of the quality of collected nail samples on the results was reconfirmed. In addition to the KOH method and culture method, a molecular biological method was introduced. From the therapeutic perspective, 1) prevention of recurrence, 2) identification of patients with high risk of onychomycosis, and 3) attempting new treatments (development of new drugs, improvement of administration and dosage, and evaluation of combined therapy) were discussed. The effectiveness of concomitant use with nail lacquer is anticipated in combined therapy. New studies on accurate diagnosis, early detection and early stage treatment are now being undertaken. PMID- 16699486 TI - [Malassezia related diseases]. AB - The genus Malassezia is now divided into eleven species. Different species initiate or aggravate different skin diseases. In seborroheic dermatitis, M. restricta play an important role, while in atopic dermatitis, M. globosa and/or M. restricta are major cutaneous microflora. M. globosa is a causative species of tinea versicolor, and this species is also a causative species of malassezia folliculitis. We should therefore obtain better knowledge of the ecological and pathogenic roles of malassezia. PMID- 16699487 TI - [Dermatological zoonosis by fungi in Japan]. AB - Dermatophytosis, cryptococcosis and sporotrichosis are dermatological zoonosis caused by fungi. Control of dermatophytosis is still difficult, although it is well known as a transmittable disease from animal to human. Exotic animals such as hamster, guinea pig and rabbit, some imported from foreign countries, have become a frequent source of human dermatophytosis, and the emergence of fungal pathogens followed. The nasal cavity of dogs and cats is an important reservoir of pathogens of cryptococcosis, in addition to bird droppings. Though there have been many reports of human sporotrichosis caused by patients' pet, especially cats in Europe and U.S., since there has been no report of zoonotic transmission of sporotrichosis, zoonosis is not believed do be present in Japan. The interrelation and cooperation of medical and veterinary doctors and all other scientists involved is therefore required to prevent fungal zoonosis, because the condition may be partly due to changes in the environments of human and animal life and the more intimate relationships between humans and animals. PMID- 16699488 TI - First clinical isolates of Nocardia carnea, Nocardia elegans, Nocardia paucivorans, Nocardia puris and Nocardia takedensis in Japan. AB - Five aerobic actinomycete strains isolated from patients in Japan were assigned provisionally to the genus Nocardia based on morphological and physiological characteristics. The five strains, IFM 10481, IFM 0668, IFM 0901, IFM 0583 and IFM 0342, were not classified into any Nocardia species reported as infectious agents in Japan. Therefore, they were studied further to determine their specific taxonomic positions. Detailed chemotaxonomic and physiologic characterization and 16S rDNA sequence data of the five strains showed that they belonged to respective species of Nocardia carnea, N. elegans, N. paucivorans, N. puris and N. takedensis. This is the first isolation report of these five Nocardia species from patients in Japan. PMID- 16699489 TI - A novel method to estimate the contribution of the vapor activity of essential oils in agar diffusion assay. AB - By the combined use of agar diffusion, agar vapor and agar vapor-inhibitory assays, contribution of the vapor activity of essential oils was quantitatively estimated. The test organisms were Trichophyton mentagrophytes and Aspergillus fumigatus. Agar vapor assay was used to confirm the vapor activity of the oils. The parameter delta defined as a contribution index of the vapor activity was calculated by (1 - b-c/a-c) x 100, where a is inhibitory diameter in the diffusion assay, b is inhibitory diameter in the vapor-inhibitory assay and c is diameter of the sealed ring in the vapor-inhibitory assay (21 mm). Many of the essential oils examined showed a delta value near 100, thus providing the major contribution of the vapor activity to the inhibitory diameter. Essential oils containing aldehyde as major constituent showed low delta value, indicating the major inhibition was due to agar diffusion. Major essential oil components behaved similarly; the delta value was increased in the following order: aldehyde < phenol < alcohol < ester, oxide, hydrocarbon, indicating the enhanced contribution of the vapor activity in that order. PMID- 16699490 TI - [A case of onychomycosis induced by Scopulariopsis brevicaulis]. AB - The patient was a 57-year-old woman who initially consulted our department on May 31, 2002 with a chief complaint of other dermatoses. The initial examination demonstrated deformation of approximately 1/3 of the inner part of the nail plate in the right hallux, in addition to brown discoloration and keratin hypertrophy under the deformed nail plate. KOH-prepared direct microscopy revealed the presence of large round spores and hyphae. Since colonies similar to those of Trichophyton rubrum were obtained by culture, daily administration of terbinafine (125 mg/day) was initiated from June 17, 2002 based on a suspicion of tinea unguium induced by T. rubrum. However, the morphology of the colonies began to change toward a brown powder-like configuration after 2-3 weeks of culture, and band forms were also detected by slide culture, resulting in the diagnosis of onychomycosis induced by Scopulariopsis brevicaulis. From August 6, 2002, the dose of terbinafine was increased to 250 mg/day, which was maintained until November 22, 2002. When the patient visited our department approximately 2 months later, KOH-prepared direct microscopy revealed that she was still positive for S. brevicaulis. Therefore, terbinafine (250 mg/day) was administered again for 1 month from April 25, 2003, followed by a judgment of complete healing on July 4, 2003. Thereafter, there was no recurrence of onychomycosis up until January 29, 2005. PMID- 16699491 TI - [An epidemiological survey of dermatomycoses in Japan, 2002]. AB - An epidemiological survey of dermatomycoses and the causative fungus flora of dermatophytoses in Japan for 2002 was made on a total number of 72,660 outpatients who visited 14 dermatological clinics throughout Japan. The results were as follows: 1) Dermatophytosis was the most prevalent cutaneous fungal infection (7,994 cases) seen in these clinics, followed by candidiasis (755 cases) and then Malassezia infections (220 cases). 2) Among dermatophytoses, tinea pedis was the most frequent (4,813 cases: male 2,439, female 2,374), then in decreasing order, tinea unguium (2,123 cases: male 1,093, female 1,030), tinea corporis (497 cases: male 281, female 216), tinea cruris (299 cases: male 249, female 50), tinea manuum (248 cases: male 144, female 104) and tinea capitis including kerion (14 cases, male 6, female 8). 3) Tinea pedis and tinea unguium are seen to increase in summer season, among the aged population and among males in each clinic. When compared to the previous surveys (1992 and 1997) by clinical form, t. unguium patients increased from 1.9% of total outpatients in 1992, to 2.0% in 1997, then to 2.9% in 2002. 4) As the causative dermatophyte species, Trichophyton rubrum was the most frequently isolated among all dermatophyte infections except tinea capitis. 5) T. rubrum was isolated from 63.3%(1,431/2,262) of tinea pedis lesions, followed by Trichophyton mentagrophytes (36.6%, 829/2,262), and also 88.8% (325/366) of t. corporis, 95.4% (185/194) of t. cruris and 85.6% (462/540) of t. unguium. 6) Cutaneous candidiasis was seen in 755 cases (1.0%) of 72,660 outpatients. Intertrigo (347 cases) was the most frequent clinical form, followed by erosio interdigitalis (103 cases) and diaper candidiasis (102 cases). It has a tendency to affect the aged being complicated with topical predisposing factors. 7) Cutaneous Malassezia infections and other superficial fungal infections are seen in 220 cases, without any characteristic features by gender or clinical form. PMID- 16699492 TI - Reexamination of Coccidioides spp. reserved in the Research Center for Pathogenic Fungi and Microbial Toxicoses, Chiba University, based on a multiple gene analysis. AB - The Research Center for Pathogenic Fungi and Microbial Toxicoses, Chiba University is the only organization in Japan to possess a series isolates of Coccidioides spp., which are the most virulent pathogenic fungi and which are treated as biosafety level 3 microorganisms. Recently, the genus Coccidioides has been classified into two species, C. immitis and C. posadasii, based on their endemic areas and genotyping; the former species is endemic to the state of California, and the latter is endemic to other parts of North and South America. We reevaluated 19 isolates of Coccidioides immitis stored in our center using a multiple gene analysis. Five isolates were identified as C. immitis and 14 as C. posadasii. Their sequence information in GenBank will help to identify the two genospecies of Coccidioides spp. PMID- 16699493 TI - Application of intravascular ultrasound in anti-atherosclerotic drug development. AB - The background use of a number of established therapies presents a key challenge for the development of novel anti-atherosclerotic agents: how to predict potential efficacy before the completion of long-term trials with endpoints such as mortality. This challenge has stimulated the search to develop intermediate measures of efficacy. Recent advances now allow intravascular ultrasound (IVUS) to provide an accurate assessment of atheroma accumulation within the arterial wall. Here we describe how IVUS can be applied to the serial assessment of atheroma burden in response to treatment with a range of anti-atherosclerotic strategies, which has resulted in its emergence as a key technology in the evaluation and approval of novel drugs. PMID- 16699494 TI - Impact of genetic diagnostics on drug development strategy. AB - Innovation in diagnostics will be essential for the successful adoption of personalized medicine and will also have a major impact on the success of drug development strategies. To remain competitive, companies will need to embrace the rapid innovation in the diagnostic market, utilize diagnostic tools concurrently with research and development, and re-invent how new drugs are brought to market. In this article, specific examples of targeted oncology drugs are used to illustrate the general impact of genetic diagnostics on drug development. PMID- 16699495 TI - An adhesion molecule in free-living Dictyostelium amoebae with integrin beta features. AB - The study of free-living amoebae has proven valuable to explain the molecular mechanisms controlling phagocytosis, cell adhesion and motility. In this study, we identified a new adhesion molecule in Dictyostelium amoebae. The SibA (Similar to Integrin Beta) protein is a type I transmembrane protein, and its cytosolic, transmembrane and extracellular domains contain features also found in integrin beta chains. In addition, the conserved cytosolic domain of SibA interacts with talin, a well-characterized partner of mammalian integrins. Finally, genetic inactivation of SIBA affects adhesion to phagocytic particles, as well as cell adhesion and spreading on its substrate. It does not visibly alter the organization of the actin cytoskeleton, cellular migration or multicellular development. Our results indicate that the SibA protein is a Dictyostelium cell adhesion molecule presenting structural and functional similarities to metazoan integrin beta chains. This study sheds light on the molecular mechanisms controlling cell adhesion and their establishment during evolution. PMID- 16699497 TI - Epigenetic profiling of synchronous colorectal neoplasias by quantitative DNA methylation analysis. AB - Transcriptional inactivation of tumor-suppressor genes by promoter CpG island methylation is thought to be an important mechanism in human carcinogenesis. The CpG island methylator phenotype (CIMP) with extensive promoter methylation appears to be a distinct epigenetic subtype of colorectal carcinoma. Most previous studies on CpG island methylation in colorectal carcinoma used methylation-specific PCR, which may detect low levels of DNA methylation with little or no biological significance. In contrast, quantitative DNA methylation assays have been shown to provide useful information beyond that which can be achieved with methylation-specific PCR. Synchronous neoplasias provide a unique model for investigators to examine molecular alterations in multistep tumorigenesis within one individual. However, no study to date has quantified DNA methylation of CIMP-specific promoters in synchronous colorectal neoplasias. Utilizing real-time PCR (MethyLight), we quantified DNA methylation in five CIMP specific gene promoters [CACNA1G (calcium channel, voltage-dependent, T type alpha-1G subunit), CDKN2A (p16/INK4A), CRABP1 (cellular retinoic acid binding protein-1), MLH1 and NEUROG1 (neurogenin 1)] and MGMT in six synchronous carcinoma pairs (12 carcinomas) and eight synchronous carcinoma and adenoma pairs (16 tumors). We found that while some synchronous tumor pairs showed discordant promoter methylation patterns, other tumor pairs showed similar, but not exactly identical, patterns of promoter methylation. All but two pairs showed concordant patterns of CIMP status (CIMP positive vs CIMP negative) (P = 0.05 in cancer pairs). BRAF mutations were present in only CIMP-positive tumors. A high degree of microsatellite instability (MSI-H) was observed in both CIMP-positive and CIMP negative tumors. KRAS mutations were not concordant in any synchronous neoplasia pair. In conclusion, epigenetic alterations at CIMP-specific promoter CpG islands in synchronous colorectal neoplasias likely have both random and nonrandom components. PMID- 16699496 TI - Transformation-specific matrix metalloproteinases, MMP-7 and MMP-13, are present in epithelial cells of keratoacanthomas. AB - Keratoacanthomas are rapidly growing hyperproliferative skin tumors that may clinically or histologically be difficult to distinguish from well-differentiated squamous cell cancers (SCCs). UV light, trauma, and immune suppression represent their etiological factors. As matrix metalloproteinases (MMPs) are implicated at all stages of tumorigenesis, we investigated the expression profile of several cancer-related MMPs to find markers that would differentiate keratoacanthomas from SCCs and shed light to the pathobiology of keratoacanthoma. Samples from 31 keratoacanthomas and 15 grade I SCCs were studied using immunohistochemistry for MMP-2, -7, -8, -9, -10, -13, and -19 and p16 and laminin-5gamma2 chain. In situ hybridization for MMP-7, -10, and -13 was performed in a subset of tumors. Keratinocytes with atypia, presence of neovascularization, and composition of the inflammatory infiltrate were graded from hematoxylin-eosin stainings. MMP-7 was present in the epithelium of 4/31 keratoacanthomas and 9/15 SCCs, MMP-8 in 3/30 keratoacanthomas and 0/15 SCCs, but MMP-13 in 16/31 keratoacanthomas and 10/15 SCCs, and MMP-10 in 28/31 keratoacanthomas and all cancers. MMP-9 was detected in the epithelium in 5/31 keratoacanthomas and 8/15 SCCs, whereas MMP-2 was only present in fibroblasts in both tumors. MMP-19 was upregulated in proliferating epithelium of keratoacanthomas as was p16. Cytoplasmic laminin-5gamma2 was particularly abundant in keratinocytes at the pushing border of MMP-13-positive keratoacanthomas. We conclude that although some MMPs (MMP-10 and -13) are abundantly expressed in keratoacanthomas, the presence of MMP-7 and -9 in their epithelial pushing border is rare and should raise suspicion of SCC. Further, the loss of MMP-19 and p16 could aid in making the differential diagnosis between well-differentiated SCC and keratoacanthoma. Frequent expression of the transformation-specific MMP-13 in keratoacanthomas suggests that they are not benign tumors but incomplete SCCs. PMID- 16699498 TI - Effects of interferon plus ribavirin treatment on NF-kappaB, TGF-beta1, and metalloproteinase activity in chronic hepatitis C. AB - Little is known about the cellular and molecular mechanisms underlying the effects of anti-viral therapy on the regression of liver inflammation and fibrosis in chronic hepatitis C. The aim of this study was to evaluate the effects of interferon alpha and ribavirin in combination therapy on the tissue expression of nuclear-factor kB (NF-kappaB) (a transcription factor coordinating the expression of stress genes involved in immune response and inflammation), of the polypeptide transforming growth factor beta-1 (TGF-beta1) and matrix metalloproteinases 1 (MMP-1) (both of which play an important part in the pathological process of liver fibrogenesis), and on the serum levels of soluble TGF-beta1, tissue inhibitors of metalloproteinases (TIMP)-1, and active endogenous MMP-2 and MMP-9 in paired (pre- and post-treatment) liver biopsy and serum samples of subjects with chronic hepatitis C. Serum levels of TGF-beta1, TIMP-1, MMP-2, and MMP-9 were evaluated by enzyme-linked immunosorbent assay. Liver expression of muscle-specific alpha-actin, NF-kappaB, TGF-beta1, and MMP-1 was studied immunohistochemically using commercially available mono- and polyclonal antisera in an avidin-biotin complex method. Combination therapy induced a reduction in the liver expression of TGF-beta and NF-kappaB and an increased expression of MMP-1, regardless of the virological response to the treatment. The greater expression of MMP-1 and lesser expression of NF-kappaB were both associated with an improvement in fibrosis score. These effects paralleled the significant increase in soluble MMP-9/TIMP-1 ratio in post-therapy sera. Combination therapy with interferon and ribavirin affects the tissue expression of TGF-beta-1 and NF-kappaB and favors metalloproteinase activity, and may thereby modulate hepatic fibrogenetic events. PMID- 16699499 TI - Complete genome sequence of the entomopathogenic and metabolically versatile soil bacterium Pseudomonas entomophila. AB - Pseudomonas entomophila is an entomopathogenic bacterium that, upon ingestion, kills Drosophila melanogaster as well as insects from different orders. The complete sequence of the 5.9-Mb genome was determined and compared to the sequenced genomes of four Pseudomonas species. P. entomophila possesses most of the catabolic genes of the closely related strain P. putida KT2440, revealing its metabolically versatile properties and its soil lifestyle. Several features that probably contribute to its entomopathogenic properties were disclosed. Unexpectedly for an animal pathogen, P. entomophila is devoid of a type III secretion system and associated toxins but rather relies on a number of potential virulence factors such as insecticidal toxins, proteases, putative hemolysins, hydrogen cyanide and novel secondary metabolites to infect and kill insects. Genome-wide random mutagenesis revealed the major role of the two-component system GacS/GacA that regulates most of the potential virulence factors identified. PMID- 16699500 TI - Short hairpin RNA-expressing bacteria elicit RNA interference in mammals. AB - RNA-interference (RNAi) is a potent mechanism, conserved from plants to humans for specific silencing of genes, which holds promise for functional genomics and gene-targeted therapies. Here we show that bacteria engineered to produce a short hairpin RNA (shRNA) targeting a mammalian gene induce trans-kingdom RNAi in vitro and in vivo. Nonpathogenic Escherichia coli were engineered to transcribe shRNAs from a plasmid containing the invasin gene Inv and the listeriolysin O gene HlyA, which encode two bacterial factors needed for successful transfer of the shRNAs into mammalian cells. Upon oral or intravenous administration, E. coli encoding shRNA against CTNNB1 (catenin beta-1) induce significant gene silencing in the intestinal epithelium and in human colon cancer xenografts in mice. These results provide an example of trans-kingdom RNAi in higher organisms and suggest the potential of bacteria-mediated RNAi for functional genomics, therapeutic target validation and development of clinically compatible RNAi-based therapies. PMID- 16699501 TI - A microengraving method for rapid selection of single cells producing antigen specific antibodies. AB - Monoclonal antibodies that recognize specific antigens of interest are used as therapeutic agents and as tools for biomedical research. Discovering a single monoclonal antibody requires retrieval of an individual hybridoma from polyclonal mixtures of cells producing antibodies with a variety of specificities. The time required to isolate hybridomas by a limiting serial-dilution, however, has restricted the diversity and breadth of available antibodies. Here we present a soft lithographic method based on intaglio printing to generate microarrays comprising the secreted products of single cells. These engraved arrays enable a rapid (<12 h) and high-throughput (>100,000 individual cells) system for identification, recovery and clonal expansion of cells producing antigen-specific antibodies. This method can be adapted, in principle, to detect any secreted product in a multiplexed manner. PMID- 16699502 TI - A global analysis of cross-talk in a mammalian cellular signalling network. AB - Cellular information processing requires the coordinated activity of a large network of intracellular signalling pathways. Cross-talk between pathways provides for complex non-linear responses to combinations of stimuli, but little is known about the density of these interactions in any specific cell. Here, we have analysed a large-scale survey of pathway interactions carried out by the Alliance for Cellular Signalling (AfCS) in RAW 264.7 macrophages. Twenty-two receptor-specific ligands were studied, both alone and in all pairwise combinations, for Ca2+ mobilization, cAMP synthesis, phosphorylation of many signalling proteins and for cytokine production. A large number of non-additive interactions are evident that are consistent with known mechanisms of cross-talk between pathways, but many novel interactions are also revealed. A global analysis of cross-talk suggests that many external stimuli converge on a relatively small number of interaction mechanisms to provide for context dependent signalling. PMID- 16699503 TI - Roles of sumoylation of a reptin chromatin-remodelling complex in cancer metastasis. AB - Defining the functional modules within transcriptional regulatory factors that govern switching between repression and activation events is a central issue in biology. Recently, we have reported the dynamic role of a beta-catenin-reptin chromatin remodelling complex in regulating a metastasis suppressor gene KAI1 (ref.1), which is capable of inhibiting the progression of tumour metastasis. Here, we identify signalling factors that confer repressive function on reptin and hence repress the expression of KAI1. Biochemical purification of a reptin containing complex has revealed the presence of specific desumoylating enzymes that reverse the sumoylation of reptin that underlies its function as a repressor. Desumoylation of reptin alters the repressive function of reptin and its association with HDAC1. Furthermore, the sumoylation status of reptin modulates the invasive activity of cancer cells with metastatic potential. These data clearly define a functional model and provide a novel link for SUMO modification in cancer metastasis. PMID- 16699504 TI - Blimp1 associates with Prmt5 and directs histone arginine methylation in mouse germ cells. AB - Blimp1, a transcriptional repressor, has a crucial role in the specification of primordial germ cells (PGCs) in mice at embryonic day 7.5 (E7.5). This SET-PR domain protein can form complexes with various chromatin modifiers in a context dependent manner. Here, we show that Blimp1 has a novel interaction with Prmt5, an arginine-specific histone methyltransferase, which mediates symmetrical dimethylation of arginine 3 on histone H2A and/or H4 tails (H2A/H4R3me2s). Prmt5 has been shown to associate with Tudor, a component of germ plasm in Drosophila melanogaster. Blimp1-Prmt5 colocalization results in high levels of H2A/H4 R3 methylation in PGCs at E8.5. However, at E11.5, Blimp1-Prmt5 translocates from the nucleus to the cytoplasm, resulting in the loss of H2A/H4 R3 methylation at the time of extensive epigenetic reprogramming of germ cells. Subsequently, Dhx38, a putative target of the Blimp1-Prmt5 complex, is upregulated. Interestingly, expression of Dhx38 is also seen in pluripotent embryonic germ cells that are derived from PGCs when Blimp1 expression is lost. Our study demonstrates that Blimp1 is involved in a novel transcriptional regulatory complex in the mouse germ-cell lineage. PMID- 16699505 TI - Opposite biases in salience-based selection for the left and right posterior parietal cortex. AB - Visual selection is determined in part by the saliency of stimuli. We assessed the brain mechanisms determining attentional responses to saliency. Repetitive transcranial magnetic stimulation (rTMS) was applied to the left and right posterior parietal cortices (PPC) immediately before participants were asked to identify a compound letter. rTMS to the right PPC disrupted the guidance of attention toward salient stimuli, whereas rTMS to the left PPC affected the ability to bias selection away from salient stimuli. We conclude that right and left PPC have opposite roles in biasing selection to and from salient stimuli in the environment. PMID- 16699506 TI - Doublecortin maintains bipolar shape and nuclear translocation during migration in the adult forebrain. AB - The ability of the mature mammalian nervous system to continually produce neuronal precursors is of considerable importance, as manipulation of this process might one day permit the replacement of cells lost as a result of injury or disease. In mammals, the anterior subventricular zone (SVZa) region is one of the primary sites of adult neurogenesis. Here we show that doublecortin (DCX), a widely used marker for newly generated neurons, when deleted in mice results in a severe morphological defect in the rostral migratory stream and delayed neuronal migration that is independent of direction or responsiveness to Slit chemorepulsion. DCX is required for nuclear translocation and maintenance of bipolar morphology during migration of these cells. Our data identifies a critical function for DCX in the movement of newly generated neurons in the adult brain. PMID- 16699507 TI - Astrocytic Ca2+ signaling evoked by sensory stimulation in vivo. AB - Although astrocytes are the most abundant cell type in the brain, evidence for their activation during physiological sensory activity is lacking. Here we show that whisker stimulation evokes increases in astrocytic cytosolic calcium (Ca(2+)) within the barrel cortex of adult mice. Increases in astrocytic Ca(2+) were a function of the frequency of stimulation, occurred within several seconds and were inhibited by metabotropic glutamate receptor antagonists. To distinguish between synaptic input and output, local synaptic activity in cortical layer 2 was silenced by iontophoresis of AMPA and NMDA receptor antagonists. The antagonists did not reduce astrocytic Ca(2+) responses despite a marked reduction in excitatory postsynaptic currents in response to whisker stimulation. These findings indicate that astrocytes respond to synaptic input, by means of spillover or ectopic release of glutamate, and that increases in astrocytic Ca(2+) occur independently of postsynaptic excitatory activity. PMID- 16699508 TI - Palmitoylation of huntingtin by HIP14 is essential for its trafficking and function. AB - Post-translational modification by the lipid palmitate is crucial for the correct targeting and function of many proteins. Here we show that huntingtin (htt) is normally palmitoylated at cysteine 214, which is essential for its trafficking and function. The palmitoylation and distribution of htt are regulated by the palmitoyl transferase huntingtin interacting protein 14 (HIP14). Expansion of the polyglutamine tract of htt, which causes Huntington disease, results in reduced interaction between mutant htt and HIP14 and consequently in a marked reduction in palmitoylation. Mutation of the palmitoylation site of htt, making it palmitoylation resistant, accelerates inclusion formation and increases neuronal toxicity. Downregulation of HIP14 in mouse neurons expressing wild-type and mutant htt increases inclusion formation, whereas overexpression of HIP14 substantially reduces inclusions. These results suggest that the expansion of the polyglutamine tract in htt results in decreased palmitoylation, which contributes to the formation of inclusion bodies and enhanced neuronal toxicity. PMID- 16699509 TI - Oncomodulin is a macrophage-derived signal for axon regeneration in retinal ganglion cells. AB - The optic nerve, like most mature CNS pathways, does not regenerate after injury. Through unknown mechanisms, however, macrophage activation in the eye stimulates retinal ganglion cells (RGCs) to regenerate long axons beyond the site of optic nerve injury. Here we identify the calcium (Ca(2+))-binding protein oncomodulin as a potent macrophage-derived growth factor for RGCs and other neurons. Oncomodulin binds to rat RGCs with high affinity in a cyclic AMP (cAMP)-dependent manner and stimulates more extensive outgrowth than other known trophic agents. Depletion of oncomodulin from macrophage-conditioned media (MCM) eliminates the axon-promoting activity of MCM. The effects of oncomodulin involve downstream signaling via Ca(2+)/calmodulin kinase and gene transcription. In vivo, oncomodulin released from microspheres promotes regeneration in the mature rat optic nerve. Oncomodulin also stimulates outgrowth from peripheral sensory neurons. Thus, oncomodulin is a new growth factor for neurons of the mature central and peripheral nervous systems. PMID- 16699510 TI - RGS4-dependent attenuation of M4 autoreceptor function in striatal cholinergic interneurons following dopamine depletion. AB - Parkinson disease is a neurodegenerative disorder whose symptoms are caused by the loss of dopaminergic neurons innervating the striatum. As striatal dopamine levels fall, striatal acetylcholine release rises, exacerbating motor symptoms. This adaptation is commonly attributed to the loss of interneuronal regulation by inhibitory D(2) dopamine receptors. Our results point to a completely different, new mechanism. After striatal dopamine depletion, D(2) dopamine receptor modulation of calcium (Ca(2+)) channels controlling vesicular acetylcholine release in interneurons was unchanged, but M(4) muscarinic autoreceptor coupling to these same channels was markedly attenuated. This adaptation was attributable to the upregulation of RGS4-an autoreceptor-associated, GTPase-accelerating protein. This specific signaling adaptation extended to a broader loss of autoreceptor control of interneuron spiking. These observations suggest that RGS4 dependent attenuation of interneuronal autoreceptor signaling is a major factor in the elevation of striatal acetylcholine release in Parkinson disease. PMID- 16699511 TI - Photocontrolled living polymerizations. AB - Living polymerizations involve the creation of polymer chains without significant irreversible chain transfer or chain termination. Such processes are widely used to access well-defined macromolecular materials with controlled architectures, such as block and star polymers. Although this concept was first realized for anionic polymerizations in the 1950s, many key recent advances have been made, most notably in the area of radical polymerization. Here, we report a living photopolymerization that involves photoexcited monomers. Exposure of metal containing ferrocenophane monomers to Pyrex-filtered light from a mercury lamp (lambda>310 nm) or to bright sunlight in the presence of an anionic initiator leads to living polymerizations, in which the conversion and molecular weight of the resulting polymer can be controlled by the irradiation time. Photoirradiation selectively weakens the iron-cyclopentadienyl bond in the monomer, allowing the use of moderately basic and highly functional-group-tolerant initiators. The polymerization proceeds through attack of the initiator and propagating anion on the iron atom of the photoexcited monomer and, remarkably, the polymerization rate decreases with increasing temperature. Block copolymer formation is possible when the light source is alternately switched on and off in between sequential addition of different monomers, providing unprecedented, photocontrolled access to new types of functional polymers. PMID- 16699512 TI - Spin gap in Tl2Ru2O7 and the possible formation of Haldane chains in three dimensional crystals. AB - Dimensionality is one of the most important parameters of physical phenomena. Only two things determine the universality class of a phase transition: the dimensionality of a given system and the symmetry of the order parameter. In most cases, the dimensionality of a substance is predetermined by its crystal structure. Examples in which the effective dimensionality is reduced are quite rare. Here we show that the three-dimensional cubic system of Tl(2)Ru(2)O(7) most probably evolves into a one-dimensional spin-one Haldane system with a spin gap below 120 K, accompanied by anomalies in the structure, resistivity and susceptibility. We argue that these anomalies are due to an orbital ordering of Ru 4d electrons, with a strong coupling among three degrees of freedom: orbital, spin and lattice. Our work provides a unique example of the spontaneous formation of Haldane system with an insight into the intriguing interplay of different degrees of freedom. PMID- 16699513 TI - The structure of the exocyst subunit Sec6p defines a conserved architecture with diverse roles. AB - The exocyst is a conserved protein complex essential for trafficking secretory vesicles to the plasma membrane. The structure of the C-terminal domain of the exocyst subunit Sec6p reveals multiple helical bundles, which are structurally and topologically similar to Exo70p and the C-terminal domains of Exo84p and Sec15, despite <10% sequence identity. The helical bundles appear to be evolutionarily related molecular scaffolds that have diverged to create functionally distinct exocyst proteins. PMID- 16699514 TI - Crystal structure of the factor XI zymogen reveals a pathway for transactivation. AB - Factor XI (FXI), a coagulation protein essential to normal hemostasis, circulates as a disulfide-linked dimer. Here we report the full-length FXI zymogen crystal structure, revealing that the protease and four apple domains assemble into a unique 'cup and saucer' architecture. The structure shows that the thrombin and platelet glycoprotein Ib binding sites are remote within the monomer but lie in close proximity across the dimer, suggesting a transactivation mechanism. PMID- 16699515 TI - Backbone and nucleobase contacts to glucosamine-6-phosphate in the glmS ribozyme. AB - The glmS ribozyme resides in the 5' untranslated region of glmS mRNA and functions as a catalytic riboswitch that regulates amino sugar metabolism in certain Gram-positive bacteria. The ribozyme catalyzes self-cleavage of the mRNA and ultimately inhibits gene expression in response to binding of glucosamine-6 phosphate (GlcN6P), the metabolic product of the GlmS protein. We have used nucleotide analog interference mapping (NAIM) and suppression (NAIS) to investigate backbone and nucleobase functional groups essential for ligand dependent ribozyme function. NAIM using GlcN6P as ligand identified requisite structural features and potential sites of ligand and/or metal ion interaction, whereas NAIS using glucosamine as ligand analog revealed those sites that orchestrate recognition of ligand phosphate. These studies demonstrate that the ligand-binding site lies in close proximity to the cleavage site in an emerging model of ribozyme structure that supports a role for ligand within the catalytic core. PMID- 16699516 TI - Dissecting Arabidopsis thaliana DICER function in small RNA processing, gene silencing and DNA methylation patterning. AB - Small RNAs have several important biological functions. MicroRNAs (miRNAs) and trans-acting small interfering RNAs (tasiRNAs) regulate mRNA stability and translation, and siRNAs cause post-transcriptional gene silencing of transposons, viruses and transgenes and are important in both the establishment and maintenance of cytosine DNA methylation. Here, we study the role of the four Arabidopsis thaliana DICER-LIKE genes (DCL1-DCL4) in these processes. Sequencing of small RNAs from a dcl2 dcl3 dcl4 triple mutant showed markedly reduced tasiRNA and siRNA production and indicated that DCL1, in addition to its role as the major enzyme for processing miRNAs, has a previously unknown role in the production of small RNAs from endogenous inverted repeats. DCL2, DCL3 and DCL4 showed functional redundancy in siRNA and tasiRNA production and in the establishment and maintenance of DNA methylation. Our studies also suggest that asymmetric DNA methylation can be maintained by pathways that do not require siRNAs. PMID- 16699517 TI - A genome-wide association study of nonsynonymous SNPs identifies a type 1 diabetes locus in the interferon-induced helicase (IFIH1) region. AB - In this study we report convincing statistical support for a sixth type 1 diabetes (T1D) locus in the innate immunity viral RNA receptor gene region IFIH1 (also known as mda-5 or Helicard) on chromosome 2q24.3. We found the association in an interim analysis of a genome-wide nonsynonymous SNP (nsSNP) scan, and we validated it in a case-control collection and replicated it in an independent family collection. In 4,253 cases, 5,842 controls and 2,134 parent-child trio genotypes, the risk ratio for the minor allele of the nsSNP rs1990760 A --> G (A946T) was 0.86 (95% confidence interval = 0.82-0.90) at P = 1.42 x 10(-10). PMID- 16699518 TI - A degradation-sensitive anionic trypsinogen (PRSS2) variant protects against chronic pancreatitis. AB - Chronic pancreatitis is a common inflammatory disease of the pancreas. Mutations in the genes encoding cationic trypsinogen (PRSS1) and the pancreatic secretory trypsin inhibitor (SPINK1) are associated with chronic pancreatitis. Because increased proteolytic activity owing to mutated PRSS1 enhances the risk for chronic pancreatitis, mutations in the gene encoding anionic trypsinogen (PRSS2) may also predispose to disease. Here we analyzed PRSS2 in individuals with chronic pancreatitis and controls and found, to our surprise, that a variant of codon 191 (G191R) is overrepresented in control subjects: G191R was present in 220/6,459 (3.4%) controls but in only 32/2,466 (1.3%) affected individuals (odds ratio 0.37; P = 1.1 x 10(-8)). Upon activation by enterokinase or trypsin, purified recombinant G191R protein showed a complete loss of trypsin activity owing to the introduction of a new tryptic cleavage site that renders the enzyme hypersensitive to autocatalytic proteolysis. In conclusion, the G191R variant of PRSS2 mitigates intrapancreatic trypsin activity and thereby protects against chronic pancreatitis. PMID- 16699519 TI - Stochastic kinetics of intracellular huntingtin aggregate formation. AB - Neurodegeneration in Huntington disease is described by neuronal loss in which the probability of cell death remains constant with time. However, the quantitative connection between the kinetics of cell death and the molecular mechanism initiating neurodegeneration remains unclear. One hypothesis is that nucleation of protein aggregates containing exon I fragments of the mutant huntingtin protein (mhttex1), which contains an expanded polyglutamine region in patients with the disease, is the explanation for the infrequent but steady occurrence of neuronal death, resulting in adult onset of the disease. Recent in vitro evidence suggests that sufficiently long polyglutamine peptides undergo a unimolecular conformational change to form a nucleus that seeds aggregation. Here we use this nucleation mechanism as the basis to derive a stochastic mathematical model describing the probability of aggregate formation in cells as a function of time and mhttex1 protein concentration, and validate the model experimentally. These findings suggest that therapeutic strategies for Huntington disease predicated on reducing the rate of mhttex1 aggregation need only make modest reductions in huntingtin expression level to substantially increase the delay time until aggregate formation. PMID- 16699520 TI - The nongenotoxic carcinogens naphthalene and para-dichlorobenzene suppress apoptosis in Caenorhabditis elegans. AB - Naphthalene (1) and para-dichlorobenzene (PDCB, 2), which are widely used as moth repellents and air fresheners, cause cancer in rodents and are potential human carcinogens. However, their mechanisms of action remain unclear. Here we describe a novel method for delivering and screening hydrophobic chemicals in C. elegans and apply this technique to investigate the ways in which naphthalene and PDCB may promote tumorigenesis in mammals. We show that naphthalene and PDCB inhibit apoptosis in C. elegans, a result that suggests a cellular mechanism by which these chemicals may promote the survival and proliferation of latent tumor cells. In addition, we find that a naphthalene metabolite directly inactivates caspases by oxidizing the active site cysteine residue; this suggests a molecular mechanism by which these chemicals suppress apoptosis. Naphthalene and PDCB are the first small-molecule apoptosis inhibitors identified in C. elegans. The power of C. elegans molecular genetics, in combination with the possibility of carrying out large-scale chemical screens in this organism, makes C. elegans an attractive and economic animal model for both toxicological studies and drug screens. PMID- 16699521 TI - Isolation of a novel acidiphilic methanogen from an acidic peat bog. AB - Acidic peatlands are among the largest natural sources of atmospheric methane and harbour a large diversity of methanogenic Archaea. Despite the ubiquity of methanogens in these peatlands, indigenous methanogens capable of growth at acidic pH values have resisted culture and isolation; these recalcitrant methanogens include members of an uncultured family-level clade in the Methanomicrobiales prevalent in many acidic peat bogs in the Northern Hemisphere. However, we recently succeeded in obtaining a mixed enrichment culture of a member of this clade. Here we describe its isolation and initial characterization. We demonstrate that the optimum pH for methanogenesis by this organism is lower than that of any previously described methanogen. PMID- 16699522 TI - Single-cell proteomic analysis of S. cerevisiae reveals the architecture of biological noise. AB - A major goal of biology is to provide a quantitative description of cellular behaviour. This task, however, has been hampered by the difficulty in measuring protein abundances and their variation. Here we present a strategy that pairs high-throughput flow cytometry and a library of GFP-tagged yeast strains to monitor rapidly and precisely protein levels at single-cell resolution. Bulk protein abundance measurements of >2,500 proteins in rich and minimal media provide a detailed view of the cellular response to these conditions, and capture many changes not observed by DNA microarray analyses. Our single-cell data argue that noise in protein expression is dominated by the stochastic production/destruction of messenger RNAs. Beyond this global trend, there are dramatic protein-specific differences in noise that are strongly correlated with a protein's mode of transcription and its function. For example, proteins that respond to environmental changes are noisy whereas those involved in protein synthesis are quiet. Thus, these studies reveal a remarkable structure to biological noise and suggest that protein noise levels have been selected to reflect the costs and potential benefits of this variation. PMID- 16699523 TI - Live imaging of yeast Golgi cisternal maturation. AB - There is a debate over how protein trafficking is performed through the Golgi apparatus. In the secretory pathway, secretory proteins that are synthesized in the endoplasmic reticulum enter the early compartment of the Golgi apparatus called cis cisternae, undergo various modifications and processing, and then leave for the plasma membrane from the late (trans) cisternae. The cargo proteins must traverse the Golgi apparatus in the cis-to-trans direction. Two typical models propose either vesicular transport or cisternal progression and maturation for this process. The vesicular transport model predicts that Golgi cisternae are distinct stable compartments connected by vesicular traffic, whereas the cisternal maturation model predicts that cisternae are transient structures that form de novo, mature from cis to trans, and then dissipate. Technical progress in live-cell imaging has long been awaited to address this problem. Here we show, by the use of high-speed three-dimensional confocal microscopy, that yeast Golgi cisternae do change the distribution of resident membrane proteins from the cis nature to the trans over time, as proposed by the maturation model, in a very dynamic way. PMID- 16699524 TI - Golgi maturation visualized in living yeast. AB - The Golgi apparatus is composed of biochemically distinct early (cis, medial) and late (trans, TGN) cisternae. There is debate about the nature of these cisternae. The stable compartments model predicts that each cisterna is a long-lived structure that retains a characteristic set of Golgi-resident proteins. In this view, secretory cargo proteins are transported by vesicles from one cisterna to the next. The cisternal maturation model predicts that each cisterna is a transient structure that matures from early to late by acquiring and then losing specific Golgi-resident proteins. In this view, secretory cargo proteins traverse the Golgi by remaining within the maturing cisternae. Various observations have been interpreted as supporting one or the other mechanism. Here we provide a direct test of the two models using three-dimensional time-lapse fluorescence microscopy of the yeast Saccharomyces cerevisiae. This approach reveals that individual cisternae mature, and do so at a consistent rate. In parallel, we used pulse-chase analysis to measure the transport of two secretory cargo proteins. The rate of cisternal maturation matches the rate of protein transport through the secretory pathway, suggesting that cisternal maturation can account for the kinetics of secretory traffic. PMID- 16699525 TI - Negative regulation of interferon-regulatory factor 3-dependent innate antiviral response by the prolyl isomerase Pin1. AB - Recognition of double-stranded RNA activates interferon-regulatory factor 3 (IRF3)-dependent expression of antiviral factors. Although the molecular mechanisms underlying the activation of IRF3 have been studied, the mechanisms by which IRF3 activity is reduced have not. Here we report that activation of IRF3 is negatively regulated by the peptidyl-prolyl isomerase Pin1. After stimulation by double-stranded RNA, induced phosphorylation of the Ser339-Pro340 motif of IRF3 led to its interaction with Pin1 and finally polyubiquitination and then proteasome-dependent degradation of IRF3. Suppression of Pin1 by RNA interference or genetic deletion resulted in enhanced IRF-3-dependent production of interferon beta, with consequent reduction of virus replication. These results elucidate a previously unknown mechanism for controlling innate antiviral responses by negatively regulating IRF3 activity via Pin1. PMID- 16699526 TI - Regulation of T lymphopoiesis by Notch1 and Lunatic fringe-mediated competition for intrathymic niches. AB - Notch1 activation regulates T lineage commitment and early T cell development. Fringe glycosyltransferases alter the sensitivity of Notch receptors to Delta like versus Jagged Notch ligands, but their functions in T lymphopoiesis have not been defined. Here we show that developmental stage-specific expression of the glycosyltransferase lunatic fringe (Lfng) is required for coordination of the access of T cell progenitors to intrathymic niches that support Notch1-dependent phases of T cell development. Lfng-null progenitors generated few thymocytes in competitive assays, whereas Lfng overexpression converted thymocytes into 'supercompetitors' with enhanced binding of Delta-like ligands and blocked T lymphopoiesis from normal progenitors. We suggest that the ability of Lfng and Notch1 to control progenitor competition for limiting cortical niches is an important mechanism for the homeostatic regulation of thymus size. PMID- 16699527 TI - Prolonged responses after autologous stem cell transplantation in African American patients with multiple myeloma. AB - Multiple myeloma (MM) has a double incidence in African-American (AA) than in non AA patients and previous studies have shown a higher mortality in the former patient population. Here, we retrospectively analyzed the results of autologous stem cell transplantation (ASCT) in 38 AA and 32 non-AA consecutive patients. The two groups were comparable at diagnosis for age, stage of the disease, cytogenetic abnormalities, beta(2) microglobulin and albumin blood levels, and plasma cell marrow infiltration. The rates of complete and partial response observed in AA and non-AA patients after induction chemotherapy (9 and 42 vs 13 and 33%) and at 2 months (31 and 25 vs 30 and 20%) following ASCT were similar. At 6 months after ASCT, a greater relapse rate was observed in non-AA patients (P=0.009). At a median follow-up of 26 months, AA patients had a greater event free survival (P=0.02) than non-AA patients, whereas overall survival was comparable in the two groups. The initial finding that AA patients with MM, compared to non-AA patients, had more prolonged responses and comparable survival after ASCT suggests that intensified chemotherapy is equally effective in patients of various ethnicities. PMID- 16699528 TI - The modification of high-dose therapy shortens the duration of neutropaenia by delay of leucocyte nadir. AB - Infections during neutropaenia contribute still significantly to mortality and morbidity after high-dose therapy and autologous stem cell transplantation. Further acceleration of haemopoietic recovery seems impossible for biological reasons. Another approach to shorten neutropaenia could be to remove drugs from high-dose therapy protocols with strong contribution to immunosuppression and neutropaenia and unproven antineoplastic activity. In this retrospective matched pair analysis, conventional busulphan/cyclophosphamide (Bu/Cy) high-dose therapy was compared to single-agent busulphan conditioning before autologous stem cell transplantation. This modification led to a significant shorter neutropaenic interval by protraction of cell decrease and to a significant mitigation of neutropaenia. After single-agent busulphan conditioning, leucocytes dropped below 1/nl at median 1.5 days later when compared to the patients from the busulphanBu/Cy control group (P=0.001). In a significant percentage of patients (n=6, 60%) leucocytes did not fall below 0.5 cells/nl at any time. In contrast, all patients from the Bu/Cy control group experienced deep neutropaenia (P=0.004). Thrombocytopaenia and requirement for transfusions of platelets or red cells were not influenced. Antineoplastic activity seemed to be preserved as determined by survival analysis. In conclusion, modification of high-dose regimen with the intention to shorten neutropaenia with preserved antitumour activity could be an approach to reduce infection-related morbidity and mortality and to consider economic necessities. PMID- 16699529 TI - Description of an adenovirus A31 outbreak in a paediatric haematology unit. AB - Adenovirus infections result in significant morbidity and mortality in allogeneic haematopoietic stem cell transplanted (hSCT) children. Adenovirus from species C and B account for more than 90% of adenoviruses recovered after hSCT. However, infections due to adenovirus A31 have been increasingly reported in recent years. Between April 2002 and April 2005, blood samples obtained every 2 weeks from 58 hSCT children were screened for adenovirus species A to C by quantitative real time PCR. Phylogenetic analysis was realized after amplification and sequencing of the entire hexon gene. Fifteen cases of adenovirus infection with viraemia were recovered during this 3 years period. During spring/summer 2003, seven cases occurred and were due to an adenovirus species A. Phylogenetic analysis of the seven strains showed that they belonged to the A31 genotype and shared 100% homology. Clinical features of the seven HSCT children with A31 adenovirus viraemia are described. We describe here an epidemic spread of adenovirus genotype A31 in a paediatric haematology unit. Timing, location and hexon gene genotyping results highly suggested a nosocomial origin to this epidemic. The burden of adenovirus A31 infection needs to be further assessed in this context. PMID- 16699530 TI - Rapid helper T-cell recovery above 200 x 10 6/l at 3 months correlates to successful transplant outcomes after allogeneic stem cell transplantation. AB - The current study evaluates the role of quantitative measurement of peripheral lymphocyte subsets, especially CD4+ helper T-cell recovery, in predicting transplant outcomes including overall survival (OS) and non-relapse mortality (NRM) after allogeneic stem cell transplantation. A total of 69 allogeneic recipients were included with following diagnoses: acute myeloid leukemia 42, acute lymphoblastic leukemia 5, chronic myeloid leukemia 15, non-Hodgkin's lymphoma 5 and high-risk myelodysplastic syndrome 2. The peripheral lymphocyte subset counts (CD3+ T cells, CD3+4+ helper T cells, CD3+8+ cytotoxic T cells, CD19+ B cells, and CD56+ natural killer cells) were measured at 3, 6 and 12 months. The CD4+ helper T-cell reconstitution at 3 months was strongly correlated with OS (P<0.0001), NRM (P=0.0007), and opportunistic infections (P=0.0108) at the cutoff value of 200 x 10(6)/l CD4(+) helper T cells. Rapid CD4+ helper T-cell recovery was also associated with a higher CD4+ helper T-cell transplant dose (P=0.006) and donor type (P<0.001). An early CD4+ helper T-cell recovery at 3 months correlated with a subsequent faster helper T-cell recovery until 12 months, yet not with B-cell recovery. In a multivariate analysis, rapid recovery of CD4+ helper T cells at 3 months was a favorable prognostic factor together with higher CD34+ cell transplant dose in terms of OS (P=0.001) and NRM (P=0.005). PMID- 16699531 TI - Combination antithymocyte globulin and soluble TNFalpha inhibitor (etanercept) +/ mycophenolate mofetil for treatment of steroid refractory acute graft-versus host disease. AB - Antitumor necrosis factor-alpha antibodies are increasingly being used for the treatment of steroid-refractory acute graft-versus-host disease (GVHD) complicating allogeneic stem cell transplantation. We retrospectively reviewed the outcomes of 16 patients with refractory acute predominantly visceral GVHD treated with combination antithymocyte globulin (ATG), tacrolimus and etanercept +/- mycophenolate mofetil (MMF) at our institution. Overall response rate (CR+PR) was 81%, with median survival post commencing salvage immunosuppression 224 days (range 20-1216 days). In total, eight patients (50%) died, including from progressive GVHD in two cases (13%), infection in five (31%) and relapse of underlying malignancy in one (6%). In comparison to our previous experience of ATG+tacrolimus as treatment for refractory visceral GVHD, both response rate and overall survival were improved with addition of etanercept, with no apparent increase in infectious complications. As such, use of etanercept in combination with ATG +/- MMF for treatment of steroid refractory acute GVHD appears to be associated with high response rates, significant survival and no unexpected toxicity. Further study of this immunosuppression combination in a larger cohort of patients in this setting is indicated. PMID- 16699532 TI - Response to high-dose dexamethasone for acquired pure red cell aplasia following ABO-mismatched allogeneic stem cell transplantation. PMID- 16699533 TI - Autologous stem cell transplantation in patients with chronic lymphocytic leukaemia: the Finnish experience. AB - Although autologous stem cell transplantation (ASCT) has gained some popularity as a treatment option in patients with chronic lymphocytic leukaemia (CLL), limited multicentre data are available on the feasibility and efficacy of this approach. Between January 1995 and June 2005, 72 patients with CLL received ASCT in five Finnish centres. There were 45 men and 27 women with a median age of 57 years (38-69). The median time from diagnosis to ASCT was 32 months (6-181) and the median number of prior regimens 1 (1-4). All patients received blood stem cell grafts and CD34+ selection had been performed in 44 patients (61%). The most common high-dose regimen was a total body irradiation plus cyclophosphamide (38 patients, 53%). No early treatment-related deaths were observed. With a median follow-up of 28 months from ASCT, a relapse or progression has been observed in 27 patients (37%). The projected progression-free survival is 48 months (confidence interval (CI) 30-66). The projected median overall survival is 95 months (CI 74-101) from ASCT and is not influenced by graft selection or conditioning regimen used. Autologous stem cell transplantation is a feasible treatment option for CLL. Randomized trials against alternative treatments are needed to assess the impact of ASCT on the clinical course of CLL. PMID- 16699534 TI - Endocrine dysfunction and parameters of the metabolic syndrome after bone marrow transplantation during childhood and adolescence. AB - Endocrine dysfunction and parameters of metabolic syndrome were assessed in 91 patients aged 4.3-32.5 years who underwent allogeneic or autologous BMT in childhood. Final short stature, found in five of the 35 patients who attained final height, was associated with the underlying disease (specifically, Fanconi anemia) (P=0.0013), previous cranial irradiation (P=0.0007), type of conditioning irradiation (P<0.05) and allogeneic BMT (P=0.05). Growth hormone deficiency (n=10) was associated with previous cranial irradiation (P<0.005) and conditioning total body irradiation (P<0.001). Twelve patients had primary hypothyroidism, one had hyperthyroidism and one papillary thyroid carcinoma. Hypothyroidism was associated with neck/mediastinal (P<0.005) and conditioning irradiation (P<0.05). Primary gonadal failure was found in 24 of the mature patients (62.5% females). Hypogonadism was associated with the underlying disease (especially hematological malignancies) (P<0.05), pretransplant treatment (P<0.05), irradiation conditioning (P<0.001), older age (P<0.005) and advanced pubertal stage at BMT (P<0.05). Obesity (body mass index >2 s.d.) was found in 4.4% and type II diabetes and impaired glucose tolerance in 3.3% each. Dyslipidemia was found in 27.9% of the 43 patients tested. These findings emphasize the need for long-term follow-up of endocrine and metabolic parameters in young patients after BMT in order to offer proper treatment and improve quality of life. PMID- 16699536 TI - Double-stranded DNA breaks and gene functions in recombination and meiosis. AB - Meiotic prophase I is a long and complex phase. Homologous recombination is an important process that occurs between homologous chromosomes during meiotic prophase I. Formation of chiasmata, which hold homologous chromosomes together until the metaphase I to anaphase I transition, is critical for proper chromosome segregation. Recent studies have suggested that the SPO11 proteins have conserved functions in a number of organisms in generating sites of double-stranded DNA breaks (DSBs) that are thought to be the starting points of homologous recombination. Processing of these sites of DSBs requires the function of RecA homologs, such as RAD51, DMC1, and others, as suggested by mutant studies; thus the failure to repair these meiotic DSBs results in abnormal chromosomal alternations, leading to disrupted meiosis. Recent discoveries on the functions of these RecA homologs have improved the understanding of the mechanisms underlying meiotic homologous recombination. PMID- 16699537 TI - Ubiquitination-mediated protein degradation and modification: an emerging theme in plant-microbe interactions. AB - Post-translational modification is central to protein stability and to the modulation of protein activity. Various types of protein modification, such as phosphorylation, methylation, acetylation, myristoylation, glycosylation, and ubiquitination, have been reported. Among them, ubiquitination distinguishes itself from others in that most of the ubiquitinated proteins are targeted to the 26S proteasome for degradation. The ubiquitin/26S proteasome system constitutes the major protein degradation pathway in the cell. In recent years, the importance of the ubiquitination machinery in the control of numerous eukaryotic cellular functions has been increasingly appreciated. Increasing number of E3 ubiquitin ligases and their substrates, including a variety of essential cellular regulators have been identified. Studies in the past several years have revealed that the ubiquitination system is important for a broad range of plant developmental processes and responses to abiotic and biotic stresses. This review discusses recent advances in the functional analysis of ubiquitination-associated proteins from plants and pathogens that play important roles in plant-microbe interactions. PMID- 16699538 TI - The brassinosteroid signal transduction pathway. AB - Steroids function as signaling molecules in both animals and plants. While animal steroid hormones are perceived by nuclear receptor family of transcription factors, brassinosteroids (BR) in plants are perceived by a cell surface receptor kinase, BRI1. Recent studies have demonstrated that BR binding to the extracellular domain of BRI1 induces kinase activation and dimerization with another receptor kinase, BAK1. Activated BRI1 or BAK1 then regulate, possibly indirectly, the activities of BIN2 kinase and/or BSU1 phosphatase, which directly regulate the phosphorylation status and nuclear accumulation of two homologous transcription factors, BZR1 and BES1. BZR1 and BES1 directly bind to promoters of BR responsive genes to regulate their expression. The BR signaling pathway has become a paradigm for both receptor kinase signaling in plants and steroid signaling by cell surface receptors in general. PMID- 16699539 TI - A RHOse by any other name: a comparative analysis of animal and plant Rho GTPases. AB - Rho GTPases are molecular switches that act as key regulators of a many cellular processes, including cell movement, morphogenesis, host defense, cell division and gene expression. Rho GTPases are found in all eukaryotic kingdoms. Plants lack clear homologs to conventional Rho GTPases found in yeast and animals; instead, they have over time developed a unique subfamily, ROPs, also known as RAC. The origin of ROP-like proteins appears to precede the appearance of land plants. This review aims to discuss the evolution of ROP/RAC and to compare plant ROP and animal Rho GTPases, focusing on similarities and differences in regulation of the GTPases and their downstream effectors. PMID- 16699540 TI - BUD2, encoding an S-adenosylmethionine decarboxylase, is required for Arabidopsis growth and development. AB - Polyamines are implicated in regulating various developmental processes in plants, but their exact roles and how they govern these processes still remain elusive. We report here an Arabidopsis bushy and dwarf mutant, bud2, which results from the complete deletion of one member of the small gene family that encodes S-adenosylmethionine decarboxylases (SAMDCs) necessary for the formation of the indispensable intermediate in the polyamine biosynthetic pathway. The bud2 plant has enlarged vascular systems in inflorescences, roots, and petioles, and an altered homeostasis of polyamines. The double mutant of bud2 and samdc1, a knockdown mutant of another SAMDC member, is embryo lethal, demonstrating that SAMDCs are essential for plant embryogenesis. Our results suggest that polyamines are required for the normal growth and development of higher plants. PMID- 16699541 TI - Plant fertility defects induced by the enhanced expression of microRNA167. AB - The plant hormone auxin plays a critical role in regulating plant growth and development. Recent advances have been made in the understanding of auxin response pathways, primarily by the characterization of auxin response mutants in Arabidopsis. In addition, microRNAs (miRNAs) have been shown to be critical regulators of genes important for normal plant development and physiology. However, little is known about possible interactions between miRNAs and hormonal signaling during normal development. Here we show that an Arabidopsis microRNA, miR167, which has a complementary sequence to a portion of the AUXIN RESPONSE FACTOR6 (ARF6) and ARF8 mRNAs, can cause transcript degradation for ARF8, but not for ARF6. We report phenotypic characterizations of 35S::MIR167b transgenic lines, and show that severe 35S::MIR167b transgenic lines had phenotypes similar to those of an arf6 arf8 double mutant. The transgenic phenotypes suggest that miR167 may repress ARF6 at the level of translation. We demonstrate that the transgenic plants are defective in all four whorls of floral organs. In the transgenic flowers, filaments were abnormally short, anthers could not properly release pollen, and pollen grains did not germinate. Our results provide an important link between the miRNA-mediated regulatory pathway of gene expression and the auxin signaling network promoting plant reproductive development. PMID- 16699542 TI - MORN motifs in plant PIPKs are involved in the regulation of subcellular localization and phospholipid binding. AB - Multiple repeats of membrane occupation and recognition nexus (MORN) motifs were detected in plant phosphatidylinositl monophosphate kinase (PIPK), a key enzyme in PI-signaling pathway. Structural analysis indicates that all the MORN motifs (with varied numbers at ranges of 7-9), which shared high homologies to those of animal ones, were located at N-terminus and sequentially arranged, except those of OsPIPK1 and AtPIPK7, in which the last MORN motif was separated others by an approximately 100 amino-acid "island" region, revealing the presence of two kinds of MORN arrangements in plant PIPKs. Through employing a yeast-based SMET (sequence of membrane-targeting) system, the MORN motifs were shown being able to target the fusion proteins to cell plasma membrane, which were further confirmed by expression of fused MORN-GFP proteins. Further detailed analysis via deletion studies indicated the MORN motifs in OsPIPK1, together with the 104 amino-acid "island" region are involved in the regulation of differential subcellular localization, i.e. plasma membrane or nucleus, of the fused proteins. Fat Western blot analysis of the recombinant MORN polypeptide, expressed in Escherichia coli, showed that MORN motifs could strongly bind to PA and relatively slightly to PI4P and PI(4,5)P2. These results provide informative hints on mechanisms of subcellular localization, as well as regulation of substrate binding, of plant PIPKs. PMID- 16699544 TI - Proteomic identification of potential target proteins regulated by an ASK1 mediated proteolysis pathway. AB - The ASK1 (ARABIDOPSIS SKP1-LIKE) protein is a critical component of the SCF (Skp1 Cullin-F box protein) ubiquitin ligase complexes that recruit target proteins for degradation by the 26S proteosome. To investigate proteins that are affected by the ASK1-mediated proteolysis pathway in Arabidopsis flowers, we compared the proteomes of the Arabidopsis wild type and ask1 mutant flower buds using two dimensional electrophoresis (2-DE). Ten protein spots with higher or lower abundance in the ask1 mutant flowers compared to wild type flowers were excised and subjected to further mass spectrometry (MS) analysis. The results showed that they were proteins involved in photomorphogenesis, circadian oscillation, post translation process, stress-responses and cell expansion or elongation, suggesting that those processes were affected in the ask1 mutant. The transcript levels of these genes were also compared based on the Affymetrix gene chip microarray data. No significant difference was observed for most of the genes, suggesting that the proteins with elevated levels of accumulation in the ask1 mutant could be candidate targets regulated by an ASK1-mediated proteolysis pathway. These results help to elucidate the pleiotropic functions of ASK1 in Arabidopsis developmental processes and also demonstrate the importance and necessity of studying protein levels with respect to gene functions. PMID- 16699543 TI - Arabidopsis thaliana histone deacetylase 1 (AtHD1) is localized in euchromatic regions and demonstrates histone deacetylase activity in vitro. AB - Arabidopsis thaliana histone deacetylase 1 (AtHD1 or AtHDA19), a homolog of yeast RPD3, is a global regulator of many physiological and developmental processes in plants. In spite of the genetic evidence for a role of AtHD1 in plant gene regulation and development, the biochemical and cellular properties of AtHD1 are poorly understood. Here we report cellular localization patterns of AtHD1 in vivo and histone deacetylase activity in vitro. The transient and stable expression of a green fluorescent protein (GFP)-tagged AtHD1 in onion cells and in roots, seeds and leaves of the transgenic Arabidopsis, respectively, revealed that AtHD1 is localized in the nucleus presumably in the euchromatic regions and excluded from the nucleolus. The localization patterns of AtHD1 are different from those of AtHD2 and AtHDA6 that are involved in nucleolus formation and silencing of transgenes and repeated DNA elements, respectively. In addition, a histone deacetylase activity assay showed that the recombinant AtHD1 produced in bacteria demonstrated a specific histone deacetylase activity in vitro. The data suggest that AtHD1 is a nuclear protein and possesses histone deacetylase activities responsible for global transcriptional regulation important to plant growth and development. PMID- 16699545 TI - Wrinkled petals and stamens 1, is required for the morphogenesis of petals and stamens in Lotus japonicus. AB - Although much progress has been made in understanding how floral organ identity is determined during the floral development, less is known about how floral organ is elaborated in the late floral developmental stages. Here we describe a novel floral mutant, wrinkled petals and stamens1 (wps1), which shows defects in the development of petals and stamens. Genetic analysis indicates that wps1 mutant is corresponding to a single recessive locus at the long arm of chromosome 3. The early development of floral organs in wps1 mutant is similar to that in wild type, and the malfunction of the mutant commences in late developmental stages, displaying a defect on the appearance of petals and stamens. In the mature flower, petals and stamen filaments in the mutant are wrinkled or folded, and the cellular morphology under L1 layer of petals and stamen filaments is abnormal. It is found that the expression patterns of floral organ identity genes are not affected in wps1 mutants compared with that of wild type, consistent with the unaltered development of all floral organs. Furthermore, the identities of epidermal cells in different type of petals are maintained. The histological analysis shows that in wps1 flowers all petals are irregularly folded, and there are knotted structures in the petals, while the shape and arrangement of inner cells are malformed and unorganized. Based on these results, we propose that Wps1 acts downstream to the class B floral organ identity genes, and functions to modulate the cellular differentiation during the late flower developmental stages. PMID- 16699546 TI - Duplication and expression analysis of multicopy miRNA gene family members in Arabidopsis and rice. AB - To understand the expansion of multicopy microRNA (miRNA) families in plants, we localized the reported miRNA genes from Arabidopsis and rice to their chromosomes, respectively, and observed that 37% of 117 miRNA genes from Arabidopsis and 35% of 173 miRNA genes from rice were segmental duplications in the genome. In order to characterize whether the expression diversification has occurred among plant multicopy miRNA family members, we designed PCR primers targeting 48 predicted miRNA precursors from 10 families in Arabidopsis and rice. Results from RT-PCR data suggest that the transcribed precursors of members within the same miRNA family were present at different expression levels. In addition, although miR160 and miR162 sequences were conserved in Arabidopsis and rice, we found that the expression patterns of these genes differed between the two species. These data suggested that expression diversification has occurred in multicopy miRNA families, increasing our understanding of the expression regulation of miRNAs in plants. PMID- 16699547 TI - Development of rabbit monoclonal and polyclonal antibodies for detection of site specific histone modifications and their application in analyzing overall modification levels. AB - In addition to DNA sequence information, site-specific histone modifications are another important determinant of gene expression in a eukaryotic organism. We selected four modification sites in common histones that are known to significantly impact chromatin function and generated monoclonal or polyclonal antibodies that recognize each of those site-specific modifications. We used these antibodies to demonstrate that the site-specific histone modification levels remain relatively constant in different organs of the same organism. We also compared the levels of selected histone modifications among several representative organisms and found that site-specific modifications are highly variable among different organisms, providing new insight into the evolutionary divergence of specific histone modifications. PMID- 16699548 TI - Characterization of chromosome-specific S-SAP markers and their use in studying genetic diversity in Aegilops species. AB - The short interspersed nuclear element (SINE), Au, was used to develop sequence specific amplified polymorphism (S-SAP) markers for U- and M-genome chromosomes. The markers were localized using Triticum aestivum (wheat)-- Aegilops geniculata and wheat-- Aegilops biuncialis disomic chromosome addition lines. Thirty-seven markers distributed over 6 U and 6 M chromosomes were produced. A genetic diversity study carried out on 37 accessions from Ae. biuncialis, Ae. comosa, Ae. geniculata, and Ae. umbellulata suggested that Ae. biuncialis have arisen from its diploid ancestors more recently than Ae. geniculata. Several earlier studies indicated that the M genomes in polyploid Aegilops species had accumulated substantial rearrangements, whereas the U genomes remained essentially unmodified. However, this cannot be attributed to the preferential insertion of retroelements into the M genome chromosomes. Fourteen markers from a total of 8 chromosomes were sequenced; 3 markers were similar to known plant genes, 1 was derived from a long terminal repeat (LTR) retrotransposon, and 10 markers did not match to any known DNA sequences, suggesting that they were located in the highly variable intergenic regions. PMID- 16699549 TI - Quantification of genetic relationships among A genomes of wheats. AB - The genetic relationships of A genomes of Triticum urartu (Au) and Triticum monococcum (Am) in polyploid wheats are explored and quantified by AFLP fingerprinting. Forty-one accessions of A-genome diploid wheats, 3 of AG-genome wheats, 19 of AB-genome wheats, 15 of ABD-genome wheats, and 1 of the D-genome donor Ae. tauschii have been analysed. Based on 7 AFLP primer combinations, 423 bands were identified as potentially A genome specific. The bands were reduced to 239 by eliminating those present in autoradiograms of Ae. tauschii, bands interpreted as common to all wheat genomes. Neighbour-joining analysis separates T. urartu from T. monococcum. Triticum urartu has the closest relationship to polyploid wheats. Triticum turgidum subsp. dicoccum and T. turgidum subsp. durum lines are included in tightly linked clusters. The hexaploid spelts occupy positions in the phylogenetic tree intermediate between bread wheats and T. turgidum. The AG-genome accessions cluster in a position quite distant from both diploid and other polyploid wheats. The estimates of similarity between A genomes of diploid and polyploid wheats indicate that, compared with Am, Au has around 20% higher similarity to the genomes of polyploid wheats. Triticum timo pheevii AG genome is molecularly equidistant from those of Au and Am wheats. PMID- 16699550 TI - Analysis of ESTs from multiple Gossypium hirsutum tissues and identification of SSRs. AB - In an effort to expand the Gossypium hirsutum L. (cotton) expressed sequence tag (EST) database, ESTs representing a variety of tissues and treatments were sequenced. Assembly of these sequences with ESTs already in the EST database (dbEST, GenBank) identified 9675 cotton sequences not present in GenBank. Statistical analysis of a subset of these ESTs identified genes likely differentially expressed in stems, cotyledons, and drought-stressed tissues. Annotation of the differentially expressed cDNAs tentatively identified genes involved in lignin metabolism, starch biosynthesis and stress response, consistent with pathways likely to be active in the tissues under investigation. Simple sequence repeats (SSRs) were identified among these ESTs, and an inexpensive method was developed to screen genomic DNA for the presence of these SSRs. At least 69 SSRs potentially useful in mapping were identified. Selected amplified SSRs were isolated and sequenced. The sequences corresponded to the EST containing the SSRs, confirming that these SSRs will potentially map the gene represented by the EST. The ESTs containing SSRs were annotated to help identify the genes that may be mapped using these markers. PMID- 16699551 TI - A genetic linkage map of Silene vulgaris based on AFLP markers. AB - A genetic linkage map of an intraspecific cross between 2 Silene vulgaris s.l. ecotypes is presented. Three-hundred AFLP markers from 2 different restriction enzyme combinations were used to genotype an F2 mapping population. Maternal and paternal pure-coupling phase maps with 114 and 186 markers on 12 and 13 linkage groups, respectively, were constructed. Total map length of the paternal and maternal maps are 547 and 446 Kosambi cM, respectively. Nearly half of the markers (49%) exhibited significant transmission ratio distortion. Genome coverage and potential causes of the observed segregation ratio distortions are discussed. The maps represent a first step towards the identification of quantitative trait loci associated with habitat adaptation in the non-model species Silene vulgaris. PMID- 16699552 TI - Genetic diversity in colonial bentgrass (Agrostis capillaris L.) revealed by EcoRI-MseI and PstI-MseI AFLP markers. AB - Colonial bentgrass (Agrostis capillaris L.) is a potential source for genetic improvement of resistance to environmental stress and disease for other bentgrass species (Agrostis spp.). To conserve and study the existing genetic resources of colonial bentgrass for use in breeding, genetic diversity was investigated using amplified fragment length polymorphism (AFLP) markers. Included in this study were 22 accessions from US Department of Agriculture germplasm collected from 11 countries, in conjunction with 14 accessions from northern Spain and 3 commercial cultivars. Ten EcoRI-MseI and 6 PstI-MseI AFLP primer combinations produced 181 and 128 informative polymorphic bands, respectively. Cluster analysis of genetic similarity estimates revealed a high level of diversity in colonial bentgrass species with averages of 0.51 (EcoRI-MseI) and 0.63 (PstI-MseI). Greater genetic diversity was detected by the EcoRI-MseI AFLP primer combinations. A low but significant positive correlation (r = 0.44, p = 0.0099) between the 2 Jaccard similarity matrices was obtained by the Mantel test. Commercial cultivars of bentgrass showed a narrow genetic background. The assessment of genetic diversity among colonial bentgrass accessions suggested the potential value of the colonial bentgrass germplasm in turfgrass cultivar improvement. PMID- 16699553 TI - Chromosome structural changes in diploid and tetraploid A genomes of Gossypium. AB - The genus Gossypium, which comprises a divergent group of diploid species and several recently formed allotetraploids, offers an excellent opportunity to study polyploid genome evolution. In this study, chromosome structural variation among the A, At, and D genomes of Gossypium was evaluated by comparative genetic linkage mapping. We constructed a fully resolved RFLP linkage map for the diploid A genome consisting of 275 loci using an F2 interspecific Gossypium arboreum x Gossypium herbaceum family. The 13 chromosomes of the A genome are represented by 12 large linkage groups in our map, reflecting an expected interchromosomal translocation between G. arboreum and G. herbaceum. The A-genome chromosomes are largely collinear with the D genomes, save for a few small inversions. Although the 2 diploid mapping parents represent the closest living relatives of the allotetraploid At-genome progenitor, 2 translocations and 7 inversions were observed between the A and At genomes. The recombination rates are similar between the 2 diploid genomes; however, the At genome shows a 93% increase in recombination relative to its diploid progenitors. Elevated recombination in the Dt genome was reported previously. These data on the At genome thus indicate that elevated recombination was a general property of allotetraploidy in cotton. PMID- 16699554 TI - Farnesol restores wild-type colony morphology to 96% of Candida albicans colony morphology variants recovered following treatment with mutagens. AB - Candida albicans is a diploid fungus that undergoes a morphological transition between budding yeast, hyphal, and pseudohyphal forms. The morphological transition is strongly correlated with virulence and is regulated in part by quorum sensing. Candida albicans produces and secretes farnesol that regulates the yeast to mycelia morphological transition. Mutants that fail to synthesize or respond to farnesol could be locked in the filamentous mode. To test this hypothesis, a collection of C. albicans mutants were isolated that have altered colony morphologies indicative of the presence of hyphal cells under environmental conditions where C. albicans normally grows only as yeasts. All mutants were characterized for their ability to respond to farnesol. Of these, 95.9% fully or partially reverted to wild-type morphology on yeast malt (YM) agar plates supplemented with farnesol. All mutants that respond to farnesol regained their hyphal morphology when restreaked on YM plates without farnesol. The observation that farnesol remedial mutants are so common (95.9%) relative to mutants that fail to respond to farnesol (4.1%) suggests that farnesol activates and (or) induces a pathway that can override many of the morphogenesis defects in these mutants. Additionally, 9 mutants chosen at random were screened for farnesol production. Two mutants failed to produce detectable levels of farnesol. PMID- 16699555 TI - A framework linkage map of perennial ryegrass based on SSR markers. AB - A moderate-density linkage map for Lolium perenne L. has been constructed based on 376 simple sequence repeat (SSR) markers. Approximately one third (124) of the SSR markers were developed from GeneThresher libraries that preferentially select genomic DNA clones from the gene-rich unmethylated portion of the genome. The remaining SSR marker loci were generated from either SSR-enriched genomic libraries (247) or ESTs (5). Forty-five percent of the GeneThresher SSRs were associated with an expressed gene. Unlike EST-derived SSR markers, GeneThresher SSRs were often associated with genes expressed at a low level, such as transcription factors. The map constructed here fulfills 2 definitions of a "framework map". Firstly, it is composed of codominant markers to ensure map transferability either within or among species. Secondly, it was constructed to achieve a level of statistical confidence in the support-for-order of marker loci. The map consists of 81 framework SSR markers spread over 7 linkage groups, the same as the haploid chromosome number. Most of the remaining 295 SSR markers have been placed into their most likely interval on the framework map. Nine RFLP markers and 1 SSR marker from another map constructed using the same pedigree were also incorporated to extend genome coverage at the terminal ends of 5 linkage groups. The final map provides a robust framework with which to conduct investigations into the genetic architecture of trait variation in this commercially important grass species. PMID- 16699556 TI - Expressed sequence tag analysis in tef (Eragrostis tef (Zucc) Trotter). AB - Tef (Eragrostis tef (Zucc.) Trotter) is the most important cereal crop in Ethiopia; however, there is very little DNA sequence information available for this species. Expressed sequence tags (ESTs) were generated from 4 cDNA libraries: seedling leaf, seedling root, and inflorescence of E. tef and seedling leaf of Eragrostis pilosa, a wild relative of E. tef. Clustering of 3603 sequences produced 530 clusters and 1890 singletons, resulting in 2420 tef unigenes. Approximately 3/4 of tef unigenes matched protein or nucleotide sequences in public databases. Annotation of unigenes associated 68% of the putative tef genes with gene ontology categories. Identification of the translated unigenes for conserved protein domains revealed 389 protein family domains (Pfam), the most frequent of which was protein kinase. A total of 170 ESTs containing simple sequence repeats (EST-SSRs) were identified and 80 EST-SSR markers were developed. In addition, 19 single-nucleotide polymorphism (SNP) and (or) insertion-deletion (indel) and 34 intron fragment length polymorphism (IFLP) markers were developed. The EST database and molecular markers generated in this study will be valuable resources for further tef genetic research. PMID- 16699557 TI - Karyological analysis of an interspecific hybrid between the dioecious Silene latifolia and the hermaphroditic Silene viscosa. AB - The genus Silene is a good model for studying evolution of the sex chromosomes, since it includes species that are hermaphroditic and dioecious, while maintain a basic chromosome number of 2n = 24. For some combinations of Silene species it is possible to construct interspecific hybrids. Here, we present a detailed karyological analysis of a hybrid between the dioecious Silene latifolia as the maternal plant and a related species, hermaphroditic Silene viscosa, used as a pollen partner. Using genomic probes (the genomic in situ hybridization (GISH) technique), we were able to clearly discriminate parental genomes and to show that they are largely separated in distinct nuclear domains. Molecular GISH and fluorescence in situ hybridization (FISH) markers document that the hybrid genome of somatic cells was strictly additive and stable, and that it had 12 chromosomes originating from each parent, including the only X chromosome of S. latifolia. Meiotic analysis revealed that, although related, respective parental chromosomes did not pair or paired only partially, which resulted in frequent chromosome abnormalities such as bridges and irregular non-disjunctions. GISH and FISH markers clearly document that the larger genome of S. latifolia and its largest chromosome component, the X chromosome, were mostly employed in chromosome lagging and misdivision. PMID- 16699558 TI - Application of comparative genomics in developing molecular markers tightly linked to the virus resistance gene Rsv4 in soybean. AB - The Rsv4 gene confers resistance to all the known strain groups of soybean mosaic virus in soybean (Glycine max (L.) Merr.). To construct a fine genetic map near Rsv4 in soybean, we employed a comparative genomics approach that used genome sequence information of the model legume Lotus japonicus. Sequences of the soybean expressed sequence tags (ESTs) AI856415 and BF070293 mapping to one side of the Rsv4 gene showed high similarity with gene sequences of the transformation competent artificial chromosome (TAC) clone LjT32P24 of Lotus. LjT32P24 is tightly linked to another sequenced TAC clone, LjT26I01, in Lotus. A new marker, AW307114A, developed from soybean EST AW307114, which is homologous to a Lotus gene within LjT26I01, was mapped to the other side of the Rsv4 gene. The identification of the microsyntenic relationship facilitated the development of additional 2 EST markers between BF070293-S and AW307114A bracketing the Rsv4 gene. Several other markers developed in this study were mapped to putative homoeologous or duplicated chromosomal regions in soybean. Alignment between the soybean maps indicated that Rsv4 is located near a local chromosomal rearrangement. This targeted comparative mapping serves to provide a foundation for marker-assisted selection and cloning of the Rsv4 gene. PMID- 16699559 TI - Quantification of the retrotransposon BARE-1 reveals the dynamic nature of the barley genome. AB - We used quantitative real-time PCR analysis to measure the copy number of the BARE-1 retrotransposon in 5 cultivars of barley (Hordeum vulgare), as well as in samples from its wild relative, Hordeum spontaneum. Two sets of PCR primers were used to amplify regions within the long terminal repeat (LTR) and the reverse transcriptase (RT) gene of BARE-1 (GenBank accession Z17327). The LTR primers detected an average of 2.148 x 105 +/- 0.012 x 105 copies per haploid genome among barley samples, whereas the RT primers detected an average of 1.588 x 104 +/- 0.085 x 104 copies. The average ratio of LTR:RT was estimated to be 13.5:1. This finding indicates that more than 7% of the barley genome is occupied by BARE 1 elements in the form of solo LTRs and another 2.6% of the genome is occupied by the full-length element. Taken together, BARE-1 sequences represent approximately 9.6% of the barley genome among the barley plants used in this study. For the above estimation, a genome size of 5.44 x 103 Mb for H. vulgare and 5.39 x 103 Mb for H. spontaneum were assumed. Our study on quantification results of the BARE-1 for a small group of barley cultivars showed that there are significant differences among cultivars in terms of BARE-1 copy number, providing further evidence that BARE-1 is active and has a major role in shaping the barley genome as a result of breeding and selection. Quantification results also showed that most of the elements (> 90%) are present as truncated copies (solo LTRs). These results show that there is a high level of recombination leading to the formation of truncated elements and a subsequent DNA loss from the genome. Taken together, our study provides a glimpse into a dynamic micro-evolutionary process that is the by-product of genome reshuffling and directional selection in barley breeding programs. PMID- 16699560 TI - Linkage and mapping analyses of the densonucleosis non-susceptible gene nsd-Z in the silkworm Bombyx mori using SSR markers. AB - In the silkworm Bombyx mori, non-susceptibility to the Zhenjiang (China) strain of the densonucleosis virus (DNV-Z) is controlled by the recessive gene nsd-Z (non-susceptible to DNV-Z), which is located on chromosome 15. Owing to a lack of crossing over in females, reciprocal backcrossed F1 (BC1) progeny were used for linkage analysis and mapping of the nsd-Z gene using silkworm strains Js and L10, which are classified as being highly susceptible and non-susceptible to DNV-Z, respectively. BC1 larvae were inoculated with the DNV-Z virus at the first instar, and DNA was extracted from the individual surviving pupae and analyzed for simple sequence repeat (SSR) markers. The nsd-Z gene was found to be linked to 7 SSR markers, as all the surviving larvae in the BC1female (F1female x L10male) showed the homozygous profile of strain L10, and the sick larvae in the BC1female (F1female x L10male) showed the heterozygous profile of Js x L10 F1 hybrids. Using a reciprocal BC1male (L101female x F1male) cross, we constructed a linkage map of 80.6 cM, with nsd-Z mapped at 30 cM and the closest SSR marker at a distance of 4.4 cM. PMID- 16699561 TI - A method to select for mutator DNA polymerase deltas in Saccharomyces cerevisiae. AB - Proofreading DNA polymerases share common short peptide motifs that bind Mg(2+) in the exonuclease active center; however, hydrolysis rates are not the same for all of the enzymes, which indicates that there are functional and likely structural differences outside of the conserved residues. Since structural information is available for only a few proofreading DNA polymerases, we developed a genetic selection method to identify mutant alleles of the POL3 gene in Saccharomyces cerevisiae, which encode DNA polymerase delta mutants that replicate DNA with reduced fidelity. The selection procedure is based on genetic methods used to identify "mutator" DNA polymerases in bacteriophage T4. New yeast DNA polymerase delta mutants were identified, but some mutants expected from studies of the phage T4 DNA polymerase were not detected. This would indicate that there may be important differences in the proofreading pathways catalyzed by the two DNA polymerases. PMID- 16699563 TI - Physical and chemical factors affecting microbial biomass and activity in contaminated subsurface riverine sediments. AB - Over 80 years of direct discharge of industrial effluents into the Mahoning River, located in northeastern Ohio, USA, has led to the accumulation of a wide variety of pollutants within its sediments. This study examined the physical and chemical parameters, including lipophilic pollutants, affecting microbial activity and biomass in subsurface (10-40 cm horizon) sediments. Microbial biomass was higher in anthropogenically contaminated sediments, and step-wise linear regression showed that approximately 82% of the variation in microbial biomass could be explained by total hexane extractable hydrocarbons, sediment particle size, and water content. There was no correlation between microbial activity and biomass. Independent variables influencing anaerobic activity were temperature and water holding capacity. The results of this study indicate that freshwater, sedimentary anaerobic microbial communities respond to a range of environmental parameters, many of which influence subsurface river sediments, and that lipophilic pollutants, when present, can cause increases in total microbial biomass. PMID- 16699564 TI - Variable adhesion and diurnal population patterns of epiphytic yeasts on creeping bentgrass. AB - Irrigation and an in vitro agitation assay were used to determine the percentage of the epiphytic yeast community (Cryptococcus, Pseudozyma, Rhodotorula, and Sporobolomyces) adhering to the phylloplane of creeping bentgrass (Agrostis palustris (Huds.) Pers.). Colony-forming units (cfu) of total epiphytic yeast populations (adherent and nonadherent cells) and of adherent populations (cells not removed by agitation) were determined by leaf washing and dilution plating. In an in vitro assay, 40.0% and 57.1% of the yeast adhered to the leaves, whereas, in initial field trials the percentage of adherent yeasts ranged from 40.0% to 71.9% of the total population. Adherent yeast cfu on leaves in the morning were significantly lower on bentgrass (8.0 x 103 to 3.1 x 104 cfu.cm(-2)) compared with total yeast cfu (1.4 x 104 to 4.7 x 104 cfu.cm(-2)) on the nonirrigated control. No differences in yeast populations were observed between irrigated and nonirrigated plots 2 h after the 0900 treatments. Yeast populations followed a diurnal pattern, with larger cfu recovered from bentgrass leaves in the morning and significantly lower populations recovered in the afternoon. At 1400 the adherent yeast were 83.1%-100% of the total yeast population recovered from the leaves. The relative adhesiveness of the epiphytic yeast community on bentgrass leaves is dynamic with nonadherent cells making up a larger percentage of the population in the mornings than the afternoons. PMID- 16699565 TI - Production of quorum-sensing-related signal molecules by epiphytic bacteria inhabiting wheat heads. AB - The production of quorum-sensing-related signal molecules (QSRMs) among culturable bacteria comprising the community on wheat heads was investigated. The taxonomic position of 186 bacterial isolates obtained from ten heads was inferred based on 16S rRNA gene sequences, and their QSRM production was determined using two bioreporter strains of N-acylhomoserine lactones. Approximately 33% of isolates produced QSRMs, though the proportion of QSRM-producing isolates on a wheat head was significantly negatively correlated with population size. Most of the producing isolates were Pantoea species, most commonly Pantoea ananatis. Furthermore, the proportion of Pantoea ananatis that produced QSRMs was significantly negatively correlated with the number of bacterial genera (community richness) on each head. Finally, community richness was positively correlated with population size. Qualitative analysis using thin-layer chromatography revealed that the QSRMs of Pantoea isolates were composed of at least two compounds. This is the first report indicating that Pantoea ananatis isolates inhabiting wheat heads are capable of producing QSRMs. QSRM production by Pantoea spp. may contribute to the predominance of this genus on wheat heads, particularly at relatively low population densities and community diversity. PMID- 16699566 TI - Model plants for studying the interaction between Methylobacterium mesophilicum and Xylella fastidiosa. AB - Over the last few years, endophytic bacterial communities associated with citrus have been studied as key components interacting with Xylella fastidiosa. In this study, we investigated the possible interaction between the citrus endophyte Methylobacterium mesophilicum SR1.6/6 and X. fastidiosa in model plants such as Catharanthus roseus (Madagaskar periwinkle) and Nicotiana clevelandii (Clevelands tobacco). The aim of this study was to establish the fate of M. mesophilicum SR1.6/6 after inoculation of C. roseus and N. clevelandii plants, using PCR-DGGE (polymerase chain reaction--denaturing gradient gel electrophoresis) and plating techniques. Shifts in the indigenous endophytic bacterial communities were observed in plants inoculated with strain SR1.6/6, using specific primers targeting alpha- and beta-Proteobacteria. Cells of strain SR1.6/6 were observed in a biofilm structure on the root and hypocotyl surfaces of in vitro seedlings inoculated with M. mesophilicum SR1.6/6. This emphasizes the importance of these tissues as main points of entrance for this organism. The results showed that C. roseus and N. clevelandii could be used as model plants to study the interaction between M. mesophilicum and X. fastidiosa. PMID- 16699567 TI - Fungal endophyte assemblages from ethnopharmaceutically important medicinal trees. AB - Endophytic fungi represent an interesting group of microorganisms associated with the healthy tissues of terrestrial plants. They represent a large reservoir of genetic diversity. Fungal endophytes were isolated from the inner bark segments of ethnopharmaceutically important medicinal tree species, namely Terminalia arjuna, Crataeva magna, Azadirachta indica, Holarrhena antidysenterica, Terminalia chebula, and Butea monosperma (11 individual trees), growing in different regions of southern India. Forty-eight fungal species were recovered from 2200 bark segments. Mitosporic fungi represented a major group (61%), with ascomycetes (21%) and sterile mycelia (18%) the next major groups. Species of Fusarium, Pestalotiopsis, Myrothecium, Trichoderma, Verticillium, and Chaetomium were frequently isolated. Exclusive fungal taxa were recovered from five of the six plant species considered for the study of endophytic fungi. Rarefaction indices for species richness indicated the highest expected number of species for bark segments were isolated from T. arjuna and A. indica (20 species each) and from C. magna (18 species). PMID- 16699569 TI - Response surface methodology for the optimization of keratinase production in culture medium containing feathers produced by Kocuria rosea. AB - A 43-fold increase in keratinase production by Kocuria rosea was achieved in batch fermentation using response surface methodology. Factorial designs were used to select the components of a culture medium that showed a significant effect on keratinase production. An orthogonal-central composite experimental design was performed, with only two (feathers and magnesium) from nine initial compounds being further analyzed by response surface methodology. An optimum keratinase production of 14 886.9 U/mg was obtained with the following medium composition (per litre): NH4Cl, 0.3 g; NaCl, 0.3 g; K2HPO4, 3.2 g; KH2PO4, 4.0 g; MgSO4.6H2O, 0.5 g; yeast extract, 0.1 g; and finely milled feathers, 30 g. The medium was shaken at 400 r/min with an incubation period of 14 h at 40 degrees C. PMID- 16699568 TI - Expression of a pertussis toxin S1 fragment by inducible promoters in oral Streptococcus and the induction of immune responses during oral colonization in mice. AB - Previous work aimed at developing a live oral vaccine expressing pertussis toxin S1 fragment on the surface of the bacterium Streptococcus gordonii elicited a lower than expected antibody response, perhaps because of low antigen expression. In this study, in-frame promoter fusions were constructed to investigate whether an increase in antigen production by the streptococcal vaccine strain results in a better antibody response. The promoters tested were (i) the Streptococcus mutans sucrose-inducible fructosyltransferase (ftf) promoter and (ii) the Bacillus subtilis/Escherichia coli chimeric tetracycline-inducible xyl/tetO promoter. Each of these two promoters was placed upstream of the spaP/s1 fusion gene to drive its expression. The constructs were introduced into S. gordonii DL1 and S. mutans 834. The inducibility of the promoters was confirmed through the determination of SpaP/S1 production via Western blottings. Induced production of SpaP/S1 was observed in S. gordonii and S. mutans with each of the promoters, but the level of expression was the highest in S. mutans, using the xyl/tetO promoter. Thus, S. mutans carrying the xyl/tetO/spaP/s1 construct (S. mutans PM14) was used in oral colonization studies in BALB/c mice. Streptococccus mutans PM14 was able to colonize the animals for the 14-week duration of experimentation. A mucosal IgA response was observed in all the treatment groups but was highest in mice receiving tetracycline induction. In the mouse model of Bordetella pertussis respiratory infection, animals colonized with S. mutans PM14 showed a decreased in B. pertussis lung colony count (P = 0.03) on day 3 compared with control mice colonized by the parent S. mutans 834. PMID- 16699570 TI - Development of AFLP-derived, functionally specific markers for environmental persistence studies of fungal strains. AB - The ability to rapidly identify and quantify a microbial strain in a complex environmental sample has widespread applications in ecology, epidemiology, and industry. In this study, we describe a rapid method to obtain functionally specific genetic markers that can be used in conjunction with standard or real time polymerase chain reaction (PCR) to determine the abundance of target fungal strains in selected environmental samples. The method involves sequencing of randomly cloned AFLP (amplified fragment length polymorphism) products from the target organism and the design of PCR primers internal to the AFLP fragments. The strain-specific markers were used to determine the fate of three industrially relevant fungi, Aspergillus niger, Aspergillus oryzae, and Chaetomium globosum, during a 4 month soil microcosm experiment. The persistence of each of the three fungal strains inoculated separately into intact soil microcosms was determined by PCR analyses of DNA directly extracted from soil. Presence and absence data based on standard PCR and quantification of the target DNA by real-time PCR showed that all three strains declined after inoculation (approximately 14-, 32-, and 4-fold for A. niger, A. oryzae, and C. globosum, respectively) but remained detectable at the end of the experiment, suggesting that these strains would survive for extended periods if released into nature. PMID- 16699571 TI - Predisposing conditions for Candida spp. carriage in the oral cavity of denture wearers and individuals with natural teeth. AB - Candida species are a normal commensal present in a large percentage of healthy individuals. Denture wearers are predisposed to the development of candidosis and to the presence of Candida spp. The presence of the yeast, even in healthy subjects, should be considered more carefully. We investigated the prevalence of Candida spp. in 112 denture wearers and 103 individuals with natural teeth, patients from the clinic of total prosthesis of the Dental School of the Federal University of Minas Gerais, Brazil, and from the School of Pharmacy and Dentistry of Alfenas, Brazil. Factors like gender, age over 60 years, low education, and xerostomia were directly associated with the presence of Candida yeasts at a significance level of 5% (p > 0.05). However, the major predisposing factor for the carrier state was wearing dentures (p = 0.001). Candida isolates were identified using morphological and biochemical profiles. Seventy-one isolates were identified as C. albicans (65.1%), 15 as C. glabrata (13.7%), 8 as C. parapsilosis (7.3%), 3 as C. krusei (2.7%), and 12 as C. tropicalis (11.0%). Susceptibility testing to fluconazole and itraconazole was also performed with the strains obtained. Both drugs showed a strong inhibition against most oral isolates. PMID- 16699572 TI - Identification of low-temperature-regulated genes in the fire blight pathogen Erwinia amylovora. AB - Genes involved in pathogenicity of several plant pathogens were shown to be induced at relatively cold temperatures. Loci from the fire blight pathogen Erwinia amylovora (Burrill) induced at 18 degrees C were identified using the miniTn5 transposon that contains the promoterless reporter gene gusA coding for beta-glucuronidase (GUS). Certain mutants (2.7%) expressed GUS predominantly at 18 degrees C on minimal medium plates, indicating that the transposon had been inserted downstream of a putatively thermoregulated promoter. Those mutants were further screened with a quantitative GUS fluorometric assay. A total of 21 mutants were selected: 19 mutants had a transposon insertion in temperature dependent genetic loci, with a 2.2- to 6.3-fold induction of gusA gene expression at 18 degrees C, and two mutants with impaired growth at 18 degrees C. Some of these genetic loci encoded (i) proteins implicated in flagella biosynthesis, biotin biosynthesis, multi-drug efflux, and type II secretion protein, and (ii) proteins of unknown function. PMID- 16699573 TI - Detection of antibiotic-related genes from bacterial biocontrol agents with polymerase chain reaction. AB - Pseudomonas chlororaphis PA23, Pseudomonas spp. strain DF41, and Bacillus amyloliquefaciens BS6 consistently inhibit infection of canola petals by Sclerotinia sclerotiorum in both greenhouse and field experiments. Bacillus thuringiensis BS8, Bacillus cereus L, and Bacillus mycoides S have shown significant inhibition against S. sclerotiorum on plate assays. The presence of antibiotic biosynthetic or self-resistance genes in these strains was investigated with polymerase chain reaction and, in one case, Southern blotting. Thirty primers were used to amplify (i) antibiotic biosythetic genes encoding phenazine-1-carboxylic acid, 2,4-diacetylphloroglucinol, pyoluteorin, and pyrrolnitrin, and (ii) the zwittermicin A self-resistance gene. Our findings revealed that the fungal antagonist P. chlororaphis PA23 contains biosynthetic genes for phenazine-1-carboxylic acid and pyrrolnitrin. Moreover, production of these compounds was confirmed by high performance liquid chromatography. Pseudomonas spp. DF41 and B. amyloliquefaciens BS6 do not appear to harbour genes for any of the antibiotics tested. Bacillus thuringiensis BS8, B. cereus L, and B. mycoides S contain the zwittermicin A self-resistance gene. This is the first report of zmaR in B. mycoides. PMID- 16699574 TI - Potential pitfalls in the quantitative molecular detection of Escherichia coli O157:H7 in environmental matrices. AB - The detection sensitivity and potential interference factors of a commonly used assay based on real-time polymerase chain reaction (PCR) for Escherichia coli O157:H7 using eae gene-specific primers were assessed. Animal wastes and soil samples were spiked with known replicate quantities of a nontoxigenic strain of E. coli O157:H7 in a viable or dead state and as unprotected DNA. The detection sensitivity and accuracy of real-time PCR for E. coli O157:H7 in animal wastes and soil is low compared to enrichment culturing. Nonviable cells and unprotected DNA were shown to produce positive results in several of the environmental samples tested, leading to potential overestimates of cell numbers due to prolonged detection of nonviable cells. This demonstrates the necessity for the specific calibration of real-time PCR assays in environmental samples. The accuracy of the eae gene-based detection method was further evaluated over time in a soil system against an activity measurement, using the bioluminescent properties of an E. coli O157:H7 Tn5luxCDABE construct. The detection of significant numbers of viable but nonculturable (VBNC) as well as nonviable and possibly physically protected cells as shown over a period of 90 days further complicates the use of real-time PCR assays for quick diagnostics in environmental samples and infers that enrichment culturing is still required for the final verification of samples found positive by real-time PCR methods. PMID- 16699575 TI - Culture conditions for the production of an acidic exopolysaccharide by the nitrogen-fixing bacterium Burkholderia tropica. AB - The endophytic diazotrophic bacterium Burkholderia tropica, strain Ppe8, produced copious amounts of exopolysaccharide (EPS) on batch growth in liquid synthetic media containing mannitol and glutamate as carbon and nitrogen sources. The effect of various aeration regimes and carbon source concentrations on EPS production was determined, as well as the effects of temperature and time of growth. The degree of aeration had a great influence on the yield of EPS, in contrast with the C:N ratio of the medium. Growth temperature also affected the EPS yield after the first 24 h of culture but seemed to be irrelevant after that. After isolation and purification, the EPS was analyzed by high-performance size exclusion chromatography and multiangle laser light scattering (HPSEC-MALLS), revealing a molecular mass of 300 kDa. The acid hydrolyzate of EPS was examined by HPLC and found to contain Glc, Rha, GlcA, and an aldobiouronic acid. The latter was found to have a GlcA and a Rha unit. Carboxy-reduced EPS contained Glc and Rha (3:2). The monosaccharide composition of the native acidic EPS was calculated as GlcA, Glc, and Rha in a molar ratio of 1:2:2. PMID- 16699576 TI - Molecular characterization of microbial communities in Canadian pulp and paper activated sludge and quantification of a novel Thiothrix eikelboomii-like bulking filament. AB - We examined the microbial community structure and quantified the levels of the filamentous bulking organism Thiothrix eikelboomii in samples of activated sludge mixed liquor suspended solids (MLSS) from Canadian pulp and paper mills. Libraries of chaperonin 60 (cpn60) gene sequences were prepared from MLSS total microbial community DNA and each was compared with cpnDB, a reference database of cpn60 sequences (http://cpndb.cbr.nrc.ca) for assignment of taxonomic identities. Sequences similar to but distinct from the type strain of T. eikelboomii AP3 (ATCC 49788T) (approximately 89% identity over 555 bp) were recovered at high frequency from a mill sample that was experiencing bulking problems at the time of sample collection, which corresponded to microscopic observations using fluorescent in situ hybridization with commercially available 16S rDNA-based probes. We enumerated this strain in five mill-derived MLSS samples using real time quantitative PCR (qPCR) and found that two samples had high levels of the bulking strain (>1012 genomes/g MLSS) and two contained lower but detectable levels of this organism. None of the mill samples contained cpn60 sequences that were identical to the type strain of T. eikelboomii. This technique shows promise for monitoring pulp and paper mill wastewater treatment systems by detecting and enumerating this strain of T. eikelboomii, which may be specific to pulp and paper mill wastewater treatment systems. PMID- 16699577 TI - Impact of concentration, temperature, and pH on inactivation of Salmonella spp. by volatile fatty acids in anaerobic digestion. AB - It is known that the presence of volatile fatty acids may play a role in the inactivation of pathogens for systems that employ an acid phase reactor. This study was conducted to investigate the influence of volatile fatty acids on the inactivation of Salmonella spp. over a range of digestion temperatures. In this study, digesters that were treating municipal wastewater treatment plant sludges were operated at temperatures that ranged from 35 to 49 degrees C and had a solids residence time of 15 days. Samples collected from the effluent of the digesters were dosed with solutions containing acetic, propionic, and butyric acids alone and in mixtures, and the dosed effluents were analyzed for Salmonella spp. over time. In the first round of testing, the digester effluents were dosed with individual organic acids and also a mixture containing all three volatile fatty acids over a range of concentrations from 750 to 6000 mg/L, and the pH of the samples was fixed at a value of 5.5. In the second round of testing, the sample sludges were spiked with a fixed amount of organic acid mixture, and the pH was varied from 4.5 to 7.5. The reduction of Salmonella spp. in digester effluents, when dosed with volatile organic acids, was found to depend on pH, temperature, the chain length of the acids, and the concentration and composition of the acids present. Increases in temperature appeared to increase the inhibitory effects of the volatile organic acids. At mesophilic temperatures, acidic pHs resulted in a greater inhibition of Salmonella spp.; whereas at higher temperatures neutral pHs were found to be more inhibitory. The results suggest that acid phase digesters that operate at elevated temperatures and low pH can achieve substantial reduction of Salmonella spp. PMID- 16699578 TI - Extracytoplasmic storage as the nickel resistance mechanism in a natural isolate of Pseudomonas putida S4. AB - Metal resistances in microbes are important to study not only to understand metal homeostasis but also to use such organisms further in environmental bioremediation. Nickel (Ni2+) is an important micronutrient, which at higher concentration becomes toxic. Many Ni2+-resistant organisms are known, which resist metal by active efflux. Pseudomonas putida S4, a natural isolate from India, is reported to show a multi-metal resistance profile. In the present study, the Ni2+-resistance mechanism in strain S4 was examined. Wild-type cells gradually accumulated Ni2+ but kept it preferentially in the periplasmic space in a bound form. In Ni2+-sensitive mutants, periplasmic storage was disturbed and more metal accumulated cytoplasmically, producing toxicity. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis analysis of periplasmic proteins revealed a band of approximately 18 kDa, which appeared only in Ni2+-exposed wild type cells, and which was absent from cells not exposed to Ni2+ as well as from Ni2+-sensitive mutants. On the basis of these observations, we propose a Ni2+ resistance mechanism in P. putida S4 based on sequestration of metal in the periplasmic space. This is the first study of sequestration-based Ni2+ resistance. PMID- 16699579 TI - Prediction and functional analysis of the replication origin of the linear plasmid pSCL2 in Streptomyces clavuligerus. AB - pSCL2 (120 kb), one of the linear plasmids found in Streptomyces clavuligerus NRRL3585, was isolated and partially sequenced. Computational analysis of the central region of pSCL2 revealed the presence of two open reading frames that appear to encode proteins highly homologous to RepL1 and RepL2, replication proteins from pSLA2-L, the large linear plasmid in Streptomyces rochei. The S. clavuligerus open reading frames were designated repC1 and repC2, encoding the proteins RepC1 (150 amino acids) and RepC2 (102 amino acids), respectively. The RepC and RepL proteins have identical translation features and very similar predicted secondary and tertiary structures. Functional analysis confirmed that RepC1 is essential for replication initiation of pSCL2, whereas RepC2 is dispensable but may play a role in copy number control. The RepC and RepL proteins do not show similarity to any other bacterial plasmid replication proteins. Three regions of DNA sequence, Box 1 (1050-850 bp), Box 2 (723-606 bp), and Box 3 (224-168 bp), located upstream of repC1, were also shown to be essential or very important for replication of pSCL2. PMID- 16699580 TI - A comparison of selected methods for measuring the virulence properties of Listeria spp. AB - The comparative ability of different methods to assess virulence of Listeria species was investigated in ten Listeria strains. All strains were initially subjected to pulsed-field gel electrophoresis analysis to determine their relatedness. Virulence characteristics were subsequently tested for by (i) determining the presence of six virulence genes by polymerase chain reaction; (ii) testing for the production of listeriolysin O, phosphatidylcholine phospholipase C, and phosphatidylinositol-specific phospholipase C; (iii) investigating the hydrophobicity of the strains; (iv) determining the strains ability to attach to, enter, and replicate within the Caco-2 cells. Variations in most of the virulence characteristics were obvious across the strains for the range of tests performed. A wide range of anomalous results among methods were apparent. In particular, the presence of virulence genes was found to be unrelated to the production of virulence-associated proteins in vitro, while virulence protein production and hydrophobicity in Listeria monocytogenes were found to be unrelated or marginally related, respectively, to the ability to invade the Caco-2 cell line. It was concluded that the methods investigated were unable to consistently and unequivocally measure the differences in the virulence properties of the strains. PMID- 16699581 TI - Tolerance and biosorption of copper and zinc by Pseudomonas putida CZ1 isolated from metal-polluted soil. AB - A strain of Pseudomonas sp. CZ1, which was isolated from the rhizosphere of Elsholtzia splendens obtained from the heavy-metal-contaminated soil in the north central region of the Zhejiang province of China, has been studied for tolerance to copper (Cu) and zinc (Zn) and its capacities for biosorption of these metals. Based on 16S ribosomal DNA sequencing, the microorganism was closely related to Pseudomonas putida. It exhibited high minimal inhibitory concentration values (about 3 mmol Cu.L-1 and 5 mmol Zn.L-1) for metals and antibiotic resistance to ampicillin but not to kanamycin. Based on the results of heavy metal toxicity screening, inhibitory concentrations in solid media were lower than those in liquid media. Moreover, it was found that the toxicity of Cu was higher than that of Zn. Pseudomonas putida CZ1 was capable of removing about 87.2% of Cu and 99.8% of Zn during the active growth cycle, with specific biosorption capacities of 24.2 and 26.0 mg x L-1, respectively. Although at low concentrations, Cu and Zn slightly damage the surface of some cells, P. putida demonstrated high capacities for biosorption of Cu and Zn. Since P. putida CZ1 could grow in the presence of significant concentrations of metals and because of its high metal uptake capacity in aerobic conditions, this bacterium may be potentially applicable in bioreactors or in situ bioremediation of heavy-metal-contaminated aqueous or soil systems. PMID- 16699582 TI - Electron transport pathways for the oxidation of endogenous substrate(s) in Acidithiobacillus ferrooxidans. AB - Oxidation of endogenous substrate(s) of Acidithiobacillus ferrooxidans with O2 or Fe3+ as electron acceptor was studied in the presence of uncouplers and electron transport inhibitors. Endogenous substrate was oxidized with a respiratory quotient (CO2 produced/O2 consumed) of 1.0, indicating its carbohydrate nature. The oxidation was inhibited by complex I inhibitors (rotenone, amytal, and piericidin A) only partially, but piericidin A inhibited the oxidation with Fe3+ nearly completely. The oxidation was stimulated by uncouplers, and the stimulated activity was more sensitive to inhibition by complex I inhibitors. HQNO (2-heptyl 4-hydroxyquinoline N-oxide) also stimulated the oxidation, and the stimulated respiration was more sensitive to KCN inhibition than uncoupler stimulated respiration. Fructose, among 20 sugars and sugar alcohols including glucose and mannose, was oxidized with a CO2/O2 ratio of 1.0 by the organism. Iron chelators in general stimulated endogenous respiration, but some of them reduced Fe3+ chemically, introducing complications. The results are discussed in view of a branched electron transport system of the organism and its possible control. PMID- 16699583 TI - Newly featured infection events in a supernodulating soybean mutant SS2-2 by Bradyrhizobium japonicum. AB - Supernodulating soybean (Glycine max L. Merr.) mutant SS2-2 and its wild-type counterpart, Sinpaldalkong 2, were examined for the microstructural events associated with nodule formation and development. SS2-2 produced a substantially higher percentage of curled root hairs than the wild type, especially at 14 days after inoculation with Bradyrhizobium japonicum. In addition, there was new evidence that in SS2-2, B. japonicum also entered through fissures created by the emerging adventitious root primordia. Early steps of nodule ontogeny were faster in SS2-2, and continued development of initiated nodules was more frequent and occurred at a higher frequency than in the wild type. These data suggest that the early expression of autoregulation is facilitated by decreasing the speed of cortical cell development, leading to the subsequent termination of less developed nodules. The nodules of SS2-2 developed into spherical nodules like those formed on the wild type. In both the wild type and supernodulating mutant, vascular bundles bifurcate from root stele and branch off in the nodule cortex to surround the central infected zone. These findings indicate that SS2-2 has complete endosymbiosis and forms completely developed nodule vascular bundles like the wild type, but that the speed of nodule ontogeny differs between the wild type and SS2-2. Thus, SS2-2 has a novel symbiotic phenotype with regard to nodule organogenesis. PMID- 16699584 TI - Fungicidal properties of two saponins from Capsicum frutescens and the relationship of structure and fungicidal activity. AB - Two steroidal saponins have been purified from cayenne pepper (Capsicum frutescens). Both have the same steroidal moiety but differ in the number of glucose moieties: the first saponin has four glucose moieties (molecular mass 1081 Da) and the second contains three glucose moieties (molecular mass 919 Da). Solubility in aqueous solution is less for the saponin containing three glucose moieties than for the one containing four glucose moieties. The larger saponin was slightly fungicidal against the nongerminated and germinating conidia of Aspergillus flavus, A. niger, A. parasiticus, A. fumigatus, Fusarium oxysporum, F. moniliforme, and F. graminearum, whereas, the second saponin (molecular mass 919 Da) was inactive against these fungi. Results indicate that the absence of one glucose molecule affects the fungicidal and aqueous solubility properties of these similar molecules. PMID- 16699585 TI - Transcriptional analysis of the Pseudomonas aeruginosa toxA regulatory gene ptxR. AB - The expression of the exotoxin A gene (toxA) in Pseudomonas aeruginosa is a complicated process that involves several regulators, including ptxR, which enhances toxA expression by 4- to 5-fold. Available evidence suggests that ptxR is expressed from two separate promoters, P1 and P2. Previous evidence indicated the presence, within the ptxR upstream region, of binding sites for several regulatory proteins, including PtxS, which negatively regulates ptxR expression. We utilized nested deletion and in vitro transcription analyses to examine the regulation of ptxR expression. The results from nested deletion analysis suggest that under aerobic conditions in iron-deficient medium, ptxR expression follows a biphasic curve that involves the P1 promoter only. Iron eliminated the second peak of ptxR expression but did not affect expression from the P2 promoter. Under microaerobic conditions, iron represses ptxR expression from subclones that carry P1 alone or P2 alone at both early and late stages of growth. Under anaerobic conditions, ptxR expression increases considerably. In addition, our results suggest that different segments of the ptxR upstream region play specific roles in ptxR expression; their deletion caused variations in the level as well as the pattern of ptxR expression. Our results also indicate that negative regulation of ptxR expression by PtxS does not occur through the PtxS binding site within the ptxR-ptxS intergenic region. In vitro transcription analysis using sigma70 reconstituted P. aeruginosa RNA polymerase produced one transcript that closely resembles T1, indicating that P1 is recognized by sigma70. RNA polymerase reconstituted with either RpoS or AlgU produced no transcripts. However, a transcript was produced by RpoH-reconstituted RNA polymerase. PMID- 16699586 TI - Foot odor due to microbial metabolism and its control. AB - To characterize foot odor, we analyzed its components by sensory tests, isolated microorganisms that produce it, and evaluated the mechanism of the occurrence of foot odor. As a result, foot odor was found to be derived from isovaleric acid, which is produced when Staphylococcus epidermidis, a resident species of the normal cutaneous microbial flora, degrades leucine present in sweat. In addition, Bacillus subtilis was detected in the plantar skin of subjects with strong foot odor, and this species was shown to be closely associated with increased foot odor. Therefore, we screened various naturally occurring substances and fragrant agents that inhibit microbial production of foot odor without disturbing the normal microbial flora of the human skin. As a result, we identified citral, citronellal, and geraniol as fragrant agents that inhibit the generation of isovaleric acid at low concentrations. PMID- 16699587 TI - Occurrence of parasporin-producing Bacillus thuringiensis in Vietnam. AB - A total of 63 Bacillus thuringiensis isolates were recovered from urban soils of Hanoi, Vietnam. Of these, 34 were identified to 12 H serogroups. None of the isolates showed larvicidal activities against three lepidopterous insects. Three isolates belonging to the two serovars, colmeri (H21) and konkukian (H34), were highly toxic to larvae of the mosquito Aedes aegypti. Parasporal inclusion proteins of four isolates exhibited cytocidal activities against HeLa cells. Immunologically, proteins of four isolates were closely related to parasporin-1 (Cry31Aa), a parasporal protein that preferentially kills human cancer cells. Haemolytic activities were associated with parasporal proteins of the three mosquitocidal isolates but not with those of the four cancer-cell-killing isolates. PCR experiments and nucleotide sequence analysis revealed that the genes of four anti-cancer isolates are closely related to the gene parasporin-1 (cry31Aa) but are dissimilar to those of the three other existing parasporins. Our results suggest that the soil of northern Vietnam is a good reservoir of parasporin-producing B. thuringiensis. PMID- 16699588 TI - Antimicrobial compounds produced by Actinomadura sp. AC104 isolated from an Algerian Saharan soil. AB - During a search for nonpolyenic antifungal antibiotics, an actinomycete designated AC104 was isolated from a Saharan soil sample by a dilution agar plating method using a chitin - vitamins B medium supplemented with rifampicin. Isolate AC104 presented the morphological and the chemical characteristics of the genus Actinomadura. On the basis of 76 physiological tests and 16S rDNA analysis, this isolate was determined to be quite different from the known species of Actinomadura. It is active against filamentous fungi and both Gram-positive and Gram-negative bacteria. The production of antibiotic substances was investigated using several culture media. The highest antimicrobial activities were obtained on ISP2 medium. The benzenic extract contained five bioactive spots detected on thin layer chromatography plates. Among these antibiotics, a complex called 104A, which showed the more interesting antifungal activity, was selected and purified by reverse-phase high-pressure liquid chromatography. This complex is composed of four compounds. Ultraviolet-visible, infrared, mass, and 1H nuclear magnetic resonance spectroscopy studies showed that these molecules contain an aromatic ring substituted by aliphatic chains. These compounds differ from the known antibiotics produced by Actinomadura species. PMID- 16699589 TI - Effects of amoebae on the growth of microbes isolated from moisture-damaged buildings. AB - Dampness, moisture, and mold in buildings are associated with adverse health outcomes. In addition to fungi and bacteria, amoebae have been found in moisture damaged building materials. Amoebae and a growing list of bacteria have been shown to have mutual effects on each other's growth, but the interactions between amoebae and microbes common in moisture-damaged buildings have not been reported. We co-cultivated the amoeba Acanthamoeba polyphaga with bacteria and fungi isolated from moisture-damaged buildings in laboratory conditions for up to 28 days. The microbes selected were the bacteria Streptomyces californicus, Bacillus cereus, and Pseudomonas fluorescens, and the fungi Stachybotrys chartarum, Aspergillus versicolor, and Penicillium spinulosum. Fungi and bacteria generally benefited from the presence of the amoebae, whereas the growth of amoebae was hindered by Streptomyces californicus, Stachybotrys chartarum, and Bacillus cereus. Pseudomonas fluorescens slightly enhanced amoebae viability. Amoebae were indifferent to the presence of Aspergillus versicolor and Penicillium spinulosum. Thus, our results show that amoebae can alter the survival and growth of some microbes in moisture-damaged buildings. PMID- 16699590 TI - Fhua and HgbA, outer membrane proteins of Actinobacillus pleuropneumoniae: their role as virulence determinants. AB - For the recently described serotype 15 of biotype I and serotypes 13 and 14 of biotype II of Actinobacillus pleuropneumoniae, fhuA and hgbA were detected by polymerase chain reaction and DNA sequencing. To determine the substrate specificity of the iron receptors FhuA and HgbA and to study their role in the virulence of A. pleuropneumoniae, we used two isogenic A. pleuropneumoniae serotype 1 deletion mutants of fhuA and hgbA. Different sources of iron and siderophores were tested in growth promotion assays. FhuA and HgbA are specific for their ligands ferrichrome and hemoglobin, respectively. The virulence of the two deletion mutant strains was evaluated in experimental infections using specific pathogen-free piglets. While the fhuA mutant (DG02) was as highly virulent as the parental strain S4074, the virulence of the hgbA mutant (DeltahgbA) was reduced. Our data indicate that both FhuA and HgbA are conserved among all serotypes and biotypes of A. pleuropneumoniae and that HgbA, the receptor for porcine hemoglobin, may play a role in virulence. PMID- 16699591 TI - Novel route to 5-position vinyl derivatives of thiolactomycin: Olefination vs. deformylation. AB - Vinyl and diene derivatives of thiolactomycin have been prepared via Horner Wadsworth-Emmons olefination from protected 5-formyl-3,5-dimethylthiotetronic acid. Several 4-position protecting groups and a variety of phosphonates were evaluated, with MOM protection and beta-ketophosphonates yielding the highest ratio of desired product to deformylated product. PMID- 16699592 TI - Risk alleles of USF1 gene predict cardiovascular disease of women in two prospective studies. AB - Upstream transcription factor 1 (USF1) is a ubiquitously expressed transcription factor controlling several critical genes in lipid and glucose metabolism. Of some 40 genes regulated by USF1, several are involved in the molecular pathogenesis of cardiovascular disease (CVD). Although the USF1 gene has been shown to have a critical role in the etiology of familial combined hyperlipidemia, which predisposes to early CVD, the gene's potential role as a risk factor for CVD events at the population level has not been established. Here we report the results from a prospective genetic-epidemiological study of the association between the USF1 variants, CVD, and mortality in two large Finnish cohorts. Haplotype-tagging single nucleotide polymorphisms exposing all common allelic variants of USF1 were genotyped in a prospective case-cohort design with two distinct cohorts followed up during 1992-2001 and 1997-2003. The total number of follow-up years was 112,435 in 14,140 individuals, of which 2,225 were selected for genotyping based on the case-cohort study strategy. After adjustment for conventional risk factors, we observed an association of USF1 with CVD and mortality among females. In combined analysis of the two cohorts, female carriers of a USF1 risk haplotype had a 2-fold risk of a CVD event (hazard ratio [HR] 2.02; 95% confidence interval [CI] 1.16-3.53; p = 0.01) and an increased risk of all-cause mortality (HR 2.52; 95% CI 1.46-4.35; p = 0.0009). A putative protective haplotype of USF1 was also identified. Our study shows how a gene identified in exceptional families proves to be important also at the population level, implying that allelic variants of USF1 significantly influence the prospective risk of CVD and even all-cause mortality in females. PMID- 16699594 TI - Inferring loss-of-heterozygosity from unpaired tumors using high-density oligonucleotide SNP arrays. AB - Loss of heterozygosity (LOH) of chromosomal regions bearing tumor suppressors is a key event in the evolution of epithelial and mesenchymal tumors. Identification of these regions usually relies on genotyping tumor and counterpart normal DNA and noting regions where heterozygous alleles in the normal DNA become homozygous in the tumor. However, paired normal samples for tumors and cell lines are often not available. With the advent of oligonucleotide arrays that simultaneously assay thousands of single-nucleotide polymorphism (SNP) markers, genotyping can now be done at high enough resolution to allow identification of LOH events by the absence of heterozygous loci, without comparison to normal controls. Here we describe a hidden Markov model-based method to identify LOH from unpaired tumor samples, taking into account SNP intermarker distances, SNP-specific heterozygosity rates, and the haplotype structure of the human genome. When we applied the method to data genotyped on 100 K arrays, we correctly identified 99% of SNP markers as either retention or loss. We also correctly identified 81% of the regions of LOH, including 98% of regions greater than 3 megabases. By integrating copy number analysis into the method, we were able to distinguish LOH from allelic imbalance. Application of this method to data from a set of prostate samples without paired normals identified known regions of prevalent LOH. We have developed a method for analyzing high-density oligonucleotide SNP array data to accurately identify of regions of LOH and retention in tumors without the need for paired normal samples. PMID- 16699595 TI - Differential repression of alternative transcripts: a screen for miRNA targets. AB - Alternative polyadenylation sites produce transcript isoforms with 3' untranslated regions (UTRs) of different lengths. If a microRNA (miRNA) target is present in the UTR, then only those target-containing isoforms should be sensitive to control by a cognate miRNA. We carried out a systematic examination of 3' UTRs containing multiple poly(A) sites and putative miRNA targets. Based on expressed sequence tag (EST) counts and EST library information, we observed that levels of isoforms containing targets for miR-1 or miR-124, two miRNAs causing downregulation of transcript levels, were reduced in tissues expressing the corresponding miRNA. This analysis was repeated for all conserved 7-mers in 3' UTRs, resulting in a selection of 312 motifs. We show that this set is significantly enriched in known miRNA targets and mRNA-destabilizing elements, which validates our initial hypothesis. We scanned the human genome for possible cognate miRNAs and identified phylogenetically conserved precursors matching our motifs. This analysis can help identify target-miRNA couples that went undetected in previous screens, but it may also reveal targets for other types of regulatory factors. PMID- 16699593 TI - The leukocyte receptor complex in chicken is characterized by massive expansion and diversification of immunoglobulin-like Loci. AB - The innate and adaptive immune systems of vertebrates possess complementary, but intertwined functions within immune responses. Receptors of the mammalian innate immune system play an essential role in the detection of infected or transformed cells and are vital for the initiation and regulation of a full adaptive immune response. The genes for several of these receptors are clustered within the leukocyte receptor complex (LRC). The purpose of this study was to carry out a detailed analysis of the chicken (Gallus gallus domesticus) LRC. Bacterial artificial chromosomes containing genes related to mammalian leukocyte immunoglobulin-like receptors were identified in a chicken genomic library and shown to map to a single microchromosome. Sequencing revealed 103 chicken immunoglobulin-like receptor (CHIR) loci (22 inhibitory, 25 activating, 15 bifunctional, and 41 pseudogenes). A very complex splicing pattern was found using transcript analyses and seven hypervariable regions were detected in the external CHIR domains. Phylogenetic and genomic analysis showed that CHIR genes evolved mainly by block duplications from an ancestral inhibitory receptor locus, with transformation into activating receptors occurring more than once. Evolutionary selection pressure has led not only to an exceptional expansion of the CHIR cluster but also to a dramatic diversification of CHIR loci and haplotypes. This indicates that CHIRs have the potential to complement the adaptive immune system in fighting pathogens. PMID- 16699596 TI - Spatial regulation and the rate of signal transduction activation. AB - Of the many important signaling events that take place on the surface of a mammalian cell, activation of signal transduction pathways via interactions of cell surface receptors is one of the most important. Evidence suggests that cell surface proteins are not as freely diffusible as implied by the classic fluid mosaic model and that their confinement to membrane domains is regulated. It is unknown whether these dynamic localization mechanisms function to enhance signal transduction activation rate or to minimize cross talk among pathways that share common intermediates. To determine which of these two possibilities is more likely, we derive an explicit equation for the rate at which cell surface membrane proteins interact based on a Brownian motion model in the presence of endocytosis and exocytosis. We find that in the absence of any diffusion constraints, cell surface protein interaction rate is extremely high relative to cytoplasmic protein interaction rate even in a large mammalian cell with a receptor abundance of a mere two hundred molecules. Since a larger number of downstream signaling events needs to take place, each occurring at a much slower rate than the initial activation via association of cell surface proteins, we conclude that the role of co-localization is most likely that of cross-talk reduction rather than coupling efficiency enhancement. PMID- 16699597 TI - Dynamic changes in subgraph preference profiles of crucial transcription factors. AB - Transcription factors with a large number of target genes--transcription hub(s), or THub(s)--are usually crucial components of the regulatory system of a cell, and the different patterns through which they transfer the transcriptional signal to downstream cascades are of great interest. By profiling normalized abundances (A(N)) of basic regulatory patterns of individual THubs in the yeast Saccharomyces cerevisiae transcriptional regulation network under five different cellular states and environmental conditions, we have investigated their preferences for different basic regulatory patterns. Subgraph-normalized abundances downstream of individual THubs often differ significantly from that of the network as a whole, and conversely, certain over-represented subgraphs are not preferred by any THub. The THub preferences changed substantially when the cellular or environmental conditions changed. This switching of regulatory pattern preferences suggests that a change in conditions does not only elicit a change in response by the regulatory network, but also a change in the mechanisms by which the response is mediated. The THub subgraph preference profile thus provides a novel tool for description of the structure and organization between the large-scale exponents and local regulatory patterns. PMID- 16699598 TI - HIV-1 Vpu promotes release and prevents endocytosis of nascent retrovirus particles from the plasma membrane. AB - The human immunodeficiency virus (HIV) type-1 viral protein U (Vpu) protein enhances the release of diverse retroviruses from human, but not monkey, cells and is thought to do so by ablating a dominant restriction to particle release. Here, we determined how Vpu expression affects the subcellular distribution of HIV-1 and murine leukemia virus (MLV) Gag proteins in human cells where Vpu is, or is not, required for efficient particle release. In HeLa cells, where Vpu enhances HIV-1 and MLV release approximately 10-fold, concentrations of HIV-1 Gag and MLV Gag fused to cyan fluorescent protein (CFP) were initially detected at the plasma membrane, but then accumulated over time in early and late endosomes. Endosomal accumulation of Gag-CFP was prevented by Vpu expression and, importantly, inhibition of plasma membrane to early endosome transport by dominant negative mutants of Rab5a, dynamin, and EPS-15. Additionally, accumulation of both HIV and MLV Gag in endosomes required a functional late budding domain. In human HOS cells, where HIV-1 and MLV release was efficient even in the absence of Vpu, Gag proteins were localized predominantly at the plasma membrane, irrespective of Vpu expression or manipulation of endocytic transport. While these data indicated that Vpu inhibits nascent virion endocytosis, Vpu did not affect transferrin endocytosis. Moreover, inhibition of endocytosis did not restore Vpu-defective HIV-1 release in HeLa cells, but instead resulted in accumulation of mature virions that could be released from the cell surface by protease treatment. Thus, these findings suggest that a specific activity that is present in HeLa cells, but not in HOS cells, and is counteracted by Vpu, traps assembled retrovirus particles at the cell surface. This entrapment leads to subsequent endocytosis by a Rab5a- and clathrin dependent mechanism and intracellular sequestration of virions in endosomes. PMID- 16699600 TI - A rant against jargon and neologisms. PMID- 16699599 TI - Human retroviral host restriction factors APOBEC3G and APOBEC3F localize to mRNA processing bodies. AB - APOBEC3G is an antiviral host factor capable of inhibiting the replication of both exogenous and endogenous retroviruses as well as hepatitis B, a DNA virus that replicates through an RNA intermediate. To gain insight into the mechanism whereby APOBEC3G restricts retroviral replication, we investigated the subcellular localization of the protein. Herein, we report that APOBEC3G localizes to mRNA processing (P) bodies, cytoplasmic compartments involved in the degradation and storage of nontranslating mRNAs. Biochemical analysis revealed that APOBEC3G localizes to a ribonucleoprotein complex with other P-body proteins which have established roles in cap-dependent translation (eIF4E and eIF4E-T), translation suppression (RCK/p54), RNA interference-mediated post-transcriptional gene silencing (AGO2), and decapping of mRNA (DCP2). Similar analysis with other APOBEC3 family members revealed a potential link between the localization of APOBEC3G and APOBEC3F to a common ribonucleoprotein complex and P-bodies with potent anti-HIV-1 activity. In addition, we present evidence suggesting that an important role for HIV-1 Vif, which subverts both APOBEC3G and APOBEC3F antiviral function by inducing their degradation, could be to selectively remove these proteins from and/or restrict their localization to P-bodies. Taken together, the results of this study reveal a novel link between innate immunity against retroviruses and P-bodies suggesting that APOBEC3G and APOBEC3F could function in the context of P-bodies to restrict HIV-1 replication. PMID- 16699603 TI - Interrelations between psychiatric symptoms and drug-induced movement disorder. AB - After 30 years of clinical research into drug-induced movement disorder (DIMD), we are still facing unresolved issues regarding the interrelations between psychiatric symptoms and DIMD. Recently, I proposed a new classification of DIMD that includes abnormal movements previously labelled extrapyramidal symptoms. DIMD caused by psychotropic drugs is still confused with psychiatric symptoms treated by the same drugs. The results from 2 international multicentre trials, the InterSePT and the Ris-Consta Studies, conducted in the era of both typical and atypical antipsychotic agents, which included over 3,000 patients with schizophrenia and schizoaffective disorder worldwide, still showed a high, but decreasing, incidence of pretreatment DIMD, which varied from 57.5% (1998-1999) to 47.4% (1999-2000), and a decreasing incidence of tardive dyskinesia, which varied from 12% (1998-1999) to 10.2% (1999-2000), reflecting the greater use of atypical antipsychotic drugs. Furthermore, in both studies, psychiatric symptoms as measured by the Positive and Negative Symptom Scale (PANSS) were significantly correlated with DIMD and DIMD subtypes, thus suggesting the need for additional measurement instruments in schizophrenia and related psychoses. PMID- 16699602 TI - The search for new off-label indications for antidepressant, antianxiety, antipsychotic and anticonvulsant drugs. AB - Most drugs are prescribed for several illnesses, but it took several years for psychotropic drugs to have multiple clinical indications. Our search for serotonergic drugs in affective illnesses and related disorders led to new off label indications for fluoxetine, sertraline, tryptophan, clonazepam, alprazolam, tomoxetine, buproprion, duloxetine, risperidone and gabapentin. Various clinical trial designs were used for these proof-of-concept studies. Novel therapeutic uses of benzodiazepines, such as in panic disorder and mania, were found with the introduction of 2 high-potency benzodiazepines, clonazepam and alprazolam, which were thought to have serotonergic properties. Our initial clinical trials of fluoxetine and sertraline led to their approved indications in the treatment of obsessive-compulsive disorder, and our trials of gabapentin led to new indications in anxiety disorders (generalized anxiety, panic attack and social phobia) and sleep disorders (insomnia). PMID- 16699604 TI - "Missing links" in borderline personality disorder: loss of neural synchrony relates to lack of emotion regulation and impulse control. AB - OBJECTIVE: Symptoms of borderline personality disorder (BPD) may reflect distinct breakdowns in the integration of posterior and frontal brain networks. We used a high temporal resolution measure (40-Hz gamma phase synchrony) of brain activity to examine the connectivity of brain function in BPD. METHODS: Unmedicated patients with BPD (n = 15) and age-and sex-matched healthy control subjects (n = 15) undertook a task requiring discrimination of salient from background tones. In response to salient stimuli, the magnitude and latency of peak gamma phase synchrony for early (0-150 ms post stimulus) and late (250-500 ms post stimulus) phases were calculated for frontal and posterior regions and for left and right hemispheres. We recorded skin conductance responses (SCRs) and reaction time (RT) simultaneously to examine the contribution of arousal and performance. RESULTS: Compared with controls, patients with BPD had a significant delay in early posterior gamma synchrony and a reduction in right hemisphere late gamma synchrony in response to salient stimuli. Both SCR onset and RT were also delayed in BPD, but independently from differences in synchrony. The delay in posterior synchrony was associated with cognitive symptoms, and reduced right hemisphere synchrony was associated with impulsivity. CONCLUSIONS: These findings suggest that distinct impairments in the functional connectivity of neural systems for orienting to salient input underlie core dimensions of cognitive disturbance and poor impulse control in BPD. PMID- 16699605 TI - Downregulation in components of the mitochondrial electron transport chain in the postmortem frontal cortex of subjects with bipolar disorder. AB - OBJECTIVE: Many studies indicate a genetic predisposition to bipolar disorder (BD) and suggest that a number of abnormal genes are involved in its development. In this study, we used DNA microarray technology to analyze gene-expression profiles in the postmortem frontal cortex of subjects with BD. METHODS: Microarray hybridization was performed using human 19K microarray with universal human reference RNA in each hybridization. The reference cDNA was labelled with Cy3 and experimental cDNA, with Cy5. Glass array slides were cohybridized with equal amounts of mixed reference and experimental cDNA. Selected gene targets were further verified using real-time polymerase chain reaction (PCR). RESULTS: We found that 831 genes were differentially expressed in subjects with BD, including a number of genes in the mitochondrial electron transport chain (ETC), phosphatidylinositol-signalling system and glycolysis/ gluconeogenesis. Eight genes coding for the components of the mitochondrial ETC were identified along with 15 others related to mitochondrial function. Downregulation of NADH ubiquinone oxidoreductase 20-kd subunit (ETC complex I), cytochrome c oxidase polypeptide Vic (ETC complex IV) and ATP synthase lipid-binding protein (ETC complex V) were further verified by real-time PCR. We also found that the expression of the NADH-ubiquinone oxidoreductase 20-kd subunit was increased in subjects with BD who were receiving mood-stabilizing treatment with lithium at the time of death, when compared with subjects with BD who were not being treated with lithium. CONCLUSIONS: Because the mitochondrial ETC is a major source for the generation of reactive oxygen species, these findings suggest that oxidative damage may play an important role in the pathophysiology of BD and that neuroprotection against this damage may be involved in the effect of lithium treatment. PMID- 16699601 TI - Schizophrenia: an integrative approach to modelling a complex disorder. AB - The discovery of candidate susceptibility genes for schizophrenia and the generation of mice lacking proteins that reproduce biochemical processes that are disrupted in this mental illness offer unprecedented opportunities for improved modelling of this complex disorder. Several lines of evidence indicate that obstetrical complications, as well as fetal or neonatal exposure to viral infection, are predisposing events for some forms of schizophrenia. These environmental events can be modelled in animals, resulting in some of the characteristic features of schizophrenia; however, animal models have yet to be developed that encompass both environmental and genetic aspects of this mental illness. A large number of candidate schizophrenia susceptibility genes have been identified that encode proteins implicated in the regulation of synaptic plasticity, neurotransmission, neuronal migration, cell adherence, signal transduction, energy metabolism and neurite outgrowth. In support of the importance of these processes in schizophrenia, mice that have reduced levels or completely lack proteins that control glutamatergic neurotransmission, neuronal migration, cell adherence, signal transduction, neurite outgrowth and synaptic plasticity display many features reminiscent of schizophrenia. In the present review, we discuss strategies for modelling schizophrenia that involve treating mice that bear these mutations in a variety of ways to better model both environmental and genetic factors responsible for this complex mental illness according to a "two-hit hypothesis." Because rodents are able to perform complex cognitive tasks using odour but not visual or auditory cues, we hypothesize that olfactory-based tests of cognitive performance should be used to search for novel therapeutics that ameliorate the cognitive deficits that are a feature of this devastating mental disorder. PMID- 16699606 TI - The rational use of medications in acute psychotic presentations -- the case for less is more. PMID- 16699607 TI - [A life and a complete labor]. PMID- 16699609 TI - [Professor Jose Fernandes Pontes and social psychosomatic Medicine]. PMID- 16699611 TI - [Role of the genetic polymorphism of p53 (codon 72) gene in colorectal cancer]. AB - BACKGROUND: Polymorphisms are genetic variations that can occur in sequences of codons, leading to defective proteins. p53 is the most commonly gene affected in human cancer. The polymorphism of this gene occurs by a substitution of a base in codon 72 and may increase the risk of cancer. AIM: To investigate the possible association between p53 arginine/72 proline polymorphism and susceptibility to colorectal cancer. PATIENTS AND METHODS: This polymorphism was studied by polymerization chain reaction using specific primers in 100 patients with colorectal cancer paired by sex and age to 100 patients without cancer. Alcohol and tobacco used by all the patients and clinical aspects as stage, grade of differentiation and recurrence in the case group was compared with the genotype analyzed. RESULTS: The frequency of homozygosis for arginine was 56% in the cancer group and 58% in the control group. No significant difference was observed among both groups. This genotype was more frequent in colorectal cancer patients stage IV than in stage I (80% versus 14%). There was no significant difference between genotypes and alcohol, tobacco, grade of differentiation or recurrence. CONCLUSION: Homozygosity for arginine was the most prevalent genotype in both groups. The frequency of codon 72 proline/arginine p53 gene polymorphism was not correlated with a higher risk of colorectal cancer. Arginine/arginine genotype was more prevalent in advanced cancer patients (stage IV). PMID- 16699610 TI - [p53 in clinical contexts: yes or not?]. PMID- 16699612 TI - Expression of the p53 protein and clinical and pathologic correlation in adenocarcinoma of the rectum. AB - BACKGROUND: p53 is a tumor suppressor gene that has been implicated in the pathogenesis of a wide range of tumors, including colorectal cancer. AIM: To evaluate the prognostic significance of p53 protein expression, and to correlate with clinical and pathologic variables as well as survival. PATIENTS AND METHODS: Immunohistochemical staining for p53 protein was performed in a series of 83 patients with rectal cancer with a follow-up of at least 5 years. RESULTS: Expression of nuclear protein was positive in 36 (43.4%) of the tumors. We found association between positive p53 and death, local recurrence, distant metastasis and overall recurrence and we also found association between p53 and death, local recurrence, metastasis and overall recurrence when analysis was adjusted to Astler-Coller stage and grade of differentiation. CONCLUSION: p53 immunoexpression has shown to be an independent prognostic factor in these series. PMID- 16699613 TI - Comparison between endoscopic brush cytology performed before and after biliary stricture dilation for cancer detection. AB - BACKGROUND: Confirmation of malignancy within biliary strictures is endoscopically challenging. Dilation of strictures has been reported to enhance cytological diagnosis. AIM: To compare brush cytology results before and after biliary stricture dilation. PATIENTS AND METHODS: Patients with extra-hepatic biliary stricture at endoscopic retrograde cholangiopancreatography were included in the study. Brushing was performed before and immediately after dilation using a 10 Fr dilating catheter. Cytology samples were classified as: negative for malignancy, presence of atypical cells, insufficient material, suspicious for malignancy or positive for malignancy. Final diagnosis was established by surgery, biopsy or follow-up. RESULTS: Biliary brush cytology was performed in 50 patients, with an overall sensitivity of 40% and 27.5%, before and after dilation, respectively. The combination of results increased cancer detection rate to 45%. There were 5/50 (10%) minor complications and one death related to the procedure. CONCLUSIONS: Brush cytology performed before biliary stricture dilation has a similar cancer detection rate to that following dilation, although the combination of results enhances sensitivity. PMID- 16699614 TI - Hepatocellular carcinoma in patients undergoing orthotopic liver transplantation: radiological findings with anatomopathological correlation in Brazil. AB - BACKGROUND: Hepatocellular carcinoma is one of the most common malignant tumors worldwide. Imaging techniques, specially computed tomography and ultrasound, are among the most useful diagnostic tools, although the accuracy of these methods may have a significant variability. AIMS: To determine the prevalence of hepatocellular carcinoma in cirrhotic patients undergoing orthotopic liver transplantation at "Santa Casa de Misericordia" of Porto Alegre, RS, Brazil; to estimate the sensitivity of computed tomography and ultrasound in pretransplantation detection of hepatocellular carcinoma in this population; to correlate the radiological characteristics with anatomopathological findings. MATERIALS AND METHODS: Retrospective prevalence study. POPULATION: adult, cirrhotic patients undergoing orthotopic liver transplantation from January 1990 to July 2003. Among the 292 transplanted patients, 31 cases of hepatocellular carcinoma were diagnosed, of which 29 were included in the study. Tumor characteristics in both ultrasound and computed tomography were compared to those observed in anatomopathological examination. RESULTS: Prevalence of hepatitis C virus infection among patients with diagnosis of hepatocellular carcinoma was 93.5%, and the prevalence of hepatocellular carcinoma among transplanted patients was 10.6%. The overall sensitivity of the imaging techniques was 70.3% for computed tomography and 72% for ultrasound. CONCLUSION: The prevalence of hepatocellular carcinoma at our institution, as well as the sensitivity of both ultrasound and computed tomography to detect such tumors at pretransplantation screening were similar to those found by other authors, while the prevalence of hepatitis C virus infection, the most common etiological agent for liver disease in our patients, is one of the highest ever reported in literature. Factors influencing hepatocellular carcinoma detection rates were: time from examination to liver transplantation; acquisition of computed tomography images during arterial phase; lesion size. Arterial phase proved to be the most useful part of computed tomography examination in this study. PMID- 16699615 TI - Curative resection plus adjuvant chemotherapy for early stage primary gastric non Hodgkin's lymphoma: a retrospective study with emphasis on prognostic factors and treatment outcome. AB - BACKGROUND: There is controversy regarding the optimal therapy for primary non Hodgkin gastric lymphoma with some authors defending surgical extirpation either alone or in association with radiotherapy and or chemotherapy, especially in relation to the earlier stages of the disease. AIM: To analyze the clinical pathological features and the results of management approaches for patients with primary early-stage non-Hodgkin's lymphoma of the stomach operated in Surgical Gastroenterology Department, "Hospital do Servidor Publico Estadual", Sao Paulo, SP, Brazil. The literature is reviewed to highlight the aspects of diagnosis, prognostic factors and role of the various treatment regimens. METHOD: Sixteen patients with primary early-stage gastric lymphoma underwent curative surgical treatment. The variables analyzed were age, sex, location, size, type of surgery, number of lesions, depth of invasion, histological type in accordance with Kiel's classification, involvement of lymph nodes, Ann Arbor stage classification modified by Musshoff and Schmidt-Vollmer, histological grade, margins, adjuvant therapy, clinical course and survival. RESULTS: Ten patients (62.5%) underwent subtotal gastrectomy and six (37.5%) underwent total gastrectomy. The majority (9/56.2%) of the lesions were located in the antrum. Single lesions (10/62.5%) were more frequent than multiple lesions (6/37.5%). Thirteen patients (81.2%) were classified as stage IE and three (18.7%) as stage IIE1. Primary gastric lymphoma classified histologically as low or high grade was presented by 10 (62.5%) and 6 (37.5%) patients, respectively. The most frequent histological types were the lymphoplasmocytic cytoid (4/25.0%) and centroblastic (4/25.0%). Ten patients (62.5%) received adjuvant treatment (chemotherapy and/or radiotherapy). Nine patients (56.2%), all in stage IE, reached a survival greater than 5 years and of these eight (50.0.%) had received adjuvant therapy. Two (12.5%) patients with stage IIE1 presented peritoneal relapse and died 3.0 years and 3.5 years after their respective operations. The mean overall survival was 42.5 months. CONCLUSIONS: Among the patients with primary early-stage gastric lymphoma (IE and IIE1), the gastric resection enabled an accurate clinicopathological staging, in addition to obtaining sufficient material for histopathological study and extirpation of the lesion. Furthermore, for patients with stage IE disease, the gastric resection combined with adjuvant therapy was associated with a greater than 5-year survival. Until prospective randomized studies are realized in order to evaluate the real efficacy of the different types of treatment for primary early-stage gastric lymphoma, management approaches should be individually tailored. PMID- 16699616 TI - [Relationship between the size of hiatal hernia and esophageal acid exposure time in erosive and non-erosive reflux disease]. AB - BACKGROUND: In the last few years studies have demonstrated that hiatal hernias have an important role in the pathogenesis of reflux disease, promoting reflux by many different mechanisms, emphasizing that the larger the hiatal hernia, the higher the reflux intensity and erosive esophagitis prevalence. AIM: To correlate the size of hiatal hernias (small or large) with reflux intensity (measured by pH monitoring parameters) in patients with non-erosive and erosive reflux disease. PATIENTS AND METHODS: We reviewed pH monitoring from patients with typical reflux symptoms (heartburn) previously submitted to upper endoscopy. Reflux intensity was measured by the % of total time of pH < 4 (%TT) and by % of time of pH < 4 in upright (%UT) and supine (%ST) positions. Patients were classified as non-erosive reflux disease if no erosive esophagitis was found in endoscopy and pH monitoring was abnormal and as erosive reflux disease if they had erosive esophagitis. Hiatal hernias were classified as small if their size ranged from 2 to 4 cm and large if > or = 5 cm. RESULTS: A total of 192 patients were included, being 115 in erosive reflux disease group and 77 in non-erosive reflux disease group. In erosive reflux disease patients, there were 94 (81%) with small hiatal hernias and 21 (19%) with large ones and in non-erosive reflux disease patients there were 66 (85%) with small and 11(15%) with large hiatal hernias. In erosive reflux disease group, the %TT, %UT and %ST were of 13.1 +/- 7.1; 13.4 +/- 7.4 and 12.3 +/- 11.5 in small hiatal hernias and 20.2 +/- 12.3; 17.8 +/- 14.1 and 20.7 +/- 14.1 in large hiatal hernias, respectively (P < 0.05 for %TT and %TS). In non erosive reflux disease patients, %TT, %UT and %ST were 9.6 +/- 4.8; 10.8 +/- 6.8 and 8.6 +/- 7.3 in small hiatal hernias and of 14.6 +/- 13.3; 11.2 +/- 7.5 and 18.1 +/- 21.0 in large hiatal hernias respectively (P < 0.05 for %TT and %TS). CONCLUSION: Large hiatal hernias increase acid exposure time only in supine position in erosive esophagitis patients and in non-erosive patients. PMID- 16699617 TI - [13C-methacetin breath test in chronic hepatitis C liver disease]. AB - BACKGROUND: The 13C-methacetin breath test is a non-invasive method to evaluate hepatic microsomal function that allows a quantitative assessment of the functional hepatic mass. AIM: To evaluate the clinical usefulness of the 13C methacetin breath test in patients with hepatitis C chronic liver disease. PATIENTS AND METHODS: Seventy eight patients with chronic hepatitis C and 13 matched healthy controls were studied. HCV patients were classified as having chronic hepatitis (n = 51), cirrhosis (n = 27), being seven with decompensated disease (presence of ascite, jaundice and/or encephalopathy). HbsAg/HIV co infected patients, chronic alcohol drinker, having other chronic diseases and those using drugs that could interfere with hepatic cytochrome P450, were excluded. The disease stage and activity in biopsy fragments were determined according the Brazilian Society of Hepatology criteria. Breath test was performed with 75 mg of 13C-methacetin, and the 13CO2 in the expired air was measured through a nondispersive infra red spectrometry. The delta over baseline, and the cumulative recovery of 13CO2 at 40 (13C-methacetin breath test 40 min) and 120 minutes (13C-methacetin breath test 120 min) were calculated. RESULTS: 13C methacetin breath test parameters correlate only with hepatic staging but not with necroinflammatory (activity) parameters, being the best correlation found between hepatic staging and the 13C-methacetin breath test 120 minutes. The mean values for 13C-methacetin breath test 120 min was significantly reduced in the cirrhotic groups (19.2 +/- 7.1% for compensated and 14.7 +/- 4.0% for decompensated cirrhotics) than in control (29.9 +/- 4.5%) and chronic hepatitis (27.8 +/- 6.1%) groups. The best diagnostic accuracy for the diagnosis of cirrhosis among HCV patients was found for 13C-methacetin breath test 120 min with 81% of sensibility and 77% of specificity. CONCLUSION: 13C-methacetin breath test is correlated with structural changes in HCV-related chronic hepatic diseases and the cumulative recovery at 120 minutes is a sensitive parameter to identify the presence of hepatic cirrhosis in these patients. PMID- 16699618 TI - Clinical and biochemical features of autoimmune hepatitis in 36 pediatric patients. AB - BACKGROUND: Few studies on autoimmune hepatitis have enrolled non-Caucasian groups. AIMS: To evaluate Brazilian children with type 1 and 2 autoimmune hepatitis regarding outcome and clinical and biochemical parameters. PATIENTS AND METHODS: Thirty-six patients were submitted to a protocol that evaluated the clinical history, physical and biochemical data, and the course of the disease. Twenty-four children had type 1 autoimmune hepatitis, seven had type 2 and five had unclassified autoimmune hepatitis. Most patients were females (77%), with a median age at diagnosis of 11 years, and the median duration of symptoms was 5.5 and 8 months for types 1 and 2, respectively. Jaundice and choluria were the most common clinical manifestations. RESULTS: Treatment with azathioprine and prednisone was successful in patients with type 1 and 2 autoimmune hepatitis. AST and ALT decreased after 4 to 8 weeks of treatment compared to pretreatment levels in type 1 autoimmune hepatitis. Increased GGT values returned to pretreatment levels after 1 year in the two types. Three patients died and three other patients underwent liver transplantation. CONCLUSIONS: Non-Caucasian children had a similar disease when compared to Caucasian ones with autoimmune hepatitis. Increased levels of GGT during the first year of treatment should not be the only parameter for the indication of cholangiopathy. PMID- 16699619 TI - [Prevalence of constipation in adolescents enrolled in Sao Jose dos Campos, SP, Brazil, school's and in their parents]. AB - BACKGROUND: There is few information concerning constipation in adolescence. AIMS: To evaluate the prevalence of constipation in adolescents enrolled in schools settled down in the city of Sao Jose dos Campos, state of Sao Paulo, Brazil, and in their biological parents. PATIENTS AND METHODS: A transversal study was carried out by analyzing a convenience sample which included adolescents between 9 years and 9 months and 18 years and 7 months old and their parents in five schools situated in Sao Jose dos Campos, SP. Data was obtained from an individual and personal questionnaire filled by the father, the mother and the adolescent. Three hundred and seventy two adolescents were studied. Constipation was defined as the elimination of hard stools and the presence of at least one of the following: painful or difficult defecation, soiling or bloody stools. Scybalous feces or dry cracked stools or the defecation frequency equal or minor than two per week were considered criteria for characterization of constipation. Rome II criteria was used for evaluate the prevalence of constipation in the parents. RESULTS: The prevalence of constipation in adolescents was 22.3%, being it more frequent in female (27.4%) than in male (14.9%). Among the fathers, the prevalence of constipation (7.3%; 20/274) was lower than in the mothers. Kappa's coefficient showed a slight concordance in the concomitance of constipation in the adolescent and his/her mother (kappa = 0.12) and in the adolescent and his/her father (kappa = 0.05). CONCLUSIONS: Intestinal constipation is a prevalent disturbance in the studied group of adolescents. However, it was not found relevant concordance between constipation in the adolescents and their parents. PMID- 16699620 TI - Massive intestinal resection in rats fed up on glutamine: hepatic glycogen content valuation. AB - BACKGROUND: Glutamine has been widely used in treatment of small bowel syndrome and its metabolic effects on the small intestine are well known, however, it has been little studied its effects on hepatic metabolism under this condition. AIM: To verify through experimental model, a glutamine based supplemental diet, administered via oral to rats submitted to massive intestinal resection, evaluating weight evolution and hepatic glycogen content. MATERIAL AND METHODS: Male rats, Wistar, were allocated into three groups to undergo enterectomy. Following diets were applied: with glutamine (G group), without glutamine (NG group), and standard diet from the laboratory (R group). All animals had massive small intestine resection including ileocecal valve removal. After 20 days, all animals were sacrificed. The liver was removed to histological analysis by light microscopy. Slides were stained by periodic acid of Schiff with diastasis. RESULTS: All animals lost weight from the beginning to the end of experiment. Comparing weight loss average expressed in percentage, there was no difference statistically significant on this variance. In analyzed groups, the hepatic glycogen content did not differ statistically, in the histological method evaluated. CONCLUSION: Glutamine feeding via oral did not influence weight loss reduction of animal submitted to massive intestinal resection and did not stimulate glycogen synthesis and storage into hepatocytes. PMID- 16699621 TI - [Initial evaluation of systemic and regional pCO2 gradients as markers of mesenteric hypoperfusion]. AB - BACKGROUND: Mesenteric ischemia is a life-threatening emergency with a mortality rates still ranging between 60% and 100%. AIM: To evaluate the systemic and regional pCO2 gradients changes induced by mesenteric ischemia-reperfusion injury. In addition, we sought to determine if other systemic marker of splanchnic hypoperfusion could detect the initial changes in intestinal mucosal microcirculation after superior mesenteric artery occlusion. METHODS: Seven pentobarbital anesthetized mongrel dogs (20.6 +/- 1.1 kg) were subjected to superior mesenteric artery occlusion for 45 minutes, and followed for an additional 120 minutes. Systemic hemodynamic was evaluated through a Swan-Ganz and arterial catheters, while gastrointestinal tract perfusion by superior mesenteric vein and jejunal serosal blood flows (ultrasonic flowprobe). Intestinal oxygen delivery, extraction and consumption (DO2intest, ERO2intest and VO2intest, respectively), intramucosal pH (gas tonometry), and mesenteric arterial and mucosal arterial pCO2 gradients (D(vm-a)pCO2 and D(t-a)pCO2, respectively) were calculated. RESULTS: Superior mesenteric artery occlusion was not associated with significant changes on systemic hemodynamics parameters. A significant increase of D(vm-a)pCO2 (1.7 +/- 0.5 to 5.7 +/- 1.8 mm Hg) and D(t a)pCO2 (8.2 +/- 4.8 to 48.7 +/- 4.6 mm Hg) were detected. During the reperfusion period a significant decrease on DO2intest (67.7 +/- 9.9 to 38.8 +/- 5.3 mL/min) and a compensatory increase on ERO2intest from 5.0 +/- 1.1% to 12.4 +/- 2.7% was observed. CONCLUSION: We conclude that gas tonometry can detect the mesenteric blood flow disturbances sooner than other analyzed parameters. Additionally, we demonstrated that changes on systemic or regional pCO2 gradients are not able to detect the magnitude of intestinal mucosal blood flow reduction after mesenteric ischemia-reperfusion injury. PMID- 16699622 TI - [Nutritional evaluation of the cirrhotic patient: an objective, subjective or multicompartmental approach?]. AB - BACKGROUND: Malnutrition due to liver disease is common and its assessment is difficult. The parameters of nutritional evaluation often used in clinical practice have limited use in cirrhotic patients. Many of the clinical signs of malnutrition are the result of the underlying hepatic disease which tends to confound and alter the nutritional diagnosis. AIM: To present a brief review of the different methods of nutritional evaluation together with their limitations and uses in cirrhotic patients. CONCLUSION: The multicompartmental approach for four compartments was shown to be considerably sensitive in the detection of malnutrition. However the ample use of this method is impaired due to technical difficulties and high costs. The handgrip strength appears to be a simple, cheap and effective alternative to detected malnutrition and risk of malnutrition in these patients. Is the most sensitive method and is able to predict a significant incidence of major complications in undernourished cirrhotic patients. PMID- 16699625 TI - Distribution of virulence markers in clinical and environmental Vibrio cholerae non-O1/non-O139 strains isolated in Brazil from 1991 to 2000. AB - One hundred seventy nine Vibrio cholerae non-O1/non-O139 strains from clinical and different environmental sources isolated in Brazil from 1991 to 2000 were serogrouped and screened for the presence of four different virulence factors. The Random Amplification of Polymorphic DNA (RAPD) technique was used to evaluate the genetic relatedness among strains. Fifty-four different serogroups were identified and V. cholerae O26 was the most common (7.8%). PCR analysis for three genes (ctxA, zot, ace) located of the CTX genetic element and one gene (tcpA) located on the VPI pathogenicity island showed that 27 strains harbored one or more of these genes. Eight (4.5%) strains possessed the complete set of CTX element genes and all but one of these belonged to the O26 serogroup suggesting that V. cholerae O26 has the potential to be an epidemic strain. The RAPD profiles revealed a wide variability among strains and no genetic correlation was observed. PMID- 16699623 TI - Genetic diversity of oral Fusobacterium nucleatum isolated from patients with different clinical conditions. AB - The genetic diversity of 23 oral Fusobacterium nucleatum isolated from 15 periodontal patients, eight from seven healthy subjects, nine from nine AIDS patients and two from two Cebus apella monkeys were analyzed. EcoRI restricted the bacterial DNA and 28 ribotypes grouped from A to J groups were obtained. Isolates formed 24 ribotypes which were contained into A, B, C, D, E and F groups, and three reference strains and two clinical isolates of A. actinomycetemcomitans, and E. coli CDC formed four different ribotypes into the G, H, I and J groups. Moreover, from nine F. nucleatum from AIDS patients, six were ribotyped as group C and three as group D. By using ribotyping we distinguished F. nucleatum recovered from different sources. It is possible that isolates from AIDS patients may contain some phenotypic or genotypic factor did not observed in this study. PMID- 16699626 TI - Cryptococcus neoformans strains isolated from church towers in Rio de Janeiro City, RJ, Brazil. AB - Cryptococcosis has been a significant cause of morbidity and mortality in patients with Aids. Many reservoirs of the agent Cryptococcus neoformans have been reported, but the ecology of this yeast must be elucidated in order to establish surveillance programs and to prevent infections. The objective of this study was to evaluate the presence of C. neoformans in Rio de Janeiro City, RJ, Brazil. Ten churches were selected for sampling and detection of the yeast collecting pigeon dropping, air samples from church towers and neighboring areas during one year. The data demonstrated that C. neoformans has been present in every church selected and was present in 37.8% of 219 pigeon dropping samples. As well as, the yeast was isolated from soil, insects, eggs, pigeon nests and feathers. Fifteen air samples (4.9%) were positive. The growth on C.G.B. medium showed that all strains belonged to C. neoformans var. neoformans, with 98.8% of the strains belonging to serotype A. PMID- 16699628 TI - Mother-to-child transmission of different HIV-1 subtypes among ARV Naive infected pregnant women in Nigeria. AB - The rate of mother-to-child transmission (MTCT) of HIV as well as the implications of the circulating multiple subtypes to MTCT in Nigeria are not known. This study was therefore undertaken to determine the differential rates of MTCT of HIV-1 subtypes detected among infected pregnant women before ARV intervention therapy became available in Nigeria. Twenty of the HIV-positive women who signed the informed consent form during pregnancy brought their babies for follow-up testing at age 18-24 months. Plasma samples from both mother and baby were tested for HIV antibody at the Department of Virology, UCH, Ibadan, Nigeria. All positive samples (plasma and peripheral blood mononuclear cells PBMCs) were shipped to the Institute of Tropical Medicine, Antwerp, Belgium, where the subtype of the infecting virus was determined using the HMA technique. Overall, a mother-to-child HIV transmission rate of 45% was found in this cohort. Specifically, 36.4%, 66.7% and 100% of the women infected with HIV-1 CRF02 (IbNg), G and B, respectively, transmitted the virus to their babies. As far as it can be ascertained, this is the first report on the rate of MTCT of HIV in Nigeria. The findings reported in this paper will form a useful reference for assessment of currently available therapeutic intervention of MTCT in the country. PMID- 16699629 TI - Seroprevalence of Chagas disease in schoolchildren from two municipalities of Jequitinhonha Valley, Minas Gerais, Brazil; six years following the onset of epidemiological surveillance. AB - Six years after the beginning of the epidemiological surveillance of Chagas disease in Berilo and Jose Goncalves de Minas, Jequitinhonha Valley, MG, Brazil, a serological inquiry was performed to observe whether the transmission of this endemy was occurring in this area. A randomized sample of 1,412 children seven to 14 years old, was screened. Six asymptomatic children were found to be positive, leading to 0.4% of prevalence. Hemoculture confirmed infection in five out of the six positive cases. Additional epidemiological investigation revealed important antecedents, such as disease reports in relatives and predisposing ecological and housing conditions. Our results demonstrated similar seroprevalence (0.4%) in schoolchildren, ranging from seven to 14 years old, and that were observed six years ago (0.2%) for children 0-9 year-old. Thus, considering the constant presence of Panstrogylus megistus in the peridomicile these findings emphasize the need of continuous improved epidemiological surveillance of Chagas disease in this region. PMID- 16699630 TI - Seroprevalence for hepatitis E virus (HEV) infection among volunteer blood donors of the Regional Blood Bank of Londrina, State of Parana , Brazil. AB - A cross-sectional study was carried out among 996 volunteer blood donors enrolled from May 1999 to December 1999 to determine the seroprevalence of hepatitis E virus (HEV) infection among volunteer blood donors of the Regional Blood Bank of Londrina, State of Parana, Brazil, and to evaluate whether the rate of seroprevalence of IgG anti-HEV antibodies is associated with sociodemographic variables and with seropositivity for hepatitis A virus (HAV) infection. All participants answered the questionnaire regarding the sociodemographic characteristics. Serum samples were tested for IgG antibodies to HEV (anti-HEV) by an enzyme linked immunoassay (ELISA). All serum samples positive for anti-HEV IgG and 237 serum samples negative for anti-HEV were also assayed for IgG anti HAV antibodies by ELISA. Anti-HEV IgG was confirmed in 23/996 samples, resulting in a seroprevalence of 2.3% for HEV infection, similar to previous results obtained in developed countries. No significant association was found between the presence of anti-HEV IgG antibodies and the sociodemographic variables including gender, age, educational level, rural or urban areas, source of water, and sewer system (p > 0.05). Also, no association with seropositivity for anti-HAV IgG antibodies was observed (p > 0.05). Although this study revealed a low seroprevalence of HEV infection in the population evaluated, the results showed that this virus is circulating among the population from Londrina, South Brazil, and point out the need of further studies to define the clinical and epidemiological importance of HEV infection and to identify additional risk factors involved in the epidemiology and pathogenesis of this infection in this population. PMID- 16699632 TI - Hearing impairment in patients with tuberculosis from Northeast Brazil. AB - The aim of this paper is to describe the auditory profile of subjects who were given streptomycin treatment for tuberculosis in the years 2000 and 2001, in Recife, Northeast Brazil. The Injury Notification Database at the Municipal Department of Health was consulted and 78 individuals who had been on streptomycin during the period under study were selected. Forty-two individuals were excluded, of whom five were over 59 years of age and two were under 18 years. Nineteen turned out to be deceased, 13 could not be found, two were serving prison terms and one refused to participate. As a result, only 36 subjects participated in the study. These 36 individuals were interviewed and underwent meatoscopy and audiometry. The mean age of the group under study was 38.8 years old and males predominated (79.4%). Twenty-seven patients (75%) showed hearing impairment, the bilateral sensorineural type (63.9%) being the most frequent, mainly affecting the high frequencies over 4000 Hz. In the light of findings showing a high percentage of hearing impairment complaints among users of streptomycin, this issue seems to warrant an in-depth investigation, as does the implementation of an auditory follow-up routine for patients undergoing such chemotherapy for tuberculosis. PMID- 16699631 TI - Toxoplasma-IgM and IgG-avidity in single samples from areas with a high infection rate can determine the risk of mother-to-child transmission. AB - Anti-Toxoplasma IgG-avidity was determined in 168 serum samples from IgG- and IgM positive pregnant women at various times during pregnancy, in order to evaluate the predictive value for risk of mother-to-child transmission in a single sample, taking the limitations of conventional serology into account. The neonatal IgM was considered the serologic marker of transmission. Fluorometric tests for IgG, IgM (immunocapture) and IgG-avidity were performed. Fifty-one of the 128 pregnant women tested gave birth in the hospital and neonatal IgM was obtained. The results showed 32 (62.75%) pregnant women having high avidity, IgM indexes between 0.6 and 2.4, and no infected newborn. Nineteen (37.25%) had low or inconclusive avidity, IgM indexes between 0.6 and 11.9, and five infected newborns and one stillbirth. In two infected newborns and the stillbirth maternal IgM indexes were low and in one infected newborn the only maternal parameter that suggested fetal risk was IgG-avidity. In the present study, IgG-avidity performed in single samples from positive IgM pregnant women helped to determine the risk of transmission at any time during pregnancy, especially when the indexes of the two tests were analysed with respect to gestational age. This model may be less expensive in developing countries where there is a high prevalence of infection than the follow-up of susceptible mothers until childbirth with monthly serology, and it creates a new perspective for the diagnosis of congenital toxoplasmosis. PMID- 16699633 TI - New data on Echinococcus spp. in Southern Brazil. AB - 40 Echinococcus isolates from sheep and cattle in Southern Brazil were genetically analysed in order to obtain further data on the presence of different taxa of the Echinococcus granulosus complex. Differentiation was done using a PCR technique and sequencing of mitochondrial cytochrome c oxidase subunit 1 (CO1). Most samples (38) could be allocated to the sheep strain (G1) of E. granulosus, while two samples belonged to E. ortleppi, previously known as cattle strain (G5) of E. granulosus. Due to the shorter prepatent period in dogs of the latter taxon, this records have important implications for the design of control measures in this endemic region. PMID- 16699634 TI - Pulmonary thromboembolism in AIDS patient with chronic venous insufficiency, pulmonary tuberculosis and breast cancer: a case report and pathophysiology review. AB - Recent literature reports thrombotic episodes occurring in patients with HIV infection associated with other abnormalities including neoplasms and infections predisposing to a hypercoagulable state. We report a 47-year-old woman who developed pulmonary thromboembolism in association with HIV infection, pulmonary tuberculosis and breast cancer. She was treated with rifampin, isoniazid, pyrazinamide; heparin, phenprocoumon, zidovudine, lamivudine and efavirenz. Acid fast bacilli were visualized in a sputum smear and three months after, Mycobacterium tuberculosis was isolated from lymph node biopsy during a episode of immune reconstitution. The isolated mycobacteria showed sensitivity to all first-line drugs. HIV infection, breast cancer and pulmonary tuberculosis have several mechanisms that induce hypercoagulable state and can lead to thromboembolic complications. Pulmonary thromboembolism in this patient was a diagnostic challenge because of all the other severe diseases that she experienced at the same time. PMID- 16699635 TI - Treatment of chronic hepatitis C in non-responsive patients with pegylated interferon associated with ribavirin and thalidomide: report of six cases of total remission. AB - Hepatitis C virus (HCV) infection is an important public health issue worldwide. It is estimated that over 170 million people are infected with the virus. The present study reports six cases in which patients did not respond to combination therapy with pegylated interferon and ribavirin. However, after the addition of thalidomide to the therapy, the patients presented negative RNA PCR. The use of thalidomide combined with pegylated interferon and ribavirin for the treatment of hepatitis C is described here for the first time in the related literature. PMID- 16699636 TI - Chronic disseminated histoplasmosis with lesions restricted to the mouth: case report. AB - We report a rare case of chronic disseminated histoplasmosis with several ulcerated lesions in the oral cavity in an alcoholic patient without human immunodeficiency virus infection, with no detectable signs and symptoms of systemic disease or extraoral manifestations. Histopathological analysis revealed chronic inflammatory process with granulomas containing Histoplasma-like organisms. The isolation of Histoplasma capsulatum provided the definitive diagnosis. Treatment with itraconazole resulted in complete remission of oral lesions. As far we aware, this is the second case report of oral histoplasmosis in an HIV negative patient described in Brazil. PMID- 16699638 TI - [Factors associated with treatment failure of cutaneous leishmaniasis with meglumine antimoniate]. AB - We investigated factors associated with treatment failure in the treatment of cutaneous leishmaniasis with meglumine antimony in a reference service in Mato Grosso State. A retrospective cohort of 151 patients with cutaneous leishmaniasis was built using medical records. The incidence of therapeutic failure was 47% (IC95%=39.2%-55%). Antimoniate doses below 10 mg/kg/day (RR=1.8; IC95: 1.1-3.0), previous leishmaniasis treatment (RR=1.7; IC95: 1.3-2.4), 3 or more skin lesions (RR=1.9; IC95: 1.4-2.5), incomplete treatment (RR=1.9; IC95: 1.3-2.6) and body weight above 68 kg (RR=1.7; IC95: 1.1-2.5) were associated with therapeutic failure. After adjustment, therapeutic failure was associated with having 3 or more cutaneous lesions (OR=4.6; IC95%=1.2-17.4), reports of previous leishmaniasis treatment (OR=4.5; IC95%=1.1-17.5), body weight above 68 kg (OR=4.3; IC95=1.5-11.9) and incomplete treatment schedule (OR=12.5; IC95%=2.1 75.4), although body weight is possibly associated with treatment failure due to the limitation of the maximum daily dose. These results help to identify patients at risk of treatment failure of cutaneous leishmaniasis with antimony therapy. PMID- 16699639 TI - Neurological disease in HIV-infected patients in the era of highly active antiretroviral treatment: a Brazilian experience. AB - To study characteristics of neurological disorders in HIV/AIDS patients and their relationship to highly active antiretroviral treatment, a cross-sectional study was conducted in an infectious disease public hospital in Belo Horizonte, Brazil, between February 1999 and March 2000. Of the 417 patients enrolled, neurological disease was observed in 194 (46.5%) and a new AIDS-defining neurological event developed in 23.7% of individuals. Toxoplasmosis (42.3%), cryptococcosis meningitis (12.9%) and tuberculosis (10.8%) were the most common causes of neurological complications. The majority (79.3%) of patients were on highly active antiretroviral treatment and these individuals using HAART showed higher CD4 cell counts (p = 0.014) and presented stable neurological disease (p = 0.0001), although no difference was found with respect to the profile of neurological complications. The neurological diseases continue to be a frequent complication of HIV/AIDS and infections are still its main causes in Brazil, even in the highly active antiretroviral treatment era. PMID- 16699640 TI - [Radiographic study of the oesophagus of chagasic patients in 25 years of the Mambai Project]. AB - This work describes radiography studies of the esophagus in chagasic patients of the Mambai project over a 25 year period. Each person realized two radiographs in four different periods (1975-1976, 1980-1982, 1988-1991 and 1998-2000). The first radiograph was realized in the right anterior oblique position after swallowing 75 ml of barium sulphate solution and another 60 seconds later. In all periods the same methodology was used. The radiographs were read by the same researcher, and the megaesophagus was classified in four groups according to Rezende and colleagues. The prevalence of megaesophagus was 5.2%; 5.0%; 18.6% and 13.9% in 1975-1976, 1980-1982, 1988-1991 and 1998-2000 respectively. The incidence of megaesophagus from 1975 to 2000 was 11.5% (51/445). During 25 year, 394 (84.2%) patients presented radiographs normal of esophagus, 11 (2.3%) diagnosed as megaesophagus in 1975-1976 did not progress, 61 (13%) that were normal, doubtful or already presented megaesophagus in 1975-1976, progressed and 2 (0.4%) presented regression of group I megaesophagus, diagnosed in previous studies. PMID- 16699641 TI - [Manometric evaluation of upper esophageal sphincter in patients with indeterminate form of Chagas' disease]. AB - The objective was to study the disorders of upper esophageal sphincter in 37 patients with indeterminate clinical form of Chagas' disease. Eighty (48.6%) patients with synchronic waves were found. The average maximum pressure value of the upper esophageal sphincter was significantly higher in the synchronic group. Subjects with indeterminate clinical form of Chagas' disease may have functional disorders demonstrated by an increase in the pressure of the upper esophageal sphincter. PMID- 16699642 TI - Digoxigenin-labeled probe for rabies virus nucleoprotein gene detection. AB - A digoxigenin-labeled probe was produced from the Pasteur virus strain for the detection of the rabies virus N gene. The probe hybridization was performed from amplified N gene obtained by reverse transcription polymerase chain reaction and the results by RT-PCR and hybridization showed 100% agreement. The hybridization, when carried out in products amplified by RT-PCR, increases the sensitivity of this technique even more and confers specificity to the diagnosis. The technique described in this work will be useful in rabies diagnosis laboratories, once the cost is compatible with traditional rabies diagnostic techniques. PMID- 16699643 TI - [Epidemiologic profile of puerperals and prevalence of antibodies for HIV and hepatitis C virus infection in Cuiaba, Mato Grosso]. AB - A seroepidemiologic survey was performed aimed at estimating the prevalence of human immunodeficiency virus, the prevalence of hepatitis C virus antibodies (anti-HCV), the risk factors associated with human immunodeficiency virus transmission and the epidemiological profile of the women in puerperal period in three public Hospitals of Cuiaba City between December 2001 and March 2002. The sample was composed of 1,607 women who were interviewed in order to obtain information about socio-demographic aspects and risk behavior. Blood samples were drawn to detect HIV and HCV antibodies using the ELISA test. The prevalence of HIV infection in this population was 0.5% (IC95%= 0.2; 1.0). Most women had only basic level schooling (58.4%), and they presently held a relationship with a fix partner (73%). There were no statistically significant associations between HIV antibody and socioeconomic level, other procedures that involved the risk of HIV parenteral transmission, between anti-HIV positivity and the presence of sexual behavior of risk (relationships with multiple partners or bisexuality) by the women and their partners. Currently, it is presumed that heterosexual transmission is the main cause of infection among women in reproductive age in the study area. The anti-HCV prevalence was 0.4% (CI95%= 0.1; 0.8). Anti-HCV was more common among older women. PMID- 16699644 TI - [Cloning and transmembrane glycoprotein expression of the retrovirus HTLV-1 in mammals' cells]. AB - The retrovirus HTLV-1 is the etiological agent of the adult T-cell leukemia and HTLV-1 associated myelopathy/tropical spastic paraparesis. The proviral genome has 9,032 base pairs, showing regulatory and structural genes. The env gene encodes for the transmembrane glycoprotein gp 21. The development of methodologies for heterologous protein expression, as well as the acquisition of a cellular line that constituently expresses the recombinant, were the main goals of this work. The DNA fragment that encodes for gp 21 was amplified by nested-PCR and cloned into a pCR2.1-TOPO vector. After which, a sub-cloning was realized using the expressing vector pcDNA3.1+. The transfection of mammalian cells HEK 293 was performed transitorily and permanently. Production of the recombinant gp 21 was confirmed by flux cytometry experiments and the cell line producing protein will be used in immunogenicity assays. PMID- 16699645 TI - Differentiation of Candida species obtained from nosocomial candidemia using RAPD PCR technique. AB - Thirteen strains of the genus Candida were isolated from catheter, urine and surgical wounds from individual patients of the Santa Casa de Misericordia, Belo Horizonte, MG, Brazil. Ten strains were characterized as Candida albicans, two as Candida glabrata, and one as Candida parapsilosis. Isolates were evaluated for molecular relatedness by random amplified polymorphic DNA technique using 15 primers. The analysis of the genomic DNA obtained revealed a low intraspecific polymorphism and did not allow for the differentiation between strains of the same species obtained from distinct clinical sources (catheter, urine and surgical wounds). The RAPD profiles generated were able to differentiate among the species of Candida albicans, Candida parapsilosis and Candida glabrata strains isolated in this study. PMID- 16699646 TI - [Evaluation of the sorologic pre-selection for the hepatitis B virus marker (total anti-HBc) in candidates to blood donation in the State of Acre, 2002]. AB - The aim of this research was to evaluate previous serum screening for hepatitis B (total anti-HBc) among blood donation candidates, verifying the link between the variables: gender, age, degree of education and native-born. A transverse study with retrospective data carried out on a target population, blood donation candidates from districts of the interior of Acre State, who came to the Acre Hematology and Hemoterapy Center during the period January to December, 2002. The research indicated reactivity in 54.8% of the total anti-HBc among the 673 candidates included. Greater reactivity of total anti-HBc was observed among males, older candidates and those with lower education levels (p<0.005). Previous serum screening for hepatitis B among blood donation candidates is a viable alternative, since it reduces cost and increases transfusional safety. Attracting female donors, youths and those who have higher education levels, would signify potential blood donors for HEMOACRE. PMID- 16699647 TI - [Blood meals identification of Lutzomyia (Lutzomyia) longipalpis (Lutz & Neiva, 1912) e Lutzomyia (Lutzomyia) almerioi (Galati & Nunes, 1999) by enzyme-linked immunosorbent assay biotin-avidin]. AB - Lutzomyia longipalpis and Lutzomyia almerioi, phlebotomine species from the fauna of Serra da Bodoquena, in the State of Mato Grosso do Sul, Brazil, have been studied, particularly due to the fact of their abundance and occurrence, the Guaicurus settlement, focus of human tegumentary and canine visceral leishmaniasis. In researches that are being carried out in this settlement for identifying the vectors of these parasitosis, 83 engorged females belonging to the species Lutzomyia longipalpis and Lutzomyia almerioi were captured with automatic light traps from 2002 up to 2004 in the peridomiciliary environment of the Guaicurus settlement (hennery). The aim of this study was the investigation on bird feeding habit of females of both the phlebotomine species by the enzyme linked immunosorbent assay technique, comparing the reactivity during the period from 2002 up to 2004. Of the 57 samples of Lutzomyia longipalpis and 26 of Lutzomyia almerioi that have been tested, 72% from Lutzomyia longipalpis and 96% from Lutzomyia almerioi were reactive, which justifies the feeding habit study in the region as a prevention measure and the institution of an epidemiological survey. PMID- 16699648 TI - [Epidemiological trends of malaria in the State of Mato Grosso, from 1980 to 2003]. AB - The temporal and spatial evolution of malaria in Mato Grosso was determined in periods from 1980-1985, 1986-1991, 1992-1997 and 1998-2003 and distributed by homogeneous microregion. The annual parasitic index of the state rose until 1992 and then diminished to 1.9 cases/1,000 inhabitants in 2003, the ratio of mortality and the lethality rate were greater in the 1980s. Of the 22 microregions, 13 presented an API inferior to 10 cases/1,000 inhabitants in all periods. Cases were concentrated in the microregions of Colider, Alta Floresta, Aripuana and Alto Guapore. In 2003, only the microregion of Aripuana continued to present an API superior to 50 cases/1,000 inhabitants. The microregions of Colider, in 1983, 1985 to 1988 and 1990 and Alta Floresta, in 1991 presented deaths over 50/100,000 inhabitants, mainly in males aged 20-49 years. The distribution of the disease in microregions showed that malaria is predominantly found in concentrated sites. PMID- 16699649 TI - Experimental ophitoxemia produced by the opisthoglyphous lora snake (Philodryas olfersii) venom. AB - Several colubrid snakes produce venomous oral secretions. In this work, the venom collected from Venezuelan opisthoglyphous (rear-fanged) Philodryas olfersii snake was studied. Different proteins were present in its venom and they were characterized by 20% SDS-PAGE protein electrophoresis. The secretion exhibited proteolytic (gelatinase) activity, which was partially purified on a chromatography ionic exchange mono Q2 column. Additionally, the haemorrhagic activity of Philodryas olfersii venom on chicken embryos, mouse skin and peritoneum was demonstrated. Neurotoxic symptoms were demonstrated in mice inoculated with Philodryas olfersii venom. In conclusion, Philodryas olfersii venom showed proteolytic, haemorrhagic, and neurotoxic activities, thus increasing the interest in the high toxic action of Philodryas venom. PMID- 16699650 TI - [Rupestrian triatomines infected by Trypanosomatidae, collected in Quarai, Rio Grande do Sul, 2003]. AB - Rupestrian triatomines were captured in six Quarai city localities, RS, to verify the level of Trypanosomatidae infection, as well as the animal reservoir. The capture occurred in a wild environment and 453 samples were collected, which were identified and separated by nymphal instar. 421 (92.9%) samples of Triatoma rubrovaria, 26 (5.7%) of Triatoma circummaculata and 6 (1.3%) of Panstrongylus tupynambai were collected. Only 13 samples (4.2%) of Triatoma rubrovaria presented Trypanosomatidae infection. After mice and LIT culture inoculation, five strains of Trypanosoma cruzi were isolated. Of these triatomines, 4 (30.8%) displayed no reagent precipitin for the antiserum tested, 4 (30.8%) were positive for rodent antiserum, 4 (30.8%) were positive for goat antiserum and 1 (7.7%) were positive for human and pig antiserum. PMID- 16699651 TI - [Viral hemorrhagic fevers in Brazil]. AB - To call attention to viral hemorrhagic fevers, diseases that are mostly underdivulged and, probably, underreported, we present here case reports of the 4 diseases of this kind that occur in Brazil: yellow fever, dengue haemorrhagic fever/dengue shock syndrome, arenavirus haemorrhagic fever and hantavirus cardiopulmonary syndrome. Relevant clinical, epidemiological and laboratorial diagnostic aspects of these viral haemorrhagic fevers are also shown here. These diseases have a high case fatality rate, induce capillary leaking and blood coagulation disturbances that are evidenced by hemoconcentration and thrombocytopenia. An early clinical diagnosis and treatment is fundamental for patient survival. PMID- 16699652 TI - [Chronic chagasic cardiopathy in the Rio Negro, Amazon State. Report of three new autochthonous cases confirmed by serology, clinical examination, chest X-rays, electro and echocardiography]. AB - We report three new autochthonous cases of chronic chagasic cardiomyopathy from Rio Negro, Amazon State, confirmed by serology (indirect immunofluorescence, ELISA and Western-blot for Trypanosoma cruzi infection), clinical examination, chest X-rays, electro- and echocardiography. The three patients were born and lived all their lives in the Rio Negro region working as piacaba gatherers, where they were bitten several times by sylvatic triatomine bugs. The clinical feature was congestive heart failure and intraventricular conduction impairment in the three cases (right bundle branch block with left anterior hemiblock in two cases, left bundle branch block of 3rd degree in one), polymorphic ventricular extrasystoles in two cases and primary T wave inversion in another one. The echocardiographic evaluation showed a significant increasing of the left ventricular diameters with ejection fraction lesser than 36%, and myocardial segmental impairment pattern, including apical aneurysm and postero-inferior akinesia in the three patients. These are the first autochthonous cases of chronic Chagasic cardiomyopathy from the Brazilian Amazon state with echocardiographic pattern suggestive of Chagas' disease. PMID- 16699653 TI - [Aeromonas associated with an acute diarrhea outbreak in Sao Bento do Una, Pernambuco]. AB - An acute diarrhea outbreak, with 2170 cases, was described during January to July, 2004, in Sao Bento do Una, Pernambuco. 582 stools were examined and an enteric pathogen was recovered in 25% (145 patients). Aeromonas species were the most frequent (114-19.5%) and the main isolates were Aeromonas caviae (57-9.8%), Aeromonas veronii biovar sobria (23-3.9%), Aeromonas veronii biovar veronii (15 2.6%) and other species (19-3.2%). The other isolated enteropathogens were Vibrio cholerae O1-Ogawa toxigenic (18-3.1%), Salmonella spp (8-1.4%), Shigella spp (3 0.5%) and Vibrio cholerae non-O1/non-O139 (2-0.3%). PMID- 16699654 TI - [Radiographic patterns of pulmonary tuberculosis among the Surui indians of Rondonia, Amazonia]. AB - The radiographic patterns of 23/33 (69.7%) Surui indians, of the Brazilian Amazon, treated in 2003-2004 were analyzed. The analysis showed that 44.8% presented unspecific infiltrates, 10.3% presented cavitations and 39.1% presented multiple parenchymal involvements. Although 36% of the patients presented normal radiographs, they underwent anti-tuberculous treatment without having been submitted for other possible means of diagnosis. PMID- 16699655 TI - [Analysis of the vegetable productive chain in Ribeirao Preto, SP]. AB - With the aim of assessing the cumulative risk of lettuce contamination, 45 production chains were investigated. The presence of thermotolerant coliforms, Salmonella and/or parasites was detected in 69% of these, in all steps of the production chain. Quality control in all steps of the lettuce production process should be intensified. PMID- 16699656 TI - [Integral insecticide spraying followed by immediate triatomine research, as an additional strategy for the epidemiological surveillance of Chagas' disease, in areas of residual persisting foci]. AB - The locality of Altolandia (Tapirai Municipality, Minas Gerais, Brazil), frequently presents Panstrongylus megistus infestation in the domiciliary environment. In order to improve epidemiological surveillance in this area, a strategy of domiciliary pyrethroid spraying with high flushing-out effect followed by immediate triatomine research, was implemented in all the existing dwellings. The main objectives were the detection and elimination of cryptic foci, thus reducing the opportunities for new infestations. PMID- 16699657 TI - [Importance of culture of urine in the diagnosis of AIDS associated cryptococcosis]. AB - In this retrospective study we analyzed 70 HIV patients with cryptococcosis over a 16-years period. The specimens with the best positivity were CSF (97.8%) followed by the culture of urine sediment (86.7%) and blood culture (58.8%). We conclude that the urine could be a useful tool for the diagnosis of cryptococcosis. PMID- 16699658 TI - Schistosomiasis mansoni of the prostate. PMID- 16699660 TI - [Full Threshold vs. SITA in glaucomatous patients undergoing automated perimetry for the first time]. AB - PURPOSE: To compare the Full Threshold (FT) and SITA Standard (SS) strategies in glaucomatous patients undergoing automated perimetry for the first time. METHODS: Thirty-one glaucomatous patients who had never undergone perimetry underwent automated perimetry (Humphrey, program 30-2) with both FT and SS on the same day, with an interval of at least 15 minutes. The order of the examination was randomized, and only one eye per patient was analyzed. Three analyses were performed: a) all the examinations, regardless of the order of application; b) only the first examinations; c) only the second examinations. In order to calculate the sensitivity of both strategies, the following criteria were used to define abnormality: glaucoma hemifield test (GHT) outside normal limits, pattern standard deviation (PSD) <5%, or a cluster of 3 adjacent points with p<5% at the pattern deviation probability plot. RESULTS: When the results of all examinations were analyzed regardless of the order in which they were performed, the number of depressed points with p<0.5% in the pattern deviation probability map was significantly greater with SS (p=0.037), and the sensitivities were 87.1% for SS and 77.4% for FT (p=0.506). When only the first examinations were compared, there were no statistically significant differences regarding the number of depressed points, but the sensitivity of SS (100%) was significantly greater than that obtained with FT (70.6%) (p=0.048). When only the second examinations were compared, there were no statistically significant differences regarding the number of depressed points, and the sensitivities of SS (76.5%) and FT (85.7%) (p=0.664). CONCLUSION: SS may have a higher sensitivity than FT in glaucomatous patients undergoing automated perimetry for the first time. However, this difference tends to disappear in subsequent examinations. PMID- 16699661 TI - [MALKS: specifications and development]. AB - PURPOSE: To describe the characteristics of a new artificial anterior chamber (MALKS, Loktal, Sao Paulo, Brazil). METHODS: Characteristics analysis and description of a new artificial anterior chamber MALKS (Micro automated lamellar keratoplasty system). RESULTS: MALKS is composed of eight parts: a) artificial chamber; b) cornea's fix ring; c) nut to join the cornea's fix ring; d) rail and adjuster of lamellar diameter; e) flatteners, to pre-determine lamellar diameter; f) infusion system, that allows the digital objective peroperative control of the intracameral pressure; g) automated microkeratome, and h) marker. CONCLUSION: MALKS uses the same automated microkeratome developed for LASIK, can allow corneal lamella obtention with predetermined thickness and diameter, as well as the digital objective peroperative control of the intracameral pressure. This new artificial anterior chamber can be an important tool for superficial and endothelial keratoplasty. PMID- 16699662 TI - [Comparative analysis of macular hole surgery followed by internal limiting membrane removal with and without indocyanine green staining]. AB - PURPOSE: To describe comparatively the outcomes of macular hole surgery followed by internal limiting membrane removal with and without indocyanine green staining. METHODS: We evaluated 142 consecutive macular hole surgeries with internal limiting membrane removal performed in the period from January 2001 to March 2004. These were divided into two groups, based on the use or not of indocyanine green staining. RESULTS: Both studied groups were similar concerning the patients' profile and preoperative stage of the macular holes. Mean preoperative visual acuity was 0.12+/-0.15 in the group with indocyanine green staining and 0.18+/-0.18 in the group without internal limiting membrane staining (p=0.02). Mean postoperative visual acuity was 0.27+/-0.27 in the group with internal limiting membrane staining and 0.43+/-0.25 in the group without internal limiting membrane staining (p=0.0002). We observed improvement of visual acuity in 63% of cases in the group in which internal limiting membrane staining was performed and in 80.3% of cases in the group without internal limiting membrane staining. Macular hole closure rates were 76.5% in the group treated with internal limiting membrane staining and 95.1% in the group treated without internal limiting membrane staining. CONCLUSIONS: Macular hole surgeries followed by internal limiting membrane removal without use of staining have better visual and anatomic results when compared to macular hole surgeries followed by internal limiting membrane removal guided by indocyanine green staining. We recommend caution in using indocyanine green staining in macular hole surgeries due to its possible toxic effect. PMID- 16699663 TI - Matrix metalloproteinase-9 expression in pterygium. AB - PURPOSE: To assess matrix metalloproteinase (MMP)-9 expression in pterygium lesion. METHODS: A prospective randomized clinical trial was done to evaluate the expression of matrix metalloproteinase in normal and in primary or recurrent pterygia in Tenon's capsule by immunohistochemical analysis and a computerized image analysis system. The data were submitted to statistical analysis. RESULTS: Matrix metalloproteinase expression showed no difference in normal Tenon's capsule and in primary or recurrent pterygia. CONCLUSION: The similar expression of the matrix metalloproteinase in normal Tenon's capsule and in primary or recurrent pterygia allowed us to conclude that matrix metalloproteinase is not implicated in the genesis or the recurrence of pterygium lesion. PMID- 16699664 TI - [Evaluation of central serous chorioretinopathy with optical coherence tomography]. AB - PURPOSE: To evaluate optical coherence tomography as an objective diagnostic and follow-up method for central serous chorioretinopathy. METHODS: Observational descriptive study of 16 (sixteen) eyes of 16 patients with unilateral acute central serous chorioretinopathy. These patients had their visual acuity measured and complete ophthalmological examination was performed, including macular slit lamp examination, angiofluoresceinography and optical coherence tomography. No treatment was given to any patient. RESULTS: Of the 16 central serous chorioretinopathy eyes, 12 patients completed follow-up until total resolution of their disease. Age varied between 27 and 50 years, with mean age of 38.9 years. Most patients were male. On optical coherence tomography, neurosensorial retinal detachment was observed in all cases and 33.3% had retinal pigmented epithelium detachment. There was a significant difference regarding all studied variables. Visual acuity was better, macular thickness and volume diminished when the disease resolved. CONCLUSION: Optical coherence tomography showed good efficacy to detect and measure macular changes in eyes with central serous chorioretinopathy, being useful in clinical evaluation during the disease's acute period and resolution. PMID- 16699665 TI - [Phacoemulsification of first and second eye: indication criteria and results]. AB - PURPOSE: To compare indication criteria, intra- and postoperative complications, visual outcomes and time interval between the first and second eye surgery (phacoemulsification). METHODS: Prospective study was done in 96 eyes of patients who underwent bilateral consecutive phacoemulsification surgery, performed by a third-year ophthalmology resident. An established protocol was filled out before, during and after the surgery. The same surgical technique was used. Chi-squared, Mann-Whitney, Kruskal-Wallis, Dunn multiple comparison and Student's t tests were used for statistical analysis. Bicaudal tests were used in all situations. p<0.05 rejected the null hypothesis. RESULTS: The indication for the second eye surgery was done earlier and with best correct visual acuity when compared with the first eye (p=0.016). The mean phacoemulsification ultrasound time (US time) of the first eye was significantly higher (p=0.026). There were no statistically significant differences between the two groups in relation to type of cataract, previous intraocular diseases, final visual acuity and mean spherical equivalent. The incidence of intra- and postoperative complications was smaller in the second eye, but was not statistically significant (p=0.07). Time interval between the first and second eye surgery was significantly lower along the trimesters. CONCLUSION: The patients underwent the second eye phacoemulsification surgery earlier (with better visual acuity and shorter time interval), had a shorter phacoemulsification time and tendency to be followed by less intra- and postoperative complications. PMID- 16699666 TI - [Ocular manifestation in patients with tuberculosis]. AB - PURPOSE: To analyze clinical findings of ocular tuberculosis and its epidemiological features. METHODS: Were reviewed clinical files of patients who attended the Ophthalmology-Uveitis and AIDS sector of UNIFESP-EPM, Sao Paulo, Brazil between January 1999 and July 2002 and had a diagnosis of ocular tuberculosis. Patients who had Mantoux test higher than 10 mm, epidemiological history of tuberculosis and ocular findings that improved after specific treatment were included. RESULTS: Seventeen medical files were analyzed, of those, 12 (70.6%) were females. The mean age was 54 (24-84) years. The average time between symptoms and diagnosis was 100 days. Ocular findings at the first interview were multiple but most were at the anterior segment of the eye and 41% of patients who had clinical findings that suggested tuberculosis other than ocular. Situations as glaucoma, retinal detachment and vitreous hemorrhage were responsible for reduction of visual acuity. CONCLUSIONS: Ocular tuberculosis showed a low prevalence but important visual loosing morbidity. The multiplicity of clinical findings is responsible for the delay of diagnosis. PMID- 16699668 TI - [Bipedicle myocutaneous flap for cicatricial ectropion correction]. AB - PURPOSE: To evaluate the correction of lower eyelid cicatricial ectropion due to shrinking of anterior lamela with the surgical technique of bipedicle myocutaneous flap from the upper eyelid. METHODS: Prospective study with eight eyelids of six patients. There were two patients with unilateral ectropion who had received external local radiotherapy. RESULTS: There were three male and three female patients, with mean age of 72 years. One patient presented an incomplete blinking for one month that spontaneously resolved. Three eyelids of two patients maintained a residual ectropion; one of them had received local external radiotherapy. Follow-up ranged from 19 to 23 months (mean of 20 months) and all patients improved their ocular condition. CONCLUSION: The bipedicle myocutaneous flap is effective for correction of cicatricial ectropion due to shrinking of anterior lamella. PMID- 16699667 TI - [Strabismic amblyopia: a retrospective study on patients from a university hospital]. AB - PURPOSE: To evaluate the influence of compliance with treatment, severity of amblyopia and age at the beginning of treatment in patients with strabismic amblyopia submitted to patching. METHODS: The data were selected from medical records of 569 patients seen at the Department of Ophthalmology, University of Sao Paulo during the period of 1983 to 2000. EXCLUSION CRITERIA: loss of follow up, age over 12 years, presence of nystagmus and other ocular diseases. The patients were divided into four groups according to age and classified based on the type of strabismus, compliance with treatment and severity of amblyopia. Statistical analysis was performed using Fisher's test. RESULTS: 198 (34.8%) patients were enrolled in the study. There was no difference regarding compliance among the age groups. The best success rate was obtained when there was good compliance with the treatment, independent of the degree of amblyopia. But worse compliance was observed among the patients with more severe amblyopia, who were the majority of patients of the study, and who showed the worst success rate. The success rate was not related to the patients' age. CONCLUSION: The study showed that compliance with patching plays a fundamental role in therapeutic efficacy. Thus, age at the beginning of treatment alone did not influence the success rate since it was possible to obtain good compliance independent of age. In addition, worse results were associated with severe amblyopia and less compliance. PMID- 16699669 TI - [Prostaglandin analogues reduce the ibopamine provocative test specificity in glaucoma]. AB - PURPOSE: To evaluate the ibopamine provocative test for the diagnosis of glaucoma in glaucoma patients using antiglaucomatous drugs. METHODS: Two 2% ibopamine eyedrops were instilled 5 minutes apart in one eye selected at random in both glaucoma and normal subjects. The intraocular pressure (IOP) was assessed prior to the drops and 30, 60 and 180 minutes after instillation. The test was considered positive when there was an intraocular pressure increase of greater than 4 mmHg at any one of the time-points. The amount of intraocular pressure change was compared to the types of medical treatment. RESULTS: Fifty-eight eyes were included (38 glaucoma patients and 20 normal individuals). The intraocular pressure rise was significantly higher in glaucoma patients (p<0.001 at all times). The sensitivity and specificity of the ibopamine test were 68% (87% if we exclude eyes using prostaglandin analogues) and 95%, respectively. Glaucoma patients using prostaglandin analogues did not present a significant intraocular pressure elevation. CONCLUSION: The ibopamine provocative test may be an auxiliary test in glaucoma diagnosis. Despite the small sample size, concurrent use of prostaglandin analogues apparently reduces the test's sensitivity. PMID- 16699670 TI - [Profile of medical students from the Universidade Estadual Paulista-UNESP- Botucatu, who wear contact lenses]. AB - PURPOSE: To analyze the ametropy, way of acquisition, the care and complications related to contact lens wear among medical students from Faculdade de Medicina de Botucatu da Universidade Estadual Paulista-UNESP--Botucatu (SP). METHODS: Questionnaire applied to students from the 1st to the 6th year of the Medical school of the Faculdade de Medicina de Botucatu da Universidade Estadual Paulista UNESP--Botucatu (SP), forming an analytical cross-sectional survey. RESULTS: 27.4% of the students use contact lenses, and among them 94.8% use soft contact lenses and 98% have negative degree. 46.9% discard their contact lens monthly. 63.9% have an average daily use of 14 hours, 14.1% acquire the contact lenses from optician stores or at pharmacies and 46.5% of the students clean their contact lenses when they put them on and take them off. Regarding complications, 51.0% reported ocular redness; 44.5%, blurred vision; and 42.6%, foreign body sensation, whereas only 4 students reported "keratitis". CONCLUSION: We concluded that the inadequate adaptation that occurs even in users knowledgeable about this issue such as medical students, has increased the risk of complications. This fact indicates that more attention must be paid to contact lens care in order to prevent permanent visual deficit. PMID- 16699671 TI - [Fundus changes evaluation in degenerative myopia]. AB - PURPOSE: To evaluate the presence of fundus changes in degenerative myopia. METHODS: Forty patients with refractive error of at least -6.00 diopters were selected for ophthalmologic examination followed by posterior pole photograph and echobiometry. RESULTS: We studied 57 eyes of 37 patients with refractive error ranging from -6.25 to -28.50 diopters, with mean -14.05, and axial length ranging from 26.06 to 32.86 mm, with mean 28.01. We found a temporal crescent in 36.5% and a peripapillary one in 20% of the eyes. Choroidal vessels were seen in 35% of the eyes. Posterior pole changes were as follow: posterior staphyloma in 10.5%, Fuchs' spots in 3.5% and lacquer cracks in 1.5%. Peripheral retinal examination revealed paving stone chorioretinal atrophy in 17.5%, white without pressure in 10.5%, lattice degeneration in 5%, retinal tears in 3.5% and retinoschisis in 1.5% of the examined eyes. CONCLUSIONS: Fundus changes that lead to decreased vision are common in patients with degenerative myopia. Peripheral retinal examination is very important in these patients due to the increased risk of retinal detachment. PMID- 16699672 TI - [Utilization of optical devices and equipments by students with visual impairment]. AB - PURPOSE: To verify perceptions and conduct of students with visual impairment regarding devices and equipment utilized in schooling process. METHODS: A transversal descriptive study on a population of 12-year-old or older students in schooling process, affected by congenital or acquired visual impairment, inserted in the government teaching system of Campinas during the year 2000. An interview quiz, created based on an exploratory study was applied. RESULTS: A group of 26 students, 46% of them with low vision and 53.8% affected by blindness was obtained. Most of the students were from fundamental teaching courses (65.4%), studying in schools with classrooms provided with devices (73.1%). Among the resources used in reading and writing activities, 94.1% of the students reported they used the Braille system and 81.8% reported that the reading subject was dictated by a colleague. Most of the students with low vision wore glasses (91.7%), and 33.3% utilized a magnifying glass as optical devices. Among the non optical devices, the most common were the environmental ones, getting closer to the blackboard (75.0%) and to the window (66.7%) for better lighting. CONCLUSIONS: It became evident that students with low vision eye-sight made use of devices meant for bearers of blindness, such as applying the Braille system. A reduced number of low vision students making use of optical and non-optical devices applicable to their problems were observed, indicating a probable unawareness of their visual potential and the appropriate devices to improve efficiency. PMID- 16699673 TI - [Ophthalmological service quality offered to outpatients of the Public Healthcare System]. AB - PURPOSE: To identify the perception of the ophthalmic service quality provided for outpatients of the public healthcare system as well as to detect which actions should be considered necessary and priority in order to improve its quality. METHODS: A quantitative descriptive study was carried out on 100 outpatients of the public healthcare system which were submitted to ophthalmic tests at Fundacao Hilton Rocha--Belo Horizonte-MG, from July 1st-July 30th 2004. Individual interviews were carried out by giving the interviewees two structured questionnaires adapted from the modified SERVQUAL. This scale is in agreement with the reality of the studied institute. RESULTS: The adapted SERVQUAL scale was submitted to statistical validation and it showed a suitable internal consistency index. In general terms, a slight general dissatisfaction was detected regarding ophthalmological service quality. The interviewees cared more about safety and reliability. A higher degree of dissatisfaction was detected mainly concerning fulfillment of procedures at scheduled appointments related to the execution of services within due time-limits. CONCLUSIONS: The institute is supposed to plan as well as carry out actions which lead to a general improvement in the patient's satisfaction regarding service quality and mainly reliability. Service quality monitoring through periodic use of the SERVQUAL scale will not only make it possible to plan highly precise and effective intervention strategies in these and in other healthcare services but it will also allow monitoring the responses to these actions. All these actions will contribute to the improvement of the service in the system as a whole. PMID- 16699674 TI - [Development of a low vision aid device]. AB - PURPOSE: To develop the design of a near low vision aid that has esthetic and functional qualities intending to give its user a better life quality. METHODS: To verify low vision people's main needs concerning the current devices for near distance; to identify possible design interventions to make a project that fulfills their main needs. RESULTS: A prototype having a reading stand tied to the end of a magnifier that moves horizontally and vertically permitting the horizontal reading of a text line was developed. CONCLUSIONS: The developed device fulfills low vision people's visual and handling needs for near distance, contributing to the easiness to read and write. PMID- 16699675 TI - [Evaluation of dry eye in before and after blepharoplasty]. AB - PURPOSES: To evaluate the importance of objective examinations together with ocular history and orbital and periorbital anatomy, to identifying patients at risk of developing a post-blepharoplasty dry eye complication. METHODS: A prospective study was performed on 29 patients with dermatochalasis that were studied before and three months after blepharoplasty. Changes in ocular symptoms, physical examination and in objective tests, like Schirmer's test, tear film break-up time and rose bengal coloration were evaluated. RESULTS: No statistical significant alteration in results of objective tests, except Schirmer II test, between pre and postoperative period were found. In relation to eyelid anatomy, alterations in palpebral fissure and in excursion of the upper eyelid, that were statistically significant were found after surgery. CONCLUSIONS: It may be stated that palpebral fissure alterations occur frequently in patients submitted to esthetic blepharoplasty. It is also possible to conclude, that in this study, the objective tests for dry eye evaluation, did not demonstrate significant postoperative alterations, except for Schirmer II test. PMID- 16699676 TI - [Ophthalmological alterations in outpatients with systemic lupus erythematosus]. AB - PURPOSE: To evaluate the ophthalmological alterations in patients with systemic lupus erythematosus (SLE). METHODS: A descriptive cross-sectional study was conducted to analyze 70 patients with diagnosis of SLE based on the American College of Rheumatology criteria. Data regarding sex, age, ethnicity, duration of disease, disease activity, use of corticosteroids and antimalarial drugs, and presence of ophthalmological symptoms were recorded and a full ophthalmic examination was carried out. RESULTS: Sixty-two patients (88.6%) were females, 43 patients (61.4%) were white. Mean age was 43.9 years, 57 patients (81.4%) presented no sign of active disease and 46 patients (65.7%) had no ophthalmological complaints. Dry eye syndrome was diagnosed in 22 patients (31.4%), bilateral subcapsular cataract in 15 patients (21.1%) and glaucoma in 2 patients (2.9%). Fundoscopic evaluation disclosed 8 patients (11.4%) with drusen, 7 (10.0%) with RPE atrophy, 2 patients (2.9%) presented pathological disc excavation, 2 (2.9%) vascular tortuosity and arteriovenous crossing changes, 1 patient (1.4%) had macular chorioretinitis scar and 1 patient (1.4%) had branch retinal vein occlusion. CONCLUSION: SLE is a disease that can present with ophthalmic involvement, usually benign in outpatients. Dry eye and cataract are the commonest ocular findings. The former seems to be more related to SLE itself while the latter probably relates to the chronic use of corticosteroids for the disease treatment. Antimalarial drugs do not frequently cause ophthalmic lesions, although further studies must be conducted in order to assess their impact on patients' long-term visual quality. In the patients with the disease under control, ocular damage is more related to systemic treatment what enhances the need for ophthalmological examination as a routine even in these patients. PMID- 16699677 TI - Quantitative comparison of different-shaped wavefront sensors and preliminary results for defocus aberrations on a mechanical eye. AB - PURPOSE: There is a general acceptance among the scientific community of Cartesian symmetry wavefront sensors (such as the Hartmann-Shack (HS) sensor) as a standard in the field of optics and vision science. In this study it is shown that sensors of different symmetries and/or configurations should also be tested and analyzed in order to quantify and compare their effectiveness when applied to visual optics. Three types of wave-aberration sensors were developed and tested here. Each sensor has a very different configuration and/or symmetry (dodecagonal (DOD), cylindrical (CYL) and conventional Hartmann-Shack (HS)). METHODS: All sensors were designed and developed in the Physics Department of the Universidade de Sao Paulo--Sao Carlos. Each sensor was mounted on a laboratory optical bench used in a previous study. A commercial mechanical eye was used as control. This mechanical eye has a rotating mechanism that allows the retinal plane to be positioned at different axial distances. Ten different defocus aberrations were generated: 5 cases of myopia from -1D to -5D and 5 cases of hyperopia, from +1D to +5D, in steps of 1D following the scale printed on the mechanical eye. For each wavefront sensor a specific image-processing and fitting algorithm was implemented. For all three cases, the wavefront information was fit using the first 36 VSIA standard Zernike polynomials. Results for the mechanical eye were also compared to the absolute Zernike surface generated from coefficients associated with the theoretical sphere-cylinder aberration value. RESULTS: Precision was analyzed using two different methods: first, a theoretical approach was used by generating synthetic Zernike coefficients from the known sphere cylinder aberrations, simply by applying sphere-cylinder equations in the backward direction. Then comparisons were made of these coefficients with the ones obtained in practice. Results for DOD, HS and CYL sensors were, respectively, as follows: mean of root mean square (RMSE) for all aberrations, when theoretical Zernike coefficients were used as control, was 0.22, 0.66 and 0.26 microns; RMSE of sphere-cylinder values when compared to autorefractor measurements was 0.18D, 0.22D and 0.35D for sphere, 0.14D, 0.24D and 0.17D for cylinder, 34.36 degrees, 35.16 degrees and 26.36 degrees for axis; RMSE of sphere cylinder values when theoretical values were used as control was 0.11D, 0.29D and 0.46D for sphere, 0.15D, 0.28D and 0.17D for cylinder, 19.71 degrees, 25.56 degrees and 18.56 degrees for axis. CONCLUSION: The main conclusion is that the symmetry of an optical sensor is not an important consideration when measuring typical eye aberrations such as defocus (myopic and hyperopic), but there are differences. In this sense, the polar symmetry sensors render results that are equivalent to the traditional Cartesian Hartmann-Shack sensor, but furnish an easier method for determining the optical center. PMID- 16699678 TI - Unilateral blindness as a complication of nasal septoplasty: case report. AB - Blindness as a complication of nasal septoplasty is a devastating occurrence with only a few cases reported in the literature. In most of these cases, visual loss was attributed to retinal embolism following intra-arterial injection of substances into the nasal turbinate. This paper documents one case of complete unilateral blindness from direct optic nerve trauma following an apparently uneventful septorhinoplasty. Ophthalmologists and otolaryngologists should therefore be aware of the possible occurrence of such complication. PMID- 16699679 TI - Retinal nerve fiber layer loss documented by Stratus OCT in patients with pituitary adenoma: case report. AB - PURPOSE: To report abnormalities of retinal nerve fiber layer (RNFL) thickness using optical coherence tomography (Stratus OCT) in patients with pituitary adenoma. METHODS: Two patients with long-standing bitemporal visual field defects and optic nerve band atrophy were submitted to optical coherence tomography examination (Stratus OCT). RESULTS: Both patients with band atrophy revealed diffuse loss of the retinal nerve fiber layer on Stratus OCT, with severe reduction in the nasal and temporal areas of the optic disc. Retinal nerve fiber layer loss correlated well with visual field loss and with previous histological studies of band atrophy of the optic nerve. CONCLUSIONS: Stratus optical coherence tomography can provide useful information in the diagnosis of band atrophy from chiasmal lesions such as pituitary adenomas. PMID- 16699680 TI - [Retinopathy in a patient with hepatitis C treated with pegylated interferon and ribavirin: case report]. AB - Interferon is an immunomodulating cytokine used to treat patients with different diseases, such as hepatitis C chronic infection. Pegylated interferon is a new type of interferon, developed to increase the half-life of the drug. Many side effects have been related to its use, including ocular toxicity and retinopathy. The most reported ocular findings are cotton-wool spots and hemorrhages located at the posterior pole and surrounding optic nerve head. We describe one case of pegylated interferon-associated retinopathy with visual loss. The patient had visual acuity improvement four weeks after discontinuation of the medication and the ocular findings became much more subtle. PMID- 16699681 TI - [LASIK for high anisometropia in adults: case report]. AB - PURPOSE: To evaluate the efficacy of the laser in situ keratomileusis (LASIK) technique in the reduction of anisometropy in adults, for whom conventional treatment was not successful. METHODS: A sequence of cases of three eyes of three adult patients, two females and one male, age ranging from 28 and 49 years (average 38.3 years), who underwent LASIK. Two patients were followed up for eighteen months and one for six months. RESULTS: Comparing the corrected visual acuity before surgery with the non-corrected visual acuity after surgery, one eye gained two lines of vision, one eye remained the same and one eye lost one line of vision. All eyes remained without spheric refraction, and astigmatism did not exceed -0.75 D after surgery. There has been an improvement in the symptoms of asthenopia in the three patients. CONCLUSION: The LASIK technique proved to be efficient in the correction of anisometropy in adults, improving visual accuity and asthenopia complaints, and leading to a similar refraction in both eyes. PMID- 16699682 TI - [Palpebral cryptococcosis: case report]. AB - This paper is about a patient with acquired immunodeficiency syndrome empirically treated for miliary tuberculosis. During the clinical evolution the patient presented lesions compromising the right eyelid and tarsal conjunctiva. The initial diagnostic hypothesis was ocular tuberculosis with conjunctival and eyelid involvement. The biopsy of the conjunctival lesion identified an encapsulated yeast-like fungus: Criptococcus neoformans. After starting treatment with B anfotericin, the cutaneus lesions cleared. PMID- 16699683 TI - [Vitelliform macular detachment associated with basal laminar drusen: case report]. AB - The vitelliform macular lesion is usually observed in Best's disease and the adult type of foveomacular dystrophy. We describe a rare case of macular vitelliform lesion in association with basal laminar drusen. The patient has been followed at the Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo for the last four years. The differential diagnosis, clinical evolution, pathogenesis and treatment of this rare condition are discussed. PMID- 16699686 TI - Investments in health research: terms of reference for scientific and technological development in Brazil. PMID- 16699684 TI - [Association of human histocompatibility antigens with ophthalmological disorders]. AB - Many studies have been trying to identify genetic markers for ophthalmological diseases, including, among others, the HLA (Human Leukocyte Antigens). Localized on the short arm of chromosome 6, the human leukocyte antigen system is well known for its capacity to confer susceptibility or resistance to different diseases. In view of its accentuated polymorphism, the strength and type of association differs with the disease and sometimes, with the studied ethnic racial group. The development of molecular methods to typify HLA alleles and recent updates of their nomenclature has contributed to a better understanding of this system. In this review, some aspects of the human leukocyte antigen system are discussed, such as the methods of detection, nomenclature and association with acute anterior uveitis, ocular cicatricial pemphigoid, young-onset keratoconus and birdshot retinochoroidopathy. PMID- 16699688 TI - Opinions of the staff and users about the quality of the mental health care delivered at a Family Health Program. AB - Authors aimed at learning about the mental health care provided at a Family Health Program. 142 subjects were interviewed (18 nurses, 78 agents, 17 patients and 29 family members). In order to collect data, authors used the Scale of Opinion on Mental Health Care with 25 statements on the care, orientation by the members of the Program of Family Health team and the benefits of this Program. The total scores showed a difference between the agents and other groups. Among nurses, they found the shorter deviation and among the patients the greater variability in the responses. They emphasize the need for efficacy in arranging the appointments. Authors found a lack of coherence among the opinions of the subjects regarding the professionals' orientations. PMID- 16699687 TI - [Drug consumption and occupational violence in working women, a multicenter study: Mexico, Peru, Brazil]. AB - The purposes of the study were: 1) Determine the proportion of working women who consume drugs; 2) identify some occupational and personal risk factors that can predict drugs consumption; 3) identify the presence of occupational violence and its relation with drugs consumption; 4) identify differences and similarities in drugs consumption and occupational violence among women from three communities in Mexico (Monterrey), Peru (Lima) and Brazil (Rio de Janeiro). A multicenter, descriptive, correlational and comparative study was carried out, with a sample of 903 women. The results show that 11% of the participants in Mexico consume alcohol, 53% in Peru and 45% in Brazil. The consumption of illicit drugs corresponded to 5% in Mexico and 6% in Peru. The presence of occupational violence was found in 16% of the Mexican participants, 24% of the Peruvians and 39% of the Brazilians. PMID- 16699689 TI - [Pap smear realization profile of women between 40 and 49 years registered at a family health center]. AB - This descriptive and exploratory research aims to draw the profile of women between 40 and 49 years at a Family Health Center in Ribeirao Preto, Brazil, in view of the following variables: registration at the service, health insurance and smear realization date, among others. Data were collected in the patient records of 213 women. Epi Info 6.0 was used for data input and analysis and database creation. Results display that 49% of the women realized the pap smear at least once in the period from 2000 to 2003. 54.5% of them did not have a health insurance plan and smear realization rates are higher in this group. It was concluded that the organization of health care in accordance with the Family Health model favors adhesion to this preventive test. We recommend that new studies be carried out to obtain a more representative profile of the population that is registered at the service. PMID- 16699691 TI - [Changes in social roles: a consequence of traumatic brain injury for the family caregiver]. AB - The purpose was to study changes in the caregiver's social roles after traumatic brain injury (TBI), relating them with the degree of importance of these roles and with the condition of the victim six months or more after the TBI. The research was developed at the Brain Trauma Clinic of the University of Sao Paulo Hospital das Clinicas with 50 caregivers and 50 victims of TBI, by means of interviews and patient file analysis. A checklist was used for the interview with the caregiver, in order to identify the changes and importance of their social roles. The roles that had suffered most interruption due to the trauma were: friend, amateur/entertainment, family member and worker. The role of being a caregiver was the one that presented more modification. No association was found between change of role and the variables: condition of victim after TBI and importance of the social roles for the caregiver. PMID- 16699690 TI - [Health care in groups from the perspective of the elderly]. AB - This study aimed to describe and analyze the activities in elderly groups at basic health care units. Suffering from chronic-degenerative diseases, the elderly need continuous and more frequent health care. This can be facilitated by group actions, privileging educational activities. We found great diversity in methods, responsible teams and group composition. As to the patients, this activity allows for a better understanding and acceptance of the illness, as well as for the recovery of social roles and the occupation of leisure time. For services, the groups permit increased bonding. However, this requires training and permanent education for professionals, with a view to a significant dialogue. The increasing tendency of these actions in Campinas/SP is accompanying the guidelines of the Family Health PAIDEIA project. The group is a possible and privileged space to apply the PAIDEIA method. PMID- 16699692 TI - [Nurses' conceptualization on childbirth preparation]. AB - Descriptive, exploratory and qualitative study, carried out through the Delphi Technique, with a sample of 32 obstetrics specialist nurses working in 3 hospitals and 9 primary health care centers in the district of Oporto, Northern Portugal. The purposes of this work were: 1. to identify a group of nurses' ideas and consensual conceptions about Childbirth Preparation; 2. to know which information sources are consensually more used by nurses; 3. to know how they accept Childbirth Preparation programs as a way for pregnant women to have an active delivery. Results revealed that Childbirth Preparation is, according to the participants, an educational moment toward health, involving technical, educational, relational and informative procedures of great importance to pregnant women and nurses. This involves the period from the start of pregnancy to the puerperium, as a means of changing wrong behaviors with a view to achieving an expected goal of health gains for pregnant women and their family. PMID- 16699694 TI - [Sharing the death process with the family: a nurse's experience in the pediatric ICU]. AB - Death is present in the daily reality of nurses who work with children in ICU. This research aimed to comprehend nurses' experience while taking care of children and his/her family experiencing the death process. Symbolic Interactionism was adopted as a theoretical reference framework, while Interpretative Interactionism was used as a methodological reference framework to analyze the biographical narratives of seven nurses who took part in the research. The events that determined the nurses' history in this context were: FACING THE DEATH OF THE CHILD, PROJECTING YOURSELF IN THE MOTHER'S ROLE and PROMOTING FAREWELL. The analyzed data made it possible to understand the nurses' experience in care for children and their families during the death process. PMID- 16699693 TI - [Adolescent pregnancy from a family perspective: sharing projects of life and care]. AB - This qualitative study aimed to apprehend the meaning of adolescents' pregnancy for their families, using semistructured interviews and collective subject discourse. Adolescent pregnancy is represented as a problem to be faced with family support. The families worry and are mobilized to solve adversities. Besides the shock about the news, impotence as to pregnancy prevention, conformism, happiness and improvement in family relationships due to the baby's arrival, participants evidenced frustration due to the interruption/change in the family life project in terms of the adolescent being pregnant without a stable relationship with the child's father. In valuing the family perspective on adolescent pregnancy, professional care to pregnant adolescents and their families can be delivered in partnership with the family and social context, making it easier to cope with conflicts and recognizing the family as an active subject in this process. PMID- 16699695 TI - Chemical occupational risks identified by nurses in a hospital environment. AB - Hospital nursing workers are exposed to occupational chemical risks. This quantitative study aimed to identify what chemical substances nurses have contact with in their activities, what substances cause health problems and what alterations correspond to possible problems caused by the chemical products they mention. A self-administered data collection instrument was answered by 53 nurses, who mentioned exposure mainly to antibiotics and benzene (100%), iodine (98.1%) and latex-talc (88.7%); the main problem-causing substances mentioned were antineoplastic substances (86.7%), glutaraldehyde (79.2%) and ethylene oxide (75.5%); the described health alterations were: eye watering; allergic reactions; nausea and vomiting, while other health problems that can be caused by the above listed products were not mentioned. These workers need further information on occupational chemical risks, which they could have received in undergraduate or permanent education courses. PMID- 16699696 TI - [Visually impaired teenagers: perceptions on sexuality]. AB - This exploratory-descriptive study aimed to reveal perceptions of visually impaired teenagers on their sexuality. Data was collected from May to June 2004 by means of semistructured interviews, carried out with eight visually impaired teenagers studying at an institution in Porto Alegre, Brazil. The collected information was analyzed through content analysis according to Bardin. Two main categories emerged: Sexuality and Socio-behavioral changes. The research participants lack information on various sexuality-related issues, such as: knowledge on physiology, anatomy, affection and preventive care. Health professionals, especially nurses, need to be prepared to deal with these issues and to contribute to the development of a healthy sexuality of these individuals. PMID- 16699698 TI - [Understanding the gap between practice and the technical-scientific evolution of the bed-bath]. AB - The objectives of this qualitative study were to understand the experience of the nursing dependent person for the bed-bath and develop a theoretical model of this experience. The Symbolic Interactionism perspective and Grounded Theory methodology were used to develop the study. Observation and interview were the strategies used for data collection. Based on data analysis, a phenomenon was identified: learning to assess life and nursing when the patient becomes dependent on nursing care for the bed-bath. The theoretical model shows the meaning of the patient experience in the bed-bath, integrated by the core category--proposing a nursing model for the bed-bath according to the patient assessment process, as opposed to the technical-scientific evolution of this procedure. PMID- 16699697 TI - The sexuality of patients with onco-hematological diseases. AB - We interviewed 20 patients staying at a hospital unit, by means of a data collection instrument that is based on the eclectic model, with a view to characterizing the biological, psychological and sociocultural aspects involving human sexuality which are affected in patients with onco-hematological diseases. The research complied with ethical requirements for studies involving human beings. The results revealed that these clients presented problems related to biological aspects, mainly with respect to the phase of sexual desire (60% of the sample), sexual excitation (75%) and orgasm (75%). The psychological aspects related to sexual self-image were affected in 60% of the sample; problems related to social aspects (85%) were mainly due to the fear of acquiring an infection as a result of the low immunity provoked by the disease and treatment. These clients demonstrated alterations in their sexual function and in the way they expressed their sexuality. PMID- 16699699 TI - [Nursing physical examination of the full-term neonate: self-instructional software]. AB - The purpose of this research is to elaborate software about the physical examination of full-term newborns (TNB) for neonatal nursing teaching at undergraduate level. The software was developed according to the phases of planning, content development and evaluation. The construction of the modules was based on Gagne's modern learning theory and structured on the Keller Plan, in line with the systemic approach. The objectives were to elaborate and evaluate the contents of the self-instructional modules, to be used as a teaching strategy in the undergraduate course. After being structured, the material was reviewed and analyzed by 11 neonatal nursing experts, who rated the 42 exposed items as good or excellent. PMID- 16699700 TI - [Negative and positive eugenics: meanings and contradictions]. AB - Eugenics constitutes an important subject of debate, associated with current biogenetics improvements. Considering that the central point in eugenics has always been the preoccupation with future generations' health and constitution, and that the use of scientific means and knowledge for the birth of a physically and mentally healthy child can be considered a eugenic action, this paper tries to analyze the meanings and contradictions of negative and positive eugenics actions, constructed concomitantly with 20th-century technical-scientific improvements. The meanings range, respectively, between limiting or stimulating human reproduction, at the beginning of this century, and preventing diseases or improving physical and mental characteristics, nowadays. In the implantation of actions, contradictions were produced, such as the discrimination and elimination of many people in view of one ideal man, the biologization of eminently social factors, the defense of a supposed scientific neutrality and the indiscriminate use of the reproductive choice right. PMID- 16699701 TI - Historical evolution of the concept environment proposed in the Roy Adaptation Model. AB - The concern for the development of concepts becomes even more important when we report to its use in Nursing theories, in which some concepts are considered central, such as person, environment, health and nursing. This study aims to discuss the historical evolution of the environment concept used in the Roy Adaptation Model. The method of analysis the study was based on is the evolutionary conceptual analysis of the concept Environment of the Roy Adaptation Model. According to the research results, Environment is introduced as the second major concept in the Roy Adaptation Model. In this theorist's interpretation of the environment, we notice her interest in surpassing the mechanistic point of view upon which the first version of the model was based. The approach of the concept of interaction between people and groups seems to represent an improvement, influenced by the current thoughts that are preeminent in nursing. PMID- 16699702 TI - Writing for international publication in nursing journals: a personal perspective (part 1). AB - The number of printed and electronic (Internet) academic nursing publications in Brazil and around the world highlights the importance attached to publishing in the field of nursing. Internationally, journals are ranked according to their professional merits and peer review orientations. Financial institutions increasingly value publications in renowned journals as one criterion for granting funds for research. One important reason why many scientific articles do not meet the requirements from international journal reviewers, especially those submitted English, is the result of poor and literal translation of the text. The challenge we are facing in Latin America is to encourage the development of articles for publication in internationally reviewed journals. Co-authorship is a potentially stimulating model for researchers and postgraduate students to publish. This task can be undertaken through the help of international supervisors and researchers, supervisors or postgraduate students with good command of the English language. This article aims to demystify the publication process and present some guidelines on how to publish in international journals. PMID- 16699703 TI - [Pain measurement in the elderly: a review]. AB - This study revised and examined the theoretical and practical base of pain measurement in elderly persons through clinical studies and bibliographical reviews. We discussed the advantages and limitations of the ordinal instruments more frequently used in situations in which alterations characteristic of aging coexist, whose actions in feelings of pain have not been clarified yet. Moreover, we indicated the factors that intervene in the evaluation process of painful experiences leading to underestimation and inadequate control of pain in elderly persons. PMID- 16699704 TI - [Humanization in health care]. AB - This study aims to reflect on humanization in health care, recovering the history of understanding about mankind, the human and humanity, until humanization in humanity and health. We discuss the national humanization program in hospital care and reflect on this proposal and on the issue of humanization in Brazilian health care nowadays. Communication is indispensable to establish humanization, as well as technical and material conditions. Both users and health professionals need to be heard, building a network of dialogues to think and promote singular humanization actions. For this process to take effect, there is a need to involve the whole that makes up the health service. This group involves different professionals, such as managers, public policy makers, professional councils and education institutions. PMID- 16699705 TI - [Reflections on nursing teaching and students' first contact with the profession]. AB - Nursing teaching has been characterized by constant discussions on pedagogical proposals and implementation of curricular changes. The aim of this study was to reflect on nursing students' experience related to their first contacts with the profession, considering traditional perspectives and current tendencies that imprint values and attitudes in learning and practice. Nowadays, the complexity of human beings and the environment they live in is considered within a biopsychosocial, economic and cultural context, constituting resources that allow for the formation of competent professionals in order to deal with the challenges of the XXIst century. Thus, nursing schools have to follow a teaching philosophy to form trained people to attend to local and regional demands and are socially committed to change. What is aimed for is participative learning, in which teacher and student, articulated in the knowledge improvement process, do not forget about humanitarian views. PMID- 16699707 TI - Epidemiology of visceral leishmaniasis through spatial analysis, in Belo Horizonte municipality, state of Minas Gerais, Brazil. AB - The geographic information system approach has permitted integration between demographic, socio-economic and environmental data, providing correlation between information from several data banks. In the current work, occurrence of human and canine visceral leishmaniases and insect vectors (Lutzomyia longipalpis) as well as biogeographic information related to 9 areas that comprise the city of Belo Horizonte, Brazil, between April 2001 and March 2002 were correlated and georeferenced. By using this technique it was possible to define concentration loci of canine leishmaniasis in the following regions: East; Northeast; Northwest; West; and Venda Nova. However, as for human leishmaniasis, it was not possible to perform the same analysis. Data analysis has also shown that 84.2% of the human leishmaniasis cases were related with canine leishmaniasis cases. Concerning biogeographic (altitude, area of vegetation influence, hydrographic, and areas of poverty) analysis, only altitude showed to influence emergence of leishmaniasis cases. A number of 4673 canine leishmaniasis cases and 64 human leishmaniasis cases were georeferenced, of which 67.5 and 71.9%, respectively, were living between 780 and 880 m above the sea level. At these same altitudes, a large number of phlebotomine sand flies were collected. Therefore, we suggest control measures for leishmaniasis in the city of Belo Horizonte, giving priority to canine leishmaniasis foci and regions at altitudes between 780 and 880 m. PMID- 16699708 TI - Genetic structure of Triatoma venosa (Hemiptera: Reduviidae): molecular and morphometric evidence. AB - Triatoma venosa presents a restricted geographical distribution in America and is considered as a secondary vector of Chagas disease in Colombia and Ecuador. A total of 120 adult insects were collected in domestic and peridomestic habitats in an endemic area of the department of Boyaca, Colombia, in order to determine their genetic structure through morphometric and molecular techniques. The head and wings of each specimen were used for the analyses of size, shape, and sexual dimorphism. A significant sexual dimorphism was found, although no differences in size among the studied groups were detected. Differences were found in the analyzed structures except for male heads. DNA was extracted from the legs in order to carry out the internal transcriber space-2 (ITS-2) amplification and the randon amplified polymorphic DNA (RAPD) analyses. Length polymorphisms were not detected in the ITS-2. Fst and Nm values were estimated (0.047 and 3.4, respectively). The high genetic flow found among the insects captured in the domicile and peridomiciliary environment does not permit a genetic differentiation, thus establishing the peridomicile as an important place for epidemiological surveillance. PMID- 16699709 TI - Variation in the hatching response of Ochlerotatus albifasciatus egg batches (Diptera: Culicidae) in temperate Argentina. AB - Egg hatching of winter-collected Ochlerotatus albifasciatus was studied for six months. Batches of eggs were divided into two groups, one of them was stored in the laboratory at 23 degrees C and 12:12 photoperiod, and the other in the field under dead leaves. Every month, from July to December, eggs from the two groups were flooded under both laboratory and field conditions. Unhatched eggs were returned to the original condition and flooded two more times separated by ten day intervals. Results show that egg diapause is expressed in different intensities, not only on eggs exposed to different conditions but also in those exposed to the same condition, even when they were laid by the same female. Successive inundations yielded incomplete hatches of eggs, and favored the hatching response in the next flooding. Low environmental temperatures before and during the flooding depressed hatching response. This shows that eggs need a warm period before flooding as well as warm temperatures during flooding, to hatch. As drought period was longer hatching response increased, but this was also accompanied by warmer environmental conditions. The experiment performed in laboratory did not show that increment. Field studies showed that a layer of dead leaves protected eggs from extreme temperatures. PMID- 16699711 TI - Genetic characterization of St. Louis encephalitis virus isolated from human in Sao Paulo, Brazil. AB - The molecular characterization of SPH253157, a new strain of St. Louis encephalitis virus (SLEV), isolated in 2004 from the first case of human infection recognized in the state of Sao Paulo, Brazil, is reported. The patient, presenting a febrile illness without neurological involvement, was hospitalized as a probable case of dengue fever. Genomic RNA was isolated from the supernatant of C6/36 cells infected with acute phase-serum specimen of the patient and the envelope gene was amplified by reverse-transcription-polymerase chain reaction. The complete nucleotide sequence of the envelope gene of this isolate was directly sequenced from the amplified products and compared with other Brazilian and American SLEV strains. Phylogenetic analyses were carried out under maximum likelihood criterion with outgroups both included and excluded. Outgroups comprised four flavivirus of the Japanese encephalitis group. Phylogeny also included Bayesian analysis. The results indicated that the new SLEV isolate belongs to lineage III, being closely related to an Argentinean strain recovered from Culex sp. in 1979. It is concluded that there are at least 3 lineages of SLEV in Brazil. PMID- 16699710 TI - The fumigant and repellent activity of aliphatic lactones against Pediculus humanus capitis (Anoplura: Pediculidae). AB - New alternative insecticides are necessary for the chemical control of head lice. In this study the fumigant knockdown time 50% (KT50) and repellency index (RI) of three aliphatic lactones was compared with two essential oils and DDVP, against permethrin-resistance Pediculus humanus capitis from Argentina. In the fumigant assay, none of the lactones were effective compared to the highest activity of eucalyptus (KT50 15.53 m). In the repellency test, the three lactones were equally or more effective (RI ranging from 60.50 to 76.68) than the positive control (piperonal). These lactones are promising as head lice repellents. PMID- 16699712 TI - Distribution and ecological aspects of Rhodnius pallescens in Costa Rica and Nicaragua and their epidemiological implications. AB - In light of the Central American Initiative for the control of Chagas disease, efforts were made on the part of Costa Rican and Nicaraguan teams, working separately, to determine the present status of Rhodnius pallescens in areas close to the common border of the two countries, where the insect has appeared within the last few years. The opportunity was also used to establish whether R. prolixus, a vector present in some areas of Nicaragua, has been introduced in recent years into Costa Rica with Nicaraguan immigrants. It became evident that wild adults of R. pallescens are common visitors to houses in different towns of a wide area characterized as a humid, warm lowland, on both sides of the frontier. Up to the present, this bug has been able to colonize a small proportion of human dwellings only on the Nicaraguan side. There was strong evidence that the visitation of the adult bug to houses is related to the attraction of this species to electric lights. There were no indications of the presence of R. prolixus either in Nicaragua or in Costa Rica in this area of the Caribbean basin. Triatoma dimidiata, a widespread domestic species in both countries, was totally absent in the explored areas of Costa Rica but occasionally occurs on the Nicaraguan side. Serological surveys in children of both areas showed that transmission of Chagas disease takes place in a rather small degree in Costa Rica and more commonly in Nicaragua, indicating that R. pallescens could be a potential threat as a vector in this particular region. PMID- 16699713 TI - Emergence of dhfrXVb and blaCARB-4 gene cassettes in class 1 integrons from clinical Pseudomonas aeruginosa isolated in Amazon region. AB - Integrons play a role in horizontal acquisition and expression of genes, as well as gene reservoir, contributing for the resistance phenotype, particularly relevant to bacteria of clinical importance. We aimed to determine the composition and the organization of the class 1 integron variable region present in Pseudomonas aeruginosa clinical isolates from Brazil. Strains carrying class 1 integrons were resistant to the majority of antibiotics tested, except to imipenem and ceftazidime. Sequence analysis of the integron variable region revealed the presence of the blaCARB-4 gene into two distinct cassette arrays: aacA4-dhfrXVb-blaCARB-4 and aadB-aacA4-blaCARB-4. dhfrXVb gene cassette, which is rare in Brazil and in P. aeruginosa species, was found in one isolate. PFGE analysis showed the spread of blaCARB-4 among P. aeruginosa clones. The occurrence of blaCARB-4 and dhfrXVb in Brazil may contribute for developing resistance to clinically important antibiotics, and shows a diversified scenarium of these elements occurring in Amazon clinical settings, where no study about integron dynamics was performed to date. PMID- 16699714 TI - Ingestion of saliva during carbohydrate feeding by Lutzomyia longipalpis (Diptera; Psychodidae). AB - The aim of this study was to obtain experimental evidence that phlebotomine saliva is actually ingested during the carbohydrate ingestion phase (before and after blood digestion). The ingestion of carbohydrate was simulated as it occurs in the field by offering the insects balls of cotton soaked in sucrose, sucrose crystals or orange juice cells. The results obtained here showed that ingestion occurred under each condition investigated, as indicated by the presence of apyrase, an enzyme used as a marker to detect saliva in the insect gut and/or carbohydrate sources. Saliva ingestion by phlebotomine during the carbohydrate ingestion phase is important to explain how it could promote starch digestion and to trigger Leishmania promastigotes to follow a differentiation pathway as proposed previously by some authors. PMID- 16699715 TI - Toxicity of pyrethroids and repellency of diethyltoluamide in two deltamethrin resistant colonies of Triatoma infestans Klug, 1834 (Hemiptera: Reduviidae). AB - The aim of the currrent investigation was to evaluate (a) the toxicity of three pyrethroids (deltamethrin, lambda-cyhalothrin, and tetramethrin); (b) the effect of these insecticides on the locomotor activity; and (c) the repellent effect of N,N-diethyl-m-toluamide (DEET) on two deltamethrin-resistant strains of Triatoma infestans from Argentina (El Chorro and La Toma), and one susceptible strain. The resistance ratios (RRs) obtained for the La Toma strain were: > 10,769, 50.7, and > 5.2 for deltamethrin, lambda-cyhalothrin, and tetramethrin respectively. The RRs for the El Chorro strain were: > 10,769, 85.8, and > 5.2 for deltamethrin, lambda-cyhalothrin, and tetramethrin respectively. The hyperactivity usually caused by the three pyrethroids was in both the deltamethrin-resistant strains compared to the susceptible reference strain. No differences were observed in the repellent effect of DEET between the three groups. These results indicate that the deltamethrin-resistant insects have a cross resistance to lambda-cyhalothrin and tetramethrin, and are also resistant to the first symptom of pyrethroid poisoning (hyperactivity). However, the sensorial process related to DEET repellency does not appear to be altered. PMID- 16699716 TI - Phenotypic characterization of three clinical isolates of Burkholderia pseudomallei in Ceara, Brazil. AB - Burkholderia pseudomallei, the causative agent of melioidosis was found in a small cluster of cases in Tejucuoca, Ceara, Brazil. Tests were carried out to determine its phenotypic characteristics: colony morphology on Ashdown agar and MacConkey agar, biochemical profile in conventional biochemical tests and API 20NE, arabinose assimilation and susceptibility testing by disk diffusion, comparing with data in the literature. This study confirms the presence of B. pseudomallei in Brazil and describes its characteristics. PMID- 16699717 TI - Serologic evidence of Leishmania infection in free-ranging wild and domestic canids around a Brazilian National Park. AB - Transmission of disease between wildlife, domestic animals, and humans is of great concern to conservation issues and public health. Here we report on the prevalence of anti-Leishmania sp. antibodies in 21 wild canids (7 Chrysocyon brachyurus, 12 Cerdocyon thous, and 2 Lycalopex vetulus) and 74 free domestic dogs (Canis familiaris) sampled around the Serra do Cipo National Park. In dogs, the apparent prevalence was 8.1% and in wild canids it was 19% (2 crab-eating foxes, C. thous, and 2 maned wolves, C. brachyurus). Management of the domestic dog population with evaluation of incidence changes in humans and wildlife, and enlightenment on the role of wild reservoirs are essential issues for future action and research. PMID- 16699718 TI - Identification of human T-cell lymphotropic virus infection in a semi-isolated Afro-Brazilian quilombo located in the Marajo Island (Para, Brazil). AB - Antibodies to human T-cell lymphotropic virus-1 and 2 (HTLV-1 and 2) were tested in 259 inhabitants (98 males and 161 females) of four villages of the Marajo Island (Para, Brazil) using enzyme immunoassays (ELISA and Western blot). Types and subtypes of HTLV were determined by nested polymerase chain reaction (PCR) targeting the pX, env and 5 LTR regions. HTLV-1 infection was detected in Santana do Arari (2.06%) and Ponta de Pedras (1%). HTLV-2 was detected only in Santana do Arari (1.06%). Sequencing of the 5 LTR region of HTLV-1 and the phylogenetic analysis identified the virus as a member of the Cosmopolitan Group, subgroup Transcontinental. Santana do Arari is an Afro-Brazilian community and the current results represent the first report of HTLV-1 infection in a mocambo located in the Brazilian Amazon region. PMID- 16699719 TI - Superoxide dismutase in Cryptococcus neoformans varieties gattii, grubi, and neoformans. AB - Some clear dissimilarities occur among the varieties of Cryptococcus neoformans but there are few studies about the differences among individual yeast antioxidant enzymes. The total superoxide dismutase (SOD) activities and the copper, zinc-depend SOD (Cu,ZnSOD) and manganese-dependent SOD (MnSOD) isoenzymes of five reference C. neoformans strains belonged to A, B, C, AD and D serotypes (Table I) and other nine C. neoformans isolates (Table II) were determined. There were significant differences (p < 0.01 and p < 0.05) in total SOD activity among the varietie gattii (serotype C) and the other varieties. Cu,ZnSOD showed difference (p < 0.05) between A and D serotypes. These results point out a variety and serotype-independent SOD activity in C. neoformans reference strains and the other isolates that were evaluated. PMID- 16699720 TI - Compatibility of Biomphalaria tenagophila with Schistosoma mansoni: a study of homologous plasma transference. AB - This study aims to investigate the importance of the serum factors present in the plasma of resistant Biomphalaria tenagophila snails, when transferred to susceptible conspecific. Susceptible B. tenagophila (CF) received plasma from resistant B. tenagophila (Taim), and both were later infected with Schistosoma mansoni. We noticed that the plasma transfer showed an increase on the resistance of susceptible snails of about 86% when compared to the non-immunized group (p < 0.001). PMID- 16699721 TI - Identification of sex pheromones of Lutzomyia longipalpis (Lutz & Neiva, 1912) populations from the state of Sao Paulo, Brazil. AB - In Brazil, four populations of Lutzomyia longipalpis each producing different sex pheromones are recognised. It has been suggested that these chemotype populations represent true sibling species. In this study we present the results of an analysis, by coupled gas chromatography-mass spectrometry, of the pheromones of males L. longipalpis from two different municipalities of the state of Sao Paulo. Our study showed that L. longipalpis from these two municipalities produced different sex pheromones from each other. This coupled with the remarkable difference between the epidemiological situation in Aracatuba and Espirito Santo do Pinhal, suggests that the (S)-9-methylgermacrene-B and cembrene-1 populations may have different vectorial capacities. PMID- 16699722 TI - Leishmania infantum versus Leishmania chagasi: do not forget the law of priority. PMID- 16699723 TI - A comparison study of sampling and analyzing volatile organic compounds in air in Kuwait by using Tedlar bags/canisters and GC-MS with a cryogenic trap. AB - Kuwait experiences desert climatic weather. Due to the extreme hot and dry conditions in this country, some analytical phenomena have been discovered. Therefore, a systematic study of sampling and analyzing volatile organic compounds in air by using GC-MS with a cryogenic trap is reported in this paper. This study included comparisons of using different sample containers such as Tedlar bags and SUMMA canisters, and different cryogenic freezing-out air volumes in the trap. Calibration curves for different compounds and improvement of replicated analysis results were also reported here. The study found that using different sample containers produced different results. Analysis of ambient air samples collected in Tedlar bags obtained several volatile organic compounds with large concentrations compared to using SUMMA canisters. Therefore, to choose a sample container properly is a key element for successfully completing a project. Because GC-MS with a cryogenic trap often generates replicated results with poor agreement, an internal standard added to gas standards and air samples by using a gas syringe was tested. The study results proved that it helped to improve the replicated results. PMID- 16699725 TI - The -822G/A polymorphism in the promoter region of the MAP4K5 gene is associated with reduced risk of type 2 diabetes in Chinese Hans from Shanghai. AB - MAP4K5 (mitogen-activated protein kinase kinase kinase kinase 5), an early component of MAP kinase signal cascades was shown to activate Jun kinase in mammalian cells. The association between SNPs of MAP4K5 and type 2 diabetes (T2DM) was investigated due to the known relationship of the JNK pathway with T2DM. A total of 1,399 cases were included in the study. Oral glucose tolerance test (OGTT) and insulin release test (IRT) were performed, and blood DNA samples were extracted and genotyped on the MAP4K5 -822G/A site. These cases were subdivided into central-obesity and nonobesity groups, based upon their individual waist circumference. Allele-specific real-time PCR was employed for genotyping. No difference was found between the two groups in the distribution of three genotypes on MAP4K5 -822G/A. In the central-obesity group, fewer diabetic patients (38.9%) were present in the AA genotype group than the GG/GA group (58.5%, P=0.024). Glucose levels after 30 and 60 min of 75 g glucose tolerance, area under the curve for glucose, and insulin secretion indexes were lower (P<0.05) in AA than those in GG/GA genotype group in the central-obesity cases. Other variables did not show significant differences between the two groups. In the Han population from Shanghai, the AA genotype of MAP4K5 -822G/A in central obesity cases appears less likely to develop diabetes compared with the other genotypes. Therefore, the G allele may be a factor that does not protect central obesity cases from developing into diabetes. PMID- 16699726 TI - Benzo[a]pyrene-induced cell cycle progression is through ERKs/cyclin D1 pathway and requires the activation of JNKs and p38 mapk in human diploid lung fibroblasts. AB - Treatment of cells with carcinogen Benzo[a]pyrene (B[a]P) allows cells to evade G1 arrest and induces cells abnormal proliferation. However, the mechanisms of its action at cellular level are not well understood. To address this question, normal human embryo lung diploid fibroblasts (HELF) were selected in the present study. We found that exposure of cells with 2.5 microM of B[a]P for 24 h resulted in a decrease of G1 population by 11.9% (P < 0.05) and a increase of S population by 17.2% (P < 0.05). Treatment of cells with B[a]P also caused dose-related activation of MAPK and induction of cyclin D1 protein expression, whereas the CDK4 protein levels were not significantly affected by B[a]P. Overexpression of cyclin D1 protein stimulated by B[a]P was significantly inhibited by 50 microM AG126 (an inhibitor of ERK1/2), but not by 25 microM SP600125 (an inhibitor of JNK1/2) or 5 microM SB203580 (an inhibitor of p38 mapk), suggesting that B[a]P induced cyclin D1 expression was only regulated by ERK1/2 pathway. However, AG126, SP600125 or SB203580 led to cell cycle significantly arrested in G1 phase, indicating that ERK1/2, JNK1/2 and p38 mapk pathways are all required for B[a]P induced G1/S transition. In addition, HELF cells transfecting with antisense cyclin D1 cDNA or antisense CDK4 cDNA showed significantly G1 arrest after B[a]P stimulation. These results suggested that B[a]P exposure accelerated the G1-->S transition by activation of MAPK signaling pathways. Cyclin D1 and CDK4 are rate limiting regulators of the G1-->S transition and expression of cyclin D1 is predominantly regulated by ERK1/2 pathway in HELF cells. PMID- 16699727 TI - Kaempferol enhances endothelium-independent and dependent relaxation in the porcine coronary artery. AB - The vascular effects of kaempferol were investigated in isolated porcine coronary artery rings. U46619 (9,11-dideoxy-9alpha, 11alpha-methanoepoxy prostaglandin F2alpha, 30 nM) was used to contract porcine coronary arterial rings. Concentration relaxation curve of kaempferol (1 nM - 100 microM) was constructed and kaempferol demonstrated significant relaxation at high concentrations. At low concentration with no significant effect on relaxation, kaempferol (10 microM) enhanced relaxation produced by bradykinin, the calcium ionophore A23187, isoproterenol and sodium nitroprusside in endothelium-intact porcine coronary arteries. In endothelium-disrupt rings, kaempferol (10 microM) also enhanced the relaxation caused by isoproterenol, sodium nitroprusside, levcromakalim and nifedipine. On the other hand, antioxidant agents did not affect bradykinin induced relaxation or the enhancement effect of kaempferol. In summary, a low concentration of kaempferol (10 microM), devoid of significant vascular effect, has the ability to enhance endothelium-dependent and endothelium-independent relaxations. This action of kaempferol is unrelated to its antioxidant property. PMID- 16699736 TI - The population burden of fractures originates in women with osteopenia, not osteoporosis. AB - INTRODUCTION: Osteoporosis is associated with increased risk for fracture. However, most postmenopausal women have bone mineral density (BMD) within the normal or osteopenic range. The aim of this study was to determine the proportion of the population burden of fragility fractures arising from women at modest risk for fracture. METHODS: We measured baseline BMD in a population-based random sample of 616 postmenopausal women aged 60-94 years and followed these individuals for a median of 5.6 years (IQR 3.9-6.5) to determine the incidence of fractures according to age, BMD and the presence of a prior fracture. RESULTS: Based on WHO criteria, 37.6% of the women had normal total hip BMD, 48.0% had osteopenia and 14.5% had osteoporosis. The incidence of fracture during follow-up was highest in women with osteoporosis, but only 26.9% of all fractures arose from this group; 73.1% occurred in women without osteoporosis (56.5% in women with osteopenia, 16.6% in women with normal BMD). Decreasing BMD, increasing age and prior fracture contributed independently to increased fracture risk; in a multivariate model, the relative risk for fracture increased 65% for each SD decrease in BMD (RR=1.65, 95%CI 1.32-2.05), increased 3% for every year of age (RR=1.03, 95%CI 1.01-1.06) and doubled with prevalent fracture (RR=2.01, 95% CI 1.40-2.88). A prevalent fracture increased the risk for fractures such that women with osteopenia and prevalent fracture had the same, if not greater, risk as women with osteoporosis alone. CONCLUSIONS: Reducing the population burden of fractures requires attention to women with osteopenia, as well as osteoporosis, because over half of the fragility fractures in the population arise in these individuals, and women with osteopenia plus a prevalent fracture have the same fracture risk as women with osteoporosis. PMID- 16699737 TI - Correlations between bone mineral density and demographic, lifestyle, and biochemical variables in community-dwelling Japanese women 69 years of age and over. AB - INTRODUCTION: A few epidemiologic studies have comprehensively attempted to identify risk factors for low bone mineral density (BMD) in elderly Asian women. The purpose of this study was to identify demographic, lifestyle, and biochemical factors correlated with BMD in elderly Japanese women 69 years of age and over. METHODS: The study design was cross-sectional. The subjects were 583 ambulatory women aged 69 years and over, and their average age was 74.3 (SD 4.4) years. Predictor variables were age, reproductive history, anthropometric indices, grip strength, calcium intake, lifestyle information, and serum 25-hydroxyvitamin D, 1,25-dihydroxyvitamin D (1,25(OH)(2)D), osteocalcin (OC), and undercarboxylated osteocalcin (ucOC) values. The outcome variable was forearm BMD measured with a DTX-200 osteometer. RESULTS: Simple linear regression analyses showed that BMD was significantly positively associated with body height, weight, body mass index, grip strength, serum albumin concentration, and "housework," and negatively associated with age, years since menopause, age at menarche, number of children, serum 1,25(OH)(2)D concentration, serum OC concentration, and ucOC concentration. The stepwise multiple regression analysis showed that weight (beta=0.00316, SE=0.00028, R(2)=0.180), age (beta=-0.00321, SE=0.00050, R(2)=0.108), log-transformed serum OC (beta=-0.0445, SE=0.0064, R(2)=0.053), log transformed serum 1,25(OH)(2)D (beta=-0.0401, SE=0.0074, R(2)=0.050), "farmwork" (beta=0.00904, SE=0.00426, R(2)=0.005), and serum 25(OH)D concentration (beta=0.000281, SE=0.000120, R(2)=0.003) were significantly associated with BMD. CONCLUSION: It was concluded that body weight is a major predictor of forearm BMD among the factors measured in this study in independent Japanese women 69 years of age and over and that serum 1,25(OH)(2)D concentration may be associated with cortical BMD. Maintenance of body weight is very important for maintaining BMD in this population, unless a large weight aggravates obesity-related diseases. A follow-up study is needed to confirm these findings. PMID- 16699738 TI - Breast cancer and bone mass in older women: is bone density prescreening for mammography useful? AB - INTRODUCTION: The utility of screening mammography for older women with low bone mineral density (BMD) is controversial. This case-control study compares BMD at multiple sites in women with and without breast cancer to determine if BMD prescreening is useful in selecting women for continued screening mammograms. METHODS: Women diagnosed with breast cancer in the preceding 4 months and age matched controls (+/-2 years) with a normal mammogram, all aged 65 years and older, were recruited on a 1:2 basis; 237 women participated: 79 women (cases) with breast cancer and 158 controls. BMD at the lumbar spine, hip, radius, and whole body was measured with dual x-ray absorptiometry (DXA). RESULTS: Among women with breast cancer, 17.1% had stage 0, 41.5% stage I, 40.0% stage II, and 1.4% stage III. Women with breast cancer had larger waist circumferences (p=0.002) and waist-hip ratios (p=0.01), and they exercised less (p=0.002) than women of the control group. However, there were no differences between the cases and controls for age, obesity, and reproductive and menopausal history variables, or other covariates (p>0.10). There were no differences in lumbar spine, total hip, femoral neck, midshaft radius, or total body BMD (p>0.10), although the cases had higher BMD at the ultradistal radius than the controls (means: 0.527 vs. 0.516, respectively; p=0.014). There were no differences in breast cancer risk by tertile of BMD or osteoporosis status at the hip or spine. CONCLUSION: There is little difference in BMD between women with and without breast cancer. BMD is not useful as a prescreening predicator of mammography in older women and using it as such would result in cases of breast cancer being missed. PMID- 16699739 TI - Validation of various osteoporosis risk indices in elderly Chinese females in Singapore. AB - INTRODUCTION: This study aimed to compare the sensitivity and specificity of various published indices for identifying elderly Chinese females at risk of osteoporosis in Singapore. METHODS: The indices considered were the Simple Calculated Osteoporosis Risk Estimation (SCORE), the Osteoporosis Risk Assessment Instrument (ORAI), the Age Bulk One or Never Estrogens (ABONE), body weight (WEIGHT), and the Osteoporosis Self-Assessment Tool for Asians (OSTA). Altogether, 135 postmenopausal Chinese female subjects aged 55 years and older participated in the study, and their bone mineral density (BMD) was measured with dual-energy x-ray absorptiometry. Subjects were classified as osteoporotic if their femoral neck BMD T-score was -2.5 or lower. Receiver operating characteristic (ROC) curves were generated to determine the indices' cut-off points, sensitivity, and specificity. RESULTS: OSTA had the highest discriminatory power, with an estimated area under the ROC curve of 0.82. This was followed by SCORE (0.80), WEIGHT (0.78), ORAI (0.76), and ABONE (0.70). At the cut-off point of -2, OSTA achieved sensitivity and specificity of 91% and 59%, respectively. CONCLUSION: The study showed that OSTA is an effective index for identifying postmenopausal women at risk for osteoporosis. PMID- 16699740 TI - Technologies and solutions for data display in the operating room. AB - Recent advances in technology have led to the introduction of a variety of innovative devices, each with their own platform for data display, into the operating room (OR). While these innovative applications are expanding the traditional boundaries of the surgical space and enhancing treatment capabilities, the introduction of additional screens and displays is placing an ever-increasing load on the OR team. This review describes the main data display platforms currently available in ORs: computer monitors with CRT (cathode ray tube) or LCD (liquid crystal display) screens, suspended imaging displays, wearable computers (WC), auditory displays and tactile (haptic) displays. The different display platforms are evaluated according to their compatibility with the characteristics of the working environment (OR), the monitoring task, and the users (the surgical team). No single display configuration provides an ultimate solution for presenting patient data in the OR. A multi-sensory data display including visual, acoustic and haptic manipulation is suggested as a promising configuration for data display in the OR. PMID- 16699746 TI - Myocardial injury modulates the innate immune system and changes myocardial sensitivity. AB - OBJECTIVE: Transverse aortic constriction (TAC) results in a transient increase of proinflammatory cytokines, which return to baseline levels within 3 d. In contrast to cytokine baseline levels, the myocardium remains capable to respond even stronger to a new stimulus. As the molecular mechanisms for this phenomenon are unknown, we tested whether TAC modulates the innate immune system in mice and changes the inflammatory reaction to a new stimulus. METHODS: Following 3 d of TAC or sham-operation procedure (SOP), LPS (20 mg/kg) or PBS (control) were administered intraperitoneal for 10 min as well as for 6 h. Hemodynamic parameters were recorded to measure the effects of TAC and LPS. After TAC/SOP alone CD14 expression was monitored and after additional 6 h of LPS/PBS the expression of CD14, TLR4 and proinflammatory cytokines were determined by western blot, ELISA and RNase protection assay, respectively. Following TAC/SOP and 10 min of LPS/PBS, NFkappaB activation was investigated by EMSA. RESULTS: TAC induced cardiac hypertrophy and elevated blood pressure. LPS application led to hypotension and other symptoms of sepsis. CD14 expression increased after TAC alone and even further after additional LPS challenge. However, we did not detect changes of TLR4 expression. Also NFkappaB activation increased after LPS challenge higher in the TAC than in the SOP group. LPS-stimulation induced also higher cytokine expression in the TAC than in the SOP group. CONCLUSION: TAC modulates innate immunity by regulating the expression of CD14 and changes the myocardial tissue to respond more powerful to LPS. PMID- 16699748 TI - [The minimally invasive anterior approach to hip arthroplasty]. AB - A modified anterior approach to the hip developed by the senior author has been utilized in performing over 7000 hip replacement operations at this joint replacement center in the past three decades. It can be readily applied to both primary and revision surgery, and involves muscle splitting and sparing of the major hip muscles, innervation, and function. Since the late 1970s, Yale orthopaedic residents have been taught this minimally invasive anterior approach using one, two, or three mini-incisions without fluoroscopy, special operating room tables, or special retractors. The authors' overall experience has documented a very low incidence of dislocation and minimal perioperative complications. While a smaller skin incision is desirable by patients for cosmetic reasons and decreased healing time, what goes on beneath the skin is far more important. A well-placed total hip replacement should never be compromised. While many of these patients can ambulate the same or first postoperative day quite well, the authors do not advocate outpatient total hip arthroplasty as some proponents have. Bone is still cut, tissues bleed, clots can form, and the traditional complications of arthroplasty, while reduced, can still occur. PMID- 16699749 TI - The role of preoperative bone scan for determining the treatment method for femoral neck fracture. AB - To evaluate the role of preoperative bone scintigraphy in determining the operative treatment method for femoral neck fracture, we reviewed the data of 83 patients who underwent preoperative bone scanning after femoral neck fracture. Fractures were classified using the Garden staging system. Radioisotope uptake in femoral heads was evaluated visually. Of 28 patients with Garden stage I or II, radioactivity of the femoral head was normal in 26, partially reduced in one, and generally reduced in one patient. Twenty-seven patients were treated by closed reduction and multiple pinning, and one patient was treated by bipolar hemiarthroplasty. Of 55 patients with Garden stage III or IV, femoral-head radioactivity was normal in three, partially reduced in seven and generally reduced in 45 patients. Fifty-four patients were treated by bipolar hemiarthroplasty or total hip arthroplasty, and one patient was treated by closed reduction and multiple pinning. In only one of the 83 cases was the operative method changed because of bone scan findings. Isotope uptake of the femoral head after femoral neck fracture generally corresponded with the degree of fracture displacement. Preoperative bone scans appear to have no significant role to play in determining the operative treatment method for femoral neck fracture. PMID- 16699751 TI - Does 16-MDCT angiography scanning direction affect image quality of coronary artery bypass grafts and the native coronary arteries? AB - To assess the impact of scanning direction on the image quality of coronary artery bypass grafts (CABGs), native coronary arteries (NCAs) were examined by electrocardiographically (ECG) gated 16-row multidetector computed tomography (16 MDCT). Eighty-two patients with 209 grafts were studied by 16-MDCT. Forty-one patients with 111 grafts were scanned craniocaudally. Forty-one patients with 98 grafts were scanned caudocranially. CABG, native coronary arteries were examined in four (proximal, middle, distal, distal anastomoses), three (proximal, middle, distal) segments, respectively. Subjective image quality on a four-point scale was calculated for segments. Scores of groups were compared. Results Image quality scores of proximal, distal segments of the right coronary artery (RCA) were better in caudocranially scanned group (P<0.05). When we subgrouped patients according to initial heart rates (IHR) (group 1, <65 beats/min; group 2, > or =65 beats/min), there was no statistical significance between image quality scores of coronary arteries, CABG when IHR was <65 beats/min in groups regardless of scanning direction. Scores of anastomotic segment of CABG to RCA, middle segments of circumflex coronary artery, proximal and distal segments of RCA in caudocranially scanned group were better when the IHR is > or =65 beats/min compared with the craniocaudally scanned group. When the IHR of the patient is > or =65 beats/min, performing ECG-gated 16-MDCT angiography in the caudocranial direction provides better image quality for evaluation of coronary arteries and CABGs. PMID- 16699752 TI - Optimal image reconstruction intervals for non-invasive coronary angiography with 64-slice CT. AB - The reconstruction intervals providing best image quality for non-invasive coronary angiography with 64-slice computed tomography (CT) were evaluated. Contrast-enhanced, retrospectively electrocardiography (ECG)-gated 64-slice CT coronary angiography was performed in 80 patients (47 male, 33 female; mean age 62.1+/-10.6 years). Thirteen data sets were reconstructed in 5% increments from 20 to 80% of the R-R interval. Depending on the average heart rate during scanning, patients were grouped as < 65 bpm (n = 49) and > or = 65 bpm (n = 31). Two blinded and independent readers assessed the image quality of each coronary segment with a diameter > or = 1.5 mm using the following scores: 1, no motion artifacts; 2, minor artifacts; 3, moderate artifacts; 4, severe artifacts; and 5, not evaluative. The average heart rate was 63.3 +/- 13.1 bpm (range 38-102). Acceptable image quality (scores 1-3) was achieved in 99.1% of all coronary segments (1,162/1,172; mean image quality score 1.55 +/- 0.77) in the best reconstruction interval. Best image quality was found at 60% and 65% of the R-R interval for all patients and for each heart rate subgroup, whereas motion artifacts occurred significantly more often (P < 0.01) at other reconstruction intervals. At heart rates < 65 bpm, acceptable image quality was found in all coronary segments at 60%. At heart rates > or = 65 bpm, the whole coronary artery tree could be visualized with acceptable image quality in 87% (27/31) of the patients at 60%, while ten segments in four patients were rated as non-diagnostic (scores 4-5) at any reconstruction interval. In conclusion, 64-slice CT coronary angiography provides best overall image quality in mid-diastole. At heart rates < 65 bpm, diagnostic image quality of all coronary segments can be obtained at a single reconstruction interval of 60%. PMID- 16699755 TI - Effect of flocculation on performance of arming yeast in direct ethanol fermentation. AB - In the direct ethanol fermentation of raw starch by arming yeast with alpha amylase and glucoamylase, it is preferable to use a flocculent yeast because it can be recovered without centrifugation. Three types of arming yeast system, I (nonflocculent), II (mildly flocculent), and III (heavily flocculent), were constructed and their fermentation performances were compared. With an increase in the degree of flocculation, specific ethanol production rate for soluble starch decreased (0.19, 0.17, and 0.12 g g-dry-cell(-1) h(-1) for systems I, II, and III, respectively), but that for raw starch did not decrease as much as expected (0.06, 0.06, and 0.04 g g-dry-cell(-1) h(-1) for systems I, II and III, respectively). Microscopic observation revealed that many starch granules were captured in the yeast flocs in system III during the direct ethanol fermentation of raw starch. It was suggested that the capture of starch granules increases apparent substrate concentration for amylolytic enzymes in arming yeast cell flocs; thus, the specific ethanol production rate of system III was kept at a level comparable to those of the other systems. PMID- 16699756 TI - Marker and promoter effects on heterologous expression in Aspergillus nidulans. AB - To study the effects of selection marker, promoter type, and copy number on heterologous expression in Aspergillus nidulans, strains were constructed with single- and multicopy plasmid integrations bearing a reporter gene (lacZ) under the control of either an inducible (alcA) or constitutive (gpdA) promoter and one of three Aspergillus nutritional marker genes (argB, trpC, or niaD). beta Galactosidase activity in the transformants varied over three orders of magnitude, with the majority of levels in the range of 5x10(3)-1x10(4) U/mg. Significant differences in mean expression levels were found when comparing single-copy transformants with the same promoter but a different marker. Transformants with the argB marker had the highest average expression, approximately threefold over the trpC or niaD clones. For each promoter, maximal expression for the set was seen in the range of the single-copy clones, implying that increasing the copy number does not reliably increase expression in Aspergillus. PMID- 16699754 TI - Multiple cryptic splice sites can be activated by IDS point mutations generating misspliced transcripts. AB - Mutations in the gene encoding the enzyme iduronate-2-sulfatase (IDS) were reported as the cause of the X-linked recessive lysosomal disease, mucopolysaccharidosis II (MPS II). Amongst the different mutations, it emerges that nearly 10% are nucleotide substitutions causing splicing mutations. We now report the molecular characterisation of three MPS II patients with multiple aberrant transcripts due to three different point mutations. The c.418+1G>C that occurred in the invariant splice-site motif, produced only aberrantly spliced transcripts. Whilst the mutations affecting variant motifs (c.419G>T) or coding regions (c.245C>T) led to aberrantly spliced transcripts in addition to correctly spliced transcripts with the respective predicted missense mutation, p.G140V or p.A82V. A combination of experimental tests and computational approaches were used to understand the molecular basis underlying the altered transcription patterns. In addition, by using real-time reverse transcriptase polymerase chain reaction, the reduction of mRNA amount in two patients observed was likely due to nonsense-mediated mRNA decay pathway. Overall, our results further emphasised the importance of cloning and sequencing independent transcripts to reveal less abundant, aberrant products, which often could not be detected by direct sequencing. Moreover, the different splicing patterns observed in the three patients as a consequence of point mutations show how sensitive the balance is between constitutive and cryptic splice sites in the IDS gene. The generation of such diverse transcripts, together with their level of expression, could contribute to the profound phenotypic variability reported in MPS II. PMID- 16699757 TI - Construction of a large phage display antibody library by in vitro package and in vivo recombination. AB - Capacity and diversity are extremely important to the quality of various phage display libraries. In this work, lambda phage-based in vitro package was applied to construct a filamentous phage display antibody library so as to enlarge its capacity and introduce more sequence diversity in the final library. In vivo recombination via Cre recombinase/lox sites was also exploited to create V(H)/V(L) combination diversity based on multivalent package of lambda phage packaging extracts on phagemid DNA concatemers. The library constructed with 10 microg concatenated phagemid DNA and ten vials of lambda phage packaging extracts was calculated to contain 1.40 x 10(10) independent clones. Higher capacity can be easily achieved when more materials are consumed. This strategy is somewhat more efficient than prior methods. PMID- 16699758 TI - Anterior vertebral body replacement with a titanium implant of adjustable height: a prospective clinical study. AB - In the operative treatment of spinal injuries, the reconstruction of the anterior column of the thoracolumbar spine is still controversial. We conducted a prospective clinical study to investigate the clinical and radiological outcome of 50 patients treated with a vertebral body replacement of adjustable height (Synex). Fifty consecutive patients were evaluated during in-patient treatment and at 12 and 20 months post-operatively in clinical notes and radiographs. 38/50 patients were operated for traumatic fractures. Out of 50 patients 45 attended the follow-up clinic 1 year post-operatively and 39 of these patients were examined after 20 months. Twenty-five patients returned to pre-injury activities within 1 year. This number increased to 29/39 patients at 20 months. Seventy three percent of the patients returned to their job. After 1 year 25/45 patients complained of little or no back pain and 6 months later six patients were limited in their back function. At 1 year only three patients complained of surgical site pain which was improved at their final follow-up at 20 months. Individual satisfaction was determined using a score on a visual analog scale containing 19 questions on back pain, and functional limitation of the spine that has to be filled in by the patients at three different points of time. The score decreased from 87/100 pre-operatively to 65/100 at 1 year follow-up (P<0.001). The average permanent correction of the injured vertebra was 16.8 degrees (88%) including 2.3 degrees (12%) loss of correction at 12 months after operation. Bony integration was obtained in 83%. Early and intermediate outcome with the Synex vertebral replacement device for reconstruction of the anterior column appears promising. The loss of correction or reduction was only minimal. On the basis of our results we recommend the Synex implant as an alternative for the fixation and stabilisation of thoracolumbar fractures. However, long-term results and a clinically random control study are still required. PMID- 16699760 TI - Metabolism of acrylamide to glycidamide and their cytotoxicity in isolated rat hepatocytes: protective effects of GSH precursors. AB - Acrylamide (AA) is a widely studied industrial chemical that is neurotoxic, mutagenic to somatic and germ cells, and carcinogenic in rodents. The recent discovery of AA at ppm levels in a wide variety of commonly consumed foods has energized research efforts worldwide to define toxicity and prevention. Metabolism and cytotoxicity of AA and its epoxide glycidamide (GA) were studied in the hepatocytes freshly isolated from male Sprague-Dawley rats. The isolated hepatocytes metabolized AA to GA. The formation of GA followed Michaelis-Menten kinetic parameters yielded apparent Km = 0.477 +/- 0.100 and 0.263 +/- 0.016 mM, Vmax = 6.5 +/- 2.1 and 26.4 +/- 3.0 nmol/h/10(6) cells, and CLint = 14 +/- 5 and 100 +/- 12 microl/h/10(6) cells for the hepatocytes from untreated and acetone treated rats, respectively. There were lower Km and marked increases in Vmax (four-fold) and in CLint (sevenfold) in acetone-treated rat hepatocytes. The data suggest that CYP2E1 played a major role in metabolizing AA to more toxic GA. Both AA and GA induced a concentration- and time-dependent glutathione (GSH) depletion of the hepatocytes. From decreasing rates of GSH contents in hepatocytes, the parameters of glutathione S-transferase (GST) in hepatocytes to AA and GA were calculated to be Km = 1.4 and 1.5 mM, Vmax = 21 and 33 nmol/h/10(6) cells, and CLint = 15 and 23 microl/h/10(6) cells, respectively. GA 1.5-times more readily depleted GSH content than AA. GA decreased the viability of hepatocytes at 3 mM, but AA did not. These data indicate that GA is more toxic than AA as assessed by intracellular GSH depletion and loss of viability of hepatocytes. GSH precursors such as N-acetylcysteine and methionine provided significant anti-cytotoxic effects on the decrease of GSH content and cell viability of hepatocytes induced by GA and AA. PMID- 16699761 TI - [Clinical pathways: experiences, expectations, and perspectives]. AB - Due to economic necessities, existing reserves in clinical practice have to be used efficiently. The presentation of medical activity and systematic illustration of medical decisions in the form of clinical pathways constitute important tools to ensure the quality of patient care under fiscal pressure caused by the DRG system. To successfully establish clinical pathways, the architecture of the standard operating procedures developed specifically for a hospital must adequately reflect medical concerns and patient-related needs. The concept of necessity for the good of the patient applies in this instance rather than the principle of the complete depiction of all medical processes. Organizing the implementation into clinical practice should be assured and monitored. The standard operating procedures should be regularly adapted to suit medical developments. Success with the procedure should be reported and deviations from predetermined targets should be documented. A clinical pathway does not represent an administrative end in itself but rather a part of a higher control system that helps to improve the utilisation of the hospital's resources in terms of reliability and quality of patient care. PMID- 16699764 TI - Assessment of radiation dose awareness among pediatricians. AB - BACKGROUND: There is increasing awareness among pediatric radiologists of the potential risks associated with ionizing radiation in medical imaging. However, it is not known whether there has been a corresponding increase in awareness among pediatricians. OBJECTIVE: To establish the level of awareness among pediatricians of the recent publicity on radiation risks in children, knowledge of the relative doses of radiological investigations, current practice regarding parent/patient discussions, and the sources of educational input. MATERIALS AND METHODS: Multiple-choice survey. RESULTS: Of 220 respondents, 105 (48%) were aware of the 2001 American Journal of Roentgenology articles on pediatric CT and radiation, though only 6% were correct in their estimate of the quoted lifetime excess cancer risk associated with radiation doses equivalent to pediatric CT. A sustained or transient increase in parent questioning regarding radiation doses had been noticed by 31%. When estimating the effective doses of various pediatric radiological investigations in chest radiograph (CXR) equivalents, 87% of all responses (and 94% of CT estimates) were underestimates. Only 15% of respondents were familiar with the ALARA principle. Only 14% of pediatricians recalled any relevant formal teaching during their specialty training. The survey response rate was 40%. CONCLUSION: Awareness of radiation protection issues among pediatricians is generally low, with widespread underestimation of relative doses and risks. PMID- 16699765 TI - In vivo tumor angiogenesis imaging with site-specific labeled (99m)Tc-HYNIC-VEGF. AB - PURPOSE: We recently developed a cysteine-containing peptide tag (C-tag) that allows for site-specific modification of C-tag-containing fusion proteins with a bifunctional chelator, HYNIC (hydrazine nicotinamide)-maleimide. We then constructed and expressed C-tagged vascular endothelial growth factor (VEGF) and labeled it with HYNIC. We wished to test (99m)Tc-HYNIC-C-tagged VEGF ((99m)Tc HYNIC-VEGF) for the imaging of tumor vasculature before and after antiangiogenic (low continuous dosing, metronomic) and tumoricidal (high-dose) cyclophosphamide treatment. METHODS: HYNIC-maleimide was reacted with the two thiol groups of C tagged VEGF without any effect on biologic activity in vitro. (99m)Tc-HYNIC-VEGF was prepared using tin/tricine as an exchange reagent, and injected via the tail vein (200-300 microCi, 1-2 microg protein) followed by microSPECT imaging 1 h later. RESULTS: Sequencing analysis of HYNIC-containing peptides obtained after digestion confirmed the site-specific labeling of the two accessible thiol groups of C-tagged VEGF. Tumor vascularity was easily visualized with (99m)Tc/VEGF in Balb/c mice with 4T1 murine mammary carcinoma 10 days after implantation into the left axillary fat pad in controls (12.3+/-5.0 tumor/bkg, n=27) along with its decrease following treatment with high (150 mg/kg q.o.d. x 4; 1.14+/-0.48 tumor/bkg, n=9) or low (25 mg/kg q.d. x 7; 1.03+/-0.18 tumor/bkg, n=9) dose cyclophosphamide. Binding specificity was confirmed by observing a 75% decrease in tumor uptake of (99m)Tc/biotin-inactivated VEGF, as compared with (99m)Tc HYNIC-VEGF. CONCLUSION: (99m)Tc can be loaded onto C-tagged VEGF in a site specific fashion without reducing its bioactivity. (99m)Tc-HYNIC-VEGF can be rapidly prepared for the imaging of tumor vasculature and its response to different types of chemotherapy. PMID- 16699766 TI - Evaluation of D-isomers of O-18F-fluoromethyl, O-18F-fluoroethyl and O-18F fluoropropyl tyrosine as tumour imaging agents in mice. AB - The aim of this study was to evaluate the properties of the D-amino acid isomers O-(18)F-fluoromethyl tyrosine ((18)F-FMT), O-(18)F-fluoroethyl tyrosine ((18)F FET) and O-(18)F-fluoropropyl tyrosine ((18)F-FPT) as tumour-detecting agents with PET in comparison with the corresponding L-isomers. L- or D-(18)F-FMT, (18)F FET or (18)F-FPT, prepared by (18)F-fluoromethylation, (18)F-fluoroethylation or (18)F-fluoropropylation of L- and D-tyrosine, was intravenously injected into BALB/cA Jcl-nu mice bearing HeLa tumour cells. At 5, 15, 30 and 60 min post intravenous administration, the uptake of each compound in normal abdominal organs and xenotransplanted HeLa cells was determined using the tissue dissection method. Metabolic stability analyses of these compounds in the plasma were performed with the thin-layer chromatography method. In the plasma fraction, although L- and D-isomers of (18)F-FMT, (18)F-FET and (18)F-FPT provided comparable metabolic stability, D-isomers of these labelled compounds revealed a faster elimination rate than their L-isomers, with a higher peak uptake in the blood and kidney 5 min post administration. Compared with natural amino acid ligands, such as L-(11)C-methionine, the uptake of L-isomers of these labelled compounds was relatively low and stable in the abdominal organs, while D-isomers revealed much lower and faster clearance rates compared with the corresponding L isomers. Among the abdominal organs, the pancreas showed relatively high uptake of all the labelled compounds used here, and the uptake of D-isomers was much lower than that of the L-isomers. Although tumour uptake levels of D-isomers of (18)F-FMT, (18)F-FET and (18)F-FPT were almost 95%, 43% and 39% of the uptake levels of each of the L-isomers 60 min post administration, the tumour-to-blood ratios of these D-isomers were 181%, 137% and 101% of the ratios of the corresponding L-isomers. D-isomers of (18)F-FMT and (18)F-FET indicated improved tumour-to-liver ratios compared with the corresponding L-isomers, and D-(18)F-FPT showed the highest tumour-to-pancreas ratio among all the other compounds assayed here. These results suggest that D-isomers of (18)F-fluoroalkyl tyrosine analogues are potential tracers for tumour imaging with PET. PMID- 16699767 TI - Persistent aseptic meningitis due to post-surgical spinal CSF leakage: value of fused (111m)In-DTPA SPECT-CT cisternography. PMID- 16699768 TI - Simplified quantification of small animal [18F]FDG PET studies using a standard arterial input function. AB - PURPOSE: Arterial input function (AIF) measurement for quantification of small animal PET studies is technically challenging and limited by the small blood volume of small laboratory animals. The present study investigated the use of a standard arterial input function (SAIF) to simplify the experimental procedure. METHODS: Twelve [(18)F]fluorodeoxyglucose ([(18)F]FDG) PET studies accompanied by serial arterial blood sampling were acquired in seven male Sprague-Dawley rats under isoflurane anaesthesia without (every rat) and with additional (five rats) vibrissae stimulation. A leave-one-out procedure was employed to validate the use of a SAIF with individual scaling by one (1S) or two (2S) arterial blood samples. RESULTS: Automatic slow bolus infusion of [(18)F]FDG resulted in highly similar AIF in all rats. The average differences of the area under the curve of the measured AIF and the individually scaled SAIF were 0.11+/-4.26% and 0.04+/-2.61% for the 1S (6-min sample) and the 2S (4-min/43-min samples) approach, respectively. The average differences between the cerebral metabolic rates of glucose (CMR(glc)) calculated using the measured AIF and the scaled SAIF were 1.31+/-5.45% and 1.30+/-3.84% for the 1S and the 2S approach, respectively. CONCLUSION: The use of a SAIF scaled by one or (preferably) two arterial blood samples can serve as a valid substitute for individual AIF measurements to quantify [(18)F]FDG PET studies in rats. The SAIF approach minimises the loss of blood and should be ideally suited for longitudinal quantitative small animal [(18)F]FDG PET studies. PMID- 16699769 TI - Topography of cerebral monoamine transporter availability in families with SCA2 mutations: a voxel-wise [123I]beta-CIT SPECT analysis. AB - PURPOSE: The purpose of this study was to investigate the monoamine transporter status of dopamine, serotonin and norepinephrine throughout the brain in spinocerebellar ataxia type 2 (SCA2). To this end, nine patients were studied with [(123)I]beta-CIT SPECT. METHODS: Data were compared with ten age-matched healthy control subjects and ten patients with young-onset Parkinson's disease (YOPD), matched for age. Parametric SPECT images of the specific-to-non displaceable equilibrium partition coefficient (V (3)''), which is proportional to the receptor density (B (max)), were generated. In order to objectively localise focal changes in beta-CIT uptake throughout the brain volume without having to make an a priori hypothesis as to their location, statistical parametric mapping (SPM) was applied to SPECT images. Data clusters revealed by SPM, showing significant differences in V (3)'' values between groups, were transformed onto the individual V (3)'' image to obtain mean regional uptake values. RESULTS: Both SCA2 and YOPD patients showed significant decreases in striatal [(123)I]beta-CIT SPECT uptake when compared with controls. However, in SCA2 patients, additional reductions in caudate/anterior putamen, midbrain and pons [(123)I]beta-CIT uptake were localised with SPM. CONCLUSION: Voxel-wise analysis of [(123)I]beta-CIT SPECT revealed more widespread decline of monoamine transporter availability in SCA2 than in YOPD, reflecting differences in the underlying pathology. We suggest that the quantification of midbrain and pons [(123)I]beta-CIT signal is likely to improve the diagnostic accuracy in patients presenting with clinical features of both SCA2 and YOPD at initial investigation. PMID- 16699770 TI - Bilateral adrenal metastases from malignant melanoma: concordant findings on (18)F-FDG and (18)F-FDOPA PET. PMID- 16699771 TI - Sedation and analgesia in the PICU: many questions remain. PMID- 16699773 TI - [Hyperpigmentation of the abdominal skin. Diagnosis: Morbus Gougerot-Carteaud PCR]. PMID- 16699776 TI - Costs of chemotherapy in the treatment of colorectal cancer. PMID- 16699772 TI - Consensus guidelines on sedation and analgesia in critically ill children. AB - OBJECTIVE: The United Kingdom Paediatric Intensive Care Society Sedation, Analgesia and Neuromuscular Blockade Working Group is a multi-disciplinary expert panel created to produce consensus guidelines on sedation and analgesia in critically ill children and forward knowledge in these areas. Sedation and analgesia are recognised as important areas of critical care practice and adult clinical practice guidelines in these fields remain amongst the most popular of those produced by the Society of Critical Care Medicine. However, similar clinical practice guidelines have not previously been produced for the critically ill paediatric patient. DESIGN: A modified Delphi technique was used to allow the Working Group to anonymously consider draft recommendations in three Delphi rounds with predetermined levels of agreement. This process was supported by a total of four consensus conferences. Once consensus had been reached, a systematic review of the available literature was carried out. OUTCOME: A set of consensus guidelines was produced including 20 key recommendations, 10 relating to the provision of analgesia and 10 relating to the sedation of critically ill children. An evaluation of the existing literature supporting these recommendations is provided. CONCLUSIONS: Multi-disciplinary consensus guidelines for maintenance sedation and analgesia in critically ill children have been successfully produced and are supported by levels of evidence (excluding sedation and analgesia for procedures and excluding neonates). The working group has highlighted the paucity of high-quality evidence in these important clinical areas and this emphasises the need for further randomised clinical trials in this area. PMID- 16699777 TI - Ganglion cell containing pituitary adenomas: signs of neuronal differentiation in adenoma cells. AB - Ganglion cell containing pituitary adenomas are rare. They represent tumors originating in the sella turcica which are composed of adenomatous and neuronal components. Recently accumulated information suggests a common origin for their neuronal and pituitary constituents. The objective of this study was to report the clinical and morphologic findings of pituitary gangliocytomas and study their immunoprofile using neuronal markers. Seven cases of pituitary gangliocytomas retrieved from 1,322 sellar lesions were studied. All tumors were removed from patients with mild acromegaly. Histologically they were biphasic composed of pituitary adenoma and clusters of ganglion cells embedded in a variably dense neuropil substrate. All adenomas belonged to the category of sparsely granulated somatotroph adenoma and were positive for growth hormone, whereas in five tumors, a few adenoma cells were also positive for prolactin. Ganglion cells were immunoreactive for NSE, synaptophysin and neurofilament protein (NFP). NFP reactive fibrils were observed in the neuropil substrate and varied in number among the cases. Interestingly, all tumors contained varying numbers of adenoma cells with NFP-positive, dot-like areas of cytoplasmic reactivity, mostly tiny paranuclear, a finding not previously reported in human pituitary gangliocytomas. The presence of NFP in pituitary adenomas indicates neuronal differentiation in adenoma cells, suggesting a common origin for neuronal and pituitary adenoma cell elements in gangliocytomas. PMID- 16699778 TI - Tissue-specific accumulation of carotenoids in carrot roots. AB - Raman spectroscopy can be used for sensitive detection of carotenoids in living tissue and Raman mapping provides further information about their spatial distribution in the measured plant sample. In this work, the relative content and distribution of the main carrot (Daucus carota L.) root carotenoids, alpha-, beta carotene, lutein and lycopene were assessed using near-infrared Fourier transform Raman spectroscopy. The pigments were measured simultaneously in situ in root sections without any preliminary sample preparation. The Raman spectra obtained from carrots of different origin and root colour had intensive bands of carotenoids that could be assigned to beta-carotene (1,520 cm(-1)), lycopene (1,510 cm(-1)) and alpha-carotene/lutein (1,527 cm(-1)). The Raman mapping technique revealed detailed information regarding the relative content and distribution of these carotenoids. The level of beta-carotene was heterogeneous across root sections of orange, yellow, red and purple roots, and in the secondary phloem increased gradually from periderm towards the core, but declined fast in cells close to the vascular cambium. alpha-carotene/lutein were deposited in younger cells with a higher rate than beta-carotene while lycopene in red carrots accumulated throughout the whole secondary phloem at the same level. The results indicate developmental regulation of carotenoid genes in carrot root and that Raman spectroscopy can supply essential information on carotenogenesis useful for molecular investigations on gene expression and regulation. PMID- 16699780 TI - The role of positron emission tomography in pediatric musculoskeletal oncology. PMID- 16699779 TI - A diversity of serine phage integrases mediate site-specific recombination in mammalian cells. AB - This study evaluated the ability of five serine phage integrases, from phages A118, U153, Bxb1, phiFC1, and phiRV1, to mediate recombination in mammalian cells. Two types of recombination were investigated, including the ability of an integrase to mediate recombination between its own phage att sites in the context of a mammalian cell and the ability of an integrase to perform genomic integration pairing a phage att site with an endogenous mammalian sequence. We demonstrated that the A118 integrase mediated precise intra-molecular recombination of a plasmid containing its attB and attP sites at a frequency of approximately 50% in human cells. The closely related U153 integrase also performed efficient recombination in human cells on a plasmid containing the attB and attP sites of A118. The integrases from phages Bxb1, phiFC1, and phiRV1 carried out such recombination at their attB and attP sites at frequencies ranging from 11 to 75%. Furthermore, the A118 integrase mediated recombination between its attP site on a plasmid and pseudo attB sites in the human genome, i.e. native sequences with partial identity to attB. Fifteen such A118 pseudo att sites were analyzed, and a consensus recognition site was identified. The other integrases did not mediate integration at genomic sequences at a frequency above background. These site-specific integrases represent valuable new tools for manipulating eukaryotic genomes. PMID- 16699781 TI - A model of the hippocampal-cortical memory system. AB - Based on physiological evidence, we propose a theoretical model of the hippocampal-cortical memory system. The model consists of the following components: the sensory system, the hippocampus (short-term memory), and the association cortex (long-term memory). A series of key codes (local information) is supplied from the sensory system, while context (global information) is inputted from the hippocampus. The two inputs interact dynamically in the association cortex. The interactive neurons work as a detector of coincidence. The cortical network learns the memory information through the coincidence window and, finally, stores it in the form of attractors. This local-global information works as an addressor to designate the stored location of the memory in the association cortex and accelerates the process of storing and retrieving memory information. PMID- 16699782 TI - Tests of models for saccade-vergence interaction using novel stimulus conditions. AB - During natural activities, two types of eye movements - saccades and vergence - are used in concert to point the fovea of each eye at features of interest. Some electrophysiological studies support the concept of independent neurobiological substrates for saccades and vergence, namely saccadic and vergence burst neurons. Discerning the interaction of these two components is complicated by the near synchronous occurrence of saccadic and vergence components. However, by positioning the far target below the near target, it is possible to induce responses in which the peak velocity of the vertical saccadic component precedes the peak velocity of the horizontal vergence component by approximately 75 ms. When saccade-vergence responses are temporally dissociated in this way, the vergence velocity waveform changes, becoming less skewed. We excluded the possibility that such change in skewing was due to visual feedback by showing that similar behavior occurred in darkness. We then tested a saccade-related vergence burst neuron (SVBN) model proposed by Zee et al. in J Neurophysiol 68:1624-1641 (1992), in which omnipause neurons remove inhibition from both saccadic and vergence burst neurons. The technique of parameter estimation was used to calculate optimal values for responses from human subjects in which saccadic and convergence components of response were either nearly synchronized or temporally dissociated. Although the SVBN model could account for convergence waveforms when saccadic and vergence components were nearly synchronized, it could not when the components were temporally dissociated. We modified the model so that the saccadic pulse changed the parameter values of the convergence burst units if both components were synchronized. The modified model accounted for velocity waveforms of both synchronous and dissociated convergence movements. We conclude that both the saccadic pulse and omnipause neuron inhibition influence the generation of vergence movements when they are made synchronously with saccades. PMID- 16699783 TI - Simulating discrete and rhythmic multi-joint human arm movements by optimization of nonlinear performance indices. AB - An optimization approach applied to mechanical linkage models is used to simulate human arm movements. Predicted arm trajectories are the result of minimizing a nonlinear performance index that depends on kinematic or dynamic variables of the movement. A robust optimization algorithm is presented that computes trajectories which satisfy the necessary conditions with high accuracy. It is especially adapted to the analysis of discrete and rhythmic movements. The optimization problem is solved by parameterizing each generalized coordinate (e.g., joint angular displacement) in terms of Jacobi polynomials and Fourier series, depending on whether discrete or rhythmic movements are considered, combined with a multiple shooting algorithm. The parameterization of coordinates has two advantages. First, it provides an initial guess for the multiple shooting algorithm which solves the optimization problem with high accuracy. Second, it leads to a low dimensional representation of discrete and rhythmic movements in terms of expansion coefficients. The selection of a suitable feature space is an important prerequisite for comparison, recognition and classification of movements. In addition, the separate computational analysis of discrete and rhythmic movements is motivated by their distinct neurophysiological realizations in the cortex. By investigating different performance indices subject to different boundary conditions, the approach can be used to examine possible strategies that humans adopt in selecting specific arm motions for the performance of different tasks in a plane and in three-dimensional space. PMID- 16699785 TI - The facial skin lesions "see and treat" clinic: A prospective study. AB - Facial lesions are commonly referred to ear, nose and throat surgeons. Almost all are amenable to excision under local anaesthetic as a day case. However, in the UK, there is still a significant delay between referral by the general practitioner (GP) and final surgery. To address this delay, a one stop see and treat consultant led clinic was set up in the community. The aim of this study was to assess the impact of the one stop clinic on waiting times and to ascertain the satisfaction of patients with the treatment they received in this clinic. Patients with facial skin lesions were referred by the GPs to the ENT department in the usual manner. The referral letters were screened by two consultants, the appointments were booked by telephone and the patients were seen and treated in a single visit. The clinics were held in a minor surgery unit of a centrally located GP practice. Patients were seen, assessed and if the facial lesion was considered amenable to excision under local anaesthetic, the patient was consented and the procedures carried out immediately. The clinic was audited over a 1 year period. Waiting times were compared before and after the start of the project. Patients were asked to fill in a questionnaire immediately after surgery. The attendance rate was 96%. The waiting time was reduced from 121 to 47 days. Patients rated the clinic experience as excellent (88%) or good (12%) indicating a very high satisfaction rate. During the study period, 160 lesions were excised of which 22% were malignant. Patients with malignant lesions did not show any sign of recurrence at a follow up of 9 months, except in one case with basal cell carcinoma. This was operated on and removed completely. Our project shows that the aims of reducing waiting times and improving patient care were achieved with this community model of a one stop facial lesions clinic. This clinic is now an integral part of the service provided by the ear, nose and throat department at Ipswich hospital, UK. PMID- 16699786 TI - Spastic paraplegia with thin corpus callosum: description of 20 new families, refinement of the SPG11 locus, candidate gene analysis and evidence of genetic heterogeneity. AB - We studied 20 Mediterranean families (40 patients) with autosomal recessive hereditary spastic paraplegia and thin corpus callosum (ARHSP-TCC, MIM 604360) to characterize their clinical and genetic features. In six families (17 patients) of Algerian Italian, Moroccan, and Portuguese ancestry, we found data consistent with linkage to the SPG11 locus on chromosome 15q13-15, whereas, in four families (nine patients of Italian, French, and Portuguese ancestry) linkage to the SPG11 locus could firmly be excluded, reinforcing the notion that ARHSP-TCC is genetically heterogeneous. Patients from linked and unlinked families could not be distinguished on the basis of clinical features alone. In SPG11-linked kindred, haplotype reconstruction allowed significant refinement to 6 cM, of the minimal chromosomal interval, but analysis of two genes (MAP1A and SEMA6D) in this region did not identify causative mutations. Our findings suggest that ARHSP TCC is the most frequent form of ARHSP in Mediterranean countries and that it is particularly frequent in Italy. PMID- 16699788 TI - Measuring the order in ordered porous arrays: can bees outperform humans? AB - A method that explains how to quantify the amount of order in "ordered" and "highly ordered" porous arrays is derived. Ordered arrays from bee honeycomb and several from the general field of nanoscience are compared. Accurate measures of the order in porous arrays are made using the discrete radial distribution function (RDF). Nanoporous anodized aluminum oxide (AAO), hexagonal arrays from functional materials, hexagonal arrays from nanosphere lithography, and square arrays defined by interference lithography (all taken from the literature) are compared to two-dimensional model systems. These arrays have a range of pore diameters from approximately 60 to 180 nm. An order parameter, OP( 3 ) , is defined to evaluate the total order in a given array such that an ideal network has the value of 1. When we compare RDFs of man-made arrays with that of our honeycomb (pore diameter approximately 5.89 mm), a locally grown version made by Apis mellifera without the use of foundation comb, we find OP( 3 )=0.399 for the honeycomb and OP( 3 )=0.572 for man's best hexagonal array. The nearest neighbor peaks range from 4.65 for the honeycomb to 5.77 for man's best hexagonal array, while the ideal hexagonal array has an average of 5.93 nearest neighbors. Ordered arrays are now becoming quite common in nanostructured science, while bee honeycombs were studied for millennia. This paper describes the first method to quantify the order found in these arrays with a simple yet elegant procedure that provides a precise measurement of the order in one array compared to other arrays. PMID- 16699787 TI - Transcriptome analysis reveals link between proteasomal and mitochondrial pathways in Parkinson's disease. AB - There is growing evidence that dysfunction of the mitochondrial respiratory chain and failure of the cellular protein degradation machinery, specifically the ubiquitin-proteasome system, play an important role in the pathogenesis of Parkinson's disease. We now show that the corresponding pathways of these two systems are linked at the transcriptomic level in Parkinsonian substantia nigra. We examined gene expression in medial and lateral substantia nigra (SN) as well as in frontal cortex using whole genome DNA oligonucleotide microarrays. In this study, we use a hypothesis-driven approach in analysing microarray data to describe the expression of mitochondrial and ubiquitin-proteasomal system (UPS) genes in Parkinson's disease (PD). Although a number of genes showed up regulation, we found an overall decrease in expression affecting the majority of mitochondrial and UPS sequences. The down-regulated genes include genes that encode subunits of complex I and the Parkinson's-disease-linked UCHL1. The observed changes in expression were very similar for both medial and lateral SN and also affected the PD cerebral cortex. As revealed by "gene shaving" clustering analysis, there was a very significant correlation between the transcriptomic profiles of both systems including in control brains. Therefore, the mitochondria and the proteasome form a higher-order gene regulatory network that is severely perturbed in Parkinson's disease. Our quantitative results also suggest that Parkinson's disease is a disease of more than one cell class, i.e. that it goes beyond the catecholaminergic neuron and involves glia as well. PMID- 16699790 TI - Individual and multi-environment combined analyses identify QTLs for morphogenetic and reproductive development traits in white clover (Trifolium repens L.). AB - White clover (Trifolium repens L.) is a key component legume of temperate pasture agriculture and an important target for molecular marker-assisted plant breeding. A genetic map of white clover has been used to assess genetic control of agronomically important traits that vary in the F2(I.4RxI.5J) mapping family. Phenotypic analysis was performed for a range of vegetative morphogenesis traits (such as leaf area, internode length, plant height and plant spread) and reproductive morphogenesis and development traits (such as flowering date, floral intensity and seed yield), with both spatial and temporal replication. A multi environment combined analysis (combined analysis) has been performed for traits assessed across multiple experimental datasets in order to identify consistent genetic effects. Quantitative trait locus (QTLs) were detected for the majority of traits, and the locations and magnitudes of QTL effects were compared between individual and combined analyses. This molecular genetic dissection of agronomic traits in white clover provides the basis for equivalent studies in more complex populations, design of marker-assisted selection strategies and comparative genetics with model legume species. Selection for QTLs derived from the combined analysis will permit robust improvement of phenotypic traits over different environments. PMID- 16699791 TI - Assessing genetic diversity and population structure in a citrus germplasm collection utilizing simple sequence repeat markers (SSRs). AB - Twenty-four simple sequence repeat (SSR) markers were used to detect molecular polymorphisms among 370 mostly sexually derived Citrus accessions from the collection of citrus germplasm maintained at the University of California, Riverside. A total of 275 alleles were detected with an average of 11.5 alleles per locus and an average polymorphism information content of 0.625. Genetic diversity statistics were calculated for each individual SSR marker, the entire population, and for specified Citrus groups. Phylogenetic relationships among all citrus accessions and putative non-hybrid Citrus accessions were determined by constructing neighbor-joining trees. There was strong support for monophyly at the species level when hybrid taxa were removed from the data set. Both of these trees indicate that Fortunella clusters within the genus Citrus but Poncirus is a sister genus to Citrus. Additionally, Citrus accessions were probabilistically assigned to populations or multiple populations if their genotype indicated an admixture by a model-based clustering approach. This approach identified five populations in this data set. These separate analyses (distance and model based) both support the hypothesis that there are only a few naturally occurring species of Citrus and most other types of Citrus arose through various hybridization events between these naturally occurring forms. PMID- 16699792 TI - Pharmacokinetics and tumor uptake of a derivatized form of paclitaxel associated to a cholesterol-rich nanoemulsion (LDE) in patients with gynecologic cancers. AB - OBJECTIVE: A cholesterol-rich nanoemulsion termed LDE concentrates in cancer tissues after injection into the bloodstream. The association of a derivatized paclitaxel to LDE showed lower toxicity and increased antitumoral activity as tested in a B16 melanoma murine model. Here, the pharmacokinetics of LDE paclitaxel oleate and the ability of LDE to concentrate the drug in the tumor were investigated in patients with gynecologic cancers. METHODS: Either LDE paclitaxel oleate doubly labeled with [(14)C]-cholesteryl oleate and [(3)H] paclitaxel oleate or [(3)H]-paclitaxel-cremophor was intravenously injected into eight patients. Blood samples were collected over 24 h to determine the plasma decay curves. Fractional clearance rate (FCR) and pharmacokinetic parameters were calculated by compartmental analysis. Also, specimens of tumors and the corresponding normal tissues were excised during the surgery for radioactivity measurement. RESULTS: The LDE and paclitaxel oleate FCR were similar (0.092 +/- 0.039 and 0.069 +/- 0.027 h(-1), respectively, n = 5, P = 0.390). FCR of paclitaxel oleate associated to LDE was smaller than that of paclitaxel-cremophor (0.231 +/- 0.128 h(-1), P = 0.028). Paclitaxel oleate T (1/2 )and AUC were greater than those of paclitaxel-cremophor (T (1/2 )=( )14.51 +/- 3.23 and 6.62 +/- 2.05 h and AUC = 2.49 +/- 0.35 and 1.26 +/- 0.40, respectively, P = 0.009, P = 0.004). The amount of paclitaxel and LDE-radioactive labels in the tumor was 3.5 times greater than in the normal tissues. CONCLUSION: Paclitaxel oleate associated to LDE is stable in the bloodstream and has greater plasma half-life and AUC than those for paclitaxel-cremophor. LDE concentrates 3.5 times more paclitaxel in malignant tissues than in normal tissues. Therefore, association to LDE is an interesting strategy for using paclitaxel to treat gynecologic cancers. PMID- 16699793 TI - Combination therapy of human pancreatic cancer implanted in nude mice by oral fluoropyrimidine anticancer agent (S-1) with interferon-alpha. AB - PURPOSE: We evaluated the antitumor and antiangiogenic activities of human natural interferon-alpha (IFN-alpha) alone or in combination with S-1 against human pancreatic cancer cells. METHODS: Three days after the subcutaneous (s.c.) implantation of tumor cells, mice (n = 12) were received s.c. injection with IFN alpha alone (10,000 U six times a week), oral administration with S-1 alone (8 mg/kg six times a week), or both with IFN-alpha and S-1 (8, 10, 12 mg/kg six times a week). RESULTS: Administration of IFN-alpha in combination with S-1 significantly decreased progressive growth and angiogenesis of human pancreatic cancer cells. The combination therapy produced more significant inhibition in expression of the representative proangiogenic molecules, vascular endothelial growth factor and basic fibroblast growth factor than individual treatment either IFN-alpha or S-1 alone did. These treatments also decreased the staining of proliferating cell nuclear antigen, induced apoptosis and decreased microvessel density. In order to better understand the precise molecular mechanisms by which IFN-alpha and S-1 exert its effects, we have utilized cDNA microarray including 124 known genes to determine the gene expression profile altered by IFN-alpha and S-1 treatment. We found a total of seven genes which showed a twofold change after IFN-alpha and S-1 treatment in addition to VEGF, bFGF, CD31, MMP-2, MMP-7 and MMP-9. Among these genes, we found down-regulation of six genes and up regulation of one gene, which are related to angiogenesis, tumor cell invasion and metastasis. CONCLUSIONS: These data suggest that administration of IFN-alpha in combination with S-1 may provide a novel and effective approach to the treatment of human pancreatic cancer. PMID- 16699794 TI - Extraocular sebaceous carcinoma in Muirr Torre Syndrome with unfavorable prognosis. PMID- 16699795 TI - The evolution of photoselective vaporization prostatectomy (PVP): advancing the surgical treatment of benign prostatic hyperplasia. PMID- 16699796 TI - Rupture of rudimentary uterine horn pregnancy at 37 weeks gestation with fetal survival. AB - Pregnancy in rudimentary uterine horn is rare and associated with maternal and fetal mortality and morbidity. This patient presented with unexplained abdominal pain antenataly. Laparotomy for suspected abruption at 37 weeks revealed ruptured rudimentary horn with placenta accreta. A subtotal hemihysterectomy was performed. The result was fetal salvage in good condition. PMID- 16699799 TI - Transfer of reboxetine into breastmilk, its plasma concentrations and lack of adverse effects in the breastfed infant. AB - OBJECTIVE: To investigate the transfer of reboxetine into milk, the absolute and relative infant doses via milk and to assess plasma concentrations and adverse unwanted effects in the breastfed infant. METHODS: Multiple samples of blood and milk were obtained over a dose interval at steady-state from four women who were taking reboxetine for postnatal depression. Drug concentrations in plasma and milk were measured by high performance liquid chromatography and milk/plasma ratio (M/P), absolute infant dose and relative infant dose were estimated by standard methods. Their four, breastfed, infants were also examined clinically, and a blood sample was taken for drug analysis. RESULTS: The median (range) dose taken by the women was 6 (4-10) mg/day. There was no significant difference in reboxetine concentration between paired fore-and hind-milk samples. The mean (95% CI) M/P was 0.06 (0.03, 0.09). Absolute infant dose was 1.7 (0.7, 2.4) microg/kg/day for reboxetine while the relative infant dose was 2.0% (1.3, 2.7%). Three of the infants met normal developmental milestones and no adverse effects were seen in any infant. The fourth infant had developmental problems that were not associated with the maternal reboxetine therapy. The concentrations of reboxetine in plasma from the four infants were <4 microg/l, 2.6 microg/l, 2.3 microg/l and 5 microg/l, respectively. CONCLUSION: The study suggests that reboxetine use by lactating women is safe for the breastfed infant. Nevertheless, our study had only four mother/baby pairs, and each decision to breastfeed should always be made on the basis of an individual risk/benefit analysis. PMID- 16699800 TI - Primary care physicians' behaviors towards risk of iatrogenesis in elderly patients. AB - OBJECTIVES: Iatrogenesis is common in elderly patients. This phenomenon could be reduced by improving awareness of general practitioners (GPs). We studied GPs' prescribing behavior to elderly patients, to identify GP and patient characteristics related to cautious prescribing behaviors. METHODS: The observational study sampled 106 GPs who had consecutively recruited, on average, 12 patients over 75 years of age. GPs completed a questionnaire on their practice and a questionnaire for each patient they recruited, describing their usual care of elderly patients, and criteria for prescribing therapy. GPs' behaviors were studied with five scores and a global prescribing behavior (adequate/inadequate). The relationship between patients' risk of iatrogenesis and GPs' behaviors was studied with hierarchical logistic models. RESULTS: A total of 106 GPs recruited 1,318 patients (mean age 80.7 years, 36% males). A wide variety of behaviors was observed among GPs. Only 40% of GPs had specific prevention-oriented visits, while only 19.2% considered that prevention was optimally implemented in their practice. On average, GPs had behaviors considered adequate in about half of encounters. GPs' global behaviors were more likely to be adequate for patients at higher risk (OR=1.47, 95%CI: 1.10-1.95). Likewise, before prescribing to patients at higher risk of iatrogenesis, GPs were more likely to collect data on financial autonomy, on clinical/biological data, and to adopt good prescription practices, while less attention was paid to patients' physical and psychological autonomy. CONCLUSIONS: GPs tended to be more cautious when prescribing to patients at higher iatrogenesis risk. However, overall prescribing behaviors were not optimal. Efforts are needed to improve the quality of care in elderly patients. PMID- 16699801 TI - Oncolytic murine autonomous parvovirus, a candidate vector for glioma gene therapy, is innocuous to normal and immunocompetent mouse glial cells. AB - The sensitivity of brain tumour cells to wild-type or recombinant parvoviruses H1 PV and MVMp makes these agents promising candidates for gene therapy of astrocytoma. This application raises the question of whether parvoviruses exert deleterious or bystander effects on normal glial cells surrounding tumours. We addressed this question in the mouse model by using cell cultures derived from BALB/c, C57BL/6 and VM/Dk strains. Astrocytes and a large proportion of microglia cultures were competent for MVMp uptake. Infection was, however, abortive as replication-associated viral proteins synthesis took place in less than 10% of astrocytes and no progeny virions were produced. This restriction was even more pronounced for microglia in which no viral protein expression could be detected, save for a minute fraction of VM/Dk-derived cells. Infection with MVMp had no significant effect on glial cell survival and did not interfere with their immune potential. Indeed, neither the lipopolysaccharide (LPS)/interferon (IFN-gamma) induced cytotoxicity of VM/Dk-derived microglia towards the mouse glioma (MT539MG) cell line, nor the glial cells capacity for tumour necrosis factor alpha production upon LPS stimulation or LPS/IFN-gamma stimulation were affected by infection with MVMp. Moreover, stimulation with LPS and/or IFN-gamma resulted in a decreased expression of the viral replicative and cytotoxic protein NS1. Together, our data indicate that, in the natural host, a majority of normal glial cells are not competent for MVMp replication and that the abortive infection taking place in a minor fraction of these cells fails to impede their survival and immunocompetence, giving credit to the consideration of autonomous parvoviruses for glioma therapy. PMID- 16699802 TI - The effect of MR contrast medium dose on pituitary gland enhancement, microlesion enhancement and pituitary gland-to-lesion contrast conspicuity. AB - INTRODUCTION: The purpose of this study was to compare the differences in gland enhancement, microlesion enhancement and gland-lesion contrast ratio in patient groups in which half-dose (HD), standard-dose (SD) and double-dose (DD) contrast medium was used in pituitary MR imaging. METHODS: Pituitary gland enhancement and microlesion enhancement were measured and gland-lesion contrast ratios were calculated in 18 patients receiving HD (0.05 mmol/kg), 9 receiving SD (0.1 mmol/kg) and 13 receiving DD (0.2 mmol/kg) contrast medium. Gland enhancement and microlesion enhancement over baseline were determined employing DICOM region of interest measurements and compared after normalization to temporal lobe white matter. Contrast ratios and differences were also calculated and compared. RESULTS: Gland enhancement and lesion enhancement were greater with larger contrast medium doses (gland: HD 50%, SD 99%, DD 132%; microlesion: HD 19%, SD 54%, DD 86%). The gland-lesion contrast ratios were similar with the three doses (25.6%), reflecting expected similar fractional contrast medium distributions in spite of different doses. The signal difference between gland and microlesion, therefore, was a fixed percentage of gland enhancement (DeltaS approximately 26%) with greater signal differences with larger contrast medium doses. CONCLUSION: Greater gland-to-lesion signal differences with larger contrast medium doses would likely improve pituitary microlesion visualization and margin characterization aiding in microlesion detection as well as preoperative planning. PMID- 16699803 TI - Prehospital triage and survival of major trauma patients in a Dutch regional trauma system: relevance of trauma registry. AB - BACKGROUND AND AIMS: Since 1999, the Dutch trauma care has been regionalized into ten trauma systems. This study is the first to review such a trauma system. The aim was to examine the sensitivity of prehospital triage criteria [triage revised trauma score (T-RTS)] in identifying major trauma patients and to evaluate the current level of trauma care of a regionalized Dutch trauma system for major trauma patients. PATIENTS AND METHODS: Major trauma patients (n=511) (June 2001 December 2003) were selected from a regional trauma registry database. The prehospital T-RTS was computed and standardized W scores (Ws) were generated to compare observed vs expected survival based on contemporary US- and UK-norm databases. RESULTS: The T-RTS showed low sensitivity for the prehospital identification of major trauma patients [34.1% (T-RTS< or =10)]. Nevertheless, 78.0% of all major trauma patients were directly managed by the trauma center. These patients were more severely injured than their counterparts at non-trauma center hospitals (p<0.001). No significant difference emerged between the mortality rates of both groups. The Ws {-0.46 calculated on the US model [95% confidence interval (CI) ranging from -1.99 to 1.07]} [0.60 calculated on the UK model (95% CI ranging from -1.25 to 2.44)] did not differ significantly from zero. CONCLUSION: The trauma center managed most of the major trauma patients in the trauma system but the triage criteria need to be reconsidered. The level of care of the regional trauma system was shown to measure up to US and UK benchmarks. PMID- 16699804 TI - Cerebrospinal fluid obstruction and malabsorption in human neonatal hydrocephaly. AB - INTRODUCTION: The pathophysiology involved in human neonatal high-pressure hydrocephalus (HC) includes both cerebrospinal fluid (CSF) malabsorption and obstruction. OBJECTIVE: The aim was to estimate the relative contribution between CSF malabsorption and obstruction in three different etiological groups of neonatal high-pressure HC by assessment of specific CSF biomarkers indicative of growth factor- and fibrosis-related CSF malabsorption (transforming growth factor beta-1 (TGF beta-1), aminoterminal propeptide of type 1 collagen (PC1NP)]. MATERIALS AND METHODS: Patients were subdivided into three groups. Group A: spina bifida HC (n=12); group B: non-haemorrhagic triventricular HC (n=4); and group C: posthaemorrhagic HC (n=6). To exclude for confounding differences in pro inflammatory state between the three groups, interleukin-6 (IL-6) CSF concentrations were assessed. Consecutively, the CSF concentrations of TGF beta-1 and PC1NP were compared between the different groups. RESULTS: Median CSF concentrations of IL-6 were low and did not differ between groups. Median CSF concentrations of PC1NP were significantly lower in group A (median: 180 ng/ml, range 90-808) than in group C (median: 1,060, range 396-1194; p=0.002). TGF beta 1 concentrations were significantly higher in group C (median 355 pg/ml, range 129-843) than in groups A (median 103, range 78-675 pg/ml) and B (median 120 pg/ml, range 91-188; p=0.01 and 0.03, respectively). CONCLUSIONS: In neonatal posthaemorrhagic HC, high concentrations of malabsorption-related biomarkers contrast with lower concentrations in SB and non-haemorrhagic triventricular HC. During the early development of high pressure HC in SB neonates, CSF biomarkers strongly indicate that CSF obstruction contributes more to the development of HC than malabsorption. PMID- 16699805 TI - Monitoring motor function during resection of tumours in the lower brain stem and fourth ventricle. AB - OBJECTIVES: Even in the days of modern microsurgery, the removal of a brain stem lesion remains a surgical challenge. Especially when operating on children, the prognosis is directly related to the radicality of the resection; however, a radical resection is often associated with surgical morbidity. Intraoperative neuromonitoring could help to minimise the surgical morbidity, but few studies have been performed to clarify the value of this monitoring. We investigated a prospective series of 21 patients with lesions involving the brain stem for the prognostic value and benefits of neuromonitoring. METHODS: We performed intraoperative neuromonitoring of cranial nerve function by electromyography (EMG) and motor evoked potential (MEP). The results were correlated with postoperative neurological deficits. CONCLUSIONS: There is a good correlation between intraoperative neurophysiological events and postoperative neurological deficits in patients with lesions of the brain stem. In general, transient, prolonged, spontaneous activity in EMG is associated with a transient paresis of the respective muscle, whereas a permanent spontaneous activity is associated with a permanent deficit. Intraoperative neuromonitoring reliably predicts postoperative neurological function in patients with tumours of the lower brain stem and fourth ventricle. This neuromonitoring guides the neurosurgeon in the operation and may decrease surgical morbidity. We recommend using monitoring of MEP and EMG of the lower cranial nerves in surgery on all patients with lesions involving the lower brain stem and fourth ventricle. PMID- 16699806 TI - Osteogenesis induced by autologous bone marrow cells transplant in the pediatric skull. AB - BACKGROUND AND PURPOSE: The ability of cranial bone to repair defects of continuity is limited and it is mostly dependent on the age of the patient. In infancy and in early pediatric age, the scarce thickness of the calvarial bones and the need for a harmonic development of the child's skull limit the application of most of the surgical procedures usually utilized in older patients. We tested the ability of mononucleated cells, derived from the patient's bone marrow and transplanted on the site of the cranial bone defect, to increase the rate of mineralization of the autologous osteogenesis to obtain the complete restoration of the skull continuity. METHOD: Four children, aged 26, 28, 37, and 79 months, respectively, affected by a stabilized and persistent cranial bone defect of posttraumatic or postsurgical origin, were treated. A sandwich shaped shell, made of extrused absorbable polylactic copolymers material, was used to hold in place a freeze-dried mineralized collagen matrix associated with a nonceramic hydroxyapatite scaffold, where autologous bone marrow mononucleated cells were inseminated. RESULTS: In all patients, a rapid autologous bone osteogenesis was observed with a clear dimensional reduction of the bone defect few months after the autologous bone marrow cells seeding. CONCLUSIONS: The preliminary results of this research suggest the use of autologous bone marrow cells to increase the autologous osteogenesis in early pediatric age in cases in which correction of skull bone defects is best realized with autologous bone. PMID- 16699808 TI - Antimalarial drugs: recent advances in molecular determinants of resistance and their clinical significance. AB - Molecular determinants of antimalarial drug resistance are useful and informative tools that complement phenotypic assays for drug resistance. They also guide the design of strategies to circumvent such resistance once it has reached levels of clinical significance. Established resistance to arylaminoalcohols such as mefloquine and lumefantrine in SE Asia is mediated primarily by gene amplification of the P. falciparum drug transporter, pfmdr1. Single nucleotide polymorphisms in pfmdr1, whether assessed in field isolates or transfection experiments, are associated with changes in IC(50) values (to arylaminoalcohols and chloroquine), but not of such magnitude as to influence clinical treatment outcomes. Recently described emerging in vitro resistance to artemisinins in certain areas correlates with mutations in the SERCA-like sequence PfATP6 and supports PfATP6 as a key target for artemisinins. PMID- 16699807 TI - Prediction of obstructive sleep apnea in patients presenting to a tertiary care center. AB - The objective of this prospective observational clinical study is to derive and validate a diagnostic model for prediction of obstructive sleep apnea (OSA) in subjects presenting with non-sleep-related complaints in a tertiary care center in north India. We included 102 subjects (group I, range 31-70 years) presenting to the hospital with non-sleep-related complaints. None of the subjects had any significant comorbid illness such as respiratory or congestive cardiac failure. All subjects underwent detailed evaluation including polysomnography (PSG). Various parameters were compared between the cases (apnea-hypopnea index, AHI > or =15/h) and controls (AHI <15/h). Using multivariate logistic regression analysis, a diagnostic model for prediction of OSA was derived. Subsequently, using similar selection criteria, 104 subjects (group II, range 32-68 years) were included for validation of the newly derived diagnostic model. Body mass index [BMI; OR (95% CI), 1.14(1.1-1.2)], male gender 5.0(1.4-27.1), relative-reported snoring index (SI) 2.8(1.7-5.0), and choking index (ChI) 8.1(1.4-46.5) were significant, independent predictors of OSA. Diagnostic model was computed as score = [1.61 x (gender)] + [1.01 x (S1)] + [2.09 x (ChI)] + [0.1 x (BMI)] where, gender: 0 = female, 1 = male and SI, ChI, BMI are actual values. The diagnostic model had an area under the receiver operator characteristics curve of 89.6%. A cutoff of 4.3 for the score was associated with sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of 91.3, 68.5, 70.5, and 92.3%, respectively. Misclassification rate with the application of the diagnostic model on group II subjects was 13.5% (14/104). Sensitivity, specificity, PPV, and NPV of the model for predicting OSA in this group were 82, 90.7, 89.1, and 84.5%, respectively. BMI, male gender, SI, and ChI are independent predictors of OSA. Diagnostic model derived from these parameters is useful for predicting presence of OSA and screening subjects for PSG. PMID- 16699810 TI - Skeletal muscle progenitor cells: from embryo to adult. PMID- 16699809 TI - The roles of cannabinoid and dopamine receptor systems in neural emotional learning circuits: implications for schizophrenia and addiction. AB - Cannabinoids represent one of the most widely used hallucinogenic drugs and induce profound alterations in sensory perception and emotional processing. Similarly, the dopamine (DA) neurotransmitter system is critical for the central processing of emotion and motivation. Functional disturbances in either of these neurotransmitter systems are well-established correlates of the psychopathological symptoms and behavioral manifestations observed in addiction and schizophrenia. Increasing evidence from the anatomical, pharmacological and behavioral neuroscience fields points to complex functional interactions between these receptor systems at the anatomical, pharmacological and neural systems levels. An important question relates to whether these systems act in an orchestrated manner to produce the emotional processing and sensory perception deficits underlying addiction and schizophrenia. This review describes evidence for functional neural interactions between cannabinoid and DA receptor systems and how disturbances in this neural circuitry may underlie the aberrant emotional learning and processing observed in disorders such as addiction and schizophrenia. PMID- 16699811 TI - Neurotrophin signaling: many exciting surprises! AB - Neurotrophins are growth factors implicated in the development and maintenance of different neuronal populations in the nervous system. Neurotrophins bind to two sets of receptors, Trk receptor tyrosine kinases and the p75NTR receptor, to activate several different signaling pathways that mediate various biological functions. While Trk receptor activation has been well-studied and triggers the well-characterized Ras/Rap-MAPK, PI3K-Akt, and PLCgamma-PKC cascades, p75NTR signaling is more complex, and its in vivo significance has not yet been completely determined. In the last few years, p75NTR has received much attention mainly due to recent findings describing pro-neurotrophins as new ligands for the receptor and the ability of the receptor to form different complexes with other transmembrane proteins. This review will update the neurotrophin signaling pathways known for Trk receptors to include newly identified Trk-interacting molecules and will address surprising new findings that suggest a role for p75NTR in different receptor complexes and functions. PMID- 16699812 TI - The cellular functions of clathrin. AB - Membranes and proteins are moved around the cell in small vesicles. A protein coat aids the budding of such vesicles from donor membranes. The major type of coat used by the cell is composed of clathrin, a three-legged protein that can form lattice-like coats on membranes destined for trafficking. In this review, I outline what we know about clathrin and discuss some recent advances in understanding the basic biology of this fascinating molecule, which include building a molecular model of a clathrin lattice and discovery of a new function for clathrin that occurs during mitosis. PMID- 16699813 TI - Light-driven DNA repair by photolyases. AB - DNA photolyases are highly efficient light-driven DNA repair enzymes which revert the genome-damaging effects caused by ultraviolet (UV) radiation. These enzymes occur in almost all living organisms exposed to sunlight, the only exception being placental mammals like humans and mice. Their catalytic mechanism employs the light-driven injection of an electron onto the DNA lesion to trigger the cleavage of cyclobutane- pyrimidine dimers or 6-4 photoproducts inside duplex DNA. Spectroscopic and structural analysis has recently yielded a concise view of how photolyases recognize these DNA lesions involving two neighboring bases, catalyze the repair reaction within a nanosecond and still achieve quantum efficiencies of close to one. Apart from these mechanistic aspects, the potential of DNA photolyases for the generation of highly UV-resistant organisms, or for skin cancer prevention by ectopical application is increasingly recognized. PMID- 16699814 TI - Treatment of ADHD with French maritime pine bark extract, Pycnogenol. AB - Attention Deficit/Hyperactivity Disorder (ADHD) is the most common psychiatric disorder in children. Pycnogenol, an extract from the bark of the French maritime pine, consisting of phenolic acids, catechin, taxifolin and procyanidins, has shown improvement of ADHD in case reports and in an open study. Aim of the present study was to evaluate the effect of Pycnogenol on ADHD symptoms. Sixty one children were supplemented with 1 mg/kg/day Pycnogenol or placebo over a period of 4 weeks in a randomised, placebo-controlled, doubleblind study. Patients were examined at start of trial, 1 month after treatment and 1 month after end of treatment period by standard questionnaires: CAP (Child Attention Problems) teacher rating scale, Conner's Teacher Rating Scale (CTRS), the Conner's Parent Rating Scale (CPRS) and a modified Wechsler Intelligence Scale for children. Results show that 1-month Pycnogenol administration caused a significant reduction of hyperactivity, improves attention and visual-motoric coordination and concentration of children with ADHD. In the placebo group no positive effects were found. One month after termination of Pycnogenol administration a relapse of symptoms was noted. Our results point to an option to use Pycnogenol as a natural supplement to relieve ADHD symptoms of children. PMID- 16699815 TI - The effect of gender in diagnosing early schizophrenia--an experimental case simulation study. AB - BACKGROUND: Studies of diagnostic practice confirm that there is a diagnostic delay in diagnosing women with schizophrenia compared to diagnosing men. The aim of the present study was to investigate the diagnostic practice of Norwegian and Russian psychiatrists when it comes to early psychosis, emphasising gender differences. We wanted to study the association between patient gender as such and diagnostic decision-making among psychiatrists. MATERIALS AND METHODS: Psychiatrists in Norway and the Archangels region in Russia were invited to participate in a study of diagnostic practice, and received a written case description of a patient with early psychosis symptoms that could be interpreted as schizophrenia. They were, however, not informed that 50% of them received a female case description and 50% a male case description. Apart from the patient being described as "he" or "she" the stories were identical. Effects of patient gender, clinician gender, age and main area of interest were estimated using logistic regression analysis. RESULTS: A total of 467 psychiatrists answered the questionnaire. We found that schizophrenia diagnosis was given significantly more often to the male case than to the female case. Our finding remained significant after adjustment for country, clinician gender, age and main area of interest, and is unlikely to be explained by known biases. CONCLUSION: Patient gender in itself affects clinicians' diagnostic practice regarding schizophrenia, as schizophrenia diagnosis is given significantly more often to a male case description than to a female one, the descriptions being otherwise identical. PMID- 16699816 TI - The role of group I metabotropic glutamate receptors in schizophrenia. AB - It has been proposed that glutamatergic transmission, in particular NMDA receptor function, might be altered in schizophrenia. This hypothesis is mainly based on the observation that uncompetitive NMDA receptor antagonists, e.g. phencyclidine, evoke psychotic symptoms in healthy subjects, whereas agonists interacting at the glycine site of the NMDA receptor complex, e.g. glycine or D-serine, administered jointly with typical neuroleptics, can alleviate schizophrenic symptoms. The function of NMDA receptors may be modulated by group I mGluRs (mGluR1 and mGluR5), which have also been shown to be altered in schizophrenia. In rodents, mGluR5 antagonists, but not mGluR1 ones, potentiate the locomotor activity and the deficit of prepulse inhibition (PPI) induced by uncompetitive NMDA receptor antagonists. These antagonists (of either type) administered alone are not active in the above tests. Hence, antagonists of mGluR1 and mGluR5 may evoke different effects on the NMDA receptor antagonists-induced behavior and, possibly, on schizophrenic symptoms. PMID- 16699817 TI - An influence of ligands of metabotropic glutamate receptor subtypes on parkinsonian-like symptoms and the striatopallidal pathway in rats. AB - Several data indicate that inhibition of glutamatergic transmission may be important to alleviate of parkinsonian symptoms. Therefore, the aim of the present paper is to review recent studies on the search for putative antiparkinsonian-like effects of mGluR ligands and their brain targets. In order to inhibit glutamatergic transmission, the group I mGluRs (mGluR1 and mGluR5) were blocked, and group II (mGluR2/3) or III (mGluR4/7/8) mGluRs were activated. Systemic or intrastriatal administration of group I mGluR antagonists (mGluR5 - MPEP, MTEP; mGluR1 - AIDA) was found to inhibit parkinsonian-like symptoms (catalepsy, muscle rigidity) in rats. MPEP administered systemically and mGluR1 antagonists (AIDA, CPCCOEt, LY367385) injected intrastriatally reversed also the haloperidol-increased proenkephalin (PENK) mRNA expression in the striatopallidal pathway. Similarly, ACPT-1, a group III mGluR agonist, administered into the striatum, globus pallidus or substantia nigra inhibited the catalepsy. Intrastriatal injection of this compound reduced the striatal PENK expression induced by haloperidol. In contrast, a group II mGluR agonist (2R,4R-APDC) administered intrastriatally reduced neither PENK expression nor the above mentioned parkinsonian-like symptoms. Moreover, a mixed mGluR8 agonist/AMPA antagonist, (R,S)-3,4-DCPG, administered systemically evoked catalepsy and enhanced both the catalepsy and PENK expression induced by haloperidol. The results reviewed in this article seem to indicate that group I mGluR antagonists or some agonists of group III may possess antiparkinsonian properties, and point at the striatopallidal pathway as a potential target of therapeutic intervention. PMID- 16699818 TI - Chlorpromazine, clozapine and olanzapine inhibit anionic amino acid transport in cultured human fibroblasts. AB - We report here that chlorpromazine, a first generation antipsychotic drug, inhibits anionic amino acid transport mediated by system X(-) (AG) (EAAT transporters) in cultured human fibroblasts. With 30 microM chlorpromazine, transport inhibition is detectable after 3 h of treatment, maximal after 48 h (>60%), and referable to a decrease in V(max). Chlorpromazine effect is not dependent upon changes of membrane potential and is selective for system X(-) (AG) since transport systems A and y(+) are not affected. Among antipsychotic drugs, the inhibitory effect of chlorpromazine is shared by two dibenzodiazepines, clozapine and olanzapine, while other compounds, such as risperidon, zuclopentixol, sertindol and haloperidol, are not effective. Transport inhibition by clozapine and olanzapine, but not by chlorpromazine, is reversible, suggesting that the mechanisms involved are distinct. These results indicate that a subset of antipsychotic drugs inhibits EAAT transporters in non nervous tissues and prompt further investigation on possible alterations of glutamate transport in peripheral tissues of schizophrenic patients. PMID- 16699819 TI - The role of L-arginine in toxic liver failure: interrelation of arginase, polyamine catabolic enzymes and nitric oxide synthase. AB - The existing interrelation in metabolic pathways of L-arginine to polyamines, nitric oxide (NO) and urea synthesis could be affected in sepsis, inflammation, intoxication and other conditions. The role of polyamines and NO in the toxic effect of mercury chloride on rat liver function was studied. Administration of mercury chloride for 24 h led to significantly elevated plasma activities of Alanine transaminase (ALT) and Aspartate transaminase (AST). Malondyaldehyde (MDA) levels were unaffected (p > 0.05) and arginase activity was significantly decreased (p < 0.05) while nitrate/nitrite production was significantly elevated (p < 0.001) in liver tissue. Polyamine oxidase (PAO) and diamine oxidase (DAO) activities, enzymes involved in catabolism of polyamines, were decreased. L arginine supplementation to intoxicated rats potentiated the effect of mercury chloride on NO production and it was ineffective on arginase activity. Results obtained in this study show that mercury chloride-induced toxicity leads to abnormally high levels of ALT and AST that may indicate liver damage with the involvement of polyamine catabolic enzymes and NO. PMID- 16699822 TI - Age-related changes of alpha-crystallin aggregate in human lens. AB - Lens alpha-crystallin, composed of two subunits alpha A- and alpha B-crystallin, forms large aggregates in the lens of the eye. The present study investigated the aggregate of human lens alpha-crystallin from elderly and young donors. Recombinant alpha A- and alpha B-crystallins in molar ratios of alpha A to alpha B at 1:1, corresponding to the aged sample, were also studied in detail. We found by ultra-centrifugation analysis that the alpha-crystallin aggregate from elderly donors was large and heterogeneous with an average sedimentation coefficient of 30 S and a range of 20-60 S at 37 degrees C. This was higher compared to the young samples that had an average sedimentation coefficient of 17 S. The sedimentation coefficients of recombinant alpha A- and alpha B-crystallins were approximately 12 S and 15 S, respectively. Even when recombinant alpha crystallins were mixed in molar ratios equivalent to those found in vivo, similar S values as the native aged alpha-crystallin aggregates were not obtained. Changes in the self-association of alpha-crystallin aggregate were correlated to changes in chaperone activity. Alpha-crystallin from young donors, and recombinant alpha A- and alpha B-crystallin and their mixtures showed chaperone activity, which was markedly lost in samples from the aged alpha-crystallin aggregates. PMID- 16699821 TI - Enhancement of transglutaminase activity and polyamine depletion in B16-F10 melanoma cells by flavonoids naringenin and hesperitin correlate to reduction of the in vivo metastatic potential. AB - The in vitro and in vivo effects of two flavonons, naringenin (NG) and hesperitin (HP) on the proliferation rate of highly metastatic murine B16-F10 melanoma cell were investigated. NG or HP treatment of melanoma cells produced a remarkable reduction of cell proliferation, paralleled with both the lowering of the intracellular levels of polyamine, spermidine and spermine and the enhancement of transglutaminase (TGase, EC 2.3.2.13) activity. Orally administered NG or HP in C57BL6/N mice inoculated with B16-F10 cells affected the pulmonary invasion of melanoma cells in an in vivo metastatic assay. The number of lung metastases detected by a computerized image analyzer was reduced, compared to untreated animals, by about 69% in NG-treated mice and by about 36% in HP-treated mice. Survival studies showed that 50% of the NG-treated animals died 38 +/- 3.1 days after tumor cell injection (control group: 18 +/- 1.5 days) and HP-treated mice died 27 +/- 2.3 days after cell inoculation. Taken together, these findings provide further evidences for the potential anticancer properties of dietary flavonoids as chemopreventive agents against malignant melanoma. PMID- 16699824 TI - Substrate specificity of the amino acid transporter PAT1. AB - The proton coupled amino acid transporter PAT1 expressed in intestine, brain, and other organs accepts L- and D-proline, glycine, and L-alanine but also pharmaceutically active amino acid derivatives such as 3-amino-1-propanesulfonic acid, L-azetidine-2-carboxylic acid, and cis-4-hydroxy-D-proline as substrates. We systematically analyzed the structural requirements for PAT1 substrates by testing 87 amino acids, proline homologs, indoles, and derivatives. Affinity data and effects on membrane potential were determined using Caco-2 cells. For aliphatic amino acids, a blocked carboxyl group, the distance between amino and carboxyl group, and the position of the hydroxyl group are affinity limiting factors. Methylation of the amino group enhances substrate affinity. Hetero atoms in the proline template are well tolerated. Aromatic alpha-amino acids display low affinity. PAT1 interacts strongly with heterocyclic aromatic acids containing an indole scaffold. The structural requirements of PAT1 substrates elucidated in this study will be useful for the development of prodrugs. PMID- 16699825 TI - y+ LAT-1 mediates transport of the potent and selective iNOS inhibitor, GW274150, in control J774 macrophages. AB - This study has characterised the transport mechanism(s) for the novel and selective inhibitor of inducible nitric oxide synthase (iNOS), GW274150, in murine macrophage J774 cells. Transport of GW274150 was saturable (K(m) = 0.24 +/ 0.01 mM and V(max) of 8.5 +/- 0.12 pmol.microg protein(-1) min(-1)), pH insensitive and largely Na(+)-independent. Transport was also susceptible to trans-stimulation and was significantly inhibited by a 10-fold excess of L arginine, L-lysine, L-leucine, L-methionine, L-glutamine and 6-diazo-5-oxo-L norleucine but not by other amino acids or by N-ethylmaleimide. More importantly, the inhibitions caused by the neutral amino acids were critically dependent on Na(+). These results strongly implicate system y(+)L in the transport of GW274150. Northern blot analysis confirmed this by revealing the presence of transcripts for y(+)LAT-1 but not y(+)LAT-2. Thus, taken together, our data show for the first time that J774 macrophages express y(+)LAT-1 transporters and that these carriers mediate transport of GW2741500 at least in these cells. PMID- 16699826 TI - Long-term tryptophan administration enhances cognitive performance and increases 5HT metabolism in the hippocampus of female rats. AB - It has been shown in various studies that increase in serotonergic neurotransmission is associated with increased memory consolidation whereas low brain 5HT impairs memory performance. In the first phase of our study we found that tryptophan (TRP) administration for 6 weeks increased plasma TRP and whole brain TRP, 5HT and 5HIAA levels. Many brain regions are involved in the learning process but particularly the hippocampus is known to have key role in learning and memory. The present study was therefore designed to investigate the effects of TRP loading particularly on hippocampal 5HT metabolism and cognitive performance in rats. TRP-treated rats demonstrated spatial enhancement as evidenced by a significant decrease in time to find the hidden food reward in radial arm maze test (RAM). The important finding of the present study was the greater increase in the 5HT metabolism in hippocampus than in any other brain region of the TRP-treated rats. This increased 5HT metabolism in the hippocampus emphasizes the involvement of this region in memory process. PMID- 16699827 TI - A combination of caffeine and taurine has no effect on short term memory but induces changes in heart rate and mean arterial blood pressure. AB - Red Bull energy drink has become extraordinarily popular amongst college students for use as a study aid. We investigated the combined effects of Red Bull's two active ingredients, caffeine and taurine, on short term memory. Studies on the effects of these two neuromodulators on memory have yielded mixed results, and their combined actions have not yet been investigated. In this double-blind study, college student subjects consumed either caffeine and taurine pills or a placebo and then completed a memory assessment. Heart rate and blood pressure were monitored throughout the testing period. The combination of caffeine and taurine had no effect on short term memory, but did cause a significant decline in heart rate and an increase in mean arterial blood pressure. The heart rate decline may have been caused by pressure-induced bradycardia that was triggered by caffeine ingestion and perhaps enhanced by the actions of taurine. PMID- 16699828 TI - Acetohydroxyacid synthase and its role in the biosynthetic pathway for branched chain amino acids. AB - The branched-chain amino acids are synthesized by plants, fungi and microorganisms, but not by animals. Therefore, the enzymes of this pathway are potential target sites for the development of antifungal agents, antimicrobials and herbicides. Most research has focused upon the first enzyme in this biosynthetic pathway, acetohydroxyacid synthase (AHAS) largely because it is the target site for many commercial herbicides. In this review we provide a brief overview of the important properties of each enzyme within the pathway and a detailed summary of the most recent AHAS research, against the perspective of work that has been carried out over the past 50 years. PMID- 16699829 TI - The complete genome sequence of pepper severe mosaic virus and comparison with other potyviruses. AB - The complete nucleotide sequence of pepper severe mosaic virus (PepSMV) was determined. The viral genome consisted of 9890 nucleotides, excluding a poly (A) tract at the 3' end of the genome. The PepSMV RNA genome encoded a single polyprotein of 3085 amino acid residues, resulting in ten functionally distinct potyviral proteins. The lengths of the 5' nontranslated region (NTR) and the 3' NTR were 164 and 468 nucleotides, respectively. The genome organization of the virus was typical for members of the genus Potyvirus in the family Potyviridae. The coat protein amino acid sequence identity between PepSMV and the other 45 potyviruses ranged from 53.4 to 79.7%. Sequence alignments and phylogenetic analyses of the potyviral polyprotein sequences revealed that PepSMV was the closest to potato virus Y (PVY) and closely related to members of the PVY subgroup. Our genome sequence data clearly confirmed that PepSMV belongs to a separate species in the genus Potyvirus. PMID- 16699830 TI - The role of the C-terminal region of olive latent virus 1 coat protein in host systemic infection. AB - A full-length cDNA clone of olive latent virus 1 (OLV-1), a member of the genus Necrovirus, family Tombusviridae, was subjected to site-directed mutagenesis, and coat protein gene mutants were constructed. A mutant clone, denoted Delta3297, was obtained by deleting the nucleotide in position 3297, thus inducing a frameshift and replacing the last 49 amino acids of the viral coat protein (CP) by a shorter sequence of 39 amino acids. This mutant was viable, stable, able to synthesize a smaller CP, and able to give rise to the formation of apparently intact virus particles. Cell-to-cell movement of Delta3297 in Nicotiana benthamiana leaves was not affected, but, contrary to wild type OLV-1, it failed to spread systemically. These results indicate that virion formation is necessary but not sufficient for long-distance movement for OLV-1 and highlights the role of the CP carboxy-terminal domain in systemic infection. PMID- 16699833 TI - Host-parasitoid dynamics of a generalized Thompson model. AB - A discrete-time host-parasitoid model including host-density dependence and a generalized Thompson escape function is analyzed. This model assumes that parasitoids are egg-limited but not search-limited, and is proven to exhibit five types of dynamics: host failure in which the host goes extinct in the parasitoid's presence or absence, unconditional parasitoid failure in which the parasitoid always goes extinct while the host persists, conditional parasitoid failure in the host and the parasitoid go extinct or coexist depending on the initial host-parasitoid ratio, parasitoid driven extinction in which the parasitoid invariably drives the host to extinction, and coexistence in which the host and parasitoid coexist about a global attractor. The latter two dynamics only occur when the parasitoid's maximal rate of growth exceeds the host's maximal rate of growth. Moreover, coexistence requires parasitism events to be sufficiently aggregated. Small additive noise is proven to alter the dynamical outcomes in two ways. The addition of noise to parasitoid driven extinction results in random outbreaks of the host and parasitoid with varying intensity. Additive noise converts conditional parasitoid failure to unconditional parasitoid failure. Implications for classical biological control are discussed. PMID- 16699834 TI - A simple model for the two dimensional blood flow in the collapse of veins. AB - Veins in the cardiovascular system may collapse if the internal pressure is less than the external pressure. Such collapse or buckling will have important consequence in terms of the rate of blood flow. Here a steady, parallel unidirectional flow as an exact solution of the continuity and the Navier Stokes equations is constructed. Various stages of the deformation process of the elastic tube (before contact of opposite sides occurring), from an ellipse to a ;strongly buckled' configuration, are obtained in analytical forms as a by product of the calculations. The pressure -- area and the pressure -- flow rate diagrams computed numerically from the model agree with the trends measured experimentally. PMID- 16699832 TI - Compliance and efficiency before and after implementation of a clinical practice guideline for laryngeal carcinomas. AB - We evaluated whether the implementation of a nationwide clinical practice guideline for diagnosis, treatment and follow-up of laryngeal carcinomas led to changes in hospital costs, balanced against clinical changes observed following the guideline's implementation. Charts of 822 patients with larynx carcinoma (459 treated before the introduction of the guideline and 363 thereafter) in five hospitals were retrospectively investigated. In all phases, no differences in total hospital costs were observed after the guideline's implementation. Total mean costs were Euro 3,207 (95%CI 3,091-3,395) for diagnosis, Euro 3,169 (2,153 4,182), Euro 5,026 (3,996-6,057), Euro 6,458 (5,579-7,337), Euro 8,037 (7,469 8,606), Euro 12,765 (10,763-14,769), Euro 19,227 (16,848-21,605) for treatment of dysplasia, carcinoma in situ, T1, T2, T3 and T4 carcinoma, respectively, and Euro 1,856 (1,491-2,220) for 1 year disease-free follow-up. In an earlier study, we observed several positive changes after the guideline's implementation. Balanced against the equal costs before and after the guideline's implementation, we conclude that the efficiency of the care process improved. PMID- 16699835 TI - Molecular cloning of the rat beta-carotene 15,15'-monooxygenase gene and its regulation by retinoic acid. AB - BACKGROUND: beta-Carotene exhibits biological activity as provitamin A. Key step in vitamin A formation is the cleavage of beta-carotene to retinal by an enzyme designated as beta-carotene 15,15'-monooxygenase (BCM). Recently, it is reported that expression of BCM gene in the intestine is under feedback regulation by retinoic acid (RA). However, the regulation of BCM gene expression in various other tissues is still unknown. AIM OF THE STUDY: In the present study, we identified the full-length cDNA encoding the rat BCM gene and investigated the regulation of its expression in several tissues by RA in the presence of vitamin A deficiency. METHODS: We cloned the full-length cDNA encoding BCM gene from a rat intestinal cDNA library by hybridization screening. The BCM gene expression was examined using Northern blotting and reverse transcription-PCR analysis. We also investigated whether BCM gene expression was regulated by retinoids in several tissues of vitamin A-deficient rats. RESULTS: Sequence analysis of this clone revealed an open reading frame of 1,701 bases encoding a protein of 566 amino acids. The predicted polypeptide showed 94%, 81%, and 66% identity with mouse, human, and chicken BCM, respectively. Rat BCM mRNA was highly expressed in the intestine and liver, while there was weak expression in the testes, kidneys, and lungs. Immunoblotting revealed that rat BCM is a 64-kDa protein. BCM gene expression was increased in the small intestine by vitamin A deficiency compared with that in rats on a control diet, while this upregulation was suppressed by all-trans RA (ATRA) or 9-cis RA (9-cis RA). BCM gene expression in the lungs and testes was also suppressed by ATRA or 9-cis RA in rats with vitamin A deficiency. However, hepatic BCM gene expression was only decreased by ATRA and renal expression was not affected by either retinoid. As the small intestine is the major site of beta-carotene conversion, intestinal BCM gene expression may be more tightly regulated. CONCLUSION: These data suggest that BCM gene expression in several tissues may be down-regulated by RA at the level of conversion of beta carotene to retinal. To prevent an excess of retinol, homeostasis may occur at the level of conversion of beta-carotene to retinal in several tissues. PMID- 16699836 TI - Effect of maternal exposure to homocystine on sodium valproate-induced neural tube defects in the mouse embryos. AB - BACKGROUND: Neural tube defects (NTD) are mainly of multifactorial origin. Maternal treatment with valproic acid (VPA) during pregnancy induces NTD in susceptible fetuses. Elevated levels of homocysteine are observed in pregnancies with NTD. The mechanism by which homocysteine might cause NTD is unknown. AIM OF THE STUDY: The aim of this study was to determine if homocystine would augment VPA-induced exencephaly in an experimental model. METHODS: Groups of mice were injected (IP) on gestational day 8 (GD) with a single dose of 75 mg/kg of L: Homocystine (HC) or a proportionate volume of saline, followed by a single dose of 600 mg/kg of VPA or an equal volume of saline. In a second experiment, mice were treated with a daily dose of 75 mg/kg of HC or an equal volume of saline (IP) from GD 5 and continued through GD 10. These animals had a single exposure to 600 mg/kg of VPA or saline (IP) on GD 8. All animals were killed by cervical dislocation on GD 18. Plasma homocysteine, folate and vitamin B12 were determined on GD 8 and GD 10 from single and multiple dose groups of mice, respectively, from additional experiments. RESULTS: The VPA and HC+VPA induced significantly higher rates of embryonic resorption and intrauterine growth retardation (IUGR) than HC or saline alone. HC + VPA groups had significantly more numerous fetuses with severe IUGR than HC alone or VPA alone groups. Both single and multiple doses of HC augmented VPA-induced reduction in fetal body weight. Successive doses of HC did not augment the rate of IUGR more significantly than a single dose of HC. Incidence of exencephaly was significantly enhanced in the HC + VPA groups compared to that in the HC or VPA alone groups. HC alone was not teratogenic. Plasma homocysteine levels increased several fold both in HC and HC + VPA groups and the increase was not particularly more marked in multiple dose groups than in the single dose groups. VPA did not elevate homocysteine concentration. Both FA and vitamin B12 concentrations were reduced by VPA, HC and HC + VPA, but HC and VPA when combined did not produce an additive effect on vitamin levels. CONCLUSION: These data indicate that HC and VPA interact in neurulation stage embryos, affect fundamental processes of closure of the neural tube and lead to enhanced incidence of NTD. PMID- 16699837 TI - Anxiety, obsessions and morbid preoccupations in pregnancy and the puerperium. AB - 129 mothers referred to specialist psychiatric services in Birmingham and Christchurch were interviewed with the Birmingham Interview. Anxiety disorders were more frequent than depression during pregnancy, and equally frequent after delivery. The focus of pre- and post-partum anxiety may be important for psychological treatment. At a severe level, the most common prepartum theme was fear of foetal death; this was associated with a history of reproductive losses or infertility. After delivery the commonest themes were the pathological fear of cot death and fear of the criticism of mothering skills (which was a clue to a disordered mother-infant relationship). Clinicians should be vigilant for obsessional disorders, querulant (complaining) disorders, post-traumatic stress disorder, conjugal jealousy and dysmorphophobic states, which are all quite common. Patients with "postpartum depression" usually had at least one other (co morbid) disorder, and 27% had two or more. These findings emphasize the diversity of postpartum psychiatric illness. PMID- 16699838 TI - Severe disorders of the mother-infant relationship: definitions and frequency. AB - We assessed the frequency of severe disorders of the mother-infant relationship in over 200 mothers referred to services in Birmingham and Christchurch, all of whom were interviewed using the Birmingham Interview for Maternal Mental Health. 10.6% had established rejection and 14.6% threatened rejection of their infants. 28.6% had various degrees of pathological anger, which was severe in 8.3%. Rejection was strongly associated with unwanted pregnancy and lack of interaction with the foetus. PMID- 16699839 TI - Dopamine modulation in the basal ganglia locks the gate to working memory. AB - The prefrontal cortex and basal ganglia are deeply implicated in working memory. Both structures are subject to dopaminergic neuromodulation in a way that exerts a critical influence on the proper operation of working memory. We present a novel network model to elucidate the role of phasic dopamine in the interaction of these two structures in initiating and maintaining mnemonic activity. We argue that neuromodulation plays a critical role in protecting memories against both internal and external sources of noise. Increases in cortical gain engendered by prefrontal dopamine release help make memories robust against external distraction, but do not offer protection against internal noise accompanying recurrent cortical activity. Rather, the output of the basal ganglia provides the gating function of stabilization against noise and distraction by enhancing select memories through targeted disinhibition of cortex. Dopamine in the basal ganglia effectively locks this gate by influencing the stability of up and down states in the striatum. Dopamine's involvement in affective processing endows this gating with specificity to motivational salience. We model a spatial working memory task and show that these combined effects of dopamine lead to superior performance. PMID- 16699840 TI - Propagation of photon noise and information transfer in visual motion detection. AB - The extraction of the direction of motion from the time varying retinal images is one of the most basic tasks any visual system is confronted with. However, retinal images are severely corrupted by photon noise, in particular at low light levels, thus limiting the performance of motion detection mechanisms of what sort so ever. Here, we study how photon noise propagates through an array of Reichardt type motion detectors that are commonly believed to underlie fly motion vision. We provide closed-form analytical expressions of the signal and noise spectra at the output of such a motion detector array. We find that Reichardt detectors reveal favorable noise suppression in the frequency range where most of the signal power resides. Most notably, due to inherent adaptive properties, the transmitted information about stimulus velocity remains nearly constant over a large range of velocity entropies. PMID- 16699841 TI - Factors affecting phase synchronization in integrate-and-fire oscillators. AB - Step changes in input current are known to induce partial phase synchrony in ensembles of leaky integrate-and-fire neurons operating in the oscillatory or "regular firing" regime. An analysis of this phenomenon in the absence of noise is presented based on the probability flux within an ensemble of generalized integrate-and-fire neurons. It is shown that the induction of phase synchrony by a step input can be determined by calculating the ratio of the voltage densities obtained from fully desynchronized ensembles firing at the pre and post-step firing rates. In the limit of low noise and in the absence of phase synchrony, the probability density as a function of voltage is inversely proportional to the time derivative along the voltage trajectory. It follows that the magnitude of phase synchronization depends on the degree to which a change in input leads to a uniform multiplication of the voltage derivative over the range from reset to spike threshold. This analysis is used to investigate several factors affecting phase synchronization including high firing rates, inputs modeled as conductances rather than currents, peri-threshold sodium currents, and spike-triggered potassium currents. Finally, we show that without noise, the equilibrium ensemble density is proportional to the phase response curve commonly used to analyze oscillatory systems. PMID- 16699842 TI - Mean-field analysis of selective persistent activity in presence of short-term synaptic depression. AB - Mean-Field theory is extended to recurrent networks of spiking neurons endowed with short-term depression (STD) of synaptic transmission. The extension involves the use of the distribution of interspike intervals of an integrate-and-fire neuron receiving a Gaussian current, with a given mean and variance, in input. This, in turn, is used to obtain an accurate estimate of the resulting postsynaptic current in presence of STD. The stationary states of the network are obtained requiring self-consistency for the currents-those driving the emission processes and those generated by the emitted spikes. The model network stores in the distribution of two-state efficacies of excitatory-to-excitatory synapses, a randomly composed set of external stimuli. The resulting synaptic structure allows the network to exhibit selective persistent activity for each stimulus in the set. Theory predicts the onset of selective persistent, or working memory (WM) activity upon varying the constitutive parameters (e.g. potentiated/depressed long-term efficacy ratio, parameters associated with STD), and provides the average emission rates in the various steady states. Theoretical estimates are in remarkably good agreement with data "recorded" in computer simulations of the microscopic model. PMID- 16699843 TI - Neural network model of the primary visual cortex: from functional architecture to lateral connectivity and back. AB - The role of intrinsic cortical dynamics is a debatable issue. A recent optical imaging study (Kenet et al., 2003) found that activity patterns similar to orientation maps (OMs), emerge in the primary visual cortex (V1) even in the absence of sensory input, suggesting an intrinsic mechanism of OM activation. To better understand these results and shed light on the intrinsic V1 processing, we suggest a neural network model in which OMs are encoded by the intrinsic lateral connections. The proposed connectivity pattern depends on the preferred orientation and, unlike previous models, on the degree of orientation selectivity of the interconnected neurons. We prove that the network has a ring attractor composed of an approximated version of the OMs. Consequently, OMs emerge spontaneously when the network is presented with an unstructured noisy input. Simulations show that the model can be applied to experimental data and generate realistic OMs. We study a variation of the model with spatially restricted connections, and show that it gives rise to states composed of several OMs. We hypothesize that these states can represent local properties of the visual scene. PMID- 16699847 TI - MR imaging of CNS tumors: are all contrast agents created the same? AB - The challenging and multiple goals and requirements of CNS tumor imaging require optimization of the CNS imaging protocols, including selection of the optimal contrast medium. There are several gadolinium compounds available for imaging the CNS, and all of these except the compound Gd-BOPTA have nearly identical profiles. The unique protein interaction of Gd-BOPTA results in double T1 and T2 relaxivity. This article summarizes the advantages of Gd-BOPTA over conventional gadolinium compounds in both conventional and functional MR imaging. The results of recent studies in adults and pediatric patients are presented and discussed. The principal conclusion of this article is that not all contrast agents approved for MR imaging of the CNS behave in the same way. The use of the high-relaxivity agent Gd-BOPTA for morphological and functional MR imaging allows improved detection and delineation of CNS lesions compared to conventional gadolinium agents. This not only permits more confident diagnoses, but also results in a substantially improved differential diagnostic process. The higher relaxivity of Gd-BOPTA helps to optimize functional MR imaging studies e.g. perfusion MR imaging and other dynamic MR imaging protocols. PMID- 16699848 TI - MR angiography of the carotid arteries and intracranial circulation: advantage of a high relaxivity contrast agent. AB - Several studies have shown the usefulness of contrast-enhanced MR angiography (CE MRA) for imaging the supraortic vessels, and, as a consequence, it has rapidly become a routine imaging modality. The main advantage over unenhanced techniques is the possibility to acquire larger volumes, allowing demonstration of the carotid artery from its origin to the intracranial portion. Most published studies on CE-MRA of the carotid arteries have been performed with standard Gd based chelates whose T1 relaxivity values are similar. Recently new gadolinium chelates such as gadobenate dimeglumine (Gd-BOP-TA, MultiHance; Bracco Imaging, Milan, Italy) have been developed which have markedly higher intravascular T1 relaxivity values. When administered at an equivalent dose to that of a standard agent, these newer contrast agents produce significantly greater intravascular signal enhancement. The availability of an appropriate high-relaxivity contrast agent might also help to overcome some of the intrinsic technical problems (e. g. those related to flow) that affect time-of-flight (TOF) and phase contrast (PC) MR angiography of the intracranial vasculature. To avoid the problem of superimposition of veins, ultrafast gradient echo MRA techniques with very short TR and TE have been developed. Although the precise sequence parameters vary between manufacturers, they are basically similar. The choice between performing a time-resolved or high spatial resolution CE-MRA examination depends upon the precise clinical application. The most common applications include the study of cerebral aneurysms, arteriovenous malformations, dural arteriovenous fistulas and dural venous diseases. PMID- 16699849 TI - Contrast-enhanced MR imaging of the spine: when, why and how? How to optimize contrast protocols in MR imaging of the spine. AB - The optimal protocols and the role of contrast agents in spinal MR imaging are controversial. Although the diagnosis of many common spinal diseases can be reliably achieved by means of unenhanced images, contrast use is often necessary to improve lesion detection and differential diagnosis. The heterogeneity of the different spinal compartments and the wide variety of spinal pathology require tailored imaging strategies. Thus, the rules to achieve optimization of contrast protocols for MR imaging of the spine are frequently very different to those for brain imaging, and depend on the location and site of origin of the lesions in a specific spinal compartment, on the findings of unenhanced imaging, and on the concomitant use of fat-suppression techniques. Furthermore, in most cases, the small size of the examined structures requires a meticulous technique, and the administration of a contrast agent with high relaxivity, such as MultiHance, is advisable to enable the detection of tiny areas of contrast enhancement. The applications and clinical utility of post-contrast MR imaging are discussed with regard to different spinal diseases. PMID- 16699850 TI - Diffusion-perfusion in intra-axial brain tumors with high relaxivity contrast agents. AB - A high relaxivity contrast agent is indicated for use in MRI of the central nervous system to visualize lesions with an abnormal blood-brain barrier (BBB) or abnormal vascularity of the brain. We evaluated MultiHance (gadobenate dimeglumine, Gd-BOPTA) on T2*-weighted perfusion imaging in 33 histologically proven intra-axial brain tumors. The higher T1 relaxivity, and therefore better contrast-enhanced T1 imaging led to significantly better tumor delineation. The higher T2 relaxivity allowed high quality T2* perfusion MRI and post processed rCBV maps, with a dose of 0.1 mmol/kg MultiHance. PMID- 16699852 TI - Acute necrotizing esophagitis: role of nonsteroidal anti-inflammatory drugs. AB - BACKGROUND: Acute necrotizing esophagitis (ANE) is a rarely described entity that is thought to be a cause of upper gastrointestinal (UGI) bleeding, The present study examined the incidence of ANE among patients with UGI bleeding, as well as the clinical features of ANE, and the coexisting illnesses and medication histories of ANE patients. METHODS: A retrospective analysis of clinical and endoscopic findings and the clinical course in 16 patients with ANE was carried out over a 3-year period. RESULTS: We observed 16 patients (6%) of ANE in 239 patients with UGI bleeding during the 3-year period. The average age of the patients was 62.5 years. The lesions predominantly affected the lower third of the esophagus, and hiatal hernia was the most common (63%) coexisting endoscopic finding. All patients had coexisting disease. Fifty percent of patients with ANE (eight patients) had taken nonsteroidal anti-inflammatory drugs (NSAIDs). ANE also occurred in four patients with diabetic ketoacidosis. Supportive therapy, including parenteral nutrition and administration of a proton pump inhibitor, was effective. CONCLUSIONS: ANE is more common than has been previously reported, and it should be included in the differential diagnosis of UGI bleeding. ANE could be characterized as an "acute esophageal mucosal lesion," particularly in aged patients with hiatal hernia and among those who consume NSAIDs. PMID- 16699851 TI - Colorectal cancer: genetics of development and metastasis. AB - It has been well documented that there are two major pathways in colorectal carcinogenesis. One is the chromosomal instability pathway (adenoma-carcinoma sequence), which is characterized by allelic losses on chromosome 5q (APC), 17p (p53), and 18q (DCC/SMAD4), and the other is a pathway that involves microsatellite instability. Recent progress in molecular biology, however, has shown that colorectal carcinogenesis is not necessarily clearly divided into these two pathways, but is in fact more complicated. Other routes, including the transforming growth factor-beta/SMAD pathway, the serrated pathway, and the epigenetic pathway, have been reported. Cross talk among these pathways has also been reported. In the invasion and metastasis steps of colorectal cancers, many more genes have now been identified as being involved in proteolysis, adhesion, angiogenesis, and cell growth. Recently accumulated evidence indicates that colorectal cancer is a genetically heterogeneous and complicated disease. PMID- 16699853 TI - Role of pancreatic trypsin in chronic esophagitis induced by gastroduodenal reflux in rats. AB - BACKGROUND: Reflux of the duodenal contents with gastric acid has been reported to contribute to the development of esophageal mucosal damage and inflammation. Recent studies show that pancreatic trypsin can stimulate the production of inflammatory mediators, including chemokines and prostaglandins from human esophageal epithelial cells in vitro. The aim of the present study was to investigate the role of pancreatic trypsin in the pathogenesis of chronic esophageal inflammation induced by gastroduodenal reflux in rats. METHODS: Esophagogastroduodenal anastomosis was carried out in male Wistar rats by anastomosing the jejunum to the gastroesophageal junction under diethyl ether inhalation anesthesia. The animals undergoing surgery were treated with the control diet, rabeprazole sodium, nizatidine, ecabet sodium, camostat mesilate (CMM), ONO-1714, a specific inducible nitric oxide synthase (iNOS) inhibitor, or meloxicam, a selective cyclooxygenase-2 (COX-2) inhibitor. Esophageal injury was evaluated by macroscopic and microscopic findings, and mRNA expression for CINC 1, COX-2, and iNOS was determined by real-time polymerase chain reaction (PCR). Trypsin activity within the esophageal lumen was measured 2 weeks after surgery, and the expression of protease-activated receptor (PAR)-1 and -2 was confirmed by reverse transcription (RT)-PCR. RESULTS: At 8 weeks after surgery, gastroduodenal reflux induced esophageal erosions and ulcer formation as well as marked thickening of the esophageal wall. Histological study showed an increase of thickness of the esophageal mucosa, hyperplasia of the epidermis and basal cells, ulcer formation, and marked infiltration of inflammatory cells. The macroscopic ulcer score and histological ulcer length were significantly reduced by treatment with rabeprazole or CMM but not by nizatidine or ecabet sodium, compared with each control. Rabeprazole, nizatidine, or ecabet sodium did not affect the severity of mucosal hyperplastic scores or histological parameters in esophagitis. In contrast, the CMM group showed a significant decrease in the mucosal hyperplastic and inflammatory scores. The enhanced expression of CINC-1, COX-2, and iNOS mRNA in the control group was also markedly inhibited in the CMM treated group. ONO-1714 or meloxicam treatment significantly reduced the macroscopic scores of ulcer and hyperplasia. The trypsin activity in the esophageal lumen was significantly increased in the control diet group, and this increase was significantly inhibited in the CMM-treated group. The expression of PAR-1 and -2 mRNA was confirmed in rat esophageal epithelium. CONCLUSIONS: With this model, we have demonstrated that CMM significantly reduces inflammation and hyperplasia in the esophageal mucosa. These results indicate that trypsin, which is primarily inhibited by CMM, plays an important role in the mucosal damage induced by gastroduodenal reflux and that it can be a therapeutic target in patients with gastroduodenal reflux esophagitis. PMID- 16699854 TI - Differential modulation in the functions of intestinal dendritic cells by long- and medium-chain fatty acids. AB - BACKGROUND: Although dendritic cells (DCs) play significant roles in intestinal immune responses, little is known regarding the direct effects of luminal foods on DC functions in the intestinal mucosa. In this study, we examined the effects of fatty acids (FAs) with various chain length on the phagocytic function, antigen presentation, and chemotaxis of intestinal DCs. METHODS: DCs obtained from the thoracic duct lymph of mesenteric lymphadenectomized rats were cultured with long [arachidonic acid (AA) or oleic acid] or medium (octanoic acid) chain FAs with interleukin-4 and granulocyte macrophage-colony stimulating factor. Tumor necrosis factor-alpha was added in the maturation group. Phagocytic function was examined by the intake of fluorescent microbeads. The expression of cell surface molecules was determined by immunocytochemistry or fluorescence activated cell sorting (FACS). Antigen presentation ability was evaluated by coincubating keyhole limpet hemocyanin (KLH)-sensitized spleen lymphocytes and KLH-pulsed DCs. Migratory ability of DCs toward the chemokines CC chemokine ligand (CCL) 20 and CCL21 was also assessed. RESULTS: There was a maturation induced decrease in phagocytic function, and an increased expression of major histocompatibility complex (MHC) class II molecules. Exposure of DCs to both long and medium-chain FAs maintained phagocytic ability. The expression of MHC class II molecules was significantly suppressed only by long-chain FAs. The expression of costimulatory factors was suppressed only by AA. Long- but not medium-chain FAs suppressed the antigen presentation ability of DCs induced by maturation. Chemotactic ability of mature DCs toward CCL21 was abrogated only by long-chain FAs. CONCLUSIONS: The data suggest that intraluminal exposure to long- and medium chain FAs may differentially modulate the immune functions of intestinal DCs. PMID- 16699855 TI - Repetitive rectal painful distention induces rectal hypersensitivity in patients with irritable bowel syndrome. AB - BACKGROUND: A reduced rectal perceptual threshold has been reported in patients with irritable bowel syndrome (IBS), but this phenomenon may be induced by a comorbid psychological state. We evaluated the rectal pain threshold at baseline and after conditioning (repetitive rectal painful distention: RRD) in patients with IBS or functional abdominal pain syndrome (FAPS), which is an abdominal pain disorder, and in healthy controls, and determined whether rectal hypersensitivity is a reliable marker for IBS. METHODS: The rectal sensory threshold was assessed by a barostat. First, a ramp distention of 40 ml/min was induced, and the threshold of pain and the maximum tolerable pressure (mmHg) were measured. Next, RRD (phasic distentions of 60-s duration separated by 30-s intervals) was given with a tracking method until the subjects had complained of pain six times. Finally, ramp distention was induced again, and the same parameters were measured. The normal value was defined by calculating the 95% confidence intervals of controls. RESULTS: Five or six of the seven IBS patients showed a reduced rectal pain threshold or maximum tolerable pressure, respectively, at baseline. In all patients with IBS, both thresholds were reduced after RRD load, but they were reduced in none of the patients with FAPS. RRD significantly reduced both thresholds in the IBS group (P < 0.05), but it had no effect in the control or FAPS groups. CONCLUSIONS: Rectal hypersensitivity induced by RRD may be a reliable marker for IBS. Conditioning-induced visceral hypersensitivity may play a pathophysiologic role in IBS. PMID- 16699856 TI - Molecular characterization and distribution of motilin family receptors in the human gastrointestinal tract. AB - BACKGROUND: Motilin and ghrelin have been recognized as important endogenous regulators of gastrointestinal motor function in mammals, mediated respectively by the motilin receptor and by the closely related ghrelin receptor. The aims of this study were to explore the distribution of motilin and ghrelin receptors along the human gastrointestinal tract and to establish the molecular nature of the human motilin receptor. METHODS: Post mortem and surgical human tissue specimens with no hemorrhage, necrosis, or tumor were obtained from various parts of the gastrointestinal tract. We analyzed levels of expression of mRNA for motilin and ghrelin receptors and examined their molecular identities. Portions of some specimens were also studied by immunohistochemistry for expression of the motilin and ghrelin receptor. RESULTS: The long form of the motilin receptor, but not the short form, was expressed in all parts of the gastrointestinal tract, and expressed at higher levels in muscle than in mucosa. Motilin receptor immunoreactivity was present in muscle cells and the myenteric plexus, but not in mucosal or submucosal cells. In contrast, ghrelin receptor mRNA was expressed equally in all parts of the gastrointestinal tract, with similar levels of expression in mucosal and muscle layers. CONCLUSIONS: Both the motilin and ghrelin receptors are expressed along the human gastrointestinal tract, but they have clearly distinct distributions in regard to both level and layer. The diffuse muscle expression of the motilin receptor, at both the levels of the gene and the protein product, along the entire gastrointestinal tract makes it a useful potential target for motilide drugs for dysmotility. PMID- 16699857 TI - Metabolism and hepatic toxicity of flutamide in cytochrome P450 1A2 knockout SV129 mice. AB - BACKGROUND: Flutamide, a nonsteroidal antiandrogen used for treatment of prostate cancer, causes a temporary increase in transaminase and in some cases severe liver dysfunction. It is dominantly metabolized by cytochrome P450 (CYP) 1A2 into 2-hydroxyflutamide (OH-flutamide), which has stronger antiandrogenic activity without obvious cytotoxicity to cultured hepatocytes. We hypothesized that another subsidiary metabolite might be responsible for induction of hepatotoxicity. METHODS: Flutamide was administered daily to CYP1A2 knockout mice and parental SV129 mice to compare pharmacokinetics and appearance of hepatic toxicity. RESULTS: In the CYP1A2 knockout mice, the plasma concentration of flutamide maintained at a high level and OH-flutamide stayed low; a higher amount of FLU-1, an alternative metabolite of flutamide, was detected in urine. Simple repetitive administration of 800 mg/kg of flutamide for 28 days to CYP1A2 knockout mice did not show abnormal elevation of plasma alanine aminotransferase (ALT). However, after the knockout mice were fed with an amino acid-deficient diet for 2 weeks, which reduced the glutathione (GSH) content to 27% of the initial, administration of 400 mg/kg of flutamide increased ALT to over 200 IU/l and histopathologically moderate hepatitis developed. Since FLU-1 itself did not show cytotoxicity or reduce GSH content in vitro, a further metabolized molecule must cause the hepatotoxicity. CONCLUSIONS: Blockade of CYP1A2 produced an unknown potential hepatotoxic molecule through FLU-1, and GSH might play an important role in diminishing the reactive hepatotoxic metabolite. PMID- 16699858 TI - High rates of progressive hepatic functional deterioration whether lamivudine therapy is continued or discontinued after emergence of a lamivudine-resistant mutant: a prospective randomized controlled study. AB - BACKGROUND: The management of patients with lamivudine-resistant mutants remains challenging, and no clear evidence has been presented concerning the discontinuation of lamivudine. METHODS: Seventy-four patients with lamivudine resistant mutants were prospectively enrolled and randomized; 37 patients continued (group A) and 37 patients discontinued lamivudine therapy (group B). The median follow-up was 20 months. RESULTS: Serum albumin levels were reduced and prothrombin time was prolonged in both groups versus baseline (P = 0.015 and 0.045, respectively). Four patients in group A (10.8%) and six in group B (16.2%) experienced hepatitis flare, but the difference was not significant (P > 0.05). Multivariate analyses identified a younger age as a risk factor for hepatitis flare (P = 0.021). Seven (18.9%) decompensations occurred in group A and five (13.5%) in group B, which was not a significant difference (P > 0.05). Multivariate analyses revealed higher alanine aminotransferase and a lower platelet count as risk factors for hepatic decompensation (P = 0.001 and 0.001, respectively). The patients whose platelet count was <65 000/microl experienced hepatic decompensations more frequently (50%) than those with platelet counts >65 000/microl (13.2%) during follow-up (P = 0.05). CONCLUSIONS: The clinical course of group B was not significantly different from that of group A. Therefore, the discontinuation of lamivudine may be a feasible option when other antiviral agents active against lamivudine-resistant mutants are unavailable. PMID- 16699859 TI - Modified Japan Integrated Staging is currently the best available staging system for hepatocellular carcinoma patients who have undergone hepatectomy. AB - BACKGROUND: We previously reported the effectiveness of the modified Cancer of the Liver Italian Program (CLIP) score in hepatocellular carcinoma (HCC) staging. To determine the best predictive staging system for HCC patients, we conducted a comparative analysis of prognosis using multivariate analysis in 230 Japanese HCC patients following hepatic resection. METHODS: We compared overall survival as predicted by different staging systems: the tumor node metastasis (TNM) system by the Liver Cancer Study Group of Japan, the Japan Integrated Staging (JIS) score (Japanese TNM and Child-Pugh classification), the modified JIS score using liver damage grade, the CLIP score, and our modified CLIP score using protein induced by vitamin K absence or the antagonist II (PIVKA-II). RESULTS: By a univariate analysis the PIVKA-II level (cut-off level, 400 mAU/ml) was significantly associated with patient survival (P = 0.031); however, alpha-fetoprotein level was not related to survival. Liver damage grade was significantly associated with patient survival (P = 0.039), although Child-Pugh classification was not related to survival. Univariate analysis showed that prediction of survival, according to disease stage, was better with the modified JIS score than with the TNM system, CLIP, modified CLIP, or JIS score. Multivariate analysis showed the modified JIS score showed the best ability to predict overall survival according to disease stage (Hazard ratio, 1.77; P = 0.002), and its Akaike information criteria statistic was the lowest (634.3). CONCLUSIONS: The modified JIS score, a staging system that combines tumor factors and hepatic function, is a better predictor of prognosis than other systems in HCC patients who have undergone hepatic resection. PMID- 16699860 TI - Hepatitis C virus core protein inhibits deoxycholic acid-mediated apoptosis despite generating mitochondrial reactive oxygen species. AB - BACKGROUND: Hepatitis C virus (HCV) core protein is known to cause oxidative stress and alter apoptosis pathways. However, the apoptosis results are inconsistent, and the real significance of oxidative stress is not well known. The aim of this study was twofold. First, we wanted to confirm whether core induced oxidative stress was really significant enough to cause DNA damage, and whether it induced cellular antioxidant responses. Second, we wanted to evaluate whether this core-induced oxidative stress and the antioxidant response to it was responsible for apoptosis changes. METHODS: HCV core protein was expressed under control of the Tet-Off promoter in Huh-7 cells and HeLa cells. We chose to use deoxycholic acid (DCA) as a model because it is known to produce both reactive oxygen species (ROS) and apoptosis. RESULTS: Core expression uniformly increased ROS and 8-hydroxy-2'-deoxyguanosine (8-OHdG) under basal and DCA-stimulated conditions. Core protein expression also increased manganese superoxide dismutase levels. Core protein inhibited DCA-mediated mitochondrial membrane depolarization and DCA-mediated activation of caspase-9 and caspase-3, despite the increase in ROS by DCA. Core protein inhibited DCA-mediated apoptosis by increasing Bcl-x(L) protein and decreasing Bax protein, without affecting the proportion of Bax between mitochondria and cytosol, resulting in suppression of cytochrome c release from mitochondria into cytoplasm. CONCLUSIONS: HCV core protein induces oxidative DNA damage, whereas it inhibits apoptosis that is accompanied by enhancement of ROS production. Thus, oxidative stress and apoptosis modulation by core protein are independent of each other. PMID- 16699862 TI - A solid pseudopapillary tumor arising from the greater omentum followed by multiple metastases with increasing malignant potential. AB - Solid pseudopapillary tumor is a rare, indolent neoplasm almost exclusively seen in the pancreas. We describe an unusual case of solid pseudopapillary tumor arising in the greater omentum of a 45-year-old man with subsequent multiple liver metastases and peritoneal dissemination. The patient underwent a total of ten laparotomies and died of unresectable disease 8 years after the initial presentation. Microscopically, the primary tumor and the relapsed tumors consistently exhibited identical growth patterns, which were characterized by solid sheets intermingling with pseudopapillary arrangements of uniformly small cells. Immunohistochemical staining was diffusely positive for vimentin and focally positive for alpha-1-antitrypsin. These features were compatible with those of conventional pancreatic solid pseudopapillary tumors. We also performed quantitative evaluation of Ki-67 immunoreactivity and mitotic figures, which indicated malignant transformation of this extremely rare tumor. This is the first detailed report of solid pseudopapillary tumor arising outside the pancreas complicated by repetitive liver metastases and peritoneal carcinomatosis, suggesting the existence of a more lethal subgroup of tumors. PMID- 16699863 TI - Multiple gastric ulcers caused by a rice cake as an intragastric foreign body. PMID- 16699861 TI - Promoter methylation profile in gallbladder cancer. AB - BACKGROUND: Methylation in the promoter region of genes is an important mechanism of inactivation of tumor suppressor genes. Our objective was to analyze the methylation pattern of some of the genes involved in carcinogenesis of the gallbladder, examining the immunohistochemical expression of proteins, clinical features, and patient survival time. METHODS: Twenty cases of gallbladder cancer were selected from the frozen tumor bank. The DNA extracted was analyzed by means of a methylation-specific polymerase chain reaction test for the CDKN2A (p16), MLH1, APC, FHIT, and CDH1 (E-cadherin) genes. Morphological and clinical data and follow-up information were obtained. RESULTS: All cases were in an advanced stage: histologically moderate or poorly differentiated tumors (95%). Methylation of the promoter area of genes was observed in 5%, 20%, 30%, 40%, and 65% of cases, and an altered immunohistochemical pattern (AIP) in 5%, 35%, 21%, 25%, and 66% for the MLH1, CDKN2A, FHIT, APC, and CDH1 genes, respectively. The Kappa concordance index between methylation of the promoter area and AIP for the MLH1 and CDH1 genes was very high (K > 0.75) and substantial for APC (K > 0.45). No correlation was found between survival time and the methylation of the genes studied. CONCLUSIONS: The high frequency of gene methylation (with the exception of MLH1) and the high agreement between AIP and methylation of the gene promoter area for the MLH1, APC, and CDH1 genes suggest that the inactivation of tumor suppressor genes and of the genes related to the control of cellular proliferation through this mechanism is involved in gallbladder carcinogenesis. PMID- 16699864 TI - Adefovir dipivoxil monotherapy is a sufficiently effective treatment modality for Japanese patients with lamivudine-resistant chronic hepatitis B. PMID- 16699865 TI - Treatment of proton pump inhibitor-resistant patients with gastroesophageal reflux disease. PMID- 16699866 TI - Do fatty acids influence functions of intestinal dendritic cells? PMID- 16699867 TI - What is the best staging system for hepatocellular carcinoma? PMID- 16699868 TI - Oxidative stress and apoptosis in hepatitis C: the core issue. PMID- 16699871 TI - Increased lipid peroxidation and ascorbic Acid utilization in testis and epididymis of rats chronically exposed to lead. AB - The hypothesis has been recently presented that lead may exert its negative effect at least partially through the increase of reactive oxygen species (ROS) level in tissues. However, little is known about the influence of lead intoxication on equilibrium between generation and elimination of ROS in the male reproductive system. Sexually mature male Wistar rats were given ad libitum 1% of aqueous solution of lead acetate (PbAc) for 9 months. Significantly higher lead concentrations were found in blood [median 7.03 (Q25-Q75: 2.99-7.65) versus 0.18 (0.12-0.99) microg dl-1, P < 0.01], caput epididymis [median 5.51 (Q25-Q75: 4.31 7.83) versus 0.51 (0.11-0.80) microg g-1 d.m., P < 0.001], cauda epididymis [median 5.88 (Q25-Q75: 4.06-8.37) versus 0.61 (0.2 - 1.08) microg g-1 d.m., P < 0.001] and testis [median 1.81 (Q25-Q75: 0.94-2.31) versus 0.17 (0.03-0.3) microg g-1 d.m., P < 0.01] of lead-intoxicated rats when compared to the control. The concentration of ascorbyl radical, generated in vitro from L: -ascorbic acid (present in tissues in vivo) was measured by means of Electron Paramagnetic Resonance (EPR) spectroscopy. The EPR signal of ascorbyl radical in caput epididymis, cauda epididymis, testis and liver of lead acetate-treated animals revealed a significant decrease by 53%, 45%, 40% and 69% versus control tissues, respectively. Plasma L: -ascorbic acid content measured by high performance liquid chromatography (HPLC) method and total antioxidant status (TAS) measured by means of spectrophotometry were also significantly lower in the intoxicated versus control animals (28% and 21%, respectively). In the group exposed to lead the concentration of lipid peroxide in homogenates of the reproductive system organs was significantly elevated versus control group. It can be assumed that the lower EPR signal was caused by decreased tissue concentrations of L: ascorbic acid. The latter may have resulted from consumption of ascorbic acid for scavenging of ROS excess in tissues of animals chronically exposed to lead. PMID- 16699873 TI - Relapse of primary central nervous system lymphoma: clinical features, outcome and prognostic factors. AB - Data on relapsed primary central nervous system lymphoma (PCNSL) are limited. We have evaluated the clinical characteristics and outcome of relapsed PCNSL patients from two German trials. Patients with relapsed disease after primary treatment were studied. Primary therapy consisted of high-dose methotrexate-based chemotherapy in all patients. Treatment for relapse was not predetermined. After a median follow-up of 22.5 months, 52 (36%) patients with relapse were identified among 143 patients with complete remission (CR) after primary treatment. The median disease-free survival was 10.25 (3-47.5) months. The median age at relapse was 59 years. Forty-four of 51 evaluable patients relapsed within the CNS, 6 systemically and one both cerebrally and systemically. The median survival time after first relapse was 4.5 (0.5-40.5) months. Karnofsky performance status (KPS) at relapse (P = 0.004), site of relapse (isolated systemic versus other, P = 0.049) and treatment for relapse (versus no treatment, P = 0.001) were independent prognostic factors for survival after relapse in multivariate analysis. Survival of patients with relapsed PCNSL is poor despite high response rates to salvage therapy. Good KPS, isolated systemic relapse and treatment for relapse were significantly associated with longer survival. PMID- 16699874 TI - Intraventricular solitary fibrous tumor: a rare location for a rare tumor. PMID- 16699877 TI - Depression and consumption: psychopharmaceuticals, branding, and new identity practices. AB - As pharmaceuticals are moving from private patient-doctor conversations to public television and print advertisements, best-selling books, and top TV shows, as well as into everyday conversations around risk and illness, how people understand health, sickness, and their own identity is also changing. This paper explores some of these changes by unpacking some of the social, political, and personal layers that are complicating the production and marketing of prescription drugs, and that are transforming the identity practices around contested illness. I focus on Sarafem and premenstrual dysphoric disorder (the illness Sarafem was marketed for) as a case study. PMID- 16699878 TI - Implication of cannabinoids in neurological diseases. AB - 1. Preparations from Cannabis sativa (marijuana) have been used for many centuries both medicinally and recreationally. 2. Recent advances in the knowledge of its pharmacological and chemical properties in the organism, mainly due to Delta(9)-tetrahydrocannabinol, and the physiological roles played by the endocannabinoids have opened up new strategies in the treatment of neurological and psychiatric diseases. 3. Potential therapeutic uses of cannabinoid receptor agonists include the management of spasticity and tremor in multiple sclerosis/spinal cord injury, pain, inflammatory disorders, glaucoma, bronchial asthma, cancer, and vasodilation that accompanies advanced cirrhosis. CB(1) receptor antagonists have therapeutic potential in Parkinson's disease. 4. Dr. Julius Axelrod also contributed in studies on the neuroprotective actions of cannabinoids. PMID- 16699900 TI - Safe psoriasis control: a new outcome measure for the composite assessment of the efficacy and safety of psoriasis treatment. AB - BACKGROUND: PASI is an inadequate outcome measure for the assessment of psoriasis treatments. No currently used endpoints provide a benefit: risk assessment of treatment taking into consideration all available efficacy and safety data. OBJECTIVE: To propose a new outcome measure called "safe psoriasis control" (SPC), which assesses multiple dimensions of the disease in a clinically meaningful way through the combined use of appropriate efficacy, quality of life, and safety data. METHODS: Data from 3,500 subjects were used for the purpose of derivation and validation of the SPC endpoint. Advanced statistical methodology was used to evaluate and validate important components in the assessment of therapeutic benefit. RESULTS: SPC was shown to be a simple but meaningful combined endpoint showing the proportion of patients who had treatment benefit without major side effects. CONCLUSION: The SPC endpoint may be a step-forward in providing a composite tool for the evaluation of treatments for psoriasis. PMID- 16699879 TI - Microarray analysis of gene expression in the supraoptic nucleus of normoosmotic and hypoosmotic rats. AB - 1. Hypoosmolality produces a dramatic inhibition of vasopressin (VP) and oxytocin (OT) gene expression in the supraoptic nucleus (SON). This study examines the effect of sustained hypoosmolality on global gene expression in the OT and VP magnocellular neurons (MCNs) of the hypothalamo-neurohypophysial system (HNS), in order to detect novel genes in this system that might be involved in osmoregulation in the MCNs. 2. For this purpose, we used Affymetrix oligonucleotide arrays to analyze the expression of specific genes in laser microdissected rat SONs, and their changes in expression during chronic hypoosmolality. We identified over 40 genes that had three-fold or more greater expression in the SON versus total hypothalamus, and that also changed more than two fold in expression as a result of the chronic hypoosmolar treatment. These genes contained both novel as well as genes previously known to be present in the SON. All of the raw data for the genes that are expressed in the SON and altered by hypoosmolality can be found on the following NINDS website URL address: http://data.ninds.nih.gov/Gainer/Publications. PMID- 16699901 TI - The Cochrane Skin Group: promoting the best evidence. AB - BACKGROUND: The international Cochrane Skin group, established in 1997, organizes, writes and disseminates systematic reviews of therapeutic interventions in dermatology. OBJECTIVE: To introduce the Cochrane Skin Group and what it offers to cutaneous medicine and surgery providers. METHODS: Descriptive review of the structure and output of the Cochrane Collaboration Library and the Cochrane Skin Group. RESULTS: Systematic reviews of randomized controlled trials produced through the Cochrane Skin Group provide a benchmark for evidence summaries for informing clinical decisions in dermatology. CONCLUSION: The work performed by the Cochrane Skin Group is an important component for informing the evidence base for the clinical practice of cutaneous and surgical dermatology. PMID- 16699902 TI - Unilateral Basal cell carcinomas: an unusual entity treated with photodynamic therapy. AB - BACKGROUND: Unilateral localized basal cell carcinomas are an uncommon finding that presents both a diagnostic and therapeutic challenge. Exclusion of unilateral nevoid basal cell carcinoma syndrome is indicated. There are few reports in the literature regarding this entity and even less regarding therapeutic strategies. OBJECTIVE: We present a patient with unilateral localized basal cell carcinomas who was successfully treated with photodynamic therapy. METHODS: Photodynamic therapy was started using Levulan) Kerastick) as previously described. The topical solution was applied to the patient's back and illuminated the following day via the BLU-U Blue Light Illuminator. RESULTS: The patient tolerated the procedure well and without complications. The patient had an excellent therapeutic response with no clinically apparent basal cell carcinomas for 18 months. CONCLUSIONS: We report a patient with unilateral basal cell carcinomas successfully treated with photodynamic therapy. This uncommon entity represents a diagnostic challenge in its inherent absence of the classic clinical and radiographic findings of nevoid basal cell carcinoma syndrome. Like nevoid basal cell carcinoma syndrome, unilateral basal cell carcinomas poses a therapeutic challenge with the sheer number of cutaneous tumors. The use of photodynamic therapy carries a proven therapeutic efficacy, a low rate of adverse events and excellent cosmesis. PMID- 16699903 TI - Safety of efalizumab in patients with moderate to severe chronic plaque psoriasis: review of clinical data. part II. AB - BACKGROUND: The efficacy and safety of efalizumab have been evaluated in multiple clinical trials. OBJECTIVE: The purpose of this review is to provide an overview of the safety profile of efalizumab during the clinical trials. METHODS: Twelve week data from four placebo-controlled trials were pooled and analyzed. Data from patients receiving 13-60 weeks of efalizumab therapy were pooled to evaluate longer-term safety. RESULTS: The most common adverse events were mild to moderate, self-limiting, flu-like symptoms that were most frequent following the first two efalizumab doses; by the third dose the incidence was comparable to placebo. Serious adverse events were observed in 2.2% and 1.7% of efalizumab- and placebo-treated patients, respectively. Nonserious adverse events leading to withdrawal were infrequent and similar to placebo (2.8% vs 1.8%). There does not appear to be increased risk of end-organ toxicity, infection, or malignancy in efalizumab-treated patients. CONCLUSION: Efalizumab was well tolerated, with a favorable safety profile. PMID- 16699904 TI - Clinical efficacy of efalizumab in patients with chronic plaque psoriasis: results from three randomized placebo-controlled Phase III trials: part I. AB - BACKGROUND: Effective psoriasis therapies are needed for long-term symptom control. OBJECTIVE: Assess efalizumab (Raptiva) efficacy in a large cohort of psoriasis patients. METHODS: Data from three Phase III, randomized, double-blind, parallel-group, placebo-controlled, multicenter studies were pooled. Patients (n = 1,651) with moderate to severe plaque psoriasis received 12 weeks of subcutaneous efalizumab 1 or 2 mg/kg/wk or placebo. RESULTS: All efficacy measures reached statistical significance within each of the individual studies (p < 0.001) and overall. More efalizumab-treated patients achieved > or = 75% and > or = 50% Psoriasis Area and Severity Index (PASI) improvement at week 12 than did placebo-treated patients (27.8% vs 3.8% [p < 0.001] and 56.1% vs 14.6% [p < 0.001], respectively). Significant PASI improvements occurred as early as week 2 (12.5% vs 7.9%, p =0.0001). Adverse events were generally mild to moderate. CONCLUSION: Efalizumab resulted in early and significant improvement for all efficacy endpoints and was well tolerated in patients with moderate to severe chronic plaque psoriasis. PMID- 16699905 TI - Multiple cutaneous and uterine leiomyomas associated with gastric GIST. AB - BACKGROUND: There are a number of reports documenting familial cases of leiomyomatosis cutis associated with uterine leiomyomata. However, to our knowledge, the association of gastrointestinal stromal tumour (GIST) with this entity has not as yet been reported. We report an interesting case of cutaneous leiomyomatosis, metachronous uterine leiomyomata, and a gastric GIST in a 43-year old woman. OBSERVATION: The patient had previously undergone two separate uterine myomectomies at ages 25 and 26 years, respectively, followed by a hysterectomy at 27 years. At 36 years she underwent partial gastrectomy for excision of GIST and this was followed by the development of extensive, symptomatic cutaneous leiomyomata at 43 years. In the report, we have documented histological, immunohistochemical and clinical observations and furthermore report on the therapeutic measures undertaken. CONCLUSION: We report an interesting association of cutaneous leiomyomatosis, uterine leiomyomas and GIST. PMID- 16699906 TI - Off-label dermatologic uses of anti-TNF-a therapies. AB - BACKGROUND: Tumor necrosis factor-alpha (TNF-a) is a proinflammatory cytokine that plays an immunomodulatory role in a variety of systemic and dermatologic diseases. Currently, three anti-TNF-a drugs are available in North America- infliximab (approved in the U.S. for the treatment of rheumatoid arthritis, Crohn's disease, ankylosing spondylitis, ulcerative colitis, and psoriatic arthritis), etanercept (approved in the U.S. for the treatment of rheumatoid arthritis, juvenile rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, and psoriasis), and adalimumab (approved for the treatment of rheumatoid arthritis and psoriatic arthritis). OBJECTIVE: To review the current literature supporting alternative (and currently off-label) dermatologic uses of TNF-a antagonists. METHODS: A MEDLINE search (1966-March 2005) was conducted using the keywords "infliximab," "etanercept," "adalimumab," "TNF inhibitors," and "off-label" to identify published reports of off-label dermatologic uses of TNF-a inhibitors. RESULTS: Anti-TNF-a therapies have been reported in the following dermatologic diseases: sarcoidosis, hidradenitis suppuritiva, cicatricial pemphigoid, Behcet's disease, pyoderma gangrenosum, multicentric reticulohistiocytosis, apthous stomatitis, Sneddon-Wilkinson disease, SAPHO syndrome, pityriasis rubra pilaris, eosinophilic fasciitis, panniculitis, Crohn's disease, necrobiosis lipoidica diabeticorum, dermatomyositis, and scleroderma. The vast majority of these reports are in the form of individual case reports and small case series. Only two published randomized controlled trials involving the off-label use of a TNF inhibitor were found. CONCLUSIONS: A growing number of published reports suggest that anti-TNF-a therapies may be effective in the treatment of numerous inflammatory skin diseases outside their currently approved indications. PMID- 16699907 TI - Molecular aspects of ultraviolet radiation-induced apoptosis in the skin. AB - BACKGROUND: Apoptosis, or programmed cell death, is an essential physiological process that controls cell numbers during physiological processes, and eliminates abnormal cells that can potentially harm an organism. OBJECTIVE: This review summarizes our current state of knowledge of apoptosis induction in skin by UV radiation. METHODS: A review of the literature was undertaken focusing on cell death in the skin secondary to UV radiation. RESULTS: It is evident that a number of apoptotic pathways, both intrinsic and extrinsic, are induced following exposure to damaging UV radiation. CONCLUSION: Although our understanding of the apoptotic processes is gradually increasing, many important aspects remain obscure. These include interconnections between pathways, wavelength-specific differences and cell type differences. PMID- 16699908 TI - Safety and efficacy of a milk-derived extract in the treatment of plaque psoriasis: an open-label study. AB - BACKGROUND: XP-828L is a nutraceutical compound obtained by the extraction of a growth factors-enriched protein fraction from bovine milk. XP-828L may improve psoriasis. OBJECTIVES: An open-label study was performed to determine the efficacy, tolerability and safety of XP-828L in the treatment of plaque psoriasis. METHODS: Eleven adult patients with chronic, stable plaque psoriasis on 2% or more of body surface area (BSA) received 5 g of oral XP-828L twice daily for 56 days. RESULTS: All 11 patients completed the 56 days of treatment. At day 28, 6 of the 11 patients showed a reduction in PASI score. At 56 days, seven subjects had a decrease in PASI score ranging from 9.5% to 81.3%. Eight (8) out of 11 patients agreed to participate in an additional 8-week extension treatment phase. Improvement of psoriasis was maintained during the extension period. No clinically significant adverse events or laboratory abnormalities occurred. CONCLUSION: XP-828L may improve psoriasis in patients with mild-to-moderate psoriasis. PMID- 16699909 TI - Misnomers in dermatology. AB - BACKGROUND: Dermatology lexicon is rich with descriptive terminology. However, for a variety of reasons, it also includes a number of misnomers. OBJECTIVE: To review the more commonly encountered and critically appraised misnomers in dermatology. METHODS: A search of MEDLINE (1966 - 2004), eMedicine dermatology text and electronic versions of two standard dermatology texts, Fitzpatrick's Dermatology in General Medicine and Dermatology, was performed using the permutations of the terms: dermatology, skin, cutaneous, and misnomer. RESULTS: Greater than 40 misnomers were identified. CONCLUSIONS: Misnomers in dermatology stem largely from lack of appreciation of underlying etiology or histopathological features of certain skin conditions, imprecise historical observations and erroneous eponymous credit. Historical, clinical, or histopathological explanations are used to clarify the nature of the misnomers, and in some cases suggestions for improved terminology are provided. PMID- 16699911 TI - Relationships between stand composition and ectomycorrhizal community structure in boreal mixed-wood forests. AB - We investigated the community structure of ectomycorrhizal fungi under varying overstory tree compositions in the southern mixed-wood boreal forest of Quebec. Sampling took place at two locations of differing postfire ages and nine 100-m2 plots were sampled per location. The dominant overstory tree species in the plots were trembling aspen (Populus tremuloides Michx.), white birch (Betula papyrifera Marsh.) or white spruce [Picea glauca (Moench) Voss], and balsam fir [Abies balsamea (L.) Mill.]. Mycorrhizae were analyzed using morphological as well as molecular methods, employing fungal-specific primers to amplify ribosomal DNA for subsequent cloning and sequencing. A total of 1800 mycorrhizal root tips collected from the 18 plots were morphologically classified into 26 morphotypes, with Cenococcum geophilum dominating (36% of root tips). A second set of root tips, selected from the same 18 samples on which the morphological analysis was based, were analyzed using molecular methods. From this analysis, 576 cloned polymerase chain reaction products were screened by restriction fragment length polymorphism analysis and a total of 207 unique types were found. No one type dominated the system and 159 occurred only once. Sequence analysis of the types that occurred more than once revealed that Piloderma sp., Russula sp., Cortinarius sp., and Lactarius sp. were the most common mycorrhizae. The ectomycorrhizal fungal community structure revealed by the rDNA analysis differed from that observed using morphological methods. Canonical correspondence analyses of the sequenced restriction types and % overstory composition indicate that the distributions of ectomycorrhizal fungi are influenced by the relative proportions of host tree species. The distinct fungal assemblages found in the different plots supported by the different combinations of host tree species provides further support for the need to conserve stand diversity in the southern boreal forest. PMID- 16699912 TI - Influence of different power densities of LILT on cultured human fibroblast growth : a pilot study. AB - The aim of this in vitro study was to analyze the influence of different power densities of low-intensity laser therapy (LILT) on the biomodulation of human gingival fibroblasts. The cells were cultured in nutritional deficit. Laser irradiation was carried out with a GaAlAs diode laser (lambda-660 nm, 2 J/cm2). The irradiation was done twice with 12-h interval using the punctual technique, at continuous mode and in contact. The experimental groups were: I, control, nonirradiated; II, power of 10 mW and intensity approximately or equal to 142.85 mW/cm2; III, 29 mW and 428.57 mW/cm2. Growth curves were obtained by using the trypan blue dye exclusion assay. The cell growth of the irradiated groups was significantly higher than control group (P or = 580 mg/m(2)/wk was considered successful. RESULTS: Thirty-nine patients (34 NSCLC, 5 epithelial neoplasias) were enrolled. SWOG grade 3-4 neutropenia and thrombocytopenia were observed in 17.9% and 12.8% of patients, respectively. Nonhematological toxicity was minimal. Twenty-eight (18%) of 158 cycles required dose modifications and/or delays. Twenty-five patients received a gemcitabine dose intensity of > or = 580 mg/m(2)/wk. The received DIs were 601.8 mg/m(2)/wk for gemcitabine and 21.0 for cisplatin, with a relative DIs of 90.3% and 90.1%, respectively. The response rate of 27 evaluable patients with NSCLC was 44% (95% confidence interval [CI], 25.3 to 62.7%). CONCLUSIONS: The shorter schedule of gemcitabine on days 1 and 4 plus cisplatin on day 2 produces an effective DI and a toxicity profile comparable to that of weekly regimens. PMID- 16699976 TI - Diagnosis of ischaemic heart disease by myocardial contrast echocardiography during supine bicycle stress. AB - INTRODUCTION: Myocardial contrast echocardiography (MCE) is a new imaging modality for diagnosing ischaemic heart disease (IHD). AIM: The aim of this study was to assess 1) the feasibility of MCE during supine bicycle stress and 2) the value of this method in the diagnosis of IHD. METHODS: Supine bicycle stress was performed in 44 consecutive patients (pts) referred for coronary angiography with an intermediate pre-test probability of IHD. MCE was carried out at peak stress and during recovery (once the heart rate returned to the pre-exercise value). During MCE an ultrasound contrast agent (Sonovue) was administered intravenously in a continuous mode using an infusion pump (BR-INF 100, Bracco Research). The acquired images were qualitatively assessed for perfusion and wall motion abnormalities. The 18-segment division of the left ventricle was used in the analysis. Coronary angiography was performed in all pts within 15 days of the exercise test. A quantitative analysis of coronary artery stenoses was carried out using the CAAS system. RESULTS: MCE could not be performed in 2 pts due to technical difficulties. Coronary angiography revealed significant coronary artery stenosis in 25 pts. The sensitivity and specificity of MCE in the diagnosis of IHD were 92.0% and 82.4%, respectively. The positive and negative predictive values were found to be 88.5% and 87.5%, respectively, while the agreement between coronary angiography and MCE was 88.1% (kappa=0.75). CONCLUSIONS: MCE during supine bicycle stress and continuous intravenous administration of an ultrasound contrast agent is a feasible technique and allows accurate diagnosis of IHD in pts in whom the pre-test probability of the disease is intermediate. PMID- 16699978 TI - Changes in C-reactive protein levels following coronary stent implantation depend on the extent of periprocedural arterial injury. AB - BACKGROUND: Coronary stenting is associated with acute inflammation within the arterial wall followed by neointimal growth. Acute inflammatory response is expressed by a marked systemic elevation of the inflammatory biomarker C-reactive protein (CRP). It has been shown that the degree and extent of CRP increase may be related to clinical presentation or periprocedural treatment. AIM: To investigate whether an increase in CRP level is associated with the extent of arterial injury during stent deployment. METHODS: CRP levels were measured with a high-sensitivity CRP (hsCRP) assay. Seventy-three patients (51 males) with normal baseline plasma CRP (<3 mg/L) underwent percutaneous coronary intervention (PCI) with stent implantation for stable coronary disease. Blood samples for hsCRP were drawn before the intervention, and 6, 12 and 24 hours after the procedure. Both quantitative (single vs multivessel coronary intervention) and qualitative analyses (including lesion classification according to the ACC/AHA grading system) were performed in all patients. The examined angiographic parameters and procedural data obtained included reference diameter, lesion location, lesion length, total stented segment length, diameter of stent after deployment, maximal deployment pressure, total inflation number and duration. RESULTS: The mean hsCRP level increased in all patients between baseline and 24 hours (1.36+/-0.93 mg/L and 4.34+/-3.3 mg/L, p <0.0001). Single vessel procedure was performed in 51 patients and multivessel coronary intervention in 22 patients. Mean hsCRP was similar at baseline and after 6 hours in both groups and the increase after 12 and 24 hours was higher among patients with multivessel coronary intervention compared to patients with single vessel procedure (2.69+/-2.48 vs 4.15+/-3.17; p=0.039 and 3.76+/-3.13 vs 5.69+/-3.38; p=0.021, respectively). There was no correlation between hsCRP and the degree of lesion complexity. Multiple regression analysis showed that the total stented segment length (p=0.01) contributed to the hsCRP increase after 24 hours. CONCLUSIONS: The inflammatory response expressed by hsCRP levels is higher in patients with multivessel coronary intervention with longer total segment stented. PMID- 16699980 TI - Evaluation of the defibrillation threshold in atrial fibrillation by transoesophageal cardioversion using a biphasic impulse. AB - INTRODUCTION: Recurrent atrial fibrillation (AF) in the setting of haemodynamic disturbances requires frequently repeated cardioversions, which is associated with the risk of myocardial damage. It is thus necessary to identify methods which can minimise the cardioverter impulse energy. AIM: To define the defibrillation threshold in recent-onset AF using a biphasic impulse, following an infusion of magnesium, potassium, and amiodarone. METHODS: Transoesophageal cardioversion was performed in 32 patients with AF lasting < or =48 hours, in whom prior administration of 40 mEq K+, 4.0 g MgSO4 and 300 mg amiodarone did not restore sinus rhythm. Cardioversion was performed under short intravenous anaesthesia using a biphasic impulse travelling from a multi-annular oesophageal electrode to two electrodes on the anterior chest wall. The initial energy was set to 1 J, which was subsequently increased according to the following protocol: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 40, 50 and 70 J. RESULTS: Electrical cardioversion following the administration of electrolytes and amiodarone restored sinus rhythm in all the patients (100% efficacy). The mean defibrillation threshold was 12.9+/-14.3 J, with a minimal effective energy of 1 J and a maximum effective energy of 70 J. The defibrillation threshold was in the range from 1 to 10 J in 75% of the patients. The mean cumulative energy transferred between electrodes during evaluation of the defibrillation threshold was 39.7 J (SD, 38.8). CONCLUSIONS: Transoesophageal cardioversion using a low energy (mean, 12.9 J; range, 1-70 J) biphasic impulse, following the intravenous administration of potassium chloride and amiodarone, was 100% effective in restoring sinus rhythm in AF. PMID- 16699982 TI - Relationship between tissue reperfusion and postinfarction left ventricular remodelling in patients with anterior wall myocardial infarction treated with primary coronary angioplasty. AB - INTRODUCTION: Pathological left ventricular remodelling is considered the main cause of heart failure in patients after myocardial infarction. AIM: The purpose of this study was to evaluate correlations between the degree of coronary microvascular reperfusion assessed by means of the angiographic myocardial blush grade (MBG) scale and adverse left ventricular remodelling in patients with acute myocardial infarction treated with primary coronary angioplasty. METHODS: This study involved 92 consecutive patients, hospitalised because of their first anterior wall myocardial infarction, who underwent successful (TIMI-3 grade flow) primary coronary angioplasty. Angiographic myocardial reperfusion parameters (MBG, corrected TIMI Frame Count) were assessed. Three days and 6 months after the index PCI all patients underwent an echocardiographic examination and such parameters as end-diastolic volume (EDV), left ventricular ejection fraction (EF) and contractility index (WMSI) were calculated. RESULTS: The patients were divided into two groups: group 1 with impaired myocardial reperfusion (MBG 0-1) (n=32) and group 2 with adequate tissue reperfusion (MBG 2-3) (n=60). Negative left ventricular remodelling was observed more frequently in group 1 than in group 2 (28.1% vs 10%, p=0.029). More patients in group 1 presented heart failure symptoms (56.3% vs 25%, p=0.013). CONCLUSIONS: Failure of tissue reperfusion assessed by means of angiographic indices (MBG 0-1) in patients with myocardial infarction treated with primary coronary angioplasty is associated with a higher rate of adverse myocardial remodelling and heart failure at 6 months after myocardial infarction. PMID- 16699984 TI - Elderly and patients with sick sinus syndrome have lower chances for appropriate pacemaker mode selection, according to the Polish Cardiac Society recommendations -- a single-centre retrospective analysis. AB - BACKGROUND: The Polish Cardiac Society recommendations for permanent heart pacing have been valid since 1999. The clinical use of these guidelines is, however, still limited. AIM: To analyse whether the chosen pacing strategy is consistent with the Polish Cardiac Society recommendations and to estimate the effects of analysed factors on selecting optimal or suboptimal pacing modes. METHOD: Retrospective analysis of medical records and procedure protocols of 1052 patients who underwent pacemaker implantation between 1 January 2000 and 31 December 2004 was performed. In each case, the applied pacing mode was compared against the optimal one defined according to the guidelines of the Polish Cardiac Society. A number of demographic and clinical factors associated with the procedure were analysed and correlated with the optimal pacing mode selection. RESULTS: During the analysed period, 59.3% of patients received optimal pacing. The percentage of patients with optimal pacing increased in the consecutive years from 40.2% in 2000 to 68.5% in 2005. In a univariate regression analysis, patients above the age of 70 years, with sick sinus syndrome as an indication for pacing, as well as cardiac heart failure and obesity, received optimal pacing significantly less frequently. In a multivariate analysis, advanced age and sick sinus syndrome were found to be independent predictors of suboptimal pacing. CONCLUSIONS: About 60% of patients had their pacemakers implanted with the optimal pacing mode selection according to the valid recommendations. Patients over the age of 70 years, as well as patients with sick sinus syndrome, had significantly lower chances of receiving optimal pacing. PMID- 16699988 TI - [Massive pulmonary embolism in a patient with ulcerative colitis and hyperhomocysteinaemia -- a case report]. AB - We describe a case of a 37-year-old man with active ulcerative colitis complicated by proximal deep vein thrombosis of the left lower limb and subsequent massive pulmonary embolism requiring mechanical ventilation and catecholamine infusion. In spiral CT a large thrombus obturating left pulmonary artery as well as bilateral embolic material in lobar and segmental branches were visible. Haemodynamic status improved after infusion of rtPA. Haemoglobin decrease (7.0-5.6 mmol/L) was corrected with erythrocyte mass transfusion. During subsequent therapy with intravenous full dose of unfractionated heparin and further long-term treatment with subcutaneous enoxaparin (1.5 mg/kg and after 3 months 1.0 mg/kg daily) haemoglobin value was relatively stable. Underlying disease was treated with 5-ASA (mesalazine) and steroids. Due to hyperhomocysteinaemia (16.0 micromol/L) coexisting with a low plasma folic acid (2.1 ng/ml) and cyanocobalamin (137 pg/ml) levels, supplementation with these vitamins was prescribed. The screening tests for familial thrombophilia (including 677C-->T MTHFR mutation) were negative. The authors discuss the pathogenesis of increased thromboembolic risk in inflammatory bowel disease and therapeutic dilemmas connected with treatment of such complications. PMID- 16699986 TI - Cytochrome P450 2C9 polymorphism and acenocoumarol therapy. AB - BACKGROUND: Oral anticoagulants, in Hungary acenocoumarol being the one exclusively used, have a low therapeutic index and a high bleeding complication rate. The cytochrome P450 2C9 enzyme plays an important role in their metabolism. AIM: To investigate the influence of CYP2C9 polymorphism on the occurrence of bleeding complications related to acenocoumarol therapy. METHODS: Genotyping of 421 patients (183 men and 238 women, mean age 66.2+/-11.8 years), who had been taking acenocoumarol for at least 6 months, was performed. Based on patient history and laboratory data, the correlations between genotype and acenocoumarol dose and bleeding complications were retrospectively analysed. RESULTS: In 145 patients bearing alleles with reduced activity (CYP2C9*2 and/or *3), the optimal dose of acenocoumarol was significantly (p<0.001) lower than in patients with the wild type allele (2.12+/-0.96 mg/day and 2.90+/-1.46 mg/day, respectively). In comparison with wild type allele patients, the mean daily acenocoumarol dose was lower in the CYP2C9*2 group, and the lowest in *3 bearers. Although the occurrence of minor bleeding complications in patients with the variant allele was significantly (p <0.005) higher (OR=1.99 [CI: 1.20-3.33]) than in other patients, there was no difference in major bleeding complications. CONCLUSIONS: Patients bearing CYP2C9 alleles with reduced enzymatic activity have a lower acenocoumarol requirement. In patients with CYP2C9*2 and *3 alleles the frequency of minor bleeding complications and the occurrence of high INR values were significantly higher, but there was no difference in the rate of major bleedings. PMID- 16699990 TI - [Balloon valvuloplasty of critical aortic stenosis performed from an umbilical access in a neonate with endocardial fibroelastosis -- a case report]. AB - A case of percutaneous treatment of neonatal critical aortic stenosis is presented. The baby had endocardial fibroelastosis and borderline dimensions of left heart structures. Umbilical access for the anterograde approach with the use of coronary angioplasty catheter was successfully performed. PMID- 16699989 TI - [No-reflow phenomenon following PCI of venous graft -- a case report]. AB - A case of a 72-year-old male with coronary artery disease, history of two myocardial infarctions, ejection fraction of 30% and CABG performed in 1990, is described. In 2001 he underwent PCI of two grafts. The procedure was complicated by ischaemic stroke. In 2003 the patient underwent another PCI of the occluded venous graft which was complicated by no-reflow phenomenon and acute myocardial infarction. A few months later cardiac resynchronization pacing system was implanted. The patient improved, however, one year later died due to progressive heart failure. Diagnosis and treatment of the no reflow phenomenon are discussed. PMID- 16699991 TI - [Recurrent cardiac arrest due to electro-mechanical dissociation in a patient with variant angina -- a case report]. AB - We present a case of a 44-year-old male with recurrent episodes of cardiac arrest in the course of Prinzmetal's angina. Episodes of variant angina can be life threatening due to episodes of advanced atrioventricular block, asystole, ventricular tachycardia or ventricular fibrillation. It has been suggested to implant an ICD in all patients with variant angina after cardiac arrest. This patient received an ICD, however, he died suddenly 6 months later. The possible mechanism of cardiac arrest was an electromechanical dissociation. PMID- 16699992 TI - [The role of aldosterone in cardiovascular alterations in patients with chronic kidney disease]. PMID- 16699994 TI - [ST segment alterations after ventricular pacing]. PMID- 16699993 TI - [Prevention of venous thromboembolism in older acute hospitalized patients]. PMID- 16699995 TI - [Aneurysms of non-atherosclerotic origin involving right coronary artery and circumflex branch of left coronary artery in a senile patient -- a case report]. PMID- 16699996 TI - [No-reflow phenomenon occurring during elective angioplasty performed due to in stent restenosis -- a case report]. AB - The no-reflow phenomenon is an impairment of microcirculation after successful percutaneous coronary interventions (PCI). The no-reflow phenomenon is usually observed during acute myocardial infarction. This case-report describes no-reflow phenomenon in a patient undergoing elective PCI in the right coronary artery, occluded due to restenosis in implanted stent. After deflation of balloon during angioplasty in restenosed stent, no-reflow phenomenon occurred, followed by asystolia. The patient was successfully resuscitated. During resuscitation procedures, abciximab was administered what improved myocardial perfusion. This case demonstrates that no-reflow phenomenon can be a serious problem during elective PCI, leading even to a cardiac arrest. It shows also the necessity for administration of drugs improving tissue perfusion when no-reflow phenomenon occurs. PMID- 16699997 TI - [Incessant ventricular tachycardia in a patient with a severe aortic stenosis successfully treated by cardiac surgery]. PMID- 16699998 TI - [Cardiosurgery in Poland - 2005]. PMID- 16700001 TI - Changes of sex hormone-binding globulin/SHBG expression in the hypothalamo hypophyseal system of rats during pregnancy, parturition and lactation. AB - Sex hormone-binding globulin (SHBG) is expressed in hypothalamic magnocellular neurons. High co-localization rates of SHBG with oxytocin have been observed in the hypothalamus, indicating that SHBG plays a role in pregnancy, parturition and lactation. Further studies have shown that hypothalamic SHBG expression is malleable to changing steroid conditions. In this study, we have examined SHBG levels in the supraoptic and paraventricular hypothalamic nuclei and in the posterior pituitary lobe of late pregnant, parturient and early lactating rats by IN SITU hybridization, immunocytochemistry, and ELISA. Immunocytochemical and biochemical analysis showed that the SHBG levels increased during late pregnancy in hypothalamic nuclei. During parturition, SHBG levels fell in the magnocellular nuclei but increased in the posterior pituitary lobe. SHBG levels increase again during lactation. At day six of lactation, there was no significant difference in SHBG levels compared to normal cycling female rats, which served as control in this study. IN SITU hybridization showed increased SHBG mRNA signal during late pregnancy. The highest SHBG expression was observed during parturition. Our data indicate that hypothalamic SHBG expression changes during pregnancy, parturition and lactation, parallel to ovarian steroid and co-localized OT levels. This may in part be linked to known steroid actions on synthesis and secretion of magnocellular hypothalamic peptide hormones, important for the control of parturition and lactation. PMID- 16700000 TI - Emerging roles of steroid-binding globulins. PMID- 16700002 TI - Expression of androgen-binding protein (ABP) in human cardiac myocytes. AB - Cardiomyocytes are known to be androgen targets. Changing systemic steroid levels are thought to be linked to various cardiac ailments, including dilated cardiomyopathy (DCM). The mode of action of gonadal steroid hormones on the human heart is unknown to date. In the present study, we used high-resolution immunocytochemistry on semithin sections (1 microm thick), IN SITU hybridization, and mass spectrometry to investigate the expression of androgen-binding protein (ABP) in human myocardial biopsies taken from male patients with DCM. We observed distinct cytoplasmic ABP immunoreactivity in a fraction of the myocytes. IN SITU hybridization with synthetic oligonucleotide probes revealed specific hybridization signals in these cells. A portion of the ABP-positive cells contained immunostaining for androgen receptor. With SELDI TOF mass spectrometry of affinity purified tissue extracts of human myocardium, we confirmed the presence of a 50 kDa protein similar to ABP. Our observations provide evidence of an intrinsic expression of ABP in human heart. ABP may be secreted from myocytes in a paracrine manner perhaps to influence the bioavailabity of gonadal steroids in myocardium. PMID- 16700004 TI - Sex hormone-binding globulin (SHBG) and estradiol cross-talk in breast cancer cells. AB - Sex hormone-binding globulin (SHBG) is a plasma glycoprotein that regulates the action of steroid hormones at several levels. SHBG regulates the availability of free androgens and estradiol to hormone-responsive tissues. Moreover, SHBG is also part of a novel steroid signaling system. We report here on the mechanism of action and the biological effects of SHBG in breast cancer cells, especially distinguishing cross-talk between membrane-initiated SHBG and estradiol pathways. After interacting with a specific binding site on breast cancer cell membranes, SHBG activates a specific pathway, and by cAMP induction, inhibits estradiol mediated activation of ERK. Both estradiol and SHBG membrane-initiated pathways involve cross-talk at MAP kinase level with the ultimate result of inhibiting estradiol-mediated cell growth and antiapoptosis. On the basis of reported evidence, we suggest that SHBG is one of the regulators of growth and apoptosis of estrogen-dependent breast cancer cells. PMID- 16700003 TI - Human sex hormone-binding globulin is expressed in testicular germ cells and not in sertoli cells. AB - The human sex hormone-binding globulin ( SHBG) gene contains at least two transcription units. A 4.3 kb human SHBG transcription unit encodes the precursor polypeptide, which is processed and secreted by hepatocytes as plasma SHBG. The proximal promoter of this transcription unit differs from the corresponding sequence in other mammals, in which it is also expressed in Sertoli cells. In particular, its proximal promoter sequence contains a binding site for USF transcription factors that represses its activity in Sertoli cells. Although human SHBG is not expressed in Sertoli cells, human SHBG transcripts containing an alternative exon 1 sequence are present in testicular germ cells. These are the products of an approximately 8 kb human SHBG transcription unit, and they appear to encode an SHBG isoform that is 4 - 5 kDa smaller than plasma SHBG. This sperm SHBG isoform accumulates between the outer acrosomal membrane and the sperm plasma membrane, and it is released during the capacitation reaction. These remarkable differences in human SHBG expression in the testis, when compared to other mammals, force us to reconsider the functional significance of SHBG expression in the testis in relation to male reproduction. PMID- 16700005 TI - Plasma variation of corticosteroid-binding globulin and sex hormone-binding globulin. AB - Sex hormone-binding globulin (SHBG) and corticosteroid-binding globulin (CBG) circulate in plasma and bind their cognate ligands with high affinity, offering a steroid delivery system to target tissues by a variety of mechanisms. Analysis of these steroid-binding proteins is gaining importance in the clinical setting, although more information is warranted on their diurnal and biological variation. This study shows that plasma SHBG (in normal subjects) exhibits little diurnal or biological variation over the 30 day period studied, in contrast to CBG, where plasma levels peak in the early afternoon. This leads to attenuation of the diurnal free cortisol level rhythm compared to total cortisol. We also show that plasma CBG is significantly lower in male subjects with the metabolic syndrome compared to age-matched lean counterparts, and may therefore act as a surrogate marker of insulin resistance. The consequence of lower levels of CBG in these obese male subjects is reflected by higher levels of circulating free cortisol, potentially offering a more favourable environment for adipogenesis. PMID- 16700007 TI - Expression of corticosteroid-binding protein in the human hypothalamus, co localization with oxytocin and vasopressin. AB - Corticosteroid-binding globulin, a specific steroid carrier in serum with high binding affinity for glucocorticoids, is expressed in various tissues. In the present study, we describe the immunocytochemical distribution of this protein in neurons and nerve fibers in the human hypothalamus. CBG immunoreactive perikarya and fibers were observed in the paraventricular, supraoptic, and sexual dimorphic nuclei in the perifornical region, as well as in the lateral hypothalamic and medial preoptic areas, the region of the diagonal band, suprachiasmatic and ventromedial nuclei, bed nucleus of the stria terminalis and some epithelial cells from the choroid plexus and ependymal cells. Stained fibers occurred in the median eminence and infundibulum. Double immunostaining revealed a partial co localization of corticosteroid-binding globulin with oxytocin and, to a lesser extent, with vasopressin in the paraventricular and the supraoptic nuclei. Double immunofluorescence staining showed coexistence of these substances in axonal varicosities in the median eminence. We conclude that neurons of the human hypothalamus are capable of expressing corticosteroid-binding globulin, in part co-localized with the classical neurohypophyseal hormones. The distribution of CBG immunoreactive neurons, which is widespread but limited to specific nuclei, indicates that CBG has many physiological functions that may include neuroendocrine regulation and stress response. PMID- 16700006 TI - Expression of corticosterone-binding globulin in the rat hypothalamus. AB - We observed coexistence of corticosteroid-binding globulin (CBG) with vasopressin (VP) and oxytocin (OT) in magnocellular neurons in rat hypothalamus by combined immunoperoxidase staining and immunofluorescence. A portion of the supraoptic and of the paraventricular neurons showed double immunostaining of CBG with either VP or with OT. CBG staining was intensified by pretreating animals with colchicine to block axonal transport. CBG was also observed in widespread axonal projections throughout the lateral hypothalamus, the median eminence and the posterior pituitary lobe. Single ependymal cells and some of the endocrine cells in the anterior lobe contained specific CBG immunoreactivity. IN SITU hybridization of semithin sections with a synthetic oligonucleotide probe to CBG mRNA provided staining of magnocellular hypothalamic neurons, but not ependymal cells or anterior lobe cells. Western blots of CBG extracted by affinity chromatography from hypothalamus homogenates showed a band at approximately 50 kDa. Our observations indicate the intrinsic expression of CBG in peptidergic hypothalamus neurons in rat. The multiple locations of CBG-expressing neurons indicate multiple functional properties, probably exceeding the role of a mere steroid transporter. CBG is likely to be subject to axonal transport and secretion in a neuropeptide-like fashion, perhaps involved in neuroendocrine regulation, which may include stress responses. PMID- 16700008 TI - Plasma-binding globulins and acute stress response. AB - Within studies of acute stress physiology an increase in glucocorticoid secretion is thought to be the primary mediator of tissue response to stress. Corticosteroid-binding globulin may regulate tissue availability of steroids, but has not been considered a dynamic component of the acute stress response. Here, we examined CBG level over the common 60-minute time frame in an acute capture and handling protocol to investigate whether CBG capacity is dynamic or static over short stressors. Using a comparative approach, we measured CBG response to capture and handling stress in nine species of birds, representing five orders and nine families. CBG capacity significantly declined within 30 - 60 minutes of capture in five of the nine species examined. This decline may serve to significantly increase the level of corticosterone reaching tissues during acute stress. PMID- 16700009 TI - The retinol-binding protein system: a potential paradigm for steroid-binding globulins? AB - Retinol (vitamin A) is an example of a small molecule that is essential for higher organisms; its utilisation has been involved in the evolution of a number of proteins. In mammalian species, retinol is obtained from the diet and controls the release of its binding protein from hepatocytes into the blood stream. Subsequent influx into cells under normal situations usually involves a specific membrane-bound receptor for retinol-binding protein, which facilitates the uptake of retinol alone or bound to its carrier. This specific receptor has not yet been identified, but a receptor for a related lipocalin has been cloned. It represents a relatively new family, and there are a number of related genes in various eukaryotic genomes, suggesting that the system is very widespread in multicellular organisms. Its significance has been highlighted recently by the suggestion that retinol-binding protein, through its receptor, may play a major role in type 2 diabetes, perhaps the greatest scourge of modern society. This system may provide a new paradigm in mammalian biology, another example of which may exist in the processes responsible for steroid handling. This review outlines the characteristics of retinol utilisation in mammalian species, focusing primarily on the uptake system. PMID- 16700010 TI - The role of plasma-binding proteins in the cellular uptake of lipophilic vitamins and steroids. AB - Steroid hormones and many other lipophilic compounds are believed to enter cells solely by free diffusion through the plasma membrane. However, recent work using a megalin-deficient mouse model has identified a new endocytic pathway responsible for the delivery of steroids to renal and gonadal tissues. This review describes these new pathways for uptake of 25-hydroxy-vitamin-D3 and the gonadal sex-steroids (17beta-estradiol and testosterone) bound to vitamin D binding protein and sex hormone-binding globulin respectively. Furthermore examples of other lipophilic molecules that enter cells by receptor-mediated pathways will be presented and the receptors responsible for their uptake described. PMID- 16700011 TI - Co-transport of sex hormone-binding globulin/SHBG with oxytocin in transport vesicles of the hypothalamo-hypophyseal system. AB - The intrinsic expression of sex hormone binding globulin (SHBG) in magnocellular hypothalamic neurons, in part co-localized with either vasopressin or oxytocin, was recently described. This study is focused on the ultrastructural localization of SHBG in the hypothalamo-neurohypophyseal pathway in rats. Immunostaining for SHBG in the hypothalamic perikarya was increased by colchicine treatment, indicating that the steroid-binding globulin is subject to rapid axoplasmic transport along with the classical posterior lobe peptides. With immunoelectron microscopic double labeling, we found co-localization of oxytocin and sex hormone binding globulin in a portion of the large dense-core vesicles in paraventricular and supraoptic perikarya and in axonal varicosities in the median eminence and in the posterior lobe. Our observations show that SHBG is processed, transported and stored along with oxytocin suggesting that SHBG is released from nerve terminals in the posterior lobe, the median eminence and possibly the brain similarly to and in conjunction with oxytocin. PMID- 16700012 TI - Recent advances in antimalarial drug development. AB - Malaria caused by protozoa of the genus Plasmodium, because of its prevalence, virulence, and drug resistance, is the most serious and widespread parasitic disease encountered by mankind. The inadequate armory of drugs in widespread use for the treatment of malaria, development of strains resistant to commonly used drugs such as chloroquine, and the lack of affordable new drugs are the limiting factors in the fight against malaria. These factors underscore the continuing need of research for new classes of antimalarial agents, and a re-examination of the existing antimalarial drugs that may be effective against resistant strains. This review provides an in-depth look at the most significant progress made during the past 10 years in antimalarial drug development. PMID- 16700013 TI - Diffusion-weighted magnetic resonance imaging in the syndrome of acute bilateral basal ganglia lesions in diabetic uremia. AB - In this report, we have presented a diabetic patient with uremia, in which acute Parkinsonism occurred, coupled with acute mental confusion, after a sudden increase in blood urea nitrogen and serum creatinin levels. Diffusion-weighted magnetic resonance imaging revealed a unique cytotoxic-type edema in the bilateral basal ganglia during the acute phase. Signal alterations were shown to regress in accordance with the normalized apparent diffusion coefficient (ADC) values, but irreversible cystic degeneration developed in the globus pallidus, with the very low preceding ADC values. PMID- 16700016 TI - Assessment for crisis intervention. AB - This article describes the triage assessment system (TAS) for crisis intervention. The TAS assesses affective, behavioral, and cognitive reactions of individuals to crisis events. This assessment model offers clinicians an understanding of the type of reactions clients are experiencing as well as the intensity of these reactions. The TAS provides a quick, accurate, and easy-to- use method that is directly usable in the intervention process. The system can also be used to monitor clients' progress during the intervention process. Two case illustrations are presented to demonstrate the use of the model. In addition, the Triage Assessment Form: Crisis Intervention is included as an Appendix. PMID- 16700014 TI - Redox-sensitive contrast agents for MRI based on reversible binding of thiols to serum albumin. AB - DOTA-based complexes of gadolinium (Gd) bearing a thiol moiety on a propyl or hexyl arm were synthesized. It was hypothesized that these complexes would form reversible covalent linkages with human serum albumin (HSA), which contains a reactive thiol at cysteine-34. The binding constant of the hexyl complex to HSA was measured to be 64 mM(-1) and decreased to 17, 6.1, and 3.6 mM(-1) in the presence of 0.5, 1, and 2 mM homocysteine, respectively. The binding constant of the propyl complex to HSA was significantly lower (5.0 mM(-1)) and decreased to 2.0, 1.5, and 0.87 mM(-1) in the presence of 0.5, 1, and 2 mM homocysteine, respectively. The longitudinal water-proton relaxivities of the hexyl and propyl complexes at 37 degrees C and 4.7 T were 2.3 and 2.9 mM(-1) s(-1), respectively, in saline. The relaxivities of the HSA-bound forms of the hexyl and propyl complexes were calculated to be 5.3 and 4.5 mM(-1) s(-1), respectively. The in vivo pharmacokinetics of both thiol complexes were altered by a chase of homocysteine but not saline, while the washout of GdDTPA was unaffected by either chase. Such redox-sensitive reversible binding of Gd complexes to plasma albumin can be exploited for imaging tissue redox and the blood-pool by MRI. PMID- 16700017 TI - Resilience in the face of potential trauma: clinical practices and illustrations. AB - Many persons exposed to loss or potentially traumatic events manage the stresses of these experiences with minimal to no impact on their daily functioning. The prevalence of this resilient capacity has surprised researchers and clinicians alike and refocused clinical practice. We review three key points about resilience: resilience is different from the process of recovery; resilience in the face of loss or potential trauma is common; and there are multiple and sometimes unexpected pathways to resilience. We then present six clinical practices informed by the study of resilience, illustrating key points with clinical vignettes. PMID- 16700018 TI - Noways tired: perspectives of clinicians of color on culturally competent crisis intervention. AB - The authors demonstrate through case material the clinical benefits of utilizing a culturally competent approach to crisis intervention. The focus here is on intervention with racial/ethnic minorities, in particular Black Americans, Latino Americans and Asian Americans; but the authors also address the importance of culturally competent crisis intervention praxis for all clients. PMID- 16700020 TI - Psychotherapy for families in the aftermath of a disaster. AB - We have not seen the last of natural disasters; nor have we endured the last of the terrorist acts. This article offers clinical guidance in providing mental health services to families in the aftermath of such events. The trauma experienced by an individual victimized by a natural disaster or terror attack reverberates through an entire family. Although based in established clinical practice, intervention with survivors of disasters is a different process from other treatments. A case illustrates the key principles and central methods of family therapy with a traumatized family. PMID- 16700019 TI - The aftermath of disaster: children in crisis. AB - This article uses examples from the terrorist attacks on the World Trade Center and the hurricane disasters of Katrina and Rita to illustrate the impact of crisis in the lives of children and adolescents. The author reviews children's responses to loss and crisis. Therapeutic approaches that facilitate integration of crisis and loss are provided, including illustrations of self-care, comfort strategies, and developmental, traditional, and nontraditional methods. PMID- 16700021 TI - Community mental health response to crisis. AB - Mental health is seen as an important aspect in the community response to crisis and disaster. Research offers limited guidance to what interventions are likely to be effective in preventing postdisaster mental health problems. This article reviews recommended elements of mental health responses to community disasters. Different factors influencing response are illustrated by using 9/11 and Hurricane Katrina as examples. Clinical suggestions for community planning and actions of individual psychologists conclude the article. PMID- 16700022 TI - Cognitive behavioral therapy for prolonged postdisaster distress. AB - In this article, we present and illustrate cognitive-behavioral therapy for postdisaster distress. The treatment is for individuals who show more than normal transient stress after disaster and functions as an intermediate step between traditional crisis counseling and longer-term mental health treatments. Thus, it is one part of a larger mental health disaster response and is designed to be implemented no sooner than 60 days post disaster. A clinical case demonstrates its process and technique in a community practice setting. PMID- 16700023 TI - Source of nonlinearity in echo-time-dependent BOLD fMRI. AB - Stimulation-induced changes in transverse relaxation rates can provide important insight into underlying physiological changes in blood oxygenation level dependent (BOLD) contrast. It is often assumed that BOLD fractional signal change (DeltaS/S) is linearly dependent on echo time (TE). This relationship was evaluated at 9.4 T during visual stimulation in cats with gradient-echo (GE) and spin-echo (SE) echo-planar imaging (EPI). The TE dependence of GE DeltaS/S is close to linear in both the parenchyma and large vessel area at the cortical surface for TEs of 6-20 ms. However, this dependence is nonlinear for SE studies in the TE range of 16-70 ms unless a diffusion-weighting of b = 200 s/mm(2) is applied. This behavior is not caused by inflow effects, T(2)* decay during data acquisition in SE-EPI, or extravascular spin density changes. Our results are explained by a two-compartment model in which the extravascular contribution to DeltaS/S vs. TE is linear, while the intravascular contribution can be nonlinear depending on the magnetic field strength and TE. At 9.4 T, the large-vessel IV signal can be minimized by using long TE and/or moderate diffusion weighting. Thus, stimulation-induced relaxation rate changes should be carefully determined, and their physiological meanings should be interpreted with caution. PMID- 16700024 TI - Phase-contrast velocimetry with hyperpolarized 3He for in vitro and in vivo characterization of airflow. AB - This paper describes a technique that combines radial MRI and phase contrast (PC) to map the velocities of hyperpolarized gases ((3)He) in respiratory airways. The method was evaluated on well known geometries (straight and U-shaped pipes) before it was applied in vivo. Dynamic 2D maps of the three velocity components were obtained from a 10-mm slice with an in-plane spatial resolution of 1.6 mm within 1 s. Integration of the in vitro through-plane velocity over the slice matched the input flow within a relative precision of 6.4%. As expected for the given Reynolds number, a parabolic velocity profile was obtained in the straight pipe. In the U-shaped pipe the three velocity components were measured and compared to a fluid-dynamics simulation so the precision was evaluated as fine as 0.025 m s(-1). The technique also demonstrated its ability to visualize vortices and localize characteristic points, such as the maximum velocity and vortex center positions. Finally, in vivo feasibility was demonstrated in the human trachea during inhalation. PMID- 16700025 TI - Velocity-selective arterial spin labeling. AB - In pathologies in which slow or collateral flow conditions may exist, conventional arterial spin labeling (ASL) methods that apply magnetic tags based on the location of arterial spins may not provide robust measures of cerebral blood flow (CBF), as the transit delay for the delivery of blood to target tissues may far exceed the relaxation time of the tag. Here we describe current methods for ASL with velocity-selective (VS) tags (termed VSASL) that do not require spatial selectivity and can thus provide quantitative measures of CBF under slow and collateral flow conditions. The implementation of a robust multislice VSASL technique is described in detail, and data obtained with this technique are compared with those obtained with conventional pulsed ASL (PASL). The technical considerations described here include the design of VS pulses, background suppression, anisotropy with respect to velocity-encoding directions, and CBF quantitation issues. PMID- 16700026 TI - Magnetic field correlation imaging. AB - A magnetic resonance imaging (MRI) method is presented for estimating the magnetic field correlation (MFC) associated with magnetic field inhomogeneities (MFIs) within biological tissues. The method utilizes asymmetric spin echoes and is based on a detailed theory for the effect of MFIs on nuclear magnetic resonance (NMR) signal decay. The validity of the method is supported with results from phantom experiments at 1.5 and 3 T, and human brain images obtained at 3 T are shown to demonstrate the method's feasibility. The preliminary results suggest that MFC imaging may be useful for the quantitative assessment of iron within the brain. PMID- 16700027 TI - PINK1 homozygous W437X mutation in a patient with apparent dominant transmission of parkinsonism. AB - We analyzed the PINK1 gene in 58 patients with early-onset Parkinsonism and detected the homozygous mutation W437X in 1 patient. The clinical phenotype was characterized by early onset (22 years of age), good response to levodopa, early fluctuations and dyskinesias, and psychiatric symptoms. The mother, heterozygote for W437X mutation, was affected by Parkinson's disease and 3 further relatives were reported affected, according to an autosomal dominant transmission. PMID- 16700028 TI - Hyaluronan and its receptor CD44 in the heart of newborn and adult rats. AB - The extracellular matrix is a dynamic space and a prerequisite for the function of cardiomyocytes. We have previously reported on the distribution of the glycosaminoglycan hyaluronan (HYA) and its cellular receptor CD44 in the vascular system. In newborn rats, HYA and its receptor were colocalized, but in the adult animals, no such colocalization was observed. Furthermore, the distribution of both HYA and CD44 differed between newborn and adult animals. In this study, the distribution of HYA and its receptor CD44 is explored in the heart. Hearts from newborn and adult rats were stained for visualization of HYA and CD44 using histochemistry and immunohistochemistry. HYA stained the interstitium of the myocardium heterogeneously. Strong staining was seen in the heart valves of both newborn and adult animals. CD44 staining was sparse in hearts from both newborn and adult animals. There are no major changes in the distribution of HYA in the myocardium during the postnatal development in contrast to the blood vessels. Thus, the structure of the interstitium does not change after birth when the heart starts to grow mainly through cardiomyocyte hypertrophy rather than hyperplasia. The abundance of HYA in the heart valves is probably related to their unique physiological properties to withstand repetitive mechanical stress. PMID- 16700029 TI - Impaired rhythm generation in essential tremor. AB - It has been suggested that the cerebellum plays a role in the event-based timing of synchronized repetitive movements. We hypothesized that regularity of rhythmic movements in essential tremor (ET) is impaired, since several lines of evidence suggest the involvement of the cerebellum in the pathomechanism of ET. To test this assumption, we examined the regularity and the maximum frequency of auditory paced repetitive movements at slow and fast stimulus rate in 34 ET patients. Variability of rhythmic finger tapping and alternating hand movements, defined by the standard deviation of movement offset before or after the pacing signal, was significantly higher compared to healthy controls. Timing of rhythmic movements of the two hands was disturbed to the same degree. Our results suggest a severe deficit of event-based rhythm generation on both sides in ET, supporting the presumed bilateral cerebellar dysfunction in this disorder. PMID- 16700030 TI - Hemiballism-Hemichorea in presentation of cranial tuberculoma. PMID- 16700031 TI - Online movement control in multiple sclerosis patients with tremor: effects of tendon vibration. AB - Patients with intention tremor due to multiple sclerosis (MS) exhibit an increased reliance on visual feedback in the sensorimotor control of slow goal directed movements. In the present study, the use of proprioceptive information was investigated in MS patients with intention tremor compared to MS patients without tremor and healthy controls. Tendon vibration was applied to the wrist extensor muscles during a memory-guided slow wrist step-tracking task to investigate the use of muscle spindle afferent information in online movement control. A significant reduction of movement amplitude was induced by tendon vibration in all three groups, but the effect was found to be smaller in MS patients with tremor (28%) than in subjects without tremor (50%). Vibration also induced an increase of overall tremor amplitude in the MS tremor group; however, its effect on movement amplitude was not directly related to (changes in) tremor severity. The results suggest that the decreased online use of proprioceptive information in MS patients with tremor reflects an adaptation over time to cope with a tremor-related noisy background. Abnormalities in proprioceptive processing may explain why MS patients with tremor show an increased reliance on visual feedback for online motor control. PMID- 16700032 TI - Assessment of simple movements reflects impairment in Huntington's disease. AB - Clinical rating, caudate atrophy, disturbed movement performance, neuropsychological testing, and age-related genetic disease load (CAG index) are tools that reflect impairment after onset of Huntington's disease (HD). Objectives were to compare scored HD symptoms, results of neuropsychological testing and of instrumental measurement of simple motion sequences, assess caudate atrophy and CAG index, and investigate their relation to each other in 131 subjects of various HD stages. Caudate atrophy and CAG index significantly increased in advanced HD patients. Motor test results significantly differed between HD patients and 49 controls, but not between HD gene carriers and controls. Instrumental test outcomes, scored HD intensity, caudate atrophy, and CAG index significantly correlated to each other. Neuropsychological testing, which we only performed in the HD gene carriers and the previously untreated HD patients, reflected the early appearance of HD symptoms and correlated with the motor test results. Results of our applied instrumental tool measure impaired movement performance, which is not specific for HD, but reflects the various methods assessed and the slowly evolving symptoms of the degenerative process in HD. PMID- 16700033 TI - Parkinsonism and personality changes following an acute hypoxic insult during mountaineering. PMID- 16700034 TI - Outcomes in patients with splenic marginal zone lymphoma and marginal zone lymphoma treated with rituximab with or without chemotherapy or chemotherapy alone. AB - BACKGROUND: The optimal management of patients with splenic marginal zone lymphoma/marginal zone lymphoma (SMZL) is controversial. The objective of this retrospective study was to compare the outcomes of patients with SMZL who received treatment with rituximab, rituximab plus chemotherapy, or chemotherapy alone. METHODS: The Leukemia Service database was searched for patients with splenic lymphoma who were registered between May 1995 and October 2004. The indications for treatment were the same as those used for patients with chronic lymphocytic leukemia. RESULTS: SMZL was confirmed in 70 patients. The median age was 64 years. The median number of CD20 molecules per cell was 69 x 10(3). Forty three patients required systemic therapy; rituximab in 26 patients, chemotherapy plus rituximab in 6 patients, and chemotherapy alone in 11 patients. Ten additional patients underwent splenectomy, and 17 patients were in the observation group. The overall response rates were 88% with rituximab, 83% with rituximab plus chemotherapy, and 55% with chemotherapy alone; the 3-year survival rates were 95%, 100%, and 55%, respectively. The 3-year failure-free survival (FFS) rates were 86%, 100%, and 45% in the rituximab, rituximab plus chemotherapy, and chemotherapy alone groups, respectively. Rituximab treatments resulted in longer survival and FFS compared with chemotherapy. Rituximab alone resulted in disappearance of splenomegaly in 92% of patients and normalization of absolute lymphocyte counts. In univariate analysis, younger age and rituximab based therapy were predictive of longer FFS. CONCLUSIONS: Rituximab with or without chemotherapy was found to have major activity in patients with SMZL. These results may be associated with high levels of cellular CD20 antigen sites. Rituximab should be the treatment of choice, at least in older patients with SMZL who have comorbid diseases. PMID- 16700035 TI - Treatment options in lymph node-positive prostate cancer. AB - With improved awareness and screening, the incidence of lymph node-positive prostate cancer has declined dramatically over the last 50 years. Stage of cancer, prostate-specific antigen, and grade are risk factors for positive lymph nodes; and those factors, along with the number of involved lymph nodes, are prognostic factors for outcome. Although the numbers have declined, the number of men with lymph node-positive prostate cancer remains significant, and the current challenge is how best to treat these patients. Commonly used treatments include any combination of androgen ablation, surgery, and radiation. There have been a few studies with chemotherapy, and no treatment has been proven superior to the others. Consequently, there remain several reasonable alternatives to treatment, and long-term survival is not unusual. PMID- 16700036 TI - Central nervous system recurrence in adult patients with acute lymphoblastic leukemia: frequency and prognosis in 467 patients without cranial irradiation for prophylaxis. AB - Recurrence of acute lymphoblastic leukemia (ALL) in the central nervous system (CNS) confers a poor prognosis, although to the authors' knowledge, only a few studies have analyzed this issue in adults. For the current study, the authors analyzed the frequency, predictive factors, and prognosis of CNS involvement and recurrence in adult patients with ALL who did not receive cranial irradiation for CNS prophylaxis. Four hundred sixty-seven adult patients (age > or = 15 years) with ALL were treated on 4 protocols: ALL-89 (standard-risk and high-risk ALL; n = 108 patients), ALL-93 (high-risk ALL; n = 222 patients), ALL-96 (standard-risk ALL; n = 84 patients), and ALL3-97 (Burkitt leukemia; n = 53 patients). CNS prophylaxis consisted of intrathecal methotrexate, cytarabine, and hydrocortisone together with high-dose systemic methotrexate and cytarabine. The mean age (+/- standard deviation) was 33 years (+/- 16 years), and 272 patients were males. ALL subtypes included an early pre-B phenotype (15%), a common phenotype (45%), a pre B phenotype (5%), a mature B phenotype (11%), and a T phenotype (24%). CNS involvement at diagnosis was observed in 18 patients (3.9%). Of 159 recurrences, 22 occurred (5.8%) in the CNS (14 isolated and 8 combined). A lactate dehydrogenase level > 1000 U/L was the only factor associated with the risk of CNS recurrence. A complete remission was attained in 7 of 22 patients (32%). The median overall survival after recurrence was 0.7 years for patients with isolated CNS recurrence, 0.13 years for patients with combined recurrence, and 0.41 years for patients with bone marrow recurrence (P = .11). The only 2 survivors underwent stem cell transplantation. The frequency of CNS recurrence in adult patients with ALL who do not receive radiotherapy for CNS prophylaxis was similar to the frequency observed in protocols that included cranial irradiation. A lactate dehydrogenase value >1000 U/L was the only factor found to be associated with CNS recurrence. The prognosis for patients who develop CNS recurrence is poor, identical to that for patients who develop bone marrow recurrence. PMID- 16700037 TI - Radical prostatectomy for clinical T4 prostate cancer. AB - BACKGROUND: Occasionally, patients with clinical T4 (cT4) prostate cancer undergo surgery. Published data on outcomes after radical prostatectomy (RP) in patients with such advanced stage disease and on the impact of adjuvant radiation therapy (RT) and hormone therapy (HT) are nonexistent. METHODS: Data from the Surveillance Epidemiology and End Results (SEER) data base were reviewed for the 7-year period from 1995 to 2001. Specifically, data were analyzed for 1093 patients with cT4, lymph node-negative or lymph node-positive, M0 prostate cancer without distant lymph node involvement or a history of other cancer. Using follow up data through 2002, postdiagnosis survival was examined in 5 treatment groups: radical prostatectomy (RP) either alone or in combination with other therapy, radiation therapy (RT) alone, hormone therapy (HT) alone, RT plus HT, and no treatment (NT). All results were expressed as 1-year, 3-year, and 5-year observed survival and corresponding relative survival. Mortality across treatment categories was compared by using a Cox proportionate hazards model controlling for age, year of diagnosis, race, tumor grade, regional lymph node involvement, clinical tumor extension, and SEER registry. RESULTS: Observed and relative survival rates were lowest among patients who received NT and highest among patients who underwent RP. Adding adjuvant RT or HT to RP conferred no survival benefit. Multivariate survival analyses revealed a significant increase in mortality among HT-only patients and among patients who received NT compared with patients who underwent RP. The differences in survival among treatment types were most pronounced in a relatively small group of patients who had positive regional lymph node extension. In all other patients, the results suggested a modest (but not significant) improvement in survival after RT plus HT. CONCLUSIONS: SEER data revealed that patients who underwent RP for cT4 prostate cancer had increased survival compared with patients who received RT alone or HT alone and had a survival comparable to that of patients who received RT plus HT. The benefit of RP appears to be limited to a relatively small subset of patients who have regional lymph node extension. PMID- 16700038 TI - Expression of estrogen receptors-alpha and -beta in bladder cancer cell lines and human bladder tumor tissue. AB - BACKGROUND: Estrogen receptors (ERs) are known to mediate important physiologic responses as well as the growth of some tumors in response to estradiol stimulation. In a previous study the selective ER modulator raloxifene was shown to induce apoptosis in an ERbeta-positive bladder cancer cell line. However, the expression of ERbeta in human bladder cancer has not been thoroughly investigated. METHODS: ERalpha and ERbeta expression in 224 bladder tumor samples was evaluated using tissue microarray and immunohistochemistry. Levels of ERalpha and ERbeta protein and mRNA expression were determined in several bladder cancer cell lines using quantitative reverse-transcriptase polymerase chain reaction (RT PCR) and Western blot analysis. The effect of estradiol and antiestrogen treatments on RT4 bladder cancer cell growth was determined by cell proliferation assays. RESULTS: Analyses revealed that only 2 human bladder cancers weakly expressed ERalpha. In contrast, the expression of ERbeta was detected in 141 tumors (63%). ERbeta was expressed in 58% of WHO Grade 1 and 2 tumors, whereas 70% of Grade 3 tumors demonstrated expression (P = .085). Importantly, although only 53% and 55% of Ta and T1 tumors demonstrated ERbeta expression, 80% of T2, 81% of T3, and 75% of T4 tumors showed ERbeta expression. The differences in ERbeta expression between Ta/T1 and T2/T3/T4 tumors were found to be highly significant (P < .001). Metastatic transitional cell carcinomas had ERbeta expression (80%) comparable to that of muscle invasive bladder cancers. Western blot analysis detected ERbeta protein expression in each of the 5 bladder cancer cell lines tested, whereas no or very low levels of ERalpha were found. Quantitative RT-PCR revealed that higher levels of ERbeta than ERalpha mRNA were present in 5637, T-24, TSU-Pr1, and TCC-Sup bladder cancer cells, whereas ER alpha mRNA levels were greater than ERbeta in RT4 cells. Treatment with 17beta estradiol modestly increased RT4 cell growth, whereas the antiestrogens, 4 hydroxtamoxifen, raloxifene, or ICI 182,780 inhibited the growth of RT4 cells. CONCLUSIONS: ERbeta is the dominant receptor expressed in bladder cancer cell lines and in the majority of human bladder tumors. Moreover, the degree of ERbeta expression increases with increasing stage and grade of differentiation. Antiestrogens have an inhibitory effect on the growth of bladder cancer cells in vitro. PMID- 16700039 TI - Primary bone lymphoma: treatment results and prognostic factors with long-term follow-up of 82 patients. AB - BACKGROUND: To the authors' knowledge, there is limited information on the preferred treatment and long-term prognosis of primary bone lymphomas (PBLs). All PBL cases treated at the study center between 1963 and 2003 were analyzed to determine patient, disease, and treatment factors that could affect outcome measured by overall survival (OS), cause-specific survival (CSS), and freedom from-treatment failure (FFTF). METHODS: A total of 101 patients with PBL diagnosed at the study institution were identified. Nineteen patients were excluded because they transferred their treatment or follow-up to another center. Disease control, survival, and prognostic factors were analyzed for all 82 remaining patients. RESULTS: The median age of the patients was 48 years (range, 11-83 years). Approximately 80% presented with diffuse large-cell lymphoma (DLCL), and 81% presented with Ann Arbor Stage I or II disease. Approximately 57% were treated with combined modality therapy, 14% were treated with radiation therapy alone, and 30% were treated with chemotherapy alone. The median follow-up was 67 months (range, 2-280 months). The 5-year OS, CSS, and FFTF were 88%, 96%, and 81%, respectively. The 5-year OS for patients treated with combined modality versus single-modality therapy was 95% versus 78% (P = .013), and the 5-year FFTF for patients treated with combined modality versus single-modality therapy was 90% versus 67% (P = .025). The 5-year CSS for patients treated with combined modality versus single-modality therapy was 95% versus 83% (P = .065). Using a Cox regression for multivariate analysis, age < 40 years and use of combined modality therapy were found to be favorable prognostic factors for OS, CSS, and FFTF. CONCLUSIONS: To the authors' knowledge, the current study is the largest series of patients with PBL treated with modern curative modalities. The data demonstrate that primary lymphoma involving the bone has an excellent prognosis. Patients with PBL treated with combined modality versus single modality therapy were found to have a superior outcome, with a significantly better survival. PMID- 16700040 TI - Cerebral metastases pathology after radiosurgery: a multicenter study. AB - BACKGROUND: To the authors' knowledge, comprehensive human pathologic investigations that explore fundamental radiosurgical effects on metastatic brain tumors are sparse in the literature. The objective of this study was to analyze histopathologic findings in a set of clinically recurrent cerebral metastases after patients underwent stereotactic radiosurgery (SRS). METHODS: In a series of 7500 patients who underwent radiosurgery, 2020 patients (27%) harbored cerebral metastases. Eighteen of 2020 patients (0.9%) underwent subsequent craniotomy for tumor removal anywhere from 1 month to 59 months after they received high-dose irradiation. Histologic and immunohistochemical investigations were performed on the surgically resected tissue specimens. These specimens were within the radiosurgical treatment volume of the metastatic tumor. RESULTS: Light microscopy revealed 3 basic categories of histologic responses: acute-type, subacute-type, and chronic-type tissue reactions. A moderate-to-intense inflammatory cell reaction was seen in the tissue responses of well controlled neoplasms (i.e., in patients who had neoplasms that required craniotomy for recurrent disease > 5 months after SRS), whereas the inflammatory reaction was missing or sparse in poorly controlled neoplasms (patients who required craniotomy for recurrent disease < 5 months after SRS). This reaction was seen within the irradiated tumor volume and not in the peritumoral area nor in areas remote from the radiosurgical treatment volume. Immunohistochemical characterization demonstrated the presence of prominent CD68-positive macrophage and CD3-positive T-lymphocyte populations. A progressively severe vasculopathy also was observed with increasing time after radiosurgery. CONCLUSIONS: Although causality has not been established, a brisk inflammatory response and more severe vasculopathy were observed in lesions in which recurrences were more delayed. PMID- 16700041 TI - Bone invasion in extremity soft-tissue sarcoma: impact on disease outcomes. AB - BACKGROUND: The purpose of the current study was to evaluate histologic bone invasion as a predictor of oncologic outcome in extremity soft-tissue sarcoma (STS) patients presenting to a specialty sarcoma center between 1986 and 2001. METHODS: All patients who underwent surgery for extremity STS were identified from the prospective database at the study institution. Patient demographic features were compared using chi-square analyses or independent-sample Student t tests. The disease outcomes were compared for those with and without bone invasion using Kaplan-Meier survival analysis and Cox modeling. RESULTS: In a review of 874 patients with STS of the extremity, 48 patients (5.5%) had evidence of bone invasion. Patients with bone invasion presented with larger tumors that were more frequently deep in the extremity and more often had metastases at presentation. Patients with bone invasion had lower metastasis-free survival but bone invasion was not found to be an independent prognostic factor (P = .274) on Cox modeling. Bone invasion was found to be prognostic of overall survival on multivariate analysis (P < .0001). CONCLUSIONS: In a small percentage of patients with STS, bone resection may be required to obtain an adequate surgical margin, thereby limiting the risk of local tumor recurrence. Histologic bone invasion portends a poorer prognosis in terms of overall survival. PMID- 16700043 TI - Mortality experiences among 15+ year survivors of childhood and adolescence cancers. PMID- 16700044 TI - Life threatening parvovirus B19 and herpes simplex virus associated acute myocardial dysfunction in a child with homozygous sickle cell disease. AB - Human parvovirus (HPV) B19, a common infection, frequently causes transient red cell aplasia in children with hemolytic anemia, such as sickle cell disease (SCD). It was considered to be a self-limited condition, easily treated with blood transfusion. However, acute splenic sequestration, acute chest syndrome, nephrotic syndrome, and stroke have been reported in SCD patients following HPV B19 infection. We report a 3-year-old child with SCD who developed fulminant myocarditis following HPV B19-related aplastic crisis. The diagnosis of myocarditis should be considered in a patient with hemolytic anemia with an infection with HPV B19 who develops signs of cardiopulmonary failure despite correction of anemia. PMID- 16700046 TI - Excess burden of constipation in Parkinson's disease: a pilot study. AB - An analysis was undertaken of clinic-based questionnaires that asked people with Parkinson's disease and a control group of older people without a known neurological condition about their experiences of constipation. People with Parkinson's disease report higher constipation on a validated objective measure, the Rome criterion (59% vs. 20.9%); a behavioral indicator, laxative-taking (38.4% vs. 14.2%); and subjective self-report of being always or often concerned by it (33.4% vs. 6.1%). Many people with Parkinson's disease experience constipation problems but they may not bring these to the attention of their healthcare providers. More research is required to understand the causes and management options. PMID- 16700047 TI - Age distributions, birth weights, nephrogenic rests, and heterogeneity in the pathogenesis of Wilms tumor. AB - BACKGROUND: The National Wilms Tumor Study (NWTS) constitutes a unique resource for study of clinical, pathologic, and epidemiologic features of Wilms tumor (WT). PROCEDURE: Data from NWTS-3,4,5 were compiled for 7,455 patients with tumors of favorable (FH) or anaplastic (AH) histology. The associations of birth weight (BW) and age-at-onset with gender, intralobar (ILNR), and perilobar (PLNR) nephrogenic rests, tumor focality, congenital malformation syndromes, and tumor histology were analyzed using descriptive statistics and linear regression. RESULTS: Mean BWs for male and female patients without PLNR were 3.52 and 3.36 kg, respectively, and for those with PLNR were 0.12 kg and 0.15 kg heavier. Mean age was 45 months for males with no rests whose tumors were unifocal and of triphasic favorable histology. ILNR or multifocality decreased the mean age by 18 and 10 months, respectively, whereas female gender, blastemal/FH or AH increased it by 3, 10, and 16 months. Over 90% of multifocal tumors occurred in the presence of demonstrated ILNR or PLNR or both. The apparent bimodality of the age distributions and later mean ages-at-onset for females with both unifocal and multifocal tumors were explained in part by the relative deficit in females of ILNR versus PLNR-associated tumors. CONCLUSIONS: These observations support the view that there are multiple pathways to Wilms tumorigenesis. They will facilitate selection of informative subgroups of patients for molecular analysis that may serve to identify the putative pathway for the majority of patients who cannot be classified provisionally on the basis of ILNR or PLNR. PMID- 16700048 TI - Bilateral germinoma of the basal ganglia. AB - Germinoma arising in the bilateral basal ganglia is exceedingly rare, with only five cases reported to date. Owing to non-specific clinical findings and the frequent presence of ill-defined abnormalities without a definite tumor mass on neuroimaging, the diagnosis can be difficult. We describe a case in which magnetic resonance spectroscopy (MRS) findings suggested a tumor and supported the decision to perform biopsy of the lesion. PMID- 16700049 TI - Exploring structurally conserved solvent sites in protein families. AB - Protein-bound water molecules are important components of protein structure, and therefore, protein function and energetics. Although structural conservation of solvent has been studied in a few protein families, a lack of suitable computational tools has hindered more comprehensive analyses. Herein we present a semiautomated computational approach for identifying solvent sites that are conserved among proteins sharing a common three-dimensional structure. This method is tested on six protein families: (1) monodomain cytochrome c, (2) fatty acid binding protein, (3) lactate/malate dehydrogenase, (4) parvalbumin, (5) phospholipase A2, and (6) serine protease. For each family, the method successfully identified previously known conserved solvent sites. Moreover, the method discovered 22 novel conserved solvent sites, some of which have higher degrees of conservation than the previously known sites. All six families studied had solvent sites with more than 90% conservation and these sites were invariably located in regions of the protein with very high sequence conservation. These results suggest that highly conserved solvent sites, by virtue of their proximity to conserved residues, should be considered as one of the defining three dimensional structural characteristics of protein families and folds. PMID- 16700045 TI - Longitudinal growth and risk factors for growth deficiency in children treated for acute lymphoblastic leukemia. AB - BACKGROUND: Growth deficit has been reported as a frequent complication of the treatment of acute lymphoblastic leukemia (ALL). PROCEDURE: Longitudinal analysis of the growth of 129 children, from a total of 351 cases diagnosed between 1987 and 1994 in Brazil, was determined. Height data were converted into standard deviation Z scores. Only girls younger than 10 and boys younger than 12 years old at diagnosis were included. Patients were treated according to a German BFM-83 based protocol. Fifty-eight children received 18 Gy cranial irradiation, four 12 Gy, and two 24 Gy. Patients were aggregated into five non-excluding groups according to availability of height data at diagnosis, during the treatment, at the end of it, and several years after; 35 children reached their final height. RESULTS: Height deficit at the end of the therapeutic treatment was evident (P < 0.0001). Catch-up occurred 1 year after stopping treatment (P = 0.016). At the last follow-up, over 5 years after the end of treatment (n = 83) or at final height (n = 35), impressive height deficits were recorded (P < 0.0001 for both end points). Multivariate analysis demonstrated that growth impairment was more severe in children younger than 4 years at diagnosis and in those who received cranial irradiation. No significant effect of gender was observed. Children who were treated solely with chemotherapy also had significant height loss. CONCLUSIONS: Treatment of ALL in children is associated with growth deficit during the treatment and several years after it, affecting the final height negatively, particularly in patients younger than 4 and in those who received cranial irradiation. PMID- 16700050 TI - The crystal structure of hypothetical methyltransferase from Thermus thermophilus HB8. PMID- 16700051 TI - Small cationic protein from a marine turtle has beta-defensin-like fold and antibacterial and antiviral activity. AB - Egg white of marine turtle Caretta caretta contains a small cationic protein but lacks lysozyme. The protein was sequenced by a combination of sequential Edman degradation, carboxypeptidase digestion, nuclear magnetic resonance (NMR) and electrospray ionization tandem mass spectrometry. The protein contains 36 amino acid residues of which six are half-cysteines. The three-dimensional structure of the protein was deduced from two-dimensional NMR experiments and was observed to be similar to vertebrate beta-defensins. However, disulfide connectivity is C1 C6/C2-C5/C3-C4; different from that of the vertebrate beta-defensins. The protein showed strong antibacterial activity against Escherichia coli and Salmonella typhimurium. The protein also showed significant antiviral activity against an enveloped rhabdovirus, Chandipura virus, which is an emerging human pathogen. This virus is also closely related to the vesicular stomatitis virus, whose growth was also inhibited. This small cationic protein is part of the innate immunity of this organism and replaces lysozyme in the egg. It has the potential to be developed as an antibacterial and antiviral agent. PMID- 16700052 TI - Blepharophimosis-mental retardation (BMR) syndromes: A proposed clinical classification of the so-called Ohdo syndrome, and delineation of two new BMR syndromes, one X-linked and one autosomal recessive. AB - We report on 11 patients from 8 families with a blepharophimosis and mental retardation syndrome (BMRS) phenotype. Using current nosology, five sporadic patients have Ohdo syndrome, associated with congenital hypothyroidism in two of them (thus also compatible with a diagnosis of Young-Simpson syndrome). In two affected sibs with milder phenotype, compensated hypothyroidism was demonstrated. In another family, an affected boy was born to the unaffected sister of a previously reported patient. Finally, in the last sibship, two affected boys in addition had severe microcephaly and neurological anomalies. A definitive clinical and etiologic classification of BMRS is lacking, but closer phenotypic analysis should lead to a more useful appraisal of the BMRS phenotype. We suggest discontinuing the systematic use of the term "Ohdo syndrome" when referring to patients with BMRS. We propose a classification of BMRS into five groups: (1) del(3p) syndrome, (possibly overlooked in older reports); (2) BMRS, Ohdo type, limited to the original patients of Ohdo; (3) BMRS SBBYS (Say Barber/Biesecker/Young-Simpson) type, with distinctive dysmorphic features and inconstant anomalies including heart defect, optic atrophy, deafness, hypoplastic teeth, cleft palate, joint limitations, and hypothyroidism. BMRS type SBBYS is probably an etiologically heterogeneous phenotype, as AD and apparently AR forms exist; (4) BMRS, MKB (Maat-Kievit-Brunner) type, with coarse, triangular face, which is probably sex-linked; (5) BMRS V (Verloes) type, a probable new type with severe microcephaly, hypsarrhythmia, adducted thumbs, cleft palate, and abnormal genitalia, which is likely autosomal recessive. Types MKB and V are newly described here. PMID- 16700053 TI - Autistic behavior in children with fragile X syndrome: prevalence, stability, and the impact of FMRP. AB - We examined autistic behavior in a cross-sectional sample of 179 children with fragile X syndrome (FXS) and a longitudinal subset of 116 children using the Childhood Autism Rating Scale (CARS) to (a) determine a prevalence of autistic behavior in FXS, (b) examine the stability of autistic ratings over time, and (c) assess the association between the fragile X mental retardation protein (FMRP) and autistic behavior. Approximately 21% of the sample of 129 children (25.9% of boys) scored at or above the cutoff for autism. CARS scores increased slowly, yet significantly, over time, and low levels of FMRP were associated with higher mean levels of autistic behavior as measured by the CARS. PMID- 16700054 TI - Epilepsy and deletions at chromosome 2q24. PMID- 16700055 TI - Ontogeny of the skull, lower jaw, and hyobranchial skeleton of Hypogeophis rostratus (Amphibia: Gymnophiona: Caeciliidae) revisited. AB - Few detailed descriptions of the development of the head skeleton in caecilian amphibians are available. One of those is the work of Marcus and students (e.g., Gehwolf [1923] Z Anat Entwick 68:433-454; Marcus [1933] Anat Anz 80:142-146; Marcus et al. [1935] Gegenbaurs Morphol Jb 76:375-420) on the morphology and development of the skull, lower jaw, and hyobranchial skeleton in the Seychellean caeciliids Hypogeophis rostratus and Grandisonia alternans. These workers described a high number of individual ossifications that fuse during ontogeny to form the adult skull. Although later studies have doubted the generality of those observations, the work of Marcus and his students has been hugely influential in subsequent studies of caecilian skull morphology and amphibian evolution. Based on new observations on an ontogenetic series of 32 sectioned and cleared and stained specimens, ranging from the beginning of chondrification to the adult, the development of the skull, lower jaw, and hyobranchial skeleton of H. rostratus are described. The new results are largely incompatible with those of Marcus and students and no evidence for several of the reported ossifications, including supra-, infra-, and basioccipital, epiotic, pleurosphenoid, preethmoid, posterior vomer, prepalatine, quadratojugal, postparietal, second coronoid, supraangular, and complementare, is found. It is argued that most of Marcus et al.'s reports of nonexistent ossifications are based on false phylogenetic preconception, misinterpretation of the observed morphology, and technical error. Data on the ossification sequence of the skull and lower jaw in H. rostratus are provided and briefly compared to published information on Dermophis mexicanus and Gegeneophis ramaswamii. PMID- 16700056 TI - Local feature analysis: a statistical theory for reproducible essential dynamics of large macromolecules. AB - Multivariate statistical methods are widely used to extract functional collective motions from macromolecular molecular dynamics (MD) simulations. In principal component analysis (PCA), a covariance matrix of positional fluctuations is diagonalized to obtain orthogonal eigenvectors and corresponding eigenvalues. The first few eigenvectors usually correspond to collective modes that approximate the functional motions in the protein. However, PCA representations are globally coherent by definition and, for a large biomolecular system, do not converge on the time scales accessible to MD. Also, the forced orthogonalization of modes leads to complex dependencies that are not necessarily consistent with the symmetry of biological macromolecules and assemblies. Here, we describe for the first time the application of local feature analysis (LFA) to construct a topographic representation of functional dynamics in terms of local features. The LFA representations are low dimensional, and like PCA provide a reduced basis set for collective motions, but they are sparsely distributed and spatially localized. This yields a more reliable assignment of essential dynamics modes across different MD time windows. Also, the intrinsic dynamics of local domains is more extensively sampled than that of globally coherent PCA modes. PMID- 16700057 TI - Proposal of a novel diabetogenic mechanism involving the serpin PAI-1. AB - Metabolic Syndrome is a cluster of risk factors (including obesity, hypertension and insulin resistance), which is associated with late-onset diabetes and coronary heart disease. Elevated levels of the protease inhibitor PAI-1 are well known molecular markers of the Metabolic Syndrome. Here, however, we present a hypothesis that PAI-1 acts as a causative factor in the development of Metabolic Syndrome and its clinical sequelae. We propose that PAI-1 inhibits the activity of members of the proprotein convertase (PC) class of serine proteases and that this underlies, at a molecular level, many of the other features of the Metabolic Syndrome cluster. PMID- 16700058 TI - Interview with professor James Crow. PMID- 16700059 TI - Divided loyalties: transdetermination and the genetics of tissue regeneration. AB - Most tissues contain cells capable of the self-renewal and differentiation necessary to maintain tissue and organ integrity. These somatic stem cells are generally thought to have limited developmental potential. The mechanisms that restrict cell fate decisions in somatic stem cells are only now being understood. This understanding will be important in the clinical exploitation of adult stem cells in tissue repair and replacement. Experiments performed over fifty years ago in Drosophila showed that developmental restriction could be relaxed in the proliferating larval cells that are destined to form the adult fly integument. This phenomenon, called transdetermination, can serve as a model for mechanisms of stem-cell commitment. A recent publication (1) sheds new light on the mechanism of transdetermination by demonstrating that loss of homeotic gene silencing leads to increased frequency of transdetermination. In addition, the authors link a specific signaling pathway induced by tissue regeneration to the relaxation of homeotic gene silencing. The data identify key mechanisms that control developmental homeostasis and cell fate restriction that could be manipulated to make somatic stem-cell engineering possible. PMID- 16700060 TI - A golden clue to human skin colour variation. AB - Variations in human skin pigmentation are obvious, but how have skin colour differences evolved? Although clearly a polymorphic trait, the number and identity of key variants has remained unclear. Investigation of pigmentation phenotypes in model organisms provides a route to identify these genes and showed MC1R to be one key locus. Now, cloning of a classic zebrafish mutant, golden, identifies slc24a5 as a gene involved in fish skin pigmentation.1 Strikingly this study identifies the human orthologue, SLC24A5, as likely to make a major contribution to the pale skin colouration of Western Europeans. PMID- 16700061 TI - Evolution of the mammary gland from the innate immune system? AB - The mammary gland is a skin gland unique to the class Mammalia. Despite a growing molecular and histological understanding of the development and physiology of the mammary gland, its functional and morphological origins have remained speculative. Numerous theories on the origin of the mammary gland and lactation exist. The purpose of the mammary gland is to provide the newborn with copious amounts of milk, a unique body fluid that has a dual role of nutrition and immunological protection. Interestingly, antimicrobial enzymes, such as xanthine oxidoreductase or lysozyme, are directly involved in the evolution of the nutritional aspect of milk. We outline that xanthine oxidoreductase evolved a dual role in the mammary gland and hence provide new evidence supporting the hypothesis that the nutritional function of the milk evolved subsequent to its protective function. Therefore, we postulate that the mammary gland evolved from the innate immune system. In addition, we suggest that lactation partly evolved as an inflammatory response to tissue damage and infection, and discuss the observation that the two signaling pathways, NF-kB and Jak/Stat, play central roles in inflammation as well as in lactation. PMID- 16700062 TI - Commonalities in compensation. AB - The sex chromosomes of many species differ in dosage but the total gene expression output is similar, a phenomenon referred to as dosage compensation. Previously, diverse mechanisms were postulated to account for compensation in distantly related taxa. However, two recent papers present evidence that dosage compensation in Drosophila, mammals and nematodes share the property that there is an approximately two-fold upregulation of the single active X chromosome in each case.(1,2) The results suggest that a common mechanism might operate in these different cases. PMID- 16700063 TI - The dual nature of neurotrophins. AB - Neurotrophins are a small family of dimeric secretory proteins in vertebrate neurons with a broad spectrum of functions. They are generated as pro-proteins with a functionality that is distinct from the proteolytically processed form. The cellular responses of neurotrophins are mediated by three different types of receptor proteins, the receptor tyrosine kinases of the Trk family, the neurotrophin receptor p75(NTR), which is a member of the tumor necrosis factor receptor (TNFR) superfamily, and sortilin, previously characterized as neurotensin receptor. Recent studies have revealed an intriguing pattern: neurotrophins can elicit opposing signals utilising their variable configuration and different receptor types. PMID- 16700064 TI - Mitochondrial one-carbon metabolism is adapted to the specific needs of yeast, plants and mammals. AB - In eukaryotes, folate metabolism is compartmentalized between the cytoplasm and organelles. The folate pathways of mitochondria are adapted to serve the metabolism of the organism. In yeast, mitochondria support cytoplasmic purine synthesis through the generation of formate. This pathway is important but not essential for survival, consistent with the flexibility of yeast metabolism. In plants, the mitochondrial pathways support photorespiration by generating serine from glycine. This pathway is essential under photosynthetic conditions and the enzyme expression varies with photosynthetic activity. In mammals, the expression of the mitochondrial enzymes varies in tissues and during development. In embryos, mitochondria supply formate and glycine for purine synthesis, a process essential for survival; in adult tissues, flux through mitochondria can favor serine production. The differences in the folate pathways of mitochondria depending on species, tissues and developmental stages, profoundly alter the nature of their metabolic contribution. PMID- 16700065 TI - Regulation of functional diversity within the Nedd4 family by accessory and adaptor proteins. AB - Ubiquitination is essential in mediating diverse cellular functions including protein degradation and trafficking. Ubiquitin-protein (E3) ligases determine the substrate specificity of the ubiquitination process. The Nedd4 family of E3 ligases is an evolutionarily conserved family of proteins required for the ubiquitination of a large number of cellular targets. As a result, this family regulates a wide variety of cellular processes including transcription, stability and trafficking of plasma membrane proteins, and the degradation of misfolded proteins. The modular architecture of the proteins, comprising a C2 domain, multiple WW domains and a catalytic domain, enables diverse intermolecular interactions and recruitment to various subcellular locations. The WW domains commonly mediate interaction with substrate proteins; however, an increasing number of Nedd4 targets do not contain obvious WW domain-interaction motifs suggesting the involvement of accessory proteins. This review discusses recent insights into how accessory and adaptor proteins modulate the activities of Nedd4 family members, including recruitment of novel substrates, alteration of subcellular localisation and effects on ubiquitination. PMID- 16700066 TI - The ovotestis: an underdeveloped organ of evolution. AB - In animals that have separate sexes (gonochorists), many sperm are produced to fertilise a few eggs. As the male germline undergoes more mitoses, so the accumulated mutation frequency is elevated in sperm compared with ova, and evolution is 'male-driven'. In contrast, in many hermaphroditic animals, a single organ--the ovotestis--produces both ova and sperm. Since self-renewing cells in the ovotestis may give rise to both cell types throughout life, ova in hermaphrodites could in theory have undergone as many cell divisions as sperm. Here, I consider some possible effects of the ovotestis on evolution. In particular, I hypothesise that the accumulated mutation frequency of nuclear genes in hermaphrodites (including species that change sex) may reach twice that compared with gonochorists. There may be an even greater increase in the mitochondrial mutation frequency. Further developmental studies and the accumulation of comparative data should allow hypothesis testing. If the prediction is correct, then it may provide the most-straightforward explanation for the extraordinary diversity of mitochondrial DNA in some hermaphrodites, especially molluscs. PMID- 16700067 TI - Pristionchus pacificus: a well-rounded nematode. AB - Nematodes pervade Earth's biosphere and occupy innumerable ecological niches. The role of Caenorhabditis elegans as a model for developmental processes has encouraged us to cultivate a second nematode, Pristionchus pacificus, as a comparative counterpoint to address questions in development, behavior and ecology in nematode evolution. We hope that this endeavor, now more than a decade underway, will allow us to project findings onto other comparative models for biological processes. To this end, our laboratory has made an extensive genetic map and mutant screens to understand changes in developmental programs. Recently, we have been capitalizing on the whole genome sequence of P. pacificus to describe more thoroughly the molecular basis for these changes, as well as to better integrate our molecular knowledge with the biodiversity of Pristionchus species. PMID- 16700068 TI - The complete genome sequence of a dog: a perspective. AB - A complete, high-quality reference sequence of a dog genome was recently produced by a team of researchers led by the Broad Institute, achieving another major milestone in deciphering the genomic landscape of mammalian organisms. The genome sequence provides an indispensable resource for comparative analysis and novel insights into dog and human evolution and history. Together with the survey sequence of a poodle previously published in 2003, the two dog genome sequences allowed identification of more than 2.5 million single nucleotide polymorphisms within and between dog breeds, which can be used in evolutionary analysis, behavioral studies and disease gene mapping.(1) PMID- 16700070 TI - Culturing in vitro produced blastocysts in sequential media promotes ES cell derivation. AB - Embryonic stem (ES) cell lines are routinely derived from in vivo produced blastocysts. We investigated the efficiency of ES cells derivation from in vitro produced blastocysts either in monoculture or sequential culture. Zygotes from hybrid F1 B6D2 mice were cultured in vitro to the blastocyst stage in Potassium (K(+)) simplex optimised medium (KSOM) throughout or in KSOM and switched to COOK blastocyst medium on day 3 (KSOM-CBM). Blastocysts were explanted on a feeder layer of mitomycin C-inactivated murine embryonic fibroblasts (MEF) in TX-WES medium for ES cell derivation. Sequential KSOM-CBM resulted in improved blastocyst formation compared to KSOM monoculture. ES cells were obtained from 32.1% of explanted blastocsyts cultured in KSOM-CBM versus 18.4% in KSOM alone. ES cell lines were characterized by morphology, expression of SSEA-1, Oct-4 and alkaline phosphatase activity, and normal karyotype. These results indicate that in vitro culture systems to produce blastocysts can influence the efficiency of ES cell line derivation. PMID- 16700069 TI - Addition of glutathione or thioredoxin to culture medium reduces intracellular redox status of porcine IVM/IVF embryos, resulting in improved development to the blastocyst stage. AB - The present series of experiments investigated the effect of a reducing environment created by addition of reduced glutathione (GSH) or thioredoxin (TRX) to in vitro culture medium on the developmental competence of in vitro produced porcine embryos, and their intracellular redox status. Porcine cumulus-oocyte complexes were collected from ovaries matured and fertilized in vitro. The putative zygotes were then cultured for 6 days in modified NCSU-37 medium with or without (control) GSH or TRX, and their developmental competence was evaluated. In addition, the intracellular redox status of the cultured embryos was compared quantitatively using an index based on the ratio of the intracellular GSH content relative to the intracellular H(2)O(2) level. The proportion of embryos that developed to the blastocyst stage was significantly increased when 0.5 or 1.0 microM GSH (29.6% or 30.4%, P < 0.05 or 0.01, respectively) or 1.0 mg/ml TRX (30.6%, P < 0.01) was added to the medium compared to that without any supplementation (control; 20.1%). The intracellular redox status of embryos at the 8- to 12-cell stage or the blastocyst stage in the group cultured in the presence of GSH or TRX was significantly reduced in comparison with the control (P < 0.05 to 0.001). Furthermore, administration of GSH or TRX enhanced the total cell number (from 48.3 to 49.2) and lowered the proportion of apoptotic cells (from 6.2% to 7.0%) in blastocysts compared with the control (cell number 39.3; apoptosis rate 11.1%, P < 0.05). These results suggest that GSH or TRX can improve the in vitro development of porcine embryos, while maintaining an intracellular reductive status. PMID- 16700071 TI - Spermatocyte responses in vitro to induced DNA damage. AB - Spermatocytes normally sustain many meiotically induced double-strand DNA breaks (DSBs) early in meiotic prophase; in autosomal chromatin, these are repaired by initiation of meiotic homologous-recombination processes. Little is known about how spermatocytes respond to environmentally induced DNA damage after recombination-related DSBs have been repaired. The experiments described here tested the hypothesis that, even though actively completing meiotic recombination, pachytene spermatocytes cultured in the absence of testicular somatic cells initiate appropriate chromatin remodeling and cell-cycle responses to environmentally induced DNA damage. Two DNA-damaging agents were employed for in vitro treatment of pachytene spermatocytes: gamma-irradiation and etoposide, a topoisomerase II (TOP2) inhibitor that results in persistent unligated DSBs. Chromatin modifications associated with DSBs were monitored after exposure by labeling surface-spread chromatin with antibodies against RAD51 (which recognizes DSBs) and the phosphorylated variant of histone H2AFX (herein designated by its commonly used symbol, H2AX), gammaH2AX (which modifies chromatin associated with DSBs). Both gammaH2AX and RAD51 were rapidly recruited to irradiation- or etoposide-damaged chromatin. These chromatin modifications imply that spermatocytes recruit active DNA damage responses, even after recombination is substantially completed. Furthermore, irradiation-induced DNA damage inhibited okadaic acid-induced progression of spermatocytes from meiotic prophase to metaphase I (MI), implying efficacy of DNA damage checkpoint mechanisms. Apoptotic responses of spermatocytes with DNA damage differed, with an increase in frequency of early apoptotic spermatocytes after etoposide treatment, but not following irradiation. Taken together, these results demonstrate modification of pachytene spermatocyte chromatin and inhibition of meiotic progress after DNA damage by mechanisms that may ensure gametic genetic integrity. PMID- 16700072 TI - Sexually dimorphic expression of secreted frizzled-related (SFRP) genes in the developing mouse Mullerian duct. AB - In developing male embryos, the female reproductive tract primordia (Mullerian ducts) regress due to the production of testicular anti-Mullerian hormone (AMH). Because of the association between secreted frizzled-related proteins (SFRPs) and apoptosis, their reported developmental expression patterns and the role of WNT signaling in female reproductive tract development, we examined expression of Sfrp2 and Sfrp5 during development of the Mullerian duct in male (XY) and female (XX) mouse embryos. We show that expression of both Sfrp2 and Sfrp5 is dynamic and sexually dimorphic. In addition, the male-specific expression observed for both genes prior to the onset of regression is absent in mutant male embryos that fail to undergo Mullerian duct regression. We identified ENU-induced point mutations in Sfrp5 and Sfrp2 that are predicted to severely disrupt the function of these genes. Male embryos and adults homozygous for these mutations, both individually and in combination, are viable and apparently fertile with no overt abnormalities of reproductive tract development. PMID- 16700074 TI - Antioxidant status of the lower oviduct in the chicken varies with age and dietary vitamin E supplementation. AB - Protection of sperm membranes against lipid peroxidation is a pre-requisite to prolonged sperm storage, both in vivo and in vitro. As females from avian species can store spermatozoa in the utero-vaginal junction (UVJ) for prolonged periods, we investigated the mechanisms involved in antioxidative protection of the plasma membrane of chicken sperm in this region. Comparisons of concentrations in nonenzymatic (alpha-tocopherol, ascorbic acid, and GSH) and enzymatic (GSH-Px, SOD) antioxidants among the vagina, UVJ and uterus of sexually mature chicken hens revealed tissue-specific profiles, with higher ascorbic acid content and increased GSH-Px and SOD activity in the UVJ compared to other regions of the lower oviduct (vagina, uterus). Deterioration of the antioxidant profile in the UVJ was observed in aging hens, but it was partially compensated by dietary supplementation with vitamin E (130 ppm). It is concluded that the chicken UVJ provides a complex defense barrier against lipid peroxidation of the sperm membrane during in vivo storage, which can be partially improved by dietary supplementation with vitamin E. The protective effects of this barrier decline over time during the reproductive season. PMID- 16700073 TI - Characterization of the mRNA expression of StAR and steroidogenic enzymes in zebrafish ovarian follicles. AB - The objective of this study was to investigate the levels of expression of steroid biosynthetic enzymes and steroidogenic acute regulatory protein (StAR) at different stages of ovarian follicular development in zebrafish (Danio rerio), and to investigate the sites within the steroid biosynthetic pathway that may be regulated by gonadotropins. Ovarian follicles of sexually mature fish were separated into primary, previtellogenic, vitellogenic, and mature stages and the expression of StAR, P450 side chain cleavage (P450scc), 3beta-hydroxysteroid dehydrogenase (3beta-HSD), P450 hydroxylase/lyase (P450c17), 17beta hydroxysteroid dehydrogenase type 1 (17beta-HSD1), 17beta-hydroxysteroid dehydrogenase type 3 (17beta-HSD3), and P450 aromatase (P450aromA) was determined by Real time RT-PCR. The expression of all genes changed significantly as follicles grew, with a decrease in the expression of StAR, P450scc, 3beta-HSD and P450c17 with maturation, and an increase in the expression of 17beta-HSD3 during vitellogenesis and 17beta-HSD1 and P450aromA during previtellogenesis. In vitro incubation of vitellogenic follicles demonstrated that the expression of StAR, 17beta-HSD3, and P450aromA increased in response to hCG, and decreased in the absence of hCG. In contrast, the expression of P450scc, 3beta-HSD, P450c17, and 17beta-HSD1 remained constant between treatments and over time. Testosterone and estradiol production in the culture medium was stimulated by human chorionic gonadotropin (hCG). These experiments aid in the characterization of the roles and regulation of steroids throughout ovarian development, and suggest that gonadotropins play a key role in the regulation of StAR, 17beta-HSD3, and P450aromA in zebrafish. PMID- 16700075 TI - Expression of the spermatogenic cell-specific glyceraldehyde 3-phosphate dehydrogenase (GAPDS) in rat testis. AB - The spermatogenic cell-specific variant of glyceraldehyde 3-phosphate dehydrogenase (GAPDS) has been cloned from a rat testis cDNA library and its pattern of expression determined. A 1,417 nucleotide cDNA has been found to encode an enzyme with substantial homology to mouse GAPDS (94% identity) and human GAPD2 (83% identity) isozymes. Northern blotting of rat tissue RNAs detected the 1.5 kb Gapds transcript in the testis and not in RNA from liver, spleen, epididymis, heart, skeletal muscle, brain, seminal vesicle, and kidney. The rat Gapds mRNA was first detected at day 29 of postnatal testis development, an age which coincides with the initial post-meiotic differentiation of round spermatids. When isolated rat spermatogenic cell RNA was probed for Gapds expression, transcripts were detected only in round spermatids and condensing spermatids, but not in pachytene spermatocytes, demonstrating haploid expression of the Gapds gene. However, immunohistochemical staining of rat testis sections with anti-GAPDS antisera did not detect GAPDS in round spermatids, but localized the protein only to stage XIII and later condensing spermatids as well as testicular spermatozoa, indicating that Gapds expression is translationally regulated. The current results are similar to those previously obtained for mouse GAPDS and human GAPD2, suggesting that reliable comparisons can be made between these species in toxicant screening and contraceptive development. PMID- 16700076 TI - Steroid hormones acutely regulate expression of a Nudix protein-encoding gene in the endometrial epithelium of sheep. AB - Steroid hormones regulate endometrial gene expression to meet the needs of developing embryos. Our hypothesis is that steroid hormones transiently induce expression of genes in the endometrial epithelium to make the uterine environment different between the earliest days of pregnancy. We identified one such gene product using differential display-polymerase chain reactions. The gene product that was strongly induced in ewes between day 3 and 6 of the estrous cycle was cloned and sequenced to identify it as encoding a member of the Nudix family of hydrolase enzymes. Northern blot analyses indicated that NUDT16 mRNA concentrations were elevated 10-fold in the endometrium of sheep from day 5 to 9 of the estrous cycle and returned to basal levels by day 11. In assays of RNA samples from 15 different tissues from an adult ewe, the concentrations of NUDT16 mRNA were greatest in endometrium. In situ hybridization localized NUDT16 mRNA exclusively to the endometrial epithelial cells of the glands and uterine lumen. In ovariectomized ewes, NUDT16 mRNA was induced by a regimen of alternating estrogen and progesterone therapy designed to mimic the hormonal experiences of a ewe at day 6 of the estrous cycle. The final estrogen treatment in the regimen was critical to the expression of NUDT16 as well as progesterone receptor and estrogen receptor-beta genes. Characterization of the NUDT16 gene identified putative steroid hormone response elements, which can now be investigated to understand its unique pattern of regulation in the earliest days of pregnancy. PMID- 16700077 TI - Anxiolytics, hypnotics, and antidepressants dispensed to adolescents in a French region in 2002. AB - PURPOSE: This study proposes to complete declarative studies by describing the prescriptions of anxiolytics, hypnotics, and antidepressants dispensed to adolescents in a French region in 2002. METHODS: This cross-sectional study analyzes all the hypnotic, anxiolytic, and antidepressant prescriptions (ATC codes beginning with N05B, N05C, and N06A, respectively) sent by adolescents (aged 13-17 years) to the French Health Insurance system of the study region for reimbursement during one year (2002). It was performed in a southern France area with 120,908 adolescents covered by this insurance scheme. Adverse drug reactions (ADRs) recorded in the Pharmacovigilance database were also studied. RESULTS: Three thousand two hundred and eighty-six adolescents (2.7% of adolescent population) had at least one prescription of the studied drugs. This prevalence increased with age and female sex, leading to a maximum of 6.3% for the 17-year old girls. Two thousand four hundred and thirty-one of adolescents were dispensed anxiolytics, 935 antidepressants, and 548 hypnotics. The most dispensed drugs were zolpidem, zopiclone, and niaprazine for hypnotics; hydroxyzine, etifoxine, and bromazepam for anxiolytics; and paroxetine, sertraline, and fluoxetine for antidepressants. Zolpidem, hydroxyzine, and paroxetine accounted, respectively, for 82.9%, 57.1%, and 59.8% of the prescriptions. 75.5% of hypnotics users had only one prescription, 77.4% for anxiolytics, and 57.4% for antidepressants. Three ADRs were reported. CONCLUSIONS: This study confirms the large use of psychotropics in French adolescents and the influence of age and sex. Also, the results underline treatment for most adolescents is short, which may be beneficial for hypnotics and anxiolytics but not for antidepressants. PMID- 16700078 TI - Quality of manufacturer provided information on safety and efficacy claims for dietary supplements for colonic health. AB - PURPOSE: Food and Drug Administration (FDA) approval of dietary supplements is not required. However, manufacturers must ensure that data exist to support safety and efficacy claims of their product. This study was designed to evaluate accessibility and quality of such data from manufacturers of dietary supplements for 'colonic health.' METHODS: Supplements promoting 'colonic health' were identified at area stores. A physician contacted the manufacturers by mail and telephone requesting data to substantiate claims of efficacy and safety. MEDLINE was searched to identify reports of adverse events or medication interactions. RESULTS: Twelve manufacturers of 23 products were surveyed. Eight manufacturers responded, of whom four provided no clinical data to substantiate claims of efficacy or safety. No manufacturer provided data that directly evaluated their product. Our literature review identified a wide range of potential adverse events and drug-supplement interactions, albeit mostly as case reports, animal studies, or in vitro experiments. CONCLUSIONS: There is a need to increase availability of supplement safety data to physicians and the general public. Consideration should be given to various responses, including legislative actions that address this issue. PMID- 16700079 TI - Antibiotic use in ambulatory care in Europe (ESAC data 1997-2002): trends, regional differences and seasonal fluctuations. AB - PURPOSE: The ESAC project (European Study on Antibiotic Consumption) aims to collect antibiotic-use data through a European network of national surveillance systems. This paper reports on the retrospective data collection in ambulatory care for the period 1997-2002. METHODS: Valid data of antibiotic consumption of 24 European countries for 2002 and of 18 countries for the entire 6-year period was classified according to the Anatomical Therapeutic Chemical Classification (ATC) and expressed in defined daily dose (DDD) per 1000 inhabitants per day (DID). Overall and subgroup comparison of antibiotic consumption over time as well as between geographical clusters was performed. RESULTS: Total use of antibiotics in Europe remained at a median level of 20 DID in the period 1997 2002 with a wide variation between countries ranging from 9.8 DID in The Netherlands to 32.2 DID in France. A substantial increase in subclass consumption of co-amoxiclav and fluoroquinolones was noted while the use of narrow-spectrum penicillins, erythromycin, quinolones and sulfonamides decreased. Total consumption as well as seasonal fluctuations showed remarkable geographical clustering with low consumption and low variation between summer and winter in the North, high consumption patterns in the South and a mixed model in the East. CONCLUSIONS: Within the ESAC project, valid time series of antibiotic-use data are publicly available now, enabling to improve the study of determinants of use, the evaluation of governmental antibiotic consumption policies and the investigation of the associated emergence of antibiotic resistance. PMID- 16700080 TI - Changes in use of disease-modifying anti-rheumatic drugs in Australia over the period 1992-2004. AB - PURPOSE: Evidence is growing that early use of disease-modifying anti-rheumatic drugs (DMARDs) and combinations of these drugs provide optimal care for people with rheumatoid arthirits. The aim of this study was to describe objectively the pattern of consumption of DMARDs in the Australian community (community-based prescribing, specialist and general practitioner) 1992-2004, and to compare this with prescribing patterns reported in other countries. METHOD: Dispensing statistics from the Pharmaceutical Benefit Scheme (PBS-Australia's universal prescription subsidy scheme) were analysed and temporal trends evaluated. Drug consumption was calculated as the number of dispensed defined daily doses (DDD)/1000 inhabitants/day (WHO ATC/DDD classification 2005). RESULTS: The consumption of DMARDs in the Australian community increased steadily from 2.6 DDD/1000 inhabitants/day in 1992 to 5.5 DDD/1000 inhabitants/day. Over the period 1992-2004, methotrexate (MTX) was the most commonly used DMARD (from 0.6 to 3.0 DDD/1000 inhabitants/day). Consumption of gold (parenteral and oral) and penicillamine declined during this time. The inclusion of leflunomide on the PBS in 2000 contributed to the increase in DMARD usage. CONCLUSION: Use of DMARDs within the Australian community has increased in recent years, coinciding with the change in guidelines for therapy for rheumatoid arthritis (RA) to earlier use of DMARDs and the more common use of combinations. This study used DDD methodology to quantify trends for DMARD consumption and these trends are broadly consistent with international prescribing patterns assessed using different methodologies. PMID- 16700081 TI - How is leflunomide prescribed and used in Australia? Analysis of prescribing and adverse effect reporting. AB - PURPOSE: To evaluate the use of leflunomide in the Australian community since introduction in 2000. Trends in adverse drug reaction (ADR) reporting were also studied. METHODS: Annual Australian prescription and dispensing statistics were analysed. Drug utilisation was estimated as defined daily doses (DDD)/1000 inhabitants/day. ADR data from the Therapeutic Goods Administration's Adverse Drug Reactions Advisory Committee (ADRAC) national monitoring system were compared with the World Health Organisation (WHO) Vigibase records. RESULTS: Leflunomide use in Australia (dispensing data) increased from 0.2 in 2000 to 0.4 DDD/1000 inhabitants/day in 2002. The same overall pattern was observed in the 'authority to prescribe' data. From 2000-2002, prescribing of the starter pack (3 x 100 mg loading dose plus 30 x 20 mg tablets) declined (down 74%); likewise for the 20 mg (30 tablets) pack. Gradual increases were noted for the 10 mg (30 tablets) pack (up 40%). Approximately 135 reports, detailing about 370 individual ADR, were generated annually. Gastro-intestinal disorders predominated, accounting for 24% of reactions reported to ADRAC. Skin and appendages disorders constituted 14% of reported reactions. Deaths in leflunomide users were attributed to a combination of haematological and gastro-intestinal complications, but it was not possible to ascertain other medication usage or contributing factors. Trends observed with the ADRAC reports were consistent with the WHO database. CONCLUSIONS: Leflunomide was the first registered DMARD in Australia in over a decade and its use has increased within the community. The ADR reports might have contributed to Australian rheumatologists gradually abandoning loading patients with high doses of leflunomide in favour of starting therapy at lower doses. PMID- 16700082 TI - The nature of the scientific evidence leading to drug withdrawals for pharmacovigilance reasons in France. AB - INTRODUCTION: Because of design, objectives and number of included subjects, clinical studies are insufficient to assess the safety of new drugs. Sometimes, serious adverse drug reactions (ADRs) led to withdrawal of the drug from the market after their approval. The objective of our study was to determine the scientific evidences leading to drug withdrawal for pharmacovigilance reasons in France. METHODS: Data coming from French Health Products Safety Agency, literature and Toulouse Pharmacovigilance Center allowed to identify all drugs withdrawn from the French market for pharmacovigilance reasons from 1998 to 2004. We classified data according to their study design (Randomized Clinical Trial [RCT], case serie or case report, case-control study, cohort study, observational study, animal study), the organ/system affected and the type of ADR. RESULTS: A total of 21 drugs were withdrawn for safety reasons between 1998 and 2004 in France. The most frequent ADRs were hepatic (n = 7), cardiovascular (n = 4) or neurological (n = 3) ones. Eleven withdrawals were due to type-B ('unexpected') reactions (52%). For 19 out of 21 drugs, scientific evidence leading to drug withdrawal came from spontaneous case reports (or case series). Among these, case reports were the sole evidence in 12 cases. Withdrawals were based on evidence from case reports in combination with case-control or cohort study in four cases, in combination with observational study in two cases or in combination with animal study in two other cases. In only one case, a RCT supported the decision. CONCLUSIONS: This study underlines the importance of spontaneous case reports in detecting signals and supporting withdrawal of drug for pharmacovigilance reasons in France. Health authorities suffer from lack of comparative data resource. In this perspective, a pharmaco-epidemiological population-based database could represent a helpful tool to both generate and test safety hypotheses. PMID- 16700083 TI - National patterns of medication use during pregnancy. AB - PURPOSE: To describe patterns of medication use during pregnancy in ambulatory care settings according to the U.S. Food and Drug Administration (FDA) pregnancy risk classification. METHODS: A cross-sectional study of two national ambulatory care surveys, sampling all office visits made by pregnant women in 1999 and 2000, was conducted. Using the FDA pregnancy risk classification, patterns of medication use and predictive factors for FDA pregnancy risk D or X (D/X) medications were evaluated. RESULTS: In 1999 and 2000, about half of all pregnant visits had one or more medications. Among the total visits, FDA Class A was the majority (private = 65.7%; hospital = 79.5%; p < 0.05) followed by Class C (private = 26.5%; hospital = 36.4%; p < 0.05). Class D/X medications accounted for 6.4% and 2.9% of visits in private and hospital, respectively (p < 0.05). Medications with unknown pregnancy categories were predominant in the private setting (12.0% and 3.9%; p < 0.05). Age, insurance type, region, physician specialty, and number of medications were associated with a category D/X prescription. Among hospital visits, those from the West region and with private insurance were more likely to receive category D/X prescriptions. Number of medications was strongly associated with high-risk drugs in both settings. CONCLUSIONS: This study shows considerable medication use among pregnant women. The prevalence of visits with FDA pregnancy category D/X drugs was moderate, but still indicates exposure to high-risk medications. PMID- 16700084 TI - Measuring change in prescription drug utilization in Australia. AB - PURPOSE: The National Prescribing Service Ltd (NPS) aims to improve prescribing and use of medicines consistent with evidence-based best practice. This report compares two statistical methods used to determine whether multiple educational interventions influenced antibiotic prescription in Australia. METHODS: Monthly data (July 1996 to June 2003) were obtained from a national claims database. The outcome measures were the median number of antibiotic prescriptions per 1000 consultations for each general practitioner (GP) each month, and the mean proportion (across GPs) of each subgroup of antibiotics (e.g. roxithromycin) out of nine antibiotics having primary use for upper respiratory tract infection. Two approaches were used to investigate shifts in prescribing: augmented regression, which included seasonality, autocorrelation and one intervention; and seasonally adjusted piecewise linear dynamic regression, which removed seasonality prior to modelling, included several interventions, GP participation and autocorrelated errors. Both methods are variations of piecewise linear regression modelling. RESULTS: Both approaches described a similar decrease in rates, with a non significant change after the first intervention. The inclusion of more interventions and GP participation made no difference. Using roxithromycin as an example of the analyses of proportions, both approaches implied that after the first intervention the proportion decreased significantly. The statistical significance of this intervention disappears when other interventions are included. CONCLUSIONS: The two analyses provide results which agree regarding the possible impact of the NPS interventions, but raise questions about what is the best way to model drug utilization, particularly regarding whether to include all intervention terms when they belong to an extended roll-out of related interventions. PMID- 16700085 TI - Co-dispensing of contraindicated medications in patients using cisapride in Italy. AB - PURPOSE: To investigate the co-dispensing of contraindicated medications in patients using cisapride in an Italian population. METHODS: The study included patients who had at least one cisapride prescription dispensed between 1 April 1997 and 30 September 2000. The proportion of cisapride users with concomitant medications, the proportion of prescriptions of cisapride with an overlapping dispensing of a contraindicated medication and the proportion of person-days of cisapride use concomitant with contraindicated medications, were calculated. RESULTS: In the 4-year study period, 249,740 total cisapride prescriptions, which corresponded to 382,835 packages, were dispensed to 91,204 users (52,442 female, 57.5%). Concomitant use of contraindicated drugs was identified in 4403 cisapride users (4.83%). Throughout the study period, there was an increase in the proportion of cisapride users with contraindicated co-prescriptions: 4.29% in 1997, 4.99% in 1998, 5.15% in 1999, but a decline to 3.34% in 2000. In each year, this figure was higher in males, with the highest proportion reported in 1999 (5.50%). A total of 7215 cisapride prescriptions (2.85%) were concomitant with contraindicated drugs, ranging from 2.27% in September 1998 to 3.55% in January 1999. Cisapride prescriptions were most frequently associated with macrolide antibiotics (1.14%) and class III antiarrhythmics (1.03%). CONCLUSIONS: In the period 1997-2000, a substantial proportion of cisapride prescriptions were concomitant with contraindicated medications. In Italy in 1998, the National Health Authority issued a bulletin warning about the risk of serious arrhythmias after concomitant use of contraindicated drugs. Despite this warning, there was no reduction in contraindicated drug uses. PMID- 16700086 TI - Developments of the incidence of osteoporosis in The Netherlands: a PHARMO study. AB - PURPOSE: To assess recent and to predict future time trends in the incidence of osteoporosis using routine databases in The Netherlands in the period 1993-2015 and to compare estimations based on hip fractures versus a proxy based on pharmacy and hospitalisation data. METHODS: The incidence of hip fractures was estimated over the period 1986-2002 using information from the Dutch Medical Registry, covering more than 99% of all Dutch hospitalisations. Additionally, a proxy (hospitalisation for osteoporosis or osteoporotic fractures, treatment with glucocorticosteroids or treatment with anti-osteoporosis drugs) was constructed in order to identify osteoporotic patients in the PHARMO database. Age and gender specific incidences of hip fractures and osteoporosis were calculated and extrapolated to The Netherlands. Results of both studies were extrapolated till 2015 using a power function. RESULTS: The incidence of hip fractures decreased slightly (270 per 100,000 in 1993, 260 per 100,000 in 2002). The incidence of osteoporosis using the constructed proxy decreased from 870 per 100,000 in 1993 to 700 per 100,000 in 2002. The incidence of hip fractures and osteoporosis remained fairly constant when modelled till 2015. Both studies showed the same time trends. CONCLUSION: Both the estimations based on the hospitalisations for hip fracture and on our proxy for osteoporosis showed that the increase in the incidence of osteoporosis as observed in the 1990s is levelling off. Due to ageing of the population the absolute number of hip fractures will however increase. Our definition of osteoporosis resulted in a higher estimation of the incidence of osteoporosis and may be used in future studies to follow developments in osteoporosis prevalence and incidence. PMID- 16700087 TI - Screening in early pregnancy for pre-eclampsia using Down syndrome quadruple test markers. AB - OBJECTIVES: To estimate the screening performance of early second-trimester prenatal serum markers for Down syndrome, in screening for the development of pre eclampsia, and analyse the uncertainty over its screening performance. METHODS: A nested case-control study was carried out on 96 women with pre-eclampsia and 5 controls for each case from among the women attending three hospitals in London for their prenatal care. Record linkage between computerized obstetric and screening databases identified women with pre-eclampsia and unaffected control women. The stored frozen serum samples collected from these pregnancies between 15 and 22 weeks' gestation were retrieved and assayed for alpha-fetoprotein (AFP), unconjugated estriol (uE(3)), total human chorionic gonadotrophin (hCG), free beta-hCG and inhibin-A. RESULTS: Pre-eclampsia was identified from the computerized obstetric records and confirmed by examination of the medical notes. In the pregnancies that went on to develop pre-eclampsia, early second trimester inhibin-A and hCG values were significantly raised and uE(3) values were significantly lowered, while AFP values were not significantly altered. Using the Quadruple test markers (AFP, uE(3), hCG (total or free beta) and inhibin-A), an estimated 34% of pregnancies that developed pre-eclampsia were detected at a 5% false-positive rate. If all the women who had pre-eclampsia in a previous pregnancy (assuming a pre-eclampsia prevalence of 4%) are considered as screen positive and the serum test is applied to the remaining women, then around 42% of pre-eclamptic pregnancies would be detected at a 6.5% false-positive rate. Pre eclampsia screening performance using the Quadruple test markers was materially better than that using the Triple test markers. CONCLUSION: Adding screening for pre-eclampsia to an existing Down syndrome screening programme using the Quadruple test markers is simple and worthwhile. It will detect over 40% of pregnancies with pre-eclampsia at an acceptable false-positive rate (about 6%) and with minimal additional costs. PMID- 16700088 TI - Prenatal diagnosis of de novo deletions of 8p23.1 or 15q26.1 in two fetuses with diaphragmatic hernia and congenital heart defects. AB - OBJECTIVE: To show the importance of using high-resolution chromosome analysis and FISH-technique for finding subtle chromosomal lesions in prenatal diagnosis specially when there are abnormal ultrasound findings. METHODS: Ecographic examination of the fetus. GTG banded chromosome and FISH analysis using subtelomeric probes on amniocytes. RESULTS: We report two prenatal cases with congenital diaphragmatic hernia (CDH) and congenital heart defects (CHDs) with different deletions confirmed by FISH: del(8)(p23.1p23.1) and del(15)(q26.1). CONCLUSION: These cases support the evidence that the regions 15q26.1 and 8p23.1 may play an important role in the development of the diaphragm. A deletion 8p23.1 or 15q26.1 should be considered whenever a CDH and/or a cardiac abnormality are detected on ultrasound. PMID- 16700090 TI - Hartree-Fock energy partitioning in terms of Hirshfeld atoms. AB - A Hirshfeld decomposition scheme of the Hartree-Fock total molecular energy into atomic energies is presented. The calculations are performed by direct numerical integration and the results are compared for a set of 28 molecules containing different kinds of atoms. The calculated atomic energies show a strong dependency on changes of atomic electron population and hybridization. Linear correlations are found between the energy and the population for H, these being related to the electronegativity of this atom and to the external potential created by the remaining atoms. The proposed energy partitioning scheme appears to be useful for studies such as proton acidity, the anomeric effect and group transferability, and allows atomic virial ratios to be obtained. Finally, the atomic potential energies are found to mimic trends based on exact expressions as well as trends displayed by molecular quantities, thus lending credibility to the partitioning scheme used. PMID- 16700089 TI - Selective stabilization of triple-helical DNA by diazoniapolycyclic intercalators. PMID- 16700091 TI - Car-Parrinello molecular dynamics study of the blue-shifted F3CH...FCD3 system in liquid N2. AB - Fluoroform, as confirmed by both experimental and theoretical studies, can participate in improper H-bond formation, which is characterized by a noticeable increase in the fundamental stretching frequency nu(C-H) (so-called blue frequency shift), an irregular change of its integral intensity, and a C-H bond contraction. A Car-Parrinello molecular dynamics simulation was performed for a complex formed by fluoroform (F3CH) and deuterated methyl fluoride (FCD3) in liquid nitrogen. Vibrational analysis based on the Fourier transform of the dipole moment autocorrelation function reproduces the blue shift of the fundamental stretching frequency nu(C-H) and the decrease in the integral intensity. The dynamic contraction of the C-H bond is also predicted. The stoichiometry of the solvated, blue-shifted complexes and their residence times are examined. PMID- 16700092 TI - In vitro effect of adenovirus-mediated human Gamma Interferon gene transfer into human mesenchymal stem cells for chronic myelogenous leukemia. AB - For developing gene therapy for chronic myelogenous leukemia (CML), we evaluated the feasibility of using autologous bone marrow stromal cells (BMSCs) of one CML patient as a target cell population and studied the efficiency of recombinant adenovirus-mediated human Gamma Interferon (hIFN-gamma) gene transfer into BMSCs. BMSCs can be readily obtained, expanded, and successfully transduced with adenoviral vectors in vitro. We studied the in vitro expression of hIFN-gamma in human BMSCs following transduction with Ad/hIFN-gamma. On transduction of BMSCs at a MOI of 50, the expression and secretion of hIFN-gamma were achieved as high as 5492 +/- 660 approximately 50647 +/- 4049 ng/10(6) cells per 24 h over the course of 3 weeks. We further studied the effects of hIFN-gamma produced by transduced BMSCs on the proliferation of the human leukemia cell line K562 cells in vitro, proliferation of K562 cells was markedly inhibited in the experimental groups as compared with the other two control groups after 5 days of coculture. We also found that the percentage of K562 cells in the G(1) phase of cell cycle can be increased by treatment of hIFN-gamma produced by Ad/hIFN-gamma transduced BMSCs, but the percentage of K562 cells in the S phase of cell cycle can be decreased in the same time. Apoptosis rate of K562 cells in the experimental groups was 30.8 +/- 8.5%, as compared with the other two control groups (5.6 +/- 1.3% and 5.5 +/- 0.8%, respectively) (p < 0.01). Our results indicate that hIFN gamma gene engineered BMSCs of CML donors could be successfully established and that local production of hIFN-gamma is sufficiently to inhibit the proliferation of K562 cells and induce apoptosis of K562 cells in vitro, suggesting an important potential use in the clinical gene therapy of CML. PMID- 16700093 TI - Systemic lupus erythematosus prevalence in the UK: methodological issues when using the General Practice Research Database to estimate frequency of chronic relapsing-remitting disease. AB - PURPOSE: The purpose of this study was to calculate the prevalence of systemic lupus erythematosus (SLE) between 1992 and 1998 using the General Practice Research Database (GPRD) METHODS: We identified all individuals who had contributed at least 3 years of data to the GPRD and who had a diagnosis of SLE with supporting evidence of their diagnosis. We calculated the annual age- and sex-specific prevalence of SLE. Additionally, we stratified the prevalence by years of data contributed to the GRPD. RESULTS: In males the point estimate of the prevalence of SLE increased from 7.5/100,000 (CI(95) 6.3, 8.8) to 10.1/100,000 (CI(95) 7.8, 12.2) but this rise was not statistically significant. However, prevalence appeared to increase significantly amongst females from 42.6/100,000 (CI(95) 39.6, 45.6) in 1992 to 70.8/100,000 (CI(95) 65.1, 76.6) in 1998. This increase was mainly amongst women aged 50-79 and in those contributing more than 5 years of data and could not be explained by increasing incidence of SLE or decreasing mortality during the study period. CONCLUSIONS: We found an increasing prevalence of SLE that could not be explained by increasing incidence or decreasing mortality. This is almost certainly an artefact caused by the increased likelihood of detecting or confirming cases of chronic relapsing remitting diseases with increasing time contributed to the GPRD. PMID- 16700094 TI - Unconscious mind can help with complex decisions. PMID- 16700095 TI - By the way, doctor. In your December 2005 article on lung cancer, you mention "certain other airborne substances" as risk factors. What about mold spores? PMID- 16700097 TI - Outbreak news. Avian influenza, Indonesia--update. PMID- 16700098 TI - Cornwall goes public on HIV outbreak. PMID- 16700096 TI - Bergmann glia utilize active caspase-3 for differentiation. AB - Recently, functions associated with caspase have been modified from their well established role in apoptosis. Although caspases are still regarded as mediators of apoptosis, some of the pro-apoptotic caspases, namely caspase-8, -14 and -3 also regulate differentiation in certain cell types, namely myelomonocytic cells, osteoblasts, skeletal muscle cells, keratinocytes, and T lymphocytes. In the central nervous system, non-apoptotic active caspase-3 expression has been located in proliferating and differentiating neuronal cells of the ventricular zone and external granular layer of the developing cerebellar cortex. We previously demonstrated that active caspase-3 expression was not limited to neuronal cells but also was located in the Bergmann glia of the postnatal cerebellum. In that study, active caspase-3 immunolabeling did not markedly colocalize with Ki67, a proliferation marker, but was present in differentiating Bergmann glia that expressed brain lipid binding protein (BLBP) and thus, by its localization, suggested a role in the differentiation of Bergmann glia. The current study addresses the function of caspase-3 in Bergmann glia development by utilizing a Bergmann glial culture preparation. Inhibition of caspase-3 activity by the peptide inhibitor, DMQD-FMK, increased the number of proliferating precursor glial cells and decreased the number of differentiating Bergmann glia, without significantly altering the non-glial active caspase-3 negative population. The transformation in the developmental state of Bergmann glia occurring after suppression of caspase-3 activity strongly suggests an involvement of this enzyme in promoting differentiation of Bergmann glia. PMID- 16700099 TI - Acne vulgaris in monozygotic twins. PMID- 16700100 TI - Heartburn. How to relieve the discomfort. PMID- 16700101 TI - Health tips. Nighttime leg cramps. PMID- 16700102 TI - Dietary antioxidants influence macular degeneration risk. PMID- 16700103 TI - Pain relievers and alcohol. A potentially risky combination. PMID- 16700104 TI - When serious illness strikes. Helping a friend in need. PMID- 16700105 TI - I've read that there are a lot of antioxidants in brewed tea. Does instant tea also have antioxidants? PMID- 16700106 TI - Cardioprotective effects of grape seed proanthocyanidins, baicalin and wogonin: comparison between acute and chronic treatments. PMID- 16700107 TI - Cleaner air on the fly? PMID- 16700108 TI - Prevention of assisted suicide act. PMID- 16700109 TI - An act concerning family law. PMID- 16700110 TI - An act to provide assistance to foreign countries to combat HIV/AIDS, tuberculosis, and malaria, and for other purposes. United States Leadership Against HIV/AIDS, Tuberculosis, and Malaria Act of 2003. Public Law 108-25. PMID- 16700111 TI - An act relating to genetic information; providing that it is an unlawful employment practice for an employer, labor organization or an employment agency to discriminate against a person based on genetic information; and providing other matters properly relating thereto. PMID- 16700112 TI - Licensed to kill: the "death with dignity" initiative. PMID- 16700113 TI - Physician aid in dying: a proposed law for California. PMID- 16700114 TI - The constitutionality of court imposed contraception as a condition of probation. PMID- 16700115 TI - Silver is still confusing. PMID- 16700116 TI - Winter count: taking stock of abortion rights after Casey and Carhart. PMID- 16700118 TI - Product classification. PMID- 16700117 TI - What Lawrence v. Texas says about the history and future of reproductive rights. PMID- 16700119 TI - Saving Roe is not enough: when religion controls healthcare. PMID- 16700120 TI - Econometric analyses of U.S. abortion policy: a critical review. PMID- 16700121 TI - The battle of the bulge and ostomy care. PMID- 16700122 TI - The abortion debate thirty years later: from choice to coercion. PMID- 16700123 TI - Casey and its impact on abortion regulation. PMID- 16700124 TI - Prevention and treatment of perineal skin breakdown due to incontinence. PMID- 16700125 TI - Lethal experimentation on human beings: Roe's effect on bioethics. PMID- 16700126 TI - Adapting a soft silicone dressing to enhance infant outcomes. PMID- 16700127 TI - Enthusiasm on the front lines: "the best job I ever had!". Interview by Barbara Zeiger. PMID- 16700128 TI - New opportunities for wound care clinicians: taking the leap into industry. PMID- 16700129 TI - Abstracts from the 19th Annual Symposium on Advanced Wound Care and the 16th Medical Research Forum on Wound Repair. April 30, 2006-May 3, 2006. San Antonio, Texas, USA. PMID- 16700130 TI - Environmental health nursing: putting knowledge into practice. PMID- 16700131 TI - Government responds to avian quarantine review. PMID- 16700132 TI - [Guidelines for diagnosis and treatment of nonalcoholic fatty liver diseases]. PMID- 16700133 TI - [Concomitant nodular goiter during IFN and ribavirin treatment of HCV: a case report]. PMID- 16700134 TI - [Guidelines for diagnosis and treatment of alcoholic liver diseases]. PMID- 16700135 TI - Outpatient commitment: the state of empirical research on its outcomes. AB - This article briefly describes the historical conditions in the origin and development of outpatient commitment that framed the discourse on its merits and the empirical studies on its outcomes. It divides those empirical studies into two sets on the basis of the questions addressed and critically reviews them. The review pays particular attention to the latest studies that were able to randomize subjects to experimental and control conditions and that were able to offer enhanced services. Finally, this article presents issues not addressed by the empirical studies on outpatient commitment but that need to be addressed in order to understand the choice of using the law to force persons with mental illness to comply with treatment and receive services in the community. PMID- 16700136 TI - Outpatient civil commitment: a dangerous charade or a component of a comprehensive institution of civil commitment? AB - This article examines three criticisms frequently directed toward preventive commitment as one form of outpatient commitment. These criticisms contend that preventive commitment (a) abandons the dangerousness criteria for civil commitmnet, (b) promotes unwarranted inpatient commitment of those who do not meet civil commitment criteria, and (c) undermines important individual liberties by diluting the right to refuse treatment. Understanding and evaluating these criticisms requires analysis of the intersection among empirical, conceptual, and justificatory claims. According to the analysis presented here, advocates of preventive commitment can defend a legitimate role for preventive commitment. This analysis applies to preventive commitment as a dispositional alternative within a comprehensive institution of civil commitment involving distinct parens patriae and police power components. PMID- 16700137 TI - Endorsement of personal benefit of outpatient commitment among persons with severe mental illness. AB - This study examines whether individuals who experienced involuntary outpatient commitment (OPC) attribute benefit to this intervention. It was found that the majority of experimental subjects who underwent a period of OPC did not personally endorse OPC's benefits at the end of the study, either because they did not think it improved treatment adherence or because they rejected their own need for continued treatment. However, at the end of the study, a positive appraisal of benefit was roughly twice as likely among subjects who actually experienced positive treatment outcomes. These data provide little support for acceptance and "gratitude" as a rationale to support decision making about OPC continuation. Rather, clinicians need to rely on other clinical and empirical data for such decision making. PMID- 16700138 TI - Involuntary outpatient commitment. AB - This article offers a novel approach to outpatient commitment (OPC). After distinguishing 4 varieties of OPC, the article shows how 3 are easy to justify, whereas "preventive outpatient commitment" (POC) requires more careful scrutiny. The article argues that, as a general matter, POC is not justified, except for on a "one free shot" basis. The hope is that patients will come to appreciate the benefits of treatment in the community and will become voluntarily compliant; after one free shot, they are in a much better position to decide. The consequences of noncompliance are also explored. PMID- 16700139 TI - Outpatient commitment: a therapeutic jurisprudence analysis. AB - This article analyzes the legal and therapeutic jurisprudence considerations raised by outpatient commitment. Although older forms of outpatient commitment have both legal and therapeutic advantages, preventive outpatient commitment raises serious legal problems and potential antitherapeutic consequences that may outweigh its claimed therapeutic value. As a result, alternatives are proposed, including wider availability of community treatment and outreach and case management services, assertive community treatment, police and mental health court diversion programs, and creative uses of advanced directive instruments and behavioral contracting. Proposals also are made for how preventive outpatient commitment can be applied more therapeutically, including hearings that accord patients a sense of procedural justice and techniques designed to motivate individuals facing such hearings to agree to accept treatment voluntarily. PMID- 16700140 TI - Does involuntary outpatient commitment lead to more intensive treatment? AB - Studies of involuntary outpatient commitment (OPC) among persons with severe mental illness have concluded that OPC is only effective in improving treatment outcomes when it is sustained for 6 months or longer and is combined with frequent outpatient services. This article explores factors that influence the delivery of outpatient services to subjects in a randomized trial of OPC and finds 2 general patterns. Outpatient visits were more frequent among all subjects with apparent clinical need, regardless of study assignment, and among subjects whose OPC was sustained beyond an initial court order. These results suggest that, in practice, sustained OPC represents a consensual agreement between clinicians and the court to more intensively address the complex needs of persons with severe and persistent mental illness. PMID- 16700141 TI - Therapeutic jurisprudence and outpatient commitment law: Kendra's Law and case study. AB - This article considers the implications of assisted outpatient commitment laws (OPC), with specific focus on New York's "Kendra's Law" through the lens of therapeutic jurisprudence (TJ). In this article, the author offers perspectives on the relationship between involuntary civil commitment, outpatient commitment, and the concept of the "least restrictive alternative"; considers pertinent empirical research, and looks at OPC's controversial relationship to forced drugging. Here, the civil libertarian critique is briefly considered, as well as the MacArthur Research Network research. Finally, the author looks closely at Kendra's Law, providing a brief overview of the law itself, and identifying some "pressure points" and pivotal issues, and considers the TJ implications of Kendra's Law, to determine how it "fits" into the public's "take" on all of mental disability law. PMID- 16700142 TI - Exposing the myths surrounding preventive outpatient commitment for individuals with chronic mental illness. AB - Using New York's "Kendra's Law" as an illustrative vehicle, this article addresses the principal criticisms lodged by opponents of preventive outpatient commitment. The authors argue that preventive outpatient commitment is a useful adjunct to conditional release or placement in the least restrictive alternative that has neither produced revolutionary change in psychiatric commitment standards nor will be used inappropriately to assert governmental control over mentally ill citizens. The authors contend additionally that preventive outpatient commitment does not violate federal constitutional norms or represent bad policy making. The authors acknowledge, however, that the coercion inherent in outpatient commitment schemes may produce certain undesirable side effects. Thus, they explore an alternative approach currently in development in Australia that promotes community-based treatment for chronically mentally ill persons without judicial intervention. PMID- 16700143 TI - [9th National Conference on DNA diagnostics with international participation. Prague, 8-9 December 2005]. PMID- 16700144 TI - [Blood screening of benzodiazepines and tricyclic antidepressants. Results of 5 years of interhospital controls]. AB - Pro bio Qual Association of Lyon have proposed a control which include the control for detecting benzodiazepines (BZD) and tricyclic antidepressants (ADT) since 2000. With this control, we have evaluated the specificity and the sensitivity of techniques used. We have tested the maprotiline reactivity too (tetracyclic antidepressant). We report results achieved with different immunoassay methods and their performances: specificity and sensitivity. The number of laboratories which realize these analyses is constant: these laboratories are hospital laboratories. Two methods are most common used: the fluorescence polarization immunoassay (FPIA) and the enzyme multiplied immunoassay test (EMIT). The evolution of these techniques does square with the evolution of chemistry analysers used in hospital laboratories. For the BZD, the specificity is good. For the ADT, carbamazepine at low concentration (5 mg/L) gives a positive result with FPIA Abbott assay; carbamazepine at high concentration (25 mg/L) gives a negative result with EMIT assay; phenothiazines give a positive response. For the BZD, the analyser Integra gives best results of sensitivity. Results of sensitivity obtained with the kit EMIT DAU are better than results obtained with the kit EMIT Serum. For the ADT, results of sensitivity obtained with FPIA Abbott assay seem better; the adaptation of EMIT assay on Integra analyser gives less good results. The reactivity for the BZD is very different: we can ignore a severe intoxication with alprazolam or lorazepam (response << cut off) but we can give a positive result for a therapeutic concentration of diazepam for example. With ADT, we haven't the same problem. But the reactivity for nortriptyline is less good than the reactivity for amitriptyline. So we should use a "cut off" concentration which corresponds to the best sensitivity and the best specificity. PMID- 16700145 TI - [Phalloidin syndrome: role of Elisa-based assay for the detection of alpha- and gamma-amanitins in urine. Preliminary results]. AB - After consumption of mushrooms containing amatoxins (Amanita, Lepiota, and Galerina species), symptoms usually develop after a long delay (>6 h). Initial symptoms start as severe gastroenteritis, progressing to liver failure and possibly death as a result of hepatic coma. Since the survival rate of poisoned patients is claimed to depend on the time of beginning of efficient treatment, fast and reliable assays for amatoxins in biological fluids are essential. Described analytical methods for amatoxins include high performance liquid chromatography and radioimmunoassay (RIA). Recently, a new enzyme-linked immunosorbent assay (Buhlmann Amanitin ELISA kit) has been introduced as an alternative method to RIA. This ELISA-based assay offers several advantages: no complex extraction procedure is required (vs. HPLC) and no safety precautions concerning radioactivity have to be taken (vs. RIA). From August 2004 to October 2005, a pilot study was performed to test the practicability and the clinical utility of this method in emergency situations. RESULTS: ten urines, 9 serums and 1 faeces from 10 patients suffering from acute gastroenteritis after mushroom ingestions (7 contaminated meals) were analyzed. Definitive diagnosis of amatoxin poisoning was made in 4 cases (3 contaminated meals) on the basis of the anamnesis, laboratory results, and clinical course. A patient developed a severe amatoxin poisoning with urinary amanitins level < 1.5 microg/L (urines were collected more than 72 h after mushroom ingestion). Two patients were paucisymptomatic with urinary amanitins levels >10 microg/L (urines were collected before the 36th hour). CONCLUSION: Urine is the sample of choice for the determination of amatoxins. The most critical factor to invalidate the usefulness of this analysis is time. After 36 h, the sensitivity is unreliable. PMID- 16700146 TI - [Heroin substitution treatments. Analysis of current solutions, trends, and perspectives]. AB - Drugs used for treatment of narcotic addicts (buprenorphin at high concentration or methadone) have different advantages and disadvantages. The author has tried to insist on what can be complementary between both treatments. Two new formulations of buprenorphin at high concentration (Suboxone and Subutex NF) are described and proposed to replace Subutex. PMID- 16700147 TI - [The Lafarge affair: a judgment in which toxicology didn't win fame!]. AB - The famous Lafarge affair which took place in Tulle (Correze, France) in 1840, has never ceased to interest the general public because some doubts have remained even after the pronunciation of the judgment. Another hypothesis than arsenicum poisoning has been formulated in 1980 by Professor P. Lepine who believes in a paratyphoid fever occuring after food poisoning. This assumption and the discovery of exceptional family documents in 2004 by Edouard de Lamaze, a distant cousin of Emma Pontier, the confidante of Marie Lafarge, sheds a new light on this mysterious affair. PMID- 16700148 TI - [Fatal methadone poisoning: observations at the CHU of Liege]. AB - All methadone-associated deaths from October 2002 to April 2005 are analysed. A regular increase in these fatal intoxications has been recorded, in Belgium as in other countries, due, in particular, to an intensified prescription of this product, of which illicit use as street dope becomes problematic. Over the 30 months period covered by the study, 26 deaths related to methadone were listed, of which 3 occurred in accidental circumstances. In the other 23 cases, methadone can be found, yet always associated with psychotropic substances, mainly benzodiazepines (18 cases), narcotics (15 cases) and finally alcohol (5 cases). Based upon the nature of the products combined with methadone, records have been divided in two groups: In the first group (17 observations), xenobiotics at (infra)therapeutic levels are detected. In the other group (6 observations), xenobiotics at high and toxic levels are detected. Blood methadone concentrations are not so different between the 2 groups of individuals since the median values and the extreme values are worth respectively 308, and 110-11300 microg/L, for the first group and 776 and 93-2080 microg/L for the second group. There is thus an important overlap between the therapeutic blood methadone concentrations (150 400 microg/L) and blood concentrations observed in fatalities.Thus, it is necessary that all information and post-mortem results must be examined in a critical way to identify and justify cause of the death. PMID- 16700150 TI - Analysis of paraphenylenediamine. AB - The toxic Paraphenylenediamine is characterized by infrared spectrophotometry and nuclear magnetic resonance spectroscopy, whereas, high performance liquid chromatography with a refractive index detector was used to determine its purity in the suspect samples using external standardisation. An analytical method for determination of lower traces of paraphenylenediamine in post-mortem biological fluids was developed. This procedure involves deproteneization or hydrolysis followed by liquid-liquid extraction and derivatization with trifluoroacetic anhydride. 1 microL of the extract was then analysed by gas chromatography/iontrap mass spectrometry. Benzidine used as the internal standard for quantification and the extraction recovery test was evaluated to 85%. This method was validated in cases with paraphenylenediamine poisoning. PMID- 16700149 TI - [Accidental ingestion of methadone and buprenorphine by children. A case review of the Lille Poison Centre between 1995 and 2005]. AB - We have performed a retrospective study (1995-2005) on 218 accidental intoxications in children less than 15 years old, linked to drugs used in the management of opiate withdrawal (cases registered at the Lille poison centre) This study shows a peak of frequency in children less than 3 years old, with a predominance of boys. Poisonings with buprenorphine are more frequent but ingestions of methadone are often more severe (p = 0.004). PMID- 16700151 TI - Generic and brand-name drugs. Are different criteria sufficiently taken into account before granting market authorisation? PMID- 16700152 TI - [Case of death occurred after an injection of aetoxisclerol. The responsibility of the product should be discussed]. AB - A 35-year-old woman has suffered from faintness with cardiovascular failure at the end of a sclerotherapy session for varix management. The injected product was Aeloxisclerol (DCI lauromacrogal 400). The death came up very quickly. The autopsy didn't reveal any traumatic lesion. The anaphylactic choc could be ruled out. Macroscopically, the heart showed a right ventricular dilatation. The toxicological analysis didn't reveal any medicinal substances. Histology showed the presence of lipid degeneration of all the right-ventricle wall The discussion is carried out on the role played by this previous health-state on the onset of death, and also on the incidence of sclerotherapy using this product. and the onset of death. We are carrying out this discussion by taking in mind the nature of the product injected and the information available in the literature concerning the lipid degeneration of cardiac muscle. Quickly after this case, the AFSSAPS (The French Agency for Sanitary Security of Health Products) has emitted an written alert which correlates with the requirements of principles of precaution although it is not founded on the totality of investigation results done for this medicolegal case. PMID- 16700153 TI - [Harmonisation of validation strategies and estimation of the associated uncertainty in the framework of assay laboratoires accreditation]. AB - In order to achieve the harmonization of validation strategies, the interpretation of several validation criteria is proposed. Furthermore, a simple and visual decision tool to assess the validity of an analytical procedure is described: the accuracy profile based on the estimation of the total error of the measurements. This profile build with beta-expectation tolerance intervals can also compute with efficiency the uncertainty related to the results of a laboratory, which is an essential parameter for the accreditation of laboratories under ISO 17025. PMID- 16700154 TI - [The transfer of an automated method for the determination of noradrenaline in human urine: using total error as decision criterion]. AB - Two new statistical approaches based on the total error of measurements were applied to the transfer of an automated method for the determination of noradrenalin in human urine by LC-ECD coupled on-line to SPE. They showed that the receiving laboratory was mastering the analytical procedure allowing it to use the method in routine. Furthermore the risk based approach gave guarantee that the risk to have future measurements out of specification in the receiving laboratory was smaller than 5%, therefore being a risk management tool. PMID- 16700155 TI - [Comparison of three approaches for uncertainty estimation]. AB - Three different approaches for the estimation of uncertainty measurements using the same analytical method were compared, namely validation, robustness and inter laboratory studies. The uncertainty obtained with the robustness study! predicted well the uncertainty of the inter-laboratory study. On the other hand, the uncertainty estimation obtained with the validation study is lower than those obtained with the two other approaches but is still acceptable as long as the analytical method will be used in a single laboratory. PMID- 16700156 TI - [Assessment of occupational exposure to ortho-toluidine using gas chromatography mass spectrometry]. AB - Ortho-toluidine is a carcinogen aromatic amine. It is in part eliminated as unchanged form and its urine determination allows biologic monitoriing of occupational exposure. We propose a new method simple and fast in gas chromatography mass spectrometry. In the Nord-Pas-de-Calais region, a company initiated destruction and depollution of an old industrialsite.The GS-MS method permits exposition evaluation of workers employed in demolition of a liquid SO2 plant polluted with ortho-toluidine. This plant has been stopped twenty years ago. These results are compared with results of workers without any exposure in the same company. A 5 mL urine sample spiked with internal standard (ortho toluidine D9) is extracted with hexane. Derivatisation is achieved with anhydrous pentafluoropropionic acid during 30 min at 60 degrees C. Chromatographic separation is performed on a BPX5MS column (25 m x 0.25 mm, 0.25 microm; SGE). Initial column temperature (60 degrees C) is hold 3 min then is raised to 300 degrees C at 25 degrees C/min. Detection is performed with mass spectrometry with negative chemical ionisation with methane. Acquisition is performed in single ion monitoring. Identification ions are 233 ion (m/z) and 213 ion (m/z) with 233 (m/z) used for quantification. Linearity of the method is verified between 0.1 and 100 microg/L. The limit of detection is 0.02 micro/L. Repeatability and intermediate fidelity are satisfactory (CV < 9%). For unexposed workers, urinary concentrations of ortho-toluidine ranged between 0.17 microg/L and 2.46 microg/g creatinine. Urinary concentrations for exposed workers ranged between 26.14 and 462.00 microg/g creatinine and after new action of protection between 2.35 et 20.11 microg/g creatinine. This new GC-MS method is specific and sensitive and allows for urinary determination of ortho-toluidine. Results showed that this method is adapted for biomonitoring as much for unexposed workers to this aromatic amine as for exposed workers. PMID- 16700157 TI - [Fatal poisoning caused by the ingestion of a concentrated solution of 2,4-D and MCPP]. AB - We report a clinical case of lethal ingestion of an herbicide containing 100 g/L of 2,4-D and 400 g/L de MCPP. The patient shows quickly disturbances of consciousness and cardiac arythmy, a severe metabolic acidosis and an hyperkalemia. The digestive endoscopy at day 4 after ingestion shows an haemorrhagic mucous membrane at oesophagus and stomach level with numerous aulcerations. The bronchial endoscopy shows an inflammatory mucous membrane covered with haemorrhagic liquid. At day 6, appearance of a toxic medullar aplasia. The patient dies at day 7. the autopsy shows haemorrhagic digestive lesions, a bilateral pneumopathy, lungs oedema, an ascite but no cerebral oedema. PMID- 16700158 TI - [Treatment of pesticides determination requests in biological samples: some problems frequently encountered ]. AB - Currently, the determination of pesticides in human fluids is complex. Our scientific knowledge on pesticide intoxication is fragmentary. This leads to numerous difficulties regarding the diagnosis, the analysis request, the choice of the analytical technique, the interpretation of the results. Consequently the waiting time to provide a result is not compatible with the initiation of a treatment in the intoxicated patients. Very few information about Human intoxication with pesticides is available from the literature. In this context, we tried to acquire our own experience. We chose to deal with all pesticides intoxication suspicions received, in collaboration with clinicians. We are presenting the main problems we encounter. PMID- 16700159 TI - Medicine based on systematic research, eminence based medicine or common sense medicine--what would you prefer? PMID- 16700160 TI - The Research Board of EDTNA/ERCA: research activities and deliverables. AB - The Research Board (RB) of EDTNA/ERCA was formed ten years ago to provide opportunities for our members to participate in collaborative research projects at a European level. It provides educational support in research methodology via workshops at conference and in articles published in the Journal. The group has completed a total of seven collaborative research projects in diverse areas of renal care and has recently developed an ambitious new project, the European Practice Database (EPD). The EPD project aims to capture the current practice of renal care professionals in European centres on a three-year repeating cycle. The results of projects are presented at conference and at international scientific meetings by invitation and abstract submission. Recommendations, guidelines or educational material are produced collaboratively with the renal multi disciplinary community. PMID- 16700161 TI - Comparison of renal care practice in Europe: centre and patient characteristics. AB - The European Practice Database (EPD) contains data from 8 countries including 276 centres and 21,861 dialysis patients. Comparing patients and centre characteristics, remarkable similarities and pronounced differences in renal practice between different European countries and between centres within each country were found. PMID- 16700162 TI - Development of indicators to measure European variation of nursing activities. AB - This paper describes a study to capture the key roles and activities of nephrology nurses across different countries in Europe. The concept of the study and the need to clarify the activities of the nephrology nurse arose as part of a larger study to develop the European Practice Database (EPD) (1). The Research Board (EDTNA/ERCA) needed to identify key questions that would detect significant differences in the role and responsibilities of nephrology nurses in different countries and monitor the evolution over time of nephrology nursing practice in Europe. It was therefore appropriate to devise a separate small study to generate evidence based questions for the EPD and confirm the reliability and usefulness of the information captured. PMID- 16700164 TI - Technical aspects of haemodialysis treatment: comparative results of Scotland and Belgium. AB - This paper reports the findings from the European Practice Database project providing an overview of the technical aspects of haemodialysis treatment, comparing practice in Scotland and Belgium. Findings identified that in Scotland almost all centres (95%) employed renal technicians compared to 67% of centres in Belgium. There was also a higher level of technical duties performed by technicians in Scotland compared with Belgium, and marked differences in the disinfection protocols of dialysis machines between the two countries. The study highlights the need for further investigation to discover why technical practice varies somuch between the two countries. PMID- 16700163 TI - European practice in haemodialysis: results of the EPD. AB - The European Practice Database (EPD) project, developed by the EDTNA/ERCA Research Board, collects data on renal practice at centre level in different European countries. Results presented in this paper focus on the European Practice in Haemodialysis centres from 8 European countries or regions following data collection from 2002 to 2004. These results will enable international comparison in practice and will stimulate further research and the development of new practice recommendations. PMID- 16700165 TI - European Practice Database in Belgium. AB - After the successful pilot EPD Project in 2002-2003, we decided to perform the EPD project in four additional countries: Belgium, Greece, Norway and Scotland. Before starting the EPD Project in Belgium, we installed a Belgium National EPD Board in order to support the practical organisation of this project. National coordinators were Luc Picavet (Chair) and Sylviane Boulle (Wallonia). The members of the Belgium National EPD Board were Jean-Yves De Vos (President of National Association, ORPADT Flanders), Hichem Marzougui (Link member, Association of Dialysis Technicians) and Luc Vonckx (President of Association of Dialysis Technicians). PMID- 16700166 TI - Transplant practice in Europe: selection of patients. AB - This paper reports the findings from the European Practice Database project (1) providing an overview of transplant practice in Europe and patient selection. From the 276 centres surveyed 25 recorded performing patient transplants across eight different countries. The overall figures indicate that the waiting list for transplants is made up of 38% women and 9% of patients above the age of 65 years. All countries offered pre-transplant counselling and screening and post transplant follow-up, but the extent to which these activities occurred varied. The waiting/transplant ratio ranged from one in Norway to eight in Slovakia. Differences still exist as a result of country specific policies and legislation regarding transplantation. PMID- 16700167 TI - Infection control practice across Europe: results of the EPD. AB - The European Practice Database (EPD) project, developed by the EDTNA/ERCA Research Board, collects data on renal practice at centre level in different European countries. Results presented in this paper focus on infection control practice in haemodialysis centres from 8 different European countries or regions following data collection from 2002 up to 2004. The prevalence of hepatitis B (HBV), hepatitis C (HCV), human immune deficiency (HIV) and methicillin-resistant staphylococcus Aureus (MRSA) was studied as well as the use of screening and preventive actions. These results will enable international comparison in practice and will stimulate further research and the development of new practice recommendations. PMID- 16700168 TI - European study on epidemiology and management of hepatitis C virus (HCV) infection in the haemodialysis population. Part 3: prevalence and incidence. AB - An analysis of the literature showed a high prevalence of HCV in the European dialysis population in the nineties. The prevalence was similar in most countries in northern Europe, but infection was more common in France, Italy, Spain, Portugal and Greece (1) and in Eastern European countries (2). The reported prevalence of anti-HCV-positive patients in the EDTA registry was 21% in 1992 and 18% in 1993 (3) ranging from 1% in Finland to 42% in Egypt (4). The incidence of HCV, in new patients starting renal replacement therapy, ranged from 3% to 7% (5,6) and reported seroconversion rates during dialysis treatment varied between 1% (7) and 16% (8) per year. PMID- 16700170 TI - The organisation of paediatric renal care in different European countries: results of the PAC project. AB - The Paediatric Access Care (PAC) project, organised by the Research Board of EDTNA/ERCA, aimed to study the organisation of paediatric renal care in Europe and to investigate the practice of access care for both haemodialysis (HD) and peritoneal dialysis (PD) paediatric patients. This paper reports on the organisation of paediatric renal care. The majority of paediatric renal care units were located in specific paediatric units of university hospitals. Most of the centres had offered HD, PD and transplantation (Tx) for more than 20 years. Half of nursing staff had qualifications in paediatric and renal nursing. Most of the centres offered an extended multidisciplinary team approach with the family actively involved in the care of the patient. PD and HD were equally used. Automatic Peritoneal Dialysis (APD) was offered as the standard PD treatment in 2 out of 3 centres. The HD schedule mostly utilised was 3 x 4 hours a week. Half of the patients were on the Tx waiting list and one third of registered patients were transplanted in 2004. PMID- 16700169 TI - Sero-conversion of HCV negative patients: a European study on the epidemiology and management of HCV haemodialysis patients. AB - This paper reports part of the findings from a larger study reported earlier, the European study on epidemiology and the management of HCV in the haemodialysis population (1). Centres recruited to the larger study were monitored for a further one year observation period to measure and generate a deeper understanding of HCV sero-conversion. From 4724 patients who were studied at the baseline, in 68 centres, only 13 patients were found to have sero-converted. These sero-conversions occurred in 7 hospitals within 5 different countries. Possible routes of transmission and risk factors are described with respect to the individual centres and good practice recommendations based on current evidence presented. PMID- 16700171 TI - Paediatric access care in Europe: results of the Paediatric Access Care (PAC) project. AB - Part two of the Paediatric Access Care (PAC) project, a research project of EDTNA/ERCA, investigated PAC in HD and PD patients including the policy for the creation and maintenance of access, and the registration of access related complications that occurred during the registration year of 2004. Data were collected from 39 centres of 13 European countries and included 379 paediatric patients. Fatal complications, resulting in terminating the use of the access, were noted in 59 HD and 22 PD patients. Paediatric access care varied considerably between European centres and in many areas consensus or best practice evidence is still lacking. There is a need for recommendations for the paediatric renal nurse, handling access care in the paediatric renal population. PMID- 16700172 TI - A new Internet resource for chronic kidney disease patients. AB - This paper focuses on the development of a portal in the World Wide Web (WWW), which captures and locates quality information for patients with chronic kidney disease (CKD). It examines the problems patients face when accessing and understanding information gleaned from Web sites and describes an idea from a Research Board Member to facilitate patient access to quality information. The idea germinated into the development of a patient specific Web site, providing one stop access and links to appropriate CKD information, assessed by patients and health professionals. Collaboration between the EDTNA/ERCA Research Board and CEAPIR the European Federation of Kidney Patients has enhanced the project. PMID- 16700173 TI - Association between clinical characteristics, computed tomography characteristics, and histologic diagnosis for cats with sinonasal disease. AB - The purpose of this retrospective study was to determine the association between clinical characteristics, computed tomography (CT) characteristics, and histologic diagnosis in 43 cats with sinonasal disease. All cats were evaluated with CT and nasopharyngeal endoscopic examination, with histologic diagnosis based on nasal biopsy. Fifteen cats were diagnosed with sinonasal neoplasia and 28 cats were diagnosed with rhinitis. Clinical characteristics determined to be significantly associated with neoplasia were unilateral ocular discharge (odds ratio [OR] 9.6) and the presence of a nasopharyngeal mass during endoscopic examination (OR 18.9). CT characteristics found to be significantly associated with neoplasia included: unilateral lysis of ethmoturbinates (OR 11.0), unilateral lysis of the dorsal (OR 8.3) and lateral maxilla (OR 6.9), lysis of the vomer bone (OR 6.7) and ventral maxilla (OR 28.8), and bilateral lysis of the orbital lamina (OR 4.1); unilateral abnormal soft tissue/fluid within the sphenoid sinus (OR 15.3), frontal sinus (OR 10.4), and/or and retrobulbar space (OR 12.2). Lysis of the maxillary turbinates, nasal septum, nasal bone, palatine bone, and cribriform plate were not significantly associated with sinonasal neoplasia. PMID- 16700174 TI - Comparison of conventional spin-echo and fast spin-echo magnetic resonance imaging in the canine brain. AB - T2-weighted fast spin echo and conventional spin echo are two magnetic resonance (MR) pulse sequences used to image the brain. Given the same scan parameters the resolution of fast spin-echo images will be inferior to that of conventional spin echo images. However, fast spin-echo images can be acquired in a shorter time allowing scan parameters to be optimized for increased resolution without increasing the time to an unacceptable level. MR imaging of the brain of 54 dogs, suspected of having parenchymal brain abnormalities was performed using a 1.5 T scanner. Acquisition time ranged from 4 min 24 s to 7 min 16 s (average = 5 min 15 s) for fast spin-echo scans and from 6 min 32 s to 11 min 26s (average = 7 min 55s) for conventional spin-echo scans. All reviewers consistently rated the resolution of fast spin-echo images higher than the conventional spin-echo images (P = 0.000). The potential disadvantages of fast spin-echo acquisitions (motion artifacts, blurring, and increased hyperintensity of fat) did not affect the resolution of the images. Fast spin echo offers increased resolution in a comparable time to conventional spin echo by increased number of excitations and finer matrix size, thus improving the signal-to-noise ratio and spatial resolution, respectively. PMID- 16700175 TI - Value of a single-shot turbo spin-echo pulse sequence for assessing the architecture of the subarachnoid space and the constitutive nature of cerebrospinal fluid. AB - Three case history reports are presented to illustrate the value of the single shot turbo spin-echo pulse sequence for assessment of the subarachnoid space. The use of the single-shot turbo spin-echo pulse sequence, which is a heavily T2 weighted sequence, allows for a rapid, noninvasive evaluation of the subarachnoid space by using the high signal from cerebrospinal fluid. This sequence can be completed in seconds rather than the several minutes required for a T2-fast spin echo sequence. Unlike the standard T2-fast spin-echo sequence, a single-shot turbo spin-echo pulse sequence also provides qualitative information about the protein and the cellular content of the cerebrospinal fluid, such as in patients with inflammatory debris or hemorrhage in the cerebrospinal fluid. Although the resolution of the single-shot turbo spin-echo pulse sequence images is relatively poor compared with more conventional sequences, the qualitative information about the subarachnoid space and cerebrospinal fluid and the rapid acquisition time, make it a useful sequence to include in standard protocols of spinal magnetic resonance imaging. PMID- 16700176 TI - Magnetic resonance imaging and pathologic findings associated with necrotizing encephalitis in two Yorkshire terriers. AB - Two young adult Yorkshire terriers had neurologic signs consistent with forebrain and brainstem involvement or forebrain involvement alone. On magnetic resonance imaging studies there were asymmetric bilateral lesions mainly in the cerebral cortex, and in the diencephalon. These areas were hyperintense on T2-weighted and FLAIR images, but hypointense or isointense on T1-weighted images. Lesions had a varying degree of contrast enhancement. Areas which were isointense on T1 weighted images had no contrast enhancement or only foci of contrast enhancement. Lesions with hypointensity in T1-weighted images had no enhancement or more frequently ring-like enhancement around the lesion. Necrotizing encephalitis was confirmed pathohistologically in both dogs. The degree of contrast enhancement appeared to be related to the degree of lymphohistiocytic inflammation on histologic examination. PMID- 16700177 TI - Vascular anomaly causing subclavian steal and cervical myelopathy in a dog: diagnosis and endovascular management. AB - A 3-year-old dog with cervical myelopathy resulting from a vascular anomaly is described. Marked intradural-extramedullary spinal cord compression was observed, in association with multilevel ectatic anastomotic radicular arterial branches connecting the left and right vertebral arteries. A nonpatent proximal segment of the right subclavian artery had resulted in compensatory enlargement of the left vertebral artery. Flow within the right vertebral artery was retrograde and fed into the patent distal segment of the right subclavian artery. Multiple imaging techniques including myelography, computed tomography, magnetic resonance imaging, and selective digital angiography were used to document this anomaly. To reduce spinal cord compression the largest collateral vessel was embolized without complication. Good clinical response was observed within 6 weeks and improved clinical neurologic function was maintained at the time of a 12-month re evaluation. PMID- 16700178 TI - Helical computed tomographic angiography of the normal canine pancreas. AB - Helical abdominal computed tomography (CT) was performed in nine normal beagle mix dogs. Following cephalic vein injection of ionic iodinated contrast medium via power injector (rate 5 ml/s) dual-phase CT was performed in all dogs. A delayed scan was performed in five dogs between 5 and 13 min after the contrast medium injection. The median time of appearance of contrast medium in the aorta and gastroduodenal artery was 6.3 and 7 s, post start injection and 12 and 12.2 s in the gastroduodenal and portal vein, resulting in a purely arterial pancreatic time window of 5-6s. Pancreatic veins and parenchyma remained enhanced until the end of the dynamic scan (40s). The pancreatic parenchyma showed heterogeneous arterial and homogenous venous contrast enhancement which was slightly hypoattenuating compared to the liver. Delayed scans provided best delineation of the pancreas from the liver. The common bile duct could be identified ventral and to the right of the portal vein joining the dorsomedial aspect of proximal duodenum. Because of the very short time window and variable onset of pure arterial enhancement careful planning of dual-phase studies with previous dynamic CT is recommended. Dual-phase CT angiography enables assessment of the arterial supply, parenchymal perfusion and venous drainage of the canine pancreas. PMID- 16700179 TI - Congenital scoliosis in a quarter horse filly. AB - A 4-week old Quarter Horse filly was evaluated for abnormal gait and lateral deviation of the cervical spine. Physical examination findings prompted radiographs and computed tomography of the thoracic vertebral column which revealed hypoplasia of several thoracic vertebral bodies and resultant scoliosis of the thoracic vertebral column and deviation of the left and right hemithoraces and associated ribs. Collectively, radiography and computed tomography provided an accurate description of the vertebral malformations resulting in scoliosis in this foal. PMID- 16700180 TI - Laparoscopic ultrasonography in six dogs. AB - The aim of this study was to describe the technique of laparoscopic ultrasonography in six dogs with intra-abdominal disease. Investigation of the abdominal cavity was undertaken initially in each dog by laparoscopy alone. After exploration and visualization of a diseased organ, a 7.5 MHz microconvex ultrasound probe was introduced into the abdomen through a laparoscopic cannula for videoguided ultrasound examination. Laparoscopic ultrasonography allows core biopsy or needle aspiration to be performed by guiding the needle into nonsuperficial lesions, where videoendoscopic guidance alone is not enough. In addition, color Doppler ultrasonography is useful to examine the vascular pattern to minimize or eliminate complications when a biopsy is performed. PMID- 16700181 TI - Relationship of pancreatic duct dilation to age and clinical findings in cats. AB - Dilation of the pancreatic duct has been described as an ultrasonographic feature of pancreatitis in cats. The purpose of this study was to determine normal pancreatic duct width in healthy older cats and assess the significance of pancreatic duct dilation observed in a clinical population. In a prospective study, pancreatic ultrasound was performed in 15 healthy cats (mean age 13 +/- 3 years). Mean pancreatic width of left lobe, body, and right lobe was 0.65 +/- 0.16 cm (0.46-1.03 cm), 0.64 +/- 0.14 cm (0.46-0.9 cm), and 0.43 +/- 0.09 cm (0.3 0.57 cm), respectively. Mean pancreatic duct width was 0.13 +/- 0.04 cm (0.06 0.24 cm), which was significantly larger than previously reported for younger cats (0.08 +/- 0.025 cm) (P < 0.001). One hundred and four of 1445 clinical patients (7.2%) were diagnosed with a dilated pancreatic duct and were reviewed in a retrospective study. Incidence of pancreatic duct dilation was significantly higher in older than in younger cats (2.7% in cats < 1-5 years vs. 18.1% in cats 15 years or older; P < 0.001). Mean pancreatic duct width was 0.23 +/- 0.07 cm (0.14-0.52 cm), and there was a significant correlation between age and pancreatic duct width (P = 0.01). There was also a significant relationship between the mean ratio of pancreatic duct width and pancreatic thickness (n = 98) (0.29 +/- 0.09; 0.09-0.58; P = 0.041). There was no significant difference in age between cats with and without pancreatic disease. There was no association between pancreatic disease and pancreatic duct width or pancreatic duct width/pancreatic thickness ratio. Pancreatic duct width and pancreatic duct width/pancreatic thickness ratio in cats are significantly associated with age. PMID- 16700182 TI - Two-dimensional color tissue Doppler imaging detects myocardial dysfunction before occurrence of hypertrophy in a young Maine Coon cat. AB - A 20-month-old healthy male Maine Coon cat was referred for a cardiovascular evaluation. Physical examination and electrocardiogram were normal. The end diastolic subaortic interventricular septal thickness (6 mm; reference range: < or = 6mm) and the mitral flow late diastolic velocity (0.89 m/s; reference range: 0.2-0.8m/s) were within the upper ranges. However, M-mode echocardiography did not reveal any sign of hypertrophic cardiomyopathy (HCM). Tissue Doppler imaging (TDI) identified a marked left ventricular free wall dysfunction characterized by decreased myocardial velocities in early diastole, increased myocardial velocities in late diastole and the presence of postsystolic contractions both at the base and the apex for the longitudinal motion. One year later, the diagnosis of HCM was confirmed by conventional echocardiography and the cat died suddenly 2 months later. This report demonstrates for the first time in spontaneous HCM the sensitivity of TDI for early diagnosis of myocardial dysfunction and suggests that TDI should form part of the screening techniques for early diagnosis of feline HCM. PMID- 16700183 TI - Power Doppler assessment of the neovascularization during uncomplicated fracture healing of long bones in dogs and cats. AB - The aims of this prospective study were to test the feasibility of assessing neovascularization with power Doppler ultrasonography and to investigate its usefulness to follow fracture healing of long bones in dogs and cats. A total of 51 patients (44 dogs and seven cats) were followed. Fracture types differed from simple to comminuted. Therapy ranged from external coaptation to plate osteosynthesis. Patients were followed with radiography, B-mode real time and power Doppler ultrasonography every 2-4 weeks until the fracture was healed. All fractures healed uneventfully. A semi-quantitative numerical score based on signal intensity, vessel area, and number of Doppler signals was assigned and the mean value was used to compare patients and examinations. Time postoperatively was divided into periods of 10 days. No Doppler signal was present during the first 10 days. The mean of the scores was highest between 11 and 20 days postoperatively and the median of the scores peaked between 21 and 30 days. A gradual decrease was seen thereafter. The mean of the scores was zero at 71-80 days and the median at 51-0 days postoperatively for the grouped results. In all positive power Doppler examinations, signals were present in and close to the callus. In seven patients (five dogs and two cats) signals were also present in the peripheral soft tissues in one of the follow up examinations. The normal healing process of fractured bones can be visualized using power Doppler ultrasonography and follows a distinctive time-dependent pattern. PMID- 16700184 TI - Transesophageal echocardiography of the left ventricular outflow tract, aortic valve and ascending aorta in Boxer dogs with heart murmurs. AB - The study was aimed at evaluating the anatomy of the left ventricular outflow tract, aortic valve, and ascending aorta by means of multiplane transesophageal echocardiography in Boxer dogs with left basilar heart murmurs and at comparing two-dimensional (2D) transthoracic to transesophageal echocardiography for the diagnosis of subaortic stenosis in this breed. Twenty-eight Boxers were included in the study and allocated to four groups according to physical and routine transthoracic 2D and Doppler echocardiography findings: group A--dogs with low grade (I-II/VI) heart murmurs without overt evidence of aortic stenosis (14 dogs); group B--dogs with type 1 subaortic stenosis (seven dogs); group C--dogs with type 2 subaortic stenosis (five dogs); group D--dogs with type 3 subaortic stenosis (two dogs). Anatomic lesions were not discovered by transesophageal echocardiography in dogs belonging to group A. Transesophageal imaging confirmed the type of subaortic stenosis, as graded by transthoracic echocardiography, in diseased animals (groups BCD). Morphologic information obtained by transesophageal echocardiography in Boxer dogs was similar to that obtained by transthoracic echocardiography. PMID- 16700185 TI - Echocardiographic and radiographic diagnosis: complete situs inversus in a cat. PMID- 16700186 TI - Pneumonia: how important are local epidemiology and smoking habits? PMID- 16700187 TI - Determinants of quality of life after lung transplant: an Italian collaborative study. AB - BACKGROUND: With the improvement in survival rates after lung transplantation, concern has arisen about evaluating quality of life (QoL). This multicenter cross sectional study aimed at describing QoL and identifying factors associated with it. METHODS: We assessed QoL in 129 lung transplant recipients from 5 centres in Italy, during scheduled follow-up visits, using the SF-36, GHQ and St George's respiratory questionnaires (SGRQ). RESULTS: The SF-36 elicited impaired QoL in the physical, but not in the mental domains (PCS = 44; MCS = 53). The GHQ identified 29 patients (23%) with psychological discomfort and the SGRQ scores were significantly better than those of patients with chronic respiratory disease. On multivariate analysis, exertional dyspnea was an independent predictor of the PCS (adjusted delta -6.3 (p < 0.001), while osteoporosis (delta = -3.1), BOS (delta = -4.3), acute rejection (delta = -3.9) and heart and lung transplant (delta = +6.4) were only marginally associated. Dyspnea was also related to a GHQ score > 5. CONCLUSIONS: The study identified exertional dyspnea as the main determinant of QoL as measured both by SF36 (PCS) and GHQ. Other objective measures contributed only to the PCS. Thus, the SF-36 (PCS) and GHQ were useful in identifying patients who needed treatment not only for complications but also psychological support and continued physical rehabilitation. PMID- 16700188 TI - Demographics and microbiological profile of pneumonia in United Arab Emirates. AB - BACKGROUND: Pneumonia is a common respiratory disease, which has a mortality rate of approximately 14% worldwide. The management of pneumonia is based on the patient and microbiological profile existing in the community. There is a paucity of data regarding patient demographics and the microbiological profile for pneumonia in the United Arab Emirates (UAE). METHODS: We conducted a retrospective analysis of in-patients with pneumonia in the UAE. Epidemiological, clinical and microbiological data was collected from patients over the age of 16 years admitted to a single university hospital with a diagnosis of pneumonia between the years 1997 and 2002. Patients were categorised as having community acquired (CAP) or hospital acquired pneumonia (HAP). RESULTS: Among the 361 patients, 186 were females and 175 males. The mean age (+/- SEM) of the males was 59 +/- 1.6 years and of the females, 56.9 +/- 1.5 years. The annual hospital admission rate for pneumonia increased from 410 per million population in 1997 to 760 in 2002. The pneumonia was community acquired in 289 (80%) and hospital acquired in 72 (20%). Hospitalisation for CAP showed a significant seasonal variation with peak admission rates in March and April and the nadir in August and September. The overall mortality rate was 13%, and was significantly higher for HAP (24%) than for CAP (10%, p < 0.01). The common microorganisms isolated from sputum culture in CAP were Haemophilus influenzae (18.6%) and Streptococcus pneumoniae (10%). The common pathogen in HAP was Pseudomonas aeruginosa in (50%). 4 out of 17 (23%) H. influenzae isolates were resistant to amoxicillin and 2 of 9 S. pneumoniae isolates (22%) were resistant to penicillin. CONCLUSIONS: The hospitalisation rate for CAP has increased over the years and showed a clear seasonal trend in the UAE. H. influenzae was the most common organism among patients with CAP and P. aeruginosa in HAP. The mortality rate for HAP was significantly higher than for CAP. The drug resistance pattern was similar to reports from centres elsewhere in South East Asia. PMID- 16700189 TI - Differential effect of cigarette smoking on hydrogen peroxide and thiobarbituric acid reactive substances exhaled in patients with community acquired pneumonia. AB - BACKGROUND: This study was designed to investigate the effect of cigarette smoking on hydrogen peroxide (H2O2) and thiobarbituric reactive substances (TBARs) concentrations in exhaled breath condensate (EBC) in patients with community acquired pneumonia (CAP). METHODS: H2O2 and TBARs concentrations in EBC were determined with spectrofluorimetrical assays. RESULTS: Non-smoking CAP patients (n = 24) exhaled 1.4, 1.8 and 1.7 times more H2O2 than the smoking patients with CAP (n = 19) as assessed one (0.73 +/- 0.32 microM v. 0.51 +/- 0.36 microM), three (0.84 +/- 0.31 microM v. 0.47 +/- 0.24 microM) and five (0.66 +/- 0.28 microM v. 0.40 +/- 0.35 microM) days after admission (p < 0.05 in each case). Over 10 days of hospital treatment, mean level of exhaled H2O2 0.45 +/- 0.22 microM in CAP patients with smoking history was decreased if compared with 0.71 +/- 0.19 microM exhaled H2O2 in CAP group (p = 0.005). On the contrary, TBARs concentration evaluated over entire study period was increased in smoking CAP patients (median 0.02 microM, range 0-0.32 microM) compared with non-smoking group (median 0.01 microM, range 0-0.21 microM, p < 0.05). Concurrent, active smoking status was related with the decreased levels of H2O2 exhaled in breath condensate within the course of CAP but it appeared to increase levels of TBARs. CONCLUSIONS: The differential alternations of oxidative parameters in EBC with respect to the smoking status might provide evidence of increased H2O2 decomposition and enhanced generation of reactive species in airways of CAP patients. PMID- 16700191 TI - Allergen-specific immunotherapy: an update on immunological mechanisms of action. AB - Specific immunotherapy (SIT) is the only treatment able to modify the natural history of the allergic subjects. Several aspects of the immunopathological response modified by SIT have been investigated; the first parameter historically studied was the production of allergen-specific antibodies. An increase of allergen-specific IgG4 and a decrease of IgE appear after SIT. A shift from Th2 polarized immune response toward Th1-oriented pattern has been reported after SIT. More recently, a crucial role for a subpopulation of T cells has been evidenced: T regulatory cells (Treg). Allergic patients have a defect of Tregs. SIT is able of inducing a specific Treg response. Sublingual immunotherapy is an alternative route of administration for SIT. Recent evidence shows that SLIT is also able of inducing a Treg response as detected by IL-10 production. PMID- 16700190 TI - Pleural tuberculosis. AB - Pleural effusions in tuberculosis are commonly seen in young adults as an immunological phenomenon occurring soon after primary infection. However, the epidemiology and demographics of tuberculous pleurisy are changing due to the impact of HIV co-infection and the increasing number of pleural effusions seen as part of re-activation disease. Pleural biopsy for histology and culture is the mainstay of diagnosis with closed needle biopsy adequate in the majority of cases. Techniques such as PCR of biopsy specimens and the role of pleural fluid ADA are still being evaluated as a diagnostic aid. Tuberculous empyema is less commonly seen in the western world and the diagnostic yield from pleural fluid here is greater than in "primary" effusions. Treatment with appropriate antituberculous chemotherapy is generally successful though there is currently insufficient evidence to recommend the routine use of corticosteroids in this condition. PMID- 16700192 TI - Dose dependence of efficacy but not of safety in sublingual immunotherapy. AB - Sublingual immunotherapy (SLIT) currently represents, as indicated by meta analysis of its efficacy and safety, a valid option to the generally used traditional subcutaneous immunotherapy (SCIT) for treating respiratory allergy. Regarding efficacy, recent studies demonstrated that, similar to what has already been observed in SCIT as well as in experimental and clinical studies about the magnitudo of allergen exposure, the effectiveness on both clinical symptoms and immunologic changes depends on the amount of allergen administered during treatment. In addition, in vitro studies addressed with the role of dendritic cells, currently considered to be of pivotal importance in orienting toward tolerance the immune response to allergens, showed that the internalisation of allergen molecules, which is followed by tolerogenic presentation to T cells, depends on the amount of allergen. However, such dose dependence is not apparent concerning the safety. In fact, the comparison of studies respectively conducted with high and low allergen doses did not show differences in the rate of systemic reactions, which in any case never had the presentation of anaphylaxis, and instead a significant difference in the rate of local reactions, following the oral and gastrointestinal contact with the allergen extract, in favour of high dose studies. PMID- 16700193 TI - Dose dependence of efficacy and safety of subcutaneous immunotherapy. AB - A number of experimental and clinical evidence has shown that exposure to high amounts of allergen molecules favours the development of tolerance. This is true also for subcutaneous immunotherapy (SCIT), for which a dose dependence of clinical efficacy was clearly demonstrated. The effective doses, measured as microg of major allergens, to be administered during maintenance treatment were established for the main allergens. Regarding pollens, the range of effectiveness corresponds to 25-41 and 13-20 microg of major allergens Phl p 5 and Phl p 6 for grasses, to 10-47 microg of Amb a 1 for ragweed, to 12 microg of Bet v 1 for birch, and to 6.2 microg of Par j 1 for Parietaria. With house dust mites, a maintenance dose of 5-11.5 microg of the major allergen from Dermatophagoides pteronyssinus Der p 1 is associated to clinically relevant effects, and with cat epithelium the clinical success is observed using a dose of 13-15 microg of Fel d 1. Nevertheless, there are adverse reactions facing SCIT, which are related to the amount of injected allergen. In fact, the safety decreases when the administered doses increase. This has led to "optimal doses" being defined which show a good balance between efficacy and safety (corresponding for example to a dose of 7 microg for Der p 1 and of 13 microg for Fel d 1). The dose dependency with respect to both efficacy and safety makes essential to accurately consider the risk/benefit ratio in each patient eligible for SCIT. PMID- 16700194 TI - Practice parameters for sublingual immunotherapy. AB - The efficacy and safety of sublingual immunotherapy (SLIT) are currently supported by clinical trials, meta-analysis and post-marketing surveys. Practice parameters for clinical use of SLIT are proposed here by a panel of Italian specialists, with reference to evidence based criteria. Indications to SLIT include allergic rhinoconjunctivitis, asthma, and isolated conjunctivitis (strength of recommendation: grade A). As to severity of the disease, SLIT is indicated in moderate/severe intermittent rhinitis, persistent rhinitis and mild to moderate asthma (grade D). SLIT may be safely prescribed also in children aged three to five years (grade B), and its use in subjects aged more than 60 years is not prevented when the indications and contraindication are ascertained (grade D). The choice of the allergen to be employed for SLIT should be made in accordance with the combination of clinical history and results of skin prick tests (grade D). Polysensitisation, i.e. the occurrence of multiple positive response does not exclude SLIT, which may be done with the clinically most important allergens (grade D). As to practical administration, co-seasonal, pre co-seasonal, and continuous schedules are available, being the latter recommended for perennial allergens or for pollens with particularly prolonged pollination, such as Parietaria (grade D). For pollens with relatively short pollination, such as grasses and trees (cypress, birch, alder, hazelnut, olive) the pre co-seasonal and perennial schedules are preferred (grade C). The build-up phases suggested by manufacturers can be safely used (grade A), but they can be modified according to the patient's tolerance (grade C). A duration of SLIT of 3-5 years is recommended to ensure a long-lasting clinical effect after the treatment has been terminated (grade C). PMID- 16700195 TI - Lung stem cell update: promise and controversy. AB - Currently, there is great enthusiasm about potential stem cell therapies for intractable diseases. We previously reviewed the topic of stem cells in lung injury and repair, including the role of endogenous, tissue (somatic) stem cells and the contribution of circulating cells to the lung parenchyma. Our purpose here is to provide a concise update in this fast-moving field. New information and ongoing debate focus attention on basic issues in lung stem cell biology and highlight the need for additional studies to establish the feasibility of cell therapies to prevent or treat lung diseases. PMID- 16700196 TI - Euthyroid goitre and sleep apnea. AB - A number of predisposing factors (obesity, nasal obstruction, adenoidal hypertrophy, macroglossia, etc) have been related to obstructive sleep apnea syndrome (OSAS). In addition hypothyroidism and large goitres have been reported to be associated to OSAS, but this association has not been adequately studied. We describe an obese patient with euthyroid goitre associated with OSAS. The patient showed a body mass index (BMI) of 47 and a large neck with a circumference of 60 cm. The flow-volume curve demonstrated an expiratory plateau suggesting an intrathoracic upper airway obstruction. Arterial blood gas analysis results were: pH 7.39; PCO2 54.2 mmHg; P O2 47 mmHg. Nocturnal polisomnography showed an apnea/hypopnea index (AHI) of 31 episodes/hour. Upper airway collapse was overcome by a nasal continuous positive airway pressure (nCPAP) of 14 cmH2O. Weight loss obtained by a hypocaloric diet was not accompanied by any OSAS improvement. After thyroidectomy, a nCPAP of 4 cmH2O was sufficient to prevent upper airway closure. Discontinuation of nCPAP treatment for 4 consecutive nights did not determine worsening of sleep apnea symptoms, nor a worsening of overnight oxymetry. A new polysomnography carried out after 4 nights off nCPAP showed an AHI of 33 episodes/hour. OSAS should be suspected in patients with large goitres. Decisions regarding discontinuation of nCPAP treatment after thyroidectomy should be based on polisomnographic results. PMID- 16700197 TI - Collapse of a new type of self-expanding metallic tracheal stent. AB - Alveolus TB-STS is a new self-expanding, completely polyurethane-covered, metallic stent which has been designed to be successfully used even in the treatment of non-neoplastic airway strictures as it is supposed to be removable. We recently observed the collapse of an Alveolus tracheal stent, causing dyspnea and hemopthysis, in a 63-yr-old female patient with post-intubation tracheal stenosis. Such a complication, which to our knowledge has never been previously reported with metallic stent use, forced us to remove the stent. PMID- 16700198 TI - Fire-eater's pneumonitis. AB - The authors report a case of acute respiratory failure that fulfils the diagnostic criteria for fire-eater's pneumonitis; pathological features, usually described only in experimental works on animals, are also discussed. PMID- 16700199 TI - [The urologist like family practice in aging patients]. PMID- 16700200 TI - [The role of the pelvic physical therapist]. AB - Pelvic physical therapy focuses on the prevention and the treatment of all kinds of functional disorders of the abdominal, pelvic and low back region, like urinary incontinence, that is considered as a high prevalent health problem in women, men, children and the elderly. Physical therapy is often considered as the first-choice treatment, due to its non-invasive character, the results in terms of symptom relief, the possibility of combining physical therapy with other treatments, the low risk of side effects and the moderate to low costs. Important restrictions for success might be motivation and perseverance of patient and therapist and the time needed for physical therapy. The armentum of the pelvic physical therapist contains interventions such as physiotherapeutic diagnostics, education and information of patients, pelvic floor muscle (PFM) training, bladder training (BlT), training with vaginal cones, electrical stimulation, biofeedback, etc. In stress incontinence, to improve the extrinsic closing mechanism of the urethra, physical therapy is aimed on strength improvement and coordination of the peri-urethral and pelvic floor muscles. Especially, PFM training is effective. For detrusor overactivity physical therapy aims to reduce or eliminate involuntary detrusor contractions through reflexinhibition. Here, electrical therapy appears to be an effective intervention. In mixed urinary incontinence the physiotherapeutic diagnostic and therapeutic process focuses on the predominant factors. Radical prostatectomy is the most important cause of incontinence in men. An adequate program of PFM training, after radical prostatectomy, decreases the duration and the extent of incontinence and improves the quality of life. CONCLUSION: physical therapy is in many cases of incontinence an effective treatment option. PMID- 16700201 TI - [Hormone-refractory prostate cancer. Modifications of the therapeutic strategies since chemotherapy proved its usefulness]. AB - Back in the 90's it was difficult to have access to the conclusions of publications on HRPC. Homogeneity was very scarce regarding issues as significant as the definition of HRPC itself, patient selection, or evaluation of the responses to therapy. Consensus has currently been reached on such matters, and it is described in this text. Two works were published in late 2004 showing that docetaxel-based chemotherapy improved metastatic HRPC survival. Until then, the different treatments used could only provide symptomatic relief. But probably not all of the HRPC patients are eligible for primary docetaxel chemotherapy. The current debate focuses on determinating to which patients should chemotherapy be administered and at which time should it start, in order to exclude those patients at risk of experiencing its adverse effects without benefitting from its clinical advantages. Non-metastatic HRPC patients may be candidates to receiving secondary hormone manoeuvres before starting with chemotherapy. We will analyse in this review the changes occurred in the therapeutic strategies ever since chemotherapy showed its value, and we shall also disclose our attitude regarding treatment of these patients in daily practice. PMID- 16700202 TI - [TRUS-guided biopsy: comparison of two anesthetic methods]. AB - PURPOSE: The aim of the present study is to compare two analgesic techniques for ultrasound transrectal biopsy. Oral analgesia vs periprosthetic nerve blockade with 2% mevicaine. PATIENTS AND METHODS: A total of 200 patients were randomized prospectively into 2 groups, namely group I: 100 patients treated with metamizol, oral morphine 30 minutes before the procedure, and group II: 100 patients anesthesied with periprosthetic nerve blockade with 2% mepivacaine. Both groups were treated with bromacepan 3 mg 30 minutes before the biopsy. The first intention was to obtain 10 core TRUS-guided biopsy in all patients underwent. After the procedure, a ten visual analogue pain score (VAS) from 0 = no discomfort to 10 = severe pain was administered to the biopsied patients and a global estimation of pain associated with the procedure was obtained. Test T de Student was used for statistical analysis. RESULTS: There were no significant differences in age, PSA and prostate volume. 3 core TRUS-guided biopsy were obtain in group I (3 +/- 1.3), and 10 in group II (5 +/- 1.2) In the periprosthetic block group (II) 95% of patients referred no pain after the procedure (VAS = 0), 2% middle pain (VAS = 5-6) and 3% strong pain (VAS = 7-8); while patients in group I referred 12.5% no pain, 42.4% middle pain, 20% strong pain. The level of pain reported by this group of patients was significantly different from those reported by patients who performed prostate biopsy with periprosthetic nerve blockade. (p < or = 0.05). There were no significant differences in major complications. CONCLUSIONS: The use of bilateral periprosthetic block with mepivacaine is a very effective and useful technique, well tolerated by the patient, which almost completely abolishes the pain and discomfort associated with the prostate biopsy procedure. And also allows increase the number of cores. PMID- 16700203 TI - [Renal cancer in elderly: clinical and histopathological findings]. AB - OBJECTIVE: to evaluate the clinical and pathological characteristics of the CCR in our series of tumors, analyzing its impact in the group of age greater than 65 years. MATERIAL AND METHODS: 300 patients with renal adenocarcinoma (CCR) were studied. In 252, ploidy pattern of DNA and pathologic characteristics of the surgical pieces were done. According to the criteria of the Spanish Society of Geriatrics, the patients were divided in two groups based on the age, greater and smaller of 65 years. The clinical and pathologic characteristics were compared between both groups. RESULTS: 103 of the 300 patients (33.3%) were included in the geriatric group. We did not found differences between both groups comparing stage, tumoral volume or treatment realized, but found differences in DNA ploidy pattern, recurrences and survey. CONCLUSIONS: The RCC in the elderly has a few clinical and pathological characteristics similar to the rest of patients in our series. Nevertheless differences exist in the average and actuarial survival, which is minor in the patient of more than 65 years, cause the percentage of detected aneuploidies and number of recurrences in not confined tumors in this group of age. PMID- 16700204 TI - [Living donor kidney transplantation: helical CT evaluation of candidates]. AB - OBJECTIVES: Computed tomography (CT) is a diagnostic tool that has won acceptance in the past years for the vascular evaluation of living renal donor (LRD) candidates. We show the findings observed in a series of 102 candidates consecutively evaluated in our centre and present the correlation with the surgical data obtained in those individuals finally operated. METHODS: From May 1995 to June 2005, 102 LRD candidates were evaluated with CT. Eighteen were also studied with digital subtraction angiography (DSA). Fifty-eight donors underwent nephrectomy. Prevalence of arterial and venous variants found on CT exams were analyzed, as well as the correlation with surgical and DSA data when they were obtained. RESULTS: Forty-four percent candidates showed multiple renal arteries at least in one kidney. Seventy-one percent of right kidneys and 76% of left kidneys had a single arterial pedicle. Early branching was observed in 7.8% and 15.7% in right and left kidneys respectively. Venous variants were present in 17.5% and the 13.7% in right and left sides respectively. Sensitivity, specificity and accuracy of CT to detect accessory arteries were 89%, 100% and 97% respectively when it was compared to DSA. Correlation with surgical data raised 93% for the presence of arterial variants. CONCLUSIONS: Vascular variants in renal pedicle showed a high prevalence. Helical CT can effectively achieve all the information required prior to surgery as a sole imaging procedure in LRD candidates. PMID- 16700205 TI - [Diagnosis and management of cystic dysplasia of the seminal vesicle]. AB - INTRODUCTION AND OBJECTIVES: Cystic dysplasia of the seminal vesicle represents a rare congenital urologic anomaly of which elective treatment is controversial. We present our experience in the diagnosis and management of this uncommon pathology. PATIENTS AND METHOD: During the last twelve years we have managed eight patients affected by this pathology. The diagnosis was incidental in the three asymptomatic patients, while the other five consulted because of low urinary tract sympthoms. Ectopic drainage of the ureter in the pathological seminal vesicle was found in three of the patients. Two of these patients presented large pelvic and ureteral ectasy. Other five patients presented renal agenesis. RESULTS: In asymptomatic patients we adopted an expectant attitude, while the other five patients underwent surgical treatment. We performed open surgery in the two cases presenting ureterohydronephrosis, whereas in the other three we performed transurethral incision of the seminal vesicle. We obtained excellent results in all of them, with no evidence of symptomatic pseudodiverticulum. CONCLUSIONS: Transurethral incision of the seminal vesicle cyst is a minimal invasive procedure with low morbidity. We have proved in all of our cases that the retraction of the cyst was practically complete, this has led us into believing that transurethral incision is the elective treatment. Therefore, open surgery would only be indicated in cases of failure of the procedure due to existence of symptomatic pseudodiverticulum, or in patients presenting renoureteral ectasy associated. PMID- 16700206 TI - [Penile prosthesis for the treatment of erectile dysfunction]. AB - The erectile dysfunction is a pathology that, with different degrees of intensity, affects nearly the 20% of the spanish adult men. The treatment is usually performed in stages, reserving the penile prosthesis for when other previous treatments have failed. The aim of this work is to evaluate, according to the state of present knowledge, the effectiveness and security of the penile prosthesis for the treatment of the erectile dysfunction. With this purpose 52 articles were selected, observing a 5 years prosthesis survival of 78-91% and a 3 8% of surgical complications. Mechanical failures and infection percentages were smaller in the semi-rigid prosthesis that in the inflatable ones, with high levels of postoperative satisfaction in patients as well as in their couples, even greater than in other treatments available at the present time. The penile prosthesis implantation must be reserved for the organic erectile dysfunction when previous treatments have failed, evaluating the risk-benefit relation and informing the patient of the results that are hoped to be obtained and of the possible complications that can arise. In view of the great concern of our society with the erectile function and the availability of effective drugs, an increase in the demand of penile prosthesis implantation is predictable in those patients highly motivated, but refractory to the less invasive treatments. PMID- 16700207 TI - [Vesicoureteric reflux endoscopic treatment complications in childhood]. AB - INTRODUCTION: Nowadays, around 1% of children suffer from vesicoureteral reflux (VUR), which represents a risk factor for long-term kidney damage. In the last decade, development in this field has shown that subureteric endoscopic injection of bulking agents for childhood VUR is an acceptable alternative unlike long-term antibiotic prophylaxis and open surgery. OBJECTIVE: To establish the complications of childhood VUR endoscopic treatment. MATERIAL AND METHODS: A retrospective analysis was carried out for every patient treated endoscopically in our department from January 1999 to September 2003, with a total amount of 516 implants of submucous bulking agents. All patients were assessed with an ultrasound as a first diagnostic technique so as to establish the complications associated with it, especially ureteral obstruction and injected agent migration. RESULTS: Out of 516 cases, Macroplastique was used in 455, Deflux in 44 and Polytef in 17. 10 patients (1.9%) (5 girls/ 5 boys), with ages ranging from 15 months to 11,4 years, presented some kind of complications (Macroplastique was used in 8cases and Deflux in 2). Among them, 5 were remarkable due to an ureterohydronefrosis, 2 of which needed a nephrostomy. 3 patients presented an implant migration, 1 epididymitis and 1 bladder edema. CONCLUSIONS: Endoscopic bulking agent injection has become a safe alternative to open surgery in childhood VUR management. PMID- 16700208 TI - [Volume determinations of the whole prostate and of the adenoma by transrectal ultrasound: correlation with surgical specimen]. AB - OBJECTIVE: We evaluated whether preoperative transrecta ultrasound (TRUS) mesaurements of the transition zone (TZ) and total prostate volumen predict real prostatic weight. MATERIAL AND METHODS: We compare estimated TRUS volumes with surgical specimen weight, in surgically treated patients with localized prostate cancer (group A, n = 33) or benign prostatic hyperplasia (group B, n = 37). The volume was calculated by the ellipsoid formula. Both measurements were compared with surgical specimen weight, assuming 1 as specific prostate weight. RESULTS: Group A: mean prostate measured volume was 38.6 cc. (SD 22.7), mean RP specimen weight was 54,2 g (SD 27.2) (p = 0.001). Total estimated prostate volume underestimated prostatectomy specimen weight by 29%. In order to adequate the estimated volume to the specimen weight, we calculated the formula: estimated prostate weight = 0.95 x prostatic measured volume + 17,657 (p = 0.005). Group B: mean TZ measured volume was 62.8 cc. (SD 23.3), mean adenomectomy specimen weight was 79.9 g (SD 45.9) (p = 0.001). TZ estimated volume underestimated adenomectomy specimen weight by 21%. In order to adequate the estimated volume to the specimen weight, we calculated the formula: estimated TZ weight = 1.67 x TZ measured volume - 24,768 (p = 0.04). CONCLUSIONS: We found significative differences between TRUS measured volumes and real weight of surgical specimen. These differences could be corrected by simple formulas that allow to minimize the observed underestimations. PMID- 16700209 TI - [TVT our experience five years and six months later]. AB - OBJECTIVE: We present the outcome of urinary incontinence surgery after TVT more than five year of follow-up. Efficacy was evaluated in terms of complete cure of incontinence, complications, and patient satisfaction. MATERIAL AND METHODS: A sample of 369 female with genuine stress incontinence or mixed incontinence who had a TVT performed in our hospital between 1998-2003 were evaluated. Sample average age was 59.9 years. A total of 326 patients (85%) had genuine stress incontinence and 56 (15%) mixed incontinence. Preoperative evaluation was base on: clinical history and physical examination, stress test, flowmetry and post voided residual. Cistometry was indicated only in patients with previous surgery and mixed incontinence. Severity of incontinence was graded clinically. Primary outcome measure was complete cure of incontinence defined as lack of objective and subjective leakage. In addition a phone survey about patient satisfaction with the surgery was carried out. RESULTS: The average follow up was 35 months (6 67). A total of 317 were complete continent (86%). By contrast, 52 patients (14%) had leakages jet. The novo urgency rate was 7% and a 5.7% of patients were reoperated to cut the mesh because of obstruction. Nevertheless, the reoperation rate was low and only a patient showed a massive haematoma which needed to be evacuated. CONCLUSION: Bursch Technique through suprapubic route has been classically considered the gold standard procedure for stress urinary incontinence for the last decades, having been substituted nowadays by tension free suburethral mesh sling whose most important advantages are: Simplicity of the method, lower cost and higher comfort for the patient due to the fact that we have chaged in-patient surgery program for ambulatory one with similar results in order to sanation. Anyway we will have to check if the preliminary good results and no complications persist in the follow-up. PMID- 16700210 TI - [TVA and TOA. New adjustable mesh for the treatment of female stress incontinence. Preliminaries results]. AB - OBJECTIVES: Evaluation of a new mesh for incontinence (TVA/TOA) which enables the degree of tension applied during surgery to be readjusted at the post-operative stage. PATIENTS AND METHOD: 62 patients treated with the TVA mesh and monitored over a 14-month period (SD 7.8, range 6-38). In 33 patients (53%) some other pelvic prolapse was corrected. Evaluation was carried out by clinical report, examination of bladder full of 250 saline solution, flowmetry and urinary residue. 40 patients filled in 4 quality of life questionnaires (QoL; ICIQ-SF; PGI-S; PGI-I). RESULTS: 42 patients were found to be objectively continent in the post-operative evaluation. Of these, it was necessary to reduce tension in 7 cases (11%) due to urinary obstruction (flow < 10 ml/sec and/or residue). The tension of the mesh was tightened in 20 patients (32%) due to continue with a certain degree of incontinence. All patients were discharged as continent and with no residue. In the last revision, 58 patients (93%) proved to be objectively continent and 4 (6.5%) showed a notable improvement in their incontinence. The Q(MAX) is 19.8 ml/sec (SD 9.8). The mictional urgency had disappeared or improved in 32 of the patients who had this prior to operation (76%) and had appeared in 3 of the patients who didn't (15%). The clinical report showed a high level of consensus with the ICIQ-SF survey (Kappa = 0.89) regarding stress incontinence, diminishing clearly (Kappa= 0.13) when urge incontinence was taken into account. 34 (85%) patients scored over 95 out of 110 in the QoL. 30 (75%) scored less than 6 in ICIQ-SF. 32 (80%) showed a perception of normality and 4 (10%) slight illness in the PGI-S. In the PGI-I 29 (72.5%) were much better and 11 (27.5%) quite a lot better. A relation exists between urgency and dismissed quality of life. CONCLUSION: With the TVA (trans-vaginal adjustable) mesh it is possible to adjust the tension originally applied during surgery at the post-operative stage, so that any defects or excesses can be corrected. PMID- 16700211 TI - [In vitro three-dimensional reconstruction of human bladder mucosa]. AB - OBJECTIVE: The purpose of this study is to apply the in vitro keratinocyte culture techniques and the tissue engineering principles to human urothelium, to reconstruct an in vitro three-dimensional human bladder mucosa, suitable for grafting. MATERIAL AND METHODS: Biopsy specimens of human bladder mucosa were obtained from patients undergoing suprapubic prostatectomy, in vitro cultured and finally, an immunohistochemical study was made. RESULTS: A three-dimensional in vitro tissue was obtained, composed of a bio-artificial submucosa (fibrin gel and fibroblast) where the uroepithelial cells were seeding. We used a biodegradable polyglycolic acid mesh to facilitate the tissue manipulation and implantation. An immature epithelium was obtained with a weak immunostaining to cytokeratins. The immunohistochemical study could not demonstrate the development of basement membrane. CONCLUSIONS: In vitro keratinocyte culture techniques could be applied to other epithelial tissues like the urothelium. We obtained a three-dimensional in vitro tissue suitable for grafting in a relatively short time, which needs the matrix interactions in order to mature. PMID- 16700212 TI - [Study of a intestinal enteroliths in human patient with colon adenocarcinoma. Is it similar to renal calculi?]. AB - This work shows the study performance to intestinal enterolithis from a 91 year old patient with multiple enterolithiasis confirmed by abdominal X-ray and TAC analyses showing the presence of intestinal, renal and bile stones. This enterolithis is associated with colon adenocarcinoma. The enteroliths were obtained by hemicolectomia and were analyzed by infrared spectroscopy (IR), giving non-stoichiometry carbonate apatite whitloquite-like with, possibly, organic material. By atomic emission spectroscopy we found Ca, Mg, K, Na and K (mg/100 mg of calculi) and Zn, Ba, Mn, Fe, Cu, Si, Ti and Br in minor proportion (microg/100 mg of calculi). Because of calculi morphology and the IR spectra (non stoichiometry carbonate apatite) we carried out analysis by high performance liquid chromatography (HPLC) and found coproporphyrin (about microg/g of calculi) and uroporphyrin, protoporphyrin and heptacarboxy-porphyrin in minor extent. Calculi were also studied by scanning electronic microscopy and EDX and X-ray diffraction giving crystals of CaP4O11. All these results show that intestinal enteroliths composition are similar to renal calculi although its morphology differs from renal calculi. PMID- 16700213 TI - [Laparoscopic ureterolithotomy of an iliac ureteral stone forgotten for more than eight years]. AB - INTRODUCTION: Laparoscopic ureterolithotomy recently rises as a new option in the treatment of ureteral calculi, particularly those of the greatest size, hardness or impactation. We describe such an indication to resolve a case of forgotten for more than eight years and severely obstructive ureteral stone. PATIENT AND METHOD: A 64 years-old male received extracorporeal shock wave lithotripsy for a right distal ureteral stone and, simultaneously, a left impacted iliac ureteral calculi was discovered, at the confluence of an incomplete duplication of the ureter, for which treatment was recommended, but deferred by the patient. Eight years after, the same stone caused a massive dilatation with poor function of the upper pole moiety and slightly preserved function of the lower pole moiety of the left kidney. Two intents of retrograde ureteroscopy failed because of impossibility to reach the stone. Transperitoneal laparoscopic ureterolithotomy was performed in lateral decubitus position, with double J in place and three 10 mm ports. After identification of the dilated ureter, an V-shape ureterothomy was made and the stone mobilized and extracted. The ureter was stented and the ureterothomy closed with intracorporeal suture. The patient had a postoperative stage of four days and a mild functional recovery. DISCUSSION: If the usual treatment options (extracorporeal lithotripsy and ureteroscopy with intracorporeal lithotripsy) failed, then laparoscopic ureterolithotomy is less invasive than open ureterolithotomy. However, the indications of laparoscopic ureterolithotomy are restricted because substantial laparoscopic experience is needed to cope with possible technical difficulties. PMID- 16700214 TI - [Mucinous adenocarcinoma of the urachus and peritoneal pseudomyxoma]. AB - The adenocarcinoma of the urachus is very rare tumor, with an incidence of 1/5.000.000 inhabitants, represents less than 0.001 of all types of bladder cancer. Peritoneal pseudomixoma is a rare neoplasm characterized by mucinous acites that involvement the peritoneal surface and omentum. Usually is associated with benign o malignant mucinous tumor of the appendix or ovary. In this paper, we present a case of peritoneal pseudomixoma caused by a mucinous adenocarcinoma of the urachus. PMID- 16700215 TI - [Segmental testicular infarction]. AB - We report the case of a 47 years old man previously diagnosed of left hidrocele. After having a recent mild left testicular pain, an ultrasonografic study revealed a solid hipoecoic testicular lesion rounded by a big hidrocele, suggesting a testicular neoplasm. Radical inguinal orchiectomy was made and pathologic study showed segmental testicular infarction. No malignancy was found. We review the literature of the topic. PMID- 16700216 TI - [Chronic orchialgia]. PMID- 16700217 TI - [Confirmation of the microbiological diagnosis in cases of abscess of the muscle iliopsoas]. PMID- 16700220 TI - [A extrange case of urgent urination]. PMID- 16700222 TI - Nurses debate merits of new mental health plans. PMID- 16700224 TI - A review of training is long overdue in mental health nursing. PMID- 16700225 TI - Mental health manifesto. PMID- 16700226 TI - Practice implications of the increase in measles infections. AB - The recent increase in measles and one death clearly shows it is still a serious illness and highlights the importance of high levels of vaccination uptake. This article discusses measles infection and prevention and outlines the implications for nurses. PMID- 16700227 TI - Stevens-Johnson syndrome. PMID- 16700228 TI - Homeostasis. Part 4: fluid balance. PMID- 16700229 TI - An overview of steroid use and its potential side-effects. AB - Steroids are prescribed for their physiological and pharmacological effects: they have an antiinflammatory influence and suppress immune responses. However, this can cause side-effects. Assessment for the development of side-effects and advice on avoiding infection and injury are essential aspects of the care of patients taking steroids. PMID- 16700230 TI - Improving support information with an integrated CNS service. AB - Clinical outcomes guidance highlights that a clinical nurse specialist is a necessary member of the multidisciplinary team within gynaecological oncology. Two clinical nurse specialist posts in gynaecological oncology have been developed within the North London Cancer Network. This article discusses the importance of these roles and includes results of a patient satisfaction survey. PMID- 16700231 TI - Phenytoin. PMID- 16700232 TI - Identifying the needs of carers in mental health services. AB - AIM: To assess all carers in the Gloucester Assertive Community Treatment Team and identify their needs. METHOD: Carers were identified using the definition employed in the Notional Service Framework for Mental Health (DH, 1999): they were family or close friends who spent a minimum of 12 hours per week helping/supporting the service user. Paid carers were excluded. A standardised carer's assessment that had been developed as part of the care programme approach (CPA) was used to evaluate the needs of carers. RESULTS: The study highlights the type of support that carers give those who use the mental health services and reveals the impact that this caring has on them. The carers' needs are identified as: someone to talk to; help with cleaning/ironing; help with finances; support to enable them to take a break; plus respite care and accommodation for the service user. CONCLUSION: Carers play an important role in supporting people who are diagnosed with severe mental illness although further research is necessary to consider the benefits of a closer relationship between the carer, service user and mental health professionals. PMID- 16700233 TI - Acute care in the community. Interview by Kathryn Godfrey. PMID- 16700234 TI - Lactose intolerance in infants. AB - Cathy Taylor describes the pathophysiology and aetiology of lactose intolerance and how to diagnose and treat it. Management of the infant by the primary health care team is discussed, with emphasis on advice and nutritional support that can be recommended to parents. PMID- 16700235 TI - A behavioural approach to weight loss. PMID- 16700236 TI - Management of typhoid and paratyphoid fevers. AB - Gladys Xavier discusses typhoid and paratyphoid fevers, the risks they pose to UK citizens and the role of primary healthcare professionals in their prevention and management. PMID- 16700237 TI - Why immunisation is essential for controlling yellow fever. AB - Joyce Skeet outlines the causes of yellow fever and the course of the disease, emphasising the importance of immunisation to control it. She also discusses issues that need to be considered when yellow fever vaccination is provided in primary care. PMID- 16700238 TI - How to keep up to date with practice. PMID- 16700239 TI - Gaining consent for postmortems. Interview by Clare Lomas. PMID- 16700241 TI - Fostering a nurse, patient, and family 'partnership in care'. PMID- 16700242 TI - The patient's right to choice. PMID- 16700243 TI - Use of insulin pumps for diabetes management. AB - Patients are diagnosed with diabetes mellitus in the United States at increasing rates. Nurses must maintain current knowledge of effective diabetes management strategies. An overview of the use of insulin pumps is provided, including the function, criteria for use, advantages, disadvantages, and nursing management considerations. PMID- 16700244 TI - Assessment and discharge planning for hospitalized older adults with delirium. AB - A description of the effects of delirium on attention is provided. Nursing implications pertinent to appropriate assessment for attention deficits, interventions to maintain patient safety, and discharge planning for the older adult with persistent delirium are also discussed. PMID- 16700246 TI - Nasogastric tubes: an historical context. AB - Nurses commonly care for patients with nasogastric tubes. Many authors have addressed the management of nasogastric tubes; however, discussion rarely considers the historical perspective. The purpose of this article is to provide an historical context on nasogastric tubes. PMID- 16700245 TI - Theories of aging as basis for assessment. AB - Based on biopsychosocial theories of aging, a tool was developed to assist nurses in conducting holistic adult admission assessments. The Adult Assessment Tool can facilitate comprehensive, best-practice decisions in caring for hospitalized middle-aged and older adults. PMID- 16700247 TI - A pilot study evaluating two alternate methods of stool collection for the fecal occult blood test. AB - The annual fecal occult blood test is an inexpensive colorectal cancer screening regimen effective in reducing morbidity and mortality. Researchers evaluated patient preferences and results for toilet tissue smears and direct smears, compared with the traditional wooden stick smear method. PMID- 16700248 TI - The basics of human subjects protection. PMID- 16700249 TI - Nursing research journal clubs: bridging the gap between practice and research. PMID- 16700250 TI - Presenteeism: sick and tired at work. PMID- 16700252 TI - Preventing aspiration in older adults with dysphagia. PMID- 16700251 TI - So, what do I put on this wound? Making sense of the wound dressing puzzle: Part I. PMID- 16700253 TI - Megatrends, the annual report, possibilities. AB - The successful leader and manager have two very important challenges: to scan the horizon and not be thrown into crisis by not seeing predictable surprises, and to recognize the longer, more pervasive trends that are in the more distant future. In reality, these two processes merge and drive the future planning of the patient care enterprise. The merger of the operational imperatives and the long range trends that create very different ways for us to work is the work of leaders. In the words of Senge and colleagues (2004), we are at the end of the era of hierarchical leadership and at the beginning of distributed, shared networks where "...becoming a real human being really is the primary leadership issue of our time" (p. 186). As we finish the year and begin a new one, it is time to look backward, look forward, and "see" the new world. We must also look into ourselves to see the inner work we need to do to meet the challenges of the next 5 years. PMID- 16700254 TI - The Lourdes Hospital inquiry: its implications for medical practice. PMID- 16700255 TI - Inguinal hernias in men with few symptoms: does everyone need a repair? PMID- 16700256 TI - The Snodgrass repair: is stenting always necessary? AB - To evaluate the need for stenting in Snodgrass hypospadias repairs. Sixty-five boys underwent hypospadias repairs between April 1996 and July 2001. A variety of techniques were employed. Snodgrass repair was performed in 39 patients, MAGPI in 18, Glanular approximation procedures in 4, Mathieu procedures in 3 and one Duckett Onlay-flap. Seventeen patients, all Snodgrass repairs, had placement of a urethral stent for one week postoperatively. Follow-up at six weeks and one year or when toilet-trained was carried out on all patients. A Fisher's exact test was performed analysing the difference in outcome of the distal Snodgrass repairs depending on the presence or absence of a urethral stent. The patient age at the time of surgery ranged from 8-115 months, with a median of 21 months. Postoperative complications included 1 stent migration, 1 urethral diverticulum and 4 fistulas. Also in one patient part of the ventral skin flap on one side sloughed away, this was debrided and allowed to close by secondary intention. The fistula rate in the entire group is 6%, with an incidence of 10.5% in patients undergoing Snodgrass repair. In the cases having Snodgrass repair for meatal position other than mid- or proximal shaft there was no statistical difference in the incidence of fistula whether or not a stent was used. The long-term follow-up of the entire group indicates that 5 patients developed meatal narrowing, 3 in stented Snodgrass repairs, one patient who had a MAGPI and one GAP patient. Otherwise the long-term outcome of the surgery has been excellent for the entire group. We believe that stenting can be abandoned in distal repairs without compromising patient outcomes. PMID- 16700257 TI - General practice out-of-hours co-operatives--population contact rates. AB - Since 1998, Irish general practice has developed 11 out-of-hours co-operatives, covering almost 40% of the population. The co-operatives vary in terms of triage mechanisms, treatment centres and domiciliary visits. Out-of-hours consultation rates for the GMS sector of the population (one-third of the population who receive free primary care on the basis of low income) have increased rapidly to 438 consultations/1000 persons/year by 2003. British and Danish out-of-hours co operatives report annual contact rates of 280-470 contacts/1000 persons per year. The aims were to describe 12 month activity data in the co-operatives and to describe the workload in the context of the population served. A questionnaire survey for a 12 month period was completed by all 11 co-operatives. The results were that almost 340,000 contacts occurred during the period, with 34.0% dealt with by phone advice alone, 53.8% dealt with by visits to treatment centres and 12.3% dealt with by domiciliary visits. The mean population contact rate is 221 contacts/1000 persons/year (range 370-70) and the mean consultation rate is 144 consultations/1000 persons/year. Two distinct bands of contact rates emerged - seven of eight rural co-operatives (all with domiciliary services) have a range of 220-300 contacts/1000 persons/year while three urban co-operatives (none of which have integrated domiciliary services) have a range of 70-90 contacts/1000 persons/year. These results are explored in the context of UK and Danish data, with which they compare. The implications of the urban/rural banding are significant and require early further research. PMID- 16700258 TI - Audit of Mobile Intensive Care Ambulance Service. AB - The transfer of critically ill patients between hospitals carries significant risk. The Mobile Intensive Care Ambulance Service (MICAS) in Ireland was established in 1996 to provide a ground transfer service between hospitals for critically ill patients. The aim of this audit was to determine if a centralised retrieval system for interhospital transport of critically ill patients in Ireland by MICAS is safe and efficient. 484 critically ill patients transported by MICAS between April 1996 and December 2004 were studied retrospectively. There were no patient deaths during transfer. 47% of transfers were between hospitals in the greater Dublin area. 45% were from regional centres to Dublin. 93% of patients required airway intervention before transfer (intubation or tracheostomy) and 32% needed inotropic support during transfer. The MICAS data confirm the demand for interhospital transport of critically ill patients and suggest that transfer using a centralised retrieval system is safe. PMID- 16700259 TI - Poor uptake of reproductive health screening services by female renal transplant recipients. AB - Women with functioning renal transplants are a high-risk group for de novo malignancies and other gynaecological health problems. The objective of this study was to assess patients' awareness of gynaecological issues, and to assess uptake of cervical and breast cancer screening services. A structured questionnaire on family planning, menopausal issues and knowledge/use of cervical and breast cancer screening was administered to 64 female renal transplant recipients. 58 (91%) responded to the questionnaire. Mean age at first transplantation was 35 years (range 11 - 69). 84% were aware as to why they should have regular cervical smears. 15 (26%) had, however, never had a smear and only 9 (16%) were having yearly smears. 12 of 28 postmenopausal women entered the menopause under the age of 41 years, but only 5 of these had received Hormone Replacement Therapy. Breast self examination is practiced by 71%, but only 26% have had mammograms. These figures suggest that female renal transplant patients are not adequately screened for cervical and breast cancer. The results also indicate a need for further education regarding family planning issues and menopausal health concerns. We conclude that formal gynaecological review should be routinely available for women with renal transplants. PMID- 16700260 TI - Blood alcohol levels in persons who died from accidents and suicide. AB - Although it is known that alcohol is associated with a high proportion of fatal accidents and suicides, little information is available in Ireland on blood alcohol concentrations (BACs) of those who died. This study was undertaken to identity the (BACs) in persons who died as a result of suicide or injury. The study was a retrospective review of coroners' records to identify BACs in three counties in Ireland. All cases where the person died as a result of injury or suicide in 2001 and 2002 were included. There were 129 deaths eligible for inclusion. Of these, 98 (76%) were male, 55 (42.6%) were road traffic accidents (RTAs), 31 (24.0%) suicides, 12 (9.3%) substance misuse, 11 (8.5%) house fires and 20 (15.5%) others. Of the 55 who died as a result of RTAs, 22 (40%) had positive BACs ranging from 16mg/100 ml to 325 mg/100 ml. Of the 31 who died as a result of suicide, 28 (90.3%) were male. BACs were available for 29 (93.5%). Of these, 16 (55.5%) had alcohol detected. Persons aged less than 30 years were more likely to have alcohol in their blood (p < 0.002). The mean BAC for persons aged less than 30 was 191.5 mg/100 ml compared to 84.0 mg/100 ml for those aged 30 and over. The mean BAC for adults who died in house fires was 225.2 mg/100 ml. The high BACs in those who died as a result of suicide or injury reflect the high level of alcohol consumption and binge drinking in Ireland. PMID- 16700261 TI - Cystic fibrosis-related diabetes in adults. AB - We aimed to examine the differences between patients with cystic fibrosis-related diabetes (CFRD), and those with normal glucose handling in adults with cystic fibrosis (CF) in Ireland. We conducted a retrospective analysis of patients who attend the national referral centre for adult CF. Patients were diagnosed as having CFRD by the American Cystic Fibrosis Foundation criteria for diagnosis of CFRD. Of 259 patients, 150 were classifiable and 81 (54%) were classified as having CFRD. The groups with and without CFRD were not significantly different with regard to age (median 28.4 vs 26.0 years), sex (males 56% vs 55%) or BMI (median 20.9 vs 21.3 kg/m2). The group with CFRD had poorer lung function (mean % predicted FEV1 49.9 vs 66.4, P < 0.001), poorer bone mineral density (T-scores at the lumbar spine -1.95 vs -1.44, P < 0.05 and femur -1.19 vs -0.57, P < 0.01) and a greater proportion of PSEUDOMONAS AERUGINOSA positive sputum cultures (82.5% vs 64.2%, P < 0.05). No patients with CFRD carried the R1 17H mutation whilst 19% of the group without CFRD were heterozygous for this defect (P < 0.001). In conclusion, CFRD was highly prevalent in adults. The presence of CFRD was associated with poorer lung function, poorer bone mineral density and an increased prevalence of PSEUDOMONAS AERUGINOSA in sputum. The R1 17H mutation may be protective for CFRD. PMID- 16700263 TI - Percutaneous recanalisation of two total coronary artery occlusions: an alternative option to surgical revascularisation. PMID- 16700262 TI - Molecular epidemiologic investigation of tuberculosis in an area of increasing incidence in inner-city Dublin. AB - In 2002 the tuberculosis (TB) incidence in Dublin was 13.1 cases per 100,000 persons. In inner city Dublin, the TB incidence was > 20 per 100,000 persons. TB notification rates have remained elevated. The objective of the study is to characterize the population affected in this inner city area and to investigate transmission patterns and potential risk factors for TB disease. This hospital based molecular epidemiologic study included all culture positive cases of TB between January 1, 1998 and December 31, 2002 from the Mater Misericordiae University Hospital. 142 cases were analyzed. 84 (59%) had clustered Mycobacterium tuberculosis (Mtb) isolates, 58 (41%) had unique isolates by molecular typing. There were 13 clusters; the largest two clusters had 26 cases each, one linked to pubs, the other linked through family contact. In multivariate logistic regression analysis, birth in Ireland, pulmonary TB, and younger age were significantly associated with recent transmission. Dublin is a developed city with persistent areas of high rates of TB in the native population. Despite a functioning public health system and lack of predisposing risk factors such as HIV, immigration, and extreme poverty, TB can persist in pockets of the younger, native population. PMID- 16700264 TI - Parasuicide assessment in the emergency department. PMID- 16700266 TI - 50-state roundup: ensuring children with disabilities a free appropriate public education. PMID- 16700267 TI - Legislation & regulations. Federal actions. State actions. PMID- 16700268 TI - Patient factors influencing restoration longevity. PMID- 16700269 TI - Facts and fallacies: an evidence base for complete dentures. AB - The rate of edentulism is falling but many edentulous people still need complete dentures. There is poor correlation between the quality of dentures and patient satisfaction. There is no evidence that a more complex fabrication technique including face-bow registration results in a better clinical outcome. Balanced occlusion/articulation is not necessary for successful complete denture function. Variations in materials and techniques appear to have only minor influence, if any, on the clinical end result. Psychosocial factors, especially a good relationship between dentist and patient, are more important than prosthodontic factors for a positive outcome. CLINICAL RELEVANCE: Creating a good relationship between the dentist and the patient is extremely important for a successful outcome of a complete denture treatment. PMID- 16700270 TI - Current clinical applications of photo-activated disinfection in restorative dentistry. AB - The traditional treatment of dental caries has been limited to bulk removal of the diseased tissue. Recently, disinfection rather than the removal of all the carious tissue has been advocated with the aim of more effective and conservative treatment. In endodontics, prognosis is dependent on the complete disinfection of the root canal system. Evidence exists that Photo-Activated Disinfection is more effective than traditional chemo-mechanical canal preparation. Photo-Activated Disinfection may also have a place in the treatment of other infective oral conditions, including periodontal disease. CLINICAL RELEVANCE: Removal or disruption of micro-organisms and their ecosystem is important in many branches of restorative dentistry. A novel disinfection system is now available which has the potential to improve treatment methods and prognoses. PMID- 16700271 TI - Self-inflicted dental injury presenting as localized anterior tooth surface loss. AB - A case of localized anterior tooth surface loss (TSL) with an unusual aetiology is reported. Whilst suffering from a bout of acute depression and anxiety, a 29 year-old female caused significant trauma to her anterior dentition with a pair of fabric scissors. The presentation and management of this case is described. CLINICAL RELEVANCE: Although tooth surface loss is a common clinical finding in many patient groups, practitioners should be aware of possible unusual aetiologies which may be involved. PMID- 16700272 TI - Manual versus powered toothbrushes: what is the evidence? AB - Powered toothbrushes are a significant segment of the oral care market. Numerous clinical trials have compared the effectiveness of manual and powered toothbrushes for their effectiveness in improving oral health and the results are often conflicting. Recent Systematic Reviews by the Cochrane Oral Health Group have distilled this information and provided unbiased conclusions, namely that: powered toothbrushes with an oscillating rotating action are more effective than manual toothbrushes in reducing plaque and gingivitis; other types of powered toothbrushes produced less consistent reductions in plaque and gingivitis than manual brushes. Further studies are required to determine the relative effectiveness of manual and powered toothbrushes in improving the status of patients with periodontitis. CLINICAL RELEVANCE: This review provides the dental professional with an unbiased summary of current evidence regarding the relative effectiveness of manual and powered toothbrushes. PMID- 16700273 TI - A review of the current uses of Botox for dentally-related procedures. AB - Botox is the most widely used abbreviation for botulinum toxin type A (BtA). This drug has received wide coverage in the press and media for its cosmetic advantages, especially in the facial region. The clinician may be aware of the many courses becoming available and aimed at dentists to start using it in the cosmetic context. This paper intends to provide a basic understanding of the many functional uses of the drug in the orofacial region that may be relevant to everyday practice. CLINICAL RELEVANCE: Awareness of the current medical, as opposed to purely cosmetic, uses of botulinum toxin in the orofacial region is desirable and this paper should inform clinicians and their patients with respect to this increasingly popular drug. PMID- 16700274 TI - Surgical emphysema following a restorative procedure: A case report. AB - Surgical emphysema is a rare complication following a restorative procedure, despite the use of air-driven handpieces. Although most cases resolve spontaneously, it should be correctly managed to avoid potentially life threatening situations, such as pneumomediastinum and pneumothorax. CLINICAL RELEVANCE: Surgical emphysema, although rare, can occur following certain dental procedures. It should be avoidable but, when it occurs, appropriate management is essential. PMID- 16700275 TI - Smoking cessation and the dental team. AB - Dental teams are in an ideal position to help smokers stop smoking. In spite of this, smoking cessation interventions are not often incorporated as a routine part of dental care. Surveys have shown that most dentists believe that helping patients stop smoking is important, but they are reluctant to provide smoking cessation services for their patients. However, comparatively little research has been carried out to examine the views and activities of other members of the dental team, with respect to the provision of smoking cessation support as part of dental care.This paper describes a self-administered, questionnaire-based study to investigate attitudes and practices amongst staff in the Birmingham Personal Dental Service, with respect to helping patients to stop smoking. CLINICAL RELEVANCE: Dentists and their team members have an important role to play in smoking cessation. PMID- 16700276 TI - Renal cell carcinoma: A rare metastasis to the tongue--a case report. AB - This paper describes the diagnosis and treatment of a patient who presented with a lesion on the left posterior scalp, who was also experiencing more generalized signs of malaise. Biopsy led to the diagnosis of renal cell carcinoma and additional lesions developed in the tongue. About 1% of all malignancies found in the oral cavity are metastatic. CLINICAL RELEVANCE: Metastases to the head and neck are uncommon but should always be considered among the differential diagnoses of lesions. PMID- 16700277 TI - Physical signs for the general dental practitioner. Immune thrombocytopenic purpura. PMID- 16700279 TI - Moral agency in pain medicine: philosophy, practice and virtue. AB - Although established as a field of specialization, pain medicine remains somewhat fractionated. Such lack of cohesion creates dissonance on multiple levels, and thus, impedes the provision of effective pain care. This paper asserts that there is a core philosophy of medicine that reflects the intellectual and moral quality of the healing relationship. I argue that pain medicine, in all its constituent disciplines, is bound to this philosophy. The intricate relationship between pain, the pain patient, and the pain physician creates pragmatic and moral dilemmas that may not be well served by the use of prima facie principles. It is argued that an agent-based, virtue ethics best enable the clinician to both apprehend the complexity of this relationship and appreciate other ethical approaches in the discourse arising from issues of care. PMID- 16700278 TI - Opioid guidelines in the management of chronic non-cancer pain. AB - BACKGROUND: Opioid abuse has increased at an alarming rate. However, available evidence suggests a wide variance in the use of opioids, as documented by different medical specialties, medical boards, advocacy groups, and the Drug Enforcement Administration (DEA). OBJECTIVES: The objective of these opioid guidelines by the American Society of Interventional Pain Physicians (ASIPP) is to provide guidance for the use of opioids for the treatment of chronic non cancer pain, to bring consistency in opioid philosophy among the many diverse groups involved, to improve the treatment of chronic non-cancer pain, and to reduce the incidence of drug diversion. DESIGN: A policy committee evaluated a systematic review of the available literature regarding opioid use in managing chronic non-cancer pain. This resulted in the formulation of the essentials of guidelines, a series of potential evidence linkages representing conclusions, followed by statements regarding relationships between clinical interventions and outcomes. METHODS: Consistent with the Agency for Healthcare Research and Quality (AHRQ) hierarchical and comprehensive standards, the elements of the guideline preparation process included literature searches, literature synthesis, systematic review, consensus evaluation, open forum presentations, formal endorsement by the Board of Directors of the American Society of Interventional Pain Physicians (ASIPP), and blinded peer review. Evidence was designated based on scientific merit as Level I (conclusive), Level II (strong), Level III (moderate), Level IV (limited), or Level V (indeterminate). RESULTS: After an extensive review and analysis of the literature, the authors utilized two systematic reviews, two narrative reviews, 32 studies included in prior systematic reviews, and 10 additional studies in the synthesis of evidence. The evidence was limited. CONCLUSION: These guidelines evaluated the evidence for the use of opioids in the management of chronic non-cancer pain and recommendations for management. These guidelines are based on the best available scientific evidence and do not constitute inflexible treatment recommendations. Because of the changing body of evidence, this document is not intended to be a "standard of care." PMID- 16700280 TI - The effect of sedation on diagnostic validity of facet joint nerve blocks: an evaluation to assess similarities in population with involvement in cervical and lumbar regions (ISRCTNo: 76376497). AB - BACKGROUND: Zygapophysial or facet joint pain in patients suffering with chronic spinal pain without disc herniation or radiculopathy may be diagnosed with certainty by the use of controlled diagnostic blocks. But, in patients suffering with either lumbar or cervical facet joint pain, even this diagnostic approach may be confounded by false-positives when using a single diagnostic block. It may also be confounded by the administration of anxiolytics and narcotics prior to, or during, the controlled diagnostic facet joint blocks. The effect of sedation on the validity and potential differential results in patients suffering with combined cervical and lumbar facet joint pain has not been evaluated. OBJECTIVE: To assess the effects of midazolam and fentanyl on the diagnostic validity of facet joint blocks in patients suffering with both cervical and lumbar facet joint pain. STUDY DESIGN: Randomized, double-blind, placebo-controlled study. METHODS: The design consisted of a placebo group receiving a sodium chloride solution and two experimental groups receiving either midazolam or fentanyl. Patients included in the study had been diagnosed with facet joint pain using controlled comparative local anesthetic blocks of the medial branches and L5 dorsal rami. They had been treated with lumbar and cervical facet joint nerve blocks and experienced good pain relief; and were presenting for repeat treatment after a period of symptom relief. The study was performed in an interventional pain management practice in the United States; a total of 60 patients participated with 20 patients randomly allocated into each group. Outcome measures included numeric pain scores, proportion of pain relief, and ability to perform prior painful movements. OUTCOME MEASURES: Outcomes were assessed at baseline and after the administration of 1 of the 3 solutions (Group I, sodium chloride solution; Group II, midazolam; or Group III, fentanyl). RESULTS: Overall, 50% of the patients were relaxed or sedated in the placebo group, while 100% of the patients in the midazolam and fentanyl groups were relaxed or sedated. As many as 10% of the patients reported significant relief (>= 80%) with the ability to perform prior painful movements. CONCLUSIONS: Perioperative administration of sodium chloride, midazolam, or fentanyl can confound results in the diagnosis of combined cervical and lumbar facet joint pain. False-positive results with placebo or sedation may be seen in a small proportion of patients. PMID- 16700281 TI - Lumbosacral transitional vertebrae: incidence in a consecutive patient series. AB - BACKGROUND: Great variability exists in the reported frequency of transitional vertebra in the general population as well as in symptomatic lumbar spine patients. OBJECTIVE: To determine the occurrence rate of transitional vertebra in consecutive lumbar spine patients in our practice. DESIGN: A prospective, observational study METHODS: Plain films were viewed and the presence or absence of a lumbar transitional vertebra was documented. Transitional vertebrae were classified as incomplete or complete. RESULTS: Of the 300 consecutive patients, 30% had a transitional vertebra. CONCLUSION: Our percentage of patients with a transitional vertebra was higher than the percentages reported for the general population. This supports our belief that the alteration in mechanics caused by a transitional vertebra may at times contribute to pain generation. Additionally, as a transitional segment may not always be visualized on MRI, obtaining plain films prior to an injection or surgery will help prevent confusion over the appropriate level for the procedure. PMID- 16700282 TI - Does adherence monitoring reduce controlled substance abuse in chronic pain patients? AB - BACKGROUND: Opioids are used extensively for chronic pain management in the United States. The frequency of opioid use prior to presenting to interventional pain management settings and in interventional pain management settings has been shown to be above 90%. Given that controlled substance abuse and illicit drug use are prevalent phenomena, adherence monitoring of patients that are prescribed opioids is becoming common. Adherence monitoring is carried out by an appropriate history, periodic evaluation of appropriate intake of drugs, random drug testing, and pill counts. Crucial to adherence monitoring is an initial controlled substance agreement and repeated review of the terms of this agreement with on going education. However, the effect of adherence monitoring on drug abuse is unclear. OBJECTIVE: To identify controlled substance abuse through implementation of the terms of a controlled substance agreement, including periodic review and monitoring outside the organization. STUDY DESIGN: Prospective evaluation with historical controls. METHODS: Five hundred consecutive patients receiving prescription controlled substances were followed in a prospective manner. The evaluation consisted of a chart review to monitor controlled substance intake, with special attention to drugs obtained from outside the organization. Data collection for this purpose included information from records, pharmacies, referring physicians, and all the physicians involved in the treatment of the patient. RESULTS: Results from 500 consecutive patients were evaluated. Controlled substance abuse was seen in 9% of patients; overall, 5% of patients were obtaining controlled substances from other physicians, and 4% from illegal sources. CONCLUSION: Adherence monitoring, including controlled substance agreements and various periodic measures of compliance was associated with a 50% reduction in opioid abuse. PMID- 16700283 TI - The sacroiliac joint: anatomy, physiology and clinical significance. AB - The sacroiliac joint (SIJ) is a putative source of low back pain. The objective of this article is to provide clinicians with a concise review of SIJ structure and function, diagnostic indicators of SIJ-mediated pain, and therapeutic considerations. The SIJ is a true diarthrodial joint with unique characteristics not typically found in other diarthrodial joints. The joint differs with others in that it has fibrocartilage in addition to hyaline cartilage, there is discontinuity of the posterior capsule, and articular surfaces have many ridges and depressions. The sacroiliac joint is well innervated. Histological analysis of the sacroiliac joint has verified the presence of nerve fibers within the joint capsule and adjoining ligaments. It has been variously described that the sacroiliac joint receives its innervation from the ventral rami of L4 and L5, the superior gluteal nerve, and the dorsal rami of L5, S1, and S2, or that it is almost exclusively derived from the sacral dorsal rami. Even though the sacroiliac joint is a known putative source of low back and lower extremity pain, there are few findings that are pathognomonic of sacroiliac joint pain. The controlled diagnostic blocks utilizing the International Association for the Study of Pain (IASP) criteria demonstrated the prevalence of pain of sacroiliac joint origin in 19% to 30% of the patients suspected to have sacroiliac joint pain. Conservative management includes manual medicine techniques, pelvic stabilization exercises to allow dynamic postural control, and muscle balancing of the trunk and lower extremities. Interventional treatments include sacroiliac joint, intra-articular joint injections, radiofrequency neurotomy, prolotherapy, cryotherapy, and surgical treatment. The evidence for intra-articular injections and radiofrequency neurotomy has been shown to be limited in managing sacroiliac joint pain. PMID- 16700284 TI - An electrophysiological approach to the evaluation of regional sympathetic dysfunction: a proposed classification. AB - BACKGROUND: The importance to physicians of maintaining a level of understanding of illnesses and their treatment continues to reveal itself in a most striking fashion when it comes to the progressive interest recently directed to disorders of the autonomic nervous system (ANS). In particular, the relevance to pain practitioners of disease states which directly involve the sympathetic portion of the ANS has increased markedly following the international renaming of reflex sympathetic dystrophy (RSD) and causalgia to complex regional pain syndrome (CRPS) Type I and Type II respectively, as well as sympathetically maintained pain (SMP). Subsequently it has become better understood that many other forms of neuropathic pain also demonstrate local abnormalities of the sympathetic nervous supply to the skin within the painful territory, thereby increasing the diagnostic value of these (often subtle) cutaneous clinical signs. OBJECTIVES: The objectives of this presentation include (a) a concise review of laboratory tests that are currently used in the evaluation of the autonomic nervous system, (b) a discussion of those procedures that were developed for the assessment of sympathetic sudomotor function, (c) a review of the anatomic pathways subserving those electrophysiological methods for sudomotor testing, and (d) the current diagnostic classification for regional abnormalities of sympathetic sudomotor dysfunction. METHODS: Methods used in the preparation of this article have included a review of (a) historic clinical and laboratory articles (or translations thereof) regarding the medical importance of disorders of the autonomic nervous system, dating back to more than 155 years ago (b) anatomic and electrophysiological basis for electroneurodiagnostic sudomotor testing, and (c) the author's proposal for a diagnostic classification of regional sympathetic sudomotor dysfunction. PMID- 16700285 TI - Bayes window (4): table of illusions. PMID- 16700286 TI - Lightness and perceptual transparency. AB - To estimate intrinsic descriptors of objects in the environment, effective biological vision systems must 'discount' extrinsic image properties that arise from changes in viewing conditions. In particular, to estimate the reflectance of surfaces, human vision must discount, or 'take account of', likely differences in the illumination of surfaces between one image region and another. If human vision possesses any significant degree of lightness constancy, then we would expect a target perceived to be in low illumination to appear lighter than an identical target perceived to be in higher illumination. In this paper, I present lightness illusions that run directly counter to this expectation. I suggest that mid-level and higher-level factors such as image junction structure and perceived illumination and transparency, are ineffective for generating strong lightness illusions on their own, and that these factors are not 'stronger' than luminance contrast in determining lightness. I discuss the implications of these results for current models of lightness perception. I also suggest a statistical justification for the highest-luminance anchoring rule for lightness. PMID- 16700287 TI - Simultaneous lightness contrast on plain and articulated surrounds. AB - Simultaneous lightness contrast is stronger when the dark and light backgrounds of the classic display (where one of the targets is an increment and the other is a decrement) are replaced by articulated fields of equivalent average luminances. Although routinely attributed to articulation per se, this effect may simply result from the increase in highest luminance in the light articulated, vs plain, background; by locally darkening the decremental target, such an increase would amplify the difference between the targets. We disentangled the effects of highest luminance and articulation by measuring, separately, the magnitude of lightness contrast on dark and light plain and articulated backgrounds. We found that highest luminance and articulation contribute separately to the final illusion. PMID- 16700288 TI - Visual transients reveal the veridical position of a moving object. AB - The position of a moving object is often mislocalised in the direction of movement. At the input stage of visual processing, the position of a moving object should still be represented veridically, whereas it should become closer to the mislocalised position at a later processing stage responsible for positional judgment. Here, we show that visual transients expose the veridical position of a moving object represented in early visual areas. For example, when a ring is flashed on a moving bar, the part of the bar within the ring is perceived at the veridical position, whereas the part outside the ring is perceived to be ahead of the ring as in the flash-lag effect. Our observations suggest that a filling-in process is triggered at the edges of the flash. This indicates that, in early cortical areas, moving objects are still represented at their veridical positions, and the perceived location is determined by the higher visual areas. PMID- 16700290 TI - 'Squaring' is better at predicting plaid motion than the vector average or intersection of constraints. AB - How do humans combine the velocity information from two moving gratings (plaids) to detect pattern motion direction?-We are still unable to answer this question. The 'intersection of constraints' rule (IOC-Adelson and Movshon, 1982 Nature 300 523-525), and the 'vector average' rule (VA-Wilson et al, 1992 Visual Neuroscience 9 79-97) have both been supported by results in the plaid literature, but could these results be predicted by a 'squaring' nonlinearity that now forms part of several influential spatiotemporal energy models (Wilson et al 1992, loco cit.; Lu and Sperling, 1995 Vision Research 35 2697-2722; Simoncelli and Heeger, 1998 Vision Research 38 743-761)? Spatiotemporal energy in these models predicts directions other than those predicted by standard spatiotemporal energy models and may underlie the results that support the combination rules. The two combination rules and predictions from 'squaring' were tested under identical conditions. In the first three experiments a plaid was randomly presented in one of 45 different orientations, and observers were asked to remember the direction. The stimulus was then replaced by an oriented line indicating the direction predicted by one of the hypotheses. The observer was unaware which hypothesis had generated the line and was asked to make a same/different judgment. Results showed that the 'squaring' hypothesis was better at predicting perceived direction than either the IOC or VA. PMID- 16700289 TI - Spatial dependence of color assimilation by the watercolor effect. AB - Color assimilation with bichromatic contours was quantified for spatial extents ranging from von Bezold-type color assimilation to the watercolor effect. The magnitude and direction of assimilative hue change was measured as a function of the width of a rectangular stimulus. Assimilation was quantified by hue cancellation. Large hue shifts were required to null the color of stimuli < or = 9.3 min of arc in width, with an exponential decrease for stimuli increasing up to 7.4 deg. When stimuli were viewed through an achromatizing lens, the magnitude of the assimilation effect was reduced for narrow stimuli, but not for wide ones. These results demonstrate that chromatic aberration may account, in part, for color assimilation over small, but not large, surface areas. PMID- 16700291 TI - Grouping and trajectory storage in multiple object tracking: impairments due to common item motions. AB - In our natural viewing, we notice that objects change their locations across space and time. However, there has been relatively little consideration of the role of motion information in the construction and maintenance of object representations. We investigated this question in the context of the multiple object tracking (MOT) paradigm, wherein observers must keep track of target objects as they move randomly amid featurally identical distractors. In three experiments, we observed impairments in tracking ability when the motions of the target and distractor items shared particular properties. Specifically, we observed impairments when the target and distractor items were in a chasing relationship or moved in a uniform direction. Surprisingly, tracking ability was impaired by these manipulations even when observers failed to notice them. Our results suggest that differentiable trajectory information is an important factor in successful performance of MOT tasks. More generally, these results suggest that various types of common motion can serve as cues to form more global object representations even in the absence of other grouping cues. PMID- 16700292 TI - Structural similarity and spatiotemporal noise effects on learning dynamic novel objects. AB - The spatiotemporal pattern projected by a moving object is specific to that object, as it depends on both the shape and the dynamics of the object. Previous research has shown that observers learn to make use of this spatiotemporal signature to recognize dynamic faces and objects. In two experiments, we assessed the extent to which the structural similarity of the objects and the presence of spatiotemporal noise affect how these signatures are learned and subsequently used in recognition. Observers first learned to identify novel, structurally distinctive or structurally similar objects that rotated with a particular motion. At test, each learned object moved with its studied motion or with a non studied motion. In the non-studied motion condition we manipulated either dynamic information alone (experiment 1) or both static and dynamic information (experiment 2). Across both experiments we found that changing the learned motion of an object impaired recognition performance when 3-D shape was similar or when the visual input was noisy during learning. These results are consistent with the hypothesis that observers use learned spatiotemporal signatures and that such information becomes progressively more important as shape information becomes less reliable. PMID- 16700293 TI - An investigation of perceptual and decisional influences on the perception of hierarchical forms. AB - When observers are asked to identify the global and local dimensions of hierarchical forms, their responses are typically faster when the dimensions are consistent rather than inconsistent. This effect, which we refer to as the dimensional consistency effect, has been demonstrated numerous times in paradigms requiring responses to a single dimension. However, most hypotheses regarding dimensional consistency effects address the simultaneous perception of both dimensions, and the manner in which the information about these dimensions may (or may not) 'interact'. Most explanations of the dimensional consistency effect attribute the effect to perceptual influences. The present study uses the constructs of general recognition theory (Ashby and Townsend, 1986 Psychological Review 93 154-179) to represent the source of the effect in terms of both perceptual and decisional influences. In addition, a complete identification response paradigm was used to collect data on the simultaneous perception of both global and local dimensions of hierarchical forms. Analyses of multidimensional signal detection measures and fits of parametric models of the perceptual space were used to guide inferences, with all sources of evidence pointing to both perceptual and decisional influences. PMID- 16700294 TI - Configural information in gender categorisation. AB - The role of configural information in gender categorisation was studied by aligning the top half of one face with the bottom half of another. The two faces had the same or different genders. Experiment 1 shows that participants were slower and made more errors in categorising the gender in either half of these composite faces when the two faces had a different gender, relative to control conditions where the two faces were nonaligned or had the same gender. This result parallels the composite effect for face recognition (Young et al, 1987 Perception 16 747-759) and facial-expression recognition (Calder et al, 2000 Journal of Experimental Psychology: Human Perception and Performance 26 527-551). Similarly to responses to face identity and expression, the composite effect on gender discrimination was disrupted by inverting the faces (experiment 2). Both experiments also show that the composite paradigm is sensitive to general contextual interference in gender categorisation. PMID- 16700295 TI - Font tuning associated with expertise in letter perception. AB - Font tuning (FT) occurs when observers recognize a sequence of letters presented in the same font faster than in different fonts (Sanocki 1987, 1988 Journal of Experimental Psychology. Human Perception and Performance 13 267-278; 14 472 480). Here, we test the hypothesis that FT is associated with expertise with a specific writing system. We developed a systematic search task allowing the measurement of FT over a large number of letters and generalized the finding of FT from English readers viewing Roman letters to Chinese readers viewing Chinese characters. Non-Chinese readers did not show evidence of FT for Chinese characters in this search task. We also used a simpler 3-letter identification task to directly compare novice and expert readers, and to explore FT for different aspects of font such as fill, slant, and aspect ratio. We found that experts tune to aspect ratio but not to the other font changes. These findings reveal that letters are not processed visually in the same manner as shapes, and suggest an explanation for the cortical specialization obtained in the visual system for letters. PMID- 16700296 TI - Large sex difference in adolescents on a timed line judgment task: attentional contributors and task relationship to mathematics. AB - Visuospatial performance, assessed with the new, group-administered Judgment of Line Angle and Position test (JLAP-13), varied with sex and mathematical competence in a group of adolescents. The JLAP-13, a low-level perceptual task, was modeled after a neuropsychological task dependent upon functioning of the posterior region of the right hemisphere [Benton et al, 1994 Contributions to Neuropsychological Assessment: A Clinical Manual (New York: Oxford University Press)]. High-school boys (N = 52) performed better than girls (N = 62), with a large effect for sex (d = 1.11). Performance increased with mathematical competence, but the sex difference did not vary significantly across different levels of mathematics coursework. On the basis of earlier work, it was predicted that male, but not female, performance in line judgment would decline with disruptions to task geometry (page frame), and that the sex difference would disappear with disruptions to geometry. These predictions were supported by a number of univariate and sex-specific analyses, although an omnibus repeated measures analysis did not detect the predicted interaction, most likely owing to limitations in power. Thus, there is partial support for the notion that attentional predispositions or strategies may contribute to visuospatial sex differences, with males more likely than females to attend to, and rely upon, internal or external representations of task geometry. Additional support for this hypothesis may require development of new measures or experimental manipulations with more powerful geometrical disruptions. PMID- 16700297 TI - Active and passive movements give rise to different judgments of coldness. AB - When the right index fingertip of twelve subjects was moved across a cold (15 degrees C) tile by a machine (passive-guided condition), the subjects rated the temperature of the tile as being colder than when they moved the finger across the stimulus themselves (active condition). Results confirmed that active movements were associated with an attenuation of 'coldness'. When these findings are considered alongside those of earlier experiments (see VanDoorn et al, 2005 Perception 34 231 236), it may be concluded that intentionality of movement plays some role in this attenuation. PMID- 16700298 TI - [The evolutionist fallacy of early visitors. Analogies between 'primitive peoples' and prehistoric man in medical historiography]. AB - Accounts of 'prehistoric medicine' and 'ethnomedicine' have sometimes led to conclusions by analogy in medical historiography that are seen as highly problematic in modern cultural anthropology. However, this review of medical historical writings of the last three centuries shows that evolutionist identifications of early with foreign medicine were not a permanent trait of medical historiography. This approach flourished mainly in the climate of certain movements or periods that were characterised by fanatical belief in progress and by social utopias: the French Revolution, Darwinism and the period of industrial expansion in Germany, and National Socialism. Medical historiography shared this problematic approach with contemporary (social and cultural) anthropology, and - despite this methodological misuse - both acknowledged the legitimacy or even requirement of studying also similarities in the development of different periods and cultures. PMID- 16700299 TI - ["Treating mankind honestly": medical innovation between market, public and science around 1800]. AB - This contribution is based on a case study of Professor Gottfried Christian Reich (1769-1848) of Erlangen, who in 1799 published what he considered to be an infallible method of curing any illness associated with a fever. This paper shows how in the course of a published debate the focus shifted from a medico scientific issue to a moral one. Furthermore, the experiments carried out at Berlin's Charite hospital by order of the Prussian Medical Authority illustrate how the public debate had an effect on the assessment of the empirical evidence produced. Overall, this case study demonstrates that we can use the history of a medical innovation as a means to highlight the interconnections between medical science, the public, and the market. PMID- 16700300 TI - [Paul Ehrlich and commercial serum production: on the control of diphtheria antitoxin in the laboratory and in industry]. AB - The state control of the anti-diphtheria serum is a prime example for the efficient cooperation between state authorities, industrials and scientists. The article describes firstly the dynamics of this development and places it in the context of already existing approaches to drugs control. Of special interest are the motives which led the different actors to support the state control of sera. Secondly the reciprocal influences between Ehrlich and the serum producers are analyzed. It is shown that practical problems inspired Ehrlich's theoretical work, which in return led to an improvement of measurement techniques. PMID- 16700301 TI - ['Iatros philosophos isotheos'--Nelly Tsouyopoulos (1930-2005): philosopher of science and historian of medicine. With a bibliography of her works by Sabine Rogge and Claudia Wiesemann]. PMID- 16700302 TI - [Medicine in concentration camps: deliberate neglect, minimal care, medical crimes. A permanent exhibition at the memorial site of Sachsenhausen]. AB - In November 2004, the exhibition "Medicine and Crime" was opened at the Memorial Site of Sachsenhausen near Berlin. The exhibition is located in barracks R I and R II, which have persisted in their original form. These barracks were part of the camp infirmary, which was extended repeatedly up to the end of the war. Its medical facilities included laboratories, operating theatres and wards. The infirmary was the place where "racial research" and numerous medical experiments were carried out on inmates. SS doctors performed compulsory sterilisations and castrations. Several thousand inmates were murdered in systematically planned programmes to dispose of the sick, such as "Operation 14f 13". As shown in the exhibition, the infirmary was also meant for providing minimal medical care for inmates to prevent the spread of contagious diseases and epidemics. In 1942, after setbacks in the war, attention was paid to restoring sick inmates' ability to work. Overcrowding, inhuman treatment and poor supply of medicines led to largely disastrous conditions in the infirmary. It is a special aspect of this exhibition that the events in the infirmary are told from the perspective of the inmates, i. e. the victims, not the perpetrators. PMID- 16700303 TI - Epidemiological aspects of craniopharyngioma. AB - Craniopharyngioma is a benign intracranial tumor that can be clinically aggressive. The descriptive epidemiology of this tumor is not fully known, in part because of its borderline nature between a benign and malignant tumor, and in part because of its rarity. It has a bimodal age distribution with a peak between 5 and 14 years of age, and a second one in adults older than 65 years. The Childhood Cancer Registry of Piedmont, Italy, estimates an incidence of 1.4 cases per million children per year. Similar data are provided by other registries in Western countries, while higher rates have been observed in Asia and Africa. There are no known specific environmental risk factors for craniopharyngioma, and genetic predisposition is not demonstrated. Survival has dramatically improved in the last years. PMID- 16700304 TI - Paediatric craniopharyngioma: a pathological review. PMID- 16700305 TI - Neuroimaging of pediatric craniopharyngiomas: a pictorial essay. AB - Craniopharyngiomas are benign, partly cystic epithelial tumors that account for 5 13% of all intracranial tumors and 50% of all suprasellar masses in children. They pose several challenges to the neuroradiologist. This paper aims to focus on the main issues regarding the neuroradiological diagnosis and follow-up after treatment. Topics include (i) the imaging modalities (i.e., CT and MRI) utilized in the diagnostic workup; (ii) the neuroradiological appearance, focusing on calcifications, cystic components, contrast enhancement, and location; (iii) the relationships to adjacent structures, such as the optic chiasm, hypothalamus, third ventricle, and circle of Willis; (iv) the differential diagnosis with other tumoral and non-tumoral entities occurring in the sellar/suprasellar area; and (v) post-treatment imaging issues. PMID- 16700306 TI - Craniopharyngioma: ophthalmological aspects at diagnosis. AB - AIM: We studied ophthalmological signs at diagnosis in children with craniopharyngioma. PATIENTS AND METHODS: Twenty-nine children aged from 3 to 16 years (mean 9.1 years) were retrospectively studied. All of them had visual acuity measurement, oculomotor examination, fundoscopy and, whenever possible, visual field testing. RESULTS: Visual signs were present at diagnosis in 96% of patients in our series, but were the circumstances of diagnosis of craniopharyngioma in only 34.5%. Loss of visual acuity was present at diagnosis in 15 patients (51.7%), strabismus in eight (27.6%), papilloedema in ten (34.4%), and optic nerve atrophy in 11 patients (37.9%). A visual field defect was present at diagnosis in all 18 children who were able to perform an instrumental visual field and in three of the children who were tested by attraction visual field. CONCLUSIONS: In children, ophthalmological signs are not the main circumstance of diagnosis of craniopharyngioma because children do not complain of a mild or unilateral reduction in visual acuity. The central visual field is more often affected than the peripheral visual field; computerized static automated perimetry, which allows precise study of the central visual field, is difficult to perform in children. Because of the infiltrating nature of craniopharyngiomas, visual field defects are sometimes atypical and do not indicate a chiasmal problem. PMID- 16700307 TI - The pre-operative endocrine assessment of craniopharyngiomas. AB - Pre-operative endocrinopathies are common in patients presenting with craniopharyngiomas. Clinical features may not be obvious and careful pre operative endocrine assessment is essential. Failure to recognise and address pre operative diabetes insipidus and secondary hypoadrenalism is potentially fatal. We review the available published data on pre-operative endocrine dysfunction and suggest an approach to assessment and management before surgery. PMID- 16700308 TI - Radical resection for craniopharyngiomas in children: surgical technique and clinical results. AB - Craniopharyngiomas are benign suprasellar tumors; however, their tendency to recur after resection and the risks associated with aggressive tumor resection pose a surgical dilemma. We reviewed our series of operated craniopharyngiomas and selected pediatric cases operated as first-hand cases and followed in our institution. We studied 37 cases. Resection was total in 65%. No patient died in the operative period; three patients died 6 months to 8 years after surgery of endocrine-related cause or sudden death. Tumor progression occurred in 93% of cases after subtotal resection versus 43% after total resection. Among 20 operations for recurrence, total resection was achieved in 45%. Visual deficits were often stabilized or improved after surgery, but worsened in 30% because of surgical damage or tumor recurrence. Fifty-nine percent of patients follow a normal school curriculum. All patients have some degree of pituitary hormonal replacement and 48% have obesity. Hypothalamic damage was generally associated with intraventricular extension of the tumor recurrence and re-operations, especially through combined approaches. Craniopharyngiomas in children are particularly aggressive tumors. Although the best oncological results are obtained with total resection, in some cases the functional price of surgery may be too high and alternative techniques should be proposed. The life-long management of these patients requires that the best use is made of all potential therapeutic tools. PMID- 16700309 TI - Conservative surgical approach in treatment strategy of craniopharyngioma: experience at a single institution in Italy. PMID- 16700310 TI - The role of transventricular neuroendoscopy in the management of craniopharyngiomas: three patient reports and review of the literature. AB - BACKGROUND: Management of craniopharyngiomas is problematic and often requires multimodal protocols. In the last decade neuroendoscopy has been increasingly used in the management of these lesions. PATIENT REPORTS: We report three cases of craniopharyngiomas in which various endsocopic procedures were performed. In the first case the endoscopic approach alone allowed the gross total removal of an intraventricular craniopharyngioma; in the second case the endoscopic fenestration of a cystic intraventricular craniopharyngioma allowed long-term control of the tumor with no further treatment; in the third case endoscopic fenestration and drainage of the cystic part allowed collapse of the cyst with resolution of mass effect on the visual pathway and control of hydrocephalus. The patient later underwent microsurgical removal of the solid portion of the tumor. CONCLUSIONS: Neuroendoscopy has shown great versatility in the management of intra/paraventricular craniopharyngiomas. It can be used alone to achieve gross total removal or marsupialization of cystic tumors, or, more often, in association with additional therapies, such as microsurgery, radiosurgery or intracavitary drug administration. PMID- 16700311 TI - Surgical management of craniopharyngiomas--experience with a pediatric series. AB - Because of their peculiar location and the possible related postoperative sequelae, craniopharyngiomas usually present difficult management, especially in children. The authors report on 20 years experience with a pediatric series. Fifty-four consecutive children were operated on. The goal of surgery was total tumor removal. When pre-operative neuroimaging and/ or intra-operative findings suggested more cautious behavior, subtotal/partial excision was performed, followed by radiotherapy only in selected cases. Gross total mass removal was obtained in 78% of patients, subtotal in 17%, and partial in 5%. Recurrence rate was 7% after total removal and 50% after subtotal/ partial excision. Surgical mortality accounted for 3.7% (11% in case of re-operation). After 9 years median follow-up, up to 60% of the surviving patients rely on hormone replacement, 40% present diabetes insipidus, and 18% are obese. All but two patients enjoy a normal social life. Total surgical removal of craniopharyngiomas should be attempted in children whenever possible in order to minimize the risk of recurrence. In case of huge tumors involving the hypothalamus, a more conservative resection should be performed, followed by a 'wait and see' policy or possible radiotherapy. PMID- 16700312 TI - Multimodal treatment of craniopharyngioma: defining a risk-adapted strategy. AB - Craniopharyngiomas remain a therapeutic challenge in terms of treatment-related morbidity. The fact that they are rare, that they are histologically benign, and that anatomically they are located in a challenging area, made them worthy surgical prizes. Whilst surgery has allowed us to meet the challenge of visual salvage, and the advances in hormone replacement therapy are addressing the endocrine challenge, we are becoming increasingly aware that for many children radical resection comes with the cost of devastating effects on quality of life. Increasingly we are recognizing that the next challenge is preservation of hypothalamic function. This consciousness of the morbidity of radical resection has led us, like many pediatric units, to re-examine multimodal treatment strategies. This review examines the Necker experience, aiming to analyze the factors influencing our decision making for treatment. PMID- 16700313 TI - Pediatric craniopharyngioma--rationale for multimodal management: the Egyptian experience. AB - BACKGROUND: Situated in a suprasellar location, craniopharyngiomas produce devastating endocrinological manifestations, visual impairment, and raised intracranial pressure, presenting a formidable disease entity for the neurosurgeon as well as the 'craniopharyngioma team'. OBJECTIVE: To evaluate direct postoperative and long-term results based on the extent of surgical removal and administration of radiotherapy. PATIENTS AND METHODS: Sixty-two new cases of pediatric craniopharyngioma were divided into three groups according to extent of removal: A (complete), B (subtotal), and C (partial). Groups A and B receiving primary postoperative radiotherapy were tagged groups Ar and Br. RESULTS: Group A had significantly less recurrence rate than group B (p < 0.005); however, direct postoperative morbidity and mortality were worse (p < 0.05). Group C fared worse than the other two groups. In groups A and B with recurrence and in groups Ar and Br, radiotherapy provided long-term control. CONCLUSION: The aim at first surgery should be total removal. Subtotal removal is warranted for more resilient tumors, while partial removal should be avoided as it provides poor results. Adjuvant radiotherapy is a useful tool in providing long-term tumor control. PMID- 16700314 TI - Management of cystic craniopharyngiomas in childhood by a transsphenoidal approach. AB - The transsphenoidal approach has specific indications in the management of craniopharyngiomas. Usually, it is best reserved for patients with preferably cystic extra-arachnoid-infradiaphragmatic tumors with small suprasellar extension. Moreover, it is definitely less traumatic than transcranial approaches and it has been proven to be feasible also in paediatric patients. When possible, radical removal of these tumours must be the goal of surgery, but this attitude, which reduces but not eliminates the risk of relapse, has to be counterbalanced by heavy morbidity and even mortality, especially in children. In this view, many neurosurgeons favour a more 'conservative' approach with subtotal removal followed by radiotherapy whose dramatic efficacy on craniopharyngiomas is well known. With these premises, a transsphenoidal approach is realistically applicable to a greater number of large cystic craniopharyngiomas if the aim is not radical removal, but is to drain them into the sphenoid sinus to relieve mass effect symptoms (cystosphenoidostomy), and delay radiotherapy and its detrimental effects on visual and pituitary function, especially in younger patients, to a more suitable time after surgery. PMID- 16700315 TI - Radiation therapy in the management of craniopharyngioma: current concepts and future developments. PMID- 16700316 TI - Review of the role of radiotherapy in craniopharyngiomas: how does patient age influence management decisions? AB - Craniopharyngioma is a brain tumor that can affect children and adults. Although histologically benign, if untreated it can enlarge and lead to serious morbidity and even death, because of its close proximity to critical structures in the parasellar region, including the pituitary, hypothalamus, optic chiasm, carotid arteries and third ventricle. Surgically, two main approaches are currently pursued: macroscopic complete excision or a primarily conservative approach. Another question is the role of radiotherapy in the postoperative setting. Because the associated morbidities with aggressive surgery or radiotherapy can be particularly pronounced in children, the most appropriate management of craniopharyngiomas in this age group remains controversial. PMID- 16700317 TI - Acute and late morbidity after limited resection and focal radiation therapy in craniopharyngiomas. AB - Aggressive surgery of craniopharyngioma can cause severe, life-long hypothalamic and pituitary dysfunctions and possibly further impair visual function; conventional radiation therapy (RT) can affect intellectual functioning and cause secondary tumours. Because of the severe morbidity associated with aggressive surgery, many authors nowadays recommend a less radical approach followed by RT. This combined approach allows achieving 70-83% 10-year local control rates which are comparable to that achieved with aggressive surgery. The main morbidity of this conservative combined approach is represented by pituitary dysfunction secondary to RT, however, sparing severe hypothalamic disturbances. The interval between treatment and onset of the disorder is much longer than in the case of aggressive surgery and this can have a beneficial impact on quality of life, especially in children. This alternative therapeutic approach has become more appealing now that modern RT techniques allow safer delivery of the RT, particularly in childhood. PMID- 16700318 TI - Does chemotherapy have a role in the management of craniopharyngioma? AB - Craniopharyngiomas are classified as histologically benign tumours that are usually treated by surgery alone or in combination with radiotherapy. However, the difficulty in managing recurrent tumours and the desire to try to avoid treatment-related morbidity from both surgery and irradiation has led to exploration of the role of chemotherapy in this tumour. In the majority of cases this has involved the application of intratumoral bleomycin in cystic craniopharyngiomas. This review reports the published experience of this type of local chemotherapy, including delivery, scheduling, outcomes and toxicity. In addition, the rarely reported use of systemic chemotherapy is discussed. PMID- 16700320 TI - Secondary narcolepsy may be a causative factor of increased daytime sleepiness in obese childhood craniopharyngioma patients. AB - Prognosis in childhood craniopharyngioma survivors hinges upon late effects such as pituitary deficiency and obesity. Observations indicate that reduced physical activity and increased daytime sleepiness might be risk factors for obesity. We analyzed the degree of daytime sleepiness in 115 childhood craniopharyngioma patients (47% obese) using the Epworth Sleepiness Scale (ESS). Thirty-five (30%) displayed increased daytime sleepiness (ESS score > 10) of whom 14 were obese (26% of obese cohort). Polysomnography (PSG) and Multiple Sleep Latency Tests (MSLT) were conducted with 10 obese patients presenting increased daytime sleepiness, with only two craniopharyngioma patients revealing a sleep related breathing disorder. Four patients had repeated episodes of SOREM (sleep onset rapid eye movement), the classic PSG criterion for narcolepsy. Three patients displayed hypersomnia. All but one patient qualified as acutely obese. We speculate that secondary narcolepsy is an exacerbating condition of childhood craniopharyngioma obesity, supported by recent reports on orexin and narcolepsy which suggest hypothalamic failure in idiopathic narcolepsy. PMID- 16700319 TI - Diabetes insipidus in craniopharyngioma: postoperative management of water and electrolyte disorders. AB - Pre-operative central diabetes insipidus has been reported in 8-35% of patients affected with craniopharyngioma, and in 70-90% after surgery. The management of postoperative polyuria and polydipsia can be challenging and fluid balance needs to be closely monitored. The classical triphasic pattern of endogenous vasopressin secretion--an initial phase of symptomatic diabetes insipidus occurring 24 hours after surgery; a second phase of inappropriate vasopressin secretion potentially causing hyponatraemia; and a third phase with a return to diabetes insipidus occurring up to 2 weeks later--is often complicated by cerebral salt wasting and thirst disorders. Inadequate adrenal replacement therapy and anticonvulsant agent treatment may increase the risk of life threatening hyponatraemia in the course of desmopressin (DDAVP) treatment. Appropriate management, in order to avoid life-threatening or disabling electrolyte disturbances, requires a good grasp of the relevant pathophysiology. We review here the pathophysiology and management of the multiple fluid disorders encountered following surgery for craniopharyngiomas. PMID- 16700321 TI - Endocrine and growth features in childhood craniopharyngioma: a mono institutional study. AB - OBJECTIVE: To evaluate growth and endocrine features in children with craniopharyngioma who were treated and followed up by a single institution between 1976 and 2004. PATIENTS: The records of 32 children, 18 males and 14 females, were evaluated. The mean follow-up period was 6.3 years. RESULTS: At presentation, the most common symptoms were headache, nausea and vomiting, visual impairment, and neurological changes. Some patients presented signs or symptoms of isolated or combined endocrine disorder (five polyuria and polydipsia, five growth failure, two precocious puberty, eight obesity or overweight). After tumour treatment, multiple pituitary hormonal deficiencies, especially growth hormone (GH) deficit (GHD) were found and required hormonal replacement therapy. Eight children grew normally without GH despite GHD. Hypothalamic involvement was observed in ten patients; obesity was frequent and was often associated with hyperinsulinism and hyperphagia. CONCLUSION: Anterior and posterior pituitary deficiencies following surgery are present in all patients. The growth pattern is heterogeneous. PMID- 16700322 TI - Craniopharyngioma radiotherapy: endocrine and cognitive effects. AB - Potential existing and treatment associated deficits in neurological, endocrine and cognitive function influence treatment decisions and the use of radiation therapy in children with craniopharyngioma. Neurological deficits are uncommon after radiation therapy, endocrine deficiencies are typically present prior to treatment, and cognitive effects depend on a wide range of clinical and treatment related factors in addition to radiation dosimetry. Early and accurate evaluation of these functions should be considered for all patients to provide parents and caregivers with the information necessary to plan intervention and mitigate the consequences of tumor and treatment. PMID- 16700323 TI - A multimodality approach to the treatment of craniopharyngiomas avoiding hypothalamic morbidity: a UK perspective. AB - The management of craniopharyngiomas is complex and controversial. The perception that they are benign tumours cured by radical surgical resection is not borne out by their often difficult excision, propensity to recur and invade, and high late morbidity and mortality from direct brain injury. Their central location makes visual or pituitary dysfunction and/or hydrocephalus common presenting features. The most important consequence of craniopharyngiomas is hypothalamic injury which may result in severe, crippling and life-threatening sequelae, such as adipsia, morbid obesity, sleep, and behavioural and cognitive disorders. Evidence suggests that unless the tumour is smaller than 2-4 cm in the mid-line and completely resectable without additional hypothalamic or visual compromise, the surgical approach should be conservative even if this leaves residual tumour. Adjuvant focal radiotherapy to residual or recurrent disease has proven efficacy in long term tumour control. Thus concerns regarding the potential late toxicity of radiotherapy to the developing brain need to be balanced against the recognised morbidity and mortality of recurrent tumour and repeated neurosurgical interventions. Hypothalamic damage is in many cases surgically induced. Given the devastating consequences of such an injury this should be avoided at all costs. Thus the aims of primary treatment should be to relieve tumour-associated compression symptoms, preserve (or improve) vision and hypothalamo-pituitary function, and minimise tumour recurrence with its attendant high morbidity and mortality. National registration and management by multidisciplinary teams in specialised centres according to nationally agreed risk adapted treatment strategies are likely to improve outcomes, as has been demonstrated for example in adult pituitary tumours. Because of the rarity of craniopharyngiomas, international collaborative trials are necessary to properly inform future therapies. PMID- 16700324 TI - Consensus and perspectives on treatment strategies in childhood craniopharyngioma: results of a meeting of the Craniopharyngioma Study Group (SIOP), Genova, 2004. PMID- 16700325 TI - Genetic modification of hepatocytes towards hepatocyte transplantation and liver tissue engineering. AB - Cell-based therapies, including liver tissue engineering following hepatocyte transplantation, have therapeutic potential for several types of liver diseases. Modifications in the methodology to manipulate the donor hepatocytes in a more simple and timely manner prior to transplantation would enhance the therapeutic efficacy of this procedure. Conventional approach for vector-mediated gene transduction to the isolated hepatocytes has been performed under primary culture conditions that routinely require several days to complete. In our study, we have established a clinically feasible approach that requires only 1 h of infection time with an adenoviral vector system that results in an extremely efficient transduction efficiency (> 80%). To optimize transduction efficiency and sustain normal cellular function, we determined that the isolated hepatocytes should be maintained in UW solution as a suspension medium and infected with adenoviral vectors (Ad) for no more than 1 h at a MOI of 1. To establish if the isolated hepatocytes could be used as a source for cell-based therapies, we transplanted the Ad-transduced hepatocytes into the liver or under the kidney capsule. When the cells were transplanted into the liver, Ad-transduced hepatocytes cultured in suspension conditions were found to have a significantly higher survival rate (p < 0.01) than Ad-transduced hepatocytes cultured under standard conditions. We also confirmed that these Ad-transduced hepatocytes have ability to survive long term and were able to engineer a biologically active hepatic tissue under the kidney capsule. Finally, we obtained high level of transduction into canine, porcine, and human isolated hepatocytes in a suspension solution mixed with Ad. In conclusion, the present studies demonstrate that isolated hepatocytes could be genetically modified using Ad when kept in a suspension solution. For this reason, this cell-modified technique could be used for the treatment of liver targeted diseases and/or disorders. PMID- 16700326 TI - Hepatocyte transplantation in the Long Evans Cinnamon rat model of Wilson's disease. AB - Wilson's disease (WD), caused by a mutation in the P-type copper transporting ATPase (Atp7b) gene, results in excessive accumulation of copper in the liver. Long Evans Cinnamon rats (LEC) bear a mutation in the atp7b gene and share clinical characteristics of human WD. To explore hepatocyte transplantation (HT) as therapy for metabolic liver diseases, 8-week-old LEC rats (n = 12) were transplanted by intrasplenic injection of hepatocytes from donor Long Evans (LE) rats. Immunosuppression was maintained with intraperitoneal tacrolimus. The success of HT was monitored at 24 weeks of life. Serum aminotransferases and bilirubin peaked at 14-21 weeks in both HT rats and nontransplanted controls, but at 24 weeks, survival was 97% in LEC-HT versus 63% in controls. All transplanted rats showed restored biliary copper excretion and reduced liver iron concentration associated with increased ceruloplasmin oxidase activity. Liver tissue expressed atp7b mRNA (11.9 +/- 13.6%) indicative of engraftment of normal cells in 7 of 12 HT rats, associated with a reduced liver copper concentration compared to untreated LEC rats. Periportal islets of normal appearing hepatocytes, recognized by atp7b antibody, were observed in transplanted livers while lobular host cells showed persistent pleomorphic changes and inflammatory infiltrates. In conclusion, transplantation of normal hepatocytes prevented fulminant hepatitis, reduces chronic inflammation, and improved 6-month survival in LEC rats. Engraftment of transplanted cells, which express atp7b mRNA, repopulated the recipient liver with normal functional capacity. PMID- 16700327 TI - Cryopreserved fetal liver cell transplants support the chronic failing liver in rats with CCl4-induced cirrhosis. AB - Hepatocyte transplantation is a promising method for supporting hepatic function in a broad spectrum of liver diseases. The aim of this work was to test the efficacy of human fetal liver cells to support the chronic failing liver in an experimental model of carbon tetrachloride (CCl4)-induced cirrhosis in rats. Liver cirrhosis was induced by intraperitoneal administration of CCl4 at a dose of 0.2 ml (50% v/v solution)/100 g body weight, twice a week for 3 months in rats. Ten days after stopping CCl4 administration (experimental day 0), rats received intrasplenic injection of cryopreserved fetal liver cells (FLC, 1 x 10(7) cells in 0.3 ml medium). As a cirrhotic control group, CCl4-induced cirrhotic rats were used with intrasplenic injection of an equal volume of medium alone. Animals were sacrificed on experimental day 15. Human fetal liver cell transplantation almost completely prevented the death of cirrhotic animals during the 2 weeks after treatment, while high ongoing mortality was seen in the cirrhotic control group. Cell transplantation into the spleen normalized total bilirubin and TBARSs levels and increased albumin levels in blood serum, as well as restoring mitochondrial function and liver detoxification function (assessed by cytochrome P450 contents and activity) compared with the activities seen in the cirrhosis control group. In parallel with this restoration of biochemical and functional liver indices, morphological patterns of liver recovery or regeneration after liver cell transplantation were demonstrated in day 15 samples by light microscopy. These were absent in the group that had received only medium alone. PMID- 16700328 TI - Analysis of the effects of cryopreservation on rat hepatocytes using SELDI-TOF mass spectrometry. AB - Successful cryopreservation of hepatocytes is essential to the future of hepatocyte transplantation as a treatment for liver disease, and also for the improved in vitro use of hepatocytes for research. However, hepatocyte function is adversely affected by even the best cryopreservation protocols. To investigate possible mechanisms for these changes, total cell lysates were prepared from fresh and cryopreserved rat hepatocytes and the proteome profiles compared using SELDI-TOF-MS ProteinChip technology. In addition, in vitro functional assays (viability, attachment efficiency, and lactate dehydrogenase leakage) were performed on the corresponding fresh and cryopreserved hepatocytes. Sixty-one peptides were identified as being significantly changed after cryopreservation. Thirty-seven peaks were significantly increased and 24 were significantly decreased after cryopreservation. The peak intensity of a number of these peptides was found to correlate with the in vitro function of the hepatocytes. Seven peptides correlated with in vitro function after cryopreservation and 10 peptides correlated with both fresh and cryopreserved function. The peptides significantly decreased after cryopreservation could include cytosolic enzymes or cofactors, which leaked out of the cells due to cryopreservation-induced membrane damage. The peptides significantly increased after cryopreservation could be retained products of cleavage of larger intracellular polypeptides and proteins or the result of aggregation of peptides caused by physical changes in the cell due to the cryopreservation process. Proteome profiling using SELDI-TOF-MS could be a useful tool to assess the effects of isolation and cryopreservation of hepatocytes, particularly if the findings are extended to human hepatocytes. PMID- 16700329 TI - Sertoli cells induce systemic donor-specific tolerance in xenogenic transplantation model. AB - Cell therapy is a potentially powerful tool in the treatment of many grave disorders including leukemia, immune deficiencies, autoimmune diseases, and diabetes. However, finding matched donors is challenging and recipients may suffer from the severe complications of systemic immune suppression. Sertoli cells, when cotransplanted with both allo- and xenograft tissues, promote graft acceptance in the absence of systemic immunosuppression. How Sertoli cells do this is not, as yet, clearly defined. We have examined the ability of Sertoli cells to produce systemic immune tolerance. For this purpose, Sertoli cells were injected into an otherwise normal C57/BL6 mouse host via the lateral tail vein. No other immunosuppressive protocols were applied. Six to 8 weeks posttransplantation, blood was collected for analysis of cytokine levels. Tolerance to donor cells was determined by mixed lymphocytic culture, and production of T-cell-dependent antibody was determined by an in vitro anti-sheep red blood cell plaque-forming assay. Results showed a marked modulation of immune cytokines in the transplanted mouse host and donor-specific transplantation tolerance was achieved. Tolerant mouse lymphocytes maintained a competent humoral antibody response. Additionally, C57/BL6 mice transplanted with rat Sertoli cells tolerated rat skin grafts significantly longer than control non-Sertoli cell transplanted mice. We conclude that systemic administration of rat Sertoli cells across xenogenic barrier induces transplantation tolerance without altering systemic immune competence. These data suggest that Sertoli cells may be used as a novel and potentially powerful tool in cell transplantation therapy. PMID- 16700330 TI - Effects of simulated microgravity on the morphology and function of neonatal porcine cell clusters cultured with and without Sertoli cells. AB - Human islet allografts are well known to induce full and sustained remission of hyperglycemia, with complete normalization of key metabolic parameters. Nevertheless, acquiring human islets, even from cadaveric human donor pancreases, remains a significant impediment to successful transplantation therapy for diabetes. To overcome this difficulty, neonatal porcine cell clusters (NPCCs) have been considered for human islet substitutes because they are easily obtained by collagenase digestion of the neonatal piglet pancreas. Currently, the major hurdle in using NPCCs for xenograft is the delay (time lag) in achieving the posttransplant normalization of blood glucose levels in animal diabetic recipients. The present work is the first attempt to evaluate whether incubation of NPCCs in simulated microgravity, in the presence or absence of Sertoli cells (SC), may reduce the maturation time lag of beta-cells by differentiation acceleration in vitro, thereby expediting production, viability, and acquisition of functional competence of pretransplantation beta-cell-enriched islets. Following a 3-day incubation period, NPCCs maintained in conventional culture, NPCCs incubated in simulated microgravity in the HARV biochamber, and NPCCs plus co-incubated SC in simulated microgravity were examined for viability, morphology, and insulin secretion. Results show that NPCCs grown alone in the HARV biochamber are superior in quality, both in terms of viability and functional competence, when compared to other culture pretreatment protocols. This finding strongly suggests that NPCC pretreatment in simulated microgravity may enhance the transplantation success of NPCCs in the diabetic recipient. PMID- 16700331 TI - Engineered cell therapy for sustained local myocardial delivery of nonsecreted proteins. AB - Novel strategies for the treatment of congestive heart failure have taken the form of gene and cell therapy to induce angiogenesis, optimize calcium handling by cardiac myocytes, or regenerate damaged myocardial tissue. Arguably both gene- and cell-based therapies would be benefited by having the ability to locally deliver specific transcription factors and other usually nonsecreted proteins to cells in the surrounding myocardial tissue. The herpes simplex virus type 1 (HSV 1) tegument protein VP22 has been shown to mediate protein intercellular trafficking to mammalian cells and finally localize into the nucleus, which makes it a useful cargo-carrying functional protein in cell-based gene therapy. While VP22 has been studied as a means to modulate tumor growth, little is known about the distribution and transport kinetics of VP22 in the heart and its potential application in combination with autologous cell transplantation for the delivery of proteins to myocardial tissue. The aim of this study was to evaluate the efficacy of VP22 fusion protein intercellular trafficking combined with autologous cell transplantation in the heart. In an in vitro study untransfected rat heart cells were cocultured with stably transfected rat cardiac fibroblasts (RCF) with fusion constructs of VP22. The control experiment was untransfected rat heart cells co-plated with RCF stably transfected with enhanced green fluorescence protein (eGFP). The Lewis rat model was selected for in vivo study. In the in vitro studies there was a 14-fold increase in the number of GFP positive cells 48 h after initiating coculture with VP22-eGFP RCF compared to eGFP RCF. In the rat model, transplantation of VP22-eGFP expressing RCF led to VP22-eGFP fusion protein delivery to an area of myocardial tissue that was 20 fold greater than that observed when eGFP RCF were transplanted. This area appeared to reach a steady state between 7 and 10 days after transplantation. The VP22-eGFP area consisted of eGFP-positive endothelium, smooth muscle cells, and cardiac myocytes with delivery to an area of approximately 1 mm2 of myocardial tissue. Our data suggest a viable strategy for the delivery of proteins that are not naturally secreted or internalized, and provide the first insight into the feasibility and effectiveness of cell-penetrating proteins combined with cell transplantation in the heart. PMID- 16700333 TI - Surgical care improvement project partnership launched. PMID- 16700332 TI - Molecular imaging reveals skeletal engraftment sites of transplanted bone marrow cells. AB - Molecular imaging holds great promise for the in vivo study of cell therapy. Our hypothesis was that multimodality molecular imaging can identify the initial skeletal engraftment sites post-bone marrow cell transplantation. Utilizing a standard mouse model of bone marrow (BM) transplantation, we introduced a combined bioluminescence (BLI) and positron emission tomography (PET) imaging reporter gene into mouse bone marrow cells. Bioluminescence imaging was used for monitoring serially the early in vivo BM cell engraftment/expansion every 24 h. Significant cell engraftment/expansion was noted by greatly increased bioluminescence about 1 week posttransplant. Then PET was applied to acquire three-dimensional images of the whole-body in vivo biodistribution of the transplanted cells. To localize cells in the skeleton, PET was followed by computed tomography (CT). Co-registration of PET and CT mapped the sites of BM engraftment. Multiple, discrete BM cell engraftment sites were observed. Taken together, this multimodality approach may be useful for further in vivo characterization of various therapeutic cell types. PMID- 16700334 TI - Corrections to "Joint Commission Accreditation Process" chapter in 2006 Comprehensive Manuals. PMID- 16700335 TI - The Joint Commission and the National Quality Forum announce the 2005 John M. Eisenberg Patient Safety and Quality Awards. PMID- 16700336 TI - Paying the price for war. PMID- 16700337 TI - A presumption of illness. PMID- 16700338 TI - Tailhook's legacy. PMID- 16700339 TI - Miles to go. PMID- 16700340 TI - Be prepared. PMID- 16700341 TI - Homecoming help. Teaching soldiers to be citizens. PMID- 16700342 TI - A strong public health system is crucial. PMID- 16700343 TI - The play's the thing--to fight obesity. PMID- 16700344 TI - Combat surgeon. PMID- 16700345 TI - Rebuilding bodies and mending minds. PMID- 16700346 TI - Serving the next greatest generation. PMID- 16700347 TI - Emergency management of sickle cell disease complications: review and practice guidelines. AB - The prevalence of sickle cell disease (SCD) in Minnesota is increasing because of the influx of immigrants from Africa, India, and countries in Latin America. Clinicians, families, and individuals with the disease have expressed the need to educate health care professionals in emergency settings about the standards of care for treating pain and other complications of SCD. Late last year, the Minnesota Department of Health and the Minnesota Hemoglobinopathy Collaborative created guidelines for treating patients with complications of SCD in the emergency department. This article provides additional background information to support the guidelines. PMID- 16700348 TI - Minnesota sickle cell disease emergency guidelines. PMID- 16700349 TI - Meeting an ever-changing public health agenda. The MMA Alliance has promoted good health in Minnesota for decades. PMID- 16700350 TI - Hope. PMID- 16700351 TI - Use of cold-adapted live attenuated influenza vaccine in pediatric patients. PMID- 16700352 TI - Idiopathic pure red cell aplasia: case report with review of literature. AB - Pure red cell aplasia (PRCA) is characterized by near absence of red blood cell precursors in the bone marrow with associated anemia and reticulocytopenia. Megakaryocytes and white blood cell precursors remain intact. Acute self-limited PRCA is the most common form. Acquired chronic PRCA is often idiopathic but can be associated with underlying disorders such as thymoma or autoimmune diseases. Congenital PRCA is associated with physical abnormalities. Pure red blood cell aplasia should always be included in the differential diagnosis of chronic unexplained anemia. We report a case of idiopathic pure red blood cell aplasia in an elderly male who has had a chronic anemia with recurrent blood transfusions for about 30 years. PMID- 16700353 TI - Oncology nurses share their experiences with comprehensive cancer control programs. PMID- 16700354 TI - Take five steps to plan an evidence-based comprehensive cancer control program. PMID- 16700355 TI - Electronic medical records mark a landmark shift in record keeping. PMID- 16700356 TI - [Avian influenza]. PMID- 16700357 TI - [Anatomy-physiology of the eye]. PMID- 16700358 TI - [Treating myopia by surgery or laser]. PMID- 16700359 TI - [What is orthoptics?]. PMID- 16700360 TI - [Cataract]. PMID- 16700361 TI - [Glaucoma]. PMID- 16700363 TI - [Developing and valuing the competences of nurses working in long-term care]. PMID- 16700362 TI - [Medical thermometers]. PMID- 16700364 TI - [Gastric intubation]. PMID- 16700365 TI - [Current practice of scanning examinations]. PMID- 16700366 TI - [Timolol eye drops]. PMID- 16700367 TI - Maternal health in the eastern Mediterranean region of the World Health Organization. AB - Globally, progress in improving the survival and well-being of mothers has been slow, and this holds true for the Eastern Mediterranean Region. Thus at the current pace, it is unlikely that the Millennium Development Goal related improving maternal health will be achieved in the Region by the year 2015. This paper outlines the factors contributing to maternal mortality and poor health in the Region and the challenges faced. The action needed to redress the situation and improve maternal health, and hence achieve the fourth Millennium Development Goal, is presented. PMID- 16700368 TI - Reducing maternal mortality in the eastern Mediterranean region. AB - Current efforts in some countries of the Eastern Mediterranean Region are still insufficient to achieve the fifth Millennium Development Goal on improving maternal health. Strong commitment, intensive efforts and effective national policies and strategies are now urgently required in order to translate vision into action. Such efforts and plans should target the strengthening of health systems, the expansion in the coverage of effective integrated interventions, and the recognition of the essential role of individuals, families and communities in making pregnancy safer. This article provides a background on the current situation of maternal health in the Eastern Mediterranean Region, including underlying causes and contributing factors, and describes strategic directions aimed at accelerating the reduction of maternal mortality in the Region and moving closer to the achievement of the Millennium Development Goals. PMID- 16700369 TI - Contraception use and probability of continuation: community-based survey of women in southern Jordan. AB - A community-based survey was conducted in October 2003 to investigate the determinants of contraception use and probability of 2 years continuation among ever married women of reproductive age in Karak, south Jordan. Of the 1109 participants, 61% were ever users of contraceptives in 1389 segments, median duration 24.0 months. The cumulative proportion of continuation was 92% at 6 months, 65% at 12 months and 42% at 24 months. Older age, longer duration of marriage, large number of surviving children and use of the intrauterine device independently predicted a longer duration of contraception use. Pregnancy planning (74%) was the most frequently stated reason for discontinuation. Family planning programmes should focus on reducing discontinuation and recommending methods with a higher probability of continuation. PMID- 16700370 TI - Duration and determinants of interbirth interval: community-based survey of women in southern Jordan. AB - The duration and determinants of interbirth intervals among women of reproductive age in Karak, Jordan were examined in October 2003. A multistage sampling technique was used to select 1109 ever-married women aged 15-49 years who contributed to 4349 interbirth intervals. Data were obtained by interview questionnaire and analysed with life table, Kaplan-Meier survival and Cox regression analyses. The median interbirth interval was 27.40 months. Longer interbirth interval was independently predicted by breastfeeding > or = 12 months, modern contraceptive use and pregnancy wastage; by more surviving children, presence of boys only or both boys and girls at the interval onset; by woman's higher education, older age and longer marriage; and by ideal spacing conforming with family planning norms. Concerted efforts to encourage modern contraceptive use, extend breastfeeding, promote small family size, address gender preferences and reinforce the minimum age at marriage should be made. PMID- 16700371 TI - [Knowledge, attitudes and practices about contraception in an urban population]. AB - We studied knowledge, attitudes and practices (KAP) regarding contraception of 1003 married women (aged 15-49 years) randomly recruited from private clinics (490 women) and community clinics (512 women) in Beirut in the year 2000. A standardized KAP questionnaire was administered to the 2 groups and differences between the groups were analysed. Contraceptive methods were used by two-thirds of the women. The private clinics group reported a significantly higher socioeconomic status and educational level than the community clinics group. Despite the free availability of contraceptive methods in the community clinics, the women's level of knowledge of, attitude towards and practice of contraception was significantly lower than those of women in the private clinics. Knowledge, attitudes and practices toward contraception did not depend on the free availability of contraceptive methods but rather on the socioeconomic and educational level of the women. PMID- 16700372 TI - Couple communication in family planning decision-making in Zahedan, Islamic Republic of Iran. AB - A qualitative study in Zahedan, Islamic Republic of Iran, used focus group discussions with 120 people in 8 main groups to understand more about couple's decision-making and the role of men in family planning. The study included men and women from Sunni and Shia sects as well as theologians from both sects. Shiites, lay people and theologians, believed that both husband and wife play a major role in decision-making about family planning, while Sunnis believed that men are the main decision-makers. All participants believed that men have an important role in family planning, but also that men do not like attending family planning clinics. Religious tenets were important in couple's decision-making about using contraceptives but there were confusions over different teachings. PMID- 16700373 TI - Study of unmet need for family planning in Dar Assalam, Sudan 2001. AB - In this descriptive study, the main objective was to determine the magnitude of unmet need for family planning among women of child-bearing age (15-49 years) in Dar Assalam. Using a detailed questionnaire, we studied 530 ever-married women selected randomly through a multistage sampling technique. Current use of contraception was 21.3%. Using the Westoff model, unmet need was 30.7%. Contraceptive pills were the most commonly used method, 92%. Knowledge on contraception was 61.3% for a single method and 23.4% > or = 2 methods. Knowledge was obtained mainly from relatives, 48.6%. PMID- 16700374 TI - Evaluation of antiepileptic drugs in pregnancy in a Jordanian army hospital. AB - We aimed to determine a treatment protocol for pregnant women with epilepsy. We selected 50 pregnant women from the antenatal clinic, King Hussein Medical Centre, who had been treated for epilepsy prior to pregnancy. They were divided into 3 groups. Group A (n = 16) received monotherapy with carbamazepine, group B (n = 16) received combined therapy with carbamazepine and phenytoin, and group C (n = 18) received no drugs because they refused treatment for fear of harming the fetus. Only 1 woman on monotherapy had seizures. In group B, no one had seizures, but 2 pregnancies were terminated because of neural tube defects. In group C, 5 patients had 1-2 seizures. No babies delivered to women in group C had congenital anomalies but 25% of babies born to mothers in groups A and B had minor congenital anomalies, a statistically significant difference. PMID- 16700375 TI - Malaria in late pregnancy in Al Hodeidah Governorate, Yemen. AB - To investigate the consequences of maternal malaria during late pregnancy, we conducted a preliminary study in the 2 hospitals in Al Hodeidah on 276 women who had uncomplicated vaginal deliveries. Only 17 women had malaria (Plasmodium falciparum), 6 with peripheral parasitaemia and 11 with both peripheral parasitaemia and placental malaria. Coincident infection carried a 9.44 times higher risk of preterm delivery and a 12.2 times greater risk of low birth weight (< 2500 g). Anaemia was diagnosed in 46.4%, associated with malaria in 11.7% of cases. All risk factors, rural residence (OR 5.18), maternal age < 20 years (OR 4.93) and primigravidae (OR 8.29), were significantly associated with malaria infection. PMID- 16700376 TI - Is unintended pregnancy a risk factor for depression in Iranian women? AB - A study was carried out to determine the association between unintended pregnancy and pre- and postpartum depression in 163 women in Kermanshah city, Islamic Republic of Iran. Using the self-administered Beck Depression Inventory for educated women and a special interview for illiterate women, 105 intended and 58 unintended pregnancies were studied. Of the 58 women with unintended pregnancy, 43% reported attempting to abort the fetus. Depression at 37 weeks' gestation was slightly higher in the unintended than the intended pregnancy group (53.4% versus 41.0%; relative risk = 1.3) and depression 10 days postpartum was much higher in the unintended group (48.7% versus 25.6%; relative risk = 1.9). Screening for depression is recommended for pregnant women with unintended pregnancy. PMID- 16700377 TI - Toxoplasmosis: the innocent suspect of pregnancy wastage in Duhok, Iraq. AB - To identify the true contribution of toxoplasmosis to fetal loss and bad obstetric history, we tested 310 women, 77.4% of whom had had single or multiple fetal loss, for evidence of infection. The study was conducted in Duhok, northern Iraq, from July 2002 till September 2003. All the women were examined for the presence of toxoplasma-specific IgM antibodies by enzyme-linked immunofluorescent assay; only 3 (0.97%) tested positive. We also tested 187 of the women by latex agglutination test; 55 tested positive. Histopathological examination was done for 9 pregnant women who tested positive by the latex agglutination test but we found no evidence of toxoplasma infection. The results indicate that the contribution of toxoplasmosis to fetal loss in our region is greatly overestimated. PMID- 16700378 TI - Value and pregnancy outcome of external cephalic version. AB - To determine the efficacy and pregnancy outcome of external cephalic version at > or = 37 weeks gestation, 90 women with a singleton breech fetus in King Hussein Medical Centre who had the procedure were compared with 102 women in a control group in whom the procedure was not attempted. External cephalic version was successfully performed on 59 fetuses (64%). Caesarean section was performed on 39% of patients who underwent the procedure compared to 61% of the control group. Version was more successful in multiparous than nulliparous women. External cephalic version was effective in reducing the number of caesarean deliveries in term breech infant in different obstetric settings, with no major neonatal adverse outcome. PMID- 16700379 TI - Serosurvey of hepatitis B surface antigen in pregnant Saudi women. AB - In 1990, Saudi Arabia began vaccinating all children at school entry against hepatitis B. We evaluated hepatitis B surface antigen (HBsAg) prevalence rate among pregnant Saudi women 12 years later in 5 regions of the country. Using multistage sampling, 2664 pregnant Saudi women were recruited. Blood samples were tested for HBsAg; positive samples were also tested for hepatitis Be antigen (HBeAg). In all 2.44% were positive for HBsAg and 4 (0.15%) were also positive for HBeAg. HBsAg prevalence was highest in Gizan (4.2%) and lowest in Tabuk (1.4%). Positivity for women < or = 20 years of age was 0.5% compared with 2.6% for older women (P = 0.049). The overall HBsAg prevalence rate was lower than previously reported. PMID- 16700381 TI - Termination of second trimester, complicated gestation. AB - To assess the effectiveness of intravaginal misoprostol for second trimester uterine evacuation, we studied 70 women with singleton pregnancies complicated by fetal malformation or dead fetuses. Participants received 200 microg misoprostol administered at 4-hour intervals. Gestations with dead fetuses had a shorter induction-abortion interval [14.2 hours, standard deviation (SD) 4.3] than those with live, malformed fetuses (20.2 hours, SD 7.3) (P< 0.001). The abortion rate was significantly higher for gestations with dead fetuses (92.1%) than those with live, malformed fetuses (68.8%) (P< 0.05). There were no major complications and no significant difference in the incidence of side-effects. All women aborted within 38 hours. Administration of misoprostol is an effective clinical method to terminate second trimester, complicated pregnancy. PMID- 16700380 TI - Is vitamin A deficiency a problem among pregnant Emirati women in Al-Ain City? AB - Vitamin A deficiency (VAD) can have a negative impact on pregnancy but there have been no studies in Al-Ain on the vitamin A status of pregnant women. We studied 198 pregnant Emirati women aged 15-49 years attending antenatal clinics in Al-Ain Medical District (1999-2000) to assess the prevalence of VAD. Sociodemographic and health information about the women was collected by questionnaire and they all underwent blood and serum analysis. Of the 198 women, only 6 (3%) had vitamin A deficiency (plasma vitamin A < 20 microg/dL), indicating only a mild problem according to WHO criteria. There was no significant association between the occurrence of VAD and any of the characteristics studied. While the mean values of all the haematological indices were slightly lower in the vitamin A deficient group, this was not significant. PMID- 16700382 TI - Women's knowledge and attitude towards modes of delivery in Kerman, Islamic Republic of Iran. AB - Knowledge and attitudes about vaginal versus caesarean section delivery were assessed in 204 pregnant women attending a maternity centre in Kerman, Islamic Republic of Iran. Overall, 63.5% of women scored weak on knowledge questions. Knowledge was higher in women who had a history of miscarriage. Of the women, 96.5% and 33.0% had positive attitudes towards vaginal delivery and caesarean section respectively; 40.5% had negative attitudes about caesarean section. Women with higher parity and more previous caesarean deliveries showed positive attitudes towards vaginal delivery, as did housewives and women whose spouses were employed in education or health organizations. No significant differences were found in attitude and knowledge scores according to women's levels of education. PMID- 16700383 TI - Neonatal outcome in the United Arab Emirates: the effect of changes in resources and practices. AB - Selective improvements in neonatal care resources and practices were instituted between 1992/1994 (period 1) and 1995/1998 (period 2) following a neonatal audit in the United Arab Emirates. We evaluated the effect of these changes on neonatal mortality rate (NNMR), birth-weight-specific mortality rates and causes of mortality. Overall there was a 17% decline in the NNMR from periods 1 to 2. Mortality rates in infants with birth weight < 1000 g and > 2500 g decreased by 36% and 35% respectively from periods 1 to 2. Modest declines in deaths from asphyxia, sepsis and complications of preterm births occurred from periods 1 to 2 but the differences were not statistically significant. PMID- 16700385 TI - The National Congenital Anomalies Register in the United Arab Emirates. AB - The National Congenital Anomalies Register is a population-based register covering all births in the United Arab Emirates. We evaluated the progress of the register and determined the prevalence of congenital anomalies (CAs) and associated maternal and neonatal risk factors. Total prevalence of CAs for 1999 2001 was 7.89/1000, 10.95/1000 and 7.92/1000 for live births, stillbirths and total births respectively. Rates were comparable to international rates for all districts except Dubai, Fujairah and Ras Al Khaimah. According to the International classification of diseases, the cardiovascular system was the most affected followed by CAs of chromosomal origin and the musculo-skeletal system. Birth defects were more common with older maternal age, grand multiparity, male babies, low-birth-weight babies and premature babies. PMID- 16700384 TI - Burden of perinatal conditions in Yemen: a 12-year hospital-based study. AB - We conducted a 12-year retrospective analysis of perinatal characteristics at Al Sabeen Hospital, Sana'a. There were 62168 births, 2936 stillbirths and 5434 perinatal deaths. There were 14 576 (24.6%) preterm low-birth-weight (LBW) babies. Early neonatal death (ENND) rate was 42.2/1000 live births. Of the 10 546 neonates admitted to the special baby care unit (SBCU), 40.1% were preterm LBW and 2147 (20.4%) died (80.6% were preterm LBW). The last 4 years showed a 17.2% increase in stillbirths and a 40.5% reduction in ENND compared with the first 4 years. In the SBCU, reduction in ENND was > 70%. Respiratory distress accounted for 63.8% of deaths in the SBCU and birth asphyxia 15.0%. Our results reflect the poor antenatal and delivery care services in Yemen. PMID- 16700386 TI - Profile of major congenital malformations in neonates in Al-Jahra region of Kuwait. AB - We investigated major congenital abnormalities in babies born in Al Jahra Hospital, Kuwait from January 2000 to December 2001. Of 7739 live and still births born over this period, 97 babies had major congenital malformations (12.5/1000 births): 49 (50.6%) babies had multiple system malformations, while 48 (49.4%) had single system anomalies. Of the 49 babies with multiple malformations, 21 (42.8%) had recognized syndromes, most of which were autosomal recessive and 17 had chromosomal aberrations. Isolated systems anomalies included central nervous system (12 cases), cardiovascular system (9 cases), skeletal system (7 cases) and gastrointestinal system (6 cases). Of the parents, 68% were consanguineous. Genetic factors were implicated in 79% of cases. Genetic services need to be provided as an effective means for the prevention of these disorders. PMID- 16700387 TI - Congenital malformations at a referral hospital in Gorgan, Islamic Republic of Iran. AB - This study recorded the rate of congenital malformations in 10000 births at a referral hospital in Gorgan, Islamic Republic of Iran in 1998-99. The overall incidence of congenital malformations was 1.01% (1.19% in males and 0.76% in females). Anomalies of the musculoskeletal system had the highest incidence (0.38%), followed by central nervous system (0.28%) and genitourinary system (0.25%). The incidence of congenital malformations in different ethnic groups was 0.85%, 1.45% and 1.70% in native Fars, Turkman and Sistani groups respectively. Sex and ethnic background are factors in the rate of congenital malformations in this area. PMID- 16700388 TI - Effects of antenatal steroids on the incidence and severity of respiratory distress syndrome in an Iranian hospital. AB - Antenatal corticosteroids are still not routinely used in the Islamic Republic of Iran to promote lung maturity in premature neonates. In a retrospective review of records at Imam Hospital, Tehran, we compared 140 premature neonates born in 2000 whose mothers received dexamethasone antenatally with 142 born in 1995 whose mothers did not receive treatment. There were no significant differences between the groups in mothers' age; neonates' birth weight, gestational age or sex; route of delivery and multiple gestation; underlying maternal disease; or adverse events in labour. The incidence of respiratory distress syndrome was significantly lower in the dexamethasone-treated group (18.6% versus 35.9%). Mortality (5.7% versus 14.8%) and use of the neonatal intensive care unit (12.9% versus 21.1%) were also significantly lower among those who had received treatment. PMID- 16700389 TI - Morbidity and mortality of low-birth-weight infants in Egypt. AB - Low birth weight is one of the major determinants of neonatal survival and postneonatal morbidity. This study assessed the morbidity and mortality of low birth-weight (LBW) infants during the first 3 months. Data were collected for 6701 live births. All singleton liveborn LBW infants as well as a comparison group of normal-birth-weight (NBW) infants were followed up for 3 months. Data were collected on weight, morbidity and mortality using a structured questionnaire. Admission to neonatal intensive care unit and mortality were more frequent in LBW (31.6%, 2.0%) than NBW infants (2.0%, 0.2%). They also had increased risk of neonatal jaundice at 1 month, an increased risk of growth retardation and a much higher risk of mortality. PMID- 16700390 TI - Frequency of visits for health care by low-birth-weight and preterm Saudi infants in the first year of life. AB - A retrospective hospital-based cross-sectional survey in a Saudi Arabian hospital compared the frequency of visits to hospital of low-birth-weight and normal-birth weight infants and of preterm and full-term infants during the first year of life. A multivariate analysis of the visits for health care with birth weight, term, Apgar score, mode of delivery and sex was made for 1892 infants. The number of visits for health care was significantly higher for low-birth-weight and preterm infants. While the mode of delivery had no effect on the frequency of visits, male sex and poor Apgar score 5 minutes after birth were associated with greater frequency of visits to specialty clinics and higher rates of hospitalization. PMID- 16700392 TI - Clinical and laboratory aspects of malaria among children with fever in a low transmission area of Sudan. AB - A cross-sectional study explored the clinical and laboratory aspects of malaria among children presenting with fever to 2 paediatric hospitals in Khartoum state during the low transmission season. Out of 655 febrile patients, 35.9% were recorded as having malaria based on hospital laboratory results. However, re examination of slides at the National Malaria Control Programme referral laboratory confirmed malaria in only 32.8% of those diagnosed with malaria at hospital level. Analysis of symptoms and signs revealed great variability in clinical presentation. Although some findings were associated with malaria, developing a sensitive clinical algorithm was difficult. Further investment is needed to improve microscopic diagnosis facilities in local hospitals to overcome the problem of over-diagnosis of malaria. PMID- 16700391 TI - Effects of extra immunization efforts on routine immunization at district level in Pakistan. AB - A study was made of the effects of extra immunization activities on routine immunization coverage at district level in Pakistan in a secondary analysis of data from UNICEF and the Pakistan national census. Linear regression analysis was made on data from 107 districts to estimate the effects of extra immunization efforts in the national neonatal tetanus programme on the coverage rate of the third dose of diphtheria-pertussis-tetanus (DPT3) after controlling for other potential confounding factors. The districts that implemented extra national neonatal tetanus immunization were at risk of having lower routine DPT3 coverage than those that did not. Additional immunization efforts, without additional resources, may reduce the effect of the routine Expanded Programme on Immunization. PMID- 16700393 TI - Epidemiology of diarrhoeal diseases among children under age 5 years in Dakahlia, Egypt. AB - Using multistage cluster sampling, we conducted a household survey of diarrhoea among 4458 children under age 5 years in Dakahlia governorate from June 2002 to May 2003 to determine the prevalence and determinants of diarrhoeal diseases. Frequency of diarrhoea in the previous 2 weeks and last 24 hours were 23.6% and 8.7% respectively. Oral rehydration solution use rate was 24.3% among children with diarrhoea in the past 2 weeks. The frequency of diarrhoea was significantly higher among children in rural areas, those aged 6-24 months and of higher birth order, in the summer, when mothers were younger, had lower education or were not working, and when fathers had lower education or were farmers or manual labourers. Overcrowding, improper refuse disposal and non-flush toilets were also significantly correlated with diarrhoea incidence. PMID- 16700394 TI - [Swimming programme for mentally retarded children and its impact on skills development]. AB - People with Down syndrome suffer from many social, psychological and motor function problems. Current research has looked at swimming programmes to assess their impact on such problems. The programme described here was applied for a 3 month period, with music being used as one of the tools. The results were very positive and children showed significant improvement in agility, self-esteem and behaviour patterns. The programme is thus effective in enhancing the integrated development of children with Down syndrome. PMID- 16700395 TI - Type 2 diabetes mellitus among children and adolescents in Al-Ain: a case series. AB - To characterize the features of type 2 diabetes mellitus among children and adolescents in Al-Ain, the records of every child with diabetes attending a teaching hospital in the city from January 1990 to December 2001 were retrospectively examined. Of 96 young people newly diagnosed with diabetes mellitus, 11 were identified as type 2. The clinical characteristics were: pubertal onset, female preponderance, obesity, strong family history of type 2 diabetes mellitus, high plasma glucose at presentation, adequate beta cell reserve and serum pancreatic islet cell antibody negativity. This case series adds to the evidence that type 2 diabetes mellitus is emerging among children in our region. PMID- 16700396 TI - Incidence of traumatic injection neuropathy among children in Pakistan. AB - We used acute flaccid paralysis surveillance data collected between 1 January 2001 and 31 December 2003 from the Pakistan Polio Eradication Initiative to describe the epidemiological characteristics and disease burden of traumatic injection neuropathy among children aged under 15 years. Of the 5627 acute flaccid paralysis cases reported, 456 were identified as traumatic injection neuropathy by case review. The condition was more common in younger children who were also more likely to have persistent paralysis. We estimate that the annual incidence of traumatic injection neuropathy rate in Pakistan is 7.1 per 1 000 000 in children under 3 years old. PMID- 16700397 TI - Adolescents' attitudes toward gender roles and women's empowerment in Oman. AB - This study investigated the attitudes of Omani adolescents towards gender roles and women's empowerment in taking household decisions, and the determinants of their positive attitudes. A national representative secondary school-based sample of 1670 boys and 1675 girls completed a self-administrated questionnaire with 2 indices: Adolescents' Attitudes Towards Gender Roles and Adolescents' Attitudes Towards Women's Empowerment. Other questions included demographic data, risk behaviour lifestyle, proxies to mental health and social environments expected to predict attitudes towards gender role. Eight variables predicted higher scores in both indices: sex, age, socioeconomic class, sleep problems, self-esteem, current smoking, history of indulging in violent behaviour and parental relations. PMID- 16700398 TI - Incidence of neural tube defects among neonates at King Hussein Medical Centre, Jordan. AB - Neural tube defects (NTDs) are important factors in fetal and infant mortality. There have been no studies on the incidence of NTDs in Jordan. A one-year prospective study was therefore conducted in King Hussein Medical Centre neonatal unit (April 2002-April 2003) to determine the incidence of NTDs among Jordanian neonates. All liveborn babies, both term and preterm, were assessed for NTDs. Mothers of babies with NTDs were interviewed postnatally regarding antenatal care and family history of NTDs. Of 5088 live births, 33 had NTDs, giving an incidence of 6.5/1000 live births. None of the women with affected babies had a history of NTDs and all had received antenatal care, but none had taken folate supplementation. The majority of the women had not had a diagnosis before the birth, despite having had ultrasound scans throughout their pregnancies. PMID- 16700399 TI - Maternal serum screening of Palestinian women in the West Bank. AB - An audit was made of laboratory records of triple test results from 943 Palestinian women residing in the West Bank from 2000-03. The mean expected age at delivery of the subjects was 25.5 years; 12% were 35 years old or above. Age was the main factor associated with an abnormal calculated risk score. The frequency of elevated serum alpha-fetoprotein was 1.3%, abnormal human choriogonic gonadotropin (elevated or reduced) was 1.9% and reduced unconjugated estriol was 0.3%. PMID- 16700400 TI - Couples' attitudes to the husband's presence in the delivery room during childbirth. AB - We determined the attitudes of men and women in the Islamic Republic of Iran to the husband's presence in the delivery room. We randomly selected 150 couples awaiting delivery in 2002 and interviewed them with a standardized questionnaire. Mean ages and standard deviations (SD) of women and men were 26 (SD 4) and 30 (SD 5) years respectively. Most had high school diplomas or higher. Most women (58.1%) were employed and most men (77.6%) had non-educational jobs. Mean attitude scores were 100 (SD 15) for women and 97 (SD 16.5) for men. Attitude scores were significantly related to age, job and education (P< 0.0001). Most women (88.4%), men (82.1%) and couples (76.9%) had positive attitudes to the husband's presence in the delivery room. Providing facilities to accommodate husbands and training for their presence in the delivery room is recommended. PMID- 16700401 TI - Midsole material-related force control during heel-toe running. AB - The impact maximum and rearfoot eversion have been used as indicators of load on internal structures in running. The midsole hardness of a typical running shoe was varied systematically to determine the relationship between external ground reaction force (GRF), in-shoe force, and kinematic variables. Eight subjects were tested during overground running at 4 m/s. Rearfoot movement as well as in-shoe forces and external GRF varied nonsystematically with midsole hardness. Kinematic parameters such as knee flexion and foot velocity at touchdown (TD), also varied nonsystematically with altered midsole hardness. Results demonstrate that considerable variations of in-shoe loading occur that were not depicted by external GRF measurements alone. Individuals apparently use different strategies of mechanical and neuromuscular adaptation in response to footwear modifications. In conclusion, shoe design effects on impact forces or other factors relating to injuries depend on the individual and therefore cannot be generalized. PMID- 16700402 TI - The characteristics of foot movements in Tai Chi Chuan. AB - This study examines how the characteristics of the foot movements in Tai Chi Chuan (TCC) contribute to a practitioner's health by improving his or her balance and reducing the risk of fall. Ten TCC masters are video-taped, and the time spent on different support patterns (step stance) and stepping directions (footwork) while performing the 42-form TCC is analyzed. In support patterns, it is found that a larger percentage of time is spent on the double leg stance (35.08 +/- 4.92) than on the single leg stance (Left: 17.67 +/- 7.71; Right: 17.29 +/- 2.23) and one leg support with another leg partially supporting type of stance (6.51 +/- 8.77). With regard to stepping directions, the centre footwork (25.25 +/- 10.20) requires a larger percentage of time than the forward (18.29 +/ 3.36), sideway (20.80 +/- 4.76), grinding (20.42 +/- 7.61), upward (11.55 +/- 1.49), and backward footwork (3.67 +/- 1.33). The movements classified are shown to simulate balance, flexibility and proprioception, and functional training. The findings partially explain the relationship between practising TCC and its health benefits. Further studies are suggested to investigate the health benefits that can be derived from other specific TCC movements. PMID- 16700403 TI - Physiological and mechanical response to soccer-specific intermittent activity and steady-state activity. AB - The aim of this study was to quantify response to a soccer-specific intermittent (INT) treadmill protocol based on notational analysis of match-play. Ten male semiprofessional football players (age 24.7 +/- 4.4 yr, body mass 77.1 +/- 8.3 kg, VO2max 63.0 +/- 4.8 ml x kg x min(-1)) completed the 90 minute INT protocol and a steady-state (SS) protocol eliciting the same distance covered. Physiological (heart rate [HR], ratings of perceived exertion [RPE], blood lactate concentration, salivary cortisol concentration) and mechanical (electromyography [EMG] of biceps femoris and rectus femoris) responses were obtained at 15 minute intervals throughout each protocol. The physiological and mechanical responses were typically greater during the INT protocol than during the SS protocol, tending to increase as a function of exercise duration. The INT activity profile induces cumulative mechanical load on the musculoskeletal system. The increased incidence of injury toward the latter stages of match-play is attributed to compromised movement mechanics, rather than physiological strain. PMID- 16700404 TI - Effect of a liquid multivitamin/mineral supplement on anaerobic exercise performance. AB - The purpose of this study was to determine if supplementation with a liquid multi vitamin/mineral would improve anaerobic exercise performance. Fourteen resistance trained men performed a 30-second cycle sprint and one set of squat exercise on 2 separate days before and following 8 weeks of supplementation with either a liquid multi-vitamin/ mineral or a placebo. Heart rate, perceived exertion, blood lactate, peak and mean power, and rate of fatigue were determined for all tests. No differences were noted for any variable (P > 0.05). When controlling for presupplementation values, however, a decreased rate of fatigue was noted for both exercise tests following the multi-vitamin/mineral supplementation. These data suggest that in resistance trained men consuming a nutritionally sound diet, supplementation with a liquid multi-vitamin/mineral does not favorably impact most anaerobic exercise performances. Such supplementation, however, may result in a minor decreased rate of fatigue. It appears that, in terms of improved short duration anaerobic exercise performance, supplemental micronutrients may not be efficient ergogenic agents for well-trained individuals consuming an adequate diet. PMID- 16700405 TI - Review of exercise-induced muscle injury: relevance for athletic populations. AB - Exercise-induced skeletal muscle injury is well understood as the product of unfamiliar or strenuous physical activity. Eccentric or lengthening actions are primarily responsible for inducing injury, which subsequently leads to a variety of signs and symptoms. Although significant research supports this finding, most observations are specific to untrained individuals. In addition, many protocols designed both to induce muscle injury and assess performance following the injury are dissimilar from those utilized by physically trained individuals or are impractical in relation to athletic performance or both. Therefore, difficulty may arise when extrapolating information, from the available literature and applying the findings to athletic populations. This review addresses the efficacy of applying our current understanding of exercise-induced skeletal muscle injury to a physically trained population as well as highlights concerns that require future investigation. PMID- 16700406 TI - The healthy worker effect in major league baseball revisited. AB - We investigated the "healthy worker effect" (HWE) in major league baseball. Ages of death of major league baseball players who debuted between 1900 and 1939 were obtained and differences between those ages and age-adjusted life expectancies were examined to determine if longevity increased with career length, controlling for decade in which a player debuted, player position, and handedness. Major league baseball players (N = 4,492) lived an average of 4.8 (+/- 15.0 Standard deviation [SD]) years longer than age-matched controls from the general public. Career length significantly and incrementally increased longevity of players from an average of 4.1 years for players playing one season to 7.4 years for players playing 11 or more years. None of the other factors, nor any of the interactions, was statistically significant. These data provide strong support for the HWE in professional baseball. PMID- 16700407 TI - [Conventional surgery for congestive heart failure]. AB - Congestive heart failure is a major public health problem in western countries. Although substantial efforts have been made in the last decades in the prevention, diagnosis and treatment of cardiovascular disease, the incidence of end-stage dilated cardiomyopathy is still increasing. Heart transplantation represents the most effective therapy in this setting, but due to shortage of donors, it remains a realistic option just for a very small number of patients. Therefore, conventional surgical treatment for end-stage heart disease has gained increasing attention in recent years and a variety of surgical interventions have been improved or optimized to manage the multifactorial pathophysiology of the heart failure picture. The aim of this review is to report our experience with more than 500 patients with advanced dilated cardiomyopathy, treated with conventional surgical procedures such as myocardial revascularization, left ventricular restoration, mitral valve repair and surgical ablation of atrial fibrillation. Indications, results, controversial issues and future perspectives will be discussed. PMID- 16700408 TI - [Cardiac cell therapy: the puzzle is waiting to be solved]. AB - Cell therapy has been proposed as an innovative hypothesis to treat acute myocardial infarction and heart failure. However, the mechanism by which stem cells could improve cardiac function remains unclear and many controversies have been arisen in interpretation of experimental and clinical data. Answering the five "WH questions" we discuss the process that has led to consider cell therapy as a new treatment option for myocardial tissue regeneration after ischemic damage. 1) Why should we use stem cells? The rationale derives from the disclosure that apoptosis and regeneration occur at the myocardial level and stem cells migrate from bone marrow to repopulate the damaged cardiac tissue. 2) Which are the most appropriate cells, delivery methods and therapeutic purposes? Adult stem cells can be mobilized or directly transplanted in human hearts to accomplish myocardioneogenesis, neoangiogenesis and/or paracrine effects. 3) Where should we transplant these cells? The infarct border zone seems to be the best place to home and differentiate transplanted cells hampering post-ischemic cardiac remodeling. 4) When should we perform cell therapy? Cell therapy should be performed during or after an acute myocardial infarction: best setting and timing still need to be precisely addressed. 5) Who might be the suitable patient? Further multicenter randomized trials with adequate patient selection are needed to answer this crucial question. PMID- 16700409 TI - [Rheumatic fever and rheumatic heart disease]. AB - Rheumatic heart disease, the sequel of acute rheumatic fever, is a very common cause of cardiovascular mortality and morbidity all over the world, and is the predominant indication for cardiac surgery in the industrialized countries. Diagnosis of rheumatic chronic carditis may sometimes be difficult because valvular regurgitation may not always be detected by routine clinical auscultation. A recent report from the World Health Organization Expert Committee recognizes the usefulness of echocardiography Doppler in providing supporting evidence for diagnosis of rheumatic carditis in the presence of equivocally pathological murmur, and recommends that patients with subclinical carditis should be managed as rheumatic heart disease until proven otherwise, because the disease still represents a major health problem. The aim of this review is to give an update on the disease by underlining changes made by the World Health Organization on disease diagnosis and patient management. PMID- 16700410 TI - [Psychological characteristics of patients with vasovagal syncope: and observational study on sixty subjects]. AB - BACKGROUND: Based on the relevant literature, the aim of this study was to analyze the psychosocial characteristics of patients with vasovagal syncope and to evaluate these factors as possible etiopathological components. METHODS: The study sample consisted of 60 subjects divided as follows: the syncope group (n = 30) and the control group (n = 30, without prominent diseases). The two groups were matched with regard to age, sex, education and civil status. Each participant filled in the Twenty-Item Toronto Alexithymia Scale, acceptance of emotions scale, profile of mood state, attachment style questionnaire, and childhood traumatic events scale. RESULTS: The syncope group showed a higher number of somatic diseases (p < 0.0002) and a higher drug use (p < 0.0001) than the control group. Moreover, the syncope group showed higher scores at the childhood traumatic events scale (p < 0.04) and more difficulties in emotion regulation (p < 0.02). The syncope group also showed higher scores at the need for approval scale (p < 0.0006) and lower scores at the confidence scale (p < 0.02) of the attachment style questionnaire, which reflect an insecure style of relationship with others. CONCLUSIONS: A high number of traumatic events in infancy and adolescence, difficulties in the expression of emotions and an insecure style of relationship seem to characterize subjects with vasovagal syncope. These data show that syncope could be at least partiality due to a relational and emotional imbalance that finds expression through the body in the presence of insufficient mental processing. Treatment of this syndrome should therefore take these aspects into consideration. PMID- 16700411 TI - [Percutaneous coronary intervention and immediate re-transfer to the referring hospital for patients with acute coronary syndrome. A single-center experience]. AB - BACKGROUND: Current guidelines recommend early transfer of patients with acute coronary syndromes from primary care centers to hospitals with cath lab facilities. Few data exist about safety of re-transfer to primary care centers immediately after successful percutaneous coronary interventions. METHODS: We analyzed data regarding 102 consecutive patients (73 male, 29 female, mean age 69 +/- 10.9 years) transferred to the Bolzano hospital from centers without cath lab facilities for urgent percutaneous coronary intervention in acute coronary syndromes and planned immediate re-transfer after the procedure. Data about complications during re-transfer were obtained using a prepared data sheet completed by physicians and/or paramedics involved in the transport. RESULTS: Eighty-eight (87.1%) patients were re-transferred immediately after the cath lab procedure. Arterial hemostasis was achieved with arterial closure devices before leaving the cath lab. During re-transfer, 61 (69%) patients were accompanied by physicians and 27 (31%) patients by paramedics. No ischemic, arrhythmic or hemorrhagic complications were observed. CONCLUSIONS: In our experience immediate re-transfer to the referring hospital after successful urgent percutaneous coronary intervention for acute coronary syndrome is feasible and safe. If the patient is clinically stable after the procedure, re-transfer may not require the presence of a physician. PMID- 16700412 TI - [Echocardiographic diagnosis in cardiothoracic emergencies: who is the referee? The viewpoints of the anesthesiologist, the cardiologist, the legal doctor, and the medical director]. PMID- 16700413 TI - [Natriuretic peptides and cardiac resynchronization therapy]. PMID- 16700415 TI - [When the surgeon is ... on stage]. PMID- 16700414 TI - [Severe bradycardia in an asymptomatic young subject: is there an indication to permanent cardiac pacing?]. AB - Bradycardic effects of severe weight loss are well known like those observed in anorexia nervosa resulting from an increase in vagal tone associated with a low calorie-protein diet. We here report the case of a 19-year-old patient with asymptomatic severe bradycardia, which developed after voluntary weight loss of more than 30 kg during a few months. In absence of symptoms, ventricular arrhythmia or structural heart disease, and owing to normal ventricular function, no permanent pacemaker implantation was undertaken. After a follow-up of more than 15 years, clinical conditions are stable, the patient is asymptomatic and bradycardic, and no changes in weight are reported. PMID- 16700416 TI - [New tools for a better diagnosis of latent tuberculosis infection]. AB - Two new blood tests are today available for diagnosing latent tuberculosis infection. Both tests are based on the release of interferon-gamma from M. tuberculosis-specific T cells. These tests, QuantiFERON-TB Gold and T-SPOT.TB, are certainly more specific compared to the tuberculin skin test, and possibly more sensitive in some subgroups of patients; they might represent a crucial tool for tuberculosis control and elimination. PMID- 16700417 TI - [Illness perception questionnaire (IPQ-r): an useful paradigm in chronic disease]. AB - The Illness Perception Questionnaire (IPQr) is a validated psycho-social tool. Chronic illness out-patients, 48 women and 42 men, 61.36 +/- 14.55 yrs, showed a social stigma related to illness condition, even common and scarcely invalidating, more evident with a lower degree of instruction, associated to limited cognitive representation and to inappropriate control. Duration of illness directly influences perception of "chronicity"; age, instead, positively influences compliance and trust of patients. The study of illness perception, aimed to foresee, interprets and modifies behavioural patterns in health and disease, according to implicit and/or hidden personal and/or social theories, it can improve quality of clinical medical approach. PMID- 16700418 TI - [Correlation between different clinical activity and anti CC-P (anti-cyclic citrullinated peptide antibodies) titres in rheumatoid arthritis treated with three different tumor necrosis factors TNF-alpha blockers]. AB - TNF-alpha role in RA is confirmed by the improvement on joint inflammation and physical function and by the slowing of radiographic damage induced by TNF-alpha blockers, that also reduce Rheumatoid Factor (RF) and anti-CC-P titres. (1) To evaluate the effects of 3 TNF-alpha blockers on (a) disability (HAQ), disease activity (DAS28), acute phase reactants (ERS, CRP); (b) autoantibodies: IgM RF, anti-CCP abs. (2) To evaluate if anti-CCP abs could be useful for testing the efficacy of anti-TNF-alpha agents in the follow-up of RA patients. 34 patients with RA (25 F, 9 M; mean age: 59.1 +/- 12.1 years, mean disease duration: 9.6 +/- 2.3 years; 23/34 positive for anti-CCP abs, 19/34 for IgM RF) were enrolled: 18 received Etanercept (25 mg twice weekly subcutaneously), 8 received Infliximab (3 mg/kg intravenously every 8 weeks) and 8 Adalimumab (40 mg every 14 days). All the patients were evaluated for the above mentioned parameters at baseline (t0) and after 6 months of therapy (t6). Anti-TNF-alpha agents differently reduced HAQ and DAS28, ERS and CRP, RF and anti-CCP ab titres RA patients whose RF (14) and/or anti-CCP abs (17) titres were significantly lowered at the end of the study, at t6 presented a significant reduction in respect to t0 of VES, PCR, DAS28 and HAQ values (p < 0.05 for all comparison). This effect was not shown in patients in whose RF (5) and/or anti-CCP abs (6) titres were not reduced in respect to baseline. In RA, anti-TNF-alpha agents, especially Etanercept, reduce disability, disease activity and acute phase reactants expecially in patients showing reduction of RF and anti CC-P titres. PMID- 16700419 TI - [Avascular jaw osteonecrosis associated with bisphophonate therapy]. AB - The case of an IgAk myeloma showed the following common features with more than a hundred of patients described by the literature: osteonecrosis of the jaws, prolonged administration of bisphosphonates due to neoplastic skeletal lesions, antitumoral therapies and recent dental surgery. The antiangiogenetic effects of bisphosphonates can have an important pathogenetic role and a general dental treatment should be completed before the start of bisphosphonate administration. PMID- 16700421 TI - [Alert and alarmism: "in medio stat...virus"]. PMID- 16700420 TI - [Heart failure and cholelithiasis]. AB - We previously found an high incidence of gallstone or cholecystectomy for lithiasis in patients affected by heart failure (52.1% up to 58.6%). As in general population the incidence of this disease is up to 38%, we tried to explain our findings. PMID- 16700422 TI - [Exercise electrocardiography for diagnostic and prognostic assessment of coronary disease. Recent advances]. AB - This article focuses on recent advances on exercise electrocardiography (ECG) for the diagnostic and prognostic assessment of coronary artery disease. In particular, the pathophysiological background of comparative heart rate-adjusted ST-segment analysis during exercise and recovery phase is discussed. In addition, the results of recent studies dealing with the application of this ECG analysis modality to different clinical settings are presented. PMID- 16700423 TI - [Anemia and myocardial ischemia: relationships and interferences]. AB - Of all the pathologies that cause a critical reduction in oxygen supply to the tissues, atherosclerosis is undoubtedly the most frequent. At the coronary level, ischemia causes a critical imbalance in oxygen supply and demand to the myocardium. Protracting this condition induces necrosis since, when the heart surpasses certain limits, it is incapable of modulating its metabolism in relation to the availability of energy substrates. Recently, internists and cardiologists have been examining the role of anemia as a cause or cofactor in the development of myocardial ischemia. The drop in circulating erythrocyte mass and consequent drops in hemoglobin and hematocrit represent obstacles to oxygen transport and delivery to tissues. Tachycardia secondary to anemia leads to a shorter diastolic phase and reduction in arterial pressure, phenomena which are particularly prominent in acute anemia. Such changes can result in serious repercussions for people suffering from coronary disease, leading often to documentable myocardial damages. On the other hand, in chronic anemia the compensation phenomenom--represented by an increase in heart rate, cardiac output, ventricular volume and contractility--requires integrity of the cardiac apparatus, thus explaining the very serious repercussions of the anemic state for cardiac patients. Recently, the possibility of preventing and treating anemia induced or anemia-exacerbated ischemia has been confirmed in the literature, albeit not unequivocably, demonstrating that this topic deserves particular attention. PMID- 16700424 TI - [Psychiatry and psychiatrists in the U.S.A. cinema]. AB - United States cinema motion pictures from the beginning of 20th century to the present are characterized by massive use of sterotypes to represent psychiatrist's image, as well as psychiatric treatment and inpatients psychiatric facilities. Representation tends to undergo considerable changes between psychiatric different historical periods. Psychiatric disorders also are commonly depicted in movies, often in a not realistic way. The images of psychiatrist and mental disorders shown in movies are likely to impact on the beliefs and attitudes of people towards psychiatry. PMID- 16700425 TI - [Biotech]. PMID- 16700426 TI - [Cystic fibrosis]. PMID- 16700428 TI - Comparative evaluation of conventional and alternative methods for the removal of arsenic from contaminated groundwaters. AB - The present paper intends to summarize the recent findings regarding the development of alternative treatment methods applicable to small municipal drinking water systems. Small systems are frequently affected by the new permissible arsenic concentration rules, as imposed by several international organizations-World Health Organization, European Commission, United States Environmental Protection Agency. The innovate treatment methods reviewed are (a) adsorptive filtration, using iron oxide coated sand or polymeric materials; (b) zero-valent iron; (c) solar disinfection, solar oxidation and removal of arsenic (SORAS); (d) iron-based adsorbents, such as granular ferric hydroxide (GFH); (e) biological oxidation and removal of arsenic, accomplished simultaneously with the biological iron oxidation. In addition, other more conventional methods for arsenic removal are also discussed, such as iron or alum coagulation, lime softening, ion exchange, activated alumina and membrane separation processes. The aforementioned methods have been comparatively evaluated and the relevant conclusions have been drawn with respect to the applicability of arsenic treatment methods, depending on certain parameters, such as locally varying water quality characteristics, sustainability, and economic feasibility. PMID- 16700427 TI - Polychlorinated biphenyls (PCBs): routes of exposure and effects on human health. AB - The polychlorinated biphenyls (PCBs) are synthetic organochlorine chemicals that were useful industrial products in the past, but their production was ended because they persist in both the environment and living organisms. The PCBs are mixtures of up to 209 different components (congeners), depending on the number and position of chlorines around the biphenyl ring. The PCBs are fat-soluble substances to which everyone is exposed through ingesting animal fats, inhalation, or dermal contact. Exposure to PCBs suppresses the immune system, thereby increasing the risk of acquiring several human diseases. Both ortho substituted and coplanar (dioxin-like) congeners are tumor promoters that enhance the effects of other carcinogenic substances. PCB exposure, especially during fetal and early life, reduces IQ and alters behavior. The PCBs alter thyroid and reproductive function in both males and females and increase the risk of developing cardiovascular and liver disease and diabetes. Women are at high risk of giving birth to infants of low birth weight, who are at high lifetime risk for several diseases. As knowledge of their toxic effects has grown faster than environmental levels have declined, PCBs remain dangerous contaminants. PMID- 16700429 TI - Phytoextraction and phytofiltration of arsenic. AB - Arsenic, a ubiquitous contaminant in groundwater and soils, is currently drawing much public attention. Arsenic-contaminated soils can be cleaned up via phytoextraction-the use of plants to extract the arsenic from soil and transport it into aboveground tissues. Arsenic removal from polluted soils can be carried out using hyperaccumulator ferns like the Chinese brake fern Pteris vittata, which accumulates very high concentrations of the element in aboveground tissues. The capacity of the plant to take up large concentrations of arsenic, even at low levels in soil, illustrates efficient bioaccumulation. The possibility of using Pteris ferns to remove arsenic from water by phytofiltration has been proposed. PMID- 16700430 TI - Methyl parathion: an organophosphate insecticide not quite forgotten. AB - Methyl parathion (MP), a toxic organophosphate insecticide approved for outdoor use only, is classified by the World Health Organization (WHO) as a Category Ia (extremely toxic) and by the United States Environmental Protection Agency (U.S. EPA) as a Toxicity Category I (most toxic) insecticide. In several U.S. states in the late 1980s and early 1990s, toxic exposures were created by the illegal use of MP indoors by uncertified pest control operators. As the health effects of MP exposure became evident with increasing public awareness, intervention by the U.S. government, in collaboration with several agencies and public initiatives, led to investigations of MP exposure. After evidence of MP metabolites from urine samples confirmed the exposure, in 1998 the indoor use of MP was banned in the U.S. to protect human health, especially that of children, and the environment. Toxic exposures to MP also occurred in developing countries. In El Salvador, occupational exposure to MP in farmers introduced environmental exposures among agricultural families, who presented with the cholinergic features of MP toxicity. Suicidal MP poisoning was reported in Nepal. A fatal accidental poisoning in children in Peru reflected the serious health risk of pesticides in developing countries. The negligence of pesticide exporters raised human rights issues over the tragedy. Nevertheless, MP exposure remains a potential health risk in both the U.S. and the developing world. Preventive measures in reducing the use of toxic chemicals should be taken seriously to protect human health and the environment. PMID- 16700431 TI - Gender and ethnic differences in urban and rural first-contact schizophrenia outpatients in Trinidad. AB - This study investigated gender and ethnic differences in the rate of first contact outpatients with schizophrenia in the setting of a more-urban region (MUR) and a less-urban region (LUR) in Trinidad. In a prospective study, 134 first-contact patients with a diagnosis of schizophrenia were selected from two ecologically different regions. RESULTS: Of this population, 56.7% were of African origin and 32.1% were of Indian descent. Gender differences were significant, with males accounting for 66.4% (n=89) of patients with schizophrenia (chi2 = 14.45, d.f. = 1, p = 0.0001). Further analysis by age categories revealed a significant male predominance at ages 20-24 (p = 0.0001) and 25-29 (p = 0.002). Young African males (15-19 y, p = 0.049) predominated in MUR compared with LUR. The results showed a marked presence of Afro-Trinidadian males in both outpatient clinics (p < 0.05). We conclude that gender and ethnicity are important variables in the presentation of schizophrenia in Trinidad, whereas neither rural nor urban environments appeared to influence the expression of schizophrenia. PMID- 16700432 TI - Lies. PMID- 16700433 TI - Positional plagiocephaly: pathogenesis, diagnosis, and management. AB - Positional plagiocephaly is a deformation resulting from intrauterine constraint or postnatal positioning leading to asymmetrical cranial growth. There has been a steady increase in referrals for positional plagiocephaly following the release of the American Academy of Pediatrics recommendation of supine infant sleeping position to prevent Sudden Infant Death Syndrome (SIDS) in 1992, largely because of poor parent education on the risks of prolonged occipital pressures. While this deformity is fairly easy to manage when diagnosed early, treatment can become more difficult and complicated with prolonged course. Because of this, it is essential that primary care physicians and parents be educated on recognition of positional plagiocephaly, prevention strategies, and treatment options. In milder cases, where diagnosis is made early, the deformation can be managed by stretching exercises and regular prone positioning, while in more severe cases molding helmets may be needed. Following appropriate treatment, success rates for acceptable cranial shape may be as high as 92%. PMID- 16700434 TI - Increasing awareness of obesity as a treatable medical condition. AB - OBJECTIVE: To estimate the prevalence of overweight and obesity in a population comprised of patients seen at two local primary care clinics and to evaluate the management of this epidemic in a primary care setting. METHODS: Patient medical charts from two urban community clinics in Louisville, Kentucky, were reviewed to determine how the diagnosis of overweight and obesity was assessed and treated. RESULTS: The estimated prevalence of obesity in this population was 40.2% as compared to the national statistics of 30.5%. Despite the higher percentage of obesity, less advice regarding weight loss was given to the patient group under study (30.5%) compared to national survey estimates (42% advised to lose weight and 34% counseled on physical activity). CONCLUSIONS: Data presented here suggest that patients within this Kentucky urban primary care community have not received adequate obesity advice from their physician. Obesity is a growing concern and must be addressed by increasing awareness. PMID- 16700435 TI - The Kentucky medical curriculum: continuing innovations in educating physicians. AB - The University of Kentucky College of Medicine (UKCOM) retains a long history of educational commitment, quality, and innovation. Since undergoing a major curricular revision in the early 1990s, the evolving UKCOM curriculum has continued to incorporate advances in biomedical knowledge and pedagogy while meeting changing societal needs and expectations for physicians in practice. Building upon its established record of excellence in medical education, a curricular quality assurance (QA) program has been initiated to more efficiently guide improvement and innovation by providing faculty with key resources to identify and disseminate local best practices in teaching, learning, and evaluation. Through such efforts, the University of Kentucky College of Medicine is committed to providing a rich educational experience that will meet the needs of students, physicians, and patients well into the 21st century. PMID- 16700436 TI - Where is the plan? PMID- 16700437 TI - Bleeding Kentucky: unjust tort laws cause a hemorrhage of physicians to other states. PMID- 16700440 TI - Improving health care quality: national policy and local practice. PMID- 16700438 TI - The role of psychiatrists and the Oregon Death with Dignity Act. PMID- 16700442 TI - Are we preparing health services administration students to respond to bioterrorism and mass casualty management? AB - Bioterrorism/natural disaster events add significant specialized demands and disrupt normal operation of the health system, often for an indefinite period of time. Health administration leaders should be educationally prepared for and informed about these potential events, but few receive this knowledge via their academic preparation in health administration. This study examined the existence of coverage of bioterrorism topics in health administration curricula and characteristics of bioterrorism coverage in current health administration programs through a self-completed survey among AUPHA graduate and undergraduate program members. Of the total survey respondents, only 32% of programs have current coverage of bioterrorism. The main reasons for nothavingbioterrorism coverage were not having enough resources; not having enough time to develop course/materials; and not thinking it is necessary to add these courses/materials. To prepare better and to inform future health administrators regarding major disruptive circumstances, advocacy and documentation are important to develop and implement bioterrorism awareness. Possibly, suggested minimum curricular requirements, content, and mechanisms for inclusion can be developed in the near future. Health administration educators should address the new reality and demonstrate that their graduates can function and lead in crises and situations disruptive to normal commerce. PMID- 16700441 TI - Competency-based healthcare management education: the Saint Louis University experience. AB - As a follow up to a school-wide initiative to create a common set of competencies for all degree programs in the Saint Louis University School of Public Health, in January 2000 the Department of Health Management and Policy (HMP, renamed from the Department of Health Administration in 2002) began a process to develop a competency-based curriculum for its Master of Health Administration (MHA) degree program with the goal of establishing a foundation for systematically measuring the learning outcomes of its students as they progressed through the program. This article describes how the department developed a set of competencies most appropriate for graduate training in healthcare management, how it incorporated these into its overall MHA program curriculum and content, and how effective this approach has been in measuring student progress in mastering these competencies over the first two years of this initiative. The problems and challenges encountered during this process are discussed, as are the next steps for effectively using competencies to assess healthcare management program learning outcomes. Our experience provides a model for other healthcare management programs considering using an outcomes approach for curriculum development and assessment. PMID- 16700443 TI - E-service learning: A pedagogic innovation for healthcare management education. AB - This paper proposes an innovation in service learning that we identify as e service learning. By adding the "e" to service learning, we create a service learning model that is dynamic, mediated by technology, and delivered online. This paper begins by examining service learning, which is a distinct learning concept. Service learning furnishes students with opportunities for applied learning through participation in projects and activities in community organizations. The authors then define and conceptualize e-service learning, including the anticipated outcomes of implementation such as enhanced access, quality, and cost effectiveness of healthcare management education. Because e service learning is mediated by technology, we identify state of the art technologies that support e-service learning activities. In addition, possible e service learning projects and activities that may be included in healthcare management courses such as finance, human resources, quality, service management/marketing and strategy are identified. Finally, opportunities for future research are suggested. PMID- 16700444 TI - Executive competencies in healthcare administration: preceptors of the Army Baylor University Graduate Program. AB - The purpose of this research was to identify the mentoring and executive competencies required among preceptors of the Army-Baylor University Graduate Program in Health and Business Administration, and to specify the requisite skills, knowledge, and abilities (SKAs) needed to achieve those competencies. In the first wave of inquiry, a list of 123 competencies and associated SKAs was elicited from a network of 80 current and past preceptor executives employing a Delphi methodology using e-mail. An expert panel, which consisted of seven past program directors, examined and sorted the list into four preceptor content domains, viz., Health Systems Management (HS Management), Leadership, Residency Administration, and Community Involvement. Frequency analyses showed that the HS Management domain constituted over half of the competencies, with particular emphasis on strategic thinking, planning, billing, finance, manpower, and contracting. In the second wave, the preceptor Delphi network reviewed the expertpanel list and made 7-pointSKA importance ratings on an 80-item structured questionnaire representative of the four domains. Findings indicated thataverage SKA ratings were reliable and agreed upon to a high degree among preceptors. Results, rank ordered by SKA item means within preceptor content domains and overall, suggested that the most important rated items centered on teamwork, negotiation, interpersonal skills, communication, leadership vision, and customer and healthcare business operations. Outcomes from the competency list are expected to be useful for preceptor mentoring, self-assessment, and for professional development. Additionally, specific SKAs can provide a means for developing job requirements and career performance criteria at a behavioral task level, and can contribute information for identifying continuing education and conference topical needs. PMID- 16700445 TI - A powerful ally. PMID- 16700447 TI - Long-term care partnerships can relieve Medicaid's growing burden. PMID- 16700446 TI - Taking the pulse of the State of the Union. PMID- 16700448 TI - Implementing personal health records. PMID- 16700449 TI - Claims data as a viable treatment resource. PMID- 16700450 TI - Reaching out on obesity. PMID- 16700451 TI - A medical approach to the diagnosis and treatment of dental caries. PMID- 16700452 TI - Concierge care can cut costs. PMID- 16700453 TI - Avoiding misrepresentation of medical history. PMID- 16700454 TI - Patient self-management tools. PMID- 16700455 TI - Some surprises in public perception project research. PMID- 16700456 TI - Meeting community needs: a hallmark of Catholic health care yesterday, today, and tomorrow. PMID- 16700457 TI - The ministry unites to better "tell the good news" of community benefits. PMID- 16700458 TI - One doctor's spiritual Journey (so far). AB - In 2003, the author of this article, who is on the staff of a Tennessee hospital, entered St. Louis's Aquinas Institute of Theology's program in health care mission, seeking to fulfill his longing for greater personal spirituality. Three years later, he is preparing to graduate with a master's degree, and is looking forward to sharing what he has learned with fellow physicians and others. The author credits the program with offering him a much broader understanding of why Catholic health care is a ministry, not just a not-for-profit enterprise. He sees his work at St. Mary's as an exercise in spirituality, and he sees his profession as a personal ministry. He now wants to share what he has learned. The author, who has helped initiate new programs for physician spirituality at his hospital, also speaks at community events about the healing mission of Catholic health care. He hopes that all doctors will reclaim their ancient tradition of providing compassionate care for poor, underserved, and vulnerable persons, and seek to be healing presences to those they serve. PMID- 16700459 TI - Finding peace and joy in the practice of medicine. AB - Although they are healers, physicians are not themselves immune from "secondary stress." In fact, they may be more susceptible to it because they tend not to acknowledge the fact that it exists. In this article, the author, a hospice physician who is a former nurse, explains her personal strategy for combating secondary stress: increased emphasis on maintaining a healthy personal spirituality. Her approach includes morning reflection during her morning fitness activities, making time for family and leisure activities, and, most importantly, incorporating spirituality into her daily work. Bringing her spiritual self to the office and invoking spirituality at the "right" time--not just at set, convenient times--has helped her deal with today's challenges: time pressures, the threat of litigation, reimbursement issues, and misaligned incentives. PMID- 16700460 TI - Making room for spirituality. AB - Avera Health, Sioux Falls, SD, is committed to providing opportunities for physicians to grow spiritually as well as professionally. This commitment has resulted in several initiatives, among which are the formation of several spirituality/reflection groups for doctors and the practice of beginning physician council meetings with sessions on mission. In a round-robin discussion accompanying the article, several leaders of physician spirituality/reflection groups explain how Avera's emphasis on spiritually has helped them avoid burnout and become better doctors. A list of suggested readings used by the reflection groups reveals the diversity of topics that can fall under the heading of "spirituality". PMID- 16700461 TI - Creating healing environments. AB - In 2004 two organizations, the George Washington Institute for Spirituality and Health, Washington, DC, and the Supportive Care Coalition: Pursuing Excellence in Palliative Care, Portland, OR, collaborated in an experiment seeking antidotes to the depersonalization of health care. Their "Hospital-Based Spirituality Initiative: Creating Healing Environments" was intended to achieve two objectives: First, to develop and test strategies that encourage clinical caregivers to attend to patients' spiritual concerns; and, second, to better understand the organizational values and infrastructure that support increasing the spiritual care that caregivers provide. The initiative was conducted in five faith-based hospitals and two secular hospitals. Evaluation of the data indicated that the initiative was success for patients and caregivers. PMID- 16700462 TI - Founding physicians. AB - Physicians have always been key contributors to the founding and success of Catholic health care facilities in the United States. Their contributions have ranged from working with clergy to attract a congregation of women religious to open a facility to raising funds for hospital buildings or donating medical equipment. For a century and a half, doctors have partnered with women and men religious to improve the health and quality of communities. Pioneers in their own right, these physicians should be considered "physician founders" of the Catholic health ministry. PMID- 16700463 TI - Canon law and ethical and religious directives. PMID- 16700464 TI - Justice and genetics: whose Holy Grail? PMID- 16700465 TI - Genomics and the ministry: the executive perspective. Four leaders of Catholic health care organizations offer their opinions. PMID- 16700466 TI - "Co-creating" the space for change. PMID- 16700467 TI - Nurturing the institution's soul. PMID- 16700468 TI - Managing anthelmintic resistance in small ruminant livestock of resource-poor farmers in South Africa. AB - Gastrointestinal parasitism is one of the most important disease complexes of sheep and goats impacting on the resource-poor livestock farmer. Of the responsible nematodes, Haemonchus contortus, a blood-sucking worm of the abomasum, poses possibly the greatest threat. Over the past several decades, the worm has been controlled through the use of anthelmintics, but the emergence of anthelmintic resistance has threatened this chemotherapeutic approach. In Africa, the overall prevalence of anthelmintic resistance has not been extensively investigated, particularly within the resource-poor farming sector, but resistance has been reported from at least 14 countries with most of the reports emanating from Kenya and South Africa and the majority concerning H. contortus. While levels of resistance under commercial sheep farming systems in South Africa is considered to be amongst the worst in the world, resistance has also been reported from the resource-poor farming sector. Increases in productivity and reproduction of livestock and the development of markets for sale of animals are seen by international funding bodies as a way out of poverty for communities that keep livestock. This must lead to the greater need for parasite control. At such times, the risk of levels of anthelmintic resistance escalating is much greater and there is therefore a need to look at alternatives to their use. Proposed strategies include the appropriate, but judicious use of anthelmintics by application of the FAMACHA system and the use of alternatives to anthelmintics such as strategic nutrient supplementation. It is also very clear that there is a strong demand for knowledge about animal diseases, including helminthosis, and their effective management in the resource-poor livestock farming communities. This is an important challenge to meet. PMID- 16700469 TI - The presence of quill mites (Gabucinia bicaudata) and lice (Struthiolipeurus struthionis) in ostrich wing feathers. AB - Quill mites (Gabucinia bicaudata) and lice (Struthiolipeurus struthionis) may infest ostrich feathers, resulting in skin damage, pruritus and excessive feather preening and loss. Four different feather types (prime white, femina extra wide, femina class 1, and femina short; n = 10) were collected. The quill mites and lice were removed with fine forceps, studied using a photographic optical microscope and counted microscopically at x 100 magnification following collection by sedimentation. They were placed in separate Petri dishes containing lactophenol solution and examined (x40 magnification). Anatomical features are described. The density of quill mites in all feather types of both wings was higher than that of the lice. There was no significant difference between the counts of both arthropods on the left wing and the right wing, respectively, except for the femina class 1 quill mites (P = 0.01). The femina extra wide feathers were a preferred habitat in both wings. Large standard deviations (quill mites left wing: 73 +/- 8; quill mites right wing: 69 +/- 7) suggested variations in the degree of migration between feather shafts or as a response to escape preening. It is recommended that ostriches be treated with an oral preparation of Ivermectin administered per os at a dosage rate of 0.2 mg/kg at 30-day intervals for quill mites, and with a 1-5 % Malathion dust at 14-day intervals for lice. PMID- 16700470 TI - Xylazine, diazepam and midazolam premedicated ketamine anaesthesia in white Leghorn cockerels for typhlectomy. AB - Thee different combinations of ketamine hydrochloride were used to induce general anaesthesia for surgical operations (typhlectomy) in 30 adult, single-comb White Leghorn cockerels. They were randomly divided into three groups, each comprising 10 birds. Birds in Group I received xylazine-ketamine combinations at the dose rate of 2 mg xylazine and 10 mg ketamine per kg i.v., whereas birds of Group II received diazepam (2.5 mg/kg i.v.) and 5 min later ketamine (75 mg/kg i.m.). In the Group III, midazolam (2 mg/kg i.m.) and 5 min later ketamine (50 mg/kg i.v.) was administered. The onset of sedation/anaesthesia was shortest (1.60 +/- 0.27 min) in Group I, followed by Group II (8.40 +/- 0.83 min) and Group III (17.10 +/ 1.71 min). Recovery period was shortest in the Group I (65-75 min) followed by Group II (80-85 min) and Group III (92-105 min). Sedation, muscle relaxation and surgical anaesthesia was optimal and excellent in Group I compared with the other two groups. Torticollis, salivation and dyspnoea were observed in Group III. Short-term limb contractions were present in all birds in Groups II and III, up to 20 min of observation. Recovery from anaesthesia was smooth in all three groups. A Surgical procedure (typhlectomy) was performed on all birds. Hypothermia was observed in Group II, whereas heart and respiratory depression was recorded in Group I. Blood sugar level did not vary significantly in any anaesthetic regime. The reduction of haemoglobin was maximum in Group II compared with Groups I and III. Hypoxaemia and hypercapnaea were elevated in all birds in Groups II and III. Blood electrolytes did not vary significantly from the baseline values among the three groups of birds during the period of observation (120 min). The xylazineketamine combination was found to be the best anaesthesia for surgical intervention in chickens. PMID- 16700471 TI - Bone mineral response to ammonium sulphate offered as a lick supplement in beef calves. AB - Sixteen Bonsmara calves (4 males, 12 females) between 10 and 18 months of age were blocked according to age and sex and randomly assigned to 2 groups. They were offered licks containing bone meal and salt (50:50 ratio) (control) and bone meal and ammonium sulphate (NH4SO4) at 1.25, 2.5, 5, 10, 15, and 18% (treatment) to evaluate the effects of dietary anions on bone phosphate (P) concentration. Bone P concentration was significantly (P < 0.05) higher in the NH4SO4 group compared with the control group, indicating that NH4SO4 was able to increase the P content of bone at each of the 6 concentrations used in the lick relative to the control animals, thereby improving the P status of the animals. Ammonium sulphate at 15% and 18 % in the lick also significantly (P < 0.05) increased bone P compared with the lower concentrations of NH4SO4. Bone calcium (Ca) fluctuated as a result of the acidogenic lick. There was absorption of Ca when P was being resorbed and resorption of Ca when P was being absorbed into and out of bone. Bone Ca:P ratio ranged from 3.2 to 6.4 among the control group and 1.6 to 4.3 among the treatment group. Animals receiving the acidogenic lick had a higher percentage ash compared to the control group for most of the experimental period. Bone magnesium (Mg) fluctuated in response to the acidogenic lick, and it was difficult to show a relationship between bone Mg and Ca or P. The overall mean cortical bone thickness was significantly (P < 0.05) greater in treatment (1.60 mm) compared with control (1.43 mm) calves and this was also true at sampling periods 2, 4, 5 and 6. Bone thickness followed bone P and not bone Ca. Results from this research indicate that the addition of ammonium sulphate to a lick had a beneficial effect in improving the P status by increasing bone P and improving the mineral status of bone by increasing the thickness of cortical bone and percentage ash. PMID- 16700472 TI - Ovine placental eluate immunoglobulins recognise isologous and third party acid treated trophoblast microvesicle antigens in vitro. AB - Placental microvesicles were prepared from ovine placentae and immunoglobulins eluted with 0.5 M glycine buffer pH 2.5. The ability of eluate immunoglobulins to re-associate with isologous (self) and third party acidified microvesicles was tested by ELISA. Ovine placental immunoglobulins re-associated with isologous and third party acidified microvesicles suggesting that at least 2 types of antigenic epitopes I and II maybe expressed on the ovine placentae. Type I antigens may be present on placentae of all ovines while type II epitopes may be paternally derived, hence unique to each pregnancy. Analysis by SDS PAGE revealed the heavy and light chains of IgG at 57 and 27 kDa, respectively, together giving a relative molecular weight of 158 kDa. Results suggest that immunoglobulins produced to placental microvesicle antigens may be directed to some but not all antigenic epitopes expressed on the trophoblast, possibly defining a mechanism by which the foetus evades maternal immunological rejection. PMID- 16700473 TI - Common conditions leading to cattle carcass and offal condemnations at 3 abattoirs in the Western Province of Zambia and their zoonotic implications to consumers. AB - From a total of 32 717 cattle slaughtered, 183 whole carcass condemnations were attributable to 9 diseases and conditions, namely, tuberculosis (TB), cysticercosis, emaciation, generalised lymphadenitis, jaundice, abscesses, moribund, sarcosporidiosis and odour. Bovine TB was the most important cause of condemnations (152/183, 83.1%). Bovine cysticercosis and sarcosporidiosis accounted for 5/183 (2.7%) and 8/183 (4.4%), respectively, while each of the remaining conditions contributed less. Among the many conditions responsible for offal/organ condemnations were fascioliasis, contagious bovine pleuropneumonia, hydatidosis and TB. In terms of number and weight, Fasciola gigantica infections made livers and lungs the most condemned offals (20.1% and 0.7%, respectively). Hydatidosis was the cause of 0.9% lung and 0.1% liver losses. Cysticercus bovis contributed to only 0.05% of all inspected tongues, hearts, and heads. TB was very rare in heads (0.01%). The financial impact of whole carcasses and offals condemned during the study period was enormous and deprived livestock farmers of the much needed revenue and consumers of protein sources. Much or all of the condemned material that could have been useful was wasted by not being retrieved for conversion to processed meat, bone meal or pet food. Failure to detect lesions of potential zoonotic diseases at slaughter poses a health risk to consumers especially when meat is eaten undercooked. PMID- 16700474 TI - A comparative study of the effect of 2 hormonal treatment protocols on the reproductive performance of previously anoestrous dairy cows. AB - The objective of this study was to compare the reproductive performance of anoestrous dairy cows treated just prior to the mating start date (MSD) with a Controlled Internal Drug Releasing Device (CIDR) which was placed intravaginaly for either 6 or 8 days, and a combination of oestradiol benzoate. Lactating dairy cows (n = 926) that had been diagnosed with anovulatory anoestrus were divided into two sub-groups. A hormonal treatment protocol that involved the use of a CIDR device containing 1.9 g of progesterone was inserted into the vagina of each cow and left intravaginally for either 6 (6-day group, n = 441) or 8 days (8-day group, n = 485). Every cow in the trial was inseminated after being detected in oestrus from Day -2 onwards (where Day 0, was a herd's MSD), using an appropriate detection aid according to the herds' preference. Cows that had been seen in oestrus and were inseminated by Day 2 were selected for re-synchrony. The standardised re-synchrony involved the re-insertion of a previously used CIDR device into the vagina of each cow on Day 14, together with an injection of 1.0 mg oestradiolbenzoate i.m. This CIDR device was removed on Day 22 and each of these cows injected with 1.0 mg oestradiol benzoate i.m. on Day 23. Each re synchronised cow that was detected in oestrus was re-inseminated. Treatment with an 8-day CIDR increased the proportion of cows submitted for insemination within the first 3 days of the MSD, compared with the 6-day group (83.7 % vs 71.2%, respectively, P < 0.001), as well as the proportion of cows conceiving within the first 3 days of MSD (36.2% vs 27.7%, P = 0.02), but reduced both the interval from MSD to the first service (4.5 +/- 0.5 vs 6.8 +/- 0.7, P = 0.01), and the interval from MSD to conception (28.1 +/- 1.5 vs 34.0 +/- 1.8, P = 0.009). A greater percentage of the cows in the 6-day group that were not pregnant to the first insemination were submitted for a second insemination by Day 28 compared with the 8-day group (81.1% vs 68.3%, P < 0.001). Conception rates for cows submitted for this second insemination by Day 28 of the MSD were also higher in the 6-day than in the 8-day group (48.4% vs 33.9%, P = 0.009). The percentage of cows pregnant at the end of a herd's AI period of 6 weeks did not differ (57.1% vs 54.8% for 8-day and 6-day groups, respectively, P = 0.42); neither did the proportion of cows pregnant at the end of the a herd's combined AI and natural mating period of 21 weeks (81.4% vs 79.2%, for 8-day and 6-day groups respectively P = 0.36). Treatment of anovulatory anoestrous dairy cows with a combination of an 8-day CIDR and oestradiol benzoate before the MSD improved their reproductive performance by increasing the portion of cows submitted for insemination within the first 3-days of the MSD by reducing the interval from MSD to first service and by increasing the conception rate to the first insemination to collectively reduce the average interval from MSD to conception. PMID- 16700475 TI - Nephro-cutaneous fistula in a dog. AB - Nephro-cutaneous fistula, although reported in humans, has not been reported in the dog. In humans the majority of cases develop in patients with a history of previous renal surgery, renal trauma, renal tumours, or chronic urinary tract infection with abscess formation. The dog in this report developed a nephro cutaneous fistula secondary to a traumatic induced renal abscess with formation of a draining sinus tract to the exterior of the body. The animal underwent simple nephrectomy, which resulted in complete resolution of the fistula. PMID- 16700476 TI - Partial facial duplication (diprosopus) in a goat kid. AB - The anatomical and clinical features of a live-born diprosopic goat kid are described. The kid had two faces with two eyes each, two complete oral cavities and nostrils and two ears. Caudal to the neck, the kid grossly appeared normal. Both mouths of the kid showed synchronous suckling motions. Elevated respiratory and heart rates were recorded and the temperature was subnormal. Radiological examination showed a single trunk and vertebral column, normal limbs, two sets of jaws, three orbits, and contrast radiography revealed a single patent oesophagus. There was maxillary and mandibular duplication resulting in two faces. There was a cleft palate. The oropharyngeal regions of each face merged to form a single laryngopharynx and oesophagus. There was a single brain with hypoplasia of the cerebellum. The left and right cerebral hemispheres were fused rostrally, and there was duplication of the optic chiasma and the pituitary gland. The olfactory tract was absent and the superficial origins of most of the cranial nerves were not discernible. PMID- 16700477 TI - Pneumomediastinum, pneumothorax and pneumoretroperitoneum following endoscopic retrieval of a tracheal foreign body from a cat. AB - A 6-year-old entire male cat was presented with a 1-week history of severe dyspnoea without coughing. Upon auscultation, an inspiratory and particularly pronounced expiratory wheeze was noted, with severe dyspnoea. The minimum database was normal. Plain thoracic radiographs showed signs of a mural or intraluminal intrathoracic (T1-T4) tracheal narrowing. A dynamic collapsing trachea was ruled out using fluoroscopy. Bronchoscopy was performed and a dark green and brown spiculated foreign object was found just cranial to the carina. Following removal, the cat rapidly developed extensive truncal subcutaneous emphysema and oxygen-responsive dyspnoea and cyanosis. Follow-up radiographs demonstrated unilateral pneumothorax and lung collapse, marked pneumomediastinum and dissection of air through the tracheal wall. A thoracic drain was placed and the pneumothorax resolved rapidly. Follow-up radiographs demonstrated resolution of pneumothorax and development of extensive retroperitoneal air. The cat made an uneventful recovery. The foreign object was the calyx and stem of a flower. This article emphasises the importance of diagnostic imaging in the dyspnoeic patient, both for confirming initial suspicions of respiratory tract disease, and in managing and charting post-therapy resolution or complications. PMID- 16700478 TI - PCP Profile reports: a Medicaid tool for effective practice management. PMID- 16700479 TI - Be careful about what you sign. PMID- 16700480 TI - Student section report. PMID- 16700481 TI - 2005/2006 Arkansas Medical Society Council report. PMID- 16700483 TI - Pulaski County Medical Society 2005 annual report. PMID- 16700482 TI - Arkansas Department of Health and Human Services Division of Health activity report for 2005. PMID- 16700484 TI - Supporting a culture of evidence. PMID- 16700486 TI - Celebrate nursing. Celebrate the excellence. PMID- 16700485 TI - The other side of the bed. PMID- 16700487 TI - The gift of caring. PMID- 16700494 TI - [Maternity and handicap. Modifying our attitude and introducing practical responses]. PMID- 16700495 TI - [Maternity and handicap, a taboo?]. PMID- 16700496 TI - [Maternity and handicap, legislative aspects]. PMID- 16700497 TI - [Caring for visually handicapped pregnant women]. PMID- 16700498 TI - [Representation of the fetus in utero for visually handicapped women]. PMID- 16700499 TI - [Caring for pregnant deaf women]. PMID- 16700500 TI - [The bear and the masked imp, or an anesthesia story told to children]. PMID- 16700502 TI - [Developing self help groups for parents of burned children]. PMID- 16700503 TI - Peptide substrates of dipeptidyl peptidases. PMID- 16700504 TI - Phosphorus-containing inhibitors of proteolytic enzymes. PMID- 16700505 TI - Biochemical properties of recombinant prolyl dipeptidases DPP-IV and DPP8. PMID- 16700506 TI - Prediction of dipeptidyl peptidase (DP) 8 structure by homology modelling. PMID- 16700507 TI - Structure and function in dipeptidyl peptidase IV and related proteins. PMID- 16700508 TI - Expression and function of dipeptidyl peptidase IV and related enzymes in cancer. PMID- 16700509 TI - DP8 and DP9 have extra-enzymatic roles in cell adhesion, migration and apoptosis. PMID- 16700510 TI - In vivo effects of a potent, selective DPPII inhibitor: UAMC00039 is a possible tool for the elucidation of the physiological function of DPPII. PMID- 16700511 TI - Expression of dipeptidyl peptidase IV-like enzymes in human peripheral blood mononuclear cells. PMID- 16700512 TI - Dipeptidyl peptidase 8 has post-proline dipeptidyl aminopeptidase and prolyl endopeptidase activities. PMID- 16700513 TI - Prolyl endopeptidase cleaves the apoptosis rescue peptide humanin and exhibits an unknown post-cysteine cleavage specificity. PMID- 16700514 TI - Distribution of dipeptidyl peptidase IV-like activity enzymes in canine and porcine tissue sections by RT-PCR. PMID- 16700515 TI - Relative contribution of incretins to the glucose lowering effect of DP IV inhibitors in type 2 diabetes mellitus (T2DM). PMID- 16700516 TI - Dipeptidyl peptidase IV: a molecular switch of vascular actions of neuropeptide Y. PMID- 16700517 TI - Dipeptidylpeptidase IV (DPIV) and alanyl-aminopeptidases (AAPs) as a new target complex for treatment of autoimmune and inflammatory diseases-proof of concept in a mouse model of colitis. PMID- 16700518 TI - Dipeptidyl peptidases and inflammatory bowel disease. PMID- 16700519 TI - Possible role of DP IV inhibitors in acne therapy. PMID- 16700520 TI - Dipeptidyl peptidase-IV activity and/or structure homologs (DASH): contributing factors in the pathogenesis of rheumatic diseases? PMID- 16700521 TI - Dipeptidyl peptidase IV (DP IV, CD26) and aminopeptidase N (APN, CD13) as regulators of T cell function and targets of immunotherapy in CNS inflammation. PMID- 16700522 TI - CD26/DP IV in T cell activation and autoimmunity. PMID- 16700523 TI - DPIV/CD26 and FAP in cancer: a tale of contradictions. PMID- 16700524 TI - Type-II transmembrane prolyl dipeptidases and matrix metalloproteinases in membrane vesicles of active endothelial cells. PMID- 16700525 TI - Extra-enzymatic roles of DPIV and FAP in cell adhesion and migration on collagen and fibronectin. PMID- 16700526 TI - Role of neuropeptide Y and dipeptidyl peptidase IV in regulation of Ewing's sarcoma growth. PMID- 16700527 TI - The role of CD26/DPP IV in preservation of early pulmonary graft function. PMID- 16700528 TI - Light and singlet oxygen in plant defense against pathogens: phototoxic phenalenone phytoalexins. AB - Plants defend themselves from pathogen infections or mechanical injury by a number of mechanisms, including the induced biosynthesis of antimicrobial secondary metabolites. These compounds, termed phytoalexins, represent a very economical way to counteract hazard, because the carbon and energy resources are diverted to phytoalexin synthesis only at the early period of attack and only at its site. The occurrence of phenalenone chromophores in phytoalexins of plants originally nonphototoxic suggests that these plants respond to pathogen attacks by biosynthesizing singlet oxygen photosensitizers able to use solar energy for defense. This concept may have implications for the development of novel crop protection strategies. PMID- 16700529 TI - Pentamethylcyclopentadienyl ruthenium(III) vs hexamethylbenzene ruthenium(II) in sulfur-centered reactivity of their thioether-thiolate and allied complexes. AB - The reactivity features of [Cp*Ru(III){eta(3)-tpdt)}] (7) and [(HMB)Ru(II)(eta(3) tpdt)] (10) {Cp* = eta(5)-C(5)Me(5); HMB = eta(6)-C(6)Me(6); tpdt = 3-thiapentane 1,5-dithiolate, S(CH(2)CH(2)S(-))(2)} are presented, together with selected aspects of their (eta(3)-apdt) analogues 8 and 11 {apdt = 3-azapentane-1,5 dithiolate, HN(CH(2)CH(2)S(-))(2)}. This account will highlight the differences observed in their reactions with metal fragments of compounds of Ru and groups 10 and 11 in various coordination environments and with alkylating agents, including alpha,omega-dibromoalkanes. The mechanistic pathway of the alkylation of 7 will be discussed in some detail. PMID- 16700530 TI - Crystals and crystallinity in polymeric materials. AB - A new view of the concepts of crystallinity and crystals in synthetic macromolecules is discussed. In polymeric materials, crystallinity may be present with the concomitant occurrence of large amounts of structural disorder and in the absence of true three-dimensional long-range order. The structures of semicrystalline polymeric materials are discussed in terms of idealized limit models of crystals, where long-range order may be achieved for structural features that are not necessarily coincident with single atoms and are not necessarily point-centered. Typical examples of non-point-centered structural features are the straight lines corresponding to the chain axes of polymer molecules. This view explains the unusual ability of polymers to crystallize even in the presence of a high degree of structural disorder. PMID- 16700531 TI - New features in pentacoordinate phosphorus chemistry. AB - In reactions centered at phosphorus, whether chemical or biochemical, a pentacoordinate transition state species or an intermediate or a stable molecule is very often encountered. In this Account, recent developments in this important area are discussed and compared with the earlier literature. Particular reference, with results from our laboratory, will be made to the apicophilicity, fluxional behavior, bond parameters, and tetra- vs pentacoordination. It is shown that the familiar apicophilicity rules give an oversimplified picture as demonstrated by several exceptions. Extremities of the P-O or P-N bond distances in PX(4)N and PO(4)N systems as revealed by a database search and possible future directions in this area are also discussed. PMID- 16700532 TI - Fabricating and controlling molecular self-organization at solid surfaces: studies by scanning tunneling microscopy. AB - This account presents a summary of recent work describing the control and fabrication of self-organized molecular adlayers on solid substrates. These results demonstrate that molecules, under appropriate conditions, will self organize into well-ordered monolayers on various solid surfaces. Using scanning tunneling microscopy (STM) to probe the structure of these molecular architectures, it is possible to determine the surface quality to single molecule resolution. The surface structures can be controlled by external stimuli such as electrode potential and UV-light. The ability to control how these adlayers form is important for constructing surface molecular architectures with useful properties. PMID- 16700533 TI - What anions do to N-H-containing receptors. AB - Molecules containing polarized N-H fragments behave as H-bond donors toward anions and are widely used as receptors for recognition and sensing purposes in aprotic solvents (CHCl(3), MeCN, and DMSO). We present examples of receptors containing pyrrole and urea subunits, and we discuss the stability of their H bond complexes with a variety of anions. It is demonstrated that the stability of the 1:1 complexes is strictly related to the acidic tendencies of the receptor and to the basic properties of the anion. It may happen also that more basic anions induce the deprotonation of the receptor, if equipped with electron withdrawing substituents. This is typically observed on interaction with fluoride, due to the formation of the very stable [HF(2)](-) self-complex. For urea-based receptors armed with chromogenic substituents, the addition of a large excess of the anion (F(-), OH(-)) may induce the consecutive deprotonation of both N-H fragments, processes signaled by the development of vivid colors. PMID- 16700534 TI - The phomactins. A novel group of terpenoid platelet activating factor antagonists related biogenetically to the taxanes. AB - A description of the structurally unusual "phomactin" family of platelet activating factor antagonists recently found in the marine fungus Phoma sp. is presented. The phomactins show an interesting structural and biosynthetic relationship with the more familiar taxane group of antitumor compounds isolated from yew trees. The Account highlights and discusses this unique relationship and also presents a cogent picture of plausible biogenetic interrelationships within the family of phomactins. Complementary synthetic endeavors with the phomactins are interwoven in the discussions, alongside contemporaneous biosynthetic studies with both the phomactins and the taxanes. PMID- 16700536 TI - Trapping of the S2 to S3 state intermediate of the oxygen-evolving complex of photosystem II. AB - Photosystem II preparations poised in the S(2)...Q(A) state produce no detectable intermediate during straightforward illumination at liquid helium temperatures. However, upon flash illumination in the range of 77-190 K, they produce a transient state which at -10 degrees C advances to S(3) or after rapid cooling to 10 K gives rise to a 116 G wide metalloradical EPR signal. The latter decays with half-times on the order of a few minutes, presumably by charge recombination, and can be regenerated repeatedly by illumination at 10 K. The constraints for Tyr Z oxidation are attributed to the presence of excess positive charge in S(2). Elevated temperatures are required presumably to overcome a thermal barrier in the deprotonation of Tyr Z(+) or most likely to allow secondary proton transfer away from the base partner of Tyr Z. Treatment with 5% (v/v) MeOH appears to remove the constraints for Tyr Z oxidation, and a 160 G wide metalloradical EPR signal is produced by illumination at 10 K, which decays with a half-time of ca. 80 s. Formation of the metalloradical signals is accompanied by reversible changes in the Mn multiline signal. The intermediates are assigned to Tyr Z(*) magnetically interacting with the Mn cluster in S(2), S(2)Y(Z)(*). A molecular model which extends an earlier suggestion and provides a plausible explanation of a number of observations, including the binding of small molecules to the Mn cluster, is presented. PMID- 16700537 TI - Measurement of multisite oxidation kinetics reveals an active site conformational change in Spo0F as a result of protein oxidation. AB - When most proteins undergo oxidative damage, they yield a variety of products containing oxidative damage at a large number of sites, most of which are modified substoichiometrically. The resulting complex mixture of products is not amenable to high-resolution structural analyses. The previous methods of structural analysis have relied upon either very generalized structural analyses such as circular dichroism or the creation of a battery of mutants to try to isolate single-residue damage effects. We present a methodology using mass spectrometry to measure the kinetics of oxidation at many sites simultaneously. Previous studies have shown that these kinetics are determined by the chemical nature of the damage site and by the accessibility of that site to the radical. By measuring deviations in the rate of oxidation from the expected pseudo-zero order kinetics, we can detect and characterize local structural changes due to the oxidative damage. We demonstrate the application of this new technique to the Spo0F protein, a regulator of sporulation in Bacillus subtilis. Circular dichroism studies suggest a partial loss of helical structure of Spo0F as a result of oxidative damage. We report that oxidation causes a three-stage conformational change in Spo0F. Furthermore, we find the dramatic structural changes affect only the region surrounding the active site, while the remainder of the structure remains relatively unperturbed. Finally, we are able to determine that the specific oxidation event that triggers the conformational change at the active site of Spo0F occurs at Met81, a partially conserved methionine in the CheY superfamily. PMID- 16700535 TI - Role of receptor tyrosine kinase transmembrane domains in cell signaling and human pathologies. AB - Receptor tyrosine kinases (RTKs) conduct biochemical signals via lateral dimerization in the plasma membrane, and their transmembrane (TM) domains play an important role in the dimerization process. Here we present two models of RTK mediated signaling, and we discuss the role of the TM domains within the framework of these two models. We summarize findings of single-amino acid mutations in RTK TM domains that induce unregulated signaling and, as a consequence, pathological phenotypes. We review the current knowledge of pathology induction mechanisms due to these mutations, focusing on the structural and thermodynamic basis of pathogenic dimer stabilization. PMID- 16700538 TI - Concatenation of cyan and yellow fluorescent proteins for efficient resonance energy transfer. AB - Highly efficient fluorescence resonance energy transfer between cyan(CFP) and yellow fluorescent proteins (YFP), the cyan- and yellow-emitting variants of the Aequorea green fluorescent protein, respectively, was achieved by tightly concatenating the two proteins. After the C-terminus of CFP and the N-terminus of YFP were truncated by 11 and 5 amino acids, respectively, the proteins were fused through a leucine-glutamate dipeptide. The resulting chimeric protein, which we called Cy11.5, exhibited a simple emission spectrum that peaked at 527 nm when the protein was excited at 436 nm. The time-resolved emission of Cy11.5 was measured using a streak camera. After excitation of Cy11.5 with a 400 nm ultrashort pulse, a fast decay of the CFP emission and a concomitant rise of the YFP emission were observed with a lifetime of 66 ps. By contrast, the emission from CFP alone showed a decay component with a lifetime of 2.9 ns. We concluded that in fully folded Cy11.5 molecules, intramolecular FRET occurred with an efficiency of 98%. Importantly, most Cy11.5 molecules were properly folded, and the protein was highly resistant to all of the tested proteases. In living cells, therefore, Cy11.5 behaved as a single fluorescent protein with a broad excitation spectrum. Moreover, Cy11.5 was used as an optical highlighter after photobleaching of YFP. When HeLa cells expressing Cy11.5 were irradiated at 514.5 nm, a 10-fold increase in the 475 nm fluorescence intensity was observed. These features make Cy11.5 useful as an optical highlighter and a new-colored fluorescent protein for multicolor imaging. PMID- 16700539 TI - Fluorine-19 NMR studies on the acid state of the intestinal fatty acid binding protein. AB - The intestinal fatty acid binding protein (IFABP) is composed of two beta-sheets with a large hydrophobic cavity into which ligands bind. After eight 4-(19)F phenylalanines were incorporated into the protein, the acid state of both apo- and holo-IFABP (at pH 2.8 and 2.3) was characterized by means of (1)H NMR diffusion measurements, circular dichroism, and (19)F NMR. Diffusion measurements show a moderately increased hydrodynamic radius while near- and far-UV CD measurements suggest that the acid state has substantial secondary structure as well as persistent tertiary interactions. At pH 2.8, these tertiary interactions have been further characterized by (19)F NMR and show an NOE cross-peak between residues that are located on different beta-strands. Side chain conformational heterogeneity on the millisecond time scale was captured by phase-sensitive (19)F (19)F NOESY. At pH 2.3, native NMR peaks are mostly gone, but the protein can still bind fatty acid to form the holoprotein. An exchange cross-peak of one phenylalanine in the holoprotein is attributed to increased motional freedom of the fatty acid backbone caused by the slight opening of the binding pocket at pH 2.8. In the acid environment Phe128 and Phe17 show dramatic line broadening and chemical shift changes, reflecting greater degrees of motion around these residues. We propose that there is a separation of specific regions of the protein that gives rise to the larger radius of hydration. Temperature and urea unfolding studies indicate that persistent hydrophobic clusters are nativelike and may account for the ability of ligand to bind and induce nativelike structure, even at pH 2.3. PMID- 16700540 TI - Substrates induce conformational changes in human anion exchanger 1 (hAE1) as observed by fluorescence resonance energy transfer. AB - The one-for-one exchange of Cl(-) and HCO(3)(-) ions is catalyzed by human erythrocyte anion exchanger 1 (hAE1) through a ping-pong mechanism whereby the protein exists in two main conformations, with the single anion-binding site exposed at either the cytoplasmic (inner) side (E(i)) or the extracellular side (E(o)), with interconversion between the two states being possible only after anion binding. Steady-state and time-resolved resonance energy transfer (FRET) techniques were used to determine the distance of the binding site for diTBA (bis (1,3-diethylthiobarbituric acid)trimethine oxonol), a high affinity fluorescent oxonol inhibitor of hAE1, from a benchmark site (probably Lys-430) labeled by external fluorescein maleimide (FM). Using red cell ghost membranes, energy transfer distances were measured in media containing different anions between FM as the donor, covalently attached to one monomer, and diTBA as the acceptor, reversibly bound to the adjacent monomer of a hAE1 dimer. Energy transfer increased significantly in chloride or bicarbonate buffers relative to conditions where no transportable anions were present, that is, in citrate buffer. These differences in transfer efficiencies were interpreted in light of the conformational distributions of hAE1 in various buffers and the possible effects of diTBA itself on the distribution. The analysis indicates that the diTBA binding site comes closer to the FM site by approximately 7 A in chloride buffer as compared to that in citrate (or equivalent changes in diTBA orientation occur) because of the effects of anion binding. This provides the first direct physical evidence for structural changes in hAE1 induced by substrates. PMID- 16700542 TI - ESR signal of the iron-sulfur center F(X) and its function in the homodimeric reaction center of Heliobacterium modesticaldum. AB - Electron transfer in the membranes and the type I reaction center (RC) core protein complex isolated from Heliobacterium modesticaldum was studied by optical and ESR spectroscopy. The RC is a homodimer of PshA proteins. In the isolated membranes, illumination at 14 K led to accumulation of a stable ESR signal of the reduced iron-sulfur center F(B)(-) in the presence of dithiothreitol, and an additional 20 min illumination at 230 K induced the spin-interacting F(A)( )/F(B)(-) signal at 14 K. During illumination at 5 K in the presence of dithionite, we detected a new transient signal with the following values: g(z)= 2.040, g(y)= 1.911, and g(x)= 1.896. The signal decayed rapidly with a 10 ms time constant after the flash excitation at 5 K and was attributed to the F(X)(-)-type center, although the signal shape was more symmetrical than that of F(X)(-) in photosystem I. In the purified RC core protein, laser excitation induced the absorption change of a special pair, P800. The flash-induced P800(+) signal recovered with a fast 2-5 ms time constant below 150 K, suggesting charge recombination with F(X)(-). Partial destruction of the RC core protein complex by a brief exposure to air increased the level of the P800(+)A(0)(-) state that gave a lifetime (t(1/2)) of 100 ns at 77 K. The reactions of F(X) and quinone were discussed on the basis of the three-dimensional structural model of RC that predicts the conserved F(X)-binding site and the quinone-binding site, which is more hydrophilic than that in the photosystem I RC. PMID- 16700541 TI - Fatty-acid-binding protein from the flight muscle of Locusta migratoria: evolutionary variations in fatty acid binding. AB - Intracellular lipid-binding proteins have evolved from a common ancestral gene with the appearance of mitochondrial oxidation, to guarantee, for example, transport of fatty acids through the aqueous cytosol to their site of utilization. The mammalian forms of these lipid carriers are structurally well characterized and have been categorized, on the basis of sequence similarities and several typical ligand-binding features, into four subfamilies. Only a single complex structure of an invertebrate fatty-acid-binding protein (FABP) has been reported to date, which reveals a unique ligand-binding arrangement yet unknown in vertebrate FABPs. In the present study, the structure of a second invertebrate FABP (locust muscle) complexed with a fatty acid has been determined on the basis of intermolecular NOE connectivities between the protein and the uniformly (13)C enriched oleate ligand. The resulting ligand conformation, although resembling the closely related mammalian heart- and adipocyte-type FABPs, is characterized by certain binding features that differ significantly from the typical hairpin turn ligand shapes of the latter forms. This is primarily due to an alanine-to leucine substitution in locust FABPs that produces a steric hindrance for ligand binding. A comparison with an FABP from tobacco hornworm larvae furthermore demonstrates that certain amino acid substitutions that appear to be specific for invertebrates decidedly influence the binding arrangement inside the protein cavity. Hence, as a result of these evolutionary variations, invertebrate FABPs may display a much greater diversity in intracellular lipid binding than observed for the mammalian transport proteins, thus possibly providing new insights for the design of modified lipid carriers. PMID- 16700543 TI - Evidence for iron channeling in the Fet3p-Ftr1p high-affinity iron uptake complex in the yeast plasma membrane. AB - In high-affinity iron uptake in the yeast Saccharomyces cerevisiae, Fe(II) is oxidized to Fe(III) by the multicopper oxidase, Fet3p, and the Fe(III) produced is transported into the cell via the iron permease, Ftr1p. These two proteins are likely part of a heterodimeric or higher order complex in the yeast plasma membrane. We provide kinetic evidence that the Fet3p-produced Fe(III) is trafficked to Ftr1p for permeation by a classic metabolite channeling mechanism. We examine the (59)Fe uptake kinetics for a number of complexes containing mutant forms of both Fet3p and Ftr1p and demonstrate that a residue in one protein interacts with one in the other protein along the iron trafficking pathway as would be expected in a channeling process. We show that, as a result of some of these mutations, iron trafficking becomes sensitive to an added Fe(III) chelator that inhibits uptake in a strictly competitive manner. This inhibition is not strongly dependent on the chelator strength, however, suggesting that Fe(III) dissociation from the iron uptake complex, if it occurs, is kinetically slow relative to iron permeation. Metabolite channeling is a common feature of multifunctional enzymes. We constructed the analogous ferroxidase, permease chimera and demonstrate that it supports iron uptake with a kinetic pattern consistent with a channeling mechanism. By analogy to the Fe(III) trafficking that leads to the mineralization of the ferritin core, we propose that ferric iron channeling is a conserved feature of iron homeostasis in aerobic organisms. PMID- 16700544 TI - Mechanistic significance of the preparatory migration of hydrogen atoms around the FeMo-co active site of nitrogenase. AB - The migration of H atoms over S and Fe atoms in the reaction domain of FeMo-co, the active site of nitrogenase, is described and used to explain mechanistic data on the catalyzed reductions of N(2) and C(2)H(2). After electron transfer to FeMo co, H atoms are generated by fast proton supply to S3B (atom labels from structure 1M1N) and migrate vectorially via several pathways from S3B to locations on the FeMo-co face, specifically Fe6, S2B, Fe2, and S2A (calculated reaction profiles are reported). The E(n)H(n) reduction levels (n = 1-4) in the Thorneley-Lowe kinetic-mechanistic schemes are each potential sequences of substructures with different distributions of H atoms. The positions of H atoms influence the binding of substrates N(2) and C(2)H(2), and the bound substrate subsequently blocks further migration of H atoms past the binding site. This model provides a consistent structural interpretation of (a) the two-site reactivity of C(2)H(2) and the differentiation of the high- and low-affinity sites as due to different preparatory H migration; (b) the differing mutual inhibitions of N(2) and C(2)H(2) in wild-type protein; (c) the modified reactivity of the Azotobacter vinelandii alpha-(Gly)69(Ser) mutant with N(2) and C(2)H(2); and (d) the basis for the stereoselectivity of hydrogenation of C(2)D(2) and its loss in some mutant proteins. Some structures for initially bound N(2) and C(2)H(2), and their hydrogenated intermediates, are presented. The key new concept is that binding sites and binding states for substrates and intermediates are characterized not only by their locations on the FeMo-co face but also by the structural and temporal status of the distribution of H atoms over the FeMo-co reaction domain. PMID- 16700545 TI - Surface plasmon resonance analysis of antifungal azoles binding to CYP3A4 with kinetic resolution of multiple binding orientations. AB - The heme-containing cytochrome P450s (CYPs) are a major enzymatic determinant of drug clearance and drug-drug interactions. The CYP3A4 isoform is inhibited by antifungal imidazoles or triazoles, which form low-spin heme iron complexes via formation of a nitrogen-ferric iron coordinate bond. However, CYP3A4 also slowly oxidizes the antifungal itraconazole (ITZ) at a site that is approximately 25 A from the triazole nitrogens, suggesting that large antifungal azoles can adopt multiple orientations within the CYP3A4 active site. Here, we report a surface plasmon resonance (SPR) analysis with kinetic resolution of two binding modes of ITZ, and the related drug ketoconazole (KTZ). SPR reveals a very slow off-rate for one binding orientation. Multiphasic binding kinetics are observed, and one of the two binding components resolved by curve fitting exhibits "equilibrium overshoot". Preloading of CYP3A4 with the heme ligand imidazole abolishes this component of the antifungal azole binding trajectories, and it eliminates the conspicuously slow off-rate. The fractional populations of CYP3A4 complexes corresponding to different drug orientations can be manipulated by altering the duration of the pulse of drug exposure. UV-vis difference absorbance titrations yield low-spin spectra and K(D) values that are consistent with the high-affinity complex resolved by SPR. These results demonstrate that ITZ and KTZ bind in multiple orientations, including a catalytically productive mode and a slowly dissociating inhibitory mode. Most importantly, they provide the first example of a SPR-based method for the kinetic characterization of binding of a drug to any human CYP, including mechanistic insight not available from other methods. PMID- 16700546 TI - Intraprotein electron transfer in a two-domain construct of neuronal nitric oxide synthase: the output state in nitric oxide formation. AB - Intersubunit intraprotein electron transfer (IET) from flavin mononucleotide (FMN) to heme is essential in nitric oxide (NO) synthesis by NO synthase (NOS). Previous crystal structures and functional studies primarily concerned an enzyme conformation, which serves as the input state for reduction of FMN by electrons from NADPH and flavin adenine dinucleotide (FAD) in the reductase domain. To favor the formation of the output state for the subsequent IET from FMN to heme in the oxygenase domain, a novel truncated two-domain oxyFMN construct of rat neuronal NOS (nNOS), in which only the FMN and heme domains were present, was designed and expressed. The kinetics of IET between the FMN and heme domains in the nNOS oxyFMN construct in the presence and absence of added calmodulin (CaM) were directly determined using laser flash photolysis of CO dissociation in comparative studies on partially reduced oxyFMN and single-domain heme oxygenase constructs. The IET rate constant in the presence of CaM (262 s(-)(1)) was increased approximately 10-fold compared to that in the absence of CaM (22 s( )(1)). The effect of CaM on interdomain interactions was further evidenced by electron paramagnetic resonance (EPR) spectra. This work provides the first direct evidence of the CaM control of electron transfer (ET) between FMN and heme domains through facilitation of the FMN/heme interactions in the output state. Therefore, CaM controls IET between heme and FMN domains by a conformational gated mechanism. This is essential in coupling ET in the reductase domain in NOS with NO synthesis in the oxygenase domain. PMID- 16700547 TI - Structural and functional studies on DHC, the diheme cytochrome c from Rhodobacter sphaeroides, and its interaction with SHP, the sphaeroides heme protein. AB - The diheme cytochrome c (DHC) from Rhodobacter sphaeroides is a soluble protein with a mass of 16 kDa that represents a new class of c-type cytochrome [Vandenberghe, I., et al. (1998) Biochemistry 37, 13075-13081]. The gene encoding DHC is associated with another encoding a cytochrome known as SHP (sphaeroides heme protein). It is believed that DHC is the electron donor for SHP, which is known to bind oxygen. To gain further insight into the properties and role of DHC, we have carried out structure-function studies on the protein and examined its interaction with SHP. The crystal structures of native and recombinant DHC have been determined to resolutions of 1.85 and 2.0 A, respectively. The structures show that DHC folds into two distinct domains each containing one heme. While the N-terminal domain is a class I cytochrome c, the C-terminal domain shows no similarity to any existing structures and thus constitutes a novel cytochrome c structural motif. The shortest, edge-to-edge, distance between the heme groups is 10.2 A, and this distance is bridged by Tyr31, thus ensuring fast internal electron transfer. DHC binds strongly to its proposed physiological partner, SHP (K(d) = 0.26 microM in 10 mM HEPES at pH 7.2 and 25 degrees C). However, at higher salt concentrations, the binding becomes much weaker, indicating the importance of electrostatic interactions. DHC is also very efficient in electron transfer to SHP with a second-order rate constant of 1.8 x 10(7) M(-)(1) s(-)(1) (at pH 7.2, 10 degrees C, and I = 500 mM). The reduction potentials of DHC and SHP are also suitably ordered for a favorable reaction with the hemes of DHC showing potentials of -310 and -240 mV, respectively, and that for SHP being -105 mV. These potentials are unaltered upon complex formation. PMID- 16700548 TI - The crystal structure of NAD(P)H quinone oxidoreductase 1 in complex with its potent inhibitor dicoumarol. AB - NAD(P)H quinone oxidoreductase 1 (NQO1) is a ubiquitous flavoenzyme that catalyzes two-electron reduction of quinones to hydroquinones utilizing NAD(P)H as an electron donor. NQO1 binds and stabilizes several short-lived proteins including the tumor suppressors p53 and p73 and the enzyme ornithine decarboxylase (ODC). Dicoumarol is a widely used potent competitive inhibitor of NQO1 enzymatic activity, which competes with NAD(P)H for binding to NQO1. Dicoumarol also disrupts the binding of NQO1 to p53, p73, and ODC and induces their ubiquitin-independent proteasomal degradation. We report here the crystal structure of human NQO1 in complex with dicoumarol at 2.75 A resolution. We have identified the interactions of dicoumarol with the different residues of NQO1 and the conformational changes imposed upon dicoumarol binding. The most prominent conformational changes that occur in the presence of dicoumarol involve Tyr 128 and Phe 232 that are present on the surface of the NQO1 catalytic pocket. On the basis of the comparison of the NQO1 structure in complex with different NQO1 inhibitors and our previous analysis of NQO1 mutants, we propose that the specific conformation of Tyr 128 and Phe 232 is important for NQO1 interaction with p53 and other client proteins. PMID- 16700549 TI - A trade between similar but nonequivalent intrasubunit and intersubunit contacts in Cro dimer evolution. AB - The homodimeric lambda Cro protein has a "ball-and-socket" interface that includes insertion of an aromatic side chain, Phe 58, from each subunit into a cavity in the hydrophobic core of the other subunit. This overlap between the subunit core and dimer interface hypothetically explains the strong dimerization and weak monomer stability of lambda Cro in comparison to homologues. According to a model developed here and in a previous study [LeFevre, K. R., and Cordes, M. H. (2003) Proc. Natl. Acad. Sci. U.S.A. 100, 2345-2350], the socket cavity evolved in part by replacement of a buried tryptophan in an ancestral stable monomer with a smaller side chain (Ala 33 in lambda Cro). The resulting core defect was in effect repaired by insertion of a different side chain (Phe 58) from a second subunit, generating the ball and socket. Consistent with such an evolutionary trade between intrasubunit and intersubunit interactions, we showed in the previous study that restoration of the ancestral Trp 33 in lambda Cro stabilized the monomer and reduced the extent of dimerization. Here, we report the solution structure of a stable lambda Cro monomer containing the Ala33Trp mutation, which confirms that the restored tryptophan fulfills its ancestral role as a core side chain, filling part of the socket cavity occupied by Phe 58 in the wild-type dimer. The structure also reveals, however, that the cavity is not completely filled by Trp 33, suggesting that its formation could have involved multiple mutations that reduced side chain volume. We offer suggestive evidence of a role of mutations at a second position. PMID- 16700551 TI - Structural and kinetic characterization of Escherichia coli TadA, the wobble specific tRNA deaminase. AB - The essential tRNA-specific adenosine deaminase catalyzes the deamination of adenosine to inosine at the wobble position of tRNAs. This modification allows for a single tRNA species to recognize multiple synonymous codons containing A, C, or U in the last (3'-most) position and ensures that all sense codons are appropriately decoded. We report the first combined structural and kinetic characterization of a wobble-specific deaminase. The structure of the Escherichia coli enzyme clearly defines the dimer interface and the coordination of the catalytically essential zinc ion. The structure also identifies the nucleophilic water and highlights residues near the catalytic zinc likely to be involved in recognition and catalysis of polymeric RNA substrates. A minimal 19 nucleotide RNA stem substrate has permitted the first steady-state kinetic characterization of this enzyme (k(cat) = 13 +/- 1 min(-)(1) and K(M) = 0.83 +/- 0.22 microM). A continuous coupled assay was developed to follow the reaction at high concentrations of polynucleotide substrates (>10 microM). This work begins to define the chemical and structural determinants responsible for catalysis and substrate recognition and lays the foundation for detailed mechanistic analysis of this essential enzyme. PMID- 16700552 TI - The biotin regulatory system: kinetic control of a transcriptional switch. AB - An organism's response to environmental and metabolic cues requires communication between transcription regulatory processes and "other" cellular events. In a number of biological control circuits, the communication is carried out by a single multifunctional protein that participates directly in transcription initiation and in at least one other cellular process. Structural studies suggest that the function of these proteins is dictated by the formation of mutually exclusive protein-protein interactions. However, the rules that govern partner, and thus functional switching, are not known. In the Escherichia coli Biotin Regulatory System, the bifunctional protein, BirA, catalyzes post-translational biotin addition to a biotin-dependent carboxylase and binds sequence-specifically to DNA to repress transcription initiation at the biotin biosynthetic operon. Previous structural and modeling studies suggest that BirA function is determined by formation of alternative homo- and heterodimeric protein-protein interactions. In this work, the BirA functional switch is investigated using DNaseI footprinting and MALDI-TOF mass spectrometry. Results of these measurements indicate that BirA can be selectively targeted toward its enzymatic function simply by increasing the kinetic probability of heterodimerization relative to that of homodimerization. Subsequent shifting to the DNA binding function occurs as the pool of heterodimer partner is depleted and homodimerization dominates. The data support a switching mechanism in which BirA's function is dictated by its probability of encountering a particular protein partner. PMID- 16700553 TI - Peptide-binding motifs for the I-Ad MHC class II molecule: alternate pH-dependent binding behavior. AB - The ability of peptides to form stable complexes with MHC class II molecules expressed in the host determines their ability to recruit CD4 T cells during an immune response. In this study, we sought to define the features of the antigenic peptides that control their kinetic stability with I-A(d) because of the diversity of peptides that this molecule is known to present. Peptide dissociation assays indicated that each pocket of I-A(d) displays exquisite sensitivity to side chain structure, size, and charge. Most surprising were results related to the P1 pocket, which has been difficult to define by conventional competition assays. Our studies revealed a considerable degree of specificity in the P1 pocket but also an unexpected degree of structural flexibility. Amino acids with neutral side chains such as Met and the alternatively negatively charged Glu are both highly favored at P1. Interestingly, these two options at the P1 pocket in I-A(d) display dramatically different pH-dependent interactions with the class II molecule. These findings are discussed in the context of a structural model to explain these data and in light of the immunological implications of pH-dependent behavior of class II peptide complexes in acidic endosomal compartments, where DM-catalyzed loading of class II molecules takes place, and at the neutral pH of the APC cell surface, where class II-peptide complexes promote activation of CD4 T cells. PMID- 16700550 TI - Effects of biological oxidants on the catalytic activity and structure of group VIA phospholipase A2. AB - Group VIA phospholipase A(2) (iPLA(2)beta) is expressed in phagocytes, vascular cells, pancreatic islet beta-cells, neurons, and other cells and plays roles in transcriptional regulation, cell proliferation, apoptosis, secretion, and other events. A bromoenol lactone (BEL) suicide substrate used to study iPLA(2)beta functions inactivates iPLA(2)beta by alkylating Cys thiols. Because thiol redox reactions are important in signaling and some cells that express iPLA(2)beta produce biological oxidants, iPLA(2)beta might be subject to redox regulation. We report that biological concentrations of H(2)O(2), NO, and HOCl inactivate iPLA(2)beta, and this can be partially reversed by dithiothreitol (DTT). Oxidant treated iPLA(2)beta modifications were studied by LC-MS/MS analyses of tryptic digests and included DTT-reversible events, e.g., formation of disulfide bonds and sulfenic acids, and others not so reversed, e.g., formation of sulfonic acids, Trp oxides, and Met sulfoxides. W(460) oxidation could cause irreversible inactivation because it is near the lipase consensus sequence ((463)GTSTG(467)), and site-directed mutagenesis of W(460) yields active mutant enzymes that exhibit no DTT-irreversible oxidative inactivation. Cys651-sulfenic acid formation could be one DTT-reversible inactivation event because Cys651 modification correlates closely with activity loss and its mutagenesis reduces sensitivity to inhibition. Intermolecular disulfide bond formation might also cause reversible inactivation because oxidant-treated iPLA(2)beta contains DTT-reducible oligomers, and oligomerization occurs with time- and temperature-dependent iPLA(2)beta inactivation that is attenuated by DTT or ATP. Subjecting insulinoma cells to oxidative stress induces iPLA(2)beta oligomerization, loss of activity, and subcellular redistribution and reduces the rate of release of arachidonate from phospholipids. These findings raise the possibility that redox reactions affect iPLA(2)beta functions. PMID- 16700554 TI - Characterization of the hydrodynamic properties of the folding transition state of an SH3 domain by magnetization transfer NMR spectroscopy. AB - Protein folding kinetic data have been obtained for the marginally stable N terminal Src homology 3 domain of the Drosophila protein drk (drkN SH3) in an investigation of the hydrodynamic properties of its folding transition state. Due to the presence of NMR resonances of both folded and unfolded states at equilibrium, kinetic data can be derived from NMR magnetization transfer techniques under equilibrium conditions. Kinetic analysis as a function of urea (less than approximately 1 M) and glycerol enables determination of alpha values, measures of the energetic sensitivity of the transition state to the perturbation relative to the end states of the protein folding reaction (the folded and unfolded states). Both end states have previously been studied experimentally by NMR spectroscopic and other biophysical methods in great detail and under nondenaturing conditions. Combining these results with the kinetic folding data obtained here, we can characterize the folding transition state without requiring empirical models for the unfolded state structure. We are thus able to give a reliable measure of the solvent-accessible surface area of the transition state of the drkN SH3 domain (4730 +/- 360 A(2)) based on urea titration data. Glycerol titration data give similar results and additionally demonstrate that folding of this SH3 domain is dependent on solvent viscosity, which is indicative of at least partial hydration of the transition state. Because SH3 domains appear to fold by a common folding mechanism, the data presented here provide valuable insight into the transition states of the drkN and other SH3 domains. PMID- 16700555 TI - The core of tau-paired helical filaments studied by scanning transmission electron microscopy and limited proteolysis. AB - In Alzheimer's disease and frontotemporal dementias the microtubule-associated protein tau forms intracellular paired helical filaments (PHFs). The filaments formed in vivo consist mainly of full-length molecules of the six different isoforms present in adult brain. The substructure of the PHF core is still elusive. Here we applied scanning transmission electron microscopy (STEM) and limited proteolysis to probe the mass distribution of PHFs and their surface exposure. Tau filaments assembled from the three repeat domain have a mass per length (MPL) of approximately 60 kDa/nm and filaments from full-length tau (htau40DeltaK280 mutant) have approximately 160 kDa/nm, compared with approximately 130 kDa/nm for PHFs from Alzheimer's brain. Polyanionic cofactors such as heparin accelerate assembly but are not incorporated into PHFs. Limited proteolysis combined with N-terminal sequencing and mass spectrometry of fragments reveals a protease-sensitive N-terminal half and semiresistant PHF core starting in the first repeat and reaching to the C-terminus of tau. Continued proteolysis leads to a fragment starting at the end of the first repeat and ending in the fourth repeat. PHFs from tau isoforms with four repeats revealed an additional cleavage site within the middle of the second repeat. Probing the PHFs with antibodies detecting epitopes either over longer stretches in the C-terminal half of tau or in the fourth repeat revealed that they grow in a polar manner. These data describe the physical parameters of the PHFs and enabled us to build a model of the molecular arrangement within the filamentous structures. PMID- 16700556 TI - Phi-value analysis of apo-azurin folding: comparison between experiment and theory. AB - Pseudomonas aeruginosa azurin is a 128-residue beta-sandwich metalloprotein; in vitro kinetic experiments have shown that it folds in a two-state reaction. Here, we used a variational free energy functional to calculate the characteristics of the transition state ensemble (TSE) for folding of the apo-form of P. aeruginosa azurin and investigate how it responds to thermal and mutational changes. The variational method directly yields predicted chevron plots for wild-type and mutant apo-forms of azurin. In parallel, we performed in vitro kinetic-folding experiments on the same set of azurin variants using chemical perturbation. Like the wild-type protein, all apo-variants fold in apparent two-state reactions both in calculations and in stopped-flow mixing experiments. Comparisons of phi (phi) values determined from the experimental and theoretical chevron parameters reveal an excellent agreement for most positions, indicating a polarized, highly structured TSE for folding of P. aeruginosa apo-azurin. We also demonstrate that careful analysis of side-chain interactions is necessary for appropriate theoretical description of core mutants. PMID- 16700557 TI - Oxalone and lactone moieties of podophyllotoxin exhibit properties of both the B and C rings of colchicine in its binding with tubulin. AB - Thermodynamics of podophyllotoxin binding to tubulin and its multiple points of attachment with tubulin has been studied in detail using isothermal titration calorimetry. The calorimetric enthalpy of the association of podophyllotoxin with tubulin is negative and occurs with a negative heat capacity change (DeltaC(p) = 2.47 kJ mol(-)(1) K(-)(1)). The binding is unique with a simultaneous participation of both hydrophobic and hydrogen-bonding forces with unfavorable negative entropic contribution at higher temperature, favored with an enthalpy entropy compensation. Interestingly, the binding of 2-methoxy-5-(2',3',4' trimethoxyphenyl)tropone (AC, a colchicine analogue without the B ring) with tubulin is enthalpy-favored. However, the podophyllotoxin-tubulin association depending upon the temperature of the reaction has a favorable entropic and enthalpic component, which resembles both B- and C-ring properties of colchicine. On the basis of the crystal structure of the podophyllotoxin-tubulin complex, distance calculations have indicated a possible interaction between threonine 179 of alpha-tubulin and the hydroxy group on the D ring of podophyllotoxin. To confirm the involvement of the oxalone moiety as well as the lactone ring of podophyllotoxin in tubulin binding, analogues of podophyllotoxin are synthesized with methoxy substitution at the 4' position of ring D along with its isomer and another analogue epimerized at ring E. From these results, involvement of oxalone as well as the lactone ring of the drug in a specific orientation inclusive of ring A is indicated for podophyllotoxin-tubulin binding. Therefore, podophyllotoxin, like colchicine, behaves as a bifunctional ligand having properties of both the B and C rings of colchicine by making more than one point of attachment with the protein tubulin. PMID- 16700558 TI - pH-Dependent lytic peptides discovered by phage display. AB - Lipid membranes compartmentalize eukaryotic cells and separate the cell interior from the extracellular milieu. So far, studies of peptide and protein interactions with membranes have largely been limited to naturally occurring peptides or to sequences designed on the basis of structural information and biophysical parameters. To expand on these studies, utilizing a system with minimal assumptions, we used phage-display technology to identify 12 amino acid long peptides that bind to liposomes at pH 5.0 but not at pH 7.5. Of the nineteen peptides discovered, three were able to cause leakage of liposome contents. Multivalent presentation of these membrane-active peptides by conjugation onto poly(l-Lysine) enhanced their lytic potential. The secondary structures were analyzed by circular dichroism in aqueous 2,2,2-trifluoroethanol and in buffered aqueous solutions, both in the presence and absence of liposomes. Two of the three lytic peptides show alpha helical profiles, whereas none of the nonlytic peptides formed stable secondary structures. The diverse characteristics of the peptides identified in this study demonstrate that phage-displayed peptide library screens on lipid membranes result in the discovery of nonclassical membrane-active peptides, whose study will provide novel insights into peptide membrane interactions. PMID- 16700559 TI - Dominant negative mutants of transducin-alpha that block activated receptor. AB - Mutations counterpart to dominant negative RasSer17Asn in the alpha-subunits of heterotrimeric G-proteins are known to also produce dominant negative effects. The mechanism of these mutations remains poorly understood. Here, we examined the effects and mechanism of the Ser43Cys and Ser43Asn mutants of transducin-like chimeric Gtalpha* in the visual signaling system. Our analysis showed that both mutants have reduced affinity for GDP and are likely to exist in an empty-or partially occupied-pocket state. S43C and S43N retained the ability to interact with Gtbetagamma and, as heterotrimeric proteins, bind to photoexcited rhodopsin (R*). The interaction with R* is unproductive as the mutants failed to bind GTPgammaS and become activated. S43C and S43N inhibited R*-dependent activation of Gtalpha* and Gtalpha, apparently by blocking R*. Finally, both Gtalpha* mutants lacked interaction with the gamma-subunit of PDE6, an effector protein in phototransduction. These results indicate that the S43C and S43N mutants of Gtalpha* are dominant negative inhibitors that bind and block the activated receptor in a mechanism that parallels that of RasSer17Asn. Dominant negative mutants of Gtalpha sequestering R*, such as S43C and S43N, may become useful instruments in probing the mechanisms of visual dysfunctions caused by abnormal phototransduction signaling. PMID- 16700560 TI - The nickel site of Bacillus pasteurii UreE, a urease metallo-chaperone, as revealed by metal-binding studies and X-ray absorption spectroscopy. AB - UreE is a homodimeric metallo-chaperone that assists the insertion of Ni(2+) ions in the active site of urease. The crystal structures of UreE from Bacillus pasteurii and Klebsiella aerogenes have been determined, but the details of the nickel-binding site were not elucidated due to solid-state effects that caused disorder in a key portion of the protein. A complementary approach to this problem is described here. Titrations of wild-type Bacillus pasteurii UreE (BpUreE) with Ni(2+), followed by metal ion quantitative analysis using inductively coupled plasma optical emission spectrometry (ICP-OES), established the binding of 2 Ni(2+) ions to the functional dimer, with an overall dissociation constant K(D) = 35 microM. To establish the nature, the number, and the geometry of the ligands around the Ni(2+) ions in BpUreE-Ni(2), X-ray absorption spectroscopy data were collected and analyzed using an approach that combines ab initio extended X-ray absorption fine structure (EXAFS) calculations with a systematic search of several possible coordination geometries, using the Simplex algorithm. This analysis indicated the presence of Ni(2+) ions in octahedral coordination geometry and an average of two histidine residues and four O/N ligands bound to each metal ion. The fit improved significantly with the incorporation, in the model, of a Ni-O-Ni moiety, suggesting the presence of a hydroxide-bridged dinuclear cluster in the Ni-loaded BpUreE. These results were interpreted using two possible models. One model involves the presence of two identical metal sites binding Ni(2+) with negative cooperativity, with each metal ion bound to the conserved His(100) as well as to either His(145) or His(147) from each monomer, residues found largely conserved at the C-terminal. The alternative model comprises the presence of two different binding sites featuring different affinity for Ni(2+). This latter model would involve the presence of a dinuclear metallic core, with one Ni(2+) ion bound to one His(100) from each monomer, and the second Ni(2+) ion bound to a pair of either His(145) or His(147). The arguments in favor of one model as compared to the other are discussed on the basis of the available biochemical data. PMID- 16700561 TI - Mutations in PRP43 that uncouple RNA-dependent NTPase activity and pre-mRNA splicing function. AB - Saccharomyces cerevisiae Prp43 is a DEAH-box RNA-dependent ATPase that catalyzes the release of excised lariat intron from the mRNA spliceosome. Previous studies identified mutations in Prp43 motifs I, II, and VI that were lethal in vivo and ablated ATP hydrolysis in vitro. Such Prp43 mutants exerted dominant-negative growth phenotypes when expressed in wild type cells and blocked intron release in vitro when added to yeast splicing extracts. Here, we assessed the effects of alanine and conservative substitutions at conserved residues in motifs Ia ((146)TQPRRVAA(153)), IV ((307)LLFLTG(312)), and V ((376)TNIAETSLT(384)) and thereby identified Arg150 (motif Ia), Phe309 (motif IV), Thr376, Leu383, and Thr384 (motif V) as being important for Prp43 function in vivo. Motif V mutations T376V, T384A, and T384V were lethal and dominant negative in vivo, and the mutant proteins inhibited lariat release in vitro. The T384A and T384V proteins were proficient for ATP hydrolysis, suggesting that ATPase activity is necessary, but not sufficient, for Prp43 function. We report that Prp43 hydrolyzes all common NTPs and dNTPs and unwinds short 5'/3' tailed RNA/DNA duplexes in an ATP dependent fashion. Optimal ATP hydrolysis requires an RNA cofactor of >or=20 nt. Prp43 is largely indifferent to mutations in its C-terminal segment, which is conserved in the DEAH-box splicing factors Prp2, Prp16, and Prp22. PMID- 16700562 TI - DNA ligase I is an in vivo substrate of DNA-dependent protein kinase and is activated by phosphorylation in response to DNA double-strand breaks. AB - DNA-dependent protein kinase (DNA-PK) phosphorylates several cellular proteins in vitro, but its cellular function and natural substrate(s) in vivo are not established. We reported activation of DNA ligase in cultured normal human epidermal keratinocytes (NHEK) on exposure to the DNA-damaging compound bis-(2 chloroethyl) sulfide. The activated enzyme was identified as DNA ligase I, and this activation was attributed to phosphorylation of the enzyme. Here, we show that the phosphorylation is mediated by DNA-PK and that DNA ligase I is one of its natural substrates in vivo. DNA ligase I phosphorylation-cum-activation is a response specific to DNA double-strand breaks. We also demonstrate that affinity purified inactive DNA ligase I is phosphorylated and activated in vitro by HeLa Cell DNA-PK confirming the in vivo observations. The findings specify the roles of DNA-PK and DNA ligase I in mammalian DNA double-strand break repair. PMID- 16700563 TI - Deletion of all cysteines in tachyplesin I abolishes hemolytic activity and retains antimicrobial activity and lipopolysaccharide selective binding. AB - Tachyplesin I is a cyclic beta-sheet antimicrobial peptide isolated from the hemocytes of Tachypleus tridentatus. The four cysteine residues in tachyplesin I play a structural role in imparting amphipathicity to the peptide which has been shown to be essential for its activity. We investigated the role of amphipathicity using an analogue of tachyplesin I (TP-I), CDT (KWFRVYRGIYRRR NH(2)), in which all four cysteines were deleted. Like TP-I, CDT shows antimicrobial activity and disrupts Escherichia coli outer membrane and model membranes mimicking bacterial inner membranes at micromolar concentrations. The CDT peptide does not cause hemolysis up to 200 microg/mL while TP-I showed about 10% hemolysis at 100 microg/mL and about 25% hemolysis at 150 microg/mL. Peptide into-lipid titrations under isothermal conditions reveal that the interaction of CDT with lipid membranes is an enthalpy-driven process. Binding assays performed using fluorometry demonstrate that the peptide CDT binds and inserts into only negatively charged membranes. The peptide-induced thermotropic phase transition of MLVs formed of DMPC and the DMPC/DMPG (7:3) mixture suggests specific lipid peptide interactions. The circular dichroism study shows that the peptide exists as an unordered structure in an aqueous buffer and adopts a more ordered beta structure upon binding to negatively charged membrane. The NMR data suggest that CDT binding to negatively charged bilayers induces a change in the lipid headgroup conformation with the lipid headgroup moving out of the bilayer surface toward the water phase, and therefore, a barrel stave mechanism of membrane disruption is unlikely as the peptide is located near the headgroup region of lipids. The lamellar phase (31)P chemical shift spectra observed at various concentrations of the peptide in bilayers suggest that the peptide may function neither via fragmentation of bilayers nor by promoting nonlamellar structures. NMR and fluorescence data suggest that the presence of cholesterol inhibits the peptide binding to the bilayers. These properties help to explain that cysteine residues may not contribute to antimicrobial activity and that the loss of hemolytic activity is due to lack of hydrophobicity and amphipathicity. PMID- 16700566 TI - Phase separation of a mixed self-assembled monolayer prepared via a stepwise method. AB - Self-assembled monolayers (SAMs), a molecular-level assembly that forms spontaneously, provide a vehicle for investigating specific interactions at interfaces. This is particularly true for mixed SAMs that are composed of organosilanes with different chain lengths and/or chemical functionalities because they offer an adjustable surface for constructing 3D structures containing a variety of moieties. We recently observed that coadsorbed monolayers with different organosilanes on a Si wafer were separated into several tens or hundreds of nanometer domains that were rich in individual components. Several organosilanes, such as octadecyltrichlorosilane (OTS), octadecyltrimethoxysilane (OTMS), (3-mercaptopropyl)trimethoxysilane (MPTMS), and (3 aminopropryl)trimethoxysilane (APTMS), were used for regional separation. In this study, we propose a stepwise deposition method, namely, the deposition of a second siliane on a SAM substrate that creates intentional defects in the first silane. The surface morphologies were adjusted by the deposition sequence and immersion time of the silanes. As a result, a mixed SAM prepared by the proposed method showed effectively functionalized films compared to that prepared by the one-step method. PMID- 16700564 TI - Conformational dynamics of loop 262-274 in G- and F-actin. AB - According to the original Holmes model of F-actin structure, the hydrophobic loop 262-274 stabilizes the actin filament by inserting into a pocket formed at the interface between two protomers on the opposing strand. Using a yeast actin triple mutant, L180C/L269C/C374A [(LC)(2)CA], we showed previously that locking the hydrophobic loop to the G-actin surface by a disulfide bridge prevents filament formation. We report here that the hydrophobic loop is mobile in F- as well as in G-actin, fluctuating between the extended and parked conformations. Copper-catalyzed, brief air oxidation of (LC)(2)CA F-actin on electron microscopy grids resulted in the severing of thin filaments and their conversion to amorphous aggregates. Disulfide, bis(methanethiosulfonate) (MTS), and dibromobimane (DBB) cross-linking reactions proceeded in solution at a faster rate with G- than with F-actin. Cross-linking of C180 to C269 by DBB (4.4 A) in either G- or F-actin resulted in shorter and less stable filaments. The cross linking with a longer MTS-6 reagent (9.6 A) did not impair actin polymerization or filament structure. Myosin subfragment 1 (S1) and tropomyosin inhibited the disulfide cross-linking of phalloidin-stabilized F-actin. Electron paramagnetic resonance measurements with nitroxide spin-labeled actin revealed strong spin spin coupling and a similar mean interspin distance ( approximately 10 A) in G- and in F-actin, with a broader distance distribution in G-actin. These results show loop 262-274 fluctuations in G- and F-actin and correlate loop dynamics with actin filament formation and stability. PMID- 16700567 TI - Highly sensitive biomolecular fluorescence detection using nanoscale ZnO platforms. AB - Fluorescence detection is currently one of the most widely used methods in the areas of basic biological research, biotechnology, cellular imaging, medical testing, and drug discovery. Using model protein and nucleic acid systems, we demonstrate that engineered nanoscale zinc oxide structures can significantly enhance the detection capability of biomolecular fluorescence. Without any chemical or biological amplification processes, nanoscale zinc oxide platforms enabled increased fluorescence detection of these biomolecules when compared to other commonly used substrates such as glass, quartz, polymer, and silicon. The use of zinc oxide nanorods as fluorescence enhancing substrates in our biomolecular detection permitted sub-picomolar and attomolar detection sensitivity of proteins and DNA, respectively, when using a conventional fluorescence microscope. This ultrasensitive detection was due to the presence of ZnO nanomaterials which contributed greatly to the increased signal-to-noise ratio of biomolecular fluorescence. We also demonstrate the easy integration potential of zinc oxide nanorods into periodically patterned nanoplatforms which, in turn, will promote the assembly and fabrication of these materials into multiplexed, high-throughput, optical sensor arrays. These zinc oxide nanoplatforms will be extremely beneficial in accomplishing highly sensitive and specific detection of biological samples involving nucleic acids, proteins and cells, particularly under detection environments involving extremely small sample volumes of ultratrace-level concentrations. PMID- 16700568 TI - Conducting block copolymer for simple micro- to nanopatterns. AB - To use conducting polymers as substitutes for metals and conventional semiconductors in device fabrication, a cost-effective process for the reproducible deposition of the conducting polymers is needed. In this letter, we report a simple solution casting method for the fabrication of micro- to nanopatterns using the conducting block copolymer, poly(thiophene-block ethyleneoxide), which shows rectifying characteristics dependent on the pattern width. PMID- 16700569 TI - Orientation of native cellulose in an electric field. AB - Native cellulose has been oriented in an ac electric field at both the macroscopic and colloidal level. Ramie fiber fragments suspended in chloroform have been shown to point along the field. Cellulose microcrystal suspensions in cyclohexane have also been allowed to evaporate in an electric field and have exhibited a high degree of orientation when further examined by TEM and electron diffraction. Similarly, cellulose whisker suspensions showed increasing birefringence with increasing field strength and displayed interference Newton colors that saturated at around 2000 V cm(-)(1). A high degree of order of this suspension was also obtained by evaluating the induced birefringence with color charts. PMID- 16700570 TI - Brewster angle microscopy study of poly(epsilon-caprolactone) crystal growth in Langmuir films at the air/water interface. AB - Surface pressure-induced crystallization of poly(epsilon-caprolactone) (PCL) from a metastable region of the surface pressure-area per monomer (Pi-A) isotherm in Langmuir monolayers at the air/water (A/W) interface has been captured in real time by Brewster angle microscopy (BAM). Morphological features of PCL crystals grown in Langmuir films during the compression process exhibit four fully developed faces and two distorted faces. During expansion of the crystallized film, polymer chains slowly detach from the crystalline domains and diffuse back into the monolayer as the crystals "melt". Typical diffusion-controlled morphologies are revealed by BAM during the melting process as the secondary dendrites melt away faster, that is, at a higher surface pressure than the principal axes. Electron diffraction on Langmuir-Schaefer films suggests that the lamellar crystals are oriented with the polymer chain axes perpendicular to the substrate surface, while atomic force microscopy reveals a crystal thickness of approximately 7.6 nm. PMID- 16700571 TI - Direct observation of internal fluidity in a water droplet during sliding on hydrophobic surfaces. AB - In the current study, we used a high-speed camera system with particle image velocimetry to observe the internal fluidity of water droplets during sliding. The droplets' velocity during sliding was controlled by slipping and rolling motions. On the superhydrophobic surface, with a contact angle of 150 degrees, the droplet fell at high velocity by slipping. However, on a normal hydrophobic surface whose water contact angle was around 100 degrees, both slipping and rolling controlled the droplet's velocity during sliding. In addition, the advancing velocity might be large when the slip velocity is large and the contact area is small. PMID- 16700572 TI - Polymeric vesicle permeability: a facile chemical assay. AB - We present a simple method to characterize vesicles and determine, at the same time, the membrane permeability to specific molecules. The method is based on encapsulating highly hydrophilic 3,3',3' '-phosphinidynetris-benzenesulfonic acid (PH) into vesicles and subsequently monitoring its reaction with 5,5'-dithiobis-2 nitrobenzoic acid (DTNB). We tested the method by measuring the membrane permeability of vesicles formed from a series of poly(ethylene oxide)-co polybutylene oxide (EB) copolymers and egg yolk phosphatidylcholine. We found that the experimental data are in good agreement with calculations based on Fick's first law. We therefore quantified the DTNB permeability across vesicle membranes, finding that polymeric EB membranes have a more selective permeability toward polar molecules compared to phospholipids membranes. PMID- 16700573 TI - Two-dimensional fluidics based on differential lyophobicity and gravity. AB - We have prepared planar fluidics devices using binary chemical patterns consisting of hydrophobic "roads" on which water droplets slide easily and more hydrophobic "curbs" that direct droplet motion. Contact angle and contact angle hysteresis both control the motion of liquid droplets on surfaces. The difference between the advancing contact angles of the two regions prevents the liquid from crossing the interface between them. The low hysteresis of the roads allows facile movement. Gravity (slight tilting of samples) forces droplets to move effortlessly in defined pathways even though the difference in contact angles is not large and both regions are hydrophobic. PMID- 16700574 TI - Superhydrophobic surfaces prepared by microstructuring of silicon using a femtosecond laser. AB - We present a simple method for fabricating superhydrophobic silicon surfaces. The method consists of irradiating silicon wafers with femtosecond laser pulses and then coating the surfaces with a layer of fluoroalkylsilane molecules. The laser irradiation creates a surface morphology that exhibits structure on the micro- and nanoscale. By varying the laser fluence, we can tune the surface morphology and the wetting properties. We measured the static and dynamic contact angles for water and hexadecane on these surfaces. For water, the microstructured silicon surfaces yield contact angles higher than 160 degrees and negligible hysteresis. For hexadecane, the microstructuring leads to a transition from nonwetting to wetting. PMID- 16700575 TI - Thermosensitive properties of poly(amidoamine) dendrimers with peripheral phenylalanine residues. AB - Dendrimers are unique polymers with globular shapes and well-defined structures. We previously prepared poly(amidoamine) (PAMAM) dendrimers having phenylalanine (Phe) residues at every chain end of the dendrimer as efficient gene carriers. In this study, we found that Phe-derivatized PAMAM dendrimers change their water solubility depending on temperature. The dendrimers were soluble in aqueous solutions at low temperatures, but they became water-insoluble at temperatures above a specific threshold, which is termed the lower critical solution temperature (LCST). Although the LCST of Phe-modified dendrimers decreased with increasing dendrimer generation, these dendrimers exhibited an LCST of 20-30 degrees C under physiological conditions. In addition, the LCST of the dendrimers was controlled by introducing isoleucine (Ile) residues at chain ends of dendrimers at varying ratios with respect to Phe residues. The PAMAM dendrimers are known to encapsulate various drug molecules. For these reasons, temperature sensitive dendrimers might be useful as efficient drug carriers with controlled size and temperature-responsive properties. PMID- 16700576 TI - Polystyrene-silica nanocomposite particles via alcoholic dispersion polymerization using a cationic azo initiator. AB - Submicrometer-sized polystyrene-silica nanocomposite particles have been prepared by alcoholic dispersion polymerization of styrene using commercial alcoholic silica sols of 13 or 22 nm diameter as the sole stabilizing agent. The key to the formation of colloidally stable nanocomposite particles is the selection of a cationic azo initiator (use of nonionic or anionic initiators leads either to the formation of silica-stabilized polystyrene latex particles with very low silica contents or to the precipitation of polystyrene, respectively). Neither surface modification of the silica sol nor the addition of surfactant or polymeric stabilizers is required for successful nanocomposite syntheses. The purified polystyrene-silica nanocomposite particles have relatively narrow particle size distributions, with mean diameters ranging from 331 to 464 nm as judged by disk centrifuge photosedimentometry. Thermogravimetric analyses indicated mean silica contents of 13-26 wt. %, depending on the synthesis conditions. Calcination of the polystyrene-silica nanocomposite particles leads to the formation of hollow silica shells, which indicates a well-defined core-shell morphology for the original nanocomposite particles. PMID- 16700577 TI - High-pressure liquid dispersion and fragmentation of flame-made silica agglomerates. AB - The influence of primary particle diameter and the degree of agglomeration of flame-made silica agglomerate suspensions in aqueous solutions is studied by high pressure dispersion (up to 1500 bar) through a nozzle with a 125 microm inner diameter. These particles were produced (4-15 g/h) by oxidation of hexamethyldisiloxane (HMDSO) in a coflow diffusion flame reactor. Their average primary particle size (10-50 nm) and degree of agglomeration were controlled by varying the oxygen and precursor flow rates. The particles were characterized by nitrogen adsorption, electron microscopy, and small-angle X-ray scattering. Hydrodynamic stresses break up soft agglomerates and yield hard agglomerate sizes in the range of 100-180 nm, as characterized by dynamic light scattering. Soft agglomerates exhibited decreasing light scattering diameters with increasing dispersion pressure, while hard agglomerates were insensitive. PMID- 16700578 TI - Control over colloidal aggregation in monolayers of latex particles at the oil water interface. AB - The controlled generation of 2D aggregate networks is studied experimentally using micrometer-sized polystyrene latex particles attached to the oil-water interface. Starting from an initially crystalline monolayer, appropriate combinations of carefully added electrolyte and surfactant enable control over both the fractal dimension and the kinetics of aggregation. Remarkably, the colloidal crystals formed upon first spreading remain stable, even for days, when substantial amounts of electrolyte are added to the aqueous phase. Pressure-area isotherms reveal a slow time evolution of the electrostatic dipole-dipole interaction. When the electrostatic interaction has been sufficiently weakened, aggregation proceeds in well-defined, reproducible manner. The aggregation process is analyzed using quantitative video microscopy. The evolution of the cluster size distribution and its moments is characterized, and static and dynamic scaling exponents are derived to identify the nature of the aggregation process. In the range of concentrations studied here, the kinetics all agree with a "fast", diffusion-limited cluster type of aggregation. However, the fractal dimension strongly depends on the composition of the aqueous subphase. Rather dense structures are found when only electrolyte is used, whereas more open structures are obtained when even small amounts of surfactant are added. It is suggested that this structural dependency is related to the effect of surfactant on the contact angle and its consequences for the anisotropic nature of the capillary interactions. PMID- 16700579 TI - Study on improving the turbidity measurement of the absolute coagulation rate constant. AB - The existing theories dealing with the evaluation of the absolute coagulation rate constant by turbidity measurement were experimentally tested for different particle-sized (radius = a) suspensions at incident wavelengths (lambda) ranging from near-infrared to ultraviolet light. When the size parameter alpha = 2pi a/lambda > 3, the rate constant data from previous theories for fixed-sized particles show significant inconsistencies at different light wavelengths. We attribute this problem to the imperfection of these theories in describing the light scattering from doublets through their evaluation of the extinction cross section. The evaluations of the rate constants by all previous theories become untenable as the size parameter increases and therefore hampers the applicable range of the turbidity measurement. By using the T-matrix method, we present a robust solution for evaluating the extinction cross section of doublets formed in the aggregation. Our experiments show that this new approach is effective in extending the applicability range of the turbidity methodology and increasing measurement accuracy. PMID- 16700580 TI - Interactions of hydrophobically modified polyvinylamine with pluronic triblock copolymer micelles. AB - The heats associated with the addition of triblock copolymers of ethylene oxide and propylene oxide (PEO-PPO-PEO pluronics) to solutions of linear polyvinylamine, with N-substituted pendant octyl groups (HMPVAm), were measured as functions of pH and pluronic structure. The interactions were exothermic with the overall enthalpy decreasing with increasing pH from 5 to 10. Surprisingly, the heat effect increased with increasing pluronic hydrophilicity; however, no enthalpy change was observed in the absence of micelles. The results were quantitatively modeled by assuming two competing processes-micellar dissolution and HMPVAm coating of micelles, preventing dissolution. PMID- 16700581 TI - Restructuring and break-up of two-dimensional aggregates in shear flow. AB - We consider single two-dimensional aggregates, containing glass particles, placed at a water/air interface. We have investigated the critical shear rate for break up of aggregates with different sizes in a simple shear flow. All aggregates break-up nearly at the same shear rate (1.8 +/- 0.2 s(-)(1)) independent of their size. The evolution of the aggregate structure before break-up was also investigated. With increasing shear rate, the aggregates adopt a more circular shape, and the particles order in a more dense, hexagonal structure. A simple theoretical model was developed to explain the experimentally observed break-up. In the model, the aggregate is considered as a solid circular disk that will break near its diameter. The capillary and drag force on the two parts of the aggregate were calculated, and from this force balance, the critical shear rate was found. The model shows a weak size dependence of the critical shear rate for the considered aggregates. This is consistent with the experimental observations. PMID- 16700582 TI - Latex-particle-stabilized emulsions of anti-Bancroft type. AB - Here, we investigate water-in-oil (W/O) emulsions that are stabilized by polystyrene latex particles with sulfate surface groups. The particles, which play the role of emulsifier, are initially contained in the disperse (water) phase. The existence of such emulsions formally contradicts the empirical Bancroft rule. Theoretical considerations predict that the drop diameter has to be inversely proportional to the particle concentration, but should be independent of the volume fraction of water. In addition, there should be a second emulsification regime, in which the drop diameter is determined by the input mechanical energy during the homogenization. The existence of these two regimes has been experimentally confirmed, and the obtained data agree well with the theoretical model. Stable W/O emulsions have been produced with hexadecane and tetradecane, while, in the case of more viscous and polar oils (soybean and silicone oil), the particles enter into the oily phase, and Pickering emulsions cannot be obtained. The formation of stable emulsions demands the presence of a relatively high concentration of electrolyte that lowers the electrostatic barrier to particle adsorption at the oil-water interface. Because the attachment of particles at the drop surfaces represents a kind of coagulation, it turns out that the Schulze-Hardy rule for the critical concentration of coagulation is applicable also to emulsification, which has been confirmed with suspensions containing Na(+), Mg(2+), and Al(3+) counterions. The increase of the particle and electrolyte concentrations and the decrease of the volume fraction of water are other factors that facilitate emulsification in the investigated system. To quantify the combined action of these factors, an experimental stability instability diagram has been obtained. PMID- 16700583 TI - Deposition of DNA-functionalized gold nanospheres into nanoporous surfaces. AB - We report the deposition of DNA-conjugated gold nanospheres into arrays of surface nanopores obtained from hexagonally ordered thin polystyrene-b poly(methyl methacrylate) (PS-b-PMMA) diblock copolymer films on silicon. The deposition occurs spontaneously from aqueous solution and is driven by either electrostatic interactions or specific DNA hybridization events between the DNA nanospheres and the surface nanopores. To mitigate this spontaneous deposition, we have chemically modified the nanopores with either positively charged aminosilanes or oligonucleotide probe sequences. The deposition of DNA nanospheres into the surface nanopores was characterized by atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS). We have observed preferential immobilization of individual DNA nanospheres within the nanopores, based on the size matching between the two entities. The inclusion density and selectivity of DNA nanosphere deposition into the surface nanopores was found to depend predominantly on the methods through which the nanoporous surfaces were prepared and chemically functionalized. PMID- 16700584 TI - Surface-deformation characteristics of uniaxially oriented poly(ethylene terephthalate) film as evaluated from nanoscratch tests with scanning probe microscopy. AB - The surface-deformation characteristics of uniaxially drawn poly(ethylene terephthalate) (PET) film were successfully evaluated with multiline scratch tests using scanning probe microscopy (SPM) on a nanometer scale. The PET film was prepared by compression molding from the melt, followed by quenching in ice water. The obtained amorphous film was drawn uniaxially below its glass transition temperature, and the resultant surface roughness could be reduced to within 5 nm. A multiline scratch with the Si(3)N(4) tip of an SPM on the oriented PET surface was made parallel and perpendicular to the drawing axis under applied loads of 5-30 nN. The perpendicular scratching generated a characteristic periodic pattern on the film surface, but the parallel scratching induced a tearing of the surface. These results suggest that surface-deformation mechanisms were dominated by molecular anisotropy. The surface-deformation properties, as evaluated from scratch-angle dependences on morphological changes on a nanometer scale, were similar to the mechanical properties of the bulk. PMID- 16700585 TI - Mesoscale pincushions, microrings, and microdots prepared by heating and peeling of self-organized honeycomb-patterned films deposited on a solid substrate. AB - We describe here a preparation of pincushion structures with holes, hexagonally arranged microrings, and microdots by simple heating and peeling of self organized honeycomb-patterned films. We have reported that the honeycomb patterned films can be prepared by casting the solution of an amphiphilic polymer and a hydrophobic polymer under humid conditions. When annealing the honeycomb patterned films prepared from an amphiphilic copolymer and poly(bisphenol A carbonate), we obtained a variety of mesoscale structures, depending on the heating temperatures. We revealed that these microstructures were formed by using the phase-separation structures in the self-organized honeycomb-patterned films. These micropatterns can be utilized for the template for microelectrodes, superhydrophobic surfaces, photonic crystals, and as a substrate for tissue engineering. PMID- 16700587 TI - Supramolecular assemblies of a new series of gemini-type schiff base amphiphiles at the air/water interface: in situ coordination, interfacial nanoarchitectures, and spacer effect. AB - Great interest has been devoted to the gemini amphiphiles because of their unique properties. In this article, we report some interesting properties of the interfacial films formed by a series of newly designed gemini amphiphiles containing the Schiff base moiety. This novel series of gemini amphiphiles with their Schiff base headgroups linked by a hydrophobic alkyl spacer (BisSBC18Cn, n = 2, 4, 6, 8, 10) could be spread to form stable monolayers and coordinated with Cu(Ac)(2) in situ in the monolayer. The alkyl spacer in the amphiphiles has a great effect on the regulation of the properties of the Langmuir monolayers. A maximum limiting molecular area was observed for the monolayers of the gemini amphiphile with the spacer length of hexa- or octamethylene groups. Both the monolayers on water and on the aqueous Cu(Ac)(2) subphase were transferred onto solid substrates, and different morphologies were observed for films with different spacers. Nanonail and tapelike morphologies were observed for amphiphile films with shorter spacers (n = 2 and 4) on the water surface. Wormlike morphologies were observed for gemini films with longer spacers of C(8) and C(10) when coordinated with Cu(Ac)(2). An interdigitated layer structure was supposed to form in the multilayer films transferred from water or the aqueous Cu(Ac)(2) subphase. PMID- 16700586 TI - Contact line and contact angle dynamics in superhydrophobic channels. AB - The dynamics of the wetting and movement of a three-phase contact line confined between two superhydrophobic surfaces were studied using a mean-field free-energy lattice Boltzmann model. Principle features of superhydrophobic surfaces, such as trapped vapor/air between rough microstructures, high contact angles, reduced contact angle hysteresis, and low resistance to fluid flow, were all observed. Movement of the three-phase contact line over a well-patterned superhydrophobic surface displays a periodic stick-jump-slip behavior, while the dynamic contact angle changes accordingly from maximum to minimum. Two regimes were found for the flow velocity as a function of surface roughness and can be related directly to the balance between driving force and flow resistance. This work provides a better understanding of dynamic wetting and fluid flow behaviors over superhydrophobic surfaces and hence could be useful in related applications. PMID- 16700588 TI - Interaction forces in thin liquid films stabilized by hydrophobically modified inulin polymeric surfactant. 1. Foam films. AB - Using the interferometric method of Scheludko-Exerowa for investigation of foam films, we have obtained results using a hydrophobically modified inulin polymeric surfactant (INUTEC SP1). Measurements were carried out at constant INUTEC SP1 concentration of 2 x 10(-)(5) mol.dm(-)(3) and at various NaCl concentrations (in the range 1 x 10(-)(4) to 2 mol.dm(-)(3)). At constant capillary pressure of 50 Pa, the film thickness decreased gradually with an increase in NaCl concentration up to 10(-)(2) mol.dm(-)(3) NaCl above which the film thickness remains virtually constant at about 16 nm. This reduction in film thickness with an increase in NaCl concentration is due to the compression of the double layer and at the critical electrolyte concentration (C(el,cr) = 10(-)(2) mol.dm(-)(3)) the electrostatic component of the disjoining pressure is completely screened and the remaining pressure is due to the steric interaction between the adsorbed polymer layers. Disjoining pressure-thickness (Pi-h) isotherms were obtained at C(el) < C(el,cr) (10(-)(4) - 10(-)(3) mol.dm(-)(3)) and C(el) > C(el,cr) (0.5, 1, and 2 mol.dm(-)(3)). In the first case, the disjoining pressure isotherms could be fitted using the classical DLVO theory, Pi = Pi(el) + Pi(vw), and using the constant charge model. At C(el) > C(el,cr), the main repulsion is due to the steric interaction between the polyfructose loops that exist at the air-water interface, i.e., Pi = Pi(st) + Pi(vw). Under these conditions, there is a sharp transition from DLVO to non-DLVO forces. In the latter case, the interaction could be described using the de Gennes' scaling theory. This gave an adsorbed layer thickness of 6.5 nm which is in reasonable agreement with the values obtained at the solid-solution interface. The Pi-h isotherms showed that these foam films are not very stable and they tend to collapse above a critical capillary pressure (of about 1 x 10(3) Pa), and these results could be used to predict the foam stability. PMID- 16700589 TI - Photocatalytic reduction of an azide-terminated self-assembled monolayer using CdS quantum dots. AB - Ordered, tightly packed aryl-azide-terminated, self-assembled monolayers (SAMs) were created on gold substrates from a new disulfide precursor. These monolayers were reduced at least partially in an aqueous environment using approximately 2 nm CdS quantum dots (Qdots) as photocatalysts to give mixed monolayers of arylamine- and aryl azide-terminated species. The CdS photocatalysts were made available for the reaction by exposure of the azide-terminated SAM to Qdots initially in solution or by preadsorption of the CdS nanoparticles on the SAM. In either case, X-ray photoelectron spectroscopy (XPS), grazing angle Fourier transform infrared spectroscopy (FTIR), and contact angle measurements were used to show the occurrence of the photocatalytic reduction. As further evidence for the presence of arylamine-terminated thiolate in the reduced SAM, these arylamine groups were successfully tagged with fluorescein isothiocyanate (FITC). The use of Qdot photocatalysts to functionalize surfaces may lead to a means to pattern surfaces at the nanoscale. PMID- 16700590 TI - Physical properties of nanobubbles on hydrophobic surfaces in water and aqueous solutions. AB - In recent years there has been an accumulation of evidence for the existence of nanobubbles on hydrophobic surfaces in water, despite predictions that such small bubbles should rapidly dissolve because of the high internal pressure associated with the interfacial curvature and the resulting increase in gas solubility. Nanobubbles are of interest among surface scientists because of their potential importance in the long-range hydrophobic attraction, microfluidics, and adsorption at hydrophobic surfaces. Here we employ recently developed techniques designed to induce nanobubbles, coupled with high-resolution tapping-mode atomic force microscopy (TM-AFM) to measure some of the physical properties of nanobubbles in a reliable and repeatable manner. We have reproduced the earlier findings reported by Hu and co-workers. We have also studied the effect of a wide range of solutes on the stability and morphology of these deliberately formed nanobubbles, including monovalent and multivalent salts, cationic, anionic, and nonionic surfactants, as well as solution pH. The measured physical properties of these nanobubbles are in broad agreement with those of macroscopic bubbles, with one notable exception: the contact angle. The nanobubble contact angle (measured through the denser aqueous phase) was found to be much larger than the macroscopic contact angle on the same substrate. The larger contact angle results in a larger radius of curvature and a commensurate decrease in the Laplace pressure. These findings provide further evidence that nanobubbles can be formed in water under some conditions. Once formed, these nanobubbles remain on hydrophobic surfaces for hours, and this apparent stability still remains a well recognized mystery. The implications for sample preparation in surface science and in surface chemistry are discussed. PMID- 16700591 TI - Swelling and deswelling of adsorbed microgel monolayers triggered by changes in temperature, pH, and electrolyte concentration. AB - The formation and characterization of close-packed monolayers of negative, poly(N isopropylacrylamide)-based microgel particles onto positively charged silicon wafers is described. The silicon wafers were rendered positive by first oxidizing their surface to silica and then adsorbing a layer of poly(ethyleneimine). The thickness of the deposited microgel monolayers (under aqueous conditions) has been determined by spectroscopic ellipsometry as a function of temperature (20-60 degrees C), pH (3-8), and added NaCl concentration (0-1 M). No actual desorption of the microgel particles was evident on changing the conditions, but a swelling/deswelling transition was observed around 32 degrees C. The thickness of the monolayer has been compared with the hydrodynamic diameter of the free microgel particles, dispersed in aqueous solution. For the poly(N isopropylacrylamide) microgel particles, without any bulk ionisable comonomer groups present, the temperature dependence of the ellipsometric thickness of the monolayer mirrors closely that of the hydrodynamic diameter of the free particles. When ionizable (-COOH) groups are introduced into the microgel particles, however, this correspondence is largely lost because the microgel particles forming the deposited monolayer now contract strongly onto the oppositely charged substrate surface. PMID- 16700592 TI - Effect of peptide length on the interaction between consensus peptides and DOPC/DOPA bilayers. AB - The effect of peptide length and electrostatics on the interaction between Cardin motif peptides and lipid membranes was investigated for (AKKARA)(n) (n = 1-4) and (ARKAAKKA)(n) (n = 1-3) peptides (A, K, and R refer to alanine, lysine, and arginine, respectively) by fluorescence spectroscopy, circular dichroism, ellipsometry, z potential, and photon correlation spectroscopy measurements. The effect of the peptides regarding leakage induction of both zwitterionic and anionic liposomes increased with increasing peptide length, as did the peptide induced killing of Enterococcus faecalis and Bacillus subtilis bacteria. The peptides, characterized by a random coil conformation both in buffer and when attached to the liposomes (helix content less than 20%), displayed an increased adsorption with increasing peptide length, and plateau adsorption for the longest peptides corresponded to 1 peptide per 65 and 17 lipid molecules for zwitterionic and anionic membranes, respectively. Control experiments with uncharged peptide analogues as well as experiments at high excess electrolyte concentration showed that peptide charges are important both for peptide adsorption and leakage induction. These observations, together with observations of the liposome z potential at different peptide additions as well as a comparison between the results for zwitterionic and anionic liposomes, suggest that electrostatically affected local packing effects are crucial for the action of these peptides, although pore formation such as that observed for many AMPs cannot be excluded at present. PMID- 16700593 TI - Self-organization of low-symmetry adjacent-type metallophthalocyanines having branched alkyl chains. AB - Low-symmetry, adjacent-type metallophthalocyanines 1 and 2 with four branched alkyl chains on one side and a chiral bridging segment on the other were synthesized, and their self-organization properties were investigated. The synthesized adjacent-type phthalocyanines were liquid-crystalline and exhibited a phase transition from the crystalline phase to the mesophase below room temperature. X-ray diffraction indicated that the molecules are stacked in one dimensional columnar aggregates with a hexagonal arrangement. The self organization behavior of zinc complex 1 and cobalt complex 2 was also investigated with a monolayer experiment at the air-water interface. The adjacent type phthalocyanines formed a stable monolayer at the air-water interface, and the monolayers could be transferred onto quartz substrates by a Y-type deposition. UV-vis, XRD, and CD measurements for the resulting Langmuir-Blodgett films indicated that 1 and 2 had different molecular orientations. PMID- 16700594 TI - Properties of mixed lipid monolayers assembled on hydrophobic surfaces through vesicle adsorption. AB - Supported lipid films are becoming increasingly important tools for the study of membrane protein function because of the availability of high-sensitivity surface analytical and patterning techniques. In this study, we have characterized the physical chemical properties of lipid films assembled on hydrophobic surfaces through the spontaneous adsorption of large unilamellar lipid vesicles composed of dioleoylphosphatidylglycerol (DOPG) and dioleoylphosphatidylcholine (DOPC). The density of the lipid films was measured with surface plasmon resonance spectroscopy as the lipid composition of the vesicles and ionic concentration were varied. As expected, monolayer films were formed, but the density of the monolayers was found to be weakly dependent on the lipid composition of the vesicles and strongly dependent on the ionic concentration of the solution in contact with the monolayer. Atomic force microscopy (AFM) images of the lipid films indicate that they are composed of a homogeneous monolayer. Surface force measurements were used to determine the surface charge and DOPG density of the monolayers. The DOPG content of the films was found to be weakly dependent on the DOPG composition of the vesicles and strongly dependent on the salt concentration of the environment. A model has been developed to describe the behavior of the lipid composition of the films in terms of the hydrophobic, electrostatic, and steric forces acting on the lipid monolayer on the hydrophobic surface. PMID- 16700595 TI - Adsorption of myoglobin onto porous zirconium phosphate and zirconium benzenephosphonate obtained with template synthesis. AB - Porous zirconium phosphate (P-ZrP) and zirconium benzenephosphonate (P-ZrBP) were prepared in the presence of an anionic surfactant acting as a template. Poorly crystalline materials with a P/Zr molar ratio equal to 2 and having a relatively high surface area and micro/mesoporosity have been obtained. The interaction of myoglobin with the two types of surfaces, the hydrophobic P-ZrBP and the hydrophilic P-ZrP, was investigated, and the adsorption isotherms were determined at different pH and temperature values. A model was proposed for the mechanism of the interaction of the protein with the surface based on the shape of the adsorption isotherm and the physical-chemical properties of myoglobin. The pH has been found to be an important parameter for determining the maximum adsorption capacity of P-ZrBP and P-ZrP for myoglobin molecules because of the changes that occur in the type and net charge of the protein surface as the pH of the medium changes. Protein binding affinity and capacity increase when the temperature is increased. This phenomenon occurs because myoglobin varies its conformation at high temperature with an increase in the exposed hydrophobic region. This process causes a stronger hydrophobic interaction between the protein and the adsorbent and reduces the repulsion between the adsorbed molecules. Studies on the activities of the obtained biocomposites are in progress. PMID- 16700596 TI - Glucose-responsive polyelectrolyte capsules. AB - Phenylboronic acids are known to form covalent complexes with polyol compounds such as glucose. A novel polyelectrolyte, containing phenylboronic acid as a glucose-sensitive moiety, has been synthesized and used for the fabrication of glucose-sensitive hollow polyelectrolyte capsules using the layer-by-layer technique. The response to glucose was observed as a rather fast dissolution of the capsules when brought into contact with a glucose-containing medium. These polyelectrolyte capsules are the first polyelectrolyte capsules able to respond to a stimulus that can be provided by the human body (i.e., an increase in glucose concentration). Therefore, the concept we present has promising applications in the biomedical field for the controlled delivery of insulin. PMID- 16700597 TI - Anti-biofouling properties of comblike block copolymers with amphiphilic side chains. AB - Surfaces of novel block copolymers with amphiphilic side chains were studied for their ability to influence the adhesion of marine organisms. The surface-active polymer, obtained by grafting fluorinated molecules with hydrophobic and hydrophilic blocks to a block copolymer precursor, showed interesting bioadhesion properties. Two different algal species, one of which adhered strongly to hydrophobic surfaces, and the other, to hydrophilic surfaces, showed notably weak adhesion to the amphiphilic surfaces. Both organisms are known to secrete adhesive macromolecules, with apparently different wetting characteristics, to attach to underwater surfaces. The ability of the amphiphilic surface to undergo an environment-dependent transformation in surface chemistry when in contact with the extracellular polymeric substances is a possible reason for its antifouling nature. Near-edge X-ray absorption fine structure spectroscopy (NEXAFS) was used, in a new approach based on angle-resolved X-ray photoelectron spectroscopy (XPS), to determine the variation in chemical composition within the top few nanometers of the surface and also to study the surface segregation of the amphiphilic block. A mathematical model to extract depth-profile information from the normalized NEXAFS partial electron yield is developed. PMID- 16700598 TI - Nanotopographic control of cytoskeletal organization. AB - Growth of 3T3-L1 preadipocytes on a nanoscalar poly(ethylene terephthalate) (PET) surface produced an absence of the intracellular stress fibers characteristic of cell growth on "normal" planar surfaces. This phenomenon was consistently observed from time zero throughout 3 days of culture and was accompanied by changes in paxillin expression along with an approximately 50% decrease in the number of adherent cells in response to 500 dynes/cm(2) of shear stress. This suggests that the cytoskeleton in cells adherent to nanofibrillar surfaces does indeed form, but at a smaller, more difficult to observe scale. We propose a novel mechanism by which the growth and clustering of integrin-associated focal adhesions on surface nanofibrils regulates cytoskeletal development. The width of the extracellular matrix contacts is constrained by the width of the nanofibrils and the absence of any surface between them. The limited dimensions of these point contacts then constrain receptor polymerization and the associated aggregation of actin filaments. The existence of a topographic mechanism leading to growth-limited integrin clustering is hypothesized. PMID- 16700599 TI - Curvature-modulated phase separation in lipid bilayer membranes. AB - Cellular membranes exhibit a variety of controlled curvatures, with filopodia, microvilli, and mitotic cleavage furrows being only a few of many examples. Coupling between local curvature and chemical composition in membranes could provide a means of mechanically controlling the spatial organization of membrane components. Although this concept has surfaced repeatedly over the years, controlled experimental investigations have proven elusive. Here, we introduce an experimental platform, in which microfabricated surfaces impose specific curvature patterns onto lipid bilayers, that allows quantification of mechanochemical couplings in membranes. We find that, beyond a critical curvature value, membrane geometry governs the spatial ordering of phase-separated domain structures in membranes composed of cholesterol and phospholipids. The curvature controlled ordering, a consequence of the distinct mechanical properties of the lipid phases, makes possible a determination of the bending rigidity difference between cholesterol-rich and cholesterol-poor lipid domains. These observations point to a strong coupling between mechanical bending and chemical organization that should have wide-reaching consequences for biological membranes. Curvature mediated patterning may also be useful in controlling complex fluids other than biomembranes. PMID- 16700600 TI - Directed immobilization of protein-coated nanospheres to nanometer-scale patterns fabricated by electron beam lithography of poly(ethylene glycol) self-assembled monolayers. AB - Controlling the spatial organization of biomolecules on solid supports with high resolution is important for a wide range of scientific and technological problems. Here we report a study of electron beam lithography (EBL) patterning of a self-assembled monolayer (SAM) of the amide-containing poly(ethylene glycol) (PEG) thiol CH(3)O(CH(2)CH(2)O)(17)NHCO(CH(2))(2)SH on Au and demonstrate the patterning of biomolecular features with dimensions approaching 40 nm. The electron dose dependence of feature size and pattern resolution is studied in detail by atomic force microscopy (AFM), which reveals two distinct patterning mechanisms. At low doses, the pattern formation occurs by SAM ablation in a self developing process where the feature size is directly dose-dependent. At higher doses, electron beam-induced deposition of material, so-called contamination writing, is seen in the ablated areas of the SAM. The balance between these two mechanisms is shown to depend on the geometry of the pattern. The patterned SAMs were backfilled with fluorescent 40-nm spheres coated with NeutrAvidin. These protein-coated spheres adhered to exposed areas in the SAM with high selectivity. This direct writing approach for patterning bioactive surfaces is a fast and efficient way to produce patterns with a resolution approaching that of single proteins. PMID- 16700601 TI - Adhesion forces between protein layers studied by means of atomic force microscopy. AB - Adhesion forces between different protein layers adsorbed on different substrates in aqueous media have been measured by means of an atomic force microscope using the colloid probe technique. The effects of the loading force, the salt concentration and pH of the medium, and the electrolyte type on the strength, the pull-off distance, and the separation energy of such adhesion forces have been analyzed in depth. Two very different proteins (bovine serum albumin and apoferritin) and two dissimilar substrates (silica and polystyrene) were used in the experiments. The results clearly point out a very important contribution of the electrostatic interactions in the adhesion between protein layers. PMID- 16700602 TI - Changes in adsorbed fibrinogen upon conversion to fibrin. AB - The conversion of adsorbed fibrinogen to fibrin in the presence of the enzyme thrombin was studied using surface plasmon resonance (SPR), a quartz crystal microbalance (QCM), sum frequency generation (SFG), atomic force microscopy (AFM), and an elutability assay. Exposure of adsorbed fibrinogen to thrombin resulted in a mass loss at the surface consistent with fibrinopeptide release and conversion to fibrin. Changes in hydration upon conversion of adsorbed fibrinogen to fibrin were determined from comparisons of acoustic (QCM) and optical (SPR) mass adsorption data. Conversion to fibrin also resulted in the adsorbed layer becoming more strongly bound to the surface and more compact. The elutability of adsorbed fibrinogen by Triton X-100, studied with SPR, decreased from 90 +/- 5 to 6 +/- 2% after conversion to fibrin. The height of the adsorbed monolayer, as determined by AFM, decreased from 5.5 +/- 2.2 to 1.7 +/- 0.8 nm. We conclude that thrombin-catalyzed fibrinopeptide release triggers significant changes in fibrinogen conformation beyond peptide cleavage. PMID- 16700603 TI - Magnetically and biologically active bead-patterned hydrogels. AB - We present a new approach to the direct patterning of biologically and magnetically active microbeads in nonbiofouling polymer scaffolds for use in microfluidic devices. Briefly, the process involves treatment of a glass substrate, conformal contact bonding of a PDMS microchannel on the substrate, filling of the channel with beads and prepolymer solution, and UV-initiated photopolymerization of a mask-defined pattern using a standard inverted microscope. This versatile and simple method allows for the rapid fabrication of dispersed or packed bead patterns in poly(ethylene glycol) (PEG) hydrogels that are covalently linked to glass surfaces. By exploiting the relative opacity of the microbeads used, we are able to create both partially exposed and fully encapsulated bead patterns. To demonstrate the utility of this new technology, we separated magnetic bead-bound B lymphocytes from T lymphocytes on a PEG encapsulated magnetic filtration platform and also captured B cells directly on patterned, protein-decorated beads in a flow-through microfluidic device. Beyond cell sorting, the accurate patterning of industrially standardized, chemically diverse microbeads may have significant implications for microchip-based analyte detection. PMID- 16700604 TI - Electrostatic model for mixed cationic-zwitterionic lipid bilayers. AB - The current interest in mixed cationic-zwitterionic lipid membranes derives from their potential use as transfer vectors in nonviral gene therapy. Mixed cationic zwitterionic lipid membranes have a number of structural properties that are distinct from the corresponding anionic-zwitterionic lipid membranes. As known from experiment and reproduced by computer simulations, the average cross sectional area per lipid changes nonmonotonically with the mole fraction of the charged lipid, passing through a minimum at a roughly equimolar mixture. At the same time, the average orientation of the zwitterionic headgroup dipoles changes from more parallel to the membrane plane to more perpendicular. We suggest a simple mean-field model that reveals the physical mechanisms underlying the observed structural properties. To backup the mean-field calculations, we have also performed Monte Carlo simulations. Our model extends Poisson-Boltzmann theory to include (besides the cationic headgroup charges) the individual charges of the zwitterionic lipid headgroups. We model these charges to be arranged as dipoles of fixed length with rotational degrees of freedom. Our model includes, in a phenomenological manner, the changes in steric headgroup interactions upon reorientation of the zwitterionic headgroups. Our numerical results suggest that two different mechanisms contribute to the observed structural properties: one involves the lateral electrostatic pressure and the other the zwitterionic headgroup orientation, the latter modifying steric headgroup interactions. The two mechanisms operate in parallel as they both originate in the electrostatic properties of the involved lipids. We have also applied our model to a mixed anionic-zwitterionic lipid membrane for which neither a significant headgroup reorientation nor a nonmonotonic change in the average lateral cross-sectional area is predicted. PMID- 16700605 TI - Water distribution in multilayers of weak polyelectrolytes. AB - The water localization in thin polyelectrolyte multilayers assembled from poly(acrylic acid) and poly(allylamine hydrochloride) was investigated with neutron reflectivity in an atmosphere of controlled humidity and with bulk water. Water was found to be distributed asymmetrically within the multilayer and to localize preferentially at the polymer surface. The diffusion of water into the multilayer did not completely penetrate to the substrate, but instead there appeared to be an exclusion zone near the Si substrate. These results help to explain previous observations of anomalous water transport kinetics in weak polyelectrolyte systems. PMID- 16700606 TI - Organic-inorganic hybrid nanoparticles: surface characteristics and interactions with a polyester resin. AB - Organic-inorganic hybrid nanoparticles, derived from silica precursors with different organic functionalities (methyl, ethyl, vinyl, and phenyl) synthesized via a modified Stober method have been investigated. These particles are intended as modifiers for polymers and polymer matrix composites. Therefore, the characteristics of a polyester matrix have also been determined, and the likely interactions with the particles have been proposed. Particles have been characterized using inverse gas chromatography (IGC), X-ray photoelectron spectroscopy (XPS), and infrared spectroscopy (FT-IR). The particles show two different sets of characteristics, with methyl, ethyl, and vinyl modified silicas showing one type of behavior and the phenyl modified silica behaving rather differently. The methyl, ethyl, and vinyl groups exhibit the appearance of uniform coverage, as they are comparatively small and tightly packed, which will prevent interaction of matrix resin with retained silanol groups. The phenyl group, which is comparatively large, is not able to pack as closely, which results in a reduction of the presence and availability of silanol groups, compared to an unmodified fumed silica, but not complete inaccessibility as far as the matrix resin is concerned. PMID- 16700607 TI - Electron beam irradiation of poly(vinyl methyl ether) films. 2. Temperature dependent swelling behavior. AB - Temperature-sensitive hydrogel films were synthesized by electron beam irradiation of poly(vinyl methyl ether) (PVME) on silicon (Si/SiO(2)) substrates and gold (Au) coated glass slides. The temperature-dependent swelling behavior of the films in aqueous solution was characterized by in situ spectroscopic ellipsometry and a combination of surface plasmon resonance (SPR) and optical waveguide spectroscopy (OWS). The results of both techniques are compared. The suitability of both techniques for the characterization of the swelling behavior of thin hydrogel films is demonstrated. The volume swelling degree in the swollen state decreases with increasing radiation dose D. This is explained by the fact that the number of formed polymeric radicals, and hence cross-linking density, increases with D. Above the phase-transition temperature, the swelling degrees were independent of D, slightly above 1. The swelling/deswelling process was fully reversible and is mainly directed perpendicular to the substrate surface. The phase-transition temperature was determined to be T(cr) approximately 33 degrees C. However, T(cr) slightly decreases with increasing D and increasing film thickness d. PMID- 16700608 TI - Improved CO oxidation activity in the presence and absence of hydrogen over cluster-derived PtFe/SiO2 catalysts. AB - The catalytic performance of cluster-derived PtFe/SiO(2) bimetallic catalysts for the oxidation of CO has been examined in the absence and presence of H(2) (PROX) and compared to that of Pt/SiO(2). PtFe(2)/SiO(2) and Pt(5)Fe(2)/SiO(2) samples were prepared from PtFe(2)(COD)(CO)(8) and Pt(5)Fe(2)(COD)(2)(CO)(12) organometallic cluster precursors, respectively. FTIR data indicate that both clusters can be deposited intact on the SiO(2) support. The clusters remained weakly bonded to the SiO(2) surface and could be extracted with CH(2)Cl(2) without any significant changes in their structure. Subsequent heating in H(2) led to complete decarbonylation of the supported clusters at approximately 350 degrees C and the formation of Pt-Fe nanoparticles with sizes in the 1-2 nm range, as indicated by HRTEM imaging. A few larger nanoparticles enriched in Pt were also observed, indicating that a small fraction of the deposited clusters were segregated to the individual components following the hydrogen treatment. A higher degree of metal dispersion and more homogeneous mixing of the two metals were observed during HRTEM/XEDS analysis with the cluster-derived samples, as compared to a PtFe/SiO(2) catalyst prepared through a conventional impregnation route. Furthermore, the cluster-derived PtFe(2)/SiO(2) and Pt(5)Fe(2)/SiO(2) samples were more active than Pt/SiO(2) and the conventionally prepared PtFe/SiO(2) sample for the oxidation of CO in air. However, substantial deactivation was also observed, indicating that the properties of the Pt-Fe bimetallic sites in the cluster-derived samples were altered by exposure to the reactants. The Pt(5)Fe(2)/SiO(2) sample was also more active than Pt/SiO(2) for PROX with a selectivity of approximately 92% at 50 degrees C. In this case, the deactivation with time on stream was substantially slower, indicating that the highly reducing environment under the PROX conditions helps maintain the properties of the active Pt-Fe bimetallic sites. PMID- 16700609 TI - Shell cross-linked Au nanoparticles. AB - The organic layer of thiol-protected Au nanoparticles (ca.3 nm in diameter) was cross-linked using ring-opening metathesis polymerization or Michael addition of polyfunctional amines. The shell cross-linked nanoparticles showed increased stability toward thermal treatment and oxidative etching. The Au core of cross linked nanoparticles was removed in an attempt to prepare hollow capsules. However, Au etching resulted in insoluble materials. PMID- 16700610 TI - Carbon nanotubes with small and tunable diameters from poly(ferrocenylsilane) block-polysiloxane diblock copolymers. AB - Iron-containing nanostructures produced from various self-assembled poly(ferrocenylsilane)-block-polysiloxane thin films are catalytically active for the initiation and growth of high density, small diameter carbon nanotubes (CNTs). Moreover, the tube diameter and density can be tuned by adjusting the chain lengths of the block copolymer. Iron-containing nanostructures from poly(ferrocenylmethylethylsilane)-b-poly(methylvinylsiloxane) polymer with 25 repeat units of an iron-containing segment and 265 repeat units of a non-iron containing segment are able to produce CNTs with diameters around or less than 1 nm. Lithographically selective growth of CNTs across a large surface area has been demonstrated using this polymer system. Under the same growth condition, it has been found that the yield of defect-free CNTs varies with the size of the catalytically active nanostructures, which are dictated by the chain lengths of the two blocks. This result indicates that, for a specific-sized catalyst nanocluster, a unique set of growth conditions is required for synthesizing high yield, defect-free CNTs. This finding further addresses the importance of using uniform-sized catalyst-containing nanostructures for consistently achieving high yield and high-quality CNTs with a minimum number of defects and amount of amorphous carbon. PMID- 16700611 TI - Simulation of nonwetting phase entrapment within porous media using magnetic resonance imaging. AB - Models representing the pore structures of amorphous, mesoporous silica pellets have been constructed using magnetic resonance images of the materials. Using magnetic resonance imaging (MRI), maps of the macroscopic (approximately 0.01-1 mm) spatial distribution of porosity and pore size were obtained. The nature and key parameters of the physical mechanism for mercury retraction, during porosimetry experiments on the silica materials, were determined using integrated gas sorption experiments. Subsequent simulations of mercury porosimetry within the structural models derived from MRI have been used to successfully predict, a priori, the point of the onset of structural hysteresis and the final levels of mercury entrapment for the silicas. Hence, a firm understanding of the physical processes of mercury retraction and entrapment in these amorphous silica materials has been established. PMID- 16700612 TI - In situ scanning tunneling microscopy of 1,6-hexanedithiol, 1,9-nonanedithiol, 1,2-benzenedithiol, and 1,3-benzenedithiol adsorbed on pt(111) electrodes. AB - Cyclic voltammetry (CV) and in situ scanning tunneling microscopy (STM) were used to examine four dithiol molecules, including 1,6-hexanedithiol, 1,9 nonanedithiol, 1,2-benzenedithiol, and 1,3-benzenedithiol, adsorbed on well ordered Pt(111) electrodes in 0.1 M HClO(4). The open-circuit potential (OCP) of Pt(111) electrodes decreased substantially from 0.95 to 0.3 V (versus reversible hydrogen electrode) upon the adsorption of dithol molecules, which indicates that these adsorbates injected electrons into the Pt electrode. For all dithiol molecules, ordered adlattices of p(2 x 2) and (square root 3 x square root 3)R30 degrees were formed when the dosing concentration was lower than 150 microM and the potential of Pt(111) was more negative than 0.5 V. Raising the potential of Pt(111) from 0.1 to 0.4 V or more positive values could transform p(2 x 2) to (square root 3 x square root 3)R30 degrees before it turned disarray. The insensitivity of the structure of dithiol adlayers with their chemical structures was explained by upright molecular orientation with the formation of one Pt-S bond per dithiol molecule. This molecular orientation was independent of the coverage of dithiol molecules, as nucleation seeds produced at the beginning of adsorption were also constructed with p(2 x 2). The triangular-shaped STM molecular resolution suggested 3-fold binding of sulfur headgroup on Pt(111). All dithiols were adsorbed so strongly on Pt(111) electrodes that switching the potential negatively to the onset of hydrogen evolution in 0.1 M HClO(4) or water reduction in 1 M KOH could not displace dithiol admolecules. PMID- 16700613 TI - Spectroelectrochemistry and preresonance Raman spectroscopy of polyaniline dodecylbenzenesulfonic acid colloidal dispersions. AB - Dodecylbenzenesulfonic acid (DBSA) dissolved in 2-propanol (70% solution), a surfactant and dopant, is used in the synthesis of polyaniline (PANI) colloids. The micelle-aided synthesis yields green dispersions that do not undergo macroscopic precipitation for more than a year. UV-vis studies reveal that polymerization is completed in 3 h irrespective of the feed ratio of DBSA/aniline. The polymerization yield depends on the amount of DBSA in the feed. UV-vis spectroscopy as a function of applied potential was carried out for the first time for thin layers of the dispersion. Bands corresponding to both emeraldine salt and emeraldine base were observed during cathodic potential sweeps whereas in an anodic sweep bands due to only emeraldine base were observed. Electronic absorption spectroscopy and preresonance Raman spectroscopy as a function of pH indicate emeraldine salt (metal) to emeraldine base (insulator) transformation at higher pH (pH 9-10). The high aggregation tendency of PANI particles during solvent drying yields particles with irregular shapes. PMID- 16700614 TI - Future prospects for the cure of brain cancer. PMID- 16700615 TI - Molecular biology of human gliomas. AB - Human gliomas are the most common primary central nervous system neoplasm, and they are a complex, heterogeneous, and difficult disease to treat. In the past two decades, advances in molecular biology have revolutionized our understanding of the mechanism by which these neoplasms are initiated and progress. While surgery, radiation therapy, and chemotherapy have roles to play in the treatment of patients with gliomas; these therapies are self-limited because of the intrinsic resistance of glioma cells to therapy, and the diffusely infiltrating nature of the lesions. It is now known that malignant gliomas arise from a number of well-characterized genetic alterations and activations of oncogenes and inactivation of tumor suppressor genes. These genetic alterations disrupt critical cell cycle, growth factor activation, apoptotic, cell motility, and invasion pathways that lead to phenotypic changes and neoplastic transformation. Research in each of these fields has uncovered potential therapeutic targets that look promising for disease control. Gliomas can now be modeled with fidelity and reproducibility using several transgenic and knockout strategies. Transgenic mouse models are facilitating the testing of various therapeutic strategies in vivo. Finally, the recognition of the putative brain tumor stem cell, the tumor initiating cell in brain cancer, provides an enticing target through which we could eliminate the source of the brain tumor with increased efficacy and less toxicity to normal tissues. In this review, we provide an up-to-date discussion of the many of key technologies and tools that are being used in molecular biology to advance our understanding of the biological behavior of human malignant gliomas. PMID- 16700616 TI - Overexpression of IGFBP5, but not IGFBP3, correlates with the histologic grade of human diffuse glioma: a tissue microarray and immunohistochemical study. AB - Insulin-like growth factor binding proteins (IGFBPs) comprise a family of proteins that bind and regulate the functions of IGFs. One of the IGFBPs, IGFBP2, has recently been shown to be overexpressed in glioblastoma. Overexpression of IGFBP2 contributes to the invasiveness of glioblastoma and correlates with histologic grade and survival in patients with diffuse gliomas, suggesting that IGFBP2 expression may contribute to the glioma formation and/or progression. The expression of other IGFBP family members in gliomas has, however, not been examined in detail. This study was designed to evaluate the expression of IGFBP3 and IGFBP5 in diffuse gliomas using immunohistochemistry applied to a tissue array constructed from 259 gliomas, including ten gliosarcomas. Expression of IGFBP5 correlated significantly with glioma histologic grade. 83% (58/70) of glioblastomas (WHO Grade IV) were positive for IGFBP5, which was significantly higher than WHO Grade III gliomas (41%, 41/101) or WHO Grade II gliomas (18%, 13/72) (p<0.001). In contrast, IGFBP3 was expressed in 17%, 23%, and 17% of WHO Grade IV, Grade III, and Grade II gliomas, respectively (p>0.05). There was no significant difference in IGFBP3 expression among different grades of glioma. Our study thus demonstrates that the expression of IGFBP5, but not IGFBP3, increases with glioma anaplastic progression. The strong correlation between overexpression of IGFBP5 and histologic grade suggests that, in addition to IGFBP2, IGFBP5 may also play a role in glioma progression. PMID- 16700617 TI - Convection-enhanced and local delivery of targeted cytotoxins in the treatment of malignant gliomas. AB - Despite advances in our knowledge about the genesis, molecular biology, and natural history of malignant gliomas and the use of a multi-disciplinary approach to their treatment, patients harboring this diagnosis continue to face a grim prognosis. At the time of diagnosis, patients typically undergo surgery for the establishment of a histologic diagnosis, the reduction of tumor burden, and the relief of mass effect, with the maintenance of the patient's neurological function in mind. This is followed by the administration of adjuvant therapeutics, including radiation therapy and chemotherapy. Many investigational agents with laboratory evidence of efficacy against malignant gliomas have not met their promise in the clinical setting, largely due to the barriers that they must overcome to reach the tumor at a therapeutically meaningful concentration for a durable period of time. The relevant aspects of the blood-brain barrier, blood-tumor barrier, and blood-cerebrospinal fluid barrier, as they pertain to the delivery of agents to the tumor, will be discussed along with the strategies devised to circumvent them. This discussion will be followed by a description of agents currently in preclinical and clinical development, many of which are the result of intense ongoing research into the molecular biology of gliomas. PMID- 16700618 TI - Molecular diagnostics as a tool to personalize treatment in adult glioma patients. AB - Gliomas, the most frequent primary brain tumors in humans, form a heterogeneous group, encompassing many different histological types and malignancy grades. Within this group, the diffuse infiltrative gliomas are by far the most common in adults. The major representatives in this subgroup are the diffuse astrocytic, oligodendroglial, and mixed oligo-astrocytic tumors. Especially in these diffuse gliomas, the role of molecular diagnostics is rapidly increasing. After summarizing the most relevant genetic aberrations and pathways in these tumors detected up till now, this review will discuss the clinical relevance of this information. Several molecular markers have been identified in diffuse gliomas that carry diagnostic and prognostic information. In addition, some of these and other markers predict the response of these gliomas to particular (chemo)therapeutic approaches. The techniques used to obtain this molecular information, as well as the advantages and disadvantages of the different techniques will be discussed. Finally, future perspectives will be presented with regard to the contribution of molecular diagnostics to tailor-made therapy in glioma patients. PMID- 16700619 TI - Neuro-oncological applications of optical spectroscopy. AB - Advances in optics and molecular imaging have occurred rapidly in the past decade. One technique poised to take advantage of these developments is optical spectroscopy (OS). All optical spectroscopic techniques have in common tissue interrogation with light sources ranging from the ultraviolet (UV) to the infrared (IR) ranges of the spectrum, and collection of information on light reflected (reflectance spectroscopy) or light interactions with tissue and emergence at different wavelengths (fluorescence and Raman spectroscopy). OS can provide information regarding intrinsic tissue optical properties such as tissue structure, nuclear density, and the presence or absence of endogenous or exogenous fluorophores. Among other applications, this information has been used to distinguish tumor from normal brain tissues, to detect tumor margins in intrinsic, infiltrating gliomas, to identify radiation damage to tissues, and to assess tissue viability and predict the onset of apoptosis in vitro and in vivo. Potential applications of OS include detection of specific central nervous system (CNS) structures, such as brain nuclei, identification of cell types by the presence of specific neurotransmitters, and the detection of optically labeled cells or drugs during therapeutic interventions. All have potential utility in neuro-oncology, have been investigated in our laboratories, and will be the subject of this review. PMID- 16700620 TI - Convection-enhanced delivery of interleukin-13 receptor-directed cytotoxin for malignant glioma therapy. AB - The treatment of patients with malignant brain tumors, in particular glioblastoma multiforme (GBM) is very challenging because of their diffuse infiltrative nature and the cytological heterogeneity. The median survival of patients with newly diagnosed GBM is only 12-15 months, and only 8-12% of them survive for two years. Novel approaches for brain tumor therapy are needed. Recently, targeted therapies have emerged as promising modality for cancer targeting. We have discovered that high affinity plasma membrane receptor for interleukin-13 (IL-13), an immune regulatory cytokine, is over-expressed in 60-80% of malignant brain tumors. To target these IL-13R, we generated a chimeric fusion protein, composed of human IL 13 and mutated Pseudomonas exotoxin (PE), termed IL-13 cytotoxin (IL13-PE), and tested its cytotoxicity to IL-13R-expressing GBM cells. IL-13 cytotoxin was highly potent and selective in killing IL-13R-expressing GBM cells. In contrast, normal cells including brain, immune, and endothelial cells were generally not affected by this cytotoxin due to no or low expression of IL-13R. In vivo pre clinical studies for safety and toxicity were also performed in mice, rats, and monkeys, and IL-13 cytotoxin was found to be well tolerated by both systemic and intracerebral administrations. IL-13 cytotoxin was found to mediate remarkable efficacy in animal models of human brain tumors. Encouraged by these pre-clinical studies, four Phase 1/2 clinical trials in adult patients with recurrent malignant glioma have been completed. These clinical trials involved convection enhanced delivery (CED) of IL-13 cytotoxin either intratumoral or intraparenchymal after resection of tumor. CED is a novel loco-regional drug delivery method for intracranial tumors that relies on a continuous pressure gradient to distribute drug into interstitial space. This route of IL-13 cytotoxin administration appears to be very well tolerated and have a good risk benefit profile. Most recently, a randomized controlled Phase 3 clinical trial (PRECISE) with intraparenchymal IL-13 cytotoxin administration was completed and subjects are being monitored for safety and survival. PMID- 16700621 TI - Targeting stem cells in brain tumors. AB - Stem cells are commonly defined as undifferentiated cells capable of self renewing and giving rise to a large number of differentiated progeny. It is becoming increasingly apparent that there exist cancer stem cells (CSCs) from which the cells of any given malignancy arise, whereby only a few cells out of a population of cancer cells are able to initiate tumor formation. These CSCs, like their normal counterparts, are characterized by self-renewal and the ability to "differentiate" into all of the cell types in the original tumor. Current chemotherapeutic strategies involve using non-specific cytotoxic agents that target rapidly cycling cells. Although this may reduce disease burden in many cases, these therapies may miss the rare, self-renewing population that truly gives rise to the malignancy (the CSC). This review will focus on the recent discovery of stem cell-like cells in human brain tumors, putative "brain cancer stem cells," which exhibit the properties of self-renewal and the ability to recapitulate the original tumor heterogeneity. Dissecting the molecular mechanisms that underlie the ability of these cells to self-renew and maintain quiescence may allow the development of novel therapeutic strategies that will allow for more efficacious and less toxic therapies for these devastating malignancies. PMID- 16700622 TI - RNAi based approaches to the treatment of malignant glioma. AB - RNA interference (RNAi) is a recently discovered, powerful molecular mechanism that can be harnessed to engineer gene-specific silencing in mammalian tissues. A mechanism, where short double-stranded RNA (dsRNA) molecules, when introduced into cells elicit specific "knock-down" of gene expression via degradation of targeted messenger RNA, has lately become the technique of choice for analysis of gene function in oncology research. Thus, RNAi is currently being extensively evaluated as a potential therapeutic strategy against malignant gliomas, since surgical, radiological, and chemotherapeutic interventions during the past few decades have done little to improve the poor prognosis rate for patients with these dreaded tumors. This review summarizes the pre-clinical studies that are currently underway to test the validity of RNAi as a potential therapeutic strategy against malignant gliomas, and discusses the potential technical hurdles that remain to be overcome before the technique can become a promising clinical therapy to combat this frequently lethal disease. PMID- 16700623 TI - Downregulation of PIK3CB by siRNA suppresses malignant glioma cell growth in vitro and in vivo. AB - EGFR overexpression is the most frequent and important molecular event in the development of astrocytic gliomas, and the P13K signaling pathway is one of the most important downstream pathways of EGFR. EGFR and other members of the receptor tyrosine kinases (RTKs) family, such as VEGFR, PDGFR, and IGFR, et cetera, are often overexpressed in most of malignant gliomas and share common downstream signaling pathways. Therefore, it is considered that directly targeting the downstream PI3K pathway may be more effective in blocking multiple inputs. The PIK3CB gene encoding the class 1A PI3K catalytic subunit p110beta was selected as the target of therapeutic approach for malignant gliomas in the present study. Human U251 glioblastoma cells with high endogenous p110beta expression were transfected with plasmid-based siRNA targeting PIK3CB gene. It was found that downregulation of p110beta expression resulted in the suppression of cell proliferation, arrest of cell cycle, reduction of cell invasion, and promotion of cell apoptosis in vitro. In addition, the growth of the subcutaneous U251 glioma in the nude mice treated with siRNA targeting PIK3CB was significantly inhibited. These results demonstrate that PIK3CB overexpression may play an oncogenic role in the PI3K pathway, and the plasmid-based siRNA targeting of PIK3CB is a potential and promising approach for the treatment of malignant gliomas. PMID- 16700624 TI - Locoregional proton radiotherapy of a primary cavernous sinus non-Hodgkin's lymphoma: case report. AB - Primary B cell lymphomas of the skull base are uncommon and lack well-defined treatment guidelines. We report a case of diffuse, large B-cell lymphoma of the cavernous sinus with sphenoid sinus and clivial extension, treated with partial resection, chemotherapy, and proton beam irradiation. To our knowledge, this is the first report of a skull-base lymphoma treated with protons. A 53-year-old female presented with a two-month history of diplopia, persistent headaches, and paresthesia over the left side of her mouth. A skull MRI revealed an enhancing mass in the right cavernous sinus and right sphenoid sinus. Transsphenoidal subtotal resection of the mass confirmed the presence of a diffuse, large B-cell lymphoma. Treatment consisted of CHOP-R chemotherapy and locoregional radiation with protons. Locoregional radiation of the lesion required moderate doses, below the radiation tolerance of adjacent normal structures. Conformal protons were utilized to minimize the volume of normal brain receiving radiation. Conformal proton beam radiotherapy to a moderate dose proved valuable in this case because it minimized the volume of normal brain receiving low to moderate doses of radiation. PMID- 16700625 TI - Multi-detector CT in the evaluation of patients with recurrence of rectal cancer. AB - The advantages of multiplanar reconstruction in rectal cancer recurrence diagnostics using medium resolution multi-detector CT are evaluated. We included 40 patients after a rectal cancer operation in this study. During follow-up ten patients developed a recurrence. All patients received a minimum of two CT examinations in their follow-up program. A total of 131 CT-scans were evaluated. Each examination was reviewed by two experienced radiologists in respect to recurrence. Each examination was presented in axial reconstruction with a slice thickness of 8mm with an increment of 7 mm and a slice thickness of 3 mm with an increment of 2 mm. The thin slices were used for the multi-planar reconstruction. Multi-planar reconstructions showed better results for the detection of recurrence than axial reconstruction. A reduced slice thickness did not lead to better results in axial reconstruction. Multi-planar reconstruction showed a sensitivity of 0.88, a specificity of 1.0 and an accuracy of 0.97. Our axial reconstruction results were: 0.86, 0.96, and 0.93, respectively. Sensitivity and accuracy showed a significant increase after the first and second examinations. Multi-planar reconstructions allow for better detection of rectal cancer recurrence when compared to axial reconstructions. Thinner axial slice thickness shows no diagnostic advantage. PMID- 16700626 TI - Identification of primary tumors of brain metastases by infrared spectroscopic imaging and linear discriminant analysis. AB - This study applies infrared (IR) spectroscopy to distinguish normal brain tissue from brain metastases and to determine the primary tumor of four frequent brain metastases such as lung cancer, colorectal cancer, breast cancer, and renal cell carcinoma. Standard methods sometimes fail to identify the origin of brain metastases. As metastatic cells contain the molecular information of the primary tissue cells and IR spectroscopy probes the molecular fingerprint of cells, IR spectroscopy based methods constitute a new approach to determine the primary tumor of a brain metastasis. IR spectroscopic images were recorded by a FTIR spectrometer equipped with a macro sample chamber and coupled to a focal plane array detector. Unsupervised cluster analysis of IR images revealed variances within each sample and between samples of the same tissue type. Cluster averaged IR spectra of tissue classes with known diagnoses were selected to develop a metric with eight variables. These data trained a supervised classification model based on linear discriminant analysis that was used to identify the origin of 20 cryosections including one brain metastasis with an unknown primary tumor. PMID- 16700627 TI - Interacting adaptive processes with different timescales underlie short-term motor learning. AB - Multiple processes may contribute to motor skill acquisition, but it is thought that many of these processes require sleep or the passage of long periods of time ranging from several hours to many days or weeks. Here we demonstrate that within a timescale of minutes, two distinct fast-acting processes drive motor adaptation. One process responds weakly to error but retains information well, whereas the other responds strongly but has poor retention. This two-state learning system makes the surprising prediction of spontaneous recovery (or adaptation rebound) if error feedback is clamped at zero following an adaptation extinction training episode. We used a novel paradigm to experimentally confirm this prediction in human motor learning of reaching, and we show that the interaction between the learning processes in this simple two-state system provides a unifying explanation for several different, apparently unrelated, phenomena in motor adaptation including savings, anterograde interference, spontaneous recovery, and rapid unlearning. Our results suggest that motor adaptation depends on at least two distinct neural systems that have different sensitivity to error and retain information at different rates. PMID- 16700628 TI - High guanine and cytosine content increases mRNA levels in mammalian cells. AB - Mammalian genes are highly heterogeneous with respect to their nucleotide composition, but the functional consequences of this heterogeneity are not clear. In the previous studies, weak positive or negative correlations have been found between the silent-site guanine and cytosine (GC) content and expression of mammalian genes. However, previous studies disregarded differences in the genomic context of genes, which could potentially obscure any correlation between GC content and expression. In the present work, we directly compared the expression of GC-rich and GC-poor genes placed in the context of identical promoters and UTR sequences. We performed transient and stable transfections of mammalian cells with GC-rich and GC-poor versions of Hsp70, green fluorescent protein, and IL2 genes. The GC-rich genes were expressed several-fold to over a 100-fold more efficiently than their GC-poor counterparts. This effect was not due to different translation rates of GC-rich and GC-poor mRNA. On the contrary, the efficient expression of GC-rich genes resulted from their increased steady-state mRNA levels. mRNA degradation rates were not correlated with GC content, suggesting that efficient transcription or mRNA processing is responsible for the high expression of GC-rich genes. We conclude that silent-site GC content correlates with gene expression efficiency in mammalian cells. PMID- 16700629 TI - Fgf9 and Wnt4 act as antagonistic signals to regulate mammalian sex determination. AB - The genes encoding members of the wingless-related MMTV integration site (WNT) and fibroblast growth factor (FGF) families coordinate growth, morphogenesis, and differentiation in many fields of cells during development. In the mouse, Fgf9 and Wnt4 are expressed in gonads of both sexes prior to sex determination. Loss of Fgf9 leads to XY sex reversal, whereas loss of Wnt4 results in partial testis development in XX gonads. However, the relationship between these signals and the male sex-determining gene, Sry, was unknown. We show through gain- and loss-of function experiments that fibroblast growth factor 9 (FGF9) and WNT4 act as opposing signals to regulate sex determination. In the mouse XY gonad, Sry normally initiates a feed-forward loop between Sox9 and Fgf9, which up-regulates Fgf9 and represses Wnt4 to establish the testis pathway. Surprisingly, loss of Wnt4 in XX gonads is sufficient to up-regulate Fgf9 and Sox9 in the absence of Sry. These data suggest that the fate of the gonad is controlled by antagonism between Fgf9 and Wnt4. The role of the male sex-determining switch--Sry in the case of mammals--is to tip the balance between these underlying patterning signals. In principle, sex determination in other vertebrates may operate through any switch that introduces an imbalance between these two signaling pathways. PMID- 16700632 TI - Quail poisoning in a child. AB - CONTEXT: It has been well known since ancient times that the consumption of quail may be deleterious to health. The European quail sub-species (Coturnix coturnix coturnix), common to Europe, North Africa, and western Asia is indeed potentially poisonous to humans, but only during the autumn migration (not during their return flight in spring). Quail poisoning is an acute dietary-toxicological syndrome. ISSUE: In this case report, we describe a rare, non-fatal case of quail poisoning in a 12-year-old boy. The victim was admitted to hospital 5 hours after the consumption of quail because of the signs and symptoms, as well as laboratory findings (elevation of serum muscle enzymes, myoglobinuria) significant of acute rhabdomyolysis. He was treated symptomatically with bed rest, forced continuous diuresis, urine alkalization, and maintenance of fluid and electrolyte balance. The victim recovered completely without sequelae after 8 days hospitalization. LESSONS: The aetiology of this syndrome is still unclear. It has been suggested that a toxic effect or a genetic sensitivity plays a major role. Many cases of quail poisoning in adults have been reported previously on the island of Lesbos during autumn. To our knowledge, this is the first case reported in a child. PMID- 16700631 TI - Genome microevolution of chikungunya viruses causing the Indian Ocean outbreak. AB - BACKGROUND: A chikungunya virus outbreak of unprecedented magnitude is currently ongoing in Indian Ocean territories. In Reunion Island, this alphavirus has already infected about one-third of the human population. The main clinical symptom of the disease is a painful and invalidating poly-arthralgia. Besides the arthralgic form, 123 patients with a confirmed chikungunya infection have developed severe clinical signs, i.e., neurological signs or fulminant hepatitis. METHODS AND FINDINGS: We report the nearly complete genome sequence of six selected viral isolates (isolated from five sera and one cerebrospinal fluid), along with partial sequences of glycoprotein E1 from a total of 127 patients from Reunion, Seychelles, Mauritius, Madagascar, and Mayotte islands. Our results indicate that the outbreak was initiated by a strain related to East-African isolates, from which viral variants have evolved following a traceable microevolution history. Unique molecular features of the outbreak isolates were identified. Notably, in the region coding for the non-structural proteins, ten amino acid changes were found, four of which were located in alphavirus-conserved positions of nsP2 (which contains helicase, protease, and RNA triphosphatase activities) and of the polymerase nsP4. The sole isolate obtained from the cerebrospinal fluid showed unique changes in nsP1 (T301I), nsP2 (Y642N), and nsP3 (E460 deletion), not obtained from isolates from sera. In the structural proteins region, two noteworthy changes (A226V and D284E) were observed in the membrane fusion glycoprotein E1. Homology 3D modelling allowed mapping of these two changes to regions that are important for membrane fusion and virion assembly. Change E1-A226V was absent in the initial strains but was observed in >90% of subsequent viral sequences from Reunion, denoting evolutionary success possibly due to adaptation to the mosquito vector. CONCLUSIONS: The unique molecular features of the analyzed Indian Ocean isolates of chikungunya virus demonstrate their high evolutionary potential and suggest possible clues for understanding the atypical magnitude and virulence of this outbreak. PMID- 16700633 TI - Predicting negative mood state and personal growth in African American and White long-term breast cancer survivors. AB - BACKGROUND: Relatively little research has examined cognitive processes that may impact psychological adaptation in older long-term breast cancer survivors (BCS). PURPOSE: This study investigated the strength of a conceptual model based on the literature and Uncertainty in Illness Theories which proposes that negative mood state and personal growth in older long-term White and African American BCS would be predicted by the combined influences of demographic and disease variables, social support, religious participation, and cognitive processes (uncertainty, catastrophizing, troublesome thoughts, and cognitive reframing). METHODS: Baseline data were gathered from 524 BCS (369 Whites and 155 African Americans, 5 9 years postdiagnosis) prior to their participating in an uncertainty management intervention program. The conceptual model was tested using structural equation modeling. RESULTS: The multigroup model showed good fit to the data and explained substantial variance in negative mood state and personal growth. Cognitive processes showed both direct and indirect effects on outcomes in the expected directions. Several ethnic differences were found: African Americans were more negatively affected by comorbidities and Whites by symptom distress, whereas cognitive reframing was a stronger predictor of personal growth for African Americans than Whites. CONCLUSIONS: This is one of the first studies to explore predictors of both negative mood and personal growth in a multiethnic sample of BCS. These findings suggest that cognitive processes play an important role in psychological adaptation to breast cancer survivorship. These processes are amenable to change, suggesting a logical target for intervention with this population. PMID- 16700634 TI - A systematic review of randomized trials on the effectiveness of computer tailored education on physical activity and dietary behaviors. AB - BACKGROUND: Although computer-tailored promotion of dietary change and physical activity has been identified as a promising intervention strategy, there is a need for a more systematic evaluation of the evidence. PURPOSE: This study systematically reviews the scientific literature on computer-tailored physical activity and nutrition education. METHODS: Intervention studies published from 1965 up to September 2004 were identified through a structured search in PubMed, PsycInfo, and Web of Science and an examination of reference lists of relevant publications. Studies were included that applied a pretest-posttest randomized controlled trial design, were aimed at primary prevention among adults, used computer-tailored interventions to change physical activity and dietary behaviors, and were published in English. The search resulted in 30 publications 11 on physical activity behaviors and 26 on nutrition behaviors, some studies investigated multiple behaviors. RESULTS: Three of 11 of the physical activity studies and 20 of 26 of the nutrition studies found significant effects of the tailored interventions. The evidence was most consistent for tailored interventions on fat reduction. CONCLUSIONS: Overall, there seems to be potential for the application of computer tailoring for promoting healthy diets, but more research is needed to test computer-tailored interventions against other state-of the-art intervention techniques and to identify the mechanisms underlying successful computer tailoring. PMID- 16700630 TI - Lentiviral expression of retinal guanylate cyclase-1 (RetGC1) restores vision in an avian model of childhood blindness. AB - BACKGROUND: Leber congenital amaurosis (LCA) is a genetically heterogeneous group of retinal diseases that cause congenital blindness in infants and children. Mutations in the GUCY2D gene that encodes retinal guanylate cyclase-1 (retGC1) were the first to be linked to this disease group (LCA type 1 [LCA1]) and account for 10%-20% of LCA cases. These mutations disrupt synthesis of cGMP in photoreceptor cells, a key second messenger required for function of these cells. The GUCY1*B chicken, which carries a null mutation in the retGC1 gene, is blind at hatching and serves as an animal model for the study of LCA1 pathology and potential treatments in humans. METHODS AND FINDINGS: A lentivirus-based gene transfer vector carrying the GUCY2D gene was developed and injected into early stage GUCY1*B embryos to determine if photoreceptor function and sight could be restored to these animals. Like human LCA1, the avian disease shows early-onset blindness, but there is a window of opportunity for intervention. In both diseases there is a period of photoreceptor cell dysfunction that precedes retinal degeneration. Of seven treated animals, six exhibited sight as evidenced by robust optokinetic and volitional visual behaviors. Electroretinographic responses, absent in untreated animals, were partially restored in treated animals. Morphological analyses indicated there was slowing of the retinal degeneration. CONCLUSIONS: Blindness associated with loss of function of retGC1 in the GUCY1*B avian model of LCA1 can be reversed using viral vector-mediated gene transfer. Furthermore, this reversal can be achieved by restoring function to a relatively low percentage of retinal photoreceptors. These results represent a first step toward development of gene therapies for one of the more common forms of childhood blindness. PMID- 16700635 TI - Knowledge of heart disease risk in a multicultural community sample of people with diabetes. AB - BACKGROUND: Prevention of coronary heart disease (CHD) is a primary goal of diabetes management. Unfortunately, CHD risk knowledge is poor among people with diabetes. PURPOSE: The objective is to determine predictors of CHD risk knowledge in a community sample of people with diabetes. METHODS: A total of 678 people with diabetes completed the Heart Disease Facts Questionnaire (HDFQ), a valid and reliable measure of knowledge about the relationship between diabetes and heart disease. RESULTS: In regression analysis with demographics predicting HDFQ scores, sex, annual income, education, and health insurance status predicted HDFQ scores. In a separate regression analysis, having CHD risk factors did not predict HDFQ scores, however, taking medication for CHD risk factors did predict higher HDFQ scores. An analysis of variance showed significant differences between ethnic groups for HDFQ scores; Whites (M = 20.9) showed more CHD risk knowledge than African Americans (M = 19.6), who in turn showed more than Latinos (M = 18.2). Asians scored near Whites (M = 20.4) but did not differ significantly from any other group. Controlling for numerous demographic, socioeconomic, health care, diabetes, and cardiovascular health variables, the magnitude of ethnic differences was attenuated, but persisted. CONCLUSION: Education regarding modifiable risk factors must be delivered in a timely fashion so that lifestyle modification can be implemented and evaluated before pharmacotherapy is deemed necessary. African Americans and Latinos with diabetes are in the greatest need of education regarding CHD risk. PMID- 16700636 TI - Neuropsychological characteristics of adolescent boys differing in risk for high blood pressure. AB - BACKGROUND: Individuals with established hypertension have been found to display deficits in a number of neuropsychological abilities. In general, these are probably due to structural changes in the brain produced by sustained high blood pressure. However, a potentially important line of research suggests that some of these deficits may extend to younger individuals with less severe elevations of blood pressure, perhaps even children, and thus be related more to risk for hypertension than hypertension per se. PURPOSE: The objective was to examine the relationships between neuropsychological function and risk for hypertension in children. METHODS: Measurements of blood pressure and parental history of hypertension were obtained in 88 French-Canadian 14-year-old boys and used to predict performance on a neuropsychological battery. RESULTS: Boys at greater risk of hypertension by virtue of having a parental history of high blood pressure and normatively elevated systolic blood pressure had significantly lower scores on a verbal learning factor score compared to boys at lower risk. Boys with normatively elevated systolic blood pressure also had significantly lower scores on a spatial learning and memory factor score compared to boys with lower blood pressure. The results could not be attributed to differences in family socioeconomic status. CONCLUSIONS: Using a younger sample than typically employed in the area, the results support previous suggestions that some of the neuropsychological characteristics displayed by hypertensive individuals may predate the development of clinically elevated blood pressure and could be associated with risk for the disorder. PMID- 16700638 TI - Blunted cardiovascular responses to daytime activities as related to reduced nocturnal blood pressure decline. AB - BACKGROUND: Individuals showing less than a 10% decline in blood pressure at night ("nondippers") are known to be at increased risk for hypertension and other cardiovascular conditions. PURPOSE: This research tested the assertion by Raikkonen et al. (1) that nondippers show blunted cardiovascular responses to activities during daytime hours. METHODS: Ambulatory blood pressure and impedance monitoring was performed with 149 young adults in Singapore. At each daytime blood pressure reading, participants completed a computerized questionnaire indicating location, posture, physical activities, feelings, and social interactions. RESULTS: Significant interactions between dipper status and feeling tired, location, posture, and physical activity provided qualified support for the blunting hypothesis. However, blunting of BP responses was not attributable to blunting of either cardiac output or total peripheral resistance responses. CONCLUSION: Nondippers appear to show blunted daytime responses to certain types of daytime activities. However, these effects are limited and appear to be the result of different mechanisms than those responsible for reduced nighttime blood pressure decline. PMID- 16700637 TI - Paradoxical effects of social support on blood pressure reactivity among defensive individuals. AB - BACKGROUND: Social support can reduce cardiovascular responses to an acute stressor. However, prior clinical research suggests that defensive individuals may react negatively to social support. PURPOSE: This experiment examined whether emotional support provided during a speech stressor would escalate rather than decrease blood pressure (BP) reactivity among defensive individuals. METHODS: After completing personality measures, 176 female undergraduates were randomly assigned to give a speech in 1 of 3 social conditions: alone, or with a neutral or supportive confederate present. Mean arterial BP was assessed at baseline, immediately before, and during and after the stressor. RESULTS: In the supportive condition, defensiveness predicted higher BP reactivity during anticipation (beta = .35, p = .04) and delivery of the speech (beta = .32, p = .03), and longer recovery to baseline BP levels (beta = .35, p = .01). In the absence of support (i.e., alone and neutral conditions), defensiveness was not associated with BP reactivity. Defensiveness also predicted greater dissociation between subjective stress and BP responses (beta = .35, p = .05), but only in the supportive condition. CONCLUSIONS: According to results, the provision of social support during stressors does not benefit defensive individuals' BP reactivity but has the opposite effect. PMID- 16700639 TI - The relation of free plasma tryptophan to anger, hostility, and aggression in a nonpatient sample of adult men and women. AB - BACKGROUND: Dysregulation of central nervous system serotonergic (5-HT) activity is implicated in behavioral states and psychological traits associated with depression and aggression, with some studies suggesting possible gender-related differences. PURPOSE: This study examined the relation of free plasma tryptophan (TRP) to aggression and depression in a sample of 138 nonsmoking adults recruited from the general community. It was hypothesized that TRP would be associated with anger, hostility, and aggression. METHODS: To minimize effects of diurnal variation and menstrual cycle, fasting blood samples were collected in the morning, and, for women, during the follicular phase of the menstrual cycle. Participants were administered questionnaires following blood draw. Plasma TRP was determined by high performance liquid chromatography. RESULTS: In women, but not men, higher levels of TRP were associated with trait hostility, propensity for anger, a tendency to express anger outwardly, and an antagonistic interpersonal style. For men and women, greater severity of depressive symptoms, anger, and the verbal expression of anger were associated with higher TRP. These associations were independent of age, body mass index, fasting albumin, and race and ethnicity. CONCLUSIONS: These data suggest that in women, but not men, higher plasma levels of TRP, the precursor to 5-HT, are associated with anger-hostility aggression and that these associations are independent of various potential confounds. Implications of these observations to studies employing acute TRP depletion studies are discussed. PMID- 16700640 TI - A randomized clinical trial of group-based cognitive-behavioral stress management in localized prostate cancer: development of stress management skills improves quality of life and benefit finding. AB - BACKGROUND: Recent literature has indicated that a significant percentage of oncology patients describe finding some benefit (e.g., improved personal growth, sense of meaning, and enhanced interpersonal relationships) in the cancer experience. However, few studies have investigated the role of group-based psychosocial interventions in improving benefit finding (BF), and virtually none have investigated these constructs in men. PURPOSE: This study examined whether a cognitive-behavioral stress management (CBSM) intervention improves BF and quality of life (QoL) in men recovering from treatment for localized prostate cancer. METHODS: Participants in this study were 191 men (M age = 65.1) treated with radiation or radical prostatectomy for clinically localized (i.e., Stage I or II) prostate cancer. Participants were primarily non-Hispanic White (40%) or Hispanic (41%), followed by Black (18%) and other ethnicity (1%), were an average of 65.1 years old (SD = 7.7), and earned an average of 47,800 US dollars annually (SD = 41,000 US dollars). Participants were randomized to either a 10-week group based cognitive-behavioral stress management intervention or a half-day educational seminar as a control condition. All participants provided demographic information and completed the Positive Contributions Scale-Cancer to assess BF, the Functional Assessment of Cancer Therapy to measure quality of life, and a measure of perceived stress management skills. Structural equation modeling was utilized for all analyses. RESULTS: Results indicated that the CBSM condition led to increases in BF and QoL and that these changes were mediated by the development of stress management skills. CONCLUSIONS: Results support the use of group-based cognitive-behavioral interventions in promoting QoL and BF in this population. PMID- 16700641 TI - Hystersisters online: social support and social comparison among hysterectomy patients on the internet. AB - BACKGROUND: The Internet has become a popular source of health information for patients with a variety of medical concerns; however, research examining patient interactions on the Internet has been limited. PURPOSE: Four questions were examined in a survey study of hysterectomy patients who visited http://www.hystersisters.com: (a) Do hysterectomy patients use the support Web site because they perceive their proximal sources of social support to be inadequate? (b) What kinds of support do patients receive from the Web site? (c) What attributes characterize the "Hystersisters" that are perceived to be most helpful? (d) How do informational sources vary depending on the nature of the patient concern? METHODS: Women (N = 137) responded to questions about social support, Web site use, and perceptions of other Web site users. RESULTS: Participants reported high levels of perceived support and tangible assistance from their proximal social environment during recovery from surgery (93%-100%). Hystersisters who were perceived as helpful tended to share similar attributes, such as religion and children. On the Web site, information and advice (61%) was sought significantly more than emotional or esteem support (p < .01). For issues involving spiritual or partner matters versus factual issues connected to the hysterectomy, patients expressed greater interest in communicating with a patient who shared their values even if they were not more knowledgeable. CONCLUSIONS: This research contributes to our understanding of how patients utilize the Internet for health information. Longitudinal research is needed to evaluate causal relationships between Internet use and health outcomes. PMID- 16700642 TI - At-risk drinking in employed men and women. AB - BACKGROUND: "At-risk" drinking is associated with a variety of negative health and social consequences. However, little is known about the characteristics of at risk drinkers or of changes in at-risk status over time. PURPOSE: The objective was to examine the correlates of at-risk drinking and the prospective predictors of maintenance or change in at-risk status. METHOD: Participants were 4,322 employed individuals assessed at baseline and 4 years later. At-risk drinking was defined as 2 or more drinks per day for men and 1 or more drinks per day for women. RESULTS: The baseline prevalence of at-risk drinking was 11%. Four percent of baseline not-at-risk individuals transitioned to at-risk drinking at follow up, and 54% of the baseline at-risk individuals remained at-risk at follow-up. Several demographic-, work-, and tobacco-related variables differentiated at-risk groups and were prospective predictors of change in at-risk drinking status among those individuals who were not at risk at baseline. However, none of the constructs predicted change among at-risk drinkers. CONCLUSION: The data suggest that at-risk drinking is of public health concern. Eleven percent of the participants met criteria for at-risk drinking. Further, at-risk and not-at-risk drinkers differed on numerous characteristics, and their drinking may be influenced by different factors. PMID- 16700643 TI - Physician vaccinate thyself: why influenza vaccination rates are higher among clinicians than among nonclinicians. AB - BACKGROUND: National vaccination guidelines recommend influenza vaccination for clinical employees with patient contact. For many other healthy working adults, influenza vaccination is optional. PURPOSE: We examined acceptance of a free influenza vaccine among clinical university employees who had patient contact and nonclinical university employees who did not have patient contact. We asked whether and why job role had an influence on the decision to be vaccinated. METHOD: Clinical (n = 103) and nonclinical (n = 339) employees of two universities completed a mailed questionnaire. RESULTS: Vaccination rates were higher among clinical employees (62%) than among their nonclinical counterparts (47%). This effect of job role on vaccine acceptance was statistically mediated by perceived riskiness of getting vaccinated, knowledge about the vaccine, easy access to the vaccine, pro-Western medicine beliefs, and social influence. It was not vaccinated by perceived likelihood of getting infected or concern about infecting others. CONCLUSION: Clinical employees have higher vaccination rates than nonclinical employees, but not for the reasons that form the basis of national vaccination guidelines. PMID- 16700644 TI - The course and prognosis of hip complaints in general practice. AB - BACKGROUND: The general practitioner needs to discriminate complaints with need of specialist care from those that can be managed in primary care. However, no previous research has studied prognostic indicators for the course of hip complaints in a primary care population. PURPOSE: The purpose of this study was to investigate the course of hip complaints presented in general practice and to identify relevant prognostic indicators of outcome. METHODS: Data were collected by means of self-administered questionnaires containing questions about sociodemographic variables, characteristics of the complaints, and several intraindividual and extraindividual factors, including several psychosocial variables (e.g., pain coping, distress, and kinesiophobia). After 3 and 12 months of follow-up perceived recovery, change in pain intensity and change in functioning were assessed. Multiple regression analyses were performed to investigate the association between the potential prognostic indicators and the 3 outcome measures. RESULTS: We included 139 patients with hip complaints, presented in general practice. Only 24% reported recovery after 3 months, increasing to 37% after 12 months. A history of hip complaints, a longer duration of the current episode of hip complaints, or more severe complaints, were associated with a less favorable prognosis. Furthermore, more vital patients and patients who met the Norm for Healthy Activity had a higher probability of a favorable outcome. Pain transformation and worrying were significant associated with recovery and changes in functioning after 3 months. CONCLUSIONS: Different prognostic indicators were found to be associated with perceived recovery, changes in pain intensity, and changes in functioning. Future research should aim at investigating the mechanisms that can underlie these associations. PMID- 16700648 TI - Mechanisms of calcium signaling and function in lymphocytes. AB - The multifunctional role for calcium during lymphocyte activation and differentiation is well established; yet how fate-specific changes in calcium concentration are produced, and how these signals are translated into distinct cellular responses, is not known. A critical role has been identified for store operated calcium release-activated calcium (CRAC) channels; however, the lack of information concerning the structure and mechanism of store-dependent activation has limited our ability to define CRAC's role in the range of calcium-dependent responses of lymphocytes. Moreover, additional calcium-permeant cation channels have been recently identified in lymphocytes. These channels regulate functions distinct from those attributed to CRAC channels. Consequently, this review summarizes well-established mechanisms of calcium signaling and also considers recent findings which suggest that diverse and interacting pathways encode functionally specific calcium signals in lymphocytes. Speculation is also offered about how distinct patterns of calcium signaling are translated into diverse transcriptional responses. PMID- 16700649 TI - The role of protein kinase A and A-kinase anchoring proteins in modulating T-cell activation: progress and future directions. AB - Protein kinase A (PKA) is a broad-specificity serine/threonine protein kinase whose spatial and temporal regulation is maintained through interactions with A kinase anchoring proteins (AKAPs). Subcellular localization of AKAPs through unique targeting domains provides a mechanism by which PKA can respond to localized microdomains of cyclic AMP (cAMP) and phosphorylate nearby substrates. For nearly 40 years, cAMP has been known to be a potent modulator of the immune system. cAMP levels are regulated by G-protein-coupled receptors, adenylyl cyclases (AC), and phosphodiesterases (PDEs). This review discusses recent progress made in the discovery of PKA substrates in T lymphocytes and in the identification of AKAPs in T lymphocytes. Because PKA is activated by cAMP, generation and maintenance of cAMP in T cells is also discussed. These findings are framed in the context of understanding the complexity of cAMP and, thus, PKA signaling and are intended to provide the reader with an overview of current literature, as well as an awareness of questions and concerns to consider. PMID- 16700650 TI - Caspase function and the immune system. AB - Caspases are key effector components of apoptosis, the process of programmed cell death. However, a large body of recent work has shown that caspase activity is necessary for several nonapoptotic cellular functions, such as proliferation, differentiation, survival, and the regulation of cytokine expression. In this review, we concentrate on the nonapoptotic functions of caspases and, in particular, on nonapoptotic roles of caspases in the immune system. Because of the involvement of caspases in promoting cell death, as well as cell proliferation and other nonapoptotic functions, studies involving the inhibition of caspases in vivo must be interpreted with caution. PMID- 16700651 TI - Innate and adaptive immune responses of the central nervous system. AB - The central nervous system (CNS) is an immunologically specialized organ. The blood-brain barrier regulates the passage of molecules and cells into the CNS. Robust immune responses occur in the CNS even though there is normally an absence of MHC molecules, lack of normal lymphatic drainage, and reduced immune surveillance. This review discusses the immunological elements of the healthy CNS and the pattern of responses that evolve during innate and adaptive immunity in this organ. We also discuss the contribution of astrocytes, cerebrovascular endothelial cells, microglia, macrophages, and dendritic cells to the integrity and pathology of the CNS during CD4+ T-cell autoimmune responses directed against neuroantigens. PMID- 16700652 TI - Cardiovascular tissue engineering I. Perfusion bioreactors: a review. AB - Tissue engineering is a fast-evolving field of biomedical science and technology with future promise to manufacture living tissues and organs for replacement, repair, and regeneration of diseased organs. Owing to the specific role of hemodynamics in the development, maintenance, and functioning of the cardiovascular system, bioreactors are a fundamental of cardiovascular tissue engineering. The development of perfusion bioreactor technology for cardiovascular tissue engineering is a direct sequence of previous historic successes in extracorporeal circulation techniques. Bioreactors provide a fluidic environment for tissue engineered tissue and organs, and guarantee their viability, maturation, biomonitoring, testing, storage, and transportation. There are different types of bioreactors and they vary greatly in their size, complexity, and functional capabilities. Although progress in design and functional properties of perfusion bioreactors for tissue engineered blood vessels, heart valves, and myocardial patches is obvious, there are some challenges and insufficiently addressed issues, and room for bioreactor design improvement and performance optimization. These challenges include creating a triple perfusion bioreactor for vascularized tubular tissue engineered cardiac construct; designing and manufacturing fluidics-based perfused minibioreactors; incorporation of systematic mathematical modeling and computer simulation based on computational fluid dynamics into the bioreactor designing process; and development of automatic systems of hydrodynamic regime control. Designing and engineering of built-in noninvasive biomonitoring systems is another important challenge. The optimal and most efficient perfusion and conditioning regime, which accelerates tissue maturation of tissue-engineered constructs also remains to be determined. This is a first article in a series of reviews on critical elements of cardiovascular tissue engineering technology describing the current status, unsolved problems, and challenges of bioreactor technology in cardiovascular tissue engineering and outlining future trends and developments. PMID- 16700653 TI - Bone contact around osseointegrated implants: a histologic study of acid-etched and machined surfaces. AB - Current literature reports that surface acid etching can improve bone--implant contact (BIC). The aim of this study was to evaluate the differences of BIC between acid-etched (Osseotite) and machined surface implants. Six white New Zealand mature rabbits were used in the present investigation. Each rabbit received two implants, specially made with two surfaces on it (one acid-etched and one machined) into each tibia. A total of 24 implants were inserted. Two animals each were killed at 15, 30, and 60 days after implant placement. Histomorphometry of BIC was statistically evaluated. The acid-etched surfaces showed a greater bone contact percentage than the machined ones during the early phase of healing (15 days), which was not statistically significant. On the other hand, after 30 and 60 days, the differences of BIC between acid-etched and machined surfaces were statistically significant. The acid-etched surface appears to improve BIC at a later stage of healing. PMID- 16700654 TI - Histological evaluation, in rabbit tibiae, of osseointegration of mini-implants in sites prepared with Er:YAG laser versus sites prepared with traditional burs. AB - OBJECTIVE: The aim of this study was to evaluate the different bone reactions in rabbit tibiae after the insertion of mini-implants by using two different techniques of site preparation: the superpulsed Er:YAG laser (VPS system) versus the traditional technique with burs. METHODS: One mm wide and 2 mm long implant sites were made in the tibiae of 12 rabbits (8 for each tibia): the authors used an Er:YAG laser with a sapphire tip on a contra-angle hand piece with a 1.0 mm diameter in a VPS mode (with 200 mJ and 30 Hz) to make 4 bone sites and a calibrated traditional bur to drill the other 4 sites in each tibia. The authors inserted a threaded mini-implant, with a sandblasted and acid-etched surface, in the 2 laser implant sites and in the 2 drilled sites of each tibia. The other sites were used as controls. At 0, 7, 15, 30, 45, and 60 days from the surgery, 2 animals each were euthanized, and the retrieved specimens were evaluated. RESULTS: No implant was lost, and the osseointegration was comparable in all implant sites. Histologically, in the 7- and 15-day postoperative specimens, the laser sites showed some amorphous tissue caused by the carbonization of bone during the laser procedure. The sites without implants showed good regeneration of the bone: it was faster in the drill defects because in the laser ones there was some carbonized amorphous tissue. After 30 days postoperatively, the regeneration was the same in all sites. CONCLUSIONS: In comparison with the traditional drilling procedures, Er:YAG laser can be considered efficient in surgery of the bone without inducing irreversible damages, even if it is possible to observe the presence of some carbonized amorphous tissue in the early part of the healing process. This tissue is progressively resorbed in a way similar to a bone graft and doesn't impede the bone formation and osseointegration processes. Further clinical and histological studies are necessary to better define this amorphous tissue and an efficient and safe operative protocol in the surgery of bone with an Er:YAG laser. PMID- 16700655 TI - Bone ingrowths to press-fit and loose-fit implants: comparisons between titanium and hydroxyapatite. AB - The aim of this study was to investigate whether the coating of titanium (Ti) implants with hydroxyapatite (HA) might create a better fixation when titanium implants are implanted into a gap. In each of 16 rats, the medullary cavity of both femurs was entered by an awl from the trochanteric area. With steel burrs it was successively reamed to a diameter of 1.5 mm. In a random manner the proximal part of the cavity in half of the bones was reamed once again to a diameter of 2.0 mm. Nails with a diameter of 1.5 mm and a length of 34 mm were then inserted into the medullary cavity of these bones with press fit at the distal half and a gap to the bone in the proximal half. In the remaining bones the whole medullary canal was reamed to a diameter of 2.0 mm, and nails with a diameter of 2.0 mm and a length of 34 mm were introduced. In all cases, either a pure Ti nail or a Ti nail entirely plasma sprayed with HA was used in a random manner. The surface roughness of the pure Ti was characterized by Ra 2.6 microm and Rt 22 microm. Ra of HA was 7.5 microm and Rt 52 microm. At sacrifice after 16 weeks, both femurs were dissected free from soft tissues and then immersed in fixative. A specimen slice of about 5 mm in thickness was prepared from the subtrochanteric region with a water-cooled band saw. Sample preparation for undecalcified tissue followed the internal guidelines at the laboratories of the Department of Biomaterials/Handicap Research. Generally, bone contact to the nails with HA coating was more predictable than was bone contact to the Ti nails. But due to rather large variations in bone contact between the samples, statistical analyses revealed non-significant differences between the 4 groups (p = 0.083). There were no significant differences between Ti and HA coated nails of 2.0 mm (p = 0.633), nor between Ti and HA coated nails of 1.5 mm (p = 0.924). The pooled values for the 2.0 mm nails showed significantly higher bone bonding contact than the pooled values of the 1.5 mm nails (p = 0.011). Our results, then, indicate that bone bonding contact to implants with a loose fit insertion is less predictable than in press fit insertion, and HA coating seemed to be more predictable than pure Ti. However, due to large variations between the samples, the differences did not reach significant levels. PMID- 16700656 TI - Hydroxyapatite enhances long-term fixation of titanium implants. AB - The aim of this study was to evaluate osseous integration of hydroxyapatite coated titanium implants over time as compared to pure titanium. In 20 rats the medullary cavity of both femoral bones was entered by an awl from the trochanteric area. With steel burrs it was successively reamed to a diameter of 2.0 mm. Nails with a diameter of 2.0 mm and with a length of 34 mm were inserted into the medullary cavity; a pure titanium nail on the left side and a titanium nail entirely plasma-sprayed with hydroxyapatite (HA) on the right side. The surface roughness of the pure titanium was characterized by Ra 2.6 microm and Rt 22 microm, and HA had a roughness of Ra 7.5 (arithmetical mean roughness) microm and Rt (maximum profile height) 52 microm. The rats were randomized to a follow up of 6 and 12 months, respectively. At sacrifice the femoral bones were dissected free from soft tissues. The bones were radiographed and then immersed in fixative. A specimen-slice of about 5 mm thickness was prepared from the region under the trochanter minor with a water cooled band-saw. Sample preparation for undecalcified tissue followed the internal guidelines at the laboratories of Biomaterials/Handicap Research. At 6 months the median bone bonding contact of the implants was 40% (range 0-92) in the titanium group and 34% (0-86) in the HA group. At 12 months the median bone bonding contact was 51% (0-97) in the titanium group and 86% (72-98) in the HA group. In conclusion, we found a significant (p = 0.001) increase in bone bonding contact from 6 to 12 months of the HA coated nails and significantly (p = 0.043) enhanced bone bonding contact in HA coated nails at 12 months as compared to pure titanium nails. PMID- 16700657 TI - Intramedullary fixation of fibular fractures with flexible titanium elastic nails: surgical technique and a case report. AB - Intramedullary fixation of fibular fractures has been reported in the literature. Its advantages include ease of fixation as well as minimal soft tissue disruption. Various implants have been described, including the Inyo nail and Rush rods. Several studies have examined their use in older people with osteoporotic bone as well as in instances where soft tissue preservation is of concern. To our knowledge, no technique has been described using flexible titanium elastic nails (TENS) (Synthes; Paoli, PA). We illustrate such a technique as well as a case report that demonstrates our experience. Insertion of flexible titanium elastic nails requires attention to detail with preservation of the peroneal tendons and their sheath. While such a construct cannot control rotation, it can preserve length as well as prevent varus and valgus displacement. Future endeavors could focus on the biomechanical principles of intramedullary fibular fixation with TENS nails. PMID- 16700658 TI - Manufacturing imperfection of a locking plate screw hole: a case report. AB - Plate and screw fixation devices are essential in the orthopaedic surgical management of various fractures. Production of these devices has been nearly perfected, with few manufacturing defects reported. Assuming that all implants have been machined properly may cause a surgeon to overlook a manufacturing defect and may potentially result in a poor patient outcome. We report a case of the imperfect manufacturing of a less invasive stabilization system (LISS) plate screw hole. This resulted in a loss of fixation of a single locking screw to the plate in the treatment of a distal femoral fracture. Prompt recognition of the defect helped avoid a potential adverse outcome for the patient. When these issues arise, open dialogue with industry is crucial to promptly identify a solution and enhance patient care. This case highlights the manufacturing company's outstanding quality control and prompt solution of such an occurrence. It is the purpose of this report to raise surgeons' awareness of potential manufacturing problems with implants and prevent negative consequences as a result. PMID- 16700659 TI - Trigeminal neuralgia. AB - Trigeminal neuralgia (TN) is the most common facial neuralgia, and is considered to be one of the most painful conditions to affect patients. The rate of occurrence of TN in men and women is 2.5 and 5.7 per 100,000 per year respectively. TN is generally characterized by lancinating, unilateral, paroxysmal pain occurring in the distribution of the fifth cranial nerve. The diagnosis of TN is made clinically by excluding other possible causes of facial pain and is based on signs and symptoms from the patient history such as a trigger zone, typical unilateral lancinating paroxysms following neural disturbance, and a refractory period. Generally, TN can be diagnosed by the typical patient history, a negative neurologic exam, and response to a trial of carbamazepine. Imaging studies should be considered if the diagnosis is uncertain or neurologic abnormalities are noted. Most cases are caused by compression of the trigeminal nerve root, usually within a few millimeters of entry into the pons. In a few cases, TN is caused by a primary demyelinating disorder. The treatment modalities for the management of TN may be divided into medical, surgical, and gamma-knife radiosurgery. Generally, response to drug therapy is good, with over 80% of patients responding to some of the anticonvulsants. Percutaneous approaches to trigeminal gangliolysis are considered to have less associated risk and less cost than open surgical procedures. Three different techniques may be used to perform percutaneous destruction of the ganglion: percutaneous radiofrequency trigeminal gangliolysis (PRTG), percutaneous balloon microcompression (PBM), and percutaneous retrogasserian glycerol rhizotomy (PRGR). Open surgical procedures used in the treatment of TN include microvascular decompression of the trigeminal root and retrogasserian rhizotomy. Additionally, because both of these procedures have greater associated risks, morbidity, and mortality, they are customarily applied only to younger patients in good health. Stereotactic radiosurgery has been established as an alternative treatment for patients who do not respond to optimal medical management. PMID- 16700660 TI - Physical fitness assessment: an update. AB - The American College of Sports Medicine (ACSM) gives the following definition of health-related physical fitness: Physical fitness is defined as a set of attributes that people have or achieve that relates to the ability to perform physical activity. It is also characterized by (1) an ability to perform daily activities with vigor, and (2) a demonstration of traits and capacities that are associated with a low risk of premature development of hypokinetic diseases (e.g., those associated with physical inactivity). Information from an individual's health and medical records can be combined with information from physical fitness assessment to meet the specific health goals and rehabilitative needs of that individual. Attaining adequate informed consent from participants prior to exercise testing is mandatory because of ethical and legal considerations.A physical fitness assessment includes measures of body composition, cardiorespiratory endurance, muscular fitness, and musculoskeletal flexibility. The three common techniques for assessing body composition are hydrostatic weighing, and skinfold measurements, and anthropometric measurements. Cardiorespiratory endurance is a crucial component of physical fitness assessment because of its strong correlation with health and health risks. Maximal oxygen uptake (VO2max) is the traditionally accepted criterion for measuring cardiorespiratory endurance. Although maximal-effort tests must be used to measure VO2max, submaximal exercise can be used to estimate this value. Muscular fitness has historically been used to describe an individual's integrated status of muscular strength and muscular endurance. An individual's muscular strength is specific to a particular muscle or muscle group and refers to the maximal force (N or kg) that the muscle or muscle group can generate. Dynamic strength can be assessed by measuring the movement of an individual's body against an external load. Isokinetic testing may be performed by assessing the muscle tension generated throughout a range of motion at a constant angular velocity. The ability of a muscle group to perform repeated contractions over a specific period of time that is sufficient to cause fatigue is termed muscular endurance. Musculoskeletal flexibility evaluations focus on the joints and associated structures, ligaments, and muscles that cross the joints. The sit-and-reach test and the behind-the-back reach test satisfy many of the criteria for physical assessment of musculoskeletal flexibility. A physical fitness assessment must be integrated into all activities of daily living, as well as the physician's examination, to assess and promote health. PMID- 16700661 TI - Relationships between changes in plasma lipid transfer proteins and apolipoprotein B-100 kinetics during fenofibrate treatment in the metabolic syndrome. AB - The aim of the present study was to investigate the association between changes in apoB (apolipoprotein B-100) kinetics and plasma PLTP (phospholipid transfer protein) and CETP (cholesteryl ester transfer protein) activities in men with MetS (the metabolic syndrome) treated with fenofibrate. Eleven men with MetS underwent a double-blind cross-over treatment with fenofibrate (200 mg/day) or placebo for 5 weeks. Compared with placebo, fenofibrate significantly increased the FCRs (fractional catabolic rates) of apoB in VLDL (very-low-density lipoprotein), IDL (intermediate-density lipoprotein) and LDL (low-density lipoprotein) (all P<0.01), with no significant reduction (-8%; P=0.131) in VLDL apoB PR (production rate), but an almost significant increase (+15%, P=0.061) in LDL-apoB PR. Fenofibrate significantly lowered plasma TG [triacylglycerol (triglyceride); P<0.001], the VLDL-TG/apoB ratio (P=0.003) and CETP activity (P=0.004), but increased plasma HDL (high-density lipoprotein)-cholesterol concentration (P<0.001) and PLTP activity (P=0.03). The increase in PLTP activity was positively associated with the increase in both LDL-apoB FCR (r=0.641, P=0.034) and PR (r=0.625, P=0.040), and this was independent of the fall in plasma CETP activity and lathosterol level. The decrease in CETP activity was positively associated with the decrease in VLDL-apoB PR (r=0.615, P=0.044), but this association was not robust and not independent of changes in PLTP activity and lathosterol levels. Hence, in MetS, the effects of fenofibrate on plasma lipid transfer protein activities, especially PLTP activity, may partially explain the associated changes in apoB kinetics. PMID- 16700663 TI - Immunohistochemical study of cyclooxygenase-2 and p53 expression in skin tumors. AB - Overexpression of cyclooxygenase-2 (COX-2) has been demonstrated in various cancers, including experimentally promoted tumors, gastrointestinal cancers, breast tumors and skin tumors. The mechanism that controls COX-2 expression is not yet clear. Currently, it is reported that COX-2 expression is frequently associated with mutated p53 genes. The goal of this study was to evaluate the expression patterns of COX-2 and p53 in several skin tumors and their correlation. An immunohistochemical method was used to investigate the expression of COX-2 and p53 proteins on formalin-fixed, paraffin-embedded tissue specimens of squamous cell carcinomas (SCC), basal cell carcinomas (BCC), Bowen's disease (BD), actinic keratosis (AK) and porokeratosis. The expression of COX-2 increased in 50% (5/10) of SCC, 80% (8/10) of BCC, 40% (4/10) of BD, 50% (5/10) of AK, and 20% (2/10) of porokeratosis cases. The expression of p53 increased in 90% (9/10) of SCC, 70% (7/10) of BCC, 70% (7/10) of BD, 50% (5/10) of AK, and 40% (4/10) of porokeratosis cases. COX-2 positivity rates of the p53-positive skin tumors were 56%, 100%, 57%, 80% and 25% in SCC, BCC, BD, AK and porokeratosis, respectively. However, the correlation between p53 and COX-2 expression in skin tumors was not statistically significant (P > 0.05). Our results indicate that skin COX-2 and p53 may play roles in skin tumors, but that there is no apparent correlation between the two markers. PMID- 16700662 TI - Genome-wide transcriptional profiling in human squamous cell carcinoma of the skin identifies unique tumor-associated signatures. AB - The elucidation of specific genetic changes associated with human cancer pathogenesis has focused efforts to relate such changes to the neoplastic phenotype. To further our understanding of the genetic basis of human squamous cell carcinoma (SCC) of the skin, this study used a genome-wide (12 627 sequences) approach to determine transcriptional signatures in lesional and nonlesional sites from five SCC patients. Several novel genes involving the p53 pathway, anti-apoptotic pathways, signal transduction, structural loss and DNA replication, including BCL2A1, MUC4, PTPN11 (SHP2) and FGF9, are upregulated in SCC and could warrant further study regarding their role in disease pathogenesis. SCC pathology is likely combinatorial in nature involving the compounded changes from several cellular processes. PMID- 16700664 TI - Anti-CXCR3 staining is useful for detecting human cutaneous and mucosal mast cells. AB - Human synovial mast cells (MC) can be immunolabelled with antihuman CXCR3 antibody (Ab) (clone 49801). We have investigated whether cutaneous and mucosal MC are stained with anti-CXCR3 Ab in paraffin-embedded sections. Immunohistochemical staining and immunofluorescence double staining assays were performed with anti-CXCR3, anti-tryptase, and anti-chymase Ab using normal skin, psoriatic skin lesions, and normal colon. When compared with tryptase and chymase staining, 100% of the cutaneous and 98% of the mucosal MC were positive for CXCR3. Anti-CXCR3 staining is a useful marker for human cutaneous and mucosal MC in paraffin-embedded sections. PMID- 16700665 TI - Detection of mucosal human papilloma virus DNA in bowenoid papulosis, Bowen's disease and squamous cell carcinoma of the skin. AB - Human papilloma virus (HPV) is known to be an etiologic agent for benign warts of the skin. Recently, HPV have been detected in malignant skin and mucosal diseases suggesting that HPV infection can induce malignant skin tumors. In the present study, we examined the presence of mucosal HPV DNA in normal tissue, Bowen's disease (BD), Bowenoid papulosis (BP) and squamous cell carcinoma (SCC) of the skin. We detected the HPV DNA with polymerase chain reactions, and identified the type by DNA sequencing. In the results, we detected HPV DNA in none of the 17 normal controls, two of the three BP (66.7%), one of the 21 BD (4.8%), and six of the 26 SCC of the skin samples (23.0%). The occurrence rates of HPV in BP and SCC were significantly elevated compared to that of normal controls (P < 0.01 and P < 0.01, respectively). In addition, the occurrence rate of HPV in BP was significantly elevated compared to that of BD (P < 0.05). The reproducibility was confirmed with a polymerase chain reaction (PCR) with another primer pair. Of the two cases of BP with positive HPV DNA, one case showed HPV 31 and the other case HPV 16. The case of BD with positive HPV DNA showed HPV 31. Of the six cases of SCC with positive HPV DNA, one case showed HPV 16, another case HPV 34, and the other four cases HPV 31. These results showed that mucosal HPV, including HPV 31 and 16, could be detected in SSC of the skin. Mucosal HPV, not only the epidermodysplasia verruciformis type, appear to induce malignant skin tumors. PMID- 16700666 TI - Narrow-band ultraviolet B as monotherapy and in combination with topical calcipotriol in the treatment of vitiligo. AB - Vitiligo is a common, idiopathic, acquired, depigmenting disease characterized by loss of normal melanin pigments in the skin. The most interesting treatment methods for extensive vitiligo involve psoralen plus ultraviolet A (PUVA) therapy and ultraviolet (UV)-B phototherapy, particularly narrow-band UV-B. In this randomized and comparative study, we investigated the safety and efficacy of narrow band ultraviolet B as monotherapy and in combination with topical calcipotriol in the treatment of generalized vitiligo. Of the 40 vitiligo patients enrolled in the study, 15 were treated with the calcipotriol plus narrow band UV-B (NBUVB) and 25 with narrow band UV-B alone. The patients were randomized into two NBUVB treatment groups. The first group, consisting of 24 patients (all male), received only NBUVB treatment; the second group, consisting of 13 patients (all male) applied 0.05% topical calcipotriol ointments twice daily. Both groups were irradiated with NBUVB (311 nm). In the NBUVB group, the percentage of the body surface affected was reduced from 27.21 +/- 10.41% to 16.25 +/- 8.54% after a mean of 30 treatment sessions. The mean repigmentation percentage was 41.6 +/- 19.4%. In clinical evaluation (moderate and marked/complete response was accepted as an effective treatment), 19 patients (19/24; 79.17%) had clinically good results. In the NBUVB plus calcipotriol group, the percentage of the body surface affected was reduced from 23.35 +/- 6.5% to 13.23 +/- 7.05% after a mean of 30 treatment sessions. The mean repigmentation percentage was 45.01 +/- 19.15%. In clinical evaluation (moderate and marked/complete response was accepted as an effective treatment), 10 patients (10/13; 76.92%) had clinically good results. Statistically significant intragroup reductions from the baseline percentage of the body surface affected were seen at the endpoint of treatment for the two treatment groups (P < 0.001). However, there was no statistically significant difference between the two treatment groups at the end of therapy with respect to the reduction of repigmentation rates (P > 0.05). The present study reconfirmed the efficacy of NBUVB phototherapy in vitiligo. It can be a therapeutic option considered in the management of patients with vitiligo. However, addition of topical calcipotriol to NBUVB did not show any advantage. PMID- 16700667 TI - A 14-year-old girl with lichenoid sarcoidosis successfully treated with tacrolimus. AB - We report a case of lichenoid sarcoidosis in a young girl treated by oral tacrolimus and methylprednisolone. The patient had had a skin eruption from 1 year of age and had developed uveitis at 2 years of age. Her sight had become affected by the uveitis at 8 years of age. When she was 14, she was admitted to the ophthalmology department of our hospital to start treatment with tacrolimus (FK506). She was referred to the department of dermatology for her skin lesions, which were flat, pinkish or normal skin-colored papules scattered on her extremities and the backs of her hands. Upon histology, epithelioid granulomas were seen in the upper dermis and around the erector pili muscles. She received tacrolimus (FK506) 6 mg/day for 3 months for her uveitis. The eye lesions subsided somewhat, and the skin lesions were almost healed after the 3-month course of tacrolimus. However, 4 months after stopping the tacrolimus, her skin and eye lesions relapsed. At that point, she was started on methylprednisolone 16 mg/day for her uveitis. With the methylprednisolone treatment, the inflammation of the eye lesion immediately healed, as did the skin lesions. PMID- 16700668 TI - Successful treatment of diabetic gangrene with topical application of a mixture of peripheral blood mononuclear cells and basic fibroblast growth factor. AB - Diabetic gangrene is a non-healing skin ulcer that is often resistant to most common treatments. It is caused by microvascular disorders and an immunocompromised state which are induced by diabetes mellitus. We report a 65 year-old man with an aggressive, refractory diabetic gangrene on his left foot. Treatment of his diabetic gangrene with topical application of a mixture of peripheral blood mononuclear cells (PBMC) and basic fibroblast growth factor (bFGF) resulted in a dramatic improvement in a short time. The ulcer was completely closed and, in the past 6 months, no new ulceration has been observed. The patient is able to stand and walk by himself. Topical application of a mixture of PBMC and bFGF appears to be a useful, non-invasive and convenient method for the treatment of diabetic gangrene. PMID- 16700669 TI - Aggressive conservative therapy for refractory ulcer with diabetes and/or arteriosclerosis. AB - A foot ulcer due to diabetes and/or arteriosclerosis obliterans (ASO) frequently results in an intractable condition that resists treatment. To cope with this condition, we have developed a combination therapy that includes conventional conservative therapy plus surgical therapy. This aggressive conservative therapy using aggressive debridement, trafermin (Fiblast Spray, Kaken, Japan) treatment and vacuum-assisted closure (VAC) therapy was adopted to treat seven patients suffering from diabetes and ASO-related refractory foot ulcer accompanied by bone exposure. With the exception of one patient who died during the treatment, the remaining six patients obtained limb salvage. The mean time to cure was 8.3 months. This approach should be considered before amputation. Some patients may refuse amputation or cannot tolerate highly invasive surgical treatment including tissue transplantation. In such cases, this aggressive conservative therapy can be employed as a highly useful and reproducible technique requiring simple techniques. PMID- 16700670 TI - Conjunctival squamous cell carcinoma in a patient with hemodialysis-related porphyria cutanea tarda. AB - Hemodialysis-related porphyria cutanea tarda (HRPCT) is characterized by porphyria-like blisters on sun-exposed parts of the face, arms and hands due to elevated porphyrin levels. Conjunctival squamous cell carcinoma (CSCC) is a rare cancer that typically occurs on the sun-damaged ocular surface of elderly male patients. Herein, we report a case of CSCC in a 72-year-old patient who had been on hemodialysis for 8 years, and had been followed up with a diagnosis of HRPCT for 3 years. To our knowledge, this is the first case report documenting an association between HRPCT and CSCC, which may reflect a pathogenetic process connecting them. PMID- 16700671 TI - A case of lymphangitic sporotrichosis occurring on both forearms with a published work review of cases of bilateral sporotrichosis in Japan. AB - The patient, 56-year-old man who was working as a clerk and a farmer, presented with nodules that had appeared on the dorsa of both his hands 3 months earlier. At the first examination, there were multiple dark-red nodules scattered on the fingers, dorsa and wrists of both hands. The nodules were up to 3 cm in diameter and had crusts in the incenters. The patient was suspected to suffer from prurigo and was subsequently treated with topical steroid, but the nodules did not respond. Therefore, a skin biopsy and fungal culture were performed, and the patient was finally diagnosed as having bilateral multiple sporotrichosis. He was then successfully treated with local thermotherapy and oral potassium iodide. Bilaterally-distributed lymphangitic sporotrichosis is very rare and often difficult to diagnose. Careful attention is required to avoid misdiagnosis. PMID- 16700672 TI - A case of discoid lupus erythematosus of the eyelid. AB - A 39-year-old man was first evaluated 12 years ago for erosive erythema of the left lower eyelid. The response to topical therapy was poor. The eyelid lesion was excised on the patient's demand 6 years ago and blepharoplasty was performed. However, recurrence of erythematous plaques at the same site prompted referral to our department. Examination clinically revealed atrophic erythematous plaques and several white papules on the left lower eyelid, in addition to an atrophic erythema of the upper back. The histopathological findings of both plaques and papules were typical of discoid lupus erythematosus. Immunoserological findings suggestive of systemic lupus erythematosus were absent. The lesions improved with administration of low-dose corticosteroids. Discoid lupus erythematosus involving the eyelid is rare, and definitive diagnosis often takes considerable period of time. In patients with lesions of the eyelid such as erythema or papules that are refractory to treatment, skin biopsy and careful physical examination of the entire body for similar lesions are important. PMID- 16700674 TI - Coexistence of nodular and dyshidrosiform pemphigoid. PMID- 16700673 TI - Acitretin induced bipedal edema. AB - Generalized edema has been reported to be a rare side effect of systemic retinoids. We report a patient who developed localized bipedal edema from acitretin. Other causes of bipedal edema were excluded and withdrawal of acitretin led to resolution of the edema. PMID- 16700675 TI - Complete remission of a primary cutaneous follicle-center cell lymphoma (EORTC criteria)/diffuse large B-cell lymphoma (WHO criteria) by single first-line therapy with rituximab. PMID- 16700676 TI - Lipomatous apocrine mixed tumor of the skin associated with chondroid and ossiferous stroma. PMID- 16700678 TI - Subcutaneous lipoma of the index digit presenting as fingertip bulging. PMID- 16700677 TI - Successful treatment of warts with a combination of maxacalcitol ointment and salicylic acid sticking plaster. PMID- 16700679 TI - Do you speak SBAR? PMID- 16700680 TI - A pilot study on the effects of aquatic exercises on discomforts of pregnancy. AB - OBJECTIVES: To estimate the impact of an aquatic exercise program on perception of body image, participation in health-promoting behaviors, barriers to health promoting participation, level of physical discomfort, and mobility. DESIGN: A two-group, quasi-experimental, pretest/posttest design. SAMPLE AND SETTING: A convenience sample of 40 nonexercising pregnant women who were at least 19 weeks gestation were recruited. Sixty percent of the sample was African American. Participants self-selected assignment to either the exercise or nonexercise group. Both groups completed self-report measures and underwent a mobility assessment. The exercise group participated in a 60-minute, 6-week aquatic exercise program three sessions per week. The control group was instructed to continue their normal activities of daily living. RESULTS: Women who had participated in the aquatic exercise program reported significantly less physical discomfort, improved mobility, and improved body image and health-promoting behaviors as compared to control subjects. CONCLUSIONS: Aquatic exercise during pregnancy may enhance physical functioning, decreasing maternal discomfort, improving maternal body image, and improving health-promoting behaviors. PMID- 16700681 TI - Maternal and newborn outcomes related to maternal warming during cesarean delivery. AB - OBJECTIVE: To compare two methods of maternal warming during cesarean delivery under spinal anesthesia on maternal and newborn outcomes. DESIGN: Randomized control trial. SETTING: Two acute care hospitals in central Canada. PATIENTS: 62 women (32 intervention, 30 control). INTERVENTIONS: Women received either a forced-air warming blanket (intervention) or usual care warmed cotton blankets (control). MAIN OUTCOME MEASURES: For mothers: oral temperature, degree of shivering, thermal comfort, and pain scores. For newborns: rectal temperature at birth, 1- and 5-minute Apgar scores, and frequency of interventions for hypoglycemia within 3 hours of birth. RESULTS: With the exception of perceived thermal comfort, women in the two groups were not significantly different in terms of oral temperature, incidence of shivering, and pain scores. Similarly, newborns in both groups were not significantly different in terms of any of the measured variables. Although newborn rectal temperature was within the normal range, mothers in both groups showed a significant decline in body temperature to the mild hypothermic range (control 36.7 +/- 0.4 degrees C to 35.9 +/- 0.5 degrees C, p < .001; intervention 36.8 +/- 0.4 degrees C to 36.1 +/- 0.4 degrees C, p < .001). CONCLUSION: The usual treatment of supplying warmed cotton blankets remains the treatment of choice for this population. PMID- 16700682 TI - A concept analysis of effective breastfeeding. AB - OBJECTIVE: To provide a concept analysis of effective breastfeeding. DATA SOURCES: Medline and CINAHL electronic databases, reference lists from selected articles, and 7 breastfeeding assessment tools. Search terms included the following: breastfeeding and effective, adequate, successful. STUDY SELECTION: Literature was selected according to whether the concept of effective breastfeeding was defined or described. DATA EXTRACTION: Literature was reviewed and data organized as definitions or descriptions, with descriptions grouped into categories sharing similar characteristics. DATA SYNTHESIS: Four characteristics were chosen as essential attributes of effective breastfeeding: positioning, latch, sucking, and milk transfer. CONCLUSIONS: The concept of effective breastfeeding requires further concept development to define and describe effective positioning, latch, sucking, and milk transfer. Future research is needed to determine the relationships between the antecedents, attributes, and consequences of effective breastfeeding. PMID- 16700683 TI - Motivational interviewing to promote sustained breastfeeding. AB - OBJECTIVE: To explore the feasibility of using motivational interviewing to promote sustained breastfeeding by increasing a mother's intent to breastfeed for 6 months and increasing her breastfeeding self-efficacy. DESIGN: A longitudinal experimental two-group design with repeated measures was selected to explore the feasibility of using motivational interviewing to promote sustained breastfeeding in primiparous mothers. SETTING: Three Western rural community hospital sites. PARTICIPANTS: Convenience sample of 73 primiparous breastfeeding mothers ranging between the ages of 19 and 38, M = 25 (SD = 4.5). MAIN OUTCOME MEASURE: Mothers reported the date of their last day of breastfeeding, defined as any breastfeeding during the previous 24-hour period. Breastfeeding behavior was confirmed at each visit by infant test weights. RESULTS: The motivational interviewing group (M = 98.1 days, SD = 75.2) breastfed longer than the comparison group (M= 80.7 days, SD = 71.9); however, this difference was not significant, t(69) = 0.991, p = .325, Cohen's d = 0.24, related to the variability in the sample. CONCLUSIONS: Although not a statistically significant difference, the mean number of days that mothers in the intervention group breastfed was 98 days compared to the mean of 81 days by the comparison group; therefore, motivational interviewing may be useful as a strategy to test in a comprehensive intervention plan. PMID- 16700684 TI - Developing a prenatal breastfeeding workshop to support maternal breastfeeding self-efficacy. AB - A prenatal breastfeeding workshop developed for primiparous women was specifically designed to increase a woman's breastfeeding self-efficacy in the early postpartum period. Research has shown that breastfeeding self-efficacy, defined as a woman's confidence in her ability to breastfeed, is positively related to breastfeeding success. Teaching strategies, based on Bandura's Self Efficacy Theory and adult learning principles, were incorporated in the workshop design. PMID- 16700685 TI - Development of relationship competencies through guided participation for mothers of preterm infants. AB - OBJECTIVE: To explore the effects of a guided participation (GP) intervention on mothers' development of competencies in relating to their preterm infants in neonatal intensive-care units. DESIGN: Mothers were enrolled in a randomized, longitudinal clinical trial to compare the effectiveness of GP with standard care teaching. Baseline data were collected when the infants were 29 weeks postconceptional age. The intervention (GP or standard care teaching) was administered, and relationship competencies were assessed through observation once per week for the next 6 weeks. SETTING: Two neonatal intensive-care units in the Midwest. PARTICIPANTS: A convenience sample of 16 mothers of low-birthweight infants, 8 in each group. INTERVENTIONS: GP of mothers in relating to their infants in the context of caregiving activities was compared with standard care teaching. RESULTS: Mothers in the GP group had expectations and intentions that were more attuned and adaptive to their infants' needs. In addition, the GP group was consistently higher in relationship competencies than the standard care teaching group. CONCLUSIONS: GP can effectively support mothers in developing relationships with their preterm infants in the context of neonatal intensive care unit caregiving activities. PMID- 16700686 TI - Recommendations for sexuality education for early adolescents. AB - OBJECTIVE: To determine community support and identify appropriate learning methodologies, parameters of delivery, and intervention content. DESIGN: Qualitative descriptive study in which participants were interviewed in a semistructured format. Data were analyzed thematically. SETTING: An urban pediatric primary care clinic from which youths and parents were recruited. PARTICIPANTS: Ten youths, 10 parents, and 10 community members. Community members included professional and laypersons who had experience in working with early adolescents or in working with children of any age on sexuality issues. Overall, most participants were female (67%) and African American (67%). RESULTS: Descriptions of early adolescents' knowledge of sexuality, participants' support for sexuality education for early adolescents, recommendations for education content, and preferred methods for education delivery. CONCLUSION: The participants supported comprehensive sexuality education for early adolescents. They believed that it would help youths to be abstinent, would provide some protection from sexual abuse, and would prepare them to practice safer sex in the future. PMID- 16700687 TI - Variability in the phases of the menstrual cycle. AB - OBJECTIVE: To determine variability in the phases of the menstrual cycle among healthy, regularly cycling women. DESIGN: A prospective descriptive study of a new data set with biological markers to estimate parameters of the menstrual cycles. PARTICIPANTS: One hundred forty one healthy women (mean age 29 years) who monitored 3 to 13 menstrual cycles with an electronic fertility monitor and produced 1,060 usable cycles of data. MEASURES AND OUTCOMES: Variability in the length of the menstrual cycle and of the follicular, fertile, and luteal phases, and menses. The estimated day of ovulation and end of the fertile phase was the peak fertility reading on the monitor (i.e., the urinary luteinizing hormone surge). RESULTS: Mean total length was 28.9 days (SD = 3.4) with 95% of the cycles between 22 and 36 days. Intracycle variability of greater than 7 days was observed in 42.5% of the women. Ninety-five percent of the cycles had all 6 days of fertile phase between days 4 and 23, but only 25% of participants had all days of the fertile phase between days 10 and 17. CONCLUSIONS: Among regularly cycling women, there is considerable normal variability in the phases of the menstrual cycle. The follicular phase contributes most to this variability. PMID- 16700688 TI - The lived experience of women diagnosed with polycystic ovary syndrome. AB - OBJECTIVE: To uncover the meaning of living with polycystic ovary syndrome. DESIGN: Phenomenology. SETTING: A women's health care practice in northeast United States. PARTICIPANTS: A purposive sample consisting of 12 women, ages 21 to 48 years, who had been previously diagnosed with polycystic ovary syndrome. DATA COLLECTION: Semistructured interviews. RESULTS: Analysis of the participants' responses revealed the following themes: (a) identifying differences, (b) wanting to be normal, (c) searching for answers, (d) gaining control, (e) attempting to achieve femininity, (f) letting go of guilt, and (g) dealing with it. CONCLUSIONS: Polycystic ovary syndrome is a syndrome that impacts women both physically and psychosocially. Nurses can play a key role in assisting women afflicted with this hormonal disturbance through education and support. PMID- 16700689 TI - Multisystem factors contributing to disparities in preventive health care among lesbian women. AB - Lesbians experience significant health disparities in preventive care utilization and health outcomes compared to heterosexual women. In this study, a multisystems ecologic approach is taken to identify barriers to access, treatment, and preventive care among lesbian women. Recommendations include increasing knowledge of lesbian health care needs, developing cultural sensitivity and competence in communication and care for lesbian women, and creating practice environments that convey respect, acceptance, and welcome to all women, regardless of sexual orientation. PMID- 16700690 TI - Annotated bibliography of NINR findings on women's health in pregnancy and childbirth: 2006 update. PMID- 16700691 TI - Strategies for promoting perinatal patient safety: new ideas and methods to measure success. PMID- 16700692 TI - Perinatal patient safety from the perspective of nurse executives: a round table discussion. AB - Six nurse executives across the United States discussed issues related to perinatal patient safety. Gaps in communication were identified as one of the biggest challenges facing nurse executives. Other issues included expectations of regulators and accreditors, the pressure for productivity with limited resources and staffing, and undercapitalized technology versus safety and staff competence. Each nurse executive discussed a perinatal patient safety initiative implemented recently in her organization. If costs were not an issue, construction of facilities, adoption of electronic documentation, and adding positions to help assure patient safety were at the top of their wish lists. Patient safety continues as the number one priority for nurse executives. PMID- 16700693 TI - The perinatal patient safety nurse: a new role to promote safe care for mothers and babies. AB - Medical malpractice premiums and costs of obstetric claims, settlements, and jury awards are at an all-time high. This article describes one professional liability company's initiative to promote safer perinatal care and decrease costs of claims, including the development of the perinatal patient safety nurse role. The primary responsibility of the perinatal patient safety nurse is to promote safe care for mothers and babies by keeping patient safety as a focus of all unit operations and clinical practices. PMID- 16700694 TI - Using technology to promote perinatal patient safety. AB - Leaders in health care and national health policy recommend information technology information technology as a strategy to promote patient safety. Technology enables error prevention, surveillance, and analysis. Although there is little research about technology and safety in perinatal care, nurses in the specialty can use current evidence about the electronic health record, decision support systems, and medication safety devices to guide practice. This article includes key issues and general recommendations for the use of information technology to promote patient safety, the most common applications relevant to perinatal care, and strategies for perinatal nurses who implement information technology to promote patient safety. PMID- 16700695 TI - Measuring perinatal patient safety: review of current methods. AB - Methods to measure patient safety include structure, process and outcome measures, safety attitude and climate surveys, focus groups, storytelling, executive walk rounds, and external review. Ideally, measures of patient safety should be meaningful, science based, psychometrically sound, feasible, and actionable. Accurate and timely data feedback to caregivers is critical to effect required changes. A balanced set of patient safety measures provides valuable data to guide efforts to improve perinatal patient safety. PMID- 16700696 TI - European Committee on Antimicrobial Susceptibility Testing (EUCAST) Technical Notes on antimicrobial susceptibility testing. AB - The main objectives of the European Committee on Antimicrobial Susceptibility Testing (EUCAST) are to harmonise breakpoints for antimicrobial agents in Europe, and to act as the breakpoint committee for the European Medicines Agency (EMEA) during the registration of new antimicrobial agents. Detailed EUCAST procedures for harmonising and setting breakpoints for antimicrobial agents are available on the EUCAST website. Beginning with the current issue, a series of EUCAST Technical Notes will be published in CMI, based on the rationale documents produced by EUCAST for each of the antimicrobial agents studied, with the aim of highlighting important background information underlying decisions on breakpoints made by EUCAST. PMID- 16700697 TI - Toxoplasma gondii infection in pregnancy: opportunities and pitfalls of serological diagnosis. AB - Because of its life cycle, the recovery of Toxoplasma gondii from biological samples is often impracticable. Consequently, a serological diagnosis represents the first and the most widely used approach to defining the stage of infection. The detection of IgG, IgM, IgA, IgE and IgG avidity by different methods offers this opportunity. However, the results may be affected by difficulties in interpretation, as the same antibody pattern may have a different valency, contingent upon subjects and clinical settings, e.g., pregnant women vs. neonates, and treated vs. untreated patients. This review describes the various factors that should be taken into account when performing serological tests for T. gondii, as well as the pitfalls that may be encountered during the interpretative process. PMID- 16700698 TI - Identification of macrolide-resistant clones of Streptococcus pyogenes in Portugal. AB - Although the overall level of macrolide resistance (27%) has remained stable in Portugal, a rapid inversion in the dominant phenotypes has been noted, with a sharp decrease in the MLS(B) phenotype paralleled by an increase in the M phenotype. To gain further insight into these changes, 325 macrolide-resistant isolates were characterised using a combination of pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The use of Cfr9I, an isoschizomer of SmaI, to digest M phenotype isolates that were refractory to SmaI digestion allowed direct comparison of MLS(B) and M isolates. The results from PFGE and MLST were highly concordant and identified eight major clones, accounting for 92% of the isolates, each of which was associated exclusively with a single macrolide resistance phenotype. Two major clones were found among MLS(B) isolates, characterised by sequence types (ST) 46 (T12/emm22) and ST52 (T28/emm28), whereas clones characterised by ST39 (T4/emm4) and ST28 (T1/emm1) dominated among M isolates. The clone defined by ST52 corresponded to a bacitracin-resistant clone circulating in Europe, and a novel variant expressing other surface antigens (T12/emm22) was detected. The presence of the four major clones has been reported previously in other European countries, suggesting Europe-wide dissemination of a few macrolide-resistant lineages. PMID- 16700699 TI - Streptococcus pneumoniae endophthalmitis: a study of 36 cases with special reference to antibiotic resistance and treatment options. AB - Patients (n = 36) diagnosed with pneumococcal endophthalmitis from six Spanish hospitals between 1986 and 2004 were studied retrospectively. The diagnosis was based on clinical findings, ophthalmological examination, and isolation of Streptococcus pneumoniae from vitreous and/or aqueous humours of 19 patients (definite diagnosis), and from other ocular specimens of 17 patients (probable diagnosis). The mean (+/- SD) age was 69.3 (+/- 16.5) years (range 1.5-89 years), and 20 (55.5%) patients were male. The origin of endophthalmitis was considered exogenous for 34 (94.5%) patients. The most common predisposing factors were previous ocular surgery (n = 25, 69.4%), ocular trauma (n = 5, 13.9%), and close to-eye radiotherapy (n = 3, 8.3%). Eleven (30.5%) patients underwent evisceration as the first therapeutic measure (primary evisceration), and evisceration was performed after antibiotic treatment failure (secondary evisceration) for six (16.7%) patients. Primary evisceration was performed more commonly (63.6%) during 1998-2004, while secondary evisceration was only performed during 1986-1997. Eighteen (50%) patients received intra-vitreous antibiotics (mainly vancomycin), and 31 (86.1%) patients were given systemic antibiotic therapy. The most frequent pneumococcal serogroups isolated were 6, 19, 9, 15 and 23. Pulsed-field gel electrophoresis analysis of 23 isolates revealed that four belonged to the international clones Spain(23F)-1, Spain(6B)-2, Spain(9V)-3 and Sweden(15A)-25. Non-susceptibility rates (i.e., intermediately-resistant and resistant) were: co trimoxazole, 44.8%; penicillin, 33.3%; tetracycline, 31.0%; erythromycin, 21.9%; chloramphenicol, 17.9%; rifampicin, 7.4%; cefotaxime, 5.9%; and levofloxacin, 0%. Although uncommon, pneumococcal endophthalmitis is a medical emergency because of the often aggressive clinical course, poor visual outcome and need for evisceration in a large proportion of patients. PMID- 16700700 TI - A 62-month retrospective epidemiological survey of anaerobic bacteraemia in a university hospital. AB - The incidence of anaerobic bacteraemia was studied retrospectively over 62 months at Mont-Godinne University Hospital, Yvoir, Belgium. The distribution of organisms, clinical presentations, choice of antimicrobial therapy and clinical outcome were analysed. The proportion of positive blood cultures yielding obligate anaerobes was 3.3%. The overall incidence of clinically significant anaerobic bacteraemia was 0.51 cases/1,000 patient admissions (0.61 cases/10,000 hospital-days), but was significantly higher in patients with active haematological malignancies than in other groups (5.97/10,000 vs. 0.33/10,000 hospital-days; p < 0.05). The Bacteroides fragilis group accounted for 61% of isolates, followed by Clostridium spp. (12.2%), Peptostreptococcus spp. and Leptotrichia spp. (7.3% each) and Fusobacterium spp. (4.8%). The most common risk factors were gastrointestinal surgery (49%) and active haematological malignancies with chemotherapy and/or bone marrow graft (47%). One or more co morbidities were present in 30 (77%) of 39 patients. The lower gastrointestinal tract (41%) and the oropharynx (23%) were the two most frequent presumed or proven sources for bacteraemia, with the origin remaining unknown in eight (20.5%) cases. The overall mortality rate (evaluated 7 days after the occurrence of bacteraemia) was 13%. Fatal outcome correlated with the severity of underlying diseases and the immunosuppressed status of the patients rather than with the causative pathogen or the effectiveness of antimicrobial therapy. Likewise, there was no difference in the mortality rate between patients with monomicrobial and polymicrobial bacteraemia. Overall, the data re-emphasise the importance of anaerobic bacteraemia, especially in patients with haematological malignancies. PMID- 16700701 TI - Infections of implantable cardioverter-defibrillators: frequency, predisposing factors and clinical significance. AB - The prognosis for patients with ventricular arrhythmias has improved dramatically with the aid of implantable cardioverter-defibrillators (ICDs). Although infection is a serious complication that frequently causes dysfunction and loss of ICDs, the frequency, predisposing risk-factors, and clinical and microbiological features are only partially understood. This study describes a retrospective review of 423 procedures in 278 patients with ICD primary implants and replacements performed at a tertiary-care hospital. Generators were placed in either a pectoral (68%) or abdominal (32%) site, and electrodes were placed transvenously in 97% of the patients. Most (95%) interventions were performed in a one-stage procedure. Infection developed with ten (2.4%) implanted devices. Four cases occurred within 30 days of surgery ('early infections') and six occurred > 1 month after surgery ('late infections'). In univariate analysis, factors associated with the development of an early infection were: two-stage surgery, a sub-costal approach, and abdominal generator placement. In patients with late infections, a significant association was found with trauma or decubitus ulcer in the generator area. Infection presented with local signs without systemic complications. Seven of the ten patients required complete removal of the system. PMID- 16700702 TI - Investigation of the possible association between nosocomial candiduria and candidaemia. AB - This study aimed to determine whether candiduria is associated with the occurrence of nosocomial candidaemia. In the case-control part of the study, 115 cases (nosocomial candidaemia) and 115 controls (nosocomial bacteraemia) were similar in age, severity of condition and time of hospitalisation. There was a significant association of candidaemia with candiduria (OR 9.79; 95% CI 2.14 44.76). In the microbiology part of the study, 23 pairs of Candida-positive urine and blood cultures were obtained from 23 patients. In ten (43%) cases, the urine and blood culture isolates belonged to different species, and molecular typing showed a difference in two of the 13 cases yielding the same species from both specimens. Overall, there was a significant association between candiduria and candidaemia, but the Candida isolates from urine and blood were different for 52% of the patients. Thus, the data indicated that the urinary tract was probably not a source for the candidaemia. PMID- 16700703 TI - Accuracy of an immune diagnostic assay based on RD1 selected epitopes for active tuberculosis in a clinical setting: a pilot study. AB - A previous case-control study reported that an in-vitro interferon (IFN)-gamma response to early secreted antigenic target (ESAT)-6 selected peptides was associated with active tuberculosis (A-TB). The objective of the present pilot study was to evaluate the diagnostic accuracy of this assay for TB disease in a clinical setting. An IFN-gamma ELISPOT assay was performed on samples from patients with suspected A-TB using two peptides selected from ESAT-6 protein and three peptides selected from culture filtrate 10 (CFP-10) proteins. The results were compared with those obtained by two commercially available assays approved for diagnosis of TB infection (T SPOT-TB and QuantiFERON-TB Gold) which use ESAT 6/CFP-10 (RD1) overlapping peptides. Sensitivity to the RD1 selected peptides was 70% (positive for 16 of 23 patients with microbiologically diagnosed A-TB) and specificity was 91% (positive for three of 32 controls). In contrast, the sensitivity and specificity were 91% and 59%, respectively, for T SPOT-TB, and were 83% and 59%, respectively, for QuantiFERON-TB Gold. The RD1 selected peptides assay had the highest diagnostic odds ratio for A-TB. Thus, the results suggest that an assay based on RD1 selected peptides has a higher diagnostic accuracy for A-TB in a clinical setting compared with commercially available assays based on RD1 overlapping peptides. PMID- 16700704 TI - Analysis of risk-factors among patients with Crimean-Congo haemorrhagic fever virus infection: severity criteria revisited. AB - The aim of this study was to determine the predictors of mortality among patients infected with Crimean-Congo haemorrhagic fever (CCHF) virus. Among patients with acute febrile syndrome, characterised by malaise, bleeding, leukopenia and thrombocytopenia, who were admitted to hospital during the spring and summer of 2002-2004, 54 had positive IgM and/or PCR results for CCHF virus in blood or tissue. The overall case fatality rate was 7.4%. Among the fatalities, haematemesis (p 0.009), melaena (p 0.001) and somnolence (p 0.022) were more common, the median platelet count was significantly lower (10,600/mL vs. 20,000/mL; p 0.038), the mean prothrombin time (27 s vs. 16 s; p 0.002) and mean activated partial thromboplastin time (73 s vs. 44 s; p < 0.001) were longer, and the mean alanine transferase (ALT) level (1,125 vs. 331; p < 0.001), the mean aspartate transferase (AST) level (3,118 vs. 913; p 0.004) and the mean fibrinogen level (119 vs. 340; p 0.012) were higher. Serum IgM and IgG against CCHF virus was detected in 25% and 0%, respectively, of fatal cases, compared with 94% and 62%, respectively, of cases with favourable outcomes. Oral ribavirin was prescribed to 22 (41%) patients. Of the four fatal cases, it was the intention to prescribe ribavirin to three patients, but this was not possible because of haematemesis and melaena. Higher levels of AST (>or= 700 U/L) and ALT (>or= 900 U/L) are suggested for use as severity criteria. Oral ribavirin was not effective for patients with haematemesis, and intravenous ribavirin is necessary for treatment of CCHF. PMID- 16700705 TI - Effect of highly active anti-retroviral therapy and hepatitis C virus co infection on serum levels of pro-inflammatory and immunoregulatory cytokines in human immunodeficiency virus-1-infected individuals. AB - This study aimed to determine the effect of highly active anti-retroviral therapy (HAART) and hepatitis C virus (HCV) co-infection on peripheral levels of interleukin (IL)-2, IL-10, IL-12 (p70), IL-18 and soluble tumour necrosis factor receptor type II (sTNFRII). Serum levels were monitored for a 1-year period in 25 patients infected with human immunodeficiency virus-1 (HIV-1) who were naive for HAART at the initiation of the study, and in four HIV-1-infected long-term non progressors. Serum levels of both IL-18 and sTNFRII at baseline were significantly higher in HIV-1-infected patients than in controls. Baseline levels of IL-18 and sTNFRII were not significantly different in long-term non progressors compared with the other patients. HCV co-infected patients had significantly higher levels of IL-18 and sTNFRII at each time-point compared with patients who were not co-infected with HCV. Irrespective of HCV status, response to HAART resulted in a significant decrease in the levels of both IL-18 and sTNFRII, particularly among patients who achieved HIV viral suppression, but the net decrease observed at the end of follow-up was lower in patients co-infected with HCV. No information was obtained from IL-2, IL-10 and IL-12 (p70) measurements. The data suggest that analysis of serum levels of IL-18 and sTNFRII may be a valuable tool for evaluating the response to HAART, and perhaps for assessing the degree of immune restoration achieved by HAART responders. The results also highlight the relevance of considering the HCV infection status of HIV-1-infected patients in order to avoid misinterpretation of IL-18 and sTNFRII measurements. PMID- 16700706 TI - A large multi-pathogen waterborne community outbreak linked to faecal contamination of a groundwater system, France, 2000. AB - A large waterborne outbreak of infection that occurred during August 2000 in a local community in France was investigated initially via a rapid survey of visits to local physicians. A retrospective cohort study was then conducted on a random cluster sample of residents. Of 709 residents interviewed, 202 (28.5%) were definite cases (at least three liquid stools/day or vomiting) and 62 (8.7%) were probable cases (less than three liquid stools/day or abdominal pain). Those who had drunk tap water had a three-fold increased risk for illness (95% CI 2.4-4.0). The risk increased with the amount of water consumed (chi-square trend: p < 0.0001). Bacteriological analyses of stools were performed for 35 patients and virological analyses for 24 patients. Campylobacter coli, group A rotavirus and norovirus were detected in 31.5%, 71.0% and 21% of samples, respectively. An extensive environmental investigation concluded that a groundwater source to this community had probably been contaminated by agricultural run-off, and a failure in the chlorination system was identified. This is the first documented waterborne outbreak of infection involving human C. coli infections. A better understanding of the factors influencing campylobacter transmission between hosts is required. PMID- 16700707 TI - Development of an internally controlled real-time PCR assay for detection of Chlamydophila psittaci in the LightCycler 2.0 system. AB - A real-time PCR assay with a DNA purification and inhibition control (internal control; IC) was developed to detect Chlamydophila psittaci DNA in human clinical samples. Novel C. psittaci-specific primers targeting the ompA gene were developed. The IC DNA contained the same primer-binding sites and had the same length and nucleotide content as the C. psittaci DNA amplicon, but had a shuffled probe-binding region. The lower limit of detection was 80 target copies/PCR, corresponding to 6,250 copies/mL in a clinical sample. Specificity was tested using reference strains of 30 bacterial species. No amplification was observed from any of these samples. Respiratory samples from eight patients were positive with this PCR. Six of these patients were confirmed as positive for C. psittaci with serological testing. Two patients had increasing antibody titres, but did not fulfil criteria proposed previously for serologically proven Chlamydia spp. infection. The real-time PCR described in this paper is a sensitive, specific and rapid method to detect C. psittaci DNA in human clinical respiratory samples. PMID- 16700708 TI - Detection of Chlamydia trachomatis DNA using MagNA Pure DNA extraction and Cobas Amplicor CT/NG amplification. AB - The automated MagNA Pure DNA extraction method for Chlamydia trachomatis was compared with the manual Cobas Amplicor protocol using 100 microL of input sample volume from 964 specimens. Agreement between protocols was 96.1%. The automated extraction method had a sensitivity of 99% and a specificity of 100%. Amplification inhibition observed after manual preparation of samples (3.8%) was not apparent following automated extraction. Using 200 microL of sample in the automated extraction process lowered the detection limit without raising the inhibition rate. Furthermore, the automated extraction method halved the hands-on time required for the procedure. PMID- 16700709 TI - Characterisation and molecular epidemiology of extended-spectrum beta-lactamase producing Enterobacter cloacae isolated from a district teaching hospital in Taiwan. AB - Enterobacter cloacae (n = 110) isolates from a district hospital in Taiwan were screened for extended-spectrum beta-lactamases (ESBLs). In total, 17 ESBL producers were identified, based on the combination-disk synergy test using cefotaxime and ceftazidime +/- clavulanic acid. Investigation of ESBL genes in 33 ceftazidime-resistant isolates revealed the SHV-12 gene in the same 17 ESBL producers. In addition, one isolate also carried the CTX-M-3 gene, and two isolates also carried the CTX-M-9 gene. No major epidemic clone of ESBL-producers was identified by pulsed-field gel electrophoresis. Routine screening for the ESBL phenotype, focusing on ceftazidime-resistant E. cloacae, should be undertaken in this area. PMID- 16700711 TI - Distribution of the exfoliative toxin D gene in clinical Staphylococcus aureus isolates in France. AB - Exfoliative toxin D (ETD) was identified recently as a new exfoliative toxin serotype. Like other exfoliative toxins, ETD induces intra-epidermal cleavage through the granular layer of the epidermis of neonatal mice. The distribution of ETD production was investigated in Staphylococcus aureus isolates from infected and colonised patients in France. The etd gene was found in 55 (10.5%) of 522 isolates tested. Isolates responsible for bullous impetigo and generalised staphylococcal scalded skin syndrome did not harbour etd, but etd was significantly more frequent in isolates causing cutaneous abscesses and furuncles. Most etd- and Panton-Valentine leukocidin-positive strains belonged to the clone of community-acquired methicillin-resistant S. aureus spreading currently throughout France. PMID- 16700710 TI - CTX-M-15-producing multidrug-resistant enteroaggregative Escherichia coli in the United Arab Emirates. AB - Extended-spectrum beta-lactamase (ESBL) production was demonstrated in five independent, multidrug-resistant isolates of enteroaggregative Escherichia coli (EAEC) from the United Arab Emirates, representing 11.3% of the EAEC isolates recovered during 1 year. All five isolates carried the bla(CTX-M-15) and the bla(TEM-1) genes, the former positioned 48 bp downstream of an ISecp1 element. In two isolates, the bla(CTX-M-15 )and bla(TEM-1) genes were located on a 95-kb plasmid. This is the first detailed description and characterisation of ESBL production in enteroaggregative E. coli and also the first report of CTX-M producing organisms encountered on the Arabian Peninsula. PMID- 16700712 TI - Increased conjugation frequencies in clinical Enterococcus faecium strains harbouring the enterococcal surface protein gene esp. AB - This study compared the in-vitro ability of Enterococcus faecium isolates of different origin to acquire vanA by conjugation in relation to the occurrence of the esp gene. In total, 29 clinical isolates (15/29 esp+), 30 normal intestinal microflora isolates (2/30 esp+) and one probiotic strain (esp-) were studied with a filter-mating assay. Conjugation events were confirmed by PCR and pulsed-field gel electrophoresis. Among the infection-derived isolates, the esp+ isolates had higher conjugation frequencies compared with esp- isolates (p < 0.001), with a median value of 6.4 x 10(-6) transconjugants/donor. The probiotic strain was shown to acquire vanA vancomycin resistance in in-vitro filter mating experiments. PMID- 16700713 TI - Evaluation of Vitek 2 for identification of yeasts in the clinical laboratory. AB - The Vitek 2 system was compared with conventional assimilation, fermentation and morphological methods for its ability to identify yeast isolates from among 151 clinical specimens and 16 known type culture or quality control strains. An unequivocal identification was obtained for 155 (92.8%) isolates, with low discrimination for nine (5.4%) and false identification for three (1.8%) isolates. All isolates of Candida albicans, Candida glabrata and Candida krusei were identified correctly. It was concluded that the Vitek 2 system offers an excellent alternative for the identification of yeasts in a clinical laboratory. PMID- 16700714 TI - A new variant of Brucella melitensis. AB - Brucella melitensis is highly pathogenic and constitutes a serious risk to public health. In Argentina, biovar 1 has been isolated from infected animals, but the Rev.1 strain vaccine is not authorised for use. This report describes nine atypical B. melitensis isolates obtained from humans. These isolates grew slowly, produced small colonies and were susceptible to penicillin and dyes, similar to the B. melitensis Rev.1 vaccine strain, but were inhibited by streptomycin 2.5 mg/L. The isolation of such atypical B. melitensis variants has never been reported from animals in Argentina, and could indicate the emergence of a new mutant variant. PMID- 16700715 TI - Acinetobacter baumannii respiratory isolates in ventilated patients are associated with prolonged hospital stay. PMID- 16700716 TI - Anaerobic bacteraemia in cancer patients. PMID- 16700717 TI - EUCAST Technical Note on daptomycin. PMID- 16700718 TI - 'Gastric' hypoglycaemia--an important concept in diabetes management. PMID- 16700719 TI - Non-cardiac chest pain: an update. AB - Non-cardiac chest pain (NCCP) is very common, affecting up to a quarter of the USA adult population. Recent studies have shown that the disorder has a profound impact on patient's quality of life and is associated with marked increase in utilization of healthcare resources. Non-cardiac chest pain is a heterogeneous disorder with gastrointestinal and non-gastrointestinal causes. After excluding a cardiac cause, most NCCP patients are treated by cardiologists or primary care physicians and only the minority are referred to a gastroenterologist for further work-up. Gastro-oesophageal reflux disease (GORD) is the most common cause for NCCP. The role of oesophageal dysmotility in NCCP has been discounted in recent years. However, visceral hyperalgesia has been shown to play an important role in symptom generation of non-GORD-related NCCP. The main therapeutic interventions in GORD-related NCCP patients are potent antireflux modalities and pain modulators in those with non-GORD-related NCCP. PMID- 16700720 TI - Radial asymmetry of the upper oesophageal sphincter pressure profile: fact or artefact. AB - Radial asymmetry of the upper oesophageal sphincter high-pressure zone (UESHPZ) is an accepted notion based on manometric studies. Our aim was to evaluate the effect of the catheter diameter and configuration on the resting pressure profile of the UES. We studied 14 young (30 +/- 2 years) and 14 healthy elderly volunteers (77 +/- 2 years) using the station pull-through technique. We used a specially designed water perfused manometric assembly that incorporated a proximal round cross-section (4.8 mm) and a distal flat cross-section (4.8 x 1.2 mm). Anterior and posterior manometric sites on the round catheter measured significantly higher pressure values than did the sites oriented laterally at the same level (P < 0.001) in both young and elderly. In contrast, the flat-shaped catheter measured statistically indistinguishable pressures from all four orientations in both age groups. In both young and elderly the anterio-posterior, but not lateral pressures by the round catheter were significantly higher (P < 0.001) than those of the flat catheter. An exaggerated anteriorly and posteriorly oriented pressure may be recorded compared with lateral pressures depending on the diameter and non-conforming shape of the recording catheter with respect to the UES producing the appearance of radial asymmetry in the UESHPZ. PMID- 16700721 TI - Proximal stomach function in obesity with normal or abnormal oesophageal acid exposure. AB - There is an increased prevalence of gastro-oesophageal reflux and symptoms in obese patients. Information about the proximal stomach in obese patients with reflux is lacking. Gastric volume and compliance are similar between obese and lean subjects. To study the proximal stomach function and perception in obese patients with normal or abnormal oesophageal acid exposure, thirty-one obese patients, with normal or abnormal oesophageal acid exposure, underwent medical evaluation of oesophageal and gastrointestinal symptoms by a questionnaire and measurement of proximal stomach function and perception by an electronic barostat and a standardized questionnaire. Nineteen obese patients had abnormal oesophageal acid exposure. The percentage of total time with pH <4 is significantly related to the presence of hiatal hernia, the oesophageal intensity frequency symptom score and gender, i.e. higher percentage in men. The perception cumulative score was significantly different between patients with normal and abnormal oesophageal acid exposure after adjusting for covariates (gender, body mass index, age, minimal distending pressure, gastric tone and gastric compliance). Gastric tone and compliance were significantly related to the perception cumulative score. In conclusion, patients with abnormal oesophageal acid exposure have increased gastric perception. A significant relation among gastric tone, gastric compliance and upper gastrointestinal sensations was shown. PMID- 16700723 TI - Intestinal sensitivity testing by transmucosal electrical nerve stimulation: stimulus parameters to induce conscious perception. AB - Intestinal sensitivity can be tested using transmucosal electrical nerve stimulation. The aim of this study was to establish the stimulus characteristics that determine perception. In six healthy subjects constant current electrical stimuli were applied via an intrajejunal bipolar electrode while measuring perception. Intensity-response tests with stimuli trains of various frequencies (5 and 100 Hz) and pulse durations (50 and 1000 mus) were performed. All stimuli within the broad range tested induced similar-type abdominal sensations, but the intensity of the stimuli to produce perception differed depending on both pulse duration and frequency. A 20-fold increase in pulse duration decreased the intensity of perceived stimuli by a factor of 0.34 +/- 0.04 (P < 0.05); a similar increase in pulse frequency decreased the intensity by a 0.63 +/- 0.07 factor (P < 0.05). When the frequency and duration concomitantly increased, the stimulus intensity decreased by the product of both factors (0.22 +/- 0.04). Transmucosal electrical nerve stimulation of the intestine induces perception within a broad range of stimuli. However, the intensity of the stimuli required to activate sensory pathways is primarily weighted by the duration rather than by the frequency of the pulses. PMID- 16700722 TI - Relationships between hypoglycaemia and gastric emptying abnormalities in insulin treated diabetic patients. AB - We hypothesize that hypoglycaemia in insulin-treated diabetic patients may result from gastric emptying abnormalities causing insulin and food absorption mismatching. We tested gastric emptying in insulin-treated diabetic patients with unexplained hypoglycaemia and without dyspepsia and in diabetic patients without hypoglycaemia, prospectively. Thirty-one diabetic patients with unexplained hypoglycaemic events within 2 h of insulin injection and 18 insulin-treated diabetic patients without hypoglycaemic events underwent gastric emptying breath tests, glycaemic control and autonomic nerve function. Gastric emptying tests were abnormal in 26 (83.9%) and in four (22.2%) patients with and without hypoglycaemia, respectively (P < 0.001). Gastric emptying was significantly slower in hypoglycaemic diabetic patients (t1/2 139.9 +/- 74.1 vs 77.8 +/- 23.3 and t(lag) 95.8 +/- 80.3 vs 32.84 +/- 16.95 min, P < 0.001 for both comparisons; t-tests). A significant association between hypoglycaemic patients and abnormal values of t1/2 and t(lag) was found (P < 0.001). Gastric emptying abnormalities were more frequent in hypoglycaemic patients. We suggest gastric emptying tests for diabetic patients with unexplained hypoglycaemic events. PMID- 16700724 TI - Evidence for a role of inducible nitric oxide synthase in gastric relaxation of mdx mice. AB - Alterations of gastric mechanical activity have been reported in mdx mouse, animal model for Duchenne muscular dystrophy. This study examined if alterations in the vasoactive intestinal polypeptide (VIP) system are present in mdx stomach. Gastric mechanical activity was recorded in vitro as changes of endoluminal pressure and neurally or pharmacologically evoked relaxations were analysed in mdxvs normal stomach. Reverse-transcription polymerase chain reaction was used to detect inducible nitric oxide synthase (iNOS) expression. Relaxations to sodium nitroprusside in mdx stomach showed no difference in comparison with normal preparations. In normal stomach, VIP produced relaxation, which was reduced by VIP6-28, antagonist of VIP receptors, but was not modified by Nomega-nitro-L arginine methyl ester (L-NAME), 1-H-oxodiazol-[1,2,4]-[4,3-a]quinoxaline-1-one (ODQ) or by N-(3-(aminomethyl)-benzyl)acetamidine (1400W) and aminoguanidine, inhibitors of iNOS. In contrast, in mdx stomach VIP responses were antagonized not only by VIP6-28, but also by L-NAME, ODQ, 1400W or aminoguanidine. In normal stomach, the slow relaxation evoked by stimulation at high frequency was reduced by VIP6-28, but it was unaffected by 1400W or aminoguanidine. In mdx stomach, it was reduced by VIP6-28 or 1400W, which did not show additive effects. iNOS mRNA was expressed only in mdx stomach. The results suggest that in mdx gastric preparations, iNOS is functionally expressed, being involved in the slow relaxation induced by VIP. PMID- 16700725 TI - Prokineticin 1 inhibits spontaneous giant contractions in the murine proximal colon through nitric oxide release. AB - Prokineticins are novel peptides with reported effects on gastrointestinal contractility. Prokineticin actions are mediated by distinct prokineticin receptors (PKR1 and PKR2). This study investigated the role of prokineticin 1 in colonic contractility as well as sites of expression of its receptor in the mouse proximal colon by immunohistochemistry and confocal microscopy. Prokineticin 1 suppressed giant contractions in circular muscle. The inhibitory effect of prokineticin 1 on giant contractions was blocked by the nitric oxide synthase (NOS) inhibitor Nomega-nitro-L-arginine methyl ester (L-NAME). In vitro, prokineticin 1 stimulated nitric oxide release from longitudinal muscle-myenteric plexus cultures. This effect was blocked by L-NAME. PKR1 is expressed on myenteric plexus neurons and colocalizes with a small subset of nNOS expressing neurons. This study suggests that PKR1 mediates an inhibitory effect in vitro, most likely through direct or indirect stimulation of nitric oxide release. PKR1 and its natural ligand, prokineticin 1 may be important for modulation of colonic motility. PMID- 16700726 TI - Effects of serotonin transporter inhibition on gastrointestinal motility and colonic sensitivity in the mouse. AB - Serotonin-selective reuptake transporter (SERT) expression is decreased in animal models of intestinal inflammation and in individuals with inflammatory bowel disease (IBD) or irritable bowel syndrome (IBS), and it is possible that resultant changes in intestinal serotonin signalling contribute to the manifestation of clinical features associated with these disorders. The objective of this investigation was to determine whether inhibition of SERT function leads to changes in gut motility and sensitivity. Mice underwent a 14-day treatment with the SERT inhibitor, paroxetine (20 mg kg(-1)), or vehicle (saline/propylene glycol). Gastrointestinal (GI) transit following charcoal gavage, colonic motility, stool frequency and visceromotor responses to colorectal distension were evaluated. In mice treated with paroxetine, stool output was decreased, upper GI transit was delayed, and colonic sensitivity to a nociceptive stimulus was attenuated. These results demonstrate that reduced SERT function (via pharmacological blockade) significantly alters GI motility and sensitivity in mice, and support the concept that altered SERT expression and function could contribute to symptoms associated with IBS and IBD. PMID- 16700727 TI - Magnet Tracking: a new tool for in vivo studies of the rat gastrointestinal motility. AB - Digestive motility was studied in the rat using a miniaturized version of the Magnet Tracking system which monitored the progression of a small magnetic pill through the entire digestive tract. The dynamics of movement was followed and three-dimensional (3-D) images of digestive tract were generated. After a retention period in the stomach and rapid passage through duodenum, the magnet progressed along the small intestine with gradually decreasing speed and longer stationary periods. It remained in the caecum for variable intervals. In the colon, periods of progress alternated with long quiescent periods. Gastric activity oscillated at 5-6 min(-1). In the small intestine, two frequency domains coexisted, showing independent modulations and proximo-distal gradients (40 to >32 and 28 to >20 min(-1)). Caecal oscillations were of 1.5 min(-1). The data allowed the magnet location and calculation of gastric and small intestinal transit times (58 +/- 36 and 83 +/- 14 min respectively), both significantly prolonged by oleate administration (243 +/- 130 and 170 +/- 45 min respectively). Magnet Tracking is a non-invasive tool to study the in vivo spatial and temporal organization of gastrointestinal motility in the rat. PMID- 16700731 TI - Oral transmission of HIV, reality or fiction? An update. AB - Human immunodeficiency virus (HIV) and many other viruses can be isolated in blood and body fluids, including saliva, and can be transmitted by genital genital and especially anal-genital sexual activity. The risk of transmission of HIV via oral sexual practices is very low. Unlike other mucosal areas of the body, the oral cavity appears to be an extremely uncommon transmission route for HIV. We present a review of available evidence on the oral-genital transmission of HIV and analyse the factors that act to protect oral tissues from infection, thereby reducing the risk of HIV transmission by oral sex. Among these factors we highlight the levels of HIV RNA in saliva, presence of fewer CD4+ target cells, presence of IgA antibodies in saliva, presence of other infections in the oral cavity and the endogenous salivary antiviral factors lysozyme, defensins, thrombospondin and secretory leucocyte protease inhibitor (SLPI). PMID- 16700732 TI - Oral mucositis. AB - Mucositis and xerostomia are the most common oral complications of the non surgical therapy of cancer. Mucositis, a common sequel of radio- (DXR), chemo (CXR) and radiochemo-therapy in patients with cancer, or patients requiring haemopoietic stem cell transplants (HSCT), has a direct and significant impact on the quality of life and cost of care, and also affects survival--because of the risk of infection. Apart from dose reduction, preventive and treatment options for mucositis are scarce, although multiple agents have been tested. Evidence suggests that cryotherapy, topical benzydamine and amifostine might provide some benefit in specific situations. The recombinant human keratinocyte growth factor Palifermin (Kepivance) was recently approved as a mucositis intervention in patients receiving conditioning regimens before HSCT for the treatment of haematological malignancies. A number of mechanistically based interventions are in various stages of development. Unfortunately, many other approaches have not been rigorously tested. This paper reviews the clinical features, prevalence, diagnosis, complications, pathogenesis, prophylaxis and management of mucositis. PMID- 16700733 TI - Molecular fingerprinting methods for the discrimination between C. albicans and C. dubliniensis. AB - Opportunistic fungal pathogens are becoming increasingly important causes of both community-acquired and nosocomial infections. The most important fungal pathogens are yeast species belonging to the genus Candida. These species show differences in levels of resistance to antifungal agents and mortality. Consequently, it is important to correctly identify the causative organism to the species level. Identification of Candida dubliniensis in particular remains problematic because of the high degree of phenotypic similarity between this species and Candida albicans. However, as the differences between both are most pronounced at the genetic level, several studies have been conducted in order to provide a specific and rapid identification fingerprinting molecular test. In most candidal infectious, no single DNA fingerprinting technique has evolved as a dominant method, and each method has its advantages, disadvantages and limitations. Moreover, the current challenge of these techniques is to compile standardized patterns in a database for interlaboratory use and future reference. This review provides an overview of most common molecular fingerprinting techniques currently available for discrimination of C. albicans and C. dubliniensis. PMID- 16700734 TI - Oral and perioral herpes simplex virus type 1 (HSV-1) infection: review of its management. AB - Herpes simplex virus type 1 (HSV-1) gives rise to a variety of clinical disorders and is a major cause of morbidity and mortality worldwide. HSV-1 infections are common in oral and perioral area. The aim of the present report was to critically examine the published literature to evaluate the advantages and limitations of therapy of HSV-1 infection in both immunocompetent and immunocompromised patients. Systemic antiviral therapy has been widely accepted as effective for primary herpetic gingivostomatitis. Aciclovir (ACV) 5% cream seems to be the accepted standard topical therapy for herpes labialis, being both effective and well tolerated, although penciclovir 1% cream has been proposed as a potentially useful treatment. Systemic ACV may be effective in reducing the duration of symptoms of recurrent HSV-1 infection, but the optimal timing and dose of the treatment are uncertain. Aciclovir and famciclovir may be of benefit in the acute treatment of severe HSV-1 disease in immunocompromised patients. There is also evidence that prophylactic oral ACV may reduce the frequency and severity of recurrent attack of herpetic infection in immunocompromised patients, but the optimal timing and duration of treatment is uncertain and can vary in different situations. PMID- 16700735 TI - Diversity of gammadelta T cells in patients with Behcet's disease is indicative of polyclonal activation. AB - OBJECTIVE: Behcet's disease (BD) is a multisystemic disease, with vasculitic lesions in the oral and genital mucosa, eyes, joints, skin and brain. We have previously found that gammadelta T cells are increased in peripheral blood of BD patients. The aim of this study was to investigate the extent of gammadelta T cells in oral biopsies from BD patients with special emphasis on the restriction of Vgamma and Vdelta usage. PATIENTS AND METHODS: Expression of Vgamma and Vdelta chains on peripheral blood gammadelta T cells from 31 BD patients and 19 healthy controls was analysed by flow cytometry and the expression of Vgamma and Vdelta chains in nine ulcerated and eight non-ulcerated oral mucosa from BD patients and non-ulcerated oral mucosa from three healthy controls was analysed by immunohistochemistry. RESULTS: Vgamma9 and Vdelta2 were the predominant chains expressed in peripheral blood of BD patients, although other Vgamma and Vdelta chains were also expressed. The presence of gammadelta T cells was only observed in the ulcerated oral mucosa but not in the non-ulcerated mucosa from the BD patients, and not in the non-ulcerated mucosa from the healthy controls. These gammadelta T cells showed no preferential expression of any of the Vgamma or Vdelta chains. CONCLUSION: These data suggest a polyclonal rather than oligoclonal activation of the gammadelta T cells. This may indicate that during repeated inflammation of the oral mucosa, the gammadelta T cells are responding to a wide variety of antigenic stimuli with consequent expansion of gammadelta T cells expressing various Vgamma and Vdelta chains and that different antigenic stimuli or responses may be responsible for the clinical heterogeneity of the disease. PMID- 16700736 TI - Saliva of patients with Type 1 diabetes: effect of smoking on activity of lysosomal exoglycosidases. AB - OBJECTIVE: The aim of our study was to evaluate the influence of smoking on the activity of N-acetyl-beta-hexosaminidase (HEX), its isoenzymes A (HEX-A) and B (HEX-B) and beta-galactosidase (GAL), in the saliva of patients with Type 1 diabetes. METHODS: In the supernatant HEX and its isoenzymes A and B, and beta galactosidase were determined by the method of Chatteriee et al in modification of Zwierz et al (mKat kg(-1) of protein). Protein was determined by the Lowry et al method (mg ml(-1)). RESULTS: The results presented here suggest that diabetes and smoking modify activity of HEX and its isoenzymes, but only combination of diabetes and smoking give a significant increase in the specific activity of HEX and its isoenzymes. CONCLUSIONS: Type 1 diabetes slightly changes the composition of saliva. Smoking cigarettes significantly modifies the composition and properties of saliva in healthy individuals and patients with Type 1 diabetes. PMID- 16700737 TI - RANKL in human periapical granuloma: possible involvement in periapical bone destruction. AB - OBJECTIVES: The cytokine receptor activator of nuclear factor kappaB-ligand (RANKL) has been involved in both the physiological and pathological regulation of osteoclast life span and bone metabolism. Periapical granuloma is a periradicular lesion characterized by periapical bone destruction. The aims of this study were to associate the RANKL mRNA levels to periapical granulomas using the real-time reverse transcriptase-polymerase chain reaction (RT-PCR) technique and to determine the specific cell involved in RANKL synthesis. METHODS: In eight periapical granuloma and eight periodontal ligament samples from periodontally healthy volunteers, RANKL mRNA was detected by real-time RT-PCR. Expression of RANKL on infiltrate leukocytes was further investigated by flow cytometry in six periapical granulomas. RESULTS: Receptor activator of nuclear factor kappaB ligand mRNA levels were higher in periapical granulomas than in healthy periodontal ligament as its RANKL mRNA cycle threshold (Ct) and DeltaCt were significantly lower than that of controls (33.07 +/- 1.24 vs 36.96 +/- 1.69 and 11.58 +/- 3.02 vs 15.60 +/- 3.31, respectively). A 16.2-fold (2.0-131.6) higher RANKL gene expression was detected in the granulomas compared with the control tissues. We determined by flow cytometry that lymphocytes were the predominant leukocyte cells (53.31%), and monocytes and dendritic cells were the main RANKL synthesizers in granuloma lesions. CONCLUSIONS: These data indicate that monocytes synthesized RANKL in periapical granulomas and suggest that RANKL is involved in bone loss associated with periapical lesions. PMID- 16700738 TI - Expression of basement membrane components in odontogenic cysts. AB - OBJECTIVE: To compare the expression of basement membrane components (BMCs), including laminins 1 and 5, collagen type IV, and fibronectin in odontogenic keratocysts (OKCs) with dentigerous cysts (DCs) and radicular cysts (RCs). MATERIALS AND METHODS: Basement membrane components were analysed in 20 OKCs, 20 DCs and 20 RCs using an immunohistochemical technique. RESULTS: Odontogenic keratocysts, DCs and RCs showed positive reaction to all BMCs studied, with different distributions and intensity. OKCs showed continuous linear deposits for laminins 1 and 5 but two staining patterns (continuous and discontinuous) for collagen type IV and fibronectin. DCs exhibited continuous linear deposits for laminins 1 and 5 and collagen type IV but a discontinuous linear deposit for fibronectin. RCs displayed similar results to DCs for laminin 1, collagen type IV and fibronectin. Laminin 5 in RCs had two staining patterns. Constant results in all cysts were strong intensity for laminin 1 and moderate intensity for laminin 5. CONCLUSIONS: Substantial differences in the expression of BMCs among studied cysts were not observed, suggesting that the separation of the epithelial lining in OKCs is not associated with the existence of these proteins. PMID- 16700739 TI - Pilocarpine for the treatment of salivary glands' impairment caused by radioiodine therapy for thyroid cancer. AB - OBJECTIVES: To study the effect of single-dose pilocarpine hydrochloride 5 mg on the whole unstimulated and stimulated salivary flow in patients suffering from thyroid cancer treated with radioiodine therapy, and to investigate the changes in vital signs during the treatment. SUBJECTS AND METHODS: Five such patients were referred with complaints of dry mouth, rampant caries, and pain in the parotid gland region or history of chronic recurrent suppurative sialodenitis. A single dose of 5 mg pilocarpine hydrochloride was administered to each patient and blood pressure, heart rate, body temperature and salivary secretion rate were monitored at 1, 2 and 3 h. RESULTS: A significant elevation of unstimulated and stimulated saliva flow rate was observed in four patients without significant alteration of the monitored vital signs. CONCLUSIONS: Treatment with pilocarpine hydrochloride may be beneficial in the case of impaired salivary function in patients treated with radioiodine. PMID- 16700740 TI - CD10 expression in stromal cells of oral cavity squamous cell carcinoma: a clinic and pathologic correlation. AB - OBJECTIVE: CD10 is expressed on the majority of follicle-center lymphomas and Burkitt lymphomas. CD10 has also been shown to be present in a variety of other neoplasms. DESIGN: The aim of this study was a correlation of CD10 and several parameters: age, tumor size, presence of lymph node metastases, clinic stage, histologic grading, presence of local recurrences. MATERIALS AND METHODS: The tissues of 77 consecutive patients with oral cavity squamous cell carcinoma were evaluated using immunostaining with monoclonal antibody for CD10. MAIN OUTCOME MEASURES: Highly significant correlations were found with the lymph node status, the presence of local recurrences and the histologic grading. The presence of CD10-positive cells was not correlated with the age of patients, tumor size and clinic stage. RESULTS: The results of the present study show that in oral squamous cell carcinoma CD10 positivity is an indicator of worse prognosis. Another strong correlation was found with the presence of local recurrences. Also the histologic grade was significantly correlated with the CD10 positivity. CONCLUSION: Our results point to the fact that CD10 expression can, perhaps, have an important role in tumor invasion, probably facilitating the occurrence of metastases. PMID- 16700741 TI - Effect of deproteinization on composite bond strength in hypocalcified amelogenesis imperfecta. AB - OBJECTIVE: The aim of this study was to evaluate the effect of the treatment of sodium hypochlorite (NaOCl) after acid conditioning of the enamel and dentin of the primary teeth affected with hypocalcified amelogenesis imperfecta (HCAI) on the shear bond strength of the composite material. MATERIALS AND METHODS: Primary teeth from a 12-year-old girl affected with HCAI and primary teeth collected from apparently healthy children were used. A total of four groups, experimental and control with and without NaOCl treatment were specified. In the control group conventional composite procedure was performed and in the treatment group 5% NaOCl was applied after acid conditioning and then the procedure continued as in the control group. RESULTS: In teeth affected with HCAI, enamel shear bond strengths were significantly enhanced in the treatment group compared with the conventional procedure. CONCLUSION: Deproteinization could be attributed as effective in enhancing the enamel bonding in HCAI teeth and could be used to overcome the high failure rates of adhesive restorations in HCAI cases. PMID- 16700742 TI - Oral health in renal transplant recipients administered cyclosporin A or tacrolimus. AB - OBJECTIVE: The aim of this study was to determine the oral status of renal transplant recipients receiving cyclosporin A (CsA) or tacrolimus (FK-506) as immunosuppressant. SUBJECTS AND METHODS: A total of 88 renal transplant recipients receiving CsA (63 men and 25 women, mean age 51.4 years) and 67 receiving FK-506 (57 men and 10 women, mean age 33.5 years) were included in the study. Donor type, histocompatibility, cold ischemia time and prior delayed graft function were similar between the two groups. Demographics and pharmacological data were recorded for all subjects. RESULTS: The results demonstrated that CsA caused a greater number of oral diseases. A greater number of gingival overgrowth was present in patients treated with CsA. However, the combined use with calcium channel blockers increased the gingival overgrowth number. The occurrence of candida in saliva was observed in 80 renal recipients treated with CsA and 20 treated with FK-506. The presence of squamous oral carcinoma (n = 3) and herpes simplex (n = 10) was observed in patients treated with CsA. These alterations were not observed in renal recipients treated with FK-506. CONCLUSIONS: Renal recipients constitute a high-risk group for oral diseases, as they are immunocompromised. However, the FK-506 regime appears to ameliorate this effect, compared with CsA. Adequate pre- and post-transplant oral health care is recommended for these subjects, irrespective of the time interval for which the drug is administered. PMID- 16700743 TI - Platelet-derived growth factor (PDGF) isoform and PDGF receptor expression in drug-induced gingival overgrowth and hereditary gingival fibrosis. AB - OBJECTIVE: To investigate possible associations between platelet-derived growth factor (PDGF), PDGF receptor expression and macrophages in drug-induced and hereditary gingival overgrowth. MATERIALS AND METHODS: Tissues from patients with drug-induced gingival overgrowth (DIGO) (n = 10) and hereditary gingival fibrosis (n = 10) were studied and compared with 'control' gingiva (n = 10). Expression of PDGF and its alpha and beta receptors was investigated immunohistochemically and by RT-PCR. Macrophages were identified by immunostaining for CD68. RESULTS: PDGF isoforms and receptors were detected in most cells within all specimens. There were no differences in the numbers of macrophages, or fibroblasts expressing PDGF or receptors, between groups. The level of PDGF expression by fibroblasts, determined by absorbance measurements, was similar between groups for PDGF A. Significantly lower levels of total PDGF and the receptors were detected in drug induced overgrowth compared to those in hereditary fibrosis (P < 0.004) and control specimens (P < 0.034). All specimens expressed mRNA for PDGF A, PDGF B and alpha and beta receptors. CONCLUSIONS: These data do not support a pivotal role for macrophage-derived PDGF B in the pathogenesis of DIGO. They suggest that fibroblasts in drug-induced lesions have a lowered capacity to produce, and respond to, PDGF, a property not shared by fibroblasts associated with hereditary fibrosis. PMID- 16700744 TI - Pedunculated oncocytic carcinoma in buccal mucosa: immunohistochemical and ultrastructural studies. AB - PURPOSE: In this study we evaluated pedunculated oncocytic carcinoma (OC) in the buccal mucosa via immunohistochemical and ultrastructural studies. PATIENT AND METHODS: An 84-year-old man was referred to our clinic with a pedunculated mass about 4 cm in diameter in the right buccal mucosa. An incision biopsy revealed the diagnosis of oncocytic tumor, and enucleation was performed. The tumor was stained for immunohistochemical analysis using the ABC method and antibodies against cytokeratin (CK), epithelial membrane antigen (EMA), desmin, S-100 protein and muscle-specific actin, respectively. The tumor was stained with uranyl acetate and lead citrate for visualization by electron microscopy. RESULTS: Histopathology results revealed that the tumor consisted of oncocytic cells, characterized by eosinophilic and granular cytoplasm, and atypical nuclei. These cells had infiltrated local blood vessels, salivary glands and muscles. Immunohistochemical staining indicated that these cells were positive for CK and EMA, and negative for desmin, muscle-specific actin and S-100 protein. Electron microscopy revealed numerous dilated cytoplasmic mitochondria, and the cell contours and nucleic shapes of tumor cells were often irregular. CONCLUSIONS: Because the histopathologic features of OC are similar to those of benign oncocytoma, the diagnosis of OC must be confirmed by a combination of clinical and ultrastructural characteristics. PMID- 16700745 TI - Effects of Emdogain on osteoblast gene expression. AB - OBJECTIVE: Emdogain (EMD) is a protein extract purified from porcine enamel and has been introduced in clinical practice to obtain periodontal regeneration. EMD is composed mainly of amelogenins (90%), while the remaining 10% is composed of non-amelogenin enamel matrix proteins such as enamelins, tuftelin, amelin and ameloblastin. Enamel matrix proteins seem to be involved in root formation. EMD has been reported to promote proliferation, migration, adhesion and differentiation of cells associated with healing periodontal tissues in vivo. DESIGN: How this protein acts on osteoblasts is poorly understood. We therefore attempted to address this question by using a microarray technique to identify genes that are differently regulated in osteoblasts exposed to enamel matrix proteins. RESULTS: By using DNA microarrays containing 20,000 genes, we identified several upregulated and downregulated genes in the osteoblast-like cell line (MG-63) cultured with enamel matrix proteins (Emd). The differentially expressed genes cover a broad range of functional activities: (i) signaling transduction, (ii) transcription, (iii) translation, (iv) cell cycle regulation, proliferation and apoptosis, (v) immune system, (vi) vesicular transport and lysosome activity, and (vii) cytoskeleton, cell adhesion and extracellular matrix production. CONCLUSIONS: The data reported are the first genome-wide scan of the effect of enamel matrix proteins on osteoblast-like cells. These results can contribute to our understanding of the molecular mechanisms of bone regeneration and as a model for comparing other materials with similar clinical effects. PMID- 16700746 TI - Smokeless tobacco-induced lamellar body abnormalities. AB - OBJECTIVES: To compare the morphological changes and quantitative distribution of lamellar bodies (Lb) (membrane coating granules) in the hamster cheek pouch epithelium with smokeless tobacco (ST). MATERIALS AND METHODS: Archives of experimental material from previously published studies [S. Ashrafi, A. Das, R. Worawongvasu, B. Mehdinejad and J. Waterhouse (1992) Scanning Microscopy6: 183] were utilized. Animals in experimental group received most ST (snuff) in their right pouch, 5 days weekly, for 24 months, while no snuff was given to control group. After 24 months, the epithelial tissues were processed for electron microscopic study. Volume densities of Lb were assessed by morphometry. MAIN OUTCOME MEASURES: Densities of Lb in the two groups, experimental vs control. RESULTS: In the control, Lb extruded their contents into the intercellular spaces of upper granular layers and in between the last granular cell layers and keratin layers to form a permeability barrier. Conversely, in the smokeless tobacco treated epithelium, the majority of the Lb that were formed remained inside and accumulated within the granular cells, without extruding their contents into the intercellular spaces to form a lipid compound permeability barrier. CONCLUSIONS: Commercial alkaline ST may have contributed to the abnormal accumulation of Lb in the granular cell layer and affected the extrusion process of Lb to form an incomplete permeability barrier in the oral epithelium. PMID- 16700747 TI - Cheilitis as a variation of Candida-associated lesions. AB - OBJECTIVE: Occasionally, the clinical diagnosis of Candida-associated lesion is difficult because of a variety of its clinical manifestations. There have been a few reports on Candida-associated lip lesion except angular cheilitis. In this paper, we investigate the relation of Candida in persistent lesions of the lip. SUBJECTS AND METHODS: The present study includes consecutive nine patients with persistent erosive lip lesion. For each patient the age at presentation, gender, duration of the disease, other symptoms or signs, complete medical history and prior treatment or medications for the symptom were obtained. Then, mycological examination and the direct cytologic examination were performed. RESULTS: The reported average duration of the disease was 7 months. Six of nine patients had received prior treatments with topical steroids at the other clinic, which failed to resolve their symptoms. Six of nine patients had a predisposing factor for candidiasis. In the culture examination, Candida albicans were isolated in seven patients (77.8%). Five of nine patients received the direct cytologic examination, and four of them revealed pseudohyphae of fungi. The anti-fungal treatment was miconazol gel 25 mg four times per day, and average duration of the treatment was 2 weeks. The outcome was as follows: complete remission in five, remarkable response in two and no response in two. CONCLUSION: Our results suggested that one form of the varieties of Candida-associated lesions might be considered in the case of lip lesion with unknown origin that was persistent and ineffective to the topical steroids treatment. PMID- 16700748 TI - Salivary interleukin-6 and tumor necrosis factor-alpha in patients with burning mouth syndrome. AB - Burning mouth syndrome (BMS) is characterized by burning symptoms on the clinically healthy oral mucosa. To date, etiology of BMS is still unknown. We hypothesized that maybe inflammation which is not clinically apparent might lead to burning symptoms which would then result in altered cytokine profile. In the 28 female patients with BMS (age range 48-80 years, mean 64.05 years) and 28 female controls (age range 40-75 years, mean 63.82 years) by use of enzyme-linked immunosorbent assay, interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF alpha) levels were determined. Statistical analysis included use of independent sample t-test and P < 0.05 was considered as significant. Our results show no significant differences between patients and controls regarding salivary IL-6 and TNF-alpha. PMID- 16700751 TI - Evidence and expertise. AB - This paper evaluates attempts to defend established concepts of expertise and clinical judgement against the incursions of evidence-based practice. Two related arguments are considered. The first suggests that standard accounts of evidence based practice imply an overly narrow view of 'evidence', and that a more inclusive concept, incorporating 'patterns of knowing' not recognised by the familiar evidence hierarchies, should be adopted. The second suggests that statistical generalisations cannot be applied non-problematically to individual patients in specific contexts, and points out that this is why we need clinical judgement. In evaluating the first argument, I propose a criterion for what counts as evidence. It is a minimalist criterion but the 'patterns of knowing', referred to in the literature, still fail to meet it. In evaluating the second argument, I will outline the powerful empirical reasons we have for thinking that decisions based on research evidence are usually better than decisions based on clinical judgement; and show that current efforts to rehabilitate clinical judgement seriously underestimate the strength of these reasons. By way of conclusion, I will sketch the ways in which the concept of expertise will have to be modified if we accept evidence-based practice as a template for health-care. PMID- 16700753 TI - Case study: a bridge across the paradigms. AB - Case study as a teaching and research tool has an extensive history in health and social sciences. Despite its suitability for many of the research questions that face nurses, nurses have not fully embraced case study as a comprehensive approach for research. The vagaries of the real-life clinical setting can confound methodologically purist researchers. Case study provides a milieu in which nurse researchers can respond to these vagaries and move towards a paradigmatic openness. In this paper, we argue that case study offers, as yet, under-explored and under-utilised potential as a bridge across the traditional research paradigms. We argue that case study has broad research application and epistemological, ontological and methodological flexibility. When used as a research approach, case study is both the process and end product of research. It provides a delineated boundary for inquiry, and a structural process within which any methods appropriate to investigating a research area can be applied. PMID- 16700752 TI - The potential contribution of emancipatory research methodologies to the field of child health. AB - The knowledge production of researchers interested in improving the health-care of young clients through the employment of emancipatory research methodologies may be limited by the complexity that working with young children presents to the research process. The purpose of this paper is to explore whether emancipatory research methodologies have application within the context of research with children. Critical examination of the challenges inherent in emancipatory research with children reveals that the application of aspects of these approaches presents possibilities for contributing significantly to our knowledge development for the nursing of children. PMID- 16700754 TI - . . . that the social order prevails: death, ritual and the 'Roman' nurse. AB - In this article, the importance of ritual as a collective response to death is discussed. A case example, taken from a larger ethnographic study, is used to explore the responses and reactions of a group of Italian nurses to death as it occurs within an intensive care unit in Rome, Italy. The material presented is used to analyse the significance that cultural, religious and social beliefs and quasi-beliefs can have in nursing practice. The issues highlighted in this examination of the place of ritual in death are located and discussed within their highly specific cultural context and suggest that, where emphasis remains on nurses as a collective rather than on the individual nurse, ritual acts to ensure that social and moral order prevails. PMID- 16700755 TI - Workplace bullying in nursing: towards a more critical organisational perspective. AB - Workplace bullying is a significant issue confronting the nursing profession. Bullying in nursing is frequently described in terms of 'oppressed group' behaviour or 'horizontal violence'. It is proposed that the use of 'oppressed group' behaviour theory has fostered only a partial understanding of the phenomenon in nursing. It is suggested that the continued use of 'oppressed group' behaviour as the major means for understanding bullying in nursing places a flawed emphasis on bullying as a phenomenon that exists only among nurses, rather than considering it within the broader organisational context. The work of Foucault and the 'circuits of power' model proposed by Clegg are used to provide an alternative understanding of the operation of power within organisations and therefore another way to conceive bullying in the nursing workforce. PMID- 16700756 TI - To describe or prescribe: assumptions underlying a prescriptive nursing process approach to spiritual care. AB - Increasing attention is being paid to spirituality in nursing practice. Much of the literature on spiritual care uses the nursing process to describe this aspect of care. However, the use of the nursing process in the area of spirituality may be problematic, depending upon the understandings of the nature and intent of this process. Is it primarily a descriptive process meant to make visible the nursing actions to provide spiritual support, or is it a prescriptive process meant to guide nursing actions for intervening in the spirituality of patients? A prescriptive nursing process approach implies influencing, and in some cases reframing, the spirituality of patients and thereby extends beyond general notions of spiritual support. In this paper we discuss four problematic assumptions that form the basis for a prescriptive approach to spiritual care. We conclude that this approach extends the nursing role beyond appropriate professional boundaries, making it ethically problematic. PMID- 16700757 TI - Postmodern feminist perspectives and nursing research: a passionately interested form of inquiry. AB - The challenges posed by postmodern and poststructural theories profoundly disrupt the certainties of feminist and nursing research, yet at the same time offer possibilities for developing new epistemologies. While there are an increasing number of accounts discussing the theoretical implications of these ideas for nursing research, I wish to discuss the practical and the methodological implications of using postmodern feminist theories within empirical research. In particular, I identify the challenges I encountered through an examination of specific aspects of the research process and through examples drawn from empirical research. I conclude that using postmodern feminist theories requires a continuous engagement with, and interrogation of, the modern epistemological and ontological assumptions of qualitative, feminist nursing research and, in so doing, presents the possibility for nurse scholars to begin to develop a 'passionately interested' methodological approach to nursing inquiry. PMID- 16700758 TI - Transcending transculturalism? Race, ethnicity and health-care. AB - This paper offers a critical commentary on the essentialist concept of ethnicity, which, it is argued, underpins the discourse of transcultural health-care. Following a consideration of the difficulties that ensue from the way in which ethnicity has been theorised within transcultural nursing in particular, the paper turns to a consideration of alternative ways of thinking about ethnicity, which have emerged from more recent social anthropology and postmodernism. It addresses the question of how to therorise ethnicity in a way that does not entail its reification as a set of fixed cultural properties, and makes some tentative suggestions for the possibility of a critical culturalist approach to difference and healthcare practice, which must include a consideration of racisms. PMID- 16700759 TI - Beyond the art of governmentality: unmasking the distributional consequences of health policies. AB - The aim of this article is to critique health policy discourses that are taken for granted. This perspective will allow for the identification of 'exclusionary' health policies, which we define as policies that are thought to offer universal benefit, despite yielding adverse effects for significant groups of people in society. As such, policies that are said to be designed 'for all' frequently benefit only a subset of the population. Our intent is to highlight the distributional consequences of certain health policies that are largely institutionalized in contemporary society. We believe that these distributional effects are explicit representations of power in society and that institutions may provide individual 'choice' and 'freedom' that, in turn, yields separation as an outcome, a separating equilibrium. Specifically, if those who benefit from policies of partition are numerous and are to obtain significant advantage or incur limited costs, or if those who are adversely affected are scarce (or hidden), or the size of these adverse effects are small (or perceived to be minor), then partition becomes more likely as a 'legitimate', but exclusionary, instrument of public policy. PMID- 16700760 TI - Problem-based Learning. PMID- 16700762 TI - Audit or research: the ethics of publication. PMID- 16700761 TI - Career preferences in medicine for the 21st century. PMID- 16700763 TI - Biobanks: can they overcome controversy and deliver on their promise to unravel the origins of common diseases? PMID- 16700764 TI - A covenant model for the medical educator-student relationship: lessons from the covenant model of the physician-patient relationship. PMID- 16700766 TI - Career choices of United Kingdom medical graduates of 2002: questionnaire survey. AB - OBJECTIVES: To report the specialty choices of UK medical graduates of 2002, and to compare their choices with those of qualifiers in previous years and with the profile of career grade doctors in different specialties in England. METHODS: We carried out a postal questionnaire survey in the UK and drew comparisons with official data for numbers of specialists. RESULTS: The response rate was 65.3% (2778/4257). A total of 22.7% of the medical graduates of 2002 (28.1% of women, 14.5% of men) expressed a preference for a longterm career in general practice, compared with 25.3% of 1999 and 2000 graduates combined. A total of 31.1% of men and 11.9% of women chose surgical specialties; 0.7% of men and 3.4% of women chose obstetrics and gynaecology; 3.4% of men and 7.9% of women chose paediatrics. There was a large mismatch between the percentage choosing each specialty group and the percentage of senior National Health Service doctors working in the same specialty group. In all, 71% of graduates regarded their career choice as definite or probable and 80% definitely or probably intended to pursue a longterm career in medicine in the UK, compared with 75% of qualifiers in 1999 and 2000. CONCLUSIONS: Career choices for general practice remain low: only 1 in 4 women and 1 in 7 men now choose general practice at this career stage. Very few men choose obstetrics and gynaecology, despite a recent increase in training opportunities. There is no evidence of an increase, compared with recent previous cohorts, in the percentage of junior doctors who do not want a longterm career in British medicine. PMID- 16700765 TI - Interdisciplinary health education and career choice in rural and underserved areas. AB - PURPOSE: To assess the association between an educational rural health interdisciplinary programme (RHIP) and subsequent practice in US rural and underserved locations. METHODS: We carried out a longitudinal cohort study of RHIP students and randomly selected classmate controls for the years 1990-2001, using a mailed survey. OUTCOMES: Main outcome measures were first rural, any rural, first underserved and any underserved practice locations. Multivariate statistical methods were used to calculate prevalence ratios (PRs) by discipline while controlling for possible extraneous variables. RESULTS: Of 1396 surveys delivered, 820 were returned, giving a response rate of 59%. After exclusions, results from 255 RHIP and 534 control students were analysed for outcomes. Pharmacy students on the RHIP chose first and any rural practice locations more often than reference controls (PRs = 2.59 and 1.97, respectively; P < 0.05). Therapies (occupational, physical and speech therapy) RHIP students were associated with all 4 practice outcomes more often (PRs = 2.07, 1.85, 1.68 and 1.65, respectively; P < 0.05). Pharmacy and Therapies control students with rural training chose first rural and any rural practices more often (PRs = 2.58 and 1.62, respectively; P < 0.05 for both). Medicine and Nursing students did not choose outcome practice locations more often, but had small sample sizes and large numbers of controls with rural training. Rural health interdisciplinary students rated participation in rural training more highly as a factor in choosing first rural practices than did the controls who chose similar practices. CONCLUSIONS: Participation in RHIP and other rural training experiences may stimulate subsequent career choices in rural and underserved locations for Pharmacy and Therapies students. Other studies are needed to confirm these findings and answer questions raised by these data. PMID- 16700767 TI - Influences on choice of surgery as a career: a study of consecutive cohorts in a medical school. AB - AIM: To examine the differential impact of person-based and programme-related features on graduates' dichotomous choice between surgical or non-surgical field specialties for first-year residency. METHODS: A 10-year cohort study was conducted, following 578 students (55.4% male) who graduated from a university medical school during 1994-2003. Data were collected as follows: at the beginning of medical studies, on career preference and learning frame; during medical studies, on academic achievement, cross-year peer tutoring and selective clinical traineeship, and at graduation, on the first-year residency selected. Contingency and logistic regression analyses were performed, with graduates grouped by the dichotomous choice of surgery or not. RESULTS: Overall, 23% of graduates selected a first-year residency in surgery. Seven time-steady features related to this choice: male sex, high self-confidence, option of surgery at admission, active learning style, preference for surgery after Year 1, peer tutoring on clinical surgery, and selective training in clinical surgery. Logistic regression analysis, including all features, predicted 87.1% of the graduates' choices. Male sex, updated preference, peer tutoring and selective training were the most significant predictors in the pathway to choice. DISCUSSION: The relative roles of person-based and programme-related factors in the choice process are discussed. The findings suggest that for most students the choice of surgery derives from a temporal summation of influences that encompass entry and post entry factors blended in variable patterns. It is likely that sex-unbiased peer tutoring and selective training supported the students' search process for personal compatibility with specialty-related domains of content and process. PMID- 16700768 TI - The compatibility of future doctors' career intentions with changing health care demands. AB - BACKGROUND: In the Netherlands the medical education system is in the process of being transformed to establish a more demand-oriented health care system. This transformation may entail the occupational restructuring of the medical profession. Meanwhile, on the supply side, the career intentions of future doctors are also changing. OBJECTIVES: We aimed to categorise medical students' prevailing career intentions and to examine to what extent newly proposed medical occupations that may be part of the transformation process correspond with these career intentions. METHODS: We carried out expert interviews and a feedback round to gain input for a survey among students. From the demand perspective, 11 experts proposed non-traditional medical occupations and evaluated these on the basis of job characteristics relevant to a doctor's career choice. Subsequently, students from 5 universities filled out a questionnaire to rate these job characteristics by their importance and the proposed occupations' attractiveness. Results Four different clusters of career intentions were categorised as patient oriented expert, career-oriented specialist, lifestyle-oriented generalist, and balance-seeking realist. These clusters differ in terms of the ways in which students feel attracted to the proposed occupations. The career-oriented specialists feel least attracted and the lifestyle-oriented generalists most attracted to the occupations. DISCUSSION: The experts' call for shorter postgraduate programmes to educate patient-oriented doctors partly matches students' career intentions. Most students share the intention of obtaining a direct care position that provides ample task variation, which may explain the appeal of the occupations 'emergency doctor' and 'basic specialist'. The limited interest in specific patient groups suggests a need for more exposure to the occupations linked to these groups. PMID- 16700769 TI - How well do paediatric residency programmes prepare residents for clinical practice and their future careers? AB - CONTEXT: Educators across Canada are presently discussing whether the current 4 year residency programmes adequately prepare paediatricians for their future careers. Studies carried out in the USA have repeatedly shown areas of weakness in residency training, but there are no studies looking at the overall adequacy of training across Canada. OBJECTIVES: To assess practising paediatricians' perceptions of the adequacy of their residency training as preparation for clinical practice and to assess practising paediatricians' opinions about the required mandatory length of training. METHODS: A questionnaire based on previous studies was sent to 434 paediatricians certified between 1999 and 2003, asking for their opinions of their preparedness for practice in the broad areas of paediatrics and in the professional roles of the doctor-specialist. RESULTS: Overall, 239 (55%) paediatricians replied, 96% of whom indicated they were 'adequately' or 'very well' trained. Areas in which opinions on training were positive included emergency medicine, neonatology, endocrinology, haematology/oncology, neurology, infectious diseases and respirology. Areas where preparation was considered to have been less adequate included gynaecology, child psychiatry, behavioural psychology, surgical specialties, orthopaedics and adolescents. With respect to the roles of the doctor-specialist, strengths of training included the areas of medical expert, collaborator, ethics and professionalism, and communicator. Respondents felt they were less adequately prepared for the role of a medical expert dealing with palliative care, for dealing with bereaved parents and as manager of an office practice. Despite these weaknesses, 80% felt that 4 years of training was sufficient. DISCUSSION: The results of the study are comparable with those of previous studies carried out in the USA and reinforce the need for regular programme assessment. This study will hopefully lead to the improvement of current paediatric residency programmes and enhanced education and training of future paediatricians. Although overall satisfaction with training was high, paediatric programmes need to make some changes by providing more appropriate training with less tertiary care, hospital based training and more community and ambulatory-based experiences. PMID- 16700770 TI - A comparison of the effects of problem-based learning and lecturing on the development of students' critical thinking. AB - BACKGROUND: Educational approaches are thought to have facilitative or hindering effects on students' critical thinking development. The aim of this study was to compare the effects of problem-based learning (PBL) and lecturing approaches on the development of students' critical thinking. METHODS: All 79 Year 1 undergraduate nursing students at a university in Hong Kong were randomly assigned to 1 of 2 parallel courses delivered by either PBL (n = 40) or lecturing (n = 39) over 1 academic year. The primary outcome measure was students' critical thinking disposition as measured by the California Critical Thinking Disposition Inventory (CCTDI). Individual interviews were also conducted to elicit the students' perceptions of their learning experience. Data were collected at 4 timepoints spanning 3 years. RESULTS: The overall CCTDI and subscale scores for the PBL group were not significantly different from those of the lecture group at the first timepoint (pretest). Compared with lecture students, PBL students showed significantly greater improvement in overall CCTDI (P = 0.0048), Truthseeking (P = 0.0008), Analyticity (P = 0.0368) and Critical Thinking Self confidence (P = 0.0342) subscale scores from the first to the second timepoints; in overall CCTDI (P = 0.0083), Truthseeking (P = 0.0090) and Analyticity (P = 0.0354) subscale scores from the first to the third timepoints; and in Truthseeking (P = 0.0173) and Systematicity (P = 0.0440) subscale scores from the first to the fourth timepoints. CONCLUSIONS: There were significant differences in the development of students' critical thinking dispositions between those who undertook the PBL and lecture courses, respectively. PMID- 16700771 TI - Measuring the effect of interprofessional problem-based learning on the attitudes of undergraduate health care students. AB - OBJECTIVES: This study aimed to measure the improvement in attitudes towards interprofessional collaboration of undergraduate health care students who have a single module of interprofessional problem-based learning (PBL) using real patients as triggers integrated into their curricula. DESIGN: A dedicated module, consisting of 5 PBL seminars, was integrated into the undergraduate medical, nursing and physiotherapy curricula at the participating institutions. Seminar groups consisted of students from a single profession in the control group, and of evenly distributed students from the participating professions in the intervention group. The Interdisciplinary Education Perception Scale was used to measure improvements in attitudes towards interprofessional co-operation. Patients, faculty members and students were included in the evaluation of the interprofessional module and their comments examined for indications of adverse effects of the use of patients in this setting. RESULTS: A total of 177 students were recruited into the study and assigned to 1 of 16 seminar groups, all of which attended the complete module, 8 in control mode and 8 in intervention mode. Statistically significant improvements could be identified in the overall attitudes of male students in the intervention group, and in attitudes pertaining to the competence and autonomy of individuals in one's own profession in the intervention group as a whole. No significant improvements were detected in the control group. No adverse effects of the use of real patients came to light. CONCLUSIONS: The integration of an interprofessional educational module that requires limited student and faculty time in undergraduate health care curricula may be proven to have an effect. The Interdisciplinary Education Perception Scale may be suitable for measuring such effect. Real patients may continue to contribute to education in this setting. PMID- 16700772 TI - Longterm effects of problem-based learning: a comparison of competencies acquired by graduates of a problem-based and a conventional medical school. AB - BACKGROUND: Problem-based learning (PBL) as an approach to the instruction of medical students has attracted much attention in recent years. However, its effect on the performance of its graduates is the subject of considerable debate. This article presents data from a large-scale study among graduates of a problem based medical school and those of a conventional medical school to contribute to this discussion. PURPOSE: To study the longterm effects of problem-based medical training on the professional competencies of graduates. METHODS: A questionnaire was sent to all graduates since 1980 of a problem-based and a conventional medical school. Participants were requested to rate themselves on 18 professional competencies derived from the literature. RESULTS: The graduates of the PBL school scored higher on 14 of 18 professional competencies. Graduates of the problem-based school rated themselves as having much better interpersonal skills, better competencies in problem solving, self-directed learning and information gathering, and somewhat better task-supporting skills, such as the ability to work and plan efficiently. There were no sizeable differences with regard to general academic competencies, such as conducting research or writing a paper. Graduates from the conventional school rated themselves as having slightly more medical knowledge. The findings were shown to be valid and robust against possible response bias. CONCLUSION: The findings suggest that PBL not only affects the typical PBL-related competencies in the interpersonal and cognitive domains, but also the more general work-related skills that are deemed important for success in professional practice. PMID- 16700773 TI - Student perceptions of a virtual learning environment for a problem-based learning undergraduate medical curriculum. AB - OBJECTIVE: To investigate the effects of a virtual learning environment (VLE) on group interaction and consultation of information resources during the preliminary phase, self-study phase and reporting phase of the problem-based learning process in an undergraduate medical curriculum. METHODS: A questionnaire was administered to 355 medical students in Years 1 and 2 to ask them about the perceived usefulness of a virtual learning environment that was created with Blackboard for group interaction and the use of learning resources. RESULTS: The students indicated that the VLE supported face-to-face interaction in the preliminary discussion and in the reporting phase but did not stimulate computer mediated distance interaction during the self-study phase. They perceived that the use of multimedia in case presentations led to a better quality of group discussion than if case presentations were exclusively text-based. They also indicated that the information resources that were hyperlinked in the VLE stimulated the consultation of these resources during self-study, but not during the reporting phase. CONCLUSIONS: Students indicated that the use of a VLE in the tutorial room and the inclusion of multimedia in case presentations supported processes of active learning in the tutorial groups. However, if we want to exploit the full potential of asynchronous computer-mediated communication to initiate in-depth discussion during the self-study phase, its application will have to be selective and deliberate. Students indicated that the links in the VLE to selected information in library repositories supported their learning. PMID- 16700774 TI - Validation of learning style measures: implications for medical education practice. AB - BACKGROUND: It is unclear which learners would most benefit from the more individualised, student-structured, interactive approaches characteristic of problem-based and computer-assisted learning. The validity of learning style measures is uncertain, and there is no unifying learning style construct identified to predict such learners. OBJECTIVE: This study was conducted to validate learning style constructs and to identify the learners most likely to benefit from problem-based and computer-assisted curricula. METHODS: Using a cross-sectional design, 3 established learning style inventories were administered to 97 post-Year 2 medical students. Cognitive personality was measured by the Group Embedded Figures Test, information processing by the Learning Styles Inventory, and instructional preference by the Learning Preference Inventory. The 11 subscales from the 3 inventories were factor analysed to identify common learning constructs and to verify construct validity. Concurrent validity was determined by intercorrelations of the 11 subscales. RESULTS: A total of 94 pre-clinical medical students completed all 3 inventories. Five meaningful learning style constructs were derived from the 11 subscales: student- versus teacher-structured learning; concrete versus abstract learning; passive versus active learning; individual versus group learning, and field dependence versus field-independence. The concurrent validity of 10 of 11 subscales was supported by correlation analysis. Medical students most likely to thrive in a problem-based or computer-assisted learning environment would be expected to score highly on abstract, active and individual learning constructs and would be more field-independent. CONCLUSIONS: Learning style measures were validated in a medical student population and learning constructs were established for identifying learners who would most likely benefit from a problem based or computer-assisted curriculum. PMID- 16700775 TI - Stress, debt and undergraduate medical student performance. AB - INTRODUCTION: Against the background of current debate over university funding and widening access, we aimed to examine the relationships between student debt, mental health and academic performance. METHODS: We carried out an electronic survey of all medical undergraduate students at the University of Aberdeen during May-June 2004. The questionnaire contained items about demographics, debt, income and stress. Students were also asked for consent to access their examination results, which were correlated with their answers. Statistical analyses of the relationships between debt, performance and stress were performed. RESULTS: The median total outstanding debt was pound 7300 (interquartile range 2000-14 762.50). Students from lower socioeconomic backgrounds and postgraduate students had higher debts. There was no direct correlation between debt, class ranking or General Health Questionnaire (GHQ) score; however, a subgroup of 125 students (37.7%), who said that worrying about money affected their studies, did have higher debt and were ranked lower in their classes. Some of these students were also cases on the GHQ-12. Overall, however, cases on the GHQ had lower levels of debt and lower class ranking, suggesting that financial worries are only 1 cause of mental health difficulties. DISCUSSION: Students' perceptions of their own levels of debt rather than level of debt per se relates to performance. Students who worry about money have higher debts and perform less well than their peers in degree examinations. Some students in this subgroup were also identified by the GHQ and may have mental health problems. The relationships between debt, mental health and performance in undergraduate medical students are complex but need to be appreciated by medical education policy makers. PMID- 16700776 TI - Ethical decision making on truth telling in terminal cancer: medical students' choices between patient autonomy and family paternalism. AB - INTRODUCTION: The effect of medical education is often hard to evaluate. We tried to assess whether a 2-hour, small-group discussion could alter student perspectives on truth telling. Currently, in Taiwan it is common practice to consult with the family of a terminally ill patient before telling the truth to the patient, which may be in conflict with patient autonomy. METHODS: The study was based on content analysis of self-reflective written texts after a 2-hour group discussion on a clinical case describing a truth-telling situation. The changes in decision patterns regarding the emphasis placed on patient autonomy versus family paternalism and the connection to related moral reasoning were subjected to focus analysis. RESULTS: The students' initial attitudes regarding the subject of truth telling were categorised into 4 patterns, namely, patient centred (n = 46), family-centred (n = 20), simultaneous informing (n = 1), and situational informing (n = 5) modes. The discussion stimulated perspective changes in many of the students and their attitudes were then regrouped as patient-comprehensive (n = 35), family-centred (n = 1), and family-comprehensive (n = 36) modes. It was found that variations on 'common sense' and moral reasoning existed prior to the class and the students initially tended to overlook the complexity of truth telling in terminal cancer. Through the discussion and reflective learning, they were enabled to acknowledge the vulnerability of both the patient and his or her family, and to make decisions based on more comprehensive considerations. CONCLUSION: Group discussion seemed to be able to enhance ethical consideration. Further research is required to determine whether the benefits of this approach can be translated into behavioural changes in practice. PMID- 16700777 TI - The assessment of professionalism. PMID- 16700778 TI - Mini-clinEx versus mini-CEX. PMID- 16700779 TI - Current modalities and new advances in the treatment of basal cell carcinoma. AB - Basal cell carcinoma (BCC) is one of the most common cancers. Surgical extirpation is currently the standard of care for BCC, which is associated with several advantages and disadvantages. Procedures such as surgical excision used to treat superficial BCC (sBCC) and nodular BCC (nBCC) may have high 5-year recurrence rates if tumors are not completely excised. Curettage with electrodesiccation is a common method for treating primary BCC. However, multiple cycles are recommended and the procedure can have unsatisfactory cosmetic results (e.g. scarring and hypopigmentation). Mohs micrographic surgery has a low rate of disease recurrence but is a specialized procedure usually limited to specific indications (e.g. high-risk tumors). Cryosurgery and photodynamic therapy require multiple cycles and are associated with variable cosmetic outcomes and recurrence rates. As with any procedure, potential risks and patient quality-of-life issues need to be considered. In addition, substantial patient and healthcare provider inconvenience limit the practical utility of some modalities. Pharmacologic interventions provide another treatment option as adjunctive or monotherapy. Investigations of imiquimod, a novel immune response modifier, have indicated that this topical, noninvasive agent is safe and well tolerated and may be efficacious in the treatment of BCC. This review will highlight the role of standard treatment modalities and introduce new advances in the treatment of BCC. PMID- 16700780 TI - Angiosarcoma arising in a lymphedematous abdominal pannus with histologic features reminiscent of Kaposi's sarcoma: report of a case and review of the literature. AB - BACKGROUND: Despite common endothelial origins, angiosarcoma and Kaposi's sarcoma are clinically and histologically distinct vascular proliferations. The development of angiosarcoma in a chronically edematous abdominal pannus is extremely uncommon. Similarly, tumors with the combined histologic features of angiosarcoma and Kaposi's sarcoma have rarely been described. METHODS: We reviewed the literature on angiosarcoma arising in a lymphedematous abdominal pannus and evaluated an 81-year-old morbidly obese woman who had profound, long standing edema of the lower abdominal wall in which an aggressive vascular tumor developed. RESULTS: Three clinically similar cases were identified in the literature. All patients were women who generally experienced rapid disease progression. In addition, in our patient, sequential cutaneous sampling from different lesional sites demonstrated disparate histologic changes, ranging from those of classic Kaposi's sarcoma to high-grade angiosarcoma, to areas with combined features of the two tumors. A polymerase chain reaction performed on lesional tissue was negative for human herpesvirus-8 DNA. CONCLUSION: It is important to note that angiosarcoma may develop in the abdomen in association with chronic lymphedema, as demonstrated by the cases noted in this report. In addition, our case highlights the difficulty in differentiating histologically angiosarcoma from Kaposi's sarcoma in some situations, and demonstrates the value of close clinicopathologic correlation and sequential tissue sampling in evaluating problematic cases. PMID- 16700782 TI - Lupus erythematosus tumidus: a series of 26 cases. AB - OBJECTIVE: To study 26 cases of lupus erythematosus tumidus (LET), a subset of chronic cutaneous lupus erythematosus (CCLE), referred to in the literature as a rare entity. PATIENTS AND METHODS: A retrospective study was conducted of 26 patients diagnosed with LET between 1996 and 2002. The clinical characteristics, histopathologic and laboratory findings, response to treatment, association with other subsets of lupus, course, and diagnostic criteria were analyzed. RESULTS: The incidence by sex was similar. The mean age of presentation was 49.19 years. The clinical presentation usually involved erythematous, edematous plaques located on the face, chest, back, or extremities, related to sun exposure. A dermal lymphocytic infiltrate with a perivascular disposition and differing degrees of mucin deposition was observed in all cases. Minimal epidermal changes were present in 18 cases, and 11 of these also showed minimal dermal-epidermal changes. Only one case showed dermal-epidermal changes without any epidermal alteration. Direct immunofluorescence test was performed in 15 patients, and 11 were negative. All cases showed a benign course without systemic manifestations. The response to topical steroids or antimalarial treatment was excellent, but a seasonal recurrence was usually observed. Discussion No defined criteria for LET are universally accepted. The main controversies are the acceptance of LET as a separate subset of CCLE, and the histopathologic diagnostic features, mainly the presence or absence of epidermal and dermal-epidermal changes in these lesions. CONCLUSIONS: No inflexible histologic criteria should be employed for the diagnosis of LET. This subset of lupus erythematosus is characterized by intense photosensitivity, definite clinical lesions, a benign course, the absence of systemic disease, good response to antimalarial treatment, and a tendency to recur. More studies should be performed in order to establish the true incidence of LET because this subset of CCLE is probably underestimated. PMID- 16700781 TI - Antimicrobial properties of Dead Sea black mineral mud. AB - BACKGROUND: The unique, black, hypersaline mud mined from the Dead Sea shores is extensively used in mud packs, masks, and topical body and facial treatments in spas surrounding the lake, and in cosmetic preparations marketed worldwide, but little is known about its antimicrobiological properties. METHODS: We performed detailed microbial and chemical analysis of Dead Sea mineral mud compounded in dermatological and cosmetic preparations. RESULTS: Using conventional bacteriological media (with or without salt augmentation), we found surprisingly low numbers of colony-forming microorganisms in the mud. The highest counts (up to 20,000 colonies per gram, mostly consisting of endospore-forming bacteria) were obtained on sheep blood agar. Test microorganisms (i.e. Escherichia coli, Staphylococcus aureus, Propionibacterium acnes, Candida albicans) rapidly lost their viability when added to the mud. Zones of growth inhibition were observed around discs of Dead Sea mud placed on agar plates inoculated with Candida or with Propionibacterium, but not with Staphylococcus or Escherichia. The effect was also found when the mud was sterilized by gamma irradiation. Using (35)S labeled sulfate as a tracer, bacterial dissimilatory sulfate reduction could be demonstrated at a low rate (0.13 +/- 0.03 nmol/cm(3).d). CONCLUSION: The antibacterial properties of Dead Sea mud are probably owing to chemical and/or physical phenomena. Possible modes of antimicrobial action of the mud in relation to its therapeutic properties are discussed. PMID- 16700783 TI - Evaluation of desmoglein enzyme-linked immunosorbent assay (ELISA) in Indian patients with pemphigus vulgaris. AB - OBJECTIVE: To evaluate the role of the enzyme-linked immunosorbent assay (ELISA) test for the detection of antibodies to desmoglein 1 (dsg1) and desmoglein 3 (dsg3) in the diagnosis of pemphigus vulgaris (PV), and its correlation with disease severity and clinical presentation (mucosal PV, cutaneous PV, mucocutaneous PV). METHODS: Twenty-seven active PV patients and 26 controls with other dermatologic disorders were included in the study. The severity of oral and cutaneous involvement was assessed and recorded. ELISA test for the measurement of anti-dsg1 and anti-dsg3 antibodies was performed (Medical and Biological Laboratories Co. Ltd., Nagoya, Japan). The cut-off ELISA value for both anti-dsg1 and anti-dsg3 was taken as 20. RESULTS: Of the 27 patients, 26 were ELISA positive for anti-dsg1 antibodies and 23 for anti-dsg3 antibodies. Of the controls, two were positive for anti-dsg1 and none for anti-dsg3 antibodies. The sensitivity and specificity of ELISA for anti-dsg1 in the diagnosis of PV were 96.3% and 92.3%, respectively. For anti-dsg3, they were 85.2% and 100%, respectively. The different morphologic types of PV could not be differentiated on the basis of antibody profile; however, a direct correlation between anti-dsg3 titers and the severity of oral disease was noted, and also between anti-dsg1 titers and the severity of cutaneous disease. CONCLUSIONS: ELISA (dsg1 and dsg3) is an efficient tool for confirming the diagnosis of PV. Specific antibody titers correlate with disease severity; however, desmoglein testing cannot differentiate between the various morphologic subtypes of PV. PMID- 16700784 TI - Pemphigus in the Mediterranean region of Turkey: a study of 148 cases. AB - BACKGROUND: The purpose of this study was to describe the epidemiological and clinical features, course, response to treatment, and prognosis of pemphigus in the Mediterranean region of Turkey. METHODS: All patients with confirmed pemphigus were prospectively enrolled in two major dermatology departments in the cities of Adana and Antalya in the Mediterranean region between March 1998 and March 2004. Details including demography, findings of clinical examinations, treatment, course, and prognosis were recorded. RESULTS: One hundred and forty eight patients with pemphigus were diagnosed during the 6-year period, with a prevalence of 1.46 and an annual incidence of 0.24 per 100,000 in this region. There was a female predominance with a male to female ratio of 1:1.4. Pemphigus vulgaris (PV) was the most common clinical subtype, identified in 123 patients (83%). The mean age of onset was 43. In 101 (82%) patients with PV, disease began as persistent oral ulcers. The majority of the patients with PV could be managed with middle or high-dose steroids (60-140 mg/day). Complete clinical remission was obtained in 41 (39.4%) patients. The mortality rate was 4.8%. CONCLUSIONS: A moderately high incidence of pemphigus was found in the Mediterranean region of Turkey as compared with that encountered in other countries. The commonest clinical subtype was PV with a 9.5-fold higher incidence than pemphigus foliaceus. It is more frequent in middle-aged people and has a female predominance. Although a relatively higher dose of steroid was needed to control the PV, the disease completely remitted in a significant proportion of the patients. PMID- 16700785 TI - The association between malignant melanoma and noncutaneous malignancies. AB - BACKGROUND: Both increases and decreases in the incidence of subsequent malignancies in melanoma patients have been reported. We examined the database of the Indiana University Cancer Center to determine whether there is an association between malignant melanoma and noncutaneous malignancies. OBJECTIVE: We searched for evidence of noncutaneous malignancies in a cohort of melanoma patients. METHODS: Patients with microscopically confirmed malignant melanoma diagnosed between January 1987 and March 2001 were analyzed. This cohort was investigated for noncutaneous malignancies occurring either before or after the diagnosis of melanoma. The standardized incidence ratios (SIR) were calculated as the ratio of the observed to the expected number of patients with second malignancies, and 95% confidence intervals (95% CI) around the SIR were estimated from the cumulative Poisson distribution. RESULTS: A total of 955 patients with melanoma (498 males and 457 females) were documented over the 14-year period. Sixty-nine noncutaneous malignancies were identified in 59 (6.2%) melanoma patients (39 males and 20 females). There was a higher risk of non-Hodgkin's lymphoma (SIR = 1.91; 95% CI, 0.88-3.62) in men and renal cell carcinoma (SIR = 2.41, 95% CI, 0.97-4.97) in men. In female patients, however, there was no higher risk of noncutaneous malignancies. CONCLUSIONS: This study did not show a higher risk of prostate cancer, gastrointestinal cancer, leukemia, endometrial cancer, or cancer of the nerve and neuroendocrine systems in melanoma patients. No female patients incurred a higher risk of noncutaneous cancers. The increased risk of non Hodgkin's lymphoma and renal cell carcinoma in men might be attributed to a mutual carcinogenic exposure, an aberration of cell-mediated immunity, a shared genetic susceptibility, increased medical surveillance among cancer patients, a post-therapy effect after cancer management, or factors not as yet clear. Close monitoring of melanoma patients for signs of second malignancy is warranted. PMID- 16700786 TI - Attitudes of Iranian patients with skin problems towards using the Internet as a medical resource. AB - BACKGROUND: Internet use has grown rapidly in Iran, but there is no information about how Iranian Internet users gather medical information through this medium. OBJECTIVE: To study the use of the Internet as a medical resource by Iranian patients with chronic dermatologic problems. METHODS: We carried out a structured interview of all patients 20-40 years of age attending a private dermatology clinic between July and September 2003 for chronic skin disease. RESULTS: Of the 205 patients enrolled in the study, 104 (50.8%) had computer access and 74 (36.1% of the total) had Internet access. Nine patients (4.4%) had performed a search on the Internet for their skin condition, but only one had used an academic website. CONCLUSION: Despite the significant growth in Internet use in Iran, it appears that the Internet is not a significant medical resource for patients in Iran. PMID- 16700787 TI - Antibodies against desmoglein 1 in healthy subjects in endemic and nonendemic areas of pemphigus foliaceus (fogo selvagem) in Peru. AB - BACKGROUND: Endemic pemphigus foliaceus or fogo selvagem is an autoimmune skin disease characterized by the presence of subcorneal superficial blisters and antibodies of the immunoglobulin G4 (IgG4) class specific for the desmosomal glycoprotein, desmoglein 1. In Peru, no studies have been published on the seroprevalence of antibodies against desmoglein 1 in healthy subjects from endemic foci. SUBJECTS AND METHODS: This was a cross-sectional study. The sample included 82 healthy subjects, 41 from the Pueblo Libre community, a focus of endemic pemphigus foliaceus, and 41 from a nonendemic urban area in Pucallpa City. Enzyme-linked immunosorbent assay (ELISA) was used to determine the presence of antibodies against desmoglein 1. Samples were processed and tested at the Department of Dermatology and Cutaneous Surgery, School of Medicine, University of Miami, Miami, Florida. RESULTS: It was found that 31.7% of healthy individuals (13 subjects) from the endemic focus had anti-desmoglein 1 antibodies. A statistically significant association was found between the distance from the endemic focus and the presence of antibodies against desmoglein 1 in subjects living within the endemic focus [Mantel-Haenszel odds ratio (OR), 3.34; P = 0.03; 95% confidence interval (CI), 1.06-10.48]. Agriculture as an occupation showed a statistically significant association with the presence of antibodies against desmoglein 1 (Mantel-Haenszel OR, 7.84; P < 0.001; 95% CI, 2.47-24.87). CONCLUSIONS: Antibodies against desmoglein 1 are present in healthy subjects exposed to an endemic focus of pemphigus foliaceus (fogo selvagem). Agriculture is associated with a high risk of development of antibodies against desmoglein 1 in the endemic focus of the Pueblo Libre community. PMID- 16700788 TI - Quantification of blood intake of the head louse: Pediculus humanus capitis. AB - Although head lice, Pediculus humanus capitis, are globally prevalent blood sucking ectoparasites, the amount of blood imbibed by head lice has not been determined. This study investigated this parameter, as regular loss of a small quantity of blood may lead to an iron deficiency and anaemia. Adult female lice (66), adult males (46), and nymphs (152) were weighed before and after feeding in groups of 17-109 lice. The average amounts of blood imbibed at a single feed were: adult female louse (0.0001579 ml), adult male (0.0000657 ml) and nymph (0.0000387 ml). Assuming three feeds per day by an average infection of 30 lice (10 females, 10 males, and 10 nymphs), the average child with active pediculosis would loose 0.008 ml of blood per day. This amount of blood loss is of no clinical significance even in iron-deficient children. The most heavily infected child observed with 2657 lice could be expected to loose 0.7 ml/day or 20.8 ml/month, which may be of clinical importance in a child on an adequate diet, and would be significant in an iron-deficient child. However, if head lice feed more often than three times a day, a heavy infestation would have a greater potential to lead to iron deficiency. The frequency of feeding of head lice on the head of the human host needs to be determined. PMID- 16700789 TI - Peripheral blood mononuclear cells proliferation and Th1/Th2 cytokine production in response to streptococcal M protein in psoriatic patients. AB - BACKGROUND: Psoriasis is a chronic skin disease that is probably a T cell mediated autoimmune condition which is strongly associated with streptococcal throat infections. Although some groups have associated the involved response with different streptococcal antigens, M protein has been described as the major virulence factor of Streptococcus pyogenes. Thus, it is necessary to describe some features of the cellular responses to this streptococcal antigen. METHODS: Proliferation and Th1/Th2 cytokine production of peripheral blood mononuclear cells (PBMC) in response to total soluble extracts from type M5 S. pyogenes with (TSE37Sp) and without (M(-)TSESp) M protein were analyzed in 10 psoriatic patients and 10 healthy controls. RESULTS: PBMC from both patients and controls proliferated to both extracts. Responses to M(-)TSESp were significantly lower than those to TSE37Sp (P < 0.05). PBMC IL-2 and gammaIFN production after TSE37Sp stimulus was much higher than after M(-)TSESp antigenic stimulation in both groups (P < 0.05). Meanwhile, IL-4 production was quite low in both groups and in response to both extracts. We found a differential production of IL-10 between groups. PBMC from healthy controls responded to TSE37Sp with a much higher production of this cytokine as compared to the responses showed to M(-)TSESp while the cells from psoriatic patients responded without differences in the production of IL-10. CONCLUSION: Results obtained suggest an important Th1 response to M protein in psoriatic patients which could be associated with the cellular responses involved in psoriasis, while healthy subjects respond in a probably non-Th2 IL-10 producing regulatory T cells fashion. PMID- 16700790 TI - The frequency of scarring after genital ulcers in Behcet's syndrome: a prospective study. AB - BACKGROUND: Although genital ulceration with or without scar formation is an important manifestation of Behcet's syndrome, formal data on the rate of genital scarring are lacking. OBJECTIVE: To study prospectively the frequency of genital scar formation after genital ulcers in patients with Behcet's syndrome. Methods One hundred and two patients (72 males and 30 females) with fresh genital ulcers presenting at a dedicated Behcet's syndrome outpatient clinic were followed for 6 weeks. The ulcer location, size, and frequency of scar formation were noted. All patients were treated with similar topical therapy. RESULTS: Two hundred and ten genital ulcers were followed in both sexes. The rate of scar formation was 66.2% (102/154) in male patients and 60.7% (34/56) in female patients. When genital ulcers were stratified as a function of size, large genital ulcers (> or = 1 cm in diameter) developed scars more frequently than did smaller ulcers (< 1 cm). The rate of scar formation in male patients with large genital ulcers was 89.4% (59/66) and with small ulcers was 48.9% (43/88) (P < 0.001). In females, all 19 large ulcers healed with scarring, and the same was true for 40.5% (15/37) of the small ulcers (chi(2) = 18.61, P < 0.001). CONCLUSION About two-thirds of genital ulcers in Behcet's syndrome leave scars. The rate of scar formation depends on the ulcer size. PMID- 16700791 TI - Isolated eyebrow and eyelash trichotillomania mimicking alopecia areata. AB - BACKGROUND: Isolated Trichotillomania (TM) of the eyebrows and eyelashes is occasionally seen and may be mistaken for alopecia areata (AA). OBJECTIVE: The clinical study of the patients with isolated eyebrows and eyelashes alopecia suggesting of TM. PATIENTS AND METHOD: Seven children and young adult patients who referred with recalcitrant eyebrows and eyelashes alopecia were diagnosed as TM based on the clinical criteria for TM. The hair specifications for TM were include: hairs are broken at different levels, they may be tufted, tortuous and some hair fibers may be abnormally longer than others. The hair follicles may be prominent. The patients and their parents studied for psychopathology using a semistructured interview based on the Diagnostic Statistical manual of Psychiatric disorders (DSM-1V) and maudsley inventory. RESULTS: Our clinical study, interview with the patients and their parents and the clinical criteria used to differentiate TM from AA all support the clinical diagnosis of TM. The patients and their parents mainly suffered from obsessive-compulsive disorders (OCD) and obsessive-compulsive spectrum disorders (OCSDs). CONCLUSION: TM as a hair pulling habit in patients with underlying psychopathology can be considered in cases with isolated eyebrows and eyelashes alopecia when the hair are broken at different levels. And they may be malaligned, nonuniform, tortuous, and tufted. PMID- 16700792 TI - Characteristics of basal cell carcinoma amongst Asians in Singapore and a comparison between completely and incompletely excised tumors. AB - BACKGROUND: Most published series of basal cell carcinomas (BCCs) are based on Caucasian populations. Very little is known about the characteristics of incompletely excised BCCs in Asians. OBJECTIVES: This study examines the characteristics of BCCs in Asian residents in Singapore, and compares incompletely with completely excised tumors after conventional surgical excision. PATIENTS AND METHODS: A retrospective study was conducted using the case records of patients who had a primary BCC excised at the National Skin Centre between 1991 and 1995. RESULTS: A total of 185 histologically confirmed BCCs were recorded in 166 patients (male to female ratio, 0.95; mean age, 70.9 years). The highest proportion (27.1%) was in the 81-90-year age group. Compared with census data, a significantly higher proportion of patients with BCCs were Chinese (P < 0.001). The tumors averaged 12.0 mm in diameter and the majority (84.3%) were located on the head and neck. In 28 lesions (15.1%), the tumors were incompletely excised, with those on the nose and nasolabial folds being most frequent. Incompletely excised BCCs were more likely than completely excised lesions to be located on the mid-face and trunk (P = 0.003), but there was no significant correlation with tumor size, tumor duration, or patient age, race, and gender. The overall recurrence rate was 1.6% over a mean follow-up time of 74 months. CONCLUSIONS: BCCs are more common amongst Chinese in Singapore and occur chiefly in the elderly. Conventional surgical excision with margin control achieves a satisfactory tumor clearance rate of 84.9%. Incomplete excision is associated with tumor location on the mid-face and trunk rather than tumor size or duration. PMID- 16700793 TI - Androgenetic alopecia: does its presence change our perceptions? AB - BACKGROUND: The aim of this study was to evaluate the effects of androgenetic alopecia on males with and without hair loss and to delineate the level of stress gained by this type of alopecia. METHODS: Two hundred and 52 males (175 with hair loss, 77 without hair loss), between 16 and 72 years of age, participated in the study. A hair loss form (five questions for sociodemographical features, eight questions for dermatological features, eight questions for psychological evaluation) and a list of stressful life events were used. RESULTS: Desire for treatment was significantly different between the participants with and without androgenetic alopesia (AGA; chi(2) = 5.877, d.f. = 1, P = 0.015). Regardless of the presence of AGA, 61.4% accepted AGA as a natural condition with a solution that should be sought, 38.5% as a cause for increased attention, and 56.2% thought that the psychological effects of AGA would mostly be negative. The negative effects of their AGA on other family members, relationships with the opposite sex and occupation/academic life were considered by 37.1%, 43.0% and 36.3% of the participants, respectively. None of the psychological parameters differed significantly between the participants with and without AGA. A small portion (5%) of the variance in perception of stress (0.22(2) = 0.048) might be explained by the degree of alopecia. CONCLUSION: Social and cultural differences might alter perceptions concerning AGA regardless of its presence in an individual. PMID- 16700794 TI - Comparison of conventional and polymerase chain reaction diagnostic techniques for leishmaniasis in the endemic region of Adana, Turkey. AB - BACKGROUND: Cutaneous leishmaniasis (CL) has long been reported in the Cukurova region. We have compared the sensitivity of the conventional methods of diagnosis by microscopy and cultivation of lesion aspirates against polymerase chain reaction (PCR) amplification of parasite-specific DNA from these samples. METHODS: The samples (n = 25) were obtained from patients clinically diagnosed with CL at the regional dermatology clinic. Aliquots of the samples were stained with Giemsa for microscopy and cultured in Novy-Nicolle-McNeal (NNN) blood agar for promastigote growth. The remainder were subjected to DNA extraction for PCR amplification of the conserved region of kinetoplast minicircle DNA. PCR products of the expected size (120 bp) were observed after agarose gel electrophoresis, followed by staining with ethidium bromide. RESULTS: The positive rates from 25 samples were 44%, 68%, and 100% for cultivation, microscopy, and PCR, respectively. CONCLUSIONS: The PCR method used appears to be the most sensitive for the diagnosis of CL in this region. PMID- 16700795 TI - Chronic disseminated cutaneous histoplasmosis in an immunocompetent individual--a case report. PMID- 16700796 TI - Surgical pearl: tissue forceps as a simple and effective instrument for treating skin tags. AB - Skin tags are one of the most common, benign, disfiguring tumors on the flexural aspects of the body of advanced aged and obese people, with higher incidences in females. Various treatment modalities have been advocated for the tags. Even a simple, noninvasive and fast cryotechnique requires approximately 1-3 min for a single skin tag. To hasten the treatment of the tags, tissue forceps have been used as an effective, fast, simple instrument. By utilizing the tissue forceps, 37 skin tags at different parts of body have been treated successfully without cosmetic defects or recurrences at the sites. PMID- 16700797 TI - An unusual presentation of two simultaneous primary melanomas. PMID- 16700798 TI - Vitiligo: auto-immunity and immune responses. PMID- 16700799 TI - A case of cutaneous blastic NK lymphoma in a 32-year-old Korean woman. PMID- 16700800 TI - Case of multiple myeloma associated with extramedullary cutaneous plasmacytoma and pyoderma gangrenosum. AB - A variety of cutaneous manifestations has been described in multiple myeloma including extramedullary cutaneous plasmacytomas, cutaneous amyloidosis, pyoderma gangrenosum, leukocytoclastic vasculitis, necrobiotic xanthogranuloma, scleromyxedema, Sweet's syndrome, subcorneal pustular dermatosis, scleredema, and plane xanthomas etc. An 89-year-old Korean man, who had been suffering from multiple myeloma 1 year previous, presented for evaluation of two nodules on the right side of the forehead, left side of the chest (7th rib area), and multiple ulcers with papulopustules on both the thigh and the left side of the chest (2nd rib area) during 15 days, which developed at the same time. A biopsy of a lesion which manifested as a cutaneous nodule on the right side of the forehead revealed dermal infiltration by well-differentiated plasma cells, similar to those found on a bone marrow biopsy, and a biopsy of the lesion manifested as a painful ulceration on the right thigh area showing dermal neutrophilic infiltration. Histologic findings were consistent with plasmacytoma and pyoderma gangrenosum, respectively. We present a case of multiple myeloma which developed extramedullary cutaneous plasmacytoma and pyoderma gangrenosum simultaneously, which is very rare. The patient was treated with a systemic steroid and conservative therapy. PMID- 16700801 TI - Late-onset eccrine angiomatous hamartoma on the forehead. AB - A 71-year-old Korean man presented with a solitary erythematous plaque on his forehead (Fig. 1). It was first noticed by the patient 1 year previously and had slowly increased in size over that time. Physical examination revealed a slightly elevated, 1.5 cm x 1.5 cm erythematous plaque on the upper midline of the forehead. Sweating was not evoked by physical work or emotional stress. There was no pain or tenderness associated with the lesion. The patient had no history of trauma to the forehead. Histopathologic examination of the lesion showed increased numbers of eccrine glands, as well as dilated vascular channels in the deep dermis and subcutaneous tissue (Fig. 2). An immunohistochemical study showed that these eccrine glands stained positively for S-100 and carcinoembryonic antigen (CEA), and the vascular channels for the antifactor VIII-related antigens. These findings are consistent with eccrine angiomatous hamartoma. There was no change in the lesion during the 1-year follow-up period. PMID- 16700802 TI - Eberconazole 1% cream is an effective and safe alternative for dermatophytosis treatment: multicenter, randomized, double-blind, comparative trial with miconazole 2% cream. AB - BACKGROUND: Eberconazole is a topical, broad-spectrum imidazole derivative, effective in dermatophytoses, candidiasis, and pityriasis treatment. In previous trials, it showed a higher efficacy than clotrimazole in the treatment of dermatophytoses. The purpose of this trial was to evaluate the efficacy of eberconazole 1% cream compared with miconazole 2% cream in the treatment of dermatophytoses. METHODS: A multicenter, double-blind, randomized trial was performed in 653 patients with dermatophytoses, randomized to eberconazole 1% cream every 12 h or miconazole 2% cream every 12 h for 4 weeks. Treatment efficacy was assessed on the basis of the percentage of effective response after 4 weeks through mycologic and clinical assessment. RESULTS: Of the 653 patients included in the trial, 360 produced positive baseline mycologic cultures and were included in the efficacy assessment. Clinical efficacy was shown in 76.1% of patients receiving eberconazole and in 75.0% of patients receiving miconazole. The incidence of adverse events related to treatment was 0.91% for eberconazole and 0.92% for miconazole, none being serious, and all being local and transient. CONCLUSIONS: Eberconazole 1% cream is an effective treatment for fungal infections produced by dermatophytes, with a good safety and tolerability profile, and can be considered a good alternative for the treatment of dermatophytoses. PMID- 16700803 TI - Safety of efalizumab in adults with chronic moderate to severe plaque psoriasis: a phase IIIb, randomized, controlled trial. AB - BACKGROUND: To provide safety data for efalizumab, a recombinant humanized monoclonal IgG(1) antibody, in adults with chronic plaque psoriasis. METHODS: A 12-week, Phase IIIb, randomized, double-blind, parallel-group, placebo-controlled trial. At 58 study sites in the USA and Canada, 686 patients with moderate to severe chronic plaque psoriasis received an initial conditioning dose of efalizumab 0.7 mg/kg subcutaneously (SC) followed by either 11 weekly doses of efalizumab 1 mg/kg SC or matching placebo. Main outcome measures were safety and tolerability outcomes (primary) and efficacy outcomes (secondary). RESULTS: During 12 weeks of therapy with efalizumab or placebo, the incidence of clinical adverse events was 82.2% and 72.9%, respectively; the incidence of serious adverse events was 1.8% and 3.4%, respectively; and the incidence of nonserious adverse events leading to withdrawal was 1.8% and 1.7%, respectively. In the efalizumab group, there were no clinically significant changes in vital signs or laboratory parameters and no evidence of end-organ toxicities. A significantly higher proportion of patients receiving efalizumab than those receiving placebo achieved > or = 75% improvement in the Psoriasis Area and Severity Index (PASI) (P < 0.001), > or = 50% improvement in PASI (P < 0.001), and a static Physician's Global Assessment rating of Minimal or Clear (P < 0.001). The mean improvement in the Psoriasis Symptom Assessment was significantly greater in the efalizumab group (P < 0.001). CONCLUSIONS: Efalizumab treatment SC for 12 weeks was safe, well tolerated, and effective in patients with moderate to severe chronic plaque psoriasis. PMID- 16700804 TI - Two cases of generalized lichen nitidus treated successfully with narrow-band UV B phototherapy. AB - BACKGROUND: Narrow-band ultraviolet (UV) phototherapy has not been used in the treatment of lichen nitidus. AIM: To report two cases of generalized lichen nitidus successfully treated with narrow-band UV-B phototherapy. METHODS: A 7 year-old girl presented with a persisting, asymptomatic, papular eruption refractory to topical steroid for 3 months. Another 10-year-old boy presented with an asymptomatic papular eruption present for 6 months. The histopathologies of both lesions were consistent with lichen nitidus. The lesions were treated with narrow-band UV-B phototherapy. RESULTS: The lesions of the patients were almost completely cleared after the 30th and the 17th irradiation, respectively. CONCLUSIONS: Narrow-band UV-B may be an effective alternative therapy for the treatment of generalized lichen nitidus unresponsive to common therapies. PMID- 16700805 TI - Cutaneous findings in mummies from the British Museum. PMID- 16700808 TI - Kaposi's sarcoma-like angiosarcomas may reflect a common lymphatic endothelium differentiation pattern as Kaposi's sarcoma in association with chronic lymphedema. PMID- 16700810 TI - Flagellate erythema. PMID- 16700809 TI - Combined immunotherapy of recalcitrant warts. PMID- 16700811 TI - Epstein-Barr virus (EBV)-associated aggressive peripheral T-cell lymphoma showing folliculotropism. PMID- 16700812 TI - In reference to "Itching stretch marks and bullous lesions in a pregnant woman" by Uwe Wollina, Karl-Werner Degen, Helga Konrad, and Jacqueline Schonlebe. PMID- 16700813 TI - Airway selectivity: an update of pharmacokinetic factors affecting local and systemic disposition of inhaled steroids. AB - Topical corticosteroids remain the most efficacious single treatment for asthma and rhinitis, despite the emergence of newer drugs in recent years. The antiinflammatory properties of these products, combined with the targeting of formulations and optimization of the intrinsic pharmacokinetic features of the newer corticosteroid molecules has resulted in substantially improved airway selectivity. This review sets out to summarize the pharmacokinetic properties of inhaled corticosteroids that are important for the achievement of high levels of airway selectivity, with additional focus on the use of prodrugs/softdrugs relative to those of conventional corticosteroid molecules, mechanisms (such as esterification) by which retention at the target site is achieved while minimizing systemic exposure, and the role of plasma protein binding. PMID- 16700814 TI - Application of genomics in preclinical drug safety evaluation. AB - Understanding the response of biological systems to xenobiotics is fundamental to the evaluation of drug safety. Toxicologists have traditionally gathered pathological, morphological, chemical and biochemical information from in vivo studies of preclinical species in order to assess drug safety and to determine how new drugs can be safely administered to the human patient population. In recent years the emerging "-omics" technologies have been developed and integrated into preclinical studies in order to better assess drug safety by gaining information on the cellular and molecular events underlying adverse drug reactions. Genomics approaches in particular have become readily available and are being applied in several stages of drug development. The burgeoning literature on what has become known as "toxicogenomics" has for the most part highlighted successful applications of gene expression profiling in predictive toxicology, enabling decisions to be made on the developability of a compound early in the drug development process. It is also becoming apparent that toxicogenomic approaches are good starting points to develop experiments designed to gain a mechanistic insight into drug toxicities within and across species. Gene expression arrays permit the measurement of responses of essentially all the genes in the entire genome to be monitored, and knowledge of the function of the genes affected can identify the potential mechanisms to then be confirmed using conventional biochemical, toxicological and pathological approaches. As toxicologists put these technologies into practice they build up a knowledge base to better characterize toxicities at the molecular level and to make the search for much needed, novel biomarkers of toxicity more achievable. PMID- 16700816 TI - Effect of rifampicin on the pharmacokinetics of atenolol. AB - Also poorly metabolized drugs, including certain beta-blocking agents, can be susceptible to drug interactions caused by transporter inhibitors and inducers. Thus, our aim was to investigate the effect of rifampicin on the pharmacokinetics of atenolol in healthy people. In a randomized cross-over study with two phases, nine healthy volunteers received a 5-day pretreatment with rifampicin (600 mg daily) or placebo. On day 6, a single 100 mg dose of atenolol was administered orally. The plasma concentrations of atenolol and its excretion into urine were measured up to 33 hr after dosing. Systolic and diastolic blood pressures and heart rate were recorded in a sitting position before the intake of atenolol and 2, 4, 6, and 10 hr later. During the rifampicin phase, the mean area under the plasma concentration-time curve (AUC(0-infinity)) of atenolol was decreased to 81% and renal clearance increased to 109% of the placebo phase values (P<0.05). Rifampicin pretreatment reduced, albeit not statistically significantly, also the peak plasma concentration (Cmax), AUC(0-33 hr), and amount of atenolol excreted to 85% (P=0.139), 81% (P=0.053), and 86% (P=0.12) of the respective placebo phase values. The average heart rate and diastolic blood pressure were slightly higher during the rifampicin phase compared with the placebo phase (P<0.05). To conclude, although the inducing effect of rifampicin may not have been at its maximum by day 6, rifampicin has only a minor effect on the pharmacokinetics of atenolol evidenced by a slight reduction in its bioavailability. PMID- 16700815 TI - Pharmacodynamics of a new ophthalmic mydriatic insert in healthy volunteers: potential alternative as drug delivery system prior to cataract surgery. AB - Cataract surgery requires a satisfactory degree of mydriasis throughout the entire operation. A phase I, open-labelled, randomised, cross-over trial was conducted in 18 healthy volunteers to compare mydriasis obtained with subsequent administration of phenylephrine 10% and tropicamide 0.5% eyedrops or a new insoluble-matrix retropalpebral ophthalmic insert containing 5.38 mg phenylephrine and 0.28 mg tropicamide. Phenylephrine serum concentrations were measured over 6 hr following each treatment administration. Secondary end-points included cardiovascular, general and local tolerance and quantification of bacterial colonisation of the conjunctiva and the cultured insert, respectively. When normalized to the pupil diameter after conventional treatment, the diameter achieved with the insert was 1.13 (95% confidence interval, 0.94-1.48, P=0.38). Moreover, standard eye drops provided faster effective mydriasis than the insert, starting 30 min. as compared to 90 min. upon treatment administration (P<0.01, repeated-measures ANOVA). Phenylephrine concentrations remained almost undetectable for both treatments and no change in heart rate or blood pressure were observed throughout the study. Only three superficial punctuate keratitis were diagnosed with the insert and two with the eye drops. No significant bacterial contamination of conjunctiva swab and cultured insert was observed. The new insoluble-matrix retropalpebral ophthalmic mydriatic insert produced similar but delayed effective and prolonged mydriasis as compared to the standard delivery system. In addition to its potential usefulness in patients undergoing cataract surgery, such new ophthalmic delivery system may be an advantage in children who need to undergo fundus photography due to the single administration and excellent tolerance as well. PMID- 16700817 TI - Matrix metalloproteinase-9 activity in plasma correlates with plasma and whole blood lead concentrations. AB - Matrix metalloproteinases (MMPs) are enzymes involved in the degradation of the extracellular matrix. MMP-2 and MMP-9 have been implicated in a variety of pathological conditions including cardiovascular and neoplastic diseases, and recent studies have shown that circulating concentrations of MMP-9 may be a marker helping in the diagnosis and prognosis of cardiovascular and neoplastic diseases. We investigated whether there is an association between plasma MMP-2 and MMP-9 activities and the concentrations of lead in whole blood (blood Pb) or plasma (plasma Pb) from 40 lead-exposed persons (22 men and 18 women). Plasma Pb was determined by inductively coupled plasma mass spectrometry (ICP-MS) and blood Pb by graphite furnace atomic absorption spectrometry (GF-AAS). Plasma MMP-2 and MMP-9 activities were measured by gelatin zymography. We found a significant correlation between pro-MMP-9 activity in plasma and blood Pb (r=0.454; P=0.003), and between pro-MMP-9 activity in plasma and plasma Pb (r=0.312; P=0.049). No significant correlations were found between blood Pb or plasma Pb and plasma MMP 2. The association between pro-MMP-9 activity in plasma and both blood Pb and plasma Pb concentrations suggests a mechanism through which low lead exposure may increase the susceptibility to cardiovascular and neoplastic diseases. A causal relationship, however, remains to be proved. PMID- 16700818 TI - Effect of melatonin and atenolol on carbon monoxide cardiotoxicity: an experimental study in rats. AB - The aim of this study was to define the characteristics of heart rates and myocardial changes in rats exposed to carbon monoxide (CO), and the effects of reoxygenation, atenolol (a beta-blocker) and melatonin after sublethal CO intoxication. Widespread use of beta-blockers in cardiology practice and growing literature on the positive effect of melatonin in ischaemia reperfusion lead us to question their effects in case of CO intoxication. Rats were exposed to CO. After sublethal intoxication the rats were reoxygenated with ambient air. Subsequently blood values, electrocardiographic recordings and pathological changes were examined for each groups. Five rats died after CO intoxication in the control group: no myocardial changes were seen in light microscopy. However, myocardium of seven reoxygenated rats presented contraction bands. Seven reoxygenated rats pretreated with atenolol had a higher number of contraction bands of myocardial cells. Seven reoxygenated rats pretreated with melatonin had more contraction bands than reoxygenated rats, and heart rate recordings of these animals revealed a profund and sustained bradycardia. Thus, melatonin and atenolol appear to have some adverse effects in CO intoxication on the myocardial cells. PMID- 16700819 TI - Correlations between calcineurin phosphatase inhibition and cyclosporine metabolites concentrations in kidney transplant recipients: implications for immunoassays. AB - Cyclosporine exhibits a wide spectrum of metabolites that vary considerably in the extent to which they interfere with the various parent drug monitoring immunoassays. There is no consensus regarding the clinical significance of metabolites. Cyclosporine exerts its immunosuppressive action by inhibiting the enzyme calcineurin phosphatase. Determination of the enzyme's activity is one of the most promising pharmacodynamic markers. It is unknown how calcineurin phosphatase inhibition correlates with various cyclosporine monitoring assays and what is the potential impact of metabolites in this perspective? The aim of the present study was to determine the concentration of cyclosporine (by means of three different assay methods) and the four most significant metabolites (AM1, AM4N, AM9, and AM1C) in relation to calcineurin phosphatase inhibition. Twelve randomly selected cyclosporine-treated renal transplant patients were included in the study. Blood samples were drawn before, 1, 2, 3, 4, 6, 8, and 12 hr after oral intake of cyclosporine. Parent drug and metabolites were determined by liquid chromatography/tandem mass spectrometry (LC/MSMS). Additionally, cyclosporine concentration was determined by the enzyme multiplied immunoassay technique (EMIT) and by the polyclonal fluorescence polarization immunoassay (pFPIA). Calcineurin phosphatase activity was measured by its ability to dephosphorylate a previously phosphorylated 19-amino acid peptide. We found that calcineurin phosphatase inhibition correlates strongly with parent cyclosporine metabolites concentrations determined by all three assay methods. Determination methods that took metabolites into consideration exhibit stronger correlations with calcineurin phosphatase inhibition (sum of cyclosporin plus metabolites r= 0.93, LC/MSMS; pFPIA r=-0.94, PT in a Chinese and a Caucasian population. PMID- 16700827 TI - Subungual glomus tumor diagnosis based on imaging. AB - A 50-year-old woman had had tenderness of the nail bed of the right thumb for more than 20 years. For the previous 5 or 6 years, she had also had attacks of pain with exposure to cold, and deformity of the right thumb nail plate began to appear. There was red discoloration on the proximal aspect of the nail bed, and a longitudinal fissure on the distal aspect of the nail plate. Ultrasonography showed a well-circumscribed hypoechoic area under the proximal aspect of the nail plate and the nail matrix. Color Doppler ultrasonography showed subtle flow signals within the hypoechoic area. Magnetic resonance imaging showed a well circumscribed mass in the same place that the ultrasonography indicated. It was isointense to the dermis of the nail bed on the T1-weighted image and hyperintense on the T2-weighted image. Radiography showed subtle dorsal bone erosion in the distal phalanx. Surgery was performed. Histologically, the diagnosis of a subungual glomus tumor was made. We diagnosed the exact location and size of the subungual glomus tumor by preoperative imaging and completely removed it easily and safely. Imaging is very useful for diagnosing tumors of the nail unit. PMID- 16700828 TI - A case of perforating pilomatricoma. AB - Pilomatricoma is a rare skin neoplasm, most commonly seen in the head and neck region, and occurring in the first two decades of life. It is usually solitary and varies from 0.5 to 2 cm in diameter. Its etiology is unknown. Perforating pilomatricoma is a rare clinical variant that presents as a draining, crusted nodule or ulcer, and is reported to arise faster than the classic pilomatricoma. Herein, we report a case of 35-year-old female, who had a 4-month history of a growing mass on her leg. On physical examination, a 4-cm diameter, asymptomatic, erythematous, ulcerated mass was noted on the left anterio-lateral upper leg. The first histopathological analysis of a punch biopsy from the lesion was reported as basal cell carcinoma. Therefore, the lesion was totally excised. There were shadow cells, squamoid cells, and basaloid aggregations more prominently in the one area in the tumor. In addition, calcification, foreign body giant cells and inflammatory cells were present. Punch or excisional biopsies are preferred as a method of diagnosis for the majority of cutaneous neoplasms. If total excision is not the method of choice, multiple punch biopsies should be made from different areas in large skin tumors for correct diagnosis. PMID- 16700829 TI - Angiolymphoid hyperplasia with eosinophilia affecting the nail bed and underlying bone. AB - A 69-year-old Japanese woman had a dark red, 1-cm nodule located in the nail bed of the middle toe of her right foot. An X-ray examination revealed an osteolytic lesion of the distal phalanx of the right middle toe. Histopathology showed a proliferation of blood vessels surrounded by epithelioid cells and an infiltration of many eosinophils and some inflammatory cells. There has been no recurrence after surgical excision of the skin lesion and the distal phalanx of the right middle toe. PMID- 16700830 TI - Circumscribed palmar or plantar hypokeratosis: report of a Korean case and published work review. AB - Circumscribed palmar or plantar hypokeratosis is a rare dermatosis characterized by a solitary, well-circumscribed patch with scaly borders chiefly on the palm or sole; it usually occurs in middle-aged or elderly women. We report the case of a 52-year-old Korean woman with two characteristic lesions of circumscribed palmar hypokeratosis on the left palm. Clinically, the lesions simulated porokeratosis of Mibelli, but histologically there was no cornoid lamellation in the serial sections and there were the characteristic histopathological features of circumscribed palmar or plantar hypokeratosis including a stair-like configuration with an abrupt thinning of the stratum corneum and a decreased granular layer. We also review the 16 cases of circumscribed palmar or plantar hypokeratosis reported in the published work. PMID- 16700831 TI - Chromosome and sister chromatid exchange studies in Behcet's patients. AB - Behcet's disease is a chronic multisystemic disease of unknown pathogenesis characterized by four major symptoms: oral aphthous ulcers, skin lesions, ocular symptoms and genital ulcerations. The disease is spread throughout the world, but it is most frequent in Turkey, Japan, Korea and China. Although HLA-Bw51 has been found to predominate in Behcet's cases, the genetic etiology has not yet been clarified. In this study, we investigated the chromosomal abnormalities and sister chromatid exchange rates in patients with Behcet's diseases. Thirty-eight patients with Behcet's disease (diagnosed for the first time) and 30 healthy subjects (as controls) were included in this study. Although numerical and structural chromosomal abnormalities were not detected in our patients, we found an increased rate of sister chromatid exchange in patients over the control groups (P < 0.01). On the basis of these results, we discuss the genetic etiology of Behcet's disease. PMID- 16700832 TI - Melkersson-Rosenthal syndrome with diffuse facial swelling and multiple cranial nerve palsies. AB - We report an unusual case of Melkersson-Rosenthal syndrome presenting as diffuse facial swelling with facial, glossopharyngeal and vagus nerve palsies. A skin biopsy revealed non-caseating epithelioid cell granulomas throughout the dermis. The disease responded poorly to systemic steroids, antihistamines, antibiotics and potassium iodide. PMID- 16700833 TI - A mutilating arthropathy, "rhupus hands" associated with multiple synovial cysts in a patient with systemic lupus erythematosus. AB - "Rhupus hands" is a phrase coined to describe one of the deforming arthropathies associated with systemic lupus erythematosus (SLE), because the clinical features are indistinguishable from those of rheumatoid arthritis. Herein, we report a case of rhupus hands with multiple synovial cysts arising in a 60-year-old woman with SLE. PMID- 16700834 TI - A case of erythrokeratoderma variabilis: loosened gap junctions in the acanthotic epidermis. AB - A 15-year-old Japanese female without contributory personal or family medical history had demonstrated irregular, keratotic plaques in the lower extremities since infancy that had been gradually enlarging. The keratotic plaques showed partial erythematous change, which altered shape over a relatively short period, leaving pigmentation. The biopsy specimen taken from the erythematous, keratotic plaque showed typical church-spire-like papillomatosis with acanthosis, and thickening of granular and horny layers. Gene analysis targeting connexin 30.3 and 31, based on the diagnosis of erythrokeratoderma variabilis, did not demonstrate any abnormality of these genes. However, ultrastructural observation disclosed an increased amount of gap junctions, some of which showed four layers on high-powered electron microscopy, suggesting loosened connection of the plasma membrane of the keratinocytes through the gap junctions. This loosened gap junction structure was also observed in a case of lamellar ichthyosis, examined as a reference. The disturbed cell-to-cell interaction through latent damage to the gap junctions may be related to the keratotic changes of the epidermis in these skin diseases. PMID- 16700835 TI - Food-induced anaphylaxis caused by ingestion of royal jelly. AB - We report a case of food-induced anaphylaxis caused by ingestion of royal jelly. After taking royal jelly and several other medicinal products, a 33-year-old Japanese male developed severe facial pruritus and erythema, followed by vertigo, numbness in his fingers, generalized pruritus, wheals, dyspnea, wheezing and impaired consciousness. He was treated with corticosteroid and fluid therapy, and his symptoms subsided. Upon allergy testing, his only positive reaction was to royal jelly. Given the clinical symptoms and the positive prick test to royal jelly, a diagnosis of anaphylaxis due to the ingestion of royal jelly was made. In this paper, we describe this case and review the relevant published work. PMID- 16700836 TI - Fibrous hamartoma of infancy manifesting as multiple nodules with hypertrichosis. AB - Fibrous hamartoma of infancy is a rare benign tumor, which usually occurs within the first year of life. Most cases present as a solitary, painless, skin-colored subcutaneous nodule; multiple lesions are rarely reported. There have also been a few cases which have showed overlying skin changes, including hypertrichosis and alterations in pigmentation. We report a rare case of fibrous hamartoma of infancy in a 6-month-old female who presented as multiple nodules with overlying hypertrichosis on the left buttock. To the best of our knowledge, there have been only two reported cases in the dermatological published work that showed these unusual findings simultaneously. PMID- 16700837 TI - A case of peeling skin syndrome successfully treated with topical calcipotriol. PMID- 16700838 TI - Nevoid acanthosis nigricans localized to the umbilicus. PMID- 16700839 TI - Adams-Oliver syndrome: a case with complete expression. PMID- 16700840 TI - Pachyonychia congenita with only nail involvement. PMID- 16700841 TI - ASTEROID--when spinning a star leads to a collision with reality. PMID- 16700842 TI - Cardiology referrals--getting to the heart of the matter. PMID- 16700843 TI - The neurobiological basis for partial agonist treatment of nicotine dependence. PMID- 16700844 TI - Using item-specificity to promote tailored training and remediation in normal and pathological aging. PMID- 16700845 TI - A simple drawing test to identify patients who are unlikely to be able to learn to use an inhaler. AB - Previous research has shown that patients are not able to learn to use self administered inhaler devices if they have impaired general cognition, executive (frontal lobe) dysfunction or dyspraxia. These impairments are most frequently encountered in frail elderly people. Some of the tests required to detect such impairments are time-consuming and are therefore not easily adopted in routine clinical practice, particularly in an outpatient setting. We performed a study of 50 elderly inhaler-naive patients to explore the use of a simple quick drawing test (copying overlapping pentagons) to determine whether that test was able to identify patients who were unable to learn to use a Turbohaler. Patients who were not able to perform the pentagon copying test were found to be unable to learn to use a Turbohaler with a specificity of 93% and positive predictive value of 87%. This was at least as discriminating as the Abbreviated Mental Test and the Mini Mental State Examination. The overlapping pentagon drawing test is useful in clinical practice to identify patients who will probably not use a self administered inhaler correctly, and who will require an alternative approach to the treatment of their airways disease. This finding might have implications for other self-administered treatment, such as insulin pens and complex drug regimens. PMID- 16700846 TI - Frequency of Hoover's sign in stable patients with chronic obstructive pulmonary disease. AB - Hoover's sign (the inward motion of the lower lateral rib cage with inspiration) is conventionally considered to be a sign of severe disease in chronic obstructive pulmonary disease (COPD). However, no studies have been done regarding the frequency of Hoover's sign in patients with stable COPD. We aim to establish the frequency of Hoover's sign in a large series of stable patients with COPD and to analyse the characteristics associated with its presence. One hundred and fifty-seven consecutive patients with COPD, 150 of whom were men (95%), with a mean (standard deviation) age of 68 (8) years were included. Seventy-one patients had Hoover's sign (45%) on clinical examination. Hoover's sign was not detected in mild COPD patients, and it was present in 36% of moderate, 43% of severe and 76% of very severe COPD patients. In the multivariate analysis, dyspnea, body mass index (BMI), number of exacerbations and number of prescribed drugs were independently associated with the presence of Hoover's sign in COPD. Hoover's sign is a frequent finding in COPD, and the frequency increases with severity. Its presence is independently related to higher values of dyspnea, BMI, number of exacerbations and number of prescribed drugs. PMID- 16700847 TI - Low-molecular-weight heparin, bemiparin, in the outpatient treatment and secondary prophylaxis of venous thromboembolism in standard clinical practice: the ESFERA Study. AB - The objective of this study is to assess the clinical and economic outcomes associated with outpatient treatment and secondary prophylaxis of acute venous thromboembolism (VTE) with a low-molecular-weight heparin, bemiparin. This study was designed as an open-label, multicentre, prospective, cohort study in standard clinical practice. Sixty-three investigators from 54 Spanish centres participated in the study. Five hundred eighty-three patients (434 outpatients and 149 inpatients) with acute VTE were followed up for 98 days (median). Outcome measures were costs and adverse events during initial VTE treatment with bemiparin (outpatient vs. inpatient cohorts) and long-term treatment [bemiparin (BEM) vs. vitamin K antagonists (VKA) cohorts]. Mean total costs per patient were lower in the outpatient cohort as compared with those in the inpatient cohort (1206 vs. 5191 euros; difference = -3985 euros; p < 0.001), with similar rates of adverse events (5.1 outpatient vs. 7.4% inpatient; p = 0.196) over 98 days. Mean total costs per patient were similar in the BEM/BEM and BEM/VKA cohorts (3616 vs. 3831 euros; difference = -215 euros; p = 0.412), but patients on long-term bemiparin treatment had lower rates of major bleeding (0.4 vs. 1.7%; p = 0.047), minor bleeding (1.8 vs. 6%; p = 0.032) and total adverse events (2.9 vs. 9.5%; p = 0.007) than patients in the BEM/VKA cohort. Outpatient management of VTE with bemiparin in selected patients resulted in significant cost-savings compared to inpatient treatment, while maintaining effectiveness and safety. Bemiparin may be a safer and cost-neutral alternative to VKA for long-term treatment of VTE. PMID- 16700848 TI - Effect of sedation on soluble P-selectin levels, coagulation and myocardial damage following a coronary intervention. AB - This study was aimed to investigate the effect of sedation on plasma soluble P selectin (sP-selectin) levels and initiation phase of coagulation following coronary intervention. A total of 58 patients with unstable angina were randomised into two groups of sedated (n = 29) and nonsedated (n = 29) patients. sP-Selectin, adrenaline, noradrenaline, von Willebrand factor (vWF), fibrinogen, platelet and troponin-I levels were measured before the intervention and at 30 min and 24 h after the intervention. In the sedated group, adrenaline, noradrenaline, sP-selectin and fibrinogen levels remained stable 30 min after the intervention (p > 0.05), whereas vWF level increased (p < 0.05). In the nonsedated group, 30-min sP-selectin (p < 0.001), adrenaline (p < 0.05), noradrenaline (p = 0.012), vWF (p < 0.001) and fibrinogen (p < 0.001) levels were found to be increased. Also, 24-h sP-selectin, vWF and fibrinogen levels for both groups showed significant increases when compared with the values both 30 h before (p < 0.001) and 30 min after the intervention (p < 0.001). Sedation before interventions reduces sP-selectin levels, initial phase of coagulation and, consequently, myocardial damage. PMID- 16700849 TI - Efficacy study of galantamine in possible Alzheimer's disease with or without cerebrovascular disease and vascular dementia in Thai patients: a slow-titration regimen. AB - The objective is to evaluate the efficacy of galantamine when a slow titration regimen is employed in Thai Alzheimer's disease (AD) patients with or without cerebrovascular disease and vascular dementia (VaD). A 6-month, multicentre, open label, uncontrolled trial was undertaken in 75 AD patients. Eligible patients received an initial galantamine dose of 8 mg/day and escalated over 5-8 weeks to maintenance doses of 16 or 24 mg/day. Primary efficacy measures were AD Assessment Scale-cognitive subscale (ADAS-cog) and the Clinician's Interview Based Impression of Change-Plus version (CIBIC-plus). The Behavioural Pathology in AD Rating Scale (BEHAVE AD), the AD Cooperative Study Activities of Daily Living Inventory and Pittsburgh Sleep Quality Index were the secondary efficacy variables. Analyses were based on the intent-to-treat population. Treatment with galantamine showed significant improvement in cognition on the ADAS-cog and CIBIC plus at month 6. Galantamine showed favourable effects on activities of daily living. Behavioural symptoms and sleep quality were also significantly improved (p < 0.05). Galantamine was well tolerated. The adverse events were mild-to moderate intensity. The most frequent adverse events commonly reported were nausea (16.4%), dizziness (9.6%) and vomiting (6.8%). The results of this study may be consistent with galantamine being an effective and safe treatment for mild to-moderate AD patients with or without cerebrovascular disease and VaD. Flexible dose escalation of galantamine was well tolerated. The daily maintenance dose of galantamine was 16 mg/day, followed by a back up dose of 24 mg/day. PMID- 16700850 TI - Stratification of foot ulcer risk in patients with diabetes: a population-based study. AB - This trial assessed whether a simple clinical tool can be used to stratify patients with diabetes, according to risk of developing foot ulceration. This was a prospective, observational follow-up study of 3526 patients with diabetes (91% type 2 diabetes) attending for routine diabetes care. Mean age was 64.7 (range 15 101) years and duration of diabetes was 8.8 (+/-1.5 SD) years. Patients were categorised into 'low' (64%), 'moderate' (23%) or 'high' (13%) risk of developing foot ulcers by trained staff using five clinical criteria during routine patient care. During follow-up (1.7 years), 166 (4.7%) patients developed an ulcer. Foot ulceration was 83 times more common in high risk and six times more in moderate risk, compared with low-risk patients. The negative predictive value of a 'low risk score' was 99.6% (99.5-99.7%; 95% confidence interval). This clinical tool accurately predicted foot ulceration in routine practice and could be used direct scarce podiatry resources towards those at greatest need. PMID- 16700851 TI - NSAID-related upper gastrointestinal bleeding: are risk factors considered during prophylaxis? AB - The purposes of this study were to evaluate the effect of nonsteroidal anti inflammatory drugs (NSAIDs) on acute nonvaricose upper gastrointestinal bleeding (ANUGIB) and establish whether the NSAID-prescribing physicians take precautions to prevent or reduce GI ulcerations. Clinical characteristics, causes of bleeding and clinical outcomes of patients hospitalised in our gastroenterology clinic with ANUGIB were recorded prospectively over a 1.5-year period. NSAIDs, including aspirin, were used by 127 of 168 patients (73%). Among the NSAID users, 100 patients (78%) had at least one risk factor for serious adverse GI events related to NSAIDs. Only two patients were using proton pump inhibitors and one patient was using H2 receptor blocker of the high-risk group for GI side effects of NSAIDs. NSAIDs have an important effect on GI bleeding, and it seems that risk factors are underestimated by physicians. PMID- 16700852 TI - Tranquilliser use as a risk factor for falls in hospital patients. AB - This study looked at associations of tranquilliser use and falls risk in a hospital population of confused and nonconfused patients. In a prospective observational study in a rehabilitation hospital for elderly patients, we followed 1025 consecutive patients. The number of fallers, recurrent fallers and total falls was recorded. Confused patients (p < 0.0001) and patients on tranquillisers (p = 0.001) were significantly more likely to fall than nonconfused patients and patients off tranquillisers. Confused patients on tranquillisers were significantly more likely to have recurrent falls (p = 0.026) when compared with confused patients off tranquillisers. The risk was apparent from admission, persisting throughout the first 30 days of stay. This was not noted for nonconfused patients. We identified a stratification of risk for falls with nonsignificant trends for confused and nonconfused patients on tranquillisers to be fallers and to have more falls compared with patients off tranquillisers. These data are associational and do not necessarily imply causality. There is however no evidence to recommend the routine withdrawal of tranquillisers from all patients. Any future research needs to include confused patients. PMID- 16700854 TI - Diagnostic approach in patients with asymptomatic haematuria: efficient or not? AB - Many patients with asymptomatic haematuria are primarily referred to urological examination. To analyse the efficiency of this strategy, we investigated the diagnostic process in 134 patients with asymptomatic haematuria (91 macroscopic and 43 microscopic) who were referred to our urology department. The frequency of diagnostic procedures was urine culture: 91%, cystoscopy: 87%, intravenous urography: 86%, ultrasound: 73%, examination of urinary sediment: 20%. In 70 patients (52%), a urological diagnosis was made. In the remaining 64 patients, the urinary sediment was examined for the presence of dysmorphic erythrocytes and erythrocyte casts in 17 patients (27%). Signs of glomerular haematuria were found in nine of them (53%). The diagnostic strategy in these patients with asymptomatic haematuria was not discordant from several published algorithms but appeared neither efficacious nor efficient. We estimate that when examination of the urinary sediment would have been performed at the start of work-up, 25% of patients could have spared from extensive urological investigation. PMID- 16700853 TI - C-arm-guided pudendal nerve block: a new technique. AB - Pudendal nerve block (PNB) is an effective diagnostic and/or treatment method for perineal pain. Various approach techniques, such as transperineal, transvaginal, computerised tomography (CT)- or sono-guided approach, have been suggested for this block. However, they have some limitations, such as high cost, difficulty to perform in practice, inaccurate and unreliable results and inconvenience. To overcome these limitations, we first tried C-arm-guided approach for accomplishing PNB in the prone position. Under the optimal ischial spine view of C-arm fluoroscopy, the block needle was placed on the tip of the ischial spine. Then a mixed solution for the block was administered. All of the 25 patients enrolled in this study were blocked successfully using this method. No side effects or complications were observed in relation to the block. We concluded that the C-arm-guided approach for PNB is an effective alternative to the existing techniques, which can overcome their limitations. PMID- 16700855 TI - Ultracision reduces acute blood loss but not seroma formation after mastectomy and axillary dissection: a pilot study. AB - Various surgical cutting coagulation devices are used during breast surgery. A pilot study was set up to determine if ultracision would reduce complications of intraoperative blood loss and seroma formation. Acute blood loss was significantly reduced in the harmonic scalpel group in comparison with the control group. However, there was little difference between the two groups in terms of hospital stay, volume or duration of postoperative drainage or subsequent aspiration of seroma. PMID- 16700856 TI - Health care providers' perceptions of the problems and causes of irrational use of drugs in two Middle East countries. AB - It is now evident that both developed and developing countries are experiencing many aspects of inappropriate use of drugs in their health care facilities. This is the first study in the region performed to examine the most common problems of irrational use of drugs and their causes in two Middle East countries--Jordan and Syria. Ninety senior participants from Jordan (50-15 physicians and 35 pharmacists) and Syria (40-12 physicians and 28 pharmacists) were enrolled in this study. The participants were asked to fill two questionnaires that deal with the problems and causes of irrational use of drugs in their country. Additionally, the participants were asked to perform a prescription analysis using WHO prescribing indicators on 40 prescriptions taken randomly from a comprehensive health centre in their country. The main drug use problems identified in the two countries were almost the same, but they vary in the percentage of occurrence and include excessive use of antibiotics and antidiarrhoeals, overprescribing of nonsteroidal anti-inflammatory drugs, prescribing by tradename, excessive use of antibiotics to treat minor upper respiratory infections and self-medication by the public. The main causes of irrational use of drugs were poor medical records, lack of patient education about illnesses and drugs, no family doctor system, lack of standard treatment guidelines and lack of continuing medical education for doctors and pharmacists. The results of this study are important for decision-makers to utilise when putting policies and strategies to improve the use of drugs in both countries. PMID- 16700857 TI - The neurobiological basis for partial agonist treatment of nicotine dependence: varenicline. AB - Smoking cessation has major health benefits for men and women of all ages. However, most smokers are addicted to nicotine and fail repeatedly in their attempts to quit. Stimulation of nicotinic receptors in the brain, particularly alpha4beta2 receptors, releases dopamine in the meso-limbic area of the brain and is reinforcing. Nicotine abstinence reduces dopamine release, and this is associated with withdrawal symptoms and craving for nicotine. Eight current pharmacotherapies--bupropion, nortriptyline, clonidine and nicotine patch, gum, inhaler, lozenge and nasal spray--are moderately effective aids to smoking cessation. Each is significantly better than placebo, but approximately 80% of patients using one of these medications return to smoking within the first year. Varenicline, a specific alpha4beta2 nicotinic receptor partial agonist, is a new pharmacotherapy that stimulates dopamine and simultaneously blocks nicotine receptors. Phase II and III trials have yielded promising results suggesting that varenicline could be an important advance in the treatment of nicotine dependence. PMID- 16700858 TI - Importance of renin in blood pressure regulation and therapeutic potential of renin inhibition. AB - Despite improvements in its detection and treatment, hypertension remains a significant public health problem worldwide. In recent years, many international hypertension societies and organisations have set increasingly rigorous blood pressure (BP) targets, with the aim of reducing cardiovascular complications, and this has in turn necessitated the use of more antihypertensive medications to reach these targets in individual patients. There is therefore an ongoing need to develop antihypertensive drugs with new mechanisms of action. Renin inhibitors represent a novel class of compounds which offer considerable promise as BP lowering agents. Here, we review the rationale for renin inhibition as a therapeutic target and examine the preclinical and clinical evidence for the antihypertensive effectiveness of the renin inhibitors. PMID- 16700859 TI - A review of the management of patients after percutaneous coronary intervention. AB - The exponential increase in the numbers of percutaneous coronary interventions (PCIs) has led to many clinicians having to care for post-PCI patients. We review the management of early problems seen in post-PCI patients, such as vascular access site complications, contrast nephropathy, drug-induced thrombocytopaenia and chest pain. The management of possible restenosis and the use of stress testing are discussed. The complications from dual antiplatelet therapy are addressed. The prognosis of the post-PCI patient, the implications of co-existent heart failure and the newer technologies of implantable defibrillator and cardiac resynchronisation therapy are reviewed. We conclude by emphasising the importance of secondary prevention by risk factor modification as well as the communication between the clinician and the cardiologist. PMID- 16700860 TI - Diabetes prevention: is there more to it than lifestyle changes? AB - Over the past years, there has been an explosive increase in the prevalence of type 2 diabetes (T2DM) and this is expected to continue, entailing associated morbidity and mortality. An increasing number of studies explore the different ways T2DM could be prevented. On-going lifestyle modifications need to be addressed. High-risk patients should be given counselling on weight loss, possibly using a low glycaemic index diet, with a target of around 7-10% over 6 12 months, as well as instruction for increasing physical activity to around 150 min of physical exercise weekly (NNT = 4-8). Moderate alcohol consumption and coffee consumption may also be of benefit (NNT = 89 and 66, respectively). Metformin (NNT = 14), acarbose (NNT = 11) and troglitazone (NNT = 6) have been shown to prevent/delay T2DM and angiotensin-converting enzyme (ACE) inhibitors and statins appear to have an adjunctive role (NNT = 42 and 112, respectively). Trials with orlistat and bariatric surgery have also prevented T2DM (NNT = 36 and 6, respectively), and forthcoming treatment with GLP1 mimetics appears promising. Diabetes prevention studies should help create well-defined strategies for screening and treating high-risk populations in the real world, as prevention is our only chance to alleviate the ever growing burden of diabetes mellitus in the world. PMID- 16700861 TI - Contemporary referral of patients from community care to cardiology lack diagnostic and clinical detail. AB - The quantity of referrals to secondary care is increasing. That the quality of medical referrals is decreasing is a common allegation yet has rarely been assessed. We report a time-limited, cross-sectional survey evaluating cardiological referral information quality. Referral letters (n = 218, excluding direct access pro formas) from GPs to the Cardiology Department at City Hospital, Birmingham, were collated and analysed over 2 months. A subset (n = 49) of these patients completed questionnaires assessing their knowledge and patient communication of the referral. Information quality was poor (length, diagnosis, expectation, prior treatment and investigation) with almost half of all letters containing only outline symptomatic complaints without diagnosis. The majority of patients referred had not been investigated or treated in any way before referral. Despite lack of understanding of the reason for referral, typically the majority of patients expressed themselves as satisfied with the process. Given most referrals are seen as appropriate, information exchange between secondary and primary care is crucial. By contrast, the standard of even basic clinical assessment communicated between primary care and secondary care was severely limited. The reason(s) why medical assessment is lacking are unclear but must be explored to give more support to primary care to complete basic medical task particularly if investment is to flow into this source. PMID- 16700862 TI - Audit of bowel preparation with Picolax (sodium picosulfate plus magnesium citrate) for colonoscopy. AB - The aim of this study was to assess the quality of bowel preparation for colonoscopy with standard dose Picolax (two sachets administered on the day prior to the procedure, as per the manufacturer's instructions) in a large cohort of patients. A retrospective audit was performed of colonoscopies performed at our institution over a 1-year period. Patients were excluded if standard dose Picolax was not used, if the quality of the bowel preparation was not recorded or if completion of the procedure was not recorded. Of the 619 fully evaluable cases, the quality of the bowel preparation was assessed by the colonoscopist performing the procedure as good in 263 (42.5%), satisfactory in 242 (39.1%) and poor in 114 (18.4%) of the cases. In only 28 (4.5%) cases, poor bowel preparation was the reason cited for an incomplete colonoscopy. There was no difference in the quality of bowel preparation between inpatients and outpatients. In clinical practice, Picolax is an effective bowel preparation for colonoscopy in the vast majority of cases. PMID- 16700863 TI - Metoclopramide for migraine-associated hiccup. AB - Persistent hiccup can be a distressing disorder. I present a case of migraine attack-associated sustained hiccup. Metoclopramide can swiftly control both hiccup and migraine headache. Vasopressin release probably underlies migraine aborting action of metoclopramide while restoration of oesophageal smooth muscle function involves competitive dopaminergic antagonism and a prominent cholinergic agonist activity. Episodic prolonged hiccup associated with nausea is an unusual presenting feature of migraine. PMID- 16700864 TI - Torsades de pointes in congenital long QT syndrome following low-dose orphenadrine. AB - We report the case of a woman, affected by congenital long QT syndrome (LQTS), who experienced three syncopal episodes shortly after the assumption of a low dose of orphenadrine. The ECG revealed a QT interval of 600 ms, and the corrected QT interval (QTc) was 537 ms. No structural cardiac disease was demonstrated by echocardiography. Orphenadrine treatment was discontinued. During the first 12 h of monitoring, three short-lasting, asymptomatic episodes of torsades de pointes occurred. No other sustained ventricular arrhythmia was revealed at Holter monitoring in the following days. During the ensuing 6 months, the patient remained asymptomatic, and the QTc did not change. Orphenadrine is an analogue of diphenhydramine, an antihistaminic drug that produces sodium channel blockade similar to that caused by quinidine and other Class Ia antiarrhythmic drugs. Our case rises the suspicion that orphenadrine could cause life-threatening arrhythmias in LQTS even at a low dose, and independently from concomitant assumption of potentially QT-prolonging drugs. PMID- 16700865 TI - Response to: Appropriate choice of proton pump inhibitor therapy in the prevention and management of NSAID-related gastrointestinal damage. PMID- 16700867 TI - Response to: Are all non-thymidine analogue backbones appropriate for treating antiretroviral-naive patients? PMID- 16700869 TI - Duloxetine 60 mg once daily in the treatment of milder major depressive disorder. AB - There is ongoing debate regarding the effectiveness of antidepressants in patients with milder major depressive disorder (MDD). This post-hoc analysis evaluated the efficacy and tolerability of duloxetine in the subset of 159 (75 duloxetine and 84 placebo) patients with milder MDD (baseline HAMD17 total score > or = 15 and < or = 18) who were treated once daily with duloxetine 60 mg or placebo in two identical, 9-week, randomised, double-blind trials. At endpoint, change from baseline on HAMD17 was greater in the duloxetine group (-7.0) than in the placebo group (-4.1) (p = 0.005). Response and remission rates, and improvement on the Clinical Global Impressions-Severity (CGI-S) scale, the Patient Global Impressions-Improvement (PGI-I) scale, and measures of painful symptoms were also significantly better in the duloxetine group (p < 0.05). Tolerability was consistent with that seen in previous studies of duloxetine in patients with more severe depression. In conclusion, duloxetine 60 mg/day is effective and well tolerated in milder MDD. PMID- 16700870 TI - Selective imidazoline agonist moxonidine in obese hypertensive patients. AB - Obesity is the major risk factor for the development of hypertension. This association accentuates the risk of cardiovascular disease, as it is frequently accompanied by the components of the metabolic syndrome. This randomised open parallel study evaluated the chronic effects of moxonidine--a selective imidazoline receptor agonist--on blood pressure, plasma catecholamines, leptin, insulin and components of the metabolic syndrome in obese hypertensives. Amlodipine was used as the control drug. Our results showed that moxonidine and amlodipine significantly reduced blood pressure when measured using the oscillometric method and 24-hour blood pressure monitoring. Moxonidine therapy decreased systolic blood pressure from 160.4 +/- 2.4 to 142.1 +/- 3.3 mmHg (p < 0.005) and diastolic blood pressure from 102.4 +/- 1.3 to 89.7 +/- 1.6 mmHg (p < 0.005) after 24 weeks of treatment. Moxonidine administration reduced the supine arterial plasma levels of adrenaline from 63.2 +/- 6.6 to 49.0 +/- 6.7 pg/ml (p < 0.005), the supine arterial plasma levels of noradrenaline from 187.9 +/- 10.7 to 149.7 +/- 13.2 pg/ml (p < 0.01) and the orthostatic venous plasma levels of noradrenaline from 258.6 +/- 25.0 to 190.3 +/- 16.4 pg/ml (p = 0.03). Those variables were not changed by amlodipine. The plasma levels of leptin and insulin 120 min after a glucose load decreased after moxonidine administration from 27.2 +/- 3.5 to 22.6 +/- 2.9 pg/ml (p < 0.05) and from 139.7 +/- 31.2 to 76.0 +/- 15.2 U/ml (p < 0.05), respectively. Amlodipine, however, did not modify those variables. This study showed a comparable reduction in blood pressure with both antihypertensive drugs. Moxonidine decreased sympathetic nervous activity, improved insulin resistance and reduced the plasma levels of leptin. PMID- 16700872 TI - Impact of obesity and insulin resistance on vasomotor tone: nitric oxide and beyond. AB - 1. Obesity is rapidly increasing in Western populations, driving a parallel increase in hypertension, diabetes and vascular disease. Prior to the development of overt diabetes or hypertension, obese patients spend years in a state of progressive insulin resistance and metabolic disease. Mounting evidence suggests that this insulin-resistant state has deleterious effects on the control of blood flow, thus placing organ systems at a higher risk for end-organ damage and increasing cardiovascular mortality. 2. The purpose of the present review is to examine the current literature on the effects of obesity and insulin resistance on the acute control of vascular tone. Effects on nitric oxide (NO)-mediated control of vascular tone are particularly examined with regard to proximal causes and distal mechanisms of the impaired NO-mediation of vasodilation. 3. Finally, novel pathways of impaired control of perfusion are summarized from the recent literature to identify new avenues of exploring impaired vascular function in patients with metabolic disease. PMID- 16700873 TI - Cytidine 5'-diphosphocholine restores blood flow of superior mesenteric and renal arteries and prolongs survival time in haemorrhaged anaesthetized rats. AB - 1. The aim of the present study was to investigate the effect of the intracerebroventricular (i.c.v.) or intravenous (i.v.) administration of cytidine 5 cent-diphosphocholine (CDP-choline) on superior mesenteric artery (SMA) and renal artery (RA) blood flow, along with the cardiovascular parameters and survival time of anaesthetized rats under conditions of haemorrhagic shock. 2. Rats were anaesthetized with urethane (1.25 g/kg, i.p.) and acute haemorrhage was mimicked by the withdrawal of a total volume of 2-2.1 mL blood/100 g bodyweight over a period of 20 min. The CDP-choline was injected i.c.v. (1.0, 1.5 and 2.0 micromol) or i.v. (250 mg/kg) after the end of haemorrhage. Blood pressure, heart rate, SMA and RA flow values and the survival time of rats were recorded. Changes in blood flow were estimated by laser-Doppler flowmetry. 3. The haemorrhage procedure decreased the blood pressures of rats by 60% and limited their survival time to 22 +/- 2 min. Both SMA and RA flow decreased to approximately 25% of initial values at the end of the haemorrhage procedure. 4. The i.c.v. administration of CDP-choline (1.0, 1.5 and 2.0 mmol) increased blood pressure and partially reversed the hypotension in a dose- and time-dependent manner. At 1.5 and 2.0 mmol, i.c.v., CDP-choline completely restored the decreased flow of the RA and transiently reversed hypoperfusion of the SMA. It also produced an almost fourfold increase in the survival time of rats. 5. The i.v. administration of CDP-choline (250 mg/kg) also completely, but transiently, restored SMA and RA flow, whereas it increased blood pressure by only 40% compared with control values. The survival time of rats in the i.v. CDP-choline group was doubled that of control. 6. These results indicate that both centrally and peripherally injected CDP-choline can restore SMA and RA flow, together with a partial reversal of hypotension and an increase in the survival time of rats. PMID- 16700874 TI - Blockade of beta1- and beta2-adrenoceptors delays wound contraction and re epithelialization in rats. AB - 1. The participation of sympathetic efferent fibres in wound healing is not well understood. The aim of the present study was to investigate the effects of beta(1)- and beta(2)-adrenoceptor blockade on rat excisional cutaneous wound healing. 2. Male rats were treated orally with propranolol dissolved in drinking water (50 mg/kg per day), whereas the control group received drinking water without propranolol. Propranolol was administered daily until rats were killed. A full-thickness excisional lesion was performed. The lesion area was measured to evaluate wound contraction. After rats had been killed, lesion and adjacent normal skin were formol fixed and paraffin embedded. Sections were stained with haematoxylin-eosin, Sirius red or Toluidine blue and immunostained for a-smooth muscle actin or proliferating cell nuclear antigen. 3. Propranolol-treated rats presented delayed wound contraction and epidermal healing and decreased hydroxyproline levels, collagen density and neo-epidermis thickness. Blockade of beta(1)- and beta(2)-adrenoceptors increased epidermal and connective tissue cell proliferation, polymorphonuclear leucocyte migration, myofibroblast density and mast cell migration. The volume density of blood vessels was increased and vessels were more dilated in propranolol-treated animals. 4. Thus, we conclude that beta(1)- and beta(2)-adrenoceptor blockade impairs cutaneous wound healing. This information should be considered by physicians during the treatment of patients who present with hypertension and problems in the healing process (such as venous ulcers). PMID- 16700875 TI - Effects of autonomic blockade on non-linear cardiovascular variability indices in rats. AB - 1. The present study assesses the effects of autonomic blockade (alpha- and beta adrenoceptor and cholinergic) on cardiovascular function studied by heart rate variability (HRV), blood pressure variability (BPV) and baroreflex sensitivity in rats using non-linear dynamics. Little is known about the influence of pharmacological autonomic nervous system interventions on non-linear cardiovascular regulatory indices. 2. In 13 conscious rats, heart rate and aortic blood pressure were measured continuously before, during and after autonomic blockade with atropine, phentolamine and propranolol. Non-linear scaling properties were studied using 1/f slope, fractal dimension and long- and short term correlation. Non-linear complexity was described with correlation dimension, Lyapunov exponent and approximate entropy. Non-linear indices were compared with linear time and frequency domain indices. 3. Beta-adrenoceptor blockade did not alter the non-linear characteristics of HRV and BPV, although low-frequency power of HRV was depressed. Alpha-adrenoceptor blockade decreased the scaling behaviour of HRV, whereas cholinergic blockade decreased the complexity of the non-linear system of HRV. For BPV, the scaling behaviour was increased during alpha adrenoceptor blockade and the complexity was increased during cholinergic blockade. The linear indices of HRV and BPV were decreased. 4. The present results indicate that the beta-adrenoceptor system has little involvement in the generation of non-linear HRV and BPV in rats. 5. Alpha-adrenoceptor blockade mostly influenced the scaling properties of the time series, whereas cholinergic blockade induced changes in the complexity measures. 6. The absence of the baroreflex mechanism can trigger a compensatory feed-forward system increasing the complexity of BPV. PMID- 16700876 TI - Hyperinsulinaemia increases the gene expression of endothelial nitric oxide synthase and the phosphatidylinositol 3-kinase/Akt pathway in rat aorta. AB - 1. Hyperinsulinaemia has been reported to be an independent risk factor for cardiovascular diseases. Insulin stimulates both the phosphatidylinositol 3 kinase (PI3-K)/Akt and mitogen-activated protein kinase (MAPK) pathways. To investigate the direct effects of insulin on vascular tissues, we examined the gene and protein expression of insulin signalling molecules, endothelial nitric oxide synthase (eNOS) and MAPK in aortas obtained from established hyperinsulinaemic rats under deep urethane anaesthesia (1.2 g/kg, i.p.). 2. High plasma insulin levels significantly enhanced the gene and protein expression of eNOS in aortas. This was accompanied not only by increased mRNA levels of insulin receptor substrate (IRS)-1, IRS-2, PI3-K and Akt, but also by a high protein content of Akt and phospho-Akt (Ser473). 3. In contrast, MAPK mRNA levels were decreased in hyperinsulinaemic rats compared with normoinsulinaemic rats. 4. Insulin receptor mRNA levels were also lower in insulin-treated rats rather than controls. The overexpression of mRNA for vascular endothelial growth factor (VEGF) and insulin-like growth factor (IGF)-I receptor was also observed in aortas from hyperinsulinaemic rats. 5. To our knowledge, these data provide the first direct measurements of the mRNA of insulin signalling molecules and the downstream eNOS and MAPK. We conclude that hyperinsulinaemia itself can lead to the upregulation of eNOS and the PI3-K/Akt pathway in the vasculature and may also induce the overexpression of VEGF and IGF-I receptor genes. PMID- 16700877 TI - Vasorelaxing effects of propranolol in rat aorta and mesenteric artery: a role for nitric oxide and calcium entry blockade. AB - 1. Propranolol has been prescribed successfully to patients with cardiovascular diseases, but the exact mechanisms by which it reduces peripheral vascular resistance have been poorly investigated. 2. The present study was designed to investigate the relaxing effects of propranolol in the rat isolated aorta and mesenteric artery, focusing on the contribution of the nitric oxide (NO)-cGMP pathway and calcium entry blockade. Relaxation responses to propranolol were obtained in precontracted rat aortic and mesenteric artery rings. 3. DL Propranolol (10-100 micromol/L) produced concentration-dependent relaxations in the aorta and mesenteric artery rings with intact endothelium. The isomers D- and L-propranolol produced relaxation responses that were equipotent to the racemic mixture. 4. Metoprolol (10-100 micromol/L) produced slight relaxations, whereas atenolol (10-100 micromol/L) had no relaxant activity. 5. The NO inhibitor N(G) nitro-L-arginine methyl ester (100 micromol/L) and the soluble guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (1 micromol/L), as well as removal of the endothelium, significantly reduced the relaxation responses induced by the lower concentrations of propranolol without affecting maximal responses. In addition, DL-propranolol markedly increased cGMP levels in endothelium-intact preparations. 6. In Ca(2+)-free Krebs' solution, DL propranolol (10-100 micromol/L) caused marked rightward shift in the concentration-response curves to CaCl(2), with a decrease of maximal responses in tissues with either intact or denuded endothelium. Nifedipine (1 micromol/L) in combination with DL-propranolol virtually abolished the CaCl(2)-induced contractile responses. 7. The relaxation responses induced by DL-propranolol were significantly reduced in aortic and mesenteric rings precontracted with phorbol 12,13-dibutyrate (1 micromol/L). 8. In conclusion, DL-propranolol relaxes arterial smooth muscle by mechanisms involving activation of the NO-cGMP pathway and calcium influx blockade, independent of beta-adrenoceptor blockade. PMID- 16700878 TI - Rats with inherited stress-induced arterial hypertension (ISIAH strain) display specific quantitative trait loci for blood pressure and for body and kidney weight on chromosome 1. AB - 1. The aim of the present study was to scan chromosome 1 in the hypertensive 'inherited stress-induced arterial hypertension' (ISIAH) rat strain for the quantitative trait loci (QTL) that control basal and stress-induced arterial blood pressure (ABP) levels and weight traits. 2. Two F(2) populations of 3-4- and 6-month-old male rats derived from a cross between the normotensive Wistar albino Glaxo (WAG) and hypertensive ISIAH rats were used in the search for the QTL. To identify the QTL for blood pressure (basal and under stress) and weight traits (bodyweight, as well as the weight of the adrenals, kidney and heart), 12 polymorphic markers covering a span of 234.6 Mb on chromosome 1 were analysed. 3. In 3-4-month-old rats, QTL were found for bodyweight in the vicinity of the D1Rat76 marker (230.6 Mb; P = 0.0019; logarithm of odds (LOD) score 3.23) and for relative kidney weight in the vicinity of the D1Rat117 marker (219.3 Mb; P = 0.000992; LOD score 3.41). No QTL for blood pressure were detected on chromosome 1 in the 3-4-month-old population. 4. In 6-month-old rats, a QTL for basal ABP in the region spanning 168.0-250.4 Mb, with two peaks around the markers D1Rat168 (204.8 Mb; P = 0.00087; LOD score 3.42) and D1Rat76 (P = 0.0006; LOD score 3.34), was described. A novel QTL was found in the D1Rat54-D1Rat168 region for stress induced blood pressure (P = 0.0014; LOD score 3.08). 5. The results provide support for the existence of age-dependent differences in the genetic control of ABP and weight traits. Chromosome 1 was characterized by four QTL: for bodyweight and relative kidney weight in 3-4-month-old F(2) (ISIAH yen WAG) rats and basal ABP and ABP under emotional (restraint) stress conditions in 6-month-old F(2) rats. The QTL for stress-induced ABP seems to be novel and specific to the ISIAH rat strain. PMID- 16700879 TI - Effect of theophylline and aminophylline on transmitter release at the mammalian neuromuscular junction is not mediated by cAMP. AB - 1. Theophylline and aminophylline have been widely used as inhibitors of phosphodiesterase when examining the role of cAMP in regulating cell function. In reality, however, these phosphodiesterase inhibitors may have additional sites of action that could complicate the interpretation of the results. These additional sites of action could include antagonism of inhibitory adenosine autoreceptors and release of intracellular calcium. The purpose of the present study was to determine which of the above three is the primary mechanism by which theophylline and aminophylline affect transmitter release at the mammalian neuromuscular junction. 2. Quantal release measurements were made using intracellular recording techniques. A variety of drugs were used to elucidate this pathway. Isoproterenol, an adenylate cyclase activator, was first used to establish the effect of enhanced levels of cAMP. Theophylline application on its own or in the presence of a drug combination that blocked the adenosine receptor and phosphodiesterase pathways caused significant release depression, opposite to what is expected if it was functioning to enhance cAMP levels. However, when applied in the presence of a drug combination that blocked the adenosine receptor, phosphodiesterase and intracellular ryanodine calcium pathways, theophylline was unable to depress release. Therefore, it was concluded that the major mechanism of action of theophylline is depression of transmitter release by causing the release of intracellular calcium. 3. Aminophylline application alone resulted in a significant enhancement of release. However, when coupled with an adenosine receptor blocker, the ability of aminophylline to enhance transmitter release was blocked, suggesting that its dominant mechanism of action is adenosine receptor inhibition. 4. Taken together, these results indicate that the use of theophylline and aminophylline is inappropriate when examining the role of cAMP at the mammalian neuromuscular junction. PMID- 16700880 TI - Dissociation of blood pressure and sympathetic activation of renin release in sinoaortic-denervated rats. AB - 1. Blood pressure (BP) and heart rate (HR) increase 6 and 24 h after sinoaortic baroreceptor denervation (SAD), whereas plasma renin activity (PRA) and renal renin mRNA levels remain unchanged. We postulated that a simultaneous rise in BP could offset the expected activation of renin associated with an increased renal sympathetic discharge secondary to SAD. 2. To test this hypothesis, the increase in BP associated with the onset of SAD was prevented by a continuous infusion of sodium nitroprusside (SNP; 30 microg/kg per h). Changes were measured in five groups of conscious adult male Wistar rats: (i) sham; (ii) SAD; (iii) SAD rats in which the BP was prevented from increasing by infusion of SNP; (iv) sham rats in which the BP was increased by 30% by infusion of phenylephrine (PE; 1.5-2.0 mL/h); and (v) SNP + PE for 3 h by infusion as above. 3. As expected, BP and heart rate (HR) increased significantly following SAD compared with sham rats (152 +/- 4 vs 116 +/- 3 mmHg, respectively, for BP and 503 +/- 6 vs 345 +/- 13 b.p.m., respectively for HR; n = 5; P < 0.05) but remained unchanged when SNP was infused for 3 h (106 +/- 1 mmHg and 455 +/- 9 b.p.m., respectively; n = 5; P < 0.05). 4. Similarly, BP and HR increased with PE infusion compared with PE + SNP (138 +/- 9.9 vs 113 +/- 2.3 mmHg for BP, respectively, and 325 +/- 9 vs 423 +/- 18 b.p.m. for HR, respectively; n = 5; P < 0.05). 5. Plasma renin activity remained unchanged in SAD compared with sham rats (1.67 +/- 0.35 vs 1.05 +/- 0.17 ng angiotensin (Ang) I/mL per h), but increased significantly when hypertension was prevented (5.86 +/- 0.77 ng AngI/mL per h; n = 5; P < 0.05). Renin mRNA levels in the kidneys were unchanged in all groups. 6. These results show that an elevation in BP appears to offset increased renal sympathetic discharge with no change in PRA. PMID- 16700881 TI - Spironolactone further reduces urinary albumin excretion and plasma B-type natriuretic peptide levels in hypertensive type II diabetes treated with angiotensin-converting enzyme inhibitor. AB - 1. Over the course of treatment with angiotensin-converting enzyme inhibitor (ACEI), plasma levels of aldosterone have been shown to increase and this increase would blunt the effectiveness of the ACEI (aldosterone escape phenomenon). 2. In the present study, we assessed a potential renal benefit of additional aldosterone blockade with spironolactone in hypertensive diabetic patients treated with ACEI showing the phase of aldosterone escape. 3. The present clinical study was a randomized prospective study to assess difference between the clinical effects of spironolactone and furosemide. Thirty hypertensive type II diabetics (DM2) with a urinary alubumin:creatinine ratio (ACR) above 30 mg/g creatinine (showing albuminuria) and plasma B-type natriuretic peptide (BNP) levels above 100 pg/mL (showing mild heart failure) were treated with an ACEI (imidapril 5 mg/day) for 1 year and then randomly divided into two groups, one group receiving additional spironolactone (25 mg/day) treatment and the other receiving furosemide (20 mg/day) treatment. Blood pressure, ACR and plasma BNP levels were monitored in both groups. 4. Treatment with the ACEI reduced ACR initially but, in 1 year, ACR tended to increase. Additional spironolactone treatment progressively reduced ACR, whereas furosemide treatment did not show any effect. Plasma BNP levels were reduced by ACEI and were further reduced by additional spironolactone treatment, but not furosemide treatment. Blood pressure levels in both groups were comparable. 5. In conclusion, additional therapy with spironolactone in ACEI treatment exerts a renoprotective, as well as cardioprotective, effect in hypertensive diabetes. PMID- 16700883 TI - Monoclonal antibodies as targeting and therapeutic agents: prospects for liver transplantation, hepatitis and hepatocellular carcinoma. AB - 1. Monoclonal antibodies (mAbs) of high specificity and stability have become key resources in the therapeutic, diagnostic and drug discovery fields to treat various immunological disorders and malignancies of different organs. 2. The latest genetic engineering technology applied in antibody design and production, such as phage display technology and genetically modified mouse, have revolutionized the clinical applicability and feasibility of the use of mAbs in humans. 3. Innovative antibody products in the forms of single-chain or super humanized antibody therapeutics having a higher affinity for target antigens and minimal antigenicity in hosts have been introduced for experimental purposes and/or clinical trials. 4. Although there are successful examples of antibody therapeutics in the market, the use of mAbs in treating hepatitis-related disease and hepatocellular carcinoma is rare and remains to be exploited. PMID- 16700884 TI - Human embryonic stem cells: technological challenges towards therapy. AB - 1. Human embryonic stem cells (hESC) hold promise for overcoming many diseases because they provide a potential source for many of the slow-growing cell types needed for effective tissue repair, such as the dopaminergic neural cells for Parkinson's disease or the pancreatic islet cells needed to relieve diabetic patients of their daily insulin injections. 2. Human embryonic stem cells can be characterized by several surface antigen markers, transcription factors and enzymes, as well as their ability to differentiate into cells representative of the three germ layers, both in vivo and in vitro. 3. Significant progress has been made in defining the feeder-free and serum-free conditions needed for the culture of hESC. The fibroblast growth factor-2 and transforming growth factor-b signalling pathways appear to be important in maintaining self-renewal and preventing differentiation, respectively. 4. Several important quality controls, including karyotyping, immunogenicity and murine viral assays, will have to be established to monitor the production of hESC for therapeutic purposes. 5. Methods of expansion and differentiation of hESC are still in their infancy and the efficiency of these processes needs to be significantly enhanced. PMID- 16700885 TI - DNA microarray technology for target identification and validation. AB - 1. Microarrays, a recent development, provide a revolutionary platform to analyse thousands of genes at once. They have enormous potential in the study of biological processes in health and disease and, perhaps, microarrays have become crucial tools in diagnostic applications and drug discovery. 2. Microarray based studies have provided the essential impetus for biomedical experiments, such as identification of disease-causing genes in malignancies and regulatory genes in the cell cycle mechanism. Microarrays can identify genes for new and unique potential drug targets, predict drug responsiveness for individual patients and, finally, initiate gene therapy and prevention strategies. 3. The present article reviews the principles and technological concerns, as well as the steps involved in obtaining and analysing of data. Furthermore, applications of microarray based experiments in drug target identifications and validation strategies are discussed. 4. To exemplify how this tool can be useful, in the present review we provide an overview of some of the past and potential future aspects of microarray technology and present a broad overview of this rapidly growing field. PMID- 16700887 TI - Filarial nematode secreted product ES-62 is an anti-inflammatory agent: therapeutic potential of small molecule derivatives and ES-62 peptide mimetics. AB - 1. The 'hygiene hypothesis' postulates that the recent increased incidence of allergic or autoimmune diseases (e.g. asthma, type I diabetes) in the West reflects an absence of appropriate priming of the immune response by infectious agents, such as parasitic worms, during childhood. 2. Consistent with this, it has long been recognized that several autoimmune disorders, such as rheumatoid arthritis (RA), a T helper (Th) 1-mediated autoimmune disease characterized by excess production of pro-inflammatory cytokines, such as tumour necrosis factor alpha, exhibit reduced incidence and severity in geographical regions with high parasite load, suggesting that environmental factors may subtly alter disease progression. 3. Infection with worms also appears to suppress Th2-biased inflammatory disorders, such as asthma, because there also appears to be an inverse correlation between parasite load and atopy. This is perhaps more surprising, given that helminths often induce strong Th2-type immune responses characterized by release of specific cytokines, such as interleukin (IL)-4, IL-5 and IL-13. 4. Therefore, these findings suggest that the co-evolution of helminths with hosts, which has resulted in the ability of worms to modulate inflammatory responses in order to promote parasite survival, may also have generated a predisposition for the host to develop autoimmunity and allergy in the absence of infection. 5. The mechanisms underlying such immunomodulation are not clear, but appear to involve the release of parasite-derived molecules that allow the worms to modulate or evade the host immune response by a number of mechanisms, including skewing of cytokine responses and the induction of T regulatory cells. 6. In the present review we discuss the properties of one such filarial nematode-derived immunomodulatory molecule, namely ES-62, its anti inflammatory action and the therapeutic potential of small molecule derivatives and peptides that mimic its action. PMID- 16700886 TI - Short interfering RNA (siRNA) as a novel therapeutic. AB - 1. RNA interference (RNAi) is a robust method of post-transcriptional silencing of genes using double-stranded RNA (dsRNA) with sequence homology driven specificity. The dsRNA can be between 21 and 23 nucleotides long: this is converted to small interfering RNA (siRNA), which then mediates gene silencing by degradation/blocking of translation of the target mRNA. 2. RNA interference provides a simple, fast and cost-effective alternative to existing gene targeting approaches both in vitro and in vivo. The discovery of siRNAs that cause RNAi in mammalian cells opened the door to the therapeutic use of siRNAs. Highly intense research efforts are now aimed at developing siRNAs for therapeutic purposes. 3. Recent advances in the design and delivery of targeting molecules now allow efficient and highly specific gene silencing in mammalian systems. Synthetic siRNA libraries targeting thousands of mammalian genes are publicly available for high-throughput genetic screens for target discovery and validation. Recent studies have demonstrated the clinical potential of aptly designed siRNAs in various types of viral infections, cancer and renal and neurodegenerative disorders. 4. The present review provides insight into the novel therapeutic strategies of siRNA technology, which is the latest development in nucleic acid based tools for knocking down gene expression, and its potential for silencing genes associated with various human diseases. PMID- 16700888 TI - CD4+CD25+ regulatory T cells in health and disease. AB - 1. Over the past 5 years, tremendous progress has been made in understanding the suppressive mechanisms of T regulatory (Treg) cells. The Treg cells, a subpopulation of T cells, have been shown to play an important role in maintaining peripheral tolerance and the prevention of autoimmunity. 2. Various populations of Treg cells have been described, including thymically derived CD4(+)CD25(+) Treg cells. These naturally occurring Treg cells are present in the periphery and are capable of suppressing proliferation and effector T cell responses both in vitro and in vivo. 3. In addition, a second subset of Treg cells, type 1 T regulatoary (Tr1) and Th3 cells, exert their suppressive capacity via cytokines such as interleukin-10 and transforming growth factor-beta and are contact independent. 4. The present review summarizes the characteristics and molecular basis of CD4(+)CD25(+) Treg cells, as well as their therapeutic potential in modulating inflammatory diseases, such as inflammatory bowel disease and rheumatoid arthritis. PMID- 16700889 TI - Pharmacogenetics approach to therapeutics. AB - 1. Pharmacogenetics refers to the study of genetically controlled variations in drug response. Functional variants caused by single nucleotide polymorphisms (SNPs) in genes encoding drug-metabolising enzymes, transporters, ion channels and drug receptors have been known to be associated with interindividual and interethnic variation in drug response. Genetic variations in these genes play a role in influencing the efficacy and toxicity of medications. 2. Rapid, precise and cost-effective high-throughput technological platforms are essential for performing large-scale mutational analysis of genetic markers involved in the aetiology of variable responses to drug therapy. 3. The application of a pharmacogenetics approach to therapeutics in general clinical practice is still far from being achieved today owing to various constraints, such as limited accessibility of technology, inadequate knowledge, ambiguity of the role of variants and ethical concerns. 4. Drug actions are determined by the interplay of several genes encoding different proteins involved in various biochemical pathways. With rapidly emerging SNP discovery technological platforms and widespread knowledge on the role of SNPs in disease susceptibility and variability in drug response, the pharmacogenetics approach to therapeutics is anticipated to take off in the not-too-distant future. This will present profound clinical, economic and social implications for health care. PMID- 16700891 TI - Therapeutic vaccination for central nervous system repair. AB - 1. Vaccination against infectious agents has been heralded as a triumph in modern medicine and, more recently, cancer vaccines have risen in prominence. The present review looks towards the use of vaccine therapy to attenuate damage after injury to the central nervous system (CNS). 2. Significant debility is associated with brain injury, most commonly occurring as a result of physical trauma or stroke. This end result reflects the inability of neurons and axons to regenerate following injury to the CNS. This unconductive environment is due, in large part, to the presence of myelin and oligodendrocyte-related inhibitors of neurite outgrowth. 3. We review how a vaccine-based approach has been variably used to circumvent this issue and promote axonal regeneration and repair following traumatic injury and other neurodegenerative disorders. In addition, emerging evidence suggests that the immune response to injury in the CNS may be manipulated so as to reduce cellular damage. Vaccine-directed approaches using this concept are also outlined. PMID- 16700890 TI - Antisense oligonucleotides: from design to therapeutic application. AB - 1. An antisense oligonucleotide (ASO) is a short strand of deoxyribonucleotide analogue that hybridizes with the complementary mRNA in a sequence-specific manner via Watson-Crick base pairing. Formation of the ASO-mRNA heteroduplex either triggers RNase H activity, leading to mRNA degradation, induces translational arrest by steric hindrance of ribosomal activity, interferes with mRNA maturation by inhibiting splicing or destabilizes pre-mRNA in the nucleus, resulting in downregulation of target protein expression. 2. The ASO is not only a useful experimental tool in protein target identification and validation, but also a highly selective therapeutic strategy for diseases with dysregulated protein expression. 3. In the present review, we discuss various theoretical approaches to rational design of ASO, chemical modifications of ASO, ASO delivery systems and ASO-related toxicology. Finally, we survey ASO drugs in various current clinical studies. PMID- 16700892 TI - DNA vaccines and allergic diseases. AB - 1. Allergic diseases are characterized by inappropriate immune responses to common environmental antigens. The prevalence of these diseases has been increasing worldwide for reasons that are not exactly clear. 2. Current treatment is largely symptomatic. Because the initial observation that simple plasmid DNA injections resulted in in vivo protein expression and induction of adaptive immune responses to the encoded antigen, the potential of modifying the allergic immune responses by DNA vaccination so as to treat and prevent these diseases has been explored extensively. 3. In the present paper we review preclinical studies using animal models of allergic diseases, with an emphasis on DNA vaccine design, for house dust mite allergens-related allergic asthma. PMID- 16700893 TI - Art and science of photodynamic therapy. AB - 1. Photodynamic therapy is an established modality for the treatment of solid tumours and other accessible lesions. Although the concept and practice of combining light with a photosensitizing agent for the treatment of disease states has been around for almost a century, the understanding of the art and science therein has been tremendously enhanced over the past few years. 2. Photosensitized reactions are dependent on the generation of reactive oxygen species, in particular singlet oxygen, which accounts for the damaging effects on biological macromolecules, such as membrane lipids and proteins. Therefore, compounds that give a good yield of (1)O(2) are used as photosensitizers. 3. The main photosensitizers used in the clinical setting belong to the photofrin family; however, newer and more effective sensitizers are being evaluated for their potential clinical effectiveness. 4. Light sources have moved from the use of white light with specific filters in the old days to the more recent use of monochromatic light sources, such as lasers, to more sophisticated light-emitting diodes. However, dosimetry remains a big issue mainly because of difficulties in establishing the optimum treatment conditions for an approach that requires the fine-tuning of several variables, such as sensitizer and light doses and drug-to light interval, as well as the issues of skin photosensitivity and low selectivity. A newer development to circumvent these and provide a broader application for this concept has been the phenomenon of photo-activation, whereby photo-exposure of chromophores to generate novel, small biologically active compounds has been demonstrated successfully. 5. The aim of the present review was to provide a general overview of the art and science of photodynamic therapy and to highlight some of the issues and recent developments in further advancing this modality of treatment. PMID- 16700895 TI - Proteomics technology and therapeutics. AB - 1. Proteins are the workhorses of biological systems and, thus, it is not surprising that the field of proteomics has flourished in recent years. The present review briefly surveys the general proteomic approaches taken to identify drug targets and to evaluate drug efficacy and toxicity in the preclinical and clinical settings. These generally fall into three basic categories: a profiling approach, a functional approach and a structural approach. 2. The current 'tools' used and applied to the drug-discovery process will be highlighted. These include the two-dimensional gel electrophoresis, liquid chromatography, mass spectrometry, isotope-coded affinity tag and protein biochips. 3. Clearly, proteomics has begun to set a foothold in every stage of the drug-discovery process. Future developments in this area will likely make a significant impact in our quest for better, safer and cost-effective drugs. PMID- 16700894 TI - Polymeric core-shell nanoparticles for therapeutics. AB - 1. Nanobiotechnologies have recently attracted significant attention from chemists, biologists, engineers and pharmaceutical scientists. In particular, they have been widely applied to improve drug, protein/peptide and gene delivery. 2. This review presents recent advances in the field of drug, protein/peptide and gene delivery using natural and synthetic polymer nanoparticles and explains how polymeric nanoparticles are specifically designed to suit the needs for targeted delivery of small molecular drugs, proteins/peptides and genes. In addition, some of the challenges and prospects for these technologies are discussed. PMID- 16700896 TI - Targeting tumours by adoptive transfer of immune cells. AB - 1. Surgery, radiotherapy and chemotherapy are the most widely used and well established modalities for treating malignant diseases. Surgery is used to excise solid tumours and radiotherapy/chemotherapy are used for the treatment of liquid tumours and for solid tumours where there is a risk of micrometastases. A major drawback for both radiotherapy and chemotherapy is their lack of specificity for tumour cells. Both these treatments can destroy normal bone marrow cells and result in severe side-effects. 2. The impairment of haemapoiesis due to bone marrow destruction combined with the use of toxins in chemotherapy that inhibit the proliferation of immune cells leaves many patients immunocompromised. This complicates the development of prophylactic (vaccine) strategies for tumours where patients are undergoing conventional therapy. 3. An alternative approach is to expand and activate tumour-specific immune cells in vitro that can then be adoptively transferred back in large numbers. This is defined as adoptive immunotherapy and has the advantage of potentially bypassing the immuno inhibitory effects of conventional therapies. 4. Transferred immune cells have been shown to mediate tumour regression in patients by both direct and indirect mechanisms. The immune cells used include tumour reactive T lymphocytes and dendritic cells, which elicit tumour specific responses. 5. Many novel cell-based immunotherapeutic strategies developed in murine tumour models are now being applied in human clinical trials. The malignancies targeted include melanoma, chronic myelogenous leukaemia and breast, ovarian, colon and kidney cancers. In the present review, we discuss these novel cell-based strategies and the implications they have for the future treatment of human malignancies. PMID- 16700897 TI - Introduction: the scientific and clinical rationale for extended acid suppression. PMID- 16700898 TI - Review article: the clinical pharmacology of proton pump inhibitors. AB - Proton pump inhibitors inhibit the gastric H+/K+-ATPase via covalent binding to cysteine residues of the proton pump. All proton pump inhibitors must undergo acid accumulation in the parietal cell through protonation, followed by activation mediated by a second protonation at the active secretory canaliculus of the parietal cell. The relative ease with which these steps occur with different proton pump inhibitors underlies differences in their rates of activation, which in turn influence the location of covalent binding and the stability of inhibition. Slow activation is associated with binding to a cysteine residue involved in proton transport that is located deep in the membrane. However, this is inaccessible to the endogenous reducing agents responsible for restoring H+/K+-ATPase activity, favouring a longer duration of gastric acid inhibition. Pantoprazole and tenatoprazole, a novel proton pump inhibitor which has an imidazopyridine ring in place of the benzimidazole moiety found in other proton pump inhibitors, are activated more slowly than other proton pump inhibitors but their inhibition is resistant to reversal. In addition, tenatoprazole has a greatly extended plasma half-life in comparison with all other proton pump inhibitors. The chemical and pharmacological characteristics of tenatoprazole give it theoretical advantages over benzimidazole-based proton pump inhibitors that should translate into improved acid control, particularly during the night. PMID- 16700899 TI - Review article: acid-related disease--what are the unmet clinical needs? AB - Proton pump inhibitors have dramatically improved the management options available for patients with acid-related disorders. In patients with gastro oesophageal reflux disease, currently available proton pump inhibitors provide an excellent outcome for the majority; however, they do not provide optimal pH control in many. Proton pump inhibitors co-therapy reduces, but does not eliminate, the risk of gastrointestinal ulcers and complications in patients taking non-steroidal anti-inflammatory drugs, while in patients with upper gastrointestinal bleeding, it may be difficult to reach and maintain the current therapeutic target of intragastric pH of 6-7. This article reviews the effectiveness of current antisecretory therapy in these three acid-related diseases and areas of unmet clinical need. The potential role of a proton pump inhibitor with an extended duration of action and enhanced acid control from a single daily dose, particularly improved control at night, is discussed. Finally, therapy that could be administered without regard to time of day and/or food intake would offer dosing flexibility and thus have a positive effect on patients' compliance. PMID- 16700900 TI - Review article: the opportunities and benefits of extended acid suppression. AB - Acid suppression therapy with proton pump inhibitors is associated with well established benefits in the management of gastro-oesophageal reflux (GERD) and other acid-related disorders. However, a number of issues still remain unsettled. Despite their clinical efficacy, when given once daily, currently available proton pump inhibitors may not adequately control intragastric acidity during the night in a significant proportion of both healthy subjects and GERD patients, in whom symptom relief remains suboptimal. Although some novel proton pump inhibitors have been synthesized, only few reached clinical testing. Amongst them, tenatoprazole represents a true advance displaying a long half-life (five to seven times longer than that of currently available drugs) and extended acid suppression covering both day and night. All the available clinical studies suggest both pharmacokinetic and pharmacodynamic advantages of tenatoprazole over esomeprazole. As this last compound provides - amongst the members of the class - the most effective control of intragastric pH whatever the parameter considered, it is conceivable that tenatoprazole could similarly be better than the other existing proton pump inhibitors. Tenatoprazole appears to be a promising proton pump inhibitor for the treatment of acid-related diseases, where it has the potential to address unmet clinical needs. PMID- 16700901 TI - Physiogenomic analysis of weight loss induced by dietary carbohydrate restriction. AB - BACKGROUND: Diets that restrict carbohydrate (CHO) have proven to be a successful dietary treatment of obesity for many people, but the degree of weight loss varies across individuals. The extent to which genetic factors associate with the magnitude of weight loss induced by CHO restriction is unknown. We examined associations among polymorphisms in candidate genes and weight loss in order to understand the physiological factors influencing body weight responses to CHO restriction. METHODS: We screened for genetic associations with weight loss in 86 healthy adults who were instructed to restrict CHO to a level that induced a small level of ketosis (CHO approximately 10% of total energy). A total of 27 single nucleotide polymorphisms (SNPs) were selected from 15 candidate genes involved in fat digestion/metabolism, intracellular glucose metabolism, lipoprotein remodeling, and appetite regulation. Multiple linear regression was used to rank the SNPs according to probability of association, and the most significant associations were analyzed in greater detail. RESULTS: Mean weight loss was 6.4 kg. SNPs in the gastric lipase (LIPF), hepatic glycogen synthase (GYS2), cholesteryl ester transfer protein (CETP) and galanin (GAL) genes were significantly associated with weight loss. CONCLUSION: A strong association between weight loss induced by dietary CHO restriction and variability in genes regulating fat digestion, hepatic glucose metabolism, intravascular lipoprotein remodeling, and appetite were detected. These discoveries could provide clues to important physiologic adaptations underlying the body mass response to CHO restriction. PMID- 16700902 TI - Trapping of the malaria vector Anopheles gambiae with odour-baited MM-X traps in semi-field conditions in western Kenya. AB - BACKGROUND: The successful development of odour-baited trapping systems for mosquitoes depends on the identification of behaviourally active semiochemicals, besides the design and operating principles of such devices. A large variety of 'attractants' has been identified in laboratory investigations, yet few of these increase trap catches in the field. A contained system, intermediate between the laboratory and open field, is presented and previous reports that human foot odour induces behavioural responses of Anopheles gambiae confirmed. METHODS: The response of 3-5 day old female An. gambiae towards odour-baited counterflow geometry traps (MM-X model; American Biophysics Corp., RI) was studied in semi field (screen house) conditions in western Kenya. Traps were baited with human foot odour (collected on socks), carbon dioxide (CO2, 500 ml min(-1)), ammonia (NH3), 1-octen-3-ol, or various combinations thereof. Trap catches were log (x+1) transformed and subjected to Latin square analysis of variance procedures. RESULTS: Apart from 1-octen-3-ol, all odour baits caused significant (P < 0.05) increases in trap catches over non-baited traps. Foot odour remained behaviourally active for at least 8 days after collection on nylon or cotton sock fabric. A synergistic response (P < 0.001) was observed towards the combination of foot odour and CO2, which increased catches of these odours alone by 3.8 and 2.7 times, respectively. CONCLUSION: These results are the first to report behavioural responses of an African malaria vector to human foot odour outside the laboratory, and further investigation of fractions and/or individual chemical components of this odour complex are called for. Semi-field systems offer the prospect of high-throughput screening of candidate kairomones, which may expedite the development of efficient trap-bait systems for this and other African mosquito species. PMID- 16700903 TI - Adherence of community caretakers of children to pre-packaged antimalarial medicines (HOMAPAK) among internally displaced people in Gulu district, Uganda. AB - BACKGROUND: In 2002, home-based management of fever (HBMF) was introduced in Uganda, to improve access to prompt, effective antimalarial treatment of all fevers in children under 5 years. Implementation is through community drug distributors (CDDs) who distribute pre-packaged chloroquine plus sulfadoxine pyrimethamine (HOMAPAK) free of charge to caretakers of febrile children. Adherence of caretakers to this regimen has not been studied. METHODS: A questionnaire-based survey combined with inspection of blister packaging was conducted to investigate caretakers' adherence to HOMAPAK. The population surveyed consisted of internally displaced people (IDPs) from eight camps. RESULTS: A total of 241 caretakers were interviewed. 95.0% (CI: 93.3% - 98.4%) of their children had received the correct dose for their age and 52.3% of caretakers had retained the blister pack. Assuming correct self-reporting, the overall adherence was 96.3% (CI: 93.9% - 98.7%). The nine caretakers who had not adhered had done so because the child had improved, had vomited, did not like the taste of the tablets, or because they forgot to administer the treatment. For 85.5% of cases treatment had been sought within 24 hours. Blister packaging was considered useful by virtually all respondents, mainly because it kept the drugs clean and dry. Information provided on, and inside, the package was of limited use, because most respondents were illiterate. However, CDDs had often told caretakers how to administer the treatment. For 39.4% of respondents consultation with the CDD was their reported first action when their child has fever and 52.7% stated that they consult her/him if the child does not get better. CONCLUSION: In IDP camps, the HBMF strategy forms an important component of medical care for young children. In case of febrile illness, most caretakers obtain prompt and adequate antimalarial treatment, and adhere to it. A large proportion of malaria episodes are thus likely to be treated before complications can arise. Implementation in the IDP camps now needs to focus on improving monitoring, supervision and general support to CDDs, as well as on targeting them and caretakers with educational messages. The national treatment policy for uncomplicated malaria has recently been changed to artemether-lumefantrine. Discussions on a suitable replacement combination for HBMF are well advanced, and have raised new questions about adherence. PMID- 16700905 TI - Spatio-temporal analysis of the role of climate in inter-annual variation of malaria incidence in Zimbabwe. AB - BACKGROUND: On the fringes of endemic zones climate is a major determinant of inter-annual variation in malaria incidence. Quantitative description of the space-time effect of this association has practical implications for the development of operational malaria early warning system (MEWS) and malaria control. We used Bayesian negative binomial models for spatio-temporal analysis of the relationship between annual malaria incidence and selected climatic covariates at a district level in Zimbabwe from 1988-1999. RESULTS: Considerable inter-annual variations were observed in the timing and intensity of malaria incidence. Annual mean values of average temperature, rainfall and vapour pressure were strong positive predictors of increased annual incidence whereas maximum and minimum temperature had the opposite effects. Our modelling approach adjusted for unmeasured space-time varying risk factors and showed that while year to year variation in malaria incidence is driven mainly by climate, the resultant spatial risk pattern may to large extent be influenced by other risk factors except during high and low risk years following the occurrence of extremely wet and dry conditions, respectively. CONCLUSION: Our model revealed a spatially varying risk pattern that is not attributable only to climate. We postulate that only years characterized by extreme climatic conditions may be important for developing climate based MEWS and for delineating areas prone to climate driven epidemics. However, the predictive value of climatic risk factors identified in this study still needs to be evaluated. PMID- 16700904 TI - Spatial analysis of elderly access to primary care services. AB - BACKGROUND: Admissions for Ambulatory Care Sensitive Conditions (ACSCs) are considered preventable admissions, because they are unlikely to occur when good preventive health care is received. Thus, high rates of admissions for ACSCs among the elderly (persons aged 65 or above who qualify for Medicare health insurance) are signals of poor preventive care utilization. The relevant geographic market to use in studying these admission rates is the primary care physician market. Our conceptual model assumes that local market conditions serving as interventions along the pathways to preventive care services utilization can impact ACSC admission rates. RESULTS: We examine the relationships between market-level supply and demand factors on market-level rates of ACSC admissions among the elderly residing in the U.S. in the late 1990s. Using 6,475 natural markets in the mainland U.S. defined by The Health Resources and Services Administration's Primary Care Service Area Project, spatial regression is used to estimate the model, controlling for disease severity using detailed information from Medicare claims files. Our evidence suggests that elderly living in impoverished rural areas or in sprawling suburban places are about equally more likely to be admitted for ACSCs. Greater availability of physicians does not seem to matter, but greater prevalence of non physician clinicians and international medical graduates, relative to U.S. medical graduates, does seem to reduce ACSC admissions, especially in poor rural areas. CONCLUSION: The relative importance of non-physician clinicians and international medical graduates in providing primary care to the elderly in geographic areas of greatest need can inform the ongoing debate regarding whether there is an impending shortage of physicians in the United States. These findings support other authors who claim that the existing supply of physicians is perhaps adequate, however the distribution of them across the landscape may not be optimal. The finding that elderly who reside in sprawling urban areas have access impediments about equal to residents of poor rural communities is new, and demonstrates the value of conceptualizing and modelling impedance based on place and local context. PMID- 16700906 TI - Radiation-induced sterility for pupal and adult stages of the malaria mosquito Anopheles arabiensis. AB - BACKGROUND: In the context of the Sterile Insect Technique (SIT), radiation induced sterility in the malaria mosquito Anopheles arabiensis Patton (Diptera: Culicidae) was studied. Male mosquitoes were exposed to gamma rays in the pupal or adult stage and dose-sterility curves were determined. METHODS: Pupae were irradiated shortly before emergence (at 22-26 hrs of age), and adults <24 hrs post emergence. Doses tested ranged between 0 and 100 Gy. The effects of irradiation on adult emergence, male survival, induced sterility and insemination capability were evaluated. Emergence and insemination data were analysed using independent t-tests against the control. Correlation analyses were performed for insemination rate and dose and insemination and fecundity. Male survival was analysed using Kaplan-Meier survival analyses. Finally, the calculated residual fertility values were inverse-normal transformed and linear regression analyses performed. RESULTS: Irradiation of pupae, for all doses tested, had no effect on adult emergence. Survival curves of males irradiated as pupae or adults were similar or even slightly higher than non-irradiated males. Overall, adults appeared to be slightly more susceptible to irradiation, although no significant differences for individual doses were observed. In the pupal stage, a significant negative correlation was found between insemination and dose, but the correlation coefficient was associated with less than 25% of the total variation. A review of the literature indicated that An. arabiensis is more radiation resistant than other anopheline mosquitoes. CONCLUSION: The optimal dose for male insects to be released in an SIT programme depends on their level of sterility and competitiveness. The use of semi-sterilizing doses to produce more competitive insects is discussed. The most convenient developmental stage for mosquito irradiation on a mass-scale are pupae, but pupal irradiation resulted in a lower insemination rate at the highest dose compared to adult irradiation. On the basis of this study, a suitable dose range that includes semi-sterilizing doses is identified to initiate competitiveness experiments for males irradiated at both developmental stages. PMID- 16700907 TI - Environmental influences on energy balance-related behaviors: a dual-process view. AB - BACKGROUND: Studies on the impact of the 'obesogenic' environment have often used non-theoretical approaches. In this journal's debate and in other papers authors have argued the necessity of formulating conceptual models for differentiating the causal role of environmental influences on behavior. DISCUSSION: The present paper aims to contribute to the debate by presenting a dual-process view on the environment--behavior relationship. This view is conceptualized in the EnRG framework (Environmental Research framework for weight Gain prevention). In the framework, behavior is postulated to be the result of a simultaneous influence of conscious and unconscious processes. Environmental influences are hypothesized to influence behavior both indirectly and directly. The indirect causal mechanism reflects the mediating role of behavior-specific cognitions in the influence of the environment on behavior. A direct influence reflects the automatic, unconscious, influence of the environment on behavior. Specific personal and behavioral factors are postulated to moderate the causal path (i.e., inducing either the automatic or the cognitively mediated environment - behavior relation). In addition, the EnRG framework applies an energy balance-approach, stimulating the integrated study of determinants of diet and physical activity. CONCLUSION: The application of a dual-process view may guide research towards causal mechanisms linking specific environmental features with energy balance related behaviors in distinct populations. The present paper is hoped to contribute to the evolution of a paradigm that may help to disentangle the role of 'obesogenic' environmental factors. PMID- 16700908 TI - Gene-enhanced tissue engineering for dental hard tissue regeneration: (1) overview and practical considerations. AB - Gene-based therapies for tissue regeneration involve delivering a specific gene to a target tissue with the goal of changing the phenotype or protein expression profile of the recipient cell; the ultimate goal being to form specific tissues required for regeneration. One of the principal advantages of this approach is that it provides for a sustained delivery of physiologic levels of the growth factor of interest.This manuscript will review the principals of gene-enhanced tissue engineering and the techniques of introducing DNA into cells. Part 2 will review recent advances in gene-based therapies for dental hard tissue regeneration, specifically as it pertains to dentin regeneration/pulp capping and periodontal regeneration. PMID- 16700910 TI - International Rotifer Symposia: prospects and retrospects from Rotifera XI. AB - The XI International Rotifer Symposium was held during 11-18 March, 2006 at the National Autonomous University of Mexico Campus Iztacala located at the North Mexico City (Mexico). These triennial international meetings, first organized in Austria by Late Ruttner-Kolisko in September 1976, are gradually becoming the focal point of discussion and collaboration from rotifer workers across the world. The present XI symposium was attended by 125 participants from 20 nations. During this meeting, different themes of rotifer research from morphology to molecular biology were considered. In addition, there were four invited lectures and four workshops covering different themes of the symposium. During the last 30 years, rotifer research has witnessed gradual shift from the conventional morphological taxonomy to molecular and evolutionary systematics. While the basic rotifer ecological studies continue today, applied areas such as ecotoxicology and aquaculture have taken key roles in the recent meetings. The international rotifer meetings provide ample opportunities not only for exchange of ideas and recent research, but also for material and in establishing inter-personal relationships. Over the last 30 years, the number of participants attending the rotifer meetings has increased. PMID- 16700909 TI - Promoter hypermethylation-mediated inactivation of multiple Slit-Robo pathway genes in cervical cancer progression. AB - BACKGROUND: Cervical Cancer (CC) exhibits highly complex genomic alterations. These include hemizygous deletions at 4p15.3, 10q24, 5q35, 3p12.3, and 11q24, the chromosomal sites of Slit-Robo pathway genes. However, no candidate tumor suppressor genes at these regions have been identified so far. Slit family of secreted proteins modulates chemokine-induced cell migration of distinct somatic cell types. Slit genes mediate their effect by binding to its receptor Roundabout (Robo). These genes have shown to be inactivated by promoter hypermethylation in a number of human cancers. RESULTS: To test whether Slit-Robo pathway genes are targets of inactivation at these sites of deletion, we examined promoter hypermethylation of SLIT1, SLIT2, SLIT3, ROBO1, and ROBO3 genes in invasive CC and its precursor lesions. We identified a high frequency of promoter hypermethylation in all the Slit-Robo genes resulting in down regulated gene expression in invasive CC, but the inhibitors of DNA methylation and histone deacetylases (HDACs) in CC cell lines failed to effectively reactivate the down regulated expression. These results suggest a complex mechanism of inactivation in the Slit-Robo pathway in CC. By analysis of cervical precancerous lesions, we further show that promoter hypermethylation of Slit-Robo pathway occurs early in tumor progression. CONCLUSION: Taken together, these findings suggest that epigenetic alterations of Slit-Robo pathway genes (i) play a role in CC development, (ii) further delineation of molecular basis of promoter methylation mediated gene regulation provides a potential basis for epigenetic-based therapy in advanced stage CC, and (iii) form epigenetic signatures to identify precancerous lesions at risk to progression. PMID- 16700911 TI - The clinical relevance of the metabolism of prostate cancer; zinc and tumor suppression: connecting the dots. AB - BACKGROUND: The genetic and molecular mechanisms responsible for and associated specifically with the development and progression of malignant prostate cells are largely unidentified. In addition, despite its implication in virtually all malignant cells, the role of altered cellular metabolism as an essential factor in prostate malignancy has been largely ignored. Moreover, the intermediary metabolism of normal prostate as well as malignant prostate cells is among the least studied and most poorly understood of all mammalian cells. Some important factors, especially the role of zinc, have been identified and implicated in the development and progression of prostate malignancy. In this review, we provide a current and updated integrated assessment of the relationships of intermediary metabolism in normal prostate and in prostate cancer. The experimental and clinical evidence that leads to the formulation of concepts of normal and malignant prostate metabolism is presented. The evidence for a concept of zinc as a tumor suppressor agent and Zip1 zinc transporter as a tumor-suppressor gene is described. RESULTS: The specialized function of the normal prostate glandular epithelium to produce and secrete enormously high levels of citrate involves and requires unique intermediary metabolism activities that are not generally associated with other normal mammalian cells. The accumulation of zinc by these cells is an essential factor in this unique metabolic relationship. In malignancy, the normal zinc-accumulating citrate-producing epithelial cells are metabolically transformed to citrate-oxidizing cells that lose the ability to accumulate zinc. A genetic alteration in the expression of ZIP1 zinc transporter is associated with this metabolic transformation. These genetic/metabolic relationships have important consequences on citrate-related metabolism, bioenergetics, cell proliferation and invasive capabilities of the malignant cells, which result in tumor-suppression characteristics. CONCLUSION: The genetic/metabolic relationships in normal prostate glandular epithelium are driven by the unique function to accumulate and secrete citrate. The genetic/metabolic transformation of the prostate malignant cells is driven by the metabolic/bioenergetic, growth/proliferative, and invasive/migration requirements of the malignant process. Zinc is critical to these relationships. An understanding of these genetic/metabolic relationships provides new directions and opportunities for development of regimens for the prevention and treatment of prostate cancer. Important insight into the genetic/metabolic requirements of the prostate malignant process is now evolving. Most importantly at this time, an appreciation and recognition of the genetic/metabolic significance and implications in the development of prostate malignancy is imperative; and much needed research in this area is essential. Hopefully, this review will help to achieve these goals. PMID- 16700912 TI - Double blind, randomized controlled trial, to evaluate the effectiveness of a controlled nitric oxide releasing patch versus meglumine antimoniate in the treatment of cutaneous leishmaniasis [NCT00317629]. AB - BACKGROUND: Cutaneous Leishmaniasis is a worldwide disease, endemic in 88 countries, that has shown an increasing incidence over the last two decades. So far, pentavalent antimony compounds have been considered the treatment of choice, with a percentage of cure of about 85%. However, the high efficacy of these drugs is counteracted by their many disadvantages and adverse events. Previous studies have shown nitric oxide to be a potential alternative treatment when administered topically with no serious adverse events. However, due to the unstable nitric oxide release, the topical donors needed to be applied frequently, making the adherence to the treatment difficult. The electrospinning technique has allowed the production of a multilayer transdermal patch that produces a continuous and stable nitric oxide release. The main objective of this study is to evaluate this novel nitric oxide topical donor for the treatment of cutaneous leishmaniasis. METHODS AND DESIGN: A double-blind, randomized, double-masked, placebo-controlled clinical trial, including 620 patients from endemic areas for Leishmaniasis in Colombia was designed to investigate whether this patch is as effective as meglumine antimoniate for the treatment of cutaneous leishmaniasis but with less adverse events. Subjects with ulcers characteristic of cutaneous leishmaniasis will be medically evaluated and laboratory tests and parasitological confirmation performed. After checking the inclusion/exclusion criteria, the patients will be randomly assigned to one of two groups. During 20 days Group 1 will receive simultaneously meglumine antimoniate and placebo of nitric oxide patches while Group 2 will receive placebo of meglumine antimoniate and active nitric oxide patches. During the treatment visits, the medications will be daily administered and the presence of adverse events assessed. During the follow-up, the research group will visit the patients at days 21, 45, 90 and 180. The healing process of the ulcer, the health of the participants, recidivisms and/or reinfection will also be assessed. The evolution of the ulcers will be photographically registered. In case that the effectiveness of the patches is demonstrated, a novel and safe therapeutic alternative for one of the most important public health problems in many countries will be available to patients. PMID- 16700913 TI - Malaria in Sri Lanka: one year post-tsunami. AB - One year ago, the authors of this article reported in this journal on the malaria situation in Sri Lanka prior to the tsunami that hit on 26 December 2004, and estimated the likelihood of a post-tsunami malaria outbreak to be low. Malaria incidence has decreased in 2005 as compared to 2004 in most districts, including the ones that were hit hardest by the tsunami. The malaria incidence (aggregated for the whole country) in 2005 followed the downward trend that started in 2000. However, surveillance was somewhat affected by the tsunami in some coastal areas and the actual incidence in these areas may have been higher than recorded, although there were no indications of this and it is unlikely to have affected the overall trend significantly. The focus of national and international post tsunami malaria control efforts was supply of antimalarials, distribution of impregnated mosquito nets and increased monitoring in the affected area. Internationally donated antimalarials were either redundant or did not comply with national drug policy, however, few seem to have entered circulation outside government control. Despite distribution of mosquito nets, still a large population is relatively exposed to mosquito bites due to inadequate housing. There were no indications of increased malaria vector abundance. Overall it is concluded that the tsunami has not negatively influenced the malaria situation in Sri Lanka. PMID- 16700914 TI - Resveratrol, but not EGCG, in the diet suppresses DMBA-induced mammary cancer in rats. AB - Despite the advent of new and aggressive therapeutics, breast cancer remains a leading killer among women; hence there is a need for the prevention of this disease. Several naturally occurring polyphenols have received much attention for their health benefits, including anti-carcinogenic properties. Two of these are resveratrol, a component of red grapes, and epigallocatechin-3-gallate (EGCG), the major catechin found in green tea. In this study, we tested the hypothesis that these two polyphenols protect against chemically-induced mammary cancer by modulating mammary gland architecture, cell proliferation, and apoptosis. Female Sprague-Dawley CD rats were exposed to either resveratrol (1 g/kg AIN-76A diet), EGCG (0.065% in the drinking water), or control diet (AIN-76A) for the entirety of their life starting at birth. At 50 days postpartum, rats were treated with 60 mg dimethylbenz[a]anthracene (DMBA)/kg body weight to induce mammary cancer. Resveratrol, but not EGCG, suppressed mammary carcinogenesis (fewer tumors per rat and longer tumor latency). Analysis of mammary whole mounts from 50-day-old rats revealed that resveratrol, but not EGCG, treatment resulted in more differentiated lobular structures. Bromodeoxyuridine (BrdU) incorporation studies showed that resveratrol treatment caused a significant reduction in proliferative cells in mammary terminal ductal structures at 50 days postpartum, making them less susceptible to carcinogen insult. The epithelial cells of terminal end buds in the mammary glands of resveratrol-treated rats also showed an increase in apoptotic cells compared to the control or EGCG-treated rats as measured by a DNA fragmentation assay. At the given doses, resveratrol treatment resulted in a serum resveratrol concentration of 2.00 microM, while treatment with EGCG resulted in a serum EGCG concentration of 31.06 nM. 17beta-Estradiol, progesterone, and prolactin concentrations in the serum were not significantly affected by resveratrol or EGCG. Neither polyphenol treatment resulted in toxicity as tested by alterations in body weights, diet and drink consumptions, and day to vaginal opening. We conclude that resveratrol in the diet can reduce susceptibility to mammary cancer, while EGCG in the drinking water at the dose used was not effective. PMID- 16700915 TI - Genetic variants of adiponectin receptor 2 are associated with increased adiponectin levels and decreased triglyceride/VLDL levels in patients with metabolic syndrome. AB - BACKGROUND: Adiponectin acts as an antidiabetic, antiinflammatory and antiatherogenic adipokine. These effects are assumed to be mediated by the recently discovered adiponectin receptors AdipoR1 and AdipoR2. AIM: The purpose of this study was to determine whether variations in the AdipoR1 and AdipoR2 genes may contribute to insulin resistance, dyslipidemia and inflammation. METHODS: We sequenced all seven coding exons of both genes in 20 unrelated German subjects with metabolic syndrome and tested genetic variants for association with glucose, lipid and inflammatory parameters. RESULTS: We identified three AdipoR2 variants (+795G/A, +870C/A and +963C/T) in perfect linkage disequilibrium (r2 = 1) with a minor allele frequency of 0.125. This haplotype was associated with higher plasma adiponectin levels and decreased fasting triglyceride, VLDL triglyceride and VLDL-cholesterol levels. No association, however, was observed between the AdipoR2 SNP cluster and glucose metabolism. CONCLUSION: To our knowledge, this is the first study to identify an association between genetic variants of the adiponectin receptor genes and plasma adiponectin levels. Furthermore, our data suggest that AdipoR2 may play an important role in triglyceride/VLDL metabolism. PMID- 16700917 TI - On the typology and the worship status of sacred trees with a special reference to the Middle East. AB - This article contains the reasons for the establishment of sacred trees in Israel based on a field study. It includes 97 interviews with Muslim and Druze informants. While Muslims (Arabs and Bedouins) consider sacred trees especially as an abode of righteous figures' (Wellis') souls or as having a connection to their graves, the Druze relate sacred trees especially to the events or deeds in the lives of prophets and religious leaders. A literary review shows the existence of 24 known reasons for the establishment of sacred trees worldwide, 11 of which are known in Israel one of these is reported here for the first time. We found different trends in monotheistic and polytheistic religions concerning their current worship of sacred trees. PMID- 16700919 TI - Motion and twisting of magnetic particles ingested by alveolar macrophages in the human lung: effect of smoking and disease. AB - BACKGROUND: Magnetic microparticles being ingested by alveolar macrophages can be used as a monitor for intracellular phagosome motions and cytoskeletal mechanical properties. These studies can be performed in the human lung after voluntary inhalation. The influence of cigarette smoking and lung diseases on cytoskeleton dependent functions was studied. METHODS: Spherical 1.3 microm diameter ferrimagnetic iron oxide particles were inhaled by 17 healthy volunteers (40-65 years), 15 patients with sarcoidosis (SAR), 12 patients with idiopathic pulmonary fibrosis (IPF), and 18 patients with chronic obstructive bronchitis (COB). The retained particles were magnetized and aligned in an external 100 mT magnetic field. All magnetized particles induce a weak magnetic field of the lung, which was detected by a sensitive SQUID (superconducting quantum interference device) sensor. Cytoskeletal reorganizations within macrophages and intracellular transport cause stochastic magnetic dipole rotations, which are reflected in a decay of the magnetic lung field, called relaxation. Directed phagosome motion was induced in a weak magnetic twisting field. The resistance of the cytoplasm to particle twisting was characterized by the viscosity and the stiffness (ratio between stress to strain) of the cytoskeleton. RESULTS: One week after particle inhalation and later macrophage motility (relaxation) and cytoskeletal stiffness was not influenced by cigarette smoking, neither in healthy subjects, nor in the patients. Patients with IPF showed in tendency a faster relaxation (p = 0.06). Particle twisting revealed a non-Newtonian viscosity with a pure viscous and a viscoelastic compartment. The viscous shear was dominant, and only 27% of the shear recoiled and reflected viscoelastic properties. In patients with IPF, the stiffness was reduced by 60% (p < 0.02). An analysis of the shear rate and stress dependence of particle twisting allows correlating the rheological compartments to cytoskeletal subunits, in which microtubules mediate the pure viscous (non recoverable) shear and microfilaments mediate the viscoelastic (recoverable) behavior. The missing correlation between relaxation and particle twisting shows that both stochastic and directed phagosome motion reflect different cytoskeletal mechanisms. CONCLUSION: Faster relaxation and a soft cytoskeleton in patients with IPF indicate alterations in cytoskeleton dependent functions of alveolar macrophages, which may cause dysfunction's in the alveolar defense, like a slower migration, a retarded phagocytosis, a disturbed phagosome lysosome fusion and an impaired clearance. PMID- 16700916 TI - Caerulein-induced acute pancreatitis in mice that constitutively overexpress Reg/PAP genes. AB - BACKGROUND: The cystic fibrosis (CF) mouse pancreas has constitutively elevated expression of the Reg/PAP cell stress genes (60-fold greater Reg3alpha, and 10 fold greater PAP/Reg3beta and Reg3gamma). These genes are suggested to be involved in protection or recovery from pancreatic injury. METHODS: To test this idea the supramaximal caerulein model was used to induce acute pancreatitis in wild type and CF mice. Serum amylase, pancreatic water content (as a measure of edema), pancreatic myeloperoxidase activity, and Reg/PAP expression were quantified. RESULTS: In both wild type and CF mice caerulein induced similar elevations in serum amylase (maximal at 12 h), pancreatic edema (maximal at 7 h), and pancreatic myeloperoxidase activity (MPO, a marker of neutrophil infiltration; maximal at 7 h). By immunohistochemistry, Reg3alpha was strongly expressed in the untreated CF pancreas but not in wild type. During pancreatitis, Reg3alpha was intensely expressed in foci of inflamed tissue in both wild type and CF. CONCLUSION: These data demonstrate that the severity of caerulein-induced pancreatitis is not ameliorated in the CF mouse even though the Reg/PAP stress genes are already highly upregulated. While Reg/PAP may be protective they may also have a negative effect during pancreatitis due to their anti-apoptotic activity, which has been shown to increase the severity of pancreatitis. PMID- 16700918 TI - Response of spontaneously hypertensive rats to inhalation of fine and ultrafine particles from traffic: experimental controlled study. AB - BACKGROUND: Many epidemiological studies have shown that mass concentrations of ambient particulate matter (PM) are associated with adverse health effects in the human population. Since PM is still a very crude measure, this experimental study has explored the role of two distinct size fractions: ultrafine (<0.15 microm) and fine (0.15- 2.5 microm) PM. In a series of 2-day inhalation studies, spontaneously hypersensitive (SH) rats were exposed to fine, concentrated, ambient PM (fCAP) at a city background location or a combination of ultrafine and fine (u+fCAP) PM at a location dominated by traffic. We examined the effect on inflammation and both pathological and haematological indicators as markers of pulmonary and cardiovascular injury. Exposure concentrations ranged from 399 microg/m3 to 3613 microg/m3 for fCAP and from 269 microg/m3 to 556 microg/m3 for u+fCAP. RESULTS: Ammonium, nitrate, and sulphate ions accounted for 56 +/- 16% of the total fCAP mass concentrations, but only 17 +/- 6% of the u+fCAP mass concentrations. Unambiguous particle uptake in alveolar macrophages was only seen after u+fCAP exposures. Neither fCAP nor u+fCAP induced significant changes of cytotoxicity or inflammation in the lung. However, markers of oxidative stress (heme oxygenase-1 and malondialdehyde) were affected by both fCAP and u+fCAP exposure, although not always significantly. Additional analysis revealed heme oxygenase-1 (HO-1) levels that followed a nonmonotonic function with an optimum at around 600 microg/m3 for fCAP. As a systemic response, exposure to u+fCAP and fCAP resulted in significant decreases of the white blood cell concentrations. CONCLUSION: Minor pulmonary and systemic effects are observed after both fine and ultrafine + fine PM exposure. These effects do not linearly correlate with the CAP mass. A greater component of traffic CAP and/or a larger proportion ultrafine PM does not strengthen the absolute effects. PMID- 16700920 TI - Bounded search for de novo identification of degenerate cis-regulatory elements. AB - BACKGROUND: The identification of statistically overrepresented sequences in the upstream regions of coregulated genes should theoretically permit the identification of potential cis-regulatory elements. However, in practice many cis-regulatory elements are highly degenerate, precluding the use of an exhaustive word-counting strategy for their identification. While numerous methods exist for inferring base distributions using a position weight matrix, recent studies suggest that the independence assumptions inherent in the model, as well as the inability to reach a global optimum, limit this approach. RESULTS: In this paper, we report PRISM, a degenerate motif finder that leverages the relationship between the statistical significance of a set of binding sites and that of the individual binding sites. PRISM first identifies overrepresented, non degenerate consensus motifs, then iteratively relaxes each one into a high scoring degenerate motif. This approach requires no tunable parameters, thereby lending itself to unbiased performance comparisons. We therefore compare PRISM's performance against nine popular motif finders on 28 well-characterized S. cerevisiae regulons. PRISM consistently outperforms all other programs. Finally, we use PRISM to predict the binding sites of uncharacterized regulons. Our results support a proposed mechanism of action for the yeast cell-cycle transcription factor Stb1, whose binding site has not been determined experimentally. CONCLUSION: The relationship between statistical measures of the binding sites and the set as a whole leads to a simple means of identifying the diverse range of cis-regulatory elements to which a protein binds. This approach leverages the advantages of word-counting, in that position dependencies are implicitly accounted for and local optima are more easily avoided. While we sacrifice guaranteed optimality to prevent the exponential blowup of exhaustive search, we prove that the error is bounded and experimentally show that the performance is superior to other methods. A Java implementation of this algorithm can be downloaded from our web server at http://genie.dartmouth.edu/prism. PMID- 16700922 TI - Cortical activation elicited by unrecognized stimuli. AB - BACKGROUND: It is unclear whether a stimulus that cannot be recognized consciously, could elicit a well-processed cognitive response. METHODS: We used functional imaging to examine the pattern of cortical activation elicited by unrecognized stimuli during memory processing. Subjects were given a recognition task using recognizable and non-recognizable subliminal stimuli. RESULTS: Unrecognized stimuli activated the cortical areas that are associated with retrieval attempt (left prefrontal), and novelty detection (left hippocampus). This indicates that the stimuli that were not consciously recognized, activated neural network associated with aspects of explicit memory processing. CONCLUSION: Results suggest that conscious recognition of stimuli is not necessary for activation of cognitive processing. PMID- 16700921 TI - Contribution of alpha- and beta-defensins to lung function decline and infection in smokers: an association study. AB - BACKGROUND: Alpha-defensins, which are major constituents of neutrophil azurophilic granules, and beta-defensins, which are expressed in airway epithelial cells, could contribute to the pathogenesis of chronic obstructive pulmonary disease by amplifying cigarette smoke-induced and infection-induced inflammatory reactions leading to lung injury. In Japanese and Chinese populations, two different beta-defensin-1 polymorphisms have been associated with chronic obstructive pulmonary disease phenotypes. We conducted population based association studies to test whether alpha-defensin and beta-defensin polymorphisms influenced smokers' susceptibility to lung function decline and susceptibility to lower respiratory infection in two groups of white participants in the Lung Health Study (275 = fast decline in lung function and 304 = no decline in lung function). METHODS: Subjects were genotyped for the alpha defensin-1/alpha-defensin-3 copy number polymorphism and four beta-defensin-1 polymorphisms (G-20A, C-44G, G-52A and Val38Ile). RESULTS: There were no associations between individual polymorphisms or imputed haplotypes and rate of decline in lung function or susceptibility to infection. CONCLUSION: These findings suggest that, in a white population, the defensin polymorphisms tested may not be of importance in determining who develops abnormally rapid lung function decline or is susceptible to developing lower respiratory infections. PMID- 16700923 TI - Gene expression alterations in brains of mice infected with three strains of scrapie. AB - BACKGROUND: Transmissible spongiform encephalopathies (TSEs) or prion diseases are fatal neurodegenerative disorders which occur in humans and various animal species. Examples include Creutzfeldt-Jakob disease (CJD) in humans, bovine spongiform encephalopathy (BSE) in cattle, chronic wasting disease (CWD) in deer and elk, and scrapie in sheep, and experimental mice. To gain insights into TSE pathogenesis, we made and used cDNA microarrays to identify disease-associated alterations in gene expression. Brain gene expression in scrapie-infected mice was compared to mock-infected mice at pre-symptomatic and symptomatic time points. Three strains of mouse scrapie that show striking differences in neuropathology were studied: ME7, 22L, and Chandler/RML. RESULTS: In symptomatic mice, over 400 significant gene expression alterations were identified. In contrast, only 22 genes showed significant alteration in the pre-symptomatic animals. We also identified genes that showed significant differences in alterations in gene expression between strains. Genes identified in this study encode proteins that are involved in many cellular processes including protein folding, endosome/lysosome function, immunity, synapse function, metal ion binding, calcium regulation and cytoskeletal function. CONCLUSION: These studies shed light on the complex molecular events that occur during prion disease, and identify genes whose further study may yield new insights into strain specific neuropathogenesis and ante-mortem tests for TSEs. PMID- 16700924 TI - Comparison of tricuspid inflow and superior vena caval Doppler velocities in acute simulated hypovolemia: new non-invasive indices for evaluating right ventricular preload. AB - BACKGROUND: Assessment of cardiac preload is important for clinical management of some emergencies related to hypovolemia. Effects of acute simulated hypovolemia on Doppler blood flow velocity indices of tricuspid valve (TV) and superior vena cava (SVC) were investigated in order to find sensitive Doppler indices for predicting right ventricular preload. METHODS: Doppler flow patterns of SVC and TV in 12 healthy young men were examined by transthoracic echocardiography (TTE) during graded lower body negative pressure (LBNP) of up to -60 mm Hg which simulated acute hypovolemia. Peak velocities of all waves and their related ratios (SVC S/D and tricuspid E/A) were measured, calculated and statistically analyzed. RESULTS: Except for the velocity of tricuspid A wave, velocities of all waves and their related ratios declined during volume decentralization. Of all indices measured, the peak velocities of S wave and AR wave in SVC correlated most strongly with levels of LBNP (r = -0.744 and -0.771, p < 0.001). CONCLUSION: The S and AR velocities are of good values in assessing right ventricular preload. Monitoring SVC flow may provide a relatively noninvasive means to assess direct changes in right ventricular preload. PMID- 16700925 TI - Generation of H9 T-cells stably expressing a membrane-bound form of the cytoplasmic tail of the Env-glycoprotein: lack of transcomplementation of defective HIV-1 virions encoding C-terminally truncated Env. AB - H9-T-cells do not support the replication of mutant HIV-1 encoding Env protein lacking its long cytoplasmic C-terminal domain (Env-CT). Here we describe the generation of a H9-T-cell population constitutively expressing the HIV-1 Env-CT protein domain anchored in the cellular membrane by it homologous membrane spanning domain (TMD). We confirmed that the Env-TMD-CT protein was associated with cellular membranes, that its expression did not have any obvious cytotoxic effects on the cells and that it did not affect wild-type HIV-1 replication. However, as measured in both a single-round assay as well as in spreading infections, replication competence of mutant pNL-Tr712, lacking the Env-CT, was not restored in this H9 T-cell population. This means that the Env-CT per se cannot transcomplement the replication block of HIV-1 virions encoding C terminally truncated Env proteins and suggests that the Env-CT likely exerts its function only in the context of the complete Env protein. PMID- 16700926 TI - Tuberous sclerosis: genes, brain, and behaviour. PMID- 16700928 TI - Trends in the prevalence of cerebral palsy in Northern Ireland, 1981-1997. AB - We describe trends in the prevalence of cerebral palsy (CP) by birthweight group, and in the severity of motor impairments and presence of associated intellectual impairment, in Northern Ireland from 1981 to 1997 (n=909; 510 males, 399 females; total population 415,936 live births) using data from a population-based register of CP. Children with suspected CP or who died before 1 year of age and those with CP of postneonatal origin were excluded. Prevalence of CP was 2.2 per 1,000 live births without significant change over time. Among very-low-birthweight (<1,500 g) live births, prevalence was 44.5 per 1,000 (95% confidence interval 32.3-59.8) from 1994 to 1997, with evidence of a statistically significant decline in prevalence since the mid- to late 1980s accompanied by a decrease in the severity of motor impairment and likelihood of intellectual impairment. Among moderately low-birthweight (1,500-2,499 g) children there was weaker evidence of a peak prevalence in the late 1980s. Prevalence among normal-birthweight infants did not change significantly, but outcome in terms of severity of motor impairment and intellectual impairment improved in the 1990s. Occurrence of bilateral spasticity from 1994 to 1997 was associated with greater severity and likelihood of intellectual impairment for normal-birthweight individuals than for low- or very low-birthweight individuals. PMID- 16700929 TI - Prevalence of cerebral palsy in Turkish children between the ages of 2 and 16 years. AB - The aim of this cross-sectional study was to determine the prevalence of cerebral palsy (CP) among children in Turkey between the ages of 2 and 16 years. Samples were selected from cities, towns, districts, and villages using the cluster sampling method; 41,861 children were selected. Data was collected by parental interview and physical examination. One hundred and eighty-six children were identified with CP. The prevalence of CP was determined as 4.4 per 1,000 live births and included postnatally acquired CP. Origin of CP was classified as prenatal in 49 (26.6%), perinatal/neonatal in 34 (18.5%), postnatal in 11 (5.9%), and unclassifiable in 90 participants (48.9%; data was unobtainable for two individuals). Type of CP was diplegia in 39.8% of children, hemiplegia in 28%, tetraplegia in 19.9%, ataxia in 5.9%, and dyskinetic in 6.4%. Prenatal factors were seen more frequently in the groups with a high socioeconomic status while perinatal factors were encountered more often in those with a low socioeconomic status (p<0.05). Place of residence and sex had no significant effect on the prevalence of CP (p>0.05). This cross-sectional study shows that the prevalence of CP in Turkey is higher than that in developed countries but the aetiology is probably similar. Although the high prevalence of CP in Turkey could originate from an increased level of obstetric and neonatal problems, the lack of a possible aetiological factor in approximately half the children suggests that the high level might also be due to other factors, such as genetic disorders. PMID- 16700930 TI - Gross and fine motor function and accompanying impairments in cerebral palsy. AB - The aim of this study was to describe and analyze gross and fine motor function and accompanying neurological impairments in children with cerebral palsy (CP) born between 1991 and 1998 in western Sweden. A population-based study comprised 411 children with a diagnosis of CP ascertained at 4 to 8 years of age. Gross Motor Function Classification System (GMFCS) levels were documented in 367 children (205 males, 162 females). Bimanual Fine Motor Function (BFMF) classification levels of 345 of the children and information on learning disability, epilepsy, visual and hearing impairments, and hydrocephalus from 353 children were obtained. For spastic CP, a new classification according to the Surveillance of Cerebral Palsy in Europe of uni- and bilateral spastic CP was applied. GMFCS was distributed at Level I in 32%, Level II in 29%, Level III in 8%, Level IV in 15%, and Level V in 16%. The corresponding percentages for BFMF were 30.7%, 31.6%, 12.2%, 11.9%, and 13.6% respectively. Learning disability was present in 40%, epilepsy in 33%, and severe visual impairment in 19% of the children. Motor function differed between CP types. More severe GMFCS levels correlated with larger proportions of accompanying impairments and, in children born at term, to the presence of adverse peri/neonatal events in the form of intracranial haemorrhage/stroke, cerebral infection, and hypoxic-ischaemic encephalopathy. GMFCS Level I correlated positively to increasing gestational age. We conclude that the classification of CP should be based on CP type and motor function, as the two combine to produce an indicator of total impairment load. PMID- 16700932 TI - Clinician agreement on gait pattern ratings in children with spastic hemiplegia. AB - The level of agreement between clinicians' ratings of gait patterns in children with spastic hemiplegia was investigated using the Winters, Gage, and Hicks (WGH) classification scale. Sixteen clinicians (nine physiotherapists, seven orthopaedic surgeons) from six gait laboratories in five different countries rated gait patterns in 34 children with spastic hemiplegia (22 males, 12 females; mean age 10y [SD 3y], range 6-18y). Archived three-dimensional gait reports (kinematic and video data) and videos (video clips only) were rated for each child. Agreement between clinicians was substantial for reports (weighted kappa [wkappa] 0.77, range 0.62-0.89). Agreement was lower for videos (wkappa 0.63, range 0.39-0.97) and for clinicians' agreement between reports and corresponding videos (wkappa 0.62, range 0.47-0.76). Exact agreement was unacceptable for some gait patterns using reports (mean 65%, range 32-74%) and videos (mean 53%, range 35-94%). Not all gait patterns could be rated using the original WGH categories. It was concluded that: (1) agreement is acceptable using the WGH tool; (2) kinematic data from 3D instrumented gait analysis and video should be used together when using the WGH scale; and (3) further refinement of this classification is required. PMID- 16700931 TI - Stability of the gross motor function classification system. AB - The aim of this study was to assess the stability of the Gross Motor Function Classification System (GMFCS) by examining whether children with cerebral palsy (CP) remain in the same level over time. Participants were 610 children with CP (342 males, 268 females; mean age 6y 9mo [SD 2y 10mo]), range 16mo-13y). Children were assessed 2 to 7 times (mean 4.3) at 6-month (children <6y old) or 12 month(children >or=6y old) intervals. Seventy-three per cent of children remained in the same level for all ratings. The weighted kappa coefficient between the first and last ratings was 0.84 for children less than 6 years old and 0.89 for children at least 6 years old, indicating excellent chance-corrected agreement. Children initially classified in Levels I and V were least likely to be reclassified. There was a tendency for children younger than 6 years who were reclassified to be done so to a lower level of ability. The results provide evidence of stability of the GMFCS. PMID- 16700933 TI - Movement patterns of the upper extremity and trunk before and after corrective surgery of impaired forearm rotation in patients with cerebral palsy. AB - The effect of surgical correction of impaired forearm rotation on associated body movement patterns was studied prospectively by comparison of preoperative and postoperative three-dimensional video analysis of the upper extremity and trunk in eight male and two female patients with hemiplegic cerebral palsy (CP; mean age 16y 2mo [SD 4y 11mo]; range 11-27y). A customized parameter, 'extrinsic forearm rotation', was used to quantify associated movements supplementing forearm rotation. After surgical correction of the pronation deformity, active forearm supination during a functional reaching task had improved by a mean of 37 degrees in combination with significantly decreased extrinsic forearm rotation by a mean of 13 degrees . In addition, an average loss of 16 degrees of active pronation in combination with increased extrinsic forearm rotation (mean 8 degrees ) was observed. On the basis of these results we conclude that successful surgical correction of a pronation deformity in patients with CP directly affects related movement patterns of the upper extremity and trunk. PMID- 16700934 TI - The PedsQL in pediatric cerebral palsy: reliability, validity, and sensitivity of the Generic Core Scales and Cerebral Palsy Module. AB - This investigation determined the measurement properties of the Pediatric Quality of Life Inventory (PedsQL) 3.0 Cerebral Palsy (CP) Module. PedsQL 4.0 Generic Core Scales and 3.0 CP Module were administered to 245 families. Mean age of the 134 males and 107 females was 8 years 1 month (SD 4y 4mo; range 2-18y). The sample included children with hemiplegia (n=55), diplegia (n=84), and quadriplegia (n=85). Twenty eight children had a Gross Motor Function Classification System classification at Level I, 40 at Level II, 86 at Level III, 43 at Level IV, and 37 at Level V. Reliability was demonstrated for the PedsQL 4.0 (alpha=0.86 child, 0.89 parent) and CP Module (alpha=0.79 child, 0.91 parent). The PedsQL 4.0 distinguished between healthy children and children with CP. Construct validity of the CP Module was supported. Sensitivity of the PedsQL was demonstrated among children with different diagnostic categories and gross motor function. PMID- 16700935 TI - Gram-negative meningitis and infections in individuals treated with intrathecal baclofen for spasticity: a retrospective study. AB - The aim of this retrospective study was to describe signs, symptoms, and clinical outcomes of individuals undergoing intrathecal baclofen (ITB) therapy who experienced pump-related Gram-negative infections including meningitis. Participants included 12 individuals (nine males, three females) aged 10 to 32 years (mean 17y 9mo), nine of whom had quadriplegic CP. A total of 571 baclofen pump surgeries were performed with 45 total infections. Of the 45 infections, 12 were by Gram-negative organisms, two resulting in meningitis. Ten of 12 Gram negative infections (21 site encounters) occurred within 60 days of surgery. Eleven of 12 pumps were explanted. By site encounters, Pseudomonas aeruginosa accounted for eight Gram-negative infections, Escherichia coli for five, Proteus for three, Enterobacter cloacae for two, and Klebsiella, Enterobacter aerogenes, and Enterobacter vulnaris for one each. Two individuals with Gram-negative meningitis were admitted 72 to 96 hours after hospital discharge following pump replacement. Both patients had rapid deterioration requiring transfer to the pediatric intensive care unit, and developed coagulopathy and decrease in responsiveness. Both have improved and have elected not to replace the ITB pump. In Gram-negative infections in ITB therapy, the progression of signs and symptoms can be swift and devastating. Identification of the infectious agent in such cases is imperative; these infections can quickly become life threatening. PMID- 16700936 TI - The use of tiagabine in pediatric spasticity management. AB - Tiagabine, developed as an anti-epileptic medication, has the potential to reduce spasticity. The purpose of the present study was to assess the effectiveness of tiagabine in decreasing spasticity and improving the functional abilities of children with spastic cerebral palsy (CP). Nine children (seven females, two males) with CP (six spastic quadriplegia, three moderate to severe spastic diplegia) were treated with tiagabine for a mean of 7.2 months. Median age was 4y 5mo (range 3y 2mo-10y). All children were non-ambulatory. According to the Gross Motor Function Classification System, six were Level IV and three were Level V. Only one child showed a median decrease >or=1.0 grade on the modified Ashworth scale in upper extremities, lower extremities, and overall. Another child had significant improvement in the Pediatric Evaluation of Disability Inventory Self care score and improved feeding. None of the participants was found to have a significant improvement in motor function or a decrease in the number of motions (passive range of motion and muscle length test) that were limited. Reduction of nocturnal awakenings from painful spasms was reported in one child. Eight of the nine children experienced adverse side-effects during treatment. Although tiagabine was not found to be effective in decreasing children's spasticity or improving their function, its potential use in the relief of painful spasms associated with neurological conditions in the pediatric population warrants further investigation. PMID- 16700937 TI - Abnormal physiological responses to touch among children with persistent feeding difficulties. AB - Pediatric feeding/swallowing specialists commonly observe infants and toddlers who present with gagging or significant behavior state changes as a result of touch to oral and body regions. To date, this behavior has not been fully characterized or documented experimentally. This paper describes an exploratory study aimed at filling in these lacunae and providing a possible explanation. The study compares the responses to firm tactile pressure of 40 children between the ages of 3 and 18 months. The experimental group consisted of 20 children (12 males, 8 females) with heterogeneous medical diagnoses, at least 2 weeks of alternative feedings during the first 3 months of life, and a common history of persistent feeding difficulties. The comparison group consisted of 20 children with an unremarkable medical history matched by age and sex to the experimental group. Firm, tactile pressure was applied in a predetermined sequence following dermatome regions from legs to mouth. Response was recorded in terms of presence/absence of gagging and/or behavior state changes. Fifteen of the 20 children in the experimental group showed gagging (14) or inconsolable crying (1) in response to touch. None of the matched comparison group demonstrated either gagging or behavior state changes. The experimental and comparison groups showed significant differences in response to tactile input. Further, the source of these abnormal responses must be sought in similarity of experience across heterogeneous medical diagnoses and interventions. One possible explanation is a history of oral deprivation due to alternative feedings. PMID- 16700938 TI - Sex differences in prenatal oral-motor function and development. AB - The aim of this study was to investigate sex-related differences in the prenatal development of early oral, lingual, pharyngeal, and laryngeal motor activities. Sonographic images of oral-upper airway regions were observed in 85 healthy fetuses (43 males, 42 females; mean gestational age 24wks 3d [SD 0.69]; range 15 38wks). Biometric data on morphologic development and associated motor patterns were compared across second and third trimesters. Results showed that while males and females demonstrated statistically similar patterns of general physical growth (p>0.05), significant differences in development of specific lingual and pharyngeal structures were present (p<0.05). Significant differences were found for laryngeal and pharyngeal motor activity, and oral-lingual movements (p<0.05). Complex oral-motor and upper airway skills emerged earlier in females, suggesting a sex-specific trajectory of motor development. It was concluded that differential patterns of prenatal motor development may be important in defining sex-specific indices of oral skill maturation. PMID- 16700939 TI - Biobehavioural reactivity to pain in preterm infants: a marker of neuromotor development. AB - In this preliminary study, it was examined whether capacity to react to external stress (acute pain) during neonatal intensive care predicts later neuromotor development at 4 and 8 months corrected chronological age (CCA) in high-risk preterm infants. Behavioural and cardiac reactivity to blood collection at 32 weeks postconceptional age (PCA) were recorded in addition to developmental outcomes at 4 and 8 months CCA in 35 preterm infants (17 males, 18 females) born 38 years; younger, < or = 38 years). A total of 193 oocytes from 29 patients were studied. The younger group contained 100 oocytes and the older group 93 oocytes. The ZP was significantly thicker in metaphase II oocytes in the older group compared with the younger group (mean +/- SD: 24.1 +/- 2.5 microm vs 23.1 +/- 3.3 microm; p = 0.01) but ZP density was equal (2.8 +/- 0.7 nm). By day 2 of culture, embryos from the two groups had similar ZP thickness (22.2 +/- 2.2 microm vs 21.7 +/- 1.6 microm; p = 0.28) and density (2.9 +/- 0.7 nm vs 2.8 +/- 0.8 nm; p = 0.57). For the embryos cultured to blastocyst (older: n = 20; younger: n = 18) ZP thickness was similar in the two groups (19.2 +/- 2.7 microm vs 19.1 +/- 5.0 microm; p = 0.8) but thinner than on day 2. The older group had significantly denser ZP than the younger group (4.2 +/ 0.5 nm vs 3.3 +/- 1.0 nm, p < 0.01). Blastocysts from both groups had significantly denser ZP than their corresponding day 2 embryos (older: 4.2 +/- 0.5 nm vs 2.9 +/- 0.7 nm, p < 0.001; younger: 3.3 +/- 1.0 nm vs 2.8 +/- 0.8 nm, p = 0.013). It is concluded that there is little relationship between ZP thickness and its density as measured by polarized light microscopy. While ZP thickness decreases with extended embryo culturing, the density of the ZP increases. ZP density increases in both age groups with extended culture and, interestingly, more in embryos from older compared with younger women. PMID- 16700975 TI - Contribution of spermatozoal centrosomes to the microtubule-organizing centre in Antarctic minke whale ( Balaenoptera bonaerensis ). AB - Using an interspecies microinsemination assay with bovine oocytes, it was examined whether centrosomes of Antarctic minke whale spermatozoa function as the microtubule-organizing centre (MTOC). Bull and rat spermatozoa were used as positive and negative controls, respectively. Vitrified-warmed bovine mature oocytes were subjected to immunostaining against alpha-tubulin 4-6 h after intracytoplasmic injection (ICSI) of 5 mM dithiothreitol-treated spermatozoa. Aster formation occurred from whale spermatozoa (33%) and bull spermatozoa (33%), but very little from rat spermatozoa (3%). Activation treatment for the microinseminated oocytes with 7% ethanol + 2 mM 6-dimethylaminopurine resulted in a similar proportion of oocytes forming a whale sperm aster (35% vs 27% in the non-treated group; 4 h after ICSI) but a significantly larger aster (ratio of aster diameter to oocyte diameter, 0.57 vs 0.30 in the non-treated group). These results indicate that the centrosome introduced into bovine oocytes by whale spermatozoa contributes to the MTOC and that assembly of the microtubule network is promoted by oocyte activation. PMID- 16700976 TI - Relationship between bovine oocyte morphology and in vitro developmental potential. AB - We investigated the relationship between the morphology of oocytes collected from small antral follicles and their developmental capacity. Immature oocytes were classified into seven groups and cultured in vitro for maturation (IVM), fertilization (IVF) and development to blastocysts (IVC). After IVF, sperm penetration and normal fertilization rates were higher in the oocytes whose cytoplasm appeared brown. The rate of polyspermy was highest in the oocytes whose cytoplasm was black. After IVC, the rates of cleavage and of development to the blastocyst stage were also higher in the brown oocytes. Although the oocytes with dark clusters in a pale cytoplasm showed lower cleavage rates, cleaved zygotes had high developmental rates the same as the oocytes with a brown cytoplasm. Transmission electron microscopy showed that the oocytes with a pale or black cytoplasm had organelles arranged differently from other oocytes before IVM. Most of the oocytes with a brown and homogeneous cytoplasm or small diameter had the characteristics of immature cytoplasm (large clusters of cortical granules) even after IVM. On the other hand, the brown oocytes with a dark zone at the periphery or with dark clusters showed the same arrangement of organelles as in vivo matured oocytes. The oocytes with a pale or black cytoplasm appeared to be degenerating and/or ageing. In conclusion, a dark ooplasm indicates an accumulation of lipids and good developmental potential, while a light-coloured ooplasm indicates a low density of organelles and poor developmental potential. A black ooplasm indicates ageing and low developmental potential. PMID- 16700977 TI - A potential relationship between the acrosome response characteristics of bovine spermatozoa and their in vitro fertilizing ability. AB - The aim of the work was to study a potential relationship between acrosome response characteristics of bovine spermatozoa and their ability to fertilize oocytes and produce in vitro embryos. Sperm of artificial insemination bulls with a high rate (22.0 +/- 4.1%, group A, n = 7) or a low rate (10.3 +/- 4.1%, group B, n = 8) of embryos were used. For acrosome assessment, motile spermatozoa from a Percoll gradient were incubated with or without heparin and examined by the fix vital sperm assay (FVSA). The differences between the heparin-treated (H+) and the non-treated (H-) spermatozoa were significant (p < 0.01) in all bulls at all tested intervals. According to the kinetics of the heparin response, the bulls fell into three categories: fast (FR, n = 7), moderate (MR, n = 5) or slow (SR, n = 3) acrosome responses (p < 0.01). Five MR bulls were found in group A in comparison with two MR bulls in group B (57.1 vs 12.5%; p < 0.05). Intensity of the acrosome response (response index) was significantly higher in bull group A compared with bull group B (7.0 vs 4.6, p < 0.01). A positive correlation was recorded between response index and embryo rate (r = 0.668, p < 0.01). In conclusion (a) the kinetics of spermatozoa response to heparin may be important for in vitro fertilization, bulls with a moderate response appearing to be most suitable for embryo production; (b) greater spermatozoa response to heparin was related to more effective embryo production. PMID- 16700978 TI - Annealing control primer system identifies differentially expressed genes in blastocyst-stage porcine parthenotes. AB - There is very little information available on stage-specific gene expression during early embryo development, particularly in the pig. Here, we accurately identified the genes that are specifically or prominently expressed in parthenogenetic porcine blastocysts as compared with 2-cell stage embryos. We accomplished this by using a PCR technology regulated by annealing control primers (ACPs). By utilizing 120 ACPs, a total of 46 expressed sequence tags (ESTs) of genes that are differentially expressed in blastocysts as compared with 2-cell stage embryos were cloned and sequenced. The cloned genes or ESTs all exhibited significant sequence similarity with known genes or ESTs of other species. Of the known genes, six genes [renin-binding protein (RNBP), BMDP, solute carrier family 25 (SLC25A6), MTHFD1, TRK-fused gene (TFG), spermidine synthase (SRM)] were selected and their stage-specific expression levels in porcine parthenotes were determined by real-time quantitative polymerase chain reaction at the 1-, 2-, 4-cell, morula and blastocyst stages. While RNBP, BMDP, SLC25A6, MTGFD1 and SRM were highly expressed only at the blastocyst stage, TFG was highly expressed at the 1-cell stage, then declined after genomic activation, high levels of expression being again detected at the morula and blastocyst stages. This analysis suggests that the ACP system is an effective tool for use in the identification of stage-specific genes in small numbers of porcine parthenotes. Examination of the genes differentially expressed in the blastocyst, which we have identified here, will provide insight into the molecular basis of preimplantation development. PMID- 16700979 TI - Differential effects of culture and nuclear transfer on relative transcript levels of genes with key roles during preimplantation. AB - It is well known that the preimplantation culture environment to which embryos are exposed influences the expression of developmentally important genes. Recently, it has been reported that MEMalpha, a culture medium commonly used for somatic cells, allows high rates of preimplantation development and development to term of mouse somatic cell nuclear transfer (SCNT) embryos. The objective of this study was to compare the differential effects of this medium and of the nuclear transfer procedure on the relative mRNA abundance of several genes with key roles during preimplantation. The relative mRNA levels of nine genes (Glut 1, Glut 5, G6PDH, Bax, Survivin, Gpx 1, Oct4, mTert and IGF2bp1) were quantified at blastocyst stage on cumulus cell cloned embryos cultured in MEMalpha, as well as on in vivo cultured and MEMalpha cultured controls. Only three of the nine transcripts analysed (Glut 5, Gpx 1 and Igf2bp1) were significantly down regulated at blastocyst stage in in vitro produced controls. However, most genes analysed in our MEMalpha cultured cloned embryos showed altered transcription levels. Interestingly, between cloned and in vitro produced controls only the transcription levels measured for Glut 1 were significantly different. This result suggests that Glut 1 may be a good marker for embryo quality after cumulus cell nuclear transfer. PMID- 16700983 TI - [Problems and thoughts on the relationship between experimental study and clinical study of senile cerebral ischemia]. PMID- 16700984 TI - [Study on safety of apnea test in clinical determination of brain death]. AB - OBJECTIVE: To determine the occurrence of severe complications such as hypotension, pulmonary artery hypertension as well as hypercapnia during apnea test in the affirmation of brain death and to investigate the possible effective prophylactic interventions. METHODS: Conventional apnea test was performed in 15 clinically suspected brain death patients. Stable circulation was achieved by adjusting preload only (n=4) or combined with titrating norepinephrine (NE, n=11). Blood gas was respectively analyzed before apnea test, 10 minutes after 100% fraction of oxygen (FiO(2)) ventilation, at each 2-minute interval after disconnecting ventilator and 5 minutes after reventilation. Hemodynamic parameters and dosage of NE were recorded at the same time points. Plasma concentration of lactate was measured before and at the end of apnea test. RESULTS: Spontaneous breath occurred in 1 case among 15 suspected brain death patients. Partial pressure of carbon dioxide (PaCO(2)) reached higher than 60 mm Hg (1 mm Hg=0.133 kPa) within 8 minutes in positive apnea test patients (P<0.01). pH significantly decreased (P<0.05), but partial pressure of oxygen (PaO(2)) maintained higher than 100 mm Hg during the test. Heart rate (HR) and mean artery pressure (MAP) slightly lowered (P>0.05), but pulmonary artery pressure (PAP) markedly elevated (P<0.05) at the end of the test in comparison with their base lines. On the other hand, HR and MAP increased in the negative apnea test case after ventilator disconnection. Severe arrhythmia events did not occur in all the cases. There was no change in the dosage of NE infusion, the range of which was 0.10-0.60 microg.kg(-1).min(-1) with the mean level of (0.23+/-0.17) microg.kg( 1).min(-1). The trend of HR, MAP, PAP and pulmonary arterial wedge pressure (PAWP) alterations was the same in patients no matter whether or not NE was used. HR, MAP and PAWP lowered, while PAP enhanced. Plasma lactate level was not significantly altered at the end of the test compared with the base line (from (1.41+/-0.05) mmol/L to (1.47+/-0.07) mmol/L). CONCLUSION: Adequate oxygenation could be maintained during conventional apnea test. The risk of inducing severe hypotension is low in non brain death patients. Based on adequate preload, low dose of NE infusion could prevent patients with high risk circulation instability from severe hypotension. PMID- 16700986 TI - [Relationship between methylenetrahydrofolate reductase gene and ischemic stroke]. AB - OBJECTIVE: To study the relationship between methylenetrahydrofolate reductase (MTHFR) gene and ischemic stroke. METHODS: Four hundred and fifty four ischemic stroke patients were enrolled in the study. They were divided into large artery atherosclerosis (LAA), cardioembolism (CE), small artery occlusion (SAA), stroke of other determined etiology (SOE) and stroke of undetermined etiology (SUE) according to TOAST (Trail of ORG 10172 in Acute Stroke Treatment) criteria; and they were divided into mild, moderate and severe types ischemic stroke according to their scores of neurologic impairment. Three hundred and thirty four subjects, in whom hypertension, coronary heart disease, cerebral vascular disease, diabetes mellitus, cancer, renal failure etc. were excluded, served as controls in the study. Their C677T polymorphisms of MTHFR gene were determined with polymerase chain reaction (PCR) and denaturing high performance liquid chromatography (DHPLC), and their risk factors of ischemic stroke were recorded at the same time. RESULTS: The frequencies of CC, CT and TT genotype in ischemic stroke were 51.8%, 40.5% and 7.7%, respectively, and they were 56.9%, 38.3% and 4.8% respectively in controls. TT genotype and T allele were associated with LAA and CE, moderate type and severe type of ischemic stroke. The frequencies of TT genotype and T allele in ischemic stroke patients were significantly higher in those with smoking, alcohol abuse or diabetes mellitus than those in controls (all P<0.10), but CC genotype and C allele were significantly lower in them than those in controls (all P<0.05). On the other hand, all of genotypes and alleles in ischemic stroke patients with no history of smoking, alcohol abuse or diabetes mellitus were not significantly different from those in controls. CONCLUSION: TT genotype and T allele are risk factors for ischemic stroke. It exists interactions between smoking, alcohol abuse, diabetes mellitus and MTHFR gene in the pathogenesis of ischemic stroke. PMID- 16700987 TI - [Changes in electroencephalograph and somatosensory evoked potential and their relationship with neuron apoptosis in rat after ischemic insult to brain]. AB - OBJECTIVE: To study the changes in electroencephalograph (EEG) and somatosensory evoked potential (SEP) and their relationship with neuron apoptosis in rat after ischemic insult to the brain. METHODS: Thirty-five SD rats were randomly divided into normal, sham operated and 3, 12, 24, 48, 72 hours after ischemia/reperfusion (I/R) groups with 5 rats in each group. The ischemia of brain was produced by clamping 4 vessels to the brain for various periods of time. Changes in EEG and SEP were recorded at different time after I/R, and the amounts of apoptotic neurons in hippocampus and cortex after I/R were assessed with terminal deoxynucleotidyl transferase mediated dUTP biotin nick end labeling (TUNEL) and acridine orange ethidium bromide (AO/EB) fluorescence examination techniques. RESULTS: Compared with sham operated group, EEG amplitude decreased significantly (all P<0.05), and the proportion of Delta wave increased significantly after ischemia of the brain (all P<0.05). The latent period of P1 wave crest extended markedly (all P<0.05), and P1-N1 amplitude decreased significantly after I/R (all P<0.05). EEG and SEP changes were correlated with the apoptosis and loss of neurons, which started in the hippocampus and extended to frontal cortex and parietal cortex. CONCLUSION: The combined analysis of EEG and SEP can reflect the process of neuron apoptosis, which is helpful for the diagnosis and evaluation of prognosis of patients suffering from cerebral ischemia. PMID- 16700989 TI - [Establishment and evaluation of reproduction of middle cerebral artery occlusion to produce cerebral ischemia with autologous blood clot and nylon thread in rat]. AB - OBJECTIVE: To reproduce cerebral ischemia using middle cerebral artery, occlusion (MCAO) method with autologous blood clot and nylon thread in rat. METHODS: Rats were randomly divided into sham operation group (n=10), suture occlusion group (n=12), thrombus suture occlusion model group (n=12) in which both autologous blood clot and nylon thread were used (thrombus occluded group). Cerebral ischemia as a result of MCAO in suture occlusion group was replicated by insertion of nylon thread. In thrombus occluded group, a polyethylene catheter containing a nylon thread was inserted into the middle carotid artery via external carotid artery, and the nylon thread was pulled out to an appropriate length to allow blood to coagulate. Fifteen minutes later, thrombin was injected, and the nylon thread was re-inserted to complete MCAO. Specimens were taken after ischemia for 6 hours. Neurological symptoms were observed, brain water ratio and the cerebral infarction size were measured, and the pathological changes in the brain tissue were observed. RESULTS: Compared with sham operation group, the evaluation score of neurological symptoms was significantly higher, and water content and cerebral infarction size were larger in suture occlusion group and thrombus occluded group (all P<0.01); the pathological changes of the brain were also more marked. Compared with suture occlusion group, thrombus occlusion group showed no differences in evaluation score, water content, cerebral infarction size and pathological changes of the brain (all >0.05). CONCLUSION: In rats, occlusion of middle cerebral artery with autologous blood clot and inserted nylon thread can result in the functional lesion, edema and the pathological changes in the brain. The MCAO reproduced by this method could produce thrombotic obstruction of the middle cerebral artery for the experimental study of cerebral ischemic/reperfusion injury, and it could prevent hemorrhage in the process of replication of animal model and in the course of thrombolysis therapy in the clinic. The model has the advantages of precision of the occlusion site, production of constant cerebral infarction size, easier performance and good repeatability. PMID- 16700990 TI - [Effects of mild hypothermia on expression of N-methyl-D-aspartate receptor-1 in hippocampus neurons after cardiopulmonary resuscitation in rats]. AB - OBJECTIVE: To explore the effects of mild hypothermia on expression of N-methyl-D aspartate receptor-1 (NMDAR1) in hippocampus neurons after cardiopulmonary resuscitation (CPR) in rats. METHODS: Twenty-four male SD rats were randomly divided into normal control group, normal temperature group, and mild hypothermia group, with 8 rats in each group. The cerebral edema model after CPR was replicated by the sealed bottle method in rats in both normal temperature group and mild hypothermia group. The rats in the mild hypothermia group were treated with mild hypothermia after the model was established. The change in expression of NMDAR1 in hippocampus neurons in rat was determined with semi-quantitative reverse transcription polymerase chain reaction (RT-PCR), and pathologic changes in brain tissue were observed in both groups. RESULTS: The cerebral edema was ameliorated, NMDAR1 mRNA and protein in cerebral hippocampus neurons were significantly lower in hypothermia group than control group with significant difference (NMDAR1 mRNA: 80.48+/-0.03 vs. 80.64+/-0.18, P<0.05 ). CONCLUSION: Mild hypothermia can down regulate the expression of NMDAR1 mRNA and protein level, lower positive ion concentration and thus decrease cerebral edema, so mild hypothermia can serve as a treatment measure for cerebral edema after CPR. PMID- 16700991 TI - [Changes in inflammatory cascade response and its implication during cerebral ischemia/reperfusion in aged rats]. AB - OBJECTIVE: To investigate the changes in expression of tumor necrosis factor alpha(TNF-alpha), vascular cellular adhesive molecule-1 (VCAM-1), intercellular adhesion molecular-1 (ICAM-1) and its mRNA expression, and the pathogenetic mechanism of cerebral ischemia/reperfusion(I/R) in elderly. METHODS: Thirty six male young SD rats (5-6 months old) and 36 male aged SD rats (20-21 months old) were randomly divided into young sham operated group, young model group, aged sham operated group and aged model group, respectively. The two model groups were randomly divided into ischemia 3 hours (I 3 h), I/R 1, 3, 6, 12 days groups. There were 6 rats in each group. Focal cerebral I/R model was replicated with middle cerebral artery occlusion (MCAO). The changes of the nervous dysfunction score, the water content of cerebral constitution and the expression of TNF alpha, VCAM-1, ICAM-1 and ICAM-1 mRNA were observed at each time point. RESULTS: The expression of TNF-alpha in the aged sham operated group were higher than that of the young sham operated group. The nervous dysfunction score (I 3 h and I/R 1 12 d), the water content of cerebral constitution (I/R 1-6 d), the expression of TNF-alpha (I 3 h and I/R 1-6 d), VCAM-1 (I 3 h and I/R 1 d), ICAM-1 (I 3 h and I/R 1-6 d) and ICAM-1 mRNA (I 3 h and I/R 1-12 d) in young model group and aged model group were higher than those of the young sham operated group and the aged sham operated group respectively. The nervous dysfunction score (I 3 h, I/R 6 d), the expression of TNF-alpha (I/R 1, 3 d), VCAM-1 (I/R 3, 6 d), ICAM-1 (I 3 h, I/R 1 d) and ICAM-1 mRNA (I/R 1-6 d) in aged model group were higher than that of young model group. CONCLUSION: The cerebral I/R injury is correlated with the up regulation of TNF-alpha, VCAM-1,ICAM-1 and its mRNA expression. The cerebral I/R injury of aged rats is more serious than that of young rats which might be associated with up regulation of TNF-alpha, adhesion molecules expression with aging. PMID- 16700992 TI - [Comparison of curative effect and analysis of influencing factors in patients with basal ganglion hemorrhage treated with traditional craniotomy and mini invasive operation]. AB - OBJECTIVE: To compare the curative effect of traditional craniotomy and mini invasive operation on basal ganglion hemorrhage, and analyze influencing factors related to the differences. METHODS: Fifty-five cases with basal ganglion hemorrhage according with the enrollment criterion of standardized treatment of hemorrhagic stroke were analyzed. Twenty-eight cases were treated with mini invasive operation (mini-invasive group) and 27 cases with traditional craniotomy (craniotomical group). The evaluation indexes included postoperative complications, consciousness in 3 days and 2 weeks after operation, and activity of daily life (ADL) in 3 months after operation. SPSS 10.0 software was used for statistical analysis. RESULTS: Three days after operation, 19 cases and 10 cases of abnormal consciousness were in craniotomical group and mini-invasive group, respectively (P<0.01). Seventeen cases had pulmonary infection and/or organ dysfunction in craniotomical group, and 11 cases in mini-invasive group (P<0.05). Three months after operation, 5 cases were in vegetable status and 3 died in craniotomical group, accounting for 29.6%, while 3 cases were in vegetable status and 2 died in mini-invasive group, accounting for 17.8% (P<0.05). No difference in ADL was found in the two groups with an exception of cases of vegetable status and death (>0.05). CONCLUSION: mini-invasive operation makes the abnormal consciousness recover earlier, reduces complications, vegetable status and mortality and has no obvious effect on recovery of neurological function. PMID- 16700993 TI - [Changes in cerebral oxygen metabolism and significance in patients with severe head injury]. AB - OBJECTIVE: To investigate the changes in cerebral oxygen metabolism following head injury and their relationship with intracranial pressure (ICP), cerebral perfusion pressure (CPP), and Glasgow coma scale (GCS). METHODS: Forty-five patients after surgery for serious head injury were studied and divided into different groups according to ICP, CPP and GCS, 10 patients without head injury were enrolled as controls. GCS, ICP, CPP, blood gas analysis, jugular bulb for gas analysis, jugular bulb blood oxygen saturation (SjvO(2)), cerebral arterial venous oxygen content difference (AVDO(2)) were accessed and scored, and cerebral extraction of oxygen (CEO(2)) were calculated in all the patients. The relationships of these parameters with ICP, CPP and GCS score were analysed. RESULTS: On the 1 st postinjury day SjvO(2) decreased whereas CEO(2) and AVDO(2) increased in all patients, with significant differences compared with the controls (all P<0.05). During the 2nd-4th postoperative days, SjvO(2) gradually increased with CEO(2) and AVDO(2) decreased in patients with mild or moderate head injury of increased in ICP or lowered in CPP group, the changes in the above parameters were slower, with significant difference compared with patients with increase in ICP, and mild or moderate lowering of CPP (all P<0.05). There were significant differences in SjvO(2), CEO(2), and AVDO(2) between the group of GCS<5 and GCS>6-8 during the 2nd-5th days (all P<0.05). ICP was negatively correlated with SjvO(2) (r=-0.8652, P<0.01) and positively correlated with CEO(2) (r=0.4172, P<0.05) and AVDO(2) (r=0.4771, P<0.05). CPP was positively correlated with SjvO(2) (r=0.8830, P<0.01) and negatively correlated with CEO(2) (r=-0.6724, P<0.05) and AVDO(2) (r=-0.8350, P<0.01). GCS score was positively correlated with SjvO(2) (r=0.8230, P<0.01) and CEO(2) (r=0.8010, P<0.001), but there was no correlation between GCS score and AVDO(2) (r=2.6310, P=0.677). CONCLUSION: In patients with serious head injury, if the increase in ICP is mild or moderate, there are cerebral hypoxia and ischemia during the first 24 hours after injury. During the 2 nd-4 th postinjury days, the brain becomes hyperemic and hyperoxygenated. Marked increase in ICP results in persistent cerebral hypoxia and ischemia. ICP, CPP and GCS score are main factors affecting cerebral oxygen metabolism. PMID- 16700995 TI - [Study of relationship between inflammatory response and apoptosis in perihematoma region in patients with intracerebral hemorrhage]. AB - OBJECTIVE: To investigate the relationship between inflammatory response and cell apoptosis in the perihematoma region in patients with intracerebral hemorrhage (ICH). METHODS: Surgical specimens were obtained from the area 1 cm adjacent to the hematoma. Thirty patients with ICH were divided into five groups: 6, 7, 5, 6, 6 patients in surgery<6 hours, 6-12 hours, 12-24 hours, 24-72 hours and >72 hours groups after the onset, respectively. The control group specimens were obtained from the brain tissues distant to the hematoma in the process of craniotomy in the patients of two former groups. Sections were stained with hematoxylin and eosin (HE) for the examination of pathological changes. Immunohistochemistry, terminal deoxynucleotidyl transferase mediated dUTP biotin nick end labeling (TUNEL) and reverse transcription-polymerase chain reaction (RT-PCR) were applied to determine apoptosis cells, Bax and Bcl-x protein and mRNA. RESULTS: The tissues from perihematoma region were almost normal in control group and <6 hours group. They were slightly damaged in 6-12 hours group, became worse in 12-24 hours group and most severe in 24-48 hours group, and they became better latter and were similar to the control group on 8th day. Infiltration of neutrophils, macrophages and lymphocyte appeared gradually at 6-12 hours, and became much more prominent at 12-24 hours (all P<0.01). The reactive gliosis began to appear at 24 72 hours, and enhanced after 72 hours (all P<0.01). The expression of the apoptosis and Bax protein increased gradually after 6 hours, reaching the peak at 12-24 hours (P<0.05 or P<0.01), and decreased gradually later. The changes in the levels of Bax mRNA were similar to that of the result of immunohistochemistry. Although the expression of Bcl-x protein and mRNA seemed to be increased at 12-72 hours, there was no significant difference between groups (P>0.05). The correlation analysis showed that the infiltration of neutrophils, macrophages and lymphocyte was positively correlated to the TUNEL positive cells and expression of Bax protein and mRNA (P<0.05 or P<0.01), and showed no correlation to Bcl-x protein and mRNA (all >0.05). CONCLUSION: There is a close relationship between inflammatory response and apoptosis and tissue damage in the perihematoma area in ICH. PMID- 16700996 TI - [Effect of head hypothermia on serum inflammatory cytokines levels in patients with hypertensive intracerebral hemorrhage]. AB - OBJECTIVE: To study changes in interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) levels in hypertensive intracerebral hemorrhage (HIH) and observe the potential effect of head hypothermia. METHODS: One hundred and twenty four cases of HIH were divided into head hypothermia treatment group (n=63) and control group (n=61) according to treatment with or without head hypothermia. The serum IL-6 and TNF-alpha level in the two groups were determined on the 2nd and 8th days with enzyme linked immunoadsorbent assay (ELISA) and radioimmunoassay, and compared to those of the healthy volunteers (n=40). In addition, the life quality scores of the two groups were compared. RESULTS: The serum IL-6 and TNF alpha levels in both head hypothermia treatment group and control group were higher on the 2nd day after hospitalization compared with the healthy controls (all P<0.01), and there was no significant difference in serum IL-6 and TNF-alpha levels on the 2nd day between the head hypothermia treatment group and control group (both >0.05). Serum IL-6 and TNF-alpha levels in the head hypothermia treatment group on the 8th day after hyperthermia were significantly lower than those on the 2nd day after hospitalization (both P<0.05), while the changes of control group was not significant (both >0.05), and significant difference in serum IL-6 and TNF-alpha was found between head hypothermia treatment group and control group on day 8 (both P<0.01). The prognosis quality of life in head hypothermia treatment group was markedly higher than the control group (P<0.05). CONCLUSION: The levels of serum IL-6 and TNF-alpha were increased significantly during early stage of HIH. Head hypothermia can effectively reduce the serum IL-6 and TNF-alpha levels of early HIH patients, and improve the prognosis quality of life as well. PMID- 16700998 TI - [Study on plasma antibodies against heat shock protein 70 in the hypertensive patients with cerebral infarction]. AB - OBJECTIVE: To determine the level of heat shock protein 70 (HSP70) antibody in the hypertensive patients with cerebral infarction and explore the role of plasma antibody against HSP70 in the pathogenesis of cerebral infarction and its relationship to the brain tissue damage. METHODS: One hundred patients with CT or MRI confirmed cerebral infarction were enrolled in the study, in whom 64 were male, and 36 female. Among them, 67 patients were having hypertension and 33 with normal blood pressure. One hundred healthy individuals, including 58 male and 42 female, served as the control group. The level of plasma HSP70 antibody was determined with Western blot-enzyme linked immunoadsorbent assay (ELISA) technique. RESULTS: Patients with cerebral infarction had a significantly higher incidence (69%) of antibodies to HSP70 compared to the low incidence (13%) of antibodies to HSP70 in the controls. Patients with hypertension had a high incidence (57%) of antibodies to HSP70 compared to the low incidence (12%) of antibodies to HSP70 in patients with normal blood pressure in cerebral infarction group. Patients with hypertension in the control group had a higher incidence (11%) of antibodies to HSP70 compared to the low incidence (2%) of antibodies to HSP70 in the same group. CONCLUSION: There is a significant difference in the level of antibodies to HSP70 between patients with cerebral infarction and controls. Elevation in the level of antibodies to HSP70 may reflect the degree of brain tissue damage. PMID- 16700999 TI - [Research on association between atherosclerotic brain infarction and human leukocyte antigen DR gene]. AB - OBJECTIVE: To evaluate the relationship between atherosclerotic brain infarction (ABI) and human leukocyte antigen DR (HLA-DR) gene at molecular level. METHODS: By using polymerase chain reaction sequence specific probe (PCR-SSP), gene special position of the HLA-II DR alleles were determined in 31 patients and 30 healthy individuals as controls, all of them were inhabitants of Northeastern China. RESULTS: The HLA-DR B1*0301 gene frequency in the group of ABI was obviously higher than that of control group (RR=5.6842, P<0.05). CONCLUSION: The HLA-DR B1*0301 gene may be the susceptible gene of ABI in Northeastern China, or it may associate with other genes to produce the disease. PMID- 16701000 TI - [Expression of matrix metalloproteinases and their inhibitors in fetal skin and their biological significance]. AB - OBJECTIVE: To investigate the expression characteristics of matrix metalloproteinases (MMP-2, MMP-3, MMP-9) and tissue inhibitor of metalloproteinase (TIMP-1, TIMP-2) in developing fetal skin and their potential biological significance. METHODS: Skin of 24 cases of fetuses with different gestational age (12-40 weeks) were obtained, embedded with paraffin wax, and sectioned. Immunohistochemistry and pathological methods were used to detect the expression intensity and distribution of MMP-2, MMP-3, MMP-9, TIMP-1 and TIMP-2. RESULTS: Positive immunohistochemical signals of MMPs and TIMPs could be found in fetal skin at different gestational periods. These proteins mainly located in the cytoplasm of epidermal cells, fibroblasts, epithelial cells of hair follicles and sweat glands and vascular endothelial cells. In earlier gestational fetal skin (12-18 weeks), the expression levels of MMP-2, MMP-3 and MMP-9 were high. Along with the advancement in gestational age, the positive rates of these three proteins in skin were lowered, and in later gestational fetal skin (27-40 weeks) the expression rates were significantly decreased compared with those in earlier gestational fetal skin (all P<0.05). On the contrary, protein expression levels of TIMP-1 and TIMP-2 were apparently lower in earlier versus later gestational skin (both P<0.05). CONCLUSION: MMP-2, MMP-3, MMP-9, TIMP-1 and TIMP-2 might be involved in the skin development and maintenance of cutaneous structure and function. Higher expression of MMP-2, MMP-3 and MMP-9 and lower protein levels of TIMP-1 and TIMP-2 may provide an antiscarring signal in healing of wound during early periods of gestation. PMID- 16701009 TI - Restoration of shoulder abduction by transfer of the spinal accessory nerve to suprascapular nerve through dorsal approach: a clinical study. AB - BACKGROUND: In recent years, transfer of the spinal accessory nerve to suprascapular nerve has become a routine procedure for restoration of shoulder abduction. However, the operation via the traditional supraclavicular anterior approach often leads to partial denervation of the trapezius muscle. The purpose of the study was to introduce transfer of the spinal accessory nerve through dorsal approach, using distal branch of the spinal accessory nerve, to repair the suprascapular nerve for restoration of shoulder abduction, and to observe its therapeutic effect. METHODS: From January to October 2003, a total of 11 patients with a brachial plexus injury and an intact or nearly intact spinal accessory nerve were treated by transferring the spinal accessory nerve to the suprascapular nerve through dorsal approach. The patients were followed up for 18 to 26 months [mean (23.5 +/- 5.2) months] to evaluate their shoulder abduction and function of the trapezius muscle. The outcomes were compared with those of 26 patients treated with traditional anterior approach. And the data were analyzed by Student's t test using SPSS 10.5. RESULTS: In the 11 patients, the spinal accessory nerves were transferred to the suprascapular nerve through the dorsal approach successfully. Intact function of the upper trapezius was achieved in all of them. In the patients, the location of the two nerves was relatively stable at the level of superior margin of the scapula, the mean distance between them was (4.2 +/- 1.4) cm, both the nerves could be easily dissected and end-to-end anastomosed without any tension. During the follow-up, the first electrophysiological sign of recovery of the infraspinatus appeared at (6.8 +/- 2.7) months and the first sign of restoration of the shoulder abduction at (7.6 +/- 2.9) months after the operation, which were earlier than that after the traditional operation [(8.7 +/- 2.4) months and (9.9 +/- 2.8) months, respectively; P < 0.05]. The postoperative shoulder abduction was 62.8 degrees +/ 12.6 degrees after transfer of the spinal accessory nerve, better than that after the traditional (51.6 degrees +/- 15.7 degrees). All the 11 patients could extend and externally rotate the shoulder almost normally. CONCLUSIONS: The accessory nerve transfer through dorsal approach is a safe and reliable procedure for the treatment of brachial plexus injury. Its postoperative effect is confirmed, which is better than that of the traditional operation. PMID- 16701010 TI - Hypoxia-inducible factor-1alpha induces the epithelial-mesenchymal transition of human prostatecancer cells. AB - BACKGROUND: Hypoxia-inducible factor-1alpha (HIF-1alpha) is a transcriptional factor that could improve the stimulation of angiogenesis and the metabolic adaptation of tumor cells to hypoxia. A recent study showed that HIF-1alpha could induce colon cancer cells epithelial-mesenchymal transition (EMT). However, no evidence indicates a similar correlation in human prostate cancer cells. This study was designed to evaluate the effect of HIF-1alpha over-expression on the EMT in human prostate cancer cells. METHODS: We selected the appropriate cell line for HIF-1alpha induction from those EMT negative prostate cell lines through vimentin gene detection by RT-PCR. As the result, LNCaP cell line is the best one for further experiment. LNCaP cells were transfected with recombinant plasmid pcDNA3.1 (-)/HIF-1alpha and pcDNA3.1 (-) control vector by Lipofectamine 2000 system. The positive cell colonies were confirmed by indirect immunofluorescence labeling. Then Transwell polycarbonate filter was used to analyze the invasive potency. The expression of EMT associated proteins, E-cadherin and vimentin, was detected by Western blotting. RESULTS: Among four of the EMT negative cell lines, LNCaP was the only one expressed the vimentin gene but not the associated protein. The expression level of HIF-1alpha in LNCaP/HIF-1alpha was distinctly higher than that in LNCaP/pcDNA3.1 and LNCaP. The cell numbers of LNCaP/HIF 1alpha that penetrated through the Transwell filter were higher than that of LNCaP/pcDNA3.1 and LNCaP. Compared with the LNCaP/pcDNA3.1 and LNCaP cells, the expression of vimentin was up-regulated in LNCaP/HIF-1alpha, whereas the expression of E-cadherin was down-regulated. CONCLUSIONS: Over-expression of HIF 1alpha stimulates the invasion potency of human prostate carcinoma cells through EMT pathway. The expression of E-cadherin and vimentin, playing established roles in EMT, could be regulated by HIF-1alpha in human prostate cancer cell line. PMID- 16701011 TI - Impact of genetic variation of tumor necrosis factor-alpha on gestational hypertension. AB - BACKGROUND: The mechanisms responsible for the pathogeneses of gestational hypertension and preeclampsia are unclear. Tumor necrosis factor-alpha (TNF alpha) is a pro-inflammatory Th(1)-type cytokine. TNFA gene is located in the human leukocyte antigen (HLA) class III region of the major histocompatibility complex (MHC) on chromosome 6. The high TNF-alpha mRNA expression may be associated with the TNF2 (A) allele, which is the polymorphism of TNF-alpha at position -308 in promoter region. This study assessed whether the TNF2 (A) allele at position -308 plays a role in the alteration of blood pressure (BP) and urinary protein excretion during pregnancy. METHODS: The original prospective cohort study comprised 1623 pregnant women from January 2000 to October 2001. The G/A polymorphism was done by restriction fragment length polymorphism (RFLP) analysis with Nco I enzyme. RESULTS: The distributions of the G/A polymorphism of TNF-alpha in the promoter region at position -308 were wild-type 72.4% and variant 27.6%, respectively. The frequency of TNF2 (A) allele was approximately 0.15 for Caucasian pregnant women in the study. It was not significantly different in the distributions of genotypes and G/A allele frequencies among the three groups of pregnant women with gestational hypertension, preexisting hypertension and normal blood pressure (P > 0.05). The maternal blood pressure in the third trimester was significantly higher in the group of women possessing the TNF2 (A) allele compared to homozygous for the TNF1 (G) allele (systolic BP, P < 0.01 and diastolic BP, P < 0.05). The elevated blood pressure in the TNF2 (A) group was accompanied by higher urinary protein excretion in the third trimester (P < 0.05). The blood pressure and urinary protein excretion did not change apparently between the two groups in the first and second trimesters (P > 0.05). CONCLUSIONS: Maternal TNF2 (A) allele of TNF-alpha promoter region at position 308 could play a role in the alteration of blood pressures and/or enhancement of urinary protein excretion during pregnancy, and might play an important role in the development of both gestational hypertension and preeclampsia. PMID- 16701012 TI - Inhibition of microvascular endothelial cell apoptosis by angiopoietin-1 and the involvement of cytochrome C. AB - BACKGROUND: Angiopoietin-1 (Ang-1) is an endothelial-specific growth factor that can promote angiogenesis. Studies demonstrated that Ang-1 can inhibit apoptosis of umbilical endothelial cells, but so far little is known about its effects on apoptosis of microvascular endothelial cells. With the apoptotic model of murine cerebral-derived microvascular endothelial cells (bEnd.3) induced by serum-free culture, we attempted to clarify the molecular mechanism of bEnd.3 apoptosis, particularly its relation to cytochrome C (Cyt C). METHODS: The cultured microvascular endothelial cell strain, bEnd.3 cell, was employed. An apoptotic model of bEnd.3 was established by serum-free culture. Flow cytometry after Annexin labeling and PI staining were used to assess the apoptotic effects of Ang 1 on bEnd.3, and the expression of Bax/Bcl-2, caspase 8, caspase 3, and Cyt C were detected with Western blotting and ELISA. RESULTS: The apoptotic rate of bEnd.3 cells after stimulation with Ang-1 (100 ng/L) in serum-free medium was significantly higher than that in control group. Ang-1 inhibited early-stage apoptosis more than late-stage apoptosis provided by propidium iodide (PI) and AnnexinV double staining. The inhibition of Ang-1 on bEnd.3 cell apoptosis was strengthened with the increase in concentration (0 - 400 ng/ml). Ang-1 could decrease the expression of Bax, caspase3 and 8, and increase that of Bcl-2. The results of ELISA indicated that Ang-1 significantly decreased CytC content in cytoplasm and increase that in mitochondria. CONCLUSIONS: Ang-1 could inhibit bEnd.3 apoptosis induced by serum-free medium culture. The apoptosis was associated with decreased Bax expression, increased Bcl-2 expression, which result in Cyt C transferring from mitochondria to cytoplasm, and then caspases activation are reduced and cell apoptosis is suppressed. PMID- 16701013 TI - siRNA-mediated silencing of Cockayne Cyndrome group B gene potentiates radiation induced apoptosis and antiproliferative effect in HeLa cells. AB - BACKGROUND: Cockayne syndrome (CS) is a rare human genetic disorder characterized by increased UV sensitivity, developmental abnormalities and premature aging. Cells isolated from individuals with CS have a defect in transcription-coupled DNA repair. Despite the repair defect, there is no any increased risk of spontaneous or UV-induced cancer for CS individuals. The strategy of RNA interfering was used here to explore the potential radiosensitizing and anticancer activity of targeting CS group B (CSB) gene. METHODS: The vectors encoding CSB-specific siRNAs were constructed by inserting duplex siRNA encoding oligonucleotides into the plasmid P(silencer TM 3.1). The cell lines expressing the CSB-siRNA were generated from HeLa cells transfected with the above vectors. Colony-forming ability was used to assay cell survival. Cell cycle was analyzed by FACScan flow cytometry. The apoptosis was measured by detecting the accumulation of sub-G(1) population as well as by fluorescence staining assay. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to semi quantify mRNA expression. Protein level was detected by Western blotting analysis. RESULTS: Two constructs encoding CSB-specific siRNA were generated, both of them resulted in remarkable suppression on CSB expression in HeLa cells, and led to an increased sensitivity to (gamma-ray and UV light. siRNA-mediated silencing of CSB decreased cell proliferation rate, increased spontaneous apoptosis as well as the occurrence of UV- or cisplatin-induced apoptosis by 2 to 3.5 fold. A significant S phase blockage and a remarkable reduction of G(1) population were induced in control HeLa cells at 18 hours after being exposed to 10 J/m(2) of UV light. The S phase blockage was also observed in UV-irradiated CSB-siRNA transfected HeLa cells, but the extent of increased S phase population was lower than that in the UV-irradiated control cells. No or a relative weak reduction on G(1) phase population was observed in UV-irradiated CSB-siRNA transfected HeLa cells. In addition, siRNA-mediated silencing of CSB promoted the elimination of G(2)/M phase cells after UV light radiation. CONCLUSIONS: siRNA mediated silencing of CSB causes cells to proliferate more slowly, sensitize cells to genotoxicants, and modify UV radiation-induced cell cycle changes. siRNA mediated inactivation of CSB could be an attractive strategy for ameliorating cancer therapy, which can be fulfilled via the combination of gene therapy and sensitization of radiotherapy or chemotherapy. PMID- 16701014 TI - Role of T-cell receptor V beta 8.3 peptide vaccine in the prevention of experimental autoimmune uveoretinitis. AB - BACKGROUND: T-cell receptor (TCR) plays an important role in the development of autoimmune diseases. Recently, it was reported that immunization of animals with TCR peptide derived from the pathogenic cells could prevent autoimmune diseases. The aim of this study was to investigate whether vaccination with a synthetic peptide from the hypervariable region of TCR V(beta) 8.3, an experimental autoimmune uveoretinitis (EAU)-associated gene, was able to prevent the disease. METHODS: EAU was induced in Lewis rats by immunization with IRBP R16 peptide emulsified in complete Freund's adjuvant (CFA). The clinical and histological appearances were scored. Delayed type hypersensitivity (DTH) and lymphocyte proliferation were detected. Cytokine levels of aqueous humour, supernatants of cells from spleen and draining lymph nodes were measured by enzyme linked immunosorbent assay (ELISA). Gene expression of TCR V(beta) 8.3 on CD(4)(+) T cells was examined by real time quantitative polymerase chain reaction (PCR). RESULTS: After vaccination, the intraocular inflammation was significantly mitigated, antigen specific DTH and lymphocyte proliferation responses were suppressed, interleukin (IL)-2 in aqueous humour, interferon (IFN)-gamma and IL-2 produced by the spleen and draining lymph node cells were significantly decreased, whereas the production of IL-4 and IL-10 were increased. The response of draining lymph node cells to TCR V(beta) 8.3 peptide was enhanced after vaccination. Inoculation with CFA alone did not affect the severity of EAU and the above parameters. The suppression of EAU was much stronger in the group of four fold inoculations than the group of two fold inoculations. The expression of TCR V(beta) 8.3 gene was significantly reduced in the group of fourfold inoculations. CONCLUSION: Vaccination with the synthetic TCR V(beta) 8.3 peptide could remarkably inhibit the development of EAU. PMID- 16701015 TI - Follow-up of combined intervention for patients with both renal and cerebral artery stenosis. PMID- 16701016 TI - Expression of HLA class I and II on peripheral blood lymphocytes in HBV infection. PMID- 16701017 TI - Study of cultured bovine capsular bag in pure ocular tissue. PMID- 16701018 TI - Long-term results of elective open repair for abdominal aortic aneurysm. PMID- 16701019 TI - Short- and long-term outcomes of lung volume reduction surgery. PMID- 16701020 TI - Risk estimation based on mixed normal distribution model for diabetes-related hospitalization claims. PMID- 16701021 TI - Transcatheter closure of a giant coronary artery fistula with patent duct occluder. PMID- 16701022 TI - Primary pigmented nodular adrenocortical disease: report of 5 cases. PMID- 16701023 TI - Visceral and renal arteries stenosis associated with Takayasu arteritis. PMID- 16701024 TI - Deep brain stimulation in the treatment of tardive dystonia. PMID- 16701026 TI - [Further improvement of ergonomic applied studies in China]. PMID- 16701025 TI - [The course management and medical service in construction population at high altitude]. PMID- 16701027 TI - [Maximum acceptable weight of lift for manual lifting tasks]. AB - OBJECTIVE: To explore the maximum acceptable weight of lift (MAWL) in different lifting conditions and the suitable lifting equation for the stipulation of occupational health standard of manual lifting tasks in China. METHODS: The MAWL was investigated among the thirteen male and ten female students using psychophysical methodology and the recommended weight of limit (RWL) was compared. RESULTS: The MAWL of male and female subjects was decreased gradually with the increase of lifting height. Once the height of lifting was over shoulder, the MAWL was decreased dramatically. The RWL was greater than the MAWL at 25 cm horizontal distance in male subjects, but the RWL was smaller than the MAWL at the distance of 45 cm and 63 cm. The average MAWL of male subjects was 30.8% greater than that of female subjects. The MAWL was decreased gradually with increase of the horizontal distance at the same height of lifting. The result of the male subjects was consistent with that of the female subjects. For the asymmetric lifting tasks of the male subjects, the bigger the angle of rotation was, the less the MAWL became. The angle of rotation had negative correlation with the capability of lifting (r = -0.996 6, P < 0.01). When the subjects performed asymmetric lifting tasks, the RWL was smaller than the MAWL and the difference was significant. CONCLUSION: The revision is sufficient for the horizontal distance and asymmetric lifting in the NIOSH lifting equation, but it is not suitable for height, especially for the task of lifting over shoulder. The RWL for the task of lifting over shoulder should be lowered. The factor of gender should be taken into account in the lifting equation and the constant of gender S can be added. Then S = 1 for male while S = 0.692 for female. PMID- 16701028 TI - [Evaluation of work load and related factors during asymmetric lifting with surface electromyography]. AB - OBJECTIVE: To evaluate the work load and related factors for asymmetric lifting tasks using surface electromyography (EMG). METHODS: Thirteen male volunteers lifted loads of 6 and 13 kg at two speeds, in varying angles of trunk rotation, from floor to knuckle height, performing 16 lifting tasks. During lifting, surface electromyography signals from the erector spinae, bilaterally at level of T10 and L3, was continually recorded. The work load and related factors for asymmetric lifting tasks were evaluated by comparing the average amplitude of EMG signals. RESULTS: The EMG average amplitude when lifting the load of 13 kg (43.30% MVE) was significantly greater than that when lifting the load of 6 kg (37.70% MVE) (P < 0.01). The EMG average amplitude when lifting at the higher speed (41.80% MVE) was significantly greater than that at the lower speed (39.19% MVE) (P < 0.05). The EMG average amplitude was increased with the increase of the trunk rotation angle without significant difference (P > 0.05). At T10, the EMG average amplitude was greater on the right side than that on the left side (50.31% MVE and 25.88% MVE respectively) while at L3, it was on the contrary (45.60% MVE on the left and 40.22% MVE on the right respectively) (P < 0.01). Multiple stepwise regression analysis showed that the load of lifting was the factor contributing most to the EMG average amplitude followed by the lifting speed. CONCLUSION: The risk factors related to work load for asymmetric lifting tasks are mainly the load and the speed of lifting. The EMG activity between the right and left side of erector spinae is different during such lifting. PMID- 16701029 TI - [Characteristics of surface electromyography and work load of the forearm extensors in repetitive wrist extending]. AB - OBJECTIVE: To evaluate the influence of the frequency, the weight and the motion angle on the stress and the fatigue of the forearm extensors in repetitive wrist extending at low force loading level with surface electromyography (SEMG). METHODS: Sixteen male college student volunteers were recruited for the experiment. Eight tasks of wrist extending were performed for 20 minutes respectively in given weight (1.96, 4.90 N), frequency (8.0, 33.3 moves/minute) and motion angle (45 degrees, 90 degrees). The static wrist extending at the level of 20% maximum voluntary contraction (MVC) were performed before and after each task for 2 up to 3 seconds, and the SEMG signals of extensor carpi ulnaris muscle (ECU) and extensor digitorum (ED) were recorded and analyzed. RESULTS: The weight loading level was approximately equal to 1.40% or 3.50% of the MVC force. The mean power frequency (MPF) and the median frequency (MF) were decreased with the increase of 3 kinds of loading levels. The decrease of MPF of the muscle ED was significant (P < 0.05 or P < 0.01). The MF was decreased with the increase of angle and weight loading levels (P < 0.05 or P < 0.01). The root mean square (RMS) value of SEMG could be divided into 3 or 4 groups with significant difference. All three kinds of loads had positive correlation with amplitude of SEMG according to the stepwise regression analysis. CONCLUSION: The fatigue level of ED is the highest. The primary load factor for the forearm extensors is the frequency followed by the weight and the angle. MF, MPF and RMS can be used as sensitive indexes for evaluating the stress and the fatigue of the forearm extensors during repetitive performance at lower force loading level. PMID- 16701030 TI - [The role of JWA in N-methyl-N'-nitro-N-nitrosoguanidine induced human bronchial epithelial cell apoptosis]. AB - OBJECTIVE: To investigate the role and possible mechanism of JWA in N-methyl-N' nitro-N-nitrosoguanidine (MNNG) induced human bronchial epithelial (HBE) cell apoptosis. METHODS: The cell growth inhibition rate was detected by MTT, the cell apoptosis was measured by Hoechst staining, the expression of JWA protein was detected by Western blot, and the potential binding protein of JWA proximal promoter was detected by Southwestern assay. RESULTS: MNNG treatment of HBE cells for 24 hours induced apoptosis with significant dose-effect relationship and in this course the expression of JWA protein was elevated. The 2.0 microg/ml MNNG treated cells for 24 hours activated nuclear transcription factor expression that specifically bound to JWA proximal promoter. CONCLUSION: That MNNG treatment activates nuclear transcription factor binding to JWA proximal promoter may be involved in intracellular apoptosis associated signal pathway. PMID- 16701031 TI - [Effects of hemin and thermal stress exposure on JWA expression]. AB - OBJECTIVE: To investigate the expression of JWA after hemin and (or) thermal stress exposure. METHODS: The expression of JWA was determined by Western blot. RT-PCR was carried out for evaluation of the expression of JWA mRNA. The JWA promoter transcription activity analysis was performed by CAT-ELISA. RESULTS: The expression of JWA protein was significantly increased by 3.23 +/- 0.57 times of control in K562 cells after treated by hemin (50 micromol/L) for 1 week, and the similar pattern was observed in the cells after treatment with thermal stress (42 degrees C) for 2 hours (increased by 8.00 +/- 1.73 times). The expression of JWA mRNA was also significantly elevated by 1.37 +/- 0.06 times in K562 cells treated by hemin (30 micromol/L) for 48 hours, and with a similar regulation pattern (increased by 1.87 +/- 0.13 times) by treatment with thermal stress (42 degrees C) for 30 minutes. However, an antagonistic effect was observed by treatment of K562 cells with hemin (30 micromol/L, 48 hours) followed by thermal stress (42 degrees C, 30 minutes). The CAT-ELISA further confirmed that hemin or thermal stress treatment alone up-regulated JWA transcription activity while the effects could be counteracted partly by the combined treatment of the both. CONCLUSION: The hemin and thermal stress may regulate JWA expression via distinct intracellular signal transduction pathways. PMID- 16701032 TI - [Effect of dimethoate on the expression of heat shock protein 70 in peripheral blood lymphocytes of human beings]. AB - OBJECTIVE: To study the effect of dimethoate on the expression of heat shock protein 70 (HSP70) in peripheral blood lymphocytes of human beings and to explore the feasibility of HSP70 in biomonitoring among workers exposed to organophosphorous pesticides. METHODS: Peripheral blood lymphocytes were isolated from subjects, comprising 11 people of the control group and 35 workers of the exposure group exposed to dimethoate. Flow cytometry was used for detecting both the basic level and the level of the dimethoate-induced expression of HSP70. The activity of whole blood acetylcholinesterase (AChE) was examined at the same time. Then the potential influential factors to HSP70 expression and AChE activity were analyzed. RESULTS: The basic level of HSP70 expression in the exposure group and the control group was 41.24% +/- 10.45% and 23.97% +/- 4.29% respectively. The activity of AChE in these two groups were (125.23 +/- 7.97) and (145.36 +/- 8.78) U/ml respectively. Both differences were statistically significant (P < 0.01). Among the exposure group, the basic level of HSP70 expression of the two categories comprising operators and packers, were 47.34% +/ 11.87% and 38.05% +/- 8.20% respectively (P < 0.05), while there was no significant difference (P > 0.05) in AChE activity between these two categories. The factors that had significant influence on the HSP70 basic level of the exposure group were the health condition, the environmental concentration of dimethoate and the exposure time in order, according to their significance of influence. At least 88% variance of HSP70 could be explained by these factors. The only factor that could influence AChE activity significantly was the exposure time, and it could only explain about 12% variance of AChE activity. After the treatment of dimethoate in vitro, the level of the induced expression of HSP70 in the control group was significantly higher than that of the exposure group (P < 0.01). The increasing order was the control group, the group of packers and the group of operators according to the increasing extent and there were significant difference among them (P < 0.01). The factors that could significantly influence the change ratio of HSP70 expression were the environmental concentration and the exposure time. CONCLUSION: HSP70 is a potential index that can reflect the individual and environmental conditions of workers exposed to dimethoate comprehensively. PMID- 16701034 TI - [Effect of exercise stress on cigarette smoking induced downregulation of BKca and Kv1.5 expression in pulmonary arterial smooth muscle cells of rats]. AB - OBJECTIVE: To investigate the effect of exercise stress on chronic cigarette smoking induced downregulation of large conductance calcium-activated potassium channel (BKca) and voltage-dependent delayed rectifier potassium channel (Kv1.5) expression in pulmonary arterial smooth muscle cells of rats. METHODS: Rats were divided into three groups: the normal control group, the smoking control group and the smoking + exercise group. The plasma cortisol level, the potassium channel expression and the pathological changes in lung tissue were determined with HE staining, the immunohistochemistry and the in-situ hybridization. RESULTS: (1) In the smoking + exercise group, the plasma cortisol level was determined immediately after exercise [(1528.7 +/- 469.7) ng/L] and was higher than that determined before exercise [(672.4 +/- 235.7) ng/L] (P < 0.01); (2) The HE staining showed that the chronic pulmonary inflammatory response in the smoking control group was severe while it was mild in the smoking + exercise group; (3) The mRNA and protein expression (OD value) of BKca in the smoking control group (mRNA: 0.2206 +/- 0.0415 for big artery and 0.3935 +/- 0.1378 for small artery; protein: 0.2634 +/- 0.1219 for big artery and 0.0995 +/- 0.0851 for small artery) were less than those in the normal control group. The mRNA expression of BKca in the smoking + exercise group (OD value) (0.5022 +/- 0.1134 for big artery and 0.6408 +/- 0.2135 for small artery) was higher than that in the smoking control group; (4) The mRNA and protein expression of Kv1.5 in the smoking control group (OD value) (mRNA: 0.9354 +/- 0.3290 for big artery and 0.5012 +/- 0.1170 for small artery; protein: 1.1112 +/- 0.3310 for big artery and 0.4736 +/- 0.1250 for small artery) were less than those in the normal control group. The protein expression of Kv1.5 in the smoking + exercise group (0.7445 +/ 0.2690) in small artery was higher than that in the smoking control group. CONCLUSION: Proper exercise stress can decrease inhibition effect of the chronic smoking on the expression of potassium channel BKca and Kv1.5, which perhaps partly results from exercise induced increase of cortisol secretion. PMID- 16701035 TI - [Effect of 1.8 GHz radiofrequency electromagnetic fields on the expression of microtubule associated protein 2 in rat neurons]. AB - OBJECTIVE: To investigate the changes of gene expression in rat neurons induced by 1.8 GHz radiofrequency electromagnetic fields (RF EMF) and to screen for the RF EMF-responsive genes. METHODS: Newly-born SD rats in 24 hours were sacrificed to obtain cortex and hippocampus neurons. The cells were divided randomly into two groups: the experiment group (the irradiation group) and the control group (the false irradiation group). In the irradiation group, after twelve days' culture, neurons were exposed to 1.8 GHz RF EMF modulated by 217 Hz at a specific absorption rate (SAR) of 2 W/kg for 24 hours (5 minutes on/10 minutes off) while in the false control group, the neurons were put in the same waveguide as in the irradiation group, but were not exposed to any irradiation. The total RNA was isolated and purified immediately after exposure. The affymetrix rat neurobiology U34 assay was used for detecting the changes in gene expression profile according to the manufacturer's instruction. RF EMF-responsive candidate gene was confirmed by using ribonuclease protection assay (RPA). RESULTS: Among 1200 candidate genes, the expression levels of 34 genes were up or down regulated. Microtubule associated protein 2 (Map2) gene was selected as the candidate and subjected to further analysis. RPA data clearly revealed that Map2 was statistically significantly up-regulated after neurons were exposed to the RF EMF (P < 0.05). CONCLUSION: The modulation of gene expression and function of Map2 as a neuron specific cytoskeleton protein is crucial to maintain the normal framework and function of neurons. The finding that 1.8 GHz RF EMF exposure increases the expression of Map2 might indicate some unknown effects of RF EMF on neurons. PMID- 16701037 TI - [Reliability and validity of Burnout Questionnaire in occupational population]. AB - OBJECTIVE: To evaluate the reliability and validity of simple Burnout Questionnaire in occupational population. METHODS: A self-administered Burnout Questionnaire comprising 19 items was developed in light of Maslash Burnout Questionnaire and used for investigating 458 employees. RESULTS: Cronbach's alpha between total items of Burnout Questionnaire and each factor fell in between 0.82 and 0.85 through the consistency test. All 19 items of Burnout Questionnaire were subjected to factor analysis, and three latent factors were identified, wherein 56.3% of total variance could be explained. According to the contents described in the items and Maslash burnout theory, they were emotional exhaustion, depersonalization, and personal accomplishment. According to the covariance and the variance analysis, there was significant difference in the integral of the type of work among the three subitems (P < 0.01); there was significant difference in the integral of depersonalization between two sexes (P < 0.01); There were significant difference in the integral of the personal achievement among different level of education (P < 0.01). CONCLUSION: The reliability and validity of Burnout Questionnaire is acceptable and can be used for assessing burnout in occupational population. PMID- 16701036 TI - [Effects of artesunate on CD14 and toll-like receptor 4 in peritoneal macrophages of mice with heat stroke endotoxemia]. AB - OBJECTIVE: To study the effects of artesunate on CD14 and toll-like receptor 4 (TLR 4) expressions in peritoneal macrophages of mice with heat stroke endotoxemia. METHODS: Kunming mice were randomly divided into the normal temperature group, the hyperthermia group, the normal saline (NS) group and the artesunate group (both i.p.60 mg/kg daily for consecutive five days). The normal temperature group was exposed to the condition of dry bulb temperature (Tdb) 25 degrees C +/- 0.5 degrees C and relative humidity (RH) 43% +/- 5% for 2 hours, while other groups were exposed to the condition of Tdb 35 degrees C +/- 0.5 degrees C and RH 65% +/- 5%. The mRNA expressions of CD14 and TLR 4 in peritoneal macrophages and concentrations of tumor necrosis factor alpha (TNF alpha) in plasma were observed in different time points (1 hour and 2 hour). RESULTS: The mRNA expressions of CD14 and TLR 4 in the normal temperature group were 0.34% +/- 0.047% and 0.31% +/- 0.062% respectively. The expressions of two receptors at 1 hour in the hyperthermia group were significantly increased to 0.53% +/- 0.085% and 0.45% +/- 0.049% compared with the normal group and kept increased at 2 hour (P < 0.01). The mRNA expressions at 1 hour in the NS group were significantly increased but a little bit decreased at 2 hour. The mRNA expressions of CD14 and TLR 4 at 1 hour in the artesunate group were 0.26% +/- 0.051% and 0.25% +/- 0.084% respectively and a little bit decreased at 2 hour. The change of TNF-alpha in each group was almost consistent with the changes of CD14 and TLR 4. CONCLUSION: Artesunate can reduce significantly the expressions of CD14 and TLR 4 in LPS signal transduction pathway and the concentration of TNF-alpha, which perhaps is one of the most important mechanisms that artesunate fights against endotoxemia. PMID- 16701038 TI - [Protective effect of heat preconditioning on NIH-3T3 fibroblast]. AB - OBJECTIVE: To establish stress adaptation model of mouse fibroblast cell line NIH 3T3, to provide a group of parallel object for stress adaptation research, and to explore the function and mechanism of HSP90 in stress adaptation. METHODS: A stress-adapted cell model was established by thermal preconditioning (42 degrees C, 20 minutes), and the adaptation result was evaluated by observing the change of the membrane injury and the damage of DNA induced by the heat stress for the second time (44 degrees C, 20 minutes). The HSP90 content was detected by Western blot. RESULTS: According to the membrane injury and HSP90 synthesis induced by the heat stress for the second time, it was primarily confirmed that 6 hours after thermal preconditioning were the optimum stress protection time. When cells underwent heat stress for the second time 6 hours after thermal preconditioning, the membrane injury (15.4% +/- 2.6% vs 41.2% +/- 5.1%), damage of DNA (15.1% vs 26.3%) were decreased compared with the control group in which there was no preconditioning. The OD(HSP90)/OD(control) value indicated that the cellular HSP90 contents was decreased immediately after heat stress (44 degrees C, 40 min). The content of HSP90 was 0.82 +/- 0.18 in the heat stress group, 1.70 +/- 0.52 in the preconditioning group and 1.41 +/- 0.16 in the heat stress after preconditioning group. CONCLUSION: With the preconditioning for the NIH-3T3, the time point for the stress protection is confirmed, the model for the cellular stress adaptation is established and the protective effect of HSP90 is primarily confirmed in this model. PMID- 16701039 TI - [Magnetic resonance imaging for the wrist joint of the coal miners in vibration department]. AB - OBJECTIVE: To study the magnetic resonance imaging (MRI) in the wrist joint of coal miners who work in excavation and vibration department. METHODS: Forty-three coal miners with the hand-arm vibration disease served as the observation group while 20 workers who were not working in the vibration department acted as the control group. The patients in the observation group were divided into five subgroups according to the time when they received vibration. The regularity of the development of signs and symptoms of MRI was observed and analyzed. RESULTS: The hydroarthrosis was most found in MRI. There were significant difference in hydroarthrosis (chi(2) = 8.80, P < 0.01), osteoporosis and osteomyelitis (chi(2) = 3.91, chi(2) = 5.01, P < 0.05 respectively) between the observation group and the control group. The edema of bone marrow and the avascular necrosis of ossa carpi were found only in the observation group and not found in the control group. The hydroarthrosis and the edema of bone marrow occurred most in the early stage of vibration. The signal in the edema of the bone marrow of the distal end of the radius was decreased in the GE sequence T(2)WI with the specificity. CONCLUSION: (1) Changes in the wrist joint occur in the early stage of the vibration work, and can be found in the MRI. (2) The edema of the bone marrow of the distal end of the radius is of great value in the diagnosis of the hand-arm vibration disease. PMID- 16701040 TI - [One patient with rupture of esophagus induced by paraquat poisoning]. PMID- 16701041 TI - [Effect of 1, 2-dichloroethane on blood brain barrier]. AB - OBJECTIVE: To investigate the effect of 1, 2-dichloroethane (1, 2-DCE) on blood brain barrier. METHODS: Acute toxic encephalopathy model was copied with the consecutive static inhalation of 1, 2-DCE. The water content of brain tissue was measured, and the blood brain barrier permeability was detected with lanthanum nitrate. The brain microvascular endothelial cells and neuroglial cells were cultured in vitro, which were administrated with 1, 2-DCE. The cell morphologic structures were observed under light microscope and electron microscope. RESULTS: (1) The extracellular edema was most found in the cerebral tissue and the leakage of lanthanum particles through the barrier were found with the lanthanum tracking method. (2) The water content in cerebral cortex in the moderate and high dose groups was significantly higher than that in the control group and became severer with the increases of the intoxicated time. The water content in cerebral medulla was significantly increased only at 6 hours after the intoxication. (3) The normal morphological structure of brain microvascular endothelial cells and neuroglial cells could be injured by 1, 2-DCE, and the injury to neuroglial cells caused by 1, 2-DCE occurred earlier and severer than that to brain microvascular endothelial cells. CONCLUSION: 1, 2-DCE can damage blood brain barrier and induce cerebral edema. PMID- 16701042 TI - [Survey of the 50 Hz electric field intensity distribution in drivers' cabs of the electric locomotives]. PMID- 16701043 TI - [Effect of high power microwave radiation on ultrastructure of neuron synapse and content of amino acid neurotransmitters in hippocampus of rats]. PMID- 16701044 TI - [Effects of psychosocial factors on work-related musculoskeletal disorders]. PMID- 16701045 TI - [Advance of research on skin injury after exposure to 2, 3, 7, 8 tetrachlorodibenzo-p-dioxin]. PMID- 16701047 TI - [Three patients with acute phosgene poisoning]. PMID- 16701046 TI - [Clinical analysis of seven patients with acute aluminum phosphide poisoning]. PMID- 16701048 TI - [Investigation on work-related musculoskeletal injury in pipe welding workers]. PMID- 16701049 TI - [An accident of acute dimethyl sulfate poisoning]. PMID- 16701050 TI - Catastrophic disaster. PMID- 16701051 TI - Treatment of open intra-articular calcaneal fractures: evaluation of a treatment protocol based on wound location and size. AB - BACKGROUND: Effective treatment algorithms for open, displaced, intra-articular calcaneal fractures and their potential early complications have not yet been established. This is a retrospective review of the management of open displaced calcaneal fractures at a Level 1 trauma center and their resulting soft-tissue complications in an effort to establish guidelines for management. METHOD: Between November, 1994, and April, 2002, 31 open intra-articular calcaneal fractures in 29 patients were treated at our institution. All fractures were treated with a standard protocol based on the appearance of the traumatic wound after initial debridement and stabilization. Fracture stabilization was determined by the wound characteristics and was either standard internal fixation or percutaneous fixation. Data on associated injuries, comorbidities, smoking, and subsequent soft-tissue complications were collected and reviewed. RESULTS: Overall there was a 29% soft-tissue complication rate with this protocol. Two of the four laterally based traumatic wounds had complications. Twenty-seven wounds were medial. Nine were considered unstable and were treated with aggressive wound management and percutaneous fixation of the tuberosity reduction; two developed deep infections and required amputation and seven were salvaged with patients returning to full weightbearing. Eighteen fractures with medial wounds were treated with standard internal fixation; soft-tissue complications occurred in five. Initial size greater than 4 cm was associated with an increased complication rate in this group. CONCLUSION: Management of open calcaneal fractures and the risk of complications depend on the size and position of the traumatic wound. Lateral wounds are rare and in this review, two of the four had complications using this protocol. Medial wounds of less than 4 cm can be treated with open reduction and standard internal fixation if the wound can be closed and remain stable off antibiotics. Larger wounds (more than 4 cm) or unstable wounds should not be treated with open reduction and internal fixation but can be reduced and held in alignment with percutaneous wire fixation. PMID- 16701052 TI - Treatment of Eichenholtz stage I Charcot foot arthropathy with a weightbearing total contact cast. AB - BACKGROUND: Initial treatment of Eichenholtz stage I Charcot arthropathy of the foot generally is total contact casting and nonweightbearing. This method, however, often is time-consuming and has a poor result. This study was done to determine the success rate of total contact casting in a small group of patients and to establish a benchmark time period for treatment. METHODS: Ten subjects with acute Eichenholtz stage I (stage of development) Charcot foot arthropathy were prospectively treated with weightbearing total contact cast therapy, undergoing biweekly cast changes. One subject did not complete the study. Subjects were monitored with biweekly clinical examination, limb volume measurement, and radiographs. The average age was 58.2 (range 39 to 72) years and weight was 216.9 (range 160 to 275) pounds. RESULTS: All subjects were able to use commercially available depth-inlay shoes and custom accommodative foot orthoses at an average of 9.2 (range 8 to 16) weeks. One subject developed a superficial ulcer that resolved with footwear modification. CONCLUSION: This preliminary study supports the use of total contact cast therapy and weightbearing in the treatment of acute Charcot foot arthropathy. When the total contact cast was changed every 14 days, all subjects were able to use commercially available depth-inlay shoes and custom orthoses. PMID- 16701053 TI - Rupture of the anterior tibial tendon: three clinical cases, anatomical study, and literature review. AB - BACKGROUND: We report three cases of anterior tibial tendon ruptures and the results of an anatomical study in regard to the tendon's insertion site and a literature review. METHODS: Three patients were referred to our hospital with anterior tibial tendon ruptures. In the anatomical study, 53 feet were dissected, looking in particular for variants of the bony insertion of the tendon. RESULTS: Two patients had surgical treatment (one primary repair and one semimembranosus tendon graft) and one conservative treatment. After a mean followup of 14 weeks all patients had satisfactory outcomes. In the anatomical study, we noted three different insertion sites: in 36 feet the tendon inserted into the medial side of the cuneiform and the base of the first metatarsal bone and in 13 feet only into the medial side of the cuneiform bone. In the remaining four feet the tendon inserted into the cuneiform and the first metatarsal bone, but an additional tendon was noted taking its origin from the anterior tibial tendon near its insertion into the medial cuneiform and attaching to the proximal part of the first metatarsal. CONCLUSIONS: According to literature, surgical repair is the treatment of choice for acute ruptures and for patients with high activity levels. For chronic ruptures and patients with low demands, conservative management may lead to an equally good outcome. Knowledge of the anatomy in this region may be helpful for diagnosis and for the interpretation of intraoperative findings and choosing the most appropriate surgical procedure. PMID- 16701054 TI - Comparison of plate and screw fixation and screw fixation alone in a comminuted talar neck fracture model. AB - BACKGROUND: Talar neck fracture fixation has been studied in noncomminuted fracture models, but no large clinical series of comminuted fracture patterns have been published and no biomechanical studies have compared plate fixation with screw fixation in comminuted talar neck fractures. METHODS: Nine matched pairs of fresh frozen talar specimens were stripped of soft tissue and mounted in a cylindrical jig. The talar neck was fractured using a dorsally directed shear force at a rate of 200 mm/min, and dorsal comminution was simulated by removing a 2-mm section of bone from the distal fracture fragment. One specimen from each pair was fixed with either two solid 4.0-mm partially threaded cancellous screws posterior-to-anterior just lateral to the posterior process of the talus or with a four-hole 2.0-mm minifragment plate contoured to the lateral surface of the talar neck and secured with 2.7-mm screws. A 2.7-mm fully threaded cortical screw was placed medially using a lag technique. The specimens were then loaded to failure with a dorsally directed force at a rate of 200 mm/min. Failure was defined as the load producing 2 mm of displacement. A Student's t-test analysis was used with significance set at p < or = 0.05. RESULTS: Posterior-to-anterior screw fixation had a statistically significant higher load to failure than plate fixation (p < 0.05). Mean load to failure for the screw group was 120.7 +/- 68.5 N and 89.7 +/- 46.6 N for the plating group. CONCLUSIONS: Plate fixation may offer substantial advantages in the ability to control the anatomic alignment of comminuted talar neck fractures, but it does not provide any biomechanical advantage compared with axial screw fixation. Further, the fixation strength of both methods was an order of magnitude lower than those found in previous studies of noncomminuted fractures. PMID- 16701055 TI - Characterization of lesions of the talus and description of tram-track lesions. AB - BACKGROUND: The ankle joint, although not prone to primary osteoarthritis (OA), is known to be susceptible to secondary OA as a result of sports injuries and other trauma. Unlike the knee joint, a thorough investigation of talar cartilage lesions has not been previously reported. METHODS: One hundred and five human tali from 67 donors were used to determine the type and location of the most common lesions through gross examination, radiography, diffraction enhanced imaging, and histology. "tram-track lesions" also are described. RESULTS: The most anterior and posterior regions of the talar dome, along with the medial and lateral borders of the dome were most affected by cartilage degeneration. These are regions that appear to be most subjected to frictional forces from their articulating counterparts during high stress activities that move articulating surfaces slightly out of congruence with each other. One particularly striking cartilage degeneration pattern that, to our knowledge, has not been described histologically, is the tram-track lesion. These lesions displayed a longitudinally oriented groove in the cartilage, running from anterior to posterior, in which the deepest portion of the groove was located in the middle of the anterior-posterior axis of the talus. Several of these cartilage grooves had a bony ridge beneath, mirroring the cartilage groove whereas others did not. This suggests that the cartilage groove develops before the interruption of the tidemark (border between the calcified and uncalcified cartilage). In specimens for which the articulating tibial articular surface was available, (17) it was found that the tibia displayed small osteophytes on the anterior articular margin that exactly corresponded to their articulation with the talar cartilage grooves as the ankle articulated through plantar flexion and dorsiflexion. CONCLUSION: This study provides an in-depth histologic and gross anatomic look at the most common lesions of the talus of the ankle joint. In particular, the "tram-track" lesion was shown to be a consequence of its tibial articulation and to include both subchondral bone and articular cartilage changes. PMID- 16701056 TI - Classification of postaxial polydactyly of the foot. AB - BACKGROUND: The origins and shapes of accessory digits in postaxial polydactyly of the foot were analyzed morphologically and radiographically, and their characteristics were determined. A simple classification method was then devised to assist in determining the most appropriate treatment options. METHODS: We evaluated 113 feet of 95 patients who had surgery for the treatment of postaxial polydactyly between 1998 and 2002. Based on the morphologic, radiographic, and operative findings, the cases were classified according to the origin of the accessory digit: middle phalangeal, proximal phalangeal, floating, fifth metatarsal, or fourth metatarsal. The proximal phalangeal type was further divided into three subtypes: proximal phalangeal lateral type, proximal phalangeal medial, and proximal phalangeal head. RESULTS: Of the 113 feet, 36 were middle phalangeal type, 45 were proximal phalangeal type, 5 were floating type, 15 were fifth metatarsal type, and 12 were fourth metatarsal type. Of the proximal phalangeal types, 15 were laterally duplicated supernumerary sixth digits, and 17 were medially duplicated supernumerary fifth digits. The duplicated digits of the remaining 13 originated at the distal portion of the proximal phalanx. In the middle phalangeal, proximal phalangeal head, proximal phalangeal medial, and fourth metatarsal types, the medial accessory fifth digit was an abnormally duplicated digit, which was excised. In the proximal phalangeal lateral, floating, and fifth metatarsal types, the lat eral accessory sixth digit was excised. For the children in this study, we did not perform reconstruction of the deep transverse metatarsal ligament or collateral ligament. Also, we did not use longitudinal pin fixation. Skin necrosis occurred in 10 feet that resolved, and in five of the 15 feet of the 5th metatarsal medial deviation occurred. CONCLUSIONS: Based on the morphologic, radiographic, and operative findings, we suggest a classification method of postaxial polydactyly of the foot. We believe this is a straightforward and useful method for the treatment of postaxial polydactyly. PMID- 16701057 TI - An in vitro study comparing the use of suture anchors and drill hole fixation for flexor digitorum longus transfer to the navicular. AB - BACKGROUND: The surgical management of posterior tibial tendon dysfunction often includes transfer of the flexor digitorum longus (FDL) tendon through a tunnel in the navicular. Fixation often is obtained by sewing the tendon back onto itself. The purpose of this study was to compare this standard method of fixation with suture anchor fixation, a technique that may be associated with less surgical morbidity, because it requires the harvesting of less tendon length. METHODS: FDL tendon transfer to the navicular was done in 13 fresh-frozen cadaver specimens. In six feet comprising the standard group, the FDL tendon was transected distal to the master knot of Henry, placed through a drill hole into the navicular, and sutured back onto itself. In seven feet the FDL tendon was transected proximal to the master knot of Henry, placed into a drill hole into the navicular, and fixed with a suture anchor. Load was applied to the proximal FDL muscle and tendon using a materials testing system (MTS) machine and peak load to failure was measured. RESULTS: The mean load to failure was 142.48 N +/- 38.06 N for the standard group and 142.12 N +/- 59.26 N for the suture anchor group (p = 0.305 for the Student-t test and p = 0.945 for the Mann-Whitney test). CONCLUSION: Transfer of the FDL tendon to the navicular using suture anchor fixation requires less tendon length yet provides similar fixation strength as compared to sewing the tendon back onto itself. However, suture anchors are considerably more expensive than sutures. CLINICAL IMPLICATIONS: Suture anchors allow comparable fixation of FDL tendon transfer into a navicular without the need to disrupt the master knot of Henry. This technique may be associated with less morbidity including a shorter incision, decreased risk of medial plantar nerve injury, and decreased loss of lesser toe plantarflexion strength secondary to maintenance of the normal interconnections between the flexor hallucis longus (FHL) and FDL tendons. PMID- 16701058 TI - The effect of gender, age, and lateral dominance on arch height and arch stiffness. AB - BACKGROUND: Arch structure is known to vary widely. However, it may be linked to intrinsic factors such as gender, age, and lateral dominance. Understanding the association between these factors and arch structure may be useful in understanding injury biases that exist between individuals with different foot types. METHODS: The foot structure of 145 subjects, 68 men and 77 women (18 to 65 years) was examined in this study. The arch height index, a measure of dorsal height normalized to foot length, and arch stiffness of both feet were measured in each subject. Comparisons of both arch height and arch stiffness were made between genders and between the dominant and nondominant feet. In addition, the relationship between both arch height and stiffness and age was examined. RESULTS: There was no difference between the arch height index of men and women; however, the arches in women were significantly less stiff (p = 0.00). There were no statistically significant relationships between increasing age and either arch height index or stiffness. The within-subject comparisons showed that the dominant foot had a significantly higher arch height index than the nondominant foot (p = 0.00). However, arch stiffness was not different between sides. There was a significant, but weak, relationship between arch height index and arch stiffness (p = 0.00, R2 = 0.09) with a higher arch height index corresponding to a stiffer arch. CONCLUSION: Understanding differences in arch structure may lend insight into the predilection for injury between genders, with increasing age, and between sides of a given subject. PMID- 16701059 TI - Stress fracture in the presence of a calcaneonavicular coalition: a case report. PMID- 16701060 TI - Malignant fibrous histiocytoma associated with remote internal fixation of an ankle fracture. PMID- 16701061 TI - Mini L-shaped incision for calcaneal osteotomy: clinical tip. PMID- 16701062 TI - Ray amputation with limited incision: clinical tip. PMID- 16701063 TI - Current concepts review: foot and ankle outcome instruments. PMID- 16701064 TI - Special report: highlights of the Twenty-first Annual Summer Meeting of the American Orthopaedic Foot and Ankle Society, Boston, Massachusetts, July 15-17, 2005. PMID- 16701065 TI - Assessment of diagnostic tools for eradication of bovine tuberculosis in cattle co-infected with Mycobacterium bovis and M. avium subsp. paratuberculosis. AB - The intradermal tuberculin (IDTB) test and the interferon-gamma (IFN-gamma) assay are used worldwide for detection of bovine tuberculosis in cattle, but little is known about the effect of co-infecting agents on the performance of these diagnostic tests. This report describes a field trial conducted in a cattle herd with dual infection (bovine tuberculosis and paratuberculosis) during 3.5 years. It has been based on a strategic approach encompassing serial parallel testing (comparative IDTB test, the IFN-gamma assay and serology of paratuberculosis) that was repeated 8 times over the period, and segregation of animals into two herds. The IDTB test detected 65.2% and the IFN-gamma test detected 69.6% of the Mycobacterium bovis culture-positive cattle. However, the IDTB test performed better during the first part of the trial, while the IFN-gamma test was the only method that detected infected animals during the following three samplings. The number of false positive reactors with the IDTB and/or the IFN-gamma tests was remarkably high compared to other reports, and could be caused by cross reactivity with M. avium subsp. paratuberculosis. Also, the M. bovis isolates from cattle and wildlife from the same property were characterised using molecular techniques to disclose an epidemiological link. The IDTB test may not be appropriate to eradicate bovine tuberculosis in herds with dual mycobacterial infections. This report highlights the need to use several diagnostic techniques for the accurate detection of M. bovis infected animals in these herds. PMID- 16701066 TI - Haemonchus contortus (Nematoda: Trichostrongylidae) infection in lambs elicits an unequivocal Th2 immune response. AB - Selection of resistant animals and host immunization have been proposed as alternative methods for the control of gastrointestinal nematode parasites. However, a better knowledge of the mechanisms involved in protective immunity against these parasites is required for the development of optimal strategies. In this study, 3 month old INRA 401 lambs (n = 81) were allocated into three groups: uninfected control, challenged either once (primary-infected animals) or twice (previously-infected animals) with 10,000 Haemonchus contortus L3. Uninfected control and challenged animals were sequentially sacrificed at 3, 7, 15 and 28 days post challenge. In both challenged groups, a clear Th2-oriented immune response was observed in the abomasal lymph node and mucosa. IL-4 and IL-13 mRNA over-expression, recruitment of eosinophils, mast cells and globule leukocytes and production of specific systemic IgG and mucosal IgA were observed earlier in previously-infected animals than in primary-infected ones. At 28 days post infection, no differences between intensities of these responses were observed between the challenged groups. Worm establishment rates were similar in previously-infected and primary-infected lambs. However, reductions of worm development, female fecundity and fecal egg output were observed in previously infected sheep. We conclude that H. contortus infection in young INRA 401 lambs induced an unequivocal Th2 immune response resulting in the regulation of worm egg production without affecting their establishment. PMID- 16701067 TI - Evaluation of a recombinant enzyme-linked immunosorbent assay for detecting Chlamydophila psittaci antibodies in turkey sera. AB - Chlamydophila psittaci (formerly Chlamydia psittaci) is one of the major pathogens associated with turkey respiratory disease. Devastating outbreaks with high mortality rates, similar to those of 1950 to 1970 in the USA occasionally occur, but respiratory signs without or with low mortality mostly characterize outbreaks now a day. Accurate diagnostic methods should be made available. The present study examined the sensitivity and specificity of a recombinant ELISA (rMOMP ELISA) for detecting Cp. psittaci major outer membrane specific antibodies in turkey sera. Test results were compared to those of immunoblotting and of a competitive ELISA (Chlamydia-psittaci-AK-EIA, Rohm Pharma, Germany) and an indirect ELISA (LPS/LGP) detecting antibodies to the lipopolysaccharide/lipoglycoprotein complex. The rMOMP ELISA was most sensitive as determined on serial dilutions of positive control sera originating from experimentally infected SPF turkeys. The competitive ELISA gave false positives since three negative controls reacted positive. For conventional sera, the sensitivities of the competitive ELISA, immunoblotting and the indirect ELISA were found to be 99.4, 93.1 and 82.2%, respectively, as compared to the rMOMP ELISA (100%). The specificities of the rMOMP ELISA, immunoblotting and the indirect ELISA were found to be 100% while the specificity of the competitive ELISA was only 2.7%. The rMOMP ELISA was chosen to compare the prevalence of chlamydiosis in 2002 with the one from 1992. In 2002, 188 on 200 (94%) turkey sera reacted positive compared to 175 on 200 (87.5%) in 1992 and like 10 years ago all examined farms were seropositive at slaughter. Interestingly, Belgian as well as French farms were seropositive. PMID- 16701068 TI - Chemiluminescent-based method for heme determination by reconstitution with horseradish peroxidase apo-enzyme. PMID- 16701069 TI - Two-dimensional electrophoretic mobility shift assay: identification and mapping of transcription factor CTCF target sequences within an FXYD5-COX7A1 region of human chromosome 19. AB - An approach for fast identification and mapping of transcription factor binding sites within long genomic sequences is proposed. Using this approach, 10 CCCTC binding factor (CTCF) binding sites were identified within a 1-Mb FXYD5-COX7A1 human chromosome 19 region. In vivo binding of CTCF to these sites was verified by chromatin immunoprecipitation assay. CTCF binding sites were mapped within gene introns and intergenic regions, and some of them contained Alu-like repeated elements. PMID- 16701070 TI - Whole cell immobilized amperometric biosensor based on Saccharomyces cerevisiae for selective determination of vitamin B1 (thiamine). AB - A new amperometric whole cell biosensor based on Saccharomyces cerevisiae immobilized in gelatin was developed for selective determination of vitamin B1 (thiamine). The biosensor was constructed by using gelatin and crosslinking agent glutaraldehyde to immobilize S. cerevisiae cells on the Teflon membrane of dissolved oxygen (DO) probe used as the basic electrode system combined with a digital oxygen meter. The cells were induced by vitamin B1 in the culture medium, and the cells used it as a carbon source in the absence of glucose. So, when the vitamin B1 solution is injected into the whole cell biosensor system, an increase in respiration activity of the cells results from the metabolic activity and causes a decrease in the DO concentration of interval surface of DO probe related to vitamin B1 concentration. The response time of the biosensor is 3 min, and the optimal working conditions of the biosensor were carried out as pH 7.0, 50mM Tris HCl, and 30 degrees C. A linear relationship was obtained between the DO concentration decrease and vitamin B1 concentration between 5.0 x 10(-3) and 10( 1) microM. In the application studies of the biosensor, sensitive determination of vitamin B1 in the vitamin tablets was investigated. PMID- 16701071 TI - Identification of a new chromophoric substrate in the library of amino acid p nitroanilides for continuous assay of VanX, a D,D-dipeptidase essential for vancomycin resistance. AB - As one of key bacterial proteins involved in vancomycin resistance, VanX is a D,D dipeptidase that impedes bacterial cell wall biosynthesis by hydrolyzing the essential D-Ala-D-Ala dipeptide. Based on a report by Crowder and co-workers that L-alanine-p-nitroanilide (L-Ala-pNA) was a useful substrate for continuous assay of VanX, we constructed a library of 35 L- and D-amino acid p-nitroanilides to provide the needed diversity to discover new substrates that are more specific than L-Ala-pNA. We report here that, among all compounds tested, D-leucine-p nitroanilide (D-Leu-pNA) was found to be the best substrate for VanX enzyme (KM=8.9+/-1.2 mM, kcat=0.0102+/-0.0016 s(-1), kcat/KM=0.0012 mM(-1)s(-1)). Although it is catalytically inefficient, this new VanX substrate needs essentially no sophisticated synthetic chemistry for preparation and therefore offers a convenient means for routine analysis of enzyme catalysis and the screening of potential inhibitors. Moreover, because it is the uncommon leucine in its D form in D-Leu-pNA, enzymatic activities due to other contaminated species in Escherichia coli used for VanX overproduction should be greatly reduced. PMID- 16701072 TI - Lowry method for the determination of pegylated proteins: the error, its reason, and a method for eliminating it. PMID- 16701073 TI - Phi29-based amplification of small genomes. PMID- 16701074 TI - Recovery of free oligosaccharides from derivatives labeled by reductive amination. AB - This study examined chemical regeneration of free oligosaccharides from their fluorescent derivatives prepared by reductive amination with various aromatic amines. Maltose derivatives of ethyl 4-aminobenzoate (p-ABEE), 2 aminobenzonitrile (o-ABN), 4-aminobenzonitrile (p-ABN), 7-amino-4-methylcoumarin (AMC), 2-aminobenzoic acid (o-ABA), 2-aminobenzamide (o-ABAD), 2-aminopyridine (AP), and 8-aminonaphthalene-1,3,6-trisulfonate (ANTS) were incubated at 30 degrees C with an aqueous solution of hydrogen peroxide/acetic acid. Recoveries of maltose from p-ABEE, p-ABN, and AMC derivatives were fairly good and gave approximately 90% of maltose. Recoveries of maltose from its o-substituted aniline (o-ABA, o-ABAD, and o-ABN) derivatives were 5-40%, but maltose was unrecoverable from AP and ANTS derivatives. Nevertheless, prior treatment of an AP derivative with cyanogen bromide enabled the regeneration of maltose in high yields. As an application, p-ABEE-labeled N-glycans from some glycoproteins separated on an amide column were identified by converting peak components to their AP derivatives via free saccharides and following mapping by reversed-phase chromatography. PMID- 16701075 TI - Masking oligonucleotides improve sensitivity of mutation detection based on guanine quenching. AB - Guanine quenching of a fluorescence-labeled DNA probe is a powerful tool for detecting a mutation in a targeted site of a DNA strand. However, a different guanine adjacent to a targeted site can interfere with detection of a point mutation, resulting in unsatisfactory sensitivity. In the current study, we developed a simple method to improve sensitivity of the guanine quenching method using a masking DNA oligonucleotide. The simple addition of a masking DNA oligonucleotide was found to mask the interference of a different guanine in a target oligonucleotide on fluorescence and to enhance difference in the quenching ratio between wild-type and mutant oligonucleotides. Based on this strategy, we succeeded in discriminating various mutations from the wild-type YMDD motif of the hepatitis B virus DNA polymerase gene using guanine quenching with a masking oligonucleotide. PMID- 16701076 TI - The role of subunit epsilon in the catalysis and regulation of FOF1-ATP synthase. AB - The regulation of ATP synthase activity is complex and involves several distinct mechanisms. In bacteria and chloroplasts, subunit epsilon plays an important role in this regulation, (i) affecting the efficiency of coupling, (ii) influencing the catalytic pathway, and (iii) selectively inhibiting ATP hydrolysis activity. Several experimental studies indicate that the regulation is achieved through large conformational transitions of the alpha-helical C-terminal domain of subunit epsilon that occur in response to membrane energization, change in ATP/ADP ratio or addition of inhibitors. This review summarizes the experimental data obtained on different organisms that clarify some basic features as well as some molecular details of this regulatory mechanism. Multiple functions of subunit epsilon, its role in the difference between the catalytic pathways of ATP synthesis and hydrolysis and its influence on the inhibition of ATP hydrolysis by ADP are also discussed. PMID- 16701077 TI - Do children with obesity implicitly identify with sedentariness and fat food? AB - We investigated whether youngsters with obesity (n=39) differed from a control group (n=39) in their self-reported attitudes towards and in their implicit identification with physical activity and food. Self-reported attitudes were assessed using a rating scale; implicit identification was assessed using a self concept Implicit Association Task (IAT). Results revealed a marked group difference on the implicit identification with food: Only youngsters without obesity identified themselves more with non-fat food than with fat food. PMID- 16701078 TI - Cognitive-behavioral therapy for women with lifelong vaginismus: process and prognostic factors. AB - Cognitive-behavioral therapy (CBT) seems an effective treatment of lifelong vaginismus, but mechanisms of action have not yet been established. The present study explored whether the effect of CBT for lifelong vaginismus is mediated by changes in fear of penetration and avoidance behavior, which CBT explicitly aims to alter. A second aim of this study was to predict treatment outcome on the basis of pre-treatment variables. Participants with lifelong vaginismus were allocated at random to a 3-months CBT (n=81) or a waiting-list control condition (n=36). Full vaginal penetration with the penis of the partner constituted the primary outcome measure. Change scores in successful 'non-coital penetrative behavior' and 'fear of coitus' were used to measure the mediating variables. Treatment resulted in an increase of intercourse (outcome), a decrease in fear of coitus, and an enhancement of successful non-coital penetration behavior, compared with no treatment. Outcome (intercourse) was partly mediated by changes in fear of coitus and changes in avoidance behavior. No treatment predictors could be detected in this study sample. It is concluded that techniques such as gradual exposure, aimed at decreasing avoidance behavior and penetration fear, constitute an important avenue of change in the treatment of lifelong vaginismus. PMID- 16701079 TI - The h index and career assessment by numbers. AB - Growing demand to quantify the research output from public funding has tempted funding agencies, promotion committees and employers to treat numerical indices of research output more seriously. So many assessment exercises are now conducted worldwide that traditional peer assessment is threatened. Here, we describe a new citation-based index (Hirsh's h index) and examine several factors that might influence it for ecologists and evolutionary biologists, such as gender, country of residence, subdiscipline and total publication output. We suggest that h is not obviously superior to other indices that rely on citations and publication counts to assess research performance. PMID- 16701080 TI - Female finery is not for males. AB - There has recently been an increase in interest in the notion that female ornamentation is selected through male choice, rather than being an artefact of selection on male ornamentation. There are, however, key differences between the sexes in the type of selection pressures that are likely to generate ornamentation and important differences in investment tradeoffs. Here, I discuss that female ornamentation might be selected more often through female competition over resources than through competition over mates, as exemplified in a recent study by Heinsohn and colleagues. PMID- 16701081 TI - Impacts of global atmospheric change on tropical forests. PMID- 16701083 TI - Rebuilding community ecology from functional traits. AB - There is considerable debate about whether community ecology will ever produce general principles. We suggest here that this can be achieved but that community ecology has lost its way by focusing on pairwise species interactions independent of the environment. We assert that community ecology should return to an emphasis on four themes that are tied together by a two-step process: how the fundamental niche is governed by functional traits within the context of abiotic environmental gradients; and how the interaction between traits and fundamental niches maps onto the realized niche in the context of a biotic interaction milieu. We suggest this approach can create a more quantitative and predictive science that can more readily address issues of global change. PMID- 16701084 TI - From patterns to emerging processes in mechanistic urban ecology. AB - Rapid urbanization has become an area of crucial concern in conservation owing to the radical changes in habitat structure and loss of species engendered by urban and suburban development. Here, we draw on recent mechanistic ecological studies to argue that, in addition to altered habitat structure, three major processes contribute to the patterns of reduced species diversity and elevated abundance of many species in urban environments. These activities, in turn, lead to changes in animal behavior, morphology and genetics, as well as in selection pressures on animals and plants. Thus, the key to understanding urban patterns is to balance studying processes at the individual level with an integrated examination of environmental forces at the ecosystem scale. PMID- 16701085 TI - A distinct urban biogeochemistry? AB - Most of the global human population lives in urban areas where biogeochemical cycles are controlled by complex interactions between society and the environment. Urban ecology is an emerging discipline that seeks to understand these interactions, and one of the grand challenges for urban ecologists is to develop models that encompass the myriad influences of people on biogeochemistry. We suggest here that existing models, developed primarily in unmanaged and agricultural ecosystems, work poorly in urban ecosystems because they do not include human biogeochemical controls such as impervious surface proliferation, engineered aqueous flow paths, landscaping choices, and human demographic trends. Incorporating these human controls into biogeochemical models will advance urban ecology and will require enhanced collaborations with engineers and social scientists. PMID- 16701086 TI - Biological consequences of Chernobyl: 20 years on. AB - The disaster at the Chernobyl nuclear power plant in 1986 released 80 petabecquerel of radioactive caesium, strontium, plutonium and other radioactive isotopes into the atmosphere, polluting 200 000 km(2) of land in Europe. As we discuss here, several studies have since shown associations between high and low levels of radiation and the abundance, distribution, life history and mutation rates of plants and animals. However, this research is the consequence of investment by a few individuals rather than a concerted research effort by the international community, despite the fact that the effects of the disaster are continent-wide. A coordinated international research effort is therefore needed to further investigate the effects of the disaster, knowledge that could be beneficial if there are further nuclear accidents, including the threat of a "dirty bomb". PMID- 16701087 TI - Biological invasions as disruptors of plant reproductive mutualisms. AB - Invasive alien species affect the composition and functioning of invaded ecosystems in many ways, altering ecological interactions that have arisen over evolutionary timescales. Specifically, disruptions to pollination and seed dispersal mutualistic interactions are often documented, although the profound implications of such impacts are not widely recognized. Such disruptions can occur via the introduction of alien pollinators, seed dispersers, herbivores, predators or plants, and we define here the many potential outcomes of each situation. The frequency and circumstances under which each category of mechanisms operates are also poorly known. Most evidence is from population-level studies, and the implications for global biodiversity are difficult to predict. Further insights are needed on the degree of resilience in interaction networks, but the preliminary picture suggests that invasive species frequently cause profound disruptions to plant reproductive mutualisms. PMID- 16701088 TI - Ecological and evolutionary implications of dinosaur feeding behaviour. AB - Dinosaurs had a wide variety of feeding mechanisms that strongly impacted on their ecology and evolution. Here, we show how novel application of technologies borrowed from medicine and engineering, such as CT scanning and Finite Element Analysis, have recently been combined with traditional approaches to result in significant advances in our understanding of dinosaur palaeobiology. Taxon specific studies are providing quantitative data that can be used to generate and test functional hypotheses relating to jaw mechanics and feeding behaviour. In turn, these data form a basis for investigating larger scale patterns of ecological and macroevolutionary change, such as possible coevolutionary interactions and the influence of feeding adaptations on species richness, which are of more general interest to ecologists and evolutionary biologists. PMID- 16701089 TI - A surgeon's guide to the universe. PMID- 16701090 TI - Risk factors for mortality and intra-abdominal complications after pancreatoduodenectomy: multivariate analysis in 300 patients. AB - BACKGROUND: Studies of risk factors after pancreatoduodenectomy are few: some concern restricted populations and others are based on administrative data. METHODS: Multicenter clinical data were collected for 300 patients undergoing pancreatoduodenectomy to determine (by univariate and multivariate analysis) preoperative and intraoperative risk factors for mortality and intra-abdominal complications (IACs), including pancreatic fistula. Fourteen factors including the center and volume effect were analyzed. RESULTS: In univariate analysis, mortality was increased with age 70 years or more, extended resection(s), and volume and center effects. IACs occurred more often with main pancreatic duct diameter of 3 mm or less, normal parenchyma texture, extended resection(s), and the center effect. Pancreatic fistula was more frequent with main pancreatic duct diameter of 3 mm or less, normal parenchyma texture, and the center effect. In multivariate analysis, independent risk factor(s) for mortality were age greater than 70 years (odds ratio [OR], 3; 95% confidence interval [CI], 1.3-8) and extended resection (OR, 5; 95% CI, 1.2-22), risk factors for IACs were extended resection (OR, 5; 95% CI, 1.2-22) and main pancreatic duct diameter of 3 mm or less (OR, 2; 95% CI, 1.1-3), and the risk factor for pancreatic fistula was main pancreatic duct diameter of 3 mm or less (OR, 2.5; 95% CI, 1.2-4.6). CONCLUSIONS: Age more than 70 years, extended resections, and main pancreatic duct diameter less than 3 mm are independent risk factors that should be considered in indications for and techniques of pancreatoduodenectomy. PMID- 16701091 TI - Perioperative complications after hepatectomy with or without intra-arterial chemotherapy for bilobar colorectal cancer liver metastases. AB - BACKGROUND: We investigated perioperative complications of hepatic arterial infusion chemotherapy preceding major hepatectomy for multiple bilobar colorectal cancer metastases. No consensus exists concerning operative feasibility or perioperative course in patients undergoing major liver resection with neoadjuvant chemotherapy--partly because such chemotherapy is considered hepatotoxic, increasing the risk of postoperative liver failure. METHODS: Clinicopathologic data were available for 41 consecutive patients with 5 or more bilobar liver metastases from colorectal cancer who underwent major liver resection with or without prior hepatic arterial chemotherapy. Data concerning operative feasibility, postoperative liver function, complication rates, and histologic findings in the non-neoplastic liver were analyzed retrospectively. RESULTS: Prehepatectomy and postoperative day 1 platelet counts were lower (P < .01 and P < .05), alkaline phosphatase on postoperative day 3 was higher (P < .01), and prothrombin time on day 1 was more prolonged (P < .01) in the chemotherapy group. No significant difference was seen between groups in intraoperative data, morbidity, or duration of hospitalization. Histologic examination of adjacent non-neoplastic liver confirmed mild to severe fatty degeneration in 91% of the patients undergoing neoadjuvant chemotherapy, compared with 53% in those without neoadjuvant chemotherapy (P = .023). Although the number of neoplasms in chemotherapy patients was greater than that of the other group, overall and disease-free survival rates were comparable between groups. CONCLUSIONS: Despite mild postoperative liver dysfunction, pre-resection hepatic arterial chemotherapy did not increase morbidity. PMID- 16701092 TI - True aneurysms of the pancreaticoduodenal artery: successful non-operative management. AB - BACKGROUND: True aneurysms of the pancreaticoduodenal arteries (PDA) are rare, often ruptured, and treated by operation with a high level of mortality. We review our experience since 1994 and that of the literature in the past 20 years to provide management guidelines for this uncommon clinical entity. About 100 cases of PDA aneurysms are described in the literature, most of them as case report. METHODS: Nine patients were admitted to our institution between 1994 and 2004 for true aneurysm of the PDA. They were analyzed with regard to the clinical presentation, radiologic findings, management, and outcome. RESULTS: Seven patients presented for sudden abdominal pain from retroperitoneal hemorrhage. In 2 patients PDA aneurysm was an incidental finding. Abdominal ultrasonography, computed tomographic scan, and visceral angiography was carried out in all cases. Aneurysms ranged from 4 to 30 mm (median, 16.5) in size. Celiac axis stenosis or occlusion was identified in 3 patients. One patient required emergent laparotomy for intra-abdominal rupture of a retro peritoneal hematoma. Therapeutic embolization was successful in all 9 patients. All except 1 are alive with no evidence of recurrence of the true PDA aneurysm with a mean follow-up of 59 months. CONCLUSIONS: The authors recommend definitive treatment of all true aneurysms PDA because of their high risk of rupture. Ruptured PDA aneurysms suspected on CT-scan requires emergent visceral angiography and selective embolization as definitive treatment. PMID- 16701093 TI - Efficacy of accelerated partial breast irradiation as a neoadjuvant treatment for patients with breast cancer: a pilot study. AB - BACKGROUND: Breast conserving treatment (BCT) consists of breast-conserving operation and followed by whole-breast irradiation (WBI). Accelerated partial breast irradiation (APBI) is being considered as a possible alternative to WBI. Neoadjuvant APBI might provide more benefit than postsurgical APBI because tumor downstaging will enhance the likelihood of BCT. METHODS: APBI was delivered as 50 Gy in 5 fractions over 5 days before operation for patients with breast cancer of 3 to 4 cm in diameter. Patients with tumors 3 cm or less were deemed to be candidates for breast-conserving operation. RESULTS: Between September 1998 and August 1999, 12 women were enrolled. The mean tumor diameter and volume were reduced from 3.4 to 1.8 cm (reduction rate: 47%) and from 8.1 to 2.2 cm3 (reduction rate: 71%), respectively. The mean pathologic tumor size was 1.5 cm, and a complete pathologic response was found in 1 patient (8%). All patients were eligible for breast-conserving operation. No ipsilateral breast recurrences have been observed to date. CONCLUSIONS: This is the first report of neoadjuvant APBI for relatively large breast cancers. Although the number of participants was small, these results would encourage the development of clinical trials exploring the efficacy of neoadjuvant APBI. PMID- 16701094 TI - Double mapping with subareolar blue dye and peritumoral green dye injections decreases the false-negative rate of dye-only sentinel node biopsy for early breast cancer: 2-site injection is more accurate than 1-site injection. AB - BACKGROUND: The optimum sentinel node biopsy (SNB) mapping method for breast cancer remains to be determined. No matter which mapping agents are used, 2-site injection may be superior to 1-site injection in limiting the false-negative rate. METHODS: We examined whether a double-mapping method with subareolar injection of blue dye and peritumoral injection of green dye would decrease the false-negative rate of dye-only SNB in 145 patients with early breast cancer. RESULTS: The identification rate for blue-dyed and/or green-dyed (including mixed color-dyed) lymph nodes was 96.6% (140/145). Sensitivity and specificity were 95.1% (39/41) and 100% (99 of 99), respectively. Accuracy was 98.6% (138/140) with a false-negative rate of 4.9% (2/41). There were 4 patients in whom nodes of each color were found, but nodes of only 1 color were shown to be positive. The primary tumors of these 4 patients and of the 2 patients with false-negative results were located in the upper-outer quadrant of the breast. When only blue dyed or green-dyed nodes (including mixed color-dyed nodes) were counted, the false-negative rates were 10.3% (4/39) for the subareolar mapping technique and 10.0% (4/40) for the peritumoral mapping technique. CONCLUSIONS: The double mapping method based on subareolar and peritumoral injections decreases the false negative rate of dye-only SNB for early breast cancer. Variations in lymphatic channels may exist in the lateral half of the breast and thus may influence identification of positive sentinel nodes. This finding should be taken into account in cases of multicentric breast cancer. PMID- 16701095 TI - Sentinel lymph node biopsy for breast cancer: is two-site injection best? PMID- 16701096 TI - Topical Sulfamylon cream inhibits DNA and protein synthesis in the skin donor site wound. AB - BACKGROUND: Whereas Sulfamylon is effective in treatment of burn wound infection, controversy exists regarding its effect on the healing process. METHODS: A partial thickness skin donor site wound was created on the back and indwelling catheters were placed in the carotid artery and jugular vein in rabbits under general anesthesia. Sulfamylon cream (8.5%, BERTEK Pharmaceuticals Inc., Morgantown, W Va) was applied on the wound, with either open or occlusive dressing. The control wound was covered with dressings only. On day 7 after injury, stable isotope tracers were infused to determine the fractional synthetic rate (FSR) of DNA, and FSR and fractional breakdown rate (FBR) of protein in the wound. RESULTS: In the Sulfamylon-open dressing group, the DNA FSR was 1.3 +/- 0.6%/day, the protein FSR was 8.0 +/- 3.5%/day, and the net protein deposition (FSR - FBR) was -0.3 +/- 3.7%/day. These values were lower (P < .01 to .05) than the corresponding values in the control group (DNA FSR: 2.9 +/- 0.9%/day; protein FSR: 20.5 +/- 8.4%/day; net protein deposition: 7.9 +/- 6.0%/day). Sulfamylon cream selectively inhibited DNA FSR from the de novo base synthesis pathway (2.3 +/- 1.2 vs 0.8 +/- 0.5%/day, P < .05 vs control). With the occlusive dressing Sulfamylon cream did not decrease wound DNA FSR due to a stimulation of the base salvage pathway, but still decreased protein FSR (11.5 +/- 5.1%/day, P < .05 vs control). Histologic slides indicated that Sulfamylon cream inhibited re epithelialization, collagen formation, and angiogenesis in the wound. CONCLUSIONS: Topical Sulfamylon cream application inhibited DNA and protein synthesis in the wound, which would be expected to retard the healing process. PMID- 16701097 TI - Is saphenofemoral junction reconstruction necessary during stripping of the saphenous vein? AB - BACKGROUND: Patients who had undergone complete ankle-to-groin stripping of the greater saphenous vein were evaluated retrospectively to assess the necessity of saphenofemoral junction reconstruction during the stripping procedure. Since 1996, in addition to the conventional complete stripping operation, we routinely perform a saphenofemoral junction reconstruction in patients presenting with greater saphenous vein reflux associated with low-grade (grades I-II) saphenofemoral junctional reflux. In this method, the size of the common femoral vein was adjusted to the desired diameter by a running linear suture technique after division of the greater saphenous vein. METHODS: Retrospective evaluation revealed that 73 limbs in 56 patients treated with this technique (group I). This group of patients was matched to another group of 65 patients (78 limbs) with similar characteristics and symptoms (group II) in whom the conventional complete ankle-to-groin stripping of greater saphenous vein was the treatment. The 2 groups were compared with respect to the incidence of complications, including recurrence of varicosities, ecchymosis, lymphocele, lymphorrhagia, wound infection, and paresthesia in the operated extremity. All patients also were evaluated by Doppler ultrasonography at 6 months, 12 months, and annually thereafter to determine the saphenofemoral junction reflux time (valve reflux time). The mean duration +/- SD of follow-up was 6.7 +/- 1.6 years (range, 2.1 10.8 years). RESULTS: Recurrence of varicosity was noted in 14 patients, 3 in group I and 11 in group II (P = .02). There were no statistically significant differences between the 2 groups in terms of ecchymosis, hematoma, lymphocele, lymphorrhagia, wound infection, and paresthesia. At 6 months, a rapid decrease in valve reflux time was noted in group I (P = .0001). In addition, there was a significant improvement in valve reflux time at each subsequent Doppler examination in group I. Group II showed a decrease in valve reflux time, compared with the preoperative value (P = .068). During subsequent Doppler examinations, a decrease in valve reflux time also was noted in group II; this difference reached statistical significance only at 24 months (P = .04). CONCLUSIONS: We believe that saphenofemoral junction reconstruction is a simple technique to perform and that addition of this method to the conventional stripping provides more durable results with a lesser incidence of recurrence. This method should be considered as a treatment modality in patients with greater saphenous vein reflux associated with low-grade (grades I-II) saphenofemoral junctional reflux. PMID- 16701098 TI - Successful 40-hour preservation of the canine small intestine with the cavitary 2 layer method with glutamine supplementation. AB - BACKGROUND: We recently reported that the cavitary 2-layer method (cTLM) allowed stable 24-hour preservation of canine intestine. The aim of this study was to examine the possibility of the 40-hour preservation by cTLM that is supplemented with glutamine. METHODS: Canine jejunal segments (40 cm) were allotransplanted heterotopically without storage (group 1), after 40-hour cold storage with University of Wisconsin solution (group 2), cTLM (group 3), UW with 2% glutamine (group 4), or cTLM with 2% glutamine (group 5). Mucosal glutamine, histidine, tryptophan, glutathione, and adenosine triphosphate concentrations were determined immediately after preservation. At day 7 after the transplantation, maltose absorption test and histopathologic analysis were performed. RESULTS: Mucosal glutamine concentrations increased significantly with glutamine supplementation during preservation in groups 4 and 5 (P < .01). Mucosal adenosine triphosphate levels in cTLM groups (groups 3 and 5) were similar to those in group 1; group 4 showed very low levels after preservation, despite glutamine supplementation. Mucosal glutathione did not differ among groups 2, 3, 4, and 5 and were significantly lower than in group 1. In the absorption test, serum glucose curves showed a peak level by 30 minutes in groups 1 and 5; delayed peaks were seen in groups 2, 3, and 4 compared with group 1. The villous heights were 879, 555, 685, 688, and 773 microm in groups 1, 2, 3, 4, and 5, respectively (group 1 vs group 2, 3, and 4: P < .05). CONCLUSION: cTLM that was supplemented with glutamine extended a safe preservation period up to 40 hours in canine segmental small intestinal transplantation, although this study failed to elucidate a precise mechanism of the glutamine beneficial effect on the graft mucosa. PMID- 16701099 TI - Hepatic ischemia/reperfusion injury is prevented by a novel matrix metalloproteinase inhibitor, ONO-4817. AB - BACKGROUND: Matrix metalloproteinases (MMPs) play an important role in inflammation and neoplastic invasion and metastasis. Little is known about the effects of MMP inhibitors on hepatic ischemia/reperfusion injury. The aim of this study is to examine the inhibitory effects of ONO-4817 (oral inhibitor of MMPs) in rats. METHODS: Hepatic ischemia/reperfusion was induced in male Wister rats by clamping the portal vein and hepatic artery. The animals were randomized into an ONO-4817 group (300 mg/kg body weight per/day) and a vehicle group by oral gavage of a test substance. Serum alanine aminotransferase, histologic changes, gelatinolytic activity, MMP-2 and MMP-9 activities, tissue inhibitor of metalloproteinase 2 (TIMP-2) messenger RNA (mRNA) levels, and mRNA and serum levels of tumor necrosis factor alpha (TNFalpha) and interleukin 1beta (IL-1beta) were measured in both groups. RESULTS: ONO-4817 prevented ischemia/reperfusion injury to the hepatocytes as shown by significant reductions of serum alanine aminotransferase and less severe histologic changes. Gelatinolytic activity was inhibited markedly in the liver of the ONO-4817 group as demonstrated by film in situ zymography. MMP-9 and MMP-2 activities also were inhibited in the ONO-4817 group as shown by gelatin zymography. TIMP-2 mRNA levels showed no significant differences between the 2 groups. TNFalpha mRNA showed no downregulation, but IL 1beta mRNA was downregulated in the liver of the ONO-4817 group 1 to 3 hours after reperfusion. Serum levels of TNFalpha and IL-1beta showed a significant decrease in the ONO-4817 group, compared with the vehicle group after reperfusion. CONCLUSIONS: Hepatic ischemia/reperfusion injury was improved by a novel MMP inhibitor, ONO-4817, not only by inhibition of gelatinolytic activity but also by a decrease in release of inflammatory cytokines. PMID- 16701100 TI - Sonic Hedgehog-dependent proliferation in a series of patients with colorectal cancer. AB - BACKGROUND: The Hedgehog (Hh) gene family is known to regulate development of stem cells. In addition, activation is responsible for the induction of GLI1 proto-oncogene and subsequent cellular proliferation. Sonic Hedgehog (SHh), one of the Hh family members promotes carcinogenesis in airway and pancreatic epithelia, is expressed in colonic stem cells. As differentiated colonic cells arise from constant renewal of Hedgehog-expressing colonic stem cells, SHh could be involved in human colonic carcinogenesis. METHODS: Tissue samples of colorectal adenocarcinoma (T) and adjacent normal colon tissue (NT) were sampled from each of 44 consecutive patients with colorectal cancer. Specific transcription of SHh, GLI1, and the GLI1 downstream target FOXM1 were evaluated using semiquantitative reverse transcriptase polymerase chain reaction. Similar in vitro measurements of mRNA of GLI1 and FOXM1 transcription levels after specific induction by SHh-Np were performed in the HT-29 colorectal tumor cell line to confirm the in vivo results. RESULTS: SHh mRNA was overexpressed in colorectal adenocarcinomas in 38 of 44 (86%) patients. Expression of transcription levels of GLI1 and FOXM1 correlated with SHh expression (SHh vs GLI1, r = 0.77, P < .0001; GLI1 vs FOXM1, r = 0.68, P < .0001; SHh vs FOXM1, r = 0.79, P < .0001). SHh overexpression did not appear to correlate with the patient characteristics evaluated. Similarly, when studied in the HT-29 colorectal cell line, exogenous SHh promoted cell proliferation, while inhibition of SHh expression decreased proliferation. Expression of GLI1 and FOXM1 mRNA increased with exogenous exposure to SHh. CONCLUSIONS: We demonstrated increased expression of SHh mRNA in human colonic adenocarcinomas and in a colorectal cell line with downstream increased expression of GLI1 and FOXM1 mRNA known to promote cell proliferation. This upregulation within human colorectal adenocarcinoma tissue confirms the potential role of the Hh pathway in colorectal carcinogenesis and suggests a potential therapeutic target of Hh blockade in colorectal cancer. PMID- 16701101 TI - Ischemia and prolonged reperfusion before anastomotic construction do not reduce wound strength in the rat intestine. AB - BACKGROUND: Under certain conditions, transient intestinal ischemia can reduce anastomotic strength. Preliminary findings suggest that prolonged reperfusion time, before anastomotic construction, results in reduced wound strength. The purpose of this study is to determine if wound strength indeed decreases with increasing duration of the interval between an ischemic period and construction of an anastomosis. METHODS: In male Wistar rats, ischemia was induced by crossclamping the superior mesenteric artery (SMA) for 40 minutes. In control groups, the SMA was exposed but not clamped. Resection and anastomosis in both ileum and colon were performed immediately after release of the clamp or after 90 minutes or 24 hours. Both the anastomotic bursting pressure and breaking strength were measured after 3 or 5 days, together with hydroxyproline levels. RESULTS: Neither bursting pressure nor breaking strength, either in ileum or in colon, changed significantly when the time between the end of ischemia and anastomotic construction increased. Similar values were obtained in all experimental and corresponding control groups. In the group in which anastomoses were constructed after 24-hour reperfusion, mechanical strength increased significantly from day 3 to day 5 and at the same rate as in the control group. No differences in anastomotic hydroxyproline levels were found between experimental and control groups analyzed at day 5. CONCLUSIONS: A prolonged interval between intestinal ischemia and anastomotic construction does not affect development of early wound strength. Therefore, delayed anastomosis after transient ischemia is not likely to increase the risk of anastomotic complications. PMID- 16701103 TI - Hemodynamic and metabolic effects of selective beta1 adrenergic blockade during sepsis. AB - BACKGROUND: Selective beta adrenergic antagonists are commonly used to reduce myocardial demise in patients at risk for cardiac-related death. The purpose of this study was to examine the hemodynamic and metabolic effects of cardiac selective beta adrenergic blockade in patients. METHODS: Muscle protein kinetics were quantified using isotopic tracer methodology in 6 moderately septic, mechanically ventilated patients with pneumonia before and then at the conclusion of a 3-hour infusion of esmolol of sufficient dose to reduce heart rate by 20% from baseline. A battery of hemodynamic measurements as facilitated by a thermodilution pulmonary artery catheter and indirect calorimetry were also measured before and after the 3-hour selective beta adrenergic blockade. RESULTS: Selective beta adrenergic blockade was associated with the 20% reduction in heart rate and a comparable decrease in cardiac output. Esmolol administration failed to affect systemic or pulmonary vascular resistance, oxygen consumption, hepatic or leg blood flow, energy expenditure, or ATP availability/energy charge within muscle. Esmolol infuse did incite a shift in fuel oxidation toward an increase in palmitate oxidation and with a decrease in the oxidation of glucose. There was no demonstrable influence beta1 adrenergic blockade on muscle protein kinetics. CONCLUSIONS: Cardiac selective beta adrenergic blockade with esmolol reduces cardiac output in proportion to the percentage decreases in heart rate in moderately severe septic patients without adversely affecting oxygen utilization or hepatic, peripheral blood flow. PMID- 16701102 TI - Squamous cell carcinoma-antigen messenger RNA level in peripheral blood predicts recurrence after resection in patients with esophageal squamous cell carcinoma. AB - BACKGROUND: The aim of this study was to clarify whether preoperative squamous cell carcinoma-antigen messenger RNA (SCC-Ag mRNA) level in peripheral blood can be used to predict tumor recurrence after curative resection for esophageal squamous cell carcinoma. METHODS: A prospective analysis was conducted for 46 consecutive patients who underwent curative esophagectomy and who had no residual tumor. The SCC-Ag mRNA level in the peripheral blood of each patient was measured preoperatively by using quantitative reverse transcriptase-polymerase chain reaction. Median follow-up period was 34 months. RESULTS: Receiver operating characteristic analysis demonstrated that the optimal cutoff level of SCC-Ag mRNA was 40. Patients were divided into the high SCC-Ag mRNA level group (n = 14) and the low SCC-Ag mRNA level group (n = 32). The cumulative probabilities of tumor recurrence were higher in the high SCC-Ag mRNA level group (probability of recurrence was 71% at 2 years) than in the low group (22% at 2 years; P = .0005). SCC-Ag mRNA level (relative risk, 3.00; 95% confidence interval, 1.05-8.54; P = .040) was the strongest independent predictor of recurrence by multivariate analysis. CONCLUSIONS: Preoperative SCC-Ag mRNA levels in the peripheral blood are the best predictive factor for recurrence in patients with esophageal squamous cell carcinoma who undergo curative resection (R0). PMID- 16701105 TI - Clinical factors used to support prophylactic mastectomy for a male BRCA2 mutation carrier. PMID- 16701104 TI - Video-assisted living donor hemihepatectomy through a 12-cm incision for adult-to adult liver transplantation. AB - OBJECTIVES: There has been remarkable progress in recent technical innovations for laparoscopic hepatectomy. However, a laparoscopic procedure rarely has been indicated for donation of the liver in living-related liver transplantation (LRLT). Here, we described the technique and the outcome of video-assisted donor hepatectomy (VADH) for adult-to-adult LRLT. METHODS: For 13 donors in adult-to adult LRLT, 3 types of major hepatectomy--right hemihepatectomy (3), and left hemihepatectomy, with or without the caudate lobe (10)--were performed through video-assisted procedures; surgical manipulation via ports or via a 12-cm incision and viewing through a laparoscope or through incision were combined and used. RESULTS: VADH was completed in 13 donors, with a median operation time of 363 +/- 33 minutes and a median blood loss of 302 +/- 191 mL. No complications specific to video-assisted procedures, postoperative bile leak, or bleeding were observed. The restoration of the liver function was smooth, and the use of an analgesic (median: 1.2 times) was reduced, compared with the historical control (median: 3.8 times) that underwent a standard donation of the liver. Currently, all donors are healthy and have returned to their previous activities. The grafts have been functioning well, excluding 3 recipients who succumbed to serious complications unrelated to the video-assisted procedure. CONCLUSION: We have shown a new method of VADH through a 12-cm laparotomy for adult-to-adult LRLT. This technique is as feasible as standard open donor hepatectomy, with less pain and with improved postoperative symptoms. PMID- 16701106 TI - Images in surgery. Tuberculosis peritonitis. PMID- 16701107 TI - Advanced endometrial cancer: is lymphadenectomy necessary or sufficient? PMID- 16701108 TI - Racial disparities research: it's not just black and white. PMID- 16701109 TI - Nursing intervention and smoking cessation: meta-analysis update. AB - OBJECTIVE: The study objective was to determine through meta-analysis the effects of nursing-delivered smoking-cessation interventions. RESULTS: Thirty-four studies met inclusion criteria in this updated meta-analysis. Twenty-six studies compared a nursing intervention with a control or usual care group of adults and found interventions of high and low intensity to modestly increase the odds of quitting (1.36, 95% confidence interval 1.22-1.51). The study results demonstrated heterogeneity; using a random effects model did not make a difference. There was evidence that interventions were most effective for hospital inpatients with cardiovascular disease than for patients with other conditions (odds ratio 2.14, confidence interval 1.39-3.31). Interventions in nonhospitalized adults were beneficial as well; no effect was found for additive intervention components. Counseling during health-screening programs or as part of multifactorial secondary preventions programs was found to be the least effective. The challenge will be to incorporate smoking-cessation interventions into evidence-based nursing practice. PMID- 16701110 TI - Public knowledge of heart attack in a Nepalese population survey. AB - OBJECTIVE: Limited knowledge of heart attack symptoms may prevent patients from seeking time-dependent thrombolytic therapy, an intervention that offers impressive survival benefit. Previous studies carried out in developed countries demonstrated a deficit of knowledge about a wide range of heart attack symptoms. The aim of this study was to describe knowledge of heart attack, knowledge of heart attack symptoms, and anticipated first response to symptoms among the lay public in Nepal. METHODS: A total of 1192 participants (657 men and 535 women age 16 to 88 years old) were interviewed in a cross-sectional manner. Those <16 years of age, all health professionals, and individuals with a history of heart attack were excluded. RESULTS: A total of 862 (72.3%) participants had heard of heart attack. Significantly more male than female participants had heard of heart attack (P <.001). Of the respondents, 91.7% with >or=10 years of education (ED-2) had heard of heart of attack, whereas only 54% respondents with <10 years of education or who were illiterate (unable to read and write) (ED-1) had heard of heart attack, and in both the male and female populations, a higher percentage of the ED-2 group had heard of heart attack than the ED-1 group (92.6% vs. 60% and 85.6% vs. 49.6%, respectively). A significantly higher number of respondents from 31 to 50 years of age (AGE-2) had heard of heart attack than those 16 to 30 years of age (AGE-1) and those >50 years of age (AGE-3) (P <.001). Among 862 respondents who had heard of heart attack, 21.3% could not name any heart attack symptoms. A total of 16 different heart attack symptoms were named. Fainting or collapsing (48%), chest pain (22.4%), shortness of breath (9%), dizziness (8.4%), palpitations (7.4%), and sweating (7.4%) were the leading symptoms named by respondents. Fainting or collapsing and chest pain and shortness of breath were named more frequently among the ED-2 group respondents and the AGE-3 group men. Only 3.7% could name >or=2 typical heart attack symptoms. A significantly larger number of the ED-2 group named >or=2 typical symptoms than their counterparts (P <0.001). A large number (77.6%) of respondents preferred immediate hospital referral and/or doctor consultation after a heart attack. CONCLUSIONS: In Nepal, better-educated men are more aware of heart attack. Fainting or collapsing and chest pain and shortness of breath were leading heart attack symptoms named by the general population. Public heart attack awareness is not adequate and knowledge of wide range of heart attack symptoms is deficient in the Nepalese general population. PMID- 16701111 TI - Relation of atrial arrhythmia-related symptoms to health-related quality of life in patients with newly diagnosed atrial fibrillation: a community hospital-based cohort. AB - BACKGROUND: Atrial fibrillation is the most common sustained cardiac rhythm disturbance encountered in clinical practice. Despite its high prevalence and associated hospital admissions, few studies document the actual perceived symptoms of atrial fibrillation and its relationship to health-related quality of life. OBJECTIVES: The purposes of this study were to explore the patients' symptoms of atrial fibrillation and to examine the relationship between specific symptoms and self-reported health-related quality of life in patients newly diagnosed with atrial fibrillation. METHODS: This study used a descriptive correlational and cross-sectional survey design using a face-to-face interview method. Arrhythmia-related symptoms and health-related quality of life were measured by Version 3 of the Symptom Checklists: Frequency and Severity and the Short Form-36 Health Survey of the Medical Outcomes Study questionnaire, respectively. The data collected were statistically analyzed by t tests and partial correlation analyses to examine the group differences in symptoms and the relationship between arrhythmia-related symptoms and health-related quality of life, respectively. RESULTS: A total of 81 subjects newly diagnosed with atrial fibrillation within 6 months before data collection were interviewed from two community hospitals in Cleveland, Ohio. Among 16 atrial arrhythmia-related symptoms, tiredness was the most frequently reported. Furthermore, reported symptoms were significantly different between the two groups divided only by age. Symptom frequency had a significant correlation with both physical health and mental health. Specifically, the relationships between "shortness of breath" and physical health, and "tiredness" and mental health were shown to be the strongest in each dimension of health-related quality of life. CONCLUSION: This study demonstrates that patients with more frequent symptoms perceived poorer physical health and mental health compared with those with less frequent symptoms. Thus, the general conception that "atrial fibrillation is asymptomatic" should be reconsidered. In clinical situations, as a health care provider caring for patients with atrial fibrillation, the assessment of the nature and severity of symptoms should be performed to promote patients' quality of life. PMID- 16701112 TI - Functional recovery after neuromuscular blockade in mechanically ventilated critically ill patients. AB - BACKGROUND: An estimated 24% to 70% of individuals have prolonged paralysis or severe weakness after receiving neuromuscular blocking agents (NMBAs) when therapy is terminated. OBJECTIVES: The purposes of this study were to (1) evaluate the relationship between recovery of neuromuscular transmission (NMT) and functional muscle activity after NMBA administration; (2) evaluate the relationship between delayed recovery of NMT or muscle activity and functional performance; and (3) determine the predictors of delayed recovery of NMT, muscle activity, and functional performance. METHODS: This was a multisite study using a prospective, nonexperimental, descriptive design with convenience sampling techniques. Instruments used included a five-point muscle score, Actigraph, and peripheral nerve stimulator. RESULTS: Key findings were as follows: (1) NMT returned promptly, whereas muscle activity remained severely depressed; (2) only two subjects (5%) recovered functional performance within 24 hours; (3) degree of muscle weakness immediately after neuromuscular blockade was associated with prolonged time to extubation and mobility; and (4) predictors of delayed recovery included cumulative dose of aminosteroid NMBAs, age, and renal function. CONCLUSION: Prolonged recovery of muscle activity and extreme weakness may occur despite brisk recovery of NMT after neuromuscular blockade. PMID- 16701113 TI - Caring for organ donors: the intensive care unit physicians' view. AB - This study examined the meanings that the intensive care unit (ICU) physicians attribute to their practice when caring for brain-dead organ donors. It is a phenomenological study, a qualitative method that searches for describing and understanding the experiences lived. Data were collected through recorded individual interviews made with 10 ICU physicians who work in a university hospital in the interior of Sao Paulo, Brazil. Three categories emerged from data: (1) providing care for brain-dead organ donors; (2) relating to donors and their families; and (3) the ethical concerns and self-awareness of the physicians. There is consensus as to many aspects: demand of technical qualification and excellence in practice; need of investing in technology of organ transplant; and donors seen as human beings and holders of human rights. Participants understand that family participation is decisive in the donation process, although interactions between the family members and the physicians are difficult because of the sensitive nature of the situation. The physicians often felt ill-prepared to openly discuss the topic of brain death and organ donation. Educational programs for physicians and family members may assist with this difficult process. PMID- 16701114 TI - Critical care nurses' responses to patient photographs displayed at the bedside. AB - Photographs of patients placed at the bedside by family or friends are a common sight in the intensive care unit. Few studies have shown that displaying patient photographs can have the following effects: motivate staff in caring for the patient by creating a reminder of the patient's preillness state; enhance the empathetic bond between nurse and patient; personalize care in a highly technical environment; and improve the communication between the nurse, patient, and family. PMID- 16701115 TI - When brown and blue make red: a case of acquired methemoglobinemia. PMID- 16701116 TI - Pacemaker-induced Staphylococcus aureus mitral valve acute bacterial endocarditis complicated by persistent bacteremia from a coronary stent: Cure with prolonged/high-dose daptomycin without toxicity. AB - Continuous high-grade Staphylococcus aureus bacteremia suggests acute bacterial endocarditis (ABE), a protected focus, ie, an abscess, or a device-related infection. Daptomycin was curative of S. aureus ABE and coronary stent-related bacteremia. Prolonged high-dose daptomycin therapy (12 mg/kg per day for 41 days) is not associated with any toxicity. Persistent S. aureus bacteremia in ABE should suggest myocardial or perivalvular abscess. If intracardiac abscess can be ruled out and there is no extracardiac source of the S. aureus bacteremia, then a device-related infection should be considered. PMID- 16701117 TI - Isolated atrial infarction. PMID- 16701118 TI - Provoking comments and neurobiologic interest in our clinical research study. PMID- 16701119 TI - Can ultrasound help to define orthopedic surgical complications? AB - This article aims to describe and illustrate the usefulness of ultrasound in detecting complications of orthopedic implants, metal fixation devices, and other surgical material, with an emphasis on soft-tissue pathology. PMID- 16701120 TI - Imaging primary and secondary tumor thrombus of the inferior vena cava: multi detector computed tomography and magnetic resonance imaging. AB - Tumor thrombus of the inferior vena cava (IVC) is a severe medical condition with very poor prognosis unless the patient is treated with surgical resection. It can be caused by a primary leiomyosarcoma originating in the vessel wall or by intraluminal extension of tumor thrombus into the IVC from an adjacent organ. We reviewed 21 cases of tumoral thrombus in the IVC including primary leiomyosarcoma of the IVC (2 cases), renal cell carcinoma (14 cases), adrenocortical carcinoma (2 cases), primary adrenocortical leiomyosarcoma (1 case), hepatocellular carcinoma (1 case), and retroperitoneal metastasis (1 case). The most common findings of IVC tumor thrombus by multi-detector CT and magnetic resonance imaging will be discussed, including scanning protocols and the advantages and disadvantages of each method. PMID- 16701121 TI - Multidetector row computed tomography evaluation of potential living laparoscopic renal donors: the story so far. AB - Renal transplantation is the treatment of choice for end-stage renal disease. Living related kidney donation is the major source of renal grafts due to limited availability of cadaveric kidneys. Open nephrectomy was used to harvest donor kidneys. However, the laparoscopic approach is associated with less postoperative pain and quick recovery. So, most centers now prefer a laparoscopic approach to explant donor kidneys. Laparoscopic approach is technically challenging due to limited operative visibility. Hence, accurate preoperative detection of renal arterial and venous anomalies is imperative to avoid inadvertent vascular injury and bleeding. The preoperative workup of renal donors includes clinical evaluation, laboratory tests, and imaging. Traditionally, the renal donors were evaluated with conventional imaging techniques, which included renal catheter angiography and intravenous urography. However, conventional imaging is invasive, expensive, and less accurate for evaluation of complex renal venous anomalies, small calculi, and diffuse or focal renal parenchymal lesions. The introduction of multidetector row computed tomography (MDCT) revolutionized the CT technology by enabling isotropic resolution with faster scan coverage in a single, short breath-hold. Consequently, MDCT has now replaced conventional imaging for comprehensive imaging of potential living renal donors. MDCT is a minimally invasive technique that can accurately detect urolithiasis, renal arterial and venous anomalies, renal parenchymal lesions, and urinary tract anomalies. Renal vascular anomalies detected by MDCT can help the surgeon in planning donor nephrectomy. MDCT with three-dimensional CT angiography enables accurate preoperative renal vascular mapping. This article reviews the role of MDCT in preoperative evaluation of potential laparoscopic renal donors. PMID- 16701122 TI - Multiple focal nodular hyperplasia: atypical imaging characteristics in the setting of hepatic steatosis. AB - Focal nodular hyperplasia is an uncommon, benign lesion of the liver most often affecting women of childbearing age. It can present clinically with chronic, intermittent right upper quadrant pain and nausea, with associated liver enzyme abnormalities. The presence of characteristic sonographic and computed tomography imaging findings can help to differentiate this entity from other hepatic masses. This article describes an unusual case of multifocal disease that demonstrates atypical imaging findings in the background of hepatic steatosis. PMID- 16701123 TI - Cavernous angioma of the cerebellopontine angle. AB - Although cavernous hemangiomas are commonly found supratentorially, their presence in the internal auditory canal is uncommon. We describe a case that was initially mistaken for a vestibular schwannoma due to its location in the cerebellopontine angle. Although there can be overlap in their presentations, familiarity with the characteristic magnetic resonance imaging findings of both entities will aid in their differentiation and potentially affect preoperative planning. PMID- 16701125 TI - Thumb by metacarpal extension osteotomy: rationale and efficacy for Eaton Stage I disease. AB - This article describes the rationale and results of a "biomechanical" strategy to restore trapeziometacarpal (TM) stability when symptomatic Eaton Stage I disease exists. Though the author has performed TM arthroscopy, synovectomy, and capsular shrinkage for such cases in 10 patients, the author has been dissatisfied with the outcomes, particularly pain relief. The author currently relies exclusively on extension osteotomy as treatment for this subset of patients. Thumb metacarpal extension osteotomy remains an effective treatment alternative for the hypermobile TM joint consistent with Eaton Stage I disease. This procedure alters forces, shifts load away from the volar compartment, and further engages the dorsoradial ligament. Clinical outcomes are favorable, and no bridges are burned should arthritic changes develop in the future. PMID- 16701126 TI - Ligament reconstruction. AB - Volar ligament reconstruction is an effective technique for treating symptomatic laxity of the CMC joint of the thumb. The laxity may bea manifestation of generalized ligament laxity,post-traumatic, or metabolic (Ehler-Danlos). There construction reduces the shear forces on the joint that contribute to the development and persistence of inflammation. Although there have been only a few reports of the results of volar ligament reconstruction, the use of the procedure to treat Stage I and Stage II disease gives good to excellent results consistently. More advanced stages of disease are best treated by trapeziectomy, with or without ligament reconstruction. PMID- 16701127 TI - Trapeziometacarpal arthroscopy: a classification and treatment algorithm. AB - Arthroscopic assessment of the CMC joint allows direct visualization of all components of the joint, including synovium, articular surfaces, ligaments, and the joint capsule. It also allows for the extent of joint pathology to be evaluated and staged with intraoperative management decisions made based on this information. The author recommends this arthroscopic staging to ensure better judgment of this condition in order to provide the most adequate treatment option to patients who have this disabling condition. Future studies assessing the clinical long term results using arthroscopy will likely ensure its place in the treatment armamentarium for trapeziometacarpal osteoarthritis. PMID- 16701128 TI - Trapeziectomy. AB - Primary osteoarthritis of the carpometacarpal joint of the thumb is common, especially in women aged 60 or older. Patients usually present with activity related pain at the thumb base. First treatment may include activity modification, pain relieving medications, splinting, and possibly corticosteroid injections. When these measures fail to pre-serve or restore the patient's quality of life, surgical intervention may be appropriate. Many surgical alternatives are described for the treatment of thumb carpometacarpal joint arthritis, and most begin with at least partial trapeziectomy. Hematoma distraction arthroplasty results in improved outcomes as compared with historical results following trapeziectomy alone. Temporary distraction allows the body's healing response to fill in the trapezial void with scar tissue, obviating the need for ligament reconstruction or tissue interposition. PMID- 16701129 TI - Suspensionplasty for basal joint arthritis: why and how. AB - Abductor pollicis longus suspensionplasty is a simple, effective treatment alternative forbasal joint arthritis. Use of a suspensionplasty technique acknowledges our current understanding of forces involved during pinch and grip, as well as the role of normal ligamentous anatomy. The primary rationale for performing suspensionplasty revolves around resisting the sagittal plane collapse that will occur when the thumb is loaded during pinch. In the absence of a volar based suspension of the metacarpal, cantilever bending forces and axial force transmission will result in the dissipation of force along the thumb lever arm, and ultimately longitudinal collapse. Maximal grip and pinch strength require suspensionplasty, which can be performed using a variety of techniques. The author's current technique for suspensionplasty is described. PMID- 16701130 TI - Rheumatoid arthritis: silicone metacarpophalangeal joint arthroplasty indications, technique, and outcomes. AB - Silicone implant arthroplasty has been used for more than 40 years for severe rheumatoid disease at the metacarpophalangeal (MCP) joint. Multiple investigations have shown that silicone arthroplasty places the MCP joint in a more extended posture, with some improvement in the total arc of motion. Ulnar drift is also improved, but strength and other objective measures have not demonstrated marked changes postoperatively. The lack of prospective data and more complete outcome assessment has been, at least in part, responsible for the marked difference in opinions between rheumatologists and hand surgeons on the effectiveness of MCP arthroplasty. Recent reports using patient-centered outcome measures have shown that early outcome is favorable, with improvements in appearance, pain, and function. PMID- 16701131 TI - Nonrheumatoid metacarpophalangeal joint arthritis. Unconstrained pyrolytic carbon implants: indications, technique, and outcomes. AB - Unconstrained pyrolytic surface replacement arthroplasty provides the benefits of a more natural center of rotation with preservation of native ligamentous joint stability. Initial short-term results show excellent motion, pain relief, and restoration of pinch and grip strength. These results are encouraging, and suggest that pyrolytic carbon arthroplasty may be a reasonable option for joint salvage in patients suffering from MP joint osteoarthritis. PMID- 16701132 TI - Finger metacarpophalangeal joint disease: the role of resection arthroplasty and arthrodesis. AB - When finger MP joint arthrosis exists, it is indeed infrequent that implant arthroplasty is not the most optimal treatment alternative. When post-traumatic bone loss or postinfectious dysfunction require surgical intervention, however, the hand surgeon may need to consider the options of resection arthroplasty and arthrodesis. So long as the MP joint is pain-free and relatively stable, most patterns of functional prehension can be maintained. PMID- 16701133 TI - Prosthetic replacement of the proximal interphalangeal joint. AB - The first proximal interphalangeal joint (PIPJ) replacements were hinged devices allow-ing only single-axis motion. Newer implant arthroplasties of the PIPJ have anatomically designed proximal phalangeal and middle phalangeal components. Constrained devices have typically lead to failure at the hinge mechanism or the prosthetic bone interface. A need to create balanced forces across the joint was the rationale behind the semicon-strained PIPJ prosthesis, which uses an anatomic design. Limitations of the silicone PIPJ implant include its lack of resistance to valgus loading at the index and long digits during the pinch maneuver. It is generally believed that a PIPJ surface replacement arthro-plasty that preserves the collateral ligaments will achieve greater PIPJ stability. PMID- 16701134 TI - Distal interphalangeal joint arthrodesis with screw fixation: why and how. AB - Though DIP joint fusion can be successfully achieved with K-wires in both the osteoarthritic and rheumatoid patient, their use is often some-what of an inconvenience to the patient. They prohibit showering, may become infected, may back out and catch on clothing, and surely slowdown mobilization of the rest of the finger [1]. For optimal prehension, a modest amount of DIP joint flexion is required, however. Thus, one advantage of K-wires is that they allow fusion in 5 degrees to 10 degrees of flexion (Fig. 1). In the rheumatoid patient in particular, bone stock may be so com-promised that getting enough purchase with wires alone can be challenging. Since making the transition to the Herbert screw, hardware-related complications and patient dissatisfaction with obligatory postoperative functional limitations until union is achieved have been eliminated. Despite the fact that the fusion must occur without flexion-a necessity to ensure intramedullary placement of the screw-patients seem to adapt well (Fig. 2). One further potential disadvantage of screw fixation is the issue of size mismatch between phalanx and screw-especially in the small finger. Though cautious insertion is justified, precise technique allows use even in the small finger-a benefit when early motion is indicated; for example, when concomitant proximal interphalangeal (PIP) implant arthroplasty is performed in an adjacent digit. This device is contraindicated, obviously, if future PIP joint arthro plasty is anticipated in the same finger (Fig. 3). PMID- 16701135 TI - Thumb metacarpophalangeal arthritis: arthroplasty or fusion? AB - The human thumb assumes 50% of the workload of the human hand, and is therefore the most important digit. As such, the thumb has a propensity for the development of osteoarthritis. Moreover, the thumb is also often diseased, in anywhere from 68% to 80%of patients who have rheumatoid arthritis. Much attention over the years has been given to the carpalmetacarpal joint of the thumb, whereas the metacarpophalangeal (MP) joint of the thumb remains largely unstudied. The purpose of this article is to review the etiology of thumb MP joint arthritis, and discuss the different treatment options of this condition. PMID- 16701136 TI - Degenerative and post-traumatic arthritis affecting the carpometacarpal joints of the fingers. AB - Symptomatic post-traumatic arthritis affecting the finger CMC joints is less common than might otherwise be expected based on the frequency of injury, especially to the fifth CMC joint. For the fifth CMC joint, the shallow concavoconvex articulation combined with the typical fracture location may provide a protective effect. Non-operative measures are typically successful, except in cases of missed fracture/dislocations and symptomatic joint instability. In these instances, re-construction emphasizes stability first, with an eye toward mobility for the ulnar column. It is common to detect a bony prominence in the region of the dorsal second and third CMC joints during examination of the hand. In most instances, the projection is asymptomatic and likely represents an os styloideum [18]. When painful and unresponsive to nonoperative treatments, this carpometacarpal boss can be excised surgically. PMID- 16701138 TI - Future of allergen-specific immunotherapy: better understanding of the mechanisms, novel treatments, and long-term cure. PMID- 16701139 TI - Allergy diagnosis, allergen repertoires, and their implications for allergen specific immunotherapy. AB - Immunotherapy for allergic diseases represents an important but largely unmet medical need. Conventional immunotherapy suffers from several breakdowns related to the quality of the extracts used, the risk of inducing anaphylactic reactions, and the extremely long treatment time. Many of the problems associated with using natural allergenic products for allergy diagnosis and treatment can be overcome using genetically engineered recombinant allergens. New therapeutic strategies based on recombinant technology include peptide-based vaccines, engineered hypoallergens with reduced IgE-binding properties, nucleotide-conjugated vaccines that promote Th1 responses, and the possibility of developing prophylactic allergen vaccines. PMID- 16701140 TI - Standardization of allergen-specific immunotherapy vaccines. AB - In the 1970s and 1980s, scientific methods were introduced in the standardization of allergen vaccines and, in combination with improved documentation of the clinical benefits obtained using standardized vaccines, specific allergy treatment as a scientifically based, reproducible, and safe treatment for allergic disease was established. This article describes important issues in the control of source materials and vaccine preparation as part of the European standardization of allergen vaccines, and also includes a discussion of vaccines that are based on recombinant allergens, which may appear on the market in the near future. PMID- 16701141 TI - Mechanisms of allergen-specific immunotherapy: T-regulatory cells and more. AB - Activation-induced cell death, anergy, or immune response modulation by regulatory T cells (Treg cells) are essential mechanisms of peripheral T-cell tolerance. Genetic predisposition and environmental instructions tune thresholds for the activation of T cells, other inflammatory cells, and resident tissue cells in allergic diseases. Skewing allergen-specific effector T cells to a Treg cell phenotype seems to be crucial in maintaining a healthy immune response to allergens and successful allergen-specific immunotherapy. The Treg-cell response is characterized by an abolished allergen-specific T-cell proliferation and the suppressed secretion of T-helper 1- and T-helper 2-type cytokines. Suppressed proliferative and cytokine responses against allergens are induced by multiple suppressor factors, including cytokines such as interleukin-10 (IL-10) and transforming growth factor beta (TGF-beta), and cell surface molecules such as cytotoxic T-lymphocyte antigen-4, programmed death-1, and histamine receptor 2. The increased levels of IL-10 and TGF-beta produced by Treg cells potently suppress IgE production while simultaneously increasing the production of noninflammatory isotypes IgG4 and IgA, respectively. In addition, Treg cells directly or indirectly suppress the activity of effector cells of allergic inflammation, such as mast cells, basophils, and eosinophils. In conclusion, peripheral tolerance to allergens is controlled by multiple active suppression mechanisms on T cells, regulation of antibody isotypes, and suppression of effector cells. The application of current knowledge of Treg cells and related mechanisms of peripheral tolerance may soon lead to more rational and safer approaches to the prevention and cure of allergic disease. PMID- 16701142 TI - The role of TGF-beta in allergic inflammation. AB - The transforming growth factor beta (TGF-beta) plays a dual role in allergic disease. It is important in suppressing T cells and also mediates repair responses that lead to unwanted remodeling of tissues. Advances in the immunology of allergy indicate that allergens cause overreactions in the lymphocyte compartment because of the lack or decreased number of suppressive, regulatory T cells. TGF-beta was shown to induce regulatory T cells and participate directly in suppression of effector T cells. Therefore, TGF-beta may help return reactivity to allergens to normal subsymptomatic activity. Whether chronic inflammatory diseases such as asthma profit from TGF-beta-mediated suppression of specific immune responses or whether the TGF-beta-mediated tissue remodeling aggravates diseases more than it helps control immune reactions is unclear. This article addresses these issues and future strategies in this field. PMID- 16701143 TI - Histamine receptors in immune regulation and allergen-specific immunotherapy. AB - The cells involved in the regulation of immune responses and hematopoiesis express histamine receptors and secrete histamine. Histamine acting through four types of its receptors has been shown not only to affect chronic inflammatory responses but also to regulate several essential events in the immune response. Histamine signals have a role in the mechanisms of tolerance induced during allergen-specific immunotherapy (SIT), acting mainly through its receptor (HR) type 2. It positively interferes with the peripheral antigen tolerance induced by T regulatory cells in several pathways. The rationale for the concomitant use of H1 antihistamines during SIT is diverse and includes reduction of its immediate side effects as well as enhancement of mechanisms of specific tolerance and anti inflammatory effects of vaccination. PMID- 16701144 TI - Strategies for recombinant allergen vaccines and fruitful results from first clinical studies. AB - Recombinant DNA technology has delivered the prospect of a new generation of preparations for allergen-specific immunotherapy. The first clinical studies with recombinant allergens have yielded encouraging results, suggesting that there is a good chance that such preparations will become available for use in the routine management of allergic disease. PMID- 16701145 TI - Novel ways for immune intervention in immunotherapy: mucosal allergy vaccines. AB - Allergen-specific immunotherapy is currently the only curative treatment for allergy. Subcutaneous immunotherapy (SCIT) has been successfully used to treat patients who are allergic to insect venom, house dust mites, or tree or grass pollens. In the context of potentially severe, albeit infrequent, side effects associated with SCIT, mucosal routes of administration are being investigated to conduct allergenic desensitization. This article reviews recent developments in the field of nasal, oral, and sublingual immunotherapy as they relate to safety, clinical efficacy, and immune mechanisms of action. Implications for the design and development of improved allergy vaccines that could be used through such nonparenteral routes are discussed. Specifically, allergen presentation platforms and adjuvants facilitating the targeting of immune cells at mucosal surfaces to promote tolerance induction are reviewed. PMID- 16701146 TI - Targeting dendritic cells in allergen immunotherapy. AB - Allergen immunotherapy is a well-established strategy for treating allergic diseases with the goal of inducing allergen-specific tolerance. Identified mechanisms contributing to the therapeutic effect of immunotherapy include a shift of T helper 2 (Th2)-type immune responses to a modified Th2 immune response, a change of the balance of IgE-producing B cells to the production of IgG subtypes, in addition to increased IL-10 and TGF-beta secretion and activation of the suppressive functions of regulatory T-cells. Dendritic cells (DCs), which as outposts of the immune system are capable of T-cell priming through efficient allergen uptake by IgE receptors expressed on their cell surface. Most of the hypotheses concerning the function of DCs as facilitators of allergen-specific tolerance in allergen immunotherapy remain speculative. Therefore, studies must focus on the functional changes of DCs under immunotherapy to close the gap of knowledge about their exact role. These experimental data should help confirm the hypothesis of DCs as efficient silencers and potential target cells and take advantage of the bivalent character and tolerogenic properties of DCs. PMID- 16701147 TI - Peptide immunotherapy. AB - Synthetic peptides representing T-cell epitopes of allergens and autoantigens have been employed to induce antigen-specific tolerance in vivo in experimental models and the clinical setting. Delivery of peptides orally or by injection leads to reduced reactivity to antigen accompanied by the induction of T cells with a regulatory phenotype. Peptide therapy may provide a safe, effective, and economically viable approach for disease-modifying therapy in autoimmune and allergic diseases. PMID- 16701148 TI - IgE-facilitated antigen presentation: role in allergy and the influence of allergen immunotherapy. AB - IgE-facilitated allergen presentation (FAP) is an important pathogenic mechanism in allergic disease and represents a potential therapeutic target. Allergen immunotherapy is a highly effective therapy, particularly in patients with seasonal pollinosis who fail to respond to usual pharmacotherapy. Allergen immunotherapy induces "blocking" IgG antibodies that are detectable in serum and have been shown to inhibit IgE-FAP in vitro. This review summarizes the main components involved in IgE-FAP and the potential value of a validated functional assay of serum inhibitory antibodies for IgE-FAP for monitoring the clinical response to immunotherapy. PMID- 16701149 TI - Biodegradable PLGA particles for improved systemic and mucosal treatment of Type I allergy. AB - Although allergen immunotherapy is basically a story of success, it still needs improvement. The goal of this study was to optimize parenteral and oral allergen formulations through using the biocompatible polymer of lactic and glycolic acid (PLGA). Subcutaneous application of birch pollen allergen Bet v 1 encapsulated in nanoparticles biased the immune response toward Th1 in allergic mice and did not elicit granuloma formation in mice and in human volunteers. When oral immunotherapy of mice was tried with birch pollen-filled PLGA microparticles, mucosal targeting was indispensable for achieving any immune response, and targeting of M-cells was necessary for modulating an ongoing allergic response toward Th1. The authors suggest that biocompatible PLGA nano- or microparticles can be useful tools for upgrading therapy of type I allergy. PMID- 16701150 TI - Bacillus Calmette-Guerin, Mycobacterium bovis, as an immunomodulator in atopic diseases. AB - Based on the hygiene hypothesis association between atopy and bacillus Calmette Guerin (BCG), purified protein derivative skin test reaction, mycobacterial disease, and environmental mycobacteria are summarized. The role of mycobacterial species in the activation of the innate immune response through Toll-like receptors is mentioned. The implications and perspectives of BCG as a potential therapeutic adjuvant in atopic disease are discussed. PMID- 16701152 TI - Seeing an anesthetic revolution: ocular anesthesia in history. AB - Each surgical procedure places unique demands on the anesthesiologist to create surgical anesthesia with minimal physiologic trespass on the patient as well as the surgical repair. In surgery of the eye, the quest for an anesthetic that does not harm the eye or the patient can be a challenge. The removal of cataracts is one of the most frequently performed operations in the United States, and the majority of patients requiring the procedure are elderly and often have other significant medical conditions. PMID- 16701153 TI - Pharmacology of local anesthetics. AB - Local anesthesia is a common technique used to provide anesthesia for ocular surgical procedures. Despite improvements in operative conditions, there is still the potential for complications, both local and systemic, during routine procedures. This article discusses the physiology of neuronal function, the chemistry of various local anesthetic agents, and the pathogenesis of toxicity. It is vital for the practitioner to have thorough knowledge of these subjects in order to reduce risks when performing ophthalmic procedures. PMID- 16701154 TI - Preoperative medical testing and preparation for ophthalmic surgery. AB - The prevention of complications during and after procedures is the most important goal of preoperative evaluation. Several studies have proven the utility of a patient history and physical examination when making a diagnosis. This article discusses preoperative testing and risk assessment as well as management for various conditions that pose challenges to anesthesia and or surgery. PMID- 16701155 TI - General anesthesia for ophthalmic surgery. AB - Skillful anesthetic management is integral to optimal outcomes after ophthalmic surgery. Although the majority of ophthalmic operations in the United States are performed with local anesthetic techniques, nonetheless general anesthesia may be either necessary or advisable in several challenging circumstances. Ophthalmic patients are often at the extremes of age, and not uncommonly have extensive associated systemic or metabolic diseases. Because the complications of ophthalmic anesthesia can be vision threatening or life threatening, it is imperative that the ophthalmologist and the anesthesiologist understand the complex and dynamic interaction among patient diseases, anesthetic agents, ophthalmic drugs, and surgical manipulation. Effective communication and planning among all involved are essential to safe and efficient perioperative care. PMID- 16701156 TI - Sedation techniques in ophthalmic anesthesia. AB - Sedation/analgesia for ophthalmologic surgery is safe and effective. Ideal sedation can be achieved by careful intravenous titration while monitoring the effect of the appropriate sedative and analgesic agents. The choice of sedation/analgesia strategy should be based on patient preference and the assessment of risk for adverse events. Thorough preoperative screening and preparation of the patient is most important in obtaining cooperation and patient acceptance. PMID- 16701157 TI - Choices of local anesthetics for ocular surgery. AB - This article will help clinicians to be aware of their choices of local anesthetic solutions before they make their ocular anesthetic plan based on the specific requirements of the patient, the surgical procedure, and the properties of the local anesthetic. Choices of local anesthetic solutions and additives for both topical anesthesia and conduction blockade are discussed. PMID- 16701158 TI - Sub-Tenon's Anesthesia. AB - The sub-Tenon's anesthesia block was reintroduced into clinical practice in the early 1990s as a simple, effective, and safe alternative to needle blocks. The technique has remained simple and effective but has evolved. Although still very unusual, both sight- and life-threatening complications have occurred. To safely perform the block, detailed knowledge of anatomy and methods for administering anesthesia are essential. PMID- 16701159 TI - Orbital regional anesthesia. AB - Orbital regional anesthesia is a useful and safe modality for providing excellent operating conditions for the surgeon and painless, pleasant circumstances for the patient. It is especially suited for patients who are extremely sensitive and who could not tolerate topical anesthesia or a sub-Tenon's block without deep sedation. Both intraconal and extraconal techniques can be used safely and effectively if proper precautions are taken to enter the safest areas of the orbit and to avoid the vascular areas and the deep orbit where structures are tightly packed and thus more easily harmed. Thorough knowledge of orbital anatomy and understanding of the globe-orbit relationship of every patient are necessary to perform this form of regional anesthesia. In addition, knowledge of the effects and side effects of the anesthetics and adjuvants is also required. PMID- 16701160 TI - Choosing anesthesia for cataract surgery. AB - Advances in cataract surgery techniques have presented surgeons with new options for ocular anesthesia. As cataract removal has become faster, safer, and less traumatic, the need for akinesia and anesthesia has declined significantly. General anesthesia or retrobulbar block have largely been replaced with other safer and equally effective means of local anesthesia. These newer and less invasive methods have reduced the potential for catastrophic surgical complications, increased the efficiency of cataract surgery, and hastened the process of visual rehabilitation. Today there are numerous modes of anesthesia from which a surgeon can choose. This article reviews the current choices for ocular anesthesia, compares their efficacies, and provides a framework, helping to select the most appropriate type of anesthesia for each patient. PMID- 16701161 TI - Anesthesia considerations for vitreoretinal surgery. AB - The vast majority of vitreoretinal procedures can be safely, comfortably, and efficiently performed under local anesthesia with minimal sedation. Compared with general anesthesia, properly performed monitored local anesthesia offers the patient an increased level of safety, reduced recovery times, and prolonged postoperative pain relief. Nonetheless, the choice of anesthesia technique must be based on the needs of the patient, the requirements of the surgeon, and the skills of the anesthesia provider, ever keeping in mind that our ultimate goal is a satisfied patient with a good visual outcome. PMID- 16701162 TI - Anesthesia for glaucoma surgery. AB - Injections of local anesthetic behind the globe could potentially damage the optic nerve, resulting in visual field defects or even blindness. Glaucoma patients may be at increased risk of this occurring, because of increased susceptibility to pressure/ischemic damage. In extreme cases, this may manifest as visual field "wipe-out." Because of these concerns, "newer" techniques of anterior placement of anesthetic have been promoted, including subconjunctival, anterior sub-Tenon's, topical, and intracameral anesthesia. There remains some controversy regarding the effect of different anesthesia techniques on complication and failure rates for glaucoma surgery. PMID- 16701163 TI - Oculoplastic and orbital surgery. AB - Familiarity with facial anatomy, anesthetic agents, and techniques allows the surgeon to maximize his or her surgical success. Reduction of perioperative complications such as an uncooperative or agitated patient may be reduced with proper anesthesia techniques. This ultimately leads to higher success rates and increased patient and physician satisfaction. PMID- 16701164 TI - Anesthesia for pediatric ocular surgery. AB - Ophthalmic pathology in infants and children undergoing eye surgery ranges from the rare and atypical to the commonplace. These pathologies include nasolacrimal duct obstruction, strabismus, congenital or traumatically induced cataracts, penetrating eye injuries, glaucoma, retinopathy of prematurity, intraorbital tumors, and more. Nasolacrimal duct stenosis, cataracts, and traumatic eye injuries often occur in otherwise healthy pediatric patients; however, many ophthalmopathies can be associated with other congenital disorders that may have important anesthesia implications. In this article, we will review pertinent anesthesia issues within the context of various ophthalmic diseases. PMID- 16701165 TI - Succinylcholine and the open eye. AB - The use of succinylcholine in ocular trauma is controversial because it raises intraocular pressure. This article reviews the advantages and disadvantages of succinylcholine and its alternatives, including regional anesthesia for open globe injuries. Finally, an algorithm is proposed for airway management of patients with penetrating eye injuries, highlighting circumstances where succinylcholine may be the safest muscle relaxant. PMID- 16701166 TI - Management of a blind painful eye. AB - Debilitating ocular pain poses a significant challenge to the ophthalmologist. When the pain is intractable and the eye has very poor vision and is disfigured, surgical removal of the eye has traditionally been the definitive treatment of choice. Because many people are uncomfortable psychologically with removal of their eye, however painful, and other patients are not good surgical candidates, an alternative to enucleation is sometimes warranted, and injection of a neurolytic substance can often induce long-lasting anesthesia for a blind painful eye. This article reviews a range of options for management of blind painful eye from anesthesia to enucleation. PMID- 16701167 TI - Complications of anesthesia for ocular surgery. AB - This article discusses the two types of anesthesia used during ocular surgery and their associated complications. PMID- 16701168 TI - Economic evaluation of different systems for cataract surgery and anesthesia. AB - Economic evaluation is an increasingly important component of health and medical care policy making although it continues to be met with some resistance. Many fields of medical care services and public health have extensive economic evaluation literatures. In ophthalmology, the literature is less well developed and there is an ongoing discussion of the most appropriate methods. This article outlines different types of economic evaluations providing examples on their potential use in ophthalmic care decision making, reviews three articles in the brief recent literature on the cost-effectiveness of ophthalmic anesthesia and cataract surgery in the United States with a focus on explaining methods that were used, and discusses ways in which research in this area might be moved forward. PMID- 16701169 TI - Application of stem cells for cardiovascular grafts tissue engineering. AB - Congenital and acquired heart diseases are leading causes of morbidity and mortality world-wide. Currently, the synthetic materials or bioprosthetic replacement devices for cardiovascular surgery are imperfect and subject patients to one or more ongoing risks including thrombosis, limited durability and need for reoperations due to lack of growth in children and young adults. Suitable replacement grafts should have appropriate characteristics, including resistance to infection, low immunogenicity, good biocompatability and thromboresistance, with appropriate mechanical and physiological properties. Tissue engineering is a new scientific field aiming at fabrication of living, autologous grafts having structure or function properties that can be used to restore, maintain or improve tissue function. The use of autologous stem cells in cardiovascular tissue engineering is quite promising due to their capacity of self-renewal, high proliferation, and differentiation into specialized progeny. Progress has been made in engineering the various components of the cardiovascular system, including myocardial constructs, heart valves, and vascular patches or conduits with autologous stem cells. This paper will review the current achievements in stem cell-based cardiovascular grafts tissue engineering, with an emphasis on its clinical or possible clinical use in cardiovascular surgery. PMID- 16701170 TI - Murine dendritic cells that are resistant to maturation are unable to induce tolerance to allogeneic stem cells. AB - Induction of donor-specific hyporesponsiveness would minimize the need for intensive immunosuppression in the clinical setting of graft rejection and dendritic cells (DCs) might be useful tools for this purpose. Besides their ability to induce immunogenic T-cell responses, these antigen presenting cells can lead to T-cell anergy, when antigen presentation occurs in the absence of costimulation as is the case in immature DCs (iDCs). In continuance of publications reporting on the use of iDCs to induce tolerance to various organs, we set out to determine whether tolerance could be induced in a model of allogeneic stem cell transplantation. Immature DCs were obtained by culture with very low concentrations of GM-CSF and by treating DCs with Dexamethasone (Dex). We show that these DCs express low levels of MHCII and costimulatory molecules and that this immature phenotype is retained after application of maturation stimuli. We also prove that these alternatively activated DCs are unable to induce T-cell proliferation in vitro. When used in vivo however, these tolerogenic DCs do not provide tolerance to fully mismatched or haploidentical stem cells. PMID- 16701172 TI - Application of popliteal lymph node assay to evaluate tolerogenic effect of donor leukocyte infusion. AB - BACKGROUND: Popliteal lymph node assay (PLNA) has long been proposed to detect immunostimulating potential of chemicals. Here, the PLNA was used to evaluate the effect of donor leukocyte infusion on recipients' reaction to donor-specific antigens. METHODS: Donor rats' peripheral blood leukocytes (ranging from 1 x 10(4) to 500 x 10(4) cells) were intravenously (i.v.) infused into recipients. A week later recipients' reaction to donor-specific antigen was evaluated, using the PLNA technique, by subcutaneous injection of donor spleen cells to one hind footpad of recipients and injection of saline to the other. Seven days later all recipients were killed and their PLNs' weight and cellularity indices were determined. While the same process was applied to the positive control (PC) animals, rats without leukocyte infusion, negative control (NC) animals, rats without leukocyte infusion, were injected in both hind footpads with saline. RESULTS: The PLN weight indices of recipients of: > or =5 x 10(4) leukocytes were significantly lower than PC animals (P < 0.001), whereas the weight indices of recipients of 1 x 10(4) cells were similar to PC group but higher than NC animals (P < 0.0001). However, the PLN cellularity indices of recipients of < or =10 x 10(4) cells were not different from PC animals but the PLN cellularity indices of recipients of: > or =50 x 10(4) cells were significantly lower than PC group (P < 0.05). CONCLUSION: Overall, these results suggest that donor leukocytes infusion dose-dependently decrease reaction to donor-specific antigens, but a state of tolerance to donor antigen might be induced at the dose of: > or =50 x 10(4) cells. PLNA appears to represent a simple test model to quantify efficacy of immunotolerance protocols. PMID- 16701171 TI - Low molecular weight fucan prevents transplant coronaropathy in rat cardiac allograft model. AB - INTRODUCTION: Transplant arteriosclerosis is the main cause of long-term failure after cardiac transplantation. Vascular rejection is thought to be due to intimal proliferation occurring in response to arterial wall immune-mediated injury. A low molecular weight fucan (LMWF) compound, a sulfated polysaccharide, has been demonstrated to increase plasma levels of stromal cell-derived factor 1 (SDF-1) and consequently to mobilize bone marrow-derived vascular progenitor cells (BMVPC). The aim of this study was to evaluate the capacity of LMWF to prevent coronary intimal proliferation in a rat cardiac allograft model. METHODS: Heterotopic abdominal cardiac graftings were performed in Brown Norway (BN) and Lewis (LEW) rats. Animals were divided into 4 groups of 10 rats. Two groups were treated intramuscularly with LMWF (5 mg/kg/day) (one BN to BN isograft group, and one BN to LEW allograft group); and two control groups were LMWF-untreated (one BN to BN isograft group and one BN to LEW allograft group). All animals were treated by cyclosporin (15 mg/kg/day) sub-cutaneously and sacrificed at day 30. The cardiac grafts were assessed by morphometry of structural parameters and by histological and immunohistochemical analyses. RESULTS: All cardiac isografts were devoid of any coronary and parenchymal lesions. In contrast, the majority of untreated allografts developed coronary intimal proliferation in close association with intimal and adventitial inflammatory CD68(+) cell infiltration. Further, the parenchyma exhibited large areas of actin(+) cells (myofibroblasts) of recipient origin colocalized with the CD68(+) infiltrating cells. Interestingly, all LMWF-treated allografts were well protected against coronary and parenchymal lesions and the coronary arteries exhibited an intimal monolayer of flat cells, which however were CD34 negative. CONCLUSION: treatment with LMWF appeared very effective in this rat cardiac allograft model to prevent arterial and parenchymal lesions occurring in response to alloimmune injury. However this protective effect does not appear to depend on mobilization of bone marrow derived cells. PMID- 16701173 TI - Toll-like receptors and graft rejection. AB - Innate immunity plays a role in fighting against invading microorganisms. Emerging evidence suggests that in addition to responding to pathogen-associated molecular patterns of microorganisms, Toll-like receptors (TLRs) can be activated by endogenous ligands expressed by mammalian cells. Clinical and laboratory studies have shown that TLRs may participate in organ graft rejection and transplant immune tolerance, which are briefly reviewed in the present manuscript. PMID- 16701175 TI - Prediction of acute renal allograft rejection in early post-transplantation period by soluble CD30. AB - To evaluate the feasibility of serum sCD30 for prediction of acute graft rejection, we analyzed clinical data of 231 patients, whose serum levels of sCD30 were detected by ELISA before and after transplantation. They were divided into three groups: acute rejection group (AR, n = 49), uncomplicated course group (UC, n = 171) and delayed graft function group (DGF, n = 11). Preoperative sCD30 levels of three groups were 183 +/- 74, 177 +/- 82 and 168 +/- 53 U/ml, respectively (P = 0.82). Significant decrease of sCD30 was detected in three groups on day 5 and 10 post-transplantation respectively (52 +/- 30 and 9 +/- 5 U/ml respectively, P < 0.001). Compared with Group UC and DGF, patients of Group AR had higher sCD30 values on day 5 post-transplantation (92 +/- 27 U/ml vs. 41 +/- 20 U/ml and 48 +/- 18 U/ml, P < 0.001). However, sCD30 levels on day 10 post transplantation were virtually similar in patients of three groups (P = 0.43). Receiver operating characteristic (ROC) curve demonstrated that sCD30 level on day 5 post-transplantation could differentiate patients who subsequently suffered acute allograft rejection from others (area under ROC curve 0.95). According to ROC curve, 65 U/ml may be the optimal operational cut-off level to predict impending graft rejection (specificity 91.8%, sensitivity 87.1%). Measurement of soluble CD30 on day 5 post-transplantation might offer a noninvasive means to recognize patients at risk of impending acute graft rejection during early post transplantation period. PMID- 16701174 TI - CD40L, CD28, and CTLA-4 expression on CD4+ T cells in kidney graft recipients: a relationship with post-transplantation clinical course. AB - BACKGROUND: Experimental studies have demonstrated that the intensity of alloreactivity against a transplanted organ results from an interaction of positive (CD40/CD40L and B.7/CD28) and inhibitory (B.7/CTLA-4) signals between antigen-presenting cells (APCs) and T lymphocytes. METHODS: We examined the CD40L, CD28, and both surface (s) and intracellular (i) CTLA-4 expressions on freshly drawn and anti-CD3+rIL-2-stimulated peripheral blood CD4+ T cells in groups of kidney transplant recipients in relation to distinct clinical course using the tri-color immunofluorescence method. RESULTS: The median proportions of freshly isolated CD3+/CD4+/CTLA-4+ and CD3+/CD4+/CD40L+ cells in all groups of graft recipients were higher than in control subjects. In patients with stable graft function (SGF), non-significantly higher sCTLA-4, significantly higher iCTLA-4 expression, and significantly lower CD40L expression on freshly drawn CD4+ T cells compared with recipients with chronic allograft nephropathy (CAN) were found. Moreover, CD4+ T cells from SGF patients showed a higher potential to express sCTLA-4 and CD40L molecules and to down-regulate the CD28 molecule in response to ex vivo stimulation than those from patients with CAN. In patients without acute graft rejection (NAGR), a markedly higher proportion of freshly drawn CD3+/CD4+/iCTLA-4+ cells compared with patients with acute graft rejection (AGR) and an up-regulation of the median percentage of CD3+/CD4+/CD40L+ cells after ex vivo stimulation was found. CONCLUSIONS: In patients with SGF, peripheral blood CD4+ T cells exhibited a higher potential to express surface CTLA-4 and CD40L and to down-regulate CD28 costimulatory molecules in response to ex vivo stimulation, indicating a relationship between the expression patterns of both costimulatory and inhibitory molecules in CD4+ T cells and clinical course after renal transplantation. PMID- 16701176 TI - Everolimus and reduced cyclosporine trough levels in maintenance heart transplant recipients. AB - BACKGROUND: Long-term survival of patients after oHTX significantly increased over the last years, but CAV and chronic renal failure due to nephrotoxic side effects of CNIs still remain unsolved problems. Everolimus has shown to reduce acute cellular rejection and may allow CsA dosage reduction. In this study the effectiveness of Everolimus in combination with CsA dosage reduction in maintenance oHTX immunosuppression and the influence on renal function was tested. METHODS: 37 patients (30 male, 7 female) after oHTX were divided into group A (n = 20) receiving Everolimus in combination with CsA and prednisolone and group B (n = 17) under standard immunosuppression with CsA, MMF and prednisolone. Patients received 1.0 mg to 1.5 mg Everolimus per day and target Everolimus trough levels were between 3 and 8 ng/ml. Death, safety, side effects, BPAR, trough levels, and routine laboratory values especially creatinine levels were monitored over a follow-up period of 8 months retrospectively and statistically evaluated. RESULTS: A significant reduction of CsA dosage (p < 0.001) and a significant CsA trough level reduction (p < 0.001) to a median CsA trough level of 68.5 ng/ml were achieved in group A. Mean Everolimus trough levels were reached within 1 week and 2 months. Renal function was stable in both groups. No statistical differences in BPAR, hospitalization rates or triglyceride levels were observed. Cholesterol levels significantly increased in group B (p = 0.024). CONCLUSION: CsA trough levels and dosage can be significantly reduced in combination with Everolimus without higher rejection rates and with stable kidney function in oHTX patients. PMID- 16701177 TI - Clinical factors influencing T-cell receptor excision circle (TRECs) counts following allogeneic stem cell transplantation in adults. AB - To ascertain the clinical factors involved in T-cell reconstitution after allogeneic stem cell transplantation (SCT), we evaluated serial assessments of lymphocyte subsets by flow cytometry and TRECs levels by quantitative PCR in 83 adult patients. Patient age >25 years, unrelated donor, CMV infection and acute graft-versus-host disease (GVHD) adversely affected CD3(+) and CD8(+) T-cell recovery after SCT (p < 0.05). TRECs were low or undetectable during the first months after transplant and progressively increased thereafter. However, median TRECs of patients did never achieve normal values compared to healthy donors (median follow-up 9 months, range 2-42). Presence and severity of chronic GVHD significantly affected TRECs counts: patients with chronic GVHD had lower TRECs than patients without GVHD at 9, 12 and 24 months after SCT (p = 0.002, p = 0.022, p = 0.015). Patients with limited chronic GVHD had higher TRECs compared to patients with extensive GVHD (p = 0.018). No relationship was observed between fungal or bacterial infections and TRECs. Nonetheless, CMV infection was associated with lower TRECs (p = 0.032). Our data support the concept that adult thymus contributes with a slow but continuous production of thymic T cells to immune reconstitution after SCT. Chronic GVHD is the main factor associated to a delay in TRECs counts recovery. PMID- 16701178 TI - Correlation between acute rejection severity and CD8-positive T cells in living related liver transplantation. AB - The Banff schema is the most widely used standard grading system for liver allograft rejection. To investigate the relationship between the Banff rejection activity index (RAI) and the presence of lymphocyte subpopulations in allograft liver tissue, assuming these cells to probably play an important role in the mechanism of acute cellular rejection (ACR), we performed immunohistological examinations using liver tissues with various ACR severities after living related liver transplantation (LRLT). In total, 37 needle liver biopsy specimens with ACR in LRLT were examined using antibodies to CD4, CD8, and CD20. Formalin-fixed and paraffin-embedded liver tissues were used to maintain morphology. Immunohistological findings and RAI score according to the Banff schema were compared. In the results, mainly CD8-positive (CD8+), rather than CD4-positive (CD4+), cells were detected in the portal tract and were also found in bile duct epithelium and subendothelial areas of portal veins. The number of CD8+ cells increased according to ACR grade, whereas CD4+ cells tended to decrease. There were significant correlations between the presence of CD8+ cells (p = 0.0006) and CD4+ cells (p = 0.0003) and ACR severity. On the other hand, CD20-positive cells did not correlate with ACR severity (p = 0.472). The results indicate that CD8+ cells play important roles in ACR severity of LRLT, suggesting the number of CD8+ cells in liver tissue to be useful as a supplementary tool, in addition to RAI of the Banff schema, for objective evaluation of ACR severity. PMID- 16701179 TI - The complete mitochondrial genome of an agamid lizard from the Afro-Asian subfamily agaminae and the phylogenetic position of Bufoniceps and Xenagama. PMID- 16701180 TI - Sleeping beauty or big bad wolf? PMID- 16701181 TI - Multiple injections in axillary block: where and how many? PMID- 16701182 TI - Thoracic paravertebral block: influence of the number of injections. AB - BACKGROUND AND OBJECTIVES: The purpose of this study was to assess the radiographic and clinical distribution of 1 to 4 paravertebral injections by use of the same total volume of local anesthetic mixture. METHODS: Sixty-nine patients scheduled for surgical interventions suitable for bilateral PVB were included in the study and were randomly assigned to 1 of 3 treatment groups. Group 1 received 4 paravertebral injections on one side versus 1 injection on the contralateral side (23 patients), group 2 received 4 injections versus 2 injections (23 patients), and group 3 received 4 injections versus 3 injections (23 patients). RESULTS: Sixty-one patients were included in the final analysis, which resulted in 368 thoracic paravertebral injections. Overall, 97% of the patients had adequate loss of sensation within the targeted area at the side of 4 injections, compared with only 11% for the single injections. The average vertical spread of contrast was found to be significantly greater in the set of 4 injections, with mean (SD) 6.5 (2.01) dermatomes, compared with the single injection, with 3.0 (1.19) dermatomes (P < .05). The average vertical spread of contrast and complete absence of sensation were significantly greater in the set of 4 injections compared with the set of 2 and 3 injections (P < .05). CONCLUSION: The main finding of the present study was that multiple paravertebral injections resulted in more reliable radiographic and clinical distribution compared with a single-injection technique. PMID- 16701183 TI - A comparison of stimulation patterns in axillary block: part 2. AB - BACKGROUND AND OBJECTIVES: Radial plus musculocutaneous nerve stimulation may have a predominant role in the success of an axillary block, producing more extensive anesthesia of the upper limb than median plus musculocutaneous nerve stimulation. However, no comparison has been made with ulnar plus musculocutaneous nerve stimulation. We compared the extent of both sensory and motor block after ulnar plus musculocutaneous nerve stimulation or radial plus musculocutaneous nerve stimulation. METHODS: Sixty patients were randomly assigned to receive an axillary block using either radial plus musculocutaneous or ulnar plus musculocutaneous nerve stimulation with 40 mL plain 1.5% mepivacaine. Patients were assessed for sensory block by the pinprick method at 5 and 20 minutes. RESULTS: No statistically significant differences were found in the rates of anesthesia at 20 minutes in the cutaneous nerve distributions of the upper limb between radial plus musculocutaneous and ulnar plus musculocutaneous nerve stimulation except for the following nerves: radial (90% and 63.3%, respectively), medial cutaneous of the forearm (83.3% and 100%, respectively), and medial cutaneous of the arm (73.3% and 93.3%, respectively). Global sensory score (minimum: 0; maximum: 12 points) at 20 minutes was significantly higher after radial plus musculocutaneous than after ulnar plus musculocutaneous nerve stimulation: 12 (11-13) and 11 (10-12), respectively. The rates of median nerve blockade were 50% and 53%, respectively. CONCLUSIONS: Radial plus musculocutaneous nerve stimulation produced more extensive anesthesia of the upper limb than did ulnar plus musculocutaneous nerve stimulation. However, there is not an optimal combination of 2 responses in axillary brachial plexus block. PMID- 16701184 TI - A comparison of mepivacaine versus lidocaine for episcleral (sub-tenon's) block for cataract surgery in an ambulatory setting. AB - BACKGROUND AND OBJECTIVES: For eye surgery, motor block is still often requested by the surgeon. For cataract surgery, rapid block resolution allows eyelids to move and allows eye-patch removal. Therefore, short-duration block is useful in early rehabilitation for ambulatory surgery. Lidocaine is classically assumed to have shorter duration than mepivacaine. Therefore, lidocaine alone might be considered as an alternative to mepivacaine. METHODS: In this randomized, double blind study, we compared mepivacaine 2% (n = 22) and lidocaine 2% (n = 25) in 47 patients who received episcleral (sub-Tenon's) block for cataract surgery. Akinesia score was measured 1, 5, 10, and 15 minutes and 1, 2, 4, and 6 hours after the end of injection. Primary outcome was block duration (time from injection to full recovery). Secondary outcomes were time to block onset and best akinesia score for each patient. Complications were recorded. RESULTS: The 2 groups were similar for demographic and anesthetic features. We observed no significant difference between mepivacaine and lidocaine in terms of onset, quality of akinesia, and block duration. One case of ocular hypertonia and 1 case of strabismus were observed in the lidocaine group, which could be imputed to hyaluronidase unavailability during the study period or to increased lidocaine myotoxicity. CONCLUSIONS: We found no argument to favor lidocaine over mepivacaine in episcleral (sub-Tenon's) eye block, especially in terms of motor block duration. PMID- 16701185 TI - Infraorbital nerve block in children: a computerized tomographic measurement of the location of the infraorbital foramen. AB - BACKGROUND AND OBJECTIVES: Infraorbital nerve blocks are performed in children undergoing cleft-lip surgery and endoscopic sinus surgery. The nerve exits the maxilla at the level of the infraorbital foramen. A sensory block of the infraorbital nerve can be performed by use of an intraoral route or an extraoral route. The objective of this study was to derive a mathematical formula to determine the position of the infraorbital foramen. METHODS: Computerized tomographic (CT) scans of children who were admitted to our institution for various reasons were evaluated with the assistance of a pediatric neuroradiologist. The distance of the foramen from the midline was determined, and a mixed-effects linear-regression model was used (PROC MIXED in SAS 9.1) to ascertain whether the distance of the infraorbital foramen correlated linearly with the age of the patient. RESULTS: CT scans of 48 pediatric patients were evaluated. The age of the patient accounted for more than half of the variation in this model. Distance of the infraorbital foramen can be predicted by use of the following formula: [Distance of the infraorbital foramen (in mm) from midline = 21.3 + 0.5 x age (in years)]. CONCLUSIONS: Our study found a linear correlation with age to the distance of the infraorbital foramen. This anatomic knowledge of the location of the infraorbital foramen may facilitate correct positioning of the needle. Future studies on the use of this formula with anatomic coordinates should be performed to test the validity of this formula. PMID- 16701186 TI - A subgluteal approach to the sciatic nerve in adults at 10 cm from the midline. AB - BACKGROUND AND OBJECTIVES: In 2003 we introduced the concept of a sciatic nerve block performed in the midgluteal area at a fixed distance from the midline in all adults regardless of gender and/or body size. The anatomic basis for that study suggested that a subgluteal block could also be accomplished in a similar fashion. METHODS: After informed consent, 20 patients were prospectively recruited. Patients were positioned in lateral decubitus. The needle insertion site was located in the subgluteal fold at 10 cm from the midline. The needle was advanced parallel to the midline until a sciatic nerve response was elicited. With a visible response at 0.5 mA, 30 mL 1.5% mepivacaine plus 1:200,000 epinephrine was slowly injected. Sensory anesthesia was tested on the plantar and dorsal aspects of the foot as well as the posterior thigh. RESULTS: Residents performed all blocks. The approach was 100% successful in locating the sciatic nerve with 3 attempts or less from a site located 10 cm from the midline. The block provided successful surgical anesthesia in 90% of the cases; 2 cases required local anesthetic supplementation. Only 3 patients developed anesthesia of the posterior thigh within 30 minutes of injection. CONCLUSIONS: This report shows that a sciatic nerve block can be performed in the subgluteal area at 10 cm from the midline in adult patients of both sexes and various sizes. Anesthesia of the posterior thigh is not consistently accomplished with this approach. PMID- 16701187 TI - Analgesic efficacy of two doses of intrathecal midazolam with bupivacaine in patients undergoing cesarean delivery. AB - OBJECTIVES: In this prospective, randomized, double-blind, placebo-controlled study, we investigated the postoperative analgesic efficacy of 2 doses of intrathecal midazolam as an adjunct to bupivacaine for spinal anesthesia. METHODS: Sixty patients undergoing elective cesarean delivery under spinal anesthesia were allocated randomly to 3 groups: group B, 2 mL hyperbaric bupivicaine 0.5%; group BM1, 2 mL bupivacaine plus midazolam 1 mg (preservative free); and group BM2, 2 mL bupivicaine plus midazolam 2 mg. RESULTS: The mean duration of postoperative analgesia (determined by request for rescue medication) was 3.8 +/- 0.5 hours in group B compared with 4.3 +/- 0.7 hours in group BM1 (P = .18), and 6.1 +/- 1.0 hours in group BM2 (P = .001). Supplemental analgesic requirements with diclofenac were significantly less in group BM2 (93 +/- 29 mg) compared with group B (145 +/- 12 mg) and group BM1 (148 +/- 16 mg, P < .001). Time to block regression was longer in group B (182 +/- 30 minutes) compared with group BM1 (152 +/- 32 minutes) and group B (126 +/- 20 minutes) (both P < .001). Arterial pressure, heart rate, oxygen saturation, sedation score, and time to first void were comparable between groups. Group B had a significantly higher incidence of nausea and vomiting than groups BM1 and BM2 (P = .02). No neurologic deficits were observed. CONCLUSIONS: Intrathecal midazolam 2 mg provided a moderate prolongation of postoperative analgesia when used as an adjunct to bupivacaine in patients undergoing cesarean delivery. Intrathecal midazolam, 1 mg and 2 mg, decreased postoperative nausea and vomiting. PMID- 16701188 TI - Preemptive analgesia with bupivacaine for segmental mastectomy. AB - BACKGROUND AND OBJECTIVES: Preemptive analgesia is the concept of providing analgesia before surgical incision, resulting in less postoperative pain. The purpose of this study is to determine if preemptive and/or postoperative local anesthetic infiltration of bupivacaine in patients undergoing segmental mastectomy results in less postoperative pain compared with patients receiving placebo. METHODS: In this prospective, double-blinded study, 120 patients were randomized into 4 groups: group 1, preincisional (10 mL) and postoperative (10 mL) wound infiltration of 0.5% bupivicaine, (+Pre+Post); group 2, preincisional bupivacaine (10 mL) and postoperative infiltration (10 mL) of placebo (normal saline solution), (+Pre-Post); group 3, preincisional placebo (10 mL) and postoperative bupivacaine (10 mL), (-Pre+Post); or group 4, preincisional (10 mL) and postoperative infiltration of placebo (10 mL), (-Pre-Post). All patients received a standardized laryngeal mask general anesthetic. Data were recorded at the following time intervals: preoperative admission, postanesthesia care unit (PACU) admission, PACU stay, stepdown-unit admission, stepdown-unit stay, hospital discharge, and 24 hours post operation. RESULTS: No difference was noted with respect to preoperative pain visual analog scale (VAS, 0-100 mm), surgical duration, PACU stay time, stepdown-unit stay time, incidence of postoperative nausea, or treatment for nausea in all measured time periods. The placebo group (group 4) had significantly higher mean pain VAS scores during the early postoperative period (PACU admission and PACU stay) compared to the other groups (PACU admission: group 1 = 2 +/- 8, group 2 = 4 +/- 11, group 3 = 3 +/- 15, group 4 = 17 +/- 21, P < .01; PACU stay: group 1 = 6 +/- 13, group 2 = 6 +/- 10, group 3 = 10 +/- 21, group 4 = 20 +/- 18, P < .01). Likewise, the number of patients who reported pain (pain frequency) was significantly higher in group 4 (placebo) compared with all other groups at PACU admission, PACU stay, stepdown-unit admission, and stepdown-unit stay (P < or = .01). CONCLUSION: Preincisional and/or postoperative wound bupivacaine infiltration lacks preemptive analgesic effects for segmental mastectomy. PMID- 16701189 TI - Magnesium added to prilocaine prolongs the duration of axillary plexus block. AB - BACKGROUND AND OBJECTIVE: The aim of this study was to evaluate the effect of the addition of magnesium to prilocaine on the duration of sensory and motor-nerve block. METHODS: After institutional approval, 60 ASA physical status I and II patients, between 18 and 60 years of age and scheduled for forearm and hand surgery under axillary brachial plexus block, were included in the study. Patients were randomly assigned to 1 of 4 groups. All of the patients received 5 mg/kg of 2% prilocaine and isotonic sodium chloride solution in 35 mL total volume for axillary brachial plexus block. Group I received intravenous saline, and group II received 150 mg intravenous magnesium at the same time as local anesthetic administration. In group III, 100 mg of magnesium, and in group IV, 150 mg of magnesium, were added to local anesthetic solution. Sensory block and motor block of musculocutaneous, radial, median, and ulnar nerves were recorded at 5-minute intervals. RESULTS: The duration of motor block was significantly longer in group IV than in other groups (167 +/- 30, 177 +/- 17, 180 +/- 20, and 250 +/- 19 minutes in groups I to IV, respectively) (P < .01). Mean duration of sensory block in both of the perineural magnesium groups was statistically different than in groups I and II (P < .001). Duration of sensory block in group IV (304 +/- 30 minutes) was significantly longer than in group III (253 +/- 23 minutes) (P < .001). CONCLUSION: The admixture of magnesium to prilocaine for axillary brachial plexus block provided a pronounced prolongation of sensory and motor block without side effects. PMID- 16701190 TI - The analgesic effects of perioperative gabapentin on postoperative pain: a meta analysis. AB - BACKGROUND AND OBJECTIVES: Gabapentin is an anticonvulsant that has been shown to be effective in the treatment of neuropathic and inflammatory pain in animal and human studies. The analgesic effect of its perioperative use has not been fully elucidated. METHODS: This systematic review (meta-analysis) included 12 randomized controlled trials of 896 patients undergoing a variety of surgical procedures that investigated the impact of perioperative administration of gabapentin on postoperative outcome. RESULTS: The pooled visual analog scores for pain at 4 hours and 24 hours were significantly less in those patients who received gabapentin (weighted mean difference [WMD] = -1.57; 95% confidence interval [CI], -2.14 to -0.99 and WMD = -0.74; CI, -1.03 to -0.45, respectively). A concomitant decrease in opioid usage by those patients who received gabapentin was also noted (odds ratio [OR] = -17.84; CI, -23.50 to -12.18). Gabapentin administration was associated with sedation and anxiolysis (OR = 3.28; CI, 1.21 8.87) but not associated with a difference in lightheadedness, dizziness, nausea, or vomiting. CONCLUSIONS: Based on this systematic review, perioperative oral gabapentin is a useful adjunct for the management of postoperative pain that provides analgesia through a different mechanism than opioids and other analgesic agents and would make a reasonable addition to a multimodal analgesic treatment plan. PMID- 16701191 TI - Which spinal cutaneous nociceptive neurons are inhibited by intravenous lidocaine in the rat? AB - BACKGROUND AND AIMS: Intravenous lidocaine (IVL) produces analgesia in multiple painful disorders. The neurophysiological effects of IVL are not well defined, but studies in visceral nociceptive systems have shown that IVL has differential effects on subgroups of spinal neurons. The present study determined whether a similar differential effect of IVL occurs in spinal neurons excited by noxious cutaneous stimuli. METHODS: In decerebrate, cervical spinal cord-transected rats, the lumbosacral spinal cord was exposed by a laminectomy. Single-unit recordings were made of dorsal horn neurons excited by noxious cutaneous stimuli. Each neuron's response to noxious (pinch) and nonnoxious (brush) cutaneous stimuli were determined and the effect of a counterirritation stimulus (noxious skin pinch presented in the upper body) on spontaneous activity quantified. In a subset of neurons, sequential doses of IVL were administered, and responses of each neuron to repeated 50 degrees C heating of the hindpaw/tail were determined. RESULTS: IVL dose-dependently inhibited neurons excited by heating of the hindpaw/tail. IVL produced significantly greater inhibition of both spontaneous and heat-evoked activity in neurons that did not show counterirritation effects when compared with those neurons that did show counterirritation effects. Standard classification of neurons as wide-dynamic range or nociceptive-specific was less predictive of the IVL effect. CONCLUSIONS: IVL had differential inhibitory effects on 2 spinal cutaneous nociceptive neuron populations. Other drugs could also have differential effects on sensory pathways, suggesting a mechanism whereby certain drugs differentially affect different types of pain. PMID- 16701192 TI - Reduction of pulmonary edema by short-acting local anesthetics. AB - BACKGROUND AND OBJECTIVES: Local anesthetics (LAs) possess a variety of effects that cannot be explained by the typical block of neuronal sodium channels. Antithrombotic effects of LAs are well known, but LAs also act as bactericides. Therefore, an investigation of the influence of LAs on the inflammatory response of the isolated rat lung (n = 78) to an N-formyl-l-leucin-methionyl-l phenylalanine (FMLP) stimulus was performed. METHODS: The experiments were performed on isolated and ventilated rat lungs perfused with cell-free and plasma free buffer. LAs (lidocaine and mepivacaine) were injected in various concentrations before application and activation of human granulocytes by FMLP. Pulmonary arterial pressure (PAP) and lung weight gain were monitored continuously. LAs in final dosages from 10(-2) to 10(-7) mg/kg body weight (n = 6 each) were injected into the pulmonary artery before treatment with FMLP (10(-6) M) to induce pulmonary arterial hypertension. Perfusate samples were taken intermittently to determine thromboxane A(2) (TX A(2)) and endothelin-1 concentrations. Microscopic analyses were performed to assess the degree of lung injury. RESULTS: Pretreatment with LAs significantly reduced the FMLP-induced PAP increase (treatment group v sham group: 0.5 to 5 mm Hg v 8 mm Hg; P < .05) and the release of endothelin-1 (2.4 v 5 fmol/mL). Histologic damage seen as acute granulocytic alveolitis was reduced by lidocaine and mepivacaine, even below clinically relevant concentrations. CONCLUSION: LA pretreatment reduces inflammatory reactions after FMLP stimulus. PMID- 16701194 TI - Cervical transforaminal injection and the radicular artery: variation in anatomical location within the cervical intervertebral foramina. AB - BACKGROUND: Recent articles have detailed the adverse events associated with transforaminal steroid injections into the radicular arteries. Guidelines on strict transforaminal epidural techniques have been described to limit complications. There remains limited information regarding the cervical level of entry, location within the intervertebral foramina, and prevalence of the radicular arteries within the population. METHODS: With the aid of premortem angiography and postmortem latex-injected vasculature, a single detailed cadaveric dissection was performed to elucidate the specific anatomic location of the radicular arteries within the intervertebral foramina and the anastomoses of the arteries to the anterior spinal artery. RESULTS: Five radicular arteries (C5, C6, two at C7, C8) were traced entering the foramina either anteriorly or posteriorly to supply the anterior and posterior spinal arteries. Radicular arteries received blood supply from either the deep cervical (C8) or vertebral arteries. The C8 radicular artery entered the lateral aspect of the foramen and penetrated the dural sleeve within the inferior portion of the foramen, directly inferior to the exiting spinal nerve, to supply the anterior spinal artery. This artery was of a large enough caliber to be entered by a 22-gauge needle. CONCLUSIONS: A larger population is necessary to characterize the range of anatomic variations in arterial supply within the foramina. Available studies support the current technique of fluoroscopic needle insertion. Yet, there is wide anatomic variation in the origin and location of these vessels, and even with strict adherence to technique, it is feasible that a properly placed needle could penetrate a radicular artery. PMID- 16701193 TI - Comparison of analgesic methods for total knee arthroplasty: metabolic effect of exogenous glucose. AB - BACKGROUND AND OBJECTIVES: This study was set up to assess whether postoperative suppression of gluconeogenesis by dextrose infusion would be influenced by continuous peripheral nerve block (CPNB) with local anesthetics, in comparison with epidural and with intravenous analgesia. METHODS: Twenty-seven patients, undergoing elective primary total knee arthroplasty for osteoarthritis, were randomly allocated to one of the three groups of 9 patients each: patient controlled analgesia (PCA) with i.v. morphine, epidural with bupivacaine 0.1% and fentanyl 3mug/ml, or continuous femoral and sciatic blocks with ropivacaine 0.2%. Endogenous glucose production, an index of gluconeogenesis, and glucose clearance, an index of whole body glucose uptake, were assessed on the second postoperative day by measuring [6,6-(2)H(2)]glucose kinetics after an overnight fast (fasted state), and during a 3-h period infusion of dextrose at 4 mg/kg/min (fed state). Visual analog scale (VAS) at rest and at knee flexion, use of morphine, mobilization, nutritional intake, and bowel function were also collected. RESULTS: Endogenous glucose production was totally suppressed by 3 h of dextrose infusion in all 3 groups (P < .001) while glucose clearance was unchanged. Blood glucose and insulin increased (P < .001), while glucagon decreased, with the greatest change in the epidural group (P < .05). VAS at rest and at knee flexion was significantly lower in patients receiving epidural and CPNB compared to i.v. morphine (P < .05). Restoration of bowel function, assessed as return of bowel movements, was faster in the CPNB group (P < .05). CONCLUSION: Excellent analgesia was achieved in the epidural and continuous nerve block groups. Postoperative gluconeogenesis was totally suppressed by dextrose infusion independent of the analgesia technique with no change in glucose utilization. PMID- 16701195 TI - Delayed convulsions and brief contralateral hemiparesis after retrobulbar block. AB - OBJECTIVE: This case report describes convulsions and hemiparesis after retrobulbar injection with good outcome in a patient undergoing outpatient cryocoagulation of his right eye. CASE REPORT: We report a young man in which localized convulsions of the ipsilateral face occurred 9 minutes after retrobulbar injection followed shortly by convulsions of the contralateral arm and leg. After the convulsions, the patient experienced left-sided hemiparesis resolving approximately 1 hour after the injection. There was no hemodynamic instability during this period. It was difficult to determine the exact cause of convulsions and hemiparesis. CONCLUSIONS: We believe these complications occurred because of unintentional injection of local anesthetic agent into the subarachnoid space without affecting the brainstem. Possible mechanisms of spread of local anesthetic agent into the central nervous system after retrobulbar block are discussed. PMID- 16701197 TI - Cerebellar herniation after cervical transforaminal epidural injection. AB - OBJECTIVE: The purpose of this study is to inform readers of potential catastrophic complications associated with performing cervical transforaminal epidural steroid injections. CASE REPORT: A 31-year-old man presented to the pain clinic with a history of cervical radicular pain and right upper-extremity radicular symptoms. He was referred from the orthopedic spine clinic for evaluation for epidural steroid injection, having failed conservative treatment consisting of nonsteroidal anti-inflammatory drugs, muscle relaxants, oral steroids, and physical therapy. After undergoing a transforaminal epidural steroid injection at the right C8 nerve root, he developed a cerebellar infarct and brainstem herniation. He survived but has residual deficits of persistent diplopia on right lateral gaze and difficulties with short-term memory loss and concentration. CONCLUSIONS: Although transforaminal epidural steroid injections are an efficacious treatment for radicular syndromes, there can be catastrophic complications. In light of the growing body of similar case reports, further investigation is warranted to establish a safe protocol for the use of this modality. PMID- 16701198 TI - It's self-regulation, but not as we knew it! PMID- 16701196 TI - Epidural spread after continuous cervical paravertebral block: a case report. AB - BACKGROUND AND OBJECTIVE: This report illustrates epidural spread after continuous cervical paravertebral block (CCPVB). By fluoroscopy, it also explains the mechanism of the complication. CASE REPORT: A healthy 22-year-old male developed bilateral upper-extremity motor weakness immediately after placement of a continuous cervical paravertebral block for postoperative pain control after shoulder stabilization surgery. The tip of the stimulating catheter was demonstrated in the C7 neuroforamen. Contrast injected through the catheter demonstrated epidural spread. The contralateral block resolved after 4 hours and the patient suffered no respiratory embarrassment or other untoward sequelae. CONCLUSION: Continuous cervical paravertebral block is a relatively new, but generally well-accepted, modality for postoperative pain control after major surgery to the upper limb. Epidural spread is recognized as a complication. In this particular case, medial placement of the catheter was possibly caused by unintentional medial direction of the bevel of the Tuohy needle. Meticulous attention to the direction of the needle bevel and early recognition and management of adverse events are mandatory. The same principles may apply for continuous thoracic, lumbar, and sacral paravertebral blocks. PMID- 16701199 TI - Relative performance of soft contact lenses having lathe-cut posterior surfaces with and without additional polishing. AB - After a preliminary investigation of the effects of tool feed rate and spindle speed on the surface roughness of unhydrated, lathe-cut polymacon surfaces, a laboratory and clinical comparison was made between lenses with identical parameters except that the lathe-cut posterior surface was left unpolished in the "test" lenses and was polished in the "control" lenses. The lenses had moulded anterior surfaces. Laboratory comparisons included surface roughness, lens power and its uniformity across the surface. Double-blind clinical trials over 4-hour (27 subjects) and 1-month (10 subjects) periods, involved one eye of each subject wearing a "test" lens and the other, a "control" lens. No clinically significant differences were found between the results for the test and control lenses. It is concluded that today's lathing technology makes a final polishing stage unnecessary. PMID- 16701200 TI - Under the eye of Nr-CAM. AB - Binocular vision relies upon the existence of contralateral and ispilateral projections from retinal ganglion cells. Contacts between visual axons and optic chiasm cells are critical for the sorting of crossed and uncrossed projections during development. In this issue of Neuron, a study by Williams et al. shows that the cell adhesion molecule Nr-CAM facilitates/promotes the decussation of contralateral axons across the chiasm. PMID- 16701201 TI - GABAB receptor isoforms caught in action at the scene. AB - The metabotropic GABAB receptors mediate slow synaptic inhibition and consist of heterodimers of GABAB1 and GABAB2 subunits. The only known molecular diversity of the GABAB receptors arises from the two GABAB1 isoforms, but its functional significance has been unclear. Two studies in this issue of Neuron now demonstrate that GABAB1a and GABAB1b show strategically distinct subcellular localization and physiological action. PMID- 16701202 TI - Seeing what the mouse sees with its vibrissae: a matter of behavioral state. AB - The behavioral state of an animal is accompanied by ongoing brain activity that primes neuronal circuitry to sensory inputs. While it should come as no surprise that the pattern of cortical activation is tied to behavioral states, only now has this dependence been imaged. In this issue of Neuron, Ferezou, Bolea, and Petersen show that the level and spatial extent of activation of vibrissa sensory cortex critically depend on behavioral context and mode of stimulation, i.e., passive versus active contact. PMID- 16701203 TI - Parkin blushed by PINK1. AB - Mutations in the PTEN-induced putative kinase 1 (PINK1) are a common cause of autosomal recessive Parkinson's disease. In a recent issue of Nature, two independent reports by and show that loss of Drosophila PINK1 leads to defects in mitochondrial function resulting in male sterility, apoptotic muscle degeneration, and minor loss of dopamine neurons that is rescued by overexpression of the ubiquitin E3 ligase, parkin. Thus, PINK1 and parkin appear to function in a common pathway suggesting a convergence of the two genes most commonly associated with autosomal recessive PD. PMID- 16701204 TI - The neural basis of mentalizing. AB - Mentalizing refers to our ability to read the mental states of other agents and engages many neural processes. The brain's mirror system allows us to share the emotions of others. Through perspective taking, we can infer what a person currently believes about the world given their point of view. Finally, the human brain has the unique ability to represent the mental states of the self and the other and the relationship between these mental states, making possible the communication of ideas. PMID- 16701205 TI - A role for Nr-CAM in the patterning of binocular visual pathways. AB - Retinal ganglion cell (RGC) axons diverge within the optic chiasm to project to opposite sides of the brain. In mouse, contralateral RGCs are distributed throughout the retina, whereas ipsilateral RGCs are restricted to the ventrotemporal crescent (VTC). While repulsive guidance mechanisms play a major role in the formation of the ipsilateral projection, little is known about the contribution of growth-promoting interactions to the formation of binocular visual projections. Here, we show that the cell adhesion molecule Nr-CAM is expressed by RGCs that project contralaterally and is critical for the guidance of late-born RGCs within the VTC. Blocking Nr-CAM function causes an increase in the size of the ipsilateral projection and reduces neurite outgrowth on chiasm cells in an age- and region-specific manner. Finally, we demonstrate that EphB1/ephrin-B2-mediated repulsion and Nr-CAM-mediated attraction comprise distinct molecular programs that each contributes to the proper formation of binocular visual pathways. PMID- 16701206 TI - Cell survival through Trk neurotrophin receptors is differentially regulated by ubiquitination. AB - Specificity of neurotrophin factor signaling is dictated through the action of Trk receptor tyrosine kinases. Once activated, Trk receptors are internalized and targeted for degradation. However, the mechanisms implicated in this process are incompletely understood. Here we report that the Trk receptors are multimonoubiquitinated in response to neurotrophins. We have identified an E3 ubiquitin ligase, Nedd4-2, that associates with the TrkA receptor and is phosphorylated upon NGF binding. The binding of Nedd4-2 to TrkA through a PPXY motif leads to the ubiquitination and downregulation of TrkA. Activated TrkA receptor levels and the survival of NGF-dependent sensory neurons, but not BDNF dependent sensory neurons, are directly influenced by Nedd4-2 expression. Unexpectedly, Nedd4-2 does not bind or ubiquitinate related TrkB receptors, due to the lack of a consensus PPXY motif. Our results indicate that Trk neurotrophin receptors are differentially regulated by ubiquitination to modulate the survival of neurons. PMID- 16701207 TI - Modulation of basal and receptor-induced GIRK potassium channel activity and neuronal excitability by the mammalian PINS homolog LGN. AB - G protein-activated inwardly rectifying potassium (GIRK) channels mediate slow synaptic inhibition and control neuronal excitability. It is unknown whether GIRK channels are subject to regulation by guanine dissociation inhibitor (GDI) proteins like LGN, a mammalian homolog of Drosophila Partner of Inscuteable (mPINS). Here we report that LGN increases basal GIRK current but reduces GIRK activation by metabotropic transmitter receptors coupled to Gi or Go, but not Gs. Moreover, expression of its N-terminal, TPR-containing protein interaction domains mimics the effects of LGN in mammalian cells, probably by releasing sequestered endogenous LGN. In hippocampal neurons, expression of LGN, or LGN fragments that mimic or enhance LGN activity, hyperpolarizes the resting potential due to increased basal GIRK activity and reduces excitability. Using Lenti virus for LGN RNAi to reduce endogenous LGN levels in hippocampal neurons, we further show an essential role of LGN for maintaining basal GIRK channel activity and for harnessing neuronal excitability. PMID- 16701208 TI - A shared vesicular carrier allows synaptic corelease of GABA and glycine. AB - The type of vesicular transporter expressed by a neuron is thought to determine its neurotransmitter phenotype. We show that inactivation of the vesicular inhibitory amino acid transporter (Viaat, VGAT) leads to embryonic lethality, an abdominal defect known as omphalocele, and a cleft palate. Loss of Viaat causes a drastic reduction of neurotransmitter release in both GABAergic and glycinergic neurons, indicating that glycinergic neurons do not express a separate vesicular glycine transporter. This loss of GABAergic and glycinergic synaptic transmission does not impair the development of inhibitory synapses or the expression of KCC2, the K+ -Cl- cotransporter known to be essential for the establishment of inhibitory neurotransmission. In the absence of Viaat, GABA-synthesizing enzymes are partially lost from presynaptic terminals. Since GABA and glycine compete for vesicular uptake, these data point to a close association of Viaat with GABA synthesizing enzymes as a key factor in specifying GABAergic neuronal phenotypes. PMID- 16701210 TI - The GABAB1b isoform mediates long-lasting inhibition of dendritic Ca2+ spikes in layer 5 somatosensory pyramidal neurons. AB - The apical tuft of layer 5 pyramidal neurons is innervated by a large number of inhibitory inputs with unknown functions. Here, we studied the functional consequences and underlying molecular mechanisms of apical inhibition on dendritic spike activity. Extracellular stimulation of layer 1, during blockade of glutamatergic transmission, inhibited the dendritic Ca2+ spike for up to 400 ms. Activation of metabotropic GABAB receptors was responsible for a gradual and long-lasting inhibitory effect, whereas GABAA receptors mediated a short-lasting (approximately 150 ms) inhibition. Our results suggest that the mechanism underlying the GABAB inhibition of Ca2+ spikes involves direct blockade of dendritic Ca2+ channels. By using knockout mice for the two predominant GABAB1 isoforms, GABAB1a and GABAB1b, we showed that postsynaptic inhibition of Ca2+ spikes is mediated by GABAB1b, whereas presynaptic inhibition of GABA release is mediated by GABAB1a. We conclude that the molecular subtypes of GABAB receptors play strategically different physiological roles in neocortical neurons. PMID- 16701209 TI - Differential compartmentalization and distinct functions of GABAB receptor variants. AB - GABAB receptors are the G protein-coupled receptors for the main inhibitory neurotransmitter in the brain, gamma-aminobutyric acid (GABA). Molecular diversity in the GABAB system arises from the GABAB1a and GABAB1b subunit isoforms that solely differ in their ectodomains by a pair of sushi repeats that is unique to GABAB1a. Using a combined genetic, physiological, and morphological approach, we now demonstrate that GABAB1 isoforms localize to distinct synaptic sites and convey separate functions in vivo. At hippocampal CA3-to-CA1 synapses, GABAB1a assembles heteroreceptors inhibiting glutamate release, while predominantly GABAB1b mediates postsynaptic inhibition. Electron microscopy reveals a synaptic distribution of GABAB1 isoforms that agrees with the observed functional differences. Transfected CA3 neurons selectively express GABAB1a in distal axons, suggesting that the sushi repeats, a conserved protein interaction motif, specify heteroreceptor localization. The constitutive absence of GABAB1a but not GABAB1b results in impaired synaptic plasticity and hippocampus-dependent memory, emphasizing molecular differences in synaptic GABAB functions. PMID- 16701211 TI - Visualizing the cortical representation of whisker touch: voltage-sensitive dye imaging in freely moving mice. AB - Voltage-sensitive dye imaging resolves the spatiotemporal dynamics of supragranular subthreshold cortical activity with millisecond temporal resolution and subcolumnar spatial resolution. We used a flexible fiber optic image bundle to visualize voltage-sensitive dye dynamics in the barrel cortex of freely moving mice while simultaneously filming whisker-related behavior to generate two movies matched frame-by-frame with a temporal resolution of up to 2 ms. Sensory responses evoked by passive whisker stimulation lasted longer and spread further across the barrel cortex in awake mice compared to anesthetized mice. Passively evoked sensory responses were large during behaviorally quiet periods and small during active whisking. However, as an exploring mouse approached an object while whisking, large-amplitude, propagating cortical sensory activity was evoked by active whisker-touch. These experiments demonstrate that fiber optics can be used to image cortical sensory activity with high resolution in freely moving animals. The results demonstrate differential processing of sensory input depending upon behavior. PMID- 16701212 TI - Activity in the lateral prefrontal cortex reflects multiple steps of future events in action plans. AB - To achieve a behavioral goal in a complex environment, we must plan multiple steps of motor behavior. On planning a series of actions, we anticipate future events that will occur as a result of each action and mentally organize the temporal sequence of events. To investigate the involvement of the lateral prefrontal cortex (PFC) in such multistep planning, we examined neuronal activity in the PFC of monkeys performing a maze task that required the planning of stepwise cursor movements to reach a goal. During the preparatory period, PFC neurons reflected each of all forthcoming cursor movements, rather than arm movements. In contrast, in the primary motor cortex, most neuronal activity reflected arm movements but little of cursor movements during the preparatory period, as well as during movement execution. Our data suggest that the PFC is involved primarily in planning multiple future events that occur as a consequence of behavioral actions. PMID- 16701213 TI - Anterior cingulate and posterior parietal cortices are sensitive to dissociable forms of conflict in a task-switching paradigm. AB - The conflict-monitoring hypothesis posits that anterior cingulate cortex (ACC) monitors conflict in information processing and recruits dorsolateral prefrontal cortex (DLPFC) to resolve competition as needed. We used fMRI to test this prediction directly in the context of a task-switching paradigm, in which subjects responded to the color or the motion of a visual stimulus. Conflict was indexed in terms of the product of activities in areas specialized for color or motion processing on a trial-by-trial basis. Here, we report that ACC and posterior parietal cortex (PPC) were sensitive to distinct forms of conflict, at the level of the response and the stimulus representation, respectively. Activity in PPC preceded increased activity in DLPFC and predicted enhanced behavioral performance on subsequent trials. These findings suggest that ACC and PPC may act in concert to detect dissociable forms of conflict and signal to DLPFC the need for increased control. PMID- 16701214 TI - Dissociable medial prefrontal contributions to judgments of similar and dissimilar others. AB - Human social interaction requires the recognition that other people are governed by the same types of mental states-beliefs, desires, intentions-that guide one's own behavior. We used functional neuroimaging to examine how perceivers make mental state inferences when such self-other overlap can be assumed (when the other is similar to oneself) and when it cannot (when the other is dissimilar from oneself). We observed a double dissociation such that mentalizing about a similar other engaged a region of ventral mPFC linked to self-referential thought, whereas mentalizing about a dissimilar other engaged a more dorsal subregion of mPFC. The overlap between judgments of self and similar others suggests the plausibility of "simulation" accounts of social cognition, which posit that perceivers can use knowledge about themselves to infer the mental states of others. PMID- 16701215 TI - Molecular basis of sex determination in haplodiploids. AB - Sex in many species of Hymenoptera (ants, bees and wasps) is determined by a single locus that is heterozygous in females and hemizygous in (haploid) males. Beye and colleagues have now cloned the csd locus in the honeybee Apis mellifera and provide functional evidence that this gene is the primary switch in the sex determination cascade of honeybees and possibly all Hymenoptera. PMID- 16701216 TI - Are men and women really so different? AB - Distinct differences in the behaviour and preferences of men and women have conventionally been attributed to Trivers' powerful insights regarding the impact of parental investment on sexual selection and mating systems. This has spawned a huge literature about the evolutionary significance of human sex differences. But are men and women really so different? An elegant new study shows that men and women are strikingly similar in their mate preferences. Have conventional models blinded us to the obvious, and precluded the posing of far more interesting questions? PMID- 16701217 TI - Nature is changing in more ways than one. PMID- 16701219 TI - Long-lived fellows. PMID- 16701220 TI - Conserving tropical nature: current challenges for ecologists. AB - Tropical biodiversity continues to erode unabated, which calls for ecologists to address the problem directly, placing less reliance on indirect interventions, such as community-based development schemes. Ecologists must become more assertive in providing scientifically formulated and adaptively managed interventions, involving biodiversity payments, to serve local, regional and global interests in tropical nature. Priorities for tropical ecologists thus include the identification of key thresholds to ecological resilience, and the formulation of clear monitoring protocols and management strategies for implementation by local resource managers. A particular challenge is to demonstrate how nature reserves contribute to the adaptive capacity of regional land-use matrices and, hence, to the provision of sustainable benefits at multiple spatial and temporal scales. PMID- 16701221 TI - Ecological and evolutionary consequences of biotic homogenization. AB - Biotic homogenization, the gradual replacement of native biotas by locally expanding non-natives, is a global process that diminishes floral and faunal distinctions among regions. Although patterns of homogenization have been well studied, their specific ecological and evolutionary consequences remain unexplored. We argue that our current perspective on biotic homogenization should be expanded beyond a simple recognition of species diversity loss, towards a synthesis of higher order effects. Here, we explore three distinct forms of homogenization (genetic, taxonomic and functional), and discuss their immediate and future impacts on ecological and evolutionary processes. Our goal is to initiate future research that investigates the broader conservation implications of homogenization and to promote a proactive style of adaptive management that engages the human component of the anthropogenic blender that is currently mixing the biota on Earth. PMID- 16701222 TI - Visual perception and social foraging in birds. AB - Birds gather information about their environment mainly through vision by scanning their surroundings. Many prevalent models of social foraging assume that foraging and scanning are mutually exclusive. Although this assumption is valid for birds with narrow visual fields, these models have also been applied to species with wide fields. In fact, available models do not make precise predictions for birds with large visual fields, in which the head-up, head-down dichotomy is not accurate and, moreover, do not consider the effects of detection distance and limited attention. Studies of how different types of visual information are acquired as a function of body posture and of how information flows within flocks offer new insights into the costs and benefits of living in groups. PMID- 16701223 TI - Genome diversity in microbial eukaryotes. AB - The genomic peculiarities among microbial eukaryotes challenge the conventional wisdom of genome evolution. Currently, many studies and textbooks explore principles of genome evolution from a limited number of eukaryotic lineages, focusing often on only a few representative species of plants, animals and fungi. Increasing emphasis on studies of genomes in microbial eukaryotes has and will continue to uncover features that are either not present in the representative species (e.g. hypervariable karyotypes or highly fragmented mitochondrial genomes) or are exaggerated in microbial groups (e.g. chromosomal processing between germline and somatic nuclei). Data for microbial eukaryotes have emerged from recent genome sequencing projects, enabling comparisons of the genomes from diverse lineages across the eukaryotic phylogenetic tree. Some of these features, including amplified rDNAs, subtelomeric rDNAs and reduced genomes, appear to have evolved multiple times within eukaryotes, whereas other features, such as absolute strand polarity, are found only within single lineages. PMID- 16701224 TI - Transients: the key to long-term ecological understanding? AB - Ecological theory has been dominated by a focus on long-term or asymptotic behavior as a way to understand natural systems. Yet experiments are done on much shorter timescales, and the relevant timescales for ecological systems can also be relatively short. Thus, there is a mismatch between the timescales of most experiments and the timescales of many theoretical investigations. However, recent work has emphasized the importance of transient dynamics rather than long term behavior in ecological systems, enabling the examination of forces that allow coexistence on ecological timescales. Through an examination of what leads to transients in ecological systems, a deeper appreciation of the forces leading to persistence or coexistence in ecological systems emerges, as well as a general understanding of how population levels can change through time. PMID- 16701225 TI - Alternative states and positive feedbacks in restoration ecology. AB - There is increasing interest in developing better predictive tools and a broader conceptual framework to guide the restoration of degraded land. Traditionally, restoration efforts have focused on re-establishing historical disturbance regimes or abiotic conditions, relying on successional processes to guide the recovery of biotic communities. However, strong feedbacks between biotic factors and the physical environment can alter the efficacy of these successional-based management efforts. Recent experimental work indicates that some degraded systems are resilient to traditional restoration efforts owing to constraints such as changes in landscape connectivity and organization, loss of native species pools, shifts in species dominance, trophic interactions and/or invasion by exotics, and concomitant effects on biogeochemical processes. Models of alternative ecosystem states that incorporate system thresholds and feedbacks are now being applied to the dynamics of recovery in degraded systems and are suggesting ways in which restoration can identify, prioritize and address these constraints. PMID- 16701226 TI - Cooperate or speciate: new theory for the distribution of passerine birds. AB - In cooperatively breeding birds, adults often forego reproduction and help care for the offspring of others. A universal explanation for this mode of breeding has eluded evolutionary biologists, who have considered it to be a rare, and largely Australian, phenomenon. In a recent paper, Andrew Cockburn reports that the number of known cooperative breeders among oscine passerine birds has more than doubled since the last substantial review, published 16 years ago. Cooperative breeding is often the ancestral trait, and predominantly cooperative genera are species poor compared with their pair-breeding counterparts. Cockburn argues that speciation is less likely in cooperative clades, because the philopatric tendencies of individuals make them poor dispersers, colonizers and migrants. This new hypothesis helps explain the distribution and composition of migrant and island avifauna. However, a major challenge remains to reconcile the roles of phylogenetic history and current ecology in promoting cooperative behaviour. PMID- 16701227 TI - Ecological and evolutionary implications of immunological priming in invertebrates. AB - Invertebrates have an immune response that differs considerably from the acquired immune response found in vertebrates. However, new studies indicate that past experience with a pathogen can provide individual invertebrates, or their descendants, with enhanced immunity. This prophylactic effect, termed immunological priming, is functionally similar to the acquired immune response in vertebrates. This newfound complexity of invertebrate immunity begs investigation into the conditions under which immunological priming should evolve, and its consequences for population dynamics. PMID- 16701230 TI - Human expansion: the roles of castaways and cultural practices. PMID- 16701228 TI - Biological control and indirect effects. PMID- 16701231 TI - Evolving new genetic codes. AB - Although the genetic code is almost universal, natural variations exist that have caused evolutionary biologists to speculate about codon evolution. There are two predominant hypotheses that specify either a gradual (ambiguous intermediate) or stochastic (codon capture) change in the code. These hypotheses are similar to two biotechnology techniques that have been used to engineer the genetic code: a 'top down' approach, in which the whole organism is evolved for the ability to incorporate unnatural amino acids, and a 'bottom up' approach, in which aminoacyl tRNA synthetases and their cognate tRNAs are engineered. The biotechnology experiments provide insights into natural codon evolution, and a combination of these approaches should enable the evolution of organisms that can incorporate unnatural amino acids throughout their proteomes. PMID- 16701232 TI - Genetic details, optimization and phage life histories. AB - Optimality models assume that phenotypes evolve by natural selection largely independently of underlying genetic mechanisms. This neglect of genetic mechanisms is considered an advantage by some evolutionary biologists but a fatal flaw by others. The controversy has gone unresolved, in part, from a lack of complex phenotypes that meet optimality criteria and for which the underlying genetic mechanisms are known. Here, we look at both perspectives for lysis time in bacteriophages. We find that the basic assumptions of the optimality model are compatible with the genetic details, but the optimality model is limited in its ability to accommodate lysis time plasticity because the mechanistic underpinnings of plasticity are poorly known. PMID- 16701233 TI - A major evolutionary transition to more than two sexes? AB - Two recently discovered cases of genetic caste determination in social insects might provide the first example of a major evolutionary transition from two to more than two sexes. I argue here that the system can be interpreted as comprising primarily individuals requiring gametes from three parental types and having four sexes from the perspective of demographic extinction. Additionally, I show how this mating system can be seen as a major evolutionary transition. For these populations, it is apparent that the mechanism for a three- or four-sex system does not lie within the myriad of possible arrangements of chromosomes within individuals, but at the next level of evolutionary complexity, with the arrangement of chromosomes among individuals within a social system. PMID- 16701234 TI - Sexual selection and genital evolution. AB - Genitalia are conspicuously variable, even in closely related taxa that are otherwise morphologically very similar. Explaining genital diversity is a longstanding problem that is attracting renewed interest from evolutionary biologists. New studies provide ever more compelling evidence that sexual selection is important in driving genital divergence. Importantly, several studies now link variation in genital morphology directly to male fertilization success, and modern comparative techniques have confirmed predicted associations between genital complexity and mating patterns across species. There is also evidence that male and female genitalia can coevolve antagonistically. Determining mechanisms of genital evolution is an important challenge if we are to resolve current debate concerning the relative significance of mate choice benefits and sexual conflict in sexual selection. PMID- 16701235 TI - Adaptation to natural flow regimes. AB - Floods and droughts are important features of most running water ecosystems, but the alteration of natural flow regimes by recent human activities, such as dam building, raises questions related to both evolution and conservation. Among organisms inhabiting running waters, what adaptations exist for surviving floods and droughts? How will the alteration of the frequency, timing and duration of flow extremes affect flood- and drought-adapted organisms? How rapidly can populations evolve in response to altered flow regimes? Here, we identify three modes of adaptation (life history, behavioral and morphological) that plants and animals use to survive floods and/or droughts. The mode of adaptation that an organism has determines its vulnerability to different kinds of flow regime alteration. The rate of evolution in response to flow regime alteration remains an open question. Because humans have now altered the flow regimes of most rivers and many streams, understanding the link between fitness and flow regime is crucial for the effective management and restoration of running water ecosystems. PMID- 16701236 TI - Model selection in ecology and evolution. AB - Recently, researchers in several areas of ecology and evolution have begun to change the way in which they analyze data and make biological inferences. Rather than the traditional null hypothesis testing approach, they have adopted an approach called model selection, in which several competing hypotheses are simultaneously confronted with data. Model selection can be used to identify a single best model, thus lending support to one particular hypothesis, or it can be used to make inferences based on weighted support from a complete set of competing models. Model selection is widely accepted and well developed in certain fields, most notably in molecular systematics and mark-recapture analysis. However, it is now gaining support in several other areas, from molecular evolution to landscape ecology. Here, we outline the steps of model selection and highlight several ways that it is now being implemented. By adopting this approach, researchers in ecology and evolution will find a valuable alternative to traditional null hypothesis testing, especially when more than one hypothesis is plausible. PMID- 16701237 TI - Tits, noise and urban bioacoustics. AB - Humans, particularly in cities, are noisy. Researchers are only just beginning to identify the implications of an increase in noise for species that communicate acoustically. In a recent paper, Slabbekoorn and Peet show, for the first time, that some birds can respond to anthropogenically elevated noise levels by altering the frequency structure of their songs. Cities are fruitful grounds for research on the evolution of animal communication systems, with broader implications for conservation in human-altered environments. PMID- 16701238 TI - Speciation in the apple maggot fly: a blend of vintages? AB - The importance of speciation without the complete geographical separation of diverging populations (sympatric speciation) has become increasingly accepted. One of the textbook examples of recent speciation in sympatry is the apple maggot fly Rhagoletis pomonella, in which genetically differentiated host races feed on either hawthorn or apple. Three recent articles by Feder and collaborators show that the history of these host races is more complicated than was previously realized. Genes that differentiate races of flies that feed on either apple or hawthorn are located in chromosomal rearrangements. This variation forms a latitudinal cline that must have been established long before apples were available as hosts. Furthermore, there is also new evidence for the very recent evolution of a derived preference for volatile chemicals that are typical of apple fruits among apple-feeding flies. These results show that adaptation to apple populations has involved both the sorting of ancestral adaptive variation and the selection of novel mutations. PMID- 16701239 TI - What is right with 'teaching the controversy'? PMID- 16701240 TI - Teaching about scientific dissent from neo-darwinism. PMID- 16701242 TI - Rules of thumb for judging ecological theories. AB - An impressive fit to historical data suggests to biologists that a given ecological model is highly valid. Models often achieve this fit at the expense of exaggerated complexity that is not justified by empirical evidence. Because overfitted theories complement the traditional assumption that ecology is 'messy', they generally remain unquestioned. Using predation theory as an example, we suggest that a fit-driven appraisal of model value is commonly misdirected; although fit to historical data can be important, the simplicity and generality of a theory--and thus its ecological value--are of comparable importance. In particular, we argue that theories whose complexity greatly exceeds the complexity of the problem that they address should be rejected. We suggest heuristics for distinguishing between valuable ecological theories and their overfitted brethren. PMID- 16701243 TI - Promiscuous females protect their offspring. AB - Multi-male mating (MMM) by females is relatively common among mammals, occurring in at least 133 species and several evolutionary benefits of MMM have been proposed. The most convincing explanation is that MMM confuses paternity, thereby deterring infanticide by males. A second explanation for females that are unlikely to experience infanticide is that MMM is a consequence of sexual harassment. Mate guarding and, perhaps even in some cases, behavioral monogamy, might have evolved in response to the threat of infanticide and the subsequent tendency for females to mate multiply. Benefits relating to improved genetic fitness of offspring do occur in some species, but do not provide a widespread explanation for the evolutionary origin of MMM; if cryptic female choice through sperm competition is adaptive to females it probably evolved as a consequence of, rather than a precursor to, female promiscuity. Here, we provide support for the original hypothesis of paternity confusion for MMM, rather than for the more popular good genes or sperm competition hypotheses. PMID- 16701244 TI - What is altruism? AB - Altruism is generally understood to be behavior that benefits others at a personal cost to the behaving individual. However, within evolutionary biology, different authors have interpreted the concept of altruism differently, leading to dissimilar predictions about the evolution of altruistic behavior. Generally, different interpretations diverge on which party receives the benefit from altruism and on how the cost of altruism is assessed. Using a simple trait-group framework, we delineate the assumptions underlying different interpretations and show how they relate to one another. We feel that a thorough examination of the connections between interpretations not only reveals why different authors have arrived at disparate conclusions about altruism, but also illuminates the conditions that are likely to favor the evolution of altruism. PMID- 16701245 TI - Isolation of nucleic acids and cultures from fossil ice and permafrost. AB - Owing to their constant low temperatures, glacial ice and permafrost might contain the oldest nucleic acids and microbial cells on Earth, which could prove key to reconstructing past ecosystems and for the planning of missions to other planets. However, recent claims concerning viable cells and microbial nucleic acids obtained from ice- and permafrost cores from hundreds of thousands to millions of years old are not properly authenticated and the findings could be the result of contamination. Here, we discuss the processes that restrict the long-term survival of DNA and/or RNA molecules in ice and permafrost, and highlight sources of contamination that could result in false claims. Additionally, we present a set of precautions, controls and criteria to help ensure that future cultures and sequences are authentic. PMID- 16701246 TI - Carbon isotopes in functional soil ecology. AB - Soil is an integral part of terrestrial ecosystems. Many soil ecologists interested in soil ecosystem functioning rely, to some degree, on stable isotope methodologies. The study of the natural abundance of carbon isotopes, especially (13)C but also (14)C, in the environment and the use of stable carbon isotope tracers have proved very useful in investigating the soil carbon cycle and soil trophic relationships. Recent methodological and technical advances have greatly extended the possibilities for the application of stable carbon isotopes to terrestrial ecology and have vastly improved our knowledge of belowground ecosystem functioning and will continue to do so. A better understanding of soil processes is invaluable in predicting the future impacts of global environmental change on terrestrial ecosystems. PMID- 16701247 TI - Diplochory: are two seed dispersers better than one? AB - Diplochory is seed dispersal by a sequence of two or more steps or phases, each involving a different dispersal agent. Here, we describe five forms of diplochory and derive general characteristics of each phase of seed dispersal. The first and second phases of diplochory offer different benefits to plants. Phase one dispersal often results in escape from density-dependent seed and seedling mortality near the parent plant and can result in the colonization of habitat patches far from the parent. Phase two dispersal often moves seeds to discrete and predictable microsites, where the probability of seedling establishment is disproportionately high (i.e. directed dispersal). Diplochory appears to have evolved because combining two means of seed dispersal can increase the benefits of seed dispersal whilst reducing the likelihood of seed mortality. PMID- 16701248 TI - Are filarial nematode Wolbachia obligate mutualist symbionts? AB - The intracellular symbiotic bacteria of filarial nematodes have inspired new ideas for the control of disease using antibacterial drugs. For effective, long term control, this requires that the bacteria are essential to their nematode hosts. Two recent studies offer conflicting evidence: long, close coevolution between most filarial nematodes and their symbionts contrasts with many species having naturally lost them. An attempt to transfer symbionts to an uninfected host found that the bacteria did not thrive, suggesting they are adapted to one host. PMID- 16701249 TI - Allelic histories: positive selection on a HIV-resistance allele. AB - The CCR5-Delta32 allele crucially determines the course of HIV infection and appears to be highly protective against the disease. Population genetic studies suggest that the allele has been under positive selection in Europe in the past. In a recent paper, Alison Galvani and Montgomery Slatkin collate the available evidence and use a mathematical model to strongly suggest that smallpox could have exerted sufficient selection pressure to explain the distribution of the allele across Europe. This is a beautiful example of the power of mathematical models in evolutionary genetics. PMID- 16701251 TI - Reflections on animal selves. AB - Is self-cognizance a uniquely human attribute, or do other animals also have a sense of self? Although there is considerable interest in this question, answers remain elusive. Progress has been stymied by misunderstandings in terminology, a focus on a narrow range of species, and controversies over key concepts, experimental paradigms and interpretations of data. Here, we propose a new conceptual and terminological framework, emphasizing that degrees of self cognizance differ among animals because of the cognitive demands that their species-specific social structures and life-history characteristics have placed upon them over evolutionary time. We suggest that the self-cognizance of an organism falls at a point on a continuum of social complexity and conscious involvement. PMID- 16701250 TI - Democracy in animal groups: a political science perspective. PMID- 16701252 TI - Ruminating on complexity: macroparasites of wildlife and livestock. AB - Recent advances in ecology have improved our understanding of the role of parasites in the dynamics of wildlife populations. However, conditions that prevail in many wildlife systems, such as host movement, contact with livestock, and heterogeneity in the environment of the parasite outside of the host, have largely been ignored in existing models of macroparasite transmission. We need to refine these models if we are to stand a chance of developing effective parasite control strategies. New quantitative approaches enable us to address key complexities and make better use of scarce data, and these should enhance our efforts to understand and control emerging problems of interspecific parasite transmission. PMID- 16701254 TI - Hybridization and adaptive radiation. AB - Whether interspecific hybridization is important as a mechanism that generates biological diversity is a matter of controversy. Whereas some authors focus on the potential of hybridization as a source of genetic variation, functional novelty and new species, others argue against any important role, because reduced fitness would typically render hybrids an evolutionary dead end. By drawing on recent developments in the genetics and ecology of hybridization and on principles of ecological speciation theory, I develop a concept that reconciles these views and adds a new twist to this debate. Because hybridization is common when populations invade new environments and potentially elevates rates of response to selection, it predisposes colonizing populations to rapid adaptive diversification under disruptive or divergent selection. I discuss predictions and suggest tests of this hybrid swarm theory of adaptive radiation and review published molecular phylogenies of adaptive radiations in light of the theory. PMID- 16701253 TI - Big questions, small worlds: microbial model systems in ecology. AB - Although many biologists have embraced microbial model systems as tools to address genetic and physiological questions, the explicit use of microbial communities as model systems in ecology has traditionally been more restricted. Here, we highlight recent studies that use laboratory-based microbial model systems to address ecological questions. Such studies have significantly advanced our understanding of processes that have proven difficult to study in field systems, including the genetic and biochemical underpinnings of traits involved in ecological interactions, and the ecological differences driving evolutionary change. It is the simplicity of microbial model systems that makes them such powerful tools for the study of ecology. Such simplicity enables the high degrees of experimental control and replication that are necessary to address many questions that are inaccessible through field observation or experimentation. PMID- 16701255 TI - A general law for animal locomotion? AB - The propulsion system of animals that fly or swim are quite different from each other in their morphology and function, yet the propulsive efficiency could be maximized by a surprising similarity in the fine tuning of flapping frequency, amplitude and forward speed, according to a new study by Taylor et al. This conclusion was based on an analysis of the Strouhal number, which is a dynamic similarity index relevant to propulsion that relies on vortex shedding for thrust generation. Such fine-tuning of the propulsive system suggests possible consequences for physiological and ecological adaptations related to, for example muscle operating frequency and optimal speed of muscle contraction. PMID- 16701256 TI - Disease dynamics: all caused by males? AB - Some host individuals tend to acquire parasites at a much faster rate than do others--a consequence of heterogeneities in susceptibility and/or exposure. This is termed 'overdispersion' and, as for many other statistical phenomena, the degree of overdispersion often conforms to a 20/80 rule, where 20% of the host population is responsible for approximately 80% of the parasite transmission. But which are the hosts driving so much of the dynamics of an infectious disease? If host individuals at the tail of the frequency distribution can be identified by some common label, controlling parasitic diseases would be much easier. In two recent papers, Perkins et al. and Ferrari et al. have shown that male hosts are much more important than female hosts in the transmission of parasites. PMID- 16701257 TI - The millennium ecosystem assessment: what is it all about? AB - Hundreds of scientists from over 70 nations are now engaged in an intensive effort to assess what we know about the status of the world's ecosystems. Here, we describe the fundamental nature of this assessment, what it hopes to accomplish and how it will go about its work. The results of the Millennium Ecosystem Assessment will serve as a baseline for future assessments, as well as a blueprint for action for sustaining the ecosystem services upon which we all depend for our well being. PMID- 16701258 TI - Sexual selection and animal genitalia. PMID- 16701259 TI - Conserving tropical biodiversity: the arid end of the scale. PMID- 16701261 TI - Is community-based ecotourism a good use of biodiversity conservation funds? AB - Community-based ecotourism (CBET) has become a popular tool for biodiversity conservation, based on the principle that biodiversity must pay for itself by generating economic benefits, particularly for local people. There are many examples of projects that produce revenues for local communities and improve local attitudes towards conservation, but the contribution of CBET to conservation and local economic development is limited by factors such as the small areas and few people involved, limited earnings, weak linkages between biodiversity gains and commercial success, and the competitive and specialized nature of the tourism industry. Many CBET projects cited as success stories actually involve little change in existing local land and resource-use practices, provide only a modest supplement to local livelihoods, and remain dependent on external support for long periods, if not indefinitely. Investment in CBET might be justified in cases where such small changes and benefits can yield significant conservation and social benefits, although it must still be recognized as requiring a long term funding commitment. Here, I aim to identify conditions under which CBET is, and is not, likely to be effective, efficient and sustainable compared with alternative approaches for conserving biodiversity. I also highlight the need for better data and more rigorous analysis of both conservation and economic impacts. PMID- 16701262 TI - Mother's curse: the effect of mtDNA on individual fitness and population viability. AB - The mitochondrial genome is considered generally to be an innocent bystander in adaptive evolution; however, there is increasing evidence that mitochondrial DNA (mtDNA) is an important contributor to viability and fecundity. Some of this evidence is now well documented, with mtDNA mutations having been shown to play a causal role in degenerative diseases, ageing, and cancer. However, most research on mtDNA has ignored the possibility that other instances exist where mtDNA mutations could have profound fitness consequences. Recent work in humans and other species now indicates that mtDNA mutations play an important role in sperm function, male fertility, and male fitness. Ironically, deleterious mtDNA mutations that affect only males, such as those that impair sperm function, will not be subject to natural selection because mitochondria are generally maternally inherited and could reach high frequencies in populations if the mutations are not disadvantageous in females. Here, we review how such mtDNA mutations might affect the viability of natural populations. We consider factors that increase or decrease the strength of the effect of mtDNA mutations on population viability and discuss what mechanisms exist to mitigate deleterious mtDNA effects. PMID- 16701263 TI - What is wrong with absolute individual fitness? AB - One of the most basic facts about evolution is that fitness is a relative concept. It does not matter how well an organism survives and reproduces, only that it does so better than other organisms bearing alternative traits. Nevertheless, many evolutionary arguments are framed in terms of absolute individual fitness. The absolute fitness criterion (AFC) can be justified in terms of relative fitness only given certain assumptions that are frequently violated in nature. In particular, interactions must occur in groups that are randomly formed and phenotypic variation among groups must be tightly coupled to genetic variation. Complicating the genotype-phenotype relationship can cause phenotypic variation among groups to become nonrandom, even when the groups are randomly formed, favoring traits that do not maximize absolute individual fitness. Complex genotype-phenotype relationships and complex population structures require explicit models of evolutionary change based on relative fitness differences within and among groups. PMID- 16701264 TI - Physiological stress in ecology: lessons from biomedical research. AB - Increasingly, levels of the 'stress hormones' cortisol and corticosterone are being used by ecologists as indicators of physiological stress in wild vertebrates. The amplitude of hormonal response is assumed to correlate with the overall health of an animal and, by extension, the health of the population. However, much of what is known about the physiology of stress has been elucidated by the biomedical research community. I summarize five physiological mechanisms that regulate hormone release during stress that should be useful to ecologists and conservationists. Incorporating these physiological mechanisms into the design and interpretation of ecological studies will make these increasingly popular studies of stress in ecological settings more rigorous. PMID- 16701265 TI - From birds to butterflies: animal movement patterns and stable isotopes. AB - Establishing patterns of movement of wild animals is crucial for our understanding of their ecology, life history and behavior, and is a prerequisite for their effective conservation. Advances in the use of stable isotope markers make it possible to track a diversity of animal species in a variety of habitats. This approach is revolutionizing the way in which we make connections between phases of the annual cycle of migratory animals. However, researchers must exercise care in their application of isotopic methods. Here, we review stable isotope patterns in nature and discuss recent tracking applications in a range of taxa. To aid in the interpretation and design of effective and insightful isotope movement studies, we discuss a series of key issues and assumptions. This exciting field will advance rapidly if researchers consider these aspects of study design and interpretation carefully. PMID- 16701266 TI - Quantitative genetic models of sexual selection. AB - Modeling of R.A. Fisher's ideas about the evolution of male ornamentation using quantitative genetics began in the 1980s. Following an initial period of enthusiasm, interest in these models began to wane when theoretical studies seemed to show that the rapid evolution of ornaments would not occur if there were costs associated with female mate choice. Recent theoretical work has shown, however, that runaway evolution and other kinds of extensive diversification of ornaments and preferences can occur, even when female choice is costly. These new models highlight crucial parameters that profoundly influence evolutionary trajectories, but these parameters have been neglected in empirical studies. Here, we review quantitative genetic models of sexual selection with the aim of fostering communication and synergism between theoretical and empirical enterprises. We also point out several areas in which additional empirical work could distinguish between alternative models of evolution. PMID- 16701268 TI - Two stressors are far deadlier than one. AB - Natural organisms often face a barrage of stressors, both natural and human induced. Two known stressors that impact amphibian populations are pesticides and predators. Recent work by Relyea and by Mills and Relyea reveals strikingly strong, synergistic negative effects of these two factors on amphibian larvae. Adding predation risk on top of supposedly sublethal concentrations of a common pesticide caused a massive increase in larval mortality. Interestingly, the increased mortality did not require exposure to actual predation. That is, simply the 'smell of danger' (predator chemical cues) caused 80-90% of larvae that were held in otherwise 'safe' levels of the pesticide to die. Notably, this effect occurred in some species, but not in others. These new studies highlight the need for further interdisciplinary work on the conditions under which combinations of stressors have particularly strong negative effects on natural organisms. PMID- 16701269 TI - Sustainability in a nutshell. AB - Sustainable exploitation is widely advocated as a strategy for reconciling economic pressures upon natural habitats with nature conservation. Two recent papers examine different aspects of the sustainability of the nut harvest on wild populations of Brazil nut trees Bertholletia excelsa in Amazonia. Peres et al. find that many populations of the Brazil nut tree lack juvenile trees and are not regenerating. In a socioeconomic study, Escobal and Aldana find that nut gathering provides insufficient income on its own to support nut-gatherers and that their other income-raising activities damage the forest. The existence of a market for rainforest products is, therefore, not sufficient on its own to prevent habitat destruction or the overexploitation of the resource and a more sophisticated approach to sustainability is required. Development of a market in ethically traded Brazil nuts might be one solution. PMID- 16701270 TI - Limpets break Dollo's Law. AB - A new molecular phylogeny of the limpet molluscs (Calyptraeidae) reveals that coiled shells have independently re-evolved at least once in this family, which is a violation of Dollo's Law that complex ancestral states, once lost, are never reacquired. Reacquisition of the coiled ancestral state is remarkable in that uncoiled shells have been the most recent ancestral state for 20 million-100 million years. Adult coiling might have re-evolved by the mechanism of prolonging the period during which genes for coiling are expressed in larvae. This and other developmental mechanisms could provide general routes for maintaining the potential to produce traits lost in distant ancestors. PMID- 16701271 TI - The PhyloCode: naming of biodiversity at a crossroads. PMID- 16701272 TI - Implementing the PhyloCode. PMID- 16701273 TI - Clarifying biotic homogenization. PMID- 16701275 TI - The need for evidence-based conservation. AB - Much of current conservation practice is based upon anecdote and myth rather than upon the systematic appraisal of the evidence, including experience of others who have tackled the same problem. We suggest that this is a major problem for conservationists and requires a rethinking of the manner in which conservation operates. There is an urgent need for mechanisms that review available information and make recommendations to practitioners. We suggest a format for web-based databases that could provide the required information in accessible form. PMID- 16701276 TI - Epialleles via DNA methylation: consequences for plant evolution. AB - In plants, naturally occurring methylation of genes can affect the level of gene expression. Variation among individuals in the degree of methylation of a gene, termed epialleles, produces novel phenotypes that are heritable across generations. To date, ecologically important genes with methylated epialleles have been found to affect floral shape, vegetative and seed pigmentation, pathogen resistance and development in plants. Currently, the extent to which epiallelic variation is an important common contributor to phenotypic variation in natural plant populations and its fitness consequences are not known. Because epiallele phenotypes can have identical underlying DNA sequences, response to selection on these phenotypes is likely to differ from expectations based on traditional models of microevolution. Research is needed to understand the role of epialleles in natural plant populations. Recent advances in molecular genetic techniques could enable population biologists to screen for epiallelic variants within plant populations and disentangle epigenetic from more standard genetic sources of phenotypic variance, such as additive genetic variance, dominance variance, epistasis and maternal genetic effects. PMID- 16701277 TI - The evolution of supertrees. AB - Supertrees result from combining many smaller, overlapping phylogenetic trees into a single, more comprehensive tree. As such, supertree construction is probably as old as the field of systematics itself, and remains our only way of visualizing the Tree of Life as a whole. Over the past decade, supertree construction has gained a more formal, objective footing, and has become an area of active theoretical and practical research. Here, I review the history of the supertree approach, focusing mainly on its current implementation. The supertrees of today represent some of the largest, complete phylogenies available for many groups, but are not without their critics. I conclude by arguing that the ever growing molecular revolution will result in supertree construction taking on a new role and implementation in the future for analyzing large DNA sequence matrices as part of a divide-and-conquer phylogenetic approach. PMID- 16701278 TI - Genic capture and resolving the lek paradox. AB - The genic capture hypothesis offers a resolution to the question of how genetic variation in male sexually selected traits is maintained in the face of strong female preferences. The hypothesis is that male display traits are costly to produce and hence depend upon overall condition, which itself is dependent upon genes at many loci. Few attempts have been made to test the assumptions and predictions of the genic capture hypothesis rigorously and, in particular, little attention has been paid to determining the genetic basis of condition. Such tests are crucial to our understanding of the maintenance of genetic variation and in the evaluation of recent models that propose a role for sexual selection in the maintenance of sex. Here, we review approaches to testing the link between genetically determined condition and levels of sexual trait expression and consider the probable importance of deleterious mutations. PMID- 16701279 TI - What is genetic quality? AB - Mate choice is favored by indirect selection if choosy females mate with males of high genetic quality. We believe, however, that testing hypotheses about indirect selection has been constrained by how we conceptualize and therefore empirically measure male genetic quality. Here, we argue that genetic quality is the breeding value of an individual for total fitness. We can therefore learn little about genetic quality from measures of only a few fitness components. We explain breeding value for total fitness, drawing on concepts from life-history theory and quantitative genetics, and suggest how approaches incorporating these insights might result in empirical progress. PMID- 16701280 TI - Biotelemetry: a mechanistic approach to ecology. AB - Remote measurement of the physiology, behaviour and energetic status of free living animals is made possible by a variety of techniques that we refer to collectively as 'biotelemetry'. This set of tools ranges from transmitters that send their signals to receivers up to a few kilometers away to those that send data to orbiting satellites and, more frequently, to devices that log data. They enable researchers to document, for long uninterrupted periods, how undisturbed organisms interact with each other and their environment in real time. In spite of advances enabling the monitoring of many physiological and behavioural variables across a range of taxa of various sizes, these devices have yet to be embraced widely by the ecological community. Our review suggests that this technology has immense potential for research in basic and applied animal ecology. Efforts to incorporate biotelemetry into broader ecological research programs should yield novel information that has been challenging to collect historically from free-ranging animals in their natural environments. Examples of research that would benefit from biotelemetry include the assessment of animal responses to different anthropogenic perturbations and the development of life time energy budgets. PMID- 16701281 TI - Professor John Maynard Smith 1920-2004. PMID- 16701282 TI - Superparasitism: a non-adaptive strategy? AB - Although once thought to be detrimental, superparasitism (where a host is parasitized more than once) by solitary parasitoids is now accepted to be an adaptive strategy. However, a recent study reveals that this might not always be the case. Varaldi et al. show that the superparasitism behaviour of the wasp Leptopilina boulardi is caused by a vertically and horizontally transmitted infectious agent. A reinterpretation of the adaptive significance of superparasitism in this species might therefore be required. PMID- 16701283 TI - Good news for sea turtles. AB - Following the overexploitation of sea turtle populations, conservation measures are now in place in many areas. However, the overall impact of these measures is often unknown because there are few long time-series showing trends in population sizes. In a recent paper, George Balazs and Milani Chaloupka chart the number of green turtles Chelonia mydas nesting in Hawaii over the past 30 years and reveal a remarkably quick increase in the size of this population following the instigation of conservation measures during the 1970s. Importantly, this work shows how even a small population of sea turtles can recover rapidly, suggesting that Allee effects do not impede conservation efforts in operation worldwide. PMID- 16701284 TI - Stochastic population theory faces reality in the laboratory. AB - Understanding the factors that affect most severely the extinction risk of populations is crucial for maintaining biodiversity. An important general pattern derived from stochastic population theory is that time to extinction should decrease with increasing environmental stochasticity. Drake and Lodge recently provided one of the first pieces of experimental support for this simple prediction by artificially manipulating the dynamics of populations of Daphnia. A future challenge will be to include both demographic stochasticity and environmental stochasticity in such studies. PMID- 16701285 TI - Are carotenoids a red herring in sexual display? PMID- 16701286 TI - Skeletons, noise and population growth: the end of an old debate? AB - Population dynamics models remain largely deterministic, although the presence of random fluctuations in nature is well recognized. This deterministic approach is based on the implicit assumption that systems can be separated into a deterministic part that captures the essential features of the system and a random part that can be neglected. But is it possible, in general, to understand population dynamics without the explicit consideration of random fluctuations? Here, we suggest perhaps not, and argue that the dynamics of many systems are a result of interactions between the deterministic nonlinear skeleton and noise. PMID- 16701287 TI - Modelling the past and future of whales and whaling. AB - Historical reconstruction of the population dynamics of whales before, during and after exploitation is crucial to marine ecological restoration and for the consideration of future commercial whaling. Population dynamic models used by the International Whaling Commission require historical catch records, estimates of intrinsic rates of increase and current abundance, all of which are subject to considerable uncertainty. Population genetic parameters can be used for independent estimates of historical demography, but also have large uncertainty, particularly for rates of mutational substitution and gene flow. At present, demographic and genetic estimates of pre-exploitation abundance differ by an order of magnitude and, consequently, suggest vastly different baselines for judging recovery. Here, we review these two approaches and suggest the need for a synthetic analytical framework to evaluate uncertainty in key parameters. Such a framework could have broad application to modelling both historical and contemporary population dynamics in other exploited species. PMID- 16701288 TI - Behavioral syndromes: an ecological and evolutionary overview. AB - Recent studies suggest that populations and species often exhibit behavioral syndromes; that is, suites of correlated behaviors across situations. An example is an aggression syndrome where some individuals are more aggressive, whereas others are less aggressive across a range of situations and contexts. The existence of behavioral syndromes focuses the attention of behavioral ecologists on limited (less than optimal) behavioral plasticity and behavioral carryovers across situations, rather than on optimal plasticity in each isolated situation. Behavioral syndromes can explain behaviors that appear strikingly non-adaptive in an isolated context (e.g. inappropriately high activity when predators are present, or excessive sexual cannibalism). Behavioral syndromes can also help to explain the maintenance of individual variation in behavioral types, a phenomenon that is ubiquitous, but often ignored. Recent studies suggest that the behavioral type of an individual, population or species can have important ecological and evolutionary implications, including major effects on species distributions, on the relative tendencies of species to be invasive or to respond well to environmental change, and on speciation rates. Although most studies of behavioral syndromes to date have focused on a few organisms, mainly in the laboratory, further work on other species, particularly in the field, should yield numerous new insights. PMID- 16701289 TI - Are natural microcosms useful model systems for ecology? AB - Several recent, high-impact ecological studies feature natural microcosms as tools for testing effects of fragmentation, metacommunity theory or links between biodiversity and ecosystem processes. These studies combine the microcosm advantages of small size, short generation times, contained structure and hierarchical spatial arrangement with advantages of field studies: natural environmental variance, 'openness' and realistic species combinations with shared evolutionary histories. This enables tests of theory pertaining to spatial and temporal dynamics, for example, the effects of neighboring communities on local diversity, or the effects of biodiversity on ecosystem function. Using examples, we comment on the position of natural microcosms in the roster of ecological research strategies and tools. We conclude that natural microcosms are as versatile as artificial microcosms, but as complex and biologically realistic as other natural systems. Research to date combined with inherent attributes of natural microcosms make them strong candidate model systems for ecology. PMID- 16701290 TI - Roles of parasites in animal invasions. AB - Biological invasions are global threats to biodiversity and parasites might play a role in determining invasion outcomes. Transmission of parasites from invading to native species can occur, aiding the invasion process, whilst the 'release' of invaders from parasites can also facilitate invasions. Parasites might also have indirect effects on the outcomes of invasions by mediating a range of competitive and predatory interactions among native and invading species. Although pathogen outbreaks can cause catastrophic species loss with knock-on effects for community structure, it is less clear what impact persistent, sub-lethal parasitism has on native-invader interactions and community structure. Here, we show that the influence of parasitism on the outcomes of animal invasions is more subtle and wide ranging than has been previously realized. PMID- 16701291 TI - The comparative method in conservation biology. AB - The phylogenetic comparative approach is a statistical method for analyzing correlations between traits across species. Whilst it has revolutionized evolutionary biology, can it work for conservation biology? Although it is correlative, advocates of the comparative method hope that it will reveal general mechanisms in conservation, provide shortcuts for prioritizing conservation research, and enable us to predict which species will experience (or create) problems in the future. Here, we ask whether these stated management goals are being achieved. We conclude that comparative methods are stimulating research into the ecological mechanisms underlying conservation, and are providing information for preemptive screening of problem species. But comparative analyses of extinction risk to date have tended to be too broad in scope to provide shortcuts to conserving particular endangered species. Correlates of vulnerability to conservation problems are often taxon, region and threat specific, so models must be narrowly focused to be of maximum practical use. PMID- 16701292 TI - The perils of payoff: corruption as a threat to global biodiversity. AB - Corruption is a worldwide phenomenon, particularly in many developing countries, which contain a large proportion of global biodiversity. Most alarming, from a biodiversity-conservation perspective, is the frequent corruption of government officials who manage valuable natural resources, such as timber, oil and precious minerals. A recent study by Joyotee Smith and colleagues describes rampant corruption in the timber industry of Indonesia, and shifts in the prevalence of different types of corruption as the country has become destabilized politically. By placing corruption into a conceptual framework, Smith et al. provide important insights into how developing nations and their natural resources can be besieged by corruption. PMID- 16701293 TI - Plant VOC emissions: making use of the unavoidable. AB - All plants emit substantial amounts of phytogenic volatile organic compounds (PVOCs), which include alkanes, alkenes, alcohols, aldehydes, eters, esters and carboxylic acids. Defence, communication and/or protection against extreme conditions have been proposed as reasons for these emissions. However, Rosenstiel and colleagues have recently proposed that emission of PVOCs represents a metabolic 'safety valve' by preventing the unnecessary sequestration of phosphates. Additionally, Niinemets and colleagues suggest that the emission rates of some PVOCs are determined by the principal physicochemical characteristics of the emitted compounds, such as their solubility, volatility and diffusivity, rather than by physiological mechanisms, such as their synthesis rates. These two new studies lead to the hypothesis that there is not necessarily a specific role for every PVOC emitted, given that their emission is unavoidable as result of their volatility. However, in some cases, natural selection has worked to take advantage of this volatility. PMID- 16701294 TI - Hybridization, rare alleles and adaptive radiation. PMID- 16701296 TI - Monkeys, mirrors, mark tests and minds. PMID- 16701298 TI - The effect of natal experience on habitat preferences. AB - Several important problems in ecology, evolution and conservation biology are affected by habitat selection in dispersing animals. Experience in the natal habitat has long been considered a potential source of variation in the habitat preferences displayed when dispersers select a post-dispersal habitat. However, the taxonomic breadth of this phenomenon is underappreciated, in part because partially overlapping, taxon-specific definitions in the literature have discouraged communication. Here, we explore the phenomenon of natal habitat preference induction (NHPI) and demonstrate that NHPI has been observed in a broad range of animal taxa. We consider the potential adaptive significance of NHPI, identify implications of its occurrence for problems in evolution, ecology and conservation biology, and encourage further study of this phenomenon. PMID- 16701299 TI - Evolution in invasive plants: implications for biological control. AB - Evidence is increasing that invasive plants can undergo rapid adaptive evolution during the process of range expansion. Here, we argue that evolutionary change during invasions will also affect plant-antagonist interactions and, thus, will have important implications for biological control programmes targeted at invasive plants. We explore how altered selection in the new range might influence the evolution of plant defence (resistance and tolerance) and life history. The degree to which such evolutionary processes might affect biological control efficacy is largely unexplored. We hope that, by testing the hypotheses that we propose here, a closer link can be established between biological control and evolutionary biology, to the benefit of both disciplines. PMID- 16701300 TI - Evo-devo perspectives on segmentation: model organisms, and beyond. AB - Bilaterian animals show a diverse array of segmental patterns and segmentation processes. Differences in pattern and process emerge both in comparisons of taxa and among sets of serial structures within one animal. Diversity in developmental mechanisms of segmentation and their genetic control is reflected in the modes in which segmentation evolves, which are difficult to accommodate within the traditional concept of segments as modular building blocks. Thus, in spite of the apparent simplicity of segmental patterns, studying the evolution of segmentation requires an approach that, in an adequate comparative framework, combines the efforts of researchers of genes, cells, embryos and post-embryonic stages. PMID- 16701301 TI - Molecules consolidate the placental mammal tree. AB - Deciphering relationships among the orders of placental mammals remains an important problem in evolutionary biology and has implications for understanding patterns of morphological character evolution, reconstructing the ancestral placental genome, and evaluating the role of plate tectonics and dispersal in the biogeographic history of this group. Until recently, both molecular and morphological studies provided only a limited and questionable resolution of placental relationships. Studies based on larger and more diverse molecular datasets, and using an array of methodological approaches, are now converging on a stable tree topology with four major groups of placental mammals. The emerging tree has revealed numerous instances of convergent evolution and suggests a role for plate tectonics in the early evolutionary history of placental mammals. The reconstruction of mammalian phylogeny illustrates both the pitfalls and the powers of molecular systematics. PMID- 16701302 TI - Genomics for the ecological toolbox. AB - Genomics technologies have expanded the types of question that can be addressed in human genetics and health and in fields such as ecology. Genome-scale approaches provide ways to examine physiological changes that occur when a pathogen invades a host, the response of an organism to a change in its environment, and the way in which changes in the microbial community affect ecosystem function. Here, we examine successful applications of genomics to ecological science to date and describe ways that classic ecological research disciplines might benefit from genomic approaches. We also address some of the challenges of using this methodology, and discuss how ecological researchers embracing these approaches enhance its effectiveness in applications such as gene hunting and gene expression analysis. PMID- 16701303 TI - Tree diseases and landscape processes: the challenge of landscape pathology. AB - Forest pathology inherently involves a landscape perspective, because tree pathogens propagate according to heterogeneous spatial patterns of flow and isolation. Landscape pathology is a field that is now emerging from the transdisciplinary cooperation of forest pathologists with landscape ecologists. Here, we review recent broad-scale assessments of tree disease risk, investigations of site and host preferences for several root rot pathogens, and regional historical analyses of pathogen outbreak in plantations. Crucial topics include fragmentation effects on pathogen spread and geophysical features that predispose forest patches to disease expression. Recent methodological developments facilitate the spatially explicit analysis of reciprocal coarse scale relationships among hosts and pathogens. Landscape pathology studies fill a significant research gap in the context of our understanding of sustainable forest management, the introduction of exotic organisms and how climate change might affect the spread of disease. PMID- 16701304 TI - Many wrongs: the advantage of group navigation. AB - Research into the puzzling phenomena of animal navigation and aggregation has proceeded along two distinct lines. Study of navigation generally focuses on the orientation ability of the individual without reference to the implications of group membership. A simple principle (the 'many wrongs principle'), first proposed by Bergman and Donner in 1964, and developed by both Hamilton and Wallraff three decades ago, provides a link between these lines of current interest by suggesting that navigational accuracy increases with group size. With unprecedented scope for testing the hypotheses it generates, it is now time that the many wrongs principle is exhumed. PMID- 16701305 TI - Evolution in parallel: new insights from a classic system. AB - Neo-darwinists have long argued that parallel evolution, the repeated evolution of similar phenotypes in closely related lineages, is caused by the action of similar environments on alleles at many loci of small effect. A more controversial possibility is that the genetic architecture of traits initiates parallelism, sometimes through fixation of alleles of large effect. Recent research (by Cole et al., Colosimo et al., Cresko et al., and Shapiro et al.) offers the surprising insight that reduction in two armor traits of threespine stickleback is governed by independently segregating major loci as well as additional quantitative trait loci (QTL), and that alleles at the same major loci are associated with parallel phenotypes in globally distributed populations. This research suggests the emergence of a new and exciting vertebrate model system for evolutionary genetics. PMID- 16701306 TI - Pesticides, mortality and population growth rate. PMID- 16701308 TI - Taxonomic inflation: its influence on macroecology and conservation. AB - Species numbers are increasing rapidly. This is due mostly to taxonomic inflation, where known subspecies are raised to species as a result in a change in species concept, rather than to new discoveries. Yet macroecologists and conservation biologists depend heavily on species lists, treating them as accurate and stable measures of biodiversity. Deciding on a standardized, universal species list might ameliorate the mismatch between taxonomy and the uses to which it is put. However, taxonomic uncertainty is ultimately due to the evolutionary nature of species, and is unlikely to be solved completely by standardization. For the moment, at least, users must acknowledge the limitations of taxonomic species and avoid unrealistic expectations of species lists. PMID- 16701309 TI - Are invasive species a major cause of extinctions? AB - The link between species invasions and the extinction of natives is widely accepted by scientists as well as conservationists, but available data supporting invasion as a cause of extinctions are, in many cases, anecdotal, speculative and based upon limited observation. We pose the question, are aliens generally responsible for widespread extinctions? Our goal is to prompt a more critical synthesis and evaluation of the available data, and to suggest ways to take a more scientific, evidence-based approach to understanding the impact of invasive species on extinctions. Greater clarity in our understanding of these patterns will help us to focus on the most effective ways to reduce or mitigate extinction threats from invasive species. PMID- 16701310 TI - Bayesian inference of character evolution. AB - Much recent progress in evolutionary biology is based on the inference of ancestral states and past transformations in important traits on phylogenetic trees. These exercises often assume that the tree is known without error and that ancestral states and character change can be mapped onto it exactly. In reality, there is often considerable uncertainty about both the tree and the character mapping. Recently introduced Bayesian statistical methods enable the study of character evolution while simultaneously accounting for both phylogenetic and mapping uncertainty, adding much needed credibility to the reconstruction of evolutionary history. PMID- 16701311 TI - Chromosomal inversion polymorphisms and adaptation. AB - Chromosomal inversion polymorphisms continue to be identified from an increasing number of populations of insects, plants, bacteria and humans. In the fruit fly Drosophila, chromosomal polymorphisms were used in classic studies of natural selection. Recent molecular genetic studies suggest that inversion polymorphisms are dynamical systems. These studies also indicate patterns of disequilibrium and variation that are consistent with co-adapted gene complexes. Although these complexes have yet to be identified, recent studies have identified traits, such as body size, that are linked to inversion polymorphisms. Selection acting on these polymorphisms is strong because latitudinal clines in inversion frequency become re-established rapidly after a new continent is colonized. A combined molecular and phenotypic approach is helping to identify the role of inversion polymorphisms in adaptive divergence, but the genes responsible for associations between traits and inversion polymorphisms remain to be identified. PMID- 16701312 TI - The alluring simplicity and complex reality of genetic rescue. AB - A series of important new theoretical, experimental and observational studies demonstrate that just a few immigrants can have positive immediate impacts on the evolutionary trajectory of local populations. In many cases, a low level of immigration into small populations has produced fitness benefits that are greater than those predicted by theoretical models, resulting in what has been termed 'genetic rescue'. However, the opposite result (reduced fitness) can also be associated with immigration of genetically divergent individuals. Central to our understanding of genetic rescue are complex interactions among fundamental concepts in evolutionary and population biology, including both genetic and non genetic (environmental, behavioral and demographic) factors. Developing testable models to predict when genetic rescue is likely to occur is a daunting challenge that will require carefully controlled, multi-generation experiments as well as creative use of information from natural 'experiments'. PMID- 16701313 TI - New developments in museum-based informatics and applications in biodiversity analysis. AB - Information from natural history collections (NHCs) about the diversity, taxonomy and historical distributions of species worldwide is becoming increasingly available over the Internet. In light of this relatively new and rapidly increasing resource, we critically review its utility and limitations for addressing a diverse array of applications. When integrated with spatial environmental data, NHC data can be used to study a broad range of topics, from aspects of ecological and evolutionary theory, to applications in conservation, agriculture and human health. There are challenges inherent to using NHC data, such as taxonomic inaccuracies and biases in the spatial coverage of data, which require consideration. Promising research frontiers include the integration of NHC data with information from comparative genomics and phylogenetics, and stronger connections between the environmental analysis of NHC data and experimental and field-based tests of hypotheses. PMID- 16701314 TI - Is Her Majesty at home? AB - When Queen Elizabeth is at home in Buckingham Palace, tradition has it that the Royal Standard is raised, so that all may know the fact. Although it is not crucial for most of us to know whether Her Majesty is home, it is in social insects. Endler et al. have recently shown how an ant queen signals her presence to her remote workers: she marks her eggs. This is significant because it provides insight into how queens maintain reproductive monopoly within their colonies. PMID- 16701315 TI - Lepidopteran microsatellite DNA: redundant but promising. AB - The development of microsatellite DNA markers for use with butterflies and moths is extremely difficult for no apparent reason. New findings demonstrate that many lepidopteran microsatellite sequences exist in multiple copies in the genome, and have similar or almost identical flanking regions. These findings provide a compelling explanation for the low efficiency of microsatellite isolation in Lepidoptera, and might also shed light on the evolutionary dynamics of microsatellite sequences in these insects and other organisms. PMID- 16701316 TI - The rise of birds and mammals: are microevolutionary processes sufficient for macroevolution? AB - It is a basis of darwinian evolution that the microevolutionary mechanisms that can be studied in the present are sufficient to account for macroevolution. However, this idea needs to be tested explicitly, as highlighted here by the example of the superceding of dinosaurs and pterosaurs by birds and placental mammals that occurred near the Cretaceous/Tertiary boundary approximately 65 million years ago. A major problem for testing the sufficiency of microevolutionary processes is that independent ideas (such as the existence of an extraterrestrial impact, and the extinction of dinosaurs) were linked without the evidence for each idea being evaluated separately. Here, we suggest and discuss five testable models for the times and divergences of modern mammals and birds. Determination of the model that best represents these events will enable the role of microevolutionary mechanisms to be evaluated. The question of the sufficiency of microevolutionary processes for macroevolution is solvable, and available evidence supports an important role for biological processes in the initial decline of dinosaurs and pterosaurs. PMID- 16701317 TI - A role for immunology in invasion biology. AB - Invasive species are of increasing conservation and economic concern, yet mechanisms underlying invasions remain poorly understood. We propose that variation in immune defences might help explain why only some introduced populations become invasive. Introduced species escape many of their native diseases, but also face novel pathogens that can induce costly, and sometimes deadly, immune responses in naive hosts. Therefore, favouring less resource demanding and dangerous defence mechanisms and allocating a greater proportion of resources to growth and reproduction should favour invasion. Specifically, we argue that successful invaders should reduce costly systemic inflammatory responses, which are associated with fever and metabolic and behavioural changes, and rely more heavily on less expensive antibody-mediated immunity. Here we provide supporting arguments for this hypothesis and generate predictions that are testable using tools from the growing field of ecological immunology. PMID- 16701318 TI - Simulation as experiment: a philosophical reassessment for biological modeling. AB - Some scientific modelers suggest that complex simulation models that mimic biological processes should have a limited place in ecological and evolutionary studies. However, complex simulation models can have a role that is different from that of simpler models that are designed to be fit to data. Simulation can be viewed as another kind of experimental system and should be analyzed as such. Here, I argue that current discussions in the philosophy of science and in the physical sciences fields about the use of simulation as an experimental system have important implications for biology, especially complex sciences such as evolution and ecology. Simulation models can be used to mimic complex systems, but unlike nature, can be manipulated in ways that would be impossible, too costly or unethical to do in natural systems. Simulation can add to theory development and testing, can offer hypotheses about the way the world works and can give guidance as to which data are most important to gather experimentally. PMID- 16701319 TI - Emerging infectious diseases of plants: pathogen pollution, climate change and agrotechnology drivers. AB - Emerging infectious diseases (EIDs) pose threats to conservation and public health. Here, we apply the definition of EIDs used in the medical and veterinary fields to botany and highlight a series of emerging plant diseases. We include EIDs of cultivated and wild plants, some of which are of significant conservation concern. The underlying cause of most plant EIDs is the anthropogenic introduction of parasites, although severe weather events are also important drivers of disease emergence. Much is known about crop plant EIDs, but there is little information about wild-plant EIDs, suggesting that their impact on conservation is underestimated. We conclude with recommendations for improving strategies for the surveillance and control of plant EIDs. PMID- 16701320 TI - SINEs of speciation: tracking lineages with retroposons. AB - The value of short interspersed elements (SINEs) for diagnosing common ancestry is being expanded to examine the differential sorting of lineages through the course of speciation events. Because most SINEs are neutral markers of identical descent, are not precisely excised from the genome and have a known ancestral condition, they are advantageous for reconciling gene trees and species trees with minimal phylogenetic error. A population perspective on SINE evolution combined with coalescence theory provides a context for investigating the phenomenon of ancestral polymorphism and its role in producing incongruent SINE insertion patterns among multiple loci. Studies of human Alu repeats demonstrate the value of young polymorphic SINEs for assessing human genomic diversity and tracking ancient demographics of human populations, whereas incongruent insertion patterns revealed by older fixed SINE loci, such as those in African cichlid fishes, contain information that might help identify ancient radiations that are otherwise obscured by accumulated mutations in sequence data. Here, we review the utility of retroposons for inferring common ancestry, discuss limits to the method, and clarify confusion by providing examples from the literature that illustrate how discordant multi-locus insertion patterns of retroelements can indicate lineage-sorting events that should not be misinterpreted as phylogenetic noise. PMID- 16701321 TI - Choosing mates: good genes versus genes that are a good fit. AB - Female choice for male ornamental traits is widely accepted as a mechanism by which females maximize their reproductive success and/or offspring quality. However, there is an increasing empirical literature that shows a fitness benefit of genetic diversity and a tendency for females to use genetic dissimilarity as a criterion for mate choice. This genetic compatibility hypothesis for female mate choice presents a paradox. How can females use both an absolute criterion, such as male ornamentation, and a relative criterion, such as genetic dissimilarity, to choose their mates? Here, we present potential solutions for this dilemma and the empirical evidence supporting them. The interplay between these two contrasting forms of female mate choice presents an exciting empirical and theoretical challenge for evolutionary ecologists. PMID- 16701322 TI - Measuring inbreeding depression in the wild: the old ways are the best. AB - Measuring inbreeding depression in normally outbreeding natural populations is an uphill task, because it requires inbreeding coefficients that are calculated from pedigrees. Instead of calculating inbreeding coefficients directly, several studies have reported the use of microsatellite-derived metrics, such as heterozygosity, to infer relative inbreeding among individuals. In two new papers, Slate et al. and Balloux et al. show that these molecular metrics are often only weakly correlated with inbreeding coefficients, and that correlations between heterozygosity and fitness require a new interpretation. PMID- 16701323 TI - Red leaves, insects and coevolution: a red herring? AB - W.D. (Bill) Hamilton proposed that coevolution between plants and herbivorous insects explains the bright autumnal colouration of leaves. Accordingly, plants invest in bright signals to reduce their herbivore load, whereas insects use these bright signals to identify less-defended hosts more efficiently. Archetti and Brown have recently revisited this theory by explaining its basic predictions and providing new research perspectives. Their work presents an important basis to our understanding of non-green leaf colouration, provided that alternative adaptive explanations on the photoprotective and antioxidant role of leaf pigments, or their possible function in crypsis to herbivores are incorporated into future research. PMID- 16701324 TI - Vicious circles: positive feedback in major evolutionary and ecological transitions. AB - Evolutionary biologists and ecologists often focus on equilibrium states that are subject to forms of negative feedback, such as optima for phenotypic traits or regulation of population sizes. However, recent theoretical and empirical studies show how positive feedback can be instrumental in driving many of the most important and spectacular processes in evolutionary ecology, including the evolution of sex and genetic systems, mating systems, life histories, complex cooperation in insects and humans, ecological specialization, species diversity, species ranges, speciation and extinction. Taken together, this work suggests that positive feedback is more common than is generally appreciated, and that its self-reinforcing dynamics generate the conditions for changes that might otherwise be difficult or impossible for selection or other mechanisms to achieve. Testing for positive feedback requires analysing each causal link in feedback loops, tracking genetic, character and population-dynamic changes across generations, and elucidating the conditions that can result in self-reinforcing change. PMID- 16701325 TI - Tropical biologists, local people and conservation: new opportunities for collaboration. AB - Tropical biologists need help. Examples show that local people can be trained to be effective parataxonomists, greatly assisting efforts to document and assess tropical biodiversity. Local collaborations also offer promising ways with which to improve natural resource management and conservation. However, for several reasons, most biologists remain slow to approve and implement these approaches. The challenges and potentials need evaluation and neglect means that opportunities are being missed. Here, we consider experiences of local collaborations and discuss obstacles to their wider implementation. We urge tropical biologists to recognize and embrace the opportunities provided by working with local people. PMID- 16701326 TI - Historical biogeography, ecology and species richness. AB - Ecology and historical (phylogeny-based) biogeography have much to offer one another, but exchanges between these fields have been limited. Historical biogeography has become narrowly focused on using phylogenies to discover the history of geological connections among regions. Conversely, ecologists often ignore historical biogeography, even when its input can be crucial. Both historical biogeographers and ecologists have more-or-less abandoned attempts to understand the processes that determine the large-scale distribution of clades. Here, we describe the chasm that has developed between ecology and historical biogeography, some of the important questions that have fallen into it and how it might be bridged. To illustrate the benefits of an integrated approach, we expand on a model that can help explain the latitudinal gradient of species richness. PMID- 16701327 TI - Cytonuclear coevolution: the genomics of cooperation. AB - Without mitochondria we would be in big trouble, and there would be a global biological energy crisis if it were not for chloroplasts. Fortunately, genomic evolution over the past two billion years has ensured that the functions of these key organelles are with us to stay. Whole-genome analyses have not only proven that mitochondria and chloroplasts are descended from formerly free-living bacteria, but have also shown that it is difficult to define eukaryotes without reference to the fusion and coevolution of host and endosymbiont genomes. Here, we review how the macro- and microevolutionary insights that follow from the genomics of cytonuclear interactions are uniting molecular evolution, structural proteomics, population genetics and problems in aging and disease. Our goals are to clarify the coevolutionary events that have governed nuclear and organelle evolution, and to encourage further critical analyses of these interactions as problems in the study of co-adapted gene complexes. PMID- 16701328 TI - Southeast Asian biodiversity: an impending disaster. AB - Southeast Asia has the highest relative rate of deforestation of any major tropical region, and could lose three quarters of its original forests by 2100 and up to 42% of its biodiversity. Here, we report on the current state of its biota and highlight the primary drivers of the threat of extinction now faced by much of the unique and rich fauna and flora of the region. Furthermore, the known impacts on the biodiversity of Southeast Asia are likely to be just the tip of the iceberg, owing to the paucity of research data. The looming Southeast Asian biodiversity disaster demands immediate and definitive actions, yet such measures continue to be constrained by socioeconomic factors, including poverty and lack of infrastructure. Any realistic solution will need to involve a multidisciplinary strategy, including political, socioeconomic and scientific input, in which all major stakeholders (government, non-government, national and international organizations) must participate. PMID- 16701329 TI - Charophyte algae and land plant origins. AB - The charophyte algae are six distinct groups of mostly freshwater green algae that are related to modern land plants. Charophyte algae exhibit diverse morphologies and reproductive strategies, from unicells to branching erect forms, and from swimming asexual spores to sex involving eggs and sperm, respectively. The green algae known as stoneworts (Charales) are suggested to be the extant sister group to all land plants, although the phylogeny is not conclusive. Here we review recent molecular phylogenetic work on the charophyte algae and its implications for our understanding of the origins of land plants and of characters in their aquatic ancestors that might have played a role in the explosive diversification of plants on land. PMID- 16701330 TI - How isolated is Antarctica? AB - The traditional view of Antarctica and the surrounding Southern Ocean as an isolated system is now being challenged by the recent discovery at the Antarctic Peninsula of adult spider crabs Hyas areneus from the North Atlantic and of larvae of subpolar marine invertebrates. These observations question whether the well described biogeographical similarities between the benthic fauna of the Antarctic Peninsula and the Magellan region of South America result from history (the two regions were once contiguous), or from a previously unrecognized low level of faunal exchange. Such exchange might be influenced by regional climate change, and also exacerbated by changes in human impact. PMID- 16701331 TI - The evolution of large size: how does Cope's Rule work? AB - Cope's Rule is the tendency for organisms in evolving lineages to increase in size over time. The concept is detailed in many textbooks, but has rarely been demonstrated. Many suggestions of the benefits of large body size exist, but none has yet been confirmed empirically. Using a large-scale analysis of recent studies, Kingsolver and Pfennig have now shown how size benefits survival, mating success and fecundity, and they provide convincing arguments for a mechanism that is capable of driving Cope's Rule. PMID- 16701332 TI - Taxonomic inflation: species concept or historical geopolitical bias? PMID- 16701333 TI - Taxonomic inflation, species concepts and global species lists. PMID- 16701335 TI - The hippocampus, spatial memory and food hoarding: a puzzle revisited. AB - Behavioural ecology assumes that cognitive traits and their underlying neural substrates are shaped by natural selection in much the same way as morphological traits are, resulting in adaptation to the natural environment of the species concerned. Recently, however, the 'neuroecology' approach of attempting to gain insight into brain structure and function by testing predictions about variation in brain structure based on knowledge of the lifestyle of the animal has been criticized on the grounds that such an adaptationist view cannot provide insight into the underlying mechanisms. Furthermore, the criticism has focussed on attempts to use variation in demand for spatial memory and in hippocampal size as a basis for predicting variation in cognitive abilities. Here, we revisit this critique against the field of so-called 'neuroecology' and argue that using knowledge of the natural history of animals has lead to a better understanding of the interspecific variation in spatial abilities and hippocampal size, and to the generation of novel hypotheses and predictions. PMID- 16701336 TI - Organisms in nature as a central focus for biology. AB - Theories summarize science, tell us what to measure when we test hypotheses, and help us study nature better. Nevertheless, organisms themselves embody genetics, development, morphology, physiology and behavior, and they are the units of populations, communities and ecosystems. Biologists seek to understand organisms, their diversification and environmental relationships--not theories and experiments per se--and discoveries of new organisms and new facts about organisms reset the research cycles of hypothesis testing that underlie conceptually progressive science. I argue here that recent disagreements about the fate of natural history are thus more apparent than real and should not distract us from addressing important issues. The conservation of biodiversity requires factual knowledge of particular organisms, yet we know little or nothing about most species, and organismal diversity is often poorly represented in biological education. Accordingly, I urge those who are especially concerned with teaching and conservation to seek increased financial and curricular support for descriptive natural history, which is so fundamental to many of the applied facets of biology. PMID- 16701337 TI - What determines the citation frequency of ecological papers? AB - Citation frequencies of scientific articles are increasingly used for academic evaluation in various disciplines, including ecology. However, the factors affecting citation rates have not been extensively studied. Here, we examine the association between the citation frequency of ecological articles and various characteristics of journals, articles and authors. Our analysis shows that the annual citation rates of ecological papers are affected by the direction of the study outcome with respect to the hypothesis tested (supportive versus unsupportive evidence), by article length, by the number of authors, and by their country and university of affiliation. These results cast doubt on the validity of using citation counts as an objective and unbiased tool for academic evaluation in ecology. PMID- 16701338 TI - Conflict between parents over care. AB - Conflict between parents over care of young arises when the young benefit from the effort of both parents, but each parent suffers a reduction in future reproductive success as a consequence of its own effort. Here, we review existing models and argue that they fail to capture many important components of parental conflict. For example, we lack adequate models of how a parent should compensate for a reduction in the effort of its mate. These models should incorporate the process by which decisions are reached. Recent theory suggests that a parent benefits by handicapping itself, and more experimental and theoretical work on this topic could be fruitful. We also need more theoretical work on attractiveness that incorporates consistent interactions between males and females. PMID- 16701339 TI - Germs, genomes and genealogies. AB - Genetic diversity in pathogen species contains information about evolutionary and epidemiological processes, including the origins and history of disease, the nature of the selective forces acting on pathogen genes and the role of recombination in generating genetic novelty. Here, we review recent developments in these fields and compare the use of population genetic, or population-model based, approaches to phylogenetic, or population-model free, methodologies. We show how simple epidemiological models can be related to the ancestral, or coalescent, process underlying samples from pathogen species, enabling detailed inference about pathogen biology from patterns of molecular variation. PMID- 16701341 TI - A suitable job for a woman. PMID- 16701340 TI - Sperm in competition: not playing by the numbers. AB - The outcome of sperm competition is mediated largely by the relative numbers of sperm from competing males. However, substantial variation in features of sperm morphology and behaviour, such as length, longevity and motility, exists and researchers have suggested that this variation functions in postcopulatory sexual selection. Recent studies have determined the effect of these sperm-quality traits on fertilization success and a synthesis of this literature reveals that they are important in both sperm competition and cryptic female choice. To understand how postcopulatory sexual selection influences sperm traits, future research should determine sex-specific interactions that influence paternity, identify genetic correlations between ejaculate characters, quantify the relative costs of producing different sperm traits, and test assumptions of models of sperm quality evolution. Such research will shed light on what evolutionary pressures are responsible for the diversity in sperm morphometry and behaviour. PMID- 16701342 TI - Glacial cycles and the tempo of avian speciation. AB - How old are most species of North American birds? A longstanding debate centers on whether Pleistocene events caused substantial avian diversification. Two new syntheses of speciation times by Johnson and Cicero, and Weir and Schluter provide compelling evidence of recent Pleistocene speciation, whereas diversification rate analyses by Zink et al. suggest that most speciation events occurred earlier. Although these results are seemingly contradictory, when considered together they provide a robust and complementary perspective on the tempo of avian diversification in North America. PMID- 16701343 TI - Do some corals like it hot? AB - Global increases in sea temperatures threaten coral reef resilience because thermal stress can cause corals to bleach; that is, to lose their photosynthetic microalgal symbionts. Recent evidence suggests that some corals associate with genotypes of microalgae that resist future thermal stress, however, these genotypes might provide less energy for growth when thermal stresses are curtailed. Coral reef resilience depends on whether phenotypic and genotypic changes in host-symbiont associations can match projected increases in the frequency and severity of thermal stress, as well as on our ability to ameliorate continuing human impacts. PMID- 16701344 TI - Choosing mates: complementary versus compatible genes. PMID- 16701345 TI - The resurrection of oceanic dispersal in historical biogeography. AB - Geographical distributions of terrestrial or freshwater taxa that are broken up by oceans can be explained by either oceanic dispersal or vicariance in the form of fragmentation of a previously contiguous landmass. The validation of plate tectonics theory provided a global vicariance mechanism and, along with cladistic arguments for the primacy of vicariance, helped create a view of oceanic dispersal as a rare phenomenon and an explanation of last resort. Here, I describe recent work that suggests that the importance of oceanic dispersal has been strongly underestimated. In particular, molecular dating of lineage divergences favors oceanic dispersal over tectonic vicariance as an explanation for disjunct distributions in a wide variety of taxa, from frogs to beetles to baobab trees. Other evidence, such as substantial gene flow among island populations of Anolis lizards, also indicates unexpectedly high frequencies of oceanic dispersal. The resurrection of oceanic dispersal is the most striking aspect of a major shift in historical biogeography toward a more even balance between vicariance and dispersal explanations. This new view implies that biotas are more dynamic and have more recent origins than had been thought previously. A high frequency of dispersal also suggests that a fundamental methodological assumption of many biogeographical studies--that vicariance is a priori a more probable explanation than dispersal--needs to be re-evaluated and perhaps discarded. PMID- 16701346 TI - Critical science gaps impede use of no-take fishery reserves. AB - As well as serving valuable biodiversity conservation roles, functioning no-take fishery reserves protect a portion of the fishery stock as insurance against future over-fishing. So long as there is adequate compliance by the fishing community, it is likely that they will also sustain and even enhance fishery yields in the surrounding area. However, there are significant gaps in scientific knowledge that must be filled if no-take reserves are to be used effectively as fishery management tools. Unfortunately, these gaps are being glossed over by some uncritical advocacy. Here, we review the science, identify the most crucial gaps, and suggest ways to fill them, so that a promising management tool can help meet the growing challenges faced by coastal marine fisheries. PMID- 16701347 TI - Towards an evolutionary ecology of life in soil. AB - The soil-microbe system is one of the most diverse components of the terrestrial ecosystem. The origin of this diversity, and its relation to the life-sustaining processes that are mediated by the resident microbial community, is still poorly understood. The inherent complexities necessitate a theoretical framework that integrates ecological and evolutionary approaches and which embraces the physical heterogeneity of the soil environment. Such a framework is currently lacking, although recent advances in theory and experimentation are beginning to identify the essential ingredients. Here, we review and evaluate the relevance of current modelling approaches, and propose a new synthesis of an evolutionary ecology of life in soil. Key elements include an account of dispersal, horizontal gene transfer, and the consideration of the physical and biological components of soil as an integrated complex adaptive system. PMID- 16701348 TI - Criticality and disturbance in spatial ecological systems. AB - Classical criticality describes sudden changes in the state of a system when underlying processes change slightly. At this transition, patchiness develops which lacks a characteristic or dominant spatial scale. Thus, criticality lies at the interface of two important subjects in ecology, threshold behavior and patchiness. Most ecological examples of criticality involve processes of disturbance and recovery; the spatial and temporal scales of these processes enable three different types of critical system to be distinguished: classical phase transitions, self organized criticality (SOC) and 'robust' criticality. Here, we review the properties defining these three types and their implications for threshold behavior and large intermittent temporal fluctuations, with examples taken from spatial stochastic models for predator-prey, infected susceptible, and disturbance-recovery interactions. In critical systems, spatial properties of patchiness alone are insufficient indicators of impending sudden changes, unless complemented by the spatial and temporal scales of disturbance and recovery themselves. PMID- 16701349 TI - Candidate genes for behavioural ecology. AB - In spite of millions of years of evolutionary divergence, the conservation of gene function is common across distant lineages. As such, genes that are known to influence behaviour in one organism are likely to influence similar behaviours in other organisms. Recent studies of the evolution of behaviour and morphological adaptation support this notion. Thus, the candidate gene approach offers great potential to expand our understanding of behavioural ecology. Changes in the expression of candidate genes can reveal their contribution to behavioural variation and/or phenotypic plasticity. Knowledge of gene function also enables experimental manipulation of behaviour in the lab and in the field. The candidate gene approach provides an accessible and useful tool for generating insights about animals that are not typically associated with genetic experimentation. PMID- 16701350 TI - New methods ring changes for the tree of life. AB - Relationships among prokaryotes and the origin of eukaryotes have both proven controversial, with results depending upon the gene sequences and methods used. Extensive horizontal gene transfer is one possible reason why inferring such deep phylogenetic relationships is difficult. In two recent papers, Lake and Rivera introduce new methods that can be used to reconstruct the genomic tree in the presence of horizontal gene transfers, but which suggest that a ring rather than a tree is a better representation of some parts of the history of life on Earth. PMID- 16701351 TI - Providing baselines for biodiversity measurement. PMID- 16701352 TI - 'Genetic restoration:' a more comprehensive perspective than 'genetic rescue'. PMID- 16701353 TI - Invasive species are a leading cause of animal extinctions. PMID- 16701354 TI - A phylogenetic approach to cultural evolution. AB - There has been a rapid increase in the use of phylogenetic methods to study the evolution of languages and culture. Languages fit a tree model of evolution well, at least in their basic vocabulary, challenging the view that blending, or admixture among neighbouring groups, was predominant in cultural history. Here, we argue that we can use language trees to test hypotheses about not only cultural history and diversification, but also bio-cultural adaptation. Phylogenetic comparative methods take account of the non-independence of cultures (Galton's problem), which can cause spurious statistical associations in comparative analyses. Advances in phylogenetic methods offer new possibilities for the analysis of cultural evolution, including estimating the rate of evolution and the direction of coevolutionary change of traits on the tree. They also enable phylogenetic uncertainty to be incorporated into the analyses, so that one does not have to treat phylogenetic trees as if they were known without error. PMID- 16701355 TI - Do early branching lineages signify ancestral traits? AB - A reverence for ancestors that has pre-occupied humans since time immemorial persists to the present. Reconstructing ancestry is the focus of many biological studies but failure to distinguish between present-day descendants and long-dead ancestors has led to incorrect interpretation of phylogenetic trees. This has resulted in erroneous reconstruction of traits such as morphology and ancestral areas. Misinterpretation becomes evident when authors use the terms 'basal' or 'early diverging' to refer to extant taxa. Here, we discuss the correct interpretation of trees and methods for reconstructing the ancestral features of organisms using recently developed statistical models. These models can be inaccurate unless they use information that is independent of phylogenies, such as genetics, molecular and developmental biology, functional morphology, geological and climatic processes, and the fossil record. PMID- 16701356 TI - Integrating ecology and developmental biology to explain the timing of frog metamorphosis. AB - Amphibian metamorphosis has long intrigued ecologists and developmental biologists, yet the two research programs have progressed separately and toward different goals. Plasticity in metamorphic timing has profound effects on fitness, which has prompted ecologists to develop and test models for predicting how environmental factors affect the size and age of metamorphosis. These models rely upon untested assumptions about the mechanisms for regulating growth and development. Whereas developmental biologists explicitly investigate these mechanisms at the hormonal and genetic levels, they largely ignore the role of environmental input. Recent developments in our understanding of the molecular biology of frog metamorphosis are revealing how these two research programs could be integrated. Here, I review these developments to test ecologists' assumptions about frog metamorphosis, and to present strategies for both research fields to investigate the mechanistic basis of metamorphic plasticity. PMID- 16701357 TI - Assignment methods: matching biological questions with appropriate techniques. AB - Assignment methods, which use genetic information to ascertain population membership of individuals or groups of individuals, have been used in recent years to study a wide range of evolutionary and ecological processes. In applied studies, the first step of articulating the biological question(s) to be addressed should be followed by selection of the method(s) best suited for the analysis. However, this first step often receives less attention than it should, and the recent proliferation of assignment methods has made the selection step challenging. Here, we review assignment methods and discuss how to match the appropriate methods with the underlying biological questions for several common problems in ecology and conservation (assessing population structure; measuring dispersal and hybridization; and forensics and mixture analysis). We also identify several topics for future research that should ensure that this field remains dynamic and productive. PMID- 16701358 TI - Functional aspects of song learning in songbirds. AB - The oscine passerines, or 'songbirds', are one of the few animal taxa in which individuals learn their vocal signals. Recent comparative studies reveal a remarkable diversity of song-learning strategies in the songbirds. Here, we discuss recent studies that shed light on the possible functional basis of different song-learning programs. We argue that further insights into the evolution and ecology of song learning will require that comparative data and functional hypotheses be analyzed in a phylogenetic context, and we review recent studies that we feel might be the first steps in this process. PMID- 16701359 TI - Mimulus finds centromeres in the driver's seat. AB - Sandler and Novitski first pointed out in 1957 that chromosomes could selfishly exploit meiotic asymmetries to maximize their own transmission, in a process termed 'meiotic drive'. However, since then, only post-meiotic processes of non Mendelian inheritance have received serious scientific attention in studies of transmission distortion. A recent study by Fishman and Willis puts the focus squarely back on meiotic drive. They found completely biased transmission of a centromere-linked locus from an outcrossing Mimulus species over that from an inbred species, providing the first direct evidence that centromeres can act as general, powerful meiotic drivers. This study suggests that, although difficult to detect experimentally, female meiotic drive is a major evolutionary force in nature. PMID- 16701360 TI - Mind the gap: how open were European primeval forests? AB - There are two competing hypotheses about the structure and dynamics of primeval forests in lowland Europe: the high-forest and the wood-pasture hypotheses, both of which influence current European forest conservation policies. In a recent study using pollen-analytical data from across lowland Europe, Mitchell provides support for rejecting the wood-pasture hypothesis. His study is important for future forest management planning and for showing how hypotheses about large herbivores as biotic factors can be tested using palaeoecological data. PMID- 16701361 TI - Genetic compatibility and sexual selection. PMID- 16701362 TI - Corruption of journal Impact Factors. PMID- 16701363 TI - Conserving taxonomic complexity. AB - Traditional species-based conservation programmes are appropriate in situations where species are readily identifiable. However, in certain taxonomically complex groups of organisms, generally characterized by the presence of uniparental lineages and reticulate evolution, it is not possible to classify biodiversity into discrete and unambiguous species. Attempts to impose species-based conservation on such taxonomically complex groups are proving untenable, and threaten to divert scarce resources and taxonomic expertise from the conservation of other priority groups. We argue here that a new approach should be adopted for taxonomically complex groups. We advocate the conservation of evolutionary processes that generate taxonomic biodiversity, rather than the preservation of a limited number of poorly defined taxa arising from this evolution. PMID- 16701364 TI - The priming of periodical cicada life cycles. AB - Periodical cicadas in the genus Magicicada have unusually long life cycles for insects, with periodicities of either 13 or 17 years. Biologists have explained the evolution of these prime number period lengths in terms of resource limitation, enemy avoidance, hybridization and climate change. Here, I question two aspects of these explanations: that the origin of the life cycles was associated with Pleistocene ice age events, and that they evolved from shorter life cycles through the lengthening of nymphal stages in annual increments. Instead, I suggest that these life cycles evolved earlier than the Pleistocene and involved an abrupt transition from a nine-year to a 13-year life cycle, driven, in part, by interspecific competition. PMID- 16701365 TI - The evolutionary and ecological benefits of asteroid and comet impacts. AB - Commonly viewed solely as agents of destruction, asteroid and comet impact events can also have a beneficial influence on processes from the molecular to the evolutionary scale. On the heavily bombarded early Earth, impacts might have delivered and caused the synthesis of prebiotic compounds that eventually led to life. At the organismal and ecosystem level, impact events can provide new habitats through the shock processing of target materials and by enhancing water availability, such as within intracrater lakes. At the evolutionary level, by destroying entire groups of organisms, impacts might have been instrumental in enabling the rise of new groups, such as the dinosaurs and mammals. Here, we synthesize the emerging literature on the beneficial effects of impacts to provide a novel perspective on these extraterrestrial agents of biological change. PMID- 16701366 TI - The avian genome uncovered. AB - Our knowledge of avian genomics has increased rapidly over the past few years, culminating in the recent publication of a draft sequence of the chicken genome, a milestone event in avian genetics and evolutionary biology. Comparative analysis reveals a compact avian genome structure containing a similar number of genes as found in mammals but with shorter intergenic DNA sequences and fewer repeats. Recombination is at a higher rate than in mammals, particularly for microchromosomes. These also differ from macrochromosomes in their GC and gene content, and their substitution rate. The avian genome has remained unusually stable during evolution and contrasts sharply with the frequent chromosomal rearrangements seen in the rodent lineage. Detailed analyses of polymorphism levels in chickens, including a genome-wide screening in three chicken breeds yielding a set of 2.8 million SNP markers, reveal unexpectedly high levels of genetic diversity. As a notable exception, the female-specific W chromosome is very low in diversity, a probable consequence of the effect of selection on non recombining chromosomes. The chicken genome promises to be a useful resource for ecological and evolutionary studies of other bird species. PMID- 16701367 TI - Information and its use by animals in evolutionary ecology. AB - Information is a crucial currency for animals from both a behavioural and evolutionary perspective. Adaptive behaviour relies upon accurate estimation of relevant ecological parameters; the better informed an individual, the better it can develop and adjust its behaviour to meet the demands of a variable world. Here, we focus on the burgeoning interest in the impact of ecological uncertainty on adaptation, and the means by which it can be reduced by gathering information, from both 'passive' and 'responsive' sources. Our overview demonstrates the value of adopting an explicitly informational approach, and highlights the components that one needs to develop useful approaches to studying information use by animals. We propose a quantitative framework, based on statistical decision theory, for analysing animal information use in evolutionary ecology. Our purpose is to promote an integrative approach to studying information use by animals, which is itself integral to adaptive animal behaviour and organismal biology. PMID- 16701368 TI - Tackling the population genetics of clonal and partially clonal organisms. AB - Many clonal organisms experience occasional events of sexual recombination, with profound consequences for their population dynamics and evolutionary trajectories. With the recent development of polymorphic genetic markers and new statistical methods, we now have an unprecedented ability to detect recombination in organisms that are thought to reproduce strictly, or essentially asexually. However, it is not always obvious which methodology to apply. Consequently, biologists might decide how to analyse their data without clear guidelines. Here, we discuss the available methods, focusing on those best suited when working with limited genetic information, such as a few genetic markers or DNA sequences. We conclude by commenting on the prospects offered by some recent conceptual advances and the access to high throughput technologies in an increasing number of model organisms. PMID- 16701369 TI - Redressing the sex imbalance in knowledge of vector biology. AB - The recent development of transgenic mosquitoes that are resistant to infection by the Plasmodium malarial parasite is a promising new tool in the fight against malaria. However, results of large-scale field releases of alternatively modified mosquitoes carried out during the 1970s and 1980s suggest that this approach could be difficult to implement in the field. These past attempts to control mosquito populations largely floundered as a result of our insufficient understanding of the behavioural ecology of released males. In spite of this, contemporary research on genetic control strategies has concentrated predominantly on molecular aspects, with little progress being made toward resolving key ecological uncertainties, male mosquito ecology being the most important. Here, we review the state of knowledge of male mosquito ecology, and highlight priorities for further research. Case studies of two crop pests, the Mediterranean fruit fly and melon fly, are given as examples of how knowledge of male ecology facilitates successful control in other species. Unless similar information becomes available for mosquitoes, any future genetic control strategy will risk failure. PMID- 16701371 TI - How to manage biological invasions under globalization. AB - Protecting national borders against biological invasions is becoming increasingly difficult because those whose actions result in invasions seldom bear legal responsibility for those actions. Invasion costs are often an externality (an unintended side effect) of international trade. Externalities are best dealt with by internalizing them; that is, by getting those who harm society to meet the cost. This is the 'polluter pays principle', which, under current trade rules, is difficult to implement. Tariffs could, however, be used to confront exporters with the costs of their actions, and the right to do this should be embedded in trade agreements. At the same time, international aid could be used to protect donor societies against the inability of some other countries to take appropriate biosecurity measures. The impact of invasions can thus be reduced by tackling their economic externalities. PMID- 16701372 TI - The politics of assessing risk for biological invasions: the USA as a case study. AB - The current regulation of biological invasions rests on an unwarranted presumption (that the invader will cause no harm) and on risk assessment procedures that are narrowly focused, subjective, often arbitrary and unquantified, and subject to political interference. Although this current approach dominates international treaties and most national policies, it has not stemmed the rising tide of biological invasions, as evidenced by several examples from the USA. Technical advances in measuring and predicting impacts of introduced species will improve risk assessments. Additionally, focusing squarely on the risks associated not only with a proposed species introduction, but also on the goals of the introduction and on alternative ways of achieving them, would lead to more-informed decisions permitting the introduction of a species and fewer problematic invaders. In assessing the alternatives to introductions, the precautionary principle should be given heavy weight, as should the distribution of possible costs and benefits. PMID- 16701373 TI - The role of propagule pressure in explaining species invasions. AB - Human-mediated species invasions are a significant component of current global environmental change. There is every indication that the rate at which locations are accumulating non-native species is accelerating as free trade and globalization advance. Thus, the need to incorporate predictive models in the assessment of invasion risk has become acute. However, finding elements of the invasion process that provide consistent explanatory power has proved elusive. Here, we propose propagule pressure as a key element to understanding why some introduced populations fail to establish whereas others succeed. In the process, we illustrate how the study of propagule pressure can provide an opportunity to tie together disparate research agendas within invasion ecology. PMID- 16701374 TI - Hybridization as an invasion of the genome. AB - Hybridization between species is commonplace in plants, but is often seen as unnatural and unusual in animals. Here, I survey studies of natural interspecific hybridization in plants and a variety of animals. At least 25% of plant species and 10% of animal species, mostly the youngest species, are involved in hybridization and potential introgression with other species. Species in nature are often incompletely isolated for millions of years after their formation. Therefore, much evolution of eventual reproductive isolation can occur while nascent species are in gene-flow contact, in sympatry or parapatry, long after divergence begins. Although the relative importance of geographic isolation and gene flow in the origin of species is still unknown, many key processes involved in speciation, such as 'reinforcement' of post-mating isolation by the evolution of assortative mating, will have ample opportunity to occur in the presence of continuing gene flow. Today, DNA sequence data and other molecular methods are beginning to show that limited invasions of the genome are widespread, with potentially important consequences in evolutionary biology, speciation, biodiversity, and conservation. PMID- 16701375 TI - Emerging pathogens: the epidemiology and evolution of species jumps. AB - Novel pathogens continue to emerge in human, domestic animal, wildlife and plant populations, yet the population dynamics of this kind of biological invasion remain poorly understood. Here, we consider the epidemiological and evolutionary processes underlying the initial introduction and subsequent spread of a pathogen in a new host population, with special reference to pathogens that originate by jumping from one host species to another. We conclude that, although pathogen emergence is inherently unpredictable, emerging pathogens tend to share some common traits, and that directly transmitted RNA viruses might be the pathogens that are most likely to jump between host species. PMID- 16701376 TI - Genes invading new populations: a risk assessment perspective. AB - One of the concerns raised over the introduction of genetically modified crops is that transgenes will invade populations of wild relatives, causing ecologically significant changes in fitness. In recent years, this has given rise to several studies estimating hybridization rates and the fitness of crop-wild relative hybrids. These studies have established that transgenes are likely to move to F1 hybrids, albeit at low frequency. Hybridization, however, is not synonymous with introgression, and questions remain as to whether particular transgenes will cause ecologically significant changes in recipient plant populations. Research effort should now focus on estimating any changes in the fitness of a population as a consequence of having a transgene, understanding genotype x environment interactions, and deducing the extent to which pathogens and herbivores (transgene targets) regulate wild relative populations. This will involve a combination of manipulative experiments and empirically motivated mathematical models. PMID- 16701377 TI - Invasion of the continents: cyanobacterial crusts to tree-inhabiting arthropods. AB - The colonization of continental environments (land and fresh water) has focused historically on a major event during the mid-Paleozoic Era characterized by the relatively sudden emergence of megascopic embryophytes, fungi, arthropods and tetrapods. A significant earlier phase of Precambrian (Archean and Proterozoic Eons) terrestrialization extends to the first 80% of the history of life and records the colonization of subaerial soils or rock surfaces predominantly by cyanobacterial mats and crusts. These two phases are separated by a approximately 90-million-year early Paleozoic interlude of minimal terrestrial colonization. Trophically modern ecosystems appeared during the Late Silurian-Middle Devonian (425-375 million years ago), consisting of complex symbiotic, parasitic and other trophic associations, including detritivory and limited herbivory. The integration of these two historically disparate fields (Precambrian microorganisms and their biochemical and sedimentological signatures, and the paleoecology of mid-Paleozoic ecosystems) has resulted in a wider perspective on terrestrialization. Here, I present an ecological and evolutionary context for the emergence of terrestrial ecosystems and examine associations among organisms, from the endosymbiotic capture of organelles by eukaryotes to modes of metazoan nutrition on land. Such studies now enable the tracking, in ecological detail, of the invasion of continental environments during the past 3.5 billion years of life. PMID- 16701378 TI - The invasion of language: emergence, change and death. AB - Research into the emergence and evolution of human language has received unprecedented attention during the past 15 years. Efforts to better understand the processes of language emergence and evolution have proceeded in two main directions: from the top-down (linguists) and from the bottom-up (cognitive scientists). Language can be viewed as an invading process that has had profound impact on the human phenotype at all levels, from the structure of the brain to modes of cultural interaction. In our view, the most effective way to form a connection between the two efforts (essential if theories for language evolution are to reflect the constraints imposed on language by the brain) lies in computational modelling, an approach that enables numerous hypotheses to be explored and tested against objective criteria and which suggest productive paths for empirical researchers to then follow. Here, with the aim of promoting the cross-fertilization of ideas across disciplines, we review some of the recent research that has made use of computational methods in three principal areas of research into language evolution: language emergence, language change, and language death. PMID- 16701379 TI - Testing evolutionary theories of aging in wild populations. AB - Classic theories for the evolution of senescence predict that rates of aging should be highest in populations in which extrinsic mortality is high. This predication is called into question in new work by David Reznick and co-workers, who found that guppies Poecilia reticulata derived from natural populations with high levels of predation live the longest in the laboratory. This study illustrates that the effect of mortality on aging might depend on how we define aging, and on the particular cause of increased mortality. PMID- 16701380 TI - Do bushmeat consumers have other fish to fry? AB - The overexploitation of tropical forests for bushmeat and of the oceans for fish are two of the most important threats to global biodiversity. Both phenomena also have manifold implications for human livelihoods and food security. A recent article by Brashares et al. indicates that these two resources are linked: when regional fish supplies are low, local bushmeat hunting intensifies. Although coordinated management of bushmeat and fisheries is thus needed, gaps in our knowledge of these systems must first be filled. PMID- 16701381 TI - Intimidating butterflies. AB - Many butterfly species feature two or four conspicuously coloured eyespots on their wings; the most commonly heard explanation for these markings is that they are used to intimidate predators. However, this explanation has stood, until recently, on the flimsiest of empirical foundations. Now, Adrian Vallin and colleagues have demonstrated empirically the effectiveness of these signals in dissuading avian predators from attacking the bearers. Their work suggests that predators can be intimidated by bold displaying by otherwise defenceless prey, and opens up an interesting challenge to evolutionary ecologists to explain how this apparently maladaptive behaviour is retained in predator populations. PMID- 16701382 TI - The reality of taxonomic change. PMID- 16701384 TI - Beyond Singapore: Hong Kong and Asian biodiversity. PMID- 16701386 TI - Genomics-fueled approaches to current challenges in marine ecology. AB - As a result of recent advances in genomic technology, a 'tool set' is emerging that can be used by marine ecologists to gain new perspectives on central questions in the field. Techniques such as gene expression profiling using DNA microarrays, when placed in an ecological context, stand to advance greatly our understanding of how organisms respond to abiotic and biotic stresses. Here, we target areas in which adding genomics to ecological and physiological investigations will significantly advance our understanding of crucial issues ranging from the general biological effects of environmental temperature changes on individuals and communities to the interactions between symbionts in coral bleaching. At a time when both short- and long-term perturbations of marine ecosystems are increasing in severity, this kind of integrative approach could generate new and exciting hypotheses about the functioning of marine organisms and communities. PMID- 16701387 TI - Genome duplication, extinction and vertebrate evolution. AB - Vertebrate evolution has been punctuated by three episodes of widespread gene or genome duplication, which have been linked with the origin of vertebrates, gnathostomes and teleosts, respectively. These three events coincide with bursts of character acquisition and increases in phenotypic complexity, and many researchers have suggested a causal relationship between the two. However, this pattern is derived from data for living taxa only; we argue here that, when fossils are taken into account, bursts of character acquisition disappear and gen(om)e duplication in vertebrate phylogeny can no longer be correlated with the origin of body plans. If patterns of character acquisition or morphological gaps between higher taxa are a reflection of phenotypic complexity, then more inclusive data sets incorporating fossil taxa provide no support for hypotheses linking gen(om)e duplications and the evolution of complexity in vertebrates. PMID- 16701388 TI - Molecular dating when rates vary. AB - Molecular-dating techniques potentially enable us to estimate the time of origin of any biological lineage. Such techniques were originally premised on the assumption of a 'molecular clock'; that is, the assumption that genetic change accumulated steadily over time. However, it is becoming increasingly clear that constant rates of molecular evolution might be the exception rather than the rule. Recently, new methods have appeared that enable the incorporation of variable rates into molecular dating. Direct comparisons between these methods are difficult, because they differ in so many respects. However, the assumptions about rate change on which they rely fall into a few broad categories. Improving our understanding of molecular evolution will be an important next step towards evaluating and improving these methods. PMID- 16701389 TI - Spatial epidemiology: an emerging (or re-emerging) discipline. AB - Spatial epidemiology is the study of spatial variation in disease risk or incidence. Several ecological processes can result in strong spatial patterns of such risk or incidence: for example, pathogen dispersal might be highly localized, vectors or reservoirs for pathogens might be spatially restricted, or susceptible hosts might be clumped. Here, we briefly describe approaches to spatial epidemiology that are spatially implicit, such as metapopulation models of disease transmission, and then focus on research in spatial epidemiology that is spatially explicit, such as the creation of risk maps for particular geographical areas. Although the spatial dynamics of infectious diseases are the subject of intensive study, the impacts of landscape structure on epidemiological processes have so far been neglected. The few studies that demonstrate how landscape composition (types of elements) and configuration (spatial positions of those elements) influence disease risk or incidence suggest that a true integration of landscape ecology with epidemiology will be fruitful. PMID- 16701390 TI - Climate change and marine plankton. AB - Understanding how climate change will affect the planet is a key issue worldwide. Questions concerning the pace and impacts of climate change are thus central to many ecological and biogeochemical studies, and addressing the consequences of climate change is now high on the list of priorities for funding agencies. Here, we review the interactions between climate change and plankton communities, focusing on systematic changes in plankton community structure, abundance, distribution and phenology over recent decades. We examine the potential socioeconomic impacts of these plankton changes, such as the effects of bottom-up forcing on commercially exploited fish stocks (i.e. plankton as food for fish). We also consider the crucial roles that plankton might have in dictating the future pace of climate change via feedback mechanisms responding to elevated atmospheric CO(2) levels. An important message emerges from this review: ongoing plankton monitoring programmes worldwide will act as sentinels to identify future changes in marine ecosystems. PMID- 16701391 TI - Network thinking in ecology and evolution. AB - Although pairwise interactions have always had a key role in ecology and evolutionary biology, the recent increase in the amount and availability of biological data has placed a new focus on the complex networks embedded in biological systems. The increased availability of computational tools to store and retrieve biological data has facilitated wide access to these data, not just by biologists but also by specialists from the social sciences, computer science, physics and mathematics. This fusion of interests has led to a burst of research on the properties and consequences of network structure in biological systems. Although traditional measures of network structure and function have started us off on the right foot, an important next step is to create biologically realistic models of network formation, evolution, and function. Here, we review recent applications of network thinking to the evolution of networks at the gene and protein level and to the dynamics and stability of communities. These studies have provided new insights into the organization and function of biological systems by applying existing techniques of network analysis. The current challenge is to recognize the commonalities in evolutionary and ecological applications of network thinking to create a predictive science of biological networks. PMID- 16701392 TI - Vertebrate ecology and evolution in 'Deep Time'. PMID- 16701393 TI - The importance in fishery management of leaving the big ones. AB - Research by Berkeley et al. and by Bobko and Berkeley has recently demonstrated that older individuals of some fish species produce larvae that have substantially better survival potential than do larvae from younger fishes. These new findings augment established knowledge that larger individuals usually have exponentially greater fecundity. This is important because commercial fisheries and especially recreational fishing often target the larger fish. The protection of larger or older individuals is necessary for the sustainability of species currently exploited by humans. PMID- 16701394 TI - Hybridization, transgressive segregation, genetic covariation, and adaptive radiation. AB - Analysis of adaptive radiation has had a central role in the development of evolutionary theory, but it is not clear why some groups radiate and others do not. Two recent papers by Albertson and colleagues on the genetic architecture of East African cichlid fishes implicate hybridization, transgressive segregation and genetic covariation in the early stages of adaptive radiation. Transgressive segregation and genetic covariation might be key innovations in genetic architecture that favor adaptive radiation. PMID- 16701396 TI - Data sharing in ecology and evolution. PMID- 16701395 TI - Organisms in nature as a central focus for biology. PMID- 16701397 TI - A new eusocial vertebrate? PMID- 16701398 TI - Buzziness as usual? Questioning the global pollination crisis. AB - Concerns have been raised that invertebrate pollinators of crops and wild plants are in decline as a result of modern agricultural practices, habitat degradation, and introduced pests and diseases. This has led to demands for a response by land managers, conservationists and political decision makers to the impending 'global pollinator crisis'. In questioning this crisis, it becomes apparent that perceptions of a pollinator crisis are driven mainly by reported declines of crop pollinating honeybees in North America, and bumblebees and butterflies in Europe, whereas native pollinator communities elsewhere show mixed responses to environmental change. Additionally, few staple food crops depend on pollinator services, and most crops that do are grown at small scales in diversified agro ecosystems that are likely to support healthy pollinator communities, or in highly managed systems that are largely independent of wild pollinators. Consequently, justifying conservation action on the basis of deteriorating pollinator services might be misplaced. Nevertheless, existing initiatives to monitor pollinators are well founded, given the uncertainty about the dynamics of pollinator populations. PMID- 16701399 TI - 'Haldane's Sieve' in a metapopulation: sifting through plant reproductive polymorphisms. AB - An important result of population genetics is that advantageous mutations will be fixed by selection in a population with a greater probability if they are dominant rather than recessive. This selective filter on new variants entering a population, termed 'Haldane's Sieve', has hitherto been invoked to account for the greater role of dominant than completely recessive mutations in adaptive evolution. Here, we suggest that a process similar to Haldane's Sieve will act on migrants into subpopulations of a metapopulation, and that the repeated action of Haldane's Sieve on alleles maintained by frequency-dependent selection, such as those responsible for many plant reproductive polymorphisms, is expected to bias their frequency distribution in favour of dominant alleles. The genetic and phenotypic signatures left by these processes might provide additional indirect support for the contentious idea that metapopulation dynamics have had an important role in shaping the ecology and evolution of some plant species. PMID- 16701400 TI - New paradigms for supporting the resilience of marine ecosystems. AB - Resource managers and scientists from disparate disciplines are rising to the challenge of understanding and moderating human impacts on marine ecosystems. Traditional barriers to communication between marine ecologists, fisheries biologists, social scientists and economists are beginning to break down, and the distinction between applied and basic research is fading. These ongoing trends arise, in part, from an increasing awareness of the profound influence of people on the functioning of all marine ecosystems, an increased focus on spatial and temporal scale, and a renewed assessment of the role of biodiversity in the sustainability of ecosystem goods and services upon which human societies depend. Here, we highlight the emergence of a complex systems approach for sustaining and repairing marine ecosystems, linking ecological resilience to governance structures, economics and society. PMID- 16701401 TI - Fire as a global 'herbivore': the ecology and evolution of flammable ecosystems. AB - It is difficult to find references to fire in general textbooks on ecology, conservation biology or biogeography, in spite of the fact that large parts of the world burn on a regular basis, and that there is a considerable literature on the ecology of fire and its use for managing ecosystems. Fire has been burning ecosystems for hundreds of millions of years, helping to shape global biome distribution and to maintain the structure and function of fire-prone communities. Fire is also a significant evolutionary force, and is one of the first tools that humans used to re-shape their world. Here, we review the recent literature, drawing parallels between fire and herbivores as alternative consumers of vegetation. We point to the common questions, and some surprisingly different answers, that emerge from viewing fire as a globally significant consumer that is analogous to herbivory. PMID- 16701402 TI - Fifty millennia of catastrophic extinctions after human contact. AB - Debate continues to rage between enthusiasts for climate change versus humans as a cause of the catastrophic faunal extinctions that have occurred in the wake of human arrival in previously uninhabited regions of the world. A global pattern of human arrival to such landmasses, followed by faunal collapse and other ecological changes, appears without known exception. This strongly suggests to some investigators that a more interesting extinction debate lies within the realm of potential human-caused explanations and how climate might exacerbate human impacts. New observations emerging from refined dating techniques, paleoecology and modeling suggest that the megafaunal collapses of the Americas and Australia, as well as most prehistoric island biotic losses, trace to a variety of human impacts, including rapid overharvesting, biological invasions, habitat transformation and disease. PMID- 16701403 TI - Body size in ecological networks. AB - Body size determines a host of species traits that can affect the structure and dynamics of food webs, and other ecological networks, across multiple scales of organization. Measuring body size provides a relatively simple means of encapsulating and condensing a large amount of the biological information embedded within an ecological network. Recently, important advances have been made by incorporating body size into theoretical models that explore food web stability, the patterning of energy fluxes, and responses to perturbations. Because metabolic constraints underpin body-size scaling relationships, metabolic theory offers a potentially useful new framework within which to develop novel models to describe the structure and functioning of ecological networks and to assess the probable consequences of biodiversity change. PMID- 16701404 TI - Ernst Mayr, a retrospective. PMID- 16701405 TI - Hiding in plain sight. AB - Although the principles of disruptive colouration are widely believed to explain a variety of animal colour patterns, there has been no field evidence that it works to reduce the detection rates of natural prey. In a recent paper, Cuthill et al. successfully address this shortfall, separating the benefits of background matching from those of disruptive colouration. Their results provide the first definitive field support for this long-recognized phenomenon and suggest several new avenues of research. PMID- 16701406 TI - Are scared prey as good as dead? AB - Predators affect prey and their resources by changing the density and traits (e.g. morphology and behavior) of those prey. Ecological studies and models of community dynamics, however, typically only incorporate how changes in prey densities, rather than their traits, affect community dynamics. In a recent meta analysis, Preisser et al. show that trait effects are as large, if not larger than density effects. This strongly suggests that trait effects should be integrated into empirical and theoretical studies. PMID- 16701407 TI - The future fate of the Antarctic marine biota? PMID- 16701408 TI - Female songbirds still struggling to be heard. PMID- 16701409 TI - Emerging pathogens: fungal host jumps following anthropogenic introduction. PMID- 16701410 TI - Sense and stability in animal names. PMID- 16701411 TI - Evidence-based conservation: dealing with social issues. PMID- 16701413 TI - Biodiversity conservation and the extinction of experience. AB - Biodiversity loss is a matter of great concern among conservation scientists, but the wherewithal to reverse this trend is generally lacking. One reason is that nearly half of the world's people live in urban areas and are increasingly disconnected from nature. If there is to be broad-based public support for biodiversity conservation, the places where people live and work should be designed so as to provide opportunities for meaningful interactions with the natural world. Doing so has the potential not only to engender support for protecting native species, but also to enhance human well-being. Accomplishing these goals will necessitate conservation scientists forging new collaborations with design professionals, health practitioners and social scientists, as well as encouraging the participation of the general public. PMID- 16701414 TI - Adaptation and speciation: what can F(st) tell us? AB - A useful way of summarizing genetic variability among different populations is through estimates of the inbreeding coefficient, F(st). Several recent studies have tried to use the distribution of estimates of F(st) from individual genetic loci to detect the effects of natural selection. However, the promise of this approach has yet to be fully realized owing to the pervasive dogma that this distribution is highly dependent on demographic history. Here, I review recent theoretical results that indicate that the distribution of estimates of F(st) is generally expected to be robust to the vagaries of demographic history. I suggest that analyses based on it provide a useful first step for identifying candidate genes that might be under selection, and explore the ways in which this information can be used in ecological and evolutionary studies. PMID- 16701415 TI - Facing herbivory as you grow up: the ontogeny of resistance in plants. AB - As plants develop from seeds to seedlings, juveniles and mature stages, their ontogeny can constrain the expression of resistance to herbivore damage. Nevertheless, ecological and evolutionary theories regarding interactions between plants, herbivores and their natural enemies are largely based on observations and experiments conducted at a single ontogenetic stage. Owing to resource allocation and architectural constraints in plants, and the influence of herbivore foraging behavior, resistance to herbivores is likely to change during plant development. We propose that such changes are likely to occur in a non linear fashion and suggest that the role of ontogeny should be incorporated as an important factor in new syntheses of plant defense theory. PMID- 16701416 TI - Consensus decision making in animals. AB - Individual animals routinely face decisions that are crucial to their fitness. In social species, however, many of these decisions need to be made jointly with other group members because the group will split apart unless a consensus is reached. Here, we review empirical and theoretical studies of consensus decision making, and place them in a coherent framework. In particular, we classify consensus decisions according to the degree to which they involve conflict of interest between group members, and whether they involve either local or global communication; we ask, for different categories of consensus decision, who makes the decision, what are the underlying mechanisms, and what are the functional consequences. We conclude that consensus decision making is common in non-human animals, and that cooperation between group members in the decision-making process is likely to be the norm, even when the decision involves significant conflict of interest. PMID- 16701417 TI - Biogeography and evolution of the genus Homo. AB - The debate about the origins of modern humans has traditionally focussed on two contrasting views. Multi-regional evolution proposes that present-day populations worldwide are the descendants of in situ evolution after an initial dispersal of Homo erectus from Africa during the Lower Pleistocene. The alternative, Out-of Africa 2, proposes that all present-day populations are descended from a recent common ancestor that lived in East Africa approximately 150 000 years ago, the population of which replaced all regional populations. The weight of the evidence is now in favour of Out-of-Africa 2, and discussion is now dominated by the causes of the dispersal of modern humans out of Africa and the outcome of contact with other populations. Fresh approaches, from disciplines hitherto peripheral to the debate, such as evolutionary ecology, and new discoveries are challenging established views, particularly the prevalent idea that biologically superior modern humans were the cause of the demise of all other populations of Homo worldwide. Climate-driven ecological change has been, as with many other taxa, the driving force in the geographical range dynamics of the genus Homo. PMID- 16701418 TI - New insights into the evolutionary history of photoreceptor cells. AB - Although the common descent of all life has been widely accepted since Darwin's time, new research occasionally provides us with arresting reminders of the unity of evolutionary history. Recent papers by Arendt et al. and Panda et al. provide one such reminder. They illustrate that the two classes of animal photoreceptors, ciliary and rhabdomeric photoreceptors, are likely to share an ancient common ancestor and have been evolving in parallel since their duplication over 600 million years ago. PMID- 16701419 TI - Genetic variability under mutation selection balance. AB - A fundamental problem in evolutionary genetics is understanding how high levels of genetic variation in quantitative traits are maintained in natural populations. Variation is removed by the natural selection of individuals with optimal phenotypes and is recovered by mutation; however, previous analyses had indicated that a mutation-selection balance was insufficient to maintain observed levels of genetic variation in these traits. Using more general models, however, it has recently been shown that it is indeed a sufficient mechanism. These models can be used to explore other phenomena in evolutionary biology. PMID- 16701420 TI - Are invasive species the drivers of ecological change? AB - Invasive species are widely accepted as one of the leading direct causes of biodiversity loss. However, much of the evidence for this contention is based on simple correlations between exotic dominance and native species decline in degraded systems. Although appealing, direct causality is not the only possible interpretation. A plausible alternative hypothesis is that exotic dominance could be the indirect consequence of habitat modification driving native species loss. In a new paper, MacDougall and Turkington now provide the first direct test of whether invasive species are the drivers of community change, or merely 'passengers' along for the environmental ride. PMID- 16701421 TI - Is science as global as we think? PMID- 16701422 TI - It's a puzzle all right: the hippocampus and food hoarding. PMID- 16701424 TI - Evolution of phenotypic plasticity: where are we going now? AB - The study of phenotypic plasticity has progressed significantly over the past few decades. We have moved from variation for plasticity being considered as a nuisance in evolutionary studies to it being the primary target of investigations that use an array of methods, including quantitative and molecular genetics, as well as of several approaches that model the evolution of plastic responses. Here, I consider some of the major aspects of research on phenotypic plasticity, assessing where progress has been made and where additional effort is required. I suggest that some areas of research, such the study of the quantitative genetic underpinning of plasticity, have been either settled in broad outline or superseded by new approaches and questions. Other issues, such as the costs of plasticity are currently at the forefront of research in this field, and are likely to be areas of major future development. PMID- 16701425 TI - Orchid diversity: an evolutionary consequence of deception? AB - The Orchidaceae are one of the most species-rich plant families and their floral diversity and pollination biology have long intrigued evolutionary biologists. About one-third of the estimated 18,500 species are thought to be pollinated by deceit. To date, the focus has been on how such pollination evolved, how the different types of deception work, and how it is maintained, but little progress has been made in understanding its evolutionary consequences. To address this issue, we discuss here how deception affects orchid mating systems, the evolution of reproductive isolation, speciation processes and neutral genetic divergence among species. We argue that pollination by deceit is one of the keys to orchid floral and species diversity. A better understanding of its evolutionary consequences could help evolutionary biologists to unravel the reasons for the evolutionary success of orchids. PMID- 16701426 TI - Hybridization, glaciation and geographical parthenogenesis. AB - Parthenogenetic organisms are all female and reproduce clonally. The transition from sex to parthenogenesis is frequently associated with a major change in geographical distribution, often biasing parthenogenetic lineages towards environments that were severely affected by the glacial cycles of the Late Pleistocene. It is difficult to interpret these patterns as arising simply as a result of selection for the demographic effects of parthenogenesis because many parthenogenetic organisms are also hybrids. Here, I argue that many cases of geographical parthenogenesis might be best seen as part of a broader pattern of hybrid advantage in new and open environments. Parthenogenesis in these cases could have a more secondary role of stabilizing strongly selected hybrid genotypes. In this context, geographical parthenogenesis might tell us more about the role of hybridization in evolution than about the role of sex. PMID- 16701427 TI - Using the satellite-derived NDVI to assess ecological responses to environmental change. AB - Assessing how environmental changes affect the distribution and dynamics of vegetation and animal populations is becoming increasingly important for terrestrial ecologists to enable better predictions of the effects of global warming, biodiversity reduction or habitat degradation. The ability to predict ecological responses has often been hampered by our rather limited understanding of trophic interactions. Indeed, it has proven difficult to discern direct and indirect effects of environmental change on animal populations owing to limited information about vegetation at large temporal and spatial scales. The rapidly increasing use of the Normalized Difference Vegetation Index (NDVI) in ecological studies has recently changed this situation. Here, we review the use of the NDVI in recent ecological studies and outline its possible key role in future research of environmental change in an ecosystem context. PMID- 16701428 TI - Should we expect population thresholds for wildlife disease? AB - Host population thresholds for the invasion or persistence of infectious disease are core concepts of disease ecology and underlie disease control policies based on culling and vaccination. However, empirical evidence for these thresholds in wildlife populations has been sparse, although recent studies have begun to address this gap. Here, we review the theoretical bases and empirical evidence for disease thresholds in wildlife. We see that, by their nature, these thresholds are rarely abrupt and always difficult to measure, and important facets of wildlife ecology are neglected by current theories. Empirical studies seeking to identify disease thresholds in wildlife encounter recurring obstacles of small sample sizes and confounding factors. Disease control policies based solely on threshold targets are rarely warranted, but management to reduce abundance of susceptible hosts can be effective. PMID- 16701429 TI - Deception and the origin of honest signals. AB - Deceptive signals are a challenge to explain because on average, signals should be reliable. When being deceived is costly to the receiver, a coevolutionary struggle between senders and receivers can ensue. Recent work by Macias Garcia and Ramirez raises the intriguing possibility that through such a coevolutionary process, cheats can become honest. PMID- 16701430 TI - Predictive adaptive responses and human evolution. AB - The importance of a single genotype being able to produce different phenotypes in different environments (phenotypic plasticity) is widely recognized in evolutionary theory and its adaptive significance is clear. In most cases, the developing organism responds to an environmental cue by producing a selectively and immediately appropriate phenotype. One subset of phenotypic responses to environmental stimuli, however, does not necessarily provide an immediate selective advantage. Rather, these kinds of responses, which we call 'predictive adaptive responses' (PARs), act primarily to improve fitness at a later stage of development. We argue that PARs have had an important role in human evolution, and that their recognition and interpretation has major significance for public health. PMID- 16701431 TI - On the origin of Antarctic marine benthic community structure. AB - Environmental conditions fostering marine communities around Antarctica differ fundamentally from those in the rest of the world's oceans, particularly in terms of pronounced climatic fluctuations and extreme cold. Here, we argue that the rarity of pelagic larval stages in Antarctic marine benthic invertebrate species is a consequence of evolutionary temperature adaptation and that this has greatly contributed to the current structure of the Antarctic benthic community. In arguing this position, we challenge the likelihood of previously suggested survival strategies of benthic communities on the Antarctic continental shelf and slope during Cenozoic glacial periods. By integrating evidence from marine geology and geophysics, we suggest that the Antarctic continental shelf and slope were both unfavourable environments for benthic communities during glacial periods and that community survival was only possible in the deep sea or in shelters on the continental shelf as a result of the diachronism in maximum ice extent. PMID- 16701432 TI - Assessing ancient DNA studies. AB - The study of ancient DNA has the potential to make significant and unique contributions to ecology and evolution. However, the techniques used contain inherent problems, particularly with regards to the generation of authentic and useful data. The solution currently advocated to reduce contamination and artefactual results is to adopt criteria for authentication. Nevertheless, these criteria are not foolproof, and we believe that they have, in practice, replaced the use of thought and prudence when designing and executing ancient DNA studies. We argue here that researchers in this field must take a more cognitive and self critical approach. Specifically, in place of checking criteria off lists, researchers must explain, in sufficient enough detail to dispel doubt, how the data were obtained, and why they should be believed to be authentic. PMID- 16701433 TI - Evolutionary biology of cancer. AB - Cancer is driven by the somatic evolution of cell lineages that have escaped controls on replication and by the population-level evolution of genes that influence cancer risk. We describe here how recent evolutionary ecological studies have elucidated the roles of predation by the immune system and competition among normal and cancerous cells in the somatic evolution of cancer. Recent analyses of the evolution of cancer at the population level show how rapid changes in human environments have augmented cancer risk, how strong selection has frequently led to increased cancer risk as a byproduct, and how anticancer selection has led to tumor-suppression systems, tissue designs that slow somatic evolution, constraints on morphological evolution and even senescence itself. We discuss how applications of the tools of ecology and evolutionary biology are poised to revolutionize our understanding and treatment of this disease. PMID- 16701434 TI - Tropical forests in a changing environment. AB - Understanding and mitigating the impact of an ever-increasing population and global economic activity on tropical forests is one of the great challenges currently facing biologists, conservationists and policy makers. Tropical forests currently face obvious regional changes, both negative and positive, and uncertain global changes. Although deforestation rates have increased to unprecedented levels, natural secondary succession has reclaimed approximately 15% of the area deforested during the 1990s. Governments have also protected 18% of the remaining tropical moist forest; however, unsustainable hunting continues to threaten many keystone mammal and bird species. The structure and dynamics of old-growth forests appear to be rapidly changing, suggesting that there is a pantropical response to global anthropogenic forcing, although the evidence comes almost exclusively from censuses of tree plots and is controversial. Here, I address ongoing anthropogenic change in tropical forests and suggest how these forests might respond to increasing anthropogenic pressure. PMID- 16701435 TI - Genes, brains and mammalian social bonds. AB - Recent studies of monogamous species have revealed the role of the neuropeptides oxytocin and vasopressin in activating reward mechanisms of the brain that are involved in establishing partner recognition and selective 'bonding'. The evolutionary history of these findings resides, at a mechanistic level, in the reciprocal bonding between mother and infant that is common to all mammals. However, in Old World primates, where mother and infant alone would not survive, living in large social groups brings extended family relationships and provides for alloparenting. This has required the emancipation of parenting behaviour from the constraints of hormonal state and the evolution of large brains for decision making that was previously restricted and determined by hormonal state. How this has been achieved, what conserved mechanisms underpin social bonding, and what genetic and mechanistic changes have occurred in the evolution of social bonds are the issues addressed here. PMID- 16701436 TI - Using community viability analysis to identify fragile systems and keystone species. AB - Owing to interdependences among species in ecological communities, the loss of one species can trigger a cascade of secondary extinctions with potentially dramatic effects on the functioning and stability of the community. It is, therefore, important to assess the risk and likely extent of secondary extinctions. Community viability analysis is a new technique that can be used to accomplish this goal. The analysis can also be used to identify fragile community structures and keystone species and, hence, to provide guidelines for conservation priorities. Here, we describe the principles underlying community viability analysis and review its contributions to our understanding of the response of ecological communities to species loss. PMID- 16701437 TI - Something old, something transgenic, or something fungal for mosquito control? AB - In spite of the current research emphasis on the use of transgenic mosquitoes, insecticides are still the main method for controlling malarial mosquitoes. Although pyrethroids are the compounds of choice, insecticide resistance is now threatening the effective life of these invaluable compounds. Two recent studies have re-focused interest on entomopathogenic fungi as useful alternatives to conventional insecticides, suggesting that these fungi could be used as alternative control methods, which would thus also prolong the effective lifetime of pyrethroids. PMID- 16701438 TI - Cascading effects of over-fishing marine systems. AB - Profound indirect ecosystem effects of over-fishing have been shown for coastal systems such as coral reefs and kelp forests. A new study from the ecosystem off the Canadian east coast now reveals that the elimination of large predatory fish can also cause marked cascading effects on the pelagic food web. Overall, the view emerges that, in a range of marine ecosystems, the effects of fisheries extend well beyond the collapse of fish exploited stocks. PMID- 16701439 TI - The darting game in snails and slugs. AB - Love darts are hard 'needles' that many snails and slugs use to pierce their partner during mating. In a few species, darts have been shown to play a role in sperm competition. Two new papers, by Davison et al., and Koene and Schulenburg, might further pique researchers' interest, because they show how the full potential of darts can be tapped for studies of sexual selection in hermaphrodites. PMID- 16701440 TI - Linking nectar amino acids to fitness in female butterflies. AB - Nectar of butterfly-pollinated flowers contains generally higher levels of amino acids than does nectar of flowers pollinated by most other animal types. One proposed explanation is that these amino acids promote butterfly fitness, although the evidence has been equivocal. In a new study, Mevi-Schutz and Erhardt showed that nectar amino acids enhanced fecundity in the butterfly Araschnia levana, but only when the larval diet was poor. Their results support the hypothesis that butterflies are agents of selection for higher nectar amino acid production, suggest that the larval food plant has a key role in the evolution of the flower-butterfly mutualism, and demonstrate that the importance, to butterfly reproduction, of different nutrient sources varies with butterfly nutritional state. PMID- 16701441 TI - Genome duplication and the origin of angiosperms. AB - Despite intensive research, little is known about the origin of the angiosperms and their rise to ecological dominance during the Early Cretaceous. Based on whole-genome analyses of Arabidopsis thaliana, there is compelling evidence that angiosperms underwent two whole-genome duplication events early during their evolutionary history. Recent studies have shown that these events were crucial for the creation of many important developmental and regulatory genes found in extant angiosperm genomes. Here, we argue that these ancient polyploidy events might have also had an important role in the origin and diversification of the angiosperms. PMID- 16701442 TI - The complex business of survival by aposematism. AB - The theory of warning signals dates back to Wallace but is still confusing, controversial and complex. Because predator avoidance of warningly coloured prey (aposematism) is based upon learning and reinforcement, it is difficult to understand how initially rare conspicuous forms subsequently become common. Here, we discuss several possible resolutions to this apparent paradox. Many of these ideas have been largely ignored as a result of implicit assumptions about predator behaviour and assumed lack of variation in the predators, prey and the predation process. Considering the spatial and temporal variation in and mechanisms of behaviour of both predators and prey will make it easier to understand the process and evolution of aposematism. PMID- 16701443 TI - The second wave of evolutionary economics in biology. AB - Several core issues in economics and biology overlap substantially. At the theoretical level, these include analogies and differences among rational choice, learning, genetic evolution and cultural evolution. At the empirical level, they include the structure of decision making, its neural basis and, more generally, human nature. We illustrate here the increasingly important collaboration between economics and biology with several characteristic examples, including signaling, markets, statistical reasoning, cooperation, punishment, reputation and social norms. In contrast to the mutual borrowing of ideas during the 1970s and 1980s, we now see the joint exploration of empirical and theoretical issues by biologists and economists that constitutes a second wave of interactions between the two disciplines. PMID- 16701444 TI - Bridging the gaps between agricultural policy, land-use and biodiversity. AB - The fate of biodiversity is intimately linked to agricultural development. Policy reform is an important driver of changes in agricultural land-use, but there is considerable spatial variation in response to policy and its potential impact on biodiversity. We review the links between policy, land-use and biodiversity and advocate a more integrated approach. Ecologists need to recognize that wildlife friendly farming is not the only land-use strategy that can be used to conserve biodiversity and to research alternative options such as land sparing. There is also a need for social scientists and ecologists to bring their approaches together, so that land-use change and its consequences can be investigated in a more holistic way. PMID- 16701445 TI - Linking aboveground and belowground interactions via induced plant defenses. AB - Plants have a variety of chemical defenses that often increase in concentration following attack by herbivores. Such induced plant responses can occur aboveground, in the leaves, and also belowground in the roots. We show here that belowground organisms can also induce defense responses aboveground and vice versa. Indirect defenses are particularly sensitive to interference by induced feeding activities in the other compartment, and this can disrupt multitrophic interactions. Unravelling the involvement of induced plant responses in the interactions between aboveground and belowground communities associated with plants is likely to benefit from comprehensive metabolomic analyses. Such analyses are likely to contribute to a better understanding of the costs and benefits involved in the selection for induced responses in plants. PMID- 16701446 TI - Linking aboveground and belowground diversity. AB - Aboveground and belowground species interactions drive ecosystem properties at the local scale, but it is unclear how these relationships scale-up to regional and global scales. Here, we discuss our current knowledge of aboveground and belowground diversity links from a global to a local scale. Global diversity peaks towards the Equator for large, aboveground organisms, but not for small (mainly belowground) organisms, suggesting that there are size-related biodiversity gradients in global aboveground-belowground linkages. The generalization of aboveground-belowground diversity relationships, and their role in ecosystem functioning, requires surveys at scales that are relevant to the organisms and ecosystem properties. Habitat sizes and diversity gradients can differ significantly between aboveground and belowground organisms and between ecosystems. These gradients in biodiversity and plant community trait perception need to be acknowledged when studying aboveground-belowground biodiversity linkages. PMID- 16701447 TI - A temporal approach to linking aboveground and belowground ecology. AB - Ecologists are becoming increasingly aware of the role of aboveground-belowground relationships in controlling ecosystem processes and properties. Here, we review recent studies that show that relationships between aboveground and belowground communities operate over a hierarchy of temporal scales, ranging from days to seasons, to millennia, with differing consequences for ecosystem structure and function. We propose that a temporal framework is crucial to our understanding of the nature and ecological significance of relationships between aboveground and belowground communities. PMID- 16701448 TI - Can a genome change its (hot)spots? AB - A new study by Jeffreys et al. shows that the rate of recombination in recombination hotspots in humans is not constant through time. This observation adds weight to the idea that hotspots are transient on evolutionary timescales. However, questions remain as to what controls their evolution and how these rapid changes influence broad-scale rates of recombination. PMID- 16701449 TI - When bigger is better: the need for Amazonian mega-reserves. AB - The rate of forest destruction has accelerated sharply in Brazilian Amazonia, but there are also vital conservation opportunities with the ongoing designation of important new protected areas. In a timely paper, Carlos Peres argues that an extensive network of mega-reserves, operationally defined as those exceeding 1 million ha in area, is needed to ensure the long-term persistence of Amazonian species and ecological processes. Although such protected areas might seem excessively large to some, disparate lines of evidence suggest that mega-reserves are vital for the future of Amazonian biodiversity. PMID- 16701450 TI - Consensus on climate change. PMID- 16701451 TI - Eusociality, menopause and information in matrilineal whales. PMID- 16701452 TI - Pollinator diversity and crop pollination services are at risk. PMID- 16701454 TI - Comparative evaluation and its implications for mate choice. AB - Experiments on decision making by humans show that the choices that we make can be very labile. The magnitude of our preferences, and even our rank ordering of options, can vary according to the number and type of alternatives available for comparison. This apparent irrationality has been argued to result from our use of decision heuristics that have evolved to enable us to choose quickly and efficiently between options differing in multiple attributes. Here, we argue that, because there is also selective pressure for animals to make mating decisions quickly, and because potential mates also differ in multiple attributes, similar decision heuristics might have evolved for mate choice. Following this reasoning, the attractiveness of a given mate will depend on the others with whom he or she is being compared, rather than being an absolute function of his or her underlying quality. We describe some of the ramifications of such comparative evaluation, and argue that it could offer new insights into some of the biggest outstanding problems in mate choice and sexual selection. PMID- 16701455 TI - Optimal immune responses: immunocompetence revisited. AB - The function of the immune system of an animal is to provide defence against infection, in order to maximize fitness. Understanding this and, particularly, how limiting resources are traded off between costly immune responses and other physiological demands, is central to properly understanding life-history traits and their evolution. Here, we propose that functional (rather than immunological) measures of immune responses should be used when investigating this. We further suggest that optimal immune responses are context specific, rather than generic; that is, a maximum immune response is not necessarily optimal. The nature of an optimal immune response will depend on the specific circumstances and infection status of the animal. Identifying and understanding such optimality requires that the effects of different immune strategies on fitness be considered. PMID- 16701456 TI - The tree of eukaryotes. AB - Recent advances in resolving the tree of eukaryotes are converging on a model composed of a few large hypothetical 'supergroups', each comprising a diversity of primarily microbial eukaryotes (protists, or protozoa and algae). The process of resolving the tree involves the synthesis of many kinds of data, including single-gene trees, multigene analyses, and other kinds of molecular and structural characters. Here, we review the recent progress in assembling the tree of eukaryotes, describing the major evidence for each supergroup, and where gaps in our knowledge remain. We also consider other factors emerging from phylogenetic analyses and comparative genomics, in particular lateral gene transfer, and whether such factors confound our understanding of the eukaryotic tree. PMID- 16701457 TI - Assessing dinosaur growth patterns: a microscopic revolution. AB - Some of the longest standing questions in dinosaur paleontology pertain to their development. Did dinosaurs grow at slow rates similar to extant reptiles or rapidly similar to living birds and mammals? How did some forms attain gigantic proportions? Conversely, how did birds (avian dinosaurs) become miniaturized? New data on dinosaur longevity garnered from bone microstructure (i.e. osteohistology) are making it possible to assess basic life-history parameters of the dinosaurs such as growth rates and timing of developmental events. Analyses of these data in an evolutionary context are enabling the identification of developmental patterns that lead to size changes within the Dinosauria. Furthermore, this rich new database is providing inroads for studying individual and population biology. All in all, paleohistological research is proving to be the most promising avenue towards gaining a comprehensive understanding of dinosaur biology. PMID- 16701458 TI - Ecological consequences of phenotypic plasticity. AB - Phenotypic plasticity is widespread in nature, and often involves ecologically relevant behavioral, physiological, morphological and life-historical traits. As a result, plasticity alters numerous interactions between organisms and their abiotic and biotic environments. Although much work on plasticity has focused on its patterns of expression and evolution, researchers are increasingly interested in understanding how plasticity can affect ecological patterns and processes at various levels. Here, we highlight an expanding body of work that examines how plasticity can affect all levels of ecological organization through effects on demographic parameters, direct and indirect species interactions, such as competition, predation, and coexistence, and ultimately carbon and nutrient cycles. PMID- 16701459 TI - Are plant DNA barcodes a search for the Holy Grail? AB - In a recent study, Kress et al. compared two plant genomes to seek out plant DNA barcodes. Two promising markers balanced the variability that is needed to distinguish species with conserved primer regions that enable universal amplification. Although this study is the most rigorous effort to date, problems from earlier barcoding efforts, such as the use of non-evolutionary species concepts and differential sorting of genes and species, could reemerge. Single gene barcoding might not be universally effective owing to inherent inaccuracies. Kress et al. suggest the use of multiple genes, reflecting an integrated approach that is likely to be the best answer to identifying species quickly and accurately. PMID- 16701460 TI - The ever-flickering light. AB - To date, ecologists involved in global change have focused on the consequences of changes in air temperature. Concurrently, the amount of sunlight reaching the surface of the Earth has been declining, resulting in so-called 'global dimming'. Now, Wild et al. and Pinker et al. have reported a reversal in this trend in some regions that has occurred over the past 15 years or so. These new findings, combined with earlier work, show that the transparency of the atmosphere can vary substantially over periods of at least 20-50 years. Thus, the ecological consequences of sustained trends in the occurrence of sunlight at the surface of the Earth need a more careful assessment than was previously thought. PMID- 16701461 TI - The hitchhiker's guide to avian malaria. AB - The ecological mechanisms underlying the dispersal of parasites are poorly understood, which is of particular concern in view of currently emerging infectious diseases. In a new study, Perez-Tris and Bensch examined the distribution and prevalence of avian malaria in a migratory bird across Western Europe. They concluded that repeated independent evolution of year-round transmission has enabled some avian malaria lineages to become more widespread, and more prevalent, than lineages that are transmitted only during the summer. This study blurs the boundaries of evolutionary ecology, epidemiology and macroecology with great potential for cross-disciplinary research. PMID- 16701462 TI - Ernst Mayr, genetics and speciation. PMID- 16701463 TI - Polyploidization, hybridization and geographical parthenogenesis. PMID- 16701465 TI - What sets the odds of winning and losing? AB - Social experience influences the outcome of conflicts such that winners are more likely to win again and losers will more likely lose again, even against different opponents. Although winner and loser effects prevail throughout the animal kingdom and crucially influence social structures, the ultimate and proximate causes for their existence remain unknown. We propose here that two hypotheses are particularly important among the potential adaptive explanations: the 'social-cue hypothesis', which assumes that victory and defeat leave traces that affect the decisions of subsequent opponents; and the 'self-assessment hypothesis', which assumes that winners and losers gain information about their own relative fighting ability in the population. We discuss potential methodologies for experimental tests of the adaptive nature of winner and loser effects. PMID- 16701466 TI - How do adaptive immune systems control pathogens while avoiding autoimmunity? AB - Immune systems face a daunting control challenge. On the one hand, they need to minimize damage from pathogens, without wasting energy and resources, but on the other must avoid initiating or perpetuating autoimmune responses. Finally, because pathogens interfere with immune function, immune systems must be robust against sabotage. We describe here how these challenges are met by two immune systems, the intracellular RNA interference system and the vertebrate CD8 T-cell response. We extrapolate from these two systems to propose principles for strategically robust control. PMID- 16701467 TI - Uncovering evolutionary patterns of gene expression using microarrays. AB - The advent of microarray technology is providing new insights into fundamental questions in evolutionary biology. Here, we review the recent literature on the use of microarrays to study the evolution of genome-wide patterns of gene expression within and between species. Large levels of variation in gene expression patterns have been observed at the intra and interspecific level, and a substantial fraction of transcriptional variation has a genetic component that is contributed by changes in both cis-acting and trans-acting regulatory elements. We argue that there is solid evidence to show that the temporal dynamics of transcriptional variation is largely determined by natural selection, with the fraction of the transcriptome more closely related to sex and reproduction evolving more rapidly. PMID- 16701468 TI - Conservation physiology. AB - Conservation biologists increasingly face the need to provide legislators, courts and conservation managers with data on causal mechanisms underlying conservation problems such as species decline. To develop and monitor solutions, conservation biologists are progressively using more techniques that are physiological. Here, we review the emerging discipline of conservation physiology and suggest that, for conservation strategies to be successful, it is important to understand the physiological responses of organisms to their changed environment. New physiological techniques can enable a rapid assessment of the causes of conservation problems and the consequences of conservation actions. PMID- 16701469 TI - Do telomere dynamics link lifestyle and lifespan? AB - Identifying and understanding the processes that underlie the observed variation in lifespan within and among species remains one of the central areas of biological research. Questions directed at how, at what rate and why organisms grow old and die link disciplines such as evolutionary ecology to those of cell biology and gerontology. One process now thought to have a key role in ageing is the pattern of erosion of the protective ends of chromosomes, the telomeres. Here, we discuss what is currently known about the factors influencing telomere regulation, and how this relates to fundamental questions about the relationship between lifestyle and lifespan. PMID- 16701471 TI - Kin selection is the key to altruism. AB - Kin selection theory, also known as inclusive fitness theory, has been the subject of much debate and misunderstanding. Nevertheless, the idea that relatedness among individuals can drive the evolution of altruism has emerged as a central paradigm in evolutionary biology. Or has it? In two recent articles, E.O. Wilson argues that kin selection should no longer be considered the main explanation for the evolution of altruism in insect societies. Here, we discuss what these articles say about kin selection and how it relates to the theory. We conclude that kin selection remains the key explanation for the evolution of altruism in eusocial insects. PMID- 16701470 TI - Genetic sex determination and extinction. AB - Genetic factors can affect the probability of extinction either by increasing the effect of detrimental variants or by decreasing the potential for future adaptive responses. In a recent paper, Zayed and Packer demonstrate that low variation at a specific locus, the complementary sex determination (csd) locus in Hymenoptera (ants, bees and wasps), can result in a sharply increased probability of extinction. Their findings illustrate situations in which there is a feedback process between decreased genetic variation at the csd locus owing to genetic drift and decreased population growth, resulting in an extreme type of extinction vortex for these ecologically important organisms. PMID- 16701472 TI - Molecular cryptozoology meets the Sasquatch. PMID- 16701473 TI - Megafaunal extinction: climate, humans and assumptions. PMID- 16701475 TI - Orchid diversity--beyond deception. PMID- 16701477 TI - The value of the IUCN Red List for conservation. AB - The IUCN Red List of Threatened Species is the most comprehensive resource detailing the global conservation status of plants and animals. The 2004 edition represents a milestone in the four-decade long history of the Red List, including the first Global Amphibian Assessment and a near doubling in assessed species since 2000. Moreover, the Red List assessment process itself has developed substantially over the past decade, extending the value of the Red List far beyond the assignation of threat status. We highlight here how the Red List, in conjunction with the comprehensive data compiled to support it and in spite of several important limitations, has become an increasingly powerful tool for conservation planning, management, monitoring and decision making. PMID- 16701478 TI - A user's guide to animal welfare science. AB - Here, I provide a guide for those new to the burgeoning field of animal welfare science as to what this comprehensive, relatively young discipline is all about. Drawing on all branches of biology, including behavioural ecology and neuroscience, the science of animal welfare asks three big questions: Are animals conscious? How can we assess good and bad welfare in animals? How can we use science to improve animal welfare in practice? I also provide guidelines for an evidence-based approach to welfare issues for policy makers and other users of animal welfare research. PMID- 16701479 TI - Sexually selected signals are not similar to sports handicaps. AB - The handicap principle is a simple but powerful metaphor that has had a major impact on how biologists study and understand sexual selection. Here, I show that its application to signalling in sexual selection is not a valid generalization from its roots in economics. Although some signalling systems, with additive costs and benefits, have solutions that resemble sports handicaps, the signalling in sexual selection has multiplicative costs and benefits, and solutions that do not resemble sports handicaps. The sports analogy is technically incorrect, metaphorically misleading and a poor guide for empirical research on the signalling in sexual selection. The evolution of sexually selected signals is not a missing piece of Darwin's puzzle; it is an integral piece of the process of evolution by natural selection, and it should be approached with the same tools that we bring to bear on the evolution of other correlated traits involved in social interactions. PMID- 16701480 TI - Diversity and flexibility of sex-change strategies in animals. AB - Here, we review recent empirical advances that have improved our understanding of why and when sex change occurs. We show that sex-changing animals use a greater diversity of strategies to increase their reproductive success than was previously recognized: some individuals change sex early, others change sex late, some individuals change sex more than once, and others do not change sex at all. These different strategies can be unified by the principle that individuals change sex when it increases their reproductive value. The breeding tactics (male, female or non-breeder) adopted by individuals often appear to be adaptive responses to their own social-ecological context and variation in these conditions results in significant differences in the timing of sex change within and between species. PMID- 16701481 TI - Regional and global concerns over wetlands and water quality. AB - Water quality in many stream catchments and river basins is severely impacted by nutrient enrichment as a result of agriculture. Water-resource managers worldwide are considering the potential role of riparian zones and floodplain wetlands in improving stream-water quality, as there is evidence at the site scale that such wetlands are efficient at removing nutrients from through-flowing water. However, recent studies have highlighted disadvantages of such use of wetlands, including emissions of greenhouse gases and losses of biodiversity that result from prolonged nutrient loading. Here, we discuss the water purification function of wetlands at the site and catchment scale and suggest ways in which these disadvantages could be overcome. PMID- 16701482 TI - Tropical rainforests and the need for cross-continental comparisons. AB - Ecologists have tended to overemphasize the common features of tropical rainforests on different continents. In reality, the five major tropical rainforest regions (tropical America, Africa, Southeast Asia, Madagascar and New Guinea) are distinct ecological and biogeographical entities. Although it is easy to find examples of at least superficial convergence between unrelated organisms in these different regions, there are many other cases where convergence is incomplete or there are no obvious ecological equivalents. Pantropical comparisons with standardized methods are needed for the insights that they can provide into rainforest ecology and the help that they can offer in identifying conservation strategies that are appropriate to regional conditions. Here, we suggest ways in which the practical difficulties of such pantropical comparisons can be minimized. PMID- 16701483 TI - Climate change and the migration capacity of species. AB - In a recent paper, McLachlan et al. presented evidence that migration rates of two tree species at the end of the last glacial (c. 10-20 thousand years ago) were much slower than was previously thought. These results provide an important insight for climate-change impacts studies and suggest that the ability of species to track future climate change is limited. However, the detection of late glacial refugia close to modern range limits also implies that some of our most catastrophic projections might be overstated. PMID- 16701484 TI - Root foraging theory put to the test. AB - Roots have a tremendous plasticity that has long fascinated plant scientists. Root proliferation into enriched soil patches is commonly considered as a way for plants to acquire patchily distributed soil resources. In a recent synthetic study involving the responses of over 100 species, Kembel and Cahill concluded that long-standing generalizations explaining variation in root proliferation should be reconsidered. PMID- 16701485 TI - Changing values of urban biodiversity: a reply to Miller. PMID- 16701487 TI - Towards the global monitoring of biodiversity change. AB - Governments have set the ambitious target of reducing biodiversity loss by the year 2010. The scientific community now faces the challenge of assessing the progress made towards this target and beyond. Here, we review current monitoring efforts and propose a global biodiversity monitoring network to complement and enhance these efforts. The network would develop a global sampling programme for indicator taxa (we suggest birds and vascular plants) and would integrate regional sampling programmes for taxa that are locally relevant to the monitoring of biodiversity change. The network would also promote the development of comparable maps of global land cover at regular time intervals. The extent and condition of specific habitat types, such as wetlands and coral reefs, would be monitored based on regional programmes. The data would then be integrated with other environmental and socioeconomic indicators to design responses to reduce biodiversity loss. PMID- 16701488 TI - A general eco-evolutionary framework for understanding bioinvasions. AB - Studies of bioinvasions have revealed various strategies of invasion, depending on the ecosystem invaded and the alien species concerned. Here, we consider how migration (as a demographic factor), as well as ecological and evolutionary changes, affect invasion success. We propose three main theoretical scenarios that depend on how these factors generate the match between an invader and its new environment. Our framework highlights the features that are common to, or differ among, observed invasion cases, and clarifies some general trends that have been previously highlighted in bioinvasions. We also suggest some new directions of research, such as the assessment of the time sequence of demographic, genetic and environmental changes, using detailed temporal surveys. PMID- 16701489 TI - Metabolic theories in ecology. AB - The Metabolic Theory of Ecology (MTE) and the earlier theory of Dynamic Energy Budgets (DEB) are both founded on simple mechanistic descriptions of how individual organisms take up and use energy and material. Such descriptions should enable predictions to be made of pools and flows of energy and matter in populations, communities and ecosystems. MTE builds on the idea that the transport of resources through a fractal-like branching network causes the supply rate to cells to scale as a (3/4) power of body mass, whereas DEB is based on the concept that rates of basic physiological processes are proportional to surface area or to body volume. Here, I critically compare both theories, highlight similarities and inconsistencies, and show where the approaches deviate. I argue that, to arrive at a sound theoretical basis of the energy budget of individual organisms, both views should be tested fully, but only after inconsistencies have been addressed. PMID- 16701490 TI - Demography in an increasingly variable world. AB - Recent advances in stochastic demography provide unique insights into the probable effects of increasing environmental variability on population dynamics, and these insights can be substantially different compared with those from deterministic models. Stochastic variation in structured population models influences estimates of population growth rate, persistence and resilience, which ultimately can alter community composition, species interactions, distributions and harvesting. Here, we discuss how understanding these demographic consequences of environmental variation will have applications for anticipating changes in populations resulting from anthropogenic activities that affect the variance in vital rates. We also highlight new tools for anticipating the consequences of the magnitude and temporal patterning of environmental variability. PMID- 16701491 TI - The evolution of echolocation in bats. AB - Recent molecular phylogenies have changed our perspective on the evolution of echolocation in bats. These phylogenies suggest that certain bats with sophisticated echolocation (e.g. horseshoe bats) share a common ancestry with non echolocating bats (e.g. Old World fruit bats). One interpretation of these trees presumes that laryngeal echolocation (calls produced in the larynx) probably evolved in the ancestor of all extant bats. Echolocation might have subsequently been lost in Old World fruit bats, only to evolve secondarily (by tongue clicking) in this family. Remarkable acoustic features such as Doppler shift compensation, whispering echolocation and nasal emission of sound each show multiple convergent origins in bats. The extensive adaptive radiation in echolocation call design is shaped largely by ecology, showing how perceptual challenges imposed by the environment can often override phylogenetic constraints. PMID- 16701492 TI - Extinct mammalian biodiversity of the ancient New World tropics. AB - The origins of mammalian biodiversity in the New World tropics extend back >25 million years, represented by clades that were originally endemic to South America, North America or Africa. Since then, these mammalian clades have been greatly affected by climatic, physiographic and biological changes. The Isthmian land bridge, which formed approximately 4 million years ago between North and South America, resulted in the maximum diversity of 17 New World tropical mammalian orders during the Great American Interchange. This diversity was subsequently reduced to 12 orders as a result of competition, climate change and human impacts. Here, I discuss how the fossil record is now providing a rich archive of past biodiversity, presenting unique evidence of the origins, macroevolution, macro-ecology and extinction of New World tropical mammals. PMID- 16701493 TI - Viability of vaginal probiotic lactobacilli during refrigerated and frozen storage. AB - The viability of six different strains of probiotic vaginal Lactobacillus was examined in two different cryoprotective media, during refrigerated versus frozen storage, and using two traditional types of stock cultures for starting the biomass production. Freezing at -20 degrees C and -70 degrees C had much less adverse effect on viability than did storage at 7 degrees C, and the reduction in viability was greater at -20 degrees C than at -70 degrees C. The strains showed variation in the extent of the viability losses during both types of storage. Milk-yeast extract (MYE) was shown to be the more suitable protective medium to maintain viability of the strains during the storage. The vaginal Lactobacillus strains are most stable in MYE at -70 degrees C with only a small decrease of the viability observed under these conditions. The viable cell counts of Lactobacillus paracasei CRL 1251 and CRL 1289, L. crispatus CRL 1266 and L. salivarius CRL 1328 remained around 1 x 10(8) CFU/mL after 24 months of storage at -70 degrees C, or up to 18 months for L. acidophilus CRL 1259. PMID- 16701494 TI - Interference of thiosulfate during colorimetric analysis of hexavalent chromium using 1,5-diphenylcarbazide method. AB - Thiosulfate (S(2)O(3)(2-)) contained in the media for autotrophic Cr (VI) reduction was found to interfere with Cr (VI) measurement following the 1,5 diphenylcarbazide (DPC) method. The interference was confirmed at several abiotic and biotic conditions, and was influenced by S(2)O(3)(2-) concentration, pH, and the media components. At neutral to alkaline pH, 500 mg/L of S(2)O(3)(2-) did not cause interference, while 4 mg/L of S(2)O(3)(2-) resulted in the interference at pH 2.0. Atomic absorption spectrophotometry could be an alternative method when the interference by S(2)O(3)(2-) is expected. PMID- 16701495 TI - Effect of ammonia on the methanogenic activity of methylaminotrophic methane producing Archaea enriched biofilm. AB - Ammonia is a metabolic product in the decomposition of protein wastes, and has a recognized inhibitory effect on methanogenesis; this effect has been slightly quantified on methanogenic biofilms and particularly those populated by methanogenic Archaea which produce ammonia as a catabolic product from methylated amines. This paper presents studies on the effect of ammonia on maximum methanogenic activity of anaerobic biofilms enriched by methylaminotrophic methane producing Archaea (mMPA). The effect of unionized free ammonia on the specific maximum methanogenic activity of a mMPA enriched biofilm was studied, using 250 mL flasks containing ceramic rings colonized by 30 day-old experimental biofilm and adding 48.8 (control system), 73.8, 98.8, 148.8, 248.8, 448.8 and 848.8 mg NH(3)-N/L. The systems were maintained for ten days at a pH of 7.5 and temperature of 37 degrees C. The results showed that at 848.8 mg NH(3)-N/L, biofilm methane production required 36 h adaptation period, prior to entering into maximum production phase. The highest maximum methanogenic activity reached a value of 2.337+/-0.213 g COD methane/g VSS *day when 48.8 mg NH(3)-N/L was added, and inhibition was clearly observed in those systems above 148.8 mg NH(3) N/L, producing under 1.658+/-0.185 g COD methane/g VSS *day. The lowest methanogenic activity reached was 0.639+/-0.162 g COD methane/g VSS *day at the system added with 848.8 mg NH(3)-N/L. When applying the Luong and non-competitive inhibition models, the best fit was obtained with the non-competitive model, which predicted 50% inhibition of methanogenic activity at 365.288 mg NH(3)-N/L. PMID- 16701496 TI - Effects of different digestible carbohydrates on bile acid metabolism and SCFA production by human gut micro-flora grown in an in vitro semi-continuous culture. AB - The main source of carbon in the human large intestine comes from carbohydrates like starches and oligosaccharides which remain unchanged by gastric digestion. These polysaccharides are metabolised in the colon by saccharolytic bacteria whose composition is dependent upon the substrate availability. Among the metabolites produced, the short-chain fatty acids (SCFA) are important for colon function and to prevent diseases. In particular, butyrate affects several cellular functions (proliferation, membrane synthesis, sodium absorption), and it has been shown to be protective against colorectal cancer. In addition, faecal bacteria are responsible for the conversion of primary bile acids (BA) to secondary BA, which are considered tumor promoters. In this study we investigated the in vitro effect of different substrates (CrystaLean starch, xylo oligosaccharides, corn starch) supplied to human faecal micro-flora, on the SCFA production, on the bowel micro-flora composition and on the primary BA conversion rate. In addition, with corn starch as substrate, we considered the effect of enriching normal human faecal micro-flora with lactobacilli and bifidobacteria, on the above reported parameters. PMID- 16701497 TI - Glucose toxicity effect and accumulation of methylglyoxal by the periodontal anaerobe Bacteroides forsythus. AB - Growth of the periodontal pathogen Bacteroides forsythus in broth cultures showed inhibition in the presence of 10mM glucose added to the medium. Glucose inhibition in a number of rumen bacteria has been attributed to the accumulation of methylglyoxal (MG), a highly reactive electrophile known to exhibit cytotoxic effects. HPLC analysis revealed elevated concentrations of MG in cultures of seven strains of B. forsythus. MG rose during growth to a maximum at the time of growth inhibition. Maximum MG concentrations for strain ATCC 43037 were 60.6+/ 8.2 microM without added glucose, and 185.5+/-21.5 microM (P<0.014) with 10mM added glucose. Other strains gave values ranging from 24-91 microM and 100-326 microM MG, respectively. Both free and reversibly bound MG were detected in the bacterial cells and in the cell-free culture fluid. Disk sensitivity tests indicated that three B. forsythus strains exhibited different sensitivities to growth inhibition by added MG. Altogether, the results demonstrated the production and accumulation by B. forsythus of high levels of MG in vitro. MG accumulation appears to be related to the marked auto-inhibitory glucose-toxicity effect observed with B. forsythus strains, an effect that must be considered in the design of optimal media for the culture of this fastidious species. In the diseased periodontal pocket, production of the highly reactive, cytotoxic MG by B. forsythus may contribute significantly to disease pathogenesis. PMID- 16701498 TI - Isolation, growth on prebiotics and probiotic potential of novel bifidobacteria from pigs. AB - Bifidobacteria were isolated from the faeces of pigs of various ages and examined for their potential use as probiotics in combination with di- and oligosaccharides. Ninty-six per cent of the isolates were found to have characteristics in common with Bifidobacterium boum, B. thermophilum and B. choerinum. B. thermophilum was most commonly isolated from sows, whereas most of the other strains were isolated from piglets. A few strains of each species were able to grow in the presence of air. A microplate assay was modified to allow comparison of growth on different substrates. Di- and oligosaccharides considered to promote bifidobacteria were screened for their ability to support growth of selected isolates in vitro. Growth on these substrates varied within and between species. Of the fructose oligosaccharides tested, Actilight P supported the best growth of the widest range of strains. The strains which grew best on the disaccharide lactulose were related to B. choerinum and some of these strains grew on xylo-oligosaccharides. It seems that prebiotic di- and oligosaccharides may have both a species and intra-species/strain selective effect. B. choerinum appeared to be well adapted to the gut of pre-weaned piglets. PMID- 16701499 TI - Anaerofustis stercorihominis gen. nov., sp. nov., from human feces. AB - Phenotypic and phylogenetic studies were performed on an unidentified Gram positive, strictly anaerobic, non-spore-forming, rod-shaped bacterium isolated from human feces. The organism was catalase-negative, resistant to 20% bile, produced acetic and butyric acids as end products of glucose metabolism, and possessed a G+C content of approximately 70 mol%. Comparative 16S rRNA gene sequencing demonstrated that the unidentified bacterium was a member of the Clostridium sub-phylum of the Gram-positive bacteria, and formed a loose association with rRNA cluster XV. Sequence divergence values of 12% or greater were observed between the unidentified bacterium and all other recognized species within this and related rRNA clusters. Treeing analysis showed the unknown anaerobe formed a deep line branching near to the base of rRNA cluster XV and phylogenetically represents a hitherto unknown taxon, distinct from Acetobacterium, Eubacterium sensu stricto, Pseudoramibacter and other related organisms. Based on both phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium from feces be classified in a new genus Anaerofustis, as Anaerofustis stercorihominis sp. nov. The type strain of Anaerofustis stercorihominis is ATCC BAA-858(T)=CCUG 47767(T). PMID- 16701500 TI - Antimicrobial susceptibility of Clostridium difficile isolated from neonatal pigs with enteritis. AB - The minimum inhibitory concentration (MIC) of eight antimicrobial agents was determined by the agar dilution method for 80 isolates of Clostridium difficile from neonatal pigs with enteritis. MICs(50) for erythromycin, tilmicosin, and tylosin were relatively low (0.25-0.50 microg/mL), but MICs(90) (64 or > or =256 microg/mL) suggest in vivo resistance of a proportion of isolates. Susceptibility to tetracycline varied widely, with MIC(50) and MIC(90) of 8 and 32 microg/mL, respectively. The MICs(90) for tiamulin (8 microg/mL) and virginiamycin (16 microg/mL) suggest moderate susceptibility. Bacitracin and ceftiofur (MICs(90) > or =256 microg/mL) have little activity against C. difficile. Tiamulin and virginiamycin may decrease fecal shedding of C. difficile by sows, and erythromycin, tetracycline, and tylosin may be useful for treatment of infected piglets. PMID- 16701501 TI - Insights in metabolism and toxin production from the complete genome sequence of Clostridium tetani. AB - The decryption of prokaryotic genome sequences progresses rapidly and provides the scientific community with an enormous amount of information. Clostridial genome sequencing projects have been finished only recently, starting with the genome of the solvent-producing Clostridium acetobutylicum in 2001. A lot of attention has been devoted to the genomes of pathogenic clostridia. In 2002, the genome sequence of C. perfringens, the causative agent of gas gangrene, has been released. Currently in the finishing stage and prior to publication are the genomes of the foodborne botulism-causing C. botulinum and of C. difficile, the causative agent of a wide spectrum of clinical manifestations such as antibiotic associated diarrhea. Our team sequenced the genome of neuropathogenic C. tetani, a Gram-positive spore-forming bacterium predominantly found in the soil. In deep wound infections it occasionally causes spastic paralysis in humans and vertebrate animals, known as tetanus disease, by the secretion of potent neurotoxin, designated tetanus toxin. The toxin blocks the release of neurotransmitters from presynaptic membranes of interneurons of the spinal cord and the brainstem, thus preventing muscle relaxation. Fortunately, this disease is successfully controlled through immunization with tetanus toxoid, a formaldehyde-treated tetanus toxin, but nevertheless, an estimated 400,000 cases still occur each year, mainly of neonatal tetanus. The World Health Organization has stated that neonatal tetanus is the second leading cause of death from vaccine preventable diseases among children worldwide. This minireview focuses on an analysis of the genome sequence of C. tetani E88, a vaccine production strain, which is a toxigenic non-sporulating variant of strain Massachusetts. The genome consists of a 2,799,250 bp chromosome encoding 2618 open reading frames. The tetanus toxin is encoded on a 74,082 kb plasmid, containing 61 genes. Additional virulence-related factors as well as an insight into the metabolic strategy of C. tetani with regard to its pathogenic phenotype will be presented. The information from other clostridial genomes by means of comparative analysis will also be explored. PMID- 16701502 TI - Initiation of endospore formation in Clostridium acetobutylicum. AB - Endospore formation in bacilli and clostridia shows remarkable similarities in morphology as well as in physiological and molecular biological cellular events. Major differences are the formation of clostridial stage cells and granulose accumulation in clostridia. In both genera, a cascade of sigma factors is activated after septation (by help of sigma(H) and Spo0A approximately P) in the sequence sigma(F), sigma(E), sigma(G), and sigma(K). Of these, sigma(F) and sigma(G) are active inside the forespore and are regulated by anti-sigma factors and anti-anti-sigma factors, whereas sigma(E) and sigma(K) (mother cell-specific sigma factors) are synthesized as precursor proteins and activated by proteolysis. Each of these sigma factors allows transcription of a specific set of genes and operons, thus leading to the orchestral expression of stage-specific proteins required for successful sporulation. Both, the genetic organization of the respective operons and the expression pattern of the sigma factors are very similar in Bacillus subtilis and Clostridium acetobutylicum, the model organisms of the two genera. However, a major regulatory difference is found in initiation of endospore formation. Genome sequencing revealed that clostridia do not contain components of the so-called phosphorelay, with the exception of the essential transcription factor Spo0A. This might reflect recognition of different environmental signals, as for clostridia nutrient limitation is no prerequisite for sporulation. In contrast to Bacillus, the clostridial sigH gene is constitutively expressed at a low level, with no increase at the onset of spore formation. The spo0A gene in C. acetobutylicum is also constitutively expressed, but Spo0A synthesis only occurs during the early and mid-exponential growth phase, indicating a posttranscriptional or cotranslational regulation. Mutational studies revealed an important regulatory function of a dual palindrome region upstream of the spo0A gene of C. acetobutylicum, part of which overlaps with a Spo0A-binding site. In addition to controlling sporulation genes, phosphorylated clostridial Spo0A is involved in regulation of acetone and butanol synthesis. PMID- 16701503 TI - The development of Clostridium difficile genetic systems. AB - Clostridum difficile is a major cause of healthcare-associated disease in the western world, and is particularly prominent in the elderly. Its incidence is rising concomitant with increasing longevity. More effective countermeasures are required. However, the pathogenesis of C. difficile infection is poorly understood. The lack of effective genetic tools is a principal reason for this ignorance. For many years, the only tools available for the transfer of genes into C. difficile have been conjugative transposons, such as Tn916, delivered via filter mating from Bacillus subtilis donors. They insert into a preferred site within the genome. Therefore, they may not be employed for classical mutagenesis studies, but can be employed to modulate gene function through the delivery of antisense RNA. Attempts to develop transformation procedures have so far met with little success. However, in recent years the situation has been dramatically improved through the demonstration of efficient conjugative transfer of both replication-proficient and replication-deficient plasmids from Escherichia coli donors. This efficient transfer can only be achieved in certain strains through negation of the indigenous restriction barrier, and is generally most effective when the plasmid employed is based on the replicon of the C. difficile plasmid, pCD6. PMID- 16701504 TI - The IStron CdISt1 of Clostridium difficile: molecular symbiosis of a group I intron and an insertion element. AB - The IStron CdISt1 was first discovered as an insertion into the tcdA gene of the clinical isolate C34. It combines structural and functional properties of a group I intron at its 5'-end with those of an insertion element at its 3'-end. Up to date four different types could be found, mainly differing in their IS-element portions. Contrasting classical group I introns, CdISt1 is always integrated in ORFs encoding bacterial protein. In case CdISt1 had only the IS-element function such insertion would inactivate the protein encoded by the host gene. It is only due to the self-splicing activity of the group I intron parts that CdISt1 integration does not abolish protein function. Both elements seem to exist in molecular symbiosis and CdISt1 could thus be a prototype of a novel class of genetic elements. Moreover, integration of the CdISt1 into the genome could be advantageous for the bacterium, a motor function for evolution of bacterial proteins is discussed. In clinical practice CdISt1 might well serve as a tool for epidemiological studies of C. difficile infections. PMID- 16701505 TI - Organization and regulation of the neurotoxin genes in Clostridium botulinum and Clostridium tetani. AB - Botulinum and tetanus neurotoxins are structurally and functionally related 150 kDa proteins that are potent inhibitors of neuroexocytosis. Botulinum neurotoxin associates with non-toxic proteins to form complexes of various sizes. The botulinum neurotoxin and non-toxic protein genes are clustered in a DNA segment called the botulinum locus. This locus is probably located on a mobile or degenerate mobile element, which accounts for the various genomic localizations (chromosome, plasmid, phage) in different Clostridium botulinum types. The botulinum neurotoxin and non-toxic protein genes are organized in two polycistronic operons (ntnh-bont and ha operons) transcribed in opposite orientations. The gene that separates the two operons of the botulinum locus in C. botulinum A encodes a 21 kDa protein BotR/A, which is a positive regulator of the expression of the botulinum locus genes. Similarly, in Clostridium tetani, the gene located immediately upstream of the tetanus toxin gene, encodes a positive regulatory protein, TetR. BotR and TetR are possibly alternative sigma factors related to TxeR and UviA, which regulate C. difficile toxin and C. perfringens bacteriocin production, respectively. TxeR and UviA define a new sub group of the sigma(70) family of RNA polymerase initiation factors. In addition, the C. botulinum genome contains predicted two-component system genes, some of which are possibly involved in regulation of toxinogenesis. PMID- 16701506 TI - The actin-ADP-ribosylating Clostridium botulinum C2 toxin. AB - Clostridium botulinum C2 toxin is the prototype of actin-ADP-ribosylating toxins. The toxin consists of the enzyme component C2I and the separated binding/translocation component C2II. C2II is proteolytically activated to form heptamers, which bind the enzyme component. After endocytosis of the receptor toxin complex, the enzyme component enters the cytosol from an acidic endosomal compartment to modify G-actin at arginine177. Recent data indicate that chaperons are involved in the translocation process of the toxin. PMID- 16701507 TI - New insights into the cytotoxic mechanisms of Clostridium perfringens enterotoxin. AB - Clostridium perfringens type A isolates producing the 35 kDa enterotoxin (CPE) are an important cause of food poisoning, human non-foodborne gastrointestinal (GI) disease, and some veterinary GI diseases. Studies using CPE knock-out mutants confirmed the importance of enterotoxin expression for the enteric virulence of CPE-positive type A isolates. CPE action involves formation of a series of complexes in mammalian plasma membranes. One such CPE-containing complex (of approximately 155 kDa) is important for the induction of plasma membrane permeability alterations, which are responsible for killing enterotoxin treated mammalian cells. Those membrane permeability changes damage the epithelium, allowing the enterotoxin to interact with the tight junction (TJ) protein occludin. CPE:occludin interactions result in formation of an approximately 200 kDa CPE complex and internalization of occludin into the cytoplasm. That removal of occludin (and possibly other proteins) damages TJs and disrupts the normal paracellular permeability barrier of the intestinal epithelium, which may contribute to CPE-induced diarrhea. Recent studies demonstrated that low CPE doses kill mammalian cells by inducing a classic apoptotic pathway involving mitochondrial membrane depolarization, cytochrome C release, and caspase 3/7 activation. In contrast, high enterotoxin doses induce oncosis, a proinflammatory event. Thus, inflammation may also contribute to the GI symptoms of patients whose intestines contain high CPE levels. In summary, CPE is a unique, multifunctional toxin with cytotoxic, TJ-damaging, and (probably) proinflammatory activities. PMID- 16701508 TI - Effects of Clostridium perfringens phospholipase C in mammalian cells. AB - Clostridium perfringens phospholipase C (Cp-PLC), the major virulence factor in the pathogenesis of gas gangrene, is a Zn(2+) metalloenzyme with lecithinase and sphingomyelinase activities. Its structure shows an N-terminal domain containing the active site, and a C-terminal Ca(2+) binding domain required for membrane interaction. Although the knowledge of the structure of Cp-PLC and its interaction with aggregated phospholipids has advanced significantly, an understanding of the effects of Cp-PLC in mammalian cells is still incomplete. Cp PLC binds to artificial bilayers containing cholesterol and sphingomyelin or phosphatidylcholine (PC) and degrades them, but glycoconjugates present in biological membranes influence its binding or positioning toward its substrates. Studies with Cp-PLC variants harboring single amino-acid substitutions have revealed that the active site, the Ca(2+) binding region, and the membrane interacting surface are required for cytotoxic and haemolytic activity. Cp-PLC causes plasma membrane disruption at high concentrations, whereas at low concentrations it perturbs phospholipid metabolism, induces DAG generation, PKC activation, Ca(2+) mobilization, and activates arachidonic acid metabolism. The cellular susceptibility to Cp-PLC depends on the composition of the plasma membrane and the capacity to up-regulate PC synthesis. The composition of the plasma membrane determines whether Cp-PLC can bind and acquire its active conformation, and thus the extent of phospholipid degradation. The capacity of PC synthesis and the availability of precursors determine whether the cell can replace the degraded phospholipids. Whether the perturbations of signal transduction processes caused by Cp-PLC play a role in cytotoxicity is not clear. However, these perturbations in endothelial cells, platelets and neutrophils lead to the uncontrolled production of intercellular mediators and adhesion molecules, which inhibits bacterial clearance and induces thrombotic events, thus favouring bacterial growth and spread in the host tissues. PMID- 16701509 TI - Perfringolysin O, a cholesterol-binding cytolysin, as a probe for lipid rafts. AB - Gaining an understanding of the structural and functional roles of cholesterol in membrane lipid rafts is a critical issue in studies on cellular signaling and because of the possible involvement of lipid rafts in various diseases. We have focused on the potential of perfringolysin O (theta-toxin), a cholesterol-binding cytolysin produced by Clostridium perfringens, as a probe for studies on membrane cholesterol. We prepared a protease-nicked and biotinylated derivative of perfringolysin O (BCtheta) that binds selectively to cholesterol in cholesterol rich microdomains of cell membranes without causing membrane lesions. Since the domains fulfill the criteria of lipid rafts, BCtheta can be used to detect cholesterol-rich lipid rafts. This is in marked contrast to filipin, another cholesterol-binding reagent, which binds indiscriminately to cell cholesterol. Using BCtheta, we are now searching for molecules that localize specifically in cholesterol-rich lipid rafts. Recently, we demonstrated that the C-terminal domain of perfringolysin O, domain 4 (D4), possesses the same binding characteristics as BCtheta. BIAcore analysis showed that D4 binds specifically to cholesterol with the same binding affinity as the full-size toxin. Cell-bound D4 is recovered predominantly from detergent-insoluble, low-density membrane fractions where raft markers, such as cholesterol, flotillin and Src family kinases, are enriched, indicating that D4 also binds selectively to lipid rafts. Furthermore, a green fluorescent protein-D4 fusion protein (GFP-D4) was revealed to be useful for real-time monitoring of cholesterol in lipid rafts in the plasma membrane. In addition, the expression of GFP-D4 in the cytoplasm might allow the investigations of intracellular trafficking of lipid rafts. The simultaneous visualization of lipid rafts in plasma membranes and inside cells might help in gaining a total understanding of the dynamic behavior of lipid rafts. PMID- 16701510 TI - Diagnosis of Clostridium perfringens intestinal infections in sheep and goats. AB - Clostridium perfringens produces disease in sheep, goats and other animal species, most of which are generically called enterotoxemias. This micro-organism can be a normal inhabitant of the intestine of most animal species including humans, but when the intestinal environment is altered by sudden changes in diet or other factors, C. perfringens proliferates in large numbers and produces several potent toxins that are absorbed into the general circulation or act locally with usually devastating effects on the host. History, clinical signs and gross post-mortem findings are useful tools for establishing a presumptive diagnosis of enterotoxaemia by C. perfringens in sheep and goats, although no definitive diagnosis of these diseases can be made without laboratory confirmation. Because all types of C. perfringens can be normal inhabitants of the intestine of most animals, culture of this micro-organism from intestinal contents of animals has no diagnostic value unless a colony count is performed and large numbers (usually more than 10(4)-10(7)CFU/g) of C. perfringens are found. The most accepted criterion in establishing a definitive diagnosis of enterotoxaemia by C. perfringens is the detection of its toxins in intestinal contents. However, some of the major toxins of C. perfringens (i.e. epsilon toxin) can also be found, albeit in small amounts, in the small intestine of clinically normal sheep, and this poses a diagnostic challenge. In such cases the histopathology of the brain must be used as an alternative diagnostic tool, since the lesions produced by epsilon toxin in the brains of sheep and goats are unique and pathognomonic for C. perfringens type D enterotoxaemia. Ancillary tests, such as measurement of urine glucose or observation of Gram stained smears of intestinal mucosa can be used and, although they have a presumptive diagnostic value when positive, they cannot be used to rule out a diagnosis of enterotoxaemia if they are negative. In conclusion, the diagnosis of C. perfringens infections in animals is complex and it is appropriate to rely on a combination of diagnostic techniques rather than one singe test. PMID- 16701511 TI - Neurological disorders produced by Clostridium perfringens type D epsilon toxin. AB - Although the epsilon toxin of Clostridium perfringens type D produces disease in many species of domestic livestock, neurological disturbance is more common and better studied in sheep. High levels of circulating toxin, especially in lambs, cause cerebral microvascular endothelial damage with disruption of the blood brain barrier leading to a severe, diffuse vasogenic oedema and an acute or peracute clinical course to death. With lower toxin levels, or in partially immune sheep, a focal, bilaterally symmetrical encephalomalacia sometimes occurs in selectively vulnerable brain regions after a more protracted clinical course, but the pathogenesis is uncertain. PMID- 16701512 TI - Lemierre's syndrome: case report in a pediatric patient. AB - Lemierre's syndrome was described in 1936 as a severe oropharyngeal infection followed by septic thrombophlebitis of the internal jugular vein and disseminated metastatic infections. Cases occur typically in previously healthy young adults and children. Fusobacterium necrophorum is the main anaerobic bacterium implicated. We present a septic 2-month-old infant with mastoiditis, multiple sites of osteoarthritis and multiple subcutaneous abscesses. No underlying anatomic or immunologic abnormalities were identified. Fusobacterium necrophorum was recovered from blood and bone samples obtained intraoperatively. Treatment included anaerobic coverage and drainage of septic foci. The patient was discharged home on 35th hospital day with oral amoxicillin-clavulanic acid and he recovered without sequelae. This was the first case of Lemierre's syndrome in our hospital. We want to highlight the absence of jugular vein thrombophlebitis, the presence of mastoiditis as previous infection and the surprising appearance of this infection in an edentulous 2-month-old infant. PMID- 16701514 TI - An in vitro assessment of the effects of broad-spectrum antibiotics on the human gut microflora and concomitant isolation of a Lactobacillus plantarum with anti Candida activities. AB - Chemostat culture was used to determine the effects of the antimicrobial agents tetracycline and nystatin on predominant components of the human gut microflora. Their addition to mixed culture systems caused a non-specific, and variable, decrease in microbial populations, although tetracycline allowed an increase in numbers of yeasts. Both had a profound inhibitory effect upon populations seen as important for gut health (bifidobacteria, lactobacilli). However, a tetracycline resistant Lactobacillus was enriched from the experiments. A combination of genotypic and phenotypic characterisation confirmed its identity as Lactobacillus plantarum. This strain exerted powerful inhibitory effects against Candida albicans. Because of its ability to resist the effects of tetracycline, this organism may be useful as a probiotic for the improved management of yeast related conditions such as thrush and irritable bowel syndrome. PMID- 16701513 TI - Antibiotic-resistant Propionibacterium acnes on the skin of patients with moderate to severe acne in Stockholm. AB - The objective was to study the prevalence and antibiotic susceptibility patterns of Propionibacterium acnes strains isolated from patients with moderate to severe acne in Stockholm, Sweden and to determine the diversity of pulsed-field gel electrophoresis types among resistant P. acnes strains. One hundred antibiotic treated patients and 30 non-antibiotic-treated patients with moderate to severe acne participated in the investigation. Facial, neck and trunk skin samples were taken with the agar gel technique. The susceptibility of P. acnes strains to tetracycline, erythromycin, clindamycin and trimethoprim-sulfamethoxazole was determined by the agar dilution method. The genomic profiles of the resistant strains were determined by pulsed-field gel electrophoresis. In the group of patients treated with antibiotics, resistant P. acnes strains were recovered in 37%, while in the non-antibiotic group of patients the incidence of resistant strains was 13%. Thus antibiotic-resistant P. acnes strains were significantly more often isolated from antibiotic-treated patients with moderate to severe acne than from non-antibiotic-treated patients (odds ratio, 3.8; P=0.01). There was a genetic diversity among the P. acnes strains. Forty-four different patterns of SpeI DNA digests were detected and two predominant clones were found. P. acnes strains exhibited different antibiotic susceptibility patterns and identical genotypes or vice versa. A person can be colonized with different strains with varying degrees of antibiotic resistance. The risk of increased resistance of P. acnes must be considered when treating acne patients with antibiotics, and especially long-term therapy should be avoided. PMID- 16701515 TI - Cytotoxicity and antimicrobial susceptibility of Clostridium difficile isolated from hospitalized children with acute diarrhea. AB - Clostridium difficile is an important pathogen associated with outbreaks of pseudomembranous colitis and other intestinal disorders such as diarrhea. In this study, 181 stool samples from children with and without acute diarrhea were analysed. Eighteen children with acute diarrhea were positive to C. ramosum, C. difficile, C. limosum, C. clostridioforme, C. septicum, C. butyricum, C. innocuum and Clostridium sp. Nineteen children without diarrhea harbored C. ramosum, C. septicum, C. barattii, C. butyricum, C. innocuum, C. sphenoides, C. bifermentans, C. clostridioforme and C. paraputrificum. No patient with diarrhea harbored C. barattii, C. bifermentans, C. paraputrificum and C. sphenoides. In addition, ten C. difficile strains were detected in 5 (5.5%) of the children with diarrhea. Also, no children from control group harbored C. difficile, C. limosum and Clostridium sp. Most of the tested strains were resistant to all the used antimicrobial. Nine C. difficile were toxigenic on VERO cells and by multiplex PCR, six strains showed both toxin A and B genes and three strains showed only toxin B gene. In this study, the presence of C. difficile was not significant, and it is suggested the need of more studies to evaluate the role of clostridia or C. difficile play in the childhood diarrhea and these organisms must be looked for routinely and a periodic evaluation of antimicrobial susceptibility should be performed. PMID- 16701516 TI - Clostridium bartlettii sp. nov., isolated from human faeces. AB - Seven obligately anaerobic, Gram-positive, rod-shaped, spore-forming organisms isolated from human faecal specimens were characterized using phenotypic and molecular taxonomic methods. Strains of the unidentified bacterium used carbohydrates as fermentable substrates, producing acetic acid, isovaleric acid and phenylacetic acid (PAA) as the major products of glucose metabolism, and possessed a G +C content of approximately 29.8 mol%. Comparative 16S rRNA gene sequencing showed that the 7 strains were genetically highly related to each other (displaying >99.5% sequence similarity) and represent a previously unknown sub-line within the Clostridium Cluster XI. The closest described species to the novel bacterium is Clostridium glycolicum, although a 16S rRNA sequence divergence of 4% demonstrates that they represent different species. Genomic DNA DNA pairing studies confirmed the separateness of the unknown species and C. glycolicum (30.6% similarity between the proposed type strain of the novel species, WAL 16138, and C. glycolicum ATCC 14880(T)). Based on morphologic, phenotypic and phylogenetic evidence, it is therefore proposed that the unknown bacterium be classified as C. bartlettii sp. nov. The type strain of C. bartlettii is WAL 16138(T) (= ATCCBAA-827(T)=CCUG48940(T)). PMID- 16701517 TI - Clostridium tunisiense sp. nov., a new proteolytic, sulfur-reducing bacterium isolated from an olive mill wastewater contaminated by phosphogypse. AB - A new sporulated fermentative bacterium designated strain E1(T) (T=type strain), was isolated from an anaerobic mud of an olive mill wastewater basin contaminated by phosphogypse produced by a Tunisian factory. Strain E1(T) was a motile Gram positive slightly curved rod with spherical terminal spore swelling the cell. It grew between 18 degrees C and 43 degrees C with an optimum at 37 degrees C and pH 7.8 (range 5.5-8.7), without NaCl (range 0-3%). Strain E1(T) was a chemoorganotrophic anaerobic bacterium fermenting only proteins and amino acids. Yeast extract was required for growth. Elemental sulfur was used as terminal electron acceptor. The G+C content of the DNA was 32.6 mol%. The closest phylogenetical relatives of strain E1(T) were Clostridium thiosulfatireducens and C. subterminale (97.3% similarity for partial rRNA gene sequences). DNA-DNA hybridization values between strain E1(T) and both species were 17% and 20.8%, respectively. On the basis of differences in genotypic and phenotypic characteristics, strain E1(T) (DSM 15206(T), CIP 107666(T)) is proposed as the type strain of a new species, C. tunisiense sp. nov. GenBank accession number for the 16S rRNA gene sequence of strain E1(T) is AY187622. PMID- 16701518 TI - Atopostipes suicloacale gen. nov., sp. nov., isolated from an underground swine manure storage pit. AB - Phenotypic and molecular genetic studies were performed on an unknown facultative anaerobic, catalase-negative, non-spore-forming, rod-shaped bacterium isolated from a pig manure storage pit. The unknown bacterium was nutritionally fastidious with growth enhanced by the addition of rumen fluid and was phenotypically initially identified as an Eubacterium species. Comparative 16S rRNA gene sequencing studies, however, revealed that the unknown bacterium was phylogenetically distant from Eubacterium limosum (the type species of the genus Eubacterium) and related organisms. Phylogenetically, the unknown species displayed a close association with an uncultured organism from human subgingival plaque and formed an unknown sub-line within a cluster of organisms which includes Alloioccoccus otitis, Alkalibacterium olivoapovliticus, Allofustis seminis, Dolosigranulum pigrum, and related organisms, within the low mol% G+C Gram-positive bacteria. Sequence divergence values of >8% with all known taxonomically recognised taxa, however, clearly indicates the novel bacterium represents a hitherto unknown genus. Based on both phenotypic and phylogenetic considerations, it is proposed that the unknown bacterium from pig manure be classified in a new genus and species, as Atopostipes suicloacale gen. nov., sp. nov. The type strain of Atopostipes suicloacale is PPC79(T)=NRRL 23919(T)=DSM 15692(T). PMID- 16701519 TI - Subdoligranulum variabile gen. nov., sp. nov. from human feces. AB - During studies on the microflora of human feces we have isolated a strictly anaerobic, non-spore-forming, Gram-negative staining organism which exhibits a somewhat variable coccus-shaped morphology. Comparative 16S ribosomal RNA gene sequencing studies show the unidentified organism is phylogenetically a member of the Clostridium leptum supra-generic rRNA cluster and displays a close affinity to some rDNA clones derived from human and pig feces. The nearest named relatives of the unidentified isolate corresponded to Faecalibacterium prausnitzii (formerly Fusobacterium prausnitzii) displaying a 16S rRNA sequence divergence of approximately 9%, with Anaerofilum agile and A. pentosovorans the next closest relatives of the unidentified bacterium (sequence divergence approximately 10%). Based on phenotypic and phylogenetic considerations, it is proposed that the unusual coccoid-shaped organism be classified as a new genus and species, Subdoligranulum variabile. The type strain of S. variabile is BI 114(T) (=CCUG 47106(T)=DSM 15176(T)). PMID- 16701520 TI - In vitro activity of ramoplanin and comparator drugs against anaerobic intestinal bacteria from the perspective of potential utility in pathology involving bowel flora. AB - Susceptibility of intestinal bacteria to various antimicrobial agents in vitro, together with levels of those agents achieved in the gut, provides information on the likely impact of the agents on the intestinal flora. Orally administered drugs that are poorly absorbed may be useful for treatment of intestinal infections and for certain other situations in which intestinal bacteria may play a role. The antimicrobial activity of ramoplanin (MDL 62,198) against 928 strains of intestinal anaerobic bacteria was determined using the NCCLS-approved Wadsworth brucella laked-blood agar dilution method. The activity of ramoplanin was compared with that of ampicillin, bacitracin, metronidazole, trimethoprim/sulfamethoxazole (TMP/SMX), and vancomycin. The organisms tested included Bacteroides fragilis group (n=89), other Bacteroides species (n=16), other anaerobic Gram-negative rods (n=56) anaerobic cocci (n=114), Clostridium species (n=426), and non-sporeforming anaerobic Gram-positive rods (n=227). The overall MIC(90)s of ramoplanin, ampicillin, bacitracin, metronidazole, and vancomycin were 256, 32, 128, 16, and 128 mcg/ml, respectively. Ramoplanin was almost always highly active vs. Gram-positive organisms and relatively poor in activity against Gram-negative organisms, particularly Bacteroides, Bilophila, Prevotella, and Veillonella. Vancomycin was quite similar to ramoplanin in its activity. Ampicillin was relatively poor in activity vs. organisms that often produce beta-lactamase, including most of the Gram-negative rods as well as Clostridium bolteae and C. clostridioforme. Bacitracin was relatively poor in activity against most anaerobic Gram-negative rods, but better vs. most Gram positive organisms. Metronidazole was very active against all groups other than bifidobacteria and some strains of other types of non-sporeforming Gram-positive bacilli. TMP/SMX was very poorly active, with an MIC(90) of >2048 mcg/ml. PMID- 16701521 TI - Application of group specific amplified rDNA restriction analysis to characterize swine fecal and manure storage pit samples. AB - Group specific amplified ribosomal-DNA restriction analysis was evaluated as a method to rapidly assess microbial community structure in swine fecal and manure storage pit samples. PCR primer sequences were evaluated for their specificity to ribosomal DNA from selected bacterial groups by optimizing annealing temperatures and determining specificity using a set of primer target and non-target organisms. A number of primer sets were identified targeting the following groups: Bacteroides-Prevotella, clostridial clusters I and II, clostridial clusters IX and XI, clostridial clusters XIVa and XIVb, Lactobacillus, Desulfovibrionaceae and Streptococcus-Lactococcus, as well as an universal primer set to represent total populations. Each bacterial group was digested with at least three restriction enzymes. We applied the group specific amplified ribosomal-DNA restriction analysis to swine fecal and manure storage pit samples obtained on two separate occasions. Fecal and manure storage pit samples obtained on the same day were more similar to each other than to any other samples. Results were consistent with 16S ribosomal DNA sequencing data from bacterial isolates and clones obtained from swine feces and manure storage pit. The group specific amplified ribosomal-DNA restriction analysis technique was able to rapid detect gross bacterial community differences among swine fecal and manure storage pit samples and determine groups of interest for more detailed examination. PMID- 16701522 TI - Genetic variation in Brachyspira: chromosomal rearrangements and sequence drift distinguish B. pilosicoli from B. hyodysenteriae. AB - Brachyspira pilosicoli and B. hyodysenteriae are anaerobic pathogenic intestinal spirochetes differing in host range and disease manifestations. Little is known about the size, organization, or genetic content of the B. pilosicoli genome and only limited information is available regarding the genetic organization in B. hyodysenteriae. Both B. hyodysenteriae and B. pilosicoli exist as recombinant populations, and this may be due, in part, to an unusual phage-like gene transfer agent, VSH-1. To compare genetic organization in these two species, the number of mapped loci on an existing physical and genetic map of B. hyodysenteriae B78(T) was expanded, and a combined physical and genetic map of B. pilosicoli P43/6/78(T) was constructed. The B. pilosicoli genome size was about 2.5 Mb, nearly 750 kb smaller than the B. hyodysenteriae genome. Several chromosomal rearrangements have contributed to differences in the size, organization, and content of the two bacterial genomes, and such differences may influence the ability of these species to infect different hosts and cause disease. To evaluate these differences further, comparisons were focused on genes thought to contribute to host-parasite interactions. Four genetic loci (bit, fruBC, vspA, and vspH) were found in B. hyodysenteriae, but not in B. pilosicoli, while two genetic loci (clpX and mglB) were found in B. pilosicoli, but not in B. hyodysenteriae. Contrary to a previous study, an intact copy of the hlyA gene, encoding the B. hyodysenteriae beta-hemolysin gene was detected in B. pilosicoli. Although the hlyA genes of these two species were nearly identical, sequence variation was detected in the intergenic region upstream of hlyA that may alter transcription and translation efficiency of this gene in B. pilosicoli. In addition, divergence in genes flanking hlyA may affect the chemical composition of lipid attached to the mature B. pilosicoli HlyA protein resulting in reduced hemolytic activity. PMID- 16701523 TI - Clostridium perfringens type E enteritis in calves: two cases and a brief review of the literature. AB - Toxigenic types of Clostridium perfringens are important causes of enteric disease in domestic animals, although type E is putatively rare, appearing as an uncommon cause of enterotoxemia of lambs, calves, and rabbits. We report here two geographically distinct cases of type E enterotoxemia in calves, and diagnostic findings which suggest that type E may play a significant role in enteritis of neonatal calves. The cases had many similarities, including a history of diarrhea and sudden death, abomasitis, and hemorrhagic enteritis. In both cases, anaerobic cultures of abomasum yielded heavy growth of C. perfringens genotype E. Four percent of > 1000 strains of C. perfringens from cases of enteritis in domestic animals were type E, and all (n=45) were from neonatal calves with hemorrhagic enteritis. Furthermore, type E isolates represented nearly 50% of all isolates submitted from similar clinical cases in calves. Commercial toxoids available in North America have no label claims for efficacy against type E infections. Consideration should be given to type E-associated enteritis when planning for the health care of calves. PMID- 16701524 TI - Cloning and molecular characterization of the beta toxin (phospholipase C) gene of Clostridium haemolyticum. AB - The phospholipase C (PLPC) gene from Clostridium haemolyticum was amplified using the polymerase chain reaction. Primers were selected from a consensus sequence of closely related clostridial PLPC genes and used to amplify an 871-base pair internal segment of the gene. The internal sequence was used to design nested primers that, together with adapter-specific primers, were used to amplify upstream and downstream sequences. The sequences of upstream and downstream segments were aligned with the internal segment to obtain the entire gene sequence. Primers were selected from the aligned sequence, and the entire gene was amplified, and the PCR product was inserted by ligatation into the pCR 2.1 plasmid. An open reading frame that encodes a 399-amino acid protein, containing a 27-amino acid signal sequence, was identified (GenBank Accession Number AF525415). The molecular weight of the active protein was 42869 Da. A 16-amino acid N-terminal sequence, determined by Edman degradation, exactly matched the putative amino acid sequence of the gene product. Together, N-terminal peptide sequencing and tryptic digestion followed by MALDI-ToF mass spectroscopy verified 48% of the amino acid sequences of the active beta toxin. Comparison of the nucleotide and amino acid sequences with Gene-bank databases demonstrated that the beta toxin of C. haemolyticum exhibits high homology with other bacterial PLPCs. The N-terminal portion of the beta toxin contains zinc-binding residues common to clostridial and other bacterial PLPCs, and it shows 34% homology to the N-terminal domain of bovine arachidonate 5-lipoxygenase. The C-terminal domain of the beta toxin protein shows considerable homology with the C-terminal domains of C. novyi type A PLPC, C. perfringens alpha toxin, C. bifermentens PLPC, although the percent identity between the N-terminal regions is much higher overall than that in the C-terminal domain. PMID- 16701525 TI - Antimicrobial susceptibilities of Bacteroides fragilis and Bacteroides thetaiotaomicron strains isolated from clinical specimens and human intestinal microbiota. AB - Species of Bacteroides fragilis group bacteria are the most prevalent pathogens and have the highest resistance rates to antimicrobial agents among anaerobic bacteria. Infections due to these micro-organisms often originate from patient's own intestinal microbiota. The objective of the study was to determine and compare the susceptibility profiles of clinical and intestinal B. fragilis and B. thetaiotaomicron strains against certain antimicrobials. Isolates were identified by conventional methods and API-20 A. Susceptibility tests were performed according to recommendations of NCCLS (M 11-A4) agar dilution methods. Beta lactamase production was determined with nitrocefin discs. Forty-five clinical isolates (33 B. fragilis and 12 B. thetaiotaomicron) were from following sites: blood (n:8), intra-abdominal abscess (n:7), soft tissue (n:26), and miscellaneous foci of infection (n:4). Fifty B. fragilis and 60 B. thetaiotaomicron isolates from intestinal microbiota of individuals with no history of antimicrobial treatment within last 30 days were also examined. Beta-lactamase production was detected in 93% of clinical and 99% of intestinal isolates. The organisms including intestinal isolates were uniformly susceptible to metronidazole. The MIC90s of other antibiotics and resistance rates of all clinical isolates to those antibiotics were as follows: 256 microg/mL (93%) for ampicillin, 128 microg/mL (13%) for piperacillin, 64 microg/mL (11%) for cefoxitin, 1 microg/mL (2%) for amoxicillin-clavulanate, 0.5 microg/mL (2%) for imipenem, >256 microg/mL (36%) for clindamycin, 8 microg/mL (2%) for chloramphenicol. Intestinal isolates demonstrated similar resistance rates and MIC90s. Metronidazole, imipenem, amoxicillin-clavulanate seem to be effective drugs against these bacteria in Turkey. PMID- 16701526 TI - Anaerobic microbiology in 198 cases of pleural empyema: a Bulgarian study. AB - The aim of the study was to evaluate the incidence of anaerobic bacteria in 198 patients with pleural empyema and the susceptibility of isolates to eight antibacterial agents. Isolates were identified by the Crystal anaerobes identification system, API System rapid ID 32 A and/or routine methods. Susceptibility was tested by Sceptor MIC system for anaerobic bacteria and limited agar dilution method. Anaerobic bacteria were found in 74.2% of the patients and included 247 strains within 21 genera. The predominant anaerobes were Gram-positive anaerobic cocci (52 isolates), Fusobacterium (51), microaerophilic streptococci (24), Prevotella (19) and Bacteroides species (11). Common species/groups were Fusobacterium nucleatum (in 27.2% of specimens yielding anaerobes), Micromonas micros (8.2%), Finegoldia magna (7.5%), Bacteroides fragilis group (6.8%), Peptostreptococcus anaerobius (6.1%) and F. necrophorum (5.4%). No resistance to chloramphenicol and ampicillin/sulbactam was detected. The susceptibility rates of Gram-negative anaerobic isolates to penicillin, cefoxitin, clindamycin, clarithromycin, metronidazole and tetracycline were 63.8%, 90.2%, 87.8%, 58.6%, 98.8% and 71%, and those of Gram positive anaerobes were 79.2%, 100%, 84.3%, 68.4%, 41.9% and 75%, respectively. The wide diversity of isolated anaerobic genera and species and the susceptibility patterns of the isolates emphasize the role of the anaerobic microbiology in cases of pleural empyema. PMID- 16701527 TI - Myristic acid supports the immediate inhibitory effect of lauric acid on ruminal methanogens and methane release. AB - Two in vitro experiments were carried out with the Hohenheim gas test (HGT) apparatus in order to investigate dose-dependent effects and interactions of non esterified lauric acid (C(12)) and myristic acid (C(14)) given either individually or in mixture on ruminal methanogens and methanogenesis. Special emphasis was also put on the relationship between effects on methane formation and methanogenic counts. The in vitro incubations were conducted in 10mL ruminal fluid and 20mL buffer solution and lasted for 24h. In the first experiment, 14 levels of C(12), C(14) and stearic acid (C(18); control) were supplied each in increasing steps of 2.5mg covering the range from 0 to 32.5mg. In the second experiment, dosages ranging from 2.5 to 30mg C(12) were supplemented in steps of 2.5mg either without or with 10, 20 or 30mg of C(14). Counts of total Archaea and individual methanogenic orders were determined by the fluorescence in situ hybridization technique using 16S rRNA oligonucleotide probes. In experiment 1, a methane-suppressing effect of more than 80% was achieved with a supply of 30mg C(12), whereas C(14) and C(18) had no effect. Incubation liquid counts of total Archaea and individual methanogenic orders (Methanococcales, Methanosarcinales, Methanomicrobiales and Methanobacteriales) exponentially decreased as a response to C(12) and C(14) to about the same degree (up to 90%) and, to a lesser extent, by C(18). The proportions of the orders of total methanogenic population were not altered by any of the fatty acids. In experiment 2, an additional supply of 10 or 20mg of C(14) supported the suppression of methanogenesis and methanogens by C(12) synergistically. Supplementing 30mg instead of 20mg of C(14) did not further increase the efficacy of C(12) in suppressing methane formation and methanogens. The study illustrated the advantage of using mixtures of C(12) and C(14) in ruminant nutrition to suppress methane emission since mixtures will reduce the amounts of the less palatable C(12) required in feed. PMID- 16701528 TI - 16S ribosomal DNA-directed PCR primers for ruminal methanogens and identification of methanogens colonising young lambs. AB - The population densities and identities of methanogens colonising new-born lambs in a grazing flock were determined from rumen samples collected at regular intervals after birth. Methanogen colonisation was found at the first sampling (1 3 days after birth) and population densities reached around 10(4) methanogens per gram at 1 week of age. Population densities increased in an exponential manner to a maximum of 10(8)-10(9) per gram at 3 weeks of age. To identify methanogens, PCR primers specific for each of the Archaea; a grouping of the orders Methanomicrobiales, Methanosarcinales and Methanococcales; the order Methanobacteriales; the order Methanococcales; the order Methanosarcinales; the genus Methanobacterium; and the genus Methanobrevibacter were designed. Primer pair specificities were confirmed in tests with target and non-target micro organisms. PCR analysis of DNA extracts revealed that all the detectable ruminal methanogens belonged to the order Methanobacteriales, with no methanogens belonging to the Methanomicrobiales, the Methanosarcinales, or the Methanococcales being detected. In 3 lambs, the initial colonising methanogens were Methanobrevibacter spp. and in 2 lambs were a mixture of Methanobrevibacter and Methanobacterium spp. In the latter case, the initial colonising Methanobacterium spp. subsequently disappeared and were not detectable 12-19 days after birth. Seven weeks after birth, lambs contained only Methanobrevibacter spp. This study, the first to provide information on the identities of methanogens colonising pre-ruminants, suggests that the predominant methanogens found in the mature rumen establish very soon after birth and well before a functioning rumen develops. PMID- 16701529 TI - Influence of nitrate and COD on phosphorus, nitrogen and dinitrotoluene (DNT) removals under batch anaerobic and anoxic conditions. AB - In this study, the effects of COD to NO(3)-N ratio in the feed on PO(4)-P removal was investigated. Maximum PO(4)-P uptake was obtained in the anoxic reactor when the COD to NO(3)-N ratios were between 2 and 3.75. With the influent COD of 800 1500 mg COD/L a total of the maximum removable PO(4)-P was 56 mg PO(4)-P/L through 20 days of anaerobic/anoxic incubation, indicating 98% P removal in the anoxic reactor. Similarly, for the COD to NO(3)-N ratios varying between 2 and 3.75 maximum denitrification was observed. Through anoxic operation the poly-P bacteria are capable of removing NO(3)-N using VFA, COD as carbon source and NO(3)-N as the electron acceptor after methanogenesis has been completed. High NO(3)-N concentrations stopped significantly the P uptake. A total of 97-99% dinitrotoluene removal efficiencies in the reactors containing COD to NO(3)-N ratio of 2 and 3.75 after 20 days of incubation period. For maximum NO(3)-N and PO(4)-P removals optimal COD to NO(3)-N ratios, COD and NO(3)-N concentrations were 2-3.75, 2000-4000 mg COD/L and, 800-1500 mg NO(3)-N/L, respectively. PMID- 16701530 TI - Inhibition of Clostridium perfringens sporulation by Bacteroides fragilis and short-chain fatty acids. AB - The effect a common fecal organism, Bacteroides fragilis, has on the sporulation of Clostridium perfringens, an organism linked to some cases of antibiotic associated diarrhea, was examined. Established B. fragilis cultures significantly decreased the number of heat resistant spores formed by C. perfringens ATCC 12915 and increased the number of vegetative cells. To determine if short-chain fatty acids (SCFA), fermentation products of B. fragilis, inhibited sporulation, the SCFA were added to sporulation broth. Sporulation decreased in the presence of acetate, isobutyrate, isovalerate, and succinate. Vegetative cell number for C. perfringens decreased in the cultures with isobutyrate. Propionate did not affect sporulation or vegetative cell number. The data support the hypothesis that the decrease in short-chain fatty acid concentration following antibiotic therapy predisposes patients to diarrheas caused by C. perfringens. PMID- 16701531 TI - Allobaculum stercoricanis gen. nov., sp. nov., isolated from canine feces. AB - Morphological, biochemical, and molecular genetic studies were performed on an unknown anaerobic, catalase-negative, non-spore-forming, rod-shaped bacterium isolated from dog feces. The unknown bacterium was tentatively identified as a Eubacterium species, based on cellular morphological and biochemical tests. 16S rRNA gene sequencing studies, however, revealed that it was phylogenetically distant from Eubacterium limosum, the type species of the genus Eubacterium. Phylogenetically, the unknown species forms a hitherto unknown sub-line proximal to the base of a cluster of organisms (designated rRNA cluster XVI), which includes Clostridium innocuum, Streptococcus pleomorphus, and some Eubacterium species. Based on both phenotypic and phylogenetic criteria, it is proposed that the unknown bacterium be classified as a new genus and species, Allobaculum stercoricanis. Using a specific rRNA-targeted probe designed to identify Allobaculum stercoricanis, in situ hybridisation showed this novel species represents a significant organism in canine feces comprising between 0.1% and 3.7% of total cells stained with DAPI (21 dog fecal samples). The type strain of Allobaculum stercoricanis is DSM 13633(T)=CCUG 45212(T). PMID- 16701532 TI - Changes in taxonomy, anaerobes associated with humans, 2001-2004. PMID- 16701533 TI - Phylogenetic analysis of archaea in three fractions of cow rumen based on the 16S rDNA sequence. AB - Phylogenetic analysis of archaea in the rumen ecosystem was analysed by PCR of 16S rDNA from the bovine rumen using archaea-specific primers. The libraries were constructed from rumen fluid (AF), rumen solid (AS), and rumen epithelium (AE) from a rumen-fistulated Korean cow (Hanwoo). The 45 AF clones could be divided into three groups and the largest group was affiliated with the Methanomicrobiaceae family (96% of clones). The AF clones contained a high proportion of unidentifiable clones (67%). The 39 AE clones could be divided into two groups and the largest group was also affiliated with the Methanomicrobiaceae family (95% of clones). The AE clones contained a low proportion of unidentifiable clones (5%). The 20 AS clones could be divided into two groups that were affiliated with either the Methanobacteriaceae family (55%) or the Methanomicrobiaceae family (45%). The AS clones contained a moderate proportion of unidentifiable clones (40%). The predominant family of whole rumen archaea was found to belong to the Methanomicrobiaceae (85%). Methanomicrobiaceae were predominant in the rumen epithelium and the rumen fluid while Methanobacteriaceae were predominant in the rumen solid. One clone from the rumen fluid and two clones from the rumen epithelium contained rDNA sequences of Non-Thermophilic Crenarchaeota (NTC) and Thermophilic-Crenarchaeota (TC), respectively, which have not previously been described from the rumen. PMID- 16701535 TI - In vitro growth and acid production by mutans streptococci on traditional African foods. AB - The growth rate and production of acids by mutans streptococci (MS) are influenced by their ability to ferment different dietary carbohydrates. This suggests that the nutrient environment in the oral cavity affects bacterial virulence. The aim of this study was to investigate the effect of maize, samp and brown bread on the growth and acidogenicity of this species. Six laboratory references and five clinical strains isolated from the dental plaque of South African black and 'colored' (historical race classification) children were studied in batch culture on maize, samp (coarsely ground maize), brown bread and compared against a 3% sucrose control. The doubling time of bacterial strains was prolonged in maize (1.9-17.5 h) and samp (2.4-18.4 h), and the number of cell divisions was low. Staple foods accounted for 25% (F=5.98; P=0.0007) and MS strains 30.78% (F=2.84; P=0.009) of the total variance. The fermentation of samp and maize showed the least drop in pH of the culture medium, ranging between 0.54 and 1.06 and 0.69 and 2.28 pH units respectively, with variation between strains most significant in maize (F=33.62; P<0.0001). The total mean concentration of acids produced was highest in bread (25.13 mM/mL) and samp (17.00 mM/mL) which was comparable to Brain Heart Infusion broth (16.49 mM/mL) and a basal synthetic medium (17.96 mM/mL) containing 3% sucrose, but the yield of lactate, acetate and formate was low during the fermentation of samp (0.50 mM/mL), BHI+3% sucrose (4.12 mM/mL) and brown bread (0.06 mM/mL) respectively. Results indicated that maize and samp do not optimally support the growth or acid production by MS, and the varying response of test strains demonstrates the strain variability of this species to different carbohydrate sources in the diet. PMID- 16701534 TI - Regulation of neurotoxin complex expression in Clostridium botulinum strains 62A, Hall A-hyper, and NCTC 2916. AB - The kinetics of botulinum toxin gene expression have been investigated in Clostridium botulinum type A strains 62A, Hall A-hyper, and type A(B) strain NCTC 2916 during the growth cycle. The analyses were performed in TPGY and type A Toxin Production Media (TPM). The mRNA transcript levels encoding the proteins of the neurotoxin complex were determined using Northern analyses. Neurotoxin concentrations in culture supernatants and lysed cell pellets were assayed using ELISA, Western blots, and mouse bioassay. Proteolytic activation of botulinum neurotoxin during the growth cycle was evaluated by Western blots. For all three strains, mRNA transcripts for the toxin complex genes were initially detected in early log phase, reached peak levels in early stationary phase, and rapidly decreased in mid-to-late stationary phase and during lysis. Toxin expression varied depending on the strain and growth medium. Toxin production was highest in strain Hall A-hyper, followed by NCTC 2916 and 62A. For C. botulinum strain Hall A-hyper, cell lysis and toxin release into the supernatant occurred rapidly for cells grown in TPM, while cells grown in TPGY remained in stationary phase with minimal lysis and toxin release through 96 h of growth. In contrast, strains 62A and NCTC 2916 lysed more extensively than Hall A-hyper in TPGY. TPM supported higher toxin production and activation than TPGY in strains 62A and Hall A-hyper. These data support that the genes of the botulinum neurotoxin complex are temporally expressed during late-log and early stationary phase and that toxin complex formation depends on the strain and growth medium. Botulinum toxin synthesis and activation appears to be a complex process that is highly regulated by nutritional and environmental conditions. Further research is needed to elucidate the sensing mechanisms and genetic regulatory factors controlling these processes. PMID- 16701536 TI - PCR-based diagnostics for anaerobic infections. AB - Conventional methods to identify anaerobic bacteria have often relied on unique clinical findings, isolation of organisms, and laboratory identification by morphology and biochemical tests (phenotypic tests). Although these methods are still fundamental, there is an increasing move toward molecular diagnostics of anaerobes. In this review, some of the molecular approaches to anaerobic diagnostics based on the polymerase chain reaction (PCR) are discussed. This includes several technological advances in PCR-based methods for the detection, identification, and quantitation of anaerobes including real-time PCR which has been successfully used to provide rapid, quantitative data on anaerobic species on clinical samples. Since its introduction in the mid-1980s, PCR has provided many molecular diagnostic tools, some of which are discussed within this review. With the advances in micro-array technology and real-time PCR methods, the future is bright for the development of accurate, quantitative diagnostic tools that can provide information not only on individual anaerobic species but also on whole communities. PMID- 16701537 TI - In vitro activity of oritavancin (LY333328), vancomycin, clindamycin, and metronidazole against Clostridium perfringens, Propionibacterium acnes, and anaerobic Gram-positive cocci. AB - Using an agar dilution method, we determined the in vitro activity of oritavancin, vancomycin, clindamycin and metronidazole against 114 unique clinical isolates of Gram-positive anaerobes. MIC(90)s (microg/mL) for oritavancin were as follows: Clostridium perfringens 1.0, Propionibacterium acnes 0.25, Peptostreptococcus anaerobius 0.25, Peptoniphilus asaccharolyticus 0.5, Finegoldia magna 0.25, Micromonas. micros 0.25, and Anaerococcus prevotii 0.25. On a weight basis, oritavancin is slightly more active than vancomycin against the strains tested. The oritavancin MICs are comparable to those previously reported against staphylococci and enterococci. Oritavancin shows excellent potential for treatment of infections containing Gram-positive anaerobes such as these. PMID- 16701538 TI - Fatal septicemia due to Clostridium hathewayi and Campylobacter hominis. AB - Clostridium hathewayi and Campylobacter hominis have not been previously reported in infection. We report a fatal case of septicemia, massive intravascular hemolysis, shock, and disseminated intravascular coagulation; both of these organisms were recovered on blood culture, although it seems likely that the C. hathewayi was responsible for the clinical picture and that the C. hominis was an incidental finding. PMID- 16701539 TI - Enzymatic/biochemical analysis of Actinomyces with commercial test kits with an emphasis on newly described species. AB - In clinical microbiology laboratories, the identification of Actinomyces-like bacteria can be very laborious and problematic. In the present study, we focused on reactivity patterns of 4 commercial test kits, RapID ANA II, RapID 32A, RapID CB Plus, and BBL Crystal ANR ID, that could be used for rapid preliminary identification of Actinomyces isolates belonging to newly described Actinomyces and closely related species. Out of the 54 strains tested, 25 strains (46%) were correctly identified to the genus/group level by BBL Crystal ANR ID system, 16 strains (30%) by RapID 32 A, 11 strains (20%) by RapID CB Plus, and 7 strains (13%) by RapID ANA II. The main problems with these kits were due to occasional weak enzymatic and sugar fermentation reactions. In conclusion, chromogenic substrate sensitivity and specificity need to be enhanced in order to improve the reliability of the test results of these kits, and the present database updated in order to more precisely identify newly described Actinomyces and closely related species. PMID- 16701540 TI - Diarrhea caused by enterotoxigenic Bacteroides fragilis in children less than 5 years of age in Hanoi, Vietnam. AB - Enterotoxigenic Bacteroides fragilis (ETBF) are considered as an emerging enteropathogen causing diarrhea in children. Eight hundred and thirty-six (836) children less than 5 years of age including 587 children with diarrhea and 249 age-matched controls were involved in the study. Within the group of children with diarrhea, 7.3% (43/587) ETBF was detected by immunoseparation in combination with polymerase chain reaction. The corresponding figure for the controls was 2.4% (6/249) (P<0.01). Within the diarrhea group, the prevalence was significantly higher in children older than 1 year of age. Three subtypes of ETBF isolates have been identified with the prevalence of 67.4%, 18.6%, and 16% for bft-1, bft-2, and a new bft, respectively. In the controls, two of the subtypes were identified, 5 bft-1 and 1 bft-2. More than half (55.8%) of the samples harboring ETBF also had other identified pathogens. The clinical symptoms of the single ETBF infection were not different from those of co-infections. This is the first study of the role of ETBF in children's diarrhea in Vietnam and it is concluded that this pathogen is an important causative agent of diarrhea in children in Hanoi, Vietnam. PMID- 16701541 TI - Cloning and sequencing of the genes for A and B subunits of the V-type Na(+) ATPase of a facultatively anaerobic alkaliphile. AB - The structural genes for A and B subunits of the V-type Na(+)-ATPase from a facultatively anaerobic alkaliphile (Amphibacillus sp.), strain M-12, were cloned and sequenced. Transformation of Escherichia coli with the genes overexpressed two proteins, which crossreacted with an antiserum against A and B subunits of the V-type Na(+)-ATPase from Enterococcus hirae. The deduced amino acid sequence (594 amino acids; Mr, 66,144) of A subunit of the M-12 enzyme exhibited 73%, 51%, 49% and 53% identities with those of V-type ATPases from E. hirae, Thermus thermophilus, Neurospora crassa and Drosophila melanogaster, respectively. The amino acid sequence (458 amino acids; Mr, 51,308) of B subunit of the M-12 enzyme was 74%, 53%, 52% and 54% identical with those of the ATPases from E. hirae, T. thermophilus, N. crassa and D. melanogaster, respectively. The fact indicates that the amino acid sequences of A and B subunits of the M-12 enzyme exhibit significantly higher homologies with those of the E. hirae Na(+)-ATPase as compared with those of the H(+)-ATPases from T. thermophilus, N. crassa and D. melanogaster. PMID- 16701542 TI - The leukotoxin operon of Fusobacterium necrophorum is not present in other species of Fusobacterium. AB - The complete nucleotide sequence of the leukotoxin operon of Fusobacterium necrophorum has been determined. The operon consists of three genes (lktBAC) of which the leukotoxin structural gene is the middle determinant. Southern and western blot analyses and flow cytometry analysis for biological activity of the culture supernatants were carried out to determine if the leukotoxin is present in other species of the genus Fusobacterium. Only the two subspecies of F. necrophorum were found to possess the leukotoxin locus and produce the toxin. The human periodontal pathogen, F. nucleatum does not produce detectable leukotoxin. The F. necrophorum leukotoxin was found to be active against human neutrophils. PMID- 16701543 TI - Anaerobes in the upper respiratory tract in infancy. AB - Development of the indigenous microbiota begins on the surfaces of the human body after birth when infants are exposed to continuous person-to-person and environmental contacts with microbes. Anaerobes constitute a significant part of indigenous bacterial communities at different body sites. Pioneering anaerobic commensals are able to colonize and survive in the oral cavity during the first months of life. After teeth emerge, more attachment sites and potential niches are available for anaerobic bacterial colonization. Specific partner relationships influence the composition and stability of forming multigeneric communities, biofilms, where Fusobacterium nucleatum is of specific interest. In infancy, the oral colonization seems to be rather stable at species level, though not at clonal level. The colonization pattern in the nasopharynx is different from that in the oral cavity; anaerobes are absent from healthy nasopharynges but transiently colonize this anatomical site during infection. The most plausible origin for nasopharyngeal anaerobes is the oral cavity and, conceivably, saliva is the most likely transmission vehicle. Whether anaerobic bacteria colonize the nasopharynx just because of ecological changes favoring their growth or whether they could play an active role in the pathogenesis of respiratory infections is not known. PMID- 16701544 TI - Genetic basis of resistance in Propionibacterium acnes strains isolated from diverse types of infection in different European countries. AB - The purpose of the study was to characterize the resistance mechanism of 36 clindamycin (CL) and erythromycin (EM) resistant Propionibacterium acnes strains and 27 tetracycline (TET) resistant P. acnes isolates, collected from nine European countries, both from acne patients and from patients with different infections. PCR and sequencing of the genes encoding domain V of 23S rRNA for CL and EM resistant strains and 16S rRNA for TET resistant strains were performed. Pulsed-field gel electrophoresis was used as a typing method to establish the relationship between resistance genotypes and pulsed-field types. Several unique resistant genotypes were found to be distributed throughout Europe. P. acnes CL and EM resistant strains carrying one of the mutations within the 23S rRNA were predominantly isolated from Swedish acne patients (64%) compared to other infections (43%), OR=2.33 [CI=1.16-4.69]. Of 36 P. acnes isolates tested, none was found to carry the erm(X) resistance gene. Forty-four percent of TET resistant strains were found to carry a G-C transition in the 16S rRNA of the small ribosomal subunit and all these strains were isolated from Swedish acne patients. MIC of TET among all strains carrying this G-C mutation (n=12) was 32 mg/L and the MIC range for the strains where no mutation was detected ranged from 2 to 8 mg/L. The MIC values of TET were unaffected by the presence of reserpine, a well-known inhibitor of efflux pumps. Those TET resistant strains harbouring the mutation at 16S rRNA were clustered in one pulsotype. For TET resistant strains where no mutation was found, greater variability was noticed. No correlation was noticed between different resistance genotypes of CL and EM resistant strains and pulsed-field types. PMID- 16701545 TI - Prebiotic effects of oligosaccharides on selected vaginal lactobacilli and pathogenic microorganisms. AB - The purpose of this study was to select endogenous human vaginal lactobacilli strains on the basis of the main probiotic properties observed in the vaginal environment in order to use them for the evaluation of the potential prebiotic properties of oligosaccharides. From vaginal samples of 50 women with a normal flora, 17 lactobacilli strains were first isolated because of their high level of hydrogen peroxide production. Then six strains were selected mainly for their ability (i) to adhere to vaginal cells, (ii) to produce compounds in sufficient amount, such as lactic acid, having an inhibitory action on pathogens, and less importantly, (iii) to demonstrate arginine deiminase activity. These six strains were found to belong to three distinct species: Lactobacillus crispatus, L. jensenii and L. vaginalis. One strain of each species was chosen as a potential vaginal probiotic strain with regard to our criteria. These three strains were then used to evaluate the prebiotic properties of different oligosaccharide series: two fructooligosaccharide series (FOS Actilight and FOS Raftilose) and two glucooligosaccharide series varying by their osidic linkages (alpha-1,6/alpha 1,4 GOS and alpha-1,2/alpha-1,6/alpha-1,4 GOS). The test was based on the ability of the oligosaccharides to promote the growth of the three beneficial strains selected but not of pathogenic microorganisms often encountered in urogenital infections such as Candida albicans, Escherichia coli and Gardnerella vaginalis. Oligosaccharide hydrolysis was followed by HPLC analysis. This revealed that two oligosaccharide series (FOS Actilight DP3 and all alpha-1,6/alpha-1,4 GOS DP > or = 4) were used only by the lactobacilli strains, the pathogenic microorganisms being unable to metabolise them. The selected lactobacilli and oligosaccharides are good candidates for incorporation in a formula to prevent vaginal infections. PMID- 16701546 TI - Optimal protease production condition for Prevotella ruminicola 23 and characterization of its extracellular crude protease. AB - In this study, Prevotella ruminicola 23 (ATCC 19189), a ruminal proteolytic bacterium, was used as protease producer to examine the optimal condition for protease production. The best carbon and nitrogen sources for the maximum growth were glucose with peptone. Both sucrose and glucose could stimulate high protease production. Casein and peptone are better nitrogen sources for protease production than other choice in this study. The best enzyme production condition was 18-20 h incubation which was at late log phase in the broth of 5% glucose or sucrose as carbon source with 0.1% ammonium chloride and 0.2% peptone as nitrogen sources. Most of the protease activity was secreted into broth (65%) and on cell surface (18%). The optimal temperature and pH for protease reaction were 40 degrees C and pH 6.8, respectively. After incubation for 6h, the crude extract maintained 50% of original protease activity at 30 and 50 degrees C, and protease activity was stable between pH 6 and 8. The protease inhibitor test showed that serine, aspartic acid and metallo-protease inhibitors could cause inhibition of proteolysis. Protein feedstuff degradation experiments suggested that protease in crude extract had higher degradation ability on fish meal, whey, and feather meal (2.39, 2.60 and 1.76 micromol aminoacid/mg enzyme/h) in comparison to soybean meal and blood meal (1.11 and 1.09 micromol aminoacid/mg enzyme/h). The protease in the crude extract should have application potential in term of improving utilization of fish meal and feather meal for monogastric animals. PMID- 16701547 TI - Relative fibrolytic activities of anaerobic rumen fungi on untreated and sodium hydroxide treated barley straw in in vitro culture. AB - The fibrolytic activities of rumen fungi were studied in terms of dry matter loss, plant cell wall degradation and enzyme (cellulase and xylanase) activities, when grown in vitro on either untreated or sodium hydroxide treated stems of barley straw over a 12 day period. Changes in fungal growth, development and overall biomass were followed using chitin assay and scanning electron microscopy. Treatment with sodium hydroxide resulted in a decrease in the NDF content together with the disruption of cuticle and the loosening and separation of the plant cells within the straw fragments. The enzyme activities of the anaerobic fungi have a high positive correlation (R(2)=0.99) with their biomass concentration assessed by chitin assay indicating that chitin is a valuable index for the estimation of the fungal biomass in vitro. The anaerobic fungi produced very extensive rhizoidal systems in these in vitro cultures. After incubation with rumen fungi, dry matter losses were, respectively, 35% and 38% for the untreated and treated straw samples and the overall fungal biomass, determined by chitin assay, was significantly higher in the treated samples. In vitro degradation of cellulose and hemicellulose was also higher in the treated than that of untreated cultures. Although, comparatively, xylanase activity was higher than that of cellulase, the cellulose fraction of the straw was degraded more than hemicellulose in both treated and untreated straw. PMID- 16701548 TI - Enrichment of fusobacteria from the rumen that can utilize lysine as an energy source for growth. AB - Ruminal lysine degradation is a wasteful process that deprives the animal of an essential amino acid. Mixed ruminal bacteria did not deaminate lysine (50 mM) at a rapid rate, but lysine degrading bacteria could be enriched if Trypticase (5 mg/mL) was also added. Lysine degrading isolates produced acetate, butyrate and ammonia, were non-motile, stained Gram-negative and could also utilize lactate, glucose, maltose or galactose as an energy source for growth. Lactate was converted to acetate and propionate, and 16S rDNA indicated that their closest relatives were Fusobacterium necrophorum. Growing cultures produced ammonia at rates as high as 2400 nmol/mg protein/mL/min. Washed cell suspensions took up (14)C lysine (3 microM) at an initial rate of 6 nmol/mg protein/min, and glucose addition did not affect the transport. Cells washed aerobically had the same transport rate as those handled anaerobically, but only if the transport buffer contained sodium. The affinity constant for sodium was 8 mM, and sodium could not be replaced by lithium. Cells treated with the sodium/proton antiporter, monensin (5 microM), did not take up lysine, but a protonophore that inhibited growth (tetrachlorosalicylanilide, 10 microM) had no effect. An artificial membrane potential created by potassium diffusion did not increase the rate of lysine transport, and an Eadie-Hofstee plot indicated the transport rate was directly proportional to the lysine concentration. Decreasing the pH from 6.7 to 5.5 caused an 85% decrease in the rate of lysine transport. The addition of F. necrophorum JB2 (130 microg protein/mL) to mixed ruminal bacteria increased lysine degradation 10-fold, but only if the pH was 6.7 and monensin was not present. Further work will be needed to see if dietary lysine enriches fusobacteria in vivo. PMID- 16701549 TI - A case of Lemierre's syndrome following Epstein-Barr virus infection. AB - Lemierre's syndrome, or necrobacillosis, is a life-threatening septic thrombophlebitis of the internal jugular vein. The causative organism is Fusobacterium necrophorum. Here we report a case of Lemierre's syndrome in a young male and describe the clinical presentations and treatment. Epstein-Barr virus (EBV) was a suspected predisposing factor in this case. PMID- 16701550 TI - Hyperactivation of midbrain dopaminergic system in schizophrenia could be attributed to the down-regulation of dysbindin. AB - Extraordinal activation of nigrostriatal and mesolimbic dopaminergic systems (midbrain dopaminergic system) is thought to be one of the most important etiologies for schizophrenia, though the reason why unusual hyperactivation of the dopaminergic system occurs in the schizophrenic brain is quite obscure. Dysbindin, one of the most susceptible genes for schizophrenia, has been reported to be reduced in the schizophrenic brain. In situ hybridization analysis showed the mRNA expression of dysbindin in the mouse substantia nigra. Furthermore, suppression of dysbindin expression in PC12 cells resulted in an increase of the expression of SNAP25, which plays an important role in neurotransmitter release, and increased the release of dopamine. On the other hand, up-regulation of dysbindin expression in PC12 cells showed a tendency to decrease the expression of SNAP25. These data suggest that dysbindin might regulate the dopamine release of the dopaminergic system via modulation of the expression of SNAP25. PMID- 16701551 TI - Phosphatidylserine is involved in gene expression from Sindbis virus subgenomic promoter. AB - Sindbis virus replication is mediated by an RNA replicase translated from viral RNA genome. The replicase synthesizes progeny genomic RNA and shorter RNA (subgenomic RNA) carrying viral structural genes in association with cytoplasmic membranes. Here we examined the involvement of a membrane lipid, phosphatidylserine (PS), in Sindbis virus gene expression using Chinese hamster ovary cell mutants. When the mutant cells were transfected with viral replicon RNA, in which the structural genes downstream of the subgenomic promoter were replaced by a reporter gene, reporter expression was inhibited under PS-deficient conditions. In contrast, reporter expression from an SV40 promoter-driven construct was normal under similar conditions. Furthermore, expression of a viral replicase protein from the genomic RNA and accumulation of the subgenomic RNA were not inhibited by PS deficiency. These findings indicate that reduced cellular PS level impairs a posttranscriptional event of Sindbis virus subgenomic promoter-driven gene expression. PMID- 16701553 TI - Characterization of the cheY genes from Leptospira interrogans and their effects on the behavior of Escherichia coli. AB - The motility and chemotaxis system are critical for the virulence of pathogenic leptospire, which enable them to penetrate host tissue barriers during infection. The completed genome sequence of a representative virulent serovar type strain (Lai) of Leptospira interrogans serogroups Icterohaemorrhagiae (L. interrogans strain Lai) suggested that there were multiple copies of putative chemotaxis homologues located at its large chromosome. In order to verify the function of these proteins, the putative cheY genes were cloned into pQE31 vector and then expressed, respectively, in wild-type Escherichia coli strain RP437 and cheY defective strain RP5232. The results showed that all the five cheYs could restore the swarming of RP5232 strain to some extend. Overexpression of CheYs in RP437 showed inhibited swarming of RP437. To investigate the mechanism of chemotaxis signaling in L. interrogans strain Lai, certain aspartates (Asp-53, Asp-61, Asp 70, Asp-62, and Asp-66 for L. interrogans strain Lai CheY1, CheY2, CheY3, CheY4, and CheY5, respectively) were mutated. Expression of these mutated cheYs manifested neither restoration of the swarming ability of RP5232 nor inhibition on swarming ability of RP437. Multiple amino acid sequence alignment predicted ternary structures and the result of mutation experiment suggested that these conserved aspartate residues of L. interrogans were analogous to that in E. coli CheY in function and structure. So, L. interrogans and E. coli may have similar mechanisms of activation of the chemotaxis phosphorelay pathway, but there are differences in their control by signal terminator. PMID- 16701552 TI - Characterization of the protease activity that cleaves the extracellular domain of beta-dystroglycan. AB - Dystroglycan (DG) complex, composed of alphaDG and betaDG, provides a link between the extracellular matrix (ECM) and cortical cytoskeleton. Although the proteolytic processing of betaDG was reported in various physiological and pathological conditions, its exact mechanism remains unknown. In this study, we addressed this issue using the cell culture system of rat schwannoma cell line RT4. We found that the culture medium of RT4 cells was enriched with the protease activity that degrades the fusion protein construct of the extracellular domain of betaDG specifically. This activity was suppressed by the inhibitor of matrix metalloproteinase-2 (MMP-2) and MMP-9, but not by the inhibitors of MMP-1, MMP-3, MMP-8, and MMP-13. Zymography and RT-PCR analysis showed that RT4 cells secreted MMP-2 and MMP-9 into the culture medium. Finally, active MMP-2 and MMP-9 enzymes degraded the fusion protein construct of the extracellular domain of betaDG. These results indicate (1) that RT4 cells secrete the protease activity that degrades the extracellular domain of betaDG specifically and (2) that MMP-2 and MMP-9 may be involved in this process. PMID- 16701554 TI - Impact of L-arginine on hydrogen sulfide/cystathionine-gamma-lyase pathway in rats with high blood flow-induced pulmonary hypertension. AB - The present study was designed to explore the possible effect of L-arginine on endogenous hydrogen sulfide/cystathionine-gamma-lyase (H(2)S/CSE) pathway in the pathogenesis of pulmonary hypertension and pulmonary vascular structural remodeling induced by high pulmonary blood flow. Thirty-two male Sprague-Dawley rats were randomly divided into control group (n=11), shunt group (n=11) and shunt with L-arginine group (n=10). Rats in the shunt and shunt with L-arginine group underwent an abdominal aorta-inferior cava vein shunt operation. After 11 weeks of shunting, the plasma level of H2S and lung tissue H2S production rate in the shunt with L-arginine group were much higher than those in the shunt group (P<0.01). Meanwhile, the expression of CSE mRNA in the lung tissues of rats in the shunt with L-arginine group was increased significantly (P<0.01), and in situ hybridization showed that CSE mRNA expression was obviously up-regulated in the smooth muscle cells (SMCs) of the pulmonary arteries of shunted rats treated with L-arginine when compared with shunted rats without the treatment of L-arginine (P<0.01). In conclusion, H2S/CSE pathway was up-regulated by L-arginine in pulmonary hypertension induced by high blood flow with the attenuation of pulmonary hypertension and pulmonary vascular structural remodeling. PMID- 16701555 TI - Contrasting actions of prolonged mitogen-activated protein kinase activation on cell survival. AB - Activation of the ERK mitogen-activated protein kinase pathway has been implicated in pro-survival and cellular protective mechanisms, so that chronic ERK activation may be a useful therapeutic strategy. Here, we further explored the consequences of prolonged ERK activation following expression of constitutively active form of MEK, MEK-EE, in cardiac myocytes. We confirmed that chronic MEK-EE overexpression halved myocyte death following glucose deprivation, but surprisingly this was not associated with preserved intracellular ATP levels. Whilst activities of a number of antioxidant enzymes were not altered upon MEK-EE expression, paradoxically Cu/Zn superoxide dismutase activity was almost halved upon MEK-EE expression. When we then exposed myocytes to the superoxide generator menadione, we observed significantly higher death of MEK-EE expressing myocytes. Pre-incubation with U0126 inhibited menadione-induced death. Our results are the first to show that MEK-ERK signalling can act to increase or decrease cell survival, the outcome depending on the form of stress stimulus encountered. PMID- 16701556 TI - Analysis of the temperature-sensitive mutation of Escherichia coli pantothenate kinase reveals YbjN as a possible protein stabilizer. AB - Pantothenate kinase (PanK), a key regulatory enzyme in the coenzyme A (CoA) biosynthetic pathway, catalyzes the rate-limiting phosphorylation of pantothenic acid to form phosphopantothenate during CoA biosynthesis. Escherichia coli ts9 strain manifests temperature-sensitive phenotype on LB media due to its mutation in the coaA gene (coaA1). Sequencing analysis revealed that coaA1 arises from a single base pair mutation that results in an amino acid change, L236F. This change, located proximate to the ATP binding site of CoaA, destabilizes both enzymatic activity and structural integrity or stability of the mutant protein in vitro. Spontaneously, revertants of ts9 were occasionally found on LB medium plates. Two groups of revertants were isolated: for those that can grow at 40 degrees C, a reversion of the original amino acid mutation L236F to L236L or other amino acid (such as L236C) occurs; for those that can grow at 37 degrees C but not 40 degrees C, a mutation at another gene or intergenic suppression is strongly indicated. Towards genetic identification of genes that might interact with coaA1, ybjN, which encodes a putative sensory transduction regulator protein, and whose over-expression is capable of ameliorating the temperature sensitive phenotype of the structurally unstable CoaA1 or CoaA[L236F], was isolated. Over-expression of ybjN appears to suppress the temperature-sensitive phenotype of several other temperature-sensitive mutations, including coaA14 (carried by DV51 strain), coaA15 (carried by DV70 strain), and ilu-1, suggesting it not only helps CoaA1, but possibly works as a general stabilizer for some other unstable proteins. PMID- 16701557 TI - Comparative studies of Bombyx mori nucleopolyhedrovirus chitinase and its host ortholog, BmChi-h. AB - Baculovirus-encoded chitinases (V-CHIAs) were first proposed to be acquired from a bacterium via horizontal gene transfer. However, we have recently reported that lepidopteran hosts also encode v-chiA orthologs. Here we describe comparative studies of Bombyx mori nucleopolyhedrovirus (BmNPV) chitinase and its host ortholog, BmChi-h. We constructed recombinant BmNPVs in which native and modified forms of BmChi-h were driven under the polyhedrin promoter and the authentic v chiA was deleted. Western blot analysis indicated that BmCHI-h was rapidly secreted from virus-infected BmN cells whereas BmNPV CHIA was localized within the virus-infected cells; probably because of the presence of a C-terminal endoplasmic reticulum retention motif on BmNPV CHIA. Enzymological studies showed that BmNPV CHIA was able to retain much higher chitinolytic activity under alkaline conditions. For B. mori larvae infected with v-chiA-deleted BmNPV, the terminal liquefaction of dead larvae and the activation of baculovirus-encoded cysteine protease were not observed, and the introduction of BmChi-h did not rescue these defects. Our findings show that BmNPV chiA possesses unique features that are not shared by host orthologs, which may reflect functional specialization of baculovirus chitinases. PMID- 16701559 TI - 5'-end formation of yeast 5.8SL rRNA is an endonucleolytic event. AB - Like most eukaryotes, Saccharomyces cerevisiae cells contain a minor 5.8SL rRNA that, relative to the major 5.8SS species, carries several extra nucleotides at the 5'-end. The two species are produced by alternative pathways that differ in the events removing the 3'-terminal region of Internal Transcribed Spacer 1 from the 27SA2 pre-rRNA. Whereas the pathway leading to 5.8SS rRNA is well established, that producing the 5'-end of 5.8SL (called B1L) is poorly understood. Northern analysis of two different mutants of S. cerevisiae that overproduce 5.8SL rRNA revealed the presence of a fragment corresponding to the 3'-terminal region of Internal Transcribed Spacer 1 (ITS1) directly upstream from site B1L. Immunoprecipitation experiments showed this fragment to be associated with the trans-acting factor Rrp5p required for processing at the early sites A0 A3. Together these data clearly support that the 5'-end of 5.8SL rRNA is an endonucleolytic event. In vivo mutational analysis demonstrated the lack of any cis-acting sequence elements directing this cleavage within ITS1. PMID- 16701558 TI - Ceramide response post-photodamage is absent after treatment with HA14-1. AB - The oxidative stress induced by photodynamic therapy using the phthalocyanine Pc 4 (PDT) can lead to apoptosis, and is accompanied by photodamage to Bcl-2 and accumulation of de novo ceramide. Similar to PDT, the oxidative stress inducer and Bcl-2 inhibitor HA14-1 triggers apoptosis. To test the specificity of the ceramide response, Jurkat cells were exposed to an equitoxic dose of HA14-1. Unlike PDT, HA14-1 did not induce accumulation of de novo ceramide, although levels of sphingomyelin, phosphatidylserine and phosphatidylethanolamine were below control values after either treatment. In contrast to PDT, (i) the transient inhibition of serine palmitoyltransferase induced by HA14-1 was associated with the initial decrease in de novo ceramide, and (ii) HA14-1 initiated inhibition of sphingomyelin synthase and glucosylceramide synthase did not result in accumulation of de novo ceramide. These results show that the ceramide response to PDT is not induced by another pro-apoptotic stimulus, and may be unique to PDT as described here. PMID- 16701560 TI - Measurement of intracellular IP3 during Ca2+ oscillations in mouse eggs with GFP based FRET probe. AB - Intracellular Ca2+ oscillations in fertilized mammalian eggs, the key signal that stimulates egg activation and early embryonic development, are regulated by inositol 1,4,5-trisphosphate (IP3) signaling pathway. We investigated temporal changes in intracellular IP3 concentration ([IP3]i) in mouse eggs, using a fluorescent probe based on fluorescence resonance energy transfer between two green fluorescent protein variants, during Ca2+ oscillations induced by fertilization or expression of phospholipase Czeta (PLCzeta), an egg-activating sperm factor candidate. Fluorescence measurements suggested the elevation of [IP3]i in fertilized eggs, and the enhancement of PLCzeta-mediated IP3 production by cytoplasmic Ca2+ was observed during Ca2+ oscillations or in response to CaCl2 microinjection. The results supported the view that PLCzeta is the sperm factor to stimulate IP3 pathway, and suggested that high Ca2+ sensitivity of PLCzeta activity and positive feedback from released Ca2+ are important for triggering and maintaining Ca2+ oscillations. PMID- 16701561 TI - Kdr allelic variation in pyrethroid resistant mosquitoes, Culex quinquefasciatus (S.). AB - The loss of target site sensitivity to insecticides resulting from a substitution of leucine to phenylalanine, termed the kdr mutation, in the voltage-gated sodium channel of the insect nervous system is known to be important in insecticide resistance. However, little is known about the molecular basis of the genotype and kdr-mediated resistance phenotype relationship. This study investigated whether the functional polymorphism of the L-to-F kdr mutation that determines resistance phenotype undergoes DNA variation or goes through transcriptional regulatory variation. We detected no correlation for the kdr allele at the genomic DNA level with levels of susceptibility and resistance to insecticide. However, we find a strong correlation between the kdr allelic expression and levels of insecticide resistance and susceptibility through RNA allelic variation and RNA editing. These findings shed new light on the role of transcriptional regulation in the kdr-mediated resistance in mosquitoes and its connection with the genotype-resistance phenotype relationship. PMID- 16701562 TI - Remodeling excitation-contraction coupling of hypertrophied ventricular myocytes is dependent on T-type calcium channels expression. AB - We utilized Wistar rats with monocrotaline (MCT)-induced right ventricular hypertrophy (RVH) in order to evaluate the T-type Ca2+ channel current (ICaT) for myocardial contraction. RT-PCR provides that mRNA for T-type Ca2+ channel alpha1 subunits in hypertrophied myocytes was significantly higher than those in control rats (alpha1G; 264+/-36%, alpha1H; 191+/-34%; P<0.05). By whole-cell patch-clamp study, ICaT was recorded only in hypertrophied myocytes but not in control myocytes. The application of 50 nmol/L nifedipine reduced the twitch tension of the right ventricles equally in the control and RVH rats. On the other hand, 0.5 micromol/L mibefradil, a T-type Ca2+ channel blocker, strongly inhibited the twitch tension of the RVH muscle (control 6.4+/-0.8% vs. RVH 20.0+/-2.3% at 5 Hz; P<0.01). In conclusion, our results indicate the functional expression of T-type Ca2+ channels in the hypertrophied heart and their contribution to the remodeling of excitation-contraction coupling in the cardiac myocyte. PMID- 16701563 TI - Cyclitols protect glutamine synthetase and malate dehydrogenase against heat induced deactivation and thermal denaturation. AB - The accumulation of cyclitols in plants is a widespread response that provides protection against various environmental stresses. The capacity of myo-Inositol, pinitol, quercitol, and other compatible solutes (i.e., sorbitol, proline, and glycinebetaine) to protect proteins against thermally induced denaturation and deactivation was examined. Enzymatic activity measurements of L-glutamine synthetase from Escherichia coli and Hordeum vulgare showed that the presence of cyclitols during heat treatment resulted in a significantly higher percentage of residual activity. CD spectroscopy experiments were used to study thermal stabilities of protein secondary structures upon the addition of myo-Inositol, pinitol, and glucose. 0.4 M myo-Inositol was observed to raise the melting temperature (Tm) of GS from E. coli by 3.9 degrees C and MDH from pig heart by 3.4 degrees C, respectively. Pinitol showed an increase in Tm of MDH by 3.8 degrees C, whereas glucose was not effective. Our results show a great potential of stabilizing proteins by the addition of cyclitols. PMID- 16701564 TI - IGFBP-1 inhibits EGF mitogenic activity in cultured endometrial stromal cells. AB - The properties of the insulin-like growth factor-binding proteins (IGFBP-1 to 6) are not limited to modulation of IGF actions. IGFBP-1, which shares an Arg-Gly Asp (RGD) motif in its C-terminal domain, modulates cell motility by binding to integrin alpha5beta1. The cross-talks between integrins and growth factor receptor signalling pathways are extensively documented, particularly in the case of the epidermal growth factor receptor (EGFR). However, whether IGFBP-1 can modulate growth factor signalling through its interaction with integrin alpha5beta1 has not yet been studied. As EGF is involved in the decidualisation of endometrial stromal cells (ESCs) and as decidualised ESCs are a source of IGFBP-1, we investigated if IGFBP-1 can modulate EGF effects on ESCs. RGD- and IGF-independent inhibition of EGF mitogenic activity and EGFR signalling by IGFBP 1 were demonstrated in ESC primary cultures, A431, cells and in mouse fibroblasts lacking IGF receptors. PMID- 16701567 TI - Animal models simulating anaerobic infections. AB - Animal models simulating human disease have played an important role in our understanding of the pathogenesis and treatment of infections caused by obligately anaerobic bacteria. These models helped document the primary source of such infections as the host's own normal microflora. In addition, the polymicrobial nature of anaerobic infections was documented by using animal models for intraabdominal sepsis. Subsequent studies using animal models have led to an understanding of the nature of the host immune response to abscess causing agents and have been instrumental in defining the molecular basis for the virulence and protection provided by the polysaccharide capsule of Bacteroides fragilis. Animal models have also been important components for studies of toxigenic clostridial diseases, such as antibiotic associated colitis and ulcerative colitis. A discussion of some of these models is provided in this review. PMID- 16701565 TI - Comparative analysis of expression of two p97 homologues in Caenorhabditis elegans. AB - Caenorhabditis elegans possesses two p97/VCP/Cdc48p homologues, named CDC-48.1 (C06A1.1) and CDC-48.2 (C41C4.8), although their expression regulation and functional diversity have not yet been studied. We therefore investigated spatial and temporal expression patterns of two p97 homologues in this study. RT-PCR and Western blot analysis showed that the amount of cdc-48.1 was about twofold of that of cdc-48.2 in adults and that two p97 homologues were induced by ER stress. The amount of cdc-48.1 mRNA did not increase in the cdc-48.2 deletion mutant and vice versa. In situ hybridization showed that two p97 homologues are mainly expressed in germ cells. In vivo expression analysis by using GFP translational fusion constructs revealed that CDC-48.1::GFP was expressed from embryos through to adult worms, while CDC-48.2::GFP was expressed mainly in embryos. These results suggest that the expression of two p97 homologues of C. elegans is differently regulated and independent of each other. PMID- 16701568 TI - A survey of metronidazole and vancomycin resistance in strains of Clostridium difficile isolated in Warsaw, Poland. AB - The drug of choice used to treat Clostridium difficile-associated diarroea (CDAD) are metronidazole and vancomycin. Information about emergence of antimicrobial resistance among C. difficile strains to metronidazole and intermediate resistance to vancomycin in some countries are alarming. This study was performed to determine the susceptibility to metronidazole and vancomycin of 193 C. difficile strains isolated in our diagnostic laboratory between year 1998 and 2003 from patients adults and children suffering from CDAD. Among these strains, 142 produced toxin A and B (TcdA(+)TcdB(+)), 43 only B (TcdA(-)TcdB(+)) and 8 were nontoxigenic. We have not observed any differences in susceptibility to metronidazole and vancomycin between all C. difficile strains under investigation (toxinogenic and non-toxinogenic). Resistance to metronidazole and vancomycin was not observed. PMID- 16701569 TI - Effects of gyrase mutation on the growth kinetics of ciprofloxacin-resistant strains of Clostridium perfringens. AB - To investigate the effect of gyrA mutation on resistance of Clostridium perfringens to fluoroquinolones, a ciprofloxacin-resistant mutant was developed. The mutant had a single substitution in gyrA at position 87 (Asp to Tyr) and no additional mutations in gyrB, parC or parE. The MIC values of gatifloxacin and ciprofloxacin for this strain were 16 and 32-fold higher than those for the wild type, which were 0.125 and 0.250 microg/mL, respectively. The resistant mutant grew equally well in the presence or absence of 5 microg/mL of ciprofloxacin or 1 microg/mL of gatifloxacin and grew to lower cell densities with up to 30 microg/mL of ciprofloxacin or 5 microg/mL of gatifloxacin. Higher concentrations of fluoroquinolones resulted in increases in the time required to reach the end of the exponential phase and in lower cell densities at the end. The efflux pump inhibitor reserpine did not affect susceptibility to fluoroquinolones. The substitution of Asp 87 to Tyr in gyrA may have protected C. perfringens from low concentrations of ciprofloxacin and gatifloxacin and enabled survival and growth at higher concentrations. PMID- 16701566 TI - Evidence for post-transcriptional regulation of Na,K-ATPase by prostaglandin E1. AB - The stimulatory effect of PGE1 on the activity of the Na,K-ATPase in MDCK cells is associated with an increase in the rate of transcription of the Na,K-ATPase beta1 subunit gene and an increase in the rate of biosynthesis of the Na,K-ATPase [M.L. Taub, Y. Wang, I.S. Yang, P. Fiorella, S.M. Lee, Regulation of the Na,K ATPase activity of Madin-Darby canine kidney cells in defined medium by prostaglandin E1 and 8-bromocyclic AMP, J. Cell. Physiol. 151 (1992) 337-346]. In order to further define the molecular mechanisms, transient transfection and biosynthesis studies were conducted with dibutyryl cAMP resistant (DBr) MDCK cells, defective in cAMP dependent protein kinase, and PGE1 independent (PGE1 Ind) MDCK cells with elevated intracellular cAMP. Transient transfection studies with the human Na,K-ATPase beta1 promoter/luciferase construct, pHbeta1-1141 Luc [J. Feng, J. Orlowski, J.B. Lingrel, Identification of a functional thyroid hormone response element in the upstream flanking region of the human Na,K-ATPase beta 1 gene, Nucleic Acids Res. 21 (1993) 2619-2626], showed that the stimulatory effect of PGE1 and 8Br-cAMP on beta1 subunit gene transcription is retained in the DBr and PGE1 independent variants. However, the stimulatory effect of PGE1 and 8Br-cAMP on Na,K-ATPase biosynthesis was lost in DBr (unlike PGE1 Ind) variants. These results can be explained by a defect in post-transcriptional regulation. PMID- 16701570 TI - Prokaryote diversity in the rumen of yak (Bos grunniens) and Jinnan cattle (Bos taurus) estimated by 16S rDNA homology analyses. AB - Prokaryote diversity in the rumen of yak (Bos grunniens) and Jinnan cattle (Bos taurus) was estimated by 16S rDNA homology analysis. Two rumen 16S rDNA libraries were constructed. Of the 194 clones in the library of yak rumen, the sequences were mainly clustered to two phyla, low G+C Gram-positive bacteria (LGCGPB, 54.12% total clones) and Bacteroidetes (30.93%), respectively. While in the 197 clone-library of the cattle rumen, the sequences were mainly related to three phyla, Bacteroidetes (39.59%), gamma-Proteobacteria (26.9%) and LGCGPB (22.34%), respectively. The sequence analysis indicated that more than half of the species harbored in yak rumen belonged to the not-yet-cultured groups at <90% 16S rDNA similarity levels with cultured species, while 36% 16S rDNA sequences amplified from the rumen of Jinnan cattle fell in these catalogues. By comparing the uncultured sequences in yak rumen with those in Jinnan cattle and cow, the former formed distinct clusters loosely related to the later, implying that yak rumen could harbor some special prokaryote phyla. 10.8% sequences retrieved in yak rumen were related to the known rumen fibrolytic bacterial species; however none was related to the known amylolysis species. While 4% and 17.8% sequences retrieved from Jinnan cattle rumen were related to cultured fibrolytic and amylolysis species, respectively. The bacterial structures seemed to be in accordance with the feed of the two kinds of animals. In both rumens, retrieved methanogenic Archaea-related 16S rDNA sequences were at an unreasonable low level; in addition, none sequence was related to Ruminococcus albus, a classical rumen fibrolytic species. The reason can be due to the experimental biases. PMID- 16701571 TI - Immobilization of anaerobic bacteria on rubberized-coir for psychrophilic digestion of night soil. AB - Low-ambient temperatures, <30 degrees C, are known to cause drastic reduction in the efficiency of anaerobic biodigesters due to low-growth rate of the constituent bacterial consortium. Immobilization of anaerobic bacteria has been attempted in the biodigester operating at 10 degrees C. Various matrices were screened and evaluated for the immobilization of bacteria in digesters. Anaerobic digestion of night soil was carried out with hydraulic retention time in the range of 9-18 days. Among the tested matrices, rubberized-coir was found to be the most useful at 10 degrees C with optimum hydraulic retention time of 15 days. Optimum amount of coir was found as 25 g/L of the working volume of biodigesters. Immobilization of bacteria on the coir was observed by scanning electron microscopy and fluorescent microscopy. The study indicates that rubberized-coir can be utilized to increase biodegradation of night soil at higher organic loading. Another advantage of using this matrix is that it is renewable and easily available in comparison to other synthetic polymeric matrices. PMID- 16701572 TI - Feruloyl oligosaccharides stimulate the growth of Bifidobacterium bifidum. AB - Insoluble dietary fiber from wheat bran contains some feruloyl groups linked to the arabinose residues in the cell wall arabinoxylan. Treatment of wheat bran insoluble dietary fiber with xylanase from Bacillus subtilis yielded feruloyl oligosacchairdes, which were purified with Amberlite XAD-2. Saponification of the feruloyl oligosaccharides released ferulic acid and arabinoxylan oligosaccharides which consist of arabinose and xylose. The effect of the feruloyl oligosacchairdes on the growth of Bifidobacterium bifidum F-35 was investigated in vitro. The B. bifidum produced acid when cultivated anaerobically in TPY broth with 0.5% feruloyl oligosacchairdes as the carbohydrate source. The biomass yield of the B. bifidum increased with increasing the concentration of feruloyl oligosaccharides in TPY broth. The maximum cell growth was increased by 50% in TPY broth supplemented with 0.1% feruloyl oligosaccharides compared to TPY broth. These results indicated that the growth of B. bifidum F-35 was promoted by the feruloyl oligosaccharides from wheat bran insoluble dietary fiber, and not suppressed by the ferulic acid moiety of them. PMID- 16701573 TI - Characterization of Actinomyces species isolated from failed dental implant fixtures. AB - In the oral cavity, Actinomyces form a fundamental component of the indigenous microflora, being among initial colonizers in polymicrobial biofilms. However, some differences may exist between different species in terms of their attachment not only to teeth but also to biomaterials. In this study we investigated the distribution of Actinomyces in 33 dental implant fixtures explanted from 17 patients. The identification was based on comprehensive biochemical testing and gas-liquid chromatography and when needed, 16S rRNA sequencing. Actinomyces was the most prevalent bacterial genus in these failed implants, colonizing 31/33 (94%) of the fixtures. Proportions of Actinomyces growth of the total bacterial growth in the Actinomyces-positive fixtures varied from 0.01% up to 75%. A. odontolyticus was the most common Actinomyces finding, present in 26/31 (84%) Actinomyces-positive fixtures. Actinomyces naeslundii and A. viscosus were both detected in 10/31 (32%) and A. israelii in 7/31 (23%) fixtures. Other Actinomyces species, including A. georgiae, A. gerencseriae and A. graevenitzii, were detected less frequently. Our results suggest that Actinomyces species are frequent colonizers on failed implant surfaces, where A. odontolyticus was the far most prominent Actinomyces species. PMID- 16701574 TI - Fusobacterium necrophorum infections in animals: pathogenesis and pathogenic mechanisms. AB - Fusobacterium necrophorum, a Gram-negative, non-spore-forming anaerobe, is a normal inhabitant of the alimentary tract of animals and humans. Two subspecies of F. necrophorum, subsp. necrophorum (biotype A) and subsp. funduliforme (biotype B), have been recognized, that differ morphologically, biochemically, and biologically. The subsp. necrophorum is more virulent and is isolated more frequently from infections than the subsp. funduliforme. The organism is an opportunistic pathogen that causes numerous necrotic conditions (necrobacillosis), either specific or non-specific infections, in a variety of animals. Of these, bovine liver abscesses and foot rot are of significant concern to the cattle industry. Liver abscesses arise with the organisms that inhabit the rumen gaining entry into the portal circulation, and are often secondary to ruminal acidosis and rumenitis complex in grain-fed cattle. Foot rot is the major cause of lameness in dairy and beef cattle. The pathogenic mechanism of F. necrophorum is complex and not well defined. Several toxins or secreted products, such as leukotoxin, endotoxin, hemolysin, hemagglutinin, proteases, and adhesin, etc., have been implicated as virulence factors. The major virulence factor appears to be leukotoxin, a secreted protein of high molecular weight, active specifically against leukocytes from ruminants. The complete nucleotide sequence of the leukotoxin operon of F. necrophorum has been determined. The operon consists of three genes (lktBAC) of which the second gene (lktA) is the leukotoxin structural gene. The leukotoxin appears to be a novel protein and does not share sequence similarity with any other leukotoxin. F. necrophorum is also a human pathogen and the human strains appear to be different from the strains involved in animal infections. PMID- 16701575 TI - Neurochemically defined cell columns in the nucleus prepositus hypoglossi of the cat and monkey. AB - Many studies have shown that the nucleus prepositus hypoglossi (PH) participates with the vestibular nuclear complex, the cerebellum and the oculomotor nuclei in the control of eye movements. We have looked at the neurochemical organization of PH in the cat and monkey using a recently developed antibody, 8B3, that recognizes a chondroitin sulfate proteoglycan. In the cat, immunoreactivity to 8B3 labels a set of cells in PH. On frontal sections, these cells form a cluster that is seen over the entire anterior-posterior (A-P) extent of PH, but the number of cells in the cluster changes with A-P level. Earlier studies have identified an A-P cell column in PH of the cat whose neurons synthesize nitric oxide. We have used both single- and double-label protocols to investigate the relation between the two cell groups. Single-label studies show spatial overlap but that the cells immunoreactive to nitric oxide synthase (nNOS) are more numerous than cells immunoreactive to 8B3. Double-label studies show that all cells immunoreactive to 8B3 were also immunoreactive to nNOS, but, as suggested by the single-label data, there are many nNOS-immunoreactive cells not immunoreactive to 8B3. Populations of 8B3 and nNOS-immunoreactive cells are also found in PH of squirrel and macaque monkeys. The results suggest that nNOS immunoreactive cells in PH may consist of two functionally different populations. PMID- 16701577 TI - Distribution of inducible nitric oxide synthase and cell proliferation in rat brain after transient middle cerebral artery occlusion. AB - Nitric oxide (NO) can be neuroprotective or neurotoxic during cerebral ischemia, depending on the NO synthase (NOS) isoform involved. In addition to neurotoxic effect in ischemic brain, inducible NOS (iNOS) also adversely affect ischemic outcome by blocking neurogenesis. In the present study, therefore, we studied the chronological and spatial change of the distribution of iNOS and cell proliferation in subventricular zone (SVZ) after transient focal cerebral ischemia. After 90 min of transient middle cerebral artery occlusion (tMCAO), iNOS-positive cells decreased in the ischemic core at 1 to 21 days, and increased in the ipsilateral periischemic area at 1 and 3 days. 5-Bromodeoxyuridine (BrdU) positive cells appeared in the ischemic core at 3 to 21 days, appeared in the periischemic area at 3 and 7 days, and increased in the ipsilateral SVZ at 7 days. ED-1-positive cells appeared in the ischemic core at 3 to 21 days, and some of them were double positive with BrdU or iNOS, but the majority were BrdU negative. The present study suggests that astrocytes are born within the periischemic area at early stage after tMCAO and migrate from SVZ into periischemic area at later stage, and that time-dependent and spatial changes of iNOS expression may be involved in the proliferation and differentiation of adult neurogenesis after focal cerebral ischemia. PMID- 16701576 TI - Multiple receptors mediate the trophic effects of serotonin on ventroposterior thalamic neurons in vitro. AB - Serotonin (5-HT) exerts prominent morphogenetic roles during development. For example, somatosensory cortical barrel formation is altered in mouse models characterized by excessive extracellular 5-HT, suggesting that 5-HT affects development of thalamic afferents and/or neocortical target regions. The present study assessed 5-HT effects in primary cultures of fetal ventroposterior thalamic (VPT) neurons. 5-HT produces concentration-dependent trophic effects, with impressive 59% and 106% peak increases in total neurite length and number of branching points, respectively, at a dose of 30 microM 5-HT. The exposure of VPT neurons to specific 5-HT receptor agonists 8-OH-DPAT (5-HT(1A)), CGS-12066A (5 HT(1B)), DOI (5-HT(2A/2C)), and m-CPBG (5-HT(3)), enhances primary neurite length and number of branching points with rank-order potency 5-HT(1B) > 5-HT(2A/2C) = 5 HT(3) > 5-HT(1A) = vehicle. Trophic 5-HT effects on embryonic VPT neurons are thus much more prominent than previously reported, and can be mediated by multiple 5-HT receptor subtypes. PMID- 16701579 TI - A dynamic partnership: celebrating our gut flora. AB - Emerging data indicate that humans enjoy health through a productive collaboration with their colonizing flora, the majority of whom reside in the colon. This minireview provides a perspective on recent data and the exciting scientific challenges ahead. PMID- 16701578 TI - 5-Aminolevulinate and 4, 5-dioxovalerate ions decrease GABA(A) receptor density in neuronal cells, synaptosomes and rat brain. AB - Porphyrias are heme-associated metabolic disorders such as intermittent acute porphyria (IAP) and lead poisoning, where 5-aminolevulinate (ALA) accumulates. Effects of ALA on the CNS have been explained by ALA binding to GABA(A) receptors, followed by receptor lesions from oxyradicals and 4, 5-dioxovalerate (DOVA) generated from metal-catalyzed ALA oxidation by oxygen. We have characterized the effects of ALA and DOVA on GABA(A) receptor density in synaptosomes and neurons in vitro and also in brains of rats treated with ALA or succinylacetone methyl ester (SAME), a tyrosine catabolite derivative able to induce ALA accumulation. Radiolabeling assays revealed that following exposure to DOVA the concentration of synaptosomal GABAergic sites decreased by approximately 50%. Pretreatment with DOVA resulted in less GABA(A) receptor density in P19 and WERI cells and altered cell morphology. Furthermore, exposure to DOVA also induced a 5-fold increase in WERI cell mortality rate. Treatment with ALA resulted in loss of neuronal morphology and decrease of GABA(A) density in P19 neuronal cells. ALA and SAME treatment diminished the density of GABAergic receptors in the habenular complex and the parabigeminal nucleus of rat brain as studied by immunohistochemical procedures. Our results strongly suggest that ALA- and DOVA-promoted damage to GABA(A) receptors may contribute to the neurological manifestations of AIP and plumbism. PMID- 16701580 TI - The role of anaerobic bacteria in acute and chronic mastoiditis. AB - Mastoiditis (M) is the most common intratemporal complication of otitis media. The incidence of M has decreased since the advent of antimicrobial agents. In the last decade, however, there has been a marked increased in the incidence of acute M in several communities, sometimes in association with the growing resistance of pneumococci. Streptococcus pneumoniae, Streptococcus pyogenes, Staphylococcus aureus and Haemophillus influenzae are the most common organisms recovered in acute M. Several recent studies demonstrated the predominance of Pseudomonas aeruginosa in this infection. However, because P. aeruginosa colonizes the ear canal it can contaminate specimens obtained through the non-sterile ear canal. P. aeruginosa, Enterobacteriaceae, S. aureus and anaerobic bacteria are the most common isolates in chronic M. Anaerobes predominate in studies where adequate methods for their isolation are employed. Management of uncomplicated M requires the administration of parenteral antimicrobial therapy and myringotomy with or without tympanostomy tube placement. If no improvement occurs within 48 h, surgical intervention and drainage may be necessary. The procedure generally employed is simple mastoidectomy, and tympanostomy tube placement. Radical mastoidectomy is performed only if there is no improvement after simple mastoidectomy. PMID- 16701581 TI - Clostridium difficile in emergency room. AB - Clostridium difficile strains are known as etiological agents of pseudomembranous colitis (PMC), antibiotic-associated diarrhea (AAC) and colitis (AAC) and hospital-acquired infections. The aim of this study was to determine the frequency of C. difficile infection among patients in the emergency room and to compare isolated strains by phenotypic and genotypic characteristics. During a period of 11 months, 56 stool samples taken from diarrheic patients hospitalized in the emergency room of the Medical Center UC Davis and 14 environmental samples were cultured for isolation of C. difficile strains. Eighteen C. difficile strains were isolated from stool samples cultured on selective TCCCA plates and 5 strains from environmental samples using Rodac plates. Eleven toxigenic (TcdA+/TcdB+), 6 non-toxigenic (TcdA-/TcdB-) and unique toxin A-negative/toxin B positive (TcdA-/TcdB+) C. difficile strains were detected among patients' isolates and 3 toxigenic and 2 non-toxigenic strains-among environmental samples. The majority of C. difficile-positive patients were treated previously by antibiotics. Four strains isolated from patients' fecal samples and one strain isolated from the environment demonstrated high-level resistance to erythromycin and clindamycin (MIC >256mug/mL). The results obtained by AP-PCR and PCR ribotyping revealed genetic heterogeneity among the strains isolated from patients' fecal samples. However, similarity was observed among environmental strains and strains isolated from patients' fecal samples. Considering the importance of emergency room patients as a potential source of C. difficile strains, it appears to be important examine these patients for C. difficile before transfer to the other hospital units. PMID- 16701582 TI - Multiple-locus variable-number tandem repeat analysis for strain typing of Clostridium perfringens. AB - Clostridium perfringens is ubiquitous in the environment and causes diseases in man and animals, with syndromes ranging from enteritis, enterotoxemia, and sudden death to food poisoning and gas gangrene. Understanding the epidemiology of these infections and of the evolution of virulence in C. perfringens necessitate an efficient, time and cost effective strain typing method. Multiple-locus variable number tandem repeat analysis (MLVA) has been applied to typing of other pathogens and we describe here the development of a MLVA scheme for C. perfringens. We characterized five variable tandem repeat (VNTR) loci, four of which are contained within protein encoding genes and screened 112 C. perfringens isolates to evaluate typability, reproducibility, and discriminatory power of the scheme. All the isolates were assigned a MLVA genotype and the technique has excellent reproducibility, with a numerical index of discrimination for the five VNTR loci of 0.995. Thus MLVA is an efficient tool for C. perfringens strain typing, and being PCR based makes it rapid, easy, and cost effective. In addition, it can be employed in epidemiological, ecological, and evolutionary investigations of the organism. PMID- 16701583 TI - Matrix bound phosphine formation and depletion in eutrophic lake sediment fermentation-simulation of different environmental factors. AB - Closed anaerobic batch-fermentation of eutrophic lake sediment samples was performed under variation of four environmental fermentation factors (pH, temperature, water/sediment ratio and disturbance) to learn how the quantity of phosphine will change and if the quantity of phosphine can increase. The fermentation conditions where matrix bound phosphine (MBP) increased (doubled from 3193+/-520 to about 7982+/-1003 ng/kg) were: a pH of 8 and of 10 (as compared to 1, 2, 4, 6, 12), a temperature of 20 and 30 degrees C (as compared to 4 and 40 degrees C), a water/sediment ratio of 3:1 (as compared to 1:1, 2:1, 5:1) and a disturbance of 100 r/min (as compared to 0 r/min), respectively. Although, over the full time course of fermentation, the balance of phosphine production became negative again or did remain almost unchanged under most conditions. A pH of 1 or disturbance of 150 r/min was significant factors to decrease phosphine over the long term. Free phosphine had been detected but was of minor importance (in the order of 60.9+/-10.1 ng/m(3)). Overall, the fermentation conditions which had been most favorable for microbial life (moderate temperature (20 and 30 degrees C) and pH 8) were also most favorable for a positive phosphine balance. This is an indication, but no biochemical proof that a natural (biogenic, microbial, biochemical) NET PRODUCTION of phosphine or DE NOVO PRODUCTION of phosphine has occurred. MBP concentrations in lake sediments were discussed as to be strongly dependent on a balance of natural generation and depletion processes, dependent of the simulated parameters. PMID- 16701584 TI - Occurrence of restriction-modification systems in ruminal butyrate-producing bacteria. AB - Thirty-five strains of ruminal bacteria belonging to the former Butyrivibrio fibrisolvens species were screened for the presence of site-specific restriction endonuclease and modification methyltransferase activities. Seven strains possessed endonuclease activities detectable in crude cell extracts. The recognition sequences and optimal reaction conditions for seven of them were determined. Five enzymes were found to be isoschizomers of type II endonucleases (EcoRV, NsiI, AseI (2x) and SauI), one was type IIS (FokI) and two remained unknown. The optimal reaction buffer was found to be a low ionic strength buffer and all enzymes possessed sufficient activity at 39 degrees C. The presence of DNA modification among all strains was also determined. Most of the methylation activities correlated with restriction activities, yet some strains possessed unaccompanied modification methyltransferases. PMID- 16701585 TI - Properties of hemagglutination by Prevotella melaninogenica. AB - Although Prevotella melaninogenica belongs to the commensal oral microbiota, some strains possess putative virulence factors. For example, we have previously described fimbriated, hemagglutinating strains of P. melaninogenica, isolated from patients with periodontal disease. The aim of this investigation was to compare some chemical and physical properties of hemagglutination (HA) of P. melaninogenica with those of other pigmented gram-negative anaerobes. HA of 13 P. melaninogenica strains proved to be considerably weaker than that of the major periodontal pathogen, Porphyromonas gingivalis. Vigorous shaking reduced HA of shaken cells but the shaken supernatant had the same hemagglutinating activity as non-shaken cells. The hemagglutinating agent on P. melaninogenica seemed to be a protein, which can be separated from the cell and binds to lactose-, galactose-, and raffinose-containing carbohydrates on the erythrocytes. Adherence to epithelial cells did not differ significantly between the hemagglutinating and non-hemagglutinating strains of P. melaninogenica. Although P. melaninogenica is able to agglutinate erythrocytes, this potential virulence factor is of a considerably lower magnitude than that of major periodontal pathogens. PMID- 16701586 TI - Clostridial abomasitis in calves: case report and review of the literature. AB - Infections by Clostridium perfringens type A are perhaps the most common causes of clostridial hemorrhagic enteritis in neonatal ruminants. Affected calves exhibit tympany, hemorrhagic abomasitis, and abomasal ulceration. Gram-positive bacilli are often found on affected mucosa and in submucosa. Aspects of etiology beyond the infecting organism are little understood, but probably include dietary issues, perhaps relating to overfeeding, feeding of barely thawed or contaminated colostrum, or conditions which effect decreased gut motility. Fatal hemorrhagic enteritis in a cloned gaur calf is illustrative of the syndrome. The calf developed pasty yellow and bloody diarrhea, and the abdomen became distended and painful. In spite of intensive therapy, the calf died approximately 48 h after birth. At necropsy, the distended abomasum contained clotted milk and bloody fluid, and the abomasal and omasal walls were thickened and hemorrhagic. The proximal duodenum was hemorrhagic and emphysematous, and microscopic examination revealed Gram-positive rods in association with acute, necrotizing, hemorrhagic mucosal inflammation. Isolates of C. perfringens from this calf were PCR positive for cpb2, the gene encoding beta2 toxin. This finding is of unknown significance; only 14.3% (8/56) of isolates from other calves with the syndrome have been cpb2 positive, and only 50% of cpb2 positive bovine isolates express CPB2. The most prominent needs to further our understanding of this problem are consistent experimental reproduction of the disease, elucidation of virulence attributes, and development and application of prevention and control strategies. PMID- 16701587 TI - Bacteroides species produce Vibrio harveyi autoinducer 2-related molecules. AB - Quorum sensing is a density-dependent gene regulation mechanism that has been described in many bacterial species in the last decades. Bacteria that use quorum sensing as part of their gene regulation circuits produce molecules called autoinducers that accumulate in the environment and activate target genes in a quorum-dependent way. Some specific clues led us to hypothesize that Bacteroides species can produce autoinducers and possess a quorum sensing system. First, Bacteroides are anaerobic bacteria that are frequently involved in polymicrobial infections. These infections often involve Pseudomonas aeruginosa and Staphylococcus aureus, two of the best understood examples of bacteria that employ quorum sensing systems as part of their pathogenesis. Also, studies have detected the presence of a quorum sensing gene involved in the production of autoinducers in Porphyromonas gingivalis, a species closely related to the Bacteroides genus. These and other evidences prompted us to investigate if Bacteroides strains could produce autoinducer molecules that could be detected by a Vibrio harveyi reporter system. In this paper, we show that supernatants of B. fragilis, B. vulgatus and B. distasonis strains are able to stimulate the V. harveyi quorum sensing system 2. Also, we were able to demonstrate that the stimulation detected is due to the production of autoinducer molecules and not the growth of reporter strains after addition of supernatant. Moreover, the phenomenon observed does not seem to represent the degradation of repressors possibly present in the culture medium used. We could also amplify bands from some of the strains tested using primers designed to the luxS gene of Escherichia coli. Altogether, our results show that B. fragilis, B. vulgatus and B. distasonis (but possibly some other species) can produce V. harveyi autoinducer 2 related molecules. However, the role of such molecules in the biology of these organisms remains unknown. PMID- 16701589 TI - Infection and mental illness: do bugs make us batty? PMID- 16701588 TI - Convenient synthesis of a sialylglycopeptide-thioester having an intact and homogeneous complex-type disialyl-oligosaccharide. AB - Access to glycopeptides with C-terminal thioester functionality is essential for the synthesis of large glycopeptides and glycoproteins through the use of native chemical ligation. Toward that end, we have developed a concise method for the synthesis of a glycopeptide thioester having an intact complex-type dibranched disialyl-oligosaccharide. The synthesis employed a coupling reaction between benzylthiol and a free carboxylic acid at the C-terminus of a glycopeptide in which the peptide side chains are protected. After construction of glycopeptide on the HMPB-PEGA resin through the Fmoc-strategy, the protected glycopeptide was released upon treatment with acetic acid/trifluoroethanol and then the C-terminal carboxylic acid was coupled with benzylthiol at -20 degrees C in DMF. For this coupling, PyBOP/DIPEA was found to be the best for the formation of the thioester, while avoiding racemization. Finally, the protecting groups were removed in good yield with 95% TFA, thus affording a glycopeptide-thioester having an intact and homogeneous complex-type disialyl-oligosaccharide. PMID- 16701590 TI - The association of anaerobic bacteria with infectious mononucleosis. AB - This review summarizes the information that supports the potential importance of anaerobic bacteria in the pharyngo-tonsillitis (PT) associated with infectious mononucleosis (IM). Evidence supportive of the potential of involvement of bacteria in IM was provided by several studies that showed increased recovery of bacteria on the tonsillar surfaces in IM. Several studies demonstrated that metronidazole therapy alleviated the clinical symptoms of tonsillar hypertrophy and shortened the duration of fever in IM. Metronidazole has no antimicrobial activity against aerobic bacteria and is only effective against anaerobic bacteria. A possible mechanism of its action is the suppression of the oral anaerobic flora that might contribute to the inflammatory process induced by the Epstein-Barr virus. This explanation is supported by the increased recovery of Prevotella intermedia and Fusobacterium nucleatum from the tonsillar surfaces during the acute phases of IM and an immune response against these organisms by patients with IM. Although more studies are needed, these findings support the possible pathogenicity of Gram-negative anaerobic bacteria in the PT associated with IM. PMID- 16701591 TI - FTIR-spectroscopic studies of the fine structure of nitrocellulose treated by Desulfovibrio desulfuricans. AB - Most studies have concluded that nitrocellulose (NC) with high degree of nitrogen content is resistant to biodegradation. Our results demonstrated that NC (>11%N) does undergo biotransformation in the presence of sulfate-reducing bacteria Desulfovibrio desulfuricans 1388. FTIR analyses indicated that the substitution of nitro groups for OH(-) groups took place. The spectrum of precipitate obtained after acetone extraction of NC resembled mainly the spectrum of native cellulose. Thus the synthetic unbiodegradable polymer was transformed to the natural compound accessible for microorganisms. PMID- 16701593 TI - Prevalence of enterotoxigenic Bacteroides fragilis in patients with diarrhea: a controlled study. AB - In this age matched controlled study performed in Malatya, a city in east region of Turkey, enterotoxigenic Bacteroides fragilis (ETBF) was investigated in stool specimens obtained from children and adults with and without diarrhea. A nested polymerase chain reaction (PCR) method was used to detect the enterotoxin gene of B. fragilis in a total of 418 stool samples, including 221 samples from 117 children (aged 0-16 years) and 104 adults (aged >16 years) with diarrhea, and 197 samples from 102 children and 95 adults as control group that was the same age group with those having diarrhea. ETBF was detected in 13 of 117 diarrheal children (11.1%) and 8 of 102 control children (7.8%) (P>0.05). In children aged 1-5 years, the rate of ETBF was significantly higher in patients than in controls (25% versus 9.5%, respectively; P<0.05). On the other hand ETBF was detected similar rates (2.2% and 2.4%, respectively) in children younger than 1 year in both patients and controls. ETBF positivity was not significantly difference between patient and control groups who were older than 5 years of age and adults. The frequency of ETBF in the controls was slightly higher in older persons than in younger ones; however, it was not significant. The rate of ETBF as the only enteropathogen in the patients with ETBF was significantly higher than in controls with ETBF (88% versus 39%, respectively; P<0.02). We found that in east region of Turkey, the prevalence of ETBF was higher in the childhood diarrhea, particularly in aged 1-5. As the only enteropathogen, ETBF may play an important role in diarrheal diseases. Persons after 6 years old can be carrier for ETBF regardless diarrhea. PMID- 16701592 TI - In vitro activities of tinidazole and metronidazole against Clostridium difficile, Prevotella bivia and Bacteroides fragilis. AB - Tinidazole, a 5-nitroimidazole similar to metronidazole, was studied against 40 Clostridium difficile, 10 Prevotella bivia and 11 Bacteroides fragilis clinical isolates. The geometric mean MICs of tinidazole and metronidazole were, respectively: C. difficile, 0.31 and 0.28 microg/mL; P. bivia, 2.33 and 1.52 microg/mL; B. fragilis, 0.5 and 0.71 microg/mL. PMID- 16701594 TI - Diversity of mesophilic clostridia in Costa Rican soils. AB - Costa Rica is located in the Tropic, one of the most biologically diverse regions of the world; its soil is an important epicenter of biodiversity and Clostridium spp. are among the most frequent bacteria. The diversity of clostridia in Costa Rican soils and its possible association with geographic zone, pH or type of soil was studied in 117 soil samples: 18 from the Atlantic Zone, 30 from the Central Plateau, 30 from the Dry Pacific, 13 from the North Zone, and 26 from the South Pacific. The pH and the mesophilic clostridia species were determined for each sample. For bacterial isolation, a selective methodology for spores and pre reduced anaerobically sterilized media were used. A total of 1945 strains of clostridia were isolated, 98% were identified and corresponded to 54 species; the most frequent species were C. subterminale (56%), C. oceanicum (51%), C. bifermentans and C. glycolicum (50%, each), C. sporogenes (49%), and C. sordellii (42%). An average of 7.1 species per sample was obtained; the Atlantic Zone showed the greatest diversity: 8.6 species per sample and a total of 45 species. Except for C. chauvoei, all described toxigenic clostridia species were isolated; C. sordellii (42%) and C. perfringens (38%) were the most frequent. No statistical relation could be established between geographic zone or type of soil and any species, showing that clostridia had a high adaptation capability to grow in different soil conditions; only some clostridia were isolated from very acidic samples while others from soils with a wide range of pH. In general, a uniform distribution of most species and a high variety of clostridia in Costa Rican soils were observed, in agreement with the high biodiversity described for other living beings in this country. PMID- 16701595 TI - Prevalence and characterization of Clostridium perfringens from spices in Argentina. AB - Spices can present high microbial counts and Clostridium perfringens, Bacillus cereus, Salmonella and Shigella, among others have been isolated from spices. C. perfringens is an important pathogen agent causing, among other diseases, enteritis in humans caused by C. perfringens enterotoxin (CPE) which causes human food poisoning and enterotoxemia in domestic animals. The aims of the present work were (i) to establish the hygienic sanitary quality of some spices in San Luis, Argentina; (ii) to determine the presence of C. perfringens in these spices by means of the most probable number (MPN) and count on plate methods; (iii) to characterize the enterotoxigenic strains of C. perfringens by PCR and immunological methods such as reverse passive latex agglutination (RPLA) and (iv) to type by PCR C. perfringens strains isolated. A total of 115 samples of spices, 67 of which were purchased in local retail stores and 48 domestically collected were analysed. Total aerobe counts on tryptone glucose yeast extract agar medium of the 115 samples were between <10 and 10(6) CFU/g. The colifecal counts using Mac Conkey broth of the 115 samples were <4-10(3)CFU/g, with 28 samples (24.34%) exceeding the limit established by the Spanish Alimentary Code (10 CFU/g) while 2 samples (1.73%) had a sulfite reducing anaerobe load above standard limits. A total of 14 C. perfringens strains (12.17%) were isolated and characterized from 115 samples by the standard biochemical tests. Four of which (28.60%) turned out to be enterotoxigenic by PCR and RPLA. In order to type C. perfringens strains based on their main toxins, the 14 strains were analysed by PCR. All strains belonged to type A. All RPLA positive strains were cpe(+) by PCR. The percentage of enterotoxigenic strains was more elevated that those reported in other studies for this type of sample. These results indicate that sanitary conditions in different production stages of species must be improved to reduce health hazards. The high percentage (24.34%) of samples with colifecal values above standard limits is an indication of deficient sanitary conditions. These results suggest the need to provide legislation on the sanitary and hygienic quality of spices in our country. PMID- 16701596 TI - Carbon monoxide inhibits superoxide dismutase and stimulates reactive oxygen species production by Desulfovibrio desulfuricans 1388. AB - The hypothesis that oxidative stress characterized by enhanced superoxide generation underlies the toxicity of some factors to living organisms has been investigated. It is shown that CO (5-6% in gas phase) changed some growth parameters (mu, t(d)) of the sulfate-reducing bacterium Desulfovibrio desulfuricans 1388. Enhanced O(2)(-) generation registered by EPR spectroscopy and adrenochrome method was observed when cells were incubated under CO. The SOD activity in cells from the exponential growth phase growing under CO was decreased 1.5-fold compared with the control cells growing under Ar. SOD activities in cells from the stationary growth phase growing with or without CO were comparable. The results support the concept that CO toxicity for sulfate reducing bacteria is an oxidative stress that arises in cells oxidizing CO to CO(2). PMID- 16701597 TI - Desulfovibrio aerotolerans sp. nov., an oxygen tolerant sulphate-reducing bacterium isolated from activated sludge. AB - A new mesophilic sulphate-reducing bacterium, designated strain DvO5(T) (T=type strain), was isolated from the outermost sulphate reduction-positive most probable-number tube (10(-6) dilution) of an activated sludge sample, which had been oxygenated at 100% air saturation for 120 h. The motile, Gram-negative, curved 1 by 2-5 microm and non-spore-forming cells of strain DvO5(T) existed singly or in chains. Strain DvO5(T) grew optimally at 29 degrees C, pH 6.9 and 0.05% (w/v) NaCl in a medium containing lactate, sulphate and yeast extract. Sulphite, thiosulphate and elemental sulphur also served as electron acceptors whereas nitrate, nitrite or ferric iron were not reduced. Lactate, pyruvate, H(2) (with acetate as carbon source), ethanol and glycerol efficiently supported growth as electron donors. Pyruvate and malate were fermented. Strain DvO5(T) reduced oxygen by oxidising endogenous polyglucose at rates ranging from 0.4 to 6.0 nmol O(2)/mg protein min depending on the oxygen concentration, the highest rates being observed at atmospheric oxygen saturation. The G+C content of the DNA was 57.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain DvO5(T) was a member of the genus Desulfovibrio with D. magneticus (98.2% 16S rRNA gene sequence similarity) and D. burkinensis (97.5% 16S rRNA gene sequence similarity) being its closest relatives among validly described species. A similar phylogenetic affiliation was obtained by sequence analyses of the genes encoding the alpha and the beta subunit of dissimilatory sulphite reductase (dsrAB) as well as the alpha subunit of adenosine-5'-phosphosulphate reductase (apsA) of strain DvO5(T). On the basis of genotypic and phenotypic characteristics, strain DvO5(T) (DSM 16695(T), JCM 12613(T)) is proposed as the type strain of a new species, Desulfovibrio aerotolerans sp. nov. PMID- 16701598 TI - Leptotrichia buccalis bacteremia in two patients with acute myelogenous leukemia. AB - Leptotrichia buccalis is rarely implicated in systemic disease. We report two patients with clinically significant L. buccalis bacteremia which developed during the neutropenia secondary to chemotherapy. Based upon our experience, L. buccalis bacteremia should be considered in certain high-risk immunocompromised patients with mucositis and/or gingivitis. PMID- 16701599 TI - A selected history and future of immunoassay development and applications in clinical chemistry. AB - BACKGROUND: The first immunoassay was described by Berson and Yalow in 1959. Their work resulted in their receipt of the Nobel Prize in Medicine in 1977. Since this introduction, immunoassays have evolved considerably. METHODS: There have been several milestones that have led to the proliferation of modern immunoassays. The development of monoclonal antibodies from mouse hydridoma cells by Millstein and Kohler (Nobel Prize in 1984) enabled the production of high quantities of antibodies with well characterized epitope specificity. The first homogenous immunoassay (no separation step required) was the Enzyme Multiplied Immunoassay Technique (EMIT), which enabled adaptation of this assay onto automated chemistry platforms. EMIT was also one of the first immunoassay that made use of non-isotopic labels. Other non-isotopic labels became available such as chemiluminescence to improve the analytical sensitivity of immunoassays. The advantages of high-sensitivity immunoassays have created expanded diagnostic roles for some existing assays such as thyroid stimulating hormone for hyperthyroidism, C-reactive protein for cardiovascular risk assessment, and other applications. The development of instrumentation capable of automated heterogeneous immunoassays (separation step to improve sensitivity) has enabled movement of this technology from the "special chemistry" sections of a clinical laboratory into the "core" laboratory with other high-volume testing. CONCLUSION: Today, immunoassays play a prominent role in the analysis of many clinical laboratory analytes such as proteins, hormones, drugs, and nucleic acids. The future involves development of assays with higher sensitivities which will enable the discovery of new biomarkers for disease diagnosis, and technology that will enable simultaneous multimarker analysis of tests whose needs are naturally grouped together (e.g., cytokines and allergens). PMID- 16701600 TI - New HDL-cholesterol reagent formulation reduces interference from samples with abnormal proteins. PMID- 16701601 TI - Multiplex PCR-pyrosequencing assay for genotyping CYP3A5 polymorphisms. AB - BACKGROUND: The cytochrome P450 (CYP) 3A5 enzyme contributes to the metabolism of many drugs. Single nucleotide polymorphisms in the CYP3A5 gene (CYP3A5(*)3C and CYP3A5(*)6) are associated with decreased CYP3A5 expression in the liver. We designed a multiplex genotyping assay to detect the CYP3A5(*)3C and CYP3A5(*)6 polymorphisms in a single polymerase chain reaction (PCR) and a single pyrosequencing reaction. METHODS: A multiplex PCR assay was designed to simultaneously amplify 2 fragments, one containing the CYP3A5(*)3C polymorphism and the other containing the CYP3A5(*)6 polymorphism. Following PCR, multiplex genotyping was performed with pyrosequencing analysis. RESULTS: Patient samples (n=69) were analyzed for the CYP3A5(*)3C and CYP3A5(*)6 polymorphisms using the multiplex PCR-pyrosequencing assay. Genotypes obtained by the multiplex reaction were in 100% concordance with genotypes obtained using simplex PCR-pyrosequencing (n=69) and direct DNA sequencing (n=29). CONCLUSIONS: The advantage of this method is that the CYP3A5(*)3C and CYP3A5(*)6 polymorphism can be amplified in a single PCR reaction and genotyped in a single pyrosequencing reaction. This combined approach improves the time-efficiency and decreases the cost of CYP3A5 genotyping. PMID- 16701603 TI - Molecular diagnostics: a historical perspective. AB - The rapid growth in molecular diagnostic testing, which has averaged between 10% and 20% per year for the past 5 years, is largely attributable to both breakthroughs in our basic understanding (i.e., the Human Genome Project) and in applied technology. In the past decade, molecular applications have moved from labor-intensive and manual to rapid and automated due to improvements in sample extraction, target amplification, and sensitive and specific detection schema. This review describes some of the more significant technological milestones of the past 10 years and, when tied to basic and applied research, how these have led to important clinical applications. The next decade promises even more exciting technologies and applications for the field of molecular laboratory medicine. PMID- 16701602 TI - Lipoprotein (a) and comprehensive lipid tetrad index as a marker for coronary artery disease in NIDDM patients in South India. AB - BACKGROUND: Coronary artery disease (CAD) is reaching epidemic proportions in India, in the absence of traditional risk factors. Lipoprotein (a) (Lp(a)) concentrations are related to both atherogenesis and thrombogenesis and may be a key link between lipid and CAD. We studied the role of Lp(a) and comprehensive lipid tetrad index as markers for CAD in South Indian patients with non-insulin dependent diabetes mellitus (NIDDM). METHODS: Lp(a) concentrations and lipid profile were estimated in 53 NIDDM patients with CAD (Group 1), 53 NIDDM patients without CAD (Group 2), and 52 control subjects (Group 3). Comprehensive lipid tetrad index was calculated in all patients and controls. RESULTS: Lp(a) concentrations were significantly higher in Group 1 patients, when compared with Groups 2 and 3. In NIDDM patients with CAD, only total cholesterol and low density cholesterol concentrations correlated significantly positively with lipoprotein (a) concentrations (r=0.184, p=0.03 and r=0.168, p=0.02). Mean comprehensive lipid tetrad index was 45,487+/-2747 in Group 1, 10,866+/-1163 in Group 2 and 4582+/-348 in Group 3 subjects. CONCLUSION: Based on the foregoing data, high Lp(a) concentrations show strong correlation with CAD in NIDDM patients of South India. High concentrations of Lp(a) and comprehensive lipid tetrad index, along with high prevalence of NIDDM, may render Indians particularly vulnerable to malignant atherosclerosis at a young age. As NIDDM is increasing in prevalence in India, the above observations have ominous dimensions in terms of total burden of CAD in India. PMID- 16701604 TI - Role of ascorbic acid on in vitro oxidation of low-density lipoprotein derived from hypercholesterolemic patients. AB - BACKGROUND: The susceptibility of low-density lipoprotein (LDL) to oxidation is thought to be a crucial factor responsible for atherogenesis. There is substantial evidence for a role of dietary antioxidants in the prevention of atherogenesis and the protective effect of antioxidant nutrients may be mediated through inhibition of the oxidative modification of LDL. METHODS: We performed in vitro oxidation of LDL derived from normal and hypercholesterolemic individuals in absence and presence of different doses of ascorbic acid. RESULTS: The serum lipid peroxidation level was significantly increased in hypercholesterolemic patients and their LDL has shown a greater propensity towards in vitro oxidation. Hypercholesterolemic LDL required a higher amount of ascorbic acid to reduce its oxidation level as compared to LDL isolated from normocholesterolemic individuals. CONCLUSION: This observation may be of importance in designing future studies of antioxidant supplementation in patients with hypercholesterolemia which is one of the major risk factors for atherosclerosis. PMID- 16701605 TI - Clostridium difficile: an important pathogen of food animals. AB - Human Clostridium difficile-associated disease (CDAD) is of unquestioned importance in humans, and has been a not-uncommon cause of enteric disease in horses, dogs, and ratites. Over the past 5 years, C. difficile has emerged as a major cause of neonatal enteritis in pigs. Piglets 1-7 days of age are affected, with gross lesions frequently including mesocolonic edema. Colonic contents may be pasty-to-watery and yellow, although some piglets are constipated or obstipated. Focal suppuration and segmental necrosis are seen on microscopic examination of cecal and colonic lamina propria, and exudation of neutrophils and fibrin into the lumen gives rise to the so-called volcano lesions. Results of one study revealed that more than one-third of piglets with enteritis were affected by C. difficile alone, while an additional quarter of affected piglets may have had mixed infections. C. difficile may be the most important uncontrolled cause of neonatal diarrhea in pigs. PMID- 16701606 TI - The role of anaerobic bacteria in sinusitis. AB - The normal oropharyngeal flora contained aerobic and anaerobic bacteria that can cause respiratory infections including sinusitis. Some of these bacteria can interfere with the growth of potential pathogens and may play a role in preventing infections. Anaerobic bacteria emerge as pathogens as the infection becomes chronic. This may be the result of the selective pressure of antimicrobial agents that enable resistant anaerobic organisms to survive, and from the development over time of conditions appropriate for anaerobic growth, which include the reduction in oxygen tension and an increase in acidity within the sinus cavity. Anaerobes were isolated in acute maxillary sinusitis of odontogenic origin and in over half of the patients with chronic sinusitis whenever proper techniques for their cultivation were employed. These organisms were also recovered in acute sinusitis that was associated with dental infections. The predominant isolates were pigmented Prevotella and Porphyromonas, Fusobacterium and Peptostreptococcus spp. PMID- 16701607 TI - Isolation and 16S ribosomal RNA gene sequence-based identification of Clostridium scindens from an intra-abdominal abscess. AB - Clostridium scindens has not been previously associated with human infection. We describe a case of an adolescent female with Crohn's disease presenting with a post-surgical intra-abdominal abscess from which this organism was isolated in pure culture. This is the first documented report of human infection caused by this micro-organism. PMID- 16701608 TI - Characterization of the Bacteroides fragilis pathogenicity island in human blood culture isolates. AB - Bacteroides fragilis is an important anaerobic pathogen accounting for up to 10% of bacteremias in adult patients. Enterotoxin producing B. fragilis (ETBF) strains have been identified as enteric pathogens of children and adults. In order to further characterize the B. fragilis pathogenicity island (BfPAI) and using PCR assays for bft- and mpII-metalloprotease genes, we determined the frequency of B. fragilis strains with pattern I (containing the BfPAI and its flanking region), pattern II (lacking both the BfPAI and the flanking region), and pattern III (lacking the BfPAI but containing the flanking region) in 63 blood culture isolates. The results were compared to 197 B. fragilis isolates from different clinical sources. We found 19% of blood culture isolates were pattern I (ETBF), 43% were pattern II (NTBF) and 38% were pattern III (NTBF). Comparatively, B. fragilis isolates from other clinical sources were 10% pattern I, 47% pattern II and 43% pattern III. This suggests that the pathogenicity island and the flanking elements may be general virulence factors of B. fragilis. PMID- 16701609 TI - Clostridium perfringens: insight into virulence evolution and population structure. AB - Clostridium perfringens is an important pathogen in veterinary and medical fields. Diseases caused by this organism are in many cases life threatening or fatal. At the same time, it is part of the ecological community of the intestinal tract of man and animals. Virulence in this species is not fully understood and it does seem that there is erratic distribution of the toxin/enzyme genes within C. perfringens population. We used the recently developed multiple-locus variable number tandem repeat analysis (MLVA) scheme to investigate the evolution of virulence and population structure of this species. Analysis of the phylogenetic signal indicates that acquisition of the major toxin genes as well as other plasmid-borne toxin genes is a recent evolutionary event and their maintenance is essentially a function of the selective advantage they confer in certain niches under different conditions. In addition, it indicates the ability of virulent strains to cause disease in different host species. More interestingly, there is evidence that certain normal flora strains are virulent when they gain access to a different host species. Analysis of the population structure indicates that recombination events are the major tool that shapes the population and this panmixia is interrupted by frequent clonal expansion that mostly corresponds to disease processes. The signature of positive selection was detected in alpha toxin gene, suggesting the possibility of adaptive alleles on the other chromosomally encoded determinants. Finally, C. perfringens proved to have a dynamic population and availability of more genome sequences and use of comparative proteomics and animal modeling would provide more insight into the virulence of this organism. PMID- 16701610 TI - Immunization with an alphatoxin variant 121A/91-R212H protects mice against Clostridium perfringens alphatoxin. AB - As shown previously, a recombinant alphatoxin variant (rAT121A/91) constructed from the naturally occurring Clostridium perfringens mutant strain 121A/91, was devoid of enzymatic (PLC), hemolytic and lethal activity (18). In the present study, the recombinant variant was altered by an oligonucleotide-directed reversion of an arginine in position 212 for a histidine residue, corresponding to the sequence of the wild-type alphatoxin. The new variant rAT121A/91R212H proved to be negative in enzymatic, hemolytic and lethal activity as well. RAT121A/91 as well as rAT121A/91R212H was used for i.p. immunization of balb/c mice. The immune response was studied in ELISA as well as in the mouse neutralization test. Furthermore, immunized mice were challenged by i.p. application of active C. perfringens alphatoxin. In all immunized groups, mice developed high anti-alphatoxin titers (up to 1:128000). Antisera of both groups were able to reduce the hemolytic effect of native alphatoxin with predominance of anti-rAT121A/91R212H sera. During neutralization experiments, mice receiving a mixture of anti-rAT121A/91R212H and wild-type toxin were protected completely, whereas an anti-rAT121A/91/toxin mixture prolonged time until death but failed in protection. I.p immunization with rAT121A/91R212H yielded a significant protection rate (76%) when mice were challenged intraperitoneal with wild-type toxin. Our cumulative data indicates that the reversion of arginine in position 212 to histidine for rAT121A/91R212H was necessary to induce production of protective antibodies against wild-type alphatoxin of C. perfringens. PMID- 16701611 TI - Convenient anaerobic techniques, science from the supermarket shelf. AB - We describe the application and evaluation of a widely available commercial jar as an anaerobic container suitable for the growth of a wide variety of anaerobes. A system for generating stable anaerobiosis was developed by combining standard anaerobic environment generators with Click-Clack jars produced by Click-Clack Ltd. This system was simple, reliable, and reduced capital outlay on anaerobic jars by at least an order of magnitude. PMID- 16701612 TI - Distribution of anaerobic bacteria among pregnant periodontitis patients who experience preterm delivery. AB - The preterm delivery of low-birth-weight (PLBW) infants remains a significant public health issue and a leading cause of neonatal death and of long-term neurodevelopmental disturbances and health problems. Epidemiological and immunological studies have suggested that periodontal disease may be an independent risk factor for PLBW. The incidence of periodontal disease during pregnancy has been found to be associated with a significantly lower birth weight for gestational age after adjustment for race, parity and baby gender. The present case-control study, carried out to examine the distribution of anaerobes in pregnant women with periodontitis who experienced preterm delivery, provides microbial evidence that maternal periodontal disease and the presence of key pathogens are significant contributors to the obstetric risk of preterm delivery. PMID- 16701613 TI - Generalized training subset selection for statistical estimation of epicardial activation maps from intravenous catheter measurements. AB - Catheter-based electrophysiological studies of the epicardium are limited to regions near the coronary vessels or require transthoracic access. We have developed a statistical approach by which to estimate high-resolution maps of epicardial activation from very low-resolution multi-electrode venous catheter measurements. This technique uses a linear estimation model that derives a relationship between venous catheter measurements and unmeasured epicardial sites from a set of previously recorded, high-resolution epicardial activation-time maps used as a training data set based on the spatial covariance of the measurement sites. We performed 14 dog experiments with various interventions to create an epicardial activation-time map database. This database included a total of 592 epicardial activation maps which were recorded using a sock array placed on the ventricles of dog hearts. We present five approaches, which examined sequential addition and removal of maps to select a generalized training set for the estimation technique. The selection consisted of choosing a subset of epicardial ectopic activation-time maps from the database of beats which resulted in estimation accuracy levels better than or at least similar to using all the maps in database. Our aim was to minimize the redundancy in the database and to be able to guide the eventual procedures required to obtain training data from open-chest surgery patients. The results from this study illustrated this redundancy and suggested that by including an optimal subset (around 100 maps) of the full database the estimation technique was able to perform as well as and even in some cases better than including all the maps in the database. The results also suggest that such an approach is feasible for providing accurate reconstruction of complete epicardial activation-time maps in a clinical setting and with fewer maps we can obtain similar reconstruction accuracy levels. PMID- 16701614 TI - Health protection: microbiology and the public health response to the bioterrorism threat. PMID- 16701615 TI - In vitro activity of 11 antibiotics against 74 anaerobes isolated from pediatric intra-abdominal infections. AB - The in vitro activity of 11 antimicrobials was tested against 74 recent anaerobic isolates obtained from pretreatment cultures in pediatric patients with complicated intra-abdominal infections using the CLSI M11-A-6 agar dilution method. Carbapenems, beta-lactamase inhibitor combinations and metronidazole retained good activity, while all Bacteroides fragilis group species produced beta-lactamase and were penicillin resistant and 43% were either intermediately susceptible or resistant to clindamycin. Cefoxitin had moderate activity against B. fragilis but poor activity against Bacteroides thetaiotaomicron and other B. fragilis group isolates. PMID- 16701616 TI - Intrauterine contraceptive device-associated pelvic actinomycosis caused by Actinomyces urogenitalis. AB - We report a case of intrauterine contraceptive device (IUD) associated pelvic actinomycosis due to Actinomyces urogenitalis in a previously healthy young adult woman. Diagnosis was confirmed by 16S ribosomal RNA gene sequencing of bacterial colonies growing from the extracted device. This is the first documented report of human infection caused by this micro-organism. PMID- 16701617 TI - The distribution of the bft alleles among enterotoxigenic Bacteroides fragilis strains from stool specimens and extraintestinal sites. AB - Enterotoxigenic Bacteroides fragilis (ETBF) has been implicated in diarrhoeal illness in animals and humans. Recent data suggest that ETBF is associated with flares of inflammatory bowel disease. Toxigenicity is attributed to expression of a toxin referred to as fragilysin, which stimulates fluid accumulation in ligated intestinal segments and alter the morphology of human intestinal cells. Three different isoforms or variants of the enterotoxin gene, designated bft-1, bft-2, and bft-3, have been identified. In this study we investigated the distribution of bft alleles among ETBF strains in stool specimens from patients with colon cancer (n: 31), the control patients (n: 8) and extraintestinal sources (n: 15). We used restriction fragment length polymorphism analysis of the PCR-amplified enterotoxin gene and sequencing the PCR-product to detect the isoforms of bft gene. Among the stool strains, bft-1 was found to be more common than bft-2; as it was detected 27 of 31 strains from colon cancer patients and 7 of 8 control strains. The bft-1 isoform was also found in almost all isolates from extraintestinal sites. No bft-3 subtype was detected among all tested strains. PMID- 16701618 TI - Application of potential probiotic Lactobacillus fermentum AD1 strain in healthy dogs. AB - Probiotic utilization is becoming increasingly popular in veterinary medicine. However, only few probiotic products are available commercially for use in dogs in our market. Therefore, the aim of our study was to determine the properties of new potential probiotic Lactobacillus fermentum AD1 strain-own canine isolate and to investigate its effect on several microbiological and biochemical parameters in healthy dogs. The strain expressed in vitro survival by pH 3.0 after 3h (86.8%) and in the presence of 1% bile (75.4%). The AD1 strain adhered to the canine and human intestinal mucus. It was sensitive to commonly used antimicrobials. Fifteen healthy dogs were supplemented with 10(9)L. fermentum AD1 for 7 days. At the end of AD1 strain application, numbers of faecal lactobacilli and enterococci increased significantly in the canine faeces. Significant increase of total protein and total lipid and significant reduction of glucose in serum of dogs were noted. These data indicate that L. fermentum AD1 survive transit through the canine gastrointestinal tract, and populate the colon and probably increased absorption of some nutrients. Whether longer time of its application lead to the same results as well as its potential to improve immune function in dogs remains to be determined. PMID- 16701619 TI - Influence of phenol, p-cresol and indole on growth and survival of intestinal lactic acid bacteria. AB - Some intestinal bacteria can produce many genotoxic, mutagenic and carcinogenic substances. The major products of the bacterial aromatic amino acids fermentation phenolic and indolic compounds which are responsible for colon cancer development are accumulated in the colon. The effect of phenol, p-cresol and indole (2, 20 and 100 microg/ml doses) on growth and survival of four strains of intestinal lactic acid bacteria was studied. Growth of bacteria was not affected by any of the concentrations of phenol and p-cresol tested. The growth of 2 strains was slightly inhibited by 100 microg/ml of indole. There was no influence of phenol and p-cresol on survival of lactic bacteria until 120 h and specific reaction to carcinogens depending on strain was observed after that incubation time. Indole concentrations 20 and 100 microg/ml appeared to be toxic for all tested strains but just after 24, 48 or 72 h of incubation depending on the strain. In total, 2 microg/ml of indole had a very little effect. PMID- 16701620 TI - Effects of nitro compounds and feedstuffs on in vitro methane production in chicken cecal contents and rumen fluid. AB - Short-chain volatile fatty acids (VFA) and methane are the products from a wide variety of microorganisms living in the gastrointestinal tract. The objective of this study was to examine effects of feedstuff and select nitro compounds on VFA and methane production during in vitro incubation of laying hen cecal contents and rumen fluid from cattle and sheep. In the first experiment, one of the three nitro compound was added to incubations containing cecal contents from laying hens supplemented with either alfalfa (AF) or layer feed (LF). Both feed material influenced VFA production and acetic acid was the primary component. Incubations with nitro ethanol and 2-nitropropanol (NP) had significantly (P<0.05) higher propionate concentrations than incubations with added nitroethane (NE). The results further indicated that incubations containing LF produced significantly (P<0.05) more butyrate than incubations with added AF. Addition of NP and LF to incubations of avian cecal flora may promote Gram-positive, saccharolytic, VFA producing bacteria, especially Clostridium spp. which is the predominant group in ceca. Similar to VFA production, both feed materials fostered methane production in the incubations although methane was lower (P<0.05) in incubations with added nitro compound, particularly NE. In experiments 3-8, NE was examined in incubations of bovine or ovine rumen fluid or cecal contents containing either AF or LF. Unlike cecal contents, LF significantly (P<0.05) supported in vitro methane production in incubations of both rumen fluids. The results show that NE impedes methane production, especially in incubations of chicken cecal contents. PMID- 16701621 TI - Evaluation of support materials for the immobilization of sulfate-reducing bacteria and methanogenic archaea. AB - This paper reports on the adhesion of sulfate-reducing bacteria (SRB) and methanogenic archaea on polyurethane foam (PU), vegetal carbon (VC), low-density polyethylene (PE) and alumina-based ceramics (CE). Anaerobic differential reactors fed with a sulfate-rich synthetic wastewater were used to evaluate the formation of a biofilm. The PU presented the highest specific biomass concentration throughout the experiment, achieving 872 mg TVS/g support, while 84 mg TVS/g support was the maximum value obtained for the other materials. FISH results showed that bacterial cells rather than archaeal cells were predominant on the biofilms. These cells, detected with EUB338 probe, accounted for 76.2% (+/ 1.6%), 79.7% (+/-1.3%), 84.4% (+/-1.4%) and 60.2% (+/-1.0%) in PU, VC, PE and CE, respectively, of the 4'6-diamidino-2-phenylindole (DAPI)-stained cells. From these percentages, 44.8% (+/-2.1%), 55.4% (+/-1.2%), 32.7% (+/-1.4%) and 18.1% (+/-1.1%), respectively, represented the SRB group. Archaeal cells, detected with ARC915 probe, accounted for 33.1% (+/-1.6%), 25.4% (+/-1.3%), 22.6% (+/-1.1%) and 41.9% (+/-1.0%) in PU, VC, PE and CE, respectively, of the DAPI-stained cells. Sulfate reduction efficiencies of 39% and 45% and mean chemical oxygen demand (COD) removal efficiencies of 86% and 90% were achieved for PU and VC, respectively. The other two supports, PE and CE, provided mean COD removal efficiencies of 84% and 86%, respectively. However, no sulfate reduction was observed with these supports. PMID- 16701622 TI - The effect of traditional African food mixtures on growth, pH and extracellular polysaccharide production by mutans streptococci in vitro. AB - Four, traditional African food mixtures (maize plus milk and sugar, maize plus gravy, samp plus beans, brown bread plus margarine and peanut butter) were evaluated for their ability to sustain the growth of mutans streptococci in batch culture. A synthetic complex medium, brain heart infusion with 3% sucrose was used as an experimental control. Six NCTC laboratory reference strains and five clinical isolates collected from the plaque of children were investigated. The doubling time of bacterial strains was prolonged in maize plus gravy (2.5-6.0 h) and samp plus beans (1.3-9.9h), and the number of cell divisions was low, compared with bread plus margarine plus peanut butter (0.7-5.1h). The least amount of acid was produced in maize plus milk plus sugar (3.92+/-8.15 mmole/mL), and the average pH during the fermentation of maize plus milk plus sugar, maize plus gravy and samp plus beans did not drop below the critical point for enamel demineralisation, pH 5.7. Bacterial growth in samp plus beans produced a small quantity of lactic acid (0.46+/-1.10 mmole/mL) compared to bread plus margarine and peanut butter (2.64+/-3.30 mmole/mL) and BHI plus 3% sucrose (12.23+/-10.72 mmole/mL). Extracellular polysaccharide (ECP) produced was lowest in maize plus milk and sugar (0.22+/-0.33 mg/mL), compared with the remaining food mixtures (0.47-1.75 mg/mL). Statistical analysis showed that the influence of the mixed foods on doubling time (F=3.01, P=0.03), pH (F=14.41, P<0.0001) and ECP (F=135.32, P<0.0001) was greater than the significant variance found between mutans streptococci strains. Results suggest that the level of mutans streptococci activity in samp plus beans, maize plus milk and sugar and maize plus gravy contributes little towards the formation of dental caries, and that significant differences exist between mutans streptococci laboratory reference and clinical strains in response to traditional African food mixtures. PMID- 16701623 TI - AFM study of the effect of metronidazole on surface structures of sulfate reducing bacteria. AB - The effect of metronidazole (ME) on sulfate-reducing bacteria (SRB) was studied by atomic force microscopy (AFM) in this paper. Topography images of SRB cell show that after exposure to ME individual cell shape is sharply modified. Topography and phase images of SRB cell wall show that after exposure to ME not only the roughness of the cell wall increases but also the physical performance of SRB surface is changed to be uniform. AFM frictional loops show that after exposure to ME, SRB surface friction is increased remarkably. PMID- 16701624 TI - D-ribose utilisation differentiates porcine Brachyspira pilosicoli from other porcine Brachyspira species. AB - D-ribose utilisation was studied in 60 Brachyspira pilosicoli strains and 35 strains of other Brachyspira species, the majority of which were of porcine origin. Utilisation of D-ribose was demonstrated indirectly by measuring the reduction in pH of densely inoculated tryptone-peptone broth supplemented with 7% foetal calf serum and 1% D-ribose. Among B. pilosicoli strains, the mean reduction in pH units was 1.72 (range 0.95-2.28) in broth with D-ribose and 0.27 (range 0.10-0.40) in sugar-free control broth. For Brachyspira strains other than B. pilosicoli, the corresponding reductions in pH units were 0.37 (range 0.12 0.49) and 0.37 (range 0.15-0.58). In conclusion, porcine B. pilosicoli can be differentiated from other porcine Brachyspira species by a test for D-ribose utilisation. PMID- 16701627 TI - FEV1 or peak flow for measuring airflow obstruction in primary care. Is it a useful debate or just missing the point? PMID- 16701626 TI - What's in this issue. PMID- 16701625 TI - The T-box factor TBX-2 and the SUMO conjugating enzyme UBC-9 are required for ABa derived pharyngeal muscle in C. elegans. AB - The C. elegans pharynx is produced from the embryonic blastomeres ABa and MS. Pharyngeal fate in the ABa lineage is specified by the combined activities of GLP 1/Notch-mediated signals and the TBX-37 and TBX-38 T-box transcription factors. Here, we show another T-box factor TBX-2 also functions in ABa-derived pharyngeal development. tbx-2 mutants arrest as L1 larvae lacking most or all ABa-derived pharyngeal muscles. In comparison, tbx-2 mutants retain ABa-derived marginal cells and pharyngeal muscles derived from MS. A tbx-2Colon, two colonsgfp translational fusion is expressed in a dynamic pattern in C. elegans embryos beginning near the 100-cell stage. Early expression is limited to a small number of cells, which likely include the ABa-derived pharyngeal precursors, while later expression is observed in body wall muscles and a subset of pharyngeal neurons. TBX-2 contains 2 consensus sumoylation sites, and it interacts in a yeast two hybrid assay with the UBC-9 and GEI-17 components of the C. elegans SUMO conjugating pathway. ubc-9(RNAi) has been previously shown to cause variable embryonic and larval arrest, and we find that, like tbx-2 mutants, ubc-9(RNAi) animals lack ABa-derived pharyngeal muscles. ubc-9(RNAi) also alters the subnuclear distribution of TBX-2::GFP fusion protein, suggesting that UBC-9 and TBX-2 interact in C. elegans. Together, these results indicate that TBX-2 and SUMO-conjugating enzymes are necessary for ABa-derived pharyngeal muscle, and we hypothesize that TBX-2 function requires sumoylation. Sumoylation is increasingly recognized as an important mechanism controlling activity of many nuclear factors, and these results provide the first evidence that T-box factor activity may require sumoylation. PMID- 16701628 TI - Spirometry and peak expiratory flow in the primary care management of COPD. AB - Spirometry is essential for the diagnosis of chronic obstructive pulmonary disease (COPD). In patients with COPD the decline in lung function is usually so slow that spirometry is unlikely to provide significantly new information more than every 1-2 years. However, it is useful to have an objective measure of lung function in the assessment of acute exacerbations of COPD and in the assessment of treatments. Peak expiratory flow (PEF) has been dismissed by national and international guidelines as an inappropriate test for the assessment of the impact of COPD, but with poor evidence in support of this position. This seems short-sighted since PEF is a reliable and reproducible test and could contribute to the management of COPD in the short term and in support of spirometry. As a result of infection or in response to treatment there may be changes in airway calibre in COPD which could be captured in the consultation by PEF. In a primary care setting spirometry is too time consuming and complex to be provided in the context of normal acute consulting. Furthermore there is no evidence that spirometry provides more information than PEF in the day-to-day management of a patient already diagnosed with COPD using forced expiratory volume in the first second/forced vital capacity (FEV1/FVC). Primary care teams should ensure that their patients have adequate access to high quality spirometry. This can be provided in primary care or in local centres or in hospitals depending on the interest, motivation and resources of primary care teams. In support of spirometry general practitioners (GPs) should then consider using PEF in the day to-day management of COPD. PMID- 16701629 TI - Reply to: Spirometry and peak expiratory flow in the primary care management of COPD by Patrick White. PMID- 16701630 TI - The necessity for spirometry in the primary care management of COPD. AB - Implementation of spirometry in the primary care setting is controversial. Spirometry allows a unique non-invasive look into the functioning of the lungs, which can be both medically informative and of practical value. Respiratory complaints are among the most prevalent in primary care, while smoking rates remain globally high, illustrating the need for proper tools to investigate the possible causes of chronic pulmonary symptoms. Smoking cessation programs and disease management programs are the preferred treatment strategies for COPD; these rely on a valid spirometric diagnosis, as promoted by international COPD guidelines. In addition, involving the patient in treatment plans and explaining the detrimental effects of smoking can be greatly facilitated by the visual impact of a flow-volume curve, have promoted the use of spirometry. While there are difficulties in implementation of spirometry in primary care. Provided that quality standards of performance and training are sufficiently met, evidence suggests that spirometry is a valid, feasible and interpretable diagnostic tool in a primary care setting. COPD is a growing problem for society, which should not be underestimated, and with sufficient funding, adequate training and motivated healthcare workers, there is no good reason why spirometric testing cannot be widely implemented. PMID- 16701631 TI - Response to: The necessity for spirometry in the primary care management of COPD. PMID- 16701632 TI - Asthma action plans: use it or lose it. PMID- 16701633 TI - The use of the self-management plan system of care in adult asthma. AB - The development of self-management plans arose as clinicians tried to design better methods by which they could deliver asthma care and reduce the significant mortality and morbidity associated with this disease. The basic principles that resulted have been widely endorsed, and self-management plans are now recommended in the long-term management of adult asthma. Self-management plans essentially focus on the early recognition of unstable or deteriorating asthma, by monitoring peak flow or symptoms. Through the use of written guidelines, patients are then able to determine when it is necessary to adjust therapy or obtain medical assistance. There is now convincing evidence that the use of self-management plans by patients with asthma leads to a marked reduction in morbidity and a reduced requirement for acute medical treatment including hospital admissions. Recent research has also clarified many of the different issues concerning their structure and implementation. In some respects the skill in the use of the asthma self-management plan system of care is the ability to modify the standard plans to meet the particular needs of the individual asthmatic patient, including their preferences. PMID- 16701634 TI - The Living & Breathing Study: a study of patients' views of asthma and its treatment. AB - AIM: To evaluate patient understanding of their asthma and determine patient preferences regarding the delivery of asthma care and treatment. METHODS: Adults with asthma receiving treatment for mild to moderate asthma were recruited to a two-part study: a qualitative phase using a semi-structured interview schedule followed by a quantitative phase based on a structured interview schedule. All interviews were undertaken face-to-face. SETTING AND SUBJECTS: A random sample of 40 patients with mild to moderate asthma from seven areas of the UK took part in the qualitative phase of the study. In the quantitative phase, 517 patients on treatment for mild to moderate asthma were interviewed in person by market researchers. This population was achieved using a quota sampling approach that also achieved a representative demographic profile. Initial contact was made in door-to-door calls. Interviews took place in 64 locations across the UK. RESULTS: Ninety-one percent (n=468) of respondents felt their asthma was under control, yet two-thirds (n=339) experienced symptoms at least 2-3 times a week. Only 24% (n=123) felt their asthma could improve over time, and 71% (n=366) received no advice from healthcare professionals on how their asthma might change in the future. Fourteen percent (n=74) of respondents had no ongoing contact with any healthcare professional regarding their asthma. Fifty-eight percent (n=301) were very satisfied with their asthma care, but this dropped to 33% (n=173) when respondents were shown asthma guidelines regarding what to expect from treatment. Sixty-two percent (n=318) of respondents said their asthma varied at different times of the day, and 86% (n=444) stated that their asthma varied at different times of the year. Eighty percent (n=414) of respondents had never been provided with a written, personal asthma action plan recommending changes patients could make themselves to prescribed treatment according to symptom severity, though 68% (n=353) said they would feel comfortable following such a plan. CONCLUSIONS: Most patients have low expectations of what can be achieved by asthma management and do not realise their condition can be improved. Many are resigned to the effects of poor asthma control until made aware that guidelines indicate this can be better. Given that many are receptive to the notion of written, personal asthma action plans, the implementation of these, supported by appropriate education, could help patients achieve improved asthma control. PMID- 16701635 TI - Evaluation of an intervention to improve successful completion of the Mini-AQLQ: comparison of postal and supervised completion. AB - INTRODUCTION: Previous postal administration of Mini Asthma-related Quality of Life Questionnaire (Mini-AQLQ) (validated for self-completion under supervision) resulted in 12.7% completion error rate. AIMS: To administer the Mini-AQLQ by post with instructions, and to compare completion errors with our previous study and usable response rate with supervised self-completion. METHOD: The Mini-AQLQ, with an instruction sheet, was posted to 96 participants from UK general practice, for completion 1 week before supervised self-completion in the surgery. RESULTS: 94/96 (98%) postal questionnaires were returned: the error rate of 10.6% was similar to our previous study (postal versus previous: 10/94 versus 23/181: P=0.62). 86/96 (90%) attended for supervised completion with no completion errors (supervised versus postal: 0/86 versus 10/94: P0.3). Results indicate that the electrochemical behavior of the Ti-Hf alloys examined resembles that of pure titanium. PMID- 16701814 TI - Rheological characterization of concentrated aqueous beta-tricalcium phosphate suspensions: the effect of liquid-to-powder ratio, milling time, and additives. AB - The field of injectable calcium phosphate suspensions and cements is experiencing vigorous research activity. This is stimulated by their importance for the cement augmentation procedure (vertebroplasty), which is an emerging procedure to treat osteoporotic fragility fractures. The rheological properties such as the yield stress and viscosity play an important role in the process of cement delivery and infiltration into the cancellous bone cavities. However, the number of studies relating to their rheological properties is very limited. The objective of this first study was to examine the effects of the following three variables on the rheological properties of a non-setting beta-tricalcium phosphate suspension: liquid-to-powder ratio, milling of powder particles, and additives. The broad finding is that all the variables affect the rheological properties remarkably. The more specific salient finding is the large variation in viscosity and in the yield stress. The viscosity spanned three orders of magnitude and the yield stress spanned five orders of magnitude. It appears that the rheological properties can be altered at will. However, one has to exercise extreme caution because these changes are not without cost to other important properties such as the cohesiveness and mechanical properties of the cement. Another important finding is that a linear correlation between the yield stress and the viscosity was found. Measurement of one of these variables might be enough to determine the other. PMID- 16701815 TI - Preferences for sources of information about abnormal Pap tests and HPV in women tested for HPV. AB - BACKGROUND: Genital human papillomavirus (HPV) infection is important because of its association with cervical cancer. There has been a recent upsurge of coverage in the media concerning HPV, but little research exists on women's preferences and opinions regarding sources of information and education on HPV and abnormal Pap tests. The purpose of this paper is to describe women's preferences and opinions about various sources/channels of information on abnormal Pap test results and HPV. METHODS: Forty-four in-depth, in-person interviews were conducted with low-income, high-risk HPV positive women. Women responded to open ended questions addressing their preferences for sources of health and medical information related to HPV and abnormal Pap tests. The audiotaped interviews were transcribed verbatim. The transcripts were analyzed for content themes related to provision of information and education. RESULTS: Women preferred sources that were trustworthy, accessible, and convenient, could be viewed in a private location, and were written in easily understood language. Women also preferred obtaining information from other women with HPV. The most trusted source was face to-face interaction with the health care provider; however, the nature of this interaction had both strengths and weaknesses. CONCLUSIONS: Regardless of channel, women want information that they can understand about abnormal Pap tests and HPV. PMID- 16701816 TI - Early development of reconstructed embryos after somatic cell nuclear transfer in a non-human primate. AB - To improve efficiency and assess variation in nuclear transfer techniques in non human primates, we investigated the following factors: type of donor cell, interval between enucleation and cell injection, activation after electrical pulsing and cytokinesis inhibitors. An average of 16.4 oocytes were recovered from 91 retrievals; however, 15 (14%) additional retrieval attempts yielded no oocytes due to a failure of follicular stimulation. Oocyte maturation rates at 36, 38 and 40 h post-hCG were 46.2, 52.6 and 61.2%, respectively. The MII spindle could be seen clearly using polarized microscopy in 89.1% (614/689) of oocytes. Nuclei were seen in 42% of the NT couplets, 53% of those cleaved to the 2-cell stage and 63% of the 2-cell embryos developed to the 8-cell stage by Day 3. There was no difference in the occurrence of nuclear formation between couplets created using fibroblasts or cumulus cells, although embryos were more reliably produced with fibroblasts. The interval (2, 3 and 4 h) between enucleation and cell injection did not affect NT efficiency. Ethanol treatment after electrical pulses yielded more 2-cell NT embryos than did treatment with ionomycin, but the frequency of nuclear formation and development to the 8-cell stage was not different. Treatment of couplets with cycloheximide and cytochalasin B for 5 h after activation had no impact on NT efficiency. PMID- 16701817 TI - Reproductive and sexual behavioral problems in dogs. AB - This paper reviews the influence of genetics on canine sexual behavior, the causes of common sexual and reproductive behavioral problems in dogs as well as treatment approaches, including environmental management, behavior modification, and pharmacological intervention. PMID- 16701818 TI - Adhesion enhancement in a biomimetic fibrillar interface. AB - Two important putative functions of the fibrillar contact interfaces commonly found in lizards and insects are to provide contact compliance and enhanced adhesion. To explore the question of whether a fibrillar architecture inherently enhances adhesion, we constructed model structures consisting of thin sheets of poly(vinyl butyral) (PVB) bonded on one of their thin sides to glass plates. The PVB samples had two flat, unstructured regions interrupted by a central fibrillar region along the bonded interface. The effect of the fibrillar geometry on the performance of the adhesive bond was tested using a tensile pull-off test, in which failure occurred by interfacial crack propagation, starting at an end where a crack initiator was introduced. We observed that fibrils in all samples fail consistently at the same critical stress, which is consistent with a previous theoretical result we have determined for flaw insensitivity during fibrillar pull-off. In addition, we measured the energy release rate required to fail the interface in the fibrillar region to be about an order of magnitude greater than that in the non-fibrillar region. We present experimental evidence demonstrating that this increase results partly from dissipation of strain energy stored in the fibrils. PMID- 16701819 TI - Regulation of cellular infiltration into tissue engineering scaffolds composed of submicron diameter fibrils produced by electrospinning. AB - We characterize the infiltration of interstitial cells into tissue engineering scaffolds prepared with electrospun collagen, electrospun gelatin, electrospun poly(glycolic) acid (PGA), electrospun poly(lactic) acid (PLA), and an electrospun PGA/PLA co-polymer. Electrospinning conditions were optimized to produce non-woven tissue engineering scaffolds composed of individual fibrils less than 1000 nm in diameter. Each of these materials was then electrospun into a cylindrical construct with a 2 mm inside diameter with a wall thickness of 200 250 microm. Electrospun scaffolds of collagen were rapidly, and densely, infiltrated by interstitial and endothelial cells when implanted into the interstitial space of the rat vastus lateralis muscle. Functional blood vessels were evident within 7 days. In contrast, implants composed of electrospun gelatin or the bio-resorbable synthetic polymers were not infiltrated to any great extent and induced fibrosis. Our data suggests that topographical features, unique to the electrospun collagen fibril, promote cell migration and capillary formation. PMID- 16701820 TI - Self-assembling peptide amphiphile nanofiber matrices for cell entrapment. AB - We have developed a class of peptide amphiphile (PA) molecules that self-assemble into three-dimensional nanofiber networks under physiological conditions in the presence of polyvalent metal ions. The assembly can be triggered by adding PA solutions to cell culture media or other synthetic physiological fluids containing polyvalent metal ions. When the fluids contain suspended cells, PA self-assembly entraps cells in the nanofibrillar matrix, and the cells survive in culture for at least three weeks. We also show that entrapment does not arrest cell proliferation and motility. Biochemical and ultrastructural analysis by electron microscopy indicate that entrapped cells internalize the nanofibers and possibly utilize PA molecules in their metabolic pathways. These results demonstrate that PA nanofibrillar matrices have the potential to be used for cell transplantation or other tissue engineering applications. PMID- 16701821 TI - Three-dimensional co-culture of rat hepatocyte spheroids and NIH/3T3 fibroblasts enhances hepatocyte functional maintenance. AB - Functional maintenance of primary hepatocytes in culture can be improved by several distinct approaches involving optimization of the extracellular matrix microenvironment, media composition and cell-cell interactions, both homotypic and heterotypic. Using a galactose-decorated surface, we have developed a method to combine these two approaches by co-culturing rat primary hepatocyte spheroids with NIH/3T3 mouse fibroblast cells. Spheroids were performed by culturing hepatocytes for 3 days on galactosylated poly(vinylidene difluoride) membrane; NIH/3T3 cells were subsequently seeded and co-cultured with the spheroids. Results showed that although NIH/3T3 cells alone responded poorly to the galactosylated PVDF surface and displayed limited attachment, NIH/3T3 fibroblasts attached to the periphery of the hepatocyte spheroids and proliferated around them. Co-cultured hepatocyte spheroids exhibited significantly higher liver specific functions as compared to spheroids cultured alone. Albumin secretion level in this co-culture system peaked on day 11, which was 1.8- and 2.9-times higher than the peak expression level in spheroid homo-culture control in serum free (day 3) and serum-containing media (day 4), respectively. The albumin secretion function was maintained for at least two weeks; it was 5.1 (in serum free medium) and 17.8 (in serum-containing medium) times higher than spheroid homo-culture on day 13. Similarly, the co-culture system also expressed approximately 5.5- and 3.1-times higher 3-methylcholanthrene-induced cytochrome P450 enzymatic activity on day 14 as compared to the homo-culture control in serum-free and serum-containing medium, respectively. In conclusion, this unique co-culture system demonstrated the synergistic roles of homotypic cell-cell interaction, heterotypic cell-cell interaction, cell-substrate interaction and soluble stimuli in hepatocyte functional maintenance. PMID- 16701822 TI - In vitro degradation behavior of a novel bioresorbable composite material based on PLA and a soluble CaP glass. AB - Poly(alpha-hydroxy acids), and in particular polylactic acid (PLA), are nowadays amongst the most used bioabsorbable materials. However, this polymer may not meet some application requirements due to inadequate mechanical properties and or its degradation characteristics. A possible strategy to tackle this problem is the incorporation of an inorganic phase into the polymeric matrix. In this work a new fully biodegradable composite material made with PLA and calcium phosphate soluble glass particles has been developed. The behaviour of the PLA/glass composite has been analysed during its degradation in simulated physiological conditions by means of weight loss, molecular weight and thermal properties analysis and electron microscopy observation. The results showed that the incorporation of phosphate glass particles into the polymer significantly accelerated the degradation of the PLA and induced the formation of calcium phosphate precipitates at the composite surface. PMID- 16701823 TI - The influence of crystallised Fe3O4 on the magnetic properties of coprecipitation derived ferrimagnetic glass-ceramics. AB - Ferrimagnetic glass-ceramics are potential candidates for magnetic induction hyperthermia, which is one form of inducing deep-regional hyperthermia, by using a magnetic field. The aim of this work was to analyse the influence of the amount of crystallised magnetite on the magnetic properties of glass-ceramic samples. Thus, two different ferrimagnetic glass-ceramics with the composition of the system Na(2)O-CaO-SiO(2)-P(2)O(5)-FeO-Fe(2)O(3) were prepared by melting at 1500 degrees C for 30 min of the coprecipitation-derived starting products. The X-ray diffraction patterns show the presence of nanometric magnetite crystals in a glassy matrix after cooling from melting temperature. The estimated amount of crystallised magnetite varies between 20 and 45 wt.%, as a function of the chemical composition. The morphology of the crystals was studied by scanning electron micrography and transmission electron micrography. Glass transition temperature and thermal stability were investigated by differential thermal analysis. Magnetic hysteresis cycles were analysed using a vibrating sample magnetometer with a maximum applied field of 17 kOe, at room temperature, in quasi-static conditions. Calorimetric measurements were carried out using a magnetic induction furnace. The power losses estimated from calorimetric measurements under a magnetic field of 40 kA/m and 440 kHz are 65 W/g for the glass-ceramic with lower iron oxides content and 25 W/g for the glass-ceramic with higher iron oxide content. PMID- 16701824 TI - Tailoring surface properties of biomedical polymers by implantation of Ar and He ions. AB - Ion implantation at 25 and 100 keV has been used as a tool for the modification of the surface properties of two biomedical polymers. The modulation induced by the different energy dispersion mechanisms of Ar and He have allowed satisfactory modifications for both the activation of the surfaces of chemically functional polycaprolactone (PCL) and the stabilization of anti-fouling poly(ethylene glycol) (PEG). In both cases the implantations have been performed at doses of 10(14) cm(-2) by taking into account the effect of different current densities, which are shown to distinctly influence the fragmentation-crosslinking of the target polymers. The resultant films were characterized by Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, time of flight secondary ion mass spectroscopy and atomic force microscopy. Both shifts in zeta potential versus pH curves and the alteration of the polar components of the surface free energy (contact angle measurements) were correlated with the composition analysis. The response of the modified surfaces towards biomolecular interaction is demonstrated by the induction of preferential adsorption on irradiated PCL and the inhibited adsorption onto implanted PEG regions for selected oligopeptides and proteins. PMID- 16701825 TI - Immobilization of an anticoagulant benzamidine derivative: effect of spacer arms and carrier hydrophobicity on thrombin binding. AB - Prevention of blood coagulation is very often a prerequisite for successful medical devices. For that purpose, passivation of the key coagulation enzyme thrombin through the derivatization of the material's surface with an amidine based molecule has been found to be promising. To further enhance the efficiency of this approach, thin layers of maleic anhydride copolymers offering different physico-chemical characteristics were tethered with carboxyl terminated polyethylene glycol to covalently immobilize a benzamidine-type derivative. The free carboxyl surface groups produced by the attachment of polyethylene glycol (PEG) were quantified by Ag(+) labeling and subsequent XPS detection. The film thickness as well as the carboxyl group content were found to be clearly dependent on the copolymer hydrophobicity and the nature of the PEG molecule. For the assessment of the anchorage of the thrombin to the benzamidine-derivative functionalized surfaces, the substrates were immersed in a buffered thrombin solution and the enzyme adsorption was studied using immunostaining/confocal laser scanning microscopy. Higher degrees of thrombin binding were observed for substrates configured with the hydrophilic compared to the more hydrophobic copolymer. Moreover, surface-bound spacers based on alpha,omega heterobifunctional PEG amino acids (alphaAm,omegaAc-PEG) also enhanced the benzamidine surface density in comparison to homofunctional PEG diacids (alphaAc,omegaAc-PEG) because of a lower degree of carboxyl inactivation due to PEG 'bridging'. Altogether, the choice of copolymer coatings and the type of PEG spacers were demonstrated to enhance the efficiency of the thrombin scavenging by the covalently immobilized coagulation inhibitor. PMID- 16701826 TI - Discovery of a sulfated tetrapeptide that binds to vascular endothelial growth factor. AB - Molecules that mimic the sulfated glycosaminoglycan heparin and bind to heparin binding growth factors would serve as important building blocks for synthetic biomaterials, e.g. to create a growth factor reservoir within a matrix. Peptide based heparin mimetics would be particularly attractive, given the ease of peptide synthesis and modification. A sulfated tetrapeptide that fits this description and binds to vascular endothelial growth factor (VEGF) was discovered using a rationally-designed combinatorial approach. A approximately 6600 member library of tetrapeptides, designed to include heparin functionality, was synthesized by solid-phase Fmoc chemistry. The library was analyzed on-resin for VEGF binding using a fluorescence assay that employed a 7-amino-4-methylcoumarin modified VEGF(165). The beads were ranked according to fluorescent signal and SY(SO(3))DY(SO(3)) was identified as the top binder. The binding affinity of the peptide for VEGF(165) was ascertained by surface plasmon resonance and compared with the heparin mimic suramin; the peptide binds to VEGF(165) 100-fold stronger than the sulfonated compound. These results suggest that the identified peptide may be useful in biomaterial applications where binding of VEGF is desired. PMID- 16701827 TI - Heparin functionalized PEG gels that modulate protein adsorption for hMSC adhesion and differentiation. AB - Heparin was modified with methacrylate groups, copolymerized with dimethacrylated poly(ethylene glycol), and analyzed as a localized delivery vehicle for bFGF and synthetic extracellular matrix for the differentiation of hMSCs. By deriving cues from molecules normally present in the extracellular matrix (ECM), a complex network of collagens, laminin, fibronectin, glycosaminoglycans, and growth factors, synthetic cell scaffolds can be designed that actively sequester important bioactive signals. Among the glycosaminoglycans, heparin binds reversibly with many proteins, therefore, poly(ethylene glycol) based biomaterials, normally resistant to cell adhesion, functionalized with heparin in order to sequester important proteins, can actively and selectively stimulate desired cell functions. Results demonstrate that methacrylate-modified heparin retained its ability to bind heparin-binding proteins both in solution and when copolymerized with dimethacrylated PEG in a hydrogel. In addition, the heparin functionalized gels can deliver biologically active bFGF for up to 5 weeks. Finally, the gels were examined as a potential scaffold for hMSC culture and were found to promote adhesion, proliferation, and osteogenic differentiation. PMID- 16701829 TI - Protrusion force transmission of amoeboid cells crawling on soft biological tissue. AB - We applied a colloidal force microscopy technique to measure the spreading and retraction forces generated by protrusions (pseudopodia) of vegetative amoeboid cells (Dictyostelium discoideum) adhering on soft tissue analogues composed of 2 mm thick hydrogels of hyaluronic acid exhibiting Young's moduli between 10 and 200 Pa. Local shear deformations of the polymer films evoked by magnetic tweezers and by cellular protrusions were determined by analyzing the deflections of colloidal beads randomly deposited on the surface of the polymer cushions, which enabled us to measure forces generated by advancing ("pushing" forces) and retracting ("pulling" forces) protrusions in a direct way. We found that the maximum amplitudes generated by the advancing protrusions (pushes) decrease with increasing stiffness of the HA substrate while the amplitudes of the retractions do not show such a dependence. The maximum forces transmitted by the advancing and retracting protrusions increase with increasing stiffness of the HA films (from 0.02 to 1 nN for the case of pushing). The protrusions spread or retract with constant velocities which are higher for retractions (100 nm s(-1)) than for spreadings (50 nm s(-1)) and are not significantly influenced by the substrate rigidity. We provide evidence that elastic equilibrium during protrusion formation and retraction is maintained by local elastic dipole fields generated at the rim of the protrusions. A model of protrusion force transmission by coupling of growing actin gel in the cytoplasm of the protrusions to cell surface receptors through talin clutches is proposed. PMID- 16701828 TI - Synthesis of biocompatible segmented polyurethanes from aliphatic diisocyanates and diurea diol chain extenders. AB - Many polyurethane elastomers display excellent mechanical properties and adequate biocompatibility. However, many medical-grade polyurethanes are prepared from aromatic diisocyanates and can degrade in vivo to carcinogenic aromatic diamines, although the question of whether the concentrations of these harmful degradation products attain physiologically relevant levels is currently unresolved and strongly debated. It is therefore desirable to synthesize new medical-grade polyurethanes from less toxic aliphatic diisocyanates. In this paper, biocompatible segmented polyurethane elastomers were synthesized from aliphatic diisocyanates (1,4-diisocyanatobutane (BDI) and lysine methyl ester diisocyanate (LDI)), novel diurea diol chain extenders based on tyrosine and tyramine, and a model poly(ethylene glycol) (PEG) diol soft segment. The objectives were to design a hard segment similar in structure to that of MDI-based polyurethanes and also investigate the effects of systematic changes in structure on mechanical and biological properties. The non-branched, symmetric polyurethane prepared from BDI and a tyramine-based chain extender had the highest modulus at 37 degrees C. Introduction of symmetric short-chain branches (SCBs) incorporated in the tyrosine-based chain extender lowered the modulus by an order of magnitude. Polyurethanes prepared from LDI were soft polymers that had a still lower modulus due to the asymmetric SCBs that hindered hard segment packing. Polyurethanes prepared from tyramine and tyrosine chain extenders thermally degraded at temperatures ranging from 110 to 150 degrees C, which are lower than that reported previously for phenyl urethanes. All four polyurethanes supported the attachment, proliferation, and high viability of MG-63 human osteoblast-like cells in vitro. Therefore, the non-cytotoxic chemistry of these polyurethanes make them good candidates for further development as biomedical implants. PMID- 16701830 TI - Bootstrap analysis of the relation between initial adhesive events and long-term cellular functions of human osteoblasts cultured on biocompatible metallic substrates. AB - Classically, the evaluation of cellular adhesion of cells on substrates is limited to the evaluation of cell attachment after some hours. We have claimed for several years that this evaluation is incomplete concerning the evaluation of cell adhesion and more precisely of the quality of the in vitro cell/biomaterial interface. With a view to demonstrating this assertion, we develop in this paper statistical correlations between short-term adhesion (STA) evaluating the attachment after 24 h (IA: initial attachment) and long-term adhesion (LTA) evaluating the strength of the cell/matrix substrate interface over 21 days of culture (AP: adhesion power). Additionally, as the adhesion phase is known to influence further growth of cells we proceed to the correlation of STA with the number of cells after 21 days. We demonstrate statistically the expected positive relation existing between STA and cell growth and we show that this relation is totally independent of the substrate's surface topography or chemistry. More surprisingly, we demonstrate the absence of correlation between IA and AP. This illustrates the fact that different mechanisms underlie STA and LTA. Moreover, this study demonstrates that the evaluation of the number of attached cells after some hours cannot let us presume either that cells will survive or that they will adhere at later times by forming a complex cell/substrate interface by synthesis of extracellular matrix proteins. Finally, the originality of this work lies in the extensive statistical correlation analysis performed between biological parameters describing the cell behaviour on a substrate. PMID- 16701831 TI - Substrate-mediated delivery from self-assembled monolayers: effect of surface ionization, hydrophilicity, and patterning. AB - Gene transfer has many potential applications in basic and applied sciences. In vitro, DNA delivery can be enhanced by increasing the concentration of DNA in the cellular microenvironment through immobilization of DNA to a substrate that supports cell adhesion. Substrate-mediated delivery describes the immobilization of DNA, complexed with cationic lipids or polymers, to a biomaterial or substrate. As surface properties are critical to the efficiency of the surface delivery approach, self-assembled monolayers (SAMs) of alkanethiols on gold were used to correlate surface chemistry of the substrate to binding, release, and transfection of non-specifically immobilized complexes. Surface hydrophobicity and ionization were found to mediate both DNA complex immobilization and transfection, but had no effect on complex release. Additionally, SAMs were used in conjunction with soft lithographic techniques to imprint substrates with specific patterns, resulting in patterned DNA complex deposition and transfection, with transfection efficiencies in the patterns nearing 40%. Controlling the interactions between complexes and substrates, with the potential for patterned delivery, can be used to locally enhance or regulate gene transfer, with applications to tissue engineering scaffolds and transfected cell arrays. PMID- 16701832 TI - A bioactive titanium foam scaffold for bone repair. AB - While titanium has been clinically successful as an orthopedic or dental implant material, performance problems still persist related to implant-bone interfacial strength and mechanical modulus mismatch between titanium and tissue. We describe here the preparation of a titanium foam as a better mechanical match to tissue with surfaces attractive to bone cells through deposition of an organically modified apatite layer (organoapatite). In a rotating bioreactor, these organoapatite-coated foams are successfully colonized by preosteoblastic cells. Finite element analyses suggest that ingrown tissue in these systems may improve both implant performance and tissue formation through load-sharing and stress distribution. The novel metal-ceramic-polymer hybrid materials described here hold great promise for bone tissue engineering. PMID- 16701833 TI - Hydroxyapatite coating of cellulose sponge does not improve its osteogenic potency in rat bone. AB - Regenerated cellulose sponges were coated biomimetically with hydroxyapatite to increase their osteogenic properties. Induction of apatite precipitation was carried out with bioactive glass in simulated body fluid (SBF) for 24 h and the final coating was carried out in 1.5 x concentrated SBF for 14 days. Biomimetically mineralized and non-mineralized sponges were then implanted into standard size femoral cortical defects of rats, and the invasion of bone into the implant was followed up to one year. The apatite coating did not improve the osteoconductive property of cellulose in this rat cortical defect model. In fact, it generated a strong and highly cellular inflammatory reaction and less osteoid tissue. The biomimetic implants contained more immunodetectable TGFbeta1 (a strong stimulator of fibroblast activity) than untreated implants, and also bound more TGFbeta1 in vitro, which could, at least in part, explain the fibrotic invasion of biomimetically mineralized sponges. PMID- 16701834 TI - Rapid and complete cellularization of hydroxyapatite for bone tissue engineering. AB - Using a tissue construct generated by cells in a scaffold in reconstructive surgery, as a substitute for autografts, is still challenging. Routine methods of incorporating cells into scaffolds are either passive, i.e. by gravity, or forced, as in a bioreactor. Extensive use of these methods is obstructed by tissue formation around the scaffold, hindrance in cell penetration and time required for cell coverage within the scaffold. In this study, human osteoblast cells as cell sheet structures were seeded to porous and dense hydroxyapatite with the hypothesis that preservation of native extracellular structures and cell cell contacts would facilitate the cellularization process. Cellularization was assessed by fluorescence, confocal and scanning electron microscopy at intervals of 1 h, 2 days and 7 days. Cell patches with intact cell-cell and cell-extra cellular matrix contact attached and adhered on a scaffold within 1 h. The patches formed a monolayer within 2 days and complete cellularization of the scaffold was attained in 7 days. Cell viability, proliferation and function were assessed to understand the application of cell patch transfer to bone substitute. This novel approach for application in bone tissue engineering was successful in uniform distribution of intact osteoblast cell sheet structures on to bone substitute materials for rapid and complete cellularization without altering material characteristics. PMID- 16701835 TI - Effect of iron on the surface, degradation and ion release properties of phosphate-based glass fibres. AB - Phosphate-based glass fibres (PGF) have the unique characteristic of being completely soluble in an aqueous environment, releasing bioactive and biocompatible ions. They have been proposed as tissue engineering scaffolds for craniofacial skeletal muscle regeneration, where myoblasts are seeded directly onto the fibres. Studies have shown that these cells have a preference in their initial attachment to fibres of certain composition and size, which in turn control the rate of degradation. This study investigated the relationship between the surface properties, degradation properties and ion release (cationic and anionic species) by altering the chemical composition of the PGF. Iron oxide (Fe2O3) was incorporated into glasses containing P2O5 (50 mol%), CaO (30 mol%) and Na2O (20 mol%). Six glass compositions with Fe2O3 ranging from 0 to 5 mol% by replacing the equivalent Na2O mol% were investigated. Contact angle measurements showed that polar interactions occurring on the glass surfaces diminished with increasing Fe2O3 content. This behaviour was reflected in the estimated surface energies of the glasses, where the overall surface energy decreased with increasing Fe2O3 content due to the decrease in polar or acid/base component. The incorporation of up to 5 mol% Fe2O3 into PGF resulted in a significant reduction in the degradation rate (by two orders of magnitude), which can be related to the formation of more hydration resistant P-O-Fe bonds. However, the degradation rate increased with decreasing fibre diameter (comparing average diameters of 31.6 +/- 6.5 microm versus 13.1 +/- 1.3 microm) for a given mass of fibre, and this is related to the surface area to volume ratio. Taken together the results suggest that fibres with the larger diameters and containing 3-5 mol% Fe2O3 could initially be a more durable scaffold than ones with 1 or 2 mol% Fe2O3 for initial cell attachment. PMID- 16701836 TI - Study of the connectivity properties of Bioglass -filled polylactide foam scaffolds by image analysis and impedance spectroscopy. AB - The porous structure of two series of poly(D,L-lactide)/Bioglass composite foams prepared by thermal-induced phase separation was investigated by image analysis and impedance spectroscopy. Polymer solutions of either low or high molecular weight containing different concentrations (up to 50 wt.%) of Bioglass particles of mean particle size d < 5 microm were studied. The morphology of both macro- and micropores was studied by scanning electron microscopy and image analysis of both neat and composite foams (containing 10-50 wt.% Bioglass). The pore connectivity of both neat polymer and composite foams was characterized by impedance spectroscopy in relation with their transport properties. The influence of the foam composition (i.e., polymer molecular weight and concentration of Bioglass on pore microstructure was studied using these non-destructive methods. It was found that addition of Bioglass particles has a pronounced effect on pore orientation, leading to increasing loss of order of pore structure, especially for low-molecular weight PDLLA foams. PMID- 16701837 TI - Design of scaffolds for blood vessel tissue engineering using a multi-layering electrospinning technique. AB - Aiming to develop a scaffold architecture mimicking morphological and mechanically that of a blood vessel, a sequential multi-layering electrospinning (ME) was performed on a rotating mandrel-type collector. A bi-layered tubular scaffold composed of a stiff and oriented PLA outside fibrous layer and a pliable and randomly oriented PCL fibrous inner layer (PLA/PCL) was fabricated. Control over the level of fibre orientation of the different layers was achieved through the rotation speed of the collector. The structural and mechanical properties of the scaffolds were examined using scanning electron microscopy (SEM) and tensile testing. To assess their capability to support cell attachment, proliferation and migration, 3T3 mouse fibroblasts and later human venous myofibroblasts (HVS) were cultured, expanded and seeded on the scaffolds. In both cases, the cell-polymer constructs were cultured under static conditions for up to 4 weeks. Environmental scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), histological examination and biochemical assays for cell proliferation (DNA) and extracellular matrix production (collagen and glycosaminoglycans) were performed. The findings suggest the feasibility of ME to design scaffolds with a hierarchical organization through a layer-by-layer process and control over fibre orientation. The resulting scaffolds achieved the desirable levels of pliability (elastic up to 10% strain) and proved to be capable to promote cell growth and proliferation. The electrospun PLA/PCL bi-layered tube presents appropriate characteristics to be considered a candidate scaffold for blood vessel tissue engineering. PMID- 16701838 TI - Silicon oxycarbide glasses for blood-contact applications. AB - Silicon oxycarbide (SiOxCy) glass compositions are shown to exhibit a variable propensity to contact activate coagulation of whole human blood plasma that depends on X:Y surface stoichiometry. SiOxCy exhibit activation properties similar to pyrolytic carbon (PC) over a broad range of X:Y ratios. Surface composition of SiOxCy glass powders prepared by pyrolysis of thermosetting polysilsequioxanes roughly correlated with total carbon concentration of precursor resins and could be significantly modified by etching in alkaline solutions. Results suggest that SiOxCy may offer unique properties as a substitute for PC in medical-device applications demanding excellent tribological properties, such as artificial heart valves. PMID- 16701840 TI - Localized delivery of nitric oxide from hydrogels inhibits neointima formation in a rat carotid balloon injury model. AB - Using novel nitric oxide (NO)-generating polymeric hydrogels that can be rapidly photopolymerized in situ, we can deliver NO locally at the site of vascular injury. Depending on material design, these poly(ethylene glycol) (PEG)-based hydrogels can generate NO for up to 50 d. This study demonstrates the ability of nitric oxide-generating hydrogels (PEG-Cys-NO) to influence key components of the restenosis cascade both in vitro and in vivo. PEG-Cys-NO hydrogels inhibited smooth muscle cell proliferation, increased endothelial cell proliferation, and inhibited platelet adhesion in vitro. Moreover, in vivo, PEG-Cys-NO hydrogels inhibited intimal thickening in a rat carotid balloon injury model. The perivascular application of NO-generating polymers post-injury reduced neointima formation at 14 d by approximately 80% compared to controls (intimal area/medial area (I/M): PEG-Cys-NO=0.20+/-0.17, control=0.84+/-0.19, p<0.00002; intimal thickness: PEG-Cys-NO=12+/-10 microm, control=60+/-18 microm, p<0.00002). Treatment with the PEG-Cys-NO hydrogels caused a significant decrease in the per cent of proliferating cell nuclear antigen positive medial cells (29+/-5%) at 4 d as compared to treatment with the control hydrogels (51+/-1%, p<0.02). Additionally, vessel re-endothelialization at 14 d was slightly enhanced in the presence of the NO-generating hydrogels. These data indicate that localized delivery of NO from these hydrogels can significantly inhibit neointima formation in a rat carotid balloon injury model and suggest that these materials may be useful in preventing restenosis. PMID- 16701839 TI - Modification of glass fibers to improve reinforcement: a plasma polymerization technique. AB - OBJECTIVES: This study evaluates the effect of plasma treated E-glass fiber to improve the mechanical properties of acrylic resin denture base material, polymethylmethacrlyate (PMMA). Plasma surface treatment of fibers is used as reinforcement in composite materials to modify the chemical and physical properties of their surfaces with tailored fiber-matrix bonding strength. METHODS: Three different types of monomer 2-hydroxyethyl methacrylate (HEMA), triethyleneglycoldimethylether (TEGDME) and ethylenediamine (EDA) were used in the plasma polymerization modification of glass fibers. A radiofrequency generator was used to sustain plasma in a glass vacuum chamber. Glass fibers were modified at the same glow-discharge power of 25 W and exposure time of 30 min for each monomer. Fibers were incorporated into the acrylic with 1% (w/w) loading except control group. Specimens were prepared using a standard mold of 3 cmx0.5 cmx0.8 cm in dimension with eight specimens in each group. Samples were subjected to a flexural strength test set up at a crosshead speed of 5mm/min. Scanning electron microscopy (SEM) was used to examine the microstructure and X-ray photoelectron spectroscopy (XPS) was used for chemical analysis of the surface. RESULTS: Data were analyzed by means of ANOVA and Duncan's tests. Test results revealed that fiber reinforcement had a significant effect on the flexural strength of the specimens (p<0.05). Among the fiber reinforced groups, plasma treatment with EDA monomer resulted in the most significant increase in flexural strength values (p<0.05). XPS results have shown an increasing number of nitrogenous compounds in EDA treated fibers. The chemical structure of the surface, especially with the increase in nitrogenous compounds could give an idea for the amine film deposition and SEM figures showed an increase in surface roughness. SIGNIFICANCE: The results showed that plasma treatment with EDA monomer was an effective alternative method of increasing the flexural strength of PMMA based denture base polymers through fiber reinforcement. PMID- 16701841 TI - Novel capillary channel fiber scaffolds for guided tissue engineering. AB - A novel type of capillary channel fibers (CCFs) containing eight open grooves with depth of 5-15 microm and width of 10 microm were tested for their use in tissue engineering as matrices that provide topographical guidance to neo-tissue development. The matrices fabricated from fibers of poly(l-lactic acid) (PLA) and polyethylene terephthalate (PET) were seeded with rat skin fibroblasts (RSFs) and rat aortic smooth muscle cells (RASMCs) for up to 4 weeks. Cells attached and extended their cytoplasmic lamellapodia within the grooves. The cells were proliferating within the grooves and were highly aligned parallel to the direction of the grooves. RASMCs and RSFs showed highly aligned actin and vimentin cytoskeleton, respectively. The cells also deposited extracellular matrix proteins such as laminin and collagen within the grooves parallel to the groove direction. These CCFs also have the unique ability to move fluids instantaneously by capillary action, thus, have the potential to transport oxygen and nutrients deep within the scaffolds. Such CCF matrices would be useful for creating highly organized tissues such as tendon, ligament, nerve, and cardiac muscle. PMID- 16701842 TI - Dexamethasone loaded bioresorbable films used in medical support devices: structure, degradation, crystallinity and drug release. AB - Bioresorbable polymer films containing dexamethasone (DM) were prepared using a solution processing technique. Investigation of the films focused on cumulative DM release as affected by film morphology (drug location/dispersion in the film) and degradation processes. Two film structures were studied: A-type, a polymer film with large drug crystals located on the film's surface, and B-type, a polymer film with small drug particles and crystals distributed within the bulk. The effect of the polymer's degree of crystallinity on the drug release profile was also studied. Prototypical applications of these films are biodegradable medical support devices which combine mechanical support with drug release. In most of our studied systems the drug release profile from the film is determined mainly by both drug location/dispersion in the film and the polymer's weight loss rate. All release profiles from A-type films exhibited a burst effect of approximately 30%, accompanied by a second release phase at a constant rate, whereas the release profiles from B-type films were determined mainly by the degradation profile of the host polymer, and did not exhibit any burst effect. A high degree of crystallinity is important for the current application, since good mechanical properties are required. This contributes to slower drug release rates, mainly at relatively low weight losses, whereas at high weight losses, where a porous structure is created, the crystallinity almost does not affect the rate of drug release. The shape of the porous structure that develops with degradation also affects the drug release profile from the B-type films. PMID- 16701843 TI - Cell density alters matrix accumulation in two distinct fractions and the mechanical integrity of alginate-chondrocyte constructs. AB - Chondrocyte density in articular cartilage is known to change with the development and growth of the tissue and may play an important role in the formation of a functional extracellular matrix (ECM). The objective of this study was to determine how initial chondrocyte density in an alginate hydrogel affects the matrix composition, its distribution between the cell-associated (CM) and further removed matrix (FRM) fractions, and the tensile mechanical properties of the developing engineered cartilage. Alginate constructs containing primary bovine chondrocytes at densities of 0, 4, 16, and 64 million cells/ml were fabricated and cultured for 1 or 2 weeks, at which time structural, biochemical, and mechanical properties were analyzed. Both matrix content and distribution varied with the initial cell density. Increasing cell density resulted in an increasing content of collagen and sulfated-glycosaminoglycan (GAG) and an increasing proportion of these molecules localized in the CM. While the equilibrium tensile modulus of cell-free alginate did not change with time in culture, the constructs with highest cell density were 116% stiffer than cell free controls after 2 weeks of culture. The equilibrium tensile modulus was positively correlated with total collagen (r2=0.47, p<0.001) and GAG content (r2=0.68, p<0.001), and these relationships were enhanced when analyzing only those matrix molecules in the CM fraction (r2=0.60 and 0.72 for collagen and GAG, respectively, each p<0.001). Overall, the results of this study indicate that initial cell density has a considerable effect on the developing composition, structure, and function of alginate-chondrocyte constructs. PMID- 16701844 TI - Synthesis and characterization of novel biotinylated biodegradable poly(ethylene glycol)-b-poly(carbonate-lactic acid) copolymers. AB - Poly(ethylene glycol)-b-poly(5-benzyloxy-trimethylene carbonate-lactic acid) copolymers (PEG-b-P(BTMC-LA)) were synthesized by ring-opening polymerization of lactide and 5-benzyloxy trimethylene carbonate in the presence of mono-hydroxyl poly(ethylene glycol) with diethyl zinc as catalyst. They were further converted into deprotected copolymers with the pendant hydroxyl groups by hydrogenolysis in the presence of Pd(OH)2/C, and finally conjugated with biotin through the free hydroxyl groups. Gel permeation chromatography, Fourier transform infrared, differential scanning calorimetry and 1H nuclear magnetic resonance studies confirmed the copolymer structures and successful attachment of biotin to the copolymer. PMID- 16701845 TI - Mechanical properties of highly porous PDLLA/Bioglass composite foams as scaffolds for bone tissue engineering. AB - This study developed highly porous degradable composites as potential scaffolds for bone tissue engineering. These scaffolds consisted of poly-D,L-lactic acid filled with 2 and 15 vol.% of 45S5 Bioglass particles and were produced via thermally induced solid-liquid phase separation and subsequent solvent sublimation. The scaffolds had a bimodal and anisotropic pore structure, with tubular macro-pores of approximately 100 microm in diameter, and with interconnected micro-pores of approximately 10-50 microm in diameter. Quasi static and thermal dynamic mechanical analysis carried out in compression along with thermogravimetric analysis was used to investigate the effect of Bioglass on the properties of the foams. Quasi-static compression testing demonstrated mechanical anisotropy concomitant with the direction of the macro-pores. An analytical modelling approach was applied, which demonstrated that the presence of Bioglass did not significantly alter the porous architecture of these foams and reflected the mechanical anisotropy which was congruent with the scanning electron microscopy investigation. This study found that the Ishai-Cohen and Gibson-Ashby models can be combined to predict the compressive modulus of the composite foams. The modulus and density of these complex foams are related by a power-law function with an exponent between 2 and 3. PMID- 16701846 TI - Biodegradable HA-PLA 3-D porous scaffolds: effect of nano-sized filler content on scaffold properties. AB - Scaffolds comprising poly(lactic acid) and nano-hydroxyapatite (HA) were prepared using the solvent-casting/salt-leaching technique. NaCl was used as the leaching agent. Nano-sized HA was synthesized by a hydrothermal method at 170 degrees C and autogenous pressure. High-resolution TEM imaging revealed that the HA particles were ellipsoidal-shaped with needle-like morphologies. The particles had an average size of approximately 25 nm in width and 150 nm in length with aspect ratios ranging from 6 to 8. As the HA content increased in the scaffold from 0 to 50 wt%, the compression modulus of the scaffolds increased from 4.72+/ 1.2 to 9.87+/-1.8 MPa, while the yield strength from 0.29+/-0.03 to 0.44+/-0.01 MPa. Such polymeric scaffolds should be suitable materials for non-load sharing tissue-engineering applications. PMID- 16701847 TI - Photoluminescence of annealed biomimetic apatites. AB - Biomimetic apatite coatings are widely used in orthopaedic applications to provide bioinert material surfaces with bioactive behaviour by means of initiating bone growth at the implant surface. In this study we manufactured biomimetic calcium phosphate coatings consisting of a calcium deficient carbonated apatite by immersing activated titanium platelets into simulated body fluid. The development of the crystal phases was monitored by X-ray diffractometry in addition to Fourier-transform infrared spectroscopy. The microstructure of the biomimetic apatites and phase composition was analysed using scanning and transmission electron microscopy as well as attached energy dispersive X-ray spectrometry. The samples were annealed in air yielding in an inherent luminescence of the biomimetic apatite up to temperatures of 600 degrees C. The photo-induced emission spectra were recorded in the range from 400 to 750 nm at excitation wavelengths ranging 310-450 nm. A blue (437 nm) and a green (561 nm) emission were found between 200 and 600 degrees C visually appearing white. Photoluminescence of annealed biomimetic apatites might be of interest for histological probing and monitoring of bone re-modelling. The results are discussed in terms of chemical and crystallographic changes in the calcium phosphate layer during heat treatment. PMID- 16701848 TI - Modified PMMA cements for a hydrolysis resistant metal-polymer interface in orthopaedic applications. AB - Amongst the many factors influencing the long-term stability of cemented hip prostheses, the interface between the implant and bone cement is considered to be one of the most susceptible to failure. Osteolysis and loosening of the implant can occur by the interaction of mechanically and/or hydrolytically induced bond failure of the metal-cement interface. In this work, an improvement of the hydrolysis resistance of the titanium-bone cement interface was obtained by cement modification with a bifunctional coupling agent combined with a tribochemical TiO2-modification of the metal surface. Methacryloxypropyl trimethoxysilane was added as coupling agent to the PMMA monomer in concentrations between 5 and 20 wt.% followed by the testing the shear bond strength of PMMA/titanium joints before and after ageing in physiological saline solution. It was found that the hydrolysis resistance of the metal-PMMA interface could be significantly improved by the modification of the cement. At the same time, the mechanical properties (compressive and bending strength) of the modified cement were not altered by the addition of the coupling agent. The advantage of the modification of the cement matrix is an easy clinical applicability of the procedure maintaining the processing and implantation techniques of the cement material. PMID- 16701849 TI - New modulated metallic macrocycles: Electrochemistry and their interaction with calf thymus DNA. AB - New modulated pentacoordinate complexes [C17H34N7O2Cu]ClO4 (5), [C17H34N7O2Co]ClO4 (6), [C17H34N7O2Ni] ClO4 (7) have been synthesized by the interaction of 1,8-dihydro-1,3,6,8,10,13 Cu(II) (2), Co(II) (3), Ni(II) (4) hexaazacyclotetradecane complexes and Hsalea N-(2-hydroxy benzyl)-2-amino-1 ethanol ligand (1). All the complexes have been characterized by infrared, electron paramagnetic resonance, 1H and 13C nuclear magnetic resonance (NMR), 2D correlation spectroscopy NMR and ultraviolet-visible (UV-vis) spectroscopy. In all the complexes, the metal center is encapsulated by the Hsalea ligand in a pentacoordinate environment-vis absorption titration and viscometric studies have been carried out to ascertain the interaction of complexes 5 and 6 with calf thymus DNA (CT DNA). The experimental results suggest that complex 5 binds to CT DNA through partial intercalation of the aromatic ring into the base pair of DNA while complex 6 binds binds to CT DNA by electrostatic mode. The intrinsic binding constants K(b) of complex 5 and 6 were found to be 6.8 x 10(-5) M(-1) and 1.8 x 10(-4) M(-1), respectively. The binding of complexes 5 and 6 with CT DNA has also been investigated by cyclic voltammetry. PMID- 16701850 TI - Anti-microbial active composite nanoparticles with magnetic core and photocatalytic shell: TiO2-NiFe2O4 biomaterial system. AB - Reverse micelle and chemical hydrolysis techniques have been successfully combined to synthesize composite nanoparticles consisting of a photocatalytic shell of titania and a magnetic core of nickel ferrite. The nature of titania shell, i.e. anatase or brookite, depends on the TiO2 and NiFe2O4 molar ratio. The work presented here describes the photocatalytic and anti-microbial activity of the composite nanoparticles together with the magnetic characteristics of the nickel ferrite core. The TiO2-coated NiFe2O4 nanoparticles retain the magnetic characteristics of uncoated nanocrystalline nickel ferrites (superparamagnetism; absence of hysteresis, remanence and coercivity at 300 K) encouraging their application as removable anti-microbial photocatalyst nanoparticles that can be extracted from the sprayed surface (human body or environment) after exposure. PMID- 16701851 TI - High strength, low stiffness, porous NiTi with superelastic properties. AB - Near-stoichiometric NiTi with up to 18% closed porosity was produced by expansion at 1200 degrees C of argon-filled pores trapped by powder metallurgy within a NiTi billet. When optimally heat-treated, NiTi with 6-16% porosity exhibits superelasticity, with recoverable compressive strains up to 6% at a maximum compressive stress up to 1700 MPa. The apparent Young's modulus of NiTi with 16% porosity, measured during uniaxial compression, is in the range of 15-25 GPa (similar to human bone), but is much lower than measured ultrasonically (approximately 40 GPa), or predicted from continuum elastic mechanics. This effect is attributed to the reversible stress-induced transformation contributing to the linear elastic deformation of porous NiTi. The unique combination of low stiffness, high strength, high recoverable strains and large energy absorption of porous superelastic NiTi, together with the known biocompatibility of NiTi, makes this material attractive for bone-implant applications. PMID- 16701852 TI - Electrochemical release testing of nickel-titanium orthodontic wires in artificial saliva using thin layer activation. AB - Alloys based on Ni-Ti intermetallics generally exhibit special shape memory and pseudoelastic properties, which make them desirable for use in the dental field as orthodontic wires. The possibility of nickel release from these materials is of high concern, because the allergenicity of this element. The aim of this study was to test pseudoelastic Ni-Ti wires in simulated physiological conditions, investigating the combined effect of strain and fluoridated media: the wires were examined both under strained (5% tensile strain) and unstrained conditions, in fluoridated artificial saliva at 37 degrees C. Real time electrochemical nickel release testing was performed using a novel application of a radiotracer based method, thin layer activation (TLA). TLA was validated, in unstrained conditions, against adsorptive stripping voltammetry methodology. Control tests were also performed in non-fluoridated artificial saliva. From our research it transpired that the corrosion behaviour of Ni-Ti alloy is highly affected by the fluoride content, showing a release of 4.79+/-0.10 microg/cm2/day, but, differently from other biomaterials, it does not seem to be affected by elastic tensile strain. The application of the TLA method in the biomedical field appears a suitable technique to monitor in real time the corrosion behaviour of biomedical devices. PMID- 16701854 TI - Crosslinked polysaccharide nanocapsules: preparation and drug release properties. AB - Crosslinked polysaccharide and composite polysaccharide capsules with diameters ranging from 200 nm to several microns and wall thicknesses of several tens of nanometers have been fabricated by interfacial polymerization of methacrylated N,N-diethylaminoethyl dextran (DdexMA) and DdexMA-vinyl terminated polylactide macromonomers (PLAM). In this method, chloroform droplets or PLAM-containing chloroform droplets were dispersed in water, on which water soluble DdexMA was polymerized to form closed shell structure. Their hollow nature was confirmed by confocal laser scanning microscopy and transmission electron microscopy. Dynamic light scattering revealed that these capsules possess good stability against coagulation during storage. Fourier transform infrared and elemental analysis found that the DdexMA capsules were actually composed of crosslinked DdexMA, while the DdexMA-PLAM capsules were composed of the crosslinked DdexMA-PLAM copolymers and PLAM. By dissolution of ibuprofen in the chloroform droplets, drug loaded capsules were also fabricated. It was found that the loaded drug could be released again in a sustained manner for up to 100 h. The capsule walls had a prominent effect in slowing down the drug release rate, particularly for the DdexMA-PLAM capsules. PMID- 16701853 TI - PEG-based hydrogels as an in vitro encapsulation platform for testing controlled beta-cell microenvironments. AB - An in vitro encapsulation platform for systematically testing the effects of microenvironmental parameters on encapsulated islets was developed. The base encapsulation matrix was a biocompatible hydrogel formed via the photoinitiated polymerization of dimethacrylated poly(ethylene glycol) (PEGDM). The resulting inert encapsulation matrix affords control over the biochemical and biophysical cellular microenvironment and the introduction of systematic changes to this environment. The compatibility of the PEG-based encapsulation platform with pancreatic beta-cells was first established using a murine beta-cell line, MIN6. When cell-cell contacts were introduced via aggregation of MIN6 beta-cells prior to encapsulation, MIN6 beta-cells remained viable within the PEG hydrogel platform throughout 3weeks of in vitro culture. Proliferating cells were observed within encapsulated MIN6 aggregates qualitatively with bromodeoxyuridine staining and quantitatively by measuring the DNA content of encapsulation samples with time. MIN6 beta-cells were encapsulated in hydrogels formed from three PEGDM macromers of varying molecular weights (M (n)=4,000, 8,000, 10,000g/mol), and the resulting differences in hydrogel crosslinking density, which influences transport properties, did not affect encapsulated beta-cell survival. Encapsulated MIN6 beta-cells transplanted into diabetic mice returned blood glucose levels to normal levels, indicating in vivo function. Finally, the compatibility of the PEG encapsulation system with freshly isolated islets was confirmed. PMID- 16701855 TI - Mechanical properties of electrospun fibrinogen structures. AB - Fibrin and fibrinogen have a well-established track record in tissue engineering due to their innate ability to induce improved cellular interaction and subsequent scaffold remodeling compared to synthetic scaffolds. Use of fibrinogen as a primary scaffold component, however, has been limited by traditional processing techniques that render scaffolds with insufficient mechanical properties. The goal of this study was to demonstrate, based on mechanical properties, that electrospun fibrinogen overcomes these limitations and can be successful as a tissue engineering scaffold or wound dressing. Electrospun fibrinogen scaffolds were characterized for fiber diameter and pore area and subsequently tested for uniaxial mechanical properties while dry and hydrated. In addition, uniaxial mechanical testing was conducted on scaffolds treated to regulate scaffold degradation in serum-containing media by supplementing the media with aprotinin or cross-linking the scaffolds with glutaraldehyde vapor. A linear relationship between electrospinning solution concentration and measured fiber diameter was seen; fiber diameters ranged from 120 to 610 nm over electrospinning concentrations of 80 to 140 mg/ml fibrinogen, respectively. Pore areas ranged from 1.3 microm(2) to 13 microm(2) over the same fibrinogen concentrations. Aprotinin in the culture media inhibited scaffold degradation in a predictable fashion, but glutaraldehyde vapor fixation produced less reliable results as evidenced by mechanical property testing. In conclusion, the mechanical characteristics of electrospun fibrinogen strongly support its potential use as a tissue engineering scaffold or wound dressing. PMID- 16701856 TI - Residual monomers and degree of conversion of partially bioresorbable fiber reinforced composite. AB - The aim of this study was to evaluate the total quantity of residual monomer (bis phenyl glycidyl dimethacrylate, i.e. Bis-GMA, and triethylene glycol dimethacrylate, i.e. TEGDMA), residual monomer release into water and the degree of monomer conversion (DC%) of glass fiber-reinforced composites (FRC) with a partially bioresorbable polymer matrix. Another aim was to find out whether the curing mode affects the quantity of residual monomer and degree of conversion. Glass fibers were preimpregnated with a bioresorbable poly(hydroxyproline) amide and non-resorbable Bis-GMA-TEGDMA resin system. Specimens were immersed in water for 1, 3 or 7 days (37 degrees C) to determine the quantity of leached residual monomers, or in the solvent tetrahydrofuran for 3 days to determine the total quantity of residual monomers by high performance liquid chromatography. DC% was measured by Fourier transform infrared spectroscopy. The quantity of residual monomer of the specimens decreased when the specimens contained glass fibers, and/or poly(hydroxyproline) amide, and/or when it was post-cured. The majority of the residual monomers were leached out during the first 24 h of immersion in water. The DC% of the specimens increased when post-cured. Also glass fibers in the composite increased the DC% in contrast to Bis-GMA-TEGDMA resin only. In conclusion, use of poly(hydroxyproline) amide as a sizing of the glass fibers in FRC does not increase the quantity of residual monomers. These results suggest that this new kind of partially bioresorbable FRC has potential for biomedical applications. PMID- 16701857 TI - Nanoscale adhesion, friction and wear studies of biomolecules on silicon based surfaces. AB - Protein layers are deployed over the surfaces of microdevices such as biological microelectromechanical systems (bioMEMS) and bioimplants as functional layers that confer specific molecular recognition or binding properties or to facilitate biocompatibility with biological tissue. When a microdevice comes in contact with any exterior environment, like tissues and/or fluids with a variable pH, the biomolecules on its surface may get abraded. Silicon based bioMEMS are an important class of devices. Adhesion, friction and wear properties of biomolecules (e.g., proteins) on silicon based surfaces are therefore important. Adhesion was studied between streptavidin and a thermally grown silica substrate in a phosphate buffered saline (PBS) solution with various pH values as a function of the concentration of biomolecules in the solution. Friction and wear properties of streptavidin (protein) biomolecules coated on silica by direct physical adsorption and a chemical linker method were studied in PBS using the tapping mode atomic force microscopy at a range of free amplitude voltages. Fluorescence microscopy was used to study the detailed wear mechanism of the biomolecules. Based on this study, adhesion, friction and wear mechanisms of biomolecules on silicon based surfaces are discussed. PMID- 16701858 TI - Scaling effects of wet adhesion in biological attachment systems. AB - Insects have evolved fibrillar attachment devices based on wet adhesion to attach themselves to a variety of surfaces. This paper investigates the scaling effects of wet adhesion mediated by a liquid bridge between a fiber and a solid surface. The influences of liquid volume and contact angles are discussed via a scaling law indicating that the adhesive strength can be enhanced by contact size reduction. Due to the maximum negative pressure in the liquid bridge, there exists a critical length scale at which the system achieves the theoretical tensile strength of the liquid. We conclude that size reduction down to a critical scale results in optimization of the adhesive strength. PMID- 16701859 TI - Mechanisms governing the inelastic deformation of cortical bone and application to trabecular bone. AB - To understand the inelastic response of bone, a three-part investigation has been conducted. In the first, unload/reload tests have been used to characterize the hysteresis and provide insight into the mechanisms causing the strain. The second part devises a model for the stress/strain response, based on understanding developed from the measurements. The model rationalizes the inelastic deformation in tension, as well as the permanent strain and hysteresis. In the third part, a constitutive law representative of the deformation is selected and used to illustrate the coupled buckling and bending of ligaments that arise when trabecular bone is loaded in compression. PMID- 16701860 TI - Selective protein adsorption property and characterization of nano-crystalline zinc-containing hydroxyapatite. AB - Nano-crystalline Zn-containing hydroxyapatite (ZnHAp) was prepared by the wet chemical method and the selective adsorption of essential proteins was examined, taking bovine serum albumin (BSA) and pathogenic protein such as beta(2) microglobulin (beta(2)-MG) as model proteins. Transmission electron microscopy observation and X-ray diffraction analysis indicated that the increase of Zn content led to smaller crystallites and their specific surface area of ZnHAps increased with increasing Zn content, accordingly. Furthermore, the amounts of BSA adsorption on ZnHAp particles decreased with increasing Zn content in spite of the increase in the specific surface area. It is thus concluded that nano crystalline ZnHAps had a highly selective adsorption property with regard to beta(2)-MG. PMID- 16701861 TI - Mineralisation of chitosan scaffolds with nano-apatite formation by double diffusion technique. AB - The study of inorganic crystal assembly in organic matrices has given rise to increasing interest in various fields of materials science to the natural process of biomineralisation. To mimic the formation of hydroxyapatite as natural bone, a double diffusion technique is utilised in this study to nucleate the hydroxyapatite crystals onto three-dimensional porous polymeric scaffolds. The porous polymer scaffolds were produced from chitosan by a thermally induced lyophilisation technique, which yields highly porous, well-controlled anisotropic open pore architecture. The nucleation of hydroxyapatite crystals was initiated at ambient conditions on the surface of the polymer scaffold, which was in contact with a calcium solution chamber, due to diffusion of phosphate ions through the scaffold. The morphology of the mineralised scaffold as analysed by scanning electron microscopy shows that apatite crystals were not only formed on the surface of the scaffold, but also in the pore channels and attached to the pore walls. The X-ray diffraction and Fourier transformed infrared analyses confirmed the phase purity of the formed apatite crystals. The transmission electron microscopy analysis reveals the microstructure of the entangled nano apatite in the chitosan polymeric matrix. The in-vitro cytocompatibility tests with osteoblast-like cells (Saos-2) demonstrated that the biomineralised scaffold is a suitable substrate for cell attachment and migration in bone tissue engineering. PMID- 16701862 TI - Ethoxylated bisphenol dimethacrylate-based amorphous calcium phosphate composites. AB - Improving the anti-demineralizing/remineralizing and mechanical properties of amorphous calcium phosphate (ACP) composites has been the focus of our recent research. In this study, an ethoxylated bisphenol A dimethacrylate (EBPADMA) was blended with triethylene glycol dimethacrylate (TEGDMA), 2-hydroxyethyl methacrylate (HEMA) and methacryloxyethyl phthalate (MEP) to form experimental resins with different EBPADMA/TEGDMA molar ratios (0.50, 0.85 and 1.35) and a constant HEMA/MEP molar ratio (8.26). Composites were prepared by admixture of either unmilled or milled zirconia-ACP filler (40% by mass) and photo-activated resin (60% by mass). One aim was to test if improved ion release can be achieved by elevating the EBPADMA/TEGDMA ratio while lowering the level of surface active methacryloxyethyl phthalate in the resin without adversely affecting the biaxial flexure strength (BFS), degree of vinyl conversion (DC) and water sorption (WS) of composites. A second aim was to assess the effect of using milled vs. unmilled ACP on these properties. Ion release of all composites was significantly above the theoretical minimum necessary for remineralization. Calcium ion release was not impeded by binding with the carboxylic acid groups of MEP. Increased supersaturation was attained with increasing EBPADMA/TEGDMA ratio in the resin. Variations in resin composition had no effect on BFS or DC of composites. The BFS of the milled ACP composites was higher than the BFS of unmilled ACP composites (56% and 79%, respectively for dry and wet specimens). DC of composites was only moderately reduced (13.6% and 7.3%, for unmilled and milled ACP, respectively) compared to unfilled resins. WS decreased in the following order: unmilled ACP composites>milled ACP composites>copolymers. Fine-tuning of the resin and utilizing milled ACP filler improved the remineralizing potential of ACP composites without impeding their DC, BFS or WS. PMID- 16701863 TI - Effects of fibre reinforcement on the mechanical properties of brushite cement. AB - In this study the effect of structure and amount of polyglactin fibre incorporation into a brushite forming calcium phosphate cement system and the effect of mechanical compaction on the fibre modified system were investigated. In comparison the effect of resorbable polycaprolactone surface coating of cement specimens was investigated. The results showed that, apart from the mechanical properties of the reinforcing material, the structure of the incorporated fibres, regular or random, is crucial for the resulting flexural strength and modulus of elasticity. Fibre reinforcement could also be combined with mechanical compaction of the cement/fibre composite paste leading to a possible 7-fold increase in flexural strength or an almost 5-fold increase in modulus of elasticity. Reinforcement of the tensile surface of cement grafts may ultimately improve strength where required, especially in conjunction with bone fixation devices. PMID- 16701864 TI - Mechanical properties of bioactive glass 9-93 fibres. AB - Fibres were manufactured from bioactive glass 9-93 by melt spinning. The manufactured fibres were further characterized by measuring their mechanical properties. The tensile strength of 9-93 glass fibres with a diameter between 20 microm and 140 microm and the flexural strength of glass fibres with a diameter of 500-800 microm were measured. The tensile strength of fibres was highly dependent on fibre diameter. Thin fibres possessed the highest strength, 1,625 MPa, compared to the strength of the thickest fibres tested, which was 617 MPa. The flexural strength of glass 9-93 fibres was approximately 1,000 MPa and the flexural modulus 64 GPa. The Weibull modulus for tensile and flexural strength values was rather low, at about 2-4. PMID- 16701865 TI - The effect of oxalic acid incorporation on the setting time and strength of a glass-ionomer cement. AB - Oxalic acid and its metal oxalate salts have been used extensively in dentistry in a range of applications: as desensitisers, in cavity preparation, and as bonding agents. This study investigated the influence of oxalic acid upon the working time, initial setting time, 24-h hardness and compressive strength of a glass-ionomer cement. Conventional glass-ionomer liquids were prepared from polyacrylic acid, tartaric acid, water, and oxalic acid at concentrations of 0-7% w/w. Liquids were dosed into capsules with a commercial glass-ionomer powder, activated and mixed. The resultant pastes were assessed for working time, initial setting time, 24-h hardness and 24-h compressive strength. Liquids containing 0.5 1% oxalic acid lengthened the working time and initial setting time. At concentrations greater than 2%, both working and initial setting times decreased with increasing oxalic acid. Surface hardness values using liquids with 3% and 7% oxalic acid were less hard than the control. Compressive strength was unchanged over the concentrations tested. Oxalic acid may be a useful reaction modifier in glass-ionomer systems. It accelerated the setting reaction without affecting strength, but was limited to low concentrations because of its relatively poor solubility in water. PMID- 16701866 TI - The effect of fluoride contents in fluoridated hydroxyapatite on osteoblast behavior. AB - Fluoridated hydroxyapatite (FHA) discs with various fluoride contents (0-0.567 mol F(-)/mol) [corrected] have been used to investigate the effect of fluoride content on osteoblastic cell behavior. SAOS-3 rat osteosarcoma cells were cultured on FHA discs for different time periods. The cell behavior was examined in terms of cell attachment, proliferation, morphology and differentiation. The fluoride content in FHA discs strongly affected the cell activities. More cell attachment and proliferation were observed on the fluoride-containing FHA discs than on pure hydroxyapatite (HA). The fluoride content also affected the differentiation behavior of osteoblastic cells. Cells on FHA discs demonstrated a higher alkaline phosphatase (ALP) activity than those on pure HA after 2 [corrected] weeks of culturing. These results suggested that fluoride ions have a significant impact on different osteoblastic cell activities. PMID- 16701867 TI - Influence of fluoride, hydrogen peroxide and lactic acid on the corrosion resistance of commercially pure titanium. AB - Titanium is widely used in dental implantology and orthopaedics due to its excellent corrosion resistance and mechanical properties. However, it has been reported that Ti is sensitive to F(-), H(2)O(2) and lactic acid. Atomic force microscopy (AFM) and scanning electron microscopy (SEM) were used to investigate the corrosion resistance of CP-Ti disks after 9 days immersion in different test solutions, based on artificial saliva containing F(-) (0.5% and 2.5%), H(2)O(2) (0.1% and 10%) and/or lactic acid. Because activated macrophages and bacteria can also release locally some of these oxidative compounds, we investigated the role of these cells when plated onto titanium disks. The surface roughness (R(a)) was highly increased when titanium disks were immersed in artificial saliva containing F(-), H(2)O(2) and lactic acid. After 21 days of cell culture, R(a) was significantly increased on disks incubated with activated-J774.2 cells or Streptococcus mitis. AFM appeared to be more sensitive than SEM in evaluating the corrosion of the titanium. Chemical species, either environmental or those released by macrophages and bacteria, can provoke a marked attack of the titanium surface. PMID- 16701868 TI - The effect of hyaluronic acid and phospholipid based lubricants on friction within a human cartilage damage model. AB - The lubricating abilities of different formulations of high molecular weight hyaluronic acid (HA), dipalmitoyl phosphatidylcholine (DPCC) and mixtures of both HA and DPCC were assessed in an in vitro model. Levels of start-up friction were determined using an osteoarthritis (OA) damaged human cartilage model set within a specially designed friction rig. To examine the long term benefits of HA, the extent of penetration of HA into cartilage tissue was investigated using fluorescently labelled HA and confocal microscopy. It was found that in this model, all formulations of HA and the majority of DPCC lubricants reduced friction (HA 5 and 10 mg ml(-1), DPPC 200 mg ml(-1) reductions of 51.9%, 46.7% and 46.5% respectively), compared to a Ringers solution control. Lubrication was found not to be concentration dependent for HA formulations, but concentration was key for DPCC lubrication (100 mg ml(-1) reduced friction by only 15.9%). By combining HA and DPCC (HA/DPPC; 5 mg ml(-1)/100 mg ml(-1) and 10 mg ml(-1)/200 mg ml(-1)), a further improvement was noted (69.5% and 61.9%, respectively) as the mean levels of friction were reduced by up to a further 17% than the most effective individual formulation (HA 5 mg ml(-1)). Penetration of HA into bovine cartilage by up to 300 microm from the surface was observed over a 48 h period. It was observed that HA specifically targeted the chondrocytes as it was primarily found within the lacunae surrounding the cells. PMID- 16701869 TI - In situ observation of biomolecules patterned on a PEG-modified Si surface by scanning probe lithography. AB - A Si(100) wafer was modified with methoxy-poly(ethylene glycol) (M-PEG silane) via a self-assembly technique and nano-/micro-sized patterns were then fabricated by scanning probe lithography. The protrusive silicon dioxide pattern was more reactive compared to the non-patterned area, i.e. the PEG deposited area. To demonstrate the feasibility of the submicron patterning of protein based on the anodic oxidation of the Si surface by atomic force microscopy (AFM), streptavidin labelled with Au-colloidal particle and non-labelled streptavidin were site selectively immobilized on the patterned areas. The streptavidin-patterned surface produced by these procedures can be utilized for the detection of biotinylated materials, such as an antibody and an antigen. A patterned silicon surface is the basis of biosensing devices, in which the patterned areas serve as sensing elements that directly interact with bioanalytes, while the background of the substrate remains passive to the deposition of analytes, thus resulting in a high signal-to-noise ratio. PMID- 16701871 TI - Functionally graded bioactive coatings: reproducibility and stability of the coating under cell culture conditions. AB - This work sought to provide a basic protocol for treatment of functionally graded bioactive glass coatings (FGC) that reliably adhere to titanium alloy (Ti6Al4V) prior to in vivo evaluation. The effect of the fabrication process on glass structure and reproducibility of the coating's properties, and the effect of cell culture conditions on the integrity of the coating were assessed. The structure of FGCs was compared to that of the as cast glass used as a top coating. X-ray diffraction (XRD) showed that the fabrication process resulted in 5.9+/-3.0 vol.% crystallization, while glass as cast was amorphous. Glass as cast and coatings behaved similarly in simulated body fluid (SBF): an amorphous layer rich in phosphate formed, and it crystallized, over 4 weeks, into apatite-like mineral (Fourier transform infrared spectroscopy (FTIR), XRD, scanning electron microscopy (SEM)). Reproducibility of the fabrication process was tested from three batches of coatings by measuring thickness and crystallinity. MC3T3-E1.4 mouse pre-osteoblast cells were cultured and induced to mineralize on FGCs, either as made or pre-conditioned in SBF. The sub-surface glass silica network in FGCs was compromised by cell culture conditions. A crystalline phosphate was formed during pre-conditioning (XRD, FTIR, and SEM). SBF-pre-conditioning stabilized the coatings. Thus incubation in SBF is recommended to produce a stable coating. PMID- 16701870 TI - Pre-B-cell colony-enhancing factor (PBEF/Visfatin) gene expression is modulated by NF-kappaB and AP-1 in human amniotic epithelial cells. AB - A localized intrauterine inflammatory response is often associated with the initiation of normal human parturition, whereas infection causes a similar but more florid response initiating preterm labor. Pre-B-cell colony-enhancing factor (PBEF) is expressed in the human fetal membranes and is up-regulated by labor, severe infection and inflammatory stimuli. The aim of this study was to determine the involvement of the transcription factors NF-kappaB and AP-1 in the response of PBEF to an inflammatory stimulus and compare it with IL-8. The results showed that this treatment of amniotic epithelial-like cells (WISH) and primary amniotic epithelial cells increased expression of PBEF and IL-8, but IL-8 responded 100 fold more than PBEF. IL-1beta treatment together with a panel of NF-kappaB and AP 1 inhibitors demonstrated the involvement of these transcription factors in the up-regulation of PBEF. These data show that an inflammatory stimulus in the fetal membranes inducing NF-kappaB and AP-1 would up-regulate PBEF as well as IL-8. PMID- 16701872 TI - Surface characterisation of various bone cements prepared with functionalised methacrylates/bioactive ceramics in relation to HOB behaviour. AB - This study reports the relationship between the biocompatibility and surface properties of experimental bone cements. The effect of hydroxyapatite (HA) or alpha-tri-calcium phosphate (alpha-TCP) incorporated into bone cements prepared with methyl methacrylate as base monomer and either methacrylic acid or diethyl amino ethyl methacrylate (DEAEMA) as comonomers was investigated. The in vitro biocompatibility of these composite cements was assessed in terms of the interaction of primary human osteoblasts grown on the materials over a period of 5 days and compared with a control surface. These results were related to the surface properties investigated through a number of techniques, namely Fourier transform infrared, contact angle measurements, X-ray photoelectron spectroscopy and energy dispersive analysis of X-rays. Complementary techniques of thermal analysis and ion chromatography were also performed. Biocompatibility results showed that the addition of alpha-TCP improves biocompatibility regardless of comonomer type. This is in contrast to HA-based cements where cell proliferation was significantly lower. Surface characterisations showed that structural integrity of the materials was maintained in the presence of the acid and base comonomers, and water contact angles were reduced particularly in DEAEMA containing materials. Furthermore, ion chromatography confirmed higher Ca2+ and PO4(3-) ion release by both types of ceramics, particularly for those containing DEAEMA. In conclusion, the incorporation of acidic and basic comonomers to either HA or alpha-TCP ceramics containing bone cements can have differential effects upon the attachment and proliferation of bone cells in vitro. Moreover, those cements consisting of alpha-TCP and containing DEAEMA comonomer indicated the most favourable biocompatibility. PMID- 16701874 TI - Fouling and non-fouling surfaces produced by plasma polymerization of ethylene oxide monomer. AB - This paper presents the results of plasma polymerization using diethylene glycol dimethyl ether as a precursor in a capacitively coupled radio frequency system. The chemical structure of the coatings was characterized using several analysis techniques (X-ray photoelectron spectroscopy, Fourier transform-infrared spectroscopy, ellipsometry), while the biological response of these coatings has been tested by protein adsorption and cell culture experiments. The modulation of the input plasma power controls the concentration of polyethylene oxide groups in the coatings and allows the production of films with opposite protein and cell repellent properties. The study of the stability of these coatings in different media (water, acetone, phosphate-buffered saline) reveals that these films could be involved in classical lift-off processes for the production of patterned surfaces. PMID- 16701875 TI - Growth of hydroxyapatite in a biocompatible mesoporous ordered silica. AB - A novel biomaterial (HA-SBA-15) has been developed based on the growth of calcium phosphate hydroxyapatite (HA) nanoparticles within an organized silica structure (SBA-15). Characterization of the material was carried out using a combination of X-ray diffraction, X-ray fluorescence, transmission electron microscopy, N2 adsorption-desorption isotherms and nuclear magnetic resonance. Transmission electron microscopy observations and N2 porosimetry revealed the crystallization of hydroxyapatite nanoparticles inside the mesopore cavities of the silica structure. Specific surface areas of 760 m2 g(-1) and 260 m2 g(-1) were measured for the SBA-15 and the HA-SBA-15 material, respectively. The hydroxyl groups present in the silica nanostructure surface have brought about cationic defects in the silicium sites, mainly with those of tetrahedral symmetry, and promoted the formation of siloxanes. 29Si MAS-NMR analysis shows a significant reduction of the silanol groups concentration with HA growing within the base (SBA-15) material. Studies and brain tissue biocompatibility tests were carried out. Histopathological studies on the SBA-15 implant material showed no changes to the tissue nearby. The results confirmed the synthesis of a silica-based composite containing HA nanoparticles with the potential for biomedical applications. PMID- 16701873 TI - Surface modification of poly(L-lactic acid) to improve its cytocompatibility via assembly of polyelectrolytes and gelatin. AB - Poly(L-lactide) (PLLA) surface was modified via aminolysis by poly(allylamine hydrochloride) (PAH) at high pH and subsequent electrostatic self-assembly of poly(sodium styrenesulfonate) (PSS) and PAH, and the process was monitored by X ray photoelectron spectroscopy (XPS) and contact angle measurement. These modified PLLAs were then used as charged substrates for further incorporation of gelatin to improve their cytocompatibility. The amphoteric nature of the gelatin was exploited and the gelatin was adsorbed to the negatively charged PLLA/PSS and positively charged PLLA/PAH at pH=3.4 and 7.4, respectively. XPS and water contact angle data indicated that the gelatin adsorption at pH=3.4 resulted in much higher surface coverage by gelatin than at pH=7.4. All the modified PLLA surfaces became more hydrophilic than the virgin PLLA. Chondrocyte culture was used to test the cell attachment, cell morphology and cell viability on the modified PLLA substrates. The results showed that the PAH and PSS modified PLLA exhibited better cytocompatibility than virgin PLLA, and the incorporation of the gelatin on these modified PLLA substrates further improved their cytocompatibility, with the PLLA/PSS substrate treated with the gelatin at pH=3.4 being the best, exceeding the chondrocyte compatibility of the tissue culture polystyrene. PMID- 16701876 TI - Preparation of SBF with different HCO3- content and its influence on the composition of biomimetic apatites. AB - The bioactivity of bone and dental implant materials is usually tested in vitro using simulated body fluid (SBF). The composition of common SBF differs from that of blood plasma in that it has a higher Cl- and a lower HCO3- concentration, which affects the composition of in vitro formed bone-like apatite. Five different SBFs with a composition of 142 Na+, 5 K+, 2.5 Ca2+, 1 Mg2+, 1SO4(2-), 1HPO4(2-), and 136 (Cl-+HCO3-) mmol/l were prepared with HCO3- concentrations ranging from 5 to 27 mmol/l. The SBF solutions were prepared by mixing stable concentrated solutions, which increase the reproducibility of in vitro tests due to negligible changes of pH during preparation. The high stability of thus prepared SBF enables the evaluation of hydroxyapatite formation on the surface of bioactive materials without the negative effect of spontaneous precipitation. Furthermore, the use of concentrated solutions offers a facile way to prepare SBF with different ionic contents and thus modify the composition of Ca-P layers precipitated on the surface of the bioactive materials exposed to the SBF solutions. The SBF solutions were shown to be supersaturated with respect to slightly carbonated apatite. The Fourier transform infrared (FT-IR), Raman and X ray analyses of the precipitated layers indicate that the HCO3- content in SBF influences the composition and structure of the calcium phosphates obtained. It can be supposed that as long as the HCO3- concentration in the testing solutions is lower than 20 mmol/l, only B-type HCA precipitates. At higher HCO3- concentrations, it can be assumed that A-type HCA forms as well considering FT IR, Raman and X-ray measurements. PMID- 16701877 TI - Rabbit pilot study on the resorbability of three-dimensional bioactive glass fibre scaffolds. AB - Bioactive glass composed of Na2O-K2O-MgO-CaO-B2O3-P2O5-SiO2 is used in this study to manufacture three-dimensional glass fibre scaffolds for a synthetic bone filler material for the treatment of bone defects. The glass is characterized by a large working range, which is the temperature interval at which forming of glass can take place. A preliminary in vivo study on New Zealand skeletally mature rabbit's tibia is reported here. Bone defects were prepared in the medial surfaces of the diaphyses of the tibia. For the first time melt derived three dimensional bioactive glass fibre constructs were used to fill the cavities. The different implants investigated here were a scaffold with a porosity of 45-50%, scaffold with a porosity of 55-60% and morsels with a porosity of 55-60%. The implanted bone substitutes were dissected after 6 months and evaluated by histological and synchrotron radiation micro tomography analysis. PerioGlas and empty defects were used as positive and negative controls, respectively. The result was that the surgically created tibial defects were healed and new bone formation was found in the medullary cavities. Despite the intrinsic limitations of a pilot study, the preliminary results indicate that in 6 months the glass fibre scaffolds are completely resorbed and that the osteoconductive properties of the filling material are strictly correlated with the structural and morphological characteristics of the bone substitute. PMID- 16701878 TI - Production of ultra-fine bioresorbable carbonated hydroxyapatite. AB - Ionic-substituted hydroxyapatite (HAp) based materials may be a better choice than pure HAp owing to their similarity in chemical composition with biological apatite. The present study reports a process for the production of carbonated hydroxyapatite (CHAp) using microwaves. The CHAp was evaluated for its phase purity, chemical homogeneity, functionality, morphology, and solubility. The CHAp thus obtained was compared with a pure HAp and a biological apatite, which provides quite an interesting insight into the carbonate substitution. The in vitro ionic dissolution rates determined under physiological conditions clearly demonstrate the soluble nature of CHAp compared to HAp. The overall results indicate that the processed CHAp has increased resorption relative to pure HAp and has a chemical composition corresponding to some extent with that of biological apatite. PMID- 16701879 TI - Novel block copolymer (PPDO/PLLA-b-PEG): enhancement of DNA uptake and cell transfection. AB - The cationic lipid mediated uptake of plasmid DNA by cells in monolayer culture was significantly enhanced with an aqueous solution of the block copolymer poly(p dioxanone-co-L-lactide)-b-poly(ethylene glycol) (PPDO/PLLA-b-PEG). Plasmid uptake studies with DNA encoding the beta-galactosidase gene and cytotoxicity evaluations were performed on MCF-7, NIH 3T3 and CT-26 cell lines. Transfection yields and time courses for maximum release of FITC labeled DNA in MCF-7 cells were observed and quantified by beta-galactosidase assay and spectrofluorometry, respectively. The reported results suggest that the studied block copolymer might be useful for the enhancement of polycation mediated transfection and could find application in gene therapy. PMID- 16701880 TI - Biochemical and mechanical behavior of ostrich pericardium as a new biomaterial. AB - We have performed a comparative analysis of glutaraldehyde-preserved ostrich pericardium, as a novel biomaterial, with bovine pericardium. The biochemical characteristics (histology, water content, amino acid composition, and collagen and elastin contents), mechanical properties, and in vivo calcification in a subcutaneous rat model were examined. Ostrich pericardium is slightly thinner and shows a higher water content (70+/-2% vs. 62+/-2%) than bovine pericardium. Additionally, ostrich pericardium presents 1.6-fold lower elastin content and a lower percentage of collagen in reference to the total protein content (68+/-2% vs. 76+/-2%). However, ostrich pericardium shows better mechanical properties, with higher tensile stress at rupture (32.4+/-7.5 vs. 11.5+/-4.6) than calf pericardium. In vivo calcification studies in a rat subcutaneous model show that ostrich pericardium is significantly less calcified than bovine pericardium (23.95+/-13.30 vs. 100.10+/-37.36 mg/g tissue) after 60 days of implantation. In conclusion, glutaraldehyde-stabilized ostrich pericardium tissue shows better mechanical properties than calf tissue. However, calcium accumulation in implanted ostrich tissue is still too high to consider it a much better alternative to bovine pericardium, and anticalcification treatments should be considered. PMID- 16701881 TI - Biocompatibility and function of microencapsulated pancreatic islets. AB - Encapsulation of pancreatic islets in alginate is used to protect against xenogenic rejection in different animal models. In this study, several factors, including differences in alginate composition, the presence or absence of xenogenic islet tissue and a transient immunosuppression, were investigated in a model of bovine islet transplantation in rats. A pure alginate with predominantly guluronic acid (Manugel) and an ultrapure low viscosity guluronic acid alginate (UP-LVG) were used. When microcapsules of Manugel or UP-LVG containing 16,000 bovine islet equivalents were transplanted in diabetic rats, we observed normoglycemia for 8.3+/-0.7 (range 6-12 days) and 7.5+/-0.2 days (range 7-8 days) on average, respectively. To ameliorate immunoprotection of alginate microcapsules we repeated the same experiments using transient immunosuppressive therapy. Low doses of cyclosporin A (CyA) administered for 18 days after implantation increased the time in normoglycemia, which averaged 27+/-3 days (range 8-55 days) in Manugel capsules while in UP-LVG capsules it averaged 18+/-8 days (range 3-39 days). The surface of recovered capsules showed less capsules free of overgrowth in Manugel with respect to UP-LVG alginate. These data were comparable with those observed in empty microcapsules similarly implanted, indicating that the capsular overgrowth was not promoted by the presence of xenogenic islet tissue. In recovered Manugel capsules the percentage of capsules without fibrotic overgrowth was higher than that observed without CyA. The same observation was made in empty capsules. These observations indicate that a combination of a highly purified alginate and short-term immunosuppression prolong islet function in a model of xenotransplantation. PMID- 16701882 TI - Bridging the gap--biocompatibility of microelectronic materials. AB - There is an increasing interest in cell-based microelectronic biosensors for high throughput screening of new products from the biotech pipeline. This requires fundamental knowledge of the biocompatibility of the materials used as the growing support for the cells. Using monolayer-forming Caco-2 cells of human origin, the biocompatibility of silicon wafers coated with various metals, dielectrics and semiconductors was assessed. Besides microscopic inspection, proliferation of cells indicating viability as well as brush border enzyme activity indicating differentiation of adherent growing cells were chosen as parameters to estimate biocompatibility. The type of wafer used for deposition of the coating initially influences the biocompatibility of the final product. Whereas p-doped silicon was fully biocompatible, n-doped silicon reduced the proliferation of cells. Among the different coatings, Al and Ti even increased the cell growth as compared to glass. Culturing the cells for 6 days on coated wafers demonstrated that the differentiation of adhering cells on Ti- and ZrO2 coated wafers was comparable to glass, whereas coatings with Si3N4, Au, Al, and ITO reduced differentiation to 15-35%. In the cases of Au and Si3N4 this effect equilibrated with prolonged culturing. These results demonstrate the importance of a careful selection of the materials used for the production of cell-based biosensors. PMID- 16701883 TI - Aberrant DNA demethylation in promoter region and aberrant expression of mRNA of PAX4 gene in hematologic malignancies. AB - The PAX4 gene, a member of the paired box (PAX) gene family, is thought to be involved in regulating the fate of beta-cells in the mammalian pancreas. We observed the aberrant expression of PAX4 mRNA in 10 of 15 hematologic cell lines analyzed by RT-PCR. The restoration of PAX4 gene expression after treatment with the demethylating agent 5-aza-2'-deoxycitidine, as well as bisulfite sequencing analysis, indicated that gene overexpression was caused by DNA demethylation at the promoter region. Such DNA demethylation also was observed in primary lymphoma (20 out of 45 patients) on combined bisulfite restriction assay (COBRA). Forced expression of the PAX4 gene in the HEK293 and SHSY/610 cell lines conferred positive effects on cell growth. This profile of PAX4 thus corresponds to that of a candidate oncogene in hematologic malignancies. PMID- 16701884 TI - Physical matrices stabilized by enzymatically sensitive covalent crosslinks. AB - This work describes a unique system of gel and gel-like materials formed from physical bonds between heparin and heparin binding peptides (dG-PBD) coupled to multivalent poly(ethylene glycol) vinyl sulfone star polymers (PEGVS) and formed from covalent bonds between an enzymatically sensitive crosslinker and PEGVS. Dynamic mechanical testing revealed that the viscoelastic behavior and gelation kinetics of 10% (w/v) gels formed from 2:1 dG-PBD:PEGVS, heparin, and crosslinker (2:1 gels) and from 3:1 dG-PBD:PEGVS, heparin, and crosslinker (3:1 materials) were significantly influenced by the presence of both physical and covalent bonds. Furthermore, the mechanical properties of both compositions were thermally responsive and reversible. At 4 degrees C, the storage modulus, G', for 2:1 gels (5005.3+/-592.0Pa) and 3:1 materials (5512.0+/-272.7Pa) were statistically similar; however, at 45 degrees C, G' of 2:1 gels decreased to 477.9+/-150.4Pa, and the viscoelastic behavior of 3:1 materials was dominated by viscous behavior. In addition, the mechanical properties of 2:1 gels and 3:1 materials were sensitive to the frequency of the applied stress at 4 degrees C, 20 degrees C, and at 37 degrees C. Although the covalent bonds within the materials provided mechanical stability, the overall viscoelastic response of this system could be dominated by physical crosslinks under certain conditions. Results from degradation studies indicated that the crosslinker was sensitive to collagenase type I but not to thrombin or heparinase I, and a hemolysis assay suggested that the 2:1 gels and 3:1 materials might be biocompatible. These materials could be useful to study the role of physical interactions within networks that mimic the extracellular matrix. PMID- 16701885 TI - Collective effects in cellular structure formation mediated by compliant environments: a Monte Carlo study. AB - Compliant environments can mediate interactions between mechanically active cells like fibroblasts. Starting with a phenomenological model for the behavior of single cells, we use extensive Monte Carlo simulations to predict non-trivial structure formation for cell communities on soft elastic substrates as a function of elastic moduli, cell density, noise and cell position geometry. In general, we find a disordered structure as well as ordered string-like and ring-like structures. The transition between ordered and disordered structures is controlled both by cell density and noise level, while the transition between string- and ring-like ordered structures is controlled by the Poisson ratio. Similar effects are observed in three dimensions. Our results suggest that in regard to elastic effects, healthy connective tissue usually is in a macroscopically disordered state, but can be switched to a macroscopically ordered state by appropriate parameter variations, in a way that is reminiscent of wound contraction or diseased states like contracture. PMID- 16701886 TI - Two photon induced polymerization of organic-inorganic hybrid biomaterials for microstructured medical devices. AB - Three-dimensional microstructured medical devices, including microneedles and tissue engineering scaffolds, were fabricated by two photon induced polymerization of Ormocer organic-inorganic hybrid materials. Femtosecond laser pulses from a titanium:sapphire laser were used to break chemical bonds on Irgacure 369 photoinitiator within a small focal volume. The radicalized starter molecules reacted with Ormocer US-S4 monomers to create radicalized polymolecules. The desired structures are fabricated by moving the laser focus in three dimensions using a galvano-scanner and a micropositioning system. Ormocer surfaces fabricated using two photon induced polymerization demonstrated acceptable cell viability and cell growth profiles against B35 neuroblast-like cells and HT1080 epithelial-like cells. Lego-like interlocking tissue engineering scaffolds and microneedle arrays with unique geometries were created using two photon induced polymerization. These results suggest that two photon induced polymerization is able to create medical microdevices with a larger range of sizes, shapes, and materials than chemical isotropic etching, injection molding, reactive ion etching, surface micromachining, bulk micromachining, polysilicon micromolding, lithography-electroforming-replication, or other conventional microfabrication techniques. PMID- 16701887 TI - Chemical synthesis of poly(lactic-co-glycolic acid)/hydroxyapatite composites for orthopaedic applications. AB - Hydroxyapatite-biodegradable polymer composites were synthesized by a colloidal non-aqueous chemical precipitation technique at room temperature. The starting materials used for synthesizing hydroxyapatite (HA, Ca(10)(PO(4))(6)(OH)(2)) were Ca(NO(3))(2) x 4H(2)O and H(3)PO(4), resulting in single phase HA while poly(d,l lactic-co-glycolic acid) (PLGA) was used as the biodegradable polymer component. The composites were prepared containing 10, 20, and 30 wt.% HA in the presence of the dissolved polymer without evidence of any visible phase separation of the particulates from the PLGA polymer. In addition, the pH changes occurring in the solution during precipitation, the yield of the ceramic due to the chemical reaction, bonding characteristics between the ceramic and the polymer, the microstructure, tensile strength, and thermal stability of the composites have been investigated. Additional in vitro studies include osteoblast-like adhesion assessment on composites utilizing MG63 cells. The results of these studies are described and discussed. PMID- 16701888 TI - Hydrolytic degradation of electron beam irradiated high molecular weight and non irradiated moderate molecular weight PLLA. AB - The purpose of this study is to examine the hydrolytic degradation of electron beam irradiated ring-opening polymerized (ROP) poly(l-lactide) (PLLA-ir) and non irradiated melt polycondensation polymerized poly(l-lactic acid) (PLLA-pc). It was observed that irradiation increases the hydrolytic degradation rate constant for ROP PLLA. This was due to a more hydrophilic PLLA-ir, as a result of irradiation. The degradation rate constants (k) of PLLA-ir samples were also found to be similar, regardless of the radiation dose, and an empirically formulated equation relating hydrolytic degradation time span to radiation dose was derived. The k value for PLLA-pc was observed to be lower than that for PLLA ir, though the latter had a higher molecular weight. This was due to the difference in degradation mechanism, in which PLLA-ir undergoes end group scission, through a back- biting mechanism, during hydrolysis and thus a faster hydrolysis rate. Electron beam irradiation, though accelerates the degradation of PLLA, has been shown to be useful in accurately controlling the hydrolytic time span of PLLA. This method of controlling the hydrolytic degradation time was by far an easier task than through melt polycondensation polymerization. This would allow PLLA to be used for drug delivery purposes or as a temporary implant that requires a moderate time span (3-6 months). PMID- 16701890 TI - Chitosan scaffolds: interconnective pore size and cartilage engineering. AB - This study was designed to determine the effect of interconnective pore size on chondrocyte proliferation and function within chitosan sponges, and compare the potential of chitosan and polyglycolic acid (PGA) matrices for chondrogenesis. Six million porcine chondrocytes were seeded on each of 52 prewetted scaffolds consisting of chitosan sponges with (1) pores 10 microm in diameter (n=10, where n is the number of samples); (2) pores measuring 10-50 microm in diameter (n=10); and (3) pores measuring 70-120 microm in diameter (n=10), versus (4) polyglycolic acid mesh (n=22), as a positive control. Constructs were cultured for 28 days in a rotating bioreactor prior to scanning electron microscopy (SEM), histology, and determination of their water, DNA, glycosaminoglycan (GAG) and collagen II contents. Parametric data was compared (p=0.05) with an ANOVA and Tukey's Studentized range test. PGA constructs consisted essentially of a matrix containing more cells than normal cartilage. Whereas very few remnants of PGA remained, chitosan scaffolds appeared intact. DNA and GAG concentrations were greater in PGA scaffolds than in any of the chitosan groups. However, chitosan sponges with the largest pores contained more chondrocytes, collagen II and GAG than the matrix with the smallest pores. Constructs produced with PGA contained less water and more GAG than all chitosan groups. Chondrocyte proliferation and metabolic activity improved with increasing interconnective pore size of chitosan matrices. In vitro chondrogenesis is possible with chitosan but the composition of constructs produced on PGA more closely approaches that of natural cartilage. PMID- 16701889 TI - Pseudo-dry-spinning of chitosan. AB - A pseudo-dry-spinning process of chitosan without any use of organic solvent or cross-linking agent was studied. A highly deacetylated chitosan (degree of acetylation=2.7%) from squid-pens, with a high weight-average molecular weight (M(W)=540,000 g/mol) was used. The polymer was dissolved in an acetic acid aqueous solution in order to obtain a polymer concentration of 2.4% w/w with a stoichiometric protonation of the -NH(2) sites. The coagulation method consisted of subjecting the extruded monofilament to gaseous ammonia. The alkaline coagulation bath classically used in a wet-spinning process was therefore not useful. A second innovation dealt with the absence of any aqueous washing bath after coagulation. The gaseous coagulation was then directly followed by a drying step under hot air. When the chitosan monofilament coagulated in the presence of ammonia gas, ammonium acetate produced with the fiber could be hydrolyzed into acetic acid and ammonia, easily eliminated in their gaseous form during drying. The pseudo-dry-spinning process did not give rise to any strong degradation of polymer chains. After 2 months at ambient atmosphere, chitosan fibers could then be stored without any significant decrease in the M(W), which remained at a rather high value of 350,000 g/mol. The obtained chitosan fibers showed a smooth, regular and uniformly striated surface. PMID- 16701891 TI - Influence of casting procedures on the corrosion resistance of clinical dental alloys containing palladium. AB - The aim of this study was to compare the in vitro corrosion resistance in artificial saliva of two palladium-silver alloys (a Pd-Ag (Pors on 4) and an Ag Pd (Palliag LTG)), with and without casting defects; 1 nickel-chrome alloy and 1 high-gold alloy, cast under recommended conditions, served as controls. For each of the palladium-based alloys, three specimens corresponding to three different casting conditions were used: under recommended conditions, with the use of a graphite-containing investment and crucible, and by reusing the sprues and sprue button. The electrochemical tests were run in Fusayama-Meyer artificial saliva. The open-circuit potential was recorded in mV/SCE at t=24h. Then, potentiodynamic polarization was performed to measure the polarization resistance (R(p)) in kOmega cm(2) and the corrosion current (i(corr)) in microA cm(-2). Data were evaluated with one-way analysis of variance and multiple comparisons test (alpha=0.05). In addition, each specimen was examined by scanning electron microscopy. Compared to the control alloys, the electrochemical experiments in artificial saliva indicated satisfactory corrosion resistance for the Pd-Ag and Ag-Pd alloys; these results are related to their high noble metal content and stable substructure. The Pd-Ag alloy displayed superior electrochemical properties to those of the Ag-Pd alloy regardless of the casting condition. The use of the graphite-containing crucible and investment during the cast process did not dramatically reduce the corrosion resistance values, but the reuse of sprues and the sprue button did. The optimal corrosion resistance values were obtained for the alloys cast according to the recommended conditions. PMID- 16701892 TI - Hydroxyapatite/SiO(2)-CaO-P(2)O(5) glass materials: in vitro bioactivity and biocompatibility. AB - Materials obtained by the heat treatment of mixtures of hydroxyapatite (HA) and a silicate-based glass of the system SiO(2)-CaO-P(2)O(5) have been investigated. The influence of the glass content on the porosity, microstructure and on the constituent phases of the final materials was studied. The influence of these factors on the in vitro bioactive behaviour of the obtained materials was also investigated. In addition, an in vitro biocompatibility assay with osteoblastic like cells was carried out. The addition of the glass to HA induced different solid-state reactions that yield the transformation of HA into alpha- and beta tricalcium phosphate as well as the formation of silicon-containing phases (silicocarnotite or pseudowollastonite). In these mixtures an enhancement in the porosity, pore size and a heterogeneous microstructure was observed, compared with the precursors. As the sol gel glass content increased, the previous effects were higher. The materials showed the formation of an apatite-like layer on their surface when soaked in simulated body fluid, being faster in the sample with a higher content of glass. The formation of the new layer began in preferential zones in both samples, depending on the different reactivity of the crystalline phases formed. A synergistic effect between HA and glass was observed, showing in the mixtures a faster bioactive behaviour than in HA and glass themselves. The obtained materials allow a good attachment, spread and proliferation of the osteoblastic-like cells and no cytotoxic effect was observed. PMID- 16701893 TI - Combining particle size distribution and isothermal calorimetry data to determine the reaction kinetics of alpha-tricalcium phosphate-water mixtures. AB - Many calcium phosphate bone substitutes are based on the use of alpha-tricalcium phosphate (alpha-TCP) powder. This compound has been intensively studied, but some aspects of alpha-TCP reactivity are still controversial. The goal of this study was to determine the setting kinetics of alpha-TCP based on a new approach that compared particle size distribution data to isothermal calorimetry data. Results indicated that alpha-TCP conversion is mostly controlled by surface reactions, with at later stages a diffusion-controlled mechanism. The presence of an X-ray amorphous alpha-TCP fraction in the crystalline alpha-TCP powder increased the dissolution rate threefold, without modifying the reaction mechanism. PMID- 16701894 TI - Interfacial reactions of glasses for biomedical application by scanning transmission electron microscopy and microanalysis. AB - Short-term physico-chemical reactions at the interface between bioactive glass particles and biological fluids are studied for three glasses with different bioactive properties; these glasses are in the SiO(2)-Na(2)O-CaO-P(2)O(5)-K(2)O Al(2)O(3)-MgO system. Our aim is to show the difference between the mechanisms of their surface reactions. The relation between the composition and the bioactive properties of these glasses is also discussed. The elemental analysis is performed at the submicrometer scale by scanning transmission electron microscopy associated with energy-dispersive X-ray spectroscopy and electron energy loss spectroscopy. After different immersion times (ranging from 0 to 96 h) of bioactive glass particles in a simulated biological solution, results show the formation of different surface layers at the glass periphery in the case of two bioactive glasses (A9 and BVA). For the third glass (BVH) we do not observe any surface layer formation or any modification of the glass composition. For the two other glasses (A9 and BVA), we observe the presence of different layers: an already observed (Si, O, Al) rich layer at the periphery, a previously demonstrated thin (Si, O) layer formed on top of the (Si, O, Al) layer and a (Ca, P) layer. We determine the different steps of the mechanisms of the surface reactions, which appear to be similar in these glasses, and compare the physico chemical reactions and kinetics using the different immersion times. The A9 glass permits the observation of all important steps of the surface reactions which lead to bioactivity. This study shows the important relationship between composition and bioactivity which can determine the medical applicability of the glass. PMID- 16701895 TI - Child maltreatment in the "children of the nineties": a cohort study of risk factors. AB - AIM: To analyze the multiple factors affecting the risk of maltreatment in young children within a comprehensive theoretical framework. METHODS: The research is based on a large UK cohort study, the Avon Longitudinal Study of Parents and Children. Out of 14,256 children participating in the study, 293 were investigated by social services for suspected maltreatment and 115 were placed on local child protection registers prior to their 6th birthday. Data on the children have been obtained from obstetric data and from a series of parental questionnaires administered during pregnancy and the first 3 years of life. Risk factors have been analyzed using an hierarchical approach to logistic regression analysis. RESULTS: In the stepwise hierarchical analysis, young parents, those with low educational achievement, and those with a past psychiatric history or a history of childhood abuse were all more likely to be investigated for maltreatment, or to have a child placed on the child protection register, with odds ratios between 1.86 and 4.96 for registration. Examining strength of effect, the highest risks were found with indicators of deprivation (3.24 for investigation and 11.02 for registration, after adjusting for parental background factors). Poor social networks increased the risk of both investigation (adjusted OR 1.93) and registration (adjusted OR 1.90). Maternal employment seemed to reduce the risk of both outcomes but adjusted odds ratios were no longer significant for registration. After adjusting for higher order confounders, single parents and reordered families were both at higher risk of registration. Reported domestic violence increased the risk of investigation and registration but this was no longer significant after adjusting for higher order variables. Low birthweight children were at higher risk of registration as were those whose parents reported few positive attributes of their babies. CONCLUSIONS: This study supports previous research in the field demonstrating that a wide range of factors in the parental background, socio-economic and family environments affect the risk of child maltreatment. By combining factors within a comprehensive ecological framework, we have demonstrated that the strongest risks are from socio-economic deprivation and from factors in the parents' own background and that parental background factors are largely, but not entirely, mediated through their impact on socio-economic factors. PMID- 16701896 TI - Cloning and characterization of a shrimp clip domain serine protease homolog (c SPH) as a cell adhesion molecule. AB - Clip domain serine protease homologs (c-SPHs) are involved in various innate immune functions in arthropods such as antimicrobial activity, cell adhesion, pattern recognition, opsonization, and regulation of the prophenoloxidase system. In the present study, we cloned a c-SPH cDNA from tiger shrimp (Penaeus monodon) hemocytes. It is 1337 bp in length with a coding region of 1068 bp consisting a protein of 355 amino acid residues. The deduced protein includes one clip domain and one catalytically inactive serine protease-like (SP-like) domain. Its molecular weight is estimated to be 38 kDa with an isoelectric point of 7.9. The predicted cutting site of the signal peptide is located between Gly(21) and Gln(22). We aligned 15 single clip domain SPH protein sequences from 12 arthropod species; the identity of these clip domains is low and that of SP-like domains is from 34% to 46%. The conserved regions are located near the amino acid residues which served as substrate interaction sites in catalytically active serine protease. Phylogenetically, the tiger shrimp c-SPH is most similar to a low molecular mass masquerade-like protein of crayfish, but less similar to c-SPHs in Chelicerata and Insecta. Nested reverse transcription polymerase chain reaction (RT-PCR) revealed that c-SPH mRNA is expressed most in tissues with the highest hemocyte abundance. Antimicrobial and opsonization activities of the molecule were not detected. The expression of c-SPH mRNA in hemocytes was up-regulated at the 12-day post beta-glucan immersion. Recombinant c-SPH could significantly enhance hemocyte adhesion. The result suggests that the shrimp c-SPH protein plays a role in innate immunity. PMID- 16701898 TI - RAG1/2 re-expression causes receptor revision in a model B cell line. AB - The expression of RAG1 and RAG2 is essential for V(D)J rearrangement of the immunoglobulin (Ig) locus in developing B cells. In mature B cells further V(D)J rearrangement is suppressed and RAG1/2 proteins decline to undetectable levels. However, there is evidence that mature B cells in the periphery may re-express RAG1/2. In humans evidence of RAG1/2 re-expression is often linked with an autoimmune state, indicating that further understanding of re-expression may be crucial to understanding immune disorders. We have investigated the molecular consequences of RAG1/2 expression in mature lymphocytes using a cell culture system (M12 and DR3). M12 (IgG+, Igkappa+ and RAG-) is a mouse B cell lymphoma. DR3 is a RAG1+/RAG2+ line derived from M12 by introduction of stable plasmids carrying RAG1 and RAG2 cDNAs. RAG1/2 mediated receptor revision occurs in the DR3 line, as evidenced by both the deletion of the endogenous rearranged Igkappa gene segment (present in the parent M12 lines) and the presence of a new Iglambda rearrangement. Gene expression profiles obtained through microarray analysis and RT-PCR found differences in expression levels between the two lines for: fibronectin, lysyl oxidase, TAP2, B220, Igkappa, TIS11B, HMG2 and DNAPKcs. Thus, the expression of RAG1/2 in a previously RAG- cell line results in multiple changes to the gene expression profile as well as receptor revision. The significance of the changes found in this model of RAG re-expression is discussed. PMID- 16701899 TI - The Vitamin E analog Trolox reduces copper toxicity in the annelid Lumbriculus variegatus but is also toxic on its own. AB - The ability of the water-soluble Vitamin E analog, Trolox, to prevent the toxic effects of copper exposure on the behavior and neuronal physiology of the freshwater oligochaete Lumbriculus variegatus was examined. Trolox produced a concentration-dependent increase in the 24 h LC(50) for copper exposure, with 100 microM Trolox elevating the LC(50) by almost seven-fold (from 0.36 to 2.43 microM). Copper exposure (0.2 microM) for 24h produced a reduction in the conduction velocity of the medial and lateral giant nerve fibers, which was prevented by 100 microM Trolox. Copper exposure (0.2 microM) for 24h also reduced the effectiveness of substrate vibration in eliciting giant nerve fiber spikes. Trolox prevented this reduction in sensory responsiveness. Trolox (100 microM) partially reversed the copper-induced (0.4 microM) decrease in touch-evoked helical swimming behavior, but had no effect on the copper-induced decrement in touch-evoked body reversal. Copper exposure (0.2 microM) for 24 h reduced the amount of spontaneous locomotion (crawling); however, Trolox did not reverse this effect. However, Trolox exposure alone produced a decrease in the distance crawled that was similar in magnitude to copper exposure. In normal worms, rapid spiking activity of the medial giant nerve fiber produces facilitation in the amplitude of the resulting muscle potentials produced by the longitudinal body wall muscles. Copper exposure had no effect on the amount of muscle potential facilitation, but Trolox exposure (100 microM) produced a significant decrease in facilitation. The results of this study indicate that many of the toxic effects of copper exposure on Lumbriculus are prevented or reduced by the antioxidant Trolox. However, the results of this study also indicate that Trolox has toxic effects on behavior and neuronal physiology. The results presented here document one of the few published reports of the detrimental effects of Vitamin E or its analogs on nervous system function or behavior. PMID- 16701900 TI - Acute cytomegalovirus infection and venous thrombosis: role of antiphospholipid antibodies. AB - Cytomegalovirus (CMV)-induced thrombosis has been reported in immunocompromised patients, such as transplant recipients and patients with AIDS. Recent cases also describe thrombotic phenomena in immunocompetent patients with CMV infection. Various mechanisms may explain the role of CMV in thrombosis: this virus can damage endothelial cells, activate coagulation factors, and induce production of antiphospholipid (aPL) antibodies. We present a case report of a previously healthy white woman with a pulmonary embolism associated with CMV infection and the presence of aPL antibodies, and we discuss the role of the aPL antibodies associated with CMV infection in the pathogenesis of thrombosis. PMID- 16701901 TI - A simple webcam-based approach for the measurement of rodent locomotion and other behavioural parameters. AB - We hereby describe a simple and inexpensive approach to evaluate the position and locomotion of rodents in an arena. The system is based on webcam registering of animal behaviour with subsequent analysis on customized software. Based on black/white differentiation, it provides rapid evaluation of animal position over a period of time, and can be used in a myriad of behavioural tasks in which locomotion, velocity or place preference are variables of interest. A brief review of the results obtained so far with this system and a discussion of other possible applications in behavioural neuroscience are also included. Such a system can be easily implemented in most laboratories and can significantly reduce the time and costs involved in behavioural analysis, especially in developing countries. PMID- 16701902 TI - Threshold and subthreshold bipolar disorders in the Sesto Fiorentino Study. AB - BACKGROUND: The DSM IV lifetime prevalence for bipolar affective disorders is reported to be between 0.4% and 1% in most surveys. DSM IV, however, fails to identify the cases clinically significant described as bipolar spectrum. The few researches that have attempted to evaluate the prevalence of these disorders in the community report lifetime prevalence figures of 5% to 10% for the whole spectrum. METHOD: The data from the Sesto Fiorentino Study, a community survey conducted by clinical interviewers using a typically clinical instrument, have been re-analysed. 2363 people out of the 2500 randomly selected in order to be representatives of the population aged 14 or more living in Sesto Fiorentino (44,000 inhabitants, Central Italy) could be evaluated according to a two-phase design. RESULTS: The lifetime prevalence rates were 0.47% for bipolar I disorder, 0.38% for bipolar II (0.85% for DSM IV bipolar disorders). A much greater number of subjects had clinically significant hypomanic symptoms that failed to fully satisfy the diagnostic criteria, totalling to 4.66% of the population. Compared with depressives without any sign of bipolarity (N=297), these "subthreshold bipolar" cases had significantly greater comorbidity for Generalised Anxiety Disorder, Obsessive-Compulsive Disorders and Anorexia Nervosa, as well as with Obsessive-Compulsive Passive-Aggressive, Paranoid, Borderline personality disorders. Their depressive symptom pattern cases differed from that of the "pure" unipolar cases for a significantly greater frequency of psychic agitation, psychotic symptoms and hyperphagia. LIMITATIONS AND CONCLUSIONS: The main limitation of the study is its retrospective nature, whereas it confirms the clinical relevance of even softer forms of bipolarity on epidemiological grounds. PMID- 16701903 TI - Hippocampal volume in chronic posttraumatic stress disorder (PTSD): MRI study using two different evaluation methods. AB - The hippocampus is discussed as one of the key regions in the pathogenesis of Posttraumatic Stress Disorder (PTSD). MRI results concerning the volume of the hippocampus are, however, inconsistent. This may be due to the heterogeneity of patients' traumata or postprocessing of the imaging data. To overcome these problems, the present study investigates volume changes in well-characterized chronic PTSD patients in comparison to controls using two different evaluation methods. MATERIAL AND METHODS: 15 patients with chronic PTSD, traumatized at the same air show plane crash in 1988 (Ramstein, Germany), and 15 matched healthy controls participated in this study. All patients suffered from significant impairment by the PTSD; none had a history of drug or alcohol abuse. Hippocampus volume changes were processed by a semi-automated standard procedure performed with BRAINS2 as well as the voxel based morphometry (VBM) using SPM2. RESULTS: No differences in total brain grey or white matter were detected between patients and controls. No differences in total hippocampal volume or in right and left parts were seen, even when hippocampal volumes were corrected by total brain volume or correlated with clinical data. Finally, no significant differences were detected between patients and controls in hippocampal regions using VBM. DISCUSSION: This is the first study examining long-term changes in hippocampal volumes in chronic PTSD patients compared to matched controls using two different evaluation methods. Neither conventional volumetry nor VBM could detect any differences in the volume and structure. This supports the hypothesis that previously described hippocampal volume reduction is not necessarily due to PTSD or at least that, after 15 years, volume changes have been restored or have not yet developed. PMID- 16701904 TI - Expression of TOLL-like receptors (TLR) by bovine antigen-presenting cells potential role in pathogen discrimination? AB - Invading pathogens are controlled by the innate and adaptive arms of the immune system. Adaptive immunity, mediated by B and T lymphocytes, recognises pathogens via high affinity receptors. However, the establishment of a primary adaptive immune response is not rapid enough to eradicate invading microorganisms as it involves cell proliferation, gene activation and protein synthesis. More rapid defence mechanisms are provided by innate immunity, which recognises invading pathogens by germ-line-encoded pattern recognition receptors. Recent evidence shows that this recognition can mainly be attributed to the family of TOLL-like receptors (TLR). Binding of pathogen-associated molecular patterns to TLR induces the production of reactive oxygen and nitrogen intermediates, pro-inflammatory cytokines, and up-regulates expression of co-stimulatory molecules, subsequently initiating the adaptive immunity. In this paper, we will discuss the current knowledge with regards to the TLR, and in particular the bovine family of TLR. In addition, we will show the expression of TLR mRNA in bovine antigen-presenting cell subsets, summarise the discovery and the critical roles of TLR2 in host defence against Mycobacteria, and provide evidence for a mycobacteria species specific response of bovine macrophages. PMID- 16701906 TI - Megaprimer-mediated domain swapping for construction of chimeric viruses. AB - Clones that encode viral genomes constructed from two viruses with contrasting biological properties have been widely used in studies of viral-host interactions, particularly when the objective is to determine the identity of the viral component recognized by the host in a resistant response, known as the avirulence factor. This paper presents an efficient method based on megaprimer mediated domain swapping for the construction of clones encoding chimeric viral genomes as a versatile and widely applicable alternative to conventional restriction enzyme digestion and ligation methods. Potato virus X (PVX)-derived vectors expressing genes encoding fluorescent proteins were used to demonstrate this concept. The cyan fluorescent protein (CFP) gene was cloned into a binary PVX vector and subsequently replaced with the yellow fluorescent protein (YFP) gene using the megaprimer amplification reaction. DNA fragments up to 1480 bp could be replaced efficiently and quickly. Most viral clones showed the expected change in phenotype without altered infectivity. Sequence analysis revealed mutations were not introduced into the four domain-swapped plasmids. This approach will provide a valuable tool for determining which domains of a viral genome are essential for infectivity, avirulence, or otherwise determine biologically significant properties of plant viruses. PMID- 16701905 TI - B subunit of E. coli enterotoxin as adjuvant and carrier in oral and skin vaccination. AB - Mucosal sites are one of the main natural ports of entry into the body. Stimulation of a local response by antibodies as the systemic protection may enhance the efficacy of non-living vaccines, and allow for vaccination by subunit vaccines without the need for injection. Mucosal or skin vaccination necessitates a suitable adjuvant and carrier. Escherichia coli heat-labile enterotoxin (LT) and its B subunit (LTB) have been found to be effective adjuvants. The aim of this study was to efficiently produce and purify recombinant LTB (brLTB), and examine its adjuvant and carrier properties. The gene encoding LTB was cloned and expressed in E. coli, and the product was found to have a pentameric form with the ability to bind the cell receptor, GM1 ganglioside. A one-step method for efficient purification and concentration of brLTB was developed. Both oral and intramuscular vaccination with purified brLTB yielded high antibody titers, which detected the whole toxin. In an attempt to test its adjuvant characteristics, brLTB was mixed with either BSA or a recombinant protein (rKnob of egg drop syndrome adenovirus) and delivered intramuscularly, orally or transcutaneously. The addition of brLTB significantly elevated the antibody response in groups vaccinated orally and transcutaneously, but had no influence in injected groups. Vaccination with another recombinant protein, (viral protein 2 of infectious bursal disease virus) supplemented with brLTB did not elevate the antibody response, as compared to vaccination with the antigen alone. These results demonstrate that the addition of brLTB makes oral and transcutaneous vaccination with protein antigens possible. PMID- 16701908 TI - Effects of manganese on chemically induced food search behaviour of the Norway lobster, Nephrops norvegicus (L.). AB - The decapod Norway lobster, Nephrops norvegicus (L.), lives on muddy sediments rich in manganese (Mn). In hypoxic conditions, manganese is reduced and released from the sediment, so increased concentrations of dissolved Mn(2+) become bioavailable. In excess, manganese acts as a neurotoxin and may inhibit vital functions of benthic organisms, such as muscle contraction. We investigated in a laboratory flume experiment, the effect of environmentally realistic concentrations of manganese (0.1 and 0.2mM for 12 days) on the food search behaviour of N. norvegicus. We found that lobsters exposed to manganese had a more than doubled reaction time to food odour stimuli compared to the controls (p<0.05). In addition, manganese exposure reduced the number of N. norvegicus reaching the food stimuli source. Compared to the controls where 86% reached the stimuli source, only about half of the lobsters exposed to 0.1mM Mn and one-third of the lobsters exposed to 0.2mM Mn reached the stimuli source (p<0.05 and 0.001, respectively). There was no significant difference between treatments in the number of lobsters leaving their shelter or in the time from reaction until leaving the shelter and there was no difference in search time for those animals that eventually did locate the stimuli source. This study shows that environmentally realistic manganese concentrations affect parts of the food search behaviour of N. norvegicus, likely due to impaired chemosensory ability or reduced motivation for feeding. Thus, the ability of N. norvegicus to detect and find food can be reduced in areas with high manganese concentrations, with possible consequences on individual and population levels. PMID- 16701909 TI - Roles of cardiac ryanodine receptor in heart failure and sudden cardiac death. AB - Calcium (Ca2+) plays an important role as a messenger in the excitation contraction coupling process of the myocardium. It is stored in the sarcoplasmic reticulum (SR) and released via a calcium release channel called the ryanodine receptor. Cardiac ryanodine receptor (RyR2) controls Ca2+ release, which is essential for cardiac contractility. There are several molecules which bind and regulate the function of RyR2 including calstabin2, calmodulin, protein kinase A (PKA), phosphatase, sorcin and calsequestrin. Alteration of RyR2 and associated molecules can cause functional and/or structural changes of the heart, leading to heart failure and sudden cardiac death. In this review, the alteration of RyR2 and its regulatory proteins, and its roles in heart failure and sudden cardiac death, are discussed. Evidence of a possible novel therapy targeting RyR2 and its associated regulatory proteins, currently proposed by investigators, is also included in this article. PMID- 16701911 TI - The correlation between D90 and outcome for I-125 seed implant monotherapy for localised prostate cancer. AB - BACKGROUND AND PURPOSE: In 1998 Stock and Stone demonstrated a dose response relationship correlating D90 with probability of biochemical control and showed that a D90 of 140 Gy is a highly significant factor in predicting PSA relapse free survival (PSA-RFS). Although, a mean D90 of over 140 Gy was achieved in our series, there is nevertheless a normal distribution with 20% of patients achieving a D90 of less than 120 Gy. We have analysed the possible causes for the low D90 and the impact on outcome. PATIENTS AND METHODS: Prospective data from 667 patients treated between 1995 and 2001 by I-125 seeds prostate implant as monotherapy were analysed. Post-implant dosimetry was performed on 413 patients. D90 and other indices were calculated for each patient. Statistical analysis was performed on D90 dose to identify the correlation that would predict the 8.2 years PSA relapse free survival as defined by the American Society for Therapeutic Radiology and Oncology (ASTRO). RESULTS: Correlation between D90 and outcome shows no significant difference for the whole population between those who receive greater or less than 140 Gy (P=0.43) and there was also no difference for those receiving more or less than 130 Gy (P=0.14). Subgroup analysis by risk group, however, showed that for low risk patients there was a significant correlation between D90 and PSA control (P<0.01). Although, post-implant dosimetry was performed 6-8 weeks after brachytherapy, post-implant CT still showed variable levels of oedema compared with the pre-implant ultrasound. A statistically significant relationship was shown between D90 and the ratio between CT and ultrasound volume (P<0.01) which suggests that some low D90s may be related to persistent oedema at the time of calculation. Segmental analysis of a subgroup of 32 patients showed that the dose was most often deficient in the anterior basal segment of the gland. CONCLUSIONS: D90 was found to be a good discriminator for those with low risk where failure to achieve local control is likely to be the dominant cause of PSA failure. No significant dose response relationship between D90 and PSA was found in the intermediate and high-risk population of patients. This could be due to (1) the presence of oedema or discrepancy between pre- and post-implant volumes causing a low D90, (2) the possibility that the underdosed area could be situated where there is unlikely to be tumour, (3) the fact that biochemical control does not equate to local control because some patients fail outside the prostate, particularly in the high and intermediate risk patients, (4) if D90 is a good discriminator only for low risk patients, the absence of a dose response correlation in this series which contained 53.8% intermediate and high risk patients could be related to case mix. PMID- 16701910 TI - Quantification of homogentisate-1,2-dioxygenase expression in a fungus degrading ethylbenzene. AB - A quantitative real time reverse transcription polymerase chain reaction (qRT PCR) assay was utilized to quantify the expression of ElHDO in the fungus Exophiala lecanii-corni during the biodegradation of ethylbenzene and other volatile organic pollutants. The assay was applied to measure the impact of pollutant mixtures on ElHDO expression relative to that of a housekeeping gene (18S rRNA). Three compounds were tested in mixtures with ethylbenzene: methyl propyl ketone, phenylacetate and o-xylene. These chemicals repressed, induced, or had no effect on ethylbenzene degradation, respectively. The results demonstrate that the gene target expression value (T(N)) is a useful parameter for evaluating the effect of pollutant mixtures on gene expression. T(N) was found to reflect macroscopic changes in ethylbenzene utilization rates although these two parameters were not related in a linear fashion for all compounds. The assay was log-linear over 5 orders of magnitude of RNA concentration and reproducible between samples (the largest T(N) standard deviation was 20%). The comparative qRT-PCR assay used in this research represents a viable alternative to absolute quantification methods to monitor in situ fungal gene expression in natural and engineered environmental systems. PMID- 16701912 TI - Aspergillus spp., distribution, population composition and ochratoxin A production in wine producing vineyards in Greece. AB - Vineyard surveys of 11 wine producing grape cultivars, were carried out in sixteen vineyards, in five winemaking regions in Greece, during 2002 and 2003. The occurrence of various Aspergillus spp. in bunches of berries at setting, veraison and ripening at harvest time was investigated. Aspergillus niger aggregate and A. carbonarius were predominantly isolated from sampled berries. Although the prevailing Aspergillus spp. isolates belonged to A. niger aggregate, isolates of A. carbonarius were the most efficient ochratoxin A (OTA) producers. Of 50 tested isolates of A. carbonarius 42% produced amounts of OTA, exceeding 25 ppb, while none of the 85 isolates of A. niger aggregate tested produced above 16 ppb. PMID- 16701913 TI - Characterisation of lactic acid bacteria isolated from fermented milk "laban". AB - The technological properties of 96 lactic acid bacteria isolated from Lebanese traditional fermented milk "laban" were characterised. They were classified by phenotypic and biochemical analyses as Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus, thus indicating that laban is a fermented milk similar to yogurt. Most strains of L. bulgaricus (87.5%) exhibited a high acidification activity, whereas strains of streptococci showed low acidification ability. 33.3% of streptococci strains and 25% of lactobacilli strains displayed similar acidification performances as European strains. Results obtained for syneresis, texture and rheological parameters led us to consider that isolated strains were not low polymer-producing strains. Some of them displayed interesting characteristics such as low syneresis and high values for rheological parameters. The major flavour compounds found in pure cultures were acetaldehyde, acetone, 2-butanone, dimethyl disulfide, acetoin, 2,3-butanedione, 2,3-pentanedione, and acetic, hexanoic and butanoic acids. Acetaldehyde (7.4%) and organic acids (48.3%) were mainly produced by L. bulgaricus strains, whereas streptococci cultures contained high relative levels of 2,3-butanedione and acetoin, which represented around 82% of the total flavour compounds. Finally, strains isolated from laban samples exhibited different technological properties than those used in yogurt production, thus conferring specific characteristics to this product. PMID- 16701907 TI - Large-scale amplification, cloning and sequencing of near full-length HIV-1 subtype C genomes. AB - Full-length HIV-1 genome sequencing provides important data needed to address several vaccine design, molecular epidemiologic and pathogenesis questions. A protocol is presented for obtaining near full-length genomes (NFLGs) from subjects infected with HIV-1 subtype C. This protocol was used to amplify NFLGs from 244 of 366 (67%) samples collected at two clinics in Durban, South Africa (SK and PS). Viral load was directly associated with frequency of successful NFLG amplification for both cohorts (PS; p = 0.005 and SK; p < 0.001). Seventeen of 38 initially NFLG-negative SK samples had variation within the PCR primer binding sites, however only 3 of these were successfully re-amplified using re-designed primers homologous to the target viruses. NFLGs were obtained from 7 of 24 PBMC samples processed from subjects whose plasma did not yield a NFLG. Stable plasmid clones were obtained from all 244 NFLG-positive PCR products, and both strands of each genome were sequenced, using a primary set of 46 primers. These methods thus allow the large-scale collection of HIV-1 NFLGs from populations infected primarily with subtype C. The methods are readily adaptable to other HIV-1 subtypes, and provide materials for viral functional analyses and population based molecular epidemiology studies that include analysis of viral genome chimerization. PMID- 16701914 TI - Molecular epidemiology of rabies virus strains isolated from wild canids in Northeastern Brazil. AB - Rabies in wild canids in Northeastern Brazil is frequent and has been reported for some time, with episodes of rabies transmission from these animals to humans also reported. In this study, we analyzed the antigenic and genetic profiles of the rabies virus nucleoprotein gene, isolated from 20 samples taken from domestic animals and wild canids located in the Northeastern region of Brazil. All viruses isolated from domestic animals (dogs and cats) belonged to the antigenic variant 2 (AgV2). Among the wild animal samples, only four were AgV2, and nine showed a divergent antigenic profile. Phylogenetic analysis revealed two Brazilian clusters. Cluster 1 (Brazilian domestic carnivore-related strains) showed two subclusters, called 1A and 1B, and cluster 2 (Brazilian wild canid-related strains) also showed two subclusters, called 2A and 2B. The majority of the samples with divergent antigenic strains segregated into subcluster 2A. The intracluster identity of cluster 1 was 95.6% and that of cluster 2, 92.4%. When clusters 1 and 2 were compared, an identity of 88.6% was found. The genetic analysis of wild canid samples performed in this study indicates that there are two distinct rabies cycles among canids in Brazil, one represented by domestic canids and the other by wild canids. This study shows that the virus samples isolated in Northeastern Brazil are region and species-specific. PMID- 16701915 TI - Polyion complex micelles for photodynamic therapy: incorporation of dendritic photosensitizer excitable at long wavelength relevant to improved tissue penetrating property. AB - A polymeric micelle (DPcZn/m) system, which is formed via an electrostatic interaction of anionic dendrimer phthalocyanine (DPcZn) and poly(ethylene glycol) poly(l-lysine) block copolymers (PEG-b-PLL), was prepared for use as an effective photosensitizer for photodynamic therapy. DPcZn/m exhibited strong Q band absorption around 650 nm, a useful wavelength for high tissue penetration. Dynamic light scattering studies indicated that the DPcZn/m system has a relevant size of 50 nm for intravenous administration. Under light irradiation, either DPcZn or DPcZn/m exhibited efficient consumption of dissolved oxygen in a medium to generate reactive oxygen species and an irradiation-time-dependent increase in photocytotoxicity. The photodynamic efficacy of the DPcZn was drastically improved by the incorporation into the polymeric micelles, typically exhibiting more than two orders of magnitude higher photocytotoxicity compared with the free DPcZn at 60-min photoirradiation. PMID- 16701917 TI - Treatment-related stresses and anxiety-depressive symptoms among Chinese outpatients with type 2 diabetes mellitus in Hong Kong. AB - Based on focus group findings, a descriptive instrument was developed to examine the relationship among treatment-related stress, anxiety and depressive symptoms, distress, and impairment of 333 Chinese outpatients with type 2 diabetes mellitus (DM) in Hong Kong. It was found that the main stresses included fears of diabetes complications, work impairment, lifestyle adjustment, stigmatization, and discrimination. Over 1/4 of patients concealed their DM from family members in order not to make the latter worry. 28.3% felt that life was not worth living. 33.6% of patients exhibited four or more anxiety and depressive symptoms accompanied by significant distress and/or impairment. These patients were more likely to be female, of lower educational level, and unemployed. Diabetes complications, concealment of DM, and feeling of being a burden to the family predicted anxious-depressed status. The study showed that treatment-related stresses and anxiety-depressive symptoms were common and associated among Chinese diabetes outpatients in Hong Kong. PMID- 16701919 TI - Relationships between family conflict, perceived maternal verbal messages, and daughters' disturbed eating symptomatology. AB - The objective was to investigate relationships between family conflict and the perceived communication of negative messages regarding weight and shape from mothers to daughters, and daughters' disordered eating symptomatology. A correlational study was conducted in which a questionnaire was completed by 315 women aged between 14 and 28 years. The sample comprised both secondary school students (n=196) and university students (n=119). Disordered eating symptomatology was operationalised as drive for thinness, body dissatisfaction, and bulimic symptoms. Family conflict and the perceived frequency of negative messages regarding weight and shape communicated from mothers were also reported by participants. With current BMI treated as a covariate, support was provided for the proposition that frequency of negative messages is a more important contributor to disordered eating symptomatology than family conflict for both secondary school students and university students. However, for secondary students there was no relationship between family conflict and drive for thinness. Recommendations for future research are provided, including the possibility of considering the perception of both positive and negative weight related messages, and the relative importance of such messages when conveyed by people other than the maternal figure, such as fathers, siblings and peers. PMID- 16701918 TI - C(-106)T polymorphism of the aldose reductase gene and the progression rate of diabetic retinopathy. AB - PURPOSE: To study the C(-106)T polymorphism in the promoter of the aldose reductase (ALR2) gene: (a) its local prevalence and (b) its modulation of the susceptibility for developing retinopathy. METHODS: DNAs of 96 control subjects and 53 long-standing (duration 17.9+/-5.4 years) type-2 diabetic patients were analyzed by PCR-RFLP with BfaI enzyme. Retinopathy was graded with 2-eye, 7-field fundus color photography. The IMF-HbA1c was the arithmetic mean of all HbA1c's of each patient. RESULTS: The genotypes in the controls were CC=57 (59.4%), CT=32 (33.3%) and TT=7 (7.3%), with Hardy-Weinberg chi(2)=0.793 (p>0.50). Among 53 diabetics, CC=24 (45.3%), CT=26 (49.0%) and TT=3 (5.7%). The correlation between IMF-HbA1c and retinopathy progression rate was significant on CC (r=0.6102, p=0.0072) but not in CT+TT genotypes (r=0.26, p=0.1811). CONCLUSIONS: In Chilean adults, the frequency of the C(-106)T polymorphism of the ALR2 gene was similar to that reported by others. Type-2 diabetics with the CC genotype were more susceptible for developing retinopathy as a result of chronic hyperglycemia than those with the CT or TT genotype. PMID- 16701920 TI - Effects of ApoE genotype and mild cognitive impairment on implicit learning. AB - The goals were to investigate implicit learning in mild cognitive impairment (MCI), and to determine the relations of implicit learning systems to apolipoprotein E (ApoE) genotype in healthy controls. Elderly controls grouped by ApoE status (ApoE-e4 allele carriers versus ApoE-e4 allele non-carriers) and MCI patients participated in the study. Individuals in all three groups completed both contextual cueing and serial reaction time (SRT) tasks. In the former, people learn to use repeated spatial configurations to facilitate search for a target, whereas in the latter, they learn to use subtle sequence regularities to respond more quickly and accurately to a series of events. Results revealed that healthy elderly individuals carrying the ApoE-e4 allele showed contextual cueing deficits compared to those who did not carry the ApoE-e4 allele. Further, elderly controls carrying the ApoE-e4 allele revealed similar amounts of contextual cueing as the MCI group, while the non-carriers performed better. Sequence learning, by contrast, was uninfluenced by either MCI or by ApoE genotype in healthy controls. This study provides further support for the medial temporal lobe dysfunction and relative integrity of fronto-striatal systems in MCI, and indicates the influence of ApoE genotype on implicit learning even in healthy older individuals without cognitive impairment. PMID- 16701921 TI - Nutritional routines and attitudes among doctors and nurses in Scandinavia: a questionnaire based survey. AB - BACKGROUND AND AIMS: Hospital malnutrition is prevalent, but nutritional practice in hospitals has a low priority. To improve the quality in nutritional routine, ESPEN has developed standards to improve the inadequate and insufficient nutritional treatments seen today. However, there is a discrepancy between the standards and clinical practice. This study was conducted to investigate nutritional practice in different hospital settings in relation to these standards (e.g.: screening of all patients, assessment of at-risk patients) among Scandinavian doctors and nurses. METHODS: A questionnaire about nutritional attitudes and routine was mailed to doctors and nurses in Denmark, Sweden and Norway. RESULTS: Altogether, 4512 (1753 doctors, 2759 nurses) answered the questionnaire. Both screening and assessment of at-risk patients differ between the countries. Nutritional screening was more common in Denmark (40%), compared to Sweden (21%) and Norway (16%). Measuring dietary intake in nutritional at-risk patients was more common in Denmark (46%), compared to Sweden (37%) and Norway (22%). However, all countries agreed that nutritional screening (92%, 88%, 88%) and measuring dietary intake (97%, 95%, 97%) were important, Denmark, Sweden and Norway, respectively. CONCLUSION: There is a large discrepancy between nutritional attitudes and practice. The standards suggested from the ESPEN are not fulfilled. PMID- 16701922 TI - Effects of milk enriched with omega-3 fatty acid, oleic acid and folic acid in patients with metabolic syndrome. AB - BACKGROUND & AIMS: Patients with metabolic syndrome (MS) have increased cardiovascular risk factors. Dietary modifications mainly polyunsatturated fatty acids intake, can improve them. The present study was performed to assess the effects of enriched milk with omega-3 and oleic fatty acids, folic acid and vitamin E, in these patients. METHODS: We performed a randomized, placebo controlled and open clinical trial, among 72 patients with MS for 3 months. Thirty-six of them consumed 500 cm(3) per day of semi-skimmed milk (control group), and the others consumed 500 cm(3) per day of enriched milk (test group). Daily supplements in this group were 5.7 g of oleic acid, 0.2g of omega-3 fatty acid, 150 microg of folic acid and 7.5mg of vitamin E. Serum for total and HDL cholesterol, triglycerol, Apo B, glucose, insulin, hs-CRP, homocysteine and fatty acids contents in serum phospholipids, was obtained at the beginning and at the end of the study. LDL cholesterol was calculated by Friedewald formula. RESULTS: Four patients in the test group, and two in the control group dropped out. In the test group a decrease in serum total cholesterol (-6.2%, P=0.006), LDL cholesterol (-7.5%, P=0.032), triglycerol (-13.3%, P=0.016), Apo B (-5.7%, P=0.036), glucose (-5.3%, P=0.013), and homocysteine (-9.5%, P=0.00) was observed. Any of these parameters changed in the control group. CONCLUSIONS: Dietary supplementation with 500 cm(3) of enriched milk with omega-3 fatty acid, oleic acid and folic acid, reduces serum tryglicerides, total and LDL cholesterol, Apo B, glucose and homocysteine in patients with MS. This milk is well tolerated and accepted by the patients. PMID- 16701923 TI - Effects of creatine supplementation on nutritional status, muscle function and quality of life in patients with colorectal cancer--a double blind randomised controlled trial. AB - BACKGROUND & AIM: Nutritional status frequently deteriorates during chemotherapy in cancer. This is associated with a poor outcome. Since creatine supplementation has shown promising results in various diseases, we investigated the effects of creatine on nutritional status in patients with colorectal cancer undergoing chemotherapy. METHODS: Thirty patients with colorectal cancer undergoing chemotherapy were randomised to receive either creatine (n=16) or placebo (n=15) for 8 weeks. Body composition was determined with bioelectrical impedance analysis; muscle function by hand grip, hip flexion and knee extension strength and quality of life (QoL) was assessed by the QLQ30 questionnaire. RESULTS: Neither muscle function, body cell mass (BCM) nor QoL improved, but phase angle, a marker of BCM and cell integrity, increased significantly in the intervention patients (5.3+/-0.3 degrees to 5.4+/-0.2 degrees , P=0.030). Evaluating patients with different chemotherapy regimens, however, only intervention patients undergoing less aggressive chemotherapy were shown to benefit, increasing phase angle (5.11+/-0.22 degrees to 5.51+/-0.30 degrees , P=0.043) as well as BCM (27.22+/-2.85 to 29.60+/-3.54kg, P=0.043). CONCLUSION: Creatine failed to improve muscle mass or function and QoL in colorectal cancer patients but improved bioimpedance parameters that are predictive of poor outcome. Creatine might therefore be useful in patients with milder chemotherapy in order to maintain or increase BCM whereas patients undergoing aggressive chemotherapy however are not likely to benefit. PMID- 16701924 TI - Influence of genetic and environmental factors on the immunogenicity of Hib vaccine in Gambian twins. AB - The differences in incidence rates of Haemophilus influenzae type b disease and the variation in Hib conjugate vaccine efficacy achieved among different ethnic groups suggest genetic influences on the immune response to Hib vaccine. The serum anti-PRP antibody concentration of 43 monozygotic (MZ) and 147 dizygotic (DZ) twin pairs in the Gambia was measured using a standardised Hib ELISA. Intrapair correlations for MZ and DZ twin pairs were compared and heritability in antibody responses to Hib conjugate vaccine was estimated to be 51% (95% CI: 32 66%), indicating a significant genetic contribution in the response. We conclude that genetic factors may be involved in the variation in immune response to Hib vaccine observed in different populations and may contribute to cases of vaccine failure. PMID- 16701925 TI - Safety and immunogenicity of Towne cytomegalovirus vaccine with or without adjuvant recombinant interleukin-12. AB - The Towne, human cytomegalovirus (CMV) vaccine is safe and immunogenic but has not prevented infection at doses tested to date. We administered 3000 pfu Towne CMV vaccine, with or without adjuvant recombinant interleukin-12 (rhIL-12), to CMV-seronegative healthy volunteers and then measured CMV gB-specific IgG titers and CMV-specific CD4+ and CD8+ T cell proliferation and IFNgamma expression after stimulation with whole viral lysate and immunodominant peptide CMV antigens. Adjuvant rhIL-12 at doses up to 2 microg were well-tolerated and associated with (1) dose-related increases in peak anti-CMV gB IgG titers (though not in sustained titers), (2) dose-related increases in the weak CMV viral lysate specific CD4+ T cell proliferation responses induced by vaccine alone after 360 days of follow-up, and (3) decreases in the very robust CMV IE-specific peak CD4+ T cell and Day 360 CD8+ T cell proliferation responses induced by the vaccine alone. Also, qualitative CD8+ T cell IFNgamma responses to stimulation with the immunodominant CMV antigen, pp65, tended to occur more frequently in vaccinees who received 0.5-2.0 microg rhIL-12 compared to lower dose or no rhIL-12. Thus, adjuvant IL-12 may be a promising strategy for improving antibody and T cell immune responses to a CMV vaccine. PMID- 16701926 TI - Changes in scaphoid bone density after acute fracture. AB - The radiographic density of the proximal fragments of 16 scaphoid fractures was assessed on scaphoid series radiographs taken at 6 to 12 weeks. In addition, dual energy X-ray absorptiometry measurements of bone mineral density in the distal radius and proximal and distal fracture fragments were performed at 1 to 2 weeks and 6 to 12 weeks. Median reductions of 9% and 10% were observed in bone mineral density in the proximal fracture fragment and the distal radius respectively, but these did not correlate with the radiographic density of the proximal fragment. A greater median reduction in bone mineral density (27%) was observed in the distal fracture fragment and more bone loss occurred at this site when there was an apparent increase in the radiographic density of the proximal fragment (median fall of 0.23 g/cm2 versus 0.14 g/cm2). Thus, apparent increased radiographic density of the proximal fragment may be due to increased bone loss from the distal fragment. PMID- 16701927 TI - Assessments of different kinds of stems by experiments and FEM analysis: appropriate stress distribution on a hip prosthesis. AB - BACKGROUND: It is now recognized that initial stability is essential for avoidance of thigh pain in hip replacement. The initial stability corresponds to an optimal stress distribution of cementless orthopedic implants. Although the relationship between the contour and stress at the fixation site has been analyzed, guidelines on stem design have not been established. METHODS: Finite element models of three currently-used stems were constructed for a computer simulation. Contact stress at the fixation site of a joint prosthesis was analyzed by an explicit three-dimensional finite element method. The stress immediately after applying load using a film or sensor which can measure contact stress was observed. The situation of the initial fixation about the specific part which becomes important clinically based on the results was clarified. FINDINGS: We introduced fluctuation area as a measure to evaluate the primary fixation of femoral stems. It was found that the stress distribution on the PerFix SV stem fluctuated with a slight disturbance. On the Intra-Medullary Cruciate stem, the high stress areas were distributed on the proximal area and under the pin. The high stress area on the VerSys stem were spread on the medial side. INTERPRETATION: This study highlights the mechanical instability of the fixation site of joint prostheses, and thus suggests that the general idea that unconditionally recommends a larger area for the fixation site of joint prostheses should be revised. PMID- 16701929 TI - The impact of UV-B and sulphur- or copper-containing solutions in acidic conditions on chlorophyll fluorescence in selected Ramalina species. AB - Ramalina maciformis and Ramalina lacera were exposed to different solutions and UV-B to seek for alterations in the PSII photosynthetic quantum yield (F(v)/F(m)), in response to chemicals and radiation. For R. maciformis, significant alterations of the F(v)/F(m) ratio occurred only in response to different bisulphite solutions. The F(v)/F(m) ratio decreased most in R. maciformis and R. lacera following exposure to 5 and 1 mM bisulphite, respectively. Significant differences in F(v)/F(m) ratios were observed for R. lacera in response to different solutions and light at different wavelengths, this being synergistic. The PSII system was unaffected by simulated acid rain in both lichens. R. maciformis, in particular, may survive limited acid rain exposure owing to high Ca oxalate accumulation. The F(v)/F(m) ratio decreased most in R. lacera following short-term exposures to CuSO(4), suggesting that this species is more sensitive to Cu ions under acidic conditions. PMID- 16701928 TI - Measures of ozone concentrations using passive sampling in forests of South Western Europe. AB - Ambient ozone concentrations were measured with passive samplers in the framework of the EU and UN/ECE Level II forest monitoring programme. Data from France, Italy, Luxembourg, Spain and Switzerland are reported for 2000-2002, covering the period from April to September. The number of plots increased from 67 in 2000 to 83 in 2002. The year 2001 experienced the highest ozone concentrations, reflecting more stable summer meteorological conditions. Average 6-month ozone concentrations above 45 ppb were measured this year in 40.3% of the plots, in contrast with the less than 21% measured in the other 2 years. Gradients of increasing ozone levels were observed from North to South and with altitude. Comments are made on the regional trends and on the time frame of the higher ozone episodes. Also, some recommendations enabling a better comparison between plots are provided. PMID- 16701930 TI - Phytotoxicity data safeguard the performance of the recipient plants in leachate irrigation. AB - Leachates from an operating and a closed landfill were examined for their phytotoxicity by seed germination/root elongation tests using seeds of Brassica chinensis and Lolium perenne. Their EC50s ranged from 3% to 46% v/v, which varied remarkably with the operating status of the landfills. Seedlings of twelve tree species were grown in pots, which were irrigated with landfill leachate at the EC50 levels, with tap water as control. No tree mortality or growth inhibition was observed after 90 days of leachate application. Chlorophyll fluorescence measurement also showed that plants receiving leachate did not suffer from a decline in photosynthetic efficiency. Litsea glutinosa and Hibiscus tiliaceus had remarkable growth, and other non-N-fixers were not inferior to the N-fixing Acacia auriculiformis. Leachate irrigation improved soil N content, though P deficiency is still a problem. The seed bioassay provided a conservative estimate of the phytotoxicity of landfill leachate. Plants irrigated can be protected from growth inhibition when the leachate irrigation plan is designed with reference to phytotoxicity data. PMID- 16701931 TI - Review of the toxicity of chemical mixtures containing at least one organochlorine. AB - An analysis of current research on mixture toxicity was conducted by critically reviewing published journal articles. The scope was limited to complex mixtures (more than two components) where at least one component was a chlorinated organic chemical. Although the basics of dose-response are widely accepted for mixtures, a number of technical issues, including dose metrics and the unquantified influence of toxicity modifying factors, confound data interpretation and restrict the ability to establish reliable determinations of the presence, nature, and extent of additivity. Lack of knowledge about dose level influences and species-specific variations contribute further interpretational limitations. Within this context, available data indicates that most tested mixtures are near or below simple dose/concentration additivity. Exceptions (both positive and negative) tend to occur when tested mixtures have only a few components or where sensitive whole organism or sub-organismal changes are used as the response metric. Available information does not routinely identify the presence of chlorine as a marker either of a particular type of toxicity or consistently greater potency. The most profound difficulty is the problem of clearly defining when and why similarity and dissimilarity of toxic action is expected for a particular mixture. This impediment largely results from the lack of a generally accepted, technical classification for mode/mechanism of toxic action coupled with the lack of a generally accepted classification scheme for mode/mechanism of toxicity interactions. PMID- 16701932 TI - The development and regulation of occupational exposure limits in Taiwan. AB - The occupational exposure limits (OELs) in Taiwan was promulgated in 1974 and has been revised five times since then. Many of the OELs were adopted from the most recent ACGIH TLVs and US OSHA PELs. A total of 483 chemicals were listed in the current Taiwan OELs Standard. The procedures of OELs development in Taiwan include the IOSH organized a recommended exposure limits (RELs) Committee to select the target chemicals and to recommend the RELs through literature review based on the health effects in the first stage, then, the CLA put policy needs, economical and technical feasibility into consideration and set up the final OELs at the second stage. A standard operation manual of RELs Committee has been developed. Based on our experience, several issues including the participation of representatives from a comprehensive spectrum, communication/education and training/enforcement, continuous collection of the local exposure data and health hazard information, use of health risk assessment, consideration of economic, and technical feasibility, as well as the globalization and information and experience sharing are critical in developing the appropriate OELs. Three examples including benzene, crystalline silica, and 2-methoxy ethanol are given to demonstrate the operation of system. PMID- 16701933 TI - Review of the toxicity of chemical mixtures: Theory, policy, and regulatory practice. AB - An analysis of current mixture theory, policy, and practice was conducted by examining standard reference texts, regulatory guidance documents, and journal articles. Although this literature contains useful theoretical concepts, clear definitions of most terminology, and well developed protocols for study design and statistical analysis, no general theoretical basis for the mechanisms and interactions of mixture toxicity could be discerned. There is also a poor understanding of the relationship between exposure-based and internal received dose metrics. This confounds data interpretation and limits reliable determinations of the nature and extent of additivity. The absence of any generally accepted classification scheme for either modes/mechanisms of toxic action or of mechanisms of toxicity interactions is problematic as it produces a cycle in which research and policy are interdependent and mutually limiting. Current regulatory guidance depends heavily on determination of toxicological similarity concluded from the presence of a few prominent constituents, assumed from a common toxicological effect, or presumed from an alleged similar toxic mode/mechanism. Additivity, or the lack of it, is largely based on extrapolation of existing knowledge for single chemicals in this context. Thus, regulatory risk assessment protocols lack authoritative theoretical underpinnings, creating substantial uncertainty. Development of comprehensive classification schemes for modes/mechanisms of toxic action and mechanisms of interaction is needed to ensure a sound theoretical foundation for mixture-related regulatory activity and provide a firm basis for iterative hypothesis development and experimental testing. PMID- 16701934 TI - Stability of cytochromes P450 and phase II conjugation systems in precision-cut rat lung slices cultured up to 72 h. AB - The objective of the present study was to evaluate the stability of cytochrome P450 enzymes and of the conjugation enzyme systems epoxide hydrolase, glucuronosyl transferase, sulphotransferase and glutathione S-transferase in precision-cut rat lung slices incubated in RPMI media for different time periods up to 72 h. Moreover, the effect of culturing of lung slices on total glutathione levels and glutathione reductase was also investigated. Monitoring of cytochrome P450 activity was achieved using established diagnostic probes, but when activity in the lung was low the maintenance of the various enzymes in culture was determined immunologically using Western blotting. The dealkylation of pentoxyresorufin declined markedly during the first 4h of incubation but in the case of ethoxyresorufin loss of activity was more gradual and less severe. Western blot analysis revealed that the rate of decrease in cytochrome P450 apoprotein levels was isoform-specific with CYP2E1 being the most stable and CYP3A the least stable. Generally, phase II activities, especially cytosolic sulphotransferase, were relatively more stable throughout the incubation period compared with cytochromes P450. Finally, glutathione reductase activity and total glutathione levels were maintained throughout the 72 h incubation. The present studies indicate that xenobiotic-metabolising enzymes in precision-cut rat lung slices decline in culture, but the rate of loss differs and depends on the nature of the enzyme. PMID- 16701935 TI - Lower blood pressure correlates with poorer performance on visuospatial attention tasks in younger individuals. AB - The relationship between low blood pressure and cognitive function among younger individuals is not fully understood. While a number of studies have examined hypertensive and hypotensive individuals, particularly in older populations, little attention has been devoted to healthy, young populations. We tested 105 healthy young individuals whose blood pressure levels naturally fell in the below normal-to-normal range. Our primary finding was a positive relation between blood pressure and cognition, as measured by two visuospatial attention tasks. This relation appears to be specific to visuospatial skills, as no relationship was observed between recognition memory and blood pressure. We discuss possible explanations for this positive relationship, such as structural neural mechanisms, and how they apply to the overall blood pressure-cognition relationship. PMID- 16701936 TI - Chlamydia trachomatis in subfertile couples undergoing an in vitro fertilization program: a prospective study. AB - OBJECTIVES: The objectives were to estimate the prevalence of Chlamydia trachomatis infection in subfertile couples and to study the relationship between markers of C. trachomatis infection and male infertility as well as pregnancy rates after in vitro fertilization (IVF). STUDY DESIGN: All consecutive couples consulting for infertility and IVF in Pellegrin Hospital were screened for C. trachomatis by direct (PCR test) and serological methods. RESULTS: Two hundred and seventy-seven couples were included in the study (mean age in years: 35 for men, 32 for women; mean duration of infertility: 4 years). The most frequent indication for IVF was tubal factor in 33%, endometriosis in 6%, dysovarian function in 12%, male infertility in 36% and others in 13%. C. trachomatis PCR was positive in 1.2% of men, 95% confidence interval (CI95%): (0.2%; 3.3%) and in 2.7% of women, CI95%: (1.1%; 5.5%). When combining all chlamydial markers, 17.3% of men, CI95%: (12.7%; 22.8%) and 20.4% of women, CI95%: (15.6%; 25.9%) had at least one positive marker. The presence of positive markers was not associated with altered semen characteristics. Couples with positive markers had a pregnancy rate of 23.1% (12 out of 52) compared with 20.2% (24 out of 119) among those with negative markers. CONCLUSION: In this population, the presence of past or current C. trachomatis infection was associated with neither semen characteristics nor outcome of IVF in subfertile couples. PMID- 16701937 TI - Variable path length and counter-flow continuous variation methods for the study of the formation of high-affinity complexes by absorbance spectroscopy. An application to the studies of substrate binding in cytochrome P450. AB - Studies of the equilibrium of protein-ligand interactions and determination of the stoichiometry of protein complexes constitute an important element of routine biochemical practice. In this paper we describe two innovative modifications of Job's method of continuous variation, which allow us to analyze tight interactions and determine stoichiometry in multi-site binding systems, including cases where the absorbance of the ligand overlaps with that of the enzyme-ligand complex. Our results on the interactions of cytochromes P450 3A4 and P450eryF with substrates illustrate the applicability of these approaches to the studies of substrate binding in enzymes that exhibit homotropic cooperativity. PMID- 16701938 TI - Guidelines and legislation for dye house effluents. AB - Increasing attention has been paid over the years to the environmental problems related to industrial activities. Environmental standards have been established and control organisms have been created with a view to applying restrictive legislation. In the dyeing industry, ennoblers discharge in the environment large volumes of strongly coloured effluents that are heavily loaded with pollutants and highly concentrated in salts. This chemical load is generated during the different steps of textile preparation. After reviewing the characteristics of these effluents, this paper gives a comparative review of the current legislations on the dyeing industry emissions. It examines more closely the discharge legislation, first in France and then in other countries around the world. A comparison of the six most representative parameters shows how different the applied constraints are in different countries. PMID- 16701939 TI - Prostaglandin receptors are mediators of vascular function in endometrial pathologies. AB - Prostaglandins are bioactive lipids produced from arachidonic acid by cyclooxygenase enzymes and specific terminal prostanoid synthase enzymes. Following biosynthesis, prostaglandins exert an autocrine/paracrine function by coupling to specific prostanoid G protein-coupled receptors to activate intracellular signaling and gene transcription. For many years prostaglandins have been recognised as key molecules in reproductive biology by regulating ovulation, endometrial physiology and proliferation of endometrial glands and menstruation. More recently a role for COX enzymes and prostaglandins has been ascertained in reproductive tract pathology, including dysmenorrhea, endometriosis, menorrhagia and cancer. Emerging evidence supports a role for COX enzymes, prostaglandins and prostaglandin receptor signaling pathways in a multitude of phenotypic changes in reproductive tissues including the promotion of angiogenesis and vascular function. Here we provide an overview of some of the findings from these studies with specific emphasis on the role of cyclooxygenase enzymes, prostaglandins and their receptors in benign and neoplastic pathologies of the human endometrium. PMID- 16701940 TI - LINAC radiosurgery for intracranial cavernous malformation: 10-year experience. AB - BACKGROUND: The annual hemorrhage rate of intracranial cavernous malformation (CM) is reported to range from 0.23% to 1.1%. Because of the low hemorrhage rate, operating on a deep symptomatic lesion with or without hemorrhage is considered controversial. For the prevention of hemorrhage, radiosurgery is an alternative method, targeting smaller lesions and delivering higher doses of radiation. Linear accelerator (LINAC) radiosurgery, aside from the gamma knife (GK), is not often discussed in the treatment of CM. PATIENTS AND METHODS: From 1995 to 2005, 30 patients presenting with hemorrhage or seizures, aged 14-79 years (mean 24.0 years) with single (27 patients) or multiple (three patients) CMs received LINAC radiosurgery at our institute. Six patients received LINAC radiosurgery following craniotomy for residual lesions revealed by the follow-up MRI. The temporal lobe was the most common site for CM in this series (n=8), followed by the brain stem (n=7). Thirty patients received 34 radiosurgery treatments with peripheral doses ranging from 800 cGy to 2200 cGy. The mean follow-up time was 59.9 months (range 1-122 months). RESULTS: One patient re-bled post-radiosurgery (0.67% in 149.75 observation-years). Two patients had asymptomatic post-LINAC edema (6.7%). Those three patients were symptom-free during the follow-up period. CONCLUSION: We suggest that LINAC radiosurgery is a relatively safe technique for treating deep or residual CM, although the actual effectiveness for CM obliteration is not known. PMID- 16701941 TI - An aggregate fuzzy hazardous index for composite wastes. AB - In this paper, a fuzzy waste index for evaluating the hazard posed by composite wastes generated from industrial processes is proposed. Within this methodology, a fuzzy index as a measure of hazardousness of a given composite waste is derived from the crisp inputs of its component's flammability, corrosivity, toxicity and reactivity attributes based on the National Fire Protection Association (NFPA) hazard rankings. The novelty of this work lies in establishing an integrated fuzzy hazardous waste index (FHWI) which provides a single-value representing the hazard ranking of a composite waste. This is contrary to current techniques which do not provide a final aggregated hazard index. The efficacy of the new proposed approach is illustrated through several worked examples. The results demonstrate that the fuzzy algorithm can be useful in aiding policy and decision-makers in conducting comprehensive initial evaluation of the status of waste hazardous status without the need for costly laboratory experiments. As such, the approach offers a robust and transparent decision-making methodology. PMID- 16701942 TI - A novel assessment tool for reusability of wastes. AB - This paper presents a new assessment method, which is able to determine the practicality of opportunities to reuse wastes. In this study, the term--reuse has much wider concepts than simply recycling to a process. In other words, reuse in this case can include recycling, utilisation as a raw material for other processes, and particularly transformation of the waste to low or preferably high value added products. Preliminary opportunities for the reuse of a waste can be generated by various methods such as brainstorming, desk research, consultation with industrial and academic contacts, and the like. In this work, each preliminary solution is then assessed according to three different categories- technical, economic, and environmental and regulatory, in order to determine the viability for reuse. The responses of nine collaborating companies to questionnaires pertaining to the three categories were used to set up feasible boundaries of each category in terms of the reuse of their wastes. Based on the replies, lower bounds for each category were determined, and then the preliminary solutions generated were ranked. The approach was applied to several industrial examples. The ranking of the high-dimensional information was aided by visual representation on a parallel coordinate graphic plot. Although the selection or rejection of an opportunity was highly dependent on the boundaries obtained, the approach proposed could provide a useful guideline to decision-makers for selection/rejection of the reuse opportunities available. PMID- 16701943 TI - Synthesis, characterization and thermolysis of 1,1-diamino-2,2-dinitroethylene (FOX-7) and its salts. AB - The present paper discusses the efforts made in HEMRL to establish the synthesis of FOX-7 at 100 g/batch level. In the present study, 1,1-diamino-2,2 dinitroethylene has been synthesised by treatment of acetamidinium chloride with diethylmalonate to obtain 2-methyl-pyrimidine-4,6-dione which on nitration followed by hydrolysis gave FOX-7. The synthesised FOX-7 has been characterized by spectroscopic and thermal techniques. The data obtained confirms the structure of FOX-7. The sensitivity of FOX-7 towards mechanical stimuli indicated its insensitive nature. The theoretically computed explosive and ballistic parameters are close to that of RDX. The synthesised FOX-7 has been used as a precursor for the synthesis of potassium and guanidinium salts and the thermal analysis of these salts indicate their exothermic nature. PMID- 16701944 TI - Microtubule-bundling activity of APC is stimulated by interaction with PSD-95. AB - Adenomatous polyposis coli (APC) tumor suppressor protein binds to microtubules, leading to microtubule bundling and stabilization. The protein also interacts with postsynaptic density (PSD)-95, a major scaffolding protein in neurons. Here, we analyzed the effects of PSD-95 on the microtubule-bundling activity of APC. The coexpression of APC and PSD-95 in COS-7 cells enhanced microtubule-bundle formation compared with the expression of APC alone. A mutant APC variant that does not associate with PSD-95 did not enhance microtubule bundling, despite coexpression with PSD-95. Immunoelectron microscopy showed that the APC-PSD-95 complex sometimes colocalized on microtubules in processes of cultured neurons. These results suggest that the microtubule-bundling activity of APC is regulated by its interaction with PSD-95, which might modulate microtubule architecture and dynamics in neurons. PMID- 16701945 TI - No association of attention-deficit/hyperactivity disorder with genes of the serotonergic pathway in Han Chinese subjects. AB - Attention-deficit/hyperactivity disorder (ADHD) is a complex psychiatric syndrome with cardinal symptoms of inattention, hyperactivity and impulsivity, and is a significant risk factor for poor health outcomes in both adolescence and adulthood. Etiology is clearly multifactoral, with probable contributions from both genetic and environmental factors. The genetic contribution is prominent, with estimated heritability at about 0.80. Although effects in dopamine metabolism have long been implicated in the etiology of ADHD, the role for serotonin has gained more attention in recent years. The current study examined five variants in three serotonin genes [those that code for serotonin receptors 2A (HTR2A), 5A (HTR5A) and 6 (HTR6)] in a relatively large sample of ADHD nuclear families. The transmission disequilibrium test (TDT) and the extended transmission disequilibrium test (ETDT) were performed to test for evidence of distorted transmission of alleles or haplotypes. No significant biased transmission was observed. These results do not support a substantial role of these serotonin gene in ADHD, however, additional work may be warranted before this association is definitively discounted. PMID- 16701946 TI - Cholesterol synthesis rate in human hippocampus declines with aging. AB - During the last three to four decades, interest in the interaction of circulating and brain cholesterol has increased. As the CNS matures and cholesterol pools in the brain become constant, the rate of de novo synthesis of cholesterol in the brain is expected to decline. We measured cholesterol, its precursors and its brain specific metabolite 24S-hydroxycholesterol in hippocampus from 7 female and 13 male corpses by highly sensitive and specific gas chromatography-mass spectrometry. Two age groups (young, n=10; elderly, n=10) were formed with a cut off at the median age of 38 years. The amount of cholesterol was comparable in young and elderly subjects. The concentrations of the cholesterol precursors lanosterol and lathosterol were significantly higher in young (P=0.036 and 0.005, respectively) than in elderly subjects. In accordance, there was a significantly negative correlation between age and lathosterol concentrations (r=-0.505; P=0.023). Absolute levels of 24S-hydroxycholesterol in the brain were slightly, but not significantly, lower in the hippocampal specimens from the elderly subjects. We conclude that during aging, cholesterol synthesis is decreased in the hippocampus, while absolute cholesterol content remains at a stable level. PMID- 16701947 TI - Moderate hypercapnia-induced anesthetic effects and endogenous opioids. AB - The purpose of this report is to explore the mechanisms of hypercapnia-induced antinociception. We carried out three experiments, the first to confirm whether moderate hypercapnia induces anesthetic effects, the second to determine whether naloxone reverses the anesthetic effects, and the third to evaluate whether beta endorphin is related to the anesthetic effects. In a pre-test, we determined the optimal CO(2) concentration in a chamber which would cause moderate hypercapnia in rats. Eighteen rats were divided into control, hypercapnia, and hypercapnia plus naloxone groups in experiment 1. The naloxone group rats were injected with naloxone (10 mg/kg) intraperitoneally before gas inhalation. After 60 min gas inhalation, 10% formalin was injected into the left rear paw of all rats, and nociceptive behaviors were observed for 1 h. In experiment 2, 11 rats were divided into control and hypercapnia groups. The brain was removed and fixed under pentobarbital anesthesia. Sections were immunostained for c-Fos and beta endorphin (ACTH) with the ABC method. All neurons double-labeled for c-Fos and beta-endorphin (ACTH) in the arcuate nucleus were counted by blinded investigators. Moderate hypercapnia (PaCO(2) 83+/-7 mmHg) reduced nociceptive behavior in the formalin test and naloxone pre-treatment attenuated this phenomenon. However, beta-endorphin-producing neurons were not activated by CO(2) inhalation. Endogenous opioids are related to moderate, hypercapnia-induced anesthetic effects, but, beta-endorphin-producing neurons in the hypothalamus were not activated by the CO(2) inhalation stress. PMID- 16701948 TI - Association study of the PIN1 gene with Alzheimer's disease. AB - Genetic linkage studies indicate evidence for one or more Alzheimer's disease (AD) genes on chromosome 19 independently of the apolipoprotein E gene, a well characterized AD-risk factor. Recently, the PIN1 gene on chromosome 19p13.2 has been proposed as a candidate gene for AD. Here, we have investigated the potential impact of two promoter polymorphisms (rs2233678 and rs2233679) within this gene on the risk of developing AD. No association of these polymorphisms or haplotypes with the disease was observed in a large French case-control population. Our data suggest that these genetic variants in PIN1 do not make a significant contribution to AD risk. PMID- 16701949 TI - Reproduction of self-rotation duration. AB - The vestibular system detects the velocity of the head even in complete darkness, and thus contributes to spatial orientation. However, during vestibular estimation of linear passive self-motion distance in darkness, healthy human subjects mainly rely on time, and they replicate also stimulus duration when required to reproduce previous self-rotation. We then made the hypothesis that the perception of vestibular-sensed motion duration is embedded within encoding of motion kinetics. The ability to estimate time during passive self-motion in darkness was examined with a self-rotation reproduction paradigm. Subjects were required to replicate through self-driven transport the plateau velocity (30, 60 and 90 degrees /s) and duration (2, 3 and 4s) of the previously imposed whole body rotation (trapezoid velocity profile) in complete darkness; the rotating chair position was recorded (500 Hz) during the whole trials. The results showed that the peak velocity, but not duration, of the plateau phase of the imposed rotation was accurately reproduced. Suspecting that the velocity instruction had impaired the duration reproduction, we added a control experiment requiring subjects to reproduce two successive identical rotations separated by a momentary motion interruption (MMI). The MMI was of identical duration to the previous plateau phase. MMI duration was fidelitously reproduced whereas that of the plateau phase was hypometric (i.e. lesser reproduced duration than plateau) suggesting that subjective time is shorter during vestibular stimulation. Furthermore, the accurate reproduction of the whole motion duration, that was not required, indicates an automatic process and confirms that vestibular duration perception is embedded within motion kinetics. PMID- 16701950 TI - Kainate exposure suppresses activation of GluR2 subunit promoter in primary cultured cerebral cortical neurons through induction of RE1-silencing transcription factor. AB - The AMPA receptor subunit GluR2 is downregulated in neurons following a wide range of neurological insults. Here we report that suppression of GluR2 gene promoter activity is associated with kainate (KA)-induced downregulation of GluR2 subunit levels in primary cultured cortical neurons. RT-PCR and Northern blotting showed a significant decrease in GluR2 mRNA in cultured neurons after KA exposure. Transfection of cultured neurons with an expression vector pGL3-GluR2( 298/+283), where the reporter gene firefly luciferase was driven by the GluR2 promoter, revealed that KA exposure suppressed the transcriptional activation of the GluR2 promoter. Furthermore, the expression of the RE1-silencing transcription factor (REST) was increased in KA-exposed cortical neurons; enhanced binding of REST to RE1-like silencer element in the proximal promoter of the GluR2 subunit gene was evidenced by electrophoresis mobility shift assay. Chromatin immunoprecipitation showed that suppressed activity of the GluR2 promoter in cultured neurons after KA exposure was related to deacetylation of histone H4. These results indicate that REST as a crucial factor binds to RE1 like silencer element in the GluR2 promoter, suppressing transcription of the GluR2 subunit gene during KA exposure. Our data suggest that transcriptional suppression of the GluR2 subunit gene may contribute at least in part to downregulation of GluR2 subunit protein in neurons during KA exposure. Because our experiments showed a reduction of glutamate release in KA-exposed cortical neurons, REST may play a latent role in delayed neuronal death or in seizure induced tolerance. PMID- 16701952 TI - How prior experience shapes placebo analgesia. AB - Some studies indicate that placebo analgesia is stronger when pre-conditioning with effective analgesic treatments is performed, thereby suggesting that the placebo response is a learning phenomenon. Here we further tested this hypothesis in order to better understand when and how previous experience affects the placebo analgesic response. To do this, we used a conditioning procedure whereby the intensity of painful stimulation was reduced surreptitiously, so as to make the subjects believe that an analgesic treatment was effective. This procedure induced strong placebo responses after minutes, and these responses, albeit reduced, lasted up to 4-7 days. In addition, in a second group of subjects we repeated the same conditioning procedure 4-7 days after a totally ineffective analgesic treatment, and found that the placebo responses were remarkably reduced compared to the first group. Thus we obtained small, medium and large placebo responses, depending on several factors, such as the previous positive or negative experience of an analgesic treatment and the time lag between the treatment and the placebo responses. We also ran extinction trials, and found that these effects did not undergo extinction in a time span of several minutes. These findings indicate that placebo analgesia is finely tuned by prior experience and these effects may last, albeit reduced, several days. These results emphasize that the placebo effect is a learning phenomenon in which many factors come into play, and may explain the large variability of the placebo responses that is found in many studies. PMID- 16701951 TI - Enhanced neurogenesis in the ischemic striatum following EGF-induced expansion of transit-amplifying cells in the subventricular zone. AB - In the subventricular zone (SVZ) of the adult mammalian brain, neural stem cells continually produce transit-amplifying precursors, which generate neuroblasts migrating into the olfactory bulb. Previous studies have suggested that SVZ cells also have the capacity to generate some striatal neurons after cerebral ischemia. The infusion of epidermal growth factor (EGF) has been demonstrated to increase the number of these regenerated neurons. However, which cell types in the SVZ are stimulated to proliferate or differentiate after EGF infusion remains unknown. In this paper, we demonstrated that cerebral ischemia results in an increase in the number of EGF receptor (EGFR)-positive transit-amplifying cells in the SVZ. EGF infusion into the ischemic brain caused the number of transit-amplifying cells to increase and the number of neuroblasts to decrease. On the other hand, after an interval of 6 days after the discontinuation of EGF infusion, a significant increase in the number of neuroblasts was found, both in the striatum and the SVZ. These results suggest that the replacement of neurons in injured striatum can be enhanced by an EGF-induced expansion of transit-amplifying cells in the SVZ. PMID- 16701953 TI - Decreased sensitivity to self-inflicted pain. AB - There is anecdotal and incidental research evidence suggesting that self inflicted injury is experienced as less painful than when the same injury is applied by another person. We tested the hypothesis that the sensitivity and the ability to tolerate pain differs depending on the person applying the painful stimulus. Self-selected healthy undergraduate students were obtained from the University of Stirling participant panel. None were suffering chronic pain or taking any form of analgesic drug. The participants applied a pressure algometer to themselves and to other participants. Depending on the type of trial, each was terminated when the participant experienced the algometer as either "painful" (for threshold reading) or "unbearable" (for tolerance reading). Both measures of pain, threshold and tolerance, were significantly higher when the algometer had been self-applied compared with when it was applied by another person. The mean difference for pain thresholds was 0.27MPa (95% confidence interval 0.10-0.44, P=0.002), and the mean difference for pain tolerance readings was 0.25MPa (95% confidence interval 0.03-0.48, P=0.028). An unexpected finding was that the mean tolerance score was less when females applied the algometer (P<0.01). When a painful stimulus was self-inflicted this resulted in significantly less pain and a greater ability to tolerate the pain compared with when the same stimulus was applied by another person. If the findings generalized to a clinical context, certain painful or discomforting procedures, such as mammography, removal of wound dressings and lancet withdrawal of blood, should be adapted for self application by patients. PMID- 16701955 TI - Transmission properties of hydrogen and helium ion implantation induced waveguide grating structures and potential application. AB - Optical waveguides in fused silica have been manufactured by H(+) implantation, which shows significantly improved transmission properties after implantation. The relationship between the transmission property and the thickness of the cladding layer after surface etching has also been investigated. Grating structures in waveguides have been made by additional He(+) implantation with a periodic metal mask covering the surface. The transmission of such grating structures is leveled throughout the measured wavelength range. This observation suggests a new method to make variable optical attenuators (VOAs) to equalize optical powers of different channels. PMID- 16701954 TI - A twin study of posttraumatic stress disorder symptoms and chronic widespread pain. AB - Previous studies of the association between posttraumatic stress disorder (PTSD) and chronic widespread pain (CWP) or fibromyalgia have not examined the role of familial or genetic factors. The goals of this study were to determine if symptoms of PTSD are related to CWP in a genetically informative community-based sample of twin pairs, and if so, to ascertain if the association is due to familial or genetic factors. Data were obtained from the University of Washington Twin Registry, which contains 1042 monozygotic and 828 dizygotic twin pairs. To assess the symptoms of PTSD, we used questions from the Impact of Events Scale (IES). IES scores were partitioned into terciles. CWP was defined as pain located in 3 body regions lasting at least 1 week during the past 3 months. Random effects regression models, adjusted for demographic features and depression, examined the relationship between IES and CWP. IES scores were strongly associated with CWP (P<0.0001). Compared to those in the lowest IES tercile, twins in the highest tercile were 3.5 times more likely to report CWP. Although IES scores were associated with CWP more strongly among dizygotic than among monozygotic twins, this difference was not significant. Our findings suggest that PTSD symptoms, as measured by IES, are strongly linked to CWP, but this association is not explained by a common familial or genetic vulnerability to both conditions. Future research is needed to understand the temporal association of PTSD and CWP, as well as the physiological underpinnings of this relationship. PMID- 16701956 TI - A survey of mutations in the Cochliomyia hominivorax (Diptera: Calliphoridae) esterase E3 gene associated with organophosphate resistance and the molecular identification of mutant alleles. AB - Cochliomyia hominivorax (Calliphoridae) is one of the most important myiasis causing flies and is responsible for severe economic losses to the livestock industry throughout the Neotropical region. In Brazil, C. hominivorax has been controlled mainly with organophosphate (OP) insecticides, although the inappropriate use of these chemicals can result in the selection of resistant flies. Changes in carboxylesterase activity have been associated with OP insecticides in some arthopodan species. In this work, we isolated and characterized part of the E3 gene in C. hominivorax (ChalphaE7), which contained the same substitutions responsible for the acquisition of OP hydrolase activity in Lucilia cuprina (Calliphoridae). Digestion of the polymerase chain reaction products with a restriction enzyme that specifically recognized the mutation site unambiguously differentiated wild and mutated esterase alleles. The PCR-RFLP assay therefore provided a fast, reliable DNA-based method for identifying C. hominivorax individuals with a mutation in the esterase gene. Further bioassays to determine the association of this mutation with OP resistance in C. hominivorax should allow the development of more effective strategies for managing this species. PMID- 16701958 TI - Despite their inevitable conflicts--science, religion and New Age spirituality are essentially compatible and complementary activities. AB - Until recently it seemed that the continued expansion of scientific ways of thinking was destined to render religion extinct and spirituality unfeasible. But the example of the United States disproves this, since America is the most successful scientific nation of this era, church-going remains strong and New Age spiritualities are thriving. Therefore, despite the obvious conflicts; science, religion and spirituality are essentially compatible. Future science will continue to win territory from religion since its validation procedures are more objective and reliable. However, churches can survive and grow by dropping those aspects of doctrine which clash with science, and expanding their social functions. The fast-growing US 'mega-church' movement shows the way - since these organizations are minimally dogmatic but instead provide a family-orientated and morally-cohesive social milieu. Like organized religion, New Age spirituality comes into conflict with science when it makes incredible or bizarre factual claims. However, in practice modern spirituality is based on subjective evaluations which do not clash with the procedures of science. Indeed, the reliance upon individual, emotion-based evaluations (e.g., 'my truth', 'whatever works for you') renders New Age spirituality 'science-proof', and has enabled it to expand massively in an age of science. Science, religion and spirituality perform different functions in the modern world, and their relationship is therefore one of mutual-dependence. Borderline disputes will inevitably occur, but as part of a broader context of complementarity. Science, 'social' churches and New Age spirituality all have a bright future. PMID- 16701957 TI - The dex/CRH test--is it better than the DST? AB - The dexamethasone suppression test (DST), frequently abnormal in mood disorder patients, is considered to measure glucocorticoid receptor-mediated negative feedback. We examined the hypothesis that the, apparently more sensitive, dexamethasone/corticotrophin-releasing-hormone (dex/CRH) test unveils subtle hypothalamic-pituitary-adrenal axis disturbance not detected by the DST in 82 patients with mood disorders and 28 controls. There was a close correlation between the cortisol responses on the two tests (r(s)=0.73, p<0.0005). However, ROC analysis revealed that the dex/CRH test had better diagnostic performance than the DST (p=0.031). The sensitivity of delta cortisol (from the dex/CRH) was 61.9% and the specificity 71.4%. The sensitivity of 1500 h cortisol (the DST) was 66.6% and the specificity was 47.6%. This suggests that the two tests measure common pathology but that the dex/CRH test has better diagnostic utility. PMID- 16701960 TI - Peripheral arterial disease: a manifestation of evolutionary dislocation and feed forward dysfunction. AB - Peripheral arterial disease in the legs represents a subset of atherosclerosis that manifests a particularly sinister profile. A predominance of sympathetic activity in the periphery favors the development of neurogenic atherosclerosis. Atherosclerosis may then produce flow derangements and decreased physical activity that serves to escalate sympathetic bias in a vicious cycle. Restoration of normal flow in peripheral arterial disease may not only produce local benefit due to improved perfusion, but also represent a gateway to correcting many systemic conditions that may at first glance appear unrelated but share a common etiology of autonomic dysfunction, such as gout, acute coronary syndromes, stroke, sleep apnea, arrhythmias, depression, erectile dysfunction, inflammation, hypercoagulability, sleep disorders, bowel dysfunction, renal failure, and aging. PMID- 16701959 TI - Solar cycles and their relationship to human disease and adaptability. AB - In this paper, we show that 11-year solar cycle peaks predispose humans to disease, but also endow creativity and adaptability. We give several examples of diseases that are modulated by light and present evidence for an effect of intensity and variation in sunlight, primarily ultraviolet radiation (UVR), on the human genome. The birth dates of nearly 237,000 unique clients in the Maine Medicaid database collected from 1995 to 2004, inclusive, were related to solar cycle irradiance for the past seventy-one years, encompassing seven solar cycles. The sample was divided into four general categories of disease: mental/behavioral illnesses; metabolic diseases; autoimmune diseases; neoplasms. The birth months for those clients born in any given year were arranged in the form of a winter/summer ratio in order to more clearly appreciate the seasonality inherent in each disease category. Solar cycles were separated into chaotic (approximately three times as irradiant) or non-chaotic according to the Gutenberg-Richter power law and the uncertainty inherent in predicting solar storms. The results show that radiation peaks in solar cycles and particularly in chaotic solar cycles (CSCs) are associated with a higher incidence of mental disorders, suggesting the sensitivity of ectodermal embryonic tissues to UVR. Autoimmune diseases have intermediate sensitivity, while the neoplasms in the study, primarily of endoderm, appear suppressed by peak UVR intensity. The ratio of the number of clients born in CSC cycles to non-CSC cycles was highest for the more genetic mental diseases, like schizophrenia and bipolar disorder, but as that ratio decreased, the clients with diseases like multiple sclerosis and rheumatoid arthritis showed more environmental features manifested as a greater winter/summer birth month ratio that was significantly different than that of the average client in the whole data set. The paper presents evidence that latitude, e.g., variation in light, is an added stress to the immune system (especially at 53-54 degrees N. latitude) that is involved in nearly all human disease. We hypothesize that introns, the presumptive engenderers of gene control, modulate the effects of UVR, particularly for the neoplasms studied. We conclude that intermittent and largely unpredictable peak solar cycle radiation has been the fundamental engine of evolution, forcing organisms to adapt to mutagenic UVR and producing enough damage to instigate genetic variation. Probably a chance genetic mutation over 80,000 years ago produced a human brain capable of abstract thought and consciousness. The slight genetic instability that favored an adaptable, creative brain also produced other somatic variations that present phenotypically as disease, but largely expressed after natural selection (reproduction) and associated with the inexorable entropy of aging. PMID- 16701961 TI - Antioxidative and acute antiinflammatory effects of Torreya grandis. AB - The present study was undertaken to investigate the antioxidative and antiinflammatory activities of the ethanolic extract of seeds of Torreya grandis (EST). Exposure of human dermal fibroblasts to the extract at 50 and 250 microg/ml showed significant protective effect against hydrogen peroxide (300 microM). EST not only protected cell survival from H(2)O(2)-induced toxicity, but also inhibited the H(2)O(2)-induced LDH release significantly. It was also found that EST at 100 and 1000 microg/ml showed scavenging activities of radicals and reactive oxygen species with 29.8% and 100.0% of inhibition against DPPH radical and 41.2% and 98.4% against superoxide radicals in the xanthine/xanthine oxidase system, respectively. Topically applied EST dose-dependently inhibited arachidonic acid (AA)- and 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced ear edema in mice. PMID- 16701962 TI - Antioxidant and 15-lipoxygenase inhibitory activity of rotenoids, isoflavones and phenolic glycosides from Sarcolobus globosus. AB - From Sarcolobus globosus, two rotenoids (villosinol and 6-oxo-6a,12a dehydrodeguelin), one isoflavone (genistin) and four phenolic glycosides (vanillic acid 4-O-beta-d-glucoside, glucosyringic acid, tachioside and isotachioside) were identified for the first time from this species. Extracts and compounds from S. globosus were evaluated for their DPPH radical scavenging and 15-lipoxygenase (15-LO) inhibitory activities. All tested rotenoids were found to inhibit 15-LO, while they lacked DPPH radical scavenging effect. PMID- 16701963 TI - Selective processing of cannabis cues in regular cannabis users. AB - Recent studies indicate that the regular use of certain drugs, such as tobacco and alcohol, is associated with biases in the processing of drug-related cues, as those cues grab attention, elicit approach and are perceived as pleasant. This study investigated whether regular cannabis users exhibit comparable cognitive biases for cannabis-related pictorial cues. Twenty-three regular cannabis users and 23 non-user controls completed a series of tasks including a visual probe task with concurrent eye movement monitoring (to measure attentional bias), a stimulus-response compatibility task (to measure implicit approach bias) and a valence rating task (to measure the perceived pleasantness of cannabis cues). Results indicated that, relative to non-users, regular cannabis users had biases to maintain their gaze on cannabis cues, to make faster approach responses towards cannabis cues, and to rate cannabis cues as pleasant. Results are generally consistent with previous findings from tobacco smokers and heavy drinkers, and the implications for incentive-motivational theories of addiction are discussed. PMID- 16701964 TI - Vaccine side effects: fact and fiction. AB - The debate over adverse reactions associated with companion animal vaccination has considerably exercised the veterinary profession internationally over the past decade. A range of suspected adverse reactions to vaccines is reported including the onset of inflammatory, allergic, autoimmune or neoplastic diseases. Lack of efficacy, interference with diagnostic testing and other occasional suspected product-related issues are also reported. Available data suggest that the overall prevalence of true adverse reactions is exceedingly low and that vaccination does not significantly contribute to ill-health in companion animals. There is increasing public interest in vaccination issues with transfer of focus from publicity over human vaccine side effects to those perceived to occur in animals. We must not lose sight of the fact that vaccination is a safe procedure that has impacted significantly on infectious disease control. Reduced population uptake of vaccination leads to re-emergence of disease in both humans and animals. Nevertheless, there have recently been a series of practical recommendations produced to ensure reduced 'vaccine load' on our companion animals and vaccine manufacturers are moving towards developing non-adjuvanted products with an extended duration of immunity. These measures will further reduce the very small current risk of any adverse consequences to vaccination in our pet population. PMID- 16701965 TI - New understanding of immunological mechanisms. AB - The understanding and importance of antigen-specific immune responses after vaccination has completely changed in recent years. In the past, the focus for monitoring a vaccine-specific immune reaction was principally on the humoral branch of the immune system. The efficacy of vaccines, as assessed by the induction of protective immunity was mainly correlated with antibodies and antibody-titers. However, this correlation often failed and other parts of the immune system had also to be considered: namely, the innate immune system and the cellular branch of the antigen-specific immune system. With regard to vaccines, the innate immune system plays its main role in the effective activation of the antigen-specific immune response, in antigen-uptake and antigen-presentation. The dendritic cells (DCs) are the most important antigen presenting cells which present processed protein antigens (peptides) through MHC-molecules: MHC-class I, for the presentation of endogenous synthesised antigen; MHC-class II for exogenous antigen. Activation of DC leads to an enhanced production of cytokines and chemokines, to an up-regulation of co-stimulatory and activation molecules and also molecules for cell-cell interactions, e.g. interactions with cells of the antigen-specific immune system. T lymphocytes are the effector cells of the cellular branch of the antigen-specific immune system. They act either as MHC class I-restricted cytolytic T lymphocytes (CTL) or as MHC-class II-restricted T helper cells providing support for B lymphocytes (T(H)2) and the cellular part of the antigen-specific immune system (T(H)1). In order to achieve effective vaccination, the activation of all T-cell subpopulations is of advantage, but more important is the generation of antigen-specific memory T and B lymphocytes. In addition to these 'generic' immunological factors which are essential for the design of more efficacious vaccines, our detailed knowledge about feline and canine immune reactions after vaccination, which is still poor, has to be improved. PMID- 16701966 TI - Canine vaccination--providing broader benefits for disease control. AB - This paper reviews the broader benefits of canine vaccination to human and animal health and welfare with an emphasis on the impacts of mass dog vaccination against rabies in countries of the less-developed world. Domestic dogs are the source of infection for the vast majority (>95%) of cases of human rabies worldwide, and dogs remain the principal reservoir throughout Africa and Asia. Canine vaccination against rabies has been shown to dramatically reduce the number of cases in dogs, the incidence of human animal-bite injuries (and hence the demand for costly post-exposure prophylaxis) and the likely number of human cases, primarily in children. Further benefits include the mitigation of the psychological consequences of rabies in a community, improved attitudes towards animals and animal welfare and reduced livestock losses from canine rabies. Mass vaccination has recently been used in the conservation management of wild carnivore populations threatened by transmission of rabies and canine distemper virus from domestic dog populations. Vaccination of wildlife hosts directly may also provide an option for mitigating infectious disease threats. The development of integrated control measures involving public health, veterinary, wildlife conservation and animal welfare agencies is needed to ensure that control of canine diseases becomes a reality in Africa and Asia. The tools and delivery systems are all available--all that is needed is the political will to free the world from the ongoing tragedy of these diseases. PMID- 16701967 TI - Molecular detection of porcine enteric caliciviruses in Venezuelan farms. AB - Caliciviruses are a well-established cause of respiratory, vesicular and hemorrhagic diseases in animals. In addition, these viruses are an important cause of enteric diseases in humans. Recently, molecular analysis of several porcine enteric caliciviruses indicated that they are closely related to human enteric caliciviruses. The objective of this work was to determine the frequency, age distribution, and association with diarrhea of enteric calicivirus infections in piglets and to partially characterize the detected isolates. A total of 203 stool samples from animals 0 to 9 weeks of age, collected between 1993 and 2003 in seven porcine farms located in the central region of Venezuela were tested for enteric caliciviruses by reverse transcription-polymerase chain reaction (RT-PCR) amplification using primers designed to detect both norovirus and sapovirus. Selected amplicons were sequenced to establish phylogenetic relationships with reference strains. Calicivirus were detected in 18% (36/204) of the samples. Viruses were detected more frequently in animals between 3 and 4 weeks of age, and were detected in samples from animals with diarrhea and without diarrhea with equal frequencies (14 versus 19%, p>0.5). Phylogenetic studies based on partial RNA polymerase gene sequences indicated that the Venezuelan isolates were most closely related (75-95% identity) to the sapovirus Cowden reference strain. These results provide evidence that caliciviruses of the genus sapovirus circulate frequently in piglets but further studies are needed to clarify their importance as cause of diarrhea. PMID- 16701969 TI - The combined use of human neural and liver cell lines and mouse hepatocytes improves the predictability of the neurotoxicity of selected drugs. AB - The cytotoxicity of amitriptyline (0-100microM), selegiline (0-4.5microM), carbamazepine (0-420microM) and paracetamol (0-10mM) was studied in metabolically competent mouse hepatocytes, metabolically incompetent human hepatoblastoma (HepG2) cells, and in neuroblastoma (SH-SY5Y) and astrocytoma (U-373 MG) cells, by using luminescence-based ATP measurement as an endpoint of cell toxicity. The aim was to evaluate the potential of the selected cell cultures to recognize metabolism-induced toxicity of the test compounds, and to predict further hepatic and neural toxicity. In SH-SY5Y cells amitriptyline was severely toxic, while selegiline and paracetamol failed to show any toxic effect, and carbamazepine was only slightly toxic at the highest concentration. In U-373 MG cells the onset of amitriptyline toxicity started earlier than in SH-SY5Y cells. However, the highest amitriptyline concentration resulted in approximately 100% decrease in the viability of the SH-SY5Y cells, whereas the decrease in the viability of the U-373 MG cells was only approximately 30%. Selegiline, carbamazepine and paracetamol were toxic in mouse hepatocytes (but not in HepG2 cells), which suggests that these drugs may show metabolism-dependent (neuro)toxicity. In conclusion, compared to the use of neurons alone, better estimations of neurotoxicity can be made by the combined use of metabolically competent hepatocytes and glial cells (e.g. U-373 MG) together with neuronal cells (e.g. SH SY5Y). PMID- 16701971 TI - Novel design of osmotic chitosan capsules characterized by asymmetric membrane structure for in situ formation of delivery orifice. AB - In this study, chitosan capsules with asymmetric membrane to induce osmotic effects and in situ formation of the delivery orifice were optimally prepared and characterized. Chitosan capsules were formed on stainless steel mold pins by dipping the pins into a chitosan solution followed by forming asymmetric structure by dipping into a quenching solution containing tripolyphosphate (TPP) to cause an ionic cross-linking reaction between the outer layer of chitosan and TPP. Factors influencing the properties of the capsule membrane, such as the molecular weight of chitosan, the dipping solution and dipping time, and the quenching solution and time, were optimized to successfully produce osmotic chitosan capsules with asymmetric membrane using chitosans that possessed different viscosities. In situ formation of a delivery orifice on the asymmetric membrane of the chitosan capsule was proven by the observation of a jet stream of chlorophyll being released from the capsule. Drugs with different solubility were selected, and a linear correlation between drug solubility and the initial drug release rate calculated from the slope of the drug release profile was used to verify that the delivery orifices that were in situ formed on the asymmetric membrane of the chitosan capsules induced by osmotic effect was responsible for the drug release. Water permeability across the optimally produced asymmetric membrane of the capsule from chitosan of 500 cps (300-700 cps) quenched with TPP for 30 min (C500/TPP30) was determined to be 1.40 x 10(-6)cm(2)h(-1)atm(-1) at 37.0+/-0.5 degrees C. The encapsulation of poorly water-soluble drugs, felodipine (FE) and nifedipine (NF), in such an asymmetric chitosan capsule was capable of creating a sufficient osmotic effect to activate the release of the drug with the addition of SLS and HPMC. The multiple regression equations of maximal release percent at 24h for FE and NF confirmed that both sodium lauryl sulfate (SLS) and hydroxypropyl methylcellulose (HPMC) positively influenced this response factor, and the effect of SLS was greater than that of HPMC. PMID- 16701972 TI - Induction of systemic and mucosal immune responses by intranasal administration of alginate microspheres encapsulated with tetanus toxoid and CpG-ODN. AB - In the induction of systemic and mucosal immunity, particulate antigens are more effective than soluble antigens; possibly because they are more efficiently endocytosed by mucosal-associated lymphoid tissue (MALT) M cells. In this study, we determined the systemic and mucosal immune responses in rabbits following intranasal immunization with encapsulated tetanus toxoid (TT) and CpG-ODN in alginate microspheres. The microspheres were less than 4 microm in diameter. Encapsulation efficiency of TT and CpG-ODN was determined as 47.7+/-6.6 and 34.2+/-7.4, respectively. Release of TT and CpG-ODN in a simulated model with nasal cavity was 14.2+/-3.06 and 36.7+/-2.4% after 4 h. Encapsulated TT preserved its intact structure, but its immunoreactivity was decreased to about 91+/-5%. The highest serum IgG and antitoxin, and nasal lavage IgA titers were observed in groups immunized with microsphere formulations. CpG-ODN as an adjuvant could increase the serum IgG and antitoxin titers when co-administered with TT solution, but its co-encapsulation with TT in alginate microspheres failed to potentiate the systemic immune response while induced high IgA titers in nasal lavages. No hemolysis was occurred on incubation of alginate microspheres and human RBCs. Also after nasal administration of plain microspheres to human volunteers, no local irritation was observed. Intranasal administration of microspheres encapsulated with vaccines showed to be an effective way for inducing a variety of immune responses and that a strong systemic IgG and mucosal IgA responses can be induced in rabbits with intranasal administration of alginate microspheres encapsulated with TT. PMID- 16701973 TI - The delivery of ketoprofen from a system containing ion-exchange fibers. AB - A postulated model for transdermal delivery using ion-exchange fibers as controlling device was designed, and the main objective of this study was to assess the rationality of the model. The release rates of ketoprofen from the carbopol-based gel vehicles containing ion-exchange fibers to which the ketoprofen had been bound have been determined across 0.22 microm microporous membrane. The fluctuation of the release rate of ketoprofen from the vehicles was much lower compared with that of simple gels, though the cumulative amount of ketoprofen delivery was less. Additional ions could increase the rate and extent of ketoprofen delivery. The iontophoretically assisted transport of ketoprofen across rat skin was also studied and found to be favorable to ketoprofen permeation. According to the tested model, the ion-layer could enhance the ketoprofen delivery and satisfactory results were achieved. PMID- 16701974 TI - Implication of inclusion complexation of glimepiride in cyclodextrin-polymer systems on its dissolution, stability and therapeutic efficacy. AB - The effect of complexation of glimepiride, a poorly water-soluble antidiabetic drug, with beta-cyclodextrin and its derivatives (HP-beta-CyD and SBE-beta-CyD) in presence of different concentrations of water-soluble polymers (HPMC, PVP, PEG 4000 and PEG 6000) on the dissolution rate of the drug has been investigated. The results revealed that the dissolution rate of the drug from these ternary systems is highly dependent on polymer type and concentration. The dissolution rate of the drug from ternary systems containing PEG 4000 or PEG 6000 seems to be generally higher than from systems containing HPMC or PVP. An optimum increase in the dissolution rate of the drug was observed at a polymer concentration of 5% for PEG 4000 or PEG 6000 and at 20% concentration of HPMC or PVP. The dissolution rate of the drug from the ternary system glimepiride-HP-beta-CyD-5% PEG 4000 was high compared to the other systems. Tablets containing the drug or its equivalent amount of this ternary system were prepared and subjected to accelerated stability testing at 40 degrees C/75% R.H. to investigate the effect of storage on the chemical stability as well as therapeutic efficacy of the tablets. The results revealed stability of the tablets and consistent therapeutic efficacy on storage. PMID- 16701975 TI - Cholinergic neurons degenerate when exposed to conditioned medium of primary rat brain capillary endothelial cells: counteraction by NGF, MK-801 and inflammation. AB - Alzheimer's disease is characterized by extracellular beta-amyloid plaques, intraneuronal Tau-inclusions and cell death of cholinergic neurons. Recent evidence indicates that the vascular system may play an important role in the development of this progressive neurodegenerative disease. The aim of this study was to observe, if brain capillary endothelial cells (BCEC) may produce and secrete factors which induce cell death of cholinergic neurons, and if this effect is counteracted by (1) NGF, MK-801 or vitamin C, (2) modulated by experimentally-induced inflammation with interleukin-1beta and lipopolysaccharide (IL-1beta and LPS) or (3) by blocking of different intracellular signalling pathways. Cholinergic neurons were cultivated in organotypic brain slices of the nucleus basalis of Meynert and treated with conditioned medium derived from BCEC, supplemented with different protective factors. BCEC were stimulated with IL 1beta and LPS or different intracellular pathway inhibitors before collection of conditioned medium. Cholinergic neurons were detected by immunohistochemistry for choline-acetyltransferase. Possible effects on BCEC viability and proliferation were determined by nuclear staining, BrdU incorporation and release of nitrite and lactate-dehydrogenase. BCEC released factors that can kill cholinergic neurons. This neurotoxic effect was blocked by NGF and MK-801 (a NMDA antagonist), but not by vitamin C. Pretreatment of BCEC with intracellular pathway inhibitors did not change the neurotoxicity, but pretreatment with IL 1beta and LPS abolished the neurotoxic effect. In summary, BCEC produce and secrete molecules which induce excitotoxic cell death of cholinergic neurons. It is concluded that excitotoxic factors secreted by vascular cells may contribute to the development of cholinergic neurodegeneration as it occurs in Alzheimer's disease. PMID- 16701976 TI - Neuroprotective effect of etomidate on functional recovery in experimental spinal cord injury. AB - OBJECTIVE: Primary impact to the spinal cord causes rapid oxidative stress after injury. To protect neural tissue, it is important to prevent secondary pathophysiological mechanisms. Etomidate, a strong antiexcitotoxic agent, stimulates the gamma aminobutyric acid (GABA) receptors. The purpose of this study was to investigate neurobehavioral and histological recovery and to evaluate the biochemical responses to treatment of experimental spinal cord injury (SCI) in rats with etomidate or methylprednisolone (MP) or both etomidate and MP. MATERIAL AND METHODS: Seventy-two rats were randomly allocated into six groups: a control group (laminectomy alone), a trauma group (laminectomy+trauma), a methylprednisolone group (30 mg/kg MP), an etomidate group (2 mg/kg), a methylprednisolone and etomidate combined treatment group (30 mg/kg MP and 2 mg/kg etomidate) and a vehicle group. Six rats from each group were killed at the 24th hour after the injury. Malondialdehyde, glutathione, nitric oxide and xanthine oxidase levels were measured. Neurological functions of the remaining rats were recorded weekly. Six weeks after injury, all of those rats were killed for histopathological assessment. RESULTS: Etomidate treatment revealed similar neurobehavioral and histopathological recovery to MP treatment 6 weeks after injury. Combined treatment did not provide additional neuroprotection. CONCLUSION: Etomidate treatment immediately after spinal cord injury has similar neuroprotection to MP. In spite of different neuroprotection mechanisms, combined treatment with MP and etomidate does not provide extra protection. PMID- 16701978 TI - [Initial training in obstetric anaesthesia]. AB - Obstetric anaesthesia is a "young" discipline, with constant novelties from a clinical, scientific and academic standpoint. While there is still no official sub-specialty in obstetric anesthesia, this field has become more diversified because of the growing maternal request for labor analgesia, a constant and perhaps even increasing rate of caesarean deliveries of 20-30% depending on institutions and countries, and also due to the raise in "high risk" pregnancies in women carrying various medical conditions such as complex congenital cardiopathies. In their anesthesia training, most residents rotate in the delivery room for three months on average, which should allow them to acquire good practical skills when performing regional analgesia and anesthesia for labor and delivery. It has been shown that performing 75 epidurals in the obstetric context is sufficient to provide good technical expertise, when appropriate supervision is provided by a present and devoted staff. General anaesthesia remains the critical point, because this technique is less and less performed in pregnant women. It should be recommended to develop training programs to improve the technical skills for intubation in pregnant women, which is why anaesthesia simulators may have an important role in the future. In terms of the theoretical knowledge, good academic training programs are required. The physiology of pregnancy and the physiopathology of pregnancy-related disorders justify a thorough and rigorous teaching in order to reduce both maternal and neonatal morbidity and mortality. PMID- 16701977 TI - [Pelvic pain by acute symphysis pubis separation after vaginal delivery]. AB - Pelvic pain during pregnancy and postpartum period is common. Pubic separation is a physiologic phenomenon caused by pregnancy and delivery. It's an aetiology of pregnancy and postpartum pain requiring a specialized management in case of severe pain. We report the case of a 34-year-old multiparous woman suffering from severe pubic symphysis pain after twin delivery by vaginal approach, with extraction support, under epidural analgesia. Pubic symphysis separation was diagnosed according both to the clinical symptoms and to the radiology. A local anesthetic infiltration was performed twice to substantially relieve the patient. PMID- 16701979 TI - [Brain perfusion and metabolism imaging techniques]. AB - Due to recent efforts in improving spatial and temporal resolution in imaging techniques, it is now possible to get relevant information about brain perfusion and metabolism in humans. This information can significantly impact on brain pathophysiology, diagnosis assessment and therapy options, particularly in patients having brain ischemia. Among these imaging and metabolism techniques are dynamic perfusion computed tomography, perfusion MRI, positron emission tomography and NMR spectroscopic imaging. The goal of this article is an overview of these four techniques, with their own technical description, advantages and drawbacks. Details are provided about brain parameters given by each technique and their clinical relevance, the accessibility of the technique in the emergency setting and the optimal window to use it during the patient's evolution. PMID- 16701980 TI - Caspofungin: an advanced treatment approach for suspected or confirmed invasive aspergillosis. AB - Invasive aspergillosis occurs in a wide range of immunocompromised patients and is typically associated with an extremely poor prognosis. Caspofungin represents an additional therapeutic alternative to standard antifungal therapies in patients with suspected or confirmed invasive aspergillosis, as evidenced by a growing body of experience confirming its utility in treating this patient population. Caspofungin has demonstrated clinical efficacy when administered as salvage therapy in patients refractory to or intolerant of standard antifungal therapies, as first-line empirical therapy in patients with persistent febrile neutropenia and as combination therapy in difficult-to-treat patients refractory to or intolerant of standard therapies. Further studies are warranted to establish the effectiveness of caspofungin either alone or in combination as primary therapy for invasive aspergillosis. PMID- 16701981 TI - Managing the challenges of invasive fungal infections. PMID- 16701982 TI - Antifungal drug resistance: limited data, dramatic impact? AB - As the clinical use of antifungal agents continues to grow, the spectre of antifungal drug resistance inevitably emerges. This has been most evident with the extensive prescription of fluconazole in HIV-positive patients. In other settings, there has also been concern over changing patterns in the epidemiology of Candida infections related to fluconazole use. There have been some observations on how new antifungals such as voriconazole and caspofungin are impacting the shift in pathogens and drug resistance. For lipid formulations of amphotericin B, drug resistance is uncommon. Although antifungal drug susceptibility tests for Candida spp. are becoming increasingly standardized, there are limited data that confirm a link between in vitro resistance and treatment failure. Hence, more evidence-based data are needed. This paper examines some of the data on antifungal resistance and briefly explores suitable clinical management strategies for treating fungal infections. PMID- 16701983 TI - Emergence and prevalence of beta-lactamase-producing Klebsiella pneumoniae resistant to cephems in Japan. AB - Forty-six cephem-resistant Klebsiella pneumoniae strains with minimum inhibitory concentrations>8 microg/mL for cefpodoxime and cefmetazole were selected from clinical isolates obtained between 2000 and 2002 from eight hospitals on Northern Kyushu Island, Japan. We investigated the mechanisms of resistance to cephems in these 46 K. pneumoniae isolates. The results of isoelectric focusing of beta lactamases produced by these isolates, polymerase chain reaction for detection of various Class A, Class B and Class C beta-lactamases, and determination of the sequence of the beta-lactamase structural gene showed that most of these isolates had various types of broad-spectrum beta-lactamases. Of the 46 isolates, 2 were CMY-2 beta-lactamase producers and 41 were DHA-1 beta-lactamase producers. Forty of the 41 DHA-1 beta-lactamase producers simultaneously produced SHV-12 extended spectrum beta-lactamase (ESBL), and the remaining isolate simultaneously produced SHV-27. Furthermore, one DHA-1 and SHV-12 beta-lactamase producer also produced IMP-1 beta-lactamase. The only broad-spectrum beta-lactamase with another isolate was IMP-1. Chromosomal DNA restriction fragment analysis using XbaI suggested that nosocomial infection due to DHA-1 and SHV-12 beta-lactamase producers had occurred at two centres. This is the first report of nosocomial infection due to DHA-1 beta-lactamase-producing K. pneumoniae including other plasmid-encoded AmpC beta-lactamases in Japan. The mechanisms of resistance of 44 of the 46 isolates to cephalosporins and cephamycins were ESBL production and/or plasmid-encoded AmpC beta-lactamase and/or IMP-1 beta-lactamase production. For two isolates, the mechanism of resistance to could not be identified. These results show that it is necessary to minimise the prevalence of these resistant strains as it will be a very serious problem if organisms producing these broad-spectrum beta-lactamases increase in clinical situations. It is important to detect these strains sooner and to perform rigorous infection control earlier. PMID- 16701984 TI - Mechanisms underlying long- and short-range nodal signaling in Zebrafish. AB - Precise regulation of the signaling range of secreted molecules is essential for proper pattern formation during development. The Nodal family of TGF-beta proteins has been shown to function as both short- and long-range signals. But the underlying mechanisms remain elusive. In this study, we investigated the regulation of the signaling range of zebrafish Nodal proteins Cyclops and Squint, which are short- and long-range signals, respectively. We show that (1) the stability of Cyclops and Squint correlates with the activity range but increasing the stability of the short-range Cyclops does not increase its signaling range; (2) structural differences in the N-terminus region of the mature peptides of Cyclops and Squint determine their differences in the signaling range and swapping the N-terminus region of the Squint mature ligand into that of Cyclops makes the latter function at a distance. PMID- 16701985 TI - Zebrafish Numb homologue: phylogenetic evolution and involvement in regulation of left-right asymmetry. AB - Numb and its homologue, Numb-like (Numbl), play important roles in mammalian development, but their role in embryonic development of lower vertebrates remains unknown. We cloned a zebrafish numb homologue (znumb) by searching database. znumb shows approximately 60% identity with mammalian Numb orthologs. Interstingly, znumb lacks two specific sequence motifs unique to Numbl orthologs. However, chromosomal localization of znumb gene revealed colinearity with genes located around mouse and human Numbl genes. Furthermore, multi-species comparisons of conserved phosphotyrosine-binding (PTB) domain sequences in Numb and Numbl proteins suggest that znumb is more closely related to Numbl than Numb. znumb mRNA was expressed in a wide variety of zebrafish adult tissues. Overexpression of znumb in embryos resulted in an absence, or reversal, of the normal leftward shift of the developing heart tube. Furthermore, no or bi-lateral transcripts of lefty2 were observed in znumb-expressing embryos, suggesting that the Notch signaling was essential for left-right field formation and maintenance in zebrafish, and that znumb perturbed this process through down-regulation of endogenous Notch signaling. PMID- 16701987 TI - Every new system generates new problem a priori. PMID- 16701986 TI - Deformation and nano-rheology of red blood cells: an AFM investigation. AB - Interaction forces, deformation and nano-rheology of individual red blood cells in physiologically relevant solution conditions have been determined by colloid probe atomic force microscopy (AFM). On approach of the physically immobilised cell and silica glass spherical probe surfaces, deformation of the red blood cell was observed in the force curves. At low levels of deformation, spring constants were determined in the range 3-6 m Nm(-1), whereas for higher levels of deformation, the forces increase non-linearly and on retraction, significant force curve hysteresis is observed (i.e. lower forces upon retraction). The extent of force curve hysteresis was dependent on both the drive velocity and loading force, typical of a viscoelastic system. The response of the red blood cell has been described by viscoelastic theory, where the short and long time scale elastic moduli and relaxation times are determined, i.e. the cell's nano rheological properties elucidated. In addition to a time independent elastic modulus of 4.0 x 10(3)Nm(-2) at low levels of deformation, time-dependent elastic moduli ranges are observed (3.5 x 10(4) to 5.5 x 10(4)Nm(-2) at intermediate levels of deformation and 1.5 x 10(5) to 3.0 x 10(5)Nm(-2) at higher levels of deformation). That is, one elastic and more than one viscoelastic response to the red blood cell deformation is evident, which is considered to reflect the cellular structure. PMID- 16701988 TI - Hot-tub lung: hypersensitivity to Mycobacterium avium but not hypersensitivity pneumonitis. PMID- 16701989 TI - Once-daily dosing with budesonide/formoterol compared with twice-daily budesonide/formoterol and once-daily budesonide in adults with mild to moderate asthma. AB - Adherence to maintenance therapy is often poor in patients with asthma. Simplifying dosing regimens has the potential to improve both adherence and asthma-related morbidity. In this 12-week, randomized, double-blind, double dummy, parallel-group study, 617 patients with mild to moderate persistent asthma (mean forced expiratory volume in 1s [FEV1] 78.5% predicted) who were not optimally controlled on inhaled corticosteroids (200-500 microg/day) were randomized to once-daily budesonide/formoterol (80/4.5 microg, 2 inhalations in the evening), twice-daily budesonide/formoterol (80/4.5 microg, 1 inhalation), or a corresponding dose of budesonide once-daily (200 microg, 1 inhalation in the evening). All patients received budesonide (100 microg twice daily) during a 2 week run-in. Changes in mean morning peak expiratory flow (PEF) were similar for od budesonide/formoterol (23.4 l/min) and twice-daily budesonide/formoterol (24.1 l/min), and both were greater than with budesonide (5.5 l/min; both P<0.001). Evening PEF, symptom-free days, reliever-free days, and asthma control days were improved with budesonide/formoterol therapy vs. budesonide (P<0.05 vs. budesonide for all variables). All treatments were well tolerated. Budesonide/formoterol administered once daily in the evening is a convenient treatment regimen that is as effective in improving asthma control as twice-daily dosing in patients with mild to moderate persistent asthma. PMID- 16701990 TI - A fluorescence enhancement-based sensor using glycosylated metalloporphyrin as a recognition element for levamisole assay. AB - A fluorescence sensor based on the supermolecular recognition by glycosylated metalloporphyrin for levamisole (LEV) assay is reported. For the preparation of a LEV-sensitive active material, 5, 10, 15, 20-tetrakis[2-(2, 3, 4, 6-tetraacetyl beta-D-glucopyranosyl)-1-O-phenyl] porphyrin and its metal complexes were synthesized and used in an optode membrane prepared by including glycosylated metalloporphyrin in chitosan matrice. The immobilized glycosylated metalloporphyrin is shown to be weakly fluorescent as a result of the inhibiting of the electron tansfer by central metal. The fluorescence enhancement of the metalloporphyrin modified optode membrane by LEV is based on the complexation with the central metal moiety of metalloporphyrin and weakening the inhibiting of the electron tansfer for metalloporphyrin. The glycosylated metalloporphyrin/chitosan optode membrane showed excellent selectivity toward LEV with respect to a number of interferents and exhibited stable response. The calibration graph obtained with the proposed sensor was linear over the range of 1.3x10(-5)-3.5x10(-7)ML(-1), with a detection limit of 3.5x10(-7)ML(-1) for LEV. The prepared sensor is applied for the determination of LEV in pharmaceutical preparations and the results agreed with the values obtained by the pharmacopoeia method. PMID- 16701991 TI - Plant genetic engineering to improve biomass characteristics for biofuels. AB - Currently, most ethanol produced in the United States is derived from maize kernel, at levels in excess of four billion gallons per year. Plant lignocellulosic biomass is renewable, cheap and globally available at 10-50 billion tons per year. At present, plant biomass is converted to fermentable sugars for the production of biofuels using pretreatment processes that disrupt the lignocellulose and remove the lignin, thus allowing the access of microbial enzymes for cellulose deconstruction. Both the pretreatments and the production of enzymes in microbial tanks are expensive. Recent advances in plant genetic engineering could reduce biomass conversion costs by developing crop varieties with less lignin, crops that self-produce cellulase enzymes for cellulose degradation and ligninase enzymes for lignin degradation, or plants that have increased cellulose or an overall biomass yield. PMID- 16701992 TI - Microbial ecology of drinking water distribution systems. AB - The supply of clean drinking water is a major, and relatively recent, public health milestone. Control of microbial growth in drinking water distribution systems, often achieved through the addition of disinfectants, is essential to limiting waterborne illness, particularly in immunocompromised subpopulations. Recent inquiries into the microbial ecology of distribution systems have found that pathogen resistance to chlorination is affected by microbial community diversity and interspecies relationships. Research indicates that multispecies biofilms are generally more resistant to disinfection than single-species biofilms. Other recent findings are the increased survival of the bacterial pathogen Legionella pneumophila when present inside its protozoan host Hartmannella vermiformis and the depletion of chloramine disinfectant residuals by nitrifying bacteria, leading to increased overall microbial growth. Interactions such as these are unaccounted for in current disinfection models. An understanding of the microbial ecology of distribution systems is necessary to design innovative and effective control strategies that will ensure safe and high quality drinking water. PMID- 16701993 TI - New opportunities revealed by biotechnological explorations of extremophiles. AB - Over the past few decades the extremes at which life thrives has continued to challenge our understanding of biochemistry, biology and evolution. As more new extremophiles are brought into laboratory culture, they have provided a multitude of potential applications for biotechnology. More recently, innovative culturing approaches, environmental genome sequencing and whole genome sequencing have provided new opportunities for the biotechnological exploration of extremophiles. PMID- 16701994 TI - Capturing the uncultivated majority. AB - The metagenomic analysis of environmental microbial communities continues to be a rapidly developing area of study. DNA isolation, the first step in capturing the uncultivated majority, has seen many advances in recent years. Protocols have been developed to distinguish DNA from live versus dead cells and to separate extracellular from intracellular DNA. Looking to increase our understanding of the role that members of a microbial community play in ecological processes, several techniques have been developed that are enabling greater in-depth analysis of environmental metagenomes. These include the development of environmental gene tags and the serial analysis of 16S rRNA gene sequence tags. In addition, new screening methods have been designed to select for specific functional genes within metagenomic libraries. Finally, new cultivation methods continue to be developed to improve our ability to capture a greater diversity of microorganisms within the environment. PMID- 16701995 TI - POMT2 mutation in a patient with 'MEB-like' phenotype. AB - Mutations in POMT2 have so far only been reported in patients with Walker-Warburg phenotype. We report heterozygous POMT2 mutations in an a girl with a milder phenotype characterized by mental retardation, microcephaly, hypertrophy of the quadriceps and calf muscles, and structural brain changes mostly affecting the posterior fossa. Our findings suggest that, as previously reported for POMT1 and FKRP, mutations in the POMT2 can also be associated with clinical heterogeneity. PMID- 16701996 TI - 135th ENMC International Workshop: nutrition in amyotrophic lateral sclerosis 18 20 of March 2005, Naarden, The Netherlands. PMID- 16701997 TI - The robustness of age-related gait adaptations: can running counterbalance the consequences of ageing? AB - Previous studies showed age-related redistribution of joint torques from ankle joint plantar flexion to hip joint extension in gait. In the present study it was hypothesized that running can prevent the occurrence of this joint torque redistribution. Four groups of subjects participated in this study (young and elderly both physically active and inactive). All subjects walked at a comfortable, preferred velocity and at an imposed velocity of 1.5m/s. Kinematics of lower limb segments and ground reaction forces were assessed. Inverse dynamics method was applied to determine torques around ankle, knee and hip joints. A redistribution of joint torques from plantar flexion to hip joint extension was found to occur in both active and inactive elderly. However, the active elderly had a larger increase of the hip extension torque. By this they are able to maintain the support torque at the level of young subjects. Inactive elderly displayed reduced support torques. It is concluded that the age-related redistribution of joint torques is an important phenomenon. Frequent running does not prevent this shift. Active elderly increase this redistribution to compensate for muscle function reduction. PMID- 16701998 TI - Thermally stimulated luminescence and EPR studies on topaz. AB - In the present work, electron paramagnetic resonance (EPR) spectra and thermoluminescence (TL) emission of colourless natural topaz from Santo Antonio do Jacinto, Brazil, was studied as a function of thermal treatment and gamma irradiation dose, focussing on the use of this material as a radiation dosimeter. EPR measurements on "as-received" samples at room temperature provided signals in the region of about 500 at 5,000 G. The signal located around g approximately equal to 2, frequently attributed to (AlO(4))(0), increased with additional gamma dose and disappeared after thermal treatment at 500 degrees C for 1h. Irradiation after thermal treatment recovered this signal. The decay promoted by isochronal thermal treatment showed the (AlO(4))(0) defects to be directly related to the TL glows peaks. The variation of EPR spectrum with annealing temperature prior to irradiation showed that the variation of TL sensitivity is a consequence of the variation of the (AlO(4))(0) population. PMID- 16701999 TI - Effects of sparfloxacin on CNS functions and urinary hydroxyproline in mice. AB - Sparfloxacin is a widely prescribed drug for various infections. In the present study, sparfloxacin, at two doses, 25 and 50 mg kg(-1) was screened on seven neurological and neurobehavioural parameters in mice. In addition, a chronic study was performed to measure the grip strength of the animal to simulate tendon damage and corresponding hydroxyproline levels were determined in mice urine. Increasing current electroshock test (ICES), pentylenetetrazole (PTZ)-induced seizures, forced swim test (FST), passive avoidance response (PAR), spontaneous alternation behaviour (SAB) and elevated plus maze (EPM) were used for studying the acute effects of sparfloxacin while ICES, grip strength test (GST) and urinary hydroxyproline estimations were done for studying its chronic effects. The results indicate a proconvulsive potential of sparfloxacin on the ICES test (when administered for 21 days) and on PTZ when administered for 4 days. Further sparfloxacin (50 mg kg(-1)x4 days) exhibited anxiety like effects on EPM while mood and memory remained unaffected. Sparfloxacin 50 mg kg(-1) reduced the grip strength of mice after 2 weeks while both 25 and 50 mg kg(-1) reduced the same after 3 weeks. On urinary hydroxyproline, sparfloxacin 25 mg kg(-1) significantly increased the levels after 3 weeks of treatment. At 50 mg kg(-1), an elevation was evident from the first week onwards. In conclusion, sparfloxacin produced proconvulsant and anxiogenic effects in mice and was devoid of effects on mood and memory. On chronic treatment, it reduced grip strength and caused an elevation in urinary hydroxyproline levels. PMID- 16702000 TI - Effects of lipopolysaccharide on gene expression of antimicrobial peptides (penaeidins and crustin), serine proteinase and prophenoloxidase in haemocytes of the Pacific white shrimp, Litopenaeus vannamei. AB - For shrimp immune defences, prophenoloxidase (proPO) activating system and antimicrobial peptides in circulating haemocytes play important roles. In the present study, the effects of lipopolysaccharide (LPS) injection on gene expression of penaeidins, crustin, serine proteinase and proPO in haemocytes were determined using real-time reverse transcription-polymerase chain reaction (PCR) in the Pacific white shrimp Litopenaeus vannamei. After injection of LPS, mRNA levels of antimicrobial peptides, penaeidin 2 (PEN2), penaeidin 3 (PEN3), penaeidin 4 (PEN4) and crustin decreased in a dose-dependent manner, while mRNA levels of serine proteinase and proPO did not change significantly. In a time course experiment, injection of LPS caused significant depression in mRNA levels of PEN2, PEN3, PEN4 and crustin at 4h post-injection, and the depressed mRNA levels returned to initial levels by 72h post-injection. On the other hand, mRNA levels of serine proteinase and proPO did not show a significant change after injection. These results suggest that the antimicrobial peptide system responds to LPS injection at a gene expression level while the proPO system does not respond at a gene expression level. PMID- 16702001 TI - Voxel-based morphometry in patients with idiopathic generalized epilepsies. AB - Idiopathic generalized epilepsies (IGE) are a group of frequent age-related epilepsy syndromes. IGE are clinically characterized by generalized tonic-clonic, myoclonic and absence seizures. According to predominant seizure type and age of onset, IGE are divided in subsyndromes: childhood absence and juvenile absence epilepsy (AE), juvenile myoclonic epilepsy (JME) and generalized tonic-clonic seizures on awakening (GTCS). The limits between these subsyndromes are not well defined, supporting the existence of only one major syndrome. Visual assessment of routine magnetic resonance imaging (MRI) in patients with IGE is normal. MRI voxel-based morphometry (VBM) uses automatically segmented gray and white matter for comparisons, eliminating the investigator bias. We used VBM to study 120 individuals (47 controls, 44 with JME, 24 with AE and 15 with GTCS) to investigate the presence of subtle structural abnormalities in IGE subsyndromes. VBM was performed searching for abnormalities on gray matter concentration (GMC) between patients groups and controls. Compared to controls, JME presented increased GMC in frontobasal region and AE showed increased GMC in the superior mesiofrontal region. The GTCS group did not differ from controls. There were no areas of reduced GMC with the statistical level selected. Region of interest analysis showed increased GMC in the anterior portion of the thalamus in patients with absence seizures. Our results support subtle GMC abnormalities in patients with JME and AE when compared to controls. These findings suggest the existence of different patterns of cortical abnormalities in IGE subsyndromes. PMID- 16702002 TI - The left intraparietal sulcus and verbal short-term memory: focus of attention or serial order? AB - One of the most consistently activated regions during verbal short-term memory (STM) tasks is the left intraparietal sulcus (IPS). However, its precise role remains a matter of debate. While some authors consider the IPS to be a specific store for serial order information, other data suggest that it serves a more general function of attentional focalization. In the current fMRI experiment, we investigated these two hypotheses by presenting different verbal STM conditions that probed recognition for word identity or word order and by assessing functional connectivity of the left IPS with distant brain areas. If the IPS has a role of attentional focalization, then it should be involved in both order and item conditions, but it should be connected to different brain regions, depending on the neural substrates involved in processing the different types of information (order versus phonological/orthographic) to be remembered in the item and order STM conditions. We observed that the left IPS was activated in both order and item STM conditions but for different reasons: during order STM, the left IPS was functionally connected to serial/temporal order processing areas in the right IPS, premotor and cerebellar cortices, while during item STM, the left IPS was connected to phonological and orthographic processing areas in the superior temporal and fusiform gyri. Our data support a position considering that the left IPS acts as an attentional modulator of distant neural networks which themselves are specialized in processing order or language representations. More generally, they strengthen attention-based accounts of verbal STM. PMID- 16702003 TI - Bistable illusory rebound motion: Event-related functional magnetic resonance imaging of perceptual states and switches. AB - The neural correlates of a recently discovered visual illusion that we call 'illusory rebound motion' (IRM) are described. This illusion is remarkable because motion is perceived in the absence of any net motion energy in the stimulus. When viewing bars alternating between white and black on a gray background, the percept alternates between one of flashing bars (veridical) and the IRM illusion, where the bars appear to shoot back and forth rather like the opening and closing of a zipper. The event-related functional magnetic resonance imaging (fMRI) data reported here reveal that (1) the blood-oxygen-level dependent (BOLD) signal in the human analog of macaque motion processing area MT (hMT+) increases when there is a perceptual change from "no-IRM" to "see-IRM" and decreases when there is a perceptual change from "see-IRM" to "no-IRM," although the stimulus remains constant; and (2) the BOLD signal in early retinotopic areas (V1, V2, and V3d) shows switch-related activation whenever there is a perceptual change, regardless whether from IRM to no-IRM or vice versa. We conclude that hMT+ is a neural correlate of this novel illusory motion percept because BOLD signal in hMT+ modulates with the perception of IRM. PMID- 16702004 TI - Cardiopulmonary interaction in heart failure. AB - In heart failure lung dysfunction is frequent and is greater the greater the heart failure severity. It can be evaluated in terms of lung mechanics and gas diffusion. Indeed heart-lung interaction is related to heart dimensions and lung fluid content; furthermore heart-lung interaction is influenced by the body position. Lung diffusion is also altered in patients with chronic heart failure, and a low gas diffusion is associated with a reduced performance. During exercise, heart-lung interaction becomes more evident. Heart failure patients show an abnormal hyperventilation due to a progressively increased respiratory rate, and a lower tidal volume; hyperventilation is due to different causes including enhanced responses from chemo- and metabolo-receptors, increased CO(2) production and increased dead space ventilation. Several drugs affect the ventilatory pattern in heart failure patients: ACE-inhibitors and anti aldosteronic drugs improve lung diffusion and ventilatory efficiency during exercise; beta-blockers reduce exercise-induced hyperventilation. Furthermore, ultrafiltration improves lung mechanics, both at rest and during exercise, through body fluid content reduction. PMID- 16702005 TI - Circadian orchestration of developmental hormones in the insect, Rhodnius prolixus. AB - This review presents a new perspective on the circadian regulation and functions of insect developmental hormones. In Rhodnius prolixus (Hemiptera), the brain neuropeptide prothoracicotropic hormone (PTTH) is released with a circadian rhythm that is controlled by paired photosensitive clocks in the brain. These clocks comprise the dorsal and lateral PER/TIM clock neurons known to regulate behavioral rhythms in Drosophila. Axons of PTTH and clock cells make close contact. Photosensitive PER/TIM clocks also reside in the paired prothoracic glands (PGs), which generate rhythmic synthesis and release of the ecdysteroid molting hormones. The PG clocks are entrained by both light and PTTH. These four clocks are coupled together by both nerves and hormones into a timing system whose primary regulated output is the circadian rhythm of ecdysteroids in the hemolymph. This complex timing system appears necessary to ensure circadian organization of the gene expression that is induced in target cells by ecdysteroids via circadian cycling of the nuclear ecdysteroid receptor (EcR). This multioscillator system serves to transduce 'the day outside' into endocrine rhythms that orchestrate 'the day inside'. It has many functional similarities with vertebrate circadian systems. PMID- 16702006 TI - Efficacy and safety of growth hormone (GH) in the treatment of adult Japanese patients with GH deficiency: a randomised, placebo-controlled study. AB - OBJECTIVE: The aim of this study was to assess the effect of growth hormone (GH) replacement therapy on lean body mass (LBM) and other variables including body fat mass, serum lipids and quality of life measures in GH-deficient Japanese adults. DESIGN: This was a multicentre, double-blind, placebo-controlled, parallel group study. Following initial screening, patients were randomly assigned to GH treatment (n=37) or placebo (n=36). GH treatment was started at an initial dose 0.003 mg/kg/day s.c. each day for the first 4 weeks after which the dose was increased to 0.006 mg/kg/day for 4 weeks and then to 0.012 mg/kg/day for the last 16 weeks (n=37). Body composition, serum lipids, serum IGF-I and IGFBP-3 levels were measured during the 24-week study. Short Form-36 and Quality of Life Assessment of GH Deficiency in Adults scores were also determined. RESULTS: LBM was significantly increased from baseline at 24 weeks in GH-treated patients, with a mean (+/-SD) increase of 4.7% (+/-5.3%) compared with an increase of 1.0% (+/-4.4%) in the placebo group (p<0.0001 versus baseline, p=0.0003 versus placebo). Percentage body fat decreased significantly from baseline in GH-treated patients (9.3%, p<0.0001), compared with a non-significant 0.2% increase in the placebo group (p<0.0004 for difference between treatment groups). In addition, significantly increased serum IGF-I and IGFBP-3 levels and improvements in the patients' serum lipid profiles were observed in patients who received GH therapy. Changes in quality of life measures did not differ between treatments, probably because of the small number of patients studied. GH therapy was well tolerated, with adverse events of any cause reported in 86.5% of the GH treatment group and 83.3% of the placebo group. CONCLUSION: GH treatment significantly improved body composition and serum lipid profiles in adult Japanese patients with GH deficiency compared with placebo and had no clinically relevant adverse effects. PMID- 16702007 TI - Extended-spectrum beta-lactamases (ESBL) in community isolates from North India: frequency and predisposing factors. PMID- 16702008 TI - Recurrent catheter-related bloodstream infections: Risk factors and outcome. AB - OBJECTIVE: To identify risk factors for recurrent catheter related bloodstream infections (CR-BSIs). The study was undertaken at the University of Utah Hospital and involved patients who had a CR-BSI followed by catheter removal and reinsertion between January 1998 and February 2002. DESIGN: A retrospective chart review for the cohort study of catheters initially infected, which were then followed to study risk factors for a subsequent infection. Both central line and peripherally inserted central line catheters were included in the study. A recurrent CR-BSI was defined as positive blood cultures after three negative cultures, coupled with positive catheter tip culture or no other evident new source of infection. RESULTS: Twenty-five (34%) of 73 patients had a recurrent CR BSI. The first CR-BSI occurred a mean of 20.4 days after catheter insertion whereas recurrence developed a mean of 12.1 days after reinsertion (p = 0.392). Coagulase-negative staphylococci (60%) were the most common cause of recurrent infection. The recurrence was more common among the patients who were given blood product transfusion (hazard ratio (HR) 2.3; confidence interval (CI) 1.02-5.67, p = 0.049). In 20 (27%) patients, catheters were changed over a guidewire. The guidewire catheter exchange was not found to be associated with an increased risk of recurrent infection (p = 0.582). CONCLUSION: Catheter replacement to a new site, instead of rewiring, was not shown to decrease the risk for recurrent infection. The transfusion of blood products was associated with an increased risk for recurrent infection. PMID- 16702009 TI - Drug-induced death of the asexual blood stages of Plasmodium falciparum occurs without typical signs of apoptosis. AB - There is clear evidence that most antimalarial drugs, while acting through different mechanisms, are associated with parasite growth/development inhibition and eventual parasite death. However, the exact mode of parasite death remains unclear. In the present study, we investigated the ability of various drugs, including two antimalarial drugs (chloroquine and atovaquone), a topoisemerase II inhibitor (etoposide) and a nitric oxide donor (S-nitro-N-acetyl-D, L penicillamine), to induce apoptosis in a laboratory strain of Plasmodium falciparum. Results obtained from flow cytometric analysis showed a significant reduction in the percent of parasitemia and parasite growth in all drug-treated parasite cultures, including those treated with etoposide and S-nitro-N-acetyl-D, L-penicillamine. For further investigation, we used various biochemical approaches including the terminal dUTP nick-end labeling assay, determination of mitochondrial membrane integrity and DNA degradation/fragmentation, to analyze the changes occurring during parasite-drug interactions and eventual death. We observed that loss of membrane potential was induced in parasite cultures treated with atovaquone, while S-nitro-N-acetyl-D, L-penicillamine induced abnormal parasite forms, "crisis forms", and minor DNA degradation. However, these features were not observed in the parasite cultures treated with chloroquine nor were other features of apoptosis-like death associated with any of the drugs used in this study. The death resulting from the various drug treatments is atypical of apotosis. More studies will be needed to define the precise mode of death exhibited by P. falciparum. PMID- 16702011 TI - Impaired T-cell differentiation in the thymus at the early stages of acute pathogenic chimeric simian-human immunodeficiency virus (SHIV) infection in contrast to less pathogenic SHIV infection. AB - One of the mechanisms by which HIV infection induces the depletion of CD4+ T cells has been suggested to be impairment of T-cell development in the thymus, although there is no direct evidence that this occurs. To examine this possibility, we compared T-cell maturation in the intrathymic progenitors between macaques infected with an acute pathogenic chimeric simian-human immunodeficiency virus (SHIV), which causes profound and irreversible CD4+ T-cell depletion, and macaques infected with a less pathogenic SHIV, which causes only a transient CD4+ T-cell decline. Within 27 days post-inoculation (dpi), the two virus infections caused similar increases in plasma viral loads and similar decreases in CD4+ T cell counts. However, in the thymus, the acute pathogenic SHIV resulted in increased thymic involution, atrophy and the depletion of immature T cells including CD4(+)CD8(+) double-positive (DP) cells, whereas the less pathogenic SHIV did not have these effects. Ex vivo differentiation of CD3(-)CD4(-)CD8(-) triple-negative (TN) intrathymic progenitors to DP cells was assessed by a monkey mouse xenogenic fetal thymus organ culture system. Differentiation was impaired in the TN intrathymic progenitors of the acute pathogenic SHIV-infected monkeys, while differentiation was not impaired in the TN intrathymic progenitors of the less pathogenic SHIV-infected monkeys. These differences suggest that dysfunction of thymic maturation makes an important contribution to the irreversible depletion of circulating CD4+ T cells in vivo. PMID- 16702010 TI - Induction of human immunodeficiency virus neutralizing antibodies using fusion complexes. AB - Human immunodeficiency virus-1 (HIV-1) infects cells by membrane fusion that is mediated by the envelope proteins gp120/gp41 and the cellular receptors CD4 and CCR5. During this process, some conserved viral epitopes are temporarily exposed and may induce a neutralizing antibody response when fixed in the fusogenic conformation. These transient structures are conserved and may be effective antigens for use in an anti-HIV-1 vaccine. In this study we tested different conditions of preparation of fusion complexes inducing neutralizing antibodies against both R5 and X4 tropic HIV-1 strains. Cell lines expressing HIV-1 gp120/gp41 and CD4-CCR5 were prepared and conditions for producing fusion complexes were tested. Complexes produced at different temperature and fixative combinations were used to immunize mice. Results indicated that (a) fusion complexes prepared at either 21 degrees C, 30 degrees C or 37 degrees C were immunogenic and induced neutralizing antibodies against both R5 and X4 HIV-1 heterologous isolates; (b) after extensive purification of antibodies there was no cytotoxic effect; (c) complexes prepared at 37 degrees C were more immunogenic and induced higher titers of neutralizing antibodies than complexes prepared at either 21 degrees C or 30 degrees C; (d) the fixative used did not affect the titer of neutralizing antibodies except for glutaraldehyde which was ineffective; (e) the neutralizing activity was retained after CD4-CCR5 antibody removal. The production of higher titers of neutralizing antibody with fusion complexes prepared at 37 degrees C, as compared to lower temperatures, may be related to the induction of antibodies against many different conformation intermediates that subsequently act synergistically at different steps in the fusion process. PMID- 16702012 TI - Induction of polymorphonuclear leukocyte response by human cytomegalovirus. AB - Neutrophils are important in the defense against bacterial infections, by ingesting and killing invading microorganisms. Because of the higher incidence of bacterial infections in patients with active human cytomegalovirus (HCMV) infections, we hypothesized that HCMV-infected neutrophils were inefficient in eliminating the bacteria. Therefore, we mock infected or infected neutrophils with HCMV by contact with HCMV-infected human pulmonary artery endothelial cells. We found that HCMV infection without N-formyl-methionyl-leucyl-phenylalanine (fMLP) stimulation increased the surface expression of CD11b to the same extent as fMLP stimulation of mock infected cells. Also, HCMV-infected neutrophils became more efficient in phagocytosing serum opsonized yeast particles than mock infected cells. Furthermore, we observed an increase in intracellular free calcium and chemiluminescence in HCMV-infected cells, in response to fMLP compared to fMLP-treated mock cells. We also found that apoptosis was significantly inhibited in HCMV-infected neutrophils. In conclusion, our results suggest that neutrophils become more effective in performing their effector functions when infected with HCMV. Thus, the higher incidence of bacterial infections in HCMV patients might not be due directly to a dysfunction in the neutrophils. Instead, the fact that apoptosis is inhibited may cause over reactive neutrophils to remain in the tissues, where they will start leaking their contents, damaging the tissues and contributing to inflammatory processes. PMID- 16702013 TI - Streptococcus pyogenes induces epithelial inflammatory responses through NF kappaB/MAPK signaling pathways. AB - Innate immunity involves a cascade of inflammatory events, resulting in the secretion of chemokines and cytokines to recruit mediator cells in adaptive immunity. To study epithelial inflammatory responses initiated by Streptococcus pyogenes infection, we investigated chemotaxis ability in the supernatant of infected human respiratory epithelial HEp-2 cells. Our results showed that these supernatants showed significantly increased ability to attract monocytes, implying the release of inflammatory chemoattractants into the medium. Expression of interleukin (IL)-8 and IL-6 in HEp-2 cells was significantly increased at both the mRNA and protein levels after infection with S. pyogenes. Electrophoretic mobility shift and reporter-gene assays demonstrated that the transcription factors NF-kappaB and AP-1, regulated by mitogen-activated protein (MAP) kinase, were activated after streptococcal infection. The increases in mRNAs for IL-8 and IL-6 were abrogated by addition of NF-kappaB and MAP kinase inhibitors, suggesting that the upregulation of IL-8 and IL-6 is mediated through NF-kappaB and MAP kinase signaling pathways. Taken together, our results indicate that S. pyogenes infection of epithelial cells induces the secretion of pro-inflammatory chemokines/cytokines through activation of NF-kappaB and MAP kinase signaling pathways. These early innate responses initiated by S. pyogenes-infected respiratory epithelial cells may recruit immune cells to the airway and induce inflammation. PMID- 16702014 TI - Salmonella carrier-state in hens: study of host resistance by a gene expression approach. AB - Salmonellosis is one of the main causes of food-borne poisoning due to the consumption of contaminated poultry products. In the flocks, Salmonella is able to persist in the digestive tract of birds for weeks without triggering any symptom. In order to identify molecules and genes involved in the mechanism of host resistance to intestinal carrier-state, two different inbred lines of laying hens were orally inoculated with Salmonella Enteritidis. Bacterial colonization and host gene expression were measured in the caecum and its sentinel lymphoid tissue, respectively. Significantly increased expression of chemokine, anti infectious cytokine, bacterial receptor, antimicrobial mediator and particularly, defensin genes was observed in the line carrying a lower level of bacteria in the caecum. These innate immunity molecules were either constitutively or inductively highly expressed in resistant adult birds and thus present candidate genes to play an important role in the host defence against Salmonella colonization. PMID- 16702015 TI - Is the reactive oxygen species-dependent-NF-kappaB activation observed in iron loaded BALB/c mice a key process preventing growth of Leishmania major progeny and tissue-damage? AB - Systemic iron delivery to BALB/c mice, at time points surrounding the inoculation of 1000 Leishmania major metacyclic promastigotes intradermally in the ear results in the complete absence of onset and further development of ear lesion. In these iron-protected mice, the L. major intracellular progeny remains very low in both the ear and the draining lymph node. The iron-induced protective status is associated with a diphenyleneiodonium-sensitive sustained increased oxidative burst. We showed that iron-loaded mice developed no lesions at the site of the primary inoculation and were also resistant to reinoculation at a distant site (intradermal re-inoculation of 1000 metacyclic promastigotes in the contra lateral ear). Interestingly, in the lymph node cell population recovered from iron-loaded mice at weeks 8 and 12 after the second parasite inoculation, and whatever the protective status studied--primary or resistant to re-inoculation- three potentially related features were observed: (i) NF-kappaB activation, (ii) enhanced TCR-mediated T lymphocyte proliferation, and (iii) high number of IFN gamma-positive CD4(+)T cells. These results show a putative role of an iron induced reactive oxygen species-dependent activation of NF-kappaB in the development of protective immunity against L. major. PMID- 16702016 TI - Passive serum therapy with polyclonal antibodies against Mycobacterium tuberculosis protects against post-chemotherapy relapse of tuberculosis infection in SCID mice. AB - We investigated the protective role of immune-sera against reactivation of Mycobacterium tuberculosis infection in SCID mice and found that passive immunization with sera obtained from mice treated with detoxified M. tuberculosis extracts (delivered in liposomes in a composition known as RUTI) exerted significant protection. Our SCID mouse model consisted of aerosol infection by M. tuberculosis, followed by 3 to 8weeks of chemotherapy with isoniazid+rifampicin (INH+RIF) (25 and 10mg/kg, respectively). After infection and antibiotic administration, two groups of mice were treated for up to 10weeks with intraperitoneal passive immunization using hyperimmune serum (HS) obtained from mice infected with M. tuberculosis, treated with chemotherapy (INH+RIF) for 8weeks and inoculated with RUTI (HS group) or with normal serum (CT group). Significant differences were found between HS and CT groups in the number of bacilli in the lungs (3.68+/-2.02 vs. 5.72+/-1.41log(10) c.f.u.), extent of pulmonary granulomatomous infiltration (10.33+/-0.67 vs. 31.2+/-1.77%), and percentage of animals without pulmonary abscesses (16.7% vs. 45.5%). These data strongly suggest a protective role of specific antibodies against lung dissemination of M. tuberculosis infection. PMID- 16702018 TI - The evolution from asparagine or threonine to cysteine in position 146 contributes to generation of a more efficient and stable form of muscle creatine kinase in higher vertebrates. AB - Creatine kinase, a key enzyme in vertebrate excitable tissues that require large energy fluxes, catalyzes the reversible transfer of phosphate between adenosine triphosphate and creatine. Sequence alignment indicated that the 146th amino acid is cysteine in the muscle creatine kinase of higher vertebrates including Amphibia, Reptilia, Aves and Mammalia. In fishes, it is cysteine in Agnatha and Chondrichthyes, and asparagine or threonine in Osteichthyes, which is the ancestor of Amphibia, Reptilia, Aves and Mammalia. To explore the structural and functional role of this special residue, a series of site-directed mutants of rabbit muscle creatine kinase were constructed, including C146S, C146N, C146T, C146G, C146A, C146D and C146R. A detailed comparison was made between wild-type creatine kinase and the mutants in catalytic activity, physico-chemical properties and structural stability against thermal inactivation and guanidine hydrochloride denaturation. It was found that except for C146S, the mutants had relatively lower catalytic activity and structural stability than Wt-CK. Wt-CK and C146S were the most stable ones, followed by C146N and C146T, and then C146G and C146A, and C146D and C146R were the least stable mutants. These results suggested that the 146th residue plays a crucial role in maintaining the structural stability of creatine kinase, and that the evolution in this amino acid from asparagine or threonine to cysteine contributes to the generation of a more efficient and more stable form of creatine kinase in higher vertebrates. PMID- 16702017 TI - The subcellular fractionation properties and function of insulin receptor substrate-1 (IRS-1) are independent of cytoskeletal integrity. AB - Efficient insulin action requires spatial and temporal coordination of signaling cascades. The prototypical insulin receptor substrate, IRS-1 plays a central role in insulin signaling. By subcellular fractionation IRS-1 is enriched in a particulate fraction, termed the high speed pellet (HSP), and its redistribution from this fraction is associated with signal attenuation and insulin resistance. Anecdotal evidence suggests the cytoskeleton may underpin the localization of IRS 1 to the HSP. In the present study we have taken a systematic approach to examine whether the cytoskeleton contributes to the subcellular fractionation properties and function of IRS-1. By standard microscopy or immunoprecipitation we were unable to detect evidence to support a specific interaction between IRS-1 and the major cytoskeletal components actin (microfilaments), vimentin (intermediate filaments), and tubulin (microtubules) in 3T3-L1 adipocytes or in CHO.IR.IRS-1 cells. Pharmacological disruption of microfilaments and microtubules, individually or in combination, was without effect on the subcellular distribution of IRS-1 or insulin-stimulated tyrosine phosphorylation in either cell type. Phosphorylation of Akt was modestly reduced (20-35%) in 3T3-L1 adipocytes but not in CHO.IR.IRS-1 cells. In cells lacking intermediate filaments (Vim(-/-)) IRS-1 expression, distribution and insulin-stimulated phosphorylation appeared normal. Even after depolymerisation of microfilaments and microtubules, insulin-stimulated phosphorylation of IRS-1 and Akt were maintained in Vim(-/-) cells. Taken together these data indicate that the characteristic subcellular fractionation properties and function of IRS-1 are unlikely to be mediated by cytoskeletal networks and that proximal insulin signaling does not require an intact cytoskeleton. PMID- 16702019 TI - Suppressing Akt phosphorylation and activating Fas by safrole oxide inhibited angiogenesis and induced vascular endothelial cell apoptosis in the presence of fibroblast growth factor-2 and serum. AB - At present, vascular endothelial cell (VEC) apoptosis induced by deprivation of fibroblast growth factor-2 (FGF-2) and serum has been well studied. But how to trigger VEC apoptosis in the presence of FGF-2 and serum is not well known. To address this question, in this study, the effects of safrole oxide on angiogenesis and VEC growth stimulated by FGF-2 were investigated. The results showed that safrole oxide inhibited angiogenesis and induced VEC apoptosis in the presence of FGF-2 and serum. To understand the possible mechanism of safrole oxide acting, we first examined the phosphorylation of Akt and the activity of nitric oxide synthase (NOS); secondly, we analyzed the expressions and distributions of Fas and P53; then we measured the activity of phosphatidylcholine specific phospholipase C (PC-PLC) in the VECs treated with and without safrole oxide. The results showed that this small molecule obviously suppressed Akt phosphorylation and the activity of NOS, and promoted the expressions of Fas and P53 markedly. Simultaneously, Fas protein clumped on cell membrane, instead of homogenously distributed. The activity of PC-PLC was not changed obviously. The data suggested that safrole oxide effectively inhibited angiogenesis and triggered VEC apoptosis in the presence of FGF-2 and serum, and it might perform its functions by suppressing Akt/NOS signal pathway, upregulating the expressions of Fas and P53 and modifying the distributing pattern of Fas in VEC. This finding provided a powerful chemical probe for promoting VEC apoptosis during angiogenesis stimulated by FGF-2. PMID- 16702020 TI - Caveolin-1 up-regulates CD147 glycosylation and the invasive capability of murine hepatocarcinoma cell lines. AB - CD147 which is a regulator of matrix metalloproteinase (MMP) production on the surface of many malignant tumor cells, shows a highly specific association with caveolin-1 (Cav-1). As a result of heterogeneous N-glycosylation, CD147 exists in both highly glycosylated form, HG-CD147 ( approximately 40-60kDa) and lowly glycosylated form, LG-CD147 ( approximately 32kDa). This study investigated the possible role of Cav-1 in CD147 glycosylation in the HcaF, HcaP and Hepa1-6 mouse hepatocarcinoma cell lines, which have high, low and no metastatic potential in the lymph nodes, respectively, and in the normal mouse liver cell line IAR-20. Using an RNA interference (RNAi) strategy, we showed that the down-regulation of Cav-1 in Hca-F/RNAi cells could suppress the conversion of LG-CD147 to HG-CD147, down-regulate MMP-11 expression and decrease Hca-F/RNAi cell invasion. Conversely, a stable high expression of Cav-1 in Hepa1-6/Cav-1 cell could cause a specific increase of HG-CD147, up-regulate MMP-11 protein expression and enhance Hepa1-6/Cav-1 cell invasion. In conclusion, Cav-1 expression leads to an increased proportion of HG-CD147 relative to LG-CD147, increased production of MMP-11 and a higher invasive capability. Cav-1 is therefore proposed to act as both an oncogene and a tumor suppressor gene, and could represent a new potential target for gene therapy. PMID- 16702021 TI - In situ monitoring of clastogenicity of ambient air in Bratislava, Slovakia using the Tradescantia micronucleus assay and pollen abortion assays. AB - Aim of this study was to monitor the genotoxic effects of polluted air in Bratislava (Slovakia) with the Tradescantia micronucleus (Trad-MN) test. In situ monitoring was carried out at five locations during two seasons (years 2003 and 2004). Flower pots with Tradescantia paludosa (clone 03) plants were exposed for 6-8 weeks at the different sites each year. The highest MN levels were observed in the vicinity of an agrochemical factory (3.1 times higher than background level in 2003 and 2.7 times higher in 2004). Lower effects were seen when plants were exposed to urban traffic emissions or in the vicinity of a glass-producing plant (the MN frequencies ranged between 2.8 and 4.4 per 100 tetrads, respectively, while the control frequencies were 2.1-2.6 per 100 tetrads); exposure near a petrochemical plant had no significant effects. In pollen abortion assays, three wild growing species were used, namely, chicory (Cichorium intybus L.), old man's beard (Clematis vitalba L.) and common toadflax (Linaria vulgaris Mill.). Again, the strongest effects were observed close to the agrochemical industry (reduction of fertile pollen by 5.6%, 11.1% and 8.3% in chicory, old mans beard and in toadflax, respectively). Cichorium intybus was the most sensitive species and the number of abortive pollen grains was 5.1 times higher in specimens collected near the agrochemical factory than that seen at the control location. These observations indicate that contaminated urban air has an impact on the fertility of wild plants. Furthermore, it is interesting that the same rank order of effects was seen in pollen abortion assays as in the Trad-MN test (agrochemical industry>technical glass industry>=traffic>city incinerator/petrochemical plant). These results confirm the sensitivity of the Tradescantia MN test and pollen abortion assays for the detection of air pollution, and show that distinct differences exist in genotoxicity of different sources of pollutants. PMID- 16702022 TI - When requests become orders--a formative investigation into the impact of a computerized physician order entry system on a pathology laboratory service. AB - PURPOSE: The purpose of this study was to identify the key implications of the implementation of a computerized physician order entry (CPOE) system on pathology laboratory services. METHODS: An in-depth qualitative study using observation, focus groups and interviews with pathology staff, managers, clinicians and information systems staff during implementation of a CPOE system in 2004 at a major Australian teaching hospital. RESULTS: Pathology laboratories experienced a shift in their work roles resulting in altered work practices, responsibilities and procedures. These changes were marked by terminological and procedural changes in the test order process from when clinicians issued a request for a test, to the new system that established clinical orders at the point of care. This change was accompanied by some organizational dysfunctions including the emergence of a new category of "frustrated" orders without specimens; problems with the procedure of adding tests to previously existing specimens; the appearance of discrepancies in the recorded time of specimen collection. In response to these changes, hospital and pathology staff adopted a variety of means to cope with their changed circumstances. These ranged from efforts to increase clinical awareness to compensatory laboratory workarounds and enforced rule changes. CONCLUSIONS: CPOE systems can have a major impact on the nature of the work of pathology laboratories. Understanding how and why these changes occur can be enhanced through considering the organizational and social contexts involved. The effectiveness of CPOE systems will rely on how administrators and staff approach and deal with these challenges. PMID- 16702023 TI - Caveolin-1 is required for fatty acid translocase (FAT/CD36) localization and function at the plasma membrane of mouse embryonic fibroblasts. AB - Several lines of evidence suggest that lipid rafts are involved in cellular fatty acid uptake and influence fatty acid translocase (FAT/CD36) function. However, it remains unknown whether caveolae, a specialized raft type, are required for this mechanism. Here, we show that wild-type (WT) mouse embryonic fibroblasts (MEFs) and caveolin-1 knockout (KO) MEFs, which are devoid of caveolae, have comparable overall expression of FAT/CD36 protein but altered subcellular FAT/CD36 localization and function. In WT MEFs, FAT/CD36 was isolated with both lipid raft enriched detergent-resistant membranes (DRMs) and detergent-soluble membranes (DSMs), whereas in cav-1 KO cells it was exclusively associated with DSMs. Subcellular fractionation demonstrated that FAT/CD36 in WT MEFs was localized intracellularly and at the plasma membrane level while in cav-1 KO MEFs it was absent from the plasma membrane. This mistargeting of FAT/CD36 in cav-1 KO cells resulted in reduced fatty acid uptake compared to WT controls. Adenoviral expression of caveolin-1 in KO MEFs induced caveolae formation, redirection of FAT/CD36 to the plasma membrane and rescue of fatty acid uptake. In conclusion, our data provide evidence that caveolin-1 is necessary to target FAT/CD36 to the plasma membrane. Caveolin-1 may influence fatty acid uptake by regulating surface availability of FAT/CD36. PMID- 16702028 TI - Combined therapeutics for insomnia: should our first approach be behavioral or pharmacological? AB - Despite the interest in combining therapeutics many issues remain to be resolved regarding the combination of behavioral treatment and pharmacotherapy for insomnia. Studies examining the relative advantages of cognitive behavioral therapy (CBT) and pharmacotherapy have found that improvements may be achieved more quickly with drug treatment but are more sustained with CBT. Combining modes of treatment is not necessarily superior to monotherapy because long-term effects can vary between patients. Various modes of combination are possible, with one or other type of treatment being started or discontinued depending on the phase of treatment and the patient's response. Which treatment to initiate first, or whether to run treatments concurrently, depends on factors such as the nature of the insomnia, treatment history, comorbid conditions, acceptability of treatment to the patient, and treatment cost or availability. Issues of dosage, treatment duration or whether treatment should be given continuously or intermittently will have an effect on how these treatment types can be integrated to provide the best outcome for the patient. A detailed look at the efficacy of behavioral and pharmacological therapies with regard to different outcome measures gives some indication of how these different types of treatment may act in a complementary fashion; observations that may be exploited in the integration of behavioral with pharmacological approaches in clinical practice. PMID- 16702024 TI - Live cell imaging of phosphoinositide dynamics with fluorescent protein domains. AB - Phosphoinositides make up only a small fraction of membrane phospholipids yet they are of outmost significance in regulating membrane-associated signaling processes. A large number of inositol lipid kinases and phosphatases have evolved to control the rapid production and elimination of these lipids at specific cellular membrane compartments. For a long period of time, the only information about the spatial aspect of inositol lipid metabolism relied upon the immunostaining of enzymes or cell fractionation of the enzyme activities that acted upon these lipids. Recent advances in the understanding of the nature of protein-inositol lipid interactions permitted the design of fluorescent molecular probes that can interact with inositol lipids in a specific manner allowing imaging of phosphoinositide dynamics in live cells. This approach has rapidly gained high popularity, but also provoked criticisms and debate about its limitations. In this review, we will summarize our experience with using these molecular tools and address some issues that most often come up in discussions concerning the usefulness and drawbacks of this technique. The most important value of these debates is that they also challenge our preconceived views of how phosphoinositides regulate cellular functions. PMID- 16702030 TI - The usefulness of artificial neural networks in the evaluation of pulmonary efficiency and antioxidant capacity of welders. AB - OBJECTIVE: The aim of the study was to determine if artificial neural networks (ANNs) may be useful to analyse a complex and large set of data derived from smoking welders for the purpose of finding relationships between parameters describing respiratory system efficiency and antioxidant defence. METHODS: A group of 94 welders employed in a big metallurgic enterprise in Krakow, Poland (men only, aged 29-57 years, all active smokers) occupationally exposed to O(3) and NO(x), were the subjects of this study. They underwent biochemical measurements including total antioxidant status (TAS) and the anti-oxidative defence enzymes superoxide dismutase (SOD) and catalase (CT); biominerals: Fe, Cu, Zn, Mg in blood serum and in hair; the concentrations of albumin, bilirubin, uric acid in blood. The determination of respiratory efficiency was based on a "flow-volume" curve and spirometry. The dependant variables for ANNs were: TAS, SOD, CT. Thirty-one subjects with normal values of all spirometric parameters were selected for the final analysis. RESULTS: Statistically valid relationship between TAS and albumin, Zn and Cu in blood and the two pulmonary parameters forced expiratory volume after 1s (FEV(1)) and middle expiratory flow of 25-75% of vital capacity (MEF(25/75)) were found. Zn concentration almost linearly influenced TAS. For Cu a sigmoid curve was obtained. For albumin concentration as well as for FEV(1) a two-stage curve was observed. CONCLUSIONS: ANNs are useful for the reduction of dimensionality and are suited to analyse a complex and relatively large set of parameters when it is unknown which of these are related. ANNs were useful for finding the relationship between the antioxidant defence and the respiratory system capacity in welders who smoke. PMID- 16702031 TI - Effect of postnatal exposure to caffeine on the pattern of adenosine A1 receptor distribution in respiration-related nuclei of the rat brainstem. AB - Caffeine, which belongs to the methylxantine family of compounds, is commonly ingested in a range of beverages such as coffee, tea, and cola drinks. It is also used therapeutically and is frequently employed in the treatment of respiratory disturbances in human neonates. The aim of the present work has been to examine the ontogeny of the adenosine A1 receptor system in the brainstem of the newborn rat following postnatal treatment with caffeine to mimic the therapeutic administration of caffeine to premature human infants. The effect of this postnatal exposure to caffeine on the gradual appearance of adenosine A1 receptors was analysed by determining immunohistochemically the distribution of the receptors. The main difference between control animals and animals exposed to caffeine was the transient increase (only at postnatal day 6) in the number of immunopositive neurons in two brainstem areas, the ventrolateral medulla and the rostral dorsolateral pons, in caffeine-treated rat pups, or more specifically, the parabrachial and Kolliker-Fuse nuclei, both of which are classically associated with respiratory control. With previous research highlighting the important role played by the rostral pons in respiratory modulation by the adenosine A1 receptor system, it is thus possible that postnatal exposure to caffeine modulates the ontogeny of the adenosine A1 receptor network. This could imply that the role of caffeine to decrease the incidence of neonatal respiratory disturbances may be due to the earlier than normal development of the adenosinergic system in the brain. PMID- 16702032 TI - Safety and efficacy of a new high power argon plasma coagulation system (hp-APC) in lesions of the upper gastrointestinal tract. AB - BACKGROUND: The aim of this study was to analyze safety and efficacy of a new high power argon plasma coagulation system in the upper gastrointestinal tract. METHODS: Data of 215 patients treated with a high power argon plasma coagulation system in the upper gastrointestinal tract 04/2003-01/2004, using a VIO APC device (VIO 300 D with APC 2; Erbe Elektromedizin, Tubingen, Germany; pulsed argon plasma coagulation, 20-120 W), were reviewed and analyzed. Indications were as follows: additive ablation therapy in curative treatment of early Barrett's cancer (122 patients); palliative treatment of oesophageal cancer (n=27); gastric adenoma/carcinoma (n=19); Zenker's diverticulum (n=8); and other. In 190/215 patients (149 males; mean age 67 years), the data were completely analyzable. Minor and major complications were evaluated. RESULTS: Minor complications (odynophagia, pain, fever) occurred in 24/277 sessions (8.7%); major complications (stenosis) in 3/277 sessions (1.1%) using at least 50 W. No perforation or bleeding occurred. The mean number of treatment sessions required was 1.46 (1-7); in the palliative treatment of oesophageal cancer, it was 2.5 (1 5). CONCLUSIONS: The high power argon plasma coagulation system was effective and safe in various gastrointestinal conditions. Due to it's high effectiveness and a low number of sessions required in tumour debulking, this high power argon plasma coagulation system might be used as an alternative to Nd:YAG laser. PMID- 16702033 TI - APACHE II score and primary liver cancer history had risk of hospital mortality in patients with pyogenic liver abscess. AB - BACKGROUND: The Acute Physiology and Chronic Health Evaluation II classification system has been extensively used for predicting the patient mortality in various diseases. However, its utilisation on the pyogenic liver abscess has not yet been well studied. AIMS: The purpose of this study was to validate this system on this high death rate disease. PATIENTS: A retrospective study was conducted to assess 314 patients with pyogenic liver abscesses admitted to tertiary medical centre in past 12 years. METHODS: The outcome measurement was the in-hospital mortality. A multiple logistic regression model was used to assess the association between mortality and Acute Physiology and Chronic Health Evaluation II score while controlling for the potential confounding factors. RESULTS: The overall in hospital mortality was 8.3%. The mean Acute Physiology and Chronic Health Evaluation II score of the expired patients was higher (P<0.0001). The mortality rate increased rapidly when Acute Physiology and Chronic Health Evaluation II score >or=15. After controlling for the potential confounding factors, patient with high admission Acute Physiology and Chronic Health Evaluation II score >or=15 had a higher chance of in-hospital mortality (P<0.01). In addition, the primary liver cancer history is also a risk factor (P=0.03). CONCLUSIONS: The Acute Physiology and Chronic Health Evaluation II score and the primary liver cancer history predict the in-hospital mortality of the pyogenic liver abscess patient. PMID- 16702034 TI - Subcutaneous rupture of hepatic hydatid cyst: CT findings. PMID- 16702035 TI - Prevention of food bezoar in the common bile duct by endoscopic stenting. PMID- 16702036 TI - Inflammation and bronchopulmonary dysplasia: a continuing story. AB - Increasing evidence indicates that bronchopulmonary dysplasia (BPD) results, at least in part, from an imbalance between pro-inflammatory and anti-inflammatory mechanisms, with a persistent imbalance that favours pro-inflammatory mechanisms. The inflammatory response is characterised by an accumulation of neutrophils and macrophages in the airways and pulmonary tissue of preterm infants and, moreover, by an arsenal of pro-inflammatory mediators which affect the alveolar capillary unit and tissue integrity. As well as pro-inflammatory cytokines and toxic oxygen radicals, various lipid mediators as well as potent proteases may be responsible for acute lung injury. During the last decade it has become evident that multiple pre- and postnatal events contribute to the development of BPD in preterm infants. Chorioamnionitis and cytokine exposure in utero, plus sequential lung injury caused by postnatal resuscitation, oxygen toxicity, volu-, barotrauma and infection all lead to a pulmonary inflammatory response which is most probably associated with aberrant wound healing and an inhibition of alveolarisation as well as vascular development in the immature lungs of very preterm infants, causing the 'new BPD'. PMID- 16702037 TI - Shear stress-induced pH increase in plasma is mediated by a decrease in P(CO(2)): the increase in pH enhances shear stress-induced P-selectin expression in platelets. AB - To investigate shear stress-induced platelet activation, the cone-plate viscometer or the Couette rotational viscometer has been widely used. In a previous report, it was shown that shearing platelet-rich plasma using a Couette rotational viscometer could lead to an increase in pH by CO(2) release. However, any clear mechanism has not been provided. In this study, we examined whether shearing cell free plasma only using a cone-plate viscometer can also induce pH increase and studied the underlying mechanism of shear-induced pH increase by directly measuring total CO(2) (T(CO(2))) and CO(2) tension (P(CO(2))). When human plasma was sheared using a cone-plate viscometer, the pH of the human plasma increased time- and shear rate-dependently. Although T(CO(2)) of human plasma was not affected, P(CO(2)) was decreased by shearing, indicating that the decreased P(CO(2)) is associated with a pH increase of plasma. In addition, the pH of bicarbonate-containing suspension buffer was also shown to be increased by shearing; suggesting that the platelet studies using suspension buffers containing bicarbonate could be affected similarly. The effects of pH changes on shear stress-induced platelet activation were also investigated in the same in vitro systems. While shear stress-induced platelet aggregation was not affected by the pH changes, P-selectin expression was significantly increased in accordance with the pH increase. In conclusion, shear stress using a cone-plate viscometer induces pH increase in plasma or bicarbonate-containing suspension buffer through a P(CO(2)) decrease and the pH changes alone can contribute to platelet activation by enhancing shear stress-induced P-selectin expression. PMID- 16702038 TI - Actin polymerisation regulates thrombin-evoked Ca(2+) signalling after activation of PAR-4 but not PAR-1 in human platelets. AB - The role of actin polymerisation in regulating thrombin-evoked Ca(2+) signalling was investigated in human platelets. We have previously reported that cytochalasin D (Cyt D) inhibits thapsigargin-evoked store-operated Ca(2+) entry (SOCE), which is believed to contribute a major component of thrombin-evoked Ca(2+) entry in platelets. In contrast, Cyt D increased thrombin-evoked Ca(2+) entry to 147.5 +/- 9.2% and Sr(2+) entry to 134.2 +/- 6.4% of control. Similar results were obtained with latrunculin A. This potentiation was not affected if protein kinase C was inhibited using Ro-31-8220, suggesting that it did not involve PKC-dependent non-capacitative Ca(2+) entry. Ca(2+) entry evoked by the PAR-4 agonist, AYPGKF, was increased to 133.7 +/- 12.8% of control by Cyt D, whereas Ca(2+) signalling evoked by the PAR-1 agonist, SFLLRN, was unaffected. The PAR-4 antagonist, tcY-NH(2), abolished the effect of Cyt D on thrombin-evoked Ca(2+) entry. Biotinylation of cell-surface proteins showed that PAR-4 was internalised after stimulation by thrombin. Cyt D reduced this internalisation. These data suggest that Cyt D prevents the internalisation of PAR-4, which may lead to prolonged signalling from this receptor. This may mask a direct effect of Cyt D on the activation of SOCE after the activation of PAR-4. PMID- 16702039 TI - Rosiglitazone-induced immune thrombocytopenia. AB - Rosiglitazone is one of the members in the thiazolidinedione (TZD) class of anti diabetic agents that have proven efficacy in the treatment of patients with type 2 diabetes. We studied serum from a patient who developed acute, severe thrombocytopenia after exposure to rosiglitazone maleate (Avandia) and proposed the mechanisms for rosiglitazone-induced thrombocytopenia. Tested by flow cytometry, the patient's serum was positive for rosiglitazone-induced antibody with the binding ratio of 5.93 (mean fluorescence intensity, MFI) in the presence of the patient's serum and rosiglitazone in a final concentration of 0.53 mmol/l. The antibody was found to bind both glycoprotein (GP) IIb-IIIa complex and GP Ib/IX complex by MAIPA assay using five different monoclonal antibodies (mAbs) against GP complexes Ib/IX, GPIIb/IIIa or GPIa/IIa. Immunoprecipitation studies showed that both GPIIb/IIIa and GP Ib/IX complex were precipitated by antibody in the presence, but not in the absence of rosiglitazone. These findings provide evidence that immune thrombocytopenia can be caused by sensitivity to the antidiabetic agent rosiglitazone maleate. This report documents the first case of rosiglitazone-induced immune thrombocytopenia. PMID- 16702040 TI - von Willebrand disease type 2B must be always considered in the differential diagnosis of genetic thrombocytopenias with giant platelets. AB - Type 2B von Willebrand's disease (VWD) is an inherited bleeding disorder characterized by spontaneous binding of large von Willebrand factor (VWF) multimers to platelets in vivo. This phenomenon induces the clearance of both large multimers and platelets, usually resulting in thrombocytopenia with slightly increased platelet size. We describe a newborn with a VWD type 2B due to the heterozygous missense mutation V1316M who presented the atypical feature of giant platelets in peripheral blood. Based on this observation and literature review, we suggest that the diagnosis of VWD 2B should be always considered in patients with chronic thrombocytopenia and giant platelets. PMID- 16702041 TI - Inhibition of platelet aggregation by grafting RGD and KGD sequences on the structural scaffold of small disulfide-rich proteins. AB - Disintegrins represent a group of disulfide-rich peptides ranging in size from 41 to over 80 residues and are antagonists of several integrin receptors. Disintegrins containing an RGD or KGD sequence are potent inhibitors of platelet aggregation as they block the binding of fibrinogen to alpha(IIb)beta(3) integrin. The high affinity binding to alpha(IIb)beta(3) in comparison to short linear peptides has been attributed to the localisation of the RGD or KGD sequence within a defined three-dimensional structure. Cystine knot microproteins are members of another family of small disulfide-rich peptides that consist of only 28-40 amino acid residues. They display numerous biological activities depending on the peptide sequence of loop regions that are fixed on a structural scaffold that is stabilised by three knot-forming disulfide bonds. In the present study we grafted RGD and KGD containing peptide sequences with seven and 11 amino acids, respectively, into two cystine knot microproteins, the trypsin inhibitor EETI-II and the melanocortin receptor binding domain of the human agouti-related protein AGRP, as well as into the small disintegrin obtustatin. The engineered proteins were much more potent to inhibit the fibrinogen binding, alpha(IIb)beta(3) activation and platelet aggregation when compared to the grafted peptides. Differences that were observed between the engineered proteins indicate the importance of the structural scaffold and the amino acids neighbouring the grafted peptide sequences. PMID- 16702042 TI - Functional interplay between platelet activation and endothelial dysfunction in patients with coronary heart disease. AB - Platelet-monocyte binding and surface P-selectin expression are sensitive markers of platelet activation. Endothelium-derived factors are known to inhibit platelet activation and may confer important anti-atherothrombotic effects. We assessed the relationship between platelet activation and endothelium-dependent vasomotion in patients with coronary heart disease (CHD). Twenty male patients with stable CHD were compared with 20 healthy men. Platelet-monocyte binding and platelet surface expression of P-selectin were assessed using two-colour flow cytometry on whole blood. Forearm blood flow was assessed in patients using venous occlusion plethysmography during intra-arterial infusions of substance P, acetylcholine and sodium nitroprusside. Platelet activation was higher in patients than healthy men (platelet-monocyte binding, 27 +/- 3 vs. 20 +/- 1%; P < 0.05). In patients with CHD, there was an inverse correlation between maximal substance P induced vasodilatation and both platelet-monocyte binding (P = 0.003) and P-selectin expression (P = 0.02). A similar correlation was observed between platelet monocyte binding and the vasomotor response to acetylcholine (P = 0.08) but not with sodium nitroprusside. In patients with stable coronary heart disease, there is a strong inverse relationship between markers of platelet activation and endothelium-dependent vasomotor function. This may explain the pathophysiological mechanism linking endothelial vasomotor dysfunction and the risk of acute atherothrombotic events. PMID- 16702043 TI - Aspirin resistance in coronary artery disease is correlated to elevated markers for oxidative stress but not to the expression of cyclooxygenase (COX) 1/2, a novel COX-1 polymorphism or the PlA(1/2) polymorphism. AB - Aspirin resistance (AR) is estimated to be present in 5-75% of patients and is related to increased cardiovascular mortality. However, the underlying mechanisms are mostly unknown. In the present study, AR was detected in 14 out of 55 patients (25%) with coronary artery disease. The presence of concomitant anti inflammatory drugs did not affect AR. Plasma levels of thromboxane B(2) as well as the markers for oxidative stress and known platelet activators 8-isoprostane and lipid peroxidation products were significantly higher in aspirin-resistant individuals (349.3 pg/ml, 53.9 pg/ml, and 538 micromol/l) compared to controls (113.7 pg/ml, 10.3 pg/ml, and 32.2 micromol/l; P < 0.05, respectively). Platelet cyclooxygenase-1 (COX-1) and COX-2 mRNA and protein expression were without significant differences between the two groups. DNA sequencing detected a novel platelet COX-1 single nucleotide polymorphism (SNP) resulting in amino acid exchange at position 8 (Arg8/Trp8). The wild-type as well as the heterozygous and homozygous SNP were present in both patient groups without significant differences. The aspirin binding (Arg120) and acetylation site (Ser529) were unaffected in the samples tested. Neither was AR related to the platelet integrin PlA(1)/A(2) polymorphism. In conclusion, AR appears to be unrelated to differences in platelet COX-1 and COX-2 expression or to a novel platelet COX-1 SNP and the PlA(1)/A(2) SNP. However, a correlation exists to elevated eicosanoids generated by oxidative stress indicating COX-1-independent pathways for the generation of platelet activating molecules represent a potential cause for AR. PMID- 16702044 TI - Evaluation of platelet turnover by flow cytometry. AB - The number of circulating newly produced platelets depends on the thrombopoietic capacity of bone marrow as well as platelet removal from the bloodstream. Flow cytometric analysis with thiazole orange (TO), a fluorescent dye that crosses platelet membranes and binds intracellular RNA, has been used to measure circulating reticulated platelets (RPs) with high RNA content as an index of platelet turnover. We first assessed the specificity of TO flow cytometry and then applied this method in the diagnosis of thrombocytopenia caused by impaired platelet production or increased destruction. We also explored the utility of TO flow cytometry to predict thrombocytopoiesis after chemotherapy-induced bone marrow aplasia. Venous blood, anticoagulated with K(2)EDTA, was incubated with 0.6 microg/ml TO plus an anti-GPIIIa monoclonal antibody. The mean percentage of RPs in control subjects (n = 23) was 6.13 +/- 3.09%. RPs were 10.41 +/- 9.02% in patients (n = 10) with hematological malignancies during aplasia induced by chemotherapy and a significant increase in RPs (35.45 +/- 6.11%) was seen in the recovery phase. In 10 patients with idiopathic thrombocytopenic purpura, the percentage of TO positive platelets was 67.81 +/- 18.79 (P < 0.001 vs. controls). In patients with thrombocytopenia associated with hepatic cirrhosis (n = 21; 21.04 +/- 16.21%, P < 0.001 vs. controls) or systemic lupus erythematosus (n = 6, 29.08 +/- 15.57%; P < 0.001 vs. controls) increases in TO-stained platelets were also observed. Measurement of TO positive platelets may be a reliable tool for the laboratory identification of platelet disorders, with a higher sensitivity than measurement of platelet volume. Measurement of RPs may also prove useful to recognize the underlying pathogenetic mechanisms in thrombocytopenia. PMID- 16702045 TI - Expression of functional recombinant mosquito salivary apyrase: a potential therapeutic platelet aggregation inhibitor. AB - Excessive platelet activation and accumulation can lead to vessel occlusion and thus present major therapeutic challenges in cardiovascular medicine. Apyrase, an ecto-enzyme with ADPase and ATPase activities, rapidly metabolizes ADP and ATP released from platelets and endothelial cells, thereby reducing platelet activation and recruitment. In the present study, we expressed a 68-kDa recombinant mosquito (Aedes aegypti) salivary apyrase using a baculovirus/insect cell expression system and purified it to homogeneity using anion-exchange chromatography on a large scale. A yield of 18 mg of purified recombinant apyrase was obtained from 1 litre of the medium. Kinetic analysis indicated that the recombinant apyrase had a K(m) of 12.5 microM for ADP and a K(m) of 15.0 microM for ATP. The recombinant apyrase inhibited ADP-, collagen- and thrombin-induced human platelet aggregation in a dose-dependent manner, indicating that the recombinant protein retained nucleotidase activity in a whole cell system, which suggests that it may serve as a therapeutic agent for inhibition of platelet mediated thrombosis. PMID- 16702046 TI - Differential platelet deposition onto collagen in cone-and-plate and parallel plate flow chambers. AB - To routinely test the formation of thrombi and the effect of drugs modifying it, proper test systems are needed. Their design should rely on the laws of rheology and the physiology of laminar flow. To best model physiological or pathological shear conditions, parallel/linear and rotational type flow chambers are developed. We have compared the initial phase of platelet thrombus formation in a parallel plate flow chamber (PPC) and a cone-and-plate chamber (CPC) under von Willebrand dependent shear conditions. Blood was allowed to flow through human collagen type III surfaces at a shear rate of 1000 s(-1) for 150 s. Thrombus deposition was characterized by surface coverage, average area and height of thrombi. VWF distribution within thrombi was analyzed with confocal laser scanning microscopy. Reduced surface-specific platelet adhesion and aggregation (surface coverage and average thrombus size) were observed in CPC along with a significant increase in single platelet disappearance from the circulating blood. Our data suggest that the higher rate of platelet consumption in this device, as opposed to PPC, is limiting the adhesion to the surface. Consequently, surface specific processes and aggregation in the flowing blood are both assessed using CPC, while comprehensive evaluation of surface-specific processes is best achieved with PPC. Therefore, the choice of chamber type as a diagnostic tool is purpose-dependent. PMID- 16702047 TI - Why human platelets fail to kill bacteria. AB - Recent investigations have suggested that bacteria are taken up by platelets in the same manner as by polymorphonuclear (PMN) leukocytes and monocytes. The phagocytic vacuole containing the organism is completely separated from the cell exterior and the cytoplasm in order to form a killing chamber. Yet, earlier studies demonstrated that platelets do not kill bacteria. The present work has provided a basis for the lack of platelet bactericidal activity. Platelets incubated with Staphylococcus aureus 502A or RN 450 for intervals up to 2 h can take up organisms into sequestration vacuoles that resemble the phagosomes formed by PMN and monocytes. However, staining with tannic acid which forms an electron dense stain with osmic acid reveals that the phagosomes in PMN and monocytes are completely separated from the cell exterior and the cytoplasm. As a result, they are true killing chambers. The engulfment vacuoles containing bacteria in platelets on the other hand are almost never sealed from the cell exterior. Osmium black reaction product stains the inside of the engulfment vacuole and channels of the open canalicular system connecting it to the cell exterior. Thus platelets do not form the killing chamber phagosomes observed in leukocytes and, as a result, cannot kill bacteria. PMID- 16702048 TI - The free-radical scavenger, edaravone, augments NO release from vascular cells and platelets after laser-induced, acute endothelial injury in vivo. AB - In vitro and in vivo experimental models have demonstrated that vascular endothelial function is significantly impaired as a result of oxidative stress, mediated by the generation of oxygen-derived free radicals in response to chronic or acute inflammation. In particular, super-oxide () at specific concentrations leads to the impairment of nitric oxide (NO) bioactivity, and it is known that NO plays a fundamental role in the maintenance of vascular homeostasis. The relationship between reactive oxygen species (ROS) and NO release in thrombosis related endothelial damage in the peripheral microvasculature remains unclear, however. The purpose of the present study was to investigate the effect of the free-radical scavenger, edaravone, on NO synthesis and thrombotic potential in arterioles after exposure to laser irradiation. Highly sensitive electrochemical NO microsensors were positioned in femoral arterioles of mice, and the kinetics of NO release were recorded in response to standardized laser irradiation in vivo. In addition, images of NO release from damaged vascular cells were investigated in a similar rat model using the NO-sensitive dye 4,5 diaminofluorescein diacetate (DAF-2DA). Thrombogenesis was assessed in carotid arterioles by continuous video microscopy using image analysis software. Laser irradiation led to NO release from perturbed endothelial cells and from platelet rich thrombi. Edaravone had no significant effect on NO release in non-laser treated, intact endothelium compared with placebo. In contrast, edaravone demonstrated a dose-dependent effect on NO release and thrombogenicity. At a concentration of 10.5 mg/kg per h, edaravone promoted a 5-fold increase in NO and a reduction in platelet-rich thrombus volume to 58% of the placebo values. Our data provide direct evidence to confirm that acute endothelial damage in peripheral microvessels initially induces NO release and that the free-radical scavenger, edaravone, augments NO synthesis leading to suppression of platelet thrombus formation. PMID- 16702049 TI - Fiberoptic bronchoscopy induces platelet activation. PMID- 16702050 TI - Drug-induced psoriasis: an evidence-based overview and the introduction of psoriatic drug eruption probability score. AB - Psoriasis is a common skin disorder that needs a long-term management, not only because, of its prevalence but also because of the profound impact it can have on patients quality of life. Drugs may result in exacerbation of a preexisting psoriasis, in induction of psoriatic lesions on clinically uninvolved skin in patients with psoriasis, or in precipitation of the disease in persons without family history of psoriasis or in predisposed individuals. The knowledge of the drugs that may induce, trigger, or exacerbate the disease is of primary importance in clinical practice. By reviewing the literature, there are many reports on drug-induced psoriasis, but the data are not univocal. We propose, when possible, the use of a probability score from the authors to obtain a better classification and further understanding of drug-induced psoriasis. PMID- 16702051 TI - Xenobiotics in vitro: the influence of L-cystine, pantothenat, and miliacin on metabolic and proliferative capacity of keratinocytes. AB - To investigate the effect of cell growth-stimulating agents on human epidermal keratinocytes, we exposed monolayers of normal human keratinocytes derived from foreskin to different concentrations of the amino acid L-cystine, the member of the vitamin B family D-pantothenat, the phytosterol miliacin, and a combination thereof in keratinocyte growth medium. As a test system for the metabolic capacity, we used the activity of mitochondrial deyhdrogenases as measured by XTT, and for the cell proliferation, we determined the BrdU-uptake. The additives, active ingredients of the hair growth drug PRIORIN, were added in the presence of fully supplemented keratinocyte growth medium or a deficient medium without L-cystine, L-methionine, L-histidin, D-pantothenat, epidermal growth factor, and bovine pituary gland extract. Deficient medium itself reduced the metabolic capacity of keratinocytes to 35% compared with keratinocytes in fully supplemented growth medium. In deficient medium cell, proliferation was not measurable. Increasing doses of L-cystine restored the reduced metabolic capacity from 46% (0.009 mg/L) to 54% (0.09 mg/L) and 92% (0.45 mg/L) in deficient medium. Addition of D-pantothenat (0.43 mg/L) enhanced the metabolic capacity to 150% only in fully supplemented growth medium, compared with untreated controls with growth medium. Miliacin (6 mg/mL) increased not only the metabolic capacity (162%) but also stimulated cell proliferation (215%) as measured by BrdU-uptake in growth medium. The combination of all three additives increased the metabolic capacity (245%) synergistically in growth medium. We were able to show effects of D-panthenol, L-lysine, and miliacin on proliferation and metabolic capacity of keratinocyte monocell culture, which was further increased by combination of the three substances. These basic results suggest a beneficial effect on keratinocyte growth and stimulation by products combining these substances (e.g., Priorin). Furthermore, this work emphasizes the suitability of keratinocyte monolayers for pharmacological testings. PMID- 16702052 TI - Textiles and human skin, microclimate, cutaneous reactions: an overview. AB - This article overviews research in the interdisciplinary area of textile/skin interaction and related cutaneous intolerance. Microclimate in the skin/clothing system and especially the skin responses relates to the moisture and heat transfer within this system and plays a critical role in skin irritation from textiles. A discussion is then given on skin irritation reactions to textiles, including intolerance caused by chemicals (dyes and finishes) and physical contact/friction. Finally, two skin injuries, blisters and pressure ulcers, which are caused by physical contact, pressure, and friction, are documented. Despite the prevalent problems caused by ill textile/skin interactions, minimal efforts have been devoted to this field. In addition, the in vivo experimental studies infrequently lead to a solid conclusion. The cause may lie in the dramatic variation of skin conditions among individuals as well as among different anatomic sites of the same person. Another reason might be the lack of communications between researchers in the areas of textiles and dermatology. PMID- 16702053 TI - Extent of corneal injury as a biomarker for hazard assessment and the development of alternative models to the Draize rabbit eye test. AB - We have characterized 22 ocular irritants differing in type (surfactants, acid, alkali, bleaches, alcohol, aldehyde, acetone) and severity (slight to severe) by using the low-volume rabbit eye test. Ocular irritation was evaluated by 1) light microscopy to assess pathological changes, 2) in vivo confocal microscopy (CM) to quantify 4-dimensionally (x, y, z, and t) initial corneal injury and later responses in the same eye, and 3) laser scanning CM to quantify initial cell death. These studies revealed that regardless of the processes leading to injury, slight irritants injure the corneal epithelium, mild irritants injure the corneal epithelium and the superficial stroma, and moderate/severe irritants injure the epithelium, deep stroma, and at times the corneal endothelium. Furthermore, extent of initial corneal injury was shown to predict subsequent responses and final outcomes. These findings suggest that extent of corneal injury may be used as a basis for the development of alternative ocular irritation tests. To test the validity of this approach, we have used an ex vivo, rabbit cornea culture model to measure extent of corneal injury following exposure to ocular irritants. Data indicate that the extent of ex vivo corneal injury significantly correlate with the extent of initial injury measured previously in live animals. Overall, these findings indicate that extent of initial corneal injury can be used as a new "gold standard" for the continued refinement and ultimate replacement of the Draize rabbit eye Ocular Irritation Test. PMID- 16702054 TI - Sulfuric acid burns (corrosion and acute irritation): evidence-based overview to management. AB - Concentrated sulfuric acid causes severe skin injury. To prevent skin destruction, efficient early treatment is of utmost importance. However, regimens suggested in the literature are not always supported by experimental data. Further studies are needed. To improve early management of sulfuric acid burns, future experiments need careful extrapolation between animal skin and human skin. The benefit of water, neutralizer, or alternative agents has to be established by precisely defining acid concentration and time of exposure. PMID- 16702056 TI - Synthetic dairy wastewater treatment using a new horizontal-flow biofilm reactor. AB - The aim of this study was to develop a simple biological system that would be suitable for the treatment of dairy parlour wash waters. A novel horizontal-flow biofilm system was designed, constructed and tested in the laboratory for organic carbon and nitrogen removal from a synthetic dairy wastewater with average filtered chemical oxygen demand (CODf) of 2947.7 mg/L and total nitrogen (TN) of 295.5 mg/L. The novel biofilm system consisted of two reactor units one on top of the other. Each reactor unit comprised a stack of horizontal plastic sheets placed above one another that were supported and separated by vertical plastic cone frustums, which were formed in the sheets during manufacture. The wastewater was pumped onto the top sheet of the top reactor unit at hourly intervals for a 5 minute period and flowed over and back along alternate sheets down through the 2 units. The top unit had 8 plastic sheets and 25 mm high frustums to cater for heavy biofilm growth, and the bottom unit had 17 sheets and 11 mm high frustums for thinner biofilm growth. The organic loading of 57 g CODf/m2.d-based on the plan area of the system-was applied to the top reactor unit. When the reactor process reached pseudo steady-state, about 96% CODf, 71% TN and 100% ammonium nitrogen were removed. Low solids production occurred. The system was simple and easy to construct and operate. PMID- 16702057 TI - Bioaugmentation of activated sludge with two Pseudomonas putida strains for the degradation of 4-chlorophenol. AB - The augmentation of activated sludge with two strains of Pseudomonas putida, CP1 and A(a) was investigated. Both strains of bacteria degraded 1.56 mM 4 chlorophenol. P. putida CP1 degraded the chemical using a modified ortho-cleavage pathway while P. putida A(a) used the meta-cleavage pathway. When activated sludge incapable of 4-chlorophenol degradation was augmented with either strain, substrate degradation occurred and followed the same biochemical pathways as when the bacteria were grown in pure culture. Insertion, in tandem, of the genes for gentamycin resistance and green fluorescent protein into the chromosomes of the two strains, enabled the survival and spatial location of the bacteria in the mixed microbial population to be monitored. Labelling the bacteria did not alter their degradative capabilities. P. putida CP1::Tn7-gfp survived in higher numbers than P. putida A(a)::Tn7-gfp following addition to the activated sludge. This was attributed to the ability of this strain to flocculate and become integrated in the activated sludge floc. Addition of P. putida CP1::Tn7-gfp or A(a)::Tn7-gfp to activated sludge resulted in smaller decreases in total cell numbers indicating a protective effect of the introduced P. putida strains on the overall microbial population from the harmful effects of 4-chlorophenol. The non-flocculant strain A(a) did not survive as well as CP1 in the activated sludge system and required a higher inoculum size to effect substrate degradation. PMID- 16702058 TI - Denitrification of a nitrate-rich synthetic wastewater using various wood-based media materials. AB - This laboratory study examined the use of various wood materials as a carbon source in horizontal flow filters to denitrify nitrate-nitrogen (NO3-N) from a synthetic wastewater. The filter materials were: sawdust (Pinus radiata), sawdust and soil, sawdust and sand, and medium-chip wood chippings and sand. Two influent concentrations of NO3-N, 200 mg L(-1) and 60 mg L(-1), loaded at 2.9 to 19.4 mg NO3-N kg(-1) mixture, were used. The horizontal flow filter with a wood chippings/sand mixture and an influent NO3-N concentration of 60 mg L(-1), which operated over a study duration of 166 days, performed best, yielding a 97% reduction in NO3-N at steady-state conditions. PMID- 16702059 TI - Comparison of selected physicochemical characteristics of commercial phytases relevant to their application in phosphate pollution abatement. AB - The enzyme phytase catalyses the dephosphorylation of phytic acid and it's salts, phytates. Supplementation of monogastric animal feed with microbial-derived phytase increases the bioavailability of phytic acid bound phosphate. This facilitates a reduction in the addition of inorganic phosphate to the feed and reduces phosphorus excretion. To achieve maximum efficacy in terms of phosphate pollution abatement, supplemental phytases added to animal feed must survive thermal processing of the feed, resist inactivation by the proteolytic enzymes encountered in the animal's digestive tract and display high activity at physiological temperature and pH. A series of in vitro experiments were carried out to determine the relative suitability of four major commercial phytase products for use in animal feed. The enzymes assessed lost between 14% and 72% of their original activities after heating to 80 degrees C for 5 minutes. After exposure to simulated upper digestive tract conditions, the phytases assessed retained between 0 and 28% of their original activities. The commercial phytases displayed between 98% and 67% of their maximum activities at 39 degrees C and all phytases assessed had an optimum pH between pH 4 and pH 5. None of the phytases assessed satisfied all of the criteria of an ideal phytase for use in animal feed. PMID- 16702060 TI - Effects of the smoking ban on benzene and 1,3-butadiene levels in pubs in Dublin. AB - According to World Health Organisation figures, 30% of all cancer deaths, 20% of all coronary heart diseases and strokes and 80% of all chronic obstructive pulmonary disease are caused by cigarette smoking. In accordance with the recommendations of the Tobacco Free Policy Review Group Report the Irish government has introduced a smoking ban in all workplaces with the exception of prisons and psychiatric hospitals. This study measured the levels of benzene and 1,3-butadiene in air, two known carcinogens and environmental tobacco smoke (ETS) markers, in pubs both before and after the smoking ban was introduced. The results of the study have quantified the significant gross differences in pre and post ban exposure levels. The International Commission on Radiological Protection (ICRP), Human Respiratory Tract model for Radiological Protection has then been adopted to assess the amounts of these pollutants typically absorbed in the nose, throat and lungs of the workers and patrons of pubs in Ireland before and after the smoking ban. This has revealed a reduction in the average dose of benzene and 1,3-butadiene of 91% and 95% respectively for a typical three hour exposure in Irish pub. PMID- 16702061 TI - The performance of fibrous peat biofilters in treating domestic strength wastewater. AB - Peat is an abundant resource in Ireland and has the capacity to be used in low cost, low-maintenance wastewater treatment systems for single houses. In this study four fibrous peat columns, of varying depths were constructed and tested in the laboratory for their capacity to remove contaminants from domestic-strength synthetic wastewater. The four filters had peat depths of 0.3 m, 0.6 m, 0.9 m and 1.2 m. During the 360 day study the filters were intermittently loaded with domestic strength synthetic wastewater at a hydraulic loading rate of 180 l/m2cd. Hydrographs and residence times for each filter were examined as was their ability to remove impurities from the wastewater. Removal of 5-day biochemical oxygen demand (BOD5) and total chemical oxygen demand (CODt) were > or =96% and 84%, respectively, in all filters with almost complete nitrification (> or =99%) being recorded for each filter. The removal of total suspended solids (TSS) was excellent at > or =94% and no clogging was recorded on any filter during the study. For the 0.6 m, 0.9 m and 1.2 m deep filters, total viable counts (TVC) were less than EU surface water directive limits for Class A2 potable water sources. The systems were cheap to construct and very easy to maintain. PMID- 16702062 TI - Modelling the fate and transport of nickel in the mersey estuary. AB - Modelling heavy metals in estuarine environments is extremely complex for various reasons; one of the primary complicating factors is that metals exist in two phases, dissolved and particulate bound. Dynamic changes in water chemistry, and in particular salinity, affect the partitioning of metals between the two phases and hence make it difficult to determine the relative fractions of each phase. A relatively simple approach was developed to relate variations in partition coefficient for Ni to salinity fluctuations in the Mersey Estuary. The functional relationship developed between partition coefficient and salinity departs from the traditional exponential type curve, providing a more realistic relationship.A numerical model was then developed for predicting the transport and distribution of Ni about the Mersey Estuary. The model couples transport of metals throughout the water along with incorporating the chemical processes controlling how nickel is fractioned between dissolved and particulate phases through the newly developed partition coefficient relationship. Model predictions of dissolved Ni along the longitudinal axis of the estuary were compared with measurements of Ni for two events; very good correlation was obtained between the model results and the data. PMID- 16702064 TI - A technical, economic and environmental comparison of composting and anaerobic digestion of biodegradable municipal waste. AB - Ireland produces two million tonnes per annum of biodegradable municipal solid waste. The implementation of the Landfill Directive (1999/31/EC) will lead to the construction of centralised biological facilities; these facilities may be either composting facilities or anaerobic digestion facilities. A technical, economic and environmental analysis of composting and anaerobic digestion is undertaken in this paper. The results of the analysis suggest that composting is economically preferable to anaerobic digestion at scales at or below 50 kt/a of biowaste treated. However when CH4-enriched biogas is produced for use as a transport fuel and excise duty is reduced, as allowed by the Biofuels Directive (2003/30/EC), then the economics of anaerobic digestion improve greatly. If 100% of excise duty is removed then anaerobic digestion is economically preferable to composting above 20 kt/a of biowaste treated. From an environment perspective anaerobic digestion saves more greenhouse gas due to displacement of fossil fuel powered energy. Anaerobic digestion with CH4-enriched biogas has the potential to save 1,451 kgCO2/t of biowaste treated as opposed to composting, which has the potential to save 1,190 kgCO2/t. PMID- 16702063 TI - Trophic status of experimental cutaway peatland lakes in ireland and implications for future lake creation. AB - Of the 80,000 hectares of Bord na Mona owned peatland coming out of industrial production in Ireland approximately the next 25 years, over 20,000 hectares has been designated for shallow lake creation. Four experimental lakes created by flooding areas of redundant cutaway peatland in Co. Offaly were monitored over a 3-year period in order to obtain baseline information on their water quality and trophic status. Results indicate that water chemistry in the constructed lakes was predominantly influenced by the depth and type of the residual peat layers at the sites, the degree of exposure of underlying inorganic subsoils and the type of hydrological regime. Nutrient status was strongly governed by catchment land uses. Lack of recolonising vegetation at recently abandoned cutaway peatland sites made some new lakes particularly vulnerable to nutrient runoff and algal bloom development. Biologically, the embryonic lakes were characterised by rudimentary food chains, in which higher trophic levels were absent and where the microbiota played an elevated role. PMID- 16702065 TI - New low-temperature applications of anaerobic wastewater treatment. AB - Low-temperature or psychrophilic (<20 degrees C) anaerobic biological treatment of simple industrial wastewaters has recently been proven feasible as an alternative to more expensive mesophilic (ca. 37 degrees C) technology. We implemented novel expanded granular sludge bed (EGSB)-based bioreactor designs for 27 psychrophilic anaerobic digestion (PAD) trials for the treatment of a broad range of simple and complex synthetic wastewaters representing dairy, food processing and pharmaceutical sector effluents. A variety of operating parameters, such as hydraulic retention time, organic and volumetric loading rates and upflow velocity, were tested. Chemical oxygen demand (COD) removal efficiencies were recorded, which were comparable to previous mesophilic trials. Specific methanogenic activity, toxicity and biodegradability batch assays were employed to monitor the metabolic capabilities of microbial consortia in anaerobic reactors. The prevalence of psychrotolerant communities was observed and psychrophilic populations were detected in two of the reactors. The potential of PAD with respect to global sustainable development is discussed. PMID- 16702066 TI - Accessing the black box of microbial diversity and ecophysiology: recent advances through polyphasic experiments. AB - The microbial ecology of a range of anaerobic biological assemblages (granular sludge) from full- and laboratory-scale wastewater treatment bioreactors, and of crop-growing and peat soils, was determined using a variety of 16S rRNA gene based techniques, including clone library, terminal restriction fragment length polymorphism (TRFLP) and denaturing gradient gel electrophoresis (DGGE) analyses. Fluorescent in situ hybridization (FISH) using 16S rRNA gene-targeted probes was employed to complete a "full-cycle rRNA approach" with selected biomass. Genetic fingerprinting (TRFLP and DGGE) was effectively used to elucidate community structure-crop relationships, and to detect and monitor trends in bioreactor sludge and specific enrichment cultures of peat soil. Greater diversity was resolved within bacterial than within archaeal communities, and unexpected reservoirs of uncultured Crenarchaeota were detected in sludge granules. Advanced radiotracer incubations and micro-beta imaging were employed in conjunction with FISH to elucidate the eco-functionalism of these organisms. Crenarchaeota clusters were identified in close associated with methanogenic Archaea and both were localised with acetate uptake in biofilm structure. PMID- 16702067 TI - Comparing primary energy attributed to renewable energy with primary energy equivalent to determine carbon abatement in a national context. AB - The current conventional approach to determining the primary energy associated with non-combustible renewable energy (RE) sources such as wind energy and hydro power is to equate the electricity generated from these sources with the primary energy supply. This paper compares this with an approach that was formerly used by the IEA, in which the primary energy equivalent attributed to renewable energy was equated with the fossil fuel energy it displaces. Difficulties with implementing this approach in a meaningful way for international comparisons lead to most international organisations abandoning the primary energy equivalent methodology. It has recently re-emerged in prominence however, as efforts grow to develop baseline procedures for quantifying the greenhouse gas (GHG) emissions avoided by renewable energy within the context of the Kyoto Protocol credit trading mechanisms. This paper discusses the primary energy equivalent approach and in particular the distinctions between displacing fossil fuel energy in existing plant or in new plant. The approach is then extended provide insight into future primary energy displacement by renewable energy and to quantify the amount of CO2 emissions avoided by renewable energy. The usefulness of this approach in quantifying the benefits of renewable energy is also discussed in an energy policy context, with regard to increasing security of energy supply as well as reducing energy-related GHG (and other) emissions. The approach is applied in a national context and Ireland is case study country selected for this research. The choice of Ireland is interesting in two respects. The first relates to the high proportion of electricity only fossil fuel plants in Ireland resulting in a significant variation between primary energy and primary energy equivalent. The second concerns Ireland's poor performance to date in limiting GHG emissions in line with its Kyoto target and points to the need for techniques to quantify the potential contribution of renewable energy in achieving the target set. PMID- 16702068 TI - A two-species biomarker model for the assessment of sediment toxicity in the marine and estuarine environment using the comet assay. AB - Sediments frequently cause damage to biota due to the accumulation of toxic compounds and the bioavailability of sediment-associated contaminants. Damage can be assessed using biomarkers, such as the degree of genotoxic impact following in vivo exposure to contaminants. Genotoxic damage, expressed as single-strand DNA breaks, was measured in cells isolated from haemolymph/blood, gill and digestive gland/liver from the clam Tapes semidecussatus and turbot Scophthalmus maximus, using the single cell gel electrophoresis (Comet Assay). Both animals were exposed for three weeks to sediment samples collected from a polluted site and a 'clean' reference site. The level of DNA damage was assessed using an image analysis package and expressed as % tail DNA. Throughout the study, significant differences in DNA damage were recorded for each tissue type, in both species, between animals exposed to the two sediment samples. However, turbot appeared to be a more sensitive indicator species, because, due to lower background levels, they were able to detect a significant difference between reference site and background values. This suggests that turbot, rather than clams, are more suitable as a sentinel species for the assessment of genotoxic impact of low level contamination in aquatic sediments and highlights the need for a two- or multi-species approach. PMID- 16702071 TI - A developmental model of infantile nystagmus. AB - The possibility that infantile nystagmus (IN) may reflect a failure in early sensorimotor integration has been proposed for more than a century, but is only recently being borne out in animal studies. The underlying neural and genetic substrate for this plasticity is complex. We propose that, in most cases, IN develops as a developmental response to reduced contrast sensitivity to high spatial frequencies in an early "critical period," however caused, whether by structural malformations (e.g. foveal hypoplasia) or poor optics (e.g. cataract). As shown by psychophysics, contrast sensitivity to low spatial frequencies is enhanced by motion of the image across the retina. Based on our previous theoretical study (Harris & Berry, Nonlinear Dynamics, 2006), we argue that the best compromise between moving the image and maintaining the image near the fovea (or its remnant) is to oscillate the eyes with jerk nystagmus with increasing velocity waveforms, as seen empirically. The generation of jerk waveforms relies heavily on the saccadic system, which is immature in infancy. Pendular waveforms may therefore provide an alternative to jerk waveforms, and may explain why they are seen more often in young infants. We discuss the implications of this developmental model for the need to synchronize sensory and motor developments in normal development. Failure of this synchronization may also explain some idiopathic cases. PMID- 16702072 TI - Biologically relevant models of infantile nystagmus syndrome: the requirement for behavioral ocular motor system models. AB - Infantile nystagmus syndrome (INS) is a combination of several types of nystagmus, each representing dysfunction in one subsystem of the ocular motor system (OMS) and having characteristic waveforms. Eye-movement recordings are the only certain way to identify IN and differentiate it from other types. The waveform classification scheme in use for 30 years is both accurate, inclusive, and suggests the underlying subsystem instabilities. In different individuals, they may appear at birth (hard wired) or in early infancy (developmental). The primary subsystem instability in IN is hypothesized to lie in the normally underdamped smooth pursuit system; vestibular dysfunction (imbalance) may also be present. Less often, the nucleus of the optic tract may be involved. Ocular motility studies over the past 40 years have demonstrated that saccades and gaze holding are normal in the INS and saccades contained within IN waveforms are always corrective; i.e., they cannot be the initiating movement responsible for IN. Because there are an infinite number of solutions to simulating specific waveforms, models that merely generate waveforms that resemble IN in isolation are of little use, either clinically or to increase our understanding of the underlying mechanisms of IN. A biologically relevant model of the INS should be part of, and operate within, a complete OMS model, capable of reproducing the normal ocular motor behavior of these individuals while still oscillating; i.e., the model, like the patient, must not have oscillopsia and be able to respond correctly to various target inputs. PMID- 16702073 TI - Modelling Early Onset Nystagmus (EON). AB - Six presentations at the Nystagmus Research Workshop (Abingdon 2005) were delivered in the session on 'Modelling'. The following is a brief summary of these presentations, indicating (i) the wide range of approaches taken, (ii) the rationale for each approach, and (iii) possible experiments to help resolve apparent conflicts between differing theoretical models of Early Onset Nystagmus. PMID- 16702074 TI - What can acquired nystagmus tell us about congenital forms of nystagmus? AB - For several forms of acquired nystagmus, animal models exist, mathematical hypotheses have been proposed, and treatments are available. What insights could acquired nystagmus provide for congenital forms of nystagmus? Acquired periodic alternating nystagmus (PAN) is caused by instability of the velocity storage mechanism for vestibular eye movements; an adaptive mechanism produces the oscillations that have a period of about 4 minutes. Surprisingly, the ability of individuals with congenital forms of nystagmus to adapt their eye movements to new visual demands has received little study. Acquired pendular nystagmus (APN) may arise from instability in the neural integrator for eye movements; identification of the neurotransmitters contributing to normal gaze holding made it possible to identify candidate drugs for treatment of APN. Similar knowledge of the biology underlying of congenital forms of nystagmus might similarly suggest effective drugs. Downbeat nystagmus (DBN) is caused by cerebellar disease, which includes structural lesions affecting the flocculus and paraflocculus, and calcium channelopathies, such as episodic ataxia type 2 (EA2), for which a mouse model and effective treatment is available. Since some congenital forms of nystagmus are genetic in origin, then the possibility arises that they may be caused by a channelopathy, a hypothesis that suggests novel drugs for evaluation in randomized controlled trials. PMID- 16702075 TI - The molecular genetics of congenital idiopathic nystagmus. AB - Congenital Idiopathic Nystagmus (CIN) is genetically heterogeneous. Autosomal dominant, autosomal recessive and X-linked patterns of inheritance have been reported. Linkage analysis has suggested the existence of at least three distinct loci for both autosomal dominant and x-linked forms, although as yet no disease genes have been identified. The pathophysiological mechanisms underlying nystagmus are poorly understood and it is likely that insights may arise from finding and characterizing disease genes. If linkage experiments are used to find "nystagmus genes," their power will depend heavily on accurate phenotyping to avoid misdiagnosis due to masquerading conditions and phenotypic variations within pedigrees. PMID- 16702077 TI - Predicting visual acuity in early onset nystagmus. AB - In preverbal children, it is difficult to assess visual acuity accurately. If nystagmus waveforms can be used to predict visual acuity in these young patients, then they would be a useful additional clinical tool. Existing acuity predictors were assessed in 10 adults with idiopathic early onset nystagmus. The most reliable predictor in this group of patients was mean velocity of the waveform. The mean velocity of the waveform may be used in preverbal children to give an estimate of best potential visual acuity in adulthood, and estimated best potential visual acuity at the current age. PMID- 16702076 TI - Visual and perceptual consequences of congenital nystagmus. AB - One way to assess the influence of retinal image motion on visual functioning in congenital nystagmus (CN) is to examine the effects of comparable image motion in observers with normal vision. A second approach is to evaluate visual functioning in subjects with CN when the retinal image motion is reduced. Using these approaches, we determined that spatial contrast sensitivity and visual acuity are not limited by the parameters of retinal image motion in some subjects with CN, but rather by a form of amblyopia. Clinical evidence from patients with bilateral refractive amblyopia suggests that a gradual improvement of visual acuity may be possible in persons with CN, if the optimal refractive correction is worn and parameters of the retinal image motion undergo long-term amelioration. PMID- 16702078 TI - Clinical and electrophysiological effects of extraocular muscle surgery on patients with Infantile Nystagmus Syndrome (INS). AB - The purpose of this report is to summarize clinical and electrophysiological effects of extraocular muscle surgery in patients with INS. Our hypothesis is that surgery on the extraocular muscles of patients with INS changes their nystagmus resulting in improved vision and visual functions. All patients had all four virgin horizontal recti operated on, either for strabismus alone, nystagmus alone, for a head posture due to an eccentric null zone alone or for a head posture due to an eccentric null zone plus strabismus. All patients have been followed for at least 12 months. Subjective outcome measures include the pre- and post-operative binocular best optically corrected null zone acuity (NZA) in 75 patients and gaze dependent acuity (GDA) in 12 patients. Objective outcome measure included null zone width (NZW) in 75 patients. The results are summarized as follows: NZA increased .1 LogMar or greater in 75% with those patients or=90% of patients and in their principal therapeutic targets in a cohort receiving their first focal spasticity treatment with botulinum toxin A and additional therapy. A strict strategy for patient selection and comprehensive management was followed. PMID- 16702084 TI - A comprehensive pain management programme comprising educational, cognitive and behavioural interventions for neuropathic pain following spinal cord injury. AB - OBJECTIVE: To assess whether a comprehensive multidisciplinary pain management programme could contribute to improvement regarding sleep quality, mood, life satisfaction, health-related quality of life, sense of coherence and pain for patients with a spinal cord injury and neuropathic pain. DESIGN: A prospective intervention study. PATIENTS: Twenty-seven patients with spinal cord injury and neuropathic pain participated in a pain management programme in parallel with 11 patients in a control group. METHODS: A comprehensive pain management programme comprising educational, cognitive, and behavioural interventions was created for patients with spinal cord injury and neuropathic pain. The pain management programme consisted of 20 sessions over a 10-week period and included educational sessions, behavioural therapy, relaxation, stretching, light exercise and body awareness training. All patients were followed-up 3, 6 and 12 months after completion of the programme. RESULTS: At the 12-month follow-up, levels of anxiety and depression in the treatment group decreased compared with baseline values, and a tendency towards better quality of sleep was seen. In comparison with the control group, patients in the treatment group improved regarding sense of coherence and depression. CONCLUSION: This study implies that a multidimensional pain management programme can be a valuable complement in the treatment of spinal cord injured patients with neuropathic pain. PMID- 16702085 TI - Health-related quality of life in myotonic dystrophy type 1 and its relationship with cognitive and emotional functioning. AB - OBJECTIVE: To evaluate the health-related quality of life in myotonic dystrophy type 1 and its relationships with clinical, genetic, neuropsychological and emotional factors. DESIGN: Case-control study of a continuous series of patients with myotonic dystrophy type 1. PATIENTS AND METHODS: Twenty patients, and 20 age , sex- and education-matched healthy controls underwent the MOS 36-Item Short Form Health Survey (SF-36), an extensive neuropsychological battery and emotional functioning tests. RESULTS: Patients' SF-36 mean scores were lower than those of controls in all dimensions. The neuropsychological study showed a significant impairment in visuospatial and verbal abstract reasoning (p=0.001), visuospatial memory (p=0.002) and attentive functions (p=0.03) in patients with myotonic dystrophy type 1. The emotional assessment showed significantly high scores in anxiety (p=0.002) and depression (p=0.001), which occurred in approximately 50% of patients. Both physical and mental SF-36 areas were inversely correlated with age, duration and grade of disease, depression and anxiety and positively correlated with attentive control. SF-36 areas were not correlated with cytosine thymine guanidine expansion. CONCLUSION: Health-related quality of life is severely impaired in myotonic dystrophy type 1 and it is negatively influenced by severity and duration of disease as well as by specific cognitive deficits and changes in emotional functioning. Therapeutic intervention in this field could contribute to ameliorate health-related quality of life in myotonic dystrophy type 1. PMID- 16702086 TI - How to identify potential fallers in a stroke unit: validity indexes of 4 test methods. AB - OBJECTIVE: The aim of this study was to describe general characteristics of patients with stroke who have a tendency to fall and to determine whether certain test instruments can identify fallers. METHODS: Patients treated in a stroke unit during a 12-month period were included. At inclusion assessments were made with Berg Balance Scale Berg Balance Scale, Stops Walking When Talking, Timed Up & Go (TUG) and diffTUG. At follow-up 6 or 12 months later, patients who had fallen were identified. RESULTS: During the time from discharge to follow-up on 159 patients, 68 patients fell and 91 did not. Fallers fell more often during their initial hospital stay, used sedatives more often and were more visually impaired, compared with non-fallers. The Berg Balance Scale, Stops Walking When Talking and TUG results differed between fallers and non-fallers. The combined results of Berg Balance Scale and Stops Walking When Talking increased the possibility of identifying fallers. CONCLUSION: Berg Balance Scale, Stops Walking When Talking and TUG can be used to evaluate which patients have a tendency to fall in order to carry out preventive measures. Berg Balance Scale can be used in all patients. Stops Walking When Talking can give additional information if the patient is able to walk. TUG is a possible choice, but fewer patients can perform it. PMID- 16702087 TI - Work participation among persons with traumatic spinal cord injury and meningomyelocele1. AB - OBJECTIVE: To study injury-related and individual factors as predictors of work participation in persons with traumatic and congenital spinal cord injury. DESIGN: Cross-sectional questionnaire study. SUBJECTS: One hundred and eighty-two persons with traumatic spinal cord injury treated in the Spinal Injuries Unit in Sahlgrenska University Hospital, Goteborg, Sweden, and 48 persons with meningomyelocele admitted to the Young Adult Teams in Goteborg, Boras and Skovde, Sweden. METHODS: A structured questionnaire was sent by post. Main outcome variable was participation in work. Logistic regression modelling was used to study the associations between the potential predictors and work participation. RESULTS: Employment rates were 47% in the traumatic spinal cord injury group and 38% in the meningomyelocele group. The presence of other somatic or mental disorder, and neuropathic pain decreased work participation among the men with traumatic spinal cord injury. Among persons with meningomyelocele, better ambulatory status and higher educational level increased work participation. In all groups higher independence in daily activities increased the probability of work participation. According to multivariable modelling carried out for the men with traumatic spinal cord injury, age over 55 years and the presence of mental disorder decreased work participation. CONCLUSION: Our data show that work participation is affected by individual and injury-related factors. Of the latter, many can be affected by rehabilitation. PMID- 16702088 TI - Successful treatment of long-standing post-stroke dysphagia with botulinum toxin and rehabilitation. AB - Cricopharyngeal myotomy is the most common treatment used to restore normal swallowing in patients with persistent (>6 months) cricopharyngeal muscle dysfunction post-stroke. We describe 2 patients whose dysphagia was due to cricopharyngeal muscle over-activity and who significantly improved after a percutaneous botulinum toxin injection in the cricopharyngeal muscle in combination with a rehabilitation treatment (dietary modifications, postural techniques, airflow protection manoeuvres). Swallowing was assessed clinically and by fibreoptic endoscopic evaluation of swallowing and videofluoroscopy; the degree of dysphagia was scored using the penetration-aspiration scale. Two months after the botulinum toxin injection the patients, who were previously fed via percutaneous endoscopic gastrostomy, returned to independent oral feeding and at 6, 12 and 24 months follow-up, both were still able to maintain an adequate oral intake with no signs of aspiration (by videofluoroscopy) or clinical complications. No further botulinum toxin injections or rehabilitation treatments were required. Our findings strongly suggest that even long-standing dysphagia can improve dramatically in selected patients. To the best of our knowledge, there are no other reports with such a long follow-up. PMID- 16702089 TI - Cross-validation of a model for predicting functional status and length of stay in patients with stroke. AB - OBJECTIVE: In a study published in 2002, it was observed that a variable composed by the Functional Independence Measure (FIM) and the trunk control test at admission predicted 66.5% of the FIM at discharge in stroke patients. The objective was to confirm the reproducibility of this predictive model. METHODS: Retrospective study of 245 hemiparetic stroke inpatients of the rehabilitation department. The main variables studied were: trunk control test FIM at admission and compound variable (FIM+trunk control test) as independent variables and FIM at discharge and inpatient rehabilitation length of stay as dependent variables. RESULTS: Correlation between the compound variable and the length of stay was statistically significant (r=0.59), as was its correlation with the total FIM at discharge (r=0.82). The regression analysis predicted 34.3% of the length of stay variability and 66.4% of the total FIM at discharge variability. CONCLUSION: The compound variable is a reliable tool because of its reproducibility in predicting the functional level at hospital discharge in hemiparetic patients. PMID- 16702092 TI - Gesture and speech in maternal input to children with Down's syndrome. AB - BACKGROUND: Despite recent interest in relationships between maternal gesture and speech and communicative development in typically developing (TD) children, little work has examined either speech or gesture in mothers of children with Down's syndrome (DS). AIMS: To compare aspects of speech and gesture production by mothers of children with DS with that of mothers of TD children. METHODS & PROCEDURES: Participants were five mothers of children with DS (mean chronological age (CA) = 47.6 months; mean mental age (MA) = 22.4 months) and five mothers of TD children. To equate for expressive language ability, children in the TD and DS groups were individually matched on the basis of: (1) gender; (2) correspondence between the TD child's chronological age and the DS child's language age; and (3) observed expressive vocabulary size. Each mother-child dyad was videotaped for approximately 30 min during free play. Data analyses focused on: (1) the number and types (speech only, gesture only, mixed) of maternal utterances; (2) the gesture types (deictic, iconic, conventional, emphatic); and (3) for mixed utterances, the structure and the temporal patterning of spoken and gestured components. OUTCOMES & RESULTS: Relative to mothers of TD children, mothers of children with DS produced significantly fewer utterances overall, but the distribution of utterance types did not differ between the two groups. Relative to mothers of TD children, mothers of children with DS used proportionately more deictic gestures and made more frequent use of showing. Mothers of TD children produced more pointing gestures. Finally, mothers of children with DS produced a significantly higher proportion of utterances consisting of a single gesture and a single verbal utterance; in contrast to mothers of TD children, more complex structures (one gesture with multiple verbal utterances, one verbal utterance with multiple gestures) were never observed. Within the category of utterances consisting of a gesture and a single verbal utterance, mothers of children with DS tended to produce gestures that were held throughout the complete verbal utterance, while the gestures of mothers of TD children tended to co-occur with only a portion of the utterance. CONCLUSIONS: The findings suggest that mothers of children with DS adjust their communication to the developmental status of their child. Results are discussed in terms of the role of gesture in maternal communication and in the regulation of mother-child interaction. PMID- 16702093 TI - Phonological deficits in French speaking children with SLI. AB - BACKGROUND: This study investigated the phonological disorders of French-speaking children with specific language impairment (SLI) in production. AIMS: The main goal was to confirm whether children with SLI have limitations in phonological ability as compared with normally developing children matched by mean length of utterance (MLU) and phonemic inventory size. A number of researchers have obtained findings pointing in this direction, but the conclusions have never been tested on French-speaking children. The second goal was to find out whether characteristic features of the French language are reflected in the nature of the children's phonological disorder. METHODS & PROCEDURES: The spontaneous language of 16 children with SLI and 16 control children matched on MLU and phonemic inventory size (normal language development group) were analysed using different measures bearing on utterances, words, syllables and phonemes. In both SLI and NLD groups, the children were distributed into two different subgroups based on their MLU, with controlled phonemic inventory size. OUTCOMES & RESULTS: The results supported a specific limitation in the phonological abilities of French children with SLI, as has already been demonstrated for English, Hebrew, Italian and Spanish-Catalan. However, two unexpected results were also obtained. First, a significant difference between children with SLI and control children could only be found for older children (MLU>3), not for younger children with MLU<3. This was true for all measures. CONCLUSIONS: This finding highlights the importance of having a developmental perspective and needs to be confirmed through a longitudinal study. Second, deficits were much more significant at the phoneme level than at the syllable level. This may be explained by the fact that the pronunciation of syllables in French is very homogenous, making them easier to segment. PMID- 16702094 TI - Real-time language processing in school-age children with specific language impairment. AB - BACKGROUND: School-age children with specific language impairment (SLI) exhibit slower real-time (i.e. immediate) language processing relative to same-age peers and younger, language-matched peers. Results of the few studies that have been done seem to indicate that the slower language processing of children with SLI is due to inefficient higher-order linguistic processing and not to difficulties with more basic acoustic-phonetic processing. However, this claim requires further experimental verification. AIMS: It was investigated whether the real time language processing deficit of children with SLI arises from inferior acoustic-phonetic processing, inefficient linguistic processing, or both poor sensory processing and linguistic processing. If these children's impaired online language processing is due to inferior acoustic-phonetic processing, then their reaction time (RT) for recognizing words presented in list fashion should be significantly longer relative to control children's RT. If, however, their impaired language processing relates to inefficient linguistic processing, then, relative to control children, their RT for word-list-presented words should be comparable and their sentence-embedded word-recognition RT should be significantly longer. METHODS & PROCEDURES: Sixteen school-age children with SLI, 16 age-matched (CA) typically developing children, and 16 receptive-syntax matched (RS) children completed two word-recognition RT tasks. In one task, children monitored word lists for the occurrence of a target word (isolated lexical processing task). In the second task, children monitored simple sentences for a target word (sentence-embedded lexical processing task). In both tasks, children made a timed response immediately upon recognizing the target. OUTCOMES & RESULTS: Children with SLI and CA children showed comparable RT in the isolated lexical processing task and both were faster than RS children. In the sentence processing task, children with SLI were slower at lexical processing than CA and RS children, with CA children demonstrating the fastest processing. CONCLUSIONS: The real-time language processing of children with SLI appears to be attributable to inefficient higher-order linguistic processing operations and not to inferior acoustic-phonetic processing. The slower language processing of children with SLI relative to younger, language-matched children suggests that the language mechanism of children with SLI operates more slowly than what might otherwise be predicted by their linguistic competence. PMID- 16702095 TI - Prosodic and lexical aspects of maternal linguistic input to late-talking toddlers. AB - BACKGROUND: Children who have reached the age of 2 years without having acquired a 50-word vocabulary and/or who use no word combinations are referred to in the literature as 'Late Talkers'. Research has not yet identified the factors that cause slow development of expressive language; in particular, relatively little research has been carried out on the characteristics of the maternal linguistic input received by Late Talkers. In fact, the majority of studies in this area have focused principally on the mothers' semantic, pragmatic and interactive verbal behaviour. AIMS: The objective was to verify whether the input addressed to a group of Late Talkers differs to that addressed to a group of typically developing peers. It was expected that there would be differences in the prosodic and structural/lexical properties of the input, which by creating a 'less than optimal environment' for language acquisition processes, could affect the children's rate of linguistic development. METHODS & PROCEDURES: The spontaneous maternal speech of nine Italian-speaking mothers of late-talking toddlers (LT) at 20 months of age was examined and compared with that of nine mothers of age matched children with typical language development (Late Starters). Data were collected during video-recorded play sessions, which were transcribed and coded based on the structural-lexical and acoustical characteristics of the mothers' speech. Group differences were tested for the following variables: number of utterances per minute, mean number of words per utterance (MLUw), number of nouns per minute, number of different nouns per minute, common nouns as a percentage of overall words, type/token ratio for common nouns, one-word utterances as a percentage of the total number of utterances, one-word utterances formed by a common noun as a percentage of the total number of utterances, mean syllable duration, duration of the final syllable of the utterance, F0 maximum (the highest F0 in the utterance), F0 minimum (the lowest F0 in the utterance) and F0 range (by subtracting F0 min from F0 max). OUTCOMES & RESULTS: The mothers of LT produced common nouns and mark nouns by a pitch peak with a significantly lower frequency than the mothers of the children with typical language development. The mothers of LT also used flat pitch contours more often than the mothers of the typically developing peers. CONCLUSIONS: The study provides evidence that maternal input addressed to Late Talkers can contribute to slowing the process of language acquisition. Based on these findings, additional investigations that examine the contribution of maternal verbal behaviour to the development of language delay are warranted. PMID- 16702096 TI - Language skills in 5-8-year-old children with 22q11 deletion syndrome. AB - BACKGROUND: Language impairment and delayed language onset have been described, although not investigated in detail, in children with 22q11 deletion syndrome. AIMS: To investigate different areas of language: the ability to retell a narrative, phonology, syntax and receptive vocabulary in a group of 5-8-year-old children with 22q11 deletion syndrome regardless of whether or not they had a history of speech and language difficulties. Gender differences were also investigated. METHODS & PROCEDURES: Nineteen consecutively referred children with 22q11 deletion syndrome, ten girls and nine boys, between the ages of 5 and 8 years, participated in the study. The mean full-scale IQ of the group was 78. Six children had an autism spectrum disorder, attention deficit/hyperactivity disorder, or a combination of these. Three different language tests were used: (1) the Bus Story - a test of narrative speech and language; (2) an articulation test including all Swedish phonemes in different positions; and (3) the Peabody Picture Vocabulary Test - Revised (PPVT-R). OUTCOMES & RESULTS: All but two children had an information score in the retelling task of 1 SD below the population mean. A negative correlation between age and the information score implied that the older the children, the more severe the problems. One child had an average sentence length within the normal limits and five children had subordinate clauses within normal limits. A median of 4% of the utterances included grammatical errors. About 50% of the children had a complete consonant inventory. The phonological process analysis implied delayed rather than deviant development. The group had a moderately low score for receptive vocabulary. CONCLUSIONS: Language difficulties in all investigated areas of language were found. It is suggested that speech-language impairment is a common feature of 22q11 deletion syndrome. An implication of these results is that follow-ups of language skills are important not only for pre-school children, but also for school age children and adolescents with 22q11 deletion syndrome. PMID- 16702097 TI - Teaching new words to children with poor existing vocabulary knowledge: a controlled evaluation of the definition and context methods. AB - BACKGROUND: Children who have poor vocabulary knowledge are at risk of wider language weaknesses and reading comprehension difficulties, which will impact upon their educational achievement. The central question addressed in this paper is how best to teach new vocabulary items to these children. AIMS: To investigate the effects of two different methods of teaching vocabulary on both vocabulary knowledge and reading comprehension. METHODS & PROCEDURES: Twenty-four children (aged 7-8 years) with poor existing vocabulary knowledge took part in an intervention study. Half the children were taught new vocabulary items using definitions; the other half were taught a strategy for deriving meanings from written context. Tests of vocabulary knowledge were given before teaching, immediately after teaching and 3 months later. OUTCOMES & RESULTS: Immediately after teaching, both groups had improved equivalently in vocabulary knowledge for the taught words. However, 3 months later, the context group showed significantly better expressive vocabulary knowledge. The context group went on to show significantly better comprehension of text containing a number of the taught words and demonstrated that they could use the newly acquired strategy independently to derive meanings from written context. CONCLUSIONS: The context method developed is effective in increasing vocabulary knowledge and improving reading comprehension in children with poor existing vocabulary knowledge, and this is therefore recommended for use with children who require extra help developing vocabulary and comprehension skills. PMID- 16702098 TI - Dermatophytes: recognizing species of clonal fungi. AB - Now that molecular data have forever changed our perspective on the anthropophilic and zoophilic dermatophyte species, the concepts of these species needs re-evaluation. In this paper, main concepts (morphological, biological (BSC), phylogenetic and genealogical concordance phylogenetic species recognition (GCPSR)) are compared. While in geophilic dermatophytes the application of the BSC works well for species distinction and is supported by molecular data, it is not applicable for the anthropophilic and zoophilic dermatophytes where the majority of species reproduce purely asexually. Also, the application of GCPSR (an operational method to define the limits of species using molecular, multi locus data) is problematic. GCPSR can be applied in recombining fungi even when recombination is infrequent and fungi lack phenotypic sexuality. In truly clonal fungi, however, no incongruities in multi-locus data are found, and thus separation of species may be difficult. In fungi this problem is currently taken to be non-existent, since clonality is supposed to lead to extinction. In the medically relevant, host-associated dermatophytes, however, is reason to suggest that clonal dermatophyte lineages are able to maintain ongoing populations and to follow independent evolutionary trajectories. We distinguish seasonal, short lived and long-lived clonal species. The final goal of a species concept, in the dermatophytes as well as in other fungi, is to provide a taxonomic system that reflects the evolution of the fungal species so that the underlying biological trends elucidated in this way may be brought forward to help to guide the clinician in applying optimal therapy and prophylaxis. The application of the different species concepts may have an enormous impact on the nomenclature of dermatophytes, directly affecting the quality of communications with care providers. PMID- 16702099 TI - The role of the sakA (Hog1) and tcsB (sln1) genes in the oxidant adaptation of Aspergillus fumigatus. AB - The Hog1 MAP kinase pathway regulates stress adaptation in several fungi. To assess its role in stress adaptation in Aspergillus fumigatus, we constructed mutants in genes encoding the sensor histidine kinase (HK) tcsB as well as sakA, which are homologues of the Saccharomyces cerevisiae sln1 and Hog1, respectively. Compared to the wild type strain (Wt), growth of sakA (sakAtriangle up) mutant was reduced, and growth inhibition was increased when H(2)O(2), menadione, or SDS was added to the media. On the other hand, the tcsB mutant (tcsBtriangle up) was similar to the Wt strain in regard to growth and morphology, although a partial sensitivity to SDS was observed. Western blot analysis of Wt and the tcsBtriangle up strains indicated that when stressed with H(2)O(2), phosphorylation of Hog1p still occurs in the mutant. Since in Candida albicans, Hog1 regulates transcription of at least one histidine kinase, we performed RT-PCR of 6 histidine kinase genes as well as the ssk1 and skn7 response regulator genes of A. fumigatus. No significant differences in transcription were observed with the sakAtriangle up when compared to the Wt, indicating that the sakA does not regulate transcription of these genes. Our studies indicate that the A. fumigatus sakA is required for optimal growth of the organism with or without oxidant stress, while tcsB gene is dispensable. PMID- 16702101 TI - Tween 40-based precipitate production observed on modified chromogenic agar and development of biological identification kit for Malassezia species. AB - We developed a simple identification kit for nine species of Malassezia (M. furfur, M. slooffiae, M. sympodialis, M. restricta, M. obtusa, M. globosa, M. pachydermatis, M. dermatis, and M. japonica) based on their biological features. This method utilizes Tween 40-based precipitate production on modified chromogenic agar (CHROMagar) Malassezia medium, growth on specific agars (Sabouraud's dextrose agar, Cremophor EL agar, Tween 60-esculin agar), and catalase reactions. This identification kit was verified with 11 type and reference strains of nine Malassezia species. An additional 26 clinical isolates were also successfully identified using the kit and the results were confirmed by molecular biological analysis. PMID- 16702100 TI - Dysphonia and laryngeal sequelae in paracoccidioidomycosis patients: a morphological and phoniatric study. AB - The larynx is the third most commonly involved organ in paracoccidioidomycosis (PCM). While a few studies have evaluated laryngeal sequelae, there have not been any investigations of voice abnormalities in PCM patients. To evaluate persistent dysphonia and laryngeal lesions, we studied 15 normal subjects and 30 post treatment PCM patients, i.e., 15 with only pulmonary and 15 with both laryngeal and pulmonary involvement. Perceptual and acoustic voice analysis were performed with all patients, while endoscopic studies were also conducted with the 15 laryngeal patients. Voice analysis showed instability by perceptual analysis (P<0.01) in both groups, but more severe dysphonia was noted in the laryngeal group (P<0.01). The dysponia, seen in 66.7% of these patients (dysphonia index < 7.0), was characterized by roughness and breathness. The Dr. Speech (Tiger Electronics) analysis program did not accept five voices from the laryngeal group due to the severe dysphonia. Jitter was elevated in five laryngeal lesion patients. Endoscopy showed that 80% of patients with laryngeal lesion had two or more laryngeal structures involved. Vocal fold alterations were seen in all laryngeal lesion patients, which included involvement of the arythenoids, epiglottis, and vestibular folds. This first functional study of laryngeal sequelae in PCM revealed frequent and severe dysphonia that may have important social consequences for patients. PMID- 16702102 TI - Comparative effects of ertapenem, imipenem, and meropenem on the colonization of the gastrointestinal tract of mice by Candida albicans. AB - We evaluated the effect of three carbapenems on gut colonization of mice by Candida albicans. A total of 150 Crl:CD1 (ICR) BR mice were fed chow containing C. albicans or regular chow. Both groups were subsequently treated either with one carbapenem or with normal saline for 10 days. Stool cultures to determine colonization by C. albicans were performed immediately before, at the end, and one week after the end of treatment. Candida-colonized mice that received carbapenems had substantially higher C. albicans concentrations than control animals fed C. albicans, especially if they received ertapenem. Mice fed regular chow and treated with the study antibiotics or saline did not have Candida in their stools. Candida was not detected in the internal organs of any group of mice. PMID- 16702103 TI - Distribution and antifungal susceptibility of Candida species isolated from different age populations in Taiwan. AB - A total of 3,926 yeast isolates were isolated from 24 hospitals participating in the Taiwan Surveillance of Antimicrobial Resistance of Yeasts (TSARY) from July to September 2002. Candida albicans (69.1%) was the most common species, followed by Candida tropicalis (12.9%), Candida glabrata (8.3%), Candida parapsilosis (2.7%), Candida krusei (0.6%), and others (6.4%). To study the distribution and antifungal susceptibility of Candida species according to sources and patient ages, we have collected and analyzed the clinical data of 861 isolates. Of those 861 isolates, urine was the most common source (40%) followed by sputum (22.1%), blood (13.5%), central venous catheter (5.5%), wound (5.2%), and others (13.7%). With increasing age, we observed a significant increase in the percentage of isolates from urine (P=0.00005) and a parallel reduction from blood (P=0.009). As expected, more elder patients were hospitalized than younger ones (P=0.05). In total, 2.7% and 1.9% of isolates were resistant to amphotericin B and fluconazole, respectively. Antifungal susceptibilities of isolates from different age groups were not significantly different. PMID- 16702104 TI - Isolation, characterization, and disruption of dnr1, the areA/nit-2-like nitrogen regulatory gene of the zoophilic dermatophyte, Microsporum canis. AB - A homolog of the major nitrogen regulatory genes areA from Aspergillus nidulans and nit-2 from Neurospora crassa was isolated from the zoophilic dermatophyte, Microsporum canis. This gene, dnr1, encodes a polypeptide of 761 amino acid residues containing a single zinc-finger DNA-binding domain, which is almost identical in amino acid sequence to the zinc-finger domains of AREA and NIT-2. The functional equivalence of dnr1 to areA was demonstrated by complementation of an areA loss-of-function mutant of A. nidulans with dnr1 cDNA. To further characterize this gene, dnr1 was disrupted by gene replacement based on homologous recombination. Of 100 transformants analyzed, two showed the results expected for replacement of dnr1. The growth properties of the two dnr1(-) mutant strains on various nitrogen sources were examined. Unlike the A. nidulansareA(-) mutant, these dnr1(-) mutants showed significantly reduced growth on ammonia, a preferred nitrogen source for fungi. These mutant strains were also able to utilize various amino acids for growth. In comparison with wild-type M. canis, the two dnr1(-) mutants showed reduced growth on medium containing keratin as the sole nitrogen source. This is the first report describing successful production of targeted gene-disrupted mutants by homologous recombination and their phenotypic analysis in dermatophytes. PMID- 16702105 TI - Isolated endogenous endophthalmitis due to a sporodochial-forming Phialemonium curvatum acquired through intracavernous autoinjections. AB - We report a case of endogenous endophthalmitis due to a sporodochial-forming species of Phialemonium curvatum. The infection led to the enucleation of the affected eye, but there was no evidence of systemic dissemination. The isolated P. curvatum produced aggregates of phialides, many occurring on coils or in verticils, which eventually develop into sporodochia. The initial and post enucleation isolates revealed they were identical to strains of P. curvatum from Israel causing disseminated disease in patients practicing intracavernous autoinjections for the treatment of erectile dysfunction. The reported case had unusual clinical and microbiological features. Despite the route of acquisition and the lack of systemic antifungal therapy, the infection did not spread beyond the eye. The morphology of the phialides aggregates was also unique, and the distinction between Volutella and Acremonium is discussed. This case expands the spectrum of infections due to Phialemonium species, and reveals a novel way of developing fungal endophthalmitis. PMID- 16702106 TI - Clinical and microbiological aspects of candidemia due to Candida parapsilosis in Brazilian tertiary care hospitals. AB - In order to characterize the epidemiology, microbiology and outcome of candidemia due to Candida parapsilosis, we examined a database of 282 episodes of candidemia prospectively collected from four tertiary care hospitals in Sao Paulo, Brazil between March 2002 and February 2003, and compared the characteristics of patients with candidemia due to C. parapsilosis (n=64) with those caused by Candida albicans (n=107). C. parapsilosis candidemia was associated with neutropenia (p=0.005), tunneled central venous catheter (p=0.005) and cancer chemotherapy (p=0.03). By multivariate analysis, candidemia due to C. parapsilosis was associated with the presence of a tunneled central venous catheter (relative risk 3.71, 95% confidence interval 1.28-10.70). Except for a single isolate of C. parapsilosis that exhibited MIC >1 microg/ml to amphotericin B, no resistance was observed in 166 isolates tested against fluconazole, itraconazole, 5-flucytosine and amphotericin B. The caspofungin MIC values of C. parapsilosis isolates were significantly higher than those exhibited by C. albicans isolates (p<0.001). The overall mortality of patients with candidemia due to C. parapsilosis was significantly lower (45% vs. 62%, p=0.03). The association between C. parapsilosis candidemia and a tunneled central venous catheter supports the idea that the main mode of acquisition of C. parapsilosis is from an external source. PMID- 16702107 TI - Differences in beta-glucan levels in culture supernatants of a variety of fungi. AB - (1-->3)-beta-d-glucan is a well known cell wall constituent of fungal isolates that can be detected by assays in vivo and in vitro. Previous studies have shown that different fungal isolates may show different levels of reactivity with an assay for beta glucan. In this study we evaluated the in vitro reactivity of 127 clinical fungal isolates belonging to 40 different genera, with the Glucatell assay. The majority of the fungal isolates released high levels of beta glucan. Beta glucan test reactivity appears to be species-specific and this may reflect the beta glucan content of the organism. PMID- 16702108 TI - Control of a Candida glabrata prosthetic endovascular infection with posaconazole. AB - A 63-year-old man with a history of cirrhosis of the liver developed Candida glabrata fungemia after undergoing transjugular intrahepatic portosystemic shunt (TIPS) placement. Treatment with oral fluconazole was initially effective, but when the patient became neutropenic, subsequent blood cultures grew C. glabrata and a thrombus developed, which partially occluded the stent. Despite treatment with fluconazole, blood cultures remained positive for C. glabrata. Treatment with posaconazole resulted in clinical improvement and the patient had only intermittently positive fungal cultures for 6 weeks. A CT scan showed resolution of the inferior vena cava thrombus. Subsequently, the patient developed hepatocellular carcinoma and hepatic encephalopathy and became noncompliant with posaconazole. Blood cultures again became positive for C. glabrata. The patient died a few weeks after the diagnosis of hepatocellular carcinoma, but the cause of death was believed to be worsening liver dysfunction, not C. glabrata infection. Posaconazole had controlled the infection for about 3 months prior to his death. In conclusion, posaconazole may be a useful option in the management of prosthetic endovascular infections caused by C. glabrata. PMID- 16702109 TI - Cryptococcal parotid involvement: an uncommon localization of Cryptococcus neoformans. AB - We describe a cryptococcal infection localized in the parotid gland of an otherwise healthy 72-year-old woman. The patient presented with a painful, approximately 4.5 cm diameter mass in the anterior region of her right ear. Her symptoms were mild and uncharacteristic. The patient had previously fallen on her face in her garden, causing the loss and breakage of her dentures. Since the soil of the garden contained chicken droppings, it is quite likely that the oral prothesis became contaminated on contacting the soil. The fungus probably entered the parotid gland through the traumatization of the posterior lateral wall of her oral cavity by her broken denture. Numerous intra- and extracellular cryptococcal yeast cells were observed in both histopathological and mycological slide preparations. The yeastlike fungus was recovered in cultures inoculated with tissue collected through three biopsies of her parotid region. The isolates were identified as Cryptococcus neoformans by classical mycology methods and found to be susceptible, in vitro, to fluconazole, amphotericin B and flucytosine. Fluconazole treatment (400 mg/d, for 6 months) was started and the patients facial swelling resolved and the pain significantly reduced within 5 weeks of the initiation of treatment. While fungal infection of the parotid gland have been reported, to our knowledge, this is the first description of a non-disseminated primary parotid infection due to C. neoformans. PMID- 16702110 TI - Honey has an antifungal effect against Candida species. AB - The incidence of Candida infections is escalating worldwide. The serious nature of these infections is compounded by increasing levels of drug resistance. We report that certain honeys have significant antifungal activity against clinical isolates of Candida species. Importantly, the minimum inhibitory concentration of these honeys would be achievable in a clinical setting. PMID- 16702112 TI - Modulation of the growth hormone-insulin-like growth factor (GH-IGF) axis by pharmaceutical, nutraceutical and environmental xenobiotics: an emerging role for xenobiotic-metabolizing enzymes and the transcription factors regulating their expression. A review. AB - The growth hormone-insulin-like growth factor (GH-IGF) axis has gained considerable focus over recent years. One cause of this increased interest is due to a correlation of age-related decline in plasma GH/IGF levels with age-related degenerative processes, and it has led to the prescribing of GH replacement therapy by some practitioners. On the other hand, however, research has also focused on the pro-carcinogenic effects of high GH-IGF levels, providing strong impetus for finding regimes that reduce its activity. Whereas the effects of GH/IGF activity on the action of xenobiotic-metabolizing enzyme systems is reasonably well appreciated, the effects of xenobiotic exposure on the GH-IGF axis has not received substantial review. Relevant xenobiotics are derived from pharmaceutical, nutraceutical and environmental exposure, and many of the mechanisms involved are highly complex in nature, not easily predictable from existing in vitro tests and do not always predict well from in vivo animal models. After a review of the human and animal in vivo and in vitro literature, a framework for considering the different levels of direct and indirect modulation by xenobiotics is developed herein, and areas that still require further investigation are highlighted, i.e. the actions of common endocrine disruptors such as pesticides and phytoestrogens, as well as the role of xenobiotic metabolizing enzymes and the transcription factors regulating their expression. It is anticipated that a fuller appreciation of the existing human paradigms for GH-IGF axis modulation gained through this review may help explain some of the variation in levels of plasma IGF-1 and its binding proteins in the population, aid in the prescription of particular dietary regimens to certain individuals such as those with particular medical conditions, guide the direction of long term drug/nutraceutical safety trials, and stimulate ideas for future research. It also serves to warn athletes that using compounds touted as performance enhancing because they promote short-term GH release could in fact be detrimental to performance in the long-run. PMID- 16702113 TI - Enzymatic characteristics of CYP3A5 and CYP3A4: a comparison of in vitro kinetic and drug-drug interaction patterns. AB - CYP3A4 and CYP3A5 exhibit significant overlap in substrate specificity, but can differ in catalytic activity and regioselectivity. To investigate their characteristics further, the enzymatic reactions of the two CYP3A enzymes were compared using midazolam, nifedipine, testosterone and terfenadine as substrates. Both CYP3A5 and CYP3A4 showed sigmoid and substrate inhibition patterns for testosterone 6beta-hydroxylation and terfenadine t-butylhydroxylation (TFDOH), respectively. In the other reactions, the kinetic model for CYP3A5 was not similar to that for CYP3A4. An inhibition study demonstrated that the interactions between alpha-naphthoflavone (alphaNF) and CYP3A substrates were different for the two CYP3A enzymes. alphaNF stimulated nifedipine oxidation catalysed by CYP3A5, but did not stimulate that catalysed by CYP3A4. alphaNF at less than 32 microM inhibited TFDOH catalysed by CYP3A5, but did not inhibit that catalysed by CYP3A4. These results indicate that CYP3A5 has different enzymatic characteristics from CYP3A4 in some CYP3A catalysed reactions. PMID- 16702114 TI - Metabolism, pharmacokinetics and excretion of a potent tachykinin NK1 receptor antagonist (CP-122,721) in rat: characterization of a novel oxidative pathway. AB - The metabolism, pharmacokinetics and excretion of a potent and selective substance P receptor antagonist, (+)-(2S,3S)-3-(2-methoxy-5 trifluoromethoxybenzlamino)-2-phenylpiperidine, CP-122,721, have been studied in rat following oral administration of a single dose of [14C]CP-122,721. Total recovery of the administered dose was 84.1+/-1.1% for male rat and 80.9+/-2.7% for female rat. Approximately 81% of the administered radioactivity recovered in urine and faeces were excreted in the first 72 h. Absorption of CP-122,721 was rapid in both male and female rat, as indicated by the rapid appearance of radioactivity in plasma. The plasma concentrations of total radioactivity were always much greater than unchanged drug, indicating early formation of metabolites. CP-122,721 t1/2 was 3.1 and 2.2 h for male and female rat, respectively. The plasma concentrations of CP-122,721 reached a peak of 941 and 476 ng ml-1 for male and female rat, respectively, at 0.5 h post-dose. Based on AUC0-tlast, only 1.5% of the circulating radioactivity was attributable to unchanged drug (average of male and female rats) and the balance, approximately 98.5% of the plasma radioactivity was due to metabolites. The major metabolic pathways of CP-122,721 were due to O-demethylation, aromatic hydroxylation and indirect glucuronidation. The minor metabolic pathways included aliphatic oxidation at the piperidine moiety and aliphatic oxidation at the benzylic position of the trifluoromethoxy anisole moiety. In addition, a novel oxidative metabolite resulting from ipso substitution by the oxygen atom and trifluoromethoxy elimination followed by glucuronide conjugation was also identified. PMID- 16702117 TI - NIOSH fills void with surveillance of injuries to youth living on U.S. farms. PMID- 16702115 TI - Urinary excretion of the main metabolites of methamphetamine, including p hydroxymethamphetamine-sulfate and p-hydroxymethamphetamine-glucuronide, in humans and rats. AB - The urinary concentrations of the main metabolites of methamphetamine (MA), specifically p-hydroxymethamphetamine-sulfate (p-OHMA-Sul) and p hydroxymethamphetamine-glucuronide (p-OHMA-Glu), were directly measured in MA users and rats using an optimized LC-ESI MS method. The concentrations of the two conjugates in 50 MA human users' urine ranged from 0.09 to 88.6 microM (0.02-21.7 microg ml-1) for p-OHMA-Sul and from <0.05 to 7.13 microM (<0.02-2.43 microg ml 1) for p-OHMA-Glu; the ratios of sulfate to glucuronide (S/G ratios) ranged from 2.2 to 37.1 (13.8+/-8.1). The results demonstrate that the sulfation is quantitatively more important than glucuronidation for the conjugation of p-OHMA in humans. The urinary concentration time-dependency in two MA users also revealed that the conjugates were mostly excreted in urine within 3 days post intake. In contrast, in rat, almost all of the conjugated p-OHMA (>99%) was excreted as the glucuronide in urine. These findings confirm that a large species variation exists in the conjugation of p-OHMA between humans and rats. PMID- 16702118 TI - Approaching actionable farm safety programs. PMID- 16702119 TI - Hearing loss in migrant agricultural workers. AB - BACKGROUND: Farmers have high rates of hearing loss, yet little is known about the hearing status of migrant agricultural workers. We performed a cross sectional survey to assess the prevalence and impact of hearing loss in this population. METHODS: One hundred fifty migrant and seasonal agricultural workers were surveyed at a series of health fairs held at migrant camps. A bilingual questionnaire included items related to hearing loss risk factors and subjective hearing difficulties. Pure tone audiometry and tympanometry were performed in a mobile testing van. RESULTS: More than half the subjects had some degree of hearing loss at audiometric frequencies between 500 and 6,000 Hz, especially in the higher frequencies. Hispanic males in the sample had significantly greater prevalence of high-frequency hearing loss compared to adults in the national Hispanic Health and Nutrition Examination Survey (HHANES). More than 35% of respondents complained of subjective difficulty hearing or understanding speech, yet no workers reported use of hearing aids. Even after adjusting for measured hearing loss, Hispanic farm workers were more likely than their English- speaking counterparts to complain of difficulty hearing or understanding speech, suggesting that language barriers could worsen the impact of hearing loss. Risk factors for hearing loss included age and abnormal tympanometry. Occupational exposures to noise from tractors and other machinery as well as pesticides were frequently reported, while use of hearing protection was rare. CONCLUSION: Hearing loss is a significant and under-recognized problem in the migrant worker population. Further preventive and treatment efforts are warranted. PMID- 16702120 TI - Injuries to youth living on U.S. farms in 2001 with comparison to 1998. AB - To obtain sustained injury surveillance data for youth on farms, the National Institute for Occupational Safety and Health developed the Childhood Agricultural Injury Survey (CAIS) in collaboration with the U.S. Department of Agriculture. The first CAIS collected data for youth less than 20 years in 1998 through a regionally stratified telephone survey of 50,000 U.S. farm households; a second CAIS for 2001 was conducted using the same methodology. In 2001, there were approximately 1.2 million youth living on U.S. farms. These youth suffered an estimated 19,397 injuries (15.7/1,000 household youth). Approximately 60% (11,571) of the household youth injuries were to males. For all household youth, 10-15 year olds experienced the most injuries (49%, 9,486). In addition to providing estimates of demographics, injuries, and injury rates for household youth from the 2001 CAIS, this article provides a comparison to results from the 1998 CAIS. The number of household youth injuries on farms from 1998 to 2001 decreased by almost 30% (27,321 vs. 19,397). The results of this study show an overall decrease in the injury rate for youth living on the farm from 1998 to 2001 (18.8/1,000 household youth vs. 15.7/1,000 household youth). However, there was a considerable increase in the number of injuries to household females less than 20 years of age during this same time period. There was also an increase in the number of all terrain vehicle (ATV) and horse-related injuries. Continued surveillance is needed to assess if these are significant trends or the result of changing farm demographics. PMID- 16702121 TI - Illnesses related to shank application of metam-sodium, Arvin, California, July 2002. AB - OBJECTIVES: To evaluate the health effects of methyl isothiocyanate (MITC) and other byproducts resulting from the soil-incorporated (shank) application of 25,000 pounds of metam-sodium on July 8, 2002, near the community of Arvin, California. METHODS: Residents in a four-block area were interviewed regarding eye and upper respiratory irritation, non-specific systemic symptoms, and lower respiratory complaints. The distribution of cases was compared to results of Industrial Source Complex (ISC3) air dispersion modeling for the metam-sodium byproduct, methyl isothiocyanate (MITC). The 1-hour 200 ppb no-observed-effect- level (NOEL) and 800 ppb lowest-observed-effect level (LOEL) from a previous human eye irritation study were used to interpret the results of the air modeling estimates. Peak concentrations were compared to the 4-minute NOEL of 600 ppb and the LOEL of 1.9 ppm. RESULTS: Two-hundred-fifty-two cases of irritant, non specific systemic, and respiratory symptoms were associated with the metam-sodium application. These included 178 community residents or visitors and 74 employees of a carrot packing operation located in the affected neighborhood of Arvin. The most severe reported illness occurred in a community visitor with a history of pre-existing pulmonary disease, who was hospitalized for a week with respiratory distress. ISC3 Modeling indicated 1-hour MITC concentrations in the affected community ranged from 0.8-1.0 ppm, in the range of the LOEL, with peak concentrations between 2.4 and 3.2 ppm. CONCLUSION: Estimated MITC concentrations during the episode exceeded both the 4-minute NOEL and 1-hourNOELby approximately four fold. The high concentrations of MITC present in the affected neighborhood may have been partially attributable to failure to immediately complete a required post-application water-treatment on 15 of the 100 treated acres. However, because of the limited area involved, the violation was unlikely to have accounted for the entire incident. Similar episodes may occur when metam-sodium fumigants are used adjacent to other rural communities. PMID- 16702123 TI - Health effects from breathing air near CAFOs for feeder cattle or hogs. AB - There is concern that livestock operations for fattening cattle and raising hogs known as concentrated animal feeding operations (CAFOs) release substances into the air that have negative effects on the health of persons living nearby. These substances include dust containing endotoxin and other microbial products as well as ammonia, hydrogen sulfide and a variety of volatile organic compounds. Odors from these farms are considered offensive by some neighbors. A variety of medical complaints are reported to be more common in those people who live near CAFOs for raising hogs than in people without this exposure. Respiratory health effects, including symptoms of pulmonary disease and lung function test result abnormalities, have been described in workers employed in CAFOs where hogs are raised. Health effects after inhalation exposure of neighbors to substances released into the ambient air from these farms is less well characterized. It must be noted that CAFO workers may differ from neighbors in terms of their exposures and general health status. The presence of dust and other substances from cattle feedlots also causes some neighbors to voice concerns about the impact on their health but this exposure has been studied less extensively than exposure to substances released from CAFOs where hogs are raised. Further research needs to be done to look for measurable health effects attributable to living near all CAFOs in order to better understand the impact of these farms. PMID- 16702122 TI - Persistent neuropathy and hyperkeratosis from distant arsenic exposure. AB - The purpose of this case series is to assess long-term sequelae of arsenic exposure in a cohort acutely exposed to arsenic in drinking water from a well dug into a landfill containing arsenical pesticides. Ten of the 13 individuals (or next of kin) in the initial study agreed to participate in the follow-up study. Next of kin provided questionnaire data and released medical information on the three individuals who had died. The remaining seven cohort members were assessed by an interview, questionnaire, detailed physical examination and sensory nerve testing. Available medical records were obtained and reviewed. Sensory testing was performed using an automated electrodiagnostic sensory Nerve Conduction Threshold (sNCT) evaluation. Sensory complaints and electrodiagnostic findings consistent with polyneuropathy were found in a minority (3/7) of subjects 28 years after an acute toxic arsenic exposure. Two of the seven patients examined (1 of 3 with neuropathic findings) also had hyperkeratotic lesions consistent with arsenic toxicity and one of the patients had hyperpigmentation on their lower extremities possibly consistent with arsenic toxicity. PMID- 16702124 TI - "Safety for agricultural educators": evaluation of an intervention to enhance awareness of agricultural hazards. AB - Safety for Agricultural Educators (SAGE) is a two-sided fact sheet on agricultural hazards that has been distributed by the New York Center for Agricultural Medicine and Health (NYCAMH) to Northeastern high school agriculture educators over the past six years. It was intended to supplement and stimulate the teaching of safety principles in agricultural education classes and thus enhance the awareness and knowledge of young people working on farms. This paper describes an evaluation of SAGE's success at achieving these goals. Postal surveys from 29 teachers who had not previously received SAGE gathered data on class demographics and the amount of class time devoted to a variety of agricultural topics including tractor operation and agricultural health and safety. Teachers were asked to assess their students' awareness of safety and health issues and also to estimate the number of students working in the farm environment. Following a year of SAGE mailings, the same information was gathered from the 24/29 post-SAGE responders. Comparison of pre-post responses from the 24 matched responding educators showed: limited increase in the teachers' knowledge of basic agricultural safety information; a decline in the amount of class time devoted to health and safety; a marginal increase in estimated student awareness of health and safety and very little evidence of teacher awareness of SAGE as a safety resource. Most notable was the fact that only 8% of all students were currently working on a farm and teachers estimated that only 6% would be working on farms in five years. PMID- 16702125 TI - Survey of health needs and concerns of rural Pennsylvanians. AB - INTRODUCTION: This survey was undertaken to determine the most important health and safety needs of farmers and rural citizens of Pennsylvania. METHODS: In June of 2004, a one-page questionnaire was distributed by mail to 32,700 members of the Pennsylvania Farm Bureau as an attachment to their monthly newsletter. RESULTS: The lack of affordable health care insurance was overwhelmingly cited as the most important unmet health care need (73%). No other issue was chosen by more than 10 percent of respondents. This lack of affordable health insurance was also cited as the most important health or safety problem to be addressed in the respondent's communities (71%). When asked the most important health or safety problem affecting them as individuals or their community, the respondents chose "no major problem or need" as the most frequent response (46%). The issue of affordable health insurance was not an option for this question. The predominant concern regarding the lack of affordable health insurance was consistent across all regions of the Commonwealth and for farmers as well as residents of all community sizes sampled. DISCUSSION: This study would indicate that the lack of affordable health insurance is the preeminent health and safety issue for many Pennsylvania farmers and rural citizens. PMID- 16702131 TI - Psychiatric and pain characteristics of prescription drug abusers entering drug rehabilitation. AB - There has been intense interest in the problem of prescription drug abuse on the parts of health professionals, law enforcement, the media, and the general public. Clinicians not only need to know how to assess risk but also what drugs are being diverted most in their region. We conducted a prospective survey of prescription drug abusers entering a treatment facility in central Kentucky. Participants (n = 109) were enrolled and completed a structured clinical interview and prescription drug abuse survey. The prescription drug abusers assessed in the study had a mean age of 30.95 years (SD = 10.21), were comprised of 75 men (69%) and 34 women (31%), and were mostly Caucasian (98%). The majority (84%) stated that they had legitimately been given a prescription for opioids for pain at some point from a physician and 61% reported chronic pain concerns. The most commonly abused drugs were hydrocodone-containing formulations (78%) and oxycodone-containing products (69%), while products containing methadone (23%) or fentanyl (7%) were abused much less frequently. Most respondents (91%) stated that they had purchased prescription opioids from a street dealer at least once and the majority (80%) had altered the delivery system of the prescription drug by chewing, snorting, or using i.v. administration. Implications for pain management are discussed, focusing on the need for clinicians treating chronic pain to more thoroughly assess patients for their risk of abuse and addiction before starting an opioid regimen. PMID- 16702132 TI - What doctors know about cancer pain management: an exploratory study in Sarawak, Malaysia. AB - Effective cancer pain management is influenced by the attitudes and knowledge of treating physicians. A survey was conducted among the total population of government hospital doctors of Sarawak to study the barriers to cancer pain management. Two hundred and fifty-three respondents (83%) completed the survey. The study results highlight that knowledge about cancer pain management was low and barriers to morphine prescription were high. A majority of doctors were deterred from using morphine because of fear of addiction (36.5%) and respiratory depression (53.1%). Only 16.2% of the doctors chose the oral mode of administration to treat pain, furthermore 25% prescribed morphine on "PRN" basis. Doctors with undergraduate study in oncology consistently answered better suggesting that the situation can be improved by education. This study showed that barriers to morphine prescription and knowledge deficit amongst government doctors in Sarawak are strong but similar to those reported in western countries few years ago. PMID- 16702133 TI - Dexmedetomidine: a novel analgesic with palliative medicine potential. AB - Dexmedetomidine has gained popularity in anesthesia and critical care for use in deep sedation and analgesia due to a combination of its efficacy and safety compared with other available agents (e.g., opioids, benzodiazepines, propofol) conventionally used in these settings. This brief review is meant to introduce this unique agent to the palliative care field, as dexmedetomidine may hold promise for patients in hospice and palliative care settings whose symptoms are refractory to usual therapies. [Be sure to be clear in the abstract that more studies are warranted and its role is not well defined and is complicated by significant drug interactions, invasive i.v. route and has a significant side effect profile.] PMID- 16702134 TI - A prototypical gateway to the World Wide Web: Making sense of pain relief. AB - The World Wide Web site entitled Making Sense of Pain Relief is described. This site, intended for consumers, was created by the American Chronic Pain Association, American Pain Foundation and National Pain Foundation. Other websites directly linked to this site are listed. PMID- 16702135 TI - Evidence-based pain management and palliative care in issue four for 2005 of The Cochrane Library. AB - The Cochrane Library of Systematic Reviews is published quarterly. It now contains 2524 complete reviews, 1589 protocols of reviews and 5585 one page summaries of systematic reviews published in the general medical literature. In addition there are citations of 463,763 randomized controlled trials, 20 methodology reviews and 7484 cited papers in the Cochrane methodology register. The health technology assessment database contains 4733 citations. This edition of the Library contains 89 new reviews of which 6 have potential relevance for practitioners in pain and palliative medicine. PMID- 16702136 TI - Botox for back pain and "regular" versus migraine headaches. AB - This feature presents information for patients in a question and answer format. It is written to simulate actual questions that many pain patients ask and to provide answers in a context and language that most pain patients will comprehend. Issues addressed in this issue are the use of botulinum toxin to treat back pain and differences between tension-type and migraine headaches. PMID- 16702137 TI - Existential pain: impressions from an American Indian reservation. AB - The existential suffering of members of the Havasupai Indian tribe are described by a physician-anthropologist who spent some time serving as an interim medical provider for the tribe. Pain is often described as having both physical and emotional dimensions. In this paper, the existential dimension is explored through a narrative of the author's experiences. PMID- 16702138 TI - Amyotrophic lateral sclerosis prognosis, Alzheimer's disease and end-of-life care, anxiety and cancer, cirrhosis as a terminal illness. AB - This feature is based on actual questions and answers received and responded to by the Hospice Foundation of America (HFA). This is a service provided for families and support group members of patients with advanced disease by William M. Lamers, MD, HFA Medical Consultant a member of this Journal's Editorial Board. In this issue, queries and responses are presented addressing the prognosis for a patient with amyotrophic lateral sclerosis, end-of-life care for a patient with advanced Alzheimer's disease, anxiety in cancer and cirrhosis as a life ending disease. PMID- 16702139 TI - Controlled prescription drug abuse at epidemic level. AB - In July 2005, The National Center on Addiction and Substance Abuse (CASA) at Columbia University announced the results of a study indicating that the number of Americans who abuse controlled prescription drugs has nearly doubled from 7.8 million to 15.1 million from 1992 to 2003 and abuse among teens has more than tripled during that time. A summary of that study and information about the full report are presented. PMID- 16702140 TI - End-of-life care: questions and answers. AB - When cancer is not curable, the focus of care shifts from cure to care. Patients and family members then have many questions about end-of-life care. The following information provided by the National Cancer Institute addresses life expectancy, when caregivers should ask for help, how caregivers can be attentive to patients' emotional needs, signs of dying and death, what to do after death, and where additional resources on end-of-life care can be found. PMID- 16702143 TI - Nonprescription analgesic use for minor arthritis pain increasing. PMID- 16702144 TI - Sexual abuse and subsequent suicidal behaviour. Exacerbating factors and implications for recovery. AB - Suicidal behaviour is a cause for concern among many western countries; in general, it is most common among young women. This research used qualitative methods to explore the narratives of 24 women, to contextualise their insights, and to examine the meanings of events leading to and implicated in the recovery from suicidal behaviour. The research confirms sexual abuse as a common precursor to suicidal behaviour; several women asserted that they would not have attempted suicide if they did not have a sexual abuse history. The women noted that the effects of sexual abuse were exacerbated by problems with disclosure, linking to issues of control, with implications for intervention and recovery. PMID- 16702146 TI - Factors that predict how women label their own childhood sexual abuse. AB - Despite the psychological impact of child sexual abuse, many victims do not acknowledge that their experiences were "abuse". This study sought to identify factors that predict how women label their own experiences of childhood sexual abuse. This cross-sectional study was conducted in a family medicine clinic with adult female patients. Subjects completed structured interviews about their childhood environment and their sexual abuse history. Logistic regression analysis showed that labeling of abuse was dependent upon intercourse (beta = 7.43, p = .006), the frequency of abuse by the first perpetrator (beta = 5.08, p = .024), and paternal overprotection (beta = 6.69, p = .010). Findings suggest that the severity of abusive acts is most important and an over-protective father may enhance the victim's acknowledgment that sexual touching is abusive. PMID- 16702145 TI - The process of coping with domestic violence in adult survivors of childhood sexual abuse. AB - Research suggests that the use of disengaged or avoidant strategies to cope with interpersonal violence contributes to the development of depressive symptoms and other psychological difficulties. Survivors of childhood sexual abuse (CSA) who are exposed to subsequent episodes of abuse may be more likely to rely on disengaged coping strategies, placing them at elevated risk of psychological symptomatology. In this study, we explored the interrelationships between coping, depression, and self-esteem in an ethnically diverse sample of domestic violence survivors (N = 219) with and without a history of CSA. As predicted, CSA survivors (n = 86) reported significantly greater use of disengaged coping strategies (wishful thinking, self-criticism, and social withdrawal) than non-CSA survivors (n = 133). As hypothesized, both a CSA history and the use of disengaged coping significantly predicted higher levels of depression and lower self- esteem. Clinical implications of the findings are discussed. PMID- 16702148 TI - "Speaking of mothers. . ."How does the literature portray mothers who have a history of child sexual abuse? AB - The tendency to cast mothers in a negative light has featured in much of the literature researching child sexual abuse. More recently however, this trend has become increasingly evident in any number of empirical discussions focusing on mothers who themselves have a history of child sexual abuse. This article presents a detailed thematic analysis of the ways in which the literature presents these mothers, particularly concentrating on research examining their parenting post disclosure of their child's sexual abuse. A critical reading of the literature supports commentary on methodological problems in research design as well as assumptions made about mothering, the nature of support, and the focus on the effects of childhood sexual abuse to the exclusion of the influence of other childhood experiences and difficulties. PMID- 16702149 TI - Intergenerational transmission of sexual abuse? Motherhood in the shadow of incest. AB - The aim of the study was to understand why the dynamic of sexual abuse is perpetuated across successive generations. A qualitative analysis was conducted on therapy session transcripts and diaries written during the therapy of 24 mothers who were survivors of incest, and whose children were the victims of incest. Four types of mothers were defined: the Unaware mother, characterized by a complete lack of cognitive knowledge of the sexual abuse occurring in her home; the Unwitting Accomplice, characterized by latent cooperation with the sexual abuse perpetrated by her husband; the Enabler, characterized by overtly or covertly encouraging her spouse in the raping of her daughter; and the Common Fate mother, characterized by sharing a common fate with her daughters. PMID- 16702151 TI - Reactions to internal and external criticism of outgroups: social convention in the intergroup sensitivity effect. AB - Recent research has documented the intergroup sensitivity effect (ISE) whereby people respond more favorably to internal versus external criticism of their group. The present studies examine the reactions of bystanders who do not belong to the criticized group and whose reactions are therefore more likely to be informed by social conventions than by defensiveness. Studies 1 and 2 presented British participants with criticisms of Australians, manipulating their ostensible source. These British bystanders exhibited the ISE, responding more favorably to the speaker and comments when the critic was Australian rather than non-Australian. These responses were driven by the perceived motives of speakers rather than their level of experience with the group (Study 2). Study 3 provides direct evidence that internal criticism is more conventionally acceptable than is external criticism. PMID- 16702150 TI - Non-abusive mothers of sexually abused children. The role of rumination in maternal outcomes. AB - This study of 125 mothers examined the role of rumination in maternal emotional and behavioral outcomes subsequent to discovery of the sexual abuse of their children. Abuse severity, a maternal history of child abuse experiences, and life hassles were examined as predictors of negative outcomes. The central finding was that these factors, many of which are not controllable, were less likely to predict poor maternal outcomes than was rumination, a cognitive process that may be alterable. Rumination was the strongest correlate with each outcome. Further, the effects of most predictors on outcomes in this study were mediated by a ruminative cognitive style. Abuse severity, income, ethnicity, and education lacked significant relationships with maternal outcomes. This study asserts that rumination is a central component for understanding maternal outcomes in the post discovery phase of sexual abuse cases. It recommends that rumination be routinely assessed both for research and treatment purposes. PMID- 16702152 TI - Striving for success in outgroup settings: effects of contextually emphasizing ingroup dimensions on stigmatized group members' social identity and performance styles. AB - For members of stigmatized groups, being confronted with highstatus outgroup members threatens social identity and undermines performance on status-relevant dimensions. Two experiments examined whether the negative effects of outgroup contexts are alleviated when value is expressed for a dimension on which the stigmatized ingroup excels. Specifically, the authors assessed whether ingroup versus outgroup context and contextual value for ingroup dimensions affects group members' reactions to failure on status-relevant dimensions and subsequent performance. Experiment 1 showed that in comparison to ingroup contexts, outgroup contexts induce stigmatized group members to protect social identity and to feel more agitated following negative performance feedback. Experiment 2 showed that when others in the context emphasize the importance of a dimension on which the ingroup excels, the negative effects of outgroup contexts are alleviated, stigmatized group members feel more cheerful concerning an upcoming task, and task performance is characterized by a focus on success. PMID- 16702153 TI - How to bite your tongue without blowing your top: implicit evaluation of emotion regulation predicts affective responding to anger provocation. AB - People frequently have to control their emotions to function in life. However, mounting evidence suggests that deliberate emotion regulation often is costly. This presents a dilemma: Is it better to let emotions go or to pay the price of exerting costly control? In two studies, the authors explore whether emotion regulatory processes associated with implicit positive evaluation of emotion regulation might provide the benefits of successful emotion regulation without the costs. In Study 1, the authors introduce a measure of implicit evaluation of emotion regulation (ER-IAT). Study 2 examined whether this measure is associated with actual emotional responses to an anger provocation. It was found that greater ER-IAT scores were associated with lesser anger experience, fewer negative thoughts, lessened self-reported effortful emotion regulation, and an adaptive pattern of cardiovascular responding. These findings suggest that implicit positive evaluation of emotion regulation is associated with successful, automatic, and physiologically adaptive down-regulation of anger. PMID- 16702154 TI - Time, uncertainty, and individual differences in decisions to cooperate in resource dilemmas. AB - Many environmental problems are resource dilemmas that contain two components social and temporal-that encourage individuals not to act in the best interest of the group. Most research has focused on the social component. The authors examined the importance of the temporal component in two resource dilemma tasks. Participants (N = 112) reported their willingness to limit resource consumption in response to hypothetical dilemmas and forfeited extra credit points in response to a real dilemma. Cooperation rates were higher when the temporal dilemma was diminished, when uncertainty about environmental impacts was low, and for women. Individuals high in both proenvironmentalism and consideration of future consequences sustained high levels of cooperation even in the face of strong temporal dilemmas. PMID- 16702155 TI - Sex and power in the academy: modeling sexual harassment in the lives of college women. AB - The authors build an integrated model of the process by which academic sexual harassment undermines women's well-being; also examined is harasser power as a potential moderator of this process. Data from 1,455 college women suggest that sexual harassment experiences are associated with increased psychological distress, which then relates to lower academic satisfaction, greater physical illness, and greater disordered eating. The cumulative effect is greater disengagement from the academic environment, which in turn relates to performance decline (i.e., lower grades). Regardless of how frequently the harassment occurred, academic satisfaction was lower when the harassment came from higher status individuals (i.e., faculty, staff, or administrators). At the same time, harassment was equally detrimental to mental health, regardless of who perpetrated it. The article concludes with implications for theory, research, and intervention. PMID- 16702156 TI - Driven to tears or to joy: response dominance and trait-based predictions. AB - The present studies reinvigorate the construct of traitedness from a cognitive perspective. Tendencies toward habit (vs. flexibility) were assessed by examining response dominance performance within choice reaction time tasks. Consistent with the idea that response dominance reflects a tendency toward habitual modes of thought and action, three studies involving 428 undergraduates revealed that trait-outcome and test-retest correlations were higher among individuals higher in response dominance. In Studies 1 and 2, such trait-consistency effects took the form of stronger relations between extraversion and neuroticism, on one hand, and mood states and behavior, on the other. In Study 3, such tendencies took the form of higher test-retest correlations related to daily experiences of mood states, somatic symptoms, and life satisfaction. Together, the studies reveal a consistent effect of response dominance on trait-like consistency and raise some important issues for future studies of the traitedness construct. PMID- 16702157 TI - The mixed blessings of self-knowledge in behavioral prediction: enhanced discrimination but exacerbated bias. AB - Four experiments demonstrate that self-knowledge provides a mixed blessing in behavioral prediction, depending on how accuracy is measured. Compared with predictions of others, self-knowledge tends to decrease overall accuracy by increasing bias (the mean difference between predicted behavior and reality) but tends to increase overall accuracy by also enhancing discrimination (the correlation between predicted behavior and reality). Overall, participants' self predictions overestimated the likelihood that they would engage in desirable behaviors (bias), whereas peer predictions were relatively unbiased. However, self-predictions also were more strongly correlated with individual differences in actual behavior (discrimination) than were peer predictions. Discussion addresses the costs and benefits of self-knowledge in behavioral prediction and the broader implications of measuring judgmental accuracy of judgment in terms of bias versus discrimination. PMID- 16702158 TI - The positive and negative framing of affirmative action: a group dominance perspective. AB - Using a sample of 328 White, Latino, and Black Los Angeles County adults, the authors examined the tendency to employ various affirmative action "frames" (e.g., affirmative action as a "tie-breaking" device or as a quota-based policy). All three groups agreed about which frames cast affirmative action in a positive light and which cast it in a negative light. Although minorities had a tendency to frame affirmative action in terms that most people find morally acceptable, Whites had a tendency to frame affirmative action in terms most people find unacceptable. In addition, compared to minorities, Whites were less supportive of affirmative action regardless of how it was framed. LISREL modeling also was employed to test two competing models regarding predictors of the tendency to use frames that one personally finds to be relatively negative versus positive. Consistent with the expectations of social dominance theory and a motivated cognition perspective, the authors found that social dominance orientation (SDO) had significant net direct and indirect effects on one's framing of affirmative action. PMID- 16702159 TI - The "ripple effect": cultural differences in perceptions of the consequences of events. AB - Previous research has demonstrated that people from East Asian cultural backgrounds make broader, more complex causal attributions than do people from Western cultural backgrounds. In the current research, the authors hypothesized that East Asians also would be aware of a broader, more complex distribution of consequences of events. Four studies assessed cultural differences in perceptions of the consequences of (a) a shot in a game of pool, (b) an area being converted into a national park, (c) a chief executive officer firing employees, and (d) a car accident. Across all four studies, compared to participants from Western cultural backgrounds, participants from East Asian cultural backgrounds were more aware of the indirect, distal consequences of events. This pattern occurred on a variety of measures, including spontaneously generated consequences, estimations of an event's impact on subsequent events, perceived responsibility, and predicted affective reactions. Implications for our understanding of cross cultural psychology and social perception are discussed. PMID- 16702160 TI - Differential effects of right wing authoritarianism and social dominance orientation on outgroup attitudes and their mediation by threat from and competitiveness to outgroups. AB - A dual-process model of individual differences in prejudice proneness proposes that Right Wing Authoritarianism (RWA) and Social Dominance Orientation (SDO) will influence prejudice against particular outgroups through different motivational mechanisms. RWA should cause negative attitudes toward groups seen as threatening social control, order, cohesion, and stability, such as deviant groups, and negativity toward these groups should be mediated through perceived threat from them. SDO should cause negative attitudes toward groups that activate competitiveness over relative dominance and superiority, such as socially subordinate groups low in power and status, and negativity toward these groups should be mediated through competitiveness toward them. Findings from four student samples that assessed attitudes toward seven social groups selected as likely to vary systematically in social threat and social subordination supported these predictions. The findings have implications for reconciling intergroup and individual difference explanations of prejudice and for interventions to reduce prejudice. PMID- 16702161 TI - No man is an island: the need to belong and dismissing avoidant attachment style. AB - The need to belong theory proposes that all human beings need social connections. However, dismissive avoidant individuals claim to be comfortable without close relationships and appear to be indifferent to how other people think of them. The current studies examined the association between dismissing avoidant attachment and the desire to feel accepted by others. In Study 1, high-dismissive participants reported experiencing higher than average levels of positive affect and state self-esteem after learning that other participants accepted them. In Study 2, high-dismissive participants felt better about themselves and experienced higher levels of positive affect after learning that in the future they would be successful in interpersonal relative to independent domains or controls. The results of these studies suggest that dismissive avoidants do not represent a counterexample to the hypothesis that all human beings have a fundamental need and desire to belong. PMID- 16702162 TI - Phase I study of single-dose oxaliplatin in Japanese patients with malignant tumors. AB - BACKGROUND: Oxaliplatin, a platinum compound, has been commonly used around the world for treating advanced colorectal cancer. The generally recommended dose and schedule of oxaliplatin monotherapy is 130 mg/m(2) every 3 weeks. This trial was conducted to evaluate the safety and pharmacokinetics of oxaliplatin monotherapy in Japanese patients with solid tumors. METHODS: Oxaliplatin was administered as a 2-h intravenous infusion every 3 weeks at a dose of 90 and 130 mg/m(2). Blood was collected to determine the total platinum and the ultrafiltrate platinum concentrations in plasma in all cycles. RESULTS: Nine patients were enrolled; three were given oxaliplatin monotherapy at 90 mg/m(2) and six received 130 mg/m(2). All tumors were colorectal cancer. The major adverse reactions included myelosuppressive, neurological and gastrointestinal toxicities, although most were grades 1 and 2 at both dose levels. Peripheral sensory neuropathy of without movement disturbance (grade 1 or 2) was observed in all patients at both dose levels. The 130 mg/m(2) dose level was not found to be the maximum tolerated dose, but was judged to be the recommended dose. No objective responses were seen and five cases of no change were observed. A bi-exponential open model best described the disappearance of platinum in the plasma, and a tri-exponential open model best described the disappearance of ultrafilterable platinum in the plasma at both dose levels. No racial difference was suggested in the pharmacokinetics of oxaliplatin. CONCLUSIONS: The oxaliplatin monotherapy dose schedule of 130 mg/m(2) every 3 weeks, recommended worldwide, is acceptable for Japanese patients. PMID- 16702164 TI - T1 neuroendocrine carcinoma of anal canal after transanal resection for intramucosal adenocarcinoma. AB - Neuroendocrine carcinomas of the anal canal are rare, representing 1% of malignant tumors of the anal canal. This tumor behaves aggressively and leads to poor outcomes. The majority of tumors are found with distant metastases. We describe the case of a 63-year-old female with T1 neuroendocrine carcinoma of the anal canal arising from the site of a previous transanal excision performed 13 months earlier for intramucosal adenocarcinoma of the anal canal. The patient did not have any distant metastases on preoperative computed tomography and magnetic resonance imaging. She underwent abdominoperineal resection after the initial diagnostic transanal excision of the neuroendocrine carcinoma, which had shown submucosal invasion. No lymph node metastasis was found in pathological examination. In this case, it is likely that the neuroendocrine tumor, which infiltrated into the submucosal layer with venous invasion, had developed over the intervening 13 months following the original transanal excision of the adenocarcinoma. PMID- 16702163 TI - Interstitial shadow on chest CT is associated with the onset of interstitial lung disease caused by chemotherapeutic drugs. AB - OBJECTIVE: Pretreatment computerized tomography (CT) films of the chest was studied to clarify the influence of interstitial shadow on developing interstitial lung disease (ILD). METHODS: Eligible patients were those lung cancer patients who started to receive first-line chemotherapy between October 2001 and March 2004. Patients who received thoracic radiotherapy to the primary lesion, mediastinum, spinal or rib metastases were excluded. We reviewed pretreatment conventional CT and plain X-ray films of the chest. Ground-glass opacity, consolidation or reticular shadow without segmental distribution was defined as interstitial shadow, with this event being graded as mild, moderate or severe. If interstitial shadow was detected on CT films of the chest, but not via plain chest X-ray, it was graded as mild. Patients developing ILD were identified from medial records. RESULTS: A total of 502 patients were eligible. Mild, moderate and severe interstitial shadow was identified in 7, 8 and 5% of patients, respectively. A total of 188 patients (37%) received tyrosine kinase inhibitor (TKI) treatment, namely gefitinib or erlotinib. Twenty-six patients (5.2%) developed ILD either during or after chemotherapy. Multivariate analyses revealed that interstitial shadow on CT films of the chest and treatment history with TKI were associated with the onset of ILD. CONCLUSIONS: It is recommended that patients with interstitial shadow on chest CT are excluded from future clinical trials until this issue is further clarified, as it is anticipated that use of chemotherapeutic agents frequently mediate onset of ILD in this context. PMID- 16702165 TI - JunD/AP-1 and STAT3 are the major enhancer molecules for high Bcl6 expression in germinal center B cells. AB - The Bcl6 proto-oncogene, which encodes a transcriptional repressor, is ubiquitously expressed and predominantly in germinal center (GC) B cells. Although the promoter region of the human Bcl6 gene has been reported, enhancer molecules for its high expression in GC B cells were largely unknown. Here we show that transcriptional start sites of the murine Bcl6 gene were different from the reported human one. DNA sequence around the new promoter region is highly conserved between mice and humans and has no canonical TATA or CCAAT box. Two AP 1-binding elements in the promoter region were the major enhancer elements in GC derived B lymphoma cells, and JunD/AP-1 was detected in GC B cells. In addition, we identified the silencer region with three Bcl6-binding elements around the start site. Bcl6 bound to the silencer elements and its over-expression repressed the promoter activity through the elements. Activated STAT factors (STATs), especially activated STAT3, also bound to the silencer elements in GC B cells and competed with Bcl6 for the binding, suggesting that JunD/AP-1 and activated STATs drive high Bcl6 expression in GC B cells. Since stimulation of splenic B cells with IL-4 or IL-21 induced high Bcl6 expression with induction of junD and activation of STATs, these cytokines may be inducers for its high expression in GC B cells. However, IL-21 but not IL-4 stimulation activated STAT3 in splenic B cells. Thus, IL-21 may be a major inducer for high Bcl6 expression in GC B cells. PMID- 16702166 TI - Defective cell cycle induction by IL-2 in naive T-cells antigen stimulated in the presence of refractory T-lymphocytes. AB - CD4+ T cells enter a transient refractory period after stimulation. Upon re stimulation the refractory cells produce little IL-2 and show diminished proliferation. We previously demonstrated that refractory T cells can also, like anergic and CD4+CD25+ regulatory T cells, suppress in trans the proliferation of antigen-stimulated naive T cells. The suppressed T cells up-regulate high affinity IL-2R but do not produce IL-2. This IL-2 deficit could potentially explain the proliferation failure, but does not appear to do so. Supplementation of refractory-naive co-cultures with exogenous IL-2 fails to alleviate both the proliferation suppression and IL-2 production defects. This does not result from a failure of IL-2 to stimulate its receptor. Proximal IL-2 signaling into suppressed T cells through STAT5 and Akt is intact. However, refractory cell-co cultured T cells fail to up-regulate cyclins and c-myc and incompletely down regulate p27kip1 in response to IL-2, and the downstream consequences of this signaling are therefore dissociated. IL-2 signaling is not fully disabled as IL-2 up-regulates the anti-apoptotic protein Bcl-xL to control levels. This up regulation correlates with enhanced survival of refractory cell-co-cultured T cells placed in IL-2 when compared with cells cultured without IL-2. Thus, refractory T cells are able to suppress naive T-cell proliferative responses in part by blocking both IL-2 production and the mitogenic but not anti-apoptotic effects of IL-2. These results have implications for how activation-refractory T cells may influence nascent immune responses. PMID- 16702167 TI - Impact of effector cell differentiation on CD4+ T cells that evade negative selection by a self-peptide. AB - We have used a transgenic mouse system to examine how differing reactivities of TCRs expressed by naive versus effector cells can shape the functional potential of autoreactive CD4+ T cells. Transgenic mice expressing TCRs that exhibit either high (TS1) or low [TS1(SW)] reactivity toward the I-Ed-restricted determinant S1 from the influenza virus PR8 hemagglutinin (HA) were mated with transgenic mice expressing HA under the control of different promoters. HACII mice express HA driven by an MHC class II promoter, and both the TS1 and TS1(SW) TCRs underwent substantial deletion in this background. HA104 mice express HA driven by an SV40 promoter, and the highly reactive TS1 TCR was substantially deleted. By contrast, the less reactive TS1(SW) TCR underwent little or no deletion in TS1(SW) x HA104 mice, although CD5 up-regulation indicated that they had interacted with the S1 self-peptide. In adoptive transfer studies, naive CD4+ T cells expressing the TS1(SW) TCR failed to proliferate in response to the S1 peptide in HA104 mice, and were inefficient at providing help for HA-specific antibody responses. However, effector CD4+ T cells generated from TS1(SW) x HA104 mice acquired the ability to proliferate in response to the S1 peptide in HA104 mice, and were as efficient as CD4+ T cells expressing the high reactivity TS1 TCR in helping HA specific antibody responses. Collectively, these studies demonstrate a basis by which CD4+ T cells expressing TCRs with low reactivity toward self-peptides can evade negative selection and acquire enhanced autoreactivity following activation by a cross-reactive antigen. PMID- 16702168 TI - When training is insufficient: reflections on capacity development in health promotion in Peru. AB - The international development community has lately recognized the programatic importance of capacity development. Despite growing attention, a consensus is still lacking on appropriate strategies and interventions to strengthen local capacity for development programs. The CHANGE Project designed and implemented a program to contribute to capacity development in health promotion in Peru between 2002 and 2005. This article discusses key lessons learned in the program. Successful capacity development requires the strengthening of institutional networks. Training activities alone are insufficient. Educational opportunities need to be integrated into strategies that aim to institutionalize competencies in specific work contexts and promote the inter-institutional linkages. The experience suggests that capacity development programs need to start by assessing challenges to make competencies effective in job performance. Projects should neither be donor-driven nor depend on individuals. Instead, they should focus on institution building, find creative ways to promote long-term sustainability of capacity development, and require long-term commitment to promote ownership and sustainability. What is needed is an approach that maximizes the impact of programs through collaboration among institutions committed to supporting and absorbing capacity, and to having mechanisms to monitor and change job performance according to the needs of staff, supervisors and other stakeholders. PMID- 16702169 TI - An overview and process evaluation of TeleWalk: a telephone-based counseling intervention to encourage walking in older adults. AB - Despite the benefits associated with a physically active lifestyle, many older adults are insufficiently active to achieve health gain, and also exhibit decreased activity levels with age. Insufficient physical activity in this population is associated with increased morbidity, mortality and demand on health care services and expenditure. There is a clear need for effective intervention to encourage physical activity in older adults. The aim of this paper is to describe the development and participant evaluation of a randomized controlled trial of TeleWalk, a telephone-based motivational counseling intervention to encourage physical activity in adults aged 65 years and older. Participants (N = 186, mean age 74 +/- 6 years) were recruited through their General Practitioner (primary care physician) and randomized to either receive eight telephone counseling sessions and related printed materials over 3 months (intervention group) or participate in outcome assessments only (control group). Intervention group participants were mailed an anonymous evaluation questionnaire on intervention completion. A high response rate was achieved (70%). All respondents (100%) agreed or strongly agreed that a good overall level of service and support was provided, and that the counselor was understanding and supportive. Nearly all respondents agreed or strongly agreed that the service was professional, the counselor advice was helpful and motivating and the information provided was relevant (97, 95 and 89%, respectively). Most (87%) agreed or strongly agreed that the telephone calls encouraged them to be physically active. Among the participants who received printed material, most agreed or strongly agreed that these encouraged them to become or remain active. Findings from this evaluation can be used to inform such interventions and ensure their relevance to community dwelling older adults. PMID- 16702170 TI - Evidence behind the WHO guidelines: hospital care for children: should s/c adrenaline, hydrocortisone or antihistamines be used as premedication for snake antivenom? PMID- 16702171 TI - The cost of cardiovascular disease: rising, declining or staying still? AB - Epidemiological, demographic and health technology factors combined are likely to inflate future healthcare CVD costs, with an older and healthier population consuming higher levels of resources required for treatment. PMID- 16702172 TI - Cost of cardiovascular diseases in the United Kingdom. AB - OBJECTIVE: To estimate the economic burden of cardiovascular disease (CVD) in the United Kingdom, including health and non-healthcare costs, and the proportion of total CVD cost due to coronary heart disease (CHD) and cerebrovascular disease. DESIGN AND SETTING: Prevalence-based approach to assess CVD-related costs from a societal perspective. PATIENTS: All UK residents in 2004 with CVD (International classification of diseases, 10th revision (ICD-10) codes I00-I99) and subgroups with CHD (ICD-10 codes I20-I25) or cerebrovascular disease (ICD-10 codes I60 I69). MAIN OUTCOME MEASURES: Healthcare costs were estimated from expenditure on community health and social services, accident and emergency care, hospital care, rehabilitation and drugs. Non-healthcare costs were estimated from data on informal care and from productivity losses arising from morbidity and premature death. RESULTS: CVD cost the UK economy 29.1 billion pound in 2004, with CHD and cerebrovascular disease accounting for 29% (8.5 billion pound) and 27% (8.0 billion pound) of the total, respectively. The major cost component of CVD was health care, which accounted for 60% of the cost, followed by productivity losses due to mortality and morbidity, accounting for 23%, with the remaining 17% due to informal care-related costs. CONCLUSIONS: CVD is a leading public health problem in the UK measured by the economic burden of disease. This study identified the size and main components of that burden, and will help to inform decisions about research priorities and to monitor the impact of policy initiatives. PMID- 16702174 TI - Endothelial function in HIV-infected patients with low or mild cardiovascular risk. AB - BACKGROUND: Highly active antiretroviral therapy for HIV-infected patients is associated with metabolic side effects, which could cause an increased cardiovascular risk in these patients. Non-invasive study of endothelial function by brachial artery ultrasound can detect subclinical atherosclerosis. Several studies have assessed endothelial function in HIV-infected patients with associated cardiovascular risk factors. OBJECTIVES: The aim of this study is to determine endothelial function in HIV-infected patients under antiretroviral therapy with low or mild coronary risk and lipid levels within the normal range. METHODS: Transversal study including 28 HIV-infected adults (15 receiving antiretroviral therapy and 13 naive) with low or mild cardiovascular risk and 12 healthy controls. Subjects with diabetes mellitus, hypertension, cardiovascular disease, obesity, high cholesterol or high triglyceride levels were excluded. Endothelial function was determined with flow-mediated dilation (FMD) of the brachial artery by ultrasound study. RESULTS: Treated HIV-infected patients had significantly lower FMD (5.93 +/- 3.56) than healthy controls (10.64 +/- 3.08, P = 0.008). Naive patients had an intermediate FMD, but this was not statistically significant. CONCLUSIONS: HIV-infected patients receiving antiretroviral therapy who have low or mild cardiovascular risk and lipid levels within the normal range have endothelial dysfunction compared with healthy controls. PMID- 16702175 TI - Antiproliferative activities of two novel quinuclidine inhibitors against Toxoplasma gondii tachyzoites in vitro. AB - OBJECTIVES: To study the antiproliferative effects of ER119884 and E5700, two quinuclidine-based inhibitors of squalene synthase (SQS), against Toxoplasma gondii tachyzoites in epithelial cells. METHODS: The antiproliferative effects of the quinuclidine derivatives, alone or in combination with epiminolanosterol or antifolates, were analysed, resulting in the construction of isobolograms. The ultrastructure of treated tachyzoites was analysed by transmission electron microscopy. RESULTS: The quinuclidine derivatives demonstrated selective anti-T. gondii activity, arresting parasite growth with IC50 values of 0.66 and 0.23 microM for ER119884 and E5700, respectively, after 24 h of interaction and 0.44 and 0.19 microM after 48 h of interaction. Both compounds induced remarkable alterations in the parasite ultrastructure, such as mitochondrial swelling and the presence of autophagosome-like structures, after 24 h of treatment. Combination of these quinuclidine derivatives with the antifolates sulfadiazine and pyrimethamine produced a synergic effect. When epiminolanosterol was combined with E5700, the effect observed was synergic, whereas the combination with ER119884 produced no interaction. CONCLUSIONS: E5700 and ER119884 demonstrated selective activity against T. gondii tachyzoites and are a possible alternative to be used in association with the current therapy. The ultrastructural alterations observed suggest a possible interference with lipid metabolism. PMID- 16702176 TI - Understanding the role parents play in tennis success: a national survey of junior tennis coaches. AB - OBJECTIVES: To assess coaches' perceptions about the role of parents and their positive and negative behaviours in junior tennis. METHODS: A national survey of 132 United States junior tennis coaches was completed. The extent and seriousness/impact of parent-child interaction problems and positive behaviours were rated. RESULTS: Parents were perceived as very important for junior tennis success. Most parents (59%) that these coaches had worked with were seen as having a positive influence on their player's development. However, the respondents also felt that 36% of parents negatively influenced their child's development. Positive parental behaviours included providing logistical, financial, and social-emotional support, as well as tennis opportunities and unconditional love. Negative parent behaviours included overemphasising winning, holding unrealistic expectations, and criticising their child. CONCLUSIONS: Findings are discussed relative to current sport parenting and athletic talent development research and theorising. The need to educate parents is emphasised. PMID- 16702177 TI - Ultrastructural transformation of gastric parietal cells reverting from the active to the resting state of acid secretion revealed in isolated rat gastric mucosa model processed by high-pressure freezing. AB - To elucidate a functional transformation of gastric parietal cells, we have newly developed an isolated rat gastric mucosa model whose parietal cells exhibited a reverting process from the active to the resting state of acid secretion. Briefly, the parietal cells were treated with cimetidine following prior stimulation of acid secretion in the model, and cryofixed by plunge freezing for light microscopy or high-pressure freezing for electron microscopy. As a result, immunohistochemistry of H(+)/K(+)-ATPase demonstrated a progressive translocation of H(+)/K(+)-ATPase from the apical to the cytoplasmic region. The ultrastructure of parietal cells at 5 min in the reverting phase was quite similar to that of maximally stimulated one. However, the apical microvilli of intracellular canaliculi (IC) changed bulbous by degrees, resulted in complete occlusion of IC at 60 min in the reverting phase. The apical membranes were subsequently internalized into the cytoplasm forming unique penta-laminar membranes. Interestingly, at 90 min in the reverting phase, the penta-laminar membranes formed a number of multilamellar autophagosomes that were intensely labeled for H(+)/K(+)-ATPase. Then, the parietal cells exhibited well-developed Golgi apparatus and lysosomal compartments involving the multilamellar membranes at 105 min, and mostly reverted to their resting conformation at 120 min in the reverting phase. Corresponding to the ultrastructural changes of microvilli, the immunohistochemistry of ezrin showed a dissociation of ezrin from the apical region at 30 min in the reverting phase. The present findings provide new insights into the functional transformation in gastric parietal cells reverting to their resting conformation. PMID- 16702178 TI - Mantle cell lymphoma: an update on management. AB - Although response rates are increased, the addition of rituximab to induction chemotherapy has not yet been proven to extend the progression-free and overall survival benefits of chemotherapy alone. In first remission, high-dose therapy plus stem cell rescue improves time to treatment failure and progression-free survival when compared with maintenance interferon alpha. However, relapse rate does not reach a plateau. Radioimmunotherapy has substantial single-agent activity and when combined with chemotherapy may provide a platform onto which rituximab or autologous stem cell transplantation can be added. Targeted therapies are also showing promise and may have a role in maintenance and/or initial therapy. PMID- 16702179 TI - Extreme Hodgkin's lymphoma: current problem areas. AB - In Hodgkin's lymphoma therapy, there remain areas of extreme difficulty. This article briefly explores these areas and provides evidence that risk-related therapy and web-based data collection programmes can facilitate progress. Unpublished data from the northern region of the UK suggest that risk-based therapy among poor-prognosis Hodgkin's lymphoma patients aged 15-19 years is improving outcome. An index developed at the Memorial Sloan-Kettering Cancer Centre (MSKCC) for patients undergoing salvage therapy shows that new second-line regimens are urgently needed for those with poor prognosis. In primary therapy and relapse, both adolescent and adult patients with Hodgkin's lymphoma should receive treatment tailored to their degree of risk. Elderly patients with Hodgkin's lymphoma are also difficult to treat and fewer than 2% enter randomised clinical trials. We require new means of encouraging international pooling of data and treatment evaluation in such patients. The SHIELD project (Study of Hodgkin's lymphoma in the Elderly Lymphoma Database; www.shieldstudy.co.uk) is now successfully providing on-line registration in the ongoing phase II study of VEPEMB (vinblastine, cyclophosphamide, procarbazine, prednisolone, mitoxantrone and bleomycin) in the elderly and guidance on obtaining ethical approval for participation. PMID- 16702180 TI - Gemcitabine, ifosfamide, oxaliplatin and rituximab (R-GIFOX), a new effective cytoreductive/mobilizing salvage regimen for relapsed and refractory aggressive non-Hodgkin's lymphoma: results of a pilot study. AB - BACKGROUND: The prognosis of patients with aggressive non-Hodgkin's lymphoma (NHL) relapsing or progressing after front-line therapy remains poor. Since high dose therapy (HDT) with autologous stem cell transplantation (ASCT) can cure a proportion of such patients, provided that a substantial tumor shrinkage is achieved, the development of more effective and less toxic salvage regimens remains a major challenge. We evaluated the clinical activity, toxicity and mobilizing capacity of a new salvage regimen, which combines gemcitabine and oxaliplatin with ifosfamide and rituximab (R-GIFOX) in patients with relapsed and refractory CD20(+) NHL. PATIENTS AND METHODS: Patients were scheduled to receive three courses of therapy followed by mobilization and ASCT or three more courses if ineligible for ASCT. R-GIFOX consisted of rituximab (375 mg/m(2) on day 1), gemcitabine (1000 mg/m(2) on day 2), oxaliplatin (130 mg/m(2) on day 3) and ifosfamide (5 g/m(2) on day 3) as a 24-h single infusion in patients aged < or =65 years, or fractionated over 3 days (days 3-5) in patients aged >65 years. Treatment was given every 2 weeks with G-CSF support (5 microg/kg/day or 10 microg/kg/day at the end of the third course for stem cell mobilization). Responses were evaluated by the integrated FDG-PET/IWC criteria after the third course and at the end of the entire program. RESULTS: Fourteen patients (median age 63 years, range 37-78 years) with relapsed (n = 9) or primary progressive (n = 5) aggressive (diffuse large cell, mantle cell, follicular G3), advanced (stage IV 71%), poor risk (IPI 3-5 50%) NHL were accrued in this pilot study. Patients had received a median of two previous treatment lines (range 1-4). The median number of R-GIFOX courses delivered was 4 (range 1-6). Thirteen patients completed at least three courses of therapy and were evaluable for response. The overall response rate assessed after three courses of R-GIFOX was 77%, with seven complete responses and three partial responses. Effective CD34(+) cell mobilization was obtained in four of six eligible patients and two had ASCT. Hematologic and extra-hematologic toxicity was tolerable. Failure-free survival was 79.6% at median follow-up of 6 months (range 2-12). Molecular remissions were documented in two patients with mantle cell NHL. CONCLUSIONS: Based on the results of this pilot study, we conclude that the R-GIFOX regimen is feasible, tolerable, effective and able to mobilize peripheral stem cells in patients with relapsed and refractory aggressive NHL. PMID- 16702181 TI - Outpatient fractionated ifosfamide, carboplatin and etoposide as salvage therapy in relapsed and refractory non-Hodgkin's and Hodgkin's lymphoma. AB - We have treated 75 transplant-eligible patients with relapsed or refractory lymphoma using an outpatient-based fractionated regimen of ifosfamide, carboplatin and etoposide (ICE) for both salvage and stem cell mobilisation. Patients included DLBC (n = 33), follicular (n = 23), NK/T-cell (n = 3), mantle cell (n = 3) and Hodgkin's lymphoma (n = 13). Cycles of outpatient ICE were given every 21 days and consisted of: ifosfamide 5000 mg/m(2) i.v. fractionated into three equally divided doses and infused over 2-3 h on days 1-3, carboplatin (mg dose = 5 x AUC) i.v. over 1 h on day 1; and etoposide 100 mg/m(2) i.v. daily on days 1-3, plus filgrastim 5 microg/kg/day. Most patients with indolent lymphoma also received rituximab. The median age of patients was 52 years (range 26-69 years). Patients received a mean of 2.8 cycles of ICE. Non-haematological toxicities included grade 1/2 CNS toxicity in four patients, cardiac toxicity in two, reversible renal impairment and haematuria in one each. Haematological toxicity included grades III/IV thrombocytopenia and neutropenia with at least one cycle of ICE in 71% and 72% of patients, respectively. The median time to PBSC harvest was 14 days (range 10-20 days), while the median CD34(+) cell yield was 4.8 x 10(6)/kg (range 2.3-37.8). Five patients (7%) failed to mobilise PBSCs. The overall response rate to ICE was 89%, comprising 29% who achieved a CR and 60% who achieved a PR; for DLBCL, the overall response rate was 85% including 36% who achieved a CR and 49% who exhibited a PR. At a median follow-up of 24 months, the Kaplan-Meier estimates of the overall and event-free survival for all patients were 65% and 42%, respectively. For patients with DLBCL overall and event-free survival figures were 51% and 35%, respectively, at a median follow-up of 14 months. These data confirm the efficacy and tolerability of outpatient fractionated ICE as both a salvage and mobilisation regimen in relapsed/refractory lymphoma. PMID- 16702182 TI - Randomised phase III study of R-ICE versus R-DHAP in relapsed patients with CD20 diffuse large B-cell lymphoma (DLBCL) followed by high-dose therapy and a second randomisation to maintenance treatment with rituximab or not: an update of the CORAL study. AB - The multicentre phase III CORAL study aims to guide choice of salvage chemotherapy in diffuse large B-cell lymphoma (DLBCL) and assess the role of rituximab maintenance after autologous stem cell transplantation (ASCT). Patients are first randomised between ICE (ifosfamide, carboplatin, etoposide) and DHAP (dexamethasone, ara-C and cisplatin), both combined with rituximab (R-ICE or R DHAP). After three courses, responders are treated by ASCT with BEAM. A second randomisation then allocates patients to maintenance treatment with rituximab 375 mg/m(2), one injection every 2 months six times, or observation. Accrual to the study is now proceeding well and the planned 400 patients are likely to be enrolled within the next 1.5 years. Results to date are very preliminary but suggest encouraging rates of response. However, they also indicate that initial exposure to rituximab may increase the difficulty of salvaging patients who fail first-line therapy. PMID- 16702183 TI - Ifosfamide: pharmacokinetic properties for central nervous system metastasis prevention. AB - The incidence of central nervous system (CNS) recurrence in patients with lymphoma is about 5%. Nevertheless, this complication is very serious because it is almost always fatal. Its incidence is not sufficiently high to warrant the use of CNS prophylaxis in all patients. The identification of subgroups for whom CNS prophylaxis may be of benefit is therefore important and the age-adjusted international prognostic index (aa-IPI) may be useful in this respect. Ifosfamide (IFO) is a widely used antitumor agent, requiring activation to isophosphoramide mustard (IPM) for DNA alkylation. IFO anabolism occurs through the hepatic microsomal cytochrome P450 system. As with the majority of antineoplastic agents, IFO has toxic side-effects. These include neurotoxicity due to the chloroacetaldehyde (CAA) catabolite. However, the incidence of neurotoxicity is low when IFO is administered as a continuous intravenous infusion. Both inactive IFO and active IPM cross the blood-brain barrier, making IFO treatment effective in the prevention of CNS metastasis in lymphoma patients at high risk of recurrence. The benefit/risk ratio for such patients should evaluated. PMID- 16702184 TI - Pretreatment prognostic factors and outcome in patients with relapsed or primary refractory diffuse large B-cell lymphoma treated with second-line chemotherapy and autologous stem cell transplantation. AB - The age-adjusted International Prognostic Index assessed before salvage therapy with ICE (ifosfamide, carboplatin, etoposide) predicts outcome in patients with relapsed or primary refractory diffuse large B-cell lymphoma (DLBCL). Patients can be stratified according to this index into favorable and unfavorable cohorts. Subsequently we attempted to determine if the cell of origin as determined by immunohistochemistry would predict outcome, as it had in the first-line setting. However, none of the molecular markers, which are prognostic in first-line therapy, nor immunohistochemical classification by cell of origin, relate to survival outcome of DLBCL patients in the second-line setting, implying that dose intensification of therapy can overcome the prognostic import of these unfavourable risk factors. PMID- 16702185 TI - Histopathology in the light of molecular profiling. AB - The 2001 WHO classification distinguishes five variants (centroblastic, immunoblastic, plasmablastic, anaplastic and T-cell rich) and three subtypes (primary mediastinal, intravascular and primary effusion large B-cell lymphoma) of diffuse large B-cell lymphomas (DLBCLs).The recognition of the three subtypes as distinct disease entities can be considered as an advance in our understanding of these tumours. However, the variants of DLBCLs, which significantly outnumber the subtypes in frequency, represent an unresolved area. Gene expression profiling (GEP) of the variants led to a discrepancy in results and produced more questions than answers. The authors, therefore, initiated a multi-institutional collaborative research project in Germany aimed at a subtle morphologic, genomic and transcriptional characterisation of DLBCLs and Burkitt lymphoma (BL). We included BL in our study for two reasons: (1) it belongs to aggressive B-NHLs; and (2) at present, there are no reliable criteria that can be applied to distinguish BL from DLBCL. The GEPs derived from 200 patient samples were correlated with reviewed histology, the degree of genetic imbalances and clinical features. The results of this approach show that: (i) the DLBCL can be divided into more than four molecular groups; and (ii) the BL cases, identified by the consensus of five out of six lymphoma expert pathologists, displayed a genomic and gene expression profile that was clearly distinct from those of most DLBCLs. The group of DLBCLs that resembled BL in their GEP had a remarkably good prognosis, whereas those that differed in their GEP from the consensus BLs had unfavourable survival rates. In conclusion, combined application of genomic and gene expression profiling in conjunction with consensus reviewed histology and clinical features, appears to be a reliable approach that enables a reproducible and clinically meaningful characterisation of mature aggressive B-NHLs. PMID- 16702186 TI - Managing large cell lymphoma. AB - Rituximab has greatly improved standard cyclophosphamide, doxorubicin, vincristine and prednisone (CHOP) chemotherapy, but prospects remain poor in patients with adverse risk factors. We need to know whether the addition of rituximab also improves the efficacy of more intensive treatments and how to use molecular profiling to guide us in introducing other novel agents. PMID- 16702187 TI - Life-threatening malfunction of implantable cardiac devices. PMID- 16702188 TI - The impact of standalone call centres and GP cooperatives on access to after hours GP care: a before and after study adjusted for secular trend. AB - BACKGROUND: The After Hours Primary Medical Trials were initiated by the Australian government to redress difficulties in after hours (AH) GP care in areas of high need. The study's objective is to study the impact of two standalone call centres and one GP cooperative offering comprehensive services, in improving consumer access to services for residents of a defined geographic area. METHODS: A pre-post design was used to evaluate their impact after adjusting for secular trend at a national level. Access was considered in terms of availability, accessibility, affordability, acceptability and responsiveness of care. Unmet need and ease of obtaining AH telephone professional medical advice were also considered. Pre-trial and post-trial telephone surveys of two separate random samples of approximately 350 households using AH services in each trial area as well as in a national sample outside the trial areas. RESULTS: Consumer acceptability and affordability increased in residents in the area served by the GP cooperative. Access, however measured, did not improve in either of the standalone call centre areas. Reduction in unmet need approached but did not achieve statistical significance in most but not all trial areas. CONCLUSIONS: Improvements in access in the GP cooperative conformed to expectations based on current and pre-existing AH care arrangements put in place. Absence of improvements in access in the standalone call centres did not conform to expectations but may be partly explained by the reductions in consumer acceptability, following introduction of telephone triage systems reported elsewhere. PMID- 16702189 TI - Genomewide scans in North American families reveal genetic linkage of essential tremor to a region on chromosome 6p23. AB - Essential tremor (ET) is the most prevalent adult-onset movement disorder showing evidence of non-random accumulation in some families. ET has previously been mapped to genetic loci on chromosomes 2p and 3q, but no causative genes identified. We conducted genomewide linkage screening with subsequent fine mapping in seven large North American families comprising a total of 325 genotyped individuals that included 65 patients diagnosed as definite ET. Linkage analysis was based on methodology implemented in SimWalk2 and LINKAGE programs. A multigenerational family revealed suggestive linkage to a locus on chromosome 6p23 with maximal nonparametric linkage (NPL) multipoint score 3.281 (P = 0.0005) and parametric multipoint log of the odds (LOD) score 2.983. A second family showed positive linkage to the same 6p23 region with a maximal NPL score 2.125 (P = 0.0075) and LOD score 1.265. Haplotype analysis led to the identification of a 600 kb interval shared by both families. Sequencing of coding regions of 15 genes located in the linked region detected numerous sequence variants, some of them predicting a change of the encoded amino acid, but each was also found in controls. Our findings provide evidence for linkage to a novel susceptibility locus on chromosome 6p23. Analysis of additional ET-affected families is needed to confirm linkage and identify the underlying gene. PMID- 16702190 TI - Impairment of mitochondrial anti-oxidant defence in SOD1-related motor neuron injury and amelioration by ebselen. AB - There is now compelling evidence of mitochondrial dysfunction in motor neuron disease (MND), but the molecular basis of these abnormalities is unknown. It is also unclear whether the observed mitochondrial dysfunction plays a central role in disease pathogenesis, and if so, whether its amelioration might present therapeutic opportunities. We adopted a candidate generation approach using proteomics to screen for changes in mitochondrial protein expression in a well validated cell-culture model of superoxide dismutase 1 (SOD1) related familial MND (fMND). Changed proteins were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectroscopy. Protein candidates included apoptotic regulators, anti-oxidants and components of the electron transport chain. Confirmatory Western blotting was performed, and validated protein expression changes were further investigated. Peroxiredoxin 3 (Prx3), a mitochondrial thioredoxin-dependent hydroperoxidase, is downregulated in the presence of mutant SOD1 in both our cell-culture model and in the spinal cord mitochondria of mutant SOD1 transgenic mice. We confirm the expression of Prx3 within the mitochondria of spinal motor neurons in mouse and humans by immunohistochemistry. Using quantitative real-time PCR (Q-PCR), we show that Prx3 is also downregulated in spinal motor neurons from patients with both sporadic (sMND) and SOD1-related fMND. In a disease characterized by oxidative stress, this represents a potentially important deficit in mitochondrial anti-oxidant defence. Recent evidence suggests that oxidative stress from aberrant copper chemistry may not play a major part in the pathogenesis of SOD1-related fMND. From the results of this study we propose disruption of mitochondrial anti oxidant defence as an alternative mechanism whereby mutant SOD1 may generate oxidative stress within motor neurons. We further demonstrate that ebselen, an anti-oxidant drug already safely used in human studies and that acts as a Prx mimic, is able to ameliorate the toxicity of mutant SOD1 in our cell-culture model. We conclude by showing that ebselen is capable of inducing transcription of the anti-oxidant response element (ARE) and postulate that ebselen may act both by the transcriptional upregulation of anti-oxidant proteins, and directly as an anti-oxidant in its own right. PMID- 16702191 TI - PINK1 protein in normal human brain and Parkinson's disease. AB - Parkinson's disease is a common incurable neurodegenerative disease whose molecular aetiology remains unclear. The identification of Mendelian genes causing rare familial forms of Parkinson's disease has revealed novel proteins and pathways that are likely to be relevant in the pathogenesis of sporadic Parkinson's disease. Recently, mutations in a novel gene, PINK1, encoding a 581 amino acid protein with both mitochondrial targeting and serine/threonine kinase domains, were identified as a cause of autosomal recessive parkinsonism. This provided important evidence for the role of the mitochondrial dysfunction and kinase pathways in neurodegeneration. In this study, we report the first characterization of the PINK1 protein in normal human and sporadic Parkinson's brains, in addition to Parkinson's cases with heterozygous PINK1 mutations. The possible role of the PINK1 protein was also assessed in a number of neurodegenerative diseases characterized by proteinaceous inclusions. For these studies, rabbit polyclonal antibodies were raised against two peptide sequences within the N-terminal hydrophilic loops of PINK1 protein. Using immunohistochemistry and western blotting we were able to demonstrate that PINK1 is a ubiquitous protein expressed throughout the human brain and it is found in all cell types showing a punctate cytoplasmic staining pattern consistent with mitochondrial localization. Fractionation studies of human and rat brain confirm that PINK1 is localized to the mitochondrial membranes. In addition, we show that PINK1 is detected in a proportion of Lewy bodies in cases of sporadic Parkinson's disease and Parkinson's disease associated with heterozygous mutations in the PINK1 gene, which are clinically and pathologically indistinguishable from the sporadic cases. PINK1 was absent in cortical Lewy bodies, in neurofibrillary tangles in Alzheimer's disease, progressive supranuclear palsy and corticobasal degeneration, and in the glial and neuronal alpha-synuclein positive inclusions in multiple system atrophy. These studies provide for the first time in vivo morphological and biochemical evidence to support a mitochondrial localization of PINK1 and underpin the significance of mitochondrial dysfunction in the pathogenesis of nigral cell degeneration in Parkinson's disease. PMID- 16702193 TI - Does socio-economic status and health consciousness influence how women respond to health related messages in media? AB - During the past few decades, people have been increasingly exposed to health related messages in the mass media, conveying recommendations for healthy lifestyles. The present study investigates whether these messages represent a stressor, and whether coping responses increase levels of motivation or levels of negative affect. A sample of 403 women aged 45 years were surveyed twice, at an interval of 4 weeks. A substantial proportion of the participants perceived the health messages to be stressful (increased levels of threat). Overall, the participants reported a greater use of adaptive than non-adaptive coping when exposed to the health messages. Socio-economic status (defined in educational terms) was negatively correlated with non-adaptive coping, while health consciousness was positively correlated with adaptive coping. Adaptive coping was positively related, and non-adaptive coping was negatively related, to intentions and behaviours. Non-adaptive coping was associated with stronger negative emotions. The results indicate that less-educated women tend to respond more non adaptively to health messages than more-educated women; for the former group, this has negative consequences in terms of increased levels of negative emotions and decreased levels of motivation to engage in healthy behaviours. PMID- 16702194 TI - Website physical activity interventions: preferences of potential users. AB - Information and communication technologies (particularly websites and e-mail) have the potential to deliver health behavior change programs to large numbers of adults at low cost. Controlled trials using these new media to promote physical activity have produced mixed results. User-centered development methods can assist in understanding the preferences of potential participants for website functions and content, and may lead to more effective programs. Eight focus group discussions were conducted with 40 adults after they had accessed a previously trialed physical activity website. The discussions were audio taped, transcribed and interpreted using a themed analysis method. Four key themes emerged: structure, interactivity, environmental context and content. Preferences were expressed for websites that include simple interactive features, together with information on local community activity opportunities. Particular suggestions included online community notice boards, personalized progress charts, e-mail access to expert advice and access to information on specific local physical activity facilities and services. Website physical activity interventions could usefully include personally relevant interactive and environmentally focused features and services identified through a user-centered development process. PMID- 16702192 TI - Non-invasive mapping of corticofugal fibres from multiple motor areas--relevance to stroke recovery. AB - Recovery of motor function after subcortical stroke appears to be related to the integrity of descending connections from the ipsilesional cortical motor system, a view supported by the observation of greater than normal movement-related activation in ipsilesional motor regions in chronic subcortical stroke patients. This suggests that damage to the descending output fibres from one region of the cortical motor system may be compensated by activity in areas that retain corticofugal outputs. Though the trajectories of corticofugal fibres from each major component of the motor system through the corona radiata and internal capsule are well described in non-human primates, they have not been described fully in humans. Our study set out to map the trajectories of these connections in a group of healthy volunteers (8 male, 4 female; age range = 31-68 years, median = 48.5 years) and establish whether this knowledge can be used to assess stroke-induced disconnection of the cortical motor system and better interpret functional reorganization of the cortical motor system. We describe the trajectories of the connections from each major component of the motor system to the cerebral peduncle using diffusion-weighted imaging and probabilistic tractography in normal subjects. We observed good reproducibility of these connections over subjects. The comparative topography of these connections revealed many similarities between humans and other primates. We then inferred damage to corticofugal pathways in stroke patients (n = 3) by comparing the overlap between regions of subcortical white matter damage with the trajectories of the connections to each motor area. In a small series of case studies, we found that inferred disconnections could explain enhanced hand-grip-related responses, as assessed with functional MRI, in the ipsilesional motor system. These results confirm that selective disruption of motor corticofugal fibres influences functional reorganization and outcome in individual patients. PMID- 16702195 TI - Physical activity promotion in general practices of Barcelona: a case study. AB - This case study aimed to generate explanations for the lack of integration of physical activity (PA) promotion in general practices of Barcelona, the capital of Catalonia. This explanatory study adopted a qualitative approach, based on three techniques; focus groups (n = 3), semi-structured (n = 25) and short individual interviews (n = 5). These approaches explored the wider environment surrounding primary care from a range of distinctive professional and personal perspectives. Participants were recruited as patients (n = 20), policy makers (n = 6), academics (n = 5), PA professionals (n = 3), medical doctors (n = 3), researchers (n = 2), media employees (n = 2) and one social worker. Phenomenological techniques were used for data coding and interpretation. Contributors confirmed the final interpretation. Three main factors underpinned the lack of integration of PA promotion approaches. PA promotion delivery rarely accounted for either patients' individual needs or the circumstances that influenced their interest in PA promotion. This was a missed opportunity in promotional consultations. There was also a lack of official support for general practitioner-based PA promotion. Finally, primary care staff intentionally isolated their practice from other professionals and/or services in the community. Community-based PA promotion could be better integrated by (i) introducing stage-based strategies, (ii) creating top-down approaches and (iii) connecting primary care with other professionals and institutions in the community. PMID- 16702196 TI - An experimental study of effects on schoolchildren of exposure to point-of-sale cigarette advertising and pack displays. AB - By creating a sense of familiarity with tobacco, cigarette advertising and bold packaging displays in stores where children often visit may help to pre-dispose them to smoking. A total of 605 ninth-grade students were randomly allocated to view a photograph of a typical convenience store point-of-sale which had been digitally manipulated to show either cigarette advertising and pack displays, pack displays only or no cigarettes. Students then completed a self-administered questionnaire. Compared with those who viewed the no cigarettes, students either in the display only condition or cigarette advertising condition perceived it would be easier to purchase tobacco from these stores. Those who saw the cigarette advertising perceived it would be less likely they would be asked for proof of age, and tended to think a greater number of stores would sell cigarettes to them, compared with respondents who saw no tobacco products. Respondents in the display only condition tended to recall displayed cigarette brands more often than respondents who saw no cigarettes. Cigarette advertising similarly influenced students, and tended to weaken students' resolve not to smoke in future. Retail tobacco advertising as well as cigarette pack displays may have adverse influences on youth, suggesting that tighter tobacco marketing restrictions are needed. PMID- 16702197 TI - Anti-RANKL therapy--implications for the bone-vascular-axis in CKD? Denosumab in post-menopausal women with low bone mineral density. PMID- 16702198 TI - The effect of haemodialysis on transcutaneous oxygen tension in patients with diabetes-a pilot study. AB - BACKGROUND: Established renal failure in diabetes is associated with a high incidence of foot ulcers and gangrene, and these are major causes of morbidity and mortality. It has been suggested that this problem is particularly associated with the onset of renal replacement therapy, and since there is evidence that haemodialysis causes hypoxaemia, we have undertaken a pilot study to determine the effect of haemodialysis on lower limb transcutaneous oxygen tension (TcpO2). METHODS: TcpO2 was measured on the dorsum of the foot through a single dialysis treatment and over the succeeding 4 h using a transcutaneous monitor (TCM400/3, Radiometer Ltd, Copenhagen, Denmark). RESULTS: The median age (range) of the 10 (7 male) participants was 73 (58-83) years. The median duration of diabetes was 16.5 (7-30) years and that of dialysis treatment 29 (10-88) months. The median (range) baseline TcpO2 on the dorsum of the foot was 54.5 (51-77) mmHg and 54.0 (24-87) at the end of dialysis. Median TcpO2 at 1, 2 and 4 h after the end of dialysis was 50.0 (33-81), 49.0 (24-78) and 47.0 (20-78) mmHg. Analysis by ANOVA suggested a trend towards a difference between median TcpO2 concentrations at different time points (F(1.752, 15.765) = 3.359, P = 0.066). CONCLUSIONS: The data identified a trend towards a fall in lower limb TcpO2, and that this fall continued for at least 4 h after the end of treatment. Dialysis-associated lower limb hypoxia may be a factor leading to the increased incidence of critical limb ischaemia in this group and further work is needed to define its cause and implications for clinical care. PMID- 16702199 TI - A patient recently transplanted with a living donor kidney develops severe neurological symptoms. PMID- 16702200 TI - Carbimazole therapy in the setting of end-stage renal disease and haemodialysis. PMID- 16702201 TI - Long-term outcome of idiopathic retroperitoneal fibrosis treated with surgical and/or medical approaches. AB - BACKGROUND: Retroperitoneal fibrosis is a severe disease that affects the ureters, causing renal insufficiency in three-quarters of patients. The optimal treatment is far from being established. METHODS: Seventeen patients with idiopathic retroperitoneal fibrosis and ureteral entrapment followed in our unit for at least 1 year were selected for this study. At presentation 13 patients had renal insufficiency. All patients received steroids, associated with ureterolysis in five (group 1), with azathioprine in six (group 2) and with tamoxifen in six (group 3). Four patients of group 2 and five of group 3 received ureteral stenting or nephrostomy. There were no significant differences among the three groups or the clinical and biochemical characteristics at presentation. RESULTS: All patients of groups 1 and 2 entered remission after therapy. One patient from group 3 did not respond to therapy. During a mean follow-up of 56 +/- 41 months, three patients (two from group 1, one from group 2, 18%) had a recurrence of the disease, which fully responded to retreatment in all three cases. At the last observation, all patients were alive; three patients (18%) had renal insufficiency, of them one from group 1 had to start dialysis 6 years after ureterolysis, one patient from group 2 and one from group 3 had serum creatinine of 1.5 mg/dl. Renal survival was 100% at 5 years and 80% at 10 years. CONCLUSIONS: In most patients, each of the three different therapeutic approaches restored renal function and significantly reduced the fibrotic mass in the short term and maintained stable serum creatinine in the long-term. PMID- 16702202 TI - Is tacrolimus for childhood steroid-dependent nephrotic syndrome better than ciclosporin A? PMID- 16702203 TI - Are prediction equations for glomerular filtration rate useful for the long-term monitoring of type 2 diabetic patients? AB - BACKGROUND: The aim of this study was to compare the accuracy of prediction equations [modification of diet in renal disease (MDRD), simplified MDRD, Cockcroft-Gault (CG), reciprocal of creatinine and creatinine clearance] in a cohort of patients with type 2 diabetes. METHODS: A total of 525 glomerular filtration rates (GFRs) using (125)I-iothalamate were carried out over 10 years in 87 type 2 diabetic patients. Accuracy was evaluated at three levels of renal function according to the baseline values obtained with the isotopic method: hyperfiltration (GFR: >140 ml/min/1.73 m(2); 140 isotopic determinations in 27 patients), normal renal function (GFR: 140-90 ml/min/1.73 m(2); 294 isotopic determinations in 47 patients) and chronic kidney disease (CKD) stages 2-3 (GFR: 30-89 ml/min/1.73 m(2); 87 isotopic determinations in 13 patients). The annual slope for GFR (change in GFR expressed as ml/min/year) was considered to ascertain the variability in the equations compared with the isotopic method during follow-up. Student's t-test was used to determine the existence of significant differences between prediction equations and the isotopic method (P < 0.05 with Bonferroni adjusted for five contrast tests). RESULTS: In the subgroup of patients with hyperfiltration, a GFR slope calculated with (125)I-iothalamate 4.8 +/- 4.7 ml/min/year was obtained. GFR slope in patients with normal renal function was -3.0 +/- 2.3 ml/min/year. In both situations, all equations presented a significant underestimation compared with the isotopic GFR (P < 0.01; P < 0.05). In the subgroup of CKD stages 2-3, the slope for GFR with (125)I iothalamate was -1.4 +/- 1.8 ml/min/year. The best prediction equation compared with the isotopic method proved to be MDRD with a slope for GFR of -1.4 +/- 1.3 ml/min/year (P: NS) compared with the CG formula -1.0 +/- 0.9 ml/min/year (P: NS). Creatinine clearance presented the greatest variability in estimation (P < 0.001). CONCLUSIONS: In the normal renal function and hyperfiltration groups, none of the prediction equations demonstrated acceptable accuracy owing to excessive underestimation of renal function. In CKD stages 2-3, with mean serum creatinine > or =133 micromol/l (1.5 mg/dl), the MDRD equation can be used to estimate GFR during the monitoring and follow-up of patients with type 2 diabetes receiving insulin, anti-diabetic drugs or both. PMID- 16702204 TI - The influence of socio-demographic factors, treatment perceptions and attitudes to living donation on willingness to consider living kidney donor among kidney transplant candidates. AB - BACKGROUND: Living donor, compared with cadaveric donor kidney transplantation, is associated with prolonged graft survival and increased life expectancy for patients with end-stage renal disease (ESRD). However, access to living donor transplantation remains variable. Our objective was to further our understanding of the factors associated with the willingness to consider living donor kidney transplant (LDKTx). METHODS: All patients from two tertiary care hospitals being assessed for a LDKTx (n = 81), and a random sample of patients on the waiting list for a cadaveric kidney transplant (CDKTx, n = 133) were invited to participate. A mail survey assessed participants' socio-demographic characteristics, and perceptions of dialysis, CDKTx and LDKTx. Multivariable logistic regression was used to assess the influence of these factors on willingness to consider LDKTx. RESULTS: Adjusting for patient age, employment status and number of close relatives, willingness to consider an LDKTx was independently associated with a lower perceived risk of peri-operative complications to the donor (adjusted OR 3.59, P = 0.02), the perception that the recipient would live longer following living donor transplantation (adjusted OR 4.34, P = 0.003) and greater perceived appropriateness of asking a family member to donate a kidney (adjusted OR 5.06, P = 0.007). CONCLUSION: Among the ESRD patients studied, adjusting for key factors that may be associated with access to LDKTx, perceptions about risks to the donor and benefits to the recipient were independently associated with a willingness to consider LDKTx as a treatment option. Provision of standardized comprehensible information about all treatment options to patients with ESRD may improve acceptance rates for living donor transplantation. PMID- 16702206 TI - Open and laparoscopic living donor nephrectomy in Switzerland: a retrospective assessment of clinical outcomes and the motivation to donate. AB - BACKGROUND: Laparoscopic living kidney nephrectomy is thought to be associated with reduced morbidity, when compared to open nephrectomy. The purpose of this study was to explore the impact of these techniques on donors' clinical outcomes, satisfaction and motivation to donate. METHODS: Clinical outcomes were retrospectively compared in 152 open (n = 71) or laparoscopic (n = 81) donor procedures. Donor satisfaction and motivation were assessed with a self administered questionnaire. RESULTS: The complication rate was the same with both procedures and the majority of complications were mild. Laparoscopy was significantly less painful and resulted in an insignificantly faster return to active life. More than 80% of the donors volunteered to donate without pressure. Worries about future health status, pain or scars were not important in the decision to donate. Similarly, only 15% considered the surgical procedure as instrumental for their decision. Few donors currently worried about their health with one kidney and more than 95% of the donors in both groups stated that they would give their kidney again. CONCLUSIONS: Living donor nephrectomy is safe, regardless of the procedure used. Although the laparoscopic nephrectomy offers clear short-term benefits over the open nephrectomy, donors' satisfaction was excellent with both surgical approaches. Moreover, the type of procedure did not seem to influence their decision to donate. PMID- 16702205 TI - Low birth weight does not increase the risk of nephropathy in Finnish type 1 diabetic patients. AB - BACKGROUND: Low birth weight (LBW) has been linked to renal disease both in animal models and human studies. However, the role of birth weight in the development of diabetic nephropathy is unclear. We, therefore, studied the impact of birth weight on the development of diabetic nephropathy and other related traits, such as diabetic retinopathy and macrovascular disease, in Caucasian type 1 diabetic patients. METHODS: Data on size at birth were obtained from original birth certificates in 1543 Finnish patients with type 1 diabetes. The patients were divided into those with low (LBW; below the 10th percentile), normal (NBW; 11-90th percentile) and high birth weight (HBW; above the 90th percentile). RESULTS: Diabetic nephropathy was equally common in the groups with various birth weight (LBW vs NBW vs HBW: 21 vs 20 vs 17%, P = NS). End-stage renal disease (3 vs 5 vs 4%, P = NS), laser-treated retinopathy (31 vs 31 vs 31%, P = NS) and macrovascular disease (5 vs 5 vs 8%, P = NS) were equally prevalent in the various birth weight groups. The time from the onset of diabetes to the onset of diabetic nephropathy was similar irrespective of birth weight (log-rank test; P = NS). CONCLUSIONS: Based on our cross-sectional data, LBW does not have an impact on the development of diabetic nephropathy, laser-treated retinopathy or macrovascular disease later in life in Caucasians with type 1 diabetes. PMID- 16702207 TI - Haematopoietic stem cells--role of calcium-sensing receptor in bone marrow homing. PMID- 16702208 TI - Lithium effectively complements vasopressin V2 receptor antagonist in the treatment of hyponatraemia of SIADH rats. AB - BACKGROUND: Although, pharmacological intervention with a selective arginine vasopressin (AVP) V(2) receptor antagonist has been demonstrated to be effective for syndrome of inappropriate secretion of antidiuretic hormone (SIADH), its long term administration has some therapeutic limitations. Lithium, a drug for bipolar disorders, has been known to cause nephrogenic diabetes insipidus by reducing kidney-specific apical water channel, aquaporin 2 (AQP2) expression in the collecting ducts. However, its pharmacological efficacy for SIADH still remains to be elucidated. METHODS: Hyponatraemia was induced in male Sprague-Dawley rats by water loading and subcutaneous infusion of 1-deamino-8-D-arginine vasopressin. For the treatment, lithium chloride (LiCl) was administered singly or in combination with OPC-31260 and/or furosemide for 7 days. Protein expression of AQP2 was examined by western blotting at the end of the observation period. RESULTS: The LiCl administration elevated serum sodium levels in a dose-dependent manner. The therapeutic effect started 3 days after the initial administration and gradually increased. Western blot analysis at the end of the treatment demonstrated dose-dependent reduction of AQP2 protein expression. Additional administration of LiCl (100 mg/kg/day, the dose demonstrated to maintain serum lithium concentration within therapeutic range) to low dose OPC-31260 maintained well the initial elevation of serum sodium level during the treatment. Western blot analysis after combination therapy demonstrated the absence of re-increase in AQP2 expression noted at the end of OPC-31260 treatment. However, further additive effect could not be obtained even when both LiCl and furosemide were added together to low dose OPC-31260. CONCLUSIONS: Although the single effect of therapeutic dose of lithium was weak, it effectively and safely compensated for the therapeutic limitations of a low dose of AVP V(2) receptor antagonist for SIADH by reducing AQP2 expression. PMID- 16702209 TI - The early-immediate gene EGR-1 is induced by transforming growth factor-beta and mediates stimulation of collagen gene expression. AB - Transforming growth factor-beta (TGF-beta) stimulates collagen synthesis and accumulation, and aberrant TGF-beta signaling is implicated in pathological organ fibrosis. Regulation of type I procollagen gene (COL1A2) transcription by TGF beta involves the canonical Smad signaling pathway as well as additional protein and lipid kinases, coactivators, and DNA-binding transcription factors that constitute alternate non-Smad pathways. By using Affymetrix microarrays to detect cellular genes whose expression is regulated by Smad3, we identified early growth response factor-1 (EGR-1) as a novel Smad3-inducible gene. Previous studies implicated Egr-1 in cell growth, differentiation, and survival. We found that TGF beta induced rapid and transient accumulation of Egr-1 protein and mRNA in human skin fibroblasts. In transient transfection assays, TGF-beta stimulated the activity of the Egr-1 gene promoter, as well as that of a minimal Egr-1 responsive reporter construct. Furthermore, TGF-beta enhanced endogenous Egr-1 interaction with a consensus Egr-1-binding site element and with GC-rich DNA sequences of the human COL1A2 promoter in vitro and in vivo. Forced expression of Egr-1 by itself caused dose-dependent up-regulation of COL1A2 promoter activity and further enhanced the stimulation induced by TGF-beta. In contrast, the TGF beta response was abrogated when the Egr-1-binding sites of the COL1A2 promoter were mutated or deleted. Furthermore, Egr-1-deficient embryonic mouse fibroblasts showed attenuated TGF-beta responses despite intact Smad activation, and forced expression of ectopic EGR-1 in these cells could restore COL1A2 stimulation in a dose-dependent manner. Taken together, these findings identify Egr-1 as a novel intracellular TGF-beta target that is necessary for maximal stimulation of collagen gene expression in fibroblasts. The results therefore implicate Egr-1 in the profibrotic responses elicited by TGF-beta and suggest that Egr-1 may play a new and important role in the pathogenesis of fibrosis. PMID- 16702210 TI - Zinc finger protein Wiz links G9a/GLP histone methyltransferases to the co repressor molecule CtBP. AB - G9a is a SET-domain mammalian histone methyltransferase responsible for mono- and dimethylation of lysine 9 in histone H3 (H3K9) at euchromatic regions. Recently we reported that G9a forms a stoichiometric heteromeric complex with another SET domain-containing molecule, GLP/Eu-HMTase1. Although G9a and GLP can independently methylate H3K9 in vitro, G9a/GLP heteromeric formation seems to be essential for their function as a euchromatic H3K9 methyltransferase in vivo. To further elucidate how G9a/GLP-mediated histone methylation and transcriptional regulation are controlled, we purified and characterized G9a complexes from mouse embryonic stem cells. We identified a novel G9a/GLP-associating zinc finger molecule named Wiz that can interact with G9a and GLP independently but is more stable in the G9a/GLP heteromeric complexes. Interestingly, Wiz small inhibitory RNA knocks down not only Wiz but also G9a. GLP deficiency also decreases G9a levels, suggesting that the Wiz/G9a/GLP tri-complex may protect G9a from degradation and that Wiz plays a major role in G9a/GLP heterodimer formation. Furthermore, amino acid sequence analysis of Wiz predicted two potential CtBP binding sites, and indeed CtBP binding to Wiz and association of CtBP with the Wiz/G9a/GLP complex was observed. These data indicate that Wiz not only contributes to the stability of G9a but also links the G9a/GLP heteromeric complex to the CtBP co-repressor machinery. PMID- 16702212 TI - Structural basis for degenerate recognition of natural HIV peptide variants by cytotoxic lymphocytes. AB - It is well established that even small changes in amino acid side chains of antigenic peptide bound to major histocompatibility complex (MHC) protein may completely abrogate recognition of the peptide-MHC (pMHC) complex by the T cell receptor (TCR). Often, however, several nonconservative substitutions in the peptide antigen are accommodated and do not impair its recognition by TCR. For example, a preponderance of natural sequence variants of the human immunodeficiency virus p17 Gag-derived peptide SLYNTVATL (SL9) are recognized by cytotoxic T lymphocytes, which implies that interactions with SL9 variants are degenerate both with respect to the class I MHC molecule and with respect to TCR. Here we study the molecular basis for this degenerate recognition of SL9 variants. We show that several SL9 variants bind comparably well to soluble HLA A2 and to a particular soluble TCR and that these variants are active in the cognate cytotoxicity assay. Natural SL9 variation is restricted by its context in the HIV p17 matrix protein. High resolution crystal structures of seven selected SL9 variants bound to HLA-A2 all have remarkably similar peptide conformations and side-chain dispositions outside sites of substitution. This preservation of the peptide conformation despite epitope variations suggests a mechanism for the observed degeneracy in pMHC recognition by TCR and may contribute to the persistence of SL9-mediated immune responses in chronically infected individuals. PMID- 16702213 TI - Transduction of a mesenchyme-specific gene periostin into 293T cells induces cell invasive activity through epithelial-mesenchymal transformation. AB - Tumor metastasis is a multistep pathological process involved in the final phase of tumor development. During this process, epithelium-derived tumor cells undergo fibroblast-like transformation, referred to as epithelial-mesenchymal transition (EMT), which contributes to aggressive behavior of tumors. We identify periostin, a mesenchyme-specific gene product, as a contributor to EMT and metastatic potential. Stable expression of a periostin transgene in tumorigenic but non metastatic 293T cells caused cells to undergo fibroblast-like transformation accompanied by increased expressions of vimentin, epidermal growth factor receptor (EGFR), and matrix metalloproteinase-9. The cells expressing ectopic periostin increased cell migration, invasion, and adhesion by 2-9-fold. Invasive characteristics required signaling through integrin alpha(v)beta5 and EGFR. In addition, periostin-engineered 293T cells formed metastases in immunodeficient mice following either cardiac inoculation or injection into mammary fat pad. These data demonstrate an active role for periostin in EMT and metastasis that requires cross-talk between integrin and EGFR signaling pathways. PMID- 16702211 TI - NMR structure of AbhN and comparison with AbrBN: FIRST insights into the DNA binding promiscuity and specificity of AbrB-like transition state regulator proteins. AB - Understanding the molecular mechanisms of transition state regulator proteins is critical, since they play a pivotal role in the ability of bacteria to cope with changing environments. Although much effort has focused on their genetic characterization, little is known about their structural and functional conservation. Here we present the high resolution NMR solution structure of the N terminal domain of the Bacillus subtilis transition state regulator Abh (AbhN), only the second such structure to date. We then compare AbhN to the N-terminal DNA-binding domain of B. subtilis AbrB (AbrBN). This is the first such comparison between two AbrB-like transition state regulators. AbhN and AbrBN are very similar, suggesting a common structural basis for their DNA binding. However, we also note subtle variances between the AbhN and AbrBN structures, which may play important roles in DNA target specificity. The results of accompanying in vitro DNA-binding studies serve to highlight binding differences between the two proteins. PMID- 16702215 TI - In vitro characterization of the interaction between HIV-1 Gag and human lysyl tRNA synthetase. AB - Human immunodeficiency virus type 1 (HIV-1) viral assembly is mediated by multiple protein-protein and protein-nucleic acid interactions. Human tRNA(Lys3) is used as the primer for HIV reverse transcription, and HIV Gag and GagPol are required for packaging of the tRNA into virions. Human lysyl-tRNA synthetase (LysRS) is also specifically packaged into HIV, suggesting a role for LysRS in tRNA packaging. Gag alone is sufficient for packaging of LysRS, and these two proteins have been shown to interact in vitro using glutathione S-transferase pull-down assays. In vitro pull-down assays using truncated constructs have also revealed that residues important for homodimerization of Gag and LysRS are critical for the Gag/LysRS interaction. In this work, we report further in vitro characterization of the interaction between HIV Gag and human LysRS using affinity pull-down assays, fluorescence anisotropy measurements and gel chromatography. An equilibrium binding constant of 310 +/- 80 nM was measured for the Gag/LysRS interaction. We also show that capsid alone binds to LysRS with a similar affinity as full-length Gag. Point mutations that disrupt the homodimerization of LysRS and Gag in vitro do not affect their interaction. These results suggest that dimerization of each protein per se is not required for the interaction but that residues involved in forming the homodimer interfaces contribute to heterodimer formation. Gel chromatography studies further support the formation of a Gag/LysRS heterodimer. PMID- 16702214 TI - Importance of cAMP-response element-binding protein in regulation of expression of the murine cyclic nucleotide phosphodiesterase 3B (Pde3b) gene in differentiating 3T3-L1 preadipocytes. AB - Incubation of 3T3-L1 preadipocytes with isobutylmethylxanthine (IBMX), dexamethasone, and insulin, alone or in combination, demonstrated that IBMX, which increased cAMP-response element-binding protein (CREB) phosphorylation, was the predominant regulator of Pde3b expression. Real time PCR and immunoblotting indicated that in 3T3-L1 preadipocytes, IBMX-stimulated induction of Pde3b mRNA and protein was markedly inhibited by dominant-negative CREB proteins. By transfecting preadipocytes, differentiating preadipocytes, and HEK293A cells with luciferase reporter vectors containing different fragments of the 5'-flanking region of the Pde3b gene, we identified a distal promoter that contained canonical cis-acting cAMP-response elements (CRE) and a proximal, GC-rich promoter region, which contained atypical CRE. Mutation of the CRE sequences dramatically reduced distal promoter activity; H89 inhibited IBMX-stimulated CREB phosphorylation and proximal and distal promoter activities. Distal promoter activity was stimulated by IBMX and phorbol ester (PMA) in Raw264.7 monocytes, but only by IBMX in 3T3-L1 preadipocytes. Chromatin immunoprecipitation analyses with specific antibodies against CREB, phospho-CREB, and CBP/p300 (CREB-binding protein) showed that these proteins associated with both distal and proximal promoters and that interaction of phospho-CREB, the active form of CREB, with both Pde3b promoter regions was increased in IBMX-treated preadipocytes. These results indicate that CRE in distal and proximal promoter regions and activation of CREB proteins play a crucial role in transcriptional regulation of Pde3b expression during preadipocyte differentiation. PMID- 16702216 TI - Exchange of a single amino acid switches the substrate properties of RhoA and RhoD toward glucosylating and transglutaminating toxins. AB - Rho GTPases are the preferred targets of various bacterial cytotoxins, including Clostridium difficile toxins A and B, Clostridium sordellii lethal toxin, the cytotoxic necrotizing factors (CNF1) from Escherichia coli, and the dermonecrotizing toxin (DNT) from Bordetella species. The toxins inactivate or activate specific sets of Rho GTPases by mono-O-glucosylation and deamidation/transglutamination, respectively. Here we studied the structural basis of the recognition of RhoA, which is modified by toxin B, CNF1, and DNT, in comparison with RhoD, which is solely a substrate for lethal toxin. We found that a single amino acid residue in RhoA and RhoD defines the substrate specificity for toxin B and lethal toxin. Change of serine 73 to phenylalanine in RhoA turned RhoA into a substrate for lethal toxin. Accordingly, change of the equivalently positioned phenylalanine 85 in RhoD with serine allowed glucosylation by toxin B. Comparable results were achieved with the Rho-activating and transglutaminating enzymes CNF1 and DNT. Here, amino acid glutamate 64 of RhoA and the equivalent aspartate 76 of RhoD define substrate specificity for CNF1 and DNT, respectively. These data indicate that single amino acid residues located in the switch II region of Rho proteins determine enzyme specificity for diverse bacterial toxins. PMID- 16702217 TI - Resurgent current and voltage sensor trapping enhanced activation by a beta scorpion toxin solely in Nav1.6 channel. Significance in mice Purkinje neurons. AB - Resurgent currents are functionally crucial in sustaining the high frequency firing of cerebellar Purkinje neurons expressing Na(v)1.6 channels. Beta-scorpion toxins, such as CssIV, induce a left shift in the voltage-dependent activation of Na(v)1.2 channels by "trapping" the IIS4 voltage sensor segment. We found that the dangerous Cn2 beta-scorpion peptide induces both the left shift voltage dependent activation and a transient resurgent current only in human Na(v)1.6 channels (among 1.1-1.7), whereas CssIV did not induce the resurgent current. Cn2 also produced both actions in mouse Purkinje cells. These findings suggest that only distinct beta-toxins produce resurgent currents. We suggest that the novel and unique selectivity of Cn2 could make it a model drug to replace deep brain stimulation of the subthalamic nucleus in patients with Parkinson disease. PMID- 16702218 TI - Mechanistic study on the oxidation of anthocyanidin synthase by quantum mechanical calculation. AB - Anthocyanidin synthase (ANS), a member of the 2-oxoglutarate-dependent dioxygenase family in flavonoid biosynthesis, catalyzes the conversion of leucoanthocyanidins (e.g. 2R,3S,4S-cis-leucocyanidin, LCD) to flav-2-en-3,4 diols, a direct precursor of colored anthocyanidins via flavan-3,3,4-triols. The detailed oxygenation mechanism of 2R,3S,4S-cis-LCD to flav-2-en-3,4-diols was investigated using the density functional theory method. An initial model for the calculation was constructed from a structure obtained by a 100-ps molecular dynamics simulation of Arabidopsis ANS under physiological conditions. This model consisted of an LCD molecule as the substrate together with an iron atom, two histidine residues, an aspartic acid residue, a succinate, and an oxygen atom as ligands of the iron atom. The results of the calculation indicated that both the C-3 and C-4 positions of LCD can be oxidized, although C-4 oxidation is preferable. The C-3 oxidation required several steps to form flavan-3,3,4-triol: 1) formation of Fe(III)-OH and a substrate C-3 radical via hydrogen atom abstraction by Fe(IV)=O, 2) formation of a C-3 ketone and a water molecule, 3) addition of OH(-) into the C-3 position of the ketone, and 4) addition of H(+) to form flavan-3,3,4-triol. On the other hand, C-4 oxidation of 2R,3S,4S-cis-LCD resulted in the direct formation of 2R,3R-trans-dihydroquercetin. These results suggest that the oxidation at C-3 of LCD, a key reaction for coloring in anthocyanin biosynthesis, can be regarded as a "side reaction" from the viewpoint of quantum mechanics of enzymatic reactions. Molecular evolutional implications of ANS and related proteins are discussed in terms of reaction dynamics. PMID- 16702219 TI - Liposomes comprising anionic but not neutral phospholipids cause dissociation of Rac(1 or 2) x RhoGDI complexes and support amphiphile-independent NADPH oxidase activation by such complexes. AB - Activation of the phagocyte NADPH oxidase involves the assembly of a membrane localized cytochrome b559 with the cytosolic components p47(phox), p67(phox), p40(phox), and the GTPase Rac (1 or 2). In resting phagocytes, Rac is found in the cytosol as a prenylated protein in the GDP-bound form, associated with the Rho GDP dissociation inhibitor (RhoGDI). In the process of NADPH oxidase activation, Rac is dissociated from RhoGDI and translocates to the membrane, in concert with the other cytosolic components. The mechanism responsible for dissociation of Rac from RhoGDI is poorly understood. We generated Rac(1 or 2) x RhoGDI complexes in vitro from recombinant Rac(1 or 2), prenylated enzymatically, and recombinant RhoGDI, and purified these by anion exchange chromatography. Exposing Rac(1 or 2)(GDP) x RhoGDI complexes to liposomes containing four different anionic phospholipids caused the dissociation of Rac(1 or 2)(GDP) from RhoGDI and its binding to the anionic liposomes. Rac2(GDP) x RhoGDI complexes were more resistant to dissociation, reflecting the lesser positive charge of Rac2. Liposomes consisting of neutral phospholipid did not cause dissociation of Rac(1 or 2) x RhoGDI complexes. Rac1 exchanged to the hydrolysis-resistant GTP analogue, GMPPNP, associated with RhoGDI with lower affinity than Rac1(GDP) and Rac1(GMPPNP) x RhoGDI complexes were more readily dissociated by anionic liposomes. Rac1(GMPPNP) x RhoGDI complexes elicited NADPH oxidase activation in native phagocyte membrane liposomes in the presence of p67(phox), without the need for an anionic amphiphile, as activator. Both Rac1(GDP) x RhoGDI and Rac1(GMPPNP) x RhoGDI complexes elicited amphiphile-independent, p67(phox) dependent NADPH oxidase activation in phagocyte membrane liposomes enriched in anionic phospholipids but not in membrane liposomes enriched in neutral phospholipids. PMID- 16702220 TI - Chondroitin sulfate/dermatan sulfate hybrid chains in the development of cerebellum. Spatiotemporal regulation of the expression of critical disulfated disaccharides by specific sulfotransferases. AB - Chondroitin sulfate/dermatan sulfate (CS/DS) chains regulate the development of the central nervous system in vertebrates. Previously, we demonstrated that CS/DS hybrid chains from embryonic pig brain exhibit neuritogenic and growth factor binding activities, which depended on their IdoUA content defining the DS-like structure. To elucidate the distribution of such functional sugar chains during the development of the brain, in situ hybridization was performed to examine expression of three CS/DS GalNAc 4-O-sulfotransferases, D4ST-1, C4ST-1, and C4ST 2, and a single uronyl 2-O-sulfotransferase (UST) involved in the biosynthesis of DS in addition to CS intermediates. C4ST-1 and C4ST-2 were ubiquitously expressed in the postnatal mouse brain, whereas the expression of D4ST-1 and UST was restricted in the developing cerebellum and culminated at postnatal day 14 as shown by reverse transcriptase-PCR analysis. In situ analysis of the disaccharides of CS/DS in brain sections revealed that the concentration of CS/DS increases 2-fold during development (postnatal day 7 to 7 weeks). The proportions of DS-specific, principal disaccharides, IdoUA-Gal-NAc(4-O-sulfate) (iA) and IdoUA(2-O-sulfate)-GalNAc(4-O-sulfate) (iB), produced by the sequential actions of D4ST-1 and UST, were higher in the CS/DS chains from cerebellum than those from whole brain sections. A dramatic increase (10-fold) in the proportion of iB during development was noteworthy. In contrast, GlcUA/IdoUA(2-O-sulfate)-GalNAc(6 O-sulfate) (D/iD) and GlcUA/IdoUA-GalNAc(4, 6-O-disulfate) (E/iE) decreased to 50 and 30%, respectively, in the developing cerebellum. These results suggest that the IdoUA-containing iA and iB units along with D/iD and E/iE units in the CS/DS hybrid play important roles in the formation of the cerebellar neural network during postnatal brain development. PMID- 16702221 TI - SHAP potentiates the CD44-mediated leukocyte adhesion to the hyaluronan substratum. AB - CD44-hyaluronan (HA) interaction is involved in diverse physiological and pathological processes. Regulation of interacting avidity is well studied on CD44 but rarely on HA. We discovered a unique covalent modification of HA with a protein, SHAP, that corresponds to the heavy chains of inter-alpha-trypsin inhibitor family molecules circulating in blood. Formation of the SHAP.HA complex is often associated with inflammation, a well known process involving the CD44-HA interaction. We therefore examined the effect of SHAP on the CD44-HA interaction mediated lymphocyte adhesion. Under both static and flowing conditions, Hut78 cells (CD44-positive) and CD44-transfected Jurkat cells (originally CD44 negative) adhered preferentially to the immobilized SHAP.HA complex than to HA. The enhanced adhesion is exclusively mediated by the CD44-HA interaction, because it was inhibited by HA, but not IalphaI, and was completely abolished by pretreating the cells with anti-CD44 antibodies. SHAP appears to potentiate the interaction by increasing the avidity of HA to CD44 and altering their distribution on cell surfaces. Large amounts of the SHAP.HA complex accumulate in the hyperplastic synovium of rheumatoid arthritis patients. Leukocytes infiltrated to the synovium were strongly positive for HA, SHAP, and CD44 on their surfaces, suggesting a role for the adhesion-enhancing effect of SHAP in pathogenesis. PMID- 16702222 TI - Conservation of structure and mechanism in primary and secondary transporters exemplified by SiaP, a sialic acid binding virulence factor from Haemophilus influenzae. AB - Extracytoplasmic solute receptors (ESRs) are important components of solute uptake systems in bacteria, having been studied extensively as parts of ATP binding cassette transporters. Herein we report the first crystal structure of an ESR protein from a functionally characterized electrochemical ion gradient dependent secondary transporter. This protein, SiaP, forms part of a tripartite ATP-independent periplasmic transporter specific for sialic acid in Haemophilus influenzae. Surprisingly, the structure reveals an overall topology similar to ATP binding cassette ESR proteins, which is not apparent from the sequence, demonstrating that primary and secondary transporters can share a common structural component. The structure of SiaP in the presence of the sialic acid analogue 2,3-didehydro-2-deoxy-N-acetylneuraminic acid reveals the ligand bound in a deep cavity with its carboxylate group forming a salt bridge with a highly conserved Arg residue. Sialic acid binding, which obeys simple bimolecular association kinetics as determined by stopped-flow fluorescence spectroscopy, is accompanied by domain closure about a hinge region and the kinking of an alpha helix hinge component. The structure provides insight into the evolution, mechanism, and substrate specificity of ESR-dependent secondary transporters that are widespread in prokaryotes. PMID- 16702223 TI - Role of the chaperonin CCT/TRiC complex in G protein betagamma-dimer assembly. AB - Gbetagamma dimer formation occurs early in the assembly of heterotrimeric G proteins. On nondenaturing (native) gels, in vitro translated, (35)S-labeled Ggamma subunits traveled primarily according to their pI and apparently were not associated with other proteins. In contrast, in vitro translated, (35)S-labeled Gbeta subunits traveled at a high apparent molecular mass (approximately 700 kDa) and co-migrated with the chaperonin CCT complex (also called TRiC). Different FLAG-Gbeta isoforms coprecipitated CCT/TRiC to a variable extent, and this correlated with the ability of the different Gbeta subunits to efficiently form dimers with Ggamma. When translated Ggamma was added to translated Gbeta, a new band of low apparent molecular mass (approximately 50 kDa) was observed, which was labeled by either (35)S-labeled Gbeta or Ggamma, indicating that it is a dimer. Formation of the Gbetagamma dimer was ATP-dependent and inhibited by either adenosine 5'-O-(thiotriphosphate) or aluminum fluoride in the presence of Mg(2+). This inhibition led to increased association of Gbeta with CCT/TRiC. Although Ggamma did not bind CCT/TRiC, addition of Ggamma to previously synthesized Gbeta caused its release from the CCT/TRiC complex. We conclude that the chaperonin CCT/TRiC complex binds to and folds Gbeta subunits and that CCT/TRiC mediates Gbetagamma dimer formation by an ATP-dependent reaction. PMID- 16702224 TI - Cytohesin binder and regulator augments T cell receptor-induced nuclear factor of activated T Cells.AP-1 activation through regulation of the JNK pathway. AB - Cytohesin binder and regulator (Cybr; also known as CYTIP, CASP, and PSCDBP) is a cytokine-induced gene preferentially expressed in hematopoietic tissues and in T helper 1 cells. Cybr protein associates with members of the cytohesin family, which are known ADP-ribosylation factors-GDP/GTP exchange factors, and its functions appear to regulate lymphocyte adhesion and cell-cell contact. Here we show that Cybr mRNA and protein levels are increased upon T cell receptor engagement. Cybr expression then influences T cell receptor-dependent signaling events, such as nuclear factor of activated T cells and AP-1 transcriptional activity. In addition, expression of Cybr results in increased T cell receptor mediated activation of the Rho/Rac exchange factor Vav and of the JNK-p38 MAPK signaling pathway. The effects of Cybr on nuclear factor of activated T cells and AP-1 are dependent on MAPK activation, and enhanced activation of this cascade results in cooperation between the two transcription factors in the regulation of gene expression. These findings provide the first evidence that the adaptor protein Cybr not only regulates lymphocyte adhesion and cell-cell interaction but also contributes to the regulation of the signaling cascade and of the genetic program downstream of the T cell receptor. PMID- 16702225 TI - Sequence specificity of SHP-1 and SHP-2 Src homology 2 domains. Critical roles of residues beyond the pY+3 position. AB - A combinatorial phosphotyrosyl (pY) peptide library was screened to determine the amino acid preferences at the pY+4 to pY+6 positions for the four SH2 domains of protein-tyrosine phosphatases SHP-1 and SHP-2. Individual binding sequences selected from the library were resynthesized and their binding affinities and specificities to various SH2 domains were further evaluated by SPR studies, stimulation of SHP-1 and SHP-2 phosphatase activity, and in vitro pulldown assays. These studies reveal that binding of a pY peptide to the N-SH2 domain of SHP-2 is greatly enhanced by a large hydrophobic residue (Trp, Tyr, Met, or Phe) at the pY+4 and/or pY+5 positions, whereas binding to SHP-1 N-SH2 domain is enhanced by either hydrophobic or positively charged residues (Arg, Lys, or His) at these positions. Similar residues at the pY+4 to pY+6 positions are also preferred by SHP-1 and SHP-2 C-SH2 domains, although their influence on the overall binding affinities is much smaller compared with the N-SH2 domains. A structural model was generated to qualitatively interpret the contribution of the pY+4 and pY+5 residues to the overall binding affinity. Examination of pY motifs from known SHP-1 and SHP-2-binding proteins shows that many of the pY motifs contain a hydrophobic or positively charged residue(s) at the pY+4 and pY+5 positions. PMID- 16702226 TI - Engineering of a single conserved amino acid residue of herpes simplex virus type 1 thymidine kinase allows a predominant shift from pyrimidine to purine nucleoside phosphorylation. AB - Studies of herpes simplex virus type 1 (HSV-1) thymidine (dThd) kinase (TK) crystal structures show that purine and pyrimidine bases occupy distinct positions in the active site but approximately the same geometric plane. The presence of a bulky side chain, such as tyrosine at position 167, would not be sterically favorable for pyrimidine or pyrimidine nucleoside analogue binding, whereas purine nucleoside analogues would be less affected because they are located further away from the phenylalanine side chain. Site-directed mutagenesis of the conserved Ala-167 and Ala-168 residues in HSV-1 TK resulted in a wide variety of differential affinities and catalytic activities in the presence of the natural substrate dThd and the purine nucleoside analogue drug ganciclovir (GCV), depending on the nature of the amino acid mutation. A168H- and A167F mutated HSV-1 TK enzymes turned out to have a virtually complete knock-out of dThd kinase activity (at least approximately 4-5 orders of magnitude lower) presumably due to a steric clash between the mutated amino acid and the dThd ring. In contrast, a full preservation of the GCV (and other purine nucleoside analogues) kinase activity was achieved for A168H TK. The enzyme mutants also markedly lost their binding capacity for dThd and showed a substantially diminished feedback inhibition by thymidine 5'-triphosphate. The side chain size at position 168 seems to play a less important role regarding GCV or dThd selectivity than at position 167. Instead, the nitrogen-containing side chains from A168H and A168K seem necessary for efficient ligand discrimination. This explains why A168H-mutated HSV-1 TK fully preserves its GCV kinase activity (Vmax/Km 4-fold higher than wild-type HSV-1 TK), although still showing a severely compromised dThd kinase activity (Vmax/Km 3-4 orders of magnitude lower than wild-type HSV-1 TK). PMID- 16702227 TI - Autophagy contributes to caspase-independent macrophage cell death. AB - Macrophage cell death plays a role in many physiological and pathophysiological conditions. Previous work has shown that macrophages can undergo caspase independent cell death, and this process is associated with Nur77 induction, which is involved in inducing chromatin condensation and DNA fragmentation. Here we show that autophagy is a cytosolic event that controls caspase-independent macrophage cell death. Autophagy was induced in macrophages treated with lipopolysaccharides (LPSs) and the pan-caspase inhibitor benzyloxycarbonyl-Val Ala-Asp (Z-VAD), and the inhibition of autophagy by either chemical inhibitors or by the RNA interference knockdown of beclin (a protein required for autophagic body formation) inhibited caspase-independent macrophage cell death. We also found an increase in poly(ADP-ribose) (PAR) polymerase (PARP) activation and reactive oxygen species (ROS) production in LPS + Z-VAD-treated macrophages, and both are involved in caspase-independent macrophage cell death. We further determined that the formation of autophagic bodies in macrophages occurs downstream of PARP activation, and PARP activation occurs downstream of ROS production. Using macrophages in which receptor-interacting protein 1 (RIP1) was knocked down by small interfering RNA, and macrophages isolated from Toll/interleukin-1 receptor-domain-containing adaptor inducing IFN-beta (TRIF) deficient mice, we found that TRIF and RIP1 function upstream of ROS production in LPS + Z-VAD-treated macrophages. We also found that Z-VAD inhibits LPS-induced RIP1 cleavage, which may contribute to ROS over-production in macrophages. This paper reveals that TRIF, RIP1, and ROS production, as well as PARP activation, are involved in inducing autophagy, which contributes to caspase-independent macrophage cell death. PMID- 16702228 TI - Polychlorinated biphenyl-induced reduction of dopamine transporter expression as a precursor to Parkinson's disease-associated dopamine toxicity. AB - Epidemiological and laboratory studies have suggested that exposure to polychlorinated biphenyls (PCBs) may be a risk factor for Parkinson's disease. The purpose of this study was to examine the potential mechanisms by which PCBs may disrupt normal functioning of the nigrostriatal dopamine (DA) system. We utilized an environmentally relevant exposure of PCBs (7.5 or 15 mg/kg/day Aroclor 1,254:1,260 for 30 days by oral gavage) to identify early signs of damage to the DA system. This dosing regimen, which resulted in PCB levels similar to those found in human brain samples, did not cause overt degeneration to the DA system as shown by a lack of change in striatal DA levels or tyrosine hydroxylase levels. However, we did observe a dramatic dose-dependent decrease in striatal dopamine transporter (DAT) levels. The observed reductions appear to be specific to the DAT populations located in the striatum, as no change was observed in other dopaminergic brain regions or to other neurotransmitter transporters present in the striatum. These data demonstrate that PCB tissue concentrations similar to those found in postmortem human brain specifically disrupt DA transport, which acts as a precursor to subsequent damage to the DA system. Furthermore, DAT imaging may be useful in evaluating alterations in brain function in human populations exposed to PCBs. PMID- 16702229 TI - Outlier sums for differential gene expression analysis. AB - We propose a method for detecting genes that, in a disease group, exhibit unusually high gene expression in some but not all samples. This can be particularly useful in cancer studies, where mutations that can amplify or turn off gene expression often occur in only a minority of samples. In real and simulated examples, the new method often exhibits lower false discovery rates than simple t-statistic thresholding. We also compare our approach to the recent cancer profile outlier analysis proposal of Tomlins and others (2005). PMID- 16702230 TI - Semaphorin 4D/Plexin-B1-mediated R-Ras GAP activity inhibits cell migration by regulating beta(1) integrin activity. AB - Plexins are cell surface receptors for semaphorins and regulate cell migration in many cell types. We recently reported that the semaphorin 4D (Sema4D) receptor Plexin-B1 functions as a GTPase-activating protein (GAP) for R-Ras, a member of Ras family GTPases implicated in regulation of integrin activity and cell migration. We characterized the role of R-Ras downstream of Sema4D/Plexin-B1 in cell migration. Activation of Plexin-B1 by Sema4D suppressed the ECM-dependent R Ras activation, R-Ras-mediated phosphatydylinositol 3-kinase activation, and beta(1) integrin activation through its R-Ras GAP domain, leading to inhibition of cell migration. In addition, inactivation of R-Ras by overexpression of the R Ras-specific GAP or knockdown of R-Ras by RNA interference was sufficient for suppressing beta(1) integrin activation and cell migration in response to the ECM stimulation. Thus, we conclude that R-Ras activity is critical for ECM-mediated beta(1) integrin activation and cell migration and that inactivation of R-Ras by Sema4D/Plexin-B1-mediated R-Ras GAP activity controls cell migration by modulating the activity of beta(1) integrins. PMID- 16702231 TI - Optimization of WAVE2 complex-induced actin polymerization by membrane-bound IRSp53, PIP(3), and Rac. AB - WAVE2 activates the actin-related protein (Arp) 2/3 complex for Rac-induced actin polymerization during lamellipodium formation and exists as a large WAVE2 protein complex with Sra1/PIR121, Nap1, Abi1, and HSPC300. IRSp53 binds to both Rac and Cdc42 and is proposed to link Rac to WAVE2. We found that the knockdown of IRSp53 by RNA interference decreased lamellipodium formation without a decrease in the amount of WAVE2 complex. Localization of WAVE2 at the cell periphery was retained in IRSp53 knockdown cells. Moreover, activated Cdc42 but not Rac weakened the association between WAVE2 and IRSp53. When we measured Arp2/3 activation in vitro, the WAVE2 complex isolated from the membrane fraction of cells was fully active in an IRSp53-dependent manner but WAVE2 isolated from the cytosol was not. Purified WAVE2 and purified WAVE2 complex were activated by IRSp53 in a Rac dependent manner with PIP(3)-containing liposomes. Therefore, IRSp53 optimizes the activity of the WAVE2 complex in the presence of activated Rac and PIP(3). PMID- 16702232 TI - The clathrin adaptor complex 1 directly binds to a sorting signal in Ste13p to reduce the rate of its trafficking to the late endosome of yeast. AB - Yeast trans-Golgi network (TGN) membrane proteins maintain steady-state localization by constantly cycling to and from endosomes. In this study, we examined the trafficking itinerary and molecular requirements for delivery of a model TGN protein A(F-->A)-alkaline phosphatase (ALP) to the prevacuolar/endosomal compartment (PVC). A(F-->A)-ALP was found to reach the PVC via early endosomes (EEs) with a half-time of approximately 60 min. Delivery of A(F-->A)-ALP to the PVC was not dependent on either the GGA or adaptor protein 1 (AP-1) type of clathrin adaptors, which are thought to function in TGN to PVC and TGN to EE transport, respectively. Surprisingly, in cells lacking the function of both GGA and AP-1 adaptors, A(F-->A)-ALP transport to the PVC was dramatically accelerated. A 12-residue cytosolic domain motif of A(F-->A)-ALP was found to mediate direct binding to AP-1 and was sufficient to slow TGN-->EE-->PVC trafficking. These results suggest a model in which this novel sorting signal targets A(F-->A)-ALP into clathrin/AP-1 vesicles at the EE for retrieval back to the TGN. PMID- 16702233 TI - NDC1: a crucial membrane-integral nucleoporin of metazoan nuclear pore complexes. AB - POM121 and gp210 were, until this point, the only known membrane-integral nucleoporins (Nups) of vertebrates and, thus, the only candidate anchors for nuclear pore complexes (NPCs) within the nuclear membrane. In an accompanying study (Stavru et al.), we provided evidence that NPCs can exist independently of POM121 and gp210, and we predicted that vertebrate NPCs contain additional membrane-integral constituents. We identify such an additional membrane protein in the NPCs of mammals, frogs, insects, and nematodes as the orthologue to yeast Ndc1p/Cut11p. Human NDC1 (hNDC1) likely possesses six transmembrane segments, and it is located at the nuclear pore wall. Depletion of hNDC1 from human HeLa cells interferes with the assembly of phenylalanine-glycine repeat Nups into NPCs. The loss of NDC1 function in Caenorhabditis elegans also causes severe NPC defects and very high larval and embryonic mortality. However, it is not ultimately lethal. Instead, homozygous NDC1-deficient worms can be propagated. This indicates that none of the membrane-integral Nups is universally essential for NPC assembly, and suggests that NPC biogenesis is an extremely fault-tolerant process. PMID- 16702234 TI - Nuclear pore complex assembly and maintenance in POM121- and gp210-deficient cells. AB - So far, POM121 and gp210 are the only known anchoring sites of vertebrate nuclear pore complexes (NPCs) within the lipid bilayer of the nuclear envelope (NE) and, thus, are excellent candidates for initiating the NPC assembly process. Indeed, we demonstrate that POM121 can recruit several nucleoporins, such as Nup62 or Nup358, to ectopic assembly sites. It thus appears to act as a nucleation site for the assembly of NPC substructures. Nonetheless, we observed functional NPCs and intact NEs in severely POM121-depleted cells. Double knockdowns of gp210 and POM121 in HeLa cells, as well as depletion of POM121 from human fibroblasts, which do not express gp210, further suggest that NPCs can assemble or at least persist in a POM121- and gp210-free form. This points to extensive redundancies in protein-protein interactions within NPCs and suggests that vertebrate NPCs contain additional membrane-integral nucleoporins for anchorage within the lipid bilayer of the NE. In Stavru et al., we describe such an additional transmembrane nucleoporin as the metazoan orthologue of yeast Ndc1p. PMID- 16702236 TI - Riding the DUBway: regulation of protein trafficking by deubiquitylating enzymes. AB - Ubiquitylation is a key regulator of protein trafficking, and much about the functions of ubiquitin ligases, which add ubiquitin to substrates in this regulation, has recently come to light. However, a clear understanding of ubiquitin-dependent protein localization cannot be achieved without knowledge of the role of deubiquitylating enzymes (DUBs). DUBs, by definition, function downstream in ubiquitin pathways and, as such, have the potential to be the final editors of protein ubiquitylation status, thus determining substrate fate. This paper assimilates the current evidence concerning the substrates and activities of DUBs that regulate protein trafficking. PMID- 16702237 TI - Radiation exposure during travelling in Malaysia. AB - Absorbed dose rates in vehicles during travelling by different modes of transport in Malaysia were measured. Radiation levels measured on roads in Peninsular Malaysia were within a broad range, i.e. between 36 and 1560 nGy h(-1). The highest reading, recorded while travelling near monazite and zircon mineral dumps, was 13 times the mean environmental radiation level of Malaysia. It is evident that radioactive material dumps on the roadsides can influence the radiation level on the road. The absorbed dose rates measured while travelling on an ordinary train were between 60 and 350 nGy h(-1). The highest reading was measured when the train passed a tunnel built through a granite rock hill. The measurement during sea travelling by ferries gave the lowest radiation level owing to merely cosmic radiation at the sea level. PMID- 16702235 TI - M line-deficient titin causes cardiac lethality through impaired maturation of the sarcomere. AB - Titin, the largest protein known to date, has been linked to sarcomere assembly and function through its elastic adaptor and signaling domains. Titin's M-line region contains a unique kinase domain that has been proposed to regulate sarcomere assembly via its substrate titin cap (T-cap). In this study, we use a titin M line-deficient mouse to show that the initial assembly of the sarcomere does not depend on titin's M-line region or the phosphorylation of T-cap by the titin kinase. Rather, titin's M-line region is required to form a continuous titin filament and to provide mechanical stability of the embryonic sarcomere. Even without titin integrating into the M band, sarcomeres show proper spacing and alignment of Z discs and M bands but fail to grow laterally and ultimately disassemble. The comparison of disassembly in the developing and mature knockout sarcomere suggests diverse functions for titin's M line in embryonic development and the adult heart that not only involve the differential expression of titin isoforms but also of titin-binding proteins. PMID- 16702238 TI - Unexpectedly high activity of 228Th in excretion samples following consumption of Brazil nuts. AB - A worker provided a routine faecal sample for plutonium and americium analysis. In the course of this analysis 500 mBq of (228)Th was discovered. There seemed no credible occupational route for intake of thorium. Further investigation revealed that the worker consumed approximately 25 g d(-1) of nuts, including Brazil nuts. A sample of these nuts was analysed and found to contain activities of (228)Th in sufficient quantity to account for the faecal activity. However, follow-up urine samples taken from the worker showed 0.6-0.7 mBq of (228)Th. The intake of (228)Th via nuts is insufficient to account for this activity in urine. However, it is likely that the intake of (228)Th was accompanied by similar activity of the parent (228)Ra, and biokinetic calculations show that decay of (228)Ra in vivo would produce sufficient (228)Th to account for the observed urine activity. PMID- 16702239 TI - Gamma activity and radiation dose in concrete building blocks used for construction of dwellings in Jos, Nigeria. AB - Radioactivity concentrations of concrete building block samples made and used in Jos, Nigeria, were determined using a gamma ray spectrometry method. The mean values of the radioactivity concentrations were calculated as 66, 126 and 589 Bq kg(-1) for (226)Ra, (232)Th and (40)K, respectively. The radium equivalent activities ranged between 131 and 712 Bq kg(-1), with six of the samples exceeding the UNSCEAR reported maximum permissible level of 370 Bq kg(-1). The mean radium equivalent activity (292 Bq kg(-1)) is higher than the values available in the literature for some countries by factors ranging between 2 and 5, a situation that can be traced to the practice of sourcing sand aggregates from mining sites. The indoor annual effective dose for a dwelling of dimension 3.6 x 3.6 x 3.0 m(3) was calculated as 0.81 mSv. PMID- 16702240 TI - Analysis of radon concentration in drinking water in Baoji (China) and the associated health effects. AB - This paper presents the results of radon concentration measurements in drinking water from the municipal water supply system and private wells located in Baoji, China. The measurements were carried out on 69 samples. The mean values of tap water and well water were found to be 12 kBq m(-3) with a maximum of 18 kBq m(-3) and 41 kBq m(-3) with a maximum of 127 kBq m(-3), respectively. The well water samples obtained from different depth-well (water-bearing levels), i.e. shallow well (well depth under 10 m) water, middle well (well depth 10-30 m) water and deep well water, have respective mean values of 24, 34 and 56 kBq m(-3). The contributions of the observed radon concentration in drinking water to indoor radon account for 2.8-13.2% of the mean value of Shaanxi indoor radon concentration and the effective dose to the dweller owing to inhalation of radon emanating from household water is 0.03-0.14 mSv y(-1). PMID- 16702241 TI - Batch homogeneity of LiF(Mg,Cu,P)-GR200 and LiF(Mg,Cu,P)-MCP-NS TL detectors for use as extremity dosemeters at ENEA personal dosimetry service. AB - The results of a study of two commercially available LiF(Mg,Cu,P) TL materials, a GR200 detector and a MCP-Ns thin detector, are described in order to use these phosphors for individual monitoring for the extremities. After a dosimetry system has been type tested, the implementation routine is not straightforward. Additional tests and software modification are needed to make the routine system work comply with the type test results. Not often can literature be found on the steps required to implement the results in a routine study. This paper reports the results of the individual calibration of about 15 000 extremity dosemeters, 12 000 containing a GR200 detector and 3000 an MCP-Ns thin detector. It describes the experimental procedure followed in order to assure reproducibility and stability of the results with proper accuracy and reliability. In particular, this is the first time that results on homogeneity of such a large batch of MCP Ns detectors are reported. PMID- 16702242 TI - Calculation and properties of trap structural functions for various spatially correlated systems. AB - Thermoluminescence (TL) kinetics in spatially inhomogeneous systems can be studied by various Monte Carlo algorithms. Recently, a new analytical approach was suggested for the isolated cluster model. The theory is based on the concept of trap structural functions (TSFs). TSFs depend solely on topological properties of solids. Therefore, knowing TSFs for traps and recombination centres it is possible to calculate TL for various parameters, e.g. different heating schemes and different energy configurations. This paper presents some properties and methods of calculation of TSFs. Structural character of TSFs is verified numerically. PMID- 16702243 TI - Precise dose evaluation using a commercial phototransistor as a radiation detector. AB - An experimental arrangement and a circuitry based on an NPN phototransistor-type silicon radiation detector have been used for evaluating the X-ray beam dose in the diagnostic range. The circuitry was built to allow alteration of the electric field in the phototransistor internal structure, with some devices that have an available base connection. By changing the transistor base bias it is possible to alter its operation point to obtain a response gain from the selected photon energy range. In this way we have made an electronic energy-domain discretisation and we are investigating a model to calculate the dose contribution from each energy discretised into 10 keV steps. The method has been tested in filtered radiation beams generated from an HF-160 Pantak X-ray unit and compared with the usual dosimetry method. Our results have demonstrated that it is possible to make such a dose deconvolution from 40 to 140 keV energies by controlling the phototransistor base bias properly. PMID- 16702244 TI - Thermoluminescence response of new KCl(X)Br(1-X):EuCl3 sintered phosphors exposed to beta and gamma radiation. AB - Alkali halides crystals have been the subject of intense research for an understanding of their radiation-induced defects and luminescence properties. They exhibit noteworthy thermoluminescence (TL) properties when exposed to ionising radiation. Currently, these materials are grown employing expensive and rather complicated techniques. In this work, the results on the TL properties of new alkali halides phosphors fabricated by a simple and inexpensive procedure are presented. The samples were made by mixing KCl, KBr and EuCl3 salts, and compressing them at a pressure of 3.2 x 10(7) Pa during 3 min, followed by sintering at 700 degrees C during 24 h under air atmosphere. The dosimetric response of the samples showed an increase with radiation dose in the 1.5-20.0 Gy dose range for beta and gamma radiation. The TL glow curves in sintered samples presented significant differences in their peak structures compared with monocrystalline samples, indicating that the nature of the trapping states and the recombination mechanisms may be different. PMID- 16702245 TI - Optical and dosimetric properties of zircon. AB - Irradiation effects were investigated in zircon crystals by methods of optical absorption and luminescence. Special attention was given to the effects of vacuum ultraviolet (VUV) radiation. The same main thermoluminescence (TL) peaks with the same thermal activation energies appeared after VUV as after X- or beta irradiation, indicating that the same traps were induced by the different irradiations. TL excitation spectra in the VUV showed an increase <220 nm and maxima near 190 and 140 nm. Excitation spectra of phototransferred TL (PTTL) and optically stimulated luminescence (OSL) were also measured. Most TL emission bands also appeared in the X-luminescence, PTTL and OSL. Dosimetric properties such as the TL radiation sensitivity, thermal stability of radiation-induced defects and TL dose dependence were also investigated. The radiation sensitivity of zircon was by an order of magnitude lower than that of TLD-100. The 355 K TL peak showed linear dose dependence only up to approximately 500 Gy and the 520 K peak up to approximately 1800 Gy. PMID- 16702246 TI - Dosimetry at the Portuguese research reactor using thermoluminescence measurements and Monte Carlo calculations. AB - This work presents an extensive study on Monte Carlo radiation transport simulation and thermoluminescent (TL) dosimetry for characterising mixed radiation fields (neutrons and photons) occurring in nuclear reactors. The feasibility of these methods is investigated for radiation fields at various locations of the Portuguese Research Reactor (RPI). The performance of the approaches developed in this work is compared with dosimetric techniques already existing at RPI. The Monte Carlo MCNP-4C code was used for a detailed modelling of the reactor core, the fast neutron beam and the thermal column of RPI. Simulations using these models allow to reproduce the energy and spatial distributions of the neutron field very well (agreement better than 80%). In the case of the photon field, the agreement improves with decreasing intensity of the component related to fission and activation products. (7)LiF:Mg,Ti, (7)LiF:Mg,Cu,P and Al(2)O(3):Mg,Y TL detectors (TLDs) with low neutron sensitivity are able to determine photon dose and dose profiles with high spatial resolution. On the other hand, (nat)LiF:Mg,Ti TLDs with increased neutron sensitivity show a remarkable loss of sensitivity and a high supralinearity in high-intensity fields hampering their application at nuclear reactors. PMID- 16702247 TI - The Third International Intercomparison on EPR Tooth Dosimetry: part 2, final analysis. AB - The objective of the Third International Intercomparison on EPR Tooth Dosimetry was to evaluate laboratories performing tooth enamel dosimetry <300 mGy. Final analysis of results included a correlation analysis between features of laboratory dose reconstruction protocols and dosimetry performance. Applicability of electron paramagnetic resonance (EPR) tooth dosimetry at low dose was shown at two applied dose levels of 79 and 176 mGy. Most (9 of 12) laboratories reported the dose to be within 50 mGy of the delivered dose of 79 mGy, and 10 of 12 laboratories reported the dose to be within 100 mGy of the delivered dose of 176 mGy. At the high-dose tested (704 mGy) agreement within 25% of the delivered dose was found in 10 laboratories. Features of EPR dose reconstruction protocols that affect dosimetry performance were found to be magnetic field modulation amplitude in EPR spectrum recording, EPR signal model in spectrum deconvolution and duration of latency period for tooth enamel samples after preparation. PMID- 16702248 TI - The intercomparison of cosmic rays with heavy ion beams at NIRS (ICCHIBAN) project. AB - The ICCHIBAN-2 experiment, the first dedicated to the ground-based intercomparison of passive space dosemeters, was carried out between 23 May and 28 May 2002 at the National Institute of Radiological Sciences in Chiba, Japan. The primary objective of the ICCHIBAN-2 experiment was to intercompare the response of passive dosemeters used in space crew dosimetry to monoenergetic heavy ions of charge and energy spanning a significant portion of the galactic cosmic ray (GCR) spectrum. During the ICCHIBAN-2 experiment, dosemeters from 12 different laboratories in 9 countries were irradiated under identical conditions to heavy ion beams of 150 MeV n(-1) (4)He, 400 MeV n(-1) (12)C, 490 MeV n(-1) (28)Si and 500 MeV n(-1) (56)Fe at the NIRS Heavy Ion Medical Accelerator. PMID- 16702249 TI - Ductus arteriosus aneurysm with community-acquired methicillin-resistant Staphylococcus aureus infection and spontaneous rupture: a potentially fatal quandary. AB - We present the case of a 6-month-old previously healthy girl who presented with high fever, labored breathing, and an enlarged cardiac silhouette on her chest radiograph. Comprehensive evaluation discovered a ductus arteriosus aneurysm and pericardial effusion with methicillin-resistant Staphylococcus aureus bacteremia. Despite pericardiocentesis and appropriate intravenous antibiotics, there was rapid enlargement of the aneurysm and accumulation of echogenic material within the ductus arteriosus aneurysm. Infected aneurysm rupture was identified during emergency surgery. This infant also had vocal cord paresis, a likely complication of the surgery. The clinical course, diagnosis, and treatment of this patient are discussed. Infection of a ductus arteriosus or an infected ductal arteriosus aneurysm is a rare and potentially fatal clinical entity. In the era of increasing community-acquired methicillin-resistant S aureus infections, this is a diagnosis that requires a high index of suspicion. PMID- 16702250 TI - Microstream capnography improves patient monitoring during moderate sedation: a randomized, controlled trial. AB - BACKGROUND: Investigative efforts to improve monitoring during sedation for patients of all ages are part of a national agenda for patient safety. According to the Institute of Medicine, recent technological advances in patient monitoring have contributed to substantially decreased mortality for people receiving general anesthesia in operating room settings. Patient safety has not been similarly targeted for the several million children annually in the United States who receive moderate sedation without endotracheal intubation. Critical event analyses have documented that hypoxemia secondary to depressed respiratory activity is a principal risk factor for near misses and death in this population. Current guidelines for monitoring patient safety during moderate sedation in children call for continuous pulse oximetry and visual assessment, which may not detect alveolar hypoventilation until arterial oxygen desaturation has occurred. Microstream capnography may provide an "early warning system" by generating real time waveforms of respiratory activity in nonintubated patients. OBJECTIVE: The aim of this study was to determine whether intervention based on capnography indications of alveolar hypoventilation reduces the incidence of arterial oxygen desaturation in nonintubated children receiving moderate sedation for nonsurgical procedures. PARTICIPANTS AND METHODS: We included 163 children undergoing 174 elective gastrointestinal procedures with moderate sedation in a pediatric endoscopy unit in a randomized, controlled trial. All of the patients received routine care, including 2-L supplemental oxygen via nasal cannula. Investigators, patients, and endoscopy staff were blinded to additional capnography monitoring. In the intervention arm, trained independent observers signaled to clinical staff if capnograms indicated alveolar hypoventilation for >15 seconds. In the control arm, observers signaled if capnograms indicated alveolar hypoventilation for >60 seconds. Endoscopy nurses responded to signals in both arms by encouraging patients to breathe deeply, even if routine patient monitoring did not indicate a change in respiratory status. OUTCOME MEASURES: Our primary outcome measure was patient arterial oxygen desaturation defined as a pulse oximetry reading of <95% for >5 seconds. Secondary outcome measures included documented assessments of abnormal ventilation, termination of the procedure secondary to concerns for patient safety, as well as other more rare adverse events including need for bag mask ventilation, sedation reversal, or seizures. RESULTS: Children randomly assigned to the intervention arm were significantly less likely to experience arterial oxygen desaturation than children in the control arm. Two study patients had documented adverse events, with no procedures terminated for patient safety concerns. Intervention and control patients did not differ in baseline characteristics. Endoscopy staff documented poor ventilation in 3% of all procedures and no apnea. Capnography indicated alveolar hypoventilation during 56% of procedures and apnea during 24%. We found no change in magnitude or statistical significance of the intervention effect when we adjusted the analysis for age, sedative dose, or other covariates. CONCLUSIONS: The results of this controlled effectiveness trial support routine use of microstream capnography to detect alveolar hypoventilation and reduce hypoxemia during procedural sedation in children. In addition, capnography allowed early detection of arterial oxygen desaturation because of alveolar hypoventilation in the presence of supplemental oxygen. The current standard of care for monitoring all patients receiving sedation relies overtly on pulse oximetry, which does not measure ventilation. Most medical societies and regulatory organizations consider moderate sedation to be safe but also acknowledge serious associated risks, including suboptimal ventilation, airway obstruction, apnea, hypoxemia, hypoxia, and cardiopulmonary arrest. The results of this controlled trial suggest that microstream capnography improves the current standard of care for monitoring sedated children by allowing early detection of respiratory compromise, prompting intervention to minimize hypoxemia. Integrating capnography into patient monitoring protocols may ultimately improve the safety of nonintubated patients receiving moderate sedation. PMID- 16702251 TI - Clinical manifestations and risk factors of children receiving triple ventilating tube insertions for treatment of recurrent otitis media with effusion. AB - OBJECTIVE: Our goal was to determine risk factors for children receiving additional ventilating tube insertions after initial tube insertion and to determine the changes of clinical manifestations in children who have undergone 3 ventilating tube insertion procedures. STUDY DESIGN: We retrospectively analyzed medical and operation records of 423 young patients who had ventilating tube insertion because of chronic otitis media with effusion from January 1993 to December 1998. The single-operation group included patients who had 1 ventilating tube insertion only, and the triple-operation group included patients who received ventilating tube insertion 3 times because of recurring chronic otitis media with effusion. RESULTS: At the first operation, there were significant differences between the single- and triple-operation groups in mean age, the proportion who received a concurrent adenoidectomy, the mean indwelling period of the first ventilating tube, the proportion who developed postoperative otorrhea within 1 month, and the proportion who had early extrusion of the ventilating tube within 3 months of surgery. In the triple-operation group, the accumulated number of adenoidectomies, the indwelling period of the ventilating tube, and the time interval before subsequent ventilating tube insertion after ventilating tube extrusion significantly increased as ventilating tube insertion procedures were performed repeatedly. Although there was no difference when compared with the single-operation group, the proportion of glue-like effusion significantly decreased as ventilating tube insertion procedures were performed repeatedly. There were no significant differences between the single- and triple-operation groups in male/female ratio, site of ventilating tube insertion, and the proportion of patients with glue-like effusion at the first ventilating tube insertion. CONCLUSIONS: The probability of receiving additional ventilating tube insertion because of recurrent otitis media with effusion significantly increased in younger patients at the time of first ventilating tube insertion. The concurrent adenoidectomy, duration of the ventilating tube, postoperative otorrhea within 1 month, and early extrusion of the ventilating tube also influenced the probability of additional ventilating tube insertion. PMID- 16702252 TI - Invited Review: The role of cow, pathogen, and treatment regimen in the therapeutic success of bovine Staphylococcus aureus mastitis. AB - Staphylococcus aureus is an important cause of udder infections in dairy herds. Both lactational and dry cow therapy are part of Staph. aureus control programs. Reported cure rates for Staph. aureus mastitis vary considerably. The probability of cure depends on cow, pathogen, and treatment factors. Cure rates decrease with increasing age of the cow, increasing somatic cell count, increasing duration of infection, increasing bacterial colony counts in milk before treatment, and increasing number of quarters infected. Staphylococcus aureus mastitis in hind quarters has a low cure rate compared with front quarters. Antimicrobial treatment of intramammary infections with penicillin-resistant Staph. aureus strains results in a lower cure rate for treatment with either beta-lactam or non beta-lactam antibiotics. Other strain-specific factors may affect the probability of cure but routine diagnostic methods for use in bacteriology laboratories or veterinary practices are not yet available. The most important treatment factor affecting cure is treatment duration. Increased duration of treatment is associated with increased chance of cure. Economically, extended treatment is not always justified, even when indirect effects of treatment such as prevention of contagious transmission are taken into consideration. Usefulness of treatment trials could be improved by standardization of case definitions, consideration of host and strain factors, and sufficient statistical power. Treatment of young animals with penicillin-sensitive Staph. aureus infections is often justified based on bacteriological cure and economic outcome, whereas treatment of older animals, chronic infections, or penicillin-resistant isolates should be discouraged. PMID- 16702253 TI - Invited review: Culling: nomenclature, definitions, and recommendations. AB - Replacing cows on a dairy is a major cost of operation. There is a need for the industry to adopt a more standardized approach to reporting the rate at which cows exit from the dairy, and to reporting the reasons why cows are replaced and their destination as they exit the dairy. Herd turnover rate is recommended as the preferred term for characterizing the cows exiting a dairy, in preference to herd replacement rate, culling rate, or percent exiting, all of which have served as synonyms. Herd turnover rate should be calculated as the number of cows that exit in a defined period divided by the animal time at risk for the population being characterized. The terms voluntary and involuntary culling suffer from problems of definition and their use should be discouraged. Destination should be recorded for all cows that exit the dairy and opportunities to record one or more reasons for exiting should be provided by management systems. Comparing reported reasons between dairies requires considerable caution because of differences in case definitions and recording methods. Relying upon culling records to monitor disease has been and will always be an ineffective management strategy. Dairies are encouraged to record and monitor disease events and reproductive performance and use this information as the basis for management efforts aimed at reducing the need to replace cows. PMID- 16702254 TI - Stability of casein micelles cross-linked by transglutaminase. AB - In this study, caseins micelles were internally cross-linked using the enzyme transglutaminase (TGase). The integrity of the micelles was examined on solubilization of micellar calcium phosphate (MCP) or on disruption of hydrophobic interactions and breakage of hydrogen bonds. The level of monomeric caseins, determined electrophoretically, decreased with increasing time of incubation with TGase at 30 degrees C; after incubation for 24 h, no monomeric beta- or kappa-caseins were detected, whereas only a small level of monomeric alphaS1-casein remained, suggesting near complete intramicellar cross-linking. The ability of casein micelles to maintain structural integrity on disruption of hydrophobic interactions (using urea, sodium dodecyl sulfate, or heating in the presence of ethanol), solubilization of MCP (using the calcium-chelating agent trisodium citrate) or high-pressure treatment was estimated by measurement of the L*-value of milk; i.e., the amount of back-scattered light. The amount of light scattered by casein micelles in noncross-linked milk was reduced by >95% on complete disruption of hydrophobic interactions or complete solubilization of MCP; treatment of milk with TGase increased the stability of casein micelles against disruption by all methods studied and stability increased progressively with incubation time. After 24 h of cross-linking, reductions in the extent of light scattering were still apparent in the presence of high levels of dissociating agents, possibly through citrate-induced removal of MCP nanoclusters from the micelles, or urea- or sodium dodecyl sulfate-induced increases in solvent refractive index, which reduce the extent of light-scattering. PMID- 16702255 TI - Microfiltration of butter serum upon casein micelle destabilization. AB - The gross composition of butter serum, the aqueous phase of butter, is comparable to that of buttermilk, except that it has a higher content of material derived from the milk fat globule membrane (MFGM). As such, butter serum is a good source for further purification of MFGM material. The purified fraction could be of interest for its emulsifying and nutritional properties. The effect of sodium citrate and ethanol on the dissociation of butter serum casein micelles, and their effect on casein retention upon tangential microfiltration were investigated. Optimal conditions of casein micelle dissociation were assessed by using an experimental design (response surface full central composite orthogonal design) with temperature and ethanol or sodium citrate concentration as design variables and the Hunter L* value as response variable. For both dissociating agents, a highly significant reduced quadratic model was fit to the data. Microfiltration tests were performed on pure butter serum, and on butter serum in the presence of sodium citrate, under optimal dissociation conditions (50 degrees C, 80 mM). A cellulose acetate membrane with a pore size of 0.15 microm was used. From the filtration curves and fouling coefficients it was clear that the addition of sodium citrate improved the permeation flux, and minimized fouling. All fractions were analyzed for dry matter, protein, lactose, lipid, and polar lipid contents. The protein fraction was further characterized by sodium dodecyl sulfate-PAGE. It was shown that sodium citrate greatly enhanced casein transmission through the membrane, but at the expense of substantial losses of polar lipids. PMID- 16702256 TI - Utilization of fourier transform infrared spectroscopy for measurement of organic phosphorus and bound calcium in cheddar cheese. AB - The methods available for measuring organic P and bound Ca in cheese are either cumbersome or involve dilution of the cheese. Dilution of the cheese can lead to erroneous results, particularly in the case of bound Ca. Hence, the objective of this study was to evaluate the feasibility of Fourier transform infrared (FTIR) spectroscopy for direct measurement of organic P and bound Ca in Cheddar cheese. Two hundred sixteen samples of cheese were analyzed for protein-bound organic P, bound Ca using a water-extraction based method, and buffering curves. Additionally, the infrared spectra of the cheeses were collected between 4,000 and 650 cm(-1), at a resolution of 4 cm(-1), and 256 scans per sample. The spectral shifts in the infrared region from 1,050 to 900 cm(-1), in addition to the measured concentrations of organic P, bound Ca, and buffering peak area at pH 5.1, were used to develop calibration models using partial least squares (PLS) regression analysis. The spectral region of 956 to 946 cm(-1) correlated with the measured concentrations of organic P and the overall PLS model had a correlation (R2) of 0.76 between the predicted and measured concentrations. The spectral region at approximately 980 cm(-1) was correlated with the measured concentrations of bound Ca, and the overall PLS model had a correlation (R2) of 0.70 between the predicted and measured concentrations. A similar spectral region at approximately 980 cm(-1) was also correlated with the measured buffering peak areas and the overall PLS model had a correlation (R2) of 0.64 between the predicted and measured peak areas. A linear regression analysis between the bound Ca and buffering peak area demonstrated that bound Ca was correlated (R2 = 0.73) with buffering peak area. This study demonstrates that FTIR can be used to measure organic P in cheeses. It also has the potential to be used for measuring bound Ca in undiluted cheeses, and for prediction of the buffering capacity of cheese. PMID- 16702257 TI - Comparing the fatty acid composition of organic and conventional milk. AB - During a 12-mo longitudinal study, bulk-tank milk was collected each month from organic (n = 17) and conventional (n = 19) dairy farms in the United Kingdom. All milk samples were analyzed for fatty acid (FA) content, with the farming system type, herd production level, and nutritional factors affecting the FA composition investigated by use of mixed model analyses. Models were constructed for saturated fatty acids, the ratio of polyunsaturated fatty acids (PUFA) to monounsaturated fatty acids, total n-3 FA, total n-6 FA, conjugated linoleic acid, and vaccenic acid. The ratio of n-6:n-3 FA in both organic and conventional milk was also compared. Organic milk had a higher proportion of PUFA to monounsaturated fatty acids and of n-3 FA than conventional milk, and contained a consistently lower n-6:n-3 FA ratio (which is considered beneficial) compared with conventional milk. There was no difference between organic and conventional milk with respect to the proportion of conjugated linoleic acid or vaccenic acid. A number of factors other than farming system were identified which affected milk FA content including month of year, herd average milk yield, breed type, use of a total mixed ration, and access to fresh grazing. Thus, organic dairy farms in the United Kingdom produce milk with a higher PUFA content, particularly n-3 FA, throughout the year. However, knowledge of the effects of season, access to fresh grazing, or use of specific silage types could be used by producers to enhance the content of beneficial FA in milk. PMID- 16702258 TI - Factors affecting growth factor activity in goat milk. AB - Growth factors that are present in goat milk may be responsible for its beneficial effects on the digestive system as described in ancient Chinese medical texts. To develop a nutraceutical product rich in growth factors for promoting gastrointestinal health, it is essential to collect milk with consistently high growth factor activity. Therefore, we investigated the factors affecting growth factor activity in goat milk. Among the 5 breeds of dairy goats tested, milk from Nubian goats had the highest growth factor activity. Tight junction leakage induced by a 24-h milking interval did not increase growth factor activity in the milk. Milk collected from pregnant does had a significantly higher growth factor activity than milk collected postpartum. Growth factor activity decreased during the first 8 wk of lactation, fluctuated thereafter, and then increased dramatically after natural mating. During wk 1 to 8, growth factor activity was inversely correlated with milk yield and week of lactation. No correlation was observed during wk 9 to 29. After natural mating of the goats, the growth factor activity in the milk correlated significantly with somatic cell count and conductivity (a measure of membrane permeability), and correlated inversely with milk yield. Based on the above data, goat milk with higher growth factor activity could be selectively collected from Nubian pregnant does. PMID- 16702259 TI - The linear relationship between the proportion of fresh grass in the cow diet, milk fatty acid composition, and butter properties. AB - Fresh grass in the cow diet improves the rheological and nutritional properties of butter. However, the relationship between the proportion of fresh grass in the diet and these properties is still unknown. The objective of the study was to determine the relationship between the proportion of fresh grass in the diet and the properties of milk and butter. Four groups of 2 cows were fed 4 isoenergetic diets characterized by increasing amounts of fresh grass (0, 30, 60, and 100% dry matter of forage) according to a Youden square design. Energy levels were similar among all diets. Thus, no effect of mobilization was observed and the results were only due to the proportion of fresh grass in the diet. Milk yield linearly increased with the proportion of fresh grass in the diet (+0.21 kg/d per 10% of grass). Fat yield remained unchanged. Thus, by effect of dilution, increasing the proportion of fresh grass in the diet induced a linear decrease in fat content. Milk fat globule size decreased by 0.29 mum when the proportion of grass reached 30% in the diet. Increasing the proportion of fresh grass in the diet induced a linear increase in unsaturated fatty acids percentages at the expense of saturated fatty acids. Relationships were +0.38, +0.12, +0.05 and -0.69 points/10% of fresh grass in the diet for C18:1 trans-11, C18:2 cis-9,trans-11, C18:3n-3, and C16:0, respectively. These modifications in fatty acid composition, and in particular in the spreadability index, C16:0/C18:1, were responsible for linear decreases in final melting temperature and solid fat content in butter fat, perceived in sensory analysis by a linear decrease in firmness in mouth. The nutritional value of butter was also linearly improved by the proportion of fresh grass in the diet by halving the atherogenicity index. PMID- 16702260 TI - Oxidative stability of milk influenced by fatty acids, antioxidants, and copper derived from feed. AB - Differences in the oxidative stability of milk from cows fed grass-clover silage or hay were examined in relation to fatty acid composition and contents of antioxidants and copper in the milk. The oxidation processes were induced by exposing the milk to fluorescent light. Protein oxidation was measured as an accumulation of dityrosine, whereas lipid oxidation was measured as an accumulation of lipid hydroperoxides as the primary oxidation product, and as the secondary oxidation products, pentanal, hexanal, and heptanal. No differences were found in the protein oxidation of the 2 types of milk measured as accumulation of dityrosine, but there was an increased accumulation of lipid hydroperoxides and hexanal in milk from cows fed grass-clover silage, compared with milk from cows fed hay. The higher degree of lipid oxidation in milk from cows fed grass-clover silage could not be explained from the concentration of alpha-tocopherol, carotenoids, uric acid, and copper in the milk. However, it was thought to be highly influenced by the significantly higher concentration of linolenic acid present in milk from cows fed grass-clover silage. A larger part of alpha-tocopherol and beta-carotene was transferred from the feed to the milk when cows were fed grass-clover silage than when cows were fed hay as roughage. The significantly higher concentration of polyunsaturated fatty acids in milk from cows fed grass-clover silage may be important for the better transfer of alpha-tocopherol from the feed to the milk. Other circumstances, as the different conditions in the rumen may also play a role, due to the different types of roughages and their digestibility, or be related to the mechanisms during milk production for the higher yielding cows fed grass-clover silage. PMID- 16702261 TI - Precalving effects on metabolic responses and postpartum anestrus in grazing primiparous dairy cows. AB - The effect of increased access to pasture feeding during the last 6 wk of gestation on metabolic responses and postpartum anestrous interval was investigated. Heifers with a body condition score (BCS) of 5.0 (BC5+FF; on a 1-to 10 scale, US = 1.5 + 0.32 x New Zealand) were offered unrestricted pasture, and those with BCS 4.0 were fed either pasture unrestricted (BC4+FF) or restricted (BC4+RES) for the last 6 wk of gestation. After calving, all groups were offered unrestricted pasture. Mean BCS at calving for BC5+FF, BC4+FF, and BC4+RES were 4.7 +/- 0.1, 4.3 +/- 0.1, and 3.5 +/- 0.1, respectively. At 35 d postpartum, LH pulse frequency was lower in BC4+RES than in BC4+FF and BC5+FF, which were similar. At 77 d after calving, 8% of BC4+RES cows had ovulated compared with 75% of BC4+FF and 69% of BC5+FF cows. Metabolic hormonal differences between BC4+FF and BC4+RES were not reflected in the differences between BC4+FF and BC5+FF for LH pulse frequency or ovulation. Unrestricted access to pasture during the final 6 wk of gestation for BC4 heifers reduced the risk of prolonged postpartum anestrus. Systemic factors, tissue sensitivity, and critical developmental set points are probably involved in the integrated control of ovulation by body condition. PMID- 16702262 TI - Effects of housing, management, and health of dairy heifers on first-lactation udder health in southwest Sweden. AB - Data on health, management, and housing from birth to first calving were collected for 2,126 heifers on semi-monthly visits made by project veterinarians to 107 dairy herds from southwest Sweden. Additional data were obtained from the official milk- and health-recording program. Factors associated with incidence of veterinarian-treated clinical mastitis (VTCM) in the period 7 d before (d -7) to 30 d after first calving and of elevated cow composite somatic cell count (SCC, > or =200,000 cells/mL) at first test milking after first calving, respectively, were investigated using a 2-level (animal and herd) logistic regression analysis after initial screening by univariate analyses. The incidence risk of VTCM during the complete first lactation (305 d) was 10.8%. Ten percent of the diseased animals had more than 1 case of VTCM and 51% of total cases occurred from -7 to 30 d postcalving. The incidence rate of VTCM during the complete first 305-d lactation was 1.13 cases per 100 cow-mo. In total, 18.1% of the animals had elevated SCC at first test milking (mean 21 d) after calving. Veterinarian treated clinical mastitis at -7 to 30 d postcalving was associated with higher overall incidence of mastitis in the herd and with reproductive disorders (i.e., retained placenta, endometritis, pyometra, dystocia, or twin birth). The risk of elevated SCC increased with increasing percentage of cows in the herd that, some time during the year, had had an increased udder disease score (chronically increased SCC). Other factors associated with increased risk of elevated SCC were increasing amounts of concentrates fed to 11- to 16-mo-old heifers, moving to confined housing the day of calving instead of earlier, and use of restraint measures at milking. In addition, growth rate from birth to weaning, and several feed-related variables (e.g., amount of concentrates and type of roughage given) were associated with VTCM at -7 to 30 d post-calving or elevated SCC at first test milking in the univariate analyses. PMID- 16702263 TI - Technical note: Effects of adding shade and fans to a feedbunk sprinkler system for preparturient cows on health and performance. AB - The purpose of this study was to examine the effects of adding shades and fans to a feedbunk-mounted sprinkler system on preparturient Holstein cows during summer heat stress. Outcome variables included postpartum milk production, changes in body condition score, changes in serum concentrations of nonesterified fatty acids (NEFA), and incidence of postparturient disorders. Four hundred and seventy five prepartum multiparous cows, 250 to 257 d pregnant, were randomly allocated to 2 study pens. Treatments consisted of sprinklers over the feed bunk (n = 236); and sprinklers, fans, and shades over the feed bunk (n = 239). Data were used from cows spending a minimum of 14 d in their assigned pen. After parturition, all cows were housed and managed under identical conditions. Data recorders in each pen recorded environmental temperature and humidity every 30 min. Body condition scores were taken at study enrollment, parturition, and 60 d in lactation. Following parturition, the presence of retained placenta, metritis, milk fever, and displaced abomasum were recorded for the length of the study. Milk production was measured using twice-monthly Dairy Herd Improvement Association tests for the first 60 d in lactation. Blood was sampled twice weekly in 98 cows and analyzed for serum NEFA during the last 3 wk before parturition. Cows spent approximately 28 d in their respective treatments. Average daily environmental temperature (+/- SD) in the sprinkler only treatment was 26.4 +/- 7.2 vs. 25.1 +/- 8.6 degrees C in the shade, fans, and sprinkler treatment during the length of the trial. There was no difference in body condition score changes, incidence of postparturient disorders, or serum NEFA concentrations. There was a significant difference in total 60-d milk production, and an economic benefit over the preexisting cooling system. PMID- 16702264 TI - Longer milking intervals alter mammary epithelial permeability and the Udder's ability to extract nutrients. AB - Increasing the milking interval decreases milk yield and modifies milk composition. To gain a clearer understanding of the regulation of milk yield and composition, a study was conducted to establish the response curves of nutrient extraction by the mammary gland and mammary epithelial permeability in response to increasing milking intervals. Four multiparous lactating dairy cows were milked at 8-, 12-, 16-, or 24-h intervals over a period of 7 d using a Latin square design. Between the 8- and 24-h milking intervals, milk yield and milk protein levels fell curvilinearly from 38.2 to 29.2 kg/d and from 1,086 to 827 g/d, respectively. Milk fat yield decreased linearly from 1,475 to 1,235 g/d. Indicators of the opening of tight junctions increased linearly with increasing milking intervals: milk BSA increased from 148 to 207 mg/L and plasma lactose increased from 22.9 to 32.0 mg/L. The mammary gland's ability to extract nutrients decreased with increasing milking intervals. Extraction rates of glucose, beta-hydroxybutyrate, and total glycerol decreased significantly (from 27.2 to 23.3%, from 42.3 to 34.4%, from 36.6 to 30.8% between 8- and 24-h milking intervals, respectively), and not significantly for alpha-amino nitrogen (from 23.2 to 20.0%). The extraction rate of acetate remained constant. Moreover, the extraction of milk fat precursors appeared to be less regulated than those of the precursors of milk protein and lactose, which could partly explain why milk yield and milk protein yield decreased more than milk fat yield. The arteriovenous differences of beta-hydroxybutyrate and total glycerol remained constant, whereas those of glucose decreased significantly from 0.98 to 0.87 +/- 0.05 mmol/L and not significantly from 0.74 to 0.64 +/- 0.12 mmol/L for alpha-amino nitrogen. As a result, the mammary gland's ability to extract nutrients appears to be downregulated explaining partly the decrease in daily milk yield observed in response to increased milking intervals. PMID- 16702265 TI - Yields and persistency of lactation in Friesian and Jersey cows milked once daily. AB - Effects of milking cows once daily throughout lactation at high stocking rates (17% more cows/ha than for those milked twice daily) were studied in 2 Friesian and 2 Jersey herds during 3 lactations. Cows were allocated to 2 herds within breed and were milked either once or twice daily, based on age, genetic merit, and previous performance. Cows remained in their original herd and were milked at the same milking frequency during all lactations. Culled cows (20% per year) were replaced by 2-yr-old heifers. Yields of milk, lactose, protein, and fat were measured every 2 wk by commercial herd test. Cubic splines (5 knots) were used to approximate the lactation curve for each cow-yr to provide estimates of performance for each day of lactation. Yields of milk were greater for Friesian and Jersey cows milked twice daily (4,751 +/- 89 and 3,067 +/- 81 kg/cow) than for cows milked once daily (3,329 +/- 80 and 2,431 +/- 75 kg/cow), respectively. Cows milked once daily had lesser total and peak yields of milk, lactose, protein, and fat than cows milked twice daily. Friesians had greater total and peak yields than Jerseys. Peak production for all milk components occurred earlier in lactation for cows milked once daily than twice daily (d 24 to 39 vs. 32 to 44). Three measures of persistency of lactation were considered for each cow with 2 measures (Pers1 and Pers2) indicating that cows milked twice daily had better persistency than those milked once daily. Ranking of herds in persistency tended to match the ranking based on total yields. Measures of persistency (Pers1 and Pers2) were positively related to total yield in the Jerseys milked once daily and negatively related to peak yield in the Friesians. The third persistency measure (Pers3) ranked once-daily Jerseys first and twice-daily Friesians last, and was negatively correlated with total yield in the Friesian herds and negatively correlated with peak yield in all herds. For most performance measures, cows milked twice daily had better total yields and persistency than cows milked once daily. PMID- 16702266 TI - Contribution of somatic cell-associated activation of plasminogen to caseinolysis within the goat mammary gland. AB - Functional regression of the mammary gland is partly reflected by proteolysis of milk protein and tissue protein. The involvement of the plasminogen activation system in degradation of milk protein and mammary tissue damage has been demonstrated under inflammatory conditions. In this study, mammary secretion from 23 dairy goats primarily grouped as lactation (milking twice daily) or involution (milking once daily or less) was used to determine the ratio of gravity precipitated casein to total milk protein (casein ratio) as an index of caseinolysis, and activities of components of plasminogen activation system as well as their expressions on somatic cells. Based on the casein ratio, lactation goats were subcategorized as very active (71.8 +/- 1.0%) or less active (29.9 +/- 1.0%) in mammary function; involution goats were subcategorized as gradual (21.7 +/- 1.0%) or acute (5.9 +/- 0.2%) involution. This result suggests that caseinolysis occurred during regular lactation as well as during involution. On the other hand, activities of components of the plasminogen activation system in mammary secretion were increased along with the decreasing casein ratio, in contrast to the similar activities of their counterparts in circulation throughout various mammary statuses. Correlation analysis between casein ratio and activities of plasminogen activation system of goat milk indicated a significant negative relationship for plasmin (r = -0.64), plasminogen (r = 0.69), and urokinase-type plasminogen activator (uPA; r = -0.78) during involution but not during lactation. As for the cellular components of plasminogen activation system, there was an increase in immunoreactivity on somatic cells toward both monoclonal antibodies of human uPA and human uPA receptor under involution conditions suggesting their upregulation relative to lactation condition. Collectively, these results suggest that plasminogen activation system within the mammary gland differentially contribute to milk caseinolysis along the various stages of goat lactation. Meanwhile, a somatic cell-mediated local elevation of plasmin activity may be committed to extensive caseinolysis during involution. PMID- 16702267 TI - Antimicrobial susceptibility of Salmonella from organic and conventional dairy farms. AB - The objective of this study was to compare antimicrobial susceptibility of Salmonella isolated from conventional and organic dairy farms in the Midwest and Northeast United States. Environmental and fecal samples were collected from organic (n = 26) and conventional (n = 69) farms in Michigan, Minnesota, New York, and Wisconsin every 2 mo from August 2000 to October 2001. Salmonella isolates (n = 1,243) were tested using a broth microdilution method for susceptibility to amoxicillin-clavulanic acid, ampicillin, ceftiofur, ceftriaxone, cephalothin, chloramphenicol, ciprofloxacin, gentamicin, kanamycin, nalidixic acid, streptomycin, sulfamethoxazole, tetracycline, and trimethoprim sulfamethoxazole. Herd-level logistic regression and logistic proportional hazards multivariable models were used to examine the association between farm management type and susceptibility to antimicrobial agents. For most antimicrobial agents tested, susceptibility of Salmonella isolates was similar on organic and conventional herds when controlling for herd size and state. Conventional farms were more likely to have at least one Salmonella isolate resistant to streptomycin using logistic regression (odds ratio = 7.5; 95% confidence interval = 1.7-5.4). Conventional farms were more likely to have Salmonella isolates with greater resistance to streptomycin (odds ratio = 5.4; 95% confidence interval = 1.5-19.0) and sulfamethoxazole (odds ratio = 4.2; 95% confidence interval = 1.2-14.1) using logistic proportional hazards models. Although not statistically significant, conventional farms tended to be more likely to have at least one Salmonella isolate resistant to 5 or more antimicrobial agents when compared with organic farms. PMID- 16702268 TI - Dopamine antagonist alters serum cortisol and prolactin secretion in lactating Holstein cows. AB - The role of dopamine in regulating glucocorticoid and prolactin secretion was investigated in lactating Holstein cows by characterizing serum cortisol and prolactin responses to fluphenazine, a dopamine receptor antagonist. Twelve anovulatory cows received an intravenous bolus injection of either saline (n = 6) or 0.3 mg of fluphenazine/kg of body weight (n = 6) in wk 2 postpartum. Blood samples were collected every 30 min for 4 h before and 4 h after saline or fluphenazine injection. Serum progesterone concentration was 0.13 +/- 0.1 ng/mL and did not differ between groups. No difference in serum cortisol concentrations was detected between groups before treatments. Fluphenazine increased serum cortisol concentrations within 30 min after fluphenazine administration (>30 ng/mL) and concentrations remained elevated throughout the sampling period. Cortisol remained unchanged in saline-treated cows (<10 ng/mL). Prolactin concentrations also increased after fluphenazine administration (103.1 +/- 3.1 ng/mL), but were unaffected by saline (18 +/- 3.1 ng/mL). Prolactin concentrations remained elevated throughout the sampling period in fluphenazine treated cows. Our results indicated that a dopamine antagonist increased cortisol, suggesting that endogenous dopamine, at least in part, regulates cortisol and prolactin secretion. These effects are regulated through dopamine receptors in anovulatory lactating dairy cows during the early postpartum period. PMID- 16702269 TI - Embryo survival from gossypol-fed heifers after transfer to lactating cows treated with human chorionic gonadotropin. AB - Objectives were to determine the effects of gossypol exposure during early embryo development on embryonic survival after transfer of frozen and thawed embryos to lactating dairy cows treated with human chorionic gonadotropin (hCG). Holstein cows (n = 269) were either treated or not treated with 3,300 IU of hCG on d 5 of the estrous cycle and received an embryo collected from heifers fed or not fed gossypol. Embryo donor heifers consumed either 0 or 12 g/d of free gossypol for 76 d prior to embryo collection, resulting in mean plasma gossypol concentrations of 0 and 7.38 microg/mL, respectively. Embryos were transferred on d 7 of the estrous cycle and pregnancy diagnosed 21 and 35 d later. Progesterone was analyzed in plasma collected on d 5 and 12 of the estrous cycle. Treatment with hCG increased the total luteal area on d 12 (818.0 vs. 461.1 mm2) because of increased number of corpora lutea (2.0 vs. 1.0) and increased area of the original corpora lutea (522.7 vs. 443.5 mm2). Plasma progesterone concentrations were similar between treatments on d 5, but increased by d 12 in hCG-treated cows (6.46 vs. 4.78 ng/ mL). Pregnancy rates on d 28 and 42 were not affected by hCG. However, after transfer into lactating cows, embryos collected from heifers not fed gossypol resulted in higher pregnancy rates at 28 d (33.3 vs. 23.1%) and 42 d (29.6 vs. 20.2%) of gestation compared with embryos collected from heifers fed gossypol. Our data suggest that the negative effects of gossypol on fertility are mediated by changes in embryo viability in spite of similar grade quality at transfer. PMID- 16702270 TI - Flooring in front of the feed bunk affects feeding behavior and use of freestalls by dairy cows. AB - In 2 experiments we assessed how preferences, time budgets, and feeding behavior of dairy cows change in response to flooring surfaces in front of the feed bunk. In Experiment 1, 12 nonlactating dairy cattle were individually housed with access to 2 standing platforms filled with either concrete or sawdust. In Experiment 2, 24 nonlactating dairy cattle were given access to either concrete or Animat rubber flooring in front of the feed bunk. In Experiment 1, cows preferred the sawdust to the concrete flooring. In both experiments, cows provided with a softer floor in front of the feed bunk spent more time standing near the feed bunk without eating (Experiment 1: 67 vs. 40 min/d on sawdust vs. concrete, respectively, SEM = 5.6 min/d; Experiment 2: 176 vs. 115 min/d on Animat vs. concrete, respectively, SEM = 20.5 min/d) compared with when they were kept on concrete. The increased time spent at the feed bunk was due to a combination of more frequent eating and standing bouts, indicating that cows were more willing to move on nonconcrete flooring. Total time spent eating was significantly greater on the softer floor in Experiment 2, but not in Experiment 1 (Exp. 1: 289 vs. 275 min/d on sawdust and concrete, respectively, SEM = 7.3 min/d; Exp. 2: 330 vs. 289 min/d on Animat and concrete, respectively, SEM = 15.4), although feed intake was increased on the sawdust treatment in Experiment 1. Cows spent significantly more time lying in the feed alley when the flooring was rubber (219 vs. 53 min/d on Animat and concrete, SEM = 53.6 min/d), perhaps because the lying area in Experiment 2 was inadequate. In conclusion, cows prefer to stand on softer flooring in front of the feed bunk, and are more willing to move on and spend more time standing in front of the feed bunk when provided with softer flooring. These results indicate that cows find softer flooring surfaces more comfortable to stand on than concrete, and highlight the importance of evaluating the comfort of the entire facility. PMID- 16702271 TI - Short communication: Effects of Lactobacillus helveticus-fermented milk on the differentiation of cultured normal human epidermal keratinocytes. AB - Effects of Lactobacillus helveticus-fermented milk whey on the differentiation of normal human epidermal keratinocytes were studied. Analysis using real-time reverse transcription-polymerase chain reaction revealed that addition of Lactobacillus helveticus-fermented milk whey to the culture medium enhanced mRNA expression of keratin 10, an early differentiation marker, as well as involucrin, a late differentiation marker. Whey of artificially acidified milk, prepared by the addition of dl-lactic acid to milk instead of fermentation, also promoted expression of both markers, but Lactobacillus helveticus-fermented milk whey was more effective in increasing expression of those markers. These results indicate that milk whey has the potential to induce multiple stages of keratinocyte differentiation and that fermentation with Lactobacillus helveticus increases that activity. Furthermore, we examined the expression of profilaggrin, which increases with epidermal terminal differentiation, and found that Lactobacillus helveticus-fermented milk whey enhanced expression of profilaggrin mRNA in a dose dependent manner. Expression also occurred to a greater extent than with artificially acidified milk whey or other whey samples prepared with several lactic acid bacterial species. Because the proteolytically processed form of profilaggrin, filaggrin, is very important for normal epidermal hydration and flexibility, our results indicate that Lactobacillus helveticus-fermented milk whey has the potential to enhance the production of filaggrin-related natural moisturizing factor, because of its effect on the induction of epidermal differentiation, and is expected to be a useful skin moisturizing agent. PMID- 16702272 TI - Short communication: Correlations between udder morphology, milk yield, and milking ability with different milking frequencies in dairy goats. AB - Tinerfen a breed goats were assigned to 2 experimental herds and milked once (n = 28) or twice (n = 24) daily to study correlations between udder morphology, milk yield, and milking ability during the middle stage of the first lactation. Pearson correlation coefficients were significantly higher between yield and measures of udder globulousness (udder volume, r = 0.79 and r = 0.59; perimeter of insertion of the udder, r = 0.47 and r = 0.37; distance between teats, r = 0.77 and r = 0.28, for goats milked once and twice daily, respectively) than for length parameters (cistern floor distance, r = 0.40 and r = -0.29; udder depth, r = -0.20 and r = 0.20). The globulousness of the udder was correlated with easier milking ability, as shown by milk fractioning (r = 0.49 to 0.70) and milk flow measures (r = 0.32 to 0.49). The results showed that the globulousness of the udder is more important than length measurements in assessing milk yield and milking ability. PMID- 16702273 TI - Exposure to long-day photoperiod prepubertally may increase milk yield in first lactation cows. AB - Exposure to a long-day photoperiod (LDPP) increases mammary and lean growth in heifers relative to a short day (SDPP). Whether these effects influence milk yield, however, is unknown. To test the hypothesis that pre-pubertal long-day exposure would increase milk production, we assigned prepubertal heifers to LDPP (16 h of light:8 h of dark; n = 16) or SDPP (8 h of light:16 h of dark; n = 16) until the onset of puberty. At puberty, heifers were commingled and housed under natural photoperiodic conditions and standard nutritional management for dairy heifers. Heifers were bred according to body weight (BW = 385 kg) and withers height (WH = 132 cm). At parturition, BW, WH, and hip height were evaluated. Analysis of Dairy Herd Improvement Association records was used to evaluate milk yield, milk components, and somatic cell scores. Relative to SDPP (n = 12), heifers on LDPP (n = 10) had greater WH (by 2.4 cm) and BW at calving (by 55 kg). Compared with heifers housed under SDPP, LDPP heifers tended to produce more milk (750 kg of 305-d projected fat-corrected milk) during the first lactation. Relative to SDPP, prepubertal heifers exposed to LDPP during the prepubertal growth phase were taller and heavier at parturition, and tended to produce more milk during their first lactation. PMID- 16702274 TI - Effects of milking on dairy cow gait. AB - We studied cows with (n = 6) and without (n = 26) sole ulcers before and after milking to explore how milking influences dairy cattle gait and how this differs for cows with hoof injuries. Video recordings of cows were digitized using motion analysis software to calculate stride variables for each hoof. Gait was scored using a 1-to-5 numerical rating system (1 = sound, 5 = severely lame) and a continuous 100-unit visual analog scale of gait attributes (back arch, head bob, tracking-up, and reluctance to bear weight). For cows with and without sole ulcers, differences in gait before and after milking were evident; after milking, all cows had significantly longer strides (123.3 vs. 133.5 +/- 2.0 cm, respectively), higher stride height (8.3 vs. 8.9 +/- 0.1 cm), shorter stride durations (1.49 vs. 1.41 +/- 0.03 s), walked faster (0.85 vs. 0.97 +/- 0.03 m/s), and had shorter periods of triple support (3 legs in ground contact; 80.0 vs. 71.7 +/- 2.0%). Tracking-up and reluctance to bear weight improved after milking (20 vs. 16 +/- 2; 20 vs. 15 +/- 1, respectively), but numerical rating scores and back arch did not. Cows with sole ulcers walked differently than cows without for all measures, except swing duration, both before and after milking. Interactions between hoof health and milking were found for swing duration and head bob but when tested separately, the only effect was that cows without sole ulcers had longer swing durations before milking (0.45 vs. 0.44 +/- 0.01 s, respectively). Gait differences were probably due to udder distention and motivation to return to the home pen. Our results suggest that the most suitable time to assess lameness is after milking when differences between cows with and without ulcers are most evident. PMID- 16702275 TI - Effects of prepartum intramammary antibiotic therapy on udder health, milk production, and reproductive performance in dairy heifers. AB - Preparturient heifers (n = 561) from 9 herds in 6 US states and 1 Canadian province were enrolled in a study to test the hypothesis that prepartum intramammary therapy would cure existing intramammary infections (IMI) and lead to increased milk production, reduced linear somatic cell count (LSCC), and improved reproductive performance. Mammary secretions were collected 10 to 21 d before expected calving from each quarter. Heifers were then assigned by identification number to receive intramammary therapy consisting of infusion of one tube per mammary quarter of a lactating cow commercial antibiotic preparation containing cephapirin or to a nontreated control group. Overall, 34.1% of mammary quarters were infected with a mastitis pathogen before parturition and 63.4% of heifers had at least one mammary quarter infected. The coagulase-negative staphylococci (CNS) caused the majority (74.8%) of prepartum IMI. Coagulase positive staphylococci, environmental streptococci, and coliforms accounted for 24.5% of prepartum infections. Treatment had a significant effect on the cure rate of infected mammary quarters. Mammary quarters that were infected prepartum and treated with antibiotics had a 59.5% efficacy of cure rate and the percentage reduction in heifers with IMI was 51.9. Control quarters had a spontaneous cure rate of 31.7%. Treatment did not significantly affect milk production or LSCC in the first 200 d of lactation; however, there was a significant treatment by herd interaction for milk production. Quarters cured of either CNS or major pathogens had a lower LSCC in the first 200 d of lactation. No significant effect on services per conception or days open between treatment and control groups was observed. This trial demonstrated that prepartum intramammary antibiotic therapy did reduce the number of heifer IMI postpartum. Milk production, LSCC, and reproductive performance during the first 200 d of the first lactation were not significantly affected by treatment. Given these results, use of prepartum intramammary antibiotic therapy in heifers as a universal strategy to increase milk production in first-lactation dairy cows may not be warranted. PMID- 16702276 TI - Effect of interval from timed artificial insemination to initiation of resynchronization of ovulation on fertility of lactating dairy cows. AB - To compare 2 strategies for systematically resynchronizing ovulation, lactating Holstein cows (n = 763) at various days in milk and prior artificial insemination services were assigned randomly at timed AI (TAI) to receive the first GnRH injection of Ovsynch 26 (D26) or 33 (D33) d after TAI to resynchronize ovulation (Resynch) in cows failing to conceive. Cows in the D26 treatment received GnRH 26 d after TAI and continued Resynch only when diagnosed not pregnant by using ultrasonography 33 d after TAI, whereas D33 cows initiated Resynch only when diagnosed not pregnant 33 d after TAI. Cows were classified based on the presence or absence of a corpus luteum (CL) at the not-pregnant diagnosis, and cows without a CL received an intravaginal progesterone-releasing insert during Resynch. When analyzed as a systematic strategy, pregnancy rate per AI (PR/AI) was greater for cows assigned to the D33 than the D26 Resynch treatment (39.4 vs. 28.6%). A treatment x parity interaction was detected for PR/AI after Resynch for nonpregnant cows having a CL in which primiparous cows had a greater PR/AI than multiparous cows when Resynch was initiated 33 d after the initial TAI, and primiparous and multiparous cows when Resynch was initiated 26 d after the initial TAI. Pregnancy loss for Resynch was 6.4% between 33 and 40 d, and 2.6% between 40 and 61 d after Resynch TAI. We concluded that delaying initiation of Resynch until 33 d after TAI increased PR/AI for primiparous cows. PMID- 16702277 TI - Heat treatment of bovine colostrum. I: effects of temperature on viscosity and immunoglobulin G level. AB - The objective of this study was to identify the critical temperature, at or below which heat-treatment of bovine colostrum would produce no significant changes in viscosity, IgG concentration, or Ig activity. Results of preliminary work, using a Rapid Visco Analyzer (RVA) to heat 50-mL aliquots from 6 unique batches of bovine colostrum at 59, 60, 61, 62, and 63 degrees C, suggested that colostrum could be heated to 60 degrees C for up to 120 min without changing viscosity or IgG concentration. This finding was confirmed by heating 50-mL aliquots from 30 unique batches of colostrum in an RVA for 120 min at 60 and 63 degrees C. Heating colostrum to 63 degrees C resulted in an estimated 34% decrease in IgG concentration and 33% increase in viscosity. However, there was no difference in IgG concentration between preheat-treated (73.4 +/- 26.5 mg/mL) and post-heat treated (74.5 +/- 24.3 mg/mL) samples after heating colostrum to 60 degrees C in an RVA for 120 min. Similarly, viscosity was unaffected after heating colostrum to 60 degrees C in an RVA for 120 min. High quality colostrum (> or =73.0 mg/mL) suffered greater losses of IgG and greater viscosity changes when heated to 63 degrees C than did moderate quality colostrum (<73.0 mg/mL). However, the effects of colostrum quality were minor if high quality colostrum was only heated to 60 degrees C. The results of a bovine viral diarrhea serum neutralization assay suggested that antibody activity was unchanged after heating colostrum to either 60 or 63 degrees C. However, these results were interpreted as being inconclusive due to a high proportion of missing results because of the congealing of many samples after heat treatment. The results of this study indicate that 50-mL volumes of bovine colostrum can be heat treated at 60 degrees C for up to 120 min in an RVA without affecting IgG concentration or viscosity. PMID- 16702278 TI - Pre-conception energy balance and secondary sex ratio--partial support for the Trivers-Willard hypothesis in dairy cows. AB - According to the Trivers-Willard hypothesis, maternal condition at or around conception affects the secondary sex ratio in mammals. However, there are little or no data available on indicators of maternal condition in dairy cows on the sex of the resultant offspring. A total of 76,607 body condition score (BCS; scale of 1 to 5) records and 76,611 body weight (BW) records from 3,209 lactations across 1,172 cows were extracted from a research database collated from one research herd between 1986 and 2004, inclusive. Exclusion of multiple births and cows with no information before calving (e.g., nulliparous animals) resulted in 2,029 records with BCS and BW observations from the previous calving, and 2,002 and 1,872 lactations with BCS and BW observations at conception and midgestation, respectively. Change in BCS and BW between calving and conception and between conception and midgestation was calculated per lactation. Generalized estimating equations were used to model the logit of the probability of a male calf, in which cow was included as a repeated effect with a first-order autoregressive correlation structure assumed among records within cow. Of the BCS variables investigated, there was a linear relationship between the logit of the probability of a male calf and BCS change between calving and conception, the rate of BCS change over this period (BCS divided by days in milk), and BCS at the calving event immediately before conception. The birth of a bull calf was 1.85 times more likely in cows that lost no BCS from calving to conception compared with cows that lost one BCS unit from calving to conception. This increase in odds was equivalent to a 14% unit increase in the probability of a male calf (from 54 to 68%). The amount of BW lost between calving and conception and the rate of loss affected the sex of the resultant offspring. Less BW loss or greater BW gain between calving and conception was associated with greater likelihood of a male calf. Results suggested a positive effect of pre-conception BCS and BW change on secondary sex ratio, agreeing with the Trivers-Willard hypothesis that females in good physiological condition are more likely to produce male offspring. PMID- 16702279 TI - Effects of continuous versus periodic milk availability on behavior and performance of dairy calves. AB - Dairy calves fed milk ad libitum engage in feeding activity throughout the day and night. However, providing calves with continuous access to milk can present practical problems with milk quality, particularly during warm weather. One way to maintain milk quality in ad libitum feeding regimens is to limit the time that milk is available, but no research to date has addressed how this might affect calves. The objectives of this study were to quantify the effects of continuous access to milk (24 h/d) vs. access during only part of the day on milk feeding behavior and performance. Twenty-eight female Holstein calves were randomly assigned to treatments at 5 +/- 3 d of age and monitored for 28 d. All calves had milk delivered through a teat. Treatments were 1) access to milk for 24 h/d; 2) access to milk for 2 feedings per day (each of 2 h), with water available through the teat the remainder of the day (4-h wet treatment); and, 3) access to milk for 2 feedings per day (each of 2 h), but with no water available through the teat (4 h dry treatment). Calves given 24-h access to milk consumed 11.2 +/- 0.5 kg/d on average. Milk intake tended to be lower for the 4-h wet and 4-h dry treatments, averaging 10.0 +/- 0.3 kg/d, with no difference between these 2 treatments in this or any other variable. Average daily gain over the course of the experiment did not vary with treatment and averaged 1.1 +/- 0.04 kg/d. Calves in the 24-h treatment spent more time on the teat than those calves on the 4-h treatments, but during the 4 h/d when milk was available to all calves, calves in the 4-h treatments spent more time on the teat. These results show that individually housed calves fed milk ad libitum for only 4 h/d compensate by changing their milk feeding behavior and are able to achieve similar weight gains to animals fed milk continuously. PMID- 16702280 TI - An evaluation of the accuracy and precision of a stand-alone submersible continuous ruminal pH measurement system. AB - The objectives of this study were 1) to develop and evaluate the accuracy and precision of a new stand-alone submersible continuous ruminal pH measurement system called the Lethbridge Research Centre ruminal pH measurement system (LRCpH; Experiment 1); 2) to establish the accuracy and precision of a well documented, previously used continuous indwelling ruminal pH system (CIpH) to ensure that the new system (LRCpH) was as accurate and precise as the previous system (CIpH; Experiment 2); and 3) to determine the required frequency for pH electrode standardization by comparing baseline millivolt readings of pH electrodes in pH buffers 4 and 7 after 0, 24, 48, and 72 h of ruminal incubation (Experiment 3). In Experiment 1, 6 pregnant Holstein heifers, 3 lactating, primiparous Holstein cows, and 2 Black Angus heifers were used. All experimental animals were fitted with permanent ruminal cannulas. In Experiment 2, the 3 cannulated, lactating, primiparous Holstein cows were used. In both experiments, ruminal pH was determined continuously using indwelling pH electrodes. Subsequently, mean pH values were then compared with ruminal pH values obtained using spot samples of ruminal fluid (MANpH) obtained at the same time. A correlation coefficient accounting for repeated measures was calculated and results were used to calculate the concordance correlation to examine the relationships between the LRCpH-derived values and MANpH, and the CIpH-derived values and MANpH. In Experiment 3, the 6 pregnant Holstein heifers were used along with 6 new submersible pH electrodes. In Experiments 1 and 2, the comparison of the LRCpH output (1- and 5-min averages) to MANpH had higher correlation coefficients after accounting for repeated measures (0.98 and 0.97 for 1- and 5-min averages, respectively) and concordance correlation coefficients (0.96 and 0.97 for 1- and 5-min averages, respectively) than the comparison of CIpH to MANpH (0.88 and 0.87, correlation coefficient and concordance correlation coefficient, respectively). The concordance correlation analysis indicated that the ruminal pH data for LRCpH (1- and 5-min averages) vs. MANpH had location shifts that were smaller than those of the CIpH vs. MANpH. However, the scale shift was similar between the LRCpH and the CIpH. The plotted data from both systems closely resembled the line y = x, indicating that both systems were accurate and precise. In Experiment 3, changes in baseline millivolt readings for pH readings after 24, 48, or 72 h of ruminal incubation were not significantly different than zero, indicating that daily standardization of new electrodes was not essential. Results from this study indicate that the LRCpH system can accurately and precisely measure ruminal pH; thus, it provides increased opportunity for researchers to measure ruminal pH and the occurrence of ruminal acidosis in unrestrained cattle. PMID- 16702281 TI - Prepartal plane of nutrition, regardless of dietary energy source, affects periparturient metabolism and dry matter intake in Holstein cows. AB - Previous research in our laboratory showed that dietary fat supplementation during the dry period was associated with decreased peripartum hepatic lipid accumulation. However, fat supplementation decreased dry matter (DM) intake and thereby confounded results. Consequently, 47 Holstein cows with body condition scores (BCS) < or = 3.5 at dry-off were used to determine whether source or amount of energy fed to dry cows was responsible for the decreased hepatic lipid content. Moderate grain- or fat-supplemented diets [1.50 Mcal of net energy for lactation (NE(L))/kg] were fed from dry-off (60 d before expected parturition) to calving at either ad libitum (160% of NE(L) requirement) or restricted (80% of NE(L) requirement) intakes. Postpartum, cows were fed a single lactation diet for ad libitum intake and performance was measured for 105 d. Prepartum intakes of DM and NE(L) were significantly lower for feed-restricted cows as designed. During the first 21 d postpartum, previously restricted cows had higher intakes of DM and NE(L). Body weights and BCS were lower prepartum for restricted cows but groups converged to similar nadirs postpartum. Restricted-fed cows had lower concentrations of glucose and insulin and increased concentrations of NEFA in plasma during the dry period. Peripartum NEFA rose markedly for all treatments but were higher postpartum for cows previously fed ad libitum. Plasma concentrations of NEFA and BHBA remained lower in cows restricted-during the dry period. Postpartum concentrations of total lipid and triglyceride in liver were lower in cows previously feed-restricted. Across dietary treatments, activity of carnitine palmitoyltransferase (CPT) in hepatic mitochondria was lowest at - 21 d, highest at 1 d, and decreased at 21 and 65 d relative to parturition. The activity of CPT at d 1 tended to be higher for previously feed-restricted cows; thereafter, CPT activity declined more rapidly than in cows fed ad libitum. Nutrient intake during the dry period had more pronounced effects on peripartal lipid metabolism and DMI than did composition of the prepartum diet. PMID- 16702282 TI - Intake and performance of lactating cows grazing diverse forage mixtures. AB - Twenty multiparous Holstein cows in midlactation grazed pastures of 4 forage mixtures in a 12-wk study repeated during 2 grazing seasons to determine if forage mixture complexity affected intake and productivity of lactating dairy cows. The forage mixtures were 1) orchardgrass plus white clover [2 species (SP)]; 2) orchardgrass, white clover, and chicory (3SP); 3) orchardgrass, tall fescue, perennial ryegrass, red clover, birdsfoot trefoil, and chicory (6SP); and 4) 6SP mixture plus white clover, alfalfa, and Kentucky bluegrass (9SP). Total herbage intake was similar among forage mixtures, averaging 12.0 kg/d across all forage mixtures and years. Milk production and composition were not affected by forage mixture or year, and averaged 34.6 kg/d, 3.4%, and 2.8% for milk production, milk fat percentage, and milk protein percentage, respectively. The conjugated linoleic acid content of milk fat was higher for cows that grazed the 3SP, 6SP, and 9SP mixtures than from cows that grazed the 2SP mixture (1.02 vs. 0.87 g of conjugated linoleic acid/100 g of fatty acids, respectively). Blood glucose, blood urea nitrogen, and nonesterified fatty acids were not affected by forage mixture and averaged 69.2 mg/dL, 13.4 mg/dL, and 277.5 muEq/L, respectively. The results of this study indicate that altering the forage mixture in pastures did not affect dry matter intake, milk production, or blood metabolite profiles of lactating cows. The use of complex mixtures of forages in grazing systems should not affect dairy cow performance. PMID- 16702283 TI - Fiber and lignin analysis in concentrate, forage, and feces: detergent versus enzymatic-chemical method. AB - Hemicelluloses, cellulose, and lignin contents of contrasting feeds, with emphasis on concentrate ingredients and complete concentrates, were analyzed using the Van Soest detergent procedure (analyzing neutral detergent fiber, acid detergent fiber, and acid detergent lignin) and the enzymatic-chemical procedure (analyzing cellulose, soluble and insoluble noncellulosic polysaccharides, and Klason lignin). Also, feces from cows fed concentrates differing in carbohydrate composition were analyzed by the 2 procedures. The correlation between acid detergent lignin and Klason lignin was significant, but not as close as the one between individual structural polysaccharides measured with the 2 procedures. The correlation between the results of the 2 procedures was highly significant for apparent cellulose digestibility, as were the correlations between digestibilities of hemicelluloses with total as well as with insoluble noncellulosic polysaccharides. The relationship between dietary lignin content and fiber digestibility was weak. The exclusion of a group of cows fed a concentrate with apple pulp, however, improved the respective correlations. Klason lignin correlated more closely with the measured fiber digestibility than acid detergent lignin. The study showed that results of the detergent method were comparable to those of the enzymatic-chemical method with cellulose, hemicelluloses, and their digestibilities. However, acid detergent lignin was much lower than Klason lignin. When the carbohydrate composition of concentrate varied widely, lignin was not suitable for the prediction of fiber digestibility. PMID- 16702284 TI - Responses to amino acid imbalances and deficiencies in lactating dairy cows. AB - Lactating cows were exposed to large amino acid imbalances and deficiencies by i.v. infusion to characterize responses in milk production and plasma concentrations of metabolites and hormones. Six cows in early lactation were fed a basal diet of 9% CP and infused continuously for 6 d with saline (negative control), 1.1 kg/d of a complete amino acid mix (positive control), or the equivalent mix lacking Met, Lys, His, or all 3 branched-chain amino acids. All cows received all treatments in 6 successive periods in a Latin square design. Infusion of the complete amino acid mix resulted in an increase in the plasma concentrations of several essential amino acids, insulin, and glucagon. Milk protein production was stimulated by 19%, which accounted for 10% of the infused amino acid. Plasma urea, acetate, and beta-hydroxybutyrate concentrations were increased. Compared with saline, the amino acid mixtures lacking Met, Lys, or His increased essential amino acids, glucose, insulin, and glucagon concentrations in plasma, and decreased growth hormone. Plasma concentration of the essential amino acid absent from the infusate fell 2-fold but milk protein yield remained within 12% of its basal value. Dry matter intakes were depressed 35% over the first 2 d of infusion of imbalanced mixtures but recovered thereafter. Milk fat yields were increased 258 and 320 g/d by mixtures devoid of Lys and His, respectively. Correction of a Met, Lys, or His deficiency did not affect hormone concentrations in plasma and milk protein yield increased 27% due entirely to increased concentration of the single amino acid in plasma. Although imbalance and deficiency generated similar amino acid profiles in plasma, it was concluded that endocrine responses to total amino acid supply during imbalance can override imperfections in the circulating amino acid profile to maintain milk protein yield at higher levels than expected from deficiency states. Both imbalance and deficiency were characterized by a low protein:fat ratio in milk. Infusion of a mix of amino acids lacking Val, Ile, and Leu, despite a decrease in plasma Leu to 58% of its basal value, increased milk protein and fat yields to the same extent as the complete amino acid mix. PMID- 16702286 TI - Feasibility of a linear scoring method of udder morphology for the selection scheme of Sardinian sheep. AB - Breeders are increasingly interested in improving the machine milkability of Sardinian dairy sheep by selection for udder morphology. Nine-point linear scales were developed to appraise teat placement, degree of suspension of the udder, udder depth, and degree of separation of the 2 halves. Repeatabilities within and across lactation were estimated on an experimental flock. Ewes were scored at least 3 times a year from 1999 to 2003. Within-lactation repeatabilities were greater than 0.70 for all linear traits, whereas across-lactation repeatabilities ranged from 0.59 to 0.66. The first linear combination of basic traits, obtained by principal component analysis, was highly correlated with teat placement, degree of suspension of the udder, and udder depth and showed an across-lactation repeatability of 0.76. These results indicate that an accurate evaluation of animals is possible by a single, early lifetime score. Genetic parameters of linear udder traits were estimated using a REML method applied to a sire model. Data were selected from 76,984 scores of Sardinian yearling ewes collected by 31 classifiers from 1999 to 2004. Two models were compared, one fitting the contemporary group effect as fixed and the other as random. Heritabilities of udder traits ranged from 0.19 to 0.31. The model with the random contemporary group effect produced slightly higher heritabilities and higher correlations between the sires' estimated breeding values and the daughters' average scores. As a whole, the genetic correlations between udder traits were favorable, indicating that selection for one trait will produce a positive evolution of the overall udder conformation. In particular, the degree of suspension of the udder was highly correlated with udder depth (0.82). Genetic correlations with milk yield were unfavorable but generally low, with the exception of udder depth ( 0.48). Genetic trends were estimated using an animal model. Only udder depth showed a negative constant genetic trend. Overall results indicated that genetic improvement of the udder morphology of Sardinian ewes is feasible, with major emphasis on teat placement and degree of udder suspension, traits showing the highest heritabilities and low unfavorable genetic correlations with milk yield. PMID- 16702285 TI - Eating rate and preference of different concentrate components for cattle. AB - Different feeds or combination of feeds were studied in 11 short-term experiments with the objective to identify concentrates that were especially desirable for cattle. Eating rate of different feeds was studied in 6 experiments using 10 heifers in a Latin square design with 2 blocks of 5 animals, 5 treatments (feeds), and 5 periods (days). Preference as shown by feed choice was studied in 5 experiments with 12 heifers. Paired comparisons of 4 different feeds (1 through 4) in the 6 possible combinations of 1 and 2, 1 and 3, 1 and 4, 2 and 3, 2 and 4, and 3 and 4 were performed. A control feed of ground barley was included in all experiments. In the eating rate and preference experiments, a total of 25 and 16 feeds, respectively, were studied. The categories of feeds studied were basic feeds, such as cereals, soybean meal, and rapeseed products, and feed mixtures based on ground barley with sweet additives or additives based on fat products. Pelleted concentrate mixtures were also evaluated. From the results obtained, the following feeds were identified as being among the most preferred feeds: pelleted feeds, heat-treated rapeseed meal, barley with 10% rapeseed fatty acid, barley with 10% palm oil, and barley with 10% glycerol, whereas ground palm kernel expeller was undesirable. A clear preference for pellets over ground barley was demonstrated, but no difference in preference was observed for the 3 different pellets that were compared. PMID- 16702287 TI - Analysis of inbreeding and its relationship with functional longevity in Canadian dairy cattle. AB - The aim of this study was to assess the level of inbreeding and its relationship to the functional survival of Canadian dairy breeds by using a Weibull proportional hazard model. Data consisted of records from 72,385 cows in 1,505 herds from 2,499 sires for Jerseys, 112,723 cows in 1,482 herds from 2,926 sires for Ayrshires, and 1,977,311 cows in 17,182 herds from 8,261 sires for Holsteins. Longevity was defined as the number of days from first calving to culling, death, or censoring. Inbreeding coefficients (F) were grouped into 7 classes (F = 0, 0 < F < 3.125, 3.125 < or = F < 6.25, 6.25 < or = F <12.5, 12.5 < or = F < 18.25, 18.25 < or = F < 25.0, and F > or = 25.0%). The statistical model included the effects of stage of lactation, season of production, the annual change in herd size, type of milk recording supervision, age at first calving, effects of milk, fat, and protein yields calculated as within herd-year-parity deviations, herd year-season of calving, inbreeding, and sire. The relative culling rate was calculated for animals in each class after accounting for the above-mentioned effects. A trend toward increased risk of culling among more inbred animals was observed for all breeds. Little difference in survival was observed for cows with 0 < F <12.5%. The relative risk ratios (relative to F = 0) for cows with inbreeding coefficients up to 12.5% were 1.19, 1.16, and 1.14 for Jersey, Ayrshire, and Holstein cows, respectively. Greater effects of inbreeding were seen, however, when F increased beyond 12.5%. PMID- 16702289 TI - Genetic relationship between first-lactation body energy and later-life udder health in dairy cattle. AB - Weekly body condition score (BCS) and live weight records were used to calculate energy content (EC) and cumulative effective energy balance (CEEB) for 508 Holstein-Friesian cows in their first lactation. Cows were raised on an experimental farm and had calved between 1991 and 2000. Energy content was an estimate of the actual energy level of a cow at any given stage of lactation, whereas CEEB was associated with the total body energy content as defined by accumulated weekly energy balance changes since the onset of lactation. Genetic evaluations were computed for the 3 body energy traits (BCS, EC, and CEEB) for each week of first lactation. Random regression models were used to assess the association between first-lactation weekly genetic evaluations for body energy and monthly test-day log-transformed SCC, clinical mastitis, and other udder problems in the first 3 lactations. There was a significant effect of at least one body energy trait at any stage of first lactation past wk 3 on SCC in the first 3 lactations. Maximum genetic correlation estimates were -0.18 (+/-0.04) between wk-16 BCS and SCC in the first 2 lactations, -0.18 (+/-0.04) between wk 11 EC and SCC in the first 2 lactations, and -0.17 (+/-0.07) between wk-6 CEEB and SCC in the first 2 lactations. The effect of body energy traits on clinical mastitis was, in general, nonsignificant; nevertheless, moderate genetic correlations were estimated, ranging from -0.05 (+/-0.07) to -0.25 (+/-0.15). The effect of body energy traits on udder problems other than mastitis was negligible in all cases. Results suggest that, amongst the traits studied here, BCS, EC, and CEEB in the first 3 to 4 mo of lactation 1 had the greatest genetic association with SCC and mastitis in first, second, and, to a lesser extent, third lactations. PMID- 16702288 TI - Technical note: Whole-genome amplification of DNA extracted from cattle semen samples. AB - The bovine genome sequence project and the discovery of many thousands of bovine single nucleotide polymorphisms has opened the door for large-scale genotyping studies to identify genes that contribute to economically important traits with relevance to the beef and dairy industries. Large amounts of DNA will be required for these research projects. This study reports the use of the whole-genome amplification (WGA) method to create an unlimited supply of DNA for use in genotyping studies and long-term storage for future gene discovery projects. Two commercial WGA kits (GenomiPhi, Amersham Biosciences, Sydney, Australia, and REPLI-g, Qiagen, Doncaster, Australia) were used to amplify DNA from straws of bull semen, resulting in an average of 7.2 and 67 microg of DNA per reaction, respectively. The comparison of 3.5 kb of sequences from the amplified and unamplified DNA indicated no detectable DNA differences. Similarly, gene marker analysis conducted on genomic DNA and DNA after WGA indicated no difference in marker amplification or clarity and accuracy of scoring for approximately 10,000 single nucleotide polymorphism markers when compared with WGA samples genotyped in duplicate. These results illustrate that WGA is a suitable method for the amplification and recovery of DNA from bull semen samples for routine genomic investigations. PMID- 16702290 TI - Short communication: Optimal random regression models for milk production in dairy cattle. AB - Legendre polynomials of orders 3 to 8 in random regression models (RRM) for first lactation milk production in Canadian Holsteins were compared statistically to determine the best model. Twenty-six RRM were compared using LP of order 5 for the phenotypic age-season groupings. Variance components of RRM were estimated using Bayesian estimation via Gibbs sampling. Several statistical criteria for model comparison were used including the total residual variance, the log likelihood function, Akaike's information criterion, the Bayesian information criterion, Bayes factors, an information-theoretic measure of model complexity, and the percentage relative reduction in complexity. The residual variance always picks the model with the most parameters. The log likelihood and information theoretic measure picked the model with order 5 for additive genetic effects and order 7 for permanent environmental effects. The currently used model in Canada (order 5 for both additive and permanent environmental effects) was not the best for any single criterion, but was optimal when considering all criteria. PMID- 16702291 TI - Genetic associations between clinical mastitis and somatic cell score in early first-lactation cows. AB - The objectives of this study were to examine genetic associations between clinical mastitis and somatic cell score (SCS) in early first-lactation cows, to estimate genetic correlations between SCS of cows with and without clinical mastitis, and to compare genetic evaluations of sires based on SCS or clinical mastitis. Clinical mastitis records from 15 d before to 30 d after calving and first test-day SCS records (from 6 to 30 d after calving) from 499,878 first lactation daughters of 2,043 sires were analyzed. Results from a bivariate linear sire model analysis of SCS in cows with and without clinical mastitis suggest that SCS is a heterogeneous trait. Heritability of SCS was 0.03 for mastitic cows and 0.08 for healthy cows, and the genetic correlation between the 2 traits was 0.78. The difference in rank between sire evaluations based on SCS of cows with and without clinical mastitis varied from -994 to 1,125, with mean 0. A bivariate analysis with a threshold-liability model for clinical mastitis and a linear Gaussian model for SCS indicated that heritability of liability to clinical mastitis is at least as large as that of SCS in early lactation. The mean (standard deviation) of the posterior distribution of heritability was 0.085 (0.006) for liability to clinical mastitis and 0.070 (0.003) for SCS. The posterior mean (standard deviation) of the genetic correlation between liability to clinical mastitis and SCS was 0.62 (0.03). A comparison of sire evaluations showed that genetic evaluation based on SCS was not able to identify the best sires for liability to clinical mastitis. The association between sire posterior means for liability to clinical mastitis and sire predicted transmitting ability for SCS was far from perfect. PMID- 16702292 TI - Multitrait quantitative trait Loci mapping for milk production traits in danish Holstein cattle. AB - The aims of this study were (1) to confirm previously identified quantitative trait loci (QTL) on bovine chromosomes 6, 11, 14, and 23 in the Danish Holstein cattle population, (2) to assess the pleiotropic nature of each QTL on milk production traits by building multitrait and multi-QTL models, and (3) to include pedigree information on nongenotyped individuals to improve the estimation of genetic parameters underlying the random QTL model. Nineteen grandsire families were analyzed by single-trait (ST) and multitrait (MT) QTL mapping methods. The variance component-based QTL mapping model was implemented via restricted maximum likelihood (REML) to estimate QTL position and parameters. Segregation of the previously identified QTL was confirmed on bovine chromosomes 6, 11, and 14, but not on 23. A highly significant (1% chromosome-wise level) QTL was found on chromosome 6, between 37 and 73 cM. This QTL had a strong effect on protein percentage (PP) and fat percentage (FP) according to ST analyses, and effects on PP, FP, milk yield (MY), fat yield (FY), and protein yield (PY) in MT analyses. A QTL affecting PP was detected on chromosome 11 (at 70 cM) using ST analysis. The MT analysis revealed a second QTL (at 67 cM) approaching significance with an effect on MY. The ST analysis identified a QTL for MY and FP on chromosome 14, between 10 and 24 cM. The extended pedigree (nongenotyped animals) was included to estimate genetic parameters underlying the random QTL model; that is, additive polygenic and QTL variances. In general, the estimates of the QTL variance components were smaller but more precise when the extended pedigree was considered in the analysis. PMID- 16702293 TI - Inbreeding depression for global and partial economic indexes, production, type, and functional traits. AB - The objective of this research was to examine the effects of inbreeding in the population of Holstein cattle in the Walloon region of Belgium. The effects of inbreeding on the global economic index and its components were studied by using data from the genetic evaluations of February 2004 for production, somatic cell score (SCS), computed from somatic cell counts and type. Inbreeding coefficients for 956,516 animals were computed using a method that allows assigning an inbreeding coefficient to individuals without known parents. These coefficients were equal to the mean inbreeding coefficient of contemporary individuals with known parents. The significance of inbreeding effects on the different evaluated traits and on the different indexes were tested using a t-test comparing estimated standard errors and effects. The inbreeding effect was significantly different from zero for the vast majority of evaluated traits and for all of the indexes. Inbreeding had the greatest deleterious effects on production traits. Inbreeding decreased yield of milk, fat, and protein during a lactation by 19.68, 0.96, and 0.69 kg, respectively, per each 1% increase in inbreeding. The regression coefficient of SCS per 1% increase in inbreeding was +0.005 SCS units. The inbreeding depression was thus relatively low for SCS, but inbred animals had higher SCS than non-inbred animals, indicating that inbred animals would be slightly more sensitive to mastitis than non-inbred animals. Estimates of inbreeding effects on evaluated type traits per 1% increase were small. The most strongly affected type traits were chest width, rear leg, and overall development on a standardized scale. For several type traits, particularly traits linked to the udder, the estimates suggested a favorable effect of inbreeding. The global economic index was depressed by around 6.13 euro of lifetime profit per 1% increase in inbreeding for the Holstein animals in the Walloon region of Belgium. PMID- 16702294 TI - Validation of feed and manure data collected on Wisconsin dairy farms. AB - An on-farm study of 54 representative Wisconsin dairy farms was conducted to evaluate the influence of biophysical and socioeconomic factors on overall feed, fertilizer, and manure nutrient use. This report validates 1) how well data on cow diets, feed analyses, and milk production reflected established feed-milk manure relationships; and 2) how well farmer-recorded data on manure land application reflected literature values of manure N and P excretion, collection, and loss. Calculated feed N and P use efficiencies (18 to 33% and 18 to 35%, respectively) fell within ranges expected for dairy farms. This suggested that our on-farm methods of data collection provided reliable information on relationships among feed N and P intake, secretions in milk, and excretion in manure. On stanchion farms, there were no differences between farmer estimates (kg/farm) of manure P collected (1,140) and land-applied (1,210) and what would be calculated from the literature (1,340). On freestall farms, there were no differences in amounts (kg/farm) of manure P collected (2,889), land-applied (2,350), or literature estimates (2,675). Manure P applications (kg/ha) to tilled cropland would be similar using either farmer estimates of manure collected and land-applied, or literature estimates. The data provided a snapshot of Wisconsin industry practices, as well as information on the range of feed and manure management practices on individual dairy farms. Improvements to data collection methods would require increased skill and training of both farmers and those responsible for assisting farmers in on-farm data collection and analyses. PMID- 16702295 TI - Efficacy of two barrier iodine teat dips under natural exposure conditions. AB - The efficacy of 2 iodine barrier teat dips was compared with a conventional iodine postmilking teat dip. The products were evaluated using a natural exposure trial on a 250-cow dairy based on the National Mastitis Council guidelines. Bacteriological samples were taken every 2 wk over 6 mo. All dips contained 1.0% available iodine and high levels of free iodine for germicidal efficacy. Barrier 1 was an experimental dip containing 14 to 20 ppm of free iodine, barrier 2 contained 8 to 14 ppm of free iodine, and the nonbarrier control contained 12 to 16 ppm of free iodine. The average free iodine for barrier 1 and barrier 2 combined was 14 ppm, which was equal to the average free iodine for the positive control. A 21% reduction of new intramammary infections (IMI) was observed for the 2 barrier dips combined when compared with the nonbarrier control. A significant reduction of 38% was observed for the clinical infection rate for barrier 1 when compared with barrier 2; however, barrier 1 did not significantly reduce the subclinical IMI when compared with barrier 2. Barrier 1 significantly reduced the combined clinical and subclinical IMI by 24% when compared with the positive control. The skin condition showed slight variation among the dips over the course of the trial, but no difference was observed at the end of the trial. Barrier 1, with the highest concentration of free iodine, gave the best efficacy results of all 3 dips. PMID- 16702296 TI - Dairy cattle culling patterns, explanations, and implications. AB - Culling patterns in the Upper Midwest and Northeast regions were examined from Dairy Herd Improvement records from 1993 through 1999. There were 7,087,699 individual cow lactation observations of which 1,458,936 were complete. A probit regression model was used to determine how individual cow and herd characteristics affected the likelihood of a cow being culled. The model predicted whether individual cows were culled each month. With a combination of observable cow and herd characteristics, as well as variables to capture state, year, and farm effects, the model was able to explain, with a 79.5 and 79.9% accuracy rate, individual cow cull decisions in the Upper Midwest and Northeast regions, respectively. Summer (- 11.5% in the Upper Midwest; - 6.4% in the Northeast) and fall (- 18.7% in the Upper Midwest; - 7.9% in the Northeast) calving vs. spring calving, higher than average milk production (- 1.7% per hundredweight in the Upper Midwest; - 0.5% in the Northeast), higher than average protein content (- 0.2% per additional percentage milk protein in the Upper Midwest; - 0.1% in the Northeast), higher milk production persistency (- 2.1% per additional percent persistent in the Upper Midwest; - 1.8% in the Northeast), and expansion (during the early years following the expansion) were associated with a reduced likelihood of a cow being culled. Lower than average fat content (0.04% per additional percentage butterfat in the Upper Midwest; 0.02% in the Northeast), and higher than average somatic cell count (8.8% for each unit increase in somatic cell count score in the Upper Midwest; 7.8% in the Northeast) were associated with an increased likelihood of a cow being culled. The study results are useful in describing patterns of culling and relating them to cow, herd, geographic, and time variables and can act as a benchmark for producers. PMID- 16702297 TI - Opinions and practices of wisconsin dairy producers about biosecurity and animal well-being. AB - The objective of this study was to characterize opinions and practices of Wisconsin dairy producers about biosecurity and animal wellbeing. Wisconsin dairy producers were surveyed using a mailed questionnaire and responder herds were categorized based on the number of lactating cows: very small herds (< or =50 lactating cows; n = 279); small herds (51 to 100 lactating cows; n = 202); medium herds (101 to 200 lactating cows; n = 42); and large herds (>200 lactating cows; n = 37). Producers from large herds adopted more biosecurity practices than those from small herds, but biosecurity risks were common. Almost half of the responders indicated that they purchased cattle, but few (49.4%) performed diagnostic testing of those cattle. The frequency of diagnostic testing and examination of purchased cattle increased with herd size. Producers generally (80%) believed that they used the "right amount" of antibiotics, but the use of written treatment protocols increased with herd size. Producers from large and medium herds reported much higher usage of computerized (65.7 and 17.5%, respectively) and paper records (42.9 and 22.5%, respectively) compared with producers from smaller herds. Almost all (92.6%) believed that Johne's disease was an important issue for the dairy industry, but only 9% had enrolled in the official Wisconsin control program. Most producers (88.6%) believed that dehorning caused at least a small amount of pain, but the majority (81%) did not use local anesthetics. Producers minimized risks with which they were most familiar. Drinking raw milk was not considered a human health risk by almost half the responders, whereas bovine spongiform encephalopathy was considered "no risk" to only 37%. Raw milk was consumed by more than 60%, but regular consumption of raw milk decreased from 47.7% (very small herds) to 24.3% (large herds); perception of the risk of raw milk increased from 46.2% (very small herds) to 56.8% (large herds) with herd size. Larger farms had more knowledge of personal health risks related to zoonotic pathogens. Overall, most management practices were associated with herd size, but many beliefs regarding important dairy farm issues were consistent. PMID- 16702298 TI - Changes in terpene content in milk from pasture-fed cows. AB - Changes of terpene content in milk from cows grazing natural diversified upland pasture were examined in this observational trial. A homogeneous plot divided into 2 subplots was used from May 31 to July 1, 2003 (first growth) and again from October 1 to October 7, 2003 (vegetative regrowth). Each subplot was grazed by 6 dairy cows in 2 ways: strip grazing (SG), with new allocations of pasture strips at 2-d intervals, and paddock grazing (PG). The PG subplot was divided into 3 paddocks and the cows were moved to a new paddock on June 13 and June 24, 2003. Milk from the 6 cows was collected twice a week, pooled, and used for terpene analyses by dynamic headspace gas chromatography-mass spectrometry system. Twenty mono- and 23 sesquiterpenes desorbing from the milk fat were separated. The most abundant monoterpenes were beta-pinene, alpha-pinene, gamma terpinene, limonene, alpha-tujene, terpinolene, and alpha-phellandrene. The most abundant sesquiterpenes were beta-caryophyllene, alpha-copaene, beta-cedrene, transmuurola-4-(14)-5-diene, beta-bisabolene, and delta-cadinene. Both mono- and sesquiterpenes in SG milk increased across time with an 8-fold increase in total terpenes in milk from the beginning to the end of June. In parallel, dicotyledons, including the main terpene-rich plants, increased from 17 to 31% of total biomass of the vegetation and the development of Dactylis glomerata progressed from boot to ripening stage. The terpenes in PG milk were equivalent to those in SG milk for the first paddock at the beginning of June and remained constant or doubled for the sum of mono- and sesquiterpenes, respectively. The lower variability of the PG milk terpene content could be related to the opportunity that PG cows had to choose ingested herbage, whereas the SG cows had limited choice within the smaller allocated pasture strips. Milk from cows grazing regrowth pastures in October contained low levels of terpenes, and values were similar for PG and SG milks. Change in the sum of monoterpenes is representative of changes for most monoterpenes (except for limonene) and, conversely, the change in the sum of sesquiterpenes mainly reflected changes in beta-caryophyllene and alpha-copaene. In addition to effects of botanical composition of pasture, it appears that terpene content in milk may vary according to factors linked to grazing management that need more intensive study. Nevertheless, current results raise questions about the precision of terpenes as feed tracers. PMID- 16702299 TI - Utility of near-infrared reflectance spectroscopy to predict nutrient composition and in vitro digestibility of total mixed rations. AB - Total mixed ration (TMR) samples (n = 110) were analyzed for dry matter (DM), crude protein (CP), soluble CP, neutral detergent fiber (NDF), NDF CP, starch, ash, fat, total ethanol-soluble carbohydrate, and nonfiber carbohydrate (NFC). Rapidly and slowly degraded and undegraded in situ CP fractions and in vitro DM, organic matter, and NDF digestibility were determined on each TMR. The TMR were scanned using near-infrared reflectance spectroscopy (NIRS); spectra were retained with NIRS calibration and cross-validation statistics were determined using partial least squares regression methods. The CP, NDF, starch, in vitro DM, and in vitro indigestible NDF contents of TMR were predicted by NIRS with good degrees (R2 >0.85) of accuracy with proportionally low standard errors of prediction. Moderate utility of NIRS to predict the NFC (R2 = 0.83) and fat content (R2 = 0.81) of TMR was observed. Rapidly, slowly, and undegraded in situ CP fractions in TMR were not well predicted by NIRS. Similarly, soluble CP, NDF CP, total ethanol-soluble carbohydrate, and in vitro NDF digestibility (% of NDF) were not well predicted by NIRS. Ratios of nutrient range to reference laboratory method error were calculated and found to be positively related (R2 = 0.84) to NIRS predictability of a given TMR nutrient, suggesting some laboratory procedures were not precise enough to yield suitable NIRS predictions. Data suggest that NIRS has utility to predict basic nutrients such as CP, NDF, starch, NFC, and fat in TMR. However, difficulty was observed using NIRS in predicting key biological nutrients in TMR such as in situ CP fractions and in vitro NDF digestibility. Difficulty of NIRS in predicting these nutrients is related to the level of reference method error in relationship to the range of nutrient values in TMR, but other sources of prediction error may exist. PMID- 16702300 TI - Evaluation of the passage rate equations in the 2001 Dairy NRC model. AB - Dairy ration formulation to meet protein and amino acid requirements with the National Research Council Nutrient Requirements of Dairy Cattle (NRC, 2001) model depends on accuracy of predicting feed passage rates out of the rumen. The NRC (2001) passage rate (Kp) equations were evaluated for validity and sensitivity to input variables in predicting supplies of rumen degraded protein, rumen undegraded protein, and metabolizable protein. The database used in the development of the 3 Kp equations (for dry forage, wet forage, and concentrate) was used to independently derive the 3 equations using a meta-analysis technique. To extract quantitative relationships between statistically significant input variables and rate of passage, a random coefficients model that used each study effect as a random variable was used. The database was comprised of studies that only used rare earth markers. Outliers were identified by acceptance criteria defined a priori or the difference in fit statistic (DFFITS) value; 319, 63, and 139 treatment means were used to develop the Kp equations for dry forage, wet forage, and concentrate, respectively. We found that the sign of the regression coefficient for concentrate content in diet dry matter in the equation for Kp dry forage was inverted; it should be positive. A sensitivity analysis was conducted with a spreadsheet version of the NRC (2001) model developed for this study, using the Monte Carlo technique. The sensitivity analysis indicated that all Kp predictions were the most sensitive to variation in DM intake, and thus accurate measurement of DM intake is the most important factor in predicting Kp. Predictions for protein supply (rumen degraded protein, rumen undegraded protein, and metabolizable protein) were sensitive to variability in amount of feed crude protein (CP, %DM), digestion rate (Kd) of the B fraction of feed CP (%/h), and the Kp for concentrate (%/h), due to the high proportion of dietary CP in lactating dairy rations coming from concentrates. The sensitivity analysis indicated that accurate determinations of DMI, the Kd of the B fraction of feed CP, and feed CP are the most important variables needed to predict MP supply in lactating dairy cows with the NRC (2001) model. We conclude that the empirical Kp equations in the model are suitable for predicting passage rate in lactating dairy cows. More accurate predictions of Kp will require the development of a more mechanistic model that accounts for more of the biologically important variables (e.g., physical property of particles, liquid flow, and timely variation of intake) affecting passage rate. PMID- 16702301 TI - Distribution fitting and parameterization of individual operator work routine times for small dairy parlors. AB - A time and motion study was conducted at 13 small dairy farms with average herd sizes less than 100 cows. Parlors were configured with 3 to 6 stalls per side. A data acquisition methodology was developed using a video camera to gather work routine time data in the parlors. A computer-based data logger was used to extract individual event durations during video playback. Each parlor's video record was reviewed in the laboratory so that work routine times across all parlors and operators could be pooled to estimate typical operator performance. There were 34 operator work routine times associated with various procedures in milking parlors that were evaluated in this study. Individual times were compiled for each work routine and a data-fitting program called UNIFIT was used to fit the data to 1 of 4 data models: gamma, lognormal, Weibull, and Pearson #5. Each of 34 work routine variables was fitted, tested, and plotted to determine how well each of those models fit the actual data. Distributions for Pearson #5, lognormal, gamma, and Weibull models were best fitted to 12, 10, 8, and 4 work routine times, respectively. More common tasks such as attaching the milker, grabbing a towel, and drying the udder were more consistently executed and had smaller variances than routines in which the operator would leave the pit to go to the milk room or disassembled the milk collector after milking. One of the better fitting models was the lognormal distribution for the time to "attach milker," which had a low relative discrepancy to the P-P plot (model probability vs. data probability) of 0.019 and a moderate chi(2) test value of 0.358, thus demonstrating a good fit of the model to the data. Simulation tests were compared with observed data to validate models for work routine times and demonstrated that the models accurately predict parlor throughput in small- to medium-sized parlors. PMID- 16702303 TI - Antihypertensive peptides derived from egg proteins. AB - There have been studies of antihypertensive peptides derived from food proteins, but very few described the production of bioactive peptides from egg proteins. The first 2 antihypertensive peptides isolated in egg were obtained by enzymatic hydrolysis of ovalbumin. They correspond to the sequences Phe-Arg-Ala-Asp-His-Pro Phe-Leu (ovokinin) and Arg-Ala-Asp-His-Phe-Leu (ovokinin 2-7). Both exhibited endothelium-dependent vasodilatory activity. Ovokinin (2-7) had higher antihypertensive potency than ovokinin in spontaneously hypertensive rats (SHR). Modifications in the sequence of ovokinin (2-7) improved the bioavailability of this peptide. It was also demonstrated that different ovalbumin hydrolysates can inhibit angiotensin I-converting enzyme (ACE). We recently obtained an egg white hydrolysate that inhibited the enzyme in vitro. It was obtained by treating egg white with pepsin and it exhibited antihypertensive activity in SHR. Some ACE inhibitory peptides obtained from this hydrolysate (Tyr-Arg-Glu-Glu-Arg-Tyr-Pro Ile-Leu, Arg-Ala-Asp-His-Pro-Phe-Leu, and Ile-Val-Phe) also showed antihypertensive activity in these rats. The egg products mentioned could be used as functional food ingredients with potential therapeutic benefit in the prevention and treatment of hypertension. PMID- 16702302 TI - Bone, body weight, and weight reduction: what are the concerns? AB - Of the U.S. population, 65% is either overweight or obese, and weight loss is recommended to reduce co-morbid conditions. However, bone mobilization and loss may also occur with weight loss. The risk for bone loss depends on initial body weight, age, gender, physical activity, and conditions of dieting such as the extent of energy restriction and specific levels of nutrient intake. Older populations are more prone to bone loss with weight loss; in women, this is due at least in part to a reduced dietary Ca intake and/or efficiency of absorption. Potential hormonal mechanisms regulating bone loss during weight loss are discussed, including decreases in estrogen, leptin, glucagon-like peptide-2, growth hormone, and insulin-like growth factor-1, or an increase in cortisol. In contrast, the rise in adiponectin and ghrelin with weight reduction should not be detrimental to bone. Combining energy restriction with exercise does not necessarily prevent bone loss, but may attenuate loss as was shown with additional Ca intake or osteoporosis medications. Future controlled weight loss trials should be designed to further address mechanisms influencing the density and quality of bone sites vulnerable to fracture, in the prevention of osteoporosis. PMID- 16702304 TI - Glutamine pretreatment reduces IL-8 production in human intestinal epithelial cells by limiting IkappaBalpha ubiquitination. AB - Glutamine, the most abundant amino acid in the human body, plays several important roles in the intestine. Recent studies showed that glutamine regulates protein metabolism and intestinal inflammation among other mechanisms by reducing proinflammatory cytokine release. Because regulation of the inflammatory response was shown to be linked to proteolysis regulation, we hypothesized that glutamine pretreatment could act on IL-8 production in human intestinal epithelial cells through the regulation of inhibitor kappaB (IkappaB) ubiquitination. The HCT-8 cells were pretreated for 24 h with 0.6, 2, or 10 mmol/L glutamine. IL-8 concentration and IkappaB (free and ubiquitinated) expressions were assessed by ELISA and immunoblotting, respectively. A pretreatment with 10 mmol/L glutamine decreased IL-8 production under both basal and proinflammatory conditions (both P < 0.05). In the presence of a proteasome inhibitor (MG132), the ubiquitin IkappaBalpha complex expression was not significantly modified by glutamine under basal conditions but decreased significantly under proinflammatory conditions (P < 0.05). After the addition of 10 mmol/L of glutamine, the free IkappaBalpha expression increased under basal and stimulated conditions (both P < 0.05). A glutamine pretreatment of 10 mmol/L did not affect ubiquitin expression or proteasome activity. This study indicates that glutamine pretreatment may reduce the intestinal inflammatory response by limiting the proteolysis of IkappaBalpha. PMID- 16702305 TI - Regulation of protein synthesis by leucine starvation involves distinct mechanisms in mouse C2C12 myoblasts and myotubes. AB - Leucine modulates protein translation in higher eukaryotes by affecting phosphorylation and the function of proteins that regulate the initiation and/or elongation steps. These include the initiation factor 4E binding protein 1 (4E BP1), initiation factor 4E (eIF4E), initiation factor 2 (eIF2alpha), ribosomal S6 kinases (S6K1/2), and elongation factor 2 (eEF2). The alteration of protein translation by leucine starvation was studied during myogenic differentiation using the mouse C2C12 cell line as well as the role of rapamycin-sensitive mTOR (mammalian target of rapamycin) in the signaling of leucine in myotubes. A time course study showed that 1 h of leucine starvation decreased protein synthesis and S6K1 phosphorylation in myoblasts, whereas 3-5 h of starvation were necessary to induce such an alteration in myotubes. Although S6K1 phosphorylation was reduced in leucine-deprived myotubes, S6K2 and S6 phosphorylation were not affected. In contrast, rapamycin decreased the phosphorylation of S6K2 and S6 in myotubes. It is therefore likely that under the conditions present, the rapamycin sensitive mTOR was not affected by leucine starvation. S6K1 dephosphorylation may thus be mTOR independent, and the functional mTOR/S6K2 pathway may maintain S6 phosphorylation. An increased phosphorylation of eEF2 in myoblasts and myotubes indicated that global protein synthesis was reduced via a decrease in translation elongation. An increased association between 4E-BP1 and eIF4E, and increased phosphorylation of eIF2alpha also contributed to decreasing protein synthesis in leucine-starved myoblasts. In contrast, in leucine-starved myotubes, there were no change in the 4E-BP1-eIF4E association or eIF2alpha phosphorylation, suggesting that these factors were not rate limiting for decreasing protein synthesis in leucine-deprived myotubes. PMID- 16702306 TI - A rice bran oil diet increases LDL-receptor and HMG-CoA reductase mRNA expressions and insulin sensitivity in rats with streptozotocin/nicotinamide induced type 2 diabetes. AB - A rice bran oil (RBO) diet can reduce plasma lipids; this was attributed to the specific components, gamma-oryzanol and gamma-tocotrienol, which individually were shown to be hypocholesterolemic; however, the mechanism of their effects on diabetic hyperlipidemia and the development of diabetes is not known. Rats with streptozotocin/nicotinamide-induced type 2 diabetes were divided into control, RO10, and RO15 groups, and fed cholesterol-free diets containing 0, 10, and 15 g RBO with 0, 352, and 528 g gamma-oryzanol and 0, 6.0 and 9.0 mg gamma tocotrienol/100 g diet for 4 wk. Diabetic rats fed the RBO diet had greater insulin sensitivity (P = 0.02) than rats fed the control diet. Diabetic rats fed the RBO diet also had lower plasma triglyceride (P = 0.003), LDL cholesterol (P = 0.028), and hepatic triglyceride concentrations (P = 0.04), as well as greater fecal neutral sterol and bile acid excretion than those fed the control diet. After 4 wk, there was an approximately 100% (P < 0.001) increase in the abundance of hepatic cholesterol 7alpha-hydroxylase, an 89% (P < 0.001) increase in the hepatic LDL-receptor, and a 50% (P < 0.001) increase in hepatic 3-hydroxy-3 methylglutaryl coenzyme A reductase mRNA in rats fed the RBO diet compared with those fed the control diet. These findings support the conclusion that a rice bran oil-containing diet can significantly suppress hyperlipidemic and hyperinsulinemic responses in diabetic rats. The high contents of gamma-oryzanol and gamma-tocotrienol in RBO can lead to increased fecal neutral sterol and bile acid excretion, via upregulation of cholesterol synthesis and catabolism. PMID- 16702307 TI - Flavonoids and vitamin E reduce the release of the angiogenic peptide vascular endothelial growth factor from human tumor cells. AB - Neoangiogenesis is required for tumor development and progression. Many solid tumors induce vascular proliferation by production of angiogenic factors, prominently vascular endothelial growth factor (VEGF). Because nutrition is a causative factor for tumor prevention and promotion, we determined whether secondary plant constituents, i.e., flavonoids, tocopherols, curcumin, and other substances regulate VEGF in human tumor cells in vitro. VEGF release (concurrent with synthesis) from MDA human breast cancer cells and, for comparison, U-343 and U-118 glioma cells was measured by ELISA. Of 21 compounds tested, 9 showed significant inhibitory activity at 0.1 micromol/L in MDA human breast cancer cells. The rank order of inhibitory potency was naringin > rutin > alpha tocopheryl succinate (alpha-TOS) > lovastatin > apigenin > genistein > alpha tocopherol >or= kaempferol > gamma-tocopherol; chrysin and curcumin were inactive except at a concentration of 100 micromol/L. Glioma cells were similarly sensitive, with U343 more than U118, especially for alpha-TOS and tocopherols. Among the tocopherol derivatives, alpha-TOS (0.1 micromol/L) was the most effective in reducing VEGF release. Overall, the glycosylated flavonoids (i.e., naringin, a constituent of citrus fruits, and rutin, a constituent of cranberries) induced the greatest response to treatment at the lowest concentration in MDA human breast cancer cells. Inhibition of VEGF release by flavonoids, tocopherols, and lovastatin in these models of neoplastic cells suggests a novel mechanism for mammary cancer prevention. PMID- 16702308 TI - Bioproduction of conjugated linoleic acid by probiotic bacteria occurs in vitro and in vivo in mice. AB - Probiotics have been shown to reduce the incidence of colon cancer in animal models. The mechanisms responsible for this activity are poorly defined. Conjugated linoleic acids (CLA) are a group of isomers of linoleic acid (LA) possessing anti-inflammatory and anticarcinogenic properties, which can be produced from LA by certain bacterial strains. In this study, the ability of probiotic bacteria to exert anticarcinogenic effects through the production of CLA was assessed. Incubation of probiotic bacteria (VSL3, Lactobacillus acidophilus, L. bulgaricus, L. casei, L. plantarum, Bifidobacterium breve, B. infantis, B. longum, and Streptococcus thermophilus) in the presence of LA yielded CLA production as measured by gas chromatography. Conditioned medium, containing probiotic-produced CLA, reduced viability and induced apoptosis of HT 29 and Caco-2 cells, as assessed by MTT assay and DNA laddering, respectively. Western blotting demonstrated an increased expression of PPARgamma in cells treated with conditioned medium compared with LA alone. Incubation of murine feces with LA after administering VSL3 yielded 100-fold more CLA than feces collected prior to VSL3 feeding. This study supports a role for supplemental probiotics as a strategy both for attenuating inflammation and for preventing colon cancer. PMID- 16702309 TI - Plasma triglyceride and HDL-cholesterol concentrations in Vietnamese girls are affected by lipoprotein lipase, but not apolipoprotein CIII polymorphism. AB - Lipoprotein lipase (LPL) and apolipoprotein CIII (apoCIII) play an important role in HDL-cholesterol and triglyceride metabolism. This study examined the effects of LPL S447X and apoCIII SstI polymorphisms on the plasma lipid and lipoprotein concentrations in Vietnamese girls. Three hundred and fifty-one Vietnamese girls, from 7 to 9 y of age, were randomly selected from the urban and rural areas of Ho Chi Minh City, Vietnam. The presence of LPL S447X and apoCIII polymorphism was determined using the Invader assay, and the plasma lipid concentrations were measured by an enzymatic method. Dietary intakes and anthropometrics were assessed for each child. The frequencies of LPL 447X (11.9%) and apoCIII S2 (32.6%) polymorphisms in the Vietnamese were similar to those of other Asian populations. The frequency of LPL 447X was the same as that in Caucasians but the frequency of apoCIII S2 was approximately 3.8 times that in Caucasians. Overall, LPL 447X polymorphism was associated with a higher HDL-cholesterol concentration (7.4%, P = 0.007) and a lower triglyceride concentration (-13.6%, P = 0.04) than LPL 447S. The apoCIII S2 polymorphism was not associated with an increase in the plasma triglyceride concentration in Vietnamese girls with a low fat intake. However, due to the high frequency of the apoCIII SstI polymorphism and the increasingly westernized diet in Vietnam, attention should be paid to the interaction of genotype with the Vietnamese diet. PMID- 16702310 TI - Inhibition of betaine-homocysteine S-methyltransferase causes hyperhomocysteinemia in mice. AB - Inhibitors and methyl donor substrates for betaine-homocysteine S methyltransferase (BHMT) were used to study the role of this enzyme in the regulation of plasma total homocysteine (tHcy). Mice were administered an i.p. injection of S-(delta-carboxybutyl)-dl-homocysteine (CBHcy; 1 mg), a specific and potent inhibitor of BHMT, and tHcy and hepatic BHMT protein and activity levels were monitored over a 24-h period. Compared with saline-injected control mice, at 2 h postinjection, the CBHcy-treated mice had 87% lower BHMT activity and a 2.7 fold increase (11.1 vs. 3.0 micromol/L) in tHcy, effects that lasted nearly 8 h but returned to normal by 24 h. The level of BHMT protein remained constant over the 24-h period. After 6 CBHcy (1 mg) injections (one every 12 h), the mice had 7 fold higher tHcy, a 65% reduction in the liver S-adenosylmethionine:S adenosylhomocysteine ratio, and a marked upregulation of BHMT protein expression. At 2 h after injection of the sulfoxide derivative of CBHcy (10 mg) into mice, there was a modest reduction in BHMT activity and a 90% increase in tHcy. When given an injection of Met (3 mg) or Met plus CBHcy (1 mg), post-Met load tHcy levels were 2.2-fold higher (128 vs. 40 micromol/L) at 2 h postinjection in the mice given CBHcy. Like betaine, dimethylsulfoniopropionate was an effective tHcy lowering agent when given with a Met load. These studies are the first to show that transient inhibition of BHMT in vivo causes transient hyperhomocysteinemia, and that dimethylsulfoniopropionate can reduce a post-Met load rise in tHcy. PMID- 16702311 TI - The elevation of ingested lipids within plasma chylomicrons is prolonged in men compared with women. AB - The lipemic response to a high-fat meal is greater in men than in women. However, sex-related differences in the metabolic fate of ingested fat are not well understood. The purpose of this study was to measure the recovery of ingested fat in plasma fractions of chylomicrons (CHYLO), VLDL, and plasma fatty acids, as well as in expired breath (i.e., oxidation) in men and women. Nonobese subjects (n = 10; 5 men, 5 women) consumed 0.7 g fat/kg body weight containing 7 mg/kg of [1,1,1-13C]-trioleate the morning after an overnight fast. Plasma total triglyceride (TG) concentration and 13C recovery in the CHYLO, VLDL, and plasma fatty acid fractions, as well as expired breath samples, were measured over the 11-h period after the meal. Plasma total TG excursion was greater (P < 0.05) in men than in women during the 11-h period after the meal. Similarly, the recovery of the ingested tracer-labeled fat in the CHYLO fraction was greater in men than in women (main effect for sex; P < 0.05). Recovery of ingested tracer-labeled fat in VLDL, the plasma fatty acid fraction, and expired breath did not differ in men and women. Therefore, the elevated postprandial lipemia found in men compared with women was due to a prolonged availability of the lipid in chylomicrons, but was not related to differences in oxidation rates or incorporation of the ingested lipid into VLDL by the liver. PMID- 16702312 TI - The increase in human muscle protein synthesis induced by food intake is similar when assessed with the constant infusion and flooding techniques. AB - Food intake is accompanied by a stimulation of muscle protein synthesis. However, the reported magnitude of the response differs with different methods of measurement. The aim of this study was to assess whether the response to feeding is dependent on the technique used for measurement when length and amount of feeding are controlled. Muscle protein fractional synthesis rates (FSRs) were measured both in the fasting and feeding states in 2 groups of healthy volunteers (n = 8). Two techniques were used to measure FSR: in one group, FSRs were assessed with a primed constant infusion of L-[2H5]phenylalanine, whereas in the other, a flooding amount of the same label was employed. The fasting FSRs assessed with the constant infusion method and estimated using the free amino acid in the tissue fluid to represent the precursor pool for protein synthesis were comparable to those obtained with the flooding method (1.94 +/- 0.15 vs. 1.86 +/- 0.13%/d). The degree of stimulation due to feeding (P < 0.02) did not differ between the constant infusion (+15%) and flooding (+22%) techniques. The stimulatory effect of feeding on muscle FSR was associated with enhanced phosphorylation of the Mr = 70,000 ribosomal protein S6 kinase, suggesting that it may involve activation of translation. This study demonstrates that human muscle FSRs obtained with the constant infusion technique are comparable to those obtained with the flooding method and that, in response to feeding, the 2 techniques give comparable estimates of stimulation. PMID- 16702313 TI - The rate of intestinal glucose absorption is correlated with plasma glucose dependent insulinotropic polypeptide concentrations in healthy men. AB - Glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide (GIP) both play a role in the control of glucose homeostasis, and GIP is implicated in the regulation of energy storage. The capacity of carbohydrates to induce secretion of these incretin hormones could be one of the factors determining the metabolic quality of different types of carbohydrates. We analyzed the correlation between the rate of intestinal absorption of (starch derived) glucose and plasma concentrations of GLP-1 and GIP after ingestion of glucose and starchy foods with a different content of rapidly and slowly available glucose. In a crossover study, glucose, insulin, GLP-1, and GIP concentrations were monitored for 6 h after consumption of glucose, uncooked cornstarch (UCCS) or corn pasta in 7 healthy men. All test meals were naturally labeled with 13C. Using a primed, continuous D-[6,6-2H2]glucose infusion, the rate of appearance of exogenous glucose (RaEx) was estimated, reflecting the rate of intestinal glucose absorption. GLP-1 concentrations increased significantly from 180 to 300 min after ingestion of UCCS, the starch product with a high content of slowly available glucose. A high GIP response in the early postprandial phase (15-90 min) occurred after consumption of glucose. There was a strong positive within-subject correlation between RaEx and GIP concentrations (r = 0.73, P < 0.01) across the test meals. Rapidly and slowly digestible carbohydrates differ considerably in their ability to stimulate secretion of incretin hormones; the metabolic consequences of such differences warrant exploration. PMID- 16702314 TI - Luteolin and chrysin differentially inhibit cyclooxygenase-2 expression and scavenge reactive oxygen species but similarly inhibit prostaglandin-E2 formation in RAW 264.7 cells. AB - Inflammation and oxidative stress are associated with cancer, atherosclerosis, and other chronic diseases. Dietary flavonoids have been reported to possess antiinflammatory and antioxidant properties, but their mechanisms of action and structure-activity relations have not been fully investigated. We hypothesized that differences in antioxidant activity between the structurally similar flavones, luteolin and chrysin (differing only in B-ring hydroxylation patterns), would differentially affect inflammation-associated Cox-2 expression and PGE2 formation. Pretreatment of RAW 264.7 macrophage-like cells with 25, 50, or 100 micromol/L concentrations of luteolin inhibited lipopolysaccharide (LPS)-induced Cox-2 protein expression (P < 0.0001). Chrysin pretreatment did not reduce LPS induced Cox-2 protein expression at any level tested. Conversely, both luteolin and chrysin completely suppressed LPS-induced PGE2 formation (P < 0.001). Luteolin, but not chrysin, inhibited xanthine/xanthine oxidase-generated superoxide formation at 100 micromol/L in a cell-free system (P < 0.001). Although both luteolin and chrysin reduced LPS-induced hydroxyl radical formation relative to the positive control (P < 0.001), luteolin was superior to chrysin (P = 0.003). In summary, luteolin and chrysin suppressed PGE2 formation equally well, despite differential effects on Cox-2 protein expression and on superoxide and hydroxyl radical scavenging. These data indicate that flavones may display similar antiinflammatory activity via different mechanisms. PMID- 16702316 TI - The selenium metabolite methylselenol inhibits the migration and invasion potential of HT1080 tumor cells. AB - There is increasing evidence for the efficacy of certain forms of selenium as cancer-chemopreventive compounds. Methylselenol has been hypothesized to be a critical selenium metabolite for anticancer activity in vivo. To determine whether tumor cell migration, invasion, and cell cycle characteristics are inhibited by methylselenol, we exposed HT1080 cells to methylselenol. Methylselenol was generated with seleno-L-methionine (a substrate for methioninase). Submicromolar methylselenol exposure led to an increase in the G1 and G2 fractions with a concomitant drop in the S-phase, indicating slower cell growth. Furthermore, methylselenol inhibited the migration and invasion rate of the tumor cells by up to 53 and 76%, respectively, when compared with the control tumor cells. Although all cells had increased matrix metalloproteinase (MMP) enzyme activities of pro-MMP-2 and pro-MMP-9, the active form of MMP-2 was decreased in HT1080 cells cultured with methylselenol. In addition, methylselenol increased the protein levels of antimetastasic tissue inhibitor metalloproteinase (TIMP)-1 and TIMP-2. Collectively, these results demonstrate that submicromolar concentrations of methylselenol increase both prometastasis MMP-2 and MMP-9 and antimetastasis TIMP-1 and TIMP-2 expression. The apparent net effect of these changes is the inhibition of pro-MMP-2 activation and carcinogenic potential or activity. PMID- 16702315 TI - A folate- and methyl-deficient diet alters the expression of DNA methyltransferases and methyl CpG binding proteins involved in epigenetic gene silencing in livers of F344 rats. AB - Aberrations in methylation profile of the genome occur in human cancers induced by folate deficiency. To elucidate the underlying mechanism, male F344 rats were fed a diet deficient in l-methionine and devoid of folic acid and choline (FMD diet), which is known to induce hepatocellular carcinomas. We investigated changes in the DNA methylation machinery, namely, de novo DNA methyltransferases (Dnmt3a and 3b), maintenance DNA methyltransferase (Dnmt1), and methyl CpG binding proteins (MBDs), in rat livers during early stages of tumorigenesis. RT PCR and Western blot analyses revealed differential expression of these proteins in the livers of rats fed the FMD diet. Although the hepatic Dnmt1 mRNA level declined with age (P < 0.001), it was elevated (P < 0.001) in deficient rats compared with controls. The changes in hepatic Dnmt1 protein level with the diet correlated with its mRNA levels (r = 0.60, P = 0.002). Similarly, the Dnmt3a mRNA level was elevated in rats fed the FMD diet (P < 0.001), whereas the Dnmt3b level (mRNA and protein) was not affected by diet or age. Compared with controls, hepatic MBD1-3 RNA levels increased (P < 0.001) and the protein levels of MBD1, 2, and 4 were elevated (P < 0.001) in the deficient rats. In both diet groups, hepatic MBD2 protein decreased (P < 0.001), whereas MeCP2 protein increased (P < 0.001) with age. These results demonstrate that a combined folate and methyl deficiency alters components of the DNA methylation machinery by both transcriptional and posttranscriptional mechanisms during early stages of hepatocarcinogenesis. PMID- 16702317 TI - Carob pulp preparation rich in insoluble dietary fiber and polyphenols enhances lipid oxidation and lowers postprandial acylated ghrelin in humans. AB - Ghrelin is an orexigenic hormone that may affect substrate utilization in humans. Ghrelin is influenced by macronutrients, but the effects of insoluble dietary fiber and polyphenols are unknown. We investigated the effects of a polyphenol rich insoluble dietary fiber preparation from carob pulp (carob fiber) on postprandial ghrelin responses and substrate utilization. Dose-dependent effects of the consumption of carob fiber were investigated in a randomized, single blind, crossover study in 20 healthy subjects, aged 22-62 y. Plasma total and acylated ghrelin, triglycerides, and serum insulin and nonesterified fatty acids (NEFA) levels were repeatedly assessed before and after ingestion of an isocaloric standardized liquid meal with 0, 5, 10, or 20 g of carob fiber over a 300-min period. The respiratory quotient (RQ) was determined after consumption of 0 or 20 g of carob fiber. Carob fiber intake lowered acylated ghrelin to 49.1%, triglycerides to 97.2%, and NEFA to 67.2% compared with the control meal (P < 0.001). Total ghrelin and insulin concentrations were not affected by consumption of a carob fiber-enriched liquid meal. Postprandial energy expenditure was increased by 42.3% and RQ was reduced by 99.9% after a liquid meal with carob fiber compared with a control meal (P < 0.001). We showed that the consumption of a carob pulp preparation, an insoluble dietary fiber rich in polyphenols, decreases postprandial responses of acylated ghrelin, triglycerides, and NEFA and alters RQ, suggesting a change toward increased fatty acid oxidation. These results indicate that carob fiber might exert beneficial effects in energy intake and body weight. PMID- 16702318 TI - Maternal fish oil supplementation during lactation does not affect blood pressure, pulse wave velocity, or heart rate variability in 2.5-y-old children. AB - Maternal (n-3) PUFA deficiency is associated with higher blood pressure (BP) later in life in rat offspring, and early intake of (n-3) PUFA in formula-fed infants was shown to modify later BP. BP, heart rate (HR), and heart rate variability (HRV) are affected by dietary (n-3) PUFA in adults. In this study, we investigated whether fish oil (FO) supplementation of lactating mothers could modify BP, pulse wave velocity (PWV), and HRV in their children after 2 y. Mothers with low fish intake were randomly assigned to FO or olive oil (OO) supplementation for the first 4 mo after delivery. A reference group of mothers with a high habitual fish intake (HFI) was also followed. At the follow-up study at 2.5 y of age, BP and PWV were measured, and electrocardiograms were recorded for 0.5 h. FO supplementation significantly increased RBC levels of long-chain (n 3) PUFA of the 4 mo-old children, but at 2.5 y, the FO and OO groups did not differ. BP, PWV, HR, and HRV also did not differ among the groups. However, for all 3 groups, the children's intake of (n-3) PUFA at 2.5 y was negatively correlated with mean arterial pressure after adjustment for outdoor temperature (r = -0.245, P = 0.04). In conclusion, maternal FO supplementation had no overall effect on BP, PWV, or HRV of the children, indicating that (n-3) PUFA intake of Danish mothers may be sufficient in this sense. However, children's dietary intake of (n-3) PUFA might have a beneficial effect on BP in childhood. PMID- 16702319 TI - Dietary flaxseed supplementation ameliorates inflammation and oxidative tissue damage in experimental models of acute lung injury in mice. AB - Flaxseed (FS) is a nutritional supplement with high concentrations of (n-3) fatty acids and lignans that have anti-inflammatory and antioxidant properties. The use of FS in the prevention or treatment of acute lung disease is unknown. In this study, we evaluated diets with high FS content in experimental murine models of acute lung injury and inflammation. The kinetics of lignan accumulation in blood, following 10% FS supplementation, was determined using liquid chromatography tandem mass spectrometry. Mice were fed isocaloric control and 10% FS supplemented diets for at least 3 wk and challenged by hyperoxia (80% oxygen), intratracheal instillation of lipopolysaccharide, or acid aspiration. Bronchoalveolar lavage was evaluated for white blood cells, neutrophils, and proteins after a 24 h postintratracheal challenge of hydrochloric acid or lipopolysaccharide, or after 6 d of hyperoxia. Lung lipid peroxidation was assessed by tissue malondialdehyde concentrations. The plasma concentrations of the FS lignans, enterodiol and enterolactone, were stable after mice had eaten the diets for 2 wk. Following hyperoxia and acid aspiration, bronchoalveolar lavage neutrophils decreased in FS-supplemented mice (P = 0.012 and P = 0.027, respectively), whereas overall alveolar white blood cell influx tended to be lower (P = 0.11). In contrast, neither lung injury nor inflammation was ameliorated by FS following lipopolysaccharide instillation. Lung malondialdehyde levels were lower in hyperoxic mice than in unchallenged mice (P = 0.0001), and decreased with FS treatment following acid aspiration (P = 0.011). Dietary FS decreased lung inflammation and lipid peroxidation, suggesting a protective role against pro-oxidant-induced tissue damage in vivo. PMID- 16702320 TI - Apo-8'-lycopenal and apo-12'-lycopenal are metabolic products of lycopene in rat liver. AB - The health benefits of lycopene as an anticarcinogenic compound have been widely studied but little is known about the metabolic products of lycopene produced in vivo. We investigated lycopene metabolites in the liver of F344 male rats that had been prefed a lycopene-containing diet (0.25 g lycopene/kg diet). After 30 d of feeding, they were given a single oral dose of 14C-labeled lycopene (421.8 kBq). The metabolic products of both nonradioactive and 14C-labeled lycopene in rat liver were extracted and separated using HPLC and analyzed by UV/VIS spectrometry, online radioactive detection, and off-line and in-line positive ion electrospray ionization MS. Among a number of metabolite products formed, we identified apo-8'-lycopenal (lambdamax = 473 nm and m/z = 417). The putative compound, apo-12'-lycopenal, was detected but no apo-10'-lycopenal was present. A number of other very polar, short-chain and/or short chromophore compounds with UV/VIS absorption <300 nm were present but were not characterized. These data show that lycopene is cleaved in vivo by rats at different positions to produce apo-12'-lycopenal, and other unidentified metabolites in addition to apo-8' lycopenal. Apo-8'-lycopenal and the putative apo-12'-lycopenal are identified as lycopene metabolites in rat liver in vivo. PMID- 16702321 TI - Specific amino acids increase mucin synthesis and microbiota in dextran sulfate sodium-treated rats. AB - During the anabolic response associated with inflammation, mucin synthesis and colonic protection may be compromised by the limited availability of specific amino acids. We therefore determined the effect of dietary amino acid supplementation on the microbiota, mucin status, and mucosal damage in dextran sulfate sodium (DSS)-treated rats. From 8 d before to 28 d after colitis induction, male Sprague-Dawley rats (10 mo old, n = 8/group) were fed a control diet supplemented or not with 2 different doses of an amino acid cocktail containing L-threonine, L-serine, L-proline, and L-cysteine. All diets were isonitrogenous (adjusted with L-alanine). The higher dose of amino acids increased the number of Muc2-containing goblet cells in the surface epithelium of the ulcerated area, stimulated mucin production in the colon, and restored the mucin amino acid composition and mucosal content to healthy, control values. The colonic mucin synthesis rate was specifically stimulated by 95%, whereas the protein turnover was unchanged. All bacterial populations, markedly altered by the DSS treatment, were promoted. In conclusion, in inflammatory situations, an increase in threonine, serine, proline, and cysteine dietary supply can promote mucin synthesis, reequilibrate the gut microbiota, and thus favor colonic protection and mucosal healing. PMID- 16702323 TI - Maternal dietary (n-3) fatty acid deficiency alters neurogenesis in the embryonic rat brain. AB - Docosahexaenoic acid [22:6(n-3)] is enriched in brain membrane phospholipids and essential for brain function. Neurogenesis during embryonic and fetal development requires synthesis of large amounts of membrane phospholipid. We determined whether dietary (n-3) fatty acid deficiency during gestation alters neurogenesis in the embryonic rat brain. Female rats were fed diets with 1.3% energy [(n-3) control] or 0.02% energy [(n-3) deficient], from alpha-linolenic acid [18:3(n 3)], beginning 2 wk before gestation. Morphometric analyses were performed on embryonic day 19 to measure the mean thickness of the neuroepithelial proliferative zones corresponding to the cerebral cortex (ventricular and subventricular zones) and dentate gyrus (primary dentate neuroepithelium), and the thickness of the cortical plate and sectional area of the dentate gyrus. Phospholipids and fatty acids were determined by HPLC and GLC. Docosahexaenoic acid was 55-65% lower and (n-6) docosapentaenoic acid [22:5(n-6)] was 150-225% higher in brain phospholipids at embryonic day 19 in the (n-3) deficient (n = 6 litters) than in the control (n = 5 litters) group. The mean thickness of the cortical plate and mean sectional area of the primordial dentate gyrus were 26 and 48% lower, respectively, and the mean thicknesses of the cortical ventricular zone and the primary dentate neuroepithelium were 110 and 70% higher, respectively, in the (n-3) deficient than in the control embryonic day 19 embryos. These studies demonstrate that (n-3) fatty acid deficiency alters neurogenesis in the embryonic rat brain, which could be explained by delay or inhibition of normal development. PMID- 16702322 TI - Long-term ingestion of high flavanol cocoa provides photoprotection against UV induced erythema and improves skin condition in women. AB - Dietary antioxidants contribute to endogenous photoprotection and are important for the maintenance of skin health. In the present study, 2 groups of women consumed either a high flavanol (326 mg/d) or low flavanol (27 mg/d) cocoa powder dissolved in 100 mL water for 12 wk. Epicatechin (61 mg/d) and catechin (20 mg/d) were the major flavanol monomers in the high flavanol drink, whereas the low flavanol drink contained 6.6 mg epicatechin and 1.6 mg catechin as the daily dose. Photoprotection and indicators of skin condition were assayed before and during the intervention. Following exposure of selected skin areas to 1.25 x minimal erythemal dose (MED) of radiation from a solar simulator, UV-induced erythema was significantly decreased in the high flavanol group, by 15 and 25%, after 6 and 12 wk of treatment, respectively, whereas no change occurred in the low flavanol group. The ingestion of high flavanol cocoa led to increases in blood flow of cutaneous and subcutaneous tissues, and to increases in skin density and skin hydration. Skin thickness was elevated from 1.11 +/- 0.11 mm at wk 0 to 1.24 +/- 0.13 mm at wk 12; transepidermal water loss was diminished from 8.7 +/- 3.7 to 6.3 +/- 2.2 g/(h x m2) within the same time frame. Neither of these variables was affected in the low flavanol cocoa group. Evaluation of the skin surface showed a significant decrease of skin roughness and scaling in the high flavanol cocoa group compared with those at wk 12. Dietary flavanols from cocoa contribute to endogenous photoprotection, improve dermal blood circulation, and affect cosmetically relevant skin surface and hydration variables. PMID- 16702324 TI - A combined deficiency of vitamins E and C causes severe central nervous system damage in guinea pigs. AB - A short period of combined deficiency of vitamins E and C causes profound central nervous system (CNS) dysfunction in guinea pigs. For this report, CNS histopathology was studied to define the nature and extent of injury caused by this double deficiency. Weanling guinea pigs were fed a vitamin E-deficient or replete diet for 14 d. Then vitamin C was withdrawn from the diet of some guinea pigs. Four diet groups were thus formed: replete, vitamin E deficient, vitamin C deficient, and both vitamin E and C deficient. From 5 to 11 d after institution of the doubly deficient diet, 9 of 12 guinea pigs developed paralysis, and 2 more were found dead. The remaining guinea pig in the doubly deficient group and all animals in the other 3 groups survived without clinical impairment until the experiment was terminated at 13-15 d. Brains and spinal cords were serially sectioned and stained for examination. Only the combined deficiency produced damage in the CNS. The damage consisted mainly of nerve cell death, axonal degeneration, vascular injury, and associated glial cell responses. The spinal cord and the ventral pons in the brainstem were most severely affected, often exhibiting asymmetric cystic lesions. Several features of the lesions suggest that the primary damage was to blood vessels. These results indicate that the paralysis and death caused by combined deficiency of vitamins E and C in guinea pigs is caused by severe damage in the brainstem and spinal cord. PMID- 16702325 TI - Changes in the stability of dietary patterns in a study of middle-aged Swedish women. AB - Dietary patterns reflecting food habits may be associated with chronic diseases, yet little is known about the stability of these patterns. The objective of this study was to observe over time the stability of dietary patterns measured with exploratory and confirmatory factor analysis. Four random subsamples of 1000 women between 49 and 70 y old were chosen from >60,000 women included in the Swedish Mammography Cohort. Subjects in these subsamples were administered a FFQ 4, 5, 6, or 7 y after the baseline questionnaire; 3607 of the women responded (90% response rate). The stability of dietary patterns was evaluated with Spearman correlation coefficients between pattern scores at baseline and follow ups and by a test of internal stability, which evaluated the significance of changes within patterns between baseline and follow-up. We found 3 major dietary patterns: a healthy pattern, a Western pattern, and an alcohol pattern. Correlations between explored dietary pattern scores at baseline and at follow-up decreased from 0.59 (P < 0.01) after 4 y to 0.50 (P < 0.01) after 7 y for the healthy pattern, from 0.47 (P < 0.01) to 0.39 (P < 0.01) for the Western pattern and from 0.54 (P < 0.01) to 0.46 (P < 0.01) for the alcohol pattern. After 4 and 5 y, there was no evidence for internal instability in any of the 3 patterns. The Western pattern became internally unstable after 6 and 7 y and the alcohol pattern was unstable after 7 y. Our findings for this specific population suggest that in longitudinal studies, dietary exposures should be updated after at least 7 y. PMID- 16702326 TI - Selected diet and lifestyle factors are associated with estrogen metabolites in a multiracial/ethnic population of women. AB - Diet and lifestyle factors, body size, and smoking behavior may influence estrogen metabolism, but the nature of these relations may vary according to race/ethnic groups. We evaluated the association of lifestyle factors with estrogen metabolites 2-hydroxyestrone (2-OHE1) and 16alpha-hydroxyestrone (16alpha-OHE1) in a racially diverse population. With a cross-sectional study design, urine samples from 1881 African-American, Caucasian, Chinese, Japanese, and Hispanic women, aged 42-52 y, from the Study of Women's Health Across the Nation (SWAN) were assayed by EIA for 2-OHE1 and 16alpha-OHE1. Dietary factors and beverages were measured using a modified Block FFQ. Dietary fiber, vegetable and fruit servings, Brassica vegetables, polyphenols, coffee, caffeine, green and black tea, and total alcohol and wine were related to metabolite values using multiple variable regression analyses. In adjusted analyses, 2-OHE1 concentrations were significantly associated with race/ethnicity, weight, smoking, and consumption of hydroxybenzoic acid, anthocyanidins, wine, and caffeine (P < 0.05). Regression models incorporating these variables explained 19 20% of the variation in 2-OHE1 concentrations. Regression models for 16alpha OHE1, which explained 16-17% of the variability, included race/ethnicity, smoking, caffeine, total dietary fiber, and fiber from fruits and vegetables as variables. These associations may reflect why increased consumption of polyphenol containing foods and fruit as well as decreased smoking, caffeine intake, and body size would be consistent with hypothesized benefits and risks for selected health outcomes. PMID- 16702327 TI - CYP17 genotype modifies the association between lignan supply and premenopausal breast cancer risk in humans. AB - Cytochrome P450c17alpha (CYP17) has been associated with alterations in steroid hormone levels and premenopausal breast cancer risk and could modify the association between phytoestrogen intake and breast cancer risk. We examined plasma concentrations of enterolactone and genistein, estimated dietary phytoestrogen intake, CYP17 5'-UTR MspA1 genetic polymorphism, and breast cancer risk in 267 premenopausal breast cancer patients and 573 age-matched population controls from Germany. Multivariate logistic regression was used to estimate breast cancer risk associated with quartiles of phytoestrogen intake by genotype and to investigate gene-nutrient interactions. Premenopausal breast cancer risk was not significantly associated with the CYP17 A2 genotype. We observed a significant modifying effect of CYP17 genotype on plasma enterolactone-associated breast cancer risk (P for interaction < 0.01). Plasma enterolactone was significantly inversely related to breast cancer risk only in A2A2 carriers, showing odds ratios and 95% CI of 0.02 (0.00-0.41) and 0.01 (0.00-0.21) for the third and fourth quartiles vs. the lowest quartile, respectively. This inverse association was also found for the calculated enterolignan production as well as matairesinol intake. Compared with A1A1 carriers with the lowest enterolactone supply, the risk reduction associated with a high enterolactone supply resulted in a comparably decreased breast cancer risk for all genotypes. For genistein, no clear indication for a differential effect by CYP17 genotype was obtained. Our results suggest that CYP17 genotype modifies the protective effect of lignans on premenopausal breast cancer risk. Women homozygous for A2 allele benefit most from high plasma enterolactone concentrations and a high consumption of dietary precursors. PMID- 16702328 TI - Development of a glycemic index database for food frequency questionnaires used in epidemiologic studies. AB - Consumption of foods with a high glycemic index (GI) or glycemic load (GL) is hypothesized to contribute to insulin resistance, which is associated with increased risk of diabetes mellitus, obesity, cardiovascular disease, and some cancers. However, dietary assessment of GI and GL is difficult because values are not included in standard food composition databases. Our objective was to develop a database of GI and GL values that could be integrated into an existing dietary database used for the analysis of FFQ. Food GI values were obtained from published human experimental studies or imputed from foods with a similar carbohydrate and fiber content. We then applied the values to the Women's Health Initiative (WHI) FFQ database and tested the output in a random sample of previously completed WHI FFQs. Of the 122 FFQ line items (disaggregated into 350 foods), 83% had sufficient carbohydrate (>5 g/serving) for receipt of GI and GL values. The foods on the FFQ food list with the highest GL were fried breads, potatoes, pastries, pasta, and soft drinks. The fiber content of foods had very little influence on calculated GI or GL estimates. The augmentation of this FFQ database with GI and GL values will enable etiologic investigations of GI and GL with numerous disease outcomes in the WHI and other epidemiologic studies that utilize this FFQ. PMID- 16702329 TI - Lutein and eicosapentaenoic acid interact to modify iNOS mRNA levels through the PPARgamma/RXR pathway in chickens and HD11 cell lines. AB - Two experiments were conducted to investigate the effect of lutein and fat or eicosapentaenoic acid (EPA) interaction on inducible nitric oxide synthase (iNOS), PPARs alpha, beta, and gamma, and retinoic acid X receptor (RXR) alpha and gamma mRNA levels. In Expt. 1, macrophages were collected from broiler chicks fed 3 or 6% dietary fat (g/100 g) with 0, 25, and 50 mg lutein/kg feed for 23 d. In Expt. 2, using a 3 x 3 factorial, eicosapentaenoic acid (EPA) at 0, 15 and 50 micromol/L and lutein at 0, 10 and 100 micromol/L were applied to HD11 cell culture for 24 h. In both experiments, cells were stimulated with lipopolysaccharide before RNA isolation. Lutein interacted with fat in Expt. 1 and with EPA in Expt. 2 to affect mRNA levels of iNOS, PPARgamma, and RXRalpha in chicken macrophages and HD11 cells, respectively (P < 0.05). At 3% dietary fat or up to 15 micromol/L EPA in the medium, increasing lutein increased the iNOS mRNA. However, at 6% dietary fat or 50 micromol/L EPA, lutein did not cause a rise in iNOS mRNA. Increasing lutein in the medium from 0 to 100 micromol/L decreased iNOS mRNA. Increasing lutein with high fat (6%) or EPA (15 micromol/L EPA) increased PPARgamma and RXRalpha mRNA levels. Lutein increased PPARalpha mRNA levels in both macrophages (P < 0.01) and HD11 (P = 0.01) cells and RXRgamma (P < 0.01) mRNA levels in macrophages. GW9662, a PPARgamma antagonist, prevented (P < 0.01) the lutein-induced iNOS mRNA downregulation in HD11 cells. LG101208, a RXR antagonist, prevented (P < 0.01) iNOS upregulation induced by 10 micromol/L lutein and iNOS mRNA downregulation induced by 100 micromol/L lutein. We conclude that lutein and EPA interact through the PPARgamma and RXR pathways to modulate iNOS mRNA. PMID- 16702331 TI - HIV-1 infection alters the retinol-binding protein:transthyretin ratio even in the absence of the acute phase response. AB - The ratio of retinol-binding protein (RBP) to transthyretin (TTR) has been proposed as an indirect method with which to assess vitamin A status in the context of inflammation. Few studies have been conducted among adults, and none examined the effect of HIV-1 infection. Our goal was to assess the RBP:TTR ratio among adults, including the effects of HIV-1 and the acute phase response. We used data from a cross-sectional study of 600 Kenyan women, of whom 400 had HIV 1. The effect of vitamin A supplementation among the HIV-1-infected participants was subsequently assessed in a randomized trial. Among HIV-1-uninfected women without an acute phase response, a RBP:TTR cut-off value of 0.25 had approximately 80% sensitivity and specificity to detect vitamin A deficiency (retinol <0.70 micromol/L). No RBP:TTR cut-off value demonstrated both high sensitivity and specificity among HIV-1 infected women without evidence of inflammation. HIV-1 infection and advanced HIV-1 disease were associated with higher RBP:TTR ratios. The effect of HIV-1 was independent of the acute phase response, which also increased the RBP:TTR ratio. Serum retinol increased with vitamin A supplementation among those with a low RBP:TTR ratio, although the effect was small and was not present among those with concurrent inflammation. Thus, the RBP:TTR ratio has modest ability to predict vitamin A deficiency among healthy adults, but HIV-1 infection alters the ratio, even in the absence of the acute phase response. Our results raise questions about the utility of this measurement given the high prevalence of HIV-1 infection in areas where vitamin A deficiency is common. PMID- 16702330 TI - A multimicronutrient-fortified seasoning powder enhances the hemoglobin, zinc, and iodine status of primary school children in North East Thailand: a randomized controlled trial of efficacy. AB - Anemia and co-existing deficiencies of zinc, iron, iodine, and vitamin A occur among children in many developing countries including NE Thailand, probably contributing to impairments in growth, immune competence, and cognition. Sustainable strategies are urgently required to combat these deficiencies. We assessed the efficacy of a micronutrient-fortified seasoning powder served with a school lunch on reducing anemia and improving the micronutrient status of rural NE Thai children. Children (n = 569) aged 5.5-13.4y from 10 schools were randomly assigned to receive a seasoning powder either unfortified or fortified with zinc (5 mg), iron (5 mg), vitamin A (270 microg), and iodine (50 microg) (per serving) and incorporated into a school lunch prepared centrally and delivered 5 d/wk for 31 wk. Teachers monitored school lunch consumption. Baseline and final micronutrient status, hemoglobinopathies, and infection or inflammation were assessed from blood and urine samples. For the primary outcome, anemia (based on hemoglobin), no intervention effect was apparent (odds ratio: 1.02 95% CI: 0.69, 1.51) after adjustment for design strata. The odds of zinc (based on serum zinc) and urinary iodine deficiency in the fortified group were 0.63 (0.42, 0.94) and 0.52 (0.38, 0.71) times those in the unfortified group, respectively. Fortification had no effect on serum retinol (0.61: 0.25,1.51), ferritin (1.12: 0.43, 2.96), or mean red cell volume (1.16: 0.82, 1.64). Therefore, a micronutrient-fortified seasoning powder is a promising vehicle for improving zinc, iodine, and hemoglobin status, and its potential for incorporation into lunch programs in day care centers and schools in NE Thailand warrants investigation. PMID- 16702332 TI - Introduction to the 5th Amino Acid Assessment Workshop. AB - Amino acids (AAs) may be consumed at intakes above those that could be obtained from the normal diet, to promote health status in certain specific situations (e.g., sports training, aging). In this context, the relevant AAs may be used at high intake levels, which may in turn trigger adverse effects. There is little information on the adverse effects or pathophysiological consequences of excessive intakes of individual amino acids or mixtures. Hence, a series of workshops (named AAAW) are being organized to bring together experts in the fields of amino acid metabolism and nutritional effects, cell and molecular biology, toxicology, and regulatory issues and policy, with the aim of establishing a paradigm for the characterization of risks associated with specific intakes of amino acids by humans. The first 3 workshops covered general aspects concerning AAs (functions, risk characterization, differences in subpopulations, definition of surrogate markers, etc.). The fourth AAAW focused on branched-chain AAs. The articles in this supplement issue of the journal summarize the fifth workshop in the series, which focused on sulfur amino acids. PMID- 16702333 TI - The sulfur-containing amino acids: an overview. AB - Methionine, cysteine, homocysteine, and taurine are the 4 common sulfur containing amino acids, but only the first 2 are incorporated into proteins. Sulfur belongs to the same group in the periodic table as oxygen but is much less electronegative. This difference accounts for some of the distinctive properties of the sulfur-containing amino acids. Methionine is the initiating amino acid in the synthesis of virtually all eukaryotic proteins; N-formylmethionine serves the same function in prokaryotes. Within proteins, many of the methionine residues are buried in the hydrophobic core, but some, which are exposed, are susceptible to oxidative damage. Cysteine, by virtue of its ability to form disulfide bonds, plays a crucial role in protein structure and in protein-folding pathways. Methionine metabolism begins with its activation to S-adenosylmethionine. This is a cofactor of extraordinary versatility, playing roles in methyl group transfer, 5'-deoxyadenosyl group transfer, polyamine synthesis, ethylene synthesis in plants, and many others. In animals, the great bulk of S-adenosylmethionine is used in methylation reactions. S-Adenosylhomocysteine, which is a product of these methyltransferases, gives rise to homocysteine. Homocysteine may be remethylated to methionine or converted to cysteine by the transsulfuration pathway. Methionine may also be metabolized by a transamination pathway. This pathway, which is significant only at high methionine concentrations, produces a number of toxic endproducts. Cysteine may be converted to such important products as glutathione and taurine. Taurine is present in many tissues at higher concentrations than any of the other amino acids. It is an essential nutrient for cats. PMID- 16702334 TI - The nutritional relationship linking sulfur to nitrogen in living organisms. AB - Nitrogen (N) and sulfur (S) coexist in the biosphere as free elements or in the form of simple inorganic NO3- and SO4(2-) oxyanions, which must be reduced before undergoing anabolic processes leading to the production of methionine (Met) and other S-containing molecules. Both N and S pathways are tightly regulated in plant tissues so as to maintain S:N ratios ranging from 1:20 to 1:35. As a result, plant products do not adequately fulfill human tissue requirements, whose mean S:N ratios amount to 1:14.5. The evolutionary patterns of total body N (TBN) and of total body S (TBS) offer from birth to death sex- and age-related specificities well identified by the serial measurement of plasma transthyretin (TTR). Met is regarded as the most limiting of all indispensable amino acids (IAAs) because of its participation in a myriad of molecular, structural, and metabolic activities of survival importance. Met homeostasis is regulated by subtle competitive interactions between transsulfuration and remethylation pathways of homocysteine (Hcy) and by the actual level of TBN reserves working as a direct sensor of cystathionine-beta-synthase activity. Under steady-state conditions, the dietary intake of SO4(2-) is essentially equal to total sulfaturia. The recommended dietary allowances for both S-containing AAs allotted to replace the minimal obligatory losses resulting from endogenous catabolism is largely covered by Western customary diets. By contrast, strict vegans and low income populations living in plant-eating countries incur the risk of chronic N and Met dietary deficiencies causing undesirable hyperhomocysteinemia best explained by the downsizing of their TBN resources and documented by declining TTR plasma values. PMID- 16702335 TI - Mammalian cysteine metabolism: new insights into regulation of cysteine metabolism. AB - The mammalian liver tightly regulates its free cysteine pool, and intracellular cysteine in rat liver is maintained between 20 and 100 nmol/g even when sulfur amino acid intakes are deficient or excessive. By keeping cysteine levels within a narrow range and by regulating the synthesis of glutathione, which serves as a reservoir of cysteine, the liver addresses both the need to have adequate cysteine to support normal metabolism and the need to keep cysteine levels below the threshold of toxicity. Cysteine catabolism is tightly regulated via regulation of cysteine dioxygenase (CDO) levels in the liver, with the turnover of CDO protein being dramatically decreased when intracellular cysteine levels increase. This occurs in response to changes in the intracellular cysteine concentration via changes in the rate of CDO ubiquitination and degradation. Glutathione synthesis also increases when intracellular cysteine levels increase as a result of increased saturation of glutamate-cysteine ligase (GCL) with cysteine, and this contributes to removal of excess cysteine. When cysteine levels drop, GCL activity increases, and the increased capacity for glutathione synthesis facilitates conservation of cysteine in the form of glutathione (although the absolute rate of glutathione synthesis still decreases because of the lack of substrate). This increase in GCL activity is dependent on up regulation of expression of both the catalytic and modifier subunits of GCL, resulting in an increase in total catalytic subunit plus an increase in the catalytic efficiency of the enzyme. An important role of cysteine utilization for coenzyme A synthesis in maintaining cellular cysteine levels in some tissues, and a possible connection between the necessity of controlling cellular cysteine levels to regulate the rate of hydrogen sulfide production, have been suggested by recent literature and are areas that deserve further study. PMID- 16702336 TI - The effects of sulfur amino acid intake on immune function in humans. AB - No direct data exist on the influence of supranormal intakes of sulfur amino acids on immune function in humans. However 3 major products of sulfur amino acids, glutathione (GSH), homocysteine (Hcy), and taurine (Tau), influence, mainly, inflammatory aspects of the immune response in vitro and in vivo. Methionine intakes above approximately 1 g/d transiently raise plasma Tau, Hcy, and GSH. Tau and GSH ameliorate inflammation. Hcy has the opposite effect. A biphasic relation, between cellular GSH and CD4+ and CD8+ numbers occurs in healthy men. How changes in sulfur amino acid intake influence this phenomenon is unknown. In animals, high Tau intakes are antiinflammatory. How immune function in humans is affected is unknown. A positive relation between plasma neopterin (a marker of a Th-1-type immune response) and Hcy indicates that Hcy may play a part in inflammatory aspects of Parkinson's disease and aging. In vitro, Hcy, at concentrations seen following consumption of approximately 6 g L-methionine/d in adults, increases the interactions among T lymphocytes, monocytes, and endothelium. Whether a similar phenomenon occurs in vivo is unknown. Polymorphisms in the methylenetetrahydrofolate reductase gene are associated with raised plasma Hcy in young but not old subjects. The relation of this observation to immune function is unknown. The relationships among Hcy, inflammatory aspects of disease, and in vitro alterations in immune cell behavior create a cautionary note about supplementation of diets with l-methionine to raise intake above approximately 1 g/d. Studies directly linking methionine intake, genetics, plasma Hcy, Tau, and GSH and immune function are needed. PMID- 16702337 TI - Functions of sulfur-containing amino acids in lipid metabolism. AB - It is known that plasma lipid levels are controlled not only by dietary fat and carbohydrate but also by dietary protein and amino acids. Although it used to be thought that the source of protein was important, it is known that amino acid composition, amino acids themselves, and peptides from digested protein are more important than the protein source. Sulfur-containing amino acids (SAAs) are recognized to be some of the most potent modulators of lipid metabolism among amino acids. It has been demonstrated that SAAs have an increasing effect on HDL (high-density lipoprotein)-cholesterol and a decreasing effect on VLDL (very low density lipoprotein)-cholesterol. These data lead us to propose that SAAs have some beneficial functions against atherosclerotic diseases and metabolic syndrome. Relative availability of SAAs (RASAA) as well as the amount of SAAs in dietary protein would determine lipid metabolism. Therefore, we propose RASAA as a feasible index for improvement of lipid metabolism by amino acids. Although it is not clear how SAAs influence gene expression and lipid metabolism at a molecular level, SAAs change the metabolic pathway through transcriptional stimulation and posttranslational modification of regulatory proteins. PMID- 16702338 TI - Comparative species utilization and toxicity of sulfur amino acids. AB - Animal studies have shown that several methionine (Met) and cysteine (Cys) analogs or precursors have L-Met- and L-Cys-sparing activity. Relative oral bioavailability (RBV) values, with the L-isomer of Met and Cys set at 100% (isosulfurous basis), are near 100% for D-Met for animals but only about 30% for humans. Both the OH and keto analogs of Met have high RBV-sparing values, as does N-acetyl-L-Met (the D-isomer of acetylated Met has no bioactivity). L Homocysteine has an RBV value of about 65% for Met sparing in rats and chicks, but D-homocysteine has little if any Met-sparing activity. S-Methyl-L-Met can partially spare Met, but only when fed under dietary conditions of choline/betaine deficiency. Relative to L-Cys, high RBV values exist for L cystine, N-acetyl-L-Cys, L-homocysteine, L-Met, and glutathione, but D-cystine, the keto analog of Cys, L-cysteic acid, and taurine have no Cys-sparing activity. l-2-Oxothiazolidine-4-carboxylate has an RBV value of 75%, D-homocysteine 70%, and DL-lanthionine 35% as Cys precursors. Under dietary conditions of Cys deficiency and very low inorganic sulfate (SO4) ingestion, dietary SO4 supplementation has been shown to reduce the Cys requirement of several animal species as well as humans. Excessive ingestion of Met, Cys, or cystine has also been studied extensively in experimental animals, and these sulfur amino acids (SAA) are well established as being among the most toxic of all amino acids that have been studied. Even though Cys and its oxidized product (cystine) are equally efficacious at levels at or below their dietary requirements for maximal growth, Cys is far more toxic than cystine when administered orally in the pharmacologic dosing range. Isosulfurous (excess) levels of cystine, N-acetyl-L-Cys, or glutathione are far less growth depressing than L-Cys when 6 to 10 times the minimally required level of these SAA compounds are fed to chicks. PMID- 16702339 TI - Sparing of methionine requirements: evaluation of human data takes sulfur amino acids beyond protein. AB - The intimate relation between amino acids and protein and nitrogen requirements is well recognized. Nutrition research has focused on the capacity of food to meet the need for nitrogen and indispensable amino acids (IAA) and led to the conclusion that the quality, not just the quantity, of protein is critical. This is especially relevant in regard to the sulfur amino acids (SAA) methionine and cysteine because of the increased understanding of their relation to chronic diseases (e.g., cardiovascular disease, dementia, cirrhosis), immunomodulation, DNA transcription, and RNA translation. Considerable effort has been expended to determine whether and to what extent cysteine can spare the requirement for the IAA methionine. In vivo studies in humans generally concur that the dietary requirement of the SAA ranges between 13 and 16 mg.kg(-1).d(-1), but how much can be met by cysteine relative to methionine remains controversial. This review examines the current status of in vivo estimates of methionine and cysteine requirements in human adults and considers needs beyond what is necessary for protein synthesis. Factors influencing the utilization of methionine and cysteine, especially those conditions that lead to toxicity on the one hand or beneficial effects on the other, are discussed. Data on alternative dietary sources of methyl groups (e.g., betaine, choline, phosphatidylcholine, S adenosylmethionine, S-methylmethionine) or sulfur (e.g., N-acetylcysteine or L-2 oxothiazolidine-4-carboxylic acid) support a role for the SAA "beyond protein." Other pathways may influence the specific requirement for methionine and/or cysteine, especially when the person is challenged by disease, inadequate availability of food, or environmental stress. PMID- 16702340 TI - The in vivo sparing of methionine by cysteine in sulfur amino acid requirements in animal models and adult humans. AB - Sulfur amino acid metabolism has been receiving increased attention because of the link to chronic diseases such as cardiovascular disease, Alzheimer's disease, and diabetes. In addition, the role of cysteine and optimal intakes for physiological substrates such as glutathione are currently of considerable interest in human health. Although the dietary indispensability of methionine is not in question, the ability of cysteine to substitute for a portion of its requirement has been the topic of much debate. Methionine is often the most limiting amino acid in the diets of the developing world's population because of its low concentration in cereal grains. Therefore, the ability of cysteine to substitute for methionine requirement is not just biologically interesting; it is also of considerable economic and social importance. The primary goal of this review is to discuss the available evidence on the effect of cysteine substitution for methionine to meet the total sulfur amino acid requirement in adult humans, including an assessment of the methodological features of experiments with conflicting results. Assessment of the requirement experiments for amino acids with complex metabolism such as methionine and cysteine must begin with a careful definition of requirements and what substitution means. As a result of these definitions, a set of criteria for the intakes of methionine that will allow demonstration of the substitution effect have been developed. Some recent publications are assessed using these definitions and criteria, and a possible reason for the conflicting results in the literature is proposed. An approach to estimating tolerable upper intakes is also proposed. Research on in vivo sulfur amino acid metabolism in humans is tremendously difficult, and therefore, we do not wish to be overly critical of the high-quality work of the ambitious and highly intelligent men and women who have conducted various studies. Our goal is to objectively review the data for the reader in a logical and comprehensive manner and propose methods that may avoid difficulties in future studies. PMID- 16702341 TI - Adequate range for sulfur-containing amino acids and biomarkers for their excess: lessons from enteral and parenteral nutrition. AB - The adequacy range of dietary requirements of specific amino acids in disease states is difficult to determine. In health, several techniques are available allowing rather precise quantification of requirements based on growth of the organism, rises in plasma concentration, or increases in the oxidation of marker amino acids during incremental administration of the amino acid under study. Requirements may not be similar in disease with regard to protein synthesis or with regard to specific functions such as scavenging of reactive oxygen species by compounds including glutathione. Requirements for this purpose can be assessed only when such a function can be measured and related to clinical outcome. There is apparent consensus concerning normal sulfur amino acid (SAA) requirements. WHO recommendations amount to 13 mg/kg per 24 h in healthy adults. This amount is roughly doubled in artificial nutrition regimens. In disease or after trauma, requirements may be altered for methionine, cysteine, and taurine. Although in specific cases of congenital enzyme deficiency, prematurity, or diminished liver function, hypermethionemia or hyperhomocysteinemia may occur, SAA supplementation can be considered safe in amounts exceeding 2-3 times the minimal recommended daily intake. Apart from some very specific indications (e.g., acetaminophen poisoning), the usefulness of SAA supplementation is not yet established. There is a growing body of data pointing out the potential importance of oxidative stress and resulting changes in redox state in numerous diseases including sepsis, chronic inflammation, cancer, AIDS/HIV, and aging. These observations warrant continued attention for the potential role of SAA supplementation. In particular, N-acetylcysteine remains promising for these conditions. PMID- 16702342 TI - Sulfur amino acid metabolism in pregnancy: the impact of methionine in the maternal diet. AB - Animal studies show that the balance of methionine relative to other amino acids in the maternal diet is critical, as fetal growth is not only retarded by diets that are deficient but also by those containing excess. Diets with an inappropriate balance of methionine can adversely affect both short-term reproductive function and the long-term physiology of the offspring. The catabolism of unused methionine increases the demand for glycine and may cause a deficiency. High levels of methionine may also perturb intracellular S-adenosyl methionine pools and have an effect on the methylation of DNA and proteins. Excess methionine in the diet may also indirectly influence fetal development through the production of homocysteine or by the perturbation of endocrine functions. The metabolic interactions among dietary methionine, folic acid, and choline mean that other diet components can also change the methionine requirement. PMID- 16702343 TI - Assessing the effects of high methionine intake on DNA methylation. AB - Methylation of DNA occurs at cytosines within CpG (cytosine-guanine) dinucleotides and is 1 of several epigenetic mechanisms that serve to establish and maintain tissue-specific patterns of gene expression. The methyl groups transferred in mammalian DNA methylation reactions are ultimately derived from methionine. High dietary methionine intake might therefore be expected to increase DNA methylation. Because of the circular nature of the methionine cycle, however, methionine excess may actually impair DNA methylation by inhibiting remethylation of homocysteine. Although little is known regarding the effect of dietary methionine supplementation on mammalian DNA methylation, the available data suggest that methionine supplementation can induce hypermethylation of DNA in specific genomic regions. Because locus-specific DNA hypomethylation is implicated in the etiology of various cancers and developmental syndromes, clinical trials of "promethylation" dietary supplements are already under way. However, aberrant hypermethylation of DNA could be deleterious. It is therefore important to determine whether dietary supplementation with methionine can effectively support therapeutic maintenance of DNA methylation without causing excessive and potentially adverse locus-specific hypermethylation. In the viable yellow agouti (Avy) mouse, maternal diet affects the coat color distribution of offspring by perturbing the establishment of methylation at the Avy metastable epiallele. Hence, the Avy mouse can be employed as a sensitive epigenetic biosensor to assess the effects of dietary methionine supplementation on locus specific DNA methylation. Recent developments in epigenomic approaches that survey locus-specific DNA methylation on a genome-wide scale offer broader opportunities to assess the effects of high methionine intake on mammalian epigenomes. PMID- 16702344 TI - The 5th workshop on the assessment of adequate intake of dietary amino acids: general discussion 1. PMID- 16702345 TI - Screening of toxicity biomarkers for methionine excess in rats. AB - Although many animal studies have reported that dietary excess of methionine causes toxic changes including growth suppression and hemolytic anemia, the biochemical mechanism and biomarkers for methionine toxicity have not been well elucidated. The present study aimed to identify toxicity biomarkers from plasma metabolites in rats fed excessive methionine. Young growing rats were fed graded doses of additional methionine for 2 wk. Cluster analysis of multivariate correlations was performed on the physiological and toxicity variables with plasma metabolites detected by GC/MS, amino acid analyzer, and thiol-specific analysis. Indicative variables for hemolysis such as splenic nonheme iron content and plasma bilirubin were grouped in the same cluster as many methionine metabolites. Homocysteine and some undefined metabolites in this cluster were found to be strong discriminators between nontoxic and toxic levels of methionine intake. Product-to-precursor ratios of each methionine metabolite demonstrated that excessive methionine intake caused a marked decrease only in the ratio of cystathionine to homocysteine, suggesting that metabolism from homocysteine to cystathionine would be rate limiting in the disposal of excessive methionine. Collectively from these results, homocysteine appeared to be the most plausible biomarker to assess methionine excess as a surrogate marker both for toxicity and for setting a metabolic upper limit. PMID- 16702346 TI - Toxicity of methionine in humans. AB - The literature has been searched to identify evidence relating to the possible toxicity of the amino acid methionine in human subjects. Nutritional and metabolic studies have employed amounts of methionine, including the d and dl isomers, both below and above the requirement and have not reported adverse effects in adults and children. Although methionine is known to exacerbate psychopathological symptoms in schizophrenic patients, there is no evidence of similar effects in healthy subjects. The role of methionine as a precursor of homocysteine is the most notable cause for concern. A "loading dose" of methionine (0.1 g/kg) has been given, and the resultant acute increase in plasma homocysteine has been used as an index of the susceptibility to cardiovascular disease. Although this procedure results in vascular dysfunction, this is acute and unlikely to result in permanent damage. However, a 10-fold larger dose, given mistakenly, resulted in death. Longer-term studies in adults have indicated no adverse consequences of moderate fluctuations in dietary methionine intake, but intakes higher than 5 times normal resulted in elevated homocysteine levels. These effects of methionine on homocysteine and vascular function are moderated by supplements of vitamins B-6, B-12, C, and folic acid. In infants, methionine intakes of 2-5 times normal resulted in impaired growth and extremely high plasma methionine levels, but no adverse long-term consequences were observed. PMID- 16702347 TI - The many facets of hyperhomocysteinemia: studies from the Framingham cohorts. AB - Homocysteine is a sulfur amino acid whose metabolism stands at the intersection of 2 pathways: remethylation, which requires folic acid and B-12 coenzymes, and transsulfuration, which requires pyridoxal-5'-phosphate, the B-6 coenzyme. Data from several studies suggest that mild elevations of homocysteine in plasma are a risk factor for occlusive vascular disease. In the Framingham studies we have shown that plasma total homocysteine concentration is inversely related to the intake and plasma levels of folate and vitamin B-6 as well as vitamin B-12 plasma levels. Almost two-thirds of the prevalence of high homocysteine is attributable to low vitamin status or intake. Elevated homocysteine concentrations in plasma are a risk factor for prevalence of extracranial carotid artery stenosis of at least 25% in both men and women. Prospectively elevated plasma homocysteine is associated with increased total and CVD mortality, increased incidence of stroke, increased incidence of dementia and Alzheimer's disease, increased incidence of bone fracture, and higher prevalence of chronic heart failure. This multitude of relationships between elevated plasma total homocysteine and diseases that afflict the elderly point to the existence of a common denominator that may be responsible for these diseases. Whether this denominator is homocysteine itself or whether homocysteine is merely a marker remains to be determined. PMID- 16702348 TI - The Hordaland Homocysteine Study: a community-based study of homocysteine, its determinants, and associations with disease. AB - The Hordaland Homocysteine Study (HHS) is a population-based study of more than 18,000 men and women in the county of Hordaland in Western Norway. The first investigation (HHS-I) took place in 1992-93, when the subjects were aged 40-67 y. In 1997-99, a follow-up study (HHS-II) of 7,053 subjects was carried out. In this large population, plasma levels of total homocysteine (tHcy) are associated with several physiologic and lifestyle factors and common diseases. Increasing age, male sex, smoking, coffee consumption, high blood pressure, unfavorable lipid profile, high creatinine, and the MTHFR 677C > T polymorphism are among the factors associated with increased tHcy levels; physical activity, moderate alcohol consumption, and a good folate or vitamin B-12 status are associated with lower tHcy levels. Subjects with raised tHcy levels have increased risk of cardiovascular morbidity, cardiovascular and noncardiovascular mortality, and are more likely to suffer from depression and from cognitive deficit (elderly). Among women, raised tHcy levels are associated with decreased bone mineral density and increased risk of osteoporosis. Women with raised tHcy levels also have an increased risk of having suffered from pregnancy complications and an adverse pregnancy outcome. Significant associations between tHcy and clinical outcomes are usually observed for tHcy levels > 15 micromol/L, but for most conditions, there is a continuous concentration-response relation with no apparent threshold concentration. Overall, the findings from HHS indicate that a raised tHcy level is associated with multiple clinical conditions, whereas a low tHcy level is associated with better physical and mental health. PMID- 16702349 TI - Pathophysiological consequences of homocysteine excess. AB - Elevated level of the nonprotein amino acid homocysteine (Hcy) is a risk factor for cardiovascular diseases, neurodegenerative diseases, and neural tube defects. However, it is not clear why excess Hcy is harmful. To explain Hcy toxicity, the "Hcy-thiolactone hypothesis" has been proposed. According to this hypothesis, metabolic conversion of Hcy to a chemically reactive metabolite, Hcy-thiolactone, catalyzed by methionyl-tRNA synthetase is the first step in a pathway that contributes to Hcy toxicity in humans. Plasma Hcy-thiolactone levels are elevated in human subjects with hyperhomocysteinemia caused by mutations in CBS or MTHFR genes. Plasma and urinary Hcy-thiolactone levels are also elevated in mice fed a high-methionine diet. Hcy-thiolactone can be detrimental because of its intrinsic ability to form N-Hcy-protein adducts, in which a carboxyl group of Hcy is N linked to epsilon-amino group of a protein lysine residue. This article reviews recent studies of Hcy-thiolactone and N-Hcy-protein in the human body, including their roles in autoimmune response, cellular toxicity, and atherosclerosis. Potential utility of Hcy-thiolactone, N-Hcy-protein, or anti-N-Hcy-protein autoantibodies as markers of Hcy excess is discussed. PMID- 16702350 TI - Inborn errors of sulfur-containing amino acid metabolism. AB - Two superimposed metabolic sequences, transsulfuration and the methionine/homocysteine cycle, form the pathway for methionine metabolism in mammalian liver. This combined pathway was formulated first to explain observations in subjects with homocystinuria caused by cystathionine synthase deficiency. Since that time additional inborn errors have been discovered, and currently we know of human subjects with isolated defects in all of the reactions of the combined pathway with only one exception: betaine homocysteine methyltransferase. Studies of these inborn errors have contributed significantly to our knowledge of human methionine metabolism and to the clinical consequences of impaired metabolism. Transsulfuration appears to function primarily for the metabolism of excess methionine, and each of the 5 defects in this pathway results in the accumulation of 1 or more of the normal metabolites. Thus, studies of these disorders may provide insight into both the potential pathological sequelae of nutritional methionine excess as well as whether laboratory testing allows the detection of excess. PMID- 16702351 TI - The 5th workshop on the assessment of adequate intake of dietary amino acids: general discussion 2. PMID- 16702352 TI - Isochrony in the olivocerebellar system underlies complex spike synchrony. PMID- 16702353 TI - The antibacterial activity of human neutrophils and eosinophils requires proton channels but not BK channels. AB - Electrophysiological events are of central importance during the phagocyte respiratory burst, because NADPH oxidase is electrogenic and voltage sensitive. We investigated the recent suggestion that large-conductance, calcium-activated K(+) (BK) channels, rather than proton channels, play an essential role in innate immunity (Ahluwalia, J., A. Tinker, L.H. Clapp, M.R. Duchen, A.Y. Abramov, S. Page, M. Nobles, and A.W. Segal. 2004. Nature. 427:853-858). In PMA-stimulated human neutrophils or eosinophils, we did not detect BK currents, and neither of the BK channel inhibitors iberiotoxin or paxilline nor DPI inhibited any component of outward current. BK inhibitors did not inhibit the killing of bacteria, nor did they affect NADPH oxidase-dependent degradation of bacterial phospholipids by extracellular gIIA-PLA(2) or the production of superoxide anion (O(2*)(-)). Moreover, an antibody against the BK channel did not detect immunoreactive protein in human neutrophils. A required role for voltage-gated proton channels is demonstrated by Zn(2+) inhibition of NADPH oxidase activity assessed by H(2)O(2) production, thus validating previous studies showing that Zn(2+) inhibited O(2*)(-) production when assessed by cytochrome c reduction. In conclusion, BK channels were not detected in human neutrophils or eosinophils, and BK inhibitors did not impair antimicrobial activity. In contrast, we present additional evidence that voltage-gated proton channels serve the essential role of charge compensation during the respiratory burst. PMID- 16702354 TI - Mechanisms underlying heterogeneous Ca2+ sparklet activity in arterial smooth muscle. AB - In arterial smooth muscle, single or small clusters of Ca(2+) channels operate in a high probability mode, creating sites of nearly continual Ca(2+) influx (called "persistent Ca(2+) sparklet" sites). Persistent Ca(2+) sparklet activity varies regionally within any given cell. At present, the molecular identity of the Ca(2+) channels underlying Ca(2+) sparklets and the mechanisms that give rise to their spatial heterogeneity remain unclear. Here, we used total internal reflection fluorescence (TIRF) microscopy to directly investigate these issues. We found that tsA-201 cells expressing L-type Cavalpha1.2 channels recapitulated the general features of Ca(2+) sparklets in cerebral arterial myocytes, including amplitude of quantal event, voltage dependencies, gating modalities, and pharmacology. Furthermore, PKCalpha activity was required for basal persistent Ca(2+) sparklet activity in arterial myocytes and tsA-201 cells. In arterial myocytes, inhibition of protein phosphatase 2A (PP2A) and 2B (PP2B; calcineurin) increased Ca(2+) influx by evoking new persistent Ca(2+) sparklet sites and by increasing the activity of previously active sites. The actions of PP2A and PP2B inhibition on Ca(2+) sparklets required PKC activity, indicating that these phosphatases opposed PKC-mediated phosphorylation. Together, these data unequivocally demonstrate that persistent Ca(2+) sparklet activity is a fundamental property of L-type Ca(2+) channels when associated with PKC. Our findings support a novel model in which the gating modality of L-type Ca(2+) channels vary regionally within a cell depending on the relative activities of nearby PKCalpha, PP2A, and PP2B. PMID- 16702356 TI - Plasmalemmal Ca2+ signaling in arterial smooth muscle: it's elementary! PMID- 16702355 TI - Ca2+-activated Cl- current from human bestrophin-4 in excised membrane patches. AB - Bestrophins are a newly discovered family of Cl(-) channels, some members of which are activated by intracellular Ca(2+). So far, all studies were carried out with whole-cell recordings from plasmid-transfected cultured cells, so it is unclear whether Ca(2+) activates bestrophin through a metabolic mechanism or in a more direct way. We report here experiments that addressed this question with excised, inside-out membrane patches. We chose human bestrophin-4 (hBest4) for heterologous expression because it gave particularly large Cl(-) currents when expressed, thus allowing detection even in excised membrane patches. hBest4 gave a negligible Cl(-) current in a Ca(2+)-free solution on the cytoplasmic (bath) side, but produced a Cl(-) current that was activated by Ca(2+) in a dose dependent manner, with a K(1/2) of 230 nM. Thus, Ca(2+) appears to activate the bestrophin Cl(-) channel without going through a freely diffusible messenger or through protein phosphorylation. Because the activation and deactivation kinetics were very slow, however, we cannot exclude the involvement of a membrane associated messenger. PMID- 16702357 TI - Mathematical modeling: epidemiology meets systems biology. PMID- 16702358 TI - Understanding population and individual risk assessment: the case of polychlorinated biphenyls. AB - Decisions about how to improve or protect the public health can be, and sometimes necessarily are, made on imprecise science. The regulation of potential human carcinogens in the environment entails a population-risk assessment process intended to reduce risks to less than one additional cancer in 100,000 or 1,000,000 persons. These risk assessment processes, however, may be miscommunicated or misinterpreted in the context of individual cancer risks by scientists, regulators, the lay media, and the public. This commentary will review methods for establishing a causal relationship between carcinogen exposures and cancer risk. It will use the case of polychlorinated biphenyls (PCB) as an example of how to place scientific data into the context of human exposure and cancer risk. PCBs are widespread environmental contaminants and most people have detectable levels of PCBs in their bodies. The primary source for exposure in the general population is through the diet. PCBs are carcinogens in experimental animal models, but how this information can be extrapolated to human risk remains uncertain. PCB experimental studies provide data that are used to regulate and control human exposure, although the epidemiologic evidence fails to establish PCBs as human carcinogens. Thus, what is used for population-risk assessment may not be appropriate for individual-risk assessment or concluding that a causal relationship exists between PCB exposure and cancer risk. The hazards from a carcinogen designated by regulatory and review agencies as a "probable" human carcinogen is often misunderstood out of context about the magnitude of the risk and in what settings. How scientists communicate their results in scientific articles can strongly influence how others interpret their data. Misunderstandings from both the use of regulatory and review-agency opinions and the conclusions espoused by scientists occur in the media, among private physicians counseling their patients about cancer risk, and in the legal settings where plaintiffs seek compensation for exposure and alleged harm (or future harm). This can lead to false conclusions about what caused a cancer in a specific patient, undue anxiety about future cancer risk, inappropriate cancer screening, and attendant increased morbidity due to increased uses of the medical system and complication rates from medical procedures. The communication of research findings by scientists must be presented with caution, resisting the temptation to extrapolate, inappropriately, research data to the general population. PMID- 16702359 TI - Uptake rates for breast cancer genetic testing: a systematic review. AB - PURPOSE: Individuals and families dealing with the possibility of hereditary cancer risk face numerous decisions, including whether to obtain genetic testing. The purpose of this article is to determine what is known about the rate at which people obtain cancer genetic testing. METHODS: Using MEDLINE, CINAHL, and PSYCHINFO plus reviewing reference lists of relevant articles, we identified 40 studies in May 2002 that addressed breast cancer-related decisions, enrolled adult participants, were published in 1990 or more recently, were peer-reviewed primary clinical studies, addressed genetic testing either alone or in combination with genetic counseling, and reported rates at which participants showed interest in and/or underwent cancer genetic testing. Information regarding study design, participants, and genetic testing uptake rates was recorded. Each article was reviewed for methodologic quality using a flexible quality review system applicable to all study types. RESULTS: Of the 40 studies, 25 provided information about hypothetical genetic testing decisions, 14 about real decisions, and 1 about both. Mean hypothetical uptake was 66% (range, 20-96%) and real uptake was 59% (range, 25-96%). Multivariate logistic regression analyses found that decision type (real/hypothetical), personal and family history of breast cancer, and variability in sampling strategy, recruitment setting, and criteria for real and hypothetical uptake were independently associated with uptake. Our systematic review identified additional explanations for uptake variability (investigator influences, small sample sizes, variability in target populations, lack of clearly described sampling strategies, sampling methods open to bias, and variability in reporting associated risk factors). CONCLUSION: In addition to clinical characteristics, research methodologic issues are likely to be major determinants of variability in published breast cancer genetic testing uptake rates. An understanding of these issues will clarify to clinicians why their clinical experience may not be congruent with published rates and help guide future research. PMID- 16702360 TI - Difference in the incidence trend of nasopharyngeal and oropharyngeal carcinomas in Taiwan: implication from age-period-cohort analysis. AB - Lifestyle factors are considered important for the pathogenesis of both nasopharyngeal and oropharyngeal carcinomas. In Taiwan, the incidence of nasopharyngeal carcinoma gradually decreased over the past 20 years, whereas that of oropharyngeal carcinoma increased rapidly. To compare the incidence trends of nasopharyngeal and oropharyngeal carcinomas in Taiwan, the age-period-cohort model was used to analyze epidemiologic data from 1981 to 2000 obtained from the Taiwan Cancer Registry. The calendar time period of 1986 to 1990 and the 1931 to 1940 birth cohort were used as reference groups for estimates of relative risk. For nasopharyngeal carcinoma, the incidence seemed to decrease in most age groups and was more prominent in women (30%) than in men (23%). For oropharyngeal carcinoma, the incidence increased in all age groups and was more prominent in men (391.4%) than in women (59.2%). Cohort effect was found for both nasopharyngeal and oropharyngeal carcinomas. The relative risk of nasopharyngeal carcinoma for the 1971 to 1980 birth cohort was 0.38 for women and 0.68 for men. The relative risk of oropharyngeal carcinoma for the 1971 to 1980 cohort was 45.67 for men and 2.69 for women. Change in lifestyle seemed to be an important factor for the difference in the incidence trend between nasopharyngeal and oropharyngeal carcinomas and between men and women. PMID- 16702361 TI - Sequence variants in toll-like receptor 10 are associated with nasopharyngeal carcinoma risk. AB - Nasopharyngeal carcinoma (NPC) is a common malignancy in southern China and Southeast Asia. Genetic susceptibility is a major factor in determining the individual risk of NPC in these areas. To test the association between NPC and variants in Toll-like receptor 10 (TLR10), we conducted a hospital-based case control study in a Cantonese-speaking population in Guangdong province. Seven single nucleotide polymorphisms in TLR10, selected with a tagging algorithm, were genotyped. When assessing each unique haplotype compared with the most common haplotype, "GAGTGAA," with the expectation-maximization algorithm in Haplo.stats, the risk of developing NPC was significantly elevated among men who carried the haplotype "GCGTGGC" (P = 0.005). After adjusting for age, gender, and VCA-IgA antibody titers, this association was more significant (P = 0.0007). To further assess the overall differences of haplotype frequency profiles between cases and healthy controls, the global score test, considering all haplotypes and adjusting for age, gender, and VCA-IgA antibody titers, gave a haplo score of 27.52 with P = 0.002. The haplotype specific odds ratio was 2.66 (confidence interval, 1.34 3.82) for GCGTGGC. We concluded that in this Cantonese population-based study, haplotype GCGTGGC with frequency of 11.4% in TLR10 was found to be associated with NPC and this association was statistically significant after adjusting for age, gender, and VCA-IgA antibody titers. It is possible that this is not a causal haplotype for NPC; rather, it is in strong linkage disequilibrium with a causal single nucleotide polymorphism in close proximity. PMID- 16702362 TI - Perinatal risk factors for cancer of the esophagus and gastric cardia: a nested case-control study. AB - BACKGROUND: We have previously hypothesized that preterm birth or impaired fetal growth may cause esophageal adenocarcinomas through gastroesophageal reflux early in life. In this study, we aimed to test if there is an association between gestational duration and birth weight on the one hand, and risk of esophageal and cardia adenocarcinoma on the other. METHODS: We conducted a nested case-control study of 67 cases of esophageal adenocarcinoma and 93 cases of cardia adenocarcinoma, whereas 50 cases of squamous cell carcinoma were studied for comparison. Birth records of cases were traced. Three matched controls per case were randomly selected. Perinatal data were extracted from birth records. RESULTS: Long gestational duration was associated with a decreased risk of cardia adenocarcinoma (P(trend) = 0.001) and a nonsignificant decreased risk of esophageal adenocarcinoma (P = 0.07), whereas no such association was found for esophageal squamous cell carcinoma (P = 0.96). Birth weight was not associated with risk of any of the studied cancers. Compared with lower maternal age (25) was associated with an increased risk of esophageal adenocarcinoma (males, OR, 2.2; 95% CI, 1.7-2.7; females, OR, 2.0; 95% CI, 1.4-2.9). Higher levels of BMI were associated with increased risk (overweight males, OR, 1.8; 95% CI, 1.5-2.2; obese males, OR, 2.4; 95% CI, 1.9-3.2). The overall associations with cardia cancer were heterogeneous, although stratification by study location provided homogeneous results for populations from the United States or Europe. A high BMI was weakly associated with the risk of cardia adenocarcinoma (OR, 1.5; 95% CI, 1.3-1.8; P(heterogeneity) = 0.38). CONCLUSIONS: Pooled results from observational studies support a positive association between high BMI and the risk for esophageal and possibly for cardia adenocarcinoma. PMID- 16702364 TI - Anthropometry, physical activity, and the risk of pancreatic cancer in the European prospective investigation into cancer and nutrition. AB - Tobacco smoking is the only established risk factor for pancreatic cancer. Results from several epidemiologic studies have suggested that increased body mass index and/or lack of physical activity may be associated with an increased risk of this disease. We examined the relationship between anthropometry and physical activity recorded at baseline and the risk of pancreatic cancer in the European Prospective Investigation into Cancer and Nutrition (n = 438,405 males and females age 19-84 years and followed for a total of 2,826,070 person-years). Relative risks (RR) were calculated using Cox proportional hazards models stratified by age, sex, and country and adjusted for smoking and self-reported diabetes and, where appropriate, height. In total, there were 324 incident cases of pancreatic cancer diagnosed in the cohort over an average of 6 years of follow up. There was evidence that the RR of pancreatic cancer was associated with increased height [RR, 1.74; 95% confidence interval (95% CI), 1.20-2.52] for highest quartile compared with lowest quartile (P(trend) = 0.001). However, this trend was primarily due to a low risk in the lowest quartile, as when this group was excluded, the trend was no longer statistically significant (P = 0.27). A larger waist-to-hip ratio and waist circumference were both associated with an increased risk of developing the disease (RR per 0.1, 1.24; 95% CI, 1.04-1.48; P(trend) = 0.02 and RR per 10 cm, 1.13; 95% CI, 1.01-1.26; P(trend) = 0.03, respectively). There was a nonsignificant increased risk of pancreatic cancer with increasing body mass index (RR, 1.09; 95% CI, 0.95-1.24 per 5 kg/m(2)), and a nonsignificant decreased risk with total physical activity (RR, 0.82; 95% CI, 0.50-1.35 for most active versus inactive). Future studies should consider including measurements of waist and hip circumference, to further investigate the relationship between central adiposity and the risk of pancreatic cancer. PMID- 16702366 TI - Incidence of colorectal cancer in relation to glycemic index and load in a cohort of women. AB - BACKGROUND: Dietary glycemic index (GI) and glycemic load (GL) affect circulating insulin concentrations. Elevated circulating insulin concentrations can increase insulin-like growth factor-1, and both of these hormones may have growth promoting effects within the colorectum. METHODS: We examined associations of GI and GL with colorectal cancer (CRC) among participants in the Iowa Women's Health Study (n = 35,197; ages 55-69 years at baseline in 1986). Over 15 years of follow up, we identified 757 cases of colon cancer and 209 cases of rectal cancer (954 CRC cases). RESULTS: Overall, neither GI nor GL were significantly associated with incident CRC. However, among obese women (baseline body mass index >/=30 kg/m(2)) CRC incidence was increased in the highest versus lowest quintiles of GI (relative risk, 1.66; 95% confidence intervals, 1.13-2.43; P for trend = 0.02) and GL (relative risk, 1.79; 95% confidence intervals, 1.19-2.70; P for trend < 0.01). This pattern of increased risk for obese women with high GI or GL tended to hold for both colon cancer and rectal cancer, and for nondiabetic women as well. No statistically significant associations were observed between GI or GL and CRC among subjects whose baseline body mass index was <30 kg/m(2). CONCLUSION: Our findings suggest that high GI or GL are not major CRC risk factors among older women in general, but may increase CRC risk among women who are obese. PMID- 16702365 TI - DNMT3b polymorphism and hereditary nonpolyposis colorectal cancer age of onset. AB - Hereditary nonpolyposis colorectal cancer (HNPCC) is an autosomal dominant syndrome of familial malignancies resulting from germ-line mutations in DNA mismatch repair genes. Colorectal and endometrial cancers are most frequently observed. A polymorphic C-to-T change in the promoter region of the DNMT3b gene, 149 bp from the transcription start site, is reported to greatly increase promoter activity and is associated with increased risk for lung cancer and decreased postsurgical survival in patients with small cell carcinoma of the head and neck. We studied the influence of this DNMT3b polymorphism on HNPCC age of onset. We determined the DNMT3b genotype of 146 mismatch repair mutation carriers from 72 families. Of these, 74 participants had colorectal cancer. The participants were genotyped by single-strand conformational polymorphism analysis and DNA sequencing. We tested the association between age of onset and DNMT3b genotypes by comparing Kaplan-Meier survival curves, evaluating the homogeneity of the curves using the log-rank test, Wilcoxon's test, and Fleming-Harrington test and estimating the strength and direction of the association using the Cox proportional hazards regression model adjusting for potential demographic and genetic confounding factors. HNPCC patients carrying one or two copies of the DNMT3b variant T allele developed their colorectal cancer significantly earlier than HNPCC patients who were homozygous for the wild-type DNMT3b allele. Combining knowledge of an individual's DNMT3b genotype with information on other genetic and environmental risk factors may improve risk estimates and help to identify individuals who are genetically susceptible to developing HNPCC at an earlier age. PMID- 16702367 TI - Tobacco smoking and colorectal hyperplastic and adenomatous polyps. AB - Colorectal adenomas and possibly some hyperplastic polyps are precursors of colorectal cancer. Tobacco use is associated in epidemiologic studies with these polyps, although links between smoking and colorectal cancer are less consistent. To characterize the role of tobacco in early colorectal carcinogenesis, we compared tobacco use among 4,383 subjects with histologically verified benign (hyperplastic or adenomatous) polyps of the distal colon (descending colon, sigmoid, and rectum) with tobacco use among 33,667 subjects who were endoscopy negative for distal colon tumors, in the screening arm of the Prostate, Lung, Colorectal, and Ovarian Trial, a randomized trial of flexible sigmoidoscopy. Risks, estimated by the odds ratio (OR), associated with current cigarette use were OR = 4.4 [95% confidence interval (95% CI), 3.7-5.2] for hyperplastic polyps only, OR = 1.8 (95% CI, 1.5-2.1) for adenomas only, and OR = 6.2 (95% CI, 4.7 8.3) for subjects with both hyperplastic and adenomatous polyps concurrently. Effects were weaker among ex smokers; the smoking-associated ORs remained consistently higher for hyperplastic polyps. This pattern was also seen in relation to cigarettes smoked per day, smoking duration, and pack-years. Tobacco associated risks for multiple polyps were also stronger when hyperplastic disease was involved. In conclusion, tobacco use, particularly recent use, increases risk for both adenomatous and hyperplastic polyps, but the risks are substantially greater for hyperplastic lesions. PMID- 16702368 TI - Gender-specific protective effect of hemoglobin on arsenic-induced skin lesions. AB - Chronic arsenic poisoning remains a public health crisis in Bangladesh. As arsenic has been shown to bind to human hemoglobin (Hb), hematologic mechanisms may play a role in the pathway through which arsenic exerts its toxicity. Two separate studies, a case-control and a cohort, were conducted to investigate the role of Hb in the development of arsenic-induced skin lesions. In the first, conditional logistic regression was used to investigate the effect of Hb on skin lesions among 900 case-control pairs from Pabna, Bangladesh, in which individuals were matched on gender, age, and location. In the second, mixed linear regression models were used to examine the association between toenail arsenic, urinary arsenic, and Hb within a cohort of 184 individuals from 50 families in the same region who did not have arsenic-induced skin lesions. Hb was significantly associated with skin lesions but this association was gender specific. In males, a 40% reduction in the odds of skin lesions occurred for every 1 g/dL increase in Hb (odds ratio, 0.60; 95% confidence interval, 0.49-0.73). No effect was observed for females (odds ratio, 1.16; 95% confidence interval, 0.92-1.46). In the cohort of 184 individuals, no associations between toenail arsenic or urinary arsenic species and Hb levels were observed. Low Hb levels may exacerbate the detrimental health effects of chronic arsenic poisoning. Whereas providing clean water remains the optimal solution to Bangladesh's problem of arsenic poisoning, improving nutrition and reducing iron-deficiency anemia may ameliorate negative health effects, such as skin lesions in individuals who have been exposed. PMID- 16702369 TI - Human papillomavirus testing following loop electrosurgical excision procedure identifies women at risk for posttreatment cervical intraepithelial neoplasia grade 2 or 3 disease. AB - BACKGROUND: Loop electrosurgical excision procedure (LEEP) is the predominant treatment for cervical intraepithelial neoplasia grade 2 or 3 (CIN2+) in the United States, yet following treatment approximately 10% of women are diagnosed again with CIN2+, necessitating close follow-up of such patients. METHODS: Surveillance strategies using cytology and/or human papillomavirus (HPV) testing were compared among women who underwent LEEP (n = 610) in the Atypical Squamous Cells of Undetermined Significance (ASCUS) Low-Grade Squamous Intraepithelial Lesion (LSIL) Triage Study. Cervical specimens, collected at 6-month visits for 2 years, were used for cytology, Hybrid Capture 2 (HC2) detection of carcinogenic HPVs, and PCR for genotyping of carcinogenic and noncarcinogenic HPV types. At exit, women had colposcopy for safety and disease ascertainment. RESULTS: At the visit post-LEEP (median time: 4.5 months after LEEP), 36.9% [95% confidence interval (95% CI), 32.7-41.1%] of women were positive for carcinogenic HPV by PCR and 33.7% (95% CI, 29.7-37.9) had ASCUS or more severe (ASCUS+) cytology. The overall 2-year cumulative incidence of histologically confirmed posttreatment CIN2+ was 7.0%; this could be further stratified by the HPV risk category detected at the 6-month visit after LEEP. The 2-year risk associated with HPV16 positivity was 37.0%, significantly higher than for other carcinogenic HPV types (10.8%, P < 0.001), noncarcinogenic types (1.5%, P < 0.001), or testing HPV negative (0%). Post-LEEP cytology (using a positive threshold of ASCUS+) was 78.1% (95% CI, 60.0-90.7%) sensitive for detection of posttreatment CIN2+. By comparison, PCR for carcinogenic HPV and combination testing (using a positive result from carcinogenic HPV testing or cytology as the test threshold with HPV negative ASCUS not referred) were significantly more sensitive (96.9% for each, P = 0.03); HC2 alone was nonsignificantly more sensitive (90.6%, P = 0.3). Specificity was similar for ASCUS+ cytology (69.1%, 95% CI, 64.6-73.3%) and PCR for carcinogenic HPV (67.1%, P = 0.5), yet was lower for HC2 (63.8%, P = 0.048) and combination testing (62.9%, P = 0.02). CONCLUSION: Women who tested positive after LEEP for carcinogenic HPV types, especially HPV16, had high risk of subsequent CIN2+. HPV-based detection methods, alone or in combination with cytology, may be useful to incorporate in post-LEEP management strategies. PMID- 16702370 TI - Smokers at higher risk for undetected antibody for oncogenic human papillomavirus type 16 infection. AB - OBJECTIVE: To determine the association between tobacco smoking and serologic evidence of human papillomavirus type 16 (HPV16)-specific antibodies among HPV16 DNA-positive women. DESIGN, SETTING, AND PARTICIPANTS: Baseline health history, physical examination, and laboratory data for 205 HPV16 DNA-positive women with no prior cytologic evidence of squamous intraepithelial lesions who were enrolled subsequently in a randomized clinical trial. MAIN OUTCOME MEASURE: HPV16-L1 antibody (anti-HPV16 antibody) detected from serum using RIA or ELISA. RESULTS: Eighty-seven percent (179 of 205) of women tested positive for HPV16 DNA using cervicovaginal swabs or lavage specimens, and 26 women showed similar results using swab specimens of external genitalia alone. HPV16-infected women who reported increasingly greater levels of daily cigarette smoking were less likely to test positive for anti-HPV16 antibodies than nonsmoking women (P = 0.02). Smokers were twice as likely as nonsmokers to test negative for anti-HPV16 antibodies, even after controlling for the effects of other covariates in the analyses (adjusted odds ratio, 0.5; 95% confidence limits, 0.2-0.9). Although Papanicolaou test findings and smoking characteristics were poorly correlated (r(2) = 0.01), women who showed atypical cells of unknown significance or squamous intraepithelial lesion were twice as likely to test anti-HPV16 antibody positive as women who showed normal Papanicolaou tests (adjusted odds ratio, 2.0; 95% confidence limits, 1.1-3.7). CONCLUSION: These data suggest that smoking may influence the long-term risk for cancer by perturbing early immune responses to the virus and may increase the likelihood of persistent infection. Patient education messages should alert women to this additional risk of smoking. A clinical trial of smoking cessation should be explored as a therapeutic intervention for primary HPV16 infection. PMID- 16702372 TI - Associations of classic Kaposi sarcoma with common variants in genes that modulate host immunity. AB - Classic Kaposi sarcoma (CKS) is an inflammatory-mediated neoplasm primarily caused by Kaposi sarcoma-associated herpesvirus (KSHV). Kaposi sarcoma lesions are characterized, in part, by the presence of proinflammatory cytokines and growth factors thought to regulate KSHV replication and CKS pathogenesis. Using genomic DNA extracted from 133 CKS cases and 172 KSHV-latent nuclear antigen positive, population-based controls in Italy without HIV infection, we examined the risk of CKS associated with 28 common genetic variants in 14 immune modulating genes. Haplotypes were estimated for IL1A, IL1B, IL4, IL8, IL8RB, IL10, IL12A, IL13, and TNF. Compared with controls, CKS risk was decreased with 1235T/-1010G-containing diplotypes of IL8RB (odds ratio, 0.49; 95% confidence interval, 0.30-0.78; P = 0.003), whereas risk was increased with diplotypes of IL13 containing the promoter region variant 98A (rs20541, alias +130; odds ratio, 1.88; 95% confidence interval, 1.15-3.08; P = 0.01) when considered in multivariate analysis. Risk estimates did not substantially vary by age, sex, incident disease, or disease burden. Our data provide preliminary evidence for variants in immune-modulating genes that could influence the risk of CKS. Among KSHV-seropositive Italians, CKS risk was associated with diplotypes of IL8RB and IL13, supporting laboratory evidence of immune-mediated pathogenesis. PMID- 16702371 TI - Statin use and risk of lymphoid neoplasms: results from the European Case-Control Study EPILYMPH. AB - BACKGROUND: Statins, drugs used to treat dyslipidemia, may have anticancer properties. We have evaluated lymphoma risk associated with regular statin use in an international case-control study. METHODS: This case-control study included 2,362 cases of incident B- and T-cell lymphoma from Czech Republic, France, Germany, Ireland, Italy, and Spain and 2,206 hospital or population controls. Information on drug use, diagnosis at admission (for hospital controls), and putative risk factors for lymphoma was collected with personal interviews. Hospital controls admitted for diseases possibly entailing use of statins were excluded from the analysis. RESULTS: The odds ratio for regular statin use was 0.61 (95% confidence interval, 0.45-0.84); all major lymphoma subtypes showed similarly decreased risks. Decreased risks were observed in all centers. Duration of statin use was not associated with a greater reduction in the risk of lymphoma. Use of other lipid lowering drugs, such as fibrates, did not significantly modify the risk of lymphoma (odds ratio, 0.75; 95% confidence interval, 0.44-1.27). CONCLUSION: Statin use was associated with an important reduction in lymphoma risk, adding to the growing evidence of anticancer properties of this group of drugs. These results are reassuring for the increasing number of patients taking statins on a regular basis. PMID- 16702373 TI - Role of the Nijmegen breakage syndrome 1 gene in familial and sporadic prostate cancer. AB - The Nijmegen breakage syndrome 1 (NBS1) gene, which participates in DNA double strand break repair, has been postulated to be a susceptibility factor for a number of cancers, including prostate cancer. Numerous mutations have been identified in NBS1, including the founder mutation 657del5. In this study, a number of analyses were done to determine whether mutations in NBS1 are associated with an increased risk for prostate cancer. The frequency of the 657del5 mutation in both familial prostate cancer cases (1,819 affected men among 909 families) and sporadic prostate cancer cases (1,218 affected men) collected from five centers participating in the International Consortium for Prostate Cancer Genetics were compared with that found in 697 normal controls. Seven individuals were identified to carry the mutation among the 3,037 cases screened: four in the familial group (three from one family and one from another) and three in the sporadic cases. The carrier frequency was 0.22% (2 of 909) for the probands and 0.25% (3 of 1,218) for the sporadic cases of prostate cancer. The 657del5 mutation was not detected in either the 293 unaffected members of the prostate cancer families or in the 697 control samples tested. The entire NBS1 gene was also sequenced in 20 of the youngest affected individuals from the Finnish group of familial cases to identify the presence of possible mutations in this high-risk group. One rare (D95N) and one common (E185Q) missense alteration was identified. More detailed analyses of the E185Q polymorphism, along with a third rare variant (R215W), failed to show an association with prostate cancer. Because the 657del5 mutation was absent from the control population, we are unable to determine if this alteration predisposes to prostate cancer. However, our data does suggest that mutations within NBS1, and in particular, 657del5, do not significantly contribute to the overall prostate cancer burden within our patient samples. PMID- 16702374 TI - Plasma antibodies against Trichomonas vaginalis and subsequent risk of prostate cancer. AB - BACKGROUND: Although several previous case-control studies have investigated associations between sexually transmitted infections (STI) and prostate cancer, most have focused on gonorrhea and syphilis, two well-recognized, symptomatic STIs. Another STI of interest for prostate carcinogenesis is trichomonosis, a less well recognized and frequently asymptomatic STI with known prostate involvement. We investigated this infection in relation to incident prostate cancer in a nested case-control study within the Health Professionals Follow-up Study. METHODS: Prostate cancer cases were men diagnosed with prostate cancer between the date of blood draw (1993-1995) and 2000 (n = 691). Controls were men who had had at least one prostate-specific antigen test and who were free of prostate cancer and alive at the time of case diagnosis. One control was individually matched to each case by age (n = 691). Serologic evidence of a history of trichomonosis was assessed by a recombinant Trichomonas vaginalis alpha-actinin IgG ELISA. RESULTS: Thirteen percent of cases and 9% of controls were seropositive for trichomonosis (adjusted odds ratio, 1.43; 95% confidence interval, 1.00-2.03). This association persisted after additional adjustment for such factors as a history of other STIs, and was strongest among men who used aspirin infrequently over the course of their lives (odds ratio, 2.05; 95% confidence interval, 1.05-4.02, P(interaction) = 0.11). CONCLUSIONS: Serologic evidence of a history of trichomonosis was positively associated with incident prostate cancer in this large, nested case-control study of male health professionals. As this study is the first, to our knowledge, to investigate associations between T. vaginalis serology and prostate cancer, additional studies are necessary before conclusions can be made. PMID- 16702375 TI - Relationship between established breast cancer risk factors and risk of seven different histologic types of invasive breast cancer. AB - BACKGROUND: Important differences in the contributions of certain exposures to the risks of ductal versus lobular breast carcinomas have been observed, but few studies have evaluated the relationships between established breast cancer risk factors and other histologic types. METHODS: Information on family history of cancer and reproductive, hormonal, anthropometric, and lifestyle characteristics were collected in a multicenter population-based case-control study consisting of 3,463 ductal, 274 lobular, 261 ductal-lobular, 91 medullary, 77 tubular, 70 comedo, and 61 mucinous invasive breast carcinoma cases (ages 35-64 years, newly diagnosed 1994-1998) and 4,682 controls. Associations between each of these histologic types and various exposures were evaluated using polytomous regression. RESULTS: Heterogeneity in the risks of different histologic types of breast cancer was observed for three exposures: menopausal hormone use, body mass index (BMI), and alcohol consumption. Specifically, current use of unopposed estrogen was associated with a reduced risk of ductal carcinoma and increased risk of comedocarcinoma, and current use of estrogen and progestin was associated with elevated risks of ductal-lobular and tubular carcinomas. Among postmenopausal women, BMI was only inversely related to risk of ductal-lobular carcinoma, and alcohol use was only positively related to risk of lobular carcinoma. CONCLUSIONS: Variations in the associations between known breast cancer risk factors and risk of different breast cancer histologies were observed. Although these findings require confirmation, and the analyses of some histologic groups were limited by small sample sizes, they provide some insight into the different etiologies of various histologic subtypes of breast cancer. PMID- 16702376 TI - Value for the future and breast cancer-preventive health behavior. AB - BACKGROUND: Time preference, or the extent to which people discount future benefits in favor of immediate benefits, might represent an important determinant of preventive health behavior, but the little research thus far on this association has yielded mixed results. This study examined the association between future time preference and use of genetic counseling for BRCA1/2 testing and how this association may differ from the relationship between future time preference and mammography screening and self-breast examination. EXPERIMENTAL DESIGN: A health system-based case-control study with a nested cross-sectional survey. Eight hundred women who saw a primary care physician in the University of Pennsylvania Health System in the 3 years before the study, of whom 234 had undergone BRCA1/2 counseling (cases) and of whom 566 had not (controls). RESULTS: Placing a relatively greater value on future benefits than present benefits was strongly associated with use of BRCA1/2 counseling [odds ratio (OR), 3.0 for one point increase in future time preference; 95% confidence intervals (CI), 1.9 4.9]. Future time preference was weakly associated with adherence to annual mammography (OR, 1.3; 95% CI, 0.81-2.2), and was not associated with monthly self breast examination (OR, 1.03; 95% CI, 0.75-1.4). A stronger future orientation was seen in women who had higher levels of education (P = 0.0021) or income (P = 0.0011). CONCLUSION: Time preference is strongly associated with use of BRCA1/2 counseling. Time preference is more weakly associated with mammography adherence and is not associated with breast self-examination. This variation may reflect the degree to which the behavior is seen as related to future risk. PMID- 16702377 TI - Mammographic density as a surrogate marker for the effects of hormone therapy on risk of breast cancer. AB - BACKGROUND: Some types of hormone therapy increase both risk of breast cancer and mammographic density, a risk factor for the disease, suggesting that mammographic density may be a surrogate marker for the effects of hormones on risk of breast cancer. This research was undertaken to determine whether the effect of hormone therapy on breast cancer risk is mediated by its effect on mammographic density. METHODS: Individually matched cases and controls from three nested case-control studies in breast screening populations were studied. Cases had developed invasive breast cancer at least 12 months after the initial screen. Information was collected on hormone use and other risk factors at the time of the baseline mammogram, and percent density was measured by a computer-assisted method. RESULTS: There were 1,748 postmenopausal women, of whom 426 (24.4%) were using hormones at the time of their initial screening mammogram. Current use of hormone therapy was associated with an increased risk of breast cancer (odds ratio, 1.26; 95% confidence interval, 1.0-1.6) that was little changed by adjustment for percent density in the baseline mammogram (odds ratio, 1.19; 95% confidence interval, 0.9-1.5). Percent density in the baseline mammogram was among cases greater in current users of hormones that in never-users (difference = 5.0%, P < 0.001), but the difference was smaller and nonsignificant in controls (difference = 1.6%, P = 0.3). CONCLUSION: Although the effects of hormone therapy on mammographic density were greater in cases than controls, we did not find evidence that these effects were causally related to risk of breast cancer. PMID- 16702378 TI - The association of plasma DHEA and DHEA sulfate with breast cancer risk in predominantly premenopausal women. AB - Concentrations of adrenal androgens are positively associated with postmenopausal breast cancer risk; however, results in premenopausal women are conflicting. Therefore, we conducted a prospective nested case-control study within the Nurses' Health Study II cohort to examine the relationship of DHEA and DHEA sulfate (DHEAS) with breast cancer risk in predominantly premenopausal women. Blood samples were collected from 1996 to 1999. The analysis included 317 cases of breast cancer diagnosed after blood collection and before June 1, 2003; for each case, two controls were matched on age, fasting status, time of day and month of blood collection, race/ethnicity, and timing of blood draw within the menstrual cycle. No associations were observed between DHEA or DHEAS levels and breast cancer risk overall [in situ and invasive; DHEA relative risk (RR), top versus bottom quartile, 1.2; 95% confidence interval (95% CI), 0.8-1.8, P(trend) = 0.53; DHEAS RR, 1.3; 95% CI, 0.9-2.0; P(trend) = 0.07]. However, both DHEA and DHEAS were positively associated with estrogen receptor-positive/progesterone receptor-positive breast cancer (DHEA RR, 1.6; 95% CI, 0.9-2.8, P(trend) = 0.09; DHEAS RR, 1.9; 95% CI, 1.1-3.3, P(trend) = 0.02). We observed a significant interaction by age, with an RR for DHEAS of 0.8 (95% CI, 0.4-1.5, P(trend) = 0.62) for women <45 years old and 2.0 (95% CI, 1.2-3.5, P(trend) = 0.003) for women >/=45 years old; results were similar for DHEA. Our results suggest that adrenal androgens are positively associated with breast cancer among predominately premenopausal women, especially for estrogen receptor positive/progesterone receptor-positive tumors and among women over age 45 years. PMID- 16702379 TI - Reproducibility of plasma steroid hormones, prolactin, and insulin-like growth factor levels among premenopausal women over a 2- to 3-year period. AB - Few studies have evaluated whether a single blood hormone measurement, as is available in most epidemiologic studies, sufficiently characterizes a premenopausal woman's long-term hormone levels; there is particular concern whether sex steroid hormones, which fluctuate during the menstrual cycle, are reliable. We conducted a prospective study within the Nurses' Health Study II to examine the reproducibility of plasma estrogens, androgens, progesterone, prolactin, sex hormone binding globulin, insulin-like growth factor-I (IGF-I), and IGF binding protein-3 (IGFBP-3). One blood sample per year over 3 years was collected from 113 premenopausal women during both the follicular and luteal phases of the menstrual cycle. We calculated intraclass correlation coefficients (ICC) across the three samples for all women. Among estrogens, ICCs ranged from 0.38 (estradiol) to 0.60 (estrone sulfate) in the follicular phase and from 0.44 (estrone) to 0.69 (estrone sulfate) in the luteal phase. Among androgens, ICCs ranged from 0.58 (androstenedione) to 0.94 [dehydroepiandrostenedione sulfate (DHEAS)] in the follicular phase and from 0.56 (testosterone) to 0.81 (DHEAS) in the luteal phase. When values were averaged across the follicular and luteal phases, the ICC for prolactin was 0.64 whereas ICCs for IGF-I and IGFBP-3 were 0.86 and 0.82, respectively. The ICC for progesterone in the luteal phase was only 0.29. These data suggest that for androgens, estrone sulfate, prolactin, IGF I, and IGFBP-3, a single measurement can reliably categorize average levels over at least a 3-year period in premenopausal women. For estrone and estradiol, where ICCs were relatively low, it is important to use reproducibility data such as those to correct for measurement error in epidemiologic studies. PMID- 16702380 TI - NAD(P)H:quinone oxidoreductase 1 (NQO1) Pro187Ser polymorphism and the risk of lung, bladder, and colorectal cancers: a meta-analysis. AB - NAD(P)H:quinone oxidoreductase 1 (NQO1) is a cytosolic enzyme that catalyzes the two-electron reduction of quinoid compounds into hydroquinones, their less toxic form. A sequence variant at position 609 (C --> T) in the NQO1 gene encodes an enzyme with reduced quinone reductase activity in vitro and thus was hypothesized to affect cancer susceptibility. We conducted meta-analyses focusing on three cancer sites (lung, bladder, and colorectum) to summarize the findings from the current literature and to explore sources of heterogeneity. RESULTS: There is no clear association between the NQO1 Pro187Ser polymorphism and lung cancer risk in the three ethnic groups examined: odds ratio (OR(White)) C/T + T/T versus C/C = 1.04 [95% confidence interval (95% CI), 0.96-1.13], OR(Asian) = 0.99 (95% CI, 0.72-1.34), and OR(Blacks) = 0.95 (95% CI, 0.66-1.36). However, a modestly increased risk was suggested for the variant homozygotes in whites (OR T/T versus C/C, 1.19; 95% CI, 0.94-1.50). Analysis excluding one outlier study suggested the variant allele may be associated with reduced lung cancer risk in Asians. Meta analyses for bladder and colorectal cancer suggested a statistically significant association with the variant genotypes in whites. In stratified analyses, the NQO1 Pro187Ser variant genotypes were associated with slightly increased lung cancer risk in white ever smokers but not in white never smokers and were mainly associated with a reduced risk of lung adenocarcinoma but not squamous cell carcinoma in Asians. CONCLUSIONS: Results from our meta-analyses suggest that the variant NQO1 Pro187Ser genotype may affect individual susceptibility to lung, bladder, and colorectal cancer. Such effects of the NQO1 polymorphism seem to be modified by ethnicity and smoking status. PMID- 16702381 TI - 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol and its glucuronides in the urine of infants exposed to environmental tobacco smoke. AB - Biomarkers of carcinogen uptake could provide important information pertinent to the question of exposure to environmental tobacco smoke (ETS) in childhood and cancer development later in life. Previous studies have focused on exposures before birth and during childhood, but carcinogen uptake from ETS in infants has not been reported. Exposures in infants could be higher than in children or adults because of their proximity to parents who smoke. Therefore, we quantified 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol and its glucuronides (total NNAL) in the urine of 144 infants, ages 3 to 12 months, who lived in homes with parents who smoked. Total NNAL is an accepted biomarker of uptake of the tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone. Cotinine and its glucuronide (total cotinine) and nicotine and its glucuronide (total nicotine) were also quantified. Total NNAL was detectable in 67 of 144 infants (46.5%). Mean levels of total NNAL in the 144 infants were 0.083 +/- 0.200 pmol/mL, whereas those of total cotinine and total nicotine were 0.133 +/- 0.190 and 0.069 +/- 0.102 nmol/mL, respectively. The number of cigarettes smoked per week in the home or car by any family member when the infant was present was significantly higher (P < 0.0001) when NNAL was detected than when it was not (76.0 +/- 88.1 versus 27.1 +/- 38.2). The mean level of NNAL detected in the urine of these infants was higher than in most other field studies of ETS exposure. The results of this study show substantial uptake of 4-(methylnitrosamino)-1-(3-pyridyl)-1 butanone in infants exposed to ETS and support the concept that persistent ETS exposure in childhood could be related to cancer later in life. PMID- 16702382 TI - A comparison of carotenoids, retinoids, and tocopherols in the serum and buccal mucosa of chronic cigarette smokers versus nonsmokers. AB - BACKGROUND: Cigarette smoking, a major risk factor for oropharyngeal cancer, is reported to alter oral levels of carotenoids and tocopherols. Such effects may be important because these nutrients, as well as retinoids, are putative chemoprotective agents. OBJECTIVES: To determine whether chronic smoking is associated with altered concentrations of these nutrients in serum and buccal mucosa; to distinguish whether such effects are ascribable to diet; and to determine whether oral concentrations of these nutrients correlate with a putative biomarker of oral cancer risk. METHODS: Serum and buccal mucosal cells (BMC) were analyzed for these nutrients and for BMC micronuclei in smokers (n = 35) and nonsmokers (n = 21). RESULTS: General linear regression with adjustments for dietary intake showed that smokers possess lower serum concentrations of beta and alpha-carotene, cryptoxanthin, lutein, and zeaxanthin (P /=5% frequency in at least one ethnic group) spanning the PTEN gene to define the common alleles in these populations. These SNPs were in strong linkage disequilibrium, indicating that our survey captured most of the common sequence variation across this locus. Eight tagging SNPs were selected to predict the common PTEN haplotypes (>/=0.05 frequency) in these populations (two additional tagging SNPs were required for African Americans). These SNPs were evaluated in a breast cancer case-control study (cases, n = 1,615; controls, n = 1,962) and prostate cancer case-control study (cases, n = 2,320; controls, n = 2,290) nested within the Multiethnic Cohort Study. Multiple testing was explicitly accounted for by applying a permutation based framework. We found no strong association with any common haplotype in relation to breast or prostate cancer risk. In summary, our results show that common variants in PTEN do not substantially influence risk of these two common cancers. PMID- 16702387 TI - The influence of statin use on breast density. AB - OBJECTIVE: To evaluate if 3-hydroxy-3-methylglutaryl CoA reductase reductase inhibitor use (statins) alters mammography measured breast density. METHODS: Cohort study of women ages 50 to 80 years with two mammography screenings. Changes in BI-RADS breast density between screenings was compared for nonusers, initiators, discontinuers, continuers, and any users of statins. RESULTS: Statin use was not associated with increases or decreases in breast density compared with nonusers after adjusting for age, body mass index, change in body mass index, hormone therapy use, and time between screenings. Cumulative days of statin use during the year before screening was not associated with changes in breast density in any of the groups of statin users. When hormone therapy users were excluded, any statin use was associated with an increase in breast density compared with nonusers (odds ratio, 1.2; 95% confidence interval, 1.0-1.5). CONCLUSIONS: We found no association between statin use and change in breast density in general, but statin use may be associated with increases in breast density among nonusers of hormone therapy. PMID- 16702389 TI - A reproducibility study for a fluoride assay in bone. AB - Osteosarcoma is a rare malignancy of largely unknown etiology. Although there is no consistent evidence for an association between fluoridation and cancer, some concerns remain about osteosarcoma. As part of the design of a collaborative study, bone samples were collected to allow for an evaluation of the association between osteosarcoma risk and individual fluoride exposure measured by levels of fluoride in bone. In this report, we provide the results of pilot experiments to consider issues that arose during the study design and to assess the reliability of the bone assays. Correlations of fluoride levels between normal bone near the affected area and iliac crest bone were strong and positive. The day-to-day laboratory analysis of fluoride in human and deer jaw bone yielded acceptable average coefficients of variation below 10% and an overall estimate of 5%. The intraclass correlation (ICC) is of particular importance to epidemiologists because it indicates the effect of measurement error on study results. Here, the estimated ICC is 0.86, and the estimated downward bias is only 14%. Hence, the ICC is strong enough so that the estimates of the relative risk will suffer little attenuation from lab measurements. PMID- 16702388 TI - A phase II study with antioxidants, both in the diet and supplemented, pharmaconutritional support, progestagen, and anti-cyclooxygenase-2 showing efficacy and safety in patients with cancer-related anorexia/cachexia and oxidative stress. AB - PURPOSE: To test the efficacy and safety of an integrated treatment based on a pharmaconutritional support, antioxidants, and drugs, all given orally, in a population of advanced cancer patients with cancer-related anorexia/cachexia and oxidative stress. PATIENTS AND METHODS: An open early-phase II study was designed according to the Simon two-stage design. The integrated treatment consisted of diet with high polyphenols content (400 mg), antioxidant treatment (300 mg/d alpha-lipoic acid + 2.7 g/d carbocysteine lysine salt + 400 mg/d vitamin E + 30,000 IU/d vitamin A + 500 mg/d vitamin C), and pharmaconutritional support enriched with 2 cans per day (n-3)-PUFA (eicosapentaenoic acid and docosahexaenoic acid), 500 mg/d medroxyprogesterone acetate, and 200 mg/d selective cyclooxygenase-2 inhibitor celecoxib. The treatment duration was 4 months. The following variables were evaluated: (a) clinical (Eastern Cooperative Oncology Group performance status); (b) nutritional [lean body mass (LBM), appetite, and resting energy expenditure]; (c) laboratory [proinflammatory cytokines and leptin, reactive oxygen species (ROS) and antioxidant enzymes]; (d) quality of life (European Organization for Research and Treatment of Cancer QLQ C30, Euro QL-5D, and MFSI-SF). RESULTS: From July 2002 to January 2005, 44 patients were enrolled. Of these, 39 completed the treatment and were assessable. Body weight increased significantly from baseline as did LBM and appetite. There was an important decrease of proinflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor-alpha, and a negative relationship worthy of note was only found between LBM and IL-6 changes. As for quality of life evaluation, there was a marked improvement in the European Organization for Research and Treatment of Cancer QLQ-C30, Euro QL-5D(VAS), and multidimensional fatigue symptom inventory short form scores. At the end of the study, 22 of the 39 patients were "responders" or "high responders." The minimum required was 21; therefore, the treatment was effective and more importantly was shown to be safe. CONCLUSION: The efficacy and safety of the treatment have been shown by the study; therefore, a randomized phase III study is warranted. PMID- 16702390 TI - The effects of GSTM1 and GSTT1 polymorphisms on micronucleus frequencies in human lymphocytes in vivo. AB - The influence of genetic polymorphisms in GSTM1 and GSTT1 genes on micronucleus frequencies in human peripheral blood lymphocytes was assessed through a pooled analysis of data from seven laboratories that did biomonitoring studies using the in vivo cytokinesis-block micronucleus assay. A total of 301 nonoccupationally exposed individuals (207 males and 94 females) and 343 workers (237 males and 106 females) occupationally exposed to known or suspected genotoxic substances were analyzed by Poisson regression. The results of the pooled analysis indicate that the GSTT1 null subjects had lower micronucleus frequencies than their positive counterparts in the total population (frequency ratio, 0.55; 95% confidence interval, 0.33-0.89). The protective effect of this genotype is reversed with increasing age, with a frequency ratio of 1.33 (95% confidence interval, 1.06 1.68) in subjects aged 60 years. A significant overall increase in micronucleus frequency with age and gender (P < 0.001 and P = 0.024, respectively) was observed, females having higher micronucleus frequencies than males, when occupationally exposed (P = 0.002). Nonoccupationally exposed smokers had lower micronucleus frequencies than nonsmokers (P = 0.001), whereas no significant difference in micronucleus level was observed between smokers and nonsmokers in the occupationally exposed group (P = 0.79). This study confirms that pooled analyses, by increasing the statistical power, are adequate for assessing the involvement of genetic variants on genome stability and for resolving discrepancies among individual studies. PMID- 16702391 TI - No association between polymorphisms in CYP2E1, GSTM1, NAT1, NAT2 and the risk of gastric adenocarcinoma in the European prospective investigation into cancer and nutrition. PMID- 16702392 TI - Selenomethionine treatment does not alter gene expression in normal squamous esophageal mucosa in a high-risk Chinese population. AB - Selenium is a promising cancer chemoprevention agent. A recent randomized controlled chemoprevention trial found that selenomethionine (SeMet) supplementation for 10 months favorably effected a change in esophageal dysplasia grade among participants who started the trial with mild dysplasia. To further explore the role of SeMet in this trial, we compared gene expression profiles by treatment group using Affymetrix HU 133A chips in before/after supplementation paired normal esophageal biopsies from a subset of 29 trial participants, 16 who received SeMet, and 13 who received placebo. Using P < 0.001 as a cutoff, 11 differentially expressed genes were found in the SeMet supplementation group but these genes did not include either known selenoprotein genes or genes previously shown to be modulated by selenium treatment. Because the number of differentially expressed genes (n = 11) was less than expected by chance (n = 18), we concluded that SeMet supplementation had no measurable effect on gene expression in the normal squamous esophagus of these subjects with dysplasia. PMID- 16702393 TI - Genetic polymorphisms in the DNA double-strand break repair genes XRCC3, XRCC2, and NBS1 are not associated with acute side effects of radiotherapy in breast cancer patients. PMID- 16702394 TI - Dietary factors and cancers of the renal pelvis and ureter. PMID- 16702395 TI - Is there a right-sided shift for colorectal cancer in women compared with men? PMID- 16702396 TI - Food frequency questionnaires: not irrelevant yet. PMID- 16702397 TI - Imprinting mechanisms--it only takes two. PMID- 16702398 TI - Nucleolar dominance: a model for rRNA gene silencing. PMID- 16702399 TI - Fertility versus disease resistance, a hard choice. PMID- 16702400 TI - Genetics and biology of pancreatic ductal adenocarcinoma. AB - Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of cancer death in the United States with a median survival of <6 mo and a dismal 5-yr survival rate of 3%-5%. The cancer's lethal nature stems from its propensity to rapidly disseminate to the lymphatic system and distant organs. This aggressive biology and resistance to conventional and targeted therapeutic agents leads to a typical clinical presentation of incurable disease at the time of diagnosis. The well-defined serial histopathologic picture and accompanying molecular profiles of PDAC and its precursor lesions have provided the framework for emerging basic and translational research. Recent advances include insights into the cancer's cellular origins, high-resolution genomic profiles pointing to potential new therapeutic targets, and refined mouse models reflecting both the genetics and histopathologic evolution of human PDAC. This confluence of developments offers the opportunity for accelerated discovery and the future promise of improved treatment. PMID- 16702401 TI - ASPP2 is a haploinsufficient tumor suppressor that cooperates with p53 to suppress tumor growth. AB - ASPP2 stimulates the apoptotic function of the p53 family in vivo. We show here that ASPP2-/- pups died before weaning. This postnatal lethality was significantly enhanced in p53+/- background and both deletions are synthetic lethal. ASPP2+/- mice developed spontaneous tumors. The tumor onset was accelerated by gamma-irradiation or in p53+/- background. Tumors derived from ASPP2+/- mice retained wild-type ASPP2 allele even though some of them lost p53. These provide the first genetic evidence that ASPP2 is a haploinsufficient tumor suppressor that shares overlapping function(s) with p53 in mouse development and tumor suppression. PMID- 16702402 TI - Elongation of the Kcnq1ot1 transcript is required for genomic imprinting of neighboring genes. AB - The imprinted gene cluster at the telomeric end of mouse chromosome 7 contains a differentially methylated CpG island, KvDMR, that is required for the imprinting of multiple genes, including the genes encoding the maternally expressed placental-specific transcription factor ASCL2, the cyclin-dependent kinase CDKN1C, and the potassium channel KCNQ1. The KvDMR, which maps within intron 10 of Kcnq1, contains the promoter for a paternally expressed, noncoding, antisense transcript, Kcnq1ot1. A 244-base-pair deletion of the promoter on the paternal allele leads to the derepression of all silent genes tested. To distinguish between the loss of silencing as the consequence of the absence of transcription or the transcript itself, we prematurely truncated the Kcnq1ot1 transcript by inserting a transcriptional stop signal downstream of the promoter. We show that the lack of a full-length Kcnq1ot1 transcript on the paternal chromosome leads to the expression of genes that are normally paternally repressed. Finally, we demonstrate that five highly conserved repeats residing at the 5' end of the Kcnq1ot1 transcript are not required for imprinting at this locus. PMID- 16702403 TI - Systematic identification and functional screens of uncharacterized proteins associated with eukaryotic ribosomal complexes. AB - Translation regulation is a critical means by which cells control growth, division, and apoptosis. To gain further insight into translation and related processes, we performed multifaceted mass spectrometry-based proteomic screens of yeast ribosomal complexes and discovered an association of 77 uncharacterized yeast proteins with ribosomes. Immunoblotting revealed an EDTA-dependent cosedimentation with ribosomes in sucrose gradients for 11 candidate translation machinery-associated (TMA) proteins. Tandem affinity purification linked one candidate, LSM12, to the RNA processing proteins PBP1 and PBP4. A second candidate, TMA46, interacted with RBG1, a GTPase that interacts with ribosomes. By adapting translation assays to high-throughput screening methods, we showed that null yeast strains harboring deletions for several of the TMA genes had alterations in protein synthesis rates (TMA7 and TMA19), susceptibility to drugs that inhibit translation (TMA7), translation fidelity (TMA20), and polyribosome profiles (TMA7, TMA19, and TMA20). TMA20 has significant sequence homology with the oncogene MCT-1. Expression of human MCT-1 in the Deltatma20 yeast mutant complemented translation-related defects, strongly implying that MCT-1 functions in translation-related processes. Together these findings implicate the TMA proteins and, potentially, their human homologs, in translation related processes. PMID- 16702404 TI - Nuclear receptor TLX prevents retinal dystrophy and recruits the corepressor atrophin1. AB - During mammalian embryogenesis, precise coordination of progenitor cell proliferation and differentiation is essential for proper organ size and function. The involvement of TLX (NR2E1), an orphan nuclear receptor, has been implicated in ocular development, as Tlx-/- mice exhibit visual impairment. Using genetic and biochemical approaches, we show that TLX modulates retinal progenitor cell proliferation and cell cycle re-entry by directly regulating the expression of Pten and its target cyclin D1. Additionally, TLX finely tunes the progenitor differentiation program by modulating the phospholipase C and mitogen-activated protein kinase (MAPK) pathways and the expression of an array of cell type specific transcriptional regulators. Consequently, Tlx-/- mice have a dramatic reduction in retina thickness and enhanced generation of S-cones, and develop severe early onset retinal dystrophy. Furthermore, TLX interacts with atrophin1 (Atn1), a corepressor that is involved in human neurodegenerative dentatorubral pallidoluysian atrophy (DRPLA) and that is essential for development of multiple tissues. Together, these results reveal a molecular strategy by which an orphan nuclear receptor can precisely orchestrate tissue-specific proliferation and differentiation programs to prevent retinal malformation and degeneration. PMID- 16702405 TI - MOZ is essential for maintenance of hematopoietic stem cells. AB - Monocytic leukemia zinc-finger protein (MOZ), a MYST family histone acetyltransferase, is involved in the chromosome translocations associated with acute myeloid leukemia. MOZ acts as a transcriptional coactivator for AML1, which is essential for establishment of definitive hematopoiesis. To investigate the roles of MOZ in normal hematopoiesis, we generated MOZ-null mice. MOZ-/- mice died around embryonic day 15 (E15). In MOZ-/- E14.5 embryos, hematopoietic stem cells, lineage-committed progenitors, and B lineage cells were severely reduced. On the other hand, arrest of erythroid maturation and elevated myeloid lineage populations were observed. MOZ-deficient fetal liver cells could not reconstitute hematopoiesis of recipients after transplantation. Analysis using microarray and flow cytometry revealed that expression of thrombopoietin receptor (c-Mpl), HoxA9, and c-Kit was down-regulated. These results show that MOZ is required for maintenance of hematopoietic stem cells, and that it plays a role in differentiation of erythroid and myeloid cells. Some aspects of the MOZ-/- phenotype are similar to that observed in PU.1-deficient mice. MOZ was able to interact with PU.1 and activate PU.1-dependent transcription, thus suggesting a physical and functional link between PU.1 and MOZ. PMID- 16702406 TI - Loss of transforming growth factor-beta type II receptor promotes metastatic head and-neck squamous cell carcinoma. AB - The prognosis of head-and-neck squamous cell carcinoma (HNSCC) has not been improved in the past 20 years. Validation of HNSCC biomarkers for targeted therapy has been hindered by a lack of animal models mimicking human HNSCC at both the pathological and molecular levels. Here we report that overexpression of K-ras or H-ras and loss of transforming growth factor-beta type II receptor (TGFbetaRII) are common events in human HNSCC. Activation of either K-ras or H ras in combination with TGFbetaRII deletion from mouse head-and-neck epithelia caused HNSCC with complete penetrance, some of which progressed to metastases. These tumors displayed pathology indistinguishable from human HNSCCs and exhibited multiple molecular alterations commonly found in human HNSCCs. Additionally, elevated endogenous TGFbeta1 in these lesions contributed to inflammation and angiogenesis. Our data suggest that targeting common oncogenic pathways in tumor epithelia together with blocking the effect of TGFbeta1 on tumor stroma may provide a novel therapeutic strategy for HNSCC. PMID- 16702407 TI - DNA damage triggers nucleotide excision repair-dependent monoubiquitylation of histone H2A. AB - Chromatin changes within the context of DNA repair remain largely obscure. Here we show that DNA damage induces monoubiquitylation of histone H2A in the vicinity of DNA lesions. Ultraviolet (UV)-induced monoubiquitylation of H2A is dependent on functional nucleotide excision repair and occurs after incision of the damaged strand. The ubiquitin ligase Ring2 is required for the DNA damage-induced H2A ubiquitylation. UV-induced ubiquitylation of H2A is dependent on the DNA damage signaling kinase ATR (ATM- and Rad3-related) but not the related kinase ATM (ataxia telangiectasia-mutated). Although the response coincides with phosphorylation of variant histone H2AX, H2AX was not required for H2A ubiquitylation. Together our data show that monoubiquitylation of H2A forms part of the cellular response to UV damage and suggest a role of this modification in DNA repair-induced chromatin remodeling. PMID- 16702408 TI - Keratin 17 modulates hair follicle cycling in a TNFalpha-dependent fashion. AB - Mammalian hair follicles cycle between stages of rapid growth (anagen) and metabolic quiescence (telogen) throughout life. Transition from anagen to telogen involves an intermediate stage, catagen, consisting of a swift, apoptosis-driven involution of the lower half of the follicle. How catagen is coordinated, and spares the progenitor cells needed for anagen re-entry, is poorly understood. Keratin 17 (K17)-null mice develop alopecia in the first week post-birth, correlating with hair shaft fragility and untimely apoptosis in the hair bulb. Here we show that this abnormal apoptosis reflects premature entry into catagen. Of the proapoptotic challenges tested, K17-null skin keratinocytes in primary culture are selectively more sensitive to TNFalpha. K17 interacts with TNF receptor 1 (TNFR1)-associated death domain protein (TRADD), a death adaptor essential for TNFR1-dependent signal relay, suggesting a functional link between this keratin and TNFalpha signaling. The activity of NF-kappaB, a downstream target of TNFalpha, is increased in K17-null skin. We also find that TNFalpha is required for a timely anagen-catagen transition in mouse pelage follicles, and that its ablation partially rescues the hair cycling defect of K17-null mice. These findings identify K17 and TNFalpha as two novel and interdependent regulators of hair cycling. PMID- 16702409 TI - The chicken talpid3 gene encodes a novel protein essential for Hedgehog signaling. AB - Talpid3 is a classical chicken mutant with abnormal limb patterning and malformations in other regions of the embryo known to depend on Hedgehog signaling. We combined the ease of manipulating chicken embryos with emerging knowledge of the chicken genome to reveal directly the basis of defective Hedgehog signal transduction in talpid3 embryos and to identify the talpid3 gene. We show in several regions of the embryo that the talpid3 phenotype is completely ligand independent and demonstrate for the first time that talpid3 is absolutely required for the function of both Gli repressor and activator in the intracellular Hedgehog pathway. We map the talpid3 locus to chromosome 5 and find a frameshift mutation in a KIAA0586 ortholog (ENSGALG00000012025), a gene not previously attributed with any known function. We show a direct causal link between KIAA0586 and the mutant phenotype by rescue experiments. KIAA0586 encodes a novel protein, apparently specific to vertebrates, that localizes to the cytoplasm. We show that Gli3 processing is abnormal in talpid3 mutant cells but that Gli3 can still translocate to the nucleus. These results suggest that the talpid3 protein operates in the cytoplasm to regulate the activity of both Gli repressor and activator proteins. PMID- 16702411 TI - Very little intron loss/gain in Plasmodium: intron loss/gain mutation rates and intron number. AB - We compared intron positions in conserved regions of 3479 orthologous gene pairs from Plasmodium falciparum and Plasmodium yoelii, which likely diverged >or=100 million years ago (Mya). Only 27 out of 2212 positions were specific to one of the two species. Intron presence in related species shows that at least 19 and possibly 26 of the changes are due to intron loss, depending on phylogeny. The implied intron loss and gain rates are much lower than previously estimated for nematodes, arthropods, fungi, and plants, and are comparable only with the rates in vertebrates. That all observed changes were exact, occurring without loss or gain of flanking coding sequence, suggests intron loss via an mRNA intermediate, as does a nonsignificant trend toward loss of introns at adjacent positions. Many of the intron changes occurred in genes encoding proteins involved in nucleic acid-related processes, as previously found for intron gains in nematodes. Two changes occurred in the chloroquine resistance transporter, suggesting a role for positive selection in intron loss in Plasmodium. The dearth of intron loss and gain could be explained by the lack of known transposable elements in Plasmodium, since transposable elements and/or reverse transcriptase are thought to be necessary for both processes. The observed pattern suggests that the availability of stochastic intron loss and gain mutations can be a major determinant of changes in intron number. PMID- 16702412 TI - High-density haplotyping with microarray-based expression and single feature polymorphism markers in Arabidopsis. AB - Expression microarrays hybridized with RNA can simultaneously provide both phenotypic (gene expression) and genotypic (marker) data. We developed two types of genetic markers from Affymetrix GeneChip expression data to generate detailed haplotypes for 148 recombinant inbred lines (RILs) derived from Arabidopsis thaliana accessions Bayreuth and Shahdara. Gene expression markers (GEMs) are based on differences in transcript levels that exhibit bimodal distributions in segregating progeny, while single feature polymorphism (SFP) markers rely on differences in hybridization to individual oligonucleotide probes. Unlike SFPs, GEMs can be derived from any type of DNA-based expression microarray. Our method identifies SFPs independent of a gene's expression level. Alleles for each GEM and SFP marker were ascertained with GeneChip data from parental accessions as well as RILs; a novel algorithm for allele determination using RIL distributions capitalized on the high level of genetic replication per locus. GEMs and SFP markers provided robust markers in 187 and 968 genes, respectively, which allowed estimation of gene order consistent with that predicted from the Col-0 genomic sequence. Using microarrays on a population to simultaneously measure gene expression variation and obtain genotypic data for a linkage map will facilitate expression QTL analyses without the need for separate genotyping. We have demonstrated that gene expression measurements from microarrays can be leveraged to identify polymorphisms across the genome and can be efficiently developed into genetic markers that are verifiable in a large segregating RIL population. Both marker types also offer opportunities for massively parallel mapping in unsequenced and less studied species. PMID- 16702410 TI - Widespread genome duplications throughout the history of flowering plants. AB - Genomic comparisons provide evidence for ancient genome-wide duplications in a diverse array of animals and plants. We developed a birth-death model to identify evidence for genome duplication in EST data, and applied a mixture model to estimate the age distribution of paralogous pairs identified in EST sets for species representing the basal-most extant flowering plant lineages. We found evidence for episodes of ancient genome-wide duplications in the basal angiosperm lineages including Nuphar advena (yellow water lily: Nymphaeaceae) and the magnoliids Persea americana (avocado: Lauraceae), Liriodendron tulipifera (tulip poplar: Magnoliaceae), and Saruma henryi (Aristolochiaceae). In addition, we detected independent genome duplications in the basal eudicot Eschscholzia californica (California poppy: Papaveraceae) and the basal monocot Acorus americanus (Acoraceae), both of which were distinct from duplications documented for ancestral grass (Poaceae) and core eudicot lineages. Among gymnosperms, we found equivocal evidence for ancient polyploidy in Welwitschia mirabilis (Gnetales) and no evidence for polyploidy in pine, although gymnosperms generally have much larger genomes than the angiosperms investigated. Cross-species sequence divergence estimates suggest that synonymous substitution rates in the basal angiosperms are less than half those previously reported for core eudicots and members of Poaceae. These lower substitution rates permit inference of older duplication events. We hypothesize that evidence of an ancient duplication observed in the Nuphar data may represent a genome duplication in the common ancestor of all or most extant angiosperms, except Amborella. PMID- 16702413 TI - Cumulative mutations affecting sterol biosynthesis in the yeast Saccharomyces cerevisiae result in synthetic lethality that is suppressed by alterations in sphingolipid profiles. AB - UPC2 and ECM22 belong to a Zn(2)-Cys(6) family of fungal transcription factors and have been implicated in the regulation of sterol synthesis in Saccharomyces cerevisiae and Candida albicans. Previous reports suggest that double deletion of these genes in S. cerevisiae is lethal depending on the genetic background of the strain. In this investigation we demonstrate that lethality of upc2Delta ecm22Delta in the S288c genetic background is attributable to a mutation in the HAP1 transcription factor. In addition we demonstrate that strains containing upc2Delta ecm22Delta are also inviable when carrying deletions of ERG6 and ERG28 but not when carrying deletions of ERG3, ERG4, or ERG5. It has previously been demonstrated that UPC2 and ECM22 regulate S. cerevisiae ERG2 and ERG3 and that the erg2Delta upc2Delta ecm22Delta triple mutant is also synthetically lethal. We used transposon mutagenesis to isolate viable suppressors of hap1Delta, erg2Delta, erg6Delta, and erg28Delta in the upc2Delta ecm22Delta genetic background. Mutations in two genes (YND1 and GDA1) encoding apyrases were found to suppress the synthetic lethality of three of these triple mutants but not erg2Delta upc2Delta ecm22Delta. We show that deletion of YND1, like deletion of GDA1, alters the sphingolipid profiles, suggesting that changes in sphingolipids compensate for lethality produced by changes in sterol composition and abundance. PMID- 16702414 TI - Cis-transcriptional variation in maize inbred lines B73 and Mo17 leads to additive expression patterns in the F1 hybrid. AB - Microarray analysis of gene expression patterns in immature ear, seedling, and embryo tissues from the maize inbred lines B73 and Mo17 identified numerous genes with variable expression. Some genes had detectable expression in only one of the two inbreds; most of these genes were detected in the genomic DNA of both inbreds, indicating that the expression differences are likely caused by differential regulation rather than by differences in gene content. Gene expression was also monitored in the reciprocal F1 hybrids B73xMo17 and Mo17xB73. The reciprocal F1 hybrid lines did not display parental effects on gene expression levels. Approximately 80% of the differentially expressed genes displayed additive expression patterns in the hybrids relative to the inbred parents. The approximately 20% of genes that display nonadditive expression patterns tend to be expressed at levels within the parental range, with minimal evidence for novel expression levels greater than the high parent or less than the low parent. Analysis of allele-specific expression patterns in the hybrid suggested that intraspecific variation in gene expression levels is largely attributable to cis-regulatory variation in maize. Collectively, our data suggest that allelic cis-regulatory variation between B73 and Mo17 dictates maintenance of inbred allelic expression levels in the F1 hybrid, resulting in additive expression patterns. PMID- 16702415 TI - Exonuclease requirements for recombination of lambda-phage in recD mutants of Escherichia coli. AB - Recombination of lambda red gam phage in recD mutants is unaffected by inactivation of RecJ exonuclease. Since nucleases play redundant roles in E. coli, we inactivated several exonucleases in a recD mutant and discovered that 5' 3' exonuclease activity of RecJ and exonuclease VII is essential for lambda recombination, whereas exonucleases of 3'-5' polarity are dispensable. The implications of the presented data on current models for recombination initiation in E. coli are discussed. PMID- 16702416 TI - Toward a molecular understanding of pleiotropy. AB - Pleiotropy refers to the observation of a single gene influencing multiple phenotypic traits. Although pleiotropy is a common phenomenon with broad implications, its molecular basis is unclear. Using functional genomic data of the yeast Saccharomyces cerevisiae, here we show that, compared with genes of low pleiotropy, highly pleiotropic genes participate in more biological processes through distribution of the protein products in more cellular components and involvement in more protein-protein interactions. However, the two groups of genes do not differ in the number of molecular functions or the number of protein domains per gene. Thus, pleiotropy is generally caused by a single molecular function involved in multiple biological processes. We also provide genomewide evidence that the evolutionary conservation of genes and gene sequences positively correlates with the level of gene pleiotropy. PMID- 16702417 TI - The effect of gap length on double-strand break repair in Drosophila. AB - When a double-strand break has a gap between the broken ends, the missing information can be restored through synthesis from a homologous template. Here we address the question of how long such a gap can be before this process fails. We measured the frequency of homologous repair in the Drosophila germ line following the creation of gaps of specific sizes ranging from 3.8 to 210 kb. We found that gaps of >11 kb. PMID- 16702418 TI - Loss-of-function alleles of the JIL-1 kinase are strong suppressors of position effect variegation of the wm4 allele in Drosophila. AB - In this article we show that hypomorphic loss-of-function alleles of the JIL-1 histone H3S10 kinase are strong suppressors of position effect variegation (PEV) of the wm4 allele and that lack of JIL-1 activity can counteract the effect of the dominant enhancer Evar2-1 on PEV. PMID- 16702419 TI - Wild-derived XY sex-reversal mutants in the Medaka, Oryzias latipes. AB - The medaka, Oryzias latipes, has an XX/XY sex-determination mechanism. A Y-linked DM domain gene, DMY, has been isolated by positional cloning as a sex-determining gene in this species. Previously, we found 23 XY sex-reversed females from 11 localities by examining the genotypic sex of wild-caught medaka. Genetic analyses revealed that all these females had Y-linked gene mutations. Here, we aimed to clarify the cause of this sex reversal. To achieve this, we screened for mutations in the amino acid coding sequence of DMY and examined DMY expression at 0 days after hatching (dah) using densitometric semiquantitative RT-PCR. We found that the mutants could be classified into two groups. One contained mutations in the amino acid coding sequence of DMY, while the other had reduced DMY expression at 0 dah although the DMY coding sequence was normal. For the latter, histological analyses indicated that YwOurYwOur (YwOur, Y chromosome derived from an Oura XY female) individuals with the lowest DMY expression among the tested mutants were expected to develop into females at 0 dah. These results suggest that early testis development requires DMY expression above a threshold level. Mutants with reduced DMY expression may prove valuable for identifying DMY regulatory elements. PMID- 16702420 TI - Transcriptionally silenced transgenes in maize are activated by three mutations defective in paramutation. AB - Plants with mutations in one of three maize genes, mop1, rmr1, and rmr2, are defective in paramutation, an allele-specific interaction that leads to meiotically heritable chromatin changes. Experiments reported here demonstrate that these genes are required to maintain the transcriptional silencing of two different transgenes, suggesting that paramutation and transcriptional silencing of transgenes share mechanisms. We hypothesize that the transgenes are silenced through an RNA-directed chromatin mechanism, because mop1 encodes an RNA dependent RNA polymerase. In all the mutants, DNA methylation was reduced in the active transgenes relative to the silent transgenes at all of the CNG sites monitored within the transgene promoter. However, asymmetrical methylation persisted at one site within the reactivated transgene in the rmr1-1 mutant. With that one mutant, rmr1-1, the transgene was efficiently resilenced upon outcrossing to reintroduce the wild-type protein. In contrast, with the mop1-1 and rmr2-1 mutants, the transgene remained active in a subset of progeny even after the wild-type proteins were reintroduced by outcrossing. Interestingly, this immunity to silencing increased as the generations progressed, consistent with a heritable chromatin state being formed at the transgene in plants carrying the mop1-1 and rmr2-1 mutations that becomes more resistant to silencing in subsequent generations. PMID- 16702422 TI - The Hill-Robertson effect and the evolution of recombination. AB - In finite populations, genetic drift generates interference between selected loci, causing advantageous alleles to be found more often on different chromosomes than on the same chromosome, which reduces the rate of adaptation. This "Hill-Robertson effect" generates indirect selection to increase recombination rates. We present a new method to quantify the strength of this selection. Our model represents a new beneficial allele (A) entering a population as a single copy, while another beneficial allele (B) is sweeping at another locus. A third locus affects the recombination rate between selected loci. Using a branching process model, we calculate the probability distribution of the number of copies of A on the different genetic backgrounds, after it is established but while it is still rare. Then, we use a deterministic model to express the change in frequency of the recombination modifier, due to hitchhiking, as A goes to fixation. We show that this method can give good estimates of selection for recombination. Moreover, it shows that recombination is selected through two different effects: it increases the fixation probability of new alleles, and it accelerates selective sweeps. The relative importance of these two effects depends on the relative times of occurrence of the beneficial alleles. PMID- 16702423 TI - Mapping quantitative trait loci by an extension of the Haley-Knott regression method using estimating equations. AB - The Haley-Knott (HK) regression method continues to be a popular approximation to standard interval mapping (IM) of quantitative trait loci (QTL) in experimental crosses. The HK method is favored for its dramatic reduction in computation time compared to the IM method, something that is particularly important in simultaneous searches for multiple interacting QTL. While the HK method often approximates the IM method well in estimating QTL effects and in power to detect QTL, it may perform poorly if, for example, there is strong epistasis between QTL or if QTL are linked. Also, it is well known that the estimation of the residual variance by the HK method is biased. Here, we present an extension of the HK method that uses estimating equations based on both means and variances. For normally distributed phenotypes this estimating equation (EE) method is more efficient than the HK method. Furthermore, computer simulations show that the EE method performs well for very different genetic models and data set structures, including nonnormal phenotype distributions, nonrandom missing data patterns, varying degrees of epistasis, and varying degrees of linkage between QTL. The EE method retains key qualities of the HK method such as computational speed and robustness against nonnormal phenotype distributions, while approximating the IM method better in terms of accuracy and precision of parameter estimates and power to detect QTL. PMID- 16702421 TI - Developmental modulation of nonhomologous end joining in Caenorhabditis elegans. AB - Homologous recombination and nonhomologous end joining (NHEJ) are important DNA double-strand break repair pathways in many organisms. C. elegans strains harboring mutations in the cku-70, cku-80, or lig-4 NHEJ genes displayed multiple developmental abnormalities in response to radiation-induced DNA damage in noncycling somatic cells. These phenotypes did not result from S-phase, DNA damage, or mitotic checkpoints, apoptosis, or stress response pathways that regulate dauer formation. However, an additional defect in him-10, a kinetochore component, synergized with NHEJ mutations for the radiation-induced developmental phenotypes, suggesting that they may be triggered by mis-segregation of chromosome fragments. Although NHEJ was an important DNA repair pathway for noncycling somatic cells in C. elegans, homologous recombination was used to repair radiation-induced DNA damage in cycling somatic cells and in germ cells at all times. Noncycling germ cells that depended on homologous recombination underwent cell cycle arrest in G2, whereas noncycling somatic cells that depended on NHEJ arrested in G1, suggesting that cell cycle phase may modulate DNA repair during development. We conclude that error-prone NHEJ plays little or no role in DNA repair in C. elegans germ cells, possibly ensuring homology-based double strand break repair and transmission of a stable genome from one generation to the next. PMID- 16702424 TI - A novel approach for characterizing expression levels of genes duplicated by polyploidy. AB - Studying gene expression in polyploids is complicated by genomewide gene duplication and the problem of distinguishing transcript pools derived from each of the two homeologous genomes such as the A- and D-genomes of allotetraploid Gossypium. Short oligonucleotide probes designed to specifically target several hundred homeologous gene pairs of Gossypium were printed on custom NimbleGen microarrays. These results demonstrate that relative expression levels of homeologous genes may be measured by microarrays and that deviation from equal expression levels of homeologous loci may be common in the allotetraploid nucleus of Gossypium. PMID- 16702426 TI - Discovery and mapping of wheat Ph1 suppressors. AB - Pairing between wheat (Triticum turgidum and T. aestivum) homeologous chromosomes is prevented by the expression of the Ph1 locus on the long arm of chromosome 5B. The genome of Aegilops speltoides suppresses Ph1 expression in wheat x Ae. speltoides hybrids. Suppressors with major effects were mapped as Mendelian loci on the long arms of Ae. speltoides chromosomes 3S and 7S. The chromosome 3S locus was designated Su1-Ph1 and the chromosome 7S locus was designated Su2-Ph1. A QTL with a minor effect was mapped on the short arm of chromosome 5S and was designated QPh.ucd-5S. The expression of Su1-Ph1 and Su2-Ph1 increased homeologous chromosome pairing in T. aestivum x Ae. speltoides hybrids by 8.4 and 5.8 chiasmata/cell, respectively. Su1-Ph1 was completely epistatic to Su2-Ph1, and the two genes acting together increased homeologous chromosome pairing in T. aestivum x Ae. speltoides hybrids to the same level as Su1-Ph1 acting alone. QPh.ucd-5S expression increased homeologous chromosome pairing by 1.6 chiasmata/cell in T. aestivum x Ae. speltoides hybrids and was additive to the expression of Su2-Ph1. It is hypothesized that the products of Su1-Ph1 and Su2 Ph1 affect pairing between homeologous chromosomes by regulating the expression of Ph1 but the product of QPh.ucd-5S may primarily regulate recombination between homologous chromosomes. PMID- 16702425 TI - Natural history of transposition in the green alga Chlamydomonas reinhardtii: use of the AMT4 locus as an experimental system. AB - The AMT4 locus of the green alga Chlamydomonas reinhardtii, which we mapped to the long arm of chromosome 8, provides a good experimental system for the study of transposition. Most mutations that confer resistance to the toxic ammonium analog methylammonium are in AMT4 and a high proportion of spontaneous mutations are caused by transposon-related events. Among the 15 such events that we have characterized at the molecular level, 9 were associated with insertions of the retrotransposon TOC1, 2 with a small Gulliver-related transposon, and 1 with the Tcr1 transposon. We found that Tcr1 is apparently a foldback transposon with terminal inverted repeats that are much longer and more complex than previously realized. A duplication of Tcr1 yielded a configuration thought to be important for chromosomal evolution. Other mutations in AMT4 were caused by two mobile elements that have not been described before. The sequence of one, which we propose to call the Bill element, indicates that it probably transposes by way of a DNA intermediate and requires functions that it does not encode. The sequence of the other and bioinformatic analysis indicates that it derives from a miniature retrotransposon or TRIM, which we propose to call MRC1 (miniature retrotransposon of Chlamydomonas). PMID- 16702427 TI - Evolutionary divergence of exon flanks: a dissection of mutability and selection. AB - The intronic sequences flanking exon-intron junctions (i.e., exon flanks) are important for splice site recognition and pre-mRNA splicing. Recent studies show a higher degree of sequence conservation at flanks of alternative exons, compared to flanks of constitutive exons. In this article we performed a detailed analysis on the evolutionary divergence of exon flanks between human and chimpanzee, aiming to dissect the impact of mutability and selection on their evolution. Inside exon flanks, sites that might reside in ancestral CpG dinucleotides evolved significantly faster than sites outside of ancestral CpG dinucleotides. This result reflects a systematic variation of mutation rates (mutability) at exon flanks, depending on the local CpG contexts. Remarkably, we observed a significant reduction of the nucleotide substitution rate in flanks of alternatively spliced exons, independent of the site-by-site variation in mutability due to different CpG contexts. Our data provide concrete evidence for increased purifying selection at exon flanks associated with regulation of alternative splicing. PMID- 16702428 TI - PAPPA2, an enzyme that cleaves an insulin-like growth-factor-binding protein, is a candidate gene for a quantitative trait locus affecting body size in mice. AB - Identifying genes responsible for quantitative variation remains a major challenge. We previously identified a quantitative trait locus (QTL) affecting body size that segregated between two inbred strains of mice. By fine mapping, we have refined the location of this QTL to a genomic region containing only four protein-coding genes. One of these genes, PAPPA2, is a strong candidate because it codes for an enzyme that cleaves insulin-like growth-factor-binding protein 5 (IGFBP-5), an important stimulator of bone formation. Among littermates that segregate only for the four-gene region, we show that the QTL has a significant effect on the circulating levels of IGFBP-5 and IGFBP-3 (the latter subject to limited degradation by PAPPA2), but not on levels of IGFBP-2 and IGFBP-4, which are not cleaved by PAPPA2. There are 14 nonsynonymous SNPs among QTL alleles, which may affect the activity of the translated protein. The refinement of the target region to four genes and the finding that the QTL affects IGFBP-5 levels suggest that PAPPA2 may be involved with normal postnatal growth. Our mapping results also illustrate the potentially fractal nature of QTL: as we mapped our QTL with increasing resolution, what appeared to be a single QTL resolved into three closely linked QTL (previous work), and then one of these was further dissected into two in this study. PMID- 16702429 TI - A clarification of the Hardy-Weinberg law. AB - C. C. Li showed that Hardy-Weinberg proportions (HWP) can be maintained in a large population by nonrandom mating as well as random mating. In particular he gave the mating matrix for the symmetric case in the most general form possible. Thus Li showed that, once HWP are attained, the same proportions can be maintained by what he called pseudorandom mating. This article shows that, starting from any genotypic distribution at a single locus with two alleles, the same in each sex, HWP can be reached in one round of nonrandom mating with no change in allele frequency. In the model that demonstrates this fact, random mating is represented by a single point in a continuum of nonrandom possibilities. PMID- 16702431 TI - Natural variation in MAM within and between populations of Arabidopsis lyrata determines glucosinolate phenotype. AB - The genetic variation that underlies the glucosinolate phenotype of Arabidopsis lyrata ssp. petraea was investigated between and within populations. A candidate glucosinolate biosynthetic locus (MAM, containing methylthioalkylmalate synthase genes) was mapped in A. lyrata to a location on linkage group 6 corresponding to the homologous location for MAM in A. thaliana. In A. thaliana MAM is responsible for side chain elongation in aliphatic glucosinolates, and the MAM phenotype can be characterized by the ratios of long- to short-chain glucosinolates. A quantitative trait loci (QTL) analysis of glucosinolate ratios in an A. lyrata interpopulation cross found one QTL at MAM. Additional QTL were identified for total indolic glucosinolates and for the ratio of aliphatic to indolic glucosinolates. MAM was then used as the candidate gene for a within-population cosegregation analysis in a natural A. lyrata population from Germany. Extensive variation in microsatellite markers at MAM was found and this variation cosegregated with the same glucosinolate ratios as in the QTL study. The combined results indicate that both between- and within-population genetic variation in the MAM region determines phenotypic variation in glucosinolate side chains in A. lyrata. PMID- 16702430 TI - Rapid evolution of major histocompatibility complex class I genes in primates generates new disease alleles in humans via hitchhiking diversity. AB - A plausible explanation for many MHC-linked diseases is lacking. Sequencing of the MHC class I region (coding units or full contigs) in several human and nonhuman primate haplotypes allowed an analysis of single nucleotide variations (SNV) across this entire segment. This diversity was not evenly distributed. It was rather concentrated within two gene-rich clusters. These were each centered, but importantly not limited to, the antigen-presenting HLA-A and HLA-B/-C loci. Rapid evolution of MHC-I alleles, as evidenced by an unusually high number of haplotype-specific (hs) and hypervariable (hv) (which could not be traced to a single species or haplotype) SNVs within the classical MHC-I, seems to have not only hitchhiked alleles within nearby genes, but also hitchhiked deleterious mutations in these same unrelated loci. The overrepresentation of a fraction of these hvSNV (hv1SNV) along with hsSNV, as compared to those that appear to have been maintained throughout primate evolution (trans-species diversity; tsSNV; included within hv2SNV) tends to establish that the majority of the MHC polymorphism is de novo (species specific). This is most likely reminiscent of the fact that these hsSNV and hv1SNV have been selected in adaptation to the constantly evolving microbial antigenic repertoire. PMID- 16702432 TI - Analysis of the proteins involved in the in vivo repair of base-base mismatches and four-base loops formed during meiotic recombination in the yeast Saccharomyces cerevisiae. AB - DNA mismatches are generated when heteroduplexes formed during recombination involve DNA strands that are not completely complementary. We used tetrad analysis in Saccharomyces cerevisiae to examine the meiotic repair of a base-base mismatch and a four-base loop in a wild-type strain and in strains with mutations in genes implicated in DNA mismatch repair. Efficient repair of the base-base mismatch required Msh2p, Msh6p, Mlh1p, and Pms1p, but not Msh3p, Msh4p, Msh5p, Mlh2p, Mlh3p, Exo1p, Rad1p, Rad27p, or the DNA proofreading exonuclease of DNA polymerase delta. Efficient repair of the four-base loop required Msh2p, Msh3p, Mlh1p, and Pms1p, but not Msh4p, Msh5p, Msh6p, Mlh2p, Mlh3p, Exo1p, Rad1p, Rad27p, or the proofreading exonuclease of DNA polymerase delta. We find evidence that a novel Mlh1p-independent complex competes with an Mlhp-dependent complex for the repair of a four-base loop; repair of the four-base loop was affected by loss of the Mlh3p, and the repair defect of the mlh1 and pms1 strains was significantly smaller than that observed in the msh2 strain. We also found that the frequency and position of local double-strand DNA breaks affect the ratio of mismatch repair events that lead to gene conversion vs. restoration of Mendelian segregation. PMID- 16702434 TI - Parent-of-origin effects on mRNA expression in Drosophila melanogaster not caused by genomic imprinting. AB - Parent-of-origin effects create differences in gene expression among genetically identical individuals. Using measurements of allele-specific expression, we demonstrate that previously reported parent-of-origin effects on standing mRNA levels in Drosophila melanogaster are not attributable to genomic imprinting. Offspring from reciprocal crosses exhibit differences in total expression without differences in allelic expression, indicating that other types of maternal and/or paternal effects alter expression. PMID- 16702433 TI - High-resolution mapping of quantitative trait loci affecting increased life span in Drosophila melanogaster. AB - Limited life span and senescence are near-universal characteristics of eukaryotic organisms, controlled by many interacting quantitative trait loci (QTL) with individually small effects, whose expression is sensitive to the environment. Analyses of mutations in model organisms have shown that genes affecting stress resistance and metabolism affect life span across diverse taxa. However, there is considerable segregating variation for life span in nature, and relatively little is known about the genetic basis of this variation. Replicated lines of Drosophila that have evolved increased longevity as a correlated response to selection for postponed senescence are valuable resources for identifying QTL affecting naturally occurring variation in life span. Here, we used deficiency complementation mapping to identify at least 11 QTL on chromosome 3 that affect variation in life span between five old (O) lines selected for postponed senescence and their five base (B) population control lines. Most QTL were sex specific, and all but one affected multiple O lines. The latter observation is consistent with alleles at intermediate frequency in the base population contributing to the response to selection for postponed senescence. The QTL were mapped with high resolution and contained from 12 to 170 positional candidate genes. PMID- 16702435 TI - Functional implication of an Arg307Gly substitution in corticosteroid-binding globulin, a candidate gene for a quantitative trait locus associated with cortisol variability and obesity in pig. AB - We previously reported that corticosteroid-binding globulin gene (Cbg) may be the causal gene of a quantitative trait locus associated with cortisol levels, fat deposition, and muscle content in a pig intercross. Sequence analysis of parental animals allowed us to identify four amino-acid substitutions. Here we have examined if any of these single amino acid substitutions could be responsible for the difference in CBG binding and affinity for cortisol between the parental breeds, using in vitro assays of Cbg variants after transfection of mammalian cells. Additionally, the Cbg coding region was analyzed in samples from a synthetic pig line to study association between polymorphism and CBG biochemical properties, carcass composition, and meat quality. Both in vitro transfection assays and the association studies suggest a role of the Arg307Gly mutation in increasing CBG capacity (by >70%) and decreasing CBG affinity for cortisol (by 30%). The Ile265Val substitution may also have an effect on decreasing CBG affinity for cortisol by 25%. The mutations Ser15Ile and Thr257Met do not seem to have an effect on CBG parameters. The Arg307Gly substitution was the only mutation associated with a parameter of meat quality and no mutation was linked to carcass composition. PMID- 16702436 TI - Signatures of demographic history and natural selection in the human major histocompatibility complex Loci. AB - Many lines of evidence show that several HLA loci have experienced balancing selection. However, distinguishing among demographic and selective explanations for patterns of variation observed with HLA genes remains a challenge. In this study we address this issue using data from a diverse set of human populations at six classical HLA loci and, employing a comparative genomics approach, contrast results for HLA loci to those for non-HLA markers. Using a variety of analytic methods, we confirm and extend evidence for selection acting on several HLA loci. We find that allele frequency distributions for four of the six HLA loci deviate from neutral expectations and show that this is unlikely to be explained solely by demographic factors. Other features of HLA variation are explained in part by demographic history, including decreased heterozygosity and increased LD for populations at greater distances from Africa and a similar apportionment of genetic variation for HLA loci compared to putatively neutral non-HLA loci. On the basis of contrasts among different HLA loci and between HLA and non-HLA loci, we conclude that HLA loci bear detectable signatures of both natural selection and demographic history. PMID- 16702437 TI - Fine-scale map of encyclopedia of DNA elements regions in the Korean population. AB - The International HapMap Project aims to generate detailed human genome variation maps by densely genotyping single-nucleotide polymorphisms (SNPs) in CEPH, Chinese, Japanese, and Yoruba samples. This will undoubtedly become an important facility for genetic studies of diseases and complex traits in the four populations. To address how the genetic information contained in such variation maps is transferable to other populations, the Korean government, industries, and academics have launched the Korean HapMap project to genotype high-density Encyclopedia of DNA Elements (ENCODE) regions in 90 Korean individuals. Here we show that the LD pattern, block structure, haplotype diversity, and recombination rate are highly concordant between Korean and the two HapMap Asian samples, particularly Japanese. The availability of information from both Chinese and Japanese samples helps to predict more accurately the possible performance of HapMap markers in Korean disease-gene studies. Tagging SNPs selected from the two HapMap Asian maps, especially the Japanese map, were shown to be very effective for Korean samples. These results demonstrate that the HapMap variation maps are robust in related populations and will serve as an important resource for the studies of the Korean population in particular. PMID- 16702438 TI - Parallel changes in global protein profiles during long-term experimental evolution in Escherichia coli. AB - Twelve populations of Escherichia coli evolved in and adapted to a glucose limited environment from a common ancestor. We used two-dimensional protein electrophoresis to compare two evolved clones, isolated from independently derived populations after 20,000 generations. Exceptional parallelism was detected. We compared the observed changes in protein expression profiles with previously characterized global transcription profiles of the same clones; this is the first time such a comparison has been made in an evolutionary context where these changes are often quite subtle. The two methodologies exhibited some remarkable similarities that highlighted two different levels of parallel regulatory changes that were beneficial during the evolution experiment. First, at the higher level, both methods revealed extensive parallel changes in the same global regulatory network, reflecting the involvement of beneficial mutations in genes that control the ppGpp regulon. Second, both methods detected expression changes of identical gene sets that reflected parallel changes at a lower level of gene regulation. The protein profiles led to the discovery of beneficial mutations affecting the malT gene, with strong genetic parallelism across independently evolved populations. Functional and evolutionary analyses of these mutations revealed parallel phenotypic decreases in the maltose regulon expression and a high level of polymorphism at this locus in the evolved populations. PMID- 16702439 TI - Inferring population parameters from single-feature polymorphism data. AB - This article is concerned with a statistical modeling procedure to call single feature polymorphisms from microarray experiments. We use this new type of polymorphism data to estimate the mutation and recombination parameters in a population. The mutation parameter can be estimated via the number of single feature polymorphisms called in the sample. For the recombination parameter, a two-feature sampling distribution is derived in a way analogous to that for the two-locus sampling distribution with SNP data. The approximate-likelihood approach using the two-feature sampling distribution is examined and found to work well. A coalescent simulation study is used to investigate the accuracy and robustness of our method. Our approach allows the utilization of single-feature polymorphism data for inference in population genetics. PMID- 16702440 TI - Pharmacological characterization of hydrolysis-resistant analogs of oleoylethanolamide with potent anorexiant properties. AB - Oleoylethanolamide (OEA) is an endogenous lipid mediator that reduces food intake, promotes lipolysis, and decreases body weight gain in rodents by activating peroxisome proliferator-activated receptor-alpha (PPAR-alpha). The biological effects of OEA are terminated by two intracellular lipid hydrolase enzymes, fatty-acid amide hydrolase and N-acylethanolamine-hydrolyzing acid amidase. In the present study, we describe OEA analogs that resist enzymatic hydrolysis, activate PPAR-alpha with high potency in vitro, and persistently reduce feeding when administered in vivo either parenterally or orally. The most potent of these compounds, (Z)-(R)-9-octadecenamide,N-(2-hydroxyethyl,1-methyl) (KDS-5104), stimulates transcriptional activity of PPAR-alpha with a half-maximal effective concentration (EC50) of 100 +/- 21 nM (n = 11). Parenteral administration of KDS-5104 in rats produces persistent dose-dependent prolongation of feeding latency and postmeal interval (half-maximal effective dose, ED50 = 2.4 +/- 1.8 mg kg(-1) i.p.; n = 18), as well as increased and protracted tissue exposure compared with OEA. Oral administration of the compound also results in a significant tissue exposure and reduction of food intake in free-feeding rats. These results suggest that the endogenous high-affinity PPAR alpha agonist OEA may provide a scaffold for the discovery of novel orally active PPAR-alpha ligands. PMID- 16702441 TI - Interaction of methotrexate with organic-anion transporting polypeptide 1A2 and its genetic variants. AB - Methotrexate (MTX) is used in patients with malignant and autoimmune diseases. This drug is primarily excreted unchanged in the urine, and its net excretion occurs via active secretory and reabsorptive processes. We characterized the interaction of MTX with human organic-anion transporting polypeptide transporter (OATP) 1A2, which is expressed in tissues important for MTX disposition and toxicity, such as the intestine, kidney, liver, and endothelial cells of the blood-brain barrier. In Xenopus laevis oocytes expressing OATP1A2, the uptake of the model substrate, estrone-3-sulfate (ES), was enhanced 30-fold compared with uninjected oocytes. MTX uptake in oocytes expressing OATP1A2 was saturable (Km = 457 +/- 118 microM; Vmax = 17.5 +/- 4.9 pmol/oocyte/60 min) and sensitive to extracellular pH. That is, acidic pHs stimulated MTX uptake by as much as 7-fold. Seven novel protein-altering variants were identified in 270 ethnically diverse DNA samples. Four protein-altering variants in OATP1A2 exhibited altered transport of ES and/or MTX. The common variant, protein reference sequence (p.) Ile13Thr, was hyperfunctional for ES and MTX and showed a 2-fold increase in the V(max) for ES. The common variant, p. Glu172Asp, exhibited reduced maximal transport capacity for ES and MTX. p. Arg168Cys was hypofunctional, and p. Asn277DEL was nonfunctional. Because of its expression on the apical membrane of the distal tubule and in tissues relevant to MTX disposition and toxicity, these findings suggest that OATP1A2 may play a role in active tubular reabsorption of MTX and in MTX-induced toxicities. Furthermore, genetic variation in OATP1A2 may contribute to variation in MTX disposition and response. PMID- 16702442 TI - Time-dependent effects of haloperidol and ziprasidone on nerve growth factor, cholinergic neurons, and spatial learning in rats. AB - In this rodent study, we evaluated the effects of different time periods (7, 14, 45, and 90 days) of oral treatment with haloperidol (HAL; 2.0 mg/kg/day) or ziprasidone (ZIP; 12.0 mg/kg/day) on nerve growth factor (NGF) and choline acetyltransferase (ChAT) levels in the hippocampus, and we subsequently assessed water maze task performance, prepulse inhibition (PPI) of the auditory gating response, and several NGF-related proteins and cholinergic markers after 90 days of treatment. Seven and 14 days of treatment with either HAL or ZIP resulted in a notable increase in NGF and ChAT immunoreactivity in the dentate gyrus (DG), CA1, and CA3 areas of the hippocampus. After 45 days, NGF and ChAT immunoreactivity had abated to control levels in ZIP-treated animals, but it was markedly reduced in HAL-treated subjects. After 90 days of treatment, NGF and ChAT levels were substantially lower than controls in both antipsychotic groups. Furthermore, after 90 days of treatment and a drug-free washout period, water maze performance (but not PPI) was impaired in both antipsychotic groups, although the decrement was greater in the HAL group. Several NGF-related and cholinergic proteins were diminished in the brains of subjects treated with either neuroleptic as well. These data support the premise that, although ZIP (given chronically) seems somewhat superior to HAL due to less pronounced behavioral effects and a more delayed appearance of neurochemical deficits, both antipsychotics produce time dependent deleterious effects on NGF, cholinergic markers (i.e., important neurobiological substrates of memory), and cognitive function. PMID- 16702443 TI - Discovery and characterization of triaminotriazine aniline amides as highly selective p38 kinase inhibitors. AB - The p38 mitogen-activated protein (MAP) kinases are a family of serine/threonine protein kinases that play important roles in cellular responses to inflammation and external stress. Inhibitors of the p38 MAP kinase have shown promise for potential treatment of inflammatory disorders such as rheumatoid arthritis, acute coronary syndrome, psoriasis, and Crohn's disease. We identified a novel class of p38 inhibitors via high-throughput screening. PS200981 [3-(4-(1,4-diazepan-1-yl) 6-(((1S,2R,5S)-6,6-dimethylbicyclo[3.1.1]heptan-2-yl)methylamino)-1,3,5-triazin-2 ylamino)-4-methylbenzamide], a representative compound identified from screening a collection of combinatorial libraries, amounting to 2.1 million compounds, inhibits p38alpha kinase and the lipopolysaccharide (LPS)-induced increase in tumor necrosis factor (TNF) alpha levels in cell media of human monocytes with IC50 values of 1 microM. The screening data revealed a preferred synthon, 3-amino 4-methyl benzamide, which is critical for the activity against p38. This synthon appeared almost exclusively in screening hits including PS200981, and slight variations of this synthon including 3-amino benzamide and 2-amino-4-methyl benzamide also contained in the library were inactive. PS200981 is equally potent against the alpha and beta forms of p38 but did not inhibit p38 gamma and is >25 fold selective versus a panel of other kinases. PS200981 inhibited the LPS induced increase in TNFalpha levels when administered at 30 mg/kg to mice. Selectivity and in vivo activity of this class of p38 inhibitors was further demonstrated by PS166276 [(R)-3-(4-(isobutyl(methyl)-amino)-6-(pyrrolidin-3 ylamino)-1,3,5-triazin-2-ylamino)-4-methylbenzamide], a highly structurally related but more potent and less cytotoxic inhibitor, in several intracellular signaling assays, and in LPS-challenged mice. Overall, this novel class of p38 inhibitors is potent, active in vitro and in vivo, and is highly selective. PMID- 16702444 TI - CT enterography as a diagnostic tool in evaluating small bowel disorders: review of clinical experience with over 700 cases. AB - Computed tomographic (CT) enterography combines the improved spatial and temporal resolution of multi-detector row CT with large volumes of ingested neutral enteric contrast material to permit visualization of the small bowel wall and lumen. Adequate luminal distention can usually be achieved with oral hyperhydration, thereby obviating nasoenteric intubation and making CT enterography a useful, well-tolerated study for the evaluation of diseases affecting the mucosa and bowel wall. Unlike routine CT, which has been used to detect the extraenteric complications of Crohn disease such as fistula and abscess, CT enterography clearly depicts the small bowel inflammation associated with Crohn disease by displaying mural hyperenhancement, stratification, and thickening; engorged vasa recta; and perienteric inflammatory changes. As a result, CT enterography is becoming the first-line modality for the evaluation of suspected inflammatory bowel disease. CT enterography has also become an important alternative to traditional fluoroscopy in the assessment of other small bowel disorders such as celiac sprue and small bowel neoplasms. PMID- 16702446 TI - Dynamic secretin-enhanced MR cholangiopancreatography. AB - Secretin causes temporary dilatation of pancreatic ducts, principally by increasing pancreatic exocrine secretions, and thus allows better visualization of the ducts at magnetic resonance (MR) cholangiopancreatography. Secretin enhanced MR cholangiopancreatography is useful for detection and diagnosis of a variety of congenital, inflammatory, and neoplastic conditions of the pancreas. Although MR cholangiopancreatography without secretin is a reliable method for evaluating the pancreatobiliary ductal system, the authors believe that secretin enhanced MR cholangiopancreatography gives additional valuable functional and anatomic information about the pancreatic duct and pancreatic excretory capacity. PMID- 16702447 TI - Review of the abdominal manifestations of cystic fibrosis in the adult patient. AB - Cystic fibrosis is a common inherited fatal disease. As the life expectancy of affected individuals continues to increase with advances in disease management, this disease is no longer limited to the pediatric population. Currently, 40% of patients with cystic fibrosis are adults. In addition, patients may not present until adulthood and frequently have extrapulmonary symptoms. Abdominal manifestations are common and affect multiple organ systems. Hepatobiliary manifestations include fatty infiltration of the liver, gallbladder abnormalities, bile duct abnormalities, focal biliary fibrosis, and multinodular cirrhosis. Manifestations in the pancreas include acute pancreatitis, fatty replacement, calcifications, cysts, duct abnormalities, and carcinoma. Gastrointestinal manifestations include gastroesophageal reflux, peptic ulceration of the gastric and duodenal mucosa, distal intestinal obstruction syndrome, intussusception, appendicitis, fibrosing colonopathy, pneumatosis intestinalis, rectal mucosal prolapse, malignancies, and pseudomembranous colitis. Renal manifestations include nephrolithiasis, as well as secondary renal complications such as interstitial nephritis due to antibiotic therapy and amyloidosis. Awareness of these manifestations is important to successfully guide management of cystic fibrosis in adult patients. PMID- 16702448 TI - Anterior abdominal wall hernias: findings in barium studies. AB - Findings of anterior abdominal wall hernias at computed tomography and magnetic resonance imaging are well documented; however, little information is available about the depiction and characterization of such hernias in barium studies, primarily in small-bowel follow-through examinations. Such examinations are performed frequently, and radiologists should be familiar with the hernia features that may be observed. Anterior abdominal wall hernias are best recognized in profile on lateral spot images from a small-bowel follow-through study when one or more loops of bowel extend beyond the fascial planes of the anterior abdominal wall, with luminal narrowing at the entry or exit site of the hernia or at both sites. In some patients, the hernia also can be recognized indirectly on a frontal view because of the displacement and, often, extrinsic compression or deformity of herniated bowel loops. In such cases, additional views should be obtained with the patient in the lateral position to confirm the presence of the hernia with direct visualization of the herniated loops in profile. The reducibility of bowel from an anterior abdominal wall hernia also can be assessed with manual palpation of the abdominal wall while the patient is in the lateral position. Manual palpation performed during fluoroscopy helps determine whether the bowel loops can be returned to the proper location or are fixed in the hernia, an important observation because of the higher risk of obstruction or strangulation when the bowel is incarcerated. Fluoroscopy therefore is a useful technique for the detection and characterization of anterior abdominal wall hernias in barium studies. PMID- 16702449 TI - Preoperative staging of rectal cancer with MR Imaging: correlation with surgical and histopathologic findings. AB - Rectal cancer is a common malignancy that continues to have a highly variable outcome, with local pelvic recurrence after surgical resection usually leading to incurable disease. The success of tumor excision depends largely upon accurate tumor staging and appropriate surgical technique, although the results of recent surgical trials indicate that evaluation of the involvement of the mesorectal fat and mesorectal fascia is even more important than T staging for treatment planning. Magnetic resonance (MR) imaging is increasingly being used to evaluate tumor resectability in patients with rectal cancer and to determine which patients can be treated with surgery alone and which will require radiation therapy to promote tumor regression. High-spatial-resolution MR imaging has proved useful in clarifying the relationship between a tumor and the mesorectal fascia, which represents the circumferential resection margin at total mesorectal excision. Phased-array surface coil MR imaging in particular plays a vital role in the therapeutic management of rectal cancer. At present, phased-array MR imaging best fulfills the clinical requirements for preoperative staging of rectal cancer. However, preoperative evaluation of the other prognostic factor, nodal status, is still problematic, and further studies will be needed to better define the role of MR imaging in this context. PMID- 16702450 TI - Multimodality imaging of pancreatic and biliary congenital anomalies. AB - A wide spectrum of anomalies of the pancreas, the pancreatic ductal system, and the biliary tree are commonly encountered at radiologic evaluation. These anomalies may simulate various neoplastic, inflammatory, and posttraumatic conditions and should be part of the differential diagnosis for a variety of abnormalities found at diagnostic imaging. Anatomic variants, developmental anomalies (eg, pancreas divisum, annular pancreas, ectopic pancreas, pancreatic agenesis and hypoplasia), and congenital diseases (congenital pancreatic cysts, von Hippel-Lindau disease, choledochal cysts), in addition to potential imaging pitfalls (uneven distribution of fat, "pseudomasses"), can all pose a diagnostic challenge for the radiologist. Familiarity with these anomalies, the imaging techniques available for their study, and their variable imaging manifestations is necessary for differentiating them from other biliary and pancreatic conditions. A basic understanding of the embryologic development and normal anatomy of the pancreas and biliary tree is also essential for identifying these anomalies. PMID- 16702451 TI - Adult intestinal intussusception: CT appearances and identification of a causative lead point. AB - The widespread application of computed tomography (CT) in different clinical situations has increased the detection of intussusception, particularly non-lead point intussusception, which tends to be transient. Consequently, determining the clinical significance of intussusception seen at CT poses a diagnostic challenge. Patients with intussusception may or may not be symptomatic, and symptoms can be acute, intermittent, or chronic, making clinical diagnosis difficult. In most cases, radiologists can readily make the correct diagnosis of intestinal intussusception by noting the typical bowel-within-bowel appearance at abdominal CT. However, the CT findings that help differentiate between lead point and non lead point intussusception have not been well studied. Nevertheless, although there is considerable overlap of CT findings, when a lead mass is seen at CT as a separate and distinct entity vis-a-vis edematous bowel, it can be considered a reliable indicator of a lead point intussusception. Differentiating between lead point and non-lead point intussusception is important in determining the appropriate treatment and has the potential to reduce the prevalence of unnecessary surgery. PMID- 16702452 TI - US of the major salivary glands: anatomy and spatial relationships, pathologic conditions, and pitfalls. AB - Ultrasonography (US) is useful for differential diagnosis of diseases of the salivary glands. In acute inflammation, salivary glands are enlarged and hypoechoic with increased blood flow; they may contain multiple small, oval, hypoechoic areas. In chronic inflammation, salivary glands are normal sized or smaller, hypoechoic, and inhomogeneous. Sialolithiasis appears as markedly hyperechoic lines or points with distal acoustic shadowing. Sialosis appears as enlarged hyperechoic glands without focal lesions or increased blood flow. The US features of advanced Sjogren syndrome include inhomogeneous salivary glands with scattered small, oval, hypoechoic or anechoic areas, usually well defined, and increased parenchymal blood flow. Pleomorphic adenomas are usually hypoechoic, well-defined, lobulated lesions with posterior acoustic enhancement that may contain calcifications; Warthin tumors are usually oval, hypoechoic, well-defined lesions that often contain anechoic areas and are often hypervascularized. Malignant neoplasms of the salivary glands may have irregular shapes, irregular borders, blurred margins, and a hypoechoic inhomogeneous structure or may have a benign appearance. Salivary gland cysts have well-defined margins, anechoic contents, posterior acoustic enhancement, and no internal blood flow. However, US appearances of some diseases may overlap, thus producing diagnostic pitfalls. PMID- 16702453 TI - MR imaging of temporomandibular joint dysfunction: a pictorial review. AB - Temporomandibular joint (TMJ) dysfunction is a common condition that is best evaluated with magnetic resonance (MR) imaging. The first step in MR imaging of the TMJ is to evaluate the articular disk, or meniscus, in terms of its morphologic features and its location relative to the condyle in both closed- and open-mouth positions. Disk location is of prime importance because the presence of a displaced disk is a critical sign of TMJ dysfunction. However, disk displacement is also frequently seen in asymptomatic volunteers, so that other findings may be required to help make the diagnosis. These findings include thickening of an attachment of the lateral pterygoid muscle, rupture of retrodiskal layers, and joint effusion and can serve as indirect early signs of TMJ dysfunction. It is important for the radiologist to detect early MR imaging signs of dysfunction, thereby avoiding the evolution of this condition to its final stage, an advanced and irreversible phase that is characterized by osteoarthritic changes such as condylar flattening or osteophytes. Further studies conducted with the latest MR imaging techniques will allow a better understanding of the sources of TMJ pain and of any discrepancy between imaging findings and patient symptoms. PMID- 16702454 TI - Diagnosis of midface fractures with CT: what the surgeon needs to know. AB - Computed tomography is commonly used to evaluate patients with blunt facial trauma. With the high definition of the current scanners, even small fractures of the facial skeleton can be visualized. In complex midface injuries, it can be difficult for the radiologist to know which fractures are important to point out to the surgeon. An understanding of the anatomically relevant and surgically accessible craniofacial buttresses is critical for management of these injuries. Naso-orbitoethmoid fractures are classified according to the degree of injury to the medial canthal attachment. If the nasofrontal ducts are disrupted, surgical obliteration of the frontal sinus is needed to prevent formation of a mucocele. Displaced fractures of the zygomaticomaxillary complex often increase orbital volume due to angulation of the lateral orbital wall at the zygomaticosphenoid suture. If the zygomatic arch is severely comminuted or angulated, surgical exposure is indicated. In orbital fractures, the position and shape of the medial and inferior rectus muscles can indicate whether entrapment and diplopia are likely. Pediatric "trapdoor" orbital fractures and fractures of the orbital apex associated with decreasing vision represent surgical emergencies. Le Fort fractures involve disruption of the pterygoid plates from the posterior maxilla; any combination of Le Fort I, II, and III patterns can occur. PMID- 16702455 TI - Delayed enhancement MR imaging: utility in myocardial assessment. AB - Use of magnetic resonance (MR) imaging for diagnosis of cardiac diseases and treatment monitoring is expanding. Delayed myocardial enhancement MR imaging is performed after administration of paramagnetic contrast agents and is used for a growing number of clinical applications. This technique was developed primarily for characterization of myocardial scarring after myocardial infarction. On delayed enhancement MR images, scarring or fibrosis appears as an area of high signal intensity that is typically subendocardial or transmural in a coronary artery distribution. However, delayed myocardial enhancement is not specific for myocardial infarction and can occur in a variety of other disorders, such as inflammatory or infectious diseases of the myocardium, cardiomyopathy, cardiac neoplasms, and congenital or genetic cardiac conditions, as well as after cardiac interventions. In nonischemic myocardial disease, the delayed enhancement usually does not occur in a coronary artery distribution and is often midwall rather than subendocardial or transmural. Therefore, the patient's clinical history is critical in the evaluation of delayed myocardial enhancement MR images. PMID- 16702458 TI - Pediatric presacral masses. AB - Various types of masses may affect the presacral area in children. A presacral mass may be congenital or developmental or may arise from inflammation. The mass may have neural, vascular, lymphatic, or mesenchymal origins and may be primary (as in focal disease) or systemic (as in multifocal disease). Because the clinical manifestations of presacral masses are often nonspecific, imaging plays an important role in the detection and differentiation of these masses. Information obtained from imaging is also critical for management, especially for surgical planning. For these reasons, it is important that radiologists be familiar with the anatomy of the presacral region and with the imaging features of the various lesions that may occur in this region in children. For the accurate interpretation of findings, radiologists also must know the specific advantages and limitations of each of the imaging modalities used to evaluate this category of abnormalities. PMID- 16702456 TI - Obstetric US: watch the fetal hands. AB - Hand anomalies are difficult to diagnose and are often overlooked during prenatal ultrasonography (US). The spectrum of malformations varies from subtle finger deformities to the complete amputation of limbs. Malformations of the hand can be classified, according to the predominant anomaly, among the following categories: alignment abnormalities (clenched hand, camptodactyly, clinodactyly, hypokinesia, clubhand, phocomelia), thumb anomalies, abnormal size (macrodactyly, trident hand), abnormal echogenicity (abnormal calcifications), abnormal number (polydactyly, syndactyly, ectrodactyly), and constriction band sequence. A fetal hand anomaly has important diagnostic and prognostic implications as well as functional consequences. Malformation may be isolated but often is associated with a syndrome or karyotype anomaly. Classification and characterization of the anomaly help to narrow the differential diagnosis: Some malformations (clenched hand, hitchhiker thumb) are highly suggestive of a specific diagnosis. The detection of a fetal hand malformation warrants a complete work-up, including complete fetal and cardiac US examinations, as well as genetic counseling to determine whether familial inquiry and karyotype analysis are necessary. PMID- 16702459 TI - Combined labeled leukocyte and technetium 99m sulfur colloid bone marrow imaging for diagnosing musculoskeletal infection. AB - The use of labeled leukocyte (white blood cell [WBC]) studies in the diagnosis of osteomyelitis can be problematic. A combined study consisting of WBC imaging and complementary bone marrow imaging performed with technetium 99m (99mTc) sulfur colloid is approximately 90% accurate and is especially useful for diagnosing osteomyelitis in situations involving altered marrow distribution. There are limitations and pitfalls associated with a combined study. If there is no labeled WBC activity in the region of interest, marrow imaging is not useful. The sulfur colloid image becomes photopenic within about 1 week after the onset of infection, so that the study should be interpreted cautiously in the acute setting. Labeled WBC accumulation in lymph nodes can also confound image interpretation, although nodal activity can usually be recognized because it is typically round, discrete, multifocal, linear in distribution, and often bilateral. Furthermore, 99mTc-sulfur colloid that is improperly prepared or is more than about 2 hours old degrades image quality, potentially causing erroneous conclusions. Nevertheless, WBC-marrow imaging is a very accurate technique for diagnosing osteomyelitis. Knowledge of the criteria for image interpretation and of the aforementioned limitations and pitfalls, combined with careful attention to imaging technique, will maximize the value of this study. PMID- 16702460 TI - Tumoral calcinosis: pearls, polemics, and alternative possibilities. AB - Massive periarticular calcinosis of the soft tissues is a unique but not rare radiographic finding. On the contrary, tumoral calcinosis is a rare familial disease. Unfortunately, the term tumoral calcinosis has been liberally and imprecisely used to describe any massive collection of periarticular calcification, although this term actually refers to a hereditary condition associated with massive periarticular calcification. The inconsistent use of this term has created confusion throughout the literature. More important, if the radiologist is unfamiliar with tumoral calcinosis or disease processes that mimic this condition, then diagnosis could be impeded, treatment could be delayed, and undue alarm could be raised, possibly leading to unwarranted surgical procedures. The soft-tissue lesions of tumoral calcinosis are typically lobulated, well demarcated calcifications that are most often distributed along the extensor surfaces of large joints. There are many conditions with similar appearances, including the calcinosis of chronic renal failure, calcinosis universalis, calcinosis circumscripta, calcific tendonitis, synovial osteochondromatosis, synovial sarcoma, osteosarcoma, myositis ossificans, tophaceous gout, and calcific myonecrosis. The radiologist plays a critical role in avoiding unnecessary invasive procedures and in guiding the selection of appropriate tests that can result in a conclusive diagnosis of tumoral calcinosis. PMID- 16702461 TI - Multidetector CT for visualization of coronary stents. AB - Whereas the clinical diagnosis of in-stent thrombosis is straightforward, that of in-stent restenosis remains a problem, because although many patients experience chest pain after coronary stent placement, that symptom is secondary to ischemia in only a few. The use of a noninvasive technique to identify such patients for early invasive intervention versus more conservative management is thus highly desirable. Multidetector computed tomography (CT) performed with 16-section scanners recently emerged as such a technique and has overtaken modalities such as electron-beam CT and magnetic resonance imaging as an alternative to conventional angiography for the assessment of in-stent restenosis. The improved hardware design of the current 64-section CT scanners allows even better delineation of stent struts and lumen. The more reliable criterion of direct lumen visualization thus may be substituted for the presence of distal runoff, which lacks specificity for a determination of in-stent patency because of the possibility of collateral pathways. However, the capability to accurately visualize the in-stent lumen depends partly on knowledge of the causes of artifacts and how they can be compensated for with postprocessing and proper image display settings. In addition, an understanding of the major stent placement techniques used in the treatment of lesions at arterial bifurcations is helpful. PMID- 16702462 TI - Volume rendering versus maximum intensity projection in CT angiography: what works best, when, and why. AB - The introduction and widespread availability of 16-section multi-detector row computed tomographic (CT) technology and, more recently, 64-section scanners, has greatly advanced the role of CT angiography in clinical practice. CT angiography has become a key component of state-of-the-art imaging, with applications ranging from oncology (eg, staging of pancreatic or renal cancer) to classic vascular imaging (eg, evaluation of aortic aneurysms and renal artery stenoses) as well as newer techniques such as coronary artery imaging and peripheral runoff studies. With an average of 400-1000 images in each volume data set, three-dimensional postprocessing is crucial to volume visualization. Radiologists now have workstations that provide capabilities for evaluation of these data sets by using a range of software programs and processing tools. Although different systems have unique capabilities and functionality, all provide the options of volume rendering and maximum intensity projection for image display and analysis. These two postprocessing techniques have different advantages and disadvantages when used in clinical practice, and it is important that radiologists understand when and how each technique should be used. PMID- 16702463 TI - From the archives of the AFIP: Pleuropulmonary synovial sarcoma. AB - Pleuropulmonary synovial sarcoma (PPSS) is increasingly recognized as a subtype of sarcoma because of the recent identification of a distinctive chromosomal translocation specific to synovial sarcoma. Soft-tissue synovial sarcoma is far more common than PPSS and typically develops in para-articular locations of the extremities; affects young and middle-aged adults, with no difference in distribution between the sexes; and has well-documented radiologic manifestations. PPSS may arise in the chest wall, heart, mediastinum, pleura, or lung, and it shares patient demographics and several imaging features with its soft-tissue counterpart. Patients present with a cough, chest pain, or dyspnea. On chest radiographs, PPSS typically appears as a sharply marginated mass with uniform opacity, based either in the pleura or in the lung, and often accompanied by an ipsilateral pleural effusion. Computed tomographic images show a well circumscribed heterogeneously enhanced lesion without associated involvement of bone and without calcifications (except in the case of a chest wall primary tumor). Magnetic resonance imaging provides superior demonstration of nodular soft tissue and multilocular fluid-filled internal components of PPSS, in addition to peripheral rim enhancement after the intravenous administration of a gadolinium-based contrast material such as gadopentetate dimeglumine. Current treatment consists of surgical resection followed by chemotherapy, radiation therapy, or both. PMID- 16702464 TI - Best cases from the AFIP: Adenomyomatosis of the gallbladder. PMID- 16702465 TI - Best cases from the AFIP: Ulcerative colitis. PMID- 16702467 TI - Transvenous pacing leads and systemic thromboemboli in patients with intracardiac shunts: a multicenter study. AB - BACKGROUND: The risk of systemic thromboemboli associated with transvenous leads in the presence of an intracardiac shunt is currently unknown. METHODS AND RESULTS: To define this risk, we conducted a multicenter, retrospective cohort study of 202 patients with intracardiac shunts: Sixty-four had transvenous leads (group 1), 56 had epicardial leads (group 2), and 82 had right-to-left shunts but no pacemaker or implantable cardioverter defibrillator leads (group 3). Patient years were accrued until the occurrence of systemic thromboemboli or study termination. Censoring occurred in the event of complete shunt closure, death, or loss to follow-up. Mean ages for groups 1, 2, and 3 were 33.9+/-18.0, 22.2+/ 12.6, and 22.9+/-15.0 years, respectively. Respective oxygen saturations were 91.2+/-9.1%, 88.1+/-8.1%, and 79.7+/-6.7%. During respective median follow-ups of 7.3, 9.3, and 17.0 years, 24 patients had at least 1 systemic thromboembolus: 10 (15.6%), 5 (8.9%), and 9 (11.0%) in groups 1, 2, and 3, respectively. Univariate risk factors were older age (hazard ratio [HR], 1.05; P=0.0001), ongoing phlebotomy (HR, 3.1; P=0.0415), and an transvenous lead (HR, 2.4; P=0.0421). In multivariate, stepwise regression analyses, transvenous leads remained an independent predictor of systemic thromboemboli (HR, 2.6; P=0.0265). In patients with transvenous leads, independent risk factors were older age (HR, 1.05; P=0.0080), atrial fibrillation or flutter (HR, 6.7; P=0.0214), and ongoing phlebotomy (HR, 14.4; P=0.0349). Having had aspirin or warfarin prescribed was not protective. Epicardial leads were, however, associated with higher atrial (P=0.0407) and ventricular (P=0.0270) thresholds and shorter generator longevity (HR, 1.9; P=0.0176). CONCLUSIONS: Transvenous leads incur a >2-fold increased risk of systemic thromboemboli in patients with intracardiac shunts. PMID- 16702468 TI - Suppression of atherosclerotic plaque progression and instability by tissue inhibitor of metalloproteinase-2: involvement of macrophage migration and apoptosis. AB - BACKGROUND: Matrix metalloproteinase (MMP)-associated extracellular matrix degradation is thought to contribute to the progression and rupture of atherosclerotic plaques. However, direct evidence of this concept remains elusive. We hypothesized that overexpression of tissue inhibitor of metalloproteinase (TIMP)-1 or TIMP-2 would attenuate atherosclerotic plaque development and instability in high fat-fed apolipoprotein E-knockout (apoE(-/-)) mice. METHODS AND RESULTS: Seventy male apoE(-/-) mice (n=10/group) fed a high fat diet for 7 weeks were injected intravenously with first-generation adenoviruses expressing the gene for human TIMP-1 (RAdTIMP-1) or TIMP-2 (RAdTIMP 2) or a control adenovirus (RAd66) and were fed a high-fat diet for a further 4 weeks. Analysis of brachiocephalic artery plaques revealed that RAdTIMP-2 but not RAdTIMP-1 infection resulted in a marked reduction (48+/-13%, P<0.05) in lesion area compared with that in control animals. Markers associated with plaque instability, assessed by smooth muscle cell and macrophage content and the presence of buried fibrous caps, were significantly reduced by RAdTIMP-2. Effects on lesion size were not sustained with first-generation adenoviruses, but murine TIMP-2 overexpression mediated by helper-dependent adenoviral vectors exerted significant effects on plaques assessed 11 weeks after infection. In an attempt to determine the mechanism of action, we treated macrophages and macrophage derived foam cells with exogenous TIMP-2 in vitro. TIMP-2 significantly inhibited migration and apoptosis of macrophages and foam cells, whereas TIMP-1 failed to exert similar effects. CONCLUSIONS: Overexpression of TIMP-2 but not TIMP-1 inhibits atherosclerotic plaque development and destabilisation, possibly through modulation of macrophage and foam cell behavior. Helper-dependent adenovirus technology is required for these effects to be maintained long term. PMID- 16702469 TI - Impact of completeness of percutaneous coronary intervention revascularization on long-term outcomes in the stent era. AB - BACKGROUND: The importance of completeness of revascularization by percutaneous coronary intervention in patients with multivessel disease is unclear in that there is little information on the impact of incomplete revascularization outside of randomized trials. The objective of this study is to compare long-term mortality and subsequent revascularization for percutaneous coronary intervention patients receiving stents who were completely revascularized (CR) with those who were incompletely revascularized (IR). METHODS AND RESULTS: Patients from New York State's Percutaneous Coronary Interventions Reporting System were subdivided into patients who were CR and IR. Then subsets of IR patients were contrasted with CR patients. Differences in long-term survival and subsequent revascularization for CR and IR patients were compared after adjustment for differences in preprocedural risk. A total of 68.9% of all stent patients with multivessel disease who were studied were IR, and 30.1% of all patients had total occlusions and/or > or =2 IR vessels. At baseline, the following patients were at higher risk: those who were older and those with more comorbid conditions, worse ejection fraction, and more renal disease and stroke. After adjustment for these baseline differences, IR patients were significantly more likely to die at any time (adjusted hazard ratio=1.15; 95% confidence interval, 1.01 to 1.30) than CR patients. IR patients with total occlusions and a total of > or =2 IR vessels were at the highest risk compared with CR patients (hazard ratio=1.36; 95% confidence interval, 1.12 to 1.66). CONCLUSIONS: IR with stenting is associated with an adverse impact on long-term mortality, and consideration should be given to either achieving CR, opting for surgery, or monitoring percutaneous coronary intervention patients with IR more closely after discharge. PMID- 16702470 TI - Focal pharmacological modulation of atrioventricular nodal conduction via implantable catheter: a novel therapy for atrial fibrillation? AB - BACKGROUND: Pharmacological ventricular rate control is an acceptable atrial fibrillation (AF) therapy limited by systemic toxicity. We postulate that focal catheter-based drug delivery into the atrioventricular nodal (AVN) region may effectively control ventricular rate during AF without systemic toxicity. This study evaluated the effects of focally administered acetylcholine on AVN conduction and refractoriness during sinus rhythm and AF. METHODS AND RESULTS: Canines (n=7) were anesthetized and instrumented to assess cardiac electrophysiology and blood pressure. A custom drug delivery catheter was implanted in the AVN region. Incremental doses of acetylcholine starting at 10 microg/min were infused until complete AV block was achieved. Acetylcholine induced dose-dependent AV block. AF induction and electrophysiology measurements were performed during baseline and acetylcholine-induced first-degree and third degree AV block. During AF, infusion of acetylcholine decreased ventricular rates from 182+/-32 to 77+/-28 and 28+/-8 bpm (first-degree and third-degree AV block, respectively; P<0.05). At the first-degree AV block dose, AVN effective refractory period increased from 186+/-37 to 282+/-33 ms, and Wenckebach cycle length increased from 271+/-29 to 378+/-58 ms (P<0.05). The first-degree AV block dose prolonged AV and AH intervals by 26% and 23% (P<0.05), whereas AA intervals and blood pressure remained unchanged, demonstrating a local effect. All effects were reversed 20 minutes after infusion was stopped. CONCLUSIONS: Focal acetylcholine delivery into the AVN increased AVN refractoriness and significantly decreased ventricular rate response during induced AF in a dose related, reversible manner without systemic side effects. This may represent a novel therapy for AF whereby ventricular rate is controlled with the use of an implantable drug delivery system. PMID- 16702472 TI - Effects of conjugated equine estrogen on stroke in the Women's Health Initiative. AB - BACKGROUND: The Women's Health Initiative (WHI) Estrogen Alone trial assessed the balance of benefits and risks of hormone use in healthy postmenopausal women. The trial was stopped prematurely because there was no benefit for coronary heart disease and an increased risk of stroke. This report provides a thorough analysis of the stroke finding using the final results from the completed trial database. METHODS AND RESULTS: The WHI Estrogen Alone hormone trial is a multicenter, double-blind, placebo-controlled, randomized clinical trial in 10,739 women aged 50 to 79 years who were given daily conjugated equine estrogen (CEE; 0.625 mg; n=5310) or placebo (n=5429). During an average follow-up of 7.1 years, there were 168 strokes in the CEE group and 127 in the placebo group; 80.3% of strokes were ischemic. For all stroke the intention-to-treat hazard ratio [HR] (95% CI) for CEE versus placebo was 1.37 (1.09 to 1.73). The HR (95% CI) was 1.55 (1.19 to 2.01) for ischemic stroke and 0.64 (0.35, 1.18) for hemorrhagic stroke. The HRs indicate excess risk of ischemic stroke was apparent in all categories of baseline stroke risk, including younger and more recently menopausal women and in women with prior or current use of statins or aspirin. CONCLUSIONS: CEE increases the risk of ischemic stroke in generally healthy postmenopausal women. The excess risk appeared to be present in all subgroups of women examined, including younger and more recently menopausal women. There was no convincing evidence to suggest that CEE had an effect on the risk of hemorrhagic stroke. PMID- 16702471 TI - Topical sonic hedgehog gene therapy accelerates wound healing in diabetes by enhancing endothelial progenitor cell-mediated microvascular remodeling. AB - BACKGROUND: Sonic hedgehog (Shh) is a prototypical morphogen known to regulate epithelial-mesenchymal interaction during embryonic development. Recent observations indicate that exogenous administration of Shh can induce angiogenesis and may accelerate repair of ischemic myocardium and skeletal muscle. Because angiogenesis plays a pivotal role in wound repair, we hypothesized that activation of the hedgehog pathway may promote a favorable effect on microvascular remodeling during cutaneous wound healing and thereby accelerate wound closure. Because diabetes is associated with impaired wound healing, we tested this hypothesis in a diabetic model of cutaneous wound repair. METHODS AND RESULTS: In Ptc1-LacZ mice, cutaneous injury resulted in LacZ expression, indicating that expression of the Shh receptor Patched was induced and therefore that the Shh signaling pathway was intact postnatally and upregulated in the process of wound repair. In diabetic mice, topical gene therapy with the use of naked DNA encoding for Shh resulted in significant local gene expression and acceleration of wound recovery. The acceleration in wound healing was notable for increased wound vascularity. In bone marrow transplantation models, the enhanced vascularity of the wound was shown to be mediated, at least in part, by enhanced recruitment of bone marrow-derived endothelial progenitor cells. In vitro, Shh promoted production of angiogenic cytokines from fibroblasts as well as proliferation of dermal fibroblasts. Furthermore, Shh directly promoted endothelial progenitor cell proliferation, migration, adhesion, and tube formation. CONCLUSIONS: These findings suggest that a simple strategy of topically applied Shh gene therapy may have significant therapeutic potential for enhanced wound healing in patients with impaired microcirculation such as occurs in diabetes. PMID- 16702473 TI - High-resolution quantitative computed tomography demonstrating selective enhancement of medium-size collaterals by placental growth factor-1 in the mouse ischemic hindlimb. AB - BACKGROUND: The process of arteriogenesis after occlusion of a major artery is poorly understood. We have used high-resolution microcomputed tomography (mu-CT) imaging to define the arteriogenic response in the mouse model of hindlimb ischemia and to examine the effect of placental growth factor-1 (PlGF-1) on this process. METHODS AND RESULTS: After common femoral artery ligation, mu-CT imaging demonstrated formation of collateral vessels originating near the ligation site in the upper limb and connecting to the ischemic calf muscle region. Three dimensional mu-CT and quantitative image analysis revealed changes in the number of segments and the segmental volume of vessels, ranging from 8 to 160 microm in diameter. The medium-size vessels (48 to 160 microm) comprising 85% of the vascular volume were the major contributor (188%) to the change in vascular volume in response to ischemia. Intramuscular injections of Ad-PlGF-1 significantly increased Sca1+ cells in the circulation, alpha-actin-stained vessels, and perfusion of the ischemic hindlimb. These effects were predominantly associated with an increase in vascular volume contributed by the medium-size (96 to 144 microm) vessels as determined by mu-CT. CONCLUSIONS: High-resolution mu-CT delineated the formation of medium-size collaterals representing a major vascular change that contributed to the restoration of vascular volume after ischemia. This effect is selectively potentiated by PlGF-1. Such selective enhancement of arteriogenesis by therapeutically administered PlGF-1 demonstrates a desirable biological activity for promoting the growth of functionally relevant vasculature. PMID- 16702474 TI - Implementation of guidelines improves the standard of care: the Viennese registry on reperfusion strategies in ST-elevation myocardial infarction (Vienna STEMI registry). AB - BACKGROUND: The purpose of this study was to determine whether implementation of recent guidelines improves in-hospital mortality from acute ST-elevation myocardial infarction (STEMI) in a metropolitan area. METHODS AND RESULTS: We organized a network that consisted of the Viennese Ambulance Systems, which is responsible for diagnosis and triage of patients with acute STEMI, and 5 high volume interventional cardiology departments to expand the performance of primary percutaneous catheter intervention (PPCI) and to use the fastest available reperfusion strategy in STEMI of short duration (2 to 3 hours from onset of symptoms), either PPCI or thrombolytic therapy (TT; prehospital or in-hospital), respectively. Implementation of guidelines resulted in increased numbers of patients receiving 1 of the 2 reperfusion strategies (from 66% to 86.6%). Accordingly, the proportion of patients not receiving reperfusion therapy dropped from 34% to 13.4%, respectively. PPCI usage increased from 16% to almost 60%, whereas the use of TT decreased from 50.5% to 26.7% in the participating centers. As a consequence, in-hospital mortality decreased from 16% before establishment of the network to 9.5%, including patients not receiving reperfusion therapy. Whereas PPCI and TT demonstrated comparable in-hospital mortality rates when initiated within 2 to 3 hours from onset of symptoms, PPCI was more effective in acute STEMI of >3 but <12 hours' duration. CONCLUSIONS: Implementation of recent guidelines for the treatment of acute STEMI by the organization of a cooperating network within a large metropolitan area was associated with a significant improvement in clinical outcomes. PMID- 16702475 TI - Electrocardiogram: still the cardiologist's best friend. PMID- 16702476 TI - Images in cardiovascular medicine. Complete ectopia cordis. PMID- 16702477 TI - Images in cardiovascular medicine. Left ventricular hypertrophy and outflow tract obstruction in a patient with anorexia nervosa. PMID- 16702478 TI - Images in cardiovascular medicine. Takotsubo cardiomyopathy. PMID- 16702480 TI - Letter regarding article by Tomic et al, "transcriptomic and proteomic patterns of systemic inflammation in on-pump and off-pump coronary artery bypass grafting". PMID- 16702483 TI - Viewpoint: how cardiac imaging has developed. PMID- 16702484 TI - Pushing drug-eluting stents into uncharted territory: simpler than you think- more complex than you imagine. PMID- 16702485 TI - Pediatric mechanical circulatory support: challenges and opportunities. PMID- 16702486 TI - Percutaneous mitral valve repair: a fertile field of innovative treatment strategies. PMID- 16702487 TI - Outcomes of children bridged to heart transplantation with ventricular assist devices: a multi-institutional study. AB - BACKGROUND: Current ventricular assist devices (VADs) in the United States are designed primarily for adult use. Data on VADs as a bridge to transplantation in children are limited. METHODS AND RESULTS: A multi-institutional, prospectively maintained database of outcomes in children after listing for heart transplantation (n=2375) was used to analyze outcomes of VAD patients (n=99, 4%) listed between January 1993 and December 2003. Median age at VAD implantation was 13.3 years (range, 2 days to 17.9 years); diagnoses were cardiomyopathy (78%) and congenital heart disease (22%). Mean duration of support was 57 days (range, 1 to 465 days). Seventy-three percent were supported with a long-term device, with 39% requiring biventricular support. Seventy-seven patients (77%) survived to transplantation, 5 patients were successfully weaned from support and recovered, and 17 patients (17%) died on support. In the recent era (2000 to 2003), successful bridge to transplantation with VAD was achieved in 86% of patients. Peak hazard for death while waiting was the first 2 weeks after VAD placement. Risk factors for death while awaiting a transplant included earlier era of implantation (P=0.05), female gender (P=0.02), and congenital disease diagnosis (P=0.05). There was no difference in 5-year survival after transplantation for patients on VAD at time of transplantation as compared with those not requiring VAD. CONCLUSIONS: VAD support in children successfully bridged 77% of patients to transplantation, with posttransplantation outcomes comparable to those not requiring VAD. These encouraging results emphasize the need to further understand patient selection and to delineate the impact of VAD technology for children. PMID- 16702488 TI - Biomarkers of cardiovascular disease: molecular basis and practical considerations. PMID- 16702489 TI - AHA/ACC guidelines for secondary prevention for patients with coronary and other atherosclerotic vascular disease: 2006 update: endorsed by the National Heart, Lung, and Blood Institute. PMID- 16702490 TI - Brachial pressure-independent reduction in carotid stiffness after long-term angiotensin-converting enzyme inhibition in diabetic hypertensives. AB - Hypertension and diabetes are associated with an increased arterial stiffness. A direct blood pressure-independent effect of angiotensin-converting enzyme inhibitors on arterial stiffness has never been unequivocally demonstrated. In this mechanistic study, we used an experimental design in which patients responding to 1 month treatment with 4 mg perindopril were randomized double blind to either 4 mg perindopril or 8 mg perindopril for 6 months. We determined carotid distensibility with echotracking and applanation tonometry at baseline and after the 7-month treatment period in 57 essential hypertensive patients with type 2 diabetes (age 63+/-7 years). We monitored ambulatory blood pressure at baseline and after treatment. After 7 months treatment, 24-hour ambulatory blood pressure significantly decreased, with no significant difference between 4 mg and 8 mg perindopril. Carotid distensibility increased more after 8 mg perindopril compared with 4 mg perindopril (8 mg: from 13.1+/-5.9 to 16.0+/-6.7 kPa(-1)x10( 3); 4 mg: from 13.2+/-5.2 to 12.7+/-5.9 kPa(-1)x10(-3); ANOVA, dose-period interaction, P<0.05). Carotid internal diameter and elastic modulus were significantly lower after 8 mg perindopril compared with 4 mg perindopril, independent of blood pressure reduction. These results indicate a dose-dependent and blood pressure-independent reduction in carotid stiffness under chronic treatment with an angiotensin-converting enzyme inhibitor. They suggest that arterial distensibility was increased through an inward remodeling, leading to a reduction in wall stress, thus reducing elastic modulus. They also suggest that long-term administration of high doses (8 mg) of perindopril is required to improve carotid structure and function in hypertensive patients with type 2 diabetes. PMID- 16702491 TI - Cardiovascular function in a rat model of diet-induced obesity. AB - The obesity-prone/obesity-resistant rat model has been used to study mechanisms responsible for obesity-related abnormalities in renal function and blood pressure, but whether this model exhibits cardiac dysfunction has not been determined. We tested the hypothesis that obesity-prone rats would display cardiovascular abnormalities seen in other diet-induced obese models (ie, hypertension, tachycardia, left ventricular hypertrophy, increased collagen deposition, reduced cardiac contractility, and increased end diastolic pressure). Male Sprague-Dawley rats were fed a control diet or a moderate fat diet containing 32% kcal as fat while hemodynamics were continuously monitored using telemetry. After 12 weeks, obesity-prone rats were significantly heavier and had greater body fat compared with obesity-resistant rats and controls, but daily (20 hours/d) averages and diurnal rhythms of blood pressure and heart rate did not differ among groups. Echocardiographic indices of cardiac structure and function, histological evidence of cardiac collagen, and directly measured heart weights did not differ among groups. Peak left ventricular pressure, end diastolic pressure, +dP/dt, and -dP/dt were also not significantly different among groups. Plasma cholesterol and hepatic cholesterol were significantly higher in obesity prone rats compared with obesity-resistant rats and controls; hepatic triglycerides were higher in obesity-prone rats compared with controls (P< or =0.05). Leptin was significantly higher in obesity-prone rats compared with controls and across all groups was significantly correlated with body fat (P< or =0.05). These results suggest that 12 weeks of a moderate fat diet in the obesity prone/obesity-resistant rat model induced lipid and endocrine abnormalities typical of obesity but was not sufficient to cause significant cardiac abnormalities. PMID- 16702492 TI - Successful treatment of severe atopic dermatitis in a child and an adult with the T-cell modulator efalizumab. PMID- 16702493 TI - Development of atopic dermatitis during the first 3 years of life: the Copenhagen prospective study on asthma in childhood cohort study in high-risk children. AB - OBJECTIVES: To describe the development of atopic dermatitis (AD) during the first 3 years of life and identify the localization of the early skin lesions that predicts the development of AD. DESIGN: Prospective, longitudinal, birth cohort study of children born to mothers with a history of asthma, followed up for 3 years with scheduled visits every 6 months as well as visits for onset or acute exacerbations of skin symptoms. SETTING: The cohort was recruited from greater Copenhagen, Denmark, and followed up at a clinical research unit, which controlled all diagnoses and treatment of skin diseases. PARTICIPANTS: A total of 411 infants were enrolled in the cohort; 55 had incomplete follow-up and were excluded from certain analyses. MAIN OUTCOME MEASURES: Atopic dermatitis was defined based on the criteria of Hanifin and Rajka, and severity was assessed by the SCORAD (Scoring Atopic Dermatitis) index. Predictive odds ratios of early skin lesions for those who developed AD vs those who did not were calculated. RESULTS: The cumulative incidence of AD by age 3 years was 44% (155/356). The prevalence rate peaked at age 2 years for boys and at age 2.5 years for girls, but there were no other sex differences in the proportion of children developing AD. Skin involvement in infants with AD was found to begin at the scalp, forehead, ear, and neck in a balaclava-like pattern and continue to the extensor sides and trunk, finally affecting the flexor sides of the extremities. Early skin lesions of arms and joints best predicted AD at age 3 years. CONCLUSIONS: Atopic dermatitis begins at the scalp, forehead, ear, and neck in a balaclava like pattern. Eczema at the arms and joints provides the highest predictive value for the development of AD at age 3 years. This may be used for early prediction and intervention of AD. PMID- 16702494 TI - Randomized controlled trial of adjuvant oral dexamethasone pulse therapy in pemphigus vulgaris: PEMPULS trial. AB - OBJECTIVE: To determine the therapeutic effect of adjuvant dexamethasone pulse therapy when given in addition to conventional treatment of pemphigus vulgaris. DESIGN: A randomized, placebo-controlled trial. SETTING: International European, multicenter outpatient and inpatient study. PATIENTS: Of the 20 enrolled patients, 11 were randomized to the dexamethasone pulse (DP) group and 9 to the placebo pulse (PP) group. INTERVENTIONS: Oral dexamethasone in 300-mg pulses or PPs 3 days per month. During the intervention, the DP and PP groups received conventional treatment with prednisolone, 80 mg/d, which was tapered across 19 weeks, and azathioprine sodium, 3 mg/kg per day, until the end of the study. Monthly pulses were continued until prednisolone treatment was tapered to 0 mg. MAIN OUTCOME MEASURES: Number of patients in remission, time to and duration of remission, cumulative prednisolone dose, and occurrence of adverse events during 1 year of follow-up. RESULTS: Eight of the 11 DP-treated patients and all 9 PP treated patients achieved remission. Mean time to remission was 173 days with DP and 176 days with PP. The mean duration of remission within the first year was 151 days for DP and 141 days for PP. Mean cumulative prednisolone dose was 5300 mg for DP and 4882 mg for PP. Weight gain (>5% of baseline) occurred in 8 DP treated patients compared with 1 PP-treated patient (P<.01). We found no statistically significant difference (P>.05) of an adjuvant effect of DP on remission of pemphigus vulgaris. CONCLUSION: In patients with new pemphigus vulgaris disease activity, there was no benefit of oral DP therapy given in addition to conventional treatment. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00127764. PMID- 16702495 TI - Staging accuracy in mycosis fungoides and sezary syndrome using integrated positron emission tomography and computed tomography. AB - OBJECTIVES: To evaluate the usefulness of integrated positron emission tomography and computed tomography (PET/CT) in staging mycosis fungoides (MF) and Sezary syndrome and to correlate PET/CT data with histopathologic diagnosis of lymph nodes (LNs). DESIGN: A single-center, prospective cohort analysis. SETTING: Academic referral center for cutaneous lymphoma. PATIENTS: Thirteen patients with MF and SS at risk for secondary LN involvement. Interventions Patients were clinically evaluated based on general physical examination, total body skin examination, and laboratory screening. They underwent integrated PET/CT followed by excisional biopsy of LNs. MAIN OUTCOME MEASURES: We used PET/CT to assess LN size and metabolic activity. Enlarged LNs were defined as axillary or inguinal LNs with a short axis 1.5 cm or larger; or cervical LN, with a short axis 1.0 cm or larger. We classified LN pathologic results according to National Cancer Institute (LN1-4) and World Health Organization (WHO 1-3) criteria. We quantified PET activity using standardized uptake value (SUV) and correlated with LN grade. RESULTS: Based on CT size criteria alone, only 5 patients had enlarged LNs, whereas PET revealed hypermetabolic LNs in all 13 patients. Six patients had LN1 3, and 7 had effacement of LN architecture by lymphoma cells (LN4). Of the 7 patients with LN4 nodes, 4 had SS, and 3 had tumorous MF. Two patients with LN4 nodes had inguinal LNs smaller than 1.5 cm and would have been assigned an N0 classification without the use of integrated PET/CT. Correlation of SUV with LN grade revealed that LN1-3 nodes were associated with a mean SUV of 2.7 (median SUV, 2.2; range, 2.0-4.7) and LN4 nodes were associated with a mean SUV of 5.4 (median SUV, 3.9; range, 2.1-11.8). Patients with large cell transformation had the highest SUVs. CONCLUSIONS: For staging MF and SS, PET/CT was more sensitive in detecting LN involved by lymphoma compared with CT data alone and thus may provide more accurate staging and prognostic information. The intensity of PET activity correlated with histologic LN grade. PMID- 16702496 TI - Pathogenic link between hydroa vacciniforme and Epstein-Barr virus-associated hematologic disorders. AB - OBJECTIVES: To determine the pathogenic association of latent Epstein-Barr virus (EBV) infections with both typical hydroa vacciniforme (HV) and severe HV-like eruptions, and to survey the complications and outcomes of patients. DESIGN: Case series. PATIENTS: Twenty-nine patients with HV or severe HV-like eruptions. INTERVENTIONS: In situ hybridization and immunostaining of biopsy specimens; extraction of DNA samples from cutaneous lesions and/or peripheral blood mononuclear cells for EBV DNA assay. MAIN OUTCOME MEASURES: Clinicopathologic manifestations, hematologic findings, complications, and outcomes; presence of latent EBV infection. RESULTS: T cells positive for EBV-encoded small nuclear RNA (EBER) were detected, to various degrees, in cutaneous infiltrates in 28 (97%) of 29 patients, including all 6 patients with definite HV with a positive phototest reaction, 11 of 12 patients with probable HV without evidence of photosensitivity, and all 11 patients with severe HV associated with systemic symptoms. In addition to EBER-positive T cells, many cytotoxic T lymphocytes expressing T-cell intracellular antigen 1 and granzyme B were present in the cutaneous lesions. Natural killer (NK) cells were absent or at a background level. The UV-induced cutaneous lesions showed histopathologic findings consistent with those of HV, containing many EBER-positive cells. Although no hematologic abnormalities were found in the definite and probable HV groups, the amounts of EBV DNA were increased in the peripheral blood mononuclear cells. By contrast, the severe HV group had markedly increased levels of EBV DNA associated with NK-cell lymphocytosis, and complications including chronic active EBV infection, hypersensitivity to mosquito bites, and hemophagocytic syndrome. Five patients with severe disease died of EBV-associated NK/T-cell lymphomas or hemophagocytic syndrome 2 to 14 years after onset. CONCLUSION: Both typical and severe HV are included within the spectrum of cutaneous disorders mediated by EBV infected T cells, and the severe HV group may have overt EBV-associated NK/T-cell lymphoproliferative disorders with a frequently fatal outcome. PMID- 16702497 TI - Adapalene gel, 0.1%, as maintenance therapy for acne vulgaris: a randomized, controlled, investigator-blind follow-up of a recent combination study. AB - OBJECTIVE: To assess the maintenance effect of adapalene gel, 0.1%, relative to gel vehicle in subjects successfully treated in a previous 12-week study of adapalene-doxycycline, 100 mg, combination therapy. DESIGN: Multicenter, investigator-blind, randomized, controlled study. SETTING: Thirty-four US centers. SUBJECTS: A total of 253 subjects with severe acne vulgaris who showed at least moderate improvement from baseline (50% improvement from baseline) when treated with either adapalene plus doxycycline or doxycycline plus gel vehicle in a previous 12-week study. INTERVENTIONS: Subjects were randomized to receive adapalene gel, 0.1%, or gel vehicle once daily for 16 weeks. MAIN OUTCOME MEASURES: Efficacy and safety criteria included maintenance rate (subjects maintaining at least 50% improvement in lesion counts from previous therapy), lesion counts (total, inflammatory, and noninflammatory), global severity assessment, cutaneous tolerability, and adverse events. RESULTS: Adapalene maintenance therapy resulted in significantly larger maintenance rates (75% vs 54%; P<.001) and significantly lower lesion counts (total [P = .005], inflammatory [P = .01], and noninflammatory [P = .02]) compared with gel vehicle. Adapalene was safe and well tolerated in this study. Conclusion This study demonstrates a clinical benefit of continued treatment with adapalene gel, 0.1%, as a maintenance therapy for acne. PMID- 16702498 TI - Comparison of tazarotene and minocycline maintenance therapies in acne vulgaris: a multicenter, double-blind, randomized, parallel-group study. AB - OBJECTIVE: To evaluate the efficacy of 3 maintenance regimens (topical tazarotene, oral minocycline hydrochloride, or both) in sustaining improvement in acne. DESIGN: Multicenter, open-label treatment phase followed by double-blind, randomized, parallel-group maintenance phase. SETTING: Ambulatory patients in research or referral centers. PATIENTS: Volunteer sample of 189 patients with moderately severe to severe acne vulgaris (110 entered maintenance phase, 90 completed, and 2 discontinued because of adverse events). INTERVENTIONS: All patients were treated with 0.1% tazarotene gel (each evening) and a 100-mg capsule (twice daily) of minocycline hydrochloride for up to 12 weeks. Patients with 75% or greater global improvement at week 12 were randomly assigned to 12 weeks of maintenance therapy with tazarotene gel plus placebo capsules, vehicle gel plus minocycline capsules, or tazarotene gel plus minocycline capsules. MAIN OUTCOME MEASURES: Overall disease severity, global improvement, and lesion counts. RESULTS: All regimens were effective in sustaining improvements in acne. After 12 weeks of maintenance therapy, the mean reductions from baseline in noninflammatory and inflammatory lesion count, respectively, were 60% and 54% with tazarotene, 52% and 66% with minocycline, and 64% and 66% with tazarotene plus minocycline. At week 24, more than 80% of patients in each group had maintained a 50% or greater global improvement from baseline, and more than 50% had maintained a 75% or greater global improvement. CONCLUSIONS: A high percentage of patients with moderately severe to severe acne can maintain improvement in their condition with topical retinoid monotherapy. Maintenance with combination tazarotene and minocycline therapy showed a trend for greater efficacy but no statistical significance vs tazarotene alone. Topical retinoid monotherapy should be considered for maintenance to help minimize antibiotic exposure. PMID- 16702499 TI - Angiokeratoma corporis diffusum in a patient with no recognizable enzyme abnormalities. AB - BACKGROUND: Angiokeratoma corporis diffusum is a clinical variant of angiokeratoma that is typically associated with an enzyme deficiency in the metabolism of glycoprotein, most notably Fabry disease, resulting in many other systemic manifestations. OBSERVATIONS: We report a case of angiokeratoma corporis diffusum that did not have an identifiable enzyme deficiency. A review of the literature revealed few similar cases. CONCLUSION: Angiokeratoma corporis diffusum without recognizable enzyme deficiencies appears to be a distinct clinical entity with a benign course. PMID- 16702500 TI - Kindler syndrome: a new mutation and new diagnostic possibilities. AB - BACKGROUND: Kindler syndrome (KS) is a rare genetic disorder that is characterized by blistering in infancy, followed by the onset of poikiloderma and photosensitivity in childhood. The recently elucidated molecular pathogenesis involves mutations in KIND1, a gene encoding the protein kindlin-1, which is involved in the attachment of the actin cytoskeleton to the extracellular matrix in basal keratinocytes. OBSERVATIONS: We describe a child with the neonatal diagnosis of epidermolysis bullosa simplex who developed poikiloderma and skin fragility at 6 years of age. His skin showed diminished staining with anti kindlin-1 antibody, and genetic analysis revealed that he was a compound heterozygote with a previously unreported mutation in KIND1. Ultrastructural clues to the diagnosis of KS were present in a biopsy specimen that was obtained when the patient was 10 months old, before he developed poikiloderma and photosensitivity. CONCLUSIONS: In this case, a combination of a known mutation (R271X) and a newly described mutation (1755delT) in the KIND1 gene produced loss of function in kindlin-1, leading to the clinical features of KS. Ultrastructural findings characteristic of KS were evident years before the onset of poikiloderma and sun sensitivity. In infancy, electron microscopy can enable early, accurate diagnosis of KS. PMID- 16702501 TI - Mucocutaneous neuromas: an underrecognized manifestation of PTEN hamartoma-tumor syndrome. AB - BACKGROUND: The spectrum of clinical findings associated with PTEN tumor suppressor gene germline mutations, referred to as PTEN hamartoma-tumor syndrome (PHTS), includes Cowden and Bannayan-Riley-Ruvalcaba syndromes. Although the skin is the ectodermal structure most often affected by these autosomal dominant genodermatoses, abnormalities of neural tissues are frequently observed. OBSERVATIONS: We describe a 5-year-old boy with macrocephaly, prominent corneal nerves, and progressive development of multiple painful, dome-shaped, translucent pink to skin-colored papules on the vermilion portion of the upper lip, fingers, palms, and shins. Histologic evaluation demonstrated dermal proliferation of well demarcated nerve bundles associated with abundant mucin and surrounded by a distinct perineural sheath, findings diagnostic of a nonencapsulated neuroma. Genetic analysis revealed a novel heterozygous germline nonsense mutation in PTEN, predicted to result in a truncated PTEN protein. To our knowledge, this represents the first report of multiple neuromas as the sole mucocutaneous manifestation of PHTS. CONCLUSIONS: This article highlights neuromas as a cutaneous sign of PHTS, drawing attention to manifestations of PHTS in neural tissues of the skin, eye, gastrointestinal tract, and brain. Along with multiple endocrine neoplasia type 2B, PHTS should be considered in the differential diagnosis of multiple mucocutaneous neuromas, particularly those involving extrafacial sites. PMID- 16702502 TI - Topical calcineurin inhibitors for atopic dermatitis: balancing clinical benefit and possible risks. PMID- 16702503 TI - Acne maintenance therapy: expanding the role of topical retinoids? PMID- 16702504 TI - A slowly enlarging, unilateral, erythematous macular lesion. PMID- 16702505 TI - Generalized maculopapules with fever and cervical lymphadenopathy. PMID- 16702506 TI - Abdominal plaque in a 10-day-old boy. PMID- 16702508 TI - Phototesting in patients with Smith-Lemli-Opitz syndrome confirms sensitivity to UV-A. PMID- 16702507 TI - Multiple violaceous nodules in a 46-year-old man. PMID- 16702509 TI - Patient perceptions about a novel form of patient-assisted teledermatology. PMID- 16702510 TI - Peginterferon alfa-2b for mycosis fungoides. PMID- 16702511 TI - Oral involvement in hydroa vacciniforme. PMID- 16702512 TI - Follicular hyperkeratosis, hemorrhage, and corkscrew hair. PMID- 16702513 TI - Department of Surgery, New York Presbyterian Hospital-Weill Cornell Medical Center. PMID- 16702514 TI - Lessons. PMID- 16702515 TI - Result of a national audit of bariatric surgery performed at academic centers: a 2004 University HealthSystem Consortium Benchmarking Project. AB - HYPOTHESIS: Bariatric surgery performed at US academic centers is safe and associated with low mortality. DESIGN: Multi-institutional consecutive cohort study. SETTING: Academic medical centers. PATIENTS AND INTERVENTIONS: We audited the medical records from 40 consecutive bariatric surgery cases performed between October 1, 2003, and March 31, 2004, at each of the 29 institutions participating in the University HealthSystem Consortium Bariatric Surgery Benchmarking Project. All medical records that met inclusion criteria (patient age, >17 and <65 years; and body mass index [calculated as weight in kilograms divided by the square of height in meters], 35-70) and exclusion criteria (previous bariatric surgery) were reviewed and data were collected on a standardized form. MAIN OUTCOME MEASURES: Demographic data, operative time, blood loss, transfusion requirement, complications, readmission, reoperation, and in-hospital and 30-day mortality. RESULTS: Data from 1144 bariatric surgery cases were reviewed from 29 University HealthSystem Consortium institutions. The specific bariatric procedures included gastric bypass (91.7%), gastroplasty or gastric banding (8.2%), and biliopancreatic diversion (0.1%). For gastric bypass procedures (n = 1049), the mean patient age was 43 years and mean body mass index was 49; 76% of procedures were performed laparoscopically, with a conversion rate of 2.2%; the overall complication rate was 16%, with an anastomotic leakage rate of 1.6%; the 30-day readmission rate was 6.6%; and the 30-day mortality rate was 0.4%. For restrictive procedures (n = 94), the mean patient age was 45 years and mean body mass index was 45; 92% of procedures were performed laparoscopically with no conversion; the overall complication rate was 3.2%; the 30-day readmission rate was 4.3%; and the 30-day mortality rate was 0%. CONCLUSIONS: Within the context of the 2004 University HealthSystem Consortium Bariatric Surgery Benchmarking Project, the risk for death within 30 days after bariatric surgery at academic centers is less than 1%. In addition, the practice of bariatric surgery at these centers has shifted from open surgery to predominately laparoscopic surgery. These quality-controlled outcome data can be used as a benchmark for the practice of bariatric surgery at most US hospitals. PMID- 16702516 TI - Risk factors for hepatic morbidity following nonoperative management: multicenter study. AB - HYPOTHESIS: Early risk factors for hepatic-related morbidity in patients undergoing initial nonoperative management of complex blunt hepatic injuries can be accurately identified. DESIGN: Multicenter historical cohort. SETTING: Seven urban level I trauma centers. PATIENTS: Patients from January 2000 through May 2003 with complex (grades 3-5) blunt hepatic injuries not requiring laparotomy in the first 24 hours. INTERVENTION: Nonoperative treatment of complex blunt hepatic injuries. MAIN OUTCOME MEASURES: Complications and treatment strategies. RESULTS: Of 699 patients with complex blunt hepatic injuries, 453 (65%) were treated nonoperatively. Overall, 61 patients (13%) developed 87 hepatic complications including bleeding (38), biliary (bile peritonitis, 7; bile leak, 9; biloma, 11; biliary-venous fistula, 1; and bile duct injury, 1), abdominal compartment syndrome (5), and infections (abscess, 7; necrosis, 2; and suspected abdominal sepsis, 6), which required 86 multimodality treatments (angioembolization, 32; endoscopic retrograde cholangiopancreatography and stenting, 9; interventional radiology drainage, 16; paracentesis, 1; laparotomy, 24; and laparoscopy, 4). Hepatic complications developed in 5% (13 of 264) of patients with grade 3 injuries, 22% (36 of 166) of patients with grade 4 injuries, and 52% (12 of 23) of patients with grade 5 injuries. Univariate analysis revealed 24-hour crystalloid, total and first 24-hour packed red blood cells, fresh frozen plasma, platelet, and cryoprecipitate requirements and liver injury grade to be significant but only liver injury grade (grade 4 odds ratio, 4.439; grade 5 odds ratio, 12.001) and 24-hour transfusion requirement (odds ratio, 6.446) predicted complications by multivariable analysis. CONCLUSIONS: Nonoperative management of high-grade liver injuries is associated with significant morbidity and correlates with grade of liver injury. Screening patients with transfusion requirements and high-grade injuries may result in earlier diagnosis and treatment of hepatic related complications. PMID- 16702517 TI - Solitary colorectal liver metastasis: resection determines outcome. AB - BACKGROUND: Hepatic resection (HR) and radiofrequency ablation (RFA) have been proposed as equivalent treatments for colorectal liver metastasis. HYPOTHESIS: Recurrence patterns after HR and RFA for solitary liver metastasis are similar. DESIGN: Analysis of a prospective database at a tertiary care center with systematic review of follow-up imaging in all of the patients. PATIENTS AND METHODS: Patients with solitary liver metastasis as the first site of metastasis treated for cure by HR or RFA were studied (patients received no prior liver directed therapy). Prognostic factors, recurrence patterns, and survival rates were analyzed. RESULTS: Of the 180 patients who were studied, 150 underwent HR and 30 underwent RFA. Radiofrequency ablation was used when resection would leave an inadequate liver remnant (20 patients) or comorbidity precluded safe HR (10 patients). Tumor size and treatment determined recurrence and survival. The local recurrence (LR) rate was markedly lower after HR (5%) than after RFA (37%) (P<.001). Treatment by HR was associated with longer 5-year survival rates than RFA, including LR-free (92% vs 60%, respectively; P<.001), disease-free (50% vs 0%, respectively; P = .001), and overall (71% vs 27%, respectively; P<.001) survival rates. In the subset with tumors 3 cm or larger (n = 79), LR occurred more frequently following RFA (31%) than after HR (3%) (P = .001), with a 5-year LR-free survival rate of 66% after RFA vs 97% after HR (P<.001). Patients with small tumors experienced longer 5-year overall survival rates after HR (72%) as compared with RFA (18%) (P = .006). CONCLUSIONS: The survival rate following HR of solitary colorectal liver metastasis exceeds 70% at 5 years. Radiofrequency ablation for solitary metastasis is associated with a markedly higher LR rate and shorter recurrence-free and overall survival rates compared with HR, even when small lesions (< or = 3 cm) are considered. Every method should be considered to achieve resection of solitary colorectal liver metastasis, including referral to a specialty center, extended hepatectomy, and chemotherapy. PMID- 16702518 TI - Whole body imaging in blunt multisystem trauma patients without obvious signs of injury: results of a prospective study. AB - HYPOTHESIS: The use of liberal whole body imaging (pan scan) in patients based on mechanism is warranted, even in evaluable patients with no obvious signs of chest or abdominal injury. DESIGN: Prospective observational study. SETTING: Academic level I trauma center. PATIENTS: All patients admitted following blunt multisystem trauma. INTERVENTION: Pan scan, including computed tomography (CT) of the head, cervical spine, chest, abdomen, and pelvis, with the following inclusion criteria: (1) no visible evidence of chest or abdominal injury, (2) hemodynamically stable, (3) normal abdominal examination results in a neurologically intact patient or unevaluable abdominal examination results secondary to a depressed level of consciousness, and (4) significant mechanisms of injury. Radiological findings and changes in treatment based on these findings were recorded. MAIN OUTCOME MEASURE: Any alteration in the normal treatment plan as a direct result of CT scan findings. These alterations include early hospital discharge, admission for observation, operative intervention, and additional diagnostic studies or interventions. RESULTS: One thousand patients underwent pan scan during the 18-month observation period, of which 592 were evaluable patients with no obvious signs of abdominal injury. Clinically significant abnormalities were found in 3.5% of head CT scans, 5.1% of cervical spine CT scans, 19.6% of chest CT scans, and 7.1% of abdominal CT scans. Overall treatment was changed in 18.9% of patients based on abnormal CT scan findings. CONCLUSIONS: The use of pan scan based on mechanism in awake, evaluable patients is warranted. Clinically significant abnormalities are not uncommon, resulting in a change in treatment in nearly 19% of patients. PMID- 16702519 TI - Molecular determinants in targeted therapy for esophageal adenocarcinoma. AB - HYPOTHESIS: Cyclooxygenase 2 (COX-2), vascular endothelial growth factor (VEGF), and epidermal growth factor receptor (EGFR) are useful biological determinants in targeted therapy for esophageal adenocarcinoma. DESIGN: Prospective analysis. SETTING: University tertiary referral center. PATIENTS: Sixteen patients with squamous mucosa and normal results of a pH study without mucosal injury (control group), 15 with Barrett esophagus (metaplasia group), and 44 with adenocarcinoma (carcinoma group). INTERVENTIONS: Biopsy specimens were obtained 3 cm above the gastroesophageal junction. Dysplastic tissue was additionally isolated from 9 of the patients in the carcinoma group. After laser-capture microdissection, quantitative real-time polymerase chain reaction was used to measure gene expression across the spectrum of the metaplasia-dysplasia-carcinoma sequence. MAIN OUTCOME MEASURES: Expression of COX-2, VEGF, and EGFR in each patient group. RESULTS: Expression of both COX-2 and VEGF was significantly up-regulated in patients with metaplasia, dysplasia, and cancer compared with controls (P<.01). Expression levels of both were significantly higher in cancer than in the metaplasia group (P<.05) and increased sequentially from metaplasia to dysplasia to cancer. Expression of VEGF was significantly higher in the dysplastic tissue than in nondysplastic Barrett epithelium (P<.05). No change in expression levels of EGFR was seen in the histologic progression to esophageal adenocarcinoma. CONCLUSION: Gene expression data suggest that pharmacologic inhibition of COX-2 and VEGF may be useful adjuncts in targeted therapy for esophageal adenocarcinoma. PMID- 16702520 TI - Which intraoperative parathyroid hormone assay criterion best predicts operative success? A study of 352 consecutive patients. AB - HYPOTHESIS: The 6 published criteria for predicting curative parathyroid resection by means of intraoperative parathyroid hormone (IOPTH) assay are not equivalent. DESIGN: Retrospective review of 352 patients undergoing parathyroidectomy for primary hyperparathyroidism from January 1, 1999, to December 31, 2004. We evaluated 6-month postoperative IOPTH values and serum calcium levels. SETTING: Tertiary referral center. MAIN OUTCOME MEASURES: The IOPTH values at baseline (preincision and preexcision) and at 5 and 10 minutes after parathyroidectomy were reviewed according to the Miami criterion (>50% drop from highest baseline IOPTH level at 10 minutes after excision), criterion 1 (>50% drop from preincision IOPTH level at 10 minutes), criterion 2 (>50% drop from highest baseline IOPTH level at 10 minutes and final IOPTH level within the reference range), criterion 3 (>50% drop from highest baseline IOPTH level at 10 minutes and final IOPTH level less than the preincision value), criterion 4 (>50% drop from highest baseline IOPTH level at 5 minutes), and criterion 5 (>50% drop from preexcision IOPTH level at 10 minutes). RESULTS: Criterion 2 had sensitivity of 88%, specificity of 22%, positive predictive value of 97%, and negative predictive value of 6%. Criterion 2 had good agreement with criteria 1 and 3. Of patients whose IOPTH level drop satisfied criterion 2 but not criterion 1, 14% had postoperative hypercalcemia at 6 months. When criterion 2 was not satisfied but criteria 1, 3, 4, and 5 and the Miami criterion were, failure rates were 0%, 4%, 7%, 6%, and 9%, respectively. CONCLUSIONS: Satisfying criterion 2 had a high operative success but resulted in additional unnecessary surgical exploration. Criterion 1 was better at predicting postoperative normocalcemia than criterion 2. PMID- 16702521 TI - Value of preoperative ultrasonography in the surgical management of initial and reoperative papillary thyroid cancer. AB - BACKGROUND: Cervical recurrences, predominantly in lymph nodes, occur in 14% to 30% of patients with papillary thyroid cancer (PTC). Postoperative surveillance for recurrent PTC increasingly includes thyrotropin-stimulated thyroglobulin and high-resolution ultrasonography (US). This combination commonly can detect recurrent disease as small as 5 mm. HYPOTHESIS: Preoperative US will increase detection and assessment of the extent of lymph node metastasis (LNM) in patients with PTC. DESIGN: Retrospective cohort study. SETTING: Tertiary care academic center. PATIENTS: From January 1, 1999, to December 31, 2004, a total of 770 patients were seen, 551 (381 female and 170 male; median age, 47 years; age range, 9-89 years) who underwent initial surgical management and 219 (154 female and 65 male; median age, 44 years; age range, 5-90 years) who underwent cervical reoperation for PTC. The US images were obtained preoperatively for 486 initial and 216 reoperative patients. Therapeutic radioactive iodine was administered to 151 (68.9%) of the reoperative patients before the subsequent operation (median dose, 5.6 x 10(9) Bq; range, 7.4 x 10(8)-3.7 x 10(10) Bq). RESULTS: Ultrasonography identified nonpalpable lateral jugular LNMs in 70 (14.4%) of the patients undergoing initial exploration. Similarly, in reoperative patients, nonpalpable lateral LNMs were detected via US in 106 (64.2%), and 61 (28.2%) had LNMs detected in the central neck. Even when nodes were palpable preoperatively (37 [6.7%] of the initial and 56 [25.6%] of the reoperative patients), US assessment of the extent of LNM involvement altered the operation in 15 (40.5%) of the initial and 24 (42.9%) of the reoperative patients. The sensitivity, specificity, and positive predictive value for US were 83.5%, 97.7%, and 88.8% in initial patients, and 90.4%, 78.9%, and 93.9% in reoperative patients. CONCLUSIONS: Overall, preoperative US detected nonpalpable LNMs in 231 (32.9%) of the 702 patients with PTC who underwent US, thereby altering the operative procedure performed. In addition, even in patients with palpable LNs, US helped to guide the extent of lymphadenectomy. PMID- 16702522 TI - Primary hyperaldosteronism: effect of adrenal vein sampling on surgical outcome. AB - HYPOTHESIS: Adrenal vein sampling is superior to computed tomography for subtype differentiation of primary hyperaldosteronism. DESIGN: Retrospective review. SETTING: University medical center. PATIENTS: Forty-eight patients (32 men and 16 women) with biochemically confirmed primary hyperaldosteronism. MAIN OUTCOME MEASURES: We compared demographic factors, results of biochemical and imaging studies (computed tomography and adrenal vein sampling), therapy, and patient outcomes. RESULTS: Mean +/- SEM adrenal nodule size was 1.54 +/- 0.2 cm. Adrenal vein sampling was performed in 41 (85%) of 48 patients, and it was successful in 39 (95%) of those 41 patients. Concordance between computed tomography and adrenal vein sampling was observed in 22 (54%) of the 41 patients. Thirty-two patients underwent successful laparoscopic adrenalectomy. There was 1 complication and no deaths. All 32 patients were cured of hypokalemia. CONCLUSION: Adrenal vein sampling is superior to image-based techniques for subtype differentiation of primary hyperaldosteronism. PMID- 16702523 TI - Effects of delaying appendectomy for acute appendicitis for 12 to 24 hours. AB - OBJECTIVE: To determine whether delaying appendectomy for 12 hours to avoid disturbing the operating room schedule and to minimize the number of operations during the night negatively affects the outcome of patients with acute appendicitis. DESIGN: Retrospective study. SETTING: Large teaching community hospital. PATIENTS: The medical records of 380 patients who underwent appendectomies between January 1, 2002, and December 31, 2004, were reviewed. Patients proven to have an inflamed appendix on the pathological report were divided into 2 groups. The early group comprised patients who had undergone appendectomies within 12 hours of presentation to the emergency department, including patients with generalized sepsis. The late group comprised patients who had undergone appendectomies more than 12 to 24 hours after presentation. MAIN OUTCOME MEASURES: Length of stay, operative time, and the rate of perforations and complications. INTERVENTIONS: Laparoscopic or open appendectomies. RESULTS: There were 309 patients included in our study. There were no statistically significant differences between the early and late groups in the length of stay, operative time, the percentage of advanced appendicitis, or the rate of complications. CONCLUSIONS: In selected patients, delaying appendectomies for acute appendicitis for 12 to 24 hours after presentation does not significantly increase the rate of perforations, operative time, or length of stay. It decreases the use of the nursing staff, anesthesia team, and surgical house staff during the night shifts, and it decreases the interruption of the regular operating room schedule. PMID- 16702524 TI - Reverence for life. PMID- 16702525 TI - Image of the month. Subclavian artery aneurysm secondary to cervical rib. PMID- 16702526 TI - Family physicians' training programs are useful for surgeons. PMID- 16702528 TI - New concepts in nasal tip contouring. AB - Control of nasal tip contour has always been a key component of a successful rhinoplasty. Typically, this procedure is performed with an emphasis on narrowing the nasal tip structure. Creating a natural-appearing nasal tip contour is a complex task and requires a 3-dimensional approach. In an effort to identify the characteristics that make an ideal nasal tip, I evaluated numerous aesthetically pleasing nasal tips. After extensive study, I created a series of images to demonstrate how specific contours create highlights and shadows that will help guide the surgeon in creating a natural-appearing nasal tip contour. Many commonly used nasal tip techniques can pinch the tip structures if an overemphasis is placed on narrowing. These changes isolate the dome region of the nasal tip and can create an undesirable shadow between the tip lobule and alar lobule. Prior to contouring the nasal tip, the surgeon must stabilize the base of the nose with a columellar strut, suturing the medial crura to a long caudal septum, caudal extension graft, or an extended columellar strut graft. Stabilizing the nasal base will ensure that tip projection is maintained postoperatively. To contour the nasal tip, dome sutures are frequently used to flatten the lateral crura and eliminate tip bulbosity. Placement of dome sutures can deform the lateral crura and displace the caudal margin of the lateral crura well below the cephalic margin. This can result in a pinched nasal tip with the characteristic demarcation between the tip and the alar lobule. Alar rim grafts can be used to support the alar margin and create a defined ridge that extends from the tip lobule to the alar lobule. This form of restructuring can create a natural-appearing nasal tip contour with a horizontal tip orientation continuing out to the alar lobule. When dome sutures alone are inadequate, lateral crural strut grafts are used to eliminate convexity and prevent deformity of the lateral crura. Shield tip grafts can be used in patients with thick skin and an underprojected nasal tip. Whenever a shield tip graft is used, it must be appropriately camouflaged to avoid undesirable visualization of the graft as the postoperative edema subsides. When contouring the nasal tip, the surgeon should focus more on creating favorable shadows and highlights and less on narrowing. Nasal tips contoured in this manner will look more natural and will better withstand the forces of scar contracture that can negatively affect rhinoplasty outcomes. PMID- 16702529 TI - Limited vs extended face-lift techniques: objective analysis of intraoperative results. AB - OBJECTIVE: To compare the intraoperative outcomes of superficial musculoaponeurotic system plication, imbrication, and deep-plane rhytidectomy techniques. METHODS: Thirty-two patients undergoing primary deep-plane rhytidectomy participated. Each hemiface in all patients was submitted sequentially to 3 progressively more extensive lifts, while other variables were standardized. Four major outcome measures were studied, including the extent of skin redundancy and the repositioning of soft tissues along the malar, mandibular, and cervical vectors of lift. The amount of skin excess was measured without tension from the free edge to a point over the intertragal incisure, along a plane overlying the jawline. Using a soft tissue caliper, repositioning was examined by measurement of preintervention and immediate postintervention distances from dependent points to fixed anthropometric reference points. RESULTS: The mean skin excesses were 10.4, 12.8, and 19.4 mm for the plication, imbrication, and deep-plane lifts, respectively. The greatest absolute soft tissue repositioning was noted along the jawline, with the least in the midface. Analysis revealed significant differences from baseline and between lift types for each of the studied techniques in each of the variables tested. CONCLUSION: These data support the use of the deep-plane rhytidectomy technique to achieve a superior intraoperative lift relative to comparator techniques. PMID- 16702530 TI - Superficial musculoaponeurotic system vs deep-plane face-lift. PMID- 16702531 TI - Deep-plane technique. PMID- 16702532 TI - Reconstructive rhinoplasty: the 3-dimensional nasal tip. AB - OBJECTIVES: To review nasal alar support mechanisms, introduce the concept of tractional forces on the nasal ala, and describe a reconstructive technique to correct nasal tip deformities associated with weakened tractional force on the nasal ala. DESIGN: Photographic study and retrospective medical chart review. RESULTS: We noted that patients with weakened support at the dome of the lower lateral cartilage had lateral alar deformities. Strengthening the cartilaginous deficiency improved the nasal appearance and function in 90% of patients. CONCLUSIONS: Deformities of the nasal tip are among the most difficult to correct. Tractional forces provided by dome strength help to maintain the ala in its normal anatomical position. Structural tip grafts restore the tractional force and, thereby, help to correct the alar deformity. PMID- 16702533 TI - Anterior septal reconstruction: outcomes after a modified extracorporeal septoplasty technique. AB - OBJECTIVE: To describe a modified extracorporeal septoplasty technique and measure its efficacy with a validated quality-of-life instrument. DESIGN: A prospective observational outcomes study of patients with severe septal deviation who subsequently underwent anterior septal reconstruction. Preoperative and postoperative evaluation was performed using photographs and the Nasal Obstruction Symptoms Evaluation scale. RESULTS: Twelve consecutive patients were enrolled. No complications occurred. All patients noted improved airway function postoperatively. There was a significant improvement in mean Nasal Obstruction Symptoms Evaluation score postoperatively (76.6 vs 12.9; P<.01). Examination of postoperative photographs revealed improved midvault and tip anatomy. CONCLUSIONS: The anterior septal reconstruction technique is effective in improving both nasal airway function and aesthetics in patients with severe septonasal deviation. The technique avoids the most common complication of standard extracorporeal septoplasty by preserving the dorsal strut of septal cartilage and its attachment to the nasal bones at the keystone area. PMID- 16702534 TI - Promotion of acellular dermal matrix resolution in vitro by matrix metalloproteinase-2. AB - OBJECTIVE: To determine whether acellular human dermis is degraded by matrix metalloproteinases (MMPs), a large class of matrix-degrading enzymes. METHODS: The degradation of acellular human dermis specimens was evaluated in vitro. Wild type murine fibroblasts with a broad-spectrum MMP inhibitor, GM6001, and MMP-2 deficient fibroblasts were placed on the basement membrane and dermal surfaces of acellular human dermis. Matrix degradation and fibroblast infiltration into the matrix were assessed after a 20-day incubation period. RESULTS: The basement membrane thickness of the specimens cultured with wild-type fibroblasts was significantly less than that of specimens cultured with GM6001 (P<.001), and the infiltration of fibroblasts into the dermal surface was limited by the addition of GM6001 (P=.002). To determine whether MMP-2 was involved in this in vitro phenotype, MMP-2-deficient fibroblasts were assessed in comparison with wild-type fibroblasts. Wild-type fibroblasts degraded the basement membrane surface (P<.001) and infiltrated the dermal surface (P = .003) more efficiently than did MMP-2-deficient fibroblasts. CONCLUSIONS: The results from our in vitro experiments suggest that MMPs and specifically MMP-2 may play an important role in the resorption of acellular human dermis. Addition of MMP inhibitors to implanted dermal matrices may slow fibroblast infiltration and improve their longevity in vivo. PMID- 16702535 TI - Fan septoplasty for correction of the internally and externally deviated nose. PMID- 16702536 TI - Treatment of deep orbital hemangiomas of infancy: an overview. PMID- 16702537 TI - Botulinum toxin to improve results in cleft lip repair. PMID- 16702538 TI - William Sidney Mount's The power of music. PMID- 16702539 TI - The TodS-TodT two-component regulatory system recognizes a wide range of effectors and works with DNA-bending proteins. AB - The TodS and TodT proteins form a previously unrecognized and highly specific two component regulatory system in which the TodS sensor protein contains two input domains, each of which are coupled to a histidine kinase domain. This system regulates the expression of the genes involved in the degradation of toluene, benzene, and ethylbenzene through the toluene dioxygenase pathway. In contrast to the narrow substrate range of this catabolic pathway, the TodS effector profile is broad. TodS has basal autophosphorylation activity in vitro, which is enhanced by the presence of effectors. Toluene binds to TodS with high affinity (Kd = 684 +/- 13 nM) and 1:1 stoichiometry. The analysis of the truncated variants of TodS reveals that toluene binds to the N-terminal input domain (Kd = 2.3 +/- 0.1 microM) but not to the C-terminal half. TodS transphosphorylates TodT, which binds to two highly similar DNA binding sites at base pairs -107 and -85 of the promoter. Integration host factor (IHF) plays a crucial role in the activation process and binds between the upstream TodT boxes and the -10 hexamer region. In an IHF-deficient background, expression from the tod promoter drops 8-fold. In vitro transcription assays confirmed the role determined in vivo for TodS, TodT, and IHF. A functional model is presented in which IHF favors the contact between the TodT activator, bound further upstream, and the alpha-subunit of RNA polymerase bound to the downstream promoter element. Once these contacts are established, the tod operon is efficiently transcribed. PMID- 16702540 TI - The chemokine SDF-1/CXCL12 modulates the firing pattern of vasopressin neurons and counteracts induced vasopressin release through CXCR4. AB - Chemokines play a key role in inflammation. They are expressed not only in neuroinflammatory conditions, but also constitutively by different cell types, including neurons in the normal brain, suggesting that they may act as modulators of neuronal functions. Here, we investigated a possible neuroendocrine role of the chemokine stromal cell-derived factor 1 (SDF-1)/CXCL12. We demonstrated the colocalization of SDF-1 and its receptor CXCR4 with arginine vasopressin (AVP) in the magnocellular neurons of the supraoptic nucleus (SON) and the paraventricular hypothalamic nucleus and on AVP projections to the neurohypophysis. Electrophysiological recordings of SON neurons demonstrated that SDF-1 affects the electrical activity of AVP neurons through CXCR4, resulting in changes in AVP release. We observed that SDF-1 can blunt the autoregulation of AVP release in vitro and counteract angiotensin II-induced plasma AVP release in vivo. Furthermore, a short-term physiological increase in AVP release induced by enhanced plasma osmolarity, which was produced by the administration of 1 M NaCl i.p., was similarly blocked by central injection of SDF-1 through CXCR4. A change in water balance by long-term salt loading induced a decrease in both SDF-1 and CXCR4 parallel to that of AVP immunostaining in SON. From these data, we demonstrate that chemokine actions in the brain are not restricted to inflammatory processes. We propose to add to the known autoregulation of AVP on its own neurons, a second autocrine system induced by SDF-1 able to modulate central AVP neuronal activity and release. PMID- 16702541 TI - Genetic variants of Tgfb1 act as context-dependent modifiers of mouse skin tumor susceptibility. AB - The human TGFB1 gene is polymorphic, and genetic variants are associated with altered cancer risk. However, human genetic association studies have had variable outcomes because TGFbeta1 action is context-dependent. We used the murine skin model of chemical carcinogenesis in genetic linkage analysis of three independent Mus musculus NIH/Ola x (Mus spretus x M. musculus NIH/Ola)F1 backcrosses, to identify a skin tumor susceptibility locus, Skts14, on proximal chromosome 7. Tgfb1 maps at the peak of linkage. The mouse Tgfb1 gene is polymorphic, resulting in cis-regulated differential allelic mRNA expression between M. spretus and M. musculus in F1 mouse skin. This phenomenon is reflected in differential phospho SMAD2 levels, downstream of TGFbeta signaling, between these two mouse species. In normal F1 mouse skin, the Tgfb1SPR allele is expressed at higher levels than the Tgfb1NIH allele, and this differential is accentuated by phorbol 12-myristate 13-acetate treatment. In benign F1 papillomas, this imbalance is reversed, possibly by selection against expression of a hyperactive Tgfb1SPR allele in TGFbeta growth-responsive tumors. We demonstrate that skin tumor susceptibility is altered by Tgfb1 gene dosage, but that manifestation of Tgfb1-linked skin tumor susceptibility in M. musculus NIH/Ola x (M. spretus x M. musculus NIH/Ola)F1 backcross mice depends on interactions with another unlinked tumor modifying locus, Skts15, that overlaps Tgfbm3 on chromosome 12. These findings illustrate the power of complex genetic interactions in determining disease outcome and have major implications to the assessment of disease risk in individuals harboring variant TGFB1 alleles. PMID- 16702543 TI - Velocity of vortices in inhomogeneous Bose-Einstein condensates. AB - We derive, from the Gross-Pitaevskii equation, an exact expression for the velocity of any vortex in a Bose-Einstein condensate, in equilibrium or not, in terms of the condensate wave function at the center of the vortex. In general, the vortex velocity is a sum of the local superfluid velocity, plus a correction related to the density gradient near the vortex. A consequence is that in rapidly rotating, harmonically trapped Bose-Einstein condensates, unlike in the usual situation in slowly rotating condensates and in hydrodynamics, vortices do not move with the local fluid velocity. We indicate how Kelvin's conservation of circulation theorem is compatible with the velocity of the vortex center being different from the local fluid velocity. Finally, we derive an exact wave function for a single vortex near the rotation axis in a weakly interacting system, from which we derive the vortex precession rate. PMID- 16702542 TI - High-resolution video monitoring of hematopoietic stem cells cultured in single cell arrays identifies new features of self-renewal. AB - To search for new indicators of self-renewing hematopoietic stem cells (HSCs), highly purified populations were isolated from adult mouse marrow, micromanipulated into a specially designed microscopic array, and cultured for 4 days in 300 ng/ml Steel factor, 20 ng/ml IL-11, and 1 ng/ml flt3-ligand. During this period, each cell and its progeny were imaged at 3-min intervals by using digital time-lapse photography. Individual clones were then harvested and assayed for HSCs in mice by using a 4-month multilineage repopulation endpoint (>1% contribution to lymphoid and myeloid lineages). In a first experiment, 6 of 14 initial cells (43%) and 17 of 61 clones (28%) had HSC activity, demonstrating that HSC self-renewal divisions had occurred in vitro. Characteristics associated with HSC activity included longer cell-cycle times and the absence of uropodia on a majority of cells within the clone during the final 12 h of culture. Combining these criteria maximized the distinction of clones with HSC activity from those without and identified a subset of 27 of the 61 clones. These 27 clones included all 17 clones that had HSC activity; a detection efficiency of 63% (2.26 times more frequently than in the original group). The utility of these characteristics for discriminating HSC-containing clones was confirmed in two independent experiments where all HSC-containing clones were identified at a similar 2- to 3 fold-greater efficiency. These studies illustrate the potential of this monitoring system to detect new features of proliferating HSCs that are predictive of self-renewal divisions. PMID- 16702544 TI - Factor B of the alternative complement pathway regulates development of airway hyperresponsiveness and inflammation. AB - Exposure to inhaled allergens leads to increases in airway hyperresponsiveness (AHR) and inflammation, associated with increased levels of biologically active fragments derived from the complement C3 and C5 family of proteins. Further, complement activation during allergen challenge in sensitized animals is necessary for the development of AHR and airway inflammation. To define the complement pathway involved, we studied mice deficient in complement factor 4 (C4 /-), a critical component of the classical pathway, or factor B (fB-/-), an essential protein in the alternative complement pathway. WT, C4-/-, and fB-/- mice were sensitized to ovalbumin and subsequently exposed to nebulized ovalbumin (1% in saline) on 3 consecutive days. After allergen sensitization and challenge, fB-/- mice demonstrated significantly lower airway responsiveness to methacholine and less airway inflammation. In contrast, C4-/- mice showed no reduction in AHR and airway inflammation compared with WT mice. Tissue inflammation, goblet cell hyperplasia, and IL-4, IL-5, and IL-13 levels in BAL fluid were significantly reduced in fB-/- mice compared with C4-/- and WT mice. The development of AHR and airway inflammation in sensitized fB-/- mice could be restored after intranasal administration of purified factor B before the airway challenge. In addition, administration of a neutralizing anti-factor B mAb to sensitized mice before airway challenge reduced the development of AHR and airway inflammation. These results demonstrate that in sensitized hosts complement activation through the alternative pathway after allergen exposure is critical to the development of AHR and airway inflammation. PMID- 16702545 TI - Hotspots for copy number variation in chimpanzees and humans. AB - Copy number variation is surprisingly common among humans and can be involved in phenotypic diversity and variable susceptibility to complex diseases, but little is known of the extent of copy number variation in nonhuman primates. We have used two array-based comparative genomic hybridization platforms to identify a total of 355 copy number variants (CNVs) in the genomes of 20 wild-born chimpanzees (Pan troglodytes) and have compared the identified chimpanzee CNVs to known human CNVs from previous studies. Many CNVs were observed in the corresponding regions in both chimpanzees and humans; especially those CNVs of higher frequency. Strikingly, these loci are enriched 20-fold for ancestral segmental duplications, which may facilitate CNV formation through nonallelic homologous recombination mechanisms. Therefore, some of these regions may be unstable "hotspots" for the genesis of copy number variation, with recurrent duplications and deletions occurring across and within species. PMID- 16702546 TI - Fluoxetine targets early progenitor cells in the adult brain. AB - Chronic treatment with antidepressants increases neurogenesis in the adult hippocampus. This increase in the production of new neurons may be required for the behavioral effects of antidepressants. However, it is not known which class of cells within the neuronal differentiation cascade is targeted by the drugs. We have generated a reporter mouse line, which allows identification and classification of early neuronal progenitors. It also allows accurate quantitation of changes induced by neurogenic agents in these distinct subclasses of neuronal precursors. We use this line to demonstrate that the selective serotonin reuptake inhibitor antidepressant fluoxetine does not affect division of stem-like cells in the dentate gyrus but increases symmetric divisions of an early progenitor cell class. We further demonstrate that these cells are the sole class of neuronal progenitors targeted by fluoxetine in the adult brain and suggest that the fluoxetine-induced increase in new neurons arises as a result of the expansion of this cell class. This finding defines a cellular target for antidepressant drug therapies. PMID- 16702547 TI - The positioning of cytoplasmic protein clusters in bacteria. AB - Cell division is a carefully orchestrated procedure. Bacterial cells have intricate mechanisms to ensure that genetic material is copied, proofread, and accurately partitioned into daughter cells. Partitioning now appears to also occur for some cytoplasmic proteins. Previously, using chromosomal fluorescent protein fusions, we demonstrated that a subset of Rhodobacter sphaeroides chemotaxis proteins colocalize to a discrete region within the bacterial cytoplasm. Using TlpT-yellow fluorescent protein as a marker for the position of the cytoplasmic protein clusters, we show most cells contain either one cluster localized at mid-cell or two clusters at the one-fourth and three-fourths positions of cell length. The number and positioning of these protein clusters depend on a previously unrecognized bacterial protein positioning factor, PpfA, which has homology to bacterial type I DNA partitioning factors. These data suggest that there is a mechanism involved in partitioning some cytoplasmic proteins upon cell division that is analogous to a mechanism seen for plasmid and chromosomal DNA. PMID- 16702548 TI - Failing to deactivate: resting functional abnormalities in autism. AB - Several regions of the brain (including medial prefrontal cortex, rostral anterior cingulate, posterior cingulate, and precuneus) are known to have high metabolic activity during rest, which is suppressed during cognitively demanding tasks. With functional magnetic resonance imaging (fMRI), this suppression of activity is observed as "deactivations," which are thought to be indicative of an interruption of the mental activity that persists during rest. Thus, measuring deactivation provides a means by which rest-associated functional activity can be quantitatively examined. Applying this approach to autism, we found that the autism group failed to demonstrate this deactivation effect. Furthermore, there was a strong correlation between a clinical measure of social impairment and functional activity within the ventral medial prefrontal cortex. We speculate that the lack of deactivation in the autism group is indicative of abnormal internally directed processes at rest, which may be an important contribution to the social and emotional deficits of autism. PMID- 16702549 TI - Direct and fast detection of neuronal activation in the human brain with diffusion MRI. AB - Using MRI, we found that a slowly diffusing water pool was expanding (1.7 +/- 0.3%) upon activation on the human visual cortex at the detriment of a faster diffusing pool. The time course of this water phase transition preceded the activation-triggered vascular response detected by usual functional MRI by several seconds. The observed changes in water diffusion likely reflect early biophysical events that take place in the activated cells, such as cell swelling and membrane expansion. Although the exact mechanisms remain to clarify, access to such an early and direct physiological marker of cortical activation with MRI will provide opportunities for functional neuroimaging of the human brain. PMID- 16702550 TI - Evidence that the Y chromosome influences autoimmune disease in male and female mice. AB - Experimental allergic encephalomyelitis (EAE), an autoimmune model of multiple sclerosis, is a complex disease influenced by genetic, intrinsic, and environmental factors. In this study, we questioned whether parent-of-origin effects influence EAE, using reciprocal F2 intercross progeny generated between EAE-susceptible SJL/J (S) and EAE-resistant B10.S/SgMcdJ (B) mice. EAE susceptibility and severity were found to be different in female BS x BS intercross mice as compared with females from the three other birth crosses (BS x SB, SB x SB, and SB x BS), and in fact, both traits in female mice resembled those of their male siblings. This masculinization is associated with transmission of the SJL/J Y chromosome and an increased male-to-female sex ratio. Related studies using progeny of C57BL/6J Y-chromosome substitution strains demonstrate that the Y chromosome again influences EAE in both male and female mice, and that the disease course in females resembles that of their male littermates. Importantly, these data provide experimental evidence supporting the existence of a Y-chromosome polymorphism capable of modifying autoimmune disease susceptibility in both males and females. PMID- 16702551 TI - X-ray structure of a native calicivirus: structural insights into antigenic diversity and host specificity. AB - Caliciviruses, grouped into four genera, are important human and veterinary pathogens with a potential for zoonosis. In these viruses, capsid-related functions such as assembly, antigenicity, and receptor interactions are predominantly encoded in a single protein that forms an icosahedral capsid. Understanding of the immunologic functions and pathogenesis of human caliciviruses in the Norovirus and Sapovirus genera is hampered by the lack of a cell culture system or animal models. Much of our understanding of these viruses, including the structure, has depended on recombinant capsids. Here we report the atomic structure of a native calicivirus from the Vesivirus genus that exhibits a broad host range possibly including humans and map immunological function onto a calicivirus structure. The vesivirus structure, despite a similar architectural design as seen in the recombinant norovirus capsid, exhibits novel features and indicates how the unique modular organization of the capsid protein with interdomain flexibility, similar to an antibody structure with a hinge and an elbow, integrates capsid-related functions and facilitates strain diversity in caliciviruses. The internally located N-terminal arm participates in a novel network of interactions through domain swapping to assist the assembly of the shell domain into an icosahedral scaffold, from which the protruding domain emanates. Neutralization epitopes localize to three hypervariable loops in the distal portion of the protruding domain surrounding a region that exhibits host specific conservation. These observations suggest a mechanism for antigenic diversity and host specificity in caliciviruses and provide a structural framework for vaccine development. PMID- 16702552 TI - Bayesian error analysis model for reconstructing transcriptional regulatory networks. AB - Transcription regulation is a fundamental biological process, and extensive efforts have been made to dissect its mechanisms through direct biological experiments and regulation modeling based on physical-chemical principles and mathematical formulations. Despite these efforts, transcription regulation is yet not well understood because of its complexity and limitations in biological experiments. Recent advances in high throughput technologies have provided substantial amounts and diverse types of genomic data that reveal valuable information on transcription regulation, including DNA sequence data, protein-DNA binding data, microarray gene expression data, and others. In this article, we propose a Bayesian error analysis model to integrate protein-DNA binding data and gene expression data to reconstruct transcriptional regulatory networks. There are two unique aspects to this proposed model. First, transcription is modeled as a set of biochemical reactions, and a linear system model with clear biological interpretation is developed. Second, measurement errors in both protein-DNA binding data and gene expression data are explicitly considered in a Bayesian hierarchical model framework. Model parameters are inferred through Markov chain Monte Carlo. The usefulness of this approach is demonstrated through its application to infer transcriptional regulatory networks in the yeast cell cycle. PMID- 16702553 TI - Salt screening and specific ion adsorption determine neutral-lipid membrane interactions. AB - The simplest, single-component biological membrane challenges accepted models of macromolecular interactions: lipid lamellar phases swell when immersed in monovalent salt solutions. Moreover, typical of a Hofmeister series, Br salts swell multilayers more than Cl salts, offering an excellent opportunity to investigate long-standing questions of ionic specificity. In accord with earlier measurements of liposome mobilities in electric fields, we find an added electrostatic repulsion of membranes due to anion binding, with a much stronger Br binding compared with Cl. However, contrary to the expectation that electrostatic repulsion should vanish in high salinity, swelling of lipid multilayers is monotonic with increasing salt concentration for both Br and Cl salts. The apparent contradiction is resolved by recognizing that although the electrostatic repulsion is progressively screened by increasing salt concentration, so is the van der Waals (vdW) attraction. Negligible in low salt, weakening of vdW forces becomes significant by the time electrostatic forces vanish. The result is a smooth monotonic swelling curve with no apparent distinction between low and high salt concentration regimes. Furthermore, when compared with theoretical predictions, measured vdW forces decay much too slowly with added salt. However, by accounting for the recently measured salt deficit near lipid bilayers, the expected scaling with Debye screening length is recovered. The combination of ion-specific binding and nonspecific ionic screening of low-frequency fluctuations explains salt effects on lipid membrane interactions and, by extension, explains specific (Hofmeister) effects at macromolecular interfaces between low and high dielectric. PMID- 16702554 TI - Binding crossmodal object features in perirhinal cortex. AB - Knowledge of objects in the world is stored in our brains as rich, multimodal representations. Because the neural pathways that process this diverse sensory information are largely anatomically distinct, a fundamental challenge to cognitive neuroscience is to explain how the brain binds the different sensory features that comprise an object to form meaningful, multimodal object representations. Studies with nonhuman primates suggest that a structure at the culmination of the object recognition system (the perirhinal cortex) performs this critical function. In contrast, human neuroimaging studies implicate the posterior superior temporal sulcus (pSTS). The results of the functional MRI study reported here resolve this apparent discrepancy by demonstrating that both pSTS and the perirhinal cortex contribute to crossmodal binding in humans, but in different ways. Significantly, only perirhinal cortex activity is modulated by meaning variables (e.g., semantic congruency and semantic category), suggesting that these two regions play complementary functional roles, with pSTS acting as a presemantic, heteromodal region for crossmodal perceptual features, and perirhinal cortex integrating these features into higher-level conceptual representations. This interpretation is supported by the results of our behavioral study: Patients with lesions, including the perirhinal cortex, but not patients with damage restricted to frontal cortex, were impaired on the same crossmodal integration task, and their performance was significantly influenced by the same semantic factors, mirroring the functional MRI findings. These results integrate nonhuman and human primate research by providing converging evidence that human perirhinal cortex is also critically involved in processing meaningful aspects of multimodal object representations. PMID- 16702555 TI - Complex genetic networks underlying the defensive system of rice (Oryza sativa L.) to Xanthomonas oryzae pv. oryzae. AB - Complete resistance (CR) and partial resistance (PR) of rice (Oryza sativa L.) to its bacterial pathogen, Xanthomonas oryzae pv. oryzae (Xoo), was genetically dissected by using 2 mapping populations and 10 Xoo races. Two CR genes, 50 quantitative resistance loci, and 60 digenic interactions were identified, which showed various degrees of race specificity to the Xoo races. The complex epistasis between these loci led us to the discovery of complex genetic networks underlying the rice defensive system to Xoo. The networks consisted of two major components: one representing interactions between alleles at the R loci of rice and alleles at the corresponding avirulence loci of Xoo for CR and the other comprising interactions between quantitative resistance loci in rice and their corresponding aggressiveness loci in Xoo for PR. The race specificity of PR and its strong genetic overlap with CR indicate that PR is essentially "weaker" CR. The genetic networks discovered are expected to maintain a high level of the allelic diversity at avirulent loci in the pathogen by stabilizing selection, which may maintain a high allelic diversity at R loci in the host by the frequency-dependent selection. PMID- 16702556 TI - Muscle representation in the macaque motor cortex: an anatomical perspective. AB - How are the neurons that directly influence the motoneurons of a muscle distributed in the primary motor cortex (M1)? To answer this classical question we used retrograde transneuronal transport of rabies virus from single muscles of macaques. This enabled us to define cortico-motoneuronal (CM) cells that make monosynaptic connections with the motoneurons of the injected muscle. We examined the distribution of CM cells that project to motoneurons of three thumb and finger muscles. We found that the CM cells for these digit muscles are restricted to the caudal portion of M1, which is buried in the central sulcus. Within this region of M1, CM cells for one muscle display a remarkably widespread distribution and fill the entire mediolateral extent of the arm area. In fact, CM cells for digit muscles are found in regions of M1 that are known to contain the shoulder representation. The cortical territories occupied by CM cells for different muscles overlap extensively. Thus, we found no evidence for a focal representation of single muscles in M1. Instead, the overlap and intermingling among the different populations of CM cells may be the neural substrate to create a wide variety of muscle synergies. We found two additional surprising results. First, 15-16% of the CM cells originate from area 3a, a region of primary somatosensory cortex. Second, the size range of CM cells includes both "fast" and "slow" pyramidal tract neurons. These observations are likely to lead to dramatic changes in views about the function of the CM system. PMID- 16702557 TI - The negative regulator of plant cold responses, HOS1, is a RING E3 ligase that mediates the ubiquitination and degradation of ICE1. AB - Plant responses to cold stress are mediated by a transcriptional cascade, in which the transcription factor ICE1 and possibly related proteins activate the expression of C-repeat (CRT)-binding factors (CBFs), leading to the transcription of downstream effector genes. The variant RING finger protein high expression of osmotically responsive gene (HOS)1 was identified genetically as a negative regulator of cold responses. We present evidence here that HOS1 is an E3 ligase required for the ubiquitination of ICE1. HOS1 physically interacts with ICE1 and mediates the ubiquitination of ICE1 both in vitro and in vivo. We found that cold induces the degradation of ICE1 in plants, and this degradation requires HOS1. Consistent with enhanced cold-responsive gene expression in loss-of-function hos1 mutant plants, overexpression of HOS1 represses the expression of CBFs and their downstream genes and confers increased sensitivity to freezing stress. Our results indicate that cold stress responses in Arabidopsis are attenuated by a ubiquitination/proteasome pathway in which HOS1 mediates the degradation of the ICE1 protein. PMID- 16702558 TI - A single sodium channel mutation produces hyper- or hypoexcitability in different types of neurons. AB - Disease-producing mutations of ion channels are usually characterized as producing hyperexcitability or hypoexcitability. We show here that a single mutation can produce hyperexcitability in one neuronal cell type and hypoexcitability in another neuronal cell type. We studied the functional effects of a mutation of sodium channel Nav1.7 associated with a neuropathic pain syndrome, erythermalgia, within sensory and sympathetic ganglion neurons, two cell types where Nav1.7 is normally expressed. Although this mutation depolarizes resting membrane potential in both types of neurons, it renders sensory neurons hyperexcitable and sympathetic neurons hypoexcitable. The selective presence, in sensory but not sympathetic neurons, of the Nav1.8 channel, which remains available for activation at depolarized membrane potentials, is a major determinant of these opposing effects. These results provide a molecular basis for the sympathetic dysfunction that has been observed in erythermalgia. Moreover, these findings show that a single ion channel mutation can produce opposing phenotypes (hyperexcitability or hypoexcitability) in the different cell types in which the channel is expressed. PMID- 16702560 TI - Serial coalescent simulations suggest a weak genealogical relationship between Etruscans and modern Tuscans. AB - The Etruscans, the only preclassical European population that has been genetically characterized so far, share only two haplotypes with their modern geographic counterparts, the Tuscans, who, nonetheless, appear to be their closest relatives. We modeled 10 demographic scenarios spanning the last 2,500 years and tested by serial coalescent simulation whether any are consistent with the patterns of genetic diversity observed within and between the Etruscan and the modern Tuscan populations. Models in which the Etruscans are the direct ancestors of modern Tuscans appear compatible with the observed data only when they also include a very high mutation rate and an ancient founder effect. A better fit was obtained when the ancient and the modern samples were extracted from two independently evolving populations, connected by little migration. Simulated and observed parameters were also similar for a scenario in which the ancient samples came from a subset, e.g., a social elite, genetically differentiated from the bulk of the Etruscan population. In principle, these results may be biased by factors such as gross and systematic errors in the ancient DNA sequences and failure to sample suitable modern individuals. If neither proves to be the case, this study strongly suggests that either the mitochondrial mutation rate is much higher than currently believed or the Etruscans left very few modern mitochondrial descendants. PMID- 16702559 TI - Complex genomic alterations and gene expression in acute lymphoblastic leukemia with intrachromosomal amplification of chromosome 21. AB - We have previously identified a unique subtype of acute lymphoblastic leukemia (ALL) associated with a poor outcome and characterized by intrachromosomal amplification of chromosome 21 including the RUNX1 gene (iAMP21). In this study, array-based comparative genomic hybridization (aCGH) (n = 10) detected a common region of amplification (CRA) between 33.192 and 39.796 Mb and a common region of deletion (CRD) between 43.7 and 47 Mb in 100% and 70% of iAMP21 patients, respectively. High-resolution genotypic analysis (n = 3) identified allelic imbalances in the CRA. Supervised gene expression analysis showed a distinct signature for eight patients with iAMP21, with 10% of overexpressed genes located within the CRA. The mean expression of these genes was significantly higher in iAMP21 when compared to other ALL samples (n = 45). Although genomic copy number correlated with overall gene expression levels within areas of loss or gain, there was considerable individual variation. A unique subset of differentially expressed genes, outside the CRA and CRD, were identified when gene expression signatures of iAMP21 were compared to ALL samples with ETV6-RUNX1 fusion (n = 21) or high hyperdiploidy with additional chromosomes 21 (n = 23). From this analysis, LGMN was shown to be overexpressed in patients with iAMP21 (P = 0.0012). Genomic and expression data has further characterized this ALL subtype, demonstrating high levels of 21q instability in these patients leading to proposals for mechanisms underlying this clinical phenotype and plausible alternative treatments. PMID- 16702561 TI - Adenotonsillectomy for obstructive sleep apnea syndrome in young children: prevalence of pulmonary complications. AB - OBJECTIVE: To determine, in a series of children younger than 6 years undergoing adenotonsillectomy for treatment of clinical obstructive sleep apnea syndrome (OSAS), the effect of age on prevalence of postoperative respiratory complications. The primary objective was to define a practice standard for postoperative hospital admission. DESIGN: Retrospective analysis. SETTING: Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio. PATIENTS: All children younger than 6 years who underwent adenotonsillectomy to treat OSAS from June 1, 1999, to May 31, 2001. MAIN OUTCOME MEASURES: The percentage of children younger than 3 years undergoing adenotonsillectomy to treat OSAS who experience a postoperative respiratory complication. RESULTS: Of 2315 patients younger than 6 years undergoing an adenotonsillectomy for treatment of OSAS, 149 (6.4%) developed a postoperative respiratory complication. Even though there was a lower incidence of comorbid medical conditions in this cohort, children younger than 3 years were at a greater risk for developing a postoperative respiratory complication compared with those aged 3 to 5 years (9.8% vs 4.9%, P<.001). Logistic regression analysis revealed that children younger than 3 years had a nearly 2-fold increased risk for respiratory complications postoperatively (odds ratio, 1.98; 95% confidence interval, 1.41-2.77) when controlling for race and sex. CONCLUSIONS: Adenotonsillectomy to treat OSAS is associated with a significantly higher rate of postoperative respiratory complication in children younger than 3 years compared with children aged 3 to 5 years. Our results support hospital admission for all patients younger than 3 years undergoing adenotonsillectomy for treatment of OSAS. PMID- 16702562 TI - Mannose-binding lectin and upper respiratory tract infections in children and adolescents: a review. AB - OBJECTIVE: To review the literature on mannose-binding lectin (MBL) polymorphisms and susceptibility for upper respiratory tract infection (URI) in children and adolescents. DATA SOURCES: We searched PubMed from 1966 and EMBASE from 1974 to July 2005, using the terms respiratory tract infection, respiratory infection, upper respiratory infection, MBL, and mannose-binding lectin. STUDY SELECTION: Initially, 110 studies were identified. Two reviewers independently screened identified titles and abstracts. Potentially relevant studies were obtained and the full text examined. Inclusion criteria were human subjects, 18 years or younger, URI, and MBL polymorphisms. Seven of the initially identified studies met the inclusion criteria. DATA EXTRACTION: Information was gathered for each study on study design, population, possible confounders, and outcomes measured. DATA SYNTHESIS: Because there was significant heterogeneity between the identified studies, we had to describe the identified studies separately. The largest case-control studies (n = 3) as well as the cohort study (n = 1) suggest an association between MBL polymorphisms and URI, especially in young children. Results of the smaller studies (n = 3) are inconsistent. CONCLUSIONS: The association between MBL polymorphisms and URI in children remains controversial. Large prospective cohort studies with regular documentation of URI and possible confounders such as atopy and environmental factors are required to establish the role of MBL polymorphisms in susceptibility for URI. PMID- 16702563 TI - Evaluation of the efficacy of supraglottoplasty in obstructive sleep apnea syndrome associated with severe laryngomalacia. AB - OBJECTIVE: To evaluate the clinical and polysomnographic evolution of patients with severe laryngomalacia who underwent supraglottoplasty. DESIGN: Retrospective study. SETTING: University hospital. PATIENTS: Seven children with severe laryngomalacia. MAIN OUTCOME MEASURES: The patients were evaluated with a questionnaire given to their parents and with polysomnographic examination before and 3 months after surgery. The clinical data of respiratory and swallowing symptoms as well as the parameters of minimum oxygen saturation and respiratory disturbance index were evaluated after bilateral supraglottoplasty and compared with the preoperative data. RESULTS: Two patients had pharyngolaryngomalacia and required tracheotomy. Four patients had a marked improvement of respiratory and deglutition symptoms. Polysomnographic data showed a significant improvement in the respiratory disturbance index after surgery (P<.05) but not in the minimum oxygen saturation level. However, this improvement was only partially achieved in 3 patients, in whom there were associated airway or neurologic changes. No serious surgical complications were observed. CONCLUSION: Supraglottoplasty led to a marked improvement in all 5 patients without pharyngolaryngomalacia, but the 2 patients with pharyngolaryngomalacia required tracheotomy. PMID- 16702564 TI - Connexin-associated deafness and speech perception outcome of cochlear implantation. AB - OBJECTIVE: To compare performance after cochlear implantation in children with mutations in connexin (Cx) 26 (GJB2) or Cx30 (GJB6) and children with deafness of unknown etiology. DESIGN: Genetic analysis and speech perception evaluation was performed in the children with and without Cx mutations who had undergone cochlear implantation. Speech perception performance was retrospectively analyzed 6, 12, 24, 36, and 48 months after implantation. Test material was selected according to the child's age and cognitive and language abilities. SETTING: The study took place at speech and hearing and genetic centers of a hospital in the central part of Israel and the genetics departments of 3 additional centrally located hospitals. PATIENTS: A total of 30 children who had undergone cochlear implantation were selected for the study, with control patients matched according to age at implantation, duration of implant use, and mode of communication. There was no evidence for additional disabilities or handicaps in either group. MAIN OUTCOME MEASURES: Speech perception measurements included a questionnaire, as well as closed and open-set tests. RESULTS: Overall, the 2 groups showed significant improvement in speech perception results after implantation. Four years after implantation, both groups achieved mean open-set speech perception scores of approximately 60%, 75%, and 90% for monosyllabic, 2 syllables, and words in sentences tests, respectively. CONCLUSIONS: There were no apparent differences in speech perception performance after implantation between the children with Cx mutations and children with deafness of unknown etiology. These data have important implications as a prognostic indicator when counseling candidates for cochlear implantation. PMID- 16702565 TI - Efficacy of postural restriction in treating benign paroxysmal positional vertigo. AB - OBJECTIVE: To investigate the efficacy of postural restriction after canalith repositioning in treating benign paroxysmal positional vertigo (BPPV). DESIGN: Prospective trial of patients with postural restriction vs those without postural restriction after treatment. PATIENTS: Patients with classic BPPV and with BPPV without nystagmus were treated using the modified Epley canalith repositioning procedure. Patients were randomly separated into 2 groups. The first group was instructed to wear a cervical collar and to maintain an upright head position for 2 days. The second group had no motion restriction. After 5 days, the patients were followed up and evaluated using the Dix-Hallpike test. RESULTS: In the first group, 56 of 62 ears healed after the first maneuver, and the remaining ears healed after the second. In the second group, 45 of 57 ears healed after the first maneuver, 6 after the second, and 5 (with subsequent postural restriction) after the third (1 ear did not improve). Five patients in the first group and 3 patients in the second group had BPPV without nystagmus; all of these patients healed after a single maneuver. The difference between the 2 groups in the number of maneuvers required for treatment was statistically significant (P<.05). The number of patients who required a third maneuver was significantly higher in the second group (P<.05). CONCLUSIONS: Postural restriction enhances the therapeutic effect of canalith repositioning in the treatment of posterior semicircular canal BPPV. The long-term efficacy of postural restriction in preventing BPPV recurrence has not been demonstrated. PMID- 16702566 TI - Hearing impairment susceptibility in elderly men and the DFNA18 locus. AB - OBJECTIVE: To identify any chromosomal region that shows evidence for linkage to age-related hearing loss in humans. DESIGN: Evaluation of genetic linkage using sibling-pair methods for hearing loss collected via self-report questionnaire and markers from a genome screening collected from a population-based representative sample of male fraternal twins born from 1917 to 1927. SUBJECTS: Members of a group of 6108 World War II and Korean War veteran twins (2059 complete pairs) who completed a health history questionnaire at a mean age of 74.3 years (range, 69 82 years). A subset of 711 twins (343 complete pairs) later provided a blood sample for DNA extraction in a study of genetic factors in healthy aging. Among the complete pairs were approximately 160 fraternal twins; 50 of these pairs were concordant for age-related hearing loss with at least 1 co-twin reporting bilateral hearing loss and with marker data available for analysis. RESULTS: A region suggestive of linkage was found on chromosome 3q, with a logarithm of the odds score of 2.5 in the same region of this chromosome where the DFNA18 locus resides, which has been reported to cause a form of progressive hereditary hearing loss. CONCLUSIONS: To our knowledge, this is the first sample from the general population that has been used in a genome screening for qualitative hearing loss. The results, if confirmed, suggest that genetic variation in the region of DFNA18 may be responsible for hearing loss with age in the general population. PMID- 16702567 TI - Correlation of numerical aberrations of chromosomes X and 11 and poor prognosis in squamous cell carcinomas of the head and neck. AB - OBJECTIVE: To investigate the prognostic significance of chromosomal aberrations of chromosomes X and 11 in relation to disease-specific survival in head and neck squamous cell carcinoma. SETTING: University hospital. DESIGN: A 10-year retrospective clinical study. Information about clinical findings, treatment, and follow-up has been recorded prospectively. PATIENTS: By means of the fluorescence in situ hybridization technique with centromeric probes for chromosomes X and 11, we analyzed 40 randomly selected patients before treatment for T1 to T4 head and neck squamous cell carcinoma. Numerical aberrations were scored and evaluated in frozen sections. MAIN OUTCOME MEASURES: The significance of prognostic parameters was tested by the log-rank and Kaplan-Meier methods for the univariate analysis. The Cox proportional hazards regression model was used for multivariate analysis. RESULTS: Numerical aberrations of chromosome 11 correlated positively with T and N classification (P = .03 and P = .02, respectively) and with clinical stage (P = .02). Patients with higher frequencies of numerical aberrations for both chromosome X (>48%, mean) and chromosome 11 (>57%, mean) had shortened disease specific survival compared with those with lower frequencies of numerical aberrations (P = .008 and P<.001, respectively). Of patients who died from disease within 3 years, 7 (50%) had a trisomic value of chromosome 11 of 35% or higher of nuclei (P<.001). Moreover, patients with a higher value (>or=8%) of amplification of chromosome 11 (>4 signals) were associated with having poor prognosis compared with those with a lower value (P = .02). CONCLUSION: Numerical aberrations of chromosomes X and 11 had prognostic value in head and neck squamous cell carcinoma, and higher frequencies of numerical aberrations correlated with poor prognosis. PMID- 16702568 TI - Solitary fibrous tumors of the head and neck: a clinicopathologic and radiologic review. AB - OBJECTIVE: To describe the clinicopathologic and radiologic features of solitary fibrous tumors of the head and neck. DESIGN: Retrospective analysis. SETTING: Tertiary referral center that performs head and neck surgical oncology. PATIENTS: Twelve patients with solitary fibrous tumors of the head and neck identified from the pathology and soft tissue tumor databases at Memorial Sloan-Kettering Cancer Center, New York, NY, from 1990 to 2004. All cases were reviewed by 3 experienced pathologists, 1 of whom is an experienced soft tissue tumor pathologist. The diagnosis was confirmed by microscopic features on hematoxylin-eosin staining and by positive staining for CD34 and Bcl2 on immunohistochemical analysis. Tumors were scored for mitotic activity, cellularity, nuclear pleomorphism, necrosis, and the presence of a malignant component. Details on patient characteristics, tumor characteristics, previous treatment and surgery, adjuvant treatment, and outcome were recorded from clinical records. RESULTS: Solitary fibrous tumors occurred in patients over a wide age range (27-78 years; median age, 52 years). Seven patients (58%) were women, and 5 (42%) were men. Most tumors presented as a slow-growing painless mass with a duration ranging from 2 months to 5 years. The tumors ranged from 1 x 1 cm to 6 x 5 cm. Patients presented with a subcutaneous mass of the scalp or face in 4 cases, intraoral mass in 4, sinonasal mass in 3, and paraspinal mass in 1. Computed tomographic and/or magnetic resonance imaging scans of 7 of the 12 patients showed well-circumscribed tumors that enhanced strongly with contrast. Treatment for all of the patients was surgical resection. Pathologic findings showed that 9 tumors were benign and 3 were malignant. Three patients had a positive surgical resection margin. All patients were alive at a median follow-up of 8 months (range, 1-76 months). Local recurrence occurred in 1 patient who had positive surgical margins 3 years after the initial surgery. CONCLUSIONS: Solitary fibrous tumors of the head and neck region are rare and most commonly benign. The diagnosis depends on microscopic and immunohistochemical features, although imaging may help. Patients with these tumors can be safely treated with local excision, but tumors with positive margins require close follow-up over several years owing to the potential for late local recurrence. PMID- 16702569 TI - Examining the need for neck dissection in the era of chemoradiation therapy for advanced head and neck cancer. AB - OBJECTIVES: To (1) determine clinical factors that predict pathologic complete response (pCR) on neck dissection after sequential chemoradiotherapy (SCRT) for advanced head and neck cancer and (2) compare survival parameters between those who underwent neck dissection and those who did not among those patients with a clinical complete response (cCR) in the neck after SCRT, thus assessing the benefit of neck dissection in patients with a cCR in the neck. DESIGN: Retrospective review with a mean follow-up of 3.5 years. SETTING: Regional cancer center. PATIENTS: The study population comprised 55 patients undergoing SCRT for advanced head and neck cancer with N2 or N3 neck disease. Three patients developed progressive disease and were excluded, and 28 patients underwent neck dissection. INTERVENTIONS: Patients were assessed by physical examination and radiographically after SCRT. MAIN OUTCOME MEASURES: Physical examination and radiographic assessments of residual neck disease were compared with pathologic findings in those patients who underwent neck dissection. Survival comparisons were made between patients with a cCR in the neck who underwent neck dissection and those who did not. RESULTS: Of 28 patients who underwent neck dissection, 8 had persistent pathologically positive nodal disease: 5 (45%) of 11 had N3 and 3 (18%) of 17 had N2 disease. Individual clinical neck assessments after SCRT were fairly predictive of a negative pathologic finding at neck dissection. The negative predictive values were physical examination (75%), computed tomography or magnetic resonance imaging (71%), and positron emission tomography (75%). However, when physical examination, imaging studies, and positron emission tomography all indicated a complete response, this accurately predicted a pCR on neck dissection. There appeared to be no improvement in survival parameters when a neck dissection was performed on patients with a cCR in the neck. CONCLUSIONS: Patients with N3 disease are at high risk for residual neck metastasis after SCRT. Patients with N2 disease can be assessed with physical examination, imaging studies, and positron emission tomography. If these all indicate a cCR, then neck dissection is likely not needed. Neck dissection did not appear to further improve survival parameters for patients with a cCR in the neck. PMID- 16702570 TI - Characterization of the sniff magnitude test. AB - OBJECTIVE: To evaluate the potential utility of the Sniff Magnitude Test (SMT) as a clinical measure of olfactory function. DESIGN: Between-subject designs were used to compare the SMT and University of Pennsylvania Smell Identification Test (UPSIT) in study participants from a broad range of ages. SUBJECTS: A total of 361 individuals from retirement communities and an urban university and patients from an otolaryngology clinic. INTERVENTION: Study participants completed the SMT and UPSIT using standard procedures. MAIN OUTCOME MEASURES: The UPSIT was scored using standard procedures to calculate the number of correctly identified odors; a score that can range from 0 to 40 correct. The measure of olfactory function generated by the SMT is the "sniff magnitude ratio," defined as the mean sniff magnitude generated by the odor stimuli divided by the mean sniff magnitude to nonodorized air blanks. RESULTS: The SMT generally showed good agreement with UPSIT diagnostic categories, although SMT scores were only modestly elevated in the mild and modest hyposmia range of the UPSIT. Age-related decline in olfactory ability was evident on the UPSIT at younger ages than that seen with the SMT. As predicted, otolaryngology patients with olfactory complaints were found to be impaired on both the UPSIT and SMT. CONCLUSIONS: The SMT provides a novel method for evaluating the sense of smell that shows good general agreement with the UPSIT. Its minimal dependence on language and cognitive abilities provides some advantages over odor identification tests. There is some indication that the UPSIT may be more sensitive to olfactory (and/or nonolfactory) deficits. We conclude that sniffing behavior can be exploited for the clinical evaluation of olfaction. A comparison of performance on odor identification and sniffing tests may provide novel insight into the nature of olfactory problems in a variety of patient populations. PMID- 16702571 TI - Up-regulation of chemokine ligand 20 in chronic rhinosinusitis. AB - OBJECTIVES: To investigate the up-regulation of chemokine ligand 20 (CCL20) in chronic rhinosinusitis mucosa and to localize the distribution of CCL20 in the human paranasal sinus mucosa. DESIGN: Prospective study. SETTING: Tertiary academic institution. PATIENTS: Ten patients who underwent functional endoscopic sinus surgery for chronic rhinosinusitis without nasal polyps and 10 normal control subjects. INTERVENTIONS: Messenger RNA was extracted from the sinus mucosa, and semiquantitative reverse transcriptase-polymerase chain reaction was performed. Immunohistochemical staining was used to localize the CCL20 protein. RESULTS: The expression levels of CCL20 messenger RNA level in chronic rhinosinusitis without nasal polyps were significantly increased compared with those in normal sinus mucosa. The expression of CCL20 protein was greater in chronic rhinosinusitis without nasal polyps mucosa and was localized to the epithelial and submucosal glandular cells. CONCLUSION: CCL20 is an inducible product of human paranasal sinus epithelium that may play a role in modulating mucosal immunity of the sinus mucosa. PMID- 16702572 TI - The effect of acute xerostomia on vocal function. AB - OBJECTIVE: To evaluate whether xerostomia can affect vocal function in an experimental model. DESIGN: Randomized controlled study. SETTING: Academic center. SUBJECTS: Twenty healthy young men. INTERVENTIONS: Glycopyrrolate was administered to induce acute xerostomia in 10 men, and saline was administered to 10 men who served as controls. MAIN OUTCOME MEASURES: Whole salivary flow rate and dry mouth scale were time-serially measured, and subjective and objective vocal functions were checked before and after treatment. The salivary and vocal functions were compared between groups. RESULTS: Significant salivary hypofunction and symptoms developed within 90 minutes after the administration of glycopyrrolate. Vocal effort and phonation threshold pressure significantly increased (P<.005) and voice range profiles decreased in the xerostomia group (P = .003) but not in the control group. Other voice parameters were comparable between groups (P>.05). High correlations were also found between salivary and vocal parameters. CONCLUSION: Our findings suggest that vocal function can be affected by xerostomia. PMID- 16702573 TI - The use of a screening questionnaire to determine the incidence of allergic rhinitis in singers with dysphonia. AB - OBJECTIVES: To report the incidence of allergic rhinitis in singers with nonspecific laryngeal examination findings and to correlate the incidence of allergic rhinitis with their vocal symptoms when present. DESIGN: A retrospective review of all the medical records of singers with nonspecific laryngeal findings who presented to a specialty voice center for either vocal training or therapy between June 2002 and September 2005. SETTING: Specialty voice center. PARTICIPANTS: Forty-five singers with nonspecific laryngeal findings who presented to a specialty voice center for either vocal training or therapy. MAIN OUTCOME MEASURES: A standardized validated questionnaire for evaluation of allergic rhinitis was filled out by all the subjects. A score above 0 was considered positive. RESULTS: The total prevalence of allergic rhinitis was 87% (39 of 45 subjects). The singers with vocal symptoms were approximately 15% more likely to have allergic rhinitis than those with no vocal symptoms (92% vs 84%). Singers with more than 2 vocal symptoms had a 25% higher likelihood of having allergic rhinitis. CONCLUSIONS: The incidence of allergic rhinitis in singers is extremely high. Hidden respiratory allergies may affect the professional voice. Proper awareness and a multidisciplinary approach are indispensable for proper diagnosis and treatment. PMID- 16702574 TI - Tinnitus and cerebellar developmental venous anomaly. PMID- 16702575 TI - Pathology quiz case 1. Epithelial-myoepithelial carcinoma (EMC). PMID- 16702576 TI - Pathology quiz case 2. Kaposi sarcoma (KS) of the pinna and external auditory canal. PMID- 16702578 TI - Surgery of the primary tumor in metastatic breast cancer: closing the barn door after the horse has bolted? PMID- 16702577 TI - Radiology quiz case. Antrochoanal polyp (ACP). PMID- 16702579 TI - The role of prevention in oncology practice: results from a 2004 survey of American Society of Clinical Oncology members. AB - PURPOSE: In 2004, the American Society of Clinical Oncology (ASCO) Cancer Prevention Committee surveyed the members to describe involvement in clinical prevention activities. METHODS: A previously administered survey, with updated items on genetics, chemoprevention, and survivorship, was mailed to a stratified random sample of 2,000 domestic members and a convenience sample of 3,144 international members. RESULTS: A total of 49.7% of domestic members contacted and survey eligible responded (n = 851). Nonresponders were younger (50.5 v 51.7 years; P < .01); 465 international members responded. Overall, 35% had received formal instruction in cancer prevention and control, and most respondents expected increased use of prevention, screening/early detection, and risk reduction/genetic counseling in their practices in the next 5 years. Most reported caring for cancer survivors, including providing general medical care. They also either directly provide or refer patients for cancer prevention and control services (eg, cancer screening, tobacco and nutrition counseling, risk reduction, and chemoprevention). Multivariable modeling found fewer perceived barriers to inclusion of cancer prevention activities in clinical practice among those practicing in an academic setting, seeing a higher proportion of patients without a cancer diagnosis, having formal training in prevention and control, expecting an increase in prevention activities in the next 5 years, and providing community advice on prevention. CONCLUSION: Barriers to the inclusion of cancer prevention and control activities in oncology clinical practice exist. Nevertheless, a substantial proportion of both domestic and international ASCO members report an interest in cancer prevention and control activities, with a desire for more specific educational programs in this emerging area of oncology practice. PMID- 16702580 TI - Complete excision of primary breast tumor improves survival of patients with metastatic breast cancer at diagnosis. AB - PURPOSE: Surgery of the primary tumor usually is not advised for patients with metastatic breast cancer at diagnosis because the disease is considered incurable. In this population-based study, we evaluate the impact of local surgery on survival of patients with metastatic breast cancer at diagnosis. METHODS: We included all 300 metastatic breast cancer patients recorded at the Geneva Cancer Registry between 1977 and 1996. We compared mortality risks from breast cancer between patients who had surgery of the primary breast tumor to those who had not and adjusted these risks for other prognostic factors. RESULTS: Women who had complete excision of the primary breast tumor with negative surgical margins had a 40% reduced risk of death as a result of breast cancer (multiadjusted hazard ratio [HR], 0.6; 95% CI, 0.4 to 1.0) compared with women who did not have surgery (P = .049). This mortality reduction was not significantly different among patients with different sites of metastasis, but in the stratified analysis the effect was particularly evident for women with bone metastasis only (HR, 0.2; 95% CI, 0.1 to 0.4; P = .001). Survival of women who had surgery with positive surgical margins was not different from that of women who did not have surgery. CONCLUSION: Complete surgical excision of the primary tumor improves survival of patients with metastatic breast cancer at diagnosis, particularly among women with only bone metastases. PMID- 16702581 TI - Stroke rates and risk factors in patients treated with radiation therapy for early-stage breast cancer. AB - PURPOSE: To examine whether stroke risk is elevated in American breast cancer patients treated with modern techniques, as well as whether supraclavicular radiation therapy (RT) is associated with increased risk. METHODS: Observed rates of stroke in 820 eligible early-stage breast cancer patients treated at the University of Michigan Hospital (Ann Arbor, MI) were compared with expected rates. Relationships between potential risk factors and actuarial rate of first stroke were analyzed. RESULTS: Median follow-up was 6.8 years. Twenty patients had at least one cerebrovascular accident (CVA) in follow-up; 35 patients had at least one CVA or transient ischemic attack (CVA/TIA). The standardized incidence ratios were 1.74 (0.94 to 2.37) for CVA and 1.68 (1.003 to 2.06) for CVA/TIA. The absolute excess risk per 1,000 patients per year was 1.67 for CVA and 2.76 for CVA/TIA. On bivariate analysis, factors significantly associated with actuarial rate of first CVA included hypertension (P = .002), age (P < .0001), coronary artery disease (P = .001), atrial fibrillation (P = .009), and supraclavicular RT (P = .021). Factors associated with CVA/TIA were hypertension (P < .001), coronary artery disease (P = .002), and age (P < .0001). Tamoxifen use alone was not significant (P = .19), but tamoxifen combined with baseline hypertension led to increased risk of CVA/TIA (log-rank P < .0001). On multivariate analysis, only age (P < .001) and hypertension (P = .003) remained significant predictors of CVA/TIA. Age was the only significant predictor of CVA alone (P < .001). CONCLUSION: American breast cancer survivors may have an elevated risk of stroke compared with the general population, but the absolute excess risk is low. This study found no significant association between supraclavicular RT and stroke after controlling for other factors. PMID- 16702582 TI - Randomized controlled trial of weight training and lymphedema in breast cancer survivors. AB - PURPOSE: Lymphedema is a common condition that breast cancer survivors face. Despite a lack of supporting evidence from prospective observational studies, occupational and leisure time physical activity are feared to be possible risk factors for lymphedema onset or exacerbation. We examined effects of supervised upper- and lower-body weight training on the incidence and symptoms of lymphedema in 45 breast cancer survivors who participated in the Weight Training for Breast Cancer Survivors study. METHODS: Participants were on average 52 years old, 4 to 36 months post-treatment, and had axillary dissection as part of their treatment. Thirteen women had prevalent lymphedema at baseline. The intervention was twice-a week weight training over a period of 6 months. Lymphedema was monitored at baseline and 6 months by measuring the circumference of each arm, and by self report of symptoms and clinical diagnosis. RESULTS: None of the intervention group participants experienced a change in arm circumferences > or = 2.0 cm after a 6-month exercise intervention. Self-reported incidence of a clinical diagnosis of lymphedema or symptom changes over 6 months did not vary by intervention status (P = .40 and P = .22, respectively). CONCLUSION: This is the largest randomized controlled trial to examine associations between exercise and lymphedema in breast cancer survivors. The results of this study support the hypotheses that a 6-month intervention of resistance exercise did not increase the risk for or exacerbate symptoms of lymphedema. These results herald the need to start reevaluating common clinical guidelines that breast cancer survivors avoid upper body resistance activity for fear of increasing risk of lymphedema. PMID- 16702583 TI - Summaries for patients. Hepatitis C virus infection in the United States, 1999 through 2002. PMID- 16702584 TI - Summaries for patients. A simple test to diagnose urinary incontinence in women. PMID- 16702585 TI - Summaries for patients. Long-term outcomes of treating nonacute Chagas disease with benznidazole. PMID- 16702586 TI - The prevalence of hepatitis C virus infection in the United States, 1999 through 2002. AB - BACKGROUND: Defining the primary characteristics of persons infected with hepatitis C virus (HCV) enables physicians to more easily identify persons who are most likely to benefit from testing for the disease. OBJECTIVE: To describe the HCV-infected population in the United States. DESIGN: Nationally representative household survey. SETTING: U.S. civilian, noninstitutionalized population. PARTICIPANTS: 15,079 participants in the National Health and Nutrition Examination Survey between 1999 and 2002. MEASUREMENTS: All participants provided medical histories, and those who were 20 to 59 years of age provided histories of drug use and sexual practices. Participants were tested for antibodies to HCV (anti-HCV) and HCV RNA, and their serum alanine aminotransferase (ALT) levels were measured. RESULTS: The prevalence of anti-HCV in the United States was 1.6% (95% CI, 1.3% to 1.9%), equating to an estimated 4.1 million (CI, 3.4 million to 4.9 million) anti-HCV-positive persons nationwide; 1.3% or 3.2 million (CI, 2.7 million to 3.9 million) persons had chronic HCV infection. Peak prevalence of anti-HCV (4.3%) was observed among persons 40 to 49 years of age. A total of 48.4% of anti-HCV-positive persons between 20 and 59 years of age reported a history of injection drug use, the strongest risk factor for HCV infection. Of all persons reporting such a history, 83.3% had not used injection drugs for at least 1 year before the survey. Other significant risk factors included 20 or more lifetime sex partners and blood transfusion before 1992. Abnormal serum ALT levels were found in 58.7% of HCV RNA positive persons. Three characteristics (abnormal serum ALT level, any history of injection drug use, and history of blood transfusion before 1992) identified 85.1% of HCV RNA-positive participants between 20 and 59 years of age. LIMITATIONS: Incarcerated and homeless persons were not included in the survey. CONCLUSIONS: Many Americans are infected with HCV. Most were born between 1945 and 1964 and can be identified with current screening criteria. History of injection drug use is the strongest risk factor for infection. PMID- 16702587 TI - The sensitivity and specificity of a simple test to distinguish between urge and stress urinary incontinence. AB - BACKGROUND: Urinary incontinence is common in women. Because treatments differ, urge incontinence should be distinguished from stress incontinence. To make this distinction, current guidelines recommend an extensive evaluation that is too time-consuming for primary care practice. OBJECTIVE: To test the accuracy of a simple questionnaire to categorize type of urinary incontinence in women. DESIGN: Multicenter, prospective study of the accuracy of the 3 Incontinence Questions (3IQ) compared with an extended evaluation to distinguish between urge incontinence and stress incontinence. SETTING: 5 academic medical centers in the United States. PARTICIPANTS: 301 women enrolled from April to December 2004 who were older than 40 years of age (mean age, 56 years [SD, 11]) with untreated incontinence for an average of 7 years (SD, 7) and a broad range of incontinence severity. MEASUREMENTS: All participants included in the analyses answered the 3IQ questionnaire, and a urologist or urogynecologist who was blinded to the responses performed the extended evaluation. Sensitivity, specificity, and likelihood ratios were determined for the 3IQ. RESULTS: For classification of urge incontinence and with the extended evaluation as the gold standard, the 3IQ had a sensitivity of 0.75 (95% CI, 0.68 to 0.81), a specificity of 0.77 (CI, 0.69 to 0.84), and a positive likelihood ratio of 3.29 (CI, 2.39 to 4.51). For classification of stress incontinence, the sensitivity was 0.86 (CI, 0.79 to 0.90), the specificity was 0.60 (CI, 0.51 to 0.68), and the positive likelihood ratio was 2.13 (CI, 1.71 to 2.66). LIMITATIONS: Participants were enrolled by urologists and urogynecologists at academic medical centers. CONCLUSIONS: The 3IQ questionnaire is a simple, quick, and noninvasive test with acceptable accuracy for classifying urge and stress incontinence and may be appropriate for use in primary care settings. Similar studies are needed in other populations. We also need a clinical trial comparing the outcomes of treatments based on the 3IQ and the extended evaluation. PMID- 16702588 TI - Long-term cardiac outcomes of treating chronic Chagas disease with benznidazole versus no treatment: a nonrandomized trial. AB - BACKGROUND: Benznidazole is effective for treating acute-stage Chagas disease, but its effectiveness for treating indeterminate and chronic stages remains uncertain. OBJECTIVE: To compare long-term outcomes of patients with nonacute Chagas disease treated with benznidazole versus outcomes of those who did not receive treatment. DESIGN: Clinical trial with unblinded, nonrandom assignment of patients to intervention or control groups. SETTING: Chagas disease center in Buenos Aires, Argentina. PATIENTS: 566 patients 30 to 50 years of age with 3 positive results on serologic tests and without heart failure. MEASUREMENTS: The primary outcome was disease progression, defined as a change to a more advanced Kuschnir group or death. Secondary outcomes included new abnormalities on electrocardiography and serologic reactivity. INTERVENTION: Oral benznidazole, 5 mg/kg of body weight per day for 30 days (283 patients), or no treatment (283 patients). RESULTS: Fewer treated patients had progression of disease (12 of 283 [4%] vs. 40 of 283 [14%]; adjusted hazard ratio, 0.24 [95% CI, 0.10 to 0.59]; P = 0.002) or developed abnormalities on electrocardiography (15 of 283 [5%] vs. 45 of 283 [16%]; adjusted hazard ratio, 0.27 [CI, 0.13 to 0.57]; P = 0.001) compared with untreated patients. Left ventricular ejection fraction (hazard ratio, 0.97 [CI, 0.94 to 0.99]; P < 0.002) and left ventricular diastolic diameter (hazard ratio, 2.45 [CI, 1.53 to 3.95]; P < 0.001) were also associated with disease progression. Conversion to negative results on serologic testing was more frequent in treated patients than in untreated patients (32 of 218 [15%] vs. 12 of 212 [6%]; adjusted hazard ratio, 2.1 [CI, 1.06 to 4.06]; P = 0.034). LIMITATIONS: Nonrandom, unblinded treatment assignment was used, and follow-up data were missing for 20% of patients. Loss to follow-up was more common among patients who were less sick. Two uncontrolled interim analyses were conducted. CONCLUSIONS: Compared with no treatment, benznidazole treatment was associated with reduced progression of Chagas disease and increased negative seroconversion for patients presenting with nonacute disease and no heart failure. These observations indicate that a randomized, controlled trial should now be conducted. PMID- 16702589 TI - Brief communication: Glomerulonephritis in patients with hepatitis C cirrhosis undergoing liver transplantation. AB - BACKGROUND: Patients infected with hepatitis C virus (HCV) frequently develop renal failure after liver transplantation. OBJECTIVE: To describe renal histologic characteristics and concomitant clinical features in HCV-infected patients with end-stage cirrhosis. DESIGN: Case series. SETTING: Single-center liver transplant program in the United States. PATIENTS: 30 patients who received liver transplants for HCV-induced cirrhosis. INTERVENTION: Kidney biopsy during liver engraftment. MEASUREMENTS: Clinical data and laboratory tests of renal function within 6 months before liver transplantation. RESULTS: Twenty-five patients had immune-complex glomerulonephritis: membranoproliferative glomerulonephritis type 1 (n = 12), IgA nephropathy (n = 7), and mesangial glomerulonephritis (n = 6). Of these patients, 10 had normal serum creatinine levels, normal urinalysis results, and normal quantitative proteinuria. For 5 others, the only renal abnormality was an increased serum creatinine level. No patient had cryoglobulins in the blood or kidney. LIMITATIONS: This small observational study did not include patients with nonviral cirrhosis and did not document post-transplantation outcomes. CONCLUSIONS: Immune-complex glomerulonephritis was common in patients with end-stage HCV-induced cirrhosis and was often clinically silent. Its potential to cause renal failure after liver transplantation may be underappreciated. PMID- 16702591 TI - Narrative [corrected] review: bisphosphonates and osteonecrosis of the jaws. AB - Osteonecrosis of the jaws is a recently described adverse side effect of bisphosphonate therapy. Patients with multiple myeloma and metastatic carcinoma to the skeleton who are receiving intravenous, nitrogen-containing bisphosphonates are at greatest risk for osteonecrosis of the jaws; these patients represent 94% of published cases. The mandible is more commonly affected than the maxilla (2:1 ratio), and 60% of cases are preceded by a dental surgical procedure. Oversuppression of bone turnover is probably the primary mechanism for the development of this condition, although there may be contributing comorbid factors. All sites of potential jaw infection should be eliminated before bisphosphonate therapy is initiated in these patients to reduce the necessity of subsequent dentoalveolar surgery. Conservative debridement of necrotic bone, pain control, infection management, use of antimicrobial oral rinses, and withdrawal of bisphosphonates are preferable to aggressive surgical measures for treating this condition. The degree of risk for osteonecrosis in patients taking oral bisphosphonates, such as alendronate, for osteoporosis is uncertain and warrants careful monitoring. PMID- 16702590 TI - Systematic review: impact of health information technology on quality, efficiency, and costs of medical care. AB - BACKGROUND: Experts consider health information technology key to improving efficiency and quality of health care. PURPOSE: To systematically review evidence on the effect of health information technology on quality, efficiency, and costs of health care. DATA SOURCES: The authors systematically searched the English language literature indexed in MEDLINE (1995 to January 2004), the Cochrane Central Register of Controlled Trials, the Cochrane Database of Abstracts of Reviews of Effects, and the Periodical Abstracts Database. We also added studies identified by experts up to April 2005. STUDY SELECTION: Descriptive and comparative studies and systematic reviews of health information technology. DATA EXTRACTION: Two reviewers independently extracted information on system capabilities, design, effects on quality, system acquisition, implementation context, and costs. DATA SYNTHESIS: 257 studies met the inclusion criteria. Most studies addressed decision support systems or electronic health records. Approximately 25% of the studies were from 4 academic institutions that implemented internally developed systems; only 9 studies evaluated multifunctional, commercially developed systems. Three major benefits on quality were demonstrated: increased adherence to guideline-based care, enhanced surveillance and monitoring, and decreased medication errors. The primary domain of improvement was preventive health. The major efficiency benefit shown was decreased utilization of care. Data on another efficiency measure, time utilization, were mixed. Empirical cost data were limited. LIMITATIONS: Available quantitative research was limited and was done by a small number of institutions. Systems were heterogeneous and sometimes incompletely described. Available financial and contextual data were limited. CONCLUSIONS: Four benchmark institutions have demonstrated the efficacy of health information technologies in improving quality and efficiency. Whether and how other institutions can achieve similar benefits, and at what costs, are unclear. PMID- 16702592 TI - A framework for management of hepatitis C in prisons. AB - The prevalence of chronic hepatitis C virus (HCV) infection in prisons ranges from 12% to 31%. There are generally accepted--albeit still evolving--guidelines for identification and treatment of hepatitis C in the community. However, there is less agreement among health professionals caring for prisoners about best practices for identification, medical management, and treatment of hepatitis C. Inmates often lack health care before incarceration. In prisons, infected persons could be identified and the management of infection initiated; however, the high prevalence of HCV infection among prisoners would impose a disproportionate cost for hepatitis C care on the correctional system. The optimal solution is for prison and public health systems in the United States to jointly provide targeted HCV testing and standard-of-care hepatitis C medical management, treatment, and prevention programs to prison inmate populations. The authors report on a January 2003 meeting of experts in prison health, public health, hepatology, and infectious diseases and explore the clinical care, prevention, and collaboration needed to provide hepatitis C management in prisoners. PMID- 16702593 TI - Hepatitis C: a bitter harvest. PMID- 16702594 TI - The treatment of Chagas disease (South American trypanosomiasis). PMID- 16702595 TI - Health information technology: shall we wait for the evidence? PMID- 16702596 TI - Joan's education. PMID- 16702597 TI - Transformation: from medical student to patient. PMID- 16702598 TI - Socioeconomic status and mortality. PMID- 16702599 TI - Report of specific cardiovascular outcomes of the ADVANTAGE trial. PMID- 16702600 TI - Acute myocardial infarction associated with the serotonin syndrome. PMID- 16702601 TI - Conjugated linoleic acid inhibits osteoclast differentiation of RAW264.7 cells by modulating RANKL signaling. AB - Bone destruction is a pathological hallmark of several chronic inflammatory diseases, including rheumatoid arthritis, periodontitis, and osteoporosis. Inflammation-induced bone loss of this sort results from increased numbers of bone-resorbing osteoclasts. Numerous studies have indicated that conjugated linoleic acid (CLA) positively influences calcium and bone metabolism. Gene deletion studies have shown that receptor activator of nuclear factor-kappaB ligand (RANKL) is one of the critical mediators of osteoclastogenesis. In this report, we examine the ability of CLA to suppress RANKL signaling and osteoclastogenesis in RAW264.7 cells, a murine monocytic cell line. Treatment of these cells with RANKL activated nuclear factor-kappaB (NF-kappaB), and preexposure of the cells to CLA significantly suppressed RANKL-induced NF-kappaB activation, including phosphorylation of I-kappaBalpha, degradation of I kappaBalpha, and nuclear translocation of p65. RANKL induced osteoclastogenesis in these monocytic cells, and CLA inhibited RANKL-induced tumor necrosis factor alpha production and osteoclast differentiation, including osteoclast-specific genes such as tartrate-resistant acid phosphatase, cathepsin K, calcitonin receptor, and matrix metalloproteinase-9 expression and osteoclast-specific transcription factors such as c-Fos, nuclear factor of activated T-cells expression, and bone resorption pit formation. CLA also inhibited RANKL-induced activation of mitogen-activated protein kinase p38 but had little effect on c-Jun N-terminal kinase activation. Collectively, these data demonstrate for the first time that CLA inhibits osteoclastogenesis by modulating RANKL signaling. Thus, CLA may have important therapeutic implications for the treatment of bone diseases associated with enhanced bone resorption by excessive osteoclastogenesis. PMID- 16702602 TI - Efflux of sphingomyelin, cholesterol, and phosphatidylcholine by ABCG1. AB - Cholesterol and phospholipids are essential to the body, but an excess of cholesterol or lipids is toxic and a risk factor for arteriosclerosis. ABCG1, one of the half-type ABC proteins, is thought to be involved in cholesterol homeostasis. To explore the role of ABCG1 in cholesterol homeostasis, we examined its subcellular localization and function. ABCG1 and ABCG1-K120M, a WalkerA lysine mutant, were localized to the plasma membrane in HEK293 cells stably expressing ABCG1 and formed a homodimer. A stable transformant expressing ABCG1 exhibited efflux of cholesterol and choline phospholipids in the presence of BSA, and the cholesterol efflux was enhanced by the presence of HDL, whereas cells expressing ABCG1-K120M did not, suggesting that ATP binding and/or hydrolysis is required for the efflux. Mass and TLC analyses revealed that ABCG1 and ABCA1 secrete several species of sphingomyelin (SM) and phosphatidylcholine (PC), and SMs were preferentially secreted by ABCG1, whereas PCs were preferentially secreted by ABCA1. These results suggest that ABCA1 and ABCG1 mediate the lipid efflux in different mechanisms, in which different species of phospholipids are secreted, and function coordinately in the removal of cholesterol and phospholipids from peripheral cells. PMID- 16702603 TI - Type 2 immunity is controlled by IL-4/IL-13 expression in hematopoietic non eosinophil cells of the innate immune system. AB - Nippostrongylus brasiliensis infection and ovalbumin-induced allergic lung pathology are highly interleukin (IL)-4/IL-13 dependent, but the contributions of IL-4/IL-13 from adaptive (T helper [Th]2 cells) and innate (eosinophil, basophils, and mast cells) immune cells remain unknown. Although required for immunoglobulin (Ig)E induction, IL-4/IL-13 from Th2 cells was not required for worm expulsion, tissue inflammation, or airway hyperreactivity. In contrast, innate hematopoietic cell-derived IL-4/IL-13 was dispensable for Th2 cell differentiation in lymph nodes but required for effector cell recruitment and tissue responses. Eosinophils were not required for primary immune responses. Thus, components of type 2 immunity mediated by IL-4/IL-13 are partitioned between T cell-dependent IgE and an innate non-eosinophil tissue component, suggesting new strategies for interventions in allergic immunity. PMID- 16702604 TI - Vascular endothelial growth factor is an important determinant of sepsis morbidity and mortality. AB - Sepsis, the systemic inflammatory response to infection, is a leading cause of morbidity and mortality. The mechanisms of sepsis pathophysiology remain obscure but are likely to involve a complex interplay between mediators of the inflammatory and coagulation pathways. An improved understanding of these mechanisms should provide an important foundation for developing novel therapies. In this study, we show that sepsis is associated with a time-dependent increase in circulating levels of vascular endothelial growth factor (VEGF) and placental growth factor (PlGF) in animal and human models of sepsis. Adenovirus-mediated overexpression of soluble Flt-1 (sFlt-1) in a mouse model of endotoxemia attenuated the rise in VEGF and PlGF levels and blocked the effect of endotoxemia on cardiac function, vascular permeability, and mortality. Similarly, in a cecal ligation puncture (CLP) model, adenovirus-sFlt-1 protected against cardiac dysfunction and mortality. When administered in a therapeutic regimen beginning 1 h after the onset of endotoxemia or CLP, sFlt peptide resulted in marked improvement in cardiac physiology and survival. Systemic administration of antibodies against the transmembrane receptor Flk-1 but not Flt-1 protected against sepsis mortality. Adenovirus-mediated overexpression of VEGF but not PlGF exacerbated the lipopolysaccharide-mediated toxic effects. Together, these data support a pathophysiological role for VEGF in mediating the sepsis phenotype. PMID- 16702605 TI - Feruloyl esterase: a key enzyme in biomass degradation. AB - Feruloyl esterase forms a part of the enzyme complex that acts collectively and synergistically to completely hydrolyze xylan to its monomers. The enzyme has found potential uses in a wide variety of applications of interest to the agrifood and pharmaceutical industries. This review describes the enzymology of feruloyl esterases involved in xylan degradation. The occurrence of feruloyl esterases in various microorganisms and their physiochemical properties are presented. The nature of the enzyme substrates and products, the role of synergistic interactions with xylanases and other accessory enzymes, as well as the sequence-structure relating to the reaction mechanism are emphasized. PMID- 16702606 TI - Modeling and optimization of biopolymer (polyhydroxyalkanoates) production from ice cream residue by novel statistical experimental design. AB - Polyhydroxyalkanoates (PHAs) are thermoplastic polyesters synthesized by Ralstonia eutropha and other bacteria as a form of intracellular carbon and energy storage and are accumulated as lipid inclusions in the cytoplasm of these bacteria. The modeling and optimization of PHA production by fermentation from industrial waste (ice cream residue) was studied by employing statistical experimental design methods. A series of iterative experimental designs was used to find optimal factor conditions (medium components and fermentation process time) in the order of fractional factorial design, path of steepest ascent, and full factorial augmented with axial design (rotational central composite design). An optimal range characterized by lipid (15 mg/mL) and % lipid (88%) values was found and further investigated to verify the optimal conditions for PHA production from ice cream (56.68 mL of ice cream or 56.68% ice cream in water [v/v], 5.03 mL of buffer, 1 mL of mineral salts solution, 100 muL of trace element solution, 100 mL of seed culture, and 213.76 h of fermentation time). PMID- 16702607 TI - Expression, refolding, and characterization of GFE peptide-fused human interferon alpha2a in Escherichia coli. AB - Interferon-alpha2a (IFN-alpha2a) has been used for the treatment of various viral infections and cancers for many years. However some untolerable side effects have limited its application in some aspects. To evaluate whether or not an oligopeptide containing GFE motif can home human IFN-alpha2a to specific tissues, a fusion gene was constructed by fusing the coding sequence of GFE peptide (CGFECVRQCPERC), which was screened from phage display peptide library, to the 3' end of human IFN-alphaa gene by recombinant DNA technique. Fusion protein rhIFN alpha2a-GFE was expressed in Escherichia coli as inclusion bodies using a T7 RNA polymerase expression system, pET-22b, refolded through dialysis and purified to homogeneity to >95% of purity by affinity chromatography. Characterization by sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotting demonstrated the authenticity of the fusion protein. Purified rhIFN-alpha2a-GFE was found to be functionally active in terms of its antiviral activity for about 2.5 x 108 IU/mg in vitro. Yields of the purified fusion protein were about 200 mg/L of culture medium. Tissue distribution assay in mouse showed that at 30 min IFN-alpha2a could be enriched sevenfold higher in lung in the targeted IFN group of mice than in the standard IFN group of mice, and last for a long time. At 1 h, IFN-alphaa in the targeted IFN group was still 4.02-fold higher than that in the standard group. This confirmed that GFE peptide has the ability to selectively deliver its fusion partner IFN-alpha2a to lungs. The results also showed that the IFN-alpha2a-GFE could be specifically enriched in kidney and liver. Its distribution in kidney was concordant with the finding of GFE receptor, MDP, in kidney. However, the IFN-alpha2a-GFE in liver may imply some significance in pharmacology and toxicology. PMID- 16702608 TI - Production of beta-galactosidase by Trichoderma reesei FTKO-39 in wheat bran: partial purification of two isozymes. AB - Trichoderma reesei FTKO-39 grown at 35 degrees C for 5 d on wheat bran supplemented with MgCl2 and lactose as the carbon source produced two isozymes of beta-galactosidase: BGT I and BGT II. These isozymes were partially purified on a DEAE-Trisacryl column. Both BGT I and BGT II fractions exhibited optimum activity at 65 degrees C, but the pH optima were 4.0 and 6.5, respectively. The isozymes also showed similar thermal stability. However, BGT I was more stable than BGT II in a pH range of 3.0-10.0. At least two different beta-galactosidases are produced by T. reesei, as revealed by the two bands seen on a 6% polyacrylamide gel stained for activity. PMID- 16702609 TI - Analysis of performance of an anaerobic sequencing batch reactor submitted to increasing organic load with different influent concentrations and cycle lengths. AB - The performance of an anaerobic sequencing batch reactor (ASBR) was assessed when submitted to increasing organic load with different influent concentrations and cycle lengths. The 5-L mechanically stirred (75 rpm) ASBR contained 2 L of granular biomass and treated 2 L of synthetic wastewater per cycle. Volumetric organic loads (VOLs) from 0.66 to 2.88 g of chemical oxygen demand (COD)/(L x d) were applied by using influent concentrations from 550 to 3,600 mg of COD/L in 8- and 12-h cycles. Reactor stability was maintained for VOLs from 0.66 to 2.36 g of COD/(L x d), with organic matter removal efficiencies for filtered samples (epsilonF) between 84 and 88%. For VOLs from 0.78 to 2.36 g of COD/(L x d) at an influent concentration of 2,000 mg of COD/L, when cycle length was reduced from 12 to 8 h, epsilonF did not vary, yet showed a very distinct behavior from the other conditions. In addition, two operation strategies were studied for VOLs with approximately similar values of 2.36 and 2.08 g of COD/(L x d). One involved operation with an influent concentration of 2,000 mg of COD/L and an 8-h cycle, whereas the other involved an influent concentration of 2,600 mg of COD/L and a 12-h cycle. Only the former resulted in system stability and efficiency. These results indicate that besides organic load, influent concentration and cycle length play a significant role in ASBR systems. PMID- 16702610 TI - Treatment of liver metastases from colorectal cancer. AB - In recent years several new local as well as systemic treatment options have become available for patients with advanced colorectal cancer. A survey among Dutch hospitals revealed considerable differences in the use of diagnostic and therapeutic strategies. Radiofrequency ablation is a promising technique that is currently being investigated in a randomised trial. The role of adjuvant chemotherapy in patients with resectable liver metastases and of neoadjuvant chemotherapy in patients with nonresectable liver metastases has not been clearly established yet. The current status of local and systemic treatment options for colorectal cancer liver metastases is reviewed. PMID- 16702611 TI - Extranodal marginal zone (MALT) lymphoma in common variable immunodeficiency. AB - We describe two patients with common variable immunodeficiency (CVID) who developed extranodal marginal zone lymphoma (formerly described as mucosa associated lymphoid tissue lymphoma or MALT lymphoma). One patient, with documented pernicious anaemia and chronic atrophic gastritis with metaplasia, developed a Helicobacter pylori-positive extranodal marginal zone lymphoma in the stomach. Three triple regimens of antibiotics were necessary to eliminate the H. pylori, after which the lymphoma completely regressed. Patient B had an H. pylori negative extranodal marginal zone lymphoma of the parotid gland, which remarkably regressed after treatment with clarithromycin. Reviewing the literature, we found eight cases of extranodal marginal zone lymphoma complicating CVID, but probably many more cases labelled as non-Hodgkin's lymphoma are hidden in the literature. Until more data are available on the predictive value of noninvasive screening for pathology of the stomach, we recommend endoscopy to assess the gastric status in CVID patients in order to detect these malignancies at an early stage. Elimination of H. pylori infection is the treatment of choice in Helicobacter positive extranodal marginal zone lymphoma. The possibility of elimination failure, most probably due to frequent and prolonged exposure to antibiotics in this patient group, should be taken into account. Treatment with antibiotics in Helicobacter-negative extranodal marginal zone lymphoma must be considered. PMID- 16702613 TI - Imaging and treatment of patients with colorectal liver metastases in the Netherlands: a survey. AB - BACKGROUND: Clinical experience has highlighted the absence of a uniform approach to the management of patients with colorectal liver metastases in the Netherlands. METHODS: A written survey on the diagnosis and treatment of patients with colorectal liver metastases was sent to all 107 chairmen of oncology committees in each hospital. Questions were asked concerning: specialists involved in decision-making, availability and existence of guidelines and meetings, factors that needed to be improved, information regarding the diagnostic work-up of liver metastases, detailed techniques of ultrasonography (US), computed tomography (CT), magnetic resonance imaging (MRI) and positron emission tomography (PET), factors influencing resectability, types of surgery performed, the use of (neo)adjuvant chemotherapy, portal vein embolisation performance, considering isolated hepatic perfusion (IHP) or local ablation as treatment options, actual performance of local ablation and the use of systemic as well as regional chemotherapy. RESULTS: Response rate was 68% (73/107). Specialists involved in the management were mostly surgeons (70), medical oncologists (66) and radiologists (42). Factors that needed to be improved, as indicated by responders, were the absence of 1) guidelines; 2) registration of patients and 3) guidelines for radiofrequency ablation (RFA). Diagnostic work-up of synchronous liver metastases occurred in 71 hospitals, (by US in 69 and by CT in 2). For the work-up of metachronous liver metastases, US was used as initial modality in 14, CT in 2 hospitals, and 57 hospitals used one or the other (mainly US). As additional modality, CT was performed (71) and to a lesser extent MRI (38) or PET (22). Diagnostic laparoscopy and biopsy were performed incidentally. The choice for an imaging modality was mostly influenced by the literature, and to a lesser extent by the availability and by costs, personnel and waiting lists. Substantial variation exists in the US, CT, MRI and PET techniques. The absence of extrahepatic disease and the clinical condition were considered as the most important factors influencing resectability. Surgery was performed in 30 hospitals; hemihepatectomy in 25, segment resection in 27, multisegment resection in 23, wedge excision in 27 and combination of resection and RF A in 18 institutions. In 52 hospitals (neo)adjuvant chemotherapy was administrated to improve surgical results, partly (35%) in trials. In nine hospitals portal vein embolisation was performed, with the volume of the remnant liver as the most important factor. Local ablative techniques were considered as a treatment option in 48 hospitals and actually performed in 16 hospitals, without clearly defined indications. Experimental IHP was considered a treatment option by 45 (62%) responders, irrespective whether this treatment was available at their centre. Patients with extensive metastases received systemic chemotherapy in all 73 hospitals and regional chemotherapy in ten hospitals. CONCLUSION: This survey shows substantial variation in the diagnostic and therapeutic work-up of patients with colorectal liver metastases. This variation reflects either under- or over utilisation of diagnosis and treatment options. Evidence-based guidelines taking into account the available evidence, experience and availability can solve this variation. PMID- 16702612 TI - Capecitabine, epirubicin and cisplatin in the treatment of oesophagogastric adenocarcinoma. AB - BACKGROUND: Inoperable or metastatic oesophagogastric adenocarcinoma has a poor prognosis. From the many different chemotherapeutic regimens used in the past, a combination of epirubicin, cisplatin and continuous 5-fluorouracil infusion (ECF) showed a consistent response rate of +/- 50% with acceptable toxicity. Continuous 5-FU infusion may be replaced by oral fluoropyrimidines. Here we evaluate treatment with epirubicin and cisplatin combined with oral capecitabine (ECC), replacing intravenous 5-FU infusion. METHODS: Retrospectively, we analysed 23 consecutive patients who were treated with epirubicin, cisplatin and oral capecitabine for inoperable or metastatic oesophagogastric adenocarcinoma during 2002 and 2003. RESULTS: The overall response rate was 57%; another 26% achieved stable disease and only 17% had progressive disease. The median duration of response was 6.4 months; the median survival was 9.0 months. Previously treated patients (n=10) had a significantly worse overall response rate (20%) compared with previously untreated patients (85%). A nonsignificant difference in median survival was found between these groups (3.9 vs 9.8 months in previously treated vs untreated patients). An acceptable incidence of grade 3 and 4 toxicity was found. CONCLUSION: Capecitabine in combination with epirubicin and cisplatin is an effective and safe alternative to ECF, without the risks of a continuous venous access. PMID- 16702614 TI - An unusual cause of a cerebral tumour in a young patient. Behcet's disease. PMID- 16702615 TI - Serious envenomation after a snakebite by a Western bush viper (Atheris chlorechis) in the Netherlands: a case report. AB - Venomous snakebites are a rarity in the Netherlands. In this report we describe the case of a 26-year-old male amateur snakekeeper who was bitten in his left index finger by a Western bush viper (Atheris chlorechis). His clinical condition deteriorated rapidly with acute renal failure and considerable blood loss due to coagulopathy. Antidote was not readily available and was finally supplied by a zoo in Antwerp, Belgium. One day after admission the blood loss diminished. PMID- 16702616 TI - A case of abdominal mesothelioma diagnosed by indium-III leucocyte scintigraphy. AB - We present a case of peritoneal mesothelioma that presented with fever of unknown origin and an elevation in the inflammatory parameters. Radiological imaging did not reveal a diagnosis. Because of tumour-associated inflammatory activity, indium-III leucocyte scintigraphy enabled us to establish a diagnosis. To our knowledge, the use of indium-III leucocyte scintigraphy in peritoneal mesothelioma has not been reported previously. PMID- 16702617 TI - Acute dystonic reaction to metoclopramide in patients carrying homozygous cytochrome P450 2D6 genetic polymorphisms. AB - BACKGROUND: Extrapyramidal syndromes (EPS) are clinically relevant side effects of metoclopramide which are often not anticipated. PATIENTS AND METHODS: Two patients who received metoclopramide developed an acute dystonic reaction. Symptoms disappeared after biperiden or trihexyphenidyl were given. Molecular analysis of the CYP2D6 gene was performed using a PCR-based method. RESULTS: Both patients were homozygous for inactive CYP2D6 alleles (CYP2D6*4/*4 and CYP2D6*4/*5), which are associated with slow drug metabolism. CONCLUSION: Metoclopramide-induced acute dystonic reactions may occur in patients carrying a CYP2D6 genetic polymorphism. PMID- 16702618 TI - Asymmetric dimethylarginine (ADMA) as a target for pharmacotherapy. AB - Asymmetric dimethylarginine (ADMA) is synthesized during the methylation of protein arginine residues by protein arginine methyltransferases (PRMT) and is released during proteolysis. ADMA is a competitive inhibitor of nitric oxide synthase and may decrease NO availability. ADMA is eliminated by renal excretion or is metabolized by dimethylarginine dimethylaminohydrolase (DDAH) to citruline and dimethylamine. Two other endogenous methylarginines are also synthesized by PRMT: N-monomethyl-L-arginine (L-NMMA) and symmetric dimethylarginine (SDMA). L NMMA inhibits NO synthase but its concentrations in circulation are much lower than ADMA whereas SDMA is inactive. Plasma concentration of ADMA is markedly increased in patients with chronic renal failure and moderately increased in patients with many other diseases including hyperlipidemia, diabetes mellitus, arterial hypertension, hyperhomocysteinemia and heart failure. The increased concentration of ADMA is positively correlated with markers of atherosclerosis, such as carotid artery intima-media thickness and has a predictive value for acute cardiovascular events in prospective studies. Angiotensin-converting enzyme inhibitors, angiotensin AT1 receptor antagonists, vitamin E and, according to some studies, estrogens used in hormonal replacement therapy reduce plasma ADMA concentration, which may contribute to their beneficial effect on NO synthesis and endothelial function. However, in some states associated with excess of NO, such as septic shock or excitotoxic neuronal injury ADMA may be protective by limiting toxic effect of high concentrations of NO. This article reviews the effect of pharmacotherapy on ADMA metabolism and its possible clinical implications. PMID- 16702619 TI - Vitamin E in the prevention of ischemic heart disease. AB - Ischemic heart disease (IHD) has now assumed a global dimension. It still remains one of the major health problems not only in the advanced countries, but also, is becoming a serious health issue in the developing and the economically weaker countries. Apart from other factors, changing economic scenario, stress and strain in daily life as well as altered dietary habits in the populations appear responsible for the increased incidence of cardiovascular disease (CVD). The treatment modalities, invasive, non-invasive and pharmacological are economically no dearer, even to population of affluent countries. Likewise, treatment costs of serious cardiovascular diseases are becoming difficult to be borne by population of the developing nations. Prevention of IHD would be a better way to protect the population from physical and economic disaster. The current article comprehensively describes the relation between oxidative stress and cardiac disease, explains the direct effect of reactive oxygen species on cardiac function and projects how the use of vitamin E can be of benefit in the prevention of IHD with concluding remarks highlighting the need for inclusion of a fruit and vegetable rich diet and regular exercise to keep the dearer heart active and healthy. PMID- 16702620 TI - Functional and molecular identification of 5-hydroxytryptamine receptors in rabbit pulmonary artery: involvement in complex regulation of noradrenaline release. AB - The rabbit pulmonary artery was used to examine whether presynaptic serotonin (5 HT) receptors modulate noradrenaline (NA) release also in this blood vessel and to confirm their presence with molecular biology techniques. Arteries preincubated with [3H]NA were superfused in the presence of the alpha2 adrenoceptor blocker rauwolscine and the effects of 5-HT receptor ligands on the electrically evoked 3H overflow were determined. The 5-HT4 receptor agonist cisapride inhibited 3H overflow in a manner sensitive to blockade by atropine. The 5-HT1B/1D receptor agonist 5-carboxamidotryptamine inhibited 3H overflow only in the presence of atropine. The 5-HT4 and 5-HT1B/1D receptor agonists 5-HT and 5 methoxytryptamine reduced 3H overflow in the absence and presence of atropine, and this effect was blocked by methiothepin, a non-selective 5-HT receptor antagonist, in the presence of atropine. PCR with cDNAs derived from reverse transcribed blood vessel mRNA suggested the expression of the 5-HT1B, 5-HT1D and 5-HT4 receptors, the latter being highly homologous to the human one. In conclusion, the cholinergic nerves are endowed with excitatory 5-HT4 receptors mediating release of acetylcholine which, in turn, activates muscarine receptors on the sympathetic nerves leading to inhibition of NA release. Blockade of the presynaptic muscarinic receptors involved is necessary to disclose an inhibition of NA release via 5-HT1B/1D receptors. Taking results reported in the literature into account, the 5-HT1D and 5-HT4 receptors identified by molecular biology techniques probably are located predominantly on the noradrenergic and cholinergic neurons, respectively. PMID- 16702621 TI - Effects of cannabinoids on the anxiety-like response in mice. AB - Several pieces of anatomical, biochemical and pharmacological evidence indicate that the endocannabinoid system via CB1 receptors is implicated in the control of emotional behavior. However, previous studies have reported unclear and contradictory results concerning the role of cannabinoids in anxiety. The aim of the present study was to examine the influence of the cannabinoid agonist WIN 55,212-2 (1 and 5 mg/kg), the CB1 antagonist AM 281 (1, 2 and 4 mg/kg), the inhibitor of anandamide hydrolysis AACOCF3 (1 and 4 mg/kg) and the inhibitor of anandamide transporter AM404 (1 and 4 mg/kg) on the anxiety-like response in mice in the light/dark box test. WIN 55,212-2 (5 mg/kg) induced the anxiogenic-like effect accompanied by motor inhibition, AACOCF3 (4 mg/kg) induced the selective anxiolytic-like effect, whereas AM404 and AM281 were without effect. Pretreatment with AM281 (2 mg/kg) blocked the anxiogenic-like and sedative responses induced by WIN 55, 212-2, as well as the anxiolytic-like effect of AACOCF3. These results support the hypothesis that the endocannabinoid system is involved in the regulation of anxiety-like behavior, and also suggest that the inhibitors of anandamide hydrolysis might be potential anxiolytic drugs. PMID- 16702622 TI - Impact of aromatic substitution on the anticonvulsant activity of new N-(4 arylpiperazin-1-yl)-alkyl-2-azaspiro[4,5]decane-1,3-dione derivatives. AB - A series of N-[(4-arylpiperazin-1-yl)-alkyl]-2-azaspiro[4.5]decane-1,3-dione derivatives were synthesized and evaluated for their anticonvulsant and neurotoxic properties. The main modifications to that series of compounds consisted in the introduction of an aromatic area to the cyclohexane ring as a flexible fragment with conformational freedom (1a-h), or as a rigidified skeleton (2a-h). Except for N-[3-(4-phenylpiperazin-1-yl)-propyl]-8-phenyl-2-aza spiro[4.5]decane-1,3-dione derivative (1e), all the other compounds displayed anticonvulsant activity in the MES test, but some of them (1c, 2f and 2g) were found to be neurotoxic at a dose of 30 mg/kg, irrespective of their activity. The most potent and relatively weakly neurotoxic analogues of that series, i.e. N-[2 {4-(3-chlorophenyl)-piperazin-1-yl}-ethyl]-[7,8-f]benzo-2-aza-spiro[4.5]decane 1,3-dione (2c) and N-[3-{4-(3-trifluoromethylphenyl)-piperazin-1-yl}-propyl]-[7,8 f] benzo-2-aza-spiro[4.5]decane-1,3-dione (2h) had ED50 values of 205 mg/kg (2c) and 23 mg/kg (2h) respectively, in the MES-test in mice, and showed higher protection than magnesium valproate (ED50 = 211 mg/kg), used as a standard substance. PMID- 16702623 TI - Bismuth increases hydroxyl radical-scavenging activity of histamine H2-receptor antagonists. AB - The effects of histamine H2-receptor antagonists, alone or in a combination with bismuth, on *OH-provoked degradation of deoxyribose were studied. The histamine H2-receptor antagonists (cimetidine, ranitidine and roxatidine), themselves decreased the deoxyribose damage in Fenton-type systems. In combinations with bismuth, their inhibitory effect in Fenton system (Fe(III)/ascorbic acid + H2O2 was stronger. Moreover, unlike F(III) and Cu(II), which in the presence of ascorbic acid + H2O2 led to an increase in the *OH formation (deoxyribose damage), Bi(III) showed an opposite effect. The present results are interpreted in view of a better ( )OH scavenging activity of bismuth complexes of histamine H2-receptor antagonists as compared to that of the corresponding drugs. These findings might be one more explanation why bismuth salts, in combination with acid-reducing agents, are more effective anti-ulcer agents. PMID- 16702624 TI - Effects of diphenhydramine and famotidine on lipid peroxidation and activities of antioxidant enzymes in different rat tissues. AB - The potential antioxidant activity of diphenhydramine (histamine H1-receptor antagonist) and famotidine (histamine H2 receptor antagonist) was studied. Diphenhydramine inhibited the spontaneous, Fe(II)-induced and Fe(II)/ascorbate induced lipid peroxidation, while famotidine showed a biphasic concentration dependent effect on spontaneous lipid peroxidation (a stimulation by 1 mM and an inhibition by 5 mM) and increased Fe(II)-induced- and inhibited Fe(II)/ascorbate induced lipid peroxidation in the rat liver and brain. Both drugs decreased *OH provoked deoxyribose degradation in Fenton-type systems and inhibited O2- provoked reduction of nitro-blue tetrazolium and ferrycytochrome C, but famotidine effect was stronger than that of diphenhydramine. The significant famotidine-induced inhibition of nitro-blue tetrazolium reduction might be underlain by the stimulation of superoxide dismutase activity. Famotidine and diphenhydramine did not alter the catalase activity in all tissue preparations, except for its concentration of 5 mM (a complete inhibition). The present results suggest a beneficial effect of histamine H1 and H2-blockers, especially famotidine, as antioxidants and/or metal chelators, which might be an additional explanation of their therapeutic action. PMID- 16702625 TI - Prevention of 1,2-dimethylhydrazine-induced circulatory oxidative stress by bis 1,7-(2-hydroxyphenyl)-hepta-1,6-diene-3,5-dione during colon carcinogenesis. AB - We have performed this study to investigate the modulatory effect of bis-1,7-(2 hydroxyphenyl)-hepta-1,6-diene-3,5-dione, a bisdemethoxy curcumin analog (BDMCA) on circulatory lipid peroxidation (LPO) and antioxidant status during 1,2 dimethylhydrazine (DMH)-induced colon carcinogenesis in male Wistar rats. The effects were compared with that of the reference drug, curcumin. Increased tumor incidence as well as enhanced LPO in the circulation of tumor bearing rats was accompanied by a significant decrease in the level of reduced glutathione and activities of glutathione peroxidase (GPx), glutathione S-transferase (GST), superoxide dismutase (SOD) and catalase (CAT). Intragastric administration of BDMCA or curcumin to DMH-treated rats significantly decreased colon tumor incidence and the circulatory LPO, with simultaneous enhancement of GSH content and GPx, GST, SOD and CAT activities. We report that BDMCA exert its chemopreventive effect by decreasing the colon tumor incidence as well as by modulating circulatory oxidative stress in DMH-treated rats through its influence on LPO and antioxidant status. The effects of BDMCA were comparable with that of the reference compound curcumin, a well known anticarcinogen and antioxidant. Thus, it would be suggested that the methoxy group is not responsible for the beneficial effects, however, the terminal phenolic moieties or the central 7 carbon chain may play a role. PMID- 16702626 TI - Immunosuppressory activity of an isoxazolo[5,4-e]triazepine-compound RM-33 II. Effects on the carrageenan-induced inflammation. AB - The aim of this investigation was to evaluate effectiveness of RM-33, a new isoxazolotriazepine, in the model of carrageenan-induced inflammation in rats. Wistar rats were pretreated with intraperitoneal (ip) or oral (po) doses of RM 33, at daily doses ranging from 250 to 1000 microg, administered 1-3 days before elicitation of the carrageenan reaction. We showed that both routes of RM-33 administration were effective in significantly diminishing the footpad edema. The effects were dose-dependent and better pronounced at the ip administration of the compound. We found a lower production of tumor necrosis factor alpha (TNF-alpha) by mitogen-stimulated splenocytes isolated from rats pretreated with RM-33 and injected with carrageenan, as well as lower serum TNF-alpha levels in these rats, as compared to the respective control. Histological analysis of the skin reaction site revealed that in the rats pretreated with RM-33, the carrageenan-induced inflammation was reduced, as reflected by a lesser damage of mast cells, smaller infiltration by macrophages and a diminished edema of the connective tissue. Together with our previous data, indicating the antagonistic action of RM-33 in the adjuvant-induced footpad inflammation in mice, the present results confirm the anti-inflammatory activity of RM-33 compound. PMID- 16702627 TI - Anticonvulsant effect of amiloride in pentetrazole-induced status epilepticus in mice. AB - Inhibition of sodium hydrogen exchangers (NHE) has been shown to diminish seizure activity in various in vitro and in vivo models of epilepsy. In the present study, we examined the effect of amiloride, a sodium hydrogen exchanger inhibitor, against pentetrazole (PTZ)-induced status epilepticus (SE). The study was conducted in mice and status epilepticus was induced by administering ip 50 mg/kg of phenytoin followed 2 hour later by PTZ, 100 mg/kg sc. Amiloride produced dose-dependent protection against PTZ-induced SE. PMID- 16702631 TI - Results of survey of national influenza pandemic preparedness in Europe. PMID- 16702628 TI - Search for drugs of the combined anti-inflammatory and anti-bacterial properties: 1-methyl-N'-(hydroxymethyl)nicotinamide. AB - It has already been reported that 1-methylnicotinamide (MNA+), a primary metabolite of nicotinamide (vitamin B3), possesses remarkable anti-inflammatory properties [3]. This communication shows that 1-methyl-N' (hydroxymethyl)nicotinamide (MNAF+ can be regarded as MNA+ precursor able to release simultaneously formaldehyde. Therefore, MNAF+ can be viewed as a candidate for drug with the combined anti-inflammatory and anti-bacterial properties. PMID- 16702632 TI - Large ongoing rubella outbreak in religious community in the Netherlands since September 2004. PMID- 16702633 TI - Two cases of lymphogranuloma venereum (LGV) in homosexual men in Stockholm. PMID- 16702634 TI - Burkholderia pseudomallei infections in Finnish tourists injured by the December 2004 tsunami in Thailand. PMID- 16702635 TI - REPLACE: a European project on plants, their extracts and other natural alternatives to antimicrobials in animal feeds. PMID- 16702637 TI - Occupationally acquired HIV: international reports to December 2002. PMID- 16702636 TI - Nationwide outbreak of Salmonella enterica serotype Agona infections in infants in France, linked to infant milk formula, investigations ongoing. PMID- 16702638 TI - Hepatitis B in northwest Russia and the Nordic and Baltic countries: recent trends and prevention activities. PMID- 16702639 TI - Hepatitis B transmission in care homes linked to blood glucose monitoring, Belgium and United States. PMID- 16702640 TI - Large outbreak of Salmonella Enteritidis in North East London. PMID- 16702641 TI - Avian influenza--update on recent situation in South East Asia. PMID- 16702642 TI - Five case histories of tularaemia infection in Oppland and Hedmark Counties, Norway. PMID- 16702643 TI - A nationwide outbreak of Salmonella Bovismorbificans PT24, Germany, December 2004 March 2005. PMID- 16702644 TI - World Stop TB Day 2005: tuberculosis care providers and monitoring of treatment outcome in Europe. PMID- 16702645 TI - Decline in measles in WHO European Region but rubella remains high. PMID- 16702646 TI - Health complaints linked to surface water recreation in The Netherlands, 2000 2003. PMID- 16702647 TI - Occurrence of ticks infected by tickborne encephalitis virus and Borrelia genospecies in mountains of the Czech Republic. PMID- 16702648 TI - Preventing and controlling disease outbreaks in a complex emergency situation: discussion of the tsunami aftermath. PMID- 16702649 TI - AIDS in Africa: three scenarios in 2025. PMID- 16702650 TI - Mortality and causes of death in South Africa, 1997-2003: findings from death notifications. PMID- 16702651 TI - A Novel Technique for Accurate Intensity Calibration of Area X-ray Detectors at Almost Arbitrary Energy. AB - A novel intensity uniformity calibration method for area X-ray detectors is described. In diffraction experiments, amorphous lithium glass plates, containing doping elements chosen for their K edges just below the energy of the main beam, replace the crystallographic samples for the calibration measurement. The fluorescent emission excited by the X-ray beam is almost isotropic. It has exactly the same geometry as the diffracted radiation, and can be obtained at the same wavelength by proper selection of the element and excitation energy. A sample 2theta scan allows the emission distribution as a function of angle to be characterized with an accuracy of a fraction of a percent. This allows a flat field correction of similar accuracy. The quality of crystallographic data collected with an X-ray image intensifier/CCD detector was significantly improved by flat-field correction using an Sr-doped lithium tetraborate glass. This technique can be applied to X-ray energies from 5 to 50 KeV; the calibration sample is small, stable and easily handled. PMID- 16702652 TI - A Novel X-ray Diffractometer to Study the Texture of Materials. AB - An X-ray energy-dispersive diffraction technique to study the texture of materials using synchrotron radiation has been developed. The design and commissioning of the diffraction instrument are described. The technique was first applied to study a drawn-wire aluminum sample which has a well known deformation texture. To demonstrate its capability further, results obtained from an erbium evaporated thin film are also presented. PMID- 16702653 TI - Structure Solution of Azurin II from Alcaligenes xylosoxidans using the Laue Method: Possibility of Studying In Situ Redox Changes using X-rays. AB - We have recently demonstrated that X-rays can be used for changing the redox states of the metal centre in metalloproteins [Murphy et al. (1995). J. Synchrotron Rad. 2, 64-69]. The possibility of using the Laue method for studying the structural changes associated with such X-ray-induced reactions is explored by applying the method to the structure determination of a new azurin (hereafter referred to as azurin II) from the denitrifying bacterium Alcaligenes xylosoxidans. Laue X-ray diffraction data of azurin II were collected at station 9.7 of the SRS Daresbury. Three diffraction patterns were recorded on film packs at three different crystal orientations. The data were processed using the Daresbury Laue Software Suite to give 2224 independent single reflections (R(merge) = 0.136) in the wavelength range 0.36-1.40 A. The data completeness was 44% at 2.55 A resolution. Phase determination for the data was undertaken using the molecular-replacement method; the top peak was chosen in both the rotation function and the subsequent translation function. This solution agreed well with the molecular-replacement solution achieved independently using monochromatic data. The electron-density map showed reasonably good agreement with the model and the copper site was readily recognizable as it had the highest density. To see if the electron-density map could be improved, ;the doublets in the diffraction data were then deconvoluted. This added 26% data in the region infinity-2d(min) resulting in an improvement in the data completeness to 50% and thus in improved continuity of the electron-density map. The quality of these maps is discussed from the point of view of the suitability of this approach for studying redox-induced structural changes. PMID- 16702654 TI - Hydrogen-Atom Ordering in Lead Hydrogen Arsenate Monitored by Neutron Time-of Flight Laue Diffraction using Single-Frame Methods. AB - The ordering of the hydrogen atom in a single crystal of lead hydrogen arsenate in the temperature range 320 to 80 K is described. The details of the ordering at small temperature steps are determined from single frames of data collected using the neutron time-of-flight Laue method. The constrained refinements obtained from these data yield lower precision than full data sets but allow the ordering to be characterized rapidly and reliably. This is the first example of the use of this single-frame method of monitoring such phase transitions. The approach used here with neutrons, stimulated in part by synchrotron Laue diffraction, will be of interest for all time-resolved experiments with synchrotron radiation or neutrons. PMID- 16702655 TI - MAD Phasing Strategies Explored with a Brominated Oligonucleotide Crystal at 1.65A Resolution. AB - The crystal structure of a brominated oligonucleotide d(CGCG(Br)CG), chemical formula C(114)N(48)O(68)P(10)Br(2), has been analysed by multiwavelength anomalous dispersion (MAD) methods. The oligonucleotide crystallizes in space group P2(1)2(1)2(1) with a = 17.97, b = 30.98, c = 44.85 A, alpha = beta = gamma 90 degrees . Data to a resolution of 1.65 A were collected at four wavelengths about the K-absorption edge of the bromine atom (lambda(1) = 0.9323 A, a reference wavelength at the long-wavelength side of the edge; lambda(2) = 0.9192 A, at the absorption-edge inflection point; lambda(3) = 0.9185 A, at the ;white line' absorption maximum; lambda(4) = 0.8983 A, a reference wavelength at the short-wavelength side) using synchrotron radiation at Station PX9.5, SRS, Daresbury. Multiwavelength data could be collected on a single-crystal as the sample was radiation stable. Anomalous and dispersive Patterson maps were readily interpretable to give the bromine anomalous scatterer positions. Phase calculations to 1.65 A, resolution, using all four wavelengths, gave a figure of merit of 0.825 for 2454 reflections. The electron-density map was readily interpretable showing excellent connectivity for the sugar/phosphate backbone and each base was easily characterized. The two nucleotide strands paired up as expected in an antiparallel Watson-Crick-type manner. The structure was refined to 1.65 A using all the data (R-factor = 17.0% based on 3151 reflections, with a data-to-parameter ratio of 2.6). In addition to the four-wavelength analysis, a variety of other phasing strategies, and the associated quality of the resulting electron-density maps, were compared. These included use of either of the reference wavelength data sets in the two possible three-wavelength phasing combinations to assess their relative effectiveness. Moreover, the time dependence upon measuring the Bijvoet differences and its effect upon phasing was also investigated. Finally, the use of only two wavelengths, including Friedel pairs, is demonstrated (the theoretical minimum case); this is of particular interest when considering overall beam time needs and is clearly a feasible experimental strategy, as shown here. PMID- 16702656 TI - X-ray Holographic Microscopy of Biological Specimens with an Electronic Zooming Tube. AB - X-ray holographic microscopy of biological specimens has been performed using in line holography with the following improvements: (i) an electronic zooming tube is utilized as a detector, and (ii) the object planes are restricted by a metal frame which is not transparent to X-rays at the wavelengths used in the present experiments. The latter modification is useful in computer reconstruction and serves as a reference area where there are no photons in the reconstructed images. This system can minimize the inclusion of errors when recording and reading out holograms compared with methods using an X-ray resist as a detector, or optical reconstruction methods. Two-dimensional images of human cells have been successfully obtained with the present system. Comparison of these images with optical images revealed a good coincidence in structure outlines but not in internal structure images. Holographic observation of HeLa cells at different wavelengths showed a wavelength dependence of the reconstructed images. Since the electronic zooming tube is applicable to a wide range of X-ray wavelengths and has a promising potential to improve its resolution, these results strongly suggest that the present system is a good candidate for the development and establishment of an X-ray holographic microscopy system. PMID- 16702658 TI - Analysis of figure-8-undulator radiation. AB - The figure-8 undulator is a novel insertion device which has recently been proposed in order to obtain linearly polarized radiation with low on-axis power density. Since the trajectory is different from any other insertion device developed or proposed so far, the features of figure-8-undulator radiation are also considered to be different. In this paper, various features of figure-8 undulator radiation are investigated to clarify differences between the figure-8 undulator and other ordinary insertion devices. PMID- 16702659 TI - Development of a two-dimensionally tunable focusing monochromator for protein crystallography at high-energy undulator beamlines. AB - A two-dimensionally tunable focusing monochromator has been developed for protein crystallography at high-energy undulator beamlines of third-generation synchrotron radiation facilities. This monochromator consists of a silicon wafer fabricated with an oblique-cut angle between the Bragg net plane and the crystal surface, and adhered onto a table-like copper block. The radii of curvatures are altered independently in two directions by expanding the spaces between the table legs. The versatilities of the meridional and sagittal curvatures were confirmed by X-ray experiments and three-dimensional shape measurements, respectively. The two-dimensional focusing ability was demonstrated using high-energy X-rays of 37.7 keV emitted from a bending-magnet source at the Photon Factory. A quasi isotropic profile of converged X-rays was achieved near the focal position. The apparent gain of photon flux was 21. As a result of these excellent monochromator characteristics, a diffraction pattern of a hen egg-white lysozyme crystal was successfully obtained using high-energy X-rays. PMID- 16702660 TI - X-ray Monochromator with 2 x 10(8) Energy Resolution. AB - An X-ray beam of 3 x 10(7) photons s(-1) with 2 x 10(8) relative energy resolution has been obtained at a third-generation synchrotron undulator X-ray source using the (13 13 13) Bragg reflection from a silicon perfect crystal. The production of these 25.70 keV X-rays with 450 +/- 50 mueV bandpass opens up new possibilities in X-ray optics and spectroscopies. PMID- 16702662 TI - A new powder diffractometer for synchrotron radiation with a multiple-detector system. AB - A new powder diffractometer for synchrotron radiation with six detector arms has been constructed. Five detector arms are attached radially at intervals of 25 degrees to the 2theta axis and form a multiple-detector system. Five scintillation counters coupled with flat Ge(111) crystal analyzers on the respective arms can simultaneously record the whole powder pattern divided into five segments, each with an equal 2theta span. The optics design is based on flat specimen reflection geometry using a parallel beam. The intensity data are collected using a 2theta step-scan technique in asymmetric diffraction at a fixed incident angle. A sixth multi-purpose detector arm can be used in the conventional single-arm scan mode. It can be equipped with various kinds of analyzers such as long horizontal parallel slits, a fiat or channel-cut crystal analyzer, a receiving slit and a solid-state detector. Test operations of the multiple-detector system, conducted at the Photon Factory in Tsukuba, recorded a full width at half maximum of 0.022 degrees and a peak maximum intensity of more than 40000 counts s(-1) for the (111) reflection from Si powder. The whole powder pattern of Mg(2)SiO(4) over a 2theta range of 130 degrees could be step-scanned at a step interval of 0.004 degrees (2theta) in just 4 h. Results of whole-powder pattern decomposition and Rietveld refinement of the Mg(2)SiO(4) pattern are given. PMID- 16702661 TI - Feasibility and Realization of Single-Pulse Laue Diffraction on Macromolecular Crystals at ESRF. AB - Laue diffraction patterns with an exposure time of ca 60 ps have been acquired at the European Synchrotron Radiation Facility (ESRF) on protein crystals by using the single-bunch mode of the storage ring. A 10 ns laser pulse initiating photodissociation was synchronized with the X-ray pulse. The potential for a quantitative detection of conformational changes in proteins on the nanosecond timescale with this technique is demonstrated using the example of carbonmonoxymyoglobin, from simulations and real data. The instrumental aspects of the experiment (highly intense X-ray beam, fast shutter system, Laue camera, detector, laser apparatus and synchronization technique) are emphasized. PMID- 16702663 TI - Time-Resolved Simultaneous SAXS/WAXS of the Drawing of Polyethylene at the Daresbury SRS. AB - A system has been developed which represents a significant advance in the quality and extent of small- and wide-angle X-ray scattering data (SAXS and WAXS) that can be recorded simultaneously with strain data during the drawing and annealing of polymer materials. WAXS data are recorded using a Photonic Science charge coupled-device area detector and SAXS data using a gas-filled multiwire area detector. Strain data, for the region of the specimen from which the SAXS/WAXS data are collected, are calculated from an accurately synchronized continuously recorded video image of the specimen. The system allows X-ray and video image data to be collected as a series of frames with essentially no ;dead-time' between frames. The data are fully two-dimensional and can be collected for a wide range of d spacings. The use of this system to investigate the stress induced orientation and phase changes during the drawing of a range of grades of commercially available polyethylene is described. PMID- 16702664 TI - New commercial products. PMID- 16702665 TI - Phasing multi-segment undulators. AB - An important issue in the manufacture of multi-segment undulators as a source of synchrotron radiation or as a free-electron laser (FEL) is the phasing between successive segments. The state of the art is briefly reviewed, after which a novel pure permanent magnet phasing section that is passive and does not require any current is presented. The phasing section allows the introduction of a 6 mm longitudinal gap between each segment, resulting in complete mechanical independence and reduced magnetic interaction between segments. The tolerance of the longitudinal positioning of one segment with respect to the next is found to be 2.8 times lower than that of conventional phasing. The spectrum at all gaps and useful harmonics is almost unchanged when compared with a single-segment undulator of the same total length. PMID- 16702666 TI - Low-angle synchrotron radiation diffraction with glass-capillary optics. AB - The use of borosilicate-glass-capillary optics at a chosen wavelength for low scattering has been explored using an undulator beam at the ESRF. With a 2.3 mum beam at 0.092 nm wavelength, a silver behenate powder sample was scanned in two dimensions with a 2 mum step width. Scattering from single crystallites with d(001) = 5.83 nm could be observed. The limit for observation, at low angles, was ca s approximately 0.1 nm(-1) (s = 1/d for d approximately 10 nm). PMID- 16702667 TI - A foil-mask spectrometer for laue pattern imaging: simultaneous position, intensity and energy. AB - An X-ray spectrometer for simultaneous position, intensity and energy determinations suitable for Laue diffraction applications is described. The foil mask spectrometer consists of a series of metal foils of varying composition and thickness which are used to modulate the energy distribution of an incident X-ray source. Three modes of operation are described: a high-resolution spectrometer for measurement of nearly monochromatic X-rays, an intensity discriminator for partitioning the intensity from a small number of spatially overlapped monochromatic X-ray sources, and a low-resolution spectrometer for polychromatic X-rays with broad spectral features. The first mode of operation is designed to allow the energy of monochromatic Laue reflections to be measured with a resolution suitable for determination of unit-cell parameters. The second mode of operation is designed to allow the intensity of each component in a spatial region containing overlapping orders or spatially overlapped reflections to be discriminated for use in refinements or space-group assignment. The third mode of operation is described for completeness. The theory behind each mode of operation is described. The energy resolution of the spectrometer improves with the square root of the intensity of the incident beam. It also increases linearly with the change in energy with respect to transmission efficiency of a particular foil. In theory, the resolution of the spectrometer can readily exceed 50 eV over a wide range of energies depending on the foils used and the incident X-ray photon flux. Determinations of the energies of Mo Kalpha and Cu Kalpha radiation using a first generation ten-foil spectrometer gave values of 17.5 +/- 0.1 and 8.08 +/- 0.05 keV, respectively. Treatment of random error shows good correspondence with a Poisson model. The use of this spectrometer is demonstrated using a sample of tetraphenylphosphonium tetrachlorooxomolybdenum(V). Comparison of predicted and observed energies shows good agreement over a wide range of energies. The ratio of predicted to measured energy for the first 50 measurements was 0.9918+/ 0.0344. Up to three components of a position having harmonic overlap were separated. This work demonstrates the feasibility of using Laue diffraction to completely determine the crystal structure of a molecule without recourse to monochromatic methods. PMID- 16702669 TI - Analysis of EXAFS Data from Mixed-Shell Systems. AB - A new method for analysis of EXAFS data from coordination shells containing two types of atoms distributed at two well defined distances is proposed. The method, which in effect isolates the individual contributions of the two subshells, can be viewed as a refinement of conventional techniques such as beat analysis and multi-shell least-squares fitting. No external information on the structure of any of the contributing subshells is required beyond the usual assumption of small or ;Gaussian' disorder. As much as fivefold reduction in the confidence limits of the coordination numbers in comparison with unrestricted multi-shell fits is demonstrated. The range of applicability and limitations of the method are discussed in detail. PMID- 16702668 TI - Observation and modelling of preferred orientation in two-dimensional powder patterns. AB - A novel approach to the detection and modelling of preferred orientation is presented, based on the interpretation of two-dimensional powder patterns. A simple graphical construction is introduced to aid interpretation, and the application of this construction to some standard diffraction geometries is discussed. It is also shown in outline how a standard preferred-orientation model can be adapted to describe two-dimensional data. PMID- 16702670 TI - Characterization of Static Disorder by Cumulant Analysis of EXAFS: an Investigation on a Two-Gaussian Distribution. AB - The EXAFS of a two-Gaussian distribution is simulated. It is shown that the cumulant analysis of an EXAFS signal in the case of static disorder allows the reconstruction of the corresponding asymmetric interatomic distance distribution by the splice method. In addition, the relationships between the parameters of two Gaussians and the leading cumulants of their superposition are derived. The possibility of determining the parameters of the two Gaussians by analytical means is investigated. PMID- 16702671 TI - High-Spatial-Resolution Medical-Imaging System Using a HARPICON Camera Coupled with a Fluorescent Screen. AB - A high-sensitivity HARPICON camera was developed for medical X-ray imaging using a fluorescent screen. It is an avalanche-multiplication-type image pick-up tube and is 32 times more sensitive than conventional tubes. The camera also has a wider dynamic range than conventional medical-imaging cameras because a maximum output signal current of 2.3 muA is obtained and, in high-illumination-intensity regions, photocurrent is not proportional to illumination intensity. The fluorescent screen is an intensifying screen of the type used for radiographic screen-film combinations in medical examination. An X-ray image on the screen is focused on the photoconductive layer of the pick-up tube using a coupling lens with f/0.65. Experiments were performed using monochromated X-rays at the Photon Factory. An image of a spatial resolution test chart was taken in a 525 scanning line mode of the camera. The chart pattern of 5 line-pairs mm(-1 )(spatial resolution of 100 mum) was observed at an X-ray input field of 50 x 50 mm. Real time digital images of the heart of a 12 kg dog were obtained at a frame rate of 60 images s(-1) after injection of a contrast medium into an artery. The images were stored in digital format at 512 x 480 pixels with 12 bits pixel(-1). High spatial-resolution and high-contrast images of coronary arteries were obtained in aortography using X-rays with energy above that of the iodine K edge; the image quality was comparable with that of conventional selective coronary angiography. PMID- 16702672 TI - X-ray Optics of a Dynamical Sagittal-Focusing Monochromator on the GILDA Beamline at the ESRF. AB - The performance of a dynamical sagittal-focusing monochromator for hard X-rays is described. It consists of a flat first crystal and a diamond-shaped ribbed second crystal which is clamped by its central rib and dynamically bent by applying a force on its two apices. The system has proved to perform very well on the GILDA beamline at the ESRF. The horizontal acceptance varies with energy and with focusing geometry as predicted theoretically; the total available horizontal fan of radiation (3.6 mrad) is in fact collected in the 1:3 geometry. The system is routinely run in a dynamical focusing mode for XAFS spectroscopy in the energy range 5-30 keV with Si(311) crystals, with a constant spot size FWHM ~1 mm on the sample and without degradation of energy resolution or reproducibility. Using simple geometrical considerations we calculate the variations of the horizontal profile of the reflected beam during rocking-curve scans in different focusing geometries and find them in agreement with observed ones. Not only is this is a practical aid in alignment but it illustrates the X-ray optics of sagittal focusing in an elegant way. PMID- 16702673 TI - Wide dynamic range germanium detector for perturbation crystallography. AB - An IR detector based on a cooled germanium photodetector has been tested for applications in X-ray diffraction. The detector can be used simultaneously in photon-counting mode and current mode giving a dynamic range from < 1 to 10(9) photons s(-1). Since germanium is used as the photodetector, its efficiency at energies above 25 keV is much better than the silicon equivalents. The detector proved to be highly linear both in the low-flux region (< 10(5) photons s(-1)) where photon counting is used and in the high-flux region (> 10(5) photons s(-1)) where the detector is run in current mode. The response time of the detector is of the order of 1 mus, making it suitable for studies in perturbation crystallography, especially when coupled to a lock-in amplifier. As an example, the shift of a reflection of LiNbO(3) induced by an external electric field was determined with the germanium detector and lock-in amplifier. PMID- 16702674 TI - Application of a CdTe Solid-State Detector to Polarization-Dependent Total Reflection Fluorescence XAFS Measurements. AB - A CdTe solid-state detector was applied to the measurement of polarization dependent total-reflection fluorescence XAFS spectra. The data revealed that the detector has good sensitivity, and this, together with its compact size, make it appropriate for in-situ measurements and removal of X-ray Bragg diffraction. The detector efficiently recorded the high-energy K-edge XAFS spectra for molybdenum oxides supported on TiO(2) (110). PMID- 16702675 TI - Applications of synchrotron radiation to defect characterization and pendellosung fringe-spacing measurement in a natural diamond. AB - A small polished rectangular block of natural diamond unusually free from impurity zoning was examined by birefringence and synchrotron X-ray topography to map internal defects and assess effects of some accidental mechanical damage. ;Orthogonal-view' projection topographs facilitated comparisons between optical and X-ray images. Speedy and precise positioning of crystal sections cut in synchrotron X-ray section topographic studies was aided by radiographic and fluoroscopic procedures, which are described. Several X-ray reflection and refraction phenomena are illustrated, including high-contrast refraction images of crystal edges, chipping and crack outcrops. It was demonstrated that by combining optical and X-ray goniometry it was possible to record Pendellosung fringe systems employing precisely selected X-ray wavelengths in well defined crystal volumes. Valuable gain in fringe visibility resulted from the use of pure sigma-mode polarized synchrotron radiation. Absolute values of the diamond 004 structure factor were derived from five fringe systems measured, the findings (in upward order) being 11.7, 11.7, 12.0, 12.5, 12.6. Their scatter greatly exceeds the uncertainty in the measurement procedure (believed to be within +/-1%) and is attributed to different strain distributions within the five volumes sampled. PMID- 16702676 TI - New features of dislocation images in third-generation synchrotron radiation topographs. AB - Some aspects of the dislocation contrast observed at third-generation synchrotron radiation set-ups are presented. They can be explained by taking into account angular deviation effects on the beam propagation, which are visible because of the ;almost plane-wave' character of these sources. In particular, we show how the evolution of the direct image width of a dislocation as a function of the sample-to-film distance can allow a complete determination of the Burgers vector, i.e. in sign and modulus. In addition, experimental results obtained in monochromatic beam topography are compared with simulated images calculated assuming plane-wave illumination and are demonstrated to show a satisfactory agreement. The utility of the weak-beam technique in enhancing the spatial resolution is demonstrated and a criterion for the selection of experimental conditions depending upon the required spatial resolution, signal-to-noise ratio and exposure time is presented. PMID- 16702677 TI - State-of-the-Art Analysis of Whole X-ray Absorption Spectra. AB - Fitting an entire X-ray spectrum rather than its components, EXAFS and XANES, has been an aim of the practitioners of these techniques. Recent developments have made the calculations of both the scattering and atomic components practicable. We present the analysis of four representative model compounds using the EXCURVE package, which was modified to undertake this. The details of these modifications are also given. A comparison of matrix-inversion and finite-path-sum methods is made which shows that the latter method is more promising for fitting the edge region. A number of enhancements are required before this approach can be used for accurate structure determination. These include improvement in atomic contribution, better potentials/phase shifts, and the ability to calculate and refine multiple-scattering terms to at least fifth order. PMID- 16702678 TI - A monochromator for scanning X-ray microscopy beamlines at third-generation synchrotron light sources. AB - A concept for a plane-grating monochromator for use at scanning X-ray microscopy beamlines at third-generation synchrotron light sources is presented. The design of the monochromator is optimized for a scanning transmission X-ray microscopy beamline at BESSY II. Ray-tracing calculations are presented which include geometric aberrations of the optics used in the beamline. PMID- 16702679 TI - Production of intense low-energy positron beams with synchrotron radiation. AB - A low-energy positron beam is a unique probe of Fermi surfaces, defects, surfaces and interfaces. In high-energy electron and positron storage rings (E > 6 GeV) it is possible to generate intense synchrotron radiation with 1-3 MeV photons by installing a high-field superconducting wiggler. The strength of the wiggler should be ~8-12 T. High-energy photons are emitted from the wiggler and converted to low-energy positrons by using a suitable target-moderator system. For an 8 GeV electron storage ring at a beam current of 100 mA, final yields are estimated to be ~10(10)-10(12) (slow-e(+) s(-1)) with the size of positron source ~10(2)-10(3) cm(2). The possibility of increasing the brightness of the low-energy positron beam is discussed. Advantages of using synchrotron radiation for producing positrons are pointed out. The effect of a superconducting wiggler on the stored electron beam is also discussed. PMID- 16702680 TI - Phase shift and radiation spectrum of a straight series of undulators. AB - In constructing a very long undulator it is convenient to separate the undulator into a number of sections with a constant gap distance between the sections. The properties of the radiation spectrum expected in such an undulator were investigated assuming ideal short undulators for each section. It is shown that the peak height and width of the spectrum do not deteriorate if the radiation phases of the sections are matched to each other, even with a large gap distance. This can be realized by adjusting the field strength of the three-pole wigglers installed in each gap. PMID- 16702681 TI - PIN Silicon Diodes as EXAFS Signal Detectors. AB - The properties of PIN silicon diodes as X-ray detectors for EXAFS measurements with synchrotron radiation have been investigated. Electronic stability, linearity and noise current have been analyzed. The effects of diffraction peaks resulting from the crystalline nature of the diodes have been minimized by mounting the diodes on a simple device that continuously changes its orientation by a few degrees with respect to the X-ray beam. An accurate comparison between EXAFS signals monitored by ionization chambers and PIN photodiodes is presented. It is shown that good-quality EXAFS measurements with PIN photodiodes are possible if diffraction effects are eliminated. PMID- 16702682 TI - High-Pressure Cell for the Study of In-Situ Hydrates Using Energy-Dispersive X ray Diffraction. AB - A high-pressure low-temperature cell has been developed for in-situ diffraction studies of carbon dioxide and propane gas hydrate crystallization. The design and implementation of the cell, which can operate up to 3.5 MPa and down to 253 K, are described. Using synchrotron energy-dispersive X-ray diffraction, the first growth of the hydrate crystals from solution has been successfully observed. The lattice parameters of the hydrate crystals were found to be 11.927 (2) and 17.196 (2) A, respectively. PMID- 16702683 TI - Dynamic Small-Angle X-ray Scattering System using an Imaging Plate. AB - A synchrotron-radiation dynamic small-angle X-ray scattering (Synchrotron Radiation-DSAXS) system using an imaging plate has been developed at the Photon Factory, National Laboratory for High Energy Physics, Japan, in collaboration with the Department of Polymer Chemistry, Kyoto University, Japan, for studying the dynamic response of mesoscopic structures in polymers to applied mechanical stimuli. Small-angle X-ray scattering patterns of 100 x 100 mm(2) have been successively recorded on a 400 x 200 mm(2) stage-mounted imaging plate with a minimum time interval of 0.5 s. X-ray data are simultaneously obtained with stress and strain measurements on specimens subjected to a mechanical deformation. The performance of the DSAXS system is demonstrated along with some experimental results concerning the dynamic deformation of a b.c.c. lattice with paracrystalline distortion in a co-polymer having spherical microdomains under a large oscillatory shear deformation. PMID- 16702684 TI - Soft X-ray Emission Spectroscopy at ESRF Beamline 26 Based on a Helical Undulator. AB - A new soft X-ray spectrograph for X-ray emission spectroscopy excited by synchrotron radiation is presented. The apparatus is now installed on beamline 26 at the ESRF in Grenoble. A brief description of the beamline is given and then several components of the spectrograph are covered in more detail. Results of experiments performed both with direct non-monochromated undulator radiation and with monochromated radiation are reported. PMID- 16702686 TI - New Editor-in-Chief of IUCr journals. PMID- 16702685 TI - Photoelectron holography of the si(001) surface. AB - Three-dimensional images of the near-surface atom arrangement were calculated from two-dimensional photoelectron diffraction data by several imaging algorithms: (i) a basic method with a Fourier transformation at one kinetic energy over k space, considering the phase factor due to the path-length difference; (ii) energy summation of the above results; (iii) Fourier transformation within small k-space windows; and (iv) their combinations. Atomic images produced by these methods from the experimental Si 2p photoelectron diffraction patterns of an Si(001) surface are compared with the crystal geometry. The results show that the energy-summed small-window method, called SWEEP, gives the best images. PMID- 16702687 TI - JSR: Required Reading. PMID- 16702688 TI - Theo thole, 1950-1996. PMID- 16702689 TI - Recording diffraction data for structure determination for very small crystals. AB - An account is given of experiences in recording diffraction data with synchrotron radiation to determine structures for very small crystals, too small for use with conventional X-ray sources and diffractometers. The effect of crystal composition as well as size on the relative intensity of diffraction patterns of different crystals is noted. Crystal mounting is briefly described. Experimental methods are outlined including detectors and other instrumentation, and a range of examples is given; these include the use of both monochromatic area-detector systems for intensity measurement and of the white-beam Laue method. Choice of the shortest wavelength available with adequate intensity is recommended. The examples include organic, organometallic and aluminophosphate compounds; in all cases structure determination was important in relation to chemical research projects - they were not chosen as ;test' crystals. Comparison is made of the quality of the structure refinements achieved with those from synchrotron radiation powder diffraction - the alternative method when good-sized crystals are not available. Commonly it is found that when good-quality large crystals of a substance cannot be grown, the small crystals are poor in quality with substantial mosaic spread; the relationship between mosaic spread, structure, morphology and crystal growth is explored. PMID- 16702690 TI - Studies of the Structure of Matter with Photons from an X-ray Free-Electron Laser. AB - X-ray free-electron lasers offer the potential for fourth-generation ultra-high peak power and coherence X-ray beams in the hard X-ray (8 keV) spectral range. A critical review of three areas of potential application of such sources is presented. Owing to the enormous electric field strength at the focus of such beams, it is shown that matter would very rapidly form a plasma for focal spots of submicrometer dimensions. Thus, hologram formation would only be feasible for crystalline samples with dimensions of a few micrometers or greater. A new version of X-ray holography using interference between a sample and a reference two-dimensional crystal is proposed. Because of the very high peak intensity of the free-electron-laser pulses, a natural application is to time-resolved multibunch ;dynamic X-ray scattering' measurements. It is shown that this could provide information on dynamical processes in condensed matter complementary to that obtained using slow neutron beams. Finally, optical laser-induced pump-probe type experiments are reviewed. There is the potential for extending the study of the charge distribution of electronic excited states to those with lifetimes in the submillisecond time range. PMID- 16702691 TI - A technique for generating potent positron beams. AB - This paper reports on a novel scheme that has the potential to generate intense positron beams. It is based on the modified betatron accelerator, a compact high current device that has been developed in the last few years. Briefly, the proposed accelerator consists of two modified betatron accelerators that are stacked together and share the same core. The electrons are accelerated in the upper torus during the first half of the flux waveform when the time rate of the magnetic flux (varphi) is positive. After completion of the acceleration, the electrons are extracted and guided to a high-Z target producing a positron beam that is accelerated in the lower torus during the second half of the waveform when dvarphi/dt is negative. PMID- 16702692 TI - Design of an X-ray Phase-Plate Analyzer to Measure the Circular Polarization Rate of a Helical Undulator Source. AB - A quarter-wave plate made of a ca 16 mum-thick silicon single-crystal was used at energies as low as 2.8 keV to convert circularly polarized photons into linearly polarized photons. Coupled to a linear polarimeter, this quarter-wave plate enabled the characterization of the circular polarization rate of the radiation emitted by one of the ESRF helical undulators, Helios-I. The measured value (ca 97%) is in good agreement with theoretical predictions. Special attention was paid to the alignment procedures of all relevant optical components of the beamline. PMID- 16702693 TI - Design of a Holographically Recorded Plane Grating with a Varied Line Spacing for a Soft X-ray Grazing-Incidence Monochromator. AB - A new design concept is presented for a plane grating with a varied line spacing for the Monk-Gillieson mounting monochromator. A light path function including both a spherical mirror and a varied-line-spacing grating is defined to optimize groove parameters. Aspheric wavefront recording optics are utilized to fabricate a grating holographically. Ray-tracing results show that the varied-line-spacing grating eliminates aberrations significantly and affords a high resolving power as a total optical system of a soft X-ray grazing-incidence monochromator. The effects of errors in recording parameters and in the radius of the spherical mirror are described, and possible ways to compensate for these errors are discussed. PMID- 16702695 TI - PXGEN: a General-Purpose Graphical User Interface for Protein Crystallography Experimental Control and Data Acquisition. AB - PXGEN is a general-purpose graphical user interface for experimental set-up and control of protein crystallography data collection. PXGEN is not linked intrinsically to any software package or proprietary hardware and should be transportable to other experimental facilities. The experimental techniques supported are single-wavelength data collection and multiwavelength anomalous dispersion. The graphical user interface runs on a UNIX-based workstation exploiting the host's power to manage multiple programs. PXGEN provides a mechanism for making data collection much easier and less error-prone. The design and implementation of PXGEN are described, which is now installed on protein crystallography beamlines 9.5 and 7.2 of the Synchrotron Radiation Source at Daresbury Laboratory. PMID- 16702694 TI - Protein Crystal Diffraction Patterns Using a Capillary-Focused Synchrotron X-ray Beam. AB - A paraboloidally tapered glass monocapillary was used to focus an 8 keV monochromated synchrotron bending-magnet X-ray beam into a 40 (+/-5) mum focal spot located 45 (+/-5) mm from the exit of the capillary. This focal spot had a measured intensity gain of 120 (+/-10) times the intensity present in an equivalent cross section of the unfocused beam from the monochromator. This focused beam was used to obtain oscillation diffraction patterns on image plates from a hen egg-white lysozyme protein crystal in two distinct geometries: one with the specimen crystal at the capillary exit and the other with the crystal at the beam focus. In the first geometry, focused Bragg reflections were observed at the focal plane. In the second geometry, diverging Bragg reflections of high intensity from a small crystal volume were observed. Image-plate diffraction patterns for these two geometries were compared with exposures with equivalent integrated diffracted intensities obtained using a 100 x 100 mum unfocused X-ray beam with the same crystal. The use of the focused beam resulted in a reduction in the exposure time required to produce equivalent patterns by a factor of between 70 and 100. PMID- 16702696 TI - In Situ Single-Crystal X-ray Diffraction Study of Crystallization Kinetics in Clathrasil Dodecasil-3C. AB - The formation of single crystals of the clathrasil dodecasil-3C from a solvothermal synthesis has been followed by in situ diffraction techniques using synchrotron radiation and an image-plate area detector. The high intensity of the X-ray beam, coupled with the ability to record time-resolved two-dimensional data using the image plates, allowed the crystallization kinetics to be studied and rate expressions to be fitted to the crystallization curves. PMID- 16702697 TI - Small-Angle X-ray Diffraction of Muscle Using Undulator Radiation from the Tristan Main Ring at KEK. AB - Time-resolved X-ray diffraction of muscle has demanded ever-increasing flux into small sample volumes with low beam divergence. Results are reported of static and time-resolved small-angle X-ray diffraction studies on muscle fibers using a hard X-ray undulator installed in the Tristan main ring at KEK, Tsukuba, Japan, as an innovative source of synchrotron radiation more intense and better collimated than that available with the Photon Factory bending-magnet beamline. Static studies used the low divergence of the source to obtain detailed high-quality diffraction patterns of stable muscle states. The diffraction patterns from live skeletal muscles showed the numerous (over 100) meridional reflections. The well collimated beam from the undulator made it possible to clearly resolve, with an angular resolution of ca 700 nm, the closely spaced diffraction peaks arising from the two halves of the thick filaments centred on the M lines in a sarcomere, in addition, the diffraction peaks from the thin filaments on opposite sides of the Z bands could be resolved with an angular resolution of ca 1000 nm. The detailed structure of the meridional pattern defines the nature of the molecular packing in the thick and thin filaments. Time-resolved experiments using a focusing mirror aimed to prove cross-bridge states in striated muscle fibers by collecting X-ray diffraction data at a 0.185 ms time resolution from sinusoidally oscillating chemically skinned rabbit muscle fibers during active contraction and in rigor. When sinusoidal length changes at 500 Hz with a peak-to-peak amplitude of 0.6% of the muscle length were applied to a small fiber bundle, the tension showed a simple elastic response during the length oscillation. In the active muscle the intensity of the 14.5 nm myosin-based meridional reflection changed out of phase with the tension change during the oscillating length change. In contrast, in the rigor muscle it occurred in phase with the tension change. The high time-resolved experiments provide an insight into the coupling between conformational changes and force generation of the actomyosin cross-bridges. These studies provide a preview of the expected gains for muscle studies from the more widespread use of undulator radiation at third-generation synchrotron sources. PMID- 16702698 TI - Reticulography: a simple and sensitive technique for mapping misorientations in single crystals. AB - Interposition of a fine-scale X-ray absorbing mesh between a Laue-diffracting crystal specimen and the photographic plate recording its topographic image splits the diffracted beam into an array of individually identifiable microbeam elements. Direction differences between the microbeams in the array, which are twice the orientation differences between the crystal elements reflecting them, are measured by recording the array at two or more mesh-to-photoplate distances. Maps of misorientation vectors over the crystal lattice planes under examination can be derived from these array images by visual or digital electronic metrological procedures. Applications to two specimens widely different in diffracting properties are described. Angular size of the X-ray source is the principal instrumental factor setting misorientation detection limits, and was less than 1 arc second in this work. PMID- 16702699 TI - First observation of inelastic X-ray scattering from condensed (4)He using high energy resolution. AB - Recently, the suitability of inelastic X-ray scattering for the investigation of solid and fluid (4)He has been demonstrated. For the test experiments an energy resolution of the order of 10-15 meV was used at the backscattering spectrometer INELAX at the storage ring DORIS of DESY, Hamburg. Lattice excitations were observed for momentum transfers along the c axis of h.c.p. helium crystals which were grown in situ at pressures of 54-63 MPa and at temperatures of 4.2-6.4 K. At 10 K above the melting point, energy-loss signals could also be detected from the liquid helium at equivalent momentum transfers. PMID- 16702700 TI - Prices of IUCr journals. PMID- 16702701 TI - Medicinal mushrooms and cancer therapy: translating a traditional practice into Western medicine. AB - Modern medical practice relies heavily on the use of highly purified pharmaceutical compounds whose purity can be easily assessed and whose pharmaceutical activity and toxicity show clear structure-function relationships. In contrast, many herbal medicines contain mixtures of natural compounds that have not undergone detailed chemical analyses and whose mechanism of action is not known. Traditional folk medicine and ethno-pharmacology coupled to bioprospecting have been an important source of many anticancer agents as well as other medicines. With the current decline in the number of new molecular entities from the pharmaceutical industry, novel anticancer agents are being sought from traditional medicine. As the example of medicinal mushrooms demonstrates, however, translating traditional Eastern practices into acceptable evidence-based Western therapies is difficult. Different manufacturing standards, criteria of purity, and under-powered clinical trials make assessment of efficacy and toxicity by Western standards of clinical evidence difficult. Purified bioactive compounds derived from medicinal mushrooms are a potentially important new source of anticancer agents; their assimilation into Western drug discovery programs and clinical trials also provides a framework for the study and use of other traditional medicines. PMID- 16702702 TI - Inflammatory bowel disease: maladaptation of the vigilant genotype in a hyper clean world? AB - Children born of Asian Indian parents who are raised in environmentally hygienic Western societies appear to be highly prone to two diseases, ulcerative colitis and Crohn's disease, collectively known as inflammatory bowel disease or IBD. These ethnically Indian children are similar to an inbred mouse strain, NOD/Lt. Mice of this strain remain diabetes-free when raised in standard mouse colonies, but develop an autoimmune diabetes at high rates when kept in pathogen-free environments. I propose that certain human habitats have, over eons, selected for "vigilant genotypes," wherein combinations of alleles at critical loci result in aggressive immune responses to pathogens. This genetic configuration is adaptive in the selective environment but maladaptive in more hygienic environments, resulting in dysregulated immune effectors. One manifestation of such dysregulation is organ-specific autoimmunity, such as IBD. PMID- 16702703 TI - Placebo, meaning, and health. AB - Placebos are boon and bane to medical theory and clinical practice. On the one hand, randomized controlled trials employ concealed allocations of placebo to control for effects not due to specific pharmacological mechanisms. As a result, nearly all of evidence-based medicine derives from principles and practices based on placebo. On the other hand, medical researchers and physicians have tended to ignore, minimize, or deride placebos and placebo effects, perhaps due to values emphasizing scientific understanding of mechanistic pathways. We argue that intention, expectation, culture, and meaning are central to placebo-effect phenomena and are substantive determinants of health. We introduce three dualities that are integral to placebo/meaning phenomena: body-mind, subconscious conscious, and passive-active. These placebo-related dualities should be acknowledged, explored with research, and incorporated in theory. While we view consideration of placebo and meaning effects as essential to any adequate understanding of human health, we feel that lessons from this area of inquiry may already provide practical tools for astute clinicians. Toward this end, we list eight specific clinical actions: speak positively about treatments, provide encouragement, develop trust, provide reassurance, support relationships, respect uniqueness, explore values, and create ceremony. These clinical actions can empower patients to seek greater health and may provide a healthful sense of being cared for. PMID- 16702704 TI - The prescriber as moralist: values in the antidepressant debate. AB - The past decade has seen a growing debate about the expanding use of psychotropic medications. Of particular concern are current antidepressants, as well as hypothetical "mood brighteners" that could modify affect and behavior in people heretofore classified as being within the normal range. This paper argues that objections to such pharmacologic applications are based on appeals to cultural values--authenticity, diversity, inwardness, and stoicism among others--that are viewed as being under increasing threat in contemporary American culture. Critiques of mood brighteners, like critiques of technological, consumer-driven culture, repudiate the ideals of shallow satisfaction and of the self as commodity. The decision to prescribe or not to prescribe a psychotropic medication in any given case is based on cultural values as well as clinical judgment. PMID- 16702705 TI - Dr. Arthur Conan Doyle and the case of congenital syphilis. AB - In 1894, Dr. Arthur Conan Doyle wrote "The Third Generation," a short story involving the transmission of congenital syphilis from generation to generation. Analysts of his writings have interpreted the pathogenetic mechanism involved in modern terms: infection of mother by father and then transplacental infection of the fetus. However, a review of the contemporary literature and the history of the concepts of congenital and "hereditary" syphilis demonstrates that the late 19th-century understanding of the process involved a Lamarckian transmission of paternal infection, via the sperm at the moment of conception. It was undoubtedly this concept that Doyle learned in medical school in the late 1870s and that provided the background to his story. PMID- 16702706 TI - A clinical alternative to the public health approach to mental illness: a forgotten social experiment. AB - In the last half of the 20th century, the community mental health movement, based on a public health model, came to dominate patterns of care for mental patients. In the process, brutal deinstitutionalization of very ill patients took place, at least in the United States. These events were not inevitable. In 1949, the Menningers of Topeka, Kansas, began administering Topeka State Hospital, which was in deplorable condition. By concentrating expenditures on clinical personnel, the Menningers humanely deinstitutionalized many patients before chlorpromazine, before the entitlement programs of the U.S. federal government such as Medicaid (1965), and before the community psychiatry movement got under way. Topeka State Hospital furnished a model of mental health care that centered a whole system on a last-resort, large, specialized state mental hospital. This inadvertent social experiment suggests that a clinical approach to mental health care offers a hard headed alternative to present arrangements. PMID- 16702707 TI - Strong Inference: rationale or inspiration? AB - John Platt's article "Strong Inference" (1964) suggested a general and effective method of scientific investigation. It describes a disciplined strategy of falsification of multiple, clearly formulated hypotheses that is used more regularly in some scientific fields than in others. Platt urged that strong inference be more widely and more systematically applied, particularly in slower moving fields of science. The article has influenced integrative biological fields since its publication, ranging from ecology to psychology, and has had a substantial following in some of the social sciences. It has also evoked severe criticism for its idealization of certain fields as exemplars and for its imperfections in historiography and philosophy of science. I argue here that the article was more an inspirational tract than the development of a formal scientific methodology. Although both Platt's critics and his adherents appeared to take the article far too seriously, its influence has transcended its limitations. PMID- 16702708 TI - Investigating the Barons: narrative and nomenclature in Munchausen syndrome. AB - Recounting stories of the 18th-century Baron von Munchhausen, the unauthorized appropriation of his name into literature, and Richard Asher's subsequent medical use of the name Munchausen 150 years later, this article examines the narratives that are told about and through Munchausen syndrome that create meaning within medicine. By analyzing a half-century debate over the name of the illness, this article discusses how the medical literature invests meaning in names and how names create meaning through narrative with effects on the practice of medicine. PMID- 16702709 TI - Beyond the physician charter: reflections on medical professionalism. AB - The numerous challenges now facing the profession of medicine have led to an intense focus on professionalism by individual physicians and by their professional and academic organizations. In 2002, a distinguished group of leaders in internal medicine created the Physician Charter, which calls on physicians to reaffirm medical professionalism through commitment to three principles and 10 responsibilities. The Charter reflects a duty-based ethic that is chiefly concerned with physician competence. This article offers a critical analysis of the Physician Charter from the perspective of the traditional values of medicine as articulated in medical oaths and championed by leaders of past generations, exemplified by William Osler. The authors argue that medical professionalism should reflect the values of a virtue-based ethic that stresses compassion and beneficence, rather than the values of a duty-based ethic. The challenges that now confront the practice of medicine can be addressed successfully only to the extent that physicians promote virtue-ethics, act collectively in the public interest, and render service that clearly transcends their own self-interests. PMID- 16702710 TI - Medicine in the age of " Ulysses ": James Joyce's portrait of life, medicine, and disease on a Dublin day a century ago. AB - Over time, contemporary writing becomes part of the historical record. In medicine, it is an important learning tool, particularly for understanding the experience and context of disease and illness. Although a century has elapsed since the fictional events on a single day described in James Joyce's Ulysses, the work is still fresh with references and allusions to doctors, illnesses, and the human experience. Ulysses provides perspective on medical and social history and offers a biting commentary of continuing relevance to the doctor-patient relationship. PMID- 16702712 TI - Minority faculty recruitment, retention and advancement: applications of a resilience-based theoretical framework. PMID- 16702713 TI - Understanding the success of promotoras in increasing chronic diseases screening. AB - The purpose of this qualitative study was to elicit information on why a promotora (or, community health worker (CHW)) increased adherence to chronic disease screening among women along the U.S.-Mexico border. After completion of the intervention, women and clinic staff who participated in the promotora phase of a randomized, controlled study answered structured, open-ended questionnaires. Clinicians from two non-participating clinics were also interviewed. Content analysis found that the promotora's roles included health education and the facilitation of routine and follow-up care. Clients appreciated the promotora's socio-cultural characteristics, as well as her personal skills and qualities, and described her as a trained, natural helper whose personalized support removed barriers to health care and helped women to take care of themselves. Most clinicians recommended working with a CHW to increase adherence to chronic disease prevention practices. A CHW can play a crucial role on a health care team and interventions should tap into this resource. PMID- 16702714 TI - Health care utilization and risk behaviors among HIV positive minority drug users. AB - This study compared health care utilization and HIV-related risk behaviors between HIV-infected African American (n=123) and Hispanic (n=97) drug users recruited in New York City. African Americans were more likely to use crack, while Hispanics were more likely to use heroin and speedball. African Americans were more likely than Hispanics to report having traded sex for drugs or money. The two groups did not significantly differ in HIV care utilization (e.g., taking HIV medications). In multiple logistic regression analyses, for African Americans, taking HIV medications was significantly related to enrollment in HIV clinics and non-use of crack, while for Hispanics, being married and attending HIV support groups were significant factors. Drug treatment enrollment was significantly related to non-use of crack and injection drug use among African Americans, and a trend (p=.07) was found between injection drug use and drug treatment enrollment among Hispanics. The findings indicate the continuing need for harm reduction programs for HIV positive drug using populations. Programs for these populations should also take into account the different risk behaviors among different racial/ethnic groups, in order to tailor culturally sensitive programs for HIV care and intervention. PMID- 16702715 TI - The medically underserved: who is likely to exercise and why? AB - Adults who exercise regularly have better health, but only 15% of U.S. adults engage in regular exercise, with some social groups, such as people with lower incomes and women, having even lower rates. This study investigates the rate at which medically underserved patients receive exercise counseling from health care providers, characteristics of those who exercise, and barriers and motivations to exercise. The convenience sample was predominantly female and White and exclusively low-income and uninsured or underinsured. On average, participants were obese, by Federal Obesity Guidelines; 43% smoked. Although 60% of 126 patients reported that providers discussed exercise with them, the discussions had no relationship with patients' engagement in exercise. Women and those with lung problems, diabetes, or children in the home were less likely than others surveyed to exercise. The highest rated motivations included body image and health issues. The most important barriers were time, cost, and access to exercise facilities and equipment. In order for exercise counseling to be more effective, health care providers' interventions must consider patients' personal characteristics, health status, readiness to engage in an exercise program, and motivations and barriers to exercise. PMID- 16702716 TI - Cardiac risk factors, medicine usage, and hospital course in immigrants from the former Soviet Union. AB - Immigrants from the former Soviet Union have a higher prevalence of cardiac risk factors and more problems obtaining health care in the United States than American-born Caucasians. This study compared differences between patients of these two populations admitted for diagnosis of chest pain or shortness of breath. Immigrants from the former Soviet Union (who had been in the U.S. for an average of 20 years) had more cardiac risk factors than American-born Caucasians including more hypertension (81% vs. 50%, p=.002), positive family history (53% vs. 30%, p=.030), more previous heart attacks (45% vs. 20%, p=.012), more prior cardiac catheterizations (51% vs. 18%, p<.001) and coronary revascularization procedures (51% vs. 27%, p=.022), and higher systolic blood pressure (138+/-13 vs. 129+/-23 mmHg, p=.019) upon presentation to the hospital. Fifty-five percent of immigrant patients used foreign medications. Thus, there are major differences between immigrants from the former Soviet Union who are admitted to the cardiac units of an urban New York hospital and American-born Caucasians. Knowledge of these differences is important for caregivers. PMID- 16702717 TI - Implementing ADHD guidelines in primary care: does one size fit all? AB - To determine if the American Academy of Pediatrics Attention Deficit/Hyperactivity Disorder (ADHD) guidelines require tailoring for different settings, the researchers used a mixed-method research design to review an ADHD quality improvement effort in community clinics and private offices in San Diego County. Clinically, no differences were noted in rates of ADHD in the two settings. Children in community clinics (58.3%) were more likely to report public insurance (p<.001), diverse ethnic backgrounds (p=.003), low household incomes (p<.001), single parent households (p=.009), and to screen positive for Oppositional Defiant Disorder/Conduct Disorder (p=.027). They were also more likely to have experienced socio-environmental stressors (p<.001) including foster care, homelessness, parental drug use, and domestic violence. No differences were noted by treatment received at 12 months post-evaluation by office type. Open-ended interviews with clinicians confirmed these findings and revealed a need for tailoring of implementation strategies to more closely fit the needs of children and families cared for in public sector settings. PMID- 16702718 TI - Factors associated with chronic conditions among children in foster care. AB - The population of children in foster care is rapidly growing. Previous local and state-level analyses have measured the prevalence of chronic conditions among such children to be from 44%-82%. The study objective was to identify factors associated with chronic conditions among a nationally representative sample of children in foster care for one year. The authors analyzed data from The National Survey of Child and Adolescent Well-Being (NSCAW), Wave 1, the first national dataset of children in the child welfare system. In regression analysis, factors significantly associated with having a chronic condition included: child age under 2 years, caregiver race/ethnicity other than Hispanic, and relatively few household members. Discussion includes consideration of chronic conditions in this high-risk population. PMID- 16702719 TI - Provider recognition of psychosocial problems in low-income Latino children. AB - The purpose of this study was to determine the frequency of identification of psychosocial problems in Latino children by primary care providers (PCPs). This was a cross-sectional study of 269 low-socioeconomic status (SES), Latino children, ages 2-16 years, seen at a community clinic for well-child care. Primary care providers completed a World Health Organization checklist, which organized provider assessment of child psychosocial problems. Clinicians identified one or more psychosocial problems in 39.8% of children. Clinicians recognized only 20% of children with clinically significant aggression symptoms, 18% of children with clinically significant attention/hyperactivity symptoms, and none (0%) of the children with clinically significant anxiety/depression symptoms with clinical data from parent-completed child-behavior checklists used as the benchmark. Despite high rates of identification of psychosocial problems in a low SES, Latino population, PCPs still miss symptoms of mental health difficulties, especially anxiety and depression. Since symptoms of anxiety and depression are likely to be more pronounced in Latino populations, there is a continued need for improvement in detection of mental health concerns among these children. PMID- 16702720 TI - Child passenger safety behaviors in Latino communities. AB - Booster seats protect child occupants between 4 and 8 years of age. The objective of this study was to determine barriers and facilitators for booster seat use among Latino families. We conducted one-to-one elicitation interviews with 56 mothers and 35 fathers of booster-eligible Latino children in an urban county and a rural county in Washington State. Half of the parents did not consistently use booster seats. Interviews were recorded, transcribed, translated, and coded. Statements expressed by at least one-third of respondents were entered into explanatory models. Motivators for booster use were child safety and concern about getting a ticket. Facilitators for booster use included affordability, ease of use, and children liking the seat. Barriers were the belief that the child was too big/old, perceived child resistance, and cost. Rural parents preferred radio to television messages. Campaign messages highlighting the risks to child safety and the risk of a citation are likely to motivate booster seat use among Latino families. PMID- 16702721 TI - Access to health care for migrants returning to Mexico. AB - Continued migration from Mexico over the past several decades has created a large population of elderly Mexicans in the U.S. There is no system in Mexico for those Mexicans who would like to retire there to obtain health insurance during their retirement years. Using a nationally representative dataset of Mexican elders, we explore the current state of health insurance status for Mexican elders with a history of migration to the U.S. We find a robust negative association between years spent in the U.S. and the probability of being insured. Coordination between the U.S. and Mexico on policy options to insure Mexicans migrants may prove beneficial to the social security systems in both countries as well as to migrants themselves. PMID- 16702722 TI - Male Hispanic immigrants talk about family planning. AB - In order to improve delivery of family planning services by better understanding the views of people affected, researchers conducted four focus groups among Hispanic men who recently migrated to a Midwestern community. Participants expressed opinions about, attitudes towards, and knowledge of such matters as family planning, birth control information and access, the influence of religion on family planning and birth control decisions, general familial relations, the prevalence of infidelity, use of and access to the health care system, and cultural differences between the U.S. and their countries of origin. The men voiced opinions and attitudes supportive of family planning and appeared knowledgeable about various methods of birth control. Understanding the attitudes held by immigrant men is important for health care providers in designing successful service interventions for this rapidly growing segment of the population. PMID- 16702723 TI - Cognitive abilities of African American children with prenatal cocaine/polydrug exposure. AB - The aim of this study was to investigate the effects of prenatal cocaine/polydrug exposure on cognitive abilities of school-aged children. The study examined differences in the performance of low income prenatal cocaine/polydrug-exposed and non-exposed African American children (n=49) between 6 and 8 years of age, on the Wechsler Intelligence Scale for Children-Third Edition. Twenty-five children had a history of prenatal cocaine/polydrug exposure and 24 had no known history. The groups were matched for age, sex, and socioeconomic status. Test scores determined if drug exposure predicted global intelligence score and if there were significant differences between the two groups in specific areas of cognitive ability assessed. Drug exposure predicted IQ scores, as drug-exposed children performed significantly lower than their non-exposed counterparts on global intelligence and verbal comprehension, but comparably on the freedom from distractibility tasks. On individual cognitive tasks, the performances of cocaine/polydrug-exposed and non-exposed children were comparable on 8 of the 11 tasks assessed. The findings call into question earlier predictions about cocaine exposed children. PMID- 16702724 TI - Characteristics of six-month tobacco use outcomes of Black patients seeking smoking cessation intervention. AB - Although Blacks experience disproportionately greater morbidity and mortality attributable to smoking than other racially-classified social groups, few studies have examined the impact of clinical interventions for nicotine dependence within this population. The main objective of this study was to examine 6-month outcomes among 146 self-identified adult Black patients who received an individually tailored nicotine dependence intervention in an academic medical setting. Measures included a baseline demographic questionnaire and telephone follow-up to obtain self-reported 6-month tobacco use status. Univariate analysis was performed to assess the association of baseline patient characteristics with tobacco abstinence at 6 months following the clinic intervention. Of the 146 patients, 83% were seen in an outpatient clinic setting, while 17% were seen as inpatients in the hospital. At baseline, 53% reported smoking an average of 20 or more cigarettes per day, 32% were highly nicotine dependent, and 53% were in the preparation or action stage of change. Six months following the intervention, the 7-day point-prevalence tobacco abstinence rate was 43/146 (29%; 95% C.I. 22% to 37%). An individualized nicotine dependence intervention conducted in an academic medical setting yielded encouraging abstinence rates for Black smokers. PMID- 16702725 TI - Housing outcomes for hospitalized homeless veterans. AB - This study examines housing status at the time of hospital discharge in a national sample of 3,502 veterans who were homeless at admission to a Veterans Health Administration (VHA) medical center. A supplemental survey on homelessness was added to the annual VHA inpatient census over a 4-year period (1995 through 1998). Data from this survey were used to identify homeless veterans hospitalized on the final day of the fiscal year and to document their housing status at discharge. A signal detection technique, Receiver Operating Characteristic analysis, was used to identify predictors of housing status at discharge. At discharge, only 13% of the veterans in our analytic sample were literally homeless, 40% were doubled up, and 33% were transferred to another an institution; only 13% were living independently. Housing status at admission (specifically, staying with friends or family temporarily rather than literally homeless), treatment in a psychiatric or substance abuse program rather than a medical program, and a greater income level or access to VHA benefits were found to be associated with better housing outcomes. As one might expect, homelessness was infrequently resolved during an acute inpatient hospitalization, but few veterans returned to literal homelessness at the time of discharge. PMID- 16702726 TI - Providing health care services to the formerly homeless: a quasi-experimental evaluation. AB - We performed a quasi-experiment to assess the effects of a focused health care intervention delivered in transitional housing facilities in Sacramento, California. Four transitional housing facilities (THFs) receiving the intervention were compared with 2 THFs that did not receive it. A multidisciplinary team provided a diverse package of services to residents at the intervention sites. Residents at comparison sites received usual care. Survey and physical examination data were collected in repeated cross-sectional surveys at baseline (pre-intervention) and after 6 and 18 months of follow-up (post intervention). Using analysis of covariance techniques, our statistical models showed improved odds of receiving recommended gynecologic preventive care and decreased odds of frequent Emergency Department use at 18 months among residents at the intervention sites. At 6 months, residents at the intervention sites also experienced improved blood pressure control. There was no intervention effect on residents' access to specialists or on physical functioning, mental health, or dental health. PMID- 16702728 TI - Drug-metabolizing ability of molybdenum hydroxylases. AB - Molybdenum hydroxylases, which include aldehyde oxidase and xanthine oxidoreductase, are involved in the metabolism of some medicines in humans. They exhibit oxidase activity towards various heterocyclic compounds and aldehydes. The liver cytosol of various mammals also exhibits a significant reductase activity toward nitro, sulfoxide, N-oxide and other moieties, catalyzed by aldehyde oxidase. There is considerable variability of aldehyde oxidase activity in liver cytosol of mammals: humans show the highest activity, rats and mice show low activity, and dogs have no detectable activity. On the other hand, xanthine oxidoreductase activity is present widely among species. Interindividual variation of aldehyde oxidase activity is present in humans. Drug-drug interactions associated with aldehyde oxidase and xanthine oxidoreductase are of potential clinical significance. Drug metabolizing ability of molybdenum hydroxylases and the variation of the activity are described in this review. PMID- 16702729 TI - Construction of several human-derived stable cell lines displaying distinct profiles of CYP3A4 induction. AB - Cell lines which stably express reporter proteins through CYP3A4 gene activation have been developed for use in predicting CYP3A4 induction. Twelve clones showing distinct profiles on chemical-induced response were isolated. Among them, two clones showing high response for CYP3A4 inducers, namely clone 3-1-10 and 3-1-20, were further evaluated for their sensitivities, reproducibilities and applicabilities to predict CYP3A4 induction in human. Clone 3-1-10 showed higher response to rifampicin than to clotrimazole, whereas clone 3-1-20 had rather higher response to clotrimazole. Optimal plating density and highly reproducible response were observed at the range of 1.65-5.0 x 10(4) cell/cm2. Clear induction responses of more than ten chemicals were observed in both cell lines. The reporter activity was further dramatically increased after an introduction of human PXR. Induction with rifampicin was, however, not much altered between the absence and presence of hPXR. The luciferase activity remained unaltered and showed little fluctuation during the culture for more than 6 months. Due to the strikingly high sensitivity and reproducibility of this system, as compared to previously published systems, these HepG2-derived cell lines showing distinct response profiles as developed in the present study will offer high advantages for chemical screening of CYP3A4 inducibility. PMID- 16702730 TI - Genetic variation and haplotype structure of the ABC transporter gene ABCG2 in a Japanese population. AB - The ATP-binding cassette transporter, ABCG2, which is expressed at high levels in the intestine and liver, functions as an efflux transporter for many drugs, including clinically used anticancer agents such as topotecan and the active metabolite of irinotecan (SN-38). In this study, to elucidate the linkage disequilibrium (LD) profiles and haplotype structures of ABCG2, we have comprehensively searched for genetic variations in the putative promoter region, all the exons, and their flanking introns of ABCG2 from 177 Japanese cancer patients treated with irinotecan. Forty-three genetic variations, including 11 novel ones, were found: 5 in the 5'-flanking region, 13 in the coding exons, and 25 in the introns. In addition to 9 previously reported nonsynonymous single nucleotide polymorphisms (SNPs), 2 novel nonsynonymous SNPs, 38C>T (Ser13Leu) and 1060G>A (Gly354Arg), were found with minor allele frequencies of 0.3%. Based on the LD profiles between the SNPs and the estimated past recombination events, the region analyzed was divided into three blocks (Block -1, 1, and 2), each of which spans at least 0.2 kb, 46 kb, and 13 kb and contains 2, 24, and 17 variations, respectively. The two, eight, and five common haplotypes detected in 10 or more patients accounted for most (>90%) of the haplotypes inferred in Block -1, Block 1, and Block 2, respectively. The SNP and haplotype distributions in Japanese were different from those reported previously in Caucasians. This study provides fundamental information for the pharmacogenetic studies investigating the relationship between the genetic variations in ABCG2 and pharmacokinetic/pharmacodynamic parameters. PMID- 16702731 TI - Delayed effect of grapefruit juice on pharmacokinetics and pharmacodynamics of tacrolimus in a living-donor liver transplant recipient. AB - Tacrolimus is a calcineurin inhibitor that has been widely used to prevent allograft rejection after transplantation. We report a case of a living-donor liver transplant recipient experiencing a considerable increase in the trough blood concentration of tacrolimus after concomitant ingestion of grapefruit juice (250 mL) 4 times for 3 days. The trough blood concentrations of tacrolimus were not changed during or immediate after the repeated intake of grapefruit juice. However, almost 1 week after the final ingestion, the blood concentration of tacrolimus markedly increased to as much as 47.4 ng/mL from 4.7 ng/mL before the ingestion, resulting in a profound reduction of calcineurin phosphatase activity in peripheral blood mononuclear cells. Furthermore, headache and nausea, but not nephrotoxicity or hyperglycemia, took place throughout the period of the elevated blood concentrations. Grapefruit juice may have a clinically significant effect on the pharmacokinetics and pharmacodynamics of tacrolimus. It is recommended to avoid the consumption of grapefruit juice in transplant recipients treated with tacrolimus. PMID- 16702732 TI - MDR1 haplotype frequencies in Japanese and Caucasian, and in Japanese patients with colorectal cancer and esophageal cancer. AB - The genotype frequencies of MDR1 T-129C, C1236T, G2677A,T and C3435T SNPs were compared in 154 healthy Japanese and 100 healthy Caucasians to provide basic information on the inter-ethnic differences of pharmacotherapeutic outcome. The variants were found at allelic frequencies of 5.5%, 65.6%, 16.6%, 40.6% and 40.6%, for T-129C, C1236T, G2677A, G2677T and C3435T, respectively, in Japanese, and at 5.1%, 45.9%, 3.6%, 46.4% and 56.6%, respectively, in Caucasians, with a statistically significant difference for C1236T, G2677A,T and C3435T (p<0.001). G2677A was about 5-fold more frequent in Japanese than Caucasians. These genotype frequencies were also investigated in 95 Japanese patients with colorectal cancer (CRC) and esophageal squamous cell carcinoma (ESCC), but no significant difference was detected, when compared with healthy Japanese subjects. The haplotype frequency reached a total of about 85% in Japanese with the following 4 major haplotypes; T(-129)-T1236-T2677-T3435 (36.1%), T(-129)-T1236-G2677-C3435 (22.5%), T(-129)-C1236-G2677-C3435 (14.2%) and T(-129)-C1236-A2677-C3435 (13.3%). The second and fourth haplotypes were hardly inferred in Caucasian, whereas T( 129)-C1236-G2677-T3435 (12.8%) was found to be Caucasian-specific. There was a tendency for higher frequencies of the T(-129)/C-(129)-C1236-A2677-C3435 haplotype in Japanese CRC patients and T(-129)-T1236-T2677-T3435 haplotype in Japanese ESCC patients, compared with that in healthy Japanese subjects. PMID- 16702733 TI - Population pharmacokinetics of pranlukast hydrate dry syrup in children with allergic rhinitis and bronchial asthma. AB - This study aimed to assess the steady-state pharmacokinetics of pranlukast, a leukotriene receptor antagonist, in children with allergic rhinitis and bronchial asthma, and to clarify factors affecting apparent clearance (CL/F). A total of 192 plasma samples were obtained from 98 children (rhinitis 64, asthma 13, complications 21), aged 3-14 years in 2 clinical trials. Plasma concentration of pranlukast was determined by liquid chromatography connected with a tandem mass spectrometer and analyzed by a population approach using NONMEM program. The plasma concentration-time course of pranlukast was described by using a one compartment model with the first-order absorption and lag time. The robustness of the population pharmacokinetic model was evaluated by using 200 bootstrap samples. The results of population pharmacokinetic analysis showed that only age was a factor affecting the CL/F per body weight, with CL/F decreasing with increasing age. No significant variation was seen in the CL/F between rhinitis and asthma. The interindividual variability in the CL/F and the residual variability were 19.7% and 48.4%, respectively. All the parameters fell within 10% of the bootstrapped mean. In conclusion, the results show that age is the most influential factor for explaining interindividual variability in CL/F, and the difference in diseases does not affect CL/F. PMID- 16702734 TI - The structural and pharmacokinetic properties of oxidized human serum albumin, advanced oxidation protein products (AOPP). AB - To determine the pharmacokinetic properties of advanced oxidation protein products (AOPP), we prepared oxidized human serum albumin (oxi-HSA) using chloramine-T (a hypochlorite analogue) in vitro. The AOPP and dityrosine content of oxi-HSA (AOPP content, 244.3+/-12.3 microM; dityrosine content, 0.7+/-0.11 nmol of dityrosine/mg protein) were similar to those of uremic patients. In structural analysis, the increases in AOPP and dityrosine content of HSA induced slight decreases in its alpha-helical content. In pharmacokinetic analysis, oxi HSA left the circulation rapidly, and organ distribution of oxi-HSA 30 min after intravenous injection was 51% for the liver, 23% for the spleen, and 9% for the kidney, suggesting that the liver and spleen were the main routes of plasma clearance of oxi-HSA. The liver and spleen uptake clearance of oxi-HSA were significantly greater than those of normal HSA (CLliver, 5058+/-341.6 vs 24+/-4.2 microL/hr [p<0.01]; CLspleen, 2118+/-322.1 vs 32+/-2.7 microL/hr [p<0.01]). However, uptake by other organs was not significantly affected by oxidation. These results suggest that the liver and spleen play important roles in elimination of AOPP. PMID- 16702735 TI - Involvement of human blood arylesterases and liver microsomal carboxylesterases in nafamostat hydrolysis. AB - Metabolism of nafamostat, a clinically used serine protease inhibitor, was investigated with human blood and liver enzyme sources. All the enzyme sources examined (whole blood, erythrocytes, plasma and liver microsomes) showed nafamostat hydrolytic activity. V(max) and K(m) values for the nafamostat hydrolysis in erythrocytes were 278 nmol/min/mL blood fraction and 628 microM; those in plasma were 160 nmol/min/mL blood fraction and 8890 microM, respectively. Human liver microsomes exhibited a V(max) value of 26.9 nmol/min/mg protein and a K(m) value of 1790 microM. Hydrolytic activity of the erythrocytes and plasma was inhibited by 5, 5'-dithiobis(2-nitrobenzoic acid), an arylesterase inhibitor, in a concentration-dependent manner. In contrast, little or no suppression of these activities was seen with phenylmethylsulfonyl fluoride (PMSF), diisopropyl fluorophosphate (DFP), bis(p-nitrophenyl)phosphate (BNPP), BW284C51 and ethopropazine. The liver microsomal activity was markedly inhibited by PMSF, DFP and BNPP, indicating that carboxylesterase was involved in the nafamostat hydrolysis. Human carboxylesterase 2 expressed in COS-1 cells was capable of hydrolyzing nafamostat at 10 and 100 microM, whereas recombinant carboxylesterase 1 showed significant activity only at a higher substrate concentration (100 microM). The nafamostat hydrolysis in 18 human liver microsomes correlated with aspirin hydrolytic activity specific for carboxylesterase 2 (r=0.815, p<0.01) but not with imidapril hydrolysis catalyzed by carboxylesterase 1 (r=0.156, p=0.54). These results suggest that human arylesterases and carboxylesterase 2 may be predominantly responsible for the metabolism of nafamostat in the blood and liver, respectively. PMID- 16702736 TI - Endogenous uremic substances are not involved in the reduced hepatic extraction of metoprolol in bilateral ureter-ligated rats. AB - The hepatic extraction of metoprolol is reduced in rats with bilateral ureter ligation (BUL)-induced renal failure. The aim of the present study was to evaluate the effect of uremic substances on the hepatic metabolism of metoprolol in rats with BUL. The metabolic rate in the liver microsomes of BUL rats was similar to that in sham rats, and there was no significant difference between sham and BUL rats in the effect of the supernatant of liver homogenates on the metabolism. The rate of metabolism in the liver microsomes in the presence of the plasma of BUL rats was also similar to that in the presence of the plasma of sham rats. These findings indicated that uremic substances which accumulate in BUL rats do not directly inhibit the activity of CYP2D2, which is responsible for the metabolism of metoprolol in the rat liver. PMID- 16702737 TI - Polymorphisms and linkage disequilibrium of the OATP8 (OATP1B3) gene in Japanese subjects. AB - OATP8, a member of the organic anion-transporting polypeptide family, is expressed on the sinusoidal membrane of hepatocytes, and transports endogenous organic anions, such as 17beta-glucuronosyl estradiol, and xenobiotic substances, such as digoxin. The objective of this study is to search for polymorphisms of the OATP8 gene and to assess the allele frequency of the polymorphisms in the Japanese population. Analysis of the OATP8 gene in 79 subjects revealed complete linkage of two deletion polymorphisms in the 5' regulatory region, deletion from position -28 to -11 and deletion from position -7 to -4, with an allele frequency of 0.196 for the deletion allele. The polymorphisms T334G (Ser112Ala) and G699A (Met233Ile) were also shown to be in complete linkage disequilibrium, with an allele frequency of 0.728 for the variant (112Ala/233Ile) allele. Interestingly, linkage disequilibrium was identified between the ins/del polymorphism and SNPs at 112 and 233. The predicted major haplotype was the insert-variant type with a haplotype frequency of 0.60. PMID- 16702738 TI - New modalities for the treatment of acute cardioembolic stroke. PMID- 16702739 TI - ANCA in atheroembolism; just a coincidence or bearing cause and effect? PMID- 16702741 TI - Association between risk factors and carotid enlargement. AB - OBJECTIVE: The aim of this study was to evaluate the influence of conventional cardiovascular risk factors on the degree of adaptive response of the carotid arterial wall to atherosclerotic disease. PATIENTS AND METHODS: We evaluated the diameter and intima-media thickness (IMT) of common carotid artery (CCA) by ultrasonography in 351 men aged 70.3 (range, 14-97) years and 474 women aged 75.6 (range, 19-103) years in the medical department of Seiyo Municipal Nomura Hospital. We assessed cross-sectionally the relationships between CCA diameter and IMT and cardiovascular risk factors by gender. RESULTS: In multiple linear regression analyses, after controlling for traditional cardiovascular risk factors, a significant correlation was found between CCA diameters and age (p=0.034), body mass index (BMI) (p<0.001), smoking status (p=0.039), alcohol consumption (<0.001) and uric acid (UA) (p=0.021) in men, and between CCA diameters and age (<0.001), BMI (p<0.001), systolic blood pressure (SBP) (p=0.013) and antihypertensive drug use (p=0.005) in women. Analysis of covariance showed that the two regression lines between carotid IMT and diameter in those with or without plaque were significantly different in both men (F=16.4; p<0.001) and women (F=15.0; p<0.001). After adjustment for carotid IMT and plaque, associations with carotid diameters still persisted for age (p<0.001), BMI (p<0.001), smoking status (p=0.006), alcohol consumption (p<0.001) and SBP (p=0.001) in men, and age (p=0.005), BMI (p<0.001), SBP (p=0.047) and UA (p=0.001) in women. CONCLUSION: This study shows that the CCA diameters correlated with conventional cardiovascular risk factors including alcohol consumption. These findings suggest that the CCA diameters may reflect the ability of adaptive remodeling to the atherosclerosis before plaque formation and can be an important factor during the development of atherosclerosis. PMID- 16702740 TI - Long-term outcome of autoimmune pancreatitis after oral prednisolone therapy. AB - OBJECTIVE: We investigated the long-term outcome of autoimmune pancreatitis (AIP) including morphological changes in the pancreas, pancreatic duct, biliary tract, pancreatic function, and changes in the clinical manifestations after oral prednisolone (PSL) therapy. PATIENTS AND METHODS: We prospectively followed 12 patients for a period of over 12 months (median follow-up period: 41 months; range: from 13 to 133 months). All twelve patients were treated with PSL. The morphological findings consisted of pancreatic enlargement (n=12), an irregularly narrowed main pancreatic duct (n=12), and bile duct stricture (n=10), and salivary gland swelling was observed in six patients. The initial dose of PSL was 30-40 mg/day, and it was subsequently tapered. RESULTS: All 12 patients responded to PSL therapy. The enlargement of the pancreas and the irregularly narrowed main pancreatic duct improved to almost normal. Pancreatic atrophy developed in four of them (4/12, 33%), but no pancreatic calcification was observed in any of the patients. The bile duct stricture improved to various degrees in all 10 patients , but it persisted in the lower part of the bile duct in four of them (4/10, 40%). The salivary gland swelling also improved after PSL therapy. There was no recurrence of enlargement of the pancreas or irregularly narrowed main pancreatic duct after PSL therapy, but the bile duct stricture recurred in one case, and in three cases there was a relapse of salivary gland swelling that required a temporary increase in PSL dose during tapering. No deterioration of pancreatic exocrine function was detected in any of the patients. A malignant tumor was diagnosed in two patients during PSL therapy: early gastric cancer in one and rectal cancer in the other. All patients are alive. CONCLUSIONS: AIP treated with PSL has a favorable long-term outcome based on the morphological findings and assessments of pancreatic function. However, since two of the twelve patients developed a malignancy during PSL therapy, strict follow up should be part of the management of AIP. PMID- 16702742 TI - Recruitment of patients for a clinical trial: factors on the physician side and reasons on the patient side. AB - OBJECTIVES: To examine the factors related to actual patient recruiters among the physicians who initially agreed to collaborate in a randomized control trial. METHODS: We conducted a questionnaire survey of 679 physicians (512 actual recruiter and 167 non-recruiters) who had initially agreed to recruit patients for a clinical trial to determine factors to predict who would actually do so. RESULTS: Response rates among recruiters and non-recruiters were 87.5% and 73.1%, respectively. Multivariate logistic regression model showed that the proportions of regular users of computer [odds ratio (OR) =2.1, 95% confidence intervals (CI)=1.3-3.3] (p=0.002) and current participants in other clinical trials (OR=2.2, CI=1.5-3.4) (p=0.001) were significantly higher among recruiters than non-recruiters. Patients' reasons for non-participation as perceived by the physicians did not differ between recruiters and non-recruiters. CONCLUSION: Results of this study might be useful in predicting actual recruiters at the outset of clinical trials. PMID- 16702743 TI - A Japanese case of familial Mediterranean fever with onset in the fifties. AB - The patient was a 63-year-old woman with attacks of fever and abdominal pain, starting from the age of 53 years and recurring every month. Despite various examinations at another hospital, the etiology remained unclear. She was under symptomatic treatment, and was referred to our department for further evaluation. Although she had onset in middle age, the clinical symptoms and examination findings suggested familial Mediterranean fever, and administration of colchitine inhibited the attacks completely. Therefore, the patient was diagnosed as having the disease. We were not able to analyze the entire MEFV gene, but detected only a heterozygous M694I mutation. Amyloidosis did not develop as a complication. The disease is rare in Japan, and its onset in the fifties is extremely rare in the world. PMID- 16702744 TI - Glucose-responsive and octreotide-sensitive insulinoma. AB - Insulinoma is the most common cause of fasting hypoglycemia resulting from autonomous insulin hypersecretion. We describe herein a unique case with paradoxical hypoglycemic episodes induced by hyperglycemia. A 55-year-old female had repeated hypoglycemic episodes after meal or during increased physical activity. Although fasting (10 hr) failed to provoke hypoglycemia, oral glucose tolerance test (GTT) caused an exaggerated insulin response (885 microU/ml) at 30 min, followed by hypoglycemia (36 mg/dl) after 90 min. Moreover, intravenous GTT also induced an exaggerated insulin response (>2900 microU/ml) at 10 min, followed by hypoglycemia (34 mg/dl) after 40 min. Although MRI and CT scan of the abdomen failed to detect any mass lesions in the pancreas, Octreoscan revealed increased radioactive uptake around the pancreatic head region. Treatment with a daily injection of octreotide (100 microg) alleviated her hypoglycemic episodes. At surgery, two islet cell adenomas were identified in the pancreas and resected. Postoperatively, she was free from hypoglycemic episodes after meal. Postoperative oral and intravenous GTT did not induce hypoglycemia. Thus, this is a very rare case of glucose-responsive and octreotide-sensitive insulinoma in whom GTT and octreotide proved to be a useful provocation and treatment for hypoglycemic episodes. PMID- 16702745 TI - Glucocorticoid therapy ameliorated hypoglycemia in insulin-like growth factor-II producing solitary fibrous tumor. AB - A 54-year-old man with a past history of multi-operation for solitary fibrous tumor was hospitalized for hypoglycemia. Computed tomography demonstrated multiple tumors in the liver but not in the pancreas. His serum insulin-like growth factor (IGF)-II level was normal, but immunoblot analysis and immunohistochemistry revealed a high molecular weight form of IGF-II in the serum and tumor. Surgical resection was impossible, because of tumor dissemination. Hypoglycemia was repeated despite infusion of glucose and glucagon. Glucocorticoid dramatically and continuously abolished hypoglycemia. In cases of inoperable IGF-II-producing tumor, glucocorticoid therapy may be promising for hypoglycemia. PMID- 16702746 TI - A case with membranous lupus nephritis developing after a twenty-year remission of membranoproliferative glomerulonephritis. AB - A 30-year-old woman who showed remission of membranoproliferative glomerulonephritis (MPGN) 20 years previously developed membranous lupus nephritis (MLN). She had photosensitivity, facial erythema, proteinuria of 2.59 g/24 hr, anti-nuclear antibody and anti-ds-DNA antibody. To confirm whether a misdiagnosis of MPGN was made 20 years ago, the clinical data at that time were evaluated retrospectively. She had only mild proteinuria and hematuria but no photosensitivity or facial erythema. Anti-nuclear antibody was negative. Renal biopsy showed occasional lobulation and glomerular capillary double contour. The diagnosis of MPGN was definite. This might be a rare case of one person suffering from two types of glomerulonephritis, MPGN and MLN. PMID- 16702747 TI - Hypersensitivity pneumonitis induced by spores of Penicillium citrinum in a worker cultivating Enoki mushroom. AB - A 47-year-old Japanese woman was admitted to our hospital with a 2-week history of dry cough and shortness of breath. She had been engaged in Enoki mushroom production for 22 years. Chest X-ray and chest computed tomography (CT) scan showed bilateral fine-nodular shadows and ground glass opacity. Bronchoalveolar lavage fluid demonstrated an increase of total cell counts with predominant lymphocytosis. Pathological specimens obtained by video-assisted thoracoscopic surgery revealed alveolitis and noncaseating granuloma with giant cells. Lymphocyte stimulation test showed positive responses with Enoki mushroom, culture medium, and Penicillium citrinum. On double immunodiffusion test, a precipitation line was observed between patient's serum and Penicillium citrinum antigen. She was found to have hypersensitivity pneumonitis caused by Penicillium citrinum. This is the first report of mushroom worker's lung caused by Penicillium citrinum. PMID- 16702748 TI - Pulmonary adenocarcinoma associated with SAPHO syndrome difficult to differentiate from multiple bone metastasis. AB - The patient was a 57-year-old man with a chief complaint of anterior chest pain who was diagnosed with clinical stage IV (c-T2N2M1) non-small-cell lung cancer (adenocarcinoma). Tenderness in the sternoclavicular joint, acne, periodontitis, and palmoplantar pustulosis were evident, and SAPHO syndrome was diagnosed. SAPHO syndrome is a rare disorder that results in synovitis, acne, pustulosis, hyperostosis, and osteomyelitis. Bone scintigraphy showed tracer accumulation in the costal cartilage, sternoclavicular joint, and cervical vertebrae 6-7. Although the bone lesions of SAPHO syndrome were difficult to differentiate from bone metastasis of pulmonary adenocarcinoma, metastatic bone tumors were ruled out by magnetic resonance imaging, computed tomography, and fluorodeoxyglucose positron emission tomography. There have been no previously reported cases of lung cancer with comorbid SAPHO syndrome. We report such a case and discuss the relevant literature, particularly that concerned with the evaluation of bone lesions. PMID- 16702749 TI - Myocardial infarction caused by Aspergillus embolization in a patient with aplastic anemia. AB - A 38-year-old Japanese man with severe aplastic anemia had invasive pulmonary aspergillosis as a complication. He was treated with amphotericin B for six weeks, but the aspergillosis did not improve. Then he experienced a fatal myocardial infarction. An autopsy revealed disseminated aspergillosis involving pericarditis and Aspergillus embolization to the coronary arteries. This led to the acute myocardial infarction. Cardiac aspergillosis is rare, but should be included within the differential diagnosis when chest pain of unknown origin occurs in an immunosuppressed patient. PMID- 16702750 TI - Aseptic meningoencephalitis presenting with bilateral vestibular ataxia: a case report. AB - Bilateral vestibular dysfunction is a rare condition, of which peripheral disorders are most common, whereas central disorders are extremely rare. A 35 year-old woman developed fever, headache, dizziness, convulsion, and disturbance of consciousness at the same time. MRI findings were normal. Based on the neurological findings and laboratory abnormalities, she was diagnosed as having aseptic meningoencephalitis. Steroid pulse therapy successfully ameliorated her encephalitis sign. However, isolated bilateral vestibular ataxia and dizziness, together with severe decrease of Caloric nystagmus, became apparent and lasted for the following 3 months, without cerebellar/brainstem involvement. PMID- 16702751 TI - Convergence substitution in pontine tegmental hemorrhage: an observation with Frenzel goggles. AB - We report a case in which convergence movement was substituted for bilateral horizontal gaze palsy due to a pontine tegmental hemorrhage. When visual fixation was suppressed with Frenzel goggles (FG), substituted convergence movements were prominent in our patient. The observation suggests that both vergence eye movements and horizontal gaze movements act synergistically and lesioning of the version system results in overactivity of the convergence system after a pontine hemorrhage. Furthermore, it may be worth using FG in the observation of substituted eye movements in patients with horizontal gaze palsy due to a pontine tegmental injury. PMID- 16702753 TI - Insidious phrenic nerve involvement in postpolio syndrome. AB - A 49-year-old woman with amyotrophic sequelae of poliomyelitis experienced progressive left upper limb weakness and breathing discomfort while walking that had developed over one year prior to presentation. She had flaccid quadriplegia which was more marked in the left upper limb, with the C4- and C5-innervated muscles being most severely affected. Chest radiographs double exposed at maximal inspiration and expiration revealed poor respiratory movements in the left hemidiaphragm. Phrenic nerve conduction study demonstrated conspicuous diaphragmatic paralysis on the left side. The electrophysiological study supports radiological findings and provides useful evidence for insidious phrenic nerve involvement in postpolio syndrome. PMID- 16702752 TI - Systemic cholesterol embolization syndrome associated with myeloperoxidase-anti neutrophil cytoplasmic antibody. AB - A 75-year-old man was transferred to our department because of development of severe renal impairment after coronary artery bypass grafting. Hemodialysis was initiated for postsurgical oliguria and lung congestion. On transfer, he showed systemic purpura rashes and diffuse blue mottlings on his toes with marked eosinophilia and an elevated level of C-reactive protein. Cutaneous biopsy revealed cholesterol crystal embolism and leukocytoclastic vasculitis in dermal arterioles. Myeloperoxidase-anti-neutrophil cytoplasmic antibody titer was increased. Upon oral corticosteroid therapy following intravenous pulse steroid therapy, the purpura dramatically diminished, renal function improved, and hemodialysis was discontinued. Active treatment with corticosteroids may be effective for cholesterol embolization syndrome, particularly when clinical and laboratory manifestations mimic systemic vasculitis. PMID- 16702755 TI - Bilateral diffuse enlargement of the kidneys after exercise. PMID- 16702754 TI - Hydatid cysts in the liver. PMID- 16702756 TI - Nasal T-cell lymphoma. PMID- 16702757 TI - Calcium ion as a second messenger with special reference to excitation contraction coupling. AB - Calcium ion (Ca(2+)) plays an important role in stimulus-response reactions of cells as a second messenger. This is done by keeping cytoplasmic Ca(2+) concentration low at rest and by mobilizing Ca(2+) in response to stimulus, which in turn activates the cellular reaction. The role of Ca(2+) as a second messenger was first discovered in excitation-contraction coupling of skeletal muscle. The history of the discovery was reviewed. Characteristics of Ca(2+) as a second messenger, diversity of target molecules, capability of rapid and massive mobilization and also of oscillatory mobilization, tendency toward localization, and on the other side, ability to cause generalized cell response were described. The possible bases for these characteristics was discussed. Ca(2+) itself induces release of Ca(2+) from the sarcoplasmic reticulum (Ca(2+)-induced Ca(2+) release [CICR]). The Ca(2+) release channel, ryanodine receptor, incorporated into lipid bilayer shows CICR activity. Ca(2+) release induced by inositol trisphosphate also has an apparent CICR nature. The significance of CICR or apparent CICR with its inherently regenerative nature in physiological contractions of skeletal, cardiac, and smooth muscles was discussed. PMID- 16702758 TI - Seasonal changes in immunolocalization of inhibin/activin subunits and testicular activity in wild male raccoon dogs (Nyctereutes procyonoides). AB - Thirty-four pairs of testes from wild adult raccoon dogs (Nyctereutes procyonoides) were obtained between September 2000 and May 2003. The cellular localization of the inhibin alpha and inhibin/activin (betaA and betaB) subunits in wild raccoon dog testes was investigated. The testicular weight and size and seminiferous tubule diameters were measured. There were marked seasonal variations in testicular weight and size and seminiferous tubule diameters, with values relatively low in September and high in March. Spermatogonia and primary spermatocytes were observed in September, and spermatogonia, spermatocytes, and round spermatids were present in January. All types of spermatogenic cells, including mature spermatozoa, were found in March, indicating that the breeding season is around March in Japan. Thereafter, spermatogonia and degenerating spermatocytes were observed in April. The sections of testes were immunostained by the avidin-biotin-peroxidase complex method (ABC) using polyclonal antisera raised against porcine inhibin alpha, inhibin/activin betaA and inhibin/activin betaB. The inhibin alpha and inhibin/activin (betaA and betaB) subunits were only expressed in Leydig cells in September. On the other hand, the inhibin alpha, betaA, and betaB subunits were observed in Leydig cells and Sertoli cells, but not in germ cells, in March. These results suggest that the testes of wild raccoon dogs have the ability to synthesize inhibins, and the cellular localization of inhibin/activin subunits showed season-related changes in the breeding and non-breeding seasons. PMID- 16702759 TI - The relationship of TRP channels to the pacemaker activity of interstitial cells of Cajal in the gastrointestinal tract. AB - Interstitial cells of Cajal (ICCs) are a fundamental component of the pacemaker apparatus of the gastrointestinal (GI) tract. They have special properties that make them unique in their ability to generate and propagate slow waves in gastrointestinal smooth muscle. The pacemaker current that generates slow waves is initially due to a voltage-independent, Ca(2+)-inhibited, non-selective cationic conductance in ICC. The classical transient receptor potential (TRPC) channel 4 was suggested as a molecular candidate for the nonselective cation channel (NSCC) responsible for the pacemaker activity. We have shown that TRPC4-/ mice display normal slow waves and suggest that TRPC4 might be an essential component of the NSCC activated by muscarinic stimulation. Finally, we suggest that TRPM7 is the molecular candidate for the NSCC responsible for pacemaker activity in ICCs on the basis of electrophysiological, molecular biological, and immunohistochemical experiments. PMID- 16702760 TI - Correlations between the responses of electrogastrograms, heart rate and respiratory rate to the stress of the mirror drawing test in human subjects. AB - An electrogastrogram (EGG) recorded on the skin is believed to reflect the electrical activity of the gastrointestinal tract. In this study, linear correlations were calculated for the EGG spectral frequencies and the power ratios, changes in heart and respiratory rate before and during mirror drawing test (MDT) stress and after having meals, with the changes in the sympathetic tone indicator (LF/HF) and HADS anxiety and depression scores in human subjects (n=48). Depression scores had a negative linear correlation with the EGG resting frequencies of the epigastric and supraumbilical 6 cpm groups (5.0-7.4 cpm). Anxiety scores had a positive linear correlation with the EGG area power ratio of the resting to MDT stress responses of the epigastric and infraumbilical 6 cpm groups (6 cpm area during MDT/that during fasting rest). The resting LF/HF had a positive linear correlation with the EGG area power ratio of the resting to MDT stress responses of the epigastric and infraumbilical 3 cpm groups (2.5-4.9 cpm). The resting LF/HF ratio had a negative correlation with the area power ratio of the resting to postprandial responses (postprandial power/resting power) in the epigatric, supraumbilical and infraumbilical 6 cpm groups. The resting LF/HF ratio had a positive linear correlation with the heart rate at rest and during MDT-stress. In contrast, the resting LF/HF ratio had a negative linear correlation with the respiratory rate at rest and during MDT-stress. EGGs of the surface electrical activities of the gastrointestinal tract were influenced by acute MDT-stress and their frequencies and their response power ratios were correlated linearly with anxiety scores, depression scores and the sympathetic tone indicator, the LF/HF ratio. PMID- 16702761 TI - B(2)-receptor modulation of the reactivity to phenylephrine and angiotensin II in the carotid artery of normotensive rats after trandolapril treatment. AB - This study was designed to study the effects of angiotensin converting enzyme inhibitors (ACEI) following treatment with trandolapril (0.3 mg kg(-1) day(-1)) on carotid arterial responsiveness in normotensive Wistar rats. Carotid arteries were obtained from control or trandolapril-treated animals and mounted in an isolated organ bath. Reactivity to angiotensin II (Ang II), phenylephrine (Phe) and KCl was studied. Agonist concentration-response curves were constructed in either the absence or presence of the endothelium or after incubation with L-NAME (10(-6) M), HOE140 (10(-7) M) or indomethacin (10(-5) M). Trandolapril treatment decreased the Ang II and Phe potencies in carotid arteries, but did not affect the maximal response. The KCl responses (potency and Emax) were similar in both control and trandolapril-treated arteries. The absence of endothelium increased the response to both agonists in control and trandolapril-treated arteries; however, the inhibitory component from the endothelial layer of the Phe response was greater in trandolapril-treated animals than in control animals. The presence of L-NAME or HOE140 abolished the changes in the potency values of trandolapril treated animals. The presence of indomethacin did not change the effect of trandolapril on the potency values of both agonists. We conclude that trandolapril treatment decreased the carotid arterial reactivity in normotensive rats and that this effect is endothelium-dependent. Furthermore, the involvement of B(2)-receptors and NO production, but not of prostaglandins, is suggested in this mechanism. PMID- 16702762 TI - Metabolic component of the temperature-sensitivity of slow waves recorded from gastric muscle of the guinea-pig. AB - The effects of changes in temperature on slow waves were investigated in smooth muscle tissues isolated from the guinea-pig gastric antrum. Within the range 24 degrees C to 42 degrees C, elevation of temperature increased the frequency and maximum rate of rise of the upstroke phase (dV/dt) of slow waves and decreased their duration, with no alteration to amplitude or resting membrane potential. These observations also applied to follower potentials and pacemaker potentials recorded from longitudinal muscle and myenteric interstitial cells, respectively. Slow waves were comprised of 1st and 2nd components, and the latency for generating the 2nd component was decreased exponentially by elevating temperature, reaching a stable value of about 1 s above 32 degrees C. The temperature coefficient was >2 for the frequency, dV/dt and latency of the 2nd component, about 1.7 for the duration and about 1 for amplitude. Potassium cyanide (KCN), an inhibitor of mitochondrial metabolic activity, reduced the frequency and duration of slow waves, with no alteration to other parameters (amplitude, dV/dt, latency). In the presence of 30 microM KCN, the temperature dependency of the frequency of slow waves was diminished or abolished, while other parameters of slow waves remained unaltered. These results indicate that in slow waves the frequency may be related to metabolic activities, while the temperature-dependent changes in the dV/dt, latency for the 2nd component and duration of slow waves are produced largely by mechanisms other than metabolic activity. PMID- 16702763 TI - Effect of heptanol on noradrenaline-induced contractions in rat vas deferens. AB - We have studied the effects of 1-heptanol and nifedipine on noradrenaline (NA) induced contractions in order to explore the role of gap junctions and their interactions with L-type Ca2+ channel mediated [Ca2+]o entry in the generation of NA-induced contractions in the rat vas deferens. Application of 20 microM NA to rat vas deferens resulted in contractions with three different components, an initial phasic component followed by a tonic component overlapped with an oscillatory component. Heptanol (0.01-2 mM) induced a concentration dependent reduction of the contractions. 2 mM heptanol reduced the phasic component by 32.9 +/- 4.4% and the tonic component by 93.8 +/- 1.9% of control, while the oscillatory component was completely abolished (n=7). Nifedipine (2 microM) reduced the phasic component by 34.5 +/- 4.1% and the tonic component by 89.5 +/- 3.8% of control and abolished the oscillatory component (n=6). In the presence of heptanol and nifedipine together, the phasic component was reduced by 61.3 +/- 8.3% and the tonic component by 94.5 +/- 1.0% of control. The oscillatory component was completely abolished (n=6). These results allow the conclusion that phasic contraction is mainly due to the direct action of NA, independent of gap junctions, while the tonic and oscillatory contractions may depend significantly on cell-to-cell communication. These in turn may depend critically on the availability of extracellularly derived Ca2+. PMID- 16702764 TI - Comparative phylogeography of two bitterlings, Tanakia lanceolata and T. limbata (Teleostei, Cyprinidae), in Kyushu and adjacent districts of western Japan, based on mitochondrial DNA analysis. AB - To understand the geographical patterns of genetic variation in freshwater fishes in western Japan, the genetic structures of populations of Tanakia lanceolata and T. limbata (Teleostei, Cyprinidae) in this area were investigated using mitochondrial DNA (mtDNA) cytb sequences. Neighbor-joining trees of mtDNA haplotypes revealed four and three genetically divergent groups in T. lanceolata and T. limbata, respectively. Each group was restricted to one or the other of the geographical regions in the area studied. The patterns of geographical divergence in the two species showed some similarities, which seem to reflect common historical events experienced by freshwater fishes distributed in western Japan. On the other hand, dissimilarities were also found in the patterns, indicating that species-specific historical processes also occurred. Within one region, T. lanceolata was less differentiated than T. limbata, suggesting a difference in the dispersal abilities of the two bitterlings. In addition, several individuals in Kunichika River on Shikoku island were morphologically identified as T. lanceolata but had mtDNA haplotypes of T. limbata. We tentatively suggest that these individuals are hybrids of the two species, but further studies employing nuclear markers are necessary to validate this hypothesis. PMID- 16702765 TI - Phylogenetic and taxonomic relationships of northern Far Eastern phoxinin minnows, Phoxinus and Rhynchocypris (Pisces, Cyprinidae), as inferred from allozyme and mitochondrial 16S rRNA sequence analyses. AB - Analyses of allozyme (18 loci) and partial mitochondrial DNA (mtDNA) sequences (1295 bp, 16S rRNA) support the classification of phoxinin minnows from the northern Far East into 2 genera of 8 species: Phoxinus phoxinus, Rhynchocypris oxycephalus, R. perenurus, R. czekanowskii, R. kumgangensis, R. semotilus, R. lagowskii and R. sp. (bergi ?). Although R. lagowskii from Japan and the Amur basin and R. sp. from Vladivostok region to Korea have been classified into a single species by many authors as R. lagowskii, they form separate clusters in both analyses, suggesting different specific status. Some R. oxycephalus and R. perenurus had the mtDNA haplotypes of R. lagowskii and R. czekanowskii, respectively, which probably indicates that local introgression of mtDNA occurred through inter-specific hybridization. Rhynchocypris forms a monophyletic cluster with dace genera Tribolodon and Pseudaspius, not with Phoxinus. Eurasian and American Phoxinus are suggested to be paraphyletic. PMID- 16702766 TI - Population structure and gene flow among wild populations of the black muntjac (Muntiacus crinifrons) based on mitochondrial DNA control region sequences. AB - The black muntjac (Muntiacus crinifrons) is a rare species endemic to China and its current distribution is confined to partial mountain ranges in eastern China. To assess the population structure and gene flow among the extant populations, we sequenced a portion of the mitochondrial control region (424 bp) for 47 samples collected from its current three large populations (Huangshan, Tiammushan and Suichang). A total of 18 unique haplotypes were defined based on 22 polymorphic sites. Of these haplotypes, 15 ones were population-specific and only one haplotype was shared among the three populations. Significant genetic differentiation was detected between Suichang and Huangshan populations (o(ST)=0.1677, P<0.001) or between Suichang and Tianmushan populations (o(ST)=0.2002, P<0.001), indicating that the Suichang population may be spatially structured from other two populations along matriline. The Mantel test revealed that this significant differentiation was not driven by geographic distance (P=0.14), implying that genetic divergence of Suichang population might result from recent human disturbances. Phylogenetic analyses suggested the mitochondrial control region haplotypes were split into two well divergent clades (Clade I and Clade II). Interestingly, the two distinct haplotype clades were found to coexist in Suichang area. The nested clade analysis revealed a significant phylogeographic structure among the black muntjac populations (total cladogram: chi2=18.68; P<0.001), which was inferred to result from past fragmentation followed by range expansion. The population expansion was supported by the analysis of mismatch distribution and the tests of neutrality. Therefore, we suggest that the coexistence of distinct haplotypes in Suichang population was induced by historical population expansion after fragmentation and that the current genetic differentiation should be attributed to the reduction of female mediated gene flow due to recent habitat fragmentation and subsequent loss. PMID- 16702768 TI - Behavioral responses to the alarm pheromone of the ant Camponotus obscuripes (Hymenoptera: Formicidae). AB - The alarm pheromone of the ant Camponotus obscuripes (Formicinae) was identified and quantified by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Comparisons between alarm pheromone components and extracts from the major exocrine gland of this ant species revealed that the sources of its alarm pheromone are Dufour's gland and the poison gland. Most components of Dufour's gland were saturated hydrocarbons. n-Undecane comprised more than 90% of all components and in a single Dufour's gland amounted to 19 microg. n-Decane and n-pentadecane were also included in the Dufour's gland secretion. Only formic acid was detected in the poison gland, in amounts ranging from 0.049 to 0.91 microl. This ant species releases a mixture of these substances, each of which has a different volatility and function. When the ants sensed formic acid, they eluded the source of the odor; however, they aggressively approached odors of n undecane and n-decane, which are highly volatile. In contrast, n-pentadecane, which has the lowest volatility among the identified compounds, was shown to calm the ants. The volatilities of the alarm pheromone components were closely related to their roles in alarm communication. Highly volatile components vaporized rapidly and spread widely, and induced drastic reactions among the ants. As these components became diluted, the less volatile components calmed the excited ants. How the worker ants utilize this alarm communication system for efficient deployment of their nestmates in colony defense is also discussed herein. PMID- 16702767 TI - Phylogeny of selected Sepiidae (Mollusca, Cephalopoda) based on 12S, 16S, and COI sequences, with comments on the taxonomic reliability of several morphological characters. AB - Phylogenetic relationships among 11 species of sepiids from Japanese waters and Sepia officinalis from Mediterranean were studied using partial sequences of the mitochondrial 12S rRNA, 16S rRNA, and cytochrome c oxidase subunit I genes. These three genes had been analyzed in an Atlantic species S. elagans and was obtained from database. In the two-gene set analysis (16S+COI), sequence data of another 4 species were added from database. We also studied morphological characters of radulae, tentacular clubs, and cuttlebones. The molecular phylogeny was not congruent with relationships detected by the number of rows in radulae and the arrangement of suckers on the tentacular club. As to the cuttlebone shape, the molecular phylogeny suggests the separation of two groups, Doratosepion species with a lanceolate cuttlebone and the others with a broad cuttlebone. Our molecular phylogenetic study revealed these sepiids are separated into four clades. The first clade includes Sepia officinalis, S. hierrendda, S. bertheloti, S. pharaonis and Sepiella japonica. The second clade consists of S. latimanus and Metasepia tullbergi from sub-tropical waters. The third clade includes Sepia esculenta, S. madokai, S. aculeata and S. lycidas, which have a cuttlebone with a prominent spine. The fourth clade consists of Doratosepion species complex, S. kobiensis, S. lorigera, S. pardex, S. peterseni, and S. sp., which are characterized by a narrow cuttlebone with a distinct outer cone at the posterior end. The lack of membranous structures in the cuttlebone is a synapomorphy for this clade. S. elegans did not clearly belong to any of these clades and might represent the fifth clade. PMID- 16702769 TI - Behavioral analyses of wind-evoked escape of the cricket, Gryllodes sigillatus. AB - The wind-evoked escape behavior of the cricket Gryllodes sigillatus was investigated using an air puff stimulus. A high velocity air puff elicited the escape behavior in many crickets. The crickets tended to escape away from the stimulus source, but the direction was not accurately oriented 180 degrees from the stimulus. After bilateral cercal ablation, only a few crickets showed wind evoked escape behavior, and their response rates did not increase even 19 days after ablation. Therefore, information on air motion detected by cercal filiform hairs is essential for triggering wind-evoked behavior. After unilateral cercal ablation, the 81.3% response rate of intact crickets decreased to 16.5%, that is, it decreased to almost 20% that of intact crickets. One week after unilateral cercal ablation, the response rate recovered to more than 60% that of intact crickets. However, the accuracy rate of the escape direction of G. sigillatus showed no change even immediately after the unilateral cercal ablation. Therefore, both cerci are not necessarily required to determine the escape direction. The behavioral characteristics of wind-evoked escape of G. sigillatus are compared with those of another species of cricket, Gryllus bimaculatus. The two species of cricket employ different strategies for wind-evoked escape. PMID- 16702770 TI - Cercal sensory system and giant interneurons in Gryllodes sigillatus. AB - The external morphologies of two cricket species, Gryllodes sigillatus and Gryllus bimaculatus, were investigated. Despite its small body length, G. sigillatus possessed longer cerci and longer cercal filiform hairs than G. bimaculatus. The estimated number of filiform hairs on a cercus was also larger in G. sigillatus than in G. bimaculatus. Wind-sensitive interneurons receiving sensory inputs from cercal filiform hairs and running in the ventral nerve cord (VNC) were investigated in G. sigillatus both morphologically and physiologically. By intracellular staining, these interneurons were proved to be morphologically homologous with previously identified giant interneurons (GIs 8 1, 9-1, 9-2, 9-3, 10-2, and 10-3) in G. bimaculatus and Acheta domesticus. In G. sigillatus, the intensity-response relationship (I-R curve) for each GI was investigated using a unidirectional air current stimulus. The stimulus was applied from 12 different directions, and an I-R curve was obtained for each stimulus direction. Each GI showed a characteristic I-R curve depending on stimulus direction. The directionality curve expressed in terms of threshold velocity showed that each GI had a distinctive directional characteristic. The functional properties of GIs in G. sigillatus, such as I-R curve, threshold velocity, and directional characteristics, were compared with those of homologous GIs in G. bimaculatus in Discussion. PMID- 16702771 TI - Renewal of genetic composition of a lancelet, Branchiostoma belcheri, in the Seto Inland Sea, Japan. AB - We studied renewal of genetic composition of a lancelet, Branchiostoma belcheri, at a station in the Seto Inland Sea, Japan, using random amplified polymorphic DNA (RAPD) technique. We detected a total of 49 DNA bands from 27 individuals of 5 seasonal samples. Among these, some were specific to individual or seasonal sample. There were no bands common to all individuals. We calculated the Band Sharing Index (BSI) values for each combination of individuals. BSI was in a wide range of 0.00-0.97 and the average BSI between the individuals of a sample was remarkably different from sample to sample (0.37-0.83). In cluster analysis on the basis of BSI, largely 3 clusters were constructed, which finally connected to a single cluster at as low as about 0.2 of BSI. Besides, many clusters were constructed exclusively by a seasonal sample. These results suggest a large gene pool of lancelets in this area, the remarkable differences in gene pool size among the samples and a seasonal renewal of genetic composition at a habitat. PMID- 16702772 TI - Preparation of two recombinant crustacean hyperglycemic hormones from the giant freshwater prawn, Macrobrachium rosenbergii, and their hyperglycemic activities. AB - Crustacean hyperglycemic hormone (CHH) is released from the X-organ/sinus gland complex located in the eyestalks, and regulates glucose levels in the hemolymph. In the giant freshwater prawn (Macrobrachium rosenbergii), two cDNAs encoding different CHH molecules were previously cloned by other workers. One of these (Mar-CHH-2) was expressed only in the eyestalks, whereas the other (Mar-CHH-L) was expressed in the heart, gills, antennal gland, and thoracic ganglion, but not in the eyestalks. However, their biological activities had not yet been characterized. Therefore, in this study, recombinant Mar-CHH-2 (rMar-CHH-2) and Mar-CHH-L (rMar-CHH-L) were produced using an E. coli expression system, by expression in bacterial cells and recovery in the insoluble fraction. Thereafter, rMar-CHH-2 and rMar-CHH-L were subjected to refolding and were subsequently purified by reversed-phase HPLC. The rMar-CHH-2 and rMar-CHH-L thus obtained exhibited the same disulfide bond arrangements as those of other CHHs reported previously, indicative of natural conformation. In in vivo bioassay, rMar-CHH-2 showed significant hyperglycemic activity, whereas rMar-CHH-L had no effect. These results indicate that Mar-CHH-L does not function as a CHH, but may have some other, unknown function. PMID- 16702774 TI - Regulation of brain development by thyroid hormone and its modulation by environmental chemicals. PMID- 16702773 TI - Molecular phylogeny of Cryptocercus wood-roaches based on mitochondrial COII and 16S sequences, and chromosome numbers in Palearctic representatives. AB - Woodroaches of the genus Cryptocercus are subsocial and xylophagous cockroaches, distributed in North America and Asia. Studies on male chromosome number in Nearctic species have shown that diploid numbers vary from 2n=37 to 2n=47; numbers from Palearctic species were heretofore unknown. Two hypotheses have been proposed to explain the varying number of chromosomes among Nearctic species: the serial reduction hypothesis, and the parallel scenario. We performed phylogenetic analyses of the COII gene in these species and found evidence for the topology (47(45(43(39,37), which is congruent with the serial reduction hypothesis. We also determined chromosome numbers for the first time in Palearctic species, and found Cryptocercus primarius and Cryptocercus relictus to have relatively low chromosome numbers (2n=17-21) compared to their Nearctic relatives. Finally, our study determined the phylogenetic position of Cryptocercus primarius among other Asian taxa. PMID- 16702775 TI - Insulin receptor signals regulating GLUT4 translocation and actin dynamics. AB - In skeletal muscle and adipose tissue, insulin-stimulated glucose uptake is dependent upon translocation of the insulin-responsive glucose transporter GLUT4 from intracellular storage compartments to the plasma membrane. This insulin induced redistribution of GLUT4 protein is achieved through a series of highly organized membrane trafficking events, orchestrated by insulin receptor signals. Recently, several key molecules linking insulin receptor signals and membrane trafficking have been identified, and emerging evidence supports the importance of subcellular compartmentalization of signaling components at the right time and in the right place. In addition, the translocation of GLUT4 in adipocytes requires insulin stimulation of dynamic actin remodeling at the inner surface of the plasma membrane (cortical actin) and in the perinuclear region. This results from at least two independent insulin receptor signals, one leading to the activation of phosphatidylinositol (PI) 3-kinase and the other to the activation of the Rho family small GTP-binding protein TC10. Thus, both spatial and temporal regulations of actin dynamics, both beneath the plasma membrane and around endomembranes, by insulin receptor signals are also involved in the process of GLUT4 translocation. PMID- 16702777 TI - Age-dependent differences in the time course of postural control during sensory perturbations. AB - BACKGROUND AND AIMS: With research demonstrating that older adults are more likely to sustain a loss of balance than younger adults, the need for an account of age-related differences in postural control is apparent. Several measures of balance reported in the literature have assessed balance using an average or summative measure over the course of a trial, typically lasting several seconds. One limitation related to these measures is the inability to assess the time course of postural control occurring throughout the trial. To this end, the current investigation assessed the temporal changes in balance both as a function of age and sensory environment. METHODS: Postural control was assessed from 10 older adults (69.3+/-4.7 years) and 10 younger adults (22.1+/-1.7 years) over the course of fifteen seconds in three different sensory environments. The sensory manipulations involved sway-referenced rotation of the floor and/or the visual surround. RESULTS: Significant differences (p=0.001) in overall postural stability were observed between the two groups on all three sensory conditions. Additionally, time-course changes were observed between the two groups when the environment did not cause sensory conflict (sway-referenced floor or room only). However, when the environment created a situation of sensory conflict (sway referenced floor and room) both groups followed the same time-course of postural changes. CONCLUSIONS: Interestingly, the time course of postural control for the older adults on the easiest condition was mimicked by the younger adults on a more challenging condition. However, when faced with sensory conflict there were no group differences in the time course of postural control. The findings suggest an age-related decline in the temporal control of posture in altered sensory environments. PMID- 16702776 TI - Muscle training for bone strength. AB - The main function of bone is to provide the mechanical integrity for locomotion and protection; accordingly, bone mass and architecture are adjusted to control the strains produced by mechanical load and muscular activity. Age-related patterns involve peak bone mass during growth, a plateau in adulthood, and bone loss during aging. The decline in bone mass and structural integrity results in increased risk of fractures, particularly in post-menopausal women. Athletes competing in strength and power events, such as weight-lifting and jumping, have superior bone mass and structure compared with their untrained counterparts in all age groups. Exercise seems to be most effective during rapid growth, the average gain in bone mineral content (BMC) and density (BMD) in controlled trials being of the order of 2-5% per year. The net gain of BMD after exercise interventions among older people is modest, at a level of 1-3% per year, but it is not clear whether positive effects can be maintained over a longer time. Although aerobic exercise is important in maintaining overall health, the resistance type of muscle training may be more applicable to the basic rules of bone adaptation and site-specific effects of exercise, have more favorable effects in maintaining or improving bone mass and architecture, and be safe and feasible for older people. It has been suggested that there is an opportunity for resistance training, for improved effects on BMD in postmenopausal women in bones which have less daily loading. In addition to BMC and BMD, bone geometry and mass distribution may also change as a result of training and other treatment, such as hormonal replacement therapy, thereby further improving bone strength and reducing fracture risk. Appropriate training regimens may reduce the risk of falls and the severity of fall-related injuries, and also constitute potential therapy to improve functional ability and the quality of life in osteoporotic patients. However, further research is needed on dose-response relationships between exercise and bone strength, the feasibility of high-load, high-speed and impact-type of physical training, and the risks and benefits of intensive exercisein elderly individuals. PMID- 16702778 TI - Concordance and discordance between two measures of lower extremity function: 400 meter self-paced walk and SPPB. AB - BACKGROUND AND AIMS: The purpose of the study was to assess the concurrent validity of the 400 meter self-paced walk test (400-m W) against the commonly used short physical performance battery (SPPB). A secondary purpose was to determine whether the 400-m W could better discriminate physical performance among high functioning older adults by examining the distribution of 400-m W scores. METHODS: 101 men and women (80.8+/-0.4 years) were recruited to participate in the study. The 400-mW and SPPB assessed lower extremity function. Lower extremity muscle strength, power, and contraction velocity was assessed using bilateral leg press (LP). Health history was obtained with questionnaire. RESULTS: 400-m W demonstrated moderate correlations with SPPB (Pearson r=0.74; p< 0.001). In 36 high functioning individuals (SPPB score= 10, 11, 12), participants above the median 400-m W performance time (n=20) had more medical conditions (2.8+/-0.4 vs 1.7+/-0.3; p=0.038), more reported falls (0.80+/-0.2 vs 0.19+/-0.1; p=0.016), more medications (3.7+/-0.4 vs 1.8+/-0.4; p=0.001), had lower LP power at 70% of the one repetition maximum (1RM) (336+/-45 W vs 663+/-78 W; p=0.001) and 40% 1RM (329+/-43 W vs 580+/-75 W; p=0.005), and had slower LP contraction velocity at 40% 1RM (77+/-5.5 m/s vs 112+/-8.4 m/s; p=0.001) compared with those below the median (n=16). CONCLUSIONS: A substantial number of apparently well functioning older adults demonstrated some limitations in the ability to walk 400 meters. Use of the 400-m W may be justified to obtain information to better discriminate among high functioning elderly. PMID- 16702779 TI - Ankle force and rate of force production increase following high intensity strength training in frail older adults. AB - BACKGROUND AND AIMS: The most common cause of accidental injury and death in people over age 65 results from impacts associated with falling. Balance impaired older adults have poorer balance control than healthy young adults or healthy older adults. Lack of sufficient lower extremity strength and inability to rapidly produce muscle force may contribute to diminished balance control in the elderly. This study evaluated the effect of a 10-week high intensity strength training program targeting key lower extremity muscles for the purpose of improving postural control in frail older adults. METHODS: Thirteen experimental and fourteen control subjects, all balance impaired older adults were evaluated in response to unexpected platform perturbations that simulated slips. RESULTS: Following strength training the experimental group was significantly stronger than the control group. Mean ankle moments improved in the experimental group following strength training during forward sway (Right: p=0.067, Left: p=0.009) and backward sway (Right: p=0.031, Left: p=0.058). For the backward sway condition the ankle rate of torque production increased significantly in the experimental group (Right: p=0.016, Left: p=0.031). CONCLUSIONS: Enhancement of lower extremity strength contributed to improvements in balance stability demonstrated by greater ankle force production, in response to balance threats. PMID- 16702781 TI - Deep neck infection in elderly patients. A single institution experience (2000 2004). AB - BACKGROUND AND AIMS: Immunosenescence, the age-related decline in immunologic function in healthy individuals, seems to contribute to increased susceptibility to bacterial infections in the elderly population. The present study describes elderly patients' susceptibility to deep neck infection and prognosis. METHODS: Between January 2000 and March 2004, 103 patients were admitted to the Department of Otolaryngology, University of Padova for deep neck infection. Twenty-four patients (23%) were over 65 (elderly patients). The remaining 79 patients (77%) aged < or =65 years (adult non-elderly patients) were also studied. Presentation modalities, origin of infection, site of deep neck infection, radiological investigations, bacteriology, treatment and outcome were all studied. RESULTS: Hypertension and diabetes mellitus were the most commonly associated systemic diseases in both elderly and non-elderly patients. The number of patients with associated systemic diseases was significantly higher in the elderly group. The most common cause of deep neck infection was dental infection in both age groups. In the elderly group, salivary gland origin had the same incidence as dental origin. Twenty-two patients (6 elderly patients) were treated only with intravenous antibiotic therapy and intravenous steroids. Overall, in 81 cases (78.6%) (18 elderly patients) medical plus surgical procedures were indicated. None of the treated patients died of deep neck infection or its complications. CONCLUSIONS: Although the incidence of associated systemic diseases and complications of deep neck infections were higher in the elderly group, our medical or medical plus surgical approaches to deep neck infections, based on clinical and radiological evidence, were successful in all patients treated. PMID- 16702780 TI - Health status in the oldest old. Age and sex differences in the Umea 85+ Study. AB - BACKGROUND AND AIMS: With an increasing population aged 85 years and over, the aim of this study was to describe health status and living conditions in the oldest old and to estimate age and sex differences in a Northern European population. METHODS: A population-based cross-sectional study, The Umea 85+ Study, was carried out in the municipality of Umea in northern Sweden. Out of 319 eligible participants aged 85, 90 and 95 years and over, 253 participated. Structured interviews and assessments were conducted with the participants in their homes, and data were also collected from relatives, caregivers and medical charts. Cognition was screened with the Mini-Mental State Examination (MMSE), depressive symptoms with the Geriatric Depression Scale-15 (GDS-15) and nutritional status with the Mini Nutritional Assessment (MNA). Activities of daily living (ADL) were assessed applying the Staircase of ADL (including Katz' Index of ADL) and morale with the Philadelphia Geriatric Center Morale Scale (PGCMS). Participants also rated their own health. RESULTS: Over half of the participants had hypertension, one out of four was depressed, and the same proportion had had a hip fracture; the mean number of drugs taken was 6.4+/-4.0. Younger participants had lower rates of diagnoses and prescribed drugs, and were less dependent in ADL and other functional variables; men had lower rates of diagnoses and reported symptoms. The majority of participants rated their general health and morale as good. CONCLUSIONS: There were large variations in social, medical and functional variables within and between age and sex groups. This northern population of the oldest old seems to have a very high prevalence of hypertension, depression, hip fractures, and many prescribed drugs. PMID- 16702783 TI - Cognitive rehabilitation in Alzheimer's Disease. AB - Non-pharmacological treatment in Alzheimer's Disease has gained great attention in recent years. The limited efficacy of drug therapy and the plasticity of human central nervous system are the two main reasons that explain this growing interest in rehabilitation. Different approaches have been developed. Here we discuss the efficacy of non-pharmacological therapy in the frame of two main approaches: Multistrategy Approaches (Reality Orientation, Reminiscence Therapy and Validation Therapy) and Cognitive Methods. PMID- 16702782 TI - Classification of residents in nursing homes in Tuscany (Italy) using Resource Utilization Groups Version III (RUG-III). AB - BACKGROUND AND AIMS: Samples of nursing homes in Tuscany (Italy) classify their residents and determine their case-mix according to the Resource Utilization Groups System, Version III (RUG-III). METHODS: A large sample of nursing homes was selected, based on willingness to participate, representation of both public and private institutions, and wide geographic representation. Two registered nurses assessed all residents using the RUG questionnaire. The information collected was then used to group residents into 44 RUGs, and facility-specific case-mix indices were calculated using the RUG-specific weights previously validated in Italy. RESULTS: A total of 3981 residents from 93 nursing homes were assessed. Most residents were over 75 years old (87.4%) and women (68.6%). A large percentage was classified into RUGs within the following primary categories: reduced physical function (33.6%), impaired cognition (17.6%) and clinically complex (17.6%). The resulting nursing home case-mix indices ranged from 0.627 to 1.108 (mean 0.807+/-0.110). No significant association was found between type of facility, level of fees, or extent of staff in the nursing homes and their case-mix indices. CONCLUSIONS: RUGIII can provide information on types of nursing home residents and their care needs. This is useful for monitoring and evaluating long-term care services for the elderly, and allows for more effective planning and allocation of staffing and financial resources. PMID- 16702784 TI - Cerebrovascular disease in the elderly: lipoprotein metabolism and cognitive decline. AB - Data concerning the treatment of lipoprotein disturbances in patients with cerebrovascular disease (CVD) are less robust than those for coronary heart disease (CHD), raising clinical questions as to which is the appropriate therapeutic approach to stroke patients. Although observational cohort studies have failed to demonstrate an association between lipoprotein disorders and stroke incidence, recently completed trials of subjects at risk for CHD have shown that statins reduce not only the risk of myocardial infarction and death, but also that of brain infarction and transient ischemic attacks. At present, it seems reasonable to conclude that stroke patients with undesirable lipid profiles who have a history of CHD should receive specific treatment for the lipid disorder. Recommendations are more problematic for stroke patients with lipid disorder but no history of CHD. Furthermore, many of the risk factors for CVD and vascular dementia (VaD), including serum total cholesterol (TC), lipoprotein(a), diabetes, atrial fibrillation, hypertension, apolipoprotein E levels, and atherosclerosis, have also been shown to increase the risk of Alzheimer's disease (AD). In a recent study, we estimated the prevalence, incidence and rate of progression of Mild Cognitive Impairment (MCI) to dementia, and correlated vascular risk factors with incident MCI and its progression to dementia. We evaluated 2963 individuals from the population-based sample of 5632 subjects 65 84 years old of the Italian Longitudinal Study on Aging, with a 3.5-year follow up. We found a progression rate to dementia (all causes) of 3.8/100 person-years. Furthermore, age was a risk factor for incident MCI, while education was protective, and serum TC evidenced a non-significant borderline trend for a protective effect. There was a non-significant trend for stroke as a risk factor of progression of MCI to dementia. In conclusion, in our population, among MCI patients who progressed to dementia, 60% progressed to AD and 33% to VaD. Vascular risk factors and CVD may influence the development of MCI and the rate of progression to dementia. PMID- 16702786 TI - RACK-1 expression and cytokine production in leukocytes obtained from AD patients. AB - BACKGROUND AND AIMS: The purpose of this study was to evaluate in vitro cytokine production in blood leukocytes obtained from sporadic AD patients, aged controls and young individuals. METHODS: Diluted whole blood was treated in the presence or absence of LPS (1 microg/mL) for varying times (3-48 h). The release of IL-8, IL-10 and TNF-alpha in conditioned media was evaluated by commercially available sandwich ELISA. RESULTS: Data obtained are indicative of the presence of an unregulated systemic inflammation in AD patients. Leukocytes obtained from AD patients had increased spontaneous TNF-alpha release and decreased LPS-induced IL 10 production, in comparison with both old controls and young subjects, while identical IL-8 production was observed in all groups. The last finding indicates that there was no shift in the potency or efficacy of the response towards LPS with aging, but alterations in downstream signal transduction pathways are probably altered with aging and pathological conditions. CONCLUSIONS: The dysregulation of cytokine production observed in AD patients may partially be explained by a significant reduction in the expression of RACK-1, a protein crucial for integration of signaling pathways with different physiological functions, such as cytokine production. PMID- 16702785 TI - Role of acetylcholinesterase inhibitors in pharmacological regulation of amyloid precursor protein processing. AB - The triggering events leading to the selective neurodegeneration observed in Alzheimer brains are not yet completely understood. They thus create a great challenge for the definition of a resolutive treatment for the causes and symptoms of Alzheimer's Disease (AD). Since the current therapeutic option for AD patients is the use of acetylcholinesterase inhibitors (AChEIs), several authors have examined whether these drugs can also affect the expression and metabolism of the amyloid precursor protein (AbetaPP). The rationale behind these studies was based on the fact that the literature suggests that cholinergic activities are also involved in the regulation of AbetaPP metabolism. Therefore, the characterization of these aspects of AD pharmacology may allow cholinergic drugs to be tested for their ability to intervene at different levels of the pathogenetic chain, other than providing a replacement therapy for lost neurotransmitters. This paper reviews the evidence that many of these drugs, although with different qualitative effects, are able to modulate the metabolism and expression of AbetaPP. This effect is often sustained by an indirect cholinergic mechanism and does not affect the mRNA expression of the precursor, although some other authors have demonstrated an effect on post-transcriptional regulation of AbetaPP expression. In addition to the effect on AbetaPP processing, we recently explored the possibility that these molecules affect a gene expression program beyond the classical pharmacological effects, for insights on possibly unexplored pathways of intervention in AD. PMID- 16702787 TI - Pharmacological treatment of Alzheimer's Disease. AB - BACKGROUND AND AIMS: The treatment of Alzheimer's disease (AD) is a challenge for physician, families, and patients. An individualized, multimodal treatment plan addressing the treatment of cognitive, behavioural and functional decline is essential. Aim of the paper is to describe the principal components of the treatment plan of AD patients. METHODS: A review of the recent literature was performed. RESULTS: Acetylcholinesterase inhibitors (AChEIs) play an important role in the improvement of cognitive decline in mild to moderate AD, even if the improvement is not permanent. Data obtained from the CRONOS project (involving about 500 Alzheimer Evaluation Units) replicate in the real world those obtained in controlled trials, confirming that AD patients may benefit from AChEI treatment. Treatment of behavioral and psychological symptoms of dementia (BPSD) requires education of caregivers, non pharmacological interventions, identification and treatment of medical illnesses or environmental precipitating conditions, specific pharmacological treatment. Traditional neuroleptics are widely used for BPSD treatment, but limited data support their use, and side effects are frequent and severe. Atypical antipsychotics are effective in treating BPSD, and safer than traditional neuroleptics. However, the increased risk of cerebrovascular accident in patients taking risperidone or olanzapine limited currently their use in demented subjects. The use of antidepressant drugs, as well as behavioral approach, may improve depressive symptoms frequently accompanying AD. CONCLUSIONS: Although at present there is no cure for AD, several drug treatments and care strategies may improve or stabilize cognitive and behavioral symptoms, and improve the quality of life of patients and families. PMID- 16702788 TI - Immunological and immunogenetic markers in sporadic Alzheimer's disease. AB - BACKGROUND: Common polymorphisms of genes controlling inflammation-modulating cytokines and acute-phase proteins which play important roles in the pathogenesis of Alzheimer's disease (AD) have been shown to be associated with AD. AIMS: The immunological and immunogenetic markers potentially useful for the AD risk evaluation and diagnosis are briefly reviewed. CONCLUSION: The state-of-the-art of immunological and immunogenetic markers of AD indicates that new tools and strategies are necessary to identify gene products useful as diagnostic tools. PMID- 16702789 TI - Stress and dementia: the role of the hypothalamicpituitary-adrenal axis. AB - Hippocampus plays a crucial role in learning and memory and, in spite of its remarkable plasticity, it is also particularly sensitive to stress hormones due to its high concentration of corticosteroid receptors. Indeed, adrenal steroids modulate hippocampal plasticity, acting on excitability and long term potentiation or depression. By a chronobiological approach, we studied the cortisol and DHEAS secretion in clinically healthy old subjects and in age matched demented patients, including both the degenerative and the vascular type. When compared to young controls, both clinically healthy elderly subjects and demented patients, particularly those with AD, had significantly higher cortisol levels at night time, i.e. at the moment of the maximal sensitivity of HPA axis to stimulatory or inhibitory inputs. At the same time, a clear age- and disease dependent reduction of DHEAS secretion was found. Thus the cortisol to DHEAS molar ratio was significantly higher in healthy old subjects, and even more in demented patients, when compared to young controls, and significantly linked to both age and cognitive impairment. Finally, the quantitative and qualitative changes of the adrenal secretory pattern were significantly correlated with the decline of hippocampal volumes, measured by MRI. In conclusion, several lines of evidence deal with a pathogenetic role of stress hormones in the occurrence and progression of cognitive disorders in elderly subjects. The consequent hippocampal neuronal impairment may in turn be responsible for the continuous activation of HPA axis and the increased hypothalamic expression of vasopressin and corticotropin releasing hormone. PMID- 16702790 TI - T-cell dysregulation caused by chronic antigenic stress: the role of CMV in immunosenescence? AB - Dysfunctional T-cell immunity contributes to susceptibility to infectious disease in the elderly. A characteristic feature of this "immunosenescence" is the predominance of clonal expansions of CD8 cells and decreased diversity of the T cell antigen receptor repertoire. Lifelong chronic antigenic stress commonly caused by infection with persistent activating herpes viruses causes the accumulation of anergic, apoptosis-resistant CD8 T cells. These dysfunctional cells are indirectly immunosuppressive by tasking up the "immunological space" as well as directly suppressive via blockade of antigen presenting cells or cytokine secretion. They are associated with an emerging "immunological risk profile" predicting mortality in longitudinal studies of very old people. It is therefore hypothesized that for that majority of elderly people infected with cytomegalovirus (CMV), which seems to act as the dominant chronic stressor, anti viral strategies would be of benefit in abrogating some of the detrimental clinical manifestations of immunosenescence. PMID- 16702791 TI - Guidelines for clinical applications of spatio-temporal gait analysis in older adults. AB - Quantifying spatio-temporal gait parameters in stationary and ambulatory geriatric settings may aid the early identification of potential fallers, as well as the documentation of illness-specific gait disorders and intervention-related changes in rehabilitative medicine. Bringing gait analysis out of the laboratory and into a clinical setting is the goal of the European GAITRite network group, initiated in 2003 in Geneva. To enhance reproducibility of gait measures and for better comparability of outcomes in clinical environments, a consensus on data collection was formulated and presented at the 2nd European GAITRite Meeting in Marseilles. The guidelines presented here are intended to facilitate network collaborations and to provide guidance to clinicians who wish to implement spatio temporal gait analysis in clinical settings. PMID- 16702792 TI - Milk-alkali syndrome: a historical review and description of the modern version of the syndrome. AB - Formerly recognized primarily for its historic interest as a disorder found in those taking milk and bicarbonate for peptide ulcer disease, milk-alkali syndrome (MAS) is experiencing a resurgence in its incidence largely due to the increased usage of calcium carbonate. The modern version of MAS affects a different patient population and has a different etiologic basis than was characterized in the original descriptions of the syndrome. Advances in parathyroid hormone measurement have allowed for improved diagnostic separation between MAS and hyperparathyroidism and have further explained some of the physiologic responses in the resolution of hypercalcemia. We have reviewed the reasons for the increasing incidence of MAS, described the typical patient with the modern form of the syndrome, and further elaborated on the pathophysiology of MAS, as it is currently understood. MAS is an important diagnostic consideration in the patient with hypercalcemia because the syndrome is now common and prompt diagnosis limits permanent kidney function impairment but depends strongly on considering the diagnosis as well as obtaining an over-the-counter medication history. PMID- 16702793 TI - Overview of the diagnosis and management of diabetic ketoacidosis. AB - Diabetic ketoacidosis is an acute complication of diabetes mellitus that can be life-threatening if not treated properly. Once thought to occur only in patients with type 1 diabetes, diabetic ketoacidosis has been also observed in patients with type 2 diabetes under certain conditions. The basic underlying mechanism for diabetic ketoacidosis is insulin deficiency coupled with elevated levels of counter-regulatory hormones, such as glucagon, cortisol, catecholamines, and growth hormone. Diabetic ketoacidosis can be the initial presentation of diabetes mellitus or precipitated in known diabetic patients by many factors, most commonly infection. The management of diabetic ketoacidosis involves careful clinical evaluation, correction of metabolic abnormalities, identification and treatment of precipitating and comorbid conditions, appropriate long-term treatment of diabetes, and plans to prevent recurrence. Certain areas need further research, such as indications for the use of bicarbonate and phosphates and the use of intravenous rapid-acting insulin. PMID- 16702794 TI - Prevalence of active hepatitis C virus infection in patients with systemic lupus erythematosus. AB - OBJECTIVE: Hepatitis C virus (HCV) infection is associated with various autoimmune disorders and can mimic systemic lupus erythematosus (SLE) clinically and serologically. There are few reports of prevalence of HCV infection in patients with SLE. The aim of this study was to determine the prevalence of HCV viremia by polymerase chain reaction (PCR) in patients with SLE. METHODS: We tested sera from 40 consecutive patients with SLE collected from 1993 to 2000. All of the patients had HCV viral load measured by PCR. The results were compared with the prevalence of HCV viremia in a control group of blood donors in our geographic area as well as in United States general population. RESULTS: HCV was detected in 4 of 40 patients (10%). The prevalence of HCV in our area blood donors is 130 cases per 100,000 persons (0.13%; P<0.0001). The prevalence of HCV infection in the United States general population, screened by PCR, is 1330 cases per 100,000 people (1.33%; P=0.002). The prevalence of HCV infection was significantly higher in our SLE patients than in our area blood donors. The frequency of HCV infection was also higher than that of the United States general population. CONCLUSION: Our observations support those of other investigators who have reported an increased prevalence of HCV infection in SLE patients. Further detailed investigation of this association may help in understanding the pathogenesis of SLE. HCV infection should be tested when the diagnosis of SLE is considered. PMID- 16702795 TI - Addition of insulin to oral therapy in patients with type 2 diabetes. AB - BACKGROUND: A majority of individuals with type 2 diabetes will eventually require exogenous insulin therapy to achieve or maintain glycemic control. This review provides practical recommendations for adding insulin therapy for patients with type 2 diabetes whose glucose levels are inadequately controlled with oral medications. METHODS: We used a systematic review of MEDLINE to retrieve relevant articles from 1990 to 2004 using the search terms insulin therapy, combination oral therapy, glycemic control, insulin analogs, insulin glargine, and basal insulin, which we supplemented with a review of clinical practice guidelines from the American Diabetes Association and the American Association of Clinical Endocrinologists. RESULTS: Type 2 diabetes mellitus is becoming more common in the United States and is likely to increase in prevalence as obesity, a risk factor for type 2 diabetes, likewise increases. Treatment often begins with oral monotherapy, but after 3 years of treatment, more than half of patients will require more than one pharmacological agent, and eventually most patients will require insulin. Adding insulin to oral therapy at an earlier stage in treatment provides improved glycemic control without promoting increased hypoglycemia or weight gain, lowers the risk of microvascular complications by 25%, and reduces the amount of insulin patients require. Various insulin preparations, including the newer analog insulins, with different onsets and durations of action are available to help meet individual patients' dosing needs. CONCLUSIONS: The addition of insulin to oral antidiabetic therapy can improve glycemic control. Newer insulin analogs can emulate normal physiologic insulin secretion and potentially limit diabetes-related comorbidity. PMID- 16702796 TI - C-reactive protein in predicting coronary artery disease in subjects with aortic valve sclerosis before diagnostic coronary angiography. AB - BACKGROUND: Although previous studies have suggested that aortic valve sclerosis (AVS) shares common histologic features with atherosclerosis and is an indicator of significant coronary artery disease (CAD), many patients with aortic valve disease do not have coexisting coronary atherosclerotic disease and vice versa. It is important to find the subjects with AVS who are most likely to have concomitant CAD and require aggressive evaluation. HYPOTHESIS: We hypothesized that the systemic inflammatory marker, high-sensitive C-reactive protein (hs CRP), may be associated with AVS, and may be helpful before coronary angiography in identifying the presence of concomitant CAD in patients with AVS. METHODS: This study included 227 patients with suspected CAD undergoing transthoracic echocardiography and coronary angiography. AVS was defined as a focal area of increased echogenicity and thickening of the aortic valve leaflets without restriction in motion. Data of atherosclerotic risk factors including hs-CRP were collected. RESULTS: Technically satisfactory ultrasound recordings were obtained in 217 subjects (96% of enrolled patients). Patients with AVS were older (65+/-10 vs. 60+/-10 years old; P=0.0004), had higher serum creatinine levels (115.2+/ 79.7 vs. 88.6 +/-35.4 micromol/L; P=0.04), and had greater prevalence of obstructive CAD (75% vs. 53%; P=0.001) than those with normal aortic valves. CRP levels were not associated with AVS, and failed to predict concomitant CAD in patients with AVS. Additionally, none of the established risk factors were independent predictors of the presence of CAD in AVS patients. CONCLUSION: Hs-CRP levels appear to not be associated with AVS, and are of little value in terms of predicting the presence of concurrent CAD before coronary procedure. PMID- 16702797 TI - Effect of lipid levels and lipid-lowering therapy on restenosis after coronary artery stenting. AB - BACKGROUND: Recent experimental and clinical data suggest that lowering serum lipid levels with statins may prevent or delay the process of restenosis. The purpose of this trial is to determine whether lipid levels relate to restenosis and/or whether statin therapy can prevent or delay the process of restenosis after intracoronary stenting. METHODS: One hundred thirty-six patients who underwent single coronary artery stenting from June 1995 to June 1997 in our institution were included in the study. All these patients were followed for at least 9 months (mean 392+/-148 days) for major adverse cardiac events (MACE). We defined as MACE the occurrence of death, myocardial infarction, or need for target lesion revascularization. From this cohort, 103 patients had at least one lipid parameter from the lipid profile evaluated within 2 months from the date of the procedure. Patients who had the stent because of an acute myocardial infarction were included in the study only if their lipid profile was evaluated before or at least 6 weeks after the event. Patients with triglyceride levels above 500 had both triglyceride and low-density lipoprotein cholesterol levels excluded from the statistical analysis. Patients were divided into two groups based on lipid levels: normal (Group I; n=31) and elevated (Group II; n=72). Patient outcomes were also analyzed by statin therapy use. RESULTS: There was no significant difference in MACE rates between the two groups when outcomes were analyzed by lipid levels (22.6% versus 20.8% P=0.8). Furthermore, outcomes were analyzed by use of statin therapy (Group III, n=53, on statin versus Group IV, n=50, on no statin). There was also no difference in MACE rates between the two groups (20.8% versus 22%; P=0.8). CONCLUSION: The process of restenosis has unique features that differentiate it from atherosclerosis. Although lipid lowering therapy is crucial in delaying the process of atherosclerosis, its role in the prevention of restenosis is yet to be proven. PMID- 16702798 TI - Eosinophilic pancreatic infiltration as a manifestation of lung carcinoma. AB - A 68-year-old man reported upper abdominal pain during the previous 3 months that worsened in the last 2 days. He had a history of lung squamous cell carcinoma for which he underwent right lung lobectomy 3 years earlier. Preliminary blood tests showed leucocytosis with marked eosinophilia. No evidence of recurrent malignancy was detected, but computed tomography scan of the abdomen revealed an enlarged and edematous pancreas with hyperemia and infiltration of the peripancreatic fat. Fine needle aspiration from the lesion revealed inflammatory infiltration predominantly composed of eosinophils. The diagnosis of eosinophilic pancreatitis was suggested and the patient was placed on prednisone, but without any clinical or laboratory improvement. Two months later, the patient developed severe dyspnea, chylothorax, and acute renal failure. Cytologic studies of the pleural fluid revealed malignant cells from recurrent lung squamous cell carcinoma. The disease course was characterized by rapid deterioration and a fatal outcome. To the authors' knowledge, eosinophilic pancreatic infiltration as a manifestation of lung carcinoma has not been previously reported. PMID- 16702799 TI - Atypical sarcoidosis: a case report and literature review. AB - An African American man was admitted with multiple systemic symptoms. The work-up revealed a unilateral cavitary lung mass with hilar adenopathy, sub-centimeter abdominal mesenteric lymph nodes, and a natural killer cell lymphocytosis in the bone marrow. Transbronchial biopsy revealed areas of noncaseating granulomas suspicious for sarcoidosis. Additional studies for infectious and malignant agents were negative. PMID- 16702800 TI - A case of cholesterol embolism confirmed by skin biopsy and successfully treated with statins and steroids. AB - Although cholesterol embolism syndrome was recognized as a clinicopathologic entity more than 50 years ago, it is attracting growing attention recently. It is a multisystemic disorder in which cholesterol crystals released from atherosclerotic plaques obstruct small arterioles, resulting in local ischemia and end-organ damage. There are no established treatments, and with the limited treatment options available, it is important to make the diagnosis as early as possible. We present the case of a 68-year-old man with cholesterol embolism who had a few fluttering atheromas in the aorta, as demonstrated by transesophageal ultrasonography. The diagnosis was confirmed by skin biopsy, and treatment with statins and steroids proved effective, as renal failure progressively improved. This case emphasizes the importance of early diagnosis and shows the possible therapeutic effects of statins and steroids for patients with this syndrome. PMID- 16702801 TI - Rapid recurrence of hyperparathyroidism from both nodularly hyperplastic autograft at forearm and residual tissues at neck after parathyroidectomy in a hemodialysis patient with calciphylaxis. AB - A 42-year-old woman underwent hemodialysis secondary to diabetic nephropathy. Total parathyroidectomy with forearm autograft was performed due to secondary hyperparathyroidism (HPT) complicated with calciphylaxis. Rapidly progressive enlargement of autograft with unusual "gourd-shape" developed, and then it was removed. Pathologic examination of the autograft disclosed multinodular hyperplasia. Residual parathyroid gland in the retrothyroid region was found later. Rapidly recurrent HPT originating from both the residual parathyroid tissues and the enlarged autograft within such short time after parathyroidectomy is rare in the literature. The multinodular hyperplasia pattern of the parathyroid gland may be a major factor for such rapid recurrence. In addition to good control of calcium and phosphate, regular follow-up of parathyroid hormone level and imaging studies of not only autografted gland at the forearm but also possibly residual parathyroid tissues at the neck are important for monitoring recurrence in maintenance hemodialysis patients after parathyroidectomy with forearm autograft, especially in those with pathologic type of nodular hyperplasia and calciphylaxis. PMID- 16702802 TI - Syndrome of inappropriate antidiuretic hormone secretion in a patient with cryptococcal meningoencephalitis: a hidden mediastinal small cell carcinoma. AB - We report a 59-year-old man with cryptococcal meningoencephalitis who presented with typical features of syndrome of inappropriate antidiuretic hormone secretion (SIADH). There was no evidence of lung disease or drugs causing SIADH. Studies for endocrine functions and tumor markers were normal. The patient's hyponatremia was refractory to treatment (Na 120-125 mmol/L) after resolution of the cryptococcal infection. Three months after completion of antifungal treatment, a radiograph of the chest disclosed widening of the mediastinum. Computed tomography revealed a mediastinal mass. A tentative diagnosis of extrapulmonary cryptococcoma or a tumor was made. Positron emission tomography using F fluorodeoxy glucose demonstrated an intense uptake mass in the mediastinum without lesions elsewhere in the body. A biopsy of the mass clearly showed small cell carcinoma (SCC) with positive immunoreactivity for arginine vasopressin and atrial natriuretic peptide. The hyponatremia completely resolved and tumor size dramatically decreased after chemotherapy. To the best of our knowledge, this is the first case of mediastinal SCC associated with SIADH. Patients with otherwise unexplained SIADH should be meticulously scrutinized for a possible malignancy. PMID- 16702803 TI - Pyomyositis with hepatic and perinephric abscesses caused by Candida albicans in a diabetic nephropathy patient. AB - Both disseminated candidiasis and pyomyositis are rare and mainly encountered in severely immunocompromised hosts. To our knowledge, Candida albicans related pyomyositis with formation of multiple visceral abscesses in a diabetic nephropathy patient has never previously been reported. A 47-year-old man with diabetic nephropathy and alcoholic liver disease developed disseminated candidiasis, with the initial presentation of pyomyositis. Debridement was performed and intravenous fluconazole commenced. Despite development of a single hepatic and multiple perinephric abscesses, the patient made a full recovery after completion of a 12-week course of intravenous fluconazole therapy. Candida species should be considered a potential pathogen in patients with predisposing factors. PMID- 16702804 TI - Recent developments in targeting the mammalian target of rapamycin (mTOR) kinase pathway. AB - The mammalian target of rapamycin (mTOR) is a threonine kinase involved in intracellular pro-survival signaling. Its activation leads to progression from the G1 to S phase of the cell cycle. Constitutive activation of the mTOR-related messengers, including phosphatidylinositol 3-kinase, Akt kinase, ribosomal p70S6 kinase and eukaryotic translation initiation factor 4E-binding protein kinase, was found in numerous malignancies. Recent data indicate that the mTOR kinase pathway can be an attractive target for anti-cancer drug development. A well known mTOR inhibitor is rapamycin (RAPA), previously applied as an immunosuppressive agent in transplant studies. Recently, analogs of RAPA, such as CCI-779, RAD001 and AP23573, have been developed. All of those agents are currently being tested in patients with solid or hematological tumors in several clinical trials. This review presents recent developments in targeting the mTOR kinase pathway. PMID- 16702805 TI - N-benzyladriamycin-14-valerate (AD 198) activates protein kinase C-delta holoenzyme to trigger mitochondrial depolarization and cytochrome c release independently of permeability transition pore opening and Ca2+ influx. AB - Unlike nuclear-targeted anthracyclines, the extranuclear-targeted doxorubicin congener, N-benzyladriamycin-14-valerate (AD 198), does not interfere with normal topoisomerase II activity, but binds to the C1b regulatory domain of conventional and novel isoforms of protein kinase C (PKC). The resulting interaction leads to enzyme activation and rapid apoptosis in a variety of mammalian cell lines through a pathway involving mitochondrial events such as membrane depolarization (Deltapsim) and cytochrome c release. Unlike other triggers of apoptosis, AD 198 mediated apoptosis is unimpeded by the expression of Bcl-2 and Bcl-XL. We have further examined AD 198-induced apoptosis in 32D.3 mouse myeloid cells to determine how the anti-apoptotic effects of Bcl-2 are circumvented. The PKC-delta inhibitor, rottlerin, and transfection with a transdominant-negative PKC-delta expression vector both inhibit AD 198 cytotoxicity through inhibition of Deltapsim and cytochrome c release. While the pan-caspase inhibitor Z-VAD-FMK blocks AD 198-induced PKC-delta cleavage, however, it does not inhibit Deltapsim and cytochrome c release, indicating that AD 198 induces PKC-delta holoenzyme activation to achieve apoptotic mitochondrial effects. AD 198-mediated Deltapsim and cytochrome c release are also unaffected by cellular treatment with either the mitochondrial permeability transition pore complex (PTPC) inhibitor cyclosporin A or the Ca chelators EGTA and BAPTA-AM. These results suggest that AD 198 activates PKC-delta holoenzyme, resulting in Deltapsim and cytochrome c release through a mechanism that is independent of both PTPC activation and Ca flux across the mitochondria. PTPC-independent mitochondrial activation by AD 198 is consistent with the inability of Bcl-2 and Bcl-XL expression to block AD 198 induced apoptosis. PMID- 16702806 TI - Telomerase activity is not a key determinant of sensitivity to standard cytotoxic drugs in human esophageal carcinoma cell lines. AB - The aim of the present study was to investigate if basal telomerase activity levels may predict sensitivity to cytotoxic drugs in a panel of human esophageal carcinoma cell lines. The TRAPeze telomerase detection assay was used to investigate telomerase activity in the cell lines. Cytotoxic drug sensitivity for 20 standard cytotoxic agents was assessed using the fluorometric microculture cytotoxicity assay (FMCA). Telomerase activity was detected in all cell lines with a broad range of activity levels. Drug sensitivity also varied considerably between the cell lines. Except for a P value towards a correlation between mitoxantrone and telomerase activity (P=0.054), no statistically significant correlation was found between telomerase activity levels and sensitivity to investigated drugs, including key drugs such as cisplatin (P=0.9), 5-fluorouracil (P=0.8) and doxorubicin (P=0.54). We therefore conclude that basal telomerase activity level is not a key determinant of sensitivity to standard cytotoxic drugs in esophageal carcinoma cell lines. PMID- 16702807 TI - Structure-anti-leukemic activity relationship study of B- and D-ring modified and non-modified steroidal esters of chlorambucil. AB - In order to study the role of the steroidal moiety on the expression of anti leukemic activity, we synthesized six derivatives of chlorambucil (CHL), and tested them on leukemias P388 and L1210 in vivo and in normal human lymphocytes in vitro. Five of the six tested compounds produced submultiple toxicity, while the measured anti-leukemic potency was significantly increased. The lactamization of the B-steroidal ring rendered the molecules more potent, but the corresponding 7-oxidized derivatives proved better in both leukemias tested. The lactamization of the D-steroidal ring afforded potent compounds, regardless of the configuration of the B-ring. The best among all derivatives contains both chemical modifications and is intended as a promising key molecule that must be further studied. We speculate that in leukemic cells a tumor-specific protein is overexpressed, the steroid has the ability to bind and block this protein from carrying out its normal function, and the drug-protein complex prevents the repair of the adducts. The synthesis, physicochemical and spectroscopic data of these compounds and a modified route for the synthesis of CHL are also reported. PMID- 16702808 TI - N-methylformamide and 9-hydroxystearic acid: two anti-proliferative and differentiating agents with different modes of action in colon cancer cells. AB - N-methylformamide (NMF) is an anti-proliferative, differentiating agent studied in several cell lines as well as in preclinical and clinical trials, whose mechanisms of action are still unclear. 9-Hydroxystearic acid (9-HSA) is an endogenous product of lipid peroxidation recently identified as a new histone deacetylase 1 inhibitor. Both agents show the same anti-proliferative effects by arresting colon cancer cell growth in G0/G1. We addressed two questions. (i) Do they act by regulating G0/G1 checkpoint proteins? (ii) Does 9-HSA have differentiating effects comparable to those of NMF? The effects of NMF and 9-HSA on growth, differentiation and invasiveness of HT29, a colon cancer cell line, have been compared by using immunoprecipitation analysis, confocal microscopy, enzyme assays and invasiveness tests. The results show that the G1 arrest caused by NMF is a cell cycle exit due to p27 induction, whereas 9-HSA has no effect on the induction of this inhibitor. Evidence is presented that the arrest in early G0/G1 induced by 9-HSA is associated with the conversion of HT29 characteristics to those of a more benign phenotype, whereas the arrest in the late G1 in response to NMF is not followed by a decrease in tumorigenicity. The failure of NMF in cancer therapy indicates that both anti-proliferative and differentiating characteristics are required for an anti-tumoral agent to be effective. PMID- 16702809 TI - Cytotoxicity of a recombinant fusion protein of adenovirus early region 4 open reading frame 4 (E4orf4) and human epidermal growth factor on p53-deficient tumor cells. AB - Adenovirus early region 4 open reading frame 4 (E4orf4) protein is a novel cell death factor that selectively induces p53-independent apoptosis in cancer cells, but not in normal human cells. This study presents an approach for inhibiting p53 deficient tumor cell growth by using protein-based E4orf4 that had been genetically fused to epidermal growth factor (EGF) to ensure selective targeting of EGF receptor-overexpressing tumor cells. EGF-E4orf4 enables binding onto the cell surface and is then internalized into Saos-2 cells. The success of the process had been demonstrated by immunofluorescence assay and confocal laser microscopy. After prolonged exposure, E4orf4 remained mostly in the nuclei. EGF E4orf4 treatment of Saos-2 cells showed dose-dependent cytotoxicity. Nearly 50% of the Saos-2 cells were killed at a concentration of 250 nmol/l. In contrast, EGF-E4orf4 showed no significant inhibitory effect iresn primary cells of human umbilical vein endothelial cells. To confirm the ability of EGF-E4orf4 to induce apoptosis, DNA fragmentation was detected using BrdUTP end-labeling. Flow cytometric analysis revealed a significant increase of apoptotic cells in Saos-2 cells treated with EGF-E4orf4, but not in the case of cells cultured in plain medium (t=0.028, P<0.05). In conclusion, these preliminary results indicate that EGF-E4orf4 could show promise as a new reagent that is more efficient and less toxic in anti-cancer therapy. PMID- 16702810 TI - Thioredoxin reductase and cancer cell growth inhibition by organogold(III) compounds. AB - Thioredoxin (Trx) expression is increased in several human primary cancers associated with aggressive tumor growth and decreased patient survival, and the Trx/Trx reductase (TrxR) system therefore provides an attractive target for cancer drug development. Various gold(III) compounds with none, one, two or three carbon-gold bonds were evaluated for their capacity to inhibit TrxR and the growth of MCF-7 cancer cells in vitro. Compounds with up to two carbon-gold bonds were often potent inhibitors of TrxR with IC50 values as low as 2 nmol/l. In the presence of Trx and insulin the inhibiting capacity was much lower. However, the inhibitory concentrations of the compounds did not correlate with the ability to kill cells. Out of the organometallics tested, only compound 8 with two carbon gold bonds was able to inhibit colony formation by MCF-7 breast cancer cells at low micromolar concentrations (IC50=1.6 micromol/l). Unfortunately, the compound did not show any anti-tumor activity against MCF-7 breast cancer and HT-29 colon cancer xenografts in scid mice. PMID- 16702811 TI - Anti-cancer activity and mechanisms of 25-anhydrocimigenol-3-O-beta-D xylopyranoside isolated from Souliea vaginata on hepatomas. AB - Our previous study first revealed the cytotoxicity and relative selectivity of 25 anhydrocimigenol-3-O-beta-D-xylopyranoside (ACX) on HepG2 and R-HepG2 cells. In the present study, the anti-cancer activity and mechanisms of ACX isolated from S. vaginata were investigated both in vitro and in vivo. ACX showed significant, consistent anti-proliferative activity on hepatoma bel-7402 cells by MTT and clone formation assays with an IC50 value of 18 mumol/l. Morphological observation and flow cytometry results showed that apoptosis and G0/G1 cell cycle arrest contributed to the cytotoxic and cytostatic effects. Further studies showed that Bax and p21 protein expression were upregulated, Bcl-2 protein expression was downregulated, and poly(ADP-ribose) polymerase protein was cleaved. Moreover, ACX also exhibited a dose-dependent inhibition of tumor growth on mice implanted with H22 in vivo. These findings implicate ACX as a promising anti-cancer agent for chemotherapy of certain cancers. PMID- 16702813 TI - Effect of all-trans retinoic acid and sodium butyrate in vitro and in vivo on thyroid carcinoma xenografts. AB - Histone deacetylase (HDAC) inhibitors have been shown to reverse repression of some genes, including retinoic acid (RA) receptor beta2. In this work, we studied the effects of RA alone or combined with the HDAC inhibitor sodium butyrate (NaB) in a poorly differentiated thyroid carcinoma cell line (FTC-133) cultured in vitro or transplanted into nude mice. In vitro, the action of the xenobiotics on cell differentiation was evaluated by the measurement of alkaline phosphatase (ALP) activity. In vivo, FTC cells were injected in nude mice divided into four groups: controls, RA (1 mg/kg), NaB (50 mg/kg) in two daily injections or both RA plus NaB. Body weight, tumoral volume (TV), doubling time of the tumor, specific growth delay and inhibition of tumoral growth at day 35 were determined in each group. In vitro, RA increased the NaB-induced increase in ALP activity. In vivo, body weight and TV decreased with RA or NaB. Specific growth delay significantly increased with RA (72.5%; P < 0.001) and with NaB (31.3%; 0.02

or=160 vs. <135 mmHg) gives an estimated age sex-adjusted hazard ratio for mortality of 1.51 (95% confidence interval 1.25-1.83, P<0.001). The corresponding comparison for clinic systolic blood pressure (>or=174 vs. <148 mmHg) results in a hazard ratio of 1.02 (95% confidence interval 0.84-1.24, P=0.9). Comparing ambulatory versus clinic systolic blood pressure (10 mmHg groupings) using nested statistical modelling, removal of the ambulatory blood pressure term from the baseline Cox model (nine 10 mmHg categories) resulted in a highly significant likelihood ratio test statistic of 52.5 (df=8, P<0.0001). The corresponding result for removal of the clinic blood pressure term was 18.1 (df=8, P=0.02), thus reinforcing the finding that ambulatory blood pressure monitoring has greater prognostic significance. Ambulatory blood pressure was also a better predictor of all-cause mortality both in patients taking medication and those not taking medication at the time of monitoring. CONCLUSION: Daytime ambulatory blood pressure monitoring is a much better prognostic indicator of all cause mortality than clinic blood pressure. PMID- 16702820 TI - The choice of threshold limits for pediatric ambulatory blood pressure monitoring influences clinical decisions. AB - OBJECTIVES: Hypertension in children may be defined by blood pressure elevated above the 95th percentile according to sex and age. Population data for ambulatory blood pressure provide different age-related and sex-related threshold limits to office-derived data. We sought to determine whether, when using ambulatory blood pressure monitoring in a clinical setting, changing 95th percentile threshold limit sets from office-derived to ambulatory blood pressure derived would lead to different diagnostic decisions. METHODS: Three nephrologists who were blinded as to patient identity and limit setting method reported on 42 ambulatory blood pressure records from a mixed group of patients aged 5-18 years by using both office-derived threshold limits for the 95th centile of blood pressure and ambulatory blood pressure-derived limits. Decisions regarding the presence or absence of hypertension were compared for each patient according to the limit set. RESULTS: Thirty-five (83%) patients were considered to be hypertensive when office-derived threshold limits were used and 20% (P=0.005) fewer records were reported as showing hypertension when ambulatory blood pressure-derived threshold limits were used. When ambulatory blood pressure limits were applied, there were fewer records with an awake systolic blood pressure load >50% (P=0.004) and the average awake systolic blood pressure load was significantly lower (P<0.001). CONCLUSION: Ambulatory blood pressure normative data tend to provide higher blood pressure limits for age and sex. Consequently, when ambulatory blood pressure data are used to set threshold limits, clinical decisions based on ambulatory blood pressure may be different than when office limits are used. These findings demonstrate the importance of using the most appropriate limit sets to analyze ambulatory blood pressure and when interpreting ambulatory blood pressure-based research. PMID- 16702821 TI - Office blood pressure measurements overestimate blood pressure control in renal transplant patients. AB - OBJECTIVE: As hypertension is an important risk factor for renal allograft failure, we aimed to assess blood pressure control in renal transplant patients with deteriorating graft function using different methods of blood pressure measurements. METHODS: Forty-nine patients with a graft survival of >1 year, and with more than a two-fold increase in urinary albumin excretion, and/or an increase in serum creatinine level >20% during the previous 12 months, were included. Office blood pressure and home BP were measured, and ambulatory blood pressures were obtained in all patients. RESULTS: The mean office blood pressure (133.2+/-16.3/81.7+/-9.6 mmHg) and 24 h ambulatory blood pressure (133.1+/ 12.0/79.8+/-8.3 mmHg) were similar. Home blood pressure in the morning (144.2+/ 23.3/87.1+/-12.7 mmHg) and evening (143.2+/-20.6/86.4+/-10.3 mmHg) were significantly higher than ambulatory blood pressure (P<0.001 for both). Only 18% of the patients exhibited a reduction of >or=10% in systolic blood pressure during nighttime while 39% had an overt rise. Adequate blood pressure control was found in 53% of the patients using office blood pressure (<140/90 mmHg), contrasting 29% using home blood pressure (<135/85 mmHg), and 16% using mean 24-h ambulatory blood pressure (<125/80 mmHg). These findings were substantiated by the use of receiver-operating characteristic curve analysis. CONCLUSIONS: Using the 24-h blood pressure as a standard, home blood pressure was superior to office blood pressure in estimating blood pressure control in renal transplant patients. Nocturnal hypertension, however, was observed frequently, adding important clinical information about blood pressure control in this high-risk population. PMID- 16702822 TI - Use of 24-h ambulatory blood pressure monitoring to assess blood pressure control: a comparison of olmesartan medoxomil and amlodipine besylate. AB - OBJECTIVE: Olmesartan medoxomil is an angiotensin II receptor blocker with similar antihypertensive efficacy as the calcium channel blocker amlodipine besylate in patients with mild-to-moderate hypertension. In addition to a drug's ability to lower blood pressure, the effectiveness of the agent in enabling patients to achieve specific blood pressure targets is an important clinical consideration. This secondary analysis of a randomized, double-blind study compared the efficacy of olmesartan medoxomil with that of amlodipine besylate in achieving ambulatory blood pressure goals among patients with mild-to-moderate hypertension. METHODS: Following a 4-week placebo run-in, 440 study participants aged >or=18 years were randomized to olmesartan medoxomil (20 mg/day), amlodipine besylate (5 mg/day), or placebo for 8 weeks. The proportion of participants achieving specific systolic and diastolic ambulatory blood pressure goal levels was calculated by dividing the number of participants who had achieved a particular blood pressure goal by the total number of patients in the intent-to treat population. RESULTS: After 8 weeks of treatment, a mean 24-h ambulatory blood pressure of <130/80 or <130/85 mmHg was achieved by significantly more participants in the olmesartan medoxomil group (18.1 and 30.4%, respectively) than in the amlodipine besylate (7.0 and 14.0%, respectively) or placebo (1.9% for both) groups. The target daytime ambulatory blood pressure of <135/85 mmHg was achieved by more participants in the olmesartan medoxomil group than in the amlodipine besylate group (15.8 vs. 5.8%, respectively; P<0.01). CONCLUSION: In a previous publication of the same study, we demonstrated that starting doses of olmesartan medoxomil and amlodipine besylate produced similar mean reductions in blood pressure. In this subanalysis of the blood pressure data from that primary publication, however, olmesartan medoxomil therapy was shown to result in a greater proportion of patients achieving specific ambulatory blood pressure goals than therapy with amlodipine besylate. As blood pressure goal attainment may be of more direct clinical relevance than numerical blood pressure lowering, the achievement of blood pressure goals should be a key efficacy parameter assessed in clinical trials of antihypertensive medications. PMID- 16702823 TI - The population RDH index: a novel vector index and graphical method for statistical assessment of antihypertensive treatment reduction, duration, and homogeneity. AB - Current indices used in the evaluation of antihypertensive treatment duration and homogeneity such as the trough-peak, smoothness index, and normalized smoothness index were designed to be applied to ambulatory blood pressure monitoring recordings from individual participants. Evaluation of antihypertensive treatment in populations is often carried out by calculating these individual indices for each of the participants and providing summarizing statistics about the population, such as the mean and median. We describe a new population vector index and graphical method for the statistical assessment of antihypertensive treatment reduction, duration, and homogeneity (RDH) from ambulatory blood pressure monitoring. The population (RDH) was specifically designed as a tool to evaluate and compare blood pressure coverage offered by antihypertensive drugs over 24 h in populations. The population RDH is a three-component vector index that incorporates information about the reduction, duration, and homogeneity of antihypertensive treatment, as well as their statistical significance over the 24 h period. In addition to defining the RDH index, in this paper we also demonstrate its usefulness and advantages as an index and graphical method for antihypertensive treatment duration and homogeneity assessment by using it to analyze two data sets. PMID- 16702824 TI - Validation of the Microlife BPA100 Plus device for self-home blood pressure measurement according to the International Protocol. AB - OBJECTIVE: Current guidelines recommend that only validated devices for blood pressure measurement should be used. This study presents the validation results of the Microlife BPA100 Plus monitor for self-home blood pressure measurement, according to the European Society of Hypertension International Protocol. METHODS: Fifteen study participants were included in phase 1 and an additional 18 in phase 2 (total 33). Simultaneous blood pressure measurements were taken by two trained observers (Y tube connected mercury sphygmomanometers) four times, sequentially with three measurements taken using the tested device. Absolute differences between observer and device blood pressure were classified into three zones (within 5, 10 and 15 mmHg). The number of readings with a difference within 5 mmHg was calculated for each individual. RESULTS: In phase 1, the device produced 32, 42 and 43 measurements within 5, 10 and 15 mmHg, respectively, for systolic blood pressure and 31, 45 and 45 for diastolic blood pressure. In phase 2.1, the device produced 71, 87 and 96 measurements within 5, 10 and 15 mmHg, respectively, for systolic blood pressure, and 71, 98 and 99 measurements for diastolic blood pressure. In phase 2.2, 26 participants had at least two of their differences within 5 mmHg and three participants had no differences within 5 mmHg for systolic blood pressure, whereas for diastolic blood pressure 24 and three participants, respectively. Mean systolic blood pressure differences were -2.0+/ 6.0 mmHg and diastolic blood pressure differences were -3.1+/-4.1 mmHg. CONCLUSIONS: The Microlife BPA100 Plus device for self-home blood pressure measurement comfortably passes the validation requirements of the International Protocol and therefore can be recommended for clinical use in the adult population. PMID- 16702825 TI - Validation of a non-mercury digital auscultatory device with manual pressure registration (PMS Mandaus). AB - OBJECTIVE: To assess the clinical accuracy of a non-mercury digital auscultatory device (PMS Mandaus) with manual pressure registration in an adult population. METHODS: The accuracy of the device was assessed by predefined criteria (European Society of Hypertension protocol) in 33 study participants recruited from among patients and staff in a teaching hospital. A series of same-arm sequential blood pressure measurements were taken: first, two observers taking simultaneous mercury readings, followed by a reading with the device. A total of seven readings were taken from each participant in the sitting position. The data were then analysed according to the European Society of Hypertension protocol and the criteria of the Association for the Advancement of Medical Instrumentation. RESULTS: The device fulfilled the criteria of the European Society of Hypertension protocol, achieving a pass for both systolic and diastolic pressure. The device also passed the Association for the Advancement of Medical Instrumentation standard (the mean to be within 5+/-8 mmHg) with the mean differences being -3.2+/-3.8 mmHg for systolic blood pressure and -1.8+/-2.9 mmHg for diastolic blood pressure. CONCLUSION: The device performed in a satisfactory manner according to the European Society of Hypertension and the Association for the Advancement of Medical Instrumentation criteria. The systematic under recording related to the delay of manual pressure registration is not sufficient to preclude clinical use. PMID- 16702826 TI - Validation of two automatic devices for self-measurement of blood pressure according to the International Protocol of the European Society of Hypertension: the Omron M6 (HEM-7001-E) and the Omron R7 (HEM 637-IT). AB - BACKGROUND: Two electronic devices for self-measurement of blood pressure - a brachial monitor, the Omron M6, and a wrist monitor, the Omron R7 - were evaluated in two separate studies according to the International Protocol of the European Society of Hypertension. DESIGN: The International Validation Protocol is divided into two phases: the first phase is performed on 15 selected participants (45 pairs of blood pressure measurements); if the device passes this phase, 18 supplementary participants are included (54 pairs of blood pressure measurements) making a total number of 33 participants (99 pairs of blood pressure measurements) on whom the final validation is performed. METHODS: The same methodology recommended by the European Society of Hypertension protocol was applied for both studies. In each study and for each participant, four blood pressure measurements were taken simultaneously by two trained observers using mercury sphygmomanometers alternately with three measurements taken by the tested device. The difference between the blood pressure value given by the device and that obtained by the two observers (mean of the two observers) was calculated for each measure. The 99 pairs of blood pressure differences were classified into three categories ( or =10) previously treated with conventional antipsychotics. Patients were randomly assigned to treatment with an initial dose of olanzapine 10 mg/d or more (n = 120) or risperidone 3 mg/d or more (n = 115). The primary efficacy measure was the SANS summary score. Secondary efficacy measures included Scale for the Assessment of Positive Symptoms, Clinical Global Impression of Severity Scale, Calgary Depression Scale, and Social Functioning Scale. The response rate was defined as 30% or more of improvement in the SANS summary score. RESULTS: The mean dose throughout the study was 12.2 mg/d (+/-5.8 mg/d) for olanzapine and 4.9 mg/d (+/-2.0 mg/d) for risperidone. At 1 year, olanzapine patients showed significantly higher improvement than risperidone patients on the SANS summary (P = 0.015) and on the affective flattening (P = 0.007) and avolition/apathy (P = 0.028) SANS subscales. There were also significant improvements in favor of olanzapine in the Scale for the Assessment of Positive Symptoms summary (P = 0.021), Clinical Global Impression of Severity (P = 0.008), and Social Functioning Scale total (P < 0.001) scores. The response rate was greater (P = 0.001) in the olanzapine cohort (69.2%) than in the risperidone cohort (48.7%). Olanzapine patients reported less extrapyramidal side effects but a higher incidence of clinically important body weight increase than risperidone patients. CONCLUSIONS: Long-term treatment with olanzapine was associated with significantly better improvement in negative symptoms as compared with risperidone-treated schizophrenic outpatients with prominent negative symptoms. PMID- 16702889 TI - Treatment with venlafaxine extended release after SSRI nonresponse or intolerance: a randomized comparison of standard- and higher-dosing strategies. AB - OBJECTIVE: Evaluate efficacy of standard and higher doses of venlafaxine extended release (ER) in depressed outpatients who had either not responded to or could not tolerate an adequate trial of therapy with a selective serotonin reuptake inhibitor (SSRI). METHODS: Outpatients (n = 232) with major depressive disorder were randomly assigned to 8 weeks of treatment with either "standard" (n = 119; mean dose = 148 mg/d) or "higher" (n = 113; mean dose = 309 mg/d) dosage therapies. Between weeks 8 and 12, nonresponders in the standard dose group could receive higher dose therapy. RESULTS: Response rates in the higher dose group were significantly greater at week 8 on the Clinical Global Impressions Improvement scale (68% vs 52%; P < 0.001) and Patient Global Impressions scale (intent-to- treat; 68% vs 52%; P < 0.001). The dosing strategies did not, however, differ significantly in change in HAM-D21 total score or HAM-D21 response or remission rates. At week 12, there were no significant efficacy differences between the two groups in the intent-to-treat sample. Five side effects (constipation, sweating, hypertension, agitation, and urinary frequency) were more common in the high-dose group. CONCLUSIONS: Higher dose therapy with venlafaxine ER (ie, 300-375 mg/d) resulted in a more rapid response on some measures, but was not as well tolerated as therapy at standard doses. Although these data provide further evidence of a dose-response relationship for venlafaxine therapy results suggest that slower titration to higher doses of venlafaxine ER may improve tolerability without greatly diminishing the probability of success. PMID- 16702891 TI - Evaluation of the abuse potential of pagoclone, a partial GABAA agonist. AB - This study assessed the abuse potential of pagoclone, a partial agonist at the gamma-aminobutyric acid type A (GABAA) benzodiazepine receptor site, in healthy recreational drug users. Twenty-three young adults, who reported past recreational use of sedative drugs or alcohol, participated in 4 sessions during which capsules containing pagoclone (doses: 1.2 mg, the higher end of the proposed therapeutic dose range, and 4.8 mg, a 4-fold higher dose), diazepam (dose, 30 mg), or placebo were randomly administered under double-blind conditions. Subjective ratings of mood, drug effects, and psychomotor tests were completed at regular intervals after ingesting the capsules. On most of the standardized measures of abuse potential, pagoclone (dose, 4.8 mg) was rated as being similar to diazepam. Both drugs increased the ratings of good effects and drug liking. However, pagoclone also produced some adverse mood effects that might limit its potential to be used recreationally, and it produced fewer sedativelike effects on some measures. In general, the results with these doses indicate that the abuse potential of pagoclone is similar to that of diazepam, although its profile as a partial agonist suggests that differences between the drugs may emerge at higher doses. PMID- 16702890 TI - Venlafaxine extended release in posttraumatic stress disorder: a sertraline- and placebo-controlled study. AB - This 12-week, double-blind, multicenter trial evaluated the efficacy of venlafaxine extended release (ER), sertraline, and placebo in adult outpatients (N = 538) with a primary diagnosis of posttraumatic stress disorder (PTSD), as defined in the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, symptoms for 6 months or more and 17-item Clinician-administered PTSD Scale (CAPS-SX17) score of 60 or more. Patients were randomly assigned to receive placebo or flexible doses of venlafaxine ER (37.5-300 mg/d) or sertraline (25-200 mg/d) for 12 weeks or less. The primary outcome was the baseline-to-end point change in total CAPS-SX17 score (last observation carried forward). Secondary measures included CAPS-SX17 symptom cluster scores for reexperiencing/intrusion, avoidance/numbing, and hyperarousal; frequency of remission (CAPS-SX17 < or =20); and changes in Davidson Trauma Scale total score and symptom cluster scores for avoidance/numbing, hyperarousal, and reexperiencing/intrusion. Mean changes in CAPS-SX17 scores were -41.8, -39.4, and -33.9 for venlafaxine ER (P < 0.05 vs. placebo), sertraline, and placebo, respectively. Mean changes for venlafaxine ER, sertraline, and placebo in CAPS-SX17 cluster scores were -13.0, -11.7, and -11.0 for reexperiencing; -17.1, -16.8, and -13.7 (P < 0.05 both active treatments vs. placebo) for avoidance/numbing; and -11.8, -10.9, and -9.2 (P < 0.05 venlafaxine vs. placebo) for hyperarousal. Week 12 remission rates were venlafaxine ER 30.2% (P < 0.05 vs. placebo), sertraline 24.3%, and placebo 19.6%. The venlafaxine ER group had significantly better Davidson Trauma Scale total and cluster scores than placebo. Mean maximum daily doses were 225-mg venlafaxine ER and 151-mg sertraline. Both treatments were generally well tolerated. Study results suggest that venlafaxine ER is effective and well tolerated in the short-term treatment of PTSD. PMID- 16702892 TI - Interactions on mixing diazepam with methadone or buprenorphine in maintenance patients. AB - Benzodiazepine use by patients in methadone and buprenorphine substitution treatment is common, despite safety concerns regarding these drug interactions. The relative safety of diazepam use by methadone- or buprenorphine-treated patients has not been systematically examined. This study aimed to examine the effect of single diazepam doses, within normal therapeutic range (doses: 0, 10, and 20 mg), upon physiological, subjective, and performance measures in stable methadone and buprenorphine-treated patients. In a double-blind, randomized crossover design, methadone- or buprenorphine-treated patients were administered their normal opioid dose and either placebo, 10-, or 20-mg diazepam, in balanced order over 3 sessions. Eight methadone- and 8 buprenorphine-prescribed patients with no concurrent benzodiazepine dependence or significant comorbidity were recruited from an outpatient addiction clinic in London. Measures were taken at baseline and for 6 hours after dosing, and included physiological responses (pulse rate, blood pressure, pupil size, respiratory rate, and peripheral pO2), subjective drug effects (Addiction Research Center Inventory subscales, visual analog scales of strength of drug effect, drug-liking, and sedation), and performance measures (simple reaction time, cancellation task, digit symbol substitution task, and balance). The 10- and 20-mg diazepam doses resulted in comparable subjective experiences of greater sedation and strength of drug effects in both patient groups, and had minimal impact on physiological parameters. However, diazepam had greater peak effects on performance measures (simple reaction time, digit symbol substitution task, and cancellation time) in methadone-treated than in buprenorphine-treated patients. Diazepam may significantly alter the response to opioid substitution treatment with methadone or buprenorphine. PMID- 16702893 TI - A double-blind, placebo- and flurazepam-controlled investigation of the residual psychomotor and cognitive effects of modified release zolpidem in young healthy volunteers. AB - Short-acting hypnotic drugs, such as zolpidem, have minimal residual effects but may not provide optimal efficacy throughout the night for all insomnia patients. A modified-release formulation of zolpidem, zolpidem-MR, has been developed to overcome this limitation. This was a phase I, double-blind, 3-way crossover, placebo-controlled study to investigate the residual psychomotor and cognitive effects of a single oral dose of zolpidem-MR 12.5 mg in 18 healthy young adults. Flurazepam 30 mg was used as a positive control. No comparison with standard immediate-release zolpidem was made. Five neuropsychological tests and 2 subjective tests were performed 8 hours after dosing. The safety of zolpidem-MR was also investigated. Performance on the Critical Flicker Fusion Frequency test, Choice Reaction Time, Immediate and Delayed Word Recall, and the Compensatory Tracking Task was significantly impaired by flurazepam but not by zolpidem-MR (with the exception of the Compensatory Tracking Task) or placebo. No significant effects were observed on the Digit Symbol Substitution Test. The Leeds Sleep Evaluation Questionnaire showed that both drugs improved the ease of getting to sleep and perceived quality of sleep, whereas only flurazepam significantly impaired the ease of awakening. Neither drug scored significantly better than placebo on the Bond and Lader contentedness scale, but both induced a significant difference in calmness; only flurazepam significantly reduced alertness. The safety profile of zolpidem-MR was comparable to placebo. In conclusion, the study showed the good tolerance of zolpidem-MR in terms of residual neuropsychological effects as well as a beneficial effect on sleep quality in young healthy adults. PMID- 16702894 TI - The status of disulfiram: a half of a century later. AB - For more than 55 years, disulfiram has been approved by the Food and Drug Administration for the treatment of alcohol dependence. It is a unique medication that relies on "psychological threat" to avoid disulfiram-ethanol reactions. This paper reviews the history of disulfiram treatment, the current status of disulfiram treatment, the ensuing developments in disulfiram use in treating various addictions, and future directions. Clinical trials using disulfiram for the treatment of alcohol, cocaine, or co-occurring alcohol + cocaine dependence were included in this review. Disulfiram efficacy studies focusing on supervised, implant, and combination pharmacotherapies were also examined. In clinical trials, disulfiram has demonstrated inconsistent results in helping patients to abstain from alcohol, and patients poorly adhere to a disulfiram-treatment regimen. This has raised questions about disulfiram's practicality in the treatment of alcohol dependence. Recently, however, disulfiram has gained attention as a complementary agent to newer pharmacological medications, such as an opiate antagonist that specifically reduces alcohol craving. One hypothesis is that disulfiram would assist patients in gaining psychological control over drinking when given in conjunction with an opiate antagonist that would act directly on reducing alcohol craving. Preliminary evidence also suggests that disulfiram treatment could be a viable treatment for cocaine dependence because it was shown to reduce cocaine use among nonalcoholic, cocaine-dependent patients. PMID- 16702895 TI - To what extent do the PANSS and CGI-S overlap? AB - BACKGROUND: The Positive and Negative Symptom Scale (PANSS) and Clinical Global Impression (CGI-S) item for severity are used together to measure severity of psychotic illness. PANSS is the "gold standard" measure of efficacy, but it is not always feasible to use, yet the CGI-S requires validation. OBJECTIVES: To examine the overlap between PANSS and CGI-S. METHODS: The overlap of the PANSS and CGI-S were examined using data from 7 large antipsychotic clinical trials (n = 4287). RESULTS: Regression analysis identified 21% to 60% overlap of the measures depending on the trial and measurement point. The pooled study mean PANSS value corresponding with a CGI-S of 2, 3, and 4 were 67.1 (n = 799), 79.6 (n = 1645), and 92.4 (n = 1056), respectively. A decrease of 1 on the CGI-S corresponded to a 20% decline on the PANSS. Of the 37 planned comparisons in these studies, there was an agreement between the PANSS and CGI-S on change from baseline to end point on 32 comparisons and on dichotomized change variables (PANSS > or =-20% and CGI-S > or =-1 point) on 31 comparisons. The differences in the remaining comparisons would not have changed the conclusions of the studies. The positive and disorganized PANSS scales were the most closely related to the CGI-S, followed by hostility and negative scale with almost no association with anxiety/depression. CONCLUSIONS: The CGI-S and PANSS are correlated but are not synonymous. Both measures, however, show substantial agreement in detecting change, and the CGI-S shows overlap with the core symptoms of schizophrenia. PMID- 16702896 TI - Evaluating rater competency for CNS clinical trials. AB - Clinical trials rely on ratings accuracy to document a beneficial drug effect. This study examined rater competency with clinical nervous system rating instruments relative to previous clinical experience and participation in specific rater training programs. One thousand two hundred forty-one raters scored videotaped interviews of the Hamilton Anxiety Scale (HAM-A), Hamilton Depression Scale (HAM-D), and Young Mania Rating Scale (YMRS) during rater training programs conducted at 9 different investigator meetings. Scoring deviations relative to established acceptable scores were used to evaluate individual rater competency. Rater competency was not achieved by clinical experience alone. Previous clinical experience with mood-disordered patients ranged from none at all (18%) to 40 years in 1 rater. However, raters attending their first-ever training session (n = 485) were not differentiated on the basis of clinical experience on the HAM-A (P = 0.054), HAM-D (P = 0.06), or YMRS (P = 0.66). Alternatively, participation in repeated rater training sessions significantly improved rater competency on the HAM-A (P = 0.002), HAM-D (P < 0.001), and YMRS (P < 0.001). Furthermore, raters with clinical experience still improved with rater training. Using 5 years of clinical experience as a minimum cutoff (n = 795), raters who had participated in 5 or more training sessions significantly outperformed comparably experienced raters attending their first ever training session on the HAM-A (P = 0.003), HAM-D (P < 0.001), and YMRS (P < 0.001). The findings show that rater training improves rater competency at all levels of clinical experience. Furthermore, more stringent criteria for rater eligibility and comprehensive rater training programs can improve ratings competency. PMID- 16702897 TI - A randomized, double-blind, placebo-controlled study of citalopram in adolescents with major depressive disorder. AB - In a European, multicenter, double-blind study, 244 adolescents, 13 to 18 years old, with major depression were randomized to treatment with citalopram (n = 124) or placebo (n = 120). One third of the patients in both groups withdrew from the study. No significant differences in improvement of scores from baseline to week 12 between citalopram and placebo were found. The response rate was 59% to 61% in both groups according to the Schedule for Affective Disorders and Schizophrenia for school-aged children-Present episode version (Kiddie-SADS-P) (depression and anhedonia scores < or =2) and Montgomery Asberg Depression Rating Scale (MADRS) (> or =50% reduction). Remission (MADRS score < or =12) was achieved by 51% of patients with citalopram and 53% with placebo. A post hoc analysis revealed that more than two thirds of all patients received psychotherapy during this study. For those patients not receiving psychotherapy, there was a higher percentage of Kiddie-SADS-P responders with citalopram (41%) versus placebo (25%) and a significantly higher percentage of MADRS responders and remitters with citalopram (52% and 45%, respectively) versus placebo (22% and 19%, respectively). Mild to moderate treatment-emergent adverse events were reported in 75% citalopram and 71% of placebo patients, most commonly headache, nausea, and insomnia. Serious adverse events occurred in 14% to 15% in both groups. Suicide attempts, including suicidal thoughts and tendencies, were reported by 5 patients in the placebo group and by 14 patients in the citalopram group (not significant) with no pattern with respect to duration of treatment, time of onset, or dosage. In contrast, the suicidal ideation (Kiddie-SADS-P) single item showed worsening more frequently in the placebo (18%) than in the citalopram group (8%). PMID- 16702898 TI - Early predictors of substantial weight gain in bipolar patients treated with olanzapine. AB - To determine predictors of substantial weight gain (SWG) during treatment of bipolar disorder with olanzapine, data were pooled from 4 long-term randomized, multicenter studies in patients with bipolar mania or mixed mania (N = 948 at initiation of olanzapine). SWG was defined as gaining 5 kg or 7% of initial weight in 30 +/- 2 weeks. Logistic regression estimated odds ratios associated with early weight gain and baseline risk factors for predicting SWG. A classification system to identify patients at risk for SWG was constructed by recursive data partitioning. Baseline characteristics significantly associated with SWG included younger age, nonwhite ethnicity, lower body mass index (BMI), nonrapid cycling, and psychotic features. Weight gain of 2 or more kg in the first 3 weeks of therapy predicted SWG by 30 weeks (sensitivity = 57%; specificity = 71%). A classification system with thresholds for early weight gain, baseline BMI, and ethnicity further improved SWG predictability (sensitivity = 79%; specificity = 70%). In conclusion, patients with bipolar disorder who gained 2 to 3 kg during the first 3 weeks of treatment with olanzapine, SWG was predicted after 30 weeks of treatment. Patients with less pronounced early weight gain might still be at risk for later SWG if they have close to normal BMI (< or =27 kg/m) at treatment initiation. PMID- 16702899 TI - An examination of 26,168 Hamilton Depression Rating Scale scores administered via interactive voice response across 17 randomized clinical trials. AB - This article presents descriptive and psychometric data from 26,168 Hamilton Depression Rating Scale (HAM-D) scores administered via Interactive Voice Response (IVR) in 17 randomized clinical trials sponsored by 6 pharmaceutical companies. To provide evidence for construct validity, the IVR HAM-D scores before and after randomization are compared, and the change in the IVR HAM-D scores over time after randomization are examined. In addition, the evidence for the reliability of the IVR-administered HAM-D is presented. An examination of the distribution of first-time IVR HAM-Ds before randomization may provide useful information to researchers planning to use the IVR HAM-D as a screening tool for entry or to verify baseline severity in randomized clinical trials. PMID- 16702900 TI - Lithium poisoning: pharmacokinetics and clearance during different therapeutic measures. AB - The clinical features and pharmacokinetics of 22 lithium overdoses are described. Effectiveness of different treatment regimens regarding elimination of lithium is discussed. Origin of overdose was due to deliberate poisoning or precipitated by concomitant diseases, coadministration of drugs, or combination of both. Treatment included supportive care, diuretics (15/22), hemodialysis (HD; 9/22), and mechanical ventilation (3/22). Severity of lithium intoxication was classified in 50% as I degrees, in 41% as II degrees, and in 9% as III degrees according to Hansen and Amdisen. Renal impairment on admission was diagnosed in 82% of the patients. Half-life of lithium in serum was 3.5 +/- 0.8 hours during the first HD, and 29 +/- 14 and 29 +/- 6 hours during therapy with diuretics or supportive treatment, respectively. Lithium clearance during HD was 160 +/- 15 mL/min, and renal clearance during HD or treatment with diuretics was approximately 20 and 15 +/- 9 mL/min, respectively. Renal lithium clearance was not influenced by HD therapy. There was no difference regarding half-life and clearance between the group that had an unspecific treatment or the group treated with diuretics. Hemodialysis is the therapy of choice for emergent extracorporeal lithium elimination. Renal impairment and interaction with other drugs were the main reasons for intoxication; thus, more cautious prescription or more frequent supervision of this patient group is warranted. It seems that treatment with diuretics does not have a beneficial effect in the overdose setting. PMID- 16702901 TI - Quetiapine for hypersexuality and delusional jealousy after stroke. PMID- 16702902 TI - Hypothermia in a patient receiving risperidone and paroxetine. PMID- 16702903 TI - Impact of orally disintegrating olanzapine on use of intramuscular antipsychotics, seclusion, and restraint in an acute inpatient psychiatric setting. PMID- 16702904 TI - Altered expression of myeloperoxidase precursor, myeloid cell nuclear differentiation antigen, Fms-related tyrosine kinase 3 ligand, and antigen CD11A genes in leukocytes of clozapine-treated schizophrenic patients. PMID- 16702905 TI - Catechol-O-methyltransferase and monoamine oxidase-A polymorphisms and treatment response to typical and atypical neuroleptics. PMID- 16702906 TI - Treatment of olfactory hallucinations with topiramate. PMID- 16702907 TI - The effects of topiramate adjunctive treatment added to antidepressants in patients with resistant obsessive-compulsive disorder. PMID- 16702908 TI - Much improved outcome with gabapentin-divalproex combination in adults with bipolar disorders and developmental disabilities. PMID- 16702909 TI - Combination therapy of lamotrigine and escitalopram may cause myoclonus. PMID- 16702910 TI - Levetiracetam for the treatment of alcohol withdrawal syndrome: an open-label pilot trial. PMID- 16702911 TI - Precision and limitation of measuring two-dimensional wear on clinical radiographs. AB - The precision of measuring polyethylene penetration in total hip arthroplasties has been studied using phantom models and retrieved cups, but not on radiographs obtained in vivo. We used Hip Analysis Suite Hip32 computer software to measure two-dimensional penetration in two groups of anteroposterior pelvic radiographs taken in vivo. We determined the measurement error for precision, the effect of position change on precision, and compared direct and indirect measurements. The influences on precision with respect to followup time were measured using a separate group of radiographs. Precision was 0.89 to 1.16 mm and decreased as more variables were added. The direction of wear was toward the contact surface of the cup in greater than 50% of patients with more than 5 years followup. Negative wear severely affected precision in 40% to 50% of patients. Individual penetration measurements were unreliable when total penetration was low (< 1.1 mm). The two-dimensional computer technique has limitations when measuring such low wear. Measurement errors were caused by slight radio-graphic distortions of the femoral head and acetabular component and by blurred edges on some clinical radiographs. PMID- 16702912 TI - Femoral biologic plate fixation. AB - New techniques of fracture fixation such as indirect reduction, careful soft tissue handling, and elastic fixation are being used in place of older methods of mechanical stabilization. These new techniques led to the concept of biologic plate fixation. We systematically reviewed the literature and provide an overall evaluation of femoral biologic fixation for fracture treatment. We analyzed 19 studies with 687 patients with 697 femoral fractures. Twenty-three percent of the fractures were subtrochanteric, 29% were diaphyseal, and 48% were supracondylar. Twenty-two percent were open fractures. The majority (81%) were comminuted and AO Types B and C. The overall union rate was 98.4%, with a mean time to union ranging from 10.7 to 24 weeks. Primary or secondary bone-grafting procedures were reported in all but one study with a frequency ranging from 0-55%. The most frequently recorded complications were malunion (0-29%) and reoperation (0-23%). The high union rate, low infection rate (2%), and occasional need for bone graft indicate biologic plate fixation is a viable alternative to modern nailing techniques, particularly in patients with polytrauma. LEVEL OF EVIDENCE: Therapeutic study, Level III. PMID- 16702913 TI - Case reports: Epiphysiodesis in a patient with osteopetrosis. AB - We describe a 14-year-old girl with intermediate-type osteopetrosis who presented with a 4.8-cm leg-length discrepancy secondary to premature distal femoral growth plate closure. She had a projected leg-length discrepancy of 7 cm at maturity, which was predicted using Moseley charts. The patient was treated successfully with an epiphysiodesis. Her remaining growth was predicted by Moseley charts. A percutaneous epiphysiodesis can treat leg-length discrepancies in patients with osteopetrosis. PMID- 16702914 TI - Knee pain after tibial nailing: the role of nail prominence. AB - We analyzed the relationship between knee pain after tibial nailing and nail prominence. We identified 70 patients in our trauma registry with healed fractures initially treated with intramedullary nails. Subjective pain and function were measured with visual analog pain scales and Lysholm knee scores at a mean of 20 months after fracture. These scores were compared with nail prominence measured on postoperative radiographs. More than 49% of patients had knee pain. Subjective knee pain was more common in women and patients with a smaller plateau width. Anterior nail prominence was associated with increased pain at rest. Patients with superior nail prominence had increased pain with kneeling and walking. Nail prominence correlated with increased knee pain. We think surgeons can decrease, but not eliminate, the severity of knee pain after tibial nailing by burying the tip of the nail as reflected on lateral radiographs. LEVEL OF EVIDENCE: Prognostic Study, Level II-1 (retrospective study). See the Guidelines for Authors for a complete description of levels of evidence. PMID- 16702915 TI - The role of the ossific nucleus in the treatment of established hip dislocation. AB - Timing the reduction of a delayed presenting dislocated hip is controversial if the ossific nucleus of the proximal femoral epiphysis is absent. We formulated a decision model for management of 6- to 13-month-old infants based on two strategies: waiting for the ossific nucleus to appear before reducing the hip or immediate reduction. The model included the occurrence of long-term physical disability within a period of 20 years. A literature synthesis provided outcome probabilities. Outcome was measured by utilities derived by content experts. Waiting for the ossific nucleus was the preferred strategy with an expected value of 0.95 as opposed to 0.86 in the immediate reduction strategy. Sensitivity analyses showed the model was robust. Based on the results of decision analysis, reducing a dislocated hip in the presence of the ossific nucleus is likely to be the better strategy if avascular necrosis and long-term disability are considered. The difference between the two strategies is equivalent to one quality-adjusted life year, which is substantial. LEVEL OF EVIDENCE: Economic and Decision Analyses, Level II-1. See the Guidelines for Authors for a complete description of levels of evidence. PMID- 16702918 TI - Case reports: management of large segmental tibial defects using a cylindrical mesh cage. AB - We report a case series of three patients who sustained open Gustilo-Anderson Type IIIB tibia fractures associated with extensive segmental bone and soft tissue loss. The patients initially were treated with serial wound irrigations, debridements, and external fixation. After the soft tissue envelope was reconstructed successfully, each large segmental bone defect was reconstructed with a cylindrical titanium mesh cage packed with a composite of cancellous allograft and demineralized bone matrix putty and stabilized with a statically locked intramedullary nail. The mean segmental bone loss was 12.2 cm, and all patients had a minimum 1-year followup. One year after reconstruction, radiographs showed stable, well-aligned, healed constructs, and computed tomography images verified the presence of bony ingrowth throughout the cages. All patients were able to ambulate with full weightbearing, and had good ipsilateral knee, hip, and ankle range of motion. This technique seems to be a reasonable alternative for treating large segmental tibial bone defects. PMID- 16702921 TI - Unicompartmental knee arthroplasty in octogenarians: survival longer than the patient. AB - The unicompartmental knee arthroplasty continues to gain popularity as a viable treatment option for disease isolated to one compartment. It has been reported to provide decreased perioperative morbidity, faster recovery, and excellent long- term survival. We hypothesized that the unicompartmental knee arthroplasty is durable enough to benefit octogenarians, and may be a viable alternative to total knee arthroplasty as the definitive treatment of localized arthritis in this age group. From 1978 to 1990, 28 consecutive patients (38 knees) 80 years or older had unicompartmental knee arthroplasties. Knee Society knee and function scores improved at an average of 4 years followup (range, 2-9 years). Family members reported 90% patient satisfaction regarding expectations and desire to have the surgery again. The mean postoperative survival was 11.9 years, and only two of the 38 knees (5%) required surgical intervention. At final followup, 25 patients had died with all but one patient having the index unicompartmental knee arthroplasty in place and functioning well. Of the three living patients, one required surgery for femoral component fracture 10 years after the index procedure. The unicompartmental knee arthroplasty can be expected to provide reliable and durable results in certain octogenarians, and should be regarded as a definitive treatment option in appropriated selected patients of this age group. PMID- 16702922 TI - Bone cement improves suture anchor fixation. AB - Suture anchor fixation failure can occur if the anchor pulls out of bone. We hypothesized that suture anchor fixation can be augmented with polymethylmethacrylate cement, and that polymethylmethacrylate can be used to improve fixation in a stripped anchor hole. Six matched cadaveric proximal humeri were used. On one side, suture anchors were placed and loaded to failure using a ramped cyclic loading protocol. The stripped anchor holes then were injected with approximately 1 cc polymethylmethacrylate, and anchors were replaced and tested again. In the contralateral humerus, polymethylmethacrylate was injected into anchor holes before anchor placement and testing. In unstripped anchors, polymethylmethacrylate increased the number of cycles to failure by 34% and failure load by 71% compared with anchors not augmented with polymethylmethacrylate. Polymethylmethacrylate haugmentation of stripped anchors increased the cycles to failure by 31% and failure load by 111% compared with unstripped uncemented anchors. No difference was found in cycles to failure or failure load between cemented stripped anchors and cemented unstripped anchors. Polymethylmethacrylate can be used to augment fixation, reducing the risk of anchor pull-out failure, regardless whether the suture anchor hole is stripped or unstripped. PMID- 16702924 TI - [The role of podocytes in normal glomerular function and in the pathogenesis of glomerulonephritis. Part I. Phenotypic and functional characteristics of podocytes during their differentiation and maturity]. AB - Podocytes play an important role in glomerular function. Together with endothelial cells of the glomerular capillary loop and the glomerular basement membrane they form a filtration barrier. Podocytes cooperate with mesangial cells to support the structure and function of the glomerulus. With regard to its structure, the podocyte can be divided into three structural and functional segments: cell body, major processes, and foot processes, the latter playing a crucial role in the pathogenesis of proteinuria. Podocytes are end-differentiated cells with specific phenotypic features associated with their function. The compensatory ability of these cells is limited to hypertrophy. Under normal conditions a podocyte cannot restart the cell cycle. Attention has recently been drawn to the importance of slit diaphragms in the pathogenesis of proteinuria. Structural damage to each of the slit diaphragm proteins as well as disturbances in their mutual interactions can result in insufficiency of the filtration barrier. PMID- 16702923 TI - Tracking expression of virally mediated BMP-2 in gene therapy for bone repair. AB - Ex vivo gene therapy using stem cells transduced with viral vectors is a useful method for delivering a therapeutic protein to augment bone repair in animal models. However, the duration of cell-mediated protein production and the fate of the transduced cells are unknown. We constructed an adenoviral vector encoding Myc epitope tagged bone morphogenetic protein (BMP)-2 gene (AdBMP-2). Rat bone marrow cells transduced with this vector produced biologically active BMP-2 protein, which was confirmed by Western blot analysis and alkaline phosphatase assay. Implantation of bone marrow cells infected ex vivo with AdBMP-2 caused orthotopic bone formation in mouse hindlimbs and bony union of critical-sized mouse radial defects. Immunohistochemical analysis revealed that rBMCs expressed Myc epitope-tagged BMP-2 protein for 14 days in vivo and became incorporated in the endochondral fracture callus. This novel adenovirus encoding for epitope tagged BMP-2 can be used for immunohistochemical tracking of transduced cells in ex vivo gene therapy for bone repair. PMID- 16702926 TI - [Platelet-mediated regulation of immunity]. AB - Platelets, essentially non-nucleated blood cells, are highly reactive components of the circulatory system. They have long been consigned solely to a role in thrombosis and hemostasis. Platelets release platelet-derived microvesicles (PMV), also known as platelet-derived microparticles (PMP). They are important but still under-appreciated mediators of intracellular cross-talk between platelets and other cells and modulate their function. They are formed after stimulation of platelets by an agonist, such as thrombin, collagen, and many others. Platelets can carry biological mediators; they can modulate immune responses and inflammatory events. Microparticles can be released from activated platelets and leukocytes in inflammatory events, but in atherosclerosis they can be released from apoptotic endothelial cells, lymphocytes, and monocytes. The markers of innate immunity expressed by platelets are Toll-like receptors (TLRs). The role of platelets in tumor progression and metastasis has been recognized, but the mechanism of their action remains still unclear. PMID- 16702925 TI - [The role of podocytes in normal glomerular function and in the pathogenesis of glomerulonephritis. Part II. Phenotypic and functional changes of podocytes in glomerulonephritis]. AB - Recent results indicate that intrinsic glomerular cells (podocytes, mesangial and endothelial cells) are active participants in the inflammatory process in the glomerulus. Particular attention is drawn to podocyte injury in glomerulonephritis. Podocytes are end-differentiated cells which have lost their proliferation abilities and can only compensate by hypertrophy. The inability to proliferate is the cost which a podocyte must pay for the development of highly specialized structures and ability to adhere to the glomerular basement membrane. Collapsing focal-segmental glomerulosclerosis is a condition in which podocytes proliferate, but this process is associated with loss of their maturity markers as well as with abnormalities in the expressions of cell cycle proteins. Dysfunction of slit diaphragms is an important element in the pathogenesis of glomerulopathies with marked proteinuria. Recent studies also underline the importance of neprilysin in the pathogenesis of glomerulonephritis. This is the first podocytic antigen which has been shown to induce human membranous glomerulonephritis. PMID- 16702927 TI - Endoscopic thoracic sympathectomy for palmar hyperhidrosis. PMID- 16702928 TI - Omission of mediastinal lymph node dissection in lung cancer: its techniques and diagnostic procedures. AB - To reduce or omit a mediastinal lymph node dissection in the patients with clinical stage I non-small cell lung cancer (NSCLC), several authors examined the prevalence of metastatic sites of lymph nodes. Because lymphatic drainage usually heads for the upper mediastinum in upper lobe cancer and for the lower mediastinum in lower lobe cancer, upper and lower mediastinal lymph node dissection could be reduced in lung cancers of lower lobe and upper lobe. By using sentinel node (SN) navigation surgery, it is possible to omit mediastinal lymph node dissection. Radiological findings are also useful to determine reduction of mediastinal lymph node dissection. In clinical stage Ia adenocarcinomas that show ground glass opacity (GGO) findings on computed tomography (CT) or negative for fluorodeoxyglucose accumulation on positron emission tomography (PET), mediastinal lymph node dissection can be omitted, because these types of adenocarcinomas rarely metastasize to the lymph nodes. By using these procedures, mediastinal lymph node dissection can be reduced or omitted with little risk of local recurrence. PMID- 16702929 TI - FDG-PET imaging for lymph node staging and pathologic tumor response after neoadjuvant treatment of non-small cell lung cancer. AB - PURPOSE: A number of studies have demonstrated that 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) is effective for staging of lung cancer. However, the efficacy of FDG-PET for staging lung cancer after neoadjuvant treatment is still controversial. This study compared FDG-PET and computed tomography (CT) for lung cancer staging, and evaluated the ability of the two methods to predict the pathologic response of the primary tumor to neoadjuvant treatment. PATIENTS AND METHODS: Twenty-two patients who underwent neoadjuvant treatment followed by surgery were investigated. Eighteen patients received chemoradiotherapy and four patients received chemotherapy only. One hundred and three lymph node stations in the 22 patients were evaluated by FDG-PET and CT. The pathologic responses of the tumors were compared by FDG-uptake and tumor size on CT for the 15 patients who underwent FDG-PET and CT both before and after neoadjuvant treatment. RESULTS: There was no significant difference in the ability of FDG-PET or CT to predict residual viable tumor. Although positive predictive value by FDG-PET (0.29) was lower than that by CT (0.64) (p=0.04) in the mediastinal lymph nodes, there were no statistically significant differences in the other results of lymph nodes by FDG-PET and CT. Both decrease in FDG uptake and decrease in tumor size by CT after neoadjuvant treatment correlated significantly with pathologic response in the 15 patients (p=0.003 and 0.009, respectively). CONCLUSION: FDG-PET did not appear to offer any advantages over CT for lymph node staging or for predicting the pathologic response after neoadjuvant treatment of non-small cell lung cancer. PMID- 16702931 TI - Measurement of cerebral-oxygenation status when commencing cardiopulmonary bypass in pediatric open-heart surgery. AB - OBJECTIVE: We hypothesize that there is a difference in the cerebral-oxygenation status between cyanotic and non-cyanotic congenital heart disease when commencing a crystalloid-primed cardiopulmonary bypass (CPB). We tested this hypothesis by using near-infrared spectroscopy (NIRS). METHODS: Group 1 consisted of ten patients with non-cyanotic congenital heart diseases, including atrial septal (n=4) and ventricular septal defects (n=6), while group 2 consisted of ten patients with cyanotic congenital heart diseases, including tetralogy of Fallot (n=7) and univentricular heart (n=3). Changes in cerebral-oxygenated, deoxygenated and total hemoglobin concentrations were measured by NIRS just before and every minute for the first 10 min after commencing CPB. Arterial blood analysis was performed at those same time times. RESULTS: NIRS showed a rapid fall and plateauing of cerebral-oxygenated, deoxygenated and total hemoglobin in group 1. However, although group 2 showed a rapid fall and plateauing of cerebral oxygenated hemoglobin, a rapid fall and continuous gradual decrease in cerebral deoxygenated and total hemoglobin were also seen. Cerebral-deoxygenated and total hemoglobin decreased more markedly in group 2 than in group 1 (P<0.001, 0.01, respectively). CONCLUSION: NIRS revealed that the cerebral-oxygenated hemoglobin could be maintained at a similar level at the beginning of CPB in both groups. However, it showed a different distribution of cerebral-deoxygenated and total hemoglobin between the groups. An inadequate cerebral-oxygenation status may occur in the early phase of CPB in patients with cyanotic congenital heart diseases. PMID- 16702930 TI - Cardioprotective effects of tetrahydrobiopterin in cold heart preservation after cardiac arrest. AB - BACKGROUND: It has recently been shown that tetrahydrobiopterin (BH4), an essential cofactor of nitric oxide synthase (NOS), reduces ischemia-reperfusion myocardial injury. The aim of this study was to determine if supplementation with BH4 after cardiac arrest followed by cold heart preservation would exert a cardioprotective effect against ischemia-reperfusion injury. MATERIALS AND METHODS: Isolated perfused rat hearts were subjected to 4 degrees C cold ischemia and reperfusion. Hearts were treated with cold cardioplegic solution with or without BH4 just before ischemia and during the first 5 min of reperfusion period. Effects of BH4 on left ventricular function, myocardial contents of high energy phosphates, and nitrite plus nitrate were measured in the perfusate, before ischemia and after reperfusion. Moreover, the effect of BH4 on the cold heart preservation followed by normothermic (37 degrees C) ischemia was determined. RESULTS: BH4 improved the contractile and metabolic abnormalities in reperfused cold preserved hearts that were subjected to normothermic ischemia. Furthermore, BH4 significantly alleviated ischemic contracture during ischemia, and restored the diminished perfusate levels of nitrite plus nitrate after reperfusion. CONCLUSION: These results demonstrated that BH4 reduces ischemia reperfusion injury in cold heart preservation. The cardioprotective effect of BH4 implies that BH4 could be a novel and effective therapeutic option in the preservation treatment of donor heart after cardiac arrest. PMID- 16702932 TI - A case complicated with Horner's syndrome after off-pump coronary artery bypass. AB - In the present report, a case complicated with Horner's syndrome after off-pump coronary artery bypass (OPCAB) was presented. This case showed ptosis and miosis in the left eye promptly after OPCAB. No abnormal neurological findings other than Horner's syndrome were observed in postoperative examinations including head magnetic resonance imaging (MRI), and this case was thought to have Horner's syndrome as a complication after cardiac surgery through median sternotomy. PMID- 16702933 TI - Postoperative chylothorax in patients with a thoracic aortic aneurysm. AB - OBJECTIVES: Postoperative chylothorax in patients with a thoracic aneurysm is generally infrequent. We report a mode of surgery to reduce the incidence of paraplegia. We review our experience with chylothorax after resection of an aneurysm to find its cause and to evaluate the success of management. PATIENTS AND METHODS: For descending thoracic aneurysms, intercostal arteries in the aneurysm were exposed before incising the aneurysm and, they were only sacrificed no change in motor-evoked potentials (MEPs) occurred after temporary occlusion. Between January 2001 and December 2003, out of a total of 147 aneurysms including thoracic and thoracoabdominal, 4 consecutive patients (2.7%) with chylothorax were reviewed. RESULTS: The chylothorax was diagnosed 1.5 days after operation (range 1 to 2 days). All patients were initially treated by the cessation of oral intake. This treatment was successful for 2 patients and the remaining 2 required surgical intervention to control the fistula. Chylothorax was cured in all patients. CONCLUSION: We postulate that chylothorax is caused by injury of the thoracic duct or its branch during the exposing the intercostal arteries. In the management of chylothorax, early intervention is recommended when the volume of chylous fluid is not decreased by conservative treatment. PMID- 16702934 TI - Morphological study of abdominal aortic aneurysm: optimal stent-graft size for Japanese patients. AB - PURPOSE: We report on the optimal stent-graft (SG) size for Japanese patients with abdominal aortic aneurysm (AAA). MATERIALS AND METHODS: Ninety three Japanese patients undergoing elective AAA repair were selected for this study. The parameters measured were the proximal neck (PN) diameter (D1), the diameter of the right and left common iliac arteries (D2 and D3, respectively), the diameter of the right and left external iliac arteries (D4 and D5, respectively), the distance between the lowest renal artery and the common iliac arterial bifurcation (L1), and the distance between the right and left common iliac arterial bifurcations and the internal iliac arterial bifurcation (L2 and L3, respectively). RESULTS: The following results were obtained: D1: 20.7+/-3.9 mm (14 to 28 mm); D2: 14.0+/-3.0 mm (9.5 to 20 mm); D3: 13.8+/-3.1 mm (9 to 19.5 mm); D4: 7.5+/-1.0 mm (6 to 10 mm); D5: 7.4+/-0.9 mm (6 to 10 mm); L1: 107.7+/ 13.4 mm (80 to 130 mm); L2: 40.0+/-10.1 mm (20 to 61 mm); L3: 39.7+/-9.6 mm (20 to 60 mm). CONCLUSION: The results indicate the necessity of exercising adequate care when selecting a device for Japanese patients. PMID- 16702935 TI - Primary osteosarcoma of the lung: a case report. AB - We report a rare case of primary osteosarcoma of the lung. A 73-year-old Japanese man with a productive cough and hemosputum was referred to us for further evaluation of a huge cavitating mass in the left upper lobe, shown on a radiograph of his chest. The result of a tumor biopsy, via fiberoptic bronchoscope, raised a strong suspicion of sarcoma. Therefore a left upper lobectomy was performed without any adjuvant therapy. The tumor, which measured 72 x 70 x 62 mm, was well-defined, whitish-yellow in color and soft in consistency. Histological examination of the tumor showed a dense proliferation of spindle cells and the presence of many collagen fibers. Eosinophilic osteoid, with no epithelial structures, were noted in the stroma. Immunohistochemically, the tumor cells were positive for mesenchymal, but negative for epithelial markers. These pathological features suggested the tumor was an osteosarcoma. A general inspection of other organs did not reveal any more tumorous lesions, therefore, the final diagnosis of the tumor was primary osteosarcoma of the lung. PMID- 16702936 TI - Kissing pleural metastases from metastatic osteosarcoma of the lung. AB - Two patients with osteosarcoma lung metastases of which migrated to the parietal pleura due to contact are reported. The first patient was a 16-year-old male who had a pleural metastasis in the diaphragm within an area in contact with a single lung metastasis. Both of the tumors were resected, followed by systemic chemotherapy. Nine months after the resection of the first metastases, two other lung metastases were found which were resected after chemotherapy. The patient is alive without recurrence 84 months after the first resection of the metastases. The second patient was an 11-year-old female with a pleural metastasis of osteosarcoma which was within an area in contact with a single lung metastasis, which had been resected 4 months before. We concluded (1) that a lung metastasis of osteosarcoma occasionally metastasizes to the pleura due to contact; and (2) that because this kissing metastases of osteosarcoma could be cured by a complete resection, the intrathoracic cavity should be thoroughly observed. PMID- 16702937 TI - Extrathoracic neurilemoma of the lateral chest wall mimicking a subcutaneous tumor: report of a case. AB - Neurilemomas of the chest wall are usually solitary lesions which bulge toward the pleural cavity in the posterior mediastinum. We report a rare case of neurilemoma growing outside the thorax from the intercostal nerve. A 33-year-old man was admitted with a chest wall bulging mass. His past history was unremarkable. Computed tomography showed a well-circumscribed mass toward the extrathorax adjacent to the ninth intercostal space. Surgical resection was performed for a firm diagnosis and treatment. This tumor was easily dissected from the surrounding tissues without passing into the pleural cavity by percutaneous approach, and then completely resected. He has been followed-up for 16 months without recurrence. PMID- 16702938 TI - Long-term survival of Askin tumor for 10 years with 2 relapses. AB - An abnormal shadow was noted on a chest X-ray of a 32-year-old female in a medical check-up in March 1995, 3 months after she had given birth. Thoracic CT detected a tumor in contact with the left thoracic wall, and tumorectomy was performed in May 1995. The tumor was diagnosed as a primitive neuroectodermal tumor (PNET). After surgery, the thoracic wall to which the tumor adhered was treated with irradiation at 50 Gy. Chemotherapy was considered, but the patient did not wish to undergo this treatment. Lung metastasis occurred 5 years after the first surgery, and the left lower lobe of the lung was partially resected. Four years later, lobectomy of the left lower lobe of the lung was performed, due to further lung metastasis. The patient remains healthy as of April 2005. PMID- 16702939 TI - Completely wide resection of malignant fibrous histiocytoma of the chest wall; expect for long survival. AB - Malignant fibrous histiocytoma (MFH) rarely occurs in the chest wall. A case of MFH originating from the chest wall is herein reported. We performed radical en block resection of the whole chest wall together with the tumor and reconstructed it with Marlex mesh. There was no recurrence 4 years after operation. We consider radical en-block resection for MFH and reconstruction with Marlex mesh a safe operation and may provide a long-term survival. PMID- 16702940 TI - Cardiac angiosarcoma with right atrial perforation and cardiac tamponade. AB - Primary cardiac angiosarcoma is a rare tumor associated with a poor prognosis. We report a case of a 59-year-old woman with right atrial angiosarcoma presenting with cardiac tamponade due to right atrial perforation. She underwent urgent surgical resection of the tumor. However, the patient died 68 days after surgery due to local recurrence. An effective treatment for cardiac angiosarcoma has not yet been established. However, more aggressive treatment with a combination of surgery, radiation, chemotherapy and IL-2 should be considered. PMID- 16702941 TI - Isthmus of a horseshoe kidney overlying a ruptured abdominal aortic aneurysm: a case report. AB - Horseshoe kidney is a rare congenital anomaly showing various degrees of fusion and accessory blood supply. Coexistence of horseshoe kidney and aortic aneurysm therefore presents a technical challenge to vascular surgeons. We report an 83 year-old woman with a ruptured abdominal aortic aneurysm associated with horseshoe kidney. Preoperative computed tomography (CT) showed discontinuity of the aneurysm wall in relation to the isthmus of the horseshoe kidney, and rupture of the aneurysm into the retroperitoneal space. The aneurysm was exposed via a transperitoneal approach, and grafting was performed successfully together with division of the isthmus of the horseshoe kidney. Renal function showed no impairment postoperatively. PMID- 16702942 TI - Abdominal aortic grafting for renal and lower extremity hypoperfusion due to aortic dissection with progressive thrombosis of a false channel. AB - We present the case of a 53-year-old man with renal and bilateral limb ischemia due to Stanford B aortic dissection. The thrombosis of the false lumen had progressed and compressed the true lumen, developing renal and leg ischemia. Urgent graft replacement of the infrarenal abdominal aorta with proximal fenestration was successfully performed and the patient was discharged without complications. PMID- 16702943 TI - Pediatric Berlin heart excor. PMID- 16702944 TI - Antileukemic roles of human phospholipid scramblase 1 gene, evidence from inducible PLSCR1-expressing leukemic cells. AB - Phospholipid scramblase 1 (PLSCR1) is a multiply palmitoylated protein which is localized in either the cell membrane or nucleus depending on its palmitoylated state. The increasing evidence showed the biological roles of PLSCR1 in cell signaling, maturation and apoptosis. To investigate the functions of PLSCR1 in leukemic cells, we generated an inducible PLSCR1-expressing cell line using myeloid leukemic U937 cells. In this cell line, PLSCR1 was tightly regulated and induced upon tetracycline withdrawal. Our results showed that inducible PLSCR1 expression arrested the proliferation of U937 cells at G1 phase. Meanwhile, PLSCR1-overexpressing U937 cells also underwent granulocyte-like differentiation with increased sensitivity to etoposide-induced apoptosis. Furthermore, we also found that PLSCR1 induction increased cyclin-dependent kinase inhibitors p27(Kip1) and p21(Cip1) proteins, together with downregulation of S phase kinase associated protein 2 (SKP2), an F-box subunit of the ubiquitin-ligase complex that targets proteins for degradation. Additionally, PLSCR1 induction significantly decreased c-Myc protein and antiapoptotic Bcl-2 protein. Although the exact mechanism by which PLSCR1 regulates these cellular events and gene expression remains unresolved, our results suggest that PLSCR1 plays the antagonistic role regarding leukemia development. These data will shed new insights into understanding the biochemical and biological functions of PLSCR1 protein. PMID- 16702945 TI - Induction of melanoma cell apoptosis and inhibition of tumor growth using a cell permeable Survivin antagonist. AB - The inhibitor of apoptosis gene family member Survivin is highly expressed in most tumors, and appears to be a promising target for cancer therapy. Although a variety of Survivin antagonists have been shown to induce apoptosis in malignant cells, the potential utility of these agents is limited by inefficient delivery and cell impermeability. We generated recombinant fusion proteins containing the TAT protein transduction domain and either wild-type Survivin (TAT-Surv-WT) or a dominant-negative mutant (TAT-Surv-T34A). The TAT-Surv proteins were purified by sequential affinity and ion-exchange chromatography, and at 30 nM concentration demonstrated rapid entry into cells at 30 min. Whereas TAT-Surv-WT had minimal effect on YUSAC2 or WM793 melanoma cells, TAT-Surv-T34A induced cell detachment, DNA fragmentation, caspase-3 activation and mitochondrial release of apoptosis inducing factor at low microM concentrations. Intraperitoneal (i.p.) injection of mice bearing subcutaneous YUSAC2 xenografts with TAT-Surv-T34A (10 mg/kg) was associated with rapid tumor accumulation at 1 h, and increased tumor cell apoptosis and aberrant nuclei formation in situ. Repeated i.p. injection of TAT Surv-T34A resulted in a 40-50% reduction in growth and mass of established tumors, compared to those similarly injected with saline buffer or TAT-Surv-WT. These studies demonstrate the feasibility of systemic tumor treatment using a cell-permeable Survivin antagonist. PMID- 16702946 TI - Isolation and characterization of dominant and recessive IL-3-independent hematopoietic transformants. AB - Retroviral integration mutagenesis and treatment with the frameshift mutagen ICR191 were used to transform v-H-ras expressing PB-3c cells to interleukin-3 (IL 3) independence. Six clones displayed viral integrations into the 3' region of the IL-3 gene thus acting post-transcriptionally by disrupting the AU-rich instability element. Two clones contained reverse orientation integration into the raf-1 gene revealing an enhancer insertion mechanism. Growth by this mechanism was sensitive to the Raf-1 inhibitor BAY 43-9006 and the Mek inhibitor U0126. Following treatment with ICR191, IL-3-independent clones were recovered and studied by cell fusion. With 21/22 clones, IL-3 independence resulted from a recessive mechanism as cellular hybrids with parental cells reverted to IL-3 dependence. Recessive clone D2c displayed increased phospho-Erk1/2 levels and was growth sensitive to U0126, but not to BAY43-9006. The single dominant clone, D5a, showed no signs of mitogen-activated protein kinases pathway activation but displayed constitutive phosphorylation of Stat5. We conclude that PB-3c has several options to acquire IL-3 growth autonomy involving transcriptional or post transcriptional mechanisms affecting the distal regulators Erk or Stat5. The reported panel of independent dominant and recessive transformants should provide a useful tool for inhibitor profiling. PMID- 16702947 TI - c-Abl phosphorylates Hdm2 at tyrosine 276 in response to DNA damage and regulates interaction with ARF. AB - The p53 tumour-suppressor protein is tightly regulated through its association with the Hdm2 E3 ligase. Activation of p53 by DNA strand breaks is orchestrated by the ataxia-telangiectasia mutated (ATM) protein kinase and involves interruption of Hdm2-mediated p53 degradation. As part of this mechanism ATM itself, and the ATM-activated protein tyrosine kinase, c-Abl, inhibit Hdm2 function through phosphorylation of serine 395 and tyrosine 394 (Y394), respectively. In the present study, we have identified a novel target of c-Abl in the Hdm2 protein, tyrosine 276 (Y276). We show that c-Abl phosphorylates this residue in vitro and confirm that Y394 is a target of c-Abl. We also show that Y276 is phosphorylated in a c-Abl-dependent manner in cultured cells and provide evidence that Y276 is phosphorylated in response to DNA damage coincident with the activation of c-Abl. Finally, we show that Y276 phosphorylation stimulates interaction with ARF, leading to increased levels of nucleolar Hdm2 and decreased turnover of p53. These data establish Y276 as a physiological target of c-Abl that contributes functionally to the induction of p53. PMID- 16702948 TI - A selective inhibitor of the p110delta isoform of PI 3-kinase inhibits AML cell proliferation and survival and increases the cytotoxic effects of VP16. AB - Current therapy for acute myeloid leukaemia (AML) is suboptimal with a high incidence of relapse. There is strong evidence that constitutive phosphoinositide 3-kinase (PI3K) activity plays a significant role in the pathophysiology of AML. PI3K products are derived from the activity of a number of PI3K catalytic isoforms (class I, II and III) but the relative contribution of these enzymes in AML remains unknown. As non-isoform-selective inhibitors of PI3K such as LY294002 may produce unwanted toxicity to normal tissues, we have investigated the role of the leukocyte-restricted p110delta PI3K isoform in 14 cases of AML. p110delta was detected in all cases whereas the expression levels of the other class I PI3Ks varied more widely, and were often undetectable. The p110delta-selective compound IC87114 inhibited constitutive phosphorylation of the PI3K target Akt/PKB and reduced cell number to a mean of 66+/-5% (range 14-88%). In eight cases, the combination of IC87114 and VP16 (a topoisomerase II inhibitor) was synergistic in reducing viable cell number, and was associated with a reduction in constitutive NF-kappaB activity. IC87114 did not have direct adverse effects or enhance the activity of VP16 on the proliferation and survival of normal haemopoietic progenitors. Overall, our results identify the p110delta isoform as a potential therapeutic target in AML and support a clinical approach to use isoform selective over broad-spectrum PI3K inhibitors. PMID- 16702949 TI - Enhanced metastatic dissemination to multiple organs by melanoma and lymphoma cells in timp-3-/- mice. AB - Identifying versatile inhibitors of metastasis that operate in multiple sites against distinct cancer cell types is important for designing novel therapeutics for metastasis. We show that multiple tissues of timp-3-/- mice are more susceptible to metastatic colonization. Overall, a 5-14-fold increase in liver and kidney colonization occurred by EL-4 lymphoma cells, and a twofold increase upon targeting B16F10 melanoma cells to the bone or lung of timp-3-/- mice. There was a general lack of macrophage or neutrophil localization to metastases in the liver, kidney and lung, and of osteoclasts to bone in both genotypes. Analysis of lung showed that proliferation or angiogenesis were unaltered within the metastatic colonies. Lung-trap assays revealed that initial tumor cell trapping was similar in the lung vasculature of timp-3-/- and wild-type mice. However, more tumor cells were found in timp-3-/- lungs at 48 and 96 h after tumor cell injection indicating more efficient extravasation and initial proliferation. Activation of pro-MMP-2 was greater in timp-3-/- lungs at these time points. These data demonstrate TIMP-3 functions to inhibit metastatic dissemination of diverse cancer cells to multiple organs. TIMP-3 regulates MMP-2 activation to limit tumor cell extravasation and subsequent colonization of the lung, without augmenting inflammatory cell response. PMID- 16702950 TI - EGFRvIII undergoes activation-dependent downregulation mediated by the Cbl proteins. AB - The overexpression or mutation of tyrosine kinases (TKs), such as the epidermal growth factor receptor (EGFR), can lead to the development of cancer. The most common mutation of the EGFR in glioblastomas is the deletion of exons 2-7 known as the EGFRvIII. This mutant receptor cannot bind EGF but, instead, is constitutively active. The Cbl family of ubiquitin ligases (Cbl, Cbl-b, and Cbl c) targets the activated EGFR for degradation. As the EGFRvIII is transforming, we investigated whether it could be downregulated by the Cbl proteins. The overexpression of all three Cbl proteins resulted in the ubiquitination and degradation of the EGFRvIII. As with the wild-type EGFR, the TK-binding domain and the RING finger of Cbl-b are sufficient for the downregulation of the EGFRvIII. Also, we found that Cbl-b is recruited to the EGFRvIII and inhibits the transformation of NIH 3T3 cells by the EGFRvIII. Mutation of the Cbl-binding site (Y1045F) in the EGFRvIII inhibits its ubiquitination and downregulation by Cbl-b and enhances its ability to transform. Furthermore, the EGFR TK inhibitor, AG 1478, prevents the downregulation of the EGFRvIII by the Cbl proteins and antagonizes the ability of an immunotoxin directed against the EGFRvIII to kill cells expressing this receptor. In conclusion, the EGFRvIII does not transform by escaping regulation by Cbl proteins and this activation-induced downregulation of the EGFRvIII has an important role in mediating the toxicity of anti-EGFRvIII immunotoxins. PMID- 16702951 TI - Tuberin activates the proapoptotic molecule BAD. AB - TSC1, encoding hamartin, and TSC2, encoding tuberin, are tumor suppressor genes responsible for the autosomal dominantly inherited disease tuberous sclerosis (TSC). TSC affects approximately 1 in 6000 individuals and is characterized by the development of tumors, named hamartomas, in different organs. Hamartin and tuberin form a complex, of which tuberin is assumed to be the functional component. The TSC proteins have been implicated in the control of cell cycle and cell size. In addition to enhanced growth, reduced death rates can lead to tumor development. Therefore, defects in the apoptosis-inducing pathways contribute to neoplastic cell expansion. Here, we show that tuberin triggers apoptosis, accompanied by downregulation of p70S6K activity and of phosphorylation of BAD on residue Ser136, and by upregulation of the interaction of BAD/BCL-2 and BAD/BCL XL. AKT phosphorylation negatively regulates tuberin's potential to trigger apoptosis. Experiments with BAD-/- cells demonstrate BAD to be a mediator of tuberin's effects on the regulation of apoptosis. Tuberin interferes with insulin like growth factor-1-induced BAD Ser136 phosphorylation and cell survival. Our work proposes a model in which tuberin-mediated inhibition of p70S6K activates BAD to heterodimerize with BCL-2 and BCL-XL to promote apoptosis. A mutation of TSC2--as it occurs in TSC patients--attenuates this proapoptotic potential, underscoring the relevance of our findings for human pathophysiology. PMID- 16702953 TI - Unique role of SNT-2/FRS2beta/FRS3 docking/adaptor protein for negative regulation in EGF receptor tyrosine kinase signaling pathways. AB - The membrane-linked docking protein SNT-2/FRS2beta/FRS3 becomes tyrosine phosphorylated in response to fibroblast growth factors (FGFs) and neurotrophins and serves as a platform for recruitment of multiple signaling proteins, including Grb2 and Shp2, to FGF receptors or neurotrophin receptors. We previously reported that SNT-2 is not tyrosine phosphorylated significantly in response to epidermal growth factor (EGF) but that it inhibits ERK activation via EGF stimulation by forming a complex with ERK2. In the present report, we show that expression of SNT-2 suppressed EGF-induced cell transformation and proliferation, and expression level of SNT-2 is downregulated in cancer. The activities of the major signaling molecules in EGF receptor (EGFR) signal transduction pathways, including autophosphorylation of EGFR, were attenuated in cells expressing SNT-2 but not in cells expressing SNT-2 mutants lacking the ERK2 binding domain. Furthermore, SNT-2 constitutively bound to EGFR through the phosphotyrosine binding (PTB) domain both with and without EGF stimulation. Treatment of cells with MEK inhibitor U0126 partially restored the phosphorylation levels of MEK and EGFR in cells expressing SNT-2. On the basis of these findings, we propose a novel mechanism of negative control of EGFR tyrosine kinase activity with SNT-2 by recruiting ERK2, which is the site of negative feedback loop from ERK, ultimately leading to inhibition of EGF-induced cell transformation and proliferation. PMID- 16702952 TI - A comprehensive study of chromosome 16q in invasive ductal and lobular breast carcinoma using array CGH. AB - We analysed chromosome 16q in 106 breast cancers using tiling-path array comparative genomic hybridization (aCGH). About 80% of ductal cancers (IDCs) and all lobular cancers (ILCs) lost at least part of 16q. Grade I (GI) IDCs and ILCs often lost the whole chromosome arm. Grade II (GII) and grade III (GIII) IDCs showed less frequent whole-arm loss, but often had complex changes, typically small regions of gain together with larger regions of loss. The boundaries of gains/losses tended to cluster, common sites being 54.5-55.5 Mb and 57.4-58.8 Mb. Overall, the peak frequency of loss (83% cancers) occurred at 61.9-62.9 Mb. We also found several 'minimal' regions of loss/gain. However, no mutations in candidate genes (TRADD, CDH5, CDH8 and CDH11) were detected. Cluster analysis based on copy number changes identified a large group of cancers that had lost most of 16q, and two smaller groups (one with few changes, one with a tendency to show copy number gain). Although all morphological types occurred in each cluster group, IDCs (especially GII/GIII) were relatively overrepresented in the smaller groups. Cluster groups were not independently associated with survival. Use of tiling-path aCGH prompted re-evaluation of the hypothetical pathways of breast carcinogenesis. ILCs have the simplest changes on 16q and probably diverge from the IDC lineage close to the stage of 16q loss. Higher-grade IDCs probably develop from low-grade lesions in most cases, but there remains evidence that some GII/GIII IDCs arise without a GI precursor. PMID- 16702954 TI - Differential effects of NF-kappaB on apoptosis induced by DNA-damaging agents: the type of DNA damage determines the final outcome. AB - The transcription factor nuclear factor kappa-B (NF-kappaB) is generally regarded as an antiapoptotic factor. Accordingly, NF-kappaB activation inhibits death ligand-induced apoptosis. In contrast, ultraviolet light B (UVB)-induced apoptosis is not inhibited but even enhanced upon NF-kappaB activation by interleukin-1 (IL-1). This study was performed to identify the molecular mechanisms underlying this switch of NF-kappaB. Enhancement of UVB-induced apoptosis was always associated with increased release of tumour necrosis factor alpha (TNF-alpha), which was dependent on NF-kappaB activation. The same was observed when UVA and cisplatin were used, which like UVB induce base modifications. In contrast, apoptosis caused by DNA strand breaks was not enhanced by IL-1, indicating that the type of DNA damage is critical for switching the effect of NF-kappaB on apoptosis. Surprisingly, activated NF-kappaB induced TNF-alpha mRNA expression in the presence of all DNA damage-inducing agents. However, in the presence of DNA strand breaks, there was no release of the TNF-alpha protein, which is so crucial for enhancing apoptosis. Together, this indicates that induction of DNA damage may have a significant impact on biological effects but it is the type of DNA damage that determines the final outcome. This may have implications for the role of NF-kappaB in carcinogenesis and for the application of NF-kappaB inhibitors in anticancer therapy. PMID- 16702955 TI - Interference with nuclear factor kappa B and c-Jun NH2-terminal kinase signaling by TRAF6C small interfering RNA inhibits myeloma cell proliferation and enhances apoptosis. AB - The tumor necrosis factor receptor (TNFR)-associated factor (TRAF) family of six adaptor proteins (TRAF1-6) links the TNFR superfamily to the nuclear factor kappa B (NF-kappaB) and activator protein-1 (AP-1) transcriptional activators. Unlike other TRAFs, TRAF6 is also involved in Toll-like/interleukin (IL)-1 receptor (TIR) signal transduction. Thus, inhibition of TRAF6 function could interrupt both CD40 (TNFR family) and IL-1 growth signals, pathways critical to myeloma proliferation. To block TRAF6-mediated IL-1 signaling, we constructed small interfering RNA (siRNA) against TRAF6. We found that siRNA targeting the TRAF6 C terminal (siTRAF6C) receptor interaction domain specifically reduced only TRAF6 protein expression, without affecting TRAF2 or 5 levels, and substantially interfered with IL-1-induced NF-kappaB and c-Jun/AP-1 activation. Inhibition by siTRAF6C was concentration-dependent. SiTRAF6C also significantly reduced myeloma proliferation and enhanced apoptosis in a similar dose-dependent fashion in vitro. More importantly, marked siTRAF6C growth inhibition was detected in vivo when these cells were implanted into the bone marrow of irradiated normal mice. In contrast, introduction of siRNA derived from the TRAF6 Zn-finger domain or an irrelevant siRNA construct failed to alter cell growth or cell death. These studies suggest that TRAF6 may be a new molecular target to block cell signal transduction important for the survival and proliferation of multiple myeloma cells. PMID- 16702956 TI - 7-Bromoindirubin-3'-oxime induces caspase-independent cell death. AB - Indirubin, an isomer of indigo, is a reported inhibitor of cyclin-dependent kinases (CDKs) and glycogen synthase kinase-3 (GSK-3) as well as an agonist of the aryl hydrocarbon receptor (AhR). Indirubin is the active ingredient of a traditional Chinese medicinal recipe used against chronic myelocytic leukemia. Numerous indirubin analogs have been synthesized to optimize this promising kinase inhibitor scaffold. We report here on the cellular effects of 7 bromoindirubin-3'-oxime (7BIO). In contrast to its 5-bromo- and 6-bromo- isomers, and to indirubin-3'-oxime, 7BIO has only a marginal inhibitory activity towards CDKs and GSK-3. Unexpectedly, 7BIO triggers a rapid cell death process distinct from apoptosis. 7-Bromoindirubin-3'-oxime induces the appearance of large pycnotic nuclei, without classical features of apoptosis such as chromatin condensation and nuclear fragmentation. 7-Bromoindirubin-3'-oxime-induced cell death is not accompanied by cytochrome c release neither by any measurable effector caspase activation. Furthermore, the death process is not altered either by the presence of Q-VD-OPh, a broad-spectrum caspase inhibitor, or the overexpression of Bcl-2 and Bcl-XL proteins. Neither AhR nor p53 is required during 7BIO-induced cell death. Thus, in contrast to previously described indirubins, 7BIO triggers the activation of non-apoptotic cell death, possibly through necroptosis or autophagy. Although their molecular targets remain to be identified, 7-substituted indirubins may constitute a new class of potential antitumor compounds that would retain their activity in cells refractory to apoptosis. PMID- 16702957 TI - Tristetraprolin regulates Cyclin D1 and c-Myc mRNA stability in response to rapamycin in an Akt-dependent manner via p38 MAPK signaling. AB - The differential expression of the critical cell cycle control proteins cyclin D1 and c-myc has been shown to result in Akt-dependent hypersensitivity of tumor cells to mTOR inhibitors. We have previously demonstrated that the differential utilization of internal ribosome entry sites within the mRNAs of these transcripts allows maintenance of protein synthesis in the face of rapamycin (rapa) exposure in an Akt-dependent manner. Here, we demonstrate that in addition to this mechanism, cyclin D1 and c-myc mRNA stability is also coordinately regulated following rapa treatment depending on Akt activity status. We identify A/U-rich response elements within the 3' untranslated regions (UTRs) of these transcripts, which confer the observed differential stabilities and show that the RNA-binding protein, tristetraprolin (TTP), interacts with these elements. We also present evidence that TTP accumulates in response to rapa exposure, binds to the cis-acting elements within the cyclin D1 and c-myc 3' UTRs and is differentially serine phosphorylated in an Akt-dependent manner. Furthermore, the differential phosphorylation status of TTP results in its sequestration by 14-3-3 proteins in quiescent Akt-containing cells. Finally, siRNA-mediated knockdown of TTP expression or inhibiting a known regulator of TTP phosphorylation, p38 MAP kinase, abolishes the effects on cyclin D1 and c-myc mRNA stability. We assume that the differential control of cyclin D1 and c-myc mRNA stability and translational efficiency constitutes a coordinate response to rapa contributing to the maintenance of expression of these determinants in rapa-resistant quiescent Akt-containing cells following exposure. PMID- 16702959 TI - E-cadherin is a novel transcriptional target of the KLF6 tumor suppressor. AB - The tumor suppressor KLF6 is a member of the Kruppel-like family of transcription factors, which has been implicated in the pathogenesis of several human carcinomas. Uncovering the transcriptional targets relevant for its tumorigenic properties, including cellular proliferation and invasion, will be essential to understanding possible mechanisms by which KLF6 and its antagonistic splice form, KLF6-SV1, regulate this development. To begin defining possible metastatic related pathways, we analysed the effect of KLF6 dysregulation on a recognized suppressor of cellular invasion, E-cadherin. Targeted KLF6 reduction in an ovarian cancer cell line, SKOV-3, resulted in a 50% reduction of E-cadherin expression (P<0.01) and conversely, KLF6-SV1 silencing upregulated E-cadherin approximately fivefold (P<0.0001). These changes resulted from KLF6 directly transactivating the E-cadherin promoter as demonstrated by luciferase promoter assay and chromatin immunoprecipitation (ChIP). KLF6-mediated changes in E cadherin levels were accompanied by downstream changes in both the subcellular localization of beta-catenin and c-myc expression levels. Moreover, and consistent with these experimental findings, patient-derived epithelial ovarian tumors with low KLF6 and high KLF6-SV1 expression ratios had significantly decreased E-cadherin expression (P<0.0001). These combined findings highlight the E-cadherin pathway as a novel and functionally important mediator by which changes in KLF6 and KLF6-SV1 can directly alter ovarian tumor invasion and metastasis. PMID- 16702958 TI - Functional analysis of the regulatory requirements of B-Raf and the B-Raf(V600E) oncoprotein. AB - The BRAF(V600E) mutation is found in approximately 6% of human cancers and mimics the phosphorylation of the kinase domain activation segment. In wild-type B-Raf (B-Raf(wt)), activation segment phosphorylation is thought to cooperate with negative charges within the N-region for full activation. In contrast to Raf-1, the N-region of B-Raf is constitutively negatively charged owing to the presence of residues D447/D448 and the phosphorylation of S446. Therefore, it has been suggested that this hallmark predisposes B-Raf for oncogenic activation. In this study, we demonstrate that neutralizing mutations of these residues (in particular S446 and S447), or uncoupling of B-Raf from Ras-guanine 5' triphosphate (GTP), strongly reduce the biological activity of B-Raf in a PC12 cell differentiation assay. We also confirm that S365 is a 14-3-3 binding site, and determine that mutation of this residue rescues the impaired biological activity of B-Raf proteins with a neutralized N-region, suggesting that the N region opposes a 14-3-3-mediated transition into an inactive conformation. However, in the case of B-Raf(V600E), although complete N-region neutralization resulted in a 2.5-fold reduction in kinase activity in vitro, this oncoprotein strongly induced PC12 differentiation or transformation and epithelial mesenchymal transition of MCF-10A cells regardless of its N-region charge. Furthermore, the biological activity of B-Raf(V600E) was independent of its ability to bind Ras-GTP. Our analysis identifies important regulatory differences between B-Raf(wt) and B-Raf(V600E) and suggests that B-Raf(V600E) cannot be inhibited by strategies aimed at blocking S446 phosphorylation or Ras activation. PMID- 16702960 TI - Errors in the lab and in publications. PMID- 16702964 TI - Breaking the (un)sound barrier: filaggrin is a major gene for atopic dermatitis. AB - We have recently shown that loss-of-function mutations in the filaggrin gene, carried by about 10% of people of European ethnicity, cause ichthyosis vulgaris and are strong predisposing factors for atopic dermatitis and asthma secondary to atopic dermatitis. These results demonstrate a prominent role for the epidermal barrier in atopic disease and have important implications for the study of complex traits. PMID- 16702965 TI - Epidermal differentiation complex yields a secret: mutations in the cornification protein filaggrin underlie ichthyosis vulgaris. AB - Ichthyosis vulgaris (IV), characterized by mild scaling on limbs and lower abdomen, has an incidence of 1 in 250. Smith, McLean, and colleagues demonstrate that common mutations in filaggrin underlie IV. Filaggrin aggregates keratin intermediate filaments and is cross-linked into the cornified envelope to form the epidermal barrier. These findings reinforce the importance of the epidermal barrier in pathogenesis of skin diseases. PMID- 16702966 TI - PSORS1: linking genetics and immunology. AB - Previous studies have localized the psoriasis susceptibility 1 (PSORS1) locus to the proximal major histocompatibility complex class I region, but approximately a dozen genes in the candidate region have thus far been genetically inseparable. According to a team of researchers in the United States and Germany, the primary genetic association has now been worked out, implicating HLA-Cw6 "after all." The results integrate genetics and immunology, further reinforcing our emerging understanding of psoriasis. PMID- 16702967 TI - Cutaneous microdialysis: is it worth the sweat? AB - Microdialysis is a minimally invasive technique for chronological study of metabolic, biochemical, and pharmacological events in living tissue. In the skin, probes are placed in the dermis or subcutis for research in two main areas, percutaneous penetration and various aspects of inflammation. Advances in technique, and the concept of data generation and analysis are leading to new areas of application. PMID- 16702968 TI - The skin as a site of initiation of systemic autoimmune disease: new opportunities for treatment. AB - Dendritic cells are the coordinators of the adaptive immune response. Chronic activation of skin dendritic cells by keratinocyte expression of CD40 ligand (CD40L; CD154) leads to autoimmunity. In this issue, systemic administration of tacrolimus is shown by Loser et al. to effectively treat autoimmunity in a murine model involving transgenic keratinocyte expression of CD40L. PMID- 16702969 TI - "Bak (and Bax) to the future"--of primary melanoma prognosis? AB - Bcl-2 proteins either block or activate the "intrinsic" mitochondrial apoptosis pathway. Loss of expression of proapoptotic Bcl-2 proteins, namely Bax and Bak, in primary melanomas is associated with a worse long-term prognosis. Consequently, inactivation of mitochondrial signaling pathways of apoptosis may not only be a prerequisite for melanoma progression but may also hamper therapeutic efforts with chemotherapeutic drugs. PMID- 16702970 TI - Genomic instability and tumor stem cells. AB - Wang et al. point to the existance of a common progenitor tumor stem cell that gives rise to genomically unstable progeny in malignant melanoma. Although it is not known what creates this genomic instability, given the presence of testis antigens in melanoma, it is tempting to speculate that it is caused by a collision of meiotic and mitotic pathways. PMID- 16702971 TI - More than one gene involved in monilethrix: intracellular but also extracellular players. AB - Monilethrix, an autosomal dominant human hair disorder, is caused by mutations in three type II hair cortex keratins. Rare cases of the disease with non-vertical transmission have now been found to overlap with localized autosomal recessive hypotrichosis. The underlying gene, desmoglein 4 (DSG4), belongs to the desmosomal cadherin superfamily and is also expressed in the cortex of the hair follicle. PMID- 16702973 TI - Changes in NMDA receptor subunits and interacting PSD proteins in dorsolateral prefrontal and anterior cingulate cortex indicate abnormal regional expression in schizophrenia. AB - Abnormal expression of the N-methyl-D-Aspartate (NMDA) receptor and its interacting molecules of the postsynaptic density (PSD) are thought to be involved in the pathophysiology of schizophrenia. Frontal regions of neocortex including dorsolateral prefrontal (DLPFC) and anterior cingulate cortex (ACC) are essential for cognitive and behavioral functions that are affected in schizophrenia. In this study, we have measured protein expression of two alternatively spliced isoforms of the NR1 subunit (NR1C2 and NR1C2') as well as expression of the NR2A-D subunits of the NMDA receptor in DLPFC and ACC in post mortem samples from elderly schizophrenic patients and a comparison group. We found significantly increased expression of NR1C2' but not of NR1C2 in ACC, suggesting altered NMDA receptor cell membrane expression in this cortical area. We did not find significant changes in the expression of either of the NR1 isoforms in DLPFC. We did not detect changes of any of the NR2 subunits studied in either cortical area. In addition, we studied expression of the NMDA interacting PSD molecules NF-L, SAP102, PSD-95 and PSD-93 in ACC and DLPFC at both transcriptional and translational levels. We found significant changes in the expression of NF-L in DLPFC, and PSD-95 and PSD-93 in ACC; increased transcript expression was associated with decreased protein expression, suggesting abnormal translation and/or accelerated protein degradation of these molecules in schizophrenia. Our findings suggest abnormal regional processing of the NMDA receptor and its associated PSD molecules, possibly involving transcription, translation, trafficking and protein stability in cortical areas in schizophrenia. PMID- 16702974 TI - MPZL1/PZR, a novel candidate predisposing schizophrenia in Han Chinese. AB - The MPZL1/PZR gene has been mapped to 1q23.3, located in close proximity to a recognized schizophrenia susceptibility locus. Recently, the MPZL1/PZR gene has been found to be significantly upregulated in schizophrenia brain tissue and to play an important role in cell signaling, thus indicating that MPZL1/PZR could be a potential schizophrenia marker. To test this hypothesis, we selected three single nucleotide polymorphisms (SNPs) for genotyping in 523 Han Chinese trios. We found that two individual SNPs were significant at the Bonferroni's corrected significance level P<0.017: rs3767444 (chi2=6.299, P=0.0121) and rs2051656 (chi2=9.856, P=0.0017). Haplotype transmission/disequilibrium tests revealed a significant association with the disease (global P-value=1.064 x 10(-6)), but no specific transmission distortions. Thus, we propose that the MPZL1/PZR gene may be important in the predisposition to schizophrenia among Han Chinese. PMID- 16702975 TI - Depressive symptoms and insulin resistance in young adult males: results from the Northern Finland 1966 birth cohort. AB - The association between insulin resistance (IR) and depression is a subject of growing research interest, especially as previous population-based studies have presented conflicting findings. The present study extends our understanding about the putative impact of the severity of depressive symptoms on this association and it provides further epidemiological evidence in support of earlier findings, suggesting that the association between IR and depression is present already in young adult males. To determine the impact of the severity of depressive symptoms on the putative association between IR and depression in young adult males, we were given access to the Northern Finland 1966 Birth Cohort database. During the 31-year follow-up survey of this genetically homogeneous birth cohort, IR was assessed by 'Qualitative Insulin Sensitivity Check Index' (QUICKI), and severity of depressive symptoms by 'Hopkins' Symptom Checklist-25' (HSCL-25). This study involved 2,609 male cohort members with complete variable information. In men, the means of the QUICKI-values decreased (i.e., IR increased) in line with the increased severity of depressive symptoms as assessed by HSCL-25 subgroups (analysis of covariance P-value for trend, P=0.003). In multivariate generalized logistic regression analyses, after adjusting for confounders, IR was positively associated with current severe depressive symptoms, the odds ratio (OR) being over threefold (adjusted OR 3.15, 95% confidence interval 1.48-6.68) and the value of OR increased in parallel with a tighter definition of IR (P-value for trend=0.007). The results indicate that in young males, a positive association exists specifically with severe depressive symptoms. PMID- 16702977 TI - Refining the attention deficit hyperactivity disorder phenotype for molecular genetic studies. AB - It is well established that attention deficit hyperactivity disorder (ADHD) is a familial and highly heritable disorder. Consequently, much effort is being directed towards searching for specific susceptibility genes. There is a growing trend, across the field of complex disease genetics, towards undertaking secondary analyses based on refined phenotypic definitions and in testing whether specific susceptibility genes modify the phenotypic presentation of the disorder in question. It is crucial that good, empirically derived arguments are made before undertaking multiple analyses on different phenotype refinements. In this review article, we consider the evidence from genetic epidemiological studies as well as key clinical studies that provide guidance on examining the ADHD phenotype for the purpose of molecular genetic studies. Specifically, findings on categorical versus dimensional conceptualisations of ADHD, reporter effects, comorbidity, ADHD subtypes and persistence are reviewed. Current evidence suggests that for the purpose of identifying susceptibility genes for ADHD, parent and teachers should be used as informants and that focusing on the clinical diagnosis of ADHD is useful. There is also good empirical support in favour of examining antisocial behaviour in ADHD. Genetic studies of dimensional ADHD are useful for other complementary purposes. PMID- 16702976 TI - Specificity and timing of neocortical transcriptome changes in response to BDNF gene ablation during embryogenesis or adulthood. AB - Brain-derived neurotrophic factor (BDNF) has been reported to be critical for the development of cortical inhibitory neurons. However, the effect of BDNF on the expression of transcripts whose protein products are involved in gamma amino butric acid (GABA) neurotransmission has not been assessed. In this study, gene expression profiling using oligonucleotide microarrays was performed in prefrontal cortical tissue from mice with inducible deletions of BDNF. Both embryonic and adulthood ablation of BDNF gave rise to many shared transcriptome changes. BDNF appeared to be required to maintain gene expression in the SST-NPY TAC1 subclass of GABA neurons, although the absence of BDNF did not alter their general phenotype as inhibitory neurons. Furthermore, we observed expression alterations in genes encoding early-immediate genes (ARC, EGR1, EGR2, FOS, DUSP1, DUSP6) and critical cellular signaling systems (CDKN1c, CCND2, CAMK1g, RGS4). These BDNF-dependent gene expression changes may illuminate the biological basis for transcriptome changes observed in certain human brain disorders. PMID- 16702979 TI - A 40-basepair VNTR polymorphism in the dopamine transporter (DAT1) gene and the rapid response to antidepressant treatment. AB - Finding predictors of the response to antidepressant therapy is a major goal of molecular psychiatry. The genes encoding the serotonin (SERT) and dopamine (DAT1) transporters are among the possible candidate genes modulating an individual's antidepressant response. In a naturalistic prospective cohort study with a total of 190 fully assessed patients, improvement of depression symptoms during the 3 weeks following initiation of antidepressant therapy was recorded using the 21 item Hamilton Depression Rating Scale (HDRS). The SLC6A3 3' UTR 40-bp variable number of tandem repeats (VNTR) and the SLC6A4 5' 44-bp insertion/deletion polymorphism were analyzed by polymerase chain reaction. There was a significantly smaller number of rapid responders among homozygous carriers of the DAT1 9-repeat allele (9/9) than among heterozygous (9/10) and homozygous (10/10) carriers of the 10-repeat allele (19 versus 37 versus 52%, respectively, P=0.0037). Median decline in HDRS score was 35, 40, and 52% in patients with the 9/9, 9/10, and 10/10 genotypes, respectively (P=0.013). The effect was found in all classes of medications (selective serotonin reuptake inhibitors (SSRIs), tricyclics, mirtazapine, venlafaxine) and statistically significant also within the subgroup of patients having received SSRIs. The serotonin promoter insertion/deletion genotype had no effect in the entire study group, but there was an insignificant trend of better response in the l/l and l/s carriers who received SSRIs or mirtazapine. In conclusion, the dopamine transporter VNTR polymorphism influenced rapid response to antidepressant therapy. Compared with homozygous carriers of the 10-repeat allele, carriers of the 9/10 genotype had an odds ratio (OR) calculated by logistic regression analysis of 1.6 (95% CI 0.8 3.2) and carriers of the 9/9 genotype had an OR of 6.0 (1.5-24.4) for no or poor response. Further studies are required to confirm this clinical association and to elucidate the underlying mechanisms. PMID- 16702978 TI - UBD, a downstream element of FOXP3, allows the identification of LGALS3, a new marker of human regulatory T cells. AB - Here, we report the identification of the ubiquitin-like gene UBD as a downstream element of FOXP3 in human activated regulatory CD4(+)CD25(hi) T cells (T(reg)). Retroviral transduction of UBD in human allo-reactive effector CD4(+) T helper (T(h)) cells upregulates CD25 and mediates downregulation of IL4 and IL5 expression similar to overexpression of FOXP3. Moreover, UBD impairs T(h) cell proliferation without upregulation of FOXP3 and impairs calcium mobilization. In the presence of ionomycin, overexpression of UBD in T(h) cells leads to the induction of IL1R2 that resemble FOXP3-transduced T(h) cells and naturally derived T(reg) cells. A comparison of the transcriptome of FOXP3- and UBD transduced T(h) cells with T(reg) cells allowed the identification of the gene LGALS3. However, high levels of LGALS3 protein expression were observed only in human CD4(+)CD25(hi) derived T(reg) cells and FOXP3-transduced T(h) cells, whereas little was induced in UBD-transduced T(h) cells. Thus, UBD contributes to the anergic phenotype of human regulatory T cells and acts downstream in FOXP3 induced regulatory signaling pathways, including regulation of LGALS3 expression. High levels of LGALS3 expression represent a FOXP3-signature of human antigen stimulated CD4(+)CD25(hi) derived regulatory T cells. PMID- 16702980 TI - Pharmacogenetic of antirheumatic treatments: clinical implications. AB - Preliminary pharmacogenetic data suggest that germline genetic informations might be of value in individualizing disease-modifying antirheumatic drugs (DMARDs) therapy in various autoimmune chronic inflammatory diseases. Either DMARDs small molecules (DMARDs-SM) or DMARDs biological therapies (DMARDs-BT) might be selected for their lower toxicity or better efficacy based on single-nucleotide polymorphisms (SNPs) of genes governing the metabolism of drugs, or the response of immune cells to proinflammatory molecules, or the proinflammatory molecular activity of immune cells. Data available for one DMARDs-SM, methotrexate, suggest that a careful assessment of the SNPs of four enzymes involved in the folate metabolism allow one to construct a genetic index of toxicity (toxicogenetic index) that might be employed in daily practice to find the patient's most at risk. Only the full knowledge of the various gene polymorphisms controlling the phenotypic manifestations of the inflammatory-immunological milieu of each rheumatic disease will allow one to obtain the clear definition of a personalized medicine. Few different cytokine gene SNPs seem to be of importance in determining the susceptibility to diseases, or the aggressiveness of diseases. The role of genetics in affecting a possible clinical response to DMARDs-BT targeting specific inflammatory molecules or their receptors still has to be defined. However, the available data suggest that cytokine (and/or receptors) gene SNPs might indeed play a role in determining the biological effects, hence the clinical effectiveness of DMARDs-BT. Crucial to this aim will be the prospective analysis of clinical benefits and safety on the basis of the at baseline stratification of gene SNPs in each chronic inflammatory rheumatic disease before starting any new DMARDs-SM or DMARDs-BT. PMID- 16702981 TI - Antihypertensive therapy, the alpha-adducin polymorphism, and cardiovascular disease in high-risk hypertensive persons: the Genetics of Hypertension Associated Treatment Study. AB - In a double-blind, outcome trial conducted in hypertensive patients randomized to chlorthalidone (C), amlodipine (A), lisinopril (L), or doxazosin (D), the alpha adducin Gly460Trp polymorphism was typed (n=36 913). Mean follow-up was 4.9 years. Relative risks (RRs) of chlorthalidone versus other treatments were compared between genotypes (Gly/Gly+Gly/Trp versus Trp/Trp). Primary outcome was coronary heart disease (CHD). Coronary heart disease incidence did not differ among treatments or genotypes nor was there any interaction between treatment and genotype (P=0.660). Subgroup analyses indicated that Trp allele carriers had greater CHD risk with C versus A+L in women (RR=1.31) but not men (RR=0.91) with no RR gender differences for non-carriers (gender-gene-treatment interaction, P=0.002). The alpha-adducin gene is not an important modifier of antihypertensive treatment on cardiovascular risk, but women Trp allele carriers may have increased CHD risk if treated with C versus A or L. This must be confirmed to have implications for hypertension treatment. PMID- 16702982 TI - The serotonin transporter gene polymorphism is not associated with smoking behavior. AB - Nicotine increases serotonin release in the brain and symptoms of nicotine withdrawal may be modulated by diminished serotonergic neurotransmission. The promoter region of the serotonin transporter gene, solute carrier family neurotransmitter transporter member 4 (SLC6A4), contains a functional tandem repeat polymorphism. The long (L) variant is more actively transcribed than the short (S) variant and is associated with a higher serotonin uptake. To investigate the potential role of this polymorphism for smoking behavior, SLC6A4 genotypes were determined in two different studies, the SMOKING GENES case control study (470 current smokers and 419 subjects who had never smoked) and the cross-sectional Ludwigshafen risk and cardiovascular health (LURIC) study (777 current smokers and 1178 subjects who had never smoked). In the SMOKING GENES case-control study, SLC6A4 genotype frequencies were not statistically different between smokers (LL: 30.9%; LS: 46.8%; SS: 16.4%) and non-smokers (LL: 36.3%; LS: 41.8%; SS: 14.3%; P=0.13). Similar results were obtained in the cross-sectional LURIC study (smokers: LL, 36.5%, LS, 45.6%, SS, 17.9%; non-smokers: LL, 33.6%, LS, 48.9%, SS, 17.6%; P=0.33). SLC6A4 genotypes were furthermore not associated with Fagerstrom Tolerance Questionnaire score, packyears, number of cigarettes smoked per day or previous attempts to quit smoking. We conclude that the SLC6A4 promoter polymorphism is not a major determinant of smoking behavior in Caucasian. PMID- 16702983 TI - First among equals: competition between genetically identical cells. AB - Competition between genetically identical organisms is considered insignificant in evolutionary theory because it is presumed to have little selective consequence. We argue that competition between genetically identical cells could improve the fitness of a multicellular organism by directing fitter cells to the germ line or by eliminating unfit cells, and that cell-competition mechanisms have been conserved in multicellular organisms. We propose that competition between genetically identical or highly similar units could have similar selective advantages at higher organizational levels, such as societies. PMID- 16702984 TI - Molecular markers and their prognostic impact in patients with advanced prostate cancer undergoing intermittent androgen suppression. AB - OBJECTIVE: Tumour features were evaluated during intermittent androgen suppression (IAS), and their prognostic impact on the first off-treatment time was analysed. PATIENTS AND METHODS: Twenty patients with advanced prostate cancer underwent three consecutive prostate biopsies during the first cycle, namely at the beginning of androgen deprivation, 8 months after continuous therapy and at the time of prostate-specific antigen (PSA) progression above 20 ng/ml. Biopsy specimens were immunohistochemically processed and analysed for the apoptotic index (AI), Ki-67, p53 and Bcl-2 to investigate eventual changes over time. Correlations and regression analysis were performed to assess the prognostic significance of clinical and pathological parameters in predicting the first off treatment time. RESULTS: In contrast to the AI, p53 and Bcl-2, Ki-67 was the only marker that significantly changed over time (P=0.008). The first off-treatment time correlated significantly with pretreatment PSA (r=-0.594; P<0.01), testosterone recovery time (r=0.590; P=0.013) and biopsy grade (r=-0.738; P<0.01); only the latter gaining an independent factor in the multivariate analysis (P=0.022). CONCLUSIONS: During IAS, Ki-67 was the only molecular marker that consistently changed over time. However, it did not correlate with off treatment time that was predicted independently by the initial biopsy grade only. First off-treatment time was best predicted by clinical parameters and molecular markers from needle biopsies did not further contribute to a better patient selection. PMID- 16702985 TI - Relevance of tumour necrosis factor-alpha for the inflammatory and nociceptive responses evoked by carrageenan in the mouse paw. AB - 1. The present study evaluated the participation of tumour necrosis factor-alpha (TNF-alpha) in the inflammatory and nociceptive responses evoked by carrageenan in the mouse paw. 2. The intraplantar injection of carrageenan (300 microg paw-1) induced a marked and biphasic paw oedema formation (peaks at 6 and 72 h), which was accompanied by a long-lasting mechanical allodynia (that remained elevated for up to 72 h) and a significant increase of myeloperoxidase (MPO) activity (peak at 6 h) in both Swiss and C57/BL6 mice. 3. The paw oedema, the elevation of MPO activity and to a lesser extent the mechanical allodynia elicited by carrageenan were found to be significantly reduced in TNF-alpha p55 receptor knockout mice. 4. Of interest, the systemic administration of an anti-TNF-alpha antibody produced a significant inhibition of paw oedema, mechanical allodynia and MPO activity. A noteworthy decrease in inflammatory and nociceptive responses caused by carrageenan was also observed when mice were previously treated with the preferential inhibitor of TNF-alpha synthesis, thalidomide. 5. The present results clearly indicate that the proinflammatory cytokine TNF-alpha plays a critical role in the oedema formation, as well as in the mechanical allodynia and the neutrophil migration, following carrageenan administration into the mouse paw. Intraplantar injection of carrageenan in mice could constitute a useful model for assessment of the in vivo effects of potential inhibitors of TNF-alpha related pathways. PMID- 16702986 TI - Cardioprotective effects of atrasentan, an endothelin-A receptor antagonist, but not of nitric oxide in diabetic mice with myocyte-specific overexpression of endothelial nitric oxide synthase. AB - 1. We investigated the roles of nitric oxide (NO) and endothelin-1 (ET-1) in organ dysfunction in diabetic mice with normal genotype (wild-type, WT) or myocyte-specific overexpression of endothelial NO synthase (eNOS) (transgenic, TG) after chronic oral treatment with the endothelin-A (ETA) receptor antagonist atrasentan. 2. Mice were rendered diabetic by injection of 200 mg kg-1 streptozotocin (STZ). Experimental groups were: untreated WT diabetic (n=9), untreated TG diabetic (n=9), atrasentan-treated WT diabetic (n=9), atrasentan treated TG diabetic (n=8) and the four corresponding nondiabetic groups (n=5). Atrasentan was administered orally via drinking water at 3 mg kg-1 per day over 28 days. All diabetic mice developed similar hyperglycaemia (27-30 mmol l-1). 3. Atrasentan treatment significantly improved left ventricular systolic and diastolic function in response to exogenous norepinephrine, but there were no differences between genotypes. 4. Atrasentan antagonized the diabetic impairments in endothelium-dependent coronary relaxation and thromboxane-receptor mediated aortic constriction. Further, it improved cardiac and renal oxidant status as evident from reduced tissue malondialdehyde levels. 5. Atrasentan reduced diabetic urine flow, proteinuria and plasma creatinine levels, but creatinine clearance was not significantly altered. 6. These results suggest that in experimental type 1 diabetes, blocking ETA receptors ameliorates myocardial, coronary and renal function and improves tissue oxidant status, whereas raising myocardial NO levels has neither beneficial nor deleterious effects on diabetic cardiomyopathy in this transgenic model. PMID- 16702987 TI - Pharmacological regulation of insulin secretion in MIN6 cells through the fatty acid receptor GPR40: identification of agonist and antagonist small molecules. AB - 1. Long chain fatty acids have recently been identified as agonists for the G protein-coupled receptors GPR40 and GPR120. Here, we present the first description of GW9508, a small-molecule agonist of the fatty acid receptors GPR40 and GPR120. In addition, we also describe the pharmacology of GW1100, a selective GPR40 antagonist. These molecules were used to further investigate the role of GPR40 in glucose-stimulated insulin secretion in the MIN6 mouse pancreatic beta cell line. 2. GW9508 and linoleic acid both stimulated intracellular Ca2+ mobilization in human embryonic kidney (HEK)293 cells expressing GPR40 (pEC50 values of 7.32+/-0.03 and 5.65+/-0.06, respectively) or GPR120 (pEC50 values of 5.46+/-0.09 and 5.89+/-0.04, respectively), but not in the parent HEK-293 cell line. 3. GW1100 dose dependently inhibited GPR40-mediated Ca2+ elevations stimulated by GW9508 and linoleic acid (pIC50 values of 5.99+/-0.03 and 5.99+/ 0.06, respectively). GW1100 had no effect on the GPR120-mediated stimulation of intracellular Ca2+ release produced by either GW9508 or linoleic acid. 4. GW9508 dose dependently potentiated glucose-stimulated insulin secretion in MIN6 cells, but not in primary rat or mouse islets. Furthermore, GW9508 was able to potentiate the KCl-mediated increase in insulin secretion in MIN6 cells. The effects of GW9508 on insulin secretion were reversed by GW1100, while linoleic acid-stimulated insulin secretion was partially attenuated by GW1100. 5. These results add further evidence to a link between GPR40 and the ability of fatty acids to acutely potentiate insulin secretion and demonstrate that small-molecule GPR40 agonists are glucose-sensitive insulin secretagogues. PMID- 16702988 TI - Differentiation of hippocampal stem cells into functional neurons: evolving our understanding of monoamine oxidase-A inhibition. AB - Depression affects many millions of people worldwide and much is still unknown with respect to the mode of action of antidepressant drugs. The hippocampus has been associated with many psychiatric disorders, including clinical depression. Recently, stem cells have also been shown to reside within discrete regions of the hippocampus and can differentiate under a variety of conditions into neural cells. In this issue, Chiou et al. have elegantly demonstrated that cells isolated from the rat hippocampus, and treated with the antidepressant moclobemide, may be differentiated in vitro into neural cells exhibiting features to those of serotoninergic neurons. They have also suggested that this process was mediated, in part, through the expression of specific antiapoptotic genes (Bcl-2 and Bcl-XL) and via activation of extracellular-regulated kinase. This work raises the attractive possibility that the use of antidepressants, such as moclobemide, may exert neuroprotective and potentially neurogenerative effects not just in vitro, but also in vivo, through the selected differentiation of stem cells into functional neurons. The exact mechanisms by which such antidepressants differentiate neural stem cells still remains to be fully elucidated. PMID- 16702989 TI - Excitatory P2-receptors at sympathetic axon terminals: role in temperature control of cutaneous blood flow. AB - The mechanisms underlying the reduction in cutaneous blood flow in response to cooling are only partially understood. A study published in this issue of the British Journal of Pharmacology now provides evidence for the involvement of excitatory P2-receptors located at sympathetic axon terminals in the cooling induced vasoconstriction in the skin. Cooling appears to cause the release of adenine nucleotides followed by the activation of excitatory presynaptic P2 receptors at noradrenergic axon terminals. Activation of these excitatory P2 receptors induces the release of noradrenaline, which subsequently causes constriction of blood vessels in the skin by action on smooth muscle alpha(1)- and alpha(2)-adrenoceptors. The commentary discusses the implication of the results and remaining questions. PMID- 16702990 TI - Moclobemide upregulated Bcl-2 expression and induced neural stem cell differentiation into serotoninergic neuron via extracellular-regulated kinase pathway. AB - 1. Moclobemide (MB) is an antidepressant drug that selectively and reversibly inhibits monoamine oxidase-A. Recent studies have revealed that antidepressant drugs possess the characters of potent growth-promoting factors for the development of neurogenesis and improve the survival rate of serotonin (5 hydroxytrytamine; 5-HT) neurons. However, whether MB comprises neuroprotection effects or modulates the proliferation of neural stem cells (NSCs) needs to be elucidated. 2. In this study, firstly, we used the MTT (3-(4,5-dimethylthiazol-2 yl)-2,5-diphenyltetrazolium bromide) assay to demonstrate that 50 microM MB can increase the cell viability of NSCs. The result of real-time reverse transcription-polymerase chain reaction (RT-PCR) showed that the induction of MB can upregulate the gene expressions of Bcl-2 and Bcl-xL. By using caspases 8 and 3, ELISA and terminal dUTP nick-end labeling (TUNEL) assay, our data further confirmed that 50 microM MB-treated NSCs can prevent FasL-induced apoptosis. 3. The morphological findings also supported the evidence that MB can facilitate the dendritic development and increase the neurite expansion of NSCs. Moreover, we found that MB treatment increased the expression of Bcl-2 in NSCs through activating the extracellular-regulated kinase (ERK) phosphorylation. 4. By using the triple-staining immunofluorescent study, the percentages of serotonin- and MAP-2-positive cells in the day 7 culture of MB-treated NSCs were significantly increased (P<0.01). Furthermore, our data supported that MB treatment increased functional production of serotonin in NSCs via the modulation of ERK1/2. In sum, the study results support that MB can upregulate Bcl-2 expression and induce the differentiation of NSCs into serotoninergic neuron via ERK pathway. PMID- 16702991 TI - Local regulation of skin blood flow during cooling involving presynaptic P2 purinoceptors in rats. AB - 1. This study investigated a local effect of cooling on the plantar skin blood flow (PSBF) of tetrodotoxin-treated rats by laser-Doppler flowmetry. 2. When the air temperature around the left foot was locally cooled from 25 to 10 degrees C, the PSBF of the left foot decreased. 3. The response was inhibited by the alpha adrenoceptor antagonist phentolamine, the alpha1-adrenoceptor antagonist bunazosin, the alpha2-adrenoceptor antagonist RS79948, and bretylium and guanethidine that inhibit noradrenaline release from sympathetic nerves. Adrenalectomy of the rats did not affect the cooling-induced response. 4. The P2 purinoceptor antagonists suramin and PPADS also significantly suppressed the cooling-induced reduction of PSBF. However, the inhibitory effect of PPADS on the cooling-induced response was abolished after the treatment with phentolamine. Intra-arterial injections of ATPgammaS, a stable P2 purinoceptor agonist, at 25 degrees C caused a transient decrease in PSBF in a dose-dependent manner, which was significantly inhibited by phentolamine and guanethidine. 5. These results suggest a novel mechanism for local cooling-induced reduction of skin blood flow in vivo; moderate cooling of the skin induces the release of ATP, which stimulates presynaptic P2 purinoceptors on sympathetic nerve terminals and facilitates the release of noradrenaline, thereby causing contractions of skin blood vessels via the activation of alpha1-and alpha2-adrenoceptors. PMID- 16702993 TI - Hypothalamic neuropeptide Y (NPY) and the attenuation of hyperphagia in streptozotocin diabetic rats treated with dopamine D1/D2 agonists. AB - 1. Dopamine is an appetite suppressant, while neuropeptide Y (NPY), an appetite stimulant in the brain, is reported to be involved in anorectic action induced by a combined administration of D1/D2 agonists in normal rats. In diabetic rats, however, these factors have not been studied. 2. Rats (including normal, diabetic and insulin-treated diabetic rats) were given daily injections of saline or D1/D2 agonists for 6 days. Changes in food intake and hypothalamic NPY content of these rats were assessed and compared. 3. The D1/D2 agonist-induced anorectic responses were altered in diabetic rats compared to normal rats treated similarly. Both the anorectic response on the first day of dosing and the tolerant response on the subsequent days were attenuated. 4. This alteration was independent of the neuroendocrine disturbance on feeding behavior since the basic pattern of food intake during the time course of a 24-h day/night cycle was similar in normal and diabetic rats; the decrease of food intake following drug treatment was only shown at the initial interval of 0-6 h in both groups of rats. 5. However, this alteration coincided with changes in NPY content following D1/D2 coadministration. The replacement of insulin in diabetic rats could normalize both NPY content and D1/D2 agonist-induced anorexia. 6. It is demonstrated that the response of D1/D2 agonist-induced appetite suppression is attenuated in diabetic rats compared to normal rats and that elevated hypothalamic NPY content may contribute to this alteration. PMID- 16702992 TI - Cholesteryl butyrate solid lipid nanoparticles inhibit adhesion of human neutrophils to endothelial cells. AB - 1. Adhesion of polymorphonuclear cells (PMNs) to vascular endothelial cells (EC) is a critical step in recruitment and infiltration of leukocytes into tissues during inflammation. High doses of butyric acid have been shown to ameliorate inflammation in inflammatory bowel diseases (IBD). Cholesteryl-butyrate solid lipid nanoparticles (chol-but SLN) as prodrug are a possible delivery system for butyric acid. 2. Sodium butyrate or chol-but SLN were coincubated with human PMNs and human umbilical vein EC (HUVEC); adhesion was quantified by computerized microimaging fluorescence analysis. Both chol-but SLN and sodium butyrate displayed antiadhesive effects on FMLP- and IL-1beta-stimulated cells in a concentration-response curve (10(-8)-10(-5) M), but chol-but SLN were in all cases more active. Moreover, chol-but SLN inhibited FMLP-induced adhesion of PMNs to FCS-coated plastic wells, thus showing a direct effect on PMNs, while sodium butyrate had little effect. Confocal microscopy showed that fluorescent SLN entered PMNs and HUVEC after 10 min incubation. Chol-but SLN acted either on activated PMN or HUVEC. 3. Chol-but SLN inhibited O2-* production and myeloperoxidase release by PMNs evoked by FMLP, in a dose-dependent, but not time dependent, manner and were more active than sodium butyrate. 4. In conclusion, in all tests chol-but SLN were more active than sodium butyrate. Thus, chol-but SLN might be a valid alternative to sodium butyrate in the anti-inflammatory therapy of ulcerative colitis, avoiding complications related to the administration of sodium butyrate. PMID- 16702994 TI - Characterization and effects on cAMP accumulation of adrenomedullin and calcitonin gene-related peptide (CGRP) receptors in dissociated rat spinal cord cell culture. AB - Adrenomedullin (AM) and calcitonin gene-related peptide (CGRP) have structural similarities, interact with each others receptors (calcitonin receptor-like receptor (CLR)/receptor-activity-modifying proteins (RAMPs)) and show overlapping biological activities. AM and CGRP receptors are chiefly coupled to cAMP production. In this study, a method of primary dissociated cell culture was used to investigate the presence of AM and CGRP receptors and their effects on cAMP production in embryonic spinal cord cells. Both neuronal and non-neuronal CLR immunopositive cells were present in our model. High affinity, specific [(125)I] AM binding sites (K(d) 79 +/- 9 pM and B(max) 571 +/- 34 fmol mg(-1) protein) were more abundant than specific [(125)I]-CGRP binding sites (K(d) 12 +/- 0.7 pM and B(max) 32 +/- 2 fmol mg(-1) protein) in embryonic spinal cord cells. Specific [(125)I]-AM binding was competed by related molecules with a ligand selectivity profile of rAM > hAM(22-52) > rCGRPalpha > CGRP(8-37) >> [r-(r(*),s(*))]-N-[2-[[5 amino-1-[[4-(4-pyridinyl)-1-piperazinyl]carbonyl]pentyl]amino]-1-[(3,5-dibromo-4 hydroxyphenyl)methyl]-2-oxoethyl]-4-(1,4-dihydro-2-oxo-3(2H)-quinazolinyl)-,1 piperidinecarboxamide (BIBN4096BS). Specific [(125)I]-CGRP binding was competed by rCGRPalpha > rAM > or = CGRP(8-37) > or = BIBN4096BS > hAM(22-52). Cellular levels of cAMP were increased by AM (pEC(50) 10.2 +/- 0.2) and less potently by rCGRPalpha (pEC(50) 8.9 +/- 0.4). rCGRPalpha-induced cAMP accumulation was effectively inhibited by CGRP(8-37) (pA(2) 7.63 +/- 0.44) and hAM(22-52) (pA(2) 6.18 +/- 0.21) while AM-stimulation of cAMP levels was inhibited by CGRP(8-37) (pA(2) 7.41+/- 0.15) and AM(22-52) (pA(2) 7.26 +/- 0.18). BIBN4096BS only antagonized the effects of CGRP (pA(2) 8.40 +/- 0.30) on cAMP accumulation. These pharmacological profiles suggest that effects of CGRP are mediated by the CGRP(1) (CLR/RAMP1) receptor in our model while those of AM are related to the activation of the AM(1) (CLR/RAMP2) receptor subtype. PMID- 16702995 TI - Changes in vascular reactivity following administration of isoproterenol for 1 week: a role for endothelial modulation. AB - 1. The aim of this study was to assess the effects of treatment with isoproterenol (ISO, 0.3 mg kg-1 day-1, s.c.) for 7 days on the vascular reactivity of rat-isolated aortic rings. Additionally, potential mechanisms underlying the changes that involved the endothelial modulation of contractility were investigated. 2. Treatment with ISO induced cardiac hypertrophy without changes in haemodynamic parameters. Aortic rings from ISO-treated rats showed an increase in the contraction response to phenylephrine (PHE) and serotonin, but did not change relaxations produced by acetylcholine or isoproterenol. Removal of the endothelium increased the responses to PHE in both groups. However, this procedure was less effective in ISO-treated as compared with control rats. Endothelial cell removal abolished the increase in the response to PHE in ISO treated rats. The presence of Nomega-nitro-L-arginine methyl ester shifted the concentration-response curve to PHE to the left in both groups of rats. However, this effect was more pronounced in the ISO group. In addition, aminoguanidine (50 microM) potentiated the actions of PHE only in the ISO group. ISO treatment increased nitric oxide synthase (NOS) activity and neuronal NOS and endothelial NOS protein expression in the aorta. 3. Neither losartan (10 microM) nor indomethacin (10 microM) abolished the effects of ISO on the actions of PHE. Superoxide dismutase (SOD, 150 U ml-1) and L-arginine (5 mM), but neither catalase (300 U ml-1) nor apocynin (100 microM), blocked the effect of ISO treatment. In addition, we observed an increase in superoxide anion levels as measured by ethidium bromide fluorescence and of copper and zinc superoxide dismutase protein expression in ISO-treated rats. 4. In conclusion, our data suggest that ISO treatment alters the endothelial cell-mediated modulation of the contraction to PHE in rat aorta. The increased maximal response of PHE seems to be due to an increase in superoxide anion generation, which inactivates some of the basal NO produced and counteracts NO-mediated negative modulation even in the presence of high NO production and antioxidant defence. PMID- 16702996 TI - Interaction between cyclodextrin and neuronal membrane results in modulation of GABA(A) receptor conformational transitions. AB - Cyclodextrins (CDs) are nanostructures widely applied in biotechnology and chemistry. Owing to partially hydrophobic character, CDs interact with biological membranes. While the mechanisms of CDs interactions with lipids were widely studied, their effects on proteins are less understood. In the present study we investigated the effects of beta cyclodextrin (betaCD) on GABA(A) receptor (GABA(A)R) gating. To reliably resolve the kinetics of conformational transitions, currents were elicited by ultrafast gamma-aminobutyric acid (GABA) applications to outside-out patches from rat cultured hippocampal neurons. betaCD increased the amplitude of responses to saturating GABA concentration ([GABA]) in a dose-dependent manner and this effect was accompanied by profound alterations in the current kinetics. Current deactivation was slowed down by betaCD but this effect was biphasic with a maximum at around 0.5 mM betaCD. While the fast deactivation time constant was monotonically slowed down within considered betaCD concentration range, the slow component first increased and then, at millimolar betaCD concentration, decreased. The rate and extent of desensitization was decreased by betaCD in a dose-dependent manner. The analysis of current responses to nonsaturating [GABA] indicated that betaCD affected the GABA(A)R agonist binding site by slowing down the unbinding rate. Modulation of GABA(A)R desensitization and binding showed different concentration-dependence suggesting different modualtory sites with higher affinity of the latter one. All the betaCD effects were fully reversible indicating that cholesterol uptake into betaCD was not the primary mechanism. We conclude that betaCD is a strong modulator of GABA(A)R conformational transitions. PMID- 16702998 TI - Pancreatic polypeptide in obese children before and after weight loss. AB - OBJECTIVE: Little is known concerning pancreatic polypeptide (PP) in weight loss and in childhood obesity. METHODS: Fasting PP, leptin and insulin concentrations were determined in 38 obese children and compared with 35 lean children of the same age, gender and pubertal stage. Furthermore, changes of PP concentrations over a 1-year period were analyzed in the obese children participating in a weight loss intervention program. RESULTS: Obese children had significantly (P<0.01) lower PP, and higher leptin and insulin levels compared to lean children. In multiple linear regression analysis, PP was significantly negatively correlated to body mass index (P<0.01), but not to leptin, insulin, age, gender and pubertal stage. Changes of PP did not significantly correlate to changes of insulin (r=0.07, P=0.343) and leptin (r=-0.02, P=0.459). The substantial weight loss in 17 children led to a significant (P<0.05) increase in PP and decrease in insulin and leptin. In the 21 children without substantial weight loss, there were no significant changes in PP, insulin and leptin. CONCLUSIONS: PP concentrations are decreased in obese children and independent of age, gender, pubertal stage, leptin and insulin. The decrease of PP in obese children normalized after weight loss. Therefore, low PP concentrations reflect the overweight status, rather than cause it. PMID- 16702997 TI - Mechanism of action of novel NO-releasing furoxan derivatives of aspirin in human platelets. AB - Incorporation of a nitric oxide (NO)-releasing moiety in aspirin can overcome its gastric side effects. We investigated the NO-release patterns and antiplatelet effects of novel furoxan derivatives of aspirin (B8 and B7) in comparison to existing antiplatelet agents. Cyclooxygenase (COX) activity was investigated in purified enzyme using an electron paramagnetic resonance-based technique. Concentration-response curves for antiplatelet agents +/- the soluble guanylate cyclase inhibitor, ODQ (50 microM) were generated in platelet-rich plasma (PRP) and washed platelets (WP) activated with collagen using turbidometric aggregometry. NO was detected using an isolated NO electrode. The furoxan derivatives of aspirin (B8, B7) and their NO-free furazan equivalents (B16, B15; all 100 microM) significantly inhibited COX activity (P < 0.01; n = 6) in vitro and caused aspirin-independent, cGMP-dependent inhibition of collagen-induced platelet aggregation in WP. B8 was more potent than B7 (PRP IC(50) = 0.62 +/- 0.1 microM for B8; 400 +/- 89 microM for B7; P < 0.0001. WP IC(50)s = 0.6 +/- 0.1 and 62 +/- 10 microM, respectively). The NO-free furazan counterparts were less potent antiplatelet agents (WP IC(50)s = 54 +/- 3 microM and 62 +/- 10 microM, respectively; P < 0.0001, B8 vs B16). Of the hybrids investigated, only B8 retained antiplatelet activity in PRP.NO release from furoxan-aspirin hybrids was undetectable in buffer alone, but was accelerated in the presence of either plasma or plasma components, albumin (4%), glutathione (GSH; 3 microM) and ascorbate (50 microM), the effects of which were additive for B7 but not B8. NO generation from furoxans was greatly enhanced by platelet extract, an effect that could largely be explained by the synergistic effect of intracellular concentrations of GSH (3 mM) and ascorbate (1 mM). We conclude that the decomposition of furoxan-aspirin hybrids to generate biologically active NO is catalysed by endogenous agents which may instil a potential for primarily intracellular delivery of NO. The blunting of the aspirin effects of furoxan hybrids is likely to be due to loss of the acetyl moiety in plasma; the observed antiplatelet effects are thereby primarily mediated via NO release. Compounds of this class might represent a novel means of inhibiting platelet aggregation by a combination of NO generation and COX inhibition. PMID- 16702999 TI - Functional characterization of human NTRK2 mutations identified in patients with severe early-onset obesity. AB - OBJECTIVE: The neurotrophin receptor TrkB has been implicated in the regulation of energy homeostasis in rodents. We have previously identified four rare missense mutations in the gene encoding TrkB, NTRK2, in 198 severely obese children with developmental delay. We have now undertaken a more detailed analysis of the in vitro functional consequences of the mutations identified: I98V, P660L, T821A and Y722C. DESIGN: Wild-type and mutant TrkB receptor constructs were stably transfected into PC12 cells and the signaling responses to the endogenous ligand, brain-derived neurotrophic factor (BDNF), were examined by Western blotting of cell lysates. In the case of Y722C, PC12 cells stably expressing this mutant were studied for their ability to respond to BDNF by promoting neurite outgrowth and cell survival. RESULTS: Further functional characterization of the previously reported Y722C TrkB mutation reveals impaired activation of mitogen-activated protein kinase, phospholipase C-gamma and Akt, as well as reduced BDNF-induced neurite outgrowth and cell survival in stably transfected PC12 cell lines. However, the signaling properties of I98V, P660L and T821A were all indistinguishable from wild type. CONCLUSION: We provide further evidence for the impairment in signaling by Y722C and show that as well as a loss of signaling, this mutation affects the ability of TrkB to promote neurite outgrowth in response to BDNF. Thus, impaired hypothalamic neurogenesis may contribute to the severe hyperphagia and obesity seen in the individual harboring the Y722C variant. The other three rare TrkB variants do not show reduced autophosphorylation or impaired downstream signaling in vitro and, as yet, it is unclear whether these variants contribute to obesity in these patients. PMID- 16703000 TI - Angiotensin II: a hormone that affects lipid metabolism in adipose tissue. AB - BACKGROUND: Alterations in adipose tissue lipolysis may contribute to the pathophysiology of obesity and insulin resistance. We examined the effects of angiotensin II (Ang II) on abdominal subcutaneous adipose tissue lipolysis in humans. METHODS AND RESULTS: First, adipocytes obtained from nine normal weight and seven obese subjects were stimulated with Ang II (10(-14)-10(-6) M). Glycerol concentration in the medium, used as an indicator of adipocyte lipolysis, was significantly reduced (approximately 20%) after Ang II stimulation in adipocytes from normal weight (P=0.04) and obese subjects (P<0.001). Based on these observations, adipocytes of seven additional obese subjects were stimulated with lower doses of Ang II (10(-17)-10(-6) M) in the presence and absence of Ang II type 1 (AT(1)) receptor blockade. Lipolysis was dose dependently inhibited by approximately 20 to 25% after Ang II stimulation (P=0.001). AT(1) receptor blockade completely abolished the Ang II-induced effects (P=0.35). CONCLUSION: Ang II directly inhibits abdominal subcutaneous adipocyte lipolysis in normal weight and obese subjects via the AT(1) receptor. PMID- 16703001 TI - Health-related quality of life, BMI and physical activity among US adults (>/=18 years): National Physical Activity and Weight Loss Survey, 2002. AB - OBJECTIVE: To examine the association between health-related quality of life (HRQOL) and physical activity (PA). METHODS: Cross-sectional data were obtained via a national telephone survey from 9173 respondents (30.9% response rate; 51.4% cooperation rate). Four indicators of HRQOL were measured: self-rated health, physically unhealthy days, mentally unhealthy days and activity limitation days. Prevalence estimates were calculated by body mass index (BMI) category and PA level. Logistic regression evaluated BMI as an effect modifier of the relationship between HRQOL and PA. RESULTS: Inactive adults reported more fair to poor HRQOL than active adults, regardless of BMI category (P<0.001). BMI did not modify the association between PA and any of the four HRQOL indicators. CONCLUSION: Prevalence of low HRQOL is inversely related to PA participation, and the relationship is not altered by BMI status. Regardless of their weight status, adults should be encouraged to engage in PA. PMID- 16703002 TI - Association between indices of body mass and antibody titres to heat-shock protein-60, -65 and -70 in healthy Caucasians. AB - We have previously shown that antibody titres to several heat-shock proteins (Hsps) are elevated in dyslipidaemic patients and subjects with established vascular disease. Obesity is known to be associated with raised serum inflammatory markers suggesting a state of heightened immune activation. Hence, we have investigated the association between indices of obesity and several Hsp antibody titres in healthy subjects. Subjects (n=170) were recruited from among employees at the University of Surrey and the Royal Surrey County Hospital, Guildford, UK. Of these subjects, 35 were obese with a body mass index (BMI)>/=30 kg/m(2) (19 male and 16 female subjects), 58 were overweight with 30>BMI>/=25 kg/m(2) (36 male and 22 female subjects) and 77 were of a normal weight with BMI<25 kg/m(2) (31 male and 46 female subjects). Overall, obese subjects had significantly higher plasma anti-Hsp-60 (P<0.001), anti-Hsp-65 (P<0.05) and anti Hsp-70 (P<0.05) compared with overweight and normal weight subjects. PMID- 16703003 TI - Relationships between glycaemic abnormalities, obesity and insulin resistance in nondiabetic Polynesians of New Caledonia. AB - OBJECTIVE: Polynesians in New Caledonia have an increased risk for developing diabetes, compared to Melanesians or Europeans. They are also more prone to obesity. The aim of this study was to analyse differences in the pre-diabetic state that may explain the varying susceptibility to diabetes between these three ethnic groups, focusing on the balance between insulin resistance and capacity of pancreatic cells to secrete insulin. DESIGN AND SUBJECTS: The CALDIA Study is a population-based cross-sectional survey of diabetes prevalence conducted in New Caledonia. All participants who did not have diabetes, according to the results of a 0-2 h oral glucose tolerance test (n=392), were selected for analysis. RESULTS: Compared to Europeans, Polynesians and Melanesians had significantly higher body mass indices (BMI) and waist-to-hip ratios (WHRs). Polynesians had higher fasting plasma glucose values than Europeans or Melanesians (6.03 mmol/l, vs 5.78 and 5.46, respectively; P<0.0001). Fasting plasma insulin level and the estimate of insulin resistance by homeostasis model assessment were not significantly different between the three ethnic groups. Homeostasis model assessment estimate of beta-cell secretory capacity was lower in Polynesians compared to the two other ethnic groups (83.1 mU/mmol, vs 119.3 and 125.2, respectively; P<0.02). CONCLUSION: Despite a high prevalence of central obesity, as judged by high BMI and WHR, in Polynesians of New Caledonia, their high risk of diabetes may be more strongly related to a defect in insulin secretion capacity than to insulin resistance. PMID- 16703004 TI - Efficacy and safety of topiramate in combination with metformin in the treatment of obese subjects with type 2 diabetes: a randomized, double-blind, placebo controlled study. AB - OBJECTIVE: To investigate the efficacy and safety of topiramate in obese subjects with type 2 diabetes treated with metformin. DESIGN: This was a multicenter, double-blind, placebo-controlled trial. All subjects received a non pharmacological program of diet, exercise and behavioral modification throughout the study; the assigned diet was 600 kcal/day less than the subject's individually calculated energy expenditure. After a 6-week single-blind placebo run-in, subjects were randomized to placebo, topiramate 96 mg/day or topiramate 192 mg/day. Following an 8-week titration period, subjects remained on their assigned dose for 52 weeks. However, the sponsor ended the study early in order to develop a new controlled-release formulation with the potential to enhance tolerability and simplify dosing in this patient population. A total of 646 obese men and women (age: 18-75 years, body mass index: 27-50 kg/m(2)) with an established history of type 2 diabetes mellitus controlled by metformin monotherapy were randomized. Efficacy was assessed in a pre-determined modified intent-to-treat (MITT) population of 307 subjects whose randomization date would have allowed them to complete 24 weeks on study medication before the announcement of study termination. MEASUREMENTS: Joint primary efficacy parameters were mean percent change in weight and change in glycosylated hemoglobin (HbA(1c)) from baseline to week 24. RESULTS: Subjects in the placebo, topiramate 96 mg/day and topiramate 192 mg/day groups lost 1.7%, 4.5% (P<0.001) and 6.5% (P<0.001), respectively, of their baseline body weight and had absolute decreases in HbA(1c) of 0.1%, 0.4% (P<0.001) and 0.6% (P<0.001) (MITT, last observation carried forward). Topiramate-treated subjects also experienced statistically significant decreases in systolic blood pressure. Most common adverse events were paresthesia and events related to the central nervous system. CONCLUSIONS: Topiramate was effective for weight reduction and improvement in glycemic control in obese subjects with type 2 diabetes treated with metformin monotherapy. Further study in obese diabetics is warranted. PMID- 16703006 TI - Holiday snaps. PMID- 16703005 TI - Adipogenic human adenovirus-36 reduces leptin expression and secretion and increases glucose uptake by fat cells. AB - OBJECTIVE: Human adenovirus Ad-36 causes adiposity in animal models and enhances differentiation and lipid accumulation in human and 3T3-L1 preadipocytes, which may, in part, explain the adipogenic effect of Ad-36. We determined the consequences of Ad-36 infection on leptin and glucose metabolism in fat cells. DESIGN: 3T3-L1 preadipocytes were used to determine the effect of infection by human adenoviruses Ad-36, Ad-2, Ad-9 and Ad-37 on leptin secretion and lipid accumulation. Rat primary adipocytes were used to determine the effect of Ad-36 infection on leptin secretion and glucose uptake in vitro. Furthermore, the effect of Ad-36 on expressions of leptin and selected genes of de novo lipogenesis pathway of visceral adipose tissue were compared ex vivo, between Ad 36 infected and uninfected control rats. RESULTS: Ad-36 suppressed the expression of leptin mRNA in 3T3-L1 cells by approximately 58 and 52% on days 3 and 5 post infection, respectively. Leptin release normalized to cellular lipid content was 51% lower (P<0.002) in the Ad-36 infected 3T3-L1 cells. Lipid accumulation was significantly greater and leptin secretion was lower for the 3T3-L1 cells infected with other human adenoviruses Ad-9, Ad-36, or Ad-37. Whereas, human adenovirus Ad-2 did not influence cellular lipid accumulation or the leptin release. In rat primary adipocytes, Ad-36 reduced leptin release by about 40% in presence of 0.48 (P<0.01) or 1.6 nM insulin (P<0.05) and increased glucose uptake by 93% (P<0.001) or 18% (P<0.05) in presence of 0 or 0.48 nM insulin, respectively. Next, the adipose tissue of Ad-36 infected rats showed two to fivefold lower leptin mRNA expression, and 1.6- to 21-fold greater expressions for acetyl Co-A carboxylase-1 and 1.2- to 6.3-fold greater expressions for fatty acid synthase, key genes of de novo lipogenesis, compared to the uninfected weight and adiposity matched controls. CONCLUSION: The in vitro and ex vivo studies show that Ad-36 modulates adipocyte differentiation, leptin production and glucose metabolism. Whether such a modulation contributes to enhanced adipogenesis and consequent adiposity in Ad-36 infected animals or humans needs to be determined. PMID- 16703007 TI - DIY dentistry. PMID- 16703008 TI - Don't be put off. PMID- 16703009 TI - Verbal dilemma. PMID- 16703010 TI - Ethical marketing. PMID- 16703012 TI - Bailing out of academia. PMID- 16703011 TI - Unseen evidence. PMID- 16703013 TI - A bit fishy. PMID- 16703014 TI - 3D anatomy online. PMID- 16703015 TI - Dubious diagnosis. PMID- 16703016 TI - Farce and fiction. PMID- 16703017 TI - The wrong journal. PMID- 16703018 TI - Academics: why has everyone left? PMID- 16703019 TI - Where has all the caries gone? PMID- 16703020 TI - 3D airway changes. PMID- 16703031 TI - Patterns of care and service use amongst children in the UK 2003. AB - BACKGROUND: The 2003 Children's Dental Health Survey is the fourth of the 10 yearly surveys of children's oral health in the United Kingdom. AIM: To detail the reported experience of dental services and dental treatment amongst children in the UK. METHOD: A self-completion questionnaire was distributed to a 50% sub sample of parents or carers of the children who were clinically examined in the 2003 UK Child Dental Health Survey. This included questions relating to parental and child experience of dental services and dental treatment. RESULTS: The proportion of UK five-year-olds reported as not having visited the dentist fell from 14% in 1983 to 6% in 2003 and the proportion reported as having visited the dentist before the age of two rose from 7% in 1983 to 31% in 2003. Over 80% of all children were reported to seek regular dental check-ups. Around 10% were reported to have had some difficulty in accessing NHS dental care while 5% of five-year-olds were reported to have experienced a general anaesthetic for dental procedures in 2003. Dental attendance was associated with social class and mothers' reported attendance patterns. CONCLUSIONS: In line with previously reported trends, the 2003 survey of children in the United Kingdom shows improvements in several areas but some aspects of attendance pattern continue to be associated with social class and mothers' attendance pattern. It is of concern that 10% of five-year-olds reported having experienced extractions and 5% general anaesthesia for dental treatment. PMID- 16703032 TI - How to reduce the stress of general dental practice: the need for research into the effectiveness of multifaceted interventions. AB - While the practice of dentistry has been demonstrated to be significantly stressful, there have been few published studies describing interventions to reduce the stress of dental practitioners. This article describes research into the prevention and alleviation of stress amongst a variety of healthcare professionals, including dental practitioners, and describes the findings from a small scale study of an intervention aimed at general dental practitioners who reported high levels of work related stress. It is argued that to be effective, interventions should be tailored to the individual needs of the practitioner, within a structured intervention framework. Further research into the effectiveness and cost-effectiveness of stress management for dental practitioners is required. PMID- 16703040 TI - A survey of oral and maxillofacial pathology specimens submitted by general dental practitioners over a 30-year period. AB - OBJECTIVES: To determine the range and frequency of diagnoses in specimens submitted for histopathological examination by general dental practitioners (GDPs). METHODS: A retrospective analysis was carried out of all cases submitted by GDPs for the period 1974-2003, using a Foxpro Windows database. The data were collated into 10 diagnostic categories each comprising number of diagnoses, percentage of each diagnosis within a diagnostic category and each diagnosis as a percentage of total cases. RESULTS: GDPs submitted 6,666 cases out of a total of 53,474 for this period. While the total number of specimens increased four-fold over the 30-year period, specimens from GDPs increased from 7% to 17%. The range of diagnoses increased from 18 to 45. Of the 617 GDPs who submitted material, 279 (45%) submitted less than two specimens each in 30 years. Nine malignant neoplasms were diagnosed. Other significant pathology included 320 benign neoplasms as well as diagnoses ranging from mucosal lesions such as lichen planus to odontogenic cysts. CONCLUSIONS: It is clear that GDPs have provided an increased number of biopsy specimens over the last three decades. This reflects an increasing demand by GDPs for a diagnostic oral histopathology service and their use of this service should be encouraged. PMID- 16703041 TI - A novel technique using preformed metal crowns for managing carious primary molars in general practice - a retrospective analysis. AB - BACKGROUND: There is a high level of untreated dental decay in primary teeth in Scotland. Despite evidence for the efficacy of preformed metal crowns (PMCs) for the restoration of primary molars, few are placed in general practice, possibly due to the interventive nature of the clinical procedure. There is, however, a novel way of placing PMCs involving no local anaesthesia, no caries removal and no preparation of the tooth: the Hall technique. AIM: To investigate the survival of carious primary teeth treated with PMCs placed using a novel, simplified method - the Hall technique. SETTING: General dental practice, in Scotland. METHOD: A retrospective analysis of practice records from one general practitioner, from 1988 to 2001. The majority of the 978 PMCs fitted on 259 children, using the Hall technique, were placed when there was clinical evidence of approximal caries into dentine. The Kaplan-Meier approach was used to analyse survival times and the Mantel-Haenszel Log rank test for comparison between tooth types. RESULTS: For all tooth types, the probability of surviving three years without being extracted or the PMC being lost, was 73.4% (95% confidence interval 70.1% to 76.4%), and for five years was 67.6% (95% confidence interval 63.3% to 71.5%). The probability of surviving without extraction alone for three years was 86.0% (95% confidence interval 83.2% to 88.4%), and for five years was 80.5% (95% confidence interval 76.5% to 83.9%). CONCLUSIONS: Hall technique restorations placed on primary molars with decay clinically into dentine, by a single operator in general dental practice, have a similar success rate to some other, more conventional, restorative techniques. The technique requires further evaluation through a prospective randomised control clinical trial before its use could be generally recommended. PMID- 16703042 TI - Revalidation of general dental practitioners in Scotland: the results of a pilot study. Part 2 - acceptability to practitioners. AB - AIM: To investigate the acceptablility of a pilot scheme of revalidation to general dental practitioners. METHOD: Ten general dental practitioners completed portfolios of evidence of being up to date and fit to practise. This portfolio was assessed by a panel of three experts, using an assessment tool developed specifically for that purpose. An action research methodology was used to evaluate participants' perceptions, consisting of a focus group and semi structured interviews. The views of the assessors on the portfolio and its assessment were collected using a questionnaire. RESULTS: The views of the participants on revalidation, the pilot scheme portfolio and its use, who should assess it and how its use could be supported were collected. Also areas of difficulty in using the portfolio were identified, along with suggestions for improving it and alternative ways of evidencing competence. Assessors noted that the quality of evidence was adequate, but also made suggestions for improvement of the portfolio. CONCLUSIONS: The pilot scheme appears to have been acceptable to the dentists in this scheme, given some caveats. The assessors felt that appraisal would significantly enhance any substantive scheme. PMID- 16703054 TI - Two implants for all edentulous mandibles. AB - Complete dentures have always been a poor substitute for natural teeth. Mandibular complete dentures frequently cause pain and discomfort, accelerated residual bone resorption, while failing to restore effective chewing. The provision of two implants to stabilise the mandibular complete denture can result in significant improvements. PMID- 16703055 TI - A victory for science and common sense. PMID- 16703057 TI - Higher moral ground. PMID- 16703056 TI - Drug management. PMID- 16703058 TI - Look at Sweden. PMID- 16703059 TI - Bisphosphonate guidelines. PMID- 16703060 TI - No evidence. PMID- 16703061 TI - Child abuse referral warning. PMID- 16703062 TI - Of relevance to dentistry. PMID- 16703063 TI - Disappointing research. PMID- 16703064 TI - Red wine mouthwash. PMID- 16703065 TI - How does it work? PMID- 16703066 TI - First class equivalent. PMID- 16703067 TI - No more assistants. PMID- 16703068 TI - Role models in academic dentistry. PMID- 16703081 TI - Oral health habits amongst children in the United Kingdom in 2003. AB - BACKGROUND: The 2003 Children's Dental Health Survey is the fourth in a series of decennial national children's dental health surveys. AIMS: This paper presents data on parental attitudes towards the care of children's teeth and gums, reported oral hygiene behaviours and clinical measures of hygiene and periodontal health in 2003 and highlights trends since previous surveys. METHOD: A total of 10,381 children were examined in schools by trained and calibrated examiners. Four measures of hygiene and periodontal health were recorded as part of the clinical examination. In addition, 3,342 questionnaires were completed by parents of a sub-sample of these children. The questionnaire enabled information to be collected about reported oral health behaviours and parental attitudes. RESULTS: About three quarters of children across all age groups are reported to brush their teeth twice daily. A range of oral health products was reported as being used in addition to toothbrushes and toothpaste. There is a trend in parental preferences towards restoration of teeth rather than extractions and towards a better understanding of dental caries prevention. Although overall a higher proportion of children in this survey appeared to have gingival inflammation, plaque and calculus than 10 and 20 years ago, there was no change in the proportion of older children with gingivitis. CONCLUSIONS: Dental practitioners have a role to play in reinforcing these positive attitudes and encouraging appropriate and effective oral hygiene behaviours in their child patients. PMID- 16703082 TI - An audit of referral practice for patients with impacted palatal canines and the impact of referral guidelines. AB - OBJECTIVE: To assess referral practice for patients presenting with impacted palatal maxillary canines, and to develop and evaluate the effectiveness of referral guidelines. DESIGN: Prospective clinical audit. SETTING: Southend and Basildon district general hospitals. SUBJECTS AND METHODS: The 'gold' standard was identified as regular dental attenders with unerupted palatal canines being referred by 12 years old, with a wait of no longer than 20 weeks from referral to assessment. Data were collected and compared to the defined standard. An algorithm outlining the correct management was developed and distributed to all local dentists. The cycle was repeated for a similar time period. RESULTS: Ninety eight per cent of patients were seen within 20 weeks during both cycles while the referrals increased from 85 to 109 patients. The percentage of patients referred by 12 years of age increased from 16.5% to 27% (p = 0.09). During the first cycle 82% of patients presented with retained deciduous maxillary canines. This was reduced to 76% during the second cycle (p = 0.29). CONCLUSION: Referral practice was generally poor when compared to the recommended good practice. More patients were referred after distribution of the guidelines, but the percentage referred by the recommended age was not statistically significantly improved. PMID- 16703089 TI - Dubious dataset. PMID- 16703090 TI - Underwhelming arguments. PMID- 16703091 TI - Sparing children pain. PMID- 16703092 TI - To fill or not to fill. PMID- 16703093 TI - A more comprehensive view. PMID- 16703094 TI - The development of a consensus among primary care dentists of referral criteria for school dental screening. AB - OBJECTIVE: To obtain consensus amongst a sample of primary care dentists in the North West of England on a set of clinical criteria that should trigger referral following school dental screening. DESIGN: Delphi process. SETTING: Primary dental care, England 2002. METHOD: Primary care dentists in the North West of England were randomly selected to complete a two round 'Delphi exercise' that included 10 potential referral criteria. The dentists were invited to express their level of support for the inclusion of each referral criterion. MAIN OUTCOME MEASURES: Level of agreement for each referral criterion. Acceptance of any criterion was that the interquartile range should be no more than 3 scale points with the lower value being no less than 7. RESULTS: Eighty-eight dentists, (72.7%), completed the Delphi exercise. Six referral criteria met with the groups' approval: Child with caries in permanent dentition. Child with darkened/discoloured permanent incisors. Child aged 9-10 years with overjet greater than 10 mm. Child over six years with either gross plaque, calculus or swollen gums. Child with evidence of sepsis. Child registered with a GDP with caries in permanent dentition. CONCLUSION: It is possible for a representative sample of primary care dentists in the North West to agree referral criteria following school dental screening. PMID- 16703095 TI - Assessing the quality of radiographic processing in general dental practice. AB - OBJECTIVES: To determine if a commercial device (Vischeck) for monitoring film processing quality was a practical option in general dental practice, and to assess processing quality among a group of GDPs in the West Midlands with this device. DESIGN: Clinical evaluation. SETTING: General dental practice, UK, 2004. METHOD: Ten GDP volunteers from a practice based research group processed Vischeck strips (a) when chemicals were changed, (b) one week later, and (c) immediately before the next change of chemicals. These were compared with strips processed under ideal conditions. Additionally, a series of duplicate radiographs were produced and processed together with Vischeck strips in progressively more dilute developer solutions to compare the change in radiograph quality assessed clinically with that derived from the Vischeck. RESULTS: The Vischeck strips suggested that at the time chosen for change of processing chemicals, eight dentists had been processing films well beyond the point indicated for replacement. Solutions were changed after a wide range of time periods and number of films processed. The calibration of the Vischeck strip correlated closely to a clinical assessment of acceptable film quality. CONCLUSIONS: Vischeck strips are a useful aid to monitoring processing quality in automatic developers in general dental practice. Most of this group of GDPs were using chemicals beyond the point at which diagnostic yield would be affected. PMID- 16703115 TI - An Investigation of Language Impairment in Autism: Implications for Genetic Subgroups. AB - Autism involves primary impairments in both language and communication, yet in recent years the main focus of research has been on the communicative deficits that define the population. The study reported in this paper investigated language functioning in a group of 89 children diagnosed with autism using the ADI-R, and meeting DSM-IV criteria. The children, who were between 4- and 14- years-old were administered a battery of standardized language tests tapping phonological, lexical, and higher-order language abilities. The main findings were that among the children with autism there was significant heterogeneity in their language skills, although across all the children, articulation skills were spared. Different subgroups of children with autism were identified on the basis on their performance on the language measures. Some children with autism have normal language skills; for other children, their language skills are significantly below age expectations. The profile of performance across the standardized measures for the language-impaired children with autism was similar to the profile that defines the disorder specific language impairment (or SLI). The implications of this language impaired subgroup in autism for understanding the genetics and definition of both autism and SLI are discussed. PMID- 16703114 TI - Genome sequence of Rickettsia bellii illuminates the role of amoebae in gene exchanges between intracellular pathogens. AB - The recently sequenced Rickettsia felis genome revealed an unexpected plasmid carrying several genes usually associated with DNA transfer, suggesting that ancestral rickettsiae might have been endowed with a conjugation apparatus. Here we present the genome sequence of Rickettsia bellii, the earliest diverging species of known rickettsiae. The 1,552,076 base pair-long chromosome does not exhibit the colinearity observed between other rickettsia genomes, and encodes a complete set of putative conjugal DNA transfer genes most similar to homologues found in Protochlamydia amoebophila UWE25, an obligate symbiont of amoebae. The genome exhibits many other genes highly similar to homologues in intracellular bacteria of amoebae. We sought and observed sex pili-like cell surface appendages for R. bellii. We also found that R. bellii very efficiently multiplies in the nucleus of eukaryotic cells and survives in the phagocytic amoeba, Acanthamoeba polyphaga. These results suggest that amoeba-like ancestral protozoa could have served as a genetic "melting pot" where the ancestors of rickettsiae and other bacteria promiscuously exchanged genes, eventually leading to their adaptation to the intracellular lifestyle within eukaryotic cells. PMID- 16703116 TI - Attachment, Friendship, and Psychosocial Functioning in Early Adolescence. AB - Fifth-graders' (N = 162; 93 girls) relationships with parents and friends were examined with respect to their main and interactive effects on psychosocial functioning. Participants reported on parental support, the quality of their best friendships, self-worth, and perceptions of social competence. Peers reported on aggression, shyness and withdrawal, and rejection and victimization. Mothers reported on psychological adjustment. Perceived parental support and friendship quality predicted higher global self-worth and social competence and less internalizing problems. Perceived parental support predicted fewer externalizing problems, and paternal (not maternal) support predicted lower rejection and victimization. Friendship quality predicted lower rejection and victimization for only girls. Having a supportive mother protected boys from the effects of low quality friendships on their perceived social competence. High friendship quality buffered the effects of low maternal support on girls' internalizing difficulties. PMID- 16703117 TI - The Effects of Parental Tobacco and Marijuana Use and Personality Attributes on Child Rearing in African-American and Puerto Rican Young Adults. AB - This study assessed the effect of the interrelationship of mothers' and fathers' tobacco and marijuana use with their personality attributes on some of their child rearing behaviors. We used a longitudinal design to analyze the data of 258 males and females who were seen four times over a 13-year period from early adolescence through young adult parenthood. Thirty-one percent of the multiple regression analyses revealed significant interactions between the effect of tobacco or marijuana use and a personality attribute on child rearing. The majority of these significant interactions suggested that protective personality characteristics were offset by substance use risks resulting in less adequate child rearing. If these results are substantiated in an experimental intervention, it suggests that having resilient personality attributes does not protect against the negative effects of tobacco or marijuana use on child rearing. PMID- 16703118 TI - Inhaled human insulin. AB - The benefit of subcutaneous insulin therapy in patients with diabetes is frequently limited due to difficulty in convincing patients of the importance of multiple daily insulin injections to cope effectively with meal-associated glycemic changes. Thus, the aim of achieving tight glycemic control, which is critical for reducing the risk of long-term diabetes-related complications, frequently remains elusive. The successful development of an inhalable insulin as a noninvasive alternative promises to change the management of diabetes. The first product to become available to patients is inhaled human insulin, a dry powder formulation packaged into discrete blisters containing 1 or 3 mg of dry powder human insulin and administered via a unique pulmonary inhaler device. It has recently been approved in both the United States and the European Union for the control of hyperglycemia in adult patients with type 1 or type 2 diabetes. The pharmacokinetic profile of inhaled human insulin closely mimics the natural pattern of insulin secretion, and resembles that of rapid-acting subcutaneous analogs. Similarly to rapid-acting subcutaneous analogs, inhaled human insulin has a more rapid onset of glucose-lowering activity compared to subcutaneous regular insulin, allowing it to be administered shortly before meals. It has a duration of glucose-lowering activity comparable to subcutaneous regular insulin and longer than rapid-acting insulin analogs. Inhaled human insulin effectively controls postprandial glucose concentrations in patients with type 1 or type 2 diabetes without increasing the risk of hypoglycemia, and even improves fasting glucose levels compared to subcutaneous insulin. Inhaled human insulin has an overall favorable safety profile. There are small reductions in lung function (1 1.5% of total lung forced expiratory volume in the first second [FEV1] capacity) after onset of treatment that are reversible in most patients if treatment is discontinued. Inhaled human insulin is associated with an increase in insulin antibody titers, especially in patients with type 1 diabetes. These increases are not associated with any clinical sequelae. Patient satisfaction data have shown that inhaled human insulin is associated with greater treatment satisfaction relative to subcutaneous insulin in patients with type 1 or type 2 diabetes. This review summarizes the current data on the clinical efficacy and safety of inhaled human insulin in patients with type 1 or type 2 diabetes. PMID- 16703119 TI - Argatroban: a direct thrombin inhibitor with reliable and predictable anticoagulant actions. AB - Pharmacological strategies aimed at the prevention of thrombotic complications are in continuous development. Argatroban is a synthetic small molecule derived from l-arginine with specific antithrombotic activity. Argatroban is a direct thrombin inhibitor that binds avidly and reversibly to the catalytic site of thrombin and that does not require other cofactors to exert its antithrombotic action. Due to its selective inhibitory mechanism, argatroban blocks both circulating and clot-bound thrombin. A rapid onset of its anticoagulant action is achieved after intravenous administration. The short elimination half-life of argatroban (52+/-16 minutes) ensures a rapid restoration of hemostasis upon cessation of treatment. Argatroban produces a predictable dose response, and its anticoagulant actions can be monitored easily through the routine coagulation tests activated partial thromboplastin time (aPTT) and activated clotting time (ACT). The specific mechanism of action and favorable pharmacokinetic profile of argatroban suggest that it could be beneficial in all indications where other intravenous anticoagulants are used. Results from clinical studies performed to date show that, when administered to patients with heparin-induced thrombocytopenia (HIT) or HIT with thrombosis (HITTS) in two large-scale, nonrandomized, prospective trials, argatroban reduced a combined endpoint of morbidity and mortality when compared with historical controls. Argatroban was well tolerated in clinical trials of patients with HIT and caused no increase in bleeding risk compared with historical controls. Argatroban does not induce the formation of antibodies that can neutralize its anticoagulant effect, prolong its half-life or enhance its activity. The U.S. Food and Drug Administration has approved the use of this drug as an alternative antithrombotic treatment for patients with HIT as well as for patients with or at risk for HIT undergoing percutaneous coronary interventions. In 2004 (Sweden), 2005 (Germany, the Netherlands, Austria and Iceland) and 2006 (Denmark) argatroban was approved for anticoagulation in adult patients with heparin-induced thrombocytopenia type II who require parenteral antithrombotic therapy. PMID- 16703120 TI - Cilomilast. AB - The central role of cyclic nucleotides as intracellular second messengers dates back almost 50 years. The importance of phosphodiesterase in regulating this system was recognized early, and the potential therapeutic role of phosphodiesterase inhibitors in modulating pathologic conditions was also suggested. At that time, the methylxanthines represented major pharmacologic agents capable of inhibiting cyclic nucleotides and were widely used in respiratory medicine. Initially, bronchodilator effects were considered their major mechanism of action, but subsequent studies suggested other potential roles including an anti-inflammatory one. A number of developments led to the decline in popularity of this class of agents, the foremost being their side-effect profile. The discovery of multiple phosphodiesterase isoforms paired with a better understanding of the physiologic and clinical properties of the phosphodiesterases has re-awakened interest in therapeutic agents in this area and in particular the potential for the development of selective phosphodiesterase inhibitors. Cilomilast is a systemically available, second- generation, selective phosphodiesterase-4 inhibitor. It retains the therapeutic activity of the first generation phosphodiesterase-4 inhibitors (such as rolipram) but is believed to have less of an emetic effect. Cilomilast causes a reduction of tissue cells considered central to the ongoing inflammatory process (macrophages and CD8+ lymphocytes) in patients with chronic obstructive pulmonary disease. Chronic obstructive pulmonary disease is now considered a chronic inflammatory disease of the lungs resulting from prolonged exposure to inflammatory agents in cigarette smoke and other environmental and occupational pollutants, and it is currently the principal target of cilomilast. It is characterized by progressive destruction of parenchymal tissue and punctuated by acute exacerbations. The inflammation is thought to begin in the peripheral airways and lung parenchyma. Chronic obstructive pulmonary disease is a progressive disease, leading to disability and eventual death despite conventional therapy. Cilomilast is completely absorbed following oral administration and has negligible first-pass metabolism. It exhibits low between subject variability. Cilomilast is predominantly protein bound. Plasma clearance is almost entirely metabolic, through multiple parallel pathways. Its terminal elimination half-life is approximately 6.5 hours, and steady state is rapidly achieved. A dose of 15 mg twice daily has been found to be clinically effective. Smoking and age have no clinically relevant effects on cilomilast pharmacokinetics. Most drugs frequently used in patients with chronic obstructive pulmonary disease do not alter its side effect profile. Initial concerns of arteritis involving the gastrointestinal tract in rodent animal models have not been reported in clinical trials. Nausea, presumably of central origin, is the principal adverse reaction seen in healthy subjects taking cilomilast. It has not been associated with the serious cardiac or neurological adverse effects seen with theophylline. Preliminary clinical studies suggest a favorable clinical effect in chronic obstructive pulmonary disease. Cilomilast is generally well tolerated and has not generated safety concerns in reported clinical studies. PMID- 16703121 TI - Varicella-zoster virus vaccine: a review of its use in the prevention of herpes zoster in older adults. AB - Current strategies for managing herpes zoster show variable efficacy and do not prevent its appearance. Varicella-zoster virus vaccine, or "zoster vaccine" is a more potent form of the varicella-zoster virus vaccine currently approved for use in the prevention of varicella in children. Zoster vaccine decreases the incidence of herpes zoster and burden of illness in adults aged 60 years and older and appears more efficacious in patients aged 60-69 than in those over 70 years. Importantly, the incidence of postherpetic neuralgia is significantly reduced in patients who receive zoster vaccine, irrespective of age or sex. The duration of postherpetic neuralgia is also significantly reduced. Zoster vaccine has a favorable safety profile; most treatment-related adverse events are related to the site of injection. This review summarizes the current data on the clinical efficacy and safety of zoster vaccine in adults aged 60 years and older. PMID- 16703122 TI - Ramelteon: profile of a new sleep-promoting medication. AB - To date the mainstay of the pharmacological treatment of insomnia has involved the modulation of the gabaminergic system via benzodiazepines or the Z-drugs, zolpidem, zopiclone or zaleplon. A new approach has explored the melatoninergic system, namely activation of MT1 and MT2 receptors in the suprachiasmatic nucleus of the hypothalamus. Ramelteon (TAK-375) is a novel sleep-promoting agent that acts as an agonist at these receptors; its preclinical pharmacology, mode of action, pharmacokinetics, drug interactions, clinical efficacy, and safety and tolerability are reviewed here. PMID- 16703123 TI - Docetaxel: a tubulin-stabilizing agent approved for the management of several solid tumors. AB - Docetaxel is a semisynthetic taxane that acts by binding to the beta-tubulin subunit of the microtubules, resulting in cell-cycle arrest and apoptosis. It is approved for the management of early and advanced breast cancer, locally advanced and metastatic lung cancer and hormone refractory prostate cancer. Docetaxel has also shown significant antitumor activity in ovarian and gastric tumors and has very recently been approved for the treatment of advanced gastric cancer. Severe neutropenia is the major dose-limiting toxicity with the approved three-weekly regimens, although alternate weekly schedules with less myelotoxicity have been developed for patients with poor bone marrow reserve. This article will review the pharmacology and trials leading to the clinical approval of this agent. PMID- 16703124 TI - New challenges in fullerene chemistry. AB - The aim of this focus article is to emphasize those aspects of fullerene chemistry which currently represent important challenges for the development and future applications of these new carbon allotropes, as well as to celebrate the tenth anniversary of the fullerenes Nobel Prize. PMID- 16703126 TI - Polymer microarrays for cellular adhesion. AB - Microarray screening of polymer libraries for cellular adhesion was developed utilising a thin film of agarose to allow unsurpassed localisation of cell binding onto the array substrate and the discovery of cell specific polymers. PMID- 16703125 TI - Anion-templated assembly of interpenetrated and interlocked structures. AB - The rational development of a general anion templation strategy for the construction of a variety of interpenetrated and interlocked molecular structures based upon the coupling of anion recognition with ion-pairing is described. The success of this anion templation methodology is demonstrated with the halide anion directed assembly of a series of novel [2]pseudorotaxanes containing pyridinium, pyridinium nicotinamide, imidazolium, benzimidazolium and guanidinium threading components and anion binding macrocyclic ligands. Interlocked [2]rotaxane and [2]catenane molecular structures are also synthesised using this anion templation protocol. These interlocked structures feature unique topologically defined hydrogen bond donating binding domains that exhibit a high degree of selectivity for chloride, the templating anion. A series of rhenium(I) bipyridyl containing [2]pseudorotaxane assemblies and a [2]rotaxane further highlight the potential this strategic anion templation approach has in future chemical sensor design and fabrication. PMID- 16703127 TI - Effect of residual monomer on the spectroscopic properties of polythiophenes. AB - The addition of some small molecules can red shift UV-Visible absorption and quench the fluorescence of poly(3-octadecylthiophene). PMID- 16703128 TI - Relative importance of X...O=C vs. X...X halogen bonding as structural determinants in 4-halotriaroylbenzenes. AB - The structures of 4-chloro- and 4-bromotribenzoylbenzene, as well as a solid solution prepared from these two components, are isomorphous and dominated by C X...O=C interactions, whereas type-II I...I interactions are important in the 4 iodo derivative. PMID- 16703129 TI - A novel strategy for fluorescence enhancement in the solid-state: affording rigidity to fluorophores packing. AB - Solid-state fluorescence enhancement was achieved by preparation of rigid packing that was afforded by disposition of benzylamine into tubulate spaces, serving as a powerful and useful strategy for the enhancement. PMID- 16703130 TI - Functionalisation of carbon nanohorns. AB - The functionalisation of single wall carbon nanohorns via 1,3-dipolar cycloaddition has been achieved, and the products have been characterised by spectroscopy, microscopy and thermogravimetry. PMID- 16703131 TI - 2-Amino-3,4-diethylpyrrole derivatives: new building blocks for coiled structures. AB - The synthesis of a multicomponent, mixed oligomer containing alpha-aminopyrrole is described; this system adopts a coiled structure in the solid state and serves as a prototype of a possible new class of hydrogen bond based helicates. PMID- 16703132 TI - Copolymerisation of Pt-carbonyl clusters with Lewis acids: synthesis and crystal structure of the molecular [Cd2Cl4[Pt9(CO)18]2-]infinity 1-D polymer. AB - Reaction of [NBu4]2[Pt9(CO)18] with the soft Lewis acid CdCl2 gives the [Pt9(CO)18(micro3-CdCl2)2]2- adduct, which self-assembles upon crystallization into a 1-D [[Pt9(CO)18(micro3-CdCl2)2]2-]infinity polymer via the formation of chloride bridges. PMID- 16703133 TI - Unusual variations in the incidence of Z' > 1 in oxo-anion structures. AB - A series of Cambridge Structural Database studies show that ionic species generally form low Z' structures, even in those cases where charge assisted hydrogen bonding is a key feature, e.g. oxo-anion complexes. By introducing a competing pi-pi stacking interaction, two oxo-anion compounds are shown to crystallise with more than one molecule in the asymmetric unit, including the first hydrogen phosphate containing structure to have Z' > 2. PMID- 16703134 TI - Supramolecular cruciforms. AB - By self-assembling fluorescent pyridyl cruciforms with bis-Pd-pincer complexes, supramolecular assemblies with attractive optical properties were synthesized; these assemblies show potential as electro-optical materials. PMID- 16703135 TI - An achiral form of the hexameric resorcin[4]arene capsule sustained by hydrogen bonding with alcohols. AB - The well-known hexameric capsules sustained by self-assembly of resorcin[4]arenes 1 with water molecules (1(6).(H2O)8) are shown to assemble similarly with (+/-)-2 ethylhexanol (2EH) as an achiral 1(6).(2EH)6.(H2O)2 species which further encapsulates 2EH. PMID- 16703136 TI - An electrochemical/photochemical information processing system using a monolayer functionalized electrode. AB - An electroactive and photoisomerizable monolayer associated with a Au electrode acts as a Write-Read-Erase information processing system and as a flip-flop Set/Reset memory element. PMID- 16703137 TI - Nanoassembly of biocompatible microcapsules for urease encapsulation and their use as biomimetic reactors. AB - Biocompatible polypeptide capsules with high enzyme loading and activity prepared by templating mesoporous silica spheres were used as biomimetic reactors for performing CaCO3 synthesis exclusively inside the capsule interior via urease catalyzed urea hydrolysis. PMID- 16703138 TI - Non-covalent binding of fullerenes and biomolecules at surface-supported metallosupramolecular receptors. AB - In-situ scanning tunneling microscopy study reveals that two-dimensional metallosupramolecular receptors bind a single or a discrete number of cystine, C60, or diphenylalanine molecules reversibly through non-covalent interactions. PMID- 16703139 TI - Hemiaminals as substrates for sulfur ylides: direct asymmetric syntheses of functionalised pyrrolidines and piperidines. AB - Phenyl stabilised chiral sulfur ylides react with five-membered-ring hemiaminals to give functionalised pyrrolidines directly with high enantioselectivity. The reaction can be diverted to give piperidines instead by isolation of the intermediate epoxide and treatment with TMSOTf. PMID- 16703140 TI - Asymmetric hydrogenation of quinolines catalyzed by iridium complexes of BINOL derived diphosphonites. AB - A chiral diphosphonite, derived from BINOL and with an achiral diphenyl ether backbone, is an excellent ligand for the Ir-catalyzed asymmetric hydrogenation of quinolines; achiral P-ligands serving as possible additives (ee = 73-96%). PMID- 16703141 TI - Sandwich-shaped silver(I) metallomacrocycles encapsulating a XF6(2-) (X = Si, Ge and Sn) anion. AB - A series of sandwich-shaped complexes based on two square tetranuclear Ag(I) metallomacrocycles, [[Ag4(pprd)4]2(XF6)]-(BF4)6.8MeNO2 (pprd = 4-(2 pyridyl)pyrimidine; X = Si, Ge and Sn), in which a XF6(2-) anion is encapsulated, were prepared and their structures were characterized both in the solid state and solution. PMID- 16703142 TI - Bond length and bond multiplicity: sigma-bond prevents short pi-bonds. AB - Analysis of model compounds such as Fe2(CO)6, C2 and HBBH shows that pi-bonds left to themselves are shorter than sigma-bonds; in many ways sigma-bonds prevent pi-bonds from adopting their optimal shorter distances. PMID- 16703143 TI - Sieving behaviour of nanoscopic pores by hydrated ions. AB - In this study, for the first time, the anion dependency of Ag-deposition on self assembled monolayers (SAMs) with alkyl chains long enough to meet the densely packed and well-organized surface is reported. Irrespective of pH, types of terminal groups of the SAMs, and the convective mass transfer condition, SAM structures show the "sieving behaviour" to the Ag deposition by the composition of the electrolytes. PMID- 16703144 TI - Selective functionalization of imidazoles via an iodine-copper exchange reaction. AB - The reaction of protected 4,5-diiodoimidazoles with (PhMe2CCH2)2CuLi regioselectively provides 5-cuprated imidazoles, which readily react with various electrophiles furnishing functionalized imidazoles in good yields; remarkably, these resulting mono-iodoimidazoles undergo again an iodine-copper exchange reaction in the presence of sensitive functional groups, like an aldehyde or a ketone. PMID- 16703146 TI - Convenient, scalable and flexible method for the preparation of imidazolium salts with previously inaccessible substitution patterns. AB - A high yielding and modular approach to N,N'-disubstituted imidazolium salts is described, providing access to substitution patterns that are beyond the reach of established methodology. PMID- 16703145 TI - Highly stable cyclic dimers based on non-covalent interactions. AB - Highly stable cyclic dimers have been assembled through a combination of non covalent interactions, including multiple hydrogen bonding, parallel stacking and hydrophobic shielding. PMID- 16703147 TI - Facile and reproducible syntheses of bis(dialkylselenophosphenyl)-selenides and diselenides: X-ray structures of ((i)Pr2PSe)2Se, ((i)Pr2PSe)2Se2 and (Ph2PSe)2Se. AB - Facile and reproducible methods for the syntheses of bis(di-iso propylselenophosphinyl)selenide ((i)Pr2PSe)2Se (1), bis(di-iso propylselenophosphinyl)diselenide ((i)Pr2PSe)2Se2 (2) and bis(di phenylselenophosphinyl)selenide (Ph2PSe)2Se (3) is reported. PMID- 16703148 TI - Metal complexes of selenophosphinates from reactions with (R2PSe)2Se: [M(R2PSe2)n] (M = Zn(II), Cd(II), Pb(II), In(III), Ga(III), Cu(I), Bi(III), Ni(II); R = (i)Pr, Ph) and [Mo(V)2O2Se2(Se2P(i)Pr2)2]. AB - The reactions of bis(dialkylselenophosphinyl)selenide with a series of metals have been investigated: synthesis of several metal selenophosphinate complexes and their structures are reported. PMID- 16703149 TI - Switching a molecular shuttle on and off: simple, pH-controlled pseudorotaxanes based on cucurbit[7]uril. AB - Cucurbit[7[uril, the wheel component in two new and structurally simple pseudorotaxanes, shuttles between the two axle termini at low pH, whereas the shuttling motion stops at higher pH, as the wheel is bound to the center of the axle. PMID- 16703150 TI - Synthesis of crystallized mesoporous transition metal oxides by silicone treatment of the oxide precursor. AB - Ordered mesoporous transition metal oxides were successfully crystallized after strengthening the amorphous framework by a silica layer, which efficiently protected the original mesoporous structure against crystallization and resulting mass transfer. PMID- 16703151 TI - Electrochemical STM observation of new structures of CO adsorbed on a Pt(111) electrode surface. AB - Presented are two newly observed adstructures of adsorbed CO onto Pt(111), (2 x 2)-3CO-beta and (2 x 2)-4CO, observed during the structural evolution from the well-known (2 x 2)-3CO-alpha structure to the (square root 19 x square root 19) 13CO structure. PMID- 16703152 TI - Writing for scientific publication: tips for getting started. AB - Pediatricians in training and clinical practice may be eager to submit their first scientific publication by writing a letter to the editor, case report, review article, or short clinical series. Unfortunately, the would-be author encounters a literary developmental delay when facing the blank page or computer screen. This article is dedicated to the novice author and offers practical tips on writing the first manuscript. Authors are encouraged to draw from their extensive experience writing other materials and to approach the task in a systematic way. Computer technology and software developments should facilitate the process. PMID- 16703153 TI - Turner syndrome: an update and review for the primary pediatrician. AB - Turner syndrome (TS) is among the most common of the sex chromosomal aneuploidies. It results from the absence of one sex chromosome (or part of an X chromosome) in a female, leaving only one X chromosome present in the cell. Primary care physicians should be able to recognize the presenting signs and symptoms of TS, and once the diagnosis is confirmed by a chromosome analysis, they should be able to serve as a valuable source of support for the patient and her family and understand the most current treatments available. PMID- 16703154 TI - Concordance between Neurologic Screening Test and Neurologic Examination in newborns. AB - The aims of this study were to calculate concordance among results of the assessment of a population of neonates by using a screening version of neurologic assessment developed by our group with the Amiel-Tison examination, to evaluate distribution of the findings for each item in the studied population, and to develop an optimality score to aid in the detection of possible deviations in infants. Infants in this report include 107 infants born consecutively; they were mainly at-term infants, or preterm infants who, when reaching at-term corrected age, were suspected to be abnormal with a wide spectrum of complications. Newborns were tested by neurologic screening and neurologic examination. Agreement, correlations among tests, sensitivity, and specificity were calculated. Correlation among results from Neurological Screening and Neurological Examination was significant (rho = 0.759, p = 0.01), and Cronbach's alpha concordance had an acceptable value (0.83). Sensitivity and specificity of neurologic screening after Amiel-Tison examination comparison were 0.98 and 0.82, respectively. Our results support the use of the neurologic screening proposed. PMID- 16703156 TI - Inner-city adults' knowledge about the effects of cigarette smoking on child health. AB - We sought to determine what adults in an inner-city setting know about the specific effects of adult cigarette smoking on child health. A cross-sectional survey was conducted at an inner-city community health center in the Bronx, New York; 684 subjects participated. Overall, 21% were current smokers, 19% had quit, and 60% had never smoked. While the majority of subjects knew about the effects of smoking on adult health, they were unaware of the extent to which smoking was harmful to child health. Notably, 72% did not know that cigarette smoking increased the risk for ear infections in children, 68% did not know that smoking increased the risk of colds in children, and 61% did not know that smoking increased the risk of sudden infant death syndrome. The findings suggest a need for public health education about the effects of adult smoking on child health. PMID- 16703155 TI - What providers from general emergency departments say about implementing a pediatric asthma pathway. AB - The goal of this study was to assess institutional climate and providers' readiness to adopt pediatric pathways for asthma treatment and management. Twelve focus groups were held with 24 physicians/physicians' assistants, 20 nurses, and 17 emergency medical technicians from emergency departments in 4 general hospitals from July to October 2002. Positive experience with previous pathways, open communication and buy-in from clinicians and administrators, comprehensive training on pathways, and adapting standards to fit specific emergency department environments were identified as necessary elements for pathway adoption. Providers were optimistic about successfully implementing an asthma pathway (95%) and supportive of pathway implementation (87%). PMID- 16703157 TI - Microbial colonization of the hands of residents. AB - To determine whether carriage of resistant Gram-positive organisms by residents increases over time, the dominant hand of pediatric residents was cultured. Among first-year residents, 53 Gram-positive organisms were isolated; 12 were resistant to oxacillin, 13 to clindamycin. Six residents had organisms resistant to both; 14 carried at least one resistant to either. Among third-year residents, 46 Gram positive organisms were cultured; 9 were resistant to oxacillin, 6 to clindamycin. Three residents carried organisms resistant to both; 10 carried at least one resistant to either. Colonization with resistant Gram-positive organisms did not increase with length of training. PMID- 16703158 TI - Evaluation of prolonged aPTT values in the pediatric population. AB - To assess the likelihood of significant bleeding disorders in children with prolonged activated partial thromboplastin times (aPTTs), a retrospective chart review was performed. Data analyses determined that in the absence of symptoms and a negative family history, the diagnosis of a bleeding disorder was unlikely in an individual with a prolonged aPTT (negative predictive value = 80%). Conversely, a prolonged aPTT was predictive (positive predictive value = 62%) in the presence of both clinical symptoms and a documented family history. The scope of laboratory investigation in any child with a prolonged aPTT should be tempered by the clinical presentation and the associated personal and family histories. PMID- 16703160 TI - Narcolepsy with cataplexy in early childhood. AB - Narcolepsy is a rare neurologic sleep disorder with morbidity associated with functional impairment and frequent delay in diagnosis. Symptoms typically manifest in adolescence or early adulthood, but diagnosis of narcolepsy has been reported in early childhood. Diagnosis rates are as low as 50% of the total population of patients with narcolepsy and are delayed as much as 10 years after disease onset due to inadequate patient-physician communication and/or misdiagnosis. I present the complexity of diagnosing narcolepsy in early childhood in a patient with cataplexy that started soon after independent ambulation at age 10 months. PMID- 16703159 TI - Parent reasons for nonreturn to a pediatric weight management program. AB - To identify parent reasons for lack of return to a weight management program, a questionnaire was sent to 85 families who attended two or fewer visits; 43 families completed the questionnaire. A total of 37% reported that the program was not what they were looking for. Specifically, they were dissatisfied with the staff or treatment focus. Other reasons for nonreturn were distance to program (23%), scheduling conflicts (21%), and lack of insurance for weight management (21%). High body mass index z score was associated with return, but age, gender, race, and medical condition were not. Matching program structure, location, and time to parent preference might improve retention. PMID- 16703161 TI - Congenital metopic craniosynostosis with secondary trigonocephaly: a case report. PMID- 16703162 TI - Congenital livedo reticularis and recurrent strokes in two unrelated young children. PMID- 16703163 TI - A recurrent presentation of hand, foot, and mouth disease. PMID- 16703164 TI - Cough in a teenager. PMID- 16703165 TI - Cough in a teenager. Commentary. PMID- 16703166 TI - Paresis. PMID- 16703168 TI - Picky eater. PMID- 16703167 TI - Paresis. Commentary. PMID- 16703170 TI - The appearance, nature, and significance of cytoplasmic accumulations, as exemplified by the granular cell change. PMID- 16703171 TI - Evaluation of RB gene and cyclin-dependent kinase inhibitors P21 and P27 in pleomorphic xantoastrocytoma. AB - Pleomorphic xantoastrocytoma (PXA) is a rare, circumscribed astrocytic tumor that usually occurs in the superficial cerebral hemispheres in children and young adults. Most patients have a favorable prognosis, but recurrence and malignant transformation have been reported. In diffuse gliomas, approximately one third demonstrate mutations of the RB gene. Low expression level and high activity of p27 are known to constitute an independent prognostic factor in patients with malignant gliomas, while p21 expressions have variable labeling ranges. The molecular and genetic basis for tumorigenesis and progression of PXA are still largely unknown. In this study, 13 PXAs were examined immunohistochemically for pRb, p21, and p27 expression. Nine PXAs expressed homogeneous pRb positivity in the most nuclei of the tumor cells. Four cases showed an abnormal pRb staining pattern. All PXAs were positive for nuclear expression of p21. Diffuse nuclear positivity of p27 was seen in 10 cases, focal in 2, and in 1 case was not present. The cases with focal and negative p27 nuclear expression had few pRb positive nuclei. The majority of PXAs appear to have preserved pRb, p21, and p27 functions. Additional studies are necessary to investigate whether cases with altered pRb and p27 expressions are associated with increased risk of recurrence or malignant transformation. PMID- 16703172 TI - Chondromatous hamartoma of the chest wall: description of 3 new cases and literature review. AB - Chondromatous hamartoma of the chest wall is an extremely rare, benign lesion that usually occurs in early infancy. It typically arises in the rib cage and produces a large mass. It is composed of a varying admixture of hyaline cartilage that has features resembling growth plate cartilage, fascicles of spindle cells, woven bone, and hemorrhagic cysts. Treatment consists mainly of surgical excision, which is usually curative. We present 3 new cases, which demonstrated interesting findings, including multicentricity, involvement of the sternum, and local recurrence, and we discuss these findings in the context of a literature review. PMID- 16703173 TI - CD10, actin, and vimentin expression in breast phyllodes tumors correlates with tumor grades of the WHO grading system. AB - The discrimination of borderline from malignant primary breast phyllodes (PT) tumor is still unclear. We studied 22 PT cases to investigate the immunohistochemical expression (staining of stromal CD10, SMA [smooth muscle actin], and vimentin) as well as the features of focal glandular atypia to determine whether these correlated with the histopathologic grading system. In our results, the stromal staining of CD10 was positive in 4 of 6 malignant and 2 of 5 borderline PT cases, but negative in all benign PT cases. Stromal actin and intraglandular vimentin-expressive tumor cells were found in 5 of 6 malignant PT cases but not in borderline and benign PT cases. There is a significant difference in the panel of stromal CD10, actin, and vimentin expression between borderline and malignant PT (p<0.05). Besides, the progression of malignant potential breast phyllodes tumor may cause glandular epithelium atypia with loss of polarity. PMID- 16703174 TI - Colonic and duodenal flat adenomas in children with classical familial adenomatous polyposis. AB - Flat adenomas of the colon and duodenum have been described as associating with familial adenomatous polyposis (FAP), its attenuated variant, and the so-called hereditary nonpolyposis colorectal cancer. There seem to be no report on the occurrence of flat adenomas in pediatric patients with family history of FAP. We are reporting 4 children from 2 cancer-prone families in whom colonic and duodenal moderately dysplastic flat adenomas were found. Gastrointestinal endoscopy and biopsies were performed in 3 female siblings (7, 9, and 11 years old) and 1 male (9 years old) when referred for screening owing to familial history of bowel cancer (family 1) or evidence of bilateral congenital hypertrophy of the retinal pigment epithelium (CHRPE), which is known to be associated with FAP (family 2). Endoscopic visualization of the mucosa was improved by use of 0.2% indigo carmine solution spray. Biopsies were routinely processed for H&E and immunohistochemistry staining. Present patients were asymptomatic, with the exception of 2 weeks rectal bleeding in 1 of them. The colonic videoendoscopy showed in 2/3 siblings hundreds of flat or slightly raised plaques less than 1 cm in diameter as well as some classic polyps throughout the colon. The other sibling showed 40 flat-topped lesions with minimal elevation and central umbilication in the cecum. Upper endoscopy demonstrated a few flat lesions in the nonperiampullary area of the duodenum in 2/4 patients. The colonic videoendoscopy performed on the 9-year-old boy revealed multiple small sessile polyps. Microscopic study demonstrated tubular adenomas with a few neoplastic crypts, slight disarray of the overall architecture, and moderate (low-grade) dysplasia of the epithelium. These features were more obvious at the center and superficial areas of the adenomas. The 4 children had multiple flat adenomas of the colon and duodenum (2/4) matching with those described in adult patients. Flat adenomas in the context of FAP probably represent early stages of the adenoma development. Careful endoscopic-histologic correlation may result in increasing recognition of these lesions at the pediatric age. PMID- 16703175 TI - Images in pathology. Russell body gastritis in an HIV-positive patient. PMID- 16703176 TI - Images in pathology. Tumor spreading along an incisional scar line. PMID- 16703177 TI - Images in pathology. Dove in flight: who will save the dove? PMID- 16703178 TI - Images in pathology. Leiomyosarcoma twins. PMID- 16703179 TI - Images in pathology. Florid (floral) synovitis. PMID- 16703180 TI - Well-differentiated chondrosarcoma of the humerus with prominent granular cell component: a hitherto unreported occurrence. AB - We report the case of a well-differentiated chondrosarcoma of the proximal humerus in a 60-year-old man that featured a prominent component of granular cells, the granules being shown by electron microscopy to be lysosomal. Although secondary granular cell changes have been described in a large variety of tumor types, this is, to the best of our knowledge, the first documentation of this phenomenon in a skeletal chondrosarcoma. PMID- 16703181 TI - Early post-transplant smooth muscle neoplasia of the colon presenting as diminutive polyps: a case complicating post-transplant lymphoproliferative disorder. AB - A 44-year-old woman, 3 years post-transplant for pulmonary sarcoidosis, developed abdominal pain and diarrhea 13 months subsequent to an eradicated diffuse large B cell-type, post-transplant lymphoproliferative disorder (PTLD) of the cecal region. Endoscopic examination identified multiple pale tan 5-to-9 mm rubbery nodules of the transverse and right colon in an otherwise unremarkable mucosa. Histology was characterized by bland smooth muscle proliferations, focally pushing into the mucosa. Immunohistochemistry (IHC), in situ hybridization (ISH), and polymerase chain reaction (PCR) of the sampled nodules confirmed Epstein-Barr virus (EBV) infection of neoplastic cells. To our knowledge, this is the first reported case of EBV-related post-transplant lymphoproliferative and smooth muscle neoplasms (PTSN) having distinct tropism for the colon. Endoscopic features of early PTSN, which in this case presented as diminutive polypoid lesions, have not been described previously. PMID- 16703182 TI - Extranodal marginal zone B-cell lymphoma/maltoma of the lip in a child: case report and review of cutaneous lymphoid proliferations in childhood. AB - All forms of cutaneous lymphomas are rare in children. Extranodal marginal zone B cell lymphomas (EMZBL)/mucosa-associated lymphoid tissue (MALT) lymphomas are unusual neoplasms in children and young adults. We report a case of an EMZBL/MALT lymphoma of the lip in a previously healthy 14-year-old boy without immunodeficiency, confirmed by immunohistochemistry and documentation of clonal rearrangement of the immunoglobulin heavy-chain gene. Additionally, we present a review of the differential diagnosis of skin and mucosal lymphoid proliferations in childhood. PMID- 16703183 TI - Carcinoma of the neck showing thymic-like elements (CASTLE): report of a case and review of the literature. AB - Carcinoma showing thymic-like elements (CASTLE) is a rare tumor affecting thyroid and neck soft tissues, which has to be distinguished from squamous cell and anaplastic thyroid carcinoma, because it has a better prognosis. We report a new case of CASTLE which occurred in a patient submitted to total thyroidectomy with central neck dissection. The tumor stained positively for CD5, which seems to be the most useful marker in the differential diagnosis. By the analysis of the 18 cases reported in literature, total thyroidectomy with selective modified neck dissection should be the treatment of choice and radiotherapy should be considered for patients with positive nodal status. PMID- 16703184 TI - Skin adnexal neoplasm closely resembling adenomatoid tumor: a unique occurrence. AB - We describe a primary skin neoplasm located in the left chest wall that closely resembled adenomatoid tumor of male and female genital tract. It occurred in a 52 year-old woman who had undergone a left quadrantectomy with regional lymphadenectomy for invasive ductal carcinoma of the breast 7 years previously. The tumor involved the dermis and subcutaneous tissue and measured 0.7 cm in greatest diameter. Immunohistochemical evaluation showed strong positivity for cytokeratin pool and negativity for CD31, calretinin, WT-180, and for estrogen and progesterone receptors. We are unaware of a previous description of this morphologic pattern in a primary skin tumor, which we have interpreted as of skin adnexal and specifically of eccrine sweat gland origin. PMID- 16703185 TI - Prevention of positive coronary artery remodeling with statin therapy in patients with coronary artery diseases. AB - Since positive coronary artery remodeling with large plaque burden is associated with subsequent coronary events, the authors tested their hypothesis that secondary prevention of coronary events by a statin may be associated with inhibition of the process of positive coronary artery remodeling in underlying coronary atherosclerotic lesions in patients with coronary artery diseases. They evaluated the intravascular ultrasound imaging in angiographically normal coronary lesions at baseline and after 6 months of therapy in 64 patients with coronary artery diseases. External elastic membrane area was defined as the vessel area, and the difference between the vessel and lumen area was calculated as plaque area. The relative echogenicity of coronary plaque to adventitia was evaluated as acoustic characteristics of coronary plaque. Twenty-five patients were treated with a statin and 39 patients did not receive a statin. In patients treated with a statin, plaque area decreased by 12% (p = 0.013) compared to an increase in plaque area of 13% (p = 0.023) in those who did not receive a statin. The vessel area was not enlarged in patients treated with a statin but did show positive remodeling in patients who had plaque progression without a statin. The relative echogenicity of plaque was unchanged in patients treated with a statin but significantly decreased in patients not receiving a statin. A statin may prevent positive coronary artery remodeling via inhibition of plaque progression in underlying coronary atherosclerotic lesions in patients with coronary artery diseases. PMID- 16703186 TI - Increased serum leptin concentrations in patients with chronic stable angina pectoris and ST-elevated myocardial infarction. AB - Leptin is an adipocytokine that is produced mainly by adipose tissue; it is also identified in atherosclerotic lesions in human coronary atherosclerosis. However, the relation of serum leptin concentrations to ischemic heart disease (IHD) is still obscure. The aims of the present study were to investigate serum leptin concentrations in patients with ST-elevated myocardial infarction (STEMI) and with chronic stable angina pectoris (CSAP) and to evaluate the possible correlations of leptin to other atherosclerotic risk factors; including serum high sensitive C-reactive protein (Hs-CRP), serum homocysteine, and fibrinogen concentrations. For this purpose, 35 patients with CSAP, 40 with acute STEMI, and 30 control subjects with normal findings from coronary angiography were taken into the study prospectively. Serum leptin concentrations were significantly higher in patients with CSAP and STEMI compared to the control group (7.74 +/ 1.34 vs 6.37 +/-1.85 ng/mL, p=0.021 and 8.22 +/-3.13 vs 6.37 +/-1.85 ng/mL, p=0.023, respectively). In addition, serum homocysteine concentrations were significantly increased in patients with CSAP (15.23 +/-5.96 vs 11.40 +/-2.11 micromol/L, p=0.025) and patients with STEMI (15.90 +/-5.02 vs 11.40 +/-2.11 micromol/L, p=0.012) compared to the control group. Serum fibrinogen concentrations were significantly increased only in the CSAP group as compared to controls (4.15 +/-1.39 vs 3.45 +/-1.19 g/L, p=0.025). No significant correlation was found between leptin levels and selected risk factors. In conclusion, serum leptin concentrations were significantly higher in both the CSAP and STEMI groups. However, owing to the lack of correlation between the leptin levels and selected classical coronary risk factors, it may be considered that leptin can be evaluated as one of the independent risk factors for IHD. Further randomized and controlled studies will be required to determine the pathophysiological meaning of the increased leptin levels and the central role between adipocyte function and atherosclerosis. PMID- 16703187 TI - Impacts of glycoprotein IIb/IIIa inhibition on QT dispersion after successful percutaneous coronary intervention. AB - Coronary ischemia augments inhomogeneity in ventricular repolarization. Decrease in the QT dispersion (QTd) following restoration of coronary blood flow to the ischemic myocardium by successful percutaneous coronary intervention (PCI) is an expected outcome. The purpose of the study was to seek whether glycoprotein IIb/IIIa (GP IIb/IIIa) inhibition has additional beneficial effects on QT dispersion after angiographically successful PCI. The study involved 111 consecutive patients scheduled for elective coronary balloon angioplasty with or without stent implantation. Sixty patients (mean age 58 +/-9) were randomized to receive standard therapy including preprocedural aspirin, ticlopidine, and IV heparin, and 51 patients (mean age 54 +/-10) were randomized to receive additional IV tirofiban infusion before the lesion was crossed with the guidewire. Standard 12-lead simultaneous ECG recordings for the measurement of QTd and corrected QTd (QTcd) (calculated by using Bazett's formula) were obtained before and immediately after the procedure, and at the 6th, and 24th hours. Blood samples for detection of postprocedural myocardial damage (CK-MB and cTn-I) were taken before and immediately after the procedure, at the 6th, 12th, and 24th hours. In total, 128 stenoses were treated with PCI. Seventy of these lesions were in the standard therapy group and 58 in the tirofiban group. QTd and QTcd were not statistically different between the 2 groups before and immediately after the procedure and at the 6th hours, but at the 24th hour QTd and QTcd were significantly longer in the standard therapy group (p=0.047 and p=0.001, respectively). Postprocedural troponin-I elevation (B=0.692, p=0.037), maximum inflation pressure (B=0.182, p=0.001), and previous myocardial infarction (MI) (B=0.885, p=0.004) were defined as the predictors of the final QT dispersion at the 24th hour. QT dispersion significantly decreased after successful percutaneous coronary intervention. GP IIb/IIIa inhibition therapy was not superior by means of recovery of increased QT dispersion during the early hours of the intervention, but it prevented minor myocardial necrosis and provided more long-lasting recovery in QT dispersion as compared with heparin therapy. This impact of GP IIb/IIIa receptor inhibition on QTd may be a possible mechanism by which these drugs reduce cardiovascular events after PCI. PMID- 16703188 TI - In-hospital and long-term prognostic value of fibrinogen, CRP, and IL-6 levels in patients with acute myocardial infarction treated with thrombolysis. AB - Plasma fibrinogen, C-reactive protein (CRP), and interleukin-6 (IL-6) levels in patients with acute myocardial infarction (AMI) receiving thrombolysis have been related to prognosis. The aim of this study was to investigate the time course of plasma fibrinogen, CRP, and IL-6 levels during the in-hospital phase in patients with AMI receiving thrombolysis, and their relationship to in-hospital and prognosis after 12-months follow-up. In 40 patients presenting with AMI within 6 hours of symptom onset and treated with thrombolysis, plasma fibrinogen, CRP, and IL-6 levels were measured on admission and after 6, 12, 24, 48, and 72 hours; 7 days; and 6 months. Patients with other diseases that can alter fibrinogen, CRP, or IL-6 levels were excluded. Patients had a clinical follow-up at 6 and 12 months, and the following cardiac events were recorded: cardiac death, recurrent angina, recurrent AMI, and heart failure. Plasma fibrinogen concentrations decreased significantly (p <0.01 vs admission levels) at 12 hours (425 +/-94 vs 322 +/-132 mg/dL), started to increase at 24 hours, reached peak value at 72 hours (602 +/-209 mg/dL), remained elevated at 7 days, and were back to admission levels at 6 months (375 +/-79 mg/dL). CRP levels increased significantly at 12 hours (0.73 +/-0.43 vs 0.23 +/-0.11 mg/dL, p <0.01), reached peak value at 72 hours (7.66 +/-3.28 mg/dL), decreased significantly on day 7 (2.32 +/-1.17 mg/dL), and at 6 months were within normal limits (0.49 +/-0.29 mg/dL). IL-6 levels increased significantly at 6 hours (14.03 +/-8.13 vs 6.37 +/-3.88 pg/mL, p <0.05), reached peak value at 24 hours (59.49 +/-23.57 pg/mL), started to decrease at 48 hours, and at 6 months were within normal limits (2.25 +/-1.24 pg/mL). During the in-hospital phase 33 patients had an uneventful course and 7 patients had complications (3 post-AMI angina; 4 heart failure). During the 12 month follow-up period 28 patients had an uneventful course, and 12 patients had complications (1 cardiac death, 5 recurrent angina, 2 recurrent AMI, and 4 heart failure). Regarding the in-hospital prognosis, fibrinogen, CRP, and IL-6 levels were significantly higher (p <0.05) in patients with complications from 48 to 72 hours, from 12 hours until day 7, and from 6 hours until day 7, respectively. During the 12-month follow-up period fibrinogen, CRP, and IL-6 levels were significantly higher in patients with complications (at 48, 24, and 24 hours, respectively) only in the subgroup of patients who had complications within the first 6 months following AMI. Multivariate analysis showed that CRP at 48 hours was the most important factor related to in-hospital prognosis (p = 0.02), and ejection fraction followed by CRP at 24 hours (p = 0.02) to 6-month prognosis (p = 0.018). Fibrinogen, CRP, and IL-6 levels alter in patients with AMI receiving thrombolysis, and are related both to in-hospital and to 6-month follow-up prognosis. PMID- 16703189 TI - High-sensitivity C-reactive protein in patients with metabolic syndrome. AB - High-sensitivity C-reactive protein (CRP) has been shown to predict cardiovascular disease. Metabolic syndrome has been found to play a critical role in the development of cardiovascular disease. The purpose of this report is to assess the relationship between CRP and the metabolic syndrome. A total of 50 patients with metabolic syndrome and 40 healthy persons were included in the study. Plasma concentrations of CRP were measured by means of particle-enhanced immunonephelometry with the Behring nephelometer using N Latex CRP mono reagent. CRP levels were higher in patients with metabolic syndrome than control group (10.6 +/-5.4 mg/L vs 3.5 +/-0.8 mg/L, p<0.001). In partial correlation, plasma CRP positively correlated with body mass index (p<0.001), waist circumference (p<0.001), waist-to-hip ratio (p<0.01), total cholesterol (p<0.001), LDL cholesterol (p=0.033), triglyceride (p=0.023), and fasting blood glucose (p=0.043) in patients with metabolic syndrome. HDL-cholesterol did not significantly correlate with CRP (p>0.05). In multiple regression analysis, body mass index (p<0.01), waist circumference (p<0.01), and fasting blood glucose (p<0.01) showed independent correlations with plasma CRP. CRP levels were found higher in patients with metabolic syndrome. These results suggest that abdominal obesity is the critical correlates of elevated plasma CRP levels found in patients with metabolic syndrome. These patients carrying high risk for cardiovascular events must be followed closely. PMID- 16703190 TI - Total serum insulin-like growth factor-1 and C-reactive protein in metabolic syndrome with or without diabetes. AB - There are only a few data on the relationship of insulin-like growth factor-1 (IGF-1), implicated in glucose homeostasis, and C-reactive protein (CRP), a measure of subclinical systemic inflammation, in patients with the metabolic syndrome (MetS). The authors investigated, in a cross-sectional design, the correlation between total IGF-1 and CRP in 170 subjects. Among them 123 had the MetS (National Cholesterol Program ATP III definition) and 47 did not, and 136 had type 2 diabetes mellitus (DM) and 34 did not. Anthropometric variables, clinical characteristics, as well as laboratory measurements, including total IGF 1 and CRP, were recorded. CRP levels showed a significant negative correlation with total IGF-1 concentrations, both in the whole study population (r = -0.252, p = 0.001) and the MetS group (r = -0.203, p = 0.025), regardless of the presence of DM. This correlation remained significant after adjusting for age, gender, smoking status, and waist circumference (r = -0.18, p = 0.05). Both low IGF-1 and high CRP levels had an almost linear relationship with the number of MetS components (p = 0.029 and p = 0.020, respectively), suggesting a close relationship of both variables with the cardiovascular disease (CVD) risk involved. The correlation between high CRP and low total IGF-1 might indicate that an increase in CRP levels may well be a key factor for the reduction in IGF 1 concentrations. Both factors are related to an increase in risk for MetS and CVD and this finding might have clinical implications in preventing or treating MetS, DM, and CVD. Given the cross-sectional design of the study, this finding should be confirmed by larger prospective and, it is hoped, interventional studies. PMID- 16703192 TI - Venous hypertension induces increased platelet reactivity and accumulation in patients with chronic venous insufficiency. AB - The objective of this study was to determine whether there are changes in platelet activation and rheology in patients with chronic venous insufficiency (CVI) and what their impact is on this disease. Anticoagulated peripheral venous blood collected from 21 patients with CVI and 13 normal control subjects in different bodily positions was incubated either with 0.5 mumol/L adenosine diphosphate (ADP) or without agonist and analyzed by whole blood flow cytometry. Soluble P-selectin was analyzed in obtained sera by enzyme-linked immunosorbent assay. Platelet count was determined by a whole blood analyzer. Circulating platelets were more reactive to stimulation with 0.5 mumol/L ADP in patients with CVI compared with control subjects. There was no statistically significant change in platelet activation without ADP and the level of soluble P-selectin as a function of posture. Under simulated venous hypertension, platelet accumulation was observed in patients with CVI. Patients with CVI had increased platelet reactivity and accumulation during orthostasis, suggesting this might be a contributory factor to CVI pathogenesis. PMID- 16703191 TI - Microalbuminuria is determined by systolic and pulse pressure over a 12-year period and related to peripheral artery disease in normotensive and hypertensive subjects: the Three Areas Study in Greece (TAS-GR). AB - Microalbuminuria and peripheral artery disease represent 2 different forms of target organ damage due to raised blood pressure. The aim of this study was to investigate the association between blood pressure with microalbuminuria and the appearance of peripheral artery disease after more than a decade, and moreover, to address whether any relationship exists between microalbuminuria and peripheral disease in a Greek Caucasian population. In 1990, 635 normal subjects were examined and their blood pressure was recorded. Nine and 12 years later, subjects were reexamined and 361 of them (57%) were available at last visit for the determination of microalbuminuria and ankle-arm index. Microalbuminuria was detected in 35/361 (9.7%) and peripheral artery disease in 89/361 (24.7%). Both conditions were statistically correlated with pulse and systolic blood pressure at all time points during the 12-year follow-up period, while the relationship with diastolic and mean arterial pressure existed only for baseline values and was then abolished. Microalbuminuria was statistically correlated to peripheral artery disease (r = -0.460, p = 0.0001). Blood pressure levels seemed to predict the appearance of microalbuminuria and peripheral disease after 12 years. Microvasculature and macrovasculature abnormalities (microalbuminuria and peripheral disease, respectively) showed a significant relationship, suggesting a common pathogenetic mechanism. PMID- 16703193 TI - Cramps and muscular pain: prevention with pycnogenol in normal subjects, venous patients, athletes, claudicants and in diabetic microangiopathy. AB - The aim of this study was to assess the preventive action of Pycnogenol (Horphag Research Ltd, UK) on cramps and muscular pain in different groups of subjects and patients. The study included a 5-week observation period (4 weeks treatment and one follow-up week after the suspension of treatment) to evaluate the efficacy of Pycnogenol after its withdrawal. Four 50 mg capsules (total dose 200 mg/day) were prescribed with suggestion to drink at least 1.5 liters of water every day. In the first part of the study 66 healthy subjects completed a 5-week follow-up period. The difference between number of cramps attacks recorded within the 2 weeks before inclusion and the number of episodes during the fourth (p <0.05) and fifth (p <0.05) week were statistically significant. In normal subjects the average number of episodes was reduced from 4.8 (1.2) events per week to 1.3 (1.1) at 4 weeks (p <0.05). In venous patients the decrease in events was from 6.3 (1.1) to 2.6 (0.4) per week (p <0.05). In athletes the number of episodes decreased from 8.6 (2) to 2.4 (0.5) (p <0.05). The decrease was still present at 5 weeks in the 3 groups, to levels significantly lower than inclusion values (p <0.05). In the second part of the study, patients with intermittent claudication and diabetic microangiopathy were evaluated and treated (4 weeks). The groups treated with Pycnogenol and the control, placebo groups were comparable. There was a significant decrease in the number of cramps episodes (p <0.05) and in the score concerning muscular pain (p <0.05) in claudicants and diabetics. No significant effects were observed in the placebo groups. In conclusion, cramps and muscular pain, common in these 2 types of patients, were decreased by the use of Pycnogenol. Globally, these results suggest that the use of Pycnogenol prevents cramps, muscular pain at rest, and pain after/during exercise in normals, in athletes prone to cramps, in patients with venous disease, in claudicants, and in diabetics with microangiopathy. The difference is statistically significant considering objective observations (cramps episodes) and evaluating more subjective aspects (score). This indicates that Pycnogenol is effective in reducing pain and cramps during retraining and rehabilitation increasing its efficiency. In starting any physical rehabilitation program, particularly in vascular subjects, the limitation in mobility associated with muscular pain and with cramps tends to be relevant, and controlling these symptoms is useful to speed up the retraining process. PMID- 16703194 TI - Right ventricular volume and mass determined by cine magnetic resonance imaging in HIV patients with possible right ventricular dysfunction. AB - Impaired right ventricular (RV) function has been reported to occur in patients with HIV when studied by echocardiography. However, for accurate evaluation of RV function and morphology, first-pass radionuclide ventriculography (RNV) and cine magnetic resonance imaging (cine-MRI) are methods of choice. Studies of RV involvement in patients with HIV are of interest since pulmonary hypertension is a known serious complication of HIV recognized with increasing frequency. The aim of the present study was to characterize cardiac function and geometry in patients with HIV and reduced right ventricular ejection fraction (RVEF). To do so, we screened patients with RNV and performed an additional cine-MRI in those with reduced RVEF determined by RNV. Ninety patients with HIV were included. To evaluate the MRI measures exactly we included 18 age- and gender-matched healthy volunteers to establish reference values. RNV showed in 13 of the 90 patients a reduced RVEF with a standard cutoff value for RVEF of 0.50. Six of these agreed to have an additional MRI investigation performed. These 6 patients with HIV had an RVEF measured by RNV between 0.41-0.49. Measured by MRI the range of RVEF was 0.47-0.55 with 3 below the lower 95% reference limit according to the control group (lower reference limit: 0.49). None of the 6 patients with HIV had dilated right ventricle and only 1 had a marginally increased right ventricular mass index of 43 g/m(2) (reference: <41 g/m(2)). With use of MRI, a few patients with HIV may have a marginally reduced RVEF but normal RV dimensions and mass. Thus, RV dysfunction does not seem to constitute a major clinical problem in this antivirally treated HIV population. PMID- 16703195 TI - Transcatheter retrieval of different types of central venous catheter fragment: experience in 13 cases. AB - In total, 13 patients, from April 1992 to November 2002 at this institution, had embolization caused by central venous catheter fragments including 4 peripherally inserted central catheters (PICCs), 1 Hickman catheter, 1 Swan-Ganz catheter, and 7 port-A catheters. These dislodged fragments were percutaneously retrieved successfully in 12 patients. As each type of the central venous catheter has its distinct properties, the locations of embolization, detection, and retrieval methods differed. PMID- 16703196 TI - Foot gangrene in patients with end-stage renal disease: a case control study. AB - The prevalence of peripheral arterial disease (PAD) in patients with end-stage renal disease (ESRD) is high, with an annual risk of amputation estimated at 13%, and indications for limb revascularization in patients combining ESRD with stage IV PAD (foot gangrene) are still controversial. This case-controlled study compared survival, limb salvage, and quality of life in a group of patients hospitalized for foot gangrene according to their renal status (ESRD versus no renal insufficiency). All patients with ESRD hospitalized for foot gangrene (n = 16) from 1996 to 2002 were compared with a control group with normal creatininemia (n = 24) hospitalized for foot gangrene due to peripheral atherosclerotic arterial disease. The 2 groups were matched for age, sex ratio, and number with diabetes mellitus. After a mean follow-up of 467 +/-410 days, patients with ESRD had a more severe prognosis as regards mortality (68.7% vs 12.5%, p = 0.0005) and major amputation (31% versus 8%, p = 0.09). The ESRD group was characterized by more frequent extensive arterial calcifications (16/16 vs 13/24, p = 0.002), owing to a higher level of the calcium phosphorus product (3.54 +/-1.2 vs 2.4 +/-0.6, p = 0.0023), and by impaired microcirculatory perfusion, as indicated by a lower oxygen pressure (TcPO(2)) (15.6 +/-12 mm Hg vs 26 +/-16, p = 0.07). ESRD implies a poor prognosis in patients with stage IV peripheral arterial disease. PMID- 16703197 TI - Aortic pseudoaneurysm as a rare cause of superior vena cava syndrome--a case report. AB - This case is about an aortic pseudoaneurysm after Dacron graft replacement that presented as a rapidly evolving superior vena cava syndrome. PMID- 16703198 TI - Gemcitabine and acute myocardial infarction--a case report. AB - The objective of this paper is to report a case of acute, non-q-wave myocardial infarction, presumably secondary to gemcitabine chemotherapy for nonsmall cell lung cancer. A 43-year-old woman with postpartum cardiomyopathy and ischemic heart disease was treated with gemcitabine for metastatic nonsmall cell lung cancer. Three days after her 5th treatment with gemcitabine, she developed chest pain and was diagnosed as having acute, non-q-wave myocardial infarction. She made an uneventful recovery. An objective causality assessment revealed that the adverse event was possible. Gemcitabine has been previously reported to be causative of acute myocardial infarction. Ischemic complications of chemotherapeutic agents are discussed. A review of literature on this subject is presented. Gemcitabine should be administered with caution in patients with underlying cardiac disease. PMID- 16703199 TI - Incomplete ventricular septal rupture following blunt chest trauma--a case report. AB - This report describes a case of traumatic incomplete rupture of the ventricular septum, a rare complication caused by blunt chest trauma. Although a serial ECG progressed its course similar to acute anteroseptal myocardial infarction in this case, there was little clinical clue of septal tear. The diagnosis was established by transthoracic echocardiography. The authors chose a conservative line of management rather than surgical repair for incomplete septal rupture because of the patent's stable clinical course and hemodynamic status. A sequence of echocardiography during a 32-day stay in the hospital showed no change in the extent of incomplete septal rupture, septal structure, systolic function, and shape of left ventricle and also obtained no evidence of shunting through the rupture. In conclusion, echocardiography is a useful investigation to make a diagnosis as well as for follow-up in case of incomplete ventricular septal rupture. A close follow-up of incomplete septal rupture with serial echocardiography should be performed, because several cases of delayed ventricular septal rupture following blunt chest trauma have been reported. PMID- 16703200 TI - LDL apheresis for cholesterol embolism following coronary artery bypass graft surgery--a case report. AB - A 76-year-old man without any prior history of abnormal urinalysis findings or renal insufficiency demonstrated mild renal dysfunction after coronary bypass graft surgery (CABG). Two months after CABG, pain and blueness in the toes (blue toe syndrome) appeared and, the serum creatinine level (S-Cr) increased from 1.2 to 2.0 mg/dL. On admission (3 months later), the urinary protein level was 0.5 g/day, white blood cell count 8,300/microL with eosinophils (Eo) 10.5%, S-Cr 2.1 mg/dL, and low-density lipoprotein (LDL) 106 mg/dL. Acute renal failure and blue toe syndrome due to a cholesterol embolism (CE) were diagnosed. Alprostadil 40 microg/day orally for 2 weeks and alprostadil 40 microg/day intravenously were used for 5 weeks, and Eo were 250/microL, S-Cr 2.5 mg/dL; however, blue toe syndrome gradually developed. At 8 weeks after admission, limaprost alfadex 30 microg/day orally was used for 3 weeks. However, the Eo gradually rose to 1,520/microL, S-Cr to 3.0 mg/dL, and LDL to 135 mg/dL, and LDL apheresis was therefore performed 20 times for CE. The data just after LDL apheresis was performed 10 times were as follows: Eo 1,120/microL, S-Cr 4.0 mg/dL, and LDL 89 mg/dL, and blue toe syndrome had disappeared. At 10 months after the first LDL apheresis, the Eo were 630/microL, S-Cr 2.9 mg/dL, and LDL 109 mg/dL. As a result, LDL apheresis was found to be beneficial for the treatment of CE with acute renal failure and blue toe syndrome after CABG. PMID- 16703201 TI - Acute myocardial infarction after a European hornet sting--a case report. AB - The authors briefly describe an 87-year-old man who experienced an acute ST elevation myocardial infarction in the context of an anaphylactic reaction caused by a European hornet (Vespa cabro linnaeus) envenomation. This unusual case highlights the potential cardiovascular complications associated with common insect stings such as those caused by hymenoptera. Thus, a thorough cardiovascular evaluation is essential in such cases since the vasoactive, inflammatory, and thrombogenic mediators of anaphylactic reactions may provoke myocardial ischemia. PMID- 16703202 TI - Postural compression of subclavian vein in droopy shoulder syndrome--a case report. AB - A patient with droopy shoulder syndrome (DSS) presented postural symptoms of venous compression of the left upper limb, in addition to those of traction at the brachial plexus. An antegrade phlebography clearly demonstrated impaired patency of the subclavian vein on neck extension. Recognition and management of the compromised venous drainage in patients with DSS is important in preventing vascular complications of the upper limbs. PMID- 16703203 TI - Rhythm disturbances in the form of atrial tachycardia in a female patient with polymyositis--a case report. AB - The presented case report describes a female patient suffering from polymyositis, in whom atrial tachycardia and depressed left ventricular function were diagnosed. Atrial tachycardia was confirmed by electrophysiological study, and the radiofrequency ablation failed to restore sinus rhythm. This case is an example of the tachy phase in brady-tachy syndrome. The patient is evaluated cardiologically on a regular basis with emphasis on bradycardia and asystole episodes because she was symptomatic for many years. PMID- 16703204 TI - Current management of extracranial carotid artery disease. AB - Stroke is the third most common cause of death in the United States. There are approximately 700,000 strokes/year; 80% are ischemic, and 20-30% of ischemic strokes are secondary to carotid disease. Carotid stenosis is traditionally treated by carotid endarterectomy (CEA). Multicenter, randomized, controlled trials have shown that surgery significantly reduces the risk of ipsilateral stroke in patients with severe symptomatic and asymptomatic carotid stenosis. Endovascular techniques for treating carotid stenosis have been developed over recent years. Carotid angioplasty and stenting (CAS) with cerebral protection has become an alternative to CEA for high-surgical-risk patients and the procedure of choice for stenoses inaccessible by surgery. In this review we summarize the existing data regarding the traditional state of management of extracranial carotid artery stenosis and compare these data to a critical analysis of the recent results of CAS. PMID- 16703205 TI - Rapid high-volume population screening for three major risk factors of future stroke: phase I results. AB - Three proximate risk factors for stroke are carotid stenosis, atrial fibrillation, and hypertension. Phase I of this prospective study was designed to establish the prevalence of these conditions among a population of health maintenance organization beneficiaries by using a rapid screening protocol in order to risk-stratify patients for appropriate management and subsequent cohort analysis. Patients at a tertiary care medical center were screened for stroke risk by using directed history, a 3-minute carotid "quick-scan'' protocol, an EKG lead II rhythm strip, and bilateral arm blood pressures. Patients with any abnormal result underwent specific diagnostic consultation with vascular surgery, cardiology, or primary care. These evaluations included formal carotid duplex ultrasound, 12-lead EKG +/- Holter monitor, and 5-day blood pressure check. Patients were then stratified into risk cohorts for appropriate management and future analysis of stroke incidence and outcomes. In 8 hours on a single day in October 2002, 294 patients (mean age 69) were screened. Combining history with results of screening and diagnostic tests, the overall prevalence of carotid stenosis was 6% (n = 17/294), atrial fibrillation 7% (n = 21/294), and severe hypertension 5% (n = 16/294). Fifty-nine patients (20%) screened positive for carotid stenosis by "quick-scan,'' and 29% (n = 17/59) of these had confirmed stenosis (>50%) in 1 or both arteries by formal duplex. The prevalence of confirmed carotid stenosis was 37% among those screening positive for 1 artery (odds ratio [OR] 14.6; p<0.001) and 75% among those screening positive for both (OR 74.7; p<0.001). Significant independent predictors of carotid stenosis by multivariate analysis included coronary artery disease or myocardial infarction, smoking, stroke or transient ischemic attack, male gender, and white race (all p<0.05). The prevalence of confirmed stenosis was 10% with any 3 predictors alone (OR 2.5; p<0.05), 31% with any 4 (OR 21.2; p<0.001), and 50% with all 5 (OR 46.5; p<0.001). Thirty-three patients (11%) were found to have a previously unidentified and untreated arrhythmia, and 12% (n = 4/33) of these had confirmed new atrial fibrillation; 158 patients (54%) had moderate hypertension and 16 (5%) had severe hypertension (>180/100). Overall, 82% (n = 242/294) of patients screened required additional diagnostic tests. Based on these results, 11% (n = 31/294) of patients were stratified as high risk, 64% (n = 188/294) as moderate risk, and 25% (n = 75/294) as low risk for future stroke. Rapid and efficient screening of a large population for stroke risk factors is feasible. The prevalence of undiagnosed, unsurveilled, and untreated carotid stenosis, atrial fibrillation, and severe hypertension is significant, as 75% of patients screened had 1 or more confirmed major risk factors for stroke. Phase II of this study will investigate the degree of stroke risk reduction possible with a multidisciplinary approach to early identification and aggressive treatment of these risks. PMID- 16703206 TI - Complications of diagnostic carotid/cerebral arteriography when performed by a vascular surgeon. AB - Carotid stenting has recently been considered as an alternative treatment to carotid endarterectomy for certain patients with carotid stenosis. Hence, performing carotid arteriography with minimal morbidity and mortality is essential. The purpose of this study was to audit complications of diagnostic carotid/cerebral arteriography performed by a vascular surgeon with experience in endovascular interventions. One hundred one consecutive patients underwent 4 vessel arch aortography with selective carotid, subclavian, and/or vertebral arteriography with use of the Seldinger technique. Demographic data, indications, procedure approach (transfemoral, brachial), number of arteries punctured, type of selective injection, contrast volume, and procedure time were analyzed. Minor complications were those that do not significantly alter the health or activity of the patient or require extra hospitalization or treatment. Other complications were defined as major complications. The technical success rate was 99% (100/101 patients). These included the following: 82 patients with right carotid artery, 82 with left carotid artery, 15 with right subclavian artery, 21 with left subclavian artery, 11 with right vertebral artery, and 17 with left vertebral artery (a total of 228 selective injections). Indications for procedures included the following: transient ischemic attack (TIA)/stroke symptoms in 66%, asymptomatic carotid stenosis in 22%, upper limb claudication in 4%, and vertebrobasilar insufficiency in 4%. Right femoral puncture was used in 79%, left femoral in 12%, and left brachial in 9%. The mean amount of contrast used was 101 cc (45-250 cc) and the mean procedure time was 46 minutes (22-132 minutes). There were 5 complications in the whole series: 3 major complications (3%), including 1 minor stroke (1%) with carotid injection, 1 TIA, and 1 major retroperitoneal bleeding; and 2 (2%) minor complications. The major complication rate in this series compares favorably to published rates of 5.7% to 9.1%. There was no association between complications and specific risk factors except for a longer catheterization time (66 minutes versus 45 minutes, p=0.011). Carotid/cerebral arteriography can be done safely by experienced vascular surgeons with minimal perioperative complications that compare favorably with what has been reported in the radiology literature. PMID- 16703207 TI - Barriers to endovascular aortic aneurysm repair: past experience and implications for future device development. AB - Despite improvements in endovascular aortic aneurysm repair (EVAR) devices and techniques, significant anatomic constraints still preclude successful EVAR in a large number of patients. The authors sought to identify the current barriers to EVAR and examine their evolution over time. Patients were evaluated for potential endovascular repair by computed tomography angiography (CTA) with or without supplemental conventional arteriograms. The patient population was separated into 2 groups (A and B) based on early and late time periods in the experience with EVAR, corresponding to the availability of various devices. Group A (early) consisted of the Guidant Ancure, Medtronic Talent, and AneuRx devices and comprised patients presenting between April 1997 through June 2000. Group B (late) consisted of the Medtronic AneuRx, Cook Zenith, Edwards Lifepath, Gore Excluder, and Endologix PowerLink devices and comprised patients presenting between July 2000 and December 2003. Patient demographics and anatomic reasons for rejection were recorded in a database for statistical analysis. In total, 547 patients were evaluated (463 men, 84 women). Of these, 346 patients (63%; 312 men, 34 women) were deemed suitable candidates for EVAR and 201 (37%; 151 men, 50 women) were rejected. There was no significant difference in the overall rate of rejection in the early vs the late time period (34% A, 41% B, p = 0.08), but the number of exclusion criteria per patient decreased over time; patients rejected for EVAR had an overall average of 1.6 exclusion criteria (Group A, 1.9; Group B, 1.2). The reasons for rejection did significantly change over time. Specifically, rejection on the basis of inadequate arterial access, presence of extensive iliac artery aneurysms, or an inadequate proximal neck decreased. A disproportionate number of women were excluded throughout the study: Group A, 56% of women compared to 30% of men (p = 0.0003); Group B, 63% of women compared to 36% of men (p = 0.0022). Women were more likely than men to have inadequate arterial access routes. In addition, patients with high operative risk were also more likely to be excluded from EVAR, a finding that persisted over time. Anatomic constraints continue to pose significant challenges to aortic endografting. Progress has been made in that technological advances have conquered some of the previous anatomic challenges, chiefly those of arterial access and treatment of concomitant iliac aneurysm disease. However, the overall rate of rejection for EVAR remains the same. The chief anatomic barriers continue to be the difficult aortic neck and management of branched vascular segments. PMID- 16703208 TI - Incidence and distribution of lower extremity deep vein thrombosis in rehabilitation patients: implications for screening. AB - Patients admitted to in-patient rehabilitation programs have an increased risk for developing deep venous thrombosis (DVT). However, the utility of screening for lower extremity DVT using duplex ultrasound in this high-risk population is not well characterized. The purpose of this study is to identify whether or not screening lower-extremity duplex exams are indicated in this high-risk population. Screening lower extremity duplex exams were performed on all patients admitted to the rehabilitation center at Mt. Sinai Hospital over a 3-year period. Charts were reviewed for patient age, gender, diagnosis, date of screening and follow-up duplex exams, presence and location of venous thrombosis at each duplex exam, history of anticoagulation, and medical DVT prophylaxis. The presence of DVT at screening, the location of DVT along the lower extremity, and the outcome of calf DVT were analyzed in terms of gender, underlying diagnosis, and history of DVT prophylaxis. Lower extremity DVT was detected in 34% of patients. Twenty three percent of patients had isolated calf vein thrombosis. Men were more likely than women to have DVT. Calf DVTs progressed in 3% of patients over an average follow-up of 2 weeks. The presence of DVT, its location along the lower extremity, and the outcome of calf vein DVT had no significant relationship to underlying diagnosis or history of prophylaxis. Screening duplex exams to detect lower extremity DVT in rehabilitation patients is useful. Screening altered management in 26% of patients, prompting either anticoagulation or repeat duplex exam. PMID- 16703209 TI - The application of split-thickness skin graft as an autogenous arterial conduit in a goat (Capra hircus) model. AB - Adequate autogenous vein is often the limiting factor in achieving a successful infrainguinal bypass. Attempts have been made to find alternative conduits; however, these alternatives have demonstrated inferior patency rates. We attempt to show that a split-thickness skin graft conduit provides a feasible autogenous arterial conduit. Neoconduits were prepared with an autogenous split-thickness skin graft (STSG) tubularized for a length of 5-6 centimeters with an appropriate caliber match to native artery. The deep dermal side of the graft was randomized to form either the external surface or the luminal surface. The neoconduit was placed as an interposition graft in the left common carotid artery. Grafts were studied in vivo with duplex ultrasonography and ex vivo by histopathology and immunohistochemistry. Feasibility study involved 4 animals with grafts harvested for study at 24 hours (n = 2) and 7 days (n = 2). Two subsequent groups were studied to evaluate 3-month (n = 8) and 6-month (n = 5) patency. All grafts (n = 4) in the feasibility phase of the study were patent at the time of harvest without evidence of aneurysmal degeneration. In the subsequent 8 goats, grafts with the deep dermal side forming the extraluminal surface (n = 4) had a propensity to ulcerate and rupture or to become aneurysmal (75%). The patency rate of these grafts at 6 weeks was 25%. In contrast, grafts with the deep dermal side forming the intraluminal surface (n = 4) demonstrated 75% patency at 6 weeks. Because of these results the remaining goats underwent placement of neoconduits with the deep dermal side forming the luminal surface. These grafts maintained a patency rate of 80% at 6 months. Neoconduits implanted with a diameter greater than 1.5 times the native arterial diameter became aneurysmal. Histopathology demonstrated neointimal formation in all grafts patent for longer than 7 days. Immunohistochemical staining for Factor VIII/von Willebrand's factor (vWF) was reactive in the endoluminal cells of these grafts. Immunohistochemical staining for a-smooth muscle actin demonstrated reactivity in conduits patent for greater than 1 month. Split-thickness skin may provide a feasible source for autogenous conduit in arterial reconstructions and warrants further study. Technical factors affecting patency include orientation of the deep dermal surface of the STSG and the diameter of the neoconduit at the time of implantation. PMID- 16703210 TI - Expert witness testimony: rules of engagement. AB - Overlooked in most legislative remedies to address the medical malpractice (MMP) crisis is stringent prohibition against the use of "junk science'' in the courtroom and defining the qualifications of an expert witness. Expert witnesses should be required to: (1) to disclose information materially related to the reliability of expert testimony. (2) Filing of a "Daubert brief'': a summary of the plaintiff's expert's opinion along with a resume outlining his/her knowledge, skill, experience, training and education, reputation in the field relevant to the litigation, and complete details of the methodology employed by the expert. (3) An oath or declaration that acknowledges the duty to disclose to the court all information known to the person to be material to the reliability of the expert witness. Rules of evidence: The "Frye test'' or the "general acceptance rule'' has been used by judges to exclude expert testimony unless it is "sufficiently established to have gained general acceptance in the particular field in which it belongs.'' Stricter standards need to be legislated at the state level. The mandatory use of scientific panels by judges in all MMP cases is needed. Legislative relief has been sought by the medical community to address the serious disconnect between negligence and MMP litigation. The use of junk science in the courtroom remains largely unaddressed in the judicial system. Medical societies and legislatures must act to define an expert witness and restrain plaintiff's attorneys from using junk science to influence juries. PMID- 16703211 TI - Entrapment of a cerebral embolic protection device--a case report. AB - Cerebral protection devices have significantly reduced the incidence of embolic events in patients undergoing carotid stenting. With the increasing availability of such devices an awareness of the potential complications associated with their deployment is essential. Here we report a patient with entrapment of a filter device within a carotid stent that required surgical removal. The mechanisms underlying this complication and measures to prevent its occurrence are discussed. PMID- 16703212 TI - Vertebral artery embolic protection via ipsilateral brachial approach during left subclavian artery angioplasty and stenting--a case report. AB - The purpose of this report is to explore angioplasty and stenting with cerebral embolic protection as a salvage procedure for a compromised carotid-subclavian bypass in the presence of antegrade vertebral artery flow. A 76-year-old woman with a carotid-subclavian bypass presented with graft infection. Failure of medical therapy to treat the infection prompted surgical removal of the graft. The native subclavian artery was still patent, but a severe complex proximal stenosis was present with antegrade flow into the left vertebral artery. Angioplasty and stenting of the subclavian artery was performed with cerebral protection achieved by positioning a FilterWire EX in the left vertebral artery via the left brachial artery approach. Deployment of a filter device in the vertebral artery via the brachial or radial approach can provide embolic protection without interfering with the subclavian artery stenting. The successful treatment of the subclavian artery enabled the complete removal of the infected graft without need for major vascular reconstruction. PMID- 16703214 TI - A true posterior tibial artery aneurysm--a case report. AB - Aneurysms of infrapopliteal arteries are rare. The etiology is usually traumatic, and most aneurysms are false. The English-language literature reports only 33 cases of infrapopliteal arterial true aneurysms, of which 8 involve the posterior tibial artery. The etiology of these lesions is unclear; a fibromuscular fibrodysplasia similar to ulnar aneurysm may be hypothesized, but traumatic, atherosclerotic, inflammatory, and other pathological processes are also probably involved. The natural history seems to be related to thrombosis and distal embolism more than to rupture. Surgical indications are debated. Aneurysm repair with a complete restoration of the blood flow through the affected artery is particularly challenging owing to the small size of the vessels, and ligation may be required. We report, to the best of our knowledge, the first case of an atherosclerotic posterior tibial artery true aneurysm successfully treated with aneurysmectomy and end-to-end direct reconstruction with a documented good long term patency. Clinical features, imaging findings, and surgical management are described; indications and treatments (open or endovascular) are discussed. PMID- 16703213 TI - Endograft repair of traumatic aortic transection in a 10-year-old--a case report. AB - Blunt abdominal aortic injury from trauma is extremely rare. It is generally from motor vehicular crashes resulting in dissection, intramural hematoma, or free rupture. Timely recognition and treatment of this injury are essential for a chance for survival. To the authors knowledge, this is the first reported case of successful management of this injury, with use of an intravascular stent-graft, in a child. Although open aortic repair has been the only modality used in children, intravascular stent-graft can be another available option in these injuries. PMID- 16703215 TI - Thoracic outlet syndrome as a disorder of neurogenic inflammation. PMID- 16703216 TI - Combined treatment with angiotensin-converting enzyme inhibitors and angiotensin II receptor blockers: a review of the current evidence. AB - Several studies have shown that angiotensin-converting enzyme inhibitors and angiotensin II receptor blockers are useful in the treatment of hypertension, cardiovascular disease, chronic heart failure, and some types of nephropathy. In this context, dual renin-angiotensin system blockade with both angiotensin converting enzyme inhibitors and angiotensin II receptor blockers may be more effective than treatment with each agent alone. Many clinical trials have demonstrated the beneficial effect of this combined treatment on proteinuria, hypertension, heart failure, and cardiovascular events. Moreover, these studies demonstrated that dual renin-angiotensin system blockade is generally safe and well tolerated. Long-term studies are under way to confirm these effects and also investigate the effectiveness of dual renin-angiotensin system blockade on cerebrovascular disease and prevention of type 2 diabetes mellitus. These studies are expected to define the optimal use of combination treatment in everyday clinical practice. This review considers the most important clinical trials that evaluated the effect of dual renin-angiotensin system blockade on blood pressure, heart failure, and renal function. PMID- 16703217 TI - Molecular aspects of ischemic heart disease: ischemia/reperfusion-induced genetic changes and potential applications of gene and RNA interference therapy. AB - Molecular biologic techniques have a variety of applications in the study of ischemic heart disease, including roles in elucidating cardiac genetic changes resulting from ischemia as well as in developing therapeutic interventions to treat ischemic heart disease. This review describes recent studies documenting genetic changes associated with myocardial ischemia and infarction as well as those investigating the safety and effectiveness of gene therapy for stimulating angiogenesis, protecting the heart against reperfusion injury, and treating heart failure. Also discussed are future research directions, including the potential use of RNA interference and combined stem cell therapy and gene therapy for the treatment of cardiovascular disease. PMID- 16703218 TI - Subcellular remodeling as a viable target for the treatment of congestive heart failure. AB - It is now well known that congestive heart failure (CHF) is invariably associated with cardiac hypertrophy, and changes in the shape and size of cardiomyocytes (cardiac remodeling) are considered to explain cardiac dysfunction in CHF. However, the mechanisms responsible for the transition of cardiac hypertrophy to heart failure are poorly understood. Several lines of evidence both from various experimental models of CHF and from patients with different types of CHF have indicated that the functions of different subcellular organelles such as extracellular matrix, sarcolemma, sarcoplasmic reticulum, myofibrils, mitochondria, and nucleus are defective. Subcellular abnormalities for protein contents, gene expression, and enzyme activities in the failing heart become evident as a consequence of prolonged hormonal imbalance, metabolic derangements, and cation maldistribution. In particular, the occurrence of oxidative stress, development of intracellular Ca2+ overload, activation of proteases and phospholipases, and alterations in cardiac gene expression result in changes in the biochemical composition, molecular structure, and function of different subcellular organelles (subcellular remodeling). Not only does subcellular remodeling appear to be intimately involved in the transition of cardiac hypertrophy to heart failure, the mismatching of the function of different subcellular organelles leads to the development of cardiac dysfunction. Although blockade of the renin-angiotensin system, sympathetic nervous system, and various other hormonal actions have been reported to produce beneficial effects on cardiac remodeling and heart dysfunction in CHF, the actions of various cardiac drugs on subcellular remodeling have not been examined extensively. Some recent studies have indicated that both the angiotensin-converting enzyme inhibitors and angiotensin receptor antagonists attenuate changes in sarcolemma, sarcoplasmic reticulum, and myofibril enzyme activities, protein contents, and gene expression, and partly improve cardiac function in the failing hearts. It is suggested that subcellular remodeling is an excellent target for the development of improved drug therapy for CHF. Furthermore, extensive studies should investigate the effects of different agents individually or in combination on reverse subcellular remodeling, cardiac remodeling, and cardiac dysfunction in various experimental models of CHF. PMID- 16703220 TI - Postconditioning markedly attenuates ventricular arrhythmias after ischemia reperfusion. AB - BACKGROUND: Brief periods of reocclusion (postconditioning) during early reperfusion reduce myocardial infarct size. Whether postconditioning has an effect on lethal ventricular arrhythmias independent of infarction in an in-vivo regional ischemia model is unknown. The purpose of this study was to determine if postconditioning limited reperfusion arrhythmias in a necrosis-free model. METHODS: Anesthetized rats were subjected to 5 minutes of proximal coronary artery occlusion; they were randomized to a control group (n = 15) that underwent reperfusion alone or a postconditioning group (n = 15) that received four cycles of 20 seconds reperfusion, 20 seconds reocclusion before final reperfusion. RESULTS: During the final reperfusion phase, ventricular arrhythmias occurred in 14 of 15 control rats and 8 of 15 postconditioning rats (P = .017). Ventricular tachycardia occurred in 10 of 15 control rats vs 4 of 15 postconditioning rats (P = .028). Control rats demonstrated 1.3 runs of ventricular tachycardia per minute vs 0.4 runs in postconditioning rats (P = .026). The average duration of ventricular tachycardia runs was 8.8 +/- 3.2 seconds in the control group vs 5.0 +/- 3.9 seconds in postconditioning rats (P = NS). CONCLUSION: This in-vivo study showed that postconditioning markedly attenuates ventricular arrhythmia after regional ischemia in a noninfarct model. PMID- 16703219 TI - Clopidogrel resistance: myth or reality? AB - Platelets have a central role in the development of arterial thrombosis and subsequent cardiovascular events. An appreciation of this complex process has made antiplatelet therapy the cornerstone of cardiovascular disease management. Dual antiplatelet therapy with aspirin and clopidogrel has been approved for the secondary prevention of cardiovascular events and is currently part of the postpercutaneous coronary intervention treatment regimen. However, subacute stent thrombosis continues to occur in 1% to 2% of patients despite dual antiplatelet therapy. Studies have shown interindividual variations in response to clopidogrel, where a cohort of patients seems to be resistant to the antithrombotic effects of clopidogrel. Furthermore, there is an apparent link between clopidogrel resistance and clinical outcomes. Currently, there is neither a universally accepted definition of clopidogrel resistance nor an agreement on the phenomenon's mechanism. This review highlights the origins of clopidogrel resistance, the current problems that exist with its definition, and discusses the future implications and relevant challenges it poses for the clinician. PMID- 16703222 TI - Post-transplantation lymphoproliferative disorder in heart and kidney transplant patients: a single-center experience. AB - BACKGROUND: Post-transplantation lymphoproliferative disorder (PTLD) after heart transplantation is a fatal complication, and standard treatment is either ineffective or too toxic. We have studied the incidence, clinical course, prognostic factors, and different treatment regimens pertaining to PTLD in 110 heart and 80 kidney transplant recipients. METHODS: Information was abstracted from chart review of 110 heart transplant recipients and 80 kidney transplant recipients between January 1989 and October 2002. We report 15 patients with PTLD, 6 patients received a heart transplant and 9 patients received a renal transplant. RESULTS: The overall incidence of PTLD was 8.9% (5.4% in heart and 13.7% in kidney transplant recipients). The average interval between transplantation and the diagnosis of PTLD in heart transplantation patients was 5.5 years, and their overall mean age was 44 years. The indications for transplantation were ischemic cardiomyopathy in 5 patients (1 patient received both heart and kidney transplants), glomerulonephritis in 6 patients, diabetes nephropathy in 2 patients, and polycystic disease in 2 patients. Six patients were diagnosed with early disease (<12 months), 7 with late onset (1 to 10 years), and 2 with very late onset (>10 years). Five patients had PTLD grade 2 (2 heart and 3 kidney transplants) and 10 patients had PTLD grade 3 (4 heart and 6 kidney transplants). Immunosuppressive treatment for PTLD patients consisted of cyclosporine, 73% (11/15); tacrolimus, 6.6% (1/15); prednisone, 100% (15/15); azathioprine, 80% (12/15); mycophenolate mofetil, 20% (3/15); murine monoclonal anti-human CD3 (OKT3), 7% (1/15); and anti-thymocyte globulin, 13% (2/15). PTLD developed in 11.5% of patients with primary Epstein-Barr virus infection and in 28.9% of patients with primary cytomegalovirus infection. Five patients received rituximab therapy, 5 had conventional chemotherapy, 3 had radiotherapy, 3 had reduction in immunosuppression, 2 had ganciclovir, 1 underwent surgery, and 1 patient died before receiving treatment. The mortality rate was 26.6%. The average interval between transplantation and the diagnosis of PTLD in heart transplant recipients was 5.5 years. The mortality rate was significantly higher in the control group than in the rituximab group. CONCLUSIONS: Caucasian race and male gender were independent risk factors for developing PTLD. Pretransplant cytomegalovirus seropositive status is a strong predictor of developing PTLD. Management of PTLD requires randomized controlled trials of various chemotherapeutic and antiviral drugs regimens. Treatment of PTLD with rituximab is a beneficial alternative with a favorable outcome. Patients in whom primary Epstein-Barr virus, cytomegalovirus, or hepatitis C infection develop after transplantation should be managed with heightened surveillance for the development of PTLD. Further randomized trials are needed to evaluate the efficacy of antiviral drugs, intravenous immunoglobulin, interferon, and prophylactic Epstein-Barr virus immunization strategies. PMID- 16703221 TI - Adaptive-outward and maladaptive-inward arterial remodeling measured by intravascular ultrasound in hyperhomocysteinemia and diabetes. AB - BACKGROUND: Coronary artery remodeling implies structural changes in the vessel wall in response to various pathophysiologic conditions. However, the classification of remodeling is unclear. We hypothesized that the adaptive, positive-outward remodeling is a reactive and compensatory response to the stress. The maladaptive negative-inward constrictive remodeling is a passive atherosclerotic condition in which the vessel becomes stiffer. METHODS: Patients with atherosclerotic lesions underwent intravascular ultrasound (IVUS) scans. The size of the vessels distal to and proximal to plaques were analyzed by IVUS. Diabetes was created in mice by an intraperitoneal injection of alloxan (65 mg/kg). To reduce remodeling, mice received ciglitazone, an agonist of peroxisome proliferators activated receptor-gamma (PPARgamma) in drinking water. After 8 weeks, atherosclerotic vessels were analyzed for collagen and elastin. RESULTS: IVUS data suggest an adaptive coronary arterial remodeling was a positive compensatory response to various pathologic stimuli; for example, with the deposition of atherosclerotic plaque, coronary arterial segments enlarged to maintain luminal area. This phenomenon was commonly observed during the initial phases of the development of atherosclerosis. However, negative coronary artery remodeling, or a decrease in vessel area with the formation of atherosclerotic plaque, was maladaptive and was associated with smoking, hypertension, hyperhomocysteinemia, diabetes mellitus, and also after percutaneous coronary interventions (restenosis). In diabetic mice, there was increased collagen and decreased elastin contents; however, treatment with ciglitazone ameliorated the decrease in elastin contents. CONCLUSION: Global enlargement of the coronary vascular tree occurs during pressure and volume overload associated with ventricular hypertrophic states such as athletic conditioning, hypertensive heart disease, and dilated cardiomyopathy. On the other hand, maladaptive coronary arterial remodeling occurs in patients with severe deconditioning, diabetes mellitus, after coronary artery bypass surgery, and in some instances, postintervention. PMID- 16703223 TI - Compliance by referring physicians with recommendations on heart failure therapy from a tertiary center. AB - BACKGROUND: We tested the assumption that general practitioners do not use medication as recommended and do not follow recommendations made by heart failure specialists. METHODS: The study included 193 stable patients referred to and subsequently seen at our heart failure clinic between 1995 and 2001 with an ejection fraction of 40% or less. They had 1906 visits (1661 with structured follow-up). Recommendations by heart failure specialists regarding medication influencing prognosis and adherence of referring physicians to them were recorded. RESULTS: Ninety-six percent of patients were taking angiotensin converting enzyme (ACE) inhibitor/angiotensin-II receptor blocker (ARB) when referred. The prescription rate of beta-blockers and spironolactone increased after publication of large trials (more than 80% in 2001). Doses were inappropriate for both ACE-inhibitors/ARBs (55% +/- 30% of target dose) and beta blockers (29% +/- 24%). Recommendations were followed closely for dose reduction/discontinuation of all drugs (82%) and start/uptitration of spironolactone (78%). Adherence was less (P < .01) for start/uptitration of ACE inhibitors/ARBs (62%) and beta-blockers (48%). Recommendations regarding single step start/uptitration were more successful (ACE-inhibitors/ARBs, 70%; beta blockers, 63%; single-step of both, 67%) than multiple-step uptitration (ACE inhibitors/ARBs, 48%; beta-blocker, 38%; P < .001). This was particularly true if recommendations concerned multiple-step uptitration of both drugs (25%, P < .001). There was no difference between general practitioners and cardiologists. CONCLUSIONS: Although penetration of drugs beneficial in heart failure was high upon referral, doses were lower than recommended, and recommendations regarding changes were insufficiently adhered to by the referring physicians. Simple, one step recommendations were better implemented and may be combined with direct prescriptions during control visits to guarantee adequate medical therapy. PMID- 16703224 TI - Circulating endothelial cells in venous blood as a marker of endothelial damage in chronic venous insufficiency: improvement with venoruton. AB - Damage to endothelial cells is common in vascular disorders and in reactions associated with transplantation. An elevated number of circulating endothelial cells indicates the extent of endothelial damage in a variety of disorders. In chronic venous insufficiency (CVI), the number of endothelial cells is abnormally increased, and this can be considered an important indication of endothelial damage. A group of 23 subjects with two levels of CVI (severe and very severe with previous ulcerations) with an increased endothelial cell count (seen by microscopy) was studied and treated for 4 weeks with oral Venoruton (0-[beta hydroxyethyl]-rutosides) (1 g/day) to evaluate the effects of treatment on the circulating endothelial cells in blood taken from a peripheral leg vein. The controls comprised two groups with comparable age and sex distribution, one of healthy individuals and one of CVI subjects. After 4 weeks, a significant decrease was noted in endothelial cells both in subjects with CVI and in those with very severe CVI with previous ulcerations. This study suggests that endothelial cells may play a significant role in venous disease, being both an indication of severe disease and a further problem in itself. The use of Venoruton appears to decrease the number of circulating endothelial cells. This suggests an important role of this compound in protecting the endothelium and offers new potentially important therapeutic options that are not limited only to venous disease. PMID- 16703225 TI - Comment: mortality benefit of no-load clopidogrel in COMMIT: not a surprise. PMID- 16703227 TI - Future directions in cardiac and vascular anesthesia: unanswered questions regarding variables controllable by anesthetic management. AB - The influence of anesthetic choices and techniques upon outcomes in cardiac surgery is difficult to ascertain because of the low incidence of morbid and fatal outcomes that are attributable to anesthetic management. This discussion summarizes several areas that have been addressed in clinical trials, including temperature management, intraoperative hemodynamics, and glucose control. PMID- 16703228 TI - Glucose tolerance testing in patients scheduled for cardiovascular surgery. AB - Diabetes and impaired glucose tolerance are largely underdiagnosed in patients with acute atherosclerotic events. This glucose-unawareness is an obstacle for aggressive treatment in these patients. It is suggested to check fasting glucose levels in patients scheduled for cardiovascular surgery and to give intensive insulin therapy perioperatively if fasting glucose levels are greater than 7.0 mmol/L (126 mg/dL). If fasting glucose levels are not elevated an oral glucose tolerance test should be considered so that unknown diabetes can be detected and treated. PMID- 16703229 TI - Repair of thoracic aneurysms, with special emphasis on the preoperative work-up. AB - The anesthetic approach to the patient with a thoracic aortic aneurysm depends on the urgency of repair. Symptomatic patients with leaking aneurysms require urgent intervention, and there is generally little time to perform more than the most basic preoperative assessment. For elective repair, however, one must consider nearly every organ system. Many of the specific issues are inherent to the underlying pathophysiology that has resulted in aneurysm formation, and some stem from the requirements of the surgical procedure itself. A thorough knowledge of the extent and location of the aneurysm, the functional status of the heart, and the coronary artery anatomy are critical. Most patients aged older than 40 years undergo coronary angiography preoperatively, as do younger patients with specific risk factors for myocardial ischemia. Respiratory failure is one of the most common sequelae of these procedures, and a thorough preoperative pulmonary work up is mandatory. Neurologic deficits are not uncommon postoperatively, and pre existing deficits in the central nervous system must be sought. Coagulopathy is common in the immediate postoperative period, and preoperative assurance of hemostatic competence is important. Computed tomography scans and magnetic resonance imaging are the mainstay of diagnosis, although the adjunctive use of echocardiography provides important information. Routine preoperative laboratory studies include complete blood count, chemistries, coagulation profile, and indices of renal function; an electrocardiogram, and chest radiograph. Close communication with the surgeon regarding the operative procedure, cannulation strategy (where applicable), and planned evoked potential monitoring is necessary to ensure appropriate perioperative management. Prophylactic antibiotics and antifibrinolytics are routine. PMID- 16703230 TI - Beta-blockade in the perioperative period: where do we stand after all the trials? AB - Perioperative myocardial infarction following noncardiac surgery is a complex process with a variety of proposed etiologic factors. Perioperative beta-blockade has been reported to reduce perioperative myocardial infarction and cardiac death, with possible direct effects on longer-term outcome, particularly after vascular surgery. Despite two high-profile studies that have pushed this topic into the mainstream of medicine, the number of patients studied and outcomes observed remains limited, especially for a therapy recommended for widespread adoption in millions of patients globally. Observational analyses, small meta analyses, and newer randomized trial data (primarily in abstract format) suggest the therapy is of benefit in high-risk patients, whereas in patients at intermediate or low risk, it is either mildly efficacious or neutral in effect. Adverse effects appear to be limited to the expected primary hemodynamic side effects of bradycardia and hypotension, although a suggestion of increased mortality has been reported in one observational analysis in the lowest-risk group. beta-Blockade may be associated with reduction in length of stay and reduced analgesic requirements, although these effects remain controversial. A single mega-trial being conducted outside of the United States with a target goal of 10,000 patients is ongoing and promises to provide definitive data within the next few years. Ongoing research into various etiologies of perioperative myocardial infarction and other medications with potential efficacy in this setting, including the important antiplatelet agents, must also be considered in developing recommendations for widespread adoption of perioperative beta blockade. PMID- 16703231 TI - Protecting the heart with cardiac medication in patients with left ventricular dysfunction undergoing major noncardiac vascular surgery. AB - Patients with left ventricular dysfunction who are undergoing major noncardiac vascular surgery are at increased risk of adverse postoperative events. We sought to evaluate whether perioperative medication use, including angiotensin converting enzyme (ACE) inhibitors, beta-blockers, statins, and aspirin, was associated with a reduced incidence of postoperative in-hospital mortality in these high-risk patients. The study enrolled 511 patients with left ventricular dysfunction (left ventricular ejection fraction <30%) who were undergoing major noncardiac vascular surgery. Cardiac risk factors and medication use were noted before surgery. Preoperative dobutamine stress echocardiography (DSE) was performed to identify patients with stress-induced myocardial ischemia. The end point was postoperative in-hospital mortality. Univariate and multivariate logistic regression analyses were performed to evaluate the relation between perioperative medication use and mortality. The mean age of the study population was 64 +/- 11 years, and 75% were men. Perioperative use of ACE inhibitors, beta blockers, statins, and aspirin was recorded in 215 (48%), 139 (27%), 107 (21%), and 125 patients (24%), respectively. Stress-induced myocardial ischemia occurred in 82 patients (16%). Sixty-four patients (13%) died. Perioperative use of ACE inhibitors (odds ratio [OR], 0.33; 95% confidence interval [CI], 0.12-0.91), beta blockers (OR, 0.03; 95% CI, 0.01-0.26), statins (OR, 0.06; 95% CI, 0.01-0.53), and aspirin (OR, 0.13; 95% CI, 0.03-0.55), was significantly associated with a reduced incidence of mortality, after adjusting for cardiac risk factors and DSE results. In conclusion, the present study showed that the perioperative use of ACE inhibitors, beta-blockers, statins, and aspirin is independently associated with a reduced incidence of in-hospital mortality in patients with left ventricular dysfunction who are undergoing major noncardiac vascular surgery. PMID- 16703232 TI - Volatile anesthetics and cardiac function. AB - All volatile anesthetics have been shown to induce a dose-dependent decrease in myocardial contractility and cardiac loading conditions. These depressant effects decrease myocardial oxygen demand and may, therefore, have a beneficial role on the myocardial oxygen balance during myocardial ischemia. Recently, experimental evidence has clearly demonstrated that in addition to these indirect protective effects, volatile anesthetic agents also have direct protective properties against reversible and irreversible ischemic myocardial damage. These properties have not only been related to a direct preconditioning effect but also to an effect on the extent of reperfusion injury. The implementation of these properties during clinical anesthesia can provide an additional tool in the treatment or prevention, or both, of ischemic cardiac dysfunction in the perioperative period. In the clinical practice, these effects should be associated with improved cardiac function, finally resulting in a better outcome in patients with coronary artery disease. The potential application of these protective properties of volatile anesthetic agents in clinical practice is the subject of ongoing research. This review summarizes the current knowledge on this subject. PMID- 16703233 TI - Intravenous anesthesia for the patient with left ventricular dysfunction. AB - Patients with heart failure have a diminished cardiac reserve capacity that may be further compromised by anesthesia. In addition to depression of sympathetic activity, most anaesthetics interfere with cardiovascular performance, either by a direct myocardial depression or by modifying cardiovascular control mechanisms. Etomidate causes the least cardiovascular depression. It is popular for induction of anesthesia in cardiac-compromised patients; however, it is not suitable for maintenance of anesthesia because it depresses adrenocortical function. Ketamine has a favorable cardiovascular profile related to central sympathetic stimulation and inhibition of neuronal catecholamine uptake. These counteract its direct negative inotropic effect. In patients with a failing myocardium, however, the negative inotropic effects may be unmasked, resulting in deterioration in cardiac performance and cardiovascular instability. Propofol is the most popular intravenous anesthetic for maintenance of anesthesia. It does have a negative inotropic effect, but the net effect on myocardial contractility is insignificant at clinical concentrations, probably because of a simultaneous increase in the sensitivity of the myofilaments to Ca2+. Propofol protects the myocardium against ischemia-reperfusion injury, an action derived from its antioxidant and free radical-scavenging properties as well as the related inhibition of the mitochondrial permeability transition pore. For intravenous anesthesia, propofol is always combined with an opioid. Opioids have relatively few cardiovascular side effects and, in particular, do not cause myocardial depression. Indeed, they are cardioprotective, with antiarrhythmic activity, and induce pharmacologic preconditioning of the myocardium by a mechanism similar to the inhalational anesthetics. PMID- 16703234 TI - The standard TEE examination: procedure, safety, typical cross-sections and anatomic correlations, and systematic analysis. AB - Multiplane transesophageal echocardiography (TEE) is a semi-invasive, very low risk imaging procedure utilizing the upper gastric and esophageal echo windows to image the heart with the use of high-frequency transducers. It allows for high spatial resolution and access to structures that are routinely not well seen by transthoracic echocardiography, such as the left atrial appendage, the thoracic aorta, and the pulmonary veins, or where transthoracic imaging may be insufficient to answer a specific question, such as the presence of small vegetations, endocarditic abscesses, abnormalities of the interatrial septum, and others. Another important indication is the ventilated patient or the patient in the operating room, where transthoracic echocardiography is unrewarding or impossible. A particular role has emerged for TEE for identifying candidates for mitral valve repair by providing detailed functional and morphologic information on the diseased valve and by checking intraoperatively the results of surgery. A set of cross-sections, defined by anatomic landmarks, has been compiled, which can be systematically sought, evaluated, and modified according to the clinical question in order to extract the maximal information from TEE. PMID- 16703235 TI - Quantification in echocardiography. AB - Until recently, more than 2200 Swan Ganz catheters were used annually in the operating rooms (OR) and intensive care unit (ICU) of the Catharina Hospital in Eindhoven, The Netherlands. After cardiologists who were specialists in echocardiography (ECHO) trained anesthesiologists in ECHO, the need for these catheters in cardiac and noncardiac surgery was reduced. Initially intended as a local teaching project, an ECHO teaching compact disk (CD) was produced during the training and distributed later worldwide, thanks to a positive review in a major anesthesiology publication. By reducing the number of Swan Ganz catheters, the hospital could finance and acquire two echocardiography machines for the OR and ICU. The availability of these machines resulted in a further reduction of the number of Swan Ganz catheters. However, the need for quantification (eg, measurements of cardiac output) remained. During the creation of the ECHO teaching CD, the idea was born to apply indicator-dilution principles on injected echo contrast. This study was performed in cooperation with the Signal Processing Department of the Eindhoven University of Technology. Advanced signal processing and modelling were used to develop algorithms to enable quantification of intrapulmonary blood volume, ejection-fraction, and flow from the transesophageal echocardiography approach. These quantitative measurements, which can be performed on an outpatient basis, may become a real asset in cardiology, anesthesiology, and intensive care. PMID- 16703236 TI - Advanced systolic and diastolic function: beyond the E- and A-wave. AB - The rationale and clinical use of Doppler echocardiography for the assessment of left ventricular diastolic dysfunction is described with focus on recent developments. Noninvasive measurement of elevated left atrial pressure can be performed with transthoracic and transesophageal echocardiography in daily practice and thus, can be used for clinical management in patients with heart failure. PMID- 16703237 TI - Evaluation of mitral regurgitation. AB - Transesophageal echocardiography (TEE) provides high-resolution images of the mitral valve apparatus, permitting detailed evaluation of its structure and function. Mitral valve evaluation is invaluable during mitral valve repair surgery, providing insight into the mechanism of valve dysfunction and formulation of a plan for repair. TEE provides immediate detection of inadequate repair after bypass and allows correction during the same operative setting. Intraoperative TEE is a valuable tool for improving patient outcome and offers the cardiovascular anesthesiologist a role in perioperative surgical decisions. PMID- 16703238 TI - Ischemic mitral regurgitation. AB - Ischemic mitral regurgitation is the regurgitation seen with structurally normal valve leaflets that occurs in approximately 20% of patients after myocardial infarction and 56% of patients with congestive heart failure caused by ischemic or nonischemic cardiomyopathy. The initiating event is an ischemic insult that results in remodeling of the left ventricle toward a more spherical shape and new wall motion abnormalities. These changes lead to annular dilation and subvalvular distortion that prevent the mitral leaflets from coapting and closing completely during the contraction phase. Treatment options include coronary revascularization, ring annuloplasty, valve repair and replacement, or left ventricle reconstruction by way of localized reshaping or resection. Pharmacotherapy, aimed at raising left ventricular pressure without increasing tethering or left ventricular volume, is included in the treatment options for improving ischemic mitral regurgitation. PMID- 16703239 TI - Echocardiography in the emergency department. AB - In the busy emergency department, time-effective and accurate diagnostic and prognostic evaluation is essential Although clinical examination remains the mainstay of appropriate evaluation, remaining diagnostic uncertainty may need to be resolved by use of additional techniques. Echocardiography appears very well suited to the unique environment in the emergency department. It is portable to the patient's bedside, can be performed in minimal time, and results are immediately available. This facilitates correct triage and disposition of complicated cases. Small, hand-carried echocardiography systems have been developed with full imaging and Doppler capabilities. Thus, availability of echocardiography is increasing. Because both image acquisition and interpretation are operator-dependent, realization of full diagnostic potential requires extensive training and post-training experience to maintain proficiency. PMID- 16703240 TI - Assessment of aortic dissection and hematoma. AB - Aortic dissection and aortic intramural hematoma are highly lethal diseases occurring mostly in patients with arterial hypertension or Marfan syndrome. Transesophageal echocardiography (TEE) is the diagnostic imaging procedure of choice in the unstable patient. Hallmarks of dissection are the presence of an intimal membrane, or flap, dividing a true and a false lumen, entry and reentry tears, periaortic hematoma, pericardial effusion or tamponade, and aortic regurgitation in dissection of the ascending aorta. TEE allows detection of involvement of the ascending aorta, which places the patient in an extremely high risk group and necessitates urgent operation. The accuracy of TEE is similar to computed tomography and magnetic resonance imaging, with the added benefit of being feasible at the bedside or in the operating room. Precautions have to be taken during TEE to avoid an abrupt rise in blood pressure. Aortic intramural hematoma is often a precursor of classic dissection or accompanies it and can also be diagnosed by TEE, although the diagnosis is frequently difficult. PMID- 16703241 TI - Evaluation of prosthetic heart valves by transesophageal echocardiography: problems, pitfalls, and timing of echocardiography. AB - Transesophageal echocardiography (TEE) is especially suitable for examination of prosthetic valves because of the proximity of the esophagus to the heart and absence of interference with lungs and ribs. This article reviews normal and abnormal morphologic characteristics of prosthetic valves such as spontaneous echocontrast, microbubbles, strands, sutures, vegetations or thrombus. Doppler echocardiographic characteristics of normal and pathologic prosthetic valve function and the management of prosthetic valve pathology is discussed. Physicians taking care of patients with prosthetic valves should be familiar with the characteristics of normal and abnormally functioning prosthetic valves. PMID- 16703242 TI - How to reliably detect ischemia in the intensive care unit and operating room. AB - Detection of myocardial ischemia in the perioperative period is important because it allows for intervention that may prevent progression of ischemia to myocardial infarction. Perioperative ischemia is also an important predictor of adverse cardiovascular outcomes. Patients should first be stratified according to their risk of having cardiovascular disease by identifying major, intermediate, and minor predictors of adverse cardiovascular outcome. Electrocardiographic (ECG) monitoring for ischemia is inexpensive and noninvasive, but may not be applicable to all patients and is not perfectly sensitive or specific. Modern operating room monitors can automate ST segment monitoring and be set to alarm if changes occur. Increases in central venous pressure and pulmonary artery pressure can be caused by myocardial ischemia, but have been shown to be very insensitive compared to ECG. Also, detection of these hemodynamic changes requires insertion of invasive monitoring devices. Transesophageal echocardiography can be used to detect myocardial ischemia by identifying changes in regional wall motion. These transesophageal echocardiography changes occur sooner and more frequently than ECG changes, but require greater knowledge and skill to properly interpret. PMID- 16703243 TI - The failing heart under stress: echocardiography is an essential monitoring tool in the intensive care unit. AB - Echocardiography has been evolving to play a pivotal role in hemodynamic management, both intraoperatively and at the bedside. A full assessment of hemodynamics necessitates the use of all of the options available on modern echocardiographs. This introductory review provides insight into three important issues of hemodynamic monitoring by echocardiography: evaluation of preloading conditions, assessment of systolic function, and contractility and estimation of afterload. Mastering these three features will help in a minimally invasive approach of hemodynamic instability. PMID- 16703247 TI - [Increase of musculoskeletal pain in Austria--what are we waiting for?]. PMID- 16703244 TI - Expression of ASK1-like genes in arrested stamens of female Silene latifolia plants. AB - Silene latifolia is a dioecious plant in which sex is determined by heteromorphic sex chromosomes. In female plants, stamen development is arrested before microspore mother cells are formed. In this study, we isolated four cDNAs (SlSKP1 1 to 4) encoding ASK1-like protein as expression markers to reveal when expression levels are reduced in arrested stamens of female flowers. Expression patterns of the SlSKP1 genes were analyzed by in-situ hybridization. We use the flower development classification of Grant et al. (in Plant J 6:471-480, 1994). SlSKP1 genes were highly expressed in primary parietal cells and primary sporogenous cells that develop into microspore mother cells in male flowers. Expression levels started to be reduced in the external stamens of the female flowers when stamen development was arrested at stage 7. Although microspore mother cells could not be developed in female flowers and SlSKP1 expression may be unnecessary in arrested stamens, SlSKP1 genes were still expressed in sporogenous cells of degenerated stamens at stage 8. Parietal cells stopped differentiating earlier than sporogenous cells in arrested stamens. These results suggest that not all types of cell are arrested simultaneously at a particular stage of stamen development during stamen suppression of S. latifolia. PMID- 16703248 TI - [Anal eczema -- an interdisciplinary diagnostic challenge]. PMID- 16703250 TI - Invasive carcinoma developing in condylomata gigantea Buschke-Lowenstein. PMID- 16703249 TI - Hereditary hemorrhagic telangiectasia: an update on clinical manifestations and diagnostic measures. AB - Hereditary hemorrhagic telangiectasia (HHT), also known as Rendu-Osler-Weber disease, is an autosomal dominant disorder of the fibrovascular tissue. It is characterized by the classic triad of (muco-)cutaneous telangiectases, arteriovenous malformations with recurrent epistaxis and hemorrhages, and inheritance. A wide variety of clinical manifestations in HHT have been described. In more than 90% of the patients, nosebleeds are the first predominant symptom, therefore ENT physicians often play a key role as far as diagnosis and management of the disease are concerned. In spite of recent diagnostic and therapeutic progress, a cure for this often burdening and handicapping disease is still not available. Apart from affecting the nose, arteriovenous malformations (AVMs) may also affect the skin, lungs, brain, liver and gastrointestinal tract. The two known genes that are implicated in HHT are endoglin (ENG) located on chromosome 9q33-q34 and activin-receptor-like kinase (ALK1) located on chromosome 12q13. Mutations of ENG are observed in HHT type 1 with an incidence up to 40% for pulmonary AVMs, whereas mutations of ALK1 are observed in HHT type 2 with an incidence of only 14% for pulmonary AVMs, which clinically distinguishes these two types of mutation. The emphasis of this paper is mainly on the clinical manifestation, molecular genetics and diagnosis of HHT, taking account of current literature on HHT in order to better understand the complexity of the disease. Recent therapeutic options in the treatment of HHT have been omitted from this paper as they are subject of a following paper. HHT is more common than previously thought and shows a broad range of different clinical organ manifestations that can be sources of substantial morbidity and mortality, making HHT a continuing challenge for many sub-specialties where interdisciplinary diagnostic screening is mandatory in the management of the disease. PMID- 16703251 TI - [Prevalence of self-reported musculoskeletal pain in the Austrian population]. AB - INTRODUCTION: The objective of this study was to document the prevalence of musculoskeletal pain in the Austrian population, to analyze the specific pain of affected individuals and to compare the characteristics of the group of persons with pain to those without pain. METHODOLOGY: Representative samples of the Austrian general population were selected by the "quota procedure". 500 persons, aged 15 years and older were questioned in face-to-face interviews. RESULTS: 36.4% of all questioned individuals had experienced pain in the musculoskeletal system within the last three weeks, primarily in the region of the spine. Most of the very severe pain was located in the neck area (22%). The groups of persons with and without pain differed significantly in age, occupation, occupational group, region of residence within Austria, and family status. In a regressive analysis higher age remained the only significant variable. In seeking treatment, of significant importance were higher pain intensity and higher education level, and longer time since pain onset. CONCLUSION: The high prevalence of musculoskeletal pain in the Austrian population, particularly in the advanced age group not only requires targeted and high quality treatment of the affected persons, but also attention to appropriate preventive measures. PMID- 16703252 TI - Etiologic and causative factors in perianal dermatitis: results of a prospective study in 126 patients. AB - BACKGROUND: Perianal dermatitis is probably the most common cutaneous disorder of the genitoanal area. Studies on the epidemiology of causative factors are rare. METHODS: Over a 4-year period we prospectively studied 126 patients with a presumptive diagnosis of anal eczema. The diagnostic algorithm comprised medical history, inspection, microbiology, laboratory chemistry, patch tests, proctoscopy, and biopsy if appropriate. RESULTS: The age range was 7-82 years and the majority of patients were male (57.1%). Periods of anal symptomatology ranged from 6 days to 120 months and most of the patients (51.6%) had complaints for more than 12 months. The clinical diagnosis in 68 patients (54%) was: intertrigo/candidiasis (42.9%), atopic dermatitis (6.3%), pruritus ani (5.6%), psoriasis (3.2%), skin atrophy from steroid use (2.4%), lichen sclerosus et atrophicus (n = 2), herpes simplex (n = 1), and condylomata acuminata (n = 1). Contact eczema was suspected in 58 patients (46%), but 25 of these (43.1%) showed no contact sensitization. CONCLUSION: The majority of patients with symptoms of anal eczema suffer from intertrigo/candidiasis, and relevant, causative contact sensitization may be found in only some of them. Patch-testing is a valuable investigative tool only when the patients' own products are included in the test series. Most patients suffer from their perianal complaints for more than 12 months, therefore diligent evaluation is warranted. PMID- 16703253 TI - Adjuvant laser acupuncture in the treatment of whiplash injuries: a prospective, randomized placebo-controlled trial. AB - Following introduction of the compulsory use of seat belts in cars, whiplash injuries of the cervical spine have become common in everyday practice. Current treatment approaches lead to resolution of the symptoms within a short time in most cases but cannot prevent a small proportion of patients developing persistent health problems. The effects of adjuvant treatment with laser acupuncture on the acute symptoms and the results one year after the injury were studied in this prospective, randomized, placebo-controlled single-blind study. One group of patients (n = 23) were treated with laser acupuncture (5 mW HeNe laser on 22 acupuncture points for 15 s each) plus cervical collar and a combination of paracetamol and chlormezanone; a second group (n = 22) received the same treatments but with the use of a placebo laser. The treatment was given three times per week until the patient was asymptomatic. No statistically significant advantage of the laser acupuncture treatment was found in the acute phase (mobility in all three planes, duration of pain and duration of use of a cervical collar) or the chronic phase (drug use and the incidences of chronic recurrent problems such as myofascial pain, headaches, vertigo and tinnitus). CONCLUSION: Adjuvant laser acupuncture with a 5 mW HeNe laser and an irradiation time of 15 s appears to be ineffective in the management of whiplash injuries. PMID- 16703254 TI - Supply of R-alpha-lipoic acid and glutamine to casein-fed mice influences the number of B lymphocytes and tissue glutathione levels during endotoxemia. AB - BACKGROUND: An overwhelming production of reactive oxygen species concomitant with a decrease in antioxidative capacity plays an important role in modulation of the immune system in critically ill patients. The purpose of this study was to assess the influence of a combined oral supply of the antioxidants R-alpha-lipoic acid (LA) and glutamine (GLN) on the immunity of endotoxemic mice, with a special focus on tissue glutathione levels. METHODS: Female Balb/c mice were fed diets enriched with GLN (3 g/100 kcal), LA (0.74 mg/100 kcal), a combination of GLN and LA, or an isocaloric and isonitrogenous control diet for 10 days. On day 7, the mice were challenged intraperitoneally with 25 microg lipopolysaccharide. Seventy two hours later, the number and phenotype of lymphocytes in Peyer's patches (PP) and spleen of the endotoxemic mice were measured. In addition, glutathione levels were determined in the small intestine, spleen and liver. RESULTS: In PP only the combined supply of GLN and LA significantly increased the total cell yield (+19%), which was predominantly due to an increased number of B cells. In the spleen, both LA (+17%) and the combination of GLN and LA (+22%) were able to enhance total cell yield. The glutathione content of the small intestine was increased by feeding LA alone, whereas in the spleen GLN plus LA was most effective. CONCLUSION: Supplying combined GLN and LA to endotoxemic mice is effective in selectively increasing the number of systemic and intestinal B lymphocytes. Furthermore, LA augmented the level of the main intracellular antioxidant glutathione in the small intestine. On the basis of these data we recommend investigation of the effects of LA and GLN supplementation in patients with sepsis. PMID- 16703255 TI - Acute effects of hemodialysis on lung function in patients with end-stage renal disease. AB - Impaired lung function in hemodialysis patients may be caused by an underlying pulmonary disease; however, the impact of uremia and the effects of dialysis treatment are not well understood. Our investigation aimed to characterize the acute effects of bicarbonate hemodialysis using membranes differing in biocompatibility on various parameters of lung function in unselected uremic patients maintained on regular hemodialysis. Fourteen clinically stable hemodialysis patients without acute lung disease were included in the study. Restrictive lung disease was present in eight of 14 cases and obstructive lung disease in one patient. A cellulose dialyzer membrane and a synthetic high-flux dialyzer membrane were each tested twice (two sessions one week apart). Spirometry (VCmax, FEV1, FEF(25-75%), PEF) was carried out before and after hemodialysis. Resistance was determined with the interrupter technique and with the impulse oscillation system (R5Hz, R20Hz) before, during and after hemodialysis. Our comparative investigation of two dialyzer membranes found that bioincompatibility of dialysis had no acute adverse effects on lung function in our heterogeneous population of dialysis patients. None of our patients experienced bronchoconstriction or aggravation of obstructive lung disease as a result of poor biocompatibility of the dialyzer membrane. Spirometric data and resistance measurements by two different methods showed no relevant changes during the dialysis procedure. There was no correlation between lung function parameters and interdialytic changes in body weight or duration on hemodialysis. Regardless of the membrane used, the hemodialysis procedure does not acutely affect lung function in uremic patients on maintenance hemodialysis. Hemodialysis is a safe procedure even in uremic patients with pre-existing lung disease. PMID- 16703256 TI - [Sentinel lymph node in oral and oropharyngeal epithelial tumors]. AB - Carcinomatous metastases in regional lymph nodes worsen substantially the prognosis of patients with oral cavity and oropharyngeal cancer. Due to the high probability of occult metastasis (about 30%), during surgical resection of the primary tumor usually also elective dissection of lymph nodes is performed. Opinions on the extent of the elective neck dissection still differ, whereas selective dissection increasingly gains in importance. The aim of selective dissections, based on the predictability of formation of metastases, is the identification and exstirpation of the sentinel lymph node. In this prospective study the applicability of the concept of the sentinel lymph node in patients with oral cavity and oropharyngeal cancer was analysed. 12 patients with oral cavity and orophangeal cancer, staging T1-T3, all N0 (examined by palpation and sonography) were included. The localization of the sentinel(s) was determined preoperatively by radioisotope (Tc Nanocolloid). Sentinel(s) were identified first with a gamma probe (Neoprobe 2000); we then injected methylene blue into the peritumoral area for easier detection of the sentinel(s). The sentinels were removed and sent for frozen section examination. Regardless of the findings of the frozen section examination modified dissection was carried out. Later we compared frozen sections with paraffin microtome sections of sentinel(s) and of other exstirpated neck lymph nodes. We could identify the sentinel lymph node in all patients, in 6/12 patients we found several sentinels. If sentinels were not affected by tumor cells, there were no metastases in the downstream neck lymph nodes either. If in the sentinel lymph nodes no metastases can be determined, eliminating the environment alone could be sufficient. However, this assumption requires verification in a larger patient group. PMID- 16703257 TI - Wegener's granulomatosis of the breast. AB - Wegener's granulomatosis is a multisystem disorder characterized by necrotizing granulomatous inflammation and vasculitis of small vessels and can affect any organ system. The most common sites of involvement are upper and lower respiratory tracts, and kidneys. Breast involvement is unusual and very rare. We report a case of breast Wegener's granulomatosis in a 32-year-old woman who presented with pulmonary lesions and palpable masses in the left breast. Mammography showed multiple, sharply delineated nodules without microcalcifications. Ultrasonography revealed multiple hypoechoic solid lesions, some of them with anechoic areas of necrosis. Computed tomography showed multiple nodules. Histopathology of excision biopsy specimens of breast lesions revealed necrotizing granulomatous material consistent with Wegener's granulomatosis. Twenty reports of breast involvement in this rare disease were found in the literature; however, the respective ultrasonographic and computed tomography findings have not hitherto been described. PMID- 16703258 TI - Axillary lymph node metastasis in recurrence of papillary thyroid carcinoma: a case report. AB - Papillary thyroid cancer usually metastasizes to regional lymph nodes and to distant sites such as lungs and bones. We report a case of axillary lymph node metastasis as a result of recurrence of papillary carcinoma in a 62-year-old woman with papillary thyroid cancer extending locally beyond the thyroid capsule. Six years after initial surgical treatment, a lymph node metastasis in the left axillary region was diagnosed with positron tomography. To our knowledge, only one previous case of confirmed axillary metastasis of thyroid cancer has ever been reported. These two cases provide some evidence that thyroid carcinoma may exceptionally spread to axillary lymph nodes. Hypotheses that may account for such unusual localization include hematogenous dissemination or retrograde dissemination to regional lymphatic channels. Thus, when recurrence of thyroid carcinoma is considered, careful clinical examination of the axilla is recommended. Furthermore, thyroid carcinoma must be considered in the differential diagnosis of an axillary mass, especially when breast cancer is ruled out. PMID- 16703260 TI - Amino acid starvation induced autophagic cell death in PC-12 cells: evidence for activation of caspase-3 but not calpain-1. AB - While the apoptotic and necrotic cell death pathways have been well studied, there lacks a comprehensive understanding of the molecular events involving autophagic cell death. We examined the potential roles of the apoptosis-linked caspase-3 and the necrosis/apoptosis-linked calpain-1 after autophagy induction under prolonged amino acid (AA) starvation conditions in PC-12 cells. Autophagy induction was observed as early as three hours following amino acid withdrawal. Cell death, measured by lactate dehydrogenase (LDH) and 3-(4,5-dimethylthiazol-2 yl)-2,5-diphenyltetrazolium bromide (MTT) assays occurred within 24 h following starvation and was accompanied by an upregulation in caspase-3 activity but not calpain-1. The cell death that occurred following AA starvation was significantly alleviated by treatment with the autophagy inhibitor 3-methyl adenine but not with the broad spectrum caspase inhibitors. Thus, this study demonstrates that 3 methyladenine-sensitive autophagic cell death due to AA starvation in PC-12 cells is mechanistically and biochemically similar to, yet distinct from, classic caspase dependent apoptosis. PMID- 16703261 TI - The extrinsic and intrinsic apoptotic pathways are differentially affected by temperature upstream of mitochondrial damage. AB - It is well known that mild hypothermia prevents neuronal cell death following cerebral ischemia, although it can also cause apoptosis in other cell types. Thus, incubation at room temperature (RT) has been shown to induce apoptosis in hematopoietic cells, including Jurkat T leukemia cells. To further understand the apoptotic events that can be activated at RT, we compared the induction of apoptosis by several apoptotic insults in Jurkat cells stimulated at 37 degrees C or RT. Retinoid-related molecules, which induce apoptosis via the intrinsic pathway, failed to induce apoptosis when cells were treated at RT, as determined by various apoptotic parameters including cytochrome c release and activation of caspase 3. In contrast, most apoptotic events were enhanced by lower temperatures when cells were stimulated with anti-Fas antibody via the extrinsic pathway. Ultraviolet radiation produced partial effects at RT, correlating with its capacity to activate both pathways. Our results indicate that the core caspase machinery is operational under mild hypothermia conditions. Experiments using purified recombinant caspases and cell-free assays confirmed that caspases are fully functional at RT. Other hallmark events of apoptosis, such as phosphatidylserine externalization and formation of apoptotic bodies were variably affected by RT in a stimulus-dependent manner, suggesting the existence of critical steps that are sensitive to temperature. Thus, analysis of apoptosis at RT might be useful to (i) discriminate between the extrinsic and intrinsic pathways in Jurkat cells treated with prospective stimuli, and (ii) to unravel temperature-sensitive steps of apoptotic signaling cascades. PMID- 16703262 TI - Essential role of c-Jun-NH2-terminal kinase on synergy induction of apoptosis by TRAIL plus ADM in ADM resistant MCF-7/ADM cells. AB - Combined treatment modalities using tumor necrosis factor related apoptosis inducing ligand L (TRAIL) and cytotoxic drugs revealed highly additive effects in some tumor cell lines. Little is known about the efficacy and underlying mechanistic effects of the modalities in chemoresistant tumor cells. The purpose of this study is to investigate the possible role of JNK in the synergistic effect in Doxorubicin (Adriamycin, ADM) resistant MCF-7/ADM cells. Here we showed that the JNK pathway was activated slightly by TRAIL in MCF-7/ADM cell lines and was enhanced by the combination of the two treatments. Inhibition of JNK activity by transfection with dominant-negative JNK blocks TRAIL plus ADM induced apoptosis significantly, and selective stimulation of the JNK pathway sensitizes ADM resistant breast cancer cells to ADM and TRAIL co-treatment through activation of mitochondria-regulated apoptotic pathway. We conclude that the JNK pathway plays an important role in mediating TRAIL plus ADM induced-apoptosis in breast cancer cells. PMID- 16703263 TI - Differential protective effects of palmitoleic acid and cAMP on caspase activation and cell viability in pancreatic beta-cells exposed to palmitate. AB - Saturated and mono-unsaturated fatty acids exert differential effects on pancreatic beta-cell viability during chronic exposure. Long chain saturated molecules (e.g. palmitate) are cytotoxic to beta-cells and this is associated with caspase activation and induction of apoptosis. By contrast, mono-unsaturated fatty acids (e.g. palmitoleate) are not toxic and can protect against the detrimental effects of palmitate. In the present study, we show that the protective actions of palmitoleate in BRIN-BD11 beta-cells result in attenuated caspase activation following exposure to palmitate and that a similar response occurs in cells having elevated levels of cAMP. However, unlike palmitoleate, elevation of cAMP was unable to prevent the cytotoxic actions of palmitate since it caused a diversion of the pathway of cell death from apoptosis to necrosis. Palmitoleate did not alter cAMP levels in BRIN-BD11 cells and the results suggest that a change in cAMP is not involved in mediating the protective effects of this fatty acid. Moreover, they reveal that attenuated caspase activation does not always correlate with altered cell viability in cultured beta-cells and suggest that mono-unsaturated fatty acids control cell viability by regulating a different step in the apoptotic pathway from that influenced by cAMP. PMID- 16703264 TI - Higenamine reduces apoptotic cell death by induction of heme oxygenase-1 in rat myocardial ischemia-reperfusion injury. AB - Pharmacological modulation of heme oxygenase (HO) gene expression may have significant therapeutic potential in oxidant-induced disorders, such as ischemia reperfusion (I/R) injury. Higenamine is known to reduce ischemic damages by unknown mechanism(s). The protective effect of higenamine on myocardial I/R induced injury was investigated. Ligation of rat left anterior descending coronary artery for 30 min under anesthesia was done and followed by 24 h reperfusion before sacrifice. I/R-induced myocardial damages were associated with mitochondria-dependent apoptosis as evidenced by the increase of cytochrome c release and caspase-3 activity. Administration of higenamine (bolus, i.p) 1 h prior to I/R-injury significantly decreased the release of cytochrome c, caspase 3 activity, and Bax expression but up-regulated the expression of Bcl-2, HO-1, and HO enzyme activity in the left ventricles, which were inhibited by ZnPP IX, an enzyme inhibitor of HO-1. In addition, DNA-strand break-, immunohistochemical analysis, and TUNEL staining also supported the anti-apoptotic effect of higenamine in I/R-injury. Most importantly, administration of ZnPP IX inhibited the beneficial effect of higenamine. Taken together, it is concluded that HO-1 plays a core role for the protective action of higenamine in I/R-induced myocardial injury. PMID- 16703265 TI - Involvement of DNA replication in ultraviolet-induced apoptosis of mammalian cells. AB - Exposure of cells to ultraviolet (UV) light damages the genome and the persistence of DNA lesions triggers apoptosis in mammalian cells. RNA transcription blockage by DNA damage is believed to be implicated in signaling for UV-induced apoptosis, but the role played by DNA replication in this process is still unclear. To address this point, we have employed the DNA polymerase inhibitor aphidicolin in UV-irradiated wild-type and XPB-mutated Chinese hamster ovary cells. The data obtained with synchronized cells indicate that induction of apoptosis by UV light is independent of the cell cycle phase. Nevertheless, cells treated with aphidicolin after UV exposure showed a significant prevention of apoptosis induction when compared to proliferating cells. These results were observed in both DNA-repair proficient and deficient cells, indicating that the prevention of apoptosis by aphidicolin is independent of the cells' ability to repair the photolesions caused by UV. Taken together, these data suggest that replication of damaged DNA also leads to critical events signaling for UV-induced cell death. PMID- 16703267 TI - Identification of potential drug-related problems in the elderly: the role of the community pharmacist. AB - OBJECTIVE: The high prevalence of multiple drug use combined with age-related changes in pharmacokinetics and pharmacodynamics makes older adults more vulnerable to drug-related problems (DRPs). This pharmacy-based study was performed to identify potential DRPs from prescription records of the elderly and the role of the pharmacist in this process. METHOD: The study was performed from June 2002 to February 2003 in 16 community pharmacies in the Netherlands. Medication assessment of elderly patients aged 65 and over using six or more drugs concomitantly took place on the date of inclusion. Ten types of potential DRPs, grouped into three categories, were determined. The three groups were patient-related, prescriber-related or drug-related potential DRPs. We looked at the occurrence, nature and determinants of differential potential DRPs. RESULTS: The mean number of prescriptions per patient was 8.7. In total 3.9 potential DRPs per elderly person were identified. The distribution of the potential DRPs over the three categories was: patient related 4.7%, prescriber related 55.7% and drug related 39.6%. Use of NSAIDs (OR 29.9; 95% CI 4.1-219) and digoxin (OR 15.7; 95% CI 4.9-50.5) were associated with the highest risk for potential DRPs. CONCLUSION: In this vulnerable group of elderly patients potential DRPs frequently occur. Community pharmacists can play an important role in the identification, assessment and prevention of potential DRPs in the elderly. It is useful to investigate which part of potential DRPs can be avoided by the intervention of the community pharmacist in collaboration with the prescriber and the patient. PMID- 16703268 TI - Regional influences on the dispensing of glucose test strips in Dutch community pharmacies. AB - BACKGROUND: Pharmacy practice guidelines promote the role of community pharmacies in self-monitoring of blood glucose. However, variations between Dutch pharmacies exist in the proportion of patients to whom test strips are dispensed. OBJECTIVE: To assess whether variations between community pharmacies in dispensing of blood glucose test strips can be explained by differences in patient characteristics and the region in which the pharmacy is located. SETTING: PHARMO-Record Linkage System containing drug dispensing histories from 40 community pharmacies of about 450,000 patients in the Netherlands. METHOD: We performed a retrospective cohort study. Data on prescription of all drugs and medical aids between 1991 and 2001 were extracted for all new users of antidiabetic drugs. Patients were excluded if the dispensing history did not allow classification of the type of diabetes or if the dispensing pharmacy could not be determined. The data were analysed using a Cox proportional hazard model. MAIN OUTCOME MEASURE: Time to first test strips dispensed. RESULTS: We identified 8,233 starters of antidiabetic drugs. During a median follow-up of 2.1 years, 20% of the patients were dispensed test strips at least once. Community pharmacy was significantly associated with the dispensing of test strips after adjustment for patient characteristics. This association was less apparent when stratified for the geographical location of the pharmacy. CONCLUSION: Community pharmacy is an independent determinant of the start of use of test strips. Differences in dispensing of test strips between pharmacies are dependent on geographical region. This suggests that implementing practice guidelines for diabetes care in community pharmacy requires different approaches in different regions. PMID- 16703269 TI - Prevalence of self-medication in rural areas of Portugal. AB - OBJECTIVES: To study the prevalence of self-medication among pharmacy customers in rural areas of Portugal, to assess possible predictors of self-medication and to find out whether there was a seasonal dependence in the purchase of drugs for self-medication. METHOD: A cross-sectional study during four different periods of a year was conducted. Community pharmacies of rural areas of Portugal were invited to participate and pharmacists were asked to recruit one person every hour during the opening hours and administer a questionnaire. Drugs dispensed were classified according to the Anatomical Therapeutic Chemical Classification system up to the second level. MAIN OUTCOME MEASURE: Prevalence of self medication is defined as the percentage of patients acquiring a medicine that was not prescribed (written) or recommended (orally) by a physician. RESULTS: The prevalence of self-medication was 21.5%. Main therapeutic groups acquired for self-medication were "other alimentary tract and metabolism products" (A16; proportion acquired for self-medication= 75.0%), "throat preparations" (R02; 74.7%), "antiemetics and antinauseants" (A04; 70.0%), "cough and cold preparations" (R05; 56.5%), and "nasal preparations" (R01; 50.0%). Variables found to be predictors of self-medication were age, type of health professional or person consulted when a mild health problem occurred, time elapsed since last visit to the physician and time waited between setting an appointment and the actual visit. Seasonality seemed to occur for only "cough and cold preparations", for "dermatologicals" and for "anti-inflammatory and anti-rheumatic products". CONCLUSION: In rural Portugal about one fifth of the pharmacy customers engaged in self-medication. However, further research should be made to address appropriateness of self-medication. PMID- 16703270 TI - Comparison of the effects of melatonin and pentoxifylline on carbon tetrachloride induced liver toxicity in mice. AB - The purpose of the study was to determine whether along and in combination melatonin (MLT) and pentoxifylline (PTX) exerted beneficial effects on histopathological changes and changes in oxidant and antioxidant systems in liver caused by CCl4-induced liver toxicity in mice. Mice were randomly divided into six groups: control, olive oil, toxicity, MLT, PTX, PTX+MLT. MLT 10 mg/kg/day, PTX 50 mg/kg/day, and the same individual doses in MLT+PTX combination were given intraperitoneally to mice for 7 day. CCl4 0.8 mg/kg/day was administered on the 4th, 5th, and 6th days of therapy in all groups except the control and olive oil groups. In the toxicity group, increased concentrations of malondialdehyde (MDA) and lipid hydroperoxides (LOOH) and decreased glutathione peroxidase (GSH-Px) and catalase (CAT) activities were found compared to the control and olive oil groups (p < 0.05). Compared to the toxicity group, both the PTX group and the PTX+MLT group had decreased MDA and LOOH levels, whereas MLT reduced only LOOH levels (p < 0.01). MLT, PTX and MLT+PTX increased the GSH-Px and CAT activities compared to the toxicity group (p < 0.05). MLT increased CAT activity compared to PTX and MLT+PTX (p < 0.05). Superoxide dismutase enzyme activity did not change in any group (p < 0.05). Histopathologically, ballooning, degeneration, apoptosis, and bridging necrosis were seen in the toxicity group. MLT, PTX and MLT+PTX decreased the apoptosis and bridging necrosis (p < 0.01), and PTX and MLT+PTX decreased balloon degeneration compared to the toxicity group (p < 0.01). These results indicate that administration of PTX and MLT alone and in combination before onset of liver toxicity might prevent the oxidative damage by reducing oxidative stress and increasing antioxidant enzyme levels. PMID- 16703271 TI - Improvement of human dendritic cell culture for immunotoxicological investigations. AB - A toxic injury such as a decrease in the number of immature dendritic cells caused by a cytotoxic effect or a disturbance in their maturation process can be responsible for immunodepression. There is a need to improve in vitro assays on human dendritic cells used to detect and evaluate adverse effects of xenobiotics. Two aspects were explored in this work: cytotoxic effects of xenobiotics on immature dendritic cells, and the interference of xenobiotics with dendritic cell maturation. Dendritic cells of two different origins were tested. Dendritic cells obtained either from umbilical cord blood CD34(+) cells or, for the first time, from umbilical cord blood monocytes. The cytotoxicity assay on immature dendritic cells has been improved. For the study of the potential adverse effects of xenobiotics on the maturation process of dendritic cells, several parameters were selected such as expression of markers (CD86, CD83, HLA-DR), secretion of interleukins 10 and 12, and proliferation of autologous lymphocytes. The relevance and the efficiency of the protocol applied were tested using two mycotoxins, T-2 toxin and deoxynivalence, DON, which are known to be immunosuppressive, and one phycotoxin, domoic acid, which is known not to have any immunotoxic effect. Assays using umbilical cord monocyte dendritic cell cultures with the protocol defined in this work, which involves a cytotoxicity study followed by evaluation of several markers of adverse effects on the dendritic cell maturation process, revealed their usefulness for investigating xenobiotic immunotoxicity toward immune primary reactions. PMID- 16703279 TI - Adjuvant treatment of stage II colon cancer: is there a true no-chemotherapy group? PMID- 16703280 TI - Heterologous expression of bacterial and human multidrug resistance proteins protect Escherichia coli against mercury and zinc contamination. AB - In order to determine the role of multidrug resistance proteins in mercury and zinc resistance, human MDR1, Lactococcus lactis lmrA, and Oenococcus oeni omrA genes were expressed in an Escherichia coli tolC mutant which is hypersensitive to metals. The three transporters conferred an increased mercury and zinc resistance to E. coli as compared to the control bacteria. This improved resistance correlated with a decreased zinc and mercury bioaccumulation. Indeed, quantification of intracellular metal concentrations by atomic absorption spectrometry (AAS) showed a 2.1-, 3-, and 5.1-fold decrease in zinc in cells expressing hMDR1, omrA, and lmrA, respectively, and a 2.7-, 7.5-, and 7.7-fold decrease in mercury in cells expressing omrA, lmrA, and hMDR1, respectively, as compared to the control bacteria. This means that hMDR1, LmrA, and OmrA proteins which are specialised in xenobiotic scavenging, their main known function, are nevertheless able to confer some resistance against metals. Our results show that the tolC mutated strain is well adapted to the study of MDR transporter activity and could be used to screen substrates and competitive hMDR1 inhibitors. PMID- 16703281 TI - Reduction of oxidative stress induced vanadium toxicity by complexing with a flavonoid, quercetin: a pragmatic therapeutic approach for diabetes. AB - Vanadium compounds are known to lower blood glucose level in diabetes but are associated with toxicity. In vitro cytotoxicity of VOSO(4) and bis(quercetinato) oxovanadium(IV) (BQOV) was examined in CHO cells. Both the agents showed time and dose dependent increase in ROS generation however it was relatively less in BQOV. Moreover, VOSO(4) also caused higher necrosis. Hypoglycemic potential of VOSO(4) and BQOV was tested in streptozotocin-induced diabetic Balb/c mice. A marked difference was observed in the hypoglycemic action of VOSO(4) and BQOV treated mice that lasted only for about 6 h in VOSO(4) as against 24 h in BQOV. Comparison of acute toxicity of the compounds in normal Balb/c mice revealed negligible nephrotoxicity of BQOV. Kidney analyses of VOSO(4) treated animals' revealed high ROS generation and tubular necrosis. Similarly serum levels of urea and creatinine were elevated in these animals indicating kidney dysfunction. No such abnormality was observed in BQOV treated animals. Reduced nephrotoxicity of BQOV could be due to increased catalase activity found in the kidney of BQOV treated animals and BQOV's radical scavenging activity. The data clearly demonstrates immense hypoglycemic activity and reduced toxicity of BQOV thus making the conjugate a suitable candidate for therapeutic utility. PMID- 16703282 TI - The arterial supply of the temporalis muscle. AB - Owing to the diverse applications of the temporalis muscle in reconstructive surgery, the study of its arterial supply is becoming an issue of great importance nowadays. The material of the present study consisted of 44 specimens, four obtained from two stillbirths and 40 dissected from 20 embalmed cadavers after injecting the external carotid artery with lead oxide solution. Direct branches from the second part of the maxillary artery and the middle temporal artery proved to be constantly furnishing the muscle from its superficial and deep surfaces. The muscular branch of the middle temporal artery supplied the middle and posterior thirds of the superficial surface and the posterior third of the medial surface of the muscle. The superficial temporal artery participated in supplying the muscle from its lateral surface, while the anterior and posterior deep temporal arteries lay deep to the anterior and the middle thirds of the muscle, respectively. The temporal branches of the middle meningeal artery anastomosed with the deep temporal arteries, thereby contributing to the supply of the temporalis muscle. An arterial pedicle arising from the third part of the maxillary artery constituted an additional supply in 9.1% of the specimens, providing an additional arterial pedicle for temporalis-muscle-flap elevation. PMID- 16703283 TI - A case of spontaneous splenorenal shunt associated with splenic artery aneurysm. AB - A case of large spontaneous splenorenal (S-R) shunt accompanied with splenic artery aneurysm is reported in this paper. The large blood vessel, a shunt, anatomizing the splenic vein and left renal vein is macroscopically found during the autopsy of a 71-year-old male, accompanied with liver cirrhosis and splenomegaly. Though there are many reports demonstrating medical imaging, anatomical findings are relatively poor besides describing and illustrating the appearances of the S-R shunt, the splenic artery aneurysm and related branches of the splenic vein and left renal vein, the embryologic and pathogenetic origins of these anomalies are also discussed with the description of the relationship between the S-R shunt and the splenic artery aneurysm. PMID- 16703284 TI - [Evaluation of flat-rate payment in radiation oncology. German experience with disease-related groups for inpatient funding in radiation oncology]. AB - PURPOSE: In Germany a new casemix-related reimbursement system with "diagnosis related groups" (DRGs) for inpatient treatment was started in 2003. The first German system G-DRG 1.0 was developed on the basis of the Australian AR-DRG version 4.1. German inpatient treatment in radiation oncology was not specifically represented in this system due to the very different health care systems. As the DRG system was planned as a pricing system with severe effects on the funding of radiation oncology departments, an adjustment was urgently needed. For the modification, national data about pattern of care and economic relevance were needed. METHODS: For 3,689 cases treated in radiation oncology departments from eleven hospitals data were collected prospectively concerning diagnosis, length of stay, procedures and high-cost drugs and treatments. The DRGs were analyzed for homogeneity in length of stay and costs. Readmission frequency and interval were analyzed and the relevance of existing reimbursement regulations for this situation was evaluated. RESULTS: It could be shown, that radiation therapy implicated additional expenses for oncologic inpatients. These additional costs were not represented in the G-DRG 1.0 reimbursement system. Chemotherapy was an additional cause for economically inhomogeneous oncologic DRGs. The complex sequence of cases for the same patient could be shown, and that the rules for reimbursement of readmissions have to take these sequential treatments into account. Based on these data, modifications of the reimbursement system were suggested. In the following G-DRG version for the year 2004, 21 DRGs were designed for patients receiving radiation therapy. The regulations concerning the readmission of oncologic patients were modified. The correlation between the number of radiation therapy fractions and the total expense was acknowledged in the following year (G-DRG system 2005) and resulted in 35 DRGs. The version for 2006 showed the solidity of these solutions with almost unchanged definitions of these DRGs. CONCLUSION: This evaluation revealed the deficits in the G-DRG system 1.0 (and the AR-DRG system 4.1) related to the inpatient treatment in radiation oncology departments. Modifications could be proposed for following years. In 2004-2006, the regulatory boards adopted several implications of these data for the improvement of the German casemix-based hospital-financing system. PMID- 16703285 TI - Clinical results of intracoronary brachytherapy (ICBT) for multiple in-stent restenosis. AB - BACKGROUND AND PURPOSE: Treatment of in-stent restenosis (ISR) with percutaneous coronary intervention (PCI) alone is often followed by early re-restenosis. The present study focused on the effect of intracoronary brachytherapy (ICBT) on multiple in-stent restenosis (MISR) after repeated PCI. PATIENTS AND METHODS: 40 patients (27 male, 13 female, age: 66 +/- 9 years) with MISR (two to six ISRs, median three ISRs) were retrospectively analyzed. All patients were treated by using the Novoste((R)) Beta-Cathtrade mark 3.5F System after PCI. The target vessel received 18.4-25.3 Gy of radiation at a depth of 2 mm from the center of the source. The restenosis-free survival and overall survival were calculated by Kaplan-Meier analysis (log-rank). The time interval between last PCI without ICBT and the consecutive recurrence was compared with the follow-up time after PCI with ICBT. RESULTS: The 3-year overall survival rate after ICBT was 93%. The 0.5 , 1-, 2-, and 3-year ISR-free survival rates after PCI + ICBT were 81%, 72%, 52%, and 38%, respectively. After PCI alone, the 0.5-, 1-, and 2-year ISR-free survival rates were 30%, 13%, and 0%, respectively. This difference was highly significant (p < 0.0001). Patients with more than three ISRs before ICBT had a better outcome (3-year ISR-free survival: 80%) than patients with only two or three ISRs before ICBT (3-year ISR-free survival: 25%; p < 0.05). CONCLUSION: ICBT is highly effective and safe in patients with ISR. The results of this study are in accordance with the WRIST and BETA-WRIST data. After 6 months both studies revealed an ISR-free survival rate of 86% (WRIST) and 66% (BETA-WRIST), respectively. The ISR rates in the own control group (70%) were comparable to the placebo groups in WRIST (68%) and BETA-WRIST (72%). Interestingly, patients with more than three ISRs before ICBT had the lowest ISR rate after ICBT. PMID- 16703286 TI - Concurrent radiochemotherapy of locally recurrent or advanced sarcomas of the uterus. AB - BACKGROUND: Uterine sarcomas are rare tumors. Until now, no data on the treatment of recurrent or advanced uterine sarcomas using concurrent radiochemotherapy (RCT) has been available. PATIENTS AND METHODS: From 01/1997 to 03/2004, seven patients with locally recurrent (n = 6) or locally advanced uterine sarcomas (n = 1) received concurrent RCT after tumor surgery (R1/2 resection in 3/7 patients). A total radiation dose of 45 Gy was applied in single doses of 1.8 Gy using an external-beam technique; in addition, three to four intracavitary doses of 5 Gy were applied. Concurrent chemotherapy was generally administered as follows: 1.2 g/m(2) ifosfamide on days 1-5 and 29-33 in combination with 50 or 40 mg/m(2) adriamycin on days 2 and 30. 3/7 patients received further cycles of chemotherapy. The median follow- up was 35 months. RESULTS: All recurrences (before RCT) were localized either in the vagina or in or directly proximal to the vaginal stump. The main side effects of RCT were hemotoxicity (grade 3: n = 3/7; grade 4: n = 4/7; neutropenic fever n = 1/7) and diarrhea (grade 3: n = 5/7). At the median follow-up (35 months), 4/7 patients had recurrences (one local recurrence; one lymph node recurrence outside the irradiated field, two distant metastases). Local control in the irradiated field was 80% +/- 18% after 3 years. Disease-free survival calculated according to Kaplan-Meier was 57% +/- 19% after 3 years. Presently, 5/7 patients are still alive, corresponding to a 3 year survival rate of 83% +/- 15%. CONCLUSION: Concurrent RCT shows good local effectiveness with a good long-term survival. Further evaluation in phase II studies is recommended. PMID- 16703287 TI - [Presentation of a 3D conformal radiotherapy technique for head-and-neck tumors resulting in substantial protection of the parotid glands]. AB - PURPOSE: The aim of this study was to improve the irradiation technique for the treatment of head-and-neck tumors and, in particular, to make use of the advantages found in modern 3D planning to protect the parotid glands. PATIENTS AND METHODS: For this investigation the 3D dataset of a standard patient with oropharyngeal carcinoma of UICC stage IVA was used. In the CT scans (slice thickness 5 mm) the planning target volume (PTV), the boost volume and both parotids were delineated. Three different techniques were calculated for two different dose levels (50 Gy for PTV and 64 Gy for boost volume, using single doses of 2 Gy). For technique 1 (T1) a parallel opposed field photon/electron irradiation was designed, for technique 2 (T2) an opposed/arc field irradiation was employed, and for technique 3 (T3) a combination of a static coplanar and arc field irradiation was designed. The sum doses D(min), D(max) and D(mean) for PTV, boost volume, and ipsilateral and contralateral parotid gland were evaluated, and the time needed for calculation of the plans was also determined. RESULTS: For all techniques used, the calculated doses in the PTV (D(min) 5.6 +/- 0.1 Gy, D(max) 73.7 +/- 0.1 Gy, and D(mean) 57.9 +/- 0.5 Gy) and in the boost volume (D(min) 46.9 +/- 1.5 Gy, D(max) 73.8 +/- 0.12 Gy, and D(mean) 65.8 +/- 0.9 Gy) were equal. Significant differences were found regarding the three different techniques, e.g., for the ipsilateral parotid gland D(min) (T1 = 47.4, T2 = 50.6, and T3 = 38.4 Gy) as well as for the contralateral parotid gland D(min) (T1 = 42.1, T2 = 44.2, and T3 = 17.8 Gy) and D(mean) (T1 = 51.3, T2 = 52.8, and T3 = 32.6 Gy). Regarding the three different techniques, significant differences were found in favor of T3. The determined planning times were as follows: T1 = 90, T2 = 60, and T3 = 90 min. CONCLUSION: The combination of static coplanar and arc field technique (T3) resulted in a substantially better protection as compared to both other techniques. This was especially the case with regard to the contralateral parotid gland, when the dose distributions were calculated equally for PTV and boost volume. In this study, the D(mean) dose of the contralateral parotid gland was lower than the TD(50) of 37 Gy (95% confidence interval 32-43 Gy) previously assumed by the authors. Therefore, it can be concluded that in the present study a more intensive protection of this gland and a reduction in xerostomia were possibly obtained. PMID- 16703288 TI - IMRT in hypopharyngeal tumors. AB - BACKGROUND AND PURPOSE: Intensity-modulated radiation therapy (IMRT) data on hypopharyngeal cancer (HC) are scant. In this study, the authors report on early results in an own HC patient cohort treated with IMRT. A more favorable outcome as compared to historical data on conventional radiation techniques was expected. PATIENTS AND METHODS: 29 consecutive HC patients were treated with simultaneous integrated boost (SIB) IMRT between 01/2002 and 07/2005 (mean follow-up 16 months, range 4-44 months). Doses of 60-71 Gy with 2.0-2.2 Gy/fraction were applied. 26/29 patients were definitively irradiated, 86% received simultaneous cisplatin-based chemotherapy. 60% presented with locally advanced disease (T3/4 Nx, Tx N2c/3). Mean primary tumor volume measured 36.2 cm(3) (4-170 cm(3)), mean nodal volume 16.6 cm(3) (0-97 cm(3)). RESULTS: 2-year actuarial local, nodal, distant control, and overall disease-free survival were 90%, 93%, 93%, and 90%, respectively. In 2/4 patients with persistent disease (nodal in one, primary in three), salvage surgery was performed. The mean dose to the spinal cord (extension of > 5-15 mm) was 26 Gy (12-38 Gy); the mean maximum (point) dose was 44.4 Gy (26-58.9 Gy). One grade (G) 3 dysphagia and two G4 reactions (laryngeal fibrosis, dysphagia), both following the schedule with 2.2 Gy per fraction, have been observed so far. Larynx preservation was achieved in 25/26 of the definitively irradiated patients (one underwent a salvage laryngectomy); 23 had no or minimal dysphagia (G0-1). CONCLUSION: Excellent early disease control and high patient satisfaction with swallowing function in HC following SIB IMRT were observed; these results need to be confirmed based on a longer follow-up period. In order to avoid G4 reactions, SIB doses of < 2.2 Gy/fraction are recommended for large tumors involving laryngeal structures. PMID- 16703289 TI - Quantifying radioxerostomia: salivary flow rate, examiner's score, and quality of life questionnaire. AB - BACKGROUND AND PURPOSE: Salivary flow rates alone are not sufficient to quantify all aspects of radioxerostomia. This is a problem in studies aiming to reduce radioxerostomia. The aim of this study was to evaluate the association between objectively measured salivary flow rate and subjective xerostomia ratings by the physician (RTOG scale) or the patients (quality of life [QoL] questionnaire). PATIENTS AND METHODS: In a case-control study patients who underwent recall for oral cancer were screened. Inclusion criteria for this diagnostic, noninterventional study were: history of oral carcinoma, surgical and radiation therapy, time interval from start of radiation therapy > 90 days, salivary glands within the radiation field. The control group consisted of patients, who had not received radiotherapy. RTOG salivary gland score, quality of life (EORTC QLQ-C30 and H&N35), and sialometry were recorded. RESULTS: Patients with RTOG score 0 had mean salivary flow rates of 0.3 ml/min, those with RTOG 1 0.12 ml/min, RTOG 2 0.02 ml/min, and RTOG 3 < 0.01 ml/min. RTOG score 4 (total fibrosis) did not occur. Based on salivary flow rates, all patients were grouped into xerostomia < 0.2 ml/min (30 patients) and nonxerostomia (twelve patients). QoL results revealed significant differences between patients with xerostomia and nonxerostomia for physical function, dyspnea, swallowing, social eating, dry mouth, nutritional support, and a tendency to higher values for appetite loss. CONCLUSION: The correlation between "subjective" QoL parameters and salivary flow was confirmed. The different subjective aspects of radioxerostomia seem to be better differentiated by the EORTC QoL questionnaire. PMID- 16703290 TI - A dosimetric comparison of IORT techniques in limited-stage breast cancer. AB - BACKGROUND AND PURPOSE: For intraoperative radiotherapy (IORT) during breast conserving treatment four different techniques have been addressed: interstitial brachytherapy, an inflatable balloon with a central high-dose-rate source (MammoSite), a miniature orthovolt system (Intrabeam), and linac-based electron radiotherapy (IOERT). The dosimetric properties of these methods are compared. MATERIAL AND METHODS: Planning target volumes (PTVs) of the same size but of different shapes are assumed, corresponding to the technique's specific situs. Dose distributions for the PTVs and for surrounding tissues are demonstrated by dose-volume histograms and a list of physical parameters. A dose inhomogeneity index (DII) is introduced to describe the deviation of a delivered from the prescribed dose, reaching its minimal value 0 in case of perfect homogeneity. RESULTS: In terms of DII, IOERT reaches the lowest value followed by the MammoSite, the Intrabeam and interstitial implants. The surrounding tissues receive the smallest average dose with IOERT, closely followed by the orthovolt system. CONCLUSION: When comparing simplified geometric figures, IOERT delivers the most homogeneous dose distributions. However, in clinical reality PTVs often present asymmetric shapes instead of ideal geometries. Due to a strictly centric dose fall-off, any system with a round central applicator will have technical limits. During IOERT margin-directed applicator guidance is possible and interstitial brachytherapy allows for polygonal dose shaping. These techniques seem to be superior for asymmetric PTV irradiation. PMID- 16703291 TI - A process for making cutaneous radiation applicators based on digital data. AB - An optical modeling process for facial regions and other body surfaces has been developed. The body part in question is digitized using optical 3-D metrology to obtain a comprehensive dataset. The data is then prepared for further use by smoothing the point clusters. The radiophysically significant position of the radioactive material, e. g., seeds, aluminum tubes, etc., can be accurately determined using CAD modeling. Subsequently, a rapid prototyping process (fused deposition modeling [FDM]) is used to implement the CAD model directly in order to create a radiation applicator that can be used in practice. Biologically compatible polycarbonate can be used for this purpose. PMID- 16703292 TI - What's the best way to measure antibiotic use in hospitals? PMID- 16703293 TI - Evaluation of safety and efficacy of rosiglitazone in the treatment of HIV associated lipodystrophy syndrome. AB - BACKGROUND: HIV-associated lipodystrophy syndrome (LDS) as a long-term side effect of HAART is becoming increasingly important and negatively affects adherence to medication. Currently, an effective therapy is not available. There is some evidence that the drug class of thiazolidindiones might be effective in the treatment of LDS. PATIENTS AND METHODS: Prospective open-label study with 20 HIV-infected patients suffering from severe LDS. Patients received 4 mg rosiglitazone once daily for a 24-week study period. Efficacy was assessed by measurement of metabolic and anthropometric parameters, total body DXA scan, CT scan of the abdomen, photo documentation and self-assessment. RESULTS: Rosiglitazone treatment was well tolerated. DXA scans demonstrated a highly significant increase in adipose tissue of the limbs (2644 +/- 1334 g vs 3380 +/- 1614 g, p < or = 0.001) without any change in total fat mass. Abdominal CT-scans revealed a significant increase in subcutaneous adipose tissue (113.7 +/- 82.4 cm(2) vs 125.3 +/- 83.7 cm(2), p = 0.04). Abdominal circumference decreased significantly (94.7 +/- 8.7 cm vs 92.2 +/- 8.45 cm, p = 0.03) without any relevant change of body weight or BMI. We observed an increase in serum cholesterol (248 vs 281 mg/dl, p = 0.006) and serum triglycerides (301 vs 351 mg/dl, p = 0.1). Furthermore, no side effects of clinical relevance were observed. The insulin sensitivity index improved without reaching statistical significance. Thirteen patients (65%) reported general improvement of LDS symptoms. Evaluation of photo documentation by five HIV-experts revealed poor concordance and no relevant change of LDS. CONCLUSIONS: The results of this study suggest that rosiglitazone is safe in the treatment of HAART-associated lipodystrophy and has moderate clinical efficacy. We found a trend towards improved insulin sensitivity and as a possible limiting factor an unfavorable increase in serum cholesterol and triglycerides. PMID- 16703294 TI - Evolution of hepatitis C virus quasispecies in children with chronic hepatitis C. AB - BACKGROUND: Hepatitis C virus (HCV) circulates as a mixture of different but closely related genomes: this quasispecies nature could be essential for virus persistence and could induce resistance to interferon therapy. Since little is known on the behavior of HCV quasispecies in children and adolescents with chronic hepatitis C, we analyzed the virus population in six untreated children during a 5-year follow-up. METHODS: Six children aged 1-8 years, infected early in life with HCV, were included in the study. From each of them, 2 or 3 sequential serum samples obtained over a 5-year follow-up period were examined. The HCV quasispecies heterogeneity and diversity in the E2 hypervariable region-1 (HVR-1) were analyzed among samples by the heteroduplex mobility assay, and the distance between variants was estimated by the heteroduplex mobility ratio (HMR). RESULTS: The HCV population was initially highly homogeneous in all six children. During follow-up, diversification of HVR-1 leading to a more complex viral population occurred in all cases, and was particularly evident in the three older children (HMR: 0.82-0.54). Changes in the HVR-1 sequence occurred without relation to the profile of ALT and HCV-RNA levels. CONCLUSIONS: HCV quasispecies diversification is a common event during chronic hepatitis C in childhood. Host and environmental pressure could be major determinants. The increasing viral heterogeneity could impair the response to antiviral therapy, thus indicating a rationale for early antiviral treatment in children with chronic hepatitis C. PMID- 16703295 TI - Assessment of the clinical significance of production of extended-spectrum beta lactamases (ESBL) by Enterobacteriaceae. AB - BACKGROUND: We conducted a retrospective, cohort-controlled study to evaluate the effect of extended-spectrum beta-lactamase (ESBL) production by Enterobacteriaceae isolated from blood cultures, and of third or fourth generation cephalosporin treatment, on outcome. METHODS: Four hundred and fifty patient-unique Enterobacteriaceae, isolated from blood cultures during 2000 (before routine ESBL testing was introduced), were tested for ESBL by double-disk method and by E-test, assessing cefotaxime, ceftazidime and cefpodoxime, with and without clavulanate. Cases consisted of ESBL-positive (+) samples, originally reported as ceftazidime-susceptible; controls were ESBL-negative (-). Patient records were extensively reviewed. RESULTS: We identified 68 Enterobacteriaceae that were ESBL(+); they were compared with 186 ESBL(-) control organisms. Patients with sepsis due to an ESBL(+) organism more often had nosocomial infection, resided in nursing homes, were functionally dependent, had an indwelling catheter, had Klebsiella, and had a lower serum albumin level (all p < 0.001). Survival of patients with ESBL(+) and ESBL(-) sepsis was, respectively, 71% and 84% (p < 0.05). Multivariate analysis revealed that the only independent risk factor for death was a low serum albumin. Neither empiric nor definite treatment with cephalosporins was found to be an independent risk factor for death. Subset analysis was conducted on 15 patients with ESBL(+) sepsis and 21 controls with ESBL(-) sepsis, who had been treated with ceftazidime or cefepime only. In this subset, ESBL(+) patients more often resided in nursing homes (< 0.05), they had a significantly lower APACHE-II score (< 0.01) and the infection was more often nosocomial (< 0.005). Survival of ESBL(+) and ESBL(-) patients was 67% and 71%, respectively (NS). Time till defervescence did not differ between cases and controls. CONCLUSION: Mortality of patients with ESBL(+) sepsis was higher than that of patients with ESBL(-) sepsis. The reason appears to be related to other factors rather than to empiric treatment with cephalosporins or the nature or resistance pattern of the organism. This, at least, appears to be the case for patients with urosepsis, who constituted the majority of patients in this study. PMID- 16703296 TI - Fever of unknown origin in Taiwan. AB - BACKGROUND: Fever of unknown origin (FUO) is a challenging problem worldwide. There was no prospective study of FUO in the past two decades in Taiwan. A prospective study was conducted. MATERIALS AND METHODS: The prospective study was undertaken from March 2001 to May 2002. All patients fulfilling the modified criteria for FUO, either admitted, referred or consulted in a medical center in southern Taiwan, were enrolled for analysis. RESULTS: A total of 94 cases met the criteria of FUO. The final diagnoses of FUO consisted of 54 infectious diseases (57.4%), 8 hematologic/neoplastic (8.5%), 7 noninfectious inflammatory (7.4%), 8 miscellaneous (8.5%) and 17 undiagnosed (18.1%) cases. The single most common cause of FUO was tuberculosis. Some infectious diseases, such as rickettsiosis and melioidosis, were rarely reported in western countries. Three patients with hemophagocytotic syndrome without ascertainable etiologies were present with FUO in this study. Between the patients with and those without a final diagnosis, the short-term survival (3 months) was compared by the Kaplan-Meier analysis, which revealed no difference. CONCLUSIONS: Mycobacteriosis is still the leading cause of FUO in Taiwan and it is important to identify this treatable disease from all causes of FUO. This study has showed geographical variation among the studies of FUO. PMID- 16703297 TI - In vitro inhibition of coagulase-negative staphylococci by vancomycin/aminoglycoside-loaded cement spacers. AB - BACKGROUND: Successful treatment of allograft infections by the temporary implantation of an antibiotic-loaded polymethylmethacrylate cement spacer depends on the diffusion of antibiotics out of the cement and inhibition of bacterial growth in the surrounding tissue. We investigated with an in vitro model how long antibiotics are released by the cement and if gentamicin-resistant coagulase negative staphylococci (CNS) are inhibited by vancomycin mixed with the gentamicin-loaded cement. MATERIALS AND METHODS: Four formulations of antibiotic loaded cement disks, i.e. gentamicin, tobramycin, vancomycin and tobramycin combined with vancomycin, respectively, were used to test the inhibition of eight isolates of Staphylococcus epidermidis and two reference strains of Staphylococcus aureus by an agar diffusion test on Mueller-Hinton (MH) agar similar to the routine laboratory disk diffusion method. Moreover, cement spacer cylinders loaded with gentamicin alone or combined with vancomycin were submerged in MH agar for weeks and the capacity to inhibit five different isolates of S. epidermidis was measured. RESULTS: The size of the inhibition zones around the antibiotic-loaded cement disks correlated with the minimal inhibitory concentration (MIC) of the antibiotics against the tested strains. All five strains of S. epidermidis were inhibited by vancomycin-loaded cement spacers for at least 30 days. However, two gentamicin-resistant S. epidermidis strains with MICs of 4 mg/l and 16 mg/l could not be inhibited longer than 3 days by the gentamicin-loaded cement spacer. CONCLUSION: The in vitro data suggest that antibiotic-loaded cement spacers inhibit susceptible bacteria for 4-6 weeks. The addition of vancomycin to commercial aminoglycoside-loaded cements might be helpful in allograft infections in tumor patients to inhibit a broad range of bacteria including gentamicin-resistant CNS very commonly found in such infections. PMID- 16703298 TI - Primary cytomegalovirus infection in otherwise healthy adults with Fever of unknown origin: a 3-year prospective survey. AB - BACKGROUND: Primary cytomegalovirus disease is probably still underestimated or missed in common clinical practice, and further prevalence studies should be performed, in particular in the setting of fever of under-determined origin (FUO) in adults. PATIENTS AND METHODS: In a 3-year prospective survey of 123 consecutive adult patients referred for FUO often associated with a broad spectrum of constitutional signs and symptoms, 18 patients (14.6%) were found to have a primary cytomegalovirus infection, after a clinical, instrumental and laboratory workup. RESULTS: In the majority of cases, this syndrome was consistently associated with altered white blood cell count, abnormal T lymphocyte subsets and ultrasonography-confirmed hepatosplenomegaly. On the other hand, altered white blood cell differential and serum hepatic enzymes, and constitutional signs and symptoms were absent in 11.1-27.8% of cases, and an initial laboratory cross-reaction with anti-Epstein-Barr IgM antibodies was detected in 44.4% of episodes. Non-specific signs and symptoms were the only features in 27.8% of patients with adult cytomegalovirus disease, thus, confirming that this disorder may be still clinically underestimated, until virologic assays are performed. A prolonged and varied spectrum of subjective disturbances (similar to those encountered in infectious mononucleosis), which often limited daily activities, involved nearly 30% of subjects, and lasted for 3 15 months after recovery of acute cytomegalovirus disease. CONCLUSION: In the clinical, laboratory, and instrumental workup for FUO, rapid recognition of a primary cytomegalovirus disease is useful to exclude alternative diagnoses, avoid non-necessary exposure to antibiotics, and reassure patients of their self limiting, benign disorder. PMID- 16703299 TI - Trends in antibiotic use at a university hospital: defined or prescribed daily doses? Patient days or admissions as denominator? AB - BACKGROUND: Hospital antibiotic use can be measured by calculating daily doses as defined by the WHO/ATC index (DDD) divided by the number of patient or occupied bed days. We wondered whether changes in antibiotic use density over time at a university hospital using this data format are similar in order of magnitude when compared with a different, alternative dose definition and the number of admissions rather than the number of patient days as denominator. METHODS: Data obtained from the hospital pharmacy for the medical and surgical services of a 1,000-bed university hospital for the period 1992 through 2003 were expressed both in daily doses per 100 patient days and daily doses per admission. A PDD dose definition (prescribed daily doses), defining doses that reflect the usually prescribed dose in adult hospitalized patients with normal renal function was compared with the WHO/ATC 2001 DDD dose definitions. The percent changes using the different data formats between two 3-year averages (1992-1994 and 2001-2003) were calculated. RESULTS AND CONCLUSION: The DDD/100 patient days data format overestimated antibiotic use density changes in this hospital both in medicine (81% vs 48%) as well as in surgery (69% vs 39%) when compared with PDD/100 patient days. Due to changes in the number of admissions and length of stay over the years, the percent change between the two periods expressed in doses per 100 patient days in addition differed substantially from that estimated by using the DDD per admission or PDD per admission data format. Studies evaluating the evolution of hospital antibiotic use need to address the limitations and adequacy of the different data formats. PMID- 16703300 TI - Evaluation of S. aureus ID, a chromogenic agar medium for the detection of Staphylococcus aureus. AB - BACKGROUND: Screening for staphylococci among various patient populations has become important for appropriate therapeutic management and for control of nosocomial infections. The purpose of this study is to evaluate the in vitro sensitivity and specificity of a chromogenic agar medium, S. aureus ID (bioMerieux, France), for the identification of Staphylococcus aureus. MATERIALS AND METHODS: A well-defined collection of S. aureus and coagulase-negative staphylococci (CNS) was used. The methicillin-resistant S. aureus (MRSA) isolates were collected in The Netherlands and all had a unique typing pattern. The methicillin-susceptible S. aureus (MSSA) and CNS were isolated from cultures of blood. The isolates were inoculated on Columbia agar plates with 5% sheep blood and incubated for 24 h at 35 degrees C. From the resulting cultures, a suspension of 0.5 McFarland was made and subsequently 10 mul was streaked on a S. aureus ID plate using a sterile loop. The results were read after 24 h and 48 h of incubation at 35 degrees C. Growth of colonies showing green coloration was considered to be positive (indicating S. aureus). RESULTS: A total of 519 S. aureus strains were tested (249 MSSA, 270 MRSA). The sensitivity to detect S. aureus was 96.5% (501/519) after 24 h and 97.5% (506/519) after 48 h. A total of 478 CNS were tested. The specificity was 98.5% (471/478) after 24 h and 98.3% (470/478) after 48 h. The differences between 24 h and 48 h incubation were not statistically significant. CONCLUSION: S. aureus ID is highly sensitive and specific to differentiate between S. aureus and CNS in vitro. Since the performance does not significantly differ between 24 h or 48 h of incubation, samples need only 1 day of incubation before optimal results can be obtained. PMID- 16703301 TI - Necrotizing gastritis due to Bacillus cereus in an immunocompromised patient. AB - Bacillus cereus is increasingly being acknowledged as a serious bacterial pathogen in immunocompromised patients. We present a case of acute necrotizing gastritis caused by B. cereus in a 37-year-old woman with acute myeloblastic leukemia, who recovered following total parenteral nutrition and treatment with imipenem and vancomycin. B. cereus was isolated from gastric mucosa and blood cultures. Up to now, no case of acute necrotizing gastritis due to this organism has been reported. PMID- 16703302 TI - Neuroborreliosis in an HIV-1 positive patient. AB - Simultaneous co-infections of Borrelia burgdorferi sensu lato and HIV-1 are rare events, with only six published cases. A case of acute neuroborreliosis with facial palsy, meningoradiculitis (Bannwarth's syndrome) in an HIV-1 positive individual is described. Diagnosis was confirmed by Western immunoblot analysis of serum and CSF and by proof of intrathecal production of antibodies against B. garinii. The patient was successfully treated with cefotaxime. In all published HIV+ cases, the course of borreliosis did not differ from that of the HIV negative population and the prognosis in properly treated patients was good. PMID- 16703303 TI - Central venous catheter infection with Brevibacterium sp. in an immunocompetent woman: case report and review of the literature. AB - Brevibacterium spp. were considered apathogenic until a few reports of infections in immunocompromised patients were published. Herein, we present a case of a catheter-related septicemia with Brevibacterium casei in an immunocompetent patient receiving continuous iloprost infusion for pulmonary arterial hypertension and review the clinical presentation of this mainly emerging opportunistic pathogen. PMID- 16703304 TI - Seroprevalence of antibodies to Anaplasma phagocytophilum in Antalya, Turkey. PMID- 16703305 TI - Serious doubts on safety and efficacy of CCR5 antagonists : CCR5 antagonists teeter on a knife-edge. PMID- 16703306 TI - MR imaging of the uterus and cervix in healthy women: determination of normal values. AB - The purpose of this study was to establish normal values for the volume of the uterus and cervix in MRI based on age and the menstrual cycle phase. We performed MRI of the pelvis in 100 healthy women. For the uterus, they were further divided into two groups: one with myomas and/or adenomyosis and one without either. The volume of the uterus and cervix and thickness of the uterine wall layers were analysed by age and the menstrual cycle phase. The mean volume of the uterus in both groups and the cervix significantly increased with age to reach its peak at 41-50 years, and then dropped. Likewise, the thickness of the endometrium and the junctional zone, but not the myometrium, significantly increased until 41-50 years, and then decreased. When we compared the volume of the uterus and cervix and the thickness of the uterine wall layers between the two phases of the menstrual cycle, we found no significant differences. The volume of the uterus and cervix and the thickness of the endometrium and junctional zone differ significantly with age, but not between the two phases of the menstrual cycle. Knowledge of MRI-related normal values can be expected to aid the early identification of uterine pathologies. PMID- 16703307 TI - Prospective intraindividual comparison between respiratory-triggered balanced steady-state free precession and breath-hold gradient-echo and time-of-flight magnetic resonance imaging for assessment of portal and hepatic veins. AB - The purpose of this study was to compare respiratory-triggered balanced steady state free precession (bSSFP) with breath-hold contrast-enhanced dynamic two dimensional (2D) gradient-echo (GRE) and time-of-flight (TOF) magnetic resonance imaging (MRI) for portal and hepatic vein visualization and assessment of portal and hepatic venous variants. Sixty patients with liver disease underwent nonenhanced bSSFP and contrast-enhanced GRE, bSSFP, and TOF imaging. Contrast-to noise ratios (CNRs) for portal and hepatic veins were measured. Two readers rated the quality of portal and hepatic vein visualization on a 5-point Likert scale. The diagnostic performance of each MRI series in the detection of portal and hepatic venous variants was assessed in 40/60 patients who also underwent contrast-enhanced multidetector-row computed tomography (MDCT). CNRs for portal and hepatic veins were highest on contrast-enhanced bSSFP images. Image quality of portal and hepatic veins was rated higher for nonenhanced bSSFP than for contrast-enhanced GRE (p<0.03) and TOF (p<0.003) and higher for contrast-enhanced than for nonenhanced bSSFP (p<0.003). Compared with MDCT, portal and hepatic venous variants were identified with an accuracy of 99% on bSSFP images, with an excellent interobserver agreement (kappa=0.97). Compared with MDCT, presence of surgically important portal and hepatic venous anatomical variants can be predicted with high accuracy on bSSFP images. PMID- 16703308 TI - Reader error during CT colonography: causes and implications for training. AB - This study investigated the variability in baseline computed tomography colonography (CTC) performance using untrained readers by documenting sources of error to guide future training requirements. Twenty CTC endoscopically validated data sets containing 32 polyps were consensus read by three unblinded radiologists experienced in CTC, creating a reference standard. Six readers without prior CTC training [four residents and two board-certified subspecialty gastrointestinal (GI) radiologists] read the 20 cases. Readers drew a region of interest (ROI) around every area they considered a potential colonic lesion, even if subsequently dismissed, before creating a final report. Using this final report, reader ROIs were classified as true positive detections, true negatives correctly dismissed, true detections incorrectly dismissed (i.e., classification error), or perceptual errors. Detection of polyps 1-5 mm, 6-9 mm, and > or =10 mm ranged from 7.1% to 28.6%, 16.7% to 41.7%, and 16.7% to 83.3%, respectively. There was no significant difference between polyp detection or false positives for the GI radiologists compared with residents (p=0.67, p=0.4 respectively). Most missed polyps were due to failure of detection rather than characterization (range 82-95%). Untrained reader performance is variable but generally poor. Most missed polyps are due perceptual error rather than characterization, suggesting basic training should focus heavily on lesion detection. PMID- 16703309 TI - In vivo proton MR spectroscopy of primary tumours, nodal and recurrent disease of the extracranial head and neck. AB - Benign and malignant neoplasms as well as metastatic lymph nodes of 39 patients were examined using localized single voxel magnetic resonance spectroscopy (MRS) [repetition time (TR) 1500, echo time (TE) 135) at 1.5 T. New techniques with simultaneous correction of motion artefacts during the acquisition, three dimensional saturation pulses, respiratory triggering and smaller volume of interest (VOI) size, were applied. Ratios of peak areas under the choline (Cho) and creatine (Cr) resonances were estimated in all cases and compared with those from samples of normal tissue. Ninety one spectra were acquired in 39 patients, 63 of which were suitable for further evaluation. The smallest VOI was 0.40 cm(3). The Cho/Cr ratios in all malignant neoplasms (mean: 5.2, range: 1.7-17.8) were significantly elevated relative to those in the normal muscle structures (mean: 0.9, range: 0.2-1.4), while those in the benign neoplasms were elevated (mean: 24.4, range: 1.4-59.7) with respect to those in the malignant ones. The average Cho/Cr ratio in the metastatic lymph nodes was significantly higher (mean: 4.8, range: 3.3-5.6) than that for benign lymphoid hyperplasia (mean: 2.2, range: 1.0-3.0). MRS measurements were able to differentiate recurrent disease from post-therapeutic tissue changes in 11 out of 13 patients. PMID- 16703310 TI - Diagnostics and characterisation of preocclusive stenoses and occlusions of the internal carotid artery with B-flow. AB - The purpose was to evaluate whether B-flow can improve the ultrasonographic diagnosis of preocclusive stenosis and occlusion of the internal carotid artery (ICA) compared with colour-coded Doppler and power Doppler. Ninety patients with occlusions or preocclusive stenoses of the ICA suspected by Doppler sonography were examined with B-flow in comparison with colour-coded Doppler sonography (CCDS), power Doppler (PD) and intra-arterial digital subtraction angiography (DSA). Intrastenotic flow detection and lengths of stenoses were the main criteria. Ulcerated plaques found by surgery in 42/90 patients were compared by ultrasonography (US). Diagnosis of ICA occlusion with CCDS, PD and B-flow was correct in all 42 cases. A preocclusive ICA stenosis in DSA was detected correctly in all 48/48 cases (100%) for B-flow, in 44/48 (92%) for PD and in 39/48 (81%) for CCDS. Surgical findings showed in 17/42 cases ulcerated plaques; 15/17 (89%) of these cases were detected with B-flow, 12/17 (71%) with PD, 10/17 (59%) with CCDS, and 8/17 (47%) with DSA. With B-flow the extent of stenosis was appraised more precisely than with PD and CCDS (P<0.0001). In conclusion, B-flow is a reliable method for preocclusive stenosis of the ICA with less intrastenotic flow artefacts. B-flow facilitates the characterization of plaque morphologies. PMID- 16703311 TI - Image quality in CT perfusion imaging of the brain. The role of iodine concentration. AB - The purpose of this study was to evaluate the impact of various iodine contrast concentrations on image quality in computed tomography (CT) perfusion studies. Twenty-one patients with suspicion of cerebral ischemia underwent perfusion CT using two different iodine contrast concentrations: 11 patients received iomeprol 300 (iodine concentration: 300 mg/ml ) while ten received the same volume of iomeprol 400 (iodine concentration: 400 mg/ml). Scan parameters were kept constant for both groups. Maps of cerebral blood flow (CBF), cerebral blood volume (CBV), and time to peak (TTP) were calculated from two adjacent slices. Quantitative comparisons were based on measurements of the maximum enhancement [Hounsfield units (HU)] and signal-to-noise index (SNI) on CBF, CBV, and TTP images. Determinations of grey-to-white-matter delineation for each iodine concentration were performed by two blinded readers. Only data from the non ischemic hemispheres were considered. Both maximum enhancement and SNI values were higher after iomeprol 400, resulting in significantly better image quality in areas of low perfusion. No noteworthy differences were found for normal values of CBF, CBV, and TTP. Qualitative assessment of grey/white matter contrast on CBF and CBV maps revealed better performance for iomeprol 400. For brain perfusion studies, highly concentrated contrast media such as iomeprol 400 is superior to iomeprol 300. PMID- 16703312 TI - Percutaneous radiofrequency ablation of lung tumors with expandable needle electrodes: current status. AB - Percutaneous radiofrequency thermal ablation (RFA) has been used to treat primary and secondary liver tumors under ultrasound, computed tomography (CT), or magnetic resonance imaging (MRI) guidance for the past decade [Park et al., Radiol Clin North Am 38:545-561, 2000; Siperstein and Gotomirski, Cancer J 6:S293 S301, 2000; Kelekis et al., Eur Radiol 13:1100-1105, 2003]. RFA is a low-cost, minimally invasive treatment that has recently attracted attention for treating tumors in different solid organs with promising results [Dupuy and Goldberg, J Vasc Interv Radiol 12:1135-1148, 2001; Friedman et al., Cardiovasc Intervent Radiol 27:427-434, 2004]. It can be provided with minimal hospitalization, and experienced practitioners have reported low complication rates [Dupuy and Goldberg, J Vasc Interv Radiol 12:1135-1148, 2001; Livraghi et al., Radiology 226:441-451, 2003]. Patients with lung malignancies (primary lung cancer or pulmonary metastases), who cannot be operated, might be candidates for RFA treatment. It can also be used in association with other treatments (i.e., chemotherapy, radiotherapy) for better disease control. Combination of the above with RFA may help reduce morbidity and mortality. Many ways to apply energy to the tumor exist (monopolar and bipolar RFA, microwave, laser, brachytherapy). In this review we will focus on expandable monopolar systems, which despite their deficiencies are the most popular in the interventional radiology sector. PMID- 16703313 TI - Amyloidosis of the heart and respiratory system. AB - Since the incidence of amyloidosis is increasing, the purpose of this article is to review the imaging features of intrathoracic amyloidosis. Amyloidosis forms a heterogeneous group of disorders characterised by the extracellular deposition of a homologous protein complex. The heart is the most commonly involved organ in the chest. Respiratory amyloidal deposition is much less common and may be generalised, when it occurs as a part of a systemic disease, or it may be restricted only to the respiratory system. Although, the abnormalities are considered non-specific, recent literature suggests-especially for cardiac amyloidosis-specific patterns of abnormalities. PMID- 16703314 TI - MR liver imaging and cholangiography in the presence of surgical metallic clips at 1.5 and 3 Tesla. AB - To evaluate whether clips from prior cholecystectomy impair image quality during magnetic resonance cholangiography (MRC) at 3 Tesla (T) compared with 1.5 T, surgical clips were embedded in a gel phantom and positioned at predefined distances from a fluid-filled tube designed to simulate the bile duct. The maximum clip distance was noted where susceptibility artifacts obscured the fluid filled tube at 1.5 T and 3 T. Susceptibility artifact size was calculated for each sequence within each magnet class. In vivo analysis included 42 patients postcholecystectomy who underwent MRC at either 1.5 T or 3 T. In vitro, mean area of susceptibility artifacts was 104 mm2 on 3-T and 75 mm2 on 1.5-T MR imaging (MRI). While surgical clips within a 2-mm range impaired visualization of the fluid-filled tube on 1.5-T MRI, this range increased to 4 mm on 3-T MRI. In vivo, MRC image quality was impaired by susceptibility artifacts in three of 21 cases at 3 T and in two of 21 cases at 1.5 T. Overall, biliary pseudo-obstructions due to susceptibility artifacts from cholecystectomy surgical clips were not substantially more common on 3-T MRC in clinical practice, and patients with a history of prior cholecystectomy should not be excluded from a 3-T MRC. PMID- 16703315 TI - Letter to the editor concerning "HPV in anal squamous cell carcinoma and anal intraepithelial neoplasia (AIN)" by Varnai et al. (Int J Colorectal Dis 21:135 142, 2006). PMID- 16703316 TI - Tailgut cysts: diagnostic challenge for both pathologists and clinicians. PMID- 16703317 TI - Perianal Langerhans cell histiocytosis. PMID- 16703321 TI - Functional expression of Candida antarctica lipase B in the Escherichia coli cytoplasm--a screening system for a frequently used biocatalyst. AB - In this paper, we report for the first time the functional expression of lipase B from the yeast Candida antarctica (CalB) in the Escherichia coli cytoplasm. The enzyme possessing three disulfide bonds was functionally expressed in the strain Origami B. Expression under the control of a lac promoter yielded 2 U mg(-1), whereas expression of a thioredoxin-CalB fusion protein yielded 17 U mg(-1). The native enzyme was most efficiently expressed under control of the cspA promoter (11 U mg(-1)). Coexpression of different chaperones led to a strong increase in active protein formation (up to 61 U mg(-1)). A codon-optimized synthetic variant of calb did not show significant effects on functional protein yield. Functional CalB expression was not only achieved in shake flasks but also in microtiter plate scale. Therefore, this CalB expression system is suitable for high throughput applications, including the screening of large gene libraries as those derived from directed evolution experiments. PMID- 16703320 TI - Expression of the sweet-tasting plant protein brazzein in Escherichia coli and Lactococcus lactis: a path toward sweet lactic acid bacteria. AB - Brazzein is an intensely sweet-tasting plant protein with good stability, which makes it an attractive alternative to sucrose. A brazzein gene has been designed, synthesized, and expressed in Escherichia coli at 30 degrees C to yield brazzein in a soluble form and in considerable quantity. Antibodies have been produced using brazzein fused to His-tag. Brazzein without the tag was sweet and resembled closely the taste of its native counterpart. The brazzein gene was also expressed in Lactococcus lactis, using a nisin-controlled expression system, to produce sweet-tasting lactic acid bacteria. The low level of expression was detected with anti-brazzein antibodies. Secretion of brazzein into the medium has not led to significant yield increase. Surprisingly, optimizing the codon usage for Lactococcus lactis led to a decrease in the yield of brazzein. PMID- 16703322 TI - The role of PHB metabolism in the symbiosis of rhizobia with legumes. AB - The carbon storage polymer poly-beta-hydroxybutyrate (PHB) is a potential biodegradable alternative to plastics, which plays a key role in the cellular metabolism of many bacterial species. Most species of rhizobia synthesize PHB but not all species accumulate it during symbiosis with legumes; the reason for this remains unclear, although it was recently shown that a metabolic mutant of a nonaccumulating species retains the capacity to store PHB in symbiosis. Although the precise roles of PHB metabolism in these bacteria during infection, nodulation, and nitrogen fixation are not determined, the elucidation of these roles will influence our understanding of the metabolic nature of the symbiotic relationship. This review explores the progress that was made in determining the biochemistry and genetics of PHB metabolism. This includes the elucidation of the PHB cycle, variations in PHB metabolism among rhizobial species, and the implications of these variations, while proposing a model for the role of PHB metabolism and storage in symbiosis. PMID- 16703323 TI - Inhibitors of kinesin Eg5: antiproliferative activity of monastrol analogues against human glioblastoma cells. AB - The inhibition of kinesin Eg5 by small molecules such as monastrol is currently evaluated as an approach to develop a novel class of antiproliferative drugs for the treatment of malignant tumours. Therefore, we studied the effects of the new monastrol analogues enastron, dimethylenastron and vasastrol VS-83 on the proliferation of human glioblastoma cells in the kinetic crystal violet assay. Compared to monastrol, the new cell cycle specific compounds showed an at least one order of magnitude higher anti proliferative activity against U-87 MG, U-118 MG, and U-373 MG glioblastoma cells. The compounds were neither inactivated by hydrolysis nor by binding to serum proteins. Moreover, we demonstrated the characteristic monoaster formation after incubation of cells with the new compounds by confocal laser scanning microscopy. We also showed that the arrangement of beta-actin and tubulin, vital components of the cyto-skeleton of mitotic and quiescent cells, were not affected by the new compounds. Due to the necessity of overcoming the blood-brain barrier in the treatment of brain tumours, we investigated if the new monastrol analogues are modulators or substrates of the p-glycoprotein (p-gp) 170 by a flow cytometric calcein-AM efflux assay. The tested compounds showed no modulating effects on the p-gp function. With respect to the treatment of primary and secondary CNS tumours, the results of our experiments suggest that the new monastrol analogues represent an interesting class of potential anticancer drugs, predicted to be less neurotoxic in comparison to classical tubulin inhibitors. PMID- 16703326 TI - Effects of arginine treatment on nutrition, growth and urea cycle function in seven Japanese boys with late-onset ornithine transcarbamylase deficiency. AB - BACKGROUND: The aim of this study was to investigate the effects of arginine on nutrition, growth and urea cycle function in boys with late-onset ornithine transcarbamylase deficiency (OTCD). Seven Japanese boys with late-onset OTCD enrolled in this study resumed arginine treatment after the cessation of this therapy for a few years. Clinical presentations such as vomiting and unconsciousness, plasma amino acids and urinary orotate excretion were followed chronologically to evaluate urea cycle function and protein synthesis with and without this therapy. In addition to height and body weight, blood levels of proteins, lipids, growth hormone (GH), insulin-like growth factor-I (IGF-I) and IGF-binding protein -3 (IGFBP-3) were monitored. RESULTS: The frequency of hyperammonemic attacks and urinary orotate excretion decreased significantly following the resumption of arginine treatment. Despite showing no marked change in body weight, height increased gradually. Extremely low plasma arginine increased to normal levels, while plasma glutamine and alanine levels decreased considerably. Except for a slight increase in high-density lipoprotein cholesterol level, blood levels of markers for nutrition did not change. In contrast, low serum IGF-I and IGFBP-3 levels increased to age-matched control levels, and normal urinary GH secretion became greater than the level observed in the controls. CONCLUSION: Arginine treatment is able to reduces attacks of hyperammonemia in boys with late-onset OTCD and to increase their growth. PMID- 16703327 TI - Diabetic nephropathy is associated with increased albumin and fibrinogen production in patients with type 2 diabetes. AB - AIMS/HYPOTHESIS: Hyperfibrinogenaemia and albuminuria are cardiovascular risk factors, often coexisting in diabetic and non-diabetic people. Albuminuria in turn is associated with a compensatory albumin overproduction in non-diabetic patients. It is not known whether the presence of albuminuria in patients with type 2 diabetes mellitus is associated with greater albumin and fibrinogen production rates than in normoalbuminuric patients. SUBJECTS, MATERIALS, AND METHODS: Using leucine isotope methods, we measured fractional and absolute synthesis rates (FSR, ASR) of albumin and fibrinogen in post-absorptive type 2 diabetic patients with either normal (n=11) or increased (n=10) urinary albumin excretion. RESULTS: In albuminuric patients, albumin FSR (16.2+/-1.5%/day) and ASR (20.5+/-1.9 g/day) were greater (p<0.02 and p<0.05, respectively) than in normoalbuminuric patients (FSR=11.5+/-1.1%/day; ASR=15.7+/-1.2 g/day). Fibrinogen FSR was similar between patients with normal and increased albumin excretion, but concentration, the circulating pool and ASR of fibrinogen were 40 to 50% greater (p<0.035) in patients with albuminuria. Albuminuria was positively correlated with albumin ASR, with fibrinogen concentration, the fibrinogen pool and ASR, whereas albumin synthesis was inversely correlated with calculated oncotic pressure. CONCLUSIONS/INTERPRETATION: Synthesis of albumin and fibrinogen is upregulated in type 2 diabetic patients with increased urinary albumin excretion. Albuminuria is associated with enhanced fibrinogen and albumin synthesis. PMID- 16703328 TI - Reduced beta cell function in offspring of mothers with young-onset type 2 diabetes. AB - AIMS/HYPOTHESIS: Animal models indicate that even exposure to mild maternal hyperglycaemia in utero is detrimental to the beta cell function of the offspring, but evidence of this in humans is limited. In Europids who are diagnosed with type 2 diabetes before the age of 50 years, the risk of diabetes in the offspring of the diabetic mothers is greatly increased compared with the risk in those born to diabetic fathers. We hypothesised that offspring born to mothers with young-onset type 2 diabetes would have been exposed to mild hyperglycaemia in utero, so we studied the impact of this on their beta cell function. SUBJECTS AND METHODS: We measured beta cell function using early insulin response (EIR) after oral glucose; insulin resistance using HOMA; and HbA(1c) in 568 non-diabetic adult offspring born to parents with type 2 diabetes (mean age 55.8 years), split according to which parent was affected (in 327 it was the mother) and parental age of diagnosis: <50 years (n=117) or > or =50 years. To reduce the impact of genetic susceptibility, the offspring of affected fathers were used as control subjects. RESULTS: Offspring of mothers with young onset type 2 diabetes had lower EIR (log EIR 4.32, 95% CI [4.14-4.51] vs 4.63 [4.43-4.83] p=0.02) and higher HbA(1c) (4.89% [4.79-4.99] vs 4.68% [4.57-4.79] p=0.02) than the offspring of fathers with young-onset type 2 diabetes. Insulin sensitivity was similar in the two groups. There were no differences in EIR or HbA(1c) between the offspring born to mothers and fathers who were diagnosed after the age of 50 years. CONCLUSIONS/INTERPRETATION: We conclude that the offspring of mothers with young-onset type 2 diabetes have a reduction in beta cell function. This is consistent with exposure to mild maternal hyperglycaemia programming beta cell function. PMID- 16703329 TI - Voxel-based morphometry demonstrates reduced grey matter density on brain MRI in patients with diabetic retinopathy. AB - AIMS/HYPOTHESIS: In addition to nephropathy, retinopathy and peripheral neuropathy, a microvascular complication of type 1 diabetes that may be tentatively referred to as 'diabetic encephalopathy' has gained increasing attention. There is growing evidence that lowered cognitive performance in patients with type 1 diabetes is related to chronic hyperglycaemia rather than recurrent episodes of severe hypoglycaemia, as previously speculated. The aim of our study was to use magnetic resonance imaging (MRI) to establish whether long term hyperglycaemia, resulting in advanced retinopathy, contributes to structural changes in the brain (reduced grey matter). SUBJECTS, MATERIALS AND METHODS: We applied voxel-based morphometry on magnetic resonance images to compare grey matter density (GMD) between three groups of participants. GMD is used as a marker of cortical atrophy. We compared 13 type 1 diabetic patients with a microvascular complication (i.e. proliferative retinopathy) with 18 type 1 diabetic patients who did not have retinopathy in order to assess the effects of microvascular changes on GMD. Both patient groups were compared with 21 healthy control subjects to assess the effect of diabetes in itself. RESULTS: Patients with diabetic retinopathy showed reduced GMD in the right inferior frontal gyrus and right occipital lobe compared both with patients without retinopathy and with healthy controls (p<0.05). CONCLUSIONS/INTERPRETATION: Our data show that patients with type 1 diabetes, who, as a consequence of chronic hyperglycaemia, had developed advanced retinopathy, also showed increased focal cortical atrophy on brain MRI. PMID- 16703334 TI - Newspaper coverage of complementary and alternative therapies for cancer--UK 2002 2004. AB - BACKGROUND: Complementary and alternative therapies (CATs) are increasingly being used by cancer patients. These patients often rely on information retrieved from the lay press, which can affect their choices towards unconventional treatments for their disease. In this study, we aimed at assessing UK newspapers' coverage of CATs for cancer. METHODS: The "Lexis Nexis" database was searched for 3-month periods in 2002, 2003 and 2004. The search terms were combined: "complementary OR alternative AND medicine OR therapy AND cancer". CATs were grouped and articles' contents were assessed according to predefined criteria. RESULTS: A total of 310 articles were found: 117 came from national newspapers; 193 came from local newspaper issues. The UK press showed an increasing interest towards CATs for cancer (in 2002, 81 articles; in 2003, 82 articles and in 2004, 147 articles). The most frequently mentioned alternative therapies were diets and supplements (17.7%). Articles mainly focused on CATs as possible cancer treatments (44.8%), and 53.4% of all CATs mentioned were not backed up by trial data. The tone of the articles was generally positive towards CATs. Promotional articles increased over the years, especially for cancer centres and clinics. CONCLUSION: UK national newspapers frequently publish articles on CATs for cancer. Much of this information seems to be uncritical with a potential for misleading patients. PMID- 16703333 TI - Expectations to and evaluation of a palliative home-care team as seen by patients and carers. AB - OBJECTIVES: Although the number of palliative home-care teams is increasing, knowledge of what patients and principal informal carers expect from a home-care team is sparse. We aimed to elucidate this as well as evaluate a home-care team. PATIENTS AND METHODS: Individual semi-structured interviews with nine patients and six carers before receiving home care and 2-4 weeks after. In total, 26 interviews were conducted. Interviews were analysed with Template Analysis. Peer debriefing was performed. MAIN RESULTS: Patients and carers expected the team members to have specialised knowledge in palliative care and to improve their sense of security being at home. They also expected respite for carers and activities for patients. They evaluated the team positively but missed respite for carers and 24-h on-call service. CONCLUSIONS: Patients and carers found the home-care team essential to their sense of security being at home. Primary health care professionals must receive any necessary training outside patients' homes. Offering respite for carers and 24-h on-call service would be an improvement. PMID- 16703330 TI - Paternal insulin resistance and its association with umbilical cord insulin concentrations. AB - AIMS/HYPOTHESIS: Fetal growth is influenced by genetic factors as well as the intra-uterine environment. We hypothesised that some genetic factors may alter fetal insulin secretion and insulin action. SUBJECTS, MATERIALS AND METHODS: To assess this, we analysed plasma insulin concentration in umbilical cord blood from 644 normal, term, UK Caucasian deliveries from the Exeter Family Study of Childhood Health. We tested for associations between cord insulin and each of parental anthropometry, fasting glucose, insulin and lipids. RESULTS: As expected, cord insulin concentrations correlated with all measures of birth size (weight, length, head and arm circumferences, sum of skinfold thicknesses, ponderal index: r=0.16-0.4, p<0.01 for all) and maternal BMI (r=0.11, p=0.005), maternal glucose (r=0.25, p<0.001) and maternal insulin resistance (r=0.23, p<0.001). Paternal fasting insulin and insulin resistance were correlated with cord insulin (r=0.15, p=0.006; r=0.13, p=0.001, respectively), and this was independent of paternal BMI. Multiple linear regression analysis revealed paternal insulin resistance to be a predictor of cord insulin concentrations, independently of maternal factors. CONCLUSION: Our results show an independent relationship between paternal insulin resistance and cord insulin concentrations. This is consistent with heritability of insulin resistance from father to offspring and a compensatory increase in fetal insulin secretion, the latter occurring pre-natally before the homeostatic feedback loop between glucose and insulin is established. PMID- 16703335 TI - Inhibition of enamel lesion formation by fluoridated milk assessed by laser fluorescence--an in vitro study. AB - The aim of this study was to investigate the effect of fluoridated milk on enamel lesion formation as assessed by laser fluorescence (LF). The material consisted of 18 extracted premolar teeth that were cut in mesial-distal direction and pairwise assigned to either test or control samples in an experimental caries model. The teeth were exposed to a low-pH 5% cellulose gel for 4 h, 5 days per week immediately followed by a 4-h period in either fluoridated (5 ppm, test) or nonfluoridated milk (control). In the meantime, the specimens were stored in pooled human-stimulated whole saliva in room temperature. All teeth were examined by visual inspection with a magnifying glass and by LF readings (DIAGNOdent) at baseline and after 2 and 4 weeks. The baseline LF readings ranged from 3 to 7 with a mean value of 5.6+/-0.9. The mean values increased with time in both groups but the increase was more marked in the control teeth, 8.7+/-2.3 vs 12.8+/ 3.3 after 4 weeks, this difference being statistically significant (p<0.01). The visual examination could not distinguish between the test or control samples after 2 and 4 weeks, respectively. The findings indicated that fluoride added to milk may to some extent counteract enamel lesion formation as assessed by LF in an experimental caries model. PMID- 16703337 TI - Exercise preconditioning upregulates cerebral integrins and enhances cerebrovascular integrity in ischemic rats. AB - We hypothesized that exercise preconditioning strengthens brain microvascular integrity against ischemia/reperfusion injury through the tumor necrosis factor (TNF)-integrin signaling pathway. Adult male Sprague Dawley rats (n = 24) were studied in: (1) exercise (the animals run on a treadmill 30 min each day) for 3 weeks, (2) non-exercise. Six animals from each group (n = 12) were subjected to stroke, the remaining animals served as controls (n = 6 x 2). Brain infarction and edema were determined by Nissl staining. Cerebral integrin expression was detected by immunochemistry and stereological methods. In addition, we used flow cytometry to address the causal role of TNF-alpha in inducing the expression of integrins in the human umbilical vein endothelial cells under TNF-alpha or vascular endothelial growth factor (VEGF) pretreatment. Exercise reduces brain infarction and brain edema in stroke. Expressions of integrin subunit alpha(1), alpha(6), beta(1), and beta(4) were increased after exercise. Exercise preconditioning reversed stroke-reduced integrin expression. An in vitro study revealed a causal link between the gradual upregulation of TNF-alpha (rather than VEGF) and cellular expression of integrins. These results demonstrated an increase in cerebral expression of integrins and a decrease in brain injury from stroke after exercise preconditioning. The study suggests that upregulation of integrins during exercise enhances neurovascular integrity after stroke. The changes in integrins might be altered by TNF-alpha. PMID- 16703336 TI - Enamel matrix derivative alone or in combination with a bioactive glass in wide intrabony defects. AB - This controlled clinical study investigated the clinical and radiographic outcome of wide intrabony periodontal defects treated by enamel matrix derivatives alone or in combination with a bioactive glass over a period of 8 months. Twenty-three chronic periodontitis patients, who received initial therapy and had radiographical interproximal defects with an associated probing depth of 6 mm or more and an intrabony component of at least 4 mm, were included. Each of the patients, contributing at least one intrabony defect, was treated with either enamel matrix derivative alone (group 1, n=10) or the combination (group 2, n=13). In both groups, all clinical and radiographical parameters were improved. Groups 1 and 2 presented a mean pocket reduction of 5.03+/-0.89 and 5.73+/-0.80 mm, recession of 0.97+/-0.24 and 0.56+/-0.18 mm, relative attachment gain of 4.06+/-1.06 and 5.17+/-0.85 mm, and radiographic bone gain of 2.15+/-0.42 and 2.76+/-0.69 mm, respectively. An intergroup comparison revealed significant differences for all of the parameters, yielding a more favorable outcome towards the combined approach. Within the limits of the study, both treatments resulted in marked clinical and radiographical improvements, but combined treatment seemed to enhance the results in the treatment of wide intrabony defects. PMID- 16703338 TI - Koch's postulates and infectious proteins. AB - Koch's postulates were formulated in the late nineteenth century as guidelines for establishing that microbes cause specific diseases. Because the rules were developed for living agents--particularly bacteria--their applicability to inanimate pathogens such as viruses and infectious proteins has been problematic. The unorthodox mechanism by which prion diseases are transmitted, involving specific physicochemical characteristics of the protein as well as susceptibility traits of the host, has made these disorders refractory to analysis within the context of the original Koch's postulates. In addition, evidence is accumulating that other proteopathies, such as AA amyloidosis, apolipoprotein AII amyloidosis, and cerebral Abeta amyloidosis, can be induced in vulnerable recipients by cognate proteinaceous agents. In light of the salient differences in the mode of disease-transmission by microbes and proteins, we propose modifications of Koch's postulates that will specifically accommodate presumed infectious proteins. PMID- 16703340 TI - [New techniques for the diagnosis of osteoporosis]. AB - In this review article current developments and applications in quantitative osteoporosis imaging are presented. Developments in the field of DXA include geometrical parameters of the proximal femur such as the "hip axis length" and new ROIs to determine BMD. Advances in QCT are new volumetric techniques to quantify BMD at the lumbar spine and the proximal femur. In addition techniques to determine BMD in standard contrast-enhanced abdominal computed tomography studies are described. Currently with the new bone quality concept in full bloom techniques to quantify trabecular bone architecture as new surrogates of bone strength are of increasing significance. Spatial high-resolution techniques such as magnetic resonance imaging and new computed tomography techniques have shown their potential in assessing trabecular bone structure. In addition ultrasound is considered a low-cost technique to explore bone quality. PMID- 16703344 TI - Cerebral metabolite differences in adolescents with low birth weight: assessment with in vivo proton MR spectroscopy. AB - BACKGROUND: Children with very low birth weight (VLBW) have a significantly increased risk of later neurodevelopmental problems, while infants born small for gestational age (SGA) at term are also at some risk of developing neurological impairment. OBJECTIVE: To investigate possible brain metabolite differences in adolescents with VLBW, SGA at term and controls by proton in vivo magnetic resonance spectroscopy (MRS) at 1.5 T. MATERIALS AND METHODS: MR spectra were acquired from volumes localized in the left frontal lobe, containing mainly white matter (54 subjects). Peak areas of N-acetyl aspartate (NAA), choline (Cho) and creatine (Cr) were determined, and the peak area ratio of NAA to Cr, total Cho to Cr, or NAA to Cho calculated. Probabilistic neural network (PNN) analysis was performed utilizing the chemical shift region containing resonances from NAA, Cho and Cr as inputs. RESULTS: No significant difference in the peak area ratios could be found using the Kruskal-Wallis test. By application of PNN, a correct classification of 52 of the 54 adolescents with a sensitivity and specificity exceeding 93% for all groups was achieved. CONCLUSION: Small, yet systematic, differences in brain metabolite distribution among the groups were confirmed by PNN analysis. PMID- 16703352 TI - Biodegradation of hexahydro-1,3,5-trinitro-1,3,5-triazine by novel fungi isolated from unexploded ordnance contaminated marine sediment. AB - Undersea deposition of unexploded ordnance (UXO) constitutes a potential source of contamination of marine environments by hexahydro-1,3,5-trinitro-1,3,5 triazine (RDX) and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX). Using sediment from a coastal UXO field, Oahu Island, Hawaii, we isolated four novel aerobic RDX-degrading fungi HAW-OCF1, HAW-OCF2, HAW-OCF3 and HAW-OCF5, tentatively identified as members of Rhodotorula, Bullera, Acremonium and Penicillium, respectively. The four isolates mineralized 15-34% of RDX in 58 days as determined by liberated 14CO2. Subsequently we selected Acremonium to determine biotransformation pathway(s) of RDX in more details. When RDX (100 microM) was incubated with resting cells of Acremonium we detected methylenedinitramine (MEDINA), N2O and HCHO. Also we detected hexahydro-1-nitroso 3,5-dinitro-1,3,5-triazine (MNX) together with trace amounts of hexahydro-1,3 dinitroso-5-nitro-1,3,5-triazine (DNX) and hexahydro-1,3,5-trinitroso-1,3,5 triazine (TNX). Under the same conditions MNX produced N2O and HCHO together with trace amounts of DNX and TNX, but we were unable to detect MEDINA. TNX did not degrade with Acremonium. These experimental findings suggested that RDX degraded via at least two major initial routes; one route involved direct ring cleavage to MEDINA and another involved reduction to MNX prior to ring cleavage. Nitrite was only detected in trace amounts suggesting that degradation via initial denitration did take place but not significantly. Aerobic incubation of Acremonium in sediment contaminated with RDX led to enhanced removal of the nitramine. PMID- 16703357 TI - Heme oxygenase up-regulation in ultraviolet-B irradiated soybean plants involves reactive oxygen species. AB - Ultraviolet-B (UV-B) radiation has a negative impact on plant cells, and leads to the generation of reactive oxygen species (ROS). Heme oxygenase (HO, EC 1.14.99.3) plays a protective role against oxidative stress in mammals, but little is known about this issue in plants. Here, we report for the first time the response of HO in leaves of soybean (Glycine max L.) plants subjected to UV-B radiation. Under 7.5 and 15 kJ m(-2 )UV-B doses, HO, catalase (CAT, EC 1.11.1.6) and ascorbate peroxidase (APX, EC 1.11.1.11) activities were increased and the production of thiobarbituric acid reactive substances (TBARS) regain control values after 4 h of plant recuperation. Treatment with 30 kJ m(-2) UV-B provoked a decrease in these antioxidant enzyme activities. Immunoblot analysis showed a 4.3 and 3.7-fold increase in HO-1 protein expression after irradiation with 7.5 and 15 kJ m(-2), respectively. HO-1 transcript levels were enhanced (up to 77%) at these doses, as assessed by semi-quantitative RT-PCR. These data demonstrated that increased HO activity was associated with augmented protein expression and transcript levels. Plants pre-treated with the antioxidant ascorbic acid did not show the UV-B-induced up-regulation of HO-1 mRNA, but hydrogen peroxide treatment could mimic this reaction. Our results indicate that HO is up-regulated in a dose depending manner as a mechanism of cell protection against oxidative damage and that such response occurred as a consequence of HO-1 mRNA enhancement involving ROS. PMID- 16703358 TI - Cryptochrome photoreceptors cry1 and cry2 antagonistically regulate primary root elongation in Arabidopsis thaliana. AB - Cryptochromes are blue-light receptors controlling multiple aspects of plant growth and development. They are flavoproteins with significant homology to photolyases, but instead of repairing DNA they function by transducing blue light energy into a signal that can be recognized by the cellular signaling machinery. Here we report the effect of cry1 and cry2 blue light receptors on primary root growth in Arabidopsis thaliana seedlings, through analysis of both cryptochrome mutant and cryptochrome-overexpressing lines. Cry1 mutant seedlings show reduced root elongation in blue light while overexpressing seedlings show significantly increased elongation as compared to wild type controls. By contrast, the cry2 mutation has the opposite effect on root elongation growth as does cry1, demonstrating that cry1 and cry2 act antagonistically in this response pathway. The site of cryptochrome signal perception is within the shoot, and the inhibitor of auxin transport, 1-N-naphthylphthalamic acid, abolishes the differential effect of cryptochromes on root growth, suggesting the blue-light signal is transmitted from the shoot to the root by a mechanism that involves auxin. Primary root elongation in blue light may thereby involve interaction between cryptochrome and auxin signaling pathways. PMID- 16703359 TI - Idiopathic intracranial hypertension: the validity of cross-sectional neuroimaging signs. AB - The aim of this study was to evaluate the accuracy of previously reported neuroimaging signs in establishing or excluding the diagnosis of idiopathic intracranial hypertension (IIH). In a retrospective study, 30 patients with confirmed IIH and 56 controls were evaluated with brain magnetic resonance imaging. All examinations were evaluated in a blinded fashion by three neuroradiologists for the presence or absence of the 'traditional' signs of IIH: empty sella turcica, deformation of the pituitary, slit-like ventricles, 'tight' subarachnoid spaces, flattening of the posterior globe, protrusion of the optic nerve, enhancement of the optic nerve head, distension of the optic nerve sheath and vertical tortuosity of the optic nerve. Optic nerve protrusion and enhancement, slit-like ventricles and tight cerebrospinal fluid spaces were not significantly associated with IIH (P>0.05). Posterior globe flattening, optic nerve sheath distension, optic nerve tortuosity, pituitary deformity and empty sella turcica were significantly associated with IIH (P<0.05). However, most of these are not helpful in a clinical setting, with the exception of posterior globe flattening. This is the only sign that, if present, strongly suggests the diagnosis of IIH (specificity 100%, 95% CI 93.6% to 100%; sensitivity 43.5%, 95% CI 27.3% to 60.8%; positive likelihood ratio 49.7). The majority of the reported signs for IIH on cross-sectional imaging are not helpful in establishing or excluding the diagnosis of IIH, and are of no value in the clinical setting. Flattening of the posterior aspect of the globe is the only sign that, if present, is suggestive of the diagnosis of IIH. PMID- 16703361 TI - Cognitive changes after carotid artery stenting. AB - INTRODUCTION: We aimed to test changes in cognitive performance after carotid artery stenting (CAS). METHOD: Ten patients were neuropsychologically tested at least 24 h before and 48 h after CAS. To diminish thromboembolic events, we used a proximal protection device. The following neuropsychological tests were selected: The Mini Mental State Examination (MMSE), symbol digit test and subtests of the Consortium to Establish a Registry for Alzheimer's Disease (CERAD) battery (verbal fluency, constructional practice, word list memory and delayed recall). Affective state was determined by the Beck Depression Score (BDS). RESULTS: No patient suffered from depression (BDS <1) or dementia (MMSE 29.9+/-1.5). Nine of the ten patients (P=0.12) showed increased speed in the Number Connection Test (NCT) (corresponding to trail making test). Most patients showed better or similar results concerning delayed recall (P=0.31). No change was observed in the symbol digit test, word list memory, verbal fluency or constructional practice. Better results concerning NCT and delayed recall after carotid stenting might be due to improved brain perfusion. CONCLUSION: After CAS, cognitive and memory performance seem to improve. Further studies with different time intervals and more refined testing, as well as perfusion-weighted imaging, are needed. PMID- 16703360 TI - The intersubject and intrasubject reproducibility of FMRI activation during three encoding tasks: implications for clinical applications. AB - The goal of the present study was to evaluate the inter- and intrasubject reproducibility of FMRI activation for three memory encoding tasks previously used in the context of presurgical functional mapping. The primary region of interest (ROI) was the medial temporal lobe (MTL). Comparative ROIs included the inferior frontal and fusiform gyri which are less affected by susceptibility induced signal losses than the MTL regions. Eighteen subjects were scanned using three memory encoding paradigms: word-pair, pattern, and scene encoding. Nine subjects underwent repeat scanning. Intersubject reproducibility of FMRI activation was evaluated by examining the percent of subjects who showed activation within a given ROI and the range to which individual laterality indices (LIs) varied from the mean. Intrasubject test-retest reproducibility was evaluated by examining the LI test-retest correlation, the average difference between LIs from two separate imaging sessions, and concordance ratios of activation volumes (R(volume) and R(overlap)). For scene encoding the reproducibility of activation volume and LIs within the MTL were as good as or better than the reproducibility within the fusiform and inferior frontal ROIs. For pattern encoding and word-pair encoding, the reproducibility of activation volume and LIs within the MTL tended to be worse compared to the fusiform and inferior frontal ROIs. The differences in FMRI reproducibility appeared more dependent on the task than the susceptibility effects. The results of this study suggest that FMRI-based assessment of the neural substrates of memory using a scene encoding task may be a useful clinical tool. PMID- 16703364 TI - Submillimetre spatial resolution is feasible in positron emission tomography. PMID- 16703365 TI - Detection of widespread neurolymphomatosis with 18F-FDG PET. PMID- 16703363 TI - Analysis of papaya BAC end sequences reveals first insights into the organization of a fruit tree genome. AB - Papaya (Carica papaya L.) is a major tree fruit crop of tropical and subtropical regions with an estimated genome size of 372 Mbp. We present the analysis of 4.7% of the papaya genome based on BAC end sequences (BESs) representing 17 million high-quality bases. Microsatellites discovered in 5,452 BESs and flanking primer sequences are available to papaya breeding programs at http://www.genomics.hawaii.edu/papaya/BES . Sixteen percent of BESs contain plant repeat elements, the vast majority (83.3%) of which are class I retrotransposons. Several novel papaya-specific repeats were identified. Approximately 19.1% of the BESs have homology to Arabidopsis cDNA. Increasing numbers of completely sequenced plant genomes and BES projects enable novel approaches to comparative plant genomics. Paired BESs of Carica, Arabidopsis, Populus, Brassica and Lycopersicon were mapped onto the completed genomes of Arabidopsis and Populus. In general the level of microsynteny was highest between closely related organisms. However, papaya revealed a higher degree of apparent synteny with the more distantly related poplar than with the more closely related Arabidopsis. This, as well as significant colinearity observed between peach and poplar genome sequences, support recent observations of frequent genome rearrangements in the Arabidopsis lineage and suggest that the poplar genome sequence may be more useful for elucidating the papaya and other rosid genomes. These insights will play a critical role in selecting species and sequencing strategies that will optimally represent crop genomes in sequence databases. PMID- 16703366 TI - Biological imaging for selecting and monitoring cancer therapy; a pathway to individualised therapy. PMID- 16703367 TI - Compensatory renal growth protects mice against Shiga toxin 2-induced toxicity. AB - Uninephrectomy (Unx) is followed by the compensatory renal growth (CRG) of the remaining kidney. Previous evidence has shown that during CRG, renal tissue is resistant to a variety of pathologies. We tested the hypothesis that the functional changes that take place during CRG could attenuate Shiga toxin (Stx) toxicity in a mouse model of Stx2-induced hemolytic uremic syndrome (HUS). The participation of nitric oxide (NO) was analyzed. After CRG induction with Unx, mice were exposed to a lethal dose of Stx2, and the degree of renal damage and mortality was measured. Stx2 effects on the growth, renal blood flow (RBF) and NO synthase (NOS) intrarenal expression in the remaining kidney were then studied. The induction of CRG strongly prevented Stx2-mediated mortality and renal damage. Administration of the NOS inhibitor NG-nitro-L-arginine methyl ester (L-NAME) during CRG partially impaired the protection. Both Stx2 and L-NAME interfered with the hypertrophic and hyperplastic responses to Unx, as well as with the increase in RBF. In intact mice, Stx2 decreased renal perfusion, inhibited endothelial NOS basal expression and enhanced inducible NOS expression; all of these effects were attenuated by prior Unx. It is concluded that during CRG mice are highly protected against Stx2 toxicity and lethality. The protective capacity of CRG could be related to the enhancement of renal perfusion and preservation of eNOS renal expression, counterbalancing two major pathogenic mechanisms of Stx2. PMID- 16703370 TI - International Vesicoureteral Reflux Study: unsolved questions remaining. PMID- 16703371 TI - Severe lactic acidosis and nephrolithiasis in an infant--etiology?: type 1 glycogen storage disease (GSD). PMID- 16703373 TI - Analysis of a uteroglobin gene polymorphism in childhood Henoch-Schonlein purpura. AB - Uteroglobin (UG) is a pleiotropic protein with anti-inflammatory properties. Mice rendered genetically incapable of expressing UG develop a form of renal disease that closely resembles human IgA nephropathy (IgAN). Furthermore, a single nucleotide polymorphism in the UG gene (A38G) has been associated with rapid progression of human IgAN. We examined whether the A38G polymorphism is associated with childhood Henoch-Schonlein purpura (HSP), a form of vasculitis associated with IgAN-like renal disease. We examined the prevalence of the A38G polymorphism in 34 children with HSP and in 38 ethnically matched controls. Only one patient had clinically evident renal involvement. As compared with controls, the prevalence of the 38G allele was slightly increased in children with HSP, but this increase was not statistically significant. Our results do not support a role for UG in susceptibility to childhood HSP in the population studied. Larger studies involving more patients with renal disease will be necessary to define whether UG is associated with increased risk for HSP nephritis. PMID- 16703374 TI - Influence of oral intake of Saccharomyces boulardii on Escherichia coli in enteric flora. AB - Enteric flora constitutes 95% of the cells in the human body. It has been shown that the bacterial content of this flora is affected by diet and changes in nutrition. Considering that urinary tract infections (UTI) are mostly due to ascending infections from the gut flora, the importance of the elements of this flora and their characteristics becomes more evident. The aim of this study was to evaluate the influence of oral Saccharomyces boulardii (S. boulardii) intake on the number of Escherichia coli (E. coli) colonies in the colon. This study was carried out with 14 boys and 10 girls (total of 24 children) aged between 36 and 192 months (mean: 104.3+/-45.1 months). A commercial capsule or powder containing 5 billion colony-forming units (cfu) of S. boulardii was administered once a day for 5 days. The number of E. coli and yeast colonies was measured in the stool samples of the study group before and after the use of this drug. Before treatment, the mean number of E. coli colonies in g/ml stool was 384,625+/ 445,744. This number decreased significantly to 6,283+/-20,283 after treatment (p=0.00). S. boulardii was not detected in stool before treatment and the number of colonies increased to 11,047+/-26,754 in g/ml stool. S. boulardii may be effective in reducing the number of E. coli colonies in stool. The influence of this finding on clinical practice such as prevention of UTI needs to be clarified by further studies. PMID- 16703375 TI - Treatment of primary and secondary hypertension in children. AB - The incidence of pediatric hypertension (HTN) is increasing, mainly due to an increase in primary (essential) HTN, or PH. There are only a limited number of studies assessing the characteristics and treatment efficacy of PH versus secondary HTN (SH). We conducted a retrospective analysis of 158 pediatric patients (mean age: 10.8 years; sex ratio: 51.1% female, 48.9% male) with HTN of whom 34.4% had PH and 65.6% had SH. The vast majority were either African American or Caucasian. Among all patients, therapy induced a significant decrease in systolic blood pressure (SBP) and diastolic BP (DBP) (both p<0.0001). SBP (p<0.0001) and DBP (p=0.002) declined significantly in PH patients. PH and SH patients with a body mass index (BMI) >95th percentile (%) had a significantly higher post-therapy SBP (both p<0.05) than those with a BMI <95th%. SBP declined similarly in PH patients treated with calcium-channel blockers (CCB) and angiotensin-converting enzyme inhibitors (ACEI). DBP declined only in PH patients treated with ACEI. SBP and DBP (both p<0.0001) declined significantly in SH patients. Post-therapy BP was similar in SH patients treated with either CCB or ACEI. Post-therapy SBP and DBP were significantly lower in SH patients than in PH patients; moreover, therapy induced a greater decline in SBP and DBP in the SH patients. Compared to PH patients, SH patients were twofold more likely to achieve a SBP less than the 95th% after therapy. We conclude that (1) significant lowering of BP with either CCB or ACEI is achievable in most children with HTN, and (2) SH patients respond better to therapy than those with PH. PMID- 16703376 TI - Health-related quality of life in pediatric patients with ESRD. AB - As part of creating a pediatric ESRD-specific Health-Related Quality of Life (HRQOL) assessment instrument, we established pilot data with the PedsQL 4.0 Generic Core Scales in 96 pediatric patients with ESRD receiving hemodialysis (HD), peritoneal dialysis (PD) or with a renal transplant (TX). Patient age ranges were: 2-4 (8; PD n=3, TX n=5), 5-7 (12; HD n=9, TX n=3), 8-12 (25; HD n=5, PD n=6, TX n=14), 13-18 (51; HD n=18, PD n=10, TX n=23). PedsQL ESRD data were compared to healthy children (n=131 child report; n=145 parent report) and across HD/PD/TX. For all domains, ESRD patient HRQOL scores were significantly lower than healthy controls. Transplant patients reported better physical and psychosocial health than dialysis patients. No difference was noted between HD and PD patients for any PedsQL domain. Our data demonstrate that the PedsQL 4.0 Generic Core Scales is a useful measurement instrument to screen for HRQOL impairment in pediatric patients with ESRD. We suggest that the ESRD-specific HRQOL instrument in development is needed to help define condition-specific HRQOL differences between modalities. PMID- 16703377 TI - Four-year follow-up of oral health surveillance in renal transplant children. AB - To outline the specific oral surveillance needs of renal transplant children, we report the 4-year follow-up data of 106 children examined routinely on a 6-month basis and upon request from the medical team or the parents in the interval. Data were recorded for hard and soft dental tissues, including enamel defects carious lesions, gingival status, orthodontic treatment needs, and wisdom teeth eruption. Hard-tissue lesions were noted in 34.9% of the children, caries lesions on the permanent teeth were seen in 15.0% of cases, 18.86% of the patients had orthodontic treatment. Spontaneous examination, upon request for medical reasons, was given for 26 children. Twenty-one demands originated from the parents. Over a 4-year period the attendance of a dental specialist resulted in a significant improvement in the oral health of the renal transplant children. PMID- 16703378 TI - Neonatal nephrotic presentation of a child with heterozygous NPHS1 mutation. AB - Congenital nephrotic syndrome of the Finnish type, due to homozygous mutation of NPHS1, is the most common form of congenital nephrotic syndrome. Angiotensin converting enzyme (ACE) and prostaglandin synthesis inhibition along with supportive albumin infusion therapy, with or without unilateral nephrectomy, has allowed management of the disease without dialysis until transplantation in some cases of congenital nephrotic syndrome. Reported here is a case of heterozygous NPHS1 mutation, with normal NPHS2 gene structure, presenting during prenatal screening and developing nephrotic syndrome within days of birth. The patient has responded well to very low doses of ACE inhibitors and indomethacin alone. This case illustrates the importance of an initial trial of conservative medical therapy in milder presentations of the congenital nephrotic syndrome, especially given the current limitations of diagnostic testing and our inadequate knowledge of the complete spectrum of disorders of podocyte proteins. PMID- 16703379 TI - Crescentic glomerulonephritis in a child with infective endocarditis. AB - Renal manifestations associated with infective endocarditis (IE) may present with different clinical patterns, and the most common renal histopathological finding is diffuse proliferative and exudative type of glomerulonephritis, leading to hematuria and/or proteinuria. Renal failure due to crescentic glomerulonephritis (CGN) in children with IE is a very rare condition. We report here a 6-year-old boy, who had a history of cardiac surgery for pulmonary atresia and ventricular septal defect, presenting with the clinical findings of IE and hematuria associated with renal failure due to CGN. He was treated with a combination of intravenous (IV) methylprednisolone pulses and appropriate antibiotics, but also received one dose of IV cyclophosphamide. Complete serological, biochemical, and clinical improvement was achieved after 2 months of follow-up. Antibiotic therapy is the essential part of the treatment of IE-associated glomerulonephritis; however, this case also highlights the importance of aggressive immunosuppressive therapy to suppress the immunological process related with infection in this life threatening condition leading to renal failure. PMID- 16703380 TI - Persistent post-transplant polyuria managed by bilateral native-kidney laparoscopic nephrectomy. AB - Polyuria is not considered an absolute indication for pre-transplant nephrectomy; however, it may complicate post-transplantation fluid management. Bilateral native-kidney laparoscopic nephrectomy was performed at our centre in two patients (four kidneys) 1 month after they had received a living related-donor renal transplant. The indication for nephrectomy was severe post-transplant polyuria secondary to the patient's underlying disease: juvenile nephronophthisis. Both patients had a persistent post-transplant daily urine output of 7-8 l/day and continued to have a variable serum creatinine level, dependent on intravenous hydration, more then 3 weeks after transplantation. Bilateral laparoscopic native-kidney nephrectomy in children has previously been reported. However, to the best of our knowledge, laparoscopic nephrectomy has not been described after kidney transplantation and certainly not in the immediate post-transplantation period. The procedure was well tolerated and did not affect renal graft function. In fact, following the procedure, serum creatinine levels stabilized, while daily fluid requirements decreased to 2.5-3.5 l/day in both patients. We concluded that bilateral native-kidney nephrectomy can be safely performed in paediatric renal transplant recipients in the immediate post transplantation period. This new approach may allow preemptive transplantation and avoid the need for a transition period on dialysis in patients for whom pre transplant nephrectomy is not absolutely indicated. PMID- 16703381 TI - [Antibiotic therapy: progress and development of resistance]. AB - This contribution illustrates reasons, spread, and mechanisms of development of resistance especially in gram-positive microorganisms such as methicillin resistant staphylococcus aureus (MRSA) or vancomycin resistant enterokoccus (VRE). Possibilities of overcoming these are described as well as strategies for antibacterial therapy with recently developed antibiotics against multiple resistant microorganisms. These are linezolid as an oxazolidinone, daptomycin as a lipopeptid and tigecyclin as a new glycylcycline, which are already on the market or will be launched soon in Germany. Differences in magnitude and frequency of still existing resistances between hospital and practice are discussed with respect to their importance for the internal physician. PMID- 16703385 TI - Electrophoretic analysis of proteins and enantiomers using capillaries modified by a successive multiple ionic-polymer layer (SMIL) coating technique. AB - The applicability of three-layer coatings consisting of three different polymers (A(+)-B(-)-C(+) coating) prepared by a successive multiple ionic-polymer layer (SMIL) coating technique to the immobilization of polypeptides and/or proteins onto the inner surface of the capillaries was investigated to provide a high performance separation medium for proteins and enantiomers in capillary electrophoresis (CE). To obtain a stable protein-coated capillary, high molecular mass poly(ethyleneimine) (PEI) was employed as the first layer in the A(+)-B(-) C(+) coating, and then a cationic protein was immobilized as the third layer. Comparisons of analytical performances between the A(+)-B(-)-C(+) coating and the conventional SMIL-coated (A(+)-B(-)-A(+) coating) capillary were conducted. The CE separation of cationic proteins was successfully achieved with the prepared capillaries. In addition, the polypeptide- and protein-coated capillaries were applied to the chiral separation of a binaphthyl compound. It should be noted that the chiral separation efficiency was strongly dependent on the second anionic polymer layer of the coating. Effects of the interaction between oppositely charged ionic polymer layers on the separation efficiency are discussed. PMID- 16703383 TI - DFNB68, a novel autosomal recessive non-syndromic hearing impairment locus at chromosomal region 19p13.2. AB - From a large collection of families with autosomal recessive non-syndromic hearing impairment (NSHI) from Pakistan, linkage has been established for two unrelated consanguineous families to 19p13.2. This new locus was assigned the name DFNB68. A 10 cM genome scan and additional fine mapping were carried out using microsatellite marker loci. Linkage was established for both families to DFNB68 with maximum multipoint LOD scores of 4.8 and 4.6. The overlap of the homozygous regions between the two families was bounded by D19S586 and D19S584, which limits the locus interval to 1.9 cM and contains 1.4 Mb. The genes CTL2, KEAP1 and CDKN2D were screened but were negative for functional sequence variants. PMID- 16703386 TI - Symptomatic palatal myoclonus with ear click after tick-borne meningoencephalitis. AB - We present a case of a 39-year-old patient, who was diagnosed and treated for a tick-borne meningoencephalitis. Three months after the treatment he started to complain of annoying, cracks-resembling, rhythmical sounds, coming from the inside of his head to both his ears. Physical examination revealed rhythmical oscillations of the soft palate with a frequency of 100-120 per minute and a clock ticking noise synchronic with the palate tremor. Electromyography revealed continuous motor unit activity at rest in the tensor veli palatini muscle. Palatal myoclonus (PM) as a result of tick-borne meningoencephalitis was diagnosed. Treatment with several medications was started with no effect, then botulinum toxin was administered under EMG guidance to both sides of the patient's soft palate with great improvement. A 5-year follow-up and continuation of botulinum toxin injections with only minor and reversible side effects proved the treatment efficacy and safety. In the article we present a case of symptomatic palatal myoclonus with ear click and shortly discuss its aetiology, types and treatment options. We also stress the efficacy and safety of PM treatment with repetitive injections of botulinum toxin. PMID- 16703387 TI - Low prevalence of transfusion transmitted virus (TTV)-like DNA sequences in cystadenolymphoma and pleomorphic adenoma of the salivary glands. AB - Titers of transfusion transmitted virus (TTV)-like DNA in saliva samples have been reported 100-1,000 times higher than those of the corresponding sera, suggesting viral transmission by saliva droplets. The present study was conducted to determine whether TTV-like DNA sequence elements play a role in the pathogenesis of cystadenolymphoma or pleomorphic adenoma and if the parotid or the submandibular gland is a major source of TTV persistence. Sixty-two archival salivary gland samples (16 cystadenolymphomas, 13 pleomorphic adenomas, and 33 controls) and 23 corresponding saliva samples were examined using a polymerase chain reaction (PCR) for TTV DNA. All PCR products that displayed DNA bands were sequenced. Leder's stain and immunohistochemistry (anti-CD8, anti-CD20, anti CD45R0, anti-CD68, and anti-Ki67/MiB1) were applied to detect possible changes associated with findings of TTV-like DNA sequences. Tissue displayed TTV-like DNA sequences in 8.1% (5/62; saliva: 47.8%, 11/23). Tissue that contained TTV-like DNA sequences was histologically indistinguishable from samples lacking such DNA. TTV appears to be only a bystander in cystadenolymphoma, pleomorphic adenoma, and other salivary gland affections. Neither of the glands seems to be a major source of TTV persistence. PMID- 16703388 TI - Distribution of pendrin in the organ of Corti of mice observed by electron immunomicroscopy. AB - The distribution of pendrin, which is encoded by the Pendred syndrome gene, has been investigated immunohistochemically in the inner ear. In the cochlea, pendrin has been found in the spiral prominence, external sulcus cells, Hensen's cells and Claudius cells, but its expression in the organ of Corti remains unclear. We examined whether pendrin localizes in the organ of Corti by postembedding immunogold analysis. In the organ of Corti, gold particles were clearly observed in outer and inner hair cells, including the stereocilia. The density of the particles was especially high in the cuticular plates of the hair cells. Gold particles were also detected in the external sulcus, in part of the spiral ligament adjacent to the external sulcus, in supporting cells, and in the spiral ganglion of the cochlea. Our study revealed that pendrin occurs in the organ of Corti. The role of pendrin in the organ of Corti and its association with the Cl- or pH regulation of neurotransmission require further study. PMID- 16703389 TI - Aroclor 1254 impairs the hearing ability of Xenopus laevis. AB - In this study we assessed the effects of chronic, dietary exposure of Aroclor 1254 (A1254) on the hearing of Xenopus frogs. We used the auditory brainstem response (ABR) to assay changes in hearing physiology; ABR thresholds, as well as latency-intensity and amplitude-intensity profiles of the initial positive (P1) and negative (N1) peaks were measured. Two groups of animals that received 50 ppm and 100 ppm of A1254 in their diet from 5 days post-fertilization through metamorphosis were compared to a control group that received untreated chow. The results showed significant threshold elevations in the 3-4 kHz range and significantly delayed peak latencies and reduced amplitudes at these frequencies in A1254 treated animals as compared to control animals. These findings indicate that A1254 selectively damages the high-frequency sensorineural hearing system associated with the basilar papilla of frogs. This preferential damage may be related to inherent differences in the vulnerability of the basilar versus amphibian papilla in the frog. The overall results of this study are also consistent with the reported A1254-induced auditory deficits in mammals indicating that the basilar papilla of the Xenopus frog may serve as an effective model for studying the effects of A1254 on the auditory system. PMID- 16703391 TI - Percutaneous vertebroplasty or kyphoplasty: which one do I choose? PMID- 16703390 TI - On the barn owl's visual pre-attack behavior: I. Structure of head movements and motion patterns. AB - Barn owls exhibit a rich repertoire of head movements before taking off for prey capture. These movements occur mainly at light levels that allow for the visual detection of prey. To investigate these movements and their functional relevance, we filmed the pre-attack behavior of barn owls. Off-line image analysis enabled reconstruction of all six degrees of freedom of head movements. Three categories of head movements were observed: fixations, head translations and head rotations. The observed rotations contained a translational component. Head rotations did not follow Listing's law, but could be well described by a second-order surface, which indicated that they are in close agreement with Donder's law. Head translations did not contain any significant rotational components. Translations were further segmented into straight-line and curved paths. Translations along an axis perpendicular to the line of sight were similar to peering movements observed in other animals. We suggest that these basic motion elements (fixations, head rotations, translations along a straight line, and translation along a curved trajectory) may be combined to form longer and more complex behavior. We speculate that these head movements mainly underlie estimation of distance during prey capture. PMID- 16703392 TI - How does practice reduce dual-task interference: integration, automatization, or just stage-shortening? AB - The present study assessed three hypotheses of how practice reduces dual-task interference: Practice teaches participants to efficiently integrate performance of a task pair; practice promotes automatization of individual tasks, allowing the central bottleneck to be bypassed; practice leaves the bottleneck intact but shorter in duration. These hypotheses were tested in two transfer-of-training experiments. Participants received one of three training types (Task 1 only, or Task 2 only, or dual-task), followed by dual-task test sessions. Practice effects in Experiment 1 (Task 1: auditory-vocal; Task 2: visual-manual) were fully explained by the intact bottleneck hypothesis, without task integration or automatization. This hypothesis also accounted well for the majority of participants when the task order was reversed (Experiment 2). In this case, however, there were multiple indicators that several participants had succeeded in eliminating the bottleneck by automatizing one or both tasks. Neither experiment provided any evidence that practice promotes efficient task integration. PMID- 16703394 TI - [Functional anatomy of the abdominal wall]. AB - The anterolateral abdominal wall covers a region defined cranially by the xiphoid process and ribs, laterally by the medial axillary line, and caudally by the anterior ilium and pubic bone. Knowledge of the various parts of the abdominal wall is essential to the surgeon for effective laparotomy and primary and secondary hernia care. The abdominal musculature, aponeuroses, vascularity, and innervation are examined in detail along with the according vascular and neural structures of the dermis. PMID- 16703395 TI - [Incision and closure of the abdominal wall]. AB - Access to the abdominal cavity must be performed in such a way that surgical treatment procedures can be performed safely. For skin incision, scalpel and electrocautery are equivalent. Subcutaneous tissue and fascias must be divided by electrocautery to minimize blood loss. The best way to close the abdominal cavity is by an all layer, slowly absorbable, running suture with a suture: wound length ratio of at least 4:1. Closing the peritoneal layer is not necessary. Subcutaneous sutures and drains do not reduce the risk of wound complications. Staples should be used for closing the skin. PMID- 16703396 TI - [Treatment of abdominal wall defects, including abdominal relaxation]. AB - Abdominal wall defects that are amenable to direct fascial approximation are the indication for retromuscular mesh augmentation. Larger defects can be bridged with meshes used as abdominal wall substitution or with reconstructive flaps. Other options are indirect techniques to achieve primary preparation of the abdominal wall, either by mobilization of the abdominal wall muscles with the component separation method or by preoperative expansion of the abdominal cavity using a progressive pneumoperitoneum. Surgical repair of iatrogenic abdominal wall relaxations should combine plastic reconstruction with preperitoneal mesh implantation. Scientific evidence supporting any treatment option is weak, because few prospective, randomized trial data are available owing to the inhomogeneity of the patient population. Treatment of abdominal wall defects must therefore be proposed on an individual basis utilizing one or a combination of the techniques described. PMID- 16703397 TI - [Abdominal hernias resulting from abdominal flap harvest]. AB - The anatomic characteristics of the anterior abdominal wall allow the harvesting of various types of flaps, some of them with considerable volume. These flaps are used mainly for reconstruction of the female breast, thoracic wall, and perineal or ilioinguinal region. Even though general donor site morbidity is low, hernias and "bulging" can occur due to the harvest of muscle and fascia, which leads to a weakening of the abdominal wall. Hernias and bulging appear mostly after harvest of classic flap types, which include removal of the rectus abdominis muscle. Further refinements of these flaps, i.e. microvascular flaps, in particular perforator flaps, lead to a marked reduction in donor site morbidity. In the following overview, the problem of abdominal wall weakening as a result of flap harvest is discussed and possible therapeutic options are elucidated. PMID- 16703399 TI - The group II metabotropic glutamate receptor agonist, LY379268, inhibits both cocaine- and food-seeking behavior in rats. AB - RATIONALE: Group II metabotropic glutamate receptor (mGluR2/3) agonists are proposed to serve as potential treatment for addiction. OBJECTIVES: The present study examined the hypothesis that mGluR2/3 agonists exert inhibitory effects on cocaine-induced reinstatement of cocaine-seeking. METHODS: Rats were trained to self-administer either cocaine or control reinforcer (food), then responding on the reinforcer-paired lever was extinguished. Reinstatement of responding was induced by a noncontingent presentation of the self-administered reinforcer (10 mg/kg cocaine, i.p. or 765 mg of food). In one experiment, rats were systemically pretreated with vehicle (Veh) or the mGluR2/3 agonist LY379268 (0.3, 1, or 3 mg/kg, i.p.) 30 min before the reinstatement test session. In a second experiment, Veh or LY379268 (0.05, 0.5, or 5 nmol/side) was microinjected into the nucleus accumbens core (NAc core) 5 min before the reinstatement test session. The effects of LY379268 on cocaine- and food-induced reinstatement on reward seeking were assessed. RESULTS: Both systemic and intra-NAc core pretreatment with LY379268 inhibited both cocaine- and food-seeking behavior. However, the effect of LY379268 appeared somewhat more effective for cocaine seeking than food-seeking. CONCLUSIONS: These results support a potential therapeutic role for mGluR2/3 agonists on relapse of cocaine-seeking. However, doses that inhibited cocaine-seeking were only threefold lower than those inhibiting food-seeking, indicating possible unacceptable nonspecific effects. In addition, the NAc core is one site of action where the mGluR2/3 agonists elicit effects on reward-seeking behavior. PMID- 16703398 TI - Involvement of c-myc-regulated genes in hepatocellular carcinoma related to genotype-C hepatitis B virus. AB - PURPOSE: The purpose of this study was to elucidate the molecular basis of hepatocellular carcinoma (HCC) caused by genotype-C hepatitis B virus (HBV). METHODS: We compared molecular profiles of 15 HCCs and five non-tumorous livers, all of which were associated with genotype-C HBV infection, using DNA microarray technology. RESULTS: Our supervised learning identified 237 genes whose expression differed between HCCs and non-tumorous livers. This result was validated by a false discovery rate of 0%. Levels of expression of 35 and 202 genes were higher and lower, respectively, in HCCs than in non-tumorous livers. Among the 237 genes, we highlighted the top 35 upregulated and top 35 down regulated genes in tumor. Interestingly, when overlapping genes were excluded, 12 (e.g., NM23-H2, MCM7, PARP1, YWHAH, HSPB1, and MSH2) of the top 34 upregulated genes and five (e.g., MT1A and MT3) of the top 33 downregulated genes were c-myc regulated genes. The microarray data for five randomly selected genes (MCM7, UBE2L3, PPIA, CXCL12, and ASS) were confirmed by quantitative real-time reverse transcription-polymerase chain reaction. CONCLUSIONS: Our results indicate that many c-myc-regulated genes are involved in genotype-C-HBV-related HCC, suggesting that c-myc is related to the hepatocarcinogenic activity of genotype-C HBV. PMID- 16703400 TI - Metabolic correlates of toluene abuse: decline and recovery of function in adolescent animals. AB - RATIONALE: Children and adolescents will readily abuse household products that contain solvents such as toluene. It is likely that reinforcing exposures to toluene alter brain glucose metabolism. OBJECTIVE: Using an animal model of drug reinforcement, we sought to identify a metabolic signature of toluene abuse in the adolescent rodent brain. Small animal PET (microPET), in combination with the glucose analog radiotracer, (18)FDG, were used to evaluate the metabolic consequences of inhaled toluene. METHODS: The exposure protocol paralleled our previously established method for assessing the conditioned reinforcing effects of toluene (5,000 ppm) using the conditioned place preference (CPP) paradigm. Animals were scanned at baseline and 2 h after the last exposure. Follow-up (18)FDG scans occurred 1 day, 3 weeks, and 2 months later. RESULTS: After six pairings, 38% of the animals preferred the toluene paired chamber and 25% were averse. The immediate metabolic effect in toluene-exposed animals was a 20% decline in whole brain (18)FDG uptake. Twenty-four hours following the last exposure, the whole brain decline was 40%, and 2 months later, the decline was 30% of pretoluene levels. A region-by-region analysis demonstrated significant additional decreases in the pons, cerebellum, striatum, midbrain, temporal cortex, and hippocampus. Two months after toluene cessation, regions of complete metabolic recovery were the thalamus and cerebellum; however, the temporal cortex did not recover. CONCLUSIONS: Brain uptake of (18)FDG appears to be a useful tool for examining the metabolic impact of toluene abuse, which include a profound decline followed by region-specific recovery after cessation. PMID- 16703401 TI - Dopamine D4 receptor polymorphism modulates cue-elicited heroin craving in Chinese. AB - RATIONALE: Subjective craving, which contributes to the continuation of drug use in active abuser and the occurrence of relapse in detoxified abusers, is considered to be a central phenomenon in addiction. Dopamine pathway has been implicated in the mechanism underlying the cue-elicited craving for a variety of addictive substances. OBJECTIVES: The objective of this study was to test the hypothesis that heroin addicts carrying D4 dopamine receptor gene (DRD4) variable number tandem repeat (VNTR) long type allele would have higher craving after exposure to a heroin-related cue. MATERIALS AND METHODS: Craving was induced by a series of exposure to neutral and heroin-related cue and were assessed in a cohort of Chinese heroin abusers (n=420) recruited from the Voluntary Drug Dependence Treatment Center at Shanghai. RESULTS: Significantly stronger cue elicited heroin craving was found in individuals carrying DRD4 VNTR long type allele than the non-carriers (F=31.040, p<0.001). As for baseline craving and mean change in craving responding to neutral stimuli, no significance was found (1.06+/-0.34 vs 1.07+/-0.36, F=0.067, p=0.797 and 0.42+/-0.34 vs 0.45+/-0.37, F=0.277, p=0.599, respectively). CONCLUSIONS: The results of our study suggest that DRD4 VNTR polymorphism contributes to cue-elicited craving in heroin dependence, indicating DRD4 VNTR represents one of potential genetic risk factors for cue-induced craving. PMID- 16703402 TI - The comparative study of Sprague-Dawley and Lewis rats in adjuvant-induced arthritis. AB - The outbred Sprague-Dawley (SD) rats, similar to the inbred Lewis (LEW) rats, have been recently demonstrated to be highly susceptible to adjuvant-induced arthritis (AIA). We herein compared AIA in SD and LEW rats in terms of clinical, histological, radiological, and immuno-inflammatory features. The results showed that, following inoculation with a ground Mycobacterium tuberculosis (MT) suspension, SD and LEW rats manifested closely similar disease progression, with 100% incidence and similar severity. The development of arthritis was accompanied by significantly higher erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) levels than in control rats. Radiographic examination of the hind paws showed that both SD and LEW AIA rats manifested conspicuous soft tissue swelling, bone matrix resorption, periosteal new bone formation and bone erosion, while histopathological analysis of the synovial joints revealed marked cellular infiltration, angiogenesis, synovial hyperplasia, pannus formation, narrowing of joint space, and cartilage and bone destruction. Moreover, in relation to disease progression, serum tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL 1beta), and IL-6 levels were markedly overexpressed in both SD and LEW AIA versus control rats, and SD and LEW AIA rats exhibited divergent profiles for the expression of TNF-alpha and IL-1beta. Taken together, these results demonstrated that the SD rat AIA model shares several arthritic features with the comparable model in LEW rats. Hence, given the more favorable characteristics of SD rats than LEW rats (i.e., lower cost, wider availability, and heterogenic background), this SD rat AIA model is more cost effective and advantageous for screening and testing novel anti-arthritic agents. PMID- 16703403 TI - Growth hormone: historical notes. AB - A brief review of important contributions to our present knowledge of growth hormone is given. In 1887 it had been noted that a pituitary tumor was present in most patients with acromegaly. Even at the beginning of the 20. Century relationship between growth disorders and the pituitary was contested. From 1908 pituitary surgery became established treatment in GH hypersecretion. In 1922 it was demonstrated that injection of pituitary extract to animals caused excessive growth and soon after the opposite: removal of the pituitary caused growth retardation. A huge number of studies on the effects of GH were subsequently reported as were trials with GH treatment. They were impeded by failure to recognize the impact of species specificity of GH. After this issue was clarified in 1957, treatment with human growth hormone proved effective. In 1985 it was realized that Creutzfeldt-Jakob's disease might be transmitted through human growth hormone. At this time recombinant GH had become available. In 1971 the structure of human GH was established. In the same period both GH releasing and inhibiting hormones were identified and an analogue of somatostatin had evolved into the first effective pharmacological treatment for acromegaly. PMID- 16703404 TI - Pituitary tumors arising from glycoprotein hormone alpha-subunit-deficient mice contain transcription factors and receptors present in thyrotropes. AB - Glycoprotein-hormone alpha-subunit deficient (alphaSUnull) mice are hypothyroid and hypogonadal due to the absence of functional TSH, LH and FSH, despite normal production of the corresponding beta subunits. Pituitary tumors spontaneously developing in alphaSUnull mice were propagated in hypothyroid mice. The purpose of the current studies was to compare the gene expression profile of these alphaSUnull tumors with previously characterized TtT-97 thyrotropic tumors. A group of animals bearing each tumor type was treated with thyroid hormone (T4) prior to tumor removal. Both tumor types equally expressed TSHbeta mRNA, which significantly decreased when exposed to T4, whereas alpha-subunit mRNA was absent in alphaSUnull tumors. Northern blot analysis was performed using cDNA probes for the following transcription factors: Pit1, GATA2, pLIM, Msx1, Ptx1 and Ptx2. Both tumors were found to contain identical transcripts with similar responses to T4, with the exception of Pit1. In contrast to the signal pattern seen in TtT-97, only two bands were seen in alphaSUnull tumors, which were similar in size to those in alphaTSH cells, a thyrotropic cell line that lacks TSHbeta-subunit expression and Pit1 protein. However, western blot analysis revealed a protein band in the alphaSUnull tumors consistent with Pit1, while this signal was absent in alphaTSH cells. Northern blot analysis was also performed with specific cDNA probes for the following receptors: TRbeta1, TRbeta2, TRalpha1, non-T3 binding alpha2, RXRgamma and Sst5. Similarly-sized transcripts were found in both types of tumor, although the signal for Sst5 was seen in T4-treated alphaSUnull tumors only with a more sensitive RT-PCR analysis. The overall similarity between the two tumor types renders the alphaSUnull tumor as a suitable thyrotropic tumor model. PMID- 16703405 TI - Weight and dimensions of the pituitary in northwestern Indians. AB - Weight and dimensions of the pituitary (hypophysis cerebri) obtained from medicolegal autopsies of northwestern Indian subjects, which included 87 children and adolescents and 798 adults were recorded. Volume of the pituitary was determined in 100 specimens. In addition, anteroposterior and vertical measurement of pituitary were taken in mid-sagittal sections of the head in magnetic resonance images (MRI) of 130 living adults. In the males, the weight of pituitary increased steadily from 102.52 +/- 38.66 mg in the age group of 0-5 years to 427.83 +/- 117.15 mg in the age group of 36-45 years, it decreased thereafter. In the females, the weight increased from 166.10 +/- 38.70 mg in the first age group to 445.90 +/- 168.60 mg in the age group of 16-17 years and became erratic thereafter. The mean weight of the gland in female subjects was always more than in the males of the corresponding ages till 35 years (p < 0.001, p < 0.01, p < 0.05). The maximum weight of the pituitary was observed during adolescence in the females. When weights of the gland of all the age groups were pooled together in adults the average weight was 401.26 +/- 105.89 mg in the males and 417.32 +/- 104.07 mg in the females. The weight and dimensions of the gland in northwestern Indian subjects were smaller than those in the western Caucasians and Japanese. In mid-sagittal MRI pictures of the head, the anteroposterior and vertical measurement of pituitary were about one mm greater than in the autopsy specimens. In the males, weight of the gland was significantly related to body weight in children, adolescents and adults; it was related to supine body length only in the adults. In the females, weight of the gland was significantly correlated with age in all the age group except in the age group of 36-55 years. PMID- 16703407 TI - Lymphocytic hypophysitis: disease spectrum and approach to diagnosis and therapy. AB - Lymphocytic hypophysitis (LYH) is a neuroendocrine disorder characterized by autoimmune inflammation of the pituitary gland with various degrees of pituitary dysfunction. The histopathology consists of an initial monoclonal lymphocytic infiltrate, which can heal with minimal sequela or progress to fibrosis and result in permanent hypopituitarism. Coexistence of other autoimmune conditions is reported in 25-50% of cases and pituitary autoantibodies have been detected in up to 70% of biopsy-proven cases. The clinical presentation varies depending on the pituitary segment that is more severely affected. In lymphocytic adenohypophysitis (LAH) an early destruction of the ACTH-producing cells is characteristic. Other anterior pituitary hormones can also be affected but posterior pituitary involvement is absent or minimum. Lymphocytic Infundibuloneurohypophysitis (LINH) typically presents as acute onset diabetes insipidus (DI) with intracranial mass-effect symptoms. A combination of extensive anterior pituitary involvement and DI characterizes lymphocytic Infudibulopanhypophysitis (LIPH). The diagnosis can be challenging in many cases, because distinction from pituitary adenomas and other sellar masses is not obvious. Significant efforts have been made to identify specific serum markers, but it would seem unlikely that this approach will ever have the specificity to replace histopathological examination of a surgical specimen. Diagnostic criteria have been proposed to help in the decision-making process and to avoid, whenever possible, unnecessary invasive procedures. The therapeutic approach is controversial and, although transsphenoidal surgery is often performed, a conservative medical management is justified in many cases, given the self limited nature of the inflammatory process. This paper reviews the etiology, epidemiology, clinical and radiological findings, diagnosis and management of LYH. PMID- 16703408 TI - Waxing and waning of a pituitary mass in a young woman with combined pituitary hormone deficiency (CPHD) due to a PROP-1 mutation. AB - We describe a 23-yr old woman with congenital combined pituitary hormone deficiency (CPHD) diagnosed at 10 years of age and a large sellar mass discovered at the age of 19 years, when her first pituitary MRI was performed. The mass (height: 13 mm) extended to the suprasellar region, close to the optic chiasm, showed signal hyperintensity in T1- and hypointensity in T2-weighted images, with no enhancement after gadolinium injection. Although these MRI features were suggestive of Rathke's cleft cyst, cystic craniopharyngioma or previous hemorrhage, no visual symptoms, diabetes insipidus and/or hyperprolactinemia were present. In addition, similar MRI findings had been previously described in a few cases of CPHD due to PROP-1 mutations, which prompted us to carry out a molecular study before any therapeutic decision was made. A 301302delAG PROP-1 mutation was found in her DNA and the patient was closely followed through ophthalmologic evaluation and pituitary MRI scans. During a 3.6-year follow-up, we were able to document a marked initial growth followed by shrinkage and recurrent growth of the PROP-1 sellar mass. The patient remains free of compressive neuro ophthalmological signs, suggesting that surgical intervention is unnecessary in these cases. However, they must be followed closely with sellar MRIs and campimetry until the mass completely regresses. PMID- 16703409 TI - Pituicytoma presenting with spontaneous hemorrhage. AB - Pituicytomas are rare tumors. Previously reported pituicytomas all presented with signs and symptoms relating to mass effect or endocrinological dysfunction. We report a 47 year old man who presented with sudden, severe headache and was found to have a hemorrhagic suprasellar mass with hemorrhage into the third ventricle. A mass arising from the pituitary stalk was found at surgery, and thorough pathological analysis revealed a pituicytoma. Pituicytoma should be considered in the differential diagnosis of a hemorrhagic suprasellar mass. PMID- 16703406 TI - Antiproliferative effects of somatostatin analogs in pituitary adenomas. AB - The antisecretory effects of somatostatin (SRIF) and its analogs are widely recognised and provide the basis for treatment of hormonal hypersecretion in pituitary adenomas, especially in the settings of acromegaly. Evidence for an antiproliferative effect of these compounds has also been provided. This review focuses on the mechanisms transducing the antiproliferative effects of SRIF and its analogs on pituitary adenomas, and on the clinical consequences on tumor volume of pharmacological treatment of pituitary adenomas with these drugs. PMID- 16703410 TI - Adipsic diabetes insipidus following pituitary surgery for a macroprolactinoma. AB - Adipsic diabetes insipidus (ADI) is a rare condition in which thirst, an essential clinical feature for the prevention of hypernatraemic dehydration, is absent. We report the first case of adipsic diabetes insipidus to occur following surgery for a pituitary macroprolactinoma, with loss of both osmoregulated and baroregulated vasopressin release. Following extensive surgery for a vision threatening macroprolactinoma a 14-year-old boy developed profound hypernatraemia with absent thirst sensation. Detailed investigation, with hypertonic saline infusion and trimetaphan infusion, revealed absence of both osmoregulatory and baroregulatory release of vasopressin. We discuss the investigation and management of such patients and the physiology of hypothalamic-neurohypophyseal dysfunction in such patients. PMID- 16703411 TI - Intrasellar plasmacytoma presenting as a non-functional invasive pituitary macro adenoma: case report & literature review. AB - We report an uncommon case of an intrasellar plasmacytoma presenting as a non functional invasive pituitary macro-adenoma as the first manifestation of multiple myeloma. A 57 year old woman was referred to our department with a presumed diagnosis of a non-functioning pituitary macro-adenoma. She reported a 3 month history of headaches and a 2 week history of sudden onset of right facial numbness. Preoperative endocrine evaluation was remarkable only for a modestly elevated serum prolactin. A magnetic resonance imaging (MRI) scan revealed 3.6 x 5 x 4 cm enhancing homogeneous intrasellar mass with extension into the sphenoid and cavernous sinuses bilaterally; the optic chiasm was not displaced. She underwent transphenoidal surgery of the sellar lesion. The surgical specimen was heavily infiltrated with abnormal plasma cells, which stained almost exclusively for Kappa light chain immunoglobulins. An extensive investigation was undertaken to look for occult myelomatous disease. A diagnosis of multiple myeloma was made 1 month later based on a combination of clinical, pathological and radiologic features. She underwent radiation therapy directed towards the residual sellar tumor, followed by chemotherapy and autologous stem cell transplantation. Review of the world literature revealed only 22 previous reports of patients in whom a solitary plasmacytoma or multiple myeloma first presented as a sellar mass; in all cases mimicking clinically and radiologically a non-functioning invasive pituitary adenoma however with additional cranial nerve involvement. Intrasellar plasma cell tumors are rare tumors which may mimic non-functioning invasive pituitary tumors. The diagnosis should be suspected in patients with well preserved anterior pituitary function and cranial nerve neuropathies in the presence of significant sellar destruction. PMID- 16703412 TI - Suprasellar tubercular abscess presenting as panhypopituitarism: a common lesion in an uncommon site with a brief review of literature. AB - Following widespread use of imaging, detection rate of abnormal sites of parenchymal neuro-tuberculosis is on a rise. A handful of cases of tuberculomas/abscesses in hypothalamo-pituitary region have been reported and most of them are diagnosed on surgical histopathology. We describe a patient of suprasellar tubercular abscess, who presented with visual disturbances, diabetes insipidus with panhypopituitarism and on histopathology had granulomas and positive acid fast bacilli. PMID- 16703414 TI - Reduced MTA1 expression by RNAi inhibits in vitro invasion and migration of esophageal squamous cell carcinoma cell line. AB - To distinguish aggressive esophageal squamous cell carcinoma from indolent disease is the important clinical challenge. Studies have indicated that metastasis-associated gene 1(Mta1) played a role in the process of metastasis of carcinoma. The overexpression of Mta1 gene has been found in a variety of tumors. To identify the detailed roles of MTA1 protein in the carcinogenesis of esophageal squamous cell carcinoma, this study analyzed the pathological specimens on tissue microarray derived from 72 patients using immunohistochemistry. MTA1 expression increased in the nuclear with the development of esophageal squamous cell carcinoma from normal epithelial cell, dysplasia, to invasive cancer. In biological studies with human esophageal squamous cell carcinoma cell line, MTA1 plays its roles to promote cancer cell invasion, adhesion and movement. RNA interference (RNAi) against MTA1 decreased the malignant phenotypes. Gene microarray analysis revealed some metastasis associated genes were altered by MTA1 RNAi. This study started an effective beginning to explore metastasis mechanisms and cancer gene therapy strategy targeting MTA1. PMID- 16703413 TI - Assessing tumor growth and distribution in a model of prostate cancer metastasis using bioluminescence imaging. AB - Bioluminescence imaging (BLI) has greatly facilitated the development of animal models of cancer, allowing sensitive detection of luciferase-expressing cancer cells in living mice. Previous efforts characterizing such models have involved small numbers of animals, limiting understanding of their performance features. We employed BLI to serially image the growth and distribution of a prostate cancer cell line, 22Rv1, after intracardiac injection into scid mice (n = 85). This approach models hematogenous dissemination of cancer cells and allows inquiry of the process of metastatic colonization at various organ sites, although accurately injecting cancer cells into the left ventricle remains challenging. Therefore, to predict injection success we measured the ratio of the thoracic bioluminescence signal to the whole body bioluminescence signal (T/WB ratio) immediately following intracardiac injection. A T/WB ratio less than 0.50 predicted the development of tumors outside of the thoracic cavity while a T/WB greater than 0.50 predicted the development of tumors entirely within the thoracic cavity, suggestive of a failed injection. Progressive tumor growth was quantified using BLI. Tumors colonized multiple organ sites including bone, liver, and adrenal glands resembling the spectrum of metastases in autopsy studies of patients with prostate cancer. Tumors growing in bone exhibited mixed osteolytic and osteoblastic features, eliciting a spiculated periosteal response. With the ability to more accurately predict injection success, we can now monitor efficacy of intracardiac injections facilitating the performance of this model. PMID- 16703416 TI - Impact of CCR5 Delta32/+ deletion on herpes zoster among HIV-1-infected homosexual men. AB - The association between the presence of CCR5 Delta32 heterozygosity and incidence of clinical herpes zoster was studied among 296 homosexual men from the Amsterdam cohort study (ACS) infected with human immunodeficiency virus type I (HIV-1) with an estimated date of seroconversion. Of them 63 were CCR5 Delta32 heterozygotes and 233 CCR5 wild-type. The incidence rate of a first episode of herpes zoster was 4.2% and 5.3% per person-year, respectively. A higher occurrence of herpes zoster was strongly related to a lower CD4 + cell count. After adjustment for age, presence of CCR2b 64I heterozygosity, HIV RNA load, time since seroconversion, and CD4 + cell count, the rate ratio for herpes zoster of CCR5 Delta32 was 0.9 (95%CI 0.5-1.6). In conclusion, in HIV-1-infected homosexual men, a CCR5 Delta32 heterozygous genotype has no protective effect on the incidence of herpes zoster. PMID- 16703417 TI - IL-20: biological functions and clinical implications. AB - IL-20 belongs to the IL-10 family and plays a role in skin inflammation and the development of hematopoietic cells. Little is known about its other biological functions and clinical implications, however. Updated information about IL-20, such as its identification, expression, receptors, signaling, biological activities, and potential clinical implications, is illustrated in this review based on our research and on data available in the literature. Our studies of IL 20 show that it is a pleiotropic cytokine with potent inflammatory, angiogenic, and chemoattractive characteristics. Inflammation and angiogenesis are essential for the pathogenesis of rheumatoid arthritis and atherosclerosis. Based on in vitro data and clinical samples, we demonstrated that IL-20 is involved in the diseases of rheumatoid arthritis and atherosclerosis. In addition, we found in our studies that IL-20 signaled through different molecules in several cells. The present review presents the clinical implications of IL-20 in rheumatoid arthritis and atherosclerosis. It may provide new therapeutic options in the future. PMID- 16703418 TI - Possible routes for lead accumulation in feral pigeons (Columba livia). AB - This study examined possible routes for lead (Pb) accumulation in resident pigeons collected from rural, urban, and four industrial sites in Korea. The accumulation pattern of Pb was comparable to the study sites. The highest Pb concentration was found in the bone, followed by kidney, liver, and lung of pigeons. Highest Pb residues in bones were found in urban (Seoul), and two industrial complex areas (Busan and Ulsan), which were about 15 times higher than rural area (Duckjuk island), and followed by Ansan and Yochon industrial areas. Regional Pb variations in liver, kidney, and lung tissues were also similar pattern with the bone Pb difference. These findings indicate that Pb accumulation in tissues of pigeons may be affected by the Pb exposure in their respective habitats. Crop contents and gizzard materials were investigated as representing the ingested items. No difference of Pb concentration was observed in major foods (maize and/or wheat) of crop contents in the study sites except Busan, whereas variations of Pb levels in gizzard materials were indicative of a similar pattern with tissue Pb differences. The Pb concentration in tissues of pigeons did not correspond well to the atmospheric Pb levels. With regard to possible Pb sources, ingested items especially materials present in the gizzard are important sources for Pb contamination to pigeons because Pb-containing sources may be expected to present in roadside particles, dusts, paint chips and building flakes. However, air Pb value being low may not affect significant as the regional variations in tissues of pigeons. PMID- 16703415 TI - Translation of 'rare' codons in a cell-free protein synthesis system from Escherichia coli. AB - We analyzed the effect of nine 'rare' codons (AGA, AGG, AUA, CCC, CGA, CGG, CUA, GGA, and UUA) on gene expression in an Escherichia coli coupled transcription/translation cell-free system, in comparison with a cell-based expression system. Each reporter gene contained five consecutive repeats of a rare codon, or in some experiments, three consecutive repeats. The cell-free expression of the genes bearing the codons CGA, CUA, GGA, and UUA was not affected, although these codons, except for GGA, were inefficiently translated in E. coli cells. Translation of the remaining five codons (AGA, AGG, AUA, CCC, and CGG) was severely reduced in both systems, and was remarkably facilitated in the cell-free system based on an S30 extract from the E. coli cells overproducing 'minor' tRNAs for these codons. PMID- 16703419 TI - Resonance assignments for the hypothetical protein TA0938 from Thermoplasma acidophilum. PMID- 16703420 TI - Sequence specific resonance assignment of a hypothetical protein PA0128 from Pseudomonas aeruginosa. PMID- 16703421 TI - Backbone resonance assignment of human adult hemoglobin in the deoxy form. PMID- 16703422 TI - Pulse transit time based on piezoelectric technique at the radial artery. AB - OBJECTIVES: Pulse transit time (PTT) has shown its potential in relevant cardiovascular and cardiorespiratory studies. However, the use of photoplethysmography (PPG) in PTT measurement can be limited in events of poor peripheral perfusion. Uninterrupted PTT monitoring may also not be achievable when less cooperative patients distribute the PPG probe due to its prominent light source. Hence, there is a need for an alternative method to measure PTT in such incidents. METHODS: In this study, the piezoelectric (PIEZO) technique to detect pulsations from a human wrist above the radial artery to estimate PTT is presented. 17 healthy adults (11 male; age range of 21-33 years) were recruited to compare PTT and heart rate (HR) differences between the PPG and PIEZO methods. These time-related derivations were made with respect to an electrocardiogram (ECG). RESULTS: The timing consistency of the PIEZO transducer shows significant correlations (p < 0.01) to those derived from the ECG and a pulse oximeter. Particularly, there is a high level of agreement of < 1 beat per minute (bpm) difference in HR estimates observed when compared to the two commercial devices in the respective Bland-Altman plots. Comparison of PTT obtained from the PIEZO transducer against the PPG signal shows constantly lower values due to the shorter path length it requires to propagate. A regression equation was formulated to relate the PTT values acquired from both these signals. CONCLUSIONS: Preliminary findings herein suggest that the PIEZO technique can be useful as an alternative for PTT monitoring. This shows promise to be more accommodating for less cooperative patients or those with insufficient peripheral perfusion. PMID- 16703426 TI - Preoperative evaluation of the extrahepatic bile duct structure for laparoscopic cholecystectomy. AB - BACKGROUND: The incidence of aberrant bile duct injury associated with laparoscopic cholecystectomy (LC) has not yet been adequately examined. This study aimed to clarify the types of normal cystic ducts and the incidence of aberrant extrahepatic bile ducts, and to search for a method of avoiding injuries during LC. METHODS: Aberrant hepatic ducts were retrospectively categorized into five types according to the pattern of the cystic ducts and the accessory hepatic ducts by preoperative endoscopic retrograde cholangiography or multidetector three-dimensional computed tomography using drip infusion cholangiography. The aberrant bile ducts were classified as type A (merging at the right side of the common bile duct), type B (merging at the anterior side), or type C (merging at the posterior left side). RESULTS: The intrahepatic bile ducts and cystic duct were clearly shown for 1,044 of the 1,278 patients who underwent LC. Secondary branches of aberrant cystic ducts were observed in 37 cases (3.5%), and accessory hepatic ducts were observed in 30 cases (2.9%). A comparison of the difficulties encountered with LC for each type based on the merging patterns of cystic ducts showed that type C needed a much longer operation time for LC than the other types. CONCLUSIONS: A preoperative evaluation of the bile duct tract and the accessory hepatic duct before LC is important. Patients with a cystic duct merging normally into the posterior left side of the common hepatic duct (type C) experienced difficulty when undergoing LC. The authors have safely performed LC with the use of an endoscopic nasobiliary drainage tube in type D cases (cystic duct merging with the right hepatic duct), in type IV cases (cystic duct merging with an accessory hepatic duct). PMID- 16703428 TI - Endoscopic band ligation therapy for upper gastrointestinal bleeding related to Mallory-Weiss syndrome. AB - BACKGROUND: No consensus exists as to the best endoscopic treatment for Mallory Weiss syndrome. Endoscopic band ligation is a readily available and easily learned technique. This prospective study evaluated the efficacy and safety of endoscopic band ligation therapy for Mallory-Weiss syndrome. METHODS: From August 1998 to June 2005, a clinical trial assessed 37 patients with a diagnosis of Mallory-Weiss syndrome who had active bleeding, exposed vessels, or both. Their lesions were treated using endoscopic band ligation. RESULTS: Endoscopic band ligation was successful in 36 of 37 cases, with a follow-up period ranging from 1 to 24 months. The remaining patient had severe liver failure and disseminated intravascular coagulation. The patient bled again at 12 h and subsequently died. Except for this case, no recurrent bleeding, perforation, or other complications occurred. CONCLUSIONS: The study results suggest that endoscopic band ligation is an effective, safe, and easily learned procedure for treating upper gastrointestinal bleeding related to Mallory-Weiss syndrome. PMID- 16703427 TI - First experience with robot-assisted thoracoscopic esophagolymphadenectomy for esophageal cancer. AB - BACKGROUND: Transthoracic esophagectomy with extended lymph node dissection is associated with higher morbidity rates than transhiatal esophagectomy. This morbidity rate could be reduced by the use of minimally invasive techniques. The feasibility of robot-assisted thoracoscopic esophagectomy (RTE) with mediastinal lymphadenectomy was assessed prospectively. METHODS: This study investigated 21 consecutive patients with esophageal cancer who underwent RTE using the Da Vinci robotic system. Continuity was restored with a gastric conduit and a cervical anastomosis. RESULTS: A total of 18 (86%) procedures were completed thoracoscopically. The operating time for the thoracoscopic phase was 180 min (range, 120-240 min), and the median blood loss was 400 ml (range, 150-700 ml). A median of 20 (range, 9-30) lymph nodes were retrieved. The median intensive care unit stay was 4 days (range, 1-129 days), and the hospital stay was 18 days (range, 11-182 days). Pulmonary complications occurred in 10 patients (48%), and one patient (5%) died of a tracheoneoesophageal fistula. CONCLUSIONS: In this initial experience, robot-assisted thoracoscopic esophagectomy was found to be feasible, providing an effective lymphadenectomy with low blood loss. Standardization of the technique and increased experience should reduce the complication rate, which is in the range of the rate for open transthoracic dissection. PMID- 16703429 TI - Complementary use of local excision and transanal endoscopic microsurgery for rectal cancer after neoadjuvant chemoradiation. AB - BACKGROUND: Neoadjuvant therapies have significantly improved local control and survival of patients with rectal cancer. Nevertheless, although a complete pathologic response can be achieved in 30% of cases, a transabdominal surgical resection is always required. This study aimed, for the first time, to test in the literature the feasibility of local excision combined with transanal endoscopic microsurgery (TEM) as a surgical option for patients treated with neoadjuvant chemoradiation. METHODS: Between July 1997 and December 2002, 30 patients with rectal cancer affected by an extraperitoneal tumor entered a protocol consisting of neoadjuvant chemoradiation followed by surgery. The surgical treatment, consisting of open surgery, local excision, or TEM, was planned according to the patient's clinical response after chemoradiation and distance from the anal verge. RESULTS: A significant clinical downstaging was observed in eight patients. Five of these patients underwent TEM, and three had local excision. Consequently, open surgery was performed for 22 patients. Histology showed six cases of complete pathologic response: three in the open surgery group and three in the transanal excision group. After a mean follow-up period of 47 months, the disease-free survival rate was 77% in the open surgery group and 100% in TEM or local excision group. CONCLUSIONS: The findings suggest the complementary feasibility of TEM and local excision after neoadjuvant chemoradiation. However, randomized trials are needed to confirm the oncologic safety of this approach. PMID- 16703431 TI - Does open surgery continue to have a role in elective splenectomy? AB - BACKGROUND: Since it was first reported in 1991, laparoscopic splenectomy has become the procedure of choice for elective splenectomy. However, doubts have been raised regarding the suitability of patients with splenomegaly (>1 kg) for laparoscopic resection because there have been reports of greater morbidity and higher conversion rates in this group of patients. Since 2000, patients referred to the authors' center for splenectomy with an estimated spleen weight exceeding 1 kg have undergone splenectomy by the open approach. METHODS: Between September 1995 and April 2005, 95 elective splenectomies were performed by a single surgeon. Operative data were collected prospectively. RESULTS: A comparison between the operations that took place before 2001 (n = 47) and those performed after 2000 (n = 48) for all sizes of spleen showed significant reductions in conversion rate, operative time, and hospital stay in the later group. As compared with laparoscopic splenectomy (n = 11), open splenectomy (n = 18) for cases of splenomegaly resulted in a significantly shorter operative time, less operative blood loss, and no significant difference in hospital stay. CONCLUSION: Although laparoscopic splenectomy is the treatment of choice for the majority of patients requiring elective splenectomy, the procedure for patients with significant splenomegaly requires caution and common sense. This study shows that an open splenectomy for these patients significantly reduces operative time and blood loss without increasing morbidity or hospital stay. PMID- 16703430 TI - Laparoscopic versus open ventral hernia mesh repair: a prospective study. AB - BACKGROUND: An incisional hernia develops in 3% to 13% of laparotomy incisions, with primary suture repair of ventral hernias yielding unsatisfactory results. The introduction of a prosthetic mesh to ensure abdominal wall strength without tension has decreased the recurrence rate, but open repair requires significant soft tissue dissection in tissues that are already of poor quality as well as flap creation, increasing complication rates and affecting the recurrence rate. A minimally invasive approach was applied to the repair pf ventral hernias, with the expectation of earlier recovery, fewer postoperative complications, and decreased recurrence rates. This prospective study was performed to objectively analyze and compare the outcomes after open and laparoscopic ventral hernia repair. METHODS: The outcomes for 50 unselected patients who underwent laparoscopic ventral hernia repair were compared with those for 50 consecutive unselected patients who underwent open repair. The open surgical operations were performed by the Rives and Stoppa technique using prosthetic mesh, whereas the laparoscopic repairs were performed using the intraperitoneal onlay mesh (IPOM) repair technique in all cases. RESULTS: The study group consisted of 100 patients (82 women and 18 men) with a mean age of 55.25 years (range, 30-83 years). The patients in the two groups were comparable at baseline in terms of sex, presenting complaints, and comorbid conditions. The patients in laparoscopic group had larger defects (93.96 vs 55.88 cm2; p = 0.0023). The mean follow-up time was 20.8 months (95% confidence interval [CI], 18.5640-23.0227 months). The mean surgery durations were 90.6 min for the laparoscopic repair and 93.3 min for the open repair (p = 0.769, nonsignificant difference). The mean postoperative stay was shorter for the laparoscopic group than for the open hernia group (2.7 vs 4.7 days; p = 0.044). The pain scores were similar in the two groups at 24 and 48 h, but significantly less at 72 h in the laparoscopic group (mean visual analog scale score, 2.9412 vs 4.1702; p = 0.001). There were fewer complications (24%) and recurrences (2%) among the patients who underwent laparoscopic repair than among those who had open repair (30% and 10%, respectively). CONCLUSIONS: The findings demonstrate that laparoscopic ventral hernia repair in our experience was safe and resulted in shorter operative time, fewer complications, shorter hospital stays, and less recurrence. Hence, it should be considered as the procedure of choice for ventral hernia repair. PMID- 16703432 TI - Laparoscopic myomectomy for large uterine fibroids. A comparative study. AB - BACKGROUND: The goal of this study was to examine the safety and feasibility of laparoscopic myomectomy (LM) for the management of symptomatic intramural uterine fibroids with weight greater than 80 g as compared to those less than 80 g. METHODS: In a prospective comparative study, 176 women with symptomatic uterine fibroids were scheduled for LM. They were divided into two groups, one with main uterine fibroid (intramural type) weight greater than 80 g and the other with fibroid weight less than 80 g. Outcome measures for the two groups were studied in terms of operation time, amount of blood loss, requirement of blood transfusion, and length of hospital stay. RESULT: Operation time and amount of blood loss were significantly greater in the group with fibroid > or = 80 g than in the group < 80 g (121.5 +/- 58.9 min versus 79.1 +/- 28.6 min, p < 0.001; and 346.3 +/- 299.6 ml versus 123.0 +/- 89.7 ml, p < 0.001, respectively). However, there was no difference in the length of hospital stay and overall incidence of operative complications between these two groups. None of the women had any major complications. Nevertheless, 11 minor complications were noted, including two pelvic abscesses requiring a second laparoscopic treatment. There was no incidence of switching to laparatomy during the operation. Extreme intraoperative hemorrhage of more than 1000 ml occurred in 8 patients; however, all progressed to full recovery after blood transfusion. Rate of blood transfusion was significantly lower in the group with fibroid < 80 g (3.2% versus 22.1%, p < 0.001). CONCLUSIONS: Despite the increased operation time and blood loss, LM can be safely performed in the treatment of large uterine fibroid. However, high risk of blood transfusion in these patients has to be kept in mind. PMID- 16703433 TI - Effect of prolonged warm ischemia and pneumoperitoneum on renal function in a rat syngeneic kidney transplantation model. AB - BACKGROUND: Laparoscopic donor nephrectomy is associated with several advantages for the donor. However, graft function may be impaired due to use of pneumoperitoneum and prolonged warm ischemia. This study investigated the impact of pneumoperitoneum and prolonged warm ischemia on long-term graft function in a syngeneic rat renal transplant model. METHODS: A total of 27 Brown Norway rats were randomized for transplantation of kidneys after three different procedures: no insufflation and no warm ischemia (group 1), no insufflation with 20 min of warm ischemia (group 2), and CO2 insufflation and 20 min of warm ischemia (group 3). Glomerular filtration rate (GRF), serum creatinine, urine volume, urine creatinine, and proteinuria were determined monthly for 1 year. One year after transplantation, the grafts were removed for histomorphologic analysis. RESULTS: No significant differences in GRF, serum creatinine, urine volume, and proteinuria were found among the three groups. Histologic analysis also showed no differences between the groups. CONCLUSION: Warm ischemia in combination with CO2 pneumoperitoneum, as used in laparoscopic donor nephrectomy, does not result in a negative effect on long-term graft function. PMID- 16703434 TI - Superiority of preemptive analgesia with intraperitoneal instillation of bupivacaine before rather than after the creation of pneumoperitoneum for laparoscopic cholecystectomy: a randomized, double-blind, placebo-controlled study. AB - BACKGROUND: This study aimed to evaluate the optimal timing of preemptive analgesia with bupivacaine peritoneal instillation in a prospective, randomized, double-blind, placebo-controlled trial. METHODS: In this study, 120 patients qualified for laparoscopic cholecystectomy were randomized to four groups. Group A received 2 mg/kg of bupivacaine in 200 ml of normal saline before creation of pneumoperitoneum. Group B received 2 mg/kg of bupivacaine in 200 ml of normal saline after creation of pneumoperitoneum. Group C received 200 ml of normal saline before creation of pneumoperitoneum. Group D received 200 ml of normal saline after creation of pneumoperitoneum. Local wound infiltration with bupivacaine was used before skin incisions. The primary end points of the study were postoperative pain intensity on a visual analog scale and incidence of shoulder tip pain. The secondary end points included the latency of nurse controlled analgesia activation, the analgesia request rate, and analgesic consumption. RESULTS: Significantly lower visual analog scores were observed in group A versus groups C and B versus group D during the initial 48 and 24 h, respectively. The patients in group A versus group B reported significantly lower pain at 4 h (p < 0.001) and 8 h (p = 0.003) postoperatively, but the difference was not significant after 12, 24, and 48 h. None of the group A patients reported shoulder tip pain, whereas it was reported by 3 patients in group B, 6 patients in group C, and 7 patients in group D (p < 0.01). The latency of nurse-controlled analgesia activation was 426.8 +/-57.2 min in group A, as compared with 307 +/- 39.8 min in group B, 109.3 +/- 51 min in group C, and 109 +/- 46.5 min in group D (p < 0.001). A significantly lower analgesia request rate was observed in group A versus C, as compared with group B versus D, throughout the entire study period (p < 0.05). CONCLUSIONS: Preemptive analgesia with bupivacaine peritoneal instillation is much more effective for pain relief if used before creation of pneumoperitoneum. Although the effect of bupivacaine peritoneal instillation is also noticeable when used after creation of pneumoperitoneum, it confers significantly lower benefits. PMID- 16703435 TI - Intrathoracoscopic localization techniques. Review of literature. AB - BACKGROUND: Several techniques for localizing pulmonary nodules have been described, but the advantages and disadvantages of each method remain unclear. We reviewed ultrasound, endofinger, finger palpation and wait and watch, radioguided, vital dye, fluoroscopic, agar marking, and needle wire methods for localizing pulmonary nodules. METHODS: Original, peer-reviewed, and full-length articles in English were searched with PubMed and ISI Web of Sciences. Case reports and case series with less than 10 patients were excluded. RESULTS: All localization techniques showed good reliability, but some carry a high rate of major or minor complications and drawbacks. CONCLUSION: No ideal localization technique is available; thus, the choice still depends on surgeon's preference and local availability of both specialists and instruments. PMID- 16703436 TI - Malignant gastrointestinal obstruction: endoscopic stenting versus surgical palliation. AB - BACKGROUND: Malignant gastrointestinal obstruction is a secondary complication of cancers in an advanced state. Treatment has consisted of gastrojejunostomy. However, the endoscopic placement of metallic stents has provided positive results. This study aimed to compare the efficiency of both therapeutic options. METHODS: A total of 41 patients with gastrointestinal obstruction caused by inoperable neoplasm were treated endoscopically with enteral stent (24 patients) or gastrojejunostomy (17 patients). RESULTS: In the endoscopic group (EG) 24 patients (100%) achieved efficient gastric emptying, as compared with 82.3% in the surgical group (SG). The difference was not significant. The average time for initiating oral food tolerance was 2.4 days for the EG and 5 days for the SG (p < 0.001). The average inpatient time was 7.1 days for the EG and 11.5 days for the SG (p < 0.001). Mortality at 30 days was lower in the EG (16.6%) than in the SG (29.4%) (p < 0.05). The survival time was 20 weeks for the EG and 21.6 weeks for the SG. The difference was not significant. The rate of complications was 4% in the (EG) and 17.6% in the (SG), with the difference was not significant. CONCLUSION: Endoscopic treatment of malignant gastrointestinal obstruction provides an adequate palliation of the symptoms. It is less invasive, avoids the morbidity associated with open gastrojejunostomy, and achieves a faster start to oral food and a shorter hospital stay, leading to a higher quality of life. PMID- 16703437 TI - Endoscopic treatment of acute colorectal obstruction with self-expandable metallic stents: experience in a community hospital. AB - BACKGROUND: The purpose of this study was to evaluate the efficacy and security of treatment with self-expandable metallic stents for acute colonic obstruction. METHODS: Retrospectively, we analyzed our experience in placing colonic stents in patients with acute colonic obstruction. Sixty-two patients with malignant colon obstruction were sent to our hospital endoscopic unit from February 1999 through September 2003. The mean age was 75.21 years. A total of 63 self-expandable metallic stents were implanted. All procedures were done under endoscopic and fluoroscopic guidance. RESULTS: Technical success was obtained in 58 patients (93.54%). Clinical improvement and resolution of the obstruction were confirmed in 56 of these 58 patients (96.55%) within 48-72 h. Sixteen complications were observed in 11 patients (19%). Twelve cases were minor complications: five patients had pain and rectal tenesmus, and there were seven cases of distal migration of the stent. Four patients (6.9%) had severe complications. Three patients underwent surgery to resolve colonic perforations and one patient developed a colovesical fistula 7 months after stent placement. The stent was used as a bridge to the elective surgery in 22 patients (37.93%). The mean time between stent placement and surgery was 7.66 days (range, 2-20). The stent was used as a definitive palliative treatment in 36 patients (62.07%). CONCLUSION: Stenting was a useful treatment of acute malignant colonic obstruction. The use of stents as a "bridge to the elective surgery" allowed the intestinal preparation, general status restoration, and a one-stage operation with resection and primary reanastomosis. We have also used the stents as a definitive palliative treatment, avoiding surgery in those patients with a very widely metastatic disease or who cannot undergo operation because of comorbid underlying conditions. PMID- 16703438 TI - Laparoscopic Heller myotomy for achalasia facilitated by robotic assistance. AB - BACKGROUND: Laparoscopic Heller myotomy is the standard operation for achalasia. The incidence of esophageal perforation is approximately 5% to 10%. Data about the safety and utility of robotically assisted Heller myotomy (RAHM) are scarce. The aim of this study was to assess the efficacy and safety of RAHM for the treatment of esophageal achalasia. METHODS: From a prospectively maintained database, demographic data, symptoms, esophagograms, manometries, and perioperative data from all the RAHMs performed between September 2002 and February 2004 were analyzed. RESULTS: A total of 54 patients underwent RAHM, including 26 men. The mean age of these patients was 43 years (range, 14-75 years). Dysphagia was present in 100% of the patients. Of the 54 patients, 26 (48%) had undergone previous treatment including pneumatic dilation (17 patients), Botox injections (4 patients), or both of these treatments (5 patients). The dissection was performed laparoscopically, and the myotomy was performed with robotic assistance. The operative time, including the robot setup time, averaged 162 min (range, 62-210 min). Blood loss averaged 24 ml. No mucosal perforations were observed. The hospital length of stay was 1.5 days. There were no deaths. At 17 months, 93% of the patients had relief of their dysphagia. CONCLUSIONS: The findings showed RAHM to be safe and effective, with a 0% incidence of perforation and relief of symptoms for 91% of the patients. PMID- 16703439 TI - Pouch enlargement and band slippage: two different entities. AB - BACKGROUND: Pouch enlargement and band slippage are the most common late complications of laparoscopic adjustable gastric banding (LAGB). Often, confusion exists among surgeons regarding the denomination or even the treatment of these two different entities. This study aimed to establish the differences in clinical presentation, radiologic features, and management between pouch enlargement and band slippage. The authors hypothesized that pouch enlargement can be managed nonoperatively (via band deflation), that band slippage is an acute complication requiring surgical treatment, and that tailored adjustment allows earlier diagnosis of pouch enlargement in asymptomatic patients. METHODS: From March 2001 to December 2004, 516 patients underwent LAGB placement. Barium swallow was performed preoperatively, postoperatively, and during band adjustments ("tailored adjustment"). Pouch enlargement was defined as dilation of the pouch, and band slippage was considered when band and stomach were prolapsed. Four radiologic types of pouch enlargement were considered: band 45 degrees, band 45 degrees with covering of the band, band 0 degrees, and band smaller than 0 degrees. RESULTS: A total of 1,600 barium swallows were performed with 516 patients. As a result, pouch enlargement was diagnosed for 61 patients (12%) and band slippage for 12 patients (2%). CONCLUSION: In this study, pouch enlargement was found to be a chronic complication that can be managed conservatively with a 77% success rate. Tailored adjustment allows early diagnosis of pouch enlargement, thus preventing adjustments in patients with undiagnosed pouch enlargement. Surgical treatment should be considered when medical treatment fails. By comparison, band slippage is an acute complication that requires surgical treatment in every case (100%). PMID- 16703440 TI - Laparoscopic rectal prolapse surgery combined with short hospital stay is safe in elderly and debilitated patients. AB - BACKGROUND: We report the results of patients treated from January 2000 to June 2004 for full-thickness rectal prolapse with trans-abdominal surgery in Helsinki. METHODS: Sixty-five of 75 patients were treated laparoscopically, with a 6% conversion rate. Ten patients were operated on openly. Half of the patients were scored as American Society for Anesthesiologists III or IV. RESULTS: The operation time was similar in the laparoscopic and the open rectopexy procedures (p = 0.15), whereas laparoscopic resection rectopexy was more time-consuming compared to the open procedure (p = 0.007). Intraoperative bleeding during laparoscopic surgery was minimal in comparison to open surgery (p = 0.006). Patients treated laparoscopically had a shorter median hospital stay than those treated with an open procedure (rectopexy, 3 and 7 days, respectively; resection rectopexy, 4 and 7.5 days, respectively) (p < 0.00001). There was no mortality and minor morbidity. During follow-up, there were two prolapse recurrences. All surgical techniques improved fecal continence considerably. Eighty-four percent of rectopexy patients and 92% of resection rectopexy patients considered the surgical outcome to be excellent or good. CONCLUSIONS: Both rectopexy and resection rectopexy cure prolapse with good results and can be performed safely in older and debilitated patients. The laparoscopic approach enables a shortened hospital stay and is well tolerated in elderly patients. PMID- 16703441 TI - Open versus laparoscopic appendicectomy: a critical review. AB - BACKGROUND: The benefit of laparoscopic appendicectomy remains unclear. We have analysed available randomised studies comparing laparoscopic and open appendicectomy regarding their clinical pitfalls and statistical relevance. METHODS: Thirty eight studies were analysed in terms of the following aspects: A. clinical problems (e.g., expertise of the surgeons, pre- and postoperative antibiotic treatment, definition of complications, blinding of outcomes) and B. statistical problems (e.g., definition of primary and secondary outcomes, power and sample size, statistical methods, confidence intervals, comparability of groups and studies). RESULTS: Most of the studies have clinical and statistical pitfalls. The most important pitfalls are the uncertain expertise of the operating surgeons, blinding, and the exploratory nature of the studies. Our analysis aims at giving useful information for the appraisal of existing studies and the conduct of further studies. It also gives some preliminary results. CONCLUSIONS: More than twenty years after Semm performed the first laparoscopic appendicectomy, it is necessary to clarify the superiority of laparoscopic or open appendectomy with well-defined, carefully designed randomised studies. PMID- 16703443 TI - NIS vs SAGES: a comparison of national and voluntary databases. AB - BACKGROUND: Surgical outcomes are increasingly examined in an effort to improve quality and reduce medical error. The Nationwide Inpatient Sample (NIS) is a retrospective, claims-derived and population-based database and the Society of American Gastrointestinal Endoscopic Surgeons (SAGES) Outcomes Project is a prospective, voluntary and specialty surgeon database. We hypothesized that these two sources of outcome data would differ in regard to a single, commonly performed procedure. METHODS: Both the NIS, a national sample of all nonfederal hospital discharges, and the gastroesophageal reflux disease log of the SAGES Outcomes Project were queried for all fundoplications performed between 1999 and 2001 using either ICD-9 procedure code 44.66 or CPT codes 43280 or 43324. Patients with an emergency admission, age <17 years, and/or diagnoses for either esophageal cancer or achalasia were excluded. Both demographic and outcome variables were compared by either t-test or chi-square analysis, with a p value of <0.05 as significant. RESULTS: Both data sets were comparable for age and gender; however, the SAGES group had a higher rate of teaching hospital affiliation (71 vs 48%, p < 0.001). SAGES fundoplications had a consistently higher rate of comorbidities, including Barrett's esophagus (2.3 vs 1.1%, p = 0.005). The NIS fundoplications had a clear trend toward more associated procedures, including cholecystectomy (7.2 vs 2%, p < 0.001). Complication rates for the NIS data set were higher, including pulmonary complications (1.7 vs 0.5%, p = 0.03). No statistically significant differences existed between the two data sets for either length of stay or mortality. CONCLUSIONS: The two databases indicate that fundoplication is an operation with low morbidity and mortality. The SAGES Outcomes Project demonstrated that participating surgeons had a higher affiliation with teaching hospitals, higher reporting of comorbidity, and lower associated procedures than the NIS. Despite having more comorbidity and technical difficulty, patients from the SAGES Outcomes Project had equivalent or lower complication rates. PMID- 16703442 TI - Inflammatory response and bacterial dissemination after laparotomy and abdominal CO2 insufflation in a murine model of peritonitis. AB - BACKGROUND: The immunologic repercussions due to cavity insufflation are the focus of great discussion. The aim of this study was to compare the inflammatory response and bacterial dissemination after laparotomy and abdominal CO2 insufflation in a murine model of peritonitis. METHODS: Swiss mice were inoculated intraperitoneally with 0.5 ml of a solution containing 1 x 10(8) colony-forming units (CFU)/ml of Escherichia coli and were divided into three groups as follow: control (anesthesia for 30 min), laparotomy (2.5-cm midline incision for 30 min), and CO2 pneumoperitoneum (CO2 cavity insufflation for 30 min). The number of leukocytes, CFU/ml counting, and the levels of interleukin (IL)-6, tumor necrosis factor-alpha (TNF-alpha), and IL-10 were evaluated in blood, peritoneal, and pleural fluid samples obtained at 90 min and 18 h after the procedures. RESULTS: The laparotomy group showed a greater bacterial dissemination to the blood, peritoneum, and pleural cavity and also greater neutrophil migration to the peritoneal cavity compared to the CO2 insufflated and control groups. The 24-h mortality was also significantly higher in the laparotomy group. The IL-6 levels showed a precocious rise in all groups submitted to bacterial inoculation at the 90-min time point. At the 18-h time point, IL-6 levels in the peritoneum were significantly higher in the laparotomy group than in the control or CO2 insufflated groups. At the same time, TNF-alpha levels were higher in the laparotomy and CO2 insufflated groups than in controls; IL-10 levels showed no differences among the groups. CONCLUSIONS: Our results suggest that cavity insufflation with CO2 is a more effective method of access, inducing less bacterial dissemination and also a less intense inflammatory response. Cavity insufflation with CO2 may present a good option for the surgical treatment of patients with bacterial peritonitis. PMID- 16703444 TI - The effect of taste and palatability on lingual swallowing pressure. AB - There is evidence that a strong, unpalatable, sour bolus improves swallowing in neurogenic dysphagia. It is not known whether other tastes may alter swallowing physiology. This study investigated the effect of moderate versus high taste concentrations (sweet, sour, salty, bitter) and barium taste samples on lingual swallowing pressure in ten healthy young adults, using a three-bulb lingual pressure array secured to the hard palate. Palatability of the samples was analyzed using the nine-point hedonic scale. Results showed that moderate sucrose, high salt, and high citric acid elicited significantly higher lingual swallowing pressures compared with the pressures generated by water. Pressures in the anterior bulb were significantly higher than those recorded from the middle or posterior bulb. There was no significant effect of palatability on lingual swallowing pressures. High salt and citric acid are known to elicit chemesthesis mediated by the trigeminal nerve. These results suggest that chemesthesis may play a crucial role in swallowing physiology. If true, dysphagia diet recommendations that include trigeminal irritants such as carbonation may be beneficial to individuals with dysphagia. However, before this recommendation more research is needed to examine how food properties and their perception affect swallowing in individuals with and without dysphagia. PMID- 16703445 TI - The challenge of mastication: preparing a bolus suitable for deglutition. AB - The main function of mastication is to transform a solid food into a bolus that can be swallowed safely. The bolus characteristics such as particles size or cohesiveness, are continuously sensed during mastication and they are important in initiating deglutition. This study examined the following question: What is the condition of the bolus just before swallowing? Ten subjects with normal dentition aged 37.5 +/- 3.7 years were asked to chew without swallowing six different foods (three nuts and three vegetables) while the number of cycles and the duration of the sequence were recorded. The particle size distribution shown by the expectorated food bolus just before swallowing was examined by image analysis. The results showed that, for a given food, the sizes of the bolus particles just before swallowing were comparable in all subjects. However, the number of cycles and duration of the sequence varied between subjects. Taken together these data strongly suggest that the granularity of the bolus before swallowing has to reach a predetermined state which is obtained by using an individual chewing strategy. This suggests that the bolus structure reflects a key factor for homeostasis and explains the large interindividual variability of the mastication physiologic parameters. PMID- 16703448 TI - Antinociceptive effect of botulinum toxin: an added value to chemical sphincterotomy in anal fissure? PMID- 16703449 TI - Increased experience and surgical technique lead to improved outcome after ileal pouch-anal anastomosis: a population-based study. AB - PURPOSE: This study was designed to determine whether changes in length of stay and 30-day readmission, reoperation, and excision rates for the ileal pouch-anal anastomosis occurred over time and with changes in surgical technique and hospital volume. METHODS: Using three population-based administrative databases, data on all ileal pouch-anal anastomoses performed in the province of Ontario between January 1992 and June 1998 were obtained. The effect of age, gender, stage of the procedure, year of surgery, and hospital volume were examined for their effect on length of stay and readmission, reoperation, and excision rates. RESULTS: There were 1,285 ileal pouch-anal anastomoses performed in 58 hospitals. There was a significant decrease in length of stay and reoperation and excision rates but a concommitant increase in readmission rate during the study period. Patients younger than aged 40 years had a significantly lower length of stay and excision rate. Patients who had a two-stage procedure had a shorter length of stay, readmission, and reoperative rate compared with those having a three-stage procedure. Hospital volume was a significant predictor of need for reoperation and excision with both low-volume and medium-volume hospitals having significantly higher rates than high-volume hospitals. CONCLUSIONS: Outcome after ileal pouch-anal anastomosis has improved. It is significantly better in patients younger than aged 40 years, having a two-stage procedure, and where surgery is performed at high-volume hospitals. It is likely that both modifications in surgical technique and surgical experience have led to improvements in clinical outcome after ileal pouch-anal anastomosis. PMID- 16703447 TI - Picoeukaryotic plankton diversity at the Helgoland time series site as assessed by three molecular methods. AB - We analyzed picoeukaryote assemblages in the German Bight at the Helgoland time series site by sequencing cloned eukaryotic 18S rRNA genes in six genetic libraries plus one library from the Orkney Islands from a cruise of opportunity. The libraries were constructed from environmental samples collected at different periods of the year. The same samples were also analyzed using a fingerprinting technique, single-strand conformational polymorphism (SSCP), and DNA microarrays with class-level oligonucleotide probes. One hundred unique clones were analyzed from each library, thus insuring over 85% coverage of the library. The V4 region of the 18S rRNA gene was sequenced from each of these clones, thus providing the most discrimination among the clones. The nonphotosynthetic picoeukaryotic component dominated over the photosynthetic one and was represented by the ciliates at 45% and group II alveolates at 42%. Prasinophytes dominated the photosynthetic group at 40%, but other picoplankton groups, such as bolidomonads and chrysophytes, were also present. Totally novel groups were found in the cryptomonads and in the dinoflagellates. A new algal group sister to the cryptophyte nuclear gene and the glaucocystophytes was also found. These three groups have been found in other picoeukaryotic planktonic clone libraries. SSCP analyses at closer time intervals suggest that clone libraries should be made at weekly intervals if succession in the picoeukaryotic plankton community is to be monitored accurately. A comparison of annual samples suggests thatthere appears to be an annual cycle with regard to species composition. Microarray analysis supported the clone library data and offered a faster means of community analysis, which can be performed with similar accuracy and with higher throughput for a more in-depth analysis. PMID- 16703451 TI - Assessing the impact of screening mammography: Breast cancer incidence and mortality rates in Connecticut (1943-2002). AB - BACKGROUND: Randomized controlled studies demonstrate that early detection and intervention reduce breast cancer mortality by approximately 25%. Though the ultimate goal of screening is to reduce breast cancer deaths, the immediate goal is to detect and treat early-stage tumors before they pose a threat to life. MATERIALS AND METHODS: To assess the impact of early detection and intervention in the general population, we analyzed breast cancer incidence and mortality rates in the NCI's Historical Connecticut Tumor Registry (1943-2002). RESULTS: Though breast cancer rates increased for the entire study period, overall incidence rates rose faster than previously following the initiation of mammography screening in the early 1980s in the United States. Of note, stage specific incidence rates increased 152% (53.2-133.9 per 100,000 woman-years) for early-stage tumors and fell 16% (56.1-47.2 per 100,000 woman-years) for late stage breast cancers. Period- and cohort-age-specific incidence rates rose dramatically for early-stage tumors among women targeted for screening (ages 40 80 years), whereas rates for regional and distant stages declined modestly among women ages >50 years. Breast cancer mortality rates fell 31.6%. CONCLUSIONS: Along with increases in incidence rates for early-stage tumors, rates for late stage disease and breast cancer mortality declined following widespread screening mammography, consistent with effective early detection and improved treatment over time. However, the disparity between the dramatic rise in early-stage tumors compared to the more modest declines in late-stage disease and mortality suggests that many mammography-derived early-stage lesions may never progress to late stage cancers and pose a threat to life. PMID- 16703452 TI - Papillary glioneuronal tumor. Case report and literature review. AB - OBJECTIVE AND IMPORTANCE: Papillary glioneuronal tumor is a recently described neoplasm composed of gliovascular pseudopapillae associated with intervening neuronal cells ranging from neurocytes to ganglion cells. This tumor is not currently included in the WHO classification of tumors of the central nervous system. We describe a new case of papillary glioneuronal tumor and analyze the data for a series of further 15 patients from international literature. CLINICAL PRESENTATION: A 27-year-old man presented to us for generalized seizure. CT and MRI showed a cystic tumor with mural nodule in the left frontal lobe. INTERVENTION: Frontal craniotomy with gross total removal of the tumor was performed. Histopathological examination was positive for papillary glioneuronal tumor. CONCLUSION: The clinical, radiologic, and pathological features of our case are strikingly similar to those of the previous reported cases. A review of the literature disclosed only 15 other cases of these tumors. It is important that every new case of PGNT is reported to allow its recognition and classification. PMID- 16703453 TI - Female predominance in meningiomas can not be explained by differences in progesterone, estrogen, or androgen receptor expression. AB - The female predominance in meningioma incidence and association between meningioma and breast cancer suggest that growth of meningiomas is hormone dependent. There are several discrepancies in literature about the proliferative effect of sex hormones on meningiomas. This study aims to evaluate the hormone receptor status of meningiomas and assess its relation to age, sex, histological grade, recurrence, and proliferation activity. The material was based on consecutive patients operated for meningioma at Tampere University Hospital in 1989-1999. The occurrence of progesterone, estrogen and androgen receptor in patients with primary and recurrent meningiomas was studied immunohistochemically by using specific monoclonal antibodies. Hormonal status was determined in 510 tumor samples. 443 samples were from primary meningiomas and 67 from recurrent tumors. Of the samples, 455 were benign (WHO grade I), 49 atypical (grade II), and 6 malignant (grade III). Of the primary tumor samples, 88% were progesterone receptor positive, 40% were positive for estrogen and 39% for androgen receptors. Grade I meningiomas had significantly higher incidence for estrogen and androgen receptors than higher grade meningiomas. Estrogen positive tumor samples had significantly higher proliferation index than estrogen negative samples. No difference in expression of sex hormone receptors was observed by sexes or age group. Estrogen and androgen receptors may have more influence on the pathogenesis of meningiomas than earlier thought. The higher incidence of meningiomas in women can not be explained by differences of sex hormone receptor expression. PMID- 16703455 TI - Chemical and biological evaluation of sediments from the Wadden Sea, The Netherlands. AB - We describe the results of an evaluation of marine sediments using chemical measurements and bioassays. Four groups of chemicals, i.e., heavy metals, PAHs, chlorinated aromatic compounds, and tin compounds, were measured at 16 locations in the Wadden Sea, The Netherlands. Extractions of sediments from each location also were assessed using five bioassays. Our objective was to identify chemicals likely to pose biological risks, characterize the relation between bioassay results and particular classes of chemical(s) and determine "clean" reference sites on the basis of the chemical and biological evaluations. A multivariate technique (Principal Component Analysis; PCA) was used to meet these objectives. Results of the PCA indicated that the response of the Microtox Solid Phase bioassay had a positive, significant relationship with the levels of PAHs and organotin compounds. The responses of the other bioassays did not consistently relate to the concentrations of the other measured chemicals. Our findings indicate that the organotin compounds may still be a stressor for aquatic invertebrates in the Dutch Wadden Sea. On the basis of the chemical and biological evaluations, four sites (Dantziggat, Malzwin, Richel and Lauwers) can be considered to be "clean" reference sites. PMID- 16703454 TI - The potential for phytoremediation of iron cyanide complex by willows. AB - Hybrid willows (Salix matsudana Koidz x Salix alba L.), weeping willows (Salix babylonica L.) and hankow willows (Salix matsudana Koidz) were exposed to potassium ferrocyanide to determine the potential of these plants to extract, transport and metabolize this iron cyanide complex. Young rooted cuttings were grown in hydroponic solution at 24.0 +/- 0.5 degrees C for 144 h. Ferrocyanide in solution, air, and aerial tissues of plants was analyzed spectrophotometrically. Uptake of ferrocyanide from the aqueous solution by plants was evident for all treatments and varied with plant species, ranging from 8.64 to 15.67% of initial mass. The uptake processes observed from hydroponic solution showed exponential disappearance kinetics. Very little amounts of the applied ferrocyanide were detected in all parts of plant materials, confirming passage of ferrocyanide through the plants. No ferrocyanide in air was found due to plant transpiration. Mass balance analysis showed that a large fraction of the reduction of initial mass in hydroponic solution was metabolized during transport within the plant materials. The difference in the metabolic rate of ferrocyanide between the three plant species was comparably small, indicating transport of ferrocyanide from hydroponic solution to plant materials and further transport within plant materials was a limiting step for assimilating this iron cyanide complex. In conclusion, phytoremediation of ferrocyanide by the plants tested in this study has potential field application. PMID- 16703456 TI - Biomarkers of environmental contaminants in field population of green mussel (Perna viridis) from Karnataka-Kerala coast (South West coast of India). AB - The green mussel Perna viridis was sampled from relatively clean and contaminated sites along the Kartanata-Kerala coast (south west coast of India) to study the tissue concentration of trace metals and biological responses to stress (biomarkers) such as sister chromatid exchange (SCE), chromosomal aberration, micronucleus (MN) test, hemic neoplasia (HN), Chromotest (Ames test) and comet assay. In general, mean tissue concentrations of toxic trace metals collected from 25 sampling sites were found to be below the World Health Organisation (WHO) permissible concentration given for seafood. The digestive gland extract of mussels from all 25 sampling sites showed negative reaction for mutagenic activity (Ames test) in the absence of metabolic activation. Very low levels of chromosomal aberration, SCE, MN, HN and comet cells were observed in mussels collected from both the urban associated and relatively clean sites. This study seems to indicate that that the coastal waters of Karnataka and Kerala are minimally contaminated with genotoxic and carcinogenic chemicals. PMID- 16703457 TI - Metal and antibiotic-resistance in psychrotrophic bacteria from Antarctic Marine waters. AB - In the wake of the findings that Antarctic krills concentrate heavy metals at ppm level, (Yamamoto et al. 1987), the Antarctic waters from the Indian side were examined for the incidence of metal and antibiotic-resistant bacteria during the austral summer (13th Indian Antarctic expedition) along the cruise track extending from 50 degrees S and 18 degrees E to 65 degrees S and 30 degrees E. The bacterial isolates from these waters showed varying degrees of resistance to antibiotics (Chloramphenicol, ampicillin, streptomycin, tetracycline and kanamycin) and metals (K(2)CrO(4), CdCl(2), ZnCl(2) and HgCl(2)) tested. Of the isolates screened, about 29% and 16% were resistant to 100 ppm of cadmium and chromium salt respectively. Tolerance to lower concentration (10 ppm) of mercury (Hg) was observed in 68% of the isolates. Depending on the antibiotics the isolates showed different percentage of resistance. Multiple drug and metal resistance were observed. High incidence of resistance to both antibiotics and metals were common among the pigmented bacterial isolates. Increased resistance decreased the ability of bacteria to express enzymes. The results reiterate previous findings by other researchers that the waters of southern ocean may not be exempt from the spread of metal and antibiotic-resistance. PMID- 16703468 TI - Mutagenesis studies on the N-terminus and Thr54 of Naja naja atra (Taiwan cobra) chymotrypsin inhibitor. AB - Ala-screening mutagenesis studies on Arg1, Pro2, Arg3, Phe4 and Thr54 of Naja naja atra (Taiwan cobra) chymotrypsin inhibitor showed that inhibitory potency and gross conformation of the mutants were not significantly different from those of wild-type inhibitor. Nevertheless, the R1A mutant had an appreciable decrease in the structural stability underlying thermal unfolding and urea-induced denaturation. Alternatively, deleting the first three residues at the N-terminus caused a reduction in structural stability as well as inhibitory potency. In sharp contrast to wild-type and other mutated inhibitors, R1A mutant and truncated mutant completely lost their inhibitory activity when the inhibitors were incubated with chymotrypsin for periods of up to 3 h. The loss of activity correlated with chymotryptic cleavage of inhibitors as evidenced by SDA-PAGE. Taken together, these results reflect that the globally structural rigidity of N. naja atra chymotrypsin inhibitor functionally affects the sustainable period in inhibiting chymotrypsin activity, and that the intact N-terminus might contribute to this event. PMID- 16703458 TI - Evaluating the effect of sulphated polysaccharides on cyclosporine a induced oxidative renal injury. AB - Cyclosporine A (CsA) has been universally used as an immunosuppressant for the management of organ transplantation and various autoimmune diseases. However, nephrotoxicity due to CsA remains to be an important clinical challenge. In the present investigation, an attempt has been made to appraise the effect of sulphated polysaccharides on oxidative renal injury caused by CsA. Adult male Wistar rats were divided into four groups. Two groups received CsA by oral gavage (25 mg/kg body weight) for 21 days to provoke nephrotoxicity, one of which simultaneously received sulphated polysaccharides subcutaneously, (5 mg/kg body weight). A vehicle (olive oil) treated control group and sulphated polysaccharides drug control were also built-in. An increase in lipid peroxidation along with abnormal levels of enzymic (superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione-S transferase and glucose-6-phosphate dehydrogenase) and non-enzymic antioxidants (glutathione, vitamin C and vitamin E) are the salient features observed in CsA induced nephrotoxicity. CsA induced impairment of renal toxicity was evident from the marked decline in the activities of renal marker enzymes like alkaline phosphatase, acid phosphatase and lactate dehydrogenase, as well as an apparent increase in the serum urea, uric acid and creatinine; diagnostic of renal damage was normalized by sulphated polysaccharides co-administration. Sulphated polysaccharides treatment showed an effectual role in counteracting the free radical toxicity by bringing about a significant decrease in peroxidative levels and increase in antioxidant status. These observations emphasize the antioxidant property of sulphated polysaccharides and its cytoprotective action against CsA induced nephrotoxicity. PMID- 16703469 TI - Functional contribution of Ca2+ and Mg2+ to the intermolecular interaction of visinin-like proteins. AB - The interaction of human visinin-like protein 1 (VILIP1) and visinin-like protein 3 (VILIP3) with divalent cations (Mg2+, Ca2+, Sr2+ and Ba2+) was explored using circular dichroism and fluorescence measurement. These results showed that the four cations each induced a different subtle change in the conformation of VILIPs. Moreover, VILIP1 and VILIP3 bound with Ca2+ or Mg2+ in a cooperative manner. Studies on the truncated mutants showed that the intact EF-3 and EF-4 were essential for the binding of VILIP1 with Ca2+ and Mg2+. Pull-down assay revealed that Ca2+ and Mg2+ enhanced the intermolecular interaction of VILIPs, and led to the formation of homo- and hetero-oligomer of VILIPs. Together with previous findings that Ca2+-dependent localization of VILIPs may be involved in the regulation of distinct cascades and deprivation of Ca2+-binding capacity of VILIPs did not completely eliminate their activity, it is likely to reflect that Mg2+-bound VILIPs may play a role in regulating the biological function of VILIPs in response to a concentration fluctuation of Ca2+ in cells. PMID- 16703470 TI - Prediction of mitochondrial proteins using discrete wavelet transform. AB - A new method was proposed for prediction of mitochondrial proteins by the discrete wavelet transform, based on the sequence-scale similarity measurement. This sequence-scale similarity, revealing more information than other conventional methods, does not rely on subcellular location information and can directly predict protein sequences with different length. In our experiments, 499 mitochondrial protein sequences, constituting a mitochondria database, were used as training dataset, and 681 non-mitochondrial protein sequences were tested. The system can predict these sequences with sensitivity, specificity, accuracy and MCC of 50.30%, 95.74%, 76.53% and 0.54, respectively. Source code of the new program is available on request from the authors. PMID- 16703471 TI - Identification of glutamate residues important for catalytic activity or thermostability of a truncated Bacillus sp. strain TS-23 alpha-amylase by site directed mutagenesis. AB - The importance of 17 glutamate residues of a truncated Bacillus sp. strain TS-23 alpha-amylase (BACdeltaNC) was investigated by site-directed mutagenesis. The Ala and Asp-substituted variants were overexpressed in the recombinant E. coli cells and the 54-kDa proteins were purified to nearly homologous by nickel-chelate chromatography. Glu-295, which locates in the conserved region III of amylolytic enzymes, mutations resulted in a complete loss of enzyme activity. The specific activity for E151A was decreased by more than 30%, while other variants showed activity comparable to that of BACdeltaNC. A decreased half-life at 70 degrees C was observed for Glu-219 variants with respective to the wild-type enzyme, suggesting that replacement of Glu-219 by either Ala or Asp might have a significant destabilizing effect on the protein structure. PMID- 16703472 TI - Purification, characterization and cloning of phospholipase D from peanut seeds. AB - We purified phospholipase D (PLD) enzyme from peanut seeds, and the PLD enzyme eluted as two distinct peak fractions on Mono-Q chromatography, the first of which was characterized. N-terminal sequencing indicated that the N-terminus was blocked. The molecular mass of the purified enzyme was estimated to be 92 kDa by SDS-PAGE. The pH optimum of the enzyme was 5.0, and the Km value against its substrate phosphatidylcholine (PC), in the presence of 10 mM CaCl2 and 4 mM deoxycholate, was estimated to be 0.072 mM. The enzyme catalyzed two reactions, i.e., hydrolysis of PC generating phosphatidic acid (PA) and choline, and transphosphatidylation of the PA-moiety in the PC molecule to the acceptor glycerol, generating phosphatidylglycerol. Furthermore, we cloned two types of full-length cDNA, Ahpld1 and Ahpld2, each encoding distinct PLD molecules having 794 and 807 residues, respectively. The partial amino acid sequence of the purified PLD was consistent with the deduced sequence of AhPLD2. PMID- 16703473 TI - Psychometrics of the recovery process inventory. AB - The reliability and validity of the Recovery Process Inventory (RPI) were assessed in a statewide sample of 459 persons with severe mental illness from a public mental health system. Six factors were identified that reflect important aspects of the recovery process, have good internal consistency and fair to moderate test-retest reliability, and yield moderate to good convergent validity on the majority of subscales. The 22-item RPI performance across these dimensions and its comparability to the Recovery Assessment Scale and related instruments, e.g., empowerment, confidence, and healing, make it a potentially valuable tool in survey research regarding the process and outcomes of recovery among mental health consumers. Suggestions for its further research and development, and use of the scale in conjunction with the Mental Health Statistics Improvement Program Adult Consumer Survey are discussed. PMID- 16703481 TI - [Endobronchial tumor presenting as asthma]. PMID- 16703480 TI - [Comparison of patterns of metastasis between malignant pleural mesotheliomas and pulmonary carcinomas]. AB - Starting with the question whether there is a principal difference in the metastatic behaviour of pleural mesothelioma and pulmonary tumors, a detailed retrospective study of the findings of 210 post-mortem examinations between the years of 1992 to 1999 was conducted. The spectrum of metastatic sites is very large in malignant pulmonary carcinomas (n = 148) as well as in malignant pleural mesotheliomas (n = 62). There is no significant difference in the TMN-staging between both tumor groups at the time of death. One exception was the brain: there were significantly more metastases from the pulmonary tumors. Carcinomas metastasized more frequently into the skeleton, the kidneys and the adrenal glands, while metastases of mesotheliomas were more often found in the peritoneum. As of the moment there is no significant difference in the pattern of metastasis between both tumor groups discernible. PMID- 16703482 TI - [Swimming pool lung -- extrinsic allergic alveolitis or mycobacterial disease?]. AB - There have been several recent reports of pulmonary disease resulting from exposure to Mycobacterium avium complex in indoor hot tubs. The disease is thought to be due either to infection or extrinsic allergic alveolitis (EAA). In this report we describe the case of a patient who developed episodes of fever, dyspnea and cough 4-6 hours after cleaning his indoor swimming pool. A diagnosis of EAA was made on finding a restrictive lung function pattern with gas exchange abnormalities, a predominant lymphocytosis in the bronchoalveolar lavage, diffuse ground-glass opacities in the lower lobes on high-resolution computer tomography, and specific IgG antibody activity to the swimming pool water. There was no precipitin reaction or specific IgG antibody activity to microbes extracted from the water. Interestingly, the water contained Mycobacterium avium complex (MAC) in huge amounts and in this case the histopathological features of the lung biopsy specimens differed from those seen in typical EAA, but were similar to those described in "hot tub lung" caused by mycobacteria. Solely by avoidance of cleaning the swimming pool, without any pharmacological treatment, the patient recovered completely within three months. To the best of our knowledge, this is the first report of EAA possibly associated with MAC exposure in a swimming pool environment. PMID- 16703483 TI - [Felix Schumann]. PMID- 16703484 TI - [A new concept for the treatment of patients with acute exacerbations of chronic obstructive pulmonary disease (COPD)]. AB - In practice, antimicrobial treatment is usually thought to be regularly indicated in the treatment of patients with acute exacerbations of COPD. However, data supporting this practice are very limited. Most placebo-controlled studies do not satisfy currently required methodological quality standards. We develop a concept of restrictive indications for antimicrobial treatment from the available data. Cornerstones of this concept include the selection of treatment setting as well as the indication for antimicrobial treatment according to the severity of the acute exacerbation (not primarily of COPD) and to the type of exacerbation (Anthonisen type I). The selection of antimicrobial agents is based on an intraindividual rotation concept in order to limit antimicrobial selection pressure. The concept presented here can at the same time be used as a clinical guide as well as a framework for future studies dealing with currently unresolved issues. PMID- 16703485 TI - [Working place related obstructive airway diseases]. PMID- 16703486 TI - 3D power Doppler analysis of the vascularisation in tumours of the oral cavity. AB - PURPOSE: The aim of this study was to determine the volume and vascularisation of tumours by three-dimensional (3D) power Doppler ultrasound in patients with "tumours of the oral cavity". MATERIALS AND METHODS: Three-dimensional (3D) power Doppler ultrasound investigation was performed pre-operatively on 49 patients with carcinomas of the oral cavity. Digitally-stored data were analysed with 3D View program software, a part of which is the "VOCAL-Imaging program". By using VOCAL, the borders of a structure (tumour) can be determined and its volume calculated. The vascularisation of the tumour was determined by analysing three dimensional colour histograms, and the vascularisation indices were calculated. RESULTS: Vascularisation indices VI and VFI were significantly higher in patients with neck metastases. The differences between the vascularisation indices in N+ and N0 necks were statistically significant (p < 0.05). The flow index (FI) values between N+ and N0 necks were not statistically different. CONCLUSION: The volume and vascularisation of tumours of the oral cavity could be determined effectively by three-dimensional power Doppler ultrasound. Tumour vascularisation is higher in metastatic than in non-metastatic tumours. PMID- 16703487 TI - Ultrasound Doppler of the Achilles tendon before and after injection of an ultrasound contrast agent--findings in asymptomatic subjects. AB - The sensitivity of ultrasound Doppler has now reached a level at which perfusion can be detected even in normal, resting musculoskeletal tissues. To be able to distinguish normal from abnormal flow, the resistive index (RI) determined by spectral Doppler may be of value. Inflammation is associated with low RI values and a value of 1.00 is normal. PURPOSE: In tissues with no detectable flow, it has been assumed that RI may be defined as 1.00. This study was performed to test if normal tendinous vessels could be visualised with a contrast agent, and if such vessels had normal RI. MATERIALS AND METHODS: Ultrasound (US) was performed with a 14 MHz linear transducer. 22 asymptomatic tendons in 12 subjects were scanned, and only 5 ultrasonically normal tendons in 5 subjects were identified. In these 5 normal tendons, SonoVue was used to make vessels visible. The Achilles tendons and the peritendinous tissues were evaluated clinically. RESULTS: In all 5 ultrasonically normal tendons, arteries could be detected with Doppler after contrast injection and in all cases with normal RI values. All vessels were located in the mid-portion of the tendon. 18 tendons were categorised as normal by the clinician. CONCLUSION: After administration of a contrast agent, all Achilles tendons could be demonstrated to have arteries present in the mid portion. The flow profile of these arteries was fully normal without diastolic component and with an RI of 1.00. This finding has implications for criteria of normality of tendons, which cannot be based exclusively on the presence or absence of Doppler activity. The majority of the tendons in this study had ultrasound abnormalities both on gray-scale and colour Doppler. It can be speculated that the abnormalities may be age-related degenerative changes and that age-stratified normal materials are needed to define normality. There was no consistency between US and clinical diagnosis. PMID- 16703488 TI - The syndrome of right isomerism -- prenatal diagnosis and outcome. AB - OBJECTIVE: To evaluate the accuracy of the prenatal diagnosis of right isomerism and to assess possible diagnostic and prognostic markers. METHODS: Retrospective review of all cases of right isomerism identified between 1989 and 2003 in two tertiary referral centres in Germany. RESULTS: Among 21 foetuses, 16 had a correct prenatal diagnosis of right isomerism. 19 showed different types of viscerocardiac heterotaxy, 12 of them in combination with juxtaposition of vena cava inferior and aorta. 20 had cardiac defects, with a high prevalence of atrioventricular septal defect (CAVSD) (62 %), right outflow tract obstruction (48 %), anomalous pulmonary venous return (33 %) and double outlet right ventricle (29 %). 4 out of 6 cases with total anomalous pulmonary venous return were overseen on prenatal ultrasound. Only 6 children survived. The highest loss occurred in the neonatal period. Two out of 6 survivors underwent single ventricle palliation, while another two had a biventricular repair. One child is awaiting mitral valve replacement. The remaining case has no cardiac defect and lives with supraventricular re-entry tachycardia. Only the presence of CAVSD was significantly correlated with non-survival (p < 0.05). CONCLUSIONS: The prenatal diagnosis of right isomerism remains a difficult task. Important sonographic markers are viscerocardiac heterotaxy, complex cardiac malformations and juxtaposition of vena cava inferior and aorta. Special attention has to be paid to the pattern of pulmonary venous drainage, as it is often misdiagnosed. The mortality in neonates is high, especially in the presence of CAVSD. Survivors suffer from significant morbidity. PMID- 16703490 TI - Intimal flap in a common carotid artery in a patient with Marfan's syndrome. AB - Dissection of the common carotid artery is a rare but important complication of Marfan's syndrome. The following case describes a patient with formation of an intimal flap of the common carotid artery who had suffered from an aortic dissection years before. PMID- 16703489 TI - A rare case of massive ovarian edema in an uncomplicated singleton pregnancy. AB - Massive Ovarian Edema (MOE) is a rare cause of ovarian enlargement. Only 77 cases have been reported in the world literature so far, merely three of which were observed during pregnancy. Two of these showed additional pathological changes of the affected ovary (torsion) or were found in twin pregnancy. The patient presented on hand is therefore only the second report of MOE in an uncomplicated singleton pregnancy. In this case, a therapeutical resection of the affected ovary was performed at 16 + 4 weeks of gestation. The further course of the pregnancy was uncomplicated. PMID- 16703491 TI - [Liver cirrhosis as sequela of selective hepatic radio-embolization with Yttrium (Y-90) microspheres]. PMID- 16703492 TI - [Economical role of radiology in medical cooperation]. PMID- 16703493 TI - [Imaging of enthesitis: a new field for the radiologist?]. AB - Entheses are sites where tendons, ligaments, joint capsules, or fasciae attach to bone. Their function is to provide a mechanism for reducing stress at the bony interface by dissipating the biomechanical load acting on the bone. Enthesitis may occur in traumatic, endocrinologic, metabolic, degenerative, or inflammatory conditions and is a major symptom in patients with seronegative spondyloarthritis (SpA). The increasing interest in inflammation of the entheses associated with rheumatoid diseases has fundamentally changed our understanding of the clinical presentation, pathogenesis, and therapy of enthesitis. Conventional radiographs and computed tomography (CT) allow good evaluation of chronic changes of the entheses, such as soft-tissue calcification, erosions, and new bone formation. The method of first choice to evaluate acute enthesitis is magnetic resonance imaging (MRI), which depicts both soft-tissue changes and intraosseous abnormalities. The purpose of this overview is to discuss the MRI appearance of enthesitis in patients with SpA and to provide a morphologic survey of the predominantly affected entheses. PMID- 16703494 TI - [Magnetic resonance imaging of glenohumeral instability]. AB - Conventional MR imaging and MR arthrography are established diagnostic imaging modalities for investigating shoulder instability. Since there are currently various surgical shoulder stabilization methods as well as conservative treatment strategies, the role of imaging is to provide diagnostic information to help determine the therapeutic approach. Whereas conventional MR imaging is usually sufficient for the evaluation of acute shoulder injuries due to the presence of a posttraumatic joint effusion, MR arthrography is the imaging modality of choice for chronic shoulder instability. Atraumatic and microtraumatic instability of the shoulder must be distinguished from traumatic instability since clinical findings and secondary or associated injuries differ from those of traumatic instability. Injuries of the IGHL-complex can be reliably diagnosed with MR arthrography. Traumatic anteroinferior luxation causes labroligamentous injuries at the glenoid insertion (Bankart-, Perthes-, ALPSA-, and non-classifiable chronic lesions) and injuries of the IGHL and its humeral insertion (HAGL-, BHAGL , and floating AIGL-lesions). The type of injury and extent of degenerative changes or scarring that can be assessed with MR arthrography influence the therapeutic approach and in particular the decision between arthroscopic and open surgical methods of stabilization. PMID- 16703495 TI - Multislice CT coronary angiography: effect of sublingual nitroglycerine on the diameter of coronary arteries. AB - PURPOSE: To investigate the influence of sublingual glycerol trinitrate (1.2 mg, Nitrate [nitroglycerine], Nitrolingual N Spray) on the coronary artery diameter on multislice computed tomography (MSCT) coronary angiography. MATERIALS AND METHODS: Out of our database of patients who underwent MSCT (slice thickness of 0.5 mm, Aquilion, Toshiba) coronary angiography between July 2003 and November 2005 (950 patients) we retrospectively identified patients with follow-up examinations who received Nitrate for one examination while another examination was performed without Nitrate (10 patients). Another 10 patients who underwent two MSCT examinations with sublingual Nitrate administration were randomly selected from this database to serve as control group. For the resulting 40 MSCT examinations, blinded MSCT datasets were prepared, which were randomly evaluated by a reader blinded to the patient information and whether or not Nitrate had been given. The proximal coronary artery diameters were measured for the left main coronary artery (LMA), the left anterior descending coronary artery (LAD), the left circumflex coronary artery (LCX), and the right coronary artery (RCA) in all 40 datasets, resulting in altogether 160 measurements. RESULTS: The proximal diameters of all four coronary arteries were significantly larger on the MSCT coronary angiograms obtained after sublingual administration of Nitrate compared with the examinations in the same 10 patients without Nitrate (p < 0.001). The average diameters without and with Nitrate for the LMA, LAD, LCX, and RCA were 4.3 +/- 1.1 vs. 4.8 +/- 0.9 mm (12 % increase, p < 0.005), 3.0 +/- 0.6 vs. 3.5 +/ 0.5 mm (17 % increase, p < 0.001), 2.7 +/- 0.6 vs. 3.2 +/- 0.7 mm (19 % increase, p < 0.005), and 2.9 +/- 0.9 vs. 3.5 +/- 0.7 mm (21 % increase, p < 0.005), respectively. In the control group of 10 patients who underwent two MSCT coronary angiographies after sublingual Nitrate, no significant difference in the proximal diameter of all four main coronary vessels was observed. CONCLUSION: Sublingual administration of Nitrate results in significantly larger proximal coronary artery diameters on MSCT coronary angiography and might thus be recommended for routine examinations. PMID- 16703496 TI - [Technical Note: Subclavian artery misplacement of a 12F Shaldon catheter: percutaneous repair with a local closure device under temporary balloon tamponade]. AB - A case of subsequent percutaneous repair using a local closure device with a collagen block (VasoSeal(R)) and temporary balloon tamponade after inadvertent subclavian artery misplacement of a 12F Shaldon catheter is reported. Balloon occlusion safely prevented displacement of collagen into the vascular lumen by occluding the 12F entry site. Furthermore, collagen-induced coagulation was facilitated. PMID- 16703497 TI - [CT-guided vertebroplasty and kyphoplasty: comparing technical success rate and complications in 101 cases]. AB - PURPOSE: To compare the technical success and complication rates in CT-guided vertebroplasty and kyphoplasty. MATERIALS AND METHODS: From 2002 - 2005 69 patients (101 vertebrae) were treated with vertebroplasty (n = 82) or kyphoplasty (n = 19) using 4-slice MSCT with CT fluoroscopy as the sole guidance for the procedure. The underlying lesions were osteoporotic fractures in 78 vertebral bodies and 23 vertebral metastases. RESULTS: Technical success was achieved in all 101 procedures. Post-interventional CT demonstrated asymptomatic cement leakage in 51/101 vertebrae. Kyphoplasty resulted in leakage in 11/19 (57.9 %) and vertebroplasty in 40/82 (48.8 %) procedures. With p = 0.48 (Mann-Whitney Test) there was no significant difference between kyphoplasty and vertebroplasty with respect to the number of cement leakage occurrences. There was one minor complication of an L5 root irritation following radiofrequency ablation and vertebroplasty of a sarcoma metastasis which subsided without treatment after 8 weeks. There was one major complication of intraspinal cement leakage during tumor vertebroplasty causing T5 root compression and requiring laminectomy for cement removal. The overall rate of major complications requiring treatment was 0.99 %. CONCLUSION: Vertebroplasty and kyphoplasty can be safely performed using only MSCT fluoroscopy guidance. The rate of major complications is very low. There was a high rate of small asymptomatic cement leakages which may have remained undetected with conventional fluoroscopy (CF). There was no statistically significant advantage for kyphoplasty with respect to cement leakage and the technical success rate. PMID- 16703498 TI - [Cerebral lesions in acute arterial hypertension: the characteristic MRI in hypertensive encephalopathy]. AB - PURPOSE: In the nine years since the posterior reversible (leuc) encephalopathy syndrome (PRES) was first described, a number of causes have been under discussion. These not only include arterial hypertension, i. e. hypertensive crises, but also various toxic substances, i. e. immunosuppressive or chemotherapeutic agents, that are responsible for the formation of the symptoms and characteristic MR tomographic brain findings. MATERIALS AND METHODS: Initial and follow-up MRI examinations of 8 patients were analyzed. All patients had acute neurological symptoms (headaches, seizures, visual disorders and vigilance disturbances) together with a detectable hypertensive crisis. RESULTS: MRI disclosed increased signal intensity in subcortical and some cortical lesions in all patient FLAIR sequences. These changes were particularly extensive in the posterior circulation (occipital, cerebellum and brain stem) although they were also detected in brain areas supplied by the carotid artery. However, a cytotoxic genesis of the changes was ruled out in each patient by means of a normal DWI. Furthermore, when the blood pressure was normalized, reversibility of the lesions as proof of the diagnosis was detectable. CONCLUSION: The imaging findings can be typically analyzed as a predominantly posterior distribution of encephalopathic lesions with a high probability of reversibility after lowering blood pressure was patients suffering from a critical increase in blood pressure with corresponding neurological symptoms. The exact pathophysiology remains unclear, but the cause currently most favored is a vasculopathy of the posterior circulation due to diminished adrenergic autoregulation in combination with a dysfunction of the endothelial cells. In conclusion, we suggest designating this subpopulation from the non-uniform pool of patients with posterior (leuc) encephalopathy as "hypertensive encephalopathy". "Hypertensive encephalopathy" has to be distinguished from "toxic encephalopathy", particularly due to different therapeutic and prognostic consequences. PMID- 16703499 TI - [Application of (1)H MR spectroscopic imaging in radiation oncology: choline as a marker for determining the relative probability of tumor progression after radiation of glial brain tumors]. AB - PURPOSE: To determine the relative signal intensity ratios of choline (Cho), phosphocreatine (CR) and N-acetyl-aspartate (NAA) in MR spectroscopic imaging (proton-MRSI) for differentiating progressive tumors (PT) from non-progressive tumors (nPT) in follow-up and treatment planning of gliomas. Threshold values to indicate the probability of a progressive tumor were also calculated. MATERIAL AND METHODS: Thirty-four patients with histologically proven gliomas showing a suspicious brain lesion in MRI after stereotactic radiotherapy were evaluated on a 1.5 Tesla unit (Magnetom Vision, Siemens, Erlangen, Germany) using 2D proton MRSI (repetition time/echo time = 1500/135 msec, PRESS; voxel size 9 x 9 x 15 mm (3)). A total of 274 spectra were analyzed (92 voxel were localized within the suspicious brain lesion). Relative signal intensities Cho, Cr and NAA were measured and their ability to discern between PT and nPT was assessed using the linear discrimination method, logistic regression, and the cross-validation method. PT and nPT were differentiated between on the basis of clinical course and follow-up by MRI, CT and positron emission tomography. RESULTS: The Cho parameter and the relative signal intensity ratios of Cr and NAA were most effective in differentiating between PT and nPT. The logistic regression method using the parameter ln(Cho/Cr) and ln(Cho/NAA) had the best predictive results in cross-validation. A sensitivity of 93.8 % and specificity of 85.7 % were achieved in the differentiation of PT from nPT by proton-MRSI. CONCLUSION: (1)H-MRSI has a high sensitivity and specificity for differentiating between therapy-related effects and the relapse of irradiated gliomas. This method allows for assessment of the probability of radiotherapy response or failure. PMID- 16703500 TI - [Multifocal soft tissue tuberculosis in a patient without acute pulmonary involvement]. PMID- 16703501 TI - [Chronic hematogenous osteomyelitis in an unusual location]. PMID- 16703504 TI - Distribution of amantadine-resistant H5N1 avian influenza variants in Asia. AB - We examined the distribution of genetic mutations associated with resistance to the M2 ion channel-blocking adamantane derivatives, amantadine and rimantadine, among H5N1 viruses isolated in Vietnam, Thailand, Cambodia, Indonesia, Hong Kong, and China. More than 95% of the viruses isolated in Vietnam and Thailand contained resistance mutations, but resistant mutants were less commonly isolated in Indonesia (6.3% of isolates) and China (8.9% of isolates), where human infection was recently reported. The dual mutation motif Leu26Ile-Ser31Asn (leucine-->isoleucine at aa 26 and serine-->asparagine at aa 31) was found almost exclusively in all resistant isolates from Vietnam, Thailand, and Cambodia, suggesting the biological selection of these mutations. PMID- 16703503 TI - Airflow decline after myeloablative allogeneic hematopoietic cell transplantation: the role of community respiratory viruses. AB - We conducted a 12-year retrospective study to determine the effects that the community respiratory-virus species and the localization of respiratory-tract virus infection have on severe airflow decline, a serious and fatal complication occurring after hematopoietic cell transplantation (HCT). Of 132 HCT recipients with respiratory-tract virus infection during the initial 100 days after HCT, 50 (38%) developed airflow decline < or =1 year after HCT. Lower-respiratory-tract infection with parainfluenza (odds ratio [OR], 17.9 [95% confidence interval {CI}, 2.0-160]; P=.01) and respiratory syncytial virus (OR, 3.6 [95% CI, 1.0-13]; P=.05) independently increased the risk of development of airflow decline < or =1 year after HCT. The airflow decline was immediately detectable after infection and was strongest for lower-respiratory-tract infection with parainfluenza virus; it stabilized during the months after the respiratory-tract virus infection, but, at < or =1 year after HCT, the initial lung function was not restored. Thus, community respiratory virus-associated airflow decline seems to be specific to viral species and infection localization. PMID- 16703506 TI - Human metapneumovirus infection induces long-term pulmonary inflammation associated with airway obstruction and hyperresponsiveness in mice. AB - BACKGROUND: Human metapneumovirus (hMPV) is a newly described paramyxovirus that is associated with bronchiolitis, pneumonia, and asthma exacerbation. The objective of the present work was to study the duration of pulmonary inflammation and the functional consequences of infection with hMPV by use of a BALB/c mouse model. METHODS: BALB/c mice were inoculated with 1 x 10(8) TCID(50) of hMPV type A (C-85473), and viral persistence in lungs was assessed by reverse-transcription polymerase chain reaction for 154 days after infection. Pulmonary inflammation was characterized in histopathological experiments by use of a validated scoring system, and periodic acid-Schiff (PAS) staining of lung sections was used to document increased mucus production, also until day 154. Finally, respiratory functions were analyzed by taking plethysmographic measurements until day 70. RESULTS: Persistence of viral RNA and significant pulmonary inflammation were noted until day 154, whereas the findings for PAS staining suggested that mucus production was increased only until day 12. Maximal breathing difficulties occurred on day 5, and airway obstruction and hyperresponsiveness were still significant until at least day 70. CONCLUSION: Acute hMPV infection in BALB/c mice is associated with long-term pulmonary inflammation that leads to significant obstructive disease of the airways. This animal model will be of a great benefit in the evaluation of novel therapeutic and prophylactic modalities. PMID- 16703507 TI - Swine as a principal reservoir of hepatitis E virus that infects humans in eastern China. AB - BACKGROUND AND METHODS: Genotype IV hepatitis E virus (HEV) has been isolated from humans and swine. To study the relationship between the human and swine reservoirs, we estimated their respective viral burden, analyzed the genetic makeup of the virus populations, and assessed the risk of infection associated with swine farming. RESULTS: In 2 swine-farming districts of eastern China, 9.6% of swine and 0.3% of healthy human subjects excreted HEV in stool, as did 68.8% of patients with confirmed HEV infection. The virus population circulating in humans consisted of genotype I and at least 4 phylogenetically distinct subgroups of genotype IV viruses, 2 of which concurrently circulated among swine. Persons engaged in occupations related to swine farming were found to have a 74% higher risk of infection than those engaged in other occupations, and persons living in communities downstream of the swine farms were found to have a 29% higher risk of infection than persons living in communities upstream. CONCLUSIONS: Genotype IV HEV is freely transmitted between humans and swine. Because the size of the swine population and its viral burden are much larger than those of humans, transmission of the virus most likely is directed from swine to humans. Infection can be acquired through contact with swine and their waste. PMID- 16703505 TI - Genetic diversity of human metapneumovirus over 4 consecutive years in Australia. AB - The molecular epidemiologic profile of human metapneumovirus (hMPV) infection has likely been skewed toward certain genetic subtypes because of assay-design issues, and no comprehensive studies have been conducted to date. Here, reverse transcription polymerase chain reaction was used to screen 10,319 specimens from patients presenting to hospitals with suspected respiratory tract infections during 2001-2004. After analysis of 727 Australian hMPV strains, 640 were assigned to 1 of 4 previously described subtypes. hMPV was the most common pathogen detected, and subtype B1 was the most common lineage. Concurrent, annual circulation of all 4 hMPV subtypes in our study population was common, with a single, usually different hMPV subtype predominating in each year. PMID- 16703508 TI - Postexposure protection of guinea pigs against a lethal ebola virus challenge is conferred by RNA interference. AB - BACKGROUND: Ebola virus (EBOV) infection causes a frequently fatal hemorrhagic fever (HF) that is refractory to treatment with currently available antiviral therapeutics. RNA interference represents a powerful, naturally occurring biological strategy for the inhibition of gene expression and has demonstrated utility in the inhibition of viral replication. Here, we describe the development of a potential therapy for EBOV infection that is based on small interfering RNAs (siRNAs). METHODS: Four siRNAs targeting the polymerase (L) gene of the Zaire species of EBOV (ZEBOV) were either complexed with polyethylenimine (PEI) or formulated in stable nucleic acid-lipid particles (SNALPs). Guinea pigs were treated with these siRNAs either before or after lethal ZEBOV challenge. RESULTS: Treatment of guinea pigs with a pool of the L gene-specific siRNAs delivered by PEI polyplexes reduced plasma viremia levels and partially protected the animals from death when administered shortly before the ZEBOV challenge. Evaluation of the same pool of siRNAs delivered using SNALPs proved that this system was more efficacious, as it completely protected guinea pigs against viremia and death when administered shortly after the ZEBOV challenge. Additional experiments showed that 1 of the 4 siRNAs alone could completely protect guinea pigs from a lethal ZEBOV challenge. CONCLUSIONS: Further development of this technology has the potential to yield effective treatments for EBOV HF as well as for diseases caused by other agents that are considered to be biological threats. PMID- 16703509 TI - Protection of Rhesus monkeys against dengue virus challenge after tetravalent live attenuated dengue virus vaccination. AB - Rhesus monkeys develop viremia after dengue virus (DENV) inoculation and have been used as an animal model to study DENV infection and DENV vaccine candidates. We evaluated, in this model, the protective efficacy of a live attenuated tetravalent DENV vaccine (TDV) candidate against parenteral challenge with parental near-wild-type DENV strains. Twenty monkeys were vaccinated with TDV at 0 and 1 month, and 20 unvaccinated monkeys served as controls. Vaccinated animals and their controls were inoculated with 10(3)-10(4) pfu of challenge virus 4.5 months after the second vaccination. Primary vaccination resulted in 95%, 100%, 70%, and 15% seroconversion to DENV serotypes 1, 2, 3, and 4 (DENV-1, -2, -3, and -4), respectively. After the second vaccination, the seropositivity rates were 100%, 100%, 90%, and 70%, respectively. Vaccination with TDV resulted in complete protection against viremia from DENV-2 challenge and in 80%, 80%, and 50% protection against challenge with DENV-1, -3, and -4, respectively. Our results suggest that the TDV can elicit protective immunity against all 4 DENV serotypes. Interference among the 4 vaccine viruses may have resulted in decreased antibody responses to DENV-3 and -4, which would require reformulation or dose optimization to minimize this interference during testing of the vaccine in humans. PMID- 16703510 TI - Immunization with a tetraepitopic lipid core peptide vaccine construct induces broadly protective immune responses against group A streptococcus. AB - BACKGROUND: The development of a vaccine to prevent infection with group A streptococcus (GAS) is hampered by the widespread diversity of circulating GAS strains and M protein types, and it is widely believed that a multivalent vaccine would provide better protective immunity. METHODS: We investigated the efficacy of incorporating 3 M protein serotypic amino-terminal epitopes from GAS isolates that are common in Australian Aboriginal communities and a conformational epitope from the conserved carboxy-terminal C-repeat region into a single synthetic lipid core peptide (LCP) vaccine construct in inducing broadly protective immune responses against GAS after parenteral delivery to mice. RESULTS: Immunization with the tetraepitopic LCP vaccine construct led to high titers of systemic, antigen-specific IgG responses and the induction of broadly protective immune responses, as was demonstrated by the ability of immune serum to opsonize multiple GAS strains. Systemic challenge of mice with a lethal dose of GAS given 60 or 300 days after primary immunization showed that, compared with the control mice, the vaccinated mice were significantly protected against GAS infection, demonstrating that the vaccination stimulated long-lasting protective immunity. CONCLUSIONS: These data support the efficacy of the LCP vaccine delivery system in the development of a synthetic, multiepitopic vaccine for the prevention of GAS infection. PMID- 16703511 TI - Use of DNA arrays to identify a mutation in the negative regulator, csrR, responsible for the high virulence of a naturally occurring type M3 group A streptococcus clinical isolate. AB - We previously reported that type M3 group A streptococcus (GAS) showed a wide range of 50% lethal dose values in mice. Analysis using DNA arrays indicated that the most virulent strain, M3-f, expressed significantly higher levels of the products of several virulence genes than did the other M3 isolates. Sequencing of the csrS, csrR, luxS, and rgg genes in the isolates showed that the M-3f csrR gene contained a specific point mutation. Disruption of wild-type (wt) csrR in an M3 strain increased its virulence and the expression of hyaluronic acid, whereas complementation with wt but not type M3-f csrR attenuated these changes. Expression experiments showed that type M3-f CsrR counteracted the effects of wt CsrR. Although wt CsrR bound to the hasA promoter region, type M3-f CsrR did not. Thus, the high virulence of the type M3-f strain is associated with the decreased binding of type M3-f CsrR to its target sequences. PMID- 16703512 TI - Group A streptococcal myonecrosis: increased vimentin expression after skeletal muscle injury mediates the binding of Streptococcus pyogenes. AB - Necrotizing fasciitis and myonecrosis caused by invasive infection with group A streptococci (GAS) are life-threatening conditions that have reemerged worldwide. Half of all GAS myonecrosis cases have no known portal of entry; yet, for unknown reasons, infection becomes established precisely at the site of a prior, nonpenetrating minor injury, such as a muscle strain. We hypothesized that GAS establishes infection by binding to surface molecules that are up-regulated on injured skeletal-muscle cells. Here, we isolated and identified vimentin as the major skeletal-muscle GAS-binding protein. Furthermore, we found that vimentin expression was up-regulated on injured skeletal-muscle cells in vitro and was expressed in muscle tissues from a patient with GAS myonecrosis who died of streptococcal toxic shock syndrome. These findings provide a molecular mechanism to explain the development of severe GAS soft-tissue infections at the sites of prior minor muscle trauma. This understanding may provide a basis for novel preventive strategies or therapies for patients with this devastating infection. PMID- 16703513 TI - Myeloid differentiation primary response gene (88)- and toll-like receptor 2 deficient mice are susceptible to infection with aerosolized Legionella pneumophila. AB - BACKGROUND: Toll-like receptors (TLRs) are a family of proteins that orchestrate innate immune responses to microbes. Although pathogens are typically recognized by multiple TLRs, the specific roles of individual TLRs in mediating host protection during in vivo infection are not well understood. METHODS: We compared the roles of myeloid differentiation primary response gene (88) (MyD88), which regulates signaling through multiple TLRs, and TLR2 in mediating resistance to aerosolized Legionella pneumophila infection in vivo. RESULTS: In comparison with wild-type mice, MyD88-deficient (MyD88(-/-)) mice had dramatically higher bacterial counts in the lungs, with decreased neutrophil counts in the bronchoalveolar lavage fluid as well as absent cytokine and chemokine production at early time points. By day 6 of infection, the MyD88(-/-) mice developed organizing pneumonia with dissemination of L. pneumophila to the lymph nodes and spleen. TLR2(-/-) mice were also more susceptible to L. pneumophila, with higher bacterial counts in the lung. However, TLR2(-/-) mice produced proinflammatory cytokines, recruited neutrophils to the lung alveoli, and cleared the infection without progression to organizing pneumonia and disseminated disease. CONCLUSIONS: These results suggest that a MyD88-dependent pathway is required for eradication of L. pneumophila and prevention of organizing pneumonia. TLR2 mediates partial resistance to L. pneumophila, which indicates that additional MyD88-dependent, TLR2-independent pathways are essential for full protection. PMID- 16703514 TI - A multidrug-resistant, acr1-deficient clinical isolate of Mycobacterium tuberculosis is unimpaired for replication in macrophages. AB - Multidrug-resistant tuberculosis (MDR-TB) poses a serious threat to global public health. The mutations responsible for drug resistance in Mycobacterium tuberculosis have been identified, but what impact these mutations have on bacterial fitness is controversial. We analyzed 3 MDR strains of M. tuberculosis obtained from human immunodeficiency virus-negative patients with chronic pulmonary TB. One of these strains harbored a chromosomal deletion encompassing 15 open reading frames. Genes deleted in this strain included acr1, which encodes the virulence factor alpha-crystallin (Acr) 1, a protein that has been reported to be essential for M. tuberculosis replication in macrophages. We found that all 3 MDR isolates, including the acr1-deficient strain, replicated in cultured murine and human macrophages with the same kinetics as H37Rv, a virulent laboratory strain. These observations challenge the prevailing view that MDR bacteria are less fit than drug-susceptible bacteria and indicate that Acr1 is dispensable for bacterial growth in the human lung. PMID- 16703515 TI - Autoimmunohistochemistry: a new method for the histologic diagnosis of infective endocarditis. AB - BACKGROUND: Although the pathologic examination of cardiac valves remains the reference standard for the diagnosis of infective endocarditis, the detection of microorganisms often poses a challenge for pathologists. This can be done by use of nonspecific histochemical stains or by immunohistochemical analysis, but specific antibodies are often not available. We describe a novel method for the detection of microorganisms in valve specimens from patients with infective endocarditis. METHODS: Detection of microorganisms was performed in valve specimens from patients with endocarditis caused by gram-positive cocci (25 specimens), blood culture-negative endocarditis (15 specimens: 6 cases caused by Coxiella burnetii, 5 caused by Tropheryma whipplei, and 4 caused by Bartonella species), or noninfective degenerative damage (30 specimens, used as negative controls), using the patients' own serum. This technique, called "autoimmunohistochemistry," is an immunohistochemical peroxidase-based method that we compared with results of culture and polymerase chain reaction (PCR) assay. RESULTS: Bacteria were detected by autoimmunohistochemistry in 20 (80%) specimens from patients with endocarditis caused by gram-positive cocci and in 15 (100%) specimens from patients with blood culture-negative endocarditis but in no control specimens. The rate of detection of bacteria by autoimmunohistochemistry was significantly higher than that by culture but was similar to that by PCR. CONCLUSIONS: Autoimmunohistochemistry may be useful for the detection of microorganisms in samples of valves from patients with infective endocarditis. This new diagnostic tool may be particularly useful in cases of blood culture negative endocarditis. PMID- 16703516 TI - Long-term decay of the HIV-1 reservoir in HIV-1-infected children treated with highly active antiretroviral therapy. AB - To investigate the decay of the human immunodeficiency virus type 1 (HIV-1) reservoir in children receiving highly active antiretroviral therapy (HAART), we measured HIV-1 DNA in peripheral blood mononuclear cells from 14 children who achieved and maintained suppression of plasma viremia up to 48 months after the initiation of HAART. Levels of intracellular unspliced and multiply spliced HIV-1 RNA were used as markers of residual viral replication. During the first month of HAART, there were significant decays in levels of both plasma HIV-1 RNA and multiply spliced HIV-1 RNA, yet unspliced HIV-1 RNA persisted in most of the children. Greater HIV-1 DNA decay during the first month of HAART correlated with a higher concomitant increase in CD4(+) cell counts (P=.028) and a smaller subsequent HIV-1 DNA decay (P=.0012). Furthermore, HIV-1 DNA decayed faster from 1 to 9 months of HAART (median half-life, 5 months) than during the subsequent follow-up period (median half-life, 30 months). Moreover, after 9 months of HAART, HIV-1 DNA tended to decay more slowly in children with detectable levels of unspliced HIV-1 RNA. These findings suggest that clearance of the viral reservoir in HAART-treated children may be influenced by immune repopulation and residual viral replication and may help in refining long-term treatment strategies. PMID- 16703517 TI - Detection of ganciclovir resistance in patients with AIDS and cytomegalovirus retinitis: correlation of genotypic methods with viral phenotype and clinical outcome. AB - BACKGROUND: The cytomegalovirus (CMV) UL97 gene can be sequenced either from blood specimens directly amplified by polymerase chain reaction (PCR) or from culture isolates, to detect resistance to ganciclovir. METHODS: A prospective epidemiological study was conducted in which paired specimens were routinely obtained for sequencing of the UL97 gene from blood specimens (i.e., plasma and leukocytes) directly amplified by PCR and from CMV culture isolates. The specimens then were compared with each other and in terms of results of susceptibility testing and their association with progression of retinitis. RESULTS: A total of 845 paired specimens were obtained from 165 patients with AIDS and CMV retinitis. There typically was >90% agreement between the UL97 gene sequences from blood specimens directly amplified by PCR and those from culture isolates. The agreement between phenotypic resistance and the detection of UL97 mutations was >92% for PCR-amplified blood specimens and >97% for culture isolates. Plasma and leukocytes performed similarly. Progression of retinitis was correlated with the detection of UL97 mutations in PCR-amplified blood specimens, with adjusted odds ratios of 7.02 (P=.002) for leukocytes, 9.11 (P=.02) for plasma, and 17.6 for culture isolates (P<.0001). CONCLUSIONS: Because blood specimens directly amplified by PCR can be analyzed more rapidly than can cultures (< or =48 h vs. > or =4 weeks), sequencing the CMV UL97 gene from blood specimens directly amplified by PCR may be useful clinically. PMID- 16703518 TI - Occurrence of the Southeast Asian/South American SVMNT haplotype of the chloroquine-resistance transporter gene in Plasmodium falciparum in Tanzania. AB - Two main haplotypes, CVIET and SVMNT, of the Plasmodium falciparum chloroquine resistance transporter gene (Pfcrt) are linked to 4-aminoquinoline resistance. The CVIET haplotype has been reported in most malaria-endemic regions, whereas the SVMNT haplotype has only been found outside Africa. We investigated Pfcrt haplotype frequencies in Korogwe District, Tanzania, in 2003 and 2004. The SVMNT haplotype was not detected in 2003 but was found in 19% of infected individuals in 2004. Amodiaquine use has increased in the region. The introduction and high prevalence of the SVMNT haplotype may reflect this and may raise concern regarding the use of amodiaquine in artemisinin-based combination therapies in Africa. PMID- 16703520 TI - Investigating new antiretroviral combinations. PMID- 16703522 TI - No role for thalidomide in the treatment of leprosy. PMID- 16703524 TI - [Language and cognitive impairment: a semiological study into visual naming]. AB - INTRODUCTION: The presence of anomia and/or paraphasias in patients with cognitive impairment suggests an associated deterioration of their language. Visual naming makes it possible to examine these signs in a controlled manner as the target word is already known. AIM. To conduct a semiological study of naming in normal aging, cognitive impairment and language impairment conditions. SUBJECTS AND METHODS: The study consisted in an analysis of nine types of signs (correct response, increased latencies, circumlocutions, absence of response, semantic verbal paraphasias, verbal paraphasias with a similar form, unrelated verbal paraphasias, phonemic paraphasias and neologisms) in a visual naming task (6 items), in two groups with normal language--controls > 70 years and patients with Alzheimer's disease (AD) with onset of amnesia--and two groups with language impairment--vascular aphasia and anomic AD-. RESULTS: Patients failed to perform naming correctly in 4.2% of the responses in controls, 10% in patients with amnesic AD, 30% in patients with vascular aphasia and 50% in patients with anomic AD. Semantic paraphasias were observed in the two groups with normal language abilities (controls and amnesic AD), although frequencies were low. Signs that suggest difficulties in accessing/retrieving lexical items were more frequent in patients with AD and ran parallel to the degree of anomia. One notable finding was the absence of signs of phonological dysfunction in the two groups with AD, regardless of the degree of anomia. CONCLUSIONS: Semiological quantification makes it possible to distinguish differences in the degree of anomia and in the pattern of errors both in controls and in patients with amnesic onset AD and among patients with vascular aphasia and neurodegenerative anomia. PMID- 16703525 TI - [Hospital care for stroke patients in the Valencian Region using the basic minimum data set from the International Classification of Diseases, 9th revision, clinical modification]. AB - AIM: To determine the load and characteristics of cerebrovascular diseases (CVD) admitted in the hospital network throughout the Valencian Region. PATIENTS AND METHODS: The paper reports on an analysis of the information included in the basic minimum data set (BMDS) from the 26 hospitals run by the Valencian Regional Ministry of Health in the year 2001. Patients that were selected were those whose main diagnosis was codes C.430 to C.437, according to the International Classification of Diseases, 9th revision, clinical modification (ICD-9-CM). RESULTS: A total of 10,558 patients with CVD were discharged, which accounts for 2.6% of admissions and 3% of hospital stays. The mean age of the series was 71.03 years -standard deviation (SD): 9- and 94.8% were admitted as emergencies. By diagnoses, 3% (319) were subarachnoid haemorrhages (SAH; C.430); 13.4% (1,412) were cerebral haemorrhages (ICH; C.431); 18.5% (1,956) were transient ischemic attacks (TIA; C.435); 49.5% (5,225) were cases of cerebral infarction (CI; C.434 and C.436); and 15.6% involved other vascular processes (C.433 and C.437). Mortality rates were 30.1% in SAH; 33.9% in ICH; 11.7% in CI; and 2.7% in TIA. Mean number of days in hospital: SAH 17.4 (SD: 15); ICH 13.1 (SD: 11.8); CI 9.9 (SD: 6.4) and in cases of TIA 7.2 (SD: 4). The percentages of survivors who were discharged home were 78.9 % in SAH, 83.2% in ICH and 91.9% in the case of CI. In all, 51.3% (5,413 patients) were discharged by neurological units. CONCLUSION: In spite of possible insufficiencies analysed in this work, the use of the BMDS provides valuable epidemiological information that is very useful for health care management. PMID- 16703526 TI - [The functional condition of patients following a cerebrovascular accident]. AB - INTRODUCTION: Most patients who suffer a cerebrovascular accident (CVA) belong to the adult-elderly age group. Thus, such patients are likely to display a certain degree of functional compromise as a result of the natural process of aging, regardless of whether they have suffered a CVA or not. AIM. To examine the functional condition of patients with CVA, as well as that of healthy individuals who have never suffered a stroke. SUBJECTS AND METHODS: An analytical cross sectional study was conducted with a control group made up of 82 individuals -41 of whom were CVA patients (G1) and 41 were individuals with no history of CVA (G2)-. These subjects were paired according to sex, age and educational level. RESULTS: The sample was predominantly made up of females (60%), who were married (70%), with ischaemic CVA (70%), in the right hemisphere of the brain (56%) and with a primary education (36%). The prevalence rate of patients with severe functional impairment was 58.53%, whereas in the control group there were no subjects with this level of deficiency. A relationship was found to exist (p < 0.05) between the functional condition and the variables age and causation of CVA for G1, and also between the functional condition and the variable age for the individuals in G2, which indicates that functional compromise is greater in more elderly subjects. CONCLUSIONS: Findings suggest that the functional compromise of patients with CVA is higher than that of individuals with no history of CVA with the same sex, age and educational level. This clearly shows that there is an association between CVA and functional impairment in these patients. PMID- 16703527 TI - [Unusual cortical compromise in a case of Wernicke's encephalopathy]. AB - INTRODUCTION: Wernicke's encephalopathy (WE) is a metabolic disease due to thiamine deficiency; only 10% of cases are diagnosed pre-mortem. Symptoms of WE include ophthalmoplegia, nistagmus, ataxia and mental confusion; post-mortem examination shows characteristic symmetrical lesions in the mamillary bodies (MB), hypothalamus, thalamus, brain stem and cerebellum with spongiosis, demyelination, vascular proliferation and relative preservation of neurons. CASE REPORT: 50 years-old male with alcoholic hepatopathy and orthotopic hepatic transplant who suffered a second surgical intervention 10 days after due to problems in the biliar anastomosis. After this second surgery he showed an altered mental status, with fluctuating global confusion, disorientation and agitation. He died 52 days after the hepatic transplantation. Autopsy study showed bilateral broncopneumonia, brown discoloration of the MB and bilateral linear lesions in the cortex of both motor gyri, which histologically showed identical to the MB lesions with demyelination, capillary and glial proliferation and preservation of neurons. Alzheimer type II astrocytes were also found in basal nuclei and cortex. CONCLUSIONS: Typical WE lesions affect MB, hypothalamus, thalamus, brain stem and cerebellum; cortical lesions, when found, are due to hepatocerebral degeneration with Alzheimer type II astrocytes or to the citopathic effects of ethanol. In our case, cortical lesions were identical to the lesions found in MB, an extraordinary finding which we have not found reported in the literature. PMID- 16703528 TI - [Attention deficit hyperactivity disorder in adults: a clinical and therapeutic characterization]. AB - AIM: To update the existing knowledge about attention deficit hyperactivity disorder (ADHD) in adults, with special interest given to aspects concerning epidemiology, diagnosis, progression and treatment. DEVELOPMENT: Acknowledging the fact that ADHD can persist into adulthood is a relatively recent development. Nevertheless, over the last few years evidence has been put forward to support the validity of its diagnosis in adults. The prevalence of ADHD in the general adult population is estimated to be around 4%. Over 50% of the children with the disorder will continue to have it as adults. The symptoms of ADHD can be different in adults, with less hyperactivity and a slight decrease in impulsiveness, but with the same symptoms of inattention. ADHD in adulthood is linked to serious economic, occupational, academic and familial repercussions, as well as to road accidents and to the presence of other psychiatric pathologies (such as drug addiction, personality disorders and depression). CONCLUSIONS: A number of different psychometric instruments are available in Spanish for evaluating ADHD in adults. The use of structured interviews, together with a systemized evaluation of possible comorbid disorders, makes it easier to reach a correct differential diagnostic. Pharmacological treatment in adults is essentially the same as that used with children, but weight-adjusted. Clinical trials have been carried out with psychostimulants and results show methylphenidate to be the most effective and the safest. Atomoxetine is the non stimulating pharmaceutical that has been most widely studied in adults. Cognitive behavioural psychological treatments have proved to be effective in adults with ADHD. PMID- 16703529 TI - [Glutamate-related excitotoxicity neuroprotection with memantine, an uncompetitive antagonist of NMDA-glutamate receptor, in Alzheimer's disease and vascular dementia]. AB - AIM: To review the therapeutic efficacy of memantine, an uncompetitive antagonist of N-methyl-D-aspartate (NMDA)-glutamate receptor. DEVELOPMENT: Alzheimer's disease (AD) is the most common neurodegenerative disorder and cause of dementia with ageing worldwide. The main AD symptoms are a gradual loss of cognitive function and a functional impairment. Glutamatergic excitatory neurotransmission, an important process in learning and memory, is severely disrupted in AD, probably due to the oxidative stress associated with the beta-amyloid peptide (1 42) increase. The glutamate-related excitotoxicity, mainly mediated by NMDA subtype of the glutamate receptors, is a common clue of pathogenesis for neurodegenerative disorders. CONCLUSIONS: Memantine, a moderate-affinity, voltage dependent, uncompetitive antagonist of NMDA receptor, shows neuroprotective effects in patients with moderate-to-severe AD. Memantine is a drug with neuroprotective and cognition-enhanced properties, which can be combined with other treatments for AD. Thus, memantine does not stop or reverse AD, but its moderating effect in protecting the brain from the toxic levels of calcium, allows normal signaling among brain neurons. The efficacy and safety profile of memantine have been reported in several clinical trials for treatment of AD and vascular dementia. PMID- 16703530 TI - [Guillain-Barre syndrome as a complication of epidural anaesthesia]. PMID- 16703531 TI - [The progression of familial amyloidotic polyneuropathy following a liver transplant]. PMID- 16703532 TI - [Medullary infarct due to methamphetamine]. PMID- 16703533 TI - [Marchiafava-Bignami disease in a non-alcoholic patient]. PMID- 16703534 TI - Mikulicz's disease: a new perspective and literature review. AB - PURPOSE: To report the clinical and pathophysiologic features of two patients with Mikulicz's disease and to further characterize recommendations for diagnosis and management with a review of the literature. METHODS: Retrospective nonrandomized consecutive case series, Jules Stein Eye Institute, David Geffen School of Medicine at UCLA. RESULTS: Mikulicz's disease is characterized by symmetric lacrimal, parotid, and submandibular gland enlargement with associated lymphocytic infiltrations. The authors noted two cases of Mikulicz's disease. The diagnosis of Mikulicz's disease was based on the following criteria: 1) symmetric and persistent swelling of the lacrimal glands and either or both of the major salivary glands (parotid and submandibular); and 2) the exclusion of other diseases that may mimic this presentation, such as sarcoidosis, viral infection, or lymphoproliferative disorders. CONCLUSIONS: Mikulicz's disease is a condition in which there is bilateral lacrimal and salivary gland swelling that is not associated with other systemic conditions. The condition is self-limiting and most often, the diagnosis is a clinical one. Previously, Mikulicz's disease was often considered as a subtype of Sjogren's syndrome (SS). Clinical and immunologic differences between Mikulicz's disease and SS may warrant further consideration of Mikulicz's disease as a specific autoimmune phenomenon separate from SS, and Mikulicz's disease may be amenable to different treatment modalities than those employed in patients with SS. PMID- 16703535 TI - Apraxia of eyelid opening: clinical features and therapy. AB - PURPOSE: Botulinum toxin injection is the treatment of choice in cases of benign essential blepharospasm. However, about 10% of the patients do not get sufficient effect from this treatment, and many of them have concomitant apraxia of lid opening. METHODS: Over a 3-year period we treated 12 patients. Three had pure apraxia of lid opening and in the other nine it was associated with blepharospasm. All patients were initially treated with botulinum toxin injections with poor results. They underwent surgical treatment like blepharoplasty, limited myectomy, aponeurosis repair, and/or frontalis suspension. Some of them needed post operative botulinum toxin injections in the pretarsal part of orbicularis oculi muscles. RESULTS: This combined therapy gave good functional and aesthetic results. CONCLUSIONS: The specific causes of blepharospasm and apraxia of lid opening are unknown, but these two conditions coexist in some patients and can be difficult to treat. It is important to make a correct diagnosis, and a combined surgical and botulinum toxin treatment can be very effective. PMID- 16703536 TI - Limbal conjunctival mini-autografting for preventing recurrence after pterygium surgery. AB - PURPOSE: The aim of the present study was to evaluate limbal conjunctival mini autografting procedure based on recurrence rates following pterygium surgery. METHODS: A total of 63 eyes (53 patients) with pterygium, of which 60 were primary and 3 were recurrent, underwent surgery for the removal of pterygia with limbal conjunctival mini-autograft. After the pterygium excision, the limbal portion of the graft was oriented and sutured to the limbus at the recipient bed with the epithelial surface upside. Recurrence was defined as fibrovascular tissue of more than 1 mm over the cornea in the area of previous pterygium excision. The mean follow-up period was 12.5 months (ranging between 6 and 29 months). RESULTS: The time required for the surgery was approximately 30 minutes (range 20-40 minutes). Recurrence of pterygium was observed in 6 of 63 eyes (9.52%). No eyes with recurrent pterygium developed recurrence postoperatively. There were no severe complications during the operative and postoperative period. Most patients had temporary graft edema, lasting a few days after the operation, and they experienced and complained of foreign body sensations and epiphora. CONCLUSIONS: Limbal conjunctival mini-autografting appears to be an effective procedure for pterygium surgery resulting in a low recurrence rate. PMID- 16703537 TI - Visual outcome after excimer laser refractive surgery in adult patients with amblyopia. AB - PURPOSE: To evaluate the visual and refractive outcome in adult patients wi th refractive and anisometropic amblyopia treated with photorefractive keratectomy. METHODS: An interventional non-comparative study was performed on 50 consecutive patients with amblyopia treated with photorefractive keratectomy. Sixty-eight eyes of 18 participants with bilateral refractive amblyopia and 32 participants with unilateral anisometropic amblyopia were studied. The photorefractive keratectomy was performed with MEL 70 G-Scan excimer laser. Follow-up was 1, 3, 6, and 12 months after the treatment. The mean outcome measures were uncorrected visual acuity (UCVA), best spectacle corrected visual acuity (BCVA), refraction, and corneal transparency. RESULTS: In 64.7% of eyes, the UCVA at 12 months after the treatment was better than or equal to their preoperative BCVA. A total of 82.4% of eyes improved their BCVA one or more lines. No eye lost lines of BCVA, 17.6% remained unvaried, 50% gained one line, 20.6 % gained two lines, and 11.8% gained three lines of BCVA. The number of lines gained correlated strongly with diopters of anisometropia between eyes (p<0.01). The mean sphere changed from 5.62+/-3.37 D to -0.47+/-1 D, mean cylinder varied from -2.36+/-1.57 to -0.18+/ 0.41 D, and the spherical equivalent varied from -6.82+/-3.1 to -0.45+/-1.08 D. No clinically significant haze was observed. CONCLUSIONS: Excimer laser refractive surgery appears to be a safe and efficient procedure in the treatment of ametropic and anisometropic amblyopia in adults. Adult patients with anisometropic and refractive amblyopia could benefit from the refractive treatment with significant improvement of the visual acuity. PMID- 16703538 TI - Zyoptix wavefront-guided versus standard photorefractive keratectomy (PRK) in low and moderate myopia: randomized controlled 6-month study. AB - PURPOSE: To evaluate the refractive and aberrometric outcome of wavefront-guided photorefractive keratectomy (PRK) compared to standard PRK in myopic patients. METHODS: Fifty-six eyes of 56 patients were included in the study and were randomly divided into two groups. The study group consisted of 28 eyes with a mean spherical equivalent (SE) of -2.25+/-0.76 diopters (D) (range: -1.5 to -3.5 D) treated with wavefront-guided PRK using the Zywave ablation profile and the Bausch & Lomb Technolas 217z excimer laser (Zyoptix system) and the control group included 28 eyes with a SE of -2.35+/-1.01 D (range: -1.5 to -3.5 D) treated with standard PRK (PlanoScan ablation) using the same laser. A Zywave aberrometer was used to analyze and calculate the root-mean-square (RMS) of total high order aberrations (HOA) and Zernike coefficients of third and fourth order before and after (over a 6-month follow-up period) surgery in both groups. Preoperative and postoperative SE, un-corrected visual acuity (UCVA), and best-corrected visual acuity (BCVA) were evaluated in all cases. RESULTS: There was a high correlation between achieved and intended correction. The differences between the two treatment groups were not statistically significant for UCVA, BCVA, or SE cycloplegic refraction . Postoperatively the RMS value of high order aberrations was raised in both groups. At 6-month control, on average it increased by a factor of 1.17 in the Zyoptix PRK group and 1.54 in the PlanoScan PRK group (p=0.22). In the Zyoptix group there was a decrease of coma aberration, while in the PlanoScan group this third order aberration increased. The difference between postoperative and preoperative values between the two groups was statistically significant for coma aberration (p=0.013). No statistically significant difference was observed for spherical-like aberration between the two groups. In the study group eyes with a low amount of preoperative aberrations (HOA RMS lower than the median value; <0.28 microm) showed an increase of HOA RMS while eyes with RMS higher than 0.28 microm showed a decrease (p<0.05). CONCLUSIONS: Zyoptix wavefront-guided PRK is as safe and efficacious for the correction of myopia and myopic astigmatism as PlanoScan PRK. Moreover this technique induces a smaller increase of third order coma aberration compared to standard PRK. The use of Zyoptix wavefront-guided PRK is particularly indicated in eyes with higher preoperative RMS values. PMID- 16703539 TI - Photorefractive keratectomy in high myopic defects with or without intraoperative mitomycin C: 1-year results. AB - PURPOSE: To study the results of the prophylactic use of mitomycin C (MMC) to reduce haze formation and refractive regression after excimer laser photorefractive keratectomy (PRK) for high myopic defects (>5 diopters). METHODS: Prospective, consecutive, observational study. A total of 124 eyes of 62 patients were divided into two groups of 31 patients, 62 eyes each (Groups A and B). Only Group A was treated with MMC 0.02%. The data of the two groups of eyes, related to the best-corrected visual acuity (BCVA), to the difference of refraction pre- and post-treatment, and to the corneal haze, were analyzed through combined permutation tests by using the NPC Test software . RESULTS: BCVA of Group A, 1 year after treatment, was better than that of the control Group B (one-sided p value = 0.013): Group A - 3 eyes (4.8%) had a loss of a decimal fraction and no eyes > 1; Group B - 13 eyes (20.9%) had a loss of a decimal fraction and 1 eye (1.6%) of 2. There was a smaller difference between attempted and achieved SE correction in Group A with respect to Group B (one-sided p value = 0.068): Group A - 43 eyes (69.3%) within +/- 0.50 D; Group B - 31 eyes (50%) within +/- 0.50 D. there was a smaller incidence of corneal haze in the group for which MMC was used (one-sided p value = 0.005). CONCLUSIONS: In this study, the application of MMC 0.02% solution immediately after PRK produced lower haze rates and had better predictability and improved efficacy 1 year after treatment. PMID- 16703540 TI - Quantitative corneal anatomy in emmetropic subjects. AB - PURPOSE: Currently there is little information available about the corneal thickness values of healthy emmetropic subjects. Therefore, the authors decided to analyze the corneal thickness in healthy emmetropic subjects. METHODS: The authors analyzed the difference in thickness values between the thinnest corneal site and the central and paracentral cornea in 124 eyes of 124 healthy emmetropic white subjects. RESULTS: The mean difference between the thinnest site of the cornea and the thickness values obtained in the areas analyzed was as follows: 12+/-6 microm center; 140+/-19 microm superonasal; 133+/-23 microm nasal; 117+/ 26 microm inferonasal; 122+/-19 microm superotemporal ; 89+/-22 microm temporal; and 99+/-29 microm inferotemporal (p<0.001; one way analysis of variance test). CONCLUSIONS: In healthy emmetropic white subjects the thinnest site of the cornea is statistically lower than the central and paracentral cornea. PMID- 16703541 TI - Comparison of the effect of sodium hyaluronate (Ophthalin) and hydroxypropylmethylcellulose (HPMC-Ophtal) on corneal endothelium, central corneal thickness, and intraocular pressure after phacoemulsification. AB - PURPOSE: To prospectively evaluate the effects of 2% hydroxypropyl methylcellulose (HPMC-Ophtal) and sodium hyaluronate 1% (Ophthalin) on intraocular pressure, corneal thickness, and endothelial cell loss in small incision cataract surgery with implant. METHODS: A total of 110 patients undergoing routine phacoemulsification with implant received either 2% hydroxypropyl methylcellulose or sodium hyaluronate 1% as ophthalmic viscosurgical device. Pre- and postoperative slitlamp examination, intraocular pressure measurement (preoperatively and at 1-4 hours, 1 day, and 7 days postoperatively), ultrasonic pachymetry (preoperatively and at 1 week, 4-6 weeks, and 12 weeks post operatively), and corneal endothelial cell count (preoperatively and 12 weeks postoperatively) were performed. Data were analyzed using two-way analysis of variance. RESULTS: All measurements were comparable between the two groups preoperatively. Intraocular pressure was significantly lower in the Ophthalin group at 1 day post operatively, while no significant difference was found between the two groups on the 1-4 hours and 7 days examination. The central corneal thickness was not significantly different between the two groups at any postoperative visit . However, the mean cell density demonstrated a significant fall of 11.76% for Ophthalin and 4.27% for HPMC-Ophtal at 12 weeks post-operatively, the difference between the two being significant (p=0.009). CONCLUSIONS: 2% Hydroxypropyl methylcellulose, compared with sodium hyaluronate 1%, is superior in protecting the corneal endothelial cells, has the same effect on central corneal thickness, and is associated with slightly higher intraocular pressure 1 day post operatively. It compares favorably with sodium hyaluronate 1% and can be used as an effective and cheaper alternative in routine small incision cataract surgery with implant. PMID- 16703542 TI - The comparison of eyelash lengthening effect of latanoprost therapy in adults and children. AB - PURPOSE: To compare the eyelash lengthening effect of latanoprost in adults and children with glaucoma. METHODS: Twenty eyes of 13 men and 7 women (mean age: 54.9, range 42-69 years) with primary open-angle glaucoma and 20 eyes of 9 boys and 11 girls (mean age: 10.7, range 6-16 years) with glaucoma were included in this prospective study. In 19 children, juvenile glaucoma and in one, pseudophakic glaucoma had been detected. A single eyelash was pulled from the center of the upper eyelid before latanoprost therapy and at the sixth month of therapy, and measured. RESULTS: In adult cases, the mean eyelash length was 5.79+/-0.18 mm (5.5-6.1 mm) at baseline and 6.45+/-0.21 mm (6.2-6.8 mm) at the sixth month. In children, the mean length was 5.66+/-0.22 mm (5.3-6.0 mm) at baseline and 6.39+/-0.37 mm (5.9-6.9 mm) at the sixth month. The mean difference in eyelash lengths at baseline and the sixth month was 0.67+/-0.09 mm (0.5-0.7 mm) in adults and 0.75+/-0.25 mm (0.4-1.2 mm) in children. CONCLUSIONS: The differences in mean eyelash lengths at baseline and at the sixth month of latanoprost therapy were statistically significant in both adults and children (p=0.000). The mean of the difference of the eyelash length in children was higher than in adults but the result was not statistically significant (p=0.678). PMID- 16703543 TI - Glaucoma detection using the GDx nerve fiber analyzer and the retinal thickness analyzer (RTA). AB - PURPOSE: To compare the ability of the nerve fiber analyzer (GDx) and the retinal thickness analyzer (RTA) to discriminate between glaucomatous and healthy eyes. METHODS: Thirty-seven glaucoma patients (early to moderate severity) and 34 healthy controls were included. Glaucoma patients were defined as those with two repeatable abnormal visual fields by automated perimetry within 1 year. All subjects were examined with a GDx scanning laser polarimeter and RTA. Twelve GDx retinal nerve fiber layer parameters and 12 RTA optic disk topography parameters were obtained. GDx and RTA measurements were compared between both experimental groups using t-tests. Areas under the receiver operating characteristic curves (AUROC) for discriminating between healthy and glaucomatous eyes using GDx and RTA parameters were calculated and compared, and sensitivities at >or=80% and >or=95% specificity were reported. RESULTS: Statistically significant differences between glaucomatous and healthy eyes were found for most GDx and RTA parameters. For GDx, the parameter with the largest AUROC for discriminating between healthy and glaucomatous eyes was the number (AUROC = 0.91, sensitivity = 85% at specificity = 84%, sensitivity = 73% at specificity = 95%). For RTA, the parameter with the largest AUROC was mean cup depth (AUROC = 0.79, sensitivity = 61% at specificity = 82%, sensitivity = 33% at specificity = 95%). The AUROC for the GDx number was significantly larger than the AUROC for RTA mean cup depth (p<0.05). CONCLUSIONS: GDx showed better discrimination and better sensitivities at fixed specificities than RTA. The currently available RTA optic disk analysis software likely cannot replace GDx RNFL analysis software for successful glaucoma diagnosis. PMID- 16703544 TI - Diagnostic accuracy and reproducibility of tendency oriented perimetry in glaucoma. AB - PURPOSE: To evaluate the diagnostic capability of tendency oriented perimetry (TOP) in glaucoma. METHODS: A): The diagnostic accuracy of mean defect (MD), square-root of the loss variance (s LV), and number of pathologic points (NPP) was calculated in 295 normal and 414 glaucoma eyes (179 early, 112 moderate, and 123 advanced) examined with TOP. B): Threshold fluctuation (F) and its relationship with the loss variance (LV) was measured in 34 normal and 33 glaucoma eyes (mean MD=3 dB; SD=3.9) for TOP and for full-threshold perimetry (FT). C): Twenty-eight eyes with stable glaucoma (mean MD=9.5 dB; SD=7.2) were examined six times to quantify LV error. D): TOP and FT were tested with the simulation program PeriSim using different behavior models. RESULTS: A): The best diagnostic index in early glaucoma (MD<6dB) was sLV (specificity=90.2%, sensitivity=84.9). The three indices had similar precision in moderate and severe glaucoma. B): Threshold fluctuation and sLV were better correlated in TOP (r=0.72, p<0.01) than in FT (r=0.62, p<0.01). For normal subjects, in FT the incidence of F<2 dB was 8.82% and s LV<1.5 dB 5.88%. The same frequencies in TOP were 67.65% and 55.88%. C): Averaging six examinations reduced the sLV val ue by 22%. D): The threshold estimation error increased 1 dB in TOP in relation to FT for the same patient's behavior, but the error in TOP was lower than i n FT when the worst behavior was modeled. CONCLUSIONS: TOP is a good discriminator between glaucoma and normality. Perimetry results overestimate the real sLV value. TOP's high diagnostic ability is probably associated to the algorithm design and to less contaminating influences during the examination. PMID- 16703545 TI - Pseudoexfoliation syndrome and its antioxidative protection deficiency as risk factors for age-related cataract. AB - PURPOSE: Pseudoexfoliation syndrome (PES) seems to be a systemic condition. However, to an ophthalmologist it represents a continual challenge, due to unclear etiology and patho-genesis, and because of a potential for development of an aggressive glaucoma, and cataract surgery complications. According to some findings, PES could be a factor of hastened aging, which means that it could be considered as a secondary aging factor. METHODS: Frequencies of secondary aging diseases (non-insulin dependent diabetes mellitus, atherosclerotic myocardiopathy, chronic obstructive lung diseases, arterial hypertension, and PES) were investigated in 162 patients with age-related cataract and 55 age- and sex-matched control subjects, and analyzed by a logistic regression. The authors also determined elements of antioxidative protection in a group of sera from patients with cataract and PES, and compared them to those obtained from patients with cataract without PES (activity of catalase and peroxidase, glutathione, vitamins C and E), and total antioxidative protection (%iMDA). Antioxidant defense of aqueous humors, expressed as a rate of induced malondialdehyde (%iMDA), and total thiol groups in the lens corticonuclear blocks (TSH) were estimated in samples of 17 patients with PES and cataract and 55 patients with cataract only. RESULTS: Logistic regression showed the highest odds ratio for PES (OR=4.516; p<0.05). Catalytic activity of serum catalase had significantly lower values in patients with PES (p<0.05). Anti oxidative defense of aqueous humor and lens had lower values in patients with PES. CONCLUSIONS: The results indicate that PES might be a significant factor for cataractogenesis. At least a part of pathogenesis alterations in an eye with PES could be the result of higher intensity of oxidative stress. PMID- 16703546 TI - Central serous chorioretinopathy and Helicobacter pylori. AB - PURPOSE: This study was designed to evaluate the prevalence of Helicobacter pylori infection in patients with central serous chorioretinopathy (CSC). METHODS: Retrospective observational case series. A group of 23 patients (22 men and 1 woman, age range 34-62 years, median age 47 years) with diagnosis of CSC, confirmed by fluorescein angiogram, and a control group of 23 consecutive patients (22 men and 1 woman, age range 41-69 years, median age 50 years) referred to our Department for retinal disease other than CSC were studied. Each patient provided peripheral venous blood samples and a stool specimen, which were analyzed at the Department of Gastroenterology and Microbiology at the same university. H. pylori infection was determined by measurement of IgG anti-bodies to H. pylori and by determination of H. pylori antigens in the stool specimens by enzyme-linked immunosorbent assay technique. Patients were defined as H. pylori infected if both tests were positive. RESULTS: The prevalence of H. pylori infection was 78.2% (95% CI, 56%-92%) in CSC patients and 43.5% (95% CI, 23%-65%) in control subjects (p<0.03 by two-tail ed Fisher exact test). The odds ratio for CSC associated with H. pylori infection was 4.6 (95% CI 1.28-16.9). CONCLUSIONS: The results of this study show that the prevalence of H. pylori infection seems to be significantly higher in patients with CSC than in control s. H. pylori infection may represent a risk factor in patients with CSC. PMID- 16703547 TI - Surgical results of combined pars plana vitrectomy, phacoemulsification, and intraocular lens implantation. AB - PURPOSE: To evaluate the results and complications of combined pars plana vitrectomy (PPV), phacoemulsification and aspiration (PEA), and intraocular lens (IOL) implantation. METHODS: A total of 117 eyes from 114 patients who had undergone PPV combined with PEA and IOL implantation were retrospectively analyzed. Combined surgery was performed for a wide variety of vitreoretinal diseases. Intraoperative and postoperative complications were also reviewed. RESULTS: The postoperative BCVA improved by 2 lines or more in 85 eyes (72.6%). Intraoperative complications consisted of retinal tears in 14 eyes (12.0%) and posterior capsular rupture in 2 eyes (1.7%). Iatrogenic retinal tears occurred more frequently in eyes with a macular hole than in eyes with any other disease (p=0.005, chi-square test). Postoperative complications consisted of posterior capsule opacification (PCO) (21 eyes), transient IOP elevation (29 eyes), vitreous hemorrhage (6 eyes), anterior chamber fibrin exudation (11 eyes), posterior iris synechia (8 eyes), neovascular glaucoma (1 eye), and recurrent retinal detachment (RD) (2 eyes). Fibrin exudation occurred more frequently in eyes with proliferative diabetic retinopathy (PDR) and RD than in eyes with any other disease (p=0.03, chi-square test). PCO occurred more frequently in eyes with PDR than in eyes with any other disease (p=0.03, chi-square test). CONCLUSIONS: The present study suggests that a high success rate can be achieved when recently improved PPV techniques are combined wi th PEA and IOL implantation. The complications that were observed following this combined treatment varied with respect to the vitreoretinal disease present prior to surgery. PMID- 16703548 TI - Preoperative factors influencing visual outcome following surgical excision of subfoveal choroidal. AB - PURPOSE: To evaluate long-term visual acuity outcomes and the influences of various preoperative factors on visual outcome in patients undergoing surgical removal of choroidal neovascular neovascularization (CNV) caused by age-related macular degeneration (ARMD). METHODS: The authors studied 146 eyes of 146 patients who were followed for at least 1 year after surgical excision of CNV associated with ARMD. Surgical indications included subfoveal active CNV localized mainly above the retinal pigment epithelium (RPE) and a standard Japanese decimal visual acuity of 0.3 or worse. CNV above the RPE was diagnosed by fluorescein angiography, indocyanine green angiography, and optical coherence tomography. CNVs were divided into completely classic CNV or mainly classic CNV. The relationships of the post-operative logarithm of the minimum angle of resolution (logMAR) visual acuity with preoperative logMAR visual acuity, the shortest distance from the center of the foveal avascular zone to the CNV margin, CNV size, and age were analyzed. RESULTS: Final logMAR visual acuity was improved (defined as a logMAR visual acuity increase of 0.2 or more) in 78 eyes (54%), stable in 47 (32%), and worsened in 21 (14%). Stepwise regression identified CNV size as a significant factor influencing final logMAR visual acuity (R2 = 0.213, p<0.0001), while preoperative logMAR visual acuity, shortest distance from the center of the foveal avascular zone to the CNV margin, and age showed no significant correlation with final logMAR visual acuity. Surgical complications included retinal detachment in six eyes (4%), subretinal hematoma in four eyes (2%), macular hole in three (2%), and proliferative vitreoretinopathy in two (1%). CNV recurred postoperatively in 18 eyes (12%). In 92 eyes with completely classic CNV, visual acuity was improved in 57 (62%), stable in 27 (29%), and worsened in 8 (9%). In 54 eyes with mainly classic CNV, visual acuity was improved in 21(39%), stable in 20 (37%), and worsened in 13 (24%). CONCLUSIONS: Surgical excision of CNV for ARMD was effective for completely classic CNV, and better postoperative visual acuity was achieved in cases of small CNV. Given the fact that photodynamic therapy (PDT) has only been used in Japan since 2004, future study should compare PDT and surgical excision in Japanese subjects for relative merits against surgical risk and postoperative complications, to define indications for PDT and surgical excision. PMID- 16703549 TI - Peribulbar and retrobulbar combined anesthesia for vitreoretinal surgery using ropivacaine. AB - PURPOSE: To evaluate the efficacy and clinical effects of local retrobulbar anesthesia using ropivacaine in vitreoretinal surgery. METHODS: Prospective study. A total of 919 vitreoretinal operations were followed. The operations were divided into three groups, depending on the degree of anesthesia needed. Group A: Vitrectomies with episcleral procedures (208 vitrectomies for detached retina or perforating trauma). Group B: Episcleral procedures only (410 operations for detached retina without vitrectomy). Group C: Vitrectomies without episcleral surgery (301 operations for macular pucker or hole, proliferative diabetic retinopathy, or silicone oil removal). Anesthesia was administered using a 23 gauge Atkinson-type retrobulbar needle, after topical anesthesia. Six mL of the solution containing 7.5 mg ropivacaine/mL were injected into the peribulbar space, and the other 4 mL deeper, into the retrobulbar space. The degree of infiltration of the palpebral region, the motor block in the extrinsic ocular muscles, and pain felt were checked and rated. RESULTS: Swelling of lids was seen in 885 patients (96%); in 21 (2%) swelling was partial. In 13 patients (1%) there were no signs of infiltration. The motor block was total in 801 (87%) eyes, while 118 (12%) had reduced ocular movements. The degree of anesthesia was as follows, considering the three groups together: no pain = 855 (93%) patients; moderate pain = 44 (4%) patients; very strong pain = 20 (2%) patients. No adverse events or side effects were observed. CONCLUSIONS: Ropivacaine used for retrobulbar peribulbar combined anesthesia in vitreoretinal surgery showed excellent clinical efficacy as regards analgesia and muscle akinesia. PMID- 16703550 TI - Did the spectrum of endophthalmitis change? A study comprising 13 years experience with operative therapy of endophthalmitis. AB - PURPOSE: The authors wondered whether the spectrum of endophthalmitis and the type of therapy had changed. METHODS: Files of patients who were operated upon for endophthalmitis between 1988 and 2000 were retrospectively analyzed. They were divided into Group 1 (operated upon 1988 to 1994) and Group 2 (1995 to 2000). RESULTS: Group 1 consisted of 83 patients (43.4% female, mean age 63.9 years), Group 2 of 108 (38.9% female, mean age 64.6 years). Bilateral endophthalmitis occurred in 8.4% of Group 1 patients (3.7% of Group 2 patients). Patients in both groups took on average 1.2 drug types against various internal diseases. The mean interval between first symptoms and presentation in the clinic was 45.7 days in Group 1 (19 days in Group 2; difference statistically significant). There were 63% (Group 1) (70% [Group 2]) cases of postoperative endophthalmitis, among them 58% (Group 1) (63% [Group 2]) after cataract extraction, 6% (Group 1) (5% [Group 2]) after glaucoma surgery, 20% (Group 1) (17% [Group 2]) endogenous and 17% (Group 1) (13% [Group 2]) post traumatic. In Group 2 slightly more Gram-negative bacteria were found. As an initial procedure the following were performed: vitrectomy (70% [Group 1], 88% [Group 2]), removal of crystalline lens (11% [Group 1], 10% [Group 2]), removal of pseudophakos (2% [Group 1], 12% [Group 2]), opening of posterior capsule (1% [Group 1], 9% [Group 2]), and anterior chamber irrigation (36% [Group 1], 43% [Group 2]), often combining procedures. There were significantly more vitrectomies and openings of the posterior capsule in Group 2. Neither the spectrum of secondary and tertiary procedures nor the reasons for such surgery differed in both groups. Neither visual acuity at initial presentation (0.1) nor at final follow-up (0.3) differed between the two groups. The rate of enucleation was less in Group 2 (6% versus 11%) although not statistically significantly. CONCLUSIONS: In Group 2 there were slightly more Gram-negative bacteria and the time interval between initial symptoms and presentation in the clinic had decreased. This can be interpreted as an increase in the severity of the endophthalmitis cases. The final visual acuity was identical in both groups, the enucleation rate improved. PMID- 16703552 TI - Rufloxacin eyedrops: effect of different formulations on ocular pharmacokinetics in rabbits. AB - PURPOSE: To evaluate the aqueous humor pharmacokinetics of rufloxacin in rabbits after topical administration of different formulations, and to individuate the ones showing the best pharmacokinetic profile. METHODS: Six formulations were instilled in rabbit eyes: two pH 7.2 suspensions of non-salified rufloxacin base, or zwitterion (RUF), one of which was viscosized with tamarind seed polysaccharide (TSP); two pH 7.2 solutions of RUF obtained using hydroxypropyl beta-cyclodextrin (CD), one of which was viscosized with TSP; and two pH 5.0 solutions of rufloxacin hydrochloride (RUF-HCl ), one of which was viscosized with TSP. At different times after administration, samples of aqueous humor were withdrawn and analyzed by high-pressure liquid chromatography. The main pharmacokinetic parameters of RUF in the aqueous humor produced by the different formulations were calculated and statistical differences were assessed. RESULTS: The best results, in terms of aqueous humor bioavailability, were observed with two TSP-viscosized formulations: a solution of the hydrochloride (TSP/RUF-HCl) and a suspension of the base (TSP/RUF), followed by the non-viscosized solution of RUF-HCl. The formulations containing CD-solubilized RUF were much less effective. CONCLUSIONS: The present data confirm the significant availability enhancing properties of tamarind seed polysaccharide, and indicate that solubilization of RUF with hydroxypropyl-beta-cyclodextrin (CD/RUF) results in decreased drug availability with respect to standard formulations. Two of the TSP viscosized formulations (RUF suspension and RUF-HCl solution) produced aqueous humor RUF concentrations in the range of activity against Enterobacteriaceae and Pseudomonas aeruginosa, thus warranting further studies on applications of rufloxacin in ocular therapy. PMID- 16703551 TI - Comparison of the effect of Healon, Healon GV, Healon 5, Viscoat, and Ocucoat on platelet aggregation under in vitro conditions. AB - PURPOSE: To study the effect of Healon, Healon GV, Healon 5, Viscoat, and OcuCoat on platelet aggregation under in vitro conditions. METHODS: Ocular viscoelastic devices including Healon, Healon GV, Healon 5, Viscoat, and OcuCoat were studied to investigate the effect of these agents on platelet aggregation under in vitro conditions. The experiments were performed by using platelet-rich plasma with an aggregometer. Aggregation was induced with three different agonists including 5'' adeno-sinediphosphate (ADP), epinephrine (EPI), and collagen (Col ). The results were obtained as a percentage of maximal aggregation and compared with controls using one-way analysis of variance (ANOVA) test. RESULTS: The tests with ADP as aggregating agent revealed that the percentages of maximal aggregation were a mean of 75 +/- 4.35% for ADP only, 67 +/- 4.35% for Healon, 59.33 +/- 3.51% for Healon GV, 70 +/- 3% for Healon 5, 58 +/- 3.46% for Viscoat, and 64 +/- 2% for OcuCoat. Kruskal-Wallis one-way ANOVA test revealed no significant decrease in the percentage of maximal aggregation for all tested substances. With EPI, aggregation was induced in all control samples with a mean of 80.66 +/- 2.08%. The mean percentage of maximal aggregation was 67 +/- 3% for Healon, 77.66 +/- 4.04% for Healon GV, 77 +/- 4% for Healon 5, 80.6 +/- 4.04% for Viscoat, and 65 +/- 5% for OcuCoat. Statistical analysis showed no significant difference. With collagen, maximum aggregation was 74 +/- 5.29% for controls, 65 +/- 4.35% for Healon, 54 +/- 2% for Healon GV, 51 +/- 2.64% for Healon 5, 59 +/- 2% for Viscoat, and 72.66 +/- 1.52% for OcuCoat. Kruskal -Wallis one-way ANOVA test revealed no significant change in the percentage of maximal aggregation for all tested substances in the experiments. CONCLUSIONS: Ophthalmic viscosurgical devices like Healon, Healon GV, Healon 5, and Viscoat that contain glycosaminoglycans and OcuCoat that contains hydroxypropyl methylcellulose have inhibitory effects on platelet aggregation but the effect is not statistically significant and there is no difference among the ocular viscoelastic devices in regard to inhibitory effect on platelet aggregation. PMID- 16703553 TI - Choroidopathy with serous retinal detachment in a patient with polymyositis. AB - PURPOSE: The authors report a unique case of multifocal choroidopathy in association with polymyositis. METHODS: The clinical presentation, laser treatment, and subsequent change in ocular findings are described. RESULTS: A 65 year-old Japanese man with polymyositis experienced mid acute vision impairment in both eyes during systemic corticosteroid treatment for interstitial pneumonia. Fundus examination revealed a gray-white subretinal exudate with serous retinal detachment. Angiographic examination disclosed partial dilatation of the choroidal vein at the posterior pole and dye leakage points in the retinal pigment epithelium. The patient's left eye developed bullous retinal detachment and multi focal choroidal nodules at the posterior pole during the period of tapering of oral corticosteroid administration. Retinal burns were applied to the dye leakage points in the right eye using an argon laser. Vision in the right eye showed rapid recovery due to regression of the serous retinal detachment. However, in the left eye without retinal burns visual acuity decreased to light perception only even after regression of the bullous retinal detachment and choroidopathy. CONCLUSIONS: In serous retinal detachment with choroidopathy associated with polymyositis, laser photocoagulation at leakage points may be useful for rapid restoration of vision and for the prevention of permanent severe vision loss. PMID- 16703554 TI - Photodynamic therapy in retinal angiomatous proliferation stage I. AB - PURPOSE: To report a case of photodynamic therapy (PDT) treatment in an 85-year old patient with retinal angiomatous proliferation (RAP) stage I. METHODS: According to Treatment of Age-Related Macular Degeneration with Photodynamic Therapy and Verteporfin in Photodynamic Therapy study guidelines, two sessions of PDT with verteporfin were performed, which was activated by a diode laser light at 690 nm. RESULTS: The left eye was treated with PDT because of RAP stage I. Even in the early stage of RAP, PDT treatment did not alter the natural course of the disease. In particular, the lesion evolved towards stage III, being initially in stage I, with the final result of development of retinal pigment epithelial (RPE) tear after the second session of PDT treatment. CONCLUSIONS; Even in the early stage of RAP, PDT treatment did not alter the natural course of the disease, with the final result of RPE tear after the second session of PDT treatment. PMID- 16703555 TI - Vitrectomy and silicone oil tamponade for serous macular detachment associated with an optic disk pit. AB - PURPOSE: Description of the results of vitrectomy and silicone oil tamponade to repair serous macular detachment associated with optic disk pit. DESIGN: Interventional case report. METHODS: The authors report on a 17-year-old girl with serous macular detachment with a very thin layer near the fovea which clinically simulated a lamellar macular hole and macular schisis associated with an optic disk pit. The patient underwent pars plana vitrectomy with posterior hyaloid removal and silicone oil tamponade. RESULTS: Good anatomic and functional results were obtained after 1 year of follow-up. CONCLUSIONS: This surgical approach may be a valuable technique for the surgical management of serous macular detachment associated with an optic disk pit. PMID- 16703556 TI - Occurrence of full-thickness macular hole complicating Stargardt disease with ABCR mutation. AB - PURPOSE: To report an unusual episode of full-thickness macular hole complicating Stargardt disease with an ABCR mutation. METHODS: Case report . RESULTS: Fundus examination of a 20-year-old healthy man showed typical fundus manifestation with yellowish-round or fish-like flecks associated with vitreous macular adhesion and a round punched-out area in the right eye. Optical coherence tomography (OCT) illustrated a full-thickness macular hole. Molecular genetic examination of the ABCR gene showed two heterozygous missense mutations: R1108C (CGC-->TGC) in exon 22 and a splicing mutation IVS6--> 1GT - described in the literature in association with Stargardt disease. CONCLUSIONS: Macular hole was once described in other inherited retinal degenerations (Best disease and Bietti crystal line retinopathy). The pathogenesis gives rise to a host of speculations: widespread alteration of the retinal pigment epithelium; inflammatory mechanisms; a minor trauma which might cause subretinal fibrosis. Surgical procedures were not performed on our patient after his ophthalmologic history and findings were considered. PMID- 16703557 TI - Severe orbitopalpebral emphysema after nose blowing requiring emergency decompression. AB - PURPOSE: Orbital emphysema is a frequent condition associated with medial wall fractures after blunt trauma. It is usually a benign, self-limited phenomenon. The authors are unaware of previous reports of severe orbital emphysema needing emergency decompression with no evidence of any previous significant trauma such as the case presented herein. METHODS/RESULTS: A 51-year-old woman had painful swelling of the left orbital region that prevented her from opening her eyelids. She complained of a coincident cold with abundant mucous rhinorrhea. She had blown her nose vigorously several times. A computed tomography scan revealed a left proptosis, an extensive orbitopalpebral emphysema, and a blowout fracture of the medial wall of the left orbit. A complete ophthalmic examination was only possible after a needle decompression. No significant ocular damage was observed, so outpatient treatment was provided. Twenty-eight hours later, the swelling had almost disappeared, and the ocular assessment was normal. CONCLUSIONS: A forceful expiratory effort raising intranasal pressure may cause a medial wall orbital fracture. If the airway hyperpressure episodes are repeated, a severe orbitopalpebral emphysema may develop. Should there be any suspicion of vascular compression, it must be drained to allow the assessment of visual function and theoretically prevent a potentially irreversible ischemic visual loss. PMID- 16703558 TI - Endogenous endophthalmitis by Propionibacterium acnes associated with leflunomide and adalimumab therapy. AB - PURPOSE: To report a case of unusual endogenous endophthalmitis associated with the use of leflunomide and adalimumab. METHODS: A 48-year-old woman on treatment with leflunomide and adalimumab for rheumatoid arthritis developed an endogenous endophthalmitis caused by Propionibacterium acnes. Diagnosis was confirmed by polymerase chain reaction and positive cultures. The patient underwent surgical treatment and intravitreal vancomycin, but the eye developed retinal fibrosis and untreatable retinal detachment. RESULTS: This report of endogenous endophthalmitis associated with the use of anti-tumor necrosis factor alpha (anti -TNF-a) drugs is consistent with those in the literature. P. acnes may induce pathologic reactions in compromised patients and cause endophthalmitis, but only after ocular surgery or in intravenous drug users. The Naranjo probability scale indicated a probable relationship between the drugs and the infection. CONCLUSIONS: Awareness of atypical infectious conditions in patients on anti-TNF a drugs is critical for early diagnosis and good outcome. PMID- 16703559 TI - Hyperacute unilateral gonococcal endophthalmitis in an HIV-infected man without genital infection. AB - PURPOSE: To demonstrate the necessity of obtaining an accurate history from patients presenting abnormal evolution of ophthalmologic diseases. METHODS: A 42 year-old patient, denying any previous ocular or systemic morbidity, presented with an unusual severe and hyperacute gonococcal endophthalmitis with corneal abscess but no concurrent genitourinary infection. Only after a further interview did the patient reveal his human immunodeficiency virus status and a previous diagnosis of acquired immunodeficiency syndrome. RESULTS: Adequate topical and intravenous antibiotic treatment and surgery led to salvage of the eye. CONCLUSIONS: An accurate history should be obtained by patients with an abnormal course of an ophthalmologic disease, focusing on immunologic deficiencies that can cause extremely serious ophthalmologic complications with ensuing risk of visual impairment or ocular loss (bulbar enucleation). PMID- 16703560 TI - Retinocytoma associated with calcified vitreous deposits. AB - PURPOSE: To report a case of bilateral retinocytoma associated with calcified vitreous deposits. METHODS: Case report. RESULTS: On routine examination, a 35 year-old asymptomatic father of a child with bilateral retinoblastoma presented bilateral retinocytoma associated with vitreous calcifications, in the vicinity of the retinocytoma in his left eye. Fundus photographic documentation and fluorescein angiography were performed. The patient has been followed up for 10 years. CONCLUSIONS: The lesions in both eyes have remained stable without signs of growth or malignant transformation. Calcified vitreous deposits are a recently described feature of retinocytoma in addition to the three classic features: translucent retinal mass, retinal pigment epithelial alteration, and calcification. PMID- 16703562 TI - Use of high-frequency ultrasound for follow-up and visualization of intracanalicular plugs. PMID- 16703561 TI - Optical coherence tomography findings in branch retinal artery occlusion. AB - PURPOSE: To report the optical coherence tomography (OCT) findings in a 35-year old man with acute inferior branch retinal artery occlusion. METHODS: OCT findings in acute branch retinal artery occlusion were evaluated. RESULTS: OCT disclosed diffuse thickening of the neurosensory retina in the inferior perifoveolar area. Increased reflectivity was noted in the inner retinal layers rom the surface of the retina to the photoreceptor layers. Decreased reflectivity was observed from the photoreceptor layers and the retinal pigment epithelium secondary to the shadowing effect. Foveolar depression, photoreceptor layer in the fovae, and retinal pigment epithelium underneath the fovea were normal. OCT findings in the superior perifoveolar area were also normal. CONCLUSIONS: In the retinal artery occlusion, denaturation and breakdown of the normally transparent intracellular protein and an increase in the intracellular fluid cause ischemic whitening of the retina. Otherwise there is no retinal thickening secondary to the accumulation of serous fluid escaping from retinal capillaries into the extracellular space. The OCT findings support these descriptions. PMID- 16703564 TI - Sustained B7/CD28 interactions and resultant phosphatidylinositol 3-kinase activity maintain G1-->S phase transitions at an optimal rate. AB - Twenty-four hours of TCR engagement and CD28 costimulation was found sufficient to elicit an optimal rate of cell division over a 72-h period only when a high concentration of IL-2 was produced in the culture and remained readily available to the CD4(+) T cells. The cell division response could be aborted following 24 h of stimulation by the simultaneous abrogation of IL-2R signaling and the blockade of CD28 or TCR ligands. Biochemical and pharmacologic studies indicated that a phosphatidylinositol 3-kinase-Akt signaling cascade costimulated by the TCR and CD28 maintained the blasting cell division rate at a maximal level beyond 24 h even when IL-2 was withdrawn, neutralized, or exhausted. These data show that CD4(+) T cells remain sensitive to antigens (Ag) and costimulatory signals throughout the clonal expansion response. Furthermore, only those T cells that perceive the presence of a continued threat in the form of Ag/MHC complexes and B7 costimulatory ligands or a high concentration of a growth factor are directed to remain in cell cycle. PMID- 16703563 TI - Histamine modulates mast cell degranulation through an indirect mechanism in a model IgE-mediated reaction. AB - Histamine is released in inflammatory reactions and exerts an immunoregulatory function on cells present in the microenvironment. In this study, we compared the effect of histamine on degranulation of mast cells derived from animals bearing a parasitic infection with those from uninfected animals. Peritoneal mast cells (PMC) were obtained 24 days after infection of Wistar rats with Toxocara canis. The degree of degranulation was assessed either morphologically or by measuring the release of beta-hexosaminidase and TNF-alpha. Non-purified PMC or mast cells immunomagnetically purified with mAb AA4 were used. An increase in degranulation of non-purified mast cells from infected animals was observed after incubation with histamine in vitro or when histamine was injected into the peritoneal cavity. When a purified mast cell population was used, this effect was no longer observed. Supernatants from spleen cells stimulated with histamine induced degranulation of purified mast cells, and again, this was potentiated with PMC from infected animals. However, when supernatants from peritoneal macrophages similarly stimulated were used, a reduction in the degranulation of PMC from infected animals was observed. Our results suggest that histamine may act as a regulator of mast cell degranulation, thus modulating inflammatory responses due to infection with certain parasites. PMID- 16703565 TI - Revisiting the specificity of the MHC class II transactivator CIITA in vivo. AB - CIITA is a master regulatory factor for the expression of MHC class II (MHC-II) and accessory genes involved in Ag presentation. It has recently been suggested that CIITA also regulates numerous other genes having diverse functions within and outside the immune system. To determine whether these genes are indeed relevant targets of CIITA in vivo, we studied their expression in CIITA transgenic and CIITA-deficient mice. In contrast to the decisive control of MHC II and related genes by CIITA, nine putative non-MHC target genes (Eif3s2, Kpna6, Tap1, Yars, Col1a2, Ctse, Ptprr, Tnfsf6 and Plxna1) were found to be CIITA independent in all cell types examined. Two other target genes, encoding IL-4 and IFN-gamma, were indeed found to be up- and down-regulated, respectively, in CIITA transgenic CD4(+) T cells. However, there was no correlation between MHC-II expression and this Th2 bias at the level of individual transgenic T cells, indicating an indirect control by CIITA. These results show that MHC-II restricted Ag presentation, and its indirect influences on T cells, remains the only pathway under direct control by CIITA in vivo. They also imply that precisely regulated MHC-II expression is essential for maintaining a proper Th1 Th2 balance. PMID- 16703566 TI - Macrophages require distinct arginine catabolism and transport systems for proliferation and for activation. AB - In murine macrophages, as a result of arginine catabolism during activation, citruline is produced under the effect of IFN-gamma and LPS, and ornithine and polyamines by IL-4 and IL-10. For proliferation, arginine is required from the extracellular medium and is used for protein synthesis. During activation, most arginine (>95% in 6 h) was metabolized, while under proliferation only half was incorporated into proteins. Under basal conditions, this amino acid was preferentially transported by y(+)L activity. During activation, arginine transport increased drastically (4-5-fold) through y(+) cationic amino acid transporter (CAT) activity. By contrast, M-CSF induced only a modest increase in uptake (0.5-fold). The increase in arginine transport during activation, but not proliferation, was mediated by the SLC7A2/Cat2 gene. SLC7A1/Cat1 is constitutively expressed, and is not modified by proliferating or activating agents. M-CSF-dependent proliferation was not affected in the macrophages of SLC7A2 knockout mice; however, these cells showed a drastic reduction in the production of citruline or ornithine and polyamines during activation. The data show that a large increase in a specific transport system (CAT2) is necessary for activation-induced arginine metabolism, while arginine is in excess for the requirements of proliferation and a modest increase in transport occurs. PMID- 16703568 TI - An absence of reactive oxygen species improves the resolution of lung influenza infection. AB - Three influenza virus pandemics occurred in the last century, in 1918 killing 40 50 million people. In the absence of strain-specific vaccines, with potential resistance to antivirals and the threat of an imminent pandemic, strategies that alleviate symptoms are a priority. Reactive oxygen species are potent antimicrobial agents but cause immunopathology when produced in excess. Mice lacking a functional phagocyte NADPH oxidase (Cybb tm1 mice) or treated with the metalloporphyrin antioxidant manganese (III) tetrakis (N-ethyl pyridinium-2-yl) porpyhrin (MnTE-2-PyP) show heightened inflammatory infiltrates in their airways in response to pulmonary influenza infection, with augmented macrophage populations and a Th1-skewed T cell infiltrate. Underlying this exuberant macrophage response was a significant reduction in apoptosis and down-regulation of the myeloid inhibitory molecule CD200. Both, Cybb tm1 and MnTE-2-PyP-treated mice exhibited a reduced influenza titer in the lung parenchyma. Inflammatory infiltrate into the lung parenchyma was markedly reduced and lung function significantly improved. Manipulation of the homeostatic control of myeloid cells by inflammatory mediators therefore represents a novel therapeutic strategy in the treatment of influenza virus infection. PMID- 16703567 TI - TLR4, TLR9 and MyD88 are not required for the positive selection of autoreactive B cells into the primary repertoire. AB - Toll-like receptors (TLR) have been shown to play an essential role in the generation of autoantibodies in mouse models of autoimmunity, but the timing and context of these effects are poorly understood. One hypothesis is that TLR ligands assist in the positive selection of self-reactive B cells into the primary repertoire and, in this way, distinguish between immunogenic and tolerogenic forms of self-antigen. To explore this idea we generated hen egg lysozyme-specific immunoglobulin (Ig(HEL)) and isotype class-switching anti-HEL mice deficient in MyD88, TLR4 or TLR9 signalling and studied B cell development and autoantibody secretion in the presence or absence of an intracellular form of self-antigen HEL that positively selects B1 cells. Our findings show that TLR4, TLR9 and MyD88 are not required for the positive selection of autoreactive B cells in the primary B cell repertoire, nor is MyD88 required for the generation of isotype-switched antibodies in the absence of antigen. These results suggest that the significant effects of TLR on autoimmunity occur in the established repertoire and not during B cell development. PMID- 16703570 TI - Macroporous polyacrylamide monolithic gels with immobilized metal affinity ligands: the effect of porous structure and ligand coupling chemistry on protein binding. AB - Macroporous polyacrylamide gels (MPAAG) with iminodiacetic acid (IDA) functionality were prepared by (i) chemical modification of polyacrylamide gel, (ii) co-polymerization of acrylamide with allyl glycidyl ether (AGE) and N,N'metylene-bis(acrylamide) (MBAAm) followed by coupling IDA ligand or (iii) by copolymerization of acrylamide and MBAAm with functional monomer carrying IDA functionality (1-(N,N-bis(carboxymethyl)amino-3-allylglycerol). Screening for optimized conditions for the production of the MPAAG with required porous properties was performed in a 96-well chromatographic format that allowed parallel production and analysis of the MPAAG prepared from reaction mixtures with different compositions. Scanning electron microscopy of the fabricated MPAAG revealed two different types of the porous structures: monomodal macroporous structure with large interconnected pores separated by dense non-porous pore walls in case of plain gels or gels produced via copolymerization with AGE. The other type of the MPAAG (gel produced via co-polymerization with functional monomer carrying IDA-functionality) had bimodal pore structure with large interconnected pores separated by the pore walls pierced through with micropores. The effect of different modifications of MPAAG monoliths and of porous structure of the MPAAG (monomodal and bimodal porous structure) on protein binding has been evaluated. PMID- 16703569 TI - Ion-exchange macroporous hydrophilic gel monolith with grafted polymer brushes. AB - The grafting of functional polymers to the pore surface of macroporous monolithic polyacrylamide cryogels was found to be an efficient and convenient method for the preparation of macroporous polyacrylamide gels, so-called cryogels (pAAm cryogels), with both controlled extent of functional group incorporated and with tailored surface chemistries. Anion-exchange polymer chains of poly(2 (dimethylamino)ethyl methacrylate) (pDMAEMA) and poly([2-(methacryloyloxy)ethyl] trimethylammonium chloride) (pMETA), and cation-exchange polymer chains of polyacrylate have been grafted onto pAAm cryogels using potassium diperiodatocuprate as initiator. It was possible to achieve the ion-exchange capacity up to 0.2-0.5 mmol/ml. The graft polymerization did not alter the macroporous structure of the pAAm cryogel, however the flow rate of solutes through the cryogel matrix decreased with increase in the density of polymer grafted. The sorption of low-molecular-weight (metal ion, dye) and high-molecular weight (protein) substances on the grafted monolithic pAAm column has been studied. The results indicate that a 'tentacle'-type binding of protein to grafted polymer depended on the architecture of the grafted polymer layer and took place after a certain degree of grafting has been reached. The binding of proteins by tentacle-like polymer chains allowed for increasing the binding capacity for proteins on the grafted pAAm cryogels up to 6-12 mg/ml. PMID- 16703571 TI - Role of chemical structure in molecular recognition by transferrin. AB - Studies of molecular recognition of chiral compounds by proteins are of importance from many points of view. The biological role of proteins in their interaction with small molecules is of fundamental interest and can be used in many different fields, for instance for in vitro analysis of optically active compounds. Studies in these areas need a detailed study of the interaction sites on the protein surface and the relationship between chemical structure and the complex formation ability of small molecules, such as drugs. The electrophoretic migration of charged compounds through a protein zone may provide information about the surface properties of the macromolecule in the interaction site. The interaction of human serum transferrin with tryptophan-methyl- (TME), ethyl- (TEE) and butyl-esters (TBE) has been investigated by capillary electrophoresis (CE) and model calculations. Differences in the separation of tryptophan derivatives were obtained by varying experimental parameters such as, pH, ionic strength of background electrolyte and the length of transferrin zone. Limited separation of the enantiomer pairs were observed at pH 5 and 7 with a maximum resolution at pH 6. The size of the ligands coupled to the chiral centre has importance in stereoselective recognition; however, a direct comparison of resolution different in same runs may lead to false conclusion if the experimental conditions are not comparable. With a careful evaluation of the data we obtained significant differences between the resolution of the smallest enantiomer pair compared to those of tryptophan derivatives with longer alkyl chains. PMID- 16703572 TI - Limitations of tidal breathing parameters in clinical trials with young children. PMID- 16703573 TI - Six-minute walk test in children and adolescents with cystic fibrosis. AB - The 6-min walk test is a simple, rapid, and low-cost method that determines tolerance to exercise. We examined the reproducibility of the 6-min walk test in 16 children with cystic fibrosis (11 female, 5 male; age range, 11.0 +/- 1.9 years). We related the distance walked and the work performed (distance walked x body weight) with nutritional (body mass index and respiratory muscle strength) and clinical (degree of bronchial obstruction and Shwachman score) status. Patients were asked to walk as far as possible upon verbal command on two occasions. There was no statistical difference between distances walked (582.3 +/ 60 and 598.2 +/- 56.8 m, P = 0.31), heart rate, respiratory rate, pulse oxygen saturation, arterial blood pressure, dyspnea, and percentage of maximal heart rate for age in the two tests. Distance walked correlated (Pearson) with maximal expiratory pressure (98.6 +/- 28.1 cmH2O, r = 0.60, P < 0.01), maximal heart rate (157.9 +/- 10.1 bpm, r = 0.59, P < 0.02), Borg dyspnea scale (1.7 +/- 2.4, r = 0.55, P < 0.03), and double product (blood pressure x heart rate; r = 0.59, P < 0.02). The product of distance walked and body weight (work) correlated (Pearson) with height (r = 0.83, P = 0.000), maximal expiratory pressure (r = 0.64, P < 0.01), systolic blood pressure (r = 0.56, P < 0.02), and diastolic blood pressure (r = 0.55, P < 0.03). We conclude that the 6-min walk test is reproducible and easy to perform in children and adolescents with cystic fibrosis. The distance walked was related to the clinical variables studied. Work in the 6-min walk test may be an additional parameter in the determination of physical capacity. PMID- 16703574 TI - Necrotizing pneumococcal pneumonia in children: the role of pulmonary gangrene. AB - Little is known about the mechanism of necrotizing pneumonia caused by Streptococcus pneumoniae in children. Pulmonary gangrene secondary to vascular thrombosis was reported in adults with necrotizing pneumococcal pneumonia. We conducted a retrospective study of 15 children with a diagnosis of necrotizing pneumococcal pneumonia at National Taiwan University Hospital to explore its association with pulmonary gangrene, based on evidence from chest computed tomography, serial chest radiographic patterns, and pathologic results. S. pneumoniae serotype 14 was the prevalent pneumococcal serotype. Overall, 63.6% of isolates were not susceptible to penicillin. One child with pneumonia caused by S. pneumoniae serotype 3 complicated by hemolytic uremic syndrome had a rapidly fatal course. An autopsy in this patient documented lung necrosis and pulmonary gangrene. Radiographic follow-up was performed during the clinical course in 9 patients, and showed no evidence of pulmonary gangrene. Four children had no radiographic follow-up. The relationship between pulmonary gangrene and necrotizing pneumonia was unclear in the remaining one. In conclusion, necrotizing pneumococcal pneumonia may be infrequently associated with pulmonary gangrene in children. PMID- 16703575 TI - Zinc status in infantile wheezing. AB - The increase in prevalence of asthma is strongly dependent on environmental factors, including diet. Significant decreases in the intake of dietary zinc may be an important contributing factor to the increasing incidence of wheezing and asthma, but there have been no studies evaluating zinc levels in wheezy infants. Our objective was to investigate the zinc status of wheezy infants. Wheezy infants (n = 34) and healthy children (n = 14) were included in the study. Total IgE and eosinophil counts were obtained, and skin testing was done with a battery of 25 antigens with appropriate positive and negative controls. Levels of zinc were determined in hair, using a Polarized Zeeman Atomic Absorption Spectrophotometer (Hitachi Z-800). No significant difference was observed in peripheral blood eosinophil counts and total IgE levels among groups (P > 0.05). Hair zinc levels were significantly lower in wheezy infants (P < 0.001). In conclusion, hair zinc levels were lower in wheezy infants than in healthy controls, suggesting that zinc deficiency may influence the risk of wheezing in early childhood. PMID- 16703576 TI - Exhaled nitric oxide in healthy nonatopic school-age children: determinants and height-adjusted reference values. AB - Exhaled nitric oxide (FENO) was proposed as a marker of airway inflammation, but data about FENO in healthy children measured with standardized methods are so far limited. In order to assess the determinants of FENO in healthy children, we investigated a population-based sample of school-age children (n = 276) with a questionnaire, skin-prick tests, spirometry, and the measurement of FENO. The FENO of 114 nonatopic and nonsmoking children considered healthy were analyzed with stepwise multiple regression analysis, which showed significant associations with age, standing height, weight, and body surface area, but not with gender. Height was found to be the best independent variable for the regression equation for FENO, which on average showed an increase in the height range of 120-180 cm from 7 to 14 ppb. In the random sample of children, increased FENO was associated with atopy (odds ratio, 9.0; 95% confidence interval, 3.9-21.1; P < 0.0001), and significantly with allergic rhinitis and atopic dermatitis, but not with asthma. Respiratory symptom-free children with skin-prick test positivity had significantly higher FENO than healthy nonatopic subjects. We conclude that height is the best determinant of FENO in healthy children. Due to the strong effect of atopy, FENO data should not be interpreted without knowing the atopic status of the child. The present reference values of FENO may serve in clinical assessments for measuring airway inflammation in children. PMID- 16703577 TI - Association of birth weight with asthma-related outcomes at age 2 years. AB - BACKGROUND: Although lower birth weight associated with prematurity raises the risk of asthma in childhood, few prospective studies have examined higher birth weight, and few have separated the two components of birth weight, fetal growth and length of gestation. OBJECTIVE: To examine the associations of fetal growth and length of gestation with asthma-related outcomes by age 2 years. METHODS: We studied 1,372 infants and toddlers born after 34 weeks' gestation in Project Viva, a prospective cohort study of pregnant mothers and their children. The main outcome measures were parent report of (1) any wheezing (or whistling in the chest) from birth to age 2 years, (2) recurrent wheezing during the first 2 years of life, and (3) doctor's diagnosis of asthma, wheeze or reactive airway disease ("asthma") by age 2. We calculated gestational age from the last menstrual period or ultrasound examination, and determined birth weight for gestational age z value ("fetal growth") using US national reference data. RESULTS: Infants' mean birth weight was 3,527 (SD, 517; range, 1,559-5,528) grams. By age 2 years, 34% of children had any wheezing, 14% had recurrent wheezing, and 16% had doctor diagnosed asthma. After adjusting for several parent, child, and household characteristics in logistic regression models, we found that infants with birth weight > or = 4,000 g were not more likely to have any wheezing (odds ratio (OR), 0.91; 95% confidence interval (CI): 0.62, 1.34) or doctor-diagnosed asthma (OR, 0.80; 95% CI: 0.49, 1.31) than infants with birth weight 3,500-3,999 g. In models examining length of gestation and fetal growth separately, neither the highest nor the lowest groups of either predictor were associated with the three outcomes. Boys had a higher incidence of asthma-related outcomes than girls, and exposure to passive smoking, parental history of asthma, and exposure to older siblings were all associated with greater risk of recurrent wheeze or asthma related outcomes at age 2 years. CONCLUSION: Although male sex, exposure to smoking, parental history of asthma, and exposure to older siblings were associated with increased risk of wheezing and asthma-related outcomes in this prospective study of children born after 34 weeks gestation, fetal growth and length of gestation were not. PMID- 16703578 TI - Lack of association between NOS2 pentanucleotide repeat polymorphism and asthma phenotypes or exhaled nitric oxide concentration. AB - Nitric oxide (NO) plays an immunoregulatory role in balancing cellular immunity. The expression of inducible nitric oxide synthase gene (NOS2) is upregulated upon exposure to proinflammatory cytokines and microbial exposure. The (CCTTT)n polymorphism in NOS2 promoter confers protection against infections and immunological disorders including atopy. We investigated the association between (CCTTT)n and asthma traits in Chinese children. Asthmatic children between 5 and 18 years of age and non-allergic controls were recruited. Plasma total and specific IgEs were measured by immunoassays, and exhaled NO concentration was quantified online by chemiluminescence. NOS2 (CCTTT)n was genotyped by GeneScan analysis. The mean (SD) age of 291 asthmatics and 172 controls were 11.1 (3.8) years and 11.6 (4.0) years, respectively (P = 0.259). NOS2 (CCTTT)n followed Hardy-Weinberg equilibrium in both groups, and its uni-modal allele distribution peaks at 12-repeat. Significant interethnic differences in (CCTTT)n alleles were observed, with our Chinese having less 13-repeat (Pc = 0.022) but more 17-repeat (Pc = 0.033) than Caucasians. The frequency of 14-repeat allele was similar in our Chinese as compared to Japanese (Pc = 0.32). Multivariate regression analyses failed to detect any association between this polymorphic marker and asthma diagnosis (P = 0.949), atopy (P = 0.305), IgE sensitization to aeroallergens (P > 0.2 for all), or FeNO (P = 0.847). These findings do not support NOS2 to be a major candidate gene for asthma or IgE-mediated allergic diseases in Chinese children. PMID- 16703579 TI - Microbiology, safety, and pharmacokinetics of aztreonam lysinate for inhalation in patients with cystic fibrosis. AB - BACKGROUND: Aztreonam lysinate for inhalation (AI) is a novel monobactam formulation being investigated for pulmonary Pseudomonas aeruginosa infections in patients with cystic fibrosis (CF). METHODS: Pre-clinical studies investigated the pre- and post-nebulization activity of AI and its activity in the presence of CF sputum. A double-blind, placebo-controlled, dose-escalation trial determined pharmacokinetics and tolerability of AI in subjects with CF. Single daily escalating doses of AI 75, 150, or 225 mg, or placebo were self-administered using an eFlow Electronic Nebulizer. Sputum samples were collected up to 4 hr and blood samples up to 8 hr post-dose. RESULTS: AI activity against multiple CF isolates was retained after nebulization via eFlow, and activity was not inhibited by CF sputum. All 12 adult subjects and 11/12 adolescents tolerated all AI doses. One patient had an asymptomatic FEV1 decrease > 20% with the 150 mg dose. Median aztreonam sputum concentrations in adults 10 min after AI 75, 150, and 225 mg were 383, 879, and 985 microg/g, respectively. Median sputum concentrations in adolescents 10 min after AI 75, 150, and 225 mg were 324, 387, and 260 microg/g, respectively. Systemic exposure to AI was low. Plasma pharmacokinetics in adults receiving AI 75 mg were Cmax = 419 ng/g, Tmax = 0.99 hr, t1/2 = 2.1 hr. Aztreonam concentrations in sputum were at or above the MIC50 for at least 4 hr post-dose. CONCLUSION: These data support the continued development of AI for treatment of pulmonary infections in patients with CF. PMID- 16703580 TI - Nebulized therapies for childhood pulmonary hypertension: an in vitro model. AB - OBJECTIVES: Sildenafil, tezosentan, and prostacyclin reduce pulmonary vascular pressures in pulmonary hypertension, but have potential to vasodilate the systemic circulation. Nebulized vasodilators allow targeted drug delivery, high local drug concentrations, less systemic hypotension, and better matching of the lung's ventilation and perfusion. We aimed to estimate pulmonary deposition of these drugs from commonly employed nebulizers using in vitro techniques and to create a mathematical model to predict inspired mass of aerosol. DESIGN: Lung deposition was estimated by characterization of drug output and particle size distribution (PSD) of nebulizers using helium-neon laser diffraction techniques. A mathematical model for each device was created to estimate pulmonary deposition using patients' breathing patterns and was verified with a mechanical-breathing model. RESULTS: Total output and PSD were similar for the Hudson Updraft II and Whisperjet nebulizers, consisting of half the nebulizer's charge, with (1/4) of particles < or = 5 microm, in the respirable fraction (RF). Drug output increased with inspiratory flow for the Pari LC Star. Differences were noted in device performance, depending on the drug tested. Estimated pulmonary deposition (mean, 95% CI) was 8.1 (7.2, 9.0)% of the initial drug charge for the Hudson Updraft II, 6.4 (5.8, 7.0)% for the Whisperjet, and 33.0 (28.3, 37.9)% for the Pari LC Star. A mechanical model was consistent with our mathematical model. CONCLUSIONS: All drugs could be nebulized, but expected pulmonary deposition varied depending on the nebulizer and drug. PMID- 16703581 TI - Plasma concentrations of soluble CTLA-4, CD28, CD80 and CD86 costimulatory molecules reflect disease severity of acute asthma in children. AB - Recent studies have demonstrated an elevation of plasma soluble costimulatory molecules B7.1 (CD80) and B7.2 (CD86), and their T lymphocyte counter receptors cytotoxic T lymphocyte associated antigen-4 (CTLA-4) and CD28 in asthmatic patients. We hypothesized that these costimulatory molecules may reflect the severity of asthma and investigated the longitudinal changes of these soluble costimulatory molecules, and their clinical significance in children with an asthmatic exacerbation. Sixteen children hospitalized for asthmatic exacerbation were recruited and treated with systemic corticosteroid (CS) for 5 days. Plasma total Immunoglobulin E (IgE) and eosinophil cationic protein (ECP) concentrations were measured by microparticle immunoassay and fluorescence enzyme immunoassay, respectively. Soluble CTLA-4 (sCTLA-4), sCD28, sCD80, and sCD86 concentrations in plasma were measured by enzyme-linked immunosorbent assay, and their relationships with asthma severity, total IgE, ECP concentrations, and blood eosinophil count were analyzed. Plasma sCTLA-4, sCD28, sCD80, and sCD86 concentrations in patients were highest during the acute attack. They decreased significantly with a parallel increase of peak expiratory flow rate (PEFR) after CS treatment (all P < 0.05). Plasma sCTLA-4, sCD28, and sCD86 concentrations of patients at recruitment were inversely correlated with PEFR, whereas plasma sCD28 and sCD86 concentrations correlated positively with eosinophil count and plasma ECP concentration (all P < 0.05). Although there was no such correlation with IgE concentration, sCD28 was correlated very significantly with eosinophil count (r = 0.83, P < 0.0001). In conclusion, plasma sCTLA-4, sCD28, sCD86, and sCD80 concentrations may reflect the severity of acute asthma and more studies on larger cohorts are needed to assess whether these markers are useful for assessing asthmatic exacerbation in children. PMID- 16703582 TI - Conservative treatment of spontaneous tracheal rupture. AB - Spontaneous tracheobronchial ruptures are uncommon injuries, especially in the pediatric age group. Tracheal injuries, independent of their origin, may be life threatening. Here we present the first report of a 14-year-old boy who presented with subcutaneous emphysema, pneumomediastinum, and pneumothorax on day 3, due to spontaneous posterior tracheal-wall rupture following paroxysmal productive coughing. The diagnosis was established using a computed tomography scan of the chest, and tracheobronchoscopy and esophagoscopy under general anesthesia. He was endotracheally intubated and ventilated in the intensive care unit. Such tracheal defects, bridgeable by an endotracheal tube, may permit conservative treatment. The patient was discharged on day 10, and follow-up revealed no late complications. PMID- 16703583 TI - Primary pulmonary Hodgkin's disease and tuberculosis in an 11-year-old boy: case report and review of the literature. AB - Tuberculosis (TB) has been described in association with different malignancies including Hodgkin's disease. However, the association with primary pulmonary Hodgkin's disease (PPHD) is hardly reported in literature and in teenage is quite exceptional. We report a case of an 11 years old boy in whom the diagnosis of tuberculosis preceded and delayed the diagnosis of PPHL. PMID- 16703584 TI - Superior vena cava syndrome related to indwelling intravenous catheters in patients with cystic fibrosis. AB - Patients with cystic fibrosis (CF) often need long-term implanted vascular-access devices for intravenous antibiotics for chronic lower respiratory tract infections. These devices are not without complications, including infection, occlusion, and vascular thrombosis. Such thrombosis can result in superior vena cava (SVC) syndrome due to the position of the catheter proximal to the right atrium. SVC syndrome in CF patients, however, is rarely reported in the literature, suggesting that its incidence is uncommon. We describe three patients with SVC syndrome as a consequence of implanted vascular-access devices. PMID- 16703585 TI - BglII-based panhandle and reverse panhandle PCR approaches increase capability for cloning der(II) and der(other) genomic breakpoint junctions of MLL translocations. AB - Panhandle PCR techniques to amplify known sequence flanked by unknown sequence have been useful for MLL genomic breakpoint junctions and fusion transcripts because MLL has a large number of partner genes. However, genomic panhandle PCR approaches are impeded when the restriction fragment that contains the breakpoint junction is too large to amplify. We devised new panhandle PCR approaches for MLL genomic breakpoint junctions that create the template from BglII restriction fragments by attaching MLL sequence to a BglII site in the partner gene. This leads to the annealing of MLL and its complement in the handle and creates an intrastrand loop containing the breakpoint junction sequence for amplification with primers all from MLL. BglII panhandle PCR for der(11) breakpoint junctions was accomplished by ligating a phosphorylated oligonucleotide containing a BglII overhang and sequence complementary to MLL exon 7 to the 3' ends of BglII digested DNA, and forming the template from the sense strand of DNA. In BglII reverse panhandle PCR for der(other) breakpoint junctions, a phosphorylated oligonucleotide containing a BglII overhang and the complement of antisense sequence in MLL exon 10 was ligated to the 3' ends of BglII digested DNA, and the template was formed from the antisense strand of DNA. These approaches amplified 5'-MLL-MLLT4-3' and 5'-AFF1-MLL-3' breakpoint junctions. The former is significant because few t(6;11) genomic breakpoint junctions have been sequenced. BglII panhandle PCR approaches increase the possibilities for cloning MLL genomic breakpoint junctions where there is heterogeneity in partner genes and breakpoint locations. PMID- 16703586 TI - Clinical scoring systems in cystic fibrosis. AB - The first cystic fibrosis (CF) scoring system was published in 1958. Since then, many other scoring systems were developed. Clinical parameters, details about statistical evaluations, and recent strategic uses of scores were identified. Several similar scores aiming to assess chronic illness severity (Shwachman Kulczycki score and a modification, Cooperman, Berneze-score and the NIH score) have not been evaluated and are out of date, given the changing natural history of CF. Of the current scoring systems, the modified Shwachman score by Doershuk is perhaps most reliable for describing follow-up studies. Scores designed for acute changes and short-term evaluation were also developed. The modified Huang score may be useful in the prognostic evaluation of patients with end-stage disease. It could also be used for discrimination of adult patients with differing disease severity and for longitudinal evaluation. Scores assessing pulmonary exacerbations could help provide consensus among clinicians regarding the need for intervention. Most of these scores require further evaluation. Although scores could provide an objective measure of disease severity, progression, need for and response to interventions, including value in selecting patients for lung transplantation and as an outcome measure for research studies, no scoring system can fulfill all these objectives. Nevertheless, there is a need for the development of a modern day longitudinal score that is sensitive, valid and reproducible, to reflect the milder disease status of patients. PMID- 16703587 TI - Fracture risk after the diagnosis of Parkinson's disease: Influence of concomitant dementia. AB - In an inception cohort of 196 Olmsted County, Minnesota, residents with Parkinson's disease (PD) first recognized in 1976 to 1995, we tested whether the increased risk of bone fractures is associated with concomitant dementia. Using the data resources of the Rochester Epidemiology Project, information about PD, dementia, other clinical risk factors for fracture and fracture events was obtained from review of complete inpatient and outpatient medical records spanning each subject's residence in the community. Compared to an equal number of age- and sex-matched non-PD referent subjects from the community, PD patients were at a 2.2-fold increased risk of fractures generally and a 3.2-fold greater risk of hip fractures specifically. Adjusting for age, the independent predictors of overall fracture risk in the PD subjects included female sex (hazard ratio [HR] 1.6; 95% confidence interval [CI], 1.1-2.3), dementia (HR, 1.6; 95% CI, 1.1 2.4) and chronic depression, which was associated with a reduced risk (HR, 0.4; 95% CI, 0.2-0.8). Hip fractures were predicted by dementia (HR, 2.2; 95% CI, 1.2 4.1). The increased fracture risk in patients with PD is not entirely explained by concomitant dementia, and additional study is needed to determine the relative contributions to fracture risk of falls versus bone loss in these patients. PMID- 16703588 TI - Dropped head syndrome in Parkinson's disease. AB - We determined the frequency of dropped head syndrome in Parkinson's disease (PD) in Japan and evaluated its clinical correlates. A total of 252 consecutive patients with PD who visited our hospital were studied. Dropped head syndrome was found in 15 patients (6.0%) (3 men, 12 women; mean age at onset of PD, 62.8 +/- 11.5 years). The interval before emergence of dropped head after disease onset was 5.4 +/- 4.3 years (-0.5 to 15 years). The Hoehn-Yahr score at the on stage was 3.2 +/- 0.7; at the off stage 3.5 +/- 0.8. Of those 15 patients, 8 had major symptoms of rigidity and akinesia. In 2 patients, administration of a dopamine agonist appeared to evoke dropped head syndrome. An increase in and/or the addition of antiparkinsonian drugs alleviated head drop in 4 patients and reduced head drop in 7 patients. Any medication was not effective for 4 patients. Dropped head syndrome in PD is not rare in Japan. It is more often observed in women and is associated with patients who primarily suffer rigidity and akinesia. Dropped head syndrome in these patients appears to be produced by disproportionate tonus of the neck muscles. It is modulated by antiparkinsonian drugs and is considered to be a type of dystonia. PMID- 16703589 TI - Effect of vagal nerve stimulation in a case of Tourette's syndrome and complex partial epilepsy. AB - We report on a 30-year-old man with Tourette's syndrome (TS) and medication refractory epilepsy whose tics improved after implantation of a vagal nerve stimulator (VNS). To verify the patient's observation, we performed a blinded video assessment using the modified Rush video-based tic rating scale. The patient underwent two separate video recordings (VNS on and VNS off). A rater, blinded to patient's VNS status, evaluated the videos with the modified Rush video-based tic rating scale. There were improvements in total tic score and motor and phonic tic frequency. If verified by controlled clinical trials, this observation may provide insights into the pathophysiology of tics and may lead to a novel therapy for patients with severe TS. PMID- 16703590 TI - MEP latency shift after implantation of deep brain stimulation systems in the subthalamic nucleus in patients with advanced Parkinson's disease. AB - Deep brain stimulation (DBS) into the subthalamic nucleus (STN) is a highly effective treatment for advanced Parkinson's disease (PD). The consequences of STN stimulation on intracortical and corticospinal excitability have been addressed in a few studies using transcranial magnetic stimulation (TMS). Although excitability measurements were compared between the STN stimulation OFF and ON condition, in these experiments, there are no longitudinal studies examining the impact of electrode implantation per se on motor excitability. Here, we explored the effects of STN electrode implantation on resting motor thresholds (RMT), motor evoked potential (MEP) recruitment curves, and MEP onset latencies on 2 consecutive days before and shortly after STN surgery with the stimulator switched off, thus avoiding the effects of chronic DBS on the motor system, in 8 PD patients not taking any dopaminergic medication. After surgery, RMT and MEP recruitment curves were unchanged. In contrast, MEP onset latencies were significantly shorter when examined in relaxed muscles but were unchanged under preactivation. We hypothesize that postoperatively TMS pulses induced small currents in scalp leads underneath the TMS coil connecting the external stimulator with STN electrodes leading to inadvertent stimulation of fast conducting descending neural elements in the vicinity of the STN, thereby producing submotor threshold descending volleys. These "conditioning" volleys probably preactivated spinal motor neurons leading to earlier suprathreshold activation by the multiple corticospinal volleys produced by TMS of the motor cortex. These TMS effects need to be considered when interpreting results of excitability measurements in PD patients after implantation of STN electrodes. PMID- 16703592 TI - Trends in distribution and prognostic significance of Gleason grades on radical retropubic prostatectomy specimens between 1989 and 2001. AB - BACKGROUND: The objectives of the current study were to examine time trends in the prevalence of Gleason grades of prostate cancer on radical retropubic prostatectomy (RRP) specimens and to assess the resultant impact on prognosis. METHODS: The authors examined the prevalence over time of each grade and Gleason score (GS) on RRP specimens from 8750 patients who were treated between 1989 and 2001. Biochemical recurrence-free survival (BRFS), which was estimated by using Kaplan-Meier methodology, was examined in subgroups of patients defined by tumor grade and era of surgery. RESULTS: The prevalence of Grade 3 prostate cancers increased (86% vs. 49% for primary Gleason grade and 71% vs. 47% for secondary Gleason grade; 1999-2001 vs. 1989-1990, respectively), whereas the prevalence of Grade 2 tumors decreased (0.4% vs. 38% for primary Gleason grade and 1.3% vs. 28% for secondary Gleason grade, respectively) over the study period, leading to fewer GS 4 and 5 tumors and more GS 6 and 7 tumors. BRFS improved over time for patients who had GS 5 tumors (hazards ratio [HR], 0.92 per year; P = .003) and GS 6 tumors (HR, 0.93; P < .001) but remained unchanged for GS 7 tumors (HR 0.99; P = .462) and GS 8-10 tumors (HR 1.02; P = .360). Patients who were treated in the recent era (1997-2001) had greater differentiation of BRFS based on GS or Gleason grade compared with patients who were treated earlier (1989-1991). CONCLUSIONS: The current results confirmed that there were changes in the prevalence of Gleason grades on RRP specimens between 1989 and 2001. A chronological change in pathologic grading classification is suggested by evolving prognostic implications, which must be accounted for when comparing outcomes from different eras. PMID- 16703593 TI - Microanatomy and ultrastructure of the kidney of the African lungfish Protopterus dolloi. AB - The Dipnoi (lungfishes) have developed true lungs, having the ability to take oxygen from both the gills and the lungs. During the tropical dry season, many lungfish estivate on land, breathing only air. The estivation period is accompanied by profound functional modifications, including the suppression of urine. Thus, the lungfish kidney must be designed to cope with these dramatic cyclic changes in renal function. We study here the microanatomy and the structure of the kidney of the African lungfish Protopterus dolloi, maintained under controlled freshwater conditions. Chemical microdissection, light microscopy, and scanning and transmission electron microscopy have been used. The nephrons of P. dolloi are composed of a renal corpuscle (RC) and of a renal tubule that appears divided into five morphologically distinct segments: neck segment (NS), proximal tubule (PT), intermediate segment (IS), distal tubule (DT), and collecting tubule (CT). Paired CTs abut into a collecting duct, the latter emptying into an archinephric duct. The RCs lie in the mid-zone of the kidney, between the PTs and the convoluted DTs. The spatial distribution of these elements allows recognition of a kidney zonation. The RCs group into clusters (3 4 RCs per cluster) that are supplied by a single arteriole surrounded by pericytes. Each cluster appears to represent a functional unit with a common hemodynamic regulatory mechanism. The major processes of the podocytes form flattened networks that appear to constitute an integrated system due to the presence of gap junctions. The existence of mesangial cells with large cell processes, and of mesangial cells with a dendritic appearance, suggests a complex functional role (contractile and phagocytic) for the mesangium. The NS and the IS are the narrowest nephron segments, formed only by multiciliated cells. The PT and the DT can be subdivided, based on the tubular morphology and on cell composition, into portions I and II: PTI is formed only by brush border (BB) cells, while PTII contains BB and multiciliated cells. The DTI is formed by segment-specific cells, while the DTII contains segment-specific and a small number of flask cells. The CT contains principal and flask cells in a 5:1 ratio. The flask cells adopt two different configurations (with a narrow canaliculus or with a large cavity). The main goal of this study was to disclose specific kidney features that could be related to function, phylogeny, and habitat. In addition, the present results constitute the basis for a study of the morphologic changes that should occur in the kidney of P. dolloi during estivation. PMID- 16703594 TI - Fertility preservation for young people treated with cancer: what are the attitudes and knowledge of clinicians? PMID- 16703595 TI - The impact of tumor progesterone receptor status on optimal adjuvant endocrine therapy for postmenopausal patients with early-stage breast cancer: a decision analysis. AB - BACKGROUND: Emerging data suggest that treatment outcomes with aromatase inhibitors (AIs) and/or tamoxifen may differ for tumors that express both the estrogen receptor (ER) and the progesterone receptor (PR) (ER+/PR+) compared with those that lack PR expression (ER+/PR-). However, the optimal sequencing of AIs and tamoxifen as adjuvant therapy is not known and may differ for biologic subsets of cancers. METHODS: Markov models were used to simulate disease-free survival (DFS) separately among postmenopausal women with ER+/PR+ cancers and women with ER+/PR- cancers. By using risk estimates reported from randomized clinical trials, treatment with 5 years of an AI alone with sequential treatment consisting of tamoxifen with crossover to an AI at 2 years was compared. RESULTS: For women with ER+/PR+ cancers, sequential therapy with tamoxifen followed by crossover to an AI at 2 years yielded modest improvements in 10-year DFS estimates compared with planned AI monotherapy (84.3% vs. 82.2% and 68.8% vs. 64.8% for lymph node-negative and lymph node-positive patients, respectively). However, for women with ER+/PR- cancers, upfront treatment with an AI yielded improved outcomes with 10-year DFS rates of 90.5% and 80.1% for the lymph node negative and node-positive groups, respectively, compared with 88.2% and 76.1%, respectively, for sequential treatment with tamoxifen followed by an AI. CONCLUSIONS: Modeling estimates suggested that the optimal endocrine treatment strategy may differ based on the biologic features of breast cancer tumors. Patients with ER+/PR+ tumors achieved optimal 10-year DFS estimates with tamoxifen followed by a crossover to AI therapy, whereas patients with ER+/PR- tumors fared best when they initiated treatment with AI. PMID- 16703596 TI - Evaluation of glutathione S-transferase polymorphisms and mutagen sensitivity as risk factors for the development of second primary tumors in patients previously diagnosed with early-stage head and neck cancer. AB - BACKGROUND: The objective of this study was to evaluate the effects of polymorphisms in 2 genes in the glutathione S-transferase (GST) family and the mutagen-sensitivity phenotype on the risk of second primary tumors (SPTs) in patients with previously diagnosed early-stage head and neck squamous cell carcinoma. Data were available for 303 patients who were enrolled in a placebo controlled chemoprevention trial of low-dose 13-cis-retinoic acid to reduce the occurrence of SPTs. METHODS: A Cox proportional hazards model and survival tree analysis were used to evaluate the association between specified genetic variations and the development of SPTs. The average number of bleomycin-induced chromatid breaks per cell was used to quantify mutagen sensitivity as an individual patient's degree of sensitivity to genotoxicity. RESULTS: The GST-M1 null genotype was associated with an increased risk for any SPTs (hazard ratio [HR], 1.99; 95% confidence interval [95% CI], 1.11-3.56) and for tobacco-related SPTs (HR, 2.16; 95% CI, 1.01-4.62) after adjusting for covariates. The GST-T1 null genotype and bleomycin-induced chromatid breaks were not associated with a statistically significant increased risk for SPTs or tobacco-related SPTs after similar adjustment. Simultaneous nonnull status for both GST genotypes was associated with a decreased risk for any SPTs (HR, 0.52; 95% CI, 0.28-0.96) and tobacco-related SPTs (HR, 0.50; 95% CI, 0.22-1.11) compared with null status for GST-M1 accompanied by nonnull status for GST-T1. CONCLUSIONS: An association was observed between the development of SPTs and the GST-M1 null genotype after successful treatment for early-stage head and neck squamous cell carcinoma. The GST-T1 null genotype and bleomycin-induced chromatid breaks were not associated with an increased risk, and no significant interactions were identified. PMID- 16703597 TI - Allogeneic hematopoietic stem cell transplantation as part of postremission therapy improves survival for adult patients with high-risk acute lymphoblastic leukemia: a metaanalysis. AB - BACKGROUND: The prognosis for adult patients with acute lymphoblastic leukemia (ALL) remains unsatisfactory primarily because of the high incidence of recurrence. Therefore, optimal postremission therapy is a matter of vital concern. In particular, the clinical efficacy of allogeneic hematopoietic stem cell transplantation (HSCT) should be clarified. METHODS: Rigorous criteria were used to select 7 studies of adult ALL that prospectively assessed overall survival (OS) using natural randomization based on donor availability combined with intention-to-treat analyses. The authors then performed a metaanalysis to evaluate the role of allogeneic HSCT. RESULTS: Seven studies that included 1274 patients were selected. A metaanalysis demonstrated that patients in the donor groups had significantly better survival than patients in the no-donor groups (hazard ratio [HR], 1.29; 95% confidence interval [95% CI], 1.02-1.63 [P = .037]). When only high-risk patients were included in the analysis, the superiority of the survival advantage was even greater (HR, 1.42; 95% CI, 1.06 1.90 [P = .019]). A meta-regression analysis revealed that compliance with allogeneic HSCT showed a significant and positive correlation with survival (coefficient, 0.022; P < .01), suggesting that the greater the proportion of patients who actually received allogeneic HSCT, the better the survival of the donor group. No beneficial effects of autologous HSCT were observed. CONCLUSIONS: The current findings demonstrated that allogeneic HSCT improves the outcome of adult patients with high-risk ALL. Although these analyses were based on abstracted data, the results indicated that allogeneic HSCT should be considered for such patients if a suitable donor is available. PMID- 16703598 TI - The influence of age and comorbidity on receiving radiotherapy as part of primary treatment for cancer in South Netherlands, 1995 to 2002. AB - BACKGROUND: The objective of this study was to study the influence of age and comorbidity on receiving radiotherapy (RT) in primary treatment of cancer. METHODS: In a population-based setting, the authors calculated the proportion of irradiated patients within 6 months after they received a diagnosis of lung, rectal, breast, or prostate cancer or non-Hodgkin lymphoma (n = 33,369 patients) according to age and comorbidity between 1995 and 2002. Logistic regression analysis was used to adjust for age, comorbidity, gender, and stage. RESULTS: Patients with localized nonsmall cell lung cancer (NSCLC) ages 65 years to > or = 80 years or with comorbid conditions received RT alone significantly more often compared with younger patients (ages 65-79 years: odds ratio [OR], 3.4; age > or = 80: OR, 12.0) and patients without comorbidities (1 comorbid condition: OR, 2.1; > or = 2 comorbid conditions: OR, 2.4). This also applied to patients with nonlocalized NSCLC ages 65 years to 79 years compared with younger patients (OR, 1.4). RT was administered significantly less often to elderly patients with resected rectal cancers (ages 65-79 years: OR, 0.7; age > or = 80 years: OR, 0.4), patients age > or = 80 years with breast cancer after undergoing conserving surgery (OR, 0.1), and patients age > or = 80 years with clinical T1-T3,N0,M0 prostate cancer age (OR, 0.1) compared with younger patients. Patients with breast cancer who underwent breast-conserving surgery received RT significantly less often in the presence of comorbidities (1 comorbid condition: OR, 0.6; > or = 2 comorbid conditions: OR, 0.4). Older patients with aggressive non-Hodgkin lymphoma received only RT as treatment significantly more often compared with younger patients (OR, 3.4). CONCLUSIONS: Comorbidity and age did have influence over whether patients received RT, although, for most tumor types, age appeared to be a stronger predicting factor. Under treatment was observed among patients with breast cancer and rectal cancer. PMID- 16703600 TI - Complication of mediastinal mass: acquired tracheoesophageal fistula associated with T-cell lymphoblastic lymphoma. AB - The occurrence of a tracheoesophageal fistula (TEF) in the setting of lymphoma has only rarely been reported in the world literature. Most cases reported were associated with radiation therapy vs. chemotherapy alone. This report presents one case illustrating the difficulty encountered managing a TEF that developed while undergoing chemotherapy for T-cell lymphoblastic lymphoma. PMID- 16703599 TI - Effects of the dual 5 alpha-reductase inhibitor dutasteride on apoptosis in primary cultures of prostate cancer epithelial cells and cell lines. AB - BACKGROUND: The profound reduction in serum dihydrotestosterone (DHT) observed with the dual 5 alpha-reductase inhibitor (5ARI) dutasteride makes it an attractive agent for prostate cancer therapy. The objective of the current study was to determine whether dutasteride would induce apoptosis in a range of prostate epithelial cell lines and primary cultures. METHODS: Both human prostate androgen-sensitive cell lines (PwR-1E, PNT-2, LNCaP, and PC3[AR2]) and an androgen-independent cell line (PC-3) were grown to confluence. Primary epithelial cells extracted from fresh prostate cancer radical prostatectomy specimens also were grown to confluence under optimal conditions. Total cellular protein was extracted to confirm cytokeratin 18 and antihuman alpha-methylacyl CoA racemase (AMACR) expression of the primary cells. Apoptosis was assessed by propidium iodide DNA staining and flow cytometry after 24 hours of culture in from 0 microM to 10 microM of dutasteride. RESULTS: Dutasteride induced a dose dependent increase in apoptosis in the androgen-sensitive prostate cell lines PwR 1E, PNT-2, and LNCaP and in the androgen receptor-expressing PC3(AR2) cell line. However, there was no significant apoptosis noted in the parental PC-3 cells. Of 16 primary epithelial cultures that were treated, 7 cultures were induced to undergo apoptosis, and 9 cultures were unresponsive. All primary cultures were positive for cytokeratin 18 expression, confirming their epithelial phenotype. Responder epithelial cells were positive for AMACR expression. CONCLUSIONS: The results of the current study confirmed that dutasteride differentially induced apoptosis in a subset of prostate cell lines and primary prostate epithelial cells. Understanding the cellular phenotype may indicate susceptible cells. PMID- 16703601 TI - An overview of posttraumatic stress disorder in African Americans. AB - While several studies have found high rates of trauma exposure there is limited information on posttraumatic stress disorder (PTSD) and its relationship to depression in the African American population. The prevalence and/or expression of psychiatric disorders can differ between racial/ethnic groups. The authors review literature addressing trauma exposure, prevalence, and expression of PTSD in the African American population. Risk factors that may be of specific significance to the development of PTSD in African Americans are also reviewed. Additionally, treatment issues and potential directions for future research of PTSD in the African American population are discussed. PMID- 16703602 TI - Impact of PTSD comorbidity on one-year outcomes in a depression trial. AB - Low-income African American, Latino, and White women were screened and recruited for a depression treatment trial in social service and family planning settings. Those meeting full criteria for major depression (MDD; N = 267) were randomized to cognitive-behavior therapy (CBT), antidepressant medication, or community mental health referral. All randomly assigned participants were evaluated by baseline telephone and clinical interview, and followed by telephone for one year. Posttraumatic stress disorder (PTSD) comorbidity was assessed at baseline and one-year follow-up in a clinical interview. At baseline, 33% of the depressed women had current comorbid PTSD. These participants had more exposure to assaultive violence, had higher levels of depression and anxiety, and were more functionally impaired than women with depression alone. Depression in both groups improved over the course of one year, but the PTSD subgroup remained more impaired throughout the one-year follow-up period. Thus, evidence-based treatments (antidepressant medication or structured psychotherapy) decrease depression regardless of PTSD comorbidity, but women with PTSD were more distressed and impaired throughout. Including direct treatment of PTSD associated with interpersonal violence may be more effective in alleviating depression in those with both diagnoses. PMID- 16703603 TI - The roles of religion and spirituality among African American survivors of domestic violence. AB - The investigation examined religious involvement, spirituality, religious coping, and social support as correlates of posttraumatic stress symptoms and depression symptoms in African American survivors of domestic violence. Sixty-five African American women who experienced domestic violence in the past year provided data on demographics, severity and frequency of physical and psychological abuse during the past year, aspects of current social support, types of current coping activities, religious involvement, spiritual experiences, and symptoms related to depression and posttraumatic stress disorder. Women who evinced higher levels of spirituality and greater religious involvement reported fewer depression symptoms. Religious involvement was also found to be negatively associated with posttraumatic stress symptoms. Women who reported higher levels of spirituality reported utilizing higher levels of religious coping strategies, and women who reported higher levels of religious involvement reported higher levels of social support. Results did not support hypotheses regarding social support and religious coping as mediators of the associations between mental health variables, religious involvement, and spirituality. PMID- 16703604 TI - Ethnography's role in assisting mental health research and clinical practice. AB - In this article, the author discusses the utility of qualitative approaches in mental health research and clinical practice. He highlights the value of conceptualizing research designs and procedures that include relevant cultural information about study participants. Emphasis is placed on African American women participants, consumers, and others who are understudied, underrepresented, and underserved in large-scale studies and treatment. Limitations and strengths are noted. Contributions of qualitative methodology are described, particularly the functions of ethnography and grounded theory in conceptualization, hypotheses building, and hypotheses testing of outcome data on African American women and culturally diverse groups. Clinical researchers are encouraged to utilize qualitative and mixed methods approaches to bolster outcomes and better inform their clinical practice. PMID- 16703605 TI - Clinical depression in African American women: diagnoses, treatment, and research. AB - Although there is some debate about the relative prevalence rates of depression in African Americans compared to Whites, there is little debate among investigators that there is a shortage of research data on depression in African Americans generally, and more specifically on African American women. Lack of adequate and sufficient research on African Americans contributes to the problems of misdiagnoses, under-diagnoses, and undertreatment of depression in African Americans, generally, and more specifically, African American women, the focus of this special section. African American women are understudied, underserved, and misdiagnosed as a group. A paucity of studies on depression in African American women has existed for decades. This special series of papers will highlight the existing problem of depression in African American women with a focus on diagnostic, treatment, and research issues. PMID- 16703606 TI - The use of psychiatric medications to treat depressive disorders in African American women. AB - Review of the current literature confirms that African American women as a group are underdiagnosed and undertreated for psychiatric disorders. Hence, much effort is targeted towards awareness, screening, and improving access to health care for this population. However, once an African American woman is diagnosed with a major mental health disorder, determining the optimal course of treatment is a process that must be approached carefully because of gender and racial/ethnic differences in response and metabolism of psychiatric medications. African American women fall into both of these understudied categories. Given the small numbers of African American women represented in the clinical trials on which clinical practice is based, one must consider the limitations of current knowledge regarding psychoactive medications in this population. Culturally based attitudes or resistance to pharmacotherapy can complicate the use of psychoactive medicines, often a first-line approach in primary care clinics. Communication with patients is key, as well as openness to patient concerns and tolerance of these medications. PMID- 16703608 TI - The influence of freeze duration on postfreeze recovery by caterpillars of Pyrrharctia isabella (Lepidoptera: Arctiidae): when is survival enough to qualify as recovery? AB - The effects of freeze exposure at -4.5 degrees C for as long as 6 weeks on long term postfreeze survival and body fluid composition were investigated in caterpillars (woolly bears) of the arctiid moth Pyrrharctia isabella. Woolly bears routinely survived the initial postfreeze period with little difference between the 1-week (100%) and 6-week (95%) freeze treatments. Caterpillars in the latter treatment, however, reached the pupal stage almost one half as often as woolly bears in the 1-week freeze treatment. The success rate for adult emergence was not different for pupae from the two treatments (ca. 50%). Woolly bears responded to cold acclimation by accumulating glycerol to levels exceeding 300 mM although this was not augmented by extending the acclimation period to 6 weeks. There was a significant (P<0.05) rise in hemolymph [K+] during the first week of the freeze (23.4-37.8 mM), which then remained stable over the remainder of the 6 week freeze period. Hemolymph [Na+] did not change from the prefreeze level over the course of the freeze treatment. Body water content showed a modest rise during the course of the freeze treatment but the underlying cause for this change was uncertain. Prolonged freeze exposure had a major impact on long-term survival of the woolly bears but this was not reflected by any instability in body fluid composition. Moreover, short-term recovery was not an effective indicator of the tolerance of P. isabella caterpillars to prolonged freezing. PMID- 16703607 TI - Chromatic effects of endothelin family peptides in non-innervated fish, Synbranchus marmoratus, melanophores. AB - The biological activity of endothelins (ETs) in non-innervated Synbranchus marmoratus melanophores was demonstrated. These peptides induced a dose-dependent pigment aggregation (lightening skin) in these cells. However, they presented EC50's (effective concentration required to produce 50% of response) 26, 106 and 35 times higher than, respectively, the melanin concentrating hormone (MCH) EC50, and exhibited a characteristic temporal and dose-dependent autodessensibilization of the aggregative effect on the melanophores of this fish. The receptor characterization suggested the presence of the ET(B) subtype, since BQ-788 (selective antagonist of ET(B)) but not BQ-485 (selective antagonist of ET(A)) blocked the aggregative effect of the hormones. Confirming these data, sarafotoxin (SRTX) S6c, a toxin selective for ET(B), induced maximal aggregation of pigment granules. S6c presented an EC50 6.8 times higher than the MCH EC50, and 3.9, 15.6 and 5.1 times lower than the EC50's ETs, respectively. The melanotropic effect of SRTX S6b and vasoactive intestinal contractor (VIC) were demonstrated for the first time in this work. SRTX S6b induced a dose-dependent pigment aggregation and presented an EC50 2.54 and 17.2 times higher than the S6c and MCH EC50's, respectively. Compared to the ETs it was 1.53, 6.19 and 2.03 times lower, respectively. PMID- 16703609 TI - "Evolutionist and missionary," The Reverend John Thomas Gulick (1832-1923). Part I: cumulative segregation--geographical isolation. AB - During the late 19th and early 20th centuries, the American missionary and naturalist John Thomas Gulick (1832-1923) was one of the most well-known and influential evolutionists, anticipating in his research and writing, later proposals of geographical isolation, population genetics, genetic drift, and the founder principle in speciation. In over 20 publications based on studies of non adaptive geographical variation in several hundred species of snails in the genus Achatinella, collected in the valleys of Oahu in the Sandwich Islands (Hawaii), Gulick provided evidence for the formation of new species from varieties and the importance of geographical (non-adaptive) isolation in species diversification. Gulick's theory of the species-differentiating effects of isolation was regarded by many as a more complete theory of speciation than Darwin's, and by others as correcting a fundamental deficiency in Darwin's theory, namely how groups of organisms diversify one from another. Gulick also saw organisms as active participants in, and in interaction with, their environment, for which he coined the term coincident selection, anticipating the Baldwin effect/organic selection. With his concepts of cumulative segregation (geographical isolation), indiscriminate isolation (the Founder effect) and coincident selection (the Baldwin effect), we should recognize Gulick as one of the earliest and most original and innovative evolutionary biologists. PMID- 16703610 TI - Three-dimensional structure of the zona pellucida at ovulation. AB - The mammalian zona pellucida (ZP) is an extracellular matrix surrounding oocytes and early embryos, which is critical for normal fertilization and preimplantation development. It is made up of three/four glycoproteins arranged in a delicate filamentous matrix. Scanning electron microscopy (SEM) studies have shown that ZP has a porous, net-like structure and/or nearly smooth and compact aspect. In this study, the fine 3-D structure of the human and mouse ZP is reviewed with the aim to integrate ultrastructural and molecular data, considering that the mouse is still used as a good model for human fertilization. By conventional SEM observations, numerous evidences support that the spongy ZP appearance well correlates with mature oocytes. When observed through more sophisticated techniques at high resolution SEM, ZP showed a delicate meshwork of thin interconnected filaments, in a regular alternating pattern of wide and tight meshes. In mature oocytes, the wide meshes correspond to "pores" of the "spongy" ZP, whereas the tight meshes correspond to the compact parts of the ZP surrounding the pores. In conclusion, the traditional "spongy" or "compact" appearance of the ZP at conventional SEM appears to be only the consequence of a prevalence of different arrangements of microfilament networks, according to the maturation stage of the oocyte, and in agreement with the modern supramolecular model of the ZP at the basis of egg-sperm recognition. Despite great differences in molecular characterization of ZP glycoproteins between human and mouse ZP, there are no differences in the 3-D organization of glycoproteic microfilaments in these species. PMID- 16703611 TI - Structural bases of the ovarian function: an introduction. PMID- 16703612 TI - Presence and significance of temperature gradients among different ovarian tissues. AB - After recalling male gonadal physiology in respect of tissue temperatures within the scrotal sac, and raising questions concerning abdominal testes, attention turned to mature Graafian follicles and ovarian stroma. Temperature gradients between such tissues were summarized for human, rabbit, pig, and cow, and generally fell in the range of 1.3-1.7 degrees C: follicles were always cooler than stroma. Measurements were made principally by means of a thermo-sensing camera at midventral laparotomy, but also using microelectrodes or thermistor probes sited in the follicular antrum of rabbits and pigs, respectively. When thermo-imaged under the fimbriated extremity of the Fallopian tube, mature pig follicles and stroma could still be distinguished. Such follicles cooled slightly more rapidly during the first 10 s of a 60-s recording interval, after which curves for the two tissues remained parallel. Arresting ovarian blood supply for 5 min had a negligible influence on the temperature differentials. Endoscopy in three models recorded mean differentials of 0.6 +/- 0.1 degrees C - 1.1 +/- 0.1 degrees C between follicles and stroma, but such follicles had not attained mature diameter. Temperature gradients were thought to be generated at least in part by endothermic reactions within mature follicles, reflecting hydration of large extracellular matrix molecules such as proteoglycans. A contribution to the cooling process from the products of leukocyte activity in the follicle wall and antrum could also be involved. Temperature gradients would be maintained locally by counter-current heat exchange mechanisms and, in this context, the microvasculature and lymphatic flow of individual follicles were found to be appropriate. Observations on the temperature of preovulatory follicles appear relevant to procedures of in vitro maturation and in vitro fertilization. PMID- 16703613 TI - Immune system involvement in the regulation of ovarian function and augmentation of cancer. AB - Increasing evidence indicates a role for the immune system and mesenchymal epithelial interactions in the regulation of ovarian function. Cytokines produced by mesenchymal cells can stimulate development and regression of ovarian structures. We report here that mesenchymal cells releasing surface molecules among epithelial cells--namely vascular pericytes and monocyte-derived cells (MDC)--and intraepithelial T lymphocytes are associated with oogenesis and formation of new primary follicles in both fetal and adult human ovaries. These activated mesenchymal cells interact with the ovarian surface epithelium, which appears to be a source of secondary germ cells and granulosa cells. Activated pericytes and MDC are also associated with stimulation of thecal development during selection of growing secondary follicles from the cohort of primary follicles. However, survival of the dominant follicle during mid-follicular phase selection is associated with a lack of activity of mesenchymal cells and retardation of thecal development, since immature granulosa cells lacking aromatase are unable to resist high levels of thecal androgens. Once the selected follicle matures (late follicular phase), it shows enhanced activity of thecal mesenchymal cells and advanced thecal development. Corpus luteum (CL) development is accompanied by a high activity of vascular pericytes and MDC. In mature CL and CL of pregnancy, luteal MDC and pericytes show a stable (inactive) state. Regression of the CL is associated with regression of pericytes, transformation of MDC into dendritic cells, infiltration by T lymphocytes, and binding of immunoglobulin G to the luteal cells. The immunoglobulin M (IgM) binds to young but not mature luteal cells. In the CL of pregnancy, IgM binds to luteal vessels, but not to luteal cells. Regressing CL shows IgM binding to both luteal cells and vessels. In ovarian cancers, highly activated MDC and sometimes activated pericytes (poorly differentiated carcinomas) are present. IgM binding is similar to that seen in the CL of pregnancy. These data indicate that vascular pericytes, MDC, T cells, and immunoglobulins may play an important role in the regulation of ovarian physiology and contribute to the augmentation of ovarian cancer growth. PMID- 16703615 TI - Effect of corazonin and crustacean cardioactive peptide on heartbeat in the adult American cockroach (Periplaneta americana). AB - Changes in the frequency of cardiac pulsations have been monitored in the decapitated body of adult P. americana before and 5 h after the injections of [Arg(7)]-corazonin and CCAP, using newly invented touch-free, noninvasive optocardiographic methods. Relatively large dosages of these peptides (10(-6) M concentrations in the body) had no effect on the rate of the heartbeat beyond the Ringer control limits. It has been concluded, therefore, that Corazonin and CCAP, which are currently cited in the literature as "the most potent cardiostimulating peptides" in insects, have no effect on the physiological regulation of cardiac functions in the living body. PMID- 16703614 TI - Cloning and nucleotide sequencing of three heat shock protein genes (hsp90, hsc70, and hsp19.5) from the diamondback moth, Plutella xylostella (L.) and their expression in relation to developmental stage and temperature. AB - Heat shock protein genes, hsp90, hsc70, and hsp19.5, were cloned and sequenced from the diamondback moth, Plutella xylostella (L.) by RT-PCR and RACE method. The cDNA sequence analysis of hsp90 and hsp19.5 revealed open reading frames (ORFs) of 2,151 and 522 bp in length, which encode proteins with calculated molecular weights of 82.4 and 19.5 kDa, respectively. Analysis of cDNA from hsc70 revealed an ORF of 1,878 bp coding a protein with a calculated molecular weight of 69.3 kDa. Furthermore, the analysis of genomic DNA from hsc70 confirmed the presence of introns while no introns were apparent in hsp90 and hsp19.5. Southern blot analysis suggested the presence of multiple copies of each gene family in the DBM genome. Detectable expression of hsp19.5 was observed at the pupal stage while expression of hsp90 and hsc70 was detected at both pupal and adult stages. At adult stage, females showed a higher expression of hsp90 and hsc70 than males. An increased expression was observed in all three genes after exposure to a high temperature in both sexes. These results suggest that in addition to a heat shock response, these HSP genes might be involved in other functions during the course of development in DBM. PMID- 16703616 TI - Silencing allatostatin expression using double-stranded RNA targeted to preproallatostatin mRNA in the German cockroach. AB - YXFGL-NH(2) family allatostatins (ASTs) were isolated from cockroach brain extracts based on their capacity to inhibit juvenile hormone (JH) biosynthesis in corpora allata (CA) incubated in vitro. Subsequently, the inhibitory activity of synthetic ASTs was demonstrated experimentally, although these peptides were shown to be active as JH inhibitors only in cockroaches, crickets, and termites. Here, we sought to examine whether ASTs are true physiological regulators of JH synthesis. To this end, we used RNA interference methodologies and the cockroach Blattella germanica as a model. Treatments with double-stranded RNA targeting the allatostatin gene in females of B. germanica produced a rapid and long-lasting reduction in mRNA and peptide levels in both brain and midgut during the reproductive cycle. Nevertheless, while brain AST levels were reduced approximately 70-80%, JH synthesis did not increase in any of the age groups tested. PMID- 16703618 TI - Normal embryonic development and cardiac morphogenesis in mice with Wnt1-Cre mediated deletion of connexin43. AB - Mice harboring a null mutation in the gap junction protein connexin43 (Cx43) die shortly after birth due to an obstruction of the right ventricular outflow tract of the heart. These hearts exhibit prominent pouches at the base of the pulmonary outlet, i.e., morphological abnormalities that were ascribed to Cx43-deficiency in neural crest cells. In order to examine the Cx43 expression pattern in neural crest cells and derived tissues and to test whether neural crest-specific deletion of Cx43 leads to the conotruncal defects seen in Cx43null mice, we ablated Cx43 using a Wnt1-Cre transgene. Deletion of Cx43 was complete and occurred in neural crest cells as well as in neural crest-derived tissues. Nevertheless, hearts of mice lacking Cx43 specifically in neural crest cells were indistinguishable from controls. Thus, the morphological heart abnormalities of Cx43 null mice are most likely not caused by lack of Cx43 in neural crest cells. PMID- 16703617 TI - Glucose transporter 8 (GLUT8) from the red imported fire ant, Solenopsis invicta Buren (Hymenoptera: Formicidae). AB - We have cloned the fire ant glucose transporter 8 (GLUT8) cDNA providing the first molecular characterization of a GLUT8 in insects. Glucose is a poly-alcohol and, due to its high hydrophilicity, cannot move across cell membranes. GLUT8 is a putative facilitative transporter for the cellular import and export of glucose. The complete 2,974-bp cDNA encodes a 501-residue protein with a predicted molecular mass of 54.8 kDa. Transcripts were detected in the brain, midgut, hindgut, Malpighian tubule, fat body, ovary, and testis. The highest transcriptional expression was found in fat body. Northern blot analysis revealed different transcript sizes in mated queen brains, alate female ovaries, and male testes. We propose that four other sequences obtained from insect genome projects from the honey bee Apis mellifera (ENSAPMP00000006624), the malaria mosquito Anopheles gambiae (EAA11842), and the fruit fly Drosophila melanogaster (AAQ23604 and AAM52591) are likely the orthologues of the fire ant GLUT8. Phylogenetic relationships in insect glucose transporters are presented. PMID- 16703619 TI - Experimental analysis of lens-forming capacity in Xenopus borealis larvae. AB - Previously, the only anuran amphibians known to have the capacity to regenerate a lens after lentectomy were Xenopus laevis and Xenopus tropicalis. This regeneration process occurs during the larval life through transdifferentiation of the outer cornea promoted by inductive factors produced by the retina and accumulated inside the vitreous chamber. However, the capacity of X. tropicalis to regenerate a lens is much lower than that of X. laevis. This study demonstrates that Xenopus borealis, a species more closely related to X. laevis than to X. tropicalis, is not able to regenerate a lens after lentectomy. Nevertheless, some morphological modifications corresponding to the first stages of lens regeneration in X. laevis were observed in the outer cornea of X. borealis. This suggested that in X borealis the regeneration process was blocked at early stages. Results from histological analysis of X. borealis and X. laevis lentectomized eyes and from implantation of outer cornea fragments into the vitreous and anterior chambers demonstrated that: (i) in X. borealis eye, the lens-forming competence in the outer cornea and inductive factors in the vitreous chamber are both present, (ii) no inhibiting factors are present in the anterior chamber, the environment where lens regeneration begins, (iii) the inability of X. borealis to regenerate a lens after lentectomy is due to an inhibiting action exerted by the inner cornea on the spreading of the retinal factor from the vitreous chamber towards the outer cornea. This mechanical inhibition is assured by two distinctive features of X. borealis eye in comparison with X. laevis eye: (i) a weaker and slower response to the retinal inducer by the outer cornea; (ii) a stronger and faster healing of the inner cornea. Unlike X. tropicalis and similar to X. laevis, in X. borealis the competence to respond to the retinal factor is not restricted to the corneal epithelium but also extends to the pericorneal epidermis. PMID- 16703620 TI - Competition between Nitrospira spp. and Nitrobacter spp. in nitrite-oxidizing bioreactors. AB - In this work the question was addressed if in nitrite-oxidizing activated sludge systems the environmental competition between Nitrobacter spp. and Nitrospira spp., which only recently has been discovered to play a role in these systems, is affected by the nitrite concentrations. Two parallel chemostats were inoculated with nitrifying-activated sludge containing Nitrospira and operated under identical conditions. After addition of Nitrobacter to both chemostats, the nitrite concentration in the influent of one of the chemostats was increased such that nitrite peaks in the bulk liquid of this reactor were detected. The other chemostat served as control reactor, which always had a constant nitrite influent concentration. The relative cellular area (RCA) of Nitrospira and Nitrobacter was determined by quantitative fluorescence in situ hybridization (FISH). The nitrite perturbation stimulated the growth of Nitrobacter while in the undisturbed control chemostat Nitrospira dominated. Overall, the results of this experimental study support the hypothesis that Nitrobacter is a superior competitor when resources are abundant, while Nitrospira thrive under conditions of resource scarcity. Interestingly, the dominance of Nitrobacter over Nitrospira, caused by the elevated nitrite concentrations, could not be reverted by lowering the available nitrite concentration to the original level. One possible explanation for this result is that when Nitrobacter is present at a certain cell density it is able to inhibit the growth of Nitrospira. An alternative explanation would be that the length of the experimental period was not long enough to observe an increase of the Nitrospira population. PMID- 16703621 TI - Systematic review of outcome of synchronous portal-superior mesenteric vein resection during pancreatectomy for cancer. AB - BACKGROUND: Tumour clearance during pancreatectomy may be facilitated by resection of the portal-superior mesenteric vein, but this is associated with increased perioperative risk. There is no consensus about which patients benefit from portal-superior mesenteric vein resection. METHODS: A systematic appraisal was carried out of the literature on portal-superior mesenteric vein resection during pancreatectomy to identify recurrent themes to guide management. A computerized search of the Medline and Embase databases found 52 non-duplicated studies providing relevant data in 1646 patients. Pooled data were examined for information on outcome categories relating to operation, complications, histopathology and overall outcome. RESULTS: The median (range) number of patients with portal-superior mesenteric vein resection per cohort was 23 (4 172). Median operating time was 513 (168-1740) min and blood loss 1750 (300 26000) ml. Postoperative morbidity ranged from 9 to 78 per cent with a median per cohort of 42 per cent. There were 73 perioperative deaths (5.9 per cent of 1235 for whom mortality data were provided). Median survival was 13 months, and 1-, 3- and 5-year survival rates were 50, 16 and 7 per cent respectively. Specimen histopathology confirmed positive nodes in 67.4 per cent. CONCLUSIONS: This is the largest collective report to date on portal-superior mesenteric vein resection in pancreatectomy. The high rate of nodal metastases and low 5-year survival rates suggest that by the time of tumour involvement of the portal vein cure is unlikely, even with radical resection. PMID- 16703622 TI - Prevalence of glucose intolerance in primary hyperparathyroidism and the benefit of parathyroidectomy. AB - BACKGROUND: Increased prevalence of diabetes mellitus (DM) in primary hyperparathyroidism (PHPT) is established, but not glucose intolerance (GI), nor benefit from parathyroidectomy on GI. We determined these during management of a continuous series of patients with PHPT routinely followed after surgery. PATIENTS AND METHODS: WHO criteria classified 75 g oral glucose tolerance tests (OGTT) in 51/54 consecutively proven PHPT patients, into normal glucose tolerance (NGT), DM, impaired glucose tolerance (IGT) or impaired fasting glucose (IFG); GI was derived by adding those with DM and IGT/IFG. OGTT were repeated after parathyroidectomy (mean follow up 2.4 +/- SD 1.6 years). Paired student t tests were used to compare fasting and 2-h plasma glucose (PG). RESULTS: At presentation 32/54 patients (59%) had NGT, 10 IGT/IFG (19%) and 12 type 2 DM (22%), nine newly diagnosed. Before parathyroidectomy 17/35 patients had NGT (49%), 18 GI (51%), 12 DM (34%) and 6 IGT/IFG (17%). Five out of six patients with IGT/IFG had NGT, one with NGT developed IGT. At completion 23 patients (66%) had NGT, 12 GI (34%), 4 IGT/IFG (11%) and 8 DM (23%). After parathyroidectomy fasting and 2-h. PG fell in 30/34 normocalcaemic patients not on hypoglycaemic agents, 5.6 +/- 1.0 to 5.4 +/- 0.8 mmol/l, 7.2 +/- 3.0 to 6.3 +/- 3.1 mmol/l (p < 0.05, p < 0.01). CONCLUSIONS: 1. At presentation with PHPT, OGTT commonly identifies Type 2 DM and GI.2. After successful parathyroidectomy fasting and 2 h. PG fall significantly (p < 0.05, p < 0.01). DM and IGT/IFG often ameliorates to IGT or NGT, persistently. PMID- 16703623 TI - Defunctioning stomas in the treatment of rectal cancer. PMID- 16703624 TI - Randomized clinical trial to evaluate the effects of perioperative supplemental oxygen administration on the colorectal anastomosis. AB - BACKGROUND: Perioperative supplemental oxygen therapy may have beneficial effects on wound healing following colorectal surgery. The aim of this study was to evaluate the effects of such therapy on colorectal anastomotic pH and partial pressure of carbon dioxide (PCO(2)) gap. METHODS: Forty-five patients undergoing anterior resection for rectal or sigmoid cancer were randomized to receive 30 or 80 per cent perioperative oxygen. Administration was commenced after induction of anaesthesia and maintained for 6 h after surgery. Intragastric and anastomotic tonometric catheters were placed in each patient and intramucosal pH (pHi) was measured immediately after operation, and 6 and 24 h later. Gastric and anastomotic pHi and PCO(2) gap in each group were compared. RESULTS: There was a significantly lower anastomotic pHi and wider PCO(2) gap for gastric readings in the 30 per cent O(2) group, both 30 min (pHi, P = 0.006; PCO(2) gap, P = 0.006) and 6 h (pHi, P = 0.024; PCO(2) gap, P = 0.036) after surgery. There were no differences 24 h after surgery while breathing room air (pHi, P = 0.131; PCO(2) gap P = 0.139). No difference was found between gastric and anastomotic readings at any time point in the 80 per cent O(2) group. CONCLUSION: Perioperative administration of 80 per cent O(2) both during surgery and for 6 hours afterwards is associated with an improvement in relative anastomotic hypoperfusion as assessed by the measurement of pHi and PCO(2) gap. PMID- 16703625 TI - Stacking, derivatization, and separation by capillary electrophoresis of amino acids from cerebrospinal fluids. AB - This paper describes the in-column derivatization, stacking, and separation of amino acids by CE in conjunction with light-emitting diode-induced fluorescence using naphthalene-2,3-dicarboxaldehyde (NDA). According to the relative electrophoretic mobilities and the migration direction in tetraborate solution (pH 9.3), the injection order is cyanide, then amino acids, then NDA. Once poly(ethylene oxide) (PEO) migrates through the capillary under EOF, the amino acid.NDA derivatives, amino acids, and CN- ions migrating against the EOF enter the PEO zone. As a result of increases in viscosity and possible interactions with PEO molecules, the reagents/analytes slow down such that they become stacked at the boundary. In comparison with the off-column approach to the analysis of amino acids, our proposed method provides a lower degree of interference from polymeric NDA compounds and other side products. As a result, the plot of the peak height as a function of gamma-aminobutyric acid (GABA) concentration is linear over the range from 10(-5) to 10(-8) M, with the LOD being 4 nM. We demonstrate the diagnostic potential of this approach for the determination of amino acids, including GABA and glutamine, in biological samples through the analysis of large volumes of cerebral spinal fluids without the need for sample pretreatment. PMID- 16703626 TI - Measuring D-serine efflux from mouse cortical brain slices using online microdialysis-capillary electrophoresis. AB - Efflux of a number of important neurochemicals, including D-serine, L-serine, taurine, glutamate, and gamma-aminobutyric acid (GABA), from mouse cortical brain slices housed in a 7 microL perfusion chamber was monitored using online microdialysis-CE (MD-CE). Analyte concentrations could be measured every 20-27 s using the MD-CE instrument. Stimulation with high potassium induced increased release of D-serine. Kainic acid (KA) induced D-serine release, but this release was not blocked by 6-cyano-7-nitroquinoxaline-2,3-dione, suggesting that alpha amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid/KA receptors do not mediate D serine release. Application of L-serine, the precursor of D-serine, resulted in increased extracellular D-serine concentrations. L-Cysteine also increased extracellular D-serine levels in a partially Na+-dependent manner. The observed effects upon application of L-serine and L-cysteine support the involvement of ASC neutral amino acid transporters in regulating the extracellular concentration of D-serine concentration through competitive inhibition of uptake or increased release through heteroexchange. PMID- 16703627 TI - Analysis of protein-protein interactions with a multi-capillary electrophoresis instrument. AB - Protein-protein interactions were analyzed by zone electrophoresis of premixed equilibrium mixtures of a fluorescence-labeled protein at a constant concentration and unlabeled protein at a variety of concentrations using a 96-CE instrument equipped with a LIF detector. The interactions between labeled-con A versus succinylated ovalbumin, labeled-trypsin versus four proteinaceous trypsin inhibitors and labeled-insulin versus seven anti-insulin monoclonal antibodies were analyzed using a dual buffer system, in which a 60 mM borate-Na buffer (pH 9.35) was used as electrophoresis buffer and 60 mM MOPS-Na (pH 7.35) containing 0.1% Tween 20 was used as a sample buffer. The dual buffer system allowed fast and reproducible analyses of interactions at a physiological pH using uncoated fused-silica capillaries. The change in the mobility moment, the first statistical moment of an electropherogram on the mobility axis (Shimura, K., Uchiyama, N., Enomoto, M., Matsumoto, H., Kasai, K., Anal. Chem. 2005, 77, 564 572), of the labeled proteins were analyzed as a function of the concentration of unlabeled proteins. The dissociation constants for seven antibodies ranging from sub nanomolar to micromolar was determined based on the results of one cycle of parallel electrophoresis runs, which completed in 30 min using 20 pmol (120 ng) of labeled insulin and 5 pmol (750 ng) each of the mAb. PMID- 16703628 TI - Separation and detection of VX and its methylphosphonic acid degradation products on a microchip using indirect laser-induced fluorescence. AB - The application of indirect LIF (IDLIF) technique for on-chip electrophoretic separation and detection of the nerve agent O-ethyl S-[2-(diisopropylamino)ethyl] methylphosphonothiolate (VX) and its major phosphonic degradation products, ethyl methylphosphonic acid (EMPA) and methylphosphonic acid (MPA) was demonstrated. Separation and detection of MPA degradation products of VX and the nerve agent isopropyl methylphosphonofluoridate (GB) are presented. The negatively charged dye eosin was found to be a good fluorescent marker for both the negatively charged phosphonic acids and the positively charged VX, and was chosen as the IDLIF visualization fluorescent dye. Separation and detection of VX, EMPA, and MPA in a simple-cross microchip were completed within less than a minute, and consumed only a 50 pL sample volume. A characteristic system peak that appeared in all IDLIF electropherograms served as an internal standard that increased the reliability of peak identification. The negative peak of both VX and the MPAs is in agreement with indirect detection theory and with previous reports in the literature. The LOD of VX and EMPA by IDLIF was 30 and 37 microM, respectively. Despite the fact that the detection sensitivity is relatively low, the rapid simultaneous on-chip analysis of both VX and its degradation products as well as the separation and detection of the MPA degradation products of both VX and GB, increases detection reliability and may present a choice when sensitivity is not critical compared with speed and simplicity of the assay. PMID- 16703630 TI - 2-D native-PAGE/SDS-PAGE visualization of an oligomer's subunits: application to the analysis of IgG. AB - A 2-D native-PAGE/SDS-PAGE method for detecting the subunit components of protein oligomers at low picomole sensitivity is presented. IgG was electrophoresed in a native acidic polyacrylamide gel in amounts ranging from 51 pmol to 60 fmol. Silver-staining (native fast silver stain, ammoniacal silver stain, permanganate silver stain), Coomassie-staining (R-250, G-250), metal ion-reverse-staining (zinc, copper), and fluorescent chromophore-staining (SYPRO Ruby) methods were used to visualize the IgG oligomers. The protein zones were then excised, separated by SDS-PAGE, and subunits visualized with a permanganate silver stain. The Coomassie R-250/permanganate silver-staining combination detected IgG subunits using 2 pmol of sample. Coomassie G-250 and native fast silver staining in the first-dimensional gel produced detectable subunits in the second dimensional separation at 3 and 13 pmol, respectively. Staining with silver (ammoniacal, permanganate), copper, zinc, or SYPRO Ruby in the first-dimensional gel did not produce discernible subunits in the second-dimensional gels due to protein streaking or protein immobilization in the native gel. When using a 2-D native-PAGE/SDS-PAGE system, Coomassie staining of the first-dimensional native gel combined with permanganate silver staining of the second-dimensional denaturing gel provides the most sensitive method (2-3 pmol) for visualizing constituent subunits from their oligomeric assemblies. PMID- 16703632 TI - Study of Joule heating effects on temperature gradient in diverging microchannels for isoelectric focusing applications. AB - IEF is a high-resolution separation method taking place in a medium with continuous pH gradients, which can be set up by applying electrical field to the liquid in a diverging microchannel. The axial variation of the channel cross sectional area will induce nonuniform Joule heating and set up temperature gradient, which will generate pH gradient when proper medium is used. In order to operationally control the thermally generated pH gradients, fundamental understanding of heat transfer phenomena in microfluidic chips with diverging microchannels must be improved. In this paper, two 3-D numerical models are presented to study heat transfer in diverging microchannels, with static and moving liquid, respectively. Through simulation, the temperature distribution for the entire chip has been revealed, including both liquid and solid regions. The model for the static liquid scenario has been compared with published results for validation. Parametric studies have showed that the channel geometry has significant effects on the peak temperature location, and the electrical conductivity of the medium and the wall boundary convection have effects on the generated temperature gradients and thus the generated pH gradients. The solution to the continuous flow model, where the medium convection is considered, shows that liquid convection has significant effects on temperature distribution and the peak temperature location. PMID- 16703633 TI - Meta-analysis of prophylactic antibiotic use in acute necrotizing pancreatitis. AB - BACKGROUND: Death from infected necrosis in acute pancreatitis is common and prevention has focused on prophylactic antibiotics. This study assesses whether intravenous prophylactic antibiotic use reduces infected necrosis and death in acute necrotizing pancreatitis. METHODS: A meta-analysis of randomized controlled trials was carried out. Medline, Web of Science, the Cochrane controlled trials register and international conference proceedings were searched, with a citation review of relevant primary and review articles. RESULTS: Six of 328 studies assessed were included in data extraction. Primary outcome measures were infected necrosis and death. Secondary outcome measures were non-pancreatic infections, surgical intervention and length of hospital stay. Prophylactic antibiotic use was not associated with a statistically significant reduction in infected necrosis (relative risk (RR) 0.77 (95 per cent confidence interval (c.i.) 0.54 to 1.12); P = 0.173), mortality (RR 0.78 (95 per cent c.i. 0.44 to 1.39); P = 0.404), non-pancreatic infections (RR 0.71 (95 per cent c.i. 0.32 to 1.58); P = 0.402) and surgical intervention (RR 0.78 (95 per cent c.i. 0.55 to 1.11); P = 0.167). It was, however, associated with a statistically significant reduction in hospital stay (P = 0.040). CONCLUSION: Prophylactic antibiotics do not prevent infected necrosis or death in acute necrotizing pancreatitis. PMID- 16703635 TI - Association of peripheral total and differential leukocyte counts with metabolic syndrome and risk of ischemic cardiovascular diseases in patients with type 2 diabetes mellitus. AB - BACKGROUND: There is increasing evidence that leukocytes play a central role in obesity, glucose intolerance, type 2 diabetes mellitus (T2DM), and cardiovascular diseases, but the role of differential leukocytes in metabolic syndrome (MetS) and atherosclerosis is largely unknown. The aim of this study was to examine the relationship between the component features of MetS and peripheral leukocyte counts and to explore whether leukocyte counts are associated with clustering of MetS and macrovascular diseases in patients with T2DM. METHODS: 1872 subjects with T2DM who enrolled in a diabetes disease management program were studied. The definition of MetS was modified from that outlined by the criteria of NCEP-ATP III. Areas under the receiver-operating characteristic curve and odds ratios at various intervals of the WBC counts were computed. RESULTS: The peripheral total leukocyte, monocyte, and neutrophil cell counts are increased parallel to the clustering of components of MetS. When white cell counts were analyzed per quartile and as continuous variables after adjustment for age, sex, and other known risk factors with multiple regression analysis, peripheral total leukocyte, monocyte, neutrophils, and lymphocyte counts were independently and significantly associated with specific features of clustering of MetS and prevalence of ischemic cardiovascular diseases. Leukocyte counts, especially neutrophil/lymphocyte ratio, in addition with MetS is associated with the risk of ischemic cardiovascular diseases. CONCLUSION: Our results indicate that differential leukocyte counts are associated with MetS and that peripheral leukocytes may play a role in the pathogenesis of macrovascular complications in patients with T2DM. PMID- 16703637 TI - High-nuclearity, high-symmetry, high-spin molecules: A mixed-valence Mn10 cage possessing rare T symmetry and an S = 22 ground state. PMID- 16703636 TI - Mechanism of the vinylcyclobutane rearrangement of sceptrin to ageliferin and nagelamide E. PMID- 16703638 TI - A snapshot of P4 tetrahedron opening: Rh- and Ir-mediated activation of white phosphorus. PMID- 16703639 TI - Highly stereoselective synthesis of TMS-, alkyl-, or aryl-substituted cis [3]cumulenols via alpha-alkynylated zirconacyclopentenes. PMID- 16703640 TI - Radical photochemistry in oxygen-loaded ices. PMID- 16703641 TI - Multiple metal-carbon bonds for catalytic metathesis reactions (Nobel Lecture). PMID- 16703642 TI - [{(tBu2Me)2Si}3Li4]2-: An aggregated dianion of a 1,1-dilithiosilane with a unique structural motif. PMID- 16703643 TI - Formation of zirconocene fluoro complexes: No deactivation in the polymerization of olefins by the contact-ion-pair catalysts [Cp'2ZrR]+)[RB(C6F5)3]-. PMID- 16703644 TI - Insulin resistance in diabetic nephropathy--cause or consequence? AB - Insulin resistance (IR) is associated with multiple risk factors for cardiovascular disease. Many studies have shown that IR is present in chronic renal failure (CRF), and recent evidence suggests that IR can also occur in the early stages of renal disease. Patients with diabetic nephropathy (DN) have an increase in cardiovascular mortality, and since IR may be a contributing factor, this emphasizes the importance of a detailed understanding of the mechanisms linking IR and renal dysfunction at different stages of DN. IR can be detected early on in DN, e.g. at the stage of microalbuminuria (MA) and this could indicate a common genetic trait for IR and DN. As DN progresses further, IR is aggravated and it may, in addition to other factors, possibly accelerate the decline in renal function toward end-stage renal disease (ESRD). Several potentially modifiable mechanisms including circulating hormones, neuroendocrine pathways and chronic inflammation, are said to contribute to the worsening of IR. In ESRD, uremic toxins are of major importance. In this review article, we address the association between different stages of DN and IR and attempt to summarize major findings on potential mechanisms linking DN and IR. We conclude that IR is a consequence, and potentially also a cause of DN. In addition, there are probably genetic and environmental background factors that predispose to both IR and DN. PMID- 16703645 TI - Impact of oedema on recovery after major abdominal surgery and potential value of multifrequency bioimpedance measurements (Br J Surg 2006; 93: 354-361). PMID- 16703647 TI - Quantification of zolpidem in human plasma by high-performance liquid chromatography with fluorescence detection. AB - A simple, reliable HPLC method with fluorescence detection (excitation 320 and emission 388 nm) was developed and validated for quantitation of zolpidem in human plasma. Following a single-step liquid-liquid extraction, the analyte and internal standard (quinine) were separated using an isocratic mobile phase on a reversed-phase C(18) column. The lower limit of quantitation was 1.8 ng/mL, with a relative standard deviation of less than 5%. A linear dynamic range of 1.8-288 ng/mL was established. This HPLC method was validated with between-batch and within-batch precision of 1.7-4.8 and 1.2-2.3%, respectively. The between-batch and within-batch accuracy was 95.3-100.4 and 95.5-102.7%, respectively. Frequently coadministered drugs did not interfere with the described methodology. Stability of zolpidem in plasma was excellent, with no evidence of degradation during sample processing (autosampler) and 30 days storage in a freezer. This validated method is simple and repeatable enough to be used in pharmacokinetic studies. PMID- 16703648 TI - Inadvertent tracheal perforation during thyroidectomy (Br J Surg 2006; 93: 55 56). PMID- 16703650 TI - Bariatric surgery--a successful way to battle the weight crisis (Br J Surg 2006; 93: 259-260). PMID- 16703649 TI - Local recurrence after curative resection for rectal cancer is associated with anterior position of the tumour (Br J Surg 2006; 93: 105-112). PMID- 16703651 TI - The rapidly growing thyroid mass. PMID- 16703652 TI - Tissue engineering of vascular conduits. AB - BACKGROUND: Autologous conduits are not available in up to 40 per cent of patients with arteriopathy who require coronary or lower limb revascularization, and access sites for renal dialysis may eventually become exhausted. Synthetic prostheses achieve a poor patency rate in small-calibre anastomoses. This review examines how vascular tissue engineering may be used to address these issues. METHODS: A Medline search was performed, using the keywords "vascular tissue engineering", "small diameter vascular conduit", "vascular cell biology", "biomechanics", "cell seeding" and "graft endothelialization". Key references were hand-searched for relevant papers. RESULTS AND CONCLUSION: In vitro and in vivo approaches are currently being used for guided cell repopulation of both biological and synthetic scaffolds. The major clinical problem has been extended culture time (approximately 6 weeks), which precludes their use in the acute setting. However, recent advances have led not only to improved patency rates for prostheses, but also to a potential reduction in culture time. In addition, increased mobilization of endothelial progenitor cells in the presence of ischaemic tissue may increase the autologous cell yield for scaffold reseeding with further reduction in culture time. PMID- 16703653 TI - Randomized clinical trial of liver resection with and without hepatic pedicle clamping. AB - BACKGROUND: The purpose of this study was to compare the perioperative outcome of liver resection with and without intermittent hepatic pedicle clamping. METHODS: Between June 2002 and June 2004, 126 consecutive patients with resectable liver tumours were randomized to undergo resection with (63 patients) or without (63 patients) intermittent hepatic pedicle clamping. RESULTS: The transection time was significantly higher in the group without hepatic pedicle clamping. The blood loss per cm(2) was similar in the two groups: 2.7 ml/cm(2) in the group with versus 3.2 ml/cm(2) in group without hepatic pedicle clamping (P = 0.425). In the subset of patients with an abnormal liver, there were no differences in blood loss per transection surface: 3.1 ml/cm(2) in the group with versus 2.9 ml/cm(2) in the group without clamping (P = 0.829). The rate of blood transfusions was not higher in the non-clamping group. No differences were observed in the postoperative liver enzyme serum levels, the in-hospital mortality (one patient in each group) or the number of complications. CONCLUSION: This study showed clearly that liver resection without hepatic pedicle clamping is safe, even in patients with a diseased liver. PMID- 16703654 TI - An optically reversible switching membrane surface. PMID- 16703655 TI - Enantiomeric separation of fluoxetine derivatives on polysaccharide-based chiral columns. AB - A high performance liquid chromatography (HPLC) method employing amylose-based chiral columns (Chiralpak AD-RH and Chiralpak AD) and cellulose-based chiral columns (Chiralcel OD) as chiral stationary phases have been developed for the enantiomeric separation of fluoxetine (FLX) derivatives. The FLX was derivatized with 4-(N-chloroformylmethyl-N-methyl)amino-7-nitro-2,1,3-benzoxadiazole (NBD COCl) and 4-(N-chloroformylmethyl-N-methyl)amino-7-N,N-dimethylaminosulfonyl 2,1,3-benzoxadiazole (DBD-COCl), respectively. Influence of the mobile phase composition and column temperature on the enantioseparation was discussed during the separation. On the basis of separation of derivatized FLX enantiomers, the paper also discussed the separation mechanism on the chiral stationary phases used. PMID- 16703656 TI - Methods for assessing the toxicity of herbicides to submersed aquatic plants. AB - A new test design for the non-axenic submergent aquatic macrophytes Elodea canadensis Michx. and Myriophyllum spicatum L. has been developed for potential use in herbicide toxicity testing. For the non-axenic cultures, the best growth conditions were observed in the Elendt-M4 medium in which no growth of algae or bacteria was observed. Cuttings were placed in beakers containing only the artificial M4 medium or were planted in small beakers containing OECD (Organisation for Economic Cooperation and Development) sediment (5% peat, 75% sand, 20% kaolinite), which were then placed in larger vessels with the M4 medium. The plants were observed for main and secondary shoot length, biomass and root formation within 2-3 weeks of planting. Growth rates were calculated for total plant length and biomass. The variance between the replicates was low throughout the experiment [coefficient of variation (CV) < 26% for total plant length, and between 16 and 40% for biomass]. Relative growth rates based on total plant length were determined as 0.028 and 0.050 per day for M. spicatum in the systems containing M4 medium only and medium plus sediment respectively. Similar results were observed for E. canadensis, with relative growth rates of 0.26 and 0.073 per day in the two test systems. The root-shoot ratio at harvest was greater by a factor of 2-3 for E. canadensis in the M4 medium than in the system containing sediment. However, comparable ratios were observed for M. spicatum in the two test systems. Both growth in total plant length and growth in biomass of the two species have potential as measures of toxicity. PMID- 16703657 TI - Asymmetric amplification by kinetic resolution using a racemic reagent: example in amine acetylation. AB - The reaction of a racemic reagent on a mixture of enantiomers with small ee (ee=enantiomeric excess) has been studied for amine acylation. A substantial asymmetric amplification could be realized, for example, from 67 to >95.5 ee. The combination of asymmetric amplifications is subsequently discussed. Two sequential asymmetric amplifications, one using a racemic reagent and another using a positive nonlinear effect allowed us to start from 1.5 % ee and end with a large amount of a product of 97 % ee. PMID- 16703659 TI - New life for spinal cord stimulators. PMID- 16703658 TI - Synthesis, structural analysis, and visualization of a library of dendronized polyphenylacetylenes. AB - A library of eleven high cis-content cis-transoidal polyphenylacetylenes (PPAs) dendronized with self-assembling dendrons was prepared from a library of fifteen convergently synthesized macromonomers. Using [Rh(C triple bond CPh)(nbd)(PPh(3))(2)] (nbd=2,5-norbornadiene) in the presence of 10 equiv of N,N dimethylaminopyridine, predictive control over molecular weight and narrow molecular weight distribution are obtained. The PPA backbone serves as a helical scaffold for the self-assembling dendrons. The dendron primary structure dictates the diameter of the cylindrical PPAs in bulk, both in the self-organized hexagonal columnar (Phi(h)) lattice determined by X-ray diffraction (XRD) and in monolayers on highly ordered pyrolytic graphite (HOPG) and mica visualized by atomic force microscopy (AFM). Thermal and bulk phase characteristics of the cylindrical PPAs reinforces the generality that flexible polymer backbones adopt a helical conformation within the cylindrical macromolecules generated by polymers jacketed with self-assembling dendrons. PMID- 16703660 TI - Lower cholesterol without statins. Options are available. PMID- 16703661 TI - Does magnesium make bones stronger? My wife is 72 and she's taking magnesium supplements for her bones. PMID- 16703663 TI - Endocrine-related resources from the National Institutes of Health. PMID- 16703662 TI - Once a scientist, always a scientist. A double anniversary for A. Michael Michelson in 2006: 80th birthday and 60 years in research. PMID- 16703664 TI - Sensing inorganic carbon: CO2 and HCO3-. AB - Enzymes and transporters that catalyse reactions involving inorganic carbon are well characterized with respect to the species of inorganic carbon (CO2 or HCO3-) with which they interact. There is less information on the species recognized by proteins that sense inorganic carbon. In this issue of the Biochemical Journal, Hammer and colleagues show conclusively that cyanobacterial adenylyl cyclases are activated by CO2 and not HCO3-, as was believed previously. While in some circumstances a similar in vivo regulatory outcome is achieved from sensing HCO3- as from sensing CO2, there are cases in which the outcomes are significantly different. The most striking example is where a compartment lacks carbonic anhydrase yet supports large metabolic fluxes of inorganic carbon species so that CO2 and HCO3- are not at equilibrium. Other examples involve changes in pH, or temperature, of a compartment containing an equilibrium mixture of CO2 and HCO3-. PMID- 16703665 TI - Cleavage, a real turn-off? HIV-mediated proteolysis of PABP1. AB - In this issue of the Biochemical Journal, Alvarez and colleagues have identified PABP1 [poly(A)-binding protein 1] as a target of protease cleavage during HIV infection. The study shows that HIV-1, HIV-2 and mouse mammary tumour virus, but not other retroviruses, target PABP1 for cleavage and identifies cleavage sites within the RNA-recognition motifs and C-terminal region of the protein. This suggests that PABP1 cleavage may be important in the shut-off of host translation during HIV infection. This extends the viral families that are known to target PABP1 to include Retroviridae, suggesting that PABP1 may be a central target of viral infection. PMID- 16703666 TI - HIV Tat-mediated transcriptional regulation of proteasome protein cleavage specificity. AB - The major antigen-adapted immune response protecting a vertebrate against virus infection is that mediated by CTLs (cytotoxic T-lymphocytes). CTLs destroy virus infected cells, thereby containing the infection. They are activated by recognition of peptide antigens or epitopes, presented to them in the context of MHC I proteins. These epitopes are derived from proteolytic degradation of endogenously synthesized proteins, which is mediated by the proteasome. Augmentation of epitope presentation by MHC I is thought to be effected by the immunoproteasome, induced in response to IFN-gamma (interferon-gamma) in some cells, and constitutively expressed in others. In this issue of the Biochemical Journal, Remoli and colleagues describe the manipulation of the immunoproteasome by the Tat (transcriptional activation) protein of HIV. The authors show that Tat deregulates the balance of the three proteins, LMP2 (low-molecular-mass polypeptide 2), LMP7 and MECL1 (multicatalytic endopeptidase complex-like 1), which distinguish the immunoproteasome from the proteasome, and they provide a molecular explanation. Intracellular Tat sequesters IRF-1 (interferon-regulatory factor-1) from its cognate promoter element, where normally it associates with STAT1 (signal transducer and activator of transcription 1) to activate basal transcription of the LMP2 gene. LMP2 expression is inhibited as a consequence, skewing the stoichiometry of the immunoproteasome and changing its enzymatic activity. These findings provide a molecular account of an immunomodulatory activity of HIV: changing the peptide antigen profile of cells expressing or exposed to Tat. They may also provide an avenue for manipulating vaccine efficacy and specificity with Tat-based adjuvants. PMID- 16703667 TI - [Perioperative smoking. Conference of experts. Short text. 2005]. PMID- 16703668 TI - [41th Congress of EASD (European Association for The Study of Diabetes) 10 to 15 September 2005, Athens, Greece]. PMID- 16703669 TI - [First geriatric oncology colloquium Care of elderly cancer patients: an art, science and challenge Montreal, September 16th and 17th 2005]. PMID- 16703670 TI - Climate change and low temperature bioscience. PMID- 16703671 TI - Austria plans gym discounts to help citizens get fit. PMID- 16703672 TI - Application of a blood-brain-barrier-penetrating form of GDNF in a mouse model for Parkinson's disease. AB - Glial-cell-line-derived neurotrophic factor (GDNF) promotes mesencephalic dopaminergic neuronal survival in several in vitro and in vivo models. As the demise of dopaminergic neurons is the cause for Parkinson's disease (PD) symptoms, GDNF is a promising agent for its treatment. However, this neurotrophin is unable to cross the blood-brain barrier, which has complicated its clinical use. Therefore, ways to deliver GDNF into the central nervous system in an effective manner are needed. The HIV-1-Tat-derived cell-penetrating peptide (CPP) provides a means to deliver fusion proteins into the brain. We generated a fusion protein between the 11 amino acid CPP of Tat and the rat GDNF mature protein to deliver GDNF across the blood-brain barrier. We showed previously that Tat-GDNF enhances the neuroprotective effect of GDNF in in vivo models for nerve trauma and ischemia. Here, we tested its effect in a subchronic scheme of 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine (MPTP) application into the mouse as a model for PD to evaluate the effect of Tat-GDNF fusion protein in dopaminergic neuron survival. We showed that the fusion protein did indeed reach the dopaminergic neurons. However, the in vivo application of Tat-GDNF did not provide neuroprotection of dopaminergic neurons, as revealed by immunohistochemistry and counting of the number of tyrosine-hydroxylase-immunoreactive neurons in the substantia nigra pars compacta. Possibly, GDNF does protect nigro-striatal projections of those neurons that survive MPTP treatment but does not increase the number of surviving dopaminergic neurons. A concomitant treatment of Tat-GDNF with an anti-apoptotic Tat-fusion protein might be beneficial. PMID- 16703673 TI - Brain responses to a subject's own name uttered by a familiar voice. AB - Hearing one's own first name automatically elicits a robust electrophysiological response, even in conditions of reduced consciousness like sleep. In a search for objective clues to superior cognitive functions in comatose patients, we looked for an optimal auditory stimulation paradigm mobilizing a large population of neurons. Our hypothesis was that wider ERPs would be obtained in response to the subject's own name (SON) when a familiar person uttered it. In 15 healthy awake volunteers, we tested a passive oddball paradigm with three different novels presented with the same probability (P = 0.02): SON uttered by a familiar voice (FV) or by an unknown voice (NFV) and a non-vocal stimulus (NV) which preserved most of the physical characteristics of SON FV. ERP (32 electrodes) and scalp current density (SCD) maps were analyzed. SON appeared to generate more robust responses related to involuntary attention switching (MMN/N2b, novelty P3) than NV. When uttered by a familiar person, the SON elicited larger response amplitudes in the late phase of novelty P3 (after 300 ms). Most important differences were found in the late slow waves where two components could be temporally and spatially dissociated. A larger parietal component for FV than for NFV suggested deeper high-level processing, even if the subjects were not required to explicitly differentiate or recognize the voices. This passive protocol could therefore provide a valuable tool for clinicians to test residual superior cognitive functions in uncooperative patients. PMID- 16703674 TI - China's rural health reforms tackle entrenched inequalities. PMID- 16703676 TI - Romanian parents keep HIV a secret from infected children. PMID- 16703677 TI - Brief history of the Drug Evaluation Committee (DEC) and its role in the college on problems of drug dependence. PMID- 16703675 TI - The Fas gene A-670G polymorphism is not associated with sporadic Alzheimer disease in a Chinese Han population. AB - Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by excessive neuronal loss, intracellular neurofibrillary tangles and extracellular deposition of amyloid beta-peptide (Abeta). The Fas antigen is a cell surface receptor-mediating cell apoptosis. Several lines of evidence have made Fas/Fas ligand induced apoptosis play an important role in the pathogenesis of AD. Moreover, the Fas gene is located on chromosome 10q24.1, a region of linkage to late-onset AD. Several reports have investigated the association between a single nucleotide polymorphism (SNP) that is located at position -670 of Fas gene and AD, but yielded ambiguous results. To figure out the association of this SNP with sporadic AD in Chinese Han population, we have analyzed 509 patients with AD and 561 controls for the genetic association studies. Our results indicate that the distribution of the Fas genotypes (chi(2) = 0.66, P = 0.72) and alleles (chi(2) = 0.70, P = 0.40) did not differ significantly. The similar results were observed when AD and control groups were stratified by age/age at onset and sex (P > 0.10). The present data revealed no significant effect of the genotypes on the age of onset for developing AD, and no significant association between the genotypes and the severity of the disease. PMID- 16703678 TI - [Rupture of the superficial dorsal vein of the penis]. AB - Rupture of the superficial dorsal vein of the penis during intercourse is an unfrequented entity that makes the differential diagnosis with other acute penile injuries that may require surgical exploration necessary. We report the case of a 58-year-old male patient with 24-hours evolution painless haematoma after intercourse; its evolution and characteristic physical exploration enable us to adopt a conservative approach that resulted in complete recovery without sequels. PMID- 16703680 TI - [Evaluation of drug-induced lung disease--guideline for the treatment]. PMID- 16703679 TI - Sexual conflict: a new paradigm? Papers from a discussion meeting at the Royal Society. May 9-10, 2005. London, United Kingdom. PMID- 16703681 TI - [The 79th annual meeting of the Japanese Society for Bacteriology. Kanazawa, Japan. March 29-31, 2006. Abstracts]. PMID- 16703682 TI - Endovascular exclusion of symptomatic bilateral common lliac artery anueurysms with preservation of an aneurysmal internal lliac artery via a reverse-U stent graft. AB - PURPOSE: To report a technique to maintain flow to an aneurysmal internal iliac artery (IIA) when treating bilateral common iliac artery (CIA) aneurysms with an aortomonoiliac stent-graft and femorofemoral bypass. TECHNIQUE: First, an external iliac artery (EIA) to IIA endograft is placed distal to the IIA aneurysm then the contralateral IIA is embolized with coils. An aortomonoiliac stent-graft extending to the contralateral EIA is placed, and the procedure is completed with a femorofemoral bypass. CONCLUSION: Endovascular treatment of bilateral CIA aneurysms and combined with a unilateral IIA aneurysm is a technically demanding procedure. An endovascular repair with retrograde (reverse-U stent-graft) hypogastric artery preservation can be considered a first choice until the use of branched iliac stent-grafts becomes more widespread. PMID- 16703683 TI - The nature of plant growth-promoting effects of a pseudoalteromonad associated with the marine algae Laminaria japonica and linked to catalase excretion. AB - AIMS: The goal of this study was to identify a marine algae-associated bacterium isolated from Laminaria japonica and investigate this microorganism's growth promoting effects on plants. METHODS AND RESULTS: The bacterium, identified as Pseudoalteromonas porphyrae, was determined to display a biostimulatory activity for seed germination and shoot growth in several agricultural plants and also for growth in ginseng callus cell culture. This biostimulatory activity was linked to a catalase enzyme that was excreted in the maximal amount during the transition from logarithmic growth phase to stationary growth phase. In addition, selected shifts in growth temperature and medium salinity affected the amount of enzyme excreted. The purified catalase was determined to be composed of identical subunits. The catalase of interest displayed significantly higher biostimulatory activity than the catalase from bovine liver. CONCLUSIONS: The catalase investigated in this study is unique in that it promotes growth in and possibly contributes to stress tolerance of plants. SIGNIFICANCE AND IMPACT OF THE STUDY: The catalase of interest has the potential for use in treatments that aim to improve percent seed germination as well as obtaining tall shoots in a shorter time period. PMID- 16703684 TI - Comment on "Patient attitudes to sternotomy and thoracotomy scars" by Crossland et al. Thorac Cardiovasc Surg 2005; 53: 93-95. PMID- 16703685 TI - Telephone interview with Fresenius Medical Care project leader Mr. Jonas: launch of 5008 Therapy System. PMID- 16703686 TI - Summing it up: Collective lessons from the experience of seven coalitions. PMID- 16703687 TI - Extracorporeal high intensity focused ultrasound in the treatment of patients with solid malignancy. AB - The ideal treatment of localized cancer should cause the complete death of tumor cells without damage to surrounding normal tissue. High intensity focused ultrasound (HIFU) is such a potential treatment, which can induce complete coagulation necrosis of a targeted tumor, at depth, through the intact skin. The concept of using HIFU as a non-invasive therapy has attracted attention in medicine for 60 years. Recently, it has received increasing interest as a promising modality for the treatment of localized solid malignancies. The goal of this article is to introduce recent clinical developments in the use of extracorporeal HIFU ablation for solid tumors, including those of liver, breast, bone, kidney, pancreas, soft tissue, and uterus. It describes the physical principles and ablative mechanisms, three-dimensional therapeutic regimes, and medical imaging used in HIFU. Currently, large numbers of patients with solid malignancy are already treated using HIFU, and short-term clinical results are very encouraging. However, large-scale randomized clinical trials are necessary to evaluate long-term efficacy of HIFU treatment for solid malignancies. It is concluded that this non-invasive ablation can be considered as a conventional therapy for widespread clinical use only when the results from prospective, randomized clinical trials worldwide are available. PMID- 16703688 TI - [12th Parizek Seminar. An international conference on prevention of venous thrombosis and pulmonary embolism sponsored by the Czech Hematologic Society CLS JEP on 16-17 March 2006 in Novy Jicin, Czech Republic]. PMID- 16703689 TI - Don't ignore the right radial approach. PMID- 16703690 TI - Judging fiduciary duty. AB - How much oversight of fiduciary duty is enough? The stakes are higher than ever for trustees to make certain they know what the term truly means. PMID- 16703691 TI - Knowledge building: how to raise your board's IQ. AB - As the complexities of health care increase, so does the board's need to stay current on a growing list of sophisticated topics. PMID- 16703692 TI - Trustee workbook 2. The challenges of system governance. PMID- 16703693 TI - Strategies for the hospital workforce of 2010. AB - Dramatic shifts in medical science, technology, demographics and economics will require that the health care labor force evolve along with changes in the delivery system. PMID- 16703694 TI - We're listening: a Center progress report. PMID- 16703695 TI - Stuart Lang: building one city out of two. PMID- 16703696 TI - The Patriot Act: violating trustees' trust? PMID- 16703697 TI - Director fitness: a sensitive issue requires a serious response. PMID- 16703698 TI - The military management of burn injury--some historic reflections. PMID- 16703699 TI - Exercise Maganga Mercury 23 January-10 February 2006. AB - It is very likely that Royal Navy doctors will be called upon more and more in the future to work on similar overseas exercises. I found the experience immensely challenging and educational and would have no hesitation in volunteering for further deployments or recommending other RN doctors to do the same. PMID- 16703700 TI - In response to Dr R Agnew's letter to the editor JRNMS vol 91 No 3 2005. PMID- 16703701 TI - Subject: Re: John Pollard, 1787-1868. PMID- 16703702 TI - Professional liability insurance carriers: friend or foe? PMID- 16703703 TI - Loser pays laws are necessary to deter frivolous lawsuits. PMID- 16703704 TI - Missouri Medicaid reform...what can physicians do? PMID- 16703705 TI - "People need a fairy tale": the embryonic stem cell and cloning debate in Missouri. PMID- 16703706 TI - On medical futility: considerations and guidelines. PMID- 16703707 TI - The future of electronic medical records--part I. PMID- 16703708 TI - Malpractice from A-Z. Part 2: Being sued? What to do and what to expect. PMID- 16703710 TI - Teachable moments: how combining the medical library and CME can increase quality and decrease overhead. PMID- 16703709 TI - CA-MRSA: An old foe develops new fangs. PMID- 16703711 TI - Funding for medicine at UMC would benefit all Missourians. PMID- 16703712 TI - Defining success at the University of Missouri Department of Family and Community Medicine. PMID- 16703713 TI - Family medicine update 2006 at the University of Missouri-Columbia School of Medicine. PMID- 16703714 TI - Improving care at lower cost for end-stage heart and lung disease: integrating end of life planning with home care. AB - Patients with advanced heart and lung disease experience exacerbations resulting in hospitalizations and interventions the patient may not desire. Strategies are needed that address end of life issues, honor preferences, and improve care without increasing cost. This study examines the impact on hospitalization and care cost of an integrated system of end of life care and interdisciplinary home care for mid-Missouri veterans with advanced congestive heart failure or chronic obstructive pulmonary disease. PMID- 16703716 TI - Comprehensive geriatric assessment for non-geriatricians. AB - Comprehensive geriatric assessment (CGA) consists of medical history taking, medication review, physical examination with special attention to sensory and neurological evaluation, and use of assessment instruments to measure cognitive, affective, functional, social, and economic status. CGA is most often beneficial when applied to frail elders at risk of functional decline. Beneficial outcomes may include decreased risks of falls, cognitive decline, and nursing home placement--in addition to improved sense of well-being and satisfaction with care. PMID- 16703715 TI - Marijuana and other illicit drug use and the risk of injury: A case-control study. AB - OBJECTIVES: Assess the associations between marijuana or other illicit drug use and injury risk. METHODS: Case-control study. Cases were emergency department patients with acute injury. Age-sex matched controls were interviewed by telephone. RESULTS: Marijuana use in the past seven days was associated with decreased risk, and use of other illicit drugs was associated with increased risk. Controlling for confounding had little effect. CONCLUSIONS: Marijuana use may be associated with a decreased risk of injury. Other illicit drug use was associated with increased risk. PMID- 16703717 TI - Brief hospital quality improvement program: preventing venous thromboembolism. AB - Venous thromboembolism (VTE) is a frequent but potentially preventable complication in the hospitalized patient. Solid medical evidence exists about pharmacological and other techniques of effective prevention. As part of a University Hospital quality improvement (QI) program, the Department of Familyand Community Medicine (FCM) increased the rate of prevention from 25 percent to more than 90 percent of eligible patients in just eight months. This overview describes the steps taken in this project to achieve rapid results using accepted QI methodology to reduce VTE. PMID- 16703718 TI - Recurrent vulvovaginal candidiasis. AB - Recurrent vulvovaginal candidiasis affects five percent of women of child-bearing age. The most common organism is Candidia albicans, but an increasing number of infections are caused by nonalbicans species. Fungal culture directs treatment as nonalbicans species may be azole resistant. C. albicans will respond to anyazole antifungal. Treat C. glabrata with boric acid. Maintenance therapy should be started immediately after treatment of the acute episode and should last for six months. PMID- 16703719 TI - Approach to shoulder pain in primary care. AB - Shoulder pain is a common presenting complaint in primary care. A full understanding of the spectrum of possible shoulder disorders can be daunting for many primary care physicians, but a practical approach towards conditions which are most common in primary care can allow for the initial management of most of these conditions. Referral for a concentrated physical therapy program can improve several of the common conditions, even if the precise diagnosis remains elusive. PMID- 16703720 TI - Addressing the insidious health and financial burden of tobacco use in Missouri: the role of medical and public health professionals. AB - Despite the widespread knowledge that tobacco use causes death and debilitating disease, tobacco control efforts in Missouri remain inadequate. No other health issue combines the prevalence, lethality, and neglect of tobacco addiction. This paper discusses comprehensive tobacco-use prevention and cessation programs that could, if implemented, effectively reduce tobacco-related health problems. Successful tobacco control efforts change society by creating an environment in which nonsmoking is the norm and quitting smoking is supported. This paper concludes that it is time for medicine and public health professionals to communicate, collaborate, and assume leadership in reducing the health burden of tobacco use in Missouri. PMID- 16703721 TI - Characteristics of smoking cessation guideline use by primary care physicians. AB - This study, based on a random sample mail survey of Missouri primary care physicians, identifies: (1) the extent and predictors of these physicians' awareness, agreement, adoption, and adherence to tobacco cessation guidelines; and (2) their knowledge/ beliefs about tobacco interventions. While primary care physicians play a key role in helping patients quit smoking, guidelines are not widely followed. "Reliance on respected colleagues for advice" was the one variable consistently identified as a predictor of guideline compliance. PMID- 16703722 TI - Modern surgical and interventional options for the management of heart failure. AB - Current standard drug therapy provides only palliative and symptomatic relief for patients with severe heart failure, but there is now a wide array of device applications that holds great promise for reducing the mortality and disability that result from heart failure. Patients with severe chronic and refractory heart failure should be considered for heart transplantation or mechanical circulatory supportwith a ventricular assist device. Newer surgical and interventional treatments are in continuous evolution. PMID- 16703723 TI - [Da Vinci advanced robotic laparoscopic surgery: origin and current clinical application in urology, and comparison with open and laparoscopic surgery]. AB - Over the last decade, open surgery, which is able to perform large extirpations and repairs of fragile tissues, is gradually being substituted with laparoscopic surgery due to the high benefits the latter entails for the patients, an also due to the learning difficulties for surgeons who must make up for such deficiencies applying higher efforts and a larger amount of stress. Robotic surgery stands in for the limitations of conventional laparoscopic surgery by means of performing more ergonomic and more accurate surgeries, particularly in the case of the most complex and difficult to access operations, such as radical prostatectomy. This review will perform a reminder of the history and clinical applications of new advanced and robotic technologies, and also a comparison with open surgery and conventional laparoscopy. PMID- 16703724 TI - [Determination of free prostatic specific antigen cut point for the selection of patients in first prostate biopsy]. AB - OBJECTIVES: Determine the cut point of free PSA rate for optimize the first prostate biopsy indication. MATERIAL AND METHODS: Prospective trial between june 2002-september 2004, We included patients in first prostate biopsy with normal rectal digital examen, total PSA between 3-10 ng/ml and normal transrectal prostate ultrasound. We realize descriptive stadistic analisis of variables age, total PSA, prostate volume and % free PSA and analitic stadistic analisis with ROC curves of variables total PSA and % free PSA for determine as of her predicts the best one rate of prostate cancer. RESULTS: We reclute 727 men with a mean age 62.91 years, total PSA mean 6.12 ng/ml, prostate volume mean 42.78 cc and % free PSA mean 15.22%. We had 106 prostate cancer, the prostate cancer rate in first biopsy was 14.6%; 77 cases had 1 lobe prostate cancer and 29 in 2 lobes. The most frecuent gleason was 6 (46 cases 43.4%) and the second gleason 7 (43 cases 40.6%). In the ROC curves analysis, total PSA had area under the curve 0.476 (p=0.3) and 0.611 (p=0.023) for % free PSA. The optime cut point for % free PSA in our trial was 19% (Sensibility 91.4% and Specificity 20%). The use of this cut point had allowed the saving us 138 biopsies (19.11%) with the lost one of diagnose of 10 cases of cancer of prostate. CONCLUSIONS: The use of the % free PSA is useful and allows in our region the best indication of the patients who are going to first biopsy of prostate, avoiding the accomplishment of unnecessary biopsies. PMID- 16703725 TI - [The utility of artificial neural networks in the prediction of prostate cancer on transrectal biopsy]. AB - OBJECTIVE: To determine whether the development of an artificial neural network (ANN) made up of clinical variables allows for the prediction of prostate biopsy (PB) outcome. MATERIALS AND METHODS: Patients (n=953) underwent PB at the Arquitecto Marcide Hospital in Ferrol (Spain), between january 2000 and june 2005. The variables studied were age, PSA, digital rectal examination (DRE) and prostate volume, data for all of which were available in 843 cases. In order to determine factors related to prostate cancer (PC) diagnosis, a logistic regression analysis and a feed-forward neural network were developed, including three hidden layer nodes and an output node, representing the probability of PC. Both models were constructed from a random sample of n=643 patients (derivation set). The predictive capacity was assessed with the remaining 200 patients (validation set), by means of ROC curves and the area under the curve (AUC). RESULTS: PC was detected in 500 (59.3%) cases. Adjusting for age, PSA, digital rectal examination and prostate volume, in a multivariate logistic regression model it was observed that all the variables were independent predictors of PC. The AUC were 0.693 for PSA, 0.707 for prostate volume, 0.815 for logistic regression and 0.819 for ANN. The predictive capacity of the ANN was significantly higher than that of the PSA (p=0.002) and prostate volume (p<0,001) and similar to that of logistic regression (p=0.760). CONCLUSIONS: The ANN shows a PC prediction capacity that is significantly higher than unimodal diagnosis methods, and similar to that of logistic regression. PMID- 16703726 TI - [Development of tissue microarray technology (TMA) for immunohistochemical study of molecular expression profiling in prostate cancer (part 1)]. AB - Tissue microarray technology (TMA) is nowadays considered as a powerful tool for the high-throughput analysis of molecular expression pattern of cancer. In this manuscript we show the experience of both groups in the design and building of a TMA for the study of protein expression pattern of prostatecancer as well as a summary of the technical points to analyze the results obtained with this technology. Today, different data generated by the immunostained tissues are studied to achieve a molecular profile in different clinical scenarios. PMID- 16703727 TI - [Clinicopathological study of incidental cancer prostate in patients undergoing surgery for symptomatic diagnosis of BPH]. AB - OBJECTIVES-INTRODUCTION: Retrospective study of a series of patients with prostate surgery (suprapubic prostate adenomectomy -APS-, and prostate transurethral resection -RTU-P-) for presumed BPH symptomatic non-respondent to the conservative medical treatment. Analysis of prevalence, incidence, clinical pathological, treatment, tumor progression and evolution of the patients with incidental prostate cancer (CPI) detected. MATERIAL AND METHODS: 1593 patients with prostate surgery (APS and RTU-P) during 6 years (1996-2001) were revised. APS 35%, RTU-P 65%. Revision of all pathological anatomy of surgical specimens and the evolutions of the patients with CPI. RESULTS: 78 CPI; Prevalence 4,89%; Incidence 13 cases/year. Mean age 73.6 years. Digital rectal examination was normal in 100%, mean PSA 6 ng/ml (0.5-30). Group APS: 25 CPI (32%); prevalence 4.55%; incidence 4 cases/year; mean PSA 7.7 ng/ml (2.8-30); mean weight resection 65 gs. Group RTU-P: 53 CPI (68%); prevalence 5,07%; incidence 9 cases/year; mean PSA 5.2 ng/ml (0,5-29); mean weight resection 20 gs. 22% biopsy previously by high PSA, mean PSA 14 ng/ml (4,8-30). Gleason average 5 (mean 4.8), rank 3-8. pTla 66%, pTlb 33%. TREATMENT: 57% follow-up watched without treatment (wait and see); 18% hormonal treatment; 3% finasteride; 9% Radical Prostatectomy; 9% radiotherapy. Follow Lost 4%. Mean follow-up 47.19 months (12-96). Tumor progression 13.3% (10 patients). Specific CPI mortality 2.6% (2 patients). CONCLUSIONS: We didn't observe significant differences between the prevalence of CPI in both groups (APS and RTU-P). The detected tumours were mainly well differentiated and in stage pTla. In more than half of the cases an expectant attitude without treatment was decided. 13,3% of tumor progression after 47.19 months of follow mean and specific CPI mortality 2.6%. PMID- 16703728 TI - [Tumor marker tests in bladder cancer]. AB - The gold standard for detecting bladder cancer is cystoscopy which identifies nearly all papillary and sessile lesions. However, it is an invasive procedure causing some discomfort for patients. Urine cytology is the standard non-invasive marker with very high specificity, but unfavourable poor sensitivity for Ta, G1, and T1 bladder tumors. To improve early detection of bladder cancer as well as to monitor treatment response and tumor recurrence, bladder tumor markers are eligible. An ideal bladder cancer test would have the potential to replace or delay cystoscopy in the follow-up of bladder cancer patients. In recent years, the FDA approved non-invasive tumor marker tests ImmunoCyt / uCyt+, BTA TRAK, BTA stat, NMP22, NMP22 BladderChek, and UroVysion have been investigated. The tests demonstrated higher sensitivity for diagnosis of bladder cancer compared to urine cytology. Overall, the mean sensitivity and mean specificity was 64-80% and 71 95% and the mean positive and negative predictive values to detect malignancy were 49-84% and 79-95%, respectively. BTA TRAK, BTA stat, NMP22, and NMP22 BladderChek assays are limited by false-positive results in patients with benign urological diseases such as hematuria, urocystitis, renal calculi or urinary tract infections. Due to low specificity BTA TRAK, BTA stat, NMP22, and NMP22 BladderChek should not be used without first ruling out benign or malignant genitourinary disease other than bladder cancer. With the exception of UroVysion achieving 80% sensitivity and 94% specificity, none of these non-invasive tests revealed a high sensitivity and specificity at the same time, which is a main demand to be made on an ideal tumor marker. Insufficient sensitivity along with limited specificity does not allow replacing cystoscopy in diagnosis of bladder cancer or treatment decisions based on a positive test result. PMID- 16703729 TI - [Endoscopic treatment of the benign prostatic hyperplasia with local anesthesia and sedation in patients with high surgical or anaesthesical risk]. AB - INTRODUCTION AND OBJECTIVE: [corrected] The aging of the population causes an increase of the number of men affected by benign prostatic hyperplasia (BPH) with a large number of pathology associated that may limit its surgical treatment. In this study we try to show the effectiveness and security of local anaesthesia and sedation in the endoscopic treatment of the BPH in patients with high surgical anaesthetic risk. METHODS: During the last 4 years, 38 patients underwent surgical treatment of BPH with ages between 78 and 86 years (average 81,4 +/- 3,2) with severe obstructive prostatic syndrome and high surgical-anaesthetic risk. 15 patients underwent, transurethral incision of prostate (TUIP) and in 23 patients, transurethral resection (TURP) was performed. The blockade anesthetic was made by transrectal periprostatic infiltration finger-guided with 10-15 millilitres of lidocaine 2%. During the procedure an intravenous perfusion of propofol and remifentanil was administered. The obtained results as well as the patient's satisfaction degree was evaluated by means of the elaboration of a questionnaire. RESULTS: The average length of the surgical treatment was of 12 +/ 5 minutes (rank 8 - 25), having itself not stated complications derived from the anesthetic technique. They referred a slight annoyance 4 patients (11%) that was treated increasing the rate of the sedative perfusion. The satisfaction's degree with the received treatment has been good in the 95% of patients. CONCLUSIONS: We considered that the TUIP as much as TURP made under local anesthesia with sedation are safe, effective methods and well tolerated, that fundamentally are indicated in aged patients and when the surgical-anesthetic risk is elevated. PMID- 16703730 TI - [Assessment of effectiveness and security of bladder neck cerclage after transvesical prostatectomy]. AB - INTRODUCTION: The Bladder neck cerclage diminishes the risk of bleeding after transvesical prostatectomy, but it increases the risk of suffering postoperative bladder neck sclerosis. Our objective is to value the effectiveness and therapeutic security of the temporary bladder neck cerclage after transvesical prostatectomy. MATERIAL AND METHODS: It was carried out a prospective and longitudinal study in a cohort of 25 age patients mean age 68,5 years (standard deviation, 2,6 years), subjected to transvesical prostatectomy (Freyer) with bladder neck cerclage using polipropilene number 1 suture, that was retired at the 24 hours of the surgery. To value the hemostatic utility of bladder neck cerclage, it was measured the hematocrit and hemoglobin concentration at the 24 hours of the intervention. To evaluate the appearance of obstructive sequels, the maximum urinary flow was measured three months after the surgery. RESULTS: It was observed a mean hematocrit at the 24 hours of surgery of 7,3%, and a mean decrease of the hemoglobin at the 24 hours of the surgery of 2,7 gr/dl. In any cases the postoperative hemoglobin concentration was inferior to 8 mg/dl, therefore, it was not necessary transfusion. The uroflowmetry carried out at 3 months of surgery showed that 24 of the 25 intervened patients presented a maximum flow superior to 15 ml/sg. CONCLUSIONS: The temporary bladder neck cerclage is a good hemostatic technique. The precocious cercalge retreat avoids the late sequels (bladder neck sclerosos), that originates the permanent cerclage. PMID- 16703731 TI - [Unusual elements in renal calculi]. AB - A group of 54 renal calculi were spontaneously passed renal stone after a nephritic colic. Two groups of calculi were found: papillary and non-papillary calculi. All calculi were analyzed by infrared spectroscopy and electronic microscopy scan (EMS) and EDAX. When the stones were analyzed with EDAX, elements such as C, N, O, Na, S, Mg, Al, Si, Cl, K, Ca, Mn, Fe, Ni, Zn were detected. The possible origin of these elements is discussed in this work. PMID- 16703732 TI - [Minimal invasive surgery in female urinary incontinence: TVT-O]. AB - OBJECTIVE: We presented our experience in the application of Tvt-O, a surgical a new technic for SUI. METHOD: Between April and December 2004, 96 patients underwent Tvt-O surgery, in Uroginecology and Vaginal Surgery Unit of the Gynecology and Obstetrics Department, Las Condes Clinic. Median age was 54 years old, weight 65 kgs. Urodinamia test: SUI II 80 cases, SUI III 4, SUI 0 in 3, and mixed urinary incontinence 9. Gynecare TVT Obturator System technique was used, Ethicon Johnson & Johnson. It has three specific instruments: helical passers, plastic tubes fixed to mesh of prolene, and a guide for the introduction of the needle. RESULTS: Media time was 7 minutes (4 to 15). Other gynecological surgery was associated in 77 (80%). Complications did not appear during the surgical act. No injury to bladder and urethra was observed. We observed in immediate postoperative period only one complication (1.04%). It was urinary retention. One complication appeared (1.04%) in remote postoperative period, corresponding polypropylene tape exposition in the anterior vaginal wall. SUI solution was obtained in 100% of patients. The medium time of observation is 6 months, 51 cases have more than 6 months of observation. DISCUSSION: Tvt-O is a surgical SUI correction technique with promissory results. Like the other types of TOT, their effectiveness will have in the long term to be evaluated. PMID- 16703733 TI - [Tadalafil vs sildenafil patient preference in Spanish men with erectile dysfunction: results from an International Multicentric Study]. AB - OBJECTIVE: To compare patient preference for sildenafil citrate (sildenafil) vs. tadalafil and for their respective dosing instructions in a cohort of Spanish patients with erectile dysfunction (ED). MATERIAL AND METHODS: Sixty four Spanish patients from a multicenter, two period, cross-over, double-blind study (265 patients enrolled in total) were randomized to receive on-demand sildenafil 50 mg or tadalafil 20 mg for 12 weeks and afterwards were crossed over to the alternate regimen for another 12 weeks to assess drug preference in an extension period of the study. Similarly, to evaluate preference for their respective dosing instructions, 30 patients were randomized to one of the 2 arms treated with tadalafil: one with sildenafil (S) dosing instructions and the other with tadalafil (T) dosing instructions. RESULTS: Seventy percent of 56 patients completing the study chose to receive tadalafil treatment versus sildenafil treatment (30%) in the extension period (p<0.01). Correspondingly, 73% of 13 evaluating each drug dosing instructions preferred T dosing instructions (p>0.05). Preference did not vary with age, concomitant diseases and previous use of sildenafil. CONCLUSIONS: In this study, 7 out of 10 patients preferred tadalafil and its dosing instructions to sildenafil, for the treatment of their ED. PMID- 16703735 TI - [Female paraurethral cysts. Experience of 4 cases]. AB - Parauthreal cysts are an uncommon pathology. We present 4 cases diagnosed and treated in our hospital during the last 10 years. PMID- 16703734 TI - [Trombosis of the dorsal penis vein (of Mondor's phlebitis). Presentation of a new case]. AB - OBJECTIVE: We present a new case of trombosis of the superficial dorsal penis vein called Penile Mondor's disease. The characteristics of the disease are reviewed and the most usual diagnostic and therapeutic methods. METHODS: The case of a 41 year old man is reviewed who consulted for pain and induration on the proximal part of the penis. RESULTS: After phisical examination and Eco-doppler was made the diagnosis of Mondor's disease. He receibed treatment with non steroidal antiinflamatories and antibiotics. CONCLUSIONS: The dorsal vein thrombosis is a rare disease with pain an induration of the dorsal part of the penis. The ethiology can be traumatic, neoplasic, excesive sexual activity or abstinence. Is necesary the diferencial diagnosis with esclerosant linphangitis and the most important imaging is the Eco-doppler. The treatment is based in non steroidal antiinflamatories and antibiotics wit infection. The local aplication of heparine can be useful and the surgery with thrombectomy and resection is for persistent cases. PMID- 16703736 TI - [Laparoscopic treatment of ovarian vein syndrome]. AB - OBJECTIVE: We report a case of Ovarian Vein Syndrome, describe its clinical symptoms and discuss its diagnosis and management including laparoscopic surgery treatment. MATERIALS AND METHODS: A 36-year-old female with right kidney recurring pain was studied by means of abdominal RX, urography, CT, MRI and ultrasonography and finally diagnosed from Ovarian Vein Syndrome. The case was resolved with laparoscopic surgery. CONCLUSIONS: Ovarian Vein Syndrome is an uncommon disorder. Differential diagnosis must be done with external processes that cause ureteral obstruction. Surgery is the first line treatment when clinical symptoms are present and, in our opinion,laparoscopic surgery is the best approach to treat this pathology. PMID- 16703737 TI - [Verrucous carcinoma of the penis arising from a lichen planus. A true preneoplastic lesion?]. AB - INTRODUCTION: Up to 30% of squamous cell carcinomas of the penis arise from a lichen sclerosus, but very few reports in which lichen planus was the preexisting lesion have been published. We report a male with verrucous carcinoma of penis that developed in an area of lichen planus. CLINICAL CASE: A 53-year-old male presented with an exophytic tumor on the glans penis that had been present for about 6 months. The lesion developed in a previously biopsied area of lichen planus hypertrophicus. After excision, histological diagnosis was verrucous carcinoma. COMMENT: The development of any subtype of squamous cell carcinoma of penis in a lichen planus can be coincidental, or a neoplastic transformation of lichen planus can take place. PMID- 16703738 TI - [The paraurethral cyst or Skene's duct cyst in the female newborn]. AB - The paraurethral cyst or Skene's duct cyst is a rare congenital anomaly in the female newborn. It appears like a round, yellow or orange-colored cystic mass on either side of the urethral meatus. The reported incidence is 1 in 2.000 to 7.000 female births. The low frequency of the Skene's duct cyst and the little awareness of it and its benignity may lead to wrong diagnosis that underestimates its incidence. There are authors that agree with surgical treatment of paraurethral cyst to obtain an early resolution, even though a majority agrees to manage conservatively. PMID- 16703739 TI - [Metastasic tumors in penis]. PMID- 16703740 TI - [Letter to the editor in refer to the article: High risk disseminated prostate cancer (consensus paper and recommendations)]. PMID- 16703741 TI - [Multiple lithiasis in a horseshoe kidney]. PMID- 16703742 TI - [Acute rejection in renal implant for echography]. PMID- 16703743 TI - [Gangrene of Fournier with urethral involvement: urethral reepitelization with conservative treatment]. PMID- 16703744 TI - The inaugural Frank Ellis Lecture--latrogenic cancer: the impact of intensity modulated radiotherapy. AB - It is an honour and personal pleasure to give the inaugural Frank Ellis Lecture to celebrate his 100th birthday, and to acknowledge his enormous contributions to radiation oncology. Intensity-modulated radiotherapy (IMRT) allows dose to be concentrated in the tumour volume while sparing normal tissues. However, the downside to IMRT is the potential to increase the number of radiation-induced second cancers because more fields are used which involves a bigger volume of normal tissue exposed to lower doses. It has been estimated that IMRT may double the incidence of solid cancers in long-term survivors. This may be acceptable in older patients if balanced by an improvement in local tumour control and reduced toxicity. On the other hand, the incidence of second cancers is higher in children, so that doubling it may not be acceptable. IMRT represents a special case for children. First, they are more sensitive to radiation-induced cancer than adults. Second, radiation scattered from the treatment volume is more important in the small body of the child. Third, there is the question of genetic susceptibility, as many childhood cancers involve a germline mutation. The levels of leakage radiation in current Linacs can be reduced, but the cost would be substantial. An alternative strategy is to replace X-rays with protons. This is an advantage only if the proton machine uses a pencil scanning beam, as passive modulation of a scattering foil produces neutrons, which results in an effective dose to the patient higher than that characteristic of IMRT. PMID- 16703745 TI - Explaining socioeconomic status effects in laryngeal cancer. AB - INTRODUCTION: People with lower socioeconomic status (SES) experience shorter survival times after a cancer diagnosis for many disease sites. We determined whether area-level SES was associated with the outcomes: cause-specific survival and local-regional failure in laryngeal cancer in Ontario, Canada. When we found an association we sought explanations that might be related to access to care including age, sex, rural residence, tumor stage, lymph node status, use of diagnostic imaging, treatment type, percentage of prescribed radiotherapy delivered, number of radiotherapy interruption days, treatment waiting time, and treating cancer center. MATERIALS AND METHODS: The study population consisted of 661 glottic and 495 supraglottic stage-stratified randomly-sampled patients identified using the Ontario Cancer Registry. Area-level SES quintiles were assigned using adjusted median household income from the Canadian Census. Other data were collected from patient charts. Explanations for SES effects were determined by measuring whether the effect moved toward the null value by at least 10% when an access indicator was added to a the model. RESULTS: Socioeconomic status was not related to either outcome for those with supraglottic cancer, but an association was present in glottic cancer. With the highest socioeconomic status quintile as the referent group, the relative risks for patients in the lowest socioeconomic quintile were 2.75 (95% CI 1.48, 5.12) for cause-specific survival and 1.90 (95% CI 1.24, 2.93) for local-regional failure. Disease stage as measured by T-category explained between 3% and 23% of these socioeconomic effects. None of the other access indicators met our 10% change criterion. CONCLUSION: We question why people in lower socioeconomic quintiles were not diagnosed earlier in the disease progression. Having ruled out several variables that may be related to access to care, additional biologic and social variables should be examined to further understand socioeconomic status effects. PMID- 16703746 TI - Dose escalation of three-dimensional conformal radiotherapy for locally recurrent nasopharyngeal carcinoma: a prospective randomised study. AB - AIMS: To investigate prospectively the feasibility and efficacy of dose escalation using three-dimensional conformal radiotherapy (3D-CRT) boost technique for locally recurrent nasopharyngeal carcinoma (NPC) in a randomised study. MATERIALS AND METHODS: Thirty-six patients with locally recurrent NPC (>6 months interval from previous radical radiotherapy, no cervical lymph-node involvement and no distant metastasis) were enrolled. Treatment included conventional external-beam radiotherapy to 54 Gy, followed by a 3D-CRT boost to the gross tumour region. Patients were randomised to three boost dose levels: 16 Gy, 20 Gy and 24 Gy for groups I, II and III, respectively, with 12 patients in each group. All boost doses were delivered in 4-Gy fractions and 3 fractions per week. Median follow-up was 27 months (range 14-44 months). RESULTS: Three-year, local-recurrence-free survival rate was significantly higher (72%) for the high dose group III than for groups I and II (37% and 28%, respectively, P = 0.047). No significant difference was found in the 3-year overall survival rate among the three groups (72%, 59% and 82% for groups I, II and III, respectively). Three year distant metastases rates were 17%, 0% and 18%, respectively. Skull-base invasion (P = 0.017) and pathology (P = 0.0006) correlated with overall survival. Treatment was well tolerated and no significant difference was observed among the three groups in acute and late toxicities (grade III toxicity is minimal: 17%, 17%, 0% of oral mucositis and 25%, 17%, 17% of nasopharyngeal mucositis in groups I, II, III, respectively, and 8% leukocytopenia only in group II; no grade IV toxicity occurred in any of the groups except for a fatal bleeding in group III). CONCLUSIONS: Re-irradiation with high-dose 3D-CRT boost technique results in high local control rate and acceptable toxicity in patients with recurrent NPC. Dose escalation to the boost volume to 78 Gy (54 Gy + 24 Gy boost) results in improved recurrence-free survival compared with lower doses. PMID- 16703747 TI - Relationship between p21 and p53 expression, human papilloma virus infection and malignant transformation in sinonasal-inverted papilloma. AB - AIMS: To identify the relationship between p21 and p53 expression, human papilloma virus (HPV) infection and malignant transformation in sinonasal inverted papilloma. MATERIAL AND METHODS: Nasal tissues, exophytic papilloma, inverted papilloma (IP) with dysplasia, IP with carcinoma and invasive squamous cell carcinoma (SCC) were stained with the monoclonal antibodies p21 and p53. In situ hybridisation for HPV DNA was also carried out for types 6/11, 16/18 and 31/33. RESULTS: Significant increased staining of p21 and p53 was observed in IP with severe dysplasia, IP with carcinoma and invasive carcinoma compared with control nasal mucosa. A significant increase of dysplasia was observed in IP in the HPV 6/11 and 16/18-positive group, compared with the HPV 6/11 and 16/18 negative group. Significant decrease in expression of p21 and p53 was observed in HPV 16/18-positive IP compared with HPV 16/18-negative IP. CONCLUSIONS: Our data raise the possibility that testing for p21, p53 and HPV may help to screen out papilloma lesions with a potential for dysplasia or carcinoma. PMID- 16703748 TI - Does the evidence support the use of concurrent chemoradiotherapy as a standard in the management of locally advanced cancer of the cervix, especially in developing countries? AB - Locally advanced cancer of the cervix (FIGO stages III and IVA) is one of the most common malignancies in developing countries. Conventional treatment has been a judicious combination of external radiotherapy and intracavitary brachytherapy. However, prompted by the results of five randomised-controlled trials (RCTs) published in close succession, The National Cancer Institute (NCI) alert in 1999, and two meta-analyses, the management of cancer of the cervix has gradually changed. Concurrent chemoradiotherapy with cisplatin alone, or in combination, is gradually being favoured for the treatment of cancer of the cervix. This overview examines whether the published evidence is sufficiently adequate to justify the use of chemoradiotherapy using cisplatin as standard care in the management of cancer of the cervix, especially in developing countries, where most women present with locally advanced cancer of the cervix. A critical review of the various phase III randomised trials and meta-analyses indicates that, although chemoradiotherapy could be a standard form of treatment for early cancer of the cervix, its role in advanced stages needs further exploration before this could be incorporated into routine clinical care. PMID- 16703749 TI - Movement of the cervix in after-loading brachytherapy: implications for designing external-beam radiotherapy boost fields. AB - AIMS: Women with invasive carcinoma of the cervix treated by chemo-radiotherapy and brachytherapy may also receive a pelvic sidewall boost using a midline shield (MLS). The purpose of this study was to assess the usefulness of implanted gold grains in detecting the movement of the cervix caused by the insertion of low dose-rate brachytherapy applicators, and its implications in designing the MLS. MATERIALS AND METHODS: The medical records of 42 women with various stages of cervical carcinoma, who were treated by radical chemo-radiotherapy, were reviewed. All of these women underwent examination under anaesthesia (EUA) and a gold-grain insertion to demarcate the vaginal tumour extent, in the antero posterior and lateral planes, before starting external-beam radiotherapy. The isocentric orthogonal films (simulator films) of external radiotherapy and brachytherapy were compared to assess the change in position of the gold grains and the consequences for the design of the MLS for parametrial and pelvic sidewall boosts. RESULTS: A significant shift in the position of the gold grains was noted in both the x (lateral) and the y (cranial/caudal) axes. The median shift of the midline, right and left lateral gold grains was 4.5, 5 and 7 mm in the x axis, whereas it was 10, 8 and 9.5 mm in the y axis, respectively. The median shift in the x and y axes was 5.5 and 9 mm, ranging from 1 to 40 mm and 1 to 45 mm, respectively. The gold grains were shifted cranially in 34 (80%) and laterally in 29 (69%) women. Thirty-two women (76.2%) received parametrial boost radiotherapy, of which 25 (59.5%) women had a customised, pear-shaped shield, and the remaining seven (16.7%) had a straight-sided, rectangular MLS. Four women (9.5%) relapsed locally, and three of them had been treated using a customised shield. In two of these four women, there was an absolute under-dosage of the central pelvis at the tip of the intra-uterine tube by 50% of the parametrial boost dose (5.4 Gy/3 fractions/3 days). CONCLUSIONS: Insertion of the gold grains during pre-treatment EUA is a useful technique in detecting the movement of the cervix. A significant shift in the position of the gold grains was noted in both the x (lateral) and the y (cranial/caudal) axes. These shifts in the cervix can result in under-dosage of the central pelvis when delivering parametrial boosts with the MLS in place on the basis of the brachytherapy check films. When designing the individualised MLS, the resulting under-dosage could be avoided by taking account of the shift in the gold grain markers and allow the delivery of a more homogenous dose to the pelvis. PMID- 16703750 TI - External-beam radiotherapy in T1-2 N0 penile carcinoma. AB - AIMS: To review the outcome of 41 patients with invasive carcinoma of the penis treated with external-beam radiotherapy using a consistent technique and dose. MATERIALS AND METHODS: Forty-one patients with carcinoma of the penis treated at Christie Hospital, Manchester, UK, between 1995 and 2000 were reviewed retrospectively. Radiotherapy was delivered using 4 MV linear accelerators with a dose of 50 Gy or 52.5 Gy in 16 fractions over 22 days. RESULTS: The distribution of patients according to stage was T1=37, T2=4, N0=40, N3=1. Median follow-up was 4.5 years. The local control rate was 62%, nodal relapse-free rate of 88%, relapse-free rate of 51% and overall survival of 88% at 5 years. All recurrences were salvaged by surgery. Penile ulceration occurred in 8% and urethral stenosis requiring dilatation in 29%. There were no penectomies for penile necrosis. CONCLUSION: EBXRT may be offered for T1-2 cancer of the penis with close surveillance to detect local recurrences early for salvage surgery without jeopardising overall survival. It remains an alternative option to penis preserving surgery and should be discussed in a multidisciplinary setting and with the patient. PMID- 16703751 TI - Prediction of urinary symptoms after 125iodine prostate brachytherapy. AB - AIMS: To evaluate the post-treatment urinary morbidity experienced by a cohort of men undergoing ultrasound-based transperineal prostate brachytherapy, as monotherapy for early stage carcinoma of the prostate. MATERIALS AND METHODS: One hundred and thirty-four consecutive patients received prostate brachytherapy between March 2000 and July 2002, and were asked to complete the International Prostate Symptom Score (IPSS) and Hospital Anxiety and Depression (HAD) questionnaires before treatment and at 1, 3, 6, 9, 12 and 18 months after implant. Data on catheterisation and surgical interventions were also recorded. Pre-treatment IPSS, dosimetry and other variables were analysed in relation to catheterisation rates and post-treatment IPSS scores at each time window. RESULTS: One hundred and eleven patients returned sufficient data for meaningful analysis. Of the patients who completed IPSS at 1 month, 85 (97%) reported deterioration in IPSS scores. This peak of symptoms, identified by a rise in median IPSS, started to improve by 3 months, and was approaching baseline by 18 months. The only significant determinants of early urinary toxicity were pre treatment IPSS, pre-treatment prostate volume and the difficulty of implant. However, prostate volume was not significant beyond 1 month. Twenty-six patients required catheterisation at a median of 10 days after implant. Significant predictors of urinary retention were pre-treatment prostate volume and pre treatment IPSS. Patients requiring catheterisation continued to have significantly higher IPSS at 18 months than patients who had never required a catheter. CONCLUSION: Brachytherapy was generally well tolerated, with urinary toxicity in most patients persisting for at least 3-6 months after prostate brachytherapy. Those whose pre-treatment prostate volume and IPSS were high experienced more severe urinary symptoms in the first few months. PMID- 16703752 TI - Plasminogen activator inhibitor-1 promoter polymorphism is not associated with the aggressiveness of disease in prostate cancer. AB - AIMS: PAI-1 (plasminogen activator inhibitors-1) regulates plasminogen activation, and is related to tumour development. This study aims to test whether the promoter polymorphism in the PAI-1 gene is related to the aggressiveness of disease in prostate cancer. MATERIALS AND METHODS: In the present study, Taqman SNP genotyping assay was used to detect PAI-1 4G/5G polymorphism in DNA from paraffin-embedded tissues of 98 Caucasian patients with prostate cancer. RESULTS: The distribution of the genotypes is in Hardy-Weinberg equilibrium. The genotype had no statistically significant relationship with other prognostic factors. Similar risks for recurrence were seen in individuals with the 4G/4G and 4G/5G genotypes compared to those with 5G/5G genotype (odds ratio [OR] 2.65, 95% CI: 0.41-16.94, P = 0.30; OR = 2.19, 95% CI: 0.38-12.49, P = 0.38). CONCLUSION: We concluded that PAI-1 promoter polymorphism is not associated with the aggressiveness of disease in prostate cancer. PMID- 16703753 TI - Pilot study of preoperative combined modality treatment for locally advanced operable oesophageal carcinoma: toxicities and long-term outcome. AB - AIMS: Paclitaxel, a radiosensitiser, has significant activity in oesophageal cancer. We aimed to conduct a feasibility study of preoperative chemoradiation using paclitaxel, cisplatin and 5-fluorouracil (5-FU). MATERIALS AND METHODS: Sixteen eligible patients were enrolled. Infusional 5-FU, paclitaxel and cisplatin were given for 6 weeks before and concurrent with radiation. Conformal radiotherapy was delivered in two phases (45 Gy in 25 fractions). RESULTS: A total of 62.5% of the patients experienced Grade 3-4 toxicities, 50% required admission; one patient died during the neo-adjuvant phase. Twelve (75%) patients had oesophagectomy, and two (12.5%) died after surgery. Pathological complete remission (PCR) and minimal residual disease were observed in 25% (95% CI 0.5 49.5%) and 18% (95% CI 0-38%) of patients, respectively, who underwent surgery. The median survival was 39.7 months (95% CI 15, not reached); 1-, 2-, 3-, and 4 year survivals were 75% (95% CI 56.5-99.5), 56.3% (36.5-86.7), 50% (30.6-81.6), and 50% (30.6-81.6), respectively. CONCLUSION: Paclitaxel, cisplatin and 5-FU (TCF)-chemoradiation is an active regimen; the current dose schedule tested is associated with unacceptable toxicity, and cannot be recommended for routine clinical use. PMID- 16703754 TI - Symptomatic responses to neoadjuvant chemotherapy for carcinoma of the oesophagus and oesophagogastric junction: are they worth measuring? AB - AIMS: Neoadjuvant chemotherapy is used to downstage locally advanced oesophagogastric cancer. This study assessed whether changes in dysphagia and weight correlated with radiological and pathological assessment of response and surgical decision-making. MATERIALS AND METHODS: All patients with locally advanced carcinoma of the lower oesophagus or oesophagogastric junction treated with neoadjuvant ECF (epirubicin, cisplatin, and 5-fluorouracil) chemotherapy from January 2000 to January 2003 were included in this study. Patients were considered to be operable depending upon their chemotherapy response. Weight and swallowing were assessed before and after chemotherapy. Statistical analysis was carried out using ANOVA, unpaired t test and Fisher's exact test. RESULTS: Seventy-eight patients (male-female ratio: 6.8: 1; median age: 62.2 years; range: 44.1-78.0 years) underwent a median of three cycles (range: 1-7) of neoadjuvant ECF chemotherapy. Forty patients (51%) gained weight, and swallowing improved in 53 patients (68%). Radiological changes (based on computed tomography) were assessed according to WHO criteria: complete response (5%), partial response (27%), stable disease (46%) and progressive disease (15%). Patients whose swallowing improved gained significantly more weight (P < 0.0001). Swallowing (P = 0.0009) was significantly improved in radiological responders but not weight (P = 0.06); when radiological non-responders were separated into stable and progressive disease, patients with progressive disease were identified as failing to gain weight (P = 0.005). Both swallowing (P < 0.0001) and weight gain (P < 0.0001) were better in patients undergoing surgery. The use of changes of weight (P = 0.42) and swallowing (P = 0.61) failed to separate pathological responders from nonresponders in the subset of patients undergoing surgery. CONCLUSIONS: Weight gain and improved swallowing are good but not absolute indicators of radiological response to chemotherapy and patient selection for surgery. However, changes in these variables are not sufficiently sensitive to identify pathological responders from non-responders. PMID- 16703755 TI - Sweat-gland tumours: a clinical review of cases in one centre over 20 years. AB - AIMS: Sweat-gland tumours (SGTs) are uncommon, but malignant varieties are very rare. We have added our data on 30 new cases seen at the Royal Marsden NHS Foundation Trust to the published literature, particularly concentrating on clinical issues. We include a literature review. MATERIALS AND METHODS: The Royal Marsden NHS Foundation Trust database was searched for cases of SGT from 1972. Data were collected on all cases, including patient demographics and tumour characteristics, treatment and outcome. RESULTS: Thirty cases were confirmed histologically to be SGTs. Fourteen were malignant, 15 benign and the degree of malignancy in one was histologically indistinguishable. Mean age was 55 years (64 for malignant, 47 for benign tumours). The 15 patients with benign tumours were almost all treated with complete excision. Those with local relapse underwent successful re-excision. Their 5-year disease-free survival was 78% and cause specific survival was 100%. Twelve of the 14 malignant tumours had localised disease at diagnosis, one had nodal disease and one had metastatic tumour nodules. All except one were treated with wide local excision. The patient with nodal involvement also had a lymph-node dissection. Two received adjuvant radiotherapy to the tumour bed. One received a melphalan limb perfusion. Eight of the 14 had no relapse. Six had locoregional relapse, and four of these also developed distant metastases. Visceral disease was always fatal. Radiotherapy and chemotherapy at relapse were unsuccessful. Five-year disease-free survival was 45%, and cause-specific survival was 57%. CONCLUSION: These rare tumours should be treated initially with complete wide local excision. In malignant tumours, lymph-node involvement is a poor prognostic sign. Wide local excision remains the primary treatment. Adjuvant radiotherapy may be useful in high-risk cases. PMID- 16703756 TI - The utility of magnetic resonance imaging in the detection of brain metastases in the staging of cutaneous melanoma. AB - AIMS: To retrospectively evaluate the use of brain magnetic resonance imaging (MRI) in the initial staging of patients with cutaneous melanoma in our melanoma unit. MATERIALS AND METHODS: The radiology request forms for brain MRI for melanoma staging for 193 consecutive patients were reviewed. Patient hospital records were also retrospectively reviewed. Patients with no histological confirmation of a cutaneous primary or patients whose scan was to primarily investigate their neurological symptoms were excluded. Records were also searched for incidental symptoms that may have been associated with brain metastases. RESULTS: One hundred patients were eligible. No patients were upstaged by MRI. Of a total of 33 patients already graded as stage IV by prior staging, 11 (33%) were found to have brain metastases. No patients graded less than stage IV were found to have brain metastases on MRI. Six out of 12 patients with incidental symptoms had metastases. Five patients graded as stage IV had asymptomatic brain metastases. CONCLUSION: We recommend brain MRI only for patients with stage IV disease and for other patients with melanoma contemplating further adjuvant therapy where brain metastases would change management. PMID- 16703757 TI - Patient set-up in radiotherapy with video-based positioning system. AB - The precision of patient position set-up is important in radiotherapy. A simple and effective scheme is proposed to calibrate the binocular cameras into the treatment machine co-ordinate system to accomplish the patient's first set-up using the video-based positioning system. We also introduce the marking of points on the surface of the body, which can be clearly imaged by computed tomography and are easily recognised from the photograph by the charge coupled device (CCD) camera. By comparing the real-time co-ordinates of the marked points with those obtained from the computed tomography, and with subsequent adjustment, the patient's first set-up in radiotherapy is realised with the video-based positioning system. PMID- 16703758 TI - Interleukin-6 in patients with aggressive and indolent non-Hodgkin's lymphoma: a predictor of prognosis? PMID- 16703759 TI - [The influence of the socio-economic structure of the breeding farms of Franches Montagnes horses on the conditions of husbandry]. AB - The socio-economic structure of the breeding farms of Franches-Montagnes horses (FM) in Switzerland is evaluated on the basis of an investigation carried out in 2002 by the Swiss FM breeding federation. Questionnaires were sent to 3500 of its members and the results include data from 968 breeding enterprises, housing a total of 3965 FM. The quality of the husbandry of FM varies according to factors such as the altitude and the geographical situation of the farms and studs. Socio economic parameters, such as the role of FM in the business, their use (breeding, driving, riding) and the age and level of professional education of the owners may also have an effect on standards of husbandry. The results show that the owners for whom FM represent a source of income more frequently keep their horses in standing stalls, but give them more time to exercise at liberty than the horses belonging to amateur breeders. Younger and better educated breeders are more likely to house their animals in groups. PMID- 16703760 TI - [Evaluation of the quality of husbandry of Franches-Montagnes horses in their breeding farms]. AB - The quality of husbandry of Franches-Montagnes horses (FM) in Switzerland is evaluated on the basis of an investigation carried out in 2002 by the Swiss FM breeding federation. Questionnaires were sent to 3500 of its members and the results include data from 968 breeding enterprises, housing a total of 3965 FM: 46.1% were breeding mares (61.0% with foal at foot), 26.5% young stock, 1.3% stallions and 26.0% non breeding stock (74.6% of which were pleasure horses and 25.4% working horses). 57.6% of the FM were housed in individual boxes with or without permanent outdoor access, 25.4% were hold in groups with or without permanent outdoor access, the remaining 17.0% were kept in standing stalls. 95.0% of the FM had at least visual contact with other equines and 99.2% had sufficient light in their stable. 88.1% were stabled on long stalk straw, while only 4.3% were bedded on other materials other than straw. The average time spent at pasture per horse and per week ranged from 96.5 +/- 51.6 hours in summer to 27.2 +/- 26.7 hours in winter. On average, a FM is used for 8.3 +/- 6.5 hours per week. Horses with an paddock at their disposal spend an average of 39.8 +/- 45.9 hours there per week. PMID- 16703762 TI - Coral disease dynamics in the central Philippines. AB - Limited quantitative research has been conducted on coral disease in the Philippines and baseline data are much needed. Field surveys for prevalence and distribution patterns were conducted from November 2002 to August 2003. Sites included the islands of Negros, Cebu, Siquijor, Panglao, Olango, Sumilon, Bantayan, Pescador, Balicassag and Palawan. In 154 belt transects, 10 026 Porites colonies were examined at 28 sites covering 3080 m2. Two syndromes, Porites ulcerative white spot (PUWS) and coral tumors, occurred at high prevalence. Tumors as high as 39.1% occurred among massive Porites, and PUWS was as high as 53.7% among massive and branching Porites. In 8 mo, 116 tagged colonies showed slow progression and low mortality. Along a 41 km human impact gradient centered on Dumaguete City (Negros), 15 sites were examined. Correlation analyses linked higher disease prevalence to anthropogenic influence (Spearman's rank correlation coefficient [r(s)] = -0.54, p = 0.04 for tumors and r(s) = -0.69, p = 0.005 for PUWS). In most sites disease prevalence was lower than in the sites near Dumaguete. High PUWS prevalence near uninhabited Sumilon Island appeared to be linked to the highly diseased reefs near Dumaguete City due to transmission of disease along a cross-shelf front formed between the Tanon Strait and Bohol Sea. Other observations included 12 potential new host species for PUWS (4 new genera and 1 octocorallia) and 5 likely new hosts for black band disease (BBD) in the Philippines, and a relatively high prevalence (7.8%) of BBD in 1 site in western Palawan. PMID- 16703761 TI - Status and progress in coral reef disease research. AB - Recent findings on the ecology, etiology and pathology of coral pathogens, host resistance mechanisms, previously unknown disease/syndromes and the global nature of coral reef diseases have increased our concern about the health and future of coral reef communities. Much of what has been discovered in the past 4 years is presented in this special issue. Among the significant findings, the role that various Vibrio species play in coral disease and health, the composition of the 'normal microbiota' of corals, and the possible role of viruses in the disease process are important additions to our knowledge. New information concerning disease resistance and vectors, variation in pathogen composition for both fungal diseases of gorgonians and black band disease across oceans, environmental effects on disease susceptibility and resistance, and temporal and spatial disease variations among different coral species is presented in a number of papers. While the Caribbean may still be the 'disease hot spot' for coral reefs, it is now clear that diseases of coral reef organisms have become a global threat to coral reefs and a major cause of reef deterioration. PMID- 16703763 TI - Coral diversity and disease in Mexico. AB - Field studies and empirical tests of the 'diversity-disease hypothesis' demonstrate the effects of species richness on disease transmission and severity in plant systems. Yet the converse, i.e. effects of disease on diversity, is rarely considered in either relatively well-studied plant systems or marine ecosystems. We investigated these effects along the Mexican Yucatan Peninsula to (1) quantify the relationship between disease prevalence and coral diversity, (2) test the hypothesis that octocoral and scleractinian disease prevalence are associated with one another, and (3) establish a long-term dataset. Aspergillosis of sea fans and 6 scleractinian diseases were documented. Prevalence of aspergillosis declined from 12.85% in 2002 to 5.26% in 2004, while prevalence of scleractinian diseases remained relatively constant at 5.7 +/- 0.8% in 2002 and 7.96 +/- 0.7% in 2004. Sites were relatively rich (71 species of octocoral and scleractinian corals) and even (E5 > 0.5). Sea fan disease prevalence was not associated with scleractinian disease prevalence, nor were there consistent associations between disease and measures of diversity. However, the most abundant octocoral and scleractinian species are susceptible to infection with several diseases, and disease may alter coral diversity in complex ways. These data represent the first in what will become a long-term dataset monitoring disease prevalence and associated changes in coral diversity. PMID- 16703764 TI - Coral diseases near Lee Stocking Island, Bahamas: patterns and potential drivers. AB - The number of coral diseases, coral species they infect, number of reported cases, and range over which these diseases are distributed have all increased dramatically in the past 3 decades, posing a serious threat to coral reef ecosystems worldwide. While some published studies provide data on the distribution of coral diseases at local and regional levels, few studies have addressed the factors that may drive these distributions. We recorded coral disease occurrence, prevalence, and severity along with temperature, sedimentation, and coral population data (species abundance and colony size) over 2 consecutive summers on reefs near Lee Stocking Island (LSI) in the Bahamas' Exuma Chain. In 2002 a total of 11092 coral colonies (all species present) were examined within a survey area of 9420 m2, and 13 973 colonies within 10 362 m2 in 2003. Similar to other reports, relatively large, framework species including Siderastrea siderea, Colpophyllia natans, and Montastraea annularis, along with the smaller Dichocoenia stokesi, were the species most susceptible to coral disease. Recurring infections were observed on individual colonies from 2002 to 2003, and were more likely for black band disease (BBD) than for either white plague (WP) or dark spots syndrome (DS). In 2002, WP and DS demonstrated clumped distributions, while BBD was randomly distributed. However, in 2003 BBD and WP were clumped. This is the first study, to our knowledge, that quantitatively documents coral disease dynamics on reefs surrounding LSI. PMID- 16703765 TI - Distribution, host range and large-scale spatial variability in black band disease prevalence on the Great Barrier Reef, Australia. AB - The prevalence and host range of black band disease (BBD) was determined from surveys of 19 reefs within the Great Barrier Reef Marine Park, Australia. Prevalence of BBD was compared among reefs distributed across large-scale cross shelf and long-shelf gradients of terrestrial or anthropogenic influence. We found that BBD was widespread throughout the Great Barrier Reef (GBR) and was present on 73.7% of the 19 reefs surveyed in 3 latitudinal sectors and 3 cross shelf positions in the summer of 2004. Although BBD occurred on all mid-shelf reefs and all but one outer-shelf reefs, overall prevalence was low, infecting on average 0.09% of sessile cnidarians and 0.1% of scleractinian corals surveyed. BBD affected approximately 7% of scleractinian taxa (25 of approximately 350 GBR hard coral species) and 1 soft coral family, although most cases of BBD were recorded on branching Acropora species. Prevalence of BBD did not correlate with distance from terrestrial influences, being highest on mid-shelf reefs and lowest on inshore reefs (absent from 66%, n = 6, of these reefs). BBD prevalence was consistently higher in all shelf positions in the northern (Cooktown/Lizard Island) sector, which is adjacent to relatively pristine catchments compared to the central (Townsville) sector, which is adjacent to a more developed catchment. BBD cases were clustered within reefs and transects, which was consistent with local dispersal of pathogens via currents, although the spread of BBD was not dependent on the density or cover of any of the coral taxa examined. In combination, these results suggest that BBD is part of the natural ecology of coral assemblages of the GBR, and its prevalence is relatively unaffected by terrestrial influences on the scales characteristic of cross-shelf gradients. PMID- 16703766 TI - Colony versus population variation in susceptibility and resistance to dark spot syndrome in the Caribbean coral Siderastrea siderea. AB - Scleractinian corals appear to be increasingly susceptible to pathogenic diseases, yet it is poorly understood why certain individuals, populations or species are more susceptible to diseases than others. Clearly an understanding of mechanisms of disease resistance in corals is essential to our understanding of patterns of disease incidence and virulence; this work must begin by examining the colony and population levels of organization. The Caribbean coral Siderastrea siderea exhibits variability in susceptibility to dark spot syndrome (DSS), a disease of unknown origin that can result in tissue necrosis. On the reef scale, variability in DSS prevalence in S. siderea occurred through time, but was not correlated with site, seawater temperature or depth. We monitored colonies of S. siderea affected by DSS, as well as their nearest neighbor controls, for 2 years in the Bahamas and found a marked decline in extent of DSS infection in October of both years. A preliminary survey of antimicrobial activity in S. siderea indicated selective activity against certain ecologically relevant bacteria. To assess whether changes in chemical defenses were responsible for the observed temporal variability in DSS prevalence, we sampled S. siderea for qualitative and quantitative analysis of chemical variability between resistant and susceptible colonies of S. siderea. These data suggest that phenotypic plasticity in antimicrobial activity may impact microbial settlement and/or survival. PMID- 16703767 TI - Consequences of yellow band disease (YBD) on Montastraea annularis (species complex) populations on remote reefs off Mona Island, Puerto Rico. AB - The rate and extent of mortality from yellow band disease (YBD) to Montastraea annularis (species complex) on reefs off Mona Island, Puerto Rico, was evaluated over 8 yr. Isolated YBD infections were first observed in 1996. Prevalence of YBD increased dramatically in 1999, with a maximum of 52 % of all M. annularis colonies infected in 1 shallow site. YBD continued to spread among adjacent, previously uninfected corals over the next 4 yr, and disease prevalence progressively increased in deeper sites. Linear rates of disease advance and tissue mortality have been slow (5 to 15 cm yr(-1)), although colonies with single YBD lesions have become infected in multiple locations. Most (85%) colonies identified with YBD in 1999 and 2000 were still affected in 2003, and these corals have lost a mean of 60% of their living tissue. Mortality from YBD is being compounded by black band disease, white plague and other syndromes; bioeroding sponges, macroalgae, cyanobacteria and other competitors have colonized tissue-denuded skeleton, minimizing the likelihood of resheeting. The deteriorating health of M. annularis is of particular concern, as these are the dominant corals on these reefs, the largest (2 to 3 m diameter and height) and presumably oldest colonies were infected with YBD more frequently than small colonies, and no recruitment has been observed. YBD is causing extensive mortality to key reef-building taxa in a remote location where anthropogenic stressors are minimal. Additional research on causes of YBD, mechanisms of infection, and strategies to mitigate YBD is needed; otherwise, M. annularis may suffer a fate similar to that of the Atlantic acroporids. PMID- 16703768 TI - Folliculinid ciliates: a new threat to Caribbean corals? AB - This is the first report of a putative pathogenic ciliate protozoan that has been associated with Caribbean corals. Previously, only 2 species of the phylum Ciliophora had been linked to coral diseases, and they were exclusive to the Indo Pacific region. In this study, a ciliate of the genus Halofolliculina was found on 10 hard coral species at the National Parks of Los Roques and Morrocoy, Venezuela. The general morphology of this ciliate is very similar to that of Halofolliculina corallasia from the Indo-Pacific, which is known to cause skeletal eroding band. None of the other 31 genera in the family Folliculinidae are known to cause diseases in corals or in any other animal species. The presence of this ciliate, which shows a prevalence comparable to that of other epizootics in the Caribbean, suggests it could be a new threat to the coral reefs of this region. PMID- 16703769 TI - Bacterial community structure associated with white band disease in the elkhorn coral Acropora palmata determined using culture-independent 16S rRNA techniques. AB - Culture-independent molecular (16S ribosomal RNA) techniques showed distinct differences in bacterial communities associated with white band disease (WBD) Type I and healthy elkhorn coral Acropora palmata. Differences were apparent at all levels, with a greater diversity present in tissues of diseased colonies. The bacterial community associated with remote, non-diseased coral was distinct from the apparently healthy tissues of infected corals several cm from the disease lesion. This demonstrates a whole-organism effect from what appears to be a localised disease lesion, an effect that has also been recently demonstrated in white plague-like disease in star coral Montastraea annularis. The pattern of bacterial community structure changes was similar to that recently demonstrated for white plague-like disease and black band disease. Some of the changes are likely to be explained by the colonisation of dead and degrading tissues by a micro-heterotroph community adapted to the decomposition of coral tissues. However, specific ribosomal types that are absent from healthy tissues appear consistently in all samples of each of the diseases. These ribotypes are closely related members of a group of alpha-proteobacteria that cause disease, notably juvenile oyster disease, in other marine organisms. It is clearly important that members of this group are isolated for challenge experiments to determine their role in the diseases. PMID- 16703770 TI - Changes in the microbial communities associated with Gorgonia ventalina during aspergillosis infection. AB - The surface mucopolysaccharide layer (SML) secreted by corals is a rich environment where bacteria live and proliferate, with population levels often being several orders of magnitude higher than in the surrounding waters (at least for culturable microbes). Some studies have suggested that these communities play an important role in energy and nutrient flux in marine environments. We hypothesize that the microbial community structure of the SML also plays a role in protection against disease. This hypothesis is based on studies that have shown differences in the bacterial composition of the mucus of healthy and diseased corals. In this study we tested the differences in the microbial communities living in association with the SML of healthy and diseased Gorgonia ventalina by comparing their metabolic profiles using Biolog EcoPlates. Overall, metabolic profiles of the coral surface microbiota were significantly different to those in the water column based on stepwise canonical discriminant analyses (CDAs). Furthermore, differences between communities living in healthy and diseased corals were also found. Changes were observed between affected and unaffected areas of the same colony, although these changes were not as obvious as between individual healthy and diseased colonies. Results suggest that the microbial communities living in the SML of G. ventalina are affected by the presence of aspergillosis, even if the area is not in direct contact with the infection. This suggests the possibility of changes in the composition of the SML throughout the colony as a response to aspergillosis infection. PMID- 16703771 TI - Longitudinal study of aspergillosis in sea fan corals. AB - Aspergillosis (a fungal disease) is affecting sea fan corals Gorgonia spp. throughout the Caribbean. To measure the impact of this disease, we established longitudinal, or in other words individual-based, monitoring studies on 3 reefs in the Florida Keys, USA, to obtain estimates of incidence, rates of disease progress, recovery, and mortality. At Western Dry Rocks (near Key West), 40 Gorgonia ventalina colonies (20 initially healthy and 20 initially diseased) were photo-monitored between June 1996 and May 1998. Additional sea fans were visually monitored during 2 localized outbreaks at Conch (May 1998 to September 1999) and Carysfort (July 2000 to May 2001) reefs located in the Upper Keys. Data from Western Dry Rocks showed that over a 2 yr period, the incidence rate was 0.58 sea fans yr(-1) and that tissue purpling can lead to tissue loss and subsequently to mortality, albeit at low frequencies. Most sea fans, once infected, maintained a low level of damage over time. Only 3 fans recovered from the disease; however 2 were subsequently re-infected. Case fatality rate was 10% (2 of 20 initially infected died), which is equivalent to 5% yr(-1). However, mortality can increase during localized outbreaks. At Conch, mortality was 46% yr(-1) among infected sea fans (compared to 8% yr(-1) at Carysfort, a less impacted site, during the same period). During an outbreak at Carysfort, mortality was 95% yr(-1) among diseased sea fans. These data clearly demonstrate the significant role aspergillosis plays in the population ecology of sea fan corals. PMID- 16703772 TI - Viruses: agents of coral disease? AB - The potential role of viruses in coral disease has only recently begun to receive attention. Here we describe our attempts to determine whether viruses are present in thermally stressed corals Pavona danai, Acropora formosa and Stylophora pistillata and zoanthids Zoanthus sp., and their zooxanthellae. Heat-shocked P. danai, A. formosa and Zoanthus sp. all produced numerous virus-like particles (VLPs) that were evident in the animal tissue, zooxanthellae and the surrounding seawater; VLPs were also seen around heat-shocked freshly isolated zooxanthellae (FIZ) from P. danai and S. pistillata. The most commonly seen VLPs were tail less, hexagonal and about 40 to 50 nm in diameter, though a diverse range of other VLP morphotypes (e.g. rounded, rod-shaped, droplet-shaped, filamentous) were also present around corals. When VLPs around heat-shocked FIZ from S. pistillata were added to non-stressed FIZ from this coral, they resulted in cell lysis, suggesting that an infectious agent was present; however, analysis with transmission electron microscopy provided no clear evidence of viral infection. The release of diverse VLPs was again apparent when flow cytometry was used to enumerate release by heat-stressed A. formosa nubbins. Our data support the infection of reef corals by viruses, though we cannot yet determine the precise origin (i.e. coral, zooxanthellae and/or surface microbes) of the VLPs seen. Furthermore, genome sequence data are required to establish the presence of viruses unequivocally. PMID- 16703773 TI - A single cyanobacterial ribotype is associated with both red and black bands on diseased corals from Palau. AB - Filamentous cyanobacteria forming red and black bands (black band disease, BBD) on 3 scleractinian corals from Palau were molecularly identified as belonging to a single ribotype. Red cyanobacterial mats sampled from infections on Pachyseris speciosa and a massive Porites sp. yielded red strains RMS1 and RMS2 respectively; the black cyanobacterial mat sampled from an infection on Montipora sp. yielded black strain BMS1. Following trials of a range of specialized media and culture conditions, 2 media, Grund and ASN-III, were identified as the best for successful isolation and culturing. Cultured cyanobacteria were examined under a light microscope to establish purity, color and morphological appearance. DNA extraction and partial sequencing of the 16S rDNA gene of both red and black cyanobacterial isolates demonstrated 100% sequence identity. These isolated strains were also found to have 99% sequence identity with an uncultured cyanobacterial strain previously identified by molecular techniques as belonging to a cyanobacterial ribotype associated with BBD-infected corals in the Caribbean. This is the first report of the successful isolation and culture of cyanobacterial strains derived from both red bands and BBD. Based on these findings, it is suggested that the classification of these 2 syndromes as separate coral diseases be postponed until further evidence is collected. PMID- 16703774 TI - Culture and identification of Desulfovibrio spp. from corals infected by black band disease on Dominican and Florida Keys reefs. AB - Black band disease (BBD) of corals is characterized as a pathogenic microbial consortium composed of a wide variety of microorganisms. Together, many of these microorganisms contribute to an active sulfur cycle that produces anoxia and high levels of sulfide adjacent to the coral surface, conditions that are lethal to coral tissue. Sulfate-reducing bacteria, as sulfide producers, are an important component of the sulfur cycle and the black band community. Previous molecular survey studies have shown multiple Desulfovibrio species present in BBD but with limited consistency between bacterial species and infections. In this study we compared 16S rRNA gene sequences of sulfate-reducing bacteria selectively cultured from 6 BBD bands on 4 coral species, Diploria clivosa, D. strigosa, D. labyrinthiformes, and Siderastrea siderea, in the Florida Keys and Dominica. The 16S rRNA gene sequences were obtained through direct sequencing of PCR products or by cloning. A BLAST search revealed that 8 out of 10 cultures sequenced were highly homologous to Desulfovibrio sp. strain TBP-1, a strain originally isolated from marine sediment. Although the remaining 2 sequences were less homologous to Desulfovibrio sp. strain TBP-1, they did not match any other sulfate-reducing (or other) species in GenBank. PMID- 16703775 TI - New roles for systemic retinoids. AB - Acitretin, the only oral retinoid indicated for the treatment of psoriasis, has shown activity when used in combination, rotational, and sequential therapy regimens with other therapies. When used with phototherapy, significantly greater activity has been observed at lower doses than with either approach as monotherapy. While initial anecdotal evidence is promising, clinical trials are needed to evaluate whether sequential or rotational use of biologic agents with acitretin may yield improved efficacy and an acceptable safety profile with decreased risk of immunosuppression. Acitretin and other retinoids also work as chemopreventative agents in psoriasis patients with extensive exposure to psoralen ultraviolet A (PUVA) and in solid-organ transplant patients where a number of studies have reported decreased numbers of squamous cell cancers when treated with acitretin. PMID- 16703776 TI - Haptens as drugs: contact allergens are powerful topical immunomodulators. AB - For the past 40 years, dermatologists have safely used contact sensitizers such as dinitrochlorobenzene (DNCB), diphenylcyclopropenone (DPCP), and squaric acid dibutylester (SADBE) for the treatment of warts, alopecia areata, and even skin cancers. Most of these studies have utilized these powerful topical immunomodulators in acetone, a volatile solvent that precludes development of contact sensitizers as products. We have overcome these problems and stabilized these topical immunomodulators in a non-volatile, nonirritating GRAS (generally regarded as safe) vehicle. The current review article covers the traditional use of contact sensitizers for a variety of benign and malignant conditions and discusses possible mechanisms in relation to developments in modem molecular immunodermatology. PMID- 16703777 TI - Topical 5% 5-fluorouracil cream in the treatment of plantar warts: a prospective, randomized, and controlled clinical study. AB - Topical 5-fluorouracil (5-FU) is an antineoplastic antimetabolite that inhibits DNA and RNA synthesis, thereby preventing cell replication and proliferation. This mechanism of action may allow topical 5-FU to be utilized in the treatment of human papilloma virus (HPV). We conducted a study comparing 5% 5-FU cream under tape occlusion versus tape occlusion alone in 40 patients presenting with plantar warts. Nineteen out of 20 patients (95%) randomized to 5% 5-FU with tape occlusion had complete eradication of all plantar warts within 12 weeks of treatment. The average time to cure occurred at 9 weeks of treatment. Three patients (15%) had a recurrence at the 6-month follow-up visit; accordingly, an 85% sustained cure rate was observed. It is concluded that use of topical 5% 5 fluorouracil cream for plantar warts is safe, efficacious, and accepted by the patient. PMID- 16703778 TI - The mechanism of lithium and beta-blocking agents in inducing and exacerbating psoriasis. AB - Documentation of psoriatic eruptions occurring with the initiation of various pharmacotherapy agents has been reported in the literature. Two such agents include lithium and beta-blocking drugs. By understanding the mechanism by which these drugs induce and exacerbate psoriasis, we may gain further understanding of the disease process of psoriasis as well as how to treat this side effect. This paper reviews the literature that has examined the mechanism by which lithium and beta-blockers may induce and exacerbate psoriasis. Mechanisms involving both immunologic and non-immunologic factors have been examined in various studies. No consensus has been reached and further investigation is needed. However, findings such as improvement with inositol supplementation in cases of lithium-induced and -exacerbated psoriasis and disparate histologic presentation of beta-blocker induced psoriasis provide suggestions that both the origin and treatment of drug induced psoriasis may be different than psoriasis that is unrelated to medications. PMID- 16703779 TI - Poly-L-lactic acid: an overview. AB - In August 2004, the US Food and Drug Administration approved a poly-L-lactic acid (PLLA)-based injectable medical device for restoration and/or correction of the signs of facial fat loss (lipoatrophy) in people with human immunodeficiency virus. As a result, the properties of the PLLA microparticles have received considerable interest from the medical community. Polylactides have a long standing history of safe use in medical applications, such as pins, plates, screws, intra-bone and soft-tissue implants, and as vectors for sustained release of bioactive compounds. The L-isomer of polylactic acid is a biodegradable, biocompatible, biologically inert, synthetic polymer. Putatively, PLLA microparticles initiate neocollagenesis as a result of a normal foreign-body reaction to their presence. The build-up of collagen over time creates volume at the site of injection, while the PLLA microparticles are metabolized to carbon dioxide and water and expelled through the respiratory system. PMID- 16703780 TI - A multicenter efficacy and tolerability evaluation of benzoyl peroxide in a 10% urea vehicle for the treatment of acne vulgaris. AB - BACKGROUND: Acne vulgaris is a common skin disease that affects 70 to 96% of individuals. Topical benzoyl peroxide has been used successfully for acne treatment; however, it may be accompanied by drying and or flaking skin. The addition of a 10% urea to the product excipient is theorized to moisturize the skin due to its humectant properties, aid in the efficacy of benzoyl peroxide due to its keratolytic properties, and effectively combat Propionibacterium acnes due to its antibacterial properties. OBJECTIVE: To assess the efficacy and tolerability of the treatment of acne vulgaris with multiple strengths of benzoyl peroxide in a 10% urea vehicle gel or cream and cleanser. METHODS: A multicenter, non-randomized, open-label study in which 1,089 patients with acne vulgaris were enrolled at 133 participating physician office sites. Qualifying and consenting patients were prescribed either 4.5% or 8.5% benzoyl peroxide in a 10% urea vehicle cream or gel and cleanser. Additional medications were permitted during the study with the exception of those containing benzoyl peroxide. The physician assessed lesion counts, both inflammatory and non-inflammatory, at baseline and Week 4. Dryness and erythema were rated by the physician on a scale from 0 (none) to 8 (severe or deep) at baseline and Week 4. RESULTS: Nine hundred sixty-three patients completed the study. The following significant treatment arms were analyzed: patients treated with 4.5%/8.5% benzoyl peroxide in a 10% urea vehicle product only, patients treated with 4.5%/8.5% benzoyl peroxide in a 10% urea vehicle products along with oral doxycycline, and patients treated with 4.5%/8.5% benzoyl peroxide in a 10% urea vehicle products along with oral minocycline. A 44% (n=567) mean reduction in total lesion count was observed after 4 weeks of treatment with 4.5%/8.5% benzoyl peroxide in a 10% urea vehicle products only. Dual therapy using oral doxycycline (n=17) proved to be even more effective with a significant mean reduction in lesion count of 52% after only 4 weeks of treatment. Dual therapy using oral minocycline (n=21) yielded a significant mean reduction in lesion count of 34% after 14 weeks of treatment. The overall tolerability of the treatment illustrated the utility of urea as a moisturizing agent. CONCLUSION: Benzoyl peroxide in a 10% urea vehicle gel or cream and cleanser, used once daily for 4 weeks was found to be both effective and well tolerated for the treatment of symptoms related to acne vulgaris. PMID- 16703781 TI - Evaluation of a nonsurgical, muscle-stimulating system to elevate soft tissues of the face and neck. AB - Non-invasive procedures for tissue tightening and elevation are new frontiers in dermasurgery. Current methods for achieving a "nonsurgical face-lift" include radiofrequency and infrared light devices which disrupt non-covalent collagen bonds and stimulate collagen production. This study evaluated a novel treatment protocol designed to achieve a nonsurgical face-lift with electrical stimulation of muscle. Thirty-five women were treated with the Pan G Lift (Pan Germinal Systems, Clearwater, Florida) to enhance skin and induce soft-tissue lifting via hypertrophy of the facial musculature. The protocol consists of biweekly treatments for 10 weeks followed by monthly maintenance treatments. Results were evaluated by both the investigating physician and patients for up to 12 months. Estimates of percentage improvement were based on a scale of 1 to 4 (1 = 0%-25% [mild]; 2 = 26%-50% [moderate]; 3 = 51%-75% [good]; and 4 = 76%-100% [marked]). After 10 weeks (20 treatments), the median lifting improvement scores as assessed by patients and physician were the following: cheeks-2.0 and 2.0, jowls-2.0 and 2.0, and texture-2.0 and 2.0, respectively. Physician-evaluated eye lift and neck lift showed similar improvement. At 12 months (32 treatments), lifting and skin texture changes (assessed by patients and physician) reached 3.0 to 4.0, respectively, while physician-evaluated eye lift and neck lift achieved at least a 3.0 at 20 and 26 treatments. At one year (32 treatments), neck lift reached a 4.0. There was no downtime and no complications occurred. The non-invasive, muscle-stimulating Pan G Lift is a novel procedure that yields significant 3 dimensional improvements of the face and neck (nonsurgical face-lift), which demonstrates progressive improvement with continued monthly treatments over time. PMID- 16703782 TI - Effective treatment of head louse with pediculicides. AB - Of the pediculicides on the market, most are not 100% ovicidal and don't have a residual activity of more than 2 days. Therefore, at least 2 treatments are necessary to control the entire louse population. In order for a pediculicide to be effective it should kill all active stages of the louse after a single treatment. Otherwise remaining lice will continue laying eggs and the following treatments will not be fully effective, at least against the eggs. However, there is no general consensus as to when the second treatment should be conducted. Taking into consideration that head louse eggs hatch between 5 to 11 days, it is suggested that a second treatment should be administered 10 days after the beginning of the treatment. This might also explain why most of the clinical trials that were conducted by treating the patients twice with an interval of 6, 7, or 8 days showed a poor efficacy, and clinical trials where the pediculicide was applied with an interval of 10 days showed an efficacy level of more than 90%. PMID- 16703783 TI - Treatment of giant keratoacanthoma with intralesional 5-fluorouracil. AB - Giant keratoacanthomas measure greater than 2 cm and have a predilection for the nose and eyelids. As a result, they often present a significant therapeutic challenge. A 36-year old woman was referred for evaluation of a 4.0 x 4.0 cm giant keratoacanthoma on the nose. The patient was treated with intralesional 5 fluorouracil, and the tumor underwent complete regression leaving a full thickness nasal cleft. The defect was then reconstructed with a paramedian forehead flap with good cosmesis. PMID- 16703784 TI - Autosensitization dermatitis associated with propolis-induced allergic contact dermatitis. AB - Propolis is a beehive product known for its anti-inflammatory properties. With its growing use, propolis-induced contact dermatitis is increasing. While the dermatitis mostly occurs on areas directly exposed to propolis, our case presented an additional eczema at a site distant from the primary propolis induced contact dermatitis twice in the same individual. We diagnosed it as an autosensitization dermatitis associated with propolis-induced allergic contact dermatitis. PMID- 16703785 TI - Local immune modulator (imiquimod 5% cream) as adjuvant treatment after incomplete Mohs micrographic surgery for large, mixed type basal cell carcinoma: a report of 3 cases. AB - BACKGROUND: Surgical excision, including Mohs micrographic surgery, is the treatment of first choice for basal cell carcinoma (BCC). Occasionally, the Mohs procedure has to be ceased prematurely for unforeseen reasons. OBJECTIVE: We sought for possibilities to adjuvantly treat patients in which superficial basal cell carcinoma remained after incomplete Mohs surgery. METHODS: We report 3 cases of patients in which Mohs micrographic surgery for their large basal cell carcinomas was ceased for different unforeseen reasons, with remaining superficial fields of BCC after the aggressive and deeply localized part had been removed. Three weeks after closure of the defect, adjuvant therapy for the superficial remnants of the tumor with imiquimod 5% cream was given for 6 weeks, covering at least 1 centimeter around the surgically treated area. RESULTS: The treatment was well accepted with no serious side effects and a good cosmetic result. No recurrences were seen after a follow-up period of 20 to 34 months. CONCLUSION: A combination of surgical intervention for the central deeper part of the BCC and topical imiquimod 5% cream for the superficial part at the border might be a therapeutic option for those patients with multiple facial BCCs, severe sun damage, and extensive surgery in the past, in whom completion of the Mohs surgical procedure for the entire tumor is not possible for different reasons. PMID- 16703786 TI - Nasal reconstruction using 20 mg/ml cross-linked hyaluronic acid. AB - According to American Society for Aesthetic Plastic Surgery (ASAPS), 200,924 people had rhinoplasties in 2005. Patients typically have surgical rhinoplasty to correct unsightly noses resulting from trauma, surgery, or heredity. Several alternatives presently exist for patients considering surgical rhinoplasty. These include injections of botulinum toxins to correct the shape of the nasal tip and the use of various fillers to correct contour and profile defects. This article presents a simple and effective alternative using hyaluronic acid for some patients requiring nasal recontouring. The procedure offers patients the opportunity to avoid the risks and expense associated with surgical correction. PMID- 16703787 TI - Facial edema induced by isotretinoin use: a case and a review of the side effects of isotretinoin. AB - Isotretinoin (13-cis-retinoic acid) is a retinoid that is used to treat cystic acne, comedonal acne, and other diseases. For the treatment of acne, isotretinoin is dosed at 0.5 to 2 mg/kg daily for 5 months with a target total dose of approximately 120 mg/kg. Its most common side effects are mucocutaneous and ocular in nature (ie, cheilitis, ocular sicca, and decreased dark adaptation). It can also cause xerosis. Patients should be made aware of these side effects before taking isotretinoin and also that utilization of moisturizers and eye drops can help to mitigate such side effects. Sometimes, however, the dose of isotretinoin needs to be decreased to reduce the induction of side effects. Isotretinoin's most significant side effect is the induction of birth defects if a fetus is exposed to isotretinoin, which is pregnancy category X. Isotretinoin should be used with 2 forms of birth control by fecund women. It can rarely increase serum levels of triglycerides, which can, if very elevated, be related to the development of pancreatitis and xanthomas. Isotretinoin's well-documented but rarer side effects include intracranial hypertension. It can induce bony changes. A review of the literature demonstrates that isotretinoin is not linked to depression and suicide. Facial swelling has been linked to isotretinoin use in 3 previous case reports. We note herein the first case of facial swelling that occurred in an acne patient being treated with isotretinoin who at the time the swelling developed had no cysts, comedones, pustules, or evidence of bacterial infection. Possible reasons for the patient's facial swelling include some type of retinoid induced angioedema, exacerbation of inflammation by isotretinoin, and isotretinoin induced capillary leak syndrome. PMID- 16703788 TI - Trachyonychia: case report and review of the literature. AB - Trachyonychia is the term used to describe nail plate roughness, pitting, and ridging that may affect 1 to 20 nails. Alopecia areata, psoriasis, lichen planus, atopic dermatitis, ichthyosis vulgaris, as well as other skin conditions have been associated with trachyonychia, but the causal relationship is often challenging to demonstrate histologically. Clinical evidence of these cutaneous disorders in conjunction with a nail matrix biopsy may help elucidate an etiology of trachyonychia, but many cases often remain idiopathic. Nail biopsy findings may match skin histology, but more commonly show spongiotic or nonspecific changes. We present an interesting case of a female with progressive development of trachyonychia in all 20 nails coinciding with a new diagnosis of sarcoidosis. PMID- 16703789 TI - The number and variety of filling agents for soft tissue augmentation are continually increasing. PMID- 16703790 TI - Principles of occlusion in implant dentistry. Interview. PMID- 16703791 TI - Management of patients after renal graft loss: an open question for nephrologists. AB - Patients undergoing renal graft failure and returning to dialysis are often regarded to like facing for the first time a substitutive treatment, without considering the technical complications, the economical impact, and the psychological implications. This review attempt, to give answers to various questions, concerning the management of vascular access, the immunosuppressive therapy, the transplantectomy, the emotional and neuropsychic aspects, and the quality of life of graft-failed patients. PMID- 16703792 TI - Plasma and erythrocyte phospholipid fatty acids composition in Serbian hemodialyzed patients. AB - Dyslipidemia is one of the possible risk factors for advanced atherosclerosis in patients with chronic renal failure. Abnormal phospholipid metabolism may play an important role in the progression of atherosclerosis in patients with renal failure. The aim of this study was to determine specific characteristics of plasma and erythrocyte phospholipid content and fatty acid composition in 37 patients with chronic renal failure on hemodialysis (HD). The results were compared with the characteristics of healthy subjects. Briefly, plasma triglyceride (p < 0.001), total cholesterol (p < 0.05), and total phospholipids (p < 0.01) levels were significantly higher and HDL-cholesterol level significantly lower (p < 0.01) in HD patients. Plasma phosphatidylcholine and phosphatidylethanolamine concentration were significantly higher (p < 0.001) in HD patients. The plasma phospholipid fatty acids composition indicated significantly (p < 0.01) higher level of oleic (18:1 n-9) and lower levels of eicopentaenoic (20:5 n-3 EPA) and docosahexaenoic (22:6 n-3 DHA) acids (p < 0.05). However, in HD patients, the relative concentration of plasma phospholipid n-6 polyunsaturated fatty acid (PUFA) was significantly lower (p < 0.05). The fatty acid composition of erythrocyte phospholipid in HD patients was modified with EPA and DHA levels significantly lowered (p < 0.05). Our results demonstrate an abnormal phospholipid metabolism and deficiency of n-3 PUFA in plasma and erythrocyte phospholipids in hemodialyzed patients. PMID- 16703793 TI - Comparison of the non-invasive methods estimating dry weight in hemodialysis patients. AB - BACKGROUND: The treatment of deranged water homeostasis of hemodialysis (HD) patients needs focusing on an accurate assessment of dry weight (DW). However, the correct estimation of post-dialysis DW is still a problem. Echocardiography of inferior caval vein diameter (ICVD) was recently considered as a reliable technique to estimate DWs of HD patients, whereas conductivity measurements and biochemical parameters remain controversial. In this study, we aimed to compare the noninvasive methods estimating DW in HD patients. METHODS: We enrolled 60 patients: 30 hypervolemic (HV) (12 M, 18 F, with a mean age of 41.9 +/- 13.6 years, mean HD duration of 38 +/- 45 months) and 30 normovolemic (NV) patients (19 M, 11 F, with a mean age of 42.2 +/- 14 years, mean HD duration of 62 +/- 51.5 months) according to clinical sign and symptoms as well as the findings on chest x-ray. Furthermore, the DWs of patients were evaluated in post-HD period in terms of echocardiography parameters [ICVD and collapse index (CI) determined by Cheriex], plasma ANP (pANP) levels (RIA), and total body water (TBW) by bioelectrical impedance (BEI). RESULTS: Forty-one of 60 patients had hypervolemic findings (68%) and 19 patients had normovolemia (32%) according to echocardiography parameters. Determination of "hypervolemia" by clinical acumen and pANP levels were not reliable, especially negative predictive values were lower as follows: sensitivity, specificity, positive predictive value, negative predictive values of clinical acumen and pANP levels: 63%, 69%, 87%, 50%, and 67%, 59%, 79%, 43%, respectively. TBW established by BEI did not correlate with ICVD and CI after HD (p > 0.05). The TBW of HV group according to echocardiography parameters was greater than NV group, but the difference was not statistically significant (27.4 +/- 6.6 kg versus 26.4 +/- 5.8 kg, respectively, p > 0.05). However, there was not any difference in the divided BSA values (1.58 +/- 0.2 kg/m2 versus 1.60 +/- 0.2 kg/m2, respectively, p > 0.05). Hypertension was seen in 37 (90%) of the echocardiographically hypervolemic patients, and the blood pressure was kept under control by previously given medication in only 7 (19%) patients. After the dry weight of the patients was corrected echocardiographically to normal limits, the blood pressure of 31 patients (86%) was normalized without antihypertensive treatment, but only in 6 patients remained the necessity of antihypertensive treatment. In addition, in 8 of 11 normotensive patients using antihypertensive drugs, assessment of their clinical and radiological findings showed normovolemia but ICVD > 11.5 mm/m2; however, the need for antihypertensive drugs disappeared when the ICVD reduced to 8-11.5/m2. CONCLUSIONS: Clinical and radiological assessment, pANP levels, and TBW established by BEI appeared to be less valuable in interpreting DW's of HD patients. In accordance with the literature, echocardiography findings have proven to be reliable, and they are important noninvasive tools that can establish an effective and rational antihypertensive treatment. PMID- 16703794 TI - Analysis of mutations in the urate transporter 1 (URAT1) gene of Japanese patients with hypouricemia in northern Japan and review of the literature. AB - BACKGROUND: Renal hypouricemia is an autosomal recessive disorder resulting from inactivating mutations in the urate transporter 1 (URAT1) encoded by SLC22A12. To date, 10 mutations have been identified and W258X in the URAT1 gene is the predominant cause in middle to southwestern Japan. However, it is still unclear whether there is a regional specific distribution of mutations in northern Japan. In this study, we analyzed mutations in the URAT1 gene of five Japanese patients with renal hypouricemia in northern Japan. METHODS: Peripheral blood mononuclear cells were isolated from patients with hypouricemia and healthy control subjects. A mutation analysis of the URAT1 gene was performed completely by direct automated sequencing of polymerase chain reaction-amplified DNA products. RESULTS: We identified two mutations. These mutations [c.269G>A (R90H) and c.774G>A (W258X)] have been reported in Japanese patients. Two of five patients were homozygotes (W258X), two carried single heterozygous mutations (W258X), and the remaining one was a compound heterozygote (R90H and W258X). CONCLUSIONS: Our study suggests that there is no regional different distribution of the URAT1 genetic mutations in Japanese with renal hypouricemia. PMID- 16703795 TI - Significance of CD25 positive cells and macrophages in noncrescentic IgA nephropathy. AB - The aim of this study was to determine whether infiltration by CD25 positive cells, macrophages, and activated macrophages in the kidney is predictive of chronic histological injury and renal prognosis in adults with noncrescentic IgA nephropathy. Renal biopsies of 36 patients with noncrescentic IgA nephropathy were examined by immunohistochemistry for glomerular and interstitial CD4, CD8, and CD25 positive cells, monocytes/macrophage (Mac387), and activated macrophages (27E10). Renal injury (glomerulosclerosis, mesangial cell hypercellularity, tubular atrophy, and interstitial fibrosis) at the time of biopsy and renal prognosis (follow-up creatinine and creatinine clearance) were assessed. The mean follow-up period was 22.5 +/- 16.5 months. The number of interstitial CD8 positive cells was the best predictor of renal injury at the time of biopsy, and was positively correlated with glomerulosclerosis (p = 0.04), tubular atrophy (p = 0.04), and interstitial fibrosis (p = 0.01) but not with mesangial cell hypercellularity. The number of interstitial Mac387 and 27E10 positive cells were the best predictors of renal prognosis (r2 = 0.33 and 0.34 respectively, both p < 0.01). These data suggest the presence of CD8 cells and macrophages in the kidney at the time of biopsy could potentially serve as pathological markers to identify patients with IgA nephropathy, which may warrant more aggressive medical therapy. PMID- 16703796 TI - Comparison of blood and peritoneal lymphocytes from continuous ambulatory peritoneal dialysis patients, asymptomatic and with peritonitis. AB - OBJECTIVE: The purpose of this study was to compare the characteristics of the blood immunophenotype of CAPD patients with and without peritonitis and to compare the phenotypes of peripheral blood lymphocytes (PBL) and peritoneal lymphocytes (PL) in CAPD patients with peritonitis. METHODS: Fifty-seven CAPD patients (20 with peritonitis and 37 without peritonitis) were recruited in the study (mean age 66,88 +/- 13,48, male/ female 16/21). Lymphocyte subsets (CD2+, CD3+, CD3+/4+, CD3+/8+, CD3-/16 + 56+, CD4/CD8 ratio) were quantitated by using monoclonal antibodies and dual-color flow cytometric analysis. With the above method we measured PBL in patients with and without peritonitis. In patients with peritonitis we also measured PL. RESULTS: CD2 were slightly decreased in patients with peritonitis. Those patients also had more intense CD3 + / CD4+ lymphopenia (p < 0.05) and larger expansion of NK cells (p < 0.05). Patients with peritonitis appeared to have a lower ratio of CD4/CD8 (p < 0.05). All the above results are shown to Table 2. Following the onset of peritonitis, a consistent finding in all patients was a significant increase in CD2 population of PL compared with PBL (85.71 +/- 9.20 versus 82.60 +/- 7.34, p < 0.05) as well as in CD3 population (77.01 +/- 13.09 versus 68.74 +/- 13.43, p < 0.05). An increased number of CD3/8 in PL compared with PBL (33.70 +/- 9.34 versus 27.98 +/- 10.77, p < 0.05) was also noted. CONCLUSIONS: In the present study, we found important immune activation in asymptomatic CAPD patients. The activation increases during peritonitis. The causes and the clinical consequences of chronic activation remain unknown. PMID- 16703797 TI - Candida parapsilosis peritonitis has more complications than other Candida peritonitis in peritoneal dialysis patients. AB - Candida parapsilosis is the most prevalent pathogen of fungal peritonitis in peritoneal dialysis (PD). The difference between C. parapsilosis peritonitis and other C. species for clinical outcomes and treatment responses to fungal peritonitis remains unclear. This retrospective study of fungal peritonitis attempts to answer that question. A total 22 patients with fungal peritonitis in 762 PD patients were enrolled in this study. The mean age of the 22 patients, 9 males and 13 females, was 54.7 +/- 12.5 years with a mean PD duration of 39.7 +/- 33.4 months. Candida species accounted for 86% (19 cases) of fungal peritonitis and 41% (9 cases) were C. parapsilosis. Thirteen (59%) patients received fluconazole as monotherapy; others received either amphotericin B alone or in combination with fluconazole. Catheters were removed for all patients. The mean duration from peritonitis onset to catheter removal was 5.8 +/- 4.1 days. Eleven (50%) patients developed severe complications, with abscess formation or persistent peritonitis after catheter removal. C. parapsilosis peritonitis had a higher complication rate than other Candida species (78% versus 20%, p = 0.012). In patients who received fluconazole as monotherapy, the rate of severe complications of C. parapsilosis peritonitis was statistically higher than those of other Candida species (100% versus 29%, p = 0.013). Because of different severity and prognosis, C. parapsilosis peritonitis in PD patients should be treated more aggressively than other Candida species. PMID- 16703798 TI - Renoprotective effect of Spirulina fusiformis on cisplatin-induced oxidative stress and renal dysfunction in rats. AB - Cisplatin is an effective chemotherapeutic agent used in the treatment of a wide array of both pediatric and adult malignancies. Dose-dependent and cumulative nephrotoxicity is the major toxicity of this compound, sometimes requiring a reduction in dose or discontinuation of treatment. Recent evidences have implicated oxidative and nitrosative stress in cisplatin-induced nephrotoxicity. Spirulina fusiformis, blue-green algae, is claimed to be a potential antioxidant. The present study was designed to explore the renoprotective potential of Spirulina fusiformis against cisplatin-induced oxidative stress and renal dysfunction. Spirulina fusiformis (500,1000,1500 mg/kg(-1) p.o.) was administered 2 days before and until 3 days after cisplatin challenge (5 mg/kg(-1) i.p.). Renal injury was assessed by measuring serum creatinine, blood urea nitrogen, creatinine and urea clearance, and serum nitrite levels. Renal oxidative stress was determined by renal TBARS levels, reduced glutathione levels, and by enzymatic activity of superoxide dismutase and catalase. A single dose of cisplatin produced marked renal oxidative and nitrosative stress and significantly deranged renal functions. Chronic Spirulina fusiformis treatment significantly and dose-dependently restored renal functions, reduced lipid peroxidation, and enhanced reduced glutathione levels, superoxide dismutase, and catalase activities. The results of the present study clearly demonstrate the pivotal role of reactive oxygen species and their relation to renal dysfunction and point to the therapeutic potential of Spirulina fusiformis in cisplatin induced nephrotoxicity. PMID- 16703799 TI - Glomerulonephritis associated with chronic inflammatory demyelinating polyneuropathy. AB - Inflammatory polyneuropathies represent an important group of neurological disorders. Review of the literature disclosed that glomerulonephritis seems to be not uncommon in acute inflammatory demyelinating polyneuropathy. On the other hand, glomerulonephritis associated with chronic inflammatory demyelinating polyneuropathy (CIDP) appears to be rare. We herein report a 60-year-old man with a clinical history, physical examination and laboratory investigations consistent with CIDP, who also had severe lower limb edema and proteinuria resistant to medical therapy. Renal biopsy showed features consistent with membranous glomerulonephritis. We review the related literature, and the immunological implications are discussed. PMID- 16703800 TI - Acute renal failure secondary to small cell lung cancer with tumor infiltration of the kidneys. AB - Acute renal failure secondary to tumor infiltration of the kidneys is uncommon and largely described in patients with lymphoma or leukemia. We report a 64-year old man previously diagnosed with limited stage small cell lung cancer who presented with acute renal failure (ARF). Renal imaging showed bilateral enlargement with features suggestive of an infiltrative process. A kidney biopsy established the diagnosis of metastatic small cell lung cancer with diffuse renal parenchymal infiltration. This case emphasizes the rare potential for cancers to metastasize to the kidneys, which can result in ARF. Early recognition of this cause of ARF is crucial, in particular, when the tumor is amenable to chemotherapy or irradiation. PMID- 16703801 TI - Interactions between serum creatinine, volume status, N-acetylcysteine, and contrast-induced nephropathy. AB - Contrast-induced nephropathy is reported to be the third leading cause of acute renal failure. The development of contrast-induced nephropathy is associated with prolonged hospitalization, the potential need for renal replacement therapy, and increased mortality. N-acetylcysteine is commonly used for the prevention of contrast-induced nephropathy despite inconsistent results from numerous clinical trials and meta-analyses. The advantage of N-acetylcysteine in most studies was based on a decrease in the serum creatinine concentration and a decrease in oxidative stress. N-acetylcysteine itself can directly lower serum creatinine concentration without improving renal function, and hypovolemia can affect the oxidative stress. In most of the N-acetylcysteine studies, there are a lack of volume status and secondary end points. The value of N-acetylcysteine for the prevention of contrast-induced nephropathy seems questionable. The interactions between serum creatinine, N-Acetylcysteine, volume status, and the development of contrast-induced nephropathy were discussed in this letter. PMID- 16703802 TI - Urine glycosyl albumin formation: relation to severe nephropathy in the poor control diabetes mellitus patients? AB - Diabetes mellitus (DM) is a frequent disorder affecting individuals of all ages. Urine glycosyl albumin is an important finding in the patients with severe diabetic nephropathy. In addition, accumulation of glycated albumin in end-stage renal failure is noted. Here, the author performed a reappraisal on the bonding energy based on quantum chemical analysis. The author calculated the bonding energy of the reaction and found that the reaction is a type of "energy providing reaction." The author hereby proposed that the nature of energy providing reaction in formation of glycosyl albumin can be an insult to the surrounding renal tissues in poorly controlled DM patients. Giving out the energy from the nearby cellular compartment during formation of glycosyl albumin might be an important pathological process leading to severe diabetic nephropathy and renal failure. PMID- 16703804 TI - Modular role set to increase. PMID- 16703803 TI - Examining electrical safety. PMID- 16703805 TI - Protective measures must be built in. AB - Recent high profile thefts from hospitals have highlighted one aspect of the overall problem of security of hospital premises. There are others aspects which NHS staff deal with on a daily basis throughout England and Wales. PMID- 16703806 TI - Study uncovers infection site risks. PMID- 16703807 TI - Web tool promotes design excellence. PMID- 16703808 TI - Effective planning key to validation. PMID- 16703809 TI - Utilities or means of life support? PMID- 16703810 TI - Air and water hygiene focus. PMID- 16703811 TI - Continuing competence for registered nurses is here to stay. PMID- 16703812 TI - Teaching new graduate nurses the tricks of the trade. PMID- 16703813 TI - Handle with care: a new curriculum for safe patient handling and movement at the University of North Carolina at Chapel Hill. PMID- 16703814 TI - Image of nursing. PMID- 16703815 TI - Familiarity with, knowledge, and utilization of standardized outcome measures among physiotherapists in Nigeria. AB - Measurement tools that assess changes in patients' clinical status are called outcome measures and their integration into clinical practice promotes objective assessment and evidence-based practice. Healthcare providers' knowledge andfamiliarity with outcome measures are pertinent to their utilization. The purpose of this descriptive survey study was to determine the levels offamiliarity, knowledge and utilization of 16 standardized outcome measures (SOM) among physiotherapists in Nigeria. A 3-part questionnaire was administered (N = 236). Part 1 elicited demographic information, Part 2 assessedfamiliarity and utilization and Part 3 assessed knowledge of the SOM. Results indicated that over 60% of the respondents had never used and were not familiar with 14 out of the 16 SOM, indicating low levels of utilization and familiarity among respondents. Respondents' knowledge about the SOM was low (3.1 +/- 2.5), and correlated positively and significantly (p < 0.05) with their levels offamiliarity with and utilization of the SOM. Working experience had a positive influence on knowledge about SOM, but influence of work setting on familiarity, utilization, and knowledge about SOM was inconclusive. This study suggests that physiotherapists in Nigeria have poor knowledge, seldom utilize, and are unfamiliar with SOM and that the use of SOM should be promoted among them. PMID- 16703816 TI - Overground and walkway ambulation with unilateral hip osteoarthritis: comparison of step length asymmetries and reproducibility of treadmill mounted force plate readings. AB - The purpose of this study was to analyze and compare the degree of step length asymmetry of patients with hip osteoarthritis during free walking and treadmill ambulation and to determine the reproducibility of treadmill based vertical ground reaction force parameters. Twelve subjects with monoarticular hip osteoarthritis undergoing total hip replacement surgery were assessed preoperatively. The assessed parameters were step length, maximal forces at heel strike and push off minimum force, force inclining and declining rate and relative stance time. The results indicated significant step length differences when comparing overground and treadmill ambulation. The extent of step length asymmetry did not have a consistent expression in different gait conditions. The study demonstrated sufficient intraday reliability for mean GRF measurements. While comparing data from treadmill and walkway locomotion of patients with osteoarthritis it should be taken into account that the amount of step length asymmetry is not identical. Patients with gait disabilities seem to have a somewhat lower level of repeatability and it appears that an increased number of practice or acclimatization trials are necessary. PMID- 16703817 TI - Diverse applications of negative pressure wound therapy: a multiple case report. AB - The purpose of this retrospective case report was to describe the results of negative pressure wound therapy (NP WT) in the treatment of wounds of various size, location and etiology in four patients. The effects of NP WT were assessed in the following patients. 1) A 68-year-old male diagnosed with cellulitis presenting with two open wounds on the left foot; 2) a 60-year-old male diagnosed with Type II diabetes mellitus presenting with a full thickness wound on the plantar surface of the right foot; 3) a 64-year-old male diagnosed with necrotizing fasciitis, diabetes mellitus, and Crohn's disease presenting with a left gluteal wound; and 4) a 47-year-old obese female diagnosed with necrotizing fasciitis presenting with a wound of the right groin and buttock. All wounds were debrided of devitalized tissue and contained nearly 100% granulation tissue prior to NPWT treatment. NPWT was applied three times per week, each application lasting 48 hrs, followed by dressing change, cleaning of the wound bed and continued debridement as required. Treatment duration, varying from 4-9 weeks, corresponded to the patient's length of stay at the sub-acute rehabilitation facility, and pressure settings ranged from 100 to 200 mmHg. Measurements of wound length, width, depth and appearance were recorded once a week. Follow-up was conducted 6-18 months post treatment to assess self-reported wound status. NPWT resulted in reductions of wound length, width, and depth, with the greatest reduction in each case being wound depth. Reduction in wound depth ranged from 75 100% (100% being complete wound closure), with four of the five wounds exceeding 90%. Reduction in wound length ranged from 32-100%, with three wounds exceeding 80%, and reduction in wound width ranged from 35-100%, with three wounds exceeding 80%. Three wounds closed completely either during therapy or soon (2-4 weeks) thereafter, and two wounds were reduced in size sufficiently for treatment via skin grafts. NPWT, applied to wounds of diverse size, location, and etiology, resulted in reductions in wound length, width, and depth in four patient cases, with the greatest influence of reducing wound depth. PMID- 16703818 TI - Use of the faces pain scale to evaluate pain of a pediatric patient with pauciarticular juvenile rheumatoid arthritis. AB - The Faces Pain Scale developed by Wong and Baker is a common assessment tool that uses cartoon-like faces to assess self reported pain in children. The purpose of this case report is to explore the appropriateness of the Faces Pain Scale as an outcome measure for a young child with pauciarticular juvenile rheumatoid arthritis. The patient was a four-year-old boy who had undergone a synovectomy on his right knee secondary to pauciarticular JRA. Each session the patient was asked to rate his pain using the Faces Pain Scale. The patient gainedfull knee range of motion and his functional mobility improved compared to initial visit. His subjective pain rating remained constant at "no hurt" throughout three weeks of visits. His functional mobility did not match his subjective rating of pain via the Faces Pain Scale. Further research is needed to determine the relationship of pain, stiffness, and function in children with rheumatic diseases. PMID- 16703819 TI - An acute care episode of a patient following bilateral total knee arthroplasty. AB - More patients with bilateral joint problems are choosing to go through one surgery to replace both joints simultaneously rather than having two separate surgeries. The goals of physical therapy following the surgery are to decrease pain, maximize range of motion (ROM) and strength, improve ambulation, and improve overall function. Few studies have focused on the importance of the acute phase of rehabilitation and most concentrate instead on long-term outcomes and outpatient care. The patient was a 62-year-old male diagnosed with osteoarthritis of both knees. The day before the initial physical therapy evaluation, the patient underwent a bilateral total knee arthroplasty. By the end of his stay, active knee ROM increased to -6 degrees to 88 degrees on the right and -6 degrees to 83 degrees on the left. Passive ROM also increased to -5 degrees to 90 degrees on the right and -5 degrees to 86 degrees on the left. The patient was able to perform supine to sit transfers with supervision and sit to stand transfers with contact guard assistance and his ambulation progressed to wvalking 100 feet on level surfaces with a rolling walker and supervision. The positive gains of this patient suggest the potential value of high motivation combined with early aggressive physical therapy. PMID- 16703820 TI - The net widens for medseekers. PMID- 16703821 TI - Automating your lab? Never fear. PMID- 16703822 TI - Derivatives. Rate swapping. PMID- 16703823 TI - Regulation. Not-so-safe skies. PMID- 16703824 TI - Technology. Tracking drugs. PMID- 16703825 TI - Regulation. N.Y. takes stock. PMID- 16703826 TI - Technology. So that's where it is. PMID- 16703827 TI - Management. Goodbye to gifts. PMID- 16703828 TI - Data page. Patient rooms come of age. PMID- 16703829 TI - The Baptist Shuffle. It's a floor cleaner, a dance move and, mostly, a state of mind. PMID- 16703830 TI - States step up. AB - In the absence of an ambitious federal effort to reduce the number of Americans without health care insurance-45 million and climbing-states around the country are considering their own solutions. The details and scope of these proposals vary widely, and in all cases, the implications for hospitals are enormous. PMID- 16703831 TI - Clinical management. As the miracles get bigger, so do the challenges in pediatrics. AB - Breakthroughs in treatment and technology are boosting outcomes for young patients, but proposed cuts in federal financing and other issues loom for children's hospitals. PMID- 16703832 TI - The diversity factor. Is your hospital doing all it can do to address its various patient populations? AB - What are the barriers to culturally competent health care? How can you collect data to help overcome disparities? How do four hospitals set the standard for meeting this imperative? PMID- 16703833 TI - Overcoming barriers to quality. An American Hospital Association special roundtable discussion. PMID- 16703834 TI - Nurturing your nurse managers. AB - Some people call it the hardest job in health care, and hospitals are pulling out all the stops to make sure they choose the best nurse managers and then help them thrive. PMID- 16703835 TI - 2005 Foster G. McGaw Prize. Winner: Venice (Calif.) family clinic. AB - Venice (Calif.) Family Clinic, the nation's largest free clinic, is the winner of this year's dollar 100,000 Foster G. McGaw Prize for excellence in community service. PMID- 16703836 TI - Using annual bed statistics to measure hospitals' surge capacity is misleading, Rutgers researchers report. PMID- 16703837 TI - Should heart attack patients go to nearest hospital or one with most advanced care? PMID- 16703838 TI - With minimal federal help, community safety nets are straining to care for uninsured immigrants. PMID- 16703839 TI - The quality army. The AHA Quality Center will accelerate the quality improvement process. PMID- 16703840 TI - Outsourcing in public hospitals: a Greek perspective. AB - PURPOSE: The purpose of this research is to investigate the extent of outsourcing, the decision-making process, the impact of outsourcing, and the future trend of outsourcing in public hospitals in Greece. DESIGN/METHODOLOGY/APPROACH: A survey instrument was designed and mailed to a random sample of 100 public hospitals in Greece and 43 usable questionnaires were received, representing a response rate of 43 percent. The survey instrument focused on the extent to which public hospitals outsource services, the decision making process for choosing an external service provider, the impact of outsourcing, and the future trend of outsourcing in public healthcare organisations. FINDINGS: Public hospitals in Greece outsource a variety of activities. Cost savings and customer satisfaction are the main factors affecting the outsourcing decision. The cooperation with a contract service provider has led to significant improvement in service quality levels. Most users are satisfied with the performance of these companies and believe that there will be an increase in the usage of these services in the future. PRACTICAL IMPLICATIONS: It provides a decision-making framework regarding outsourcing in public healthcare organisations. ORIGINALITY/VALUE: This research fills the gap in the area of outsourcing in public hospitals in Greece. PMID- 16703841 TI - The motivation to care: application and extension of motivation theory to professional nursing work. AB - PURPOSE: The purpose of this research is to describe a model of nurses' work motivation relevant to the human caring stance of professional nursing work. DESIGN/METHODOLOGY/APPROACH: The model was derived from selected theories of behavioral motivation and work motivation. Evidence-based theory addressing nurses' work motivation and nurses' motivational states and traits in relation to characteristics of organizational culture and patient health outcomes is suggested in an effort to make a distinct contribution to health services research. An integrated review of selected theories of motivation is presented, including conceptual analyses, theory-building techniques, and the evidence supporting the theoretical propositions and linkages among variables intrinsic to nurses' work motivation. FINDINGS: The model of the Motivation to Care for Professional Nursing Work is a framework intended for empirical testing and theory building. The model proposes specific leadership and management strategies to support a culture of motivational caring and competence in health care organizations. ORIGINALITY/VALUE: Attention to motivation theory and research provides insights and suggests relationships among nurses' motivation to care, motivational states and traits, individual differences that influence nurses' work motivation, and the special effects of nurses' work motivation on patient care outcomes. Suggestions for nursing administrative direction and research are proposed. PMID- 16703842 TI - Leadership and emotions in health care organizations. AB - PURPOSE: The purpose of this research is to describe the relationship between effective leadership and the leader's own ability to manage his/her emotional state. DESIGN/METHODOLOGY/APPROACH: Discusses the connection between leadership and emotions. Differentiates between management and leadership. FINDINGS: Research findings and conclusions remain in dispute. Types of behavior have been found to be integral to leadership performance. ORIGINALITY/VALUE: The paper has proposed a model for defining and differentiating between leading and managing. Five factors were found to be important to the exercise of leadership. PMID- 16703843 TI - Decentralising the NHS: rhetoric, reality and paradox. AB - PURPOSE: The purpose of this research is to examine the issues of decentralisation in the NHS. DESIGN/METHODOLOGY/APPROACH: Evidence was gathered from an empirical study in order to illustrate the effect of policy reform on relationships at both the central and the local level. In this paper issues were examined in the context of two reforms at the heart of the decentralisation agenda in the English NHS: earned autonomy and the introduction of foundation hospitals. FINDINGS: Past and current policy in the NHS reflects elements of both centralisation and decentralisation. The tension created by such opposing forces has an impact not only on inter-organisational partnership working but also on the balance of power within local health economies. ORIGINALITY/VALUE: Attempts to control the NHS exhibit unresolved contradictions and tensions. This research showed that the challenge for policymakers, managers and clinicians is to manage such tensions. PMID- 16703844 TI - Filling the assessment gap: using a learning portfolio in international development courses. AB - PURPOSE: The purpose of this research is to describe an action research project that proposed, monitored and evaluated the introduction of a learning portfolio used to replace examinations that were formerly used to assess the core courses. DESIGN/METHODOLOGY/APPROACH: An action research project was undertaken to find out whether the introduction of a portfolio was successful and what could be improved in the process of its implementation. FINDINGS: The findings indicate that portfolios are effective to support and assess the academic development of international students. ORIGINALITY/VALUE: The introduction of a portfolio to replace written examinations in the NCIHD was welcomed by all concerned. PMID- 16703845 TI - The emerging role of workplace preparedness for disaster and terrorism. PMID- 16703846 TI - Establishing evidence-informed core intervention competencies in psychological first aid for public health personnel. AB - A full-scale public health response to disasters must attend to both the physical and mental health needs of affected communities. Public health preparedness efforts can be greatly expanded to address the latter set of needs, particularly in light of the high ratio of psychological to physical casualties that often rapidly overwhelms existing mental health response resources in a large-scale emergency. Psychological first aid--the provision of basic psychological care in the short term aftermath of a traumatic event--is a mental health response skill set that public health personnel can readily acquire with proper training. The application of psychological first aid by public health workers can significantly augment front-line community-based mental health responses during the crisis phase of an event. To help achieve this augmented response, we have developed a set of psychological first aid intervention competencies for public health personnel. These competencies, empirically grounded and based on best practice models and consensus statements from leading mental health organizations, represent a necessary step for developing a public health workforce that can better respond to the psychological needs of impacted populations in disasters. PMID- 16703847 TI - Principles and practical procedures for acute psychological first aid training for personnel without mental health experience. AB - Most authorities agree that mass disasters leave in their wake a need for some form of acute mental health services. However, a review of current literature on crisis intervention and disaster mental health reveals differing points of view on the methods that should be employed (Raphael, 1986; NIMH, 2002). Nevertheless, there appears to be virtual universal endorsement, by relevant authorities, of the value of acute "psychological first aid" (American Psychiatric Association, 1954; USDHHS, 2004; Raphael, 1986; NIMH, 2002; Institute of Medicine, 2003; WHO, 2003; DoD/VAPTSD, 2004; Ritchie, et al., 2004; Friedman, Hamblin, Foa, & Charney, 2004). Psychological first aid (PFA), as an acute mental health intervention, seems uniquely applicable to public health settings, the workplace, the military, mass disaster venues, and even the demands of more well circumscribed critical incidents, e.g., dealing with the psychological aftermath of accidents, robberies, suicide, homicide, or community violence. In this document, we shall introduce the notion of psychological first aid (PFA) as one aspect of a psychological continuum of care, offer a rudimentary definition of PFA, and provide the reader with a practicalframework for its implementation utilizing the individual psychological first aid (iPFA)format. The goal of this paper is to better prepare public health, public safety, and other disaster response personnel who do not possess formal clinical mental health degrees or specialized training to provide iPFA services to primary and secondary disaster victims. PMID- 16703848 TI - Expanding disaster mental health response: a conceptual training framework for public health professionals. AB - The available research literature suggests that in disasters, individuals presenting acutely with psychologically-related complaints tend to outnumber those presenting with physical symptoms directly stemming from the injury-causing agent or event. This acute "mental health surge" can rapidly overwhelm existing community mental health resources, especially in the context of terrorism. Training professionals from outside the traditional mental health workforce in basic psychological crisis intervention may promote more efficient use of mental health services through a gatekeeper process of early intervention and appropriate referrals to mental health specialists. With their experience in patient and client services at the community level, public health professionals represent a cohort well-suited for training in and delivery of acute mental health services in disasters. In this paper, we outline a conceptual model and rationale for training public health professionals in basic crisis-oriented mental health functions (psychological first aid) in order to augment community based mental health services for affected populations in a disaster. PMID- 16703849 TI - Managing America's schools in an age of terrorism, war, and civil unrest. AB - When terrorism and natural disasters strike it is extremely important to be able to effectively manage America's schools. From the crises of Columbine, to Red Lake Minnesota, the World Trade Center and Katrina, we are reminded that schools are not immune from such disasters. It is essential that schools and communities review and augment their safe school plans and partnerships so that they can respond effectively in times of crisis. It is critically important to assess local preparedness, to update, train and practice crisis response. School officials should have specific plans in place that focus on crisis prevention, crisis preparation, crisis response and crisis recovery. This article addresses those key concerns that every school system should place on their educational agenda. Additional resources and strategies are also suggested that will promote safe and welcoming schools for all ofAmerica's children. PMID- 16703850 TI - A mental health clinician's guide to death notification. AB - The purpose of this article is to present an overview of past and present research to promote further understanding of the dynamics and process of death notification for mental health practitioners. This article includes information for interventionists to gain a better understanding of how their primary professional role and their own emotional vulnerabilities can impact the process. Additionally, the stress imposed on the interventionist, expected survivor reactions, and traumatic loss are discussed. Lastly, to help prepare mental health professionals who may be asked to assist in the process, a protocol for the systematic delivery of death notification information is suggested. PMID- 16703851 TI - Is there a duty for private employers to provide emergency mental health care services? AB - This article presents a discussion of whether employers in private companies have a duty to provide an emergency action plan with a mental health component for its employees. It discusses basic negligence concepts and focuses mainly on the "duty of care" component of negligence. It then applies the negligence concepts to private employers and discusses how private companies arguably might have a duty under the laws of negligence to provide employees with an emergency action plan, specifically a plan including mental health provisions. PMID- 16703852 TI - To err is human: supporting the patient care provider in the aftermath of an unanticipated adverse clinical outcome. AB - This article will detail how the Kaiser Permanente Healthcare Organization responded to the 2000 Institute of Medicine's report which cited statistics regarding patient safety and the cost and consequences of medical errors. It will then reveal the important role Kaiser Permanente's Employee and Physician Assistance Program serves in support of Kaiser's patient care providers who are working to reduce the medical errors that lead to adverse outcomes. While striving to increase patient safety, Kaiser Permanente's Responsible Reporting and Accountability Policy will be reviewed, with a primary focus on the components of the policy which describes how the Employee and Physician Assistance Program utilizes a model of support intended to maintain and restore the functioning and well-being of Kaiser Permanente's patient care providers. PMID- 16703853 TI - The unified approach of management and critical incident stress management: helping flight attendants and pilots in the aftermath of September 11. AB - Experience suggests that effective and appropriate responses of an organization's management after a traumatic incident can help mitigate the reactions of primary, secondary, and tertiary victims. This commentary addresses the managerial response of Southwest Airlines to the trauma induced by September 11. It highlights the effectiveness of the unity between the company, the unions, and the Critical Incident Response Team, as well as the types of interventions that were highly regarded by flight attendants and pilots who received such services. This commentary also defines the phenomenon known as fear of flight. PMID- 16703854 TI - The polymerase chain reaction and its application to clinical plastic surgery. AB - Molecular biology has become an essential component in many fields of modern medical research, including plastic surgery. Research into the molecular mechanisms underlying many disease processes offer increased understanding of the pathogenesis of disease and provide exciting therapeutic possibilities. Yet for many clinicians, the presentation of much research into molecular biological processes is couched in confusing terminology and based on scientific techniques, the basis of which are frequently difficult for the clinician to understand. The purpose of this review is to present an introduction to some of the molecular biological techniques currently in use, namely the polymerase chain reaction (PCR) and explore its applications to different aspects of plastic surgery. This review explores the role PCR now plays in all aspects of modern plastic surgery practise, with particular emphasis on normal and abnormal wound healing, the diagnosis of craniofacial anomalies, the diagnosis and treatment of cancer including melanoma and squamous cell carcinoma of the head and neck, and burns. PMID- 16703855 TI - Review of 346 patients with free-flap reconstruction following head and neck surgery for neoplasm. AB - We present our 14-year experience of free tissue transfer following surgery for head and neck neoplasm. We evaluated 346 patients mean age 57 years, 65% had squamous cell carcinoma; the most frequent sites were oral cavity and mandible (168) craniomaxillo facial region (94) pharynx +/- cervical oesophagus and oropharyngostomes (84). In 327 (95%) cases the reconstruction was a success. Flap revision was necessary in 29 (8.4% of total) and recovery was successful in 10/29. Nine patients (2.6%) died perioperatively. Poor preoperative condition, previous treatment, and requirement for vein graft were significantly associated with increased risk of major complications after surgery. Cosmetic and functional outcomes were assessed on 1-10 scales: 69 and 77% of patients, respectively, had cosmetic and functional results in the 7-10 range, indicating successful outcome. Overall survival probabilities, estimated on 338 patients with malignant disease, were 53% at 2 years and 32% at 5 years. Most patients, but not all, had advanced disease stage, and 188 (54%) had recurrent disease. Hence, overall survival rates are acceptable and justify the use of complex reconstruction procedures in such patients. PMID- 16703856 TI - Free tensor fasciae latae musculofasciocutaneous flap in reconstructive surgery: a series of 85 cases. AB - The use of tensor fasciae latae was first described as a rotation or island flap and evolved into a free flap in the late 1970s. This series of 85 patients undergoing free tensor fasciae latae transfer includes complex head and neck, abdominal wall and lower limb reconstruction. The overall success rate was 93% (79 patients), partial flap loss, 5% (four cases), and flap failure, 2% (two patients). Twelve patients (14%) required unplanned return to theatre for exploration resulting in a 75% salvage rate. We believe this series demonstrates the great versatility of this flap and highlights particular indications for its use. PMID- 16703857 TI - Modified tensor fascia lata musculofasciocutaneous flap for the coverage of trochanteric pressure sores. AB - The method most frequently used for the coverage of trochanteric pressure sores is the tensor fascia lata (TFL) flap. The authors introduce a new, hatchet-shaped incision strategy for the TFL flap, which preserves the safe blood supply of the flap and keeps the flap mobile enough. The part of the flap including the muscle is adapted to the greater trochanter. This provides a good aesthetic result without dog-ear formation at the rotation point of the flap. The donor site is closed in a V-Y fashion, and the closure does not require any skin grafting or designing a local flap. The TFL hatchet flap was used nine times on eight patients to cover trochanteric pressure sores. With one exception all patients healed. No recurrence was observed during the follow-up period, and no contour difference developed on the lateral aspect of the thigh. PMID- 16703858 TI - Functional reconstruction of Achilles tendon defects combined with overlaying skin defects using a free tensor fasciae latae flap. AB - We present our experience in functional reconstruction of the Achilles tendon with large tissue defects following after trauma and infection. To cover the skin defect and to reconstruct the Achilles tendon we used the free tensor fasciae latae (TFL) flap. From 1997 to 2003 six males, ranging from 22 to 71 (average 38.6) years, underwent this reconstructive procedure. All of them had sustained a trauma with following loss of the tendon and of the overlying tissue. After initial debridements the reconstruction with a tensor fascia latae free flap was performed. To achieve a strong distal fascia lata attachment to the calcaneal bone, we developed a special method of fixation. After vertical osteotomy in the calcaneus the distal part of the fascia flap was introduced between the bone segments, which were fixed together with a spongiosa screw. For functional outcome, it was important to fix the foot in a 90 degrees position with tension on the vascularised fascia lata. The range of motion of the ankle of the reconstructed foot showed 93.7% in comparison to the normal foot. No flap failure occurred in any of the six patients. Simultaneous soft-tissue and function restoration of the foot with TFL free flap is in our opinion an optimal one-stage reconstructive procedure. PMID- 16703859 TI - The reverse posterior interosseous flap: A solution for flap necrosis based on a review of 87 cases. AB - PURPOSE: The relationship of the flap necrosis to the placement of the flaps on the forearm was outlined and a solution in avoiding flap necrosis is discussed. METHODS: The relationship of the flap necrosis to the placement of the flaps on the forearm was investigated in 87 consecutive posterior interosseous island flaps used for the reconstruction of the hand and wrist. Fifty-eight flaps were taken from the middle and proximal thirds of the forearm, the distal edges being within the middle third of the forearm in 24 and the distal edges being within the distal third of the forearm in 34. Twenty-nine flaps were harvested within the boundaries of the distal two-thirds of the forearm, the distal edge being proximal to the distal third of the forearm in 23 and the distal edge being distal to the third of the forearm in six. The Length of the flaps varied from 6.5 cm to 12 cm. The pedicle length measure 4-13 cm. The number of perforators for each flap was recorded also. RESULTS: Flaps survived complete in 78 (89.6%) patients. Six patients had superficial necrosis of the distal part of the flap (6.8%). Three flaps were totally lost and alternative coverage was used (3.8%). The flaps that ended up with partial necrosis appeared to be in related to the site it was taken from. One flap with total necrosis and one with partial ncrosis were taken within the boundaries of the proximal third of the forearm while the distal edge was proximal to the level of distal third. One flap with total necrosis and one with partial necrosis were taken from the proximal third of the forearm while their distal edge were at the limits of the distal third of the forearm. The remaining flap ending up with total necrosis was taken from the distal third of the forearm with a short pedicle. CONCLUSIONS: For the reverse posterior interosseous flap to be reliable the flap should include the septocutaneous perforators in the distal third of the forearm. To cover distant defects reliably by a flap with a long pedicle, the flap should extend up to the distal third of the forearm to include a piece of skin with numerous perforators. PMID- 16703860 TI - A new phalloplasty technique: the free anterolateral thigh flap phalloplasty. AB - After a 10-year experience with phalloplasty in female to male gender reassignment surgery and with more than one hundred cases treated, we have introduced a new technique for creation of the neo-phallus. Between 1993 and 2002, phalloplasties were performed in our department using the free radial forearm flap or the pre-expanded suprapubic flap (modified Pryor technique). The study of long-term results and complications of these cases, as well as patient requests for a new donor site, induced us to look for an alternative flap for phalloplasty. The versatility and the low donor site morbidity of anterolateral thigh flap persuaded us to use it for phalloplasty. Since March 2003, six phalloplasties with free anterolateral thigh (ALT) flap have been performed. The results have been encouraging. The shape and the consistency of the neo-phallus are suitable, the flap can be sensate and an erectile prosthesis can easily be implanted. Penile urethral reconstruction is possible in the same operative stage. Patient satisfaction is high. The anatomy and harvesting techniques of ALT flap have already been exhaustively described by several authors and only the operative technique of phalloplasty with free ALT flap, donor site management, preliminary results and complications are reported in this paper. PMID- 16703861 TI - Functional evaluation of microvascular free flaps with positron emission tomography. AB - BACKGROUND: The aim of this study was to assess blood flow (BF) of microvascular free flaps studied with positron emission tomography (PET) in patients with head and neck squamous cell cancer (HNSCC) undergoing major radical surgery 3-4 weeks after high-dose radiotherapy. METHODS: Five patients underwent resection of the HNSCC of the oral cavity followed by microvascular reconstruction with a radial forearm flap. Regional BF in oral and neck tissues was measured with PET using radiolabelled water ([15O]H2O) twice (1-2 and 12-14 days, respectively) following radical surgery. RESULTS: In the first postoperative PET study, the median BF in the cutaneous flap area was 5.1 mL/100 g/min, and in the muscle contra-lateral to the recipient site 19.9 mL/100 g/min. A low flap-to-muscle BF ratio appeared to correlate with circulatory incongruity, and thus with poorer flap success. The follow-up study on the second postoperative week supported the results of the primary PET scan. CONCLUSIONS: This pilot study suggests that PET using [15O]H2O is a feasible method to quantitatively evaluate BF of the whole free flap in patients operated on for oral PMID- 16703862 TI - Bipaddle pectoralis major myocutaneous flap in reconstructing full thickness defects of cheek: a review of 47 cases. AB - We present a series of reconstruction of 47 patients with large full thickness cheek defects secondary to cancer ablative surgery. All patients were reconstructed primarily by bipaddle pectoralis major myocutaneous (PMMC) flap. The age of patients ranged from 25 to 85 years (mean 49.5 years). All patients were male. The size of the paddle used for mucosal defect repair ranged from 5 x 3 to 9 x 7 cm and the size of the paddle used for skin cover ranged from 4 x 4 to 9 x 8 cm. The total flap size ranged from 10 x 5 to 17 x 7 cm. One patient had complete loss of flap (2.12%). Sixteen patients had minor complications all of which settled with conservative management. The follow up period varied from 1 month to 4 years. The modification adopted in bipaddling the flap was based on anatomical location of perforators to ensure good blood supply to both paddles of flap. Placing the flap horizontally with inclusion of nipple and areola increased the reach and size of available flap. We found the technique to be anatomically sound, technically easy and reliable. Precautions taken included proper assessment of reach of the paddle, placing not more than one-third of the paddle outside the muscle and securing the skin paddle to the muscle to avoid shearing of perforators during flap raising. We conclude that this technique is a useful alternative where microsurgical free tissue transfer is not possible or as a salvage procedure in selected large full thickness oral cavity lesions. However, the disadvantages of this method include loss of nipple and areola and technical difficulty in obese patients and females. PMID- 16703863 TI - An innovation in the subcutaneous island pedicle flap for cutaneous reconstruction. AB - The aim of this study was to describe an innovation of the transposition pedicle island flap for reconstruction of the medium-sized skin defects in the face, neck and hand. Twenty-seven cases of skin tumours and scars were surgically excised and reconstructed with this island flap. ALL flap survived with primary healing postoperatively. With a follow-up from 1 to 22 months, functionally and cosmetically satisfactory outcomes were achieved. This modification of transposition island flap provides a competitive repair alternative for the treatment of medium-sized skin defects. PMID- 16703864 TI - Distraction osteogenesis in the irradiated rabbit mandible. AB - PURPOSE: The potential use of distraction osteogenesis in mandibular reconstruction has been limited by its questionable efficacy in previously radiated bone. The purpose of this study was to evaluate the effects of irradiation on mandibular distraction osteogenesis (DO). MATERIALS AND METHODS: Six rabbits received a unilateral irradiation (group R) in the mandible with a total dose of 27.0 Gy in five 5.4 Gy fractions (equivalent to 50 Gy/25 fractions). The contralateral side served as control (group C). Bilateral osteotomy was made 3 months after completion of irradiation. After a 1-week latency period bone distraction was activated at a rate of 0.5 mm twice daily for 8 days, followed by a consolidation phase of 6 weeks. New bone underwent radiographic, densitometric, histological and histomorphometric analysis. RESULTS: Two rabbits were excluded from the study, one due to infection on both sides and death in another animal. After 6 weeks of consolidation, the irradiation group had no significantly lower BMD than control group. No difference was found between the percent area of new bone in both groups. New bone was more mature and organised of group C than in groups R. Larger chondroid islands were found evident in distracted bone of group R than group C. CONCLUSIONS: It seems that DO can be feasible in previously irradiated rabbit mandible. PMID- 16703865 TI - Experimental studies on hydroxyapatite powder-carboxymethyl chitin composite: injectable material for bone augmentation. AB - We developed a hydroxyapatite (HA) powder-carboxymethyl chitin composite (HA-CMC composite) that can be injected with a 14G needle by adding distilled water. We prepared Materials I (HA = 57.0 wt%) and II (HA = 40.2 wt%) and examined their biocompatibility and osteoconductivity. With a 2-mm skin stab, the material was injected on the calvarial bone of rats. The periosteum was denuded blindly in half of the cases and preserved in the other half of the cases. Simultaneously, the material was injected subdermally into the abdominal skin to examine diachronic volume alteration of the material. Our results indicated that the new materials had biocompatibility as high as that achieved with previously developed HA materials. The difference in HA concentration did not influence the osteoconductivity, but the periosteum and the soft tissue on the cranium seemed to be an obstacle to bone ingrowth. On the other hand, the volume alteration was significantly smaller in Material I than in Material II. This composite may be especially useful in facial bone augmentation because it can be injected with only a small skin stab. When used for that purpose, the periosteum of the host bone should be denuded to facilitate bone ingrowth, and Material I will be preferable to Material II in terms of the maintenance of the initial volume. PMID- 16703866 TI - Local cutaneous arachnidism: a report of three cases and their management. AB - Bites from Loxosceles spiders can cause few symptoms to gangrenous skin necrosis or even death. To date, the treatment of the violin spider bite is largely unsatisfactory. Although no specific therapy exists, it has been suggested that heparin, steroids, dapsone, experimental antivenin and/or surgical excision may be beneficial. Three rare cases of suspected spider bite and their surgical treatment are reported. On the basis of geographical area, anamnesis and clinical symptoms, we suppose Loxoceles rufescens responsible for these bites. PMID- 16703867 TI - Letter in reply to: Keratoacanthoma observed. Br J Plast Surg (2004);57:485-501. PMID- 16703868 TI - Correspondence: avoiding lower eyelid ectropion after full thickness skin grafting. PMID- 16703869 TI - Delay of post operative radiotherapy in high risk skin cancer can be associated with recurrence. PMID- 16703870 TI - An unusual cause of trismus: Ramsay Hunt syndrome. PMID- 16703871 TI - Research options for plastic surgery. PMID- 16703872 TI - Extra abdominal desmoid tumour--a long term follow up--further comment. PMID- 16703873 TI - The impact of specialized neurocritical care. PMID- 16703874 TI - Neurocritical care. PMID- 16703875 TI - Impact of a neurointensivist on outcomes in patients with head trauma treated in a neurosciences intensive care unit. AB - OBJECT: The aim of this study was to evaluate the impact of a newly appointed neurointensivist on outcomes in head-injured patients in the neurological/neurosurgical intensive care unit (NICU). METHODS: The mortality rate, length of stay (LOS), and discharge disposition of all patients with head trauma who had been admitted to a 10-bed tertiary care university hospital NICU were compared between two 19-month periods, before and after the appointment of a neurointensivist. Data regarding these patients were collected using the hospital database and the University HealthSystem Consortium (UHC) database. Samples of medical records were reviewed for Glasgow Coma Scale (GCS) score documentation. The authors analyzed data pertaining to 328 patients before and 264 after the neurointensivist's appointment. The unadjusted mean in-hospital mortality rate increased 1.1% in the after period, but this increase was significantly lower compared with the UHC-based expected increase of 8.1% in the mortality rate during the same period (p < 0.0001). The unadjusted mean mortality rate in the NICU decreased from 13.4 to 12.9% (relative mortality rate reduction 4%) and the mean NICU LOS increased from 3.1 to 3.6 days (relative NICU LOS increase 16%), both nonsignificantly. A 51% reduction in the NICU-associated mortality rate (p = 0.01), a 12% shorter hospital LOS (p = 0.026), and 57% greater odds of being discharged to home or to rehabilitation (p = 0.009) were found in the after period in multivariate models after controlling for baseline differences between the two time periods. Better documentation of the GCS score by the NICU team was also found in the after period (from 60.4 to 82%, p = 0.02). CONCLUSIONS: The institution of a neurointensivist-led team model had an independent, positive impact on patient outcomes, including a lower NICU-associated mortality rate and hospital LOS, improved disposition, and better chart documentation. PMID- 16703876 TI - Predominance of cellular edema in traumatic brain swelling in patients with severe head injuries. AB - OBJECT: The edema associated with brain swelling after traumatic brain injury (TBI) has been thought to be vasogenic in origin, but the results of previous laboratory studies by the authors have shown that a cellular form of edema is mainly responsible for brain swelling after TBI. In this study the authors used magnetic resonance (MR) imaging techniques to identify the type of edema that occurs in patients with TBI. METHODS: Diffusion-weighted MR imaging was used to evaluate the apparent diffusion coefficient (ADC) in 44 patients with TBI (Glasgow Coma Scale Score < 8) and in eight healthy volunteers. Higher ADC values have been associated with vasogenic edema, and lower ADC values with a predominantly cellular form of edema. Regional measurements of ADC in patients with focal and diffuse injury were computed. The water content of brain tissue was also assessed in absolute terms by using MR imaging to measure the percentage of water per gram of tissue. Cerebral blood flow (CBF) was measured using stable Xe-computerized tomography (CT) studies to rule out ischemia as a cause of cellular edema. The mean ADC value in the healthy volunteers was 0.82 +/- 0.05 x 10(-3) mm2/second. The ADC values in the patients with diffuse brain injury without swelling were close to the mean for the healthy volunteers. In contrast, the patients with brain swelling had increased brain water content and low ADC values (mean 0.74 +/- 0.05 x 10(-3) mm2/second). The ADC values correlated with CT classifications. In all patients with low ADC values, the CBF values were outside the range for ischemia. CONCLUSIONS: The brain swelling observed in patients with TBI appears to be predominantly cellular, as signaled by low ADC values in brain tissue with high levels of water content. PMID- 16703877 TI - Predictive value of initial computerized tomography scan, intracranial pressure, and state of autoregulation in patients with traumatic brain injury. AB - OBJECT: The authors explored the relationship between computerized tomography (CT) scan findings and intracranial pressure (ICP) measurements obtained in the first 24 hours of monitoring to identify parameters predicting outcome in patients with severe traumatic brain injury (TBI). METHODS: Intracranial pressure, mean arterial blood pressure, cerebral perfusion pressure (CPP), and pressure reactivity index were measured continuously in 126 patients with severe TBI who were admitted to a neuroscience critical care unit. Mean values in the initial 24 hours of monitoring and in the total period of monitoring were compared with types of injury categorized on the basis of the initial CT scan according to the classification of Marshall, et al., and with Glasgow Outcome Scale scores. The initial CT scan classification correlated significantly but weakly with ICP measured during the first 24 hours of monitoring (p = 0.036) but not with mean ICP over the total time of intensive care. Both midline shift and the ratio of frontal horn diameter to internal diameter correlated with ICP in the first 24 hours (p < 0.007) and with ICP over the total monitoring period (p < 0.03). Outcome score correlated with initial CT scan findings (p = 0.018), ICP over the total monitoring period (p < 0.0023), pressure reactivity over the total monitoring period (p < 0.0002), and pressure reactivity in the first 24 hours (p < 0.0001) but not with ICP in the first 24 hours. Patients with disturbed pressure reactivity in the first 24 hours after injury had a significantly higher mortality rate than patients with intact pressure reactivity (28.6% compared with 9.5%; p < 0.001). CONCLUSIONS: Patients with severe TBI who have early loss of autoregulation have a worse prognosis. Mean ICP values in patients with diffuse TBI cannot be predicted by using the Marshall CT scan classification. PMID- 16703878 TI - Contrast ultrasonographic assessment of cerebral perfusion in patients undergoing decompressive craniectomy for traumatic brain injury. AB - OBJECT: The aims of this study were to determine whether contrast-enhanced ultrasonography (CEU) could be used for noninvasive evaluation of cerebral perfusion in patients with traumatic brain injury (TBI) and to assess the effect of decompressive surgery on cerebral perfusion as measured by CEU. METHODS: Contrast-enhanced ultrasonography with intravenous administration of a microbubble contrast agent was performed in six patients with TBI undergoing decompressive craniectomy. Contrast-enhanced ultrasonography was performed through a bur hole before craniectomy and through the calvarial defect immediately after craniectomy and on postoperative Days 1 and 2. For the latter two studies, patients were placed in the recumbent position and at a 35 degrees incline to investigate changes in perfusion produced by modulation of intracranial pressure (ICP). Cerebral microvascular blood flow increased by almost threefold immediately after craniectomy, from a mean of 7.5 +/- 6.9 (standard deviation [SD]) to 20.9 +/- 11.6 (p < 0.05), and further improved on postoperative Day 1 (mean 37.1 +/- 13.9 [SD], p < 0.05, compared with postcraniectomy microvascular blood flow) without subsequent change on Day 2. The change in microvascular perfusion correlated inversely with the initial ICP (p < 0.01), indicating less recovery of flow when preoperative ICP was markedly elevated. On postoperative Days 1 and 2, head-of-bed elevation produced an increase in microvascular perfusion on CEU (mean 37 +/- 11 compared with 51 +/- 20, p < 0.05) and a small decrease in ICP (mean 16 +/- 5 mm Hg compared with 12 +/- 4 mm Hg, p < 0.05). In patients with parenchymal hematoma, CEU provided spatial information on perfusion abnormalities in the hemorrhagic core and surrounding tissues. CONCLUSIONS: Contrast-enhanced ultrasonography has potential for the intraoperative and bedside assessment of cerebral perfusion in patients with TBI. The technique may be appropriate for evaluating responses to therapies aimed at preventing secondary ischemia and for assessing regional perfusion abnormalities. PMID- 16703879 TI - Bruns syndrome caused by intraventricular neurocysticercosis treated using flexible endoscopy. AB - OBJECT: Neurocysticercosis is the most frequent cause of hydrocephalus in adults in regions where the disease is endemic, including Latin America. The prognosis for intraventricular neurocysticercosis is worse than that for the intraparenchymal form of the disease, making treatment especially important. Although active and viable intraventricular cysts produce no reaction in the host, they can cause noncommunicating hydrocephalus, whose onset is frequently abrupt. Sometimes the increasing intracranial pressure due to obstruction of the cerebral aqueduct (ball-valve mechanism) is intermittent, producing relapsing/remitting symptoms; this life-threatening phenomenon is called "Bruns syndrome." METHODS: Between 1996 and 2004, among a group of 285 patients with neurocysticercosis and Bruns syndrome caused by cysticercal cysts of the third ventricle was diagnosed in seven patients by using magnetic resonance imaging. An endoscopic procedure with a flexible cerebral endoscope was performed, intact parasitic cysts were removed, and a complete exploration was undertaken to look for more cysticercal cysts in the whole ventricular system and the subarachnoid basal cisterns. There were no deaths or complications. All seven patients were asymptomatic during a follow-up period ranging from 1 to 5 years. CONCLUSIONS: Flexible cerebral endoscopy allows one, in a minimally invasive manner, to approach the ventricular system and subarachnoid basal cisterns and to remove intraventricular neurocysticercal cysts. Flexible endoscopy is an alternative treatment for Bruns syndrome caused by neurocysticercosis of the third ventricle. PMID- 16703880 TI - Desmoid-type fibromatoses involving the brachial plexus: treatment options and assessment of c-KIT mutational status. AB - OBJECT: Desmoid-type fibromatoses are a locally invasive soft-tissue lesion that is most commonly encountered in abdominal sites. The tumor also affects head and neck areas, particularly the supraclavicular region, where it may encase and distort the brachial plexus and compromise neurovascular structures. Neurosurgeons may be called on to treat desmoid-type fibromatoses in these sites. The authors describe their experience in treating four patients with desmoid-type fibromatoses involving the brachial plexus and report the results of immunohistochemical analysis of the tumors. METHODS: Gross-total excision with nerve sparing was the first-line therapy of choice, although the surgery was challenging. Intraoperative identification of the site of tumor origin from musculoaponeurotic tissues by the neurosurgeon was necessary in two of the four cases to achieve a correct frozen section or final pathological diagnosis. Immunostaining for c-KIT (CD117) was undertaken in all cases in light of a previous report of positive CD117 immunoreactivity in abdominal desmoid-type fibromatoses. All four tumors manifested weak focal immunostaining for c-KIT. One of the patients was given adjuvant imatinib mesylate therapy, with limited success. Subsequent polymerase chain reaction testing revealed that three of the four tumors manifested a single base pair change in exon 10 of the c-KIT gene (A to C in two cases and A to G in one case). There was local recurrence in three patients, despite gross-total excision. With the combination of surgery and radiation therapy, local disease control was achieved in three of the four patients. CONCLUSIONS: This represents the first report of c-KIT sequencing in desmoid-type fibromatoses and suggests a possible biological basis for continuing to explore the use of adjuvant imatinib mesylate therapy. PMID- 16703881 TI - Surgical treatment and outcomes in 15 patients with anterior interosseous nerve entrapments and injuries. AB - OBJECT: The authors present data obtained in 15 surgically treated patients with anterior interosseous nerve (AIN) entrapments and injuries. METHODS: Fifteen patients with AIN entrapments and injuries underwent surgery between 1967 and 1997 at Louisiana State University Health Sciences Center (LSUHSC) or Stanford University Medical Center. Patient charts were reviewed retrospectively. The LSUHSC grading system was used to evaluate the function of muscles supplied by the AIN. Nontraumatic injuries included seven AIN compressions by bone or soft tissue. Traumatic injury mechanisms consisted of stretch or contusion (six patients), injection (one patient), and burn scar (one patient). Presentations included weakness in the flexor digitorum profundus (FDP) muscle to the index finger, FDP muscle to the middle finger, pronator quadratus muscle, and flexion of the distal phalanx of the thumb. Preoperative evaluations included electromyography and nerve conduction studies as well as elbow and forearm plain radiographs. On surgery, lesions in continuity involved seven compressions, four stretch or contusion injuries, and one injection injury, all of which demonstrated nerve action potentials (NAPs) and were treated with neurolysis. Among the seven compression and four stretch or contusion injury cases, six and three patients, respectively, had LSUHSC Grade 3 or better functional recoveries postoperatively. Two stretch or contusion injuries involved lesions in continuity but demonstrated negative NAPs at surgery. Thus, each was treated using a graft repair after resection of a neuroma. There was one burn scar injury, which was treated via an end-to-end suture anastomosis, leading to a functional recovery better than Grade 3. CONCLUSIONS: Fifteen AIN entrapments or injuries responded favorably to nerve release and/or repair. PMID- 16703882 TI - Surgical treatment and outcomes in 45 cases of posterior interosseous nerve entrapments and injuries. AB - OBJECT: The authors report data in 45 surgically treated posterior interosseous nerve (PIN) entrapments or injuries. METHODS: Forty-five PIN entrapments or injuries were managed surgically between 1967 and 2004 at Louisiana State University Health Sciences Center (LSUHSC) or Stanford University Medical Center. Patient charts were reviewed retrospectively. The LSUHSC grading system was used to assess PIN-innervated muscle function. Injuries were caused by nontraumatic (21 PIN entrapments and four tumors) and traumatic (nine lacerations, eight fractures, and three contusions) mechanisms. Presentations included weakness in the extensor carpi ulnaris muscle, causing compromised wrist extension and radial drift; extensor digitorum, indicis, and digiti minimi muscles with paretic finger extension; extensor pollicis brevis and longus muscles with weak thumb extension; and abductor pollicis longus muscle with rare decreased thumb abduction due to substitutions of the median nerve-innervated abductor pollicis brevis muscle and, at 90 degrees, the extensor pollicis brevis and longus muscles. Preoperative evaluations consisted of electromyography and nerve conduction studies, elbow and forearm plain x-ray films, and magnetic resonance imaging for tumor detection. At surgery, in continuity lesions were found in 21 entrapments and three fracture related and three contusion injuries; all transmitted nerve action potentials (NAPs) and were treated with neurolysis. Five fracture-related PIN injuries, one of which was a lacerating injury, were in continuity and transmitted no NAPs; graft repairs were performed in all of these cases. Among nine lacerations, three PINs appeared in continuity, although intraoperative NAPs were absent. Two of these nerves were treated with secondary end-to-end suture anastomosis repair and one with secondary graft repair. There were six transected lacerations: three were treated with primary suture anastomosis repair, two with secondary suture anastomosis, and one with graft repair. Four tumors involving the PIN were resected. Most muscles innervated by 45 PINs had LSUHSC Grade 3 or better functional outcomes. CONCLUSIONS: Forty-five PIN entrapments or injuries responded well to PIN release and/or repair. PMID- 16703883 TI - Autologous adult human Schwann cells genetically modified to provide alternative cellular transplants in peripheral nerve regeneration. AB - OBJECT: The purpose of this study was to provide genetically modified adult human Schwann cells as tools for cell transplantation in peripheral nerve repair. The application of transfected autologous Schwann cells overexpressing regeneration promoting proteins, for example, neurotrophic or growth factors, is a promising approach in the aforementioned context. To achieve an optimal clinical outcome, it is highly important to perform enrichment, genetic modification, and retransplantation of cells in a short time. METHODS: To enable the development of these autologous cellular prostheses, the authors tested the properties of adult human Schwann cells obtained from differently treated human peripheral nerve biopsy samples. The use of "cold jet," a fast and effective enrichment procedure, as well as selective, serum-free culture conditions, resulted in very pure adult human Schwann cell cultures. Using an optimized electroporation protocol, as many as 48.4% of adult human Schwann cells were successfully transfected. CONCLUSIONS: The authors present a very fast protocol to establish adult human Schwann cell cultures that demonstrably express plasmid proteins after plasmid DNA insertion by electroporation. These autologous human Schwann cells transfected to enhance the endogenous production of regeneration-supporting proteins will likely constitute a major component of tissue-engineered peripheral nerve grafts. PMID- 16703884 TI - Quantitation of and superficial surgical landmarks for the anterior interosseous nerve. AB - OBJECT: There are scant data regarding the anterior interosseous nerve (AIN) in the neurosurgical literature. In the current study the authors attempt to provide easily identifiable superficial osseous landmarks for the identification of the AIN. METHODS: The AIN in 20 upper extremities obtained in adult cadaveric specimens was dissected and quantified. Measurements were obtained between the nerve and surrounding superficial osseous landmarks. The AIN originated from the median nerve at mean distances of 5.4 cm distal to the medial epicondyle of the humerus and 21 cm proximal to the ulnar styloid process. The distance from the origin of the AIN to its branch leading to the flexor pollicis longus muscle and to the point it travels deep to the pronator quadratus (PQ) muscle measured a mean 4 and 14.4 cm, respectively. The mean distance from the AIN branch leading to the flexor pollicis longus muscle to the proximal PQ muscle was 12.1 cm, and the mean distance between this branch and the ulnar styloid process was 7.2 cm. The mean diameter of the AIN was 1.6 mm at the midforearm. CONCLUSIONS: Additional landmarks for identification of the AIN can aid the neurosurgeon in more precisely isolating this nerve and avoiding complications. Furthermore, after quantitation of this nerve, the AIN branches can be easily used for neurotization of the median and ulnar nerves, and with the aid of a transinterosseous membrane tunneling technique, passed to the posterior interosseous nerve. PMID- 16703885 TI - Surgical anatomy of the cervical and infraclavicular parts of the long thoracic nerve. AB - OBJECT: There is insufficient information in the neurosurgical literature regarding the long thoracic nerve (LTN). Many neurosurgical procedures necessitate a thorough understanding of this nerve's anatomy, for example, brachial plexus exploration/repair, passes for ventriculoperitoneal shunt placement, pleural placement of a ventriculopleural shunt, and scalenotomy. In the present study the authors seek to elucidate further the surgical anatomy of this structure. METHODS: Eighteen cadaveric sides were dissected of the LTN, anatomical relationships were observed, and measurements were obtained between it and surrounding osseous landmarks. The LTN had a mean length of 27 +/- 4.5 cm (mean +/- standard deviation) and a mean diameter of 3 +/- 2.5 mm. The distance from the angle of the mandible to the most proximal portion of the LTN was a mean of 6 +/- 1.1 cm. The distance from this proximal portion of the LTN to the carotid tubercle was a mean of 3.3 +/- 2 cm. The LTN was located a mean 2.8 cm posterior to the clavicle. In 61% of all sides the C-7 component of the LTN joined the C-5 and C-6 components of the LTN at the level of the second rib posterior to the axillary artery. In one right-sided specimen the C-5 component directly innervated the upper two digitations of the serratus anterior muscle rather than joining the C-6 and C-7 parts of this nerve. The LTN traveled posterior to the axillary vessels and trunks of the brachial plexus in all specimens. It lay between the middle and posterior scalene muscles in 56% of sides. In 11% of sides the C-5 and C-6 components of the LTN traveled through the middle scalene muscle and then combined with the C-7 contribution. In two sides, all contributions to the LTN were situated between the middle scalene muscle and brachial plexus and thus did not travel through any muscle. The C-7 contribution to the LTN was always located anterior to the middle scalene muscle. In all specimens the LTN was found within the axillary sheath superior to the clavicle. Distally, the LTN lay a mean of 15 +/- 3.4 cm lateral to the jugular notch and a mean of 22 +/- 4.2 cm lateral to the xiphoid process of the sternum. CONCLUSIONS: The neurosurgeon should have knowledge of the topography of the LTN. The results of the present study will allow the surgeon to better localize this structure superior and inferior to the clavicle and decrease morbidity following invasive procedures. PMID- 16703886 TI - Superficial surgical landmarks for identifying the posterior interosseous nerve. AB - OBJECT: There is a paucity of information in the neurosurgical literature regarding the surgical anatomy surrounding the posterior interosseous nerve (PIN). The goal of the current study was to provide easily recognizable superficial bone landmarks for identification of the PIN. METHODS: Thirty-four cadaveric upper extremities obtained from adults were subjected to dissection of the PINs, and measurements were made between this nerve and surrounding superficial bone landmarks. In all specimens the main radial trunk was found to branch into its superficial branch and PIN at the level of the lateral epicondyle of the humerus. Proximally, the PIN was best identified following dissection between the brachioradialis and extensor carpi radialis longus and brevis muscles. At its exit site from the supinator muscle, the PIN was best identified after retraction between the extensor carpi radialis longus and brevis and extensor digitorum communis muscles. This site was a mean distance of 6 cm distal to the lateral epicondyle of the humerus. No compression of the PIN by the tendon of origin of the extensor carpi radialis brevis muscle was seen. One specimen was found to have a proximally split PIN that provided a previously undefined articular branch to the elbow joint. The mean diameter of the PIN proximal to the supinator muscle was 4.5 mm. The leash of Henry crossed the PIN in all but one specimen and was found at a mean distance of 5 cm inferior to the lateral epicondyle. The PIN exited the distal edge of the supinator muscle at a mean distance of 12 cm distal to the lateral epicondyle of the humerus. Here the mean diameter of the PIN was 4 mm. The exit site from the distal edge of the supinator was found to be at a mean distance of 18 cm proximal to the styloid process of the ulna. This exit site for the PIN was best identified following dissection between the extensor carpi radialis longus and brevis and extensor digitorum communis muscles. The distal articular branch of the PIN was found to have a mean length of 13 cm and the proximal portion of this terminal segment was located at a mean distance of 7.5 cm proximal to the Lister tubercle. CONCLUSIONS: The addition of more anatomical landmarks can help the neurosurgeon to be more precise in identifying the PIN and in avoiding complications during surgery in this region. PMID- 16703887 TI - Quantitation of and landmarks for the muscular branches of the ulnar nerve to the forearm for application in peripheral nerve neurotization procedures. AB - OBJECT: In neurotization procedures, donor nerves--either whole or in part-with relatively pure motor function can be carefully chosen to provide the optimal nearby motor input with as little donor site morbidity as possible. In this context, the ulnar nerve branches to the forearm muscles are relatively dispensable; however, quantitation of and landmarks for these branches are lacking in the literature. METHODS: The ulnar branches to the flexor carpi ulnaris (FCU) and flexor digitorum profundus (FDP) muscles in 20 upper extremities obtained in adult cadaveric specimens were dissected and quantified. In the forearm, a mean of four nerve branches led to the FCU and FDP muscles. A mean of 3.4 branches led to the FCU muscle; of these, one to three were medial branches and zero to two were lateral. Medial branches to the FCU muscle originated a mean of 2.7 cm inferior to the medial epicondyle. Lateral branches to the FCU muscle originated at a mean of 3.3 cm inferior to the medial epicondyle. The mean length of the medial branches was 3.2 cm, whereas the mean length of the lateral branches was 3.3 cm. All nerves had a single trunk for the FDP muscle, and in all specimens this branch was located deep to the main ulnar nerve trunk, originating from the ulnar nerve a mean of 2.7 cm inferior to the medial epicondyle. These branches had a mean length of 5.6 cm. The mean diameter of all medial and lateral branches to the FCU muscle was 1 mm, and the mean diameter of the branch to the FDP muscle was 2.1 mm. All branches to both the FCU and FDP muscles arose from the ulnar nerve, over its first approximately 5 cm from the level of the medial epicondyle. Additionally, all branches could be easily lengthened by gentle proximal dissection from the main ulnar nerve. CONCLUSIONS: Ulnar branches to the forearm can be easily localized and used for neurotization procedures. The branch to the FDP muscle had the greatest diameter and longest length, easily reaching the median nerve and posterior interosseous nerve via a transinterosseous membrane tunneling procedure. Furthermore, this branch could be teased away from the main ulnar nerve trunk and made to reach the distal branches of the musculocutaneous nerve in the arm. PMID- 16703888 TI - Use of an intact sensory nerve to bridge a motor nerve defect: an experimental study. AB - OBJECT: End-to-side neurorrhaphy has recently became popular for peripheral nerve repair. Although this method is mainly indicated in nerve defects in which there is an absent proximal nerve stump, bridging a motor nerve defect by coapting the proximal and distal ends of the defect to a neighboring mixed nerve in an end-to side fashion has been another experimental use of this method. In this situation, however, the source of the regenerating axons is unclear because the axons in both the proximal end of the defect and the bridging intact nerve have the capacity for regeneration. The goal of this study was to identify the source of the regenerating axons. METHODS: In this experimental study, the authors used a sensory nerve to bridge a motor nerve defect so that they could elucidate the source of the regenerating motor axons in the distal part of the motor nerve. One advantage of using a sensory nerve was that it eradicated the risk of damaging another motor nerve. Tests used in the analysis included gait evaluation, electrophysiological tests, and histological assessment. CONCLUSIONS: Results of this study showed that, in the rat model, a sensory nerve can be used to bridge a motor nerve defect, thereby eliminating the need for nerve grafting. PMID- 16703889 TI - Amplitude and phase of cerebrospinal fluid pulsations: experimental studies and review of the literature. AB - OBJECT: A recently developed model of communicating hydrocephalus suggests that ventricular dilation may be related to the redistribution of pulsations in the cranium from the subarachnoid spaces (SASs) into the ventricles. Based on this model, the authors have developed a method for analyzing flow pulsatility in the brain by using the ratio of aqueductal to cervical subarachnoid stroke volume and the phase of cerebrospinal fluid (CSF) flow, which is obtained at multiple locations throughout the cranium, relative to the phase of arterial flow. METHODS: Flow data were collected in a group of 15 healthy volunteers by using a series of images acquired with cardiac-gated, phase-contrast magnetic resonance imaging. The stroke volume ratio was 5.1 +/- 1.8% (mean +/- standard deviation). The phase lag in the aqueduct was -52.5 +/-16.5 degrees and the phase lag in the prepontine cistern was -22.1 +/- 8.2 degrees. The flow phase at the level of C-2 was -5.1 +/- 10.5 degrees, which was consistent with flow synchronous with the arterial pulse. The subarachnoid phase lag ventral to the pons was shown to decrease progressively to zero at the craniocervical junction. Flow in the posterior cervical SAS preceded the anterior space flow. CONCLUSIONS: Under normal conditions, pulsatile ventricular CSF flow is a small fraction of the net pulsatile CSF flow in the cranium. A thorough review of the literature supports the view that modified intracranial compliance can lead to redistribution of pulsations and increased intraventricular pulsations. The phase of CSF flow may also reflect the local and global compliance of the brain. PMID- 16703891 TI - Intraneural perineurioma of the third cranial nerve: occurrence and identification. Case report. AB - Intraneural perineurioma is a true but rare neoplasm that originates from perineurial cells and mainly affects peripheral nerves. It must be distinguished from other hypertrophic neuropathies that are either inflammatory or demonstrate an onion-bulb formation that originates from Schwann cells. Complying with this strict definition, only three additional cases of cranium-related perineurioma have been identified: two lesions arose extracranially and involved cranial nerves, and one occurred intracranially but did not involve a nerve. The authors describe a 27-year-old woman who presented with left third cranial nerve palsy and was found to harbor a mass lesion in the superior orbital fissure and cavernous sinus. After subtotal resection had been performed, pathological studies confirmed the presence of perineurial tumor cells in a pseudo-onion bulb formation. The cells stained positively for epithelial membrane antigen but not for S100 protein, clearly distinguishing the disease from one that originates in Schwann cells. PMID- 16703890 TI - Superficial peroneal nerve syndrome: an unusual nerve entrapment. Case report. AB - Lower-extremity pain and paresthesia have multiple origins. Early recognition of the symptoms of peripheral nerve entrapment leads to timely treatment and avoids the cost of unnecessary studies. The authors report on a case of superficial peroneal nerve syndrome resulting from nerve herniation through a fascial defect, which was responsive to surgical treatment. This 22-year-old man presented with pain and paresthesias over the lateral aspect of the right calf and the dorsum of the foot without motor weakness. Exercise led to the formation of a tender bulge approximately 12 cm above the lateral malleolus. Percussion of this site worsened his symptoms. Radiography and electromyography studies were nondiagnostic. The patient underwent surgical decompression that involved division of the fascia overlying the nerve and neurolysis of the superficial peroneal nerve. The operation resulted in symptom-free relief. Superficial peroneal nerve syndrome is an entrapment neuropathy that results from mechanical compression of the nerve at or near the point where the nerve pierces the fascia to travel within the subcutaneous tissue. Surgical decompression of the mechanical entrapment usually provides relief from pain and paresthesia. PMID- 16703892 TI - Self-inflicted nail-gun injury with 12 cranial penetrations and associated cerebral trauma. Case report and review of the literature. AB - In this case report, the authors describe a 33-year-old man who presented with headache due to the presence of 12 nail-gun nails impacted in his cranium and cerebral parenchyma. The authors also review the relevant literature regarding penetrating brain injury. The patient's physical examination revealed a Glasgow Coma Scale score of 15 and impairment of abduction of the right eye and abduction of the jaw producing dysarthria; the remaining results of the neurological examination were normal. Both x-ray films and computerized tomography (CT) scans of the head revealed the presence of 12 nails, the majority of which were located intracranially. A scattering artifact limited the ability of CT scanning to demonstrate any intracranial hemorrhage. Angiography did not demonstrate any evidence of traumatic vascular injury. After general anesthesia had been induced in the patient, the nails were removed in the operating room. Following removal of the final nail, a small left temporal craniotomy was performed to control hemorrhage from a tear in the left middle meningeal artery. Despite the development of a postoperative insular hematoma, the patient was discharged home with minimal deficits. This patient is the only known survivor of the largest number of foreign objects (12) to penetrate the skull intentionally. Overall, self-inflicted nail-gun injuries are less common than accidental discharges. A review of the literature, however, suggests that for penetrating brain injury, self-infliction is the more common mechanism. For those patients who survive such an injury, clinical decision making must focus on preventing further cortical or vascular damage. A rational management strategy should permit these patients to be discharged with no additional injury. PMID- 16703893 TI - Bilateral vidian nerve schwannomas associated with facial palsy. Case report and review of the literature. AB - Intracranial schwannomas are relatively common benign tumors arising from Schwann cells. Among the cranial nerves, the vestibular division of the vestibulocochlear nerve is the site most commonly affected by these lesions, followed by the trigeminal nerve. The authors report a case of bilateral schwannomas arising from both of the pterygoid canals. A 13-year-old girl presented with intermittent headaches and left-sided facial palsy. Preoperative computerized tomography scans and magnetic resonance images revealed nonenhancing round masses within the bilateral vidian canals, bone erosion, and sclerosis. The transnasal transseptal transsphenoidal approach was used to remove the masses. Postoperatively, the patient recovered uneventfully. On histopathological examination, the masses were confirmed as schwannomas. The clinical presentation and probable histogenesis of schwannomas arising in this location are discussed together with a review of the literature. PMID- 16703894 TI - Regression of intracranial rosai-dorfman disease following corticosteroid therapy. Case report. AB - Rosai-Dorfman disease (RDD) is an idiopathic histioproliferative disorder usually presenting with massive, painless lymphadenopathy. Extranodal involvement has been reported including at least 50 cases affecting the central nervous system (CNS). The treatment of CNS RDD as reported in the literature has primarily involved a surgical technique. The authors report on the case of a 53-year-old man presenting with multiple skull base lesions mimicking meningiomas. The patient suffered visual deterioration and underwent a right orbitopterional craniotomy as well as optic nerve decompression. Histopathological analysis revealed histiocytic cells and emperipolesis consistent with RDD. Following surgery, corticosteroid agents were administered, leading to marked resolution of both the remaining surgically untreated lesions and the balance of the patient's symptoms. This report represents the first case of the resolution of intracranial RDD following corticosteroid therapy. Corticosteroid agents should be considered an effective option in the treatment of CNS RDD. PMID- 16703895 TI - Conduction aphasia as a function of the dominant posterior perisylvian cortex. Report of two cases. AB - Assessment of eloquent functions during brain mapping usually relies on testing reading, speech, and comprehension to uncover transient deficits during electrical stimulation. These tests stem from findings predicted by the Geschwind Wernicke hypothesis of receptive and expressive cortices connected by white matter tracts. Later work, however, has emphasized cortical mechanisms of language function. The authors report two cases that demonstrate that conduction aphasia is cortically mediated and can be inadequately assessed if not specifically evaluated during brain mapping. To determine the distribution of language on the dominant cortex, electrical cortical stimulation was performed in two cases by using implanted subdural electrodes during brain mapping before epilepsy surgery. A transient isolated deficit in repetition of language was reported during stimulation of the posterior portion of the dominant superior temporal gyrus in one patient and during stimulation of the supramarginal gyrus in the other patient. These cases demonstrate a localization of language repetition to the posterior perisylvian cortex. Brain mapping of this region should include assessment of verbal repetition to avoid potential deficits resembling conduction aphasia. PMID- 16703896 TI - Persistent intracranial hypertension caused by superior sagittal sinus stenosis following depressed skull fracture. Case report and review of the literature. AB - Intracranial hypertension caused by a compound depressed skull fracture on the posterior part of the superior sagittal sinus is a rare condition, and nonspecific symptoms and signs can delay appropriate diagnosis and treatment. The authors report on a case of intracranial hypertension that persisted despite conservative treatment, including anticoagulation therapy, which did not improve severe flow disturbance related to the venous sinus compression. Management of this rare condition is discussed and the literature is reviewed. PMID- 16703897 TI - Penetrating screwdriver injury to the brainstem. Case illustration. PMID- 16703898 TI - Subtemporal transtentorial approach. PMID- 16703899 TI - Subtemporal trantentorial approach. PMID- 16703900 TI - Infratentorial hygroma. PMID- 16703901 TI - Comparison of operative and nonoperative treatment for thoracolumbar burst fractures in patients without neurological deficit: a systematic review. AB - OBJECT: Despite extensive published research on thoracolumbar burst fractures, controversy still surrounds which is the most appropriate treatment. The objective of this study was to evaluate the scientific literature on operative and nonoperative treatment of patients with thoracolumbar burst fractures and no neurological deficit. METHODS: In their search of the literature, the authors identified all possible relevant studies concerning thoracolumbar burst fracture without neurological deficit. Two independent observers performed study selection, methodological quality assessment, and data extraction in a blinded and objective manner for all papers identified during the search. In a synthesis of the literature, the authors obtained evidence for both operative and nonoperative treatments. CONCLUSIONS: There is a lack of evidence demonstrating the superiority of one approach over the other as measured using generic and disease-specific health-related quality of life scales. There is no scientific evidence linking posttraumatic kyphosis to clinical outcomes. The authors found that there is a strong need for improved clinical research methodology to be applied to this patient population. PMID- 16703902 TI - Chimney sublaminar decompression for degenerative lumbar spinal stenosis. AB - OBJECT: The authors evaluated the efficacy and safety of so-called chimney sublaminar decompression, a new technique to decompress the degenerative stenotic lumbar spinal canal without stripping of the paravertebral muscles. METHODS: Eighteen patients (nine men and nine women whose mean age was 67 years) with symptoms of claudication were selected to undergo chimney sublaminar decompression. The duration of symptoms was greater than 6 months in 17 patients. Two lumbar segments were involved in seven patients, three in eight, and four in the remaining three patients. Central canal stenosis was present in 13 patients, and lateral recess stenosis in five patients. Mild spondylolisthesis was noted in seven patients. All the patients underwent chimney sublaminar decompression. After surgery, mild wound pain developed in 14 patients, moderate wound pain in two, and severe wound pain in two. The postoperative hospital stay was 4 days or fewer in 14 patients. At follow-up examination, excellent, good, and fair outcomes were achieved in 11, five, and two patients, respectively. No patient required a body brace, and no worsening of preexisting spondylolisthesis was detected. The spinal canal was increased to two- to 6.8-fold (mean 4.2-fold) the preoperative size. CONCLUSIONS: Compared with laminectomy or endoscopic surgery, the aforementioned chimney sublaminar decompression technique was an equally effective and less invasive technique in the treatment of degenerative lumbar canal stenosis. PMID- 16703903 TI - Cervical radiofrequency neurotomy in patients with chronic whiplash: a study of multiple outcome measures. AB - OBJECT: Cervical radiofrequency neurotomy (CRFN) is used in the treatment of patients with chronic pain and disability due to whiplash injury. Confirmation of its efficiency has, however, been based solely on pain and psychological distress factors. The aim of the present study was to extend the assessment of CRFN efficacy by adding other outcome measures to shed light on neuromotor-functional psychological interactions by undertaking comparison of pre- and 1-year postintervention data. METHODS: Forty patients with chronic whiplash injury associated disorders were evaluated prior to and at two separate sessions after CRFN. The evaluation included Neck Disability Index, cervical range of motion, isometric cervical muscle strength, cervical pressure pain threshold, Symptom Check List-90-Revised, and subjective Self-Report of Improvement (SRI). The authors found that the CRFN had a significantly positive effect on all measured parameters. A case-by-case analysis revealed improvement in 70% of the patients at the final follow-up examination. Using stringent cutoff values, between 30 and 60% of the patients experienced measurable improvement. Evaluation of SRI results indicated that more than 80% of the patients were satisfied with the procedure. CONCLUSIONS: Approximately 1 year after intervention, CRFN was associated with an acceptable rate of success, as reflected by objective and subjective outcome measures. PMID- 16703904 TI - Percutaneous drainage and continuous irrigation in patients with severe pyogenic spondylitis, abscess formation, and marked bone destruction. AB - OBJECT: The use of percutaneous suction aspiration has recently become viewed as an effective management strategy for pyogenic spondylitis unresponsive to conservative treatment. What remains unclear is whether it can be effective for severe pyogenic spondylitis in which abscess formation or marked bone destruction is present. The authors undertook a study to clarify answers to this question. METHODS: The authors evaluated clinical and radiographic/neuroimaging data obtained in five patients with severe pyogenic spondylitis, extensive abscesses, and marked bone destruction. These patients had undergone percutaneous drainage and continuous irrigation because open surgery was considered contraindicated in light of their poor general health. The mean period during which continuous irrigation was applied was 9 days (range 7-11 days), and the mean period during which the drainage tube was in place was 19 days (range 13-38 days). All patients suffered from back pain, which was relieved by the percutaneous technique in four patients after a mean of 8 days. The abscesses and inflammation resolved in all patients. Progressive osseous destruction was not observed, and open surgery was performed in only one patient in whom back pain persisted as a result of spinal instability. CONCLUSIONS: After an unsuccessful course of conservative treatment, severe pyogenic spondylitis with abscess formation or marked bone destruction was successfully treated using percutaneous drainage and continuous irrigation. Based on their results, the authors believe that this procedure can be used in patients with severe pyogenic spondylitis that was unresponsive to conservative treatment, particularly in those whose general health is poor. PMID- 16703905 TI - Head-turned rear impact causing dynamic cervical intervertebral foramen narrowing: implications for ganglion and nerve root injury. AB - OBJECT: A rotated head posture at the time of vehicular rear impact has been correlated with a higher incidence and greater severity of chronic radicular symptoms than accidents occurring with the occupant facing forward. No studies have been conducted to quantify the dynamic changes in foramen dimensions during head-turned rear-impact collisions. The objectives of this study were to quantify the changes in foraminal width, height, and area during head-turned rear-impact collisions and to determine if dynamic narrowing causes potential cervical nerve root or ganglion impingement. METHODS: The authors subjected a whole cervical spine model with muscle force replication and a surrogate head to simulated head turned rear impacts of 3.5, 5, 6.5, and 8 G following a noninjurious 2-G baseline acceleration. Continuous dynamic foraminal width, height, and area narrowing were recorded, and peaks were determined during each impact; these data were then statistically compared with those obtained at baseline. The authors observed significant increases (p < 0.05) in mean peak foraminal width narrowing values greater than baseline values, of up to 1.8 mm in the left C5-6 foramen at 8 G. At the right C2-3 foramen, the mean peak dynamic foraminal height was significantly narrower than baseline when subjected to rear-impacts of 5 and 6.5 G, but no significant increases in foraminal area were observed. Analysis of the results indicated that the greatest potential for cervical ganglion compression injury existed at C5-6 and C6-7. Greater potential for ganglion compression injury existed at C3-4 and C4-5 during head-turned rear impact than during head-forward rear impact. CONCLUSIONS: Extrapolation of present results indicated potential ganglion compression in patients with a non-stenotic foramen at C5-6 and C6-7; in patients with a stenotic foramen the injury risk greatly increases and spreads to include the C3-4 through C6-7 as well as C4-5 through C6-7 nerve roots. PMID- 16703906 TI - A novel human osteoblast-derived severe combined immunodeficiency mouse model of bone metastasis. AB - OBJECT: One of the major difficulties of conducting bone metastasis research is the lack of adequate models for studying the bone-tumor microenvironment. The limitations of current in vivo models include the following: non-human tumor or bone, variable reproducibility, limited supply, and an inability to be easily manipulated. The objective of the present study was to develop a uniform and reproducible model of bone/spine metastasis by utilizing bone derived from human osteoblasts grown subcutaneously in severe combined immunodeficiency (SCID) mice with subsequent introduction of human carcinoma cell lines. METHODS: Human osteoblasts were serially passed in culture and induced to differentiate into mature osteoblasts. They were subsequently loaded on hydroxyapatite-coated collagen sponges and implanted subcutaneously into the SCID mice. After allowing the bone to mature for 8 weeks, tumor cell suspensions were implanted percutaneously into the bone. The bone-tumor complexes were subsequently harvested, decalcified, and prepared for histological examination. CONCLUSIONS: The authors have developed a novel, reproducible SCID mouse model of bone/spine metastasis by using bone derived from human osteoblasts and subsequently introduced human tumor lines. They believe this model will be useful for studying the basic biology of bone metastases. PMID- 16703907 TI - Investigation of the dose-dependent neuroprotective effects of agmatine in experimental spinal cord injury: a prospective randomized and placebo-control trial. AB - OBJECT: No definitive treatment for spinal cord injuries (SCIs) exists, and more research is required. The use of agmatine [4-(aminobutyl)-guanidine-NH2-CH2-CH2 CH2-CH2-NH-C(-NH2)(=NH)], a guanidinium compound formed by decarboxylation of L arginine by arginine decarboxylase, is a neurotransmitter-neuromodulator with both N-methyl-D-aspartate receptor (NMDAR)-antagonizing and nitric oxide synthase (NOS)-inhibiting activities. The purpose of this study was to demonstrate the dose-dependent activity of agmatine, an inducible NOS (iNOS) inhibitor and selective NMDAR antagonist, on biochemical and functional recovery in an experimental rat SCI model. METHODS: This study involved 40 Wistar albino male rats. The rats were subjected to sleep-awake cycles for 7 days before surgery. In each group, general anesthesia was induced by a 60-mg/kg ketamine injection. For the surgical SCI model, a Yasargil aneurysm clip was placed in the spinal cord. The study was conducted in the following four main groups: Group I (control group) laminectomy only; Group II, trauma-only group and SCI; Group III, laminectomy, SCI and agmatine 50 mg/kg for 10 days; and Group IV, laminectomy, SCI, and agmatine 100 mg/kg for 10 days. On Day 1, no statistical difference was observed in any group (p < 0.005, analysis of variance [ANOVA] and the Fisher protected least significant difference [PLSD]). On Day 2, no statistical difference was noted among Groups II, III, and IV (p = 0.27, p = 0.42, and p = 0.76, respectively; ANOVA and Fisher PLSD). Beginning on Day 3, recovery in Groups III and IV differed significantly from that in Group II (p < 0.005, ANOVA and Fisher PLSD), and a statistically significant difference between Groups III and IV was observed, which also was present on Days 5, 7, and 10 (p = 0.003, p = 0.0024, and p = 0.0036, respectively; ANOVA and Fisher PLSD). Several observations were noteworthy: motor function scores were reduced significantly in the spinal cord-injured rats compared with the controls (p < 0.005); on Day 1, the agreement of motor function scores in rats in each SCI group indicated that the traumatic event had been replicated equally across all groups (p = 0.59, p = 0.59, and p = 0.28); a statistically significant difference in motor function scores developed on Day 3 between the rats subjected to trauma alone (Group II) and those treated with agmatine (Groups III and IV) (p < 0.005); and no statistically significant intergroup difference in motor function existed at any postinjury interval between the 50- and 100-mg/kg/day agmatine-treated rats (p > 0.005). CONCLUSIONS: Agmatine administration following SCI was shown to reduce NO levels significantly. No statistically significant intergroup difference in the reduction of NO levels was found between rats treated with 50- and 100-mg/kg/day doses of agmatine. Administration of a 100-mg/kg/day dose of agmatine reduced the NO levels to those measured in controls. The authors conclude that with additional studies into the role of agmatine, this drug may be helpful in the treatment of patients with SCIs. PMID- 16703908 TI - Nonmissile penetrating spinal injury. Case report and review of the literature. AB - Nonmissile penetrating spinal injuries (NMPSIs) are rare, even among the population of patients treated in large trauma centers. Patients who present with retained foreign body fragments due to stabbings represent an even smaller subset of NMPSI, and their optimal management is unclear. The authors report the case of a 42-year-old man who presented to the University of California at Davis Medical Center with a retained knife blade after suffering a stab wound to the lower thoracic spine. They discuss this case in the context of a literature review and propose management options for patients with NMPSIs in whom fragments are retained. A search of PubMed was undertaken for articles published between 1950 and 2006; the authors found 21 case reports and eight case series in the English language literature but discovered no published guidelines on the management of cases of NMPSI with retained fragments. After clinicians undertake appropriate initial trauma evaluation and resuscitation, they should obtain plain x-ray films and computerized tomography scans to delineate the anatomical details of the retained foreign body in relation to the stab wound. Neurosurgical consultation should be undertaken in all patients with an NMPSI, whether or not foreign body fragments are present. Surgical removal of a retained foreign body is generally recommended in these patients because the fragments may lead to a worse neurological outcome. Perioperative antibiotic therapy may be beneficial, but the result depends on the nature of the penetrating agent. There is no documentation in the literature to support the use of steroid agents in patients with NMPSIs. PMID- 16703909 TI - En bloc marginal excision of a multilevel cervical chordoma. Case report. AB - The purpose of this case report is to demonstrate that an en bloc resection with negative surgical margins can be successfully achieved in a case of a seemingly unresectable C-2 chordoma if appropriate preoperative staging and planning are performed. The management of chordomas is controversial and challenging because of their location and often large size at presentation. Because chordomas are malignant and will aggressively recur locally if intralesional resection is conducted, wide or true en bloc resection is generally recommended. The literature indicates, however, that surgeons are reluctant to perform wide or even marginal resections because of the lesion's complex surrounding anatomy and the risk of significant neurological compromise when a tumor abuts the dura mater or neural tissues. In this report the authors outline the successful en bloc resection of a large C1-3 chordoma and discuss the importance of preoperative staging and planning. PMID- 16703910 TI - Spondylolisthesis of the thoracic spine. Case report. AB - The thoracic spine is stabilized in the anteroposterior direction by the rib cage and the facet joints. Spondylolisthesis of the thoracic spine is less common than that of the lumbar spine. The authors describe a rare case of thoracic spondylolisthesis in which the patient suffered back pain and myelopathy. The patient was a 44-year-old woman. Plain radiography revealed Grade I T11-12 spondylolisthesis. The pedicle-facet joint angle at T-11 was 118 degrees, greater than that of T-10 or T-12. Postmyelography computerized tomography scanning revealed posterior compression of the dural sac as well as enlargement of and degenerative changes in the facet joint at T-11. Magnetic resonance imaging showed anterior and posterior compression of the spinal cord at the level of the spondylolisthesis. To achieve posterior T10-12 decompression, the surgeons performed a laminectomy and posterolateral fusion in which a pedicle screw fixation system was placed. The patient's back pain disappeared immediately after the operation. The authors conclude that the enlargement of the pedicle-facet joint angle and the degenerative changes of the facet joint caused the thoracolumbar spondylolisthesis. PMID- 16703911 TI - Cervical osteophytes: a cause of potentially life-threatening laryngeal spasms. Case report. AB - This 80-year-old woman presented with acute breathing difficulty during neck flexion when cyanosis also developed. The only potential causes were detected on cervical magnetic resonance imaging: two large anterior cervical osteophytes compressing the retropharyngeal space. Excision of these osteophytes resulted in resolution of the symptoms. PMID- 16703912 TI - Lymphatic fluid fistula: an extremely rare complication of posterior lumbar transpedicular screw fixation. Case report. AB - The authors describe the unique case of a patient who had undergone posterior stabilization of the lumbar spine complicating the course of a lymphatic fistula. A lymphatic fistula is a rare complication of posterior lumbar surgery. Predisposing factors include individual anatomy, scarring adherences due to previous abdominal operations or surgical maneuvers deep in the plane of the transverse processes. Because the onset of lymphatic fistulas is subtle, and because they are associated with a high mortality rate and require multidisciplinary treatment, care is needed to avoid misdiagnosing these lesions as the more common cerebrospinal fluid fistula. PMID- 16703913 TI - Primary clear-cell chondrosarcoma of the cervical spine. Case illustration. PMID- 16703914 TI - Isolated Rosai-Dorfman disease of the sacrum. Case illustration. PMID- 16703916 TI - Magnet designation: the mark of nursing excellence. PMID- 16703915 TI - Pathogenesis of tumor-associated syringomyelia demonstrated by peritumoral contrast material leakage. Case illustration. PMID- 16703917 TI - Medical robotics: the impact on perioperative nursing practice. AB - Robotic technology and the increased use of minimally invasive surgery approaches is altering the environment in which operating room personnel work and affecting how nurses must care for patients. An understanding of the history of robotics, current applications of the technology, and perioperative nursing responsibilities is needed to assure quality patient care in the wake of continued advances in technology. PMID- 16703918 TI - Care of the patient undergoing robotic-assisted laparoscopic pyeloplasty. AB - Laparoscopic pyeloplasty as a treatment for ureteropelvic junction obstruction has shown comparable success rates with open pyeloplasty techniques. The use of robotic technology to assist during laparoscopic pyeloplasty procedures has been encouraged by the steep learning curve for laparoscopic surgical skills, and the complexity of laparoscopic suturing. Robotic technology provides the surgeon with the ability to filter out any physiologic hand tremor, more degrees of freedom of movement than traditional laparoscopic instruments, the ability to scale movement to provide better control for microsurgery, better ergonomics during surgery, and three-dimensional vision. Details of the procedure and specific nursing care of the patient undergoing robotic-assisted laparoscopic pyeloplasty at the University of Iowa Hospital and Clinics are described. PMID- 16703919 TI - Care of the patient undergoing radical cystectomy with a robotic approach. AB - Radical cystectomy or cystoprostatectomy with urinary diversion is the gold standard for the treatment of muscle-invasive bladder cancer. Cystectomy can be through an open or robotic-assisted laparoscopic approach. Advances in laparoscopy, robotic surgery, and urological oncology have made it possible for select surgeons to perform nerve-sparing robotic-assisted laparoscopic radical cystoprostatectomy. Advantages of robotic surgery may be minimal blood loss, shorter hospital stay, quicker recovery, and possibly more precise and rapid removal of the bladder depending on the experience and expertise of the surgeon. Appropriate patient selection and thorough pre-operative evaluation, however, are key in maximizing positive surgical outcomes. The experience at the University of Virginia with robotic-assisted laparoscopic radical cystectomy will be discussed. PMID- 16703920 TI - Robotic-assisted laparoscopic cystectomy with ileal conduit for urinary diversion. AB - This case demonstrates the need for flexibility when approaching major abdominal surgery through a robotic approach. While the case was initiated with robotic technology, it ultimately had to be converted to an open procedure for reasons of patient safety and to ensure a good surgical outcome. PMID- 16703921 TI - Care of the patient undergoing robotic-assisted prostatectomy. AB - Prostate cancer has many treatment options. In addition to open retropubic and perineal approaches to radical prostatectomy, laparoscopic robotic prostatectomy is available as a newer surgical option. Potential advantages of robotic surgery include reduced pain and trauma, less blood loss, reduced infection risk, shorter hospital stay, faster recovery, and less scarring (Intuitive Surgical, 2005). A variety of nursing care considerations involving pre-operative education and preparation, intra-operative and immediate postoperative care issues, and long term followup must be understood to meet the needs of a robotic surgical patient. Patient selection is very important to optimize a positive surgical outcome. Just as certain criteria make a good surgical candidate, there are factors that could complicate the surgery or adversely affect recovery. PMID- 16703922 TI - Robotic prostatectomy surgery. AB - Despite the minimally invasive nature of the robotic prostatectomy procedure, there are still many cultural, environmental, and social issues that must be addressed. The following case study emphasizes that patient education is key to successful outcomes following robotic prostatectomy. PMID- 16703924 TI - Getting ready for certification: pediatric urology, advanced practice, and acute scrotal pain. PMID- 16703923 TI - Update on preparing for the next influenza pandemic. AB - While previous pandemics have taken the world by surprise, the present situation has given the world advanced warning and a rare opportunity to prepare for the next pandemic. WHO urges all countries to take action now to assess their level of preparedness, identify priority needs, and develop a national influenza preparedness plan. If containing or delaying the spread of a new human influenza strain is possible, it will take rapid, massive coordinated global and national actions. The success of these actions will depend on the preparations that have made in advance by each member of the international community. PMID- 16703925 TI - Cholesterol, cholesterol lowering agents/statins, and urologic disease: part I - knowing your numbers. AB - Educating patients on the basic cardiovascular disease (CVD) risk markers such as cholesterol can be difficult in any medical setting, but especially in urology where patients are being evaluated for non-cardiovascular conditions. Primary reasons for discussing cholesterol or cardiac risk factors and assessment in urology include (a) the primary cause of death of men and women in the United States and most countries around the world is CVD; (b) the primary or secondary cause of death from the largest cancer prevention trials (high or average risk) is CVD; (c) the primary or secondary cause of death in men with prostate cancer is CVD; (d) there may be a correlation in some of the mechanisms that contribute to CVD and mechanisms that contribute to numerous urologic conditions, such as benign prostatic hyperplasia, bladder cancer, erectile dysfunction, female sexual dysfunction, kidney cancer, and prostate cancer; and (e) one of the better methods to monitor the success of lifestyle changes for the patient in urology is to monitor these CVD markers, as is the case in some lifestyle studies of men with prostate cancer. PMID- 16703926 TI - Where will the next generation of special needs dentists come from? PMID- 16703927 TI - Effect of reinforced laryngeal mask airway or endotracheal intubation anesthesia on adverse events: Quality of life issues for dental patients requiring general sedation. PMID- 16703928 TI - Bisphosphonate induced osteochemonecrosis of the jaws: an ounce of prevention may be worth a pound of cure. AB - Patient exposure to bisphosphonate drugs for the management of hypercalcemia of malignancy, osteolytic lesions of metastatic cancer and osteoporosis has led to increasing reports of osteochemonecrosis of the jaws (bis-phossy jaw). This serious and debilitating condition requires dental practitioners to be alert for signs and symptoms of this syndrome. Thus far, nitrogen containing bisphosphonates have been implicated as a causative agent. While only a small fraction of patients who have taken these agents will develop osteochemonecrosis, it seems that patients who have received intravenous bisphosphonates are at greater risk than those who have taken oral agents. Tooth extractions are the most frequently reported predisposing dental procedure. While appropriate management strategies for patients with osteochemonecrosis of the jaws are evolving, we are suggesting rational preventive protocols and therapies based upon current experience and knowledge. These recommendations may change over time as the profession gains more experience in managing these patients. PMID- 16703929 TI - A comparison of the gingival health of children with Down syndrome to healthy children residing in an institution. AB - The purpose of this study was to compare the onset and severity of gingivitis in children with Down syndrome, when compared to a healthy control group of children. The subjects included 41 children with Down syndrome ages two to 14 years (mean age: 7.6 years) and 112 age-matched healthy controls. We assessed the gingival health of all subjects using the gingival inflammation (M-PMA) index and periodontal probing depth (PD). Children were divided into three age categories: <5 years (AI), 5 to <10 years (AII), and 10 to <17 years (AIII). Supragingival plaque was measured using the Oral Hygiene Index (OHI) and the subjects were screened with the BANA test (Perioscan-Oral-B). Measurement of the M-PMA index in the healthy children showed an age-related increase (F = 10.369, p < 0.001), and the M-PMA index at the younger age group <5 year (AI) was significantly lower than that for the other two age groups All or AIII (p < 0.005, p < 0.001). In contrast, the M-PMA index values at AI and AIII in the subjects with Down syndrome were significantly higher than those for healthy children (p < 0.001, p < 0.001). Both groups had an age-related increase in PD (F = 3.388, p < 0.05 & F = 10.806, p < 0.001), and PD at AIII was significantly higher than that at AI in both groups (p < 0.01, p < 0.001). The children with Down syndrome showed an age related increase in the BANA test score (F = 3.452, p < 0.05), and the BANA test score at AIII was significantly higher than that at AI (p < 0.02). The BANA test score in the healthy children was not age-related but was significantly higher than that in the children with Down syndrome (p < 0.02, p < 0.05). PMID- 16703930 TI - Dental care management of a young patient with extensive lymphangioma of the tongue: a case report. AB - Lymphangiomas are benign tumors resulting from a congenital malformation of the lymphatic system. Relatively uncommon, lymphangiomas are usually diagnosed at birth and develop within the first years of life. When these tumors occur in the oral cavity, the tongue is the most frequently affected site. Lymphangioma of the tongue is a common cause of macroglossia in children, which may lead to a dry/cracked tongue with ulcerating secondary infections, difficulty in swallowing and mastication, speech disturbances, exclusive nasal breathing, airway obstruction, mandibular prognathism and other possible deformities of maxillofacial structures. This paper discusses the most relevant features, clinical manifestations, disease-related impairments and treatment options for lymphangioma of the tongue. It presents the case report of a five-year-old child diagnosed with this lesion, including a description of the patient's dental care management. PMID- 16703931 TI - A new classification system for dental treatment under general anesthesia. AB - The provision of comprehensive care for patients with special needs using dental general anesthesia (DGA) has changed over time, and now includes more complex procedures and the participation of many services. As a result, it is necessary to integrate, organize and describe all of the procedures that are carried out in different DGA settings. The aim of this study was to propose a systematic classification for dental treatment procedures be delivered under DGA, and to compare this classification system with an existing system. This new classification system has three distinct components: type, frequency and length of time needed to complete dental procedures for both primary and permanent teeth. A wide range of oral surgery procedures and endodontic treatment was also included. A retrospective cohort study utilizing 84 subjects was used to develop and compare the two classification systems. When comparing the different categories of procedures by both classifications, there were significant statistical differences between them (p < 0.05). Oral health care for patients with special needs has evolved, with more complex and extensive interventions that require teamwork by personnel from different dental or medical specialties. The classification system in this study includes detailed information regarding the procedures involved in the DGA. This helps to provide a clear understanding and specific information that enables the comparison of clinical experiences across populations where a DGA has been used for patients with special needs. PMID- 16703932 TI - Dentists' attitudes and practices toward provision of orthodontic treatment for children with visual and hearing impairments. AB - The objective of this study was to determine differences in behavior and attitudes of dentists in Riyadh, Saudi Arabia, in providing orthodontic care for children who are sensory impaired. A self-administered questionnaire was sent to all dentists working in Riyadh to assess the following domains: personal characteristics of the dentists and their practices, provision of dental care for children who are visually-impaired (Vl) and/or hearing-impaired (HI), and their attitude toward providing orthodontic care for these children. Attitudes were measured on two scales and the overall score of these two scales represented each respondent's attitude. Thirty percent of the dentists provided dental care for children with VI and 45.3 percent did for children with HI. The provision of orthodontic care was significantly affected by the country in which the dentists had received their dental training, both for children with VI and HI (p < 0.01), and by number of years they had been in practice for children with VI (p < 0.05). Regression analysis showed that only the country of dental training significantly affected the dentist's attitude score. There were also significant variations in attitudes toward the provision of orthodontic treatment for children with sensory impairment (SI), influenced by dental training and experience. In practical terms, this means that improvement in attitudes needs to be initiated at the dental undergraduate level. Establishing global guidelines for the provision of orthodontic treatment for patients with sensory impairment is likely to assist both professionals and patients. PMID- 16703933 TI - The use of botulinum toxin-a in the treatment of severe bruxism in a patient with autism: a case report. AB - This case report describes an alternative method for reducing bruxism in a special needs patient who was not a candidate for an intraoral appliance. Bruxism is often seen in patients with special needs and can result in excessive dental wear, temporo-mandibular joint pain, avulsion of teeth and other problems. Current methods of management are not typically effective in this population because most require patient compliance. An 11-year-old male diagnosed with autism and Bannayan-Zonana syndrome received bilateral injections of botulinum toxin type-A (Botox Allergan Pharmaceuticals, Irvine CA) in the masseter muscle. The patient's condition was followed up via post-operative telephone interviews with the parents for 60 days. A reduction in the frequency and severity of bruxism was reported. The only side effects noted were soreness at the injection site and mild, temporary drooling. Although further research is required to determine the optimal doses and injection frequency, botulinum toxin type-A appears to be an alternative method for controlling bruxism in the special needs population. PMID- 16703934 TI - Dental implants in edentulous adults with cognitive disabilities: report of a pilot project. AB - Individuals with cognitive disabilities face many barriers to oral care, often suffering from partial or complete edentulism. While the use of implant reconstruction is becoming more common in the general population, such care is still being used infrequently in individuals with intellectual impairment. A pilot project in 1995 surgically placed and restored implant-supported prostheses in six edentulous adults who had varying degrees of cognitive impairment. This report presents the dentists' reconstruction experiences and the patients' follow up care, and discusses the results in relationship to current literature. Experiences from these patients suggest that behavior during the restorative process, prosthetic complications post placement, and patients' oral hygiene practices should influence patient selection and prosthetic design. Anteriorly placed fixtures and removable designs, which make self-care and repair easier, can be used to treat an edentulous population. PMID- 16703935 TI - Marginal zone B-cell lymphoma of the uterus: a case report and review of the literature. AB - We report here a case of a 52-year-old female in whom immunohistological studies of the uterus established a diagnosis of extranodal marginal zone B-cell lymphoma. Malignant lymphoma arising from mucosa-associated lymphoid tissue (MALT) of uterus is extremely rare. Accurate histologic interpretation of uterine lymphoma is essential, as treatment options and prognosis vary based on the histological grade of such lymphomas. Patients with primary uterine lymphoma generally have intermediate or high-grade lymphoma and poorer prognosis. PMID- 16703936 TI - Lung abscess caused by Clostridium ramosum. AB - A 49-year-old male presented with weight loss and fever. Further evaluation revealed lung abscesses and Clostridium ramosum bacteremia. This is an unusual cause of infection. There have only been seven reports of infections caused by C. ramosum. PMID- 16703937 TI - Managing risk by avoiding peer review pitfalls. PMID- 16703938 TI - An unusual case of GI bleeding diagnosed by capsule endoscopy. PMID- 16703939 TI - Anatomy of the avian hippocampal formation. AB - Increasing knowledge of the avian hippocampal formation (hippocampus and parahippocampal area) suggests that it plays a role in a variety of behaviors, such as homing, cache retrieving, visual discrimination, imprinting, and sexual behavior. Knowledge of the neural circuits in the hippocampal formation and its related areas or nuclei is important for the understanding of these functions. This review therefore describes the functional neuroanatomy of the avian hippocampal formations, i.e., its subdivisions, cytoarchitecture, and afferent and efferent connections. Evidence obtained by a combination of Nissl staining and tract-tracing shows that the pigeon hippocampal formation can be divided into seven subdivisions: dorsolateral (DL), dorsomedial (DM), triangular (Tr), V shaped (V), magnocellular (Ma), parvocellular, and cell-poor regions. DL and DM can be further divided into dorsal and ventral, and lateral and medial portions, respectively. In the hippocampal formation, reciprocal connections are found between DL-DM, DL-Tr, DL-Ma, DM-Ma, DM-V, and Tr-V. Neurons in the V-shaped layer appear to be intrinsic neurons. Sensory inputs from higher order visual and olfactory stations enter DL and DM, are modified or integrated by intrinsic hippocampal circuitry, and the outputs are sent, via DL and DM, to various telencephalic nuclei, septum, and hypothalamus. The neural pathways indicate that the hippocampal formation plays a central role in the limbic system, which also includes the dorsolateral corticoid area, nucleus taeniae of the amygdala, posterior pallial amygdala, septum, medial part of the anterior dorsolateral nucleus of the thalamus, and the lateral mammillary nucleus. Connectional and comparative studies, including the use of kainic acid excitotoxicity, suggest that the V-shaped layer is comparable to the dentate gyrus of the mammalian hippocampal formation and DM to Ammon's horn and subiculum. PMID- 16703941 TI - Effects of partial hippocampal lesions by ibotenic acid on repeated acquisition of spatial discrimination in pigeons. AB - Pigeons were trained on a spatial discrimination task using a repeated acquisition procedure. In this procedure, the pigeons were trained to discriminate between the positions of three keys. One of them was designated the correct key. When the subjects reached the criterion, the discrimination task was changed, with one of two previously incorrect keys now being made the correct key. This procedure was repeated at least 15 times. Then, lesions to the whole hippocampus, the medial hippocampus or to the lateral hippocampus were made by injections of ibotenic acid (Experiment 1). Only the subjects with damage to the whole hippocampus showed deficits in learning after the lesions. The deficits were similar to those caused by aspiration lesions /37/. Knife cuts separating the medial and lateral hippocampi were made in Experiment 2. The subjects did not show deficits in the spatial discrimination task after the sections. Although studies of the connectivity in the avian hippocampus suggested functional differences between the medial and lateral hippocampi, the present results show that pigeons can learn spatial discrimination with the medial and lateral parts of hippocampus separated. PMID- 16703940 TI - Representing the richness of avian spatial cognition: properties of a lateralized homing pigeon hippocampus. AB - Brain organization and its relationship to behavior in any extant species is a reflection of a long evolutionary history of adaptive change. Therefore, it follows that the relationship between the hippocampus and spatial cognition in any species or taxonomic group would be characterized by features adapted to its spatial ecology. Birds are the animal world's supreme navigators, and aspects of their navigational ability are dependent on the integrity of the hippocampal formation. Using the homing pigeon as a model species, we review an accumulating body of data indicating that the avian hippocampus is functionally lateralized. The spatial response properties of left hippocampal neurons, as recorded in freely moving pigeons in a laboratory environment, differ from the response properties of right hippocampal neurons. Left hippocampal lesions generally disrupt navigational behavior under field conditions more than right lesions, while right lesions are more likely to disrupt goal localization behavior under laboratory conditions. We propose that the available data are consistent with a hypothesis of a left hippocampus more involved in navigational processes, and a right hippocampus more involved in representing the locations of events. We also discuss the extent to which the observed hippocampal lateralization should be viewed as an intrinsic property of the hippocampus itself or imposed by the lateralized properties of visual inputs originating in other brain regions. Whatever the nature of the observed hippocampal lateralization, it is likely one adaptive variation in hippocampal organization that supports the extraordinary spatial behavior of birds. PMID- 16703942 TI - Spatial memory and hippocampal function in a non-foodstoring songbird, the zebra finch (Taeniopygia guttata). AB - Spatial memory and hippocampal function have as yet been investigated mainly in pigeons and food storing songbirds. We show here that the zebra finch, a songbird not specialized in food storing and caching, is also able to learn a spatial memory task and uses a spatial map for finding food in a 'dry water maze'. Hippocampal lesions prevent learning and retention of this spatial task. The immediate early gene (IEG) products Zenk and Fos are expressed within the hippocampus when the bird is learning the task. Spatial learning cannot be assigned to any hippocampal subregion; IEG expression within the hippocampus is patchy and seems almost arbitrarily located. The IEG activation pattern in spatial memory experiments is compared with those in other learning experiments with zebra finches. PMID- 16703943 TI - A multi-disciplinary approach to understanding hippocampal function in food hoarding birds. AB - Spatial memory and the hippocampal formation (HF) of food-hoarding birds have been put forward as a prime example of how natural selection has shaped a cognitive system and its neural underpinnings. Here, I review what we know about the HF of hoarding birds and lay out the work that is currently underway to use this system to obtain a better understanding of hippocampal function in general. This interdisciplinary programme includes evolutionary, ecological, psychological, ethological, and neuroscientific approaches to the study of behaviour and cognition. Firstly, we need to understand the behaviour of the birds in their natural environment, and identify the aspects of cognition and behaviour that may be especially valuable for the species under study. Secondly, these cognitive and behavioural traits are compared to closely-related non hoarding species. Thirdly, we also compare HF anatomy between closely-related hoarding and non-hoarding species, identifying possible neural mechanisms underlying behavioural differences. Finally, behavioural and neuroscientific approaches are combined in experiments directly investigating the involvement of the HF or any of its anatomical and physiological aspects in the behaviours under study. This process loops back upon itself in many different ways, with all the different approaches informing each other. In this way we are making progress in understanding the functioning of the HF, not only in food-hoarding birds, but in all vertebrates. PMID- 16703945 TI - From movement to transitivity: the role of hippocampal parallel maps in configural learning. AB - Whether spatial learning is a special case of configural or relational learning, or whether abstract principles evolved from the concrete need to navigate in space, is a question of long-standing debate. The parallel map theory of hippocampal function offers a resolution of the debate by redefining 'spatial learning' as two parallel, geometric processes, Euclidean metric and topological. Moreover, these processes are subserved by independent hippocampal subfields that underlie two ways of representing space, the bearing and the sketch map. It is possible that configural and relational learning, like spatial learning, should also be distinguished in this way. Transitive inference, requiring the construction of a value gradient, could be analyzed as a Euclidean metric problem. In contrast, transverse patterning could be seen as a topological analysis of the relationships among discrete objects. If this interpretation is correct, lesions to the primary bearing map structure (dentate gyrus) should impair transitivity while lesions to the primary sketch map structure (CA1) should impair transverse patterning and similar topological tasks. Recent results from diverse species and tasks lend support to these predictions, suggesting that the hippocampus not only creates parallel maps but uses these maps to solve more abstract configural or relational problems. PMID- 16703946 TI - Rearing on hind legs, environmental novelty, and the hippocampal formation. AB - Many mammals spontaneously rear on their hind legs in response to novelty. The current paper is the first review of rearing behaviour, and is intended to collate findings from different perspectives that are not usually brought together. We suggest that rearing is a useful marker of environmental novelty, that the hippocampal formation is a crucial component of the system controlling rearing in novel environments, and that rearing is one of several ethological measures that can profitably be used to assess hippocampal learning and memory. Consideration is given to the following topics: the possible functions of rearing in information-gathering and escape behaviour; the modulation of rearing by various factors such as anxiety/ fear emotionality; comparative perspectives on rearing; neuroanatomical circuits involved in rearing with particular reference to the hippocampal formation and its afferents and efferents; and the role of the hippocampal formation in uncharted and mismatch environmental novelty. The review concludes with testable predictions about rearing, environmental novelty and the hippocampus. PMID- 16703944 TI - The boundary vector cell model of place cell firing and spatial memory. AB - We review evidence for the boundary vector cell model of the environmental determinants of the firing of hippocampal place cells. Preliminary experimental results are presented concerning the effects of addition or removal of environmental boundaries on place cell firing and evidence that boundary vector cells may exist in the subiculum. We review and update computational simulations predicting the location of human search within a virtual environment of variable geometry, assuming that boundary vector cells provide one of the input representations of location used in mammalian spatial memory. Finally, we extend the model to include experience-dependent modification of connection strengths through a BCM-like learning rule - the size and sign of strength change is influenced by historic activity of the postsynaptic cell. Simulations are compared to experimental data on the firing of place cells under geometrical manipulations to their environment. The relationship between neurophysiological results in rats and spatial behaviour in humans is discussed. PMID- 16703947 TI - Effects of post-training lesions in the hippocampus and the parietal cortex on idiothetic information processing in the rat. AB - Dead reckoning can be defined as the ability to navigate using idiothetic information based on self-movement cues without using allothetic information such as environmental cues. In the present study, we investigated the effects of hippocampal and parietal cortex lesions on homing behavior using dead reckoning in rats. Experimentally naive Wistar rats were trained with a homing task in which rats were required to take a food pellet from a cup in the arena and to return home with the pellet. After training, rats were divided into a control (CONT) group (n = 16), hippocampal lesioned (HIPP) group (n = 16), and parietal cortex lesioned (PARC) group (n = 16), and rats in the lesioned groups underwent surgery. After surgery, Test 1 (with four cups) and Test 2 (with one cup but the outgoing path was diverted by a barrier) were conducted. The HIPP group showed severe impairment in homing, but the performance of the PARC group did not differ from that of the CONT group. HIPP rats either approached wrong doors or ate the pellet in the arena. Circular statistics showed that homing directions of CONT and PARC rats showed concentration towards home, whereas those of HIPP rats did not. Our results exhibiting HIPP rats' failure in homing agree with many previous studies, but the results obtained from PARC rats were different from previous studies. These results indicate that the intact hippocampus is important for dead reckoning, but the role of the parietal cortex in dead reckoning is still not clear. PMID- 16703948 TI - Effects of hippocampal lesions on conditional discriminations in rats. AB - Effects of hippocampal lesions on conditional discrimination were examined in rats. Rats were trained in a conditional drug discrimination paradigm using pentobarbital. They were able to learn the discrimination. After hippocampal lesions, all rats maintained discrimination. Thus a hippocampal lesion had no effect on retention of the conditional discrimination with drugs. Another group was trained in the same conditional discrimination except that the conditional stimuli were two different flashing lights. Similarly to the drug discrimination, only one of the two light conditioning stimuli was presented in one training session. No rat was able to learn the discrimination. New rats were trained in conventional conditional discrimination, in which the two conditional stimuli (flashing lights) were repeatedly presented within one training session. Hippocampal lesions made after learning this discrimination had no effect on retention. The present study reveals that the hippocampus does not have a crucial role in conditional discrimination learning when conditional stimuli were drugs and that rats could not learn conditional discrimination when two external conditional stimuli were presented in a similar way to the internal stimuli (drug discrimination paradigm). PMID- 16703949 TI - Timing and hippocampal theta in animals. AB - All animals have at least two different internal clocks, one governing cognition of time of day, and the other concerning awareness of seconds and minutes. In the latter case, organisms show scalar properties. The timing mechanisms in the brain may function similarly throughout the animal kingdom, but this is not yet clear. Previous studies have shown that the hippocampus is intricately involved with the process of interval timing. Data concerning electrophysiological field potentials in the hippocampus show obviously rhythmic activity, known as hippocampal theta activity. An information-processing model of interval timing postulates three distinct stages: a clock, a memory, and a decision stage /11/. The timing process includes memory processing, which means that the hippocampus works together with working memory to estimate current time passing. PMID- 16703950 TI - Dissociation of the roles of NMDA receptor and hippocampus in rats' spatial learning: the effects of environmental familiarity and task familiarity. AB - In this study, we compared the effects of MK-801 and hippocampal lesions on re training of Morris water maze place task in familiar and novel environments in rats. In Experiment 1, rats were pre-trained with the place task. After acquiring the task, rats were re-trained with the same task in a familiar environment following MK-801 injection, and were then trained with the same task in a novel environment following MK-801 injection. In the familiar environment, MK-801 had no effect, but in the novel environment performance was impaired. In Experiment 2, after the place task training, the hippocampus was lesioned, and rats were re trained with the same task in the familiar environment then retrained again in the novel environment. Rats showed severe impairment in both environments. These two experiments suggest different functions for NMDA receptors and the hippocampus. The results of Experiment 1 showed that NMDA receptors are not required for utilizing spatial representations but they play an important role in the construction of spatial representations. The results of Experiment 2 show that the hippocampus is necessary for both the utilization of spatial representations already formed and the formation of new spatial representations. PMID- 16703951 TI - Spatial view cells in the primate hippocampus and memory recall. AB - Hippocampal spatial view neurons in primates provide allocentric representations of a view of space 'out there'. The responses depend on where the monkey is looking; and can be updated by idiothetic (self-motion) inputs provided by eye movements when the view is hidden. In a room-based object-place memory task, some hippocampal neurons respond to the objects shown, some to the places viewed, and some to combinations of the places viewed and the objects present in those locations. In an object-place recall task when the location in space at which an object has been seen is recalled by the presentation of the object, some primate hippocampal neurons maintain their responding to the object recall cue in a delay period without the object visible while the place is being recalled; and other neurons respond to the place being recalled. Other spatial view neurons form associations with the rewards present at particular locations in space. These findings, and computational models of the hippocampus, help to show how the primate including human hippocampus is involved in episodic memory. PMID- 16703952 TI - Impairments in recognition memory for object and for location after transient brain ischemia in monkeys. AB - Using an object recognition or location memory test employing either small or large sets of training stimuli, we examined the effects of selective damage to the hippocampus after ischemia in Japanese monkeys. Ischemic (ISC) monkeys were significantly impaired in learning a delayed matching-to-sample (DMS) task (10 sec) when the sample and test objects were drawn from a set of 45 possible stimuli, but not in learning a DMS when a set of 300 possible stimuli was used ISC monkeys were also impaired in a delayed matching-to-location (DML) task that employed a 3-well tray but not in one that employed a 10-well tray. After criterion learning was attained, ISC monkeys were impaired significantly only in the 300-stimuli version and at the longest delay tested (10 min). These results suggest that the hippocampus might be involved in processing comparisons and forming relationships between current and recent stimuli, but not in the remembering of familiar objects, and in long-term maintenance (more than 10 min) of stimulus memory. PMID- 16703953 TI - Transplantation of cryopreserved human bone marrow-derived multipotent adult progenitor cells for neonatal hypoxic-ischemic injury: targeting the hippocampus. AB - There is currently no treatment for neonatal hypoxic-ischemic (HI) injury. Although limited clinical trials of stem cell therapy have been initiated in a number of neurological disorders, the preclinical evidence of a cell-based therapy for neonatal HI injury remains in its infancy. Stem cell therapy, via stimulation of endogenous stem cells or transplantation of exogenous stem cells, has targeted neurogenic sites, such as the hippocampus, for brain protection and repair. The hippocampus has also been shown to secrete growth factors, especially during the postnatal period, suggesting that this brain region presents a highly conducive microenvironment for cell survival. Based on its neurogenic and neurotrophic factor-secreting features, the hippocampus stands as an appealing target for stem cell therapy. In the present study, we investigated the efficacy of intrahippocampal transplantation of multipotent adult progenitor cells (MAPCs), which are pluripotent progenitor cells with the ability to differentiate into a neuronal lineage. Seven-day old Sprague-Dawley rats were initially subjected to unilateral HI injury, that involved permanent ligation of the right common carotid artery and subsequent exposure to hypoxic environment. At day 7 after HI PMID- 16703954 TI - Spatial memory activation of the parietal cortex measured with near-infrared spectroscopic imaging in the finger-maze of the Morris water maze analogue for humans. AB - To assess the cortical regions involved in learning spatial navigation, changes in oxyhemoglobin volume were measured with near-infrared spectroscopy in a task analogous to the Morris water maze. The blindfolded participants (Ps) were allowed to search the area inside the circular ring (20 cm) until they reached a goal (1 cm). The sitting and starting positions were randomly varied across trials, but the goal was in a fixed position so that the Ps could learn the allocentric spatial relationship between the goal and the experimental room. There was a significant sex difference (p < 0.05) in the present task, showing that performance in males was better than that in females. With this task, we measured brain activation in the first training trial, the probe test, and the motor-control trials. In the first trial, dorsolateral prefrontal and parietal cortices were activated. In the probe test, only the dorsolateral prefrontal regions were activated. These results suggest that the parietal cortex is critical for acquisition of spatial memory. PMID- 16703955 TI - Impaired allocentric spatial memory underlying topographical disorientation. AB - The cognitive processes supporting spatial navigation are considered in the context of a patient (CF) with possible very early Alzheimer's disease who presents with topographical disorientation. Her verbal memory and her recognition memory for unknown buildings, landmarks and outdoor scenes was intact, although she showed an impairment in face processing. By contrast, her navigational ability, quantitatively assessed within a small virtual reality (VR) town, was significantly impaired. Interestingly, she showed a selective impairment in a VR object-location memory test whenever her viewpoint was shifted between presentation and test, but not when tested from the same viewpoint. We suggest that a specific impairment in locating objects relative to the environment rather than relative to the perceived viewpoint (i.e. allocentric rather than egocentric spatial memory) underlies her topographical disorientation. We discuss the likely neural bases of this deficit in the light of related studies in humans and animals, focusing on the hippocampus and related areas. The specificity of our test indicates a new way of assessing topographical disorientation, with possible application to the assessment of progressive dementias such as Alzheimer's disease. PMID- 16703956 TI - Memory impairment and awareness of memory deficits in early-stage Alzheimer's disease. AB - Recognition memory of auditory verbal learning tests and awareness of memory deficits were examined in 24 individuals with early-stage Alzheimer's disease (AD) using a performance prediction-postdiction paradigm. Individuals with AD displayed impaired recognition memory, and recognition performance correlated positively with regional cerebral blood flow at rest in bilateral prefrontal areas and the left medial temporal area. In addition, underawareness of memory deficits was also marked even at this early stage. Individuals with AD retrospectively overestimated memory performance after actual performance, but appeared to benefit from feedback, and displayed intact online awareness of memory dysfunction, leading to normal prediction of the second session. However, individuals with AD failed to retrospectively incorporate incidents of memory failure into generalized self-belief systems. Brain/ behavior correlational analyses suggest that the prefrontal cortex and posterior dorsomedial regions including the precuneus may be involved in self-awareness. PMID- 16703958 TI - [Nutritional policy and its framework in Colombia]. AB - OBJECTIVE: Assessing the nutritional policy formulated in the 1996-2005 Colombian Food and Nutrition National Plan (FNNP) using key informants (KI), policy-makers and civil servants. MATERIAL AND METHODS: A descriptive cross-sectional structured survey of 77 KI: 17 policy-makers and 60 civil-servants from PNAN. VARIABLES STUDIED: determinant food factors, the existence of a nutritional policy, assessing policies involved with food security and organisational variables implicit in the policies. A Position Index (PI) was constructed for quantifying KI assessment (0-0.33 = positive evaluation, 0.34-0.67 = partial/readjusted, 1 = 0.68-1= negative evaluation). RESULTS: 79% of KI coincided in stating that there was a Nutritional Policy but that it should be readjusted (IP = 0.50 policy-makers, IP = 0.54 civil servants). KI did not agree about institutional coordination whilst policy-makers said that there was coordination between a reduced group of institutions, including themselves (IP = 0.33); civil servants said that there was no coordination between any of the institutions (IP = 0.75). They also said that the research strategy had been unsuccessful (IP = 0.73). CONCLUSIONS: Ten years after the Nutritional Policy was introduced into Colombia KI say that it should be readjusted. Strategies such as coordination and research could be optimised so that their objectives can be reached. PMID- 16703957 TI - Dissociative contributions of medial temporal and frontal regions to prospective remembering. AB - Prospective memory is memory of future intentions or plans in everyday life. Although some previous neuropsychological studies have stated that the prefrontal cortex and the medial temporal lobe are essential for successful prospective remembering, how the two regions contribute to prospective remembering remains unclear. We therefore used prospective memory training in the present study to investigate the neural mechanism of two components of prospective remembering: remembering to remember and remembering content. Two brain-damaged patients participated in this study: patient Y.O., who had lesions in the medial temporal lobe bilaterally, and patient T.K., who had a lesion in the basal forebrain and right medial frontal lobe. Both participants exhibited a severe anterograde amnesic syndrome and had normal IQ scores. Before the prospective memory training the participants underwent several index memory tests to examine their general memory and prospective memory performance. The training consisted of requesting the participants to perform an original mini-day task, in which they were first asked to memorize five simple daily actions with their times for execution, and then to recall the content of the actions when shown a drawing of a clock showing the proper time for execution. A training session was carried out once a week for 3 months. After completing training, the participants were again requested to take the same index memory tests. The results of the training task showed that Y.O.'s memory performance had gradually improved across all sessions of training, but T.K.'s improvement was not as marked. A more detailed analysis revealed that Y.O.'s memory performance was better for recalling time than for recalling content, whereas T.K.'s memory performance was better for recalling content than for recalling time. Furthermore, the results of the final index memory tests showed that the only improvement in Y.O.'s prospective memory was in remembering to remember, and that the only improvement in T.K.'s prospective memory was in remembering content. These results provide strong evidence that these two remembering components of prospective memory have independent neural bases, with the basal forebrain and right medial frontal lobe being required for remembering to remember, and the medial temporal lobe being required for remembering content. The results also suggest that memory training is an effective, means of improving everyday memory. PMID- 16703959 TI - Introducing a clinical-behavioural scoring system for children's oral hygiene. AB - OBJECTIVES: Developing and testing a clinical-behavioural scoring system for assessing children's oral hygiene. MATERIALS AND METHODS: One clinical variable (the presence of dental plaque, measured using Silness and Loe's index) and one behavioural variable (self-reported tooth brushing frequency) were combined into secondary data analysis of research databases for 3-6-year-olds and 6-13-year olds in a Mexican community. The combined scoring is an ordinal scale that depicts suitable, moderate and inadequate hygiene. Blinded dental examiners also collected dmft/DMFT data in standardised conditions. Data was analysed with Spearman's rho, Kruskall-Wallis, non-parametric tests for trends and Pearson's chi2 tests. RESULTS: 1303 children aged 3-6 years old and 1644 children aged 6-13 years old participated in the study. Clear relationships existed between the combined scoring system and dmft (p < 0.01) and between the scoring system and DMFT (p < 0.01), suggesting that the combined clinical-behavioural scoring system is a reasonably accurate measurement of the relationship between caries experience and oral hygiene in children in the given setting. CONCLUSIONS: The combined clinical-behavioural scoring system is a simple, easy-to-use tool that incorporates clinical and behavioural data commonly found in dental systems. Whether the clinical-behavioural scoring system can be generalised remains to be established. PMID- 16703960 TI - [Scientific production in health science in Colombia, 1993-2003]. AB - OBJECTIVE: Analysing scientific production related to health research between 1993 to 2003 in Colombia to generate information allowing a more focused investment in public health research. METHODS: Bibliometric indicators were used to analyse scientific production registered in MEDLINE and LILACS. Trends in the number of publications per year were analysed, as well as publications per research area, research centres and journal characteristics. RESULTS: Health research production in Colombia is mainly published in international journals. Most published papers are linked to basic biomedical research overshadowing other areas of health research such as clinical or public health research. Universities are the main entities producing knowledge re health and their participation has increased during the last few years. CONCLUSIONS: Colombian health research production has increased during the last few years as shown by the increased numbers of Colombian papers in MEDLINE and LILACS during the period being studied. However, this production is still lower than that of other Latin American countries having similar socioeconomic and demographic conditions. PMID- 16703961 TI - [Social capital, violent deaths, and cancer mortality in Colombia: a population approach]. AB - OBJECTIVE: In Colombia there are evidences that social capital (SC) is associated with greater rates of violent crime ("perverse" SC). This study explores the relation between SC, violent deaths (1973-1996), and the accumulated occurrence of cancer deaths (1990-1996). MATERIAL AND METHODS: An ecologic study with the 33 Colombian departments was carried out. Correlations between violent deaths (inverse proxy of SC), the internal displacement ratio, and the mortality rates by each type of cancer. With robust regressions the effect of violent deaths ("perverse" SC) on the occurrence of mortality cancer were explored, adjusting by economic convergence or polarization (1960-1995), and the internal displacement ratio until 1996. RESULTS: Positive correlations (p < 0.05) between violent deaths ("perverse "CS) and all types of cancer, except breast and lung among men, were observed. In all the cases statistically significant associations were observed, after adjust by departments' economic convergence/polarization and internal displacement. DISCUSSION: This study shows a direct relation between violent deaths on the occurrence of cancer. The findings obtained here suggest an inverted U shape relation between SC and disease occurrence. PMID- 16703962 TI - [A scale for measuring the quality of post-anaesthetic recovery from the user's point of view]. AB - OBJECTIVE: Developing a valid scale for measuring the quality of post-anaesthetic recovery (QoR) from the patients' point of view. METHODS: ASA I or II scheduled surgical patients were selected. SETTING: Hospital San Juan de Dios, Bogota, Profamilia's Sexual and Reproductive Health Clinic and Social Security's Clinica Carlos Lleras. Five phases: semistructured interviews for establishing different quality categories and items referred by patients (n=30); content analysis and categorisation to establish relevant items (n=42); facial validity (n=20); scale design and validation, QoR (n=283): main factor analysis, varimax rotation Cronbach's alpha coefficient was used for testing internal consistency. Concurrent and discriminating criteria were validated by using non-parametrical statistics]; and reproducibility (n=100), tested by Kendal's concordance coefficient. RESULTS: The QoR was constructed in Colombian Spanish with 14 items and 3 domains, general quality (8 items), recovery room (5 items) and pain (one item). The QoR showed good internal consistency (Cronbach = 0.8783). The QoR positively correlated (Spearman's rho > 0.39) with 3 out of 5 scales and found differences by gender, kind of surgery, surgical site and kind of anaesthesia (Ji2 and K. Wallis, p < 0.05). The QoR had good reproducibility (Kendal = 0.6378, p = 0.0335). CONCLUSION: The QoR is the first valid scale constructed from the patient's point of view for measuring the quality of post-anaesthetic recovery. PMID- 16703964 TI - [Validating the Simpson-Angus Extrapyramidal Collateral Symptom Evaluation Scale]. AB - OBJECTIVES: The study was aimed at validating the Simpson-Angus scale for neuroleptic-induced extrapyramidal syndrome in Colombia, using a Spanish version of the scale. METHODS: The scale was validated in four steps: translating the scale, pilot study, preliminary use for factorial analysis and analysis of validity, reliability and sensitivity to change. RESULTS: Factorial structure and internal consistency were evaluated in 86 psychiatric inpatients. Test-retest and inter-rater reliability were evaluated in a 15-patient subgroup. Concurrent validity was analysed by simultaneously applying Simpson-Angus and Chouinard scales. Sensitivity to change was evaluated by comparing the scores of two different clinical points in a 20-patient subgroup. Main factor analysis revealed that the scale has a one-dimensional structure; the Glabella tap did not contribute towards total variability of the condition measured for the scale. Test-retest and inter-rater reliability values, concurrent validity and sensitivity to change were good. Correlation estimation scores were above 0.8 and accuracy levels were also good. CONCLUSIONS: The Spanish version of the SA scale, adapted for this study, is suitable for assessing neuroleptic-induced extrapyramidal syndrome; it would be a useful instrument in both clinical practice and research settings in Colombia. However, an important limitation could be its lack of detecting any domain different to that of rigidity. PMID- 16703963 TI - [Emotional burn-out in official teachers, Medellin, Colombia, 2005]. AB - OBJECTIVE: Exploring signs of emotional and physical weakness or Burnout syndrome in qualified teachers in Medellin, Colombia, 2005. METHODS: A cross-sectional study was applied to a random bi-stage sample of 239 teachers who filled in a questionnaire by themselves to explore demographic, work and social variables and dimensions of the Burnout syndrome, according to the Maslach Burnout Inventary (MBI). RESULTS: The findings showed that 23.4% of the sample presented signs of Burnout and a further 23.4% were at risk of suffering it; emotional weakness and depersonalisation were present in both subgroups. CONCLUSIONS: A relationship between Burnout signs and some of the variables studied was found. It is suggested that personal, family and social variables potentially associated with the syndrome should be explored more deeply. PMID- 16703966 TI - [The prevalence of nicotine-dependency in some populations: a systematic review]. AB - Tobacco use is the leading cause of preventable morbidity-mortality around the world. Nicotine dependence (ND) explains why people continue to smoke even though the harmful outcome associated with tobacco use is well-known. The object of this review was to establish current and one-year prevalence of ND among adults from the general population. Ebsco, Embase, Lilacs, Ovid, Proquest, and PubMed were systematically reviewed. The key words used were: smoking, tobacco, cigarette, nicotine, dependence, adults, general population and cross-sectional study. Only research carried out in English, Portuguese and Spanish was reviewed. A descriptive analysis of six articles was made. Current ND prevalence ranged from 27.8% to 55% (44.7% weighted average) and annual prevalence from 26% to 46.9% (37.7% weighted average). ND was independent of gender and higher amongst heavy smokers. It was concluded that ND prevalence is close to 45% amongst current regular smokers and affected 38% of smokers during the last year. More research is needed. PMID- 16703965 TI - [Diarrhoea-causing agents in children aged less than five in Tunja, Colombia]. AB - OBJECTIVES: Determining the prevalence and type of infectious ADD-causing agents in a sample of children aged less than five who consulted the IPS in Tunja during 2004. MATERIAL AND METHODS: A cross-sectional study was designed. Data was obtained by surveying 129 children younger than 5 suffering from ADD. Samples of faeces were obtained following outpatient consultation at San Rafael Hospital and SaludCoop's clinic in Tunja. RESULTS: Rotavirus was found in 48.1% of cases, Shigella in 0.8%, E. coli in 13.9%, Campylobacter in 2.3%, Giardia lamblia in 12.4% and E. hitolytica in 7%. The causative agent could not be identified in 15.5% of cases. The statistical association grew with age for Rotavirus (p < 0.01), E. coli (p < 0.05) and campylobacter (p < 0.001). CONCLUSION: Rotavirus is the major causative agent of ADD in children younger than one year and, generally, in children aged less than five. The prevalence found was similar to data reported in studies carried out in Facatatativa, Bogota, Santander, Manizales and the Choco in Colombia and studies carried out in Venezuela, Peru and Mexico. PMID- 16703968 TI - The evolution of the journal "Pain Physician". PMID- 16703967 TI - [Carbon monoxide contamination: an environmental health problem]. AB - Carbon monoxide is considered to be a major factor contaminating earth's atmosphere. The main sources producing this contamination are cars using gasoline or diesel fuel and industrial processes using carbon compounds; these two are responsible for 80% of carbon monoxide being emitted to the atmosphere. This substance has a well-known toxic effect on human beings and its acute poisonous effects (including death) have been widely studied; however, its long-term chronic effects are still not known. During the last few years, experimental research on animals and studies of human epidemiology have established the relationship between chronic exposure to low and middle levels of carbon monoxide in breathable air and adverse effects on human health, especially on organs consuming large amounts of oxygen such as the heart and brain. Harmful cardiovascular and neuropsychological effects have been documented in carbon monoxide concentration in air of less than 25 ppm and in carboxyhaemoglobin levels in blood of less than 10%. The main cardiac damage described to date has been high blood pressure, cardiac arrhythm and electrocardiograph signs of ischemia. Lack of memory, attention, concentration and Parkinson-type altered movement are the neuropsychological changes most frequently associated with chronic exposure to low levels of carbon monoxide and carboxyhaemoglobin. PMID- 16703969 TI - Evaluation of therapeutic thoracic medial branch block effectiveness in chronic thoracic pain: a prospective outcome study with minimum 1-year follow up. AB - BACKGROUND: The prevalence of persistent upper back and mid back pain due to involvement of thoracic facet joints has been described in controlled studies as varying from 43% to 48% based on IASP criteria. Therapeutic intraventions utilized in managing chronic neck pain and low back pain of facet joint origin include intraarticular injections, medial branch nerve blocks, and neurolysis of medial branch nerves by means of radiofrequency. These interventions have not been evaluated in managing chronic thoracic pain of facet joint origin. OBJECTIVE: To determine the clinical effectiveness of therapeutic thoracic medial branch blocks in managing chronic upper back and mid back pain of facet joint origin. DESIGN: A prospective outcome study. SETTING: Interventional pain management setting in the United States. METHODS: Fifty-five consecutive patients meeting the diagnostic criteria of thoracic facet joint pain by means of comparative, controlled diagnostic blocks were included in this evaluation. All medial branch blocks were performed in a sterile operating room under fluoroscopic visualization with mild sedation with midazolam and/or fentanyl. Statistical methods incorporated intent-to-treat analysis. OUTCOME MEASURES: Numeric pain scores, significant pain relief > or = 50%), Oswestry Disability Index, work status and Pain Patient Profile (P-3). Significant pain relief was defined as an average 50% or greater reduction of numeric pain rating scores. RESULTS: The results showed significant differences in numeric pain scores and significant pain relief (50% or greater) in 71% of the patients at three months and six months, 76% at 12 months, 71% at 24 months, and 69% at 36 months, compared to baseline measurements. Functional improvement was demonstrated at one year, two years, and three years from baseline. There was significant improvement with increase in employment among the patients eligible for employment (employed and unemployed) from baseline to one year, two years, and three years (61% vs 96% to 100%) and improved psychological functioning. CONCLUSION: Therapeutic thoracic medial branch blocks were an effective modality of treatment in managing chronic thoracic pain secondary to facet joint involvement confirmed by controlled, comparative local anesthetic blocks. PMID- 16703970 TI - Quality assurance for interventional pain management procedures. AB - BACKGROUND: Over the last decade various guidelines for quality assurance in pain medicine have been published for cancer pain, acute postoperative pain and other pain conditions. However, quality assurance for interventional pain management procedures has yet to be addressed. An interventional pain program should at least evaluate 1) efficacy of pain relief; 2) complication rate; and 3) patient satisfaction. OBJECTIVE: This study was designed to monitor the quality of interventional pain management procedures in a university teaching hospital. STUDY DESIGN: A prospective survey. METHODS: From January 1, 2004, to June 30, 2004, the quality of interventional pain management procedures in a university teaching hospital in Miami, Florida was monitored. Questionnaires assessing immediate pain relief, patient satisfaction, and complications were provided to each patient and physician immediately after completion of each procedure. Data was collected before patients were discharged. RESULTS: A total of 566 patients with a mean age of 52.9 years participated in the survey. Interventional pain management procedures included epidural steroid injections, facet joint blocks, transforaminal epidural injections, sympathetic nerve blocks, lumbar discography, nucleoplasty, percutaneous disc decompression, spinal cord stimulator trial, and intravenous regional blocks, etc. Among 528 patients who reported their pain scores before and after procedures, 487 (92%) patients reported various degrees of pain relief immediately following their procedures. The average pain score decreased 4.7 on a o to 10 scale after treatment (p < 0.001). No major complications were reported for this group of patients. Among 442 patients who answered the question regarding satisfaction, 406 (91.8%) were satisfied, or highly satisfied, with the immediate outcome of their procedures. CONCLUSION: The results of the current study indicate that quality assurance of interventional pain management procedures in terms of immediate pain relief following the procedure, low complication rate, and high patient satisfaction can be achieved through application of a quality assurance program. PMID- 16703971 TI - Treatment of painful advanced internal lumbar disc derangement with intradiscal injection of hypertonic dextrose. AB - BACKGROUND: Degenerative discs are thought to produce nerve root pain either mechanically or chemically. Particularly in the case of advanced degenerative disc disease, this clinical entity has often proven to be symptomatically resistant to peridural steroids, Intra-discal Electrothermoplasty (IDET) and direct surgical intervention. Exposure of irritated nerves to hypertonic dextrose is thought to have chemoneuromodulatory potential. Sustained pain reduction has been demonstrated in a pilot study involving injection of a combination of dextrose, glucosamine, chondroitin and dimethylsulfoxide into degenerative discs of patients with chronic low back pain of discogenic origin. OBJECTIVE: To assess the effects of disc space injections of hypertonic dextrose in patients experiencing chronic advanced degenerative discogenic leg pain, with or without low back pain. STUDY DESIGN: Prospective consecutive patient series. METHODS: Patients with moderate to severe degenerative disc disease without herniation and with concordant pain reproduction with CT discography were included. All had failure of a physical therapy trial and substantial but temporary relief with two fluoroscopically guided epidural steroid injections. Patients underwent bi-weekly disc space injection of a solution consisting of 50% dextrose and 0.25% Bupivacaine in the disc(s) found positive on discography. The study was performed in an out-patient surgery center in the United States. Outcome measures included an 11-scale numeric pain score (0-10). RESULTS: Each patient was injected an average of 3.5 times. Overall, 43.4% of patients fell into the sustained improvement group with an average improvement in numeric pain scores of 71%, comparing pretreatment and 18 month measurements. CONCLUSION: The results suggest that intradiscal injection of hypertonic dextrose may have a place in the management of pain arising from advanced lumbar degenerative disc disease. PMID- 16703972 TI - Does random urine drug testing reduce illicit drug use in chronic pain patients receiving opioids? AB - BACKGROUND: Prescription drug abuse and illicit drug use are common in chronic pain patients. Adherence monitoring with screening tests, and urine drug testing, periodic monitoring with prescription monitoring programs, has become a common practice in recent years. Random drug testing for appropriate use of opioids and use of illicit drugs is often used in pain management practices. Thus, it is expected that random urine drug testing will deter use of illicit drugs, and also improve compliance. OBJECTIVES: To study the prevalence of illicit drug use in patients receiving opioids for chronic pain management and to compare the results of illicit drug use with the results from a previous study. DESIGN: A prospective, consecutive study. SETTING: Interventional pain management practice setting in the United States. METHODS: A total of 500 consecutive patients on opioids, considered to be receiving stable doses of opioids supplemental to their interventional techniques, were studied by random drug testing. Testing was performed by rapid drug screen. Results were considered positive if one or more of the monitored illicit drugs including cocaine, marijuana (THC), methamphetamine or amphetamines were present. RESULTS: Illicit drug use was evident in 80 patients, or 16%, with marijuana in 11%, cocaine in 5%, and methamphetamine and/or amphetamines in 2%. When compared with previous data, the overall illicit drug use was significantly less. Illicit drug use in elderly patients was absent. CONCLUSION: The prevalence of illicit drug abuse in patients with chronic pain receiving opioids continues to be a common occurence. This study showed significant reductions in overall illicit drug use with adherence monitoring combined with random urine drug testing. PMID- 16703973 TI - A novel approach to transnasal sphenopalatine ganglion injection. AB - BACKGROUND: Sphenopalatine ganglion block is used to treat headache and facial pain. There are transnasal, transoral and lateral approaches to establish the blockade. The transnasal application of topical anesthetic is the simplest and most common technique among the three. However, the diffusion of topical anesthetic to the ganglion is unpredictable and the blockade is not durable. OBJECTIVE: To describe a novel transnasal injection technique to block sphenopalatine ganglion. METHODS: The technique includes topical anesthesia to the nasal mucosa between the middle and inferior turbinates from nares to the nasopharynx, followed by a needle insertion in the same tract guided by its tailored plastic cover-sheath for blocking agent injection to the ganglion. RESULTS: Fluoroscopy revealed that with this technique the needle tip located near the medial pterygoid plate at the level of the middle concha and contrast flowed superoposteriorly to the maxilla without vascular uptake. Pain relief for this patient was prolonged from one day with topical technique to four weeks with this technique. The subsequent blocks without fluoroscopy provided similar prolonged pain relief. CONCLUSION: This method preserves the technical simplicity of the original transnasal approach and includes the predictability and durability of the blockade from needle injection. PMID- 16703974 TI - A comparison of pulsed radiofrequency and continuous radiofrequency on thermocoagulation of egg white in vitro. AB - BACKGROUND: Clinical studies have demonstrated the efficacy of pulsed radiofrequency (PRF). PRF energy is delivered to neural structures via specifically designed, percutaneously placed needles to treat some chronic pain states. PRF was introduced as a non-destructive alternative to destructive lesioning produced by continuous radiofrequency (CRF) energy. However, there is an ongoing controversy regarding the potential tissue-destructive effects of PRF used for pain management. OBJECTIVE: To evaluate the ability of PRF to coagulate egg white at various temperatures used clinically and to compare with CRF. METHODS: A commercially available (TYCO-Radionics Labs) 5 cm, 22G (0.7 mm) SMK needle with 5 mm active tip was inserted into a 10 mL test tube containing raw egg white at 37 degrees C and the tip was heated up to 80 degrees C. The photographic patterns of thermocoagulation of egg white in vitro produced by continuous and pulsed radiofrequency (RF) were compared and the lowest temperature at which PRF produced thermocoagulation was determined. RESULTS: Pulsed RF produced barely detectable thermocoagulation at 60 degrees C. Above 60 degrees C, the pattern of coagulation produced by PRF resembled that observed with CRF. However, the density and size of the coagulation ball appeared somewhat greater with CRF. CONCLUSION: PRF coagulated egg white at temperatures above 60degrees C in a manner similar to CRF. Monitoring needle tip temperature using the thermode supplied with the needle during PRF and keeping the recorded tip temperature below 60degrees C may minimize unwanted thermal destruction of tissue. PMID- 16703975 TI - Percutaneous lumbar disc decompression. AB - Chronic low back pain is a major social, economic, and healthcare issue in the United States. Various techniques are utilized in managing discogenic pain, with or without disc herniation. Percutaneous techniques are rapidly replacing traditional open surgery in operations requiring discectomy, decompression, and fusion. The percutaneous access to the disc was first used in the 1950s to biopsy the disc with needles. Percutaneous access to the disc using endoscopic techniques was developed in the 1970s. Technical advances in the use of intradiscal therapies led to the development of intradiscal electrothermal annuloplasty (IDET), DISC Nucleoplasty, and DeKompressor, along with laser assisted, endoscopic, and Nucleotome disc decompressions. The indications for percutaneous lumbar disc decompression include low back and lower extremity pain caused by a symptomatic disc. Internal disc disruptions and disc herniations are common causes of low back and/or lower extremity pain which may become chronic, if not diagnosed and treated. Annular tears lead to migration of the nuclear material and deranged internal architecture. In the chronically damaged intervertebral disc, leakage of nuclear material from annular tears can initiate, promote, and continue the inflammatory process and delay or stop recovery of vital remaining intradiscal tissue. The most often stated goal of central nuclear decompression is to lower the pressure in the nucleus and to allow room for the herniated fragment to implode inward. Provocative discography prior to percutaneous lumbar disc decompression is recommended. Percutaneous disc decompression may result in a small number of complications but occasionally, these could be serious. PMID- 16703976 TI - Treatment challenges and complications with ziconotide monotherapy in established pump patients. AB - BACKGROUND: The U.S. Food and Drug Administration (FDA) recently approved Ziconotide intrathecal infusion for the management of severe chronic pain in patients for whom intrathecal therapy is warranted, and who are intolerant of, or refractory to, other methods of treatment, including intrathecal morphine. Ziconotide is approved as a monotherapy, but there are challenges associated with the decision to wean intrathecal opioids for Ziconotide alone. Maintaining adequate analgesia and managing opioid withdrawal symptoms may be difficult. Additionally, a variety of adverse physiological, cognitive and psychiatric events may be associated with this new drug. Patients with pretreatment psychiatric disorders may be at increased risk for treatment complications. OBJECTIVE: To present a report of a case series describing treatment challenges and complications associated with the decision to convert established pump patients from intrathecal opioid therapy to Ziconotide monotherapy. DESCRIPTION OF CASES: Three established pump patients, refractory to intrathecal opioid therapy, were converted to Ziconotide monotherapy. All of these patients experienced significant emotional distress or psychological symptoms that threatened the success of the treatment. Achieving adequate analgesia, reducing Ziconotide to mitigate adverse physiological effects, managing opioid withdrawal symptoms, and supportive psychological consultation were combined to achieve successful outcomes in two of our three patients. CONCLUSION: This report describes challenges associated with the decision to convert established pump patients from intrathecal opioid therapy to Ziconotide monotherapy. Inadequate analgesia, adverse medication effects, and opioid withdrawal symptoms can precipitate a stressful situation that may be perceived as dangerous or threatening by patients who are predisposed to anxiety. Screening patients for psychiatric disorders, anxiety-proneness and/or vulnerability to stress should be considered to reduce the risk of treatment complications. A multimodal approach is strongly advocated, including rapid responses of treating physicians and nurses along with strong psychological support. PMID- 16703978 TI - A modification of the interscalene brachial plexus block technique that allows multiple blocks to be done with minimal morbidity and discomfort. PMID- 16703977 TI - Pulsed radiofrequency of lumbar nerve roots for treatment of chronic inguinal herniorraphy pain. AB - BACKGROUND: Inguinal hernia repairs are commonly performed and although not seen in a majority of patients, chronic inguinal pain can be a debilitating state resulting after inguinal hernia repairs. Treatment options exist, including pharmacological and surgical management, but with associated risks and side effects. METHODS: In this case series report, five patients with chronic inguinal pain were selected. After initial positive response to T12, L1 and L2 nerve root blocks, pulse radiofrequency (PRF) was performed. RESULTS: All patients reported 75% to 100% pain relief lasting from six to nine months. DISCUSSION: Several hypotheses have been proposed, including selective delta and c-nerve fiber destruction, upregulation of intermediate early gene expression (IEG) and increase in c-fos protein that modulated pain transmission. CONCLUSION: The mechanism of action of pulsed radiofrequency remains unclear. This case study demonstrates the effectiveness of minimally invasive neurodestruction of T12, L1 and L2 nerve roots utilizing Pulse Radiofrequency fields. PMID- 16703979 TI - How to obtain an SI Joint arthrogram 90% of the time in 30 seconds or less. PMID- 16703980 TI - 2004 National Hospital Discharge Survey. AB - OBJECTIVES: This report presents national estimates of the use of non-Federal short-stay hospitals in the United States during 2004 and selected trend data. Numbers and rates of discharges, diagnoses, and procedures are shown by age and sex. Average lengths of stay are presented for all discharges and for selected diagnostic categories by age and by sex. METHODS: The estimates are based on medical abstract data collected through the 2004 National Hospital Discharge Survey (NHDS). The survey has been conducted annually by the Centers for Disease Control and Prevention's National Center for Health Statistics (NCHS) since 1965. Diagnoses and procedures presented are coded using the International Classification of Diseases, 9th Revision, Clinical Modification, or ICD-9-CM. RESULTS: Trends in the utilization of non-Federal short-stay hospitals show that the rate of hospitalization of the elderly (those 65 years and over) increased 24 percent from 1970 through 2004 despite a decrease in the 1980s. The rates for the other age groups declined overall. In 2004, those 65 years and over comprised 12 percent of the U.S. population, 38 percent of all hospital discharges, and used 44 percent of all inpatient days of care. In 2004, there were an estimated 34.9 million hospital discharges, excluding newborn infants. The average length of stay was 4.8 days for all inpatients and 5.6 days for the elderly. Almost one half of hospital stays for heart disease had a first-listed discharge diagnosis of either congestive heart failure (25 percent) or coronary atherosclerosis (24 percent). There were 45 million procedures performed on inpatients during 2004. From 1995 through 2004, for those 65 years and over, the rate of hip replacements increased 38 percent, and the rate of knee replacements increased 70 percent. One quarter of all procedures performed on females were obstetrical. Almost one quarter of all procedures performed on males were cardiovascular. PMID- 16703981 TI - Court cases highlight the importance of clear nursing records. PMID- 16703983 TI - Are annual health MOTs a good idea? PMID- 16703984 TI - Are annual health MOTs a good idea? PMID- 16703985 TI - The next nursing century. PMID- 16703986 TI - Training for crisis resolution home treatment teams. AB - The changing training needs of the mental health workforce have been increasingly highlighted in recent years with the national development of new services like crisis resolution/home treatment teams. This article presents some key changes made to short training courses in crisis resolution. PMID- 16703987 TI - Sarcoma. PMID- 16703988 TI - Taking a midstream specimen of urine. AB - Collecting midstream specimens of urine (MSU) is a common nursing activity. It is indicated for adults and children who are continent and can empty their bladder on request. The general principle is to collect the midstream or middle part of a void if a urinary tract infection (UTI) is suspected. Since this is one of the most frequently occurring infections, an MSU may be required in many areas of clinical practice. Yet despite being a regular nursing activity, the procedure is often undertaken incorrectly. PMID- 16703989 TI - An alternative to surgery in treating ectopic pregnancy. AB - In an ectopic pregnancy the fertilised egg becomes implanted outside the uterus. It affects around one in every hundred pregnancies. Traditionally the condition has been managed surgically. However, in recent years many women have been treated with methotrexate therapy. PMID- 16703990 TI - Orlistat. PMID- 16703991 TI - Using post-take ward rounds to facilitate simple discharge. AB - This article discusses the evolution of simple discharge/transfer destination labels into a post-take ward round form. The aim was to improve the clarity of the management plan and discharge/ transfer decisions including an estimated length of stay on the post-take ward rounds taking place on the emergency admissions ward. PMID- 16703992 TI - Opportunities for nurses in a private finance initiative. AB - The private finance initiative is part of the government's building procurement policy, incorporating both public and private sectors. This article outlines the part nurses can play, as a clinical adviser, within a PFI project team and within particular aspects of the PFI process. PMID- 16703993 TI - Making your development portfolio work for you. PMID- 16704002 TI - [Value of corpitolinol 60 in geriatric decubitus prevention protocols. (GIPPS study)]. PMID- 16704003 TI - [A diet education adapted to the cultural habits of diabetic patients]. PMID- 16704004 TI - [Pinel, Pussin, an encounter]. PMID- 16704005 TI - [Dates and figures marking the nursing profession in psychiatry]. PMID- 16704006 TI - [Psychiatric nursing in search of status from the beginning of the last century]. PMID- 16704008 TI - [A testimonial to nursing care in mental health]. PMID- 16704007 TI - [Sister O., Mrs. E., the forgotten nurses in the psychiatric revolution]. PMID- 16704009 TI - [Nursing interview in the management of care]. PMID- 16704010 TI - [Jean-Louis Gerard, master champion of psychiatric nurses]. PMID- 16704011 TI - [A communal history to construct]. PMID- 16704012 TI - [Education in psychiatry, and tomorrow?]. PMID- 16704013 TI - [Bibliography. Psychiatric nursing care, 200 years of history]. PMID- 16704014 TI - [3/10 Undesirable side-effects of drugs: causes, consequences, prevention]. PMID- 16704015 TI - [2/4 Level 1 analgesics]. PMID- 16704016 TI - [Loneliness intervention in the aged: regulation and efficacy]. PMID- 16704017 TI - [Experience of family members with the decision concerning artificial nutrition and hydration in people with dementia in nursing homes]. AB - The purpose of this study is to investigate how relatives of nursing home patients with dementia, for whom the decision whether to start or forgo artificial nutrition and hydration (ANH) was made, assess the decision-making process. Furthermore we evaluated the information given by the nursing home staff to the relatives and the care provided by them to the patient. The study was an observational study based on written questionnaires. Relatives of 99 nursing home patients with dementia filled in a questionnaire at the time of the decision making and 6 weeks after the decision was made or 6 weeks after the patient had died. Almost all relatives assessed the 4 aspects of the decision-making process (number of consults, content of the consults, carefulness of the decision-making and there own part in the decision-making process) at both times as 'good' or 'satisfactory'. Furthermore it appeared that they judged the carefulness of the decision-making process significantly more often as 'good' when they also assessed the other three aspects as 'good'. Especially their own influence on the decision-making process appeared to play a part in this matter. Information about the possible consequences of starting ANH as well as about the condition of the patient were experienced sufficient by almost all relatives. This was also the case for the care offered to the patient. In general almost all relatives reported satisfaction with the decision and with the carefulness of the decision making process. This satisfaction did not differ between the moment of the decision and afterwards. For this it did not make any difference whether the patient had or had not died. PMID- 16704018 TI - [Construct validity of some episodic memory tests for psychogeriatric patients]. AB - Episodic memory is the conscious recollection of personal experiences. In clinical practice several episodic memory tests are used, but their validity as measures of an episodic memory construct is not clear. The sensitivity and specificity of variables hypothesized to represent a distinct psychological construct is established by a pattern of convergent and discriminant validity. Confirmatory factor analysis was used to analyse the correlations between four episodic memory tests: a standardized orientation questionnaire, free recall, and two delayed recognition memory tests. The episodic memory construct was contrasted with an executive functioning construct, measured by three variables: two tasks of semantic word fluency and the Expanded Mental Control Test. The measures were taken from 813 consecutive visitors of a psychogeriatric day care centre. As a set the four indicators of episodic memory provided reliable measurement of the construct. The same was true for the three measures of the executive functioning construct. However, the strong correlation (0.89) between the two constructs implies a lack of discriminant validity and suggests that processes of executive control contribute to the successful performance on episodic memory tasks. PMID- 16704019 TI - [Elderly heart failure patients and the role of beta-blocker therapy]. AB - In this article different aspects of chronic heart failure in old age are described. We mainly focus on the place of beta-blocker therapy in chronic heart failure. Beta-blockers are recommended for the treatment of stable chronic heart failure with left ventricular systolic dysfunction. There is additional information from recent studies that there is proven efficacy for beta-blocker therapy in patients with heart failure up to the age of 80 years. For patients with heart failure aged 80 and over the evidence to prescribe beta-blockers is limited. However, it is known that also in very elderly patients beta-blocker therapy is well tolerated. In patients with heart failure with preserved systolic ventricular function there is still no evidence that there is a beneficial effect of beta-blockers. It is still not clear if there are differences between beta blocking agents. Of all beta-blockers, only bisoprolol, carvedilol, nebivolol and metoprolol CR are proven effective in stable chronic heart failure with impaired left ventricular systolic function and can be recommended in elderly patients on standard treatment with diuretics and ACE inhibition. PMID- 16704020 TI - Prevalence of post-treatment pain after cleaning and shaping of the root canal system using manual step-back versus rotary nickel titanium. AB - To evaluate the effect of ProTaper rotary NiTi instrumentation technique on post treatment pain at different time periods when compared with step-back technique, one hundred forty-six patients with 160 teeth participated in this study. All treated teeth (160) are having vital pulps and divided into two groups according to the canal instrumentation technique. Group (1) consisted of 80 teeth, cleaned and shaped with manual step-back technique using K-Flexo hand files. Group (2) was 80 teeth cleaned and shaped using ProTaper (NiTi) rotary files. Patients assessed post-treatment pain at 8 h, 24 h, and 48 h as none, mild, moderate, or severe. Evaluation of the two groups, according to the three post-treatment time period, was tabled as to the number of responses at each pain level at each interval. Comparisons were made by subjecting the collected data to a multiple- regression test. The results demonstrated that there were no statistically significant difference in post-treatment pain between the two groups at any time period P > 0.05. PMID- 16704021 TI - [Mouth diseases and antibiotic therapy practices in Yaounde]. AB - Based on 309 patients (171 women and 138 men), consulted by 15 dental surgeons of Yaounde during one year, the aim of this study was the evaluation of bringing into use an antibiotherapy in daily practice. A questionnaire on the subject was distributed to practitioners. We registered 10 most frequent mouth dental diseases that needed curative antibiotherapy, being 87% of prescriptions. A prophylactic antibiotherapy has been prescripted in three per cent of cases to patients carriers of general pathologies (cardiopathies, diabete...) or mandibular fractures. In 10% of cases, a covering antibiotherapy has been established during simple extractions of teeth without infections, from healthy patients. In 48% of cases the duration of antibiotherapy was short (less than eight days) and in 32% of cases, the quantities were infratherapeutic. There were three per cent of prescriptions concerning antibiotics (sulfonamides, phenicoles...) unsuited to usual germs of mouth cavity. Generics drugs have been prescribed in 12% of cases. The more prescribed antibiotics were amoxicillins, macrolides and cephalosporines. Only a best knowledge of antibiotics' families and their indications are able to avoid clumsiness. At last generic drugs prescription in our difficult economical environment should be common. PMID- 16704022 TI - Weaning practices among some Nigerian women: implication on oral health. AB - The objective of this study was to document the various weaning diets given by some Nigerian women and consider their implication on the oral health of their children. A cross sectional survey was conducted among 700 volunteer mothers of young children aged between 6-18 months in Ibadan, a city in south-western Nigerian. Results revealed that pap was the most frequently given food to the infants as 607 (86.7%) of them ate it. Chocolate beverages, natural fruit juice and soft drinks were the most commonly given drinks as 88.1%, 79.9% and 70.3% babies were said to be given these respectively. Over 57% of mothers sweetened pap with sugar. Forty seven percent of mothers added glucose to children's drinking water. Over two-third (64.9%) of children ate biscuits several times a day. Soft drinks, commercial fruit juices and squash were consumed by 16.1%, 9.6% and 7.7% of the infants respectively on a daily basis. Judging by the findings of this study, it is evident that many infants are being exposed to a highly cariogenic diet at an early age. In the light of the harmful effect of these foods and drinks on the body and teeth, it is important that oral health care workers initiate moves to discourage this habit via health education programmes. Nurses, community health workers at various levels, pregnant women and mothers of young children should be the targets of such intervention. PMID- 16704023 TI - [Cellulitis and phlegmons of dental origin in the CHU of Yaounde]. AB - Cellulitis and phlegmons of dental origin in the CHU of Yaounde. Cellulitis and phlegmons are frequent complications of teeth infections in tropics in general and particularly in Cameroon. They concern all ages and affect more men than women. These pathologies are more developed in disfavoured social classes. Their frequency is very high among pupils and students, whose the financial power depends on poor parents. Cellulitis and phlegmons are more induced by molar's infections. The upper and lower parts of the check were the most frequent. These pathologies disturb the life because of pain they entertained and they can modify life prognosis. They constitute emergencies for diagnosis and treatment. PMID- 16704024 TI - [Study of tooth shade in Black Africans]. AB - The 243 sample prospective study shows specifities about black African teeth shade particularly the colour. The results after the use of a study mixed shader composed from three standard shaders are; first: the sequence from the clearest tooth to the less is: central incisor, lateral incisor. Second: the maxillary teeth shade is clearer than the mandibulars. Third, 16% teeth shade not be conducted to determine the objective parameters for black African dentogenics. Therefore: esthetic restorative treatment will be more efficient. PMID- 16704026 TI - Schisms in the tobacco control movement (Part 2). PMID- 16704025 TI - [Maxillo-facial prostheses: an issue in public health]. AB - BACKGROUND: Acquired or congenital loss of structure can cause facial deformity. This can destroy identity and lead to certain forms of exclusion. Surgical and sometimes prosthetic reconstruction is required. The Maxillofacial Prosthesis (P.M.F) can be defined as the art and science of artificial reconstruction of facial bones. There is a continual increase in patient demand for this type of prosthesis with the development of new materials, methodologies and techniques. METHODS: In Toulouse, the Rangueil University Hospital houses a maxillofacial prosthesis consultation facility linked to the Odontology Service. An exhaustive descriptive study carried out retrospectively on the basis of the files of all patients examined between July 1996 and July 2002 will provide us with an understanding of patient characteristics and also enable us to quantify the level of satisfaction of requirements for rehabilitation of these losses of maxillofacial structure. RESULTS: 215 patient files were processed between July 1996 and July 2002. Separating the patients by sex shows a male prevalence of 66%. An analysis of variations between male and female patients for different aetiologies shows that tumour pathologies are more common in men and that this distinction is all the more marked as regards traumatic facial injuries and in particular suicide attempts. In total, 401 maxillofacial prostheses were carried out, including 116 facial epitheses (new prostheses or renewal of prosthesis). CONCLUSION: Loss of facial structure, whatever its aetiology constitutes a real public health issue. The maxillofacial prosthesis will allow cancer monitoring of the site while at the same time offering out patient treatment. Above and beyond its crafted or empirical aspect, it should find its place in medicine thanks to the overall scope of the rehabilitation that it can offer from both an aesthetic point of view but also a the psychological one, but above all from a functional perspective. PMID- 16704027 TI - Management of chronic asthma in adults in diverse regions of the world. AB - Asthma is a chronic inflammatory disease of the lungs associated with significant morbidity and mortality worldwide. Adoption of current treatment guidelines that propose inhaled corticosteroids (ICS) as the foundation for asthma treatment should control most patients with chronic asthma. Rapid-acting inhaled beta (beta) 2-agonists are best reserved for acute symptom relief. Long-acting beta-2 agonists in combination with ICS are the most effective asthma treatment currently available when asthma is not controlled on low-dose ICS alone; however, they are not universally available due to cost. Slow-release theophylline may be an alternative cost-effective add-on therapy to ICS in resource-poor areas, although its potential for toxicity has limited its use over the last decade. New targeted anti-inflammatory therapies lack the broad anti-inflammatory activity of ICS and are unaffordable in most settings. Implementation of guidelines for asthma care is an unresolved challenge, and major gaps in asthma care are consistent across the globe. Review of asthma management worldwide shows that control of the disease in relation to the Global Initiative for Asthma (GINA) goals of asthma treatment is not achieved in a large proportion of patients, despite the widespread availability of guidelines and even with access to effective treatment in resource-rich settings. Many resource-poor countries have the additional challenge of lack of access to basic asthma treatment such as ICS. The challenge is to provide global access to core asthma medications, particularly ICS, at affordable prices, to improve implementation of treatment guidelines and to encourage better health care provider and patient education. PMID- 16704028 TI - Managing acute attacks of asthma. AB - Asthma patients may present to the health services with acute attacks. Assessment and management must be undertaken rapidly. The patient must be given oxygen and then assessed by measuring the peak expiratory flow rate (PEF) before and after salbutamol, and by questioning and examining the patient. Using the information gathered, the caregiver is able to assess the grade of severity of the attack and provide appropriate care according to the severity. Patients judged as having imminent respiratory arrest must be immediately transferred to intensive care after starting treatment. Patients judged as having a severe attack are given oxygen, salbutamol, systemic corticosteroids and are closely monitored. They must remain for a minimum of 6 h prior to being either hospitalised or sent home. Patients with moderate attacks are given salbutamol and oral prednisone and are kept under observation for a minimum of 2 h. If stable at least 1 h after last dose of salbutamol, they may be sent home. Patients with mild attacks are given inhaled salbutamol and kept under observation for a minimum of 2 h. If stable at least 1 h after the last dose of salbutamol, they may be sent home. Careful and correct follow-up after an attack is crucial. PMID- 16704029 TI - Obstructive airways diseases, smoking and use of inhaled corticosteroids in southern Sweden in 1992 and 2000. AB - OBJECTIVE: To examine the prevalence of obstructive pulmonary diseases, respiratory symptoms, smoking habits and pulmonary medication in an adult population, and to compare the results with a study performed in the same geographical area in 1992. DESIGN: In 2000, a postal questionnaire was sent to a randomly selected population of 5179 subjects aged 20-59 years living in southern Sweden. RESULTS: The participation rate was 71.3%. Self-reported asthma was reported by 8.5% of all respondents (vs. 5.5% in 1992, P < 0.001) and 14.5% of females aged 20-29 years. Self-reported chronic bronchitis and/or emphysema and/or chronic obstructive pulmonary disease (CBE/COPD) was reported by 3.6% (vs. 4.6% in 1992, non-significant) with the highest prevalence (5.7%) in the 50-59 year cohort. Smoking decreased from 33.3% in 1992 to 28.4% in 2000 (P < 0.05). About 46% of asthmatics reported nocturnal respiratory symptoms, and 69% reported having had asthma symptoms in the last 12 months. Use of inhaled steroids increased in subjects with asthma and CBE/COPD from 19.4% to 36.5% (P < 0.05) and from 8.6% to 30.0% (P < 0.05), respectively. CONCLUSIONS: Self-reported asthma increased significantly between 1992 and 2000, but the prevalence of CBE/COPD was unchanged. The high proportion of reported symptoms in asthmatics despite an increased use of steroids suggests that further efforts are needed to improve asthma treatment. PMID- 16704030 TI - No gender difference in effectiveness of smoking cessation treatment in a Brazilian real-life setting. AB - BACKGROUND: The clinical effectiveness of pharmacotherapy for smoking cessation in real-life settings has yet to be evaluated. OBJECTIVES: To assess the effectiveness of bupropion in general clinical practice for smoking cessation and to identify predictors of failure. METHODS: In an open, non-randomised study, smokers were recruited at the Smoking Cessation Clinics, Hospital Sao Lucas, Porto Alegre, Brazil. Subjects participated in a motivational group meeting, completed a standardised questionnaire and Fagerstrom test, and had their vital signs and exhaled CO registered. All participants received a prescription of bupropion and the same cognitive behaviour therapy. They attended eight weekly individual sessions, then monthly until the sixth month and a final session at month 12. The primary outcome measure was the rate of abstinence at 12 months. The predictor factors studied were sex, age, educational level, nicotine dependence, previous attempts and comorbidities. RESULTS: Among 253 smokers (62.5% females), abstinence rates at 6 months were 20.8% for males and 22.7% for females. The success rates dropped to 13.9% and 14.3% for males and females, respectively. CONCLUSIONS: Cognitive therapy plus bupropion for smoking cessation in real-life clinics in Brazil were similar to the efficacy found in clinical trials. No significant gender differences in success rates were found. PMID- 16704031 TI - Lack of association between COPD and transforming growth factor-beta1 (TGFB1) genetic polymorphisms in Koreans. AB - OBJECTIVE: Many genetic variations have been suggested as genetic risk factors for the development of chronic obstructive pulmonary disease (COPD), including single nucleotide polymorphisms in the transforming growth factor-beta1 (TGFB1) gene. We attempted to elucidate the association between TGFB1 genetic polymorphisms and COPD among Koreans. DESIGN: The genotypes of 102 male patients with COPD and 159 volunteers with similar distributions of age, sex and smoking intensity, as well as normal pulmonary function, were determined for three previously associated TGFB1 single nucleotide polymorphisms (SNPs), -10807G/A (rs2241712) and -509T/C (rs1800469), located in or near the promoter, and 29T/C (rs1982073), located in exon 1 of the TGFB1 gene. RESULTS: No significant associations between COPD and the three TGFB1 SNPs could be identified. In addition, the haplotypes composed of three TGFB1 SNPs were not associated with the presence of COPD. CONCLUSION: These results differ from previous reports involving Caucasians, and might reflect racial differences in the pathogenesis of COPD. PMID- 16704032 TI - Serum procalcitonin distinguishes CAP due to bacteria, Mycobacterium tuberculosis and PJP. AB - SETTING: Procalcitonin (PCT), a propeptide of the hormone calcitonin, is a novel marker of the inflammatory response to infection. It has been used to discriminate between infectious and non-infectious causes of inflammation, and as a marker of severe sepsis in the intensive care unit. OBJECTIVE: To evaluate the utility of PCT in distinguishing community-acquired pneumonia (CAP) due to common bacteria, Mycobacterium tuberculosis and Pneumocystis jirovecii in a high human immunodeficiency virus (HIV) prevalence setting. METHODS: Two hundred and sixty six patients admitted with a diagnosis of CAP were investigated. Serum samples for PCT were collected on admission. PCT levels were measured using a commercial immunoluminometric assay. RESULTS: A microbiological diagnosis was obtained in 169/266 patients: 44 pulmonary tuberculosis (PTB), 31 P. jirovecii pneumonia (PJP), and 35 bacterial pneumonia. The PCT levels were PTB 4.16 ng/ml (SEM 1.197; 95% CI 1.749-6.579); PJP 1.138 ng/ml (SEM 0.2911; 95% CI 0.543-1.734); and bacterial pneumonia 19.48 ng/ml (SEM 5.64; 95% CI 8.021-30.938, P < 0.0004). Thirty-six had co-infections. CONCLUSION: PCT levels differ significantly in patients with CAP due to TB, PJP and bacteria. PCT may be important in distinguishing M. tuberculosis and PJP in a high HIV prevalence setting where atypical presentations often confound the empirical clinical diagnosis. PMID- 16704033 TI - Clinical presentation among HIV-infected and non-infected adults with community acquired pneumonia in Nairobi. AB - SETTING: Risk factors for mortality in hospitalized patients with community acquired pneumonia (CAP) are well known. There are limited data on prognostic indicators among out-patients. OBJECTIVE: To compare the clinical presentation, outcome and prognostic factors for clinical improvement in human immunodeficiency virus (HIV) infected and non-HIV-infected out-patients with CAP. METHODOLOGY: Adults in Nairobi with CAP were treated with erythromycin as first-line therapy. Clinical symptoms were evaluated using a validated CAP-related symptom score (CSS). Clinical improvement was defined as reduction of baseline CSS by > or = 50%. RESULTS: Of 531 adults enrolled with CAP, 422 (79.5%) completed follow-up. Participants had a mean age (+/- SD) of 33.7 +/- 11.4 years, 274 (51.6%) were male and 193 (37%) were HIV-seropositive with a higher baseline CSS (27 vs. 25, P < 0.006). Overall, 196 of 422 (46%) had clinical improvement by 28 days. Factors independently associated with a longer time to clinical improvement included not being married (adjusted hazard ratio [aHR] 0.66, 95% CI 0.48-0.92) and higher baseline CSS (aHR 1.05, 95% CI 1.03-1.06). CONCLUSIONS: HIV-infected and non infected patients with CAP responded similarly to out-patient treatment, but HIV infected patients were more likely to present with severe symptoms. Baseline CSS and marital status were predictive of time to clinical improvement. PMID- 16704034 TI - Screening for tuberculosis prior to isoniazid preventive therapy among HIV infected gold miners in South Africa. AB - SETTING: Human immunodeficiency virus (HIV) clinic for employees of a gold mine, Free State, South Africa. OBJECTIVE: To evaluate the process of screening for active tuberculosis (TB) prior to commencing TB preventive therapy in HIV infected individuals. DESIGN: Cross-sectional study comparing performance of various combinations of screening tests for TB against a gold standard diagnosis of TB based on symptoms, chest radiograph (CXR), sputum microscopy and culture. RESULTS: Of 899 individuals, 44 (4.9%) had TB. The most sensitive symptom combination (59.1%) was any of night sweats, new or worsening cough or reported weight loss; measured weight loss > 5% or abnormal CXR increased sensitivity to 90.9%. Sputum microscopy did not increase sensitivity further, but including World Health Organization HIV clinical staging or CD4 count did. As the specificity of all these combinations was low, many individuals required further investigation to rule out TB. TB prevalence was high (11.7%) among individuals with a CD4 count < 200/mm3. CONCLUSION: CXR greatly increased the sensitivity of screening for TB in this population. Sputum microscopy conferred no additional benefit among asymptomatic patients with a normal CXR. The high prevalence of TB amongst those with a low CD4 count underlines the importance of screening for active TB prior to commencing TB preventive therapy, and before antiretroviral therapy. PMID- 16704035 TI - The use of environmental factors as adjuncts to traditional tuberculosis contact investigation. AB - SETTING: A 25-year-old university student was diagnosed with cavitary pulmonary and laryngeal tuberculosis following symptoms of underlying cough of 6 months' duration. OBJECTIVES: To estimate the hourly risk of infection (HRI) and examine the role of environmental factors, including room size and ventilation, in modulating this risk. METHODS: Contact investigation. RESULTS: Of 1100 contacts identified, 78.3% (n = 896) received a tuberculin skin test (TST), of whom 27.5% had a positive result. Among 634 Canadian-born contacts tested, 22.7% had a positive TST. The independent risk factors for a positive TST among Canadian-born university students were: > 35 h spent with the index case (adjusted OR 6.6, 95% CI 1.0-44.9) and smaller classroom size (aOR 5.0, 95% CI 1.4-10.0). In the first school term, the HRI among Canadian-born student contacts was 0.9%; in the second term, it was 1.6%. CONCLUSION: There are inherent limitations in generalising findings from an outbreak investigation, due to the considerable variation in the infectiousness of cases. Nevertheless, in situations where the index case has a high degree of infectiousness, and there are numerous contacts with low expected prevalence of infection, the HRI can, together with ventilation measurements, be useful in guiding the extent of contact investigation needed. PMID- 16704036 TI - A case-control study of protection against tuberculosis by BCG revaccination in Recife, Brazil. AB - SETTING: Metropolitan region of Recife, Brazil. OBJECTIVE: To estimate the additional protection against tuberculosis (TB) provided by a second dose of bacille Calmette-Guerin (BCG) vaccine. DESIGN: Case-control study. Cases were cases of TB newly diagnosed by the TB control programme, independent of clinical form. Three matched neighbourhood controls were selected using a systematic routine, starting from the case's address. The matching was within the age groups 7-9, 10-14 and 15-19 years. RESULTS: Analysis was conducted among 169 cases and 477 controls. For the efficacy of BCG revaccination against TB overall, matched (crude) vaccine effectiveness (VE) was -3 (95% CI -50-29) and matched (adjusted) VE was 8 (95% CI -77-52). CONCLUSIONS: This study suggests that a second dose of BCG does not offer additional protection. Revaccination should not be offered. As large numbers of subjects are already vaccinated and vaccine appears to offer some protection in older subjects, further studies with larger sample sizes could investigate the potential efficacy of revaccination with BCG in the age group > or = 15 years. PMID- 16704037 TI - Mortality in a large tuberculosis treatment trial: modifiable and non-modifiable risk factors. AB - SETTING: North America. OBJECTIVES: Tuberculosis (TB) patients in North America often have characteristics that may increase overall mortality. Identifying modifiable risk factors would allow for improvements in outcome. DESIGN: We evaluated mortality in a large TB treatment trial conducted in the United States and Canada. Persons with culture-positive pulmonary TB were enrolled after 2 months of treatment, treated for 4 more months under direct observation, and followed for 2 years (total observation: 28 months). Cause of death was determined by death certificate, autopsy, and/or clinical observation. RESULTS: Of 1075 participants, 71 (6.6%) died: 15/71 (21.1%) HIV-infected persons, and 56/1004 (5.6%) non-HIV-infected persons (P < 0.001). Only one death was attributed to TB. Cox multivariate regression analysis identified four independent risk factors for death after controlling for age: malignancy (hazard ratio [HR] 5.28, P < 0.0001), HIV (HR 3.89, P < 0.0001), daily alcohol (HR 2.94, P < 0.0001), and being unemployed (HR 1.99, P = 0.01). The risk of death increased with the number of independent risk factors present (P < 0.0001). Extent of disease and treatment failure/relapse were not associated with an increased risk of death. CONCLUSIONS: Death due to TB was rare. Interventions to treat malignancy, HIV, and alcohol use in TB patients are needed to reduce mortality in this patient population. PMID- 16704038 TI - Tuberculosis trends in Madrid, 1994-2003: impact of immigration and HIV infection. AB - SETTING: Tuberculosis (TB) cases reported from nine districts of Madrid, where the percentage of immigrant population varied from 1.9% in 1996 to 12.2% in 2003. OBJECTIVE: To describe the trends in TB incidence from 1994 to 2003. DESIGN: Observational study. RESULTS: Between 1994-1995 and 2002-2003, the TB rate decreased from 48.5 (95% CI 45.8-51.1) to 23.3 per 100000 population (95% CI 21.5 25.1) (P < 0.001). The percentage of TB cases co-infected with HIV decreased from 55.9% in 1994 to 14.3% in 2003 (P < 0.001), whereas TB cases in foreigners increased from 2.6% in 1994 to 33.7% in 2003 (P < 0.001). CONCLUSION: Although the TB rates showed a marked decrease in the study period, the increasing impact of immigration contributed to slowing down the trend. PMID- 16704039 TI - Drug-resistant tuberculosis in Castilla-Leon, Spain, 1996-2000. AB - SETTING: During 1996-2000, a regional anti-tuberculosis drug resistance survey was conducted in Castilla-Leon, Spain. OBJECTIVE: To determine the incidence of drug-resistant tuberculosis (TB) in newly treated human immunodeficiency virus (HIV) negative and HIV-positive TB patients. DESIGN: Nine hundred and eighty-five Mycobacterium tuberculosis strains isolated from HIV-negative (926) and HIV positive (59) patients were studied (one strain per patient). Univariate and multivariate analyses were used to determine the prevalence of drug resistance in high-risk groups. RESULTS: Thirty-eight isolates (3.8%) showed resistance to one of the following drugs: streptomycin (S), isoniazid (H), rifampicin (R) or ethambutol (E). Of these, 36 (3.9%) were from HIV-negative and 2 (3.4%) from HIV positive patients. The rate of drug resistance among HIV-negative patients was 1.2%, 2.0%, 0.3% and 0.8%, respectively, for S, H, R and E, and for HIV-positive patients it was 3.4%, 0%, 0% and 1.7%. Among the HIV-negative patients, monoresistance was observed in 32 (3.4%) strains and resistance to both H and R (multi-drug resistance) was detected in one. CONCLUSION: The incidence of primary drug resistance in the surveyed area was low and increased resistance was not observed in the HIV-positive group (P = 0.99). Routine surveillance of drug resistance is recommended by the TB control programme in representative patient populations to optimise treatment regimens. PMID- 16704040 TI - Resistance of Mycobacterium tuberculosis isolates in different populations in Greece during 1993-2002. AB - OBJECTIVE: To document the prevalence of resistance of Mycobacterium tuberculosis to isoniazid (INH), rifampicin (RMP) and both combined (multidrug resistance [MDR]) in Greece from 1993 to 2002. DESIGN: We studied a single sputum sample per patient from 4108 patients referred to the Greek National Referral Centre for Mycobacteria. Patients were divided into native Greeks, immigrants and repatriated Greeks originating from the former Soviet Union. Prior treatment status was not recorded. RESULTS: A statistically significant increase in resistance to INH and RMP and MDR was noted comparing the years 1993-1997 to 1998 2002 (P < 0.0001). Resistance to INH and RMP and MDR rose from 5.6% to 7.71%, from 1.57% to 4.49% and from 1.23% to 3.98%, respectively, among native Greeks and from 23.63% to 32.91%, from 6.36% to 15.19% and from 6.36% to 13.92% among repatriated Greeks. Smaller changes were seen among immigrants (from 15.43% to 9.57% for INH, from 5.51% to 6.12% for RMP and from 5.71% to 5.32% for MDR). CONCLUSION: We documented an increase in M. tuberculosis resistance to INH and RMP, and MDR. This was mainly limited to native and repatriated Greeks. Although this is likely the result of immigration and of mismanagement of index cases in Greece, molecular methods are needed to better describe the situation. PMID- 16704041 TI - Alcohol and drug use disorders, HIV status and drug resistance in a sample of Russian TB patients. AB - SETTING: Alcohol use, tuberculosis (TB) drug resistance and human immunodeficiency virus (HIV) risk behavior are of increasing concern in Russian TB patients. DESIGN: A prevalence study of alcohol use and HIV risk behavior was conducted in a sample of 200 adult men and women admitted to TB hospitals in St Petersburg and Ivanovo, Russia. RESULTS: Of the subjects, 72% were men. The mean age was 41. Active TB was diagnosed using a combination of chest X-ray, sputum smears and sputum cultures. Sixty-two per cent met DSM-IV criteria for current alcohol abuse or dependence. Drug use was uncommon, with only two patients reporting recent intravenous heroin use. There was one case of HIV infection. The mean total risk assessment battery score was 3.4. Depression was present in 60% of the sample, with 17% severely depressed. Alcohol abuse/dependence was associated with an eight-fold increase in drug resistance (OR 8.58; 95% CI 2.09 35.32). Patients with relapsing or chronic TB were more likely to meet the criteria for alcohol abuse/dependence (OR 2.56; 95% CI 1.0-6.54). CONCLUSION: Alcohol use disorders are common in patients being treated for active TB, and are associated with significant morbidity. Additional surveys are needed to examine the relationship between alcohol use disorders and anti-tuberculosis drug resistance. PMID- 16704042 TI - Reviving an old idea: can artificial pneumothorax play a role in the modern management of tuberculosis? AB - OBJECTIVE: To determine the usefulness of artificial pneumothorax (AP) in the management of pulmonary tuberculosis (PTB) patients when anti-tuberculosis treatment is ineffective. DESIGN: We evaluated the outcome of therapy in 214 patients with cavitary PTB bacteriologically confirmed by culture treated during 1998-2004, 78.9% of whom had multidrug resistance. AP was applied in 109 patients (56 newly diagnosed TB and 53 retreatment cases). A control group consisted of 105 patients (respectively 55 and 50) treated without AP. The average period of AP application was 4.5 months for newly diagnosed patients and 9 months in retreatment cases. Anti-tuberculosis treatment regimens in both groups were based on drug susceptibility test results. RESULTS: Culture negativity was achieved in patients treated with AP in all new cases and in 81.1% of retreatment cases. Cavity closure occurred in 94.6% and 67.9% respectively. In the control group, culture negativity was achieved in respectively 70.9% and 40.0%, and cavity closure occurred in respectively 56.3% and 24.0%. CONCLUSION: AP considerably improved the treatment outcome in both newly diagnosed and retreatment patients. This procedure can be considered a useful addition in managing certain patients with cavitary TB, particularly those with drug resistance. PMID- 16704043 TI - Recognising tuberculosis in the lower lung field: an age- and sex-matched controlled study. AB - OBJECTIVE: Tuberculosis (TB) can sometimes present with consolidation in the lower lung field. This study was conducted to compare the manifestations of lower lung-field TB (LLFTB) and other pulmonary TB. DESIGN: All new culture-proven TB patients with lower-lung-field consolidation from July 2001 through December 2002 were included. Age- and sex-matched TB patients with upper lung involvement were selected as controls. RESULTS: A total of 79 patients with LLFTB were included. Their mean age was 58.8 years; 46 were male. The clinical, radiographic and laboratory findings were similar in the LLFTB and the control groups, except that the LLFTB patients had less cavitation (P = 0.005). Patients with LLFTB were diagnosed (P = 0.051) and treated (P = 0.001) later than the control patients. The calibres of the trachea and both main bronchi were significantly smaller in the LLFTB group (P < 0.001). More patients with LLFTB developed segmental or lobar atelectasis during follow-up (P = 0.028). CONCLUSIONS: The manifestations of LLFTB are non-specific. The lower-lung involvement, the lower incidence of cavitation and the higher probability of segmental or lobar atelectasis implied that LLFTB was primary TB. A small bronchial calibre probably contributed to its development. PMID- 16704044 TI - Poor attendance at a child TB contact clinic in Malawi. AB - SETTING: Child tuberculosis (TB) contact clinic, Queen Elizabeth Central Hospital, Blantyre, Malawi. DESIGN: Patients registered with smear-positive pulmonary TB (PTB) were encouraged to bring childhood household contacts to the clinic for assessment and management. Data of TB cases registered over the same period were collected from the Blantyre District TB Office. RESULTS: Attendance at the contact clinic was very poor, representing only 7.7% of all adults registered with smear-positive PTB over 17 months, and was significantly lower for potential male source cases than females (OR 0.36, 95% CI 0.23-0.55, P < 0.001). DISCUSSION: Improved uptake and implementation of child contact management in Malawi is a challenge. PMID- 16704045 TI - Tuberculosis and granuloma formation in patients receiving anti-TNF therapy. AB - In patients receiving anti-tumor necrosis factor (TNF) therapy, a probable exacerbation of latent tuberculosis (TB) is a major adverse event. The impairment of granuloma differentiation is considered a characteristic feature of TB in these patients. In this report we present three patients with rheumatic disease who developed TB under infliximab treatment. All of them had typical granulomas on the biopsy specimens, indicating that the expected impairment of granuloma formation is not always the case. The notion of granuloma-free TB in patients receiving anti-TNF therapy could shift a clinician's path away from performing a biopsy, thus delaying the establishment of a correct diagnosis. PMID- 16704046 TI - Ecotoxicology of anticholinesterase pesticides: data gaps and research challenges. PMID- 16704047 TI - Acute and chronic effects of pulse exposure of Daphnia magna to dimethoate and pirimicarb. AB - Short-term (<48 h) and long-term (21 d) effects of dimethoate and pirimicarb were studied in Daphnia magna exposed to pulses of 0.5 to 8 h in duration. During a 21 d postexposure observation period, the following parameters were monitored: Mortality, mobility, day for first offspring, animal size, weight of offspring and adults, and number of offspring produced. In general, animals exposed to a single pulse of dimethoate or pirimicarb regained mobility after 24 to 48 h in clean media. Animals exposed to repeated pulses of dimethoate did not recover mobility during a 48-h postexposure observation period, and mortality was significantly increased. Animals exposed to two pulses of pirimicarb showed less recovery of mobility compared with those exposed to one pulse. Exposure of D. magna to 30 mg/L of dimethoate or 100 microg/L of pirimicarb for 2 to 6 h resulted in a significant reduction in the number of offspring and in the average weight of offspring. The average body length was reduced after pulse exposure to 30 mg/L of dimethoate for 3 h or 70 microg/L of pirimicarb for 4 h, and these exposure concentrations caused a delay in the day for first offspring at exposure durations of 2 to 6 h. The most important new findings in the present study are that short-term (<4 h) pulse exposure of neonates to acetylcholinesterase inhibiting pesticides caused reproductive damage in D. magna and that repeated pulse exposure significantly increased mortality in animals that apparently had recovered after a single-pulse exposure. PMID- 16704048 TI - Effects of the organophosphate paraoxon-methyl on survival and reproduction of Daphnia magna: importance of exposure duration and recovery. AB - The aim of this study was to determine the influence of exposure duration (1 h, 24 h, continuous) to paraoxon-methyl on the magnitude of lethal and sublethal effects, the shape of the concentration-response relationships and the recovery processes in Daphnia magna. Survival was more severely reduced in the continuous than in the pulse exposure regimes. The lethal concentrations (3d median lethal concentration [LC50] values) were 233, 2.33, and 1.14 microg/L after 1-h, 24-h, and continuous exposure, respectively. The shapes of the concentration-response relationships for survival were significantly different after 1 h of exposure than after 24-h and continuous exposure. Indeed, the slopes of the curves defined by the ratios LC90/LC10 (ratio of 90 and 10% lethal concentrations) were 100, 1.74, and 1.97 for 1-h, 24-h, and continuous exposure, respectively. The large difference between 1 h and longer durations of exposure shows that the population is partially affected (10-90%) over a much broader range of concentrations when exposure is short. Negative effects on reproductive outputs occurred mostly at concentrations affecting partly the survival and therefore also over a broad range of concentrations after 1 h of exposure. However, these effects were only transient in the pulse exposure regimes as individual performances recovered. By contrast, reproductive outputs of survivors exposed continuously remained impaired. These results suggest that a refined risk assessment should consider exposure duration because it influences the magnitude of effects and recovery. PMID- 16704050 TI - Influence of body size on swimming performance of four species of neonatal natricine snakes acutely exposed to a cholinesterase-inhibiting pesticide. AB - Locomotor performance is an important fitness-related trait in reptiles because of its potential influence on prey capture and predator avoidance. Because cholinesterase-inhibiting pesticides disrupt neuromuscular signaling, reduction in performance seems to be a logical translation of this biochemical disruption to the organism level. In the present study, we compared the swimming performance of four species of natricine snakes acutely exposed to a formulation of carbaryl to determine whether neonatal body size or skin permeability influences responsiveness. Exposure to high concentrations of carbaryl (2.5-5.0 mg/L) resulted in reduced swimming performance in all four species of snakes, and species responded similarly to the pesticide once body size was accounted for allometrically. Using traditional methods in physiological ecology to estimate skin permeability (a parameter that influences the dose of contaminant absorbed), we found that water flux across the integument also scaled allometrically with body surface area and, therefore, was similar among species after controlling for this relationship. We suggest that future studies examining the effects of repeated low-dose exposure to cholinesterase inhibitors on performance parameters will be useful in assessing the ecological significance of our findings. PMID- 16704049 TI - Dose-additive inhibition of chinook salmon acetylcholinesterase activity by mixtures of organophosphate and carbamate insecticides. AB - Organophosphate and carbamate insecticides are widely detected in surface waters of the western United States. These chemicals interfere with acetylcholine mediated synaptic transmission in the nervous systems of fish and other aquatic animals via the inhibition of AChE (acetylcholinesterase) enzyme activity. Anticholinesterase insecticides commonly co-occur in the environment. This raises the possibility of antagonistic, additive, or synergistic neurotoxicity in exposed fish, including threatened and endangered species of Pacific salmon. We extracted AChE from the olfactory nervous system of chinook salmon (Oncorhynchus tshawytscha) and investigated the inhibitory effects of organophosphates (the oxon derivatives of diazinon, chlorpyrifos, and malathion) and carbamates (carbaryl and carbofuran), alone and in two-way combinations. We found that the joint toxicity of anticholinesterase mixtures can be accurately predicted from the inhibitory potencies of individual chemicals within a mixture. This indicates that organophosphate and carbamate insecticides are noninteractive in terms of AChE inhibition and that it might be possible to estimate the cumulative neurotoxicity of mixtures by simple dose addition. Because organophosphates and carbamates are likely to have additive effects on the neurobehavior of salmon under natural exposure conditions, ecological risk assessments that focus on individual anticholinesterases might underestimate the actual risk to salmon in watersheds in which mixtures of these chemicals occur. PMID- 16704051 TI - Lethal risk to birds from insecticide use in the United States--a spatial and temporal analysis. AB - We used pesticide use data and previously published models to estimate the lethal risk to birds from insecticides used in U.S. agriculture. Data from the U.S. Department of Agriculture's National Agricultural Statistical Service (NASS, Washington, D.C.) were used to assess how the lethal risk to birds has changed over the period 1991 to 2003 and to compare risk among crop types according to the most recently available surveys. Because the NASS data coverage is incomplete, both with respect to crop and state, we also used a database assembled by the National Center for Food and Agriculture Policy (NCFAP, Washington, D.C.) for the 1997 reference year, to which we added state-specific average application rates for crop/insecticide combinations. For each state/crop/insecticide combination (>6000 entries), we assessed the proportion of crop area on which avian mortality was deemed probable, as well as the extended number of hectares this represented. The crops responsible for most potential bird mortality in the United States were corn and cotton, followed more distantly by alfalfa, wheat, potato, peanut, sugar beet, sorghum, tobacco, and citrus. Other crops represented a higher risk to birds on a per hectare basis. The southeast United States generally had the highest proportion of farmland with a lethal risk to birds. On a positive note, the lethal risk to birds has generally declined over the last decade in most crops, although there are exceptions such as small fruit crops. The reasons for this improvement vary from crop to crop, but usually entail the replacement of older more hazardous products with newer ones with lower acute toxicity to birds. PMID- 16704052 TI - Comparison of the applicability domain of a quantitative structure-activity relationship for estrogenicity with a large chemical inventory. AB - The aim of the present study was to illustrate that it is possible and relatively straightforward to compare the domain of applicability of a quantitative structure-activity relationship (QSAR) model in terms of its physicochemical descriptors with a large inventory of chemicals. A training set of 105 chemicals with data for relative estrogenic gene activation, obtained in a recombinant yeast assay, was used to develop the QSAR. A binary classification model for predicting active versus inactive chemicals was developed using classification tree analysis and two descriptors with a clear physicochemical meaning (octanol water partition coefficient, or log Kow, and the number of hydrogen bond donors, or n(Hdon)). The model demonstrated a high overall accuracy (90.5%), with a sensitivity of 95.9% and a specificity of 78.1%. The robustness of the model was evaluated using the leave-many-out cross-validation technique, whereas the predictivity was assessed using an artificial external test set composed of 12 compounds. The domain of the QSAR training set was compared with the chemical space covered by the European Inventory of Existing Commercial Chemical Substances (EINECS), as incorporated in the CDB-EC software, in the log Kow / n(Hdon) plane. The results showed that the training set and, therefore, the applicability domain of the QSAR model covers a small part of the physicochemical domain of the inventory, even though a simple method for defining the applicability domain (ranges in the descriptor space) was used. However, a large number of compounds are located within the narrow descriptor window. PMID- 16704053 TI - Toxic effects of uranium on Desulfovibrio desulfuricans G20. AB - The toxic effects of U(VI) were studied using Desulfovibrio desulfuricans G20 in a medium containing bicarbonate or 1,4-piperazinediethane sulfonic acid disodium salt monohydrate (PIPES) buffer (each at 30 mM and pH 7). Uranium(VI) toxicity was dependent on the medium buffer and was observed in terms of longer lag times and, in some cases, no measurable growth. The minimum inhibiting concentration was 140 microM U(VI) in PIPES-buffered medium. This is 36-fold lower than that reported previously for D. desulfuricans. For all cases in which D. desulfuricans G20 grew in the presence of U(VI), the final cell protein yield was equivalent to that of the U(VI)-free control. In 24 h, D. desulfuricans G20 (total cell protein, 40 mg/L) removed 50 FiM U(VI) from solution in PIPES buffer, as compared to 96 microM U(VI) in bicarbonate buffer under anaerobic, nongrowth conditions. Even though the solubility of U(VI) was significantly lower in PIPES buffer than in bicarbonate buffer, U(VI) was much more toxic in PIPES buffer than in bicarbonate buffer. Analysis of thin sections of D. desulfuricans G20 treated with 90 microM U(VI) in medium containing PIPES buffer revealed that only a very small fraction of cells had reduced U precipitates in the periplasmic spaces. In the presence of bicarbonate buffer, however, reduced U was observed not only in the periplasm but also in the cytoplasm. Selected-area electron diffraction patterns and crystallographic analysis of transmission-electron microscopic lattice fringe images confirmed the structure of precipitated U in the cell periplasm and cytoplasm as being that of uraninite. These results suggest that U(VI) toxicity and the detoxification mechanisms of D. desulfuricans G20 depend greatly on the chemical forms of U(VI) that are present. PMID- 16704054 TI - Two complementary sides of bioavailability: accessibility and chemical activity of organic contaminants in sediments and soils. AB - Research during the last decade has led to several competing concepts of bioavailability and to many more methods to measure bioavailability. One reason for disagreement is the confusion of two fundamentally different parameters, accessible quantity and chemical activity. The accessible quantity describes a mass of contaminants, which can become available to, for example, biodegradation and biouptake. It can be determined with mild extraction schemes or depletive sampling techniques. The chemical activity, on the other hand, quantifies the potential for spontaneous physicochemical processes, such as diffusion, sorption, and partitioning. For instance, the chemical activity of a sediment contaminant determines its equilibrium partitioning concentration in sediment-dwelling organisms, and differences in chemical activity determine the direction and extent of diffusion between environmental compartments. Chemical activity can be measured with equilibrium sampling devices and, theoretically, is closely linked to fugacity and freely dissolved concentration. The distinction between accessibility and chemical activity is outlined, and the benefits and limitation of both endpoints are provided. Finally, examples of how to measure and apply them are presented. PMID- 16704055 TI - Organohalogen contaminants and metabolites in beluga whale (Delphinapterus leucas) liver from two Canadian populations. AB - Contaminants described as organochlorines (OCs; e.g., polychlorinated biphenyls [PCBs]) are present in tissues of marine mammals, including beluga whales (Delphinapterus leucas), but the complexity of contaminant exposure often is not fully known. The PCBs, OC pesticides, polybrominated diphenyl ether (PBDE) flame retardants, methylsulfonyl (MeSO2)- and hydroxy (OH)-PCB metabolites, and OH PBDEs and methoxylated (MeO)-PBDEs were determined in the liver of beluga whales from two Canadian populations: the St. Lawrence Estuary (SLB; n=6), and western Hudson Bay in the Canadian Arctic (CAB; n=11). The sigmaPCB, sigmaDDT, and sigmaPBDE concentrations were higher (p < 0.05) in SLB versus CAB. Of 18 detectable OH-PCBs in SLB (mainly 4-OH-CB107, 4-OH-CB112, and 4'-OH-CB120), only 4'-OH-CB120 was found in CAB. The sigmaOH-PCB concentrations were less than 0.2% of the sigmaPCBs in both populations but were higher (p < 0.05) in SLB (65 +/- 22 ng/g lipid wt) than in CAB (3.1 +/- 0.5 ng/g lipid wt). The sigmaMeSO2-PCB concentrations were higher in SLB (3801 +/- 1322 ng/g lipid wt) relative to CAB (77 +/- 23 ng/g lipid wt) and were 11 and 4%, respectively, of the sigmaPCB concentrations. Of the 15 OH-PBDEs, only two congeners were detectable, but not quantifiable (notably 2'-OH-BDE 68 and 6-OH-BDE 47), in animals from both populations. Of the 15 MeO-PBDEs, 4'-MeO-BDE 17 and 6-MeO-BDE 47 in the SLB (n=2) and 2'-MeO-BDE 68 and 6-MeO-BDE 47 in the CAB (n=2) had concentrations from 20 to 100 ng/g lipid weight. The OH-PBDEs and MeO-PBDEs most likely are of natural origin and accumulated in beluga whales, whereas the OH-PCBs and MeSO2-PCBs are metabolites derived from accumulated PCBs. Canadian beluga whale liver contains previously unidentified organohalogen contaminants and metabolites and, thus, a complexity of contaminant exposure that may be impacting the health of Canadian beluga whale populations. PMID- 16704056 TI - How quality and quantity of organic matter affect polycyclic aromatic hydrocarbon desorption from Norwegian harbor sediments. AB - The desorption behavior of phenanthrene, pyrene, and benzo[a]pyrene was investigated for three Norwegian harbor sediments and their respective particle size fractions using the Tenax desorption method. Rate constants for rapidly, slowly, and very slowly desorbing fractions were on the order of 10(-1), 10(-2) to 10(-4), and 10(-4) to 10(-6)/h, respectively. Relatively small amounts were present in the rapidly desorbing fractions (F(rapid): < 6% for phenanthrene, 3 19% for pyrene, and 1-12% for benzo[a]pyrene). With the exception of benzo[a]pyrene, these F(rapid) values were generally lower than median F(rapid) values obtained from more than 100 literature values for native polycyclic aromatic hydrocarbons (PAHs) (22% for phenanthrene, 29% for pyrene, and 8% for benzo[a]pyrene). To understand which parameters influence PAH desorption, relations between desorption behavior and the sediment characteristics were investigated. A significant positive correlation was found between the extent of slow and very slow desorption and the ratios of black carbon to total organic carbon, as well as the temperature at which 50 and 90%, respectively, of the organic matter was oxidized, as obtained from oxidation-only Rock Eval analysis. Thus, black carbon-bound PAHs probably desorb slowly and very slowly. Furthermore, significant positive correlations between desorption behavior and the average particle size were observed, which could be explained by retarded intraparticle diffusion. PMID- 16704057 TI - A comparison of alcohol ethoxylate environmental monitoring data using different analytical procedures. AB - Several analytical methods have been developed for analyzing alcohol ethoxylates (AE) in aqueous environmental samples. These methods differ widely in their selectivity and sensitivity for measuring the AE components; that is, they vary in their resolution of alkyl chain length homologs and ethoxymer distributions (degree of ethoxylation for each homolog). Given these differences, AE monitoring results from different studies often are inconsistent and, sometimes, are deemed to be incomparable. To address these differences, three currently available methods for determining AE concentrations in environmental matrices were compared among a common set of wastewater treatment plant samples. These methods included the detection of hydrogen bromide-derivatized homologs by gas chromatography/mass spectrometry, the detection of aqueous homologs by high-pressure liquid chromatography/electrospray mass spectrometry, and the detection of pyridinium derivatized homologs by high-pressure liquid chromatography/electrospray mass spectrometry. Results from the present study showed that all three methods responded differently in determining the complex suite of chemical species that comprise AE in the environment. The collective information, however, allowed a consistent comparison among the methods. This comparison was then used to reevaluate results from previous AE monitoring studies. Results from this reevaluation provided a more realistic profile of both historical AE removal during wastewater treatment as well as the occurrence of AE in U.S. surface waters. PMID- 16704058 TI - Chemical characterization of brominated flame retardants and identification of structurally representative compounds. AB - Three training sets were selected, each consisting of 10 structurally diverse compounds representative of brominated flame retardants (BFRs) that are either in use or have been used. Just three compounds account for nearly all the total production volume of BFRs. In the present study, however, the physicochemical characteristics of a far more structurally diverse set of 65 BFRs was explored using 15 molecular descriptors (including log P, constitutional counts, and semiempirical quantum mechanical parameters) and principal component analysis (PCA). The PCA generated an overview of the structural variation among BFRs, and certain compounds with unique physicochemical properties and specific clusters of compounds with distinct properties were identified. The training-set compounds were selected by applying the condensed information obtained from the PCA and statistical experimental design. The three training sets, which were designated as optimal, practical, and alternative, were selected either to maximize the structural variation (optimal) or to combine structural variation with practical advantages, such as ease of experimental handling and commercial availability (practical and alternative). Inclusion of the suggested compounds in assessments of the persistence, bioaccumulation, and toxicity properties of BFRs and related programs should help to increase our understanding of the effects and environmental fate of these compounds. PMID- 16704059 TI - Anaerobic transformation of a technical brominated diphenyl ether mixture by super-reduced vitamin B12 and dicyanocobinamide. AB - The anaerobic transformation of the technical octabromo diphenyl ether mixture, DE-79, was investigated by incubation with two super-reduced corrinoids, dicyanocobinamide and cyanocobalamin (vitamin B12). The transformation produced 33 brominated diphenyl ethers (BDEs), 23 of which could be assigned to known structures. Within 1 d, the hepta- to nona-BDEs in DE-79 were almost quantitatively transformed with dicyanocobinamide to lower-brominated homologs. Along with the decrease of high-brominated congeners, concentrations of some tetra- to hexabromo congeners initially increased, followed by a decreasing or constant concentration during longer incubations. Super-reduced cyanocobalamin also was able to transform BDE congeners of DE-79. Whereas the transformation rates of hepta- to nona-BDEs of DE-79 by both corrinoids were comparable, tri- to hexa-BDEs were susceptible to further transformation by super-reduced cyanocobalamin. Incubation of DE-79 with either corrinoid produced significant amounts of BDE 49. Because this congener is found in small quantities in both penta- and octabromo technical BDE mixtures, it may be a suitable indicator for reductive debromination of higher-brominated BDEs. Isolated BDE 196 was incubated as well with super-reduced dicyanocobalamine. After 1 d, 13 BDEs could be detected, with BDE 153, BDE 100, and BDE 99 dominating. After 7 d, only tetra BDEs or lower-brominated BDEs were detectable. It could be shown that reductive debromination by super-reduced corrinoids is a possible source for lower brominated BDEs (i.e., BDE 47, BDE 99, BDE 153, and BDE 154). The patterns observed with our bench-scale study design demonstrate that anaerobic transformation in the presence of reducing agents can help to explain the occurrence of environmentally relevant BDE congeners (e.g., BDE 47, BDE 99, and BDE 153). PMID- 16704061 TI - Effects of exposure duration and recovery time during pulsed exposures. AB - In pulsed toxicant exposures, the concentration, duration, and frequency of pulses can change through time. However, the conventional median lethal concentration (LC50) method cannot adequately predict the effects of pulsed exposure, because it is associated with fixed exposure duration and constant concentration and does not include postexposure (latent) mortality. Many studies that tried to address the effects of pulsed exposure only provided qualitative or semiquantitative predictions. Survival time experiments conducted here quantified the effect of exposure duration on latent mortality, and the effect of recovery time between two pulses on mortality during a second pulse also was examined. This was done by exposing amphipods (Hyalella azteca) to two contrasting toxicants, copper sulfate (CuSO4) and sodium pentachlorophenol (NaPCP). In the exposure duration experiments, the amphipods were exposed to two toxicant concentrations for three durations. No significant effect of duration on latent mortality was detected within the experimental range; however, duration still may need to be considered under other conditions. In the recovery time experiments, the amphipods were provided four recovery times, and their survivals during the second exposure were modeled. Recovery time had a significant effect on the second-exposure mortality. Given enough time, the amphipods could recover to a state similar to their original toxicant resistance state. The complete recovery time for CuSO4 was fivefold longer than that for NaPCP. It is important to quantify the effects of latent mortality, pulse duration and concentration, and recovery time for pulsed exposures. Survival analysis provides a better way to address these issues than does the conventional LC50 method. PMID- 16704060 TI - Cytotoxicity and aryl hydrocarbon receptor-mediated activity of n-heterocyclic polycyclic aromatic hydrocarbons: structure-activity relationships. AB - Toxic effects of many persistent organic pollutants (e.g., polychlorinated biphenyls or polychlorinated dibenzo-p-dioxins and furans) are mediated via the aryl hydrocarbon receptor (AhR). Although polycyclic aromatic hydrocarbons (PAHs) and their derivatives also activate AhR, their toxic effects remain to be fully elucidated. In the present study, we used the in vitro H4IIE-luc transactivation cell assay to investigate cytotoxicity and potencies to activate AhR by 29 individual PAHs and their N-heterocyclic derivatives (aza-PAHs). The aza-PAHs were found to be significantly more cytotoxic and more potent inducers of AhR than their unsubstituted analogues. Several aza-PAHs, such as dibenz[a,h]acridine or dibenz[a,i]acridine, activated AhR within picomolar concentrations, comparable to the effects of reference 2,3,7,8-tetrachlorodibenzo-p-dioxin. Ellipsoidal volume, molar refractivity, and molecular size were the most important descriptors derived from the modeling of quantitative structure-activity relationships for potencies to activate AhR. Comparable relative toxic potencies (induction equivalency factors) for individual aza-PAHs are derived, and their use for evaluation of complex contaminated samples is discussed. PMID- 16704062 TI - Effects of sediment-bound zinc contamination on early life stages of the mummichog (Fundulus heteroclitus L.) in the Christina watershed, Delaware, U.S.A. AB - During the last century, the Christina River, the major estuarine river system in New Castle County (DE, USA), has received loadings of organic and inorganic chemicals, primarily from manufacturing facilities. Among the most abundant chemicals is zinc, which has accumulated in sediments at concentrations as high as 5,440 mg/kg. We studied the possible effects of zinc on early life stages of the mummichog (Fundulus heteroclitus), a resident species in the river and watershed. We conducted three different types of exposures. The first was a 96-h median lethal concentration (LC50) test with larvae exposed to waterborne zinc. The second was a larval exposure with zinc-spiked sediments (obtained from the relatively uncontaminated Magothy River in Anne Arundel County, MD, USA). The third was an embryo-larval exposure with Christina River sediments having a gradient of zinc concentrations. The average 96-h LC50 with newly hatched yolk sac larvae was 970 lig/L. In the larval tests, the average 7- and 21-d LC50s were 1154 and 1012 mg/kg, respectively. In the embryo-larval test, no significant difference was found in survival at concentrations between 38.8 and 1098 mg/kg. However, significant reductions were observed in condition factor at concentrations of 582, 799, and 1098 mg/kg. We calculated an average no-observed effects concentration of 579 mg/kg and an average lowest-observed-effects concentration of 849 mg/kg for larval survival. Based on these results, we suggest that zinc in the Christina River may be affecting early life stages of the mummichog. PMID- 16704064 TI - Alternative approaches can greatly reduce the number of fish used for acute toxicity testing. AB - Acute toxicity tests with algae, daphnids, and fish are required for the classification and environmental risk assessment of chemicals. The degree of risk is determined by the lowest of these acute toxicity values. Many ecotoxicological programs are seeking to reduce the numbers of fish used in acute toxicity testing. The acute threshold test is a recently proposed strategy that uses, on average, only 10 (instead of 54) fish per chemical. We examined the consequences of reducing the number of fish used in toxicity testing on the ultimate outcome of risk assessments. We evaluated toxicity data sets for 507 compounds, including agrochemicals, industrial chemicals, and pharmaceuticals from our internal database. Theoretical applications of the acute threshold test gave similar results to those obtained with the standard fish median lethal concentration (LC50) test but required only 12% as many fish (3195 instead of 27,324 fish used for all compounds in the database). In 188 (90%) of the 208 cases for which a complete data set was available, the median effect concentration for algae or daphnids was lower than the LC50 for fish. These results show that replacement of the standard fish LC50 test by the acute threshold test would greatly reduce the number of fish needed for acute ecotoxicity testing without any loss of reliability. PMID- 16704063 TI - Alteration of dissolved cadmium and zinc uptake kinetics by metal pre-exposure in the Black Sea bream (Acanthopagrus schlegeli). AB - We first examined the uptake kinetics of Cd and Zn in the juvenile marine black sea bream (Acanthopagrus schlegeli) over a wide range of ambient Cd and Zn concentrations, and the relationships with metal accumulation (uptake rate and amount of nonexchangeable surface binding) were established for different fish tissues. Both Cd and Zn accumulation in the body increased linearly with exposure time after the initial metal surface binding. The dissolved Cd and Zn uptake rate constants were 2.64 and 6.50 L/kg/d, respectively, and the kinetics followed a first-order process. No evidence of biphasic transport was found, in contrast to the situation in freshwater fish. Viscera were the most important sites of metal uptake, and gills were the second most important sites. The black sea breams were then acclimated at different Cd or Zn concentrations from either waterborne or dietary source for one week, and the alteration of metal uptake kinetics or subcellular distribution and metallothionein (MT) induction were further quantified. The Cd body burden was enhanced up to 8.6- and 49-fold after waterborne and dietary Cd pre-exposure, respectively. Cadmium pre-exposure also altered the tissue-specific subcellular Cd distribution and significantly elevated tissue MT levels. In contrast, the black sea breams were able to regulate Zn accumulation, and waterborne or dietary Zn pre-exposure had only weak influences on Zn body burden and redistribution. Both Cd and Zn pre-exposures enhanced the metal uptake rate constants, whereas the nonexchangeable surface bindings were less impacted by these pre-exposures. We demonstrated a positive relationship between the Cd uptake rate and Cd or MT concentration in the fish. Pre-exposure to metals may substantially modify the kinetics of metal uptake. PMID- 16704065 TI - Long-term signal of population disturbance after pulse exposure to an insecticide: rapid recovery of abundance, persistent alteration of structure. AB - Little is known about the effect of pulse exposure to toxicants on populations when density regulation is present. Yet, for a more realistic risk assessment, it is necessary to include effect and recovery at the population level. Here, we investigate the long-term and delayed effects as well as the subsequent recovery of populations of Daphnia magna. A 24-h pulse of the pyrethroid fenvalerate reduced the abundance at a concentration of 1.0 microg/L and higher. However, abundance recovered and reached control levels after one to two generation times (GTs) following reproduction of surviving individuals (GT = 8 d, from birth until first reproduction). At high concentrations above the acute median lethal concentration (3.2 micorg/L), abundance initially decreased even more strongly but was then elevated compared to control values for an extended period of time. Population structure (size distribution) was affected at lower concentrations than abundance (> 0.8 microg/L). In addition, the alteration of population structure lasted for a long time, so that control levels were approached only after approximately six or seven GTs. Our results show that pulse exposure to toxicants may lead to a long-term alteration of population structure even at sublethal concentrations. Possible mechanisms that sustain the effects of toxicants may be delayed life-history effects on the individual level and elevated competition because of altered population structure on the population level. PMID- 16704066 TI - Contaminated sediment testing with the bivalve Mulinia lateralis: culture refinement for organism availability. AB - Availability of test species for estuarine benthic assessment is limited; therefore, a method was developed by the U.S. Environmental Protection Agency for using the dwarf surf clam (Mulinia lateralis) to identify adverse biological effects of bulk estuarine sediments. A multilaboratory evaluation of the draft method resulted in favorable responses from the participants with respect to general testing and handling of the clam. However, availability of good-quality test organisms was limited. An investigation of culture techniques determined that culture vessel topography dramatically influenced survival, because larvae accumulated in bottom depressions. Conditioning of brood stock was enhanced by algae naturally containing both 20:5n-3 and 22:6n-3 fatty acids. Survival of larvae to metamorphosis at 14 d postspawn was greatly increased by maintenance at a lighting of 734 lux with addition of gradually increasing amounts of Isochyris galbana and Chaetoceros calcitrans. Greater than 50% survival of 600 juveniles to testing size by 14 d postmetamorphosis was accomplished by intermittent delivery of algae 12 times per day in a total of 13 L of seawater. By 21 d postmetamorphosis, an additional 27% achieved testable size. As a result of identification of these culture parameters, test bivalves can be readily available to improve predictions of adverse biological effects on benthic communities beyond those presently determined through amphipod exposures. PMID- 16704067 TI - Short-term responses to cadmium exposure in the estuarine polychaete Laeonereis acuta (polychaeta, Nereididae): subcellular distribution and oxidative stress generation. AB - Some effects of cadmium exposure (100 microg/L for 4, 8, 12, and 24 h) on the estuarine polychaete Laeonereis acuta (Nereididae) were evaluated. This polychaete was able to accumulate cadmium in the body, with the metal stored mainly in the cytosolic fraction (>10 kDa). Activity of the antioxidant enzymes superoxide dismutase, glutathione S-transferase, and glutathione reductase (GR) as well as the total oxyradical scavenger capacity, the glutamate cysteine ligase catalytic subunit gene expression, and the metallothionein-like proteins content were not affected by cadmium at any exposure time tested. Catalase (CAT) activity, however, was significantly lower (p < 0.05) in worms treated with cadmium compared with that in controls after 8 h of exposure. At the same exposure time, lipid peroxide levels were increased (p < 0.05) in worms exposed to cadmium compared with those in control worms. Interestingly, CAT and GR activities decreased over time (p < 0.05) independent of cadmium treatment, which is a result that could be attributed to starvation. The effects caused by cadmium in the present study were observed only after 8 h of exposure, demonstrating that cadmium can generate oxidative stress. PMID- 16704068 TI - Weathering and toxicity of marine sediments contaminated with oils and polycyclic aromatic hydrocarbons. AB - Many sediments are contaminated with mixtures of oil residues and polycyclic aromatic hydrocarbons (PAHs), but little is known about the toxicity of such mixtures to sediment-dwelling organisms and the change in toxicity on weathering. In the present study, we investigated the effects of a seminatural, two-year weathering period on PAH/oil chemistry and toxicity in a marine sediment that had been spiked with three different oils (a gas oil, a lubricating oil, and a crude oil; all tested at five concentrations). Toxicity of bioavailable, pore water accommodated oil/PAH fractions was quantified using a bacterial (Vibrio fischeri) assay and the in vitro chemical-activated luciferase expression assay (DR-CALUX; using conditions to detect PAHs). Results of chemical analyses pointed to (microbial) degradation of all three oils: Sediment oxygen demand during weathering increased with increasing oil concentration, total oil concentrations decreased to between 17 and 29% of initial levels, and resolved n-alkanes were depleted in weathered oil fractions. Furthermore, a shift in the relative importance of different boiling-point fraction ranges of the oils was observed on weathering. Generally, the lowest fraction range (C10-C16) disappeared, whereas the relative proportion of the highest (C28-C40) fraction range increased considerably. Remarkably, for the gas oil, this fraction shift was dependent on the oil concentration in sediment. Similarly, degradation of PAHs was strongly affected by the sedimentary oil content, indicating that the presence of oil stimulated PAH degradation. This phenomenon applied to both low- and high molecular-weight PAHs, although the first group (3- and 4-ring PAHs) was degraded most. Results from the V. fischeri and DR-CALUX assay showed that in most cases, pore-water toxicity decreased on weathering. Combining the assay responses with chemical data indicated that the observed toxicity probably was not caused by the analyzed PAHs but, rather, by specific oil constituents instead. PMID- 16704069 TI - Effects of diethylstilbestrol and ethinylestradiol on gene transcription of very low-density apolipoprotein II in the liver of Japanese quail, Coturnix japonica. AB - Very low-density apolipoprotein II (apoVLDL) is one of the constituents of yolk in avian eggs. The expression of the apoVLDL gene is highly specific to the liver in mature female birds during the egg-laying period but is stimulated by exogenous estrogens in immature male birds. In the present study, we compared the effects of two estrogenic compounds, diethylstilbestrol and ethinylestradiol, on the expression of apoVLDL mRNA in the liver of Japanese quail (Coturnix japonica). Three-week-old, immature male quail were treated with a single intraperitoneal injection of the estrogenic compounds, and the level of apoVLDL mRNA in the liver was measured by gene-specific real-time polymerase chain reaction. Diethylstilbestrol and ethinylestradiol had a similar effect on the level of mRNA, increasing it in a dose-dependent manner. Next, the levels of apoVLDL mRNA in the liver of male embryos, which were developed in fertile eggs laid by quail injected with the estrogenic compounds during yolk formation, were measured. Maternal exposure to ethinylestradiol caused an increase in the mRNA in embryos, whereas exposure to diethylstilbestrol had no effect. These results point out the importance of the route of administration to the evaluation of the estrogenic effects of endocrine disruptors in oviparous species. PMID- 16704070 TI - Sublethal toxicity of trace metals to larvae of the blacklip abalone, Haliotis rubra. AB - The availability of literature regarding sublethal and chronic toxicity of heavy metals to early life stages of marine species is restricted to a few species of invertebrate mollusks. The early life stage of abalone, an important gastropod both environmentally and commercially, has been involved in limited research investigating the effects of heavy metal toxicity. Fertilized eggs of Haliotis rubra were exposed to a range of dissolved nominal concentrations of cadmium, copper, iron, lead, mercury, and zinc in individual solutions for 48 h. After 48 h of exposure, the test was completed by recording survival success and morphological abnormalities of veliger larvae in each heavy metal treatment. The mean 48-h median effective concentrations affecting normal morphological development of veliger larvae determined in this test shows a decreasing order of toxicity of copper (7 mg/L), mercury (21 mg/L), zinc (35 mg/L), iron (4102 mg/L), cadmium (4515 mg/L), and lead (5111 mg/L). PMID- 16704071 TI - Toxicities of dinitrotoluenes and trinitrobenzene freshly amended or weathered and aged in a sandy loam soil to Enchytraeus crypticus. AB - Scientifically based ecological soil-screening levels are needed to identify concentrations of contaminant energetic materials (EMs) in soil that present an acceptable ecological risk at a wide range of military installations. Insufficient information regarding the toxicity of 2,4-dinitrotoluene (2,4-DNT), 2,6-dinitrotoluene (2,6-DNT), and 1,3,5-trinitrobenzene (TNB) to soil invertebrates necessitated toxicity testing. We adapted the standardized Enchytraeid Reproduction Test (International Standardization Organization 16387:2003) and selected Enchytraeus crypticus for these studies. Tests were conducted in Sassafras sandy loam soil, which supports relatively high bioavailability of nitroaromatic EMs. Weathering and aging procedures for EMs amended to test soil were incorporated into the study design to produce toxicity data that better reflect the soil exposure conditions in the field compared with toxicity in freshly amended soils. This included exposing hydrated, EM-amended soils in open glass containers in the greenhouse to alternating wetting and drying cycles. Definitive tests established that the order of EM toxicity to E. crypticus based on the median effect concentration values for juvenile production in either freshly amended or weathered and aged treatments was (from the greatest to least toxicity) TNB > 2,4-DNT > 2,6-DNT. Toxicity to E. crypticus juvenile production was significantly increased in 2,6-DNT weathered and aged soil treatments compared with toxicity in freshly amended soil, based on 95% confidence intervals. This result shows that future investigations should include a weathering and aging component to generate toxicity data that provide more complete information regarding ecotoxicological effects of energetic contaminants in soil. PMID- 16704072 TI - Effects of pulsed copper exposures on early life-stage Pimephales promelas. AB - Effects of pulsed copper exposures were investigated using Pimephales promelas aged less than 24 h in short-term chronic testing (7 or 14 d) with moderately hard synthetic water. Concentrations tested were between the species mean chronic value (22 microg/L at a hardness of 100 mg/L as CaCO3) and the 7-d continuous exposure EC50 for survival (40 microg/L) to examine exposures that were not acutely toxic and representative of actual wastewater discharge permit exceedences. Factors tested included pulse duration, recovery time between pulses, and pulse frequency. Survival was the main endpoint affected in all treatments (analysis of variance, p < 0.05). Effects on fish biomass, independent of survival effects, were observed in only 2 of 86 treatments examined. Fish survival was negatively affected at average copper concentrations between 7 and 50% of the 7-d continuous exposure EC50. Exposures having a 48- to 96-h recovery time between pulses had less effect on fish survival than did treatments with shorter (12-24 h) or longer (>120 h) recovery times. Results suggest that the criteria averaging periods used in the United States, and the averaging periods typically used in wastewater discharge permit limits for copper, may not protect against effects of certain pulsed exposures. PMID- 16704073 TI - Sublethal effects of lead on northern leopard frog (Rana pipiens) tadpoles. AB - Northern leopard frogs (Rana pipiens) were exposed to environmentally relevant concentrations of lead in water (3, 10, and 100 microg/L as lead nitrate) from the embryonic stage to metamorphosis. Survival, growth, deformity, swimming ability, metamorphosis, and lead tissue concentrations were evaluated. Tadpole growth was significantly slower in the early stages (Gosner stages 25-30) in 100 microg/L treatment. More than 90% of tadpoles in the 100 microg/L treatment developed lateral spinal curvature, whereas almost all the tadpoles in the other groups were morphologically normal. Spinal deformity was associated with abnormal swimming behavior. Maximum swimming speed of tadpoles in the 100 microg/L treatment was significantly lower than that in the other groups. No significant effect of lead exposure was found on percentage metamorphosis, snout-vent length, mortality, and sex ratio of metamorphs. Time to metamorphosis was delayed in 100 microg/L treatment. Lead tissue concentrations in the tadpoles ranged from 0.1 to 224.5 mg/kg dry mass, were positively related to lead concentrations in the water, and fell within the range of tissue concentrations of wild tadpoles found in previous studies. According to our results, the current U.S. Environmental Protection Agency freshwater-quality chronic criterion for lead (2.5 microg/L) is appropriate to protect northern leopard frog larvae. PMID- 16704074 TI - Development of a solvent-free, solid-phase in vitro bioassay using vertebrate cells. AB - Miniaturized bioassays offer many advantages in exploring the toxic potential of chemicals, including small sample volumes and compatibility with high-throughput screening. One problem common to miniaturized systems, however, is the loss of test chemicals because of sorption. The idea of the current study was to use the sorption phenomenon in a positive way. It was found that contaminants sorbed to the growth surface in wells of tissue-culture plates or to the surface of selected sorbent bead materials are available to vertebrate cells growing in direct contact with the contaminant-coated surface. The use of beads provided more flexibility with regard to surface area, materials, and assay format. Biosilon, a bead cell-culture carrier made of polystyrene, was found to be most suitable. It supported cell adherence and allowed the detection of reproducible dose-response curves of an increase in cytochrome CYP1A enzyme activity by sorbed polycyclic aromatic hydrocarbons in the rainbow trout (Oncorhynchus mykiss) liver cell line, RTL-W1. The resulting bead assay provides a miniaturized, solvent-free exposure system. Potential future applications include the coupling to environmental sampling, in which the bead material is used as solid receiving phase before serving as a surface for vertebrate cells to attach and respond. PMID- 16704075 TI - Population growth of Daphnia magna under multiple stress conditions: joint effects of temperature, food, and cadmium. AB - Aquatic organisms in the field often are exposed to combinations of stress factors of various origins. Little is known of the interaction between different types of stressors; hence, the predictability of their joint effects is low. Therefore, the present study analyzed the joint effects of temperature, food, and cadmium on the population growth rate of the water flea Daphnia magna. The results revealed that temperature, food, and cadmium, as well as their interactions, were important factors that influenced life-history parameters and, as a consequence, the population growth rate of D. magna. In general, population growth rate increased at high temperature and food level but decreased when cadmium was present. The positive effect of temperature on population growth rate was smallest at limiting food levels. Negative effects of cadmium on the growth rate were enhanced at elevated temperatures, whereas high food levels protected the daphnids from adverse effects of cadmium. To avoid over- or underestimation regarding the toxicity of substances to field populations, results of standard toxicity tests should be applied in a location-specific way. PMID- 16704077 TI - Sensitivity of brain cholinesterase activity to diazinon (BASUDIN 50EC) and fenobucarb (BASSA 50EC) insecticides in the air-breathing fish Channa striata (Bloch, 1793). AB - With the expansion of agricultural areas within the Mekong River Delta in Vietnam, a concurrent, dramatic increase has occurred in agrochemical usage. To date, little consideration has been given to the negative impacts of this agricultural activity on the aquatic resources of the region. Both acute toxicity and subacute effects on brain cholinesterase (ChE) of two of the most commonly used insecticides, diazinon and fenobucarb, on adult native snakehead (Channa striata) were evaluated in a static, nonrenewable system, the environmental parameters of which, such as dissolved oxygen, water temperature, and pH, fluctuated similarly to field conditions. Four levels of insecticides, from 0.008 to 0.52 mg/L (for diazinon) and from 0.11 to 9.35 mg/L (for fenobucarb), were tested to assess the effects on the brain ChE activity of the snakehead up to 30 and 10 d for diazinon and fenobucarb, respectively. Diazinon was highly toxic to this fish species, with a 96-h median lethal concentration (LC50) of only 0.79 mg/L, and it also caused long-term ChE inhibition, with activity still significantly inhibited by 30% after 30 d for the three highest concentrations. Fenobucarb was less toxic to this species, with a 96-h LC50 of 11.4 mg/L. Fenobucarb caused more rapid ChE inhibition but also rapid recovery. The results of the present study indicate an urgent need to regulate the usage of these pesticides in the Mekong River Delta. PMID- 16704076 TI - Selenite transport and its inhibition in the unicellular green alga Chlamydomonas reinhardtii. AB - The influence of time, ambient concentration, and medium composition on selenite (Se(IV)) uptake by the unicellular green alga Chlamydomonas reinhardtii has been investigated. The aims of the performed experiments were to describe the kinetics of accumulation, to characterize transport capacities, to identify key nutrients influencing absorption, and to establish links between speciation and bioavailability. Our results suggested that the adsorbed fraction was negligible compared to the absorbed one. Over the short time scale considered, the absorption was linear with time, with an estimated conductance of approximately 0.2 nmol/m2/h/nM. Uptake was proportional to ambient levels in a broad range of intermediate concentrations (from nM to microM). However, conductances were higher at low concentrations (< nM) and then decreased with increasing concentrations (> microM). These results suggested that a specific but rapidly saturated transport system was involved at low concentrations, coupled with a nonspecific one that was only saturated at high ambient concentrations (approximately mM). The latter could involve transporters used by anionic macronutrients, which is supported by the fact that increasing sulfate and nitrate concentrations induced significant inhibition of Se(IV) uptake. Finally, Se(IV) speciation changes caused by varying pH did not significantly affect bioavailability. PMID- 16704078 TI - Ecological performance of red drum (Sciaenops ocellatus) larvae exposed to environmental levels of the insecticide malathion. AB - Malathion is a highly soluble organophosphate insecticide that is widely used in agriculture and mosquito eradication campaigns. Fish species, such as red drum (Sciaenops ocellatus), that use seagrass beds as nursery areas could be affected by runoff waters contaminated with malathion. We exposed red drum larvae at the size they reach in estuarine nursery areas to environmentally realistic and sublethal levels of malathion (0, 1, and 10 microg/L). We evaluated the effects of such exposure on ecologically significant behaviors (routine swimming and predator evasion), growth, and resting metabolism. Malathion exposure to relatively low but ecologically realistic concentrations did not affect routine behavior, escape behavior, resting metabolic rate, or growth, indicating that reported environmental levels may be safe for young fishes. However, a recent substantial increase in the use of malathion may elevate surface-water concentrations to levels above those tested in the present study. PMID- 16704079 TI - Mercury exposure and reproduction in fish-eating birds breeding in the Pinchi Lake region, British Columbia, Canada. AB - To determine whether Hg from geologic/mining-related sources at Pinchi Lake (BC, Canada) was causing elevated Hg exposure and/or adversely affecting reproduction in fish-eating birds, breeding bald eagles (Haliaeetus leucocephalus) on Pinchi Lake and four nearby reference lakes were sampled for blood and feather Hg concentrations and monitored for reproductive success during the summers of 2000, 2001, and 2002. Eggs of red-necked grebes (Podiceps grisgena) also were collected and analyzed. Mercury levels in species at various trophic levels from Pinchi Lake averaged approximately twice those in the same species from nearby lakes combined, even in the absence of substantial new inputs of Hg to Pinchi Lake over several decades. In Pinchi Lake, Hg concentrations in blood and feathers of eagles and eggs of grebes were significantly higher than those in corresponding samples from reference lakes. However, the mean Hg concentration (0.25 microg/g wet wt) in grebe eggs from Pinchi Lake was substantially lower than accepted threshold levels for reproductive toxicity in most avian species (0.5-1.0 microg/g wet wt). Mercury concentrations in the blood of adult eagles and their chicks were highly correlated (r = 0.91, p = 0.004). Despite elevated Hg exposure in adult eagles nesting on Pinchi Lake (blood Hg concentration, 4.3-9.4 microg/ml), birds appeared to be in good body condition, did not differ significantly in terms of weight from eagles nesting on reference lakes, and exhibited no evidence of obvious abnormal behavior or lack of coordination. Eagle reproductive success and productivity on Pinchi Lake were not significantly different from those on all reference lakes combined (p = 0.483). PMID- 16704080 TI - Comparing median lethal concentration values using confidence interval overlap or ratio tests. AB - Experimenters in toxicology often compare the concentration-response relationship between two distinct populations using the median lethal concentration (LC50). This comparison is sometimes done by calculating the 95% confidence interval for the LC50 for each population, concluding that no significant difference exists if the two confidence intervals overlap. A more appropriate test compares the ratio of the LC50s to 1 or the log(LC50 ratio) to 0. In this ratio test, we conclude that no difference exists in LC50s if the confidence interval for the ratio of the LC50s contains 1 or the confidence interval for the log(LC50 ratio) contains 0. A Monte Carlo simulation study was conducted to compare the confidence interval overlap test to the ratio test. The confidence interval overlap test performs substantially below the nominal alpha = 0.05 level, closer to p = 0.005; therefore, it has considerably less power for detecting true differences compared to the ratio test. The ratio-based method exhibited better type I error rates and superior power properties in comparison to the confidence interval overlap test. Thus, a ratio-based statistical procedure is preferred to using simple overlap of two independently derived confidence intervals. PMID- 16704081 TI - [The effect of oral carcinoma-associated fibroblasts on the invasion of carcinoma cell line]. AB - OBJECTIVE: To observe the effect of oral carcinoma-associated fibroblasts (CAFs) on the invasion of a lingual carcinoma cell line, and to elucidate the role of CAFs in oral squamous cell carcinoma (OSCC) progression. METHODS: Matrigel was used to remodel the basement memberane, and the interaction model between primary oral CAFs and lingual carcinoma cell line Tca8113 was established by Transwell chamber to observe the effect of CAFs on the invasion of Tca8113. RESULTS: Compared with normal fibroblasts (NFs), CAFs promoted more Tca8113 cells to penetrate Matrigel (P < 0.05). CONCLUSION: Oral CAFs can promote the invasion of lingual carcinoma cell line Tca8113, and play a key role in OSCC progression. PMID- 16704082 TI - [Immunohistochemical study of heme oxygenase-1 expressed in human normal dental pulp cell and tissue]. AB - OBJECTIVE: To study the expression of heme oxygenase-1 (HO-1) in dental pulp tissue and to find out the relationship of distribution and function of HO-1. METHODS: 30 pulp tissues were obtained from clinically extracted human healthy premolars and third molars. The expression of HO-1 in dental pulp was detected by means of SABC immunohistochemical technology. RESULTS: HO-1 immunoreactivity was observed in vascular endothelial cells, odontoblasts and some fibroblasts cells. CONCLUSION: The distribution of HO-1 in normal human dental pulp suggests that HO 1 may play an important role in pulp flow regulation, dentin production and its calcifying; it also may play some roles in dental pulp cells metabolism and differentiation. PMID- 16704084 TI - [Seasonal rhythms of parathyroid hormone-related protein (PTHrP) expression in growing rats after functional mandibular protrusion]. AB - OBJECTIVE: To study parathyroid hormone-related protein (PTHrP) expression during forward mandibular positioning and compare it with the expression during natural growth in different seasons. METHODS: Sixty-four SD rats were randomly divided into experimental and control groups. Each group were randomly divided into four groups according to seasons. Immunohistochemical (IHC) methods were used to test the protein expression of PTHrP. Macroscopic and microscopic approach were applied to analyze the results. RESULTS: PTHrP expressed in mandibular condylar cartilage (MCC), the expression was accelerated and enhanced when the mandible was positioned forward. Furthermore, there was a seasonal rhythm in the protein expression of PTHrP in both experimental and control groups. The protein expression in spring group rose more than other groups. CONCLUSION: The functional appliance therapy can enhance the protein expression of PTHrP. The enhancement has a seasonal rhythm, which indicates that for the functional treatment better results can be achieved in spring. PMID- 16704083 TI - [Integrin 3 mRNA changes after orthodontic teeth movement in periodontitis rats]. AB - OBJECTIVE: To study the integrin beta3 mRNA changes after orthodontic treatment on normal teeth and periodontitis teeth in rats. METHODS: 96 adult SD rats of 10 weeks old were randomly divided into normal tooth move-ment group and periodontitis tooth movement group. The rats in the two groups were sacrificed after 0 d, 12 h, 1 d, 3 d, 5 d and 7 d of tooth movement. The alveolar specimens were prepared. The integrin beta3 mRNA were detected using in situ hybridization in the specimens. The OD index of positively stained osteoclasts for integrin beta3 mRNA after orthodontic tooth movement in the two groups were measured and compared. RESULTS: There were weak positive signals on the cytoplasm of osteoclasts in periodontum in both groups after 12 hours and 3 days force activation. No positive signals were detected in the rest samples. There was no difference in the OD of positive stained osteoclasts between normal and periodontitis groups. Strong expressions were present on cells with one or two nuclei in the alveolar marrow. CONCLUSION: It is suggested that integrin beta3 mRNA is related with osteoclasts maturation and migration in orthodontic tooth movement. PMID- 16704086 TI - [Mechanical force distribution on cleft maxillary finite-element models after alveolar and hard palate bone graft]. AB - OBJECTIVE: To investigate the biomechanical force distribution of three different cleft maxillary finite-element models pre- and post-bone graft with specified load to certain area of the models. METHODS: Developing a cleft palate bony model from a 15-year cleft palate male CT scan DICOM data and generating alveolar bone grafted, alveolopalatal bone-grafted cleft maxillary finite-element model was set up through gluing the graft model. Also the pre-grafted model as compared, vector lip force on the anterior and anteriolateral face of the alveolar ridge was applied, and studied the press distribution properties and localized area. RESULTS: The press principal spread along the alveolar ridge and focused on anterior wall of maxillary prior to graft. But the press tended to be evenly distributed after bone grafted, whether alveolar or/and hard palate bone grafted. The grafted bone could resisted the medially deformation of alveolar crest and decreased the shear press to the nasal base bony structure. The map showed no significant differences along with alveolar or/and hard palate bone graft. CONCLUSION: The postoperative lip pressure plays an important role for the deformation and deviation of alveolar ridge. Alveolar bone graft could even the distribution of the stresses and should be emphasized. But the grafted bony palatal appears superior to but no significant mapping and anti-deformation difference with alveolar bone graft. PMID- 16704085 TI - [Construction of eukaryotic expression vector of short hairpin RNA for transforming growth factor-beta1]. AB - OBJECTIVE: To construct the plasmid containing short hairpin RNA (shRNA) of TGF beta1 expression vector. METHODS: Short chain oligonucleotide was designed according to the TGF-beta1 mRNA sequence provided by Genebank, then DNA segment was gained through annealing after chemosynthesis, and then was cloned to pWH1 vector. The recombinant TGF-beta1 shRNA expression vector was evaluated by using enzyme cutting. At last, the constructed TGF-beta1 expression vector was transfected into salivary gland mucoepidermoid carcinoma (Ms) cells by Lipofectomine TM 2000, and its effect on TGF-beta1 expression was observed by RT PCR and immunohistochemistry. RESULTS: Successful construction was identified by enzyme cutting and the constructed plasmid was called pWH1-TGF-beta1. The shRNA and it inhibited the TGF-beta1 mRNA and protein expression effectively. CONCLUSION: The constructed TGF-beta1 shRNA expression vector can block the TGF beta1 expression in salivary gland mucoepidermoid carcinoma cells. PMID- 16704087 TI - [Functional activity mapping during anticipation of dental pain]. AB - OBJECTIVE: To reveal functional activity mapping during anticipation of dental pain with non-invasive fMRI technique, and to offer therapeutic possibilities for treating chronic oral facial pain psychologically. METHODS: 8 right handed patients with dentinalgia of the right maxillary bicuspid were included in this study. Block design was adopted, BOLD level during anticipation epoch (on) was contrasted with that of rest (off). Functional MRI scan covering the whole brain was carried out. The fMRI data were analyzed by SPM2 software with t-test to generate the activation map. RESULTS: Increased BOLD signals during dental pain anticipation were found dominantly on the left side in inferior, middle frontal gyrus/BA10,46, postcentral gyrus/BA2,3, middle frontal gyrus/BA8, precentral gyrus/BA44 and cerebellum. CONCLUSION: Anticipation of dental pain by itself can activate brain regions, especially prefrontal areas, SI, SMA and cerebellum. The results of pain anticipation are consisted with previous studies except that there is no activation in insula and cingulate cortex. Sensory, motor, cognitive and emotional activation in dental pain anticipation may indicate that brain nociceptive network can be affected by hint. PMID- 16704088 TI - [Clinical application and processing of separate movable partial denture]. AB - OBJECTIVE: This report is to introduce the processing method of the separate movable partial denture and get an initial clinical evaluation. METHODS: The separate movable partial denture was developed as a kind of special design in utilizing double base plate to separately get support for bearing stress and get the stability of retention. RESULTS: The clinical application indicated that the separate movable partial denture has effective function of common movable partial denture as well as has a good function of natural human teeth protection, especially for those patients with I -II tooth loose. CONCLUSION: The separate movable partial denture is the ideal choice of treatment for those patients who are suffering I - II teeth loose. This type of movable partial denture was considered effectively protect natural human teeth and extend the lifetime of natural tooth long-term and further observation in clinical care on this type of denture is needed. PMID- 16704089 TI - [Analysis on occult micrometastasis in levels III - IV of cN0 neck in patients with oral tongue squamous cell carcinoma]. AB - OBJECTIVE: To investigate the frequency of micrometastasis in levels lII - IV of clinical negative neck (cN0) in patients with squamous cell carcinoma (SCC) of oral tongue, and to discuss the management of cervical lymph node for cN0 tongue SCC. METHODS: A total of 471 cervical lymph nodes derived from 25 patients with cN0 tongue SCC, including 263 lymph nodes in level III and 208 lymph nodes in level IV, were included in this study. All lymph nodes were re-examined by anti cytokeratin (CK) immunohistochemical staining combined with semi-serial section per 500 microm. RESULTS: Among the 25 cases, seven patients were confirmed harboring metastasis in 11 lymph nodes of level III, and no positive lymph node in level IV was detected by routine hematoxylin-eosin (HE) staining. 11 positive lymph nodes in level IIl, which confirmed by HE staining, were also detected by immunohistochemical staining with CK combined with semiserial section. Among the 460 cervical lymph nodes in which HE staining did not show metastasis, only one lymph node in level III harboring a 2.0 mm x 1.5 mm micrometastasis was detected by immunohistochemical staining with CK, and no positive lymph node in level IV was detected by immunohistochemical staining with CK. CONCLUSION: The frequency of occult metastasis in level IV was very low, so it seemed unnecessary to dissect level IV for all patients with cN0 tongue SCC. PMID- 16704090 TI - [Antifugal susceptibility testing and antifugal traditional Chinese medicines screening of oral Candida isolated from head and neck cancer patients treated with radiotherapy or chemotherapy]. AB - OBJECTIVE: To evaluate the sensitivity and resistance of pathogenic oral Candida spp. isolated from head and neck cancer patients treated with radiotherapy or chemotherapy to antifungal agents. To screen antifugal agents from Chinese traditional and herbal drugs by NCCLS M27-A2 method. METHODS: Using YBC Test Kit to identify 20 clinical oral Candida isolated from head and neck cancer patients treated with radiotherapy or chemotherapy. The in vitro susceptibilities of 20 oral Candida spp. to 5-flucytosine (5-FC), itraconazole (ITR), fluconazole (FLU), the extracts of 6 Chinese traditional and herbal drugs (caltrop, honeysuckle flower, dandelion, green tea, pine bark, red trefoil) and utility componets of 7 Chinese traditional and herbal drugs (sophorcarpidine, aloperine, archin, glycyrrhizic acid, glycosides of white peony root, glycosides of baikal skullcap root, hydrochloric berberine) were determined by NCCLS M27-A2 method. RESULTS: The proportion of no-C. albicans in all Candida spp. were 25%. All strains were sensitive to 5-flucytosine, 25% stains were resistant to fluconazole and 40% stains were resistant to itraconazole. In all agents from Chinese traditional and herbal drugs, glycosides of white peony root and hydrochloric berberine (C20H18CINO4) exhibited antifungal activity, especially to C. glabrates. CONCLUSION: The proportion of no-C. albicans in all oral Candida spp. isolated from head and neck cancer patients treated with radiotherapy or chemotherapy was pretty high. NCCLS M27-A2 micro-dilution method is a reliable and reproducible method and can be used to screen antifugal agents from Chinese traditional and herbal drugs. PMID- 16704091 TI - [Comparison of straight wire appliance versus edgewise appliance in orthodontic treatment outcome]. AB - OBJECTIVE: To investigate the difference between straight wire appliance and edgewise appliance treatment outcome. METHODS: 80 cases finished with two-arch straight wire or edgewise appliance between 1994 and 2001 were selected. The PAR index on pre- and post-treatment dental casts and PAR score was recorded respectively. According to the same pre-treatment PAR score and extraction pattern, the samples were divided into two groups, straight wire group and edgewise group, and each group had 40 cases. The data were analyzed statistically. RESULTS: Estimated with the post-treatment PAR score of posterior teeth alignment, the treatment with straight wire appliance was lower than that with edgewise appliance (P < 0.01). There was no statistical difference in other recorded PAR score between two groups. For the mean treatment duration, straight wire appliance (3.95 months) was shorter than edgewise appliance. CONCLUSION: As for the posterior teeth alignment, the straight wire appliance was more accurate, and the straight wire appliance was more efficient than edgewise appliance. PMID- 16704092 TI - [Analysis on differences between soft-tissue and hard-tissue profile in malocclusions]. AB - OBJECTIVE: To study the characteristics of soft-tissue integument and the differences between soft-tissue and hard-tissue topography in malocclusions. METHODS: 144 female patients, 12-15 years old, were selected. They were divided into class I, class II and class III groups according to the value of angle ANB which was measured on the pre-treatment cephalographs. Each group had 48 patients. Each patient had same type of skeletal pattern and occlusal pattern, full set of permanent teeth and none of cranofacial soft-tissue and hard-tissue diseases. 4 pairs of measurements describing soft-tissue and hard-tissue sagittal facial pattern and the prominence of lips and incisors were measured on each cephalograph. They were angle SnNsB', angle ANB, angle NsSnPos, angleNAPo, UL SnPos, UI-APo, LL-SnPos and LI-APo. The distribution of soft-tissue sagittal facial pattern in each skeletal group was analyzed. The differences between angle SnNsB' and angle ANB, angle NsSnPos and angle NAPo, UL-SnPos and UI-APo, LL-SnPos and LI-APo were calculated in each patient. Then we calculated the means and the ranges of these differences in each group, the probability of positive and negative difference between each pair of measurements in each group were calculated too. Chi2 test on those probabilities were performed between the three groups. The mean difference between each pair of measurements was then analyzed by ANOVA between the three groups. RESULTS: The disharmony between soft-tissue and hard-tissue sagittal facial pattern was found in 20%-30% of malocclusion patients. There were more or less differences between soft-tissue and hard-tissue topography and the ranges of their variation were quite wide. The soft-tissue integument increasingly tended to augment the convexity of soft-tissue facial profile when skeletal pattern varied from class II to class I to class III, at the same time, tended to increase upper lip prominence and decrease lower lip prominence. CONCLUSION: On the average, soft-tissue integument tends to camouflage the abnormality of hard-tissue topography. But as to individual, the relative independence of soft-tissue integument makes it important to notice the influence of soft tissue on treatment planning and results. PMID- 16704093 TI - [Dimensional differences in craniofacial morphologies of prepubescence Class II patients with deep and shallow mandibular antegonial notching]. AB - OBJECTIVE: To identify certain craniofacial characteristics of untreated prepubescence Class II patients with deep and shallow mandibular notching. METHODS: 18 untreated preadolescent Class II patients with deep mandibular notching and 17 untreated preadolescent Class II patients with shallow mandibular notching were selected. Each subject's pretreatment lateral cephalogram was traced. Seventeen angular and eleven linear measurements were made. The means and standard deviations were calculated for each parameter. The significance of difference between mean values was evaluated by the Independent-Samples T test. Differences were considered statistically significant when the P value was 0.05 or less. RESULTS: There was no difference in ANB between two groups, but deep notch cases had more retrusive maxillary and mandibles with shorter maxillary and corpus. In vertical direction, subjects with a deep mandibular notch had a longer lower facial height and more vertically directed mandibular plane angle than did shallow notch cases, these may be caused by the increasing of alveolar height of maxillary, smaller ramus height and greater gonial angle. CONCLUSION: There were some differences in the craniofacial morphologies between deep and shallow notching groups of preadolescent Class II patients. Compared with shallow notching group, the mandibles with the deep notching rotated downward and backward and have more vertical directed tendency, but there have no obviously difference in horizontal relationship of maxillary and mandible between two groups. PMID- 16704094 TI - [The preventive effectiveness in reducing tooth decay and decalcification of different concentration of fluoride toothpaste for orthodontic patients]. AB - OBJECTIVE: To investigate the preventive effectiveness in reducing tooth decay and decalcification of different concentration of fluoride toothpaste for orthodontic patients. METHODS: 86 patients were divided into the first test group and the second test group. The patients of the first test group brushed tooth with 1.1% sodium fluoride and acidulated phosphate gel. The patients of the second test group brushed tooth with 0.243% sodium fluoride in a silica base. The extent of facial tooth decay and decalcification of the twelve upper and lower teeth from right cuspid to left cuspid was scored blindly and independently by four observers after 12 months of product use. The scores were rated either one (having tooth cavity/decalcification) or zero (no tooth cavity/decalcification). Four observer's readings were averaged per tooth, and then per patient for the two treatment groups. RESULTS: After 12 months of product use, the mean caries score of the first test group was 0.326, and the mean caries score of the second test group was 0.490. There was significant difference between them. CONCLUSION: A gel system containing 1.1% sodium fluoride and acidulated phosphate provides a clinically better efficacy in reducing tooth decay and decalcification than does a toothpaste containing 0.243% sodium fluoride in a silica base under and adjacent to orthodontic brackets used in orthodontic therapy. PMID- 16704095 TI - [Comparison and analysis of human dentin matrix protein 1 promoter activity in three different cells]. AB - OBJECTIVE: To observe and compare the luciferase activities of different length segments of human dentin matrix protein 1 promoter in human dental pulp stem cells (HDPSC), osteoblasts (OC) and Hela cells. METHODS: The differentlength desired DNA segments were obtained from 2 195 bp Dmp1 promoter cloned by PCR method. The amplified promoter segments with different length were cloned into luciferase report gene vector pGL3-Basic, the correct orientation of those inserts was verified by cutting with two different restrict enzymes. The luciferase activity was observed after different pGL3-PDmp1 vectors were transfected transiently into those three different-type cells. RESULTS: 6 Dmp1 promoter segments with different-length were obtained successfully, and luciferase report gene vectors with different promoter segments were successfully constructed after identified by restriction enzymes cutting. They had different luciferase activities when they were transfected transiently into HDPSC, and the region of -505(-)-193 bp and -935(-)-505 bp could be regarded as the specific promoters of Dmp1 promoter for HDPSC and OC respectively, which could include the basic regulatory elements. CONCLUSION: The correct clone of the upstream of human Dmp1 promoter segments with different length had been obtained, and they had strong luciferase activities in HDPSC and OC, but very low in Hela cell. These results will make an important basis for studying mineralized tissue-specific transcriptional regulation mechanisms of Dmp1. PMID- 16704096 TI - [Effects of mesenchymal stem cells transplantation on cranial suture distraction osteogenesis in growing goats]. AB - OBJECTIVE: To observe the effects of mesenchymal stem cells (MSCs)on cranial suture under mechanical strain in growing goats. METHODS: 10 growing goats were used in this study. A customized distractor was used for distraction of the coronal suture at a rate of 0.4 mm/day for 8 days. The experimental group(5 goats) was injected with autologous MSCs into the distracted region, whereas the control group (5 goats) with injection of physiological saline. All animals were killed at 4 weeks after the end of distraction. Scanning electron microscopy and histological analysis were taken to observe the samples. RESULTS: 4 weeks after the end of distraction, the cranial sutures in all animals were separated successfully. The new bone formation at the edge of suture in the experimental group was superior to that in the control group. CONCLUSION: Autologous MSCs transplantation may promote the cranial suture distraction osteogenesis in the growing goats. PMID- 16704097 TI - [Effect on apoptosis of Tca8113 of interference of Bcl-2 and HER-2 genes antisense oligodeoxynucleotides transfection]. AB - OBJECTIVE: To investigate the interference and the mechanisms of Bcl-2 and HER-2 genes antisense oligodeoxynucleotides combined transfection in the human tongue carcinoma Tca8113 cell lines. METHODS: There were 6 groups in our study, normal control group, Bcl-2 sense experimental group, HER-2 sense experimental group, Bcl-2 antisense experimental group, HER-2 antisense experimental group, Bcl-2 and HER-2 genes antisense oligodeoxynucleotides combined transfection experimental group. In the different times after liposome-mediated transfection, the cell apoptosis, Bcl-2 and HER-2 expressing level were observed by RT-PCR and electronic microscope. RESULTS: According to the results of combined transfection experimental group, the apoptosis body and apoptosis cells were observed. The expression of genes were decreased statistically in both Bcl-2 and HER-2, respectively. Bcl-2 and HER-2 combined ASODN was superior to single ASODN in the intervention of tongue carcinoma. CONCLUSION: Bcl-2 and HER-2 ASODN can effectively interfere the expression of HER-2 and Bcl-2 genes. The expression of HER-2 and Bcl-2 can be reduced, and the apoptosis of cells can be enhanced. PMID- 16704098 TI - [Study of induction of tumor specific cytotoxic T lymphocyte by using tumor derived exosome]. AB - OBJECTIVE: To investigate whether the exosomes derived from Tca8113 could induce production of tumor-specific T cells when pulsed onto dendritic cells. METHODS: Tca8113 cell was cultured with RPIM1640, isolated and purified the tumor-derived exosomes from the culture supernatants by ultrafiltration with Millipore centrifual filter devices; frozen and thawed Tca8113 cells to get frozen tumor antigens (FTA). The exosomes and FTA was pulsed onto DC generated from normal human peripheral blood mononuclear cell (PBMC) in vitro. The DC pulsed with FAP or exosomes cocultured with the peripheral blood lymphocytes to transform T cell into specific CTL. To observe the killing and wounding activity of CTL, the CTL and Tca8113 cells were mixed at a ratio of 20 to 1, SPCA-1 cells and 95-D cells was evaluated as control group. RESULTS: The CTL induced by DC pulsed with FAP or exosomes had significant activity killing Tca8113 (P < 0.01); Moreover the CTL induced by DC pulsed with exosomes could also kill SPCA-1 cells (P < 0.05), but the CTL induced by DC pulsed with FTA had not this function. CONCLUSION: Exosomes derived from tumour accumulate in culture supernatants. Exosomes are a natural and new source of tumour-rejection antigens, opening up new avenues for immunotherapy against oral cancers. The exosome-specific CTL could kill another kind of tumor, so tumor-derived exosomes may contain shared tumor-rejection antigens. PMID- 16704099 TI - [Quantitative study of osteoclastic related factors in the process of bone reconstruction]. AB - OBJECTIVE: To detect the expression of osteoclast related factors, tumor necrosis factor-alpha (TNF-alpha), receptor activator of NF-kappaB ligand (RANKL) and tartrate-resistant acid phosphatase (TRAP), in the process of bone remodeling. METHODS: 8-week-old male C57BL/6J mice were employed in this study to detect the expression of osteoclast related factors by real time PCR. RESULTS: TNF-alpha, RANKL and TRAP were up regulated in the process of bone remodeling, they reached the peak on day 2, 5 and 10 individually after injury. CONCLUSION: Osteoclast related factors also participate in bone remodeling, which depends on the delicate balance between bone formation and bone resorption. PMID- 16704100 TI - [Screening and identification of metastasis-related gene expression in tongue carcinoma with cDNA microarray assay]. AB - OBJECTIVE: To identify metastasis-associated genes in tongue carcinoma and to better understand the mechanism underlying tongue carcinoma metastasis. To compared mRNA expression profiles of two tongue carcinoma cell strains with high and low metastatic potentials using microarray technology. METHODS: Tca8113 and Tb cells were used as model systems to study the molecular mechanism of tumor metastasis. Two fluorescent cDNA probes labeled with Cy3 and Cy5 dyes were prepared from the mRNA samples of Tca8113 and Tb cells by reverse transcription method. The two color probes were then mixed and hybridized to the cDNA chip constructed by double dots of 1 152 human genes, and scanned at two wave lengths. Differential expression genes from the above two cell lines were analyzed using computer. Then six of the different expression genes were further validated by RT PCR technique. RESULTS: In the 1 152 clones of known genes and expressed sequence tags that were analyzed, 37 showed significantly different (minimum 2 folds) expression levels in two cell lines. Among the 37 genes, 15 were up regulated (with ratio more than 2) and 22 down regulated (with ratio less than 1/2). The results of RT-PCR analysis were coincident with those of microarray assay. CONCLUSION: Some of these genes are known to be involved in human tumor antigen, immune surveillance, adhesion, cell signaling pathway and growth control. It is suggested that the microarray in combination with a relevant analysis facilitates rapid and simultaneous identification of multiple genes of interests and in this study it provided a profound clue to screen candidate targets for early diagnosis and intervention. PMID- 16704101 TI - [Effect on xenograft of nude mouse by combination therapy of nm23-H1 and protein cisplatin]. AB - OBJECTIVE: To study the effect of protein-cisplatin and nm23-H1 therapy on the tumor of nude mouse. METHODS: The 15 BALB/C female mice were divided into three groups, control group, protein-cisplatin group and protein-cisplatin+nm23-H1 plasmid group. Tca8113 were injected into the mice subcutaneously with the concentration of 3.1 x 10(6) cells/mL. After two weeks, the mixture of lipofectin and nm23-H1 was injected around xenograft of nude mouse. After three days, the protein-cisplatin was injected around xenograft. The weight of mouse, the volume and the weight of xenograft were measured. RESULTS: The weight of mouse was lightest in control group. The volume and weight of the transplanted tumor were lightest in nm23-H1 +protein-cisplatin group. CONCLUSION: The combination therapy of nm23-H1 and protein-cisplatin can effectively inhibites the growth of xenograft in nude mouse. PMID- 16704102 TI - [Effect of implant junction on bone growth by X-ray analysis]. AB - OBJECTIVE: To explore the effects of mandibular implant junction on the growth of immature bone. METHODS: Eight Beagle dogs were randomly divided into three groups: control, unjunction and junction. Twelve implants were produced on the mandible of unjunction experimental group and junction experimental group. At three months after implanting, radiographic examination was performed. RESULTS: Three months after implanting, all implants were integrated with bone. None implants was mobile or got off. Radiographic examination demonstrated that the bone lose difference was no significant in junction and unjunction group. CONCLUSION: Mandibular implant connection wasn't effect on the growth of bone. PMID- 16704103 TI - [A survey of the therapeutic reason of orthodontic patients in Luzhou]. AB - OBJECTIVE: To investigate the therapeutic reason of orthodontic patients in Luzhou. METHODS: A questionnaire investigation was made for the following randomly chosen people: 452 teenage patients from orthodontic department of stomatology hospital affiliated to Luzhou Medical College were brought into treatment group. 600 students from Luzhou No. 2 Middle School were brought into non-treatment group. The questionnaire investigation was conducted to two groups. The results were statistically analysed with SPSS 10.0. RESULTS: (1) The main aim for orthodontic treatment was to align the teeth. (2) 37.39% of the patients accepted orthodontic treatment for parent's demand. (3) 76.1% of the patients expected to align the dentition. (4) Most people in treatment group could accept the time and expenses in the orthodontic treatment, but most people of non treatment group could not accept the expenses in the orthodontic treatment. (5) Most people thought appearance was important. CONCLUSION: Aesthetic is the main reason for orthodontic patients. PMID- 16704104 TI - [Analysis of publication of West China Journal of Stomatology from 2000 to 2005]. AB - OBJECTIVE: To analyze the characteristic of publication of West China Journal of Stomatology (WCJS). METHODS: Making use of literature metrology, a comprehensive analysis of the information publication of West China Journal of Stomatology from 2000 to 2005 was made. RESULTS: The quantity of articles published from 2000 to 2005 in West China Journal of Stomatology was 964, the average fund paper ratio was 0.419, 96.78% of articles had more than one author, and the average paper author cooperating degree was 4.01. The authors scattered in 31 provinces. CONCLUSION: West China Journal of Stomatology is one of the most important information source of stomatology. PMID- 16704105 TI - Icons of dentistry: Dr Leon Eisenbud. AB - Dentistry has a long, often well documented history. Evidence of tooth pullings has been discovered in crude carvings on the walls of caves that are over 10,000 years old. The ancient Egyptians, the Athenians, and the early inhabitants of Rome required oral health care; in addition to tooth extractions, they underwent tumor removal, tamponade for hemorrhage, reduction of jaw fractures with gold wire ligatures, cautery using white hot platinum loops, and an additional variety of remedies and nostrums. Pain relief was offered, with courses of treatment as varied as postural change, alteration of ambient temperature, and vegetable and organic medicines in poultices or via oral and rectal routes. Through the centuries, great surgeons and physicians introduced various methods of treatment: Hippocrates codified ethical standards; Maimonides established pragmatic rules for physicians; LeFort categorized facial fractures; Pasteur clarified the need for sterilization; Semmelweis standardized antiseptic conditions in the operating theater; Morton and Wells discovered safer methods of analgesia; Freud explored the theraupeutic uses of narcotics; Roentgen championed X-ray imaging; Curie pioneered the use of chemotherapy; and Barton and Nightingale were models of empathy and patient care. In more recent times, we have profited from the genius of Watson and Crick (DNA); Fleming (penicillin); Venable and Stuck (Chrome-cobalt -molybdenum alloy); Gershkoff and Goldberg (the subperiosteal implant); Chercheve, Branemark, Linkow, Misch, Tatum, and Niznick (innovative root forms, titanium and its alloys, and sinus floor grafting). The 20th century has brought to us phenomenal imaging, breathtaking intrauterine fetal surgery, wildly promising stem cell research, and astonishing CADCAM techniques. We've had great teachers and clinicians who have introduced us to new forms of therapy and advanced methods, including the role of the hemidesmasomes, the essential elements of bone grafting, the importance of microscopic analysis, and the benefits to patients of physical diagnosis by their dentists. To recognize and celebrate some of my heroes' contributions to the health and well-being of humankind, editorials will occasionally appear on these pages that explore their various contributions. PMID- 16704106 TI - The effect of residual calcium in decalcified freeze-dried bone allograft in a critical-sized defect in the Rattus norvegicus calvarium. AB - Demineralized freeze-dried bone allograft (DFDBA), a widely used graft material in periodontal regenerative procedures, is processed with hydrochloric acid in the attempt to expose proteins located within the bone matrixes that are capable of inducing new bone formation. However, the degree of DFDBA demineralization varies between tissue banks, which may have an effect on clinical regeneration. This study uses the critical-sized defect (CSD) model to evaluate the wound healing response to the residual calcium of donor bone. If the percentage of residual calcium in a graft were demonstrated to significantly enhance wound healing, then periodontal patients may benefit from further standardization of human-allograft processing. Sixty adult, male, Harlan Sprague-Dawley rats (Rattus norvegicus) were randomly and equally divided into 4 test groups (ie, DFDBA at 1%, 2%, and 3% to 6% residual calcium levels and FDBA at 23% residual calcium) and a control group (no allograft). An 8-mm-diameter craniotomy was made in the rat calvarium, and polytetrafluoroethylene membranes with pore sizes of 0.50 microm were placed intracranially and ectocranially. Treatment materials were carefully placed into the CSD with a new sterilized dental amalgam carrier. Tetracycline hydrochloride was injected intraperitoneally for labeling new bone growth, and animals were euthanized 12 weeks postsurgery. As a result, histomorphometric bone fill at 12 weeks showed a statistically significant increase in the 2% DFDBA group as compared to all other groups. The authors conclude that a 2% residual calcium level in human DFDBA appears to significantly (P < or = .05) enhance osseous wound healing in the rat calvarium. PMID- 16704107 TI - The role of interfacial mechanics in the prediction of global mechanical behavior of a bioactive composite: an in vitro study. AB - A bioactive bone-tissue substitute, hydroxyapatite (HA)-polymethylmethacrylate (PMMA) with the addition of a copolymer coupling agent, was examined in vitro to determine the influence of the coupling agent on the local mechanical properties of the system before and after simulated biologic conditions. Nano-indentation of the cross-sectional interface between the HA and PMMA of the composite was studied. The fracture mechanism and position of each indent mark were analyzed at up to 5000x magnification under field-emission, environmental-scanning electron microscopy. The local interfacial results were compared with global quasistatic compression test results. It was found that nano-indentation of the interface could predict changes in global mechanical behavior of the composite. Both interfacial and global Young's moduli were reduced after immersion in the simulated biologic media. Although the coupling agent improved the interfacial and global mechanical properties before and after 24 hours in in vitro immersion, it did not affect the surface bioactivity of the system, as shown in the measurement of calcium and phosphate concentration uptake. Thus, nano-indentation is a sensitive technique for examining interfacial mechanics and mechanical consequences of biologic reactivity of composite materials. PMID- 16704108 TI - Enhancement of osseointegration by generating a dynamic implant surface. AB - Preparations of autologous plasma rich in growth factors (PRGF) are used to promote healing and tissue regeneration. We seek to determine whether covering the titanium implant surface with this preparation could enhance osseointegration. The interaction of PRGF with the surface of titanium implants was examined by environmental scanning electron microscopy (ESEM). A total of 23 implants were placed in the tibiae and radii of 3 goats; 13 implants were inserted after covering the surface and filling the alveolus with PRGF, and 10 more implants were inserted following a conventional protocol and served as controls. Histomorphometric analysis of the bone-implant interface was performed after 8 weeks. Finally, 1391 implants were placed in 295 patients after bioactivating the surface with PRGF. Stability and implant survival were evaluated. The implant surface adsorbed the protein-rich material as shown by ESEM. In the animal study, osseointegration was enhanced when the surface was covered with PRGF as shown by histomorphometry (bone-implant contact: 51.28% +/- 4.7% vs 21.89% +/- 7.36%; P < .01). Finally, studies in patients showed that 99.6% of the implants treated with PRGF were well osseointegrated. Clinical use of this technique in oral implantology can improve the prognosis. PMID- 16704109 TI - Cone beam computerized tomography-based dental imaging for implant planning and surgical guidance, Part 1: Single implant in the mandibular molar region. AB - Computerized tomography (CT)-based dental imaging for implant planning and surgical guidance carries both restorative information for implant positioning, as far as trajectory and distribution, and radiographic information, as far as depth and proximity to critical anatomic landmarks such as the mandibular canal, maxillary sinus, and adjacent teeth. This case report describes a systematic approach to the planning and surgical placement of a single implant-supported crown, utilizing CT-based dental imaging for implant planning and surgical guidance. The simple steps result in the accurate transfer of critical radiographic information to the surgical site. PMID- 16704110 TI - Use of mini implants for replacement and immediate loading of 2 single-tooth restorations: a clinical case report. AB - Numerous long-term studies have shown that treatment with dental implants can provide edentulous patients with a more stable alternative to complete dentures and partially edentulous patients with a more conservative form of tooth replacement than conventional fixed partial dentures. Until recently, commercially available dental implants have been limited to diameters ranging from 3.0 mm to 7.0 mm. Although this range of diameters has been able to address most clinical needs, partially edentulous patients who could not accommodate a 3.0-mm-diameter implant without damaging adjacent dental structures were excluded from implant therapy. This article reports on the surgical treatment and immediate restoration of a patient who received mini implants that were 2.4 mm in diameter. PMID- 16704111 TI - Lateral bone condensing and expansion for placement of endosseous dental implants: a new technique. AB - Placement of endosseous dental implants can be a problem due to bone resorption if the patient has been missing teeth for a considerable period of time. In the literature, bone-grafting techniques have shown variable results. Additionally, bone grafting requires a longer treatment time and a need for a second surgery, and it adds significant cost to the treatment. These factors often discourage patients from having dental-implant treatment. Another technique for placement of dental implants in narrow bone ridges is repositioning and remodeling of alveolar bone by condensing and expansion with the help of bone osteotomes. This article presents 2 cases, 1 in the maxilla and 1 in the mandible, for placement of endosseous dental implants with the use of a new bone-expansion osteotome kit that utilizes a screw-type configuration for bone condensing and expansion. PMID- 16704112 TI - Alterations in DNA methylation and genome structure in two rice mutant lines induced by high pressure. AB - By using high-pressure treatment, two mutant lines were obtained from a genetically stable japonica rice cultivar Bijing38. Genomic DNA of the mutant lines, together with the original line (Bijing38), was either undigested or digested by Hpa IIMsp I, and then subjected to molecular analysis using two markers, ISSR and RAPD. Results indicated that changes in the PCR amplification profiles of both markers are apparent in the two mutant lines compared with the original rice cultivar, suggesting that there had been both sequence changes and DNA methylation modifications in the mutant lines. Southern blot analysis using diverse sequences, including two cellular genes (S2 and S3), a set of retrotransposons (Osr7, Osr36, Tos19 and more), and a MITE transposon family (mPing and Pong), confirmed the results, and indicated that changes in DNA methylation pattern, genomic structure, and possible activation of some transposons indeed occurred in the mutant lines. Moreover, these changes are stably maintained through selfed generations and in different organs. Thus, our results indicate that it is possible to obtain stable mutants in rice by high pressure treatments, and the molecular basis of the mutants may include both genetic and epigenetic changes. Therefore, high hydrostatic pressure seems a promising approach for plant mutagenesis. PMID- 16704113 TI - Isolation of the promoter of a cotton beta-galactosidase gene (GhGal1) and its expression in transgenic tobacco plants. AB - Beta-galactosidases (EC 3.2.1.23) constitute a widespread family of glycosyl hydrolases in plants and are thought to be involved in metabolism of cell wall polysaccharides. A cDNA of the cotton (Gossypium hirsutum) beta-galactosidase gene, designated GhGal1, has previously been identified and its transcripts are highly abundant at the elongation stage of the cotton fiber. To examine the temporal and spatial control of GhGal1 expression, a transcriptional fusion of the GhGal1 promoter region (1770 bp) with the beta-glucuronidase (GUS) reporter gene was introduced into tobacco plants by the Agrobacterium infection method. The resulting transgenic plants showed higher GUS activity of fruit in the transgenic plants than that in the negative and positive controls. Histochemical localization of GUS activity demonstrated that the expression of the GUS gene could be found in the meristem zones of roots, cotyledons, vascular tissues, fruit and trichomes in transgenic tobacco plants. Additionally, sequence analysis of the regulatory region also revealed several conserved motifs among which some were shared with previously reported fruit/seed-specific elements and the others were related with trichome expression. These results indicated the temporal and spatial expression characterization of the GhGal1 promoter in transgenic tobacco plants and provided an important insight into the roles of GhGal1 in cotton fiber development. PMID- 16704114 TI - Cloning and sequencing of nifBHDKENX genes of Paenibacillus massiliensis T7 and its nif promoter analysis. AB - A 324 bp of nifH fragment was PCR amplified from Paenibacillus massiliensis T7 using the universal degenerate primers. The PCR-amplified nifH fragment was labeled with DIG and then used as a probe in Southern blot analysis. Southern blot result showed that there were two positive signals, indicating that there might be two copies of nifH in P. massiliensis T7. A total of 10254 bp DNA sequence containing purD and nifBHDKENX was obtained by five rounds of inverse PCR amplification. The predicted proteins of nifBHDKENX had high homology with those from other nitrogen-fixing bacteria. Only one putative sigma54-dependent promoter sequence was detected upstream of the nifB gene and nifBHDKENX were likely to be organized in one operon. Assays of 3-galactosidase activity of P. massiliensis T7PB carrying a nifB-lacZ fusion under different concentrations of NH4+ and O2 showed that the expression of nifB-lacZ was strongly inhibited by O2. PMID- 16704115 TI - The effects of K+ growth conditions on the accumulation of cesium by the bacterium Thermus sp. TibetanG6. AB - The accumulation of cesium by the bacterium Thermus sp. TibetanG6 was examined under different K+ growth conditions. The effects of external pH and Na+ on the accumulation of cesium were also studied, and the mechanism involved was discussed. K+ regimes played an important role in the accumulation of cesium by the strain TibetanG6. The quantity of cesium accumulated (24 h) was much higher in K+-deficient regime than that in K+-sufficient regime. The pH and Na+ had different effects on the accumulation of cesium in the two K+ regimes. IR spectra analyses indicated that the biosorption is a process of homeostasis with cesium initially accumulated on the cell wall. PMID- 16704116 TI - A novel approach for estimating growth phases and parameters of bacterial population in batch culture. AB - Using mathematical analysis, a new method has been developed for studying the growth kinetics of bacterial populations in batch culture. First, sampling data were smoothed with the spline interpolation method. Second, the instantaneous rates were derived by numerical differential techniques and finally, the derived data were fitted with the Gaussian function to obtain growth parameters. We named this the Spline-Numerical-Gaussian or SNG method. This method yielded more accurate estimates of the growth rates of bacterial populations and new parameters. It was possible to divide the growth curve into four different but continuous phases based on changes in the instantaneous rates. The four phases are the accelerating growth phase, the constant growth phase, the decelerating growth phase and the declining phase. Total DNA content was measured by flow cytometry and varied depending on the growth phase. The SNG system provides a very powerful tool for describing the kinetics of bacterial population growth. The SNG method avoids the unrealistic assumptions generally used in the traditional growth equations. PMID- 16704117 TI - Comparison of the virulence plasmid genomes of two strains of Shigella which lost the ability to bind Congo red. AB - We determined and analyzed the Shigella flexneri serotype 5 (pSF5) and S. dysenteriae serotype 1 (pSD1) virulence plasmid genomes. The total length of pSF5 is 136513 bp, including 165 open reading frames (ORFs). Of these ORFs, 133 were identified and 32 of those had no significant homology to proteins with known functions. The length of pSD1 is 182545 bp, including 224 ORFs, of which we identified 181. The remaining 43 ORFs were not significantly homologous to proteins with known functions. The insertion sequence (IS) elements are 53787 bp in pSF5, and 49616 bp in pSD1, which represents 39.4% and 27.1% of the genome, respectively. There are 22 IS element types in pSF5 and pSD1, among which we report ISEc8 and ISSbo6 for the first time in the Shigella virulence plasmid. Compared to pCP301, there are a large number of deleted genes and gene inversions in both pSF5 and pSD1. The ipa-mxi-spa locus in pSF5 is completely absent, and the genes related to the O-antigen biosynthesis are partially missing. In contrast, the above genes in pSD1 are integral, with the exception of virF. The whole genome analysis of the two plasmids shows that the loss of genes related to gene invasion or regulation also obliterates the ability of pPF5 and pSD1 to bind Congo red (Crb). Whether these genes determine the Crb function requires continued investigation. PMID- 16704118 TI - Sequence analysis for the complete proviral genome of reticuloendotheliosis virus Chinese strain HA9901. AB - The genomic DNA extracted from chicken embryo fibroblast (CEF) infected with a Chinese field isolate HA9901 of reticuloendotheliosis virus (REV) was used as the template to amplify the REV proviral genomic cDNA by PCR with 6 pairs of primers according to published sequences. Six overlapping fragments were amplified, cloned into the TA vector and sequenced, including a fragment which was amplified from the circular proviral cDNA and covering both 5'- and 3'-ends. The complete sequence of the whole genome was established and analyzed with a DNAstar software. Comparisons of the sequence with two other strains demonstrated that the genomes of REV were relatively conservative, the homogenecity for all genes or LTR fragments of the 3 strains was over 92%, no matter whether they were isolated from different species and regions in different years. But, the homology of Chinese strain HA9901 to a fowl pox virus-associated strain from Chickens was higher than that to strain SNV isolated from ducks. PMID- 16704119 TI - Identification of one peptide which inhibited infectivity of avian infectious bronchitis virus in vitro. AB - Purified avian infectious bronchitis virus (IBV) was used to screen a random phage display peptide library. After the fourth panning, 10 positive phages were sequenced and characterized. The phages specifically inhibited IBV infectivity in HeLa cells and blocked IBV haemagglutination. One linear peptide "GSH HRH VHS PFV" from the positive phages with the highest neutralization titer was synthesized and this peptide inhibited IBV infection in HeLa as well. The results may contribute to development of antiviral therapeutics for IBV and studying the determinants for viral and cell interaction. PMID- 16704120 TI - Production of porcine cloned transgenic embryos expressing green fluorescent protein by somatic cell nuclear transfer. AB - In the present study, nuclear transferred embryos (NTEs) were reconstructed by using pig fetal fibroblasts as donors and in vitro matured oocytes as recipients. The effects of G418 selection on donor cells, duration of IVM of prepubertal gilt oocytes and oxygen tension in IVM of oocytes were investigated. The results were as follows: (i) When G418 selected cells expressing GFP were used as donors, the cleavage rate of NTEs decreased drastically in comparison to NTEs derived from donors without antibiotic selection (47.5% vs. 71.6%, p < 0.05). For the blastocyst rate, no significant difference was observed between two groups (10% vs. 10.4%, p > 0.05). (ii) The rate of nuclear maturation of oocytes increased significantly when IVM duration time was extended from 36 to 42 h (83.6% vs. 96.7%, p < 0.05). However, no statistical difference was observed between NTEs derived from oocytes of 36 h IVM group and NTEs from oocytes of 42 h IVM group in the rates of cleavage (59.3% vs. 73.6%, p > 0.05) and blastocyst formation (9.3% vs. 13.2%, p > 0.05); (iii) no significant difference was observed between NTEs reconstructed from oocytes matured under lower oxygen (7% O2) tension and NTEs derived from oocytes matured under higher oxygen tension (20% O2) in cleavage rate (70.6% vs. 67.1%, p > 0.05) and blastocyst rate (11.8% vs. 12.3%, p > 0.05). These results suggest that: (i) G418 selection does not have a significant effect on cleavage rate of NTEs expressing GFP. (ii) Nuclear maturation is greatly improved by prolonging IVM duration from 36 to 42 h, while no significant differences were observed for developmental potential of transgenic embryos. Thus IVM 42 h is the better choice in order to obtain maximum number of M II oocytes as recipients. (iii) Lower oxygen tension and higher oxygen tension in IVM have no significant effect on development of cloned embryos. PMID- 16704121 TI - Study on genetic coadaptability of wild quail populations in China. AB - Genetic coadaptability of wild Japanese quail, wild Common quail and Domestic quail populations in China was studied using 7 microsatellite DNA markers and Monte Carlo method to test genetic disequilibrium. The molecular effects of genetic coadaptability were analyzed through a new statistical model of neutral site. The results showed that genetic coadaptability dominated the genetic disequilibrium of the three quail populations, and totally 16.67%, 9.66% and 10.05% of non-allelic combinations were in the genetic disequilibrium in wild Japanese quail, wild Common quail and Domestic quail populations, respectively. Genetic coadaptability existed at almost all the tested sites. In the molecular point of view, genetic coadaptability plays an important role of keeping lots of polymorphisms in natural populations. Therefore, it is another key factor to the genetic disequilibrium in the population except for linkage. The results enrich the conceptions and connotations of genetic disequilibrium, and help us know more about genetic coadaptability and its effects, and lay a foundation of evaluation and protection of wild quail genetic resources in China. PMID- 16704122 TI - Establishment of the model of white blood cell membrane chromatography and screening of antagonizing TLR4 receptor component from Atractylodes macrocephala Koidz. AB - A model of white blood cell membrane chromatography (WB-CMC) was established to screen active component from Atractylodes macrocephala Koidz. The component can antagonize Toll-like receptor 4 (TLR4) and inhibit inflammatory reaction. In the model of WB-CMC, cell membrane stationary phase (CMSP) was prepared by immobilizing the rabbit white blood cell membrane (WBCM) onto the surface of silica carrier and taxinol was used as a model molecule. The active component which can act on WBCM and its receptor (such as TLR4) as an effective target in A. macrocephala was determined by using a replacement experiment. The anti inflammatory effects of the active component were tested by using pharmacological methods in vivo. The results indicated that the retention characteristics of atractylenolide I as active component was similar to that of taxinol in the model of WB-CMC. And so, atractylenolide I acted on the WBCM and TLR4 and its anti inflammatory activity was related with antagonizing TLR4. Therefore, the interaction between the active component and WBCM and its receptor can be simulated by the model of WB-CMC in vitro. This model can be used to screen active components and to study effective characteristics for acting on definite targets. PMID- 16704123 TI - A novel model-free approach for reconstruction of time-delayed gene regulatory networks. AB - Reconstruction of genetic networks is one of the key scientific challenges in functional genomics. This paper describes a novel approach for addressing the regulatory dependencies between genes whose activities can be delayed by multiple units of time. The aim of the proposed approach termed TdGRN (time-delayed gene regulatory networking) is to reversely engineer the dynamic mechanisms of gene regulations, which is realized by identifying the time-delayed gene regulations through supervised decision-tree analysis of the newly designed time-delayed gene expression matrix, derived from the original time-series microarray data. A permutation technique is used to determine the statistical classification threshold of a tree, from which a gene regulatory rule(s) is extracted. The proposed TdGRN is a model-free approach that attempts to learn the underlying regulatory rules without relying on any model assumptions. Compared with model based approaches, it has several significant advantages: it requires neither any arbitrary threshold for discretization of gene transcriptional values nor the definition of the number of regulators (k). We have applied this novel method to the publicly available data for budding yeast cell cycling. The numerical results demonstrate that most of the identified time-delayed gene regulations have current biological knowledge supports. PMID- 16704124 TI - Pay for performance or performance for pay. PMID- 16704125 TI - Too much medicine? PMID- 16704126 TI - Beginning in error. PMID- 16704127 TI - Physicians health programs an international movement. PMID- 16704128 TI - Legal rights in transplantation: how adult stem cells may be defining and protecting the rights of donors. PMID- 16704129 TI - Medicare Health Support aims to change participant behavior, improve compliance. PMID- 16704130 TI - Hypertension prevalence awareness, treatment and control in North Nashville. AB - North Nashville is a largely African-American community known to suffer from increased cardiovascular mortality compared with whites in the same area of Davidson County. The burden of hypertension, a well-known risk factor for cardiovascular death, has not been fully described for this population. To quantify the prevalence, awareness, treatment and control of hypertension in the North Nashville community, we screened 132 patients aged 18 and older at the 2005 Juneteenth festival on Jefferson Street. We compared our results to data for African-Americans from the most recent NHANES survey. Compared to the national survey, our study population had a higher prevalence of hypertension (52.3 percent versus 33.5 percent) and lower awareness (68.1 percent versus 73.9 percent), treatment (46.4 percent versus 63.0 percent) and control of hypertension (13.0 percent versus 28.1 percent) despite comparable age. This study adds to a growing literature on the racial and geographic disparities in health status in the Unites States and cautions that data from national surveys may not be representative of individual communities. PMID- 16704131 TI - Intrauterine contraception: it's not what you think. AB - Our daughters are discovering intrauterine contraception. As their healthcare providers, we are in a position to educate them as they come to the office with questions. We should be prepared to give them current and accurate information. PMID- 16704132 TI - Secondary traumatic stress in mental health professionals. PMID- 16704133 TI - A botanical medicine, 1836. PMID- 16704134 TI - The OPTN/UNOS renal transplant registry. AB - This chapter summarizes analyses of 66,843 kidney-only transplants reported to the OPTN/UNOS Renal Transplant Registry between January 1999-December 2003. The 5 and projected 10-year graft survival rates for 28,260 recipients of living donor (LD) kidneys were 80.5% and 67.0%, respectively, based on a computed halflife of 17.8 years for those grafts that survived one year. Comparable results for 33,118 recipients of standard criteria deceased donor (SCD) and 5,943 recipients of expanded criteria deceased donor (ECD) kidneys were 68.8% and 50.9%, and 51.8% and 32.9%, respectively. The half-lives for SCD and ECD were 10.8 and 6.8 years, respectively. ECD kidneys (from donors aged 60+ or 51-59 with other risk factors) were preferentially transplanted to older patients (median age 58) even though there was no formal requirement to allocate these kidneys to older patients. The oldest recipients of deceased donor (DD) kidneys, 991 aged 71-75 and 254 over age 75 had 5-year graft survival rates of 48.9% and 40.7%, respectively and 40% of graft losses in these groups were due to death with a functioning graft. Among 469 LD kidney recipients over age 70, graft survival rates were 70% or higher with 40% of grafts lost due death with function. About 13% of DD and 5% of LD kidneys were transplanted to broadly sensitized patients during this period. One fourth and one-fifth of broadly sensitized patients who were transplanted received an HLA-matched DD or LD kidney, respectively. Overall, the proportion of transplants to unsensitized patients has increased in recent years to more than 50% of DD and nearly 70% of LD kidneys. There was no apparent correlation between the sensitivity of crossmatch tests and graft survival of first or repeat DD transplants; however, use of the more sensitive flow cytometry and AHG enhanced crossmatches significantly reduced the incidence of early rejections. Mean cold ischemia times have declined by about 7 hours between 1991- 2003, but the reduction in average cold time has not produced any notable reduction in the incidence of delayed graft function for recipients of SCD or ECD kidneys. Histocompatibility laboratories that waited for donor spleen or lymph node cells to perform final crossmatches added an average of 3 hours of cold ischemia for the kidneys they tested compared with laboratories that performed the tests with peripheral blood lymphocytes. There was no net increase in delayed graft function (DGF) with SCD kidneys tested at labs using spleen or nodes but the incidence of DGF among ECD kidneys was 25% when the lab used PBLs and 35% when the lab used spleen or nodes. PMID- 16704135 TI - Current status of thoracic organ transplantation and allocation in the United States. AB - The new Lung Allocation Score (LAS) system was created to address issues related to extended candidate waiting time, limited organ supply, waitlist mortality, and post-transplant survival. All candidates age 12 and older will be assigned an LAS that determines their priority for receiving lung offers. Waiting time will no longer play a factor in allocating organs to these candidates. Pediatric candidates under age 12 will continue to be allocated donor lungs based primarily on their accumulated waiting time. The LAS is the end product of an algorithm that assigns priority to lung candidates who are at higher risk of death if they do not receive a transplant (waitlist urgency) and who are likely to receive a greater benefit of longer lifetime with a transplant than without a transplant (transplant benefit). A variety of clinical factors, including disease diagnosis, were found to be predictive of waitlist urgency and post-transplant survival among transplant candidates, and are used in the calculation of each candidate's LAS. Pediatric and adolescent lung transplant candidates will have first priority for receiving offers from pediatric and adolescent lung donors. The lung allocation system will undergo frequent periodic reviews by OPTN/UNOS Committees to ensure that the performance goals of the system are being met and to address modifications to the system as they become necessary. The heart allocation system is an urgency status system under which adult and pediatric candidates are prioritized for organ offers by increasing levels of medical urgency. Median waiting time to transplant is shortest in the more medically urgent categories: Adult Status 1A=52 days, 1B=82 days, 2=468 days; Pediatric Status 1A=53 days, 1B=61 days, 2=149 days. Mortality among adult candidates after listing is strongly influenced by the medical urgency status at listing. Similarly mortality after transplantation for adult recipients also appears to be affected by the medical urgency status at transplant. PMID- 16704136 TI - The OPTN waiting list, 1988-2003. AB - Additions to the OPTN waiting list decreased in 2003 for all organs except kidney. On November 30, 2004 there were 95,598 registrations on the combined UNOS waiting list. Of these, 67% were awaiting kidney transplantation and 18% were awaiting liver transplantation. The majority of patients on the UNOS waiting list on November 30, 2004 were blood type O (51%), male (57%) and awaiting their first transplant (87%). Despite lengthy waiting times, the percentage transplanted within one year following listing increased over the past 3 years for all organs except heart and kidney. A substantial increase in the percentage of heart-lung candidates transplanted within one year was observed in 2003. Blood type and medical urgency had a significant impact upon percent transplanted within one year of listing for most organ types. Patients awaiting heart, liver, pancreas and intestine transplants had the highest probability of receiving a transplant within one year. Death rates have remained fairly stable over time; however, cause of death while waiting for a transplant varied with organ type. Death due to cardiovascular reasons was the most common cause of death for heart and kidney candidates who were on the UNOS waiting list, while multiple organ failure was the most common cause of death for liver and intestinal candidates who were on the UNOS waiting list. PMID- 16704137 TI - Organ donation in the United States: 1988-2003. AB - There were 6,457 deceased and 6,821 living donors recovered in 2003, a 58% and 274% increase over those recovered in 1988. The number of deceased donors aged 50 or older has increased from 12% of all donors in 1988 to 34% of all donors in 2003. The typical deceased donor in 2003 was a white male with ABO blood type O between the ages of 35-49. In 2003, a typical living donor was a white female with ABO blood type O between the ages of 35-49. Between 1988 and 2003, the percentage of minority donors increased for deceased donors (17% to 30%), and for living donors (24% to 30%). The number of living donors who were either spouses or unrelated to the recipient increased from 5% in 1988 to 33% in 2003. In 2003, California (10.2%) was most often listed as the state of residence for deceased donors, followed by Texas (7.9%) and Florida (6.9%). In 2003, deceased donors were recovered most often on Wednesdays (16.0%), followed by Tuesdays (15.2%) and Thursdays (14.7%). In 2003, living donors were recovered most often on Tuesdays (26.4%), followed by Wednesdays (25.5%) and Thursdays (20.7%). In 2003, deceased donors were recovered most often in October (9.0%), followed by January (8.6%), and July (8.6%). In 2003, living donors are recovered most often in July (10.2%), followed by October (8.9%) and June (8.6%). PMID- 16704138 TI - Outcomes associated with recipient and donor hepatitis C serology status after kidney transplantation in the United States: analysis of the USRDS/UNOS database. AB - Analysis of the USRDS kidney transplant registry disclosed that use of hepatitis C virus-positive donor (DHCV+) kidneys was an independent risk factor for patient death after kidney transplantation when compared to use of DHCV- kidneys, and that death in recipients of DCHV+ kidneys occurred earlier than previously reported. This increased risk of death was delayed for about 2 years, suggesting the development of an intermediate complication that resulted in a later increased risk of death. While liver disease early after transplant in these patients was rare, new-onset diabetes mellitus occurred early and commonly, suggesting this complication as a prominent mediator of mortality associated with transplantation with DHCV+ kidneys. Identification of new-onset diabetes mellitus may represent a new target of opportunity to improve outcomes associated with use of DHCV+ kidneys. Even under the current circumstances, use of DHCV+ kidneys was also independently associated with a survival experience that, although less favorable than associated with transplantation of DHCV- kidneys, was significantly better than remaining on the kidney transplant waiting list. Whether this survival advantage applies to all relevant subgroups could not be assessed and warrants further study. Our analyses suggest opportunities to improve survival and reduce morbidity after use of DHCV+ kidneys. PMID- 16704139 TI - Review of transplantation in HIV patients during the HAART era. AB - Human immunodeficiency virus (HIV) infection was once thought to be a relative or even absolute contraindication to transplantation. With the recent advent of highly active antiretroviral therapy (HAART), those infected with HIV are now living longer and dying from illnesses other than acquired immunodeficiency syndrome (AIDS). Although studies prior to the HAART era suggested poor outcomes might occur with transplantation in those infected with HIV, more recent studies have demonstrated results comparable to those of recipients without HIV infection. A number of issues persist regarding ethics, patient selection, post operative management, and drug interactions between antiretroviral and immunosuppression agents. In this review, kidney, liver, and heart transplantation in the HIV-positive population were analyzed using data from the Organ Procurement and Transplantation Network/ Scientific Registry of Transplant Recipients. PMID- 16704140 TI - The role of an organ exchange organization in increasing split-liver transplantation. AB - Eurotransplant introduced a new allocation policy in January 2003 to increase the number of liver transplants by offering centers an incentive to split deceased donor livers for 2 recipients. Centers were granted the option of choosing a suitable recipient for the second portion of the split liver from their own waiting list and, to increase the awareness for liver splitting, centers were asked by the Eurotransplant duty officer whether they would consider splitting whenever a liver that met the 50/50 rule (donor age <50 and weight >50 kg) was available. During the first year after implementing this policy, split-liver transplants increased by 67% and again by 10% during the second year (a total of 288 transplants in the 2-year period). The number of pediatric recipients of a split liver increased from 44 in 2002 to 76 in 2004 and the pediatric waiting list decreased by 36% (73 to 47) one year after implementation of the new policy. More than 95% of the 288 split liver transplants involved one adult and one pediatric recipient. Nearly three-quarters of the split liver transplants were performed at 3 centers with both a pediatric and adult waiting list and with surgeons experienced in the procedure. We conclude that Eurotransplant's liver allocation policy has increased the number of liver transplants, particularly among children, by rewarding centers that split livers for transplantation to 2 recipients without prolonging cold ischemia time. The number of centers that could benefit from this policy will increase as more surgeons are trained in the splitting procedure. PMID- 16704141 TI - Outcomes of kidney transplants from non-heart-beating deceased donors as reported to the Japan Organ Transplant Network from April 1995-December 2003: a multi center report. AB - Between April 1995-December 2003, 1,324 deceased donor kidney transplantations were performed in 139 transplant institutes in Japan. Of these, 45 transplants were from heart-beating and 1,279 transplants were from non-heart-beating deceased donors (NHBDD). Clinical outcomes for the 1,279 recipients of NHBDD kidney transplants were investigated. The overall 5-year patient and graft survival rates were 90% and 72%, respectively. A total of 112 NHBDD kidney grafts never functioned after transplantation and the recipients had to remain on dialysis. The causes of nonfunction were rejection, primary nonfunction, death, thrombosis and others in the order of the incidence. The major causes of graft loss were nonfunction, death, chronic rejection and acute rejection in that order. Major causes of recipient deaths were pneumonia, sepsis and CVA within 12 months, and heart diseases, sepsis, malignancy and pneumonia more than 12 months after transplantation. Kidneys from female donors, donors aged 15 or less or over age 60, donors with extrinsic causes of death other than head trauma, recipients over age 60 and those with diabetic nephropathy as their original disease were found to be at risk for poor graft survival. The lowest and last donor serum creatinine level did not influence the incidence of nonfunction or graft survival. However, graft survival was significantly poorer among recipients of older "expanded" donor kidneys than for recipients of younger grafts. The warm and total ischemia times should be kept shorter than 30 minutes (better 15 minutes), and 12 hours, respectively to minimize the incidence of nonfunction and early graft loss. It is especially important in cases with WIT over 30 minutes that the total ischemia should be kept within 12 hours. Cannulation before cardiac standstill was important to reduce the incidence of nonfunction and achieve high graft survival rates with NHBDD kidneys. The discontinuance of ventilator support also reduced the incidence of graft nonfunction. The combination of CsA or Tacrolimus and MMF as both the induction and maintenance regimen significantly improved graft survival. The use of either anti-T cell antibodies or basiliximab was also associated with significantly better graft survival for NHBDD kidneys. The combination of basiliximab, CsA and MMF resulted in a graft survival rate of 98% at one and 2 years. PMID- 16704142 TI - Report from the National Transplantation Pregnancy Registry (NTPR): outcomes of pregnancy after transplantation. AB - The NTPR maintains an ongoing active database to study the safety of pregnancy in transplant recipients and currently includes the outcomes of more than 900 female recipients who became pregnant after their transplant and just over 700 male recipients who fathered one or more pregnancies after receiving a transplant. Analyses include the long-term follow-up of the recipient's graft status and their offspring. Successful pregnancy outcomes have been noted for each solid organ recipient group. The Registry includes information on 1,097 pregnancies in 716 kidney recipients, 187 pregnancies in 111 liver recipients, 56 pregnancies in 38 P/K recipients and smaller numbers for other organs and combinations of organs. There are periodic reports of recipients with graft dysfunction, rejection, or graft loss that may be related to pregnancy events, though the majority of outcomes reported to the NTPR appear favorable for parent and newborn. Organ-specific issues and comorbidities must also be considered in analyzing outcomes. The pregnancy issues that face recipients and caretakers with the current newer adjunctive therapies and newer immunosuppressive regimens require ongoing study. The potential risk of teratogenicity must be weighed against the potential risk of rejection when altering drug regimens before planned conceptions or in making dosage adjustments during pregnancy. Unplanned pregnancies present obvious concerns. Pregnancy safety has not been established for either MMF or sirolimus and all centers are encouraged to report pregnancies with exposures to these agents to the NTPR. Continuing analyses are directed at potential effects of the newer immunosuppressive regimens, not only to identify any risks to the pregnancy from immediate exposure, but also for potential postpartum exposures such as from breastfeeding. As the registry study design allows for continued contact, efforts continue to accrue long-term follow-up of both parent and child. PMID- 16704143 TI - Post-transplant parenthood. PMID- 16704144 TI - "High-risk" renal transplantation: evolving definitions at a single center. AB - Characterization of renal transplant recipients as "high-risk" originated in the 1970's, although attributes that defined this category, such as diabetes mellitus, are no longer applicable. The changing paradigm of risk after renal transplantation reflects the impact of nonspecific advances in clinical care, specific interventions that address previously defined problems, changing demographics, and new issues that have arisen as a consequence of changes in clinical practice. In the current era, diabetes, retransplantation, and presensitization are no longer considered risk factors for poor outcomes after kidney transplantation. Significant risk factors influencing intermediate-term graft survival now include donor and recipient age over 60 years, DR mismatching (only in kidneys from deceased donors), time awaiting transplantation, and African-American race. Current clinical approaches to care of the renal transplant candidate/recipient focus on minimizing the impact of identified risk factors rather than avoiding transplantation altogether. For most ESRD patients, the greatest risk lies in not receiving a transplant. PMID- 16704145 TI - Transplantation of A2 and A2B kidneys from deceased donors into B waiting list candidates increases their transplantation rate. AB - Transplant centers in the Midwest Transplant Network began transplanting kidneys from A2 or A2B donors into blood group B and O patients in 1986. Since 1991, an OPTN/UNOS variance has permitted us to allocate these kidneys preferentially into B and O waiting list patients. With more than 10 years of experience we have noted the following: 1. Thirty-one percent more blood group B patients were transplanted by allocating them A2 or A2B kidneys from our deceased donors. 2. Ten-year graft survival for B recipients of an A2 or A2B kidney (72%) was equivalent to that for B recipients of a B kidney (69%). 3. Type B recipients of simultaneous pancreas-kidney transplants (n=4) also did well with A2 or A2B organs. 4. Non-A recipients were transplanted only when their anti-A IgG titer history was consistently low (< or =4). 5. Most (90%) blood group B patients had a low anti-A IgG titer history; whereas, only one-third of blood group O patients had a low titer history. 6. Neither ethnicity nor HLA class I sensitization level influenced the anti-A IgG titer history. 7. In an OPO with mostly (87%) white donors, nearly 20% of blood group A donors were A2. 8. Waiting time until transplantation was lower for B patients who received an A2 or A2B kidney than for those who received a B or O kidney. 9. Our OPO blood group B waiting list was reduced from 25 low PRA (<40%) B candidates in 1994 to 4 in July, 2004. 10. Blood group A candidates received 6.4% fewer transplants with our A2/A2B--> B allocation algorithm. 11. Minority patients were transplanted at the same rate when using the A2/A2B--> B allocation algorithm as when using the standard UNOS algorithm for allocating B and O kidneys--> B patients. PMID- 16704146 TI - Clinical outcome and accommodation in ABO incompatible kidney transplantation. AB - We performed 84 ABO-incompatible kidney transplants at Toho University since 1989, with plasmapheresis and exchange replacing AB blood group plasma as pre conditioning to reduce anti-donor blood group antibodies. Our current immunosuppression protocol consists of basiliximab, MMF, steroid, and cyclosporine or tacrolimus, including splenectomy. Overall patient/ graft survival rates (n=84) were 95/93 at one year, 94/92 at 3 years, 87/80 at 5 years, 87/75 at 7 years, and 83/67 at 10 years. The outcomes are similar to those of ABO compatible living donor transplants. We have achieved 100% graft and patient survival rates (n=48) for the 7 years since January 1997. Our findings suggest that post-conditioning is not necessary to control titers of anti-donor blood group antibodies or to overcome acute humoral rejection. Infection control is critical in achieving good outcomes in ABO-incompatible transplants. We found that only anti-donor blood group antibodies in blood group O recipients of ABO incompatible kidneys were specifically suppressed one year after transplantation. This appeared to be a type of accommodation in which there was no immunological response despite the co-existence of donor antigen and antibody, and might have been caused by down-regulation of B cells to produce anti-donor antibody. PMID- 16704147 TI - Kidney transplantation at the University of Illinois at Chicago from 1988-2004. AB - The Division of Transplantation at the University of Illinois at Chicago (UIC) was established in 1968 by Dr. Olga M. Jonasson, who performed the first living related kidney transplant in Illinois. The UIC Hospital is a typical inner city hospital serving primarily minorities and under-privileged populations with significant challenge for issues such as compliance, insurance coverage, low income, and degree of education. We analyzed the outcomes of 67 pediatric and 801 adult consecutive renal allotransplantations performed at the University of Illinois at Chicago (UIC) from January 1988-December 2002 and more recent information in relation to newly developed programs (robotic living donation, transplantation in sensitized patients, early steroid discontinuation, ethnicity tailored immunosuppression) until November 2004 (21 pediatric and 172 adults). The one-year graft survival has improved comparing the cases performed between 1988-1998 versus cases after 1998 from 82-88%, with similar patient survival. However, the overall 5-year patient and graft survival has not changed significantly in the 2 different eras and is currently 85% and 65%, respectively. With the use of minimally invasive, robotic-assisted living donor nephrectomy, the use of living donors has increased and these now represent 75% of the kidney donors at UIC. Transplantation in highly sensitized recipients has become more successful with the use of plasmapheresis resulting in 100% patient and graft survival and a 10% acute rejection rate in the first series of 20 patients. Early steroid discontinuation (steroid discontinued at day 6 after transplantation) has allowed for significant improvement of results in pediatric recipients. We have 100% one-year patient and graft survival and no acute rejections in a series of 13 children, compared with a 25% rejection rate in 13 children with steroid containing immunosuppession. Tailored immunosuppression with early steroid discontinuation now achieves equal early results in African American as in any other ethnic group with 98-99% patient and graft survival and acute rejection rates below 10%. Of interest will be the future evolution of long-term patient and graft survival under the new immunosuppressive protocols with early steroid discontinuation achieving very low acute rejection rates and impressive one-year survival rates. There is reason for optimism that the improved early results will translate into better long-term outcomes. PMID- 16704148 TI - One thousand renal transplants in Belfast (1968-1998). AB - The first kidney transplant was performed in Belfast in 1968. During the next 30 years 1,000 transplants were undertaken at this unit. Data were analysed on 937 cadaveric transplants, 815 first and 122 regrafts. There were 63 living transplants. Long-term follow-up was achieved for all grafts except one live transplant. All recipients had follow-up of at least 5 years. One- and 5-year graft survival rates were 78.9% and 65.0% for first cadaveric grafts, 83.6% and 66.4% for regrafts and 85.7% and 68.3% for living transplants. Nine of 41 transplants performed more than 30 years ago are still functioning. Multivariate analysis determined risk factors for graft survival as recipient age, donor age, HLA-A mismatching, HLA cytotoxic antibody level and year of transplant. A conservative regime regarding the use of immunosuppression, HLA matching and crossmatching has proved successful in accomplishing good graft survival at this unit. PMID- 16704149 TI - Present status of pancreas transplantation in Japan. AB - Since October 1, 1999, a total of 17 pancreas transplantations have been performed in Japan, involving 15 brain-dead donors and 2 non-heart beating donors. The donor ages were relatively high with 67% (10/15) over age 40 and brain death was typically attributable to cerebrovascular accidents (70.6%; 12/17). The majority of donors could be defined as marginal (13/17; 76.5%). Because of these adverse donor characteristics, the gastroduodenal artery of the graft was usually reconstructed to increase blood supply to the pancreas head when liver procurement was undertaken. Also, intensive insulin therapy was used to rest graft islets in some centers during the acute phase of transplantation. Patient survival was 100% with a mean follow-up period of 21 months (range: 4-54 months). All recipients have remained insulin-free without any need for dialysis. One patient lost a pancreatic graft to venous thrombosis, but was successfully retransplanted 2 years later. All recipients have displayed good QOL after transplantation. Nine recipients made a complete return to their jobs, while 5 patients made a partial return to light jobs, such as housework or office work. PMID- 16704150 TI - Twenty years of clinical islet transplantation at the Diabetes Research Institute -University of Miami. AB - Transplantation of allogeneic pancreatic islets for the treatment of patients with Type 1 diabetes mellitus (T1DM) is now a reality. The steady progress that has allowed for the recent successful clinical trials world-wide follows a steep learning curve and the perseverance of the international islet transplantation community. The Clinical Islet Transplant Program at the Diabetes Research Institute - University of Miami has contributed to the progress in the field with a 20-year track record. It has been a long journey and despite the intermediate success, more work is needed in order to achieve the ultimate goal of a safe and long-lasting treatment for patients with T1DM. PMID- 16704151 TI - Pancreatic transplantation at the University of Pittsburgh. AB - Campath-1H preconditioning with tacrolimus monotherapy is an effective immunosuppressive regimen for pancreas transplantation, with acceptable patient and graft survival rates early after transplantation. Rejection rates are low under this protocol if the tacrolimus level is kept consistently >10 ng/ml. This immunosuppressive protocol, combined with recent technical refinements, has resulted in lower rates of thrombosis and overall complications. Pancreatic transplantation en-bloc with visceral grafts has the following unique features: Diabetes is a rare indication, and HLA matching is not required. The gland is immunologically protected by the simultaneously transplanted visceral organs. Disease gravity, surgical complexity and gut alloimmunity influence the overall pancreatic allograft survival. The current UNOS listing criteria and data registry should be modified for obvious logistic and scientific reasons. PMID- 16704152 TI - CNI-sparing regimens within the liver transplant setting: experiences of a single center. AB - One- and 5-year survival rates after liver transplantation have improved with the development of potent immunosuppressive drugs. Thus, attempts have shifted to reduce long-term complications of CNI and prednisone therapy. Minimization of CNI and concomitant introduction of MMF has resulted in a significant improvement of renal function, blood pressure and triglyceride values in our transplant patients. In recipients who develop recurrent or de novo malignancies, SRL therapy might be recommended. In addition, the flexibility to individualize immunosuppression should be favored in selected patients. However, large prospective randomized trials are required in order to determine if SRL/low dose CNI or SRL monotherapy are reasonable options for prolonging patient survival. Moreover, important questions such as the appropriate SRL target dose for prevention of tumor growth need to be answered. Numerous studies investigating distinct immunosuppressive combination therapies and withdrawal or minimization of CNI are available in the literature; however it is not yet clear which treatment regimens will prove to be the most efficacious and least toxic. PMID- 16704153 TI - The Baylor Regional Transplant Institute: review of a twenty-year experience. AB - As the Baylor Regional Transplant Institute celebrates its 20-year anniversary and the transplant of its 2,500th liver, the program has continued to grow, reaching its highest yearly volume to date. Twenty years gives an excellent opportunity to see how the facets of our program have adjusted in response to changes in both society and our own field of transplantation. Our recipient and donor demographics have changed significantly, but despite older recipients, older and more marginal donors and the issues of disease recurrence, we have consistently improved survival of both the patient and graft, reduced rejection rates and minimized hospital and ICU stays. These survival improvements have been even more pronounced in patients transplanted for HCC. Because of constantly changing risk and complication patterns, we have continually analyzed and adapted our processes and protocols, adjusting them in response to patient outcomes and the best current scientific evidence when needed. The research at Baylor has always had a practical, clinical dimension, but with development of our "Genes of Health" program at the Baylor Institute of Immunology Research, we hope to expand more basic science transplant research and most importantly take the lead in translational research. PMID- 16704154 TI - Cardiac transplantation at Duke University Medical Center. AB - Since beginning cardiac transplantation in 1985, Duke University Medical Center has performed 485 de novo heart transplants in adult recipients. Our program has broadened the access of patients to transplantation through the aggressive use of ventricular assist devices as bridge to transplant in patients with acute heart failure and shock, as well as through the introduction of an alternate list program for suboptimal transplant candidates. Despite this broadening of the recipient pool, our long-term outcomes have remained good, with 5-year survival of 75% and 10-year survival of 51%. Continued advances in immunosuppression and treatment for cardiac allograft vasculopathy seem likely to continue to improve long-term outcomes from cardiac transplantation. Expanding indications for VAD support (such as destination therapy) and continued improvements in this technology seem certain to impact patient selection and therefore the results of transplantation in the future. Appropriate triage of patients with advanced heart failure among available therapies will be the cornerstone of optimizing outcomes in this critically ill patient population. PMID- 16704155 TI - Crossing the ABO barrier in infant heart transplantation at the Hospital for Sick Children. AB - Heart transplantation in infancy is generally associated with excellent clinical results. Unfortunately, heart transplantation cannot be offered to many infants who would benefit from this therapy due to a shortage of organ donors of suitable size for infants, and traditional limitations that constrain donor availability still further, such as the requirement for ABO-compatibility. At the Hospital for Sick Children, we questioned the need to apply this requirement to infants, based on available scientific evidence regarding the immaturity of certain immune responses. Our experience with 23 ABO-incompatible infant heart transplants demonstrates that this procedure can be performed safely in infants. Moreover, we have shown that donor-specific B-cell tolerance develops following ABO incompatible transplantation. The consistency of the clinical and laboratory outcomes of ABO-incompatible infant heart transplantation to date would suggest that there is no scientific rationale to require ABO-compatibility between donors and recipients for infant transplantation. PMID- 16704156 TI - Proteomics in renal transplantation: opportunities and challenges. AB - Proteomics is a fast growing scientific field that has been developed during the past 10 years. The central goals are to define the protein composition of complex biological samples, to explore alterations in protein expression related to various processes/diseases and to study functional aspects of protein-protein interactions. To date, proteomic studies are rare in renal transplantation; however, they may enhance our understanding of transplant pathophysiology and help to identify biomarkers and novel targets for rational drug therapies. The purpose of this review is to describe the possibilities and limitations of differential expression proteomic technologies with a focus on biomarker development for non-invasive renal allograft monitoring. PMID- 16704157 TI - Microarrays: interrogating the transplant transcriptosome. AB - The use of DNA microarrays as a hypothesis generation tool for determining gene expression differences across thousands of genes in a data set is increasing. Recently, the use of microarrays has been applied to the transplant field and holds great promise for unraveling the mechanisms at play in various transplant processes and for identifying new tissue-specific and noninvasive biomarkers predictive of clinical outcomes. As microarrays produce large amounts of data, bioinformatics tools are being developed to determine gene expression patterns. Gene clustering and class prediction tools aid in the discovery of molecular signatures in different disease processes while literature mining, gene family analysis and pathway analysis helps in understanding the biological relevance of these signatures. This chapter focuses on DNA microarrays, their application to transplantation, and reviews key research studies where DNA microarrays are applied to understand acute rejection, chronic rejection and tolerance in human transplantation. PMID- 16704158 TI - The year in review--2004. PMID- 16704159 TI - Explainable variation in renal transplant outcomes: a comparison of standard and expanded criteria donors. AB - In 2002, OPTN/UNOS altered kidney allocation rules to allow patients to be listed separately to receive kidneys from expanded criteria donors (ECD). Our aim was to quantify the short- and long-term impacts of 21 prognostic factors on recipients of ECD as well as recipients of living (LD) and deceased standard criteria (SCD) donors. A factor's impact depends on both the risk and diversity of its effects. Using OPTN/UNOS Registry data from 1996-2003, we have analyzed kidney-only, adult recipient grafts for factor effects among 35,878 LD, 47,941 SCD and 10,399 ECD transplants. During an early risk period, all 94,218 recipients were followed through one year, and, in the late risk period, 85,270 recipients whose grafts survived beyond one year were followed for 5 years post-transplant. Impact was measured by determining a factor's percentage of assignable variation in one- and 5-year graft failure rates. Scores for 21 factors were estimated via generalized logistic models, which contained a random component for transplant center. The assignable variation associated with a given factor was computed as the factor score variance multiplied by the square of the corresponding regression coefficient. Impacts were heterogeneous with regard to posttransplant period and donor type. The top 5 factors influencing one-year graft survival rates were as follows: * For LD grafts - pretransplant dialysis time (14% of the variation in short-term outcomes), recipient age (13%), body mass (12%), PRA (10%) and induction therapy (10%). * For SCD grafts - donor age (24%), recipient age (12%), pretransplant dialysis time (12%), HLA-DR matching (6%) and pretransplant medical condition (6%). * For ECD grafts - donor age (18%), pre-transplant dialysis time (10%), recipient age (10%), pretransplant medical condition (10%) and recipient body mass (6%). Ranking long-term outcomes demonstrated the following top 5 influential factors: * For LD grafts - donor age (28% of the variation in long term outcomes), recipient race (15%), age (15%), transplant year (13%) and recipient sex (11%). * For SCD grafts - donor age (35%), recipient race (23%), transplant year(15%), recipient sex (8%)and age (5%). * For ECD grafts - donor age (33%), recipient sex (20%), race (15%), transplant year (8%) and recipient's original disease (5%). Donor age was the dominant factor governing the survival rates among deceased donor kidney transplants. Advancing donor age was still the major risk factor for SCD transplant failure despite setting aside all donors 60 and up, and a large fraction of 50-59 year-old donors, from this group. Current ECD/SCD definitions warrant review and possible revision. PMID- 16704160 TI - An analysis of the OPTN/UNOS Liver Transplant Registry. AB - Based on data reported to the OPTN/UNOS Liver Transplant Registry between 1988 2004: 1. There was a very small difference in 5-year graft survival rates comparing living and deceased donors in adult (4.3%) and pediatric patients (2.4%). 2. Although graft survival rates of split liver transplants were lower than whole liver grafts before 1998, 5-year graft survival results of more recent split grafts (65.8%) have become comparable to those of whole liver grafts (66.5%). Among recipients in good condition, split (67.7%) and whole grafts (70.0%) yielded equivalent survival rates. 3. Lower graft survival rates were noted in ABO incompatible grafts, non-heartbeating donors, regrafted patients, and recipients who were in the ICU before transplantation. 4. There was no recipient gender effect on liver transplant outcome. 5. Primary disease distributions were different for different races. Among adult patients, the largest fraction of white patients had alcoholic cirrhosis. Among Asians, Type B cirrhosis was most frequent. Among pediatric patients, biliary atresia constituted the majority of patients. Most of the patients with alpha-1 antitrypsin deficiency were white. Autoimmune hepatitis was most frequently found among black patients. 6. Although 5-year graft survival of black patients (60.2%) was lower than whites (68.1%), Hispanics (67.6%), and Asians (68.0%), black recipients with PBC (73.3%) and PSC (69.9%) had graft survival rates similar to those of whites (78.1%) (73.6%) and Hispanics (75.3%) (77.1%). 7. Zero HLA-A,-B, DR mismatched livers had very rapid early failures. HLA matching correlated with graft survival in autoimmune hepatitis patients, but not in cirrhosis patients. 8. Short-term graft survival for liver transplants has improved steadily since 1990. However, long-term graft survival after the first year actually declined over time. 9. In adult transplants, 5-year graft survival of autoimmune-related diseases, PBC (77.3%), PSC (73.3%), AIH (74.2%) yielded higher graft survival rates than those of hepatitis B (71.5%) and C (63.2%). 10. In pediatric patients, 5-year survival of biliary atresia (75.4%), autoimmune cirrhosis (70.8%), and alpha-1-antitrypsin deficiency (85.0%) had high graft survival rates, except for acute liver failure (61.6%). 11. Hepatitis C recurrence is now one of the major causes of graft failure in adults. Thrombosis is a major factor in graft failure for pediatric transplants. PMID- 16704161 TI - Heart transplantation in the United States 2004. AB - The limited availability of donated hearts is not sufficient to meet the needs of increasing numbers of heart transplant candidates. Thus, it is important to find the best recipient for each donated heart in order to achieve the best graft and patient survival. We investigated heart transplant outcomes in the US and the factors that influence them. We present information regarding 36,277 heart transplants reported to the OPTN/ UNOS Registry from January 1988-October 2004. The annual number of transplants ranged between 2,057-2,363 during the past 14 years and has not increased since 1990. One- and 5-year graft survival rates have gradually increased and have reached 87% and 72%, respectively, for adult recipients. Non-immunological factors, such as the circumstance of the donor's death, cause of death and ischemic time, the recipient's race, age, original disease, infection, systemic cerebro-vascular disease, whether or not they were on life support, renal function etc, significantly impacted on graft survival. Immunological factors, such as previous transfusion, previous transplantation, HLA mismatches, sensitization status, rejection episodes before discharge or within one year after transplant, were associated with lower graft survival rates. In conclusion, in order to achieve the best possible outcome for heart transplant patients, we must first find the most suitable candidate for an available donated heart; secondly, we must optimize immunosuppressive regimens and other medical and surgical therapies to prevent graft rejection and transplant-related complications. PMID- 16704162 TI - Serum creatinine as a predictor of transplant survival and death. AB - Taking ALL patients transplanted in a given calendar year, we have shown that serum creatinine values have gradually decreased over the past 15 years. Donor age was a major factor influencing serum creatinine. This was followed by the recipient sex. Of lesser importance was the different immunosuppressive drugs employed, and whether the donor was deceased or living. The strong predictive value of serum creatinine was shown at various times after transplantation. Also, one year prior to failure, the creatinine values were very significantly elevated. A similar finding was noted one year prior to the death of patients with a functioning graft. PMID- 16704163 TI - Impact of diabetes mellitus on transplantation. AB - 1. In total, 99,933 kidney alone transplants were performed in the US from 1995 2002. Of these, 11,251 transplants were for type 1 diabetic recipients, 7,772 transplants were for type 2 diabetic recipients and 80,910 for non-diabetic recipients. Approximately 20% of all kidney transplants were performed for diabetic recipients during this period. 2. The mean age of recipients and donors has been increasing annually since 1995 for type 1 and type 2 diabetic recipients. The same trend was noted for the mean BMI of recipients. 3. Kaplan Meier estimates of kidney transplant survival for each variable showed that the age of recipients and donors, BMI of recipients, recipients' race, HLA-DR3/4 positive of the recipients, types of primary payment source, presensitization of the recipients, donor type, number of HLA mismatches, and cold ischemia time were significantly associated with the patient and kidney graft survival rates for type 1 diabetic recipients. 4. When adjusted for 15 potential risk factors in the Cox proportional hazard model, recipient age and sex, source of primary payment, donor age, donor type, and the number of HLA mismatches were independent and significant predictors of patient survival. With respect to kidney graft survival, recipient age and race, source of primary payment, peak value of percent panel-reactive antibodies, donor age, donor type, and the number of HLA mismatches were independent and significant predictors. 5. Kaplan-Meier estimates of the entire survival of a kidney transplant for each variable showed that recipient and donor age, types of primary payment source, donor type, number of HLA mismatches, and cold ischemia time were significantly associated with kidney graft survivals for type 2 diabetic recipients. 6. When adjusted for 15 potential risk factors in the Cox proportional hazard model, recipient age and BMI, source of primary payment, donor age, and donor type were independent and significant predictors of patient survival. With respect to kidney graft survival, recipient age, and BMI, source of primary payment, donor age, donor type, and number of HLA mismatches were independent and significant predictors. 7. In conclusion, our data suggests that recent kidney transplants have been performed among older diabetic patients with higher risk. Further studies, considering all variables implicated in the prognosis of diabetes and its impact on graft and patient survival are needed in order to improve the outcome of diabetic recipients after kidney transplantation. This may also help us optimize the use of limited kidney grafts. PMID- 16704164 TI - Death with a functioning graft in kidney transplant recipients. AB - The percentage of deceased donor kidney transplant recipients who died with a functioning graft (DWF) within one year after transplantation increased by 1% between 1993-1995, while DWF between 2-5 years after transplant has decreased since 1992, after a 3-year of increase (1989-1992). Deaths due to infection or hemorrhage within the first post-transplant year have increased since 1995 and deaths from cardiovascular disease between 2-5 years after transplantation have decreased. Deaths from malignancies among recipients of living donor kidneys increased significantly from 0.5% to 0.7% after 1995. Diabetic recipients (type II) of deceased donor grafts had a higher incidence of DWF (5% in the first year and nearly 10% between 2-5 years) than recipients with other diseases. Among diabetic recipients of living donor kidneys, DWF occurred in 3% during the first year and 7% between 2-5 years. The incidence of DWF increased as the recipients age increased, from less than 1% among recipients under age 18 to more than 10% of patients over age 60. A multivariable analyses of deceased donor transplants identified the donor age (>45), recipient age (46-60, >60), diabetes, CMV status and intermediate PRA as significant risk factors for DWF. After transplantation, DGF and certain immunosuppression regimens at discharge also influenced DWF. The multivariable analysis results for living donor transplants were similar, except that donor factors had no significant effect on DWF. PMID- 16704165 TI - Pathology of B-cell non-Hodgkin's lymphomas and multiple myeloma. PMID- 16704166 TI - Pathogenesis of viral lymphomas. PMID- 16704167 TI - The indolent B-cell lymphomas. PMID- 16704168 TI - Diffuse large B-cell NHL. PMID- 16704169 TI - Mantle cell lymphoma: current concept in biology and treatment. PMID- 16704170 TI - T-cell non-Hogdkin's lymphoma. PMID- 16704171 TI - Monoclonal antibodies in the treatment of non-Hodgkin's lymphoma. PMID- 16704172 TI - Blood and bone marrow transplantation for patients with Hodgkin's and non Hodgkin's lymphoma. PMID- 16704173 TI - Vaccine therapies for non-Hodgkin's lymphomas. PMID- 16704174 TI - Management of early stage Hodgkin's lymphoma. PMID- 16704175 TI - Management of advanced stage Hodgkin's lymphoma. PMID- 16704176 TI - Nodular lymphocyte predominant Hodgkin's disease. PMID- 16704177 TI - Lymphoma imaging: nuclear medicine. PMID- 16704178 TI - Novel small molecules in the treatment of lymphomas. PMID- 16704179 TI - Cardiac transplantation in South Carolina: 300 transplants. AB - For nearly 20 years, the Medical University's Heart Transplant Program has been providing the citizens of South Carolina with excellent results with a minimum of delay. We present here the results of our first 300 heart transplants, spanning the first 18 years of the Cardiac Transplant Program at the Medical University. Overall survival has been very good, with one, five and ten year survival rates in the adults being 92 +/- 2%, 78 +/- 3%, and 58 +/- 4%. The children's group showed survival rates of 94 +/- 5%, 79 +/- 11%, and 79 +/- 11% over the same lengths of time. Most recently, the federally sponsored Scientific Registry of Transplant Recipients (www.ustransplant.org, July 2005) reports for MUSC a one year survival of 97.67% and three-year survival of 90.74%; both leading the Southeast. We attribute this success to the dedicated work of health care workers at all levels who believe in attention to detail and that the patient always comes first. It is our hope that we will be able to continue to provide expert, state-of-the-art, cardiac transplant services long into the future, while continuing to expand our heart failure management program as dictated by further developments in this rapidly evolving specialty. PMID- 16704180 TI - The gaping hole: physicians are missing from the front line of disaster preparedness training. PMID- 16704181 TI - Cystic fibrosis screening at the Greenville Hospital System. AB - The cost effectiveness of prenatal screening for cystic fibrosis needs to be reexamined. This will require further study. At the time prenatal screening for CF carrier status was initiated, newborn screening for cystic fibrosis was not commonly available. The benefit of screening in our population was detection of 68 female carriers; however, this did not result in finding any fetuses affected with cystic fibrosis. The American College of Obstetrics and Gynecology has recommended CF screening for women of childbearing age but as yet no consensus has been reached concerning the cost effectiveness of this policy. Although over 2600 pregnancies were tested, the reproductive outcome was not altered in a single pregnancy. PMID- 16704183 TI - Orthopaedic surgery: observations of a medical student. PMID- 16704182 TI - Your patients may be at risk: preventing pediatric burn injuries. AB - Burn injuries remain a leading cause of unintentional injuries in children. Improvements in burn care technology, comprehensive outpatient services, and a team approach to care can reduce hospital stays and ease recovery for patients and families. Burn awareness and fire prevention education to the community and to health care providers may further decrease the incidence of pediatric burn injuries. Primary care providers can have a great impact on reducing the opportunity for a burn injury in a child by sharing what is known about burn incidence and prevention with the children and families they serve. PMID- 16704184 TI - The seven basic virtues in medicine: III. Temperance. PMID- 16704185 TI - President Marshall's message. PMID- 16704186 TI - Care of the congestive heart failure patient: the Care, Cure, and Core Model. PMID- 16704187 TI - When the leader has a group in a difficult situation. AB - A novice leader can be unnerved when faced with a group that has rigid time constraints and the personal responsibility to ensure group task is accomplished. The leader may become overwhelmed with obstacles that are difficult to control. However, use of careful group leadership strategies can make what seems like a daunting task doable and can allow the group to successfully meet goals and objectives. PMID- 16704188 TI - A word of caution and advice on study materials. PMID- 16704189 TI - Treatment of hypertension. PMID- 16704190 TI - Using the Giger-Davidhizar Transcultural Assessment Model (GDTAM) in providing patient care. PMID- 16704191 TI - Single-drug multiligand conjugates: synthesis and preliminary cytotoxicity evaluation of a paclitaxel-dipeptide "scorpion" molecule. AB - To improve the targeting properties of receptor-directed drug-peptide conjugates, a multiligand approach was proposed and a model "scorpion" conjugate (6, Figure 1), consisting of two peptide "claws" and a paclitaxel (PTX) "tail", was synthesized. The cell surface receptor-directed peptide used in this single-drug multiligand (SDML) model was a segment of the amphibian peptide bombesin (BBN) which had the Y6Q7W8A9V10G11H12L13M14-NH2 sequence, designated here as BBN[6-14] (2, Figure 2). Due to the lipophilic nature of both PTX and BBN[6-14], compound 6 had a low water solubility. To enhance the solubility, PEG derivatives of this conjugate were prepared with the polymer inserted either in the claws or in the tail regions. In a preliminary random screening, conjugate 6 showed superior cytotoxic activity in several GRPR-positive human cancer cell lines as compared to free PTX and two single-drug single-ligand (SDSL) conjugates. In a receptor blocking experiment, addition of excess unconjugated BBN[6-14] ligand reduced the cytotoxicity of conjugate 6, indicating the receptor-mediated mechanism of drug delivery. The PEG-derived conjugates showed activities which were intermediate between SDSL and the SDML congeners. Also, an increase in the number of the PEG segments lowered cytotoxicity, possibly due to steric hindrance against ligand receptor binding. Taken together, these results demonstrate the potential of the multiligand approach in the design of receptor-targeting conjugates for tumor specific drug delivery. PMID- 16704192 TI - Fast water-exchange Gd3+-(DO3A-like) complex functionalized with aza-15-crown-5 showing prolonged residence lifetime in vivo. AB - A bis-hydrated Gd3+ complex based on tris acetic acid-1,4,7,10 tetraazacyclododecane (DO3A) that was functionalized with aza-15-crown-5 demonstrated a nearly optimal water-exchanging rate (k(ex) = 3.1 x 10(7) s(-1)) and low acute cytotoxicity. Efficient magnetic resonance signal intensity enhancements and prolonged residence lifetime induced by this small molecular complex in vivo were demonstrated even with one-fifth of the standard dosage used in the clinic. PMID- 16704193 TI - Carbohydrate-labeled fluorescent microparticles and their binding to lectins. AB - The preparation of micrometer-sized, cross-linked poly(p-phenyleneethynylene) (PPE) beads to which simple monosaccharides are attached is reported. Mannose, glucose, and galactose derivatives have been synthesized. The fluorescence properties, size distribution, and morphology of these microparticles have been elucidated through fluorimetry, fluorescence confocal microscopy, and scanning electron microscopy. Protein binding assays were carried out using Concanavalin A tagged with the fluorophore Texas Red, and the resultant bioconjugates were imaged using confocal microscopy. The microparticles are shown to exhibit efficient binding to lectins and may have potential application as fluorescent probes, biocapture agents, or column packing material for affinity chromatography. PMID- 16704194 TI - Extension of the single amino acid chelate concept (SAAC) to bifunctional biotin analogues for complexation of the M(CO)3(+1) Core (M = Tc and Re): syntheses, characterization, biotinidase stability, and avidin binding. AB - Biotin and avidin form one of the most stable complexes known (K(D) = 10(-15) M( 1)) making this pairing attractive for a variety of biomedical applications including targeted radiotherapy. In this application, one of the pair is attached to a targeting molecule, while the other is subsequently used to deliver a radionuclide for imaging and/or therapeutic applications. Recently, we reported a new single amino acid chelate (SAAC) capable of forming stable complexes with Tc(CO)3 or Re(CO)3 cores. We describe here the application of SAAC analogues for the development of a series of novel radiolabeled biotin derivatives capable of forming robust complexes with both Tc and Re. Compounds were prepared through varying modification of the free carboxylic acid group of biotin. Each 99mTc complex of SAAC-biotin was studied for their ability to bind avidin, susceptibility to biotinidase, and specificity for avidin in an in vivo avidin containing tumor model. The radiochemical stability of the 99mTc(CO)3 complexes was also investigated by challenging each 99mTc-complex with large molar excesses of cysteine and histidine at elevated temperature. All compounds were radiochemically stable for greater than 24 h at elevated temperature in the presence of histidine and cysteine. Both [99mTc(CO)3(L6)]+1 [TcL6; L6 = biotinylamidopropyl-N,N-(dipicolyl)amine] and [99mTc(CO)3(L12a)]+1 (TcL12; L12 = N,N-(dipicolyl)biotinamido-Boc-lysine; TcL12a; L12a = N,N-(dipicolyl)biotinamide lysine) readily bound to avidin whereas [99mTc(CO)3(L9)]+1 [TcL9; L9 = N,N (dipicolyl)biotinamine] demonstrated minimal specific binding. TcL6 and TcL9 were resistant to biotinidase cleavage, while TcL12a, which contains a lysine linkage, was rapidly cleaved. The highest uptake in an in vivo avidin tumor model was exhibited by TcL6, followed by TcL9 and TcL12a, respectively. This is likely the result of both intact binding to avidin and resistance to circulating biotinidase. Ligand L6 is the first SAAC analogue of biotin to demonstrate potential as a radiolabeled targeting vector of biotin capable of forming robust radiochemical complexes with both 99mTc and rhenium radionuclides. Computational simulations were performed to assess biotin-derivative accommodation within the binding site of the avidin. These calculations predict that deformation of the surface domain of the binding pocket can occur to accommodate the transition metal-biotin derivatives with negligible changes to the inner-beta-barrel, the region most responsible for binding and retaining biotin and its derivatives. The biological activity and biodistribution of the technetium complexes TcL6, TcL9, and TcL12a were examined in an avidin tumor model. In the avidin bead tumor localization model, TcL6 demonstrated the most favorable localization with a 7:1 ratio of avidin bead implanted muscle versus normal muscle, while TcL9 exhibited a 2:1 ratio. However, TcL9 displayed no specificity for avidin. PMID- 16704195 TI - Isostructural Re and 99mTc complexes of biotin derivatives for fluorescence and radioimaging studies. AB - The reaction of biotinamine with two equivalents of 2-quinoline aldehyde in the presence of Na(OAc)3BH in dichloroethane provides N,N bis(methylquinoline)biotinamine (L1), a molecule displaying a tridentate donor terminus which has proven effective in coordinating to the {M(CO)3}+ core (M = Tc, Re). Reaction of L1 with (NEt4)2[Re(CO)3Br3] yields [Re(CO)3(L1)]Br, a compound with an absorbance at 350 nm and luminescence emission maxima at 425 and 580 nm. The luminescence lifetime of 11.4 mus, which is associated with the 580 nm emission, is sufficiently prolonged to enable time-gating techniques to be used during in vitro imaging studies and to overcome interference from endogenous fluorescence. Exposure of avidin beads to {Re(CO)3(L1)]Br resulted in binding, which was qualitatively imaged using fluorescence microscopy. The 99mTc analogue [99mTc(CO)3(L1)]+1 was prepared by reacting L1 with [99mTc(CO)3(H2O)3]+1 and purified by HPLC. The 99mTc complex is chemically robust and resistant to cysteine and histidine challenges. This study demonstrates that complementary fluorescent and radioactive biotin-derived probes may be readily prepared to allow direct correlation of in vitro and in vivo molecular imaging studies. PMID- 16704196 TI - Effects of cell-permeating peptide binding on the distribution of 125I-labeled Fab fragment in rats. AB - The peptides comprising the sequence of HIV-1 Tat protein (positions 48-60), Antennapedia (positions 43-58), and HIV-1 Rev protein (positions 34-50) are known to be cell-permeating. In this study, we examined how the distribution of Fab fragments in rats is affected by conjugation with these peptides. Fab fragment was iodinated by a chloramine-T method and then chemically conjugated with cell permeating peptide. The complex of 125I-Fab and cell-permeating peptide was administered to male rats intravenously at a dose of 1 mg/kg, and whole-body autoradiography was performed at 4 and 24 h after administration. The patterns of distribution of 125I-Fab exhibited remarkable variation depending on the cell permeating peptide used. In particular, at 4 h, high concentrations of radioactivity were observed in the spleen, adrenal gland, renal medulla, and liver with Rev peptide-Fab complex, in the liver and spleen with Tat peptide-Fab complex, and in the spleen, adrenal gland, and liver with Antennapedia peptide Fab complex. Even at 24 h, high concentrations of radioactivity were still observed in the spleen and renal medulla of rat with Rev peptide-Fab complex, and in the spleen and renal cortex of rat with Antennapedia peptide-Fab complex. These findings demonstrate that the patterns of distribution of peptide-125I-Fab complexes can be modulated by selection of cell-penetrating peptides. Moreover, the patterns of retention of peptide-125I-Fab complexes in internal organs also differed at 24 h after administration. These findings provide valuable information for the development of novel antibody pharmaceuticals and therapeutic systems. PMID- 16704197 TI - Synthesis and grafting of thioctic acid-PEG-folate conjugates onto Au nanoparticles for selective targeting of folate receptor-positive tumor cells. AB - This paper reports the creation of Au nanoparticles (AuNP) that are soluble in aqueous solution over a broad range of pH and ionic strength values and that are capable of selective uptake by folate receptor positive (FR+) cancer cells. A novel poly(ethylene glycol) (PEG) construct with thioctic acid and folic acid coupled on opposite ends of the polymer chain was synthesized for targeting the AuNP to FR+ tumor cells via receptor-mediated endocytosis. These folic acid-PEG thioctic acid conjugates were grafted onto 10-nm-diameter Au particles in aqueous solution. The resulting folate-PEG-coated nanoparticles do not aggregate over a pH range of from 2 to 12 and at electrolyte concentrations of up to 0.5 M NaCl with particle concentrations as high as 1.5 x 10(13) particles/mL. Transmission electron microscopy was used to document the performance of these coated nanoparticles in cell culture. Selective uptake of folate-PEG grafted AuNPs by KB cells, a FR+ cell line that overexpress the folate receptor, was observed. AuNP uptake was minimal in cells that (1) do not overexpress the folate receptor, (2) were exposed to AuNP lacking the folate-PEG conjugate, or (3) were co-incubated with free folic acid in large excess relative to the folate-PEG grafted AuNP. Understanding this process is an important step in the development of methods that use targeted metal nanoparticles for tumor imaging and ablation. PMID- 16704198 TI - Rapid production of functionalized recombinant proteins: marrying ligation independent cloning and in vitro protein ligation. AB - Functional genomics and proteomics have been very active fields since the sequencing of several genomes was completed. To assign a physiological role to the newly discovered coding genes with unknown function, new generic methods for protein production, purification, and targeted functionalization are needed. This work presents a new vector, pCYSLIC, that allows rapid generation of Escherichia coli expression constructs via ligation-independent cloning (LIC). The vector is designed to facilitate protein purification by either Ni-NTA or GSH affinity chromatography. Subsequent proteolytic removal of affinity tags liberates an N terminal cysteine residue that is then used for covalent modification of the target protein with different biophysical probes via protein ligation. The described system has been tested on 36 mammalian Rab GTPases, and it was demonstrated that recombinant GTPases produced with pCYSLIC could be efficiently modified with fluorescein or biotin in vitro. Finally, LIC was compared with the recently developed In-Fusion cloning method, and it was demonstrated that In Fusion provides superior flexibility in choice of expression vector. By the application of In-Fusion cloning Cys-Rab6A GTPase with an N-terminal cysteine residue was generated employing unmodified pET30a vector and TVMV protease. PMID- 16704199 TI - Structure-function engineering of interferon-beta-1b for improving stability, solubility, potency, immunogenicity, and pharmacokinetic properties by site selective mono-PEGylation. AB - Recombinant interferon-beta-1b (IFN-beta-1b) is used clinically in the treatment of multiple sclerosis. In common with many biological ligands, IFN-beta-1b exhibits a relatively short serum half-life, and bioavailability may be further diminished by neutralizing antibodies. While PEGylation is an approach commonly employed to increase the blood residency time of protein therapeutics, there is a further requisite for molecular engineering approaches to also address the stability, solubility, aggregation, immunogenicity and in vivo exposure of therapeutic proteins. We investigated these five parameters of recombinant human IFN-beta-1b in over 20 site-selective mono-PEGylated or multi-PEGylated IFN-beta 1b bioconjugates. Primary amines were modified by single or multiple attachments of poly(ethylene glycol), either site-specifically at the N-terminus, or randomly on the 11 lysines. In two alternate approaches, site-directed mutagenesis was independently employed in the construction of designed IFN-beta-1b variants containing either a single free cysteine or lysine for site-specific PEGylation. Optimization of conjugate preparation with 12 kDa, 20 kDa, 30 kDa, and 40 kDa amine-selective PEG polymers was achieved, and a comparison of the structural and functional properties of the IFN-beta-1b proteins and their PEGylated counterparts was conducted. Peptide mapping and MALDI-TOF mass spectrometric analysis confirmed the attachment sites of the PEG polymer. Independent biochemical and bioactivity analyses, including antiviral and antiproliferation bioassays, circular dichroism, capillary electrophoresis, flow cytometric profiling, reversed phase and size exclusion HPLC, and immunoassays demonstrated that the functional activities of the designed IFN-beta-1b conjugates were maintained, while the formation of soluble or insoluble aggregates of IFN-beta-1b was ameliorated. Immunogenicity and pharmacokinetic studies of selected PEGylated IFN-beta-1b compounds in mice and rats demonstrated both diminished IgG responses, and over 100-fold expanded AUC exposure relative to the unmodified protein. The results demonstrate the capacity of this macromolecular engineering strategy to address both pharmacological and formulation challenges for a highly hydrophobic, aggregation-prone protein. The properties of a lead mono-PEGylated candidate, 40 kDa PEG2-IFN-beta-1b, were further investigated in formulation optimization and biological studies. PMID- 16704200 TI - A molecular toolkit for highly insulating tethered bilayer lipid membranes on various substrates. AB - Tethered bilayer lipid membranes (tBLMs) are promising model architectures that mimic the structure and function of natural biomembranes. They provide a fluid, stable, and electrically sealing platform for the study of membrane related processes, specifically, the function of incorporated membrane proteins. This paper presents a generic approach toward the synthesis of functional tBLMs adapted for application to various surfaces. The central element of a tethered membrane consists of a lipid bilayer. Its proximal layer is covalently attached via a spacer unit to a solid support, either gold or silicon oxide. The membranes are characterized optically by using surface plasmon resonance spectroscopy (SPR) or ellipsometry and electrically by using electrochemical impedance spectroscopy (EIS). The bilayer membranes obtained show high electrical barrier properties and can be used to incorporate and study small membrane proteins in a functional form. PMID- 16704201 TI - Bioconjugatable porphyrins bearing a compact swallowtail motif for water solubility. AB - A broad range of applications requires access to water-soluble, bioconjugatable porphyrins. Branched alkyl groups attached at the branching site to the porphyrin meso position are known to impart high organic solubility. Such "swallowtail" motifs bearing a polar group (hydroxy, dihydroxyphosphoryl, dihydroxyphosphoryloxy) at the terminus of each branch have now been incorporated at a meso site in trans-AB-porphyrins. The incorporation of the swallowtail motif relies on rational synthetic methods whereby a 1,9-bis(N propylimino)dipyrromethane (bearing a bioconjugatable tether at the 5-position) is condensed with a dipyrromethane (bearing a protected 1,5-dihydroxypent-3-yl unit at the 5-position). The two hydroxy groups in the swallowtail motif of each of the resulting zinc porphyrins can be transformed to the corresponding diphosphate or diphosphonate product. A 4-(carboxymethyloxy)phenyl group provides the bioconjugatable tether. The six such porphyrins reported here are highly water-soluble (> or =20 mM at room temperature in water at pH 7) as determined by visual inspection, UV-vis absorption spectroscopy, or 1H NMR spectroscopy. Covalent attachment was carried out in aqueous solution with the unprotected porphyrin diphosphonate and a monoclonal antibody against the T-cell receptor CD3epsilon. The resulting conjugate performed comparably to a commercially available fluorescein isothiocyanate-labeled antibody with Jurkat cells in flow cytometry and fluorescence microscopy assays. Taken together, this work enables preparation of useful quantities of water-soluble, bioconjugatable porphyrins in a compact architecture for applications in the life sciences. PMID- 16704202 TI - Fractionation of serum components using nanoporous substrates. AB - Numerous previously uncharacterized molecules resident within the low molecular weight circulatory proteome may provide a picture of the ongoing pathophysiology of an organism. Recently, proteomic signatures composed of low molecular weight molecules have been identified using mass spectrometry combined with bioinformatic algorithms. Attempts to sequence and identify the molecules that underpin the fingerprints are currently underway. The finding that many of these low molecular weight molecules may exist bound to circulating carrier proteins affords a new opportunity for fractionation and separation techniques prior to mass spectrometry-based analysis. In this study we demonstrate a method whereby nanoporous substrates may be used for the facile and reproducible fractionation and selective binding of the serum-based biomarker material, including subcellular proteins found within the serum. Aminopropyl-coated nanoporous silicon, when exposed to serum, can deplete serum of proteins and yield a serum with a distinct, altered MS profile. Additionally, aminopropyl-coated, nanoporous controlled-pore glass beads are able to bind a subset of serum proteins and release them with stringent elution. The eluted proteins have distinct MS profiles, gel electrophoresis profiles, and differential peptide sequence identities, which vary based on the size of the nanopores. These material surfaces could be employed in strategies for the harvesting and preservation of labile and carrier-protein-bound molecules in the blood. PMID- 16704203 TI - Near-infrared fluorescent deoxyglucose analogue for tumor optical imaging in cell culture and living mice. AB - 2-deoxy-2-[18F]fluoro-D-glucose ([18F]FDG) has extensively been used for clinical diagnosis, staging, and therapy monitoring of cancer and other diseases. Nonradioactive glucose analogues enabling the screening of the glucose metabolic rate of tumors are of particular interest for anticancer drug development. A nonradioactive fluorescent deoxyglucose analogue may have many applications for both imaging of tumors and monitoring therapeutic efficacy of drugs in living animals and may eventually translate to clinical applications. We found that a fluorescent 2-deoxyglucose analogue, 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4 yl)amino]-2-deoxy-D-glucose (2-NBDG), can be delivered in several tumor cells via the glucose transporters (GLUTs). We therefore conjugated D-glucosamine with a near-infrared (NIR) fluorphor Cy5.5 and tested the feasibility of the Cy5.5-D glucosamine (Cy5.5-2DG) conjugate for NIR fluorescence imaging of tumors in a preclinical xenograft animal model. Cy5.5-2DG was prepared by conjugating Cy5.5 monofunctional N-hydroxysuccinimide ester (Cy5.5-NHS) and D-glucosamine followed by high-performance liquid chromatography purification. The accumulation of Cy5.5 2DG and Cy5.5-NHS in different tumor cell lines at 37 and 4 degrees C were imaged using a fluorescence microscope. Tumor targeting and retention of Cy5.5-2DG and Cy5.5-NHS in a subcutaneous U87MG glioma and A375M melanoma tumor model were evaluated and quantified by a Xenogen IVIS 200 optical cooled charged-coupled device system. Fluorescence microscopy imaging shows that Cy5.5-2DG and Cy5.5-NHS are taken up and trapped by a variety of tumor cell lines at 37 degrees C incubation, while they exhibit marginal uptake at 4 degrees C. The tumor cell uptake of Cy5.5-2DG cannot be blocked by the 50 mM D-glucose, suggesting that Cy5.5-2DG may not be delivered in tumor cells by GLUTs. U87MG and A375M tumor localization was clearly visualized in living mice with both NIR fluorescent probes. Tumor/muscle contrast was clearly visible as early as 30 min postinjection (pi), and the highest U87MG tumor/muscle ratios of 2.81 +/- 0.10 and 3.34 +/- 0.23 were achieved 24 h pi for Cy5.5-2DG and Cy5.5-NHS, respectively. While as a comparison, the micropositron emission tomography imaging study shows that [18F]FDG preferentially localizes to the U87MG tumor, with resulting tumor/muscle ratios ranging from 3.89 to 4.08 after 30 min to 2 h postadministration of the probe. In conclusion, the NIR fluorescent glucose analogues, Cy5.5-2DG and Cy5.5-NHS, both demonstrate tumor-targeting abilities in cell culture and living mice. More studies are warranted to further explore their application for optical tumor imaging. To develop NIR glucose analogues with the ability to target GLUTs/hexokinase, it is highly important to select NIR dyes with a reasonable molecular size. PMID- 16704204 TI - A blue-green absorbing cross-linker for rapid photoswitching of peptide helix content. AB - Azobenzene derivatives can be used to reversibly photoregulate secondary structure when introduced as intramolecular bridges in peptides and proteins. Here we report the design, synthesis, and characterization of a disubstituted N,N dialkyl azobenzene derivative that absorbs near 480 nm in aqueous solution and relaxes with a half-life of approximately 50 ms at room temperature. The wavelength of maximum absorbance and the rate of thermal relaxation are solvent dependent. An increase in the percentage of organic solvent leads, in general, to a blue shift in the absorbance maximum and a slowing of the relaxation rate. In accordance with the design, the thermal relaxation of the azobenzene cross-linker from cis to trans causes an increase in the helix content of one peptide where the linker is attached via cysteine residues spaced at i, i + 11 positions and a decrease in helix content of another peptide with cysteine residues spaced at i, i + 7. This cross-linker design thus expands the possibilities for fast photocontrol of peptide and protein structure. PMID- 16704205 TI - pH-responsive three-layered PEGylated polyplex micelle based on a lactosylated ABC triblock copolymer as a targetable and endosome-disruptive nonviral gene vector. AB - Nonviral vectors for gene therapy have recently received an increased impetus because of the inherent safety problems of the viral vectors, while their transfection efficiency is generally low compared to the viral vectors. The lack of the ability to escape from the endosomal compartments is believed to be one of the critical barriers to the intracellular delivery of noviral gene vectors. This study was devoted to the design and preparation of a novel ABC triblock copolymer for constructing a pH-responsive and targetable nonviral gene vector. The copolymer, lactosylated poly(ethylene glycol)-block-poly(silamine)-block-poly[2 (N,N-dimethylamino)ethyl methacrylate] (Lac-PEG-PSAO-PAMA), consists of lactosylated poly(ethylene glycol) (A-segment), a pH-responsive polyamine segment (B-segment), and a DNA-condensing polyamine segment (C-segment). The Lac-PEG-PSAO PAMA spontaneously associated with plasmid DNA (pDNA) to form three-layered polyplex micelles with a PAMA/pDNA polyion complex (PIC) core, an uncomplexed PSAO inner shell, and a lactosylated PEG outer shell, as confirmed by 1H NMR spectroscopy. Under physiological conditions, the Lac-PEG-PSAO-PAMA/pDNA polyplex micelles prepared at an N/P (number of amino groups in the copolymer/number of phosphate groups in pDNA) ratio above 3 were found to be able to condense pDNA, thus adopting a relatively small size (< 150 nm) and an almost neutral surface charge (zeta approximately +5 mV). The micelle underwent a pH-induced size variation (pH = 7.4, 132.6 nm --> pH = 4.0, 181.8 nm) presumably due to the conformational changes (globule-rod transition) of the uncomplexed PSAO chain in response to pH, leading to swelling of the free PSAO inner shell at lowered pH while retaining the condensed pDNA in the PAMA/pDNA PIC core. Furthermore, the micelles exhibited a specific cellular uptake into HuH-7 cells (hepatocytes) through asialoglycoprotein (ASGP) receptor-mediated endocytosis and achieved a far more efficient transfection ability of a reporter gene compared to the Lac PEG-PSAO/pDNA and Lac-PEG-PAMA/pDNA polyplex micelles composed of the diblock copolymers and pDNA. The effect of hydroxychloroquine as an endosomolytic agent on the transfection efficiency was not observed for the Lac-PEG-PSAO-PAMA/pDNA polyplex micelles, whereas the nigericin treatment of the cell as an inhibitor for the endosomal acidification induced a substantial decrease in the transfection efficiency, suggesting that the protonation of the free PSAO inner shell in response to a pH decrease in the endosome might lead to the disruption of the endosome through buffering of the endosomal cavity. Therefore, the polyplex micelle composed of ABC (ligand-PEG/pH-responsive segment/DNA-condensing segment) triblock copolymer would be a promising approach to a targetable and endosome disruptive nonviral gene vector. PMID- 16704207 TI - Diethylenetriaminepentaacetic acid-gadolinium (DTPA-Gd)-conjugated polysuccinimide derivatives as magnetic resonance imaging contrast agents. AB - Biocompatible polysuccinimide (PSI) derivatives conjugated with diethylenetriaminepentaacetic acid gadolinium (DTPA-Gd) were prepared as magnetic resonance imaging (MRI) contrast agents. In this study, we synthesized PSI derivatives incorporating methoxy-poly(ethylene glycol) (mPEG) as hydrophilic ligand, hexadecylamine as hydrophobic ligand, and DTPA-Gd as contrast agent. PSI was synthesized by the polycondensation polymerization of aspartic acid. All the synthesized materials were characterized by proton nuclear magnetic resonance (1H NMR). Critical micellization concentrations were determined using fluorescent probes (pyrene). Micelle size and shape were measured by electro-photometer light scattering (ELS) and atomic force microscopy (AFM). The formed micelle size ranged from 100 to 300 nm. The T1-weighted MR images of the phantom prepared with PSI-mPEG-C16-(DTPA-Gd) were obtained in a 3.0 T clinical MR imager, and the conjugates showed a great potential as MRI contrast agents. PMID- 16704206 TI - Synthesis of a biotin derivative of iberiotoxin: binding interactions with streptavidin and the BK Ca2+-activated K+ channel expressed in a human cell line. AB - Iberiotoxin (IbTx) is a scorpion venom peptide that inhibits BK Ca2+-activated K+ channels with high affinity and specificity. Automated solid-phase synthesis was used to prepare a biotin-labeled derivative (IbTx-LC-biotin) of IbTx by substitution of Asp19 of the native 37-residue peptide with N--(D-biotin-6 amidocaproate)-L-lysine. Both IbTx-LC-biotin and its complex with streptavidin (StrAv) block single BK channels from rat skeletal muscle with nanomolar affinity, indicating that the biotin-labeled residue, either alone or in complex with StrAv, does not obstruct the toxin binding interaction with the BK channel. IbTx-LC-biotin exhibits high affinity (KD = 26 nM) and a slow dissociation rate (koff = 5.4 x 10(-4) s(-1)) in a macroscopic blocking assay of whole-cell current of the cloned human BK channel. Titration of IbTx-LC-biotin with StrAv monitored by high performance size exclusion chromatography is consistent with a stoichiometry of two binding sites for IbTx-LC-biotin per StrAv tetramer, indicating that steric interference hinders simultaneous binding of two toxin molecules on each of the two biotin-binding faces of StrAv. In combination with fluorescent conjugates of StrAv or anti-biotin antibody, IbTx-LC-biotin was used to image the surface distribution of BK channels on a transfected cell line. Fluorescence microscopy revealed a patch-like surface distribution of BK channel protein. The results support the feasibility of using IbTx-LC-biotin and similar biotin-tagged K+ channel toxins for diverse applications in cellular neurobiology. . PMID- 16704208 TI - Combination cancer therapy by hapten-targeted prodrug-activating enzymes and cytokines. AB - Combination therapy can help overcome limitations in the treatment of heterogeneous tumors. In the current study, we examined whether multiple therapeutic agents could be targeted to anti-dansyl single-chain antibodies (DNS scFv) that were anchored on the plasma membrane of cancer cells. Functional DNS scFv could be stably expressed on CT-26 colon cancer cells both in vitro and in vivo. Dansyl moieties were covalently attached to recombinant beta-glucuronidase (betaG) and interleukin 2 (IL-2) via a flexible poly(ethylene glycol) linker to form DNS-PEG-betaG and DNS-PEG-IL-2 conjugates. The conjugates displayed enzymatic and splenocyte-stimulatory activities, respectively, that were similar to those of the unmodified proteins. The conjugates selectively bound CT-26 cells that expressed anti-DNS scFv (CT-26/DNS cells) but not CT-26 cells that expressed control scFv (CT-26/phOx cells). DNS-PEG-betaG preferentially activated a glucuronide prodrug (BHAMG) of p-hydroxy aniline mustard at CT-26/DNS cells in culture and accumulated in subcutaneous CT-26/DNS tumors after intravenous administration. Systemic administration of DNS-PEG-IL-2 or DNS-PEG-betaG and BHAMG significantly delayed the growth of CT-26/DNS but not control CT-26/phOx tumors. Combination treatment with DNS-PEG-betaG and BHAMG followed by DNS-PEG-IL 2 therapy significantly suppressed the growth of CT-26/DNS tumors as compared to either single-agent regimen. These results show that at least two DNS-modified therapeutic agents can be selectively delivered to DNS scFv receptors in vitro and in vivo, allowing combination therapy of DNS scFv-modified tumors. PMID- 16704209 TI - Efficient DNA alkylation by a pyrrole-imidazole CBI conjugate with an indole linker: sequence-specific alkylation with nine-base-pair recognition. AB - Conjugates 7, 8, and 10 of N-methylpyrrole (Py)-N-methylimidazole (Im) polyamides and 1,2,9,9a-tetrahydrocyclopropa[1,2-c]benz[1,2-e]indol-4-one (CBI) with a 5 amino-1H-indole-2-carbonyl linker were synthesized by Fmoc solid-phase synthesis and a subsequent liquid-phase coupling procedure. The DNA alkylating abilities of conjugates 7, 8, 6b, and 10 were examined using Texas Red-labeled PCR fragments and high-resolution denaturing gel electrophoresis. CBI conjugates 7 and 8 exhibited highly efficient sequence-specific DNA alkylation comparable with previous CBI conjugates with a vinyl linker. In particular, conjugate 10, with a 10-ringed hairpin Py-Im polyamide, alkylated at the adenine of 5'-ACAAATCCA-3'. Introduction of an indole linker greatly facilitated the synthesis of sequence specific alkylating Py-Im polyamides. PMID- 16704210 TI - Folate immobilized and PEGylated adenovirus for retargeting to tumor cells. AB - The surface of adenovirus (ADV) was modified with folate-poly(ethylene glycol) (FOL-PEG) conjugate to overcome the inherent problems associated with viral gene therapy. The ADV coding the green fluorescent protein (GFP) gene was used to immobilize PEG or FOL-PEG conjugate on the viral surface to comparatively evaluate their extents of retargeting and immune response, as compared to those of naked ADV. The FOL-PEG immobilized ADV exhibited a greatly enhanced level of GFP expression than naked or PEG immobilized ADV for a folate receptor overexpressing cell line (KB cells), but not for a folate receptor deficient cell line (A549 cells), suggesting that the retargeting of ADV could be achieved by immobilizing FOL-PEG conjugate. The ADV immobilized with PEG or FOL-PEG also significantly lowered innate immune response, as judged from the amount of interleukin 6 released from macrophage cells. PMID- 16704211 TI - Interaction of polycationic polymers with supported lipid bilayers and cells: nanoscale hole formation and enhanced membrane permeability. AB - Interactions of polycationic polymers with supported 1,2-dimyristoyl-sn-glycero-3 phosphocholine (DMPC) lipid bilayers and live cell membranes (KB and Rat2) have been investigated using atomic force microscopy (AFM), cytosolic enzyme assays, confocal laser scanning microscopy (CLSM), and a fluorescence-activated cell sorter (FACS). Polycationic polymers poly-L-lysine (PLL), polyethylenimine (PEI), and diethylaminoethyl-dextran (DEAE-DEX) and sphere-like poly(amidoamine) (PAMAM) dendrimers are employed because of their importance for gene and drug delivery. AFM studies indicate that all the polycationic polymers cause the formation and/or expansion of preexisting defects in supported DMPC bilayers in the concentration range of 1-3 microg/mL. By way of contrast, hydroxyl-containing neutral linear poly(ethylene glycol) (PEG) and poly(vinyl alcohol) (PVA) do not induce hole formation or expand the size of preexisting defects in the same concentration range. All polymers tested are not toxic to KB or Rat2 cells up to a 12 microg/mL concentration (XTT assay). In the concentration range of 6-12 microg/mL, however, significant amounts of the cytosolic enzymes lactate dehydrogenase (LDH) and luciferase (LUC) are released. PEI, which possesses the greatest density of charged groups on its chain, shows the most dramatic increase in membrane permeability. In addition, treatment with polycationic polymers allows the small dye molecules propidium idodide (PI) and fluorescein (FITC) to diffuse in and out of the cells. CLSM images also show internalization of PLL labeled with FITC dye. In contrast, controls of membrane permeability using the neutral linear polymers PEG and PVA show dramatically less LDH and LUC leakage and no enhanced dye diffusion. Taken together, these data are consistent with the hypothesis that polycationic polymers induce the formation of transient, nanoscale holes in living cells and that these holes allow a greatly enhanced exchange of materials across the cell membrane. PMID- 16704212 TI - A peripheral benzodiazepine receptor targeted agent for in vitro imaging and screening. AB - We developed a molecular imaging agent (MIA), a conjugable form of PK11195 (conPK11195) coupled to a lissamine dye (Liss-ConPK11195), which targets the peripheral benzodiazepine receptor (PBR). To determine that our compound specifically binds to this 18 kDa protein, primarily expressed on the mitochondria, we performed classic binding studies on live MDA-MB-231 breast cancer cells and measured fluorescence in cell fractions of C6 glioma cells. We found that conPK11195 conjugated to the fluorophore retained significant binding to its target. Here we demonstrate the utility of the agent for in vitro imaging of live cells by specific binding to the protein of interest. PMID- 16704213 TI - Annexin A5-functionalized bimodal lipid-based contrast agents for the detection of apoptosis. AB - Apoptosis, or programmed cell death, plays an important role in the etiology of a variety of diseases, including cancer and myocardial infarction. Visualization of apoptosis would allow both early detection of therapy efficiency and evaluation of disease progression. To that aim, we synthesized two types of lipid-based bimodal contrast agents that enable the detection of apoptotic cells with both MRI and optical techniques. MR contrast was provided either by entrapment of iron oxide particles within pegylated micelles or by incorporation of Gd-DTPA bis(stearylamide) (Gd-DTPA-BSA) lipids within the lipid bilayer of pegylated liposomes. The resulting contrast agents were approximately 10 and 100 nm in diameter, respectively. Additional fluorescent lipids were incorporated in the lipid (bi)layer of the contrast agents to allow parallel detection with optical methods. Multiple human recombinant annexin A5 molecules were covalently coupled to introduce specificity for apoptotic cells. Both annexin A5-conjugated contrast agents were shown to significantly increase the relaxation rates of apoptotic cell pellets compared to untreated control cells and apoptotic cells that were treated with nonfunctionalized nanoparticles. Increased relaxation rates were confirmed to originate from association of the contrast agents to apoptotic cells by confocal microscopy. The targeted nanoparticles presented in this study, which differ both in size and in magnetic properties, may have applications for the in vivo detection of apoptosis. PMID- 16704214 TI - Evaluation of cell-penetrating peptides (CPPs) as vehicles for intracellular delivery of antisense peptide nucleic acid (PNA). AB - Cell-penetrating peptides (CPPs) are characterized by their ability to be internalized in mammalian cells. To investigate the relative potency of CPPs as carriers of medicinally relevant cargo, a positive read-out assay based on the ability of a peptide nucleic acid (PNA) oligomer to promote correct expression of a recombinant luciferase gene was employed. Seven different CPPs were included in the study: Transportan, oligo-arginine (R7-9), pTat, Penetratin, KFF, SynB3, and NLS. The CPP-PNA conjugates were synthesized by different conjugation chemistries: continuous synthesis, maleimide coupling, and ester or disulfide linkage. Under serum-free conditions PNA-SS-Transportan-amide (ortho)-PNA was found to be the most potent conjugate, resulting in maximum luciferase signal at a concentration of 1-2 microM. (D-Arg)9-PNA showed optimal efficacy at 5 microM but gave rise to only one-third of the luciferase signal obtained with the Transportan conjugate. The pTat- and KFF-PNA conjugates showed significantly lower efficacy. The penetratin-, SynB3-. and NLS-PNA conjugates showed only minimal or no activity. Serum was found to have a drastic negative impact on CPP driven cellular uptake. PNA-SS-Transportan-acid (ortho) and (D-Arg)9-PNA were least sensitive to the presence of serum. Both the chemical nature and, in the case of Transportan, the position of the peptide PNA coupling were found to have a major impact on the transport capacity of the peptides. However, no simple relationship between linker type and antisense activity of the conjugates could be deduced from the data. PMID- 16704215 TI - Linear topology confers in vivo gene transfer activity to polyethylenimines. AB - Although polyethylenimines (PEIs) are frequently used transfection agents, it is still unclear which of their properties are required for efficient gene delivery. This is even more striking when working in vivo since some PEIs are able to generate significant gene expression, whereas others are not. To facilitate a rational development of compounds with improved transfection activities, studies aimed at identifying the properties involved in the transfection process seem indispensable. In the present work, we investigated how transfection with linear PEI of 22 kDa allows for high reporter gene expression in lungs after intravenous injection, whereas the branched PEI of 25 kDa does not. To this end, we synthesized L-PEI derivatives that are intermediates between linear and branched PEIs. Our results show that the topology plays a crucial role in obtaining in vivo reporter gene expression, whereas the content of primary, secondary, and tertiary amines is only of minor importance. PMID- 16704216 TI - Temperature dependent gene expression induced by PNIPAM-based copolymers: potential of hyperthermia in gene transfer. AB - The objective of this work was to obtain gene delivery vectors with high efficiency induced by application of local hyperthermia. As a building construct for the polyplex particles, block copolymers were used, in which one block represents poly(ethyleneimine) (PEI) and another block a statistical copolymer of poly(N-isopropylacryamide) (PNIPAM) and different hydrophilic monomers (acrylamide or vinylpyrrolidinone). The block copolymers were synthesizized by radical polymerization of the corresponding monomers directly onto PEI. The complexation of DNA with these copolymers led to small, charge neutral particles, which aggregated upon increasing the temperature from 37 degrees C to 42 degrees C. This aggregation was found to be responsible for the enhanced transfection efficiency of these formulations under hyperthermic conditions. Gene expression in cells treated by hyperthermia was found to be nearly 2 orders of magnitude higher in comparison to cells transfected at physiological temperature. The mechanism by which hyperthermia influences the gene transfection efficiency is proposed. PMID- 16704217 TI - A new class of Gd-based DO3A-ethylamine-derived targeted contrast agents for MR and optical imaging. AB - Synthetic bifunctional probes based on [4,7-bis-carboxymethyl-10-(2-aminoethyl) 1,4,7,10-tetraaza-cyclododec-1-yl]-acetic acid (DO3A-ethylamine) preloaded with gadolinium were prepared for applications in targeted magnetic resonance imaging (MRI) and optical imaging. A convenient route of synthesis is reported, which allowed conjugation of this probe with biomolecules for the preparation of model MR contrast agents for targeted imaging. The conjugated probes have the following interesting properties: GdDO3A-ethylamido-biotin (Gd-9) can be used for targeted imaging using an avidin-biotin system. The fluorescent probe GdDO3A-ethylthiourea fluorescein (Gd-12) is a bimodal compound, which can be used for both MR and optical imaging. The precursors, DO3A-ethylamidopropyl-maleimide and DO3A-ethyl isothiocyanate contain a highly reactive moiety, which can interact with free SH terminals and N-terminals of biological molecules, respectively. In vitro MR relaxivity studies were performed at 300 MHz using different concentrations and chemical environments. MR relaxivity for ligand Gd-9 at pH 7.4, r1 was (3.32 +/- 0.03) s(-1) mM(-1) and r2 was (5.02 +/- 0.14) s(-1) mM(-1). For the mixture of Gd 9 with avidin, at pH 7.4, relaxivity increased linearly with the avidin concentration. A relaxivity enhancement of 45% for r1 and more than 400% for r2 with respect to the unbound biotinylated Gd3+ complex was found at a ratio of 4:1. MR relaxivity for ligand Gd-12, r1 was (5.36 +/- 0.05) s(-1) mM(-1) at pH 7.4. Fluorescence microscopy and spectroscopy of Gd-12-labeled 3T3 mouse fibroblasts showed a concentration-dependent intracellular uptake, accompanied by a slight dose-dependent increase in toxicity up to 150 microM. MR studies on labeled cells indicated a contrast enhancement in both T1- and T2-weighted images by the internalized compound, with the effect being more pronounced in T2 weighted images. Our results indicate that DO3A-ethylamine is a multipurpose precursor, from which various targeted contrast agents can be synthesized after a single-step conjugation with organic/bioorganic molecules. PMID- 16704218 TI - Nonviral gene carriers based on diblock copolymers of poly(2-ethyl-2-oxazoline) and linear polyethylenimine. AB - Diblock copolymers that consist of poly(2-ethyl-2-oxazoline) (PEOz) and linear polyethylenimine (LPEI) were prepared for use as nonviral gene carriers. The PEOz b-LPEI copolymers were synthesized by coupling PEOz with LPEI in a thiol disulfide exchange reaction between the sulfhydryl and pyridyl disulfide terminal groups. A polymer/DNA weight ratio (P/D) of over 12 was required to enable PEOz-b LPEI to condense DNA completely. The DNA-condensing capability of the diblock copolymers was increased with increasing the hydrolytic degrees of the LPEI segment. The PEOz-b-LPEI polyplexes were stable in 150 mM NaCl aqueous solution and had a mean diameter around 190 nm, whereas BPEI and LPEI polyplexes formed large aggregates in the range 300-500 nm. In addition, these polyplexes exhibited the sensitivity to solution pH and were dissociated in the acidic buffers (pH < or = 5.5). The results of in vitro cell viability and luciferase assay indicated that PEOz-b-LPEI showed not only low cytotoxicity but also high transfection efficiency in gene expression. PMID- 16704219 TI - Intracellular cargo delivery by an octaarginine transporter adapted to target prostate cancer cells through cell surface protease activation. AB - Delivery of therapeutics and imaging agents to target tissues requires localization and activation strategies with molecular specificity. Cell associated proteases can be used for these purposes in a number of pathologic conditions, and their enzymatic activities can be exploited for activation strategies. Here, molecules based on the d-arginine octamer (r8) protein transduction domain (PTD, also referred to as molecular transporters) have been adapted for selective uptake into cells only after proteolytic cleavage of a PTD attenuating sequence by the prostate-specific antigen (PSA), an extracellular protease associated with the surface and microenvironment of certain prostate cancer cells. Convergent syntheses of these activatable PTDs (APTDs) are described, and the most effective r8 PTD-attenuating sequence is identified. The conjugates are shown to be stable in serum, cleaved by PSA, and taken up into Jurkat (human T cells) and PC3M prostate cancer cell lines only after cleavage by PSA. These APTD peptide-based molecules may facilitate targeted delivery of therapeutics or imaging agents to PSA-expressing prostate cancers. PMID- 16704220 TI - Synthesis and in vitro/in vivo evaluation of novel 99mTc(CO)3-folates. AB - Novel organometallic 99mTc(I)-folate derivatives have been synthesized and evaluated in vitro and in vivo in order to assess the influence of the overall charge of the radioconjugates and the spacer entity on the affinity and pharmacokinetic profile. Folic acid has been functionalized at the gamma carboxylate group of the glutamate moiety with (i) a hydrophilic diethoxyethyl spacer bearing a picolylamine monoacetic acid chelate, (ii) a hexyl spacer bearing an iminodiacetic acid chelate, and (iii) a hexyl spacer with a bis(pyridylmethyl)amine chelating system. Coordination of the 99mTc(CO)3-core resulted in neutral complex 21, anionic complex 22, and cationic complex 23 in excellent yields (>90%) at ligand concentrations of 10(-4) M. Complexes 21-23 were HPLC purified for in vitro and in vivo experiments. In the case of 23, separation from the unlabeled folate analogue was incomplete, leading to low specific activity and, hence, significantly inferior in vivo uptake in folate receptor-positive (FR-positive) organs and tissues (tumors and kidneys). Time dependent in vivo studies were performed in female, athymic nude mice bearing subcutaneous FR-positive human KB cell xenografts at 1, 4, and 24 h post injection (p.i.) of the radiotracers. Tumor uptake ranged between 1.9-2.7% ID/g, 4 h p.i. and 1.6-2.2% ID/g, 24 h p.i. for 21 and 22, and 0.9% ID/g, 4 h p.i. and 1.1% ID/g, 24 h p.i. for 23. Blood clearance was fast for all derivatives (< or =0.2% ID/g 1 h p.i.). Significant fractions of radioactivity were found in nontargeted and FR-negative organs and tissues (particularly in the liver and the intestines/intestinal contents) at early time points p.i. Coadministration of folic acid reduced radioactivity in FR-positive tissues and organs to background levels. In conclusion, overall charge and the nature of the spacer entity seemed to have a relatively minor influence on receptor affinity and the in vivo pharmacokinetic profile of the tested radiofolates. PMID- 16704221 TI - Cyclic and acyclic oxorhenium(V)-peptide conjugates as new ligands of the human cyclophilin hCyp-18. AB - Peptide metalloconstructs display interesting conformations, activities, and resistance to proteolysis. However, introduction of a metal core close to the residues that interact with the protein might strongly affect the binding. We investigated the effects of a coordinated oxorhenium core on the binding of model peptides to cyclophilin hCyp-18, a protein implicated in important biological processes and several diseases. For this purpose, we synthesized a series of linear metalloconstructs bearing an oxorhenium(V) core (ReO3+), as well as a peptide cyclized through oxorhenium(V) coordination. All these peptides contain an Ala-Pro-Xaa-pNA moiety (Xaa = Cys derivative) and are anticipated to bind simultaneously to the S1-S1' and S2'-S3' subsites of hCyp-18. Therefore, the metal core is coordinated to both the cysteine residue and exogenous or endogenous NS2 tridentate systems. Cyclization of the peptide through metal coordination did not affect the affinity whereas bimolecular oxorhenium metalloconstructs bind hCyp-18 with a slightly better affinity than the corresponding nonmetalated peptide. Peptide labeling with a 99mTcO3+ core was also carried out successfully. PMID- 16704222 TI - Identification of formaldehyde-induced modifications in proteins: reactions with insulin. AB - Formaldehyde is frequently used to inactivate, stabilize, or immobilize proteins. The treatment results in a large variety of chemical modifications in proteins, such as the formation of methylol groups, Schiff bases, and methylene bridges. The purpose of the present study was to identify the stable formaldehyde-induced modifications in a small protein. Therefore, insulin was treated with excess formaldehyde (CH2O) or deuterated formaldehyde (CD2O). In a separate experiment, insulin was modified by formaldehyde (CH2O vs CD2O) and glycine. The mixture of CH2O-treated and CD2O-treated insulin was digested by the proteinase Glu-C. The peptide fragments obtained were analyzed by liquid chromatography-mass spectrometry (LC-MS). Seven intramolecular cross-links were identified in formaldehyde-treated insulin. Furthermore, eight out of the sixteen potentially reactive sites of the insulin molecule were modified by incubation with formaldehyde and glycine. Both the location and the chemical nature of the modifications could be assigned based on the mass increase of potential adducts as elucidated in our previous study (B. Metz et al. (2004) J. Biol. Chem. 279, 6235-6243). To confirm the assigned structures, LC-MS measurements with collision induced dissociation (LC-MS/MS) were performed on insulin fragments. The results of the LC-MS/MS analyses agreed excellently with the assignments. The study showed that arginine, tyrosine, and lysine residues were very reactive. However, eight theoretically reactive residues did not show detectable modifications, probably because of their low intrinsic reactivity, inaccessibility, or both. The asparagine, glutamine, and histidine residues were not converted in insulin. The N-termini of insulin were partly converted to the expected imidazolidinone adducts, indicating that the protein conformation affects the accessibility and reactivity of these residues. In conclusion, this study shows that, based on our current insights in the chemistry of the reactions between proteins and formaldehyde, we are able to elucidate the location and nature of formaldehyde induced modifications in a small protein. The approach followed in this study may be generally applicable to larger formaldehyde-treated proteins, such as toxoids used in vaccines. PMID- 16704223 TI - Receptor-mediated hepatic uptake of M6P-BSA-conjugated triplex-forming oligonucleotides in rats. AB - Excessive production of extracellular matrix, predominantly type I collagen, results in liver fibrosis. Earlier we synthesized mannose 6-phosphate-bovine serum albumin (M6P-BSA) and conjugated to the type I collagen specific triplex forming oligonucleotide (TFO) for its enhanced delivery to hepatic stellate cells (HSCs), which is the principal liver fibrogenic cell. In this report, we demonstrate a time-dependent cellular uptake of M6P-BSA-33P-TFO by HSC-T6 cells. Both cellular uptake and nuclear deposition of M6P-BSA-33P-TFO were significantly higher than those of 33P-TFO, leading to enhanced inhibition of type I collagen transcription. Following systemic administration into rats, hepatic accumulation of M6P-BSA-33P-TFO increased from 55% to 68% with the number of M6P per BSA from 14 to 27. Unlike 33P-TFO, there was no significant decrease in the hepatic uptake of (M6P)20-BSA-33P-TFO in fibrotic rats. Prior administration of excess M6P-BSA decreased the hepatic uptake of (M6P)20-BSA-33P-TFO from 66% to 40% in normal rats, and from 60% to 15% in fibrotic rats, suggesting M6P/insulin-like growth factor II (M6P/IGF II) receptor-mediated endocytosis of M6P-BSA-33P-TFO by HSCs. Almost 82% of the total liver uptake in fibrotic rats was contributed by HSCs. In conclusion, by conjugation with M6P-BSA, the TFO could be potentially used for the treatment of liver fibrosis. PMID- 16704224 TI - Development and properties of beta-glucuronide linkers for monoclonal antibody drug conjugates. AB - A beta-glucuronide-based linker for attaching cytotoxic agents to monoclonal antibodies (mAbs) was designed and evaluated. We employed the cytotoxic auristatin derivatives MMAE (1a) and MMAF (1b) and doxorubicin propyloxazoline (DPO, 2) to give the beta-glucuronide drug-linkers 9a, 9b, and 17, respectively. Cysteine-quenched derivatives of 9b and 17 were determined to be substrates for E. coli beta-glucuronidase, resulting in facile drug release. The beta glucuronide MMAF drug-linker 9b was highly stable in rat plasma with an extrapolated half-life of 81 days. Each drug-linker when conjugated to mAbs c1F6 (anti-CD70) and cAC10 (anti-CD30) gave monomeric antibody-drug conjugates (ADCs) with as many as eight drugs per mAb and had high levels of immunologically specific cytotoxic activity on cancer cell lines. cAC10-9a displayed pronounced antitumor activity in a subcutaneous Karpas 299 lymphoma tumor model. A single dose treatment led to cures in all animals at the 0.5 mg/kg dose level and above, and the conjugate was well tolerated at 100 mg/kg. In mice with subcutaneous renal cell carcinoma xenografts, the MMAF conjugate c1F6-9b was tolerated at 25 mg/kg and efficacious at 0.75 mg/kg. These results demonstrate that the beta glucuronide linker system is an effective strategy for targeting cytotoxic agents providing ADCs with high degrees of efficacy at well-tolerated doses. PMID- 16704225 TI - Synthetic modification of silica beads that allows for sequential attachment of two different oligonucleotides. AB - We developed a simple and elegant synthesis strategy that enables us to attach controlled (equimolar) amounts of two different oligonucleotides onto one silica bead. The method involves addition of orthogonally protected lysine followed by activation and derivatization of each amino group with a different moiety. This sequential oligonucleotide attachment enables the use of a combinatorial scheme to generate millions of bead types, each characterized by its two oligo tags. (In our randomly assembled arrays each bead type can then be identified by a series of hybridizations of fluorescently labeled decoder oligos to the address tags.) To demonstrate feasibility of such a scheme we created over 1000 bead types, which were characterized by their two oligo tags. The method enables genotyping or gene expression assays at multiplex levels of hundreds of thousands to millions. PMID- 16704226 TI - Targeted liposomes: convenient coupling of ligands to preformed vesicles using "click chemistry". AB - An efficient and convenient chemoselective conjugation method based on "click chemistry" was developed for coupling ligands to the surface of preformed liposomes. It can be performed under mild conditions in aqueous buffers; the use of a water soluble Cu(I) chelator, such as bathophenanthrolinedisulfonate, was essential to obtain good yields in reasonable reaction times. A model reaction was achieved in which, in a single step, an unprotected alpha-D-mannosyl derivative carrying a spacer arm functionalized with an azide group was conjugated to the surface of vesicles presenting a synthetic lipid carrying a terminal alkyne function. When liposomes composed of saturated phospholipids were used, the reaction conditions developed in the present work did not damage the membranes as measured by the absence of leakage of entrapped 5,6 carboxyfluorescein. Moreover, as assessed by agglutination experiments using concanavalin A, the mannose residues were perfectly accessible on the surface of the targeted vesicles. PMID- 16704227 TI - Synthesis of building blocks for solid-phase introduction of diethylenetriaminepentaacetic acid (DTPA) to oligonucleotides and oligopeptides. AB - Synthesis of building blocks that allow site-specific incorporation of diethylenetriaminepentaacetic acid (DTPA) to oligonucleotides and oligopeptides using phosphoramidite and Fmoc chemistries, respectively, is described. PMID- 16704228 TI - 1H NMR spectroscopy as a tool for determining the composition of poly(hydroxyethyl-L-asparagine)-coated liposomes. AB - Liposomes coated with the poly(amino acid) poly(hydroxyethyl-L-asparagine) (PHEA) show long-circulation properties comparable to the frequently used PEG-liposomes. The pharmacokinetic characteristics of long-circulating liposomes are dependent on the density of the shielding polymer on the liposome surface. Therefore, it is necessary to know the exact composition of the liposomes including the amount of coating polymer present on the liposome surface. In this study, a 1H NMR method to establish the composition of liposomes coated with PHEA was developed and validated. PMID- 16704229 TI - Planar catechin analogues with alkyl side chains: a potent antioxidant and an alpha-glucosidase inhibitor. AB - Planar catechin analogues having various alkyl side chain lengths were synthesized, and their remarkable antioxidative abilities and alpha-glucosidase inhibitory activities are shown. PMID- 16704230 TI - Releasable luciferin-transporter conjugates: tools for the real-time analysis of cellular uptake and release. AB - The design, synthesis, and evaluation of conjugates of arginine-rich transporters and luciferin are described that release luciferin only after entry into cells that are stably transfected with luciferase. Each molecule of free luciferin that is released after entry generates a photon that can be measured allowing for real time quantification of uptake and release in cells. The process provides a method to assay uptake and release of free luciferin as a function of variations in the releasable linker and in the transporter. PMID- 16704231 TI - Probing organic field effect transistors in situ during operation using SFG. AB - In this communication, we report results obtained using surface-sensitive IR+Visible Sum Frequency Generation (SFG) nonlinear optical spectroscopy on interfaces of organic field effect transistors during operation. We observe remarkable correlations between trends in the surface vibrational spectra and electrical properties of the transistor, with changes in gate voltage (VG). These results suggest that field effects on electronic conduction in thin film organic semiconductor devices are correlated to interfacial nonlinear optical characteristics and point to the possibility of using SFG spectroscopy to monitor electronic properties of OFETs. PMID- 16704232 TI - Direct synthesis of B-C-N single-walled nanotubes by bias-assisted hot filament chemical vapor deposition. AB - Direct synthesis of large-scale ternary boron carbonitride single-walled nanotubes (BCN-SWNTs) via a bias-assisted HFCVD process was presented. The BCN SWNTs were grown over the powdery Fe-Mo/MgO catalyst by using CH4, B2H6, and ethylenediamine vapor as the reactant gases. As high as 16 atom % nitrogen can be incorporated within the nanotube shells, with the boron content in the range of 2 4 atom %. The ternary covalent bonding nature of the BCN-SWNTs was well characterized, and the B, C, and N elemental maps were clearly imaged by energy filtered transmission electron microscopy. PMID- 16704233 TI - A chiral Ag-based catalyst for practical, efficient, and highly enantioselective additions of enolsilanes to alpha-ketoesters. AB - A Ag-based chiral catalyst promotes efficient and highly enantioselective aldol additions of ketone-derived enolsilanes to alpha-ketoesters in the presence of a readily available amino acid-based ligand and commercially available AgF2. alpha Ketoester substrates may bear alkyl, alkenyl, and aryl substituents; reactions proceed to >98% conversion to afford the desired tertiary alcohols in 61->98% isolated yield and 60-96% ee. In contrast to previously reported approaches, highest enantioselectivities are observed with sterically demanding substrates, and reactions can be carried out in undistilled solvent, in air with as little as 1 mol % catalyst. PMID- 16704234 TI - Contrast agents for magnetic resonance imaging synthesized with ring-opening metathesis polymerization. AB - A monomer for ring-opening metathesis polymerization (ROMP) has been developed that also functions as a portion of a GdIII chelating moiety for a magnetic resonance imaging contrast agent. An increase in per GdIII relaxivity was shown upon transition from monomer to polymer. Additionally, extremely large molecular relaxivities were achieved through incorporation of multiple GdIII ions per polymer. The nature of ROMP-derived polymers allows for functionalization of the monomer units and termini through orthogonal chemistry. This strategy is the basis for a new class of highly sensitive, targeted imaging agents. PMID- 16704235 TI - Kinetic resolution of 2-oxazolidinones via catalytic, enantioselective N acylation. AB - Kinetic resolution of racemic 2-oxazolidinones via catalytic, enantioselective N acylation has been achieved for the first time and with outstanding selectivities. PMID- 16704236 TI - Inorganic single wall nanotubes of SbPS(4-x)Se(x) (0 < or = x < or = 3) with tunable band gap. AB - A new ternary member, SbPS4, has been added to the growing inorganic nanotube family. This material naturally forms bundles of long, single wall nanotubes. PMID- 16704237 TI - Formation and characterization of polyglutamate core-shell microspheres. AB - The need for organ-targeted delivery of drugs and imaging agents creates an interest in biocompatible, biodegradable vesicles. We make protein microspheres using high-intensity ultrasound; these microspheres have a protein shell and a hydrophobic interior, making them ideal for delivering hydrophobic materials. We have previously shown that various proteins, e.g., bovine serum albumin (BSA), form a microsphere shell stabilized by interprotein cross-linking of cysteine residues. In this study, polyglutamate was used to form core-shell microspheres at slightly basic pH using sonication. These particles are smaller than our previous protein microspheres and are stable under conditions encountered in vivo. The stability of polyglutamate microspheres appears to be due to hydrogen bonding networks and not covalent cross-linking. PMID- 16704238 TI - Efficient energy transfer from peripheral chromophores to the self-assembled zinc chlorin rod antenna: a bioinspired light-harvesting system to bridge the "green gap". AB - An artificial light-harvesting rod aggregate based on zinc chlorin and covalently linked naphthalene bisimide chromophore has been realized by self-assembly. Efficient energy transfer (phiET >/= 0.99) takes place upon excitation at 620 nm from peripheral naphthalene bisimides to the zinc chlorin rod aggregate backbone. The appended naphthalene bisimide dyes improve the total LH efficiency of the rod aggregate by 26%. Thus, the present bioinspired antenna system is promising for application in nanodevices for the effective utilization of solar energy by bridging the "green gap". PMID- 16704239 TI - Quadruply hydrogen bonded cytosine modules for supramolecular applications. AB - In view of the range of properties required from supramolecular materials, there is clearly a need for new strong quadruple hydrogen bonded modules, which can be used in polymer or copolymer synthesis via the self- or hetero-association of complimentary units. A cytosine-based module has been prepared for supramolecular applications using a straightforward synthetic approach. The cytosine module was designed such that it does not undergo tautomeric changes observed with ureidopyrimidinones. The cytosine module was capable of forming quadruple hydrogen bonded assemblies both in solution and in the solid state, and the structure of the dimeric self-assembled unit was confirmed by single-crystal X ray and solution NMR techniques. The dimerization constant was estimated to be greater than 9 x 106 M-1 in deuterated benzene. The capacity of the cytosine based module to strongly hetero-associate with the ureidopyrimidinone module was demonstrated, and a supramolecular polymer of a bifunctional unit incorporating the cytosine module and PEG-based linker was described. PMID- 16704240 TI - Photocatalytic oxidation of hydrocarbons by a bis-iron(III)-mu-oxo Pacman porphyrin using O2 and visible light. AB - The C-H bonds of hydrocarbons are oxidized catalytically by the electron deficient Pacman porphyrin, (DPDF)Fe2O, using visible light and molecular oxygen as the terminal oxidant and oxygen atom source. The photocatalytic reactions proceed under mild conditions (ambient temperature and pressure) without the need for an external co-reductant. PMID- 16704241 TI - Development of a ratiometric fluorescent zinc ion probe in near-infrared region, based on tricarbocyanine chromophore. AB - Novel ratiometric fluorescent probes for Zn2+ in the near-infrared region, based on a tricarbocyanine chromophore, have been designed, synthesized, and evaluated. Upon addition of Zn2+, a 44 nm red shift of the absorption maximum was observed, which indicates that this probe could work as a ratiometric probe for Zn2+. This change is due to the difference in the electron-donating ability of the amine substituent before and after reaction with Zn2+. This fluorescence modulation of amine-substituted tricarbocyanines should be applicable to dual-wavelength measurement of various biomolecules or enzyme activities. PMID- 16704242 TI - One-step one-phase synthesis of monodisperse noble-metallic nanoparticles and their colloidal crystals. AB - A variety of metallic nanoparticles with a narrow size distribution have been synthesized in a facile one-phase method in which amine-borane complexes are applied as reducing agents. It is particularly striking that large colloidal crystals with sizes up to tens of micrometers can directly form from the reaction mixtures without any further treatment. By using the synthetic route described, large-scale syntheses of both mono- and alloyed metallic nanoparticles with a narrow size distribution can be easily achieved. PMID- 16704243 TI - Separation of semiconducting from metallic carbon nanotubes by selective functionalization with azomethine ylides. AB - A mild and efficient method for the functionalization of SWNTs by cycloaddition of azomethine ylides derived from trialkylamine-N-oxides is described. Selective reaction of semiconducting carbon nanotubes was achieved by preorganizing the starting N-oxides on the nanotube surface prior to generating the reactive ylides. Separation of met-SWNTs from functionalized sem-SWNTs was successfully accomplished by inducing solubilization of sem-SWNTs in the presence of lignoceric acid. PMID- 16704244 TI - Ferroelectric metal-organic framework with a high dielectric constant. AB - Hydrothermal reaction of (l)-N-(4'-cyanobenzy)-(S)-proline with CdCl2 as a Lewis acid catalyst and NaN3 gives colorless block compound 1, in which 1 displays a complicated 3D framework. Ferroelectric and dielectric property measurements reveal that 1 exhibits physical properties comparable to that of a typical ferroelectric compound with a dipole relaxation process and a dielectric constant of ca. 38.6 that makes it, by definition, a high dielectric material. PMID- 16704245 TI - A biomimetic "polysoap" for single-walled carbon nanotube dispersion. AB - As-synthesized single-walled carbon nanotubes (SWNTs) are bundled mixtures of different species. The current challenge in the field of carbon nanotube research lies in the processing and separation of SWNTs, which first require efficient dispersion of individual SWNTs in solvents. We report DNA-mimicking polysoap surfactants that disperse SWNTs in aqueous solutions more effectively than DNA. The polysoaps are synthesized by functionalizing the side chain of poly(styrene alt-maleic acid) with aminopyrene. The synthetic nature of the polysoap opens a new approach to further optimization of not only SWNT dispersion efficiency but also multi-functional SWNT dispersing surfactant. PMID- 16704246 TI - Direct crystallographic observation of a coordinatively unsaturated transition metal complex in situ generated within a self-assembled cage. AB - In situ crystallographic observation of an extremely labile coordinatively unsaturated transition-metal complex, Cp'Mn(CO)2 (Cp' = methylcyclopentadienyl), was achieved by generating and trapping it within the confined cavity of a self assembled cage in the crystalline state at a low temperature. The labile target Cp'Mn(CO)2 was formed by the photodissociation of the CO ligand from stable Cp'Mn(CO)3, which was accommodated within the cavity of the coordination cage. Being trapped in the cage at a low temperature, Cp'Mn(CO)2 showed sufficient stability to be analyzed by X-ray crystallography. The diffraction study revealed that the 16-electron unsaturated Mn complex adopted a pyramidal geometry rather than a planar geometry. PMID- 16704247 TI - Vertical array growth of small diameter single-walled carbon nanotubes. AB - A hot filament chemical vapor deposition method has been developed to grow vertical array single-walled carbon nanotubes (SWNTs). In this study, a hot filament (temperature greater than 2000 degrees C) was used to activate gas mixtures of hydrogen and carbon containing species at sub-atmospheric pressures. Silicon substrates decorated with islands of iron were directly inserted into a preheated furnace in which a hot filament is activating the gas. Vertical arrays of SWNTs are produced with diameters ranging from 0.78 to 1.6 nm. The samples were characterized with Raman and fluorescence spectroscopy and SEM and TEM microscopy. PMID- 16704248 TI - Polymer-coated ferromagnetic colloids from well-defined macromolecular surfactants and assembly into nanoparticle chains. AB - A novel synthetic route to polymer-coated ferromagnetic colloids of metallic cobalt has been developed. Well-defined end-functional polystyrenes were synthesized using controlled radical polymerization and used as surfactants in the thermolysis of dicobaltoctacarbonyl to afford uniform ferromagnetic nanoparticles. The presence of the polymer shell enabled prolonged colloidal stability of dispersions in a wide range of organic solvents and formed glassy encapsulating coatings around ferromagnetic cores in the solid state. These polymer-coated colloids assembled into robust, micron-sized nanoparticle chains when cast onto supporting surfaces due to dipolar associations of magnetic cores. Hierarchical assemblies were also prepared by blending polystyrene-coated cobalt colloids with larger silica beads. PMID- 16704249 TI - Synthesis of degradable model networks via ATRP and click chemistry. AB - A simple scheme involving atom transfer radical polymerization (ATRP) from a bifunctional initiator, conversion of the bromine end groups of the resulting telechelic polymer to azides, and cross-linking of this azido-telechelic macromonomer with multi-acetylene functionalized small molecules via copper catalyzed azide-alkyne cycloaddition was employed to prepare the first tert-butyl acrylate model networks. This general scheme is wide in scope, enabling synthesis of model networks possessing defined pore size from any monomer polymerizable by ATRP. Introduction of an olefin moiety into the ATRP initiator enabled degradation of the materials by ozonolysis to yield star polymer products bearing three or four arms depending on which cross-linker was employed in the parent network. Size-exclusion chromatography of the ozonolysis products confirmed the pore size of the parent network and yielded insight into the number of unreacted functionalities. Model networks derived from a trifunctional alkyne were found to be more completely cross-linked than those derived from a tetrafunctional alkyne, presumably due to less steric hindrance in the former system. PMID- 16704250 TI - Pentalenolactone biosynthesis. Molecular cloning and assignment of biochemical function to PtlH, a non-heme iron dioxygenase of Streptomyces avermitilis. AB - The hydroxylase encoded by the ptlH (SAV2991) gene from the pentalenolactone gene cluster of Streptomyces avermitilis was cloned by PCR and expressed in Escherichia coli as an N-terminal His6-tag protein. Incubation of recombinant PtlH with (+/-)-1-deoxypentalenic acid (5) in the presence of Fe(II), alpha ketoglutarate, and O2 gave (-)-11beta-hydroxy-1-deoxypentalenic acid (8), whose structure and stereochemistry were determined by a combination of 1H, 13C, COSY, HMQC, HMBC, and NOESY NMR. The steady-state kinetic parameters were kcat = 4.2 +/ 0.6 s-1 and Km (5) = 0.57 +/- 0.19 mM. 8 is a new intermediate in the conversion of the sesquiterpene pentalenene (3) to pentalenolactone (1). PMID- 16704251 TI - Porphyrin synthesis in water provides new expanded porphyrins with direct bipyrrole linkages: isolation and characterization of two heptaphyrins. AB - The use of water for the porphyrin cyclization changes the products completely. Scandium-catalyzed aqueous condensation between pentafluorobenzaldehyde and pyrrole and subsequent oxidation provides novel expanded porphyrins with direct bipyrrole linkages, of which two novel heptaphyrins have been characterized by X ray analyses. PMID- 16704252 TI - Diffuse polymer interfaces in lobed nanoemulsions preserved in aqueous media. AB - Using valence electron energy loss spectroscopy (EELS) in the cryo-scanning transmission electron microscopy (STEM), we found that the polymer-polymer interface in two-phase nanocolloids of polydimethyl siloxane (PDMS) and copolymer (methyl acrylate (MA)-methyl methacrylate (MMA)-vinyl acetate (VA)) preserved in water was diffuse despite the fact that equilibrium thermodynamics indicates it should only be on the order of a few nanometers. The diffuse interface is a result of the kinetic trapping of the copolymer within the PDMS phase, and this finding suggests new nonequilibrium pathways to control interfaces during the synthesis of multicomponent polymeric nanostructures. PMID- 16704253 TI - Facile migratory insertion of a N-heterocyclic carbene into a ruthenium-carbon double bond: a new type of reaction of a NHC ligand. AB - The reaction of [RuCp(IPri)(CH3CN)2]PF6 (IPri = 1,3-bis(2,6 diisopropylphenyl)imidazol-2-ylidene) with HCCR (R = COOMe, COOEt, COMe) yields the allyl carbene complexes [RuCp(=C(R)-eta3-CHC(R)CH-IPri)]PF6. This conversion involves selective head-to-tail coupling of two alkynes and an unusual migratory insertion of the N-heterocyclic carbene into the ruthenium-carbon double bond of a ruthenacyclopentatriene intermediate. PMID- 16704254 TI - Scanning tunneling spectroscopy in an ionic liquid. AB - Molecular redox levels can be used to modulate tunneling currents through single or small numbers of molecules and induce molecular electronic device function. While most of these devices require cryogenic conditions, room temperature operation has been demonstrated by using electrochemical gating in aqueous environments. The latter have, however, serious shortcomings with a view on their relatively high volatility and narrow stability ranges in terms of potential. Here we report the first-time use of an ionic liquid, 1-butyl-3 methylimidazoliumhexafluorophosphate (BMI), as an electrochemical gate in a Scanning Tunneling Microscope (STM) configuration. Ionic liquids are known to have a very low vapor pressure, and accessible potential ranges are in principle large, up to 6 V. In a proof-of-principle experiment, we show how a heteroleptic redox-active Os bisterpyridine complex (Ossac) can be brought to exhibit both transistor and diode function in this novel environment at room temperature. This renders ionic liquids an attractive gating medium for configurations where back gating is difficult to implement (e.g., break-junction techniques) or experimental conditions prohibit the use of aqueous or organic electrolyte media (vacuum or high temperatures). From an applied perspective, they represent a step toward solid-state molecular electronics with electrochemical gating. PMID- 16704255 TI - Nanofibril self-assembly of an arylene ethynylene macrocycle. AB - Nanofibril structures have been fabricated from an arylene ethynylene macrocycle (AEM), which consists of a square frame corner-joined by four carbazole moieties. The fabrication was performed through a gelating process by cooling a warm, homogeneous solution in cyclohexane at high temperature (e.g., 100 degrees C) to room temperature. During the gelation, the molecules become organized, with optimal pi-pi stacking in cooperation with the side-chain association. The favorable pi-pi stacking facilitates the 1D growth of molecular assembly. PMID- 16704256 TI - A tetracoordinated rhodium aminyl radical complex. AB - A [16 + 1] valence electron configured rhodium aminyl radical complex could be synthesized and characterized in detail by pulse EPR spectroscopy and DFT calculations. The unpaired electron is delocalized over the metal center and two adjacent nitrogens. H-abstraction reactions from thiols and triethylsilane show that the spin density is predominantly localized on both nitrogens. PMID- 16704257 TI - On the origin of the stereoselectivity in the alkylation of oxazolopiperidone enolates. AB - The origin of the diastereoselective alkylation of enolates of oxazolopiperidones is studied by means of theoretical calculations and experimental assays. For the unsubstituted oxazolopiperidone, the alkylation with methyl chloride is predicted to afford mainly the exo product, a finding further corroborated from the analysis of the experimental outcome obtained in the reaction of the racemic oxazolopiperidone. However, such a preference can be drastically altered by the presence of substituents attached to the fused ring. In particular, when the angular carbon adopts an R configuration in a phenylglycinol-derived oxazolopiperidone, the presence of a phenyl ring at position 3 forces the pseudo planarity of the bicyclic lactam, and the diastereoselectivity is dictated by the internal torsional strain induced in the enolate. However, when the angular carbon adopts an S configuration, the preference for the exo alkylation stems from the intermolecular steric hindrance between the enolate and the alkylating reagent. Interestingly, the intramolecular hydrogen bond formed between the phenyl ring and the carbonyl oxygen in the enolate largely reduces the difference in stability of the two TSs compared to the unsubstituted oxazolopiperidone. PMID- 16704258 TI - Ultralow-density nanostructured metal foams: combustion synthesis, morphology, and composition. AB - The synthesis of low-density, nanoporous materials has been an active area of study in chemistry and materials science dating back to the initial synthesis of aerogels. These materials, however, are most often limited to metal oxides, e.g., silica and alumina, and organic aerogels, e.g., resorcinol/formaldehyde, or carbon aerogels, produced from the pyrolysis of organic aerogels. The ability to form monolithic metallic nanocellular porous materials is difficult and sometimes elusive using conventional methodology. Here we report a relatively simple method to access unprecedented ultralow-density, nanostructured, monolithic, transition metal foams, utilizing self-propagating combustion synthesis of novel transition metal complexes containing high nitrogen energetic ligands. During the investigation of the decomposition behavior of the high-nitrogen transition metal complexes, it was discovered that nanostructured metal monolithic foams were formed in a post flame-front dynamic assembly having remarkably low densities down to 0.011 g cm(-3) and extremely high surface areas as high as 270 m(2) g( 1). We have produced monolithic nanoporous metal foams via this method of iron, cobalt, copper, and silver metals. We expect to be able to apply this to many other metals and to be able to tailor the resulting structure significantly. PMID- 16704259 TI - Regioselective bond cleavage in the dissociative electron transfer to benzyl thiocyanates: the role of radical/ion pair formation. AB - Important aspects of the electrochemical reduction of a series of substituted benzyl thiocyanates were investigated. A striking change in the reductive cleavage mechanism as a function of the substituent on the aryl ring of the benzyl thiocyanate was observed, and more importantly, a regioselective bond cleavage was encountered. A reductive alpha-cleavage (CH(2)-S bond) was seen for cyano and nitro-substituted benzyl thiocyanates leading to the formation of the corresponding nitro-substituted dibenzyls. With other substituents (CH(3)O, CH(3), H, Cl, and F), both the alpha (CH(2)-S) and the beta (S-CN) bonds could be cleaved as a result of an electrochemical reduction leading to the formation of the corresponding substituted monosulfides, disulfides, and toluenes. These final products are generated through either a protonation or a nucleophilic reaction of the two-electron reduction-produced anion on the parent molecule. The dissociative electron transfer theory and its extension to the formation/dissociation of radical anions, as well as its extension to the case of strong in-cage interactions between the produced fragments ("sticky" dissociative electron transfer (ET)), along with the theoretical calculation results helped rationalize (i) the observed change in the ET mechanism, (ii) the dissociation of the radical anion intermediates formed during the electrochemical reduction of the nitro-substituted benzyl thiocyanates, and more importantly (iii) the regioselective reductive bond cleavage. PMID- 16704260 TI - C64H4: production, isolation, and structural characterizations of a stable unconventional fulleride. AB - Unconventional fullerenes are those smaller than C(60) or those intermediate between C(60) and C(70), which are not stable in structure as none of the unconventional fullerene isomers satisfying the "isolated-pentagon-rule" (IPR). Below we report the synthesis of a stable unconventional fullerene derivative C(64)H(4) by introducing methane in the fullerene productions with the normal Kratschmer-Huffman method. We also applied various spectroscopic measurements such as mass spectrometry, (13)C NMR, IR, UV-vis absorption spectrometry, etc. to characterize the structural and electronic properties of this molecule, revealing an unprecedented fullerene cage with a triplet of directly fused pentagons in the framework of C(64)H(4). Four hydrogen atoms are added to the carbons at vertexes of fused pentagons to allow the bond angles at these sites close to the sp(3) tetrahedral angle, which essentially release the sp(2) bond strains on the abutting-pentagon sites of C(64). Ab initio calculations were performed to explore the electronic property and simulate the (13)C NMR and IR spectra of this fulleride, which reproduced well the experimental results and confirmed the structural assignment of the C(64)H(4). PMID- 16704261 TI - Design of functional ferritin-like proteins with hydrophobic cavities. AB - Ferritin four-helix bundle subunits self-assemble to create a stable multimer with a large central hydrophilic cavity where metal ions bind. To explore the versatility of this reaction vessel, computational design was used to generate cavities with increasingly apolar surface areas inside a dodecameric ferritin like protein, Dps. Cavity mutants, in which as many as 120 surface accessible hydrophilic residues were replaced with hydrophobic amino acids, were shown to still assemble properly using size-exclusion chromatography and dynamic light scattering measurements. Wild-type Dps exhibited highly cooperative subunit folding and assembly, which was monitored by changes in Trp fluorescence and UV circular dichroism. The hydrophobic cavity mutants showed distinctly less cooperative unfolding behavior, with one mutant forming a partially assembled intermediate upon guanidine denaturation. Although the stability of Dps to such denaturation decreased with increasing apolar surface area, all proteins exhibited high melting temperatures, T(m) = 74-90 degrees C. Despite the large number of mutations, near-native ability to mineralize iron was maintained. This work illustrates the versatility of the ferritin scaffold for engineering large protein cavities with novel properties. PMID- 16704262 TI - Dendritic arrays of [Re6(mu3-Se)8]2+ core-containing clusters: exploratory synthesis and electrochemical studies. AB - The reaction between the previously reported site-differentiated cluster solvate [Re(6)(mu(3)-Se)(8)(PEt(3))(5)(MeCN)](SbF(6))(2) (1) with pyridyl-based ditopic ligands 4,4'-trimethylenedipyridine (2), 1,2-bis(4-pyridyl)ethane (3), and (E) 1,2-bis(4-pyridyl)ethene (4) afforded cluster complexes of the general formula [Re(6)(mu(3)-Se)(8)(PEt(3))(5)(L)](SbF(6))(2) (5-7), where L represents one of the pyridyl-based ligands. Reacting these cluster complex-based ligands with the fully solvated cluster complex [Re(6)(mu(3)-Se)(8)(MeCN)(6)](SbF(6))(2) (8) produced dendritic arrays of the general formula {Re(6)(mu(3)-Se)(8)[Re(6)(mu(3) Se)(8)(PEt(3))(5)(L)](6)}(SbF(6))(14) (9-11), each featuring six circumjacent [Re(6)(mu(3)-Se)(8)(PEt(3))(5)](2+) units bridged to a [Re(6)(mu(3)-Se)(8)](2+) core cluster by the pyridyl-based ligands. Electrochemical studies using a thin layer electrochemical cell revealed cluster-based redox events in these cluster arrays. For 9 (L = 2), one reversible oxidation event corresponding to the removal of 7 electrons was observed, indicating noninteraction or extremely weak interactions between the clusters. For 10 (L = 3), two poorly resolved oxidation waves were found. For 11 (L = 4), two reversible oxidation events, corresponding respectively to the removal of 1 and 6 electrons, were observed with the 1 electron oxidation event occurring at a potential 150 mV more positive than the 6 electron oxidation. These electrochemical studies suggest intercluster coupling in 11 via through-bond electronic delocalization, which is consistent with electronic spectroscopic studies of this same molecule. PMID- 16704263 TI - Synthesis, photochemistry, and electrochemistry of single-wall carbon nanotubes with pendent pyridyl groups and of their metal complexes with zinc porphyrin. Comparison with pyridyl-bearing fullerenes. AB - A soluble, functionalized Py-SWNT has been synthesized and characterized by solution (1)H and (13)C NMR, FT-Raman, and electron microscopy. Experimental data indicate that Py-SWNT has short tubes with pentyl esters at the tips and pyridyl isoxazolino units along the walls. The synthesis of Py-SWNT is based on a 1,3 dipolar cycloaddition of a nitrile oxide on the SWNT walls, similar to 1,3 dipolar cycloadditions that are common for fullerene functionalization. The resulting Py-SWNT forms a complex with a zinc porphyrin (ZnPor) in a way similar to that reported for pyridyl-functionalized [60]-fullerenes. Formation of this metal-ligand complex was firmly established by a detailed electrochemical study. However, in contrast to the behavior observed for the ZnPor/Py-C(60) complex, photochemical excitation of the complex between ZnPor/Py-SWNT does not lead to electron transfer with the generation of charge-separated states. Fluorescence and laser flash studies indicate that the main process is energy transfer from the singlet ZnPor excited state to the Py-SWNT with observation of emission from Py-SWNT. Triplet ZnPor excited-state quenching by Py-SWNT is only observed in polar solvents such as DMF, but not in benzonitrile. PMID- 16704264 TI - Weakly bound water molecules shorten single-stranded DNA. AB - In this paper, we measure the single chain elasticity of an oligomer single stranded DNA (ssDNA) in both aqueous and nonaqueous, apolar liquid environments by AFM-based single molecule force spectroscopy. We find a marked deviation between the force-extension relations recorded for the two conditions. This difference is attributed to the additional energy required to break the H-bond directed water bridges around the ssDNA chain in aqueous solutions, which are nonexistent in organic solvents. The results obtained in 8 M guanidine-HCl solution provide more evidence that water bridges around ssDNA originate the observed deviation. On the basis of the results obtained by an ab initio quantum mechanics calculation, a parameter-free freely rotating chain model is proposed. We find that this model is in perfect agreement with the experimental force extension curve obtained in organic solvents, which further corroborates our assumption. On the basis of the experimental results, it is suggested that the weak H-bonding between ssDNA and water molecules may be a precondition for stable double-stranded DNA to exist in water. PMID- 16704265 TI - Insights into the mechanism and catalysis of the native chemical ligation reaction. AB - Native chemical ligation of unprotected peptide segments involves reaction between a peptide-alpha-thioester and a cysteine-peptide, to yield a product with a native amide bond at the ligation site. Peptide-alpha-thioalkyl esters are commonly used because of their ease of preparation. These thioalkyl esters are rather unreactive so the ligation reaction is catalyzed by in situ transthioesterification with thiol additives. The most common thiol catalysts used to date have been either a mixture of thiophenol/benzyl mercaptan, or the alkanethiol MESNA. Despite the use of these thiol catalysts, ligation reactions typically take 24-48 h. To gain insight into the mechanism of native chemical ligaton and in order to find a better catalyst, we investigated the use of a number of thiol compounds. Substituted thiophenols with pK(a) > 6 were found to best combine the ability to exchange rapidly and completely with thioalkyl esters, and to then act as effective leaving groups in reaction of the peptide thioester with the thiol side chain of a cysteine-peptide. A highly effective and practical catalyst was (4-carboxylmethyl)thiophenol ('MPAA'), a nonmalodorous, water-soluble thiol. Use of MPAA gave an order of magnitude faster reaction in model studies of native chemical ligation and in the synthesis of a small protein, turkey ovomucoid third domain (OMTKY3). MPAA should find broad use in native chemical ligation and in the total synthesis of proteins. PMID- 16704266 TI - Triplet energy back transfer in conjugated polymers with pendant phosphorescent iridium complexes. AB - The nature of Dexter triplet energy transfer between bonded systems of a red phosphorescent iridium complex 13 and a conjugated polymer, polyfluorene, has been investigated in electrophosphorescent organic light-emitting diodes. Red emitting phosphorescent iridium complexes based on the [Ir(btp)(2)(acac)] fragment (where btp is 2-(2'-benzo[b]thienyl)pyridinato and acac is acetylacetonate) have been attached either directly (spacerless) or through a (CH(2))(8)- chain (octamethylene-tethered) at the 9-position of a 9-octylfluorene host. The resulting dibromo-functionalized spacerless (8) or octamethylene tethered (12) fluorene monomers were chain extended by Suzuki polycondensations using the bis(boronate)-terminated fluorene macromonomers 16 in the presence of end-capping chlorobenzene solvent to produce the statistical spacerless (17) and octamethylene-tethered (18) copolymers containing an even dispersion of the pendant phosphorescent fragments. The spacerless monomer 12 adopts a face-to-face conformation with a separation of only 3.6 A between the iridium complex and fluorenyl group, as shown by X-ray analysis of a single crystal, and this facilitates intramolecular triplet energy transfer in the spacerless copolymers 17. The photo- and electroluminescence efficiencies of the octamethylene-tethered copolymers 18 are double those of the spacerless copolymers 17, and this is consistent with suppression of the back transfer of triplets from the red phosphorescent iridium complex to the polyfluorene backbone in 18. The incorporation of a -(CH(2))(8)- chain between the polymer host and phosphorescent guest is thus an important design principle for achieving higher efficiencies in those electrophosphorescent organic light-emitting diodes for which the triplet energy levels of the host and guest are similar. PMID- 16704267 TI - 1H NMR study of the magnetic properties and electronic structure of the hydroxide complex of substrate-bound heme oxygenase from Neisseria meningitidis: influence of the axial water deprotonation on the distal H-bond network. AB - The substrate and active site residues of the low-spin hydroxide complex of the protohemin complex of Neisseria meningitidis heme oxygenase (NmHO) have been assigned by saturation transfer between the hydroxide and previously characterized aquo complex. The available dipolar shifts allowed the quantitation of both the orientation and anisotropy of the paramagnetic susceptibility tensor. The resulting positive sign, and reduced magnitude of the axial anisotropy relative to the cyanide complex, dictate that the orbital ground state is the conventional "d(pi)" (d(2)(xy)(d(xz), d(yz))(3)); and not the unusual "d(xy)" (d(2)(xz)d(2)(yz)d(xy)) orbital ground state reported for the hydroxide complex of the homologous heme oxygenase (HO) from Pseudomonas aeruginosa (Caignan, G.; Deshmukh, R.; Zeng, Y.; Wilks, A.; Bunce, R. A.; Rivera, M. J. Am. Chem. Soc. 2003, 125, 11842-11852) and proposed as a signature of the HO distal cavity. The conservation of slow labile proton exchange with solvent from pH 7.0 to 10.8 confirms the extraordinary dynamic stability of NmHO complexes. Comparison of the diamagnetic contribution to the labile proton chemical shifts in the aquo and hydroxide complexes reveals strongly conserved bond strengths in the distal H bond network, with the exception of the distal His53 N(epsilon)(1)H. The iron ligated water is linked to His53 primarily by a pair of nonligated, ordered water molecules that transmit the conversion of the ligated H-bond donor (H(2)O) to a H bond acceptor (OH(-)), thereby increasing the H-bond donor strength of the His53 side chain. PMID- 16704269 TI - Direct determination of low-dimensional structures: synchrotron X-ray scattering on one-dimensional charge-ordered MMX-chain complexes. AB - A powerful method to determine the hidden structural parameters in functional molecules has been developed. Local valence arrangements that dominate the material properties are sometimes not three-dimensionally ordered. This method that comprises diffuse X-ray scattering and resonant X-ray scattering is suitable in such cases. Using this method, we present clear evidence of the low dimensional valence arrangement in two halogen-bridged one-dimensional metal complexes, so-called MMX chains. This family allows us to control many physical and structural parameters by chemical substitution of bridging halogen, counterions, or metal ions, and one of our samples carries an unusual metallic phase. It is demonstrated with this complex that the present method makes it possible to have microscopic insight to low-dimensionally ordered systems. PMID- 16704268 TI - Self-assembly of surfactant vesicles that transform into viscoelastic wormlike micelles upon heating. AB - Unilamellar vesicles are observed to form in aqueous solutions of the cationic surfactant, cetyl trimethylammonium bromide (CTAB), when 5-methyl salicylic acid (5mS) is added at slightly larger than equimolar concentrations. When these vesicles are heated above a critical temperature, they transform into long, flexible wormlike micelles. In this process, the solutions switch from low viscosity, Newtonian fluids to viscoelastic, shear-thinning fluids having much larger zero-shear viscosities (e.g., 1000-fold higher). The onset temperature for this transition increases with the concentration of 5mS at a fixed CTAB content. Small-angle neutron scattering (SANS) measurements show that the phase transition from vesicles to micelles is a continuous one, with the vesicles and micelles coexisting over a narrow range of temperatures. The tunable vesicle-to-micelle transition and the concomitant viscosity increase upon heating may have utility in a range of areas, including microfluidics, controlled release, and tertiary oil recovery. PMID- 16704270 TI - Nucleophilic-alkylation-reoxidation: a functionalization sequence for single-wall carbon nanotubes. AB - A new reaction sequence for the chemical functionalization of single-wall carbon nanotubes (SWNTs) consisting of the nucleophilic addition of t-BuLi to the sidewalls of the tubes and the subsequent reoxidation of the intermediates t Bu(n)SWNT(n-) leading to t-Bu(n)SWNT was developed. During the formation of the t Bu(n)SWNT(n-), a homogeneous dispersion in benzene was formed due to the electrostatic repulsion of the negatively charged intermediates causing debundling. The entire reaction sequence can be repeated, and the degree of functionalization of the products (t-Bu(n))(m)SWNT (m = 1-3) increases with increasing m. Degrees of functionalization expressed as the carbon-to-addend ratio of up to 31 were reached. The reaction was studied in detail by photoelectron spectroscopy, Raman spectroscopy, and scanning tunneling microscopy (STM). The C 1s core level spectra reveal that the nucleophilic attack of the t BuLi leads to negatively charged SWNTs. Upon oxidation, this negative charge is removed. The valence band spectra of the functionalized samples exhibit a significant reduction in the pi-derived density of states. In STM, the covalently bonded t-butyl groups attached to the sidewalls have been visualized. Raman spectroscopy reveals that addition of the nucleophile to metallic tubes is preferred over the addition to semiconducting tubes. PMID- 16704271 TI - Heterodimers based on CoPt3-Au nanocrystals with tunable domain size. AB - We describe an approach to synthesize colloidal nanocrystal heterodimers composed of CoPt(3) and Au. The growth is based on the nucleation of gold domains on preformed CoPt(3) nanocrystals. It is a highly versatile methodology which allows us to tune independently the size of the two domains in each dimer by varying several reaction parameters. The statistical analysis of the distribution of the domain sizes in the dimers and the compositional mapping achieved by dark field imaging and energy dispersive spectroscopy confirm that the two domains in each dimer are indeed made of CoPt(3) and Au, respectively. Structural characterization by high-resolution transmission electron microscopy shows that the two domains, both having cubic fcc Bravais lattice, can share a common {111}, {100}, or {110} facet, depending on the size of the initial CoPt(3) seeds. The magnetization measurements evidence a ferromagnetic CoPt(3) phase with a relatively low anisotropy as a consequence of their disordered crystalline structure, regardless of the presence of a Au tip. We believe that this prototype of nanocrystal dimer, which can be manipulated under air, can find several applications in nanoscience, as the Au section can be exploited as the preferential anchor point for various molecules, while the CoPt(3) domain can be used for magnetic detection. PMID- 16704272 TI - Host-guest complexation of endohedral metallofullerene with azacrown ether and its application. AB - Complexation of endohedral metallofullerene La@C(82)-A (1) with macrocyclic compounds, such as 1,4,7,10,13,16-hexaazacyclooctadecane (2), 1,4,7,10,13,16 hexamethyl-1,4,7,10,13,16-hexaazacyclooctadecane (3), mono-aza-18-crown-6 ether (4), 18-crown-6 ether (5), and p-tert-butylcalix[n]arenes (n = 4-8, 6-10), for the first time is examined. Among them, 1 forms a complex with azacrown ethers 2 4 while accompanying the electron transfer between them. This is characteristic of endohedral metallofullerene and caused by its low reduction potential. Activation energies, DeltaG(et), for the electron transfer from 2-4 to 1 are 4.6, 2.8, and 11 kcal/mol, respectively. These small DeltaG(et) values indicate that the electron transfer from the azacrown ethers to 1 is facile in the ground state. Furthermore, the selective isolation of lanthanum endohedral metallofullerenes from the extracts of soot is accomplished by utilizing the complexation of 1 with 2. PMID- 16704273 TI - Mixed micelles of fluorinated and hydrogenated surfactants. AB - The model mixed surfactant system of sodium perfluorooctanoate and sodium decyl sulfate was carefully reexamined by a combination of nuclear magnetic resonance methods. Over a wide range of sample compositions, detailed (19)F and (1)H chemical shift data in combination with self-diffusion coefficients for the perfluorooctanoate and decyl sulfate ions are collected. All data are analyzed together in a framework that uses a minimal number of initial assumptions to extract the monomer concentrations of both surfactants and the micellar chemical shifts of (19)F and (1)H as a function of relative concentration. The main conclusion drawn from this analysis is that there exists neither complete demixing nor complete mixing on molecular or micellar levels. Instead, the experimental data favor a single type of micelles within which fluorinated surfactants are preferentially coordinated by fluorinated ones and hydrogenated surfactants by hydrogenated ones. The data are quantitatively interpreted in the framework of the first approximation of the regular solution theory (also called the quasi-chemical treatment) leading to an energy of mixing of omega = W/kT = 0.98 between the constituting surfactant types. These findings may help to resolve a long controversy about micellar mixing-demixing in this particular mixture and in its relatives. PMID- 16704274 TI - Self-assembly, structural, and retrostructural analysis of dendritic dipeptide pores undergoing reversible circular to elliptical shape change. AB - The synthesis of dendritic dipeptides (4-3,4-3,5-4)12G2-CH(2)-Boc-L-Tyr-L-Ala-OMe and (4-3, 4-3,5-4)12G2-CH(2)-Boc-D-Tyr-D-Ala-OMe is described. These dendritic dipeptides self-assemble into porous elliptical and circular columns that in turn self-organize into centered rectangular columnar and hexagonal columnar periodic arrays. The transition from porous elliptical to porous circular columns is mediated in a reversible or irreversible way by the thermal history of the sample. A method to determine the dimensions of hollow elliptical and circular columns by the reconstruction of the small-angle powder X-ray diffractograms of the centered rectangular or hexagonal columnar lattices was elaborated. This technique together with wide-angle X-ray experiments performed on aligned fibers provided access to the structural and retrostructural analysis of elliptical supramolecular pores. This procedure is general and can be adapted for the determination of the dimensions of pores of any columnar shape. PMID- 16704275 TI - Expression, purification, and characterization of a galactofuranosyltransferase involved in Mycobacterium tuberculosis arabinogalactan biosynthesis. AB - The major structural component of the cell wall of Mycobacterium tuberculosis is a lipidated polysaccharide, the mycoyl-arabinogalactan-peptidoglycan (mAGP) complex. This glycoconjugate plays a key role in the survival of the organism, and thus, enzymes involved in its biosynthesis have attracted attention as sites for drug action. At the core of the mAGP is a galactan composed of D galactofuranose residues attached via alternating beta-(1-->5) and beta-(1-->6) linkages. A single enzyme, glfT, has been shown to synthesize both glycosidic linkages. We report here the first high-level expression and purification of glfT by expression of the Rv3808c gene in Escherichia coli C41(DE3). Following a three step purification procedure, 3-7 mg of protein of >95% purity was isolated from each liter of culture. We subsequently probed the substrate specificity of glfT by evaluating a panel of potential mono- and oligosaccharide substrates and demonstrated, for the first time, that trisaccharides are better substrates than disaccharides and that one disaccharide, in which the terminal D-galactofuranose residue is replaced with an L-arabinofuranose moiety, is a weak substrate. Kinetic characterization of the enzyme using four of the oligosaccharide acceptors gave K(m) values ranging from 204 microM to 1.7 mM. Through the use of NMR spectroscopy and mass spectrometry, we demonstrated that this recombinant enzyme, like the wild-type protein, is bifunctional and can synthesize both beta (1-->6) and beta-(1-->5)-linkages in an alternating fashion. Access to purified glfT is expected to facilitate the development of high-throughput assays for the identification of inhibitors of the enzyme, which are potential antituberculosis agents. PMID- 16704276 TI - Spontaneous transformation of CdTe nanoparticles into angled Te nanocrystals: from particles and rods to checkmarks, X-marks, and other unusual shapes. AB - CdTe nanoparticles spontaneously transform into the branched Te nanocrystals with the unique, highly anisotropic shape of checkmarks after partial removal of the stabilizers of L-cysteine. The Te checkmarks are made in a relatively high yield and uniformity; the length of the arms is ca. 150 nm, whereas the angle between the arms is 74 degrees . Subsequent growth of the particle yields mothlike nanocrystals retaining geometrical anisotropy. Unlike the previous synthesis methods of branched nanocrystals, they are formed via a merger of individual rod shaped crystallites. High-energy crystal faces on their apexes act as the sticky points causing the particles to join in the ends. This is the first demonstration of spontaneous transformation of binary semiconductor particles into highly anisotropic nanocolloids in an angled conformation. The end reactivity of starting Te rods can be used both for bottom-up fabrication of nanoscale electronics and relatively safe and nontoxic method of synthesis of Te-based optical and other materials. PMID- 16704277 TI - Micropores in crystalline dipeptides as seen from the crystal structure, He pycnometry, and 129Xe NMR spectroscopy. AB - Eight crystalline dipeptides were studied: AV (Ala-Val), VA (Val-Ala), AI (Ala Ile), VV (Val-Val), IA (Ile-Ala), IV (Ile-Val), VI (Val-Ile), and LS (Leu-Ser) (all LL isomers). The first seven form an isostructural series (space group P6(1)), whereas LS has a different structure (P6(5)). All structures display H bonded tubular assemblies of the dipeptide molecules resulting in open ultramicropores in the form of isolated one-dimensional (1D) channels. The total porosity of the materials ranges from 4 to 12% (micropore volume from 0.04 to 0.12 cm(3)/g). Calculations based on the crystal structures, He pycnometry, and solid-state (129)Xe NMR methods were used to obtain a comprehensive description of the geometry and properties of the micropores. The following order was established for the channel diameter: AV > VA > AI > VV > IA > IV > VI, with >5 A for AV and <4 A for VI; LS is close to AI. The observed sorption behavior cannot be described adequately based on the crystal structure and can only be understood if one takes into account the dynamics of the host matrix. The pores are chiral, with the center of the channel describing a right-handed helix (left-handed for LS). The following order was established for the channel helicity: VA > IA > IV > AV approximately AI approximately VV > VI > LS, with a helix diameter of approximately 2 A for VA, IA, and IV and approximately 1 A or less for the remaining dipeptides. A comparison of the dipeptides studied with other supramolecular materials is given and the potential for applications is discussed. PMID- 16704279 TI - Conformational stability of the propylene oxide-water adduct: direct spectroscopic detection of O-H...O hydrogen bonded conformers. AB - The 1:1 molecular adduct of propylene oxide and water (PO-H(2)O) was studied using Fourier transform microwave spectroscopy and high level ab initio methods. Two distinct structural conformers with the water molecule acting as a proton donor were detected experimentally: one with the water on the same side as the methyl group with respect to the ether ring, i.e., syn-PO-H(2)O, the other with the water molecule binding to the O-atom from the opposite side of the methyl group, i.e., anti-PO-H(2)O. The nonbonded hydrogen is entgegen to the ether ring in both conformers. Rotational spectra of four isotopic species, namely PO-H(2)O, PO-DOH, PO-HOD, and PO-D(2)O, were recorded for the two conformers. The hydrogen bond parameters: r(O(epoxy)...H), angle(ring-O(epoxy)...H), and angle(O(epoxy)...H-O) are 1.908 A, 112 degrees, and 177 degrees for syn-PO-H(2)O, and 1.885 A, 104.3 degrees, and 161.7 degrees for anti-PO-H(2)O, respectively. The experimental results suggest that the hydrogen bond in syn-PO-H(2)O is stronger and the monomer subunits are more rigidly locked in their positions than in the ethylene oxide-water adduct. The stabilizing effect of the methyl group to the intermolecular hydrogen bond is discussed in terms of the experimentally estimated binding energies, the structural parameters, and the ab initio calculations. PMID- 16704278 TI - Sequential Ru-Pd catalysis: a two-catalyst one-pot protocol for the synthesis of N- and O-heterocycles. AB - An atom economic, selective, and highly practical two-metal one-pot synthesis of heterocycles has been developed that efficiently affords enantio- and diastereopure N- and O-heterocyclic products. Furthermore, use of a chiral catalyst in the two-metal procedure allows formation of all possible diastereomers, even those that are traditionally difficult to access via cyclization routes due to thermodynamics. Interestingly, the nature of the enantiodiscriminating event differs between the use of amine versus alcohol nucleophiles. The method also affords heterocyclic products that are synthetically useful intermediates. Through the Z-vinylsilane a variety of stereodefined trisubstituted olefin products can be accessed including several all-carbon motifs. Finally, the utility of these heterocyclic products in total synthesis is demonstrated through concise syntheses of a kainoid intermediate, a constituent of oil of rose, and the ring B portion of bryostatin, a potent chemotherapeutic. PMID- 16704280 TI - Participate in school nursing research: return those questionnaires. PMID- 16704281 TI - Helping teen mothers succeed. AB - Because the success of teen mothers is enhanced by completing high school, school districts should give high priority to supporting teen mothers to remain in school and to graduate. This article reviews the literature on the educational attainment of these students, their school aspirations, and the policies affecting their education. Although teens often begin mothering with a range of educational and social disadvantages, many teen mothers recommit to school to enhance their future opportunities. Unfortunately, rising school aspirations among teen mothers often are undermined by competing demands and the lack of consistent family and school support. School nurses can support teen mothers' aspirations and contribute to their long-term success by linking them to resources and advocating for policies and practices that promote high school graduation. PMID- 16704282 TI - Encouraging sun safety for children and adolescents. AB - The rise in the number of cases of skin cancers, both melanomas and nonmelanomas, has prompted increased awareness and educational efforts to limit sun exposure. Because 80% of lifetime sun exposure occurs before the age of 18, educating parents and adolescents to incorporate sun-protective behaviors into daily routines is particularly important. Education of parents is essential to establishing healthy behavior patterns in children. Educational interventions are recommended that encourage the following: using sunscreen, wearing hats with wide brims and clothing that blocks or absorbs ultraviolet rays, using sunglasses that block ultraviolet radiation, and seeking shade. Continued efforts are important through adolescence to maintain the established behaviors. School nurses are in a position to affect the health education curriculum, as well as school policies that promote sun safety behaviors. PMID- 16704283 TI - School health nursing services role in education: the No Child Left Behind Act of 2001. AB - Public Law 107-110, the No Child Left Behind Act of 2001, represents the most sweeping reauthorization of the Elementary and Secondary Education Act since it was enacted in 1965. The goal of the No Child Left Behind Act is to improve the academic achievement of all American students so that they all meet their own state's standards for competence by 2014. The law focuses on closing the achievement gap that exists among the economically disadvantaged, those with limited English proficiency, racial/ethnic minorities, and students receiving special education services. The No Child Left Behind Act has created both support and controversy in education circles. Although the No Child Left Behind Act does not legislate the practice of school nursing, as professional staff working in schools, school nurses are affected indirectly by the heightened emphasis on accountability for achievement in public education. School nurses actively promote and support achievement for all students in specific ways and help to provide a safe school environment. PMID- 16704284 TI - Medication administration practices in Pennsylvania schools. AB - As a result of various health concerns, children are receiving an increased number of medications while at school. In Pennsylvania, the School Code mandates a ratio of 1 certified school nurse to 1,500 students, which may mean that 1 school nurse is covering 3-5 buildings. This implies that unlicensed personnel are administering medications, a violation of licensing laws in Pennsylvania. The purpose of this study was to determine the policies and practices that Pennsylvania public schools have in place regarding medication administration. The sample consisted of 314 state-certified school nurses who returned a 71 question survey. Findings indicated that school nurses are very concerned about issues related to medication administration. Additional findings identified the lack of standing orders to administer over-the-counter medications, the increase in medication errors when the school nurse had responsibility for multiple buildings, and the lack of understanding of the ramifications of the Nurse Practice Act on school nursing practice in regard to delegation. Recommendations for practice include development of detailed policies and procedures and collaboration among all stakeholders in the development of policies that address legal issues. PMID- 16704285 TI - Lack of follow-up exams after failed school vision screenings: an investigation of contributing factors. AB - Programs to facilitate professional eye exams after failed school vision screenings often are based on the assumption that funding and access to services are major obstacles to care. Despite such programs, many children do not receive professional exams. The purpose of this study was to identify additional barriers to follow-up eye care. School nurses in an urban, midwestern public school district identified elementary school students who had not received follow-up eye exams after failed school vision screenings. Parents of these students were interviewed during the summer to determine financial, logistical, social/family, and perceptual barriers to care. Family issues, parental perceptions of vision problems, and difficulty planning ahead were found to be significant factors. Strategies to increase follow-up compliance and recommendations for overcoming barriers to care were also identified. PMID- 16704286 TI - Young caregivers: effect of family health situations on school performance. AB - Recognition, assessment, education, support, and research of young caregivers is yet to be well established throughout the United States. Studies show that as a result of family caregiving, employees miss work, arrive late, have telephone interruptions, and pass up promotions. Education is essentially the work-life of a student. Research among 12,681 public school students in Palm Beach County, Florida, shows that more than 1 in 2 middle and high school youth (6,210) have the dual role of young caregiver and student; one third of all students (67.1% of young caregivers) miss school/after school activities, do not complete homework, and/or are interrupted in their studying-their work-life. As demonstrated in the United Kingdom, assessment, education, and support of young caregivers in school and in the community can improve education outcomes. Proactive school nurses have the opportunity to identify, assess, and promote the well-being and academic success of U.S. student-caregivers. PMID- 16704287 TI - Asthma education for rural school nurses: resources, barriers, and outcomes. AB - This paper describes a school-based asthma education program for rural elementary school nurses. The program was designed to teach school nurses in 7 rural counties in Maryland how to implement and to reinforce asthma management behaviors in children with asthma and their caregivers. Rural nurses who participated in this program increased their mean asthma knowledge scores more than nurses who did not take the program. The program also increased self efficacy among intervention school nurses, but the difference in self-efficacy between intervention and control nurses was not statistically significant at follow-up. No effects on documentation or communication behaviors were noted. Only 25% of the nurses reported an interest in implementing future asthma educational programs for children with asthma. This study indicates the importance of understanding the unique characteristics of rural school nurses, the resources they need, and the barriers and challenges they face in their practice. PMID- 16704288 TI - Comparison of health-related quality of life among 10- to 12-year-old children with chronic illnesses and healthy children: the parents' perspective. AB - The purpose of this study was to evaluate mothers' and fathers' perception of their child's health-related quality of life (HRQOL) among 10- to 12-year-old Icelandic children with or without chronic health condition or illness. A total of 912 Icelandic parents (510 mothers and 402 fathers) and 480 children (209 boys and 271 girls) participated in the study. The Icelandic fathers of children who visited the school nurse over a period of 1 week perceived HRQOL of their children to be significantly lower than the mothers. A gender difference was found between the parents; mothers perceived their children to have significantly higher school functioning than fathers. Both mothers and fathers of children with chronic health condition/illness perceived their children to have significantly lower HRQOL than did the parents of children without a chronic health condition. The findings underline the importance for school nurses to develop and test interventions for school-age children with chronic health conditions or illnesses. PMID- 16704289 TI - Animal experimentation: should the three Rs be abandoned? PMID- 16704290 TI - The FRAME alternatives laboratory database. 1. In vitro basal cytotoxicity determined by the Kenacid blue total protein assay. AB - A database of over 280 chemicals has been compiled by using a mouse 3T3-L1 fibroblast-like cell line in exponential growth, exposed to chemicals for 72 hours in a 96-well tissue culture plate format, and determining cell number via the Kenacid blue (KB) assay for total protein. Ranking the chemicals according to their basal cytotoxicity, expressed as the concentration (mM) that inhibits increase in total cellular protein over 72 hours by 50% (the ID50 value) shows a wide range of ID50 values, from 0.00003 mM to 10,096 mM. This information includes the results for MEIC chemicals 1-50, and we have now added basal cytotoxicity data for 23 of the next 25 MEIC chemicals. When the neutral red uptake (NRU) assay was performed with the same cell cultures, before the KB assay, very similar indications of basal cytotoxicity were obtained. Comparisons between the results with 3T3-L1 cells and with a human fibroblast-like cell line, BCL-D1 showed a significant difference in order of magnitude of the ID50 value for only 5 of 52 chemicals. However, there was a difference in ID50 value of more than one order of magnitude for 8 of 24 chemicals tested with an undifferentiated teratocarcinoma cell line, F9. PMID- 16704291 TI - Designing validation studies more efficiently according to the modular approach: retrospective analysis of the EPISKIN test for skin corrosion. AB - It is claimed that the modular approach to validation, which involves seven independent modules, will make the assessment of test validity more flexible and more efficient. In particular, the aspects of between-laboratory variability and predictive capacity are formally separated. Here, the main advantage of the approach is to offer the opportunity for reduced labour, and thus to allow study designs to be more time efficient and cost effective. The impact of this separation was analysed by taking the ECVAM validation study on in vitro methods for skin corrosivity as an example of a successful validation study - two of its methods triggered new OECD test guidelines. Lean study designs, which reduced the number of tests required by up to 60%, were simulated with the original validation data for the EPISKIN model. By using resampling techniques, we were able to demonstrate the effects of the lean designs on three between-laboratory variability measures and on the predictive capacity in terms of sensitivity and specificity, in comparison with the original study. Overall, the study results, especially the levels of confidence, were only slightly affected by the lean designs that were modelled. It is concluded that the separation of the two modules is a promising way to speed-up prospective validation studies and to substantially reduce costs, without compromising study quality. PMID- 16704292 TI - Three Rs Approaches in Marine Biotoxin Testing. The Report and Recommendations of a joint ECVAM/DG SANCO Workshop (ECVAM Workshop 54). PMID- 16704293 TI - TGN1412: time to change the paradigm for the testing of new pharmaceuticals. AB - Clinical studies in human volunteers are an essential part of drug development. These studies are designed to account for possible differences between the effects of pharmaceutical products in preclinical studies and in humans. However, the tragic outcome of the recent Phase 1 clinical trial on TGN1412 casts considerable doubt over the relevance of this traditional drug development paradigm to the testing of therapeutic agents for human use. The role of alternatives to animal testing is considered, and a series of recommendations are made, which could ensure that clinical trials are well informed and based on the most relevant scientific information. PMID- 16704294 TI - Recommendations for improving the home office project licence abstracts. AB - The Freedom of Information Act 2000 prompted the Government to evaluate how public understanding about the use of animals in scientific research can be improved and how animal experimentation is regulated within the UK. This resulted in the publication of anonymous project licence abstracts. Here, the abstracts published between December 2004 and October 2005 have been evaluated, by using a scoring system which takes into account their content and format, in order to assess whether they provide information that will facilitate the increased implementation of reduction, refinement and replacement strategies. The potential impact of freely-available Home Office project licence abstracts on public attitudes to animal testing and on the quality of animal-based research, is also discussed. Although some abstracts scored well, in general, the abstracts were found to be severely lacking in detail about the welfare of experimental animals. Thus, the abstracts tend to present a distorted picture of animal-based research. Many abstracts lack the details which would reveal how the licensing criteria had been met. It is concluded that, in their current form, the project abstracts are not sufficiently informative. A number of recommendations are made, which address this problem. PMID- 16704295 TI - A response to reviews by Russell, Festing and Patel, Hendriksen, and Thomas on the Nuffield Council on Bioethics Report, The ethics of research involving animals. PMID- 16704296 TI - Nicotinamide reduces high secretion of IFN-gamma in high-risk relatives even though it does not prevent type 1 diabetes. AB - Type 1 diabetes (T1D) is an autoimmune disease suggested to be of a T helper (Th)1-like origin. This study aimed to investigate the Th1-like and Th2-like profile in high-risk individuals during the prediabetic phase and the immunologic effect of treatment with nicotinamide. High-risk first-degree relatives of T1D patients participating in the European Nicotinamide Diabetes Intervention Trial (ENDIT) were treated with either nicotinamide or placebo. Peripheral blood mononuclear cells (PBMC) were obtained during the prediabetic phase and close to the onset of manifest T1D and from nondiabetic high-risk individuals. Using the sensitive enzyme-linked immunospot (ELISPOT) technique to distinguish Th1-like from Th2-like lymphocytes, secretion of interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) was analyzed from PBMCs spontaneously and after in vitro stimulation with the diabetes-associated autoantigens, glutamic acid decarboxylase 65 (GAD65, protein and peptide, aa 247-279), recombinant tyrosine phosphatase (IA-2), and heat shock protein (HSP, aa 437-460). High-risk individuals showed high spontaneous as well as autoantigen-induced IFN-gamma secretion. Secretion of IFN-gamma and the IFN-gamma/IL-4 ratio, induced by autoantigens, decreased in individuals developing T1D (p < 0.05), whereas nondiabetic individuals showed an increased IL-4 response (p < 0.05). Thus, a Th1 dominated cytokine profile observed in high-risk individuals inclined toward a diagnosis of diabetes. Nicotinamide caused decreased spontaneous (p = 0.05) and in vitro autoantigen-induced IFN-gamma secretion (p < 0.05) and may play a role in immune regulation, even though it has not been shown to prevent T1D. PMID- 16704297 TI - Stimulus-dependent and domain-dependent cell death acceleration by an IFN inducible protein, human MxA. AB - Human MxA is an interferon- alpha / beta (IFN-alpha/beta)-inducible protein that inhibits multiplication of influenza viruses and other RNA viruses. We reported that MxA accelerates cell death induced by apoptotic stimuli as well as influenza viral infection. However, the mechanism of MxA-mediated enhancement of cell death is not well understood. Here, we demonstrated that the cell death promotion activity of MxA was caspase dependent when cell death was induced by UV irradiation or cycloheximide (CHX). In contrast, in the case of cell death after influenza viral infection, MxA promoted both caspase-dependent and caspase independent cell death. The C-terminal region of MxA containing the oligomerization domain was found to be responsible for promotion of the cell death induced by CHX. In the case of cell death after influenza viral infection, both C-terminal and N-terminal regions were shown to be involved in cell death promotion, although the GTP-binding and GTP-hydrolysis activity dependent on a tripartite GTP-binding motif in the N-terminal region was not required for the cell death promotion activity of MxA. These results suggest that MxA accelerates cell death induced by influenza viral infection through at least two distinct pathways. PMID- 16704299 TI - Facilitation of survival in a rat fulminant hepatic failure model by combination therapy using recombinant G-CSF and tacrolimus. AB - The mortality rate of fulminant hepatic failure (FHF) is high because of retarded liver regeneration. Recombinant human granulocyte colony-stimulating factor (rHuG CSF) and tacrolimus are known to be immunosuppressive and supportive to liver regeneration. We investigated the effects of their combination therapy in a rat FHF model with a 68% partial hepatectomy and 24% liver necrosis. All rats without drug pretreatment died within 55 h. The median time was prolonged from 37 to 52 h by rHuG-CSF (250 microg/kg/day s.c. on days -5 to 0) and to 46 h by tacrolimus (0.5 mg/kg/day i.m. on days -2 to 0). Notably, the combination therapy facilitated DNA biosynthesis and survival prolongation, with a median of 77 h. The interferon-gamma (IFN-gamma) protein levels and natural killer cell (NK) activity in the liver were low at 12 h, and no further inhibition was detected by any treatment. Tacrolimus significantly upregulated the mRNA levels of insulin receptors and transforming growth factor-alpha (TGF-alpha), whereas rHuG-CSF did not. Regarding tissue remodeling-related factors, rHuG-CSF upregulated mRNA levels of vascular endothelial growth factor (VEGF) and matrix metalloproteinase- 9 (MMP-9), whereas tacrolimus did not. The combination treatment upregulated protein levels of both insulin receptors and VEGF. These results suggest that tacrolimus improves the hepatocyte replication and rHuG-CSF contributes to tissue reconstitution, and this combination therapy directly facilitates liver regeneration in the FHF model. PMID- 16704298 TI - OAS and PKR are not required for the antiviral effect of Ad:IFN-gamma against acute HSV-1 in primary trigeminal ganglia cultures. AB - Three interferon-gamma (IFN-gamma)-induced antiviral pathways have been reported. Involved antiviral proteins include: Mx, RNase L/2',5'-OAS, and protein kinase R (PKR). Involvement of OAS and PKR in IFN-gamma-induced anti-herpes simplex virus 1 (HSV-1) pathways has not been reported previously, but IFN-gamma induces OAS and PKR when other viruses invade the nervous system. The aim of the current study was to determine if the absence of intact OAS and PKR antiviral pathways affects the antiviral activity of IFN-gamma during acute HSV-1 infection within the trigeminal ganglia (TG). To investigate this, primary TG cultures were established using TGs removed from C57BL/6 (wild-type), RNase L knockout, and RNase L/PKR double knockout mice. Each dissociated TG was transduced with an adenoviral vector containing an IFN-gamma transgene or vector alone. Viral titers after HSV-1 infection of primary TG cell cultures were determined. Significant differences in viral titer for Ad:Null-transduced vs. Ad:IFN-gamma-tranduced TG were found in each genotype. However, the effectiveness of Ad:IFN-gamma was not reduced in the absence of both OAS and PKR pathways or OAS alone. Recombinant IFN gamma also exhibited anti-HSV-1 activity. The effectiveness of the IFN-gamma transgene was lost in primary TG cells from IFN-gamma receptor knockout mice. The data suggest that novel anti-HSV-1 mechanisms are induced by IFN-gamma. PMID- 16704301 TI - High IFN-alpha responses associated with depletion of lymphocytes and natural IFN producing cells during classical swine fever. AB - During the acute phase of the viral hemorrhagic disease, classical swine fever (CSF), a severe hematologic depletion in primary lymphoid organs and depletion of peripheral blood T and B lymphocytes are observed. The onset of these pathologic events is before viremia and independent of leukocyte infection, indicating a host-mediated effect possibly through a cytokine storm. Here, we show that high serum levels of interferon- alpha (IFN-alpha) were found during this phase of CSF, detectable as early as 2 days postinfection and reaching maximum levels 3-5 days postinfection (250-1300 U/mL). This IFN-alpha response was related to the virulence of the viral strain used, with avirulent virus not inducing any detectable serum IFN-alpha. A progressive depletion of natural IFN-producing cells/plasmacytoid dendritic cells (pDC), the likely in vivo source of IFN-alpha, was also induced by the viral infection. An important finding was that the onset of severe lymphopenia was concomitant with the IFN-alpha responses, and all animals with serum IFN-alpha had depleted B and T lymphocytes. A statistically significant correlation between lymphocyte depletion and serum IFN-alpha indicates a relationship between the two events, which is supported by the known hematologic effects of high IFN-alpha doses in vivo. PMID- 16704300 TI - Cell to cell contact through ICAM-1-LFA-1 and TNF-alpha synergistically contributes to GM-CSF and subsequent cytokine synthesis in DBA/2 mice induced by 1,3-beta-D-Glucan SCG. AB - SCG is a major 6-branched 1,3-beta-D-glucan in Sparassis crispa Fr. showing antitumor activity. We recently found that the splenocytes from naive DBA/1 and DBA/2 mice are potently induced by SCG to produce interferon- gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), granulocyte-macrophage colony stimulating factor (GM-CSF), and interleukin-12p70 (IL-12p70), and that GM-CSF plays a key biologic role among these cytokines. In this study, we investigated the contribution of cell-cell contact and soluble factors to cytokine induction by SCG in DBA/2 mice. Cell-cell contact involving intercellular adhesion molecule 1 (ICAM-1) and lymphocyte function-associated antigen-1 (LFA-1) was an essential step for the induction of GM-CSF and IFN-gamma by SCG but not for the induction of TNF-alpha or IL-12p70 by SCG. SCG directly induced adherent splenocytes to produce TNF-alpha and IL-12p70. GM-CSF was required for the induction of TNF alpha by SCG, and in turn, TNF-alpha enhanced the release of GM-CSF and thereby augmented the induction of IL-12p70 and IFN-gamma by SCG. Neutralization of IL-12 significantly inhibited the induction of IFN-gamma by SCG. We concluded that induction of GM-CSF production by SCG was mediated through ICAM-1 and LFA-1 interaction, GM-CSF subsequently contributed to further cytokine induction by SCG, and reciprocal actions of the cytokines were essential for enhancement of the overall response to SCG in DBA/2 mice. PMID- 16704302 TI - In vitro effects of recombinant zebrafish IFN on spring viremia of carp virus and infectious hematopoietic necrosis virus. AB - In order to evaluate the effects of fish recombinant interferon (rIFN) on fish pathogenic rhabdoviruses, the zebrafish (Danio rerio) IFN (DreIFN) allele B gene was cloned and expressed in Escherichia coli. In addition, the effects of recombinant DreIFN (rDreIFN) on spring viremia of carp virus (SVCV), infectious hematopoietic necrosis virus (IHNV), and vesicular stomatitis virus (VSV) were surveyed in fish and chicken cells. The mature peptide of DreIFN allele B gene encodes 163 amino acids. Residues 3 and 98 are a pair of cysteines that likely form an intrachain disulfide bridge. rDreIFN protein was detected as a band at 21.6 kDa by SDS-PAGE. The purified rDreIFN has anti-SVCV and anti-IHNV activity of 3 x 10(4) U/mg-10(7) U/mg. The results indicate that rDreIFN has higher activity against SVCV and IHNV on epithelioma papulosum cyprinid (EPC) than on grass carp (C. idellus) ovary (CO) cell lines and no activity against VSV on chick embryo fibroblasts (CEF). PMID- 16704304 TI - Functional analysis of nuclear pore complex protein Nup62/p62 using monoclonal antibodies. AB - The nuclear pore complex (NPC) is an enormous structure embedded in the double membrane of the nuclear envelope that acts as a passageway for nucleocytoplasmic transport. The vertebrate NPC is comprised of about 30 unique proteins. Nup62/p62, a major component of the NPC, has been reported to interact directly with several nuclear transport factors, including importin-beta and NTF2. However, it has not been shown how the interaction of Nup62/p62 with transport factors is involved in nucleocytoplasmic transport. The present study reports on the preparation of monoclonal antibodies (MAbs) directed against human Nup62/p62 and a functional analysis of Nup62/p62 using antibodies in living cells. Hybridomas producing the antibodies were produced by the hybridization of mouse myeloma cells with medial iliac lymph node cells from an immunized rat. These MAbs specifically recognized Nup62/p62 as evidenced by immunoblotting analysis using a nuclear membrane fraction. In the immunostaining using MAbs, a punctuate nuclear rim staining pattern was observed. Moreover, cytoplasmic injected-anti Nup62/p62 MAbs were rapidly targeted to the nuclear pore of cultured cells and some of them inhibited normal cell division, causing the formation of abnormal nuclei. The antibodies described in this study provide the means for immunochemical analyses of the NPC protein Nup62/p62 in mammalian cells, and represent useful molecular tools that should permit a better understanding of the biological roles and cellular dynamics of this protein in nucleocytoplasmic transport, cell division, and nuclear organization. PMID- 16704305 TI - Regulation of matrix metalloproteinase production and tumor cell invasion by four monoclonal antibodies against different epitopes of HAb18G/CD147 extracellular domain. AB - HAb18G/CD147, a membrane spanning molecule and highly expressed in hepatocellular carcinoma (HCC) cells, was shown to stimulate the production of matrix metalloproteinases (MMPs) in the interaction of tumor cells and fibroblasts. Studies on the EMMPRIN/CD147 showed that CD147 extracellular domain is involved in the induction of MMPs. To study the biological molecular function of HAb18G/CD147 extracellular domain (HAb18G/CD147-ED) on production of MMPs following mediated tumor cell invasion, we isolated four novel monoclonal anibodies (MAbs)-1B3, 3B3, HAb18Gedomab1, and HAb18Gedomab2-against HAb18G/CD147 ED by immunization of BALB/c mice with purified HAb18G/CD147-ED fragments, which were efficiently expressed in Escherichia coli. Gelatin zymography and Boyden chamber assays were used to identify the production of MMPs in the co-cultured human fibroblast and HCC cells, and to quantify the migrated cells in the presence of the generated MAbs. The results showed that two MAbs (1B3 and 3B3) inhibited [corrected] the secretion of MMP-2 and [corrected] the HCC cell invasion, whereas the other two MAbs (HAb18Gedomab1 and HAb18Gedomab2) had reverse function [corrected] FCM additive assay showed that four MAbs recognized different epitopes of HAb18G/CD147-ED. Taken together, the results suggest that various regions of HAb18G/CD147-ED participated in the regulation of MMP secretion. PMID- 16704306 TI - A rapid and efficient in vivo method for determining the biologic efficacy of monoclonal antibodies in animal models of cancer. AB - The selection of efficacious anti-tumor monoclonal antibodies (MAbs) for biological applications is a lengthy and labor-intensive process. In vitro characterization of one hybridoma fusion may reveal large numbers of tumor antigen-specific hybridomas. Very often, many of these tumor-specific antibodies need to be assessed in vivo using several different murine xenograft tumor prevention models to determine biological efficacy. The production and purification of sufficient quantities of many antigen-specific hybridomas is time consuming, and several months can pass between initial determination of MAb specificity and bioactivity. Moreover, many tumor-specific MAbs selected using in vitro binding studies have no in vivo anti-tumor efficacy. These studies describe an in vivo screening method either to eliminate non-efficacious MAbs or to rank order several tumor-specific MAbs in an expeditious manner. Proof-of-concept studies were conducted using two hybridomas secreting fully characterized neutralizing human anti-tumor MAbs (CNTO MAbs). Nu-/nu- mice were injected with CNTO MAb-secreting hybridoma cells in Matrigel cell matrix, followed by injection of target human tumor cells 4 days later (when circulating CNTO MAbs were detected in serum). Both the tumor take-rate and the mean tumor volumes were reduced significantly in mice treated with CNTO MAbsecreting hybridomas compared with mice treated with non-antibody-secreting cells. A panel of human antitumor antigen-specific MAbs with unknown biological efficacy was then evaluated by this method. The hybridomas exhibited a varied pattern of anti-tumor protection, indicating that some hybrids were secreting neutralizing anti-tumor MAbs, while others appeared to be less efficacious. These studies demonstrate a rapid, biologically relevant "yes/no" in vivo screening method for the evaluation of anti-tumor antigen MAbs. PMID- 16704307 TI - Monoclonal anti-thrombopoietin antibodies generated by genetic immunization. AB - Thrombopoietin (TPO) is a megakaryocyte growth and differentiation factor that is currently being investigated as a therapeutic for cancer patients undergoing myelosuppressive chemotherapy. We generated monoclonal antibodies (MAbs) specific for human thrombopoietin (hTPO) by genetic immunization using an hTPO expression plasmid and an adjuvant plasmid that encodes mouse granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4). All genetically immunized mice exhibited a high humoral immune response. Splenocytes from these mice were used to generate hybridomas. Two MAbs, designated 2B9A10 and 4C16B15 (of IgG1 and IgG3 isotypes, respectively), were subsequently selected and produced. They specifically recognized and precipitated recombinant hTPO produced by mammalian cells and were effective in sandwich enzyme-linked immunosorbent assays (ELISAs) for hTPO quantitation. Our results demonstrate that these MAbs should be useful for purification and quantitation of hTPO in clinical and laboratory settings. PMID- 16704308 TI - Preparation and characterization of a monoclonal antibody against human B lymphocyte stimulator. AB - B lymphocyte stimulator (BLyS) is a member of the tumor necrosis factor (TNF) family. It is required for B cell development. Deregulation of BLyS was involved in the pathogenesis of B cell-related autoimmune diseases and multiple myeloma. To prepare monoclonal antibodies (MAbs) against BLyS, cDNA encoding soluble BLyS (sBLyS) was first amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) with specific primers, and then inserted into a prokaryotic expression vector pET-30a. Right recombinant plasmid was expressed in Escherichia coli strain BL21(DE3), purified by nickel affinity chromatography. Isolated sBLys was used as an antigen to immunize mice. Splenocytes of one immunized mouse were fused with NS- 1. Hybridomas secreting antibodies against sBLyS were identified by ELISA. One positive clone was selected to produce antibody by injecting the hybridoma into the peritoneal cavity of mice. After collecting ascites, the antibody was purified by protein A affinity chromatography. Western blot and immunoflourescence demonstrated that the antibody could bind recombinant sBLyS and genuine membrane-bound BLyS (mBLyS) on U937. This MAb can be used as a detecting reagent to analyze the serum level of BLyS in patients with autoimmune diseases and the expression profile of mBLyS on multiple myeloma cells. PMID- 16704309 TI - Generation and characterization of monoclonal antibodies against FABP4. AB - Fatty acid binding protein 4 (FABP4) is a key mediator of intracellular transport and metabolism of fatty acids in adipose tissues. FABP4 binds fatty acids with high affinity and transports them to various compartments in the cell. When in complex with fatty acids, FABP4 interacts with and modulates the activity of two important regulators of metabolism: hormone-sensitive lipase and peroxisome proliferator-activated receptor gamma. Genetic studies in mice clearly indicated that deregulation of FABP4 function may lead to the development of severe diseases such as diabetes II type and atherosclerosis. In this study, we report the production and detailed characterization of monoclonal antibodies (MAbs) against FABP4. Recombinant glutathione S-transferase (GST)-FABP4 or His-FABP4 was expressed in bacteria, affinity purified, and used for immunization of mice, enzyme-linked immunosorbent assay (ELISA) screening, and characterization of selected clones. We have isolated two hybridoma clones that produced antibodies specific for recombinant and native FABP4, as shown by Western blotting and immunoprecipitation. The specificity of generated antibodies was further tested in a cell-based model of adipogenesis. In this analysis, the accumulation of FABP4 during NIH 3T3-L1 differentiation into adipocytes was detected by generated antibodies, which correlates well with previously published data. Taken together, we produced MAbs that will be useful for the scientific community working on fatty acid-binding proteins and lipid metabolism. PMID- 16704310 TI - Establishment and characterization of calcyclin binding protein (CacyBP) monoclonal antibody. AB - Anti-calcyclin binding protein (CacyBP) monoclonal antibodies (MAb) were produced using an in vitro immunization method. BALB/c mouse splenocytes were immunized with purified 6 x His-CacyBP fusion protein and fused with myeloma cells using polyethylene glycol (PEG) 4000. By selection using enzyme-linked immunosorbent assay (ELISA), three anti-CacyBP MAbs were obtained. The MAb BD1, whose isotype was IgG1, interacted with the fusion protein. Western blot and immunofluorescence microscopy showed that the MAb BD1 against CacyBP could recognize CacyBP protein derived from human gastric cancer cell lines in both native and denatured forms. This MAb would act as a useful tool for the detection of CacyBP protein in future studies. PMID- 16704311 TI - Preparation and identification of monoclonal antibodies against daintain. AB - Daintain is a 17-kDa polypeptide originally purified from porcine intestine. This polypeptide is associated with insulin secretion and inflammatory responses. Daintain is highly similar in amino acid sequence to allograft inflammatory factor-1 (AIF-1). Here we report the preparation and identification of monoclonal antibodies (MAbs) against daintain. To enhance its immunogenicity, daintain was coupled to carrier protein bovine serum albumin (BSA) by a two-step glutaraldehyde method. Using conventional procedures, we obtained four stable hybridoma cell lines that can produce and secret anti-daintain MAbs. We further analyzed their isotypes, titer, and affinity, and found that those MAbs belong to the G1 subclass with kappa light chains. The MAbs were capable of recognizing daintain as determined by Western blotting. The produced MAbs will be a useful tool for further investigation of daintain functions in organisms. PMID- 16704319 TI - 2005 Shneidman Award Address. Integratively assessing risk and protective factors for adolescent suicide. AB - This article briefly reviews key issues in adolescent suicide risk assessment and proposes that assessing risk and protective factors in combination has the best probability of informing the field's understanding of this complex problem. Several newer measures are described along with summaries of their psychometric properties. A recommended protocol utilizing 4 developmentally appropriate, valid, and reliable self-report measures is suggested as a parsimonious way of gathering information on the range of risk and protective factors. Finally, a novel approach of employing short-term longitudinal studies to test models of adolescent suicide attempts is described along with a rationale for not focusing research efforts on completed suicide. Implications of this approach for prevention efforts conclude the article. PMID- 16704320 TI - Negative bodily self in suicide attempters. AB - In the present study we investigated the relationship between suicidal behavior and aspects of bodily perception and parental care. Measures of bodily perception included measures of tactile sensitivity, body attitudes, and body experiences. Measures of parental care included parental bonding, negative and positive touch, and early maltreatment. One hundred and two adolescents (suicidal and nonsuicidal inpatients, and a control group) participated in the study. It was hypothesized that suicidal adolescents would (1) have higher tactile sensation thresholds, and more negative body attitudes and experiences; and (2) report less parental care, lower positive and higher negative parental touch, and higher parental maltreatment. It was also hypothesized that bodily sensitivity would mediate the relationship between suicidal tendencies and perceived negative early care. These hypotheses were confirmed. The role of a negative bodily self in suicidal behavior is proposed and discussed. PMID- 16704321 TI - Developmental differences in risk factors for suicide attempts between ninth and eleventh graders. AB - In order to identify differences in risk factors for suicide attempts throughout adolescence, this study utilized a school-based survey of ninth (n = 1,192) and eleventh graders (N = 1,055). Suicide attempts were associated with cigarette and alcohol use, family violence, and depression for ninth graders and with illicit drug use, school violence, and sexual abuse for eleventh graders, while having friends was protective for both groups. Additionally, having more than one risk factor imparted an exponential risk for suicide attempts (ninth > eleventh graders). The differences detected are consistent with developmental changes of adolescence and represent important information for identification of at-risk youth. PMID- 16704322 TI - Parents' and teachers' concordance with children's self-ratings of suicidality: findings from a high-risk sample. AB - This study examined concordance between adult and child reports of child suicidality using a sample of 1,046 8-year-old children at risk for, or having experienced, maltreatment. Concordance was low with both caregivers and teachers. For children reporting no suicidality, caregiver-child agreement was associated with few transitions in caregiver and low social withdrawal and aggression, and teacher-child agreement was associated with non-White ethnicity, good academic performance, and low thought problems. For children reporting suicidality, care giver-child agreement was associated with perceptions of child mental health needs, aggression, and somatic complaints, and teacher-child agreement was associated with thought problems. PMID- 16704323 TI - Strengthening the validity of population-based suicide rate comparisons: an illustration using U.S. military and civilian data. AB - The objectives of this study were to generate precise estimates of suicide rates in the military while controlling for factors contributing to rate variability such as demographic differences and classification bias, and to develop a simple methodology for the determination of statistically derived thresholds for detecting significant rate changes. Suicide rate estimates were calculated for the military population and each service branch over 11 years, directly standardized to the 2000 U.S. population. Military rates were highly comparable across branches and were approximately 20% lower than the civilian rate. Direct adjustment essentially controlled for the demographic confounds in this sample. Applying the Poisson-based method, we demonstrate that suicide rate fluctuations as large as 20-40% in any year may be attributed to random error. PMID- 16704324 TI - Epidemiology of nonfatal deliberate self-harm in the United States as described in three medical databases. AB - The absence of validated U.S. rates of nonfatal suicidal behavior places risk management and injury prevention programs at danger of being poorly informed and inadequately conceptualized. In this study we compare estimated rates of intentional self-harm from two ongoing surveys (National Electronic Injury Surveillance System-All Injury Program-NEISS-AIP; National Hospital Ambulatory Medical Care Survey-NHAMCS) to data from the Toxic Exposure Surveillance System. Results suggest that, for every 2002-2003 suicide, there were 12 (NEISSAIP) or 15 (NHAMCS) self-harm-related emergency department visits, and for every intentional self-poisoning death there were 33 intentional overdoses reported to poison control centers, of which two ultimately went untreated. PMID- 16704325 TI - Psychiatric hospitalization after deliberate self-poisoning. AB - The decision for psychiatric hospitalization after deliberate self-poisoning (DSP) is not well understood. This study, a longitudinal cohort study of 3,148 consecutive DSP patients found 920 (29.2%) subjects were referred for psychiatric hospitalization, 576 (18.3%) on involuntary basis. A logistic regression analysis showed increased risk for: age 25 or older, homelessness, unemployment, previous self-harm, psychiatric inpatient treatment within 12 months, earlier psychiatric inpatient treatment, suicidal ideation or plan, mood or psychotic disorders, and lower clinician experience; and lower risk for being married/defacto, and after hours presentation. Recommendation for psychiatric hospitalization was based on complex decision making. These findings have implications for clinical practice guidelines, service costs, and service organization. PMID- 16704326 TI - Clinical diagnostic and sociocultural dimensions of deliberate self-harm in Mumbai, India. AB - Patients' accounts complement psychiatric assessment of deliberate self-harm (DSH). In this study we examined psychiatric disorders, and sociocultural and cross-cultural features of DSH. SCID diagnostic interviews and a locally adapted EMIC interview were used to study 196 patients after DSH at a general hospital in Mumbai, India. Major depression was the most common diagnosis (38.8%), followed by substance use disorders (16.8%), but 44.4% of patients did not meet criteria for an enduring Axis-I disorder (no diagnosis, V-code, or adjustment disorder). Psychache arising from patient-identified sociocultural contexts and stressors complements, but does not necessarily fulfill, criteria for explanatory psychiatric disorders. PMID- 16704327 TI - Suicide risk at juvenile justice intake. AB - Many risks for suicidal behavior, identified in population samples, are elevated in justice youth. We examined whether risks operate similarly in a justice sample. We measured suicidal behavior and disorder on the Diagnostic Interview Schedule for Children in 991 randomly selected youths, and examined associations between demographic, offense, and disorder characteristics and past attempts. Recent attempts were more common in girls, in those with depression or substance disorder, and in violent offenders. While more girls reported recent attempts regardless of depression, depressed boys' attempt risk was as high as girls'. Depression contributed more to attempt history than did substance disorder. PMID- 16704328 TI - Stressful segregation housing and psychosocial vulnerability in prison suicide ideators. AB - Psychosocially vulnerable prisoners under stressful conditions of confinement are ill prepared to cope and at risk for developing suicide intention. The present study examined the relationships of depression, hopelessness, reasons for living, mental health problem history, suicide attempt lethality history, and stressful segregation housing with suicide ideation in 134 prisoners. Prisoners housed in segregation were found to have significantly higher levels of depression and suicide ideation than prisoners in general population. A hierarchical regression model of suicide ideation found significant interactions between mental health problem history, suicide attempt lethality history, and hopelessness with anticipated segregation stress, independent of depressed mood. Results of the study are discussed in terms of the stress-vulnerability model, various methodological limitations, and future research. PMID- 16704329 TI - A guide to the genetics of psychiatric disease. AB - The road to scientific discovery begins with an awareness of what is unknown. Research in science can in some ways be like putting together the pieces of a puzzle without having the benefit of the box-top picture of the completed puzzle. The "picture" in science is an understanding of how nature works in a particular instance, and it takes many separate pieces of the "puzzle" to put this understanding together. These pieces are always of different kinds of data, often obtained using different approaches and techniques. The challenge of the researcher is to picture or hypothesize each of the missing pieces before actually having them in hand, so they can be sought and tested in the laboratory. This "picturing" is actually having a clear idea of what you don't know: having a clear image of the "shape" of the missing piece. This is easy when the puzzle surrounding the missing piece is already in hand, but more difficult with less of it constrained by what is already known. In putting paper puzzles together, the shape of the pieces is not the only limitation that needs to be satisfied. There is also the picture to satisfy, that is, the picture usually has to make sense. In science these constraints can be manifold, and usually the quality of the research is judged by the number of ways a piece of data integrates into and brings together the rest of the puzzle. The multidimensionality of scientific questions makes it virtually essential that as many different pieces of the puzzle as possible be obtained. The more that is not known about the puzzle, the more pieces you need. Thus it is with the genetics of psychiatric diseases. In this guide, we will explore as many of the domains of the genetic puzzle as we are aware of. We will learn a bit of the language of each and how they fit into the puzzle with at least one anecdote to serve as an example. Mapping unknown territory is always a process, but we hope this guide will increase the reader's awareness of what is unknown. PMID- 16704330 TI - Self-reproach and personal responsibility. AB - A confusion exists between the aims of psychotherapy of diminishing self reproach, on the one hand, and increasing the experience of personal responsibility, on the other. In order to clarify this problem a distinction is made between moral responsibility, central to self-reproach, and psychological responsibility or agency. Self-reproach is shown to be inimical to the experience of psychological responsibility, with reference to psychotherapy of a case of severe obsessive self-reproach. PMID- 16704331 TI - Care: giving, receiving and meaning in the context of mental illness. AB - Understanding the dynamics of caregiving is essential to providing effective support to individuals and families living with serious mental illnesses. Yet, research in this area has typically explored caregiving without consulting mentally ill people. This paper adds this overlooked viewpoint by exploring the experience of care relationships by people diagnosed with schizophrenia. A secondary analysis of interview data from 21 individuals reveals that relevant dimensions of care relationships include: providers of care; types of care received; self-care; contested practices in care; negotiating practices; recipients of care; and types of care provided to other people. Addressing care within this broader conceptualization can contribute to developing interventions for individuals and families that more fully recognize the potential for people with mental illnesses to be active participants in care relationships. PMID- 16704332 TI - Untangling social function and social cognition: a review of concepts and measurement. AB - Over the past few decades, there has been increasing interest in the study of social impairment in schizophrenia. However, the concept of social functioning has been poorly defined in the literature. This article highlights the global and multi-factorial nature of social functioning and reviews the theoretical determinants of social dysfunction in schizophrenia. Emphasis is placed on outlining the social cognitive deficits that may occur. The study of social cognition appears particularly promising in elucidating our understanding of the development of social impairment in schizophrenia and has the potential to improve current psychosocial interventions. However, continued advances depend upon the existence of reliable and well-validated measures of social functioning and social cognition. A selection of measures are reviewed in this article in an attempt to highlight the importance of assessing multiple aspects of social functioning in schizophrenia and to assist researchers in the selection of appropriate measures. Future efforts should be directed towards the continued validation of social functioning and social cognitive measures and their adaptation for use in at-risk and early psychosis populations. PMID- 16704333 TI - Spinal cord injury, posttraumatic stress, and locus of control among the elderly: a comparison with young and middle-aged patients. AB - This study aimed to investigate the extent to which spinal cord injury posttraumatic stress (SCI PTSD) responses and the use of the external or internal health locus of control might vary according to age. Sixty-two patients with SCI were recruited for the study and divided into young (n = 23), middle-aged (n = 25) and elderly (n = 14) groups. They were assessed using the Posttraumatic Stress Disorder Checklist (PCL), the General Health Questionnaire-28 (GHQ-28), and the Multidimensional Health Locus of Control (MHLC). The results showed no significant differences between the three age groups in terms of PTSD symptoms. The elderly patients were significantly more socially dysfunctional than the other patients. The young patients believed in chance locus of control (CHLC) significantly more than the middle-aged and elderly patients. Correlation results revealed no significant relationship between PTSD symptoms and type of locus of control for the middle-aged patients. Otherwise, for both the young and elderly patients, internal health locus of control (IHLC) was negatively correlated with the avoidance symptom. For the elderly patients, powerful other locus of control (POLC) was positively correlated with the avoidance symptom. Both young and middle-aged patients revealed significant positive correlations between POLC, CHLC, and general health problems. For the elderly patients, POLC was positively correlated with social dysfunction and depression. SCI-PTSD responses did not differ according to age; however, the use of health locus of control differed depending on whether patients were younger or older. PMID- 16704334 TI - Enactment in schizophrenia: capacity for dialogue and the experience of the inability to commit to action. AB - While research has steadily begun to explore thoughts and beliefs linked with helplessness and despair in schizophrenia, it is less clearly understood how to account phenomenologically for the related experience of being unable to commit to action in the midst of grave discomfort. To explore this issue, the current paper presents an analysis of the experience of volitional paralysis of two persons over the course of long-term integrative psychotherapy. In particular, we explore the experience of the inability to commit to action and the consequences of the gradual recovery of a sense that one is capable of action. Results suggest that in both cases inaction was tethered to a sense of self as insufficiently centered to survive action. In particular, we suggest both men appeared initially unable to commit to action because such a commitment threatened them with forces both felt would undo the tenuous conversations that comprised their identities. Finally, as a sense of self as capable of action emerged, both men began to experience themselves as relatively more complex human beings and to sustain more complex conversation within themselves and between themselves and others. Implications for psychotherapy and rehabilitation are discussed. PMID- 16704336 TI - Mitochondrial fusion and fission in mammals. AB - Eukaryotic cells maintain the overall shape of their mitochondria by balancing the opposing processes of mitochondrial fusion and fission. Unbalanced fission leads to mitochondrial fragmentation, and unbalanced fusion leads to mitochondrial elongation. Moreover, these processes control not only the shape but also the function of mitochondria. Mitochondrial dynamics allows mitochondria to interact with each other; without such dynamics, the mitochondrial population consists of autonomous organelles that have impaired function. Key components of the mitochondrial fusion and fission machinery have been identified, allowing initial dissection of their mechanisms of action. These components play important roles in mitochondrial function and development as well as programmed cell death. Disruption of the fusion machinery leads to neurodegenerative disease. PMID- 16704337 TI - From nuclear transfer to nuclear reprogramming: the reversal of cell differentiation. AB - This is a personal historical account of events leading from the earliest success in vertebrate nuclear transfer to the current hope that nuclear reprogramming may facilitate cell replacement therapy. Early morphological evidence in Amphibia for the toti- or multipotentiality of some nuclei from differentiated cells first established the principle of the conservation of the genome during cell differentiation. Molecular markers show that many somatic cell nuclei are reprogrammed to an embryonic pattern of gene expression soon after nuclear transplantation to eggs. The germinal vesicles of oocytes in first meiotic prophase have a direct reprogramming activity on mammalian as well as amphibian nuclei and offer a route to identify nuclear reprogramming molecules. Amphibian eggs and oocytes have a truly remarkable ability to transcribe genes as DNA or nuclei, to translate mRNA, and to modify or localize proteins injected into them. The development of nuclear transplant embryos depends on the ability of cells to interpret small concentration changes of signal factors in the community effect and in morphogen gradients. Many difficulties in a career can be overcome by analyzing in increasing depth the same fundamentally interesting and important problem. PMID- 16704338 TI - How does voltage open an ion channel? AB - Neurons transmit information through electrical signals generated by voltage gated ion channels. These channels consist of a large superfamily of proteins that form channels selective for potassium, sodium, or calcium ions. In this review we focus on the molecular mechanisms by which these channels convert changes in membrane voltage into the opening and closing of "gates" that turn ion conductance on and off. An explosion of new studies in the last year, including the first X-ray crystal structure of a mammalian voltage-gated potassium channel, has led to radically different interpretations of the structure and molecular motion of the voltage sensor. The interpretations are as distinct as the techniques employed for the studies: crystallography, fluorescence, accessibility analysis, and electrophysiology. We discuss the likely causes of the discrepant results in an attempt to identify the missing information that will help resolve the controversy and reveal the mechanism by which a voltage sensor controls the channel's gates. PMID- 16704340 TI - Arsenic and selenium in microbial metabolism. AB - Arsenic and selenium are readily metabolized by prokaryotes, participating in a full range of metabolic functions including assimilation, methylation, detoxification, and anaerobic respiration. Arsenic speciation and mobility is affected by microbes through oxidation/reduction reactions as part of resistance and respiratory processes. A robust arsenic cycle has been demonstrated in diverse environments. Respiratory arsenate reductases, arsenic methyltransferases, and new components in arsenic resistance have been recently described. The requirement for selenium stems primarily from its incorporation into selenocysteine and its function in selenoenzymes. Selenium oxyanions can serve as an electron acceptor in anaerobic respiration, forming distinct nanoparticles of elemental selenium that may be enriched in (76)Se. The biogenesis of selenoproteins has been elucidated, and selenium methyltransferases and a respiratory selenate reductase have also been described. This review highlights recent advances in ecology, biochemistry, and molecular biology and provides a prelude to the impact of genomics studies. PMID- 16704339 TI - Curli biogenesis and function. AB - Curli are the major proteinaceous component of a complex extracellular matrix produced by many Enterobacteriaceae. Curli were first discovered in the late 1980s on Escherichia coli strains that caused bovine mastitis, and have since been implicated in many physiological and pathogenic processes of E. coli and Salmonella spp. Curli fibers are involved in adhesion to surfaces, cell aggregation, and biofilm formation. Curli also mediate host cell adhesion and invasion, and they are potent inducers of the host inflammatory response. The structure and biogenesis of curli are unique among bacterial fibers that have been described to date. Structurally and biochemically, curli belong to a growing class of fibers known as amyloids. Amyloid fiber formation is responsible for several human diseases including Alzheimer's, Huntington's, and prion diseases, although the process of in vivo amyloid formation is not well understood. Curli provide a unique system to study macromolecular assembly in bacteria and in vivo amyloid fiber formation. Here, we review curli biogenesis, regulation, role in biofilm formation, and role in pathogenesis. PMID- 16704341 TI - Manganese transport and the role of manganese in virulence. AB - Two areas of research have recently converged to highlight important roles for Mn(2+) in pathogenesis: the recognition that both bacterial Nramp homologs and members of LraI family of proteins are Mn(2+) transporters. Their mutation is associated with decreased virulence of various bacterial species. Thus, Mn(2+) appears to be essential for bacterial virulence. This review describes what is currently known about Mn(2+) transport in prokaryotes and how prokaryotic Mn(2+) transport is regulated. Some of the phenotypes that arise when microorganisms lack Mn(2+) are then discussed, with an emphasis on those phenotypes involving pathogenesis. The concluding section describes possible enzymatic roles for Mn(2+) that might help explain why Mn(2+) is necessary for virulence. PMID- 16704342 TI - The structural and functional role of RNA in icosahedral virus assembly. AB - Despite tremendous advances in high-resolution structure determination of virus particles, the organization of encapsidated genomes and their role during assembly are poorly understood. This article summarizes recent insights from structural, biochemical, and genetic analyses of icosahedral viruses that contain single-stranded, positive-sense RNA genomes. X-ray crystallography of several viruses in this category has provided tantalizing glimpses of portions of the packaged nucleic acid, contributing crucial information on how the genome might be folded within the virion. This information combined with theoretical considerations and data from molecular approaches suggests mechanisms by which coat proteins interact with genomic RNA to shape it into a conformation that is compatible with the geometry of the virion. It appears that RNA, in addition to its function as a repository for genetic information, plays an important structural role during assembly and can on occasion override the ability of the coat protein to form a particle with defined icosahedral symmetry. PMID- 16704344 TI - Uranium reduction. AB - The dramatic decrease in solubility accompanying the reduction of U(VI) to U(IV), producing the insoluble mineral uraninite, has been viewed as a potential mechanism for sequestration of environmental uranium contamination. In the past 15 years, it has been firmly established that a variety of bacteria exhibit this reductive capacity. To obtain an understanding of the microbial metal metabolism, to develop a practical approach for the acceleration of in situ bioreduction, and to predict the long-term fate of environmental uranium, several aspects of the microbial process have been experimentally explored. This review briefly addresses the research to identify specific uranium reductases and their cellular location, competition between uranium and other electron acceptors, attempts to stimulate in situ reduction, and mechanisms of reoxidation of reduced uranium minerals. PMID- 16704343 TI - Francisella tularensis: taxonomy, genetics, and Immunopathogenesis of a potential agent of biowarfare. AB - Tularemia is a zoonosis of humans caused by infection with the facultative intracellular bacterium Francisella tularensis. Interest in F. tularensis has increased markedly in the past few years because of its potential use as an agent of bioterrorism. Five subspecies of this organism are found in the Northern hemisphere, but only F. tularensis subsp. tularensis and subsp. holarctica cause disease in humans. This review summarizes what is known about the pathogenesis of tularemia with a focus on bacterial surface components such as lipopolysaccharide and capsule as well as information obtained from the F. tularensis subsp. tularensis SCHU S4 genome. In particular, the mechanisms of action of recently identified virulence factors are discussed in the context of bacterial replication in macrophages and manipulation of the host inflammatory response. Throughout this report, shared and unique features of F. tularensis subsp. tularensis, subsp. holarctica, and subsp. novicida are discussed. PMID- 16704345 TI - Radical enzymes in anaerobes. AB - This review describes enzymes that contain radicals and/or catalyze reactions with radical intermediates. Because radicals irreversibly react with dioxygen, most of these enzymes occur in anaerobic bacteria and archaea. Exceptions are the families of coenzyme B(12)- and S-adenosylmethionine (SAM)-dependent radical enzymes, of which some members also occur in aerobes. Especially oxygen-sensitive radical enzymes are the glycyl radical enzymes and 2-hydroxyacyl-CoA dehydratases. The latter are activated by an ATP-dependent one-electron transfer and act via a ketyl radical anion mechanism. Related enzymes are the ATP dependent benzoyl-CoA reductase and the ATP-independent 4-hydroxybenzoyl-CoA reductase. Ketyl radical anions may also be generated by one-electron oxidation as shown by the flavin-adenine-dinucleotide (FAD)- and [4Fe-4S]-containing 4 hydroxybutyryl-CoA dehydratase. Finally, two radical enzymes are discussed, pyruvate:ferredoxin oxidoreductase and methane-forming methyl-CoM reductase, which catalyze their main reaction in two-electron steps, but subsequent electron transfers proceed via radicals. PMID- 16704346 TI - The biology of the Cryptococcus neoformans species complex. AB - Cryptococcus neoformans is a major cause of fungal meningoencephalitis in immunocompromised patients. Despite recent advances in the genetics and molecular biology of C. neoformans, and improved techniques for molecular epidemiology, aspects of the ecology, population structure, and mode of reproduction of this environmental pathogen remain to be established. Application of recent insights into the life cycle of C. neoformans and its different ways of engaging in sexual reproduction under laboratory conditions has just begun to affect research on the ecology and epidemiology of this human pathogenic fungus. The melding of these disparate disciplines should yield rich dividends in our understanding of the evolution of microbial pathogens, providing insights relevant to diagnosis, treatment, and prevention. PMID- 16704347 TI - Maternal obesity, metabolism, and pregnancy outcomes. AB - About one third of all pregnant women in the United States are obese. Maternal obesity at conception alters gestational metabolic adjustments and affects placental, embryonic, and fetal growth and development. Neural tube defects and other developmental anomalies are more common in infants born to obese women; these defects have been linked to poor glycemic control. Preeclampsia, a gestational disorder occurring more frequently in obese women, appears to be due to a subclinical inflammatory state that impairs early placentation and development of its blood supply. Fetal growth and development during the last half of pregnancy depends on maternal metabolic adjustments dictated by placental hormones and the subsequent oxygen and nutrient supply. Maternal obesity affects these metabolic adjustments as well. Basal metabolic rates are significantly higher in obese women, and maternal fat gain is lower, possibly in response to altered leptin function. The usual increase in insulin resistance seen in late pregnancy is enhanced in obese mothers, causing marked postprandial increases in glucose, lipids, and amino acids and excessive fetal exposure to fuel sources, which in turn increases fetal size, fat stores, and risk for disease postnatally. Impaired glucose tolerance, gestational diabetes, and hyperlipidemia are more common among obese mothers. To date, little attention has been given to the role of diet among obese women in preventing these problems. However, studies of women with impaired glucose tolerance show that replacing refined carbohydrates and saturated fat with complex, low-glycemic carbohydrates and polyunsaturated fatty acids improves metabolic homeostasis and pregnancy outcomes. Thus, current dietary guidelines regarding the amount and type of carbohydrates and fat for nonpregnant women seem appropriate for pregnant women as well. PMID- 16704348 TI - Nutritional epidemiology and thyroid hormone metabolism. AB - Severe iodine deficiency was the main cause of endemic goiter and cretinism. Most of the previously iodine-deficient areas are now supplemented, mainly with iodized salt. The geographical distribution of severe endemic areas has been progressively reduced, and at present, approximately 200 million people living in remote places are still at risk of severe iodine deficiency. International public health programs should be focused first on reaching these populations, and second on auditing and monitoring the operational work of supplementation programs. This second point is essential to prevent iodine-induced hyperthyroidism or interruptions of iodine supplement distribution, which could be catastrophic for the fetus and the young infant. Echography brings a complementary tool to clinical assessment of goiter by palpation. Inductively coupled plasma-mass spectrometry brings at least a definitive gold standard for iodine measurement and thyroid hormone measurement. Thiocyanate overload has been clearly documented as a goitrogen in Central Africa, and when associated with selenium deficiency, it may be included as risk factor for endemic myxedematous cretinism. Variable exposure to different environmental risk factors is likely the explanation of the variable distribution of two types of endemic cretinism (neurological and myxedematous), and the clinical overlap of the pathogeny of both syndromes is more important than previously described. It is possible that Kashin-Beck osteoarthropathy is another evanescent endemic disease that will disappear with the correction of iodine deficiency. PMID- 16704350 TI - Variations on a gene: rare and common variants in ABCA1 and their impact on HDL cholesterol levels and atherosclerosis. AB - Cholesterol and its metabolites play a variety of essential roles in living systems. Virtually all animal cells require cholesterol, which they acquire through synthesis or uptake, but only the liver can degrade cholesterol. The ABCA1 gene product regulates the rate-controlling step in the removal of cellular cholesterol: the efflux of cellular cholesterol and phospholipids to an apolipoprotein acceptor. Mutations in ABCA1, as seen in Tangier disease, result in accumulation of cellular cholesterol, reduced plasma high-density lipoprotein cholesterol, and increased risk for coronary artery disease. To date, more than 100 coding variants have been identified in ABCA1, and these variants result in a broad spectrum of biochemical and clinical phenotypes. Here we review genetic variation in ABCA1 and its critical role in cholesterol metabolism and atherosclerosis in the general population. PMID- 16704351 TI - Immunonutrition in surgery and critical care. AB - The benefits of specialty supplemented enteral diets administered to critically ill and critically injured patients and those undergoing major surgical procedures have been documented in a number of randomized prospective studies. It is unclear which nutrient or combination of nutrients causes the beneficial effects, but there are significant reductions in infectious complications depending upon the patient populations studied. It is imperative that the data be interpreted in the context of individual patient risk since specialty formulas appear most beneficial in patients at risk of subsequent complications or in those with significant pre-existing malnutrition. Although controversy exists regarding the use of specialty supplemented enteral diets in critically ill patients, they have been administered safely with minimal risk of adverse outcome in malnourished patients and in the critically ill and critically injured. PMID- 16704352 TI - Control of energy homeostasis: role of enzymes and intermediates of fatty acid metabolism in the central nervous system. AB - The regulation of energy homeostasis is critical for normal physiology and survival. Energy flux must be rigorously monitored and adjusted to ensure that fuel intake and expenditure remain within acceptable limits. The central nervous system (CNS) is, in large part, responsible for conducting this energy-monitoring function and for integrating the numerous inputs. It has become evident that neurons of the CNS monitor and respond to levels of metabolic intermediates that reflect peripheral energy status. Intermediates in the fatty acid biosynthetic pathway have been implicated as hypothalamic signaling mediators that sense and respond to changes in circulating fuels. Genetic and pharmacologic manipulation of the enzymes of fatty acid metabolism have led to the hypothesis that neuronal metabolic intermediates affect neural outputs that modify both feeding behavior and energy expenditure. This review focuses on the regulatory roles of these enzymes and intermediates in the regulation of food intake and energy balance. PMID- 16704354 TI - Results of bariatric surgery. AB - Overweight and obesity are rapidly growing to epidemic proportions in the United States and globally. Since sustainable weight loss is only achieved by bariatric surgery, medicine has seen an explosion in the diversity and number of bariatric procedures performed over the past few years. Systematic studies of postoperative outcomes and investigations into the physiology and biology of weight loss provide a more comprehensive understanding of the sequelae of bariatric surgery. Adipose tissue is the predominant site of fat stores. Increasing obesity results in an overload of lipids within the body's natural storage sink (i.e., the adipocyte) followed by the necessary deposition of fat within ectopic sites such as muscle, liver, and pancreas. The resulting metabolic derangements are associated with insulin resistance, central obesity, and chronic inflammation as adipose tissue acts as an endocrine organ, producing and secreting a host of biologic mediators. Whereas there are conflicting data on the cardiovascular effects of peripheral, subcutaneous liposuction, malabsorptive bariatric procedures result almost universally in significant amelioration of insulin resistance, hypertension, dyslipidemia, and hepatic steatosis. Concomitant changes in adipocyte-derived hormones may provide mechanistic explanations to the observed improvements. PMID- 16704355 TI - Fitness of human enteric pathogens on plants and implications for food safety. AB - The continuous rise in the number of outbreaks of foodborne illness linked to fresh fruit and vegetables challenges the notion that enteric pathogens are defined mostly by their ability to colonize the intestinal habitat. This review describes the epidemiology of produce-associated outbreaks of foodborne disease and presents recently acquired knowledge about the behavior of enteric pathogens on plants, with an emphasis on Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes. The growth and survival of enteric pathogens on plants are discussed in the light of knowledge and concepts in plant microbial ecology, including epiphytic fitness, the physicochemical nature of plant surfaces, biofilm formation, and microbe-microbe and plant-microbe interactions. Information regarding the various stresses that affect the survival of enteric pathogens and the molecular events that underlie their interactions in the plant environment provides a good foundation for assessing their role in the infectious dose of the pathogens when contaminated fresh produce is the vehicle of illness. PMID- 16704357 TI - Comparative genomics reveals what makes an enterobacterial plant pathogen. AB - The bacterial family Enterobacteriaceae contains some of the most devastating human and animal pathogens, including Escherichia coli, Salmonella enterica and species of Yersinia and Shigella. These are among the best-studied of any organisms, yet there is much to be learned about the nature and evolution of interactions with their hosts and with the wider environment. Comparative and functional genomics have fundamentally improved our understanding of their modes of adaptation to different ecological niches and the genes that determine their pathogenicity. In addition to animal pathogens, Enterobacteriaceae include important plant pathogens, such as Erwinia carotovora subsp. atroseptica (Eca), the first plant-pathogenic enterobacterium to be sequenced. This review focuses on genomic comparisons between Eca and other enterobacteria, with particular emphasis on the differences that exemplify or explain the plant-associated lifestyle(s) of Eca. Horizontal gene transfer in Eca may directly have led to the acquisition of a number of determinants that mediate its interactions, pathogenic or otherwise, with plants, offering a glimpse into its evolutionary divergence from animal-pathogenic enterobacteria. PMID- 16704358 TI - The dawn of fungal pathogen genomics. AB - Recent advances in sequencing technologies have led to a remarkable increase in the number of sequenced fungal genomes. Several important plant pathogenic fungi are among those that have been sequenced or are being sequenced. Additional fungal pathogens are likely to be sequenced in the near future. Analysis of the available genomes has provided useful information about genes that may be important for plant infection and colonization. Genome features, such as repetitive sequences, telomeres, conserved syntenic blocks, and expansion of pathogenicity-related genes, are discussed in detail with Magnaporthe oryzae (M. grisea) and Fusarium graminearum as examples. Functional and comparative genomic studies in plant pathogenic fungi, although still in the early stages and limited to a few pathogens, have enormous potential to improve our understanding of the molecular mechanisms involved in host-pathogen interactions. Development of advanced genomics tools and infrastructure is critical for efficient utilization of the vast wealth of available genome sequence information and will form a solid foundation for systems biology studies of plant pathogenic fungi. PMID- 16704359 TI - A model plant pathogen from the kingdom Animalia: Heterodera glycines, the soybean cyst nematode. AB - The soybean cyst nematode, Heterodera glycines, adversely affects the production of soybean, Glycine max, in many areas of the world, particularly in the United States, where it is the most economically important soybean pathogen. Despite the availability of hundreds of H. glycines-resistant soybean cultivars, the nematode continues to be a major limiting factor in soybean production. The use of nonhost rotation and resistance are the primary means of reducing losses caused by the nematode, but each of these options has disadvantages. As a subject for study of nematode parasitism and virulence, H. glycines provides a useful model despite its obligately parasitic nature. Its obligately sexual reproduction and ready adaptation to resistant cultivars, formerly referred to as "race shift," presents an excellent opportunity for the study of virulence in nematodes. Recent advances in H. glycines genomics have helped identify putative nematode parasitism genes, which, in turn, will aid in the understanding of nematode pathogenicity and virulence and may provide new targets for engineering nematode resistance. PMID- 16704356 TI - Long-distance RNA-RNA interactions in plant virus gene expression and replication. AB - The vast majority of plant and animal viruses have RNA genomes. Viral gene expression and replication are controlled by cis-acting elements in the viral genome, which have been viewed conventionally as localized structures. However, recent research has altered this perception and provided compelling evidence for cooperative activity involving distantly positioned RNA elements. This chapter focuses on viral RNA elements that interact across hundreds or thousands of intervening nucleotides to control translation, genomic RNA synthesis, and subgenomic mRNA transcription. We discuss evidence supporting the existence and function of the interactions, and speculate on the regulatory roles that such long-distance interactions play in the virus life cycle. We emphasize viruses in the Tombusviridae and Luteoviridae families in which long-distance interactions are best characterized, but similar phenomena in other viruses are also discussed. Many more examples likely remain undiscovered. PMID- 16704373 TI - The histone acetyltransferases CBP/p300 are degraded in NIH 3T3 cells by activation of Ras signalling pathway. AB - The CBP [CREB (cAMP-response-element-binding protein)-binding protein]/p300 acetyltransferases function as transcriptional co-activators and play critical roles in cell differentiation and proliferation. Accumulating evidence shows that alterations of the CBP/p300 protein levels are linked to human tumours. In the present study, we show that the levels of the CBP/p300 co-activators are decreased dramatically by continuous PDGF (platelet-derived growth factor) and Ras signalling pathway activation in NIH 3T3 fibroblasts. This effect occurs by reducing the expression levels of the CBP/p300 genes. In addition, CBP and p300 are degraded by the 26 S proteasome pathway leading to an overall decrease in the levels of the CBP/p300 proteins. Furthermore, we provide evidence that Mdm2 (murine double minute 2), in the presence of active H-Ras or N-Ras, induces CBP/p300 degradation in NIH 3T3 cells. These findings support a novel mechanism for modulating other signalling transduction pathways that require these common co-activators. PMID- 16704375 TI - Nutrigenomics and nutrigenetics: the 'omics' revolution in nutritional science. AB - The execution of the Human Genome Project has brought forth a wealth of information about the structure of the genome, which can now be used to study how the interplay between our genes and factors from the environment such as nutrition relate to a state of health or disease. To enable such studies, novel technologies have been designed in particular to monitor the activity of multiple genes simultaneously at the level of the RNA by transcriptomics, or the level of the proteins by proteomics. In addition, genome information has boosted approaches to study the role of genetic variation to explain individual differences in responses to nutrition, underlying in part the susceptibility for nutrition-related disorders. These new areas of science referred to as 'nutrigenomics' and 'nutrigenetics' respectively, will increase our fundamental knowledge of the interaction between life processes and our diet or specific components thereof, which may in time lead to the development of novel functional foods to improve the health status of the general population, and to the personalized diet to prevent the onset of nutrition-related disorders in genetically predisposed individuals. PMID- 16704376 TI - Nuclear architecture in the light of gene expression and cell differentiation studies. AB - It is evident that primary DNA sequences, that define genomes, are responsible for genome functions. However, the functional properties of chromatin are additionally regulated by heritable modifications known as epigenetic factors and, therefore, genomes should be also considered with respect to their 'epigenomes'. Nucleosome remodelling, DNA methylation and histone modifications are the most prominent epigenetic changes that play fundamental roles in the chromatin-mediated control of gene expression. Another important nuclear feature with functional relevance is the organization of mammalian chromatin into distinct chromosome territories which are surrounded by the interchromatin compartment that is necessary for transport of regulatory molecules to the targeted DNA. The inner structure of the chromosome territories, as well as the arrangement of the chromosomes within the interphase nuclei, has been found to be non-randomly organized. Therefore, a specific nuclear arrangement can be observed in many cellular processes, such as differentiation and tumour cell transformation. PMID- 16704374 TI - Determination of the consensus DNA-binding sequence and a transcriptional activation domain for ESE-2. AB - The ESE (epithelium-specific Ets) subfamily of Ets transcription factors plays an important role in regulating gene expression in a variety of epithelial cell types. Although ESE proteins have been shown to bind to regulatory elements of some epithelial genes, the optimal DNA-binding sequence has not been experimentally ascertained for any member of the ESE subfamily of transcription factors. This has made the identification and validation of their targets difficult. We are studying ESE-2 (Elf5), which is highly expressed in epithelial cells of many tissues including skin keratinocytes. Here, we identify the preferred DNA-binding site of ESE-2 by performing CASTing (cyclic amplification and selection of targets) experiments. Our analysis shows that the optimal ESE-2 consensus motif consists of a GGA core and an AT-rich 5'- and 3'-flanking sequences. Mutational and competition experiments demonstrate that the flanking sequences that confer high DNA-binding affinity for ESE-2 show considerable differences from the known consensus DNA-binding sites of other Ets proteins, thus reinforcing the idea that the flanking sequences may impart recognition specificity for Ets proteins. In addition, we have identified a novel isoform of murine ESE-2, ESE-2L, that is generated by use of a hitherto unreported new exon and an alternate promoter. Interestingly, transient transfection assays with an optimal ESE-2 responsive reporter show that both ESE-2 and ESE-2L are weak transactivators. However, similar studies utilizing GAL4 chimaeras of ESE-2 demonstrate that while the DNA-binding ETS (E twenty-six) domain functions as a repressor, the PNT (pointed domain) of ESE-2 can act as a potent transcriptional activation domain. This novel transactivating property of PNT is also shared by ESE-3, another ESE family member. Identification of the ESE-2 consensus site and characterization of the transcriptional activation properties of ESE-2 shed new light on its potential as a regulator of target genes. PMID- 16704377 TI - Regulation of the actin cytoskeleton by PIP2 in cytokinesis. AB - Cytokinesis is a sequential process that occurs in three phases: assembly of the cytokinetic apparatus, furrow progression and fission (abscission) of the newly formed daughter cells. The ingression of the cleavage furrow is dependent on the constriction of an equatorial actomyosin ring in many cell types. Recent studies have demonstrated that this structure is highly dynamic and undergoes active polymerization and depolymerization throughout the furrowing process. Despite much progress in the identification of contractile ring components, little is known regarding the mechanism of its assembly and structural rearrangements. PIP2 (phosphatidylinositol 4,5-bisphosphate) is a critical regulator of actin dynamics and plays an essential role in cell motility and adhesion. Recent studies have indicated that an elevation of PIP2 at the cleavage furrow is a critical event for furrow stability. In this review we discuss the role of PIP2-mediated signalling in the structural maintenance of the contractile ring and furrow progression. In addition, we address the role of other phosphoinositides, PI(4)P (phosphatidylinositol 4-phosphate) and PIP3 (phosphatidylinositol 3,4,5 triphosphate) in these processes. PMID- 16704378 TI - Not the same everywhere. Patient-centered learning environments at nine medical schools. AB - BACKGROUND: Learning environments overtly or implicitly address patient-centered values and have been the focus of research for more than 40 years, often in studies about the "hidden curriculum." However, many of these studies occurred at single medical schools and used time-intensive ethnographic methods. This field of inquiry lacks survey methods and information about how learning environments differ across medical schools. OBJECTIVE: To examine patient-centered characteristics of learning environments at 9 U.S. medical schools. DESIGN: Cross sectional internet-based survey. PARTICIPANTS: Eight-hundred and twenty-three third- and fourth-year medical students in the classes of 2002 and 2003. MEASUREMENTS: We measured the patient-centeredness of learning environments with the Communication, Curriculum, and Culture (C3) Instrument, a 29-item validated measure that characterizes the degree to which a medical school's environment fosters patient-centered care. The C3 Instrument contains 3 content areas (role modeling, students' experiences, and support for students' patient-centered behaviors), and is designed to measure these areas independent of respondents' attitudes about patient-centered care. We also collected demographic and attitudinal information from respondents. RESULTS: The variability of C3 scores across schools in each of the 3 content areas of the instrument was striking and statistically significant (P values ranged from .001 to .004). In addition, the patterns of scores on the 3 content areas differed from school to school. CONCLUSIONS: The 9 schools demonstrated unique and different learning environments both in terms of magnitude and patterns of characteristics. Further multiinstitutional study of hidden curricula is needed to further establish the degree of variability that exists, and to assist educators in making informed choices about how to intervene at their own schools. PMID- 16704379 TI - Measuring outcomes of a one-minute preceptor faculty development workshop. AB - BACKGROUND: Measuring outcomes of faculty development programs is difficult and infrequently attempted beyond measuring participant satisfaction with the program. Few studies have validated evaluation tools to assess the effectiveness of faculty development programs, and learners have rarely participated in assessing improvement of faculty who participate in such programs. OBJECTIVE: To develop a questionnaire to measure the effectiveness of an enhanced 1-minute preceptor (OMP) faculty development workshop via faculty self-assessment and resident assessment of faculty, and to use the questionnaire to assess an OMP faculty development workshop. DESIGN AND MEASUREMENTS: We developed and tested a questionnaire to assess the 5 "microskills" of a OMP faculty development program, and performed faculty self-assessment and resident assessment using the questionnaire 6 to 18 months before and 6 to 18 months after our experiential skills improvement workshop. PARTICIPANTS: Sixty-eight internal medicine continuity clinic preceptors (44 control and 24 intervention faculty) at a university, a veteran's affairs hospital, and 2 community internal medicine training sites. RESULTS: Twenty-two participants (92%) completed pre- and postintervention questionnaires. Residents completed 94 preintervention questionnaires and 58 postintervention questionnaires on participant faculty. Faculty reported improvement in behavior following the intervention. Residents reported no significant improvements in faculty teaching behaviors following the intervention. CONCLUSION: We attempted to rigorously evaluate a faculty development program based on the OMP. Although the intervention did not show statistically significant changes in teaching behavior, we believe that this study is an important step in extending assessment of faculty development to include resident evaluation of participating faculty. PMID- 16704380 TI - Relationship between anxiety and standardized patient test performance in the medicine clerkship. AB - PURPOSE: Anxiety is thought to affect test performance. Studies have shown that students with low levels of test anxiety achieve higher scores on multiple choice question (MCQ) examinations than those with high anxiety levels. Female students have been shown to have higher test anxiety levels than male students. Standardized patient (SP) examinations are being used in medical schools and for licensing purposes. As SP exams are relatively new, there are few studies measuring anxiety levels for the SP test. The purpose of this study was to measure and compare medicine clerkship student SP versus MCQ examination anxiety levels and to determine if level affected test performance. METHODS: The Spielberger test attitude inventory was used to measure anxiety in 150 students rotating through the clerkship. Students completed questionnaires after the MCQ and SP examinations. Mean examination scores and anxiety levels were compared. Based on questionnaire scores, students were divided into 3 groups: low, moderate, and high anxiety. The MCQ and SP examination scores were analyzed to determine if male/female anxiety-level affected test performance. RESULTS: There were no meaningful anxiety-level differences between the SP and MCQ examinations. An inverse relationship between anxiety level and test scores was not identified. Female students had higher anxiety levels but sex differences did not influence examination performance. DISCUSSION: Medicine clerkship student test performance is not affected by anxiety level. Implications of the findings for incorporating stress management training in medical school curricula and suggestions for future research are discussed. PMID- 16704381 TI - Learning from mistakes. Factors that influence how students and residents learn from medical errors. AB - CONTEXT: Trainees are exposed to medical errors throughout medical school and residency. Little is known about what facilitates and limits learning from these experiences. OBJECTIVE: To identify major factors and areas of tension in trainees' learning from medical errors. DESIGN, SETTING, AND PARTICIPANTS: Structured telephone interviews with 59 trainees (medical students and residents) from 1 academic medical center. Five authors reviewed transcripts of audiotaped interviews using content analysis. RESULTS: Trainees were aware that medical errors occur from early in medical school. Many had an intense emotional response to the idea of committing errors in patient care. Students and residents noted variation and conflict in institutional recommendations and individual actions. Many expressed role confusion regarding whether and how to initiate discussion after errors occurred. Some noted the conflict between reporting errors to seniors who were responsible for their evaluation. Learners requested more open discussion of actual errors and faculty disclosure. No students or residents felt that they learned better from near misses than from actual errors, and many believed that they learned the most when harm was caused. CONCLUSIONS: Trainees are aware of medical errors, but remaining tensions may limit learning. Institutions can immediately address variability in faculty response and local culture by disseminating clear, accessible algorithms to guide behavior when errors occur. Educators should develop longitudinal curricula that integrate actual cases and faculty disclosure. Future multi-institutional work should focus on identified themes such as teaching and learning in emotionally charged situations, learning from errors and near misses and balance between individual and systems responsibility. PMID- 16704382 TI - A randomized trial of teaching clinical skills using virtual and live standardized patients. AB - BACKGROUND: We developed computer-based virtual patient (VP) cases to complement an interactive continuing medical education (CME) course that emphasizes skills practice using standardized patients (SP). Virtual patient simulations have the significant advantages of requiring fewer personnel and resources, being accessible at any time, and being highly standardized. Little is known about the educational effectiveness of these new resources. We conducted a randomized trial to assess the educational effectiveness of VPs and SPs in teaching clinical skills. OBJECTIVE: To determine the effectiveness of VP cases when compared with live SP cases in improving clinical skills and knowledge. DESIGN: Randomized trial. PARTICIPANTS: Fifty-five health care providers (registered nurses 45%, physicians 15%, other provider types 40%) who attended a CME program. INTERVENTIONS: Participants were randomized to receive either 4 live cases (n=32) or 2 live and 2 virtual cases (n=23). Other aspects of the course were identical for both groups. RESULTS: Participants in both groups were equivalent with respect to pre-post workshop improvement in comfort level (P=.66) and preparedness to respond (P=.61), to screen (P=.79), and to care (P=.055) for patients using the skills taught. There was no difference in subjective ratings of effectiveness of the VPs and SPs by participants who experienced both (P=.79). Improvement in diagnostic abilities were equivalent in groups who experienced cases either live or virtually. CONCLUSIONS: Improvements in performance and diagnostic ability were equivalent between the groups and participants rated VP and SP cases equally. Including well-designed VPs has a potentially powerful and efficient place in clinical skills training for practicing health care workers. PMID- 16704383 TI - The impact of the Stanford Faculty Development Program on ambulatory teaching behavior. AB - CONTEXT: Faculty development has received considerable investment of resources from medical institutions, though the impact of these efforts has been infrequently studied. OBJECTIVE: To measure the impact of the Stanford Faculty Development Program in Clinical Teaching on ambulatory teaching behavior. DESIGN: Pre-post. SETTING AND PARTICIPANTS: Eight internal medicine faculty participating in local faculty development. INTERVENTION: Participants received 7 2-hour sessions of faculty development. Each session included didactic, role-play, and videotaped performance evaluation. MAIN OUTCOME MEASURE: Before and after the intervention, faculty were video-taped during a case presentation from a standardized learner, who had been trained to portray 3 levels of learners: a third-year medical student, an intern, and a senior medical resident. Teacher and learner utterances (i.e, phrases) were blindly and randomly coded, using the Teacher Learner Interaction Analysis System, into categories that capture both the nature and intent of the utterances. We measured change in teaching behavior as detected through analysis of the coded utterances. RESULTS: Among the 48 videotaped encounters, there were a total of 7,119 utterances, with 3,203 (45%) by the teacher. Examining only the teacher, the total number of questions asked declined (714 vs 426, P=.02) with an increase in the proportion of higher-level, analytic questions (44% vs 55%, P<.0001). The quality of feedback also improved, with less "minimal" feedback (87% vs 76%, P<.0005) and more specific feedback (13% vs 22%) provided. CONCLUSIONS: Teaching behaviors improved after participation in this faculty development program, specifically in the quality of questions asked and feedback provided. PMID- 16704384 TI - What do attending physicians contribute in a house officer-based ambulatory continuity clinic? AB - OBJECTIVE: To study the educational contributions of attending physicians in an internal medicine house staff ambulatory clinic. DESIGN: Cross-sectional, self administered survey. SETTING: University-affiliated general internal medicine practice. PATIENTS/PARTICIPANTS: Internal medicine residents and attendings. MEASUREMENTS AND MAIN RESULTS: Attending and resident perceptions of whether attendings made contributions to teaching points, diagnosis (DX), therapy (RX), and health care maintenance (HCM) were assessed in 428 patient encounters. Resident assessments significantly exceeded attending self-assessments of contributions to teaching points (82% vs 74%, P=.001), DX (44% vs 34%, P=.001), RX (61% vs 55%, P=.02), and HCM (19% vs 15%, P=.04). Both residents and attendings perceived that contributions declined progressively with increasing resident year (P<.05). Primary care and categorical residents assessed attending contributions comparably. However, attendings perceived contributing more to RX and HCM for categorical residents than primary care (P<.05). Male and female residents assessed attending contributions comparably. However, attendings perceived contributing generally more to DX in male residents than female (P=.003). In 8% of encounters, either residents or attendings felt that patient evaluation by the attending was needed. In these encounters with personal patient evaluation by attendings, both residents and attendings felt that attendings made more contributions to DX (P=.001) and teaching points than in other encounters. CONCLUSIONS: Attending physicians consistently underestimate their perceived contributions to house officer ambulatory teaching. Their personal patient evaluation increases assistance with DX and teaching points. Given perceived declining contributions by training year, attendings may need to identify other teaching strategies for interactions with senior residents. PMID- 16704385 TI - Physician characteristics associated with proficiency in feedback skills. AB - BACKGROUND: Providing and eliciting high-quality feedback is valuable in medical education. Medical learners' attainment of clinical competence and professional growth can be facilitated by reliable feedback. This study's primary objective was to identify characteristics that are associated with physician teachers' proficiency with feedback. METHODS: A cohort of 363 physicians, who were either past participants of the Johns Hopkins Faculty Development Program or members of a comparison group, were surveyed by mail in July 2002. Survey questions focused on personal characteristics, professional characteristics, teaching activities, self-assessed teaching proficiencies and behaviors, and scholarly activity. The feedback scale, a composite feedback variable, was developed using factor analysis. Logistic regression models were then used to determine which faculty characteristics were independently associated with scoring highly on a dichotomized version of the feedback scale. RESULTS: Two hundred and ninety-nine physicians responded (82%) of whom 262 (88%) had taught medical learners in the prior 12 months. Factor analysis revealed that the 7 questions from the survey addressing feedback clustered together to form the "feedback scale" (Cronbach's alpha: 0.76). Six items, representing discrete faculty responses to survey questions, were independently associated with high feedback scores: (i) frequently attempting to detect and discuss the emotional responses of learners (odds ratio [OR]=4.6, 95% confidence interval [CI] 2.2 to 9.6), (ii) proficiency in handling conflict (OR=3.7, 95% CI 1.5 to 9.3), (iii) frequently asking learners what they desire from the teaching interaction (OR=3.5, 95% CI 1.7 to 7.2), (iv) having written down or reviewed professional goals in the prior year (OR=3.2, 95% CI 1.6 to 6.4), (v) frequently working with learners to establish mutually agreed upon goals, objectives, and ground rules (OR=2.2, 95% CI 1.1 to 4.7), and (vi) frequently letting learners figure things out themselves, even if they struggle (OR=2.1, 95% CI 1.1 to 3.9). CONCLUSIONS: Beyond providing training in specific feedback skills, programs that want to improve feedback performance among their faculty may wish to promote the teaching behaviors and proficiencies that are associated with high feedback scores identified in this study. PMID- 16704386 TI - Teaching internal medicine residents in the new era. Inpatient attending with duty-hour regulations. AB - BACKGROUND: Little is known about the impact of resident duty-hour regulations on the inpatient teaching experience. OBJECTIVE: Provide descriptive information on the effect of resident duty-hour regulations on attendings and the educational environment. DESIGN: Qualitative analysis of attending focus groups and e-mail survey of residents in Internal Medicine. PARTICIPANTS: Inpatient attending physicians at 2 academic centers and residents at the affiliated university-based Internal Medicine residency program in Portland, OR. RESULTS: Seventy-two percent of eligible attendings participated in 2 focus groups. Three themes were generated: increased clinical role, altered time management, and altered teaching. Attending physicians report performing more clinical work, teaching less, using more focused teaching methods, and experiencing an increased perception of intensity. Forty percent of eligible residents completed our e-mail survey. We organized residents data using the same 3 themes as attending physician data. Residents observed attending physicians performing increased clinical work, being more time aware, delivering more focused teaching, and having less time to teach. Participants noted changes in autonomy and professionalism. Strategies to enhance teaching effectiveness in the new environment were described. CONCLUSION: Duty-hour regulations have increased attending clinical responsibility and decreased teaching time in 1 residency program, leading to the perception of a more intense attending experience. Duty hour regulations encourage educators to determine what is critical to preserve in the educational experiences of learners and challenge us to reexamine autonomy and professionalism in training. PMID- 16704387 TI - Teaching about Substance Abuse with Objective Structured Clinical Exams. AB - BACKGROUND: Although residents commonly manage substance abuse disorders, optimal approaches to teaching these specialized interviewing and intervention skills are unknown. OBJECTIVE: We developed a Substance Abuse Objective Structured Clinical Exam (OSCE) to teach addiction medicine competencies using immediate feedback. In this study we evaluated OSCE performance, examined associations between performance and self-assessed interest and competence in substance abuse, and assessed learning during the OSCE. DESIGN: Five-station OSCE, including different substance abuse disorders and readiness to change stages, administered during postgraduate year-3 ambulatory rotations for 2 years. PARTICIPANTS: One hundred and thirty-one internal and family medicine residents. MEASUREMENTS: Faculty and standardized patients (SPs) assessed residents' general communication, assessment, management, and global skills using 4-point scales. Residents completed a pre-OSCE survey of experience, interest and competence in substance abuse, and a post-OSCE survey evaluating its educational value. Learning during the OSCE was also assessed by measuring performance improvement from the first to the final OSCE station. RESULTS: Residents performed better (P<.001) in general communication (mean+/-SD across stations=3.12+/-0.35) than assessment (2.65+/ 0.32) or management (2.58+/-0.44), and overall ratings were lowest in the contemplative alcohol abuse station (2.50+/-0.83). Performance was not associated with residents' self-assessed interest or competence. Perceived educational value of the OSCE was high, and feedback improved subsequent performance. CONCLUSIONS: Although internal and family medicine residents require additional training in specialized substance abuse skills, immediate feedback provided during an OSCE helped teach needed skills for assessing and managing substance abuse disorders. PMID- 16704388 TI - Competency in chest radiography. A comparison of medical students, residents, and fellows. AB - BACKGROUND: Accurate interpretation of chest radiographs (CXR) is essential as clinical decisions depend on readings. OBJECTIVE: We sought to evaluate CXR interpretation ability at different levels of training and to determine factors associated with successful interpretation. DESIGN: Ten CXR were selected from the teaching file of the internal medicine (IM) department. Participants were asked to record the most important diagnosis, their certainty in that diagnosis, interest in a pulmonary career and adequacy of CXR training. Two investigators independently scored each CXR on a scale of 0 to 2. PARTICIPANTS: Participants (n=145) from a single teaching hospital were third year medical students (MS) (n=25), IM interns (n=44), IM residents (n=45), fellows from the divisions of cardiology and pulmonary/critical care (n=16), and radiology residents (n=15). RESULTS: The median overall score was 11 of 20. An increased level of training was associated with overall score (MS 8, intern 10, IM resident 13, fellow 15, radiology resident 18, P<.001). Overall certainty was significantly correlated with overall score (r=.613, P<.001). Internal medicine interns and residents interested in a pulmonary career scored 14 of 20 while those not interested scored 11 (P=.027). Pneumothorax, misplaced central line, and pneumoperitoneum were diagnosed correctly 9%, 26%, and 46% of the time, respectively. Only 20 of 131 (15%) participants felt their CXR training sufficient. CONCLUSION: We identified factors associated with successful CXR interpretation, including level of training, field of training, interest in a pulmonary career and overall certainty. Although interpretation improved with training, important diagnoses were missed. PMID- 16704389 TI - Perceived, actual, and desired knowledge regarding Medicare billing and reimbursement. A national needs assessment survey of internal medicine residents. AB - BACKGROUND: Economics and reimbursement have become a daily part of practicing physicians' lives. Yet, few internal medicine (IM) programs have offered formal curricula during residency about practice management or economics. OBJECTIVE: To determine perceived, desired, and actual knowledge of Medicare billing and reimbursement among residents compared with community-based General Internists. DESIGN AND PARTICIPANTS: Cross-sectional needs assessment survey of community and university-based second-year IM residents from 4 geographic regions of the United States. RESULTS: One hundred and thirty-three second-year IM residents completed the questionnaire. Residents rated their level of knowledge about Medicare as a 2.0 (SD=0.9) on a Likert scale (1="very low," 5="very high"). Residents agreed that Medicare reimbursement should be taught in residency with a score of 4.0 (SD=1.1; 1="strongly disagree," 5="strongly agree" SD=1.1). On the knowledge assessment portion of the questionnaire, residents scored significantly lower than a group of general IM physicians who completed the same questions (percent correct=41.8% vs 59.0%, P<.001). Residents' scores correlated with their self assessed level of knowledge (P=.007). CONCLUSIONS: Our study demonstrates that second year IM residents feel they have a low level of knowledge regarding outpatient Medicare billing, and have a lower test score than practicing Internists to back up their feelings. The residents also strongly agree that they do not receive enough education about Medicare reimbursement, and believe it should be a requirement in residency training. PMID- 16704390 TI - Effect of the inpatient general medicine rotation on student pursuit of a generalist career. AB - BACKGROUND: Entry into general internal medicine (GIM) has declined. The effect of the inpatient general medicine rotation on medical student career choices is uncertain. OBJECTIVE: To assess the effect of student satisfaction with the inpatient general medicine rotation on pursuit of a career in GIM. DESIGN: Multicenter cohort study. PARTICIPANTS: Third-year medical students between July 2001 and June 2003. MEASUREMENTS: End-of-internal medicine clerkship survey assessed satisfaction with the rotation using a 5-point Likert scale. Pursuit of a career in GIM defined as: (1) response of "Very Likely" or "Certain" to the question "How likely are you to pursue a career in GIM?"; and (2) entry into an internal medicine residency using institutional match data. RESULTS: Four hundred and two of 751 (54%) students responded. Of the student respondents, 307 (75%) matched in the 2 years following their rotations. Twenty-eight percent (87) of those that matched chose an internal medicine residency. Of these, 8% (25/307) were pursuing a career in GIM. Adjusting for site and preclerkship interest, overall satisfaction with the rotation predicted pursuit of a career in GIM (odds ratio [OR] 3.91, P<.001). Although satisfaction with individual items did not predict pursuit of a generalist career, factor analysis revealed 3 components of satisfaction (attending, resident, and teaching). Adjusting for preclerkship interest, 2 factors (attending and teaching) were associated with student pursuit of a career in GIM (P<.01). CONCLUSIONS: Increased satisfaction with the inpatient general medicine rotation promotes pursuit of a career in GIM. PMID- 16704391 TI - Internal medicine residents' perceptions of cross-cultural training. Barriers, needs, and educational recommendations. AB - BACKGROUND: Physicians increasingly face the challenge of managing clinical encounters with patients from a range of cultural backgrounds. Despite widespread interest in cross-cultural care, little is known about resident physicians' perceptions of what will best enable them to provide quality care to diverse patient populations. OBJECTIVES: To assess medicine residents' (1) perceptions of cross-cultural care, (2) barriers to care, and (3) training experiences and recommendations. DESIGN, SETTING, AND PATIENTS: Qualitative individual interviews were conducted with 26 third-year medicine residents at Massachusetts General Hospital in Boston (response rate=87%). Interviews were recorded, transcribed, and analyzed. RESULTS: Despite significant interest in cross-cultural care, almost all of the residents reported very little training during residency. Most had gained cross-cultural skills through informal learning. A few were skeptical about formal training, and some expressed concern that it is impossible to understand every culture. Challenges to the delivery of cross-cultural care included managing patients with limited English proficiency, who involve family in critical decision making, and who have beliefs about disease that vary from the biomedical model. Residents cited many implications to these barriers, ranging from negatively impacting the patient-physician relationship to compromised care. Training recommendations included making changes to the educational climate and informal and formal training mechanisms. CONCLUSIONS: If cross-cultural education is to be successful, it must take into account residents' perspectives and be focused on overcoming residents' cited barriers. It is important to convey that cross-cultural education is a set of skills that can be taught and applied, in a time-efficient manner, rather than requiring an insurmountable knowledge base. PMID- 16704392 TI - Spirituality and medicine. A workshop for medical students and residents. AB - INTRODUCTION: Governing bodies for medical education recommend that spirituality and medicine be incorporated into training. AIM: To pilot a workshop on spirituality and medicine on a convenience sample of preclinical medical students and internal medicine residents and determine whether content was relevant to learners at different levels, whether preliminary evaluation was promising, and to generate hypotheses for future research. SETTING: Private medical school and university primary care internal medicine residency program, both in the Northeast. CURRICULUM DESCRIPTION: The authors designed and implemented a required 2-hour workshop for all second-year medical students and a separate required 1.5-hour workshop for all primary care internal medicine house staff. The workshops used multiple educational strategies including lecture, discussion, and role-play to address educational objectives. PROGRAM EVALUATION: Learners completed optional, anonymous pre and postworkshop surveys with six 5-point Likert-rated statements and space to cite the most useful part of the curriculum and their remaining questions. One hundred and thirty-seven learners participated and 100 completed both surveys. Medical students and residents had increased (all P< or =.002): agreement regarding the appropriateness of inquiring about spiritual and religious beliefs in the medical encounter, their perceived competence in taking a spiritual history, and their perceived knowledge of available pastoral care resources. Medical students, but not residents, had an increase in their perceived comfort in working with hospital chaplains. DISCUSSION: A brief pilot workshop on spirituality and medicine had a modest effect in improving attitudes and perceived competence of both medical students and residents. PMID- 16704394 TI - Brief report: Brief instrument to assess geriatrics knowledge of surgical and medical subspecialty house officers. AB - PROBLEM: Initiatives are underway to increase geriatrics training in nonprimary care disciplines. However, no validated instrument exists to measure geriatrics knowledge of house officers in surgical specialties and medical subspecialties. METHODS: A 23-item multiple-choice test emphasizing inpatient care and common geriatric syndromes was developed through expert panels and pilot testing, and administered to 305 residents and fellows at 4 institutions in surgical disciplines (25% of respondents), emergency medicine (29%), medicine subspecialties (19%), internal medicine (12%), and other disciplines (15%). RESULTS: Three items decreased internal reliability. The remaining 20 items covered 17 topic areas. Residents averaged 62% correct on the test. Internal consistency was appropriate (Cronbach's alpha coefficient=0.60). Validity was supported by the use of expert panels to develop content, and by overall differences in scores by level of training (P<.0001) and graded improvement in test performance, with 58%, 63%, 62%, and 69% correct responses among HO1, HO2, HO3, and HO4s, respectively. CONCLUSIONS: This reliable, valid measure of clinical geriatrics knowledge can be used by a wide variety of surgical and medical graduate medical education programs to guide curriculum reform or evaluate program performance to meet certification requirements. The instrument is now available on the web. PMID- 16704393 TI - Brief report: Multiprogram evaluation of reading habits of primary care internal medicine residents on ambulatory rotations. AB - OBJECTIVE: To assess the reading habits and educational resources of primary care internal medicine residents for their ambulatory medicine education. DESIGN: Cross-sectional, multiprogram survey of primary care internal medicine residents. PARTICIPANTS/SETTING: Second- and third-year residents on ambulatory care rotations at 9 primary care medicine programs (124 eligible residents; 71% response rate). MEASUREMENTS AND MAIN RESULTS: Participants were asked open-ended and 5-point Likert-scaled questions about reading habits: time spent reading, preferred resources, and motivating and inhibiting factors. Participants reported reading medical topics for a mean of 4.3+/-3.0 SD hours weekly. Online-only sources were the most frequently utilized medical resource (mean Likert response 4.16+/-0.87). Respondents most commonly cited specific patients' cases (4.38+/ 0.65) and preparation for talks (4.08+/-0.89) as motivating factors, and family responsibilities (3.99+/-0.65) and lack of motivation (3.93+/-0.81) as inhibiting factors. CONCLUSIONS: To stimulate residents' reading, residency programs should encourage patient- and case-based learning; require teaching assignments; and provide easy access to online curricula. PMID- 16704395 TI - Brief report: Internal medicine residents', attendings', and nurses' perceptions of the night float system. AB - BACKGROUND: Residency training programs use the night float system increasingly to meet the new resident work hour regulations. OBJECTIVE: To assess and compare residents', attendings', and nurses' perceptions of the night float system. DESIGN: A survey study. PARTICIPANTS: One hundred and seven residents, 48 attendings, and 69 nurses in a university-based multicenter internal medicine residency program. MEASUREMENTS: Perceived impact on patient care, resident training, and resident performance. RESULTS: The overall response rate was 75%. In general, more residents than both attendings and nurses had positive opinions regarding the night float system, particularly in relation to patient care. Only a small proportion of residents and attendings thought positively about the night float's impact on training quality (29.9%; 18.2%), daily feedback (23.0%; 9.1%), and end of rotation evaluation (21.8%; 6.1%). Less than half of the nurses had positive perceptions of the night residents' performance in terms of promptness (40.9%), physical availability (38.6%), familiarity with the patients' cases, and management plans (15.9%), communication of management plans to nurses (36.4%), professional respect and trust (43.2%), and teamwork (45.5%). CONCLUSIONS: Residents had more positive perceptions than attendings and nurses. Nurses, in particular, had negative perceptions of resident performance in the setting of the night float system. PMID- 16704397 TI - Brief report: Use of the mini-clinical evaluation exercise in internal medicine core clerkships. AB - INTRODUCTION: Direct observation of medical students' clinical skills is important, but occurs infrequently. The mini-clinical evaluation exercise (mCEX) is a tool developed for use with internal medicine (IM) residents that can be used to promote direct observation of medical students' clinical skills. It is unknown how many IM core clerkships in the United States use the mCEX or how it has been implemented. METHODS: Questions about use of the mCEX were incorporated into an online annual survey distributed to the 114 IM clerkships belonging to Clerkship Directors in Internal Medicine, a national organization of individuals responsible for teaching IM to medical students. RESULTS: The survey response rate was 83%. Twenty-eight percent (N=27) of respondents use the mCEX in their clerkship. The mean number of required mCEX encounters is 2.3 (SD 1.6). The mCEX is used for formative assessment (68%) more than summative assessment (11%). Ward attendings are the most common mCEX evaluators (72%). DISCUSSION: The mCEX is being used to promote direct observation of medical students' clinical skills in a significant minority of IM core clerkships. The mCEX is 1 tool for facilitating feedback from both faculty and residents on trainees' developing skills. PMID- 16704396 TI - Brief report: Reducing inappropriate usage of stress ulcer prophylaxis among internal medicine residents. A practice-based educational intervention. AB - BACKGROUND: Many inpatients receive stress ulcer prophylaxis (SUP) inappropriately. This indiscriminate usage increases costs and avoidable side effects. Practice-based learning and improvement (PBLI) methodology may improve compliance with published guidelines. OBJECTIVE: To investigate the response of internal medicine residents to an educational intervention regarding SUP. DESIGN: A prospective, pre and postintervention cohort study using an educational intervention based on PBLI. PATIENTS: Three groups of consecutively admitted patients (1 group preintervention and 2 groups postintervention) on the medicine ward at a University Hospital. MAIN OUTCOME MEASURE: Rates of inappropriate SUP prescription and discharge with an inappropriate prescription. RESULTS: One month after the intervention, inappropriate prophylaxis was significantly decreased (59% pre, 29% postintervention, P<.002). The rate of discharge with an inappropriate prescription also decreased, but was not significant (25% pre, 14% postintervention, P=.14). In the 6-month postintervention cohort, inappropriate SUP remained lower (59% pre, 33% postintervention, P<.007). The rate of discharge with an inappropriate prescription was also significantly lower (25% pre, 7% postintervention, P<.009). CONCLUSION: Practice-based learning and improvement can improve compliance with published guidelines, and change practice patterns. After the intervention, both inappropriate prophylaxis and inappropriate prescriptions upon discharge were reduced. Importantly, the intervention was sustained, transmitted across academic years to a new class of interns who had not directly experienced the intervention. PMID- 16704398 TI - Brief report: Utilizing an audiotape for outpatient preceptor faculty development. AB - BACKGROUND: Faculty development for busy and geographically dispersed ambulatory preceptors is a difficult task for course directors. PURPOSE: A faculty development audiotape intended for playing in the preceptor's car was created. The feasibility of this form of faculty development was tested in this pilot study. METHODS: A short audiotape, focusing on strategies for the provision of independence to students in the office setting, was made and distributed to all preceptors of students taking a fourth-year required clerkship in ambulatory medicine. Preceptor behavior was reported by students on postclerkship evaluations before and after tape distribution. RESULTS: In the year before tape distribution, 21% of evaluations indicated a lack of independence on the part of the student, compared with 14% in the year following the intervention (P=.03). There was no regression of behavior among preceptors already providing independence. Among the preceptors initially identified as not following recommendations for student independence, the percentage of evaluations indicating a lack of independence went from 72% preintervention to 42% postintervention (P<.001). CONCLUSIONS: A short audiotape is a novel form of faculty development, which was acceptable to preceptors and may influence teaching behavior in the desired manner. PMID- 16704399 TI - The PRIME curriculum. Clinical research training during residency. AB - AIM: The Primary Medical Education (PRIME) program is an outpatient-based, internal medicine residency track nested within the University of California, San Francisco (UCSF) categorical medicine program. Primary Medical Education is based at the San Francisco Veteran's Affairs Medical Center (VAMC), 1 of 3 teaching hospitals at UCSF. The program accepts 8 UCSF medicine residents annually, who differentiate into PRIME after internship. In 2000, we implemented a novel research methods curriculum with the dual purposes of teaching basic epidemiology skills and providing mentored opportunities for clinical research projects during residency. SETTING: Single academic internal medicine program. PROGRAM DESCRIPTION: The PRIME curriculum utilizes didactic lecture, frequent journal clubs, work-in-progress sessions, and active mentoring to enable residents to "try out" a clinical research project during residency. PROGRAM EVALUATION: Among 32 residents in 4 years, 22 residents have produced 20 papers in peer-reviewed journals, 1 paper under review, and 2 book chapters. Their clinical evaluations are equivalent to other UCSF medicine residents. DISCUSSION: While learning skills in evidence-based medicine, residents can conduct high-quality research. Utilizing a collaboration of General Internal Medicine researchers and educators, our curriculum affords residents the opportunity to "try-out" clinical research as a potential future career choice. PMID- 16704400 TI - "Oh! She doesn't speak English!" Assessing resident competence in managing linguistic and cultural barriers. AB - BACKGROUND: Residents must master complex skills to care for culturally and linguistically diverse patients. METHODS: As part of an annual 10-station, standardized patient (SP) examination, medical residents interacted with a 50 year-old reserved, Bengali-speaking woman (SP) with a positive fecal occult blood accompanied by her bilingual brother (standardized interpreter (SI)). While the resident addressed the need for a colonoscopy, the SI did not translate word for word unless directed to, questioned medical terms, and was reluctant to tell the SP frightening information. The SP/SI, faculty observers, and the resident assessed the performance. RESULTS: Seventy-six residents participated. Mean faculty ratings (9-point scale) were as follows: overall 6.0, communication 6.0, knowledge 6.3. Mean SP/SI ratings (3.1, range 1.9 to 3.9) correlated with faculty ratings (overall r=.719, communication r=.639, knowledge r=.457, all P<.01). Internal reliability as measured by Cronbach's alpha coefficients for the 20 item instrument was 0.91. Poor performance on this station was associated with poor performance on other stations. Eighty-nine percent of residents stated that the educational value was moderate to high. CONCLUSION: We reliably assessed residents communication skills conducting a common clinical task across a significant language barrier. This medical education innovation provides the first steps to measuring interpreter facilitated skills in residency training. PMID- 16704401 TI - The use of tissue models for vascular access training. Phase I of the procedural patient safety initiative. AB - INTRODUCTION: Following the Institute of Medicine report "To Err is Human," the Agency for Healthcare Research and Quality identified proper central venous catheter (CVC) insertion techniques and wide sterile barriers (WSB) as 2 major quality indicators for patient safety. However, no standard currently exists to teach proper procedural techniques to physicians. AIM: To determine whether our nonhuman tissue model is an effective tool for teaching physicians proper wide sterile barrier technique, ultrasound guidance for CVC placement, and sharps safety. PARTICIPANTS: Educational sessions were organized for physicians at Cedars-Sinai Medical Center. Participants had a hands-on opportunity to practice procedural skills using a nonhuman tissue model, under the direct supervision of experienced proceduralists. PROGRAM EVALUATION: An anonymous survey was distributed to participants both before and after training, measuring their reactions to all aspects of the educational sessions relative to their prior experience level. DISCUSSION: The sessions were rated highly worthwhile, and statistically significant improvements were seen in comfort levels with ultrasound-guided vascular access and WSB (P<.001). Given the revitalized importance of patient safety and the emphasis on reducing medical errors, further studies on the utility of nonhuman tissue models for procedural training should be enthusiastically pursued. PMID- 16704402 TI - Preparing fourth-year medical students to teach during internship. AB - Interns are expected to teach medical students, yet there is little formal training in medical school to prepare them for this role. To enhance the teaching skills of our graduating students we initiated a 4-hour "teaching to teach" course as part of the end of the fourth-year curriculum. Course evaluations demonstrate that students strongly support this program (overall ratings 2000 to 2005: mean=4.4 [scale 1 to 5], n=224). When 2004 course participants were surveyed during the last month of their internship, 84%"agree" or "strongly agree" with the statement: "The teaching to teach course helped prepare me for my role as a teacher during internship" (2005: mean 4.2 [scale 1 to 5], n=45, response rate 60%). A course preparing fourth-year students to teach during internship is both feasible and reproducible, with a minimal commitment of faculty and resident time. Participants identify it as an important addition to their education and as useful during internship. PMID- 16704403 TI - Musculoskeletal exam and joint injection training for internal medicine residents. AB - We developed a musculoskeletal education intervention for internal medicine residents consisting of lectures, anatomic models, and a joint injection clinic. Written tests, observed musculoskeletal exams, and self-confidence scales were administered to 27 internal medicine residents before and after the intervention. Improvement was found in observed physical exam and self-reported confidence levels in performing knee injections. Confidence in shoulder injection skills improved, but remained low. Improved scores on written examination, though statistically significant, were not educationally significant. This single institution pilot study demonstrates that a simple educational program results in improved knee and shoulder examination skills and confidence in performing knee injections. PMID- 16704404 TI - Educating for empathy. A review. AB - OBJECTIVE: Empathy in the medical setting is appreciation of the patient's emotions and expression of that awareness to the patient. Named as an essential learning objective by the American Association of Medical Colleges, empathy is believed to significantly influence patient satisfaction, adherence to medical recommendations, clinical outcomes, and professional satisfaction. The objective of this study was to identify effective strategies to enhance empathy in undergraduate medical students. DATA SOURCES: We searched PubMed for studies that address the effectiveness of strategies for teaching empathy to medical students. We identified 13 peer-reviewed, English language, qualitative and quantitative studies reporting primary data on interventions that aim to foster empathy in undergraduate medical students, using Medical Subject Heading terms education, medical, undergraduate or student, medical crossed with empathy. RESULTS: These studies indicate that empathy may be amenable to positive change with a range of interventional strategies. Communication skill workshops addressing the behavioral dimension of empathy show greatest quantitative impact on participants. However, current studies are challenged by varying definitions of empathy, small sample sizes, lack of adequate control groups, and variation among existing empathy measurement instruments. CONCLUSION: Given the methodological limitations of the available studies, and uncertainty about which dimensions of empathy should be addressed, larger studies using validated measurement tools are recommended. PMID- 16704405 TI - Use of handheld computers in medical education. A systematic review. AB - BACKGROUND: Over the past decade, handheld computers (or personal digital assistants [PDAs]) have become a popular tool among medical trainees and physicians. Few comprehensive reviews of PDA use in medicine have been published. OBJECTIVE: We systematically reviewed the literature to (1) describe medical trainees' use of PDAs for education or patient care, (2) catalog popular software applications, and (3) evaluate the impact of PDA use on patient care. DATA SOURCES: MEDLINE (1993 to 2004), medical education-related conference proceedings, and hand search of article bibliographies. REVIEW METHODS: We identified articles and abstracts that described the use of PDAs in medical education by trainees or educators. Reports presenting a qualitative or quantitative evaluation were included. RESULTS: Sixty-seven studies met inclusion criteria. Approximately 60% to 70% of medical students and residents use PDAs for educational purposes or patient care. Satisfaction was generally high and correlated with the level of handheld computer experience. Most of the studies included described PDA use for patient tracking and documentation. By contrast, trainees rated medical textbooks, medication references, and medical calculators as the most useful applications. Only 1 randomized trial with educational outcomes was found, demonstrating improved learning and application of evidence based medicine with use of PDA-based decision support software. No articles reported the impact of PDA use on patient outcomes. CONCLUSION: Most medical trainees find handhelds useful in their medical education and patient care. Further studies are needed to evaluate how PDAs impact learning and clinical outcomes. PMID- 16704406 TI - Beyond journal clubs. Moving toward an integrated evidence-based medicine curriculum. AB - Incorporating evidence-based medicine (EBM) into clinical practice is an important competency that residency training must address. Residency program directors, and the clinical educators who work with them, should develop curricula to enhance residents' capacity for independent evidence-based practice. In this article, the authors argue that residency programs must move beyond journal club formats to promote the practice of EBM by trainees. The authors highlight the limitations of journal club, and suggest additional curricular approaches for an integrated EBM curriculum. Helping residents become effective evidence users will require a sustained effort on the part of residents, faculty, and their educational institutions. PMID- 16704407 TI - Are we making progress in medical education? PMID- 16704408 TI - The persistent myth of stability. On the chronic underestimation of the role of context in behavior. PMID- 16704409 TI - Nitric oxide and cell proliferation. AB - Nitric oxide (NO*) has been proposed to be a physiological modulator of cell proliferation, able to promote in most cases cell cycle arrest. In this review I explore the molecular basis of this mechanism of action. The modulatory action of NO* on the intracellular concentration of cGMP and the machinery directly involved in the control of cell cycle progression, including the expression and activity of diverse cyclins and cyclin-dependent kinases, their physiological inhibitors, and the master transcriptional regulator retinoblastoma protein, will be discussed. The role of NO* in proliferation mediated by tyrosine kinase receptors such as the epidermal growth factor receptor and downstream signalling pathways will also be considered. Finally, the involvement of NO* in proliferative processes relevant for normal development will be outlined. PMID- 16704410 TI - Insulin induces heme oxygenase-1 through the phosphatidylinositol 3-kinase/Akt pathway and the Nrf2 transcription factor in renal cells. AB - Heme oxygenase-1 catalyzes the breakdown of heme and is protective in models of kidney transplantation. In this study we describe the induction of heme oxygenase 1 mRNA and protein by insulin. Following treatment with insulin, a five-fold increase in heme oxygenase-1 mRNA and a four-fold increase in protein expression were observed in renal adenocarcinoma cells; insulin-induced heme oxygenase-1 expression was also demonstrated in mouse primary tubular epithelial cells. The induction of heme oxygenase-1 in renal adenocarcinoma cells was blocked by actinomycin D and cycloheximide and was abolished by the phosphatidylinositol 3 kinase inhibitor, LY294002, but not by the inactive analog LY303511. Overexpressing a dominant-negative form of Akt abrogated the heme oxygenase-1 inducing effects of insulin, whereas cells transfected with a constitutively active Akt construct demonstrated an increase in heme oxygenase-1 promoter activity and protein expression. The transcription factor NF-E2-related factor-2 was found to translocate to the nucleus following insulin treatment in a phosphatidylinositol 3-kinase-dependent manner. Pretreatment with NF-E2-related factor-2 small-interfering RNA abolished insulin-induced heme oxygenase-1 induction. Insulin was also found to activate the mitogen-activated protein kinase cascades p38 and extracellular signal-related kinase; however, inhibition of these pathways with SB202190 and PD98059 did not alter insulin-induced heme oxygenase-1 expression. Thus, insulin induces heme oxygenase-1 mRNA and protein expression in renal cells in a phosphatidylinositol 3-kinase/Akt and NF-E2 related factor-2-dependent manner. PMID- 16704411 TI - The beta domain is required for Vps4p oligomerization into a functionally active ATPase. AB - Endocytic and biosynthetic trafficking pathways to the lysosome/vacuole converge at the prevacuolar endosomal compartment. During transport through this compartment, integral membrane proteins that are destined for delivery to the lysosome/vacuole lumen undergo multivesicular body (MVB) sorting into internal vesicles formed by invagination of the endosomal limiting membrane. Vps4 is an AAA family ATPase which plays a key role in MVB sorting and facilitates transport through endosomes. It possesses an N-terminal microtubule interacting and trafficking domain required for recruitment to endosomes and an AAA domain with an ATPase catalytic site. The recently solved 3D structure revealed a beta domain, which protrudes from the AAA domain, and a final C-terminal alpha-helix. However, the in vivo roles of these domains are not known. In this study, we have identified motifs in these domains that are highly conserved between yeast and human Vps4. We have mutated these motifs and studied the effect on yeast Vps4p function in vivo and in vitro. We show that the beta domain of the budding yeast Vps4p is not required for recruitment to endosomes, but is essential for all Vps4p endocytic functions in vivo. We also show that the beta domain is required for Vps4p homotypic interaction and for full ATPase activity. In addition, it is required for interaction with Vta1p, which works in concert with Vps4p in vivo. Our studies suggest that assembly of a Vps4p oligomeric complex with full ATPase activity that interacts with Vta1p is essential for normal endosome function. PMID- 16704412 TI - Characterization of a novel long-chain acyl-CoA thioesterase from Alcaligenes faecalis. AB - A novel long-chain acyl-CoA thioesterase from Alcaligenes faecalis has been isolated and characterized. The protein was extracted from the cells with 1 m NaCl, which required 1.5-fold, single-step purification to yield near-homogeneous preparations. In solution, the protein exists as homomeric aggregates, of mean diameter 21.6 nm, consisting of 22-kDa subunits. MS/MS data for peptides obtained by trypsin digestion of the thiosterase did not match any peptide from Escherichia coli thioesterases or any other thioesterases in the database. The thioesterase was associated exclusively with the surface of cells as revealed by ultrastructural studies using electron microscopy and immunogold labeling. It hydrolyzed saturated and unsaturated fatty acyl-CoAs of C12 to C18 chain length with Vmax and Km of 3.58-9.73 micromol x min(-1) x (mg protein)(-1) and 2.66-4.11 microm, respectively. A catalytically important histidine residue is implicated in the active site of the enzyme. The thioesterase was active and stable over a wide range of temperature and pH. Maximum activity was observed at 65 degrees C and pH 10.5, and varied between 60% and 80% at temperatures of 25-70 degrees C and pH 6.5-10. The thioesterase also hydrolyzed p-nitrophenyl esters of C2 to C12 chain length, but substrate competition experiments demonstrated that the long chain acyl-CoAs are better substrates for thioesterase than p-nitrophenyl esters. When assayed at 37 and 20 degrees C, the affinity and catalytic efficiency of the thioesterase for palmitoleoyl-CoA and cis-vaccenoyl-CoA were reduced approximately twofold at the lower temperature, but remained largely unaltered for palmitoyl-CoA. PMID- 16704413 TI - Constitutively active alpha subunits of G(q/11) and G(12/13) families inhibit activation of the pro-survival Akt signaling cascade. AB - Accumulating evidence indicates that G protein signaling plays an active role in the regulation of cell survival. Our previous study demonstrated the regulatory effects of G(i/o) proteins in nerve growth factor-induced activation of pro survival Akt kinase. In the present study we explored the role of various members of the G(s), G(q/11) and G(12/13) subfamilies in the regulation of Akt in cultured mammalian cells. In human embryonic kidney 293 cells transiently expressing constitutively active mutants of G alpha11, G alpha14, G alpha16, G alpha12, or G alpha13 (G alpha11QL, G alpha14QL, G alpha16QL, G alpha12QL and G alpha13QL, respectively), basal phosphorylation of Akt was attenuated, as revealed by western blotting analysis using a phosphospecific anti-Akt immunoglobulin. In contrast, basal Akt phosphorylation was unaffected by the overexpression of a constitutively active G alpha(s) mutant (G alpha(s)QL). Additional experiments showed that G alpha11QL, G alpha14QL, G alpha16QL, G alpha12QL and G alpha13QL, but not G alpha(s)QL, attenuated phosphorylation of the Akt-regulated translation regulator tuberin. Moreover, they were able to inhibit the epidermal growth factor-induced Akt activation and tuberin phosphorylation. The inhibitory mechanism of Gq family members was independent of phospholipase Cbeta activation and calcium signaling because G alpha11QL, G alpha14QL and G alpha16QL remained capable of inhibiting epidermal growth factor induced Akt activation in cells pretreated with U73122 and the intracellular calcium chelator, BAPTA/AM. Finally, overexpression of the dominant negative mutant of RhoA blocked G alpha12QL- and G alpha13QL-mediated inhibition, suggesting that activated G alpha12 and G alpha13 inhibit Akt signaling via RhoA. Collectively, this study demonstrated the inhibitory effect of activated G alpha11, G alpha14, G alpha16, G alpha12 and G alpha13 on pro-survival Akt signaling. PMID- 16704414 TI - Zinc potentiates the antibacterial effects of histidine-rich peptides against Enterococcus faecalis. AB - Antimicrobial peptides are effector molecules of the innate immune system. We have recently shown that peptides containing multiples of the heparin-binding Cardin and Weintraub motifs AKKARA and ARKKAAKA exert antimicrobial activities. Here, we show that replacement of lysine and arginine in these motifs by histidine abrogates the antibacterial effects of these peptides. Antibacterial activity of the histidine-rich peptides against the Gram-positive bacterium Enterococcus faecalis was restored by the addition of Zn2+. Fluorescence microscopy experiments showed that Zn2+ enabled binding of the histidine-rich peptides to Enterococcus faecalis bacteria. Similar Zn2+-dependent antibacterial activities were shown for histatin 5 as well as histidine-containing peptides derived from the Zn2+- and heparin-binding domain 5 of human kininogen. Thus, the results demonstrate a previously undisclosed Zn2+-dependent antibacterial activity of kininogen-derived peptides and indicate an important role for Zn2+ in regulating the antimicrobial activities of histidine-rich peptides. PMID- 16704415 TI - Structural basis for the recognition of complex-type biantennary oligosaccharides by Pterocarpus angolensis lectin. AB - The crystal structure of Pterocarpus angolensis lectin is determined in its ligand-free state, in complex with the fucosylated biantennary complex type decasaccharide NA2F, and in complex with a series of smaller oligosaccharide constituents of NA2F. These results together with thermodynamic binding data indicate that the complete oligosaccharide binding site of the lectin consists of five subsites allowing the specific recognition of the pentasaccharide GlcNAc beta(1-2)Man alpha(1-3)[GlcNAc beta(1-2)Man alpha(1-6)]Man. The mannose on the 1 6 arm occupies the monosaccharide binding site while the GlcNAc residue on this arm occupies a subsite that is almost identical to that of concanavalin A (con A). The core mannose and the GlcNAc beta(1-2)Man moiety on the 1-3 arm on the other hand occupy a series of subsites distinct from those of con A. PMID- 16704416 TI - Novel beta-1,3-, 1,6-oligoglucan elicitor from Alternaria alternata 102 for defense responses in tobacco. AB - A novel elicitor that induces chitinases in tobacco BY-2 cells was isolated from Alternaria alternata 102. Six other fungi, including A. alternata IFO 6587, could not induce, or weakly induce chitinase activity. The purified elicitor was soluble in 75% methanol and showed the chitinase-inducing activity when applied at concentrations of as low as 25 ng x mL(-1). Structural determination by methylation analysis, reducing-end analysis, MALDI-TOF/MS, and NMR spectroscopy indicated that the elicitor was a mixture of beta-1,3-, 1,6-oligoglucans mostly with a degree of polymerization of between 8 and 17. Periodate oxidation of the elicitor suggested that the 1,6-linked and nonreducing terminal residues are essential for the elicitor activity. Further analysis of the elicitor responses in BY-2 cells indicated that the activity of this beta-1,3-, 1,6-glucan elicitor was about 1000 times more potent than that of laminarin, which is a known elicitor of defense responses in tobacco. Analyzing the expression of defense related genes indicated that a phenylalanine ammonia-lyase gene and a coumaroyl CoA O-methyltransferase gene were transiently expressed by this beta-1,3-, 1,6 glucan elicitor. The elicitor induced a weak oxidative burst but did not induce cell death in the BY-2 cells. In the tissue of tobacco plants, this beta-1,3-, 1,6-glucan elicitor induced the expression of basic PR-3 genes, the phenylpropanoid pathway genes, and the sesquiterpenoid pathway genes. In comparison with laminarin and laminarin sulfate, which are reported to be potent elicitors of defense responses in tobacco, the expression pattern of genes induced by the purified beta-1,3-, 1,6-glucan elicitor was more similar to that induced by laminarin than to that induced by laminarin sulfate. PMID- 16704417 TI - Comparative biochemical characterization of nitrile-forming proteins from plants and insects that alter myrosinase-catalysed hydrolysis of glucosinolates. AB - The defensive function of the glucosinolate-myrosinase system in plants of the order Capparales results from the formation of isothiocyanates when glucosinolates are hydrolysed by myrosinases upon tissue damage. In some glucosinolate-containing plant species, as well as in the insect herbivore Pieris rapae, protein factors alter the outcome of myrosinase-catalysed glucosinolate hydrolysis, leading to the formation of products other than isothiocyanates. To date, two such proteins have been identified at the molecular level, the epithiospecifier protein (ESP) from Arabidopsis thaliana and the nitrile specifier protein (NSP) from P. rapae. These proteins share no sequence similarity although they both promote the formation of nitriles. To understand the biochemical bases of nitrile formation, we compared some of the properties of these proteins using purified preparations. We show that both proteins appear to be true enzymes rather than allosteric cofactors of myrosinases, based on their substrate and product specificities and the fact that the proportion of glucosinolates hydrolysed to nitriles does not remain constant when myrosinase activity varies. No stable association between ESP and myrosinase could be demonstrated during affinity chromatography, nevertheless some proximity of ESP to myrosinase is required for epithionitrile formation to occur, as evidenced by the lack of ESP activity when it was spatially separated from myrosinase in a dialysis chamber. The significant difference in substrate- and product specificities between A. thaliana ESP and P. rapae NSP is consonant with their different ecological functions. Furthermore, ESP and NSP differ remarkably in their requirements for metal ion cofactors. We found no indications of the involvement of a free radical mechanism in epithionitrile formation by ESP as suggested in earlier reports. PMID- 16704418 TI - Extraenzymatic functions of the dipeptidyl peptidase IV-related proteins DP8 and DP9 in cell adhesion, migration and apoptosis. AB - The dipeptidyl peptidase IV gene family contains the four peptidases dipeptidyl peptidase IV, fibroblast activation protein, dipeptidyl peptidase 8 and dipeptidyl peptidase 9. Dipeptidyl peptidase IV and fibroblast activation protein are involved in cell-extracellular matrix interactions and tissue remodeling. Fibroblast activation protein is upregulated and dipeptidyl peptidase IV is dysregulated in chronic liver disease. The effects of dipeptidyl peptidase 8 and dipeptidyl peptidase 9 on cell adhesion, cell migration, wound healing and apoptosis were measured by using green fluorescent protein fusion proteins to identify transfected cells. Dipeptidyl peptidase 9-overexpressing cells exhibited impaired cell adhesion, migration in transwells and monolayer wound healing on collagen I, fibronectin and Matrigel. Dipeptidyl peptidase 8-overexpressing cells exhibited impaired cell migration on collagen I and impaired wound healing on collagen I and fibronectin in comparison to the green fluorescent protein transfected controls. Dipeptidyl peptidase 8 and dipeptidyl peptidase 9 enhanced induced apoptosis, and dipeptidyl peptidase 9 overexpression increased spontaneous apoptosis. Mechanistic investigations showed that neither the catalytic serine of dipeptidyl peptidase 8 or dipeptidyl peptidase 9 nor the Arg Gly-Asp integrin-binding motif in dipeptidyl peptidase 9 were required for the impairment of cell survival, cell adhesion or wound healing. We have previously shown that the in vitro roles of dipeptidyl peptidase IV and fibroblast activation protein in cell-extracellular matrix interactions and apoptosis are similarly independent of catalytic activity. Dipeptidyl peptidase 9 overexpression reduced beta-catenin, tissue inhibitor of matrix metalloproteinases 2 and discoidin domain receptor 1 expression. This is the first demonstration that dipeptidyl peptidase 8 and dipeptidyl peptidase 9 influence cell-extracellular matrix interactions, and thus may regulate tissue remodeling. PMID- 16704419 TI - Sugar and alcohol molecules provide a therapeutic strategy for the serpinopathies that cause dementia and cirrhosis. AB - Mutations in neuroserpin and alpha1-antitrypsin cause these proteins to form ordered polymers that are retained within the endoplasmic reticulum of neurones and hepatocytes, respectively. The resulting inclusions underlie the dementia familial encephalopathy with neuroserpin inclusion bodies (FENIB) and Z alpha1 antitrypsin-associated cirrhosis. Polymers form by a sequential linkage between the reactive centre loop of one molecule and beta-sheet A of another, and strategies that block polymer formation are likely to be successful in treating the associated disease. We show here that glycerol, the sugar alcohol erythritol, the disaccharide trehalose and its breakdown product glucose reduce the rate of polymerization of wild-type neuroserpin and the Ser49Pro mutant that causes dementia. They also attenuate the polymerization of the Z variant of alpha1 antitrypsin. The effect on polymerization was apparent even when these agents had been removed from the buffer. None of these agents had any detectable effect on the structure or inhibitory activity of neuroserpin or alpha1-antitrypsin. These data demonstrate that sugar and alcohol molecules can reduce the polymerization of serpin mutants that cause disease, possibly by binding to and stabilizing beta sheet A. PMID- 16704420 TI - Variants of beta-microglobulin cleaved at lysine-58 retain the main conformational features of the native protein but are more conformationally heterogeneous and unstable at physiological temperature. AB - Cleavage of the small amyloidogenic protein beta2-microglobulin after lysine-58 renders it more prone to unfolding and aggregation. This is important for dialysis-related beta2-microglobulin amyloidosis, since elevated levels of cleaved beta2-microglobulin may be found in the circulation of dialysis patients. However, the solution structures of these cleaved beta2-microglobulin variants have not yet been assessed using single-residue techniques. We here use such methods to examine beta2-microglobulin cleaved after lysine-58 and the further processed variant (found in vivo) from which lysine-58 is removed. We find that the solution stability of both variants, especially of beta2-microglobulin from which lysine-58 is removed, is much reduced compared to wild-type beta2 microglobulin and is strongly dependent on temperature and protein concentration. 1H-NMR spectroscopy and amide hydrogen (1H/2H) exchange monitored by MS show that the overall three-dimensional structure of the variants is similar to that of wild-type beta2-microglobulin at subphysiological temperatures. However, deviations do occur, especially in the arrangement of the B, D and E beta-strands close to the D-E loop cleavage site at lysine-58, and the experiments suggest conformational heterogeneity of the two variants. Two-dimensional NMR spectroscopy indicates that this heterogeneity involves an equilibrium between the native-like fold and at least one conformational intermediate resembling intermediates found in other structurally altered beta2-microglobulin molecules. This is the first single-residue resolution study of a specific beta2 microglobulin variant that has been found circulating in dialysis patients. The instability and conformational heterogeneity of this variant suggest its involvement in beta2-microglobulin amyloidogenicity in vivo. PMID- 16704421 TI - Electron-transfer capacity of catechin derivatives and influence on the cell cycle and apoptosis in HT29 cells. AB - Galloylated and nongalloylated catechin conjugates with cysteine derivatives have been synthesized and evaluated for their capacity to scavenge free radicals and to influence crucial functions (cell cycle, apoptosis) in HT29 colon carcinoma cells. We show that the nonphenolic part of the molecule modified the capacity of catechins to donate hydrogen atoms and to transfer electrons to free radicals. Nongalloylated derivatives did not significantly influence either the cell cycle or apoptosis. Among the galloylated species, 4beta-[S-(O-ethyl cysteinyl)]epicatechin 3-O-gallate, which showed a high electron-transfer capacity (5 e- per molecule), arrested the cell cycle and induced apoptosis as expected for galloylated catechins such as tea (-)-epigallocatechin 3-O-gallate. 4beta-[S-(N-Acetyl-O-methyl-cysteinyl)]epicatechin 3-O-gallate, which showed the highest hydrogen-donating capacity (10 H per molecule) while keeping the electron transfer capacity low (2.9 e- per molecule), did not trigger any significant apoptosis. The gallate moiety did not appear to be sufficient for the pro apoptotic effect of the catechin derivatives in HT29 cells. Instead, a high electron-transfer capacity is more likely to be behind this effect. The use of stable radicals sensitive exclusively to electron transfer may help to design molecules with either preventive scavenging action (high hydrogen donation, low electron transfer) or therapeutic pro-apoptotic activity (high electron transfer). PMID- 16704422 TI - The subcellular localization of vaccinia-related kinase-2 (VRK2) isoforms determines their different effect on p53 stability in tumour cell lines. AB - VRK is a new kinase family of unknown function. Endogenous human vacinia-related kinase 2 (VRK2) protein is present in both the nucleus and the cytosol, which is a consequence of alternative splicing of two VRK2 messages coding for proteins of 508 and 397 amino acids, respectively. VRK2A has a C-terminal hydrophobic region that anchors the protein to membranes in the endoplasmic reticulum (ER) and mitochondria, and it colocalizes with calreticulin, calnexin and mitotracker; whereas VRK2B is detected in both the cytoplasm and the nucleus. VRK2A is expressed in all cell types, whereas VRK2B is expressed in cell lines in which VRK1 is cytoplasmic. Both VRK2 isoforms have an identical catalytic N-terminal domain and phosphorylate p53 in vitro uniquely in Thr18. Phosphorylation of the p53 protein in response to cellular stresses results in its stabilization by modulating its binding to other proteins. However, p53 phosphorylation also occurs in the absence of stress. Only overexpression of the nuclear VRK2B isoform induces p53 stabilization by post-translational modification, largely due to Thr18 phosphorylation. VRK2B may play a role in controlling the binding specificity of the N-terminal transactivation domain of p53. Indeed, the p53 phosphorylated by VRK2B shows a reduction in ubiquitination by Mdm2 and an increase in acetylation by p300. Endogenous p53 is also phosphorylated in Thr18 by VRK2B, promoting its stabilization and transcriptional activation in A549 cells. The relative phosphorylation of Thr18 by VRK2B is similar in magnitude to that induced by taxol, which might use a different signalling pathway. In this context, VRK2B kinase might functionally replace nuclear VRK1. Therefore, these kinases might be components of a new signalling pathway that is likely to play a role in normal cell proliferation. PMID- 16704423 TI - Mutant recombinant serpins as highly specific inhibitors of human kallikrein 14. AB - The reactive center loop (RCL) of serpins plays an essential role in the inhibition mechanism acting as a substrate for their target proteases. Changes within the RCL sequence modulate the specificity and reactivity of the serpin molecule. Recently, we reported the construction of alpha1-antichymotrypsin (ACT) variants with high specificity towards human kallikrein 2 (hK2) [Cloutier SM, Kundig C, Felber LM, Fattah OM, Chagas JR, Gygi CM, Jichlinski P, Leisinger HJ & Deperthes D (2004) Eur J Biochem271, 607-613] by changing amino acids surrounding the scissile bond of the RCL and obtained specific inhibitors towards hK2. Based on this approach, we developed highly specific recombinant inhibitors of human kallikrein 14 (hK14), a protease correlated with increased aggressiveness of prostate and breast cancers. In addition to the RCL permutation with hK14 phage display-selected substrates E8 (LQRAI) and G9 (TVDYA) [Felber LM, Borgono CA, Cloutier SM, Kundig C, Kishi T, Chagas JR, Jichlinski P, Gygi CM, Leisinger HJ, Diamandis EP & Deperthes D (2005) Biol Chem386, 291-298], we studied the importance of the scaffold, serpins alpha1-antitrypsin (AAT) or ACT, to confer inhibitory specificity. All four resulting serpin variants ACT(E8), ACT(G9), AAT(E8) and AAT(G9) showed hK14 inhibitory activity and were able to form covalent complex with hK14. ACT inhibitors formed more stable complexes with hK14 than AAT variants. Whereas E8-based inhibitors demonstrated a rather relaxed specificity reacting with various proteases with trypsin-like activity including several human kallikreins, the two serpins variants containing the G9 sequence showed a very high selectivity for hK14. Such specific inhibitors might prove useful to elucidate the biological role of hK14 and/or its implication in cancer. PMID- 16704424 TI - Physical properties and surface activity of surfactant-like membranes containing the cationic and hydrophobic peptide KL4. AB - Surfactant-like membranes containing the 21-residue peptide KLLLLKLLLLKLLLLKLLLLK (KL4), have been clinically tested as a therapeutic agent for respiratory distress syndrome in premature infants. The aims of this study were to investigate the interactions between the KL4 peptide and lipid bilayers, and the role of both the lipid composition and KL4 structure on the surface adsorption activity of KL4-containing membranes. We used bilayers of three-component systems [1,2-dipalmitoyl-phosphatidylcholine/1-palmitoyl-2-oleoyl phosphatidylglycerol/palmitic acid (DPPC/POPG/PA) and DPPC/1-palmitoyl-2-oleoyl phosphatidylcholine (POPC)/PA] and binary lipid mixtures of DPPC/POPG and DPPC/PA to examine the specific interaction of KL4 with POPG and PA. We found that, at low peptide concentrations, KL4 adopted a predominantly alpha-helical secondary structure in POPG- or POPC-containing membranes, and a beta-sheet structure in DPPC/PA vesicles. As the concentration of the peptide increased, KL4 interconverted to a beta-sheet structure in DPPC/POPG/PA or DPPC/POPC/PA vesicles. Ca2+ favored alpha<-->beta interconversion. This conformational flexibility of KL4 did not influence the surface adsorption activity of KL4 containing vesicles. KL4 showed a concentration-dependent ordering effect on POPG and POPC-containing membranes, which could be linked to its surface activity. In addition, we found that the physical state of the membrane had a critical role in the surface adsorption process. Our results indicate that the most rapid surface adsorption takes place with vesicles showing well-defined solid/fluid phase co existence at temperatures below their gel to fluid phase transition temperature, such as those of DPPC/POPG/PA and DPPC/POPC/PA. In contrast, more fluid (DPPC/POPG) or excessively rigid (DPPC/PA) KL4-containing membranes fail in their ability to adsorb rapidly onto and spread at the air-water interface. PMID- 16704425 TI - Spectroscopic and DNA-binding characterization of the isolated heme-bound basic helix-loop-helix-PAS-A domain of neuronal PAS protein 2 (NPAS2), a transcription activator protein associated with circadian rhythms. AB - Neuronal PAS domain protein 2 (NPAS2) is a circadian rhythm-associated transcription factor with two heme-binding sites on two PAS domains. In the present study, we compared the optical absorption spectra, resonance Raman spectra, heme-binding kinetics and DNA-binding characteristics of the isolated fragment containing the N-terminal basic helix-loop-helix (bHLH) of the first PAS (PAS-A) domain of NPAS2 with those of the PAS-A domain alone. We found that the heme-bound bHLH-PAS-A domain mainly exists as a dimer in solution. The Soret absorption peak of the Fe(III) complex for bHLH-PAS-A (421 nm) was located at a wavelength 9 nm higher than for isolated PAS-A (412 nm). The axial ligand trans to CO in bHLH-PAS-A appears to be His, based on the resonance Raman spectra. In addition, the rate constant for heme association with apo-bHLH-PAS (3.3 x 10(7) mol(-1) x s(-1)) was more than two orders of magnitude higher than for association with apo-PAS-A (< 10(5) mol(-1) x s(-1)). These results suggest that the bHLH domain assists in stable heme binding to NPAS2. Both optical and resonance Raman spectra indicated that the Fe(II)-NO heme complex is five coordinated. Using the quartz-crystal microbalance method, we found that the bHLH PAS-A domain binds specifically to the E-box DNA sequence in the presence, but not in the absence, of heme. On the basis of these results, we discuss the mode of heme binding by bHLH-PAS-A and its potential role in regulating DNA binding. PMID- 16704426 TI - Atg8L/Apg8L is the fourth mammalian modifier of mammalian Atg8 conjugation mediated by human Atg4B, Atg7 and Atg3. AB - Murine Atg8L/Apg8L has significant homology with the other known mammalian Atg8 homologs, LC3, GABARAP and GATE-16. However, it is unclear whether murine Atg8L modification is mediated by human Atg4B, Atg7 and Atg3. Expression of Atg8L in HEK293 cells led to cleavage of its C-terminus. In vitro, the C-terminus of Atg8L was cleaved by human Atg4B, but not human Atg4A or Atg4C. Atg8L-I formed an E1 substrate intermediate with Atg7(C572S), and an E2-substrate intermediate with Atg3(C264S). A modified form of Atg8L was detected in the pelletable fraction in the presence of lysosomal protease inhibitors under nutrient-rich conditions. Cyan fluorescent protein (CFP)-Atg8L colocalized with yellow fluorescent protein (YFP)-LC3 in HeLa cells in the presence of the inhibitors. However, little accumulation of the modified form of Atg8L was observed under conditions of starvation. These results indicate that Atg8L is the fourth modifier of mammalian Atg8 conjugation. PMID- 16704427 TI - Processing, catalytic activity and crystal structures of kumamolisin-As with an engineered active site. AB - Kumamolisin-As is an acid collagenase with a subtilisin-like fold. Its active site contains a unique catalytic triad, Ser278-Glu78-Asp82, and a putative transition-state stabilizing residue, Asp164. In this study, the mutants D164N and E78H/D164N were engineered in order to replace parts of the catalytic machinery of kumamolisin-As with the residues found in the equivalent positions in subtilisin. Unlike the wild-type and D164N proenzymes, which undergo instantaneous processing to produce their 37-kDa mature forms, the expressed E78H/D164N proenzyme exists as an equilibrated mixture of the nicked and intact forms of the precursor. X-ray crystallographic structures of the mature forms of the two mutants showed that, in each of them, the catalytic Ser278 makes direct hydrogen bonds with the side chain of Asn164. In addition, His78 of the double mutant is distant from Ser278 and Asp82, and the catalytic triad no longer exists. Consistent with these structural alterations around the active site, these mutants showed only low catalytic activity (relative k(cat) at pH 4.0 1.3% for D164N and 0.0001% for E78H/D164N). pH-dependent kinetic studies showed that the single D164N substitution did not significantly alter the logk(cat) vs. pH and log(k(cat)/Km) vs. pH profiles of the enzyme. In contrast, the double mutation resulted in a dramatic switch of the logk(cat) vs. pH profile to one that was consistent with catalysis by means of the Ser278-His78 dyad and Asn164, which may also account for the observed ligation/cleavage equilibrium of the precursor of E78H/D164N. These results corroborate the mechanistic importance of the glutamate-mediated catalytic triad and oxyanion-stabilizing aspartic acid residue for low-pH peptidase activity of the enzyme. PMID- 16704430 TI - Congenital mitral stenosis presenting with haemolytic anaemia. PMID- 16704431 TI - 18F-fluorodeoxyglucose positron emission tomography/computed tomography in the evaluation of early response in a primary hepatic lymphoma. PMID- 16704432 TI - A review of the scientific and literary accomplishments of Professor R.G. Macfarlane CBE, FRS. AB - This article gives an account of some of the pioneer work of Professor R.G. Macfarlane CBE, FRS in haemostasis and fibrinolysis. His Cascade hypothesis, which together with the Waterfall, became the most cited of all papers on blood coagulation in the latter part of the 20th century, is described, together with its rebuttal and defence. Macfarlane had the gift of writing in elegant prose and his papers on the philosophy of science and his biographies of Lord Florey and Fleming are reviewed. The biography of Sir Alexander Fleming is a great work that deserves to be on the shelves of every library. PMID- 16704433 TI - The diagnosis and management of factor VIII and IX inhibitors: a guideline from the United Kingdom Haemophilia Centre Doctors Organisation. AB - The revised UKHCDO factor (F) VIII/IX Inhibitor Guidelines (2000) are presented. A schema is proposed for inhibitor surveillance, which varies according to the severity of the haemophilia and the treatment type and regimen used. The methodological and pharmacokinetic approach to inhibitor surveillance in congenital haemophilia has been updated. Factor VIII/IX genotyping of patients is recommended to identify those at increased risk. All patients who develop an inhibitor should be considered for immune tolerance induction (ITI). The decision to attempt ITI for FIX inhibitors must be carefully weighed against the relatively high risk of reactions and the nephrotic syndrome and the relatively low response rate observed in this group. The start of ITI should be deferred until the inhibitor has declined below 10 Bethesda Units/ml, where possible. ITI should continue, even in resistant patients, where it is well tolerated and so long as there is a convincing downward trend in the inhibitor titre. The choice of treatment for bleeding in inhibitor patients is dictated by the severity of the bleed, the current inhibitor titre, the previous anamnestic response to FVIII/IX, the previous clinical response and the side-effect profile of the agents available. We have reviewed novel dose-regimens and modes of administration of FEIBA (factor VIII inhibitor bypassing activity) and recombinant activated FVII (rVIIa) and the extent to which these agents may be used for prophylaxis and surgery. Bleeding in acquired haemophilia is usually treated with FEIBA or rVIIa. Immunosuppressive therapy should be initiated at the time of diagnosis with Prednisolone 1 mg/kg/d +/- cyclophosphamide. In the absence of a response to these agents within 6 weeks, second-line therapy with Rituximab, Ciclosporin A, or other multiple-modality regimens may be considered. PMID- 16704434 TI - Treatment of severe aplastic anaemia with combined immunosuppression: anti thymocyte globulin, ciclosporin and mycophenolate mofetil. AB - Severe aplastic anaemia (SAA) can be successfully treated with immunosuppressive therapies or haematopoietic stem cell transplantation (HSCT). Response rates with horse anti-thymocyte globulin (h-ATG) plus ciclosporin (CsA) are about 60-70%, and robust responders have an excellent long-term survival. We introduced a third immunosuppressive agent to standard h-ATG/CsA, mycophenolate mofetil (MMF), in an attempt to improve the response rate and survival, and to decrease the relapse rate and clonal evolution to myelodysplasia. A total of 104 consecutive patients with SAA were treated with h-ATG/CsA/MMF between May 1999 and June 2003 at the National Institutes of Health Clinical Center. The overall response rate at 6 months was 62%, with 24 (37% of responders) patients relapsing at a median of 389 d from ATG. Nine patients showed evidence of clonal evolution following ATG. After a median follow up of 42 months, the median survival among responders was not reached and among non-responders was 58 months. Over half of the relapses occurred during MMF administration. Despite a strong theoretical rationale for its use, MMF did not result in the improvement of response or relapse rates when compared with historical standard h-ATG/CsA. PMID- 16704435 TI - Transformed diffuse large B-cell lymphomas with gains of the discontinuous 12q12 14 amplicon display concurrent deregulation of CDK2, CDK4 and GADD153 genes. AB - Transformation of the indolent follicular lymphoma (FL) to the aggressive diffuse large B-cell lymphoma (DLBCL) results in resistance to therapy with shortened survival. It has been demonstrated that the 12q12-14 region was mainly amplified in DLBCL cases but not in their FL counterparts. Therefore, we examined the DNA copy number and protein expression profiles for CDK2, CDK4 and GADD153, three genes that map to 12q12-14, in a set of 44 paired FL/DLBCL samples from 22 patients. The concordant amplification of these genes occurred in seven of 22 (32%) of FL cases, compared with 15 of 22 (68%) of DLBCL cases. At the protein level, 15 of 22 of the DLBCL samples (68%) showed strong staining for the CDK2 protein, compared with five of 21 of FL samples (24%). The majority of the DLBCL samples (16/22, 72%) expressed the CDK4 protein, whereas the majority of the FL samples (12/21, 57%) showed no expression of this protein. Except for one DLBCL case, no expression of the GADD153 protein could be detected. The deregulation of the CDK2 and CDK4 genes at the genetic and protein levels suggest a functional role for these genes in the transformation process and could potentially provide targets for prognostic tests or therapeutic interventions. PMID- 16704436 TI - Retreatment with rabbit anti-thymocyte globulin and ciclosporin for patients with relapsed or refractory severe aplastic anaemia. AB - The management of patients with severe aplastic anaemia (SAA) who do not have a matched sibling donor and fail a course of horse anti-thymocyte globulin (h ATG)/ciclosporin (CsA) is uncertain. Repeated courses of ATG-based immunosuppression are often employed; in children and increasingly in adults, alternative donor haematopoietic stem cell transplantation is an option. We analysed the success rate of retreatment with rabbit ATG (r-ATG)/CsA in 43 patients treated at our institution in the last 5 years; 22 were refractory (20 adults; two children) to h-ATG/CsA-based regimens and 21 (17 adults; four children) had relapsed after h-ATG/CsA-based regimens. The overall response rate was 30% in patients who were refractory to h-ATG and 65% in patients who had relapsed following h-ATG. The 1000-d survival in patients who responded to r-ATG was 90% compared with 65% in non-responders. Six patients developed a clonal haematological disorder; two were responders, two were non-responders and in two the evolution occurred before the response could be assessed at 3 months following r-ATG. Thirteen patients died; three were responders, six were non responders and four patients died prior to 3 months when response was assessed. In our study, the response rate in refractory patients was inferior to what has been previously reported. PMID- 16704437 TI - Timing of autologous stem cell transplantation from last chemotherapy affects lymphocyte collection and survival in non-Hodgkin lymphoma. AB - Autograft absolute lymphocyte count (A-ALC) is a prognostic factor for survival in non-Hodgkin lymphoma (NHL) after autologous stem cell transplantation (ASCT). An A-ALC is dependent upon the preaphaeresis absolute lymphocyte count (PA-ALC) at the time of aphaeresis. It was hypothesised that the time interval from last chemotherapy (TILC) to aphaeresis affects PA-ALC. One hundred and sixty consecutive NHL patients who underwent ASCT at the Mayo Clinic between 1996 and 2001 were evaluated. A strong correlation between TILC and PA-ALC (r = 0.67, P < 0.0001) was identified. Higher PA-ALC was observed in TILC > or =55 d compared with TILC <55 d [median: 7.0 vs. 3.8 x 10(9)/l], P < 0.0001). TILC as a continuous variable was identified as a prognostic factor for overall survival (OS) [hazard ratio (HR) = 0.989, P < 0.01] and progression-free survival (PFS) (HR = 0.992, P < 0.0492). Median OS and PFS were longer in the TILC > or =55 d vs. TILC <55 d group (not reached vs. 21 months, P < 0.0008; 76 vs. 9 months, P < 0.0025, respectively). Multivariate analysis demonstrated TILC to be an independent prognostic indicator for OS and PFS. These findings suggest that the immune status of the host at the time of aphaeresis may predict survival after ASCT. PMID- 16704438 TI - Haematopoietic stem cell transplantation as primary therapy of sporadic adult Burkitt lymphoma. AB - High dose chemoradiotherapy and haematopoietic stem cell transplantation (SCT) is used as primary therapy for patients diagnosed with Burkitt lymphoma (BL). Forty three adults presented with sporadic BL in British Columbia between 1987 and 2003. Twenty patients had bone marrow involvement. Sixteen patients did not proceed to SCT because of chemorefractory disease (n = 9) or other reasons (n = 7). Twenty-seven patients proceeded to SCT and had a 3-year event-free survival of 51%. In conclusion, approximately 50% of patients with chemosensitive BL who undergo SCT can be cured; however, a significant number of patients will not proceed to SCT because of early resistance or recurrence. PMID- 16704439 TI - C-terminal nucleophosmin mutations are uncommon in chronic myeloid disorders. AB - C-terminal somatic mutations in nucleophosmin (NPM), a nucleolar shuttling protein that binds p53 and p19(Arf), were recently described in karyotypically normal acute myeloid leukaemia (AML). We analysed primary marrow samples from 150 patients with various chronic myeloid disorders for mutations in the NPM1 gene encoding NPM. NPM1 mutations (tetranucleotide duplication) were detected in three patients, all of whom had chronic myelomonocytic leukaemia (CMML) and a short (<1 year) survival, with rapid progression to overt AML. All other patients were NPM1 wild type in the region analysed. In conclusion, C-terminal NPM mutations are uncommon in chronic myeloid neoplasia, but if present may represent an evolving leukaemic clone. PMID- 16704440 TI - Overexpression of the VAV proto-oncogene product is associated with B-cell chronic lymphocytic leukaemia displaying loss on 13q. AB - The expression of the VAV proto-oncogene in 57 patients with chronic myeloproliferative disease (CMD), B-cell acute lymphoblastic leukaemia (B-ALL) and B-cell non-Hodgkin Lymphoma (B-NHL), and 61 with B-cell chronic lymphocytic leukaemia (B-CLL) was analysed. VAV overexpression was observed in 19.5% of cases and 81% of VAV-positive tumours also displayed VAV phosphorylation. Overexpression was not observed in B-ALL or CMD, but 13% of B-NHL and 34.4% of B CLL patients (P = 0.002) overexpressed VAV. The overexpression and phosphorylation of VAV was detected more frequently in 13q- chronic lymphocytic leukaemias (71.4%) versus other B-CLLs (23.4%, P = 0.001). Overexpression of VAV protein is a frequent event in patients with B-CLL displaying loss of 13q sequences. PMID- 16704441 TI - Long-term results of an ultra low-dose cytarabine-based regimen for the treatment of acute megakaryoblastic leukaemia in children with Down syndrome. AB - Children with acute megakaryoblastic leukaemia (AMKL) and Down syndrome (DS) show a favourable response to chemotherapy, probably due to increased sensitivity of the leukaemic blasts to cytarabine. In contrast, dose-intensive approaches have resulted in disproportionate treatment-related mortality in this group. The survival of children with AMKL and DS was retrospectively compared following treatment with a low-dose chemotherapy protocol, consisting of cytarabine (10 mg/m2/dose), retinylpalmitate and vincristine or standard chemotherapy. Event free (67 +/- 11%) and overall survival (77 +/- 10%) at 5 years were not significantly different in both groups. Further reduction of treatment intensity in AMKL of children with DS, therefore, appears feasible. PMID- 16704442 TI - Allogeneic stem cell transplantation for severe acquired aplastic anaemia using a fludarabine-based preparative regimen. AB - We reviewed our experience in the treatment of 13 patients with severe acquired aplastic anaemia, using a newly developed non-myeloablative regimen consisting of fludarabine (total dose 180 mg/m2), cyclophosphamide (total dose 120 mg/kg), and antithymocyte globulin (total dose 40 mg/kg). All except one patient received multiple transfusions and had failed prior immunosuppressive treatment. Twelve out of 13 patients achieved sustained engraftment. One patient was not evaluable for engraftment because of early death on day +10. None of the patients developed graft failure. Mucositis of mild-to-moderate severity was the only observed regimen-related toxicity. The cumulative incidence of acute graft-versus-host disease (GvHD) grade II-IV and III-IV was 8.3% and 0%, respectively. With a median follow-up period of 45 months, the 5-year overall survival probability was 84%. Eight out of 11 surviving patients have been followed for more than 1 year and only one developed limited chronic GvHD. All patients enjoy a normal life style, with a Karnofsky score of 100%, and all except three, followed for 3, 5 and 6 months respectively, are free of any immunosuppressive medication. The results of this study look promising, while prospective clinical trials may be required to confirm the benefits of this regimen as an alternative to existing protocols. PMID- 16704443 TI - Measurement of von Willebrand factor binding to a recombinant fragment of glycoprotein Ibalpha in an enzyme-linked immunosorbent assay-based method: performances in patients with type 2B von Willebrand disease. AB - Type 2B von Willebrand disease (VWD) is characterised by an increased affinity of von Willebrand factor (VWF) for its platelet receptor glycoprotein Ib (GPIb). This feature is usually studied in vitro by a ristocetin-dependent VWF platelet binding assay, which has some limitations as it requires [e.g. (radio)-labelled anti-VWF antibodies and normal formaldehyde-fixed platelets]. We, here, extended the applicability of an enzyme-linked immunosorbent assay-based method previously described for the measurement of ristocetin co-factor activity that used a recombinant fragment of GPIb (rfGPIb alpha) and horseradish peroxidase-labelled rabbit anti-human VWF antibodies for measuring the captured ristocetin-VWF complexes on the rfGPIb alpha. Thirty-one type 2B VWD patients from 15 families with eight different known mutations were studied. VWF in plasma from 28 of these patients bound better than normal VWF at 0.2 mg/ml ristocetin, with the ratio, optical density (OD) patient/OD normal pool plasma, higher than 1.8. For two of the three other patients with no enhanced response of plasma VWF, the platelet lysate VWF showed an enhanced binding capacity; for the last patient, the results in other members of the family are unequivocal. We conclude that, this new method for measurement of plasma or platelet VWF-binding capacity offers great advantages for correct type 2B VWD diagnosis. PMID- 16704444 TI - Distinguishing between type 2B and pseudo-von Willebrand disease and its clinical importance. AB - Pseudo-von Willebrand disease (p-VWD) and type 2B von Willebrand disease (VWD) have similar phenotypic parameters and clinical symptoms, but different aetiologies. Fourteen individuals from five families with a historical diagnosis of type 2B VWD but with no mutation in the von Willebrand factor gene were re investigated for the possibility of p-VWD, using platelet aggregation in the presence of cryoprecipitate. p-VWD was confirmed by targeted DNA sequencing of the glycoprotein Ibalpha gene, identifying a heterozygous Glycine 233 Valine substitution. This study suggests that p-VWD may be under diagnosed, and that platelet aggregation in the presence of cryoprecipitate is useful in differentiating this disorder from type 2B VWD. PMID- 16704445 TI - Severe infections in thalassaemic patients: prevalence and predisposing factors. AB - The incidence of infections among patients with thalassaemia and the role of risk factors for infection are uncertain. We studied the occurrence of infections necessitating hospitalisation in 92 homozygous beta-thalassaemia patients who had been followed longitudinally for decades, and investigated the role of potential risk factors for these infections. Pneumonia accounted for 26% of the infections and fever of unknown origin for 14%. Staphylococcus aureus was the major pathogen possibly related to injections associated with intensive chelation with deferoxamine. There was a significant increase in the rate of infection over time, notably after 15 years. Splenectomy correlated with the incidence of infection (P < 0.001) without being confounded by other variables and with highest frequencies of infections present after 10 years. A direct correlation between iron overload and infection was evident only before the initiation of iron-chelating treatment (P < 0.01). Following initiation of deferoxamine, paradoxically, the infection rate increased (P = 0.046). The combination of splenectomy and deferoxamine treatment was associated with the highest adjusted infection rate. Parathyroid dysfunction and glucose-6-phosphate dehydrogenase deficiency were significantly associated with infection (P = 0.02 and P = 0.04 respectively). The infection rate in thalassaemia is affected mainly by the duration of the disease and is increased by splenectomy and, in the long term, by treatment with deferoxamine. PMID- 16704446 TI - Alpha-haemoglobin stabilising protein is a quantitative trait gene that modifies the phenotype of beta-thalassaemia. AB - It has been suggested that altered levels or function of alpha-haemoglobin stabilising protein (AHSP), an erythroid-specific protein that binds specifically to free alpha-(haemo)globin, might account for some of the clinical variability in beta-thalassaemia. To assess the variation of AHSP expression, mRNA levels in circulating reticulocytes of 103 healthy individuals were measured by quantitative reverse transcription-polymerase chain reaction. AHSP expression varied up to threefold, and did not correlate with age or sex. A systematic survey of the AHSP locus identified eight sequence variants, of which six were common. Four common variants, including the longer homopolymer (T18) in the putative promoter, are strongly associated with AHSP expression. Reporter assays in K562 cells showed that the activity of the shorter (T15) reporter was relatively lower than that of the T18 reporter. In a study of nine anaemic patients who were heterozygous for beta-thalassaemia and also heterozygous for the triplicated alpha-globin gene (alpha alpha alpha/alpha alpha), frequency of the shorter homopolymer was higher than expected. AHSP expression is variable, with cis control accounting for some of its variance. In some families, the subtle altered levels in AHSP related to the AHSP genotype appears to be a relevant contributory factor in the haematological phenotype. PMID- 16704447 TI - Pyruvate kinase deficiency in France: a 3-year study reveals 27 new mutations. AB - Pyruvate kinase (PK) deficiency is the most common enzyme defect affecting the glycolytic pathway of the erythrocyte. Usually, it is clinically silent in heterozygotes but serious disorders are described at birth in homozygotes or compound heterozygotes. Including the mutants herein reported, more than 180 mutations of the PK-LR gene have now been identified. This 3-year study was carried out to detect mutations associated with disease-affecting families. Haematological indices, erythrocyte PK and glucose-6-phosphate dehydrogenase activities were measured. Molecular characterisation of the PK gene mutations included restriction enzyme analysis, mutation scanning and gene sequencing. Among the 56 families studied, nine homozygous cases and 41 different mutations were found. Eight mutations involved a splice site, 31 missense mutations were located in crucial domains of the molecule (catalytic site, cleft between the A and C domains, A/A' interface) and two cases of insertion-deletion were found. In total, 20 new mutations modifying the structure of the enzyme and seven affecting a splice site are reported. PK deficiency is an under diagnosed disease. However, deficiency could be life threatening in perinatal period and we report two lethal cases. These results support the characterisation of PK mutations, and show that prenatal diagnosis can identify affected infants and prepare safer conditions for the birth. PMID- 16704448 TI - Plasma as a source of mRNA for determining IgV(H) mutation status in patients with chronic lymphocytic leukaemia. PMID- 16704449 TI - 101 years of McCrae's (not Auer's) rods. PMID- 16704450 TI - Increased CA-15.3 levels in the serum of patients with homozygous beta thalassaemia and sickle cell/beta-thalassaemia. PMID- 16704451 TI - Editorial: history and all that. PMID- 16704452 TI - The discovery of the human melanocyte. AB - Around 2200 bc the first written description of a human pigmentation disorder, most likely vitiligo, was recorded, and from that moment the history of research into human pigmentation can be traced. For the following 4000 yr, the origins of human skin colour remained an enigma that was to generate a multitude of misconceptions. Even after European physicians began to dissect and compare dark and light coloured skin to reveal its underlying anatomy, the origins of skin and hair pigmentation were a matter of frequently erroneous speculation. The true source of human pigmentation was only finally revealed with the discovery of the melanocyte in the 19th century. Once tyrosinase was identified to be the key enzyme in pigment formation, attention focused on elucidating the chemical structure of melanin, an enterprise that remains incomplete. The developmental origins of the melanocyte were described from 1940 to 1960, and the concept of the epidermal melanin unit was introduced together with a description of the ultrastructure of the melanosome and melanosome transfer. With these advances came the realization that different skin types exhibit distinct differences at the histological level that relate to varying amounts of eumelanin and pheomelanin produced by the melanocytes. The foundation established over the past 4000 yr is the basis for all current research into this fascinating cell type. PMID- 16704453 TI - Skin layer-specific transcriptional profiles in normal and recessive yellow (Mc1re/Mc1re) mice. AB - The melanocortin 1 receptor (Mc1r) plays a central role in cutaneous biology, but is expressed at very low levels by a small fraction of cells in the skin. In humans, loss-of-function MC1R mutations cause fair skin, freckling, red hair, and increased predisposition to melanoma; in mice, Mc1r loss-of-function is responsible for the recessive yellow mutation, associated with pheomelanic hair and a decreased number of epidermal melanocytes. To better understand how Mc1r signaling affects different cutaneous phenotypes, we examined large-scale patterns of gene expression in different skin components (whole epidermal sheets, basal epidermal cells and whole skins) of neonatal (P2.5) normal and recessive yellow mice, starting with a 26K mouse cDNA microarray. From c. 17 000 genes whose levels could be accurately measured in neonatal skin, we identified 883, 2097 and 552 genes that were uniquely expressed in the suprabasal epidermis, basal epidermis and dermis, respectively; specific biologic roles could be assigned for each class. Comparison of normal and recessive yellow mice revealed 69 differentially expressed genes, of which the majority had not been previously implicated in Mc1r signaling. Surprisingly, many of the Mc1r-dependent genes are expressed in cells other than melanocytes, even though Mc1r expression in the skin is confined almost exclusively to epidermal melanocytes. These results reveal new targets for Mc1r signaling, and point to a previously unappreciated role for a Mc1r-dependent paracrine effect of melanocytes on other components of the skin. PMID- 16704454 TI - Regulation of pigmentation in zebrafish melanophores. AB - In comparison with the molecular genetics of melanogenesis in mammals, the regulation of pigmentation in poikilothermic vertebrates is poorly understood. Mammals undergo morphological colour change under hormonal control, but strikingly, many lower vertebrates display a rapid physiological colour change in response to the same hormones. The recent provision of extensive genome sequencing data from teleost zebrafish, Danio rerio, provides the opportunity to define the genes and proteins mediating this physiological pigment response and characterise their function biologically. Here, we illustrate the background adaptation process in adults and larvae and describe a novel assay to visualize and directly quantify the rate of zebrafish melanophore pigment translocation in unprecedented detail. We demonstrate the resolution of this assay system; quantifying the zebrafish melanophore response to melanin-concentrating and melanocyte-stimulating hormones. Furthermore, we investigate the intracellular signalling downstream of hormone stimulation and the biomechanical processes involved in zebrafish pigment translocation, confirming the importance of cyclic adenosine monophosphate (cAMP) as a mediator of pigment translocation and finding intact microtubules are essential for both melanin dispersion and aggregation in zebrafish, but that microfilament disruption affects aggregation only. In conclusion, we propose these data establish the zebrafish as an experimental model for studying both physiological colour change and the molecular basis of pigment translocation. PMID- 16704455 TI - Melanogenesis and evidence for melanosome transport to the plasma membrane in a CD83 teleost leukocyte cell line. AB - Visceral organs of ectothermic vertebrates harbour melanin-containing leukocytes termed melanomacrophages. These cells are thought to participate in immune reactions and free-radical trapping. In teleosts, the melanin-producing ability of melanomacrophages has hitherto not been confirmed by molecular techniques. Here, a leukocyte marker and the apparatus for melanosome production and transport were investigated in an Atlantic salmon (Salmo salar) pronephros derived mononuclear leukocyte (SHK-1) cell line. The SHK-1 cells expressed transcripts specific for a mammalian CD83 homologue, a standard surface marker for activated or differentiated dendritic cells, and dopachrome tautomerase/tyrosinase-related protein-2, a melanocyte specific enzyme essential for melanin production. Reduction potential of melanin or its precursors was demonstrated histochemically after prolonged cultivation. Ultrastructural investigations revealed tyrosinase and acid phosphate activity in identical organelles and BSA-gold co-localized with multilamellar melanosomes after 2 h internalization. Apparently, melanosomes were transported and released through periodically occurring tubules fusing with the plasma membrane. Video monitoring revealed filopodia and macropinocytosis. These results showed that the SHK-1 cell line is capable of melanogenesis and melanosome secretion. Melanin-producing cells in teleost pronephros may represent a distinct CD83(+) leukocyte population consisting of phylogenetically relict multifunctional cells. This is the first report of a melanin-producing leukocyte cell-line. PMID- 16704456 TI - A polymorphism in the agouti signalling protein (ASIP) is associated with decreased levels of mRNA. AB - To date, a role for agouti signalling protein (ASIP) in human pigmentation has not been well characterized. It is known that agouti plays a pivotal role in the pigment switch from the dark eumelanin to the light pheomelanin in the mouse. However, because humans do not have an agouti banded hair pattern, its role in human pigmentation has been questioned. We previously identified a single polymorphism in the 3'-untranslated region (UTR) of ASIP that was found at a higher frequency in African-Americans compared with other population groups. To compare allele frequencies between European-Australians and indigenous Australians, the g.8818A --> G polymorphism was genotyped. Significant differences were seen in allele frequencies between these groups (P < 0.0001) with carriage of the G allele highest in Australian Aborigines. In the Caucasian sample set a strong association was observed between the G allele and dark hair colour (P = 0.004) (odds ratio 4.6; 95% CI 1.4-15.27). The functional consequences of this polymorphism are not known but it was postulated that it might result in message instability and premature degradation of the transcript. To test this hypothesis, ASIP mRNA levels were quantified in melanocytes carrying the variant and non-variant alleles. Using quantitative real-time polymerase chain reaction the mean ASIP mRNA ratio of the AA genotype to the AG genotype was 12 (P < 0.05). This study suggests that the 3'-UTR polymorphism results in decreased levels of ASIP and therefore less pheomelanin production. PMID- 16704457 TI - pyewacket, a new zebrafish fin pigment pattern mutant. AB - Many mutants that disrupt zebrafish embryonic pigment pattern have been isolated, and subsequent cloning of the mutated genes causing these phenotypes has contributed to our understanding of pigment cell development. However, few mutants have been identified that specifically affect development of the adult pigment pattern. Through a mutant screen for adult pigment pattern phenotypes, we identified pyewacket (pye), a novel zebrafish mutant in which development of the adult caudal fin pigment pattern is aberrant. Specifically, pye mutants have fin melanocyte pigment pattern defects and fewer xanthophores than wild-type fins. We mapped pye to an interval where a single gene, the zebrafish ortholog of the human gene DHRSX, is present. pye will be an informative mutant for understanding how xanthophores and melanocytes interact to form the pigment pattern of the adult zebrafish fin. PMID- 16704458 TI - Oculocutaneous albinism with TYRP1 gene mutations in a Caucasian patient. AB - Non-syndromic oculocutaneous albinism (OCA) is a clinically and genetically heterogeneous autosomal recessive disorder with mutations identified in several genes: OCA1 (tyrosinase, TYR), OCA2 (OCA2), OCA3 (tyrosinase-related protein 1, TYRP1), and OCA4 (membrane-associated transporter protein, MATP). OCA3 was thought to be restricted to black populations, where it was clinically described as rufous or brown albinism, until the recent report of a homozygous TYRP1 mutation in Caucasian patients from a consanguineous Pakistani family. Here, we describe a German patient of Caucasian origin, with a light-yellow skin, yellow gold hair with orange highlights, fair eyelashes, several pigmented naevi, and no tendency to tan, only to burn. Eye-colour is blue-green with substance defects of the iris. Molecular analysis did not reveal any mutation in the TYR and OCA2 genes. Two mutations were found in the TYRP1 gene: a missense mutation (c.1066G>A/p.Arg356Glu) that was inherited from the mother, and a de novo single base deletion (c.106delT/p.Leu36X). This finding suggests that mutation screening should be extended to the TYRP1 gene in patients from all ethnic origins, at least in cases where no mutations have been identified in the other OCA genes. PMID- 16704459 TI - Color reversion of the albino medaka fish associated with spontaneous somatic excision of the Tol-1 transposable element from the tyrosinase gene. AB - The medaka fish albino mutant, i(1) is one of the Tomita collection of medaka pigmentation mutants which exhibits a complete albino phenotype, because of inactivation of the tyrosinase gene due to insertion of a transposable element, Tol-1. Recently, mosaic black-pigmented i(1) medaka fish have arisen in one of our laboratory breeding populations. Their pigmented cells have been observed in all of the tissues, including the eye and skin, in which melanin is detectable in the wild type. In this study, we analyzed the tyrosinase gene of revertants and showed Tol-1 to have been precisely excised from the gene, suggesting a causal relationship. Mosaic patterns of pigmentation indicate spontaneous somatic excision of the element from the tyrosinase gene. To our knowledge, this is the first transposable element with somatic excision activity demonstrated phenotypically in vertebrates. The pattern of pigmentation in mosaic revertants indicates frequencies of melanin pigments to be consistent with the numbers of melanophores per unit area of body sites, such as the eyes, head and dorsal trunk. PMID- 16704460 TI - Characterization of the pigment produced by the planarian, Dugesia ryukyuensis. PMID- 16704461 TI - Pmel17: controversial indeed but critical to melanocyte function. PMID- 16704464 TI - The parasite immunologist's crystal ball. PMID- 16704465 TI - Crystal ball. PMID- 16704468 TI - Human papillomavirus infection: an old disease, a new vaccine. AB - Human papillomavirus (HPV) infection is common and causes a wide spectrum of disease. With recent advances in the development of prophylactic HPV vaccines, it is likely that these will be licensed for use in the near future. This review focuses on the science behind HPV vaccines, published clinical trial results for both prophylactic and therapeutic HPV vaccines, important issues relevant to implementation and cost-effectiveness models of HPV vaccination programs. It may be that an HPV vaccine that protects against the complications of HPV infection such as cervical cancer will be one of the most significant public health initiatives of this decade. PMID- 16704469 TI - Refractive issues in pregnancy. AB - Pregnancy is known to cause refractive changes as a result of various hormonal changes occurring during pregnancy. These changes may persist for a few weeks post-partum and during lactation. In this article we discuss various refractive issues in pregnancy that have an effect on contact lens use, myopia and the outcome of refractive surgery. PMID- 16704470 TI - Preterm outcome table (POT): a simple tool to aid counselling parents of very preterm infants. AB - BACKGROUND: Outcome figures published in scientific journals are often cumbersome and difficult to understand by parents during counselling before or immediately after a very premature birth. AIM: To provide simplified up-to-date outcome information in a table for ease of counselling. METHODS: Regional perinatal mortality rates for very premature births (23-31 weeks gestation) and incidence of significant neonatal events for those admitted to neonatal intensive care units (NICU) were obtained from the NSW Midwives Data Collection, ACT Maternal and Perinatal Data Collection and the NSW and ACT NICUS Data Collection for 2000 and 2001. Neurodevelopmental outcome was obtained for the same cohort at 2-3 years of age, corrected for prematurity. The percentage outcomes were rounded off to the closest conservative multiple of 5 for each data point in a table. RESULTS: The preterm outcome table (POT) for each gestational week was constructed from a total of 2315 births. Of these, 401 (17.3%) were reported as stillborn and were predominantly of 23 to 25 weeks gestation. Of those admitted to NICU, hospital survival rates were 30, 50, 65, 75, 80, 90 and > 95% for 23, 24, 25, 26, 27, 28-29 and 30-31 weeks, respectively. Neurodevelopmental outcome was available for 470 (75%) children, of whom 15% had a moderate to severe functional disability at 2-3 years of age, corrected for prematurity. Simplified data on survival to discharge and outcome were tabulated. CONCLUSION: POT appears simple and easy to use but also provides realistic data to assist clinicians in the counselling process. PMID- 16704471 TI - First trimester medical termination of pregnancy: an alternative for New Zealand women. AB - BACKGROUND: First trimester medical terminations of pregnancy (TOP) have been carried out in a private abortion clinic in Auckland from 1 July 2003 to 30 June 2005. AIMS: To audit first trimester medical TOP outcomes over the first 2 years and to compare the demographics and complication rates of women opting for medical TOP with those choosing surgical TOP during this period. METHODS: Retrospective, consecutive case audit of women presenting for a first trimester TOP. Anonymous audit included ethnicity, gestation at first appointment, pain relief requirements, blood loss, timing from the taking of misoprostol to expulsion of products of conception, complications and comparison of outcomes and characteristics between medical termination of pregnancy (MTOP) and surgical termination of pregnancy (STOP). RESULTS: A total of 3311 TOPs were performed over the 2-year study period, including 390 MTOPs (12%). No significant differences were found in age, ethnicity or history of previous TOP between women choosing MTOP or STOP. Gravida 1 and nulliparous women were significantly more likely to choose MTOP. Five per cent (21/390) of women taking mifepristone progressed to STOP. Complications involving hospitalisation occurred in six MTOPs (1.5%) and 18 STOPs (0.6%). CONCLUSIONS: First trimester MTOP is safe and effective and is a realistic alternative for women wanting choice of method in New Zealand. PMID- 16704472 TI - Compliance with a risk factor-based intrapartum prophylaxis program for neonatal group B streptococcal disease. AB - AIM: The US Centre for Disease Control (CDC) recently amended their guidelines for the prevention of early-onset group B streptococcal disease (EOGBSD) of the newborn to recommend bacteriological screening, rather than risk factor-based screening, as the preferred method of identifying 'at risk' mother-infant pairs. This recommendation was derived from population data suggesting that the effectiveness of bacteriological screening was superior to a risk-factor approach because antibiotic compliance was better with the former. Whether poor compliance and therefore impaired prevention is inherent in risk-factor screening has not been widely tested. METHODS: For a 6-month period we audited compliance with an established risk-factor EOGBSD prophylaxis program. RESULTS: During the audit period, 1243 women delivered, of whom 287 (23%) had at least one risk factor. Of these women, 193 (67%), representing 15% of all women giving birth, received antibiotics. Thus, there were 94 women who were eligible for antibiotics but did not receive prophylaxis. There were sound clinical reasons for withholding antibiotics in 68 of these. Therefore, the corrected compliance rate within our program was 73%. CONCLUSION: This compares favourably with published compliance rates with bacteriological-based programs, but we have suggested mechanisms to improve compliance further. PMID- 16704473 TI - Recent impact of anal sphincter injury on overall Caesarean section incidence. AB - INTRODUCTION: Because of increasing recognition of obstetric anal sphincter injury and faecal incontinence, we examined the recent impact of these indications on our institutional Caesarean section incidence. METHODS: Retrospective review of the indications for multiparous Caesarean section was performed at the National Maternity Hospital for the 4 years 2000-2003, inclusive, to identify women in whom previous anal sphincter injury was an indication. Individual charts were reviewed and data regarding the nature and extent of previous anal sphincter injury were obtained. RESULTS: Among 17 586 consecutive multiparous deliveries, previous anal sphincter trauma constituted the indication for Caesarean delivery in 67 women, representing 0.4% of all multiparae, 2.9% of multiparous Caesarean sections and 1.3% of all Caesarean sections performed. Fifty (85%) of the 67 women who opted for prelabour Caesarean delivery following previous obstetric anal sphincter injury had symptoms of faecal incontinence (mean continence score 5, range 1-17). CONCLUSION: Notwithstanding recent increased awareness and documentation, anal sphincter problems represent a small influence on total Caesarean incidence. PMID- 16704474 TI - Women's and health professionals' preferences for prenatal testing for Down syndrome in Australia. AB - AIM: To describe and compare women's and health professionals' preferences for prenatal screening tests for Down syndrome. DESIGN: Cross-sectional questionnaire survey. PARTICIPANTS AND SETTING: Women (n = 322) attending for a glucose challenge test at 26 weeks gestation and health professionals (266 midwives and 34 obstetricians) at the Royal Women's Hospital, Melbourne, between 13 December 2002 and 30 April 2003. OUTCOME MEASURES: The relative value participants attach to attributes of Down syndrome screening tests as determined by conjoint analysis and ranking scales. RESULTS: Women and health professionals shared similar relative values regarding the importance of detection rate of screening tests, according to coefficients from conjoint analysis models. However, health professionals placed higher relative values on timing of prenatal tests and risk associated with the subsequent diagnostic test than did women. Comparison of coefficients suggests that, compared with health professionals, women would wait longer and accept a greater decrease in detection rate for a test if it was safer. Using the more traditional ranking scale, the safest test was ranked first by 56% of women while 47% of health professionals ranked a test with the highest detection rate first. Equal proportions ( approximately 10%) in both groups ranked the earliest test first. CONCLUSION: There is a general agreement between pregnant women and health professionals regarding the relative importance they attach to different attributes of a test. However, health professionals appeared to favour earlier timing of tests while women placed greater emphasis on safety. Utilising two different measures of preference demonstrated the complexity of decision-making. PMID- 16704475 TI - Intrauterine growth restriction and the proliferation of smooth muscle cells in umbilical vessels. AB - AIMS: The purpose of this study was to investigate the proliferation of smooth muscle cells in umbilical vessels of fetuses affected by intrauterine growth restriction (IUGR) and to compare the findings with gestational age-matched control cases. METHODS: Sixty umbilical cords from fetuses at 36-37 weeks gestation were examined. Fetuses were divided into three groups: group I, appropriate for dates birthweight; group II, IUGR with reassuring fetal status; and group III, IUGR with abnormal umbilical Doppler waveforms. Umbilical cords were immunostained with an antibody to proliferating cell nuclear antigen and Ki 67; stained smooth muscle cells were subsequently counted. Smooth muscle cell density was determined by counting the total number of cells in a representative area of vessel wall and the wall thickness of each vessel was also measured. RESULTS: Proliferation marker-positive cells were increased in the umbilical vessels of group II compared to group I, and there were more proliferating smooth muscle cells in the umbilical vessels of group III compared to the other two groups. The umbilical vessels of group III showed the highest smooth muscle cell density, but the wall thickness of all vessels was significantly thinner in group III than the other two groups. CONCLUSIONS: This study showed overproliferation of smooth muscle cells in the umbilical vessel walls associated with IUGR. It is hypothesised that hypoxia might induce this overproliferation given the further proliferation in IUGR fetuses with abnormal umbilical Doppler waveforms. Coexistence of a high cell density and lean vessel walls suggests small smooth muscle cells in umbilical vessels with IUGR. PMID- 16704476 TI - Prevalence of sexually transmitted infections in pregnant urban Aboriginal and Torres Strait Islander women in northern Australia. AB - OBJECTIVE: To assess the prevalence of sexually transmitted infections (STI) in a cohort of pregnant urban Indigenous women and association of STI with preterm birth, low birthweight birth and perinatal mortality. DESIGN: Prospective intervention program in a cohort of women attending Townsville Aboriginal and Islander Health Services (TAIHS) for shared antenatal care between 1 January 2000 and 31 December 2003 incorporating routine screening for chlamydia, gonorrhoea, trichomoniasis, hepatitis B and syphilis. SETTING: Townsville is a provincial urban centre with a regional Indigenous population of over 16 000. PARTICIPANTS: Four hundred and fifty-six pregnant women who were screened for bacterial STI and other viral infections. MAIN OUTCOME MEASURES: Prevalence of STI, associated risk factors and perinatal outcomes. RESULTS: Of the 456 women, 403 (88.4%) were screened for chlamydia, gonorrhoea and trichomonas and 432 (94.7%) were screened for syphilis. A total of 92 cases of STI (20.2%, 95% CI 16.5-23.9) were detected, with 21 concurrent infection(s). The overall prevalence of chlamydia was 14.4%, gonorrhoea 6.1%, trichomoniasis 7.2% and infectious syphilis 2.5%. Predictors for STI were young age, harmful/hazardous alcohol use and unwanted pregnancy. Low birthweight and perinatal death were significantly associated with the presence of STI and infectious syphilis during pregnancy. CONCLUSION: The prevalence of STI among pregnant women in this urban Indigenous community is high, suggesting that screening for STI should be included in all antenatal care protocols for Indigenous women in Australia. Strategies to reach the whole Indigenous community of child-bearing age, especially those aged less than 25 years, are needed to improve perinatal outcome. PMID- 16704477 TI - Ultrasound localisation and removal of non-palpable Implanon implants. AB - BACKGROUND: Implanon is the trade name for a contraceptive implant that releases etonogestrel over a 3-year period. The rod is inserted into the subdermis of the upper arm. The implant should be palpable and easily removed through a small incision. AIMS: To determine whether ultrasound is an accurate means of localising and guiding removal of non-palpable Implanon implants. METHODS: All patients referred to The Northern Hospital, between March 2002 and October 2005, for localisation and/or removal of their non-palpable Implanon implants were included in this study. Ultrasound was used to locate the implants, the implant was considered localised when a linear echogenic rod with posterior acoustic shadowing was identified. When removal of the implant was requested and the implant was clear of neurovascular structures, blunt dissection was performed under ultrasound control and the implant was extracted. If the implant was considered too close to major neurovascular structures, the position of the implant was marked and the patient was referred for surgical removal. RESULTS: Thirty-four women were included in the study. In 33 patients the implant was identified. In one patient no implant was found and its absence was confirmed by serum assay. Twenty-four patients were referred for implant removal. In 20 patients the implant was successfully removed under ultrasound guidance. In the other four patients the implant was close to neurovascular structures. The position of the implants was marked and surgical removal was successfully performed. CONCLUSION: Ultrasound is highly accurate at localising and guiding removal of non-palpable Implanon implants. PMID- 16704478 TI - A retrospective review of self-reported alcohol intake among women attending for antenatal care in Far North Queensland. AB - BACKGROUND: Fetal alcohol syndrome and fetal alcohol spectrum disorder are known to occur among children delivered to women in Far North Queensland; to date, the problem has not been quantified or related directly to maternal alcohol consumption during pregnancy. AIMS: To identify the characteristics of women who consume alcohol during pregnancy, in order to identify 'high-risk' women to whom educational intervention regarding fetal alcohol syndrome may be targeted. METHODS: A retrospective descriptive study using the case notes of all women attending the antenatal booking clinic in Cairns Base Hospital over the 3-month period from April to June 2005. Obstetric and demographic information and self reported alcohol intake before and during pregnancy were noted and self-reported alcohol intake before and during pregnancy. RESULTS: Five hundred and forty-one women were included in this study. Of these women, 24.6% were found to consume alcohol during pregnancy and 57.7% were found to consume alcohol before pregnancy, indicating a reduction in alcohol consumed once women became aware of pregnancy. The most significant indicator for alcohol consumption during pregnancy was found to be alcohol consumption before pregnancy. Of women who consumed alcohol in pregnancy, 71.5% drank moderate amounts of alcohol before pregnancy and 28.7% drank heavily before pregnancy. CONCLUSION: Alcohol consumption during pregnancy is common in Far North Queensland. Methods currently used during antenatal booking visits to determine alcohol consumption before and during pregnancy are unreliable. Strategies to identify women consuming alcohol before and during pregnancy and education programs are vitally important if the incidence of fetal alcohol syndrome and spectrum disorder is to be reduced. PMID- 16704479 TI - Clinicians' evaluations of fetal oximetry sensor placement in a multicentre randomised trial (the FOREMOST trial). AB - BACKGROUND: Fetal pulse oximetry (FPO) may improve the assessment of the fetal well-being in labour. Reports of health-care provider's evaluations of new technology are important in the overall evaluation of that technology. AIMS: To determine doctors' and midwives' perceptions of their experience placing FPO sensors. METHODS: We surveyed clinicians (midwives and doctors) following placement of a FPO sensor during the FOREMOST trial (multicentre randomised trial of fetal pulse oximetry). Clinicians rated ease of sensor placement (poor, fair, good and excellent). Potential influences on ease of sensor placement (staff category, prior experience in Birth Suite, prior experience in placing sensors, epidural analgesia, cervical dilatation and fetal station) were examined by ordinal regression. RESULTS: There were 281 surveys returned for the 294 sensor placement attempts (response rate 96%). Sensors were placed by midwives (29%), research midwives (48%), registrars (22%) and obstetricians (1%). The majority of clinicians had 1 or more years' Birth Suite experience, had placed six or more sensors previously, and rated ease of sensor placement as good. Advancing fetal station (P < 0.001) and the presence of epidural analgesia prior to sensor placement (P = 0.029) predicted improved ease of sensor placement. Having a clinician placing a sensor for the first time predicted a lower rating for ease of sensor placement (P = 0.001), compared to having placed one or more sensors previously. CONCLUSIONS: Clinicians with varying levels of Birth Suite experience successfully placed fetal oxygen saturation sensors, with the majority rating ease of sensor placement as good. PMID- 16704480 TI - Improved management in threatened preterm labour with rapid fetal fibronectin testing. AB - Recently, a new rapid system for fetal fibronectin (fFN) analysis (TLI(IQ)) has been developed with accuracy equal to the gold standard, ELISA (enzyme-linked immunosorbent assay) testing. The objective of this quality improvement project was to confirm that a reduction in intervention and a reduced length of stay did occur, as expected, after the testing of fFN was introduced at a tertiary referral hospital (National Women's Hospital) for women in threatened preterm labour. There was a reduction in investigation and treatment of women after fFN was introduced. The average length of stay reduced from 3.8 to 2.9 days after testing was introduced. For the women who were fFN negative, the average length of stay was 1.7 days. PMID- 16704481 TI - The genetic basis of premature ovarian failure. AB - Premature ovarian failure (POF) is a common condition, affecting approximately 1:100 women. It is characterised by amenorrhea, hypoestrogenism, and elevated gonadotrophin levels in women under the age of 40. It is often an unexpected and distressing diagnosis, which coincides with infertility and menopausal symptoms. There is a well recognised genetic basis to the development of POF. Our laboratory has identified several candidate genes associated with POF. PMID- 16704482 TI - The role of activins during decidualisation of human endometrium. AB - Decidualisation of the endometrial stroma is critical to create a specialised environment for embryo implantation and trophoblast invasion; however, the mechanisms involved are poorly understood. We have established that activin A is an important regulator of decidualisation of endometrial stromal cells in vitro. Here we describe studies that verify the physiological significance of these findings. We demonstrate that high concentrations of activin A are produced by decidualising cells in excess of the antagonists, inhibin and follistatin, thus confirming its bioavailability within the decidual environment. Furthermore, we demonstrate that all components of the activin signalling pathway (activin receptors and Smads) are expressed in decidualised cells, and identify a downstream mechanism for activin in the endometrium, through the regulation of matrix metalloproteinases (MMPs). This new knowledge is important for understanding the roles for activins and inhibins in regulating fertility. PMID- 16704483 TI - Identification and characterisation of human endometrial stem/progenitor cells. AB - The human endometrium is a dynamic, cyclically regenerating tissue. Because adult stem cells are present in other regenerative tissues, a clonal analysis of purified endometrial epithelial and stromal cells derived from hysterectomy tissue was undertaken, as a first attempt to identify and characterise endometrial stem/progenitor cells. Rare populations of epithelial and stromal cells were clonogenic. Only those cells initiating large clones, 0.09% of epithelial cells and 0.02% of stromal cells, are likely to be endometrial stem/progenitor cells. PMID- 16704484 TI - Male-to-female transsexualism: laparoscopic pelvic floor repair of prolapsed neovagina. AB - The incidence of prolapse of the neovagina after male-to-female gender reassignment surgery is unknown. We present the first case of laparoscopic total pelvic floor repair in a male-to-female transsexual. This surgical procedure combined an understanding of the anatomy of the male pelvis with the principles of laparoscopic pelvic floor repair in the XX female. PMID- 16704485 TI - Alpha-methyldopa-induced acute hepatitis in pregnancy. PMID- 16704486 TI - Invasive Haemophilus parainfluenzae maternal-infant infections: an Australasian perspective and case report. AB - Maternal and neonatal infection with Haemophilus parainfluenzae is an unusual but potentially life-threatening occurrence. We present a case of H. parainfluenzae complicating pregnancy in the second trimester. Predisposing factors for infection with Haemophilus species and Australasian data on invasive neonatal Haemophilus infections are presented. PMID- 16704487 TI - Re: Abortion politics and the impact on reproductive health care. PMID- 16704488 TI - Re: Should methotrexate be abandoned in the management of placentas left in situ in advanced abdominal pregnancies? PMID- 16704489 TI - Re: Collaboration in maternity care. A response to 'what do I think of midwife led units?'. PMID- 16704491 TI - Extrarenal rhabdoid tumors of soft tissue: clinicopathological and molecular genetic review and distinction from other soft-tissue sarcomas with rhabdoid features. AB - Malignant rhabdoid tumor (MRT) of the soft tissue is a rare and highly aggressive tumor that occurs in infancy or childhood. It predominantly involves a deep axial location such as the neck or paraspinal region. Microscopically, the tumor is composed of a diffuse proliferation of rounded or polygonal cells with eccentric nuclei, prominent nucleoli and glassy eosinophilic cytoplasm containing hyaline like inclusion bodies, arranged in sheets and nests. These characteristic 'rhabdoid cells' are also present in certain soft-tissue sarcomas such as synovial sarcoma, extraskeletal myxoid chondrosarcoma and leiomyosarcoma. The existence of rhabdoid cells in these other sarcomas is correlated with a worse prognosis for the patients. Cytogenetic and molecular analyses have shown abnormalities in the long arm of chromosome 22 and alteration of the hSNF5/INI1 (SMARCB1) gene in renal, extrarenal and intracranial MRT. This gene alteration has been considered to be a specific molecular event in MRT, but a recent study has also demonstrated frequent alteration of this gene in proximal-type epithelioid sarcoma (ES). Both MRT of soft tissue and proximal-type ES show immunoreactivity for vimentin, cytokeratin and epithelial membrane antigen. The tumor cells of proximal-type ES are also occasionally positive for CD34 and beta catenin, whereas MRT of soft tissue has no immunoreaction for these markers. Detailed clinicopathological and immunohistochemical evaluations are necessary to distinguish MRT of soft tissue from proximal-type ES, because these tumors demonstrated a similar morphology and the same gene alteration. PMID- 16704492 TI - Immunohistochemical expression of aminopeptidase N (CD13) in human lung squamous cell carcinomas, with special reference to Bestatin adjuvant therapy. AB - Bestatin, a specific inhibitor of aminopeptidase N (CD13), has been reported to prolong survival time in patients with completely resected stage I lung squamous cell carcinoma. Considering the antitumor mechanism of Bestatin, it is interesting to know whether CD13 is expressed in human lung squamous cell carcinoma. The immunohistochemical expression of CD13 was examined in human lung carcinoma and the question of whether CD13 was immunohistochemically expressed in the interstitial tissue was investigated, mainly in the fibroblasts and blood vessels, surrounding the tumor nests of various kinds of non-small cell lung cancers, especially of squamous cell carcinomas. In Japanese squamous cell carcinoma of the lung, 38 (61.3%) out of 62 cancers were positively stained in the same manner on immunohistochemistry for CD13. The area of interstitial tissue positively stained for CD13 varied depending on the case. To confirm the cell nature of the interstitial tissue with CD13 positivity, double immunohistochemistry using CD34 and alpha-smooth muscle actin was performed. Double immunohistochemistry showed that the majority of CD13-positive cells were slender fibroblastic cells around the blood vessels and some endothelial cells. PMID- 16704493 TI - Human papillomavirus localization in cervical adenocarcinoma and adenosquamous carcinoma using in situ polymerase chain reaction: review of the literature of human papillomavirus detection in these carcinomas. AB - Many studies have suggested that human papillomavirus (HPV) infection plays an important role in the carcinogenesis of the cervical adenocarcinoma. However, the prevalence of HPV infection in cervical adenocarcinoma and adenosquamous carcinoma varies among the studies. Cervical adenocarcinoma (24 cases) and adenosquamous carcinoma (16 cases), including the underlying non-neoplastic epithelium were examined for HPV-DNA using in situ polymerase chain reaction (PCR), which enabled visualization of the localization on a glass slide. In adenocarcinoma, HPV-DNA was found in 13 cases (54%) and in eight cases in underlying non-neoplastic epithelium, resulting in a total of 21 positive cases (88%). In adenosquamous carcinoma, HPV-DNA was detected in 12 cases (75%) and and the HPV-DNA localization of each component was pure adenocarcinoma, 28.6%; mixed, 54.5%; and pure squamous cell carcinoma, 83.3%. In the underlying non-neoplastic epithelium, HPV-DNA was found more frequently in the squamous epithelium (73.3%) than the cervical glands (6.3%). In conclusion, HPV-DNA was detected in 54% of adenocarcinoma, and the rate was elevated by HPV localization in the underlying non-neoplastic epithelium. HPV infection in the underlying squamous epithelium might be related to the carcinogenesis, even in cervical adenocarcinoma. HPV-DNA localization was different in each component of adenosquamous carcinoma. PMID- 16704494 TI - Prognostic factors of phyllodes tumor of the breast. AB - The phyllodes tumor is characterized by its tendency to recur locally and occasionally to metastasize. The purpose of the present paper was to assess the prognostic value of clinical-morphological characteristics in patients with phyllodes tumor. Forty-seven cases of phyllodes tumors was studied; the World Health Organization classification was used and follow up was obtained. A total of 51%, 28% and 21% of the tumors were classified as benign, borderline and malignant, respectively. The adherence (P = 0.01), size >10 cm (P = 0.001), high mitotic activity (P = 0.03), infiltrative tumor margin (P = 0.0002) and type of surgery in malignant tumors (P = 0.02) proved to be good predictors of relapse. The presence of pain (P = 0.03), postmenopausal status (P < 0.04), heavy cellular pleomorphism (P = 0.007), high mitotic activity (P = 0.002), tumoral grade (P = 0.006) and metastasis (P < 0.00001) were prognostic factors of poor survival. Tumoral grade and some clinical-morphological characteristics of patients with phyllodes tumors have a significant impact on the prediction of its biological behavior. PMID- 16704495 TI - Standard organ weights among elderly Japanese who died in hospital, including 50 centenarians. AB - The present study was conducted to determine standard organ weights among the elderly, because little has been reported on this subject. To analyze the human aging process in terms of organ weights, age-associated changes and correlations among organ weights and the contributions of age and nutrition to organ weights were also examined. The subjects included 1615 consecutive autopsy cases of patients aged 60-99 years who died between 1995 and 2003, and cases of 50 centenarians who died between 1973 and 2005. The weights of nine organs were measured before formalin fixation. If affected by serious diseases, such as cancer, the organs were excluded from the study. Values beyond 99% of the bilateral measurement limits were also excluded. In this manner the standard organ weights were obtained. The organ weights decreased significantly according to age in all organs except for the heart in men and the lungs in both genders. Undernutrition strongly contributed to organ weight except for the hypophysis, and was especially apparent in the heart and liver. In conclusion, the standard organ weights of elderly patients who died while hospitalized were determined. Undernutrition contributed significantly to a reduction in organ weights. PMID- 16704496 TI - Immunohistochemical study of immunoglobulin light chain amyloidosis with antibodies to the immunoglobulin light chain variable region. AB - To detect immunoglobulin (Ig) light chain amyloidosis (AL amyloidosis) in formalin-fixed, paraffin-embedded tissue sections by immunohistochemistry, polyclonal antibodies were generated against synthetic peptides corresponding to amino acids 1-19 of the Ig lambda light chain V lambda VI subgroup (anti-V lambda VI (1-19)) and the Ig kappa light chain Vkappa I subgroup (anti-Vkappa I (1-19)). Anti-V lambda VI (1-19) antibody reacted with amyloid deposits in 21 of 22 Alambda amyloidosis cases, and anti-Vkappa I (1-19) antibody reacted with amyloid deposits in 10 of 11 Akappa amyloidosis cases. Immunoreactivity varied in intensity by case and within specimens. Surprisingly, amyloid deposits were positive for anti-V kappa I (1-19) staining in one case of Alambda amyloidosis. Analysis of anti-V lambda VI (1-19) and anti-Vkappa I (1-19) antibody reactivity by ELISA showed some cross-reactivity with peptides other than antigen peptides. The antibodies were not reactive in all cases of AL amyloidosis examined but may be useful, together with anti-Ig constant region antibodies, for immunohistochemical diagnosis of AL amyloidosis. PMID- 16704497 TI - Spontaneous regression of intraoral mucosa-associated lymphoid tissue lymphoma: molecular study of a case. AB - Mucosa-associated lymphoid tissue (MALT) lymphoma presentation in the oral cavity is very rare. Reported herein is a case of intraoral MALT lymphoma of the minor salivary gland in a 70-year-old woman with Sjogren's syndrome. Unexpectedly, a spontaneous clinically and histologically confirmed regression occurred 1 month after the tumor biopsy for diagnosis. Considering that salivary MALT lymphoma is associated with Sjogren's syndrome and that the chronic inflammation caused by Sjogren's syndrome persisted, it is hypothesized that the tumor clone might be present in the regressed lesion. Minimal residual tumor clone identical with the primary lesion was detected using the polymerase chain reaction (PCR) clonality assay for immunoglobulin heavy chain gene (IgH) rearrangement. No recurrence was clinically evident 38 months after the diagnosis. Spontaneous regression of MALT lymphoma should be examined at the molecular level in addition to clinical and histological evaluations. When minimal residual disease is detected, close follow up is necessary for early detection of the tumor relapse. PMID- 16704498 TI - Unusual subcutaneous splenosis occurring in a gunshot wound scar: pathology and immunohistochemical identification. AB - Splenosis usually occurs after traumatic rupture of the spleen with autotransplantation of splenic tissue to ectopic sites. Most commonly, it occurs as intraperitoneal nodules, which are found either incidentally or after symptomatic complications. Subcutaneous splenosis is a rare condition mostly observed in abdominal surgical scars. Reported herein is a case of subcutaneous splenosis developed in an exit gunshot wound scar on the left lower chest wall 9 years after splenectomy. The lesion presented as an asymptomatic subcutaneous nodule, which was excised under the impression of a skin tumor. Microscopically, there were multiple subcutaneous nodules resembling splenic tissue with red and white pulp. They were confirmed to be ectopic splenic tissue by immunohistochemical staining of the vascular lining cells (CD8+, CD31+, and CD34 ). Only 11 cases of subcutaneous splenosis have been reported before. The present case was the third case occurring in an exit gunshot wound scar, and the diagnosis was confirmed by immunohistochemical study. PMID- 16704499 TI - Ciliated hepatic cyst without smooth muscle layer: a variant of ciliated hepatic foregut cyst? AB - The ciliated hepatic foregut cyst (CHFC) is a rare lesion that may arise from remnants of the embryonic foregut. Its wall is lined by pseudostratified ciliated columnar epithelium, and is characterized by the presence of bundles of smooth muscle. Herein is presented two cases of ciliated hepatic cyst without smooth muscle layer. One case was an incidental autopsy finding, and the other was a surgically resected cystic lesion of the liver. Both cysts were <2.0 cm in diameter, and were located subcapusularly in the medial segment (S4) of the liver. The histological appearance of the two cases was identical. They had an epithelial lining of ciliated pseudostratified cells with occasional goblet cells, and lacked a smooth muscle layer, as shown by the actin immunostaining. The lining epithelium contained cells positive for immunohistochemical staining of surfactant apoprotein A, suggesting the embryonic foregut origin of the cysts and differentiation toward bronchiolar structures, rather than ciliated metaplasia of the epithelium of the simple (cholangiogenic) cyst. It is considered that ciliated hepatic cysts of the present case are a rare histological variant of CHFC where the smooth muscle layer is inconspicuous or absent. PMID- 16704500 TI - Small bowel strangulation caused by parasitic peritoneal strand. AB - A 51-year-old woman with a history of eating raw fish over a period of 2 weeks developed a progressive abdominal pain and leukocytosis with signs of small bowel obstruction. Eosinophilia was not detected in the peripheral blood. The patient underwent surgery to clarify the possibility of ileus. Partial small bowel near the ileum was trapped by a peritoneal strand and was strangulated for approximately 30 cm in length with congestion and edema. The removal of the strand easily released the strangulation and the small bowel returned to a normal appearance. Pathologically, the strand consisted of granulomatous inflammation with a wide zone of necrosis containing a dead ghost feature of a parasite in its center. The necrosis was surrounded by palisading spindle cells with largely lymphocytic infiltration and a few eosinophils. In the parasite, there presented the polymyalian type muscle layer, ventricles and Renette cells, which suggested that the parasite is compatible with the third stage larva of Anisakis type I, leading to the diagnosis of ectopic anisakidosis. It should be noted that, on rare occasions, Anisakis larva migrans can form a peritoneal strand with a wide zone of necrosis and cause strangulation ileus, especially in populations with the habit of eating raw fish. PMID- 16704501 TI - Pulmonary capillary hemangiomatosis incidentally detected in a lobectomy specimen for a metastatic colon cancer. AB - Pulmonary capillary hemangiomatosis is a rare vascular proliferative disease of unknown etiology. The common clinical features are slowly progressive and finally fatal pulmonary hypertension. The clinical diagnosis is usually difficult. Because most reported cases are of autopsy, little is known about its incipient lesion and natural history. Presented herein is a case of pulmonary capillary hemangiomatosis incidentally detected in a surgically resected lung for a metastatic colon cancer. The patient was a 60-year-old Japanese woman with a history of sigmoid colon cancer 3 years previously. The patient had undergone a right lower lobectomy for a metastatic tumor in the hilar region and a thoracoscopic tumorectomy of the peripheral area of the left upper lobe. Except for an episode of hemoptysis 2 weeks prior to the lung surgery, there were no other clinical symptoms characteristic of pulmonary capillary hemangiomatosis. The non-tumor area of right lower lobe showed multiple foci of capillary proliferation affecting alveolar walls, interlobular septa and pleura associated with patchy hemorrhage. There was a minor degree of vascular and bronchial involvement by capillary proliferation. It is suggested this particular case is an incidentally detected clinically incipient stage of pulmonary capillary hemangiomatosis. Passive congestion secondary to metastatic colon cancer in the hilar region may have contributed to the pathogenesis of this lesion. PMID- 16704502 TI - Taxonomy of health networks and systems: a reassessment. AB - OBJECTIVE: To assess a widely recognized multihospital system taxonomy. DATA SOURCES: The original taxonomy was based on American Hospital Association (AHA) Annual Survey Data for the years 1994 and 1995 and a reexamined version, on 1998 AHA data. STUDY DESIGN: We assess the appropriateness of using data designed to capture local hospital/system interrelationships to develop a taxonomy of multihospital systems. DATA ABSTRACTION METHODS: The original and reexamined taxonomies used dichotomous measures of service availability, physician practice ownership, and managed care offerings. PRINCIPAL FINDINGS: The data and measures used to formulate the taxonomy are not appropriate for classifying multihospital systems at the company level. CONCLUSIONS: Taxonomic studies of multihospital systems are very much needed; future taxonomic studies should make clear distinctions between systems at local versus company levels. PMID- 16704503 TI - Rejoinder to taxonomy of health networks and systems: a reassessment. PMID- 16704505 TI - Operational failures and interruptions in hospital nursing. AB - OBJECTIVE: To describe the work environment of hospital nurses with particular focus on the performance of work systems supplying information, materials, and equipment for patient care. DATA SOURCES: Primary observation, semistructured interviews, and surveys of hospital nurses. STUDY DESIGN: We sampled a cross sectional group of six U.S. hospitals to examine the frequency of work system failures and their impact on nurse productivity. DATA COLLECTION: We collected minute-by-minute data on the activities of 11 nurses. In addition, we conducted interviews with six of these nurses using questions related to obstacles to care. Finally, we created and administered two surveys in 48 nursing units, one for nurses and one for managers, asking about the frequency of specific work system failures. PRINCIPAL FINDINGS: Nurses we observed experienced an average of 8.4 work system failures per 8-hour shift. The five most frequent types of failures, accounting for 6.4 of these obstacles, involved medications, orders, supplies, staffing, and equipment. Survey questions asking nurses how frequently they experienced these five categories of obstacles yielded similar frequencies. For an average 8-hour shift, the average task time was only 3.1 minutes, and in spite of this, nurses were interrupted mid-task an average of eight times per shift. CONCLUSIONS: Our findings suggest that nurse effectiveness can be increased by creating improvement processes triggered by the occurrence of work system failures, with the goal of reducing future occurrences. Second, given that nursing work is fragmented and unpredictable, designing processes that are robust to interruption can help prevent errors. PMID- 16704506 TI - Using public reports of patient satisfaction for hospital quality improvement. AB - OBJECTIVE: To explore the impact of statewide public reporting of hospital patient satisfaction on hospital quality improvement (QI), using Rhode Island (RI) as a case example. DATA SOURCE: Primary data collected through semi structured interviews between September 2002 and January 2003. STUDY DESIGN: The design is a retrospective study of hospital executives at all 11 general and two specialty hospitals in RI. Respondents were asked about hospital QI activities at several points throughout the public reporting process, as well as about hospital structure and processes to accomplish QI. Qualitative analysis of the interview data proceeded through an iterative process to identify themes and categories in the data. PRINCIPAL FINDINGS: Data from the standardized statewide patient satisfaction survey process were used by hospitals to identify and target new QI initiatives, evaluate performance, and monitor progress. While all hospitals fully participated in the public reporting process, they varied in the stage of development of their QI activities and adoption of the statewide standardized survey for ongoing monitoring of their QI programs. Most hospitals placed responsibility for QI within each department, with results reported to top management, who were perceived as giving strong support for QI. The external environment facilitated QI efforts. CONCLUSION: Public reporting of comparative data on patient views can enhance and reinforce QI efforts in hospitals. The participation of key stakeholders facilitated successful implementation of statewide public reporting. This experience in RI offers lessons for other states or regions as they move to public reporting of hospital quality data. PMID- 16704507 TI - The effect of cuts in medicare reimbursement on hospital mortality. AB - OBJECTIVE: To determine if patients treated at hospitals under different levels of financial strain from the Balanced Budget Act (BBA) of 1997 had differential changes in 30-day mortality, and whether vulnerable patient populations such as the uninsured were disproportionately affected. DATA SOURCE: Hospital discharge data from all general acute care hospitals in Pennsylvania from 1997 to 2001. STUDY DESIGN: A multivariate regression analysis was performed retrospectively on 30-day mortality rates, using hospital discharge data, hospital financial data, and death certificate information from Pennsylvania. DATA COLLECTION: We used 370,017 hospital episodes with one of four conditions identified by the Agency for Healthcare Research and Quality as inpatient quality indicators were extracted. PRINCIPAL FINDINGS: The average magnitude of Medicare payment reduction on overall net revenues was estimated at 1.8 percent for hospitals with low BBA impact and 3.6 percent for hospitals with a high impact in 1998, worsening to 2 and 4.8 percent, respectively, by 2001. Operating margins decreased significantly over the time period for all hospitals (p<.05). While unadjusted mortality rates demonstrated a disproportionate rise in mortality for patients from high impact hospitals from 1997 to 2000, adjusted analyses show no consistent, significant difference in the rate of change in mortality between high-impact and low-impact hospitals (p=.04-.94). Similarly, uninsured patients did not experience greater increases in mortality in high-impact hospitals relative to low-impact hospitals. CONCLUSIONS: An analysis of hospitalizations in the Commonwealth of Pennsylvania did not find an adverse impact of increased financial strain from the BBA on patient mortality either among all patients or among the uninsured. PMID- 16704509 TI - Rules for medical markets: the impact of medicare contractors on coverage policies. AB - OBJECTIVE: Examine Medicare's local contractors' claim payment rules, focusing on how technology affects the balancing of competing demands to respond to local medical markets (rule heterogeneity) with concerns about national consistency in payment rules (rule homogeneity). DATA SOURCES: Local medical review policies (LMRPs) posted in policy sets by contractor organizations on the Centers for Medicare and Medicaid Services (CMS) website and a survey of Contractor Medical Directors. STUDY DESIGN: We classified LMRPs based on type (NT=new technology; TE=technology extensions, and UM=utilization management), and examined the effect of technology type on LMRP focus, evidence use, policy revisions, implementation speed, and reference material citation characteristics of LMRPs using multivariate analysis. PRINCIPAL FINDINGS: NT policies were more homogenous, as were policies among contractors related through multistate affiliation or through informal networks. UM policies were more heterogeneous. NT policies were more likely than UM policies to cite research journals as evidence while UM policies were more likely to cite medical reference materials. CONCLUSIONS: Coverage policies associated with new technologies diffuse rapidly and are homogenous compared to utilization management coverage policies. This suggests that new technology policies are responsive to the development of new technologies at the national level. In contrast, utilization management policies are responsive to local heterogeneity in health care practice. Congress has mandated reforms to the contracting process to achieve consistency and reduce duplication. Our data elucidate the nature and sources of variation and will help policymakers strike a balance between homogeneity and local adaptation. PMID- 16704508 TI - The effect of changes in hospital reimbursement on nurse staffing decisions at safety net and nonsafety net hospitals. AB - RESEARCH OBJECTIVE: The financial savings from the Balanced Budget Act (BBA) are attractive to policy makers, but such savings come at a cost. We measure changes in nurse staffing at hospitals related to potential declines in reimbursement through the BBA. STUDY DESIGN: Following Hadley, Zuckerman, and Feder (1989), we define a fiscal pressure index (FPI) to measure the differential effect of the BBA. We estimate the effect of the FPI on the number of full-time equivalent registered nurses (RN) and licensed practical nurses (LPN) per adjusted patient day using American Hospital Association (AHA) data of a panel of hospitals from 1996 to 2001. The AHA data are combined with the Area Resource Files and health maintenance organizations penetration data. We control for hospital heterogeneity using fixed effects. POPULATION STUDIED: All urban short-term general hospitals that responded to the staffing and uncompensated care questions in the AHA survey between 1996 and 2001. We define safety net hospitals as those with a high ratio of uncompensated costs to total hospital expenses (see, e.g., Zuckerman et al. 2001). PRINCIPAL FINDINGS: We find that the nonsafety net hospitals that were most susceptible to the provisions of the BBA experienced a decline in RN staffing ratios about twice the rate of the nonsafety net hospitals that were least susceptible to the BBA. We are unable to detect an effect of the BBA on staffing at safety net hospitals. CONCLUSIONS: RN and LPN staffing levels per adjusted patient day declined, on average, between 1996 and 2001. Within the context of the general decline, we find that RN staffing per adjusted patient day declined even more at nonsafety net hospitals that were most susceptible to lower reimbursement related to the BBA. Thus, there was a small but statistically significant incremental effect of potential BBA losses on RN staffing at hospitals that were expected to be affected most. This incremental decline represented about a 6 percent increase in nurse workload that in isolation might not affect quality. Nevertheless, the BBA contributed to the contemporaneous trends toward higher nurse workloads that could have deleterious effects on quality. In contrast, safety net hospitals did not respond to the provisions of the BBA by reducing staffing ratios. This conclusion is tempered by the fact that we have few safety net hospitals in the sample. PMID- 16704510 TI - Access to health care services for the disabled elderly. AB - OBJECTIVE: To determine whether difficulty walking and the strategies persons use to compensate for this deficit influenced downstream Medicare expenditures. DATA SOURCE: Secondary data analysis of Medicare claims data (1999-2000) for age eligible Medicare beneficiaries (N=4,997) responding to the community portion of the 1999 National Long Term Care Survey (NLTCS). STUDY DESIGN: Longitudinal cohort study. Walking difficulty and compensatory strategy were measured at the 1999 NLTCS, and used to predict health care use as measured in Medicare claims data from the survey date through year-end 2000. DATA EXTRACTION: Respondents to the 1999 community NLTCS with complete information on key explanatory variables (walking difficulty and compensatory strategy) were linked with Medicare claims to define outcome variables (health care use and cost). PRINCIPAL FINDINGS: Persons who reported it was very difficult to walk had more downstream home health visits (1.1/month, p<.001), but fewer outpatient physician visits ( 0.16/month, p<.001) after controlling for overall disease burden. Those using a compensatory strategy for walking also had increased home health visits/month (0.55 for equipment, 1.0 for personal assistance, p<.001 for both) but did not have significantly reduced outpatient visits. Persons reporting difficulty walking had increased downstream Medicare costs ranging from 163 US dollars to 222 US dollars/month (p<.001) depending upon how difficult walking was. Less than half of the persons who used equipment to adapt to walking difficulty had their difficulty fully compensated by the use of equipment. Persons using equipment that fully compensated their difficulty used around 300 US dollars/month less in Medicare-financed costs compared with those with residual difficulty. CONCLUSIONS: Difficulty walking and use of compensatory strategies are correlated with the use of Medicare-financed services. The potential impact on the Medicare program is large, given how common such limitations are among the elderly. PMID- 16704511 TI - The effect of private insurance on the health of older, working age adults: evidence from the health and retirement study. AB - OBJECTIVE: Primarily, to determine if the presence of private insurance leads to improved health status, as measured by a survey-based health score. Secondarily, to explore sensitivity of estimates to adjustments for endogeneity. The study focuses on adults in late middle age who are nearing entry into Medicare. DATA SOURCES: The analysis file is drawn from the Health and Retirement Study, a national survey of relatively older adults in the labor force. The dependent variable, an index of 5 health outcome items, was obtained from the 1996 survey. Independent variables were obtained from the 1992 survey. State-level instrumental variables were obtained from the Area Resources File and the TAXSIM file. The final sample consists of 9,034 individuals of which 1,540 were uninsured. STUDY DESIGN: Estimation addresses endogeneity of the insurance participation decision in health score regressions. In addition to ordinary least squares (OLS), two models are tested: an instrumental variables (IV) model, and a model with endogenous treatment effects due to Heckman (1978). Insurance participation and health behaviors enter with a lag to allow their effects to dissipate over time. Separate regressions were run for groupings of chronic conditions. PRINCIPAL FINDINGS: The OLS model results in statistically significant albeit small effects of insurance on the computed health score, but the results may be downward biased. Adjusting for endogeneity using state-level instrumental variables yields up to a six-fold increase in the insurance effect. Results are consistent across IV and treatment effects models, and for major groupings of medical conditions. The insurance effect appears to be in the range of about 2-11 percent. There appear to be no significant differences in the insurance effect for subgroups with and without major chronic conditions. CONCLUSIONS: Extending insurance coverage to working age adults may result in improved health. By conjecture, policies aimed at expanding coverage to this population may lead to improved health at retirement and entry to Medicare, potentially leading to savings. However, further research is needed to determine whether similar results are found when alternative measures of overall health or health scores are used. Future research should also explore the use of alternative instrumental variables. Preliminary results provide no justification for targeting certain subgroups with susceptibility to certain chronic conditions rather than broad policy interventions. PMID- 16704512 TI - Community-level uninsurance and the unmet medical needs of insured and uninsured adults. AB - OBJECTIVE: To examine the relationship between community-level uninsurance rates and the self-reported unmet medical needs of insured and uninsured adults in the U.S. DATA SOURCES: 2000-2001 Community Tracking Study, which includes data from 60 randomly selected U.S. communities. The sample is representative of the contiguous U.S. states. STUDY DESIGN: Multilevel logistic regressions were employed to investigate whether the local uninsurance rate was related to having reported unmet medical needs within the last year. The models also included individual and community variables that could be potentially related to both community uninsurance rates and having reported unmet medical needs. PRINCIPAL FINDINGS: The community uninsurance rate was positively associated with having reported unmet medical needs, but only for insured adults. On average, a five percentage point increment in the local uninsured population is associated with a 10.5 percent increase in the likelihood that an insured adult will report having unmet medical needs during the 12-month period studied. CONCLUSION: Local health care delivery systems seem to be negatively affected by high uninsurance rates. These effects could have negative consequences for health care access, even for individuals who are themselves insured. PMID- 16704513 TI - When to combine hypotheses and adjust for multiple tests. AB - OBJECTIVE: To provide guidelines for identifying composite hypotheses and addressing the probability of false rejection for multiple hypotheses. DATA SOURCES AND STUDY SETTING: Examples from the literature in health services research are used to motivate the discussion of composite hypothesis tests and multiple hypotheses. METHODS: This article is a didactic presentation. PRINCIPAL FINDINGS: It is not rare to find mistaken inferences in health services research because of inattention to appropriate hypothesis generation and multiple hypotheses testing. Guidelines are presented to help researchers identify composite hypotheses and set significance levels to account for multiple tests. CONCLUSIONS: It is important for the quality of scholarship that inferences are valid: properly identifying composite hypotheses and accounting for multiple tests provides some assurance in this regard. PMID- 16704514 TI - Who searches the internet for health information? AB - OBJECTIVE: To determine what types of consumers use the Internet as a source of health information. DATA SOURCES: A survey of consumer use of the Internet for health information conducted during December 2001 and January 2002. STUDY DESIGN: We estimated multivariate regression models to test hypotheses regarding the characteristics of consumers that affect information seeking behavior. DATA COLLECTION: Respondents were randomly sampled from an Internet-enabled panel of over 60,000 households. Our survey was sent to 12,878 panel members, and 69.4 percent of surveyed panel members responded. We collected information about respondents' use of the Internet to search for health information and to communicate about health care with others using the Internet or e-mail within the last year. PRINCIPAL FINDINGS: Individuals with reported chronic conditions were more likely than those without to search for health information on the Internet. The uninsured, particularly those with a reported chronic condition, were more likely than the privately insured to search. Individuals with longer travel times for their usual source of care were more likely to use the Internet for health related communication than those with shorter travel times. CONCLUSIONS: Populations with serious health needs and those facing significant barriers in accessing health care in traditional settings turn to the Internet for health information. PMID- 16704515 TI - Geographic variation in the prescription of schedule II opioid analgesics among outpatients in the United States. AB - OBJECTIVE: To measure geographic variation in opioid use in a large, commercially insured, outpatient population in the United States. DATA SOURCES: Outpatient prescription drug claims database of a national pharmaceutical benefit manager for 7,873,337 subjects with at least one prescription drug claim in 2000. STUDY DESIGN: We measured the period prevalence of claims for opioid analgesics and controlled-release oxycodone at the state level. We measured geographic variation using the weighted coefficient of variation and systematic component of variation. In county-level multivariable regression, we explored associations between potential explanatory variables and claims for opioid analgesics and controlled-release oxycodone. PRINCIPAL FINDINGS: A total of 567,778 (64.2 per 1,000 total claims) were for oral opioid analgesics. Claim rates by state ranged from <20 to >100 claims per 1,000 total claims. States with long-standing prescription monitoring programs had among the lowest rates. In the county-level data, presence of a statewide prescription monitoring program and proportions of the population aged 15-24 and 65 years and older were independently and negatively associated with claim rates for all opioid analgesics. Surgeons per 1,000, proportion of the population reporting illicit drug use, and proportion who were female were independently and positively associated with claim rates for all opioid analgesics. Only the proportion of the population aged 25-34 and number of surgeons per 1,000 were independently and positively associated with claim rates for oxycodone. CONCLUSIONS: Claim rates for opioid analgesics vary significantly by state. Presence of a statewide prescription monitoring program is associated with lower claim rates at the county level. Future research should use individual-level data to assess whether these findings reflect a reduction in abuse and diversion or suboptimal treatment of pain. PMID- 16704517 TI - Factors influencing female registered nurses' work behavior. AB - OBJECTIVE: To analyze factors that are related to whether registered nurses (RNs) work (WK) or do not work (NW) in nursing; and if the RN works, whether she works full- (FT) or part-time (PT). DATA SOURCES: Secondary data from National Sample Survey of Registered Nurses 2000 (NSSRN), the InterStudy Competitive Edge Part III Regional Market Analysis (2001), and the Area Resource File (2002). STUDY DESIGN: Using a cross-sectional design we tested the relationship between WK or NW and FT or PT; and demographic, job-related, and metropolitan statistical area (MSA)-level variables. DATA COLLECTION/EXTRACTION METHODS: We combined the data sources noted above to produce the analytic sample of 25,471 female RNs. PRINCIPAL FINDINGS: Working in nursing is not independent of working FT or PT. Age (55 and older), other family income, and prior other work experience in health care are negatively related to working as an RN. The wage is not related to working as an RN, but negatively influences FT work. Age, children, minority status, student status, employment status, other income, and some job settings have a negative impact on working FT. Previous health care work has a positive effect on whether married RNs worked. Married RNs who are more dissatisfied are less likely to work FT. A greater number of market-level factors influence FT/PT than WK/NW behavior. CONCLUSIONS: An important contribution of this study is demonstrating that MSA-level variables influence RN work behavior. The market environment seems to have little effect on whether a nurse works, but is influential on how much the nurse works, and has differential effects on married versus single nurses. PMID- 16704518 TI - Health policy roundtable: how are Medicaid and SCHIP weathering the storm? PMID- 16704519 TI - Health policy roundtable: producing and adapting research syntheses for use by health-system managers and public policymakers. PMID- 16704520 TI - Toward a more reliable federal survey for tracking health insurance coverage and access. AB - OBJECTIVE: Examination of the extent to which federal surveys provide the data needed to estimate the coverage/cost impacts of policy alternatives to address the problem of uninsurance. DATA SOURCES/STUDY SETTING: Assessment of the major federal household surveys that regularly provide information on health insurance and access to care based on an examination of each survey instrument and related survey documentation and the methodological literature. STUDY DESIGN: Identification of the data needed to address key policy questions on insurance coverage, assessment of how well existing surveys meet this need, definition of the critical elements of an ideal survey, and examination of the potential for building on existing surveys. DATA COLLECTION/EXTRACTION METHODS: Collection and critical assessment of pertinent survey documentation and methodological studies. PRINCIPAL FINDINGS: While all the federal surveys examined provide valuable information, the information available to guide key policy decisions still has major gaps. Issues include measurement of insurance coverage and critical content gaps, inadequate sample sizes to support precise state and substate estimates, considerable delays between data collection and availability, and concerns about response rates and item nonresponse. Our assessment is that the Current Population Survey (CPS) and the National Health Interview Survey could be most readily modified to address these issues. CONCLUSIONS: The vast resources devoted to health care and the magnitude of the uninsurance problem make it critical that we have a reliable source for tracking health care and coverage at the national and state levels and for major local areas. It is plausible that this could be more cost effectively done by building on existing surveys than by designing and fielding a new one, but further research is needed to make a definitive judgment. At a minimum, the health insurance information collected on the CPS should be revised to address existing measurement problems. PMID- 16704521 TI - Meeting the need for state-level estimates of health insurance coverage: use of State and Federal survey data. AB - OBJECTIVE: Critically review estimates of health insurance coverage available from different sources, including the federal government, state survey initiatives, and foundation-sponsored surveys for use in state policy research. STUDY SETTING AND DESIGN: We review the surveys in an attempt to flesh out the current weaknesses of survey data for state policy uses. The main data sources assessed in this analysis are federal government surveys (such as the Current Population Survey's Annual Social and Economic Supplement, and the National Health Interview Survey), foundation-supported surveys (National Survey of America's Families, and the Community Tracking Survey), and state-sponsored surveys. PRINCIPAL FINDINGS: Despite information on estimates of health insurance coverage from six federal surveys, states find the data lacking for state policy purposes. We document the need for state representative data on the uninsured and the recent history of state data collection efforts spurred in part by the Health Resources Services Administration State Planning Grant program. We assess the state estimates of uninsurance from the Current Population Survey and make recommendations for a new consolidated federal survey with better state representative data. CONCLUSIONS: We think there are several options to consider for coordinating a federal and state data collection strategy to inform state and national policy on coverage and access. PMID- 16704522 TI - Commentary--how to improve Federal health data for coverage, access, and State specific needs. PMID- 16704523 TI - Commentary--Improving Federal health data: the essential partnership between researcher and policy maker. PMID- 16704524 TI - Immunoglobulin class M anti-hepatitis B core antigen for serodiagnosis of acute hepatitis: pitfalls and recommendations. PMID- 16704525 TI - Histopathological diagnosis of microscopic colitis. AB - A typical symptom of microscopic colitis (MC) is chronic watery diarrhea with normal endoscopic findings and characteristic inflammatory changes in histopathology. Treatment of the disease is mainly empiric. MC has two main subtypes: lymphocytic colitis and collagenous colitis. There are also untypical histopathological forms of MC: MC with giant cells, MC not otherwise specified (NOS) and cryptal lymphocytic coloproctitis. Some other histopathological changes in MC have been observed, especially Paneth cell hyperplasia or epithelial degeneration. Eosinophilic colitis, acute colitis, amyloidosis, ulcerative colitis and Crohn's disease should be taken into consideration in differential diagnosis. The most reliable biopsy material for histopathological examination are samples obtained from transverse colon. Some studies proved that treatment of MC makes it possible to reduce not only clinical, but also histopathological, manifestations. PMID- 16704526 TI - Comparative analysis between psychological and endoscopic profiles in patients with gastroesophageal reflux disease: a prospective study based on screening endoscopy. AB - BACKGROUND: Associations between psychological and endoscopic profiles are not clearly validated among the heterogeneous patients with gastroesophageal reflux disease (GERD). The purpose of the present paper was therefore to identify any associations by means of cross-sectional study. METHODS: Consecutive participants in a health screening program were enrolled. Definition and severity of erosive esophagitis were assessed with Los Angeles classification. Frequency and severity of psychological symptoms were measured with a 30-item Brief Symptom Rating Scale (BSRS) and personality traits with a short form of the Maudsley Personality Inventory (MPI). Statistic analyses were performed based on the presence of GERD symptoms or endoscopic esophagitis. RESULTS: A total of 4600 participants were recruited. There were 1331 subjects (29%) with manifestations suggesting GERD, including non-erosive reflux disease (NERD) in 488 (10.6%), symptomatic erosive esophagitis (SEE) in 164 (3.6%), and asymptomatic erosive esophagitis (AEE) in 679 (14.8%). The BSRS parameters were significantly higher in symptomatic subjects (i.e. NERD and SEE subjects; P < 0.001); neuroticism scores were also higher (P < 0.001), but extroversion scores (P < 0.001) were lower than those of asymptomatic subjects. Following logistic regression analysis, independent risk factors for GERD symptoms were female gender (odds ratio [OR]: 1.596; 95% confidence interval [CI]: 1.303-1.955) and higher neuroticism scores (OR: 1.046; 95%CI: 1.032-1.06). For erosive esophagitis, independent risk factors were male gender (OR: 2.943; 95%CI: 2.359-3.671) and higher body mass index scores (OR: 1.098; 95%CI: 1.069-1.127). CONCLUSIONS: Psychological characteristics predict likelihood of GERD symptoms but not structural state of esophagus. Male gender and obesity are risk factors for erosive esophagitis; whereas female gender and neuroticism are more likely to be associated with GERD symptoms. PMID- 16704527 TI - Effect of chronic dietary ethanol in the promotion of N-nitrosomethylbenzylamine induced esophageal carcinogenesis in rats. AB - BACKGROUND: The pathogenetic correlation between chronic alcohol consumption and development of esophageal cancer is not clear. The role of alcohol abuse in the carcinogenic action of N-nitrosomethylbenzylamine, which induces tumors in the esophagus, has been evaluated. METHODS: Twenty male rats were fed liquid diets containing ethanol or carbohydrates for 30 weeks. N-nitrosomethylbenzylamine (0.1 mg/kg, twice a week) was injected i.p. from the 9th to 19th week. The pair feeding was stopped at 9.00 am and N-nitrosomethylbenzylamine was administered at 10.00 am. Ethanol was not detected in the blood at the time of injection. Liquid diets were provided again at 3 pm until 9 am next day. The animals were killed at the end of the 30th week. The esophagi were collected and examined for visible tumors. The tissue sections were stained for histology and CYP2E1expression. RESULTS: While 5-8 esophageal squamous polyps developed in all rats in the ethanol group, only one polyp each was formed in five out of the 10 rats in the control group. The size of the polyps was significantly larger in the ethanol group, when compared to the control group. Invasive squamous cell carcinoma was also observed in 50% of the animals in the ethanol group. Cytochrome P4502E1 (CYP2E1) staining demonstrated marked expression in the esophageal mucosa in the ethanol group, but not in the control group. CONCLUSIONS: The increased expression of CYP2E1 induced by chronic ethanol consumption promotes the development of N-nitrosomethylbenzylamine-induced esophageal tumorigenesis. However, the molecular mechanism of the increased production of esophageal tumors during alternative administration of N-nitrosomethylbenzylamine and ethanol is not clear. PMID- 16704528 TI - Impact of loss of heterozygosity of encoding phosphate and tensin homolog on the prognosis of gastric cancer. AB - BACKGROUND AND AIM: Encoding phosphate and tensin homolog (PTEN) is a cancer suppressor gene and it has been assumed that gene mutation and loss of heterozygosity (LOH) occurs frequently in various types of carcinoma. However, the role of LOH of PTEN and its outcome variables in gastric cancer have not been well established. In the present study, we investigated the roles of PTEN, LOH and their outcomes. METHODS: Fresh frozen tumor samples from 119 gastric cancer patients with a primary diagnosis of gastric carcinoma were evaluated for LOH of PTEN using an automated sequencer. Results were compared with pathological parameters. The median follow-up period was 559 days. RESULTS: Loss of heterozygosity of PTEN was observed in 17.1% of patients (13/76) diagnosed with gastric cancer. No particular relationship was found with any clinicopathological factor. However, the prognosis of patients with LOH of PTEN was significantly poor. Multivariate analyses revealed that vascular invasion, invasion depth, LOH of PTEN, histology and lymph node metastasis were correlated with survival of the patient. CONCLUSIONS: Even though mutation of PTEN in gastric cancer has rarely been reported, according to our findings, LOH of PTEN frequently occurs in gastric cancers and is correlated with disease-related deaths. The LOH of PTEN is an independent prognostic factor and PTEN is a candidate as a haploinsufficient tumor suppressor in gastric cancers. PMID- 16704529 TI - Use of polymerase chain reaction in the diagnosis of abdominal tuberculosis. AB - BACKGROUND: Early diagnosis and prompt treatment of abdominal tuberculosis is vitally important as it greatly reduces disease and treatment related morbidity and even mortality in extreme cases. A polymerase chain reaction (PCR) test was evaluated for its feasibility as a diagnostic tool in abdominal tuberculosis (TB) in the Indian scenario. METHODS: PCR for the identification of M. tuberculosis amplified a 340 bp nucleotide sequence located within the 38 kDa protein gene of M. tuberculosis. Tissues for processing were obtained from patients suspected to have abdominal TB. These were from various sources such as abdominal lymph nodes, segments of intestine and bowel obtained at various times and in different ways such as laparoscopy, colectomy, bowel and lymph node resection. Fifty such patients had their tissues sent for PCR. RESULTS: PCR results were compared with histopathology (HP). Of the 50 samples, 31 were positive for abdominal TB by HP whereas 30 were positive by PCR. Twenty-four of these were positive for both HP and PCR while of the seven samples positive for HP, five were negative and two gave inhibition by PCR. Six samples negative by HP were positive by PCR. CONCLUSION: This study demonstrates that PCR can be used as an effective tool to diagnose abdominal TB. PMID- 16704530 TI - Effects of Helicobacter pylori eradication therapy on gastric emptying measured using the 13C-octanoic acid breath test and the acetaminophen method. AB - BACKGROUND: The relationship between Helicobacter pylori eradication and gastric emptying has been reported; however, the effect of eradication therapy on gastric emptying is still unclear. This study evaluated the relationship between three gastric emptying techniques, the scintigraphic technique, the 13C-octanoic acid breath test, and the acetaminophen method, measured simultaneously, and the effect of H. pylori eradication therapy on gastric emptying and abdominal symptoms in patients with functional dyspepsia who were H. pylori positive. METHODS: Fifty-three consecutive patients with positive H. pylori infection were enrolled in this study. In the first 14 patients, gastric emptying was measured using the three gastric emptying techniques. In 42 patients cured of H. pylori infection, the 13C-octanoic acid breath test and the acetaminophen method were performed before and 3 months after eradication. RESULTS: Significant correlations were found between the scintigraphic technique, the 13C-octanoic acid breath test, and the acetaminophen method. Gastric emptying determined by the 13C-octanoic acid breath test and the acetaminophen method was not changed after eradication, on average. In 14 (33.3%) patients a decrease in symptom score after eradication was observed. In four (9.5%) patients, accelerated gastric emptying after eradication may have led to a reduction in the abdominal symptoms. CONCLUSIONS: The 13C-octanoic acid breath test and the acetaminophen method are appropriate for investigating gastric emptying. A causal relationship between improvement of symptoms and accelerated gastric emptying was not found, and the efficacy of H. pylori eradication therapy in patients with functional dyspepsia was minimally exhibited. PMID- 16704531 TI - Interaction of carbohydrate metabolism and rat liquid gastric emptying in sustained running. AB - BACKGROUND: Moderate to severe running usually leads to gastrointestinal dysmotility and critical energy exhaustion. It is unknown whether the carbohydrate metabolism of runners can influence gastric emptying (GE). Using a running rat model, the present study explored the impact of exercise/carbohydrate metabolism on liquid GE. METHODS: Rats were put on the runways of a moving treadmill for 1 h. Trained rats underwent daily running 5 days a week for 4 weeks. Untrained rats were those put on a quiet treadmill to serve as sham exercise. On the motility study day, trained and untrained rats ran for 45 min. After orogastric feeding of radiochromium marker, they resumed running for an additional 15 min and were then killed in order to measure GE. Another group of trained and untrained rats received lactate infusion for 1 h in the quiet condition to measure their GE. The third group of rats received glucose infusion during running to measure GE. RESULTS: Running of untrained (P < 0.05) and trained (P < 0.01) rats enhanced GE compared to sham exercise. Running for the untrained rats rather than the trained counterparts had diminished plasma glucose level (P < 0.05). Running also elevated plasma lactate levels for both untrained (P < 0.001) and trained rats (P < 0.01). Lactate infusion delayed GE in untrained (P < 0.01) and trained rats (P < 0.05). Glucose infusion of untrained rats during running was not only to correct hypoglycemia (P < 0.01) but also to restore their enhanced GE (P < 0.01). CONCLUSIONS: Running-induced hypoglycemia, rather than lactate accumulation, is one of the essential factors leading to enhanced liquid GE in untrained rats. PMID- 16704532 TI - Surface structure of antral gastric mucosa represents the status of histologic gastritis: fundamental evidence for the evaluation of antral gastritis by magnifying endoscopy. AB - BACKGROUND: Recent studies have demonstrated the diagnostic potential of magnifying endoscopy in cases of histologic gastritis. The aim of the present study was to clarify whether the mucosal surface structure reflects the degree of histologic gastritis below the surface. METHODS: Gastric biopsy specimens were obtained from 1018 Japanese patients and were stained with hematoxylin and eosin. In 863 sections examined, gastritis was graded by means of the updated Sydney system, and the surface structure was classified as one of four types: flat, irregular, papillary, or non-structured. In addition, 103 patients underwent gastric examination by magnifying gastroscopy. RESULTS: The surface structure of most biopsy sections with normal mucosa and no Helicobacter pylori infection was classified as the flat type. Grades of histological gastritis were statistically lower in flat-type sections than in other types. Histologic gastritis was found in 91% of sections with a non-flat surface structure. Helicobacter pylori infection was confirmed in 96% of these cases. Most biopsy sections in patients with abnormal magnifying endoscopy features were of the non-flat type and showed histologic gastritis. CONCLUSIONS: The surface structure of the gastric mucosa reflects the status of histologic gastritis. Magnifying gastroscopy could be a useful non-invasive method of diagnosing histologic gastritis. PMID- 16704533 TI - Intussusception: trends in clinical presentation and management. AB - BACKGROUND: The association of a rotavirus vaccine and intussusception has renewed interest in understanding the incidence, clinical presentation and outcome of intussusception. METHODS: A retrospective chart review of all patients diagnosed with intussusception at Royal Children's Hospital, Melbourne over a 6.5 year period (1 January 1995-30 June 2001) was conducted using patients identified by a medical record database (ICD-9-CM code 560.0 1993-1997; ICD-10-CM code 56.1 1998-2001). Patient profile, clinical presentation, diagnosis methods, treatment and outcome were analyzed and compared to data previously reported on children with intussusception at the same hospital during 1962-1968. RESULTS: The hospitalization rate for primary idiopathic intussusception increased marginally from 0.19 to 0.27 per 1000 live births during the period 1962-1968 to 1995-2001. Most patients (80%) were <12 months of age (median age 7 months, range 2-72 months). The combination of abdominal pain, lethargy and vomiting was reported in 78% of infants. Air enema confirmed the diagnosis of intussusception in 186 of 191 cases (97%) and air reduction was successful in most cases (82%). Factors associated with increased risk of intestinal resection included abdominal distension (32%), bowel obstruction on abdominal X-ray (27%) and hypovolemic shock (40%). No mortality was observed in the present study. CONCLUSIONS: Over the past 40 years at Royal Children's Hospital, Melbourne the hospitalization rate due to primary idiopathic intussusception has marginally increased from 0.19 to 0.27 per 1000 live births. Diagnosis and treatment using air enema has been highly successful, resulting in a reduction in patients requiring surgery and reduced hospital stays. PMID- 16704534 TI - Association of low density lipoprotein receptor related protein-associated protein (LRPAP1) gene insertion/deletion polymorphism with gallstone disease. AB - BACKGROUND AND AIM: Gallstones are byproducts of cholesterol supersaturated bile. Various studies have indicated that there might be a genetic predisposition to the disease. Receptor-associated protein (RAP) is a molecular chaperone for low density lipoprotein receptor-related protein (LRP), which plays a key role in cholesterol metabolism. Intron 5 insertion/deletion polymorphism of RAP gene (LRPAP1) has been implicated in other diseases sharing etiology with gallstone disease (GSD). METHODS: To analyze the association of insertion/deletion polymorphism in GSD, 130 gallstone patients and 202 healthy subjects took part in the present study. For genotyping, polymerase chain reaction was followed by 2% agarose gel electrophoresis. RESULTS: The results showed that frequencies of D and I allele were 65.77% and 34.23% in patients, 76.24% and 23.76% in controls, respectively. Frequency of I allele was significantly higher in the patient group than in the control group (P = 0.003). CONCLUSION: In the present study I (insertion) allele was found to be associated with GSD. PMID- 16704535 TI - Expression of nerve growth factor receptors is correlated with progression and prognosis of human pancreatic cancer. AB - BACKGROUND AND AIM: The aim of the present study was to investigate the prognostic value of the two types of nerve growth factor receptors (NGFR), namely high-affinity receptor TrkA and low-affinity receptor p75NGFR, in pancreatic cancer. METHODS: The mRNA expression of NGFR for TrkA and p75NGFR was examined in 56 human primary pancreatic cancers using real-time quantitative reverse transcription-polymerase chain reaction. RESULTS: Nerve growth factor (NGF) receptors were found in all tumor specimens. It appears that the growth of pancreatic cancer cells stimulated by NGF depended on the expression levels and the ratio of TrkA to p75NGFR. TrkA and p75NGFR were negatively correlated and both were associated with abdominal or back pain and perineural invasion. Regarding this, patients with high TrkA expression levels exhibited more frequent perineural invasion and a higher degree of pain, whereas the results of p75NGFR were opposite. For Cox univariate analyses in the overall survival study, high expression of p75NGFR was associated with longer overall survival, but TrkA exhibited opposite effects and included an effect on perineural invasion and pain. Histoprognostic grading, tumor size and node involvement were not prognostic factors. In Cox multivariate analyses, TrkA and p75NGFR were both prognostic parameters. CONCLUSIONS: The present study found that the expression of TrkA in pancreatic cancer is a marker of tumor aggressiveness. Conversely, we also found that elevated p75NGFR expression is associated with a favorable prognosis. We demonstrated that NGF exerts both stimulatory and inhibitory effects on pancreatic cancers, with the overall effect determined by the expression levels and the ratio of TrkA to p75NGFR. PMID- 16704536 TI - Higher cut-off index value of immunoglobulin M antibody to hepatitis B core antigen in Taiwanese patients with hepatitis B. AB - BACKGROUND: The cut-off index value of immunoglobulin M (IgM) antibody to hepatitis B core antigen (anti-HBc; AxSYM CORE-M, Abbott) for diagnosing acute hepatitis B is 1.2. A high false-positive rate of IgM anti-HBc was observed in acute flare-ups of chronic hepatitis B in Taiwanese patients. Thus the purpose of the present paper was to study the optimal index value of IgM anti-HBc in Taiwanese subjects. METHODS: The peak index values of 42 IgM anti-HBc-positive patients were collected. There were 20 acute hepatitis B patients and 22 patients with chronic hepatitis B with acute flare. The biochemical, virological, and serological data were obtained. RESULTS: There were significant differences in mean age (36 vs 47 years, P = 0.01), serum alanine aminotransferase level (2042 U/L vs 1193 U/L, P = 0.02) and peak index value of IgM anti-HBc (2.9 vs 1.5, P < 0.01) between patients with acute hepatitis B and those with acute flare of chronic hepatitis B. Eleven (50%) of 22 patients with chronic hepatitis B with acute flare had index value of >1.2. The optimal cut-off index value to differentiate acute hepatitis B from chronic hepatitis B with acute flare was 2.4 2.5, with a sensitivity of 90% and specificity of 90%. CONCLUSIONS: The cut-off index value of IgM anti-HBc to differentiate acute hepatitis B from chronic hepatitis B with acute flare among Taiwanese patients should be set at 2.4-2.5 instead of 0.8-1.2. PMID- 16704537 TI - Intrafamilial prevalence of hepatitis B virus in Western Brazilian Amazon region: epidemiologic and biomolecular study. AB - BACKGROUND: Hepatitis B is endemic in the Amazon region. METHODS: Serological markers for hepatitis B virus (HBV) were determined in 266 household members for hepatitis B surface antigen (HBsAg)-positive women (G1) and 395 household members for HBsAg-negative women (G2), randomly selected in Acre State Women's Medical Care Program, in order to evaluate the prevalence of HBV in this population. Before blood sample collection an epidemiological questionnaire was applied. RESULTS: The overall prevalence of HBV carriers (HBsAg) and exposed individuals (anti-HBc, IgG) was, respectively, 21.1% and 60.5% in G1 and 2.8% and 27.4% in G2 (P < 0.0000001). The frequency of HBsAg was higher among siblings from group G1 (75%) compared to the absence of any HBsAg-positive sibling in G2 (P < 0.00006). The HBV markers in other family members was as follows: G1 parents, 27.3% vs 4.5% (P < 0.03), sexual partners, 21.1% vs 2.5% (P < 0.04), and offspring, 10.4% vs 1.5% (P < 0.04). A low prevalence of HBsAg and anti-HBc (IgG) was observed for the last offspring of G2 mothers compared to the high prevalence among children of G1 mothers (0% vs 18.2%, P < 0.01 and 2.3% vs 59.1%, P < 0.0000005, respectively), with children younger than 1 year being the most affected. The frequency of the habit of sharing toothbrushes and the presence of at least one HBsAg carrier were higher in G1 than in G2 (P < 0.0001 and P < 0.000002), respectively. Genotypes A, D and G were found to be predominant by Innolipa test. There were cases that reacted to more than one genotype. CONCLUSION: Intrafamilial transmission of HBV is evident in the present study and is possibly associated with the presence of more than one HBV carrier in the family and the shared use of toothbrushes among household contacts. Genotype analysis confirms intrafamilial transmission. PMID- 16704538 TI - Clinicopathologic features of patients with intrahepatic cholangiocarcinoma who are seropositive for alpha-fetoprotein-L3 and those with combined hepatocellular and cholangiocarcinoma. AB - BACKGROUND: Some patients who are seropositive for lectin-reactive alpha fetoprotein (AFP-L3) have intrahepatic cholangiocarcinoma (ICC). There have been no studies regarding the features of ICC patients seropositive for AFP-L3. Thus, the purpose of the present paper was to compare the features of ICC patients from the viewpoint of two different tumor markers, AFP-L3 and carbohydrate antigen (CA) 19-9. METHODS: The ICC patients who underwent hepatectomy (n = 51) were divided into three groups, and their clinicopathologic features were compared: (i) group A, seropositive for AFP-L3 >or= 15%; (ii) group B, seropositive for CA 19-9 >or= 37 U/mL; and (iii) group C, seronegative for both AFP-L3 and CA 19-9. The features of combined hepatocellular and cholangiocarcinoma (n = 11) were also studied. RESULTS: Group A had a higher positivity rate for hepatitis viruses than group B (60%vs 20%, P < 0.05). More patients in group A were misdiagnosed as having hepatocellular carcinoma (HCC) at surgery (70%vs 5.7%, P < 0.001) who also had chronic liver disease (80%vs 25.7%, P < 0.01) than in group B. Seven, 10 and 11 of the 11 patients with combined hepatocellular and cholangiocarcinoma were seropositive for AFP-L3, CA 19-9 and hepatitis viruses, respectively. Ten were diagnosed as having HCC at surgery and nine had chronic liver disease. CONCLUSIONS: Patients with ICC seropositive for AFP-L3 and those with combined hepatocellular and cholangiocarcinoma have features close to HCC. The present study has, for the first time, identified a subgroup of ICC patients, seropositive for AFP-L3, having features close to HCC that are very different from those of the classical ICC patients seropositive for CA 19-9. PMID- 16704539 TI - Phase I study of biweekly gemcitabine followed by oxaliplatin and simplified 48-h infusion of fluorouracil/leucovorin for advanced pancreatic cancer. AB - OBJECTIVES: To evaluate the feasibility and maximal tolerated dose (MTD) of oxaliplatin of a triplet regimen consisting of gemcitabine, oxaliplatin and infusional fluorouracil (5-FU)/leucovorin (LV) (GOFL) for advanced pancreatic cancer. PATIENTS AND METHODS: Patients with histologically proven metastatic or unresectable, locally advanced pancreatic adenocarcinoma were eligible to take part in the study. The treatment consisted of fixed-rate infusion (10 mg/m2/minute) of 800 mg/m2 gemcitabine followed by 2-h infusion of oxaliplatin and then 48-h infusion of 5-FU/LV day 1 and day 15 every 4 weeks. The oxaliplatin would be evaluated at three dose levels, 65, 75 and 85 mg/m2. RESULTS: A total of 15 patients were enrolled at three dose levels. Dose-limiting toxicity of neutropenic fever and grade 4 thrombocytopenia occurred in one of each six patients at oxaliplatin dose level of 65 mg/m2 and 85 mg/m2, respectively. The MTD of oxaliplatin for this combination was 85 mg/m2. After a median four cycles of treatment, grade 3/4 neutropenia occurred in 46.7% of patients and thrombocytopenia in 13.3%. Non-hematological toxicities were generally of grade 1/2. Objective tumor response was observed in five patients (33.3%, 95% confidence interval, 6.3-60.4%). CONCLUSION: Biweekly GOFL is a feasible regimen for advanced pancreatic cancer. For further phase II studies, the recommended dose of oxaliplatin is 85 mg/m2. PMID- 16704540 TI - Effectiveness of chemical biliary duct embolization for chemical hepatectomy. AB - BACKGROUND AND AIMS: The high recurrence of hepatolithiasis and high operative trauma of hepatectomy necessitate new therapeutic approaches. Thus, this study was designed to (i) investigate the effectiveness of chemical biliary duct embolization (CBDE) for chemical hepatectomy; and (ii) to determine the mechanism of CBDE. METHODS: The median biliary ducts in rats were injected with phenol or absolute ethanol alone, or in conjunction with cyanoacrylate. The effectiveness of CBDE for chemical hepatectomy was assessed by investigating histology, in situ hybridization for Fas and transforming growth factor (TGF)-beta1, immunohistochemistry for alpha-smooth muscle actin, and reverse transcription polymerase chain reaction (RT-PCR) for procollagen I mRNA. RESULTS: Histologically, phenol or absolute ethanol plus cyanoacrylate could embolize the targeted bile duct and promote hepatic fibrosis and atrophy in the embolized lobe better than using phenol or ethanol alone. In addition, CBDE accelerated hepatocellular apoptosis via up-regulation of Fas, thereby resulting in the death of hepatocytes, which were replaced by proliferative bile ductules and collagen. Importantly, the hepatocytes disappeared completely in the periphery of the embolized lobe, thus achieving the desired effects of chemical hepatectomy. Further investigation indicated that CBDE initiated progressive fibrogenic processes of chemical hepatectomy via up-regulation of TGF-beta1, which greatly enhanced the synthesis of collagen. Indeed, higher levels of TGF-beta1 and procollagen I mRNA were observed in the phenol embolization group than in any other group. CONCLUSION: Chemical biliary duct embolization, especially using phenol plus cyanoacrylate, might achieve the effects of chemical hepatectomy. PMID- 16704541 TI - Applicability of the IAIHG scoring system to the diagnosis of antimitochondrial/anti-M2 seropositive variant form of autoimmune hepatitis. AB - BACKGROUND AND AIMS: According to the International Autoimmune Hepatitis Group (IAIHG) criteria, circulating antimitochondrial antibodies (AMA) do not support the diagnosis of autoimmune hepatitis (AIH). The aims of this study were to characterize a subset of patients with AIH who have AMA and antiM2 seropositivity, and to assess the applicability of the revised scoring system of the IAIHG in the diagnosis of this variant form of AIH. METHODS: Eighteen patients with AMA-AIH were enrolled and compared with 206 classical AIH and 85 primary biliary cirrhosis (PBC) controls. Human leukocyte antigen (HLA) class II alleles were determined by polymerase chain reaction (PCR) amplification with sequence-specific primers, and biopsies were blindly reevaluated. RESULTS: The patients with AMA-AIH were, on average, older than patients with classical AIH and had an hepatocellular pattern of elevated liver enzymes, hypergammaglobulinemia and lower levels of cholesterol, when compared with PBC controls. There were no histological signs of PBC or overlapping forms in any AMA AIH biopsies. The majority of patients with AMA-AIH carried HLA antigens associated with classical AIH (DRB1*03, n = 5; DRB1*04, n = 7, and DRB1*13, n = 6). Pretreatment scores classified all AMA-AIH patients with probable (n = 17) or improbable (n = 1) AIH. After treatment, only 28% of AMA-AIH patients reached scores for definite diagnosis, compared with 90.1% of AIH-1 and 96.4 AIH-2. In the AMA-AIH group, only patients who relapsed after immunosuppressive drug withdrawal could be classified with definite AIH. CONCLUSIONS: AMA-AIH shares common features with classical AIH. The diagnosis of AMA-AIH may be swayed by the IAIHG criteria, rendering questionable the applicability of the revised scoring system to this variant form of AIH. PMID- 16704542 TI - 17 Beta-estradiol prevents cytotoxicity from hydrophobic bile acids in HepG2 and WRL-68 cell cultures. AB - BACKGROUND: Epidemiological and clinical studies suggest the possibility that estrogens might have a cytoprotective effect on the liver. The aim of the present study was to test the hypothesis that 17beta-estradiol (E2) prevents hepatocellular damage induced by deoxycholic acid (DCA), a hydrophobic bile acid. METHODS: HepG2 cells were exposed for 24 h to DCA (350 micromol/L). Cell viability, aspartate aminotransferase and lactate dehydrogenase activity and apoptosis were measured as indices of cell toxicity. The effect of DCA was compared to that observed using either a hydrophilic bile acid, ursodeoxycholic acid (UDCA; 100 micromol/L), or E2 at different concentrations (1 nmol/L, 10 nmol/L, 50 nmol/L and 50 micromol/L) or mixtures of E2/DCA or UDCA/DCA. The same experiments were performed using WRL-68 cells that, at variance with HepG2, express a higher level of nuclear estrogen receptor. RESULTS: High concentrations of E2 and UDCA prevented DCA-induced decrease in cell viability, increase in enzyme activity and apoptosis evaluated both by 4',6-diamidino-2-phenylindole dihydrochloride (DAPI) and TdT-mediated dUTP nick-end labeling (TUNEL) assays. In addition, DCA-related apoptosis, assessed by caspase activity, was also prevented by E2 (P < 0.01) in physiological (1-10 nmol/L) doses. The cytoprotective effects of E2 and UDCA was also observed in the WRL-68 cell line. CONCLUSIONS: 17Beta Estradiol prevents DCA-induced cell damage in HepG2 and WRL-68 cell lines to an extent comparable to UDCA. The hypothesis that the protective effect of E2 may be mediated by a mechanism that is nuclear estrogen receptor independent, deserves further verification. PMID- 16704543 TI - Magnetic resonance images of the globus pallidus in patients with idiopathic portal hypertension: a quantitative analysis of the relationship between signal intensity and the grade of portosystemic shunt. AB - BACKGROUND AND AIM: To elucidate a quantitative relationship between hyperintensity of the globus pallidus on T1-weighted magnetic resonance images (MRI) and portosystemic shunt (PSS) in portal hypertension. METHODS: Fifteen patients with idiopathic portal hypertension (IPH) and 44 patients with liver cirrhosis (LC) underwent brain MRI to asses signal intensity at the globus pallidus and Doppler sonography to examine the blood flow volume of PSS. Blood manganese (Mn) levels were examined in 36 patients and neuropsychological tests were performed in 15 patients without overt hepatic encephalopathy. RESULTS: Pallidal hyperintensity on MRI was more prominent in patients with IPH than in patients with LC. There was no correlation between MRI pallidal hyperintensity and the severity of liver dysfunction or hepatic encephalopathy. The grade of hyperintensity correlated well with the grade of PSS. The correlation was stronger in patients with IPH than in patients with LC. The plasma ammonia level and whole blood Mn level significantly correlated with MRI pallidal hyperintensity, but blood Mn level showed a stronger correlation than plasma ammonia. CONCLUSION: Hyperintensity of the globus pallidus on T1-weighted MRI correlated with the development of PSS independent of liver cell function. This brain image should be an index of the grade of PSS rather than a landmark of chronic liver failure. PMID- 16704544 TI - Hemorheology in patients with liver cirrhosis: special emphasis on its relation to severity of esophageal variceal bleeding. AB - BACKGROUND: Patients with decreased blood viscosity are supposed to have a higher risk of bleeding and increased severity of bleeding (severity of bleeding proportional to transmural pressure x area of variceal tear/blood viscosity). However, the hemorheological factors have never been assessed in patients with esophageal variceal bleeding. Therefore, the purpose of the present study was to examine the hemorheological factors in liver cirrhotic patients with special emphasis on the outcome of variceal bleeding. METHODS: Forty-two liver cirrhosis patients with variceal bleeding and another 44 matched patients without bleeding were enrolled. The hemorheological and hemostatic factors of their peripheral blood were examined. The clinical course was under careful surveillance. RESULTS: Patients with poor hepatic reserve (Child B + C vs A) had lower whole blood viscosity (4.34 +/- 0.56 mPa.s vs 5.06 +/- 1.35 mPa.s, P < 0.05), lower hematocrit levels (32.86 +/- 5.97% vs 36.62 +/- 5.44%, P < 0.05), lower platelet counts (79.7 +/- 47.6 x 10(3)/mL vs 108.0 +/- 71.2 x 10(3)/mL, P < 0.05) and prolonged prothrombin time (2.88 +/- 2.33 s vs 1.27 +/- 1.37 s, P < 0.05). Patients with bleeding (vs non-bleeding group) had lower hematocrit levels (31.44 +/- 5.75% vs 36.57 +/- 5.19%, P < 0.01) and lower fibrinogen levels (226.7 +/- 92.7 mg/dL vs 286.4 +/- 111.8 mg/dL, P < 0.05). Patients with bleeding with shock had worse liver cirrhosis (Child A/B/C = 0/5/4 vs 11/18/4, P < 0.05), lower whole blood viscosity (4.01 +/- 0.17 mPa.s vs 4.57 +/- 0.76 mPa.s, P < 0.05), reduced erythrocyte aggregability (2.94 +/- 0.41 vs 3.54 +/- 0.61, P < 0.001), and lower platelet counts (56.22 +/- 17.05 x 10(3)/mL vs 88.87 +/- 38.12 x 10(3)/mL, P < 0.001). The Child-Pugh grade and erythrocyte aggregability were two independent factors associated with bleeding shock. CONCLUSIONS: Whole blood viscosity, hematocrit levels and platelet counts were lower in patients with advanced liver cirrhosis. Advanced liver cirrhosis and reduced erythrocyte aggregability were independent factors for hypovolemic shock in cirrhotic patients with esophageal variceal bleeding. However, the causal relationship between hemorheology and bleeding needs to be clarified in further studies. PMID- 16704545 TI - Hepatobiliary and pancreatic: gallbladder perforation. PMID- 16704546 TI - Gastrointestinal: gastric wall hematoma. PMID- 16704547 TI - Gastrointestinal: mesenteric cystic lymphangioma. PMID- 16704548 TI - Gastrointestinal: midgut malrotation in an adult. PMID- 16704549 TI - Hepatobiliary and pancreatic: spontaneous intrahepatic portosystemic venous shunt. PMID- 16704550 TI - Marked pneumoperitoneum 3 weeks after percutaneous endoscopic gastrostomy. AB - In November 2001, a 29-year-old woman was admitted to the hospital because of dysphagia due to an apallic state caused by cerebral anoxia. Nutritional support was maintained by nasogastric tube feeding for approximately 3 months. For improvement of the body state maintenance and quality of life, a percutaneous endoscopic gastrostomy (PEG) was performed. Three weeks after the PEG, the patient had a wound infection and abdominal distension appeared. Marked pneumoperitoneum was confirmed by radiological examination. No signs or symptoms of peritoneal inflammation developed. A gastrografin study showing that the PEG tube was in the stomach appropriately was checked, and it was noted to be firmly in place without extravasation of contrast. After suspension of the tube feeding and tube opening to decrease intragastric pressure, intravenous hyperalimentation was performed. The pneumoperitoneum resolved within 7 days. Forty days after the PEG, tube feeding was resumed successfully. No recurrence of pneumoperitoneum developed and the patient has remained stable until the present time. The etiology of this finding probably occurs by insufficient fixation of the PEG, causing leakage of air through the gastric wall which enters the free peritoneal space. We recommend that the external binder should be kept 1 cm away from the abdominal skin after the gastrostomy fistula has formed and matured, and periodic rotation of the tube to verify that the internal bumper is free and sufficiently fixed to the gastric wall. In the case of abdominal distension after PEG placement, a X-ray examination and computed tomography (CT) scan with contrast medium would be helpful to ascertain pneumoperitoneum. PMID- 16704551 TI - Successful retrieval of video capsule endoscopy retained at ileal stenosis of Crohn's disease using double-balloon endoscopy. PMID- 16704552 TI - Case of dengue virus infection presenting with acute acalculous cholecystitis. PMID- 16704553 TI - Jejunojejunal intussusception secondary to a jejunal lipoma in an adult. PMID- 16704554 TI - Calcium and vitamin D in the prevention and treatment of osteoporosis - a clinical update. AB - Combined calcium and vitamin D supplementation is an essential component of the management of osteoporosis, supported by a strong scientific rationale. The types of individuals who should receive calcium and vitamin D supplements are those: (i) patients with documented osteoporosis receiving antiresorptive or anabolic treatment; (ii) patients receiving glucocorticoids; and (iii) individuals with or at high risk of calcium and/or vitamin D insufficiencies, in particular older women and men. This article describes the evidence base that supports targeting these groups. Benefits are most apparent when 800 IU day(-1) vitamin D is complemented with a dose of 1000-1200 mg day(-1) elemental calcium. Compliance is also key to optimizing clinical efficacy. PMID- 16704555 TI - Long-term beneficial effects of glipizide treatment on glucose tolerance in subjects with impaired glucose tolerance. AB - AIMS: To assess the efficacy and long-term effects of glipizide treatment on glucose and insulin metabolism in individuals with impaired glucose tolerance (IGT). METHODS: Thirty-seven first-degree relatives of patients with type 2 diabetes fulfilling WHO criteria for IGT were randomized to treatment with either glipizide 2.5 mg once daily or matching placebo for 6 months. A 75 g, 2-h oral (OGTT) and 60 min intravenous glucose tolerance test (IVGTT) were performed at baseline and after 6 months. The subjects were followed up for another 12 months after discontinuation of treatment and a repeat OGTT was performed at 18 months. RESULTS: Thirty-three subjects fulfilled the study. Markers of insulin sensitivity - i.e. fasting insulin and HOMA(IR)-index - improved in the glipizide group (P = 0.04 and 0.02 respectively) as well as HDL cholesterol (P = 0.05) compared with placebo group after 6 months. At 18 months, both fasting and 2 h glucose concentrations were significantly lower in the glipizide group compared with the placebo group (P = 0.04 and 0.03 respectively). The prevalence of type 2 diabetes was 29.4% in the placebo group and 5.9% in the glipizide group at 18 months. This equals an 80% relative risk reduction in the active treatment group. CONCLUSIONS: Short-term treatment with glipizide improves glucose and insulin metabolism in subjects with IGT primarily by improving insulin sensitivity mediated by lowering glucose toxicity, thereby providing the beta cells rest. Larger studies are needed to establish whether these effects are sufficient to prevent progression to manifest type 2 diabetes and associated cardiovascular morbidity in subjects at increased risk of developing type 2 diabetes. PMID- 16704556 TI - A simple linear model for the effect of changes in metabolic risk factors on coronary heart disease. AB - OBJECTIVES: The risk of having one or more metabolic risk factors in the development of coronary heart disease (CHD) is well estimated, but it still remains to be shown which influence any given change in the number of risk factors has on the overall risk of CHD. DESIGN: In this prospective cardiovascular population study, 10 194 participants were examined twice with a 5 year interval for metabolic risk factors such as obesity, hypertension, hypercholesterolaemia and diabetes mellitus. RESULTS: During 14 years of follow up, 1724 incident cases of CHD were identified, 973 in men and 751 in women. The effect of an increase in the number of metabolic risk factors during a 5-year interval on the relative risk of CHD could be determined to be statistically significant for both genders. But only for men, the effect of a decrease in the number of metabolic risk factors was statistically significant during the same time interval. Hence, by statistical analysis, a simple linear model could be constructed for men with two linear trend parameters, one corresponding to the number of metabolic risk factors at the first examination and one corresponding to the change in the number of metabolic risk factors from the first to the second examination. The parameters were 1.50 (1.39-1.63); P < 0.001 and 1.29 (1.18-1.41); P < 0.001, respectively. CONCLUSIONS: This study suggests that it is possible to fit a simple linear model for the effect of changed number of metabolic risk factors on the risk of CHD. PMID- 16704557 TI - Endothelium-dependent vasodilatation in forearm is impaired in stroke patients. AB - OBJECTIVES: To investigate endothelium-dependent vasodilatation (EDV) in stroke patients. DESIGN: Cross-sectional. SETTING: University referral hospital. SUBJECTS: We studied 23 stroke patients (65-75 years old) who did not have atrial fibrillation or Warfarin treatment. Forty-six age- and sex-matched healthy controls and another 32 controls matched also for hypertension and medication were used for comparisons. METHODS. Endothelium-dependent vasodilatation was measured with venous occlusive plethysmography of forearm blood flow during intra brachial infusion of acetylcholine. Endothelium-independent vasodilatation (EIDV) was evaluated by infusion of sodium nitroprusside. RESULTS: Stroke patients showed significantly lower EDV (P < 0.001) compared with healthy controls when measured with acetylcholine-stimulated forearm blood flow. The difference between these groups remained significant also after correction for blood pressure, body mass index, blood glucose and cholesterol. There was also significant difference in EDV between hypertension-matched controls and healthy controls. However, EIDV was significantly reduced in stroke patients (P < 0.01), but not in the hypertensive group, when compared with healthy controls. CONCLUSION: An impaired EDV was seen in both stroke patients and hypertensive-matched controls, while an impaired EIDV was seen in the stroke patients only, suggesting a more severe vasodilatory dysfunction in stroke patients than could be explained by a high blood pressure only. PMID- 16704558 TI - Low testosterone levels are associated with carotid atherosclerosis in men. AB - OBJECTIVE: To study the relationship between endogenous sex hormone levels and intima-media thickness (IMT) of the carotid artery measured by ultrasonography. DESIGN: Population-based cross-sectional study. METHODS: Sex hormone levels measured by immunoassay, anthropometric measurements and IMT was studied in 1482 men aged 25-84 years participating in the 1994-1995 Tromso study. The data were analysed with partial correlation, multiple linear regression and logistic regression analysis. RESULTS: Linear regression models showed that total testosterone and sex hormone-binding globulin levels, but not calculated free testosterone, serum oestradiol or dehydroepiandrosterone sulphate levels were inversely associated with the age-adjusted IMT (P = 0.008 and P < 0.001 respectively). These associations were independent of smoking, physical activity, blood pressure and lipid levels, but were not independent of body mass index (BMI). Excluding men with cardiovascular disease (CVD) did not materially change these results. In a logistic regression model adjusted for the confounding effect of CVD risk factors, men with testosterone levels in the lowest quintile (<9.0 nmol L(-1)) had an independent OR = 1.51 (P = 0.015) of being in the highest IMT quintile. CONCLUSIONS: We found an inverse association between total testosterone levels and IMT of the carotid artery in men that was present also after excluding men with CVD, but was not independent of BMI. The clinical relevance of this, however, is uncertain and needs to be investigated in a clinical setting. PMID- 16704559 TI - Diet and physical activity in relation to overall mortality amongst adult diabetics in a general population cohort. AB - OBJECTIVES: To evaluate the effect on overall and cardiovascular mortality of diet and physical activity amongst diabetic persons. DESIGN: Population-based prospective investigation, from 1993 to mid-2004, in the Greek arm of the European Prospective Investigation into Cancer and Nutrition. SETTING: Volunteers from the general adult Greek population. SUBJECTS: From an original sample of 28 572 volunteers, 1013 were taking at enrolment drugs for diabetes mellitus, had no missing information on the study variables and no comorbidity. Diet was assessed at baseline through a validated questionnaire. Proportional hazards regression was used to assess the relation of dietary factors and physical activity with mortality. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Mortality ratios overall and from cardiovascular causes. RESULTS: During 4579 person-years, 80 deaths have occurred, 46 of which from cardiovascular causes. Physical activity was strongly inversely associated with mortality. Two nutritional variables were significantly associated with diabetic mortality, with hazard ratios for increases of daily intake by one standard deviation being 1.31 for eggs [95% confidence interval (95% CI) 1.07 to 1.60] and 1.82 for saturated lipids (95% CI, 1.14 to 2.90). These two associations were considerably stronger for cardiovascular mortality. Waist-to-height ratio was positively, whereas hip circumference inversely associated with mortality. No significant interactions with gender for any of the study variables were evident. DISCUSSION: Amongst confirmed diabetic persons, increased physical activity is associated with significant reduction of mortality, whereas increased consumption of eggs and saturated fats is associated with significant increase of mortality. Monounsaturated lipids are preferable for diabetic persons. PMID- 16704560 TI - Electrocardiographic and troponin T changes in acute ischaemic stroke. AB - BACKGROUND: The mechanisms explaining morphological electrocardiogram (ECG) changes and increased troponin T (TnT) in acute stroke are unclear. The aims of the present study were to assess the prevalence of ECG and TnT changes in acute ischaemic stroke, to investigate whether ischaemic-like ECG changes correlate to a rise in TnT and to examine whether ECG changes and elevated TnT predict a poor short-time outcome. METHODS: From 2000 to 2002 a total of 279 patients suffering from acute ischaemic stroke were included prospectively in the present study. ECG was carried out at admission and on day 1 in all patients. TnT was analysed at admission and on day 1. RESULTS: The most frequent ECG changes were: prolonged QTc 36.0%, ST depression 24.5%, atrial fibrillation 19.9% and T wave inversion 17.8%. In logistic regression analyses, ST depression and Q waves were significantly associated with a rise in TnT. TnT was elevated (>0.04 microg L( 1)) in 26 patients (9.6%). In logistic regression analyses, a rise in TnT was significantly associated with a poor short-term outcome (modified Rankin scale >3). CONCLUSION: ECG changes are prevalent in acute ischaemic stroke. ST depression and Q waves are related to an increase in TnT, suggesting that these ECG changes may indicate coexisting ischaemic heart disease. A rise in TnT predicts a poor outcome. Patients with acute ischaemic stroke should be offered adequate treatment with secondary prevention and preferably a follow-up with focus on cardiologic as well as neurological aspects. PMID- 16704561 TI - Radiographic measure of aorta calcification is a site-specific predictor of bone loss and fracture risk at the hip. AB - OBJECTIVE: To investigate whether aorta calcification (AC) - a surrogate marker of atherosclerosis - is an independent indicator of low bone mass density (BMD), accelerated bone loss, and risk of future fractures in postmenopausal women. DESIGN: A prospective epidemiological study. Follow-up period was 7.5 years. SETTING: Community-based sample followed by a research institute. SUBJECTS: A total of 2662 generally healthy postmenopausal women with a mean age of 65.0 +/- 7.1 years at baseline. MAIN OUTCOME MEASURES: Annual rate of changes in BMD (DEXA) and AC (X-rays), vertebral fractures (X-rays), hip fractures (questionnaire). RESULTS: Advanced AC at baseline was significantly associated with lower BMD and accelerated bone loss from the proximal femur. In a multivariate logistic regression model, age (OR 1.1, 95% CI 1.0-1.2, P = 0.02), body mass index (BMI; OR 0.9, 95% CI 0.8-1.0, P = 0.03) and the severity of AC (OR 2.3, 95% CI 1.1-4.8, P = 0.03) were independent predictors of hip fractures. Adjusted OR for vertebral fracture was 1.2 (95% CI 1.0-1.5, P = 0.12). CONCLUSIONS: Aorta calcification seems to independently contribute to the development of osteoporosis in the proximal femur. Further studies are needed to clarify whether effective atherosclerosis prevention lowers hip fracture risk. PMID- 16704562 TI - Clinical implications of hypermucoviscosity phenotype in Klebsiella pneumoniae isolates: association with invasive syndrome in patients with community-acquired bacteraemia. AB - BACKGROUND: Klebsiella pneumoniae, a Gram-negative bacillus usually forming glistening mucoid colonies with viscid consistency on the culture plate, is a common pathogen causing various clinical infection patterns. However, little is known about the clinical implications of this mucoid character. OBJECTIVE: The purposes of this study, therefore, were to investigate the frequency of hypermucoviscosity (HV) in bacteraemic isolates of K. pneumoniae, and determine the significance of any association between HV and various clinical manifestations. DESIGN: Retrospective observational study. PATIENTS: Patients diagnosed with K. pneumoniae bacteraemia at a community-based university hospital between June 1999 and June 2001 were enrolled in this analysis. MEASUREMENTS: Clinical data concerning comorbid diseases and infection patterns was collected. K. pneumoniae bacteraemic isolates were examined for the presence of HV using a modified string test. The clinical impact of HV and risk factors for the invasive syndrome were assessed using statistical analysis. Polymerase chain reaction (PCR) was performed to detect magA, a gene related to HV phenotype. RESULTS: Overall, 200 (64.9%) of the 308 cases of K. pneumoniae bacteraemia were community acquired infections. Compared with hospital-acquired K. pneumoniae bacteraemia (HA-KpB), community-acquired K. pneumoniae bacteraemia (CA-KpB) was more likely to be monomicrobial in nature (83.5% vs. 64.8%; P < 0.001) and caused by HV strains (41.5% vs. 14.8%; P < 0.001). The prevalence rate of magA among HV phenotypical K. pneumoniae strains was 24.1%. Patients infected with HV-positive strains were more likely to have the distinctive invasive syndrome (i.e. liver abscess, meningitis, pleural empyaema or endophthalmitis) than those infected with HV-negative variants (37.3% vs. 6.8%; P < 0.001). Multivariate logistic regression analysis, adjusted for age, showed that HV phenotype in K. pneumoniae strains (OR 8.86; 95% CI, 3.70-21.25; P < 0.001) was positively associated with the development of the invasive syndrome in CA-KpB cases. CONCLUSIONS: The HV phenotype of K. pneumoniae bacteraemic isolates was associated with the development of a distinctive invasive syndrome. Identification of the HV phenotype should prompt clinicians to initiate aggressive investigations for invasive diseases. PMID- 16704564 TI - Odor identification accuracy declines following 24 h of sleep deprivation. AB - Brain imaging studies demonstrate that sleep deprivation reduces glucose metabolism and blood flow in the prefrontal cortex, and such reductions are associated with impairments in cognitive functioning. Although some of the greatest metabolic declines occur within the orbitofrontal cortex, little is known about the effects of sleep loss on the types of processes mediated by this region, including emotion, motivation, feeding, and olfaction. The present study tested odor identification accuracy when individuals were well rested and again following 24 h of wakefulness. Relative to rested baseline performance, sleep deprived individuals demonstrated a significant decline in the ability to identify specific odors on the Smell Identification Test. This decrement in olfactory functioning occurred concomitantly with slowed psychomotor speed and increased ratings of self-reported sleepiness. Performance on a task that required complex mental set shifting did not change significantly following sleep deprivation, suggesting that the decrements in odor identification could not be attributed to task difficulty. Finally, while there was no relationship between subjective sleepiness and odor identification at rested baseline, greater subjective sleepiness was associated with better odor identification ability following 24 h of sleep loss. Possible implications of these findings are discussed. PMID- 16704563 TI - Critical aneurysmal dilatation of the thoracic aorta in young adolescents with variant hyperimmunoglobulin E syndrome. AB - The autosomal-dominant (AD) form of the hyperimmunoglobulin E syndrome (HIES) has been described as a multisystem disorder including immune, skeletal and dental abnormalities. Recently, the evaluation of patients from families in which HIES was inherited in a manner more consistent with autosomal-recessive (AR) inheritance, showed that AR-HIES is a clinically distinct disease entity. In addition to classical immunologic findings of AD-HIES, the AR form presents with severe recurrent fungal and viral infections with herpes zoster, herpes simplex and characteristic mollusca contagiosa. Furthermore, cerebral vascular sequelae, including vasculitis, infarction and haemorrhage were noted. In this report, we describe the clinical picture of two patients who showed remarkable resemblance to the description of AR-HIES, but also developed fatal aneurysmal dilatation of the thoracic aorta in adolescence. This finding may further consummate the clinical picture of AR-HIES and emphasize the possibility to develop early aortitis, most likely preceding the critical aneurysm formation at older age. This process should be anticipated during childhood in cases with AR-HIES. PMID- 16704565 TI - Evaluating the awakening criterion in the definition of nightmares: how certain are people in judging whether a nightmare woke them up? AB - There is debate about whether to include in the definition of nightmares a criterion that the imagery or emotions of the nightmare caused the person to wake up. This study investigates whether people believe that they can judge this cause of awakening. 42 participants recorded for 14 nights whether they had a dream, and decided for each dream whether it had or had not woken them. They then rated on a 5-point scale (where 1 = very certain and 5 = very uncertain) how certain they were in their decision of whether or not the dream woke them. Participants' mean certainty was high for decisions that the dream woke them (mean certainty = 1.60), and for very unpleasant dreams this mean certainty that the dream woke them was very high (mean certainty = 1.27). Dreams judged to have caused awakening were found to be more unpleasant than dreams judged not to have caused awakening. Although the inclusion of the awakening criterion did not increase the association of nightmare frequency with anxiety, there may be other advantages in the use of the awakening criterion. PMID- 16704566 TI - Sleep Beliefs Scale (SBS) and circadian typology. AB - The aim of this work was to present the Sleep Beliefs Scale (SBS), a 20-item reviewed version of the Sleep Hygiene Awareness by Lacks and Rotert. We also examined for the first time the influence of circadian typology in sleep beliefs. Voluntary and unpaid psychology students participated in the study (n = 510; 182 men and 328 women), from Italy and Spain, aged between 18 and 33 (22.80 +/- 4.14 years). The mean score of SBS was 13.05 (SD = 3.46; range 2-20) in the total sample, with a distribution positive skewness to high score (correct beliefs) (Z = 1.82; P = 0.003). The internal consistency was good (Cronbach's alpha = 0.714) and factor analysis extracted three factors labelled 'Sleep-incompatible behaviours' (eight items), 'Sleep-wake cycle behaviours' (seven items) and 'Thoughts and attitudes to sleep' (five items). Circadian typology influences the total score and that of the three factors, as well the majority of the items that compose the SBS. The morning-type showed the best scores, the evening-type the worst, and the neither-type the medium scores. Moreover, in the men sample, the differences between circadian typology groups were higher than in the women sample. The SBS showed good psychometric properties; however, further studies in other countries, with clinical and non-student samples, and more aged subjects are needed so as to validate this psychometric instrument. The circadian typology is an individual difference that presented significant relationships with the sleep beliefs, the possibility of the evening-type being a risk factor for a worse sleep hygiene, and the maintenance of sleep problems such as insomnia may all be investigated in depth in future research. PMID- 16704568 TI - Subjective sleepiness and accident risk avoiding the ecological fallacy. AB - The present study of sleepiness and accident risk in a HI-FI car simulator aimed to provide subject-level relative risks (RR) with 95% confidence intervals (CI) for different levels of subjective sleepiness measured with the Karolinska Sleepiness Scale (KSS), 1 = very alert, 9 = very sleepy, fighting sleep, an effort to staying awake. Five male and five female shift workers, mean age 37 years, participated with a 2-h drive (08:00-10:00 hours) in a dynamic high fidelity moving base driving simulator, after a night of work and after a night of sleep. Subjective sleepiness was measured with KSS every 5 min and events of incidents (two wheels outside the right lane), accidents (two wheels off the road or four wheels in opposite lane) and crashes (four wheels off the road) were recorded. The probability of an accident was modelled with a Generalized Linear Mixed Model approach to estimate subject-specific effects, rather than group average effects, to avoid the ecological fallacy. The results showed that sleepiness was strongly related to accident risk. An average subject was estimated at 28.2 times (95% CI RR = 10.7-74.1) increased risk at KSS = 8 and at 185 times (95% CI RR = 42-316) at KSS = 9 compared with KSS = 5. There were large individual differences in event propensity that complicates the prediction of absolute accident risk for individual subjects. PMID- 16704567 TI - Challenging sleep in aging: the effects of 200 mg of caffeine during the evening in young and middle-aged moderate caffeine consumers. AB - The aim of this study was to evaluate the effects of a 200-mg administration of caffeine on polysomnographic sleep variables and quantitative sleep electroencephalography (EEG) in 12 young (20-30 years) and 12 middle-aged (40-60 years) moderate caffeine consumers (one to three cups of coffee per day). All subjects were submitted to both a caffeine (200 mg) and placebo (lactose) condition in a double-blind cross-over design. The conditions were separated by 1 week. Compared with the placebo condition, the evening ingestion of caffeine lengthened sleep latency, reduced sleep efficiency, and decreased sleep duration and amount of stage 2 sleep in both age groups. Caffeine also reduced spectral power in delta frequencies in frontal, central and parietal brain areas, but not in prefrontal (PF) and occipital regions. Moreover, caffeine increased spectral power in beta frequencies in frontal and central brain areas in both age groups. A suppression of spectral power in the PF area in low delta frequencies (0.5-1.00 Hz) and a rise in spectral power in the parietal region in high alpha (10.00 12.00 Hz) and beta frequencies (17.00-21.00, 23.00-25.00, 27.00-29.00 Hz) occurred solely in middle-aged subjects. No such changes were noticeable in young subjects. Generally, caffeine produced similar effects in young and middle-aged subjects. Only a few frequency bins showed more effects of caffeine in middle aged subjects compared with young subjects. Furthermore, sleep EEG results do not entirely support the hypothesis that caffeine fully mimics the effects of a reduction of homeostatic sleep propensity when following a normal sleep-wake cycle. PMID- 16704569 TI - Sleepless night, the moon is bright: longitudinal study of lunar phase and sleep. AB - Popular belief holds that the lunar cycle affects human physiology, behaviour and health. We examined the influence of moon phase on sleep duration in a secondary analysis of a feasibility study of mobile telephone base stations and sleep quality. We studied 31 volunteers (18 women and 13 men, mean age 50 years) from a suburban area of Switzerland longitudinally over 6 weeks, including two full moons. Subjective sleep duration was calculated from sleep diary data. Data were analysed using multiple linear regression models with random effects. Mean sleep duration was 6 h 49 min. Subjective sleep duration varied with the lunar cycle, from 6 h 41 min at full moon to 7 h 00 min at new moon (P < 0.001). Average sleep duration was shortened by 68 min during the week compared with weekends (P < 0.001). Men slept 17 min longer than women (P < 0.001) and sleep duration decreased with age (P < 0.001). There was also evidence that rating of fatigue in the morning was associated with moon phase, with more tiredness (P = 0.027) at full moon. The study was designed for other purposes and the association between lunar cycle and sleep duration will need to be confirmed in further studies. PMID- 16704570 TI - TV exposure associated with sleep disturbances in 5- to 6-year-old children. AB - The aim of this study was to investigate the effects of various forms of TV exposure on the quality of children's sleep. In this randomized population-based survey questionnaires concerning TV viewing, sleep disturbances, and psychiatric symptoms were administered to 321 parents of children aged 5-6 years. Sleep disturbance scores were the main outcome measures. Active TV viewing and passive TV exposure were related to sleeping difficulties, especially sleep-wake transition disorders and overall sleep disturbances. Particularly, passive TV exposure and viewing adult-targeted TV programs were strongly related to sleep disturbances. The association remained significant when socioeconomic status, family income, family conflicts, the father's work schedule, and the child's psychiatric symptoms were controlled statistically. The adjusted odds ratios were 2.91 (95% CI 1.03-8.17) and 3.01 (95% CI 1.13-8.05), respectively. TV viewing and particularly passive TV exposure and viewing adult-targeted programs significantly increase the risk of sleeping difficulties. The results suggest that health-care professionals should be aware of the association between TV exposure and sleep disturbances. PMID- 16704571 TI - Circadian and homeostatic sleep regulation in morningness-eveningness. AB - Morningness-eveningness has been associated with the entrained circadian phase. However, we recently identified morning and evening types having similar circadian phases. In this paper, we compared parameters of slow-wave activity (SWA) decay in non-rapid-eye-movement (NREM) sleep between these two subgroups to test the hypothesis that differences in the dynamics of nocturnal homeostatic sleep pressure could explain differences in sleep timing preference. Twelve morning-type subjects and 12 evening-type subjects with evening types (aged 19-34 years) selected using the Morningness-Eveningness Questionnaire were further classified according to the phase of their dim light melatonin onset (DLMO). The six morning types with the earliest DLMO were compared to the six evening types with the latest DLMO ('extreme' phases), and the six morning types with the latest DLMO were compared to the six evening types with the earliest DLMO ('intermediate' phases). Subjects slept according to their preferred sleep schedule. Spectral activity in four midline derivations (Fz, Cz, Pz, Oz) was calculated in NREM sleep and an exponential decay function was applied on SWA data averaged per sleep cycle. In the subjects with intermediate circadian phases, both initial level and decay rate of SWA in Fz were significantly higher in morning than in evening types. No difference appeared between chronotypes of extreme circadian phases. There was no correlation between individual estimates of SWA decay and DLMO. These results support the hypothesis that chronotype can originate from differences in the dissipation of sleep pressure and that homeostatic and circadian processes influence the sleep schedule preference independently. PMID- 16704572 TI - Use of the oculocardiac reflex to assess vagal reactivity during quiet sleep in neonates. AB - Arousal from sleep can be a protective response to life-threatening stimuli. Hence, faults within state-switching processes may lead to fatal events. To investigate the role of the nervous system during cardiac failure triggered by phasic, vagally mediated stimulation, we analysed autonomic and behavioural reactions in 50 premature neonates during quiet sleep (QS) -- a sleep state characterized by a preponderance of tonic, parasympathetic activity. Bradycardia was induced with a standardized ocular compression test. Neither awakening nor behavioural escape reactions were observed during or after an episode of bradycardia. Eighty-six per cent of the provoked bradycardic episodes induced central apnoea. During QS, the neonates' respiratory response and arousability were found to be time-dependent: when the test was performed early in the QS episode, apnoea was more frequent (94%), and no sleep state change occurred. When ocular compression was performed in the later part of the QS episode, a transition towards active sleep was observed, together with significantly fewer episodes of apnoea (64%). These results indicate that a progressive decrease in the respiratory system's responsiveness to phasic, parasympathetic stimulation occurs during QS, whereas arousability increases. Our study suggests that newborns could be more vulnerable to potentially fatal events during the initial portion of a QS episode. PMID- 16704573 TI - The cardiorespiratory activation response at an arousal from sleep is independent of the level of CO(2). AB - Arousal from sleep is associated with transient cardiorespiratory activation. Traditionally, this response has been understood to be a consequence of state dependent changes in the homeostatic control of ventilation. The hypothesis predicts that the magnitude of ventilatory and cardiac responses at an arousal will be a function of the intensity of concurrent respiratory stimuli (primarily PCO(2)). Alternatively, it has been proposed that increased cardiorespiratory activity is due to reflex activation. This hypothesis predicts that the magnitude of the cardiorespiratory response will be independent of respiratory stimuli. To compare these hypotheses we measured minute ventilation (V(i)), heart rate (HR) and blood pressure (BP) during wakefulness and stage 2 sleep, while manipulating P(et)CO(2). Further, we assessed the magnitude of the response of these variables to an arousal from sleep at the various levels of P(et)CO(2). The subjects were male aged 18-25 years. P(et)CO(2) was manipulated by clamping it at four levels during wakefulness [wake eucapnic, sleep eucapnic (Low), and sleep eucapnic +3 mmHg (Medium) and +6 mmHg (High)] and three levels during sleep (Low, Medium and High). The average number of determinations for each subject at each level was 14 during wakefulness and 25 during sleep. Arousals were required to meet American Sleep Disorders Association criteria and were without body movement. The results indicated that average increases in V(i), HR and BP at arousal from sleep did not significantly differ as a function of the level of P(et)CO(2) present at the time of the arousal (all P > 0.05). Further, the magnitude of the ventilatory response to an arousal was significantly less than the values predicted by the homeostatic hypothesis (P < 0.05). We conclude that, in normal subjects, the cardiorespiratory response to an arousal from sleep is not because of a homeostatic response, but of a reflex activation. PMID- 16704574 TI - Individualized and time-variant model for the functional link between thermoregulation and sleep onset. AB - This study makes use of control system model identification techniques to examine the relationship between thermoregulation and sleep regulation. Specifically, data-based mechanistic (DBM) modelling is used to formulate and experimentally test the hypothesis, put forth by Gilbert et al., that there exists a connection between distal heat loss and sleepiness. Six healthy sleepers each spent three nights and the following day in the sleep laboratory: an adaptation, a cognitive arousal and a neutral testing day. In the cognitive arousal condition, a visit of a television camera crew took place and subjects were asked to be interviewed. During each of the three 25-min driving simulator tasks per day, the distal-to proximal gradient and the electroencephalogram are recorded. It is observed from these experimental data that there exists a feedback connection between thermoregulation and sleep. In addition to providing experimental evidence in support of the Gilbert et al. (2004) hypothesis, the authors propose that the nature of the feedback connection is determined by the nature of sleep/wake state (i.e. NREM sleep versus unwanted sleepiness in active subjects). Besides this, an individualized and time-variant model for the linkage between thermoregulation and sleep onset is presented. This compact model feeds on real-time data regarding distal heat loss and sleepiness and contains a physically meaningful parameter that delivers an individual- and time-depending quantification of a well known biological features in the field of thermoregulation: the thermoregulatory error signal T(hypo)(t)-T(set)(t). A validation of these physical/biological features emphasizes the reliability and power of DBM in describing individual differences related to the sleep process. PMID- 16704575 TI - Adaptive servo-ventilation and deadspace: effects on central sleep apnoea. AB - Central Sleep Apnoea (CSA) occurs commonly in heart failure. Adaptive servo ventilation (ASV) and deadspace (DS) have been shown in research settings to reverse CSA. The likely mechanism for this is the increase of PaCO(2) above the apnoeic threshold. However the role of increasing FiCO(2) on arousability remains unclear. To compare the effects of ASV and DS on sleep and breathing, in particular effects on Arousal Index (ArI), ten male patients with heart failure and CSA were studied during three nights with polysomnography plus measurements of PetCO(2). The order of the interventions control (C), ASV and DS was randomized. ASV and DS caused similar reductions in apnoea-hypopnoea index [(C) 30.0 +/- 6.6, (ASV) 14.0 +/- 3.8, (DS) 15.9 +/- 4.7 e h(-1); both P < 0.05]. However, DS was associated with decreased total sleep time compared with C (P < 0.02) and increased spontaneous ArI compared to C and ASV (both P < 0.01). Only DS was associated with increased DeltaPetCO(2) from resting wakefulness to eupnic sleep [(C) 2.1 +/- 0.9, (ASV) 1.3 +/- 1.0, (DS) 5.6 +/- 0.5 mmHg; P = 0.01]. ASV and DS both stabilized ventilation however DS application also increased sleep fragmentation with negative impacts on sleep architecture. We speculate that this effect is likely to be mediated by increased PetCO(2) and respiratory effort associated with DS application. PMID- 16704576 TI - Sleep-dependent changes in cerebral oxygen consumption in newborn lambs. AB - During rapid-eye-movement (REM) sleep in adult subjects, the cerebral metabolic rate of oxygen consumption (CMRO(2)) is as high as that during wakefulness. We investigated whether CMRO(2) during active sleep is already at the waking level in newborn life, to support the role of active sleep as a state of endogenous brain activation during early postnatal development. Newborn lambs, 2-5 days old (n = 6), were instrumented with electrodes for sleep-state scoring, catheters for blood sample withdrawal and pressure monitoring, and a transit-time ultrasonic blood-flow probe around the superior sagittal sinus. At the age of 19 +/- 3 days, blood samples were obtained simultaneously from the carotid artery and the superior sagittal sinus during uninterrupted epochs of wakefulness, quiet sleep, and active sleep. The arteriovenous difference in blood oxygen concentration was multiplied by cerebral blood flow to determine CMRO(2). CMRO(2) during active sleep (47 +/- 5 micromol min(-1)) was similar to the value in wakefulness (44 +/- 6 micromol min(-1)) and significantly higher than in quiet sleep (39 +/- 5 micromol min(-1), P < 0.05). These data show that active sleep provides newborn lambs with brain activity at a level similar to that in wakefulness in terms of cerebral oxygen metabolism. The high CMRO(2) during active sleep supports its functional role during early postnatal development, when time spent in active sleep is at a lifetime maximum, albeit constituting a metabolic challenge for newborns, because of the impairment of systemic and cerebral vascular regulation in this sleep state. PMID- 16704577 TI - Who is pre-occupied with sleep? A comparison of attention bias in people with psychophysiological insomnia, delayed sleep phase syndrome and good sleepers using the induced change blindness paradigm. AB - Cognitive models of insomnia suggest that selective attention may be involved in maintaining the disorder. However, direct assessment of selective attention is limited. Using the inducing change blindness (ICB) paradigm we aimed to determine whether there is attentional preference for sleep-related stimuli in psychophysiological insomnia (PI) relative to delayed sleep phase syndrome (DSPS) and good sleepers (GS). In the ICB task, a visual scene, comprising both sleep related and neutral stimuli, 'flickers' back and forth with one element (sleep or neutral) of the scene changing between presentations. Therefore, a 2 x 3 totally between-participants design was employed. The dependent variable was the number of flickers it took for the participant to identify the change. Ninety individuals (30 per group) were classified using ICSD-R criteria, self-report diaries and wrist actigraphy. As predicted, PI detected a sleep-related change significantly quicker than DSPS and GS, and significantly quicker than a sleep neutral change. Unexpectedly, DSPS detected a sleep-related change significantly quicker than GS. No other differences were observed between the two controls. These results support the notion that there is an attention bias to sleep stimuli in PI, suggesting that selective attention tasks such as the ICB may be a useful objective index of cognitive arousal in insomnia. The results also suggest that there may be an element of sleep preoccupation associated with DSPS. Results are discussed with reference to other experiments on attentional processing in insomnia. PMID- 16704578 TI - Timing of spontaneous sleep-paralysis episodes. AB - The objective of this prospective naturalistic field study was to determine the distribution of naturally occurring sleep-paralysis (SP) episodes over the course of nocturnal sleep and their relation to bedtimes. Regular SP experiencers (N = 348) who had previously filled out a screening assessment for SP as well as a general sleep survey were recruited. Participants reported, online over the World Wide Web, using a standard reporting form, bedtimes and subsequent latencies of spontaneous episodes of SP occurring in their homes shortly after their occurrence. The distribution of SP episodes over nights was skewed to the first 2 h following bedtime. Just over one quarter of SP episodes occurred within 1 h of bedtime, although episodes were reported throughout the night with a minor mode around the time of normal waking. SP latencies following bedtimes were moderately consistent across episodes and independent of bedtimes. Additionally, profiles of SP latencies validated self-reported hypnagogic, hypnomesic, and hypnopompic SP categories, as occurring near the beginning, middle, and end of the night/sleep period respectively. Results are consistent with the hypothesis that SP timing is controlled by mechanisms initiated at or following sleep onset. These results also suggest that SP, rather than uniquely reflecting anomalous sleep-onset rapid eye movement (REM) periods, may result from failure to maintain sleep during REM periods at any point during the sleep period. On this view, SP may sometimes reflect the maintenance of REM consciousness when waking and SP hallucinations the continuation of dream experiences into waking life. PMID- 16704579 TI - European sleep medicine is airborne. PMID- 16704580 TI - European guidelines for the accreditation of Sleep Medicine Centres. AB - This document describes guidelines for accreditation of Sleep Medicine Centres in Europe. These guidelines are the result of a consensus procedure, in which representatives of the European Sleep Research Society (ESRS) and representatives of different European National Sleep Societies (ENSS) were involved. The information obtained during different rounds of consultation was gathered and processed by the members of the Steering Committee of the ESRS. The scope of the guidelines is to define the characteristics of multidisciplinary Sleep Medicine Centres (SMCs), in terms of requirements regarding staff, operational procedures and logistic facilities. Accreditation of SMCs is proposed to be the responsibility of the individual ENSS. The Accreditation Guidelines may thus be considered an instrument for the national societies to develop new or standardize existing accreditation questionnaires, as well as procedures for visiting the site, drafting the accreditation report, and finally, granting the accreditation. The Accreditation Guidelines are meant to be a line of action, that ideally should be followed as close as possible, but that may be subject to certain exceptions, depending on local customs or regulations. PMID- 16704582 TI - Transfer of brain injured patients - time for a change? PMID- 16704583 TI - Effect of spinal flexion on the conus medullaris: a case series using magnetic resonance imaging. AB - Anatomy textbooks state that the conus medullaris moves cephalad when the vertebral column is flexed. This could confer protection against spinal cord damage during dural puncture, but has not been demonstrated in vivo. We therefore imaged the spine of 10 volunteers using magnetic resonance imaging to determine if such movement occurs with the spine in the neutral and flexed positions. The position of the conus medullaris in relation to the superior endplate of the L1 vertebra was determined. On spinal flexion, the conus medullaris moved cephalad in three subjects and caudad in three subjects, with no change in the remaining four. The median overall movement (95% CI [range]) was 0 mm (4 mm caudad to 1 mm cephalad [3 mm caudad to 1 mm cephalad]; p = 1.0). Whilst spinal flexion may facilitate needle insertion during dural puncture, it is unlikely to confer extra protection against spinal cord damage. PMID- 16704584 TI - High concentration potassium permanganate eliminates protein and particle contamination of the reusable Classic laryngeal mask airway. AB - In this three-stage study, we test the hypothesis that supplementary cleaning with potassium permanganate > or =4 mg.l(-1) eliminates protein and particle contamination from the reusable Classic laryngeal mask airway. The first stage involved supplementary cleaning of 70 1 x 1 cm segments from deliberately contaminated laryngeal mask airways using potassium permanganate at 0, 2, 4, 8, 16, 32 and 64 mg.l(-1) and testing for protein staining. This showed that the lowest concentration required to eliminate protein contamination was 8 mg.l(-1). The second stage involved supplementary cleaning of 50 used laryngeal mask airways with either potassium permanganate 8 mg.l(-1) or saline and testing for protein staining. This showed that protein contamination was lower in the potassium permanganate group (p < 0.00001): all laryngeal mask airways in the control group and none in the potassium permanganate group were contaminated. The third stage involved scanning electron microscopic examination of 1 x 1 cm segments from three laryngeal mask airways used in the control group, three from the potassium permanganate group, plus three brand new laryngeal mask airways. The mean density of > or =1 mum surface particles was lower in the potassium permanganate 8 mg.l(-1) than the control group (21 vs. 121 .cm(-2), p < 0.0001) and was similar to brand new laryngeal mask airways (24 .cm(-2)). We conclude that supplementary cleaning with potassium permanganate 8 mg.l(-1) eliminates protein deposits from reusable laryngeal mask airways and reduces particle contamination to similar levels to brand new laryngeal mask airways. PMID- 16704585 TI - Perivascular axillary brachial plexus block and patient positioning: the influence of a lateral, head-down position. AB - The aim of this study was to examine the effect of a 20 degrees Trendelenburg position on the blockade of nerves that exit the brachial plexus proximally in patients undergoing single-injection axillary brachial plexus block. After a pilot study of eight cadavers suggested that a head-down and lateral position would encourage the proximal spread of local anaesthetic, 72 patients undergoing elective surgery were divided into two equal groups: a Supine group and a Modified Position group (lateral position, 20 degrees head-down tilt). Patients were left in the allocated position for 30 min after an axillary block had been performed with alkalinised mepivacaine 1% 49.5 ml. Sensory and motor blockade evaluation showed that there was a significantly higher proportion of axillary nerve (76% vs. 0%, p < 0.001), thoracodorsal nerve (86% vs. 0%, p < 0.001) and subscapular nerve (89% vs. 0%, p < 0.001) blockade in the Modified Position group. Sensory block of the radial nerve was also improved by the modified position (100% vs. 86%, p < 0.05). PMID- 16704586 TI - Inhalation induction using sevoflurane in children: the single-breath vital capacity technique compared to the tidal volume technique*. AB - The single-breath vital capacity technique is suitable for inhalation induction of anaesthesia, using sevoflurane in children aged > 5 years. The purpose of this randomised trial was to compare the single breath vital capacity technique with the conventional tidal volume technique. Seventy- three ASA 1 or 2 children were instructed during the pre-operative visit in the vital capacity technique. The main criterion measured was time to loss of the eyelash reflex. Induction was performed using a circle-absorber breathing circuit primed with sevoflurane 7% in 50% nitrous oxide/oxygen with 6 l.min(-1) fresh gas flow. Time required for induction, haemodynamic changes, airway tolerance and side-effects were recorded. The children's opinion on the technique used was scored using a visual analogue scale (0-100) and a Smiley scale (0-10). The time to loss of the eyelash reflex was found to be reduced in the vital capacity group compared to the tidal volume group. The time to central myosis, to achieve bispectral index values 60 and 40, haemodynamic changes, respiratory events and side-effect incidences were similar in both groups. However, we found that the vital capacity technique was preferred by the children to the tidal volume technique. PMID- 16704587 TI - The effects of brief sevoflurane-nitrous oxide anaesthesia upon children's postoperative cognition and behaviour. AB - This study assessed the effects of brief sevoflurane-nitrous oxide anaesthesia on children's postoperative cognition, behaviour and physical morbidity. Forty-eight children aged 5-10 years undergoing anaesthesia without premedication for multiple dental extractions, and 48 control children, performed tests of choice reaction time, attention, psychomotor co-ordination and memory pre-operatively (baseline), prior to discharge and at 48 h (anaesthesia group only). Physical and psychological morbidity were recorded at 1 week. Mean choice reaction time and psychomotor co-ordination were significantly impaired postoperatively but had recovered at 48 h. However, measures of performance variability suggested the presence of residual impairment. Profound retrograde amnesia affected postoperative and 48-h recall of pictorial stimuli presented prior to anaesthesia, but recognition memory was unimpaired. Attention-seeking, tantrums, crying and nightmares were occurring more frequently in some 8-20% of children 1 week after the procedure. PMID- 16704588 TI - From a radial operating theatre to a self-contained operating table. AB - Equipment congestion and a disarray of wires, tubes and lines (the spaghetti syndrome) is a common scenario in operating theatres. The radial arrangement of input and output signals and their interconnecting lines has been identified as the main source of clutter and congestion in this environment. Our aim was to present a comprehensive design concept for reducing electrical and physical clutter in the operating theatre. Data were collected from different operating theatres, including identification and sorting of equipment, cables, tubes and lines according to the direction and the features of the transmitted information and materials. We suggest a concept of a self-contained, 'built-in' operating table as a design solution for avoiding the clutter and congestion caused by the radial configuration. The operating table will function as a central integrated unit for management of the entire process of patient flow and control of supply systems and environmental conditions. PMID- 16704589 TI - The Robinson-Hooper connection*. AB - The search for a connection between James Robinson, the dental surgeon who on 19 December 1846 administered the first general anaesthetic in England by the inhalation of the vapour of ether, and William Hooper, the pharmaceutical chemist who produced the best publicised and most widely used of the early commercial ether vaporisers, has revealed hitherto unrecognised aspects of the early history of general anaesthesia. PMID- 16704590 TI - An objective assessment of nerve stimulators used for peripheral nerve blockade*. AB - Nerve stimulation is considered by many to be the current 'gold standard' for locating peripheral nerves. Thirteen nerve stimulators were subjected to a battery of tests performed by two independent observers under standardised conditions using a digital oscilloscope, a calibrated resistance and a novel scoring system. Individual scores were assigned for the signal waveform, current intensity, impulse duration, maximum load output and functionality of each nerve stimulator; the maximum score achievable was 20. A score of > or = 15/20 was achieved by six nerve stimulators: Stimuplex HNS 12 (B Braun); MultiStim Vario (Pajunk); Plexival (Medival, Vygon); MultiStim Sensor (Pajunk); Plexygon (Aryon, Vygon); Stimuplex HNS 11 (B Braun). Seven nerve stimulators scored < 15/20. Anaesthetists should be aware of the limitations of the nerve stimulator being used. Standardisation of features is desirable when manufacturing nerve stimulators. PMID- 16704591 TI - Surgical vs wire-guided cricothyroidotomy: a randomised crossover study of cuffed and uncuffed tracheal tube insertion. AB - Using an airway mannequin and artificial lung model, we compared surgical cricothyroidotomy with a 6.0-mm cuffed Portex tracheostomy tube with wire-guided cricothyroidotomy using a 5.0-mm cuffed Melker or 6.0-mm uncuffed Melker tube. The trial was carried out by 27 anaesthetists using a randomised, crossover design. Surgical cricothyroidotomy proved significantly faster (mean (SD) time to first breath 44.3 (12.5) s for Portex surgical, 87.2 (21.6) s for cuffed Melker, 87.8 (19.2) s for uncuffed Melker, p < 0.001). With a standardised ventilator model, the cuffed tubes provided more effective ventilation (mean (SD) tidal volume 446 (41) ml Portex, 436 (52) ml cuffed Melker, 19 (5) ml uncuffed Melker, p < 0.001). Fourteen of the participants preferred the wire-guided system. We conclude that, in this model, a cuffed device is preferable when cricothyroidotomy is needed. In addition, the surgical method is quicker than a wire-guided approach. PMID- 16704592 TI - Simulated clinical evaluation of four fluid warming devices*. AB - The fluid warming capabilities of the Bair Hugger, Hotline, Standard Ranger and Fluido devices were evaluated in the laboratory with gravity flow via a 14G cannula (1 m head of fluid) and with the fluid bag pressurised to 300 mmHg. The resulting flows (70-450 ml.min(-1)) were recorded. At a room temperature of 22 degrees C, no device warmed the fluid to 37 degrees C. However, the Standard Ranger with gravity flow and the Fluido with both gravity and pressurised flow achieved 35 degrees C, whereas the Bair Hugger and Hotline with both gravity and pressurised flow, and the Standard Ranger with pressurised flow, achieved only 24 31 degrees C. However, from the way delivered temperatures changed with flow, we calculated that all four devices would achieve close to 37 degrees C at the flows specified by the manufacturers: 17, 83, 150 and 800 ml.min(-1) for the Bair Hugger, Hotline, Standard Ranger and Fluido, respectively. PMID- 16704593 TI - Evaluation of four airway training manikins as simulators for inserting the LMA Classic*. AB - Airway manikins have traditionally been used for teaching mask ventilation and tracheal intubation. There is an increasing need to use manikins for training in procedures such as insertion of the laryngeal mask airway. We have assessed four new airway training manikins (latest versions of the Airway Trainer (Laerdal, Norway), Airway Management Trainer (Ambu, UK), 'Bill 1' (VBM, Germany) and Airsim (Trucorp, Ireland)) as simulators for insertion of the LMA Classic laryngeal mask airway. Twenty volunteer anaesthetists inserted a size-4 laryngeal mask airway five times into each of the four manikins, in random order. Each insertion was assessed using objective and subjective tests. Subjective assessment varied widely but overall assessment indicated that the Airway Management Trainer was the poorest simulator for insertion of the laryngeal mask airway. The 'Bill 1' and Airsim manikins performed best as simulators for insertion of the laryngeal mask airway, although realistic ventilation with 'Bill 1' was not possible. PMID- 16704594 TI - Hyperpyrexia and prolonged postoperative disorientation following methylene blue infusion during parathyroidectomy. AB - A 65-year-old man underwent parathyroidectomy for hyperparathyroidism secondary to renal failure. Intra-operatively he received methylene blue infusion (7.5 mg.kg(-1), a total of 650 mg in 500 ml 0.9% sodium chloride) for visualisation of parathyroid glands. At the end of surgery, following extubation he developed agitation, intense shivering and hyperpyrexia, and his level of consciousness decreased to a Glasgow Coma score of 7. The differential diagnoses included methylene blue toxicity or malignant hyperpyrexia. His lungs were ventilated, and intravenous dantrolene was administered to control hyperpyrexia. Haemodialysis was started to remove the methylene blue dye. We review the literature on the pharmacological actions of methylene blue, and discuss the differential diagnosis and management of this patient. PMID- 16704595 TI - Chylothorax as a complication of innominate vein thrombosis induced by a peripherally inserted central catheter. AB - A 5-month-old boy with hypoplastic left heart syndrome developed chylothorax and oedema of the left upper arm more than 2 months after recovering from a Norwood stage II operation. Venography showed occlusion of the innominate vein with abundant collaterals crossing the midline to join the right internal jugular vein. The occlusion was caused by a peripherally inserted central catheter under systemic heparinisation. This case highlights the importance of a patent venous pathway, especially in patients with a cavopulmonary connection in the upper extremities. PMID- 16704596 TI - Endobronchial tubes - a case for re-evaluation. AB - An endobronchial tube (Macintosh-Leatherdale) was used to secure the airway for a tracheal resection and end-to-end anastomosis. This lung separation device enabled insertion of both a fibreoptic bronchoscope and a tube exchange catheter. These were required after the trachea was transected and re-anastomosis proved surgically difficult. The airway exchange catheter allowed for jet ventilation and later a tube change when an emergency occurred. Options and management issues for tracheal surgery and lung separators are discussed. A case is made for a re evaluation of endobronchial tubes both as a useful conduit for modern airway instruments and as an alternative to small double-lumen tubes for the increasing population of obese patients weighing > 100 kg, requiring thoracic surgery. PMID- 16704597 TI - Epidural anaesthesia as a complication of attempted brachial plexus blockade using the posterior approach. AB - We report a case of accidental epidural anaesthesia as a complication of attempted brachial plexus blockade using a posterior approach in a 31-year-old man scheduled to undergo elective shoulder surgery. The block was inserted with the patient in the lateral position before induction of general anaesthesia. On emergence from anaesthesia, the patient could breathe but could not move his arms. He had no pain sensation from the fifth cervical dermatome to the third thoracic dermatome bilaterally; this resolved 8 h after surgery and he was discharged well 2 days later. Although proponents of the posterior approach to the brachial plexus claim that its use is associated with a lower incidence of significant complications, this case proves that the technique is not devoid of potentially serious complications. PMID- 16704598 TI - Major colorectal surgery in a patient with cold agglutinin disease. AB - We present the case of a 62-year-old man with severe cold agglutinin disease who underwent major colorectal surgery. Cold agglutinin disease is a condition in which auto-antibodies, usually immunoglobulin M, cause red blood cell agglutination at decreased body temperature. Haemolysis may result. Agglutination results in impaired perfusion, resulting in symptomatic Raynaud's phenomenon and acrocyanosis. Haemolysis can result in anaemia and thrombotic events caused by microvascular occlusion, in addition to haemoglobinuria and renal failure. Peri operative hypothermia is common in all patients and may be associated with significant morbidity, but is potentially catastrophic in a patient suffering from cold agglutinin disease. PMID- 16704599 TI - Spinal myoclonus following combined spinal-epidural anaesthesia for Caesarean section. AB - A nulliparous woman presented with pre-eclampsia at 39 weeks' gestation. A combined spinal-epidural anaesthesia was employed for Caesarean section but the spinal component produced no discernible block, so the epidural was topped up with 20 ml ropivacaine 0.75% without problem and surgery was uneventful. A week after delivery she developed twitching of her legs and opisthotonus, that was initially thought to be eclampsia but was subsequently diagnosed as spinal myoclonus. She was treated with oral carbamazepine and diazepam, with improvement over the next 4 days, and discharged home a week later taking oral carbidopa and levodopa. Her symptoms resolved completely 6 months after the initial event. PMID- 16704600 TI - Use of the fibreoptic intubating LMA-CTrach in two patients with difficult airways. AB - We report two patients with difficult airways who underwent tracheal intubation using the new fibreoptic intubating Laryngeal Mask Airway CTrach. The imaging technology of the LMA-CTrach was decisive in the management of these two patients. The first patient had lingual tonsillar hyperplasia, and an omega shaped retroflexed epiglottis. The second patient had a C2-occipital fusion and was completely unable to extend her head. Given the anatomical difficulties encountered, it was likely that intubation would have been difficult or impossible through the LMA-Fastrach. The aim of this report is to describe the successful use of this new intubating laryngeal mask airway in these two challenging patients. PMID- 16704601 TI - Brush up your diagnosis. PMID- 16704602 TI - Failed intubation in obstetric anaesthesia. PMID- 16704605 TI - Lipid emulsion to treat overdose of local anaesthetic. PMID- 16704606 TI - Are all single-use laryngeal masks the same? PMID- 16704607 TI - Criterion contamination when using the Hospital Anxiety and Depression Scale. PMID- 16704611 TI - Pressure ulcers in intensive care. PMID- 16704612 TI - Routine pre-oxygenation. PMID- 16704613 TI - Hypothermia and spinal anaesthesia. PMID- 16704614 TI - A combined cannula-over- needle and wire-guided technique for cricothyroidotomy. PMID- 16704615 TI - Preventing extravasation from central venous catheters. PMID- 16704616 TI - Risk of nerve damage from vascular access catheters. PMID- 16704617 TI - Ultrasound to detect incorrect guidewire positioning during subclavian line insertion. PMID- 16704618 TI - Mobile phone use in hospitals. PMID- 16704619 TI - A response to 'Generic machine checks'. PMID- 16704621 TI - A response to 'Evaluation of capsaicin ointment at the Korean hand acupressure point K-D2 for prevention of postoperative nausea and vomiting'. PMID- 16704622 TI - A response to 'Airway rescue in acute upper airway obstruction using a ProSeal Laryngeal mask airway and an Aintree Catheter: a review of the ProSeal Laryngeal mask airway in the management of the difficult airway'. PMID- 16704623 TI - A response to 'Survival after massive bleeding into the airway in a patient at risk from "can't intubate, can't ventilate"'. PMID- 16704625 TI - A response to 'The nuclear power industry as an alternative analogy for safety in anaesthesia and a novel approach for the conceptualisation of safety goals'. PMID- 16704626 TI - A response to 'Autotriggering of pressure support ventilation during general anaesthesia'. PMID- 16704627 TI - A response to 'Life-threatening pulmonary hypertension and right ventricular failure complicating calcium and phosphate replacement in the intensive care unit'. PMID- 16704628 TI - A response to 'Use of carbon dioxide extubation'. PMID- 16704631 TI - How safe is patch testing to PPD? PMID- 16704632 TI - Updates from the British Association of Dermatologists 85th annual meeting, 5-8 July 2005, Glasgow, U.K. AB - The conference highlighted the progress made in understanding recent biological, epidemiological and therapeutic advances in dermatology. Here we provide a synopsis of the main research and clinical findings presented at the meeting of the British Association of Dermatologists (BAD) held during 5-8 July 2005, in Glasgow, U.K., drawing attention to the most important advances and summaries. The BAD meeting was held at the Scottish Exhibition and Conference Centre, Glasgow (Fig. 1). The annual dinner was held in the wonderful setting of Stirling Castle, with Dr Robin Graham-Brown as host. PMID- 16704633 TI - Do we need a revised public health policy on sun exposure? PMID- 16704634 TI - A new government policy is needed for sunlight and vitamin D. PMID- 16704635 TI - Expression of CD90 on keratinocyte stem/progenitor cells. AB - BACKGROUND: The identification and purification of keratinocyte stem cells (KSCs) that are capable of self-renewal and maintenance of differentiating cell populations could contribute both to our understanding of the biology of these cells, and to significant clinical applications, such as the culturing of keratinocytes for transplantation to severe burn wounds. Here, we report the detection of CD90(+) cells in cultured normal human epidermal keratinocytes and adult skin. OBJECTIVES: To investigate the biological function of CD90(+) and CD90(-) keratinocytes. METHODS: CD90(+) and CD90(-) keratinocytes were purified from adult skin and cultured keratinocytes using fluorescent activated cell sorting, and their biological abilities were analysed using both in vitro and in vivo assays. RESULTS: Flow cytometry (FCM) analysis identified approximately 18% of post-primary neonatal keratinocytes as CD90(+). However, during expansion of the culture, the expression level of CD90 rapidly decreased to about 2.5% at passage 10, while most of the keratinocytes maintained expression of alpha6 integrin. Purified CD90(+) keratinocytes demonstrated a sixfold higher cell growth rate than CD90(-) cells and the ability to form large (over 3 mm in diameter) colonies. We then quantitatively evaluated both populations using a previously described in vivo human epidermal cyst formation assay. Enhanced green fluorescent protein (EGFP)-labelled CD90(+) or CD90(-) keratinocytes were subcutaneously injected into nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice. Six weeks after transplantation, EGFP(+) cell clusters in human epidermal cysts were evaluated using image analysis software. EGFP(+) cell cluster areas in the basal layer, derived from EGFP(+) CD90(+) cells, were eightfold larger than clusters of EGFP(+) CD90(-) cells. Furthermore, immunohistochemical staining and FCM analysis indicated that CD90 was expressed in most of the basal layer of the normal human epidermis. CONCLUSIONS: These results indicated that CD90 is a useful marker for the detection of human KSC enriched populations in cultured human keratinocytes. PMID- 16704637 TI - Sentinel lymph node status in melanoma: a valuable prognostic factor? AB - BACKGROUND: Sentinel lymph node (SLN) biopsy is advocated as the standard of care for patients with primary melanoma. It is a procedure with few side-effects and provides valuable staging information about the regional lymphatics. OBJECTIVES: To investigate the prognostic value of SLN biopsy and to compare it with that of other known risk factors in primary melanoma. METHODS: One hundred and forty-nine patients with primary melanomas (tumour thickness >1.0 mm) underwent SLN biopsy between May 1998 and April 2004 at our department. This report summarizes the follow-up data of this cohort until October 2004. RESULTS: SLN biopsies of 49 of 149 patients (33%) revealed micrometastatic disease. Of all clinical and histological criteria, only the clinical type of primary melanoma (11 of 19 patients with acrolentiginous melanomas) and the Clark level were predictive for SLN positivity. Progression was observed in 22 patients (15%). It was significantly associated with ulceration of the primary tumour, tumour thickness, clinical type and localization of the primary tumour, female sex and older age. In contrast, SLN positivity was not significantly associated with a higher risk of progression (eight of 49 SLN-positive vs. 14 of 100 SLN-negative patients; P = 0.807). Twelve of 149 patients (8%) died because of melanoma in the follow-up period. Significant criteria for death were ulceration of the tumour, clinical type and localization of the primary tumour, but not SLN positivity. CONCLUSIONS: A high percentage of positive SLNs was observed in the patients with melanoma in our study (33%). The fractions of patients both with progressive disease and with tumour-related death were not significantly higher in patients with positive SLN than in those with negative SLN. We therefore conclude that the SLN status is not a reliable prognostic factor for progression of melanoma. PMID- 16704636 TI - Dopamine-induced apoptosis in human melanocytes involves generation of reactive oxygen species. AB - BACKGROUND: Previous studies have shown that significant increases in urinary and plasma levels of several monoamines and their metabolites characterize the onset of vitiligo and its progression. Recently, both epidermal keratinocytes and melanocytes were found to have the capacity for the biosynthesis of several catecholamines and serotonin. Some monoamines and their metabolites can induce apoptosis and cytotoxicity in neural cells. However, no previous report has investigated the potential role of these monoamines in inducing apoptosis or cytotoxicity in melanocytes. OBJECTIVES: To study the effects of dopamine (DA), norepinephrine (NE), epinephrine (EP), and serotonin (5-HT) on melanocyte cytotoxicity and apoptosis. METHODS: Primary cultures of normal human melanocytes established from the foreskins of normal individuals were treated with different concentrations of DA, NE, EP and 5-HT for 5 and 7 days. Cell viability was measured by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Melanocyte apoptosis was evaluated by morphological examination and flow cytometric analysis. We also measured the generation of reactive oxygen species (ROS) after DA treatment. RESULTS: Among the four monoamines used in this study, only DA had an effect, dose-dependently decreasing the melanocyte viability at concentrations ranging from 0.01 to 100 micromol L(-1) (0.1 and 1 micromol L(-1), P < 0.05; 10 micromol L(-1), P < 0.01). In addition, DA-induced melanocyte apoptosis was evidenced by the increased percentage of sub-G1 cells from 7.71 +/- 0.28% (control) to 12.22 +/- 1.05% (0.1 micromol L(-1) DA) (P < 0.005), and treatment with the antioxidant N-acetylcysteine (NAC) reversed this apoptotic effect. DA treatment led to the generation of ROS, which could be prevented by pretreatment with NAC. CONCLUSIONS: DA can induce melanocyte apoptosis, which might be related to the generation of ROS. This novel effect might play an important role in the development or progression of vitiligo, which is currently viewed as a disease process closely related to melanocyte apoptosis. PMID- 16704638 TI - Dermatological manifestations of autoimmune polyendocrinopathy-candidiasis ectodermal dystrophy syndrome. AB - BACKGROUND: Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy syndrome (APECED; OMIM 240300) is a rare autosomal recessive disorder defined by a variable combination of endocrine failure, chronic mucocutaneous candidiasis (CMC), and dystrophy of the dental enamel and nails. APECED is caused by mutations in the autoimmune regulator gene (AIRE). Alopecia areata (AA) and vitiligo are diseases with autoimmune pathogeneses, and have been recognized as part of the APECED complex. There are rare reports of other cutaneous manifestations. OBJECTIVES: We sought to delineate the dermatological features of APECED in an Irish case series with emphasis on timing of their appearance and association with disease severity. Furthermore, we looked for evidence of genotype: phenotype correlation. Finally, we wanted to determine if the ectodermal changes described represent a primary ectodermal dysplasia or whether the ectodermal manifestations are secondary phenomena. METHODS: Irish patients with APECED were invited to attend a multidisciplinary clinic (Dermatology, Endocrinology, Dentistry and Ophthalmology) held in Our Lady's Hospital for Sick Children, Dublin. Clinical data were compiled from case notes and questionnaires. All patients had a detailed cutaneous examination. Blood samples were obtained for mutational analysis. RESULTS: Eighteen patients (seven males and 11 females) from 15 families were interviewed and examined. The mean age at diagnosis was 6 years (range 8 months-18 years). All patients had evidence of CMC, 13 (72%) had candidal onychomycosis or paronychia, six (33%) had AA and two had vitiligo. In the case of two patients the diagnosis was made on recognition of dermatological manifestations and confirmed by mutational analysis. Both patients developed Addison's disease on follow-up. CMC was an early feature, often predating diagnosis (10 of 18). AA and vitiligo presented later, and may reflect more severe disease in these cases. There was no correlation between the AIRE mutations identified on mutational analysis and the clinical presentation. We found no evidence of an isolated nail dystrophy or features consistent with a primary ectodermal dysplasia. CONCLUSIONS: APECED is a rare but complex and potentially life-threatening autoimmune disease. CMC is a common and early feature; diagnosis at this stage may pre-empt life-threatening endocrinological crises. It is important for dermatologists to be aware of this association as they are likely to be the earliest clinicians who encounter these children. AA and vitiligo in our series occurred in the setting of established disease. The term "ectodermal dystrophy" is misleading as the ectodermal features described in our series and in the literature are most likely to be secondary phenomena. PMID- 16704639 TI - The dermal stem cell factor and c-kit are overexpressed in melasma. AB - BACKGROUND: The pathogenesis of melasma has not yet been clearly demonstrated. We tried to determine whether the stem cell factor (SCF) and its receptor c-kit are involved in the mechanism of hyperpigmentation of melasma because this factor is highly implicated in the stimulation of melanocyte function in vitro and in vivo. OBJECTIVES: The present study was conducted to investigate the expression of SCF and c-kit on the lesions of melasma compared with nonlesional skin. PATIENTS/METHODS: Skin samples were obtained from lesional and nonlesional facial skin of 60 Korean women with melasma. Immunohistochemistry and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis was performed to determine the expression of SCF and c-kit in melasma. RESULTS: The expression of SCF was significantly increased at the lesional dermis compared with nonlesional dermis. However, there was no significant difference in the expression of SCF in lesional and nonlesional epidermis. The expression of c-kit was significantly increased at lesional epidermis compared with nonlesional skin. RT-PCR of SCF and c-kit mRNAs demonstrated increased expression of both types of transcripts in the lesional skin compared with nonlesional skin. CONCLUSIONS: These results suggest that the increased expression of SCF in the dermis and of c-kit in the epidermis play an important role in the mechanism of hyperpigmentation in melasma. PMID- 16704640 TI - Transepidermal water loss, serum IgE and beta-endorphin as important and independent biological markers for development of itch intensity in atopic dermatitis. AB - BACKGROUND: Although itch is the predominant symptom of atopic dermatitis (AD), it is poorly characterized and subjective. The objective assessment of itch intensity is important for treatment and follow-up in patients with AD. OBJECTIVES: To determine what objective clinical parameter(s) could be used as biomarker(s) for itch intensity in patients with AD. METHODS: This is a retrospective and cross-sectional study. Seventy-five patients, aged 7 months-49 years with equal sex ratio, were enrolled in 2000 according to criteria proposed by Hanifin and Rajka. Thirty-five age- and sex-matched subjects who visited the dermatological clinic but were otherwise healthy served as controls. Subjective itch intensity was divided into four grades of severity. Disease severity was measured by SCORAD index, which also includes itch intensity as part of the measurement. Transepidermal water loss (TEWL) and skin surface pH were measured by noninvasive methods in clinically normal skin on the forearm. Serum beta endorphin and vasoactive intestinal peptide (VIP) were determined by radioimmunoassay. Ordinal logistic regression was used to assess the trend of the subjective itch intensity and SCORAD index by serum IgE, beta-endorphin, VIP, TEWL and skin pH. RESULTS: There were significant trends for itch intensity with IgE, beta-endorphin and TEWL. After adjustment for sex, age and other variables, the odds ratio (OR) for itch intensity by log IgE, beta-endorphin and TEWL was 2.103 [95% confidence interval (CI) 1.222-3.618], 1.100 (95% CI 1.005-1.203) and 1.081 (95% CI 1.009-1.158), respectively. The OR for disease severity by log IgE, beta-endorphin and TEWL was 2.250 (95% CI 1.149-4.407), 1.156 (95% CI 1.086 1.231) and 1.071 (95% CI 0.971-1.182), respectively. In contrast, there was no association between serum VIP concentration and itch intensity. CONCLUSIONS: Beta endorphin and IgE are both useful biomarkers for itch and disease severity in patients with AD, while TEWL is a good biomarker for itch intensity. These biomarkers provide a way to assess the itch intensity in patients with AD. PMID- 16704641 TI - Dermoscopic findings in pyogenic granuloma. AB - BACKGROUND: To the best of our knowledge, no specific dermoscopic criteria have been described in the medical literature for the diagnosis of pyogenic granuloma. OBJECTIVES: To evaluate the morphological findings of pyogenic granuloma under dermoscopic observation. METHODS: Dermoscopic examination (using the DermLite Foto; 3Gen, LLC, Dana Point, CA, U.S.A.) of 13 patients with pyogenic granulomas was performed to evaluate specific dermoscopic criteria. RESULTS: The most frequently occurring dermoscopic features were found to be: reddish homogeneous area (92%), white collarette (85%), "white rail" lines that intersect the lesion (31%) and ulceration (46%). The results of our study reveal that the absence of specific dermoscopic criteria for other skin tumours and a reddish homogeneous area surrounded by a white collarette are the most frequent dermoscopic pattern in pyogenic granulomas (85%). CONCLUSIONS: Dermoscopy is a useful tool for improving the recognition of pyogenic granuloma. PMID- 16704642 TI - Significance of interleukin-16, macrophage-derived chemokine, eosinophil cationic protein and soluble E-selectin in reflecting disease activity of atopic dermatitis--from laboratory parameters to clinical scores. AB - BACKGROUND: The search for the ideal clinical score reflecting atopic dermatitis (AD) severity has developed in parallel with unveiling key events in disease pathogenesis and finding laboratory parameters for monitoring disease activity. A major difficulty in assessing the relevance of reported serum markers of AD severity is the use of nonvalidated referent tools, which compromises comparison of results across studies. OBJECTIVES: The aim of our study was to compare the significance of serum levels of interleukin (IL)-16, macrophage-derived chemokine (MDC), soluble E-selectin (sE-selectin) and eosinophil cationic protein (ECP) in reflecting AD severity and identify the most relevant parameter for monitoring the course of disease. Serum levels were tested against the same referent severity score in the same time frame and group of patients. METHODS: The Severity Scoring of Atopic Dermatitis (SCORAD) index was used for assessment of disease severity in 21 adult patients in acute stage of AD and after complete resolution of clinical findings. Serum levels of IL-16, MDC, ECP and sE-selectin were measured at the same time points in 18 patients and compared with healthy nonatopic controls. The correlation between SCORAD and each laboratory parameter was tested for significance and compared. RESULTS: Serum levels of IL-16, MDC, ECP and sE-selectin were significantly higher in patients in acute stage of AD compared with controls and decreased significantly after treatment, in parallel with clinical improvement. All monitored parameters reflected disease severity assessed by the clinical score. We found the highest significance level of correlation with SCORAD for IL-16 (r = 0.68, P =0.0019), followed by ECP (r = 0.65, P = 0.0032) and MDC (r = 0.55, P =0.0326). There was significant correlation between serum levels of IL-16 and MDC (r = 0.53, P = 0.0443) and ECP and sE-selectin (r = 0.48, P = 0.0427). CONCLUSIONS: The study established a significant correlation between serum levels of IL-16 and SCORAD in adult AD patients. We report a significant correlation between IL-16 and MDC, both T helper 2 activation markers. Our data suggested that IL-16 reflects most convincingly disease severity and may be used as a marker in clinical studies preferentially in combination with a clinical activity score. PMID- 16704643 TI - Daily pattern of sweating and response to stress and exercise in patients with palmar hyperhidrosis. AB - BACKGROUND: Focal hyperhidrosis is an embarrassing condition that can have a significant negative impact on patient quality of life. It is characterized by excessive sweating affecting a variety of areas, including the palms. Little is known about the daily pattern of sweating in patients with palmar hyperhidrosis. OBJECTIVES: To compare the variation of sweating in patients with primary palmar hyperhidrosis with healthy individuals during daily activities. METHODS: Twenty patients with primary palmar hyperhidrosis and 20 healthy age- and sex-matched subjects were studied. Each participant self-assessed rates of sweating for 7 days on an hourly basis using a subjective evaluation scale (SES) ranging from 0 to 10. RESULTS: The 3763 assessments showed clear differences between patients and healthy controls (median SES score 5 and 0, respectively; P < 0.0001). Stress and exercise significantly increased SES by scores of 2-5 in both groups, with stress influencing patients more than controls. SES scores in patients varied significantly, from 0 to 2 during mornings and evenings, and between 5 and 6 at mid-day, while scores in control subjects remained consistently close to 0. The pattern of change depended weakly on sex and weekday/weekend distinction. Dynamic responses to stress and exercise in patients had the tendency to return to baseline more slowly than in controls. CONCLUSIONS: Hourly changes in sweating rates can be assessed successfully through self-assessment. Patients with primary palmar focal hyperhidrosis reported significantly increased palmar sweating and daily patterns of sweating showing considerable variation dependent on factors such as time of day and emotional stimuli. PMID- 16704644 TI - Comparative analysis of incidence and clinical features of cutaneous malignant melanoma in Crete (Greece) and southern Germany (central Baden-Wurttemberg). AB - BACKGROUND: Comparative analysis of the incidence rates and epidemiological features of cutaneous malignant melanoma (CMM) between different ethnic groups exposed to varying environmental factors is critical for consideration of the causes of CMM but can also be utilized in a public health approach to control of the disease. OBJECTIVES: To compare incidence rates and clinical features of CMM in a Greek and a central European population (central Baden-Wurttemberg, Germany). METHODS: Incident cases of CMM were traced in all hospitals of the island of Crete for the period 1999-2002. Age-standardized incidence rates per 100 000 inhabitants per year for the European Standard Population were calculated based on the Cretan population statistics. A comparison was performed between the Cretan findings and those of southern Germany as registered by the hospital-based Central Malignant Melanoma Registry, which likewise documents more clinical features than normally recorded by population-based cancer registries. RESULTS: Mean incidence rates in Crete for 1999-2002 were 4.01 per 100 000 inhabitants per year for males and 4.05 for females as compared with 10.6 for males and 11.1 for females in southern Germany. There were striking differences in the clinical characteristics of CMMs, with significantly higher tumour thickness in Crete (median 1.4 mm vs. 0.7 mm in southern Germany). Correspondingly, significantly more nodular melanomas were observed in Crete (29%) as compared with southern Germany (11%). CONCLUSIONS: Incidence of CMM in Crete, with about four cases per 100 000 inhabitants per year, is clearly higher than previously estimated, and there is an urgent necessity for earlier recognition of CMM in Crete. However, the incidence of CMM in southern Germany is much higher. PMID- 16704645 TI - Do patients with skin allergies have higher levels of anxiety than patients with allergic respiratory diseases? Results of a large-scale cross-sectional study in a French population. AB - BACKGROUND: Psychological comorbidity is a known aspect of allergic disease. However, there is recent evidence that a large proportion of allergic patients remains undiagnosed and untreated for psychological disease. In addition, the complexities of the anxiety-allergy relationship, i.e. differences for current and past disease, or differences among allergic disease types, are not well understood. OBJECTIVES: To measure the level of anxiety in a large allergic population in France using a standardized measure, the State/Trait Anxiety Inventory (STAI). METHODS: Allergy patients in France (n = 3939) who visited their allergy specialists participated in the study. The patients completed a questionnaire which was then linked to the questionnaire completed by their physician. Only patients with both subject and physician questionnaire were kept in the analyses. Mean STAI scores for the State (S) and Trait (T) scales were obtained for each allergic disease. ANCOVA models testing group differences on the mean scores, using the categories "current disease", "past disease" and "allergic disease ever", were assessed along with relevant confounders. RESULTS: Allergic rhinitis (AR), asthma and atopic dermatitis (AD) were the most prevalent conditions of the 12 allergic diseases assessed in the study. Women had higher mean STAI S/T scores than men and age was also found to be associated with higher S scores; therefore, both age and gender were included as covariates where relevant. A single ANCOVA model for each STAI scale showed a statistical difference among the various allergic diseases. Using the category "current disease" each allergic disease was assessed separately regarding the presence or absence of that disease. Higher, statistically significant mean STAI scores were found for AD and allergic urticaria on the S scale and for AD on the T scale. Similarly, for the category "allergic disease ever", AD and allergic urticaria reached statistical significance on the S scale, while on the T scale only AD was statistically significant. When patients were assessed for anxiety based on their past disease, asthma, AR and sinusitis were significant on the S scale while asthma and nasal polyps were statistically significant on the T scale. When asthma and AD were tested simultaneously, only the latter was significant. CONCLUSIONS: High mean scores for State and Trait anxiety were mostly associated with AD. PMID- 16704646 TI - A cost-utility analysis of pimecrolimus vs. topical corticosteroids and emollients for the treatment of mild and moderate atopic eczema. AB - BACKGROUND: Conventional treatments for atopic eczema include topical corticosteroids (TCS) and emollients. Pimecrolimus, an immunosuppressant, was licensed in the U.K. in 2003 as an alternative treatment of mild to moderate atopic eczema. OBJECTIVES: To assess the cost-utility of pimecrolimus as a treatment for mild and moderate atopic eczema when compared with conventional treatments which use TCS and emollients. METHODS: A Markov state-transition model was developed to represent the cyclical nature of atopic eczema and provide an economic analysis of cost-utility for treatment alternatives from the perspective of a third party payer (U.K. National Health Service). A range of methods was used to obtain data for transition probabilities, costs and quality of life. These included a systematic review of published effectiveness data, expert opinion, and a utility study conducted by the authors. Separate cohort analyses were modelled to distinguish between children and adult populations and between differing treatment patterns for facial and body eczema. One-way sensitivity analyses and probabilistic sensitivity analysis (using Monte-Carlo simulation) were performed. RESULTS: Baseline cost-utility outputs from the model show that, in all tested scenarios, TCS dominate pimecrolimus (i.e. TCS are both cheaper and more effective). However, the differences in benefits between treatments output by the model are very small. Sensitivity analyses highlight the importance of cost variations in pimecrolimus. Where pimecrolimus is compared with emollient only it is probably cost effective at a willingness-to-pay threshold of 30 000 UK pounds per quality-adjusted life year. CONCLUSIONS: There are likely to be few situations in which the use of pimecrolimus for the treatment of atopic eczema can be justified on economic grounds. Exceptions are likely to be in cases where TCS have been shown to be ineffective, unacceptable due to adverse events, or where a patient is unwilling to accept TCS treatment despite appropriate education and support and emollient alone is the alternative clinical option. PMID- 16704647 TI - 8-Isoprostane is a dose-related biomarker for photo-oxidative ultraviolet (UV) B damage in vivo: a pilot study with personal UV dosimetry. AB - BACKGROUND: Ultraviolet (UV) B irradiation causes visible erythema, which has been linked with DNA damage. However, besides such direct photochemical conformation changes, UVB also induces many indirect photochemical effects in the skin. Lipid peroxidation (LPO) is in this context one of the major pathways by which photo-oxidative stress disturbs cell signalling and promotes photocarcinogenesis and photoageing. So far we lack techniques for visualizing photo-oxidative stress in the skin. Furthermore, LPO has never been linked with individually acquired UVB doses measured by personal dosimetry. OBJECTIVES: Measuring the skin reaction and photo-oxidative damage by LPO in vivo after UVB exposure in a pilot study surveyed by personal dosimetry in order to allow for a correlation analysis of acquired dose, skin reaction and amount of LPO. METHODS: UVB exposure was measured with the opto-electronic X2000-1 (Gigahertz Optik, Puchheim, Germany) and the biological DLR Biofilm (German Aerospace Center DLR, Cologne, Germany) portable dosimeter. The skin reaction following UVB exposure was quantified with a Minolta chromameter (Minolta, Tokyo, Japan) and LPO in vivo was measured by 8-isoprostane generation by means of densitometric analysis of immunohistochemical samples obtained 30 min post-UVB irradiation. RESULTS: Regression analysis revealed significant linear relations between UVB exposures recorded by the dosimeters and colorimetry parameters of the skin reaction. Furthermore, an even better linear relation with higher significance was found between the generation of 8-isoprostane in the skin and the dosimeter readouts. CONCLUSIONS: LPO measured by the generation of 8-isoprostane provides a suitable intrinsic biomarker for photo-oxidative UVB damage in vivo. This study provides a new approach to visualizing photo-oxidative stress in the skin in vivo. Furthermore, future dosimeter readouts can now be set into relation to the expected increase of LPO that can be calculated within the limits of our study. PMID- 16704648 TI - A 52-week randomized safety study of a calcipotriol/betamethasone dipropionate two-compound product (Dovobet/Daivobet/Taclonex) in the treatment of psoriasis vulgaris. AB - BACKGROUND: The calcipotriol/betamethasone dipropionate two-compound product Dovobet/Daivobet/Taclonex(LEO Pharma A/S, Ballerup, Denmark) has been shown to be safe and effective in the treatment of psoriasis for up to 8 weeks. As psoriasis is a chronic disease, long-term treatment may be required, so there is a need to investigate the safety of its use over a longer period of time. OBJECTIVES: To investigate the safety of two treatment regimens involving use of the two compound product over 52 weeks in the treatment of patients with psoriasis. METHODS: Patients (n = 634) were randomized double-blind to treatment with: (i) 52 weeks of the two-compound product (two-compound group); (ii) 52 weeks of alternating 4-week periods of the two-compound product and calcipotriol (alternating group); or (iii) 4 weeks of the two-compound product followed by 48 weeks of calcipotriol (calcipotriol group). Treatments in all groups were used once daily when required. RESULTS: Adverse drug reactions (ADRs) occurred in 45 (21.7%) patients in the two-compound group, 63 (29.6%) in the alternating group and 78 (37.9%) in the calcipotriol group. The odds ratio for an ADR in the two compound group relative to the calcipotriol group was 0.46 (95% confidence interval 0.30-0.70; P < 0.001). ADRs of concern associated with long-term topical corticosteroid use occurred in 10 (4.8%) patients in the two-compound group, six (2.8%) in the alternating group and six (2.9%) in the calcipotriol group; those with the highest incidence were skin atrophy, occurring in four (1.9%), one (0.5%) and two (1.0%) patients, respectively, and folliculitis, in three (1.4%), one (0.5%) and no patients, respectively. CONCLUSIONS: Treatment with the two compound product for up to 52 weeks appears to be safe and well tolerated whether used on its own or alternating every 4 weeks with calcipotriol treatment. PMID- 16704650 TI - Folic acid supplementation during treatment of psoriasis with methotrexate: a randomized, double-blind, placebo-controlled trial. AB - BACKGROUND: The value of folate supplementation in methotrexate (MTX)-treated patients remains controversial. OBJECTIVES: To determine the effect of folic acid (FA) on the efficacy of MTX and the frequency of side-effects associated with MTX therapy. METHODS: A 12-week double-blind clinical trial was conducted in patients with psoriasis stable on their long-term MTX doses but not receiving FA. They were randomized into two arms of either FA 5 mg or placebo daily. MTX doses were not changed throughout the study. Patients were monitored every 3 weeks by the same observer. Assessments included Psoriasis Area and Severity Index (PASI), a visual analogue scale (VAS) of patients' perception of their psoriasis severity and the Dermatology Life Quality Index (DLQI). Adverse events were systematically recorded. Haematological and biochemical monitoring was performed. RESULTS: Twenty-two patients with psoriasis were recruited. Age, sex and weekly MTX doses were similar in both groups. All 22 patients completed the study. The mean PASI in the FA group increased from 6.4 at baseline to 10.8 at 12 weeks. In the placebo group the mean PASI fell from 9.8 at baseline to 9.2 at 12 weeks. The mean change from baseline in the FA group was 4.4 vs. -0.6 in the placebo group (P < 0.05). Similar trends were observed in the changes in VAS and in the DLQI and differences between the groups were significant for both these parameters (P < 0.05). Few adverse effects were reported. CONCLUSIONS: This study suggests that supplementation with FA during long-term MTX treatment reduces the efficacy of MTX in the control of psoriasis. Due to the relatively small sample size and short duration of this study, no conclusions can be drawn regarding the possibility that FA may reduce the side-effects of MTX. PMID- 16704649 TI - Improvement in quality of life with infliximab induction and maintenance therapy in patients with moderate-to-severe psoriasis: a randomized controlled trial. AB - BACKGROUND: Psoriasis has a well-documented, markedly negative effect on patient quality of life. OBJECTIVES: To evaluate the impact of long-term infliximab maintenance therapy on health-related quality of life (HRQoL) in patients with psoriasis. METHODS: The Dermatology Life Quality Index (DLQI) and 36-item Short Form Health Survey (SF-36) were administered as part of the pivotal double-blind, placebo-controlled efficacy and safety EXPRESS study of infliximab in chronic plaque psoriasis. In total, 378 patients with moderate-to-severe psoriasis were enrolled at 32 centres in Europe and Canada. Patients were randomized to receive either placebo or infliximab 5 mg kg(-1) induction at weeks 0, 2 and 6 followed by maintenance every 8 weeks; placebo patients crossed over at week 24 to receive the infliximab induction and maintenance regimen. RESULTS: At week 10, infliximab treated patients had significantly greater improvement in DLQI scores (P < 0.001) and SF-36 physical and mental component summary scores (P < 0.001) than placebo treated patients. Significant improvement (P < 0.001) was also seen in all eight SF-36 subscales, and was greatest for the "Bodily Pain" and "Social Functioning" scales. Significant improvement in HRQoL persisted with maintenance infliximab treatment at week 24 (P < 0.001), with patients achieving a Psoriasis Area and Severity Index score of 0 reporting the greatest benefit. Treatment-related HRQoL improvement remained substantial at week 50. CONCLUSIONS: Infliximab induction and maintenance regimens resulted in rapid, substantial, sustained and clinically meaningful improvement in both dermatology-specific and general quality of life indices in patients with psoriasis, with total clearance resulting in maximum improvement. PMID- 16704651 TI - Bcl-2 and Bax in congenital naevi. AB - BACKGROUND: Melanocytes represent a static component of the epidermis, and the role of apoptosis in basal melanocyte function and melanocytic tumour formation has not been fully elucidated. OBJECTIVES: The aim of this study was to investigate the expression of Bcl-2 anti-apoptotic and Bax apoptotic proteins in congenital naevi in correlation with p-27 protein and Ki-67 proliferative index. METHODS: Our material comprised 30 congenital naevi (eight giant) excised from children aged from 15 days to 14 years old. The immunohistochemical streptavidin biotin method was performed on paraffin sections for the detection of Bcl-2 (cl100/D5), Bax (cl2D2) , Ki-67 (MIB-1) and p-27 (1B4) proteins with monoclonal antibodies. RESULTS: Bcl-2 protein was detected in all cases showing a strong diffuse cytoplasmic expression in >70% of the naevocytes and was preserved in the deeper parts of the naevi. On the other hand, Bax was detected in 13 of the cases, showing a fainter cytoplasmic expression in 40-50% of the naevocytes without any particular topographic distribution. Ki-67 was detected in all cases showing a limited expression in 1-2% of the nuclei mainly in the junctional and upper dermal components. p-27 protein showed a broad diffuse nuclear expression (>70% of the nuclei) in all cases with a particular increase in the deeper parts of the naevi. Bcl-2 expression showed a parallel correlation with p-27 protein. CONCLUSIONS: Broad Bcl-2 expression in congenital naevi suggests that suppression of apoptosis may play an important role in the maintenance of naevocytes despite the low proliferative activity. PMID- 16704652 TI - Interleukin 12 production by monocytes from patients with psoriasis and its inhibition by ciclosporin A. AB - BACKGROUND: Psoriasis is a T-helper (Th)1 cytokine-mediated chronic skin disease and interleukin (IL)-12 has been shown to play a major role in the development of Th1 responses. OBJECTIVES: To elucidate the role of IL-12 in the pathogenesis of psoriasis and to study the effect of ciclosporin A (CsA) on Th1 deviation of this disease. PATIENTS/METHODS: We investigated IL-12 production by stimulated monocytes from patients with psoriasis who were treated with or without CsA. Monocytes were stimulated with interferon-gamma plus lipopolysaccharide (LPS) or Staphylococcus aureus Cowan strain I (SAC). The amount of IL-12 p70 produced by stimulated monocytes was evaluated by enzyme-linked immunosorbent assay. RESULTS: Compared with those from normal controls, LPS- but not SAC-stimulated monocytes from patients with psoriasis produced significantly higher amounts of IL-12. Interestingly, LPS-stimulated monocytes from patients with psoriasis treated with CsA produced significantly decreased amounts of IL-12 compared with those patients not treated with CsA. CONCLUSIONS: Our results suggest that IL-12 production by monocytes may have a critical role in the pathogenesis of psoriasis, and that the therapeutic effect of CsA on psoriasis may be achieved by correcting the deviation of the Th1/Th2 balance. PMID- 16704653 TI - Subcutaneous hyalohyphomycosis due to Cephalotheca foveolata in an immunocompetent host. AB - We report the first case of subcutaneous hyalohyphomycosis caused by the genus Cephalotheca, which has not been reported to cause human infection. A 67-year-old immunocompetent farmer presented with a 10-year history of verrucous erythematous plaques on the right foot dorsum, great toe, heel and sole. Histopathology of the lesions revealed chronic granulomatous inflammation with numerous nonpigmented fungal spores in the dermis. Cultures of biopsy specimens on Sabouraud's dextrose agar for 2 weeks developed into yellowish brown, velvety colonies that subsequently turned black after 8 weeks because of the production of black cleistothecia indicating the teleomorph. On the basis of mycological features, scanning electron microscopic morphology and molecular analysis data, a new species of Cephalotheca was identified and designated Cephalotheca foveolata. Because there was no response to the antifungal agents administered, the patient was successfully treated by surgical excision with skin graft. PMID- 16704654 TI - Early and severe amyloidosis in a patient with concurrent familial Mediterranean fever and pseudoxanthoma elasticum. AB - A young woman patient had early and extensive familial Mediterranean fever (FMF) related amyloidosis and pseudoxanthoma elasticum (PXE). She had the novel G1042S mutation in the ATP-binding cassette subfamily C member 6 (ABCC6) gene, responsible for PXE, and the mutation M694I in MEFV, the FMF gene. Both mutations were homozygous, in agreement with consanguinity in the parents. ABCC6 deficiency may have increased the severity of amyloidosis by increasing the deposition in target tissues of heparan sulphate, which colocalizes spatially and temporally with amyloid proteins, and/or by decreasing the therapeutic activity of colchicine. PMID- 16704655 TI - Epidermal naevus in Proteus syndrome showing loss of heterozygosity for an inherited PTEN mutation. AB - A 3-year-old boy with Proteus syndrome has a novel germline p.Y68D mutation of the PTEN gene inherited from his mother who has Cowden syndrome. In addition, DNA extracted from curettings of his widespread epidermal naevus shows loss of heterozygosity for this mutation. To our knowledge, this has not been described before. PMID- 16704656 TI - The frequency and significance of thiopurine S-methyltransferase gene polymorphisms in azathioprine-treated renal transplant recipients. PMID- 16704657 TI - Invasive aspergillosis due to subungual onychomycosis during treatment for non Hodgkin lymphoma. PMID- 16704658 TI - An audit of the management of cutaneous squamous cell carcinoma according to the multiprofessional guidelines. PMID- 16704659 TI - Comparable risk of herpes simplex virus infection between topical treatments with tacrolimus and corticosteroids in adults with atopic dermatitis. PMID- 16704660 TI - Relative frequency and survival of patients with primary cutaneous lymphomas: data from a single-centre study of 203 patients. PMID- 16704661 TI - Topical mechlorethamine and clobetasol in multifocal primary cutaneous marginal zone-B cell lymphoma. PMID- 16704662 TI - IgE antibodies to penicillin are indicative for but not conclusive proof of penicillin allergy. PMID- 16704663 TI - Successful treatment of Darier disease with topical 5-fluorouracil. PMID- 16704664 TI - True transverse leuconychia with spontaneous resolution during pregnancy. PMID- 16704665 TI - Imatinib-associated lichenoid eruption: acitretin treatment allows maintained antineoplastic effect. PMID- 16704666 TI - Imatinib mesylate causes palmoplantar hyperkeratosis and nail dystrophy in three patients with chronic myeloid leukaemia. PMID- 16704668 TI - Overview and commentary. AB - The California Health Benefits Review Program (CHBRP)--established in 2003 in response to new state legislation aimed at enhancing the evaluation of potential changes in health benefit packages--represents a unique marriage of academic analysis and real-time legislative decision making. CHBRP is based within the University of California (UC) Office of the President and provides analyses to the legislature within a 60-day timeframe on the potential consequences of specific benefit changes under consideration as part of legislative mandates. The consequences examined include current known medical effectiveness of the services for which coverage is to be mandated as well as potential costs and impact on public health considerations associated with the mandate. Teams throughout the University system specialize in analyzing medical effectiveness, costs, and public health impacts and work with a statewide faculty task force and a private actuarial firm to generate literature reviews and analyses in response to legislative requests. These teams work on multiple requests simultaneously, all within the constraints of the legislative calendar. In its first 2 years, CHBRP generated 22 such analyses. PMID- 16704669 TI - Evaluating medical effectiveness for the california health benefits review program. AB - An important aspect of the mandate assessments requested by the California legislature is a review of the scientific and medical literature on the medical effectiveness of the proposed health insurance benefit mandate. Although such a review bears many similarities to effectiveness reviews that might be undertaken for publication as research studies, several important differences arise from the requirements of the California legislation. Our reviews are intended to assist the legislators in deciding whether to support a specific mandate to modify health insurance benefits in a particular way. Thus, our assessments focus on how the scientific literature bears on the proposed mandate, which may involve a complicated chain of potential effects leading from altered coverage to ultimate impact on health. Evidence may be available for only some of the links in the chain. Furthermore, not all the evidence may be directly applicable to the diverse population of California or the subpopulation affected by the mandate. The mandate reviews, including the medical effectiveness analyses, may be used in a potentially contentious decision making setting. The legislative calendar requires that they need to be timely, yet they must be as valid, credible, and based on the best information available as possible. The focus on applicability also implies the need for informed, technical decisions concerning the relevance of the articles for the report, and these decisions need to be made as transparent as possible. These goals and constraints yield an approach that differs somewhat from an investigator-initiated review of the literature. PMID- 16704671 TI - Assessing the public health impact of state health benefit mandates. AB - OBJECTIVE: To document the process used in assessing the public health impact of proposed health insurance benefit mandates in California as part of the California Health Benefits Review Program (CHBRP) to serve as a guide for other states interested in incorporating a public health impact analysis into their state mandated benefit review process. BACKGROUND: As of September 2004, of the 26 states that require reviews of mandated benefit legislation, 25 required an assessment of the cost impact, 12 required an assessment of the medical efficacy, and only 6 had language requiring an assessment of the public health impact. METHODOLOGY: This paper presents the methodology used to calculate the overall public health impact of each mandate. This includes a discussion of data sources, required data elements, and the methods used to quantify the impact of a mandated health insurance benefit on: overall public health, on gender and racial disparities in health outcomes, on premature death, and on the economic loss associated with disease. In addition we identify the limitations of this type of analysis. CONCLUSIONS: The approach that California has adopted to review proposed health benefit mandates represents a leap forward in its consideration of the impact of such mandates on the health of the population. the approach is unique in its specific requirements to address public health impacts as well as the attempt to quantify these impacts by the CHBRP team. The requirement to make available this information to the state government has the potential, ultimately, to increase the availability of health insurance products in California that will maximize public health. PMID- 16704670 TI - The California Cost and Coverage Model: analyses of the financial impacts of benefit mandates for the California legislature. AB - OBJECTIVE: To produce cost estimates of proposed health insurance benefit mandates for the California legislature. DATA SOURCES: The 2001 California Health Interview Survey, 2002 Kaiser Family Foundation/Health Research and Education Trust California Employer Health Benefits Survey, Milliman Health Cost Guidelines, and ad hoc surveys of large health plans were used. STUDY DESIGN: We developed an actuarial model to estimate short-term (1 year) changes in utilization and total health care expenditures, including insurance premiums and out-of-pocket expenditures, if insurance mandates were enacted. This model includes baseline estimates of current coverage and total current expenditures for each proposed mandate. PRINCIPAL FINDINGS: Analysis of seven legislative proposals indicated 1-year increases in total health care expenditures among the insured population in California ranging from 0.006 to 0.200 percent. Even when proposed mandates were expected to reach a large target group, either utilization or cost was sufficiently low to keep total cost increases minimal. CONCLUSIONS: Our ability to develop a California-specific model to estimate the impacts of proposed mandates in a timely fashion provided California legislators during the 2004 legislative session with more-detailed coverage and cost information than is generally available to legislative bodies. PMID- 16704672 TI - An analysis of California Assembly Bill 2185: mandating coverage of pediatric asthma self-management training and education. AB - OBJECTIVE: To summarize for the California Legislature the evidence on the medical effectiveness of pediatric asthma self-management training and education (PASMTE), including the use of peak flow meters, spacers, and nebulizers and the impact that mandated coverage of these services and devices under Assembly Bill (AB) 2185 would have on total health care expenditures, monthly premiums, health services utilization, and the public's health. MEDICAL EFFECTIVENESS FINDINGS: The review of the literature finds that PASMTE is medically effective and has favorable effects on the health of children with symptomatic asthma, as well as reduces asthma-related emergency room visits and hospitalizations. There was inadequate evidence to assess the effectiveness of the three medical devices independently of PASMTE. COST AND UTILIZATION FINDINGS: One-hundred percent of children in health maintenance organization (HMO) plans in California are already covered for PASMTE, with fewer having coverage for the specific medical devices. However, despite full coverage of PASMTE in HMOs, these services are underutilized. We expect that the enactment of AB 2185 would increase utilization of PASMTE among children who are currently covered by 10 percent as a result of increased awareness of current coverage by all HMOs and increased awareness of the importance of these services. We estimate that this increased utilization by children who are already covered may result in a total statewide premiums increase of $170,000 or 0.006 percent, equal to one to two cents per member per month (PMPM). PUBLIC HEALTH FINDINGS: It is estimated that the public health impact of the mandate, as a result of new utilization of PASMTE by 10 percent of children who are already covered, would reduce the number of school days missed because of asthma per year by 158,000; the number of children reporting restricted activity days by 6,020; the number of emergency department visits by 350; and the number of hospitalizations by 1,105. LEGISLATIVE ACTION: AB 2185 passed the legislature after being amended six times. The bill as it was signed into law did not mandate coverage for PASMTE, as all HMOs in California presently reported covering these services. However, the bill retained the mandate for coverage of the three medical devices, as their coverage was not as universal across health plans. PMID- 16704674 TI - State-mandated benefit review laws. AB - OBJECTIVE: To determine which states have laws that require the review of mandated health insurance benefits and describe the various approaches states take in reviewing mandated benefits, as stated in the mandated benefit review (MBR) laws. DATA SOURCES: We queried online databases of the individual state statutes and reviewed the state statutes and state legislative agendas for all 50 states and Washington, DC to identify those states with active MBR laws as of September 2004. STUDY DESIGN: We reviewed the identified MBR laws to catalog their various components. The components chosen for this analysis include: general review strategy, designated reviewers, time frame for conducting reviews, criteria used in the review, requirements to use actuaries, sources of funding, and state data collection systems. Two of the authors independently created analysis categories and coded the MBR laws to document details on the major components of the laws. PRINCIPAL FINDINGS: We identified 26 state MBR laws active as of September 2004. A majority of the MBR laws specified a prospective review approach and only one law used an exclusively retrospective review approach. A substantial amount of variation was found with regards to the designated reviewers, time frames for conducting reviews, and criteria used in the review. Few states specified the use of actuaries, sources of funding, and state data collection systems. CONCLUSIONS: The number of states that have enacted MBR laws has increased substantially in recent years, however, different states have structured the review of mandated benefits differently, according to the values and perceived needs of the state legislatures. It is important that states increasingly consider a broader scope of review criteria so state decision makers can position themselves to mandate only those benefits that add real value to the state's health care system. PMID- 16704673 TI - A comparative analysis of mandated benefit laws, 1949-2002. AB - OBJECTIVE: To understand and compare the trends in mandated benefits laws in the United States. DATA SOURCES/STUDY SETTING: Mandated benefit laws enacted in 50 states and the District of Columbia for the period 1949-2002 were compiled from multiple published compendia. STUDY DESIGN: Laws that require private insurers and health plans to cover particular services, types of diseases, or care by specific providers in 50 states and the District of Columbia are compared for the period 1949-2002. Legislation is compared by year, by average and total frequency, by state, by type (provider, health care service, or preventive), and according to whether it requires coverage or an offer of coverage. DATA COLLECTION/EXTRACTION METHOD: Data from published tables were entered into a spreadsheet and analyzed using statistical software. PRINCIPAL FINDINGS: A total of 1,471 laws mandated coverage for 76 types of providers and services. The most common type of mandated coverage is for specific health care services (670 laws for 34 different services), followed by laws for services offered by specific professionals and other providers (507 mandated benefits laws for 25 types of providers), and coverage for specific preventive services (295 laws for 17 benefits). On average, a mandated benefit law has been adopted or significantly revised by 19 states, and each state has approximately 29 mandates. Only two benefits (minimum maternity stay and breast reconstruction) are mandated in all 51 jurisdictions and these were also federally mandated benefits. The mean number of total mandated benefit laws adopted or significantly revised per year was 17 per year in the 1970s, 36 per year in the 1980s, 59 per year in the 1990s, and 76 per year between 2000 and 2002. Since 1990, mandate adoption increased substantially, with around 55 percent of all mandated benefit laws enacted between 1990 and 2002. CONCLUSIONS: There was a large increase in the number of mandated benefits laws during the managed care "backlash" of the 1990s. Many states now use mandated benefits to prescribe not only what services and benefits would be provided but how, where, and when services will be provided. PMID- 16704675 TI - Health services research as a source of legislative analysis and input: the role of the California Health Benefits Review Program. AB - This article examines the role of the California Health Benefits Review Program (CHBRP) as a source of information in state health policy making. It explains why the California benefits review process relies heavily on university-based researchers and employs a broad set of criteria for review, which set it apart from similar programs in other states. It then analyzes the politics of health insurance mandates and how independent research and analysis might alter the perceived benefits and costs of health insurance mandates and thus political outcomes. It considers how research and analysis is typically used by policy makers, and illustrates how participants inside and outside of state government have used the reports prepared by CHBRP as both guidance in policy design and as political ammunition. Although there is consensus that the review process has reduced the number of mandate bills that are passed out of the legislature, both supporters and opponents favor the new process and generally believe the reports strengthen their case in legislative debates over health insurance mandates. The role of the CHBRP is narrowly defined by statute at the present time, but the program may well face pressure to evolve from its current academic orientation into a more interactive, advisory role for legislators in the future. PMID- 16704676 TI - Comparison of three conservative treatment protocols in carpal tunnel syndrome. AB - The aim of this study was to investigate and compare the therapeutic effect of three different combinations in the conservative treatment of carpal tunnel syndrome (CTS) by means of clinical and electrophysiological studies. The combinations included tendon- and nerve-gliding exercises in combination with splinting, ultrasound treatment in combination with splinting and the combination of ultrasound, splinting, tendon- and nerve-gliding exercises. A total 28 female patients (56 wrists) with clinical and electrophysiologic evidence of bilateral CTS were studied. In all patient groups, the treatment combinations were significantly effective immediately and 8 weeks after the treatment. The results of the long-term patient satisfaction questionnaire revealed that symptomatic improvement is more prominent in the group treated with splinting, exercise and ultrasound therapy combination. Our results suggest that a combination of splinting, exercise and ultrasound therapy is a preferable and an efficacious conservative type of treatment in CTS. PMID- 16704677 TI - The characteristics and outcome of primary vesicoureteric reflux diagnosed in the first year of life. AB - A retrospective trial was performed to study presentation, evaluation, management, complications and outcome of 186 infants with vesicoureteral reflux (VUR). Medical records of 103 male and 83 female infants with mean age at entry 5.97 months were reviewed. Diagnosis was established using radiographic voiding cystourethrogram. At diagnosis, a renal ultrasound and dimercaptosuccinic acid renal scintigraphy were performed in all children. The follow-up included blood pressure measurements, serial urine cultures, haematological and biochemical tests, radionuclide cystography, renal ultrasounds and renal scintigraphy. The majority of infants with reflux, 176/186, presented with one or more episodes of urinary tract infections. In 113 children, reflux resolved spontaneously, 27 underwent surgical or endoscopic correction and 46 are being followed-up to date. Spontaneous resolution after prophylaxis was more frequent in boys (p < 0.0001), in children with grade I or II (p < 0.0001) and unilateral reflux at diagnosis (p = 0.0215). No significant difference could be established with respect to the presence of scars (p = 0.1680) and the number of breakthrough urinary tract infections (p = 0.1078). The data of the present study indicate that spontaneous resolution rate is high in infants, and therefore, early antireflux PMID- 16704678 TI - Anticardiolipin antibodies and vascular access thrombosis in Taiwanese haemodialysis patients with chronic hepatitis C: a retrospective study. AB - Hepatitis C virus causes various extrahepatic immunologic abnormalities. Vascular access thrombosis (VAT) is a major cause of morbidity in chronic haemodialysis (HD) patients. Immunoglobulin-G anticardiolipin antibody (IgG-ACA) is strongly associated with venous and arterial thrombosis in patients with normal renal function. Previous investigations have reported the association of raised IgG-ACA titre recurrent with VAT in HD patient, and also few equivalent studies were reported the same in Taiwan. This study attempted to determine whether raised IgG ACA titres are associated with increased risk of recurrent VAT in HD patients with chronic hepatitis C. This study enrolled 98 chronic hepatitis C patients undergoing HD. IgG-ACA titre and hepatitis C marker were measured for all subjects. Raised IgG-ACA titres were present in 29.6% (29/98) of patients. In both groups (raised and normal IgG-ACA), the type of shunt did not differ (p = 0.416). There was strong association between raised IgG-ACA titre and recurrent VAT (p = 0.0004). In predicting for more or one episodes of VAT using multiple logistic regression, synthetic graft (p < 0.0001), raised IgG-ACA titre (p = 0.039), presence of hepatitis B (p = 0.004) and haemodialysis duration (p = 0.039) were significant factors. The prevalence of raised IgG-ACA titres was 39.6% among chronic hepatitis C with HD patients. There was strong association between raised IgG-ACA titre and recurrent VAT, and this finding may be the consequence of pathogenetic role of raised IgG-ACA titres on the development of VAT status in HD patients with chronic hepatitis C. PMID- 16704679 TI - Lactococcus garvieae infections in humans: possible association with aquaculture outbreaks. AB - Lactococcus garvieae is an important pathogen in aquaculture, outbreaks of which significantly affect production. It is a rare pathogen with a low virulence in human infection. The relation between the aquaculture outbreak and the human infection has not been clarified. Prospective and retrospective epidemiologic surveillance of the four patients with L. garvieae infection between 2000 and 2003 and their relations to the aquaculture outbreaks of L. garvieae were conducted. All the four patients with L. garvieae infection were associated with gastrointestinal disorders. Three of the four patients gave a history of consuming raw fish and in three of the four patients, the infection occurred in summer between June and August while there is a decrease of fisheries production and an increase in L. garvieae infection in aquaculture farms. There was a 100% identity of 16S rDNA sequence of L. garvieae isolates from patient 1 and from the squid muscle obtained from the restaurant where patient 1 consumed the raw fish. Sporadic occurrence of L. garvieae infection in human appears to correlate with the seasonal aquaculture outbreaks of L. garvieae infection. The presence of gastro-intestinal disorder may facilitate L. garvieae infection. PMID- 16704680 TI - Serum levels of IGF-I and IGFBP-III and their relation with carcinoembryonic antigen and carbohydrate antigen 19-9 in cases of esophageal cancer. AB - Tumour markers are used for diagnosis, staging, evaluation of response to treatment, prognosis and detection of recurrences in clinical oncology. In this study, we aim to investigate the levels of insulin-like growth factor (IGF)-I and IGF-binding protein (IGFBP)-III in cases with oesophageal carcinoma. We investigated their possible use as tumour markers and their relation to other tumour markers. Forty patients who were diagnosed as having oesophageal carcinoma by histopathological evaluation of endoscopic biopsies between January 2003 and July 2004 and 40 healthy people as the control group were included in the study. The serum levels of tumour markers including IGF-I, IGFBP-III, carcinoembryonic antigen (CEA) and carbohydrate antigen (CA) 19-9 were measured in both groups. Data were compared statistically, and the importance of IGF-I and IGFBP-III levels were investigated in cases with oesophageal carcinoma. IGF-I levels were significantly higher in patients with oesophageal carcinoma when compared with the control group (p < 0.05), whereas IGFBP-III levels were significantly lower (p < 0.05). The increase in CEA levels was not statistically significant when compared with the control group. The increase in CA 19-9 levels was statistically significant when compared with the control group (p < 0.05). No correlation was detected between levels of IGF-I and IGFBP-III and levels of CEA and CA 19-9. We suggest that the serum IGF-I level may be used as a tumour marker in oesophageal carcinoma. A low level of serum IGFBP-III is also significant in cases with oesophageal carcinoma. We believe that drugs which inhibit IGF-I function or which stimulate the function of IGFBP-III may open new horizons in extra-surgical modalities for the treatment of oesophageal cancer. PMID- 16704681 TI - Pneumocephalus: a case report and review of article. AB - A 50-year-old woman with a history of ventriculopleural shunt placement, presented with nausea, vomiting, fever, respiratory distress and altered mental status. Computerised tomography scan of head revealed presence of air in the ventricles. Source of air was most probably an episode of resolved spontaneous pneumothorax, as no pneumothorax was present in the admission chest X-ray. Shunt revision was planned emergently, and broad-spectrum antibiotics were started. The patient recovered and was discharged in her usual state of health. PMID- 16704682 TI - Hyperhomocysteinaemia and vascular access thrombosis among chronic hemodialysis patients in Taiwan: a retrospective study. AB - Vascular access thrombosis (VAT) is an important cause of morbidity for chronic haemodialysis (HD) patients. Some risk factors for VAT have been well-defined for chronic HD patients from western countries. However, only a few such factors have been confirmed for Taiwanese patients. This study attempted to determine the association between hyperhomocysteinaemia and the incidence of VAT for chronic HD patients in Taiwan. We retrospectively enrolled a total of 196 patients into this study during 2003. The patients were separated into VAT (n = 142) and control (n = 54) group. The participants of the VAT group were identified as those having one or more VAT, and the participants of the control group were those with no VAT in the past. The mean follow-up period was 48 months. The mean serum homocysteine levels were 29.5 +/- 9.6 and 29.1 +/- 9.5 micromol/l for the VAT (n = 142) and the control (n = 54) group, respectively. There was no significant difference in the level of homocysteine between the VAT and the control group (p = 0.70). Female chronic HD patients had significantly greater mean total homocysteine levels than male (30.89 micromol/l, 95% CI 28.84-32.94 vs. 28.06 micromol/l, 95% CI 26.32-29.82, respectively, p = 0.038). That synthetic graft was a significant risk factor for VAT was determined using multivariate logistic regression analysis. There was no association between serum total homocysteine levels and the incidence of VAT in chronic HD patients in Taiwan. PMID- 16704683 TI - Working in developing countries: the challenges and the satisfaction. PMID- 16704684 TI - Ocular angiogenesis: the role of growth factors. AB - Ocular angiogenesis, the formation of new vessels from the existing vascular tree, is a major cause of severe vision loss. It can affect different structures in the eye, including the retina, choroid and cornea. During the last decade our knowledge in the mechanisms underlying ocular angiogenesis has increased dramatically. We have witnessed the identification of key molecules. Many are classified as growth factors due to their biological properties, regulating angiogenesis. This knowledge has propelled the development of a new group of therapeutic tools, the antiangiogenic agents. This review gives an update on the role of growth factors in ocular angiogenesis from both a basic and a clinical perspective. PMID- 16704685 TI - Intravascular oxygen saturation in retinal vessels in normal subjects and open angle glaucoma subjects. AB - PURPOSE: To measure the oxygen saturation (SO(2)) in retinal arterioles and venules in patients with glaucomatous optic neuropathy. METHODS: We examined SO(2) in retinal arterioles and venules simultaneously by imaging spectrometry. Oxygen saturation was evaluated according to the difference of the extinction spectra of haemoglobin and oxyhaemoglobin. The arterio-venous difference (avD) was calculated by (SO(2art) - SO(2ven)). The optic nerve head topography was estimated by Heidelberg retinal tomography and the visual field using the Octopus G1. We examined one eye in each of 58 healthy persons (mean age 58.6 +/- 10.7 years; mean rim area 1.52 +/- 0.33 mm(2); mean defect 0.65 +/- 1.31 dB; mean intraocular pressure [IOP] 18.5 +/- 2.7 mmHg), 49 patients with normal-tension primary open-angle glaucoma (NTG) (mean age 63.0 +/- 8.5 years; mean rim area 0.89 +/- 0.34 mm(2); mean defect 5.4 +/- 4.1 dB; mean IOP 19.2 +/- 2.9 mmHg), and 45 patients with high-tension primary open-angle glaucoma (POAG) (mean age 62.6 +/- 10.3 years; mean rim area 0.97 +/- 0.47 mm(2); mean defect 7.1 +/- 6.4 dB; mean IOP 31.6 +/- 10.8 mmHg). RESULTS: The intraclass correlation coefficients of the SO(2) measurement were 0.82 (arteriole) and 0.59 (venule). In normal eyes, the SO(2art), SO(2ven) and avD were 92.3 +/- 3.4%, 55.7 +/- 6.8% and 36.6 +/- 7.0%, respectively. Equivalent data were 89.7 +/- 5.4%, 56.0 +/- 8.3% and 33.7 +/ 10.6%, respectively, in NTG eyes and 91.4 +/- 4.0%, 58.3 +/- 10.5% and 33.1 +/- 11.5%, respectively, in POAG eyes. Over all examined eyes, the arteriolar SO(2) and the retinal arterio-venous difference correlated significantly with the rim area. CONCLUSION: Eyes with NTG showed significantly decreased arteriolar SO(2). These changes were not seen in POAG patients. PMID- 16704686 TI - A 3-dimensional evaluation of the macular region: comparing digitized and film based media with a clinical evaluation. AB - PURPOSE: To compare the results of stereoscopic macular evaluations using three different types of stereoscopic photographic equipment with those of a clinical examination when establishing the presence of possible macular oedema. METHODS: The study involved a clinical evaluation of the macular region in 57 eyes of 30 diabetes subjects with possible macular oedema. Clinical examinations were followed by photographic stereoscopic evaluations of the macular region using two digital systems (low and high resolution) and one film-based system. All the photographic recordings were classified by two consultants, who were trained in making macular gradations and blinded to the results of previous assessments. RESULTS: The low-resolution digital equipment was found to have relatively low optical resolution and found a significantly lower number of eyes with macular oedema compared with the clinical examination. The high-resolution digital equipment and the film-based equipment were found to have relatively high optical resolution. The number of eyes with macular oedema found by the high-resolution digital and film-based evaluations was comparable with the number found by the clinical examination. Low intra- and interobserver agreement was observed. CONCLUSION: All three photographic systems have disadvantages that make them suboptimal as tools for evaluating the presence of macular oedema. The film-based system seems the most reliable, while low-resolution digital equipment seems least useful. The development of new and objective measurements giving a 3 dimensional presentation of the macular region might be advantageous. PMID- 16704687 TI - The presence of an operculum verified by optical coherence tomography and other prognostic factors in macular hole surgery. AB - PURPOSE: To evaluate the prognostic influence of the presence of an operculum in macular hole and to correlate the prognosis for postoperative visual acuity (VA) with the size of the macular hole, its duration and preoperative VA. METHODS: We carried out a prospective study of 61 eyes of 59 patients with full thickness macular hole. All patients were examined with optical coherence tomography (OCT) in order to ascertain the presence of an operculum. The VA was measured with the ETDRS chart. A standard pars plana vitrectomy with inner limiting membrane peeling was performed in all cases and 20% C2F6 was used as tamponade. RESULTS: Anatomical closure was obtained after one surgical procedure in 59/61 (97%) patients. Linear regression analysis showed only baseline VA to be predictive of visual outcome (p < 0.001). The presence of an operculum preoperatively did not correlate with VA either pre- or postoperatively, nor with the anatomical closure rate. CONCLUSION: The finding that postoperative VA in macular hole correlates with preoperative VA seems to allow for good prediction of postoperative VA in macular hole surgery. However, the presence of an operculum is of no prognostic significance. PMID- 16704688 TI - Angiotensin-converting enzyme insertion/deletion polymorphism and retinal artery occlusion. AB - PURPOSE: An insertion/deletion (I/D) polymorphism of the gene for angiotensin converting enzyme (ACE) is associated with higher ACE plasma levels and activity. This enzyme is known to play an important role in blood pressure regulation and the ACE I/D gene polymorphism has been suggested as a risk factor for atherosclerotic vascular diseases. The purpose of the present study was to investigate a hypothesized association between the ACE I/D polymorphism and retinal artery occlusion (RAO). METHODS: A total of 159 patients with RAO and 304 control subjects were enrolled in the present retrospective case-control study. ACE I/D genotypes were determined by polymerase chain reaction. RESULTS: Allelic frequencies and genotype distribution of the ACE I/D polymorphism did not significantly differ between patients and control subjects (ACE DD 25.8% versus 28.0%; p = 0.36). A logistic regression analysis predicted the presence of RAO by arterial hypertension and current smoking status, but not by ACE I/D genotypes. CONCLUSION: Our data suggest that the ACE I/D polymorphism is not a major risk factor for RAO. PMID- 16704689 TI - Cytokine gene polymorphisms in retinal detachment patients with and without proliferative vitreoretinopathy: a preliminary study. AB - PURPOSE: Cytokines and other growth factors such as interleukins play an important role in the pathogenesis of proliferative vitreoretinopathy (PVR). Interindividual variations in cytokine production seem to correlate with some cytokine gene polymorphisms. The purpose of this study was to analyse the distribution of these cytokine gene variants in patients with rhegmatogenous retinal detachment (RD) with and without PVR. METHODS: Single nucleotide polymorphisms were analysed for five cytokines: tumour necrosis factor-alpha (TNF alpha), transforming growth factor-beta1 (TGF-beta1), interferon-gamma (IFN gamma), interleukin-6 (IL-6) and interleukin-10 (IL-10). Patients were divided into two surgically treated groups of RD patients: group RD had 27 patients with RD, and group PVR had 31 patients with RD complicated by PVR. A control group was composed of 46 ethnically matched healthy individuals. RESULTS: The genotype distribution of the TGF-beta1 codon 10 polymorphism differed between PVR and RD patients (p = 0.018) and between PVR patients and controls in codon 25 (p = 0.011). There was a higher frequency of TGF-beta1 codon 10 allele T in PVR patients compared with RD patients (p = 0.023). No statistically significant differences between groups were observed for the other polymorphisms examined. CONCLUSION: An association between the TGF-beta1 genetic profile and the development of PVR was detected in this study. Further studies are necessary to confirm this finding and to establish its clinical relevance. PMID- 16704690 TI - Macular oedema in central retinal vein occlusion treated with intravitreal triamcinolone. AB - PURPOSE: To investigate the efficacy of intravitreal triamcinolone as treatment for macular oedema in central retinal vein occlusion (CRVO). METHODS: We conducted a retrospective comparative case series of nine patients with macular oedema associated with CRVO (six non-ischaemic and three ischaemic) treated with an intravitreal injection of 4 mg triamcinolone acetonide, compared with 10 control (observation) patients (six non-ischaemic and four ischaemic). Examination included visual acuity (VA) tests and complete ophthalmic examinations at baseline, 1, 2 and 6 months postoperatively. RESULTS: The mean baseline VA was 20/161 for CRVO treatment group patients and 20/75 for observation group patients (p = 0.15). No significant difference in VA between CRVO treatment group patients (20/99) and controls (20/282) was observed at the final 6-month visit (p = 0.33). Subgroup analysis of the non-ischaemic CRVO treatment patients compared with the non-ischaemic controls also showed no significant difference at the 6-month visit (20/59 and 20/100, respectively; p = 0.20). At 6 months, five of the six non-ischaemic treated patients had VA >or= 20/100, compared with five of the six non-ischaemic control patients. All patients tolerated the procedure well, but there was a significant increase in intraocular pressure by the 2-month visit (p = 0.015). CONCLUSIONS: Intravitreal injection of triamcinolone may not be effective for treatment of macular oedema in all CRVO patients or all non-ischaemic CRVO patients. A trend towards VA improvement was noted but was not statistically significant. Although our treatment was not hindered by severe complications, there was a significant increase in IOP in the 2 months following treatment. PMID- 16704691 TI - Effect of topical steroids on corneal epithelial healing after vitreoretinal surgery. AB - PURPOSE: Topical steroid use is usually avoided in cases of corneal epithelial defect. We evaluated the effect of topical steroid treatment on corneal epithelial healing after epithelial debridement in vitreoretinal surgery. METHODS: Our study population included 85 eyes undergoing vitreoretinal surgeries in our clinic. We prospectively compared the duration of corneal epithelial wound healing in 43 eyes in which topical dexamethasone was used with that in 42 eyes in which topical dexamethasone was not used in the early postoperative period after epithelial debridement. Factors that may retard corneal epithelial healing, including pre- and intraoperative topical solutions, median operative time, the presence of diabetes mellitus, prior ocular surgeries, pseudophakia, aphakia and the presence of intraocular gas or silicone oil in aphakic patients, were not significantly different between the two groups. RESULTS: The mean corneal epithelial defect closure time was 59.7 +/- 2.6 hours (mean +/- SEM) in the group receiving topical steroid treatment, and 61.9 +/- 2.6 hours in the group that did not receive steroids. CONCLUSION: Topical dexamethasone administered five times/day did not significantly retard corneal epithelial healing in subjects undergoing vitreoretinal surgery with postoperative topical steroid treatment, compared with subjects who did not receive steroid treatment. PMID- 16704692 TI - Choroidal neovascular membrane in a series of cases of malignant melanoma of the choroid. AB - PURPOSE: To examine a series of choroidal melanoma specimens to determine the frequency of overlying choroidal neovascularization (CNV) and to ascertain whether CNV over choroidal malignant melanoma is associated with any particular histological tumour characteristics. METHODS: We carried out a retrospective histological analysis of globes containing choroidal melanomas for evidence of choroidal neovascular membranes. RESULTS: Microscopic evidence of choroidal neovascular membranes was evident in 6% of cases. Choroidal neovascularization was not associated with any particular histological tumour characteristic. CONCLUSION: Choroidal neovascularization over choroidal malignant melanoma is not an infrequent occurrence and possibly appears as frequently as CNV over choroidal naevi. The presence of a choroidal neovascular membrane over a pigmented fundal lesion should not be taken as reassurance that the lesion is benign. PMID- 16704693 TI - Transpupillary thermotherapy for the treatment of occult CNV in age-related macular degeneration: a prospective randomized controlled pilot study. AB - PURPOSE: To evaluate whether transpupillary thermotherapy (TTT) reduces the risk of moderate visual loss in patients with occult choroidal neovascularization (CNV) secondary to age-related macular degeneration (AMD). METHODS: A group of 25 patients were recruited and randomized into TTT or placebo groups. Patients were included if they had a subfoveal purely or predominantly (> 50%) occult CNV secondary to AMD with best corrected visual acuity (BCVA) of 6/60 or better and the lesion was not larger than 4.5 mm. Treatment was carried out using an 810-nm Oculight diode laser with a fixed spot size covering the whole lesion according to the standard protocol. The same procedure was used for the control group, except that the power was set at zero. The patients were followed up at 6 weeks, 3 months and then every 6 months for up to 2 years. A maximum of three treatments were administered in both groups if there was evidence of persistent leakage from CNV. RESULTS: At the 12-month follow-up, there was no significant difference in the mean values for BCVA distance and near or contrast sensitivity between the treatment and control groups. The Mann-Whitney test was used to assess the differences in BCVA and contrast sensitivity between the groups, both at baseline and at the 12-month follow-up. No statistically significant difference was found; both groups lost on average two lines of BCVA. CONCLUSION: Transpupillary thermotherapy appeared to have been of no benefit in preventing further visual loss in patients with occult CNV in this pilot study. PMID- 16704694 TI - Chlamydia pneumoniae is not detectable in subretinal neovascular membranes in the exudative stage of age-related macular degeneration. AB - PURPOSE: Age-related macular degeneration (AMD) is the most frequent cause of severe visual impairment in western countries, but its aetiology remains unclear. A growing body of evidence suggests that inflammation contributes to the pathogenesis of AMD, similarly to that shown for atherosclerosis. In view of a number of shared risk factors between the two entities and the hypothesized link between Chlamydia pneumoniae infection and atherosclerosis, we investigated whether C. pneumoniae might be involved in exudative AMD. METHODS: To examine whether C. pneumoniae contributes to the development of subretinal neovascular (SRNV) membranes in AMD, 13 consecutive SRNV membranes surgically excised from patients with exudative AMD were collected and assayed for the presence of C. pneumoniae or other bacterial pathogens by means of polymerase chain reaction (PCR). RESULTS: The age of patients ranged from 68 to 85 years (median 73.5 years). In all 13 SRNV membranes, no DNA of either C. pneumoniae or other pathogens was found by PCR. CONCLUSIONS: These findings indicate that C. pneumoniae is not associated with the development of SRNV membranes in exudative AMD. PMID- 16704695 TI - Scotoma mapping by semi-automated kinetic perimetry: the effects of stimulus properties and the speed of subjects' responses. AB - PURPOSE: The study aimed firstly to determine the effects of stimulus variables on the detection of a scotoma border and, secondly, to study the reproducibility of the method during semi-automated kinetic perimetry. METHODS: The size of the physiological blind spot in 18 young normal subjects was measured with a video campimetric device, the Tubingen computer campimeter (TCC). Kinetic stimuli of two different sizes and at four different levels of luminance were presented. Examinations were repeated within 2 weeks. Measurements were corrected for individual response times and the area of the blind spot was computed. The effects of stimulus strength and size and the repeatability of blind spot measurements were evaluated by an analysis of variance. RESULTS: The physiological blind spot showed significant inter- and intraindividual variations in size (least square means ranging from 17 to 49 square degrees), with a standard deviation of 6.8 square degrees. The measured size of the blind spot increased as a function of decreasing stimulus value, by reducing either the relative brightness or the size of stimuli. Use of a correction for each subject's speed of responses nearly halved the level of random variance. The temporal sequence of measurements (the order in which they were performed) had no apparent effect on the calculated values of blind spot size. CONCLUSIONS: Semi automated kinetic perimetry can determine the size of the physiological blind spot with good repeatability in young, normal subjects. Determination of each individual's speed of response and inclusion of this variable in the calculations reduced variance of the measure significantly. This study confirmed the presence of considerable interindividual differences in the size of the physiological blind spot. PMID- 16704696 TI - Sensitivity and specificity of frequency-doubling technology, tendency-oriented perimetry, SITA Standard and SITA Fast perimetry in perimetrically inexperienced individuals. AB - PURPOSE: To evaluate the sensitivity and specificity of the screening modes of frequency-doubling technology (FDT), tendency-oriented perimetry (TOP), SITA Standard (SS) and SITA Fast (SF) in perimetrically inexperienced individuals. METHODS: One eye of 64 glaucoma patients and 53 normal subjects who had never undergone automated perimetry were tested with programs C-20-5 (FDT), G1 (TOP) and 24-2 (SS and SF). The gold standard for glaucoma was the presence of a typical glaucomatous optic disc appearance on stereoscopic examination (judged by a glaucoma expert), and intraocular pressure (IOP) > 21 mmHg. The test order among strategies was randomized for each subject. To define an abnormal visual field, we applied three criteria for SS and SF and two criteria for TOP and FDT, all of which have been previously described in the literature. Sensitivities and specificities among the different criteria were compared using the Cochran test. RESULTS: Frequency-doubling technology showed the shortest mean test duration, followed by TOP, SF and SS (p < 0.05). Sensitivity ranges were 87.5-89.1% for SS, 92.2-93.8% for SF, 87.5-89.1% for TOP, and 82.8-85.9% for FDT (p = 0.34). Specificity ranges were 73.6-83% for FDT, 56.6-62.3% for TOP, 60.4-69.8% for SF and 66.0-71.7% for SS. The specificity obtained with criterion 2 for FDT (based on the presence of two or more abnormal locations regardless of the severity of abnormal points) was higher than those measured with the other strategies (p < 0.01). CONCLUSION: When testing individuals with no perimetric experience, moderate sensitivities and specificities should be expected, regardless of the strategy chosen. PMID- 16704697 TI - Visual field testing with the new Humphrey Matrix: a comparison between the FDT N 30 and Matrix N-30-F tests. AB - PURPOSE: To determine if the N-30-F test with the new Humphrey Matrix instrument is comparable to the traditional frequency-doubling technology (FDT) N-30 test. METHODS: A sample of 47 eyes from 47 patients (five normal subjects, 27 with ocular hypertension and 15 with glaucoma) underwent visual field (VF) testing with standard automated perimetry, and with FDT, using the FDT N-30 program, and the Matrix N-30-F program (using the same 19 tested points, full-threshold strategy, but with an enhanced algorithm to reduce testing time). The plots, mean deviation (MD), pattern standard deviation (PSD), defect locations and test time were compared. RESULTS: The agreement between the two tests was shown to be good, except for subjects with large VF defects, in which greater PSD values were observed with the FDT test, whereas the Matrix test gave a higher MD and number of abnormal test locations in the total deviation plot. A lack of agreement was also found for the number of abnormal test locations on the pattern deviation plot at any VF loss severity level. The test time for the Matrix test was significantly shorter. CONCLUSION: The FDT N-30 and Matrix N-30-F tests proved to be comparable in cases of early to moderate VF defects. PMID- 16704698 TI - Visual field defects in prematurely born patients with white matter damage of immaturity: a multiple-case study. AB - PURPOSE: White matter damage of immaturity may affect visual, motor and cognitive functions. This multiple-case study presents standardized perimetry results in six teenagers and young adults born prematurely with visual dysfunction due to white matter damage of immaturity of pre- or perinatal origin. METHODS: Six subjects, aged 13-25 years, born at a gestational age of 28-34 weeks, with white matter damage of immaturity documented by MRI, and optic disc appearances documented by fundus photography, were examined with manual and computerized quantitative perimetry. RESULTS: All subjects had subnormal visual field (VF) function, although the depth and extension of the VF defects differed between subjects. The inferior VF function was more deviant than the superior in all cases. The concordance between the VF defects detected with the different techniques was good, although the static computerized techniques revealed slightly more abnormality. CONCLUSION: White matter damage of immaturity may affect the VF. The lower VF is often more affected than the upper. The abnormalities can be demonstrated by both manual and computerized perimetry. PMID- 16704699 TI - Cost-effectiveness analysis in glaucoma: what drives utility? Results from a pilot study in Sweden. AB - PURPOSE: To investigate the effect of different levels of visual field defect in glaucoma on utilities and to test if utilities could be assessed using a general questionnaire such as the EQ-5D. METHODS: A cross-sectional study in 199 patients with ocular hypertension or open-angle glaucoma grouped into 5 severity stages according to visual field defects was performed in 4 specialized ophthalmic centres. Descriptive analysis was performed for the sample and by stage, and the effect of vision loss on utility was investigated with multiple step-wise regression analysis. RESULTS: The mean age of the sample was 70 and the mean MD in the worse eye was -13.1 dB (SD 10.2). Visual acuity (VA) was 0.63 and 0.87 in the worse and better eye, respectively, and the mean utility was 0.80 (SD 0.23). Utility decreased with increasing glaucomatous damage, ranging from 0.84 for mild disease to 0.72 for severe damage (MD -2.5 to -28.1), but the difference between the groups was not statistically significant when controlling for co-morbidity, except for the most severe stage (p < 0.01). In multiple regression analysis, visual field in the better eye was significantly correlated with utility, and there was an indirect correlation between visual field in the worse eye and utility: the effect of total VA on utility was significant, and MD in the worse eye was correlated with total VA. CONCLUSIONS: Utility is strongly correlated with overall vision. Our results suggest a relationship between glaucomatous damage and utility, and patients with severe damage have a significantly lower utility. However, this should be further investigated in larger samples that include more patients with moderate-severe bilateral damage. PMID- 16704700 TI - Fibrin glue for preventing immediate postoperative hypotony following glaucoma drainage implant surgery. AB - PURPOSE: To prevent a leak of aqueous around the tube in the immediate postoperative period after glaucoma drainage implant (GDI) surgery. METHODS: A total of 42 eyes of 34 consecutive patients with refractory glaucoma requiring single-plate Molteno implantation were retrospectively reviewed. Peritubular filtration was checked intraoperatively in all filtered eyes. Fibrin glue was used over the scleral flap intraoperatively in every eye with peritubular leakage. All Molteno tubes were completely closed with an absorbable ligature. RESULTS: Peritubular filtration was detected in 11 eyes of 11 patients. All 11 eyes maintained intraocular pressure (IOP) >or= 16 mmHg in the immediate postoperative phase. The mean IOP on the first postoperative day was 30.5 +/- 10 mmHg. After an average follow-up of 6 months, the mean IOP in these 11 eyes was 19.1 +/- 6 mmHg. No complications or Seidel-positive aqueous leak were observed during the follow-up period. CONCLUSIONS: Results suggest that intraoperative use of fibrin glue is a viable option for reducing peritubular filtration and preventing immediate postoperative hypotony after GDI surgery. PMID- 16704701 TI - The effect of ageing on retinal nerve fibre layer thickness: an evaluation by scanning laser polarimetry with variable corneal compensation. AB - PURPOSE: To evaluate whether a significant age-related thinning of the retinal nerve fibre layer (RNFL) is measurable by means of scanning laser polarimetry with variable corneal compensation (GDx-VCC) in a sample of healthy eyes. METHODS: A sample of 324 eyes of 324 healthy subjects (mean age 56 +/- 14 years, range 21-85 years) underwent a complete ophthalmic evaluation, standard automated perimetry and RNFL scan with the GDx-VCC. Temporal-superior-nasal-inferior temporal (TSNIT), superior and inferior average (SA and IA) values provided on the printout were collected and their mean value averaged from all eyes and also after separating the eyes by gender and by age decades. The values of the three parameters were plotted against age and linear regression was calculated. RESULTS: Ageing is associated with a significant RNFL thinning (0.08 micro, 0.16 micro and 0.12 micro per year for TSNIT, SA and IA, respectively; p < 0.001). Analysis showed a similar association with age decade (p < 0.001 on anova). For the TSNIT average, a 9.5% thinning from baseline values was estimated for a 65 year lifespan. Corresponding values for SA and IA were 16.2% and 11.7%, respectively. CONCLUSIONS: Analysis by GDx-VCC confirmed previous reports about significant age-related RNFL thinning. However, a lower rate per year was found, probably because GDx-VCC measurements are much more reliable than those obtained with the previous generation of polarimeters. PMID- 16704702 TI - Implantation of single-piece foldable acrylic IOLs in small children in the Ukraine. AB - PURPOSE: To evaluate the efficacy and outcome of implantation of a single-piece foldable acrylic intraocular lens (IOL) in children younger than 2 years in the Ukraine. METHODS: A total of 31 children from all over the Ukraine with a median age of 11 months (range 2-28 months) underwent cataract surgery with anterior and posterior capsulorhexis, implantation of a single-piece AcrySof SA30AL IOL and anterior dry vitrectomy. Of these, 22 patients attended the last follow-up. Ten patients had follow-up periods of 12 months; 12 had follow-up periods of 18 months. At the postoperative visits, we investigated visual acuity (VA), whether or not the subject had strabismus, lens epithelial cell migration and signs of secondary glaucoma. RESULTS: The single-piece IOL remained clinically centred in all eyes. No secondary glaucoma was detected. Eleven patients needed surgery for after-cataract. All but two patients achieved better VA after surgery. Many patients suffered from deprivation amblyopia as quite a few cataracts were probably congenital; however, the improvement in VA shows that cataract surgery was worth performing despite the late intervention. CONCLUSION: The foldable single-piece acrylic IOL seems to be well tolerated by children aged 0-2 years. No serious complications were seen and no patient developed secondary glaucoma within the follow-up time. However, lens epithelial cells are very lively in this age group. PMID- 16704703 TI - The use of peribulbar anaesthesia in paediatric cataract surgery (age 7-15 years) in a mobile eye camp in China. AB - AIM: To evaluate the feasibility and acceptability of the use of peribulbar anaesthesia (PA) in paediatric cataract surgery in rural areas in China, where there are limitations in expertise and equipment for general anaesthesia (GA). METHODS: We prospectively evaluated the feasibility and acceptability of carrying out paediatric cataract surgery under PA in children aged 7-15 years. Informed consent was obtained from the subjects and their parents. Children were assessed for their suitability for PA. Peribulbar anaesthesia was given as a peribulbar block using a 2% lidocaine, 0.5% bupivacaine-hyaluronidase mixture administered before lens aspiration with intraocular lens implantation. The acceptability of the PA was evaluated by questionnaire. RESULTS: A total of 19 patients were recruited. Their mean age was 12 +/- 2 years (range 7-15 years). None required conversion to GA. All subjects regarded PA as either totally acceptable (63.2%) or acceptable (36.8%). The mean pain scores (from 0 to 100) during the injection and surgery were 28 +/- 26 and 6 +/- 8, respectively. Seventeen patients (89.5%) said they would prefer PA if choices in anaesthesia were offered again. CONCLUSIONS: Peribulbar anaesthesia can be considered as a viable option in selected children undergoing cataract surgery when facilities for safe and optimal general anaesthesia are unavailable. PMID- 16704704 TI - Tilt and decentration of the intraocular lens following combined vitrectomy and pars plana lensectomy. AB - PURPOSE: To investigate intraocular lens (IOL) tilt and decentration following combined vitrectomy and pars plana lensectomy (PPL) with IOL implantation in patients with proliferative diabetic retinopathy. METHODS: We followed 25 patients with proliferative diabetic retinopathy who underwent PPL and IOL (MA60BM) implantation at the time of pars plana vitrectomy (PPL group), and 25 patients who underwent phacoemulsification and IOL (MA60BM) implantation without vitrectomy (PE group). Intraocular lens tilt and decentration were evaluated quantitatively, using the anterior eye segment analysis system, approximately 12 months after surgery. RESULTS: There was no significant difference in IOL tilt (p = 0.47) or decentration (p = 0.26) between the PPL and PE groups. CONCLUSIONS: The present study suggests that tilt and decentration of the IOL are acceptable in combined vitrectomy and pars plana lensectomy. PMID- 16704705 TI - Ultraviolet radiation-B-induced cataract in albino rats: maximum tolerable dose and ascorbate consumption. AB - PURPOSE: To investigate the maximum tolerable dose (MTD) for cataract induced by ultraviolet radiation-B (UVB) in 7-week-old albino rats and to study the effect of UVB eye exposure on lens ascorbate content. METHODS: Fifty 7-week-old albino Sprague Dawley rats were unilaterally exposed in vivo to 300-nm UVB under anaesthesia, receiving 0, 0.25, 3.5, 4.3 and 4.9 kJ/m(2). The MTD was estimated based on lens forward light scattering measurements. Lens ascorbate content was determined in the processed lens using high performance liquid chromatography with UVR detection. RESULTS: Animals exposed to UVB doses >or=3.5 kJ/m(2) developed cortical cataracts. The MTD for avoidance of UVB-induced cataract was estimated to 3.01 kJ/m(2). UVB exposure decreased lens ascorbate concentration in the exposed lens in line with UVB dose, H(e), according to the models: C = C(NonCo) + C(Co)e(-kH(e) ) for exposed lenses; C = C(NonCo) + C(Co) for non exposed lenses, and C(d) = C(Co)(e(-kH(e) ) - 1). Parameters for consumable and non-consumable ascorbate were estimated to C(NonCo) = 0.04 and C(Co) = 0.11 micromol/g wet weight of lens. For lens ascorbate difference, tau = 1/k = 0.86 kJ/m(2). A total of 63% of UVB consumable ascorbate has been consumed after only tau = 0.86 kJ/m(2), while MTD(2.3 : 16) = 3.01 kJ/m(2), indicating that ascorbate decrease is in the order of 3.5 times more sensitive to detecting UVR damage in the lens than forward light scattering. CONCLUSIONS: The MTD for avoidance of UVB induced cataract in the 7-week-old albino Sprague Dawley rat was estimated to be 3.01 kJ/m(2). In vivo UVB exposure of the rat eye decreases lens ascorbate content following an exponential decline, and suprathreshold doses cause greater effect than subthreshold doses. PMID- 16704706 TI - Corneal transplantation in a developing country: problems associated with technology transfer from rich to poor societies. AB - PURPOSE: To evaluate the results of penetrating corneal transplantation at a university hospital in Brazil. METHODS: A prospective cohort study was conducted on 146 patients who underwent penetrating keratoplasty at the University Hospital, Faculty of Medicine of Ribeirao Preto, University of Sao Paulo, Brazil. The participants were followed postoperatively for 36 months or until graft failure. RESULTS: In this period, 49% of the grafts lost transparency. All the corneal diseases analysed were present in about the same proportions, except for keratoconus and bullous keratopathy, which showed 3% and 73% rates of graft oedema, respectively. Only 8% of the failures could be attributed to rejection. Other variables such as age, multiple surgeries and previous eye conditions could not explain the high rate of graft failure. Non-adherence to postoperative care increased with age, with a significant association between this behaviour and the rate of graft failure in the older age group. Keratoconus seems to be much less demanding and bullous keratopathy seems to be more exigent in terms of corneal transplantation than other corneal diseases. CONCLUSION: Although it involves an apparently simple surgical procedure, corneal transplantation is a demanding enterprise that can lead to very poor results under unfavourable conditions. PMID- 16704707 TI - Novel clinical application of sterilized, freeze-dried amniotic membrane to treat patients with pterygium. AB - PURPOSE: To evaluate the use of sterilized, freeze-dried amniotic membrane (FD AM) transplantation for pterygium surgery. METHODS: This study involved a prospective, non-comparative, interventional case series. Thirteen eyes of 13 patients with primary (eight eyes) or recurrent (five eyes) pterygium were studied. After excision of the pterygium fibrous tissues and application of intraoperative use of mitomycin-C, sterilized FD-AM was sutured over the bare scleral defect. The integrity of the FD-AM graft, epithelialization over the FD AM, pterygium recurrence and postoperative complications were evaluated. RESULTS: Postoperatively, the FD-AM was well retained in all patients, and complete epithelialization over the transplanted membrane was achieved within 1-2 weeks. All patients demonstrated early resolution of ocular inflammation and there was no recurrence of pterygium in any of the treated patients during the mean follow up of 13.9 +/- 6.0 months. No ocular complications were noted following transplantation. CONCLUSION: Sterilized FD-AM showed excellent biocompatibility on the human ocular surface. This novel and promising biomaterial may be a useful alternative to conjunctival grafting in the treatment of pterygium. PMID- 16704708 TI - Case series of 406 vernal keratoconjunctivitis patients: a demographic and epidemiological study. AB - PURPOSE: To evaluate the specific allergic sensitization and epidemiological characteristics of vernal keratoconjunctivitis (VKC). METHODS: This retrospective non-comparative case series included 406 VKC patients. Data included patient and family histories, and results of allergic tests. Annual incidence and prevalence rates were calculated for a cohort of 128 VKC patients from the greater Padua area. RESULTS: The great majority of VKC patients were male (76%), with a male : female ratio of 3.3 : 1. A skin prick test, specific serum IgE or conjunctival challenge was positive in 43%, 56% and 58% of patients, respectively. In the cohort of patients from the Padua area, the prevalence of the disease was 7.8/100,000, with a higher rate in young males (57/100,000) compared with young females (22/100,000), and lower rates in people over 16 years of age (3.8/100,000 in males, 1/100,000 in females). The incidence of VKC was 1/100,000, with a higher rate in males under 16 years of age (10/100,000) compared with females (4.2/100,000). In people over 16 years of age, the incidence of the disease was 0.06/100,000, with no difference between males and females. CONCLUSION: An IgE mediated sensitization was found in only half of the VKC patients. Vernal keratoconjunctivitis is not a rare event in the paediatric population but is an extremely rare new disease in adults. PMID- 16704709 TI - LASIK after retinal detachment surgery. AB - PURPOSE: To compare, in the same individuals, the safety and efficacy of laser in situ keratomileusis (LASIK) in eyes with and without previous retinal detachment surgery. METHODS: In a prospective clinical trial, seven myopic patients who had previously undergone scleral buckling surgery in one eye underwent conventional LASIK surgery in both eyes. Uncorrected visual acuity (UCVA), best corrected visual acuity (BCVA), refraction, Orbscan topography and pachymetry were recorded before and 1, 3, 6 and 12 months after surgery. The eyes were divided into two groups: group 1 consisted of eyes that had undergone previous surgery for retinal detachment, and group 2 consisted of the fellow eyes of the same patients, which had not undergone any previous ocular surgery. Student's t-test for match-paired data was used to evaluate the significance of differences. RESULTS: LASIK was performed successfully in all patients. The UCVA improved in all eyes in both groups. The mean change in the spherical equivalent between 1 and 12 months after LASIK surgery was 1.7 +/- 1.1 and 0.6 +/- 0.5 diopter in groups 1 and 2, respectively (p = 0.019). CONCLUSION: LASIK may be considered for treatment of myopia in eyes that have had previous surgery for retinal detachment. However, the risk of regression may be higher in such eyes than in eyes with no previous scleral buckling surgery. PMID- 16704710 TI - Isolated bilateral blindness as the sole manifestation of transient ischaemic attacks. AB - PURPOSE: To explain the rare phenomenon of acute transient bilateral blindness without additional ophthalmological or neurological symptoms and signs. METHODS: Six patients with isolated bilateral visual loss lasting 1-15 mins and occurring simultaneously in both eyes were evaluated. Clinical observation, neuroimaging (CT, MRI, MR-angiography), extra- and transcranial Doppler and vascular risk factors assessment were performed. RESULTS: Cortical blindness due to bilateral occipital lobe transient ischaemic attacks (TIAs) was established as the most likely cause of acute transient visual loss in all patients. CONCLUSION: We suggest that in cases of transient bilateral blindness with sudden onset, appearing simultaneously in both visual fields, a bilateral ischaemia of the visual cortex should be suspected even if other neurological symptoms are lacking. Patients should be managed in the same way as patients with vertebrobasilar TIAs. PMID- 16704711 TI - Magnetic resonance imaging determination of extraocular eye muscle volume in patients with thyroid-associated ophthalmopathy and proptosis. AB - PURPOSE: Recent studies concerning the association between extraocular muscle (EOM) enlargement in thyroid-associated ophthalmopathy (TAO) and immunological and clinical activity have not been conclusive, probably due to a lack of uniform imaging methods (ultrasonography, computer tomography [CT] or magnetic resonance imaging [MRI]) and difficulties in the determination of EOM volume. The aim of the present study was to examine the significance of EOM enlargement as established by MRI-based volume determination, with reference to proptosis and the presence of autoantibodies, clinical activity and the duration of active disease. METHODS: We determined EOM volume using MRI in 15 patients concomitantly with the determination of TSH, thyroid hormones, thyrotropin receptor antibodies (TRab) thyroid peroxidase antibodies (TPOab) and clinical activity score (CAS) at entry. We also established the duration until cessation of clinically active TAO. RESULTS: All 15 patients had bilateral EOM enlargement, but swelling of orbital fatty tissue was absent. Significant correlations between thickness of musculi rectales and proptosis, values of TRab, CAS, and duration of activity were observed. CONCLUSION: Our results support the hypothesis of a role of thyrotropin receptor antibodies in the pathogenesis of TAO and suggest that only EOM enlargement is responsible for proptosis in TAO. PMID- 16704712 TI - Post-streptococcal uveitis. AB - PURPOSE: To describe the clinical features of post-streptococcal uveitis (PSU) and examine management strategies in the treatment of this under-recognized condition. METHODS: Patients were identified from the world literature using the Pubmed search engine. We examined two new cases of post-streptococcal intermediate uveitis. The epidemiology, immune mechanisms, clinical features, investigations, treatments and visual outcomes were examined and recorded. RESULTS: We reviewed 11 patients including our own two cases. There was a statistically significant seasonal difference in antistreptolysin-O titres (ASOT), and age-related ASOT was identified. Of the 11 patients, eight (72.7%) had anterior uveitis, two (18.2%) had intermediate uveitis and one (9.1%) had panuveitis. Their ages ranged from 5 to 56 years (mean 17 years). The majority of cases had significantly elevated ASOT; most patients were treated with topical steroids and oral antibiotics and four cases underwent adenotonsillectomy. The visual prognosis was good in most cases. CONCLUSIONS: Uveitis may be the sole presenting clinical feature, or it may occur in combination with other features of post-streptococcal infection. Ophthalmologists should be aware of the clinical features of PSU and maintain a high level of suspicion, particularly in childhood uveitis. PMID- 16704713 TI - Exudation, response to photocoagulation and spontaneous remission in a case of bilateral racemose haemangioma. PMID- 16704714 TI - Inert intraocular eyelash foreign body following phacoemulsification cataract surgery. AB - INTRODUCTION: We describe the first reported case of tolerated eyelash inoculation following phacoemulsification surgery, with 4 years follow-up, that did not result in endophthalmitis or low grade chronic uveitis. CASE REPORT: A 79 year-old man with bilateral cataracts underwent phacoemulsification and posterior chamber intraocular lens implantation in the right eye. Three months later, he was found to have an eyelash within the right anterior chamber. The patient opted to leave his asymptomatic right eye alone. Four years later, he was referred to our hospital for review of his intraocular foreign body. He had remained asymptomatic with bilateral 6/6 vision, and without pain, photophobia or monocular diplopia. He declined surgery to remove the foreign body and opted for annual review. DISCUSSION: Intraocular implantation of cilia is a rare surgical complication following cataract surgery. The reaction of the eye to cilia has been reported to be variable and unpredictable, varying from absolute lack of reaction to endophthalmitis. Careful preoperative preparation can prevent similar incidents with potential complications. The position of the cilium is an important consideration. In contrast to other organic materials, a cilium is relatively inert and is rarely associated with infection. Acute inflammation most commonly occurs within days or after a few months, although the eye can tolerate the cilium for many years. Some ophthalmologists prefer observation, particularly when there is no inflammation present. We believe that each patient should be considered on an individual basis. The decision to remove an intraocular eyelash remains controversial and should be considered at the onset of inflammatory or infective clinical signs. PMID- 16704715 TI - Acute syphilitic posterior placoid chorioretinitis after an intravitreal triamcinolone acetonide injection. PMID- 16704717 TI - Psoriatic uveitis-associated cystoid macular oedema treated with intravitreal triamcinolone acetonide. PMID- 16704716 TI - Repeated intravitreal triamcinolone acetonide for chronic sympathetic ophthalmia. PMID- 16704718 TI - The internal limiting membrane can represent the roof of the foveal pseudocyst in a traumatized eye. PMID- 16704719 TI - Trans-zonular delivery of intravitreal triamcinolone acetonide in the management of pre-existing macular oedema during cataract surgery. PMID- 16704720 TI - The use of control charts in monitoring endophthalmitis. PMID- 16704721 TI - Does timolol LA enhance the disrupting effect of travoprost on the blood-aqueous barrier? PMID- 16704722 TI - Orbital pleomorphic lipoma in a 25-year-old male. PMID- 16704723 TI - Visual perceptions during cataract surgery. PMID- 16704724 TI - Efficacy of 2% lignocaine gel in pterygium surgery. PMID- 16704725 TI - Combined phacoemulsification and pars plana vitrectomy for macular hole treatment. PMID- 16704726 TI - Theory and practice--a case study of coordination and ownership in the Bangladesh health SWAp. AB - BACKGROUND: In the past decade the sector-wide approach (SWAp) model has been promoted by donors and adopted by governments in several countries. The purpose of this study is to look at how partners involved in the health SWAp in Bangladesh define ownership and coordination, in their daily work and to analyse the possible implications of these definitions. METHODOLOGY: The study object was a process of decision-making in the Government of Bangladesh in 2003. Information was collected through participant observations, interviews and document review. RESULTS: During the study period the Government of Bangladesh decided to reverse a decision to unify the two wings of the Ministry of Health and Family Welfare. The decision led to disagreements with development partners, which had serious implications for cooperation between key actors in the Bangladesh health sector leading to deteriorated relationships and suspension of donor funds. The donor community in itself was also in disagreement which led to inconsistencies in the dialogue between the development partners and the Government of Bangladesh. CONCLUSION: The case shows that main actors in the Bangladesh health SWAp interpret ownership and coordination, fundamental aspects of SWAp, differently. As long as work ran smoothly, the different definitions did not create any problems, but when disagreements arose they became an obstacle. It is concluded that partners in development should devote more effort to their working relationships and that responsibilities within a SWAp need to be more clearly delineated. PMID- 16704727 TI - Prediction of amphipathic in-plane membrane anchors in monotopic proteins using a SVM classifier. AB - BACKGROUND: Membrane proteins are estimated to represent about 25% of open reading frames in fully sequenced genomes. However, the experimental study of proteins remains difficult. Considerable efforts have thus been made to develop prediction methods. Most of these were conceived to detect transmembrane helices in polytopic proteins. Alternatively, a membrane protein can be monotopic and anchored via an amphipathic helix inserted in a parallel way to the membrane interface, so-called in-plane membrane (IPM) anchors. This type of membrane anchor is still poorly understood and no suitable prediction method is currently available. RESULTS: We report here the "AmphipaSeeK" method developed to predict IPM anchors. It uses a set of 21 reported examples of IPM anchored proteins. The method is based on a pattern recognition Support Vector Machine with a dedicated kernel. CONCLUSION: AmphipaSeeK was shown to be highly specific, in contrast with classically used methods (e.g. hydrophobic moment). Additionally, it has been able to retrieve IPM anchors in naively tested sets of transmembrane proteins (e.g. PagP). AmphipaSeek and the list of the 21 IPM anchored proteins is available on NPS@, our protein sequence analysis server. PMID- 16704728 TI - Explaining individual variation in patterns of mass loss in breeding birds. AB - BACKGROUND: Studies of birds have a disproportionate representation in the literature on life-history evolution, because of the (apparent) ease with which the costs and benefits can be quantified and manipulated. During reproduction, birds frequently show a highly conserved pattern of mass change and changes in mass loss during breeding have been widely considered to be a valid short-term measure of the costs of reproduction. Experimental manipulations of the breeding attempts of birds usually argue that the presence of a response shows that a cost of reproduction exists, but there is little consensus as to how the size of these costs can be measured. RESULTS: We model this mass loss by considering how a parent can maximise its lifetime reproductive success, using a theoretical framework that is particularly suited to modelling parental care in altricial birds. If lifetime reproductive success is taken to be the sum of a parent's current and future reproductive success, we show that the exact forms of these components will influence the optimal amount of mass a parent should lose. In particular, we demonstrate that the shape of the relationship between parental investment and chick survival will lead to differing degrees of investment between parents of different initial qualities: parents with initially high levels of energy reserves could conceivably invested a lesser, similar or greater amount of resources than parents with initially low reserves, and these initially 'heavy' parents could potentially end up being lighter than the initially 'lighter' individuals. CONCLUSION: We argue that it is difficult to make predictions about the dependence of a parent's final mass on its initial mass, and therefore mass loss should only be used as a short-term measure of the costs of reproduction with caution. The model demonstrates that we require a better understanding of the relationship between mass loss and both current and future reproductive success of the parent, before predictions about mass loss can be made and tested. We discuss steps that could be taken to increase the accuracy of our predictions. PMID- 16704729 TI - Characterization of the family of Mistic homologues. AB - BACKGROUND: Mistic is a unique Bacillus subtilis protein with virtually no detectable homologues in GenBank, which appears to integrate into the bacterial membrane despite an overall hydrophilic composition. These unusual properties have been shown to be useful for high-yield recombinant expression of other membrane proteins through fusion to the C-terminus of Mistic. To better understand the structure and function of Mistic, we systematically searched for and characterized homologous proteins among closely related bacteria. RESULTS: Three homologues of Mistic were found with 62% to 93% residue identity, all only 84 residues in length, corresponding to the C-terminal residues of B. subtilis Mistic. In every case, the Mistic gene was found partially overlapping a downstream gene for a K+ channel protein. Residue variation amongst these sequences is restricted to loop regions of the protein's structure, suggesting that secondary structure elements and overall fold have been conserved. Additionally, all three homologues retain the functional ability to chaperone fusion partners to the membrane. CONCLUSION: The functional core of Mistic consists of 84 moderately conserved residues that are sufficient for membrane targeting and integration. Understanding the minimal structural and chemical complexity of Mistic will lead to insights into the mechanistic underpinnings of Mistic-chaperoned membrane integration, as well as how to optimize its use for the recombinant heterologous expression of other integral membrane proteins of interest. PMID- 16704731 TI - Melatonin effects on luteinizing hormone in postmenopausal women: a pilot clinical trial NCT00288262. AB - BACKGROUND: In many mammals, the duration of the nocturnal melatonin elevation regulates seasonal changes in reproductive hormones such as luteinizing hormone (LH). Melatonin's effects on human reproductive endocrinology are uncertain. It is thought that the same hypothalamic pulse generator may both trigger the pulsatile release of GnRH and LH and also cause hot flashes. Thus, if melatonin suppressed this pulse generator in postmenopausal women, it might moderate hot flashes. This clinical trial tested the hypothesis that melatonin could suppress LH and relieve hot flashes. METHODS: Twenty postmenopausal women troubled by hot flashes underwent one week of baseline observation followed by 4 weeks of a randomized controlled trial of melatonin or matched placebo. The three randomized treatments were melatonin 0.5 mg 2.5-3 hours before bedtime, melatonin 0.5 mg upon morning awakening, or placebo capsules. Twelve of the women were admitted to the GCRC at baseline and at the end of randomized treatment for 24-hour sampling of blood for LH. Morning urine samples were collected twice weekly to measure LH excretion. Subjective responses measured throughout baseline and treatment included sleep and hot flash logs, the CESD and QIDS depression self-ratings, and the SAFTEE physical symptom inventory. RESULTS: Urinary LH tended to increase from baseline to the end of treatment. Contrasts among the 3 randomized groups were statistically marginal, but there was relative suppression combining the groups given melatonin as contrasted to the placebo group (p < 0.01 one-tailed, Mann-Whitney U = 14). Similar but not significant results were seen in blood LH. There were no significant contrasts among groups in hot flashes, sleep, depression, or side-effect measures and no significant adverse effects of any sort. CONCLUSION: The data are consistent with the hypothesis that melatonin suppresses LH in postmenopausal women. An effect related to the duration of nocturnal melatonin elevation is suggested. Effects of melatonin on reproductive endocrinology should be studied further in younger women and in men. Larger studies of melatonin effects on postmenopausal symptoms would be worthwhile. PMID- 16704730 TI - Functional studies of BCL11A: characterization of the conserved BCL11A-XL splice variant and its interaction with BCL6 in nuclear paraspeckles of germinal center B cells. AB - BACKGROUND: Chromosomal aberrations of BCL11A at 2p16.1 have been reported in a variety of B-cell malignancies and its deficiency in mice leads to a profound block in B-cell development. RESULTS: Alternative pre-mRNA splicing of BCL11A produces multiple isoforms sharing a common N-terminus. The most abundant isoform we have identified in human lymphoid samples is BCL11A-XL, the longest transcript produced at this locus, and here we report the conservation of this major isoform and its functional characterization. We show that BCL11A-XL is a DNA-sequence specific transcriptional repressor that associates with itself and with other BCL11A isoforms, as well as with the BCL6 proto-oncogene. Western blot data for BCL11A-XL expression coupled with data previously published for BCL6 indicates that these genes are expressed abundantly in germinal-center-derived B cells but that expression is extinguished upon terminal differentiation to the plasma cell stage. Although BCL11A-XL/BCL6 interaction can modulate BCL6 DNA binding in vitro, their heteromeric association does not alter the homomeric transcriptional properties of either on model reporter activity. BCL11A-XL partitions into the nuclear matrix and colocalizes with BCL6 in nuclear paraspeckles. CONCLUSION: We propose that the conserved N-terminus of BCL11A defines a superfamily of C2HC zinc-finger transcription factors involved in hematopoietic malignancies. PMID- 16704733 TI - Whose data set is it anyway? Sharing raw data from randomized trials. AB - BACKGROUND: Sharing of raw research data is common in many areas of medical research, genomics being perhaps the most well-known example. In the clinical trial community investigators routinely refuse to share raw data from a randomized trial without giving a reason. DISCUSSION: Data sharing benefits numerous research-related activities: reproducing analyses; testing secondary hypotheses; developing and evaluating novel statistical methods; teaching; aiding design of future trials; meta-analysis; and, possibly, preventing error, fraud and selective reporting. Clinical trialists, however, sometimes appear overly concerned with being scooped and with misrepresentation of their work. Both possibilities can be avoided with simple measures such as inclusion of the original trialists as co-authors on any publication resulting from data sharing. Moreover, if we treat any data set as belonging to the patients who comprise it, rather than the investigators, such concerns fall away. CONCLUSION: Technological developments, particularly the Internet, have made data sharing generally a trivial logistical problem. Data sharing should come to be seen as an inherent part of conducting a randomized trial, similar to the way in which we consider ethical review and publication of study results. Journals and funding bodies should insist that trialists make raw data available, for example, by publishing data on the Web. If the clinical trial community continues to fail with respect to data sharing, we will only strengthen the public perception that we do clinical trials to benefit ourselves, not our patients. PMID- 16704732 TI - Cell-type specific gene expression profiles of leukocytes in human peripheral blood. AB - BACKGROUND: Blood is a complex tissue comprising numerous cell types with distinct functions and corresponding gene expression profiles. We attempted to define the cell type specific gene expression patterns for the major constituent cells of blood, including B-cells, CD4+ T-cells, CD8+ T-cells, lymphocytes and granulocytes. We did this by comparing the global gene expression profiles of purified B-cells, CD4+ T-cells, CD8+ T-cells, granulocytes, and lymphocytes using cDNA microarrays. RESULTS: Unsupervised clustering analysis showed that similar cell populations from different donors share common gene expression profiles. Supervised analyses identified gene expression signatures for B-cells (427 genes), T-cells (222 genes), CD8+ T-cells (23 genes), granulocytes (411 genes), and lymphocytes (67 genes). No statistically significant gene expression signature was identified for CD4+ cells. Genes encoding cell surface proteins were disproportionately represented among the genes that distinguished among the lymphocyte subpopulations. Lymphocytes were distinguishable from granulocytes based on their higher levels of expression of genes encoding ribosomal proteins, while granulocytes exhibited characteristic expression of various cell surface and inflammatory proteins. CONCLUSION: The genes comprising the cell-type specific signatures encompassed many of the genes already known to be involved in cell-type specific processes, and provided clues that may prove useful in discovering the functions of many still unannotated genes. The most prominent feature of the cell type signature genes was the enrichment of genes encoding cell surface proteins, perhaps reflecting the importance of specialized systems for sensing the environment to the physiology of resting leukocytes. PMID- 16704734 TI - Analysis of right anterolateral impacts: the effect of trunk flexion on the cervical muscle whiplash response. AB - BACKGROUND: The cervical muscles are considered a potential site of whiplash injury, and there is a need to understand the cervical muscle response under non conventional whiplash impact scenarios, including variable body position and impact direction. There is no data, however, on the effect of occupant position on the muscle response to frontal impacts. Therefore, the objective of the study was to measure cervical muscle response to graded right anterolateral impacts. METHODS: Twenty volunteers were subjected to right anterolateral impacts of 4.3, 7.8, 10.6, and 12.8 m/s(2) acceleration with their trunk flexed forward 45 degrees and laterally flexed right or left by 45 degrees. Bilateral EMG of the sternocleidomastoids, trapezii, and splenii capitis and acceleration of the sled, torso, and head were measured. RESULTS AND DISCUSSION: With either direction of trunk flexion at impact, the trapezius EMGs increased with increasing acceleration (p < 0.05). Time to onset of the electromyogram and time to peak electromyogram for most muscles showed a trend towards decreasing with increasing acceleration. With trunk flexion to the left, the left trapezius generated 38% of its maximal voluntary contraction (MVC) EMG, while the right trapezius generated 28% of its MVC EMG. All other muscles generated 25% or less of this measure (25% for the left splenius capitis, 8% for the right splenius capitis, 6% for the left sternocleidomastoid, and 2% for the left sterncleidomastoid). Conversely, with the trunk flexed to the right, the right trapezius generated 44% of its MVC EMG, while the left trapezius generated 31% of this value, and all other muscles generated 20% or less of their MVC EMG (20% for the left splenius capitis, 14% for the right splenius capitis, 4% for both the left and right sternocleidomastoids). CONCLUSION: When the subject sits with trunk flexed out of neutral posture at the time of anterolateral impact, the cervical muscle response is dramatically reduced compared to frontal impacts with the trunk in neutral posture. In the absence of bodily impact, the flexed trunk posture appears to produce a biomechanical response that would decrease the likelihood of cervical muscle injury in low velocity impacts. PMID- 16704735 TI - Artemether-lumefantrine versus artesunate plus amodiaquine for treating uncomplicated childhood malaria in Nigeria: randomized controlled trial. AB - BACKGROUND: The therapeutic efficacy of artesunate plus amodiaquine and artemether/lumefantrine were assessed in an area of Nigeria with high levels of Plasmodium falciparum resistance to chloroquine and sulphadoxine-pyrimethamine. PARTICIPANTS: Children aged 6 to 59 months with uncomplicated P. falciparum infection and parasite density 1,000 to 200,000 parasites/microL enrolled following informed consent by parents. METHODS: Eligible children were randomly assigned to receive either a 3-day course of artesunate (4 mg/kg) plus amodiaquine (10 mg/kg) or 6-dose course of artemether/lumefantrine (20/120 mg tablets) over three days. Patients were followed up with clinical and laboratory assessments until day 14 using standard WHO in-vivo antimalarial drug test protocol. RESULTS: A total 119 eligible children were enrolled but 111 completed the study. Adequate clinical and parasitological response (ACPR) was 47 (87.0%) and 47 (82.5%) for artemether-lumefantrine (AL) and artesunate+amodiaquine (AAMQ) respectively (OR 0.7, 95% confidence interval 0.22 to 2.22). Early treatment failure (ETF) occurred in one participant (1.8%) treated with AAQ but in none of those with AL. Two (3.7%) patients in the AL group and none in the AAQ group had late clinical failure. Late parasitological failure was observed in 9 (15.8) and 5 (9.3%) of patients treated with AAQ and AL respectively. None of participants had a serious adverse event. CONCLUSION: Artemether-lumenfantrine and artesunate plus amodiaquine have high and comparable cure rates and tolerability among under five children in Calabar, Nigeria. PMID- 16704736 TI - A novel method to measure regional muscle blood flow continuously using NIRS kinetics information. AB - BACKGROUND: This article introduces a novel method to continuously monitor regional muscle blood flow by using Near Infrared Spectroscopy (NIRS). We demonstrate the feasibility of the new method in two ways: (1) by applying this new method of determining blood flow to experimental NIRS data during exercise and ischemia; and, (2) by simulating muscle oxygenation and blood flow values using these newly developed equations during recovery from exercise and ischemia. METHODS: Deoxy (Hb) and oxyhemoglobin (HbO2), located in the blood of the skeletal muscle, carry two internal relationships between blood flow and oxygen consumption. One is a mass transfer principle and the other describes a relationship between oxygen consumption and Hb kinetics in a two-compartment model. To monitor blood flow continuously, we transfer these two relationships into two equations and calculate the blood flow with the differential information of HbO2 and Hb. In addition, these equations are used to simulate the relationship between blood flow and reoxygenation kinetics after cuff ischemia and a light exercise. Nine healthy subjects volunteered for the cuff ischemia, light arm exercise and arm exercise with cuff ischemia for the experimental study. RESULTS: Analysis of experimental data of both cuff ischemia and light exercise using the new equations show greater blood flow (four to six times more than resting values) during recovery, agreeing with previous findings. Further, the simulation and experimental studies of cuff ischemia and light exercise agree with each other. CONCLUSION: We demonstrate the accuracy of this new method by showing that the blood flow obtained from the method agrees with previous data as well as with simulated data. We conclude that this novel continuous blood flow monitoring method can provide blood flow information non-invasively with NIRS. PMID- 16704737 TI - Predicting outbreaks: a spatial risk assessment of West Nile virus in British Columbia. AB - BACKGROUND: West Nile virus (WNv) has recently emerged as a health threat to the North American population. After the initial disease outbreak in New York City in 1999, WNv has spread widely and quickly across North America to every contiguous American state and Canadian province, with the exceptions of British Columbia (BC), Prince Edward Island and Newfoundland. In this study we develop models of mosquito population dynamics for Culex tarsalis and C. pipiens, and create a spatial risk assessment of WNv prior to its arrival in BC by creating a raster based mosquito abundance model using basic geographic and temperature data. Among the parameters included in the model are spatial factors determined from the locations of BC Centre for Disease Control mosquito traps (e.g., distance of the trap from the closest wetland or lake), while other parameters were obtained from the literature. Factors not considered in the current assessment but which could influence the results are also discussed. RESULTS: Since the model performs much better for C. tarsalis than for C. pipiens, the risk assessment is carried out using the output of C. tarsalis model. The result of the spatially-explicit mosquito abundance model indicates that the Okanagan Valley, the Thompson Region, Greater Vancouver, the Fraser Valley and southeastern Vancouver Island have the highest potential abundance of the mosquitoes. After including human population data, Greater Vancouver, due to its high population density, increases in significance relative to the other areas. CONCLUSION: Creating a raster-based mosquito abundance map enabled us to quantitatively evaluate WNv risk throughout BC and to identify the areas of greatest potential risk, prior to WNv introduction. In producing the map important gaps in our knowledge related to mosquito ecology in BC were identified, as well, it became evident that increased efforts in bird and mosquito surveillance are required if more accurate models and maps are to be produced. Access to real time climatic data is the key for developing a real time early warning system for forecasting vector borne disease outbreaks, while including social factors is important when producing a detailed assessment in urban areas. PMID- 16704738 TI - Identification of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-inducible genes in human amniotic epithelial cells. AB - BACKGROUND: Exposure to dioxins results in a broad range of pathophysiological disorders in human fetuses. In order to evaluate the effects of dioxins on the feto-placental tissues, we analyzed the gene expression in 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) treated primary cultures of human amniotic epithelial cells. METHODS: Human amniotic epithelial cells were dispersed by trypsin from amniotic membranes and cultured in DME/Ham's F12 medium supplemented with 10% FBS. Two weeks after plating, cells were treated with 50 nM TCDD or DMSO (control), further incubated for 48 hrs, and the gene expression was analyzed by DNA microarray technology and quantitative real-time PCR. RESULTS: Thirty eight TCDD-inducible genes, including cytochromeP4501A1 and cytochromeP4501B1, were identified. One of the remarkable profiles of the gene expression was the prominent up-regulation of interferon-inducible genes. The genes involved in the interferon gene expression and interferon signaling pathways were also up regulated. Furthermore, the expression of genes related to collagen synthesis or degradation was enhanced by TCDD. CONCLUSION: Using DNA microarray and quantitative real-time PCR analyses, we identified TCDD-inducible genes, including interferon-inducible genes and genes related to collagen synthesis or degradation, in human amniotic epithelial cells. PMID- 16704739 TI - Splenic and portal vein thrombosis in pancreatic metastasis from renal cell carcinoma. AB - BACKGROUND: Pancreatic metastases from previously treated renal cell carcinoma are uncommon. Surgical resection of pancreatic metastasis remains the only worthwhile modality of treatment. CASE PRESENTATION: A case where pancreatic metastasis from previously resected right sided renal cell carcinoma was resected with a subtotal left pancreatectomy is described. An unusual feature was the presence of a large splenic vein tumor thrombus extending into the portal vein with associated portal hypertension. The patient underwent an uneventful portal vein resection with primary anastomosis. CONCLUSION: This is possibly the first documented case of portal vein renal tumor thrombosis in a case of isolated pancreatic metastasis from previously operated renal cell carcinoma in published world surgical literature. PMID- 16704740 TI - Characterization of human malignant mesothelioma cell lines orthotopically implanted in the pleural cavity of immunodeficient mice for their ability to grow and form metastasis. AB - BACKGROUND: Malignant pleural mesothelioma (MPM) is a tumor known to be resistant to conventional therapies. Thus, an in vivo model can represent an important tool for assessing the efficacy of novel approaches in the treatment of MPM.Presently, human MPM cells have been grown orthotopically in mice upon transplantation of tumor masses or tumor cell suspensions following surgery. In these models however, surgery can interfere with the tumor growth and the early stages of tumor development cannot be easily explored. Finally, results may not be so accurate due to implantation of potentially different tumor samples in different experimental groups.Our work aimed at establishing a nude mouse model xenotransplanted with human MPM cell lines in which tumor progression exhibits some features of the human disease. METHODS: Three distinct human MPM cell lines previously established from MPM patients displaying two different phenotypes, biphasic (MM-B1 and IST-Mes3) and epithelioid (IST-Mes2), were directly injected into the pleural cavity of nude mice. At different times, mice were sacrificed for autopsy, tumor nodules were counted and then removed for histology. Presence of metastases in visceral organs was also monitored. RESULTS: IST-Mes2 cells were unable to grow in nude mice. MM-B1 and IST-Mes3 cells were capable of growing in nude mice and formed tumor nodules in the pleura. Post-mortem examination showed that MPM cells progressively colonized the parietal and visceral pleura, the diaphragm, the mediastinum and, lastly the lung parenchyma. No pneumo-thorax was evidenced in the mice. Pleural effusions as well as lymph node metastases were observed only at later times. CONCLUSION: This model mimics the progression of human malignant mesothelioma and it is easy to perform and reproducible; therefore it can be useful to study human MPM biology and evaluate the efficacy of novel therapies. PMID- 16704741 TI - Helicobacter pylori infection is not associated with an increased hemorrhagic risk in patients in the intensive care unit. AB - INTRODUCTION: The potential role of Helicobacter pylori in acute stress ulcer in patients in an intensive care unit (ICU) is controversial. The aim of this study was to determine the frequency of H. pylori infection in ICU patients by antigen detection on rectal swabs, and to analyze the potential relationship between the presence of H. pylori and the risk of digestive gastrointestinal bleeding. METHODS: In this prospective, multicenter, epidemiological study, the inclusion criteria were as follows: patients admitted to the 12 participating ICU for at least two days, who were free of hemorrhagic shock and did not receive more than four units of red blood cells during the day before or the first 48 hours after admission to the ICU. Rectal swabs were obtained within the first 24 hours of admission to the ICU and were tested for H. pylori antigens with the ImmunoCard STAT! HpSA kit. The following events were analyzed according to H. pylori status: gastrointestinal bleeding, unexplained decline in hematocrit, and the number of red cell transfusions. RESULTS: The study involved 1,776 patients. Forty-nine patients (2.8%) had clinical evidence of upper digestive bleeding. Esophagogastroduodenoscopy was performed in 7.6% of patients. Five hundred patients (28.2%) required blood transfusion. H. pylori antigen was detected in 6.3% of patients (95% confidence interval 5.2 to 7.5). H. pylori antigen positivity was associated with female sex (p < 0.05) and with a higher Simplified Acute Physiology Score II (SAPS II; p < 0.05). H. pylori antigen status was not associated with the use of fiber-optic gastroscopy, the need for red cell transfusions, or the number of red cell units infused. CONCLUSION: This large study reported a small percentage of H. pylori infection detected with rectal swab sampling in ICU patients and showed that the patients infected with H. pylori had no additional risk of gastrointestinal bleeding. Thus H. pylori does not seem to have a major role in the pathogenesis of acute stress ulcer in ICU patients. PMID- 16704743 TI - The incidence and outcome of septic shock patients in the absence of early-goal directed therapy. AB - INTRODUCTION: The purpose of the present study was to measure the incidence and outcome of septic patients presenting at the emergency department (ED) with criteria for early goal-directed therapy (EGDT). METHOD: This hospital-based, retrospective, observational study using prospectively collected electronic databases was based in a teaching hospital in Melbourne, Australia. We conducted outcome-blinded electronic screening of patients with infection admitted via the ED from 1 January 2000 to 30 June 2003. We obtained data on demographics, laboratory and clinical features on admission. We used paper records to confirm electronic identification of candidates for EGDT and to study their treatment. We followed up all patients until hospital discharge or death. RESULTS: Of 4,784 ED patients with an infectious disease diagnosis, only 50 fulfilled published clinical inclusion criteria for EGDT (EGDT candidates). Of these patients, 37 (74%) survived their hospital admission, two (4%) died in the ED, eight (16%) died in the intensive care unit and three (6%) died in the ward. After review of all ward cardiac arrests and non-NFR ('not for resuscitation') ward deaths, we identified a further two potential candidates for EGDT for an overall mortality of 28.8% (15 out of 52 patients). Analysis of treatment showed that twice as many (70%) of the EGDT candidates received vasopressor therapy in the ED, and their initial mean central venous pressure (10.8 mmHg) was almost twice that in patients from the EGDT study conducted by Rivers and coworkers. CONCLUSION: In an Australian teaching hospital candidates for EGDT were uncommon and, in the absence of an EGDT protocol, their mortality was lower than that reported with EGDT. PMID- 16704742 TI - Empiric broad-spectrum antibiotic therapy of nosocomial pneumonia in the intensive care unit: a prospective observational study. AB - INTRODUCTION: Antibiotic de-escalation, which consists of the initial institution of empiric broad-spectrum antibiotics followed by antibiotic streamlining driven by microbiological documentation, is thought to provide maximum benefit for the individual patient, while reducing the selection pressure for resistance. METHODS: To assess a carbapenem-based de-escalating strategy in nosocomial pneumonia (NP), a prospective observational study was conducted in critically ill patients with NP treated empirically with imipenem +/- aminoglycoside/glycopeptide in 24 intensive care units of Spanish general hospitals. Overall, 244 patients were assessable (91% with late-onset NP). The primary outcome was therapeutic success 7-9 days post therapy. RESULTS: Microbial identification--based on cultures of tracheal aspirates in 82% of patients, cultures of protected specimen brush in 33%, and cultures of bronchoalveolar lavage in 4%--was only available for 131 (54%) patients. Initial antibiotics were inadequate for 23 (9%) patients. Of the remaining patients, antibiotics were streamlined in 56 (23%) patients and remained unchanged in 14 (6%) patients based on microbiology data, in 38 (16%) patients despite microbiology data favouring de escalation, and in 113 (46%) patients due to unknown aetiology. Overall, de escalation was implemented in only 23% of patients with potentially multiresistant pathogens, compared with 68% of patients with the remaining pathogens (P < 0.001). Response rates were 53% for patients continuously treated with imipenem-based regimens and 50% for the de-escalated patients. Higher Acute Physiology, Age, and Chronic Health Evaluation II scores were associated with greater mortality, whereas adequate empiric antibiotic therapy protected against fatal outcomes. No increase of superinfection rates caused by emerging pathogens was observed. The costs associated with de-escalation were mainly dependent on the duration of hospitalization. CONCLUSION: This study mainly highlights the current practice of a specific algorithm of de-escalation solely based on the available microbiological data, and highlights the barriers to using it more widely. In this setting, de-escalation was less likely to occur in the presence of potentially multiresistant pathogens. Prior antibiotic administration and the low use of bronchoscopic techniques may have influenced negatively the implementation of de-escalation. Optimization of de-escalation strategies for NP should rely on a correct choice of empiric antibiotics, on appropriate microbiological investigations, and on a balanced interpretation of microbiological and clinical data. PMID- 16704745 TI - Human, viral or mutant human IL-10 expressed after local adenovirus-mediated gene transfer are equally effective in ameliorating disease pathology in a rabbit knee model of antigen-induced arthritis. AB - IL-10 is a Th2 cytokine important for inhibiting cell-mediated immunity while promoting humoral responses. Human IL-10 (hIL-10) has anti-inflammatory, immunosuppressive as well as immunostimulatory characteristics, whereas viral IL 10 (vIL-10), a homologue of hIL-10 encoded by Epstein Barr virus (EBV), lacks several immunostimulatory functions. The immunostimulatory characteristic of hIL 10 has been attributed to a single amino acid, isoleucine at position 87, which in vIL-10 is alanine. A mutant hIL-10 in which isoleucine has been substituted (mut.hIL-10) is biologically active with only immunosuppressive, but not immunostimulatory, functions, making it a potentially superior therapeutic for inflammatory diseases. To compare the efficacy of mut.hIL-10 with hIL-10 and vIL 10 in blocking the progression of rheumatoid arthritis, we used replication defective adenoviral vectors to deliver intra-articularly the gene encoding hIL 10, vIL-10 or mut.hIL-10 to antigen-induced arthritic (AIA) knee joints in rabbits. Intra-articular expression of hIL-10, vIL-10, and mut.hIL-10 resulted in significant improvement of the pathology in the treated joints to similar levels. These observed changes included a significant reduction in intra-articular leukocytosis and the degree of synovitis, as well as normalization of cartilage matrix metabolism. Our results suggest that hIL-10, vIL-10, and mut.hIL-10 are all equally therapeutic in the rabbit AIA model for treating disease pathology. PMID- 16704744 TI - Inhibition of macropinocytosis blocks antigen presentation of type II collagen in vitro and in vivo in HLA-DR1 transgenic mice. AB - Professional antigen-presenting cells, such as dendritic cells, macrophages and B cells have been implicated in the pathogenesis of rheumatoid arthritis, constituting a possible target for antigen-specific immunotherapy. We addressed the possibility of blocking antigen presentation of the type II collagen (CII) derived immunodominant arthritogenic epitope CII259-273 to specific CD4 T cells by inhibition of antigen uptake in HLA-DR1-transgenic mice in vitro and in vivo. Electron microscopy, confocal microscopy, subcellular fractionation and antigen presentation assays were used to establish the mechanisms of uptake, intracellular localization and antigen presentation of CII by dendritic cells and macrophages. We show that CII accumulated in membrane fractions of intermediate density corresponding to late endosomes. Treatment of dendritic cells and macrophages with cytochalasin D or amiloride prevented the intracellular appearance of CII and blocked antigen presentation of CII259-273 to HLA-DR1 restricted T cell hybridomas. The data suggest that CII was taken up by dendritic cells and macrophages predominantly via macropinocytosis. Administration of amiloride in vivo prevented activation of CII-specific polyclonal T cells in the draining popliteal lymph nodes. This study suggests that selective targeting of CII internalization in professional antigen-presenting cells prevents activation of autoimmune T cells, constituting a novel therapeutic strategy for the immunotherapy of rheumatoid arthritis. PMID- 16704748 TI - Suicide in Hong Kong: a case-control psychological autopsy study. AB - BACKGROUND: The relative contribution of psychosocial and clinical risk factors to suicide among Chinese populations is an important issue. In Hong Kong, this issue requires vigorous examination in light of a 50% increase in suicide rate between 1997 and 2003. METHOD: Using a case-control psychological autopsy method, 150 suicide deceased were compared with 150 living controls matched by age and gender. Semi-structured interviews were conducted with the next-of-kin of the subjects. Data were collected on a wide range of potential risk and protective factors, including demographic, life event, clinical and psychological variables. The relative contribution of these factors towards suicide was examined in a multiple logistic regression model. RESULTS: Six factors were found to significantly and independently contribute to suicide: unemployment, indebtedness, being single, social support, psychiatric illness, and history of past attempts. CONCLUSIONS: Both psychosocial and clinical factors are important in suicides in Hong Kong. They seem to have mediated suicide risk independently. In addition, socio-economic adversities seem to have played a relatively important role in the increasing suicide rate in Hong Kong. PMID- 16704746 TI - Knee cartilage loss in symptomatic knee osteoarthritis over 4.5 years. AB - The objective of this study was to describe the rate of change in knee cartilage volume over 4.5 years in subjects with symptomatic knee osteoarthritis (OA) and to determine factors associated with cartilage loss. One hundred and five subjects were eligible for this longitudinal study. Subjects' tibial cartilage volume was assessed by magnetic resonance imaging (MRI) at baseline, at 2 years and at 4.5 years. Of 105 subjects, 78 (74%) completed the study. The annual percentage losses of medial and lateral tibial cartilage over 4.5 years were 3.7 +/- 4.7% (mean +/- SD; 95% confidence interval 2.7 to 4.8%) and 4.4 +/- 4.7% (mean +/- SD; 95% confidence interval 3.4 to 5.5%), respectively. Cartilage volume in each individual seemed to track over the study period, relative to other study participants. After multivariate adjustment, annual medial tibial cartilage loss was predicted by lesser severity of baseline knee pain but was independent of age, body mass index and structural factors. No factors specified a priori were associated with lateral cartilage volume rates of change. Tibial cartilage declines at an average rate of 4% per year in subjects with symptomatic knee OA. There was evidence to support the concept that tracking occurs in OA. This may enable the prediction of cartilage change in an individual. The only significant factor affecting the loss of medial tibial cartilage was baseline knee pain, possibly through altered joint loading. PMID- 16704749 TI - Functional MRI correlates of the recall of unresolved life events in borderline personality disorder. AB - BACKGROUND: Patients with borderline personality disorder (BPD) frequently report unresolved life events but it is still poorly understood, how these experiences are represented in the brain. Using functional magnetic resonance imaging (fMRI), the present study aimed at investigating the neural correlates of the recall of unresolved life events in patients with BPD and healthy controls. METHOD: Twenty female BPD patients and 21 healthy control subjects underwent fMRI. During measurement subjects recalled unresolved and resolved negative life events. Individual cue words were used to stimulate autobiographical memory. After scanning, subjects rated their emotional states during the recall of both types of memories. RESULTS: When contrasting unresolved and resolved life events, patients showed significant bilateral activation of frontotemporal areas including the insula, amygdala, and the anterior cingulate cortex, the left posterior cingulate cortex, right occipital cortex, the bilateral cerebellum and the midbrain. In healthy subjects, no differential brain activation was related to these conditions. The 2 x 2 factorial analysis (DeltaBPD - Deltacontrols) revealed similar results with bilateral activation of the frontal cortex including parts of the insula and of the orbitofrontal cortex, temporal activation including the amygdala, activation of the right occipital cortex, and parts of the cerebellum. Patients but not controls reported higher levels of anxiety and helplessness during the unresolved versus resolved memory condition. CONCLUSIONS: The activation of both, the amygdala and prefrontal areas, might reflect an increased effortful but insufficient attempt to control intensive emotions during the recall of unresolved life events in patients with BPD. PMID- 16704750 TI - Whither clinical validation? PMID- 16704751 TI - Clinical and laboratory aspects of thyroid autoantibodies. AB - This review describes the aetiology of the major thyroid antigens. Iodination of thyroglobulin produces multiple antigen configurations which are functionally active but immunologically distinct. The thyroid stimulating hormone (TSH) receptor is a two-subunit glycoprotein; the extracellular A subunit is recognized by thyroid stimulating antibodies, while those antibodies recognizing the B subunit, located much nearer the cell surface, appear to function as blocking antibodies. Thyroid peroxidase (TPO), originally described as thyroid microsomal antigen, is present on the apical surface of thyroid follicular cells and is the antigen involved in cell-mediated cytotoxicity. Multiple B-cell-reactive epitopes exist, each giving rise to different antibodies. The aetiology and mechanics of the autoimmune cellular and antibody responses involves a combination of human leucocyte antigen (HLA) linkage, genetics and environmental factors to determine the initial and subsequent stages of the development of autoimmune thyroid disease. Depending on the antibody, a combination of enzyme-linked immunosorbent assay for TPO and thyroglobulin and bioassays and/or radioimmunoassay for TSH receptor antibodies are used to estimate their concentrations. The other conditions with which autoimmune thyroid diseases are associated include, for example, pernicious anaemia, connective tissue disorders, diabetes, coeliac disease, mood disorders like depression and fertility-related problems such as miscarriage, infertility, in vitro fertilization failure, pre-term delivery and postpartum thyroiditis. Often, there is no cause-and-effect relationship between them and it is debatable in some cases whether it is worthwhile monitoring patients with autoimmune thyroid disease for other conditions or vice versa. The review also itemizes the circumstances in which it might be useful to measure each antibody (i.e. the use of TPO antibodies in investigation of goitre, diagnosis of Graves' and Hashimoto's disease and the prediction of risk of developing hypothyroidism during subclinical thyroid disease; TSH receptor antibodies in maternal and neonatal hyperthyroidism and thyroglobulin antibodies in the monitoring and treatment of thyroid carcinoma). Finally, taking the current literature into account, an algorithm is recommended for the most effective use of these antibodies in the investigation of autoimmune thyroid disease. PMID- 16704752 TI - The effect of thyroid dysfunction on N-terminal pro-B-type natriuretic peptide concentrations. AB - BACKGROUND: B-type natriuretic peptides and N-terminal pro-B-type natriuretic peptides (NT-proBNP) have been suggested as being useful for the diagnosis of congestive heart failure. We have shown previously that changes in thyroid function are associated with changes in concentrations of other low molecular weight molecules. Therefore, the aim of this study was to assess whether similar changes occurred with NT-proBNP concentrations following treatment of thyroid dysfunction. METHODS: Seventeen patients (12 female, 5 male, age range 24-77 years) with newly diagnosed hypothyroidism and 21 patients (16 female, 5 male, age range 21-66 years) with newly diagnosed hyperthyroidism had NT-proBNP measured at baseline and when they subsequently became euthyroid. RESULTS: NT proBNP levels were not significantly different in the hyperthyroid group from the hypothyroid, either before (P = 0.706) or after treatment to euthyroidism (P = 0.170). The hypothyroid group showed a rise in NT-proBNP after treatment (P < 0.001). There was a marginally significant fall in the NT-proBNP levels in the hyperthyroid group (P = 0.05). However, these changes were within the expected wide range of biological variability of NT-proBNP shown in earlier studies. CONCLUSIONS: Hypothyroidism alters NT-proBNP concentrations, but the magnitude of this effect may be lost in the wide biological variability of natriuretic peptides and this statistically significant finding is not likely to be of clinical relevance. PMID- 16704753 TI - Investigation into possible causes of interference in serum testosterone measurement in women. AB - BACKGROUND: Direct (non-extracted) testosterone immunoassays may give spuriously high results in women. The presumed interferents may be removed if testosterone is extracted into an organic phase before being measured. We aimed to clarify possible causes and effects of interference in testosterone measurement in women. METHODS: Women who had a blood sample referred to Hope Hospital Clinical Biochemistry laboratory for measurement of serum testosterone concentration over a six-month period were studied. Clinical and treatment data were recorded. A difference (direct-minus-extracted testosterone) of less than 1.0 nmol/L was used to define a group with low interference. A difference of 2.5 nmol/L or more was used to define a group with significant interference. RESULTS: The distribution of interference in 1271 serum samples from female patients was unimodal. There were no group differences in clinical presentation or treatment. The median degree of interference was 1.4 nmol/L. In 42 female patients with varying degrees of interference, identified on routine assay, regression analysis showed strong association between dehydroepiandrosterone sulphate (DHEA-S) and interference (direct-minus-extracted testosterone) (r = 0.77, P < 0.001). CONCLUSIONS: Given that DHEA-S is present in very variable amounts in blood, it is possible that even with a low cross reactivity, DHEA-S or one of its metabolites significantly interferes in the testosterone assay when at higher concentrations. PMID- 16704754 TI - Dehydroepiandrosterone sulphate interferes with the Abbott Architect direct immunoassay for testosterone. AB - BACKGROUND: There are known difficulties in measuring testosterone in women using 'direct' (non-extraction) immunoassays. The positive bias encountered in many of these assays has been attributed to an unidentified interfering substance. We have evaluated the potential for dehydroepiandrosterone sulphate (DHEA-S) to cross-react in the Abbott Architect testosterone assay. METHODS: Samples (n = 81) from female patients who had DHEA-S, direct and extracted testosterone measured were examined retrospectively. Possible cross-reactivity was evaluated by spiking both testosterone free assay diluent and serum from a woman with exogenous DHEA S. RESULTS: The retrospective patient analysis of the data demonstrated a relationship between interference and DHEA-S (r = 0.78, P < or = 0.0001). Spiking studies revealed a linear relationship between added DHEA-S and the measured testosterone concentration. Consistent with the retrospective study, for every 1 micromol/L increase in DHEA-S, 0.23 nmol/L of testosterone was measured in the diluent. Spiking DHEA-S into serum gave only a 0.12 nmol/L increase in measured testosterone for every 1 micromol/L rise in DHEA-S. CONCLUSION: DHEA-S cross reacts in the Abbott Architect direct immunoassay for testosterone to a clinically significant effect. This cross-reactivity is likely to be responsible for much of the interference observed when measuring testosterone in women by this method. PMID- 16704755 TI - Hyperhomocysteinaemia, methylenetetrahydrofolate reductase polymorphism and risk of coronary artery disease. AB - BACKGROUND: Hyperhomocysteinaemia is an independent, graded risk factor for coronary artery disease (CAD). The methylenetetrahydrofolate reductase (MTHFR) polymorphism is associated with hyperhomcysteinaemia and may therefore influence individual susceptibility to CAD. We have investigated this risk factor in a Tunisian Arab population. METHODS: Polymerase chain reaction-restriction fragment length polymorphism analysis was used to detect the C677T and A1298C variants of the MTHFR gene in 100 patients with CAD and 120 healthy controls. The severity of CAD was expressed as the number of affected vessels. Plasma total homocysteine (tHcy) concentration was determined using a direct chemiluminescence assay. RESULTS: MTHFR CC, CT and TT genotype frequencies in the CAD group were significantly different from those observed in the control group (49%, 35% and 16% versus 48.3%, 45.8% and 5.8%, respectively; P = 0.031). However, MTHFR AA, AC and CC genotypes frequencies in the CAD group were not significantly different from the control group ( P = 0.568). Patients with CAD showed higher plasma tHcy concentrations than patients without CAD (15.86 +/- 8.63 micromol/L versus 11.90 +/- 3.25 micromol/L, P < 0.001). There was no association between the MTHFR polymorphisms and the number of stenosed vessels. Patients with the MTHFR TT genotype had higher plasma tHcy, serum creatinine, cholesterol and triglyceride concentrations than patients with the MTHFR CC genotype. CONCLUSIONS: The C677T polymorphism of the MTHFR gene is associated with hyperhomocysteinaemia, lipid dysregulation and the presence of CAD in this Tunisian Arab population. PMID- 16704756 TI - The effect of alkaline phosphatase inhibitors on intracellular lipid accumulation in preadipocytes isolated from human mammary tissue. AB - BACKGROUND: A previous study has demonstrated that alkaline phosphatase (AP) may play a role in the control of intracellular lipid accumulation in the rodent preadipocyte cell line, 3T3-L1. The present study investigated whether AP may have a similar function in preadipocytes isolated from human mammary gland tissue. METHODS: Preadipocyte maturation was induced in the presence or absence of the tissue non-specific AP inhibitors levamisole and histidine, and the tissue specific AP inhibitor PheGlyGly. Cellular AP activity and adipogenesis were both assessed at 0 and 12 days post-induction of differentiation. RESULTS: After differentiation, AP activity increased 5.1 +/- 1.3-fold in the absence and 8.9 +/ 2.8-fold (P < 0.05) in the presence of levamisole. However, adipogenesis increased 1.95 +/- 0.11-fold in the absence but only 1.36 +/- 0.06-fold (P < 0.001) in the presence of levamisole. There was a 4.2 +/- 2.2-fold increase in AP activity in the absence and a 0.51 +/- 0.46-fold (P < 0.05) decrease in the presence of histidine. Adipogenesis increased 2.09 +/- 0.35-fold in the absence of histidine but only 1.22 +/- 0.30-fold (P < 0.05) in the presence of histidine. PheGlyGly had no effects. Fluorescent microscopy showed AP activity was localized to the triglyceride-containing droplets of the cell. CONCLUSION: This is the first study to show that tissue non-specific AP inhibitors can block adipogenesis in human preadipocytes. PMID- 16704757 TI - HLA class II DRB1 polymorphism in Argentinians undergoing chronic Trypanosoma cruzi infection. AB - DNA typing of human lymphocyte antigen (HLA)-dicloro-1-[beta]-D-ribofuranosyl benzimidazole 1 (DRB1) alleles in 35 individuals serologically positive for T. cruzi and in 41 healthy controls was performed. DRB1*0409 allele was significantly more prevalent in seropositive individuals, with a trend being also observed for the DRB1*0701 and DRB1*1503 alleles. Although statistically insignificant, the latter was found more frequent in cases with cardiomyopathy. PMID- 16704758 TI - The LH/FSH ratio has little use in diagnosing polycystic ovarian syndrome. AB - BACKGROUND: The luteinizing hormone/follicle stimulating hormone (LH/FSH) ratio is often requested to help diagnose polycystic ovarian syndrome (PCOS) despite a recent consensus recommending against its use. This study aimed to compare the variability of the LH/FSH ratio in PCOS with that of normal menstruating women over a full cycle in order to establish the diagnostic utility, or otherwise, of the test. METHODS: Twelve women with PCOS and 11 matched controls had blood collected at four-day intervals on 10 consecutive occasions over a complete menstrual cycle. RESULTS: The median LH/FSH ratio for individual subjects did not differ significantly between the PCOS and the non-affected group (1.6 versus 1.2, P = 0.14). Only 7.6% of samples from PCOS patients had an LH/FSH ratio above three, compared with 15.6% of samples from normal subjects. CONCLUSION: This study confirms that measurement of the LH/FSH ratio is of limited use in the diagnosis of PCOS. PMID- 16704759 TI - The effect of serum creatinine method choice on estimated glomerular filtration rate determined by the abbreviated MDRD formula. AB - BACKGROUND: The aim of this study was to measure the variability in estimated glomerular filtration rate (eGFR) calculated by laboratories in Northern Ireland where creatinine assays other than the Beckman CX3 assay used to derive the Modification of Diet in Renal Disease (MDRD) are utilized. METHODS: Fifty patient samples were analysed for serum creatinine by kinetic Jaffe assays on the Roche modular P-800 (compensated assay), the Beckman LX20 and the Abbott Aeroset analysers. RESULTS: The median (interquartile range) eGFR calculated by the abbreviated MDRD equation using the creatinine results obtained by each method were 45.4 (31.7-66.6), 49.2 (35.4-78.1) and 50.0 (35.1-71.2) mL/min/1.73 m2 for Beckman, Roche (compensated) and Abbott assays, respectively. Following mathematical alignment of creatinine methods to the Beckman method the median (interquartile range) eGFR for the Roche and Abbott methods were 44.0 (32.0-67.6) and 45.6 (31.2-64.8) mL/min/1.73 m2, respectively. CONCLUSION: The regression equations published in this study can be used to align creatinine methods. However, method-related biases in eGFR of up to 10% are minimal in the context of the magnitude of variance reflected in the 90% confidence intervals. PMID- 16704760 TI - Effect of plantain on therapeutic drug monitoring of digoxin and thirteen other common drugs. AB - INTRODUCTION: Plantain, a herbal remedy, has been reported to interfere with therapeutic drug monitoring of digoxin. We evaluated three commercially available plantain products for potential interference with therapeutic drug monitoring of digoxin and 13 other common drugs. METHOD: Dry content of plantain capsule or plantain leaf was extracted with either methanol or ethanol:water (60:40 by volume), added to drug-free serum and apparent digoxin was measured by both fluorescence polarization immunoassay and microparticle enzyme immunoassay. Using immunoassays, we also measured apparent concentrations of 13 other drugs (tobramycin, procainamide, tricyclic antidepressants, quinidine, carbamazepine, phenytoin, theophylline, valproic acid, amikacin, gentamycin, phenobarbital, salicylate and acetaminophen [paracetamol]) due to the presence of plantain. In separate experiments, a serum pool prepared from patients receiving digoxin was further supplemented with plantain and observed digoxin values were compared with original digoxin concentration. The presence of any cardiac glycoside in plantain was also investigated using thin layer chromatography (TLC). RESULTS: We observed no apparent digoxin in the presence of plantain in serum. Moreover, when aliquots of digoxin serum pool were supplemented with various amounts of plantain, the observed digoxin concentrations in the presence of plantain compared well with original digoxin concentration. TLC analysis did not show the presence of either digoxin or digitoxin in plantain products studied. Moreover, plantain did not affect immunoassay results of the 13 other drugs studied. CONCLUSIONS: The plantain products studied did not interfere with therapeutic drug monitoring of digoxin as well as 13 other commonly monitored drugs. PMID- 16704761 TI - Potentially clinically toxic concentrations of whole blood manganese in a patient fed enterally with a high tea consumption. AB - This report describes a 37-year-old female patient who after seven years on intermittent overnight enteral feeding supplementation was noted to have an increased whole blood manganese concentration. Manganese toxicity is well documented after pathological absorption through inhalation via the lungs, or after intravenous administration to patients on long-term total parenteral nutrition. A dietary history revealed high tea consumption. The association between high blood manganese concentrations and enteral/oral nutrition does not appear to have previously been described. PMID- 16704762 TI - Exonic deletions as a cause of erythropoietic protoporphyria. AB - Erythropoietic protoporphyria (EPP) is an inherited disorder that results from partial deficiency of ferrochelatase (FECH), the terminal enzyme of haem biosynthesis. Current methods that examine the exons and their flanking regions of the FECH gene fail to identify mutations in about one in seven of families with EPP. The presence in some families of intragenic deletions that are not identifiable by current methods for sequencing the FECH gene may partly explain the low sensitivity of mutation detection in EPP. Here we describe the identification by gene dosage analysis of a deletion of exons 3 and 4 in a family with EPP in whom a mutation had not been found by sequencing-based methods. PMID- 16704763 TI - Clinical expression of C282Y homozygous HFE haemochromatosis at 14 years of age. AB - A 14-year-old boy who presented with debilitating lethargy was shown to have an elevated serum ferritin of 572 microg/L and a C282Y homozygous HFE genotype. Liver iron concentration was measured non-invasively by magnetic resonance imaging, which revealed a liver iron concentration of 59 micromol/g dry weight (children's reference range < 14). The early phenotypic expression was further investigated by screening genomic DNA for the presence of co-inherited mutations in genes responsible for non-HFE haemochromatosis. Coding regions and splice sites in genes encoding hepcidin and haemojuvelin were sequenced and previously described mutations in ferroportin 1 and transferrin receptor 2 genes were screened. Although no mutations were found, the most likely cause for the early expression is the presence of novel mutations or gene(s). PMID- 16704764 TI - A case of low cortisol-binding globulin: use of plasma free cortisol in interpretation of hypothalamic-pituitary-adrenal axis tests. AB - We investigated a patient with suspected hypopituitarism who showed subnormal cortisol responses to stimulation tests with adrenocorticotrophic hormone (ACTH) and hypoglycaemia, but normal ACTH and 11-deoxycortisol responses in the metyrapone test. Cortisol-binding globulin (CBG) was measured by enzyme-linked immunosorbent assay (ELISA), and was used to calculate plasma free cortisol and free cortisol index. The patient had a low plasma CBG concentration. Reinterpreted in terms of free cortisol and free cortisol index, his responses to ACTH were normal. We conclude that despite subnormal total cortisol responses to ACTH and hypoglycaemia, the patient had a normal hypothalamic-pituitary-adrenal (HPA) axis. Measurement of CBG concentration and calculation of free cortisol or free cortisol index can help avoid false interpretations of dynamic tests of the HPA axis based on plasma total cortisol. PMID- 16704765 TI - NT-proBNP and exercise. PMID- 16704771 TI - Possibilities for relapsing fever reemergence. AB - Relapsing fever Borrelia infections have attracted little attention in recent years; however, where endemic, these infections still result in considerable illness and death. Despite the marked antimicrobial drug susceptibility of these organisms, therapy is often delayed through lack of clinical suspicion. With increasing travel, infections may be imported, through exotic relapsing fever infection or through resurgence of infected disease vectors. Although louseborne relapsing fever is now geographically limited, it was once of global importance. The possibility for reemergence was recently highlighted by the probable reemergence of louseborne relapsing fever in homeless persons from France. Host limitations enforced through louseborne transmission are less applicable for the tickborne forms of relapsing fever. Although the latter have reduced potential for epidemic spread, they have the ability to infect diverse hosts, thus establishing reservoirs of infection and presenting greater challenges for their control. PMID- 16704772 TI - Cost-effectiveness of West Nile virus vaccination. AB - West Nile virus (WNV) was first detected in the Western Hemisphere in 1999 in New York City. From 1999 through 2004, >16,600 cases of WNV-related illnesses were reported in the United States, of which >7,000 were neuroinvasive disease and >600 were fatal. Several approaches are under way to develop a human vaccine. Through simulations and sensitivity analysis that incorporated uncertainties regarding future transmission patterns of WNV and costs of health outcomes, we estimated that the range of values for the cost per case of WNV illness prevented by vaccination was US 20,000 dollars-59,000 dollars(mean 36,000 dollars). Cost effectiveness was most sensitive to changes in the risk for infection, probability of symptomatic illness, and vaccination cost. Analysis indicated that universal vaccination against WNV disease would be unlikely to result in societal monetary savings unless disease incidence increases substantially over what has been seen in the past 6 years. PMID- 16704773 TI - Web-based surveillance and global Salmonella distribution, 2000-2002. AB - Salmonellae are a common cause of foodborne disease worldwide. The World Health Organization (WHO) supports international foodborne disease surveillance through WHO Global Salm-Surv and other activities. WHO Global Salm-Surv members annually report the 15 most frequently isolated Salmonella serotypes to a Web-based country databank. We describe the global distribution of reported Salmonella serotypes from human and nonhuman sources from 2000 to 2002. Among human isolates, S. Enteritidis was the most common serotype, accounting for 65% of all isolates. Among nonhuman isolates, although no serotype predominated, Salmonella enterica serovar Typhimurium was reported most frequently. Several serotypes were reported from only 1 region of the world. The WHO Global Salm-Surv country databank is a valuable public health resource; it is a publicly accessible, Web based tool that can be used by health professionals to explore hypotheses related to the sources and distribution of salmonellae worldwide. PMID- 16704774 TI - Bartonella spp. in pets and effect on human health. AB - Among the many mammals infected with Bartonella spp., pets represent a large reservoir for human infection because most Bartonella spp. infecting them are zoonotic. Cats are the main reservoir for Bartonella henselae, B. clarridgeiae, and B. koehlerae. Dogs can be infected with B. vinsonii subsp. berkhoffii, B. henselae, B. clarridgeiae, B. washoensis, B. elizabethae, and B. quintana. The role of dogs as an important reservoir of Bartonella spp. is less clear than for cats because domestic dogs are more likely to be accidental hosts, at least in nontropical regions. Nevertheless, dogs are excellent sentinels for human infections because a similar disease spectrum develops in dogs. Transmission of B. henselae by cat fleas is better understood, although new potential vectors (ticks and biting flies) have been identified. We review current knowledge on the etiologic agents, clinical features, and epidemiologic characteristics of these emerging zoonoses. PMID- 16704775 TI - West Nile virus infections projected from blood donor screening data, United States, 2003. AB - National blood donor screening for West Nile virus (WNV) RNA using minipool nucleic acid amplification testing (MP-NAT) was implemented in the United States in July 2003. We compiled national NAT yield data and performed WNV immunoglobulin M (IgM) testing in 1 WNV-epidemic region (North Dakota). State specific MP-NAT yield, antibody seroprevalence, and the average time RNA is detectable by MP-NAT were used to estimate incident infections in 2003. WNV donor screening yielded 944 confirmed viremic donors. MP-NAT yield peaked in August with >0.5% of donations positive for WNV RNA in 4 states. Peak IgM seroprevalence for North Dakota was 5.2% in late September. The average time viremia is detectable by MP-NAT was 6.9 days (95% confidence interval [CI] 3.0-10.7). An estimated 735,000 (95% CI 322,000-1,147,000) infections occurred in 2003, with 256 (95% CI 112-401) infections per neuroinvasive case. In addition to preventing transfusion-transmitted WNV infection, donor screening can serve as a tool to monitor seasonal incidence in the general population. PMID- 16704776 TI - Aspergillus ustus infections among transplant recipients. AB - Aspergillus ustus is a mold that rarely infects humans; only 15 systemic cases have been reported. We report the first outbreak of invasive infection caused by A. ustus among hematopoietic stem cell transplant (HSCT) recipients. Six patients with infections were identified; 3 infections each occurred in both 2001 and 2003. Molecular typing by using randomly amplified polymorphic DNA (RAPD) and antifungal drug susceptibility testing were performed on clinical and environmental isolates recovered from our hospital from 1999 to 2003. The highest overall attack rate in HSCT patients was 1.6%. The overall death rate was 50%, and death occurred within 8 days after diagnostic culture collection. Clinical isolates exhibited decreased susceptibility to antifungal drugs, especially azoles. RAPD and phylogenetic analysis showed genetic similarity between isolates from different patients. Based on the clustering of cases in space and time and molecular data, common-source acquisition of this unusual drug-resistant species is possible. PMID- 16704777 TI - Clostridium difficile infection in patients discharged from US short-stay hospitals, 1996-2003. AB - US hospital discharges for which Clostridium difficile-associated disease (CDAD) was listed as any diagnosis doubled from 82,000 (95% confidence interval [CI] 71,000-94,000) or 31/100,000 population in 1996 to 178,000 (95% CI 151,000 205,000) or 61/100,000 in 2003; this increase was significant between 2000 and 2003 (slope of linear trend 9.48; 95% CI 6.16-12.80, p = 0.01). The overall rate during this period was severalfold higher in persons >65 years of age (228/100,000) than in the age group with the next highest rate, 45-64 years (40/100,000; p < or = 0.001). CDAD appears to be increasing rapidly in the United States and is disproportionately affecting older persons. Clinicians should be aware of the increasing risk for CDAD and make efforts to control transmission of C. difficile and prevent disease. PMID- 16704778 TI - Medication sales and syndromic surveillance, France. AB - Although syndromic surveillance systems using nonclinical data have been implemented in the United States, the approach has yet to be tested in France. We present the results of the first model based on drug sales that detects the onset of influenza season and forecasts its trend. Using weekly lagged sales of a selected set of medications, we forecast influenzalike illness (ILI) incidence at the national and regional level for 3 epidemic seasons (2000-01, 2001-02, and 2002-03) and validate the model with real-time updating on the fourth (2003-04). For national forecasts 1-3 weeks ahead, the correlation between observed ILI incidence and forecast was 0.85-0.96, an improvement over the current surveillance method in France. Our findings indicate that drug sales are a useful additional tool to syndromic surveillance, a complementary and independent source of information, and a potential improvement for early warning systems for both epidemic and pandemic planning. PMID- 16704779 TI - Personal hygiene and methicillin-resistant Staphylococcus aureus infection. AB - Methicillin-resistant Staphylococcus aureus (MRSA) infections outside the healthcare setting are an increasing concern. We conducted a case-control study to investigate an MRSA outbreak during 2002-2003 in a Missouri prison and focused on hygiene factors. Information on sociodemographic characteristics, medical history, and hygiene practices of study participants was collected by interview and medical record review. Logistic regression was used to evaluate MRSA infection in relation to hygiene factors individually and as a composite hygiene score; potential confounding factors were controlled. Selected MRSA isolates were analyzed by pulsed-field gel electrophoresis (PFGE). MRSA infection was significantly associated with a low composite hygiene score. Transmission among prison inmates appeared to be responsible for this outbreak. PFGE analysis showed that isolates were indistinguishable and associated with community-onset MRSA infections in other US prisons. Improving hygiene practices and environmental conditions may help prevent and interrupt future MRSA outbreaks in prison settings. PMID- 16704780 TI - Rickettsia prowazekii and real-time polymerase chain reaction. AB - Rickettsia prowazekii is the causative agent of epidemic typhus and a potential bioterrorism agent. Sensitive and specific rapid assays are needed to complement existing methods of detecting this organism. We developed a real-time quantitative polymerase chain reaction assay by using a species-specific probe targeting the gltA gene. This assay, which was rapid, specific for R. prowazekii only, and sensitive (cutoff detection of 1 to 5 copies per sample), detected and directly identified R. prowazekii in blood of 12 experimentally infected mice sampled at day 3 and 6 postinfection or in naturally or experimentally infected lice. Because our assay is highly standardized and easily adaptable, it could improve epidemic typhus surveillance in public health programs, especially for countries with underdiagnosed or unrecognized human cases. PMID- 16704781 TI - Serosurvey on household contacts of Marburg hemorrhagic fever patients. AB - The first major outbreak of Marburg hemorrhagic fever (MHF) outside a laboratory environment occurred in the subdistrict of Watsa, Democratic Republic of Congo, from October 1998 to August 2000. We performed a serosurvey of household contacts of MHF patients to identify undetected cases, ascertain the frequency of asymptomatic Marburg infection, and estimate secondary attack risk and postintervention reproduction number. Contacts were interviewed about their exposure and symptoms consistent with MHF. Blood samples were tested for anti Marburg immunoglobulin G (IgG). One hundred twenty-one (51%) of 237 identified contacts participated; 72 (60%) were not known to the health authorities. Two participating contacts were seropositive and reported becoming ill after the contact; no serologic evidence for asymptomatic or mild Marburg infection was found. The secondary attack risk was 21%; the postintervention reproduction number was 0.9, consistent with an outbreak sustained by repeated primary transmission, rather than large-scale secondary transmission. PMID- 16704782 TI - Canine visceral leishmaniasis, United States and Canada, 2000-2003. AB - Visceral leishmaniasis, caused by protozoa of the genus Leishmania donovani complex, is a vectorborne zoonotic infection that infects humans, dogs, and other mammals. In 2000, this infection was implicated as causing high rates of illness and death among foxhounds in a kennel in New York. A serosurvey of >12,000 foxhounds and other canids and 185 persons in 35 states and 4 Canadian provinces was performed to determine geographic extent, prevalence, host range, and modes of transmission within foxhounds, other dogs, and wild canids and to assess possible infections in humans. Foxhounds infected with Leishmania spp. were found in 18 states and 2 Canadian provinces. No evidence of infection was found in humans. The infection in North America appears to be widespread in foxhounds and limited to dog-to-dog mechanisms of transmission; however, if the organism becomes adapted for vector transmission by indigenous phlebotomines, the probability of human exposure will be greatly increased. PMID- 16704783 TI - Chemoprophylaxis and malaria death rates. AB - To determine the effect of chemoprophylaxis on the case-fatality rate of malaria, we analyzed all cases of Plasmodium falciparum malaria in nonimmune persons reported from 1993 to 2004 in Germany. In univariate and multivariate logistic regression analysis, we determined the effect of age, sex, chemoprophylaxis, chemoprophylactic regimen, compliance for chemoprophylactic regimen, exposure prophylaxis, country of infection, and year of reporting on the outcome. Of 3,935 case-patients, 116 (3%) died of malaria. Univariate analysis showed significant associations with death for chemoprophylaxis with chloroquine plus proguanil compared to no chemoprophylaxis. The multivariate model showed that patients who had taken chemoprophylaxis were less likely to die compared to those who had not taken chemoprophylaxis, adjusted for patient age and reporting year. The study demonstrated that chemoprophylaxis significantly reduced fatality rates among nonimmune malaria patients and supports the importance of existing guidelines for malaria prevention. PMID- 16704784 TI - Self-medication with antimicrobial drugs in Europe. AB - We surveyed the populations of 19 European countries to compare the prevalence of antimicrobial drug self-medication in the previous 12 months and intended self medication and storage and to identify the associated demographic characteristics. By using a multistage sampling design, 1,000-3,000 adults in each country were randomly selected. The prevalence of actual self-medication varied from 1 to 210 per 1,000 and intended self-medication from 73 to 449 per 1,000; both rates were high in eastern and southern Europe and low in northern and western Europe. The most common reasons for self-medication were throat symptoms (e.g., dry, inflamed, red, or sore throat, inflamed tonsils, tonsil pain). The main medication sources were pharmacies and medication leftover from previous prescriptions. Younger age, higher education, and presence of a chronic disease were associated with higher rates of self-medication. Attempts to reduce inappropriate self-medication should target prescribers, pharmacists, and the general public. PMID- 16704785 TI - Pneumonic plague cluster, Uganda, 2004. AB - The public and clinicians have long-held beliefs that pneumonic plague is highly contagious; inappropriate alarm and panic have occurred during outbreaks. We investigated communicability in a naturally occurring pneumonic plague cluster. We defined a probable pneumonic plague case as an acute-onset respiratory illness with bloody sputum during December 2004 in Kango Subcounty, Uganda. A definite case was a probable case with laboratory evidence of Yersinia pestis infection. The cluster (1 definite and 3 probable cases) consisted of 2 concurrent index patient-caregiver pairs. Direct fluorescent antibody microscopy and polymerase chain reaction testing on the only surviving patient's sputum verified plague infection. Both index patients transmitted pneumonic plague to only 1 caregiver each, despite 23 additional untreated close contacts (attack rate 8%). Person-to person transmission was compatible with transmission by respiratory droplets, rather than aerosols, and only a few close contacts, all within droplet range, became ill. PMID- 16704786 TI - Host feeding patterns of Culex mosquitoes and West Nile virus transmission, northeastern United States. AB - To evaluate the role of Culex mosquitoes as enzootic and epidemic vectors for WNV, we identified the source of vertebrate blood by polymerase chain reaction amplification and sequencing portions of the cytochrome b gene of mitochondrial DNA. All Cx. restuans and 93% of Cx. pipiens acquired blood from avian hosts; Cx. salinarius fed frequently on both mammals (53%) and birds (36%). Mixed-blood meals were detected in 11% and 4% of Cx. salinarius and Cx. pipiens, respectively. American robin was the most common source of vertebrate blood for Cx. pipiens (38%) and Cx. restuans (37%). American crow represented <1% of the blood meals in Cx. pipiens and none in Cx. restuans. Human-derived blood meals were identified from 2 Cx. salinarius and 1 Cx. pipiens. Results suggest that Cx. salinarius is an important bridge vector to humans, while Cx. pipiens and Cx. restuans are more efficient enzootic vectors in the northeastern United States. PMID- 16704788 TI - "Candidatus Rickettsia kellyi," India. AB - We report the first laboratory-confirmed human infection due to a new rickettsial genotype in India, "Candidatus Rickettsia kellyi," in a 1-year-old boy with fever and maculopapular rash. The diagnosis was made by serologic testing, polymerase chain reaction detection, and immunohistochemical testing of the organism from a skin biopsy specimen. PMID- 16704789 TI - Lyssavirus surveillance in bats, Bangladesh. AB - Lyssavirus surveillance in bats was performed in Bangladesh during 2003 and 2004. No virus isolates were obtained. Three serum samples (all from Pteropus giganteus, n = 127) of 288 total serum samples, obtained from bats in 9 different taxa, neutralized lyssaviruses Aravan and Khujand. The infection occurs in bats in Bangladesh, but virus prevalence appears low. PMID- 16704787 TI - Identifying and quantifying genotypes in polyclonal infections due to single species. AB - Simultaneous infection with multiple pathogens of the same species occurs with HIV, hepatitis C, Epstein-Barr virus, dengue, tuberculosis, and malaria. However, available methods do not distinguish among or quantify pathogen genotypes in individual patients; they also cannot test for novel insertions and deletions in genetically modified organisms. The strategy reported here accomplishes these goals with real-time polymerase chain reaction (PCR) and capillary electrophoresis. Real-time PCR with allotype-specific primers defines the allotypes (strains) present and the intensity of infection (copy number). Capillary electrophoresis defines the number of genotypes within each allotype and the intensity of infection by genotype. This strategy can be used to study the epidemiology of emerging infectious diseases with simultaneous infection by multiple genotypes, as demonstrated here with malaria. It also permits testing for insertions or deletions in genetically modified organisms that may be used for bioterrorism. PMID- 16704790 TI - Molecular analysis of fluoroquinolone-resistant Salmonella Paratyphi A isolate, India. AB - Salmonella enterica serovar Paratyphi A is increasingly a cause of enteric fever. Sequence analysis of an Indian isolate showed a unique strain with high-level resistance to ciprofloxacin associated with double mutations in the DNA gyrase subunit gyrA (Ser83-->Phe and Asp87-->Gly) and a mutation in topoisomerase IV subunit parC (Ser80-->Arg). PMID- 16704791 TI - Canine coronavirus highly pathogenic for dogs. AB - Canine coronavirus (CCoV) is usually responsible for mild, self-limiting infections restricted to the enteric tract. We report an outbreak of fatal disease in puppies caused by a pathogenic variant of CCoV that was isolated from organs with severe lesions. PMID- 16704792 TI - Acute hemorrhagic conjunctivitis and coxsackievirus A24v, Rio de Janeiro, Brazil, 2004. AB - An outbreak of acute hemorrhagic conjunctivitis (AHC) occurred in Rio de Janeiro in 2004. Coxsackievirus A24v (CA24v) was identified as the etiologic agent, and partial sequences from the VP1 gene show that the isolates are closely related to CA24v viruses that previously caused AHC epidemics in South Korea and French Guiana. PMID- 16704793 TI - Pneumonia and new methicillin-resistant Staphylococcus aureus clone. AB - Necrotizing pneumonia caused by Staphylococcus aureus strains carrying the Panton Valentin leukocidin gene is a newly described disease entity. We report a new fatal case of necrotizing pneumonia. An S. aureus strain with an agr1 allele and of a new sequence type 377 was recovered, representing a new, emerging, community acquired methicillin-resistant clone. PMID- 16704794 TI - Canine leptospirosis, United States, 2002-2004. AB - The proportion of positive Leptospira microscopic agglutination tests for 23,005 dogs significantly increased from 2002 to 2004 (p < 0.002) regardless of the positive cutoff titer used and was highest (p < 0.05) for serovars Autumnalis and Grippotyphosa. The strongest positive serologic correlation (r = 0.72) was between serovars Autumnalis and Pomona. PMID- 16704795 TI - Lagos bat virus, South Africa. AB - Three more isolates of Lagos bat virus were recently recovered from fruit bats in South Africa after an apparent absence of this virus for 13 years. The sporadic occurrence of cases is likely due to inadequate surveillance programs for lyssavirus infections among bat populations in Africa. PMID- 16704796 TI - Aquariums as reservoirs for multidrug-resistant Salmonella Paratyphi B. AB - Multidrug-resistant Salmonella enterica serovar Paratyphi B dT+ isolates from patients with gastroenteritis were identical with isolates from their home aquariums. Matched isolates had identical phage types, XbaI and IS200 profiles, and Salmonella genomic island 1 (SGI1). Ornamental fish tanks are reservoirs for SGI1-containing S. Paratyphi B dT+. PMID- 16704797 TI - Protease-resistant prion protein in lymphoreticular tumors of variant Creutzfeldt Jakob disease mice. AB - We report protease-resistant prion protein (PrPres) in spontaneous lymphoreticular tumors of mice infected with the agent of variant Creutzfeldt Jakob disease (vCJD). PrPres may accumulate in lymphoreticular system tumors of asymptomatic persons with vCJD. The statistical power of estimates of vCJD prevalence might be increased by expanding screening to include samples of lymphoreticular neoplasms. PMID- 16704798 TI - West Nile Virus-associated flaccid paralysis outcome. AB - We report 1-year follow-up data from a longitudinal prospective cohort study of patients with West Nile virus-associated paralysis. As in the 4-month follow-up, a variety of recovery patterns were observed, but persistent weakness was frequent. Respiratory involvement was associated with considerable illness and death. PMID- 16704799 TI - Negligible risk for epidemics after geophysical disasters. AB - After geophysical disasters (i.e., earthquakes, volcanic eruptions, tsunamis), media reports almost always stress the risk for epidemics; whether this risk is genuine has been debated. We analyzed the medical literature and data from humanitarian agencies and the World Health Organization from 1985 to 2004. Of >600 geophysical disasters recorded, we found only 3 reported outbreaks related to these disasters: 1 of measles after the eruption of Pinatubo in Philippines, 1 of coccidioidomycosis after an earthquake in California, and 1 of Plasmodium vivax malaria in Costa Rica related to an earthquake and heavy rainfall. Even though the humanitarian response may play a role in preventing epidemics, our results lend support to the epidemiologic evidence that short-term risk for epidemics after a geophysical disaster is very low. PMID- 16704800 TI - Potential arbovirus emergence and implications for the United Kingdom. AB - Arboviruses have evolved a number of strategies to survive environmental challenges. This review examines the factors that may determine arbovirus emergence, provides examples of arboviruses that have emerged into new habitats, reviews the arbovirus situation in western Europe in detail, discusses potential arthropod vectors, and attempts to predict the risk for arbovirus emergence in the United Kingdom. We conclude that climate change is probably the most important requirement for the emergence of arthropodborne diseases such as dengue fever, yellow fever, Rift Valley fever, Japanese encephalitis, Crimean-Congo hemorrhagic fever, bluetongue, and African horse sickness in the United Kingdom. While other arboviruses, such as West Nile virus, Sindbis virus, Tahyna virus, and Louping ill virus, apparently circulate in the United Kingdom, they do not appear to present an imminent threat to humans or animals. PMID- 16704801 TI - Confronting zoonoses, linking human and veterinary medicine. AB - Many of the emerging infectious diseases, including those caused by bioterrorist agents, are zoonoses. Since zoonoses can infect both animals and humans, the medical and veterinary communities should work closely together in clinical, public health, and research settings. In the clinical setting, input from both professions would improve assessments of the risk-benefit ratios of pet ownership, particularly for pet owners who are immunocompromised. In public health, human and animal disease surveillance systems are important in tracking and controlling zoonoses such as avian influenza virus, West Nile virus, and foodborne pathogens. Comparative medicine is the study of disease processes across species, including humans. Physician and veterinarian comparative medicine research teams should be promoted and encouraged to study zoonotic agent-host interactions. These efforts would increase our understanding of how zoonoses expand their host range and would, ultimately, improve prevention and control strategies. PMID- 16704802 TI - Human influenza surveillance: the demand to expand. AB - The World Health Organization Influenza Program is one of the best developed and longest running infectious disease surveillance systems that exists. It maintains a worldwide watch of influenza's evolution to assist delivery of appropriately formulated vaccines in time to blunt seasonal epidemics and unpredictable pandemics. Despite the program's success, however, much more is possible with today's advanced technologies. This article summarizes ongoing human influenza surveillance activities worldwide. It shows that the technology to establish a high-throughput laboratory network that can process and test influenza viruses more quickly and more accurately is available. It also emphasizes the practical public health and scientific applications of such a network. PMID- 16704803 TI - Prospects for universal influenza virus vaccine. AB - The current vaccination strategy against influenza A and B viruses is vulnerable to the unanticipated emergence of epidemic strains that are poorly matched by the vaccine. A vaccine that is less sensitive to the antigenic evolution of the virus would be a major improvement. The general feasibility of this goal is supported by studies in animal models that show that immunologic activities directed against relatively invariant viral determinants can reduce illness and death. The most promising approaches are based on antibodies specific for the relatively conserved ectodomain of matrix protein 2 and the intersubunit region of hemagglutinin. However, additional conserved determinants for protective antibodies are likely to exist, and their identification should be encouraged. Most importantly, infection and current vaccines do not appear to effectively induce these antibodies in humans. This finding provides a powerful rationale for testing the protective activity of these relatively conserved viral components in humans. PMID- 16704804 TI - Domestic ducks and H5N1 influenza epidemic, Thailand. AB - In addition to causing 12 human deaths and 17 cases of human infection, the 2004 outbreak of H5N1 influenza virus in Thailand resulted in the death or slaughter of 60 million domestic fowl and the disruption of poultry production and trade. After domestic ducks were recognized as silent carriers of H5N1 influenza virus, government teams went into every village to cull flocks in which virus was detected; these team efforts markedly reduced H5N1 infection. Here we examine the pathobiology and epidemiology of H5N1 influenza virus in the 4 systems of duck raising used in Thailand in 2004. No influenza viruses were detected in ducks raised in "closed" houses with high biosecurity. However, H5N1 influenza virus was prevalent among ducks raised in "open" houses, free-ranging (grazing) ducks, and backyard ducks. PMID- 16704805 TI - Recently acquired Toxoplasma gondii infection, Brazil. AB - The city of Erechim, Brazil, has a 17% prevalence of ocular toxoplasmosis, and type 1 Toxoplasma gondii predominates. To examine risk factors for acute T. gondii infection in this area, we administered a questionnaire to recently infected persons (n = 131) and seronegative controls (n = 110). Eating undercooked meat; having a garden; working in the garden or yard more than once per week; eating rare meat; eating cured, dried, or smoked meat; eating frozen lamb; and being male increased risk for T. gondii infection in univariate analysis. Risk factors independently associated with acute T. gondii infection in multivariate analysis were working in the garden (odds ratio [OR] 2.35, 95% confidence interval [CI] 1.27-4.33) and eating frozen lamb (OR 2.06, 95% CI 1.15 3.67). Among women (n = 86), having had children markedly increased the risk for T. gondii infection (OR 14.94, 95% CI 3.68-60.73). PMID- 16704806 TI - Reducing Legionella colonization in water systems with monochloramine. AB - Monochloramine disinfection of municipal water supplies is associated with decreased risk for Legionnaires' disease. We conducted a 2-year, prospective, environmental study to evaluate whether converting from chlorine to monochloramine for water disinfection would decrease Legionella colonization of hot water systems. Water and biofilm samples from 53 buildings were collected for Legionella culture during 6 intervals. Prevalence ratios (PRs) comparing Legionella colonization before and after monochloramine disinfection were adjusted for water system characteristics. Legionella colonized 60% of the hot water systems before monochloramine versus 4% after conversion (PR 0.07, 95% confidence interval 0.03-0.16). The median number of colonized sites per building decreased with monochloramine disinfection. Increased prevalence of Legionella colonization was associated with water heater temperatures <50 degrees C, buildings taller than 10 stories, and interruptions in water service. Increasing use of monochloramine in water supplies throughout the United States may reduce Legionella transmission and incidence of Legionnaires' disease. PMID- 16704807 TI - Atypical enteropathogenic Escherichia coli infection and prolonged diarrhea in children. AB - Some clinical isolates of enteropathogenic Escherichia coli (EPEC) lack bundle forming pili and are termed atypical EPEC. The aim of this study was to determine if atypical EPEC are pathogens by comparing the clinical features of patients infected with atypical EPEC with those of children infected with other causative agents of diarrhea. Fecal samples obtained from children attending the Royal Children's Hospital in Melbourne for investigation of diarrhea were examined for adenovirus, rotavirus, Campylobacter spp., Salmonella spp., protozoa, and pathogenic E. coli. Clinical data were obtained by using a standardized pro forma and analyzed separately. Patients infected with atypical EPEC experienced mild, nondehydrating, and noninflammatory diarrhea that was not particularly associated with fever, vomiting, or abdominal pain. However, the duration of diarrhea in patients infected with atypical EPEC was significantly longer than that caused by the other species or where no pathogens were identified. Infection with atypical EPEC is associated with prolonged diarrhea. PMID- 16704808 TI - Epidemic spread of Lyme borreliosis, northeastern United States. AB - We examined the degree of host specialization of different strains of Borrelia burgdorferi, the tickborne pathogen that causes Lyme borreliosis in the northeastern United States. We first assessed the genetic population structures of B. burgdorferi in ticks obtained from different mammalian host species and in questing ticks sampled in a woodland ecosystem in Connecticut. By comparing the patterns found in our study with data from another cross-sectional study, we demonstrate that B. burgdorferi is a generalist microparasite and conclude that efficient cross-species transmission of B. burgdorferi is a key feature that has allowed the rapid spread of Lyme borreliosis across the northeastern United States. PMID- 16704809 TI - Emerging pediatric HIV epidemic related to migration. AB - In 2002, Canada introduced routine, mandatory HIV antibody screening for all residency applicants, including selected children. We report screening results from January 2002 to February 2005. Thirty-six pediatric HIV cases were detected (14/100,000 applicants); 94% of infected children were eligible to arrive in Canada. Thirty-two of the affected children were from Africa, and maternal infection was the main risk factor. Only 4 (11%) of the children had received antiretroviral therapy. In countries of low HIV incidence, migration-related imported infection in children may be an emerging epidemic. The early identification of HIV-infected immigrant women permits intervention to prevent mother-to-child HIV transmission. Routine HIV testing as a component of the medical examination of immigrants has national and international health policy and programmatic implications. PMID- 16704810 TI - Lineage 1 and 2 strains of encephalitic West Nile virus, central Europe. AB - Two different West Nile virus (WNV) strains caused lethal encephalitis in a flock of geese and a goshawk in southeastern Hungary in 2003 and 2004, respectively. During the outbreak in geese, 14 confirmed human cases of WNV encephalitis and meningitis were reported in the same area. Sequencing of complete genomes of both WNV strains and phylogenetic analyses showed that the goose-derived strain exhibits closest genetic relationship to strains isolated in 1998 in Israel and to the strain that emerged in 1999 in the United States. WNV derived from the goshawk showed the highest identity to WNV strains of lineage 2 isolated in central Africa. The same strain reemerged in 2005 in the same location, which suggests that the virus may have overwintered in Europe. The emergence of an exotic WNV strain in Hungary emphasizes the role of migrating birds in introducing new viruses to Europe. PMID- 16704811 TI - Human Trypanosoma cruzi infection and seropositivity in dogs, Mexico. AB - We used 5 diagnostic tests in a cross-sectional investigation of the prevalence of Trypanosoma cruzi in Tejupilco municipality, State of Mexico, Mexico. Our findings showed a substantial prevalence of immunoglobulin G (IgG) and IgM antibodies to T. cruzi in human (n = 293, IgG 2.05%, IgM 5.5%, both 7.1%) and dog (n = 114, IgG 15.8%, IgM 11.4%, both 21%) populations. We also found antibodies to T. cruzi (n = 80, IgG 10%, IgM 15%, both 17.5%) in dogs from Toluca, an area previously considered free of T. cruzi. Our data demonstrate the need for active epidemiologic surveillance programs in these regions. A direct correlation (r2 = 0.955) of seropositivity between humans and dogs suggests that seroanalysis in dogs may help identify the human prevalence of T. cruzi infection in these areas. PMID- 16704812 TI - Contrasting pediatric and adult methicillin-resistant Staphylococcus aureus isolates. AB - We performed a prospective study of all inpatient and outpatient methicillin resistant Staphylococcus aureus (MRSA) isolates identified at the University of Chicago Hospitals from November 2003 through November 2004. Differences in resistance to non-beta-lactam antimicrobial drugs were determined after stratification of the 578 MRSA isolates into 4 groups by patient age (pediatric vs. adult) and onset location (community vs. hospital). Non-beta-lactam resistance was significantly greater among the 288 adult than the 177 pediatric community-associated isolates for erythromycin (93.2 vs. 87.0%, p = 0.03), clindamycin (51.8 vs. 7.3%, p<0.001), ciprofloxacin (62.1 vs. 10.7%, p<0.001), gentamicin (11.1 vs. 1.1%, p<0.001), and tetracycline (19.9 vs. 6.4%, p<0.001). In contrast, hospital-associated MRSA isolates from children and adults had similar rates of non-beta-lactam antimicrobial drug resistance. In our region, clindamycin is an appropriate empiric therapy of community-associated MRSA infection in children but should be used with caution in adults. PMID- 16704813 TI - Identifying influenza viruses with resequencing microarrays. AB - Identification of genetic variations of influenza viruses is essential for epidemic and pandemic outbreak surveillance and determination of vaccine strain selection. In this study, we combined a random amplification strategy with high density resequencing microarray technology to demonstrate simultaneous detection and sequence-based typing of 25 geographically distributed human influenza virus strains collected in 2004 and 2005. In addition to identification, this method provided primary sequence information, which suggested that distinct lineages of influenza viruses co-circulated during the 2004-2005 season, and simultaneously identified and typed all component strains of the trivalent FluMist intranasal vaccine. The results demonstrate a novel, timely, and unbiased method for the molecular epidemiologic surveillance of influenza viruses. PMID- 16704814 TI - Animals as sentinels of bioterrorism agents. AB - We conducted a systematic review of the scientific literature from 1966 to 2005 to determine whether animals could provide early warning of a bioterrorism attack, serve as markers for ongoing exposure risk, and amplify or propagate a bioterrorism outbreak. We found evidence that, for certain bioterrorism agents, pets, wildlife, or livestock could provide early warning and that for other agents, humans would likely manifest symptoms before illness could be detected in animals. After an acute attack, active surveillance of wild or domestic animal populations could help identify many ongoing exposure risks. If certain bioterrorism agents found their way into animal populations, they could spread widely through animal-to-animal transmission and prove difficult to control. The public health infrastructure must look beyond passive surveillance of acute animal disease events to build capacity for active surveillance and intervention efforts to detect and control ongoing outbreaks of disease in domestic and wild animal populations. PMID- 16704815 TI - Economic impact of Lyme disease. AB - To assess the economic impact of Lyme disease (LD), the most common vectorborne inflammatory disease in the United States, cost data were collected in 5 counties of the Maryland Eastern Shore from 1997 to 2000. Patients were divided into 5 diagnosis groups, clinically defined early-stage LD, clinically defined late stage LD, suspected LD, tick bite, and other related complaints. From 1997 to 2000, the mean per patient direct medical cost of early-stage LD decreased from $1,609 to $464 (p<0.05), and the mean per patient direct medical cost of late stage LD decreased from $4,240 to $1,380 (p<0.05). The expected median of all costs (direct medical cost, indirect medical cost, nonmedical cost, and productivity loss), aggregated across all diagnosis groups of patients, was approximately $281 per patient. These findings will help assess the economics of current and future prevention and control efforts. PMID- 16704816 TI - 1951 influenza epidemic, England and Wales, Canada, and the United States. AB - Influenza poses a continuing public health threat in epidemic and pandemic seasons. The 1951 influenza epidemic (A/H1N1) caused an unusually high death toll in England; in particular, weekly deaths in Liverpool even surpassed those of the 1918 pandemic. We further quantified the death rate of the 1951 epidemic in 3 countries. In England and Canada, we found that excess death rates from pneumonia and influenza and all causes were substantially higher for the 1951 epidemic than for the 1957 and 1968 pandemics (by > or =50%). The age-specific pattern of deaths in 1951 was consistent with that of other interpandemic seasons; no age shift to younger age groups, reminiscent of pandemics, occurred in the death rate. In contrast to England and Canada, the 1951 epidemic was not particularly severe in the United States. Why this epidemic was so severe in some areas but not others remains unknown and highlights major gaps in our understanding of interpandemic influenza. PMID- 16704817 TI - HIV transmission in correctional facility. AB - Acute retroviral syndrome developed in an inmate in a detention center after he had intercourse with 2 HIV-infected inmates. Correctional facilities house a disproportionate number of HIV-infected persons, and most do not provide inmates with condoms. Correctional healthcare providers should be familiar with primary HIV infection and acute retroviral syndrome. PMID- 16704818 TI - Fatal human infection with Rickettsia rickettsii, Yucatan, Mexico. AB - The first fatal Rickettsia rickettsii infection was diagnosed in the southwest of Mexico. The patient had fever, erythematous rash, abdominal pain, and severe central nervous system involvement with convulsive crisis. The diagnosis of R. rickettsii infection was established by immunohistochemistry and specific polymerase chain reaction. PMID- 16704819 TI - Shrews as reservoir hosts of borna disease virus. AB - Borna disease virus (BDV) is the causative agent of severe T-cell-mediated meningoencephalitis in horses, sheep, and other animal species in central Europe. Here we report the first unequivocal detection of a BDV reservoir species, the bicolored white-toothed shrew, Crocidura leucodon, in an area in Switzerland with endemic Borna disease. PMID- 16704820 TI - Tularemia outbreak, Bulgaria, 1997-2005. AB - The 1997-2005 tularemia outbreak in Bulgaria affected 285 people. Ten strains were isolated from humans, a tick, a hare, and water. Amplified fragment length polymorphism typing of the present isolates and of the strain isolated in 1962 suggests that a new genetic variant caused the outbreak. PMID- 16704821 TI - Avian influenza H5N1 in naturally infected domestic cat. AB - We report H5N1 virus infection in a domestic cat infected by eating a pigeon carcass. The virus isolated from the pigeon and the cat showed the same cluster as the viruses obtained during the outbreak in Thailand. Since cats are common house pets, concern regarding disease transmission to humans exists. PMID- 16704822 TI - Cryptosporidiosis associated with ozonated apple cider. AB - We linked an outbreak of cryptosporidiosis to ozonated apple cider by using molecular and epidemiologic methods. Because ozonation was insufficient in preventing this outbreak, its use in rendering apple cider safe for drinking is questioned. PMID- 16704823 TI - Long-term follow-up for multidrug-resistant tuberculosis. AB - Patients treated in Peru for multidrug-resistant tuberculosis (MDR-TB) were followed-up for a median of 67 months. Among 86 patients considered cured after completion of treatment, 97% remain healthy; 1 patient relapsed. Employment increased from 34% before treatment to 71%. We observed favorable long-term outcomes among MDR-TB patients. PMID- 16704824 TI - Immune restoration disease in HIV patient. AB - We describe a severely immunosuppressed HIV-1-positive man in whom immune restoration disease associated with pulmonary infection caused by Mycobacterium microti developed after antiretroviral treatment. The diagnosis was made by using convenient spoligotyping techniques, but invasive investigations were required to exclude a tumor. PMID- 16704826 TI - Universal genotyping in tuberculosis control program, New York City, 2001-2003. AB - In 2001, New York City implemented genotyping to its tuberculosis (TB) control activities by using IS6110 restriction fragment length polymorphism (RFLP) and spoligotyping to type isolates from culture-positive TB patients. Results are used to identify previously unknown links among genotypically clustered patients, unidentified sites of transmission, and potential false-positive cultures. From 2001 to 2003, spoligotype and IS6110-based RFLP results were obtained for 90.7% of eligible and 93.7% of submitted isolates. Fifty-nine (2.4%) of 2,437 patient isolates had false-positive culture results, and 205 genotype clusters were identified, with 2-81 cases per cluster. Cluster investigations yielded 57 additional links and 17 additional sites of transmission. Four additional TB cases were identified as a result of case finding initiated through cluster investigations. Length of unnecessary treatment decreased among patients with false-positive cultures. PMID- 16704825 TI - MassTag polymerase chain reaction for differential diagnosis of viral hemorrhagic fever. AB - Viral hemorrhagic fevers are associated with high rates of illness and death. Although therapeutic options are limited, early differential diagnosis has implications for containment and may aid in clinical management. We describe a diagnostic system for rapid, multiplex polymerase chain reaction identification of 10 different causes of viral hemorrhagic fevers. PMID- 16704827 TI - Tuberculin skin testing in children. AB - In 1996, the American Academy of Pediatrics (AAP) recommended targeted tuberculin skin testing (TST) of children while discouraging routine TST of children without risk factors for tuberculosis (TB). Recent studies have provided evidence in support of the targeted TST and recommendations that favor risk assessment over universal screening with TST. While evidence for targeted TB testing exists and benefits of screening programs are clear, administrative logistics could be a greater issue. The challenge for public health and school officials is to develop a screening program that avoids stigmatization of the at-risk group. Until then, pediatric healthcare providers will continue to have a key role in identifying children at risk for latent TB infection by using the AAP-endorsed risk assessment questionnaire and should screen children with TST only when >1 risk factor is present. PMID- 16704828 TI - Tuberculosis transmission attributable to close contacts and HIV status, Malawi. AB - We conducted the first molecular study of tuberculosis (TB) to estimate the role of household contact and transmission from HIV-positive putative source contacts (PSCs) in a high HIV-prevalence area. TB patients in a long-term population-based study in Malawi were asked about past contact with TB. DNA fingerprinting was used to define clusters of cases with identical strains. Among 143 epidemiologically defined PSC-case pairs, fingerprinting confirmed transmission for 44% of household and family contacts and 18% of other contacts. Transmission was less likely to be confirmed if the PSC were HIV positive than if he or she was HIV negative (odds ratio 0.32, 95% confidence interval [CI] 0.14-0.74). Overall, epidemiologic links were found for 11% of 754 fingerprint-clustered cases. We estimate that 9%-13% of TB cases were attributable to recent transmission from identifiable close contacts and that nearly half of the TB cases arising from recent infection had acquired the infection from HIV-positive patients. PMID- 16704830 TI - Isoniazid preventive therapy and risk for resistant tuberculosis. AB - In the context of tuberculosis (TB) resurgence, isoniazid preventive therapy (IPT) is increasingly promoted, but concerns about the risk for development of isoniazid-resistant tuberculosis may hinder its widespread implementation. We conducted a systematic review of data published since 1951 to assess the effect of primary IPT on the risk for isoniazid-resistant TB. Different definitions of isoniazid resistance were used, which affected summary effect estimates; we report the most consistent results. When all 13 studies (N = 18,095 persons in isoniazid groups and N = 17,985 persons in control groups) were combined, the summary relative risk for resistance was 1.45 (95% confidence interval 0.85 2.47). Results were similar when studies of HIV-uninfected and HIV-infected persons were considered separately. Analyses were limited by small numbers and incomplete testing of isolates, but findings do not exclude an increased risk for isoniazid-resistant TB after IPT. The diagnosis of active TB should be excluded before IPT. Continued surveillance for isoniazid resistance is essential. PMID- 16704831 TI - Mycobacterium tuberculosis and rifampin resistance, United Kingdom. AB - The United Kingdom Health Protection Agency Mycobacterium Reference Unit offers a national "Fastrack" molecular service for detecting Mycobacterium tuberculosis complex (MTBC) and rifampin resistance by using the INNO-LiPA Rif.TB assay. We analyzed the service in a routine, nontrial context of 1,997 primary clinical specimens, including 658 nonrespiratory specimens. The overall adjusted concordance, sensitivity, specificity, positive predictive value, and negative predictive value for detecting MTBC were 91.2%, 85.2%, 96.2%, 95.7%, and 86.7%, respectively (unadjusted, 86.7%, 85.2%, 88.2%, 86.9%, and 86.7%), when false positive samples from patients (n = 83) with a known microbiologic diagnosis of MTBC or patients receiving current or recent antituberculous treatment were excluded. The parameters for detecting rifampin resistance were 99.1%, 95.0%, 99.6%, 92.7%, and 99.7%, respectively. The assay enabled earlier diagnosis of MTBC and rifampin resistance (15.2 days) compared with culture-based techniques (30.7 days). PMID- 16704829 TI - Beijing/W genotype Mycobacterium tuberculosis and drug resistance. AB - Beijing/W genotype Mycobacterium tuberculosis is widespread, may be increasing, and may have a predilection for drug resistance. Individual-level data on >29,000 patients from 49 studies in 35 countries were combined to assess the Beijing genotype's prevalence worldwide, trends over time and with age, and associations with drug resistance. We found 4 patterns for Beijing/W genotype tuberculosis (TB): 1) endemic, not associated with drug resistance (high level in most of East Asia, lower level in parts of the United States); 2) epidemic, associated with drug resistance (high level in Cuba, the former Soviet Union, Vietnam, and South Africa, lower level in parts of Western Europe); 3) epidemic but drug sensitive (Malawi, Argentina); and 4) very low level or absent (parts of Europe, Africa). This study confirms that Beijing/W genotype TB is an emerging pathogen in several areas and a predominant endemic strain in others; it is frequently associated with drug resistance. PMID- 16704832 TI - Multidrug-resistant tuberculosis in military recruits. AB - We conducted a tuberculosis contact investigation for a female military recruit with an unreported history of multidrug-resistant tuberculosis (MDRTB) and subsequent recurrence. Pertinent issues included identification of likely contacts from separate training phases, uncertainty on latent MDRTB infection treatment regimens and side effects, and subsequent dispersal of the contacts after exposure. PMID- 16704833 TI - Mycobacterium bovis isolates with M. tuberculosis specific characteristics. AB - Our study is the first report of exceptional Mycobacterium bovis strains that have some characteristics of M. tuberculosis. The strains were isolated from 8 patients living in Kazakhstan. While molecular markers were typical for M. bovis, growth characteristics and biochemical test results were intermediate between M. bovis and M. tuberculosis. PMID- 16704834 TI - Tuberculosis-HIV co-infection in Kiev City, Ukraine. AB - In 2004, we tested all patients with newly diagnosed tuberculosis (TB) for HIV in Kiev City. The results were compared to information from medical records of 2002, when co-infection prevalence was 6.3%. Of 968 TB patients, 98 (10.1%) were HIV infected. TB-HIV co-infection is increasing, especially in injecting drug users. PMID- 16704835 TI - Mycobacterium bovis isolates from tuberculous lesions in Chadian zebu carcasses. AB - This slaughterhouse study in Chad shows higher proportions of Mycobacterium bovis isolates among Mbororo than Arabe zebu cattle. Spoligotyping shows a homogenetic population structure for M. bovis and lack of spacer 30, as were found in neighboring Cameroon and Nigeria. This finding suggests transborder and ongoing transmission between cattle. PMID- 16704836 TI - Intact pks15/1 in non-W-Beijing Mycobacterium tuberculosis isolates. AB - To determine whether intact pks15/1 is unique to the W-Beijing family, we investigated 147 Mycobacterium tuberculosis strains with different IS6110 genotypes. Intact pks15/1 was found in 87.8% of cerebrospinal fluid and 84.9% of sputum isolates. It was found not only in W-Beijing strains (approximate, equals 97%) but also in other genotypes (38.5%-100%). PMID- 16704837 TI - Coronavirus HKU1 infection in the United States. AB - In 2005, a new human coronavirus, HCoV-HKU1, was identified in Hong Kong. We screened respiratory specimens collected from December 16, 2001, to December 15, 2002, from children <5 years of age who tested negative for respiratory syncytial virus, parainfluenza viruses, influenza virus, and adenovirus for HCoV-HKU1 by reverse transcription-polymerase chain reaction. Overall, 1,048 respiratory specimens from 851 children were tested, and 9 HCoV-HKU1-positive children (1%) were identified, 2 of whom had 2 positive specimens. Children who had HCoV-HKU1 infection had evidence of either upper or lower respiratory tract infection or both. Two patients had disease beyond the respiratory tract. HCoV-HKU1 was identified from December 2001 to February 2002. Sequence analyses suggest that a single strain was circulating. HCoV-HKU1 is therefore likely circulating in the United States and is associated with upper and lower respiratory tract disease. PMID- 16704838 TI - Shiga-toxigenic Escherichia coli O157 in agricultural fair livestock, United States. AB - Agricultural fairs exhibiting livestock are increasingly implicated in human Shiga-toxigenic Escherichia coli O157:H7 (STEC O157:H7) outbreaks. To estimate livestock STEC O157:H7 prevalence at US fairs, we collected 2,919 fecal specimens at 29 county fairs in 2 states and at 3 state fairs in 2002. Fly pools were also collected. STEC O157:H7 was isolated from livestock at 31 (96.9%) of 32 fairs, including 11.4% of 1,407 cattle, 1.2% of 1,102 swine, 3.6% of 364 sheep and goats, and 5.2% of 154 fly pools. Cattle, swine, and flies at some fairs shared indistinguishable STEC O157:H7 isolate subtypes. In 2003, a total of 689 ambient environmental samples were collected at 20 fairgrounds 10-11 months after 2002 livestock sampling while fairgrounds were livestock-free. Four beef barn environmental samples at 3 fairgrounds yielded STEC O157:H7. These data suggest that STEC O157 is common and transmissible among livestock displayed at agricultural fairs and persists in the environment after the fair. PMID- 16704839 TI - Novel swine influenza virus subtype H3N1, United States. AB - Influenza A virus infects various animal species and transmits among different hosts, especially between humans and swine. Swine may serve as a mixing vessel to create new reassortants that could infect humans. Thus, monitoring and characterizing influenza viruses in swine are important in preventing interspecies transmission. We report the emergence and characterization of a novel H3N1 subtype of swine influenza virus (SIV) in the United States. Phylogenetic analysis showed that the H3N1 SIVs may have acquired the hemagglutinin gene from an H3N2 turkey isolate, the neuraminidase gene from a human H1N1 isolate, and the remaining genes from currently circulating SIVs. The H3N1 SIVs were antigenically related to the turkey virus. Lung lesions and nasal shedding occurred in swine infected with the H3N1 SIVs, suggesting the potential to transmit among swine and to humans. Further surveillance will help determine whether this novel subtype will continue to circulate in swine populations. PMID- 16704840 TI - The Trojan Chicken study, Minnesota. AB - We conducted a study in the summer of 2004 at county fairs in the Midwest to investigate the role poultry exhibits have in spreading avian pathogens to humans. A nearly invisible powder (pathogen surrogate) that fluoresces under UV light was surreptitiously sprinkled each day on 1 show bird at each of 2 fairs. A UV light box was used to daily examine the hands of 94 poultry-exhibit participants (blinded regarding UV box results) for up to 4 days during the poultry shows. Enrollment and end-of-study questionnaires collected data on pathogen risk factors. Eight (8.5%) of 94 participants had evidence of fluorescent powder contamination (95% confidence interval 2.76%-14.26%). This contamination and infrequent handwashing practices suggest that county fairs are a possible venue for animal-to-human pathogen transmission. PMID- 16704841 TI - Aedes aegypti larval indices and risk for dengue epidemics. AB - We assessed in a case-control study the test-validity of Aedes larval indices for the 2000 Havana outbreak. "Cases" were blocks where a dengue fever patient lived during the outbreak. "Controls" were randomly sampled blocks. Before, during, and after the epidemic, we calculated Breteau index (BI) and house index at the area, neighborhood, and block level. We constructed receiver operating characteristic (ROC) curves to determine their performance as predictors of dengue transmission. We observed a pronounced effect of the level of measurement. The BI(max) (maximum block BI in a radius of 100 m) at 2-month intervals had an area under the ROC curve of 71%. At a cutoff of 4.0, it significantly (odds ratio 6.00, p<0.05) predicted transmission with 78% sensitivity and 63% specificity. Analysis of BI at the local level, with human-defined boundaries, could be introduced in control programs to identify neighborhoods at high risk for dengue transmission. PMID- 16704842 TI - Enterobacter cloacae outbreak and emergence of quinolone resistance gene in Dutch hospital. AB - An outbreak of Enterobacter cloacae infections with variable susceptibility to fluoroquinolones occurred in the University Medical Center Utrecht in the Netherlands in 2002. Our investigation showed that a qnrA1 gene was present in 78 (94%) of 83 outbreak isolates and that a qnrA1-encoding plasmid transferred to other strains of the same species and other species. The earliest isolate carrying this same plasmid was isolated in 1999. qnrA1 was located in a complex integron consisting of the intI1, aadB, qacEDelta1, sul1, orf513, qnrA1, ampR, qacEDelta1, and sul1 genes that were not described previously. On the same plasmid, 2 other class 1 integrons were present. One was a new integron associated with the bla(CTX-M-9) extended-spectrum beta-lactamase. PMID- 16704843 TI - Spatial analysis of sleeping sickness, southeastern Uganda, 1970-2003. AB - Sleeping sickness re-emerged in southeastern Uganda in the 1970s and remains a public health problem. It has continued to spread north into new districts, and gaps remain in the understanding of the causes of its spread and distribution. We report the distribution and magnitude of sleeping sickness in southeastern Uganda from 1970 to 2003. Data were collected from records of the Ugandan Ministry of Health, individual sleeping sickness treatment centers, and interviews with public health officials. Data were used to develop incidence maps over time, conduct space-time cluster detection analyses, and develop a velocity vector map to visualize spread of sleeping sickness over time in southeastern Uganda. Results show rapid propagation of sleeping sickness from its epicenter in southern Iganga District and its spread north into new districts and foci. PMID- 16704844 TI - Mycobacterium intermedium granulomatous dermatitis from hot tub exposure. AB - Nontuberculous mycobacteria, which are widespread in the environment, frequently cause opportunistic infections in immunocompromised patients. We report the first case of a patient with chronic granulomatous dermatitis caused by a rarely described organism, Mycobacterium intermedium. The infection was associated with exposure in a home hot tub. PMID- 16704845 TI - Molecular characterization of rotavirus gastroenteritis strains, Iraqi Kurdistan. AB - Of 260 children with acute diarrhea in Erbil, Iraqi Kurdistan, 96 (37%) were infected with rotavirus. Reverse transcription-polymerase chain reaction identified G1, G4, G2, G9, P[8], P[6], and P[4] as the most common genotypes. Eight G/P combinations were found, but P[8]G1 and P[4]G2 accounted for >50% of the strains. PMID- 16704846 TI - Clostridium difficile ribotype 027, toxinotype III, the Netherlands. AB - Outbreaks due to Clostridium difficile polymerase chain reaction (PCR) ribotype 027, toxinotype III, were detected in 7 hospitals in the Netherlands from April 2005 to February 2006. One hospital experienced at the same time a second outbreak due to a toxin A-negative C. difficile PCR ribotype 017 toxinotype VIII strain. The outbreaks are difficult to control. PMID- 16704847 TI - Costs of surgical site infections that appear after hospital discharge. AB - Data were collected from surgical patients in the hospital and on 4 occasions postdischarge. The incidence of postdischarge surgical site infection was 8.46%. Strong evidence showed that these infections caused minor additional costs, which contradicts existing literature. We discuss why previous studies might have overstated costs. PMID- 16704848 TI - Historical Lassa fever reports and 30-year clinical update. AB - Five cases of Lassa fever have been imported from West Africa to the United States since 1969. We report symptoms of the patient with the second imported case and the symptoms and long-term follow-up on the patient with the third case. Vertigo in this patient has persisted for 30 years. PMID- 16704850 TI - Rickettsia felis in fleas, Western Australia. AB - This study is the first confirmation of Rickettsia felis in Australia. The organism was identified from 4 species of fleas obtained from dogs and cats in Western Australia, by using polymerase chain reaction amplification and DNA sequencing of the citrate synthase and outer membrane protein A genes. PMID- 16704849 TI - Hantavirus in African wood mouse, Guinea. AB - Hantaviruses are rodent-borne, emerging viruses that cause life-threatening human diseases in Eurasia and the Americas. We detected hantavirus genome sequences in an African wood mouse (Hylomyscus simus) captured in Sangassou, Guinea. Sequence and phylogenetic analyses of the genetic material demonstrate a novel hantavirus species, which we propose to name "Sangassou virus." PMID- 16704851 TI - Heterogeneity among Mycobacterium ulcerans isolates from Africa. AB - Mycobacterium ulcerans causes Buruli ulcer, an ulcerative skin disease in tropical and subtropical areas. Despite restricted genetic diversity, mycobacterial interspersed repetitive unit-variable-number tandem repeat analysis on M. ulcerans revealed 3 genotypes from different African countries. It is the first time this typing method succeeded directly on patient samples. PMID- 16704852 TI - Human Bocavirus infection, Canada. AB - Human Bocavirus was detected in 18 (1.5%) of 1,209 respiratory specimens collected in 2003 and 2004 in Canada. The main symptoms of affected patients were cough (78%), fever (67%), and sore throat (44%). Nine patients were hospitalized; of these, 8 (89%) were <5 years of age. PMID- 16704853 TI - Lymphocytic choriomeningitis in Michigan. AB - We summarize the first reported case of acquired lymphocytic choriomeningitis virus (LCMV) infection in Michigan to be investigated by public health authorities and provide evidence of the focal nature of LCMV infection in domestic rodents. Results of serologic and virologic testing in rodents contrasted, and negative serologic test results should be confirmed by tissue testing. PMID- 16704854 TI - Second human case of Cache Valley virus disease. AB - We document the second known case of Cache Valley virus disease in a human. Cache Valley virus disease is rarely diagnosed in North America, in part because laboratories rarely test for it. Its true incidence, effect on public health, and full clinical spectrum remain to be determined. PMID- 16704855 TI - [Spontaneous growth of girls with Turner's syndrome until 6 years of age]. AB - BACKGROUND: Turner's syndrome (TS) is one of the most common genetically conditioned diseases, its incidence amounting to one per 1900-2500 live female births. A marked short stature is a prime symptom of the disease, the average adult body height of untreated women being by 23 cm lower than that of healthy ones. According to Ranke, developmental disorders appear as early as in foetal life and are responsible for body length and mass deficits at birth in relation to gestational age. OBJECTIVES: To revise present views regarding the course of physical development of girls with TS, especially at infancy and early childhood. MATERIAL AND METHODS: A total of 474 female newborns with Turner's syndrome were studied, the pregnancies, mostly second ones, not less than 38 weeks. Turner's syndrome was confirmed by chromosome analysis. Body mass at birth (BM) was related to the norms for gestational age (GA) designed by Usher and McLean. RESULTS: Growth disorders take place in TS since infancy, the growth rates in the age period of 2-3 years being similar to those observed in healthy population. After that period, however, growth rate in Turner girls steadily decreases as shown in percentile norms for TS. At the age of 6 years, the average difference in body height between Turner and healthy girls exceeds 1/3 of the final growth deficit noted in untreated adult women with TS. CONCLUSION: We have demonstrated that body mass and length of Turner newborns were normal in relation to gestational age. Only in about 5% of newborns a simultaneous length and weight deficit exceeded -2 SD, and that deficit was attributed to intrauterine growth retardation (IUGR). In clinical practice, newborns without documented decrease in foetal growth rate, but having a pronounced body length deficit, are diagnosed as IUGR-afflicted. Since simultaneous body length and mass deficits were rare in our newborns, IUGR was regarded as unlikely in bringing about growth deficit in Turner girls. In about 1/5 of Turner newborns a weight or length deficit was found (19 and 1%, respectively). Since either of those deficits evidences intrauterine dystrophy (small for gestational age, SGA), the latter one should be considered much more frequent in TS than in the general population. Therefore, SGA appears to be one of the symptoms of TS and should thus be taken into account when explaining body mass and/or length deficit in female newborns. Growth disorders in foetuses with chromosomal aberrations typical of TS are, in all probability, as frequent as in those with normal both X-chromosomes. PMID- 16704856 TI - [Pituitary response to GnRH analogue testing in girls with a polycystic ovary syndrome]. AB - THE AIM of our study was to estimate the gonadotropin level after GnRH analogue injection in girls with PCOS after suppression with dexamethasone. MATERIAL AND METHODS: 57 girls with hirsutism, mean age 15.9 years, were involved in the study. The research was performed in the early and middle follicular stage. Menstrual disorders were observed in 78% of them. The patients were divided into 3 groups: I -- with clinical and laboratory symptoms of PCOS (menstruation disorders, testosterone >65 ng/ml and/or LH/FSH >2; n=29), II -- with menstruation disorders and without elevated androgen level (n=15), III -- without menstruation disorders and without elevated androgen level (n=13). Basal blood samples were drawn at 8 a.m. GnRH analogue (Relefact LH-RH) 100 microg was then given subcutaneously and blood samples were drawn every 4 hours for 24 hours. RESULTS: Basal level of LH was the highest in group I (6,18+/-4,10 IU/l) in comparison with II (5.53+/-3.40 IU/l) and III (3.82+/-2.79 IU/l). After GnRH analogue administration mean LH concentration increased in all groups and peaked after 2 hours. Stimulated LH level was the highest in group I and differed statistically significantly from group III during the whole period of the test. The most significant difference occurred at 12 a.m. (p=0.003) and 10 a.m. (p=0.004). The FSH secretion in all tested groups was similar. It peaked, like LH, after 2 hours after GnRH analogue injection and decreased slightly during next 2 hours. A marked decrease was observed in the following period of time. CONCLUSIONS: 1. High and fast LH secretion responding to GnRH analogue indicates masculinization of the hypothalamo-pituitary axis in PCOS girls. 2. The hirsute girls without menstrual disturbances and hormonal abnormalities probably also have subtle masculinization of the pituitary response to stimulation by GnRH analogue. PMID- 16704857 TI - [Influence of fat tissue distribution on metabolic complications in children and adolescents with simple obesity]. AB - BACKGROUND: Increased prevalence of obesity in children and adolescents results in more common metabolic complications characteristic for adults, particularly those with abdominal obesity. The objective of the study was to determine the relation between the fat tissue distribution and metabolic complications and to estimate the prevalence of the metabolic syndrome in obese children and adolescents. MATERIAL AND METHODS: We studied 64 children (42 girls and 22 boys) with simple obesity (BMI =97 pc) in the mean age 12.4+/-3.4 years. The fat tissue distribution was assessed on the basis of waist circumference, hip circumference, waist to hip ratio (WHR) and skinfold thickness (suprailiac, subscapular, biceps and triceps). In all children plasma concentrations of total cholesterol, HDL and LDL cholesterol as well as triglycerides were estimated. Plasma glucose and insulin levels were measured in fasting state and during the oral glucose tolerance test (OGTT). Fasting insulin to glucose ratio (FIGR) was calculated. Blood pressure was measured in triplicate. RESULTS: In 33 (51.6%) of children dyslipidemia, in 10 (15.6%) hyperinsulinemia or impaired glucose tolerance and in 12 (18%) hypertension was diagnosed. The Metabolic syndrome was present in 9 (14%) children. The anthropometric predictor for the risk of metabolic complications was a greater waist circumference, while greater hip circumference decreased the risk. CONCLUSIONS: The metabolic complications characteristic of metabolic syndrome, previously diagnosed exclusively in adults, may occur also in obese children and adolescents. As in adults, abdominal obesity is the most relevant risk factor of the metabolic syndrome. PMID- 16704858 TI - [Evaluation of selected risk factors of atherosclerosis in children with type 1 diabetes mellitus and hypercholesterolemia]. AB - BACKGROUND: Increased risk of atherosclerosis in individuals with diabetes mellitus is not fully explained by the conventional risk factors. Therefore, new independent risk factors are taken under consideration. AIM OF THE STUDY was to evaluate selected risk factors of atherosclerosis in children with type 1 diabetes mellitus and hypercholesterolemia. MATERIAL AND METHODS: The study comprised 30 diabetic children, aged 5-18 years with plasma cholesterol concentration exceeding 170 mg/dl. In children, their parents and siblings plasma concentration of total cholesterol, HDL and LDL-cholesterol, triglicerides, homocysteine, apolipoprotein A-I, apolipoprotein B was measured and electrophoretic distribution of lipids was performed. Anthropometric measurements with BMI estimation were performed in diabetic children. Data were analyzed with respect to BMI, metabolic control and physical activity in children and with respect to family history of atherosclerosis. RESULTS: No significant differences in measured parameters were found between diabetic children and their healthy siblings besides significantly lower homocysteine concentration (p=0.037). Concentration of triglicerides was significantly higher in children with low than with high physical activity (101.6+/-64.6 mg/dl vs. 60.2+/-27.4 mg/dl respectively, p<0.05), HDL cholesterol was significantly lower in children with poor than with a satisfactory metabolic control (58.3+/-10.0 mg/dl vs. 69.1+/-12 mg/dl, p<0.05) and lipoprotein apoB concentration was significantly higher in children with positive (0.77+/-0.15 g/l) compared to children with a negative family history of atherosclerosis (0.66+/-0.09 g/l, p<0.05). Overweight did not influence the measured parameters significantly. Statistically significant positive correlations were found for apoB between affected children and their mothers (r=0.62), for LDL-cholesterol between affected children and their fathers (r=0.35), and for total cholesterol (r=0.47) and homocysteine (r=0.44) between affected children and their siblings. CONCLUSIONS: 1. Lipid disorders in children with type 1 diabetes mellitus may be present regardless their metabolic control, but they are more pronounced in children with poor control 2. Higher triglicerides concentrations were found in children with low physical activity and higher apolipoprotein B concentration in children with a positive family history for atherosclerosis. 3. Parental lipoprotein concentration may significantly influence the lipid profile in their diabetic offspring. PMID- 16704859 TI - [Evaluation of final height in patients with pituitary growth hormone deficiency who were treated with growth hormone replacement]. AB - BACKGROUND: Growth hormone (GH) therapy has been used for children with pituitary GH deficiency. It resulted in improving their height velocity and achieving an adult height in the normal range for the general population. OBJECTIVES: To evaluate the final height in childhood-onset growth hormone deficiency patients who had already completed treatment and were still GH deficient in adult life. MATERIAL AND METHODS: 21 children (12 boys and 9 girls) diagnosed as GH deficiency and treated with growth hormone to final height at doses of 0.17 mg/kg/week (0.5 IU/kg/week) subcutaneously for 7 days. There were 7 patients with isolated GHD and 14 with multiple pituitary hormone deficiencies. RESULTS: At the diagnosis peak serum GH concentrations were 2.8+/-2.8 mU/l in insulin tolerance test and 3.3+/-2.2 mU/l in clonidine test. Reconfirmation of the GH deficiency diagnosis after growth hormone treatment revealed a peak serum GH 1.77+/-1.2 mU/l in insulin tolerance test. Mean chronological age of the patients at the beginning of treatment was 10.29+/-3.57 years and was significantly higher in boys. Patients had completed a course of treatment in the chronological age of 17.85+/-1.97 years. Children began treatment with mean bone age 7.24+/-3.57 years and ended with 15+/-0.97 years. After the treatment a significant improvement in height was shown. Height SDS at the beginning of the treatment was -4.03+/-0.91 and -0.69+/-1.01 after the treatment. There was no difference between final height and target height (-0.54+/-0.93 SDS) in our patients. CONCLUSIONS: Children with pituitary growth hormone deficiency who were treated with growth hormone replacement achieve a final height in the normal range for the general population and their target height. PMID- 16704860 TI - [Current views on the etiopathogenesis of goiter in children]. AB - The most frequent cause of goiter in children is a deficit of iodine, leading to endemia of goiter in the regions with insufficient supplementation of this element. Goiter occurs also in the course of autoimmunological diseases of the thyroid gland (Hashimoto disease, Graves' disease), genetically-related disorders of thyroid hormones, biosynthesis/impaired biosynthesis of thyroid hormones. According to the theory of goiter pathogenesis, excessive enlargement of the thyroid gland is due to adaptation of follicle cells of the gland aiming at neutralizing the impaired synthesis of the thyroid hormones caused by various intrathyroid, environmental and genetic factors/agents. The mechanisms stimulating thyrocytes to hyperplasia or hypertrophy are very complex and still unknown in spite of having identified many physiological and pathogenetic factors connected with goiter. PMID- 16704861 TI - [Guidelines concerning insulin dosage in children and adolescents with type 1 diabetes on continuous subcutaneous insulin infusion]. AB - The treatment of diabetes mellitus with a continuous subcutaneous insulin infusion (CSII) has become very popular and is well accepted by diabetic patients. Pump therapy mimics the physiological insulin secretion and has been shown to be a safe and effective method of insulin administration alternative to the method of multiple injections. Continuous insulin infusion provides greater flexibility in the timing of meals and snacks with higher treatment satisfaction. Programmed basal rates improve nocturnal glycemic control and help to minimize a pre-breakfast increase of blood glucose level (the dawn phenomenon). Moreover, CSII can reduce exercise-induced and nocturnal hypoglycemia. Insulin pump therapy is effective in lowering glycated hemoglobin levels without higher risk of severe hypoglycaemia and ketoacidosis. To achieve a proper metabolic control with this method of treatment, the patient and his family requires appropriate education including knowledge of diet management, insulin therapy and manual competence of pump device. Here we present general guidelines for patients education concerning insulin dosage, programming of basal insulin rates and meal boluses based on carbohydrates and protein-fat exchanges. PMID- 16704862 TI - [Familial precocious puberty -- a variant of norm or pathology?]. AB - Since 1937, when familial precocious puberty (FPP) was described for the first time, only few reports on FPP have been published. The majority of them is concerned with the most investigated form of FPP, occurring only in male - testotoxicosis (male-limited precocious puberty -- MLPP). Recently another form of FPP -- familial hyperestrogenism (aromatase excess syndrome -- AES) has been described. The authors aimed to review the literature data concerning different forms of FPP emphasizing the diagnostic criteria, etiology, mode of inheritance and treatment. PMID- 16704863 TI - [Growth failure in a boy with Klinefelter syndrome and IUGR]. AB - BACKGROUND: Children born with a history of Intra-Uterine Growth Retardation (IUGR) may be susceptible for reduced growth velocity and growth deficiency. The causes of GH/IGF-1 resistance are unknown. Klinefelter syndrome is characterized by excessive growth, resulting from hypogonadism, open bone age and a prolonged growth. OBJECTIVES: The case of a 6-year-old boy with Klinefelter syndrome born with IUGR and deep growth deficiency was studied. A CASE REPORT: A 6-year-old boy with birth weight 2300 g born of term in 38 Hbd is presented. The cariotype was 47XXY. The phenotype was typical for a child with IUGR. GH in the clonidin test was 37.4 microIU/ml, IGF-1 -- 111 ng/ml. Increased IGF-1 level from 178.1 ng/ml to 360.9 ng/ml was founded in the IGF-1 generation test. CONCLUSIONS: The cause of growth deficiency in this case could be GH/IGF-1 resistance. PMID- 16704864 TI - [Precocious puberty caused by McCune-Albright syndrome in a girl aged 6 years and 9 months]. AB - The McCune-Albright syndrome is characterised by polyostotic fibrous dysplasia, "cafe-au-lait" spots and autonomous hyperfunction of various endocrine organs. The authors present the case of a girl at the age of 6 years 9 months with precocious puberty (thelarche III, menarche). High estradiol level (204 pg/ml) and low gonadoptopins concentration as well as their level after GnRH administration suggested ovarian autonomy. Ovarian cysts were found on pelvic ultrasound. Other endocrine abnormalities were excluded. Single "cafe-au-lait" spot was found on the patient skin. Scintigraphy, radiography and computed tomography scans showed fibrous dysplastic bones in the right tibia and in maxillary and sphenoid sinuses. PMID- 16704865 TI - [Fibrous dysplasia of bone in a 12-year old girl]. AB - Fibrous dysplasia of bone is a rare congenital bone disease, usually coming out before the age of 30. It is 2.5% of all bone tumours and 7.5 % of benign tumours of bones. The authors present a case of a 12-years old girl with fibrous dysplasia of bone admitted to the Department with hyperparathyroidism suspicion. Pathological changes were localized in the orbital cavity and sinuses. The diagnosis was established on the basis of the clinical course, imaging and histopathological examinations. The therapy with pamidronian acid was applied. PMID- 16704866 TI - Medium- and long-term reproducibility of self-reported exposure to physical ergonomics factors at work. AB - INTRODUCTION: The literature is sparse on reproducibility of self-reported exposure to physical ergonomics risk factors for musculoskeletal disorders (MSDs). Aims of this study were to evaluate, in a cohort of workers interviewed up to three times: 1-year test-retest reliability; and 5- and 6-year recall of physical exposures. We also examined whether reproducibility was influenced by the presence of UE MSD or by technological changes introduced between the last two surveys. METHODS: A cohort of automobile manufacturing employees was interviewed at baseline, one and six years later about work history, physical and psychosocial exposures at work, upper limb symptoms, injury and medical history, and demographics. Agreement between interviews was evaluated by intraclass correlation and Spearman coefficients. Differences in exposure between 1- and 6 year follow-up were analyzed by Wilcoxon matched-pairs signed-ranks test. RESULTS: Large and significant decreases in work pace and physical effort were observed from baseline, although an upper extremity composite index was quite stable in the total population. One-year test-retest reliability was fair to good for the composite exposure index (ICC=0.58), whole-body vibration, handling parts, and tool use, but poor for the other variables considered. Long-term reproducibility, from baseline or 1-year follow-up to 6-year follow-up, was poor for the composite index and almost all single items. UE MSD case status influenced 1-year test-retest reliability, with subjects who changed case status from baseline displaying higher reliability, but not reproducibility of recalled exposures. A strong regression to the mean effect was observed on exposures reported at follow-up surveys. CONCLUSIONS: Recalled ergonomics exposures could be employed in retrospective cohort studies as a somewhat reliable and unbiased estimate of the self-reported exposures that would have been obtained up to one year earlier, but not over a longer period (5-6 years). These longer-term results may have been limited by difficulty in matching jobs between interviews; also the regression to the mean effect likely contributed to reduce agreement. Changes in production technology and work organization produced a decrease in physical workload intensity and job pace, but did not have a substantial impact on an exposure index for the upper limb. PMID- 16704867 TI - Lipopolysaccharide induced-in vivo increases in beta-defensins of the rat parotid gland. AB - Antimicrobial beta-defensins are thought to protect epithelial surfaces. Their mobilization in response to inflammation was studied in the rat parotid gland using an ELISA assay. Bacterial lipopolysaccharide (LPS), injected into the parotid duct on one side, induced a marked local inflammatory response in the parotid gland as judged by several fold increases in myeloperoxidase activity and, in histological sections, infiltration of neutrophils. Three hours after the injection, beta-defensin 1 and 3 were increased (by 41% and 15%, respectively, P<0.01) as compared to the contralateral gland. Though still elevated 6h after the injection, the percentage figures for beta-defensin 1 were, at this time, somewhat lower (30%) compared to the situation at 3h, while those for defensin 3 were significantly higher 65% (P<0.01); neither at the early nor at the late time of observation were any changes in the level of beta-defensin 2 observed. The beta-defensins under study were not detected in submandibular and sublingual glands, neither were they detected in the inflamed submandibular gland, showing also here several fold increases in myeloperoxidase activity and, in addition, the presence of inflammatory cells, following ductal injection of LPS towards the gland. PMID- 16704868 TI - Improved properties of the non-covalent coating with N,N-didodecyl-N, N dimethylammonium bromide for the separation of basic proteins by capillary electrophoresis with acidic buffers in 25 microm capillaries. AB - Capillaries (25microm I.D.) treated with the double-alkyl-chain cationic surfactant N,N-didodecyl-N, N-dimethylammonium bromide (DDAB) in an improved coating procedure were used for separation of four basic proteins in volatile buffers (ammonium acetate and ammonium hydroxyacetate) as well as in a non volatile buffer (sodium phosphate) at pH 4. The DDAB coating was stable enough to, without recoating, permit consecutive separations of the proteins up to 9h with good precisions in peak areas (RSD=1.1%) and migration times and with high apparent efficiencies (over 1 million theoretical plates/m) in the presence of a strong anodic electroosmosis. Adsorption of the proteins onto the capillary surface, which in previous studies was found to give a certain contribution to zone broadening, was eliminated with the new modified coating method. Complex formation between the proteins and phosphate buffer was studied and confirmed, and it is proposed that slow protein-buffer component interactions are the main contributions to zone broadening in protein separations by CE. PMID- 16704869 TI - Procedure for and results of simultaneous determination of aromatic hydrocarbons and fatty acid methyl esters in diesel fuels by high performance liquid chromatography. AB - The content of aromatic hydrocarbons in diesel fuels is regulated by appropriate standards, and a further reduction in the allowed concentration of these hazardous substances in these fuels is expected. The content of aromatic hydrocarbons in diesel fuels is most often determined using standard methods EN 12916 or ASTM D-6591. The content of polycyclic aromatic hydrocarbons (PAHs) is determined from a single peak obtained using normal phase high-performance liquid chromatography (NP-HPLC), a column of the NH2 type, n-heptane as the eluent, refractive index detector (RID) and backflushing of the eluent. However, the methods mentioned above cannot be applied when the fuel contains fatty acid methyl esters (FAME), which lately has become more common. The content of FAME in diesel oils is determined using mid-IR spectrophotometry based on the absorption of carbonyl group. However, no standard procedure for the determination of classes of aromatic hydrocarbons in diesel fuels containing FAME is yet available. The present work describes such a modification of methods EN 12916/ASTM D-6591 that provides a simultaneous determination of individual groups of aromatic hydrocarbons, total content of polycyclic aromatic hydrocarbons and the FAME content in diesel fuels. The refractive index detector (RID) and n heptane as the mobile phase are still used, but backflushing of the eluent is applied after the elution of all polycyclic aromatic hydrocarbons. Additionally, ultraviolet diode array detection is used for the exact determination of low contents of polycyclic aromatic hydrocarbons and to confirm the presence of FAME in the analyzed fuel. PMID- 16704870 TI - Alkylphenol retention indices. AB - A novel type of retention indices for alkylphenols and related compounds are proposed. The alkylphenol retention indices (APRI) use para-substituted n alkylphenols as reference series. APRI for para-n-alkylphenols are per definition equal to the number of carbon atoms in the alkyl substituent; the value for phenol is zero. Application of the APRI system with different types of derivatisation of the phenolic hydroxy group showed that the derivatisation has limited influence on these indices. Especially para-substituted alkylphenols gave APRI values that could be transferred with high accuracy from one type of derivative to another. By comparing results obtained with different gradients in temperature-programmed GC, it was also shown that APRI is less affected by chromatographic conditions than retention indices based on n-alkanes. PMID- 16704871 TI - A simple, convenient and chemoselective formylation of sterols by Vilsmeier reagent. AB - Vilsmeier reagent (DMF-POCl(3)) was used as an efficient formylating agent. Several sterols having sec-hydroxyl group at 3/17-position have been modified into respective formate esters. The method is simple, mild, chemoselective and provides sec-alcoholic protection in good yields. PMID- 16704872 TI - Estrogen conjugation and antibody binding interactions in surface plasmon resonance biosensing. AB - Thioether-linked 3-mercaptopropionic acid derivatives of 17beta-estradiol and estrone were formed at the A-ring 4-position of the steroids by substitution of their 4-bromo analogues. The carboxylic acid terminal was used to link to an oligoethylene glycol (OEG) chain of 15-atoms in length. The OEG derivative of 17beta-estradiol was then in situ immobilized on a carboxymethylated dextran coated gold sensor surface used to detect refractive index changes upon protein binding to the surface by surface plasmon propagation in a BIAcore surface plasmon resonance (SPR) instrument. Two other estradiol-OEG derivatives with Mannich reaction linkage at the 2-position and hemisuccinate linkage at the 3 position were also immobilized on the sensor surfaces for comparison. Binding performance between these immobilized different positional conjugates and monoclonal anti-estradiol antibody, raised from a 6-position conjugate, clearly demonstrated that both 2- and 4-conjugates, not conjugated through existing functional groups, gave strong antibody bindings, whereas the 3-conjugate through an existing functional group (3-OH) gave very little binding (2% compared to the 2-conjugate). Both 2- and 4-position conjugates were then applied in a highly sensitive estradiol SPR immunoassay with secondary antibody mediated signal enhancement that gave up to a 9.5-fold signal enhancement of primary antibody binding, and a detection limit of 25 pg/mL was achieved for a rapid and convenient flow-through immunoassay of estradiol. PMID- 16704873 TI - Development of a convective diffusion model for lead pipe rigs operating in laminar flow. AB - As part of achieving lower lead standards water undertakers are utilising lead pipe rigs to quantify the benefit of treatment measures. A convective diffusion model is developed for lead pipe rigs operating in laminar flow, and applied to the three operating steps of flushing, sampling and stagnation. The model is used to determine the appropriate time-scales for each stage, and the sensitivity of the measure to variations in flow-rate. In contrast to rigs operating in turbulent flow the average lead observed leaving the pipe and that in the pipe, after a period of stagnation, are substantially different. Equations are derived for both, and take into account the residual distribution of lead left in the pipe after flushing. It is shown that the lead concentration observed leaving the pipe is well approximated by a single exponential term in contrast to the concentration within the pipe. Predictions are made on the residual lead concentration that can be achieved through flushing, and its dependence on flow rate. The relevance of the laminar flow model to that in domestic lead pipes is discussed. PMID- 16704874 TI - Interpretation of normal plasma mixing studies in the laboratory diagnosis of lupus anticoagulants. AB - INTRODUCTION: Mixing studies are part of the laboratory diagnosis of lupus anticoagulants (LA). They are used to determine the evidence of an inhibitor by demonstrating persistence of an abnormal clotting time by mixing patient plasma with normal plasma. Since there is no standardised interpretation of results of mixing studies, percent correction formulas are proposed. The sensitivity of mixing studies strongly depends on the interpretation of the results. MATERIALS AND METHODS: A retrospective analysis was performed on 361 samples, 75 LA positive and 286 LA-negative samples. For all the LA-positive samples and for 181/286 LA-negative samples mixing tests on one or more screening tests were performed. A percent correction formula and the Rosner index were calculated for all mixing tests on aPTT and dPT. RESULTS: The <70% correction formula for the mixing tests on aPTT showed the highest sensitivity (95%). The Rosner index had a sensitivity (93%) comparable with the <70% correction formula. dPT is shown to be less sensitive in the detection for LA and, even when the screening test is prolonged, interpretation of the mixing test by the percent correction formula misclassifies many samples. Rosner index in the interpretation of mixing tests for dPT is more sensitive than the percent correction formula, 49% and 57%, respectively. CONCLUSIONS: This study demonstrates that the Rosner index and the percent correction formulas for interpretation of mixing studies are complementary and can help to reduce misclassification of LA-positive or LA negative samples. PMID- 16704875 TI - Bovine model of reproductive aging: response to ovarian synchronization and superstimulation. AB - The responsiveness of the hypothalamo-pituitary axis to steroid treatments for ovarian synchronization and the ovarian superstimulatory response to exogenous FSH was compared in 13-14 year old cows and their 1-4 year old young daughters. We tested the hypotheses that aging in cattle is associated with: (1) decreased follicular wave synchrony after estradiol and progesterone treatment; (2) delayed LH surge and ovulation in response to exogenous preovulatory estradiol treatment; (3) reduced superstimulatory response to exogenous FSH. Higher plasma FSH concentrations (P<0.01), and a tendency (P=0.07) for fewer 4-5 mm follicles at wave emergence were observed in old cows (n=10) than in young cows (n=9). The suppressive effect of estradiol/progesterone treatment on FSH was similar between old and young cows. Although the preovulatory LH surge in response to estradiol treatment was delayed in old than young cows (P=0.01), detected ovulation times were not different. No difference in ovarian superstimulatory response was detected between age groups, but old cows (n=8) tended (P=0.10) to have fewer large follicles (>or=9 mm) 12 h after last FSH treatment than in young cows (n=7). We concluded that pituitary and ovarian responsiveness to estradiol/progesterone synchronization treatment was similar between old and young cows, but aging was associated with a delayed preovulatory LH surge subsequent to estradiol treatment. Old cows tended to have fewer large follicles after superstimulatory treatment than young cows. PMID- 16704876 TI - Synergistic effects of troglitazone in combination with cytotoxic agents in acute myelogenous leukaemia cells. AB - We have previously shown that troglitazone (TG) induces apoptosis in acute myelogenous leukaemia (AML) cell lines. Here we show that normal bone marrow and mobilized peripheral blood progenitors are highly resistant to TG at concentrations up to 100 microM, while primary cultures of AML bone marrow show significant decline in viability at >7.5 microM TG. The combination of standard cytotoxic agents (daunorubicin, etoposide, and cytarabine) with TG is synergistic in six AML cell lines, with the strongest synergy being exhibited when cells are pretreated with TG for 24h prior to addition of the cytotoxic agent. Significant declines in IC(50) for the cytotoxic agents are seen at nanomolar concentrations of TG. The in vitro synergy between TG and the cytotoxic drugs used in AML therapy provides a basis for in vivo evaluation of these combinations. PMID- 16704877 TI - Children at risk of neglect: challenges faced by child protection practitioners in Guatemala City. AB - OBJECTIVES: The objectives were to (1) delineate the definition, common forms, and perceived risk factors contributing to child neglect in Guatemala from the perspective of different stakeholders and (2) identify the challenges faced by child protection practitioners in identifying children at risk of neglect within the context of Guatemala. METHODS: A range of qualitative methods were used: semi structured interviews (SSI) with key stakeholders involved in child neglect (n=9); semi-structured interviews with professionals working in the child protection services (n=14); focus group discussions (FGDs) with professionals, voluntary workers and children (n=60), and the group consensus method with key informants (KI, n=10). Data analysis was conducted using the framework approach. RESULTS: Participants described child neglect as a complex social phenomenon combining parental omission of care and an indifferent or negative attitude towards the child compounded by governmental neglect. The main factors which were perceived as contributing to neglect were poverty, societal and cultural norms, and importantly, gender roles and relations. These contextual factors complicate the identification of neglect and raise a number of difficult dilemmas for child protection workers that are exacerbated by limited Guatemalan legislation on neglect and restricted alternative care options. CONCLUSION: Stakeholder perspectives were found to be useful for providing contextual information and highlighting associated challenges related to assessing child neglect in Guatemala. PMID- 16704878 TI - A method for a more complete in vitro Parkinson's model: slice culture bioassay for modeling maintenance and repair of the nigrostriatal circuit. AB - Slice culture model systems provide a unique opportunity to monitor and lesion brain circuits in a dish. Using a novel approach, we have generated parasagittal slices from mouse brains that preserve, throughout the culture process, the nigrostriatal circuit. These slices can be cultured for approximately 4 weeks with maintenance of normal neuronal cytoarchitecture. Application of the dopamine specific toxin 6-hydroxy dopamine (6-OHDA) induces a significant decline in tyrosine hydroxylase positive cell bodies and fibers. Using a transgenic mouse with green fluorescent protein (GFP) under the control of the tyrosine hydroxylase promoter, we have been able to visualize in real time the loss of GFP expression in the striatum of slices as a result of 6-OHDA exposure. Using these cultures we have demonstrated the feasibility of modeling cellular replacement strategies. GFP-positive embryonic stem cell-derived neuronal precursors can be tracked in real time throughout the experiment and are amenable to patch clamp recording within the slice environment. In addition, cell differentiation can be observed within these slices and the effects of morphogenetic proteins, like the extracellular matrix molecule laminin, drugs or small molecules can be observed. This unique culture system presents a new approach for modeling Parkinson's disease in vitro, and provides a potentially useful new method for screening cell and molecular therapies for neurodegenerative diseases. PMID- 16704879 TI - Accuracy of the three-window sampling method in morphometric analysis of human sural nerve. AB - Morphometry has an important role in the assessment of sural nerve biopsies as a part of early detection of structural abnormalities in peripheral nerve. Various sampling methods have been used to reduce time and effort needed in the analysis of total nerve fibers but their accuracy remains controversial. We examined the accuracy of three-window sampling method in the morphometric evaluation of human sural nerve biopsies by comparing with the total fiber quantification. Three windows (0.012 mm(2) each) were placed in every possible fascicle in the sections and data from all windows were pooled and analyzed for the number of myelinated axons, myelinated fiber diameter, axon diameter, myelin thickness, g ratio as well as myelinated fiber density. Means and ranges of the data from the two techniques were similar and the agreement was further confirmed by intraclass correlation analysis. These findings indicate that the three-window sampling method can be used to evaluate human sural nerve with accuracy. PMID- 16704881 TI - Request from the International Advisory Committee to DDBJ/EMBL/GenBank. PMID- 16704880 TI - Tissue engineering and cartilage regeneration for auricular reconstruction. AB - OBJECTIVE: This paper will provide (a) a review on current status of auricular reconstruction (b) particularly focusing on the current data about pinna reconstruction using stem cells in combination with tissue engineering. METHODS: The paper is divided into two sections. The first section presents a brief overview of the current status of auricular reconstruction. In the second section, the authors review the aspects and the current status of stem cells and tissue engineering researches related to cartilage regeneration. CONCLUSIONS: Total auricular reconstruction represents one of the greatest challenges for the ENT and Facial Plastic surgeon. The matter of auricular cartilage reconstruction is complex, and progresses in material designs as well as in stem cells field are essential. Even if this bio-technology field is promising, the progresses still are not adequate as patient expectations remain high. PMID- 16704882 TI - The effect of quality information on consumer health plan switching: evidence from the Buyers Health Care Action Group. AB - We examine the factors that lead employees to search for health plan quality information and the effect of such information on the decision to switch plans. Extending Hirshleifer and Riley's model [Hirshleifer, J., Riley, J.G., 1979. The analytics of uncertainty and information--an expositional survey. Journal of Economic Literature 17 (December (4)), 1375-1421] of the economics of information, we develop a two-equation model of quality information awareness and switching behavior. We estimate the model using data from a random sample of 651 single employees from 16 firms that are members of the Buyers Health Care Action Group, a health care purchasing coalition in the Minneapolis-St. Paul region. Our empirical results do not support either a link between quality information and switching behavior, or between perceived health plan satisfaction and switching. We do, however, find that switching is influenced by changes in premiums and whether an individual has an existing relationship with a health care provider. PMID- 16704883 TI - Rapid pyrazinamide susceptibility testing of Mycobacterium tuberculosis by flow cytometry. AB - The resurgence of tuberculosis along with the increased resistance of Mycobacterium tuberculosis has emphasized the need for timely susceptibility testing for control of the disease. Previous studies have shown that rapid susceptibility testing can be accomplished for isoniazid, ethambutol, and rifampin using the flow cytometric assays. In this study we compared the flow cytometric susceptibility assay with the BACTEC TB 460 and BACTEC MGIT 960 for pyrazinamide (PZA). There was 93% agreement between the BACTEC MGIT 960 and the flow cytometric methods for 100 microg/mL of PZA. Additionally, there was a 95% and 86% agreement between the BACTEC TB 460 and flow cytometric methods for 50 microg/mL and 100 microg/mL of PZA, respectively. These findings show that susceptibility testing by the flow cytometric assay is accurate. Most importantly, susceptibility results by the flow cytometric assay were available 24 h after initiation of the testing procedure. The advantages of simplicity, speed and accuracy make the flow cytometric susceptibility assay an immediate impact technology to improve patient care. PMID- 16704884 TI - Inhibition of PKR by RNA and DNA viruses. AB - Interferons were the first of the anti-viral innate immune modulators to be characterized, initially characterized solely as anti-viral proteins [reviewed in Le Page, C., Genin, P., Baines, M.G., Hiscott, J., 2000. Inteferon activation and innate immunity. Rev. Immunogenet. 2, 374-386]. As we have progressed in our understanding of the interferons they have taken a more central role in our understanding of innate immunity and its interplay with the adaptive immune response. One of the key players in function of interferon is the interferon inducible enzyme, protein kinase (PKR, activatable by RNA). The key role played by PKR in the innate response to virus infection is emphasized by the large number of viruses, DNA viruses as well as RNA viruses, whose hosts range from insects to humans, that code for PKR inhibitors. In this review we will first describe activation of PKR and then describe the myriad of ways that viruses inhibit function of PKR. PMID- 16704885 TI - Predictors of glycemic control on insulin pump therapy in children and adolescents with type I diabetes. AB - OBJECTIVE: To determine variables predictive of glycemic control in a large population of pediatric patients with type 1 diabetes on continuous subcutaneous insulin infusion (CSII). METHODS: Charts of patients on CSII for > or =1 year were reviewed. "Good" control was a priori defined as HbA1c < or =9% in patients under 12 years of age, and < or =8% in patients over 12 years. RESULTS: Ninety three patients were identified (57 girls and 36 boys). Their mean age at pump start was 11.6+/-3.1 years with duration of diabetes of 4.7+/-3.1 years. Average time on pump therapy was 2.4+/-0.8 years. HbA1C decreased from 8.7+/-0.9% prior to pump therapy to 8.3+/-0.6% while on CSII (p < 0.01). Despite analysis of a large number of possible predictors, only number of basal rates (4.4 versus 3.4) and younger age (10.0 years versus 13.1 years) correlated with good control. CONCLUSION: Only younger age and use of more basal rates were predictive of good diabetes control in children using CSII. Decisions regarding which pediatric patients are most appropriate for CSII must continue to be individualized. PMID- 16704886 TI - Equity in resource allocation for health: a comparative study of the Ashanti and Northern Regions of Ghana. AB - Debate over the equitable allocation of financial resources in the health sector has focused overwhelmingly on allocation from national to regional levels. More equitable allocation of such resources within regions has been virtually ignored, creating a vacuum in knowledge regarding how resources are allocated intra regionally and their potential influence on promoting health equity. In this paper, we report an empirical study examining progress towards equity in intra regional resource allocation in the Ashanti and Northern regions of Ghana. Relative deprivation across the 31 districts of the two regions was measured as a proxy of health needs. The result was used to develop an equity-adjusted share index (EAS) applied as a yardstick against which progress towards equity was assessed. The study found a significant correlation between districts' share of donor pooled funds (DPF) and the EAS in the Northern region for three of the 4 years investigated. In Ashanti region, a worsening trend in relation to equity in DPF allocation was discovered. The proportion of variance in the share of DPF that could be explained by the EAS reduced incrementally from 56% in 1999 to less than 1% in 2002. The study highlights the need for more emphasis on intra regional equity in resource allocation in Ghana. PMID- 16704887 TI - Correlation of the structure and conformational changes of selected fragments of plant small ribosomal RNA within the steps of polypeptide chain elongation. AB - The interaction and conformational relationships between rRNAs and ribosomal proteins are responsible for ribosome activity. We tested seven different deoxyoligonucleotides complementary to the selected, highly conserved sequences of 18S rRNAs important in protein biosynthesis. We carried out a reaction of binding Phe-tRNA to A site on the ribosomes converted either to pre- or to post translocational states (with or without pre-hybridized oligonucleotides). We found a correlation between the level of oligomer hybridization and the inhibition of AA-tRNA binding. We observed well-defined structural changes of ribosome's conformation during different steps of the elongation cycle of protein biosynthesis. PMID- 16704888 TI - The adjuvant effects of the toll-like receptor 3 ligand polyinosinic-cytidylic acid poly (I:C) on antigen-specific CD8+ T cell responses are partially dependent on NK cells with the induction of a beneficial cytokine milieu. AB - Poly (I:C), a TLR3 ligand, has shown promise as a vaccine adjuvant to CD8(+) T cell responses. The underlying mechanisms involved in creating this adjuvant response in vivo, however, have not been well defined. In this study, we explored the contribution of NK cells and inflammatory cytokines in mediation the poly (I:C) adjuvant effects. Enhanced antigen-specific CD8(+) T cell responses were observed only when poly (I:C) was administered within 4h of peptide vaccination. Poly (I:C) treatment was associated with a rapid induction of inflammatory cytokines in the serum, including IL-6, IL-10, MCP-1, TNF-alpha, IFN-alpha, and IFN-gamma, and selective increases in the numbers of NK (NK1.1(+)CD11b(+)) cells and Mvarphi (NK1.1(-)CD11b(+)), but not NK T (CD3(+)NK1.1(+)) cells. NK cells were required for the adjuvant effects of poly (I:C). Poly (I:C) treatment in TNF alpha, type I IFNR, IFN-gamma, IL-6, IL-12Rbeta2, or IL-15 gene-deficient mice revealed a reciprocal interaction and interdependence in the induction of these cytokines, where the absence of one cytokine impacted on the production of others. Further, the adjuvant effects of poly (I:C) were dependent on the endogenous levels of type I IFNs, TNF-alpha, IFN-gamma, IL-12, and IL-15. IFN alpha and IFN-beta, but not TNF-alpha or IL-6, were able to mimic the adjuvant effects of poly (I:C). We conclude that the adjuvant effects of poly (I:C) on antigen-specific CD8(+) T cells appeared to be exquisitely dependent on the rapid induction of certain beneficial cytokines produced in part by NK cells. PMID- 16704889 TI - 'Help, educate, encourage?': Geographical variations in the provision and utilisation of diabetes education in New Zealand. AB - In New Zealand, as elsewhere, it is argued that a diabetes epidemic is underway. With careful management from individuals and professionals and appropriate levels of education, it is possible to prevent many complications of diabetes. The overall objective of the paper is to evaluate the role and impact of Diabetes New Zealand (DNZ), the key voluntary sector provider of diabetes education and support services, with respect to four criteria: (i) the extent to which DNZ is reaching groups most at risk of diabetes; (ii) the degree to which it has encouraged levels of member involvement; (iii) whether voluntary group provision of education is that most preferred by members; and (iv) the extent to which members see the voluntary sector model as being effective in combating the growth of diabetes. A survey of members of six of the 41 affiliated societies of DNZ suggests that such organisations, although having a high proportion of older members, have generally failed to target more deprived groups. While the societies generally score more positively in encouraging member involvement and being perceived as effective by their members, they do not always utilise the preferred form of educational provision. However, there are significant contextual variations by urban-rural location and according to the organisational structure of the societies. Rural societies and those with decentralised organisational structures generally score highest on the above criteria. The results pose a problem for DNZ which, like many other voluntary sector organisations, is facing pressures of increased corporatisation and centralisation. We see this as an important challenge that DNZ needs to address if New Zealand is going to better cope with the emerging diabetes epidemic. PMID- 16704890 TI - What mediates the inverse association between education and occupational disability from back pain?--A prospective cohort study from the Nord-Trondelag health study in Norway. AB - Low education is consistently associated with an increased risk of back pain disability, but the underlying mechanisms for this relationship are poorly understood. In a seven-year prospective observational study of 38,426 employed men and women between 25 and 59 years in Norway, we investigated to what extent occupational class, working conditions and individual lifestyle mediated the effect of formal education on disability pensioning from back pain. Each additional year of formal education was associated with decreased risk for disability pensioning from back pain for both men [age adjusted Hazard Ratio (HR) 0.77; (95% Confidence Interval, 0.72-0.82)] and women [HR 0.76(0.71-0.82)]. Adjustment for occupational class and factors related to working conditions (authority to plan own work, physically demanding work, concentration and attention and job satisfaction) and individual lifestyle (smoking, body mass index, physical exercise and alcohol consumption) reduced the effect of education by 39% [HR 0.86(0.79-0.93)] for men and by 21% [HR 0.81(0.73-0.89)] for women. Working conditions contributed most to the explanation for men, while occupational class, working conditions and life style factors contributed equally for women. Subgroup analyses indicate small differences between full-time and part-time employees, while some differences were found between subcategories of back diseases. The study indicates that there is a strong and unexplained effect of education on back pain disability pensioning, which is not mediated by occupational class, working conditions or individual lifestyle. PMID- 16704891 TI - Fulminant hyperammonaemia induced by thiopental coma in rats. AB - Fulminant hyperammonaemia as a threshold effect of coma-inducing dose of sodium thiopental has been revealed in rats. Blood ammonia content increased progressively after the introduction of 1.0 LD(50) (but not 0.8 LD(50)) of sodium thiopental three times in 3h and five times in 18h. The urinary ammonia excretion was not impaired while the volatilization of ammoniac from the body of ST-treated rats was higher, giving evidence of the augmentation of ammonia production. Blood urea increased by one third despite of insignificant alterations of haematocrit and blood creatinine. Ammonia hyperproduction in the digestive tract could result from gastrointestinal stasis, which has been verified by roentgenography and confirmed by correlation of hyperammonaemia with the stool retardation. In thiopental coma rats the slope of a dose-dependent increase of the blood ammonia and the blood urea after the intraperitoneal injection of ammonium acetate did not exceed that in intact animals. So the ammonia hyperproduction in the digestive tract could be the main contributing cause of fulminant hyperammonaemia in rats with thiopental coma and thus be involved into pathogenesis of the coma. PMID- 16704892 TI - Open vs. laparoscopic radical prostatectomy... and laparoscopy is better! PMID- 16704893 TI - Extent of prostate-specific antigen contamination in the Spanish section of the European Randomized Study of Screening for Prostate Cancer (ERSPC). AB - OBJECTIVES: The performance of tests outside prostate cancer screening trials (PSA contamination) may affect their statistical power. The present study addressed the extent of PSA contamination in the Spanish section of the European Randomized Study of Screening for Prostate Cancer (ERSPC) and its impact on biopsy performance and prostate cancer detection. METHODS: Data linkage was performed to address screening-related interventions outside the study. Four databases were used: (1) Spanish ERSPC database (n=4278), (2) laboratory database with all PSA determinations (n=31,140), (3) database of 1608 prostate biopsies, and (4) records of all prostate cancers (n=819) diagnosed at our centre. PSA contamination, biopsy performance, and cancer detection rates were calculated. RESULTS: Median follow-up time was 6.6 yr. A total of 2201 PSA determinations were performed for 1253 men. Cumulative PSA contamination was 29.3% (17% in the control arm during the first 4 yr). A higher proportion of men undergoing biopsies was found in the screening arm (21.3% vs. 2.9% in the control arm, p<0.0001). Similarly, higher cancer detection rates were found in the screening (4.7% vs. 1.2% in the control arm, p<0.0001). CONCLUSIONS: In our experience, the PSA contamination rate has increased during the last years, but its impact on biopsy performance and cancer detection in the control arm of the trial is limited and not likely to compromise the statistical power of the ERSPC trial. PMID- 16704894 TI - A randomised trial comparing holmium laser enucleation versus transurethral resection in the treatment of prostates larger than 40 grams: results at 2 years. AB - OBJECTIVE: To compare holmium laser enucleation of the prostate (HoLEP) with transurethral resection of the prostate (TURP) for treatment of men with bladder outflow obstruction (BOO) secondary to benign prostatic hyperplasia with a minimum of 24-month follow-up. PATIENTS AND METHODS: Sixty-one patients were randomised to either HoLEP or TURP. All patients had BOO proven on urodynamic studies pre-operatively (prostate size 40-200 g). One patient died before treatment, which left 30 patients in each group. Perioperative data, as well as symptom scores, Quality of Life (QoL) scores, and maximum urinary flow rates (Qmax) were obtained at one, three, six,12, and 24 months. Post-void residual volumes, transrectal ultrasound (TRUS) volumes, and pressure flow studies were obtained six months post-operatively. Continence and potency data were also recorded. RESULTS: There were no significant differences between the two surgical groups pre-operatively. Mean pre-operative TRUS volume was 77.8+/-5.6 g (42-152) in the HoLEP group and 70.0+/-5.0 g (46-156) in the TURP group. Patients in the HoLEP group had shorter catheter times and hospital stays. More prostate tissue was retrieved in the HoLEP group. At six months, HoLEP was urodynamically superior to TURP in relieving BOO. At 24 months, there was no significant difference between the two surgical groups with respect to American Urology Association scores, QoL scores, or Qmax values; however, two patients in the TURP group required re-operation. CONCLUSIONS: HoLEP has less perioperative morbidity and produces superior urodynamic outcomes than TURP, when treating prostates >40 g. At 24 months of follow-up, HoLEP is equivalent to TURP. PMID- 16704896 TI - Production of materials with alumina and ashes from incineration of chromium tanned leather shavings: environmental and technical aspects. AB - The leather tannery industry produces a significant amount of solid and hazardous wastes. Chromium-containing wastes like tanned shavings used to be incinerated in order to recover energy. The incineration process generates ashes that must be disposed of. This paper is a report on the results of the evaluation of technological properties and environmental compatibility of products made of alumina and ashes from incinerated chrome tanned shavings. The raw materials, tannery ashes and alumina were mixed together in different proportions. The ceramic bodies were molded using a hydraulic press and fired with a heating rate of 100 K/h until 1400 degrees C for 4 h in a muffle furnace. The ceramic specimens were characterized regarding physical, mechanical and thermal properties. Leaching tests, according to Brazilian, German and Dutch regulations, were performed on ceramic bodies containing different additions of ash. Results show that the ceramic materials produced are acceptable for refractory applications. PMID- 16704895 TI - Recombinant activity-dependent neuroprotective protein protects cells against oxidative stress. AB - Activity-dependent neuroprotective protein (ADNP) is essential for brain formation. Here, we investigated the potential neuroprotective effects of recombinant ADNP under stress conditions. The human ADNP cDNA was sub-cloned into a vector that contains VP22, a Herpes virus protein that may allow penetration of fused proteins through cellular membranes. When incubated with pheochromocytoma (PC12) cells, a neuronal model, VP22-ADNP was associated with the cells after a 25-min incubation period. Pre-incubation with VP22-ADNP enriched protein fractions protected against beta amyloid peptide toxicity and oxidative stress (H2O2) in PC12 cells. VP22 by itself was devoid of protective activity. Furthermore, the pro-apoptotic protein p53 increased by 3.5-fold from control levels in the presence of H2O2, while treatment with VP22-ADNP prior to H2O2 exposure significantly reduced the p53 protein levels. ADNP expression was previously shown to oscillate as a function of the estrus cycle in the mouse arcuate nucleus, these oscillations are now correlated with increased cellular protection. PMID- 16704897 TI - Particulate polycyclic aromatic hydrocarbons and n-alkanes in recycled paper processing operations. AB - The aliphatic and polycyclic aromatic fractions of dust collected in the vicinity of recycled paper processing operations were analyzed using gas chromatography/mass spectrometry. Total measured dust concentration (up to 8.73+/ 2.02 mg/m3) fluctuated substantially in the different steps of paper manufacture. This was attributed to the type of the operation such as, the release of fibers and particles during paper cutting and the use of reactive chemicals and excessive water under high temperature and pressure during the bleaching and the water rinsing/drying step. Particulate polycyclic aromatic hydrocarbons (PAHs) (from fluorene to benzo[ghi]perylene with mean concentrations from 3.8+/-0.5 to 41.4+/-0.4 ng/m3) and the unresolved mixture of branched, cyclic and unsaturated hydrocarbons (UCM) were measured in all samples while, n-alkanes from n-C20 to n C27, were only observed in cutting and packaging areas (from 180.6+/-18.9 to 4297.9+/-794.9 ng/m3). The profile of occupational PAHs in bleaching and treatment of contaminated recycled raw paper was comparable to that observed for anthropogenic activities. The values of molecular diagnostic ratios indicated the contribution of oil residues and emissions from worklifts and other equipment used for pulp and paper handling. Total benzo[a]pyrene-equivalent (BaP) concentrations of particulate PAHs (varied from 323 up to 1104 pg/m3), provided evidence that workers were exposed to high quantities of PAHs as compared with exposures to urban air and other indoor settings, posing long-term threat to their health. PMID- 16704898 TI - Solar assisted photocatalytic and photochemical degradation of Reactive Black 5. AB - The photocatalytic oxidative degradation of Reactive Black 5 (RB 5) has been investigated using TiO(2)-P25 as photocatalyst and sunlight as irradiation source in slurry form. The degradation was carried out at different experimental conditions to optimize the parameters such as amount of catalyst, concentration of dye and pH. A complete degradation of 3.85 x 10(-4) M dye solution under solar irradiation was observed in 3.5 h. The photochemical degradation using hydrogen peroxide results in the partial removal of the dye. PMID- 16704899 TI - Improved photocatalytic activity of zeolite- and silica-incorporated TiO2 film. AB - Porous TiO2 film was prepared by sol-gel method from TiO2 sol containing polyvinylpyrolidone (PVP). Photocatalytic activity of the film was evaluated by the elimination rate of ethylene. Several adsorbents including zeolite and silica powders were incorporated into the TiO2 film. All the adsorbents enhanced the activity. The optimum adsorbent content was 0.005-0.01 g/ml of the coating sol solution. Silica provided better activity than zeolite. At high humidity and in dry air the activity decreased. PMID- 16704900 TI - Pyrolysis of scrap tyres with zeolite USY. AB - A zeolite catalyst of ultrastable Y-type (USY) was investigated in the research of two staged pyrolysis-catalysis of scrap tyres. Scrap tyres were pyrolysed in a fixed bed reactor and the evolved pyrolysis gases were passed through a secondary catalytic reactor. The main objective of this paper was to investigate the effect of zeolite USY on the yield of products and the composition of derived oil. The influences of several parameters such as pyrolysis temperature, catalytic temperature, catalyst/tyre ratio, heating rate, etc. on the yield of the derived oil, char and gas were investigated. It showed that the increase of catalytic temperature and catalyst/tyre ratio resulted in high yield of gas at the expense of the oil yield. For example, when the catalyst/tyre ratio increased from 0.25 to 1.0, the yield of gas increased from 30.5 to 49.9 wt.%, and the oil yield decreased nearly two-fold from 31.6 to 12.7 wt.%. The concentration of light naphtha (boiling point < 160 degrees C) was also investigated in this study. And the high catalyst/tyre ratio favored to increase the concentration of light naphtha (< 160 degrees C) in oil. In order to study the composition of derived oil, a distilled fraction (< 280 degrees C), which was 92.5 wt.% of the oil obtained from catalytic pyrolysis of scrap tyre at a pyrolysis temperature, catalytic temperature and catalyst/tyre ratio of 500, 400 degrees C and 0.5, respectively, was analyzed with gas chromatography/mass spectrometry (GC/MS). The distillate was found to contain 1.23 wt.% benzene, 9.35 wt.% toluene, 3.68 wt.% ethylbenzene, 12.64 wt.% xylenes, 1.81 wt.% limonene and 13.89 wt.% PAHs, etc., where the single ring aromatics represented a significant potential use as chemicals. PMID- 16704901 TI - Investigation on the sonocatalytic degradation of parathion in the presence of nanometer rutile titanium dioxide (TiO2) catalyst. AB - The nanometer rutile titanium dioxide (TiO2) powder was adopted to act as the sonocatalyst after treatment of high-temperature activation and the ultrasound of low power was used as an irradiation source to induce heat-treated TiO2 powder performing sonocatalytic degradation of parathion. Although there are many factors influencing sonocatalytic degradation of parathion, the experimental results demonstrate that the optimal degradation condition of parathion can be obtained when the experimental conditions such as initial concentration of 50 mg/L parathion, addition amount of 1000 mg/L nanometer rutile TiO2, ultrasonic of 30-50 kHz frequency and 50 W output power, acidity of pH 10.0 and temperature of 20 degrees C are adopted. The degradation ratio of parathion surpassed 90% within 120 min ultrasonic irradiation in these optimal experiment conditions. The total degradation process of parathion has been monitored by UV-vis spectra and ion chromatography. At last, the parathions in aqueous solution are completely degraded and become some simple inorganic ions such as NO3(-), PO4(3-), SO4(2-), etc. In addition, the sonocatalytic activities of reused TiO2 catalysts were also studied and found to decline gradually along with the reused times. In this paper, the research on sonocatalytic degradation kinetics was also been performed and found to follow pseudo first-order reaction. All experiments indicated that the sonocatalytic method in the presence of nanometer rutile TiO2 powder was an advisable choice for the treatments of non- or low-transparent organic wastewaters in future. PMID- 16704902 TI - Determination of trace nickel in water samples by cloud point extraction preconcentration coupled with graphite furnace atomic absorption spectrometry. AB - A new method based on the cloud point extraction (CPE) preconcentration and graphite furnace atomic absorption spectrometry (GFAAS) detection was proposed for the determination of trace nickel in water samples. When the micelle solution temperature is higher than the cloud point of surfactant p octylpolyethyleneglycolphenyether (Triton X-100), the complex of Ni2+ with 1 phenyl-3-methyl-4-benzoyl-5-pyrazolone (PMBP) could enter surfactant-rich phase and be concentrated, then determined by GFAAS. The main factors affecting the cloud point extraction were investigated in detail. An enrichment factor of 27 was obtained for the preconcentration of Ni2+ with 10 mL solution. Under the optimal conditions, the detection limit of Ni2+ is 0.12 ng mL(-1) with R.S.D. of 4.3% (n = 10, c = 100 ng mL(-1)). The proposed method was applied to determination of trace nickel in water samples with satisfactory results. PMID- 16704903 TI - Characteristic overpressure-impulse-distance curves for vapour cloud explosions using the TNO Multi-Energy model. AB - A number of models have been proposed to calculate overpressure and impulse from accidental industrial explosions. When the blast is produced by ignition of a vapour cloud, the TNO Multi-Energy model is widely used. From the curves given by this model, data are fitted to obtain equations showing the relationship between overpressure, impulse and distance. These equations, referred herein as characteristic curves, can be fitted by means of power equations, which depend on explosion energy and charge strength. Characteristic curves allow the determination of overpressure and impulse at each distance. PMID- 16704904 TI - Safe immobilization of Cr(III) in heat-treated zeolite tuff compacts. AB - The possibility to remove chromium ions from a simulated electroplating wastewater by a discontinuous ion-exchange process based on phillipsite-rich Neapolitan yellow tuff (NYT) has been investigated. The immobilization of the pollutant cation in the resulting sludge through a heat-treatment has been realized, demonstrating that compacts made of Cr-loaded NYT powder, fired at temperatures of 1000 degrees C or over, are safe materials with negligible Cr3+ leaching. The set up overall process allows, in addition, a volume reduction of the waste with associated lower disposal costs or, better, the obtainment of a ceramic material, whose physical and mechanical properties are comparable to those of similar ordinary ceramics, such as bricks. In summary, the proposed strategy looks at the polluted sludge as a resource to be exploited, instead of a dangerous material to safely dispose of. PMID- 16704905 TI - Equilibrium and kinetic studies for the biosorption system of copper(II) ion from aqueous solution using Tectona grandis L.f. leaves powder. AB - The biosorption of copper(II) ions from aqueous solution by Tectona grandis L.f. was studied in a batch adsorption system as a function of pH, metal ion concentration, adsorbent concentration and adsorbent size. The biosorption capacities and rates of copper(II) ions onto T. grandis L.f. were evaluated. The Langmuir, Freundlich, Redlich-Peterson and Temkin adsorption models were applied to describe the isotherms and isotherm constants. Biosorption isothermal data could be well interpreted by the Langmuir model with maximum adsorption capacity of 15.43 mg/g of copper(II) ion on T. grandis L.f. leaves powder. The kinetic experimental data properly correlated with the second-order kinetic model. Various thermodynamic parameters such as deltaG(o), deltaH(o), and deltaS(o) were calculated indicating that this system was a spontaneous and exothermic process. PMID- 16704906 TI - Estimation of future outflows and infrastructure needed to recycle personal computer systems in California. AB - The objectives of the present study are to estimate future quantities of electronic waste (e-waste) for which appropriate infrastructure needs to be established, and to the estimate the total cost for e-waste recycling in California. Estimation of the future amounts of e-waste as a function of time is critical to effective e-waste management. To generate estimates, we use a time series materials flow analysis model (MFAM). The model estimates future e-waste quantities by modeling the stages of production, usage, and disposal. We consider four scenarios for the estimation of future e-waste generation in order to consider the effects of exportation outside the State of California and of user preferences to store or to recycle the e-waste. These efforts were further investigated through the use of sensitivity analysis. The results of the present research indicate that the outflow (recycling) amount of central processing units (CPUs) will increase and will reach approximately 8.5 million units per year in 2013, but the outflow (recycling) of cathode ray tube (CRT) monitors will decrease from 2004 in California because of the replacement of CRT monitors by liquid crystal display (LCD) monitors. In 2013, the cost for CPU recycling will be 1.7 times higher than that in 2005. But for CRT monitors, the cost for recycling in 2013 will be negligible. After the State of California enacted the ban on landfill disposal of e-waste, recycling became the most common end-of-life (EOL) option in California. Also, after 2005, the State of California will need more than 60 average-capacity materials recovery facilities (MRFs), to recycle the number of personal computer systems generated, which represents an investment in capital of over 16 million dollars. PMID- 16704907 TI - Adsorption of dyes and phenol from water on resin adsorbents: effect of adsorbate size and pore size distribution. AB - Batch adsorption runs of two commercial reactive dyes (methylene blue and reactive orange X-GN) and phenol from water onto two resin adsorbents (Amberlite XAD-4, a macroreticular adsorbent and ZCH-101, a hyper-cross-linked adsorbent) were carried out in the present study. Effect of adsorbate size and pore structure of the resin adsorbents on adsorption was discussed and Langmuir and Freundlich models were tested for the relevant adsorption isotherms. In the kinetic section concentration-time profiles were obtained to further elucidate the effect of adsorbate size and porous structure on adsorption mechanism. A pseudo-second-order model was proved to give a satisfactory description of the related kinetic results. Two-stage kinetic curve of methylene blue onto ZCH-101 was observed at a specific experimental condition and modeled schematically according to its specific pore size distribution. Furthermore, other kinetic experiments were performed at different ambient temperature to validate the proposed schematic model. PMID- 16704908 TI - Primary explosives: electrostatic discharge initiation, additive effect and its relation to thermal and explosive characteristics. AB - All explosives, under all conditions must be considered vulnerable to generation, accumulation and discharge of static charge. The low energy static hazards of the order as low as 2-3 mJ need to be guarded against in case of highly sensitive compounds namely primary explosives. The hazard is normally associated with manufacturing and filling operations due to discharge of static charge accumulated on a person supplying energy up to 20 mJ. To reduce the risk associated with static initiation hazard in the processing and handling of the explosives, the electrostatic sensitivity tests can provide an important input regarding electrostatic hazards. This paper presents electrostatic sensitivity data in terms of zero ignition probability data (E(SE0)) of some of the initiatory explosives such as nickel/cobalt hydrazinium nitrate, silver azide, lead azide and mercury salt of 5-nitro tetrazole. Similar data has also been presented for samples coated with polyvinyl pyrrolidone to study its effect on electrostatic sensitivity. The electrostatic spark sensitivity of some conventional and novel made to explain the increased spark sensitivity behavior on the basis of the possible primary explosives has been studied. The electrostatic spark sensitivity of primary explosives decreased in the order of AgN3 = NHN > PbN6 > MNT > CoHN > BNCP. A possible correlation of spark energy with approximation and assumption has been drawn with thermal, detonation and mechanical properties. The polyvinyl pyrrolidone coated samples followed the same order but interestingly with increased spark sensitivity. An attempt has been reasoning of dielectric nature of the materials or exothermic effects of decomposition products of PVP. The present work also reports the electrostatic spark sensitivity of cap compositions. PMID- 16704909 TI - Prevalence of DSM/ICD-defined nicotine dependence. AB - We conducted a qualitative review to estimate for adults (1) the lifetime and current prevalence of DSM/ICD-defined nicotine dependence and (2) the prevalence of individual DSM/ICD dependence criteria. Systematic computer searches and other methods located eleven population-based surveys of adults (>or=18 year olds) and two of young adults (18-30 year olds). In the USA and Germany, about 25% of adults had been dependent on nicotine in their lifetime, including 15% who were currently dependent. Similar or higher rates were seen in Asian men but <5% of Asian women had been dependent. About a third of ever-smokers and half of current smokers either had been or were currently dependent on nicotine and this did not consistently differ by age, country or sex. Impaired control over tobacco use was the most commonly endorsed criteria and giving up activities to use and spending lots of time with nicotine were the least commonly endorsed. Nicotine dependence is one of the most common mental disorders; however, about half of current smokers do not meet DSM/ICD dependence criterion. PMID- 16704910 TI - Origin, genetic diversity, and population structure of Chinese domestic sheep. AB - To characterize the origin, genetic diversity, and phylogeographic structure of Chinese domestic sheep, we here analyzed a 531-bp fragment of mtDNA control region of 449 Chinese autochthonous sheep from 19 breeds/populations from 13 geographic regions, together with previously reported 44 sequences from Chinese indigenous sheep. Phylogenetic analysis showed that all three previously defined lineages A, B, and C were found in all sampled Chinese sheep populations, except for the absence of lineage C in four populations. Network profiles revealed that the lineages B and C displayed a star-like phylogeny with the founder haplotype in the centre, and that two star-like subclades with two founder haplotypes were identified in lineage A. The pattern of genetic variation in lineage A, together with the divergence time between the two central founder haplotypes suggested that two independent domestication events have occurred in sheep lineage A. Considerable mitochondrial diversity was observed in Chinese sheep. Weak structuring was observed either among Chinese indigenous sheep populations or between Asian and European sheep and this can be attributable to long-term strong gene flow induced by historical human movements. The high levels of intra population diversity in Chinese sheep and the weak phylogeographic structuring indicated three geographically independent domestication events have occurred and the domestication place was not only confined to the Near East, but also occurred in other regions. PMID- 16704911 TI - Occupational exposure to noise, and hearing function among electro production workers. AB - OBJECTIVE: The present study has the aim to investigate the prevalence and the determinants of hearing loss among electro production workers. METHODS: Ninety three electro production workers and 51 office workers underwent audiometric test. Information regarding, socio-demographics, present and past medical history of hearing problems, use of ototoxic drugs, hobbies, and smoking habit was obtained using a self administered questionnaire. RESULTS: The levels of noise at workplace have also been measured. Electro production workers were exposed to high levels of noise. Statistical analysis has shown that 44% of electro production workers had sensorineural hearing loss located mainly at 4000 Hz. Multivariate analysis demonstrated that occupational exposure to noise appears to be the strongest predictor (odds ratio, OR: 7.51) of NIHL followed by aging (OR: 5.34), and last by smoking (OR: 2.47). CONCLUSION: Our findings indicate that electro production workers are occupationally exposed to high levels of noise, and present high rates of noise-induced hearing loss (NIHL). The observed hearing loss was located mainly at 4000 Hz. These findings underline the need for interventions to reduce the risk for the development of NIHL among electro production workers. PMID- 16704912 TI - Anatomical study of the human middle ear for the design of implantable hearing aids. AB - OBJECTIVE: To generate anatomical data on the human middle ear and adjacent structures to serve as a base for the development and optimization of new implantable hearing aid transducers. Implantable middle ear hearing aid transducers, i.e. the equivalent to the loudspeaker in conventional hearing aids, should ideally fit into the majority of adult middle ears and should utilize the limited space optimally to achieve sufficiently high maximal output levels. For several designs, more anatomical data are needed. METHODS: Twenty temporal bones of 10 formalin-fixed adult human heads were scanned by a computed tomography system (CT) using a slide thickness of 0.63 mm. Twelve landmarks were defined and 24 different distances were calculated for each temporal bone. RESULTS: A statistical description of 24 distances in the adult human middle ear which may limit or influence the design of middle ear transducers is presented. Significant inter-individual differences but no significant differences for gender, side, age or degree of pneumatization of the mastoid were found. Distances, which were not analyzed for the first time in this study, were found to be in good agreement with the results of earlier studies. CONCLUSION: A data set describing the adult human middle ear anatomy quantitatively from the point of view of designers of new implantable hearing aid transducers has been generated. In principle, the method employed in this study using standard CT scans could also be used preoperatively to rule out exclusion criteria. PMID- 16704913 TI - Severe gastrointestinal tract bleeding in a two-month-old infant due to congenital intrahepatic arterioportal fistula. AB - A 2-month-old boy was referred for assessment of severe upper gastrointestinal tract bleeding and melena. On physical examination, a continuous murmur was heard over the right upper quadrant of the abdomen. A splenomegaly and dilated veins were also noted on the abdominal wall. Liver functions were normal. There was no history of trauma or jaundice. Doppler ultrasonography, magnetic resonance arteriography and angiography suggested the presence of an intrahepatic arteriovenous fistula between the phrenic artery and the portal vein. Management consisted of successful embolization by coiling of the phrenic artery. To our knowledge this is the first documented case report of a congenital fistula between the phrenic artery and the portal vein. PMID- 16704914 TI - LC/MS/MS method for the determination of trace amounts of cefmetazole and cefpodoxime proxetil contaminants in pharmaceutical manufacturing environments. AB - In this study, a selective and sensitive LC/MS/MS method for the determination of trace amounts of cefmetazole (CMZ) and cefpodoxime proxetil (CPDXPR) contaminants in manufacturing environments was developed. The necessary sensitivity of this method was estimated based on the detection limit for Penicillin G required by the FDA and the total surface area and volume of the manufacturing facility. The detection limits of this method were estimated to be 10 pg/ml for CMZ and 5 pg/ml for CPDXPR from the signal to noise ratio and as a result satisfactory sensitivity was achieved. The method was linear in a concentration range from 0.20 to 3.20 ng/ml. The accuracy and precision were verified by the determination of the amount of CMZ and CPDXPR added to the sampling materials, a glass plate and a silica fiber filter. The mean recoveries of nine replicated determinations from the glass plate were 99.1% with 5.58%R.S.D. for CMZ and 97.1% with 3.80%R.S.D. for CPDXPR, and those from the silica fiber filter were 100.7% with 4.50%R.S.D. for CMZ and 95.4% with 2.85%R.S.D. for CPDXPR. This method has been successfully applied to the determination of CMZ and CPDXPR contaminants in samples collected from an actual manufacturing environment. PMID- 16704915 TI - Determination of honokiol and magnolol in cortex Magnoliae Officinalis by capillary electrophoresis with electrochemical detection. AB - Capillary electrophoresis with electrochemical detection has been employed for the determination of honokiol and magnolol in Cortex Magnoliae Officinalis (i.e. Magnolia Bark) for the first time. Effects of several important factors such as the concentration and the acidity of the running buffer, separation voltage, injection time, and detection potential were investigated to acquire the optimum conditions. The detection electrode was a 300 microm diameter carbon disc electrode at a working potential of +0.90 V (versus saturated calomel electrode (SCE)). The two analytes can be well separated within 6 min in a 40 cm length fused silica capillary at a separation voltage of 18 kV in a 50mM borate buffer (pH 9.2). The relation between peak current and analyte concentration was linear over about three orders of magnitude with the detection limits (S/N=3) of 0.38 and 0.51 microM for honokiol and magnolol, respectively. The proposed method has been successfully applied to monitor the two bioactive constituents in the real plant samples with satisfactory assay results. PMID- 16704916 TI - Novel therapeutic agents in ovarian cancer. AB - AIMS: Epithelial ovarian cancer is responsible for 4% of all cancer deaths in women, and the five-year overall survival of patients with advanced disease is 30 40%. Treatment currently comprises a combination of surgery and chemotherapy with carboplatin and paclitaxel. The main reason for treatment failure is that the majority of patients present with advanced disease, and current drugs are unable to effect a cure even in chemosensitive patients. This article systematically reviews novel therapeutic strategies that have been evaluated in patients with ovarian cancer in the last 5 years. METHODS: Pubmed (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?DB=pubmed) and American Society of Clinical Oncology Annual conference abstracts were searched using the terms "(phase I OR phase II OR phase III OR phase 1 OR phase 2 OR phase 3) AND (ovary OR ovarian) AND (cancer OR carcinoma)" from January 2000 to May 2005 to identify studies for potential inclusion in this review. Reviews of novel therapies in ovarian cancer were also used to identify additional clinical trials. FINDINGS: A wide range of therapeutic strategies are currently being evaluated in ovarian cancer. These include novel cytotoxics, small molecule inhibitors, monoclonal antibodies, gene therapy and immuno-therapy strategies. The rationale for the development of these agents includes enhancement of efficacy by targeting novel biological pathways, re-sensitisation to existing drugs, simplification of drug administration and/or reduction of drug-toxicity. CONCLUSIONS: Current developments have the potential to result in substantial improvements in the outlook for women with ovarian cancer. PMID- 16704917 TI - Urokinase plasminogen activator and TGF-beta production in immunosuppressed patients with and without P. Jiroveci infection. AB - Macrophages play a pivotal role in a host's defence against pulmonary infections. Macrophage functions are impaired in immunosuppressed (IS) patients, regardless of whether they are HIV-positive (HIV+) or -negative (HIV-). Several studies have indicated that urokinase plasminogen activator (uPA) and transforming growth factor beta (TGF-beta) are important factors in a host's defence against pulmonary pathogens. We measured uPA and TGF-beta activity in unstimulated peripheral blood monocytes (PBM) of both HIV-infected and non-infected IS patients with or without Pneumocystis jiroveci (formerly carinii) pneumonia (PCP). As previously found in alveolar macrophages (AMs), the majority of uPA activity was found in cell lysates. The highest values of uPA activity were found in control subjects. All the patients displayed a decreased production of uPA, irrespective of HIV infection. Similarly, active TGF-beta was higher in control subjects than in HIV+ and IS patients. The presence of P. jiroveci infection further lowered uPA and TGF-beta activity. Decreased TGF-beta activation might be a consequence of lower uPA production, which may, in turn, influence virus replication, since it has been demonstrated that TGF-beta can suppress human HIV expression in monocytes/macrophages. Further studies are warranted to elucidate whether the decrease in uPA and TGF-beta activity impairs a host's defence against P. jiroveci infection. PMID- 16704918 TI - The useful field of view test: normative data for older adults. AB - The Useful Field of View test (UFOV) is increasingly used in clinical and rehabilitation settings. To date there have been no normative data for adjusted performance comparisons across demographically-similar, elderly peers. This study examined demographic and cognitive influences on the UFOV in a sample of 2759 participants (65-94 years of age). Performance was found to differ by age and education. Regression analyses examined the relative contributions of age, education, mental status, vision, and health to UFOV performance. All of these factors were found to significantly contribute to UFOV performance, with age accounting for the most variance and education accounting for the least. Normative tables for the UFOV by age and education are provided. These norms will allow researchers and clinicians to compare UFOV performance with similar peers and may help in identifying elderly persons who would benefit from speed of processing training. PMID- 16704919 TI - Tertiary-butyl hydroperoxide induced oxidative stress and male reproductive activity in mice: role of transcription factor NF-kappaB and testicular antioxidant enzymes. AB - Reactive oxygen species (ROS) have been proposed as a major factor affecting male reproductive capacity. The present study has evaluated the possible role of oxidative stress during testicular pathogenesis in male mice exposed to tertiary butyl hydroperoxide (TBHP). TBHP was administered by daily intraperitoneal injection for 2 weeks. Treatment greatly increased lipid peroxidation in the testis and led to a significant decrease in sperm concentration and motility and a reduction in litter size relative to controls. An increase in testicular mRNA abundance of redox-regulated p50 and p65 subunits of NF-kappaB was observed after TBHP treatment. Evaluation of NF-kappaB regulated antioxidant enzymes in the testis revealed an increase in superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and glutathione-S-transferase (GST) activities and corresponding mRNA abundance. These results suggest a potential role of NF-kappaB in oxidative stress mediated changes in the physiology of male reproductive system. PMID- 16704920 TI - Th2 immune deviation induced by pregnancy: the two faces of autoimmune rheumatic diseases. AB - One of the most important immunological modifications during pregnancy is the Th1/Th2 shift, due to the progressive increase of progesterone and estrogens during pregnancy, which reach their peak-level in the third trimester of gestation. At high levels, estrogens seem mainly to suppress Th1 cytokines and stimulate Th2-mediated immunological responses as well as antibody production. For this reason Th1-mediated diseases, like rheumatoid arthritis (RA), tend to improve and Th2-mediated disease, like systemic lupus erythematosus (SLE), tend to worsen during pregnancy. SLE is the autoimmune rheumatic disease in which pregnancy most frequently occurs because it predominantly affects young females in their childbearing age. Other autoimmune rheumatic diseases, including RA, are less frequently observed during pregnancy due to their low female-to-male ratio and peak onset after the age of 40. This review is focused on the disease course, gestational outcome and management of patients with SLE and RA during pregnancy. PMID- 16704921 TI - Evaluation of minor groove binding probe and Taqman probe PCR assays: Influence of mismatches and template complexity on quantification. AB - Real-time PCR assays using 3'-minor groove binder (MGB) or Taqman probes are widely used for clinical virological testing and mutation/polymorphism detection. We compared a 3'-MGB probe to a conventional Taqman probe for linearity, sensitivity, specificity and dynamic range. The performance of the two assays was compared using plasmids containing different mismatches or using human genomic DNA as a template. Comparable linearity and sensitivity were observed for the MGB and the Taqman probe assays. Using standard conditions, none of the assays were sequence-specific. Up to five mismatches generated a detectable signal in the Taqman probe assay. The performance of the Taqman as well as the MGB probe assay was influenced by the complexity of the template, the latter, however, to a lesser degree. Overall, these results highlight the advantages of the MGB probe over the Taqman probe regarding mismatch discrimination, but suggest that optimization of reaction conditions and verification of the specificity are necessary also for MGB probes. PMID- 16704922 TI - Antifungals and renal safety--getting the balance right. AB - Many patients who receive antifungal agents are immunocompromised or critically ill. This leaves many of these patients prone to renal failure, especially following transplantation. A wealth of clinical data have shown that older antifungal agents, particularly conventional amphotericin B deoxycholate (AmBd), are still highly efficacious, except that AmBd confers extensive nephrotoxicity. This paper examines the pharmacological alternatives that could provide a more suitable balance between clinical efficacy and renal safety. PMID- 16704923 TI - Interpreting streptomycin susceptibility test results for Salmonella enterica serovar Typhimurium. AB - Resistance or susceptibility of Salmonella enterica to streptomycin is widely used as an epidemiological marker. However, there is no clear consensus on the interpretation of streptomycin susceptibility test results. Comparison of results obtained with the Clinical and Laboratory Standards Institute (CLSI) disk diffusion method, the minimum inhibitory concentration (MIC) determined by Etest and streptomycin resistance genotype for 90 isolates of S. enterica serovar Typhimurium suggests that appropriate interpretive criteria for MIC results are susceptible at or=16 mg/L. For CLSI disk diffusion, we propose susceptible at a zone diameter>or=13 mm and resistant at OATP-B>PepT1>>MDR1>MCT1 approximately MRP3 approximately BCRP approximately OCTN2>>OCT3>OCT1>OAT2. Four genes were absent: OATP8, OCT2, OAT1, and OAT3. Ranking of 11 expressed genes showed a significant correlation between human jejunum and 2-5-week-old Caco-2 cells. The expression profile in colon was, however, very different compared to both Caco-2 cells and jejunum. We conclude that the Caco-2 cells in our hands express similar transporters as the human jejunum, but are different from colon, indicating their usefulness for obtaining small intestinal transport data. In addition, we also suggest that cells with a well-defined range of culture ages should be used to minimize variability in data from experiments and even erroneous conclusions. PMID- 16704926 TI - Aqueous extract of Schizandra chinensis fruit causes endothelium-dependent and independent relaxation of isolated rat thoracic aorta. AB - An aqueous extract of Schizandra chinensis fruit (ScEx) has long been used to promote the vascular health of postmenopausal women in Korea. This study investigated the ability of ScEx to relax rat aorta constricted with norepinephrine (NE) and the mechanism(s) of such relaxation. ScEx induced partial, endothelium-dependent relaxation. In particular, the relaxation induced by lower concentrations of ScEx (0.1 and 0.3 mg/ml) was largely endothelium dependent, and was essentially abolished by NG-nitro-L-arginine, methylene blue, 1H-[1,2,3] oxadiazole [4,4-a] quinoxalin-1-one, indomethacin, or ICI 182,780. The results indicate that the response to ScEx involves enhancement of the nitric oxide (NO)-cGMP system, and that it occurs via estrogen receptors. The magnitude of the inhibition with these treatments decreased with increasing ScEx concentration, however, indicating that other vasorelaxation mechanisms are involved, which depend on the ScEx concentration. Calcium concentration-dependent contraction curves in high potassium depolarization medium were shifted significantly to the right and downward after incubation with ScEx (0.3 and 1.0 mg/ml), implying that ScEx is also involved in inhibition of the extracellular calcium influx to vascular smooth muscle. These data demonstrate that ScEx caused both endothelium-dependent and -independent vasorelaxation, which may contribute to understanding the cardiovascular protective effect of ScEx. PMID- 16704927 TI - Effects of novel leukotriene receptor antagonists in the isolated rat aorta. AB - We screened 14 novel antagonists of the LTB(4) and LTD(4) receptors (also inhibitors of LTB(4) synthesis) for their vasoactive properties in the isolated rat aorta. The compounds belong to three classes, e.g. quinoline (Q), phenetylamido (P), and resatophenone (R) derivatives. They are effectiveless in resting conditions and generally display a weak relaxing ability against contraction by either high K(+) or alpha(1) adrenoceptor activation, either in the presence or absence of a functional endothelium. There is little overlap with the generally lower concentration range where their anti-LT properties are already manifested. We could not find any correlation between any of the anti-LT properties and the vasorelaxant effects. Concerning the non-specific vasoactive properties, choice compounds of the examined groups could be further tested regarding the mechanisms of their relaxing effects. Given the many uncertainties concerning LT and vascular physiology, it may be worthy to proceed with this line of investigation. PMID- 16704928 TI - Organizing pneumonia in patients with hematologic malignancies: a steroid responsive lesion. AB - BACKGROUND: Organizing pneumonia (OP) is a distinct histopathologic lesion that occurs in a variety of clinical contexts. There have been occasional reports of OP occurring in patients with hematologic malignancies. STUDY OBJECTIVES: To examine the association of OP with hematologic malignancies and to assess the clinical course of affected patients. DESIGN: A retrospective review of our institutional experience of unexplained OP developing in patients with hematologic malignancies. SETTING: Tertiary care, referral medical center. PATIENTS: We identified 6 patients with a verified histopathologic diagnosis of OP and antecedent or concomitant diagnosis of a hematologic malignancy from the Mayo Clinic database (1995-2003). Clinical, radiologic, and outcome data were abstracted from records. RESULTS: Underlying hematologic disorders included lymphoma (2), acute leukemia (2), cutaneous T-cell lymphoma (1), and myelodysplastic syndrome (1). OP was diagnosed by surgical lung biopsy in 4 and bronchoscopic biopsy in 2. Four of the 6 patients had previously been exposed to chemotherapeutic agents, two had not. Three of the 6 patients had bone marrow transplantation prior to development of OP. Five patients were treated with prednisone and all experienced symptomatic improvement with documented radiologic resolution in 4. One patient experienced symptomatic and radiologic resolution with observation alone. Three patients ultimately died from complications of their underlying hematologic disorder and 1 patient died of unknown causes. Two patients were alive without respiratory complaints more than 1 year after lung biopsy. CONCLUSIONS: OP occurs in patients with underlying hematologic malignancies who may or may not have been treated with chemotherapy and responds favorably to corticosteroid therapy. PMID- 16704929 TI - New antibodies immobilization system into a graphite-polysulfone membrane for amperometric immunosensors. AB - Polysulfone membrane is used for the first time for the preparation of electrochemical immunosensors. A disposable immunosensor based on a porous conductor polymer graphite-polysulfone-electrode has been developed using a phase inversion technique for the determination of anti-rabbit IgG (anti-RIgG) as a model analyte. To construct the sensor, a conductor membrane was deposited on the surface of working graphite-epoxy composite (GEC) electrode. The membrane was characterized by SEM. This sensor was based on the competitive assay between free and labeled anti-RIgG for the available binding sites of immobilized rabbit IgG (RIgG). Incubation parameters were optimized in this work. The immunological reaction was detected using an enzymatic-labeling procedure (HRP enzyme) combined with the amperometric detection using H(2)O(2) as substrate and hydroquinone as mediator. This sensor shows stability during a week and a good reproducibility. The current was monitored amperometrically at -0.1 V versus SCE and this method showed a linear range of the anti-RIgG from 1 to 6 microg/ml. The detection limit was determined to be 0.77 microg/ml. PMID- 16704930 TI - The microbial diversity of inland waters. AB - The conservation and sustainable use of freshwater resources is of global importance. Microorganisms are not only the most abundant organisms in natural freshwater systems, but are also key players in ecological processes controlling water quality. Detailed knowledge of the diversity and function of microorganisms dwelling in freshwater habitats is an essential prerequisite for the sustainable management of freshwater resources. Freshwater systems are inhabited by microbial communities that are indigenous to this habitat type and usually do not occur in marine systems, saline inland waters and terrestrial habitats. Despite recent advances in the characterization of the diversity of freshwater microorganisms, knowledge essential for a holistic understanding of their ecological roles is still lacking. PMID- 16704931 TI - The Planctomycetes, Verrucomicrobia, Chlamydiae and sister phyla comprise a superphylum with biotechnological and medical relevance. AB - In the rRNA-based tree of life four bacterial phyla, comprising the Planctomycetes, Verrucomicrobia, Chlamydiae and Lentisphaerae, form together with the candidate phyla Poribacteria and OP3 a monophyletic group referred to as the PVC superphylum. This assemblage contains organisms that possess dramatically different lifestyles and which colonize sharply contrasting habitats. Some members of this group are among the most successful human pathogens, others are abundant soil microbes, and others still are of major importance for the marine nitrogen cycle and hold much promise for sustainable wastewater treatment. Recent comparative genomic and metagenomic analyses of a few representatives of this group revealed many unusual features and generated unexpected hypotheses regarding their physiology, some of which have already been confirmed experimentally. Furthermore, the availability of these genome sequences offered new insights into the evolutionary history of this peculiar group of microbes with major medical, ecological and biotechnological relevance. PMID- 16704932 TI - Homer proteins: implications for neuropsychiatric disorders. AB - Homer proteins regulate signal transduction, synaptogenesis and receptor trafficking, in addition to maintaining and regulating extracellular glutamate levels in limbo-corticostriatal brain regions. Converging preclinical observations indicate a potential role for both immediate early gene Homer isoforms and constitutively expressed Homer isoforms in behavioral pathologies associated with neuropsychiatric disorders, such as addiction and/or alcoholism, depression, anxiety, epilepsy and schizophrenia. PMID- 16704933 TI - Solutions to patient-prosthesis mismatch in aortic valve replacement. PMID- 16704934 TI - Genetic characterization of Mycobacterium tuberculosis clinical isolates with deletions in the plcA-plcB-plcC locus. AB - SETTING: The basis for Mycobacterium tuberculosis virulence is not completely understood. Analysis of the genomic structure of clinical isolates will give information that can be related to biological activities involved in virulence. OBJECTIVE: To determine the extension of the deletion in the plcA-plcB-plcC locus of selected M. tuberculosis isolates, as well as other changes in the chromosome. DESIGN: In the present work we characterized a group of M. tuberculosis isolates devoid of the plcA-plcB-plcC locus by PCR, sequencing and microarrays. RESULTS: PCR amplification of this region demonstrated a complete lack of plcA and plcB ORF's in all of the isolates. The plcC gene was completely deleted in one of the strains (DR-689) and the other three isolates still conserved part of this ORF. The loss of lateral DNA sequences ranged from 3723 to 7646bp. An IS6110 element was present in all tested strains cases, and some isolates presented the insertion of ORF's coding for proteins homologous to the ESAT-6 and QILSS families. Genomic DNA of all the strains was extracted and analyzed with an in house microarray system to observe loss of other genes possibly implicated in attenuated virulence. Two of the strains presented novel deletions; the rest of the isolates showed deletions already reported for other M. tuberculosis strains. DR-689, a Beijing type M. tuberculosis strain isolated in Canada, showed an IS6110 RFLP and a genomic deletion pattern similar to a San Francisco family of strains, although completely unrelated epidemiologically. CONCLUSION: Genomic changes in M. tuberculosis seem to occur in a controlled manner and they are possibly related to changes in its pathogenic properties. PMID- 16704935 TI - Kinetic therapy reduces complications and shortens hospital stay in patients with cardiogenic shock - a retrospective analysis. AB - BACKGROUND: Kinetic therapy (KT) has been shown to reduce complications and to shorten hospital stay in trauma patients. Data in non-surgical patients are inconclusive, and kinetic therapy has not been tested in patients with cardiogenic shock. OBJECTIVE: The present analysis compares KT with standard care in patients with cardiogenic shock. METHODS: A retrospective analysis of 133 patients with cardiogenic shock admitted to 1 academic heart center was performed. Patients with standard care (SC, turning every 2 h by the staff) were compared with kinetic therapy (KT, using oscillating air-flotation beds). MEASUREMENTS AND MAIN RESULTS: 68 patients with KT were compared with 65 patients with SC. Length of ventilator therapy was 11 days in KT and 18 days in SC (p=0.048). The mortality was comparable in both groups. Pneumonia occurred in 14 patients in KT and 39 patients in SC (p<0.001); pressure ulcers were reduced by 50% (p<0.001). Length of ICU stay (21 days in SC and 13 days in KT, p=0.009) and length of hospital stay were reduced in the patients treated with kinetic therapy. CONCLUSION: The use of KT shortens hospital stay and reduces rates of pneumonia and pressure ulcers as compared to SC. PMID- 16704936 TI - Size-exclusion chromatography of heparin oligosaccharides at high and low pressure. AB - Recent findings on specific and non-specific interactions of glycosaminoglycans (GAGs) accentuate their pivotal role in biology and the call for improved sequencing tools. The present study evaluates size-exclusion chromatography (SEC) of heparin oligosaccharides at high and low pressure, requiring amounts as low as 0.2 microgram, using conventional UV detection after depolymerization with heparin lyases. Because of their high charge at physiological pH, SEC elution volumes of heparin oligosaccharides depend on both molecular size and charge repulsion from the matrix. As a consequence, SEC elution volumes of GAGs are smaller than those of globular proteins of similar molecular weight, and this might be exploited. Accordingly, larger heparin oligosaccharides are best separated according to their size at high ionic strength of the mobile phase (>30 mM); in contrast, disaccharides are best separated according to their charge at low ionic strength, compatible with on-line coupling to mass spectrometry. Optimized SEC affords separation of characteristic heparin trisaccharides that contain uronic acid at the reducing end and suggest cellular storage of heparin as a free glycan. PMID- 16704937 TI - Intimate partner violence: prevalence, types, and chronicity in adult women. AB - BACKGROUND: Most intimate partner violence (IPV) prevalence studies do not examine the relationships between IPV types and the chronicity and severity of abuse. OBJECTIVES: Delineate prevalence, chronicity, and severity of IPV among adult women. DESIGN: Retrospective cohort study conducted by telephone survey. Data were collected in 2003 to 2005 and analyzed contemporaneously. PARTICIPANTS: English-speaking women (n=3568) aged 18 to 64 years enrolled in a U.S. health maintenance organization for 3 or more years. Response rate was 56.4%. MAIN EXPOSURE: Physical, psychological, and sexual IPV were assessed using five questions from the Behavioral Risk Factor Surveillance Survey and ten items from the Women's Experience with Battering (WEB) scale. RESULTS: Most (3429) of the respondents had at least one intimate partnership as an adult. Of these, 14.7% reported IPV of any type in the past 5 years, and 45.1% of abused women experienced more than one type. Prevalence was 7.9% in the past year, while during a woman's adult lifetime, it was 44.0%. Depending on IPV type, 10.7% to 21.0% were abused by more than one partner; duration was <1 year to 5 median years; while in 5% to 13% of the instances, IPV persisted for >20 years. IPV rates were higher for younger women, women with lower income and less education, single mothers, and those who had been abused as a child. CONCLUSIONS: The high prevalence of IPV across women's lifetimes in the previous 5 years and the previous year are documented. The present investigation provides new information of IPV chronicity, severity, and the overlap of IPV types over a woman's adult life span. PMID- 16704938 TI - Intimate partner violence and women's physical, mental, and social functioning. AB - OBJECTIVE: To describe the relationship between women's health and the timing, type, and duration of intimate partner violence (IPV) exposure. METHODS: A telephone interview was completed by 3429 women aged 18 to 64 randomly selected from a large health plan, to assess IPV exposure and heath status (response rate 56.4%). Questions from the Behavioral Risk Factor Surveillance System and the Women's Experience with Battering scale were used to construct IPV exposures: (1) recent (past 5 years) and remote (before past 5 years only) IPV exposure of any type (physical, sexual, or non-physical); (2) recent (past 5 years) IPV exposure to physical and/or sexual or non-physical only; and (3) IPV duration (0 to 2 years, 3 to 10 years, and >10 years). Health outcomes were measured using the Short Form-36 survey (SF-36), the Center for Epidemiologic Studies Depression scale, and the National Institute of Mental Health Presence of Symptoms survey. RESULTS: In adjusted models, compared to women with no IPV in their adult lifetime, more-pronounced adverse health effects were observed for women with recent (vs remote) IPV; for physical and/or sexual (vs non-physical) IPV; and for longer IPV exposure. Compared to women who never experienced IPV, women with any recent IPV (physical, sexual, or non-physical) had higher rates of severe (prevalence ratio [PR]=2.6; 95% confidence interval [CI]=1.9-3.6) and minor depressive symptoms (PR=2.3; 95% CI=1.9-2.8); higher number of physical symptoms (mean, 1.0; 95% CI=0.7-1.2); and lower SF-36 mental and social functioning scores (range, 4.3-5.5 points lower across subscales). Women with recent physical and/or sexual IPV were 2.8 times as likely to report fair/poor health, and had SF-36 scores that ranged from 5.3 to 7.8 points lower, increased risk of depressive symptoms (PR=2.6) and severe depressive symptoms (PR=4.0), and more than one additional symptom. Longer duration of IPV was associated with incrementally worse health. CONCLUSIONS: Women's health was adversely affected by the proximity, type, and duration of IPV exposure. PMID- 16704939 TI - Reactions of young adult smokers to warning labels on cigarette packages. AB - BACKGROUND: In 1984, the United States Congress enacted legislation requiring four new warning labels for cigarettes; warning labels in the United States have not changed since then. Other countries, such as Canada, have taken a more active and aggressive approach. The purpose of this study was to examine how U.S. smokers and former smokers might respond to stronger and more graphic warnings for U.S. cigarettes packages. METHODS: Data were collected in 2003 and analyzed in 2004. The perceived impact and effectiveness of the more-explicit Canadian labels was examined among young adult smokers (n=572) and former smokers (n=191) between the ages of 18 and 24 years in the United States, using a web-based survey that allowed participants to view both the text-only and the text-plus graphic warning labels. RESULTS: Both current and former smokers thought that cigarette warning labels with text plus graphics were substantially more of a deterrent than text-only labels. The perceived effectiveness of these labels was not only higher overall, but also for the specific areas of smoking-related health effects, prevention, cessation, and maintenance of abstinence. Few differences were noted by gender. CONCLUSIONS: The findings from this study support previous research that has found that text-plus-graphic warning labels are more salient and potentially more effective than text-only labels. PMID- 16704940 TI - The impact of tobacco tax cuts on smoking initiation among Canadian young adults. AB - BACKGROUND: Although the majority of smokers initiate smoking during their teenage years, significant rates of initiation occur among young adults. Adolescents are more price sensitive than adults, but little is known about the impact of tobacco taxation on smoking initiation among young adults. Using a longitudinal design, this study examined the impact of decreased cigarette price, resulting from tobacco tax cuts, on smoking initiation among Canadian young adults aged 20 to 24 years. METHODS: Using Statistics Canada's National Population Health Survey longitudinal file, this study examined young adults who did not smoke at baseline in 1994-1995 (n=636, representing over 1 million young adults) and who were reassessed at follow-up (1996-1997). Multivariable logistic regression analysis using bootstrap weights was conducted to estimate the impact of decreased cigarette price on smoking initiation. The analysis controlled for the potential confounding effect of sociodemographic and tobacco control variables. Sensitivity analyses were conducted. Price elasticity was estimated. Analyses were conducted in 2003 and 2004. RESULTS: Approximately 10% of young adults had initiated smoking at follow-up. Decreased cigarette price was significantly associated with higher smoking initiation (adjusted odds ratio per $1 decrease for a carton of cigarettes=1.15, 95% confidence interval [CI]=1.01 1.32, p=0.042). Sensitivity analyses showed similar results. Price elasticity was 3.36 (95% CI=0.07-6.75). CONCLUSIONS: Young adults are sensitive to cigarette prices. Reductions in cigarette prices will lead to increased smoking initiation among this group. Tobacco taxation should be an effective strategy to reduce smoking initiation among young adults. PMID- 16704941 TI - Differences in maternal and paternal transmission of coronary heart disease. AB - BACKGROUND: Few studies have investigated the possible differential transmission of maternal and/or paternal coronary heart disease (CHD) to offspring, after accounting for sociodemographic characteristics. METHODS: The Multigeneration Register was linked to hospital data in this study of all Swedish individuals born since 1932 and their parents. Registered cases of CHD between January 1, 1987, and December 31, 2001, were evaluated. Poisson regression was used to calculate standardized incidence ratios (SIRs) for men and women with mothers and/or fathers affected by CHD compared with men and women whose parents were not affected. All analyses were conducted in 2005. RESULTS: Maternal transmission was stronger than paternal transmission and the confidence intervals did not overlap. For women, the overall SIRs were 1.43 (95% confidence interval [CI]=1.34-1.51) for maternal transmission and 1.17 (95% CI=1.11-1.23) for paternal transmission. For men, the corresponding SIRs were 1.55 (95% CI=1.50-1.60) and 1.41 (95% CI=1.37-1.45). Even higher SIRs were found in the youngest age groups, among those with both parents affected by coronary heart disease, and among those with premature parental CHD. CONCLUSIONS: These findings might ultimately influence treatment decisions regarding risk factors and suggest the need for research focusing on genetic and intrauterine risk factors. PMID- 16704942 TI - Vascular structure and function in women: relationship with body mass index. AB - BACKGROUND: The purpose of the present study was to compare endothelial function in lean (body mass index [BMI]=18.0-24.9 kg/m2); overweight (BMI=25-29.9 kg/m2); and obese (BMI>30 kg/m2), healthy, eumenorrheic women. METHODS: Eighteen lean, 22 overweight, and 19 obese eumenorrheic middle-aged women were studied. Vascular structure and function were assessed via non-invasive ultrasound imaging of the carotid and brachial arteries. Body composition, blood pressure, fasting blood lipids, glucose, and insulin also were measured. RESULTS: The groups demonstrated significantly (p<0.001) different mean values for total body, lean body, and fat masses. The obese group demonstrated significantly (p<0.05) elevated fasting glucose and insulin levels and lower high-density lipoprotein levels as compared to the lean group. The overweight group also demonstrated elevated fasting glucose levels as compared to the lean group (p<0.05) with no significant difference from the obese group. Only systolic blood pressure differed among the three groups, being elevated in the obese group compared to the lean group (p<0.05). The obese group demonstrated significantly (p<0.05) elevated carotid artery lumen diameter, carotid artery wall cross-sectional area, and brachial artery lumen diameter with significantly (p<0.05) lower flow-mediated dilation as compared to the lean group. The overweight group demonstrated elevated carotid artery wall cross-sectional area and brachial artery lumen diameter as well as lower flow-mediated dilation as compared to the lean group (p<0.05). CONCLUSIONS: The results of this study support the hypothesis that carotid artery wall cross sectional area is elevated and flow-mediated dilation reduced in overweight and obese eumennorheic women as compared to lean counterparts in relation to BMI classification. PMID- 16704943 TI - Cancer incidence in Montana: rates for American Indians exceed those for whites. AB - OBJECTIVES: Previous studies suggested that the cancer incidence rates in American Indians and Alaska Natives were lower than in other groups. The objective of this study was to compare the cancer incidence rates in American Indians and whites in Montana. METHODS: Age-adjusted 6-year cancer incidence rates were calculated for American-Indian and white men and women in Montana to allow comparison of rates in 1991-1996 to those in 1997-2002. RESULTS: The age adjusted rates for American-Indian men were significantly higher than those for white men for all cancer sites (755+/-74 [95% confidence interval] per 100,000 vs 544+/-9 per 100,000), lung cancer (167+/-35 per 100,000 vs 83+/-4 per 100,000), and colorectal cancer (115+/-29 per 100,000 vs 61+/-4 per 100,000) from 1997 to 2002. The adjusted rates for American-Indian women were significantly higher than those for white women for all cancer sites (526+/-47 per 100,000 vs 412+/-8 per 100,000) and lung cancer (120+/-24 per 100,000 vs 56+/-3 per 100,000) during this same time period. There was a significant increase in the age-adjusted rates for all cancer sites among white men and women but not for American-Indian men or women between 1991-1996 and 1997-2002. CONCLUSIONS: There is a significant disparity in the cancer incidence rates between American Indians and whites in Montana. Regional or state-level surveillance data will be needed to describe the changing patterns of cancer incidence in many native communities in the United States. PMID- 16704944 TI - Cost effectiveness of hepatitis B vaccination at HIV counseling and testing sites. AB - BACKGROUND: Despite recent significant achievements in controlling hepatitis B virus (HBV) infection, immunizing high-risk groups against the disease remains a public health challenge in the United States. The aims of this article are to evaluate the projected cost effectiveness of hepatitis B vaccination of adults attending two major types of publicly funded HIV counseling and testing sites (CTSs)-freestanding HIV CTSs and sexually transmitted disease (STD) clinics, and to compare the cost-effectiveness of alternative vaccination and testing strategies in different subgroups in this population. METHODS: A decision model was developed to determine the economic and clinical consequences, from a societal perspective, of the following strategies in two hypothetical cohorts of 100,000 adults attending each type of site: (1) routine vaccination without screening, (2) screening for antibody to hepatitis B core antigen with an initial vaccine dose during the first visit, (3) screening and vaccination based on screening results, and (4) no intervention. Life expectancy, expected quality adjusted life years (QALYs), and medical care costs were estimated for each strategy and at each site. RESULTS: Routine vaccination was both more effective and more cost-effective than either screening strategy; under base-case assumptions, routine vaccination would cost $4400 both per QALY and per life year saved. Results for STD clinics were very similar in magnitude to those for freestanding CTSs. Results were most sensitive to clients' time and travel costs for return visits and the time-discount rate. CONCLUSIONS: Routine provision of hepatitis B vaccine at major HIV CTSs would be a highly effective and cost effective approach to preventing hepatitis B among high-risk adults in the United States. PMID- 16704945 TI - Modeling individual and physical environmental factors with adolescent physical activity. AB - BACKGROUND: Recent calls have been made to consider both environmental factors and individual-level factors in the explanation of physical activity (PA). The present study tested a conceptual model that integrated past PA, relevant environmental-level and individual-level factors and their associations with adolescent PA, using the tenets of the Theory of Planned Behavior (TPB). METHODS: Data were gathered in an adolescent sample (n=221; 60% girls) using questionnaires. PA was assessed in May 2003 and November 2003. Theory of Planned Behavior cognitions regarding PA and the environmental factors under study were assessed in November 2003. Confirmatory factor analyses and path analyses were performed using AMOS software. RESULTS: The initial structural model did not provide an acceptable fit to the data. Including a direct path from past PA to current PA significantly improved model fit to an acceptable fit. Including a direct path from past PA to environmental perceptions did not significantly improve model fit. Including a direct path from the environmental variables to current PA did not significantly improve model fit. Current PA was most strongly influenced by past PA, while environmental aesthetics and distance to PA opportunities were indirectly related to adolescents' intention to be physically active. Significant standardized path coefficients ranged from 0.14 to 0.34 and explained 17% variance in current PA. CONCLUSIONS: Combining past PA, Theory of Planned Behavior cognitions, and environmental factors increased our understanding of their relative influences on adolescent PA. Implications for future research on physical activity are discussed. PMID- 16704946 TI - Injuries in homes with certain built forms. AB - BACKGROUND: Rates of injuries may occur more frequently in different types of homes. METHODS: Retrospective population-based cohort study utilizing three linked databases: a population register, an architectural assessment of homes in the area, and an emergency department-based injury surveillance system. RESULTS: Over 58,000 homes were classified into 94 different types according to age, size, and built form. Among the 112,248 inhabitants, there were 18,044 emergency department attendances for treatment of an injury suffered in the home. Adjusted odds ratio of injuries for residents of purpose-built apartments was substantially elevated for all injuries (2.07; 95% confidence interval [CI]=1.87 2.30) and poisoning episodes (5.6; 95% CI=3.8-8.3). CONCLUSIONS: Residents of apartment buildings have substantially higher injury rates. Additional research is required to investigate the contribution of environmental hazards and behavioral factors underlying these high rates. PMID- 16704947 TI - Use and costs of nonrecommended tests during routine preventive health exams. AB - BACKGROUND: Routine office visits for preventive health exams (PHEs) are designed to identify asymptomatic diseases at early treatable stages and influence healthy behaviors. Investigation included how often diagnostic tests and procedures were performed for which evidence and guidelines recommend against in asymptomatic individuals during routine PHEs, and the costs associated with the tests and procedures. METHODS: In 2005, a cross-sectional study of office-based interventions was conducted, using National Ambulatory Medical Care Survey (NAMCS) data from 1997 to 2002. Recommendations from the United States Preventive Services Task Force (USPSTF) were used as a measure of appropriateness of diagnostic interventions during the PHE. Laboratory tests and procedures not recommended (D ranking) included urinalysis (UAs); interventions not recommended included electrocardiograms (EKGs) and x-rays. RESULTS: The frequency of ordering any of the three diagnostic interventions ranged from 5% to 37%, and at least one of the interventions was ordered 43% of the time. Annual direct costs for the three interventions range from $47 million to $194 million. CONCLUSIONS: Less use of unwarranted interventions will likely eliminate waste and improve the overall quality of health care in the United States. PMID- 16704948 TI - Preventing intimate partner violence: how we will rise to this challenge. PMID- 16704949 TI - Does a failure to count mean that it fails to count? Addressing intimate partner violence. PMID- 16704950 TI - Death and injury from motor vehicle crashes: a tale of two countries. PMID- 16704951 TI - Effects of peanut oil load on energy expenditure, body composition, lipid profile, and appetite in lean and overweight adults. AB - OBJECTIVE: We evaluated the effects of peanut oil intake on appetite, energy expenditure, body composition, and lipid profile in lean and overweight adults. METHODS: In a parallel-arm, long-term feeding trial, lean and overweight participants received a daily peanut oil load in a milk shake equivalent to 30% of their resting energy expenditure for 8 wk. Forty-eight adults (24 lean and 24 overweight, 12 male and 12 female in each group) completed the protocol. Anthropometric indices, appetite, dietary intake, energy expenditure, and plasma lipids were monitored. RESULTS: Energy intake increased significantly in the overweight but not in the lean participants. A statistically significant body weight gain (median 2.35 kg) was also observed among the overweight subjects, although this corresponded to only 43% of the theoretical weight gain. Among overweight subjects, high-density lipoprotein cholesterol increased significantly and low-density lipoprotein cholesterol decreased at week 4, but neither concentration was significantly different from baseline at week 8. Similar, but less marked, changes were observed in lean subjects. Resting energy expenditure was 5% greater (P < 0.01) in the overweight group, but no significant difference was observed in the lean subjects. No marked differences of appetite were observed over time in either group or between overweight and lean participants. CONCLUSION: These data indicate that ingestion of peanut oil elicits a weaker compensatory dietary response among overweight compared with lean individuals. Body weight increased, albeit less than theoretically predicted. The weaker effect of whole nuts on body weight reported in previous studies suggests components other than oil may be responsible. PMID- 16704952 TI - A role for fruit content in energy-restricted diets in improving antioxidant status in obese women during weight loss. AB - OBJECTIVE: The aim of the present work was to estimate the ability of two hypocaloric diets with different fruit contents to improve antioxidant biomarkers related to lipid peroxidation in obese women. METHODS: Fifteen obese women (age 32 +/- 6 y, body mass index 34.9 +/- 2.9 kg/m2) were assigned to two different dietary treatments for 8 wk. The subjects received a hypocaloric diet (600 kcal/d restriction from the measured individual energy expenditure) containing 5% (n = 8) or 15% (n = 7) energy supplied by fructose from fruits. Anthropometric measurements, blood lipid profile, plasma oxidative markers, total antioxidant capacity, and malondialdehyde (MDA) were evaluated before and after the nutritional intervention in addition to some relations among them. RESULTS: No differences in weight loss were observed between diets (5% energy from fructose in the low fruit diet -6.9 +/- 2% versus 15% energy from fructose in the high fruit diet -6.6 +/- 2%; P = 0.781). Low-density lipoprotein cholesterol levels significantly decreased (P = 0.048) in obese women who followed the high fruit diet, which was accompanied by a statistical (P = 0.046) diet-related decrease ( 30%) in the ratio of MDA to antioxidant capacity. There was a positive association between MDA diet-related change and low-density lipoprotein cholesterol (r = 0.665, P = 0.003), with antioxidant capacity directly proportional to the fiber plus fructose content associated with fruit consumption (r = 0.697, P = 0.025). CONCLUSION: A fruit-enriched hypocaloric diet appears to be more effective against oxidative stress. Consumption of antioxidant substances contained in fruit could be a useful strategy in the design of hypocaloric diets that, with the weight reduction, could increase the improvement of cardiovascular risk factors related to obesity. PMID- 16704953 TI - Associations between hypertriglyceridemia, dietary fat intake, oxidative stress, and endothelial activation in men. AB - OBJECTIVE: We examined the associations between intake of different types of dietary fat and plasma levels of oxidative stress and endothelial activation markers in men. METHODS: For that purpose, a complete physical and metabolic profile was assessed. Dietary habits of subjects were determined with a 3-d food record. We also measured fasting plasma 8-iso-prostaglandin F2alpha and oxidized low-density lipoprotein concentrations and soluble forms of vascular cell adhesion molecule-1 (sVCAM-1), intercellular adhesion molecule-1 (sICAM-1), and E selectin. All these measurements were performed with commercial enzyme-linked immunosorbent assay kits and standards. RESULTS: We found that a high total dietary fat intake was associated with high plasma sICAM-1 (r = 0.40, P < 0.005), sVCAM-1 (r = 0.31, P < 0.05), and E-selectin (r = 0.28, P < 0.05) levels. We also found that in men matched for plasma triacylglycerol levels, those consuming a diet rich in total fat (>105 g/d, n = 21) were characterized by higher circulating levels of sICAM-1 (P < 0.05) and E-selectin (P < 0.05) compared with triacylglycerol-matched individuals with a low total dietary fat intake (<105 g/d, n = 21). However, no significant difference was noted in plasma oxidized low density lipoprotein levels between groups. Further, we conducted multivariate analyses and found that saturated fatty acid intake was the only dietary variable after inclusion of other dietary variables that contributed to circulating sICAM 1 (P < 0.05) and sVCAM-1 (P < 0.05). CONCLUSION: Our study suggests that high dietary fat consumption is associated with endothelial activation in men and that this detrimental effect is likely attributable to the saturated fatty acid content of the diet. PMID- 16704954 TI - Resting energy expenditure in patients with solid tumors undergoing anticancer therapy. AB - OBJECTIVE: Few studies have investigated the resting energy expenditure (REE) of, or determined the individual predictive accuracy of prediction equations in, cancer patients undergoing anticancer therapy. This study compared the measured REE of patients with cancer undergoing anticancer therapy with (1) healthy subjects and (2) REE estimated from commonly used prediction methods. METHODS: Resting energy expenditure was measured in 18 cancer patients and 17 healthy subjects by using indirect calorimetry under standard conditions and was estimated from seven prediction methods. Fat-free mass (FFM) was measured by bioelectrical impedance analysis. Data were analyzed with regression modeling to adjust REE for FFM. Agreement between measured and predicted REE values was analyzed using the Bland-Altman approach. RESULTS: There was no significant difference in FFM-adjusted REE between cancer patients and healthy subjects (mean difference 10%). Limits of agreement were wide for all prediction methods in estimating REE as much as 40% below and up to 30% above measured REE. CONCLUSIONS: REE in cancer patients undergoing anticancer therapies does not appear to be as high as commonly thought. None of the prediction equations examined were acceptable for predicting REE of individual cancer patients or healthy subjects. PMID- 16704955 TI - Effect of fatty acids, glucose, and insulin on hepatic glucose uptake and glycolysis. AB - OBJECTIVE: There is evidence from in vitro studies that fatty acids can inhibit glucose uptake in liver. However, it is uncertain whether this happens in vivo when the liver is exposed to high levels of glucose and insulin, in combination with fatty acids, after a mixed meal. This study determined the effects of a combination of fatty acids and insulin on glucokinase (GK) activity and glycolysis in primary rat hepatocytes. METHODS: Hepatocytes were cultured with 15 mM glucose and 2 or 10 nM insulin in combination with the fatty acids palmitate, oleate, linoleate, eicosapentaenoic acid, or docosahexaenoic acid. Total GK activity and the proportion of GK in the active, unbound state were measured to determine the effect of fatty acid on the activity and cellular localization of GK. Glucose phosphorylation and glycolysis were measured in intact cells. Lactate and pyruvate synthesis and the accumulation of ketone bodies were also estimated. RESULTS: Palmitate and eicosapentaenoic acid lowered total GK activity in the presence of 2 nM insulin, but not with 10 nM insulin. In contrast, oleate, linoleate, and docosahexaenoic acid did not alter GK activity. None of the fatty acids tested inhibited glucose phosphorylation or glycolysis in intact rat hepatocytes. In addition, GK activity was unaffected by insulin concentration. CONCLUSION: Some fatty acids can act to inhibit GK activity in primary hepatocytes. However, there was no evidence that this decrease in GK activity impaired glucose phosphorylation or glycolysis. Glucose and high concentrations of insulin, which promote glucose uptake, appear to counteract any inhibitory action of fatty acids. Therefore, the presence of fatty acids in a normal mixed meal is likely to have little effect on the capacity of the liver to take up, phosphorylate, and oxidize glucose. PMID- 16704956 TI - Serum magnesium and the development of diabetes. PMID- 16704957 TI - Nutritional and metabolic abnormalities in pre-AIDS HIV infection. AB - Since the earliest reports of human immunodeficiency virus (HIV) disease, undernutrition has been associated with HIV infection, typically with the late stages of the disease (namely acquired immunodeficiency syndrome), and may advance to severe wasting and cachexia. Specific micronutrient deficiencies are also recognized to occur with HIV infection, but their actual effect on the clinical course of the disease is hard to assess. The studies reviewed provide more insight into the complex interface between undernutrition and, in some cases, obesity and HIV/acquired immunodeficiency syndrome and highlight the possibility of alleviating or curing undernutrition by means of simple and comparatively inexpensive dietary adjustments. PMID- 16704958 TI - Annexin V C/T-1 polymorphism and pregnancy complications. AB - The -1T variant of the annexin V gene, which has been suggested to have a protective role against thrombotic disease, was evaluated in 140 women with pregnancy complications and 317 control women with uncomplicated pregnancies. The presence of the -1 CT or TT genotype did not show a protective effect in normal pregnancies. PMID- 16704959 TI - Osteoporosis and osteosclerosis in sickle cell/beta-thalassemia: the role of the RANKL/osteoprotegerin axis. AB - Bone mineral density (BMD) was evaluated in 52 patients with HbS/beta thalassemia. Seventeen (32%) patients had osteopenia/osteoporosis and 30 (57%) had osteosclerosis. Bone resorption was diminished in patients with osteosclerosis and increased in those with osteopenia/osteoporosis. The sRANKL/osteoprotegerin ratio was elevated in the osteosclerotic group. Osteoporosis patients had mild renal impairment and their BMD correlated with osteoprotegerin, and bone resorption markers. Osteosclerosis patients had multiple infarctions in the studied bones that led to reduced osteoclast activity and increased BMD. In conclusion, HbS/beta-thalassemia patients may develop osteopenia/osteoporosis mainly due to marrow expansion or osteosclerosis because of ischemia after a vaso-occlusive crisis. The RANKL/ osteoprotegerin axis participates in these phenomena. PMID- 16704960 TI - The association of serum ferritin and transferrin receptor concentrations with mortality in women with human immunodeficiency virus infection. AB - BACKGROUND AND OBJECTIVES: Whether degree of iron stores influences progression of human immunodeficiency virus (HIV) disease is controversial. We studied the relationship of indirect measures of iron stores with mortality in highly active antiretroviral therapy (HAART)-naive participants from the Women's Interagency HIV Study. DESIGN AND METHODS: One hundred and fifty-eight HIV-infected women who died before July 1996 were individually matched by CD4+ cell count (within +/- 50 cells/mL) and HIV RNA level (within +/- 0.50 log10 copies/mL) to 154 controls. Serum ferritin and transferrin receptor concentrations were measured in 151 pairs of women. Results. Using multivariable conditional logistic regression models that were adjusted for self-reported antiretroviral therapy use, age, smoking status, ethnicity, hemoglobin concentration, C-reactive protein and aspartate amino transferase, a log10 increase in baseline serum ferritin concentration was associated with a 1.67-fold increase in the odds of death (95% CI: 0.98, 2.86) and a one-unit decrease in transferrin receptor to log10 ferritin ratio was associated with a 1.12-fold (95% CI: 1.01, 1.23) increase in the odds of death. INTERPRETATIONS AND CONCLUSIONS: In this study, higher indirect measures of iron status were associated with reduced survival among HAART-naive HIV-infected women. Additional prospective studies with data on direct measures of iron status along with randomized trials are needed to elucidate the current equipoise over whether iron supplementation is beneficial by preventing anemia or harmful by increasing iron stores in HIV-infected women. PMID- 16704961 TI - The effect of recombinant human intrinsic factor on the uptake of vitamin B12 in patients with evident vitamin B12 deficiency. AB - We report on the use of recombinant human intrinsic factor (rhIF) in a new vitamin B12 absorption test. Holotranscobalamin (holoTC) was measured before and 24 hours after intake of three 9-mg doses of vitamin B12 (B12) and again 24 hours after intake of the same dose of B12 together with rhIF (rhIF-B12). Nine patients with evident vitamin B12 deficiency had a significantly higher increase in holoTC after intake of rhIF-B12 than after intake of B12. Twenty-eight patients with suspected vitamin B12 deficiency showed no additional increase in holoTC after intake of rhIF-B12. We conclude that rhIF promotes B12 absorption among patients with evident vitamin B12 deficiency. PMID- 16704962 TI - The prognostic value of P-glycoprotein (ABCB) and breast cancer resistance protein (ABCG2) in adults with de novo acute myeloid leukemia with normal karyotype. AB - Multidrug resistance is a major cause of treatment failure in acute myeloid leukemia (AML). P-glycoprotein (PGP) over-expression has an unfavorable prognostic significance, while the role of breast cancer resistance protein (BCRP) is less clear, especially in AML patients with a normal karyotype. We studied 73 consecutive AML patients with a normal karyotype. BCRP was over expressed in 24 patients (33%) and was significantly co-expressed with PGP (13/24 vs 11/49, p=0.006) and with CD56. Only PGP, along with age and CD34, affected the achievement of complete remission (p=0.02), while BCRP-positive cases showed an increased risk of relapse (p=0.005) and a shorter disease-free survival (p=0.027). BCRP over-expression did not influence the achievement of remission, but significantly affected the duration of complete remissions. BCRP may, therefore, be regarded as a prognostic factor in patients with normal karyotype AML, for the design of risk-adapted post-remission therapy. PMID- 16704963 TI - The genetic basis of human erythrocyte pyridoxal kinase activity variation. AB - Thirty years ago we reported that erythrocyte pyridoxal kinase activity of African-Americans was strikingly lower than that of persons with European ancestry in a tissue-specific manner. At the time, it was impossible to elucidate the mechanism by which evolution had selectively lowered the enzyme activity in one cell type but not in others. We have now identified a promoter mutation with potential erythroid-specific properties that could be the basis of a novel mechanism of controlling cell-specific decreased activity of an essential enzyme. PMID- 16704964 TI - Detection and quantification of MBR/JH2 t(14;18) BCL-2 gene rearrangement in follicular lymphoma using a combined real-time polymerase chain reaction assay. AB - We report our experience with a new real-time polymerase chain reaction (PCR) assay applicable for simultaneous quantification and characterization of MBR/JH translocation in follicular lymphomas. This technique, which combines amplification with the FRET probe with SYBR Green I melting curve analysis, allows efficient detection of tumor cells in bone marrow or peripheral blood and their comparison with the original neoplastic clone. PMID- 16704965 TI - The frequency of HLA class I alleles in Japanese patients with bone marrow failure. AB - The frequencies of the HLA-B*4002 and HLA-A*0206 alleles in patients with aplastic anemia (AA) (n=32; 21.9%) and paroxysmal nocturanl hemoglobinuria (PNH) (n=24; 22.9%), respectively, were significantly different from those in controls (n=371; 8.6%, p<0.002 and 7.7%, p<0.001, respectively), suggesting that each specific allele in AA or PNH may be related to the immunologic pathophysiology of these disorders. PMID- 16704966 TI - Molecular cloning and characterization of rat Pomt1 and Pomt2. AB - Mammalian O-mannosylation, although an uncommon type of protein modification, is essential for normal brain and muscle development. Defective O-mannosylation causes congenital muscular dystrophy with abnormal neuronal migration [Walker Warburg syndrome (WWS)]. Here, we have identified and cloned rat Pomt1 and Pomt2, which are homologues of human POMT1 and POMT2, with identities of 86 and 90%, respectively, at the amino acid level. Coexpression of both genes was found to be necessary for enzymatic activity, as is the case with human POMT1 and POMT2. Northern blot and reverse transcriptase polymerase chain reaction (RT-PCR) analyses revealed that rat Pomt1 and Pomt2 are expressed in all tissues but most strongly in testis. In situ hybridization histochemistry of rat brain revealed that Pomt1 and Pomt2 mRNA are coexpressed in neurons (dentate gyrus and CA1-CA3 region of the hippocampus and cerebellar Purkinje cells). Two transcription initiation sites were observed in rat Pomt2, resulting in two forms: a testis form and a somatic form. The two forms had equal protein O-mannosyltransferase activity when coexpressed with rat Pomt1. Coexpression studies also showed that the human and rat protein O-mannosyltransferases are interchangeable, providing further evidence for the closeness of their structures. PMID- 16704967 TI - Protein glycosylation in Parelaphostrongylus tenuis--first description of the Galalpha1-3Gal sequence in a nematode. AB - The white-tailed deer is the definitive host of the parasitic nematode Parelaphostrongylus tenuis. This parasite also infects a wide variety of domesticated livestock, causing a debilitating neurologic disease. Glycoconjugates are becoming increasingly implicated in nematode strategies to maintain persistent infections in immunologically competent hosts. In this study, we have carried out detailed mass spectrometric analysis together with classical biochemical techniques, including western blotting and immunohistochemical staining with anticarbohydrate monoclonal antibodies and have shown that P. tenuis contains complex-type N-glycans with the antennae capped with Galalpha1 3Galbeta1-4GlcNAc sequence. By mimicking a vertebrate glycan, Galalpha1-3Gal may aid the parasite in evading immunological detection by the host. This is the first report of the Galalpha1-3Gal sequence in a nematode. PMID- 16704968 TI - Singapore's multibillion dollar gamble. AB - Figure 1Biopolis, Singapore's futuristic research hub.How does a country one fourth the size of Rhode Island with little history in biomedical science become one of the world's biomedical research giants? The answer: with a pile of money and a large dose of chutzpah. Since 2000, Singapore has dumped more than US$2 billion into developing a biomedical research industry-from scratch. Is the gamble paying off? PMID- 16704969 TI - Filling the catalytic site of cytochrome c oxidase with electrons. Reduced CuB facilitates internal electron transfer to heme a3. AB - In the reductive phase of its catalytic cycle, cytochrome c oxidase receives electrons from external electron donors. Two electrons have to be transferred into the catalytic center, composed of heme a(3) and Cu(B), before reaction with oxygen takes place. In addition, this phase of catalysis appears to be involved in proton translocation. Here, we report for the first time the kinetics of electron transfer to both heme a(3) and Cu(B) during the transition from the oxidized to the fully reduced state. The state of reduction of both heme a(3) and Cu(B) was monitored by a combination of EPR spectroscopy, the rapid freeze procedure, and the stopped-flow method. The kinetics of cytochrome c oxidase reduction by hexaamineruthenium under anaerobic conditions revealed that the rate limiting step is the initial electron transfer to the catalytic site that proceeds with apparently identical rates to both heme a(3) and Cu(B). After Cu(B) is reduced, electron transfer to oxidized heme a(3) is enhanced relative to the rate of entry of the first electron. PMID- 16704970 TI - The mammalian chitinase-like lectin, YKL-40, binds specifically to type I collagen and modulates the rate of type I collagen fibril formation. AB - YKL-40 is expressed in arthritic cartilage and produced in large amounts by cultured chondrocytes, but its exact role is unclear, and the identities of its physiological ligands remain unknown. Purification of YKL-40 from resorbing bovine nasal cartilage and chondrocyte monolayers demonstrated the existence of three isoforms, a major and minor form from resorbing cartilage and a third species from chondrocytes. Affinity chromatography experiments with purified YKL 40 demonstrated specific binding of all three forms to collagen types I, II, and III, thus identifying collagens as potential YKL-40 ligands. Binding to immobilized type I collagen was inhibited by soluble native ligand, but not heat denatured ligand, confirming a specific interaction. Binding of the chondrocyte derived species to type I collagen was also demonstrated by surface plasmon resonance analysis, and the dissociation rate constant was calculated (3.42 x 10( 3) to 4.50 x 10(-3) s(-1)). The chondrocyte-derived species was found to prevent collagenolytic cleavage of type I collagen and to stimulate the rate of type I collagen fibril formation in a concentration-dependent manner. By contrast, the cartilage major form had an inhibitory effect on type I collagen fibrillogenesis. Digestion with N-glycosidase F, endoglycosidase H and lectin blotting did not reveal any difference in the carbohydrate component of these two YKL-40 species, indicating that this does not account for the opposing effects on fibril formation rate. PMID- 16704971 TI - Cloning and characterization of mouse lung-type acyl-CoA:lysophosphatidylcholine acyltransferase 1 (LPCAT1). Expression in alveolar type II cells and possible involvement in surfactant production. AB - Phosphatidylcholine (1,2-diacyl-sn-glycero-3-phosphocholine, PC), is an important constituent of biological membranes. It is also the major component of serum lipoproteins and pulmonary surfactant. In the remodeling pathway of PC biosynthesis, 1-acyl-sn-glycero-3-phosphocholine (LPC) is converted to PC by acyl CoA:lysophosphatidylcholine acyltransferase (LPCAT, EC 2.3.1.23). Whereas LPCAT activity has been detected in several tissues, the structure and detailed biochemical information on the enzyme have not yet been reported. Here, we present the cloning and characterization of a cDNA for mouse lung-type LPCAT (LPCAT1). The cDNA encodes an enzyme of 60 kDa, with three putative transmembrane domains. When expressed in Chinese hamster ovary cells, mouse LPCAT1 exhibited Ca(2+)-independent activity with a pH optimum between 7.4 and 10. LPCAT1 demonstrated a clear preference for saturated fatty acyl-CoAs, and 1-myristoyl- or 1-palmitoyl-LPC as acyl donors and acceptors, respectively. Furthermore, the enzyme was predominantly expressed in the lung, in particular in alveolar type II cells. Thus, the enzyme might synthesize phosphatidylcholine in pulmonary surfactant and play a pivotal role in respiratory physiology. PMID- 16704972 TI - Role for an essential tyrosine in peptide amidation. AB - The catalytic core of the peptidyl-alpha-hydroxyglycine alpha-amidating lyase (PAL) domain of peptidylglycine alpha-amidating monooxygenase was investigated with respect to its ability to function as a ureidoglycolate lyase and the identity and role of its bound metal ions. The purified PAL catalytic core (PALcc) contains molar equivalents of calcium and zinc along with substoichiometric amounts of iron and functions as a ureidoglycolate lyase. Limiting iron availability in the cells synthesizing PALcc reduces the specific activity of the enzyme produced. Concentrated samples of native PALcc have an absorption maximum at 560 nm, suggestive of a phenolate-Fe(III) charge transfer complex. An essential role for a Tyr residue was confirmed by elimination of PAL activity following site-directed mutagenesis. Purified PALcc in which the only conserved Tyr residue (Tyr(654)) was mutated to Phe was secreted normally, but was catalytically inactive and lacked bound iron and bound zinc. Our data demonstrate an essential role for Tyr(654) and suggest that it serves as an Fe(III) ligand in an essential iron-zinc bimetallic site. PMID- 16704974 TI - Surface expression of ASIC2 inhibits the amiloride-sensitive current and migration of glioma cells. AB - Gliomas are primary brain tumors with a complex biology characterized by antigenic and genomic heterogeneity and a propensity for invasion into normal brain tissue. High grade glioma cells possess a voltage-independent, amiloride inhibitable, inward Na+ current. This current does not exist in normal astrocytes or low grade tumor cells. Inhibition of this conductance decreases glioma growth and cell migration making it a potential therapeutic target. Our previous results have shown that the acid-sensing ion channels (ASICs), members of the epithelial Na+ channel (ENaC)/degenerin (DEG) family of ion channels are part of this current pathway. We hypothesized that one member of the ENaC/DEG family, ASIC2, is retained intracellularly and that it is the lack of functional expression of ASIC2 at the cell surface that results in hyperactivity of this conductance in high grade gliomas. In this study we show that the chemical chaperone, glycerol, and the transcriptional regulator, sodium 4-phenylbutyrate, inhibit the constitutively activated inward current and reduce cell growth and migration in glioblastoma multiforme. The results suggest that these compounds induce the movement of ASIC2 to the plasma membrane, and once there, the basally active inward current characteristic of glioma cells is abolished by inherent negative regulatory mechanisms. This in turn compromises the ability of the glioma cell to migrate and proliferate. These results support the hypothesis that the conductance pathway in high grade glioma cells is comprised of ENaC/DEG subunits and that abolishing this channel activity promotes a reversion of a high grade glioma cell to a phenotype resembling that of normal astrocytes. PMID- 16704975 TI - Multiple importins function as nuclear transport receptors for the Rev protein of human immunodeficiency virus type 1. AB - The Rev protein of human immunodeficiency virus type 1 is an RNA-binding protein that is required for nuclear export of unspliced and partially spliced viral mRNAs. Nuclear import of human immunodeficiency virus type 1 Rev has been suggested to depend on the classic nuclear transport receptor importin beta, but not on the adapter protein importin alpha. We now show that, similar to importin alpha, Rev is able to dissociate RanGTP from recycling importin beta, a reaction that leads to the formation of a novel import complex. Besides importin beta, the transport receptors transportin, importin 5, and importin 7 specifically interact with Rev and promote its nuclear import in digitonin-permeabilized cells. A single arginine-rich nuclear localization sequence of Rev is required for interaction with all importins tested so far. In contrast to the importin beta binding domain of importin alpha, Rev interacts with an N-terminal fragment of importin beta. Transportin contains two independent binding sites for Rev. Hence, the mode of interaction of importin beta and transportin with Rev is clearly distinct from that with their classic import cargoes. Taken together, the viral protein takes advantage of multiple cellular transport pathways for its nuclear accumulation. PMID- 16704973 TI - An outer membrane enzyme encoded by Salmonella typhimurium lpxR that removes the 3'-acyloxyacyl moiety of lipid A. AB - The Salmonella and related bacteria modify the structure of the lipid A portion of their lipopolysaccharide in response to environmental stimuli. Some lipid A modifications are required for virulence and resistance to cationic antimicrobial peptides. We now demonstrate that membranes of Salmonella typhimurium contain a novel hydrolase that removes the 3'-acyloxyacyl residue of lipid A in the presence of 5 mM Ca2+. We have identified the gene encoding the S. typhimurium lipid A 3'-O-deacylase, designated lpxR, by screening an ordered S. typhimurium genomic DNA library, harbored in Escherichia coli K-12, for expression of Ca2+ dependent 3'-O-deacylase activity in membranes. LpxR is synthesized with an N terminal type I signal peptide and is localized to the outer membrane. Mass spectrometry was used to confirm the position of lipid A deacylation in vitro and the release of the intact 3'-acyloxyacyl group. Heterologous expression of lpxR in the E. coli K-12 W3110, which lacks lpxR, resulted in production of significant amounts of 3'-O-deacylated lipid A in growing cultures. Orthologues of LpxR are present in the genomes of E. coli O157:H7, Yersinia enterocolitica, Helicobacter pylori, and Vibrio cholerae. The function of LpxR is unknown, but it could play a role in pathogenesis because it might modulate the cytokine response of an infected animal. PMID- 16704976 TI - Inhibitory role of Ser-425 of the alpha1 2.2 subunit in the enhancement of Cav 2.2 currents by phorbol-12-myristate, 13-acetate. AB - Voltage-gated calcium channels (Ca(v)) 2.2 currents are potentiated by phorbol-12 myristate, 13-acetate (PMA), whereas Ca(v) 2.3 currents are increased by both PMA and acetyl-beta-methylcholine (MCh). MCh-selective sites were identified in the alpha(1) 2.3 subunit, whereas the identified PMA sites responded to both PMA and MCh (Kamatchi, G. L., Franke, R., Lynch, C., III, and Sando, J. J. (2004) J. Biol. Chem. 279, 4102-4109; Fang, H., Franke, R., Patanavanich, S., Lalvani, A., Powell, N. K., Sando, J. J., and Kamatchi, G. L. (2005) J. Biol. Chem. 280, 23559 23565). The hypothesis that PMA sites in the alpha(1) 2.2 subunit are homologous to the PMA-responsive sites in alpha(1) 2.3 subunit was tested with Ser/Thr --> Ala mutations in the alpha(1) 2.2 subunit. WT alpha(1) 2.2 or mutants were expressed in Xenopus oocytes in combination with beta1b and alpha2/delta subunits. Inward current (I(Ba)) was recorded using Ba(2+) as the charge carrier. T422A, S1757A, S2108A, or S2132A decreased the PMA response. In contrast, S425A increased the response to PMA, and thus, it was considered an inhibitory site. Replacement of each of the identified stimulatory Ser/Thr sites with Asp increased the basal current and decreased the PMA-induced enhancement, consistent with regulation by phosphorylation at these sites. Multiple mutant combinations showed (i) greater inhibition than that caused by the single Ala mutations; (ii) that enhancement observed when Thr-422 and Ser-2108 are available may be inhibited by the presence of Ser-425; and (iii) that the combination of Thr-422, Ser-2108, and either Ser-1757 or Ser-2132 can provide a greater response to PMA when Ser-425 is replaced with Ala. The homologous sites in alpha(1) 2.2 and alpha(1) 2.3 subunits seem to be functionally different. The existence of an inhibitory phosphorylation site in the I-II linker seems to be unique to the alpha(1) 2.2 subunit. PMID- 16704977 TI - Modular structure of solubilized human apolipoprotein B-100. Low resolution model revealed by small angle neutron scattering. AB - Being intimately involved in cholesterol transport and lipid metabolism human low density lipoprotein (LDL) plays a prominent role in atherogenesis and cardiovascular diseases. The receptor-mediated cellular uptake of LDL is triggered by apolipoprotein B-100 (apoB-100), which represents the single protein moiety of LDL. Due to the size and hydrophobic nature of apoB-100, its structure is not well characterized. Here we present a low resolution structure of solubilized apoB-100. We have used small angle neutron scattering in combination with advanced shape reconstruction algorithms to generate a three-dimensional model of lipid-free apoB-100. Our model clearly reveals that apoB-100 is composed of distinct domains connected by flexible regions. The apoB-100 molecule adopts a curved shape with a central cavity. In comparison to LDL-associated apoB-100, the lipid-free protein is expanded, whereas according to spectroscopic data the secondary structure is widely preserved. Finally, the low resolution model was used as a template to reconstruct a hypothetical domain organization of apoB-100 on LDL, including information derived from a secondary structure prediction. PMID- 16704978 TI - Binding to Rab3A-interacting molecule RIM regulates the presynaptic recruitment of Munc13-1 and ubMunc13-2. AB - Transmitter release at synapses between nerve cells is spatially restricted to active zones, where synaptic vesicle docking, priming, and Ca2+-dependent fusion take place in a temporally highly coordinated manner. Munc13s are essential for priming synaptic vesicles to a fusion competent state, and their specific active zone localization contributes to the active zone restriction of transmitter release and the speed of excitation-secretion coupling. However, the molecular mechanism of the active zone recruitment of Munc13s is not known. We show here that the active zone recruitment of Munc13 isoforms Munc13-1 and ubMunc13-2 is regulated by their binding to the Rab3A-interacting molecule RIM1alpha, a key determinant of long term potentiation of synaptic transmission at mossy fiber synapses in the hippocampus. We identify a single point mutation in Munc13-1 and ubMunc13-2 (I121N) that, depending on the type of assay used, strongly perturbs or abolishes RIM1alpha binding in vitro and in cultured fibroblasts, and we demonstrate that RIM1alpha binding-deficient ubMunc13-2(I121) is not efficiently recruited to synapses. Moreover, the levels of Munc13-1 and ubMunc13-2 levels are decreased in RIM1alpha-deficient brain, and Munc13-1 is not properly enriched at active zones of mossy fiber terminals of the mouse hippocampus if RIM1alpha is absent. We conclude that one function of the Munc13/RIM1alpha interaction is the active zone recruitment of Munc13-1 and ubMunc13-2. PMID- 16704979 TI - Structural determinants that target the hepatitis C virus core protein to lipid droplets. AB - Hepatitis C virus core protein is targeted to lipid droplets, which serve as intracellular storage organelles, by its C-terminal domain, termed D2. From circular dichroism and nuclear magnetic resonance analyses, we demonstrate that the major structural elements within D2 consist of two amphipathic alpha-helices (Helix I and Helix II) separated by a hydrophobic loop. Both helices require a hydrophobic environment for folding, indicating that lipid interactions contribute to their structural integrity. Mutational studies revealed that a combination of Helix I, the hydrophobic loop, and Helix II is essential for efficient lipid droplet association and pointed to an in-plane membrane interaction of the two helices at the phospholipid layer interface. Aside from lipid droplet association, membrane interaction of D2 is necessary for folding and stability of core following maturation at the endoplasmic reticulum membrane by signal peptide peptidase. These studies identify critical determinants within a targeting domain that enable trafficking and attachment of a viral protein to lipid droplets. They also serve as a unique model for elucidating the specificity of protein-lipid interactions between two membrane-bound organelles. PMID- 16704980 TI - Biochemical and structural analysis of Helix pomatia agglutinin. A hexameric lectin with a novel fold. AB - Helix pomatia agglutinin (HPA) is a N-acetylgalactosamine (GalNAc) binding lectin found in the albumen gland of the roman snail. As a constituent of perivitelline fluid, HPA protects fertilized eggs from bacteria and is part of the innate immunity system of the snail. The peptide sequence deduced from gene cloning demonstrates that HPA belongs to a family of carbohydrate-binding proteins recently identified in several invertebrates. This domain is also present in discoidin from the slime mold Dictyostelium discoideum. Investigation of the lectin specificity was performed with the use of glycan arrays, demonstrating that several GalNAc-containing oligosaccharides are bound and rationalizing the use of this lectin as a cancer marker. Titration microcalorimetry performed on the interaction between HPA and GalNAc indicates an affinity in the 10(-4) M range with an enthalpy-driven binding mechanism. The crystal structure of HPA demonstrates the occurrence of a new beta-sandwich lectin fold. The hexameric quaternary state was never observed previously for a lectin. The high resolution structure complex of HPA with GalNAc characterizes a new carbohydrate binding site and rationalizes the observed preference for alphaGalNAc-containing oligosaccharides. PMID- 16704981 TI - Genetic basis for the synthesis of the immunomodulatory mannose caps of lipoarabinomannan in Mycobacterium tuberculosis. AB - Lipoarabinomannan (LAM) is a high molecular weight, heterogenous lipoglycan present in abundant quantities in Mycobacterium tuberculosis and many other actinomycetes. In M. tuberculosis, the non-reducing arabinan termini of the LAM are capped with alpha1-->2 mannose residues; in some other species, the arabinan of LAM is not capped or is capped with inositol phosphate. The nature and extent of this capping plays an important role in disease pathogenesis. MT1671 in M. tuberculosis CDC1551 was identified as a glycosyltransferase that could be involved in LAM capping. To determine the function of this protein a mutant strain of M. tuberculosis CDC1551 was studied, in which MT1671 was disrupted by transposition. SDS-PAGE analysis showed that the LAM of the mutant strain migrated more rapidly than that of the wild type and did not react with concanavalin A as did wild-type LAM. Structural analysis using NMR, gas chromatography/mass spectrometry, endoarabinanase digestion, Dionex high pH anion exchange chromatography, and matrix-assisted laser desorption ionization-time-of flight mass spectrometry demonstrated that the LAM of the mutant strain was devoid of mannose capping. Since an ortholog of MT1671 is not present in Mycobacterium smegmatis mc(2)155, a recombinant strain was constructed that expressed this protein. Analysis revealed that the LAM of the recombinant strain was larger than that of the wild type, had gained concanavalin A reactivity, and that the arabinan termini were capped with a single mannose residue. Thus, MT1671 is the mannosyltransferase involved in deposition of the first of the mannose residues on the non-reducing arabinan termini and the basis of much of the interaction between the tubercle bacillus and the host cell. PMID- 16704982 TI - Localization of the phosphoethanolamine methyltransferase of the human malaria parasite Plasmodium falciparum to the Golgi apparatus. AB - Phosphatidylcholine is the most abundant phospholipid in the membranes of Plasmodium falciparum, the agent of severe human malaria. The synthesis of this phospholipid occurs via two routes, the CDP-choline pathway, which uses host choline as a precursor, and the plant-like serine decarboxylase phosphoethanolamine methyltransferase (SDPM) pathway, which uses host serine as a precursor. Although various components of these pathways have been identified, their cellular locations remain unknown. We have previously reported the identification and characterization of the phosphoethanolamine methyltransferase, Pfpmt, of P. falciparum and shown that it plays a critical role in the synthesis of phosphatidylcholine via the SDPM pathway. Here we provide the first evidence that the transmethylation step of the SDPM pathway occurs in the parasite Golgi apparatus. We show that the level of Pfpmt protein in the infected erythrocyte is regulated in a stage-specific fashion, with high levels detected during the trophozoite stage at the peak of parasite membrane biogenesis. Confocal microscopy revealed that Pfpmt is not cytoplasmic. Immunoelectron microscopy revealed that Pfpmt localizes to membrane structures that extend from the nuclear membrane but that it only partially co-localizes with the endoplasmic reticulum marker BiP. Using transgenic parasites expressing green fluorescent protein targeted to different cellular compartments, a complete co-localization was detected with Rab6, a marker of the Golgi apparatus. Together these studies provide the first evidence that the transmethylation step of the SDPM pathway of P. falciparum occurs in the Golgi apparatus and indicate an important role for this organelle in parasite membrane biogenesis. PMID- 16704983 TI - Ethanolaminephosphate side chain added to glycosylphosphatidylinositol (GPI) anchor by mcd4p is required for ceramide remodeling and forward transport of GPI proteins from endoplasmic reticulum to Golgi. AB - Glycosylphosphatidylinositol (GPI) anchors of mammals as well as yeast contain ethanolaminephosphate side chains on the alpha1-4- and the alpha1-6-linked mannoses of the anchor core structure (protein-CO-NH-(CH(2))(2)-PO(4)-6Manalpha1 2Manalpha1-6Manalpha1-4GlcNH(2)-inositol-PO(4)-lipid). In yeast, the ethanolaminephosphate on the alpha1-4-linked mannose is added during the biosynthesis of the GPI lipid by Mcd4p. MCD4 is essential because Gpi10p, the mannosyltransferase adding the subsequent alpha1-2-linked mannose, requires substrates with an ethanolaminephosphate on the alpha1-4-linked mannose. The Gpi10p ortholog of Trypanosoma brucei has no such requirement. Here we show that the overexpression of this ortholog rescues mcd4Delta cells. Phenotypic analysis of the rescued mcd4Delta cells leads to the conclusion that the ethanolaminephosphate on the alpha1-4-linked mannose, beyond being an essential determinant for Gpi10p, is necessary for an efficient recognition of GPI lipids and GPI proteins by the GPI transamidase for the efficient transport of GPI anchored proteins from the endoplasmic reticulum to Golgi and for the physiological incorporation of ceramides into GPI anchors by lipid remodeling. Furthermore, mcd4Delta cells have a marked defect in axial bud site selection, whereas this process is normal in gpi7Delta and gpi1. This also suggests that axial bud site selection specifically depends on the presence of the ethanolaminephosphate on the alpha1-4-linked mannose. PMID- 16704984 TI - Structure and catalytic mechanism of a novel N-succinyl-L-ornithine transcarbamylase in arginine biosynthesis of Bacteroides fragilis. AB - A Bacteroides fragilis gene (argF'(bf)), the disruption of which renders the bacterium auxotrophic for arginine, was expressed and its recombinant protein purified and studied. The novel protein catalyzes the carbamylation of N-succinyl L-ornithine but not L-ornithine or N-acetyl-L-ornithine, forming N-succinyl-L citrulline. Crystal structures of this novel transcarbamylase complexed with carbamyl phosphate and N-succinyl-L-norvaline, as well as sulfate and N-succinyl L-norvaline have been determined and refined to 2.9 and 2.8 A resolution, respectively. They provide structural evidence that this protein is a novel N succinyl-L-ornithine transcarbamylase. The data provided herein suggest that B. fragilis uses N-succinyl-L-ornithine rather than N-acetyl-L-ornithine for de novo arginine biosynthesis and therefore that this pathway in Bacteroides is different from the canonical arginine biosynthetic pathway of most organisms. Comparison of the structures of the new protein with those recently reported for N-acetyl-L ornithine transcarbamylase indicates that amino acid residue 90 (B. fragilis numbering) plays an important role in conferring substrate specificity for N succinyl-L-ornithine versus N-acetyl-L-ornithine. Movement of the 120 loop upon substrate binding occurs in N-succinyl-L-ornithine transcarbamylase, while movement of the 80 loop and significant domain closure take place as in other transcarbamylases. These findings provide new information on the putative role of succinylated intermediates in arginine biosynthesis and on the evolution of transcarbamylases. PMID- 16704985 TI - Associations of genetic variants in the estrogen receptor coactivators PPARGC1A, PPARGC1B and EP300 with familial breast cancer. AB - The mitogen effect of the ovarian steroid estrogen is a strong risk factor for breast cancer development. This effect is mainly mediated by the estrogen receptor alpha, a hormone inducible transcription factor, which activates gene expression through recruiting multiple coactivators, such as PPARGC1A, PPARGC1B and EP300. We tested the hypothesis that non-conservative, putative functional amino acid exchanges in PPARGC1A, PPARGC1B and EP300 act as low-penetrance familial breast cancer risk factors. The analysis of 816 BRCA1/2 mutation negative familial breast cancer patients and 1012 controls revealed an association of the PPARGC1A Thr612Met polymorphism with familial breast cancer (OR = 1.35, 95% CI 1.00-1.81, P = 0.049), high-risk familial breast cancer (OR = 1.51, 95% CI 1.08-2.12, P = 0.017) and bilateral familial breast cancer (OR = 2.30, 95% CI 1.24-4.28, P = 0.009). Logistic regression analyses of the PPARGC1B Ala203Pro variant showed an increased familial breast cancer risk of heterozygous and homozygous variant allele carriers (OR = 1.48, 95% CI 1.15-1.91, P = 0.002). The genotype-combination analysis of the associated PPARGC1A Thr612Met variant and the associated PPARGC1B Ala203Pro variant suggests an allele dose-dependent breast cancer risk (P(trend) = 0.0004). Our results indicate for the first time the importance of inherited variants in the estrogen receptor coactivator genes PPARGC1A and PPARGC1B for familial breast cancer susceptibility. Owing to their impact on estrogen signaling, these polymorphisms might also influence adjuvant anti-estrogen therapy, using agents such as tamoxifen and raloxifen, and outcome of breast cancer patients. PMID- 16704986 TI - Brassica vegetable consumption reduces urinary F2-isoprostane levels independent of micronutrient intake. AB - Isothiocyanates and indoles (e.g. indole-3-carbinol) from Brassica vegetables (e.g. broccoli) induce Phase I and Phase II enzymes responsible for the oxidation, reduction and metabolism of endogenous and exogenous carcinogens. Brassica vegetables also contain micronutrients that may provide additional DNA protection from reactive oxygen species. This randomized crossover trial (n = 20) compares the effects of a Brassica Vegetable (BV) intervention against a Micronutrient and Fiber Supplementation (M+F) intervention on urinary F2 isoprostane levels (F2-iP), a stable biomarker of systemic oxidative stress. Brassica intake was monitored by repeated 24 h recalls, urinary ITC levels and questionnaire. Urinary F2-iP levels were measured by mass spectrometry from first morning urine samples collected at Baseline and after each intervention, and change in natural log transformed urinary F2-iP levels were analyzed using repeated measures regression. Brassica consumption increased from 2 grams/day (g/d) during the Baseline or M+F intervention periods to 218 g/d during the BV intervention, whereas exposure to most antioxidant vitamins and minerals was greatest during the M+F intervention. F2-iP levels significantly decreased by 22.0 or 21.8% during the BV intervention compared with Baseline or the M+F intervention (P = 0.05, P = 0.05, respectively). Urinary F2-iP levels did not significantly differ between Baseline and the M+F intervention (difference = 0.2%; P = 0.98). Brassica intake has been associated with reduced risk of colon, lung, bladder, breast, prostate and other cancers. Our results suggest that Brassica consumption reduces systemic oxidative stress independent of the vitamin and mineral content of these vegetables. PMID- 16704987 TI - Arachidonic acid-induced gene expression in colon cancer cells. AB - It is well documented that arachidonic acid (AA) and its metabolites are intimately linked to cancer biology. However, the downstream mechanism(s) that link AA levels to cancer cell proliferation remain to be elucidated. Initial experiments in the current study showed that exogenous AA and inhibitors of AA metabolism that lead to the accumulation of unesterified AA are cytotoxic to the colon cancer cell line, HCT-116. Additionally, exogenous AA and triacsin C, an inhibitor of AA acylation, induced apoptosis and related caspase-3 activity in a transcriptionally dependent manner. Gene array analysis revealed that both exogenous AA and triacsin C alter the expression of similar genes in HCT-116 cells. For example, both downregulate several genes with well-documented roles in cell survival and apoptotic resistance. Conversely, both upregulate genes encoding activator protein-1 (AP-1) transcription factors, which have known roles in inducing apoptosis, and genes that counteract ras (Erk/MAPK) growth signaling pathways. Real-time polymerase chain reaction and immunoblotting demonstrated that mRNA and protein levels of one of the major AP-1 transcription factors, c Jun, is markedly elevated by exogenous AA and triacsin C. Additionally, the cyclooxygenase inhibitor, sulindac sulfide, increases c-Jun mRNA levels. Together, these studies reveal that the generation of intracellular AA and its subsequent impact on gene expression probably represents a critical step that regulates colon cancer cell proliferation. PMID- 16704988 TI - Repression of androgen receptor in prostate cancer cells by phenethyl isothiocyanate. AB - BACKGROUND: Prostate cancer usually progresses to androgen refractory after an initial anti-androgen treatment. The androgen receptor (AR) is a pivotal factor for the androgen-mediated growth and maintenance of the prostate. Abnormality of the AR, such as overexpression has been postulated to be related to the hormone independent growth of the cancer. Although we previously demonstrated that the AR expression could be modulated by isothiocyanates, which are natural constituents of cruciferous vegetables, the mechanism, however, remained to be clarified. We have since investigated the mechanism of phenethyl isothiocyanate (PEITC) in AR regulation. METHODS: A human androgen dependent prostate cancer cell line LNCaP (AD) and its sub-line LNCaP (AI), i.e. androgen independent but overexpressing AR, were exposed to PEITC. The effects of PEITC on cell growth and AR expression/transcription were analyzed with MTT assay, real-time PCR and western blotting. The AR promoter activity was analyzed with the reporter activity after transfection with pAR-luc. The effects on Sp1, the major transcription factor of the AR, were tested with Sp1-luc activity, western blotting and electrophoretic mobility shift assay. RESULTS: PEITC induced a significant growth inhibition, with equal IC(50), in both AD and AI cells. The AR present in both cells was repressed as demonstrated with real-time PCR and western blot. PEITC mediates dual effects at transcriptional and post-translational levels to regulate the AR. At transcriptional level the AR level was reduced via inhibition of the transcription factor Sp1, and at post-translational level by accelerating protein degradation. CONCLUSION: PEITC represses AR transcription and expression, and mediates growth arrest in androgen dependent and independent prostate cancer cells. With the AR modulation and growth attenuation, PEITC and possibly other isothiocyanates, may prevent and inhibit hormone sensitive and refractory prostate cancer. PMID- 16704989 TI - Dual role of beta-carotene in combination with cigarette smoke aqueous extract on the formation of mutagenic lipid peroxidation products in lung membranes: dependence on pO2. AB - Results from some intervention trials indicated that supplemental beta-carotene enhanced lung cancer incidence and mortality in chronic smokers. The aim of this study was to verify the hypothesis that high concentrations of the carotenoid, under the pO2 present in lung (100-150 mmHg), may exert deleterious effects through a prooxidant mechanism. To test this hypothesis, we examined the interactions of beta-carotene and cigarette smoke condensate (tar) on the formation of lipid peroxidation products in rat lung microsomal membranes enriched in vitro with varying beta-carotene concentrations (from 1 to 10 nmol/mg prot) and then incubated with tar (6-25 microg/ml) under different pO2. As markers of lipid peroxidation, we evaluated the levels of conjugated dienes and malondialdehyde, possessing mutagenic and pro-carcinogenic activity. The exposure of microsomal membranes to tar induced a dose-dependent enhancement of lipid peroxidation, which progressively increased as a function of pO2. Under a low pO2 (15 mmHg), beta-carotene acted clearly as an antioxidant, inhibiting tar-induced lipid peroxidation. However, the carotenoid progressively lost its antioxidant efficiency by increasing pO2 (50-100 mmHg) and acted as a prooxidant at pO2 ranging from 100 to 760 mmHg in a dose-dependent manner. Consistent with this finding, the addition of alpha-tocopherol (25 microM) prevented the prooxidant effects of the carotenoid. beta-Carotene auto-oxidation, measured as formation of 5,6-epoxy-beta,beta-carotene, was faster at high than at low pO2 and the carotenoid was more rapidly consumed in the presence of tar. These data point out that the carotenoid may enhance cigarette smoke-induced oxidative stress and exert potential deleterious effects at the pO2 normally present in lung tissue. PMID- 16704990 TI - Indole-3-carbinol in the maternal diet provides chemoprotection for the fetus against transplacental carcinogenesis by the polycyclic aromatic hydrocarbon dibenzo[a,l]pyrene. AB - The fetus and neonate are sensitive targets for chemically induced carcinogenesis. Few studies have examined the risk/benefit of chemoprotective phytochemicals, given in the maternal diet, against transplacental carcinogenesis. In this study, B6129 SF1/J (AHR(b-1/d)) and 129Sv/ImJ (AHR(d/d)) mice were cross-bred. The polycyclic aromatic hydrocarbon, dibenzo[a,l]pyrene (DBP), was administered to pregnant mice (15 mg/kg, gavage) on gestation day 17, and 2000 p.p.m. indole-3-carbinol (I3C), a chemoprotective phytochemical from cruciferous vegetables, was fed to half of the mice from gestation day 9 until weaning. Offspring born to dams fed I3C exhibited markedly fewer mortalities (P < 0.0001). Maternal dietary exposure to I3C also significantly lowered lung tumor multiplicity (P = 0.035) in offspring surviving to 10 months of age. The I3C chemoprotection was independent of either maternal or fetal AHR genotype. The bioavailability of DBP to fetal target tissue was demonstrated by assessing DNA covalent adduction with a (33)P-post-labeling assay. The bioavailability of I3C was determined by dosing a subset of pregnant mice with [(14)C]-I3C. Addition of chemoprotective agents to the maternal diet during pregnancy and nursing may be an effective new approach in reducing the incidence of cancers in children and young adults. PMID- 16704991 TI - The function of melanotransferrin: a role in melanoma cell proliferation and tumorigenesis. AB - Melanotransferrin (MTf) or melanoma tumor antigen p97 is an iron (Fe) binding transferrin homolog expressed highly on melanomas and at lower levels on normal tissues. It has been suggested that MTf is involved in a variety of processes such as Fe metabolism and cellular differentiation. Considering the crucial role of Fe in many metabolic pathways, for example, DNA synthesis, it is important to understand the function of MTf. To define the roles of MTf, two models were developed: (i) an MTf knockout (MTf-/-) mouse and (ii) downregulation of MTf expression in melanoma cells by post-transcriptional gene silencing (PTGS). Examination of the MTf-/- mice demonstrated no differences compared with wild type littermates. However, microarray analysis showed differential expression of molecules involved in proliferation such as Mef2a, Tcf4, Gls and Apod in MTf-/- mice compared with MTf+/+ littermates. Considering the role of MTf in melanoma cells, PTGS was used to downregulate MTf mRNA and protein levels by >90 and >80%, respectively. This resulted in inhibition of proliferation and migration. As found in MTf-/- mice, in melanoma cells with suppressed MTf expression, hMEF2A and hTCF4 were upregulated compared with parental cells. Furthermore, when melanoma cells with decreased MTf expression were injected into nude mice, tumor growth was markedly reduced, suggesting a role for MTf in proliferation and tumorigenesis. PMID- 16704992 TI - Science for life: a conversation with Nobel laureate David Baltimore. Interview by Barbara J Culliton. AB - As a man with equal interests in science and science policy, David Baltimore has been at the forefront of many of the important debates that have shaped science since the 1970s. Very much engaged in the initial discussions about the use of recombinant DNA technology, Baltimore had a front-row seat as the biotechnology industry developed. He was also a major player in the decision that resulted in funding of the Human Genome Project by the National Institutes of Health (NIH). Baltimore discusses biotechnology, science education, and the need for a strong dialogue among scientists and scholars in the health policy community. PMID- 16704993 TI - Comparative genomic hybridization analysis of human oocytes and polar bodies. AB - BACKGROUND: Classical cytogenetic methods and fluorescent in situ hybridization (FISH) have been employed for the analysis of chromosomal abnormalities in human oocytes. However, these methods are limited by the need to spread the sample on a microscope slide, a process that risks artefactual chromosome loss. Comparative genomic hybridization (CGH) is a DNA-based method that enables the investigation of the entire chromosome complement. We optimized and evaluated a CGH protocol for the chromosomal analysis of first polar bodies (PBs) and oocytes. The protocol was then employed to obtain a detailed picture of meiosis I errors in human oogenesis. METHODS: 107 MII oocyte-PB complexes were examined using whole genome amplification (WGA) and CGH. RESULTS: Data was obtained for 100 complexes, donated from 46 patients of average age 32.5 (range 18-42). 22 complexes from 15 patients were abnormal, giving an aneuploidy rate of 22%. CONCLUSIONS: The results presented in this study more than double the quantity of CGH data from female gametes currently available. Abnormalities caused by whole chromosome non disjunction, unbalanced chromatid predivision and chromosome breakage were reliably identified using the CGH protocol. Analysis of the data revealed a preferential participation of chromosome X and the smaller autosomes in aneuploidy and provided further evidence for the existence of age-independent factors in female aneuploidy. PMID- 16704994 TI - The type of catheter has no impact on the pregnancy rate after intrauterine insemination: a randomized study. AB - BACKGROUND: This study was done to test the hypothesis that intrauterine insemination (IUI) using a soft-tip catheter results in a higher live birth rate than IUI using a hard-tip catheter. METHODS: Five hundred and forty patients were randomized into those inseminated with a soft-tip catheter (group 1, n = 267) and those inseminated with a hard-tip catheter (group 2, n = 269). Four patients were excluded. Main outcome measures included pregnancy rate and live birth rate per cycle. RESULTS: Both groups were similar with regard to female age, duration of infertility, ovarian stimulation and sperm quality. No significant differences were observed between group 1 and group 2 regarding clinical pregnancy rate per cycle (20 versus 19%), live birth rate per cycle (15 versus 14%), multiple live birth rate per cycle (4 versus 6%) and multiple live birth per total of live births (5 versus 8%, overall 6%), respectively. CONCLUSION: Our hypothesis that IUI using a soft tip catheter results in a higher live birth rate per cycle than IUI using a hard-tip catheter was not confirmed in this study. Multiple live birth rate after treatment with low-dose gonadotrophins and IUI can be kept low (6%). PMID- 16704995 TI - Comparison between a GnRH antagonist and a GnRH agonist flare-up protocol in oocyte donors: a randomized clinical trial. AB - BACKGROUND: Little information is available on the outcome of controlled ovarian hyperstimulation (COH) using GnRH antagonist in oocyte donation cycles especially in comparison with the short GnRH agonist protocol. This study was aimed at comparing the two stimulation protocols in oocyte donation (OD) cycles. METHODS: A total of 113 donors randomly received COH using either GnRH antagonist or GnRH agonist. The primary endpoint was the mean number of mature oocytes retrieved per started donor cycle. Secondary endpoints were the mean number of cumulus-oocyte complexes (COCs) retrieved, the mean proportion of mature oocytes, pregnancy and implantation rates in recipients. RESULTS: Oocytes were distributed to 166 recipients. The mean number (+/- SD) of COC (11.6 +/- 5.8 versus 12.1 +/- 6.7), mature oocytes (8.4 +/- 4.4 versus 8.9 +/- 5.3) and the proportion of mature oocytes (70.8 versus 75.7%) retrieved per started donor cycle were similar in the antagonist and agonist groups, respectively. The implantation rate (26.1 versus 30.1%), clinical (40.2 versus 45.6%) and ongoing pregnancy rate per recipient cycle (32.2 versus 37.9%) were comparable in antagonist and agonist protocols, respectively. CONCLUSIONS: Similar mean number of mature oocytes and comparable pregnancy rates are achieved after OD in which donors received COH using GnRH antagonist or short GnRH agonist protocols. PMID- 16704996 TI - The photoprotective role of epidermal anthocyanins and surface pubescence in young leaves of grapevine (Vitis vinifera). AB - BACKGROUND AND AIMS: Depending on cultivar, surfaces of young leaves of Vitis vinifera may be glabrous-green ('Soultanina') or transiently have anthocyanins ('Siriki') or pubescence ('Athiri'). A test is made of the hypothesis that anthocyanins and pubescence act as light screens affording a photoprotective advantage to the corresponding leaves, and an assessment is made of the magnitude of their effect. METHODS: Measurements were made on young leaves of the three cultivars in spring under field conditions. Photosynthetic gas-exchange and in vivo chlorophyll fluorescence were measured. Photosynthetic and photoprotective pigments were analysed by HPLC. KEY RESULTS: Compared with glabrous-green leaves, both anthocyanic and pubescent leaves had greater dark-adapted PSII photochemical efficiency and net photosynthesis. In leaves possessing either anthocyanins or pubescence, the ratio of xanthophyll cycle components to total chlorophyll, and mid-day de-epoxidation state of the xanthophyll cycle were considerably smaller, than in glabrous-green leaves. These differences were more evident in pubescent leaves, probably indicating that trichomes were more effective in decreasing light stress than anthocyanins in the epidermis. CONCLUSIONS: Light screens, especially in the form of pubescence, decrease the risk of photoinhibition whilst allowing leaves to maintain a smaller content of xanthophyll cycle components and depend less on xanthophyll cycle energy dissipation. This combination of photoprotective features, i.e. decreased photon flux to the photosynthetic apparatus and lower xanthophyll cycle utilization rates may be particularly advantageous under stressful conditions. PMID- 16704997 TI - Functional characterization of phosphoenolpyruvate carboxykinase-type C4 leaf anatomy: immuno-, cytochemical and ultrastructural analyses. AB - BACKGROUND AND AIMS: Species having C4 photosynthesis belonging to the phosphoenolpyruvate carboxykinase (PEP-CK) subtype, which are found only in family Poaceae, have the most complex biochemistry among the three C4 subtypes. In this study, biochemical (western blots and immunolocalization of some key photosynthetic enzymes) and structural analyses were made on several species to further understand the PEP-CK system. This included PEP-CK-type C4 species Urochloa texana (subfamily Panicoideae), Spartina alterniflora and S. anglica (subfamily Chloridoideae), and an NADP-ME-type C4 species, Echinochloa frumentacea, which has substantial levels of PEP-CK. KEY RESULTS: Urochloa texana has typical Kranz anatomy with granal chloroplasts scattered around the cytoplasm in bundle sheath (BS) cells, while the Spartina spp. have BS forming long adaxial extensions above the vascular tissue and with chloroplasts in a strictly centrifugal position. Despite some structural and size differences, in all three PEP-CK species the chloroplasts in mesophyll and BS cells have a similar granal index (% appressed thylakoids). Immunolocalization studies show PEP-CK (which catalyses ATP-dependent decarboxylation) is located in the cytosol, and NAD-ME in the mitochondria, in BS cells, and in the BS extensions of Spartina. In the NADP ME species E. frumentacea, PEP-CK is also located in the cytosol of BS cells, NAD ME is very low, and the source of ATP to support PEP-CK is not established. CONCLUSIONS: Representative PEP-CK species from two subfamilies of polyphyletic origin have very similar biochemistry, compartmentation and chloroplast grana structure. Based on the results with PEP-CK species, schemes are presented with mesophyll and BS chloroplasts providing equivalent reductive power which show bioenergetics of carbon assimilation involving C4 cycles (PEP-CK and NAD-ME, the latter functioning to generate ATP to support the PEP-CK reaction), and the consequences of any photorespiration. PMID- 16704998 TI - Phylogeny and classification of Paris (Melanthiaceae) inferred from DNA sequence data. AB - BACKGROUND AND AIMS: Paris (Melanthiaceae) is a temperate genus of about 24 perennial herbaceous species distributed from Europe to eastern Asia. The delimitation of the genus and its subdivisions are unresolved questions in the taxonomy of Paris. The objective of this study is to test the generic and infrageneric circumscription of Paris with DNA sequence data. METHODS: Phylogenetic analysis of 21 species of Paris based on nuclear ITS and plastid psbA-trnH and trnL-trnF DNA sequence data, alone and in combination, was employed to assess previous classifications. KEY RESULTS: Paris is monophyletic in all analyses. Neither of the two traditionally recognized subgenera (Paris and Daiswa) are monophyletic. Sections Axiparis, Kinugasa, Paris and Thibeticae are monophyletic in only some of the analyses. Species of sections Dunnianae, Fargesianae and Marmoratae are consistently intercalated among species of section Euthyra in all analyses. Strong discordance between nuclear and plastid lineages is detected. CONCLUSIONS: The data support the classification of Paris as a single genus rather than as three genera (Daiswa, Kinugasa and Paris sensu stricto). They provide justification for the transfer of section Axiparis from subgenus Paris to subgenus Daiswa and for the combination of sections Dunnianae, Fargesianae and Marmoratae into section Euthyra. The nuclear-plastid discordance is interpreted as the result of interspecific hybridization among sympatric species. PMID- 16704999 TI - Exine micromorphology of Orchidinae (Orchidoideae, Orchidaceae): phylogenetic constraints or ecological influences? AB - BACKGROUND AND AIMS: Pollen characters have been widely used in defining evolutionary trends in orchids. In recent years, information on pollination biology and phylogenetic patterns within Orchidinae has become available. Hence, the aim of the presented work is to re-evaluate exine micromorphology of Orchidinae in light of recent phylogenetic studies and to test whether pollen micromorphology strictly depends on phylogenetic relationships among species or whether it is influenced by the marked differences in pollination ecology also reported among closely related species. METHODS: Pollen sculpturing of 45 species of Orchidinae and related taxa was investigated using scanning electron microscopy. To cover potential intraspecific variation, several accessions of the same species were examined. KEY RESULTS: Orchidinae show remarkable variation in exine sculpturing, with a different level of variation within species groups. In some genera, such as Serapias (rugulate) and Ophrys (psilate to verrucate), intrageneric uniformity corresponds well to a common pollination strategy and close relationships among species. However, little exine variability (psilate scabrate and scabrate-rugulate) was also found in the genus Anacamptis in spite of striking differences in floral architecture and pollination strategies. A larger variety of exine conditions was found in genera Dactylorhiza (psilate, psilate-scabrate and reticulate) and Orchis s.s. (psilate, reticulate, perforate rugulate and baculate) where no unequivocal correspondence can be found to either phylogenetic patterns or pollination strategies. CONCLUSIONS: Changes in pollen characteristics do not consistently reflect shifts in pollination strategy. A unique trend of exine evolution within Orchidinae is difficult to trace. However, the clades comprising Anacamptis, Neotinea, Ophrys and Serapias show psilate to rugulate or scabrate pollen, while that of the clade comprising Chamorchis, Dactylorhiza, Gymnadenia, Orchis s.s., Platanthera, Pseudorchis and Traunsteinera ranges from psilate to reticulate. Comparison of the data with exine micromorphology from members of the tribe Orchidieae and related tribes suggests a possible general trend from reticulate to psilate. PMID- 16705000 TI - Nutrient availability and atmospheric CO2 partial pressure modulate the effects of nutrient heterogeneity on the size structure of populations in grassland species. AB - BACKGROUND AND AIMS: Size-asymmetric competition occurs when larger plants have a disproportionate advantage in competition with smaller plants. It has been hypothesized that nutrient heterogeneity may promote it. Experiments testing this hypothesis are inconclusive, and in most cases have evaluated the effects of nutrient heterogeneity separately from other environmental factors. The aim of this study was to test, using populations of Lolium perenne, Plantago lanceolata and Holcus lanatus, two hypotheses: (a) nutrient heterogeneity promotes size asymmetric competition; and (b) nutrient heterogeneity interacts with both atmospheric CO2 partial pressure (P(CO2)) and nutrient availability to determine the magnitude of this response. METHODS: Microcosms consisting of monocultures of the three species were grown for 90 d in a factorial experiment with the following treatments: P(CO2) (37.5 and 70 Pa) and nutrient availability (NA; 40 and 120 mg of N added as organic material) combined with different spatial distribution of the organic material (NH; homogeneous and heterogeneous). Differences in the size of individual plants within populations (size inequality) were quantified using the coefficient of variation of individual above-ground biomass and the combined biomass of the two largest individuals in each microcosm. Increases in size inequality were associated with size-asymmetric competition. KEY RESULTS: Size inequality increased when the nutrients were heterogeneously supplied in the three species. The effects of NH on this response were more pronounced under high nutrient supply in both Plantago and Holcus (significant NA x NH interactions) and under elevated P(CO2) in Plantago (significant P(CO2) x NA x NH interaction). No significant two- and three-way interactions were found for Lolium. CONCLUSIONS: Our first hypothesis was supported by our results, as nutrient heterogeneity promoted size-asymmetric competition in the three species evaluated. Nutrient supply and P(CO2) modified the magnitude of this effect in Plantago and Holcus, but not in Lolium. Thus, our second hypothesis was partially supported. PMID- 16705001 TI - Euro Heart Survey on adult congenital heart disease: concern for the complexity of care. PMID- 16705002 TI - The impact of guideline compliant medical therapy on clinical outcome in patients with stable angina: findings from the Euro Heart Survey of stable angina. AB - AIMS: The European Society of Cardiology published guidelines for the management of stable angina in 1997, with the objective of promoting an evidence-based approach to the condition. This study focuses on the impact of guideline compliant medical treatment on clinical outcome in patients with stable angina. METHODS AND RESULTS: The Euro Heart Survey of Stable Angina is a multicentre prospective observational study conducted between 2002 and 2003. Patients with a clinical diagnosis of stable angina by a cardiologist were enrolled and follow-up was conducted at 1 year. The primary outcome of interest was death or myocardial infarction (MI). The increasing intensity of guideline compliant medical therapy was quantified by means of a simple treatment score based on the use of guideline advocated therapies: antiplatelets, statins, and beta-blockers. A total of 3779 patients were included in the initial survey. Increasing intensity of guideline compliant therapy at initial assessment was associated with a reduction in death and MI during follow-up in patients with angina and confirmed coronary disease (HR 0.68; 95% CI 0.49-0.95 per unit increase in treatment score). All cardiovascular events were also significantly reduced in this subgroup (HR 0.82; 95% CI 0.69-0.97). The benefits of guideline compliant therapy were only observed in patients with objective evidence of coronary disease. CONCLUSION: Guideline compliant medical therapy improves clinical outcome in patients with stable angina and objective evidence of coronary disease. PMID- 16705003 TI - The effect of steroids on the clinical course and outcome of neonates with meconium aspiration syndrome. AB - Meconium Aspiration Syndrome (MAS) is an important cause of neonatal morbidity and mortality. The present study was undertaken to evaluate the role of steroids in the management of MAS. This was a double blinded randomized controlled trial and a prospective Interventional Study over one-year period in the neonatal unit of the Lady Hardinge Medical College and associated Kalawati Saran Children's hospital. Fifty-one babies of MAS which were randomly distributed into three groups, Control, systemic and nebulized steroids. Methyl prednisolone was given i.v. in dose of 0.5 mg/kg/day in two divided doses. Budecort was given by nebulization in dose of 50 microgram 12 hourly. Infants were assessed in terms of duration of stay, oxygen dependence, X-ray clearances and also assessed for short term adverse effects. There was a statistically significant difference in the duration of stay, duration of oxygen dependence and radiological clearance. The use of steroids was not associated with an increased incidence of sepsis. The conclusion is that steroids alter the course of Meconium Aspiration Syndrome and favorably affect the outcome. PMID- 16705004 TI - Impact of specialty of admitting physician and type of hospital on care and outcome for myocardial infarction in England and Wales during 2004-5: observational study. AB - OBJECTIVE: To examine process of care and outcome for patients admitted with acute myocardial infarction to hospitals in England and Wales in relation to type of consultant care and type of hospital. DESIGN: Observational study of 88,782 patients admitted with myocardial infarction during 2004-5, using records from the national audit of myocardial infarction project (MINAP) database. OUTCOME MEASURES: Use of reperfusion treatment and secondary prevention drugs, use of angiography, and 90 day mortality of patients admitted under the care of cardiologists and non-cardiologists in hospitals with and without facilities for coronary intervention. FINDINGS: 36% of patients were admitted under the care of a cardiologist and 20% to a hospital with coronary interventional facilities. Patients admitted under cardiologists had fewer comorbidities than other patients and were more likely to have reperfusion treatment (12,266/14,433 (85%) v 13,682/17,064 (80%)) and appropriate secondary prevention drugs. Overall, 27,431/79,374 (35%) of patients had angiography. Relatively more patients admitted to interventional hospitals (8167/14,661; 56%) than to other hospitals had angiography (19,264/64,713; 30%). The adjusted risk of death by 90 days for patients treated in interventional compared with non-interventional hospitals was 0.93 (95% confidence interval 0.82 to 1.06). The adjusted risk of death at 90 days for patients admitted under cardiologists compared with non-cardiologists was 0.86 (0.81 to 0.91). CONCLUSIONS: Patients cared for by cardiologists had less comorbidity than other patients. They were more likely to receive proved treatments and angiography, and they had a lower adjusted 90 day mortality. Large differences existed in the use of angiography between interventional and non interventional hospitals. These findings show wide variations in the management and outcome of patients with myocardial infarction in England and Wales. PMID- 16705005 TI - Neonatal nephrocalcinosis: long term follow up. AB - AIMS: To assess the spontaneous resolution of neonatal nephrocalcinosis and its long term effects on renal function. METHODS: Fourteen very low birthweight preterm babies with nephrocalcinosis were followed up at 5-7 years of age; 14 controls were matched for sex, gestation, and birth weight. Height, weight, blood pressure, and renal symptomatology were recorded, and a renal ultrasound scan was performed. Early morning urine osmolality and creatinine ratios of albumin, phosphate, calcium, oxalate and beta microglobulin were determined. Urea and electrolytes in the study group were determined, and glomerular filtration rate (GFR) and TmP/GFR (tubular reabsorption of phosphate per GFR) were calculated. Statistical analysis was performed on a group basis using the Mann-Whitney confidence interval. RESULTS: Mean age was 6.9 years (range 5.81-7.68). An early morning urine osmolality >700 mOsm/kg was achieved in all cases. In two cases and four controls, the calcium/creatinine ratio was >0.7 mmol/mmol. In all cases, the GFR was normal (median 132.6 ml/min/1.73 m(2) (range 104.1-173.1)). Median TmP/GFR was 1.22 mmol/l (0.73-1.61), with two having levels below the normal range. These did not have persisting nephrocalcinosis. Nephrocalcinosis was found in three of the 12 cases scanned and one control. There were no significant differences in urine biochemistry. CONCLUSIONS: Resolution of nephrocalcinosis occurred in 75% of cases. No evidence was found to suggest that nephrocalcinosis is associated with renal dysfunction in the long term. There was evidence of hypercalciuria in the cases and controls, suggesting that prematurity may be a risk factor. PMID- 16705007 TI - Palliative care for prenatally diagnosed lethal fetal abnormality. AB - Diagnosis of lethal fetal abnormality raises challenging decisions for parents and clinicians. Most parents opt for termination, which may include feticide. Advances in imaging seem unlikely to lead to earlier diagnoses. Perinatal palliative care offers an alternative. Parental decision making and the clinical aspects of perinatal palliative care were studied after a prenatal diagnosis of lethal fetal abnormality in 20 pregnancies. 40% of parents chose to continue the pregnancy and pursue perinatal palliative care. Six of these eight babies were liveborn and lived for between 1(1/2) h and 3 weeks. PMID- 16705006 TI - Intrauterine neutrophil activation is associated with pulmonary haemorrhage in preterm infants. AB - BACKGROUND: Clinical and experimental studies showing lung damage associated with biologically active neutrophil derivatives suggests the possibility that intrauterine neutrophil activation may predispose to the development of pulmonary haemorrhage in extremely low birthweight infants early after birth. OBJECTIVES: To assess neutrophil functional activity in cord blood from extremely low birthweight infants who subsequently developed severe pulmonary haemorrhage. METHODS: Neutrophil functional activity was evaluated in the cord blood of preterm neonates (gestational age <28 weeks and weight <800 g) who developed pulmonary haemorrhage (n = 6) and in controls who did not (n = 6). Infants with congenital abnormalities and noteworthy maternal complications such as diabetes, pre-eclampsia, haemorrhagic disorders, and connective tissue disease were excluded. Neutrophils isolated from cord blood immediately after delivery were tested to evaluate neutrophil chemotaxis, random motility, and chemiluminescence. Standard methodology was used. RESULTS: Chemotaxis and chemiluminescence of neutrophils from cord blood of infants with pulmonary haemorrhage was higher than in the controls (34.8 (2.0) v 28.6 (0.8) microm, p<0.001, and 95.8 (8.5) v 73.2 (3.6) cpm x10(3), p<0.001 respectively). Random motility of cord blood neutrophils was comparable in the two groups (28.6 (1.2) v 25.3 (1.2) microm, p<0.09). CONCLUSIONS: Intrauterine induction of chemotactic activity and metabolic oxygenation of neutrophils may predispose to the development of pulmonary haemorrhage in extremely low birthweight infants with respiratory distress early after birth. PMID- 16705008 TI - Preschool healthcare utilisation related to home oxygen status. AB - OBJECTIVE: To determine, in prematurely born children who had bronchopulmonary dysplasia (BPD), if respiratory morbidity, healthcare utilisation, and cost of care during the preschool years were influenced by use of supplementary oxygen at home after discharge from the neonatal intensive care unit. DESIGN: Observational study. SETTING: Four tertiary neonatal intensive care units. PATIENTS: 190 children, median gestational age 27 weeks (range 22-31), 70 of whom received supplementary oxygen when discharged home. INTERVENTIONS: Review of hospital and general practitioner records together with a parent completed respiratory questionnaire. MAIN OUTCOME MEASURES: Healthcare utilisation, cost of care, cough, wheeze, and use of an inhaler. RESULTS: Seventy children had supplementary oxygen at home (home oxygen group), but only one had a continuous requirement for home oxygen beyond 2 years of age. There were no significant differences in the gestational age or birth weight of the home oxygen group compared with the rest of the cohort. However, between 2 and 4 years of age inclusive, the home oxygen group had more outpatient attendances (p = 0.0021) and specialist attendances (p = 0.0023), and, for respiratory problems, required more prescriptions (p<0.0001). Their total cost of care was higher (p<0.0001). In addition, more of the home oxygen group wheezed more than once a week (p = 0.0486) and were more likely to use an inhaler (p<0.0001). CONCLUSIONS: Children with BPD who have supplementary oxygen at home after discharge have increased respiratory morbidity and healthcare utilisation in the preschool years. PMID- 16705009 TI - Characterization of high-light-responsive promoters of the psaAB genes in Synechocystis sp. PCC 6803. AB - In cyanobacteria, transcription of genes encoding subunits of PSI is tightly repressed under high-light conditions. To elucidate the molecular mechanism, we examined the promoter architecture of the psaAB genes encoding reaction center subunits of PSI in a cyanobacterium Synechocystis sp. PCC 6803. Primer extension analysis showed the existence of two promoters, P1 and P2, both of which are responsible for the light intensity-dependent transcription of the psaAB genes. Deletion analysis of the upstream region of psaAB fused to bacterial luciferase reporter genes (luxAB) indicated that the light response of these promoters is achieved in a totally different manner. The cis-element required for the light response of P1, designated as PE1, was located just upstream of the -35 element of P1 and was comprised of AT-rich sequence showing significant homology to the upstream promoter (UP)-element often found in strong bacterial promoters. PE1 activated P1 under low-light conditions, and the down-regulation of P1 was achieved by rapid inactivation of PE1 upon the shift to high-light conditions. On the other hand, the cis-element required for the light response of P2, designated as HNE2, was located upstream of the P1 region, far from the basal promoter of P2. The down-regulation of P2 seemed to be attained through the negative regulation by HNE2 activated only under high-light conditions. DNA gel mobility shift assays showed that at least five regions in psaAB promoters were responsible for the binding of putative regulatory protein factors. PMID- 16705010 TI - Endosperm-limited Brassicaceae seed germination: abscisic acid inhibits embryo induced endosperm weakening of Lepidium sativum (cress) and endosperm rupture of cress and Arabidopsis thaliana. AB - The endosperm is a barrier for radicle protrusion of many angiosperm seeds. Rupture of the testa (seed coat) and rupture of the endosperm are two sequential events during the germination of Lepidium sativum L. and Arabidopsis thaliana (L.) Heyhn. Abscisic acid (ABA) specifically inhibits the endosperm rupture of these two closely related Brassicaceae species. Lepidium seeds are large enough to allow the direct measurement of endosperm weakening by the puncture force method. We found that the endosperm weakens prior to endosperm rupture and that ABA delays the onset and decreases the rate of this weakening process in a dose dependent manner. An early embryo signal is required and sufficient to induce endosperm weakening, which afterwards appears to be an organ-autonomous process. Gibberellins can replace this embryo signal; de novo gibberellin biosynthesis occurs in the endosperm and weakening is regulated by the gibberellin/ABA ratio. Our results suggest that the control of radicle protrusion during the germination of Brassicaceae seeds is mediated, at least in part, by endosperm weakening. We propose that Lepidium is an emerging Brassicaceae model system for endosperm weakening and that the complementary advantages of Lepidium and Arabidopsis can be used in parallel experiments to investigate the molecular mechanisms of endosperm weakening. PMID- 16705011 TI - Developing a move-set for protein model refinement. AB - MOTIVATION: A wide variety of methods for the construction of an atomic model for a given amino acid sequence are known, the more accurate being those that use experimentally determined structures as templates. However, far fewer methods are aimed at refining these models. The approach presented here carefully blends models created by several different means, in an attempt to combine the good quality regions from each into a final, more refined, model. RESULTS: We describe here a number of refinement operators (collectively, 'move-set') that enable a relatively large region of conformational space to be searched. This is used within a genetic algorithm that reshuffles and repacks structural components. The utility of the move-set is demonstrated by introducing a cost function, containing both physical and other components guiding the input structures towards the target structure. We show that our move-set has the potential to improve the conformation of models and that this improvement can be beyond even the best template for some comparative modelling targets. AVAILABILITY: The populus software package and the source code are available at http://bmm.cancerresearchuk.org/~offman01/populus.html. PMID- 16705012 TI - FCP: functional coverage of the proteome by structures. AB - MOTIVATION: Tools and resources for translating the remarkable growth witnessed in recent years in the number of protein structures determined experimentally into actual gain in the functional coverage of the proteome are becoming increasingly necessary. We introduce FCP, a publicly accessible web tool dedicated to analyzing the current state and trends of the population of structures within protein families. FCP offers both graphical and quantitative data on the degree of functional coverage of enzymes and nuclear receptors by existing structures, as well as on the bias observed in the distribution of structures along their respective functional classification schemes. AVAILABILITY: http://cgl.imim.es/fcp CONTACT: jmestres@imim.es. PMID- 16705013 TI - Protein classification using probabilistic chain graphs and the Gene Ontology structure. AB - MOTIVATION: Probabilistic graphical models have been developed in the past for the task of protein classification. In many cases, classifications obtained from the Gene Ontology have been used to validate these models. In this work we directly incorporate the structure of the Gene Ontology into the graphical representation for protein classification. We present a method in which each protein is represented by a replicate of the Gene Ontology structure, effectively modeling each protein in its own 'annotation space'. Proteins are also connected to one another according to different measures of functional similarity, after which belief propagation is run to make predictions at all ontology terms. RESULTS: The proposed method was evaluated on a set of 4879 proteins from the Saccharomyces Genome Database whose interactions were also recorded in the GRID project. Results indicate that direct utilization of the Gene Ontology improves predictive ability, outperforming traditional models that do not take advantage of dependencies among functional terms. Average increase in accuracy (precision) of positive and negative term predictions of 27.8% (2.0%) over three different similarity measures and three subontologies was observed. AVAILABILITY: C/C++/Perl implementation is available from authors upon request. PMID- 16705014 TI - Distance-based clustering of CGH data. AB - MOTIVATION: We consider the problem of clustering a population of Comparative Genomic Hybridization (CGH) data samples. The goal is to develop a systematic way of placing patients with similar CGH imbalance profiles into the same cluster. Our expectation is that patients with the same cancer types will generally belong to the same cluster as their underlying CGH profiles will be similar. RESULTS: We focus on distance-based clustering strategies. We do this in two steps. (1) Distances of all pairs of CGH samples are computed. (2) CGH samples are clustered based on this distance. We develop three pairwise distance/similarity measures, namely raw, cosine and sim. Raw measure disregards correlation between contiguous genomic intervals. It compares the aberrations in each genomic interval separately. The remaining measures assume that consecutive genomic intervals may be correlated. Cosine maps pairs of CGH samples into vectors in a high dimensional space and measures the angle between them. Sim measures the number of independent common aberrations. We test our distance/similarity measures on three well known clustering algorithms, bottom-up, top-down and k-means with and without centroid shrinking. Our results show that sim consistently performs better than the remaining measures. This indicates that the correlation of neighboring genomic intervals should be considered in the structural analysis of CGH datasets. The combination of sim with top-down clustering emerged as the best approach. AVAILABILITY: All software developed in this article and all the datasets are available from the authors upon request. CONTACT: juliu@cise.ufl.edu. PMID- 16705015 TI - Combining multiple microarrays in the presence of controlling variables. AB - MOTIVATION: Microarray technology enables the monitoring of expression levels for thousands of genes simultaneously. When the magnitude of the experiment increases, it becomes common to use the same type of microarrays from different laboratories or hospitals. Thus, it is important to analyze microarray data together to derive a combined conclusion after accounting for the differences. One of the main objectives of the microarray experiment is to identify differentially expressed genes among the different experimental groups. The analysis of variance (ANOVA) model has been commonly used to detect differentially expressed genes after accounting for the sources of variation commonly observed in the microarray experiment. RESULTS: We extended the usual ANOVA model to account for an additional variability resulting from many confounding variables such as the effect of different hospitals. The proposed model is a two-stage ANOVA model. The first stage is the adjustment for the effects of no interests. The second stage is the detection of differentially expressed genes among the experimental groups using the residuals obtained from the first stage. Based on these residuals, we propose a permutation test to detect the differentially expressed genes. The proposed model is illustrated using the data from 133 microarrays collected at three different hospitals. The proposed approach is more flexible to use, and it is easier to accommodate the individual covariates in this model than using the meta-analysis approach. AVAILABILITY: A set of programs written in R will be electronically sent upon request. PMID- 16705016 TI - Symptom reporting in childhood asthma: a comparison of assessment methods. AB - BACKGROUND: One barrier to receiving adequate asthma care is inaccurate estimations of symptom severity. AIMS: To interview parents of children with asthma in order to: (1) describe the range of reported illness severity using three unstructured methods of assessment; (2) determine which assessment method is least likely to result in a "critical error" that could adversely influence the child's care; and (3) determine whether the likelihood of making a "critical error" varies by sociodemographic characteristics. METHODS: A total of 228 parents of children with asthma participated. Clinical status was evaluated using structured questions reflecting National Asthma Education and Prevention Panel (NAEPP) criteria. Unstructured assessments of severity were determined using a visual analogue scale (VAS), a categorical assessment of severity, and a Likert scale assessment of asthma control. A "critical error" was defined as a parent report of symptoms in the lower 50th centile for each method of assessment for children with moderate-severe persistent symptoms by NAEPP criteria. RESULTS: Children with higher severity according to NAEPP criteria were rated on each unstructured assessment as more symptomatic compared to those with less severe symptoms. However, among the children with moderate-severe persistent symptoms, many parents made a critical error and rated children in the lower 50th centile using the VAS (41%), the categorical assessment (45%), and the control assessment (67%). The likelihood of parents making a critical error did not vary by sociodemographic characteristics. CONCLUSIONS: All of the unstructured assessment methods tested yielded underestimations of severity that could adversely influence treatment decisions. Specific symptom questions are needed for accurate severity assessments. PMID- 16705017 TI - Improved outcome of referrals for intestinal transplantation in the UK. AB - AIM: To describe the outcome of children with intestinal failure referred to Birmingham Children's Hospital (BCH) for consideration of intestinal transplantation (ITx), to determine factors for an adverse outcome and to analyse the impact of post-1998 strategies on survival. SUBJECTS AND METHODS: A retrospective analysis was performed of children referred for ITx assessment from January 1989 to December 2003. Children were assessed by a multidisciplinary team and categorised into: (a) stable on parenteral nutrition; (b) unsuitable for transplantation (Tx); and (c) recommended for Tx. To analyse the impact of the post-1998 strategies on survival, a comparison was made between the two eras (pre 1998 and post-1998). RESULTS: 152 children with chronic intestinal failure were identified (63M:89F, median age 10 months (range 1-170)). After assessment, 69 children were considered stable on parenteral nutrition (5-year survival 95%); 28 children were unsuitable for Tx (5-year survival 4%); and 55 children were recommended for Tx (5-year survival 35%, which includes 14 children who died waiting for size-matched organs). Twenty three ITx and nine isolated liver transplants (iLTx) were performed. In a multivariate analysis, the following factors in combination had an adverse effect on survival: the presence of a primary mucosal disorder (p = 0.007, OR ratio 3.16, 95% CI 1.37 to 7.31); absence of involvement of a nutritional care team at the referring hospital (p = 0.001, OR ratio 2.55, 95% CI 1.44 to 4.52); and a serum bilirubin>100 micromol/l (p = 0.001, OR ratio 3.70, 95% CI 1.84 to 7.47). Earlier referral (median serum bilirubin 78 micromol/l in the post-1998 era compared with 237 micromol/l in the pre-1998 era, p = 0.001) may be a contributory factor to improved survival. The strategies of combined en bloc reduced liver/small bowel transplantation and iLTx resulted in fewer deaths on the waiting list in the post-1998 era (2 deaths in post-1998 era v 12 deaths in pre-1998 era). The overall 3-year survival in the post-1998 era (69%) has improved compared with the pre-1998 era (31%; p<0.001) CONCLUSION: The changing characteristics at the time of referral, including earlier referral and innovative surgical strategies have resulted in improved long-term survival of children referred for ITx. PMID- 16705019 TI - Proteomics and genomics of microgravity. AB - Many serious adverse physiological changes occur during spaceflight. In the search for underlying mechanisms and possible new countermeasures, many experimental tools and methods have been developed to study microgravity caused physiological changes, ranging from in vitro bioreactor studies to spaceflight investigations. Recently, genomic and proteomic approaches have gained a lot of attention; after major scientific breakthroughs in the fields of genomics and proteomics, they are now widely accepted and used to understand biological processes. Understanding gene and/or protein expression is the key to unfolding the mechanisms behind microgravity-induced problems and, ultimately, finding effective countermeasures to spaceflight-induced alterations. Significant progress has been made in identifying the genes/proteins responsible for these changes. Although many of these genes and/or proteins were observed to be either upregulated or downregulated, however, no large-scale genomics and proteomics studies have been published so far. This review aims at summarizing the current status of microgravity-related genomics and proteomics studies and stimulating large-scale proteomics and genomics research activities. PMID- 16705020 TI - Quantitative PCR-based approach for rapid phage display analysis: a foundation for high throughput vascular proteomic profiling. AB - Functional proteomic strategies offer unique advantages over current molecular array approaches, as the epitopes identified can directly provide bioactive peptides for investigational and/or translational applications. The vascular endothelium is well suited to proteomic assessment by in vivo phage display, but extensive enrichment and sequencing steps limit its application for high throughput molecular profiling. To overcome these limitations we developed a quantitative PCR (Q-PCR) strategy to allow the rapid quantification of in vivo phage binding. Primers were designed for distinct clones selected from a defined phage pool to probe for age-associated changes in cardiac vascular epitopes. Sensitivity and specificity of the primer sets were tested and confirmed in vitro. Q-PCR quantification of phage in vivo confirmed the preferential homing of all phage clones to the young rather than old cardiac vasculature and demonstrated a close correlation with phage measurements previously determined using traditional bacterial-based titration methods. This Q-PCR approach provides quantification of phage within hours of phage injection and may therefore be used for rapid, high throughput analysis of binding of defined phage sequences both in vivo and in vitro, complementing nonbiased phage approaches for the proteomic mapping of vascular beds and other tissues. PMID- 16705021 TI - Fine mapping of Dyscalc1, the major genetic determinant of dystrophic cardiac calcification in mice. AB - Calcification of severely dystrophic muscle is occasionally observed in targeted mouse models of muscular dystrophy and cardiomyopathy. Intracellular calcium deposition occurs in necrotic myocytes in the absence of plasma calcium and phosphate imbalances. In the heart, this recessive trait is referred to as dystrophic cardiac calcinosis (DCC). We identified previously Dyscalc1, a major genetic determinant of DCC, in a 15.2-Mbp region on proximal chromosome 7. We report now further steps toward the identification of the Dyscalc1 gene by reverse genetics. Transferring the Dyscalc1 locus from susceptible mouse strain C3H/He onto a resistant C57BL/6 strain background, we generated congenic inbred strains B6.C3-(D7Mit56-D7Mit230) and B6.C3-(D7Nds5-D7Mit230). Three days after myocardial freeze-thaw injury, both strains exhibited calcification of necrotic lesions, confirming the pathogenetic relevance of Dyscalc1. Analysis of two (129S1 x C57BL/6) x 129S1 backcrosses allowed mapping of Dyscalc1 more precisely to a region spanning 0.76 Mbp between genes Fgf21 (39.70 Mbp) and Myod1 (40.46 Mbp). This interval contains 31 known and putative genes in three large, ancestral haplotypes shared by susceptible strains C3H/He, 129S1, and DBA/2. Thus we were able to exclude previously proposed candidate genes Bax and Hrc. Instead, a potential candidate may be the gene encoding the ATP-binding cassette C6. Mutations in the orthologous human ABCC6 gene cause pseudoxanthoma elasticum, or Gronblad-Strandberg syndrome, an elastic tissue disorder with cardiovascular calcifications. PMID- 16705022 TI - Cd14, Gbp1, and Pla2g2a: three major candidate genes for experimental IBD identified by combining QTL and microarray analyses. AB - Induction of inflammatory bowel (IBD)-like disease in mice by a targeted mutation in the Il10 gene (Il10(-/-)) is inbred strain dependent. C3H/HeJBir (C3) mice are colitis susceptible, whereas C57BL/6J (B6) mice are resistant. Genetic dissection of this susceptibility revealed 10 colitogenic quantitative trait loci (QTL). The aim of this study was to identify valuable candidate genes by a combination of QTL mapping and microarray analyses. Sixteen genes were differentially expressed between B6- and C3-Il10(-/-) mice and were located within the QTL intervals. Three major candidate genes (Cd14, Gbp1, Pla2g2a) showed prominent expression differences between B6- and C3-Il10(-/-) as well as between B6 and C3 wild-type mice, which was confirmed by semiquantitative or real-time RT-PCR. Because strain differences are known for Gbp1 and Pla2g2a, further analyses focused on Cd14. Western blot analysis revealed strain differences also on the protein level. Cd14 expression in animals with defective and intact Toll-like receptor (TLR)4 signaling (C3, C3H/HeN, B6, B6-Tlr4(tm1Aki)) make the TLR4 defect of C3 mice unlikely to be the reason for higher Cd14 expression. Less Cd14 expression in germ-free mice indicates a contribution of the microflora on Cd14 expression. Stimulation of naive peritoneal macrophages with bacterial antigens showed lower CD14 surface expression in B6 than in C3 mice. In conclusion, the large number of candidate genes was reduced to three major candidates that play an important role in inflammatory processes and immune response. Strain differences for them are already known or are shown in this study. PMID- 16705023 TI - Therapeutic approaches in autosomal dominant polycystic kidney disease (ADPKD): is there light at the end of the tunnel? PMID- 16705024 TI - Oral rifampin for prevention of S. aureus carriage-related infections in patients with renal failure--a meta-analysis of randomized controlled trials. AB - BACKGROUND: Rifampin has been studied as prophylaxis against Staphylococcus aureus-related infections in patients on dialysis. METHODS: We performed a meta analysis of randomized controlled trials (RCTs) that compared the effectiveness and safety of oral rifampin with another regimen or no therapy in reducing S. aureus-related infections in dialysis patients. RESULTS: Four RCTs evaluated oral rifampin (administered for 5 days every 3 months, or for 5 days once) as prophylaxis in dialysis patients. Oral rifampin with or without bacitracin was associated with less access-site infections with S. aureus compared with no treatment (odds ratio = 0.16, 95% confidence intervals: 0.06-0.44, 3 RCTs). There was no difference between prophylaxis with oral rifampin and topical mupirocin applied at the catheter site, for all studied outcomes, in the RCT comparing these regimens. Withdrawal from the study due to drug-related toxicity occurred in 7/107 (6.6%) of the studied patients with renal failure. Development of resistance of S. aureus to rifampin ranged from 0 to 18.2% (reported in three out of four included RCTs). CONCLUSION: Prophylactic use of oral rifampin reduces access-site infections with S. aureus in patients with renal failure undergoing dialysis. However, development of toxicity and antimicrobial resistance during the treatment with rifampin occur in considerable proportions of patients, limiting its use and supporting the guidelines that recommend the use of local antibiotics at the exit site, such as mupirocin, for these indications. The available data are rather limited and more studies should be performed to examine this important clinical question. PMID- 16705025 TI - The role of the alpha-1 adrenoceptor in modulating human mesangial cell matrix production. AB - BACKGROUND: The sympathetic nervous system is frequently activated in hypertension and may modify various aspects of renal function. Whether modulation of the sympathetic nervous system directly influences the development of renal fibrosis is yet to be established. The current study investigates the role of the alpha-1 adrenoceptor on human mesangial cell scarring. METHODS: Human mesangial cells were injured with macrophage-conditioned medium (MPCM) and treated with doxazosin for 1 or 3 days. RESULTS: alpha-1 Adrenoceptor antagonist doxazosin of 2 micromol/l reduced fibronectin protein in MPCM-injured female mesangial cells by 31 +/- 1.03% (P < 0.001) and by 9.5 +/- 0.3% (P = 0.01) in male mesangial cells. The differential response between sexes was significant (P = 0.004). alpha 1B Adrenoceptors were detected in human mesangial cells by reverse transcription polymerase chain reaction with expression in female cells being 87% higher than in males (P = 0.04). Injury with MPCM reduced alpha-1B adrenoceptor mRNA expression in both cell types. Doxazosin had no effect on the protein levels of transforming growth factor-beta (TGF-beta) or interleukin-1beta (IL-1beta), however, a small reduction in tumour necrosis factor-alpha (TNF-alpha) levels was observed. Doxazosin had no effect on the modulators of matrix turnover matrix metalloproteinases MMP3, MMP9 and tissue inhibitor of matrix metalloproteinases (TIMP-1), although a significant reduction in tissue plasminogen activator (tPA); (36.5 +/- 2.6%, P < 0.001) was observed. Doxazosin caused an up-regulation of kallikrein expression, both at mRNA and protein levels. Co-treatment with the bradykinin B2 receptor antagonist HOE140 was able to attenuate the effects of doxazosin treatment on fibronectin levels. CONCLUSION: These data suggest that inhibition of alpha-1B adrenoceptors in mesangial cells exerts an anti-fibrotic effect in a sex-specific manner via modulation of the kallikrein kinin/plasminogen activator system. PMID- 16705026 TI - Collapsing and non-collapsing focal segmental glomerulosclerosis in kidney transplants. AB - BACKGROUND: The aetiological and clinical associations of collapsing focal segmental glomerulosclerosis (cFSGS) following kidney transplantation (KTx) are poorly described. In this study, post-transplant cFSGS and non-collapsing FSGS (ncFSGS) were compared in recent KTx recipients. Evidence for intragraft viral infection was sought. METHODS: Twenty-nine cases of post-KTx FSGS were identified and classified as cFSGS (n = 10) or ncFSGS (n = 19). Biopsies were scored using Banff '97 criteria and subjected to in situ hybridization (ISH) for parvovirus B19 (pvB19), simian virus 40 (SV40) and BK virus (BKV). RESULTS: cFSGS and ncFSGS patients were comparable for age, gender, weight, delayed function, human leucocyte antigen (HLA) matching, acute rejection and median time to diagnosis. Deceased donor source was more common among cFSGS cases (70 vs 32%, P = 0.05). FSGS was recurrent in 2/10 cFSGS cases compared with 8/19 ncFSGS (P = NS). cFSGS was associated with more proteinuria (11.9 vs 7.2 g/day, P = 0.05) and higher serum creatinine (4.2 vs 1.9 mg/dl, P = 0.0001) at diagnosis. Plasmapheresis was used in two out of 10 cFSGS and seven out of 19 ncFSGS cases with treatment response in 0 of two and three of seven, respectively. Graft loss was more rapid with cFSGS compared with ncFSGS (P = 0.02). Histologically, cFSGS was associated with more severe chronic vascular abnormalities. All biopsies were negative for pvB19, SV40 and BKV by ISH. CONCLUSIONS: cFSGS following KTx presents with higher proteinuria, diminished renal function, more severe vascular disease and higher rate of graft loss compared with the non-collapsing form. There was no evidence for infection by pvB19 or polyomaviruses. PMID- 16705028 TI - How many "Me-Too" drugs are enough? The case of physician preferences for specific statins. AB - BACKGROUND: The increasing availability of "Me-Too" drugs has provided considerable treatment options for clinicians. However, the number of such drugs within a class that are actually used by clinicians has not been well studied. OBJECTIVE: To determine the number of different statins that individual physicians use in practice. METHODS: The Ontario Drug Benefit database was used to identify physicians who issued at least 10 incident statin prescriptions between October 2001 and May 2003 for patients aged 66 years and older. A preferred statin was defined for each physician, and the proportion of each physician's incident prescriptions written for that agent was determined. We then determined the number of different statins required to fill each physician's incident prescribing needs. RESULTS: A total of 3426 physicians wrote 73,571 incident statin prescriptions. The mean percentage of prescriptions written for each physician's preferred statin formulation was 73.7%. Repeat analysis to examine the proportion of prescriptions filled using each physician's top 2 statin formulations found that the average physician wrote the vast majority of his or her incident prescriptions (94.9%) for only 1 or 2 statins. Half of all physicians used, at most, 2 different statins for all incident prescribing, while 91.3% of physicians used, at most, 3 different statins for all of their incident prescribing. CONCLUSIONS: A high proportion of Ontario physicians issued the majority of their incident statin prescriptions for the same statin formulation. Most physicians required, at most, 3 different statins for all incident statin prescribing. PMID- 16705027 TI - Protective role of L-2-oxothiazolidine-4-carboxylic acid in cisplatin-induced renal injury. AB - BACKGROUND: Oxidative stress and inflammation are implicated in the pathogenesis of cisplatin-induced nephrotoxicity. l-2-oxothiazolidine-4-carboxylic acid (OTC) is a cysteine prodrug, and increases cellular glutathione (GSH). OTC is converted to cysteine by the intracellular enzyme, oxoprolinase. To date, the protective role of OTC on cisplatin-induced renal injury has not been investigated. The purpose of the present study was to examine the protective effect of OTC on cisplatin-induced renal injury and to examine the mechanism of its protection. METHODS: Mice were treated with cisplatin with or without administration of OTC. The generation of reactive oxygen species (ROS), expression of intercellular adhesion molecule (ICAM)-1 and monocyte chemoattractant protein (MCP)-1 were determined in the kidney using 2',7'-dichlorofluorescein diacetate, immunostaining or western blot analysis. Nuclear factor (NF)-kappaB activity, infiltration of F4/80-positive cells and apoptosis were also investigated in addition to renal function and histology using electrophoretic mobility shift assay, immunostaining, western blot analysis, uridine triphosphate (dUTP) nick end labelling or periodic acid-Schiff staining. The effect of OTC on superoxide dismutase activity and GSH level in cisplatin-treated normal adult human kidney (HK-2) cells were measured using assay kits. RESULTS: The administration of OTC resulted in a significant reduction of cisplatin-induced ROS production, the p65 subunit of NF-kappaB translocation into nucleus, expression of ICAM-1, caspase 3 activity, expression of MCP-1 and the infiltration of macrophages into renal tissue. OTC markedly ameliorated renal damage induced by cisplatin through antioxidant and anti-inflammatory effect. CONCLUSIONS: These results suggest that OTC can be a potential therapeutic agent in cisplatin-induced renal injury through decreasing the ROS levels and activation of NF-kappaB. PMID- 16705029 TI - Teduglutide for the treatment of short bowel syndrome. AB - OBJECTIVE: To summarize the pharmacology, development, and clinical application of teduglutide (ALX-0600), a glucagon-like peptide-2 (GLP-2) analog for the treatment of short bowel syndrome (SBS). DATA SOURCES: Clinical literature, including both primary sources and review articles, was accessed through a search of the MEDLINE databases (1980-March 2006). Key search terms included teduglutide, ALX-0600, glucagon-like peptide-2, short bowel syndrome, short gut, and intestinal adaptation. Clinical trial and drug data were supplied by the manufacturer, NPS Pharmaceuticals. STUDY SELECTION AND DATA EXTRACTION: Review articles, abstracts, and clinical studies related to GLP-2 and its analog, teduglutide, were analyzed. An evaluation of the research exploring teduglutide for the management of SBS was conducted. Relevant information was then selected. DATA SYNTHESIS: Research has revealed that administration of GLP-2 to patients following major small bowel resection improves intestinal adaptation and nutrient absorption. Teduglutide is an enzyme-resistant GLP-2 analog that shows promise in preventing intestinal injury, restoring mucosal integrity, and enhancing intestinal absorptive function. CONCLUSIONS: Data from ongoing clinical trials indicate that teduglutide may have the ability to enhance intestinal absorptive capacity in patients with SBS. Further studies and the completion of Phase III trials are necessary to determine the appropriate dosage and length of treatment for patients with SBS to gain optimal therapeutic benefit from this drug. PMID- 16705030 TI - Profile of Giorgio Parisi. PMID- 16705031 TI - Another weapon against amyloid. PMID- 16705032 TI - Understanding the bacterial polysaccharide antigenicity of Streptococcus agalactiae versus Streptococcus pneumoniae. AB - Bacterial surface capsular polysaccharides (CPS) that are similar in carbohydrate sequence may differ markedly in immunogenicity and antigenicity. The structural origin of these phenomena is poorly understood. Such a case is presented by the Gram-positive bacteria Streptococcus agalactiae (Group B Streptococcus; GBS) type III (GBSIII) and Streptococcus pneumoniae (Pn) type 14 (Pn14), which share closely related CPS sequences. Nevertheless, antibodies (Abs) against GBSIII rarely cross-react with the CPS from Pn14. To establish the origin for the variation in CPS antigenicity, models for the immune complexes of CPS fragments from GBSIII and Pn14, with the variable fragment (Fv) of a GBS-specific mAb (mAb 1B1), are presented. The complexes are generated through a combination of comparative Ab modeling and automated ligand docking, followed by explicitly solvated 10-ns molecular dynamics simulations. The relationship between carbohydrate sequence and antigenicity is further quantified through the computation of interaction energies using the Molecular Mechanics-Generalized Born Surface Area (MM-GBSA) method, augmented by conformational entropy estimates. Despite the electrostatic differences between Pn14 and GBSIII CPS, analysis indicates that entropic penalties are primarily responsible for the loss of affinity of the highly flexible Pn14 CPS for mAb 1B1. The similarity of the solution conformation of the relatively rigid GBSIII CPS with that in the immune complex characterizes the previously undescribed 3D structure of the conformational epitope. The analysis provides a comprehensive interpretation for a large body of biochemical and immunological data related to Ab recognition of bacterial polysaccharides and should be applicable to other Ab-carbohydrate interactions. PMID- 16705033 TI - Evolutionary tinkering with transposable elements. PMID- 16705034 TI - Rapid expansion of the spermatogonial stem cell tool box. PMID- 16705035 TI - Brain response to putative pheromones in lesbian women. AB - The progesterone derivative 4,16-androstadien-3-one (AND) and the estrogen-like steroid estra-1,3,5(10),16-tetraen-3-ol (EST) are candidate compounds for human pheromones. In previous positron emission tomography studies, we found that smelling AND and EST activated regions primarily incorporating the sexually dimorphic nuclei of the anterior hypothalamus, that this activation was differentiated with respect to sex and compound, and that homosexual men processed AND congruently with heterosexual women rather than heterosexual men. These observations indicate involvement of the anterior hypothalamus in physiological processes related to sexual orientation in humans. We expand the information on this issue in the present study by performing identical positron emission tomography experiments on 12 lesbian women. In contrast to heterosexual women, lesbian women processed AND stimuli by the olfactory networks and not the anterior hypothalamus. Furthermore, when smelling EST, they partly shared activation of the anterior hypothalamus with heterosexual men. These data support our previous results about differentiated processing of pheromone-like stimuli in humans and further strengthen the notion of a coupling between hypothalamic neuronal circuits and sexual preferences. PMID- 16705036 TI - The G12 family of heterotrimeric G proteins promotes breast cancer invasion and metastasis. AB - Although the prognosis for patients with early-stage breast cancer has improved, the therapeutic options for patients with locally advanced and metastatic disease are limited. To improve the treatment of these patients, the molecular mechanisms underlying breast cancer invasion and metastasis must be understood. In this study, we report that signaling through the G12 family of heterotrimeric G proteins (Galpha12 and Galpha13) promotes breast cancer cell invasion. Moreover, we demonstrate that inhibition of G12 signaling reduces the metastatic dissemination of breast cancer cells in vivo. Finally, we demonstrate that the expression of Galpha12 is significantly up-regulated in the earliest stages of breast cancer, implying that amplification of G12 signaling may be an early event in breast cancer progression. Taken together, these observations identify the G12 family proteins as important regulators of breast cancer invasion and suggest that these proteins may be targeted to limit invasion- and metastasis-induced patient morbidity and mortality. PMID- 16705038 TI - Lung adenocarcinomas induced in mice by mutant EGF receptors found in human lung cancers respond to a tyrosine kinase inhibitor or to down-regulation of the receptors. AB - Somatic mutations in exons encoding the tyrosine kinase domain of the epidermal growth factor receptor (EGFR) gene are found in human lung adenocarcinomas and are associated with sensitivity to the tyrosine kinase inhibitors gefitinib and erlotinib. Nearly 90% of the EGFR mutations are either short, in-frame deletions in exon 19 or point mutations that result in substitution of arginine for leucine at amino acid 858 (L858R). To study further the role of these mutations in the initiation and maintenance of lung cancer, we have developed transgenic mice that express an exon 19 deletion mutant (EGFR(DeltaL747-S752)) or the L858R mutant (EGFR(L858R)) in type II pneumocytes under the control of doxycycline. Expression of either EGFR mutant leads to the development of lung adenocarcinomas. Two weeks after induction with doxycycline, mice that express the EGFR(L858R) allele show diffuse lung cancer highly reminiscent of human bronchioloalveolar carcinoma and later develop interspersed multifocal adenocarcinomas. In contrast, mice expressing EGFR(DeltaL747-S752) develop multifocal tumors embedded in normal lung parenchyma with a longer latency. With mice carrying either EGFR allele, withdrawal of doxycycline (to reduce expression of the transgene) or treatment with erlotinib (to inhibit kinase activity) causes rapid tumor regression, as assessed by magnetic resonance imaging and histopathology, demonstrating that mutant EGFR is required for tumor maintenance. These models may be useful for developing improved therapies for patients with lung cancers bearing EGFR mutations. PMID- 16705037 TI - The Evf-2 noncoding RNA is transcribed from the Dlx-5/6 ultraconserved region and functions as a Dlx-2 transcriptional coactivator. AB - The identification of ultraconserved noncoding sequences in vertebrates has been associated with developmental regulators and DNA-binding proteins. One of the first of these was identified in the intergenic region between the Dlx-5 and Dlx 6 genes, members of the Dlx/dll homeodomain-containing protein family. In previous experiments, we showed that Sonic hedgehog treatment of forebrain neural explants results in the activation of Dlx-2 and the novel noncoding RNA (ncRNA), Evf-1. In this report, we show that the Dlx-5/6 ultraconserved region is transcribed to generate an alternatively spliced form of Evf-1, the ncRNA Evf-2. Evf-2 specifically cooperates with Dlx-2 to increase the transcriptional activity of the Dlx-5/6 enhancer in a target and homeodomain-specific manner. A stable complex containing the Evf-2 ncRNA and the Dlx-2 protein forms in vivo, suggesting that the Evf-2 ncRNA activates transcriptional activity by directly influencing Dlx-2 activity. These experiments identify a novel mechanism whereby transcription is controlled by the cooperative actions of an ncRNA and a homeodomain protein. The possibility that a subset of vertebrate ultraconserved regions may function at both the DNA and RNA level to control key developmental regulators may explain why ultraconserved sequences exhibit 90% or more conservation even after 450 million years of vertebrate evolution. PMID- 16705041 TI - Effects of 4-n-nonylphenol and tamoxifen on salmon gonadotropin-releasing hormone, estrogen receptor, and vitellogenin gene expression in juvenile rainbow trout. AB - Alkylphenols such as nonylphenol (NP) are one of a wide variety of environmental chemicals reported to have estrogenic effects in both in vitro and in vivo studies. Induction of hepatic vitellogenin (Vg) gene expression is widely used as a biomarker for xenoestrogen exposure in fish. However, little work has been done to characterize the molecular effects of xenoestrogens on other potential target organs such as the brain. To evaluate brain and liver effects of 4-n-nonylphenol (4-NP), juvenile rainbow trout (Oncorhynchus mykiss) were exposed to waterborne 4 NP or 17beta-estradiol (E(2)). Changes in mRNA levels of salmon gonadotropin releasing hormone (sGnRH) and estrogen receptor alpha (ERalpha) isoforms in the brain and ERalpha isoforms and Vg in the liver were measured after 3 and 6 days of exposure, with the help of a relative RT-PCR-based quantification method. Fish were treated with increasing doses of 4-NP ranging from 0.01 to 10 microM (2.2 microg/l to 2.2 mg/l), and results were compared to the effect of E(2) or tamoxifen, a specific ER modulator. In liver, E(2) and the highest doses of 4-NP increased Vg and ERalpha long-isoform mRNA levels within 3 or 6 days of exposure, but 4-NP did not have any effect on ERalpha short-isoform transcription level. In the brain, 4-NP reduced sGnRH2 gene expression in a dose-dependent manner, but did not modify sGnRH1 or ERalpha mRNA levels. PMID- 16705039 TI - The locus control region is required for association of the murine beta-globin locus with engaged transcription factories during erythroid maturation. AB - We have examined the relationship between nuclear localization and transcriptional activity of the endogenous murine beta-globin locus during erythroid differentiation. Murine fetal liver cells were separated into distinct erythroid maturation stages by fluorescence-activated cell sorting, and the nuclear position of the locus was determined at each stage. We find that the beta globin locus progressively moves away from the nuclear periphery with increasing maturation. Contrary to the prevailing notion that the nuclear periphery is a repressive compartment in mammalian cells, beta(major)-globin expression begins at the nuclear periphery prior to relocalization. However, relocation of the locus to the nuclear interior with maturation is accompanied by an increase in beta(major)-globin transcription. The distribution of nuclear polymerase II (Pol II) foci also changes with erythroid differentiation: Transcription factories decrease in number and contract toward the nuclear interior. Moreover, both efficient relocalization of the beta-globin locus from the periphery and its association with hyperphosphorylated Pol II transcription factories require the locus control region (LCR). These results suggest that the LCR-dependent association of the beta-globin locus with transcriptionally engaged Pol II foci provides the driving force for relocalization of the locus toward the nuclear interior during erythroid maturation. PMID- 16705040 TI - p27kip1 independently promotes neuronal differentiation and migration in the cerebral cortex. AB - The generation of neurons by progenitor cells involves the tight coordination of multiple cellular activities, including cell cycle exit, initiation of neuronal differentiation, and cell migration. The mechanisms that integrate these different events into a coherent developmental program are not well understood. Here we show that the cyclin-dependent kinase inhibitor p27(Kip1) plays an important role in neurogenesis in the mouse cerebral cortex by promoting the differentiation and radial migration of cortical projection neurons. Importantly, these two functions of p27(Kip1) involve distinct activities, which are independent of its role in cell cycle regulation. p27(Kip1) promotes neuronal differentiation by stabilizing Neurogenin2 protein, an activity carried by the N terminal half of the protein. p27(Kip1) promotes neuronal migration by blocking RhoA signaling, an activity that resides in its C-terminal half. Thus, p27(Kip1) plays a key role in cortical development, acting as a modular protein that independently regulates and couples multiple cellular pathways contributing to neurogenesis. PMID- 16705042 TI - A manganese-enhanced diet alters brain metals and transporters in the developing rat. AB - Manganese (Mn) neurotoxicity in adults can result in psychological and neurological disturbances similar to Parkinson's disease, including extrapyramidal motor system defects and altered behaviors. However, virtually nothing is known regarding excess Mn accumulation during central nervous system development. Developing rats were exposed to a diet high in Mn via maternal milk during lactation (PN4-21). The high Mn diet resulted in changes in hematological parameters similar to those seen with iron (Fe) deficiency in dams (decreased plasma Fe; increased plasma transferrin [Tf]) and pups (decreased hemoglobin [Hb] and plasma Fe; increased plasma Tf and total iron binding capacity). Mn-exposed pups showed an increase in brain Mn, chromium, and zinc concurrent with a decrease in brain Fe. In conjunction with the altered transport and distribution of essential metals within the brain, there was enhanced protein expression of the divalent metal transporter-1 (DMT-1) and transferrin receptor (TfR) overall in the brain; there was a general increase in each region analyzed (cerebellum, cortex, hippocampus, midbrain, and striatum). Neurochemical changes were observed as an increase in gamma-aminobutyric acid (GABA) and the ratio of GABA to glutamate, indicating enhanced inhibitory transmission in the brain. The results of this study demonstrate that developing rats undergo alterations in the transport and distribution of essential metals translating to neurochemical perturbations after maternal exposure to a diet supplemented with excess levels of Mn. PMID- 16705043 TI - Low-dose infliximab treatment for ankylosing spondylitis--clinically- and cost effective. AB - OBJECTIVES: Infliximab has been shown to be effective in the treatment of ankylosing spondylitis (AS) when treated in a dose of 5 mg/kg at 6 weekly intervals. This dose of infliximab has not been determined by any structured randomized trials and has significant cost implications. We describe our experience of treating AS with low-dose infliximab (3mg/kg at 8 weekly intervals). The efficacy and cost implications are discussed. METHODS: Patients who had active AS [Bath AS Disease Activity Index (BASDAI) > or = 4] were treated with infliximab 3 mg/kg at 0, 2, 6 weeks and thereafter at 8 weekly intervals. Response to treatment was defined as 50% improvement in BASDAI. Other response criteria such as ASAS 20, 40 and five of the six criteria were also assessed. Direct drug costs for infliximab were determined. RESULTS: Twenty-two consecutive AS patients received infliximab. All 22 completed treatment for 3 months, 15 patients for 6 months and 14 for 12 months. Mean age was 45 years (range 21-62) and mean disease duration 14.5 years (range 2-43). Of the patients, 54% achieved a 50% BASDAI response at 3 months and the benefit was sustained at 12 months in 63%. Similar response rate was seen with the other assessment criteria. Direct drug costs were significantly lower when low-dose infliximab regimen was used. CONCLUSIONS: Low-dose infliximab (3 mg/kg at 8 weekly infusions) is effective in the treatment of AS. Higher doses are required in a small proportion of patients when treatment is only partially effective. Titrating the dose and frequency of infusions may be required in individual patients to achieve optimal response. Using low-dose infliximab has significant economic implications. PMID- 16705044 TI - Pharmacoeconomic considerations in the treatment of psoriatic arthritis. PMID- 16705045 TI - Wegener's granulomatosis presenting with an acute ST-elevation myocardial infarct (STEMI). PMID- 16705047 TI - Macrophage migration inhibitory factor and glucocorticoid sensitivity. AB - Glucocorticoids (GCs) are widely used in the treatment of inflammatory diseases including rheumatoid arthritis (RA). Treatment with GC is associated with significant dose-dependent side-effects. The pro-inflammatory cytokine macrophage migration inhibitory factor (MIF) has emerged in recent years as a candidate factor which could regulate GC sensitivity. MIF is induced by GC, and is able to override anti-inflammatory actions of GCs. In this review, we summarize the pro inflammatory actions of MIF with respect to RA, describe the interactions between MIF and GC and examine new evidence, which identifies MIF as a specific target for steroid sparing. PMID- 16705046 TI - Predictors of response to anti-TNF-alpha therapy among patients with rheumatoid arthritis: results from the British Society for Rheumatology Biologics Register. AB - BACKGROUND: Anti-tumour necrosis factor-alpha (TNF-alpha) therapies represent an important advancement in therapy for rheumatoid arthritis (RA). However, there remains a proportion of patients who do not improve despite therapy. These drugs are expensive and have the potential of serious toxicity. Therefore, it would be ideal to predict the patients who will respond, so that the use of these drugs can be targetted. OBJECTIVE: To identify the clinical factors present at the start of anti-TNF-alpha therapy that are associated with response at 6 months in patients with RA. METHODS: The British Society for Rheumatology (BSR) Biologics Register collects detailed data on all patients with a rheumatic disease receiving biologic therapy in the UK. We studied all patients with RA who had started etanercept (ETA) or infliximab (INF) and had achieved a minimum 6 months follow-up by 1 October, 2004. The disease status at the baseline and at 6 months was assessed using the Disease Activity Score (DAS28). The response was classified according to the European League against Rheumatism (EULAR) improvement criteria. The effect of baseline characteristics on response was studied using multivariate ordinal logistic regression. RESULTS: 2879 patients were included in this analysis (1267 ETA, 1612 INF). At the start of therapy, the mean age was 55 yrs, disease duration 14 yrs, baseline DAS28 6.7 and health assessment questionnaire (HAQ) 2.1. In all, 28% of ETA and 86% of INF patients were receiving methotrexate. After 6 months, 18% had a good EULAR response, of whom 9% were considered to be in remission and 50% had a moderate response. There was no overall difference in response rate between the two anti-TNF-alpha therapies. A higher baseline HAQ score correlated with a lower response rate while a better response was associated with the current use of NSAIDs and the use of methotrexate (MTX), although the latter only reached statistical significance with ETA [OR 1.82 (95% CI 1.38-2.40)]. There was a lower response rate among current smokers, particularly in patients receiving INF [OR 0.77 (95% CI 0.60 0.99)]. Age, disease duration, rheumatoid factor and the previous number of disease-modifying antirheumatic drugs (DMARDs) did not predict response to either drug. However, females were less likely to achieve remission. CONCLUSIONS: These data support an improved outcome among patients receiving MTX in combination with anti-TNF-alpha therapies. However, the most disabled patients were less likely to respond, despite concurrent MTX. The benefits of NSAIDs may reflect the relative absence of comorbidities in patients who can tolerate these drugs or the continuing presence of reversible inflammatory symptoms. The association of smoking and poor outcome with INF is a novel finding and may reflect alterations in pharmacokinetics. The inability of other baseline disease characteristics to predict the outcome suggests that other factors, including potential genetic differences in drug metabolism, may be influencing the response to anti-TNF-alpha therapies. PMID- 16705048 TI - Isoprostanes as a tool to investigate oxidative stress in scleroderma spectrum disorders--advantages and limitations. PMID- 16705050 TI - Overlapping humoral autoimmunity links rheumatic fever and the antiphospholipid syndrome. AB - OBJECTIVE: Rheumatic fever (RF) and the antiphospholipid syndrome (APS) are autoimmune diseases that share similar cardiac and neurological pathologies. We assessed the presence of shared epitopes between M protein, N-acetyl-beta-D glucosamine (GlcNAc) and beta2 glycoprotein-I (beta2GPI), the pathogenic molecules engaged in these autoimmune conditions. METHODS: Sera from the APS patients were affinity-purified on beta2GPI and beta2GPI-related peptide columns. Sera from RF patients were affinity-purified on protein G column. The beta2GPI and M protein-related peptides were prepared by conventional solid-phase peptide synthesis. The enzyme-linked immunosorbent assay direct binding and inhibition studies were performed on the RF and APS sera for the presence, and cross reactivity, of antibodies against beta2GPI, beta2GPI-related peptides, streptococcal M protein, M-derived peptides and GlcNAc. RESULTS: Antibodies (Abs) to beta2GPI were found in 24.4% of 90 RF patients. Antibodies against various beta2GPI-related peptides were found in 1.1-36.7% of the patients. The immunoglobulin G sera from RF patients possessed significant anti-beta2GPI activity, while sera from APS patients contained a considerable anti streptococcal M protein as well as anti-GlcNAc activity. Furthermore, affinity purified anti-beta2GPI and anti-beta2GPI-related peptide Abs from APS patients cross-reacted with streptococcal M protein and M5 peptide, while beta2GPI and beta2GPI-related peptides inhibited anti-streptococcal M protein activity from RF patients. The results were confirmed by immunoblot analyses. The beta2GPI also inhibited anti-GlcNAc activity from APS patients with chorea. CONCLUSIONS: The results of our study, showing a considerable overlap of humoral immunity in RF and APS, support a hypothesis that common pathogenic mechanisms underlie the development of cardiac valve lesions and Central Nervous System abnormalities in both diseases. PMID- 16705051 TI - Digital thermal hyperaemia impairment does not relate to skin fibrosis or macrovascular disease in systemic sclerosis. AB - OBJECTIVES: Thermal hyperaemia is impaired in patients with systemic sclerosis (SSc). The objective of these studies was to determine whether this was consecutive to skin fibrosis, microangiopathy or macroangiopathy. METHODS: Using laser Doppler flowmetry, we first compared the thermal hyperaemia on the third left finger pad and on the left forearm in 21 patients with non-diffuse systemic sclerosis (SSc), in comparison with primary Raynaud's phenomenon and healthy volunteers. Second, we tested whether the altered thermal hyperaemia correlated to the digital pressure index at baseline, and following the thermal challenge. RESULTS: In the first study, thermal hyperaemia of the finger pad was impaired in terms of both amplitude and kinetics, but not on the forearm in patients with SSc. In the seven SSc patients without cutaneous fibrosis, the response was similarly altered in terms of amplitude and kinetics. In the second study, we observed a weak correlation between the digital systolic blood pressure index. However, in the 15 SSc patients tested at 44 degrees C, the median digital systolic blood pressure index was 1.04 (0.84-1.24) at baseline vs 1.08 (0.87 1.29) at 44 degrees C (NS), while seven of them had an abnormal response in terms of kinetic. Furthermore, only one patient showed a clear-cut decrease in digital systolic blood pressure at 44 degrees C. CONCLUSION: In patients with SSc, digital thermal hyperaemia is impaired, but does not relate to the skin fibrosis or to an associated macroangiopathy in most cases. Further studies are required to determine whether its impairment reflects a functional or structural microvascular damage. PMID- 16705052 TI - Meeting the needs of increasing numbers of medical students--a best practise approach. PMID- 16705054 TI - Effect of insulin on human skeletal muscle protein synthesis is modulated by insulin-induced changes in muscle blood flow and amino acid availability. AB - Insulin promotes muscle anabolism, but it is still unclear whether it stimulates muscle protein synthesis in humans. We hypothesized that insulin can increase muscle protein synthesis only if it increases muscle amino acid availability. We measured muscle protein and amino acid metabolism using stable-isotope methodologies in 19 young healthy subjects at baseline and during insulin infusion in one leg at low (LD, 0.05), intermediate (ID, 0.15), or high (HD, 0.30 mUxmin(-1)x100 ml(-1)) doses. Insulin was infused locally to induce muscle hyperinsulinemia within the physiological range while minimizing the systemic effects. Protein and amino acid kinetics across the leg were assessed using stable isotopes and muscle biopsies. The LD did not affect phenylalanine delivery to the muscle (-9 +/- 18% change over baseline), muscle protein synthesis (16 +/- 26%), breakdown, or net balance. The ID increased (P < 0.05) phenylalanine delivery (+63 +/- 38%), muscle protein synthesis (+157 +/- 54%), and net protein balance, with no change in breakdown. The HD did not change phenylalanine delivery (+12 +/- 11%) or muscle protein synthesis (+9 +/- 19%), and reduced muscle protein breakdown (-17 +/- 15%), thus improving net muscle protein balance but to a lesser degree than the ID. Changes in muscle protein synthesis were strongly associated with changes in muscle blood flow and phenylalanine delivery and availability. In conclusion, physiological hyperinsulinemia promotes muscle protein synthesis as long as it concomitantly increases muscle blood flow, amino acid delivery and availability. PMID- 16705055 TI - Angiotensin II induces monocyte chemoattractant protein-1 expression via a nuclear factor-kappaB-dependent pathway in rat preadipocytes. AB - Both monocyte chemoattractant protein-1 (MCP-1), a member of chemokine family, and angiotensinogen, a precursor of angiotensin (ANG) II, are produced by adipose tissue and increased in obese state. MCP-1 has been shown to decrease insulin stimulated glucose uptake and several adipogenic genes expression in adipocytes in vitro, suggesting its pathophysiological significance in obesity. However, the pathophysiological interaction between MCP-1 and ANG II in adipose tissue remains unknown. The present study was undertaken to investigate the potential mechanisms by which ANG II affects MCP-1 gene expression in rat primary cultured preadipocytes and adipose tissue in vivo. ANG II significantly increased steady state MCP-1 mRNA levels in a time- and dose-dependent manner. The ANG II-induced MCP-1 mRNA and protein expression was completely abolished by ANG II type 1 (AT1) receptor antagonist (valsartan). An antioxidant/NF-kappaB inhibitor (pyrrolidine dithiocarbamate) and an inhibitor of 1kappaB-alpha phosphorylation (Bay 11-7085) also blocked ANG II-induced MCP-1 mRNA expression. ANG II induced translocation of NF-kappaB p65 subunit from cytoplasm to nucleus by immunocytochemical study. Luciferase assay using reporter constructs containing MCP-1 promoter region revealed that two NF-kappaB binding sites in its enhancer region were essential for the ANG II-induced promoter activities. Furthermore, basal mRNA and protein of MCP-1 during preadipocyte differentiation were significantly greater in preadipocytes than in differentiated adipocytes, whose effect was more pronounced in the presence of ANG II. Exogenous administration of ANG II to rats led to increased MCP-1 expression in epididymal, subcutaneous, and mesenteric adipose tissue. In conclusion, our present study demonstrates that ANG II increases MCP-1 gene expression via ANG II type 1 receptor-mediated and NF-kappaB-dependent pathway in rat preadipocytes as well as adipose MCP-1 expression in vivo. Thus the augmented MCP-1 expression by ANG II in preadipocytes may provide a new link between obesity and cardiovascular disease. PMID- 16705056 TI - Inhibition of calpain results in impaired contraction-stimulated GLUT4 translocation in skeletal muscle. AB - It was previously found that transgenic mice that overexpress the calpain inhibitor calpastatin (CsTg) have an approximately 3-fold increase in GLUT4 protein in their skeletal muscles. Despite the increase in GLUT4, which appears to be due to inhibition of its proteolysis by calpain, insulin-stimulated glucose transport is not increased in CsTg muscles. PKB (Akt) protein level is reduced approximately 60% in CsTg muscles, suggesting a possible mechanism for the relative insulin resistance. Muscle contractions stimulate glucose transport by a mechanism that is independent of insulin signaling. The purpose of this study was to test the hypothesis that the threefold increase in GLUT4 in CsTg would result in a large increase in contraction-stimulated glucose transport. CAMKII and AMPK mediate steps in the contraction-stimulated pathway. The protein levels of AMPK and CAMKII were increased three- to fourfold in CsTg muscles, suggesting that these proteins are also calpain substrates. Despite the large increases in GLUT4, AMPK, and CAMKII, contraction-stimulated GLUT4 translocation and glucose transport were not increased above wild-type values. These findings suggest that inhibition of calpain results in impairment of a step in the GLUT4 translocation process downstream of the insulin- and contraction-signaling pathways. They also provide evidence that CAMKII and AMPK are calpain substrates. PMID- 16705057 TI - Lipid metabolism and secretory function of porcine intramuscular adipocytes compared with subcutaneous and perirenal adipocytes. AB - The function of adipocytes interspersed between myofiber fasciculi in skeletal muscle physiology and physiopathology is poorly documented. Because regional differences in adipocyte features have been reported in various species, we hypothesized that lipid metabolism and secretory function of intramuscular (IM) adipocytes differ from that of nonmuscular adipocytes. In the present study, adipocytes isolated from trapezius muscle were compared with subcutaneous and perirenal adipocytes in growing pigs. Between 80 and 210 days of age, gene expressions and/or activities of enzymes involved in lipogenesis or lipolysis were much lower (P < 0.05) in adipocytes isolated from muscle than in those from other locations. Insulin-induced lipogenesis and lipolytic efficiency after catecholamine addition were also the lowest (P < 0.05) in IM adipocytes. In these cells, the age-related increase (+300%) in the ratio of mRNA levels of fatty acid synthase to hormone-sensitive lipase paralleled the enlargement of adipocyte diameters (+70%, P < 0.05) and the increase in lipid content in muscle (+135%, P < 0.05) during growth. Expressions of genes coding for leptin, adiponectin, and IGF-I, as well as for various hormonal receptors, were lower (P < 0.05) in IM adipocytes than in other adipocytes, whereas levels of TNF-alpha mRNA did not differ between sites. Interestingly, IGF-II mRNA levels were higher (P < 0.05) in IM adipocytes than in other adipocytes. These data support the view that IM fat is not just an ectopic extension of other fat locations but displays specific biological features during growth. PMID- 16705058 TI - Parenteral and enteral metabolism of anaplerotic triheptanoin in normal rats. AB - A new chronic treatment for inherited disorders of long-chain fatty acid oxidation involves administering up to one-third of dietary calories as triheptanoin, a medium-odd-chain triglyceride (Roe CR, Sweetman L, Roe DS, David F, and Brunengraber H. J Clin Invest 110: 259-269, 2002). Heptanoate and C(5) ketone bodies derived from its partial oxidation in liver are precursors of anaplerotic propionyl-CoA in peripheral tissues. It was hypothesized that increasing anaplerosis in peripheral tissues would boost energy production. In the present study, we tested the potential of a triheptanoin emulsion as an intravenous nutrient. Normal rats were infused with triheptanoin intravenously or intraduodenally at up to 40% of caloric requirement. The blood concentration ratio (heptanoate/C(5)-ketone bodies) was high with intravenous and low with intraduodenal triheptanoin infusion. During intravenous infusion of triheptanoin, lipolysis was stimulated but appeared compensated by fatty acid reesterification. During intraduodenal infusion of triheptanoin, lipolysis was not stimulated. Our data support the hypothesis that intravenous triheptanoin could be used to treat decompensated patients with long-chain fatty acid oxidation disorders. PMID- 16705059 TI - Relationship between insulin sensitivity and in vivo mitochondrial function in skeletal muscle. AB - Recent data have shown that individuals with low insulin sensitivity (S(I)) also have reduced whole body maximal oxygen uptake. The objectives of this study were to determine 1) whether muscle mitochondrial function was independently related to S(I) after being adjusted for known determinants of S(I) and 2) whether lower S(I) among African-American (AA) vs. Caucasian-American (CA) women was due to lower muscle mitochondrial function among AA women. Subjects were 37 CA and 22 AA premenopausal women (age: 33.6 +/- 6.3 yr). Mitochondrial function [time constant of ADP (ADP(tc))] was assessed during a 90-s unilateral isometric contraction using (31)P magnetic resonance spectroscopy, S(I) with an intravenous glucose tolerance test, body composition by dual-energy X-ray absorptiometry, and visceral adipose tissue (VAT) with computed tomography. ANOVA was used to compare AA and CA groups, and multiple linear regression modeling was used to identify independent predictors of S(I). Between-race comparisons indicated that muscle oxidative capacity was lower among AAs vs. CAs (ADP(tc): 25.6 +/- 9.8 vs. 21.4 +/ 9.9 s). Multiple linear regression models for the dependent variable S(I) contained 1) VAT and race and 2) VAT, race, and ADP(tc). Significant independent effects for all predictor variables were observed in both the first (r(2) = 0.345) and second (r(2) = 0.410) models. The partial correlation for race was lower in the second model (-0.404 vs. -0.300), suggesting that muscle mitochondrial function contributed to the racial difference in S(I). Lower muscle mitochondrial function among AAs may in part explain lower S(I) among them. PMID- 16705060 TI - The adipose tissue triglyceride lipase ATGL/PNPLA2 is downregulated by insulin and TNF-alpha in 3T3-L1 adipocytes and is a target for transactivation by PPARgamma. AB - The minimal adipose phenotype of hormone-sensitive lipase (HSL)-null mice suggested that other hormonally responsive lipase(s) were present in adipocytes. Recent studies have characterized a new adipose tissue triglyceride lipase, ATGL/PNPLA2/destnutrin/iPLA2zeta/TTS2.2 (ATGL). We had previously cloned a novel adipose-enriched transcript by differential screening and recently determined its identity with murine ATGL. We report here on the regulation of ATGL by TNF-alpha and insulin in 3T3-L1 adipocytes and identify ATGL as a target for transcriptional activation by the key adipogenic transcription factor PPARgamma. Insulin at 100 nM resulted in a marked decrease in ATGL transcript that was effectively blocked by inhibitors for PI 3-kinase and p70 ribosomal protein S6 kinase. TNF-alpha treatment decreased ATGL transcript in a time-dependent manner that paralleled TNF-alpha downregulation of PPARgamma with a maximal decrease noted by 6 h. TNF-alpha effects on ATGL were attenuated by pretreatment with PD 98059, LY-294002, or rapamycin, suggesting involvement of the p44/42 MAP kinase, PI 3-kinase, and p70 ribosomal protein S6 kinase signals. To study transcriptional regulation of ATGL, we cloned 2,979 bp of the murine ATGL 5' flanking region. Compared with promoterless pGL2-Basic, the -2979/+21 ATGL luciferase construct demonstrated 120- and 40-fold increases in activity in white and brown adipocytes, respectively. Luciferase reporter activities for a series of eight ATGL promoter deletions revealed that the -928/+21, -1738/+21, 1979/+21, and -2979/+21 constructs were transactivated by PPARgamma. Our findings identify the novel lipase ATGL to be a target gene for TNF-alpha and insulin action in adipocytes and reveal that it is subject to transcriptional control by PPARgamma-mediated signals. PMID- 16705061 TI - Overexpression of acyl-CoA synthetase-1 increases lipid deposition in hepatic (HepG2) cells and rodent liver in vivo. AB - Accumulation of intracellular lipid in obesity is associated with metabolic disease in many tissues including liver. Storage of fatty acid as triglyceride (TG) requires the activation of fatty acids to long-chain acyl-CoAs (LC-CoA) by the enzyme acyl-CoA synthetase (ACSL). There are five known isoforms of ACSL (ACSL1, -3, -4, -5, -6), which vary in their tissue specificity and affinity for fatty acid substrates. To investigate the role of ACSL1 in the regulation of lipid metabolism, we used adenoviral-mediated gene transfer to overexpress ACSL1 in the human hepatoma cell-line HepG2 and in liver of rodents. Infection of HepG2 cells with the adenoviral construct AdACSL1 increased ACSL activity >10-fold compared with controls after 24 h. HepG2 cells overexpressing ACSL1 had a 40% higher triglyceride (TG) content (93 +/- 3 vs. 67 +/- 2 nmol/mg protein in controls, P < 0.05) after 24-h exposure to 1 mM oleate. Furthermore, ACSL1 overexpression produced a 60% increase in cellular LCA-CoA content (160 +/- 6 vs. 100 +/- 6 nmol/g protein in controls, P < 0.05) and increased [(14)C]oleate incorporation into TG without significantly altering fatty acid oxidation. In mice, AdACSL1 administration increased ACSL1 mRNA and protein more than fivefold over controls at 4 days postinfection. ACSL1 overexpression caused a twofold increase in TG content in mouse liver (39 +/- 4 vs. 20 +/- 2 mumol/g wet wt in controls, P < 0.05), and overexpression in rat liver increased [1-(14)C]palmitate clearance into liver TG. These in vitro and in vivo results suggest a pivotal role for ACSL1 in regulating TG synthesis in liver. PMID- 16705063 TI - Deficiency of carbohydrate-activated transcription factor ChREBP prevents obesity and improves plasma glucose control in leptin-deficient (ob/ob) mice. AB - The transcription factor carbohydrate response element-binding protein (ChREBP) mediates insulin-independent, glucose-stimulated gene expression of multiple liver enzymes responsible for converting excess carbohydrate to fatty acids for long-term storage. To investigate ChREBP's role in the development of obesity and obesity-associated metabolic dysregulation, ChREBP-deficient mice were intercrossed with ob/ob mice. As a result of deficient leptin expression, ob/ob mice overeat, become obese and resistant to insulin, and display marked elevations in hepatic lipogenesis, gluconeogenesis, and plasma glucose and triglycerides. mRNA expression of all hepatic lipogenic enzymes was significantly lower in ob/ob-ChREBP-/- than in ob/ob mice, resulting in decreased hepatic fatty acid synthesis and normalization of plasma free fatty acid and triglyceride levels. Overall weight gain in addition to adiposity was reduced in the doubly deficient mice. The former was largely attributable to decreased food intake and may result from decreased hypothalamic expression of the appetite-stimulating neuropeptide agouti-related protein. mRNA expression and activity of gluconeogenic enzymes also was lower in the doubly deficient mice, contributing to significantly lower blood glucose levels. The results of this study suggest that inactivation of ChREBP expression not only reduces fat synthesis and obesity in ob/ob mice but also results in improved glucose tolerance and appetite control. PMID- 16705062 TI - Differential coupling of beta3A- and beta3B-adrenergic receptors to endogenous and chimeric Galphas and Galphai. AB - Chimeric G proteins made by replacing the COOH-terminal heptapeptide of G(alpha)q with the COOH-terminal heptapeptide of G(alpha)s or G(alpha)i were used to assess the relative coupling of beta(3)-adrenergic receptor (beta(3)-AR) splice variants (beta(3A) and beta(3B)) to G(alpha)s and G(alpha)i. The G(alpha)q/s and G(alpha)q/i chimeras transformed the response to receptor activation from regulation of adenylyl cyclase to mobilization of intracellular calcium (Ca(2+)(i)). Complementary high-throughput and single-cell approaches were used to evaluate agonist-induced coupling of the receptor to the G protein chimeras. In cells stably transformed with rat beta(3)-AR, transfected with the G protein chimeras, and evaluated using a scanning fluorometer, beta(3)-AR-induced coupling to G(alpha)q/s produced a rapid eightfold increase in Ca(2+)(i) followed by a slow decay to levels 25% above baseline. G(alpha)q/i also linked rat beta(3)-AR to mobilization of Ca(2+)(i) in a similar time- and agonist-dependent manner, but the net 2.5-fold increase in Ca(2+)(i) was only 30% of the response obtained with G(alpha)q/s. Activation of the rat beta(3)-AR also increased GTP binding to endogenous G(alpha)i threefold in membranes from CHO cells stably transformed with the receptor. A complementary single-cell imaging approach was used to assess the relative coupling of mouse beta(3A)- and beta(3B)-AR to G(alpha)i under conditions established to produce equivalent agonist-dependent coupling of the receptor splice variants to G(alpha)q/s and to increases in intracellular cAMP through endogenous G(alpha)s. The beta(3A)- and beta(3B)-AR coupled equivalently to G(alpha)q/i, but the temporal patterns of Ca(2+)(i) mobilization indicated that coupling was significantly less efficient than coupling to G(alpha)q/s. Collectively, these findings indicate less efficient but equivalent coupling of beta(3A)- and beta(3B)-AR to G(alpha)i vs. G(alpha)s and suggest that differential expression of the splice variants would not produce local differences in signaling networks linked to beta(3)-AR activation. PMID- 16705064 TI - Insulin sensitivity is preserved despite disrupted endothelial function. AB - It is well established that endothelial dysfunction and insulin resistance go hand in hand. However, it is unclear whether endothelial dysfunction per se is sufficient to impair insulin-mediated glucose uptake. We have previously reported that 4 wk of administration of the human immunodeficiency virus (HIV)-1 protease inhibitor indinavir to HIV-negative subjects induces endothelial dysfunction. Hence, we hypothesized that indinavir-induced endothelial dysfunction was associated with impaired insulin-mediated glucose disposal. We measured insulin mediated glucose disposal at the level of the whole body, skeletal muscle, and vasculature by performing hyperinsulinemic euglycemic clamp, and vascular function studies, in a separate group of HIV-negative healthy nonobese subjects (n = 13) before and after 4 wk of daily oral indinavir. Four weeks of indinavir resulted in a 113 +/- 29% (P < 0.01) reduction of endothelium-dependent vasodilation, consistent with our earlier findings. In addition, there was a significant impairment of insulin-mediated vasodilation (101 +/- 14% before indinavir vs. 35 +/- 15% after indinavir; P < 0.05). However, there was no significant change in insulin-mediated glucose disposal at the level of the whole body (8.9 +/- 0.5 before indinavir vs. 8.5 +/- 0.6 mgxkg(-1)xmin(-1) after indinavir; P = 0.4), or skeletal muscle. Furthermore, in a separate group of four HIV-negative healthy nonobese subjects, we found that 4 wk of indinavir has no sustained effect on insulin-stimulated glucose uptake in adipose tissue. Thus our findings indicate that 1) endothelial dysfunction alone is insufficient to impair insulin-mediated glucose disposal, and 2) indinavir-induced endothelial dysfunction is likely due to a direct effect of the drug on the endothelium and is not coupled to the induction of insulin resistance. PMID- 16705065 TI - Mechanism of insulin's anabolic effect on muscle: measurements of muscle protein synthesis and breakdown using aminoacyl-tRNA and other surrogate measures. AB - Despite being an anabolic hormone in skeletal muscle, insulin's anticatabolic mechanism in humans remains controversial, with contradictory reports showing either stimulation of protein synthesis (PS) or inhibition of protein breakdown (PB) by insulin. Earlier measurements of muscle PS and PB in humans have relied on different surrogate measures of aminoacyl-tRNA and intracellular pools. We report that insulin's effect on muscle protein turnover using aminoacyl-tRNA as the precursor of PS and PB is calculated by mass balance of tracee amino acid (AA). We compared the results calculated from various surrogate measures. To determine the physiological role of insulin on muscle protein metabolism, we infused tracers of leucine and phenylalanine into 18 healthy subjects, and after 3 h, 10 subjects received a 4-h femoral arterial infusion of insulin (0.125 mUxkg(-1)xmin(-1)), while eight subjects continued with saline. Tracer-to-tracee ratios of leucine, phenylalanine, and ketoisocaproate were measured in the arterial and venous plasma, muscle tissue fluid, and AA-tRNA to calculate muscle PB and PS. Insulin infusion, unlike saline, significantly reduced the efflux of leucine and phenylalanine from muscle bed, based on various surrogate measures which agreed with those based on leucyl-tRNA (-28%), indicating a reduction in muscle PB (P < 0.02) without any significant effect on muscle PS. In conclusion, using AA-tRNA as the precursor pool, it is demonstrated that, in healthy humans in the postabsorptive state, insulin does not stimulate muscle protein synthesis and confirmed that insulin achieves muscle protein anabolism by inhibition of muscle protein breakdown. PMID- 16705066 TI - Myxedema pseudovolvulus. PMID- 16705067 TI - Unusual clinical presentation and possible rescue of a novel claudin-16 mutation. AB - CONTEXT: Familial hypomagnesemia with hypercalciuria and nephrocalcinosis (FHHNC) is caused by a dysfunction of Claudin-16 (CLDN16) and characterized by renal wasting of Mg(2+) and Ca(2+). OBJECTIVE: The objectives of this study were to study the clinical parameters in suspected FHHNC patients, identify mutations in the CLDN16 gene, and analyze molecular defects associated with the mutant protein. DESIGN, SETTING, AND PARTICIPANTS: CLDN16 genes from two siblings diagnosed with FHHNC were sequenced. Expression and characterization of the mutant protein in renal MDCK cells were studied. OUTCOME MEASURES: Standard urine and serum parameters to diagnose FHHNC were determined. Mutations in the CLDN16 gene were identified. The subcellular distribution of the mutant protein was analyzed by immunofluorescence microscopy. RESULTS: Urine and blood analysis showed signs typical for FHHNC. One patient, in addition, presented with hypocalcemic tetany, a phenomenon so far not described for FHHNC. Both siblings carry a novel mutation in CLDN16, Y207X. The review of medical records showed that hypocalcemia is not uncommon in the early childhood of FHHNC patients. Expressed in MDCK cells, the Y207X mutant is not detected at tight junctions but instead is found in lysosomes and, to a lesser extent, the endoplasmic reticulum. Surface expression can be rescued by inhibiting clathrin-mediated internalization. CONCLUSIONS: We propose that mutations in CLDN16 are considered in childhood hypocalcemia. CLDN16 Y207X is transiently delivered to the plasma membrane but not retained and is rapidly retrieved by internalization. Inhibitors of endocytosis may provide novel therapeutic strategies. PMID- 16705068 TI - Homozygous mutation of P450 side-chain cleavage enzyme gene (CYP11A1) in 46, XY patient with adrenal insufficiency, complete sex reversal, and agenesis of corpus callosum. AB - CONTEXT: The cholesterol side-chain cleavage enzyme catalyzes the conversion of cholesterol to pregnanalone in the first step of steroidogenic pathways. Defective enzyme activity leads to the deficiency of all steroid hormones, including progesterone, which is essential to sustain term pregnancy. RESULTS: We report a homozygous point mutation in the CYP11A1 gene in a 46, XY phenotypic female born at term to healthy heterozygous parents, presenting relatively late at the age of 1 yr 9 months with life-threatening adrenal insufficiency and complete sex reversal. She was found to have complete agenesis of corpus callosum. The mutation resulted in a single amino acid substitution: valine for alanine at position 359. The functional analysis of the mutant enzyme revealed markedly reduced enzyme activity, but about 11% residual activity was demonstrated. We explained the completion of pregnancy to term and the late presentation by a possible difference in the mutant enzyme activity in vivo and in vitro or by the residual mutant activity, which would have been enough to maintain pregnancy and viability of the patient. The clinical findings of nearly undetectable levels of steroid hormones at presentation are explained by the total disruption of steroidogenic cells later on, with recurrent ACTH stimulation leading to intramitochondrial cholesterol accumulation and cell death (a two-hit mechanism). CONCLUSION: This report of a homozygous mutation in CYP11A1 gene in a child with agenesis of corpus callosum shows that homozygous mutations in CYP11A1 gene can be compatible with term pregnancy and delayed presentation. PMID- 16705069 TI - Image in endocrinology: Localization of an occult adrenocorticotropin-secreting carcinoid tumor with somatostatin receptor scintigraphy using single-photon emission computed tomography/computed tomographic image scanning. PMID- 16705070 TI - Preserved sensitivity to beta2-adrenergic receptor agonists in patients with type 1 diabetes mellitus and hypoglycemia unawareness. AB - BACKGROUND AND OBJECTIVE: Use of beta(2)-adrenergic receptor agonists has been advocated for the treatment of hypoglycemia unawareness in type 1 diabetes. In vitro, however, hypoglycemia unawareness has been associated with reduced beta(2) adrenergic sensitivity. Therefore, in vivo sensitivity to beta(2)-adrenergic receptor agonist stimulation was compared between type 1 diabetic patients with and without hypoglycemia unawareness and nondiabetic controls. METHODS: Ten type 1 diabetic patients with hypoglycemia unawareness, 12 type 1 diabetic patients with intact hypoglycemic awareness, and 11 healthy controls were enrolled. beta(2)-Adrenergic sensitivity was determined by measuring the forearm vasodilator response to intraarterial infusion of salbutamol. Salbutamol was infused in six increasing doses ranging from 0.003 to 1.0 mug(1).min(-1).dl(-1). Forearm blood flow (FBF) was bilaterally measured by venous occlusion plethysmography. Diabetic patients received low-dose insulin before FBF measurements to ensure that experiments were carried out under normoglycemic conditions. RESULTS: At baseline, FBF was 1.9 +/- 0.3 ml(1).min(-1).dl(-1) in controls, 2.3 +/- 0.4 ml(1).min(-1).dl(-1) in patients with intact awareness, and 1.4 +/- 0.1 ml(1).min(-1).dl(-1) in patients with hypoglycemia unawareness (P = 0.048 vs. aware patients). In response to salbutamol, FBF increased 9.1-fold in controls, 8.0-fold in patients with intact awareness, and 10.7-fold in patients with hypoglycemia unawareness (P = NS). Heart rate increased in all groups due to systemic spillover of salbutamol but appeared blunted, considering a greater fall in mean arterial pressure in patients with hypoglycemia unawareness. CONCLUSIONS: Sensitivity to beta(2)-adrenergic receptor agonist stimulation is preserved in type 1 diabetic patients with hypoglycemia unawareness. PMID- 16705071 TI - Monosomy for the X-chromosome is associated with an atherogenic lipid profile. AB - CONTEXT AND OBJECTIVE: Men typically have a more atherogenic lipid profile than women characterized by higher low-density lipoprotein (LDL) cholesterol and triglyceride levels and reduced lipid particle size, contributing to a greater risk for coronary disease. To determine whether X-chromosomal gene dosage affects lipid metabolism independent of sex steroid effects, we compared lipid profiles in age- and body mass-matched young women with ovarian failure, differing only in X-chromosome dosage. DESIGN, SETTING, AND PATIENTS: Women with premature ovarian failure associated with monosomy X or Turner syndrome (TS, n = 118) were compared with women with 46,XX premature ovarian failure (n = 51) in an in-patient clinical research center unit at the National Institutes of Health. These women were normally on estrogen replacement treatment but discontinued the estrogen 2 wk before study. MAJOR OUTCOMES: Fasting lipid levels and nuclear magnetic resonance lipid particle profiles in the two study groups were the major outcomes. RESULTS: Average age and body mass were similar in the two groups of women, but LDL cholesterol (P = 0.001) and triglyceride levels (P = 0.0005) were higher in the TS group. Also among women with TS, average LDL particle size was reduced (P < 0.0001) and LDL particle concentration increased, with a 2-fold increase in the smallest particle categories (P < 0.0001). Whereas total high density lipoprotein cholesterol levels were similar, high-density lipoprotein particle size was significantly smaller in women with TS, compared with women with premature ovarian failure (P < 0.0001). CONCLUSIONS: Women with 45,X with ovarian failure exhibit a distinctly more atherogenic lipid profile than 46,XX women with ovarian failure, suggesting that the second X-chromosome contributes to a more salutary lipid profile in normal women, independent of sex steroid effects. PMID- 16705072 TI - Frequent finding of the androgen receptor A645D variant in normal population. AB - BACKGROUND: The androgen receptor A645D mutation has been described in one patient with ambiguous genitalia and one boy with normal phenotype. OBJECTIVE: Because of this phenotypic variation, we screened a cohort of men from the general population (n = 293) as well as men with the following disorders of the genital tract for the mutation: men with prostate cancer (n = 89), testicular cancer (n = 87), and infertility (n = 103). We also investigated the influence of the polymorphic CAG and GGN repeats on the phenotypic outcome. RESULTS: The A645D variant was found in three men from the general population (1.0%). These men did not differ regarding testosterone or LH concentrations, compared with the rest of this population. In addition, two men with prostate cancer (2.3%) and one infertile man (1.0%) presented with the mutation. No statistical differences in frequency were noted between the study groups, and none of these individuals had any genital malformations. All men who presented with the mutation carried an extraordinarily short GGN repeat of 10 base triplets in combination with long CAG repeats of 26-28 (average 27.3). In contrast, men with GGN=10, but CAG less than 26 did not have the A645D mutation. A single-nucleotide polymorphism analysis revealed that the A645D variant has emerged from the most common haplogroup in our population. CONCLUSIONS: We conclude that the A645D mutation, which is present in 1% of the general Swedish population, is linked to GGN10 and long CAG repeats. Its effect on androgen receptor function is currently unknown. PMID- 16705073 TI - Effects of testosterone supplementation on skeletal muscle fiber hypertrophy and satellite cells in community-dwelling older men. AB - OBJECTIVE: In this study, we determined the effects of graded doses of testosterone on muscle fiber cross-sectional area (CSA) and satellite cell number and replication in older men. PARTICIPANTS: Healthy men, 60-75 yr old, received a long-acting GnRH agonist to suppress endogenous testosterone production and 25, 50, 125, 300, or 600 mg testosterone enanthate im weekly for 20 wk. METHODS: Immunohistochemistry, light and confocal microscopy, and electron microscopy were used to perform fiber typing and quantitate myonuclear and satellite cell number in vastus lateralis biopsies, obtained before and after 20 wk of treatment. RESULTS: Testosterone administration in older men was associated with dose dependent increases in CSA of both types I and II fibers. Satellite cell number increased dose dependently at the three highest doses (3% at baseline vs. 6.2, 9.2, and 13.0% at 125, 300, and 600 mg doses, P < 0.05). Testosterone administration was associated with an increase in the number of proliferating cell nuclear antigen+ satellite cells (1.8% at baseline vs. 3.9, 7.5, and 13% at 125, 300, and 600 mg doses, P < 0.005). The expression of activated Notch, examined only in the 300-mg group (baseline, 2.3 vs. 9.0% after treatment, P < 0.005), increased in satellite cells after testosterone treatment. The expression of myogenin (baseline, 6.2 vs. 20.7% after treatment, P < 0.005), examined only in the 300-mg group, increased significantly in muscle fiber nuclei after testosterone treatment, but Numb expression did not change. CONCLUSIONS: Older men respond to graded doses of testosterone with a dose-dependent increase in muscle fiber CSA and satellite cell number. Testosterone-induced skeletal muscle hypertrophy in older men is associated with increased satellite cell replication and activation. PMID- 16705074 TI - Reduced expression of interleukin-11 and interleukin-6 in the periimplantation endometrium of excessive ovarian responders during in vitro fertilization treatment. AB - CONTEXT: Impaired implantation in assisted reproduction cycles with high serum estradiol (E2) concentrations may be related to abnormal endometrial functions. OBJECTIVE: The in vivo expression of T helper type 2 (Th2) cytokines in the periimplantation endometrium of infertile patients was compared between natural and stimulated cycles. INTERVENTIONS AND MAIN OUTCOME MEASURES: Uterine flushings and endometrial biopsies were collected 7 d after the LH surge in natural cycles or after human chorionic gonadotropin injection in stimulated cycles. Th2 cytokines were determined by immunolocalization and by ELISA. Natural cycles were in group A, whereas stimulated cycles with peak serum E2 of no more than 20,000 pmol/liter (moderate responders) and more than 20,000 pmol/liter (excessive responders) were classified as group B and group C, respectively. RESULTS: Higher E2 had a negative effect on IL-11 and IL-6 expression in the endometrium and IL 11 concentration in the uterine flushing. In endometrial biopsies, a significantly lower immunostaining of stromal IL-11 (P < 0.001) and glandular IL 6 (P < 0.05) was detected in group C compared with that of groups A and B. IL-11 concentration by ELISA was significantly lower in group C (P < 0.05). Endometrial leukemia inhibitory factor and IL-4 expression was similar in the three groups. In uterine flushings, a significantly higher percentage of women in group C had undetectable IL-11 and a lower IL-11 concentration (P < 0.01) compared with group A, whereas no difference in IL-6 concentration was noted in the three groups. CONCLUSION: Reduced expression of IL-11 and IL-6 in periimplantation endometrium may account for lower implantation in excessive responders. PMID- 16705075 TI - Elevated plasma adiponectin in humans with genetically defective insulin receptors. AB - CONTEXT: Adiponectin has been suggested to play a role in the etiopathogenesis of at least some forms of insulin resistance, in part based on a strong correlation between plasma levels of adiponectin and measures of insulin sensitivity. OBJECTIVE: The objective of the study was to establish whether this relationship is maintained at extreme levels of insulin resistance. DESIGN/SETTING: This was a cross-sectional study in a university teaching hospital of subjects recruited from the United Kingdom and the United States. PARTICIPANTS: Participants included 75 subjects with a range of syndromes of severe insulin resistance and 872 nondiabetic controls. OUTCOME MEASURES: Fasting plasma insulin, adiponectin, and leptin were measured. RESULTS: Unexpectedly, subjects with mutations in the insulin receptor, despite having the most severe degree of insulin resistance, had elevated plasma adiponectin [median 24.4 mg/liter; range 6.6-36.6 (normal adult range for body mass index 20 kg/m(2) = 3-19 mg/liter)], whereas all other subjects had low adiponectin levels (median 2.0 mg/liter; range 0.12-11.2). Plasma leptin in all but one subject with an insulin receptoropathy was low or undetectable [median 0.5 ng/ml; range 0-16: normal adult range for body mass index of < 25 kg/m(2) = 2.4-24.4 (female) and 0.4-8.3 ng/ml (male)]. CONCLUSIONS: We conclude that the relationship between plasma adiponectin and insulin sensitivity is complex and dependent on the precise etiology of defective insulin action and that the combination of high plasma adiponectin with low leptin may have clinical utility in patients with severe insulin resistance as a marker of the presence of a genetic defect in the insulin receptor. PMID- 16705076 TI - The N-terminal neurotensin fragment, NT1-11, inhibits cortisol secretion by human adrenocortical cells. AB - CONTEXT: Neurotensin (NT) modulates corticosteroid secretion from the mammalian adrenal gland. OBJECTIVE: The objective of this study was to investigate the possible involvement of NT in the control of cortisol secretion in the human adrenal gland. DESIGN: In vitro studies were conducted on cultured human adrenocortical cells. SETTING: This study was conducted in a university research laboratory. PATIENTS: Adrenal explants from patients undergoing expanded nephrectomy for kidney cancer were studied. MAIN OUTCOME MEASURE: Cortisol secretion from cultured adrenocortical cells was measured. RESULTS: NT1-11, the N terminal fragment of NT, dose-dependently inhibited basal and ACTH-stimulated cortisol production by human adrenocortical cells in primary culture. In contrast, NT had no influence on cortisol output at concentrations up to 10(-6) m. HPLC and RT-PCR analyses failed to detect any significant amounts of NT and NT mRNA, respectively, in adrenal extracts. Molecular and pharmacological studies were performed to determine the type of NT receptor involved in the corticostatic effect of NT1-11. RT-PCR analysis revealed the expression of NT receptor type (NTR) 3 mRNA but not NTR1 and NTR2 mRNAs in the human adrenal tissue. However, the pharmacological profile of the adrenal NT1-11 receptor was different from that of NTR3, indicating that this receptor type is not involved in the action of NT1-11 on corticosteroidogenesis. CONCLUSION: Our results indicate that NT1-11 may act as an endocrine factor to inhibit cortisol secretion through activation of a receptor distinct from the classical NTR1, NTR2, and NTR3. PMID- 16705077 TI - Effects of hormone replacement therapy type and route of administration on plasma matrix metalloproteinases and their tissue inhibitors in postmenopausal women. AB - BACKGROUND: Matrix metalloproteinases (MMPs) are implicated in numerous disease states including cardiovascular disease and cancer. Because recent studies have shown a detrimental effect of hormone replacement therapy on cardiovascular disease and breast cancer, we investigated whether there are any differences in the concentrations of MMPs and their tissue inhibitors (TIMPs) in women receiving various forms of postmenopausal therapy. MATERIAL AND METHODS: A total of 195 healthy postmenopausal women were assessed: 46 were taking tibolone, 47 were taking transdermal estradiol, 46 were taking conjugated equine estrogens (CEE), and 56 were not taking any menopausal therapy (CTR). Plasma levels of MMP-2 and 9 and TIMP-1 and TIMP-2 were measured by ELISA methods. RESULTS: MMP-9 levels were significantly higher in the CEE group in comparison with healthy women not receiving menopausal therapy (P < 0.05). In contrast, MMP-9 levels in the tibolone group were significantly lower than in any other group (P < 0.01, compared with transdermal estradiol and CTR, and P < 0.001, compared with CEE). MMP-9 to TIMP-1 ratio was also significantly higher in the CEE, compared with CTR (P < 0.05), and lower in the tibolone group (P < 0.01, compared with all groups). MMP-2 levels were higher in the CEE group, compared with healthy women not receiving any menopausal therapy, and women taking tibolone (P < 0.05). CONCLUSIONS: Our study demonstrates differential effects of various forms of postmenopausal therapy on serum levels of MMP-9 and MMP-2. It remains to be established whether these differences might be associated with differences in risks of cardiovascular disease and cancer in these women. PMID- 16705078 TI - Activation of dopamine D2 receptors lowers circadian leptin concentrations in obese women. AB - CONTEXT: Leptin release is regulated by factors other than fat mass alone. Previous observations provide indirect evidence for an inhibitory effect of dopaminergic neurotransmission on leptin secretion. This study was done to establish the effect of bromocriptine treatment on circadian plasma leptin concentrations in obese humans. OBJECTIVE: The objective of the study was to study the acute effects of bromocriptine (a D2R agonist) on circadian leptin levels in obese women, whereas body weight and caloric intake remained constant. DESIGN: This was a prospective, single-blind, crossover study (2004). SETTING: The study was conducted at a clinical research center. PARTICIPANTS: Eighteen healthy obese women (body mass index 33.2 +/- 0.6 kg/m(2)) were studied twice in the early follicular phase of their menstrual cycle. INTERVENTION(S): Treatment consisted of bromocriptine or placebo for 8 d. MAIN OUTCOME MEASURE(S): Blood was collected during 24 h at 20-min intervals for determination of leptin concentrations at the last day of medical treatment (bromocriptine or placebo). Mean 24-h serum concentrations were determined for insulin, glucose, free fatty acids, and triglycerides. RESULTS: Short-term treatment with bromocriptine reduced leptin concentration (placebo 33.6 +/- 2.5 vs. bromocriptine 30.5 +/- 2.5 ng/liter, P = 0.03). Free fatty acid concentrations were increased by treatment with bromocriptine. The increase of free fatty acids was inversely related with the decline of leptin levels. The decline of glucose, insulin, or prolactin concentrations in response to bromocriptine was not correlated with the reduction of leptin. CONCLUSION: Activation of dopamine D2 receptors by bromocriptine lowers circulating leptin levels in obese women, which suggests that dopaminergic neurotransmission is involved in the control of leptin release in humans. PMID- 16705079 TI - Spatial covariation of mutation and nonsynonymous substitution rates in vertebrate mitochondrial genomes. AB - Mitochondrial genomes encode fundamental subunits of the basic energy producing machinery of eukaryotic cells that are under strong functional constraint. Paradoxically, these genes evolve rapidly in general, and there is substantial variation in evolutionary rates among genes within genomes. In order to investigate spatial variation in selection intensity, we conducted tests of neutrality using ratios of synonymous to nonsynonymous substitutions (dN/dS = omega) on numerous protein gene segments from fishes and mammals. Values of omega were very low for nearly all genomic regions. However, values of both omega and dN varied in a clinal pattern with increasing distance from the light-strand origin of replication. Spatial heterogeneity of nonsynonymous substitution rates exhibits a significantly positive correlation with variation in mutation rates that are related to the mode of mitochondrial DNA replication. The finding that nonsynonymous substitution rates are proportional to mutation rates is expected if a majority of substitutions are selectively neutral or slightly deleterious. Spatial patterns of among-gene variation in nonsynonymous rates were highly similar between fishes and mammals, suggesting that forces governing mitochondrial gene evolution have remained relatively constant over 450 Myr of vertebrate evolution. Conservation of substitution patterns despite major shifts in thermal habit and metabolic demands among taxa implicates a conserved replication mechanism controlling relative mutation rates as a major determinant of mitochondrial protein evolution. PMID- 16705080 TI - Selective inhibition of plasma kallikrein protects brain from reperfusion injury. AB - We have studied the effect of DX-88, a selective recombinant inhibitor of human plasma kallikrein, in transient or permanent focal brain ischemia (with or without reperfusion, respectively) induced in C57BL/6 mice. Twenty-four hours after transient ischemia, DX-88 administered at the beginning of ischemia (pre) induced a dose-dependent reduction of ischemic volume that, at the dose of 30 microg/mouse, reached 49% of the volume of saline-treated mice. At the same dose, DX-88 was also able to reduce brain swelling to 32%. Mice treated with DX-88 pre had significantly lower general and focal deficit score. Fluoro-Jade staining, a marker for neuronal degeneration, showed that DX-88-treated mice had a reduction in the number of degenerating cells, compared with saline-treated mice. Seven days after transient ischemia, the DX-88 protective effect was still present. When the inhibitor was injected at the end of ischemia (post), it was still able to reduce ischemic volume, brain swelling, and neurological deficits. DX-88 efficacy was lost when the inhibitor was given 30 min after the beginning of reperfusion (1 h post) or when reperfusion was not present (permanent occlusion model). This study shows that DX-88 has a strong neuroprotective effect in the early phases of brain ischemia preventing reperfusion injury and indicates that inhibition of plasma kallikrein may be a useful tool in the strategy aimed at reducing the detrimental effects linked to reperfusion. PMID- 16705081 TI - Polymyxin B-conjugated alpha 2-macroglobulin as an adjunctive therapy to sepsis: Modes of action and impact on lethality. AB - A drug targeting both the inflammatory initiators (lipopolysaccharide; LPS) and mediators [tumor necrosis factor-alpha (TNF-alpha)] should have advantage over a "single-factor targeting strategy" in sepsis prevention trials. We have prepared conjugates of polymyxin B (PMB) and the cytokine binding protein alpha2 macroglobulin (A2M). The conjugate binds TNF-alpha as well as LPS as studied by electrophoresis and phase partitioning. Compared with free PMB, the conjugate is nontoxic to cells and does not affect the viability of human monocytes. The A2M PMB conjugate binds to the A2M receptor (CD91/low-density lipoprotein receptor related protein 1) with affinity similar to that of the nonmodified protein. Fluorescein isothiocyanate-labeled LPS in the presence of A2M-PMB is rapidly transported into fibroblasts for degradation via receptor-mediated endocytosis. In vitro, A2M-PMB demonstrated inhibition of LPS-induced secretion of TNF-alpha from isolated monocytes as well as in the whole blood assay. The efficacy of the drug was tested in mice after induction of acute inflammation (LPS model) and after induction of a polymicrobial sepsis by cecal ligation and puncture (CLP) model. Treatment of mice with A2M-PMB up to 250 microg/g body weight was not toxic to the animal. When the drug was administered 30 min before or 30 min after the LPS challenge, a survival rate of 90 and 70%, respectively, was obtained compared with the placebo control group (5%). A2M-PMB also protected mice after induction of polymicrobial sepsis when administered 30 min before CLP. These results support our hypothesis that A2M-PMB acts as a polyvalent drug to target different host mediators as well as sepsis inducer at the same time. PMID- 16705082 TI - Efficacy and the discriminative stimulus effects of negative GABAA modulators, or inverse agonists, in diazepam-treated rhesus monkeys. AB - In benzodiazepine (BZ)-dependent animals, the effects of negative GABA(A) modulators at BZ sites are not clearly related to differences in negative efficacy (i.e., inverse agonist activity). A flumazenil discriminative stimulus in diazepam (5.6 mg/kg/day)-treated rhesus monkeys was used to test the hypothesis that the effects of negative GABA(A) modulators at BZ sites do not vary as a function of efficacy in BZ-dependent animals. Negative GABA(A) modulators varying in efficacy were studied in combination with positive modulators acting at different modulatory sites (BZ, barbiturate, and neuroactive steroid sites). The negative modulators Ro 15-4513 (ethyl 8-azido-6-dihydro-5 methyl-6-oxo-4H-imidazo[1,5-alpha]-[1,4]benzodiazepine-3-carboxylate) and ethyl beta-carboline-3-carboxylate (beta-CCE) substituted for the flumazenil discriminative stimulus. Acute pretreatment with diazepam (3.2 and 10 mg/kg s.c., in addition to 5.6 mg/kg/day p.o.), pentobarbital (3.2 and 10 mg/kg), or pregnanolone (1 and 3.2 mg/kg) attenuated the flumazenil discriminative stimulus and also attenuated the flumazenil-like discriminative stimulus effects of Ro 15 4513 and beta-CCE. Attenuation of the discriminative stimulus effects of flumazenil, Ro 15-4513, and beta-CCE did not systematically vary as a function of negative efficacy. Compared with their discriminative stimulus effects in untreated monkeys discriminating midazolam, both pregnanolone and pentobarbital were relatively more potent than diazepam in attenuating the discriminative stimulus effects of flumazenil, Ro 15-4513, and beta-CCE in diazepam-treated monkeys. These results show that the discriminative stimulus effects of BZ-site neutral and negative modulators are not different in BZ-dependent animals trained to discriminate flumazenil, and extend the results of a previous study showing that positive modulators acting at non-BZ sites are especially potent in attenuating the effects of flumazenil in diazepam-treated monkeys (i.e., diazepam withdrawal). PMID- 16705083 TI - Gene duplication and functional divergence of the zebrafish insulin-like growth factor 1 receptors. AB - Insulin-like growth factor (IGF) 1 receptor (IGF1R)-mediated signaling plays key roles in growth, development, and physiology. Recent studies have shown that there are two distinct ig f1r genes in zebrafish, termed ig f1ra and ig f1rb. In this study, we tested the hypothesis that zebrafish ig f1ra and ig f1rb resulted from a gene duplication event at the ig f1r locus and that this has led to their functional divergence. The genomic structures of zebrafish ig f1ra and ig f1rb were determined and their loci mapped. While zebrafish ig f1ra has 21 exons and is located on linkage group (LG) 18, zebrafish ig f1rb has 22 exons and mapped to LG 7. There is a strong syntenic relationship between the two zebrafish genes and the human IG F1R gene. Using a MO-based loss-of-function approach, we show that both Igf1ra and Igf1rb are required for zebrafish embryo viability and proper growth and development. Although Igf1ra and Igf1rb demonstrated a large degree of functional overlap with regard to cell differentiation in the developing eye, inner ear, heart, and muscle, they also exhibited functional distinction involving a greater requirement for Igf1rb in spontaneous muscle contractility. These findings suggest that the duplicated zebrafish ig f1r genes play largely overlapping but not identical functional roles in early development and provide novel insight into the functional evolution of the IGF1R/insulin receptor gene family. PMID- 16705084 TI - Do CD8 effector cells need IL-7R expression to become resting memory cells? AB - The role for IL-7R expression in the differentiation of effector T cells into resting memory remains controversial. Here, using a conditional IL-7R transgenic model, we were able to test directly whether CD8 effector T cells require IL-7R expression for their differentiation into resting memory cells. In the absence of IL-7R expression, effector cells transferred into "full" hosts underwent a protracted and unremitting contraction compared with IL-7R-expressing control cells and were unable to develop into long-term resting memory cells. Surprisingly, when the same effector cells were transferred into empty T-cell deficient hosts, they could generate long-lived fully functional resting memory cells independently of IL-7R expression. Formation of these latter cells was found to be dependent on IL-15, because the same IL-7R-deficient effector cells were rapidly lost from IL-15-deficient hosts, having a half-life of less than 40 hours. Therefore, our data suggest that, under physiological conditions, both IL 7 and IL-15 synergize to promote the formation of memory cells directly by limiting the contraction of effectors that occurs following an immune response and that reexpression of IL-7R is a key checkpoint in the regulation of this process. PMID- 16705085 TI - Analysis of fibrinogen variants at gamma387Ile shows that the side chain of gamma387 and the tertiary structure of the gammaC-terminal tail are important not only for assembly and secretion of fibrinogen but also for lateral aggregation of protofibrils and XIIIa-catalyzed gamma-gamma dimer formation. AB - To examine the role of fibrinogen gamma-chain residue 387Ile in the assembly and secretion of this multichain protein, we synthesized a series of variants with substitution at gamma387 by Arg, Leu, Met, Ala, or Asp. Only the variant gamma387Asp showed impaired synthesis in the cells and very low secretion into the medium. In addition, we performed thrombin-catalyzed fibrin polymerization and factor (F) XIIIa-catalyzed cross-linking of the gamma-chain for 4 variants. The degree of lateral aggregation of protofibrils into fibrin fibers was slightly reduced for gamma387Arg and Ala, and moderately reduced for gamma387Leu and Met. Although the FXIIIa-catalyzed cross-linking for all of the variants was slower than that for gamma387Ile, that of gamma387Arg was much more markedly impaired than that of the others. In summary, our studies demonstrated that the specific residue at gamma387 or the conformation of gamma388-411 residues, but not the length of the gammaC tail, is critical for fibrinogen assembly and subsequent secretion. Moreover, this residue or the conformation is also important for not only the lateral aggregation of fibrin polymers but also the FXIIIa-catalyzed cross-linking of the gamma-chain. Interestingly, our results clearly indicate that the conformations critical for these 2 functions are different from each other. PMID- 16705086 TI - The epitope recognized by rituximab. AB - Rituximab is a monoclonal antibody widely used in the treatment of malignant lymphoma and autoimmunity. Its epitope within the B-cell antigen CD20 is largely unknown. We used phage display libraries to select peptides binding to rituximab. Enriched peptides showed 2 sequence patterns: one motif (CALMIANSC) is related to (170)ANPS(173) within CD20, while another motif (WEWTI) may mimic the CD20 segment (182)YCYSI(185). Phages displaying either motif specifically bound rituximab. Binding to rituximab by the CD20 peptides ANPS and YCYSI was weak when used separately and enhanced when both peptides were linked. Recombinant CD20 extracellular loop proteins blocked binding of the selected CWWEWTIGC phage to rituximab, suggesting that CWWEWTIGC mimics the epitope. Blocking capacity was strongly reduced upon mutation of the CD20 strings ANPS or YCYSI. We conclude that rituximab binds a discontinuous epitope in CD20, comprised of (170)ANPS(173) and (182)YCYSI(185), with both strings brought in steric proximity by a disulfide bridge between C(167) and C(183). PMID- 16705087 TI - Cell death provoked by loss of interleukin-3 signaling is independent of Bad, Bim, and PI3 kinase, but depends in part on Puma. AB - Growth and survival of hematopoietic cells is regulated by growth factors and cytokines, such as interleukin 3 (IL-3). When cytokine is removed, cells dependent on IL-3 kill themselves by a mechanism that is inhibited by overexpression of Bcl-2 and is likely to be mediated by proapoptotic Bcl-2 family members. Bad and Bim are 2 such BH3-only Bcl-2 family members that have been implicated as key initiators in apoptosis following growth factor withdrawal, particularly in IL-3-dependent cells. To test the role of Bad, Bim, and other proapoptotic Bcl-2 family members in IL-3 withdrawal-induced apoptosis, we generated IL-3-dependent cell lines from mice lacking the genes for Bad, Bim, Puma, both Bad and Bim, and both Bax and Bak. Surprisingly, Bad was not required for cell death following IL-3 withdrawal, suggesting changes to phosphorylation of Bad play only a minor role in apoptosis in this system. Deletion of Bim also had no effect, but cells lacking Puma survived and formed colonies when IL-3 was restored. Inhibition of the PI3 kinase pathway promoted apoptosis in the presence or absence of IL-3 and did not require Bad, Bim, or Puma, suggesting IL-3 receptor survival signals and PI3 kinase survival signals are independent. PMID- 16705088 TI - Opposing roles of blood myeloid and plasmacytoid dendritic cells in HIV-1 infection of T cells: transmission facilitation versus replication inhibition. AB - CD11c(+) myeloid dendritic cells (MDCs) and CD11c(-) CD123(+) plasmacytoid DCs (PDCs) have been identified as main human DC subsets. MDCs are professional antigen-presenting cells for T cells, and include Langerhans cells, dermal DCs, and interstitial DCs. They have been associated with HIV-1 capture and sexual transmission, whereas PDCs play an important role in the innate immune responses to different types of viruses, including HIV-1. To compare the influence of MDCs and PDCs on HIV-1 infection of T cells, we isolated donor-matched MDCs and PDCs from peripheral blood, activated them by adding different maturation-inducing compounds, and cocultured them with T cells and HIV-1. We found that MDCs enhance HIV-1 infection through capture of the virus and subsequent transmission to T cells, and that differently matured MDC subsets have different HIV-1 transmission efficiencies. These differences were not due to soluble factors, viral capture differences, or the expression of integrins ICAM-1, -2, -3, or LFA-1. In contrast, regardless of their state of maturation, PDCs inhibit HIV-1 replication in T cells through the secretion of IFNalpha and an additional, unidentified small molecule. This study shows that the 2 main types of DCs have opposing roles in HIV-1 infection of T cells. PMID- 16705090 TI - Integration of global SNP-based mapping and expression arrays reveals key regions, mechanisms, and genes important in the pathogenesis of multiple myeloma. AB - Multiple myeloma is characterized by genomic alterations frequently involving gains and losses of chromosomes. Single nucleotide polymorphism (SNP)-based mapping arrays allow the identification of copy number changes at the sub megabase level and the identification of loss of heterozygosity (LOH) due to monosomy and uniparental disomy (UPD). We have found that SNP-based mapping array data and fluorescence in situ hybridization (FISH) copy number data correlated well, making the technique robust as a tool to investigate myeloma genomics. The most frequently identified alterations are located at 1p, 1q, 6q, 8p, 13, and 16q. LOH is found in these large regions and also in smaller regions throughout the genome with a median size of 1 Mb. We have identified that UPD is prevalent in myeloma and occurs through a number of mechanisms including mitotic nondisjunction and mitotic recombination. For the first time in myeloma, integration of mapping and expression data has allowed us to reduce the complexity of standard gene expression data and identify candidate genes important in both the transition from normal to monoclonal gammopathy of unknown significance (MGUS) to myeloma and in different subgroups within myeloma. We have documented these genes, providing a focus for further studies to identify and characterize those that are key in the pathogenesis of myeloma. PMID- 16705091 TI - Fine mapping of quantitative trait nucleotides underlying thrombin-activatable fibrinolysis inhibitor antigen levels by a transethnic study. AB - Recent studies revisiting the association between plasma thrombin-activatable fibrinolysis inhibitor (TAFI) Ag levels and polymorphisms of the CPB2 gene (coding for TAFI) suggested that TAFI Ag levels were influenced by 2 major quantitative trait nucleotides (QTNs) in European whites. However, the strong linkage disequilibrium (LD) between CPB2 polymorphisms in European whites did not allow one to distinguish which polymorphisms could be the putative QTNs. To get a better insight into the identification of QTNs, a transethnic haplotype analysis contrasting 2 populations of African and European subjects was performed using 13 CPB2 polymorphisms. Results of the haplotype analyses suggested that 3 QTNs had independent effects and explained about 15% of the TAFI variability, consistently in the 2 populations. The lower LD observed in the African population enabled us to identify the 1583T>A SNP located in 3'UTR as one of these QTNs, whereas the 2599C>G and -2345--2344insG SNPs located in the 5' region might be the 2 other QTNs. A phylogenetic study suggested that these 3 polymorphisms occurred before the period of migration "out of Africa." Although this transethnic comparison contributed to better map the putative CPB2 QTNs, further studies are required to clarify the role of the promoter region. PMID- 16705089 TI - Frequent gain of chromosome band 1q21 in plasma-cell dyscrasias detected by fluorescence in situ hybridization: incidence increases from MGUS to relapsed myeloma and is related to prognosis and disease progression following tandem stem cell transplantation. AB - Using fluorescence in situ hybridization we investigated amplification of chromosome band 1q21 (Amp1q21) in more than 500 untreated patients with monoclonal gammopathy of undetermined significance (MGUS; n = 14), smoldering multiple myeloma (SMM; n = 31), and newly diagnosed MM (n = 479) as well as 45 with relapsed MM. The frequency of Amp1q21 was 0% in MGUS, 45% in SMM, 43% in newly diagnosed MM, and 72% in relapsed MM (newly diagnosed versus relapsed MM, P < .001). Amp1q21 was detected in 10 of 12 patients whose disease evolved to active MM compared with 4 of 19 who remained with SMM (P < .001). Patients with newly diagnosed MM with Amp1q21 had inferior 5-year event-free/overall survival compared with those lacking Amp1q21 (38%/52% versus 62%/78%, both P < .001). Thalidomide improved 5-year EFS in patients lacking Amp1q21 but not in those with Amp1q21 (P = .004). Multivariate analysis including other major predictors revealed that Amp1q21 was an independent poor prognostic factor. Relapsed patients who had Amp1q21 at relapse had inferior 5-year postrelapse survival compared with those lacking Amp1q21 at relapse (15% versus 53%, P = .027). The proportion of cells with Amp1q21 and the copy number of 1q21 tended to increase at relapse compared with diagnosis. Our data suggest that Amp1q21 is associated with both disease progression and poor prognosis. PMID- 16705092 TI - Transforming growth factor-beta1 regulates macrophage migration via RhoA. AB - Brief treatment with transforming growth factor (TGF)-beta1 stimulated the migration of macrophages, whereas long-term exposure decreased their migration. Cell migration stimulated by TGF-beta1 was markedly inhibited by 10 mug/mL Tat-C3 exoenzyme. TGF-beta1 increased mRNA and protein levels of macrophage inflammatory protein (MIP)-1alpha in the initial period, and these effects also were inhibited by 10 mug/mL Tat-C3 and a dominant-negative (DN)-RhoA (N19RhoA). Cycloheximide, actinomycin D, and antibodies against MIP-1alpha and monocyte chemoattractant protein-1 (MCP-1) abolished the stimulation of cell migration by TGF-beta1. These findings suggest that migration of these cells is regulated directly and indirectly via the expression of chemokines such as MIP-1alpha and MCP-1 mediated by RhoA in response to TGF-beta1. TGF-beta1 activated RhoA in the initial period, and thereafter inactivated them, suggesting that the inactivation of RhoA may be the cause of the reduced cell migration in response to TGF-beta1 at later times. We therefore attempted to elucidate the molecular mechanism of the inactivation of RhoA by TGF-beta1. First, TGF-beta1 phosphorylated RhoA via protein kinase A, leading to inactivation of RhoA. Second, wild-type p190 Rho GTPase activating protein (p190RhoGAP) reduced and DN-p190RhoGAP reversed the reduction of cell migration induced by TGF-beta, suggesting that it inactivated RhoA via p190 Rho GAP. PMID- 16705094 TI - The cover. Under a Cloud. PMID- 16705095 TI - A piece of my mind. Shut-ins. PMID- 16705096 TI - US stem cell research lagging. PMID- 16705097 TI - New markers may help predict prostate cancer relapse risk. PMID- 16705093 TI - Lyn kinase promotes erythroblast expansion and late-stage development. AB - Lyn kinase is known to modulate the formation and function of B cells, monocytes, and mast cells. However, Lyn-/- mice also develop erythrosplenomegaly, and cases for both negative and positive erythropoietic actions of Lyn recently have been outlined. In phenylhydrazine-treated Lyn-/- mice, extramedullary splenic erythropoiesis was hyperactivated, but this did not lead to accelerated recovery from anemia. Furthermore, ex vivo analyses of the development of bone marrow derived Lyn-/- erythroblasts in unique primary culture systems indicated positive roles for Lyn at 2 stages. Late-stage Lyn-/- erythroblasts exhibited deficit Ter119(pos) cell formation, and this was paralleled by increased apoptosis (and decreased Bcl-xL expression). During early development, Lyn-/- erythroblasts accumulated at a Kit(pos)CD71(high) stage, possessed decreased proliferative capacity, and were attenuated in entering an apparent G1/S cell-cycle phase. In proposed compensatory responses, Lyn-/- erythroblasts expressed increased levels of activated Akt and p60-Src and decreased levels of death-associated protein kinase-2. Stat5 activation and Bcl-xL expression, in contrast, were significantly decreased in keeping with decreased survival and developmental potentials. Lyn, therefore, is proposed to function via erythroid cell-intrinsic mechanisms to promote progenitor cell expansion beyond a Kit(pos)CD71(high) stage and to support subsequent late-stage development. PMID- 16705098 TI - Group to revamp vector-control methods. PMID- 16705099 TI - Nature's glue may have medical uses. PMID- 16705100 TI - Prevention of atrial fibrillation following cardiac surgery. PMID- 16705101 TI - Prevention of atrial fibrillation following cardiac surgery. PMID- 16705102 TI - Prevention of atrial fibrillation following cardiac surgery. PMID- 16705103 TI - Cocktails for two. PMID- 16705104 TI - Psychiatric implications of psoriasis. PMID- 16705105 TI - Cocktails for two. PMID- 16705106 TI - Effect of hypobaric hypoxia, simulating conditions during long-haul air travel, on coagulation, fibrinolysis, platelet function, and endothelial activation. AB - CONTEXT: The link between long-haul air travel and venous thromboembolism is the subject of continuing debate. It remains unclear whether the reduced cabin pressure and oxygen tension in the airplane cabin create an increased risk compared with seated immobility at ground level. OBJECTIVE: To determine whether hypobaric hypoxia, which may be encountered during air travel, activates hemostasis. DESIGN, SETTING, AND PARTICIPANTS: A single-blind, crossover study, performed in a hypobaric chamber, to assess the effect of an 8-hour seated exposure to hypobaric hypoxia on hemostasis in 73 healthy volunteers, which was conducted in the United Kingdom from September 2003 to November 2005. Participants were screened for factor V Leiden G1691A and prothrombin G20210A mutation and were excluded if they tested positive. Blood was drawn before and after exposure to assess activation of hemostasis. INTERVENTIONS: Individuals were exposed alternately (> or =1 week apart) to hypobaric hypoxia, similar to the conditions of reduced cabin pressure during commercial air travel (equivalent to atmospheric pressure at an altitude of 2438 m), and normobaric normoxia (control condition; equivalent to atmospheric conditions at ground level, circa 70 m above sea level). MAIN OUTCOME MEASURES: Comparative changes in markers of coagulation activation, fibrinolysis, platelet activation, and endothelial cell activation. RESULTS: Changes were observed in some hemostatic markers during the normobaric exposure, attributed to prolonged sitting and circadian variation. However, there were no significant differences between the changes in the hypobaric and the normobaric exposures. For example, the median difference in change between the hypobaric and normobaric exposure was 0 ng/mL for thrombin antithrombin complex (95% CI, -0.30 to 0.30 ng/mL); -0.02 [corrected] nmol/L for prothrombin fragment 1 + 2 (95% CI, -0.03 to 0.01 nmol/L); 1.38 ng/mL for D-dimer (95% CI, -3.63 to 9.72 ng/mL); and -2.00% for endogenous thrombin potential (95% CI, -4.00% to 1.00%). CONCLUSION: Our findings do not support the hypothesis that hypobaric hypoxia, of the degree that might be encountered during long-haul air travel, is associated with prothrombotic alterations in the hemostatic system in healthy individuals at low risk of venous thromboembolism. PMID- 16705107 TI - Effect of policosanol on lipid levels among patients with hypercholesterolemia or combined hyperlipidemia: a randomized controlled trial. AB - CONTEXT: Policosanol is a natural substance derived from sugar cane that is advertised for its lipid-lowering effects as a nonprescription drug. More than 80 placebo-controlled or comparative trials, performed mostly by a single research institute, suggest that policosanol at doses of 5 to 40 mg/d has lipoprotein lowering effects comparable with statins. OBJECTIVES: To determine the lipoprotein-lowering effects of Cuban sugar cane-derived policosanol and to establish, if effective, dose-dependency up to 80 mg/d in patients with hypercholesterolemia or combined hyperlipidemia. DESIGN, SETTING, AND PARTICIPANTS: A multicenter (lipid outpatient clinics and general practitioners in Germany), randomized, double-blind, placebo-controlled, parallel-group trial conducted from September 29, 2000, to May 10, 2001, of patients with hypercholesterolemia or combined hyperlipidemia having baseline low-density lipoprotein cholesterol (LDL-C) levels of at least 150 mg/dL (> or =3.88 mmol/L) and either no or 1 cardiovascular risk factor other than known coronary heart disease, or baseline LDL-C levels of between 150 and 189 mg/dL (3.88-4.89 mmol/L) and 2 or more risk factors. INTERVENTIONS: Open-label 6-week placebo and diet run in phase followed by a double-blind 12-week treatment phase after randomization to 5 groups: 10, 20, 40, or 80 mg/d of policosanol or placebo. MAIN OUTCOME MEASURE: The percentage change of LDL-C, with changes in other lipoproteins as secondary outcome measures. RESULTS: A total of 143 patients were randomized to 5 equal groups and were analyzed on an intention-to-treat basis. In none of the 5 treatment groups did LDL-C levels decrease more than 10% from baseline. No statistically significant difference between policosanol and placebo was observed. A nonparametric test analyzing dose-dependency yielded nonsignificant results. In none of the secondary outcome measures, namely total cholesterol, high-density lipoprotein cholesterol (HDL-C), very low-density lipoprotein cholesterol, triglycerides, lipoprotein(a), and ratio of total or LDL-C to HDL-C, were there any significant effects of policosanol. Policosanol was tolerated well without serious adverse events. CONCLUSION: In patients with hypercholesterolemia or combined hyperlipidemia, the sugar cane-derived policosanol in usual and high doses does not demonstrate a reduction in lipid levels beyond placebo. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT00288483. PMID- 16705108 TI - Reported outcomes in major cardiovascular clinical trials funded by for-profit and not-for-profit organizations: 2000-2005. AB - CONTEXT: In surveys based on data available prior to 2000, clinical trials funded by for-profit organizations appeared more likely to report positive findings than those funded by not-for-profit organizations. Whether this situation has changed over the past 5 years or whether similar effects are present among jointly funded trials is unknown. OBJECTIVE: To determine in contemporary randomized cardiovascular trials the association between funding source and the likelihood of reporting positive findings. DESIGN: We reviewed 324 consecutive superiority trials of cardiovascular medicine published between January 1, 2000, and July 30, 2005, in JAMA, The Lancet, and the New England Journal of Medicine. MAIN OUTCOME MEASURE: The proportion of trials favoring newer treatments over the standard of care was evaluated by funding source. RESULTS: Of the 324 superiority trials, 21 cited no funding source. Of the 104 trials funded solely by not-for-profit organizations, 51 (49%) reported evidence significantly favoring newer treatments over the standard of care, whereas 53 (51%) did not (P = .80). By contrast, 92 (67.2%) of 137 trials funded solely by for-profit organizations favored newer treatments over standard of care (P<.001). Among 62 jointly funded trials, 35 (56.5%), an intermediate proportion, favored newer treatments. For 205 randomized trials evaluating drugs, the proportions favoring newer treatments were 39.5%, not-for-profit; 54.4%, jointly funded; and 65.5%, for-profit trials (P for trend across groups = .002). For the 39 randomized trials evaluating cardiovascular devices, the proportions favoring newer treatments were 50.0%, not-for-profit; 69.2%, jointly funded; and 82.4%, for-profit trials (P for trend across groups = .07). Regardless of funding source, trials using surrogate end points, such as quantitative angiography, intravascular ultrasound, plasma biomarkers, and functional measures were more likely to report positive findings (67%) than trials using clinical end points (54.1%; P = .02). CONCLUSIONS: Recent cardiovascular trials funded by for-profit organizations are more likely to report positive findings than trials funded by not-for-profit organizations, as are trials using surrogate rather than clinical end points. Trials jointly funded by not-for-profit and for-profit organizations appear to report positive findings at a rate approximately midway between rates observed in trials supported solely by one or the other of these entities. PMID- 16705109 TI - Anti-TNF antibody therapy in rheumatoid arthritis and the risk of serious infections and malignancies: systematic review and meta-analysis of rare harmful effects in randomized controlled trials. AB - CONTEXT: Tumor necrosis factor (TNF) plays an important role in host defense and tumor growth control. Therefore, anti-TNF antibody therapies may increase the risk of serious infections and malignancies. OBJECTIVE: To assess the extent to which anti-TNF antibody therapies may increase the risk of serious infections and malignancies in patients with rheumatoid arthritis by performing a meta-analysis to derive estimates of sparse harmful events occurring in randomized trials of anti-TNF therapy. DATA SOURCES: A systematic literature search of EMBASE, MEDLINE, Cochrane Library, and electronic abstract databases of the annual scientific meetings of both the European League Against Rheumatism and the American College of Rheumatology was conducted through December 2005. This search was complemented with interviews of the manufacturers of the 2 licensed anti-TNF antibodies. STUDY SELECTION: We included randomized, placebo-controlled trials of the 2 licensed anti-TNF antibodies (infliximab and adalimumab) used for 12 weeks or more in patients with rheumatoid arthritis. Nine trials met our inclusion criteria, including 3493 patients who received anti-TNF antibody treatment and 1512 patients who received placebo. DATA EXTRACTION: Data on study characteristics to assess study quality and intention-to-treat data for serious infections and malignancies were abstracted. Published information from the trials was supplemented by direct contact between principal investigators and industry sponsors. DATA SYNTHESIS: We calculated a pooled odds ratio (Mantel Haenszel methods with a continuity correction designed for sparse data) for malignancies and serious infections (infection that requires antimicrobial therapy and/or hospitalization) in anti-TNF-treated patients vs placebo patients. We estimated effects for high and low doses separately. The pooled odds ratio for malignancy was 3.3 (95% confidence interval [CI], 1.2-9.1) and for serious infection was 2.0 (95% CI, 1.3-3.1). Malignancies were significantly more common in patients treated with higher doses compared with patients who received lower doses of anti-TNF antibodies. For patients treated with anti-TNF antibodies in the included trials, the number needed to harm was 154 (95% CI, 91-500) for 1 additional malignancy within a treatment period of 6 to 12 months. For serious infections, the number needed to harm was 59 (95% CI, 39-125) within a treatment period of 3 to 12 months. CONCLUSIONS: There is evidence of an increased risk of serious infections and a dose-dependent increased risk of malignancies in patients with rheumatoid arthritis treated with anti-TNF antibody therapy. The formal meta-analysis with pooled sparse adverse events data from randomized controlled trials serves as a tool to assess harmful drug effects. PMID- 16705110 TI - Spontaneous spinal cerebrospinal fluid leaks and intracranial hypotension. AB - CONTEXT: Spontaneous intracranial hypotension is caused by spontaneous spinal cerebrospinal fluid (CSF) leaks and is known for causing orthostatic headaches. It is an important cause of new headaches in young and middle-aged individuals, but initial misdiagnosis is common. OBJECTIVE: To summarize existing evidence regarding the epidemiology, pathophysiology, diagnosis, and management of spontaneous spinal CSF leaks and intracranial hypotension. EVIDENCE ACQUISITION: MEDLINE (1966-2005) and OLDMEDLINE (1950-1965) were searched using the terms intracranial hypotension, CSF leak, low pressure headache, and CSF hypovolemia. Reference lists of these articles and ongoing investigations in this area were used as well. EVIDENCE SYNTHESIS: Spontaneous intracranial hypotension is caused by single or multiple spinal CSF leaks. The incidence has been estimated at 5 per 100,000 per year, with a peak around age 40 years. Women are affected more commonly than men. Mechanical factors combine with an underlying connective tissue disorder to cause the CSF leaks. An orthostatic headache is the prototypical manifestation but other headache patterns occur as well, and associated symptoms are common. Typical magnetic resonance imaging findings include subdural fluid collections, enhancement of the pachymeninges, engorgement of venous structures, pituitary hyperemia, and sagging of the brain (mnemonic: SEEPS). Myelography is the study of choice to identify the spinal CSF leak. Treatments include bed rest, epidural blood patching, percutaneous placement of fibrin sealant, and surgical CSF leak repair, but outcomes have been poorly studied and no management strategies have been studied in properly controlled randomized trials. CONCLUSIONS: Spontaneous intracranial hypotension is not rare but it remains underdiagnosed. The spectrum of clinical and radiographic manifestations is varied, with diagnosis largely based on clinical suspicion, cranial magnetic resonance imaging, and myelography. Numerous treatment options are available, but much remains to be learned about this disorder. PMID- 16705111 TI - How thrombogenic is hypoxia? PMID- 16705112 TI - JAMA patient page. Headaches. PMID- 16705113 TI - Pak up your breast tumor--and grow! PMID- 16705114 TI - Caring for older women with breast cancer: can observational research fill the clinical trial gap? PMID- 16705115 TI - Scientists are searching the seas for cancer drugs. PMID- 16705116 TI - Stat bite: Causes of lung cancer in nonsmokers. PMID- 16705117 TI - Attention turns to lung cancer in nonsmokers. PMID- 16705118 TI - Celecoxib shown effective in preventing colon polyps. PMID- 16705119 TI - World Health Organization focuses on antitobacco efforts in developing nations through treaty. PMID- 16705120 TI - New research suggests access, genetic differences play role in high minority cancer death rate. PMID- 16705121 TI - Association between Pak1 expression and subcellular localization and tamoxifen resistance in breast cancer patients. AB - BACKGROUND: p21-activated kinase 1 (Pak1) phosphorylates many proteins in both normal and transformed cells. Its ability to phosphorylate and thereby activate the estrogen receptor alpha (ERalpha) potentially limits the effectiveness of antiestrogen treatment in breast cancer. Here we studied associations between Pak1 expression and subcellular localization in tumor cells and tamoxifen resistance. METHODS: Pak1 protein expression was evaluated in 403 primary breast tumors from premenopausal patients who had been randomly assigned to 2 years of adjuvant tamoxifen or no treatment. Tamoxifen response was evaluated by comparing recurrence-free survival in relation to Pak1 and ERalpha expression in untreated versus tamoxifen-treated patients. Tamoxifen responsiveness of human MCF-7 breast cancer cells that inducibly expressed constitutively active Pak1 or that transiently overexpressed wild-type Pak1 (Wt-Pak1) or Pak1 that lacked functional nuclear localization signals (Pak1DeltaNLS) was evaluated by analyzing cyclin D1 promoter activation and protein levels as markers for ERalpha activation. The response to tamoxifen in relation to Pak1 expression was analyzed in naturally tamoxifen-resistant Ishikawa human endometrial cancer cells. All statistical tests were two-sided. RESULTS: Among patients who had ERalpha-positive tumors with low Pak1 expression, those treated with tamoxifen had better recurrence-free survival than those who received no treatment (hazard ratio [HR] = 0.502, 95% confidence interval [CI] = 0.331 to 0.762; P = .001) whereas there was no difference in recurrence-free survival between treatment groups for patients whose tumors had high cytoplasmic (HR = 0.893, 95% CI = 0.420 to 1.901; P = .769) or any nuclear Pak1 expression (HR = 0.955, 95% CI = 0.405 to 2.250; P = .916). In MCF-7 cells, overexpression of Wt-Pak1, but not of Pak1DeltaNLS, compromised tamoxifen response by stimulating cyclin D1 expression. Treatment of Ishikawa cells with tamoxifen led to an increase in the amount of nuclear Pak1 and Pak1 kinase activity, suggesting that tamoxifen, to some extent, regulates Pak1 expression. CONCLUSIONS: Our data support a role for Pak1, particular Pak1 localized to the nucleus, in ERalpha signaling and in tamoxifen resistance. PMID- 16705122 TI - Effectiveness of radiation therapy for older women with early breast cancer. AB - BACKGROUND: Recent clinical trials have questioned the necessity of breast radiation therapy for older women with early breast cancer. However, the effectiveness of radiation therapy for older women in the community setting has not been addressed. METHODS: We used the Surveillance, Epidemiology, and End Results (SEER)-Medicare database from January 1, 1992, through December 31, 1999, to identify 8724 women aged 70 years or older treated with conservative surgery for small, lymph node-negative, estrogen receptor-positive (or unknown receptor status) breast cancer. We used a proportional hazards model to test whether radiation therapy was associated with a lower risk of a combined outcome, defined as a second ipsilateral breast cancer reported by SEER and/or a subsequent mastectomy reported by Medicare claims. All statistical tests were two-sided. RESULTS: Radiation therapy, compared with no radiation therapy, was associated with a lower risk of the combined outcome (hazard ratio = 0.19, 95% confidence interval = 0.14 to 0.28). Radiation therapy was associated with an absolute risk reduction of 4.0 events per 100 women at 5 years (i.e., from 5.1 events without radiation therapy to 1.1 with radiation therapy) and 5.7 events per 100 persons at 8 years (i.e., from 8.0 events without radiation therapy to 2.3 with radiation therapy) (P < .001, log-rank test). Radiation therapy was most likely to benefit those aged 70-79 years without comorbidity (number needed to treat [NNT] to prevent one event = 21 to 22 patients) and was least likely to benefit those aged 80 years or older with moderate to severe comorbidity (NNT = 61 to 125 patients). CONCLUSION: For older women with early breast cancer, radiation therapy was associated with a lower risk of a second ipsilateral breast cancer and subsequent mastectomy. Patients aged 70-79 years with minimal comorbidity were the most likely to benefit, and older patients with substantial comorbidity were least likely to benefit. PMID- 16705123 TI - Lung cancer death rates in lifelong nonsmokers. AB - BACKGROUND: Few studies have directly measured the age-, sex-, and race-specific risks of lung cancer incidence and mortality among never tobacco smokers. Such data are needed to quantify the risks associated with smoking and to understand racial and sex disparities and temporal trends that are due to factors other than active smoking. METHODS: We measured age-, sex-, and race-specific rates (per 100,000 person-years at risk) of death from lung cancer among more than 940,000 adults who reported no history of smoking at enrollment in either of two large American Cancer Society Cancer Prevention Study cohorts during 1959-1972 (CPS-I) and 1982-2000 (CPS-II). We compared lung cancer death rates between men and women and between African Americans and whites and analyzed temporal trends in lung cancer death rates among never smokers across the two studies by using directly age-standardized rates as well as Poisson and Cox proportional hazards regression analyses. All statistical tests were two-sided. RESULTS: The age-standardized lung cancer death rates among never-smoking men and women in CPS-II were 17.1 and 14.7 per 100,000 person-years, respectively. Men who had never smoked had higher age-standardized lung cancer death rates than women in both studies (CPS-I: hazard ratio [HR] = 1.52, 95% confidence interval [CI] = 1.28 to 1.79; CPS-II: HR = 1.21, 95% CI = 1.09 to 1.36). The rate was higher among African American women than white women in CPS-II (HR = 1.43, CI = 1.11 to 1.85). A small temporal increase (CPS-II versus CPS-I) in lung cancer mortality was seen for white women (HR = 1.25, CI = 1.12 to 1.41) and African American women (HR = 1.22, CI = 0.64 to 2.33), but not for white men (HR = 0.89, CI = 0.74 to 1.08). Among white and African American women combined, the temporal increase was statistically significant only among those aged 70-84 years (P < .001). CONCLUSIONS: Contrary to clinical perception, the lung cancer death rate is not higher in female than in male never smokers and shows little evidence of having increased over time in the absence of smoking. Factors that affect the interpretation of lung cancer trends are discussed. Our novel finding that lung cancer mortality is higher among African American than white women never smokers should be confirmed in other studies. PMID- 16705124 TI - Statin use and breast cancer: prospective results from the Women's Health Initiative. AB - BACKGROUND: Despite experimental observations suggesting that 3-hydroxy-3 methylglutaryl coenzyme A inhibitors (statins) have antitumor activity, clinical studies have reached mixed conclusions about the relationship between statin use and breast cancer risk. METHODS: To investigate associations between potency, duration of use, and type of statin used and risk of invasive breast cancer, we examined data for 156,351 postmenopausal women who were enrolled in the Women's Health Initiative. Information was collected on breast cancer risk factors and on the use of statins and other lipid-lowering drugs. Cox proportional hazards regression was used to calculate hazard ratios (HRs) with 95% confidence intervals (CIs). Statistical tests were two-sided. RESULTS: Over an average follow-up of 6.7 years, 4383 invasive breast cancers were confirmed by medical record and pathology report review. Statins were used by 11,710 (7.5%) of the cohort. Breast cancer incidence was 4.09 per 1000 person-years (PY) among statin users and 4.28 per 1000 PY among nonusers. In multivariable models, the hazard ratio of breast cancer among users of any statin, compared with nonusers, was 0.91 (95% CI = 0.80 to 1.05, P = .20). There was no trend in risk by duration of statin use, with HR = 0.80 (95% CI = 0.63 to 1.03) for < 1 year of use, HR = 0.99 (95% CI = 0.80 to 1.23) for 1- < 3 years of use, and HR = 0.94 (95% CI = 0.75 to 1.18) for > or = 3 years of use. Hydrophobic statins (i.e., simvastatin, lovastatin, and fluvastatin) were used by 8106 women, and their use was associated with an 18% lower breast cancer incidence (HR = 0.82, 95% CI = 0.70 to 0.97, P = .02). Use of other statins (i.e., pravastatin and atorvastatin) or nonstatin lipid-lowering agents was not associated with breast cancer incidence. CONCLUSIONS: Overall statin use was not associated with invasive breast cancer incidence. Our finding that use of hydrophobic statins may be associated with lower breast cancer incidence suggests possible within-class differences that warrant further evaluation. PMID- 16705125 TI - Recombinant human erythropoietins and cancer patients: updated meta-analysis of 57 studies including 9353 patients. AB - This is an updated systematic review of 57 trials and 9353 cancer patients from articles, abstracts, and reports published between January 1, 1985, and April 30, 2005, on the effects of epoetin alfa and beta (i.e., epoetin) and darbepoetin alfa (i.e., darbepoetin). We included randomized controlled trials comparing epoetin or darbepoetin plus red blood cell transfusion with red blood cell transfusion alone for prophylaxis or treatment of anemia in cancer patients with or without concurrent antineoplastic therapy. The Cochrane Library, MEDLINE, EMBASE, and conference proceedings were searched. Effect estimates and 95% confidence intervals (CIs) were calculated with fixed-effects models. Treatment with epoetin or darbepoetin statistically significantly reduced the risk for red blood cell transfusions (relative risk [RR] = 0.64, 95% CI = 0.60 to 0.68; 42 trials and 6510 patients) and improved hematologic response (RR = 3.43, 95% CI = 3.07 to 3.84; 22 trials and 4307 patients). Treatment with epoetin or darbepoetin increased the risk of thrombo-embolic events (RR = 1.67, 95% CI = 1.35 to 2.06; 35 trials and 6769 patients). Uncertainties remain as to whether and how epoetin or darbepoetin affects overall survival (hazard ratio = 1.08, 95% CI = 0.99 to 1.18; 42 trials and 8167 patients). Caution is advised when using epoetin or darbepoetin in combination with thrombogenic chemotherapeutic agents or for cancer patients who are at high risk for thrombo-embolic events. PMID- 16705126 TI - Preoperative nomogram predicting the 10-year probability of prostate cancer recurrence after radical prostatectomy. AB - An existing preoperative nomogram predicts the probability of prostate cancer recurrence, defined by prostate-specific antigen (PSA), at 5 years after radical prostatectomy based on clinical stage, serum PSA, and biopsy Gleason grade. In an updated and enhanced nomogram, we have extended the predictions to 10 years, added the prognostic information of systematic biopsy results, and enabled the predictions to be adjusted for the year of surgery. Cox regression analysis was used to model the clinical information for 1978 patients treated by two high volume surgeons from our institution. The nomogram was externally validated on an independent cohort of 1545 patients with a concordance index of 0.79 and was well calibrated with respect to observed outcome. The inclusion of the number of positive and negative biopsy cores enhanced the predictive accuracy of the model. Thus, a new preoperative nomogram provides robust predictions of prostate cancer recurrence up to 10 years after radical prostatectomy. PMID- 16705127 TI - Re: Effect of gamma-linolenic acid on the transcriptional activity of the Her 2/neu (erbB-2) oncogene. PMID- 16705129 TI - Viral modulators of cullin RING ubiquitin ligases: culling the host defense. AB - Cullin RING ubiquitin ligases (CRULs) are found in all eukaryotes and play an essential role in targeting proteins for ubiquitin-mediated destruction, thus regulating a plethora of cellular processes. Viruses manipulate CRULs by redirecting this destruction machinery to eliminate unwanted host cell proteins, thus allowing viruses to slip past host immune barriers. Depending on the host organism, virus-modified CRULs can perform an amazing range of tasks, including the elimination of crucial signal transduction molecules in the human interferon pathway and suppression of virus-induced gene silencing in plants. This Perspective summarizes recent advances in our understanding of how viral proteins manipulate the function of CRULs. PMID- 16705130 TI - Notch and Wnt signaling: mimicry and manipulation by gamma herpesviruses. AB - A small number of fundamental cell signaling pathways are key to the regulation of proliferation and differentiation responses during normal development. Two of these pathways, the Notch and Wnt pathways, have proven to be attractive targets for virus interaction and manipulation. In general, viral gene expression and replication are intimately linked to the differentiation state of the infected cell and, in the case of the gamma herpesviruses, establishment of a lifelong persistent infection in the host is also dependent on the proliferative expansion of an infected B cell population. This review examines the ways in which the gamma herpesviruses Epstein-Barr virus (EBV) and Kaposi's sarcoma-associated herpesvirus (KSHV) have exploited the Notch and Wnt pathways to advance their own life cycles. The virus-pathway interactions are compared with the mechanisms and outcome of cellular Notch and Wnt signaling. PMID- 16705131 TI - Signaling during pathogen infection. AB - Over the millennia, pathogens have coevolved with their hosts and acquired the ability to intercept, disrupt, mimic, and usurp numerous signaling pathways of those hosts. The study of host/pathogen interactions thus not only teaches us about the intricate biology of these parasitic invaders but also provides interesting insights into basic cellular processes both at the level of the individual cell and more globally throughout the organism. Host/pathogen relationships also provide insights into the evolutionary forces that shape biological diversity. Here we review a few recent examples of how viruses, bacteria, and parasites manipulate tyrosine kinase-mediated and Rho guanosine triphosphatase-mediated signaling pathways of their hosts to achieve efficient entry, replication, and exit during their infectious cycles. PMID- 16705132 TI - Depletion of serotonin selectively impairs short-term memory without affecting long-term memory in odor learning in the terrestrial slug Limax valentianus. AB - The terrestrial slug Limax is able to acquire short-term and long-term memories during aversive odor-taste associative learning. We investigated the effect of the selective serotonergic neurotoxin 5,7-dihydroxytryptamine (5,7-DHT) on memory. Behavioral studies indicated that 5,7-DHT impaired short-term memory but not long-term memory. HPLC (high-performance liquid chromatography) analysis revealed that 5,7-DHT significantly reduced serotonin content in the central nervous system. The present study suggests that acquisition, retention, and/or retrieval of short-term memory involves serotonin, and neither acquisition nor retrieval of long-term memory requires serotonin at a level as high as that required for short-term memory. PMID- 16705133 TI - The characteristics of LTP induced in hippocampal slices are dependent on slice recovery conditions. AB - In area CA1 of hippocampal slices which are allowed to recover from slicing "in interface" and where recordings are carried out in interface, a single 1-sec train of 100-Hz stimulation triggers a short-lasting long-term potentiation (S LTP), which lasts 1-2 h, whereas multiple 1-sec trains induce a long-lasting LTP (L-LTP), which lasts several hours. Moreover, the threshold and the features of these LTP depend on the history of the neurons, a phenomenon known as metaplasticity. Here, where all recordings were performed in interface, we found that allowing the slices to recover "in submersion" had dramatic metaplastic effects. In these conditions, a single 1-sec train at 100 Hz induced an L-LTP which lasted at least 4 h and was dependent on protein synthesis. Interestingly, this type of metaplasticity was observed when the concentration of Mg(++) used was 1.0 mM but not when it was 1.3 mM. The LTP induced by four 1-sec trains at 100 Hz was similar whatever the incubation method. However, the signaling cascades recruited to achieve that pattern were different. In the interface interface paradigm (recovery and recording both in interface) the four-train induced LTP recruited the PKA signaling pathway but not that of the p42/44MAPK. On the contrary, in the submersion-interface paradigm the four-train induced LTP recruited the p42/44MAPK signaling pathway but not that of the PKA. To our knowledge this is the first example of metaplasticity involving the recruitment of signaling cascades in LTP. PMID- 16705134 TI - Place learning in the Morris water task: making the memory stick. AB - Although the Morris water task has been used in hundreds of studies of place learning, there have been no systematic studies of retention of the place memory. We report that retention, as measured by selective search behavior on a probe trial, is excellent when the retention interval is short (5-10 min). However, performance rapidly deteriorates, so that by approximately 4 h the search is no longer selective. Additional experiments revealed that selective search at longer retention intervals was improved by inserting gaps between blocks of training trials, but this effect was a non-monotonic function of the interval separating trial blocks. Our experiments also revealed that the location of the first block of trials (Room A or Room B) was irrelevant to long-term retention. A memory modulation theoretical framework may provide a useful way to understand these findings. PMID- 16705135 TI - Repeated acquisitions and extinctions in classical conditioning of the rabbit nictitating membrane response. AB - The rabbit nictitating membrane (NM) response underwent successive stages of acquisition and extinction training in both delay (Experiment 1) and trace (Experiment 2) classical conditioning. In both cases, successive acquisitions became progressively faster, although the largest, most reliable acceleration occurred between the first and second acquisition. Successive extinctions were similar in rate. The results challenge contextual control theories of extinction but are consistent with attentional and layered-network models. The results are discussed with respect to their implications for the interaction between cerebellar and forebrain pathways for eyeblink conditioning. PMID- 16705136 TI - Post-training intrahippocampal injection of synthetic poly-alpha-2,8-sialic acid neural cell adhesion molecule mimetic peptide improves spatial long-term performance in mice. AB - Several data have shown that the neural cell adhesion molecule (NCAM) is necessary for long-term memory formation and might play a role in the structural reorganization of synapses. The NCAM, encoded by a single gene, is represented by several isoforms that differ with regard to their content of alpha-2,8-linked sialic acid residues (PSA) on their extracellular domain. The carbohydrate PSA is known to promote plasticity, and PSA-NCAM isoforms remain expressed in the CA3 region of the adult hippocampus. In the present study, we investigated the effect on spatial memory consolidation of a PSA gain of function by injecting a PSA mimetic peptide (termed pr2) into the dorsal hippocampus. Mice were subjected to massed training in the spatial version of the water maze. Five hours after the last training session, experimental mice received an injection of pr2, whereas control mice received PBS or reverse peptide injections in the hippocampal CA3 region. Memory retention was tested at different time intervals: 24 h, 1 wk, and 4 wk. The results showed that the post-training infusion of pr2 peptide significantly increases spatial performance whenever it was assessed after the training phase. By contrast, administration of the control reverse peptide did not affect retention performance. These findings provide evidence that (1) PSA NCAM is involved in memory consolidation processes in the CA3 hippocampal region, and (2) PSA mimetic peptides can facilitate the formation of long-term spatial memory when injected during the memory consolidation phase. PMID- 16705137 TI - Effects of intra-amygdala infusion of CB1 receptor agonists on the reconsolidation of fear-potentiated startle. AB - The cannabinoid CB1 receptor has been shown to be critically involved in the extinction of fear memory. Systemic injection of a CB1 receptor antagonist prior to extinction training blocked extinction. Conversely, administration of the cannabinoid uptake inhibitor AM404 facilitated extinction in a dose-dependent manner. Here we show that bilateral infusion of CB1 receptor agonists into the amygdala after memory reactivation blocked reconsolidation of fear memory measured with fear-potentiated startle. The effect was dose-dependent and could be blocked by AM251, a specific CB1 receptor antagonist. In contrast, the effect of CB1 agonists on reconsolidation was no longer seen if memory reactivation was omitted. Concomitant with block of reconsolidation, CB1 agonist-treated animals did not exhibit shock-induced reinstatement or spontaneous recovery of fear. The absence of recovery was not attributable to permanent damage to the amygdala in WIN-treated rats, nor did the effect result from alteration of baseline startle or shock reactivity. These results suggest that CB1 agonists could impair fear memory via blocking reconsolidation. PMID- 16705138 TI - Dishabituation in Aplysia can involve either reversal of habituation or superimposed sensitization. AB - Dishabituation has been thought to be due either to reversal of the process of habituation or to a second process equivalent to sensitization superimposed on habituation. One way to address this question is by testing whether dishabituation and sensitization can be dissociated. Previous studies using this approach in Aplysia have come to different conclusions about the nature of dishabituation, perhaps because those studies differed in many respects, including (1) whether they also observed transient behavioral inhibition, and (2) whether they used test stimuli that activated the LE siphon sensory neurons or as yet unidentified sensory neurons. To attempt to resolve the apparent contradictions between the previous studies, we have explored the importance of these two factors by performing a parametric study of dishabituation and sensitization of gill withdrawal in a simplified preparation that does not exhibit transient behavioral inhibition, using two different test stimuli that are known to activate the LE (Touch) or unidentified (Not Touch) sensory neurons. We find that dishabituation and sensitization in this preparation have similar time courses and generally similar functions of shock intensity. However, under one condition, with the Not Touch stimulus 2.5 min after the shock, dishabituation has a reverse effect of shock intensity. Additional analyses suggest that dishabituation with the Not Touch stimulus 2.5 min after the shock is due to reversal of habituation, whereas 12.5 min after the shock, dishabituation is due to superimposed sensitization. These results thus suggest that dishabituation may involve either process in the same preparation, and begin to define the conditions that favor one or the other. PMID- 16705139 TI - NT-3 facilitates hippocampal plasticity and learning and memory by regulating neurogenesis. AB - In the adult brain, the expression of NT-3 is largely confined to the hippocampal dentate gyrus (DG), an area exhibiting significant neurogenesis. Using a conditional mutant line in which the NT-3 gene is deleted in the brain, we investigated the role of NT-3 in adult neurogenesis, hippocampal plasticity, and memory. Bromodeoxyuridine (BrdU)-labeling experiments demonstrated that differentiation, rather than proliferation, of the neuronal precursor cells (NPCs) was significantly impaired in DG lacking NT-3. Triple labeling for BrdU, the neuronal marker NeuN, and the glial marker GFAP indicated that NT-3 affects the number of newly differentiated neurons, but not glia, in DG. Field recordings revealed a selective impairment in long-term potentiation (LTP) in the lateral, but not medial perforant path-granule neuron synapses. In parallel, the NT-3 mutant mice exhibited deficits in spatial memory tasks. In addition to identifying a novel role for NT-3 in adult NPC differentiation in vivo, our study provides a potential link between neurogenesis, dentate LTP, and spatial memory. PMID- 16705140 TI - Foreground contextual fear memory consolidation requires two independent phases of hippocampal ERK/CREB activation. AB - Fear conditioning is a popular model for investigating physiological and cellular mechanisms of memory formation. In this paradigm, a footshock is either systematically associated to a tone (paired conditioning) or is pseudorandomly distributed (unpaired conditioning). In the former procedure, the tone/shock association is acquired, whereas in the latter procedure, the context/shock association will prevail. Animals with chronically implanted recording electrodes show enhanced amplitude of the extracellularly recorded field EPSP in CA1 pyramidal cells for up to 24 h after unpaired, but not paired, fear conditioning. This is paralleled by a differential activation of the ERK/CREB pathway in CA1, which is monophasic in paired conditioning (0-15 min post-conditioning), but biphasic (0-1 h and 9-12 h post-conditioning) in unpaired conditioning as revealed by immunocytochemistry and Western blotting. Intrahippocampal injection of the MEK inhibitor U0126 prior to each phase prevents the activation of both ERK1/2 and CREB after unpaired conditioning. Block of any activation phase leads to memory impairment. We finally reveal that the biphasic activation of ERK/CREB activity is independently regulated, yet both phases are critically required for the consolidation of long-term memories following unpaired fear conditioning. These data provide compelling evidence that CA1 serves different forms of memory by expressing differential cellular mechanisms that are dependent on the training regime. PMID- 16705141 TI - Do rats use shape to solve "shape discriminations"? AB - Visual discrimination tasks are increasingly used to explore the neurobiology of vision in rodents, but it remains unclear how the animals solve these tasks: Do they process shapes holistically, or by using low-level features such as luminance and angle acuity? In the present study we found that when discriminating triangles from squares, rats did not use shape but instead relied on local luminance differences in the lower hemifield. A second experiment prevented this strategy by using stimuli-squares and rectangles-that varied in size and location, and for which the only constant predictor of reward was aspect ratio (ratio of height to width: a simple descriptor of "shape"). Rats eventually learned to use aspect ratio but only when no other discriminand was available, and performance remained very poor even at asymptote. These results suggest that although rats can process both dimensions simultaneously, they do not naturally solve shape discrimination tasks this way. This may reflect either a failure to visually process global shape information or a failure to discover shape as the discriminative stimulus in a simultaneous discrimination. Either way, our results suggest that simultaneous shape discrimination is not a good task for studies of visual perception in rodents. PMID- 16705142 TI - Advances in the treatment of prolactinomas. AB - Prolactinomas account for approximately 40% of all pituitary adenomas and are an important cause of hypogonadism and infertility. The ultimate goal of therapy for prolactinomas is restoration or achievement of eugonadism through the normalization of hyperprolactinemia and control of tumor mass. Medical therapy with dopamine agonists is highly effective in the majority of cases and represents the mainstay of therapy. Recent data indicating successful withdrawal of these agents in a subset of patients challenge the previously held concept that medical therapy is a lifelong requirement. Complicated situations, such as those encountered in resistance to dopamine agonists, pregnancy, and giant or malignant prolactinomas, may require multimodal therapy involving surgery, radiotherapy, or both. Progress in elucidating the mechanisms underlying the pathogenesis of prolactinomas may enable future development of novel molecular therapies for treatment-resistant cases. This review provides a critical analysis of the efficacy and safety of the various modes of therapy available for the treatment of patients with prolactinomas with an emphasis on challenging situations, a discussion of the data regarding withdrawal of medical therapy, and a foreshadowing of novel approaches to therapy that may become available in the future. PMID- 16705143 TI - Role of a tyrosine kinase in the CO2-induced stimulation of HCO3- reabsorption by rabbit S2 proximal tubules. AB - A previous study demonstrated that proximal tubule cells regulate HCO(3)(-) reabsorption by sensing acute changes in basolateral CO(2) concentration, suggesting that there is some sort of CO(2) sensor at or near the basolateral membrane (Zhou Y, Zhao J, Bouyer P, and Boron WF Proc Natl Acad Sci USA 102: 3875 3880, 2005). Here, we hypothesized that an early element in the CO(2) signal transduction cascade might be either a receptor tyrosine kinase (RTK) or a receptor-associated (or soluble) tyrosine kinase (sTK). In our experiments, we found, first, that basolateral 17.5 microM genistein, a broad-spectrum tyrosine kinase inhibitor, virtually eliminates the CO(2) sensitivity of HCO(3)(-) absorption rate (J(HCO(3))). Second, we found that neither basolateral 250 nM nor basolateral 2 microM PP2, a high-affinity inhibitor for the Src family that also inhibits the Bcr-Abl sTK as well as the Kit RTK, reduces the CO(2)-stimulated increase in J(HCO(3)). Third, we found that either basolateral 35 nM PD168393, a high-affinity inhibitor of RTKs in the erbB (i.e., EGF receptor) family, or basolateral 10 nM BPIQ-I, which blocks erbB RTKs by competing with ATP, eliminates the CO(2) sensitivity. In conclusion, the transduction of the CO(2) signal requires activation of a tyrosine kinase, perhaps an erbB. The possibilities include the following: 1) a TK is simply permissive for the effect of CO(2) on J(HCO(3)); 2) a CO(2) receptor activates an sTK, which would then raise J(HCO(3)); 3) a CO(2) receptor transactivates an RTK; and 4) the CO(2) receptor could itself be an RTK. PMID- 16705144 TI - Spontaneous activation of the NF-kappaB signaling pathway in isolated normal glomeruli. AB - In this report, we describe that NF-kappaB is spontaneously activated in isolated, normal glomeruli. Ex vivo incubation of isolated rat glomeruli triggered expression of a NF-kappaB-dependent gene, monocyte chemoattractant protein-1 (MCP-1), in parallel with downregulation of IkappaBalpha and IkappaBbeta proteins and activation of the p65 NF-kappaB subunit. The induction of MCP-1 was also observed in mesangial cells coincubated with isolated glomeruli or exposed to media conditioned by isolated glomeruli (GCM), which was abrogated by inhibition of NF-kappaB. The activation of NF-kappaB by glomerulus-derived factors was confirmed using reporter mesangial cells that produce secreted alkaline phosphatase (SEAP) under the control of the kappaB enhancer element. When the reporter cells were adoptively transferred into normal glomeruli, expression of SEAP mRNA and activity of SEAP were also upregulated in the explanted glomeruli. The molecular weight of factors responsible for activation of NF-kappaB was >50 kDa, and TNF-alpha was identified as one of glomerulus derived activators. To examine upstream events involved, we focused on MAP kinases that are spontaneously activated in explanted glomeruli. Selective suppression of ERK or p38 MAP kinase significantly attenuated activation of NF kappaB in mesangial cells triggered by coculture with isolated glomeruli. Interestingly, the suppressive effects by MAP kinase inhibitors were not observed in mesangial cells treated with GCM. These data suggested that NF-kappaB was spontaneously activated in explanted glomeruli via autocrine/paracrine factors including TNF-alpha and that the production of NF-kappaB activators by glomeruli was, at least in part, through MAP kinase pathways. PMID- 16705145 TI - Nitric oxide stimulates COX-2 expression in cultured collecting duct cells through MAP kinases and superoxide but not cGMP. AB - Collecting ducts are a major site of renal production and action of both prostaglandins and nitric oxide. Experiments were undertaken to examine whether nitric oxide regulates cyclooxygenase (COX)-2 expression and PGE(2) release in cultured collecting duct cells. In mIMCD-K2 cells, sodium nitroprusside (SNP) in the 50- to 800-microM range induced a marked dose- and time-dependent increase in COX-2 protein levels, determined by immunoblotting, and the induction was detectable at 4 h. This was preceded by induction of COX-2 mRNA as determined by real-time-RT-PCR. The COX-2 induction was accompanied by a significant rise in PGE(2) release as determined by enzyme immunoassay. S-nitroso-N acetylpenicillamine (SNAP) had a similar stimulatory effect on COX-2 expression and PGE(2) release. 8-bromo-cGMP (200 microM) had no effect on COX-2 expression. The SNP-stimulated COX-2 expression was not affected by the guanylyl cyclase inhibitor methylene blue or the protein kinase G inhibitor KT-5823 (2.0 microM). In contrast, the SNP-stimulated COX-2 expression was significantly reduced by either the Erk1/2 inhibitor PD-98059 or the P38 inhibitor SB-203580 and was abolished by combination of the two kinase inhibitors. The stimulation was also significantly blocked by the SOD mimetic tempol. Thus we conclude that NO stimulates COX-2 expression in collecting duct cells through mechanisms involving MAP kinase and superoxide, but not cGMP. PMID- 16705146 TI - Characterization of diabetic nephropathy in a transgenic model of hypoinsulinemic diabetes. AB - Genetic mouse models provide a unique opportunity to investigate gene function in the natural course of the disease. Although diabetic nephropathy (DN) in models of type II diabetes has been well characterized, diabetic renal disease in hypoinsulinemic diabetic mice is still incompletely understood. Here, we characterized renal changes in the pdx1(PB)-HNF6 transgenic mouse that exhibits beta-cell dysfunction and nonobese hypoinsulinemic diabetes. Male transgenic mice developed hyperglycemia by the age of 7 wk and survived for over 1 yr without insulin treatment. Diabetes ensued earlier and progressed more severely in the HNF6 males than the females. The HNF6 males exhibited albuminuria as early as 10 wk of age, and the urinary albumin excretion increased with age, exceeding 150 microg/24 h at 11 mo of age. Diabetic males developed renal hypertrophy after 7 wk of age, whereas glomerular hyperfiltration was not observed in the mice. Hypertension and hyperlipidemia were not observed in the diabetic mice. Histological analysis of the HNF6 kidneys displayed diabetic glomerular changes, including glomerular enlargement, diffuse mesangial proliferation and matrix expansion, thickened glomerular basement membrane, and arteriolar hyalinosis. Mesangial matrix accumulation increased with age, resulting in nodular lesions by 44 wk of age. Immunohistochemistry showed accumulation of type IV collagen and TGF-beta1 in the mesangial area. No significant immune complex deposition was observed in the HNF6 glomeruli. Thus the HNF6 mouse exhibits diabetic renal changes that parallel the early phase of human DN. The model should facilitate studies of genetic and environmental factors that may affect DN in hypoinsulinemic diabetes. PMID- 16705148 TI - Thiamin uptake by the human-derived renal epithelial (HEK-293) cells: cellular and molecular mechanisms. AB - Thiamin (vitamin B(1)) is essential for normal cellular functions. The kidneys play a critical role in regulating body thiamin homeostasis, by salvaging the vitamin via reabsorption from the glomerular filtrate, but little is known about the mechanism(s) and regulation of thiamin transport in the human renal epithelia at cellular and molecular levels. Using the human-derived renal epithelial HEK 293 cells as a model, we have addressed these issues. Our results showed [(3)H]thiamin uptake to be 1) temperature and energy dependent but Na(+) independent, 2) pH dependent with higher uptake at alkaline/neutral buffer pH compared with acidic pH, 3) saturable as a function of concentration over the nanomolar (apparent K(m) = 70.0 +/- 18.4 nM) and micromolar (apparent K(m) = 2.66 +/- 0.18 microM) ranges, 4) cis-inhibited by unlabeled thiamin and its structural analogs but not by unrelated organic cations, 5) trans-stimulated by unlabeled thiamin, and 6) competitively inhibited by amiloride with an apparent K(i) of 0.6 mM. Using a gene-specific small-interference RNAs (siRNAs) approach, human thiamin transporters 1 and 2 (hTHTR-1 and hTHTR-2) were both found to be expressed and contributed toward total carrier-mediated thiamin uptake. Maintaining the cells in thiamin-deficient medium led to a significant (P < 0.01) and specific upregulation in [(3)H]thiamin uptake, which was associated with an increase in hTHTR-1 and hTHTR-2 protein and mRNA levels as well as promoter activities. Uptake of thiamin by HEK-293 cells also appeared to be under the regulation of an intracellular Ca(2+)/calmodulin-mediated pathway. These studies demonstrate for the first time that thiamin uptake by HEK-293 cells is mediated via a specific pH-dependent process, which involves both the hTHTR-1 and hTHTR-2. In addition, the uptake process appears to be under the regulation of an intracellular Ca(2+)/CaM-mediated pathway and also adaptively upregulated in thiamin deficiency via transcriptional regulatory mechanism(s) that involves both the hTHTR-1 and hTHTR-2. PMID- 16705147 TI - Activation of ERK1/2 pathway mediates oxidant-induced decreases in mitochondrial function in renal cells. AB - Previously, we showed that oxidant exposure in renal proximal tubular cells (RPTC) induces mitochondrial dysfunction mediated by PKC-epsilon. This study examined the role of ERK1/2 in mitochondrial dysfunction induced by oxidant injury and whether PKC-epsilon mediates its effects on mitochondrial function through the Raf-MEK1/2-ERK1/2 pathway. Sublethal injury produced by tert butylhydroperoxide (TBHP) resulted in three- to fivefold increase in phosphorylation of ERK1/2 and p38 but not JNK. This was followed by decreases in basal and uncoupled respirations (41%), state 3 respiration and ATP production coupled to complex I (46%), and complex I activity (42%). Oxidant exposure decreased aconitase activity 30% but not pyruvate, alpha-ketoglutarate, and malate dehydrogenase activities. Inhibition of ERK1/2 restored basal and state 3 respirations, DeltaPsi(m), ATP production, and complex I activity but not aconitase activity. In contrast, activation of ERK1/2 by expression of constitutively active MEK1 suppressed basal, uncoupled, and state 3 respirations in noninjured RPTC to the levels observed in TBHP-injured RPTC. MEK1/2 inhibition did not change Akt or p38 phosphorylation, demonstrating that the protective effect of MEK1/2 inhibitor was not due to activation of Akt or inhibition of p38 pathway. Inhibition of PKC-epsilon did not block TBHP-induced ERK1/2 phosphorylation in whole RPTC or in mitochondria. We conclude that 1) oxidant induced activation of ERK1/2 but not p38 or JNK reduces mitochondrial respiration and ATP production by decreasing complex I activity and substrate oxidation through complex I, 2) citric acid cycle dehydrogenases are not under control of the ERK1/2 pathway in oxidant-injured RPTC, 3) the protective effects of ERK1/2 inhibition are not due to activation of Akt, and 4) ERK1/2 and PKC-epsilon mediate oxidant-induced mitochondrial dysfunction through independent pathways. PMID- 16705149 TI - Selective upregulation of the ADP-ribosyl cyclases CD38 and CD157 by TNF but not by RANK-L reveals differences in downstream signaling. AB - In macrophages and osteoclast precursors, the cytokines TNF and RANK-L induce similar downstream pathways and share some of the same adaptor molecules. However, despite these similarities, no defined signaling schematic has emerged to show how each cytokine favors particular pathways. In this report, we investigate whether TNF and RANK-L differentially regulate ADP-ribosyl cyclases enzymes that are unique in being crucial for immunological function yet detrimental to osteoclastogenesis. TNF but not RANK-L led to the sustained upregulation of both CD38 and CD157 as demonstrated by real-time PCR and flow cytometry. Further investigation demonstrated that this upregulation was a result of continuous, direct TNF signaling and involved JNK, and more critically PKC and NF-kappaB. Using this approach allowed us to highlight the relative importance of the PKC, NF-kappaB, and JNK pathways in actualizing proper outcomes of TNF signaling. Albeit speculative, we believe that differences between TNF- and RANK l-induced activation of downstream signaling pathways, in particular PKC, are crucial for determining whether progenitor cells become geared for immunity or bone resorption. PMID- 16705150 TI - Regulation of ecto-5'-nucleotidase by NaCl and nitric oxide: potential roles in tubuloglomerular feedback and adaptation. AB - The tubuloglomerular feedback (TGF) system serves to establish an appropriate balance between tubular reabsorption and glomerular filtration rate (GFR). High salt at the macula densa activates TGF to decrease GFR. Effector molecules for the TGF signal include ATP and adenosine. Over time, the GFR will adapt by increasing even if a high salt concentration persists. A potential modulator of this TGF adaptation is nitric oxide synthase-1-derived nitric oxide (NO). In isolated glomerular preparations, we developed a system for evaluating the effects of changing dietary salt on ecto-5'-nucleotidase (ecto-5'-NT) activity, the final enzyme in the conversion of ATP to adenosine. We found observable ecto 5'-NT activity in isolated glomeruli and that this activity can be regulated by dietary salt, with high salt increasing activity. Conversely, NO decreases ecto 5'-NT activity in glomerular preparations. Moreover, NO inhibition of ecto-5'-NT activity is suppressed in the presence of dithiothreitol, suggesting nitrosylation as a reversible, oxidative stress-sensitive mechanism. The salt induced activation of ecto-5'-NT correlates with high salt resetting of TGF. NO inhibition of enzymatic activity could be part of the adaptive phase. PMID- 16705151 TI - Age-dependent alterations in Ca2+ homeostasis: role of TRPV5 and TRPV6. AB - Aging is associated with alterations in Ca2+ homeostasis, which predisposes elder people to hyperparathyroidism and osteoporosis. Intestinal Ca2+ absorption decreases with aging and, in particular, active transport of Ca2+ by the duodenum. In addition, there are age-related changes in renal Ca2+ handling. To examine age-related changes in expression of the renal and intestinal epithelial Ca2+ channels, control (TRPV5+/+) and TRPV5 knockout (TRPV5-/-) mice aged 10, 30, and 52 wk were studied. Aging of TRPV5(+/+) mice resulted in a tendency toward increased renal Ca2+ excretion and significantly decreased intestinal Ca2+ absorption, which was accompanied by reduced expression of TRPV5 and TRPV6, respectively, despite increased serum 1,25(OH)2D3 levels. Similarly, in TRPV5-/- mice the existing renal Ca2+ loss was more pronounced in elder animals, whereas the compensatory intestinal Ca2+ absorption and TRPV6 expression declined with aging. In both mice strains, aging resulted in a resistance to 1,25(OH)2D3 and diminished renal vitamin D receptor mRNA levels, whereas serum Ca2+ levels remained constant. Furthermore, 52-wk-old TRPV5-/- mice showed severe hyperparathyroidism, whereas PTH levels in elder TRPV5+/+ mice remained normal. In 52-wk-old TRPV5-/- mice, serum osteocalcin levels were increased in accordance with the elevated PTH levels, suggesting an increased bone turnover in these mice. In conclusion, downregulation of TRPV5 and TRPV6 is likely involved in the impaired Ca2+ (re)absorption during aging. Moreover, TRPV5-/- mice likely develop age-related hyperparathyroidism and osteoporotic characteristics before TRPV5+/+ mice, demonstrating the importance of the epithelial Ca2+ channels in Ca2+ homeostasis. PMID- 16705152 TI - Adenoviral expression of NHERF-1 in NHERF-1 null mouse renal proximal tubule cells restores Npt2a regulation by low phosphate media and parathyroid hormone. AB - Sodium-dependent phosphate transport in NHERF-1(-/-) proximal tubule cells does not increase when grown in a low phosphate media and is resistant to the normal inhibitory effects of parathyroid hormone (PTH). The current experiments employ adenovirus-mediated gene transfer in primary cultures of mouse proximal tubule cells from NHERF-1 null mice to explore the specific role of NHERF-1 on regulated Npt2a trafficking and sodium-dependent phosphate transport. NHERF-1 null cells have decreased sodium-dependent phosphate transport compared with wild-type cells. Infection of NHERF-1 null cells with adenovirus-GFP-NHERF-1 increased phosphate transport and plasma membrane abundance of Npt2a. Adenovirus-GFP-NHERF 1 infected NHERF-1 null proximal tubule cells but not cells infected with adenovirus-GFP demonstrated increased phosphate transport and Npt2a abundance in the plasma membrane when grown in low phosphate (0.1 mM) compared with high phosphate media (1.9 mM). PTH inhibited phosphate transport and decreased Npt2a abundance in the plasma membrane of adenovirus-GFP-NHERF-1-infected NHERF-1 null proximal tubule cells but not cells infected with adenovirus-GFP. Interestingly, phosphate transport is inhibited by activation of protein kinase A and protein kinase C in wild-type proximal tubule cells but not in NHERF-1(-/-) cells. Together, these results highlight the requirement for NHERF-1 for physiological control of Npt2a trafficking and suggest that the Npt2a/NHERF-1 complex represents a unique PTH-responsive pool of Npt2a in renal microvilli. PMID- 16705153 TI - Endothelin and nitric oxide mediate adaptation of the cortical collecting duct to metabolic acidosis. AB - Endothelin (ET) and nitric oxide (NO) modulate ion transport in the kidney. In this study, we defined the function of ET receptor subtypes and the NO guanylate cyclase signaling pathway in mediating the adaptation of the rabbit cortical collecting duct (CCD) to metabolic acidosis. CCDs were perfused in vitro and incubated for 3 h at pH 6.8, and bicarbonate transport or cell pH was measured before and after acid incubation. Luminal chloride was reversibly removed to isolate H(+) and HCO(3)(-) secretory fluxes and to raise the pH of beta intercalated cells. Acid incubation caused reversal of polarity of net HCO(3)(-) transport from secretion to absorption, comprised of a 40% increase in H(+) secretion and a 75% decrease in HCO(3)(-) secretion. The ET(B) receptor antagonist BQ-788, as well as the NO synthase inhibitor, N(G)-nitro-l-arginine methyl ester (l-NAME), attenuated the adaptive decrease in HCO(3)(-) secretion by 40%, but only BQ-788 inhibited the adaptive increase in H(+) secretion. There was no effect of inactive d-NAME or the ET(A) receptor antagonist BQ-123. Both BQ-788 and l-NAME inhibited the acid-induced inactivation (endocytosis) of the apical Cl(-)/HCO(3)(-) exchanger. The guanylate cyclase inhibitor LY-83583 and cGMP dependent protein kinase inhibitor KT-5823 affected HCO(3)(-) transport similarly to l-NAME. These data indicate that signaling via the ET(B) receptor regulates the adaptation of the CCD to metabolic acidosis and that the NO guanylate cyclase component of ET(B) receptor signaling mediates downregulation of Cl(-)/HCO(3)(-) exchange and HCO(3)(-) secretion. PMID- 16705154 TI - Requirement of ELC1 for RNA polymerase II polyubiquitylation and degradation in response to DNA damage in Saccharomyces cerevisiae. AB - Treatment of Saccharomyces cerevisiae and human cells with DNA-damaging agents such as UV light or 4-nitroquinoline-1-oxide induces polyubiquitylation of the largest RNA polymerase II (Pol II) subunit, Rpb1, which results in rapid Pol II degradation by the proteasome. Here we identify a novel role for the yeast Elc1 protein in mediating Pol II polyubiquitylation and degradation in DNA-damaged yeast cells and propose the involvement of a ubiquitin ligase, of which Elc1 is a component, in this process. In addition, we present genetic evidence for a possible involvement of Elc1 in Rad7-Rad16-dependent nucleotide excision repair (NER) of lesions from the nontranscribed regions of the genome and suggest a role for Elc1 in increasing the proficiency of repair of nontranscribed DNA, where as a component of the Rad7-Rad16-Elc1 ubiquitin ligase, it would promote the efficient turnover of the NER ensemble from the lesion site in a Rad23-19S proteasomal complex-dependent reaction. PMID- 16705156 TI - Protein phosphatase 2A stabilizes human securin, whose phosphorylated forms are degraded via the SCF ubiquitin ligase. AB - Sister chromatid segregation is triggered at the metaphase-to-anaphase transition by the activation of the protease separase. For most of the cell cycle, separase activity is kept in check by its association with the inhibitory chaperone securin. Activation of separase occurs at anaphase onset, when securin is targeted for destruction by the anaphase-promoting complex or cyclosome E3 ubiquitin protein ligase. This results in the release of the cohesins from chromosomes, which in turn allows the segregation of sister chromatids to opposite spindle poles. Here we show that human securin (hSecurin) forms a complex with enzymatically active protein phosphatase 2A (PP2A) and that it is a substrate of the phosphatase, both in vitro and in vivo. Treatment of cells with okadaic acid, a potent inhibitor of PP2A, results in various hyperphosphorylated forms of hSecurin which are extremely unstable, due to the action of the Skp1/Cul1/F-box protein complex ubiquitin ligase. We propose that PP2A regulates hSecurin levels by counteracting its phosphorylation, which promotes its degradation. Misregulation of this process may lead to the formation of tumors, in which overproduction of hSecurin is often observed. PMID- 16705155 TI - GAS41 is required for repression of the p53 tumor suppressor pathway during normal cellular proliferation. AB - GAS41 is a common subunit of the TIP60 and SRCAP complexes and is essential for cell growth and viability. Here, we report that GAS41 is required for repression of the p53 tumor suppressor pathway during normal cellular proliferation. Either GAS41 small interfering RNA-mediated knockdown of GAS41 expression or specific interruptions of the carboxy-terminal coiled-coil motif of the GAS41 protein activate the p53 tumor suppressor pathway, as evidenced by p53 up-regulation, p53 serine-15 phosphorylation, and p21 transcriptional activation. Activation of the p53 pathway does not result from changes in TIP60 complex assembly or TIP60 coactivator functions for p53, since a TIP60 complex containing a coiled-coil mutant of GAS41 retains the same composition and histone acetyltransferase activity as its wild-type counterpart and since mutant GAS41 does not compromise ectopic p53-dependent transcriptional activation in a reporter gene assay. Finally, we demonstrate that GAS41 is prebound to the promoters of two p53 tumor suppressor pathway genes (p21 and p14ARF) in normal unstressed cells but is dissociated from both promoters in response to stress signals that activate p53. Our data suggest that GAS41 plays a role in repressing the p53 tumor suppressor pathway during the normal cell cycle by a TIP60-independent mechanism. PMID- 16705157 TI - The assembly and maintenance of heterochromatin initiated by transgene repeats are independent of the RNA interference pathway in mammalian cells. AB - A role for the RNA interference (RNAi) pathway in the establishment of heterochromatin is now well accepted for various organisms. Less is known about its relevance and precise role in mammalian cells. We previously showed that tandem insertion of a 1,000-copy inducible transgene into the genome of baby hamster kidney (BHK) cells initiated the formation of an extremely condensed chromatin locus. Here, we characterized the inactive transgenic locus as heterochromatin, since it was associated with heterochromatin protein 1 (HP1), histone H3 trimethylated at lysine 9, and cytosine methylation in CpG dinucleotides. Northern blot analysis did not detect any transgene-derived small RNAs. RNAi-mediated Dicer knockdown did not disrupt the heterochromatic transgenic locus or up-regulate transgene expression. Moreover, neither Dicer knockdown nor overexpression of transgene-directed small interfering RNAs altered the bidirectional transition of the transgenic locus between the heterochromatic and euchromatic states. Interestingly, tethering of HP1 to the transgenic locus effectively induced transgene silencing and chromatin condensation in a Dicer independent manner, suggesting a role for HP1 in maintaining the heterochromatic locus. Our results suggest that the RNAi pathway is not required for the assembly and maintenance of noncentromeric heterochromatin initiated by tandem transgene repeats in mammalian cells. PMID- 16705158 TI - Insulin-like growth factor I controls a mutually exclusive association of RACK1 with protein phosphatase 2A and beta1 integrin to promote cell migration. AB - The WD repeat scaffolding protein RACK1 can mediate integration of the insulin like growth factor I receptor (IGF-IR) and integrin signaling in transformed cells. To address the mechanism of RACK1 function, we searched for regulatory proteins that associate with RACK1 in an IGF-I-dependent manner. The serine threonine phosphatase protein phosphatase 2A (PP2A) was found associated with RACK1 in serum-starved cells, and it dissociated immediately upon stimulation with IGF-I. This dissociation of PP2A from RACK1 and an IGF-I-mediated decrease in cellular PP2A activity did not occur in cells expressing either the serine 1248 or tyrosine 1250/1251 mutants of the IGF-IR that do not interact with RACK1. Recombinant RACK1 could bind to PP2A in vitro and restore phosphatase activity to PP2A from IGF-I-stimulated cells. Ligation of integrins with fibronectin or Matrigel was sufficient to facilitate IGF-I-mediated dissociation of PP2A from RACK1 and also to recruit beta1 integrin as PP2A dissociated. By using TAT-fused N-terminal and C-terminal deletion mutants of RACK1, we determined that both PP2A and beta1 integrin interact in the C terminus of RACK1 within WD repeats 4 to 7. This suggests that integrin ligation displaces PP2A from RACK1. MCF-7 cells overexpressing RACK1 exhibited enhanced motility, which could be reversed by the PP2A inhibitor okadaic acid. Small interfering RNA-mediated suppression of RACK1 also decreased the migratory capacity of DU145 cells. Taken together, our findings indicate that RACK1 enhances IGF-I-mediated cell migration through its ability to exclusively associate with either beta1 integrin or PP2A in a complex at the IGF-IR. PMID- 16705160 TI - Phosphorylation of Jak2 on Ser(523) inhibits Jak2-dependent leptin receptor signaling. AB - The leptin receptor, LRb, and other cytokine receptors are devoid of intrinsic enzymatic activity and rely upon the activity of constitutively associated Jak family tyrosine kinases to mediate intracellular signaling. In order to clarify mechanisms by which Jak2, the cognate LRb-associated Jak kinase, is regulated and mediates downstream signaling, we employed tandem mass spectroscopic analysis to identify phosphorylation sites on Jak2. We identified Ser523 as the first described site of Jak2 serine phosphorylation and demonstrated that this site is phosphorylated on Jak2 from intact cells and mouse spleen. Ser523 was highly phosphorylated in HEK293 cells independently of LRb-Jak2 activation, suggesting a potential role for the phosphorylation of Ser523 in the regulation of LRb by other pathways. Indeed, mutation of Ser523 sensitized and prolonged signaling by Jak2 following activation by the intracellular domain of LRb. The effect of Ser523 on Jak2 function was independent of Tyr570-mediated inhibition. Thus, the phosphorylation of Jak2 on Ser523 inhibits Jak2 activity and represents a novel mechanism for the regulation of Jak2-dependent cytokine signaling. PMID- 16705159 TI - Phosphorylation of JAK2 at serine 523: a negative regulator of JAK2 that is stimulated by growth hormone and epidermal growth factor. AB - The tyrosine kinase JAK2 is a key signaling protein for at least 20 receptors in the cytokine/hematopoietin receptor superfamily and is a component of signaling for multiple receptor tyrosine kinases and several G-protein-coupled receptors. In this study, phosphopeptide affinity enrichment and mass spectrometry identified serine 523 (Ser523) in JAK2 as a site of phosphorylation. A phosphoserine 523 antibody revealed that Ser523 is rapidly but transiently phosphorylated in response to growth hormone (GH). MEK1 inhibitor UO126 suppresses GH-dependent phosphorylation of Ser523, suggesting that extracellular signal-regulated kinases (ERKs) 1 and/or 2 or another kinase downstream of MEK1 phosphorylate Ser523 in response to GH. Other ERK activators, phorbol 12 myristate 13-acetate and epidermal growth factor, also stimulate phosphorylation of Ser523. When Ser523 in JAK2 was mutated, JAK2 kinase activity as well as GH dependent tyrosyl phosphorylation of JAK2 and Stat5 was enhanced, suggesting that phosphorylation of Ser523 inhibits JAK2 kinase activity. We hypothesize that phosphorylation of Ser523 in JAK2 by ERKs 1 and/or 2 or other as-yet-unidentified kinases acts in a negative feedback manner to dampen activation of JAK2 in response to GH and provides a mechanism by which prior exposure to environmental factors that regulate Ser523 phosphorylation might modulate the cell's response to GH. PMID- 16705161 TI - Genesis of and trafficking to the Maurer's clefts of Plasmodium falciparum infected erythrocytes. AB - Malaria parasites export proteins beyond their own plasma membrane to locations in the red blood cells in which they reside. Maurer's clefts are parasite-derived structures within the host cell cytoplasm that are thought to function as a sorting compartment between the parasite and the erythrocyte membrane. However, the genesis of this compartment and the signals directing proteins to the Maurer's clefts are not known. We have generated Plasmodium falciparum-infected erythrocytes expressing green fluorescent protein (GFP) chimeras of a Maurer's cleft resident protein, the membrane-associated histidine-rich protein 1 (MAHRP1). Chimeras of full-length MAHRP1 or fragments containing part of the N terminal domain and the transmembrane domain are successfully delivered to Maurer's clefts. Other fragments remain trapped within the parasite. Fluorescence photobleaching and time-lapse imaging techniques indicate that MAHRP1-GFP is initially trafficked to isolated subdomains in the parasitophorous vacuole membrane that appear to represent nascent Maurer's clefts. The data suggest that the Maurer's clefts bud from the parasitophorous vacuole membrane and diffuse within the erythrocyte cytoplasm before taking up residence at the cell periphery. PMID- 16705162 TI - Sgs1 regulates gene conversion tract lengths and crossovers independently of its helicase activity. AB - RecQ helicases maintain genome stability and suppress tumors in higher eukaryotes through roles in replication and DNA repair. The yeast RecQ homolog Sgs1 interacts with Top3 topoisomerase and Rmi1. In vitro, Sgs1 binds to and branch migrates Holliday junctions (HJs) and the human RecQ homolog BLM, with Top3alpha, resolves synthetic double HJs in a noncrossover sense. Sgs1 suppresses crossovers during the homologous recombination (HR) repair of DNA double-strand breaks (DSBs). Crossovers are associated with long gene conversion tracts, suggesting a model in which Sgs1 helicase catalyzes reverse branch migration and convergence of double HJs for noncrossover resolution by Top3. Consistent with this model, we show that allelic crossovers and gene conversion tract lengths are increased in sgs1Delta. However, crossover and tract length suppression was independent of Sgs1 helicase activity, which argues against helicase-dependent HJ convergence. HJs may converge passively by a "random walk," and Sgs1 may play a structural role in stimulating Top3-dependent resolution. In addition to the new helicase independent functions for Sgs1 in crossover and tract length control, we define three new helicase-dependent functions, including the suppression of chromosome loss, chromosome missegregation, and synthetic lethality in srs2Delta. We propose that Sgs1 has helicase-dependent functions in replication and helicase independent functions in DSB repair by HR. PMID- 16705163 TI - SWI/SNF binding to the HO promoter requires histone acetylation and stimulates TATA-binding protein recruitment. AB - We use chromatin immunoprecipitation assays to show that the Gcn5 histone acetyltransferase in SAGA is required for SWI/SNF association with the HO promoter and that binding of SWI/SNF and SAGA are interdependent. Previous results showed that SWI/SNF binding to HO was Gcn5 independent, but that work used a strain with a mutation in the Ash1 daughter-specific repressor of HO expression. Here, we show that Ash1 functions as a repressor that inhibits SWI/SNF binding and that Gcn5 is required to overcome Ash1 repression in mother cells to allow HO transcription. Thus, Gcn5 facilitates SWI/SNF binding by antagonizing Ash1. Similarly, a mutation in SIN3, like an ash1 mutation, allows both HO expression and SWI/SNF binding in the absence of Gcn5. Although Ash1 has recently been identified in a Sin3-Rpd3 complex, our genetic analysis shows that Ash1 and Sin3 have distinct functions in regulating HO. Analysis of mutant strains shows that SWI/SNF binding and HO expression are correlated and regulated by histone acetylation. The defect in HO expression caused by a mutant SWI/SNF with a Swi2(E834K) substitution can be partially suppressed by ash1 or spt3 mutation or by a gain-of-function V71E substitution in the TATA-binding protein (TBP). Spt3 inhibits TBP binding at HO, and genetic analysis suggests that Spt3 and TBP(V71E) act in the same pathway, distinct from that of Ash1. We have detected SWI/SNF binding at the HO TATA region, and our results suggest that SWI/SNF, either directly or indirectly, facilitates TBP binding at HO. PMID- 16705164 TI - Two-step regulation of Ad4BP/SF-1 gene transcription during fetal adrenal development: initiation by a Hox-Pbx1-Prep1 complex and maintenance via autoregulation by Ad4BP/SF-1. AB - The orphan nuclear receptor Ad4BP/SF-1 (adrenal 4 binding protein/steroidogenic factor 1) is essential for the proper development and function of reproductive and steroidogenic tissues. Although the expression of Ad4BP/SF-1 is specific for those tissues, the mechanisms underlying this tissue-specific expression remain unknown. In this study, we used transgenic mouse assays to examine the regulation of the tissue-specific expression of Ad4BP/SF-1. An investigation of the entire Ad4BP/SF-1 gene locus revealed a fetal adrenal enhancer (FAdE) in intron 4 containing highly conserved binding sites for Pbx-Prep, Pbx-Hox, and Ad4BP/SF-1. Transgenic assays revealed that the Ad4 sites, together with Ad4BP/SF-1, develop an autoregulatory loop and thereby maintain transcription, while the Pbx/Prep and Pbx/Hox sites initiate transcription prior to the establishment of the autoregulatory loop. Indeed, a limited number of Hox family members were found to be expressed in the adrenal primordia. Whether a true fetal-type adrenal cortex is present in mice remained controversial, and this argument was complicated by the postnatal development of the so-called X zone. Using transgenic mice with lacZ driven by the FAdE, we clearly identified a fetal adrenal cortex in mice, and the X zone is the fetal adrenal cells accumulated at the juxtamedullary region after birth. PMID- 16705166 TI - MAL and ternary complex factor use different mechanisms to contact a common surface on the serum response factor DNA-binding domain. AB - The transcription factor serum response factor (SRF) interacts with its cofactor, MAL/MKL1, a member of the myocardin-related transcription factor (MRTF) family, through its DNA-binding domain. We define a seven-residue sequence within the conserved MAL B1 region essential and sufficient for complex formation. The neighboring Q-box sequence facilitates this interaction. The B1 and Q-box regions also have antagonistic effects on MAL nuclear import, but the residues involved are largely distinct. Both MAL and the ternary complex factor (TCF) family of SRF cofactors interact with a hydrophobic groove and pocket on the SRF DNA-binding domain. Unlike the TCFs, however, interaction of MAL with SRF is impaired by SRF alphaI-helix mutations that reduce DNA bending in the SRF-DNA complex. A clustered SRF alphaI-helix mutation strongly impairs MAL-SRF complex formation but does not affect DNA distortion in the MAL-SRF complex. MAL-SRF complex formation is facilitated by DNA binding. DNase I footprinting indicates that in the SRF-MAL complex MAL directly contacts DNA. These contacts, which flank the DNA sequences protected from DNase I by SRF, are required for effective MAL-SRF complex formation in gel mobility shift assays. We propose a model of MAL-SRF complex formation in which MAL interacts with SRF by the addition of a beta strand to the SRF DNA-binding domain beta-sheet region, while SRF-induced DNA bending facilitates MAL-DNA contact. PMID- 16705165 TI - Role of Doa1 in the Saccharomyces cerevisiae DNA damage response. AB - The cellular response to DNA damage requires not only direct repair of the damage but also changes in the DNA replication machinery, chromatin, and transcription that facilitate survival. Here, we describe Saccharomyces cerevisiae Doa1, which helps to control the damage response by channeling ubiquitin from the proteosomal degradation pathway into pathways that mediate altered DNA replication and chromatin modification. DOA1 interacts with genes involved in PCNA ubiquitination, including RAD6, RAD18, RAD5, UBC13, and MMS2, as well as genes involved in histone H2B ubiquitination or deubiquitination, including RAD6, BRE1, LGE1, CDC73, UBP8, UBP10, and HTB2. In the absence of DOA1, damage-induced ubiquitination of PCNA does not occur. In addition, the level of ubiquitinated H2B is decreased under normal conditions and completely absent in the presence of DNA damage. In the case of PCNA, the defect associated with the doa1Delta mutant is alleviated by overexpression of ubiquitin, but in the case of H2B, it is not. The data suggest that Doa1 is the major source of ubiquitin for the DNA damage response and that Doa1 also plays an additional essential and more specific role in the monoubiquitination of histone H2B. PMID- 16705167 TI - T-cell protein tyrosine phosphatase (Tcptp) is a negative regulator of colony stimulating factor 1 signaling and macrophage differentiation. AB - Mice null for the T-cell protein tyrosine phosphatase (Tcptp-/-) die shortly after birth due to complications arising from the development of a systemic inflammatory disease. It was originally reported that Tcptp-/- mice have increased numbers of macrophages in the spleen; however, the mechanism underlying the aberrant growth and differentiation of macrophages in Tcptp-/- mice is not known. We have identified Tcptp as an important regulator of colony-stimulating factor 1 (CSF-1) signaling and mononuclear phagocyte development. The number of CSF-1-dependent CFU is increased in Tcptp-/- bone marrow. Tcptp-/- mice also have increased numbers of granulocyte-macrophage precursors (GMP), and these Tcptp-/- GMP yield more macrophage colonies in response to CSF-1 relative to wild-type cells. Furthermore, we have identified the CSF-1 receptor (CSF-1R) as a physiological target of Tcptp through substrate-trapping experiments and its hyperphosphorylation in Tcptp-/- macrophages. Tcptp-/- macrophages also have increased tyrosine phosphorylation and recruitment of a Grb2/Gab2/Shp2 complex to the CSF-1R and enhanced activation of Erk after CSF-1 stimulation, which are important molecular events in CSF-1-induced differentiation. These data implicate Tcptp as a critical regulator of CSF-1 signaling and mononuclear phagocyte development in hematopoiesis. PMID- 16705168 TI - Prohibitin facilitates cellular senescence by recruiting specific corepressors to inhibit E2F target genes. AB - Prohibitin is a growth regulatory gene that has pleiotropic functions in the nucleus, mitochondria, and cytoplasmic compartments. Earlier studies had proposed a role for prohibitin in modulating cellular senescence, but the underlying mechanisms remain unknown. Here we show that senescence induced by DNA-damaging agents causes the localization of prohibitin to specific heterochromatic foci. Prohibitin could bind to heterochromatin protein 1 (HP1) family proteins and colocalized with HP1gamma in senescence-associated heterochromatic foci. Further, HP1gamma could synergize with prohibitin to repress E2F1-mediated transcriptional activity. The depletion of prohibitin by small interfering RNA or antisense techniques led to a reduction in the senescent phenotype, correlating with a reduced expression of senescence-associated beta-galactosidase and fewer numbers of senescence-associated heterochromatic foci. Chromatin immunoprecipitation assays showed that prohibitin is needed for the recruitment of HP1gamma to E2F1 regulated proliferative promoters, leading to their repression. The ablation of prohibitin prevented the recruitment of HPIgamma, but not Suv39H, to the promoters upon senescence. Prohibitin-mediated recruitment of HP1gamma occurred in only senescent cells, not in quiescent cells; thus, there is a dichotomy in the recruitment of different corepressors by prohibitin, depending on the type of growth arrest. These studies show that prohibitin plays a vital role in inducing cellular senescence. PMID- 16705169 TI - Cathepsin L stabilizes the histone modification landscape on the Y chromosome and pericentromeric heterochromatin. AB - Posttranslational histone modifications and histone variants form a unique epigenetic landscape on mammalian chromosomes where the principal epigenetic heterochromatin markers, trimethylated histone H3(K9) and the histone H2A.Z, are inversely localized in relation to each other. Trimethylated H3(K9) marks pericentromeric constitutive heterochromatin and the male Y chromosome, while H2A.Z is dramatically reduced at these chromosomal locations. Inactivation of a lysosomal and nuclear protease, cathepsin L, causes a global redistribution of epigenetic markers. In cathepsin L knockout cells, the levels of trimethylated H3(K9) decrease dramatically, concomitant with its relocation away from heterochromatin, and H2A.Z becomes enriched at pericentromeric heterochromatin and the Y chromosome. This change is also associated with global relocation of heterochromatin protein HP1 and histone H3 methyltransferase Suv39h1 away from constitutive heterochromatin; however, it does not affect DNA methylation or chromosome segregation, phenotypes commonly associated with impaired histone H3(K9) methylation. Therefore, the key constitutive heterochromatin determinants can dynamically redistribute depending on physiological context but still maintain the essential function(s) of chromosomes. Thus, our data show that cathepsin L stabilizes epigenetic heterochromatin markers on pericentromeric heterochromatin and the Y chromosome through a novel mechanism that does not involve DNA methylation or affect heterochromatin structure and operates on both somatic and sex chromosomes. PMID- 16705170 TI - N-cadherin and keratinocyte growth factor receptor mediate the functional interplay between Ki-RASG12V and p53V143A in promoting pancreatic cell migration, invasion, and tissue architecture disruption. AB - The genetic basis of pancreatic ductal adenocarcinoma, which constitutes the most common type of pancreatic malignancy, involves the sequential activation of oncogenes and inactivation of tumor suppressor genes. Among the pivotal genetic alterations are Ki-RAS oncogene activation and p53 tumor suppressor gene inactivation. We explain that the combination of these genetic events facilitates pancreatic carcinogenesis as revealed in novel three-dimensional cell (spheroid cyst) culture and in vivo subcutaneous and orthotopic xenotransplantation models. N-cadherin, a member of the classic cadherins important in the regulation of cell cell adhesion, is induced in the presence of Ki-RAS mutation but subsequently downregulated with the acquisition of p53 mutation as revealed by gene microarrays and corroborated by reverse transcription-PCR and Western blotting. N cadherin modulates the capacity of pancreatic ductal cells to migrate and invade, in part via complex formation with keratinocyte growth factor receptor and neural cell adhesion molecule and in part via interaction with p120-catenin. However, modulation of these complexes by Ki-RAS and p53 leads to enhanced cell migration and invasion. This preferentially induces the downstream effector AKT over mitogen-activated protein kinase to execute changes in cellular behavior. Thus, we are able to define molecules that in part are directly affected by Ki-RAS and p53 during pancreatic ductal carcinogenesis, and this provides a platform for potential new molecularly based therapeutic interventions. PMID- 16705171 TI - Focal adhesion kinase controls cellular levels of p27/Kip1 and p21/Cip1 through Skp2-dependent and -independent mechanisms. AB - Endothelial cell proliferation is a critical step in angiogenesis and requires a coordinated response to soluble growth factors and the extracellular matrix. As focal adhesion kinase (FAK) integrates signals from both adhesion events and growth factor stimulation, we investigated its role in endothelial cell proliferation. Expression of a dominant-negative FAK protein, FAK-related nonkinase (FRNK), impaired phosphorylation of FAK and blocked DNA synthesis in response to multiple angiogenic stimuli. These results coincided with elevated cyclin-dependent kinase inhibitors (CDKIs) p21/Cip and p27/Kip, as a consequence of impaired degradation. FRNK inhibited the expression of Skp2, an F-box protein that targets CDKIs, by inhibiting mitogen-induced mRNA. The FAK-regulated degradation of p27/Kip was Skp2 dependent, while levels of p21/Cip were regulated independent of Skp2. Skp2 is required for endothelial cell proliferation as a consequence of degrading p27. Finally, knockdown of both p21 and p27 in FRNK expressing cells completely restored mitogen-induced endothelial cell proliferation. These data demonstrate a critical role for FAK in the regulation of CDKIs through two independent mechanisms: Skp2 dependent and Skp2 independent. They also provide important insights into the requirement of focal adhesion kinase for normal vascular development and reveal novel regulatory control points for angiogenesis. PMID- 16705172 TI - Novel lymphotoxin alpha (LTalpha) knockout mice with unperturbed tumor necrosis factor expression: reassessing LTalpha biological functions. AB - Lymphotoxin alpha (LTalpha) can exist in soluble form and exert tumor necrosis factor (TNF)-like activity through TNF receptors. Based on the phenotypes of knockout (KO) mice, the physiological functions of LTalpha and TNF are considered partly redundant, in particular, in supporting the microarchitecture of the spleen and in host defense. We exploited Cre-LoxP technology to generate a novel neomycin resistance gene (neo) cassette-free LTalpha-deficient mouse strain (neo free LTalpha KO [LTalphaDelta/Delta]). Unlike the "conventional" LTalpha-/- mice, new LTalphaDelta/Delta animals were capable of producing normal levels of systemic TNF upon lipopolysaccharide (LPS) challenge and were susceptible to LPS/D-galactosamine (D-GalN) toxicity. Activated neutrophils, monocytes, and macrophages from LTalphaDelta/Delta mice expressed TNF normally at both the mRNA and protein levels as opposed to conventional LTalpha KO mice, which showed substantial decreases in TNF. Additionally, the spleens of the neo-free LTalpha KO mice displayed several features resembling those of LTbeta KO mice rather than conventional LTalpha KO animals. The phenotype of the new LTalphaDelta/Delta mice indicates that LTalpha plays a smaller role in lymphoid organ maintenance than previously thought and has no direct role in the regulation of TNF expression. PMID- 16705173 TI - A conserved Myc protein domain, MBIV, regulates DNA binding, apoptosis, transformation, and G2 arrest. AB - The myc family of oncogenes is well conserved throughout evolution. Here we present the characterization of a domain conserved in c-, N-, and L-Myc from fish to humans, N-Myc317-337, designated Myc box IV (MBIV). A deletion of this domain leads to a defect in Myc-induced apoptosis and in some transformation assays but not in cell proliferation. Unlike other Myc mutants, MycDeltaMBIV is not a simple loss-of-function mutant because it is hyperactive for G2 arrest in primary cells. Microarray analysis of genes regulated by N-MycDeltaMBIV reveals that it is weakened for transactivation and repression but not nearly as defective as N MycDeltaMBII. Although the mutated region is not part of the previously defined DNA binding domain, we find that N-MycDeltaMBIV has a significantly lower affinity for DNA than the wild-type protein in vitro. Furthermore, chromatin immunoprecipitation shows reduced binding of N-MycDeltaMBIV to some target genes in vivo, which correlates with the defect in transactivation. Thus, this conserved domain has an unexpected role in Myc DNA binding activity. These data also provide a novel separation of Myc functions linked to the modulation of DNA binding activity. PMID- 16705175 TI - Deletion of SERP1/RAMP4, a component of the endoplasmic reticulum (ER) translocation sites, leads to ER stress. AB - Stress-associated endoplasmic reticulum (ER) protein 1 (SERP1), also known as ribosome-associated membrane protein 4 (RAMP4), is a Sec61-associated polypeptide that is induced by ER stress. SERP1-/- mice, made by targeted gene disruption, demonstrated growth retardation, increased mortality, and impaired glucose tolerance. Consistent with high levels of SERP1 expression in pancreas, pancreatic islets from SERP1-/- mice failed to rapidly synthesize proinsulin in response to a glucose load. In addition, reduced size and enhanced ER stress were observed in the anterior pituitary of SERP1-/- mice, and growth hormone production was slowed in SERP1-/- pituitary after insulin stimulation. Experiments using pancreatic microsomes revealed aberrant association of ribosomes and the Sec61 complex and enhanced ER stress in SERP1-/- pancreas. In basal conditions, the Sec61 complex in SERP1-/- microsomes was more cofractionated with ribosomes, compared with SERP1+/+ counterparts, in high-salt conditions. In contrast, after glucose stimulation, the complex showed less cofractionation at an early phase (45 min) but more at a later phase (120 min). Although intracellular insulin/proinsulin levels were not significantly changed in both genotypes, these results suggest that subtle changes in translocation efficiency play an important role in the regulation of ER stress and rapid polypeptide synthesis. PMID- 16705176 TI - GLUT8 is dispensable for embryonic development but influences hippocampal neurogenesis and heart function. AB - GLUT8 is a glucose transporter isoform expressed at high levels in testis; at intermediate levels in the brain, including the hippocampus; and at lower levels in the heart and several other tissues. GLUT8 is located in an intracellular compartment and does not appear to translocate to the cell surface, except in blastocysts, where insulin has been reported to induce its surface expression. Here, we generated mice with inactivation of the glut8 gene. We showed that expression of GLUT8 was not required for normal embryonic development and that glut8-/- mice had normal postnatal development, glucose homeostasis, and response to mild stress. Adult glut8-/- mice showed increased proliferation of hippocampal cells but no defect in memory acquisition and retention. Absence of GLUT8 from the heart did not alter heart size and morphology but led to an increase in P wave duration, which was not associated with abnormal Nav1.5 Na+ channel or connexin expression. Thus, absence of GLUT8 expression in the mouse caused complex but mild physiological alterations. PMID- 16705174 TI - Cyclin D1 regulates cellular migration through the inhibition of thrombospondin 1 and ROCK signaling. AB - Cyclin D1 is overexpressed in human tumors, correlating with cellular metastasis, and is induced by activating Rho GTPases. Herein, cyclin D1-deficient mouse embryo fibroblasts (MEFs) exhibited increased adhesion and decreased motility compared with wild-type MEFs. Retroviral transduction of cyclin D1 reversed these phenotypes. Mutational analysis of cyclin D1 demonstrated that its effects on cellular adhesion and migration were independent of the pRb and p160 coactivator binding domains. Genomewide expression arrays identified a subset of genes regulated by cyclin D1, including Rho-activated kinase II (ROCKII) and thrombospondin 1 (TSP-1). cyclin D1(-/-) cells showed increased Rho GTP and ROCKII activity and signaling, with increased phosphorylation of LIM kinase, cofilin (Ser3), and myosin light chain 2 (Thr18/Ser19). Cyclin D1 repressed ROCKII and TSP-1 expression, and the migratory defect of cyclin D1(-/-) cells was reversed by ROCK inhibition or TSP-1 immunoneutralizing antibodies. cyclin E knockin to the cyclin D1(-/-) MEFs rescued the DNA synthesis defect of cyclin D1( /-) MEFs but did not rescue either the migration defect or the abundance of ROCKII. Cyclin D1 promotes cellular motility through inhibiting ROCK signaling and repressing the metastasis suppressor TSP-1. PMID- 16705177 TI - Translational control by neuroguidin, a eukaryotic initiation factor 4E and CPEB binding protein. AB - CPEB-mediated translation is important in early development and neuronal synaptic plasticity. Here, we describe a new eukaryotic initiation factor 4E (eIF4E) binding protein, Neuroguidin (Ngd), and its interaction with CPEB. In the mammalian nervous system, Ngd is detected as puncta in axons and dendrites and in growth cones and filopodia. Ngd contains three motifs that resemble those present in eIF4G, 4EBP, Cup, and Maskin, all of which are eIF4E binding proteins. Ngd binds eIF4E directly, and all three motifs must be deleted to abrogate the interaction between these two proteins. In injected Xenopus oocytes, Ngd binds CPEB and, most importantly, represses translation in a cytoplasmic polyadenylation element (CPE)-dependent manner. In Xenopus embryos, Ngd is found in both neural tube and neural crest cells. The injection of morpholino containing antisense oligonucleotides directed against ngd mRNA disrupts neural tube closure and neural crest migration; however, the wild-type phenotype is restored by the injection of a rescuing ngd mRNA. These data suggest that Ngd guides neural development by regulating the translation of CPE-containing mRNAs. PMID- 16705179 TI - Embryonic fibroblasts from mice lacking Tgif were defective in cell cycling. AB - Holoprosencephaly (HPE) is the most common structural anomaly of the human brain, resulting from incomplete cleavage of the developing forebrain during embryogenesis. Haploinsufficient mutations in the TG-interacting factor (TGIF) gene were previously identified in a subset of HPE families and sporadic patients, and this gene is located within a region of chromosome 18 that is associated with nonrandom chromosomal aberrations in HPE patients. TGIF is a three-amino-acid loop extension (TALE) homeodomain-containing transcription factor that functions both as a corepressor of the transforming growth factor beta (TGF-beta) pathway and as a competitor of the retinoic acid pathway. Here we describe mice deficient in Tgif that exhibited laterality defects and growth retardation and developed kinked tails. Cellular analysis of mutant mouse embryonic fibroblasts (MEFs) demonstrated for the first time that Tgif regulates proliferation and progression through the G1 cell cycle phase. Additionally, wild type human TGIF was able to rescue this proliferative defect in MEFs. In contrast, a subset of human Tgif mutations detected in HPE patients was unable to rescue the proliferative defect. However, an absence of Tgif did not alter the normal inhibition of proliferation caused by treatment with TGF-beta or retinoic acid. Developmental control of proliferation by Tgif may play a role in the pathogenesis of HPE. PMID- 16705178 TI - A nuclear export signal and phosphorylation regulate Dok1 subcellular localization and functions. AB - Dok1 is believed to be a mainly cytoplasmic adaptor protein which down-regulates mitogen-activated protein kinase activation, inhibits cell proliferation and transformation, and promotes cell spreading and cell migration. Here we show that Dok1 shuttles between the nucleus and cytoplasm. Treatment of cells with leptomycin B (LMB), a specific inhibitor of the nuclear export signal (NES) dependent receptor CRM1, causes nuclear accumulation of Dok1. We have identified a functional NES (348LLKAKLTDPKED359) that plays a major role in the cytoplasmic localization of Dok1. Src-induced tyrosine phosphorylation prevented the LMB mediated nuclear accumulation of Dok1. Dok1 cytoplasmic localization is also dependent on IKKbeta. Serum starvation or maintaining cells in suspension favor Dok1 nuclear localization, while serum stimulation, exposure to growth factor, or cell adhesion to a substrate induce cytoplasmic localization. Functionally, nuclear NES-mutant Dok1 had impaired ability to inhibit cell proliferation and to promote cell spreading and cell motility. Taken together, our results provide the first evidence that Dok1 transits through the nucleus and is actively exported into the cytoplasm by the CRM1 nuclear export system. Nuclear export modulated by external stimuli and phosphorylation may be a mechanism by which Dok1 is maintained in the cytoplasm and membrane, thus regulating its signaling functions. PMID- 16705180 TI - Congenital hypothyroidism (cretinism) in neuroD2-deficient mice. AB - Mice lacking neuroD2, a basic helix-loop-helix transcription factor involved in brain development, show growth retardation and other abnormalities consistent with hypothalamic-pituitary-thyroid (HPT) axis dysfunction. neuroD2 is expressed in the paraventricular hypothalamic nuclei, the anterior lobe of pituitary, and the thyroid gland. In neuroD2-deficient mice, thyrotropin-releasing hormone, thyroid-stimulating hormone, and thyroid hormone are decreased in these three regions, respectively. neuroD2-null mice typically die 2 to 3 weeks after birth, but those treated with replacement doses of thyroxine survived more than 8 weeks. These data indicate that neuroD2 is expressed throughout the HPT axis and that all levels of the axis are functionally affected by its absence in mice. PMID- 16705181 TI - Multisite protein kinase A and glycogen synthase kinase 3beta phosphorylation leads to Gli3 ubiquitination by SCFbetaTrCP. AB - Gli3 is a zinc finger transcription factor proteolytically processed into a truncated repressor lacking C-terminal activation domains. Gli3 processing is stimulated by protein kinase A (PKA) and inhibited by Hedgehog signaling, a major signaling pathway in vertebrate development and disease. We show here that multisite glycogen synthase kinase 3beta (GSK3beta) phosphorylation and ubiquitination by SCFbetaTrCP are required for Gli3 processing. We identified multiple betaTrCP-binding sites related to the DSGX2-4S motif in Gli3, which are intertwined with PKA and GSK3beta sites, and SCFbetaTrCP target lysines that are essential for processing. Our results support a simple model whereby PKA triggers a cascade of Gli3 phosphorylation by GSK3beta and CK1 that leads to direct betaTrCP binding and ubiquitination by SCFbetaTrCP. Binding of betaTrCP to Gli3 N and C-terminal domains lacking DSGX2-4S-related motifs was also observed, which could reflect indirect interaction via other components of Hedgehog signaling, such as the tumor suppressor Sufu. Gli3 therefore joins a small set of transcription factors whose processing is regulated by the ubiquitin-proteasome pathway. Our study sheds light on the role of PKA phosphorylation in Gli3 processing and will help to analyze how dose-dependent tuning of Gli3 processing is achieved by Hedgehog signaling. PMID- 16705182 TI - Phosphorylation of amyloid precursor protein (APP) at Thr668 regulates the nuclear translocation of the APP intracellular domain and induces neurodegeneration. AB - Amyloid precursor protein (APP) has eight potential phosphorylation sites in its cytoplasmic domain. Recently, it has demonstrated that the constitutive phosphorylation of APP at T668 (APP695 isoform numbering) was observed specifically in the brain. Neuron-specific phosphorylation of APP at T668 is thought to be important for neuronal functions of APP, although its exact physiological significance remains to be clarified. In this study, we show that the phosphorylation of the APP intracellular domain (AICD) at T668 is essential for its binding to Fe65 and its nuclear translocation and affects the resultant neurotoxicity, possibly mediated through the induction of glycogen synthase kinase 3beta and tau phosphorylation by enhancing the formation of a ternary complex with Fe65 and CP2 transcription factor. Taken together, these results suggest that the phosphorylation of AICD at T668 contributes to the neuronal degeneration in Alzheimer's disease (AD) by regulating its translocation into the nucleus and then affects neurodegeneration; therefore, the specific inhibitor of T668 phosphorylation might be the target of AD therapy. PMID- 16705183 TI - p14ARF activates a Tip60-dependent and p53-independent ATM/ATR/CHK pathway in response to genotoxic stress. AB - p14ARF is a tumor suppressor that controls a well-described p53/Mdm2-dependent checkpoint in response to oncogenic signals. Here, new insights into the tumor suppressive function of p14ARF are provided. We previously showed that p14ARF can induce a p53-independent G2 cell cycle arrest. In this study, we demonstrate that the activation of ATM/ATR/CHK signaling pathways contributes to this G2 checkpoint and highlight the interrelated roles of p14ARF and the Tip60 protein in the initiation of this DNA damage-signaling cascade. We show that Tip60 is a new direct p14ARF binding partner and that its expression is upregulated and required for ATM/CHK2 activation in response to p14ARF. Strikingly, both p14ARF and Tip60 products accumulate following a cell treatment with alkylating agents and are absolutely required for ATM/CHK2 activation in this setting. Moreover, and consistent with p14ARF being a determinant of CHK2 phosphorylation in lung carcinogenesis, a strong correlation between p14ARF and phospho-CHK2 (Thr68) protein expression is observed in human lung tumors (P < 0.00006). Overall, these data point to a novel regulatory pathway that mediates the p53-independent negative-cell-growth control of p14ARF. Inactivation of this pathway is likely to contribute to lung carcinogenesis. PMID- 16705184 TI - E Proteins and Id2 converge on p57Kip2 to regulate cell cycle in neural cells. AB - A precise balance between proliferation and differentiation must be maintained during neural development to obtain the correct proportion of differentiated cell types in the adult nervous system. The basic helix-loop-helix (bHLH) transcription factors known as E proteins and their natural inhibitors, the Id proteins, control the timing of differentiation and terminal exit from the cell cycle. Here we show that progression into S phase of human neuroblastoma cells is prevented by E proteins and promoted by Id2. Cyclin-dependent kinase inhibitors (CKI) have been identified as key effectors of cell cycle arrest in differentiating cells. However, p57Kip2 is the only CKI that is absolutely required for normal development. Through the use of global gene expression analysis in neuroblastoma cells engineered to acutely express the E protein E47 and Id2, we find that p57Kip2 is a target of E47. Consistent with the role of Id proteins, Id2 prevents activation of p57Kip2 expression, and the retinoblastoma tumor suppressor protein, a known Id2 inhibitor, counters this activity. The strong E47-mediated inhibition of entry into S phase is entirely reversed in cells in which expression of p57Kip2 is silenced by RNA interference. During brain development, expression of p57Kip2 is opposite that of Id2. Our findings identify p57Kip2 as a functionally relevant target recruited by bHLH transcription factors to induce cell cycle arrest in developing neuroblasts and suggest that deregulated expression of Id proteins may be an epigenetic mechanism to silence expression of this CKI in neural tumors. PMID- 16705185 TI - Thoc1/Hpr1/p84 is essential for early embryonic development in the mouse. AB - The yeast TREX complex physically couples elongating RNA polymerase II with RNA processing and nuclear RNA export factors to facilitate regulated gene expression. Hpr1p is an essential component of TREX, and loss of Hpr1p compromises transcriptional elongation, RNA export, and genome stability. Despite these defects, HPR1 is not essential for viability in yeast. A functional orthologue of Hpr1p has been identified in metazoan species and is variously known as Thoc1, Hpr1, or p84. However, the physiological functions of this protein have not been determined. Here, we describe the generation and phenotypic characterization of mice containing a null allele of the Thoc1 gene. Heterozygous null Thoc1 mice are born at the expected Mendelian frequency with no phenotype distinguishable from the wild type. In contrast, homozygous null mice are not recovered, indicating that Thoc1 is required for embryonic development. Embryonic development is arrested around the time of implantation, as blastocysts exhibit hatching and blastocyst outgrowth defects upon in vitro culture. Cells of the inner cell mass are particularly dependent on Thoc1, as these cells rapidly lose viability coincident with Thoc1 protein loss. While Hpr1p is not essential for the viability of unicellular yeasts, the orthologous Thoc1 protein is required for viability of the early mouse embryo. PMID- 16705187 TI - The transcription elongation complex directs activation-induced cytidine deaminase-mediated DNA deamination. AB - Activation-induced cytidine deaminase (AID) is a single-stranded DNA deaminase required for somatic hypermutation of immunoglobulin (Ig) genes, a key process in the development of adaptive immunity. Transcription provides a single-stranded DNA substrate for AID, both in vivo and in vitro. We present here an assay which can faithfully replicate all of the molecular features of the initiation of hypermutation of Ig genes in vivo. In this assay, which detects AID-mediated deamination in the context of transcription by Escherichia coli RNA polymerase, deamination targets either strand and declines in efficiency as the distance from the promoter increases. We show that AID binds DNA exposed by the transcribing polymerase, implicating the polymerase itself as the vehicle which distributes AID on DNA as it moves away from the promoter. PMID- 16705188 TI - Biomarker discovery by imaging mass spectrometry: transthyretin is a biomarker for gentamicin-induced nephrotoxicity in rat. AB - Adverse drug effects are often associated with pathological changes in tissue. An accurate depiction of the undesired affected area, possibly supported by mechanistic data, is important to classify the effects with regard to relevance for human patients. MALDI imaging MS represents a new analytical tool to directly provide the spatial distribution and the relative abundance of proteins in tissue. Here we evaluate this technique to investigate potential toxicity biomarkers in kidneys of rats that were administered gentamicin, a well known nephrotoxicant. Differential analysis of the mass spectrum profiles revealed a spectral feature at 12,959 Da that strongly correlates with histopathology alterations of the kidney. We unambiguously identified this spectral feature as transthyretin (Ser(28)-Gln(146)) using an innovative combination of tissue microextraction and fractionation by reverse-phase liquid chromatography followed by a top-down tandem mass spectrometric approach. Our findings clearly demonstrate the emerging role of imaging MS in the discovery of toxicity biomarkers and in obtaining mechanistic insights concerning toxicity mechanisms. PMID- 16705186 TI - Alterations in expression and chromatin configuration of the alpha hemoglobin stabilizing protein gene in erythroid Kruppel-like factor-deficient mice. AB - Erythroid Kruppel-like factor (EKLF) is an erythroid zinc finger protein identified by its interaction with a CACCC sequence in the beta-globin promoter, where it establishes local chromatin structure permitting beta-globin gene transcription. We sought to identify other EKLF target genes and determine the chromatin status of these genes in the presence and absence of EKLF. We identified alpha hemoglobin-stabilizing protein (AHSP) by subtractive hybridization and demonstrated a 95 to 99.9% reduction in AHSP mRNA and the absence of AHSP in EKLF-deficient cells. Chromatin at the AHSP promoter from EKLF deficient cells lacked a DNase I hypersensitive site and exhibited histone hypoacetylation across the locus compared to hyperacetylation of wild-type chromatin. Wild-type chromatin demonstrated a peak of EKLF binding over a promoter region CACCC box that differs from the EKLF consensus by a nucleotide. In mobility shift assays, the AHSP promoter CACCC site bound EKLF in a manner comparable to the beta-globin promoter CACCC site, indicating a broader recognition sequence for the EKLF consensus binding site. The AHSP promoter was transactivated by EKLF in K562 cells, which lack EKLF. These results support the hypothesis that EKLF acts as a transcription factor and a chromatin modulator for the AHSP and beta-globin genes and indicate that EKLF may play similar roles for other erythroid genes. PMID- 16705189 TI - Dynamic interactions of chromatin-related mesenchymal modulator, a chromodomain helicase-DNA-binding protein, with promoters in osteoprogenitors. AB - The newly identified protein chromatin-related mesenchymal modulator (CReMM) is expressed by marrow stromal progenitors in vivo and ex vivo. CReMM belongs to a recently identified subgroup of chromodomain helicase-DNA-binding proteins composed of multiple domains including chromodomains, SNF2/ATPase, helicase-C domain, SANT, and A/T-hook-DNA binding domain. Chromatin immunoprecipitation assay was applied to follow the dynamics of CReMM binding to A/T-rich regions on promoters of genes that play a role in osteoblast maturation. CReMM interaction with BMP4 and biglycan promoters in the marrow stromal cells was challenged with transforming growth factor-beta. Treatment with 17beta-estradiol enhanced the binding to estrogen receptor and abolished binding to the prolactin receptor promoters; CReMM interaction with osteocalcin promoter was identified constantly. CReMM binding to the analyzed endogenous promoters suggests its direct role in the transcriptional program activated during osteogenic cell differentiation, which may be a useful tool for following the molecular mechanism of the "stemness" of mesenchymal cells. PMID- 16705190 TI - Endovascular management of unruptured intracranial aneurysms: the dawn of a multidisciplinary treatment paradigm. PMID- 16705191 TI - Unravelling the causes of cerebral damage in subarachnoid haemorrhage: might biomarkers help? PMID- 16705192 TI - Anti-myelin antibodies in multiple sclerosis: clinically useful? PMID- 16705193 TI - Finding silent cerebral aneurysms: the importance of doing nothing. PMID- 16705194 TI - Neurological picture. Diagnosis at the tip of the tongue. PMID- 16705195 TI - Queckenstedt's manoeuvre. PMID- 16705196 TI - Antimyelin antibodies and the risk of relapse in patients with a primary demyelinating event. AB - AIM: To investigate whether the presence of serum antibodies against myelin oligodendrocyte glycoprotein (MOG) and myelin basic protein (MBP) in patients with a clinically isolated syndrome (CIS) predicts the interval to develop more frequently and earlier a first relapse (clinically definite multiple sclerosis: CDMS) than seronegative patients. METHODS: Sera from 45 patients with a CIS and positive intrathecal IgG-synthesis were retrospectively tested for the presence of IgM antibodies against both MOG and MBP. Antibodies were detected by immunoblot using recombinant MOG (1-125) and human MBP antigen preparations. Clinical follow ups were performed retrospectively by telephone interviews and documented neurological examination. RESULTS: Using the Cox proportional hazards model there was no significant increased risk for developing CDMS in anti-MOG and anti-MBP positive patients compared with negative. However regarding the median of the time span between CIS and CDMS over the whole follow up, antibody positive patients (MOG/MBP +/+) developed significantly earlier relapses (median 5.5 months (range 3-20)) than the antibody negative ones (median 25.0 months (range 7 43); p<0.006). On testing sera from 56 apparently healthy students, quite high frequencies of anti-MOG and anti-MBP antibodies (21% and 28% respectively) were detected. This limited specificity of anti-MOG and anti-MBP antibodies has been seen earlier and restricts their diagnostic relevance in MS despite their role as a predictor of relapses after a CIS. CONCLUSIONS: This study confirms previous data only in a subanalysis indicating that patients with positive anti-MOG/MBP antibodies develop earlier relapses than patients who are antibody negative. However, the authors could not verify that the presence of these antibodies anticipates the overall risk of developing CDMS-according to study criteria-after a first demyelinating event within the study period of 21-106 months (mean 60 (SD 25)). PMID- 16705198 TI - Psychosocial impact of finding small aneurysms that are left untreated in patients previously operated on for ruptured aneurysms. AB - OBJECTIVES: In patients with previous subarachnoid haemorrhage (SAH) undergoing follow up screening, the authors assessed the impact of finding but not treating very small aneurysms by comparing quality of life (QOL), anxiety, and depression between patients with a newly detected aneurysm that was left untreated (cases) and patients with a negative screening (controls) as this should be incorporated in the evaluation of effectiveness of screening. METHODS: In patients with previous SAH undergoing screening for new aneurysms the authors compared QOL (SF 36, EURO-QOL, and a screening related questionnaire), anxiety, and depression (Hospital Anxiety and Depression Scale (HADS)) between cases and controls. Differences in scores on the SF-36, EURO-QOL, and HADS were assessed with Student's t test and differences in proportions of patients with HADS scores in the pathological range and screening related changes with chi2 analysis. The authors powered the study to detect a moderate, clinically relevant difference. RESULTS: Thirty five cases and 34 controls were included. Trends for health related QOL, anxiety, depression, and consequences in daily life pointed in the same direction of a less favourable situation for cases but all effects were small, and did not reach statistical significance. On the screenings specific questionnaire, cases more often (but not statistically significant) reported changes in daily life. CONCLUSIONS: The authors found no major or moderate impact on QOL, anxiety, and depression of the awareness of having an untreated aneurysm, which was detected at screening, although most items showed a trend towards more negative effects for cases. Minor effects on individual level cannot be excluded by this study. PMID- 16705197 TI - Axon loss is an important determinant of weakness in multifocal motor neuropathy. AB - BACKGROUND: Multifocal motor neuropathy (MMN) is characterised by asymmetrical weakness and muscle atrophy, in the arms more than the legs, without sensory loss. Despite a beneficial response to treatment with intravenous immunoglobulins (IVIg), weakness is slowly progressive. Histopathological studies in MMN revealed features of demyelination and axon loss. It is unknown to what extent demyelination and axon loss contribute to weakness. Unlike demyelination, axon loss has not been studied systematically in MMN. Aims/ METHODS: To assess the independent determinants of weakness in MMN, 20 patients with MMN on IVIg treatment were investigated. Using a standardised examination in each patient, muscle strength was determined in 10 muscles. In the innervating nerve of each muscle, axon loss was assessed by concentric needle electromyography, and conduction block or demyelinative slowing by motor nerve conduction studies. Multivariate analysis was used to assess independent determinants of weakness. RESULTS: Needle electromyography abnormalities compatible with axon loss were found in 61% of all muscles. Axon loss, and not conduction block or demyelinative slowing, was the most significant independent determinant of weakness in corresponding muscles. Furthermore, axon loss and conduction block were independently associated with each other. CONCLUSION: Axon loss occurs frequently in MMN and pathogenic mechanisms leading to axonal degeneration may play an important role in the outcome of the neurological deficit in patients with MMN. Therapeutic strategies aimed at prevention and reduction of axon loss, such as early initiation of treatment or additional (neuroprotective) agents, should be considered in the treatment of patients with MMN. PMID- 16705199 TI - Axonal damage and outcome in subarachnoid haemorrhage. AB - BACKGROUND: On the basis of preliminary evidence from patients with subarachnoid haemorrhage (SAH), axonal degeneration is thought to be an underestimated pathological feature. METHODS: A longitudinal study in 17 patients with aneurysmal SAH. Ventricular CSF was collected daily for up to 14 days. The neurofilament heavy chain(SMI35) (NfH(SMI35), a biomarker for axonal damage) was quantified using a standard ELISA (upper limit of normal 0.73 ng/ml). The primary outcome measure was the Glasgow Outcome Score (GOS) at 3 months. RESULTS: Of 148 samples from patients with SAH, pathologically high NfH levels in the CSF were found in 78 (52.7%) samples, compared with 20 (5%) of 416 samples from the reference population (p<0.0001). A pathological increase in NfH was observed in all patients with a bad outcome (GOS 1-3) compared with 8% of those with a good outcome (GOS 4-5, p<0.0001). This increase typically became significant 7 days after the haemorrhage (p<0.01). The result was confirmed by analysing the individual mean NfH concentrations in the CSF (3.45 v 0.37 ng/ml, p<0.01), and was reinforced by the inverse correlation of NfH in the CSF with the GOS (r = 0.65, p<0.01). Severity of injury was found to be correlated to NfH(SMI35) levels in the CSF (World Federation of Neurological Surgeons, r = 0.63, p<0.01 and Glasgow Coma Score, r = -0.61, p<0.01). CONCLUSION: Patients with SAH thus have secondary axonal degeneration, which may adversely affect their outcome. PMID- 16705200 TI - Neurological picture. Acute longitudinal myelitis as the initial manifestation of Sjogren's syndrome. PMID- 16705201 TI - Treatment with antidepressants and lithium is associated with increased risk of treatment with antiparkinson drugs: a pharmacoepidemiological study. AB - OBJECTIVE: To estimate the risk for persons treated with antidepressants or lithium of subsequent treatment with antiparkinson drugs (APD). METHODS: The Danish national prescription database supplied data on all persons who received antidepressants, lithium, or antidiabetics (first control group). A second control group was included comprising persons from the general population. Outcome was purchase of APD and the study period was 1995 to 1999. RESULTS: In total, 1 293 789 persons were included. The rate ratio of treatment with APD after treatment with antidepressants was 2.27 (95% CI 2.14 to 2.42) for men and 1.50 (95% CI 1.43 to 1.58) for women. Figures for lithium were almost identical. CONCLUSION: Persons treated with antidepressants or lithium are at increased risk of subsequently treatment with APD, showing an association between anxiety/affective disorder and Parkinson's disease. PMID- 16705202 TI - Epilepsy as a risk factor for cancer. AB - AIM: Epilepsy and long term use of antiepileptic drugs have been suggested to be associated with an increased risk of cancer. The authors therefore set out to analyse previous diagnosis of epilepsy as a risk factor for certain cancer forms in a case control study. METHODS: Incident cases of leukaemia, lymphoma, myeloma, and pancreatic cancer were identified from the Swedish Cancer Registry 1987-99, a total of 52 861 cases. Controls (n = 137 485) were randomly selected from the Swedish Population Registry stratified on age, sex, and year of cancer diagnosis. Cases and controls were linked to the Swedish Hospital Discharge Registry for 1969-99 to identify individuals with first time hospital discharge for epilepsy. RESULTS: While an epilepsy diagnosis the same year as a cancer diagnosis carried an increased risk of non-Hodgkin's lymphoma (OR = 2.89, 95% CI 1.89 to 4.41), Hodgkin's disease (OR = 4.77, 95% CI 1.77 to 13.30), leukaemia (OR = 2.55, 95% CI 1.50 to 4.32), acute myeloid leukaemia (OR = 3.65, 95% CI 1.68 to 7.93), and pancreatic cancer (OR = 2.05, 95% CI 1.22 to 3.45), the authors found no support for an association between discharge with a diagnosis of epilepsy two years or more before the diagnosis of cancer and an increased risk of any of the types of cancer included in this analysis. The lack of association was also evident for individuals with an epilepsy diagnosis preceding malignancy/reference year by >10 years. CONCLUSIONS: Clinical examinations prompted by seizure onset probably mainly explain the observed association between epilepsy diagnoses the year before a cancer diagnosis. However, these results lend no support to the suggestion that epilepsy, and presumably long term exposure to antiepileptic drugs, is associated with an increased risk of the types of cancer included in the present study. PMID- 16705203 TI - Positional nystagmus and vertigo due to a solitary brachium conjunctivum plaque. AB - The authors describe two patients suffering from demyelinating central nervous system disease who developed intense vertigo and downbeat nystagmus upon tilting their heads relative to gravity. Brain MRI revealed in both cases a single, small active lesion in the right brachium conjunctivum. The disruption of otolithic signals carried in brachium conjunctivum fibres connecting the fastigial nucleus with the vestibular nuclei is thought to be causatively involved, in agreement with a recently formulated model simulating central positional nystagmus. Insufficient otolithic information results in erroneous adjustment of the Listing's plane in off-vertical head positions, thus producing nystagmic eye movements. PMID- 16705204 TI - Incremental limb hypometria. AB - To find out if patients with right hemisphere strokes (RHD) demonstrate a form of progressive or incremental limb hypometria (ILH), 11 RHD patients and eight matched controls were tested by having them draw a series of horizontal lines while blindfolded. Unlike controls, six RHD patients displayed an incremental decrease in the length of lines they drew, a sign of ILH. ILH might be a common source of disability and hinder rehabilitation efforts. PMID- 16705205 TI - Increased extrapyramidal symptoms in patients with schizophrenia and a comorbid substance use disorder. AB - Few data have been gathered about the impact of psychoactive substances on extrapyramidal symptoms (EPS) in schizophrenia, and so far, inconsistent results have been reported. We studied 41 outpatients with schizophrenia (based on DSM-IV criteria), who were divided into two groups: with (n = 17) and without (n = 24) a substance use disorder (alcohol, cannabis, and/or cocaine). Both groups were matched for sociodemographic data and psychiatric symptoms (Positive and Negative Syndrome Scale). EPS were evaluated with the Extrapyramidal Symptoms Rating Scale and the Barnes Akathisia Scale, and all patients were stable on either quetiapine or clozapine. Patients receiving anticholinergic drugs were excluded. Analyses of variance were conducted on both groups and showed that schizophrenia patients with a comorbid substance use disorder (especially cocaine) displayed more EPS compared with non-abusing patients. PMID- 16705206 TI - Leech therapy in the treatment of median nerve compression due to forearm haematoma. PMID- 16705207 TI - Misleading conclusions on rt-PA treatment in the very elderly. PMID- 16705208 TI - Remission of chronic inflammatory demyelinating polyneuropathy after alemtuzumab (Campath 1H). PMID- 16705209 TI - Chronic intestinal pseudo-obstruction as the initial feature of pure autonomic failure. PMID- 16705210 TI - Ear dyskinesia. PMID- 16705211 TI - Simultaneous occurrence of neuromyotonia and morphoea: a cause-effect relationship? PMID- 16705212 TI - Thematic review series: patient-oriented research. Imaging atherosclerosis: state of the art. AB - The ability to image obstructive arterial disease brought about a revolution in clinical cardiovascular care; the development of newer technologies that image arterial wall thicknesses, areas, volumes, and composition allows valid imaging of atherosclerosis for the first time. Development of noninvasive imaging of atherosclerosis has further led to a quantum shift in research in the field by enabling the study of asymptomatic populations and thus allowing investigators to focus on preclinical disease without the many biases associated with the study of symptomatic patients. These noninvasive investigations have broad implications for clinical care as well. Coronary angiography, computed tomographic (CT) imaging of coronary calcium, intravascular ultrasound, multidetector CT angiography, B mode ultrasound of the carotid arteries, and MRI of the carotid arteries all have unique strengths and weaknesses for imaging atherosclerosis. Certain of these techniques are extremely useful as outcome variables for clinical trials, and others are uniquely useful as predictors of the risk of cardiovascular disease. All are informative in one way or another with regard to the role of plaque remodeling and composition in disease causation. CT and MRI technology are advancing very rapidly, and research and clinical uses of these imaging modalities promise to further advance our understanding of atherosclerosis and its prevention. PMID- 16705213 TI - Quantitative analysis of SR-BI-dependent HDL retroendocytosis in hepatocytes and fibroblasts. AB - Previous studies have suggested that HDL retroendocytosis may play a role in scavenger receptor class B type I (SR-BI)-dependent selective lipid uptake in a cell-specific manner. To investigate this possibility, we developed methods to quantitatively measure HDL uptake and resecretion in fibroblast (COS-7) and hepatocyte (HepG2) cells expressing exogenous SR-BI. Approximately 17% and 24% of HDL associated in an SR-BI-dependent manner with COS-7 and HepG2 cells, respectively, accumulates intracellularly after a 10 min incubation. To determine whether this intracellular HDL undergoes retroendocytosis, we developed a pulse chase assay whereby internalized biotinylated (125)I-HDL(3) secreted from cells is quantitatively precipitated from cell supernatants using immobilized streptavidin. Our results show a rapid secretion of a portion of intracellular HDL from both cell types (representing 4-7% of the total cell-associated HDL) that is almost complete within 30 min (half-life approximately 10 min). In COS-7 cells, the calculated rate of HDL secretion ( approximately 0.5 ng HDL/mg/min) was >30-fold slower than the rate of SR-BI-dependent selective cholesteryl ester (CE) uptake ( approximately 17 ng HDL/mg/min), whereas the rate of release of HDL from the cell surface ( approximately 19 ng HDL/mg/min) was similar to the rate of selective CE uptake. Notably, the rate of SR-BI-dependent HDL resecretion in COS-7 and HepG2 cells was similar. BLT1, a compound that inhibits selective CE uptake, does not alter the amount of SR-BI-mediated HDL retroendocytosis in COS-7 cells. From these data, we conclude that HDL retroendocytosis in COS-7 and HepG2 cells is similar and that the vast majority of SR-BI-dependent selective uptake occurs at the cell surface in both cell types. PMID- 16705214 TI - [Liver resection with regional chemotherapy in the cases of colorectal metastasis in the liver]. AB - The surgical procedure is the most important in the combined treatment of the liver colorectal metastatic injury. The aim of our investigation was to determine the efficacy of liver resection together with regional chemotherapy(R/C) at solitary injury of liver. 48 patients with metastatic damage of the liver from the colorectal cancer were observed. Different degree of liver resection with R/C was performed. Outcomes of partial resection of the liver metastasis combined with primary tumor resection and 5-FU chemotherapy in the observed cases were quit satisfactory (5 year survival-14,6%). PMID- 16705215 TI - Prostate cancer detection rate in patients with obstructive voiding symptoms by sextant biopsy: preliminary results. AB - To evaluate the effectiveness of a laterally directed sextant biopsy in the group of high volume prostate patients with obstructive voiding symptoms and suspected of prostate cancer. Biopsy was performed in 197 men (age: median 63 years, range 47 to 82 years) because of elevated PSA and/or abnormality in digital rectal examinations (DRE). In most cases, a markedly enlarged prostate was noted: the mean prostate volume was 79,9+/-38,5 cc, and in 73% of the cases, the volume of the prostate was >50 cc. Prostate cancer was diagnosed in 56 of 197 cases (28,4%). The prostate cancer detection rate among patients with a prostate volume of 80cc (high volume) were 39,6%, 32,8% and 18,6%, respectively. Our results showed that the laterally directed sextant biopsy with the overall detection rate as high as 28,4% and very low complications is an effective method for the detection of prostate cancer among the suspected patients with obstructive voiding symptoms and markedly enlarged prostates. PMID- 16705216 TI - [Treatment of abdominal lymphosarcoma in children]. AB - Gastrointestinal tract and abdominal lymph nodes are common sites for development of lymphosarcomas in children (36-53% of all lymphomas). Lymphosarcoma is a malignant tumor developed from the lymphoid tissue and characterized with broad spectrum of clinical manifestations, inhomogeneous course of the disease and prognosis. 25 (24,0% of all abdominal lymphosarcomas) patients with intestinal lymphosarcoma admitted to the childhood tumors department of the National Cancer Center of Georgia from 1980 to 2006. One of the most informative diagnostic methods for this disease is X-ray examination. Ultrasound and CT are leading investigating methods giving us possibility to determine topography of the tumour as well as to evaluate efficiency of the specific treatment and carry out a dynamic control of the patients after the complete remission. In all patients with intestinal lymphosarcomas surgical treatment without any preoperative specific therapy (chemotherapy, radiotherapy) has been carried out. Analysis of the specific treatment results obtained has shown that intestinal lymphosarcoma is the disease with worse prognosis in comparison with abdominal lymph nodes lymphosarcoma. It has been ascertained that in 88,0% of abdominal lymph node damages complete cure can be achieved using surgery and rational program of ACOP chemotherapy. PMID- 16705217 TI - [Influence of clinical features of oral cancer on incidence of regional metastasis]. AB - The regional metastasis has a high frequency in oral cancer and significantly influences the prognosis of this disease. It is known that some clinical features determine the incidence of regional metastasis. Accordingly, the study of these features represents an important issue in the improving the results of the treatment. The aim of our study was the evaluation of different clinical features in relation to the regional metastasis. The study group was composed by 478 patients. It was revealed that the patient's sex, age, period of tumor existence, tumor ulceration and the pre-cancer diseases doesn't influence the incidence of regional involvement. It was found that the primary tumor sizes, the pattern of growth, invasion in surrounding tissues, number of involved anatomical regions and the pain symptom are significant clinical features of regional metastasis. The statistical evaluation of received results, along with the consideration of pathology criteria will allow (in the framework of future study) to work out of individual prognosing method for development of regional metastasis in oral cancer. PMID- 16705218 TI - [Possibilities of colposcopy in the diagnostics of cervical Chlamydia mycoplasmatic infections (population prospective study)]. AB - Possibilities of colposcopy in the diagnostics of cervical chlamydia-mycoplastic infections are less investigated. The aim of given work was the study of this problem. Colposcopic examination was performed in 952 women (age - from 16 to 67) asked to clinics for examination and more precise definition of diagnosis because of different complaints. It was revealed specific colposcopic features of chlamydia-mycoplastic genital infections, which were compared with the results obtained from "gold standard" methods - direct immunofluorescence method and bacteriological analysis of cervical smears. High sensitivity (Se) - 0,96, specificity (Sp) - 0,79 and diagnostic effectiveness (De) - 0,93 of colposcopy were established. Authors conclude that the application of colposcopy for diagnostics of chlamydia-mycoplastic genital infection is particularly important: during urgent cases; during the case, when it is impossible to carry out the deep diagnostics of the sexually transmitted infections; for optimization of the diagnostics of the sexually transmitted infections. PMID- 16705219 TI - [Some psychological characteristics of pregnant teenagers in the first trimester of pregnancy]. AB - The purpose of the study was the investigation of emotional sphere of pregnant teenagers in the third trimester of pregnancy. The results of the trial demonstrate: 1) from the point of view of emotional sphere, pregnant teenagers, despite of the physiological course of the pregnancy, represent the group of risk, but the sphere of teenagers with pathological course of the pregnancy is more interesting. 2) in the group of pregnant teenagers with pathological course of the pregnancy considerably higher was the number of women who showed different types of fear, which as a matter of fact in intensity differed from fears of pregnant teenagers with physiological course of the pregnancy. 3) according to the data of the study anxiety is significantly expressed among teenagers with pathological course of the pregnancy. PMID- 16705220 TI - [Induced hypothermia during low-flow anesthesia in elderly patients]. AB - The purpose of the study was to investigate the possibility of reducing of level of induced hypothermia during general balanced anesthesia, by using of low - flow techniques of anesthesia in elderly and senile patients. The temperature homeostasis was studied intraoperatively in 76 patients, aged 65-84 years, ASA III - IV, underwent different surgical operations under high (n=34) and low - flow (n=42) anesthesia. It was demonstrated that the low - flow anesthesia allows not only reduce loss of moisture and heat from the patients airway during surgery, but also optimize the intraoperative patient - body temperature, decrease level of induced hypothermia and minimize frequency of complication due to temperature misbalance in the immediate postoperative period. Such anesthesia is a sufficiently effective and safe method of inhalation anesthesia, which can be used with good results in routine geriatric surgery. PMID- 16705221 TI - [Treatment of recurrent aphthous stomatitis with application of a preparation of a hesperidin 5.0%]. AB - Open randomized 2-year trial has been performed to analyze effectiveness of Unguentum of a hesperidin developed by us for the treatment of recurrent aphthous stomatitis. 80 patients with recurrent aphthous stomatitis in the age from 15 till 60 years from the contingent of the polyclinic of the veterans of Tbilisi were studied. After inclusion of the preparation into the treatment the pain and duration of inflammatory process have been decreased; the complications were not observed. In the group of patients getting treatment with hesperidin, frequency of unsatisfactory outcomes decreased significantly (chi(2)=9,8, p < 0,05). Reduction of relative risk of unsatisfactory outcomes was about 0,8 (95% CI: 0,29 0,86), reduction of absolute risk - 0,33 (95% CI: 0,13-0,52). It may be concluded that application of a preparation may be considered as one of the options for the treatment of recurrent aphthous stomatitis. PMID- 16705222 TI - [Influence of whitening endoopalescence systems on the restoration of devitalized teeth]. AB - At the end 80s of the XX century bleaching of teeth became one of the most important issues in the stomatological industry. Currently there are two methods of bleaching: external and internal (endo bleaching) methods. It is proved that the adhesive ability of the tooth tissues reduces as a result of peroxide penetration into enamel and dentine. Based on this, the aim of our study was to use the antioxidants for excluding the peroxides from tooth tissues and for rehabilitation of reduced adhesion. For this purpose we used central and side 36 healthy incisors, where after processing of tooth tissue with antioxidants and without them we studied the formation of micro rift between the operation material and dentine, and intensity of micro flowing by means of stereo microscope. We used 30% endo-opalescence as a bleacher and used 10% sodium acrobat as antioxidant. As a result of experiment we found out that the teeth where we used antioxidant were not penetrated with metaline blue; unlikely, the teeth where we did not use the same method we got the approximate result in the group of teeth, where for peroxide exclusion we performed the crown reconstruction not immediately, but after 7 days. During the whole experiment the teeth were kept in thermostat at 37 degrees C in saliva to create the natural conditions for them. In overall we can conclude that the deteriorated adhesive ability with peroxide influence can be improved by application of antioxidants. PMID- 16705223 TI - [Histological characteristics of stimulating role of hydroxyapatite and platelet rich plasma in reparative processes in experiment]. AB - Calcium and Phosphorus are the basic elements in making the inorganic matrix of bone tissue. With the purpose of our research was to study the local effect of hydoxypatite and Platelet Rich Plasma (P.R.P.) on the course of reparative osteogenesis. We used hydroxyapatite granules made by the firm "Polystom" with the chemical formula Ca(10)(PO(4))(6)(OH)(2). Platelet Rich Plasma we have got from blood of rabbits by centrifuging. Experiment was made on 30 rabbits. by operation the standard defects were created with the diameter of 66mm and the depth of 4-5mm on the mandibular bone of the rabbits. In the laboratory group defects were filled with hydroxyapatite and P.R.P., in control group with their own blood. After 7,14,21,35 and 50 days after operation three rabbits were taken from each group. Mandibular bone fractions with defects were relieved from soft tissue, were fixed in 10% neutral formalin and were stained with hematoxyn eozine. The study showed that in the study group at the initial stage of differentiation from the moment of traumatization the pre-osteoblast and osteoblast contour permanently appears. And in the control group the resorbtion processes were clearly expressed. At the end of the experiment of 50 days in the experimental group the compaction of osteogenic structure was observed in all defect areas. Newly generated tissue underwent compaction. The results of the experiment prove that the hydroxypatite and Platelet Rich Plasma has a stimulative role in the process of osteogenesis. Activation of the reparative processes is far more intensive at the final stage of osteogenesis. PMID- 16705224 TI - [Heart arrhythmias and congestive heart failure]. AB - Along with the successful pharmacotherapic achievements in patients with congestive heart failure survival rate in this category of patients still remains unfavourable. Many antiarrhythmic drugs that cause proarrhythmic effect in patients with congestive heart failure, are characterized by narrow therapeutic window of action and their metabolism is usually changed. Patients with ventricular arrhythmias and disturbances of left ventricular function belong to group with elevated risk of sudden death and complex ventricular impairments are predictors of lethal outcome. Use of ACE inhibitors, angiotensin II receptor inhibitors, beta-blockers (in order to control ventricular rhythm during tachycardia) in combination with antiarrythmic drugs increases effectiveness and safety of therapy, lowers degree of congestive heart failure development and improves prognosis of diseases of that kind. Utilization of nonarrhythmic drugs during treatment of arrhythmias including atrial fibrillation, is novel approach in the treatment with antiarhythmic drugs. PMID- 16705225 TI - Peculiarities of lipid profile parameters in cardiac patients with hypo- and hyperthyroidism. AB - The problem of thyroid dysfunction is very actual in cardiac patients. The aim of the present study was to evaluate lipid profile parameters in 73 hypo-, hyper- and euthyroid cardiology patients visited the Emergency Cardiology Center. Most of the patients had coronary artery disease. Thyroid dysfunction was diagnosed on the basis of patients' clinical status and blood thyroid stimulating hormone. In hypothyroid patients blood levels of total cholesterol, low-density lipoprotein cholesterol and triglycerides were increased. In contrast, the patients with hyperthyroidism had normal concentrations of these parameters. Exception was antiatherogenic fraction of lipid profile - high-density lipoprotein cholesterol, which mean levels were normal in all patients' groups. It is noteworthy that the highest levels of atherogenic fractions were observed in the groups of patients who had hypothyroidism in combination with diabetes mellitus. In conclusion, the results of present study showed that thyroid dysfunction may play a potential independent role in changes of atherogenic lipid profile. PMID- 16705226 TI - [Some peculiarities of the activity of postheparin lipoprotein lipase and parameters of lipid metabolism at ischaemic heart disease]. AB - The study of the relation of atherosclerosis with lipid metabolism (LM) remains still very important. The aim of presented work was the investigation of peculiarities of lipoprotein lipase (LPL) and LM at IHD in different age and sex groups; also the search of possible relation of LPL activity changes with myocardial contractility function and coronarography data at IHD. 47 patients (15 females, 32 males) with IHD were investigated. They were divided on 3 groups: 1 <50 years (n=14); 2 - 50-59 years (n=14); 3 - >/=60 years (n=19). Patients were investigated by coronarography and echocardiography. Main parameters of LM were determined by enzyme-linked immunoassay. LPL activity was defined by potentiometric method. It was observed significant difference between levels of LPL activity of different age groups (p(1-2)=0,027, p(1-3)<0,001, p(2-3)=0,035). Significant difference in this parameter by sex feature did not observed. The degree of damage in anterior descending branch of left coronary artery negatively correlated with LPL activity (r=-0,5598, p<0,001). Ejection fraction (EF) revealed positive correlation with LPL activity (r=0,4362, p=0,002). The degree of damage of left coronary artery, in particular, medial segment of circumflex branch correlated with LM parameters. EF positively correlated with high-density lipoprotein cholesterol blood level (r=0,3548, p=0,036). LPL activity in patients with IHD significantly decreased with aging. From obtained results it will be supposed that LPL activity and LM parameters influences on the damage degree, basically, left coronary artery. PMID- 16705227 TI - Education and coronary heart disease. AB - In Georgia, like the other post soviet republics CHD morbidity is increasing, especially among young and middle aged people i.e. among those who should have the most working ability that points at both individual and social significance of the problem. CHD is becoming more and more common among rural inhabitants, different professional groups involved not only in mental but also in physical work. There is growing evidence that socio-economic status (SES) is one of predictors of coronary heart disease and its risk-factors, though the direction of the relationship is not stable and differs by countries. SES is characterized by factors as education, occupation, income, housing conditions, etc. Education is the most widely used measure of socio-economic status in epidemiologic studies. Our research aimed assessment of the relationship between education and coronary heart disease that will contribute to selection of target groups and correct planning of primary and secondary prevention measures. A case-control study was carried out. The study results indicate that Georgia is in the transitional period and probably in several years education and CHD will have negative correlation. PMID- 16705228 TI - [Occurrence of pathologies of the psychic sphere among the patients in the period of the angioplasty on coronary blood vessels]. AB - The aim of our study was to reveal pathologies of the psychic sphere (symptomatology of alarm disorders, depression and sleep disturbance) in several days after the angioplasty on coronary blood vessels. There have been examined 70 patients (20 women and 50 men) at 3-5 days after the angioplasty on coronary blood vessels. Their age ranged from 40 to 75 years. The main cause of hospitalization of these patients was the acute coronary syndrome in 50% of the cases, non-stable stenocardia in 20% and acute myocardial infarction on 30% of the cases. There has been studied the percentage rates of alarm disorders, depression and sleep disturbance among these patients. It was revealed that alarm disorders in high level occurred in 54,28%, depression in 32,85% and sleep disturbance in 72,85%. Among the majority of these patients the degree of coronary occlusion was from 70 to 90%. On the basis of the above-mentioned indices there can be concluded that there exists a considerable correlation between the degree of coronary occlusion and the occurrence of pathologies of the psychic sphere. PMID- 16705229 TI - Influence of nebivolol on endothelial dysfunction in salt-sensitive hypertension. AB - The study was aimed to assess influence of nebivolol (nebilet) on endothelial factors (NO and endothelin1) during treatment of salt-sensitive hypertensive patients. A total of 40 patients (18 females and 22 males, mean age 53,1+/-1,58 yrs) with mild (I stage) arterial hypertension (with diastolic pressure 90-99 mm Hg and/or systolic pressure 140-159 mm Hg) were studied. Twelve age matched healthy subjects (4 women and 8 males) composed control group. Salt sensitivity was detected by M. Weir (1993) method. Patients were divided in two groups: salt sensitive hypertension (group I, 16 patients) and salt-resistant hypertension (group II, 24 patients). Nebivolol (Nebilet, Berlin-Chemie, Germany) was administered to 25 patients in dosage of 5 mg once a day during two weeks. Our studies have showed that ET-1 plasma levels were significantly elevated in patients with essential hypertension, while NO was markedly reduced compared to control group. NO levels were slightly lower in salt-sensitive hypertensives than in salt-resistant patients (p>0,05). ET-1 levels expressed opposite changes: in patients with salt-sensitive hypertension ET-1 levels almost twice exceeded those of salt-resistant patients (p<0,01). After treatment with nebivolol NO plasma level in salt-sensitive hypertensives significantly increased in Group I and did not change significantly in salt-resistant patients. Only slight decrease in ET-1 levels was detected in both groups after taking nebilet. Nebilet (nebivolol) treatment has an influence only on NO levels and appears to be the most effective in salt-sensitive hypertension. PMID- 16705230 TI - [Activity of antioxidative ferments during obesity of different degree]. AB - The purpose of our research was to define influence of dislipidemia on the redox system, developed during obesity. 150 patients from 18 to 55 years were examined having obesity of a different degrees who have been admitted to the endocrinology department of 4-th clinical hospital of Tbilisi. On the basis of the analysis of the received data it is possible to conclude that with in a degree of obesity a deterioration of parameters of lipid exchange was observed which from its part exerts its influence on the redox-system of an organism. Namely, it promotes reduction of antioxidant protection system in comparison with the control group. Also, dependence of change of activity of antioxidant ferments on the age and intensity of lipid exchange has been revealed. Furthermore, it has been revealed that deterioration of lipid exchange in patients over 30 years is the reason for compensatory activity of antioxidant ferments (SOD and catalase). PMID- 16705231 TI - [The effect of diet and exercise on the parameters of lipid metabolism in the patients with metabolic syndrome and impaired carbohydrate metabolism of various degrees]. AB - The aim of our study was the investigation of the peculiarities of lipid metabolism in the patients with metabolic syndrome and impaired carbohydrate metabolism of various degrees after diet and moderate exercise. We selected 123 overweight patients from Imereti Region. Metabolic syndrome in patients was determined by World Health Organization -2002 classification. Total cholesterol, triglycerides, high density lipoprotein, low density lipoprotein, very low density lipoprotein and atherogenic indices have been measured. Diet and exercise - changes in the life style decreases the risk of metabolic syndrome development and decrease fasting and postprandial glucose. It was also indicated that the parameters of lipid metabolism sharply improved by diet and exercise and the degree of improvement was practically identical in patients with diabetes mellitus type 2 and impaired glucose tolerance. PMID- 16705232 TI - Study of chemical composition of glue "RAZI" used by solvent abusers in Tbilisi. AB - The analysis of the glue "RAZI" was carried out at the Republican Chromatographic Center of Georgia, with high-efficient liquid chromatographic equipment "Millipore Waters" (USA), by I. Wagner's Method. We were able to determine presumable composition of the glue chemical components. The different substances were evaporated at different times fixed by appropriate peaks on chromatogram. There have been at least five toxic substances identified within the glue composition, which are used as industrial or household chemical goods and as some authors explain, may be used as inhaling psychoactive substances. These substances are: benzol (benzene), toluene (toluol), phenol (carbolic acid), chloroform, and methyl-ketone. The glue is inhaled with a small polyethylene bag. The substances are evaporated during inhalation and join arterial blood through lungs. In several minutes leap forward a situation similar to alcohol intoxication. The evaporated substances characterized by strong toxicity and influence to the organism in different ways. The results of chronic glue abuse is serious, such as lethal aplastic anemia, leukemia, marrow damage, chromosome aberrations, functional disorders of CNS, dystrophic changes in myocardium, cirrhosis, liver atrophy and so on. Abuse of the glue "RAZI" can lead to very serious medical consequences and represents emerging public health problem in Georgia. PMID- 16705233 TI - Left ventricular morphological changes due to vigorous physical activity in highly trained football players and wrestlers: relationship with aerobic capacity. AB - The heart undergoes profound changes in response to systematic athletic training. Several adaptations of cardiac shape and function occur with athletic training to improve the heart's function as a pump and thereby increase aerobic capacity. Maximal oxygen consumption or VO2 max is regularly used as an index of physical fitness but the issue of which of the left ventricular structural parameter correlates with VO2 max remained unresolved. The aim of the study was to examine the effects of the long-term intensive physical training on cardiac responses in highly trained athletes-football players and wrestlers- and reveal the structural parameter of the heart which better correlates with aerobic capacity. We studied a group of highly trained male athletes, 221 football players and 51 wrestlers, and 48 healthy male sedentary controls. The research has included M and 2D echocardiography, resting ECG and stress-testing. The data indicate that highly trained male athletes had higher value of maximal oxygen uptake PWC(170) and PWC(170)/kg than untrained male controls; they exhibit greater left ventricular internal dimension, left ventricular wall thickness, relative wall thickness, left ventricular mass and mass index compared to the untrained controls. The amount of physiologic hypertrophy that occurs in the athletes is related to the intensity and duration of the exercise and is directly related to the fitness level or V02 max. It was concluded that VO2max is the variable that better correlates with the LVMI. PMID- 16705234 TI - Lower extremity vein digital photoplethysmography in highly qualified football players and wrestlers. AB - Modern sport, along with the high technical, tactic and psychological readiness, requires good physical preparation achieved by a big physical load during trainings and competitions. Aim of the investigations was to study lower extremity venous system functional condition during physical load in highly qualified football players and wrestlers. Highly qualified 25 football players and 30 wrestlers, of age 18-25 years, were studied. Olympic, World and Europe champions were included among wrestlers. Lower extremity venous system digital photopletismography (D-PPG) was conducted by apparatus: Rheo Dopplex II of Huntleight Diagnostics. The equipment digitally analyses photopletismoghraphic plots. Significant functional parameters: vein restoration time (VRT) and venous pump function (VP) in deep venous system were reported by the apparatus appropriate software. It included parameter interpretation diagram, defining degree of venous system condition as normal, equivocal or pathological. Target sportsmen were examined in siting position with physical loading pedis. Sportsmen lower extremity vein light transducing change registration associated with the degree of vessel filling was studied by digital photopletismography . The study results showed that vein restoration time (VRT) in football players is two times minor to compare to the one of wrestlers, while venous pump (VP) function is about 1.5 times higher in footballers than in wrestlers. Degree of lower extremity venous system condition according to parameter interpretation diagram program showed normal values only in few cases. Slight deviations were observed only in individuals with recent trauma. Further examination after the appropriate treatment revealed normal values in traumatic subjects. The search also showed that digital photopletismography appears to be effective, simple and financialy available diagnostic method, which should be more frequently applied in lower extremity vein function examinations among sportmen of different kinds as well as in estimation of recovery therapy effectiveness in post-trauma period. PMID- 16705235 TI - [Peculiarities of joint course of allergic rhinitis and bronchial asthma in population of school-age children]. AB - Purpose of the work was to investigate clinical-allergic peculiarities of allergic rhinitis, together with bronchial asthma in school-age children. There were studied random representative group of children, in total 3000 subjects from 6 to 17. According to the results of epidemiological research in school-age children's population, prevalence of allergic rhinitis was 13,2%. In 19,1% of cases, allergic rhinitis was accompanied with bronchial asthma and in 8% of the patients there was indicated bronchial spasm of physical load (verified as clinically, also through the test with physical load), coughing version of asthma was observed in 4,5% of children. Thus, in children with allergic rhinitis, within the structure of clinical versions (by the criteria of severity) of rare bronchial asthma prevailed (69,8%, among which 65% - intermitting form and 4,8% - persisting form), without statistically significant difference between the boys and girls. Combination of inherited predisposition to allergy with the atopic dermatitis, in the in the anamnesis of children with the increased level of general IgE in the blood serum may be considered as early diagnostic predictor of atopic phenotype as for the allergic rhinitis, also for its combination with the bronchial asthma. PMID- 16705236 TI - [Role of oxygen-nitrogen stress in neonatal sepsis]. AB - Reference data indicate on the important role of free radicals of nitrogen and oxygen in the mechanisms of the damage of cells and tissues during neonatal sepsis. Meanwhile these connections play an important role in protecting of the host organism from intervention of microorganisms. The aim of the study was the evaluation of peculiarities of oxidizing metabolism and containing of nitrogen oxide. Low consistence of nitrogen oxide and the disturbance of the intracellular oxidative-resorting balance during neonatal sepsis, especially in premature children, induces insufficiency of immune and antioxidant system, which changes the homeostasis and develops the oxidative stress of the host organism. As it was shown by the results of our study, antibacterial and supportive therapy doesn't provide the important reduction of processes of free radical oxidizing. The last circumstance makes a barrier for improving a clinical condition of the newborns. PMID- 16705237 TI - [Clinical characteristics of juvenile systemic hypertension in the population of Tbilisi]. AB - The goal of the work was to study a prevalence of juvenile systemic hypertension among 15-16 years old adolescents of Tbilisi, its clinical characteristics and several indices of mental capacity. In the result of an analysis of 378 adolescents' blood pressure records only 6 (1.6%) had high normal values and 32 (8,46%) revealed juvenile systemic hypertension. Four youths from 6 with high normal values refused to participate in a further investigation. For this reason the study was continued only in 32 youths with juvenile systemic hypertension. In the contingent noted 6 (18,75%) had overweight, 9 (28,12%) had hereditary predisposition, 15 (46,9%) revealed excessive use of salt, 3 (9,4%) had moderate consumption of alcohol and 7 (21,9%) noted insufficient physical activity. Juvenile systemic hypertension in youth was predominantly asymptomatic, only 9 (28,12%) had mild headache in the evening, 12 (37,5%) had light fatigue and 5 (15,6%) weak dizziness. Fourteen youths noted diminished work-status and being behind in learning process. In the persons with juvenile systemic hypertension diminution of the attention concentration was observed during a fulfilment of a simple sensor and motor task in condition of monotonous work (increment of answers missed up to 13+/-12%). A psycho-emotional loading test has proved the fact of mental capacity lowering within the persons with juvenile systemic hypertension that is expressed in the increment of errors made up to 12+/-3,1% and answers missed up to 11+/-2,3%. Our investigations confirm the reasonability of blood pressure monitoring among adolescents for the early detection of the juvenile systemic hypertension and the conduction of an opportune preventive measures for avoiding of cardiovascular diseases in a mature age. PMID- 16705238 TI - [Biochemical and clinical aspects of melatonin biosynthesis induced by noradrenalin in experiment]. AB - Melatonin has been considered as a hormone with potent immunomodulating, anxiolitic and antioxidative properties. Melatonin is also known to be inhibitor of sympatoadrenal activity. The study was aimed to elucidate changes in melatonin synthesis induced by noradrenalin IM injection. Melatonin plasma levels were detected by radioimmunoassay. All studies were performed conducted on white non linear rats. By the 20th day after noradrenalin administration plasma level of melatonin was significantly decreased. In conclusion, short-term activation of sympatoadrenal system (i.e. stress-induced activation) is accompanied by elevated plasma levels of melatonin, however long-term hyperactivity of the sympatoadrenal system atrophy of epiphiseal parenchyma develops followed by decreased melatonin synthesis. It has been considered that melatonin might contribute to chronic stress-induced process associated with involution of thymus and lymphatic nodes, hypertrophy of adrenal cortex. Data obtained might be taken into consideration during the treatment of stress-induced conditions. PMID- 16705239 TI - [Influence of some low doses of ionizing radiation on the transmembrane transport of sodium, potassium and calcium ions in normal and tumor cells in experiment]. AB - Problem of treatment of various malignant tumors is one of the most acute ones in contemporary medicine. In evaluation of treatment efficiency it is important to implement the sensitive tests, which allow precise determination of treatment agent, specifically that of ionizing irradiation. One of such sensitive indices is the rate of transmembrane transport of Na(+), K(+), and Ca(2+), as well as enzymes controlling this transport. In the present work characteristics of transport of these ions and enzymes were assessed during low dose (0,05 and 0,1 Gy) irradiation, which attracts increased attention nowadays. Recording of the ions' concentration was performed with the ion-selective electrodes. The cells of Erlich carcinoma were studied. It was found that irradiation with doses of 0,05 and 0,1 Gy, unlike dose of 0,5 Gy, exerted a stimulating effect on the ion transport. The same is true in regard of enzymes, which control transport of ions, especially those involved in Ca(2+) transport. The mechanism of this effect is unclear as yet. PMID- 16705240 TI - [Functional state of the dopaminergic system of the brain on a background of different doses of amitriptiline]. AB - The aim of the work is study of adsorbtion dynamics of amitriptiline on the surface of erythrocytes and to compare the obtained data with the character of pharmacokinetics and pharmacodynamics of amitriptiline in plasma. With the method of PLC the adsorb gas was been analyzed. The drag was administered to adult drugs and the blood sampled were collected following 10, 20, 60 min. and 4,0; 6,0 and 8,0 hours after the injection. In the supernatants from the erythrocytes and in the blood plasma the dopaminergic systems has been analyzed. Our data proved the ability of amitriptiline to adsorb on the surface of erythrocytes. Its concentration in the supernatant washed from the surface of erythrocytes and in the blood were not the same. It may be connected with the character and quantity of functional groups located on the surface of erythrocytes. PMID- 16705241 TI - [Dissolution study of the gentin tablets]. AB - To establish individual dissolution test for the tablets gentin, which have a marked influence on the secretory function of the stomach (increase the free acidity of the gastric juice and stimulate enzymoproduction function of the stomach), and gastroprotective functioning, the conditions for the test realization were studied and specified, using apparatus "Rotating Basket" (Dissolution test, apparatus 1, p.1791, USP XXIII, 1995). For the quantification of the active ingredients in buffer solutions chromatospectrophotometric method was used. The same conditions were used for the accomplishment of the dissolution test using apparatus "Rotating Paddle" (Dissolution test, apparatus 2, p.1791, USP XXIII, 1995). Metrologic characteristics were compared for the evaluation of the reproducibility of the results. Obtained data showed better reproducibility of the results for the apparatus "Rotating Paddle" compare to the apparatus "Rotating Basket". Estimated conditions are useful for the routine control of the tablets "Gentin" in the case of manufacturing. PMID- 16705242 TI - Prognostic significance of clinical and immunological parameters in diffuse large B-cell lymphomas. AB - The aim of the study was to evaluate influence of CD10 expression on overall survival with risk factors according of the IPI. CD10 expression was evaluated in 45 patients with diagnosis of Diffuse large B-cell lymphomas. Unfavorable parameters of the International prognostic index (IPI) were compared in the CD10 positive and CD10 negative groups, number of unfavorable clinical findings in CD10 positive and CD10 negative cases were similar at diagnosis. The study revealed that the CD10 antigen expression is an unfavorable prognostic factor in the high and high-intermediate risk groups regarding to the IPI, overall survival is shorter in the CD10 positive cases and was 14,1 month compared with 23,5 month in CD10 negative group(p<0,01). CD10 antigen expression may have importance as one of the prognostic factors in patients with diagnosis of diffuse large B-cell lymphomas. Interestingly, that influence of CD10 expression on overall survival in the low and low-intermediate risk groups (IPI) wasn't revealed (p>0,05). PMID- 16705243 TI - Influence of toluene intoxication on the number of mitral and granular neurons in olfactory bulbs of rats. AB - In the last years, implementation of the volatile solvents in order to get a narcotic high, used via inhalation (sniffing), increased significantly, especially in the youth. Substances containing these agents induce hallucinations and euphoric state. In the present study the influences of toluene intoxication on quantity of the mitral and granular cells in olfactory bulbs was analyzed in young and adult rats. An inhalation route of intoxication was used in our experiments - animal inhaled toluene until attaining a sidewise laying position. This procedure was performed in the closed glass container, in which air was beforehand saturated with the toluene vapours. Experiments was carried out on two age groups (n=20 animals): one - and two months age rats. Inhalation period was 40 days. Each age group consisted of ten animals and was divided into the two subgroups: I - control animals (n=5), II - experimental animals (n=5). Assessment of the neurons' quantity in different structures was made according to the fractional approach method proposed by M. West (1999). The results obtained showed that number of mitral neurons in the olfactory bulb of the Subgroup II of the rats, sniffing since the age of one and two months, significantly decreased by 43% (p<0,05) and by 28% (p<0,01), respectively. Meanwhile, the granular cells remained unaltered in experimental animals of both age groups. Decrease of the projection mitral neurons, in its turn, results in disruption of the projections to olfactory cortex and hampers proper functioning of the olfactory analyzer. PMID- 16705244 TI - Modulation of no synthesis by plaferon LB in liver. AB - Nitric oxide (NO) is a free radical involved in the regulation of many physiological processes. NO exerts important effects on liver in health and diseases. In the present study, we attempted to evaluate the influence of plaferon LB on the liver injury in two experimental models: immune-mediated and mechanical (traumatic shock) liver damage models. The results of our study demonstrated that plaferon LB decreased levels of NO in traumatic shock, but enhanced NO expression on con-A induced hepatitis. Furthermore, plaferon LB inhibited development of metabolic changes in liver tissue and facilitated restoration of hepatocyte function and structure of the tissue. Thus, our data revealed that plaferon LB was a powerful regulator of NO levels in experimental liver injury. Further investigations are needed to determine pro- and anti apoptotic properties of plaferon LB in the liver. PMID- 16705245 TI - [The principles of protection of work and workers' health of Tbilisi metro in modern social-economic conditions]. AB - At the present stage of development of the country, at the time of total social economical changes, providing the employed part of the population with safe work conditions and qualified health protection system plays an important role and it needs corresponding legislative basis. In the system of these measures the most important thing is organizing a comprehensive research with the aim of studying the work conditions and the state of health of workers engaged in national economy and working out complex sanitary actions. The comprehensive research has manifested the unfavourable state of factors of work and industrial environment (mental and emotional exertion, high level of dustiness). This exerts a significant influence on the functional state of Tbilisi metro workers. Cardiovascular and nervous system pathologies are the main factors in the state of health. On the grounds of clinico-hygienic comparison they can be related to industrial conditional diseases. On the basis of conducted the complex research recommendations about the improvement of work factors and the health state of Tbilisi metro workers (particularly those of organizational character) have been developed. PMID- 16705246 TI - [Changes of blood paramagnetic centres of animals irradiated with low-intensity laser]. AB - Therapeutic effects of lasers are based on activation of oxidative process on cellular and subcellular levels. The first photoacceptor of laser beam being mitochondria., which simultaneously represents the source of oxidation products as well as their target and thus, laser exposure can cause numerical effects: inactivation of electron chain components of mitochondria, energy metabolism inhibition, oxidation of lipids and DNA molecule. The aim of the given work is to investigate the influence of submaximal dozes of infrared (0,85 mkm) low intensity laser on the activity of oxidative processes in laboratory mice blood, which are reflected on the state of paramagnetic centers. For this purpose the condition of blood paramagnetic centers (caeruloplazmin, Fe(3+)-transferin, Fe(2+), Mn(2+), MetHb and NO) has been studied. Results imply that irradiation of mice blood with submaximal dozes of low-intensity laser causes the activation of oxidative process, but those changes do not lead to impairment of blood antioxidant features. PMID- 16705247 TI - Anti-aging peptide bioregulators induce reactivation of chromatin. AB - The effect of synthetic peptide bioregulators (Epitalon, Livagen and Vilon) on structural and facultative heterochromatin of cultivated lymphocytes have been studied among old (75-88yr.) people. The data obtained indicate that epitalon, livagen and vilon: 1) activate synthetic processes, caused by reactivation of ribosomal genes as a result of deheterochromatinization (decondensation) of nucleolus organizer regions; 2) induce unrolling (deheterochromatinization) of total heterochromatin; 3) release genes repressed by heterochromatinization (condensation) of euchromatic regions forming facultative heterochromatin; 4) epitalon and livagen induce deheterochromatinization (decondensation) of pericentromeric structural heterochromatin of the chromosomes1 and 9. However, vilon does not induce deheterochromatinization of pericentromeric structural heterochromatin. These results indicate that peptide bioregulators Epitalon, Livagen and Vilon cause activation (deheterochromatinization) of chromatin in lymphocytes of old individuals. PMID- 16705248 TI - Treatment of toxic liver damage by antihepatocytotoxic serum. AB - Acute liver failure (ALF) is a broad term that refers to both fulminant hepatic failure (FHF) and subfulminant hepatic failure (or late-onset hepatic failure). The latter term is reserved for patients with liver disease for up to 26 weeks prior to the development of hepatic encephalopathy. Some patients with previously unrecognized chronic liver disease decompensate and present with liver failure; although this technically is not FHF, discerning this at the time of presentation may not be possible (eg, Wilson disease). The objective of present research was to define suitable method for the treatment of toxic damage of the liver. We consider that liver changes occurred in the case of toxic damage can be reversed by administration of antihepatocytotoxic serum which has been developed by us. For this purpose we are going to induce liver toxic damage in the Wister line and investigate the liver regeneration cellular mechanisms after administration of antihepatocytotoxic serum. Experimental investigations were performed on Wister line male rats, animals were divided into four equal groups. In I and II groups we were modelling acute liver failure by injections of hepatotoxic agent CCL4 (I) and performing 70% hepatectomy (II), III group was served as control group, and IV group served as donors for HPCs. The main aspect of our study was to stimulate liver reparative regeneration and by this help to organ function restoration. Performed studies have shown hypertrophy and moderate hyperplasia of hepatocyte organelles. Oval shaped HPCs were also observed, Performed investigations had shown effectiveness of our ALF treatment method in terms of damaged liver function restoration, normalization of morphological picture and biochemical measurements and we hope that administration of antihepatocytotoxic serum developed by us can give a chance to patients with acute liver failure. PMID- 16705249 TI - [The modulatory influence of moxonidine on the different link of sympatho-adrenal system and hemodynamic reactions in normo and hypertensive rats during emotional stress]. AB - Effect of moxonidine on hemodynamic parameters, baroreflex sensitivity (BS) and sympatho-adrenal system was studied on the model of emotional stress in normotensive (N) and hypertensive (H) rats. Moxonidine was administered intravenously in daily dose 100 mcg/kg during one week prior to stress condition. It was shown that moxonidine reduced pressor reactions of arterial pressure, tachycardia and have increased BS especially in H animals, which was blunted during aversive stimulation. Moxonidine also revealed the effect of pharmacological correction of catecholamine - norepinephrine and epinephrine content in the myocardium and adrenals in stress and post stress period more markedly in H rats. The data obtained indicate the preventive antistress activity of the moxonidine. PMID- 16705250 TI - [Modern view on biochemistry of atherosclerosis]. AB - The article summarizes the literature data on clinical chemistry of pathogenesis of atherosclerosis. According to the up-to-date view, atherogenic are considered the products of chemical modification of LDL, rather than LDL themselves. The modification is created by means of the free radicals or reactive oxidised nitrogen species (RONS). Their rapid and uncontrollable generation in the body may become a prerequisite for the development of a number of diseases and pathological processes, such as atherosclerosis, neurological, malignancies, aging and inflammation and etc... The article describes the chemical nature of free radicals, the mechanism of their action and chain character of their generation. The particular attention is paid to nitric oxide, which is recognized in a broad array of biologic systems, namely, actions on vascular endothelium and mediating macrophage activity. The mechanisms of cell protection from the toxic action of RONS have been elucidated. Based on the experimental data presented, extremely large doses of antioxidants may lead to health problems, rather than confer benefits, because RONS are involved in the mechanisms which increase the survival of cells at unfavourable conditions. The complete impairment of their generation promotes the weakening of cell immunity. The article describes the means to provide cellular cholesterol homeostasis and the uptake of chemically modified LDL by macrophage scavenger receptors. Macrophages consume excess modified lipoprotein becoming foam cells. Foam cells accumulate, releasing growth factors and cytokines that stimulate the migration of smooth muscle cells from the media to the intima, where they proliferate, produce collagen and take up lipid, potentially becoming foam cells which are the main culprits of atherosclerotic changes in the artery walls. PMID- 16705251 TI - [Quantitative analysis of psychometric indicators among 40-55 years old accelerant Georgian men and women of normal physical development]. AB - Determination of individual characteristics of a human being causes much interest. Furthermore, you can seldom come across the studies about determination of psychological categories of accelerants. The aim of the present work was to determine quantitative analysis of psychometric indicators among 40-55 years old accelerant Georgian men and women of normal physical development for given age scales as well as to determine personal characteristics of each person to be researched. On the basis of conducted research we may shape out, that older Georgian women are characterized with phlegmatic temperament and introversive character. They are observed to have average degree of logic and verbal intellect; constitutional types of mid exciting, conflicting and static mood. While elder Georgian men have sanguine temperament, they are characterized with extroversive character, average degree of logic intellect, plastic-dynamic and constitutional-stable mood; they are considered to be harmonic and dynamic constitutional types. We have also established that Georgian accelerant men are observed to have phlegmatic temperament, extroversion, average mathematical intellect and conflicting and static mood. Accelerant Georgian women displayed phlegmatic temperament, extroversion, average logic intellect, and conflicting and static mood. But still they were observed, though seldom, to have harmonic and dynamic character. PMID- 16705252 TI - Comments on 'The influence of cardiac preload and positive end-expiratory pressure on the pre-ejection period'. AB - The pre-ejection period (PEP) has been described as a potential parameter for monitoring cardiac preload in deeply sedated mechanically ventilated patients. Other authors have recently suggested that PEP is not sensitive to the changes in intravascular volume status in mechanically ventilated pigs which underwent acute hemorrhage. The present comment is an analysis of this recent animal investigation. PMID- 16705254 TI - Sublingual capnometry: a non-invasive measure of microcirculatory dysfunction and tissue hypoxia. AB - With improvement in supportive care patients rarely die from their presenting illness but rather from its sequela, namely sequential multi-organ failure. Tissue hypoxia is believed to be the causation of multi-organ dysfunction syndrome (MODS). The expedient detection and correction of tissue hypoxia may therefore limit the development of MODS. The standard oxygenation and hemodynamic variables (blood pressure, arterial oxygenation, cardiac output) which are monitored in critically ill patients are 'upstream' markers and provide little information as to the adequacy of tissue oxygenation. Global 'downstream' markers such as mixed venous oxygen saturation and blood lactate are insensitive indicators of tissue hypoxia. Sublingual PCO(2) is a regional marker of microvascular perfusion and tissue hypoxia that holds great promise for the risk stratification and end-point of goal directed resuscitation in critically ill patients. This paper reviews the technology and application of sublingual PCO(2) monitoring. PMID- 16705255 TI - A dynamic method to estimate the time split between the A2 and P2 components of the S2 heart sound. AB - The time interval between the aortic (A2) and the pulmonary (P2) components of the second heart sound (S2) is an indicator of pulmonary arterial pressure. However, knowledge of the A2 and P2 components of the S2 sound is difficult to obtain due to their temporal overlap and significant spectral similarity. In this work, we aim to extract the A2 and P2 components from the phonocardiogram to estimate the time interval between them. We attain our objective by first isolating the S2 sound from the phonocardiogram by utilizing the mode complexity of the heart. Then, we assume the statistical independence of the A2 and P2 components and extract them from the S2 sound by the application of blind source separation techniques. Once separated, the time interval between the A2 and P2 components is estimated with a time-centroid-based method. Experimental results using simulated data show excellent performance of the proposed algorithm to extract the A2 and the P2 components from the S2 sound and to estimate the time interval between them. Results obtained from real data are also encouraging and show promise for utilizing the proposed method in a clinical setting to non invasively tract pulmonary hypertension. PMID- 16705257 TI - Electrical impedance spectroscopy and the diagnosis of bladder pathology. AB - Bladder pathology is usually investigated visually by cystoscopy. At present, definitive diagnosis of the bladder can be made by biopsy only, usually under general anaesthesia. This is a relatively high-cost procedure in terms of both time and money and is associated with discomfort for the patient and morbidity. Thus, we used an electrical impedance spectroscopy technique for differentiating pathological changes in the urothelium and improving cystoscopic detection. For ex vivo study, a whole or part of the patient's urinary bladder was used to take the readings less than half an hour after excision at room temperature, about 27 degrees C, using the Mk3.5 Sheffield System (2-384 kHz in 24 frequencies). In this study, 145 points (from 16 freshly excised bladders from patients) were studied in terms of their biopsy reports matching to the electrical impedance measurements. For in vivo study, a total of 106 points from 38 patients were studied to take electrical impedance and biopsy samples. The impedance data were evaluated in both malignant and benign groups, and revealed a significant difference between these two groups. The impedivity of the malignant bladder tissue was significantly higher than the impedivity of the benign tissue, especially at lower frequencies (p < 0.001). In addition, the receiver operating characteristic (ROC) curve for impedance measurements indicated that this technique could provide diagnostic information (individual classification is possible). Thus, the authors have investigated the application of bio-impedance measurements to the bladder tissue as a novel and minimally invasive technique to characterize human bladder urothelium. Therefore, this technique, especially at lower frequencies, can be a complementary method for cystoscopy, biopsy and histopathological evaluation of the bladder abnormalities. PMID- 16705256 TI - Centre-specific multichannel electrogastrographic testing utilizing wavelet-based decomposition. AB - Although the principles of electrogastrography (EGG) have been known for years, the clinical utility of EGG has not been clearly demonstrated, and EGG recording and analysis techniques have not been fully standardized. The aim of this study was to develop a multichannel EGG technique for detecting abnormal gastric motility using an EGG database specifically designed for a particular testing centre, maximizing the sensitivity and the specificity of the test. Eight healthy volunteers formed a reference group to which 4 study groups (17 gastro oesophageal reflux disease (GORD) patients, 7 functional dyspepsia patients, 8 post-fundoplication patients and 12 healthy volunteers) were compared. Eight channel EGG was recorded in the postprandial and fasting states for 30 min each. The recorded signals were wavelet compressed and the resulting error (per cent root mean square difference (PRD)) after the compression was utilized to compare the study groups to the reference group. A threshold in the number of channels with significantly different PRD values was introduced. Sensitivity (SE), specificity (SP) and correct classification rate (CC) of the test in recognizing each clinical condition in the study groups for several channel thresholds and compressions were calculated, and were maximized. Increasing the compression and channel threshold levels improved the specificity, but decreased the sensitivity of the multichannel EGG test. An optimal combination region was identified based on a centre-specific adjustment of the channel threshold and the wavelet compression. The achieved maximum sensitivity, specificity and correct classification for this region in our test centre were as follows: GORD--SE 82.4%, SP 83.3%, CC 82.8%; functional dyspepsia--SE 100%, SP 75%, CC 84.2%; post fundoplication--SE 75.0%, SP 83.3%, CC 80.0%. The utilization of a wavelet-based decomposition technique to process multichannel EGG signals can be a very effective method for enhancing the clinical utility of EGG, provided it is specifically developed for a given testing centre. PMID- 16705258 TI - Assessment of electrocardiogram visual interpretation strategy based on scanpath analysis. AB - Automated ECG interpretation systems are supposed to follow human expert reasoning. Despite well-established standards, the visual interpretation strategy of the human is still undisclosed today. This paper presents a new approach to the interpretation process research based on eyetrack features captured from a human expert during biosignal visual inspection. This approach required a set of visual tasks consisting in ECG interpretation by volunteers of different degrees of expertise. The recorded eyeglobe trajectories were analysed in the context of medical data represented in the displayed ECG traces and revealed interesting information on diagnostic data distribution and principles of interpretation strategies. The scanpath-derived data make benefit of oculomotoric habits gathering in everyday practice, unconscious mutual perception-recognition interactions and are not affected by human memory or verbalization limits. For these reasons, they provide more objective assessment than any other method willingly controlled by the human. Besides new information about the ECG contents and quantitative descriptions of medical data distribution, our experiment reveals some eyetrack parameters as distinctive for interpretation skills estimation. PMID- 16705259 TI - Reliability and sensitivity of jerk-cost measurement for evaluating irregularity of chewing jaw movements. AB - Rate of change in movement acceleration (jerk) is a universally accepted quantity to evaluate irregularities of human limb and eye movements. This study was aimed to attest reliability of the jerk measurement of masticatory jaw movements and to identify its sensitivity to discriminate between movements performed with the presence of varied occlusal interference. Jaw movements during gum chewing were recorded with a 3D tracking device. Twelve adult subjects participated in the experiments. For five subjects, normalized jerk-costs (NJC) during jaw closing were compared between those measured on two separate occasions. For seven subjects, the NJCs during closing were compared with those measured with/without introduction of four different types of occlusal interference. The NJCs did not differ significantly between the two recording occasions. The interference at the canine tooth induced greater increase in the NJCs than that at the molar tooth. The comparison between repeated measures revealed reproducibility of the NJCs. In addition, the NJC was shown to be capable of discriminating between irregularities of the movements induced by occlusal interference at the canine and molar tooth sites. These findings suggest that the NJC is a valid indicator of masticatory jaw movement irregularity induced by disturbances of full intercuspation between upper and lower teeth. PMID- 16705260 TI - Storage effects on the Cole-Cole parameters of erythrocyte suspensions. AB - Chemical alterations of red blood cells (RBCs) during storage eventually affect the electrical properties of blood. In this study, the physiological parameters such as extracellular (SAGM + CPD + residual plasma) Na(+), K(+), Cl(-), pH, 2,3 DPG and ATP together with the Cole-Cole parameters were measured using erythrocyte suspensions from 51 male donors (31 donors form the training set and 20 donors are used for testing), on the 0th, 10th, 21st, 35th and 42nd days of storage. During storage, while the surrounding fluid resistance (R(e)) and the effective cell membrane capacitance (C(m)) increased progressively with time, the intracellular fluid resistance (R(i)) has decreased. Storage of RBCs resulted in a rise in K(+) and a fall in Na(+), Cl(-), pH, 2,3-DPG and ATP. Accordingly, electrical parameters were all correlated with Na(+), K(+), Cl(-), pH and ATP at varying levels. By applying multi-regression analysis, it is concluded that R(i), R(e) and C(m) are appropriate for modeling Na(+), K(+), Cl(-), pH and ATP during storage. PMID- 16705261 TI - A longitudinal study of methanol in the exhaled breath of 30 healthy volunteers using selected ion flow tube mass spectrometry, SIFT-MS. AB - Selected ion flow tube mass spectrometry, SIFT-MS, has been used to monitor the volatile compounds in the exhaled breath of 30 volunteers (19 male, 11 female) over a six-month period. Volunteers provided breath samples each week between 8:45 am and 1 pm (before lunch), and the concentrations of several trace compounds were obtained. In this paper the focus is on methanol in breath. The median methanol level determined using the H(3)O(+) precursor ions for all samples was 461 parts-per-billion (ppb), the concentrations for all the samples ranging from 32 to 1684 ppb. The distribution of breath methanol concentration is seen to be log-normal for this healthy population; the geometric mean was 450 ppb, close to the median value, and the multiplicative (geometric) standard deviation was 1.62. Breath methanol is not correlated with age, breath ethanol or ethanol consumed in the previous 24 h, but there was an inverse correlation with body mass index (BMI) for the cohort of volunteers recruited for this study. Observed breath methanol levels are well compatible with the previously published blood methanol levels. Some tentative suggestions are made concerning the origin of endogenous methanol. PMID- 16705262 TI - The effect of local mild cold exposure on pulse transit time. AB - Pulse transit time (PTT) is a simple, noninvasive technique which shows great promise in the continuous monitoring of blood pressure and the assessment of arterial stiffness, and has potential applications in wearable healthcare devices. Usually, PTT is measured from the electrocardiogram (ECG) R-wave to a characteristic point on the peripheral pulse by photoplethysmography. However, peripheral blood circulation is sensitive to local temperature variation. This study investigated the effect of mild cold exposure on PTT by immersing one finger into cold water. The photoplethysmographic (PPG) signals were recorded from the cooled and adjacent uncooled fingers. The significant change in dc and ac amplitudes of the PPG pulse indicates that the mild cold exposure has a substantial effect on finger blood circulation. Three measurements of PTT were performed by calculating the time delay from the ECG R-wave to three different characteristic points on the PPG pulse, namely, the foot (PTT1), the point on the rising limb with maximal slope (PTT2) and the peak (PTT3), respectively. PTT3 was found to be easily subject to waveform distortion. Significant changes in the difference of PTT1 and PTT2 between two fingers were observed after a 5 min recovery period. The changes in PTT1 and PTT2 on the reference finger also showed close correlation (r = -0.77 and r = -0.80, p < 0.001) with the changes in SBP after recovery, whereas those measured on the test finger have low correlation (r = -0.53 and r = -0.38, p > 0.15). The results suggest that mild cold exposure may have a delay effect on PTT due to cold-induced vasodilatation and could be a potential source of error. The effect of cold exposure should be carefully examined, especially when PTT that includes a large portion of rising time of peripheral pulse is applied. PMID- 16705264 TI - An improved asynchronous brain interface: making use of the temporal history of the LF-ASD feature vectors. AB - The low-frequency asynchronous switch design (LF-ASD) has been introduced as a direct brain interface (BI) for asynchronous control applications. Asynchronous interfaces, as opposed to synchronous interfaces, have the advantage of being operational at all times and not only at specific system-defined periods. This paper modifies the LF-ASD design by incorporating into the system more knowledge about the attempted movements. Specifically, the history of feature values extracted from the EEG signal is used to detect a right index finger movement attempt. Using data collected from individuals with high-level spinal cord injuries and able-bodied subjects, it is shown that the error characteristics of the modified design are significantly better than the previous LF-ASD design. The true positive rate percentage increased by up to 15 which corresponds to 50% improvement when the system is operating with false positive rates in the 1-2% range. PMID- 16705263 TI - Photometric recording of transmembrane potential in outer hair cells. AB - Cochlear outer hair cells (OHCs) are polarized epithelial cells that have mechanoelectrical transduction channels within their apical stereocilia and produce electromotile force along their lateral wall. Phase shifts, or time delays, in the transmembrane voltage occurring at different axial locations along the cell may contribute to our understanding of how these cells operate at auditory frequencies. We developed a method to optically measure the phase of the OHC transmembrane potential using the voltage-sensitive dye (VSD) di-8-ANEPPS. The exit aperture of a fibre-optic light source was driven in two dimensions so that a 24 microm spot of excitation light could be positioned along the length of the OHC. We used the whole-cell patch-clamp technique in the current-clamp mode to stimulate the OHC at the base. The photometric response and the voltage response were monitored with a photodetector and patch-clamp amplifier, respectively. The photometric response was used to measure the regional changes in the membrane potential in response to maintained (dc) and sinusoidal (ac) current stimuli applied at the base of the cell. We used a neutral density filter to lower the excitation light intensity and reduce phototoxicity. A sensitive detector and lock-in amplifier were used to measure the small ac VSD signal. This permitted measurements of the ac photometric response below the noise floor of the static fluorescence. The amplitude and phase components of the photometric response were recorded for stimuli up to 800 Hz. VSD data at 400-800 Hz show the presence of a small phase delay between the stimulus voltage at the base of the cell and the local membrane potential measured along the lateral wall. Results are consistent with the hypothesis that OHCs exhibit inhomogeneous membrane potentials that vary with position in analogy with the voltage in nerve axons. PMID- 16705266 TI - Chronic histological effects of the flat interface nerve electrode. AB - The flat interface nerve electrode (FINE) is designed to reshape peripheral nerves into favorable geometries for selective stimulation. Compared to cylindrical geometries, the ovoid geometries created by the FINE allow more space for contact placement. Furthermore, the amount of electrically excitable tissue between stimulating contacts and target axons is reduced. In this study, the nerve response to the presence of the FINE is examined histologically. Three different FINEs were designed to reshape peripheral nerves to different opening heights designated as 'wide' (1.3 mm), 'medium' (0.5 mm) and 'narrow' (0.1 mm) cuffs. Twelve adult cats were implanted with one cuff each (four in total of each design) on their right sciatic nerves. At least 3 months later, the animals were sacrificed and their sciatic nerves were harvested for histological evaluation. Cross-sectional areas and eccentricities (defined as the major axis divided by the minor axis of the closest fit ellipse to a region) of the nerves were measured to assess the degree of reshaping. The wide and medium cuff designs significantly reshaped the nerves compared to control nerves, though there was no significant difference in eccentricity between nerves implanted with wide and medium cuffs. There was extensive deposition of connective tissue in the epineurium of all nerves implanted with cuffs. No significant difference in fiber counts was measured in any of the groups studied. Only nerves implanted with narrow cuffs showed evidence of axonal injury and/or demyelination. These results, coupled with stimulation selectivity measurements made on the same animals, suggest that safe, selective electrodes can be designed with ovoid geometries. Moderate reshaping caused no damage, while extreme reshaping generated mild-to-moderate nerve damage. It might be possible, however, to redesign the cuffs to slowly reshape the nerves. PMID- 16705265 TI - Compact self-wiring in cultured neural networks. AB - We present a novel approach for patterning cultured neural networks in which a particular geometry is achieved via anchoring of cell clusters (tens of cells/each) at specific positions. In addition, compact connections among pairs of clusters occur spontaneously through a single non-adherent straight bundle composed of axons and dendrites. The anchors that stabilize the cell clusters are either poly-D-lysine, a strong adhesive substrate, or carbon nanotubes. Square, triangular and circular structures of connectivity were successfully realized. Monitoring the dynamics of the forming networks in real time revealed that the self-assembly process is mainly driven by the ability of the neuronal cell clusters to move away from each other while continuously stretching a neurite bundle in between. Using the presented technique, we achieved networks with wiring regions which are made exclusively of neuronal processes unbound to the surface. The resulted network patterns are very stable and can be maintained for as long as 11 weeks. The approach can be used to build advanced neuro-chips for bio-sensing applications (e.g. drug and toxin detection) where the structure, stability and reproducibility of the networks are of great relevance. PMID- 16705267 TI - Design, in vitro and in vivo assessment of a multi-channel sieve electrode with integrated multiplexer. AB - This paper reports on the design, in vitro and in vivo investigation of a flexible, lightweight, polyimide based implantable sieve electrode with a hybrid assembly of multiplexers and polymer encapsulation. The integration of multiplexers enables us to connect a large number of electrodes on the sieve using few input connections. The implant assembly of the sieve electrode with the electronic circuitry was verified by impedance measurement. The 27 platinum electrodes of the sieve were coated with platinum black to reduce the electrode impedance. The impedance magnitude of the electrode sites on the sieve (geometric surface area 2,200 microm(2)) was |Z(f=1kHz)| = 5.7 kOmega. The sieve electrodes, encased in silicone, have been implanted in the transected sciatic nerve of rats. Initial experiments showed that axons regenerated through the holes of the sieve and reinnervated distal target organs. Nerve signals were recorded in preliminary tests after 3-7 months post-implantation. PMID- 16705268 TI - Magnetoneurographic registration of propagating magnetic fields in the lumbar spine after stimulation of the posterior tibial nerve. AB - Stimulation of the posterior tibial nerve has been associated with different somatosensory evoked potentials (SEP) recorded along the spine and thorax. The aim of this study was to register and describe the magnetic fields corresponding to different components of spinal SEP after stimulation of tibial nerves. In nine healthy subjects, right and left posterior tibial nerves were transcutaneously electrostimulated at the ankles. Neuromagnetic fields were registered over a circular 800 cm(2) area of the lumbosacral spine using a 61-channel biomagnetometer. Magnetic field maps were constructed and examined visually for dipolar patterns. Equivalent current dipoles (ECD) were calculated for each somatosensory evoked field (SEF) using a least-squares fit in a spherical model. In seven subjects dipolar SEF were detected over the lower back at two separate latencies and locations and propagating ECD could be localized. Both the first and second components found agreed anatomically and functionally with respect to propagation in the underlying nerve fibers. It was possible to record and identify SEF which correspond to the SEP described in the literature. Dipole localization based on an equivalent current dipole model allowed a basic evaluation of the plausibility of the measurements with respect to the processes being examined. PMID- 16705269 TI - The effect of cystic cavities on deep brain stimulation in the basal ganglia: a simulation-based study. AB - Although the therapeutic effect of deep brain stimulation (DBS) is well recognized, a fundamental understanding of the mechanisms responsible is still not known. In this study finite element method (FEM) modelling and simulation was used in order to study relative changes of the electrical field extension surrounding a monopolar DBS electrode positioned in grey matter. Due to the frequently appearing cystic cavities in the DBS-target globus pallidus internus, a nucleus of grey matter with and without a cerebrospinal fluid filled cystic cavity was modelled. The position, size and shape of the cyst were altered in relation to the electrode. The simulations demonstrated an electrical field around the active element with decreasing values in the radial direction. A stepwise change was present at the edge between grey and white matters. The cyst increased the radial extension and changed the shape of the electrical field substantially. The position, size and shape of the cyst were the main influencing factors. We suggest that cystic cavities in the DBS-target may result in closely related unexpected structures or neural fibre bundles being stimulated and could be one of the reasons for suboptimal clinical effects or stimulation-induced side effects. PMID- 16705270 TI - Different classification techniques considering brain computer interface applications. AB - In this work the application of different machine learning techniques for classification of mental tasks from electroencephalograph (EEG) signals is investigated. The main application for this research is the improvement of brain computer interface (BCI) systems. For this purpose, Bayesian graphical network, neural network, Bayesian quadratic, Fisher linear and hidden Markov model classifiers are applied to two known EEG datasets in the BCI field. The Bayesian network classifier is used for the first time in this work for classification of EEG signals. The Bayesian network appeared to have a significant accuracy and more consistent classification compared to the other four methods. In addition to classical correct classification accuracy criteria, the mutual information is also used to compare the classification results with other BCI groups. PMID- 16705271 TI - A comparison of optimal MIMO linear and nonlinear models for brain-machine interfaces. AB - The field of brain-machine interfaces requires the estimation of a mapping from spike trains collected in motor cortex areas to the hand kinematics of the behaving animal. This paper presents a systematic investigation of several linear (Wiener filter, LMS adaptive filters, gamma filter, subspace Wiener filters) and nonlinear models (time-delay neural network and local linear switching models) applied to datasets from two experiments in monkeys performing motor tasks (reaching for food and target hitting). Ensembles of 100-200 cortical neurons were simultaneously recorded in these experiments, and even larger neuronal samples are anticipated in the future. Due to the large size of the models (thousands of parameters), the major issue studied was the generalization performance. Every parameter of the models (not only the weights) was selected optimally using signal processing and machine learning techniques. The models were also compared statistically with respect to the Wiener filter as the baseline. Each of the optimization procedures produced improvements over that baseline for either one of the two datasets or both. PMID- 16705272 TI - Selection and parameterization of cortical neurons for neuroprosthetic control. AB - When designing neuroprosthetic interfaces for motor function, it is crucial to have a system that can extract reliable information from available neural signals and produce an output suitable for real life applications. Systems designed to date have relied on establishing a relationship between neural discharge patterns in motor cortical areas and limb movement, an approach not suitable for patients who require such implants but who are unable to provide proper motor behavior to initially tune the system. We describe here a method that allows rapid tuning of a population vector-based system for neural control without arm movements. We trained highly motivated primates to observe a 3D center-out task as the computer played it very slowly. Based on only 10-12 s of neuronal activity observed in M1 and PMd, we generated an initial mapping between neural activity and device motion that the animal could successfully use for neuroprosthetic control. Subsequent tunings of the parameters led to improvements in control, but the initial selection of neurons and estimated preferred direction for those cells remained stable throughout the remainder of the day. Using this system, we have observed that the contribution of individual neurons to the overall control of the system is very heterogeneous. We thus derived a novel measure of unit quality and an indexing scheme that allowed us to rate each neuron's contribution to the overall control. In offline tests, we found that fewer than half of the units made positive contributions to the performance. We tested this experimentally by having the animals control the neuroprosthetic system using only the 20 best neurons. We found that performance in this case was better than when the entire set of available neurons was used. Based on these results, we believe that, with careful task design, it is feasible to parameterize control systems without any overt behaviors and that subsequent control system design will be enhanced with cautious unit selection. These improvements can lead to systems demanding lower bandwidth and computational power, and will pave the way for more feasible clinical systems. PMID- 16705273 TI - Optimizing neurotrophic factor combinations for neurite outgrowth. AB - Most neurotrophic factors are members of one of three families: the neurotrophins, the glial cell-line derived neurotrophic factor family ligands (GFLs) and the neuropoietic cytokines. Each family activates distinct but overlapping cellular pathways. Several studies have shown additive or synergistic interactions between neurotrophic factors from different families, though generally only a single combination has been studied. Because of possible interactions between the neurotrophic factors, the optimum concentration of a factor in a mixture may differ from the optimum when applied individually. Additionally, the effect of combinations of neurotrophic factors from each of the three families on neurite extension is unclear. This study examines the effects of several combinations of the neurotrophin nerve growth factor (NGF), the GFL glial cell-line derived neurotrophic factor (GDNF) and the neuropoietic cytokine ciliary neurotrophic factor (CNTF) on neurite outgrowth from young rat dorsal root ganglion (DRG) explants. The combination of 50 ng ml(-1) NGF and 10 ng ml( 1) of each GDNF and CNTF induced the highest level of neurite outgrowth at a 752 +/- 53% increase over untreated DRGs and increased the longest neurite length to 2031 +/- 97 microm compared to 916 +/- 64 microm for untreated DRGs. The optimum concentrations of the three factors applied in combination corresponded to the optimum concentration of each factor when applied individually. These results indicate that the efficacy of future therapies for nerve repair would be enhanced by the controlled release of a combination of neurotrophins, GFLs and neuropoietic cytokines at higher concentrations than used in previous conduit designs. PMID- 16705274 TI - High frequency electrical conduction block of the pudendal nerve. AB - A reversible electrical block of the pudendal nerves may provide a valuable method for restoration of urinary voiding in individuals with bladder-sphincter dyssynergia. This study quantified the stimulus parameters and effectiveness of high frequency (HFAC) sinusoidal waveforms on the pudendal nerves to produce block of the external urethral sphincter (EUS). A proximal electrode on the pudendal nerve after its exit from the sciatic notch was used to apply low frequency stimuli to evoke EUS contractions. HFAC at frequencies from 1 to 30 kHz with amplitudes from 1 to 10 V were applied through a conforming tripolar nerve cuff electrode implanted distally. Sphincter responses were recorded with a catheter mounted micro-transducer. A fast onset and reversible motor block was obtained over this range of frequencies. The HFAC block showed three phases: a high onset response, often a period of repetitive firing and usually a steady state of complete or partial block. A complete EUS block was obtained in all animals. The block thresholds showed a linear relationship with frequency. HFAC pudendal nerve stimulation effectively produced a quickly reversible block of evoked urethral sphincter contractions. The HFAC pudendal block could be a valuable tool in the rehabilitation of bladder-sphincter dyssynergia. PMID- 16705275 TI - Notes from the board: President's message. PMID- 16705277 TI - In search of advanced clinical nurse specialist education. PMID- 16705279 TI - "Smart" infusion pumps and the clinical nurse specialist. PMID- 16705280 TI - Nosocomial infections and bath water: any cause for concern? PMID- 16705281 TI - Strategies for success: bringing evidence-based practice to the bedside. PMID- 16705282 TI - Managing the pain of postherpetic neuralgia. PMID- 16705283 TI - Entrepreneurship: is it in your DNA? PMID- 16705284 TI - The PhD-prepared nurse in the clinical setting. AB - BACKGROUND: The emergence of new doctoral programs within the nursing discipline has stimulated dialogue regarding the role of the doctorally prepared nurse in the clinical setting. National nursing organizations have cited the need for additional research that would provide information regarding the current practice of doctorally prepared nurses. A review of the literature reveals little published information about the role of PhD nurses in the clinical setting. PURPOSE: The purpose of this descriptive qualitative study was to investigate how PhD-prepared nurses describe and define their role within the clinical setting. METHODS: Interviews were conducted with 5 PhD nurses who were all employed full time in the clinical setting. Interview notes were recorded, transcribed, and analyzed using qualitative data analysis. Recurrent themes that emerged from the interviews were identified. RESULTS: The 2 themes that emerged from the interviews were bridging the research/practice gap and serving as a healthcare leader. All participants spoke to their role in leading, encouraging, or participating in clinical nursing research within their healthcare institutions. Phrases regarding leadership emerged throughout each interview and reflected a number of leadership responsibilities that each participant had within the healthcare environment. CONCLUSIONS: The findings from this qualitative study provide insight into the current role of the PhD-prepared nurse in the clinical setting. This information can be used to guide additional research that might influence the development of future doctoral programs in nursing. PMID- 16705285 TI - Quality of life outcomes related to anemia management of patients with chronic renal failure. AB - PURPOSE: The purpose of this study was to evaluate the impact of a clinical nurse specialist-managed outpatient anemia management program on quality of life for patients with anemia related to chronic kidney disease. DESCRIPTION OF THE STUDY: A retrospective study was conducted using information from the Medical Outcomes Short Form 36 Item Health Survey, which is completed by patients with anemia at their initial, 3-month, 6-month, and 12-month visits, and annually thereafter. Thirty-four patients completed the 3-month survey, 19 completed the 6-month survey, and 10 completed the 12-month survey. OUTCOMES: There was a statistically significant increase in quality of life indicators at the 3 and 6 months' interval. The increase in physical and decrease in mental indicators were not substantiated through the 12-month interval. CONCLUSION: Quality of life was significantly improved for patients in a clinical nurse specialist-managed outpatient anemia management program. PMID- 16705287 TI - A CNS-facilitated ICD support group: a clinical project evaluation. AB - PURPOSE: The purpose of this article is to describe and evaluate a clinical nurse specialist (CNS)-facilitated support group for recipients of implantable cardioverter defibrillator. Specific evaluation aims were as follows: (1) How do demographic and clinical factors differ between those who attended the support group and those who did not? (2) Is there a difference in the quality of life index (QLI) of individuals with an implantable cardioverter defibrillator who attended the CNS-facilitated support group and those who did not? (3) What demographic and clinical factors are related to QLI? DESIGN AND METHOD: Clinical project theory-based objectives were described. Implementation of the project was evaluated by retrospective survey of all implantable cardioverter defibrillator recipients during a 10-year time frame using the Ferrans and Powers' Quality of Life Index: Cardiac Version and demographic questionnaire. Attendance sheets defined who attended so comparisons could be made. EVALUATION RESULTS: One hundred and twelve surveys were returned (34% return rate). A positive relationship between CNS visit during hospitalization, number of and value of supports, years of education, and ejection fraction was noted with attendance at the support group. No between-group differences on total QLI, or on any subscales, were found. Comorbidity was the only clinical factor correlated with QLI. CONCLUSION: CNS-facilitated support groups can be offered as an additional support. Evaluation design issues limited the measuring outcomes of existing interventions. Future prospective studies are recommended to determine the affect of the support group on quality of life. PMID- 16705289 TI - Nursing research: thoughts on professional obligation, discipline and knowledge management. PMID- 16705290 TI - Promoting excellence in nursing research and scholarship in the clinical setting. PMID- 16705304 TI - Maximizing the utility of interorganizational data using concept mapping. AB - OBJECTIVE: The overall objective of this article was to apply an existing methodology (concept mapping) to a nonstandardized interorganizational data set. The specific aims were to (1) identify, define, and create a map that represented the common conceptual domains of patient satisfaction; (2)validate the relationships among concepts; and (3) use the map by testing the relationships of the patient satisfaction concepts to other patient outcomes. BACKGROUND: The lack of standardized methodologies for collecting patient outcome data across multiple institutions poses threats to the validity and generalizability of research findings. METHODS: The steps in concept mapping were used to explicate the common underlying conceptual dimensions from 3 patient satisfaction tools. The map was then used to evaluate the extent that patient satisfaction was related to outcomes of hospitalized patients. Each of 3 hospitals' measure of patient satisfaction varied in the number and type of items. All items were examined to identify potential areas of conceptual correspondence. RESULTS: Items were grouped into 1 of the 3 identified categories that were consistent across sites: caring, communication, and responsiveness. Moderate correlations were found among the concepts of satisfaction and medication errors, nosocomial infections, and patient falls. CONCLUSIONS: Concept mapping-more traditionally used for learning, project planning, and evaluation-is a technique that has demonstrated utility in multi-institutional research. PMID- 16705305 TI - The evolution of an ambulatory nursing intensity system: measuring nursing workload in a day hospital setting. AB - OBJECTIVE: To establish a methodology for developing a user-friendly, environmentally specific nursing intensity measurement tool. BACKGROUND: Nursing intensity measurement systems assess the demand for nursing care in a given setting and must adapt to changes in technology, patient populations, and treatment profiles. Nowhere is this more evident than in a clinical research setting where demands for nursing care are driven by changing patient populations and new biomedical research protocols. This article traces the extension of an adult oncology ambulatory nursing intensity system at the Clinical Center, National Institutes of Health, into 3 day hospitals: medical surgical, pediatrics, and hematology/oncology. METHODS: A team of stakeholders including nurse managers, clinical nurse specialists, and senior clinical research nurses was assembled. Descriptions of clinical research care for each day hospital were developed and organized into 6 levels with their associated time frames. Descriptions were reviewed by 2 content specialists, and a content validity index quantified the extent of their agreement. A weighted kappa statistic was used to measure interrater reliability. RESULTS: A content validity index of .87 indicated a high level of agreement between specialists. The weighted kappa statistic was .80, demonstrating substantial agreement between 2 raters. CONCLUSION: This ambulatory nursing intensity tool proved to be adaptable to heterogeneous patient populations. PMID- 16705306 TI - The positive face of resistance: nurses relate their stories. AB - OBJECTIVE: To relate nurses' stories of their experiences of acts of resistance. BACKGROUND: Although resistance is often characterized as negative, it can be seen as a way that the less powerful speak up about workplace concerns. By studying how nurses resist, new perspectives about how some groups exercise power may be understood. METHODS: For this qualitative study, 19 nurses were interviewed. They were asked to describe an act of resistance in their professional life. The interviews were tape recorded, transcribed, and then analyzed using narrative methods. RESULTS: Four major categories emerged from the narratives: definitions of resistance, relationships, core narratives, and creating meaning. This article focuses on 2 of these for space constraints and relevance to this journal: relationships and creating meaning. CONCLUSION: The nurses interviewed described situations in their work lives, where they felt that they performed an act of resistance because of unfair treatment, abuse of power, or ethical concerns. It often took courage to do so, but the acts had mainly positive effects on them and their institutions. It is theorized that resistance may actually be positive for both the staff nurse and the organization. PMID- 16705308 TI - Predictors of sustained organizational commitment among nurses with temporary job contracts. AB - OBJECTIVE: To examine sociodemographic, work-related factors and psychological health as predictors of sustained organizational commitment among temporary hospital employees. BACKGROUND: The participants were 412 nurses who had a temporary job contract and reported being committed to their organization at baseline. METHODS: Organizational commitment was measured again 2 years later. RESULTS: The results of logistic regression analysis showed that age over 35 years, high job control, high participative safety, high perceived justice in decision making, and low psychological distress predicted sustained organizational commitment at follow-up. The change from temporary employment to a permanent job and high job control predicted sustained organizational commitment even after the effect of all the other predictors was taken into account. CONCLUSION: Organizations that employ temporary workers should pay attention to the job control and career prospects of temporary staff. PMID- 16705307 TI - The impact of nursing work environments on patient safety outcomes: the mediating role of burnout/engagement. AB - OBJECTIVE: To test a theoretical model of professional nurse work environments linking conditions for professional nursing practice to burnout and, subsequently, patient safety outcomes. BACKGROUND: The 2004 Institute of Medicine report raised serious concerns about the impact of hospital restructuring on nursing work environments and patient safety outcomes. Few studies have used a theoretical framework to study the nature of the relationships between nursing work environments and patient safety outcomes. METHODS: Hospital-based nurses in Canada (N = 8,597) completed measures of worklife (Practice Environment Scale of the Nursing Work Index), burnout (Maslach Burnout Inventory-Human Service Scale), and their report of frequency of adverse patient events. RESULTS: Structural equation modeling analysis supported an extension of Leiter and Laschinger's Nursing Worklife Model. Nursing leadership played a fundamental role in the quality of worklife regarding policy involvement, staffing levels, support for a nursing model of care (vs medical), and nurse/physician relationships. Staffing adequacy directly affected emotional exhaustion, and use of a nursing model of care had a direct effect on nurses' personal accomplishment. Both directly affected patient safety outcomes. CONCLUSIONS: The results suggest that patient safety outcomes are related to the quality of the nursing practice work environment and nursing leadership's role in changing the work environment to decrease nurse burnout. PMID- 16705309 TI - Differences in perceptions of empowerment among nationally certified and noncertified nurses. AB - OBJECTIVE: To examine the difference in perceptions of empowerment between nurses who were nationally certified and those who were not. BACKGROUND: Nurses are encouraged to obtain certification in their specialty. The focus of most nursing research on certification has been on motivation to acquire certification and perceived benefits of certification. Research related to empowerment indicates that access to empowerment structures results in achievement and success. METHODS: This descriptive comparative study used the Conditions of Work Effectiveness II Questionnaire to measure registered nurses' perceptions of empowerment. Certification status was recorded on the demographic section of the questionnaire. RESULTS: Significant differences were noted in empowerment scores for certified and noncertified nurses as measured by the Conditions of Work Effectiveness II Questionnaire. Findings suggest that nurses who are certified have higher perceptions of empowerment. Certification may increase nurses' perceptions of empowerment and therefore improve work effectiveness. CONCLUSION: Certified nurses in this study had increased access to job-related power and opportunity structures. Certification provides recognition of the nurses' knowledge and expertise in the specialty area which in turn is empowering. Organizations that support and recognize this achievement may experience improved turnover and retention rates. PMID- 16705310 TI - Phase 1 trial of the antiangiogenic peptide ATN-161 (Ac-PHSCN-NH(2)), a beta integrin antagonist, in patients with solid tumours. AB - To evaluate the toxicity, pharmacological and biological properties of ATN-161, a five -amino-acid peptide derived from the synergy region of fibronectin, adult patients with advanced solid tumours were enrolled in eight sequential dose cohorts (0.1-16 mg kg(-1)), receiving ATN-161 administered as a 10-min infusion thrice weekly. Pharmacokinetic sampling of blood and urine over 7 h was performed on Day 1. Twenty-six patients received from 1 to 14 4-week cycles of treatment. The total number of cycles administered to all patients was 86, without dose limiting toxicities. At dose levels above 0.5 mg kg(-1), mean total clearance and volume of distribution showed dose-independent pharmacokinetics (PKs). At 8.0 and 16.0 mg kg(-1), clearance of ATN-161 was reduced, suggesting saturable PKs. Dose escalation was halted at 16 mg kg(-1) when drug exposure (area under the curve) exceeded that associated with efficacy in animal models. There were no objective responses. Six patients received more than four cycles of treatment (>112 days). Three patients received 10 or more cycles (> or =280 days). ATN-161 was well tolerated at all dose levels. Approximately, 1/3 of the patients in the study manifested prolonged stable disease. These findings suggest that ATN-161 should be investigated further as an antiangiogenic and antimetastatic cancer agent alone or with chemotherapy. PMID- 16705312 TI - Controlled clinical trials in cancer pain. How controlled should they be? A qualitative systematic review. AB - This qualitative systematic review of the clinical methodology used in randomised, controlled trials of oral opioids (morphine, hydromorphone, oxycodone) for cancer pain underlines the difficulties of good pain research in palliative care. The current literature lacks placebo-controlled superiority trials. Recommendations for future research are discussed. PMID- 16705311 TI - Differential disruption of cell cycle pathways in small cell and non-small cell lung cancer. AB - Lung cancer is the leading cause of cancer-related mortality in the world, with small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) comprising the two major cell types. Although these cell types can be distinguished readily at the histological level, knowledge of their underlying molecular differences is very limited. In this study, we compared 14 SCLC cell lines against 27 NSCLC cell lines using an integrated array comparative genomic hybridisation and gene expression profiling approach to identify subtype-specific disruptions. Using stringent criteria, we have identified 159 of the genes that are responsible for the different biology of these cell types. Sorting of these genes by their biological functions revealed the differential disruption of key components involved in cell cycle pathways. Our novel comparative combined genome and transcriptome analysis not only identified differentially altered genes, but also revealed that certain shared pathways are preferentially disrupted at different steps in these cell types. Small cell lung cancer exhibited increased expression of MRP5, activation of Wnt pathway inhibitors, and upregulation of p38 MAPK activating genes, while NSCLC showed downregulation of CDKN2A, and upregulation of MAPK9 and EGFR. This information suggests that cell cycle upregulation in SCLC and NSCLC occurs through drastically different mechanisms, highlighting the need for differential molecular target selection in the treatment of these cancers. PMID- 16705313 TI - Securin (hPTTG1) expression is regulated by beta-catenin/TCF in human colorectal carcinoma. AB - Overexpression of the transcriptional activator beta-catenin, mostly owing to loss-of-function mutations of the adenomatous polyposis coli (APC) tumour suppressor gene, is crucial for the initiation and progression of human colorectal carcinogenesis. Securin is a regulator of chromosome separation and its overexpression has been shown to be involved in different tumour-promoting processes, like transformation, hyperproliferation and angiogenesis, and correlates with tumour cell invasion. However, the molecular mechanism leading to securin overexpression in human colorectal cancer is unknown. Here we show a correlated high expression of beta-catenin and securin (hPTTG1) in colorectal adenomas and carcinomas and further demonstrate that securin is a target of beta catenin transcriptional activation. This implies that deregulation of the beta catenin/T-cell factor-signalling pathway leads to overexpression of securin in human colorectal cancer, which subsequently may contribute to tumour progression. PMID- 16705314 TI - Potentiation of the anticancer effect of valproic acid, an antiepileptic agent with histone deacetylase inhibitory activity, by the kinase inhibitor Staurosporine or its clinically relevant analogue UCN-01. AB - Histone deacetylase inhibitors (HDACIs) are novel anticancer agents with potent cytotoxicity against a wide range of malignancies. We have previously demonstrated that either Calphostin C (CC) (a protein kinase C (PKC) inhibitor) or Parthenolide (an NF-kappaB inhibitor) abrogates HDACI-induced transcriptional activation of NF-kappaB and p21, which is associated with profound potentiation of HDACI-mediated induction of apoptosis. Valproic acid (VA), a commonly used antiepileptic agent, has recently been shown to be an HDACI. This study was aimed to evaluate the anticancer property of VA in thoracic cancer cells and the development of clinically relevant strategies to enhance VA-mediated induction of apoptosis using kinase inhibitors Staurosporine (STP) or its analogue UCN-01. Treating cultured thoracic cancer cells with VA (0.62-10.0 mM) resulted in significant cell line- and dose-dependent growth inhibition (IC(50) values: 4.1 6.0 mM) and cell cycle arrest at G1/S checkpoint with profound accumulation of cells at G0/G1 phase but little induction of apoptosis. Valproic acid, being an HDACI, caused significant dose-dependent accumulation of hyperacetylated histones, following 24 h of treatment. Valproic acid-mediated 5-20-fold upregulation of transcriptional activity of NF-kappaB was substantially (50-90%) suppressed by cotreatment with CC, STP or UCN-01. Whereas minimal death (<20%) was observed in cells treated with either VA (1.0 or 5.0 mM) alone or kinase inhibitors alone, 60-90% of cells underwent apoptosis following exposure to combinations of VA+kinase inhibitors. Kinase inhibitor-mediated suppression of NF kappaB transcriptional activity played an important role in sensitising cancer cells to VA as direct inhibition of NF-kappaB by Parthenolide drastically synergised with VA to induce apoptosis (VA+Parthenolide: 60-90% compared to <20% following single-drug treatments). In conclusion, VA, a well-known antiepileptic drug, has mild growth-inhibitory activity on cultured cancer cells. The weak VA mediated induction of apoptosis of thoracic cancer cells can be profoundly enhanced either by Parthenolide, a pharmacologic inhibitor of NF-kappaB, or by UCN-01 a kinase inhibitor that has already undergone phase I clinical development. Combinations of VA with either a PKC inhibitor or an NF-kappaB inhibitor are promising novel molecularly targeted therapeutics for thoracic cancers. PMID- 16705316 TI - Radiochemotherapy in stage III/IV nonmetastatic squamous cell head and neck cancer. PMID- 16705315 TI - A prospective study of Kaposi's sarcoma-associated herpesvirus and Epstein-Barr virus in adults with human immunodeficiency virus-1. AB - Antibody titres against Kaposi's sarcoma associated herpesvirus (KSHV or human herpesvirus 8 (HHV-8)) and Epstein-Barr virus (EBV) were examined in people who subsequently developed Kaposi's sarcoma and non-Hodgkin's lymphoma, within randomised controlled trials of antiretroviral therapy in adults infected with the human immunodeficiency virus-1 (HIV). For each case of Kaposi's sarcoma (n=189) and each case of non-Hodgkin's lymphoma (n=67), which developed after randomisation, one control was randomly selected from other trial participants, after matching for age, sex, ethnicity, mode of HIV transmission, type of treatment received and period of follow-up. Using sera taken an average of two and a half years before the diagnosis of cancer, titres of antibodies against KSHV latent (LANA) and lytic (K8.1) antigens and against EBV (VCA) antigens were investigated in relation to subsequent risks of cancer by calculating odds ratios (OR) using conditional logistic regression. Latent antibodies against KSHV were detectable among 38% (72 out of 189) of Kaposi's sarcoma cases and 12% (23 out of 189) of their controls (OR=4.4, 95% confidence intervals (CI) 2.3-8.3, P<0.001). The OR for Kaposi's sarcoma increased with increasing antilatent KSHV antibody titre (chi(2)(1) for trend=32.2, P<0.001). Lytic antibodies against KSHV were detectable among 33% (61 out of 187) of Kaposi's sarcoma cases and 19% (36 out of 187) of their controls (OR=2.0, 95% CI 1.2-3.4, P=0.003) and the OR for Kaposi's sarcoma increased with increasing antilytic KSHV antibody titre (chi(2)(1) for trend=6.2, P=0.02). Virtually, all cases and controls had anti-EBV antibodies detected and the OR for non-Hodgkin's lymphoma associated with a doubling of the anti-EBV antibody titre was estimated to increase by a multiplicative factor of 1.3 (95% CI 0.9-1.7, P=0.1). Kaposi's sarcoma was not associated with antibody levels against EBV (P=0.4) and non-Hodgkin's lymphoma was not associated with antibodies against KSHV (latent P=0.3; lytic P=0.5). Adjustment for CD4 count at the time of sample collection made no material difference to any of the results. In conclusion, among human immunodeficiency virus infected people, high levels of antibodies against KSHV latent and lytic antigens are strongly associated with subsequent risk of Kaposi's sarcoma but not non-Hodgkin's lymphoma. Antibody titre to EBV does not appear to be strongly associated with subsequent risk of Kaposi's sarcoma or non-Hodgkin's lymphoma in HIV infected people. PMID- 16705320 TI - Sex determination and inbreeding depression in an ant with regular sib-mating. AB - Haplodiploidy is one of the most widespread mechanisms of sex determination in animals. In many Hymenoptera, including all hitherto investigated social species, diploid individuals, which are heterozygous at the sex locus, develop as females, whereas haploid, hemizygous individuals develop as males (single-locus complementary sex determination, sl-CSD). Inbreeding leads to homozygosity at the sex locus, resulting in the production of diploid males, which are usually sterile and constitute a considerable fitness cost. Nevertheless, regular inbreeding without diploid male production is known from several solitary wasps, suggesting that in these species sex is not determined by sl-CSD but alternative mechanisms. Here, we examine sex determination in an ant with regular inbreeding, Cardiocondyla obscurior. The almost complete absence of diploid males after 10 generations of brother-sister mating in the laboratory documents for the first time the absence of sl-CSD and CSD with two or a few unlinked sex loci in a species of social Hymenoptera. Queens, which mated with a brother, appeared to decrease the number of males in their brood, as expected from the relatedness relationships under inbreeding. In contrast, some colonies began to show signs of an inbreeding depression after several generations of sib-mating, such as shortened queen life span, higher brood mortality, and a shift to more male biased sex ratios in some colonies, presumably due to lower insemination capability of sperm. PMID- 16705321 TI - Frequency and fitness cost of resistance to Bacillus thuringiensis in Chrysomela tremulae (Coleoptera: Chrysomelidae). AB - The "high dose-refuge" (HDR) strategy is commonly recommended and currently used for delaying or preventing pest adaptation to transgenic plants producing Bacillus thuringiensis (Bt) toxins. The efficiency of this strategy depends, among other factors, on the initial frequency of Bt resistance alleles and on the fitness costs associated with these alleles. Two years ago, an allele conferring resistance to Bt poplar was detected in a French population of the poplar pest Chrysomela tremulae F. Although this pest had never been subjected to Bt selection pressure due to human activities, the frequency of this allele was estimated at 0.0037, with a 95% credible (CI) interval of 0.00045-0.0080. We investigated the frequency of this allele in a second sample of C. tremulae collected more than 500 km from the site of the initial population. The estimated frequency in this sample was 0.0113 (95% CI 0.0031-0.0247), reinforcing the conclusion that resistance to Bt plants may be present at detectable frequencies in pest populations before selection resulting from pest management by humans. The frequency of the Bt resistance allele over the two samples was 0.0049 (95% CI 0.0020-0.0091). We also followed five laboratory lines in which the frequency of this allele was initially fixed at 0.500. After five generations maintained on non-Bt poplar leaves, the frequency of this allele decreased in all lines, whereas allelic frequencies at a neutral locus were unaffected. Thus, the Bt resistance allele detected in French populations of C. tremulae is probably associated with a fitness cost. PMID- 16705322 TI - Speciation: Splitting when together. PMID- 16705323 TI - Genetic consequences of human management in an introduced island population of red deer (Cervus elaphus). AB - We investigated phylogeography and spatial genetic structure in an introduced island population of red deer (Cervus elaphus) on the Isle of Rum, Scotland, experiencing spatial variation in management regime. Five different mitochondrial DNA (mtDNA) haplotypes were present among female red deer on Rum. These comprised two phylogenetically divergent groups, one of which clustered with red deer from Sardinia and North Africa, while the other four grouped with other Western European red deer. Recent and historical red deer management practices explain this result. The Rum population is descended from recent introductions from at least four different UK mainland populations, and translocation of red deer within the UK and across Europe is well documented. We found significant spatial genetic structure across Rum in both mtDNA haplotypes and microsatellite markers. Mitochondrial spatial structure was over an order of magnitude greater than structure in nuclear markers. This extreme difference is explained by the fact that the Rum population was introduced from different source populations, the highly male-biased dispersal patterns of red deer and the much smaller effective population size of mitochondrial compared to nuclear markers. Spatial structure in mtDNA conformed to a pattern of isolation by distance, while nuclear DNA did not. Apparent structure in the nuclear markers was driven by differences between the North Block and the rest of the island. We suggest that recent differences in the management regimes in different parts of the island have led to differences in effective male migration that would account for this observation. PMID- 16705324 TI - Genetic structure of the forest pest Hylobius abietis on conifer plantations at different spatial scales in Europe. AB - The distribution of genetic variation within and among 20 European sites infested by the large pine weevil, Hylobius abietis, was analysed using dominant amplified fragment length polymorphism markers. Analysis of molecular variance was performed at the European, regional and local scales. Most of the genetic variability was found within rather than among populations and the global fixation index averaged over loci was low (0.07). We found no evidence of genetic drift, even in relatively isolated sites. This genetic pattern tends to confirm the high dispersal ability of the weevil and the influence of human-mediated expansion of its range through conifer plantations across Europe since the 19th century. Assignment tests demonstrated that the regional forest is a pertinent geographic scale for defining populations in the large pine weevil. Testing the potential influence of the larval host-plant identity (Scot Pine vs Norway Spruce) on the genetic structure revealed a weak but significant effect in two of the three regions tested (in Ardeche and in Limousin but not in Finland). One locus varied with host-plant use in the two French regions, indicating a potential role in host-plant adaptation. However, host-race formation is not observed in H. abietis; we discuss this result in the light of our current knowledge of this insect's biology. Altogether, this study shows that the use of different host plants for development does not constitute a strong barrier to gene flow for H. abietis and confirms the high dispersal ability of this forest pest. PMID- 16705325 TI - [Populations aging and hospital productivity]. PMID- 16705326 TI - [The academic surgeon - a species in risk of extinction?]. PMID- 16705327 TI - [Results of Dor's endoventricular repair in the management of left ventricular aneurysms]. AB - INTRODUCTION: Left ventricular aneurysms are serious long-term complications from acute myocardial infarction that produce hemodynamic alterations in the cardiac function. OBJECTIVE: The aim of this study is to evaluate the chirurgical and clinic results of the endoventricular patch repair of the akinetic or dyskinetic portions of the left ventricle. METHODS: From August 2002 to October 2005, 20 consecutive patients underwent surgical repair of left ventricular aneurysm using the endoventricular patch repair technique. The mean age was 63,6 years, and 80% where male. Every patient have had only one episode of acute myocardial infarction, which resulted in a akinetic or dyskinetic alteration in the motility of the left ventricle. The preoperative functional class of most of the patients was class III or IV and 45% where on angina class II. In 90% of the patients the ventricular aneurysm where antero-septal. Left ventricular dysfunction was present in 65%. RESULTS: All patients underwent the Dor procedure associated with coronary artery bypass grafting. There was no perioperative mortality. One patient died in the immediate postoperative period. Four patients needed inotropic support for more than 24h and intra-aortic balloon pumping was used postoperatively in two cases. Mean hospital stay was 8,6 days. At late follow-up the functional class was I in 20%, class II in 55% and class III in 10%. The angina class was I in 65% of the cases. CONCLUSIONS: The surgical repair of left ventricular aneurysm using the endoventricular patch repair technique proved to be safe, causing significant clinical improvement and an increase in the ejection fraction. PMID- 16705328 TI - [Transaortic resection of left ventricular fibroelastoma]. AB - Left ventricular tumors are extremely rare. Complete surgical resection is usually curative and recurrence is rare. The left atrial approach is safe and advisable when possible. We report a rare case of left ventricular tumor in a 71 year-old man presented with a cerebellar stroke. A two-dimensional echocardiogram revealed a mobile mass with the stalk attached deeply in left ventricular wall. To improve surgical access, a transversal aortotomy was performed and the mass was gently pulled and excised. Intracavitary left ventricular tumors, which are deeply situated, difficult to see, and inaccessible via left atriotomy, may be made surgically accessible by a transaortic approach. A vetriculotomy should be avoided. PMID- 16705329 TI - [Cervical lymphadenopathy as first manifestation of a bronchogenic cyst]. AB - A 26-year old woman was admitted with clinical and laboratory signs and symptoms of chronic infection and multiple cervical lymphadenopathy, whose pathological studies disclosed a non-specific inflammatory reaction. Anti-tuberculosis treatment was then started, with no significant improvement. Chest CT scans was compatible with a mediastinial tumor and RMN studies revealed the presence of an infected bronchogenic cyst, which was surgically and successfully removed with complete disappearance of the infection. The authors conclude by enhancing the fact that bronchogenic cysts may course with diverse clinical manifestations and should be included in differential diagnosis of mediastinal tumors. PMID- 16705330 TI - [The role of protocols as indicator of "process" in a Department of Anesthesiology-example of an anesthetic protocol for lung transplantation]. AB - This paper is aimed at to demonstrate and enhance the importance of anesthetic protocols in general sense, and for the lung transplantation in particular, as markers of "process" in the perioperative anesthetic care. The authors describe and discuss the main steps of the organization of a specific protocol dedicated to lung transplantation and put particular emphasis on the need of a good integration of the different specialties and professional groups directly involved in this demanding field of activity. PMID- 16705331 TI - [Justa-renal aortic aneurysms repair: consequences of the supra-renal aortic clamping, based on an experience of 44 operated cases]. AB - The present study is aimed at the evaluation of the consequences of the supra renal aortic clamping in the surgical management of justa-renal aortic aneurysms, based on a personal experience of 44 cases operated on for the last 25 years. The series includes 42 men (95.5%) and 2 women (4.5%) with an average age of 70 years. All patients were operated on transperitonealy and the transection and reconstruction of left renal vein was made in 15 instances. Supra-renal aortic clamping and renal average ischemic time was 13'3 minutes, and no significant functional renal alterations could be noticed, nor the need for hemodialysis support. Contrarily, significant occlusive lesions of the renal arteries (4 instances) or the superior mesenteric artery (1 case) resulted as a mechanical consequences of the supra-renal aortic clamping. Three cases were recognized and repaired intraoperatively, through renal revascularization, one case was not diagnosed, coursed with anuric renal failure and death (mortality 2.2%). Patient with mesenteric artery occlusion developed an acute mesenteric ischemia which was successfully managed conservatively. The author concludes by enhancing the risks of the mechanical consequences arising from the supra-renal aortic clamping, namely renal or mesenteric ischemia. Its prompt recognition and repair seems mandatory, in order to avoid the severe and even fatal consequences that it may cause, raising the morbi-mortality rates that sometimes accompanies the surgical management of justa-renal aortic aneurysms. PMID- 16705332 TI - [Supra-genicular femoro-popliteal bypass: 5 years retrospectively analysis]. AB - We report a retrospective study of patients submitted to supragenicular femoro popliteal bypass surgery in our department between 1998 and 2002. A SPSS package was used for statistical analysis. Eighty bypasses were performed in 74 patients with a median follow up of 19,6 months [1-71 (+/- 22,5 months)]. Leriche-Fontaine stage IV chronic ischemia was the main indication for surgery accounting for 68,8% of cases, followed by stage III (25%) and stage IIb (6,2%). The most prevalent vascular conduit was PTFE (87,5%), with great saphenous vein and Dacron being used on 7,5 % and 5% of cases. Ten patients (12,5 %) later required major amputation and limb salvage at 12, 24 and 36 months was 91,6 %, 87,7 % and 82,8 %. The primary patency rates were 81,7 %, 78,9 % and 71 % at the end of the first, second and third year of follow up. Patient survival was 92,8 %, 92,8 % e 88,6 % at 12, 24 e 36 months of follow up. A large proportion of patients (61,7 %) required an accessory procedure. Our results might be considered acceptable in face of other published results, though care must be taken given the intrinsic limitations of this retrospective study. PMID- 16705334 TI - [Thrombotic occlusion of the terminal aorta associated to a renal fusion and pelvic location ("pancake" kidney). First case report]. AB - Coexistence of thrombotic occlusion of the terminal aorta with a "pancake" kidney is reported in 63-year old man, diagnosed by CT-scans and selective angiography, to assess the renal circulation and for surgical planning. Surgical exposure was transperitoneal and revascularization was accomplished through a bifurcation Dacron graft from the supraceliac aorta to both femorals, associated to the renal revascularization, by means of PTFE graft from the prosthesis to the renal artery. The success of procedure was later demonstrated by angio-RM, showing both grafts working in excellent condition. According to an extensive bibliographic review, this seems to be the first case reporting the association of a thrombotic occlusion of the terminal aorta to a "pancake" kidney, thus justifying its presentation and divulgation. PMID- 16705333 TI - [Revascularization to the midpopliteal artery through a posterior approach. Two case reports]. AB - An original technique consisting in the revascularization of the midpopliteal artery through a posterior approach is described, in patients with critical ischemia of the lower extremities, utilizing a prosthetic substitute. This technique can be regarded as an alternative to the infragenicular revascularization and it is particularly suited for patients with no autologous material available. Two successful cases operated on with this technique are reported, with a 6 months and 1 year follow up and documented patency of the grafts. PMID- 16705335 TI - [Leiomyosarcoma of the inferior vena cava. Clinical report]. AB - A leiomyosarcoma of the inferior vena cava is reported in a 52 year old female, located in the para-renal segment of the vena cava. The tumor was resected and the vena cava was replaced by a 20 mm PTFE anelar prosthesis, with reimplantation of the right renal vein. The left renal vein was ligated. One year later CT-scans showed patency of the graft. The authors discuss the therapeutic options for this type of tumor and analyse the 5 and 10 years survival rates, following treatment. PMID- 16705336 TI - [Bronchial artery aneurysm]. PMID- 16705338 TI - Porous coordination polymers with zeolite topologies constructed from 4-connected building units. AB - Two novel 3D coordination polymers, Cd(CTC)(H2O).(H2PIP)(0.5)(H2O) (1) with zeolite ABW topology and Cd(CTC).(HIPA) (2) with zeolite BCT topology, have been synthesized by constructing inorganic and organic 4-connected building units and using the organic bases as templates, and the frameworks of and not only expand the original structures of zeolites ABW and BCT, but also exhibit significant advantages over them in terms of thermal stability, ion exchange and adsorption. PMID- 16705337 TI - Pi-bonded quinonoid transition-metal complexes. AB - Coordination of the carbocyclic ring of hydroquinones to electrophilic transition metal fragments such as Mn(CO)3+ and Rh(COD)+ produces stable pi-bonded eta6 complexes that are activated to facile reversible deprotonation of the -OH groups. The deprotonations are accompanied by electron transfer to the transition metal, which acts as an internal oxidizing agent or electron sink. With manganese as the metal, the resulting eta5-semiquinone and eta4-quinone complexes have been used to synthesize one- two- and three-dimensional polymeric metal-organometallic coordination networks. With rhodium as the metal, the pi-quinonoid complexes have been demonstrated to play a unique role in multifunctional C-C coupling catalysis and in the synthesis of new organolithium reagents. Both classes of pi-quinonoid complexes appear to have significant applications in nanochemistry by providing an excellent vehicle for templating the directed self-assembly of nanoparticles into functional materials. PMID- 16705339 TI - Density functional theory investigation of the geometric and electronic structures of [UO2(H2O)m(OH)n](2 - n) (n + m = 5). AB - Gradient corrected density functional theory has been used to calculate the geometric and electronic structures of the family of molecules [UO2(H2O)m(OH)n](2 - n) (n + m = 5). Comparisons are made with previous experimental and theoretical structural and spectroscopic data. r(U-O(yl)) is found to lengthen as water molecules are replaced by hydroxides in the equatorial plane, and the nu(sym) and nu(asym) uranyl vibrational wavenumbers decrease correspondingly. GGA functionals (BP86, PW91 and PBE) are generally found to perform better for the cationic complexes than for the anions. The inclusion of solvent effects using continuum models leads to spurious low frequency imaginary vibrational modes and overall poorer agreement with experimental data for nu(sym) and nu(asym). Analysis of the molecular orbital structure is performed in order to trace the origin of the lengthening and weakening of the U-O(yl) bond as waters are replaced by hydroxides. No evidence is found to support previous suggestions of a competition for U 6d atomic orbitals in U-O(yl) and U-O(hydroxide)pi bonding. Rather, the lengthening and weakening of U-O(yl) is attributed to reduced ionic bonding generated in part by the sigma-donating ability of the hydroxide ligands. PMID- 16705340 TI - The synthesis of new weakly coordinating diborate anions: anion stability as a function of linker structure and steric bulk. AB - The successive addition of KCN and Ph3CCl to B(C6F4-C6F5-2)3 (PBB) affords triphenylmethyl salts of the [NC-PBB]- anion. By contrast, the analogous reaction with sodium dicyanamide followed by treatment with Ph(3)CCl leads to the zwitterionic aminoborane H2NB(C12F9)2C12F8, via nucleophilic attack on an o-F atom, together with CPh3[F-PBB]. Whereas treatment of [NC-PBB]- with either PBB or B(C6F5)3 fails to give isolable cyano-bridged diborates, the reaction of Me3SiNC-B(C6F5)3 with PBB in the presence of Ph3CCl affords [Ph3C][PBB-NC B(C6F5)3]. Due to steric hindrance this anion is prone to borane dissociation. The longer linking group N(CN)2- gives the very voluminous anions [N[CNB(C6F5)3]2]- and [N(CN-PBB)2]-. A comparison of propylene polymerisations with rac-Me2Si(Ind)2ZrMe2 activated with the various boranes or trityl borates gives an anion-dependent activity sequence, in the order [NC-PBB]- < [MeB(C6F5)3] < [MePBB]- approximately [PBB-NCB(C6F5)3]- approximately [N[CNB(C6F5)3]2]- < [F PBB]-<< [B(C6F5)4]- < [N(CN-PBB)2]-. The anion [N(CN-PBB)2]- gives a catalyst productivity about 2500 times higher than that of [NC-PBB]- and exceeds that of [B(C6F5)4]- based catalysts. The van der Waals volumes and surface areas of the anions have been calculated and provide a rationale for the observed reactivity trends in polymerisation reactions. PMID- 16705341 TI - Mono- and binuclear complexes of iron(II) and iron(III) with an N4O ligand: synthesis, structures and catalytic properties in alkane oxidation. AB - Three mononuclear iron complexes and one binuclear iron complex, [Fe(tpoen)Cl].0.5(Fe2OCl6) (1), [Fe(tpoen)Cl]PF6 (2), Fe(tpoen)Cl3 (3) and [[Fe(tpoen)]2(mu-O)](ClO4)4 (4) (tpoen = N-(2-pyridylmethoxyethyl)-N,N-bis(2 pyridylmethyl)amine), were synthesized as functional models of non-heme iron oxygenases. Crystallographic studies revealed that the Fe(II) center of 1 is in a pseudooctahedral environment with a pentadentate N4O ligand and a chloride ion trans to the oxygen atom. The Fe(III) center of 3 is ligated by three nitrogen atoms of tpoen and three chloride ions in a facial configuration. Each Fe(III) center of 4 is coordinated with four nitrogen atoms and an oxygen atom of tpoen with the Fe-O-Fe angle of 172.0(3) angstroms. Complexes 2, 3 and 4 catalysed the oxidation of cyclohexane with H2O2 in the total TNs of 24-36 with A/K ratios of 1.9-2.4. Under the same conditions they also catalysed both the oxidation of ethylbenzene to benzylic alcohol and acetobenzene with good activity (30-47 TN) and low selectivity (A/K 0.7), and the oxidation of adamantane with moderate activity (15-18 TN) and low regioselectivity (3 degrees/2 degrees 3.0-3.2). With mCPBA as oxidant the catalytic activities of 2, 3 and 4 increased 1.8 to 2.3-fold for the oxidation of cyclohexane and ethylbenzene and 6.3 to 7.5-fold for the oxidation of adamantane. Drastic enhancement of the regioselectivity was observed in the oxidation of adamantane (3 degrees/2 degrees 18.5-30.3). PMID- 16705342 TI - Syntheses of supramolecular CuCN complexes by decomposing CuSCN: a general route to CuCN coordination polymers? AB - The solvothermal reaction of CuSCN with 1,2-bis(diphenylphosphino)ethane (dppe) yielded a coordination polymer, which was characterized to be a complex of CuCN and 1,2-bis(diphenylthiophosphinyl)ethane (dppeS2): [(CuCN)2(dppeS2)]n (1). The identification of complex 1 reveals that CuSCN was decomposed and the sulfur was transferred to dppe, and represents a new example of the transformation of inorganic sulfur to organic sulfur. The weak coordination interactions between CuCN and dppeS2 indicate that dppeS2 may be substituted by ligands with strong coordination ability. The ligand 2,4,6-tris(2-pyridyl)-1,3,5-triazine (tpt) was chosen as a substitute ligand. Three novel CuCN coordination polymers of tpt were synthesized and characterized: [Cu2(CN)2(tpt)]n (2) with a 3-D (10,3)-a network, [Cu2(CN)2(tpt)]n (3) and [Cu2(SCN)(CN)(tpt)]n (4) both with a 2-D (6,3) network, and only complex 2 can be obtained from CuCN directly. Interestingly, compounds 2 and 3 are genuine high-dimensional supramolecular isomers. During the syntheses of 2-4, single crystals of dppeS2 were isolated, which indicates it was substituted by tpt ligand and also confirmed the transformation of sulfur from CuSCN to dppe. The transformation of sulfur can be observed only when the temperature is relative high (>160 degrees C). At 140 degrees C, complex 5 containing only CuSCN was attained and no dppeS2 has been monitored in the resulting filtrate. PMID- 16705343 TI - 1,8-Dimethylnaphthalene-bridged diphosphine ligands: synthesis and structural comparison of their palladium complexes. AB - The synthesis of a new series of ligands with a 1,8-dimethylnaphthalene backbone is reported, 1,8-(R2PCH2)2C10H6, where R = (t)Bu 1 (dbpn), (i)Pr 2 (dippn), Cy 3 (dchpn) and Ph 4 (dphpn). The ligand 1 is structurally characterised by X-ray crystallography. A comparative structural study of the respective (diphosphine)Pd(dba) and (diphosphine)PdCl2 complexes is carried out, comparing the X-ray crystal structures of complexes 6, 7, 8, 10, 11 and 12. It is shown that the geometry at the metal is affected by not only ligand demands, but also by the palladium oxidation state and the electronic properties of the ligands. Two qualitative stability series are also identified: 9 < 10 < 11 approximately 12 is observed, and P2Pd(dba) complexes are more stable than the corresponding P2PdCl2 complexes towards opening of the chelate ring. It is also concluded that the bite angle is heavily influenced by the electron donating properties of the ligand. PMID- 16705344 TI - Triruthenium carbonyl clusters derived from chiral aminooxazolines: synthesis and catalytic activity. AB - Treatment of [Ru3(CO)12] with the chiral aminooxazolines (+)-2-amino-(4R)-phenyl 2-oxazoline (H2amphox), (+)-2-amino-(4R,5S)-indanyl-2-oxazoline (H2aminox) and (+)-2-(2-anilinyl)-(4R,5S)-indanyl-2-oxazoline (H2aninox) in THF at reflux temperature, affords the complexes [Ru3(mu-H)(mu3-kappa2-Hox-N,N)(CO)9] (H2ox = H2amphox, 1; H2aminox, 2) and [Ru3(mu-H)(mu-kappa2-Haninox-N,N)(CO)9] (3). In all cases, the activation of an N-H bond has occurred and the resulting amido fragment spans an edge of the metal triangle, while the N atom of the oxazoline ring is attached to the remaining metal atom (as in 1 and 2), or to one of the metal atoms of the bridged edge (as in 3). The use of 1-3 as catalyst precursors in the asymmetric hydrogen-transfer reduction of acetophenone and in the asymmetric cycloaddition of cyclopentadiene and acroleine is reported. PMID- 16705345 TI - Synthesis, crystal structure and properties of the semiconducting salts (TTF)2[Ni(dtcr)2] and (ET)2[Ni(dtcr)2] based on [Ni(dtcr)2] dianions (dtcr = dithiocroconate). AB - The first examples of CT salts based on [Ni(dtcr)2] dianions (1) (dtcr = dithiocroconate = 4,5-disulfanylcyclopent-4-ene-1,2,3-trionate), (TTF)2[Ni(dtcr)2] (TTF = tetrathiafulvalene) (2) and (ET)2[Ni(dtcr)2] [ET = bis(ethylenedithio)tetrathiafulvalene] (3) are reported. The redox-active dianion 1, containing oxo-groups in the periphery of the molecule, has been selected to investigate the role of the oxo-groups in promoting intermolecular interactions and hopefully their conducting properties. The salts 2 and 3 have been prepared by electrocrystallisation methods and 3 shows a semi-metallic behaviour: sigma = 1 x 10(-3) omega(-1) cm(-1) at room temperature, with a low activation energy 60 meV, while crystals of 2 were unsuitable for conductivity measurements. The X-ray structural characterisation shows an alternate dianion-(cation)2 stacking and the capability of the oxo-groups to promote interstack contacts. In 2, the TTF donors are present as face-to-face dimers of monocations (D2)2+. The stacking arrangement is different in 3, where ET monocations stack along two directions ([110] and [110]) in the same manner, with the repeating sequence (ET)-(ET) [Ni(dtcr)2]-(ET)-(ET) and are almost parallel to each other, with interplanar distances of 3.575 angstroms. Both structures are built on a dianion and two donor molecules, each one with a charge of +1. Diffuse reflectance combined with vibrational spectra complement structural results well. Crystal data: both 2 and 3 crystallise in the monoclinic space group P2(1)/c with a = 8.6340(8) angstroms, b = 21.586(2) angstroms, c = 7.5960(8) angstroms, beta = 95.625(11) degrees and V = 1408.9(2) angstroms3 for 2 and with a = 9.3700(7), b = 7.4410(6), c = 28.278(2) angstroms, beta = 99.039(6) degrees, V = 1947.1(3) angstroms3 for 3. PMID- 16705346 TI - Synthesis of layered (2-D) V-based bimetallic oxalates from non-aqueous media that cannot be synthesized from aqueous media. AB - The reaction of (NBu4)3[V(III)(ox)3] (1, ox = oxalate) and M(II) (M = Fe, Co, Ni, Cu) ions in MeCN, leads to the isolation of V-based coordination polymers of [N(n Bu)4][Fe(II)V(II)I(ox)3].0.30[[N(n-Bu)4](BF4)] (2), [N(n Bu)4][Co(II)V(III)(ox)3].0.75[[N(n-Bu)4](BF4)] (3), [N(n Bu)4][Ni(II)V(III)(ox)3].0.20[[N(n-Bu)4](BF4)].0.20MeCN (4), and [N(n Bu)4][Cu(II)V(III)(ox)2](BF4)2 (5) composition. Due to the lability of [V(III)(ox)3]3- to dissociate ox2-, these compounds cannot be prepared from aqueous media. 5 is best described as [N(n-Bu)4][V(III)Cu(II)(ox)2](BF4)2, and 2, 3, 4, and 5 are proposed to have a layered (2-D) motif for the MM(ox)x (x = 2, 3) extended framework. The [V(III)Cu(II)(ox)2] composition of 5 is reported for the first time for a bimetallic oxalate. 2 shows a weak antiferromagnetic interaction between Fe(II), S = 2 and V(III), S = 1 ions (theta = -9.4 K) within the 2-D layers. 3 and 5 do not magnetically order above 2 K. 4 magnetically order as ferromagnets below 2.55 K [taken as the onset of magnetization in chi''(T)], and has a glass transition temperature (chi'(max) at 1000 Hz) at 2.26 K. PMID- 16705347 TI - Synthesis, structural characterization and the first electroluminescent properties of tris- and bis-cycloiridiated complexes of sterically hindered electron-poor 2-(3,5-bis(trifluoromethyl)phenyl)-4-trifluoromethylpyridine. AB - An application of the new sterically hindered electron-poor 2-(3,5 bis(trifluoromethyl)phenyl)-4-trifluoromethylpyridine [HC--N] (1) in the one-step high temperature cyclometalation by Ir(III)Cl3 in the presence of Ag(I)OC(O)CF3 resulted in the synthesis of tris-cyclometalated complexes [C--N]2Ir[C--C] (3) and [C--N]3Ir (5). A neutral silver cluster with a repeating unit of hexa-silver groups in an infinite chain of (2) was isolated from the above reaction as well. When this cyclometalation was carried out in trimethylphosphate at lower temperature, bis-cyclometalated derivatives [C--N]2Ir(mu-Cl)2Ir[CN]2 (6), [C- N]2Ir[eta2-(O(C((t)Bu))2CH] (7), and [C--N]2Ir(mu-O-P(OMe)2-O)2Ir[C--N]2 (8) were synthesized. According to X-ray analyses complex (3), while trivalent, contains four cyclometalated single Ir-C bonds. One of the Ir-C bonds, next to the nitrogen atom of the CC pyridinium ligand, was found to be the shortest to date (1.977(4) angstroms) for a single bond between iridium and carbon atoms. The coordination of the C--C ligand in (3) to iridium has a decidedly interesting bonding pattern and can be explained by various formulations. The first one is considering this ligand as a monoanionic chelating ligand, in which the second coordination site arises from a carbene or azomethine ylide. Overall the best single picture may be a dianionic ligand making two normal Ir-C bonds, in which the ligand just happens to contain a pyridinium function that compensates for one negative charge on the iridium. LEDs constructed with compounds (7) and (8) give blue-green emission with peak electroluminescent efficiency of 15 and 2 cd A(-1), respectively. An LED constructed with compound (5) gives a yellowish emission with peak electroluminescent efficiency of 5.5 cd A(-1). PMID- 16705348 TI - The competition between acetate and pyrazolate in the formation of polynuclear Zn(II) coordination complexes. AB - Hydrated zinc(II) acetate reacts with pyrazole (Hpz) and, depending on the reaction conditions, forms different pyrazole-containing species, i.e. [[Zn(CH3COO)(mu-pz)(Hpz)]2] (1), [[Zn(CH3COO)2(Hpz)2.CH3COOH]] (2), [[Zn(mu pz)2]n] (3), and [[Zn(mu-CH3COO)(mu-pz)]n] (4). Their structural models have been derived from single-crystal X-ray diffractometry as well as from less conventional ab-initio X-ray powder methods. All species contain tetrahedrally coordinated Zn(II) ions, with Zn-N and Zn-O bond distances close to 2.0 angstroms. The existence of the [Zn(mu-pz)]2 core in the species 1, 3 and 4 indicates the propensity for the formation, in the presence of pyrazolate ligands, of well-defined dinuclear entities (with Zn...Zn contacts in the range 3.6-3.8 angstroms). The latter can mutually interact, in the crystals, through either hydrogen-bonding of ancillary ligands (as in 1) or coordinative bonds (via acetates, as in 4, or by self-complementarity, as in 3). The interconversion paths among these species have been studied, employing chemical and thermal methods. In particular, the topotactic and quantitative transformation of 1 into 3 by moderate heating is likely based on a solid-state cooperative condensation mechanism of the dangling pyrazolates toward neighbouring zinc(II) ions, with concomitant acetic acid extrusion. PMID- 16705349 TI - [Recurrent corneal erosion. Different treatment options with the excimer laser with emphasis on aggressive PTK]. AB - Recurrent corneal erosions in both traumatic and basal membrane dystrophy associated conditions seem to show a similar pathogenesis for abnormal adhesion between corneal epithelium and stroma. The purpose of each conservative or surgical treatment is fast healing without any infections or recurrences. This paper compares the different excimer laser treatment options with emphasis on the so-called aggressive PTK. A total of 50 eyes of 50 patients with recurrent corneal erosions after mechanical trauma or with epithelial basement membrane dystrophy were treated with aggressive PTK after the second or third recurrence. Aggressive PTK was performed with an ablation of 10-15 microm. After the mean follow-up time of 41.8 months, a recurrence rate of 6% without a change of visual acuity and a hyperopic shift of +0.74 D were observed. The earliest recurrent erosion showed up after 33 and the latest after 65 months. In comparison to other forms of PTK such as transepithelial or conventional subepithelial PTK, aggressive PTK showed the lowest recurrence rate reported for a long-term follow up. PMID- 16705350 TI - [Responsibilities of the otorhinolaryngologist in the follow-up of malignancies of the head and neck]. AB - The aims of follow-up of head and neck cancer patients are the detection of new tumour manifestations, management of impairments after tumour therapy, psychological care and the evaluation of therapeutic efficacy. The extent, success and cost-benefit ratio of follow-up are currently under discussion. We recommend interdisciplinary cooperation between the relevant specialists, such as oncology and radiotherapy, together with the otorhinolaryngologist for reasons of cost-efficacy and improvement of long-term results. We present a follow-up schedule for patients with head and neck squamous cell carcinomas, which are the by far most common manifestation. We recommend a standardized protocol, which should be individualized depending on tumour site, size, treatment and therapeutic options in the case of tumour recurrence. The most common salivary gland malignancies are also discussed. The objective is to increase the efficacy of follow-up in patients with head and neck cancer. PMID- 16705354 TI - A cytological study on Kelloggia (Rubiaceae), an intercontinental disjunct genus between eastern Asia and western North America. AB - A cytological study was carried out for the first time on four populations of the only two species of Kelloggia (Rubiaceae), which occur disjunctly in eastern Asia (K. chinensis Franch.) and western North America (K. galioides Torr.). The consistent mitotic prophase chromosome condensation pattern and interphase nuclei type were determined for both species. The chromosome base number of the genus is suggested to be x = 11. The karyotype of 2n = 22 = 2x = 16m + 6sm was examined for both species. The karyotypical asymmetry of 1A and 2A was found in K. chinensis and K. galioides, respectively. According to the predominant evolutionary direction of karyotype asymmetry in angiosperms, K. galioides (2A) seems slightly more evolved than K. chinensis (1A). Our finding is consistent with the hypothesis of the Old World origin of Kelloggia based on molecular study. PMID- 16705353 TI - High glucose and Nepsilon-(carboxymethyl) lysine bovine serum albumin modulate release of matrix metalloproteinases in cultured human endothelial cells. AB - BACKGROUND: Hyperglycaemia may contribute to endothelial dysfunction. Disturbances in endothelial functions include changes in the extracellular matrix underneath the cells. This may result from altered biosynthesis of matrix molecules or from modified biosynthesis and secretion of enzymes involved in the turnover of extracellular matrix. One important class of such enzymes are the matrix metalloproteinases (MMPs). AIM OF THE STUDY: The aim of this study was to investigate whether the condition of high glucose concentration relevant both to diabetes type 1 and 2 and metabolic syndrome, would affect the synthesis and release of MMPs in human umbilical cord endothelial cells (HUVEC) in vitro. METHODS: The HUVEC were isolated and cultured in vitro. The cells were exposed to medium with either low glucose (LG, 1 g/l) or high glucose (HG, 4.5 g/l) or the advanced glycation end product (AGE) N(epsilon)-(carboxymethyl) lysine bovine serum albumin (CML-BSA), at a concentration of 10 microg/ml. The HUVEC conditioned media were harvested and subjected to gelatin zymography and Western blotting. RESULTS: When HUVEC were incubated with HG or CML-BSA under serum free conditions a decreased secretion of pro MMP-2 was observed, both with gelatin zymography and Western blotting. The HUVEC also secreted MMP-9, but at lower levels, and effects of HG treatment were not significant. When HUVEC were stimulated with phorbol 12-myristate 13-acetate (PMA) secretion of pro MMP-2 was not increased, but the activation of pro MMP-2 into lower molecular forms increased, irrespective of culturing in LG, HG or CML-BSA. CONCLUSION: The HUVEC exposed to high glucose or AGE exhibit decreased secretion of MMP-2. These findings may be relevant in understanding the altered turnover of the endothelial extracellular matrix observed in the diabetic state. PMID- 16705355 TI - Spatial and temporal variations in trace metal concentrations in the cockle, Austrovenus stutchburyi from Otago, New Zealand. AB - This is the first comprehensive study of sources of variation in metal concentrations within the whole tissues of a shallow burrowing, filter-feeding intertidal clam, Austrovenus stutchburyi. Samples were collected from 12 sites in April, August, November and February in 1993-1994 in the vicinity of Otago Harbour and Peninsula, New Zealand. Total tissue trace metal concentrations (microg g(-1) dry weight) were measured in individual animals for the essential metals : Mn, Cu, Zn, Ni and the non-essential Cr using trace-metal clean acid digestion and ICP-OAES techniques. Average metal concentrations were 3-60 microg g(-1) for Cu, 40-118 microg g(-1) for Zn, 2-12 microg g(-1) for Mn, 5-35 microg g(-1) for Ni and 1-44 microg g(-1) for Cr. These levels decreased with body weight and differed amongst sites except for Cr in February (mid-summer). Highest concentrations occurred at sites close to a city (Dunedin) and within the central harbour region although the Cu, Zn, Ni and Cr concentrations did not correlate with the environmental gradient or season. At one coastal site, samples of both the blue mussel Mytilus galloprovincialis and cockles gave similar trends in trace metal levels. These results suggest that the cockle could be a useful trace metal biomonitor within NZ estuaries. PMID- 16705357 TI - NMR assignment of the phosphotyrosine binding (PTB) domain of tensin. PMID- 16705358 TI - Dynamics of transportan in bicelles is surface charge dependent. AB - In this study we investigated the dynamic behavior of the chimeric cell penetrating peptide transportan in membrane-like environments using NMR. Backbone amide 15N spin relaxation was used to investigate the dynamics in two bicelles: neutral DMPC bicelles and partly negatively charged DMPG-containing bicelles. The structure of the peptide as judged from CD and chemical shifts is similar in the two cases. Both the overall motion as well as the local dynamics is, however, different in the two types of bicelles. The overall dynamics of the peptide is significantly slower in the partly negatively charged bicelle environment, as evidenced by longer global correlation times for all measured sites. The local motion, as judged from generalized order parameters, is for all sites in the peptide more restricted when bound to negatively charged bicelles than when bound to neutral bicelles (increase in S2 is on average 0.11 +/- 0.07). The slower dynamics of transportan in charged membrane model systems cause significant line broadening in the proton NMR spectrum, which in certain cases limits the observation of 1H signals for transportan when bound to the membrane. The effect of transportan on DMPC and DHPC motion in zwitterionic bicelles was also investigated, and the motion of both components in the bicelle was found to be affected. PMID- 16705359 TI - NMR assignment of 2H, 13C and 15N labeled amino-terminal domain of apo pantothenate synthetase from E. coli. PMID- 16705360 TI - Factors affecting recurrence after incisional hernia repair. AB - Incisional hernias occur primarily as a result of high tension and inadequate healing of a previous incision, the latter of which is frequently related to infection at the surgical site. Despite recent advances in operative techniques, the recurrence rate remains unacceptably high. To evaluate the impact of different predisposing factors for the recurrence of incisional hernia, we reviewed retrospectively the medical records of 297 patients who had undergone incisional herniorrhaphy (188 tissue repairs, 109 mesh repairs) in our hospital. Demographic data (age and gender), type of repair, body mass index, hernia size, presence of chronic illnesses and wound complications were evaluated in a univariate and multivariate manner analysis. The overall recurrence rate was 30.3%, with the recurrence rate in patients who underwent tissue repair being 39.4% and that in patients following prosthetic repair 14.6%. The recurrence rate was significantly influenced by type of repair, obesity, hernia size, wound healing disorders and some chronic comorbidities. We conclude that it is necessary to become familiar with the risk factors for recurrence of incisional hernia in order to eliminate or decrease their effect on the positive outcome of incisional herniorrhaphy. PMID- 16705361 TI - Reconstruction of parasacral hernia with acellular human dermis graft. AB - Parasacral hernias are defects through the pelvic floor which occur as the result of sacral resection. These defects are often large, and are frequently the result of treatment for sacral malignancies. This report documents the case of a 71-year old woman who underwent radical coccygectomy and partial sacrectomy for a chordoma and subsequently presented 1 year later with a large parasacral hernia. The defect was repaired using an acellular human dermis graft with a gluteus maximus muscle-advancement flap overlay. This article summarizes the current literature of this challenging clinical problem, and examines the use of acellular human dermis in the repair of complex hernias. PMID- 16705362 TI - Intraperitoneal ePTFE mesh repair of parastomal hernias. AB - Parastomal hernia represents a common complication of colostomy formation. Surgical techniques such as facial repair and stoma relocation have almost been abandoned because of high recurrence rates. Extraperitoneal prosthetic mesh repair had better results but was accompanied by high rates of mesh contamination. A new technique, with intraperitoneal onlay position of expanded polytetrafluoroethylene (ePTFE) was therefore established. We report herein two cases of symptomatic large parastomal hernias treated in our department. PMID- 16705363 TI - Bilateral femoral hernia in a male cadaver with vascular variations: case report and review of the literature. AB - Femoral hernia, which is a less common occurrence than inguinal hernia, is not congenital in most cases and is uncommon in young males. It is considered to be more common in females than in males due to an enlarged femoral ring in the former. A case of bilateral femoral hernia in a 64-year-old male cadaver is described within the framework of an anatomical approach. On the right side, the protrusion of the viscus appeared as a small intestine coil, whereas on the left side the protruded viscus appeared as a pelvic colon's appendix appiplocae. On both sides, the protruded viscus was located in front of an aberrant obturator artery, which oriented from the external iliac artery and not from the internal iliac artery as should be the case. The puberal branch of the inferior epigastric artery was absent. The cadaver's medical history and his skin examination excluded an abdominal surgery. In the literature, case reports of bilateral femoral hernia appear only seldom, especially those of male patients who had not undergone inguinal hernia repair surgery. In femoral hernias more often than in other types of hernia, the protruded viscus is strangulated and undergoes a tissue necrosis. Morbidity and mortality for complicated femoral hernia is high. Knowledge of vascular variation such as presented by the cadaver under study is extremely useful to the surgeon because any iatrogenic injury of the aberrant obturator artery during a laparoscopic repair may result in dangerous hemorrhage. PMID- 16705366 TI - Predictors of reduced dose intensity in patients with early-stage breast cancer receiving adjuvant chemotherapy. AB - BACKGROUND: This retrospective study was undertaken to define risk factors for reductions in dose intensity of adjuvant chemotherapy in women with early stage breast cancer (ESBC). METHODS: A nationwide survey of 190 community oncology practices was conducted between 1998 and 2002 with data collected retrospectively on 3,707 patients treated with adjuvant chemotherapy for ESBC. End points included reductions in dose intensity, delivered relative dose intensity (RDI) <85%, and the incidence of chemotherapy dose delays >/=7 days and dose reduction >/=15%. Demographic and clinical characteristics, incidence of febrile neutropenia (FN), and patterns of use of granulocyte colony-stimulating factor (G CSF) were also assessed. RESULTS: Average RDI for all regimens was 88%, with 30% of patients receiving <85% of standard for their regimen. Seventeen percent of the reduction in average RDI was planned from the start of therapy, and 13% was unplanned. In univariate analysis, significant predictors of reduced RDI were: age >/=65 years (41%, P < 0.001), body surface area (BSA) >2 m(2) (37%, P < 0.001), negative lymph nodes (33%, P < 0.001), FN (36%, P = 0.013), and comorbidities (40%, P = 0.013), particularly renal disease (86%, P = 0.004). Dose reduction was less with prophylactic G-CSF (24%, P < 0.001). In multivariate analysis, significant independent predictors of reduced RDI included: advanced age, greater BSA, comorbidities, anthracycline-based regimens, a 28-day schedule and FN, while primary G-CSF prophylaxis was associated with a significant reduction in risk. CONCLUSION: A significant proportion of patients with potentially curable ESBC continue to experience planned and unplanned reductions in RDI. PMID- 16705367 TI - Assessment of multiple different estrogen receptor-beta antibodies for their ability to immunoprecipitate under chromatin immunoprecipitation conditions. AB - Several different antibodies to total estrogen receptor (ER)beta, ERbeta1 and ERbeta2/cx have been tested and compared for their ability to immunoprecipitate ERbeta specific isoforms under chromatin immunoprecipitation conditions (ChIP). The rabbit polyclonal antibodies AP-ERbeta1 and AP-ERbeta2/cx, specific for ERbeta1 and ERbeta2/cx isoforms, respectively, were the most efficient for ChIP. The monoclonal antibody MCA1974/PPG5/10 was also able to ChIP ERbeta1, but less efficiently than AP-ERbeta1. All other antibodies tested were not suitable for ChIP analyses although most antibodies tested immunoprecipitated the appropriate ERbeta isoforms under standard conditions. To identify antibodies that can also be used to verify in-vivo expression profiles, a comparison of the antibodies to detect ERbeta isoforms by western blotting and immunohistochemistry was also undertaken. Under the tissue processing and autostaining conditions used at the Manitoba Breast Tumor Bank 385P/GC17, MCA1974/PPG5/10, Ab288/14C8 and MCA2279S/57/3 were found to be the best for IHC of ERbeta isoforms in human breast tissue biopsy sections, while Ab14021, AP-ERbeta1 and AP-ERbeta2/cx were best for western blot detection of ERbeta isoforms. PMID- 16705370 TI - Experimental infection of big brown bats (Eptesicus fuscus) with Eurasian bat lyssaviruses Aravan, Khujand, and Irkut virus. AB - Here we describe the results of experimental infections of captive big brown bats (Eptesicus fuscus) with three newly isolated bat lyssaviruses from Eurasia (Aravan, Khujand, and Irkut viruses). Infection of E. fuscus was moderate (total, 55-75%). There was no evidence of transmission to in-contact cage mates. Incubation periods for Irkut virus infection were significantly shorter (p < 0.05) than for either Aravan or Khujand virus infections. In turn, quantification of viral RNA by TaqMan PCR suggests that the dynamics of Irkut virus infection may differ from those of Aravan/Khujand virus infection. Although infectious virus and viral RNA were detected in the brain of every rabid animal, dissemination to non-neuronal tissues was limited. Levels of viral RNA in brain of Aravan/Khujand virus-infected bats was significantly correlated with the number of other tissues positive by TaqMan PCR (p < 0.05), whereas no such relationship was observed for Irkut virus infection (where viral RNA was consistently detected in all tissues other than kidney). Infectious virus was isolated sporadically from salivary glands, and both infectious virus and viral RNA were obtained from oral swabs. The detection of viral RNA in oral swabs suggests that viral shedding in saliva occurred <5 days before the onset of clinical disease. PMID- 16705371 TI - Evaluation of the effects of a novel carbon monoxide releasing molecule (CORM-3) in an in vitro model of cardiovascular inflammation. 1. Histamine in allergy, inflammation, tissue growth and repair. PMID- 16705372 TI - Inhibition of mast cell degranulation-induced drop of blood pressure with clemastine, cromolyn and compound 48/80 pretreatment. PMID- 16705373 TI - Regulation of nerve growth factor induced histamine and arachidonic acid release from rat mast cells by cannabinoids. PMID- 16705374 TI - Analysis of IgE down regulation induced by basophil activation. Application to the diagnosis of muscle relaxant allergic hypersensitivity by flow cytometry. PMID- 16705375 TI - Use of four different flow cytometric protocols for the analysis of human basophil activation. Application to the study of the biological activity of high dilutions of histamine. PMID- 16705376 TI - Superallergens: a new mechanism of immunologic activation of human basophils and mast cells. PMID- 16705377 TI - Betahistine increases ACh release from the cortex, but not histamine release from the nucleus basalis magnocellularis of freely-moving rats. 2. Histaminergic mechanisms in the CNS. PMID- 16705378 TI - Differences in pharmacological properties of histamine H(3) receptor agonists and antagonists revealed at two human H (3) receptor isoforms. PMID- 16705379 TI - Fluorescent benzofuran histamine H(3) receptor antagonists with sub-nanomolar potency. PMID- 16705380 TI - The role of histamine and its receptors in the development of ovarian follicles in vitro. PMID- 16705381 TI - Origins of plasma amine oxidases in different mammalian species. PMID- 16705382 TI - Molecular cloning of guinea-pig diamine oxidase and histamine N methyltransferase. PMID- 16705383 TI - Histamine stimulates alphav-beta3 integrin expression of the human trophoblast through the H(1) receptor. PMID- 16705384 TI - Prostaglandin E(2) correlates with histamine production in human colorectal cancer. PMID- 16705385 TI - Histamine modulates cellular events involved in tumour invasiveness in pancreatic carcinoma cells. PMID- 16705391 TI - Surgical problems in rural Germany. PMID- 16705389 TI - Effect of antibiotic heterogeneity on the development of infections with antibiotic-resistant gram-negative organisms in a non-intensive care unit surgical ward. AB - BACKGROUND: Heterogeneous antibiotic use has been suggested to limit the emergence of resistance, but determining the optimal strategy is difficult. METHODS: We developed a new strategy, termed "periodic antibiotic monitoring and supervision" (PAMS) program in a non-ICU surgical ward. The 2-year prospective study was divided into a 1-year observation period and a 1-year PAMS period. The use of four major classes of antibiotics in empirical therapy for Gram-negative rod (GNR) infections was supervised. During the PAMS program, recommended, restricted, and off-supervised classes of antibiotics were changed every 3 months according to the usage pattern of the antibiotics in the preceding term. RESULTS: Cefepime (45.5%) and imipenem/cilastatin (39.4%) were the most common antibiotics of choice during the observation period. The use of these antibiotics decreased significantly during the PAMS period, and that of fluoroquinolones and extended spectrum penicillin/beta-lactamase inhibitor increased (4.8% vs. 21.4% and 2.4% vs. 21.4%, P<0.01 respectively). Outcome analysis demonstrated a tendency toward reduction in the incidence of resistant GNR infections (P=0.079) and that of Pseudomonas aeruginosa (P=0.053). The incidence of resistant Gram-positive core infections did not decrease. Analysis of antibiotic susceptibility to GNR revealed no significant beneficial results for any antibiotics. CONCLUSIONS: As significant changes were not observed, the PAMS program is not generally applicable and heterogeneous antibiotic use as a way of reducing infections with resistant GNR in non-ICU surgical wards was not established. PMID- 16705397 TI - Virtual CT gastrectomy by three-dimensional imaging using multidetector-row CT for laparoscopic gastrectomy. PMID- 16705398 TI - CT appearance of incidental pancreatic lipomas: a case series. AB - Pancreatic lipomas are rare benign lesions, with only 15 examples documented in the current literature. We describe five cases of intrapancreatic lipomas from a single institution, all incidentally found using computed tomography within a 14 month period. Computed tomography depicted lipomas as homogeneous, nonenhancing, well-marginated lesions with a density coefficient consistent with adipose tissue. PMID- 16705399 TI - Unusual cause of ureteral obstruction in transplant kidney. AB - Ultrasound and computed tomographic images are described in a patient who underwent renal transplantation and presented with hydronephrosis and partial ureteral obstruction secondary to herniation of the transplant ureter into a left inguinal hernia. To our knowledge, this is the first report of herniation of a transplanted ureter in the inguinal canal resulting in or exacerbating ureteral obstruction. PMID- 16705401 TI - [Invitation and observations on the 20th Surgery Congress]. PMID- 16705403 TI - A nuclear-encoded ClpP subunit of the chloroplast ATP-dependent Clp protease is essential for early development in Arabidopsis thaliana. AB - ClpP4 is a nuclear-encoded plastid protein that functions as a proteolytic subunit of the ATP-dependent Clp protease of higher plants. Given the lack of viable clpP4 knockout mutants, antisense clpP4 repression lines were prepared to study the functional importance of ClpP4 in Arabidopsis thaliana. Screening of transformants revealed viable lines with up to 90% loss of wild type levels of ClpP4 protein, while those with > 90% were severely bleached and strongly retarded in vegetative growth, failing to reach reproductive maturity. Of the viable antisense plants, repression of clpP4 expression produced a pleiotropic phenotype, of which slow growth and leaf variegation were most prominent. Chlorosis was most severe in younger leaves, with the affected regions localized around the mid-vein and exhibiting impaired chloroplast development and mesophyll cell differentiation. Chlorosis lessened during leaf expansion until all had regained the wild type appearance upon maturity. This change in phenotype correlated with the developmental expression of ClpP4 in the wild type, in which ClpP4 was less abundant in mature leaves due to post transcriptional/translational regulation. Repression of ClpP4 caused a concomitant down-regulation of other nuclear-encoded ClpP paralogs in the antisense lines, but no change in other chloroplast-localized Clp proteins. Greening of the young chlorotic antisense plants upon maturation was accelerated by increased light, either by longer photoperiod or by higher growth irradiance; conditions that both raised levels of ClpP4 in wild type leaves. In contrast, shift to low growth irradiance decreased the relative amount of ClpP4 in wild type leaves, and caused newly developed leaves of fully greened antisense lines to regain the chlorotic phenotype. PMID- 16705404 TI - Ethylene in induced conifer defense: cDNA cloning, protein expression, and cellular and subcellular localization of 1-aminocyclopropane-1-carboxylate oxidase in resin duct and phenolic parenchyma cells. AB - Members of the Pinaceae family have complex chemical defense strategies. Conifer defenses associated with specialized cell types of the bark involve constitutive and inducible accumulation of phenolic compounds in polyphenolic phloem parenchyma cells and oleoresin terpenoids in resin ducts. These defenses can protect trees against insect herbivory and fungal colonization. The phytohormone ethylene has been shown to induce the same anatomical and cellular defense responses that occur following insect feeding, mechanical wounding, or fungal inoculation in Douglas fir (Pseudotsuga menziesii) stems (Hudgins and Franceschi in Plant Physiol 135:2134-2149, 2004). However, very little is known about the genes involved in ethylene formation in conifer defense or about the temporal and spatial patterns of their protein expression. The enzyme 1-aminocyclopropane-1 carboxylate oxidase (ACO) catalyzes the final step in ethylene biosynthesis. We cloned full-length and near full-length ACO cDNAs from three conifer species, Sitka spruce (Picea sitchensis), white spruce (P. glauca), and Douglas fir, each with high similarity to Arabidopsis thaliana ACO proteins. Using an Arabidopsis anti-ACO antibody we determined that ACO is constitutively expressed in Douglas fir stem tissues and is up-regulated by mechanical wounding, consistent with the wound-induced increase of ethylene levels. Immunolocalization showed cytosolic ACO is predominantly present in specialized cell types of the wound-induced bark, specifically in epithelial cells of terpenoid-producing cortical resin ducts, in polyphenolic phloem parenchyma cells, and in ray parenchyma cells. PMID- 16705405 TI - Co-occurrence of both L-asparaginase subtypes in Arabidopsis: At3g16150 encodes a K+-dependent L-asparaginase. AB - L-asparaginases (EC 3.5.1.1) are hypothesized to play an important role in nitrogen supply to sink tissues, especially in legume-developing seeds. Two plant L-asparaginase subtypes were previously identified according to their K(+) dependence for catalytic activity. An L-asparaginase homologous to Lupinus K(+) independent enzymes with activity towards beta-aspartyl dipeptides, At5g08100, has been previously characterized as a member of the N-terminal nucleophile amidohydrolase superfamily in Arabidopsis. In this study, a K(+)-dependent L asparaginase from Arabidopsis, At3g16150, is characterized. The recombinants At3g16150 and At5g08100 share a similar subunit structure and conserved autoproteolytic pentapeptide cleavage site, commencing with the catalytic Thr nucleophile, as determined by ESI-MS. The catalytic activity of At3g16150 was enhanced approximately tenfold in the presence of K(+). At3g16150 was strictly specific for L-Asn, and had no activity towards beta-aspartyl dipeptides. At3g16150 also had an approximately 80-fold higher catalytic efficiency with L Asn relative to At5g08100. Among the beta-aspartyl dipeptides tested, At5g08100 had a preference for beta-aspartyl-His, with catalytic efficiency comparable to that with L-Asn. The phylogenetic analysis revealed that At3g16150 and At5g08100 belong to two distinct subfamilies. The transcript levels of At3g16150 and At5g08100 were highest in sink tissues, especially in flowers and siliques, early in development, as determined by quantitative RT-PCR. The overlapping spatial patterns of expression argue for a partially redundant function of the enzymes. However, the high catalytic efficiency suggests that the K(+)-dependent enzyme may metabolize L-Asn more efficiently under conditions of high metabolic demand for N. PMID- 16705406 TI - Gene conversion in human rearranged immunoglobulin genes. AB - Over the past 20 years, many DNA sequences have been published suggesting that all or part of the V(H) segment of a rearranged immunoglobulin gene may be replaced in vivo. Two different mechanisms appear to be operating. One of these is very similar to primary V(D)J recombination, involving the RAG proteins acting upon recombination signal sequences, and this has recently been proven to occur. Other sequences, many of which show partial V(H) replacements with no addition of untemplated nucleotides at the V(H)-V(H) joint, have been proposed to occur by an unusual RAG-mediated recombination with the formation of hybrid (coding-to signal) joints. These appear to occur in cells already undergoing somatic hypermutation in which, some authors are convinced, RAG genes are silenced. We recently proposed that the latter type of V(H) replacement might occur by homologous recombination initiated by the activity of AID (activation-induced cytidine deaminase), which is essential for somatic hypermutation and gene conversion. The latter has been observed in other species, but not in human Ig genes, so far. In this paper, we present a new analysis of sequences published as examples of the second type of rearrangement. This not only shows that AID recognition motifs occur in recombination regions but also that some sequences show replacement of central sections by a sequence from another gene, similar to gene conversion in the immunoglobulin genes of other species. These observations support the proposal that this type of rearrangement is likely to be AID-mediated rather than RAG-mediated and is consistent with gene conversion. PMID- 16705407 TI - LOH at 6p21.3 region and HLA class I altered phenotypes in bladder carcinomas. AB - Alterations in HLA class I antigen expression have been frequently described in different epithelial tumors and are thought to favor tumor immune escape from T lymphocyte recognition. Multiple molecular mechanisms are responsible for these altered HLA class I tumor phenotypes. Some are structural defects that produce unresponsiveness to treatment with interferons. Others include alterations in regulatory mechanisms that can be switched on by treatment of tumor cells with different cytokines. One important mechanism belonging to the first group is loss of heterozygosity (LOH) at chromosome region 6p21.3, which can lead to HLA haplotype loss. In this investigation, the frequency of LOH at 6p21 chromosome region was studied in 69 bladder carcinomas. Short tandem repeat analysis showed that 35% of cases had LOH in this chromosome region. By considering these results together with immunohistological findings previously published by our group, we identified a distribution pattern of HLA class I altered phenotypes in bladder cancer. The most frequently altered phenotype in bladder carcinomas was total loss of HLA class I expression (17 cases, 25%), followed by phenotype II associated with HLA haplotype loss (12 cases, 17.5%), and HLA allelic loss (ten cases, 14.5%). Nine cases (13%) were classified as having a compound phenotype, five cases (7%) as having HLA locus loss, and in 16 cases (23%) no alteration in HLA expression was detected. An important conclusion of this report is that a combination of different molecular and immunohistological techniques is required to precisely define which HLA alleles are lost during tumor progression and to characterize the underlying mechanisms of these losses. These studies should be performed when a cancer patient is to be included in an immunotherapy protocol that aims to stimulate different immune effector mechanisms. PMID- 16705414 TI - Rapid and quantitative extraction method for the determination of chlorophylls and carotenoids in olive oil by high-performance liquid chromatography. AB - Colour is an organoleptic characteristic of virgin olive oil and an important attribute that affects the consumer perception of quality. Chlorophylls and carotenoids are the main pigments responsible for the colour of virgin olive oil. A simple analytical method for the quantitative determination of chlorophylls and carotenoids in virgin olive oils has been developed. The pigments were isolated from small samples of oil (1.0 g) by solid-phase extraction using diol-phase cartridges (diol-SPE), and the extract was analysed by reverse-phase HPLC with diode-array UV detection. Chromatographic peak resolution, reproducibility (coefficient of variation (C.V.) <4.5%) and recovery (>98.4%) for each component were satisfactory. A comparative study of the proposed method was performed versus classical liquid-liquid extraction (LLE) with N,N'-dimethylformamide and solid-phase extraction using a C18 column (C18-SPE). While 96.4% of the pigments were recovered by LLE, only 51.3% were isolated by C18-SPE in comparison to diol SPE. Likewise, a higher alteration of pigment composition was observed when such LLE and C18-SPE procedures were used. In this sense, a higher ratio of pheophytin in comparison to that isolated by the diol-SPE procedure was achieved with both extraction procedures, indicating a greater extent of the pheophytinization reaction. Therefore, quantification of pigments from virgin olive oil by diol-SPE followed by RP-HPLC was found to be rapid, simple, required only a small amount of sample, consumed only small amounts of organic solvents, and provided high recoveries, accuracy and precision. PMID- 16705413 TI - Evaluating the interactions of vertebrate receptors with persistent pollutants and antifouling pesticides using recombinant yeast assays. AB - The development of in vitro methods for screening potentially harmful biological activities of new compounds is an extremely important way to increase not only their intrinsic environmental safety, but also the public perception of the safety standards associated with them. In this work we use two yeast systems to test the ability of different chemicals to bind and activate two vertebrate receptors which are intimately related to adverse biological effects of pollution in exposed fauna: the estrogen receptor (ER) and the aryl hydrocarbon receptor (AhR). The panel of compounds analysed here includes well-known pollutants, like PCBs, pp'-DDT and hexachlorobenzene, together with the less-known, emerging putative pollutants, such as Sea-Nine, Irgarol and diuron. Results show the ability of some of these compounds to interact with one or both receptors, provide hints about the relationship between structure and activity, and suggest mechanistic explanations for the biological activities already described in whole animal experiments. In addition, we show that AhR may have an intrinsic ligand promiscuity comparable to that of ER, a feature not fully appreciated in the past due to the technical difficulties involved with testing highly lipophilic substances in yeast-based assays. PMID- 16705418 TI - Cloning, comparative characterization of porcine SCAP gene, and identification of its two splice variants. AB - Sterol responsive element binding protein (SREBP) cleavage-activating protein (SCAP) is the key regulator of activation of SREBPs, which stimulate most enzymes in cholesterol and lipid synthesis. In order to investigate the molecular basis of lipid metabolism in the pig, a unique model for fat deposition, we isolated and characterized the porcine SCAP. The 4,096-bp full-length porcine SCAP cDNA contains an open reading frame of 3,840 bp. The predicted SCAP protein consists of 1,280 amino acids of 55-92% identity with its vertebrate counterparts. The porcine SCAP gene consists of at least 19 exons and 18 introns, which span over 13 kb of the genome. The porcine SCAP gene was mapped to chromosome 13q21-22 using a porcine-rodent somatic cell hybrid panel. Comparison of SCAP genomic structures from various species revealed intron losses in porcine, Tetraodon and fugu SCAP, and intron gains in cow and chicken SCAP. Moreover, we isolated two novel splicing SCAP variants with 193-bp (variant 2) in-frame deletion from testis and a variant with 291-bp (variant 3) in-frame deletion from liver and muscle, which may affect the function of the porcine SCAP. In conclusion, the intron gains and losses appear to have contributed to the shape of the modern SCAP family. The splice variants detected, first to be reported in any species, may be involved in the particulars of the fat metabolism in the pig. Our data lay foundation for further study of SCAP function in this species. PMID- 16705420 TI - Characterization and complementation of a Fus3/Kss1 type MAPK from Tuber borchii, TBMK. AB - Mitogen-activated protein kinases (MAPK) are used by organisms to transduce extra cellular signals from the environment in cellular events such as proliferation and differentiation. In the present study, we have characterized the first MAPK from the ectomycorrhizal fungus Tuber borchii (TBMK) which belongs to the YERK1 (yeast extra cellular regulated kinase) subfamily. TBMK is present as a single copy in the genome and the codified protein was phosphorylated during the interaction with the host plant, Tilia americana. Complementation studies showed that TBMK restores pheromone signaling in Saccharomyces cerevisiae and partially restores invasive growth of Fusarium oxysporum that lack the fmk1 gene. This suggests a protein kinase activity and its involvement in the infection processes. Hence, TBMK could play an important role during the pre-symbiotic phase of T. borchii with its host plant in the modulation of genes necessary for the establishment of symbiosis leading to the synthesis of functional ectomycorrhizae. PMID- 16705419 TI - AFLP and PCR-based markers linked to Rf3, a fertility restorer gene for S cytoplasmic male sterility in maize. AB - The Rf3 gene restores the pollen fertility disturbed by S male sterile cytoplasm. In order to develop molecular markers tightly linked to Rf3, we used amplified fragment length polymorphism (AFLP) technique with near isogenic lines (NILs) and bulk segregant analysis (BSA). A BC(1)F(1) population from a pair of NILs with different Rf3 locus was constructed and 528 primer combinations was screened. A linkage map was constructed around the Rf3 locus, which was mapped on the distal region of chromosome 2 long arm with the help of SSR marker UMC2184. The closest marker E7P6 was 0.9 cM away from Rf3. Marker E3P1, 2.4 cM from Rf3, and E12M7, 1.8 cM from Rf3, were converted into a codominant CAPS and a dominant SCAR marker, and designated as CAPSE3P1 and SCARE12M7, respectively. These markers are useful for marker-assisted selection and map-based cloning of the Rf3 gene. PMID- 16705421 TI - An experimental and theoretical study of the enantioselective deprotonation of cyclohexene oxide with isopinocampheyl-based chiral lithium amides. AB - The mechanism of the enantioselective deprotonation of cyclohexene oxide with isopinocampheyl-based chiral lithium amide was studied by quantum chemical calculations. The transition states of eight molecules were fully optimized at the ab initio HF/3-21G and density functional B3LYP/3-21G levels with Gaussian 98. The activation energies were calculated at the B3LYP/6-31+G(3df,2p)//B3LYP/3 21G level. We found the theoretical evaluation to be consistent with the experimental data. At the best case, an enantiomeric excess of up to 95% for (R) 2-scyclohexen-1-ol was achieved with (-)-N, N-diisopinocampheyl lithium amide. PMID- 16705427 TI - [Balloon kyphoplasty in the treatment of vertebral fractures]. AB - Approximately 500,000 vertebral fractures occur as a result of osteoporosis every year in Europe. One third of the patients thus affected complain of severe back pain and seek treatment. In the past, the treatment of such fractures was limited to conservative methods, such as the use of braces and analgesics and long-term immobilisation followed by physiotherapy. Since 1998 balloon kyphoplasty, a minimally invasive procedure, has also been available for their treatment. During balloon kyphoplasty a balloon system is introduced into the fractured vertebral body to achieve bitranspedicular augmentation, after which low-viscosity bone cement is injected into the vertebral body, where it sets very quickly. In general the patient can be fully mobilized 24-48 h after the procedure and in most cases the symptoms are then considerably attenuated; many patients are actually free of pain. Published studies and our own experience indicate that balloon kyphoplasty is a safe method of treating painful vertebral compression fractures sustained in various ways and that complications are rare with this procedure. PMID- 16705428 TI - [Surgical technique of balloon kyphoplasty]. PMID- 16705429 TI - [Comparison of open versus percutaneous treatment for humeral head fracture]. AB - BACKGROUND: The hypothesis of this study was that percutaneous techniques lower the risk of post-traumatic avascular necrosis. MATERIALS AND METHODS: In this retrospective study 83 patients were followed up clinically and radiologically for signs of avascular necrosis and nonunion after open or percutaneous treatment of proximal humerus fractures. Mean age was 50 years. Fractures were classified in 22 patients (26.5%) as two part, in 21 patients (25.3%) as three part, in 39 patients (47%) as four part, and in 1 patient (1.2%) as fracture dislocation (Neer classification). Fractures were treated in 12 patients (14.5%) by ORIF (open reduction and internal fixation) and in 71 patients (85.5%) by CRPF (closed reduction and percutaneous fixation). Both groups were statistically equally distributed according to fracture type (Mann-Whitney U, p=0.267) and age (One-way Annova, p=0.740). The postoperative regime did not differ between the two groups. RESULTS: Patients suffered significantly more avascular necrosis after open treatment [five patients (50%) versus eight patients (12.7%) in the percutaneous group, Mann-Whitney, p=0.004]. The risk for avascular necrosis and nonunion increased with age. Mean age of patients with avascular necrosis was 57 years, and the age of patients with nonunion was 67 years. CONCLUSION: Percutaneous treatment of humeral head fractures seems to be a reliable method for lowering the risk of avascular necrosis in young patients. PMID- 16705430 TI - [Pelvic stabilization in cases of septic instability. Triangular osteosynthesis in case of infection related vertical pelvic ring instability]. AB - The combination of transiliac screws and lumbopelvic distraction osteosynthesis is usually an appropriate procedure to treat vertical pelvic ring instabilities under the condition of full weight bearing. In this case, due to the extent of septic destruction of the dorsal portion of the iliac bone, the common triangular fixation method using conventional pedicle screws was not possible. Using the transiliac dorsoventral screw position with special long screws, we achieved high mechanical triangular stability sufficient for pelvic ring fusion despite the large bony defect. PMID- 16705431 TI - [Intertrochanteric femoral fractures in children]. AB - Based on two cases and a review of the literature, we describe a rare injury in childhood, intertrochanteric femoral fracture. Because of the insertion and traction of muscles at the proximal femur, conservative treatment is difficult. The hip muscles pull the proximal fragment into flexion, abduction and external rotation. In our opinion, surgical stabilization is the therapy of choice. We use an angular stable, locking compression plate for the stabilization of these fractures. Postoperatively, mobilization with full weight bearing is possible. PMID- 16705432 TI - Comments on the article: MW construct in fusion for neuromuscular scoliosis (Eben A. Carroll, Jeffrey Scott Shilt, Laura Jacks). PMID- 16705433 TI - The value of a pantaloon cast test in surgical decision making for chronic low back pain patients: a systematic review of the literature supplemented with a prospective cohort study. AB - The results of lumbar fusion in chronic low back pain (LBP) patients vary considerably, and there is a need for proper patient selection. Lumbosacral orthoses have been widely used to predict outcome, however, with little scientific support. The aim of the present study was to determine the value of a pantaloon cast test in selecting chronic LBP patients for lumbar fusion or conservative management. First, a systematic review of the literature was carried out in which two independent reviewers identified studies in Medline, Cochrane and Current Contents databases. Three papers met the selection criteria. In the only study with a control group, a significantly better outcome after fusion compared to conservative treatment was found in patients who reported significant pain relief while in a cast (i.e. a positive cast test). The results of lumbar fusion, however, were not significantly different for patients with a positive and those with a negative cast test. In addition to the review, a clinical cohort study of 257 LBP patients, who had been allocated to either lumbar fusion or conservative management by a temporary external transpedicular fixation trial, was performed. Prior to allocation, all had undergone a pantaloon cast test. Patients with no history of prior spine surgery and with a positive pantaloon cast test had a better outcome after lumbar fusion than those treated conservatively (P = 0.002, chi (2 )test). In patients with previous spine operations the outcomes were poor and the test was of no value. From the literature and the present patient cohort, it was concluded that only in chronic LBP patients without prior spine surgery, a pantaloon cast test with substantial pain relief suggests a favorable outcome of lumbar fusion compared to conservative management. The test has no value in patients who have had previous spine surgery. PMID- 16705434 TI - Radiation-free quantitative assessment of scoliosis: a multi center prospective study. AB - Accurate quantitative measurements of the spine are essential for deformity diagnosis and assessment of curve progression. There is much concern related to the multiple exposures to ionizing radiation associated with the Cobb method of radiographic measurement, currently the standard procedure for diagnosis and follow-up of the progression of scoliosis. In addition, the Cobb method relies on 2-D analysis of a 3-D deformity. The aim of this prospective study was to investigate the clinical value of Ortelius800 that provides a radiation-free method for scoliosis assessment in three planes (coronal, sagittal, apical), with simultaneous automatic calculation of the Cobb angle in both coronal and sagittal views. Analysis of the clinical value of the device for assessing spinal deformities was performed on patients with adolescent idiopathic scoliosis, deformity angles ranging from 10 degrees to 48 degrees. Correlation between Cobb angles measured manually on standard erect posteroanterior radiographs and those calculated by Ortelius800 showed an absolute difference between the measurements to be significantly less than +/- 5 degrees for coronal measurements and significantly less than +/- 6 degrees for sagittal measurements indicating good correlation between the two methods. The measurements from four independent sites and six independent examiners were not significantly different. We found the novel clinical tool to be reliable for following mild and moderate idiopathic curves in both coronal and sagittal planes, without exposing the patient to ionizing radiation. Considering the need for further validation of this new method, any change in treatment protocol should still be based on radiographic control. PMID- 16705435 TI - Transcriptional regulation of ZicL in the Ciona intestinalis embryo. AB - We identified 5' upstream enhancers of two Ci-ZicL genes and characterized one of them in detail. Although the genes are tandemly repeated in the genome, the transcription of each seemed to be individually regulated. The 259-bp 5' flanking sequence contained essential elements for driving a correct spatiotemporal expression. This enhancer can be divided into two distinct modules. The A module was located between nucleotide positions -259 and -205 upstream of the putative transcription start site, and was necessary for activation in A6.2 and A6.4 blastomeres at the 32-cell stage. The BM module lay between nucleotide positions 205 and -89 and was responsible for activation in B6.2 and B6.4 blastomeres at the 32-cell stage and in A-line presumptive notochord, nerve cord, and muscle lineage cells at later stages. Two putative Fox-binding sites, one located within and the other downstream of the BM module, were necessary for the latter activity. Mutation at a potential Ets-binding site, located downstream of the BM module, caused ectopic activation of the reporter gene in a-line presumptive ectoderm cells. This suggests that repression in the a-line blastomeres is necessary for correct transcriptional control of the Ci-ZicL gene. PMID- 16705436 TI - Transient receptor potential (TRP) channels as drug targets. PMID- 16705438 TI - Habitat context influences predator interference interactions and the strength of resource partitioning. AB - Despite increasing evidence that habitat structure can shape predator-prey interactions, few studies have examined the impact of habitat context on interactions among multiple predators and the consequences for combined foraging rates. We investigated the individual and combined effects of stone crabs (Menippe mercenaria) and knobbed whelks (Busycon carica) when foraging on two common bivalves, the hard clam (Mercenaria mercenaria) and the ribbed mussel (Geukensia demissa) in oyster reef and sand flat habitats. Because these species co-occur across these and other estuarine habitats of varying physical complexity, this system is ideal for examining how habitat context influences foraging rates and the generality of predator interactions. Consistent with results from previous studies, consumption rates of each predator in isolation from the other were higher in the sand flat than in the more structurally complex oyster reef habitat. However, consumption by the two predators when combined surprisingly did not differ between the two habitats. This counterintuitive result probably stems from the influence of habitat structure on predator predator interactions. In the sand-flat habitat, whelks significantly reduced their consumption of their less preferred prey when crabs were present. However, the structurally more complex oyster reef habitat appeared to reduce interference interactions among predators, such that consumption rates when the predators co occurred did not differ from predation rates when alone. In addition, both habitat context and predator-predator interactions increased resource partitioning by strengthening predator dietary selectivity. Thus, an understanding of how habitat characteristics such as physical complexity influence interactions among predators may be critical to predicting the effects of modifying predator populations on their shared prey. PMID- 16705439 TI - Spatial context influences patch residence time in foraging hierarchies. AB - Understanding responses of organisms to spatial heterogeneity in resources has emerged as a fundamentally important challenge in contemporary ecology. We examined responses of foraging herbivores to multi-scale heterogeneity in plants. We asked the question, "Is the behavior observed at coarse scales in a patch hierarchy the collective outcome of fine scale behaviors or, alternatively, does the spatial context at coarse scales entrain fine scale behavior?" To address this question we created a nested, two-level patch hierarchy. We examined the effects of the spatial context surrounding a patch on the amount of time herbivores resided in the patch. We developed a set of competing models predicting residence time as a function of the mass of plants contained in a patch and the distance between patches and examined the strength of evidence in our observations for these models. Models that included patch mass and inter patch distance as independent variables successfully predicted observed residence times (bears: r (2)=0.67-0.76 and mule deer: r (2)=0.33-0.55). Residence times of grizzly bears (Ursus arctos) and mule deer (Odocoileus hemionus) responded to the spatial context surrounding a patch. Evidence ratios of Akaike weights demonstrated that models containing effects of higher levels in the hierarchy on residence time at lower levels received up to 34 times more support in the data than models that failed to consider the higher level context for grizzly bears and up to 48 times more support for mule deer. We conclude that foraging by large herbivores is influenced by more than one level of heterogeneity in patch hierarchies and that simple empirical models offer a viable alternative to optimal foraging models for the prediction of patch residence times. PMID- 16705444 TI - A theoretical analysis of water transport through chondrocytes. AB - Because of the avascular nature of adult cartilage, nutrients and waste products are transported to and from the chondrocytes by diffusion and convection through the extracellular matrix. The convective interstitial fluid flow within and around chondrocytes is poorly understood. This theoretical study demonstrates that the incorporation of a semi-permeable membrane when modeling the chondrocyte leads to the following findings: under mechanical loading of an isolated chondrocyte the intracellular fluid pressure is on the order of tens of Pascals and the transmembrane fluid outflow, on the order of picometers per second, takes several days to subside; consequently, the chondrocyte behaves practically as an incompressible solid whenever the loading duration is on the order of minutes or hours. When embedded in its extracellular matrix (ECM), the chondrocyte response is substantially different. Mechanical loading of the tissue leads to a fluid pressure difference between intracellular and extracellular compartments on the order of tens of kilopascals and the transmembrane outflow, on the order of a nanometer per second, subsides in about 1 h. The volume of the chondrocyte decreases concomitantly with that of the ECM. The interstitial fluid flow in the extracellular matrix is directed around the cell, with peak values on the order of tens of nanometers per second. The viscous fluid shear stress acting on the cell surface is several orders of magnitude smaller than the solid matrix shear stresses resulting from the ECM deformation. These results provide new insight toward our understanding of water transport in chondrocytes. PMID- 16705445 TI - The relationship between uric acid and its oxidative product allantoin: a potential indicator for the evaluation of oxidative stress in birds. AB - Uric acid is the main nitrogenous waste product in birds but it is also known to be a potent antioxidant. Hominoid primates and birds lack the enzyme urate oxidase, which oxidizes uric acid to allantoin. Consequently, the presence of allantoin in their plasma results from non-enzymatic oxidation. In humans, the allantoin to uric acid ratio in plasma increases during oxidative stress, thus this ratio has been suggested to be an in vivo marker for oxidative stress in humans. We measured the concentrations of uric acid and allantoin in the plasma and ureteral urine of white-crowned sparrows (Zonotrichia leucophrys gambelii) at rest, immediately after 30 min of exercise in a hop/hover wheel, and after 1 h of recovery. The plasma allantoin concentration and the allantoin to uric acid ratio did not increase during exercise but we found a positive relationship between the concentrations of uric acid and allantoin in the plasma and in the ureteral urine in the three activity phases. In the plasma, the slope of the regression describing the above positive relationships was significantly higher immediately after activity. We suggest that the slope indicates the rate of uric acid oxidation and that during activity this rate increases as a result of higher production of free radicals. The present study demonstrates that allantoin is present in the plasma and in the ureteral urine of white-crowned sparrows and therefore might be useful as an indicator of oxidative stress in birds. PMID- 16705448 TI - Assessment of macular function by multifocal electroretinography following epiretinal membrane surgery with indocyanine green-assisted internal limiting membrane peeling. AB - PURPOSE: To evaluate the changes in macular function and potential retinal toxicity in epiretinal membrane (ERM) surgery with indocyanine green (ICG) assisted internal limiting membrane (ILM) peeling using multifocal electroretinography (mfERG). METHODS: Thirteen eyes of 13 patients were randomly assigned to have ERM surgery with ILM peeling using 0.5 mg/ml or 1.25 mg/ml ICG staining. Patients were examined at baseline and at 3 and 6 months postoperatively. The postoperative mfERG N1 and P1 response amplitudes and peak latencies of various ring eccentricities were assessed and compared with baseline to determine any serial changes. RESULTS: After surgery, the median best corrected visual acuity improved from 20/70 at baseline to 20/30 at 6 months postoperatively (Wilcoxon signed-ranks test, P=0.004). At 3 months postoperatively, the 0.5 mg/ml group showed no significant changes in N1 and P1 response amplitudes and peak latencies, whereas the 1.25 mg/ml ICG group showed significant reductions in N1 and P1 response amplitudes compared with baseline. No significant changes in mfERG responses were observed in either group at 6 months after surgery. CONCLUSIONS: Our mfERG findings suggested that the use of a higher concentration of ICG for ILM staining might result in transient retinal functional impairment postoperatively. The lowest possible concentration of ICG should be used intraoperatively to minimize potential retinal toxicity. PMID- 16705449 TI - Modified technique for safer indocyanine-green-assisted peeling of the internal limiting membrane during vitrectomy for macular hole repair. AB - BACKGROUND: During macular hole surgery, indocyanine green (ICG) has access to the subretinal space and can lead to toxic and phototoxic damage of the retinal pigment epithelium (RPE). To reduce its toxicity and to avoid contact between ICG and the RPE, we have developed a modified technique by using autologous whole blood. METHODS: Thirty-one eyes underwent vitrectomy for idiopathic macular hole repair. Autologous whole blood (0.1 ml) was injected into the buffered saline solution (BSS)-filled vitreous cavity over the posterior pole and aspirated with a flute cannula. A small clot remained covering the macular hole. The internal limiting membrane (ILM) was stained by using 0.05% ICG solution. The ICG was dissolved in 5% glucose to obtain an iso-osmotic solution. This ICG was injected into the BSS-filled vitreous cavity over the posterior pole and removed after 10 s. The ILM was peeled and a long-acting gas tamponade applied. Pre- and postoperative best-corrected visual acuity and optical coherence tomography (OCT) findings are reported. RESULTS: Macular hole closure was achieved in 30 of 31 eyes (97%). The mean preoperative logMAR acuity was 0.99 (range: 0.4 to 2.0). Mean postoperative logMAR acuity was 0.496 (range: 0.0 to 1.0). The average improvement in vision was 0.66 logMAR units (range: 1.5 to 0.0). No postoperative RPE alterations were observed biomicroscopically or on OCT. CONCLUSION: This surgical technique leads to favorable anatomic and functional results. ICG toxicity is reduced by modifying osmolarity, concentration and contact time and by injecting ICG under BSS. Autologous whole blood acts as a mechanical barrier and prevents ICG from entering in the subretinal space. PMID- 16705450 TI - Superficial keratectomy using microkeratome for treatment of irregular climatic droplet keratopathy (free flap). PMID- 16705451 TI - Role of signal transducer and activator of transcription 3 (STAT3) in stretch injury to bladder smooth muscle cells. AB - Excessive stretch of the bladder can lead to wall thickening including the growth of bladder smooth muscle cells (BSMC). Only three phospho-proteins (JNK, p38, and PI3K) have been previously shown to participate in stretch-induced BSMC growth. CD1 mouse bladders were hyper- or non-distended by our ex vivo bladder distention model and screened, by a commercial screening method, for phosphorylated signaling proteins. This uncovered a factor previously unexamined for its role in bladder stretch injury: signal transducer and activator of transcription 3 (STAT3). STAT3 was assessed for its role in mitogen- and stretch-induced BSMC proliferation. Proliferation was assessed by 3H-thymidine incorporation/cell counting in response to mitogenic stimulation or to stretch on silastic collagen or carboxyl-coated membranes. JAK2, upstream of STAT3, was inhibited by AG490 (2 microM). Ex vivo distention of bladders activated a discrete number of kinases, including two MAPK pathways (JNK and ERK2) and STAT3. STAT3 signaling was activated during hyperdistention of intact bladder and by stretch and mitogenic treatments of BSMC in vitro. JAK2/STAT3 inhibition by AG490 blocked mitogen- and stretch-induced BSMC proliferation. Thus, BSMC stretch responses may involve the recruitment of both growth factor and mechanically induced BSMC growth responses integrated by a common signaling pathway, STAT3. PMID- 16705453 TI - Babesiosis masquerading as recurrent immune thrombocytopenic purpura in a splenectomized patient. PMID- 16705454 TI - Acute splenic sequestration crisis in an adult with sickle beta-thalassemia. AB - Acute splenic sequestration crisis (ASSC) is a major cause of morbidity and mortality in children with sickle cell disease. Reports of ASSC in adults with sickle beta-thalassemia (S-beta(thal)) are rare and consist of isolated case reports comprising a total of seven patients, three of whom died during the crisis. We report a 22-year-old man with S-beta(thal) who developed ASSC 1 day after suffering multiple blunt trauma. Systemic inflammatory response to severe blunt trauma may have precipitated ASSC in our patient. ASSC in adults with S beta(thal) is a potentially life-threatening complication with a high risk of recurrence. Splenectomy is recommended after the first attack of ASSC in adults with S-beta(thal). PMID- 16705455 TI - Transient severe symptomatic pulmonary hypertension as onset symptom in multiple myeloma. PMID- 16705456 TI - Distinct effects of different concentrations of sodium selenite on apoptosis, cell cycle, and gene expression profile in acute promyeloytic leukemia-derived NB4 cells. AB - Selenium at a low concentration has a chemopreventive role against cancer, while at a high concentration, it exerts a direct antitumor effect. However, the mechanisms remain elusive. In this article, we discovered that Na(2)SeO(3) at 20 micromol/l concentration could significantly inhibit the proliferation of NB4 cells, affect the cell cycle distribution of cell population, and induce cellular changes characteristic of apoptotic cells, while this same compound at 2 micromol/l concentration had no such effects. The mechanisms underlying these overt differences caused by treatment of different concentrations of selenium were further investigated. cDNA microarray analysis showed that after treatment by 20 micromol/l Na(2)SeO(3), 34 genes were changed in expression, while treatment by 2 micromol/l Na(2)SeO(3) resulted in the changes of 29 genes. Nine genes were regulated in both groups, among which three showed opposite changes caused by 2 and 20 micromol/l Na(2)SeO(3). The majority of regulated genes did not coincide between the two experiment groups. In conclusion, 2 and 20 micromol/l Na(2)SeO(3) could have different effects on NB4 cells, and some genes might be involved in the underlying mechanisms. Our findings could provide basis for further uncovering the molecular mechanisms of the chemopreventive and antitumor effects of selenium and, in turn, for probing the rationality of treating leukemia with selenium. PMID- 16705457 TI - Rapid diagnosis and follow up of bladder cancer patients using urinary high molecular weight cytokeratins. AB - We have developed an office-based dot-EIA for the detection of a urinary high molecular weight cytokeratin (CK). Immunohistochemical staining and western blot based on CK1K10 monoclonal antibody were used to identify the CK. Urine of 192 patients with different types, grades, and stages of bladder tumor and 72 controls were evaluated using dot-EIA. An intense and diffuse cytoplasmic reaction was shown in bladder squamous cell carcinoma. The target epitope was identified in urine at 65, 56, and 40-kDa. The CK purified from urine showed single polypeptide at 65-kDa using SDS-PAGE and single peak at 7.4 min using capillary zone electrophoresis. The dot-EIA detected the CK with high sensitivity (97%) and specificity (94%). The CK was not detected in urine of bladder cancer patients showing response to radiotherapy. The sensitive and specific office based detection of urinary cytokeratin would be helpful in rapid diagnosis and follow up of bladder carcinoma. PMID- 16705458 TI - Spatial memory, navigation and dance behaviour in Apis mellifera. AB - Navigation and dance communication in Apis mellifera have been extensively studied on the level of sensory processing, but the structure and content of the spatial memory underlying such phenomena have yet to be addressed. Here we survey new findings indicating that the memory used by bees to navigate within the range of their orientation flights is much more complex than hitherto thought. It appears to allow them to decide between at least two goals in the field, and to steer towards them over considerable distances. Two models concerning the structure of bees' spatial memory are developed from new empirical evidence. The first one relies on the integration of at least two flight vectors, while the second assumes the existence of a 'functional' map based on the information available on-site. These findings also raise questions about the process of encoding and decoding information in the context of the waggle dance. We review published data and recent evidence indicating that memories of topographical features might also be involved in dance communication, and point out what needs to be addressed to elucidate the corresponding memory demands. The flight paths of recruited bees can now be traced by means of radar techniques, and thus tools are available to tackle these questions. PMID- 16705459 TI - Comparison of single-pool and equilibrated Kt/V values for pediatric hemodialysis prescription management: analysis from the Centers for Medicare & Medicaid Services Clinical Performance Measures Project. AB - Current formulas that estimate the delivered dose of hemodialysis rely upon pre- and post-treatment blood urea nitrogen (BUN) concentrations for calculation. Single-pool kinetic modeling (spKt/V) uses a convenient 30-s post-dialysis BUN sample but does not take urea rebound into account. Double-pool modeling (eKt/V) uses an equilibrated BUN (eqBUN) and is the best reflection of the true urea mass removed by hemodialysis but is inconvenient for patients and costly to the dialysis unit to wait to obtain an eqBUN sample. We compared simple spKt/V and eKt/V estimation formulas using data obtained from the Centers for Medicare & Medicaid Services (CMS) End Stage Renal Disease (ESRD) Clinical Performance Measures (CPM) Project to determine how frequently these two results would lead to different prescription management. We set an expected difference Kt/V (spKt/V eKt/V) of 0.20 based on results of the Hemodialysis (HEMO) Study; 1,513 paired spKt/V and estimated eKt/V results were available for comparison. For patients with an arteriovenous fistula (AVF) or arteriovenous graft (AVG) (n=720), mean spKt/V and estimated eKt/V were 1.62+/-0.30 and 1.37+/-0.26, respectively. For patients with a catheter (n=793), mean spKt/V and estimated eKt/V were 1.53+/ 0.32 and 1.33+/-0.29, respectively. Examination of the different spKt/V and estimated eKt/V pairings revealed a greater adequacy discordance rate between a 0.20 difference in spKt/V and estimated eKt/V at higher Kt/V values, but Kt/V discordance rates only varied from 0.3 to 5.5% depending on the paired Kt/V values used. PMID- 16705460 TI - [Nocardia farcinica infection of a vascular prosthesis]. AB - A 61-year-old woman presented with purple-red subcutaneous nodules on her right leg. She had experienced fever up to 40 degrees C for the past 10 days and felt generally weak over the last two months. Four months earlier, a vascular graft had been implanted in her right femoral artery. Based on the diagnosis of skin infection due to implantation of an infected prosthesis, she was hospitalized and treated with an antibiotic regime. During the initial antibiotic treatment, the symptoms deteriorated, and she developed joint, hepatic and CNS abscesses. Finally, a microbiologic culture with an extended incubation time revealed the diagnosis of an infection with the slowly growing bacterium Nocardia ssp. Hereupon the patient was treated purposefully with Meropenem over six months. This treatment resulted in complete recovery. PMID- 16705461 TI - [Dermatitis artefacta presenting as photodermatosis]. AB - Dermatitis artefacta is a form of a self-injury due to psychiatric disorders or internal conflicts. Delayed diagnosis often leads to unnecessary treatments. A 17 year old girl was referred with a putative photodermatosis presenting with erosions on an erythematous base on the face and forearms. The unusual rapid onset of new lesions following phototesting and in particular a reaction induced after a simulated light exposure substantiated the diagnosis of dermatitis artefacta. Faced with the diagnosed the patient admitted she had induced the lesions and was referred for psychiatric care. PMID- 16705462 TI - [Juvenile verrucae planae: treatment with imiquimod 5% cream]. AB - A 6 year old girl presented with flat-topped, skin-colored, partly grouped papules with slightly roughened surface on the left side of the forehead and around the left orbit. In an off-label use imiquimod 5% cream was applied twice a week before bedtime and was rinsed off in the morning. After four weeks of treatment the lesions had vanished completely. We suggest imiquimod 5% cream as a treatment option for juvenile plane warts even in children and in the face. PMID- 16705463 TI - The prevalence of sexual dysfunction and associated risk factors in women with chronic pelvic pain: a cross-sectional study. AB - The aims of the study were to determine the prevalence of sexual dysfunction, its subtypes and associated risk factors in women with chronic pelvic pain (CPP) as compared to a general female population. We evaluated 112 women (mean age 34.73 +/- 8.07; age range 18-50) complaining of CPP with a comprehensive history including female sexual function index (FSFI) and several general assessment questions (GAQs), a complete physical examination and routine laboratory tests. A group of 108 healthy women (mean age 33.28 +/- 7.95; age range 19-52) without CPP were enrolled as cross-sectional controls. According to the general population, the incidence of female sexual dysfunction (FSD) was 67.8% in women with CPP and 32.2% in women without CPP (P < 0.0001). Among 112 CPP patients, 78 (69.6%) of them had FSD and 34 (30.4%) patients did not have FSD in the study (P < 0.0001). In that 78 patients, 42 patients (53.8%) had hypoactive sexual desire disorder, 26 patients (33.3%) had sexual arousal disorder, 17 patients (21.7%) had orgasmic disorder and finally 58 patients (74.3%) had sexual pain disorder. The FSFI scores in both groups were as follows: (patients vs. controls; median value; P value, respectively): desire: 3.31 versus 3.98 (P < 0.0001); arousal: 3.58 versus 4.35 (P < 0.0001); lubrication: 4.20 versus 4.88 (P < 0.0001); orgasm: 3.70 versus 4.48 (P < 0.0001); sexual satisfaction: 3.80 versus 4.64 (P < 0.0001); sexual pain: 2.75 versus 4.98 (P < 0.0001) and total FSFI score: 21.35 versus 27.29 (P < 0.0001). The prevalence of FSD was higher in women with CPP than in a general healthy population not complaining of CPP. Investigation of female sexuality was essential for these patients. PMID- 16705464 TI - Expressed sequence tag analysis of adult Clonorchis sinensis, the Chinese liver fluke. AB - Expressed sequence tag (EST) pools represent partial profiles of the gene expressions of organisms. In an effort to construct a Clonorchis sinensis EST pool, 2,387 ESTs were collected from an adult C. sinensis cDNA library and assembled into 1,573 clusters. Of these clusters, 1,225 ESTs (51%) were singletons and 348 clusters consisted of more than two ESTs. There were 848 clusters (54%) that shared significant identity with previously reported proteins, and of these, 401 clusters were categorized into 11 major functional protein classes. Three cDNA clones of fructose-1,6-bisphosphate (FBP) aldolase were selected from the C. sinensis EST pool and analyzed for phylogenic clustering. FBP clones encoded a complete polypeptide, which shared significant identity to those of vertebrate and invertebrate animals and clustered with those of trematodes. We believe that the EST pool described can be confidently used as a platform in multigene researches on C. sinensis gene expression. PMID- 16705465 TI - The contribution of individual and pairwise combinations of SNPs in the APOA1 and APOC3 genes to interindividual HDL-C variability. AB - Apolipoproteins (apo) A-I and C-III are components of high-density lipoprotein cholesterol (HDL-C), a quantitative trait negatively correlated with risk of cardiovascular disease (CVD). We analyzed the contribution of individual and pairwise combinations of single nucleotide polymorphisms (SNPs) in the APOA1/APOC3 genes to HDL-C variability to evaluate (1) consistency of published single-SNP studies with our single-SNP analyses; (2) consistency of single-SNP and two-SNP phenotype-genotype relationships across race-, gender-, and geographical location-dependent contexts; and (3) the contribution of single SNPs and pairs of SNPs to variability beyond that explained by plasma apo A-I concentration. We analyzed 45 SNPs in 3,831 young African-American (N=1,858) and European-American (N=1,973) females and males ascertained by the Coronary Artery Risk Development in Young Adults (CARDIA) study. We found three SNPs that significantly impact HDL-C variability in both the literature and the CARDIA sample. Single-SNP analyses identified only one of five significant HDL-C SNP genotype relationships in the CARDIA study that was consistent across all race-, gender-, and geographical location-dependent contexts. The other four were consistent across geographical locations for a particular race-gender context. The portion of total phenotypic variance explained by single-SNP genotypes and genotypes defined by pairs of SNPs was less than 3%, an amount that is miniscule compared to the contribution explained by variability in plasma apo A-I concentration. Our findings illustrate the impact of context-dependence on SNP selection for prediction of CVD risk factor variability. PMID- 16705466 TI - Compliance with recommended immunizations in adolescents. AB - INTRODUCTION: Little is known about the completeness and timely administration of recommended standard immunizations in Germany. The goal of this study was to determine compliance with official standard immunization recommendations in adolescents attending secondary schools in the city of Erlangen, Germany. METHODS: Adolescents who were attending 5th grade (at approximately 11 years of age), 8th grade (14 years), or 10th and 11th grade (16-17 years) classes at any of the 13 of 14 schools that had agreed to participate were eligible to be enrolled. RESULTS: While coverage for the primary series of diphtheria, tetanus and poliomyelitis immunizations was satisfactory (98%), coverage for measles mumps-rubella immunizations (dose 1: 89-96%; dose 2: 60-76%) and hepatitis B (doses 1-3: 61%) was suboptimal. Of note, 39% of students had not received any immunization against pertussis. Completion of immunization series generally was significantly delayed. Furthermore, rates for recommended booster doses in adolescence were disappointingly low with 21% for tetanus component vaccines and <10% for the fifth dose of pertussis. CONCLUSIONS: Significant immunization gaps for all recommended standard immunizations in adolescents were detected. This puts individuals at risk for serious vaccine-preventable diseases, contributes to suboptimal herd immunity in the population under study leaving the potential for future epidemics, and impedes national and international targets of disease reduction or elimination. PMID- 16705467 TI - Gastrointestinal oxalic acid absorption in calcium-treated rats. AB - We studied whether urinary oxalate excretion after an acute oral load of oxalic acid is influenced by concomitant administration of calcium in rats. Male Wistar rats weighing approximately 180 g were divided into six groups of five animals each. After inducing anesthesia, the animals were orally (via a gastrostomy) given 110 micromol of oxalic acid along with 0, 27.5, 55, 110, or 220 micromol of calcium (0, 27.5, 55, 110, or 220 micromol Ca group, respectively). Saline was given to the control group instead of oxalic acid. Urine specimens were collected before administration and then at hourly intervals up to 5 h afterward. Urinary oxalate and citrate levels were measured by capillary electrophoresis, while urinary calcium, magnesium, and phosphorus levels were measured by ICP spectrophotometry. Urinary oxalate excretion peaked at 1 h after administration and was higher in the 0, 27.5, and 55 micromol Ca groups than in the control group. The urinary recovery of oxalate in these groups was 10-15%, while the recovery rate was less than 3% in other groups. Urinary Ca excretion showed no significant changes, either over time or between groups. Free oxalic acid is absorbed more readily from the gastrointestinal tract than calcium oxalate, while simultaneous administration of calcium appears to block intestinal oxalic acid absorption in a dose-dependent manner. PMID- 16705468 TI - Beta1-Adrenergic receptor antagonism abrogates cardioprotective effects of intermittent hypoxia. AB - Adaptation to hypoxia lessens myocardial ischemic injury. This study tested whether hypoxia-induced beta-adrenergic activity mobilizes mechanisms that protect myocardium during subsequent ischemia and reperfusion. Dogs were intermittent hypoxia conditioned (IHC) by a 20 days program of 5-8 daily, 5-10 min cycles of normobaric hypoxia (FIO2 = 9.5-10%), or sham conditioned with normoxic air, and metoprolol (beta1-adrenoceptor antagonist) was administered throughout the IHC program. Twenty-four hours after the last IHC session, the left anterior descending coronary artery (LAD) was occluded for 60 min, and then reperfused for 5 h. Area at risk (AAR) and infarct size (IS) were measured. IHC lowered IS/AAR from 38+/-6% in sham-conditioned dogs to 1.1+/-0.3%, and eliminated ventricular tachycardia (VT) and fibrillation (VF) that occurred in 14 of 17 non-conditioned dogs. Metoprolol blunted IHC-evoked cardioprotection (IS/AAR=27+/-3%), and VT and/or VF occurred in 5 of 6 dogs. Metoprolol did not exacerbate ischemic injury in sham-conditioned dogs (IS/AAR=38+/-2%). Neither IHC nor metoprolol affected hematocrit or LAD collateral blood flow. A single IHC session failed to protect ischemic myocardium (IS/AAR = 36+/-8%), and protection was incomplete after 10 days of IHC (IS/AAR = 13+/-5%), suggesting that de novo protein synthesis was required for protection. Thus, episodic beta1-adrenergic activation during IHC evokes progressive development of powerful resistance to myocardial ischemia. PMID- 16705469 TI - Brief reoxygenation episodes during chronic hypoxia enhance posthypoxic recovery of LV function: role of mitogen-activated protein kinase signaling pathways. AB - Children with congenital cyanotic heart defects have worse outcomes after surgical repair of their heart defects compared with noncyanotic ones. Institution of extracorporeal circulation in these children exposes the cyanotic heart to reoxygenation injury. Mitogen-activated protein kinase (MAPK) signaling cascades are major regulators of cardiomyocyte function in acute hypoxia and reoxygenation. However, their roles in chronic hypoxia are incompletely understood. We determined myocardial activation of the three major MAPKs, c-Jun NH2-terminal kinase (JNK), extracellular signal-regulated kinase-1/2 (ERK1/2), and p38-MAPK in adult rats exposed to hypoxia (FIO2=0.10) for varying periods of time. Myocardial function was analyzed in isolated perfused hearts. Acute hypoxia stimulated JNK and p38-MAPK activation. Chronic hypoxia (2 weeks) was associated with increased p38-MAPK (but not JNK) activation, increased apoptosis, and impaired posthypoxic recovery of LV function. Brief normoxic episodes (1 h/day) during chronic hypoxia abolished p38-MAPK activation, stimulated MEK-ERK1/2 activation modestly, and restored posthypoxic LV function. In vivo p38-MAPK inhibition by SB203580 or SB202190 in chronically hypoxic rats restored posthypoxic LV function. These results indicate that sustained hypoxemia maintains p38-MAPK in a chronically activated state that predisposes to myocardial impairment upon reoxygenation. Brief normoxic episodes during chronic hypoxia prevent p38-MAPK activation and restore posthypoxic recovery of myocardial function. PMID- 16705470 TI - Expression of nitric oxide related enzymes in coronary heart disease. AB - Enzymes involved in the metabolism nitric oxide (NO) and reactive oxygen species (ROS) may play a role for the decreased availability of NO in atherosclerosis. We, therefore, hypothesized that the pattern of gene expression of these enzymes is altered in atherosclerosis. Myocardial tissue from patients with coronary heart disease (CHD) or without CHD (control group) was investigated. The level of enzymes related to NO/ROS metabolism was determined both at mRNA level and protein level by rt-PCR, real-time PCR, and western blot. The expression of NOS1 3 (synthesis of NO), arginase1 (reduction of L-arginine), p22phox (active subunit of NADPH oxidase), GTPCH (rate limiting enzyme for tetrahydrobiopterin), SOD1-3 (scavengers of superoxide anions), PRTMT1-3, and DDAH2 (involved in the metabolism of ADMA) was determined. All enzymes were found to be expressed in human myocardium. NOS isoforms were decreased in CHD in protein level, but only the downregulation of NOS3 expression reached statistical significance. The expression of PRMT1 and PRMT3 was increased. In addition, the expression of DDAH2 was reduced, both theoretically leading to an increase of ADMA concentration. SOD3 was downregulated in tissue from patients with CHD. Taken together, in myocardial tissue from patients with atherosclerosis, the expression of genes increasing ADMA levels is enhanced in contrast to a reduced expression of genes promoting NO synthesis. These results may contribute to the explanation of increased oxidative stress in atherosclerosis on the level of gene expression. PMID- 16705471 TI - Transcoronary delivery of bone marrow cells to the infarcted murine myocardium: feasibility, cellular kinetics, and improvement in cardiac function. AB - Efficient strategies for labelling and delivery of bone marrow derived stem cells (BMCs) are required to elucidate the cellular kinetics and therapeutic effects after BMC transfer for myocardial infarction (MI). Lineage negative (lin-) BMCs, labelled ex vivo in a simple procedure with the cell tracker dye tetramethyl rhodamine (TAMRA), were reliably detected by fluorescence microscopy with higher specificity than retroviral enhanced green fluorescence protein (EGFP) marking and detection. Only few cells entered the ischemic myocardium after intravenous (i.v.) application, but this number increased more than 18-fold after transcoronary delivery. Time course and kinetic analysis over 12 h revealed that myocardial colonization seems to be a biphasic process of first order decay with different elimination half-lives. Most cells are eliminated rapidly during the first 2 h (t1/2 40 min), but the remaining cells are retained significantly longer in the ischemic heart (t1/2 5.2 h). In contrast, BMC colonization of the spleen increased rather in a linear fashion. Although transcoronary BMC transfusion did not alter infarct size, it increased capillary density in the infarct border zone and improved LV function 4 weeks after MI. In conclusion, BMCs delivered by transcoronary injection increase capillary density and improve LV function after MI although homing to the ischemic heart is only transient. PMID- 16705474 TI - Tau haplotype affects CSF Tau levels in frontotemporal dementia: implication for diagnostic purposes. PMID- 16705475 TI - Monophasic demyelinating disease of the central nervous system associated with Hepatitis A infection. PMID- 16705476 TI - Mislocalization of tactile stimulation in patients with complex regional pain syndrome. AB - Complex-Regional-Pain-Syndromes (CRPS) are characterized by sensory,motor and autonomic dysfunctions. Patterns of sensory symptoms suggest changes within the central nervous system (CNS). Recently, we could show substantial reorganization of somatotopic maps within the central nervous system of patients with CRPS using functional imaging techniques (Maihofner et al. Neurology, 2003). These changes were predicted by CRPS pain and mechanical hyperalgesia. In the present study we looked for potential psychophysical correlates of cortical reorganization in CRPS. Sequential pneumatic non-noxious tactile stimulation was performed at digits 1 and 5 in 24 patients with CRPS of the upper extremities. Both the unaffected and affected side were examined. Patients were interviewed for tactile induced sensations. The occurrence of mislocalizations was correlated with a detailed psychophysical examination in which sensory, motor and autonomic symptoms were assessed. Eight patients (30 %) reported tactile mislocalizations, which were felt in the affected hand. In four cases the referred sensations spread into other nerve territories (ulnar/median nerve). Presence of mechanical hyperalgesia significantly predicted the occurrence of mislocalizations. In contrast, in a healthy control group, no mislocalizations were found. Thus, our results further support the concept of pain-induced reorganization in the somatosensory system of CRPS patients. PMID- 16705477 TI - Right ventricular diastolic abnormalities in rheumatoid arthritis and its relationship with left ventricular and pulmonary involvement. A tissue Doppler echocardiographic study. AB - OBJECTIVES: To investigate right ventricular diastolic function in rheumatoid arthritis (RA) and its relationship with left ventricular and pulmonary involvement. METHODS: Thirty-five RA patients and 30 healthy subjects were submitted to conventional Doppler (CE) and tissue Doppler echocardiography (TDE) to assess left and right systolic and diastolic function and to estimate maximal arterial systolic pulmonary pressure (PAP). To detect pulmonary involvement, pulmonary function tests and high-resolution computed tomography (HRCT) scans were performed in all RA patients. RESULTS: An abnormal RV filling, as expressed byan inverted tricuspid (Tr.) E/A ratio, was detected in 12 (34%) of the 35 RA patients and in 2 (7%) of the 30 controls (P<0.004). If compared to CE findings, prevalence of RV diastolic abnormalities were found higher in patients with RA by TDE (RV annulus Em/Am ratio <1 (in 31 (89%) of 35 patients) (P = 0.002). Twenty two (63%) of 35 patients had abnormal HRCT findings. Pulmonary involvement with pulmonary hypertension (PHT) (36+/-5 mmHg) was detected in 10 (29%) of 35 RA. In this group, increase of RV annulus and basal Am wave, decrease of Tr. E/A ratio and RV annulus Em/Am ratio were statistically significant compared to RA (12 (34%) of 35) patients with pulmonary involvement who had normal PAP (19+/-5 mmHg), (P = 0.014, P = 0.006, P = 0.015, P = 0.049, respectively). CONCLUSIONS: This study points out an impaired RV filling in a significant part of RA patients without overt heart failure. Impairment of RV diastolic function may be a predictor of subclinic myocardial and pulmonary involvement in patients with RA. PMID- 16705478 TI - Diagnostic features of cardiac hemangioma on cardiovascular magnetic resonance, a case report. AB - Cardiac hemangiomas are benign cardiac tumors that account for 5-10% of all benign tumors of the heart (Grebenc ML, Rosado de Christenson ML, Burke AP, Green CE, Galvin JR. Radiographics 2000; 20(4): 1073-1103). They occur in any cardiac location, including the pericardium (Brodwater B, Erasmus J, McAdams HP, Dodd L. J Comput Assist Tomogr 1996; 20(6): 954-956). Magnetic resonance imaging (MRI) has an excellent contrast resolution and multiplanar capability to allow optimal evaluation of myocardial infiltration, pericardial involvement and/or extracardiac extension (Brown JI, Barakos JA, Higgins CB. J Thorac Imaging 1989; 4(2): 58-64). This is a case report of cardiac hemangioma involving the ventricular septum with radiological and pathological correlation. It illustrates the capability of the MRI to non-invasively detect histological and flow characteristics of the tumor. PMID- 16705479 TI - A randomized clinical trial of two telephone-delivered, mental health interventions for HIV-infected persons in rural areas of the United States. AB - Through December 2003, more than 55,300 persons were living in small towns and rural areas of the United States at the time of their AIDS diagnosis. Many HIV infected rural persons experience elevated levels of depression, suicidal ideation, and anxiety. This research tested if two telephone-delivered, mental health interventions could facilitate the adjustment efforts of persons living with HIV/AIDS in rural areas of the United States. Participants (N=299) were recruited through AIDS service organizations in 13 states and assigned to a Usual Care Condition (n=107), an 8-session Information Support Group Intervention (n=84), or an 8-session Coping Improvement Group Intervention (n=108). Participants completed self-administered surveys at pre-intervention, post intervention, and 4- and 8-month follow-ups. ANCOVA revealed that no treatment condition produced reductions in the main outcome measures of depressive and psychological symptoms; however, Information Support participants received significantly more support from friends at 4- and 8-month follow-ups and reported fewer barriers to health care and social services at 4-month follow-up compared to participants in the other two conditions. Telephone-delivered, information support groups have potential to increase perceptions of support and reduce barriers to health care and social services in this population, but the practical significance of these intervention-related changes is limited. PMID- 16705480 TI - Development of the new group of indole-derived neuroprotective drugs affecting oxidative stress. AB - 1. The role of oxidative stress, and accordingly uncontrolled reactive oxygen species generation/action, have been widely documented in a number of different neuronal pathologies. However, the concept of pharmacological interventions in prevention and therapy of oxidative stress-related diseases has not found adequate application in clinical practice. This may be due to the insufficient efficacy of drugs available, their unsuitable pharmacokinetics, side effects, toxicity, etc. 2. Based on stobadine, (--)-cis-2,8-dimethyl-2,3,4,4a,5,9b hexahydro-1H-pyrido[4,3-b]indole, a well-known antioxidant, free radical scavenger, and neuroprotectant, it was attempted to develop new stobadine derivatives with improved pharmacodynamic and toxicity profiles, on applying molecular design, synthesis and adequate tests. Stobadine molecule was modified mostly by electron donating substitution on the benzene ring and by alkoxycarbonyl substitution at N-2 position. A total of >70 derivatives were prepared. 3. In a mice model of head trauma, some of the new stobadine derivatives administered i.v. immediately after the trauma, significantly improved sensomotoric outcome in the animals assessed 1 h later. Accordingly, decrease in brain edema was proved histologically as well as by brain wet weight assessment. 4. Putative neuroprotective action of the compounds was confirmed on rat hippocampal slices exposed to reversible 6 min hypoxia/low glucose by analysis of synaptic transmission in CA1 region neurons. Irreversible impairment of neurotransmission resulting from the hypoxia was significantly reduced by the presence of SMe1EC2, one of the new compounds, in concentration range 0.03 10.0x10(-6) mol l(-1). Both the neuroprotective and antioxidant effect of the compound closely resembled those of stobadine, melatonin, 21-aminosteroids, alpha phenyl-tert-butylnitrone and others, all well-established antioxidants, except the range of effective concentrations was by 1-2 orders lower in SMe1EC2. 5. A remarkable antioxidant efficacy was observed in the new compounds in rat brain homogenates exposed to iron/ascorbate system by protection of lipids and creatine kinase against the oxidative impairment. A link between the neuroprotective and antioxidant/ scavenger properties in the compounds can be assumed. 6. Acute toxicity of some of the new pyridoindoles was diminished compared to stobadine. That might be due to the virtually full elimination of stobadine's undesired alpha (1)-adrenolytic activity attained by appropriate modifications of its molecule. 7. The new pyridoindoles extend the range of available neuroprotectants interfering with oxidative stress in neuronal tissue. PMID- 16705482 TI - Intra- and extraneuronal changes of immunofluorescence staining for TNF-alpha and TNFR1 in the dorsal root ganglia of rat peripheral neuropathic pain models. AB - 1. Several lines of evidence suggest that cytokines and their receptors are initiators of changes in the activity of dorsal root ganglia (DRG) neurons, but their cellular distribution is still very limited or controversial. Therefore, the goal of present study was to investigate immunohistochemical distribution of TNF-alpha and TNF receptor-1 (TNFR1) proteins in the rat DRG following three types of nerve injury. 2. The unilateral sciatic and spinal nerve ligation as well as the sciatic nerve transection were used to induce changes in the distribution of TNF-alpha and TNFR1 proteins. The TNF-alpha and TNFR1 immunofluorescence was assessed in the L4-L5 DRG affected by nerve injury for 1 and 2 weeks, and compared with the contralateral ones and those removed from naive or sham-operated rats. A part of the sections was incubated for simultaneous immunostaining for TNF-alpha and ED-1. The immunofluorescence brightness was measured by image analysis system (LUCIA-G v4.21) to quantify immunostaining for TNF-alpha and TNFR1 in the naive, ipsi- and contralateral DRG following nerve injury. 3. The ipsilateral L4-L5 DRG and their contralateral counterparts of the rats operated for nerve injury displayed an increased immunofluorescence (IF) for TNF-alpha and TNFR1 when compared with DRG harvested from naive or sham-operated rats. The TNFalpha IF was increased bilaterally in the satellite glial cells (SGC) and contralaterally in the neuronal nuclei following sciatic and spinal nerve ligature. The neuronal bodies and their SGC exhibited bilaterally enhanced IF for TNF-alpha after sciatic nerve transection for 1 and 2 weeks. In addition, the affected DRG were invaded by ED-1 positive macrophages which displayed simultaneously TNFalpha IF. The ED-1 positive macrophages were frequently located near the neuronal bodies to occupy a position of the satellites. 4. The sciatic and spinal nerve ligature resulted in an increased TNFR1 IF in the neuronal bodies of both ipsi- and contralateral DRG. The sciatic nerve ligature for 1 week induced a rise in TNFR1 IF in the contralateral DRG neurons and their SGC to a higher level than in the ipsilateral ones. In contrast, the sciatic nerve ligature for 2 weeks caused a similar increase of TNFR1 IF in the neurons and their SGC of both ipsi- and contralateral DRG. The spinal nerve ligature or sciatic nerve transection resulted in an increased TNFR1 IF located at the surface of the ipsilateral DRG neurons, but dispersed IF in the contralateral ones. In addition, the SGC of the contralateral in contrast to ipsilateral DRG displayed a higher TNFR1 IF. 5. Our results suggest more sources of TNF-alpha protein in the ipsilateral and contralateral DRG following unilateral nerve injury including macrophages, SGC and primary sensory neurons. In addition, the SGC and macrophages, which became to be satellites, are well positioned to regulate activity of the DRG neurons by production of TNF-alpha molecules. Moreover, the different cellular distribution of TNFR1 in the ipsi- and contralateral DRG may reflect different pathways by which TNF-alpha effect on the primary sensory neurons can be mediated following nerve injury. PMID- 16705481 TI - Feast or famine: the sympathetic nervous system response to nutrient intake. AB - : 1. The use of tritiated norepinephrine (NE) to measure the turnover rate of NE in sympathetically innervated organs was pioneered in the laboratory of Julius Axelrod. This technique provides an organ specific assessment of sympathetic activity, integrated over a 24 h period, in free living laboratory animals. As such it has proved useful in estimating changes in sympathetic outflow in different physiologic and patho-physiologic states. 2. Studies employing NE turnover techniques in laboratory rodents have demonstrated conclusively that fasting suppresses and overfeeding stimulates the sympathetic nervous system (SNS). These changes in sympathetic activity also occur in humans. 3. Diet induced changes in SNS activity are regulated by insulin-mediated glucose uptake and metabolism in central neurons sensitive to insulin and located anatomically in the ventro-medial hypothalamus. The regulation is imposed by descending inhibition of tonically active sympathetic brainstem centers. 4. Diet-induced changes in SNS activity mediate changes in energy production known as dietary thermogenesis. The capacity for dietary thermogenesis serves as a potential buffer against weight gain.5. Insulin stimulated SNS activity contributes to obesity-related hypertension. The insulin resistance of obesity, and consequent hyperinsulinemia, drives sympathetically mediated thermogenesis, restoring energy balance at the expense of SNS over activity. The association of obesity and hypertension, therefore, may be the unintended consequence of mechanisms recruited in the obese to limit further weight gain. PMID- 16705484 TI - Mood states associated with transitory physical symptoms among breast and ovarian cancer survivors. AB - This study used electronic diaries to examine patterns of mood and physical symptoms within and across days in two independent samples of cancer patients. Twenty-three breast cancer survivors (post-treatment) and 33 ovarian cancer survivors (on chemotherapy) recorded mood and physical symptoms 4 times daily for 7 consecutive days. A series of repeated-measures multilevel models using SAS Proc Mixed were calculated to estimate the degree to which physical symptoms (e.g., pain, fatigue, and nausea) were associated with participants' moods. Across days, mood vectors with a pleasantness component (i.e., happy-sad and calm anxious) and mood vectors with an arousal component (i.e., active-passive and peppy-tired) were significantly associated with physical symptom severity. Specifically, breast cancer survivors with greater fatigue and pain reported more negative moods (eta2 < or = 0.33). Ovarian cancer survivors with greater fatigue (eta2 < or = 0.35), pain (eta2 < or = 0.04), and nausea (eta2 < or = 0.04) also reported more negative moods. Diurnal analyses showed that happy-sad (eta2 < or = 0.16), active-passive (eta2 < or = 0.27), and peppy-tired moods (eta2 < or = 0.33) were significantly negatively associated with fatigue at each of the four daily assessment times in both samples. Although correlational, our findings are consistent with previous studies suggesting that variations in both pleasant and aroused mood covary with changes in real-time physical symptom reports. PMID- 16705486 TI - Efficacy, safety and tolerability of a new 10% liquid intravenous immune globulin [IGIV 10%] in patients with primary immunodeficiency. AB - The present clinical study was designed to evaluate the efficacy, pharmacokinetics and safety of a new 10% liquid intravenous immune globulin in patients with primary immunodeficiency diseases. Sixty-one adults and children with primary immuno-deficiency diseases received doses of 300-600 mg/kg body weight every 21-28 days for 12 months. No validated acute serious bacterial infections were reported. The 95% confidence interval for the annualized rate of acute serious bacterial infections (primary endpoint) was 0-0.060. A total of four predefined validated other bacterial infections commonly occurring in primary immunodeficiency disease subjects were observed; none were serious, severe or resulted in hospitalization. The median elimination half-life of IgG was 35 days. Median total IgG trough levels varied from 9.6 to 11.2 g/L. Temporally associated adverse experiences were determined for 72 h after each infusion and the most common adverse experience was headache, which was associated with 6.9% of infusions. The study met the primary endpoint for efficacy and demonstrated excellent tolerability of the new 10% liquid intravenous imunoglobulin preparation. PMID- 16705488 TI - Interspecific pheromone cross-attraction among soybean bugs (Heteroptera): does Piezodorus hybneri (Pentatomidae) utilize the pheromone of Riptortus clavatus (Alydidae) as a kairomone? AB - The chemical and ecological function of cross-attraction of Piezodorus hybneri (Pentatomidae) to the Riptortus clavatus (Alydidae) pheromone (a mixture of three components) was studied. In a field attraction test using traps with synthetic pheromone components, P. hybneri was attracted to (E)-2-hexenyl (E)-2-hexenoate, a component of the R. clavatus pheromone. Other components had neither an additive nor a synergistic effect on the attraction of P. hybneri. Neither (E)-2 hexenyl (E)-2-hexenoate nor other components of the R. clavatus pheromone were detected in volatiles or whole-body extracts of P. hybneri adults by gas chromatographic analysis. In addition, (E)-2-hexenyl (E)-2-hexenoate could not be found in volatiles of soybean plants. Therefore, it appears that P. hybneri responds to a component of the R. clavatus pheromone that is not emitted by P. hybneri itself. We discuss this interspecific pheromone cross-attraction of the soybean bug and hypothesize that P. hybneri utilizes the pheromone of its competitor as a kairomone for host location. PMID- 16705487 TI - Immunologic effector mechanisms of a standardized mistletoe extract on the function of human monocytes and lymphocytes in vitro, ex vivo, and in vivo. AB - Even though mistletoe extracts have been in clinical use for centuries their exact mode of action is still unknown. Currently, the application scheme for registered preparations is a dose-escalating scheme to thus reduce side effects. In this study, healthy controls and patients were evaluated for their immunologic response to treatment with a standardized mistletoe extract (Iscador). It shows a strong effect as adjuvant that induces TNF-alpha and IL-12, which was partly mediated via CD14. Desensitization of the TNF-alpha response could be shown after repeated application in vitro and in vivo. Furthermore, Iscador induces a specific lymphocyte sensitization upon multiple injections and production of IgG1 and IgG3 -mistletoe antibodies. Remarkably, a systemic bystander effect (heterologous immunity against other recall antigens) was observed after long term treatment. In conclusion, dose-escalation reduces the monocyte-related clinical side effects. A T-lymphocyte sensitization stimulates mainly a specific Th1 response. The most interesting clinical long-term effect is the bystander stimulation of various memory T cells that might mediate in vivo antitumor and antiinfectious T-cell response under mistletoe-extract immunization. PMID- 16705489 TI - Molecular modeling of the calmodulin binding region of calcineurin. AB - The Homology module within Insight-II was used to model residues 374-420, sequences missing in the coordinates of resolved structure of the catalytic subunit of calcineurin. The modeling was done in two segments. The calmodulin binding region from residues 389 to 420 was modeled based on the structure of two other proteins having calmodulin binding domains with the same 1-8-14 structural motif as calcineurin. The link region (residues 374-389) between the calmodulin binding region and the solved core sequence was generated as a random loop and two residues at the C-terminal end of the sequence were added to the model using the EndRepair function within Homology. The model was refined using the Discover module of Insight-II with energy minimization. The Builder module was used to merge the modeled regions with the solved structure of calcineurin (residues 14 373). A final refinement step was done for the joined calcineurin model. From the model, it was predicted that the calmodulin and cyclophilin binding regions seem to be proximal. Biochemical experiments provided evidence that cyclosporin-A influenced calmodulin binding and activation of calcineurin consistent with overlapping binding regions. PMID- 16705491 TI - Cognitive behavioural therapy increases re-employment of job seeking worker's compensation clients. AB - INTRODUCTION: The aim of this study was to determine whether cognitive behavioural therapy (CBT) would enhance employment outcomes in worker's compensation clients who were seeking employment. METHODS: Participants were randomly assigned to a standard job search assistance group that met for 4 hours/week for four weeks, or to a group that received standard job search assistance for 4 hours/week for the first two weeks and CBT for 4 hours/week for the next two weeks. Depression, Anxiety and Stress scores were measured prior to and on completion of the intervention, and employment outcomes were assessed at four- and ten-week follow-up. RESULTS: Affective states decreased and employment was found more rapidly after CBT than after standard job search assistance. CONCLUSION: These findings indicate that CBT has a useful role in the rehabilitation of people on worker's compensation who are seeking employment. PMID- 16705490 TI - Evaluation of work-related carpal tunnel syndrome. AB - OBJECTIVES: Carpal tunnel syndrome (CTS) is common in the industrial setting but there are still some advocates who argue that CTS is not a work related problem. There are also controversies about the proper way to establish the diagnosis and whether screening for CTS in the industrial setting is warranted. METHODS: A comprehensive literature review. RESULTS: The literature does demonstrate that the prevalence of CTS in the industrial setting is significantly higher than in the general population. Numerous epidemiologic studies have identified independent risk factors, which include repetitiveness of work, forceful exertions, mechanical stress, posture, and vibration as well as several personal co-factors. The strength of these associations are discussed. The diagnostic criteria for establishing work-related CTS are discussed as well as the effectiveness of various screening methods that are commonly used in the workplace. The sensitivity and specificity of nerve conduction studies to establish or confirm the diagnosis of CTS is presented along with normative data for the industrial worker. CONCLUSIONS: CTS has both work-related and personal risk factors. The diagnosis is best established using a combination of history, symptom distribution and confirmation using the relative latency of median sensory testing using normative data. Screening for CTS in the industrial setting has questionable benefit. PMID- 16705492 TI - Perceived effort and low back pain in non-emergency ambulance workers: implications for rehabilitation. AB - INTRODUCTION: This study aims to explore factors associated with low back pain (LBP) that required treatment from health care provider among non-emergency ambulance transfer workers. METHOD: A cross-sectional study was conducted to 38 workers of a major hospital in Hong Kong. The influences of four categories of risk factors (personal, physical, psychosocial, and exposure factors) in the prevalence of LBP were investigated by objective measurement and self-reported questionnaires. A modified Nordic musculoskeletal symptoms survey and sick leave record were used to document the prevalence of LBP. Univariate analyses followed by multiple logistic regression analyses were used to explore the risk factors associated with LBP cases. RESULTS: The results revealed that LBP was associated with age (OR=0.75, CI=0.56-1.00, P < 0.05), perceived effort (OR=7.95, CI=1.46 43.27, P < 0.05), job satisfaction (OR=4.18 CI=1.42-12.33, P < 0.01), and flexor peak torque at 120 degrees /s (OR=1.09 CI= 0.99-1.19, P=0.07). CONCLUSION: This study suggests that workers' perceived exertion has an valuable role in assessing risk at this workplace. A high perceived exertion at work can signal the need for work adjustment or modification to avoid progression of low back disorder to persistent pain or intense pain. The effects of work adjustment or modification in affected workers needs to be systematically investigated. PMID- 16705493 TI - Return to work expectation predicts work in chronic musculoskeletal and behavioral health disorders: prospective study with clinical implications. AB - STUDY DESIGN: Prospective cohort study with 18-month follow-up. OBJECTIVE: To investigate if long-term sick listed persons' own predictions of their future return to work (RTW) have an impact on their RTW when controlling for other established factors. METHOD: Postal questionnaires at baseline were sent to persons who had been on sick leave for more than 90 days, and were employed in five municipalities and four county councils in Sweden. A follow-up regarding their RTW was performed 18 months later. RESULTS: After 18 months 135 out of 508 persons (27%) had returned to work, full or part-time. In a multivariate logistic regression, the sick listed persons' own prediction of their RTW proved to be highly significant (OR=8.28, 95% CI: 3.31-20.69). Only six out of 132 persons with a negative view of their RTW did return to wok. Other predictive factors that were found for RTW were: being on sick leave for a period of less than 1 year (OR=2.09, 95% CI: 1.19-3.67), having less pain than persons in the quartile with most pain (OR=2.65, 95% CI: 1.21-5.81), perceiving that one was welcome back to work (OR=1.98, 95% CI: 1.10-3.58), and being under 55 years of age (OR=2.37, 95% CI: 1.07-5.23 for age between 45 and 54 years and the same trend for age below 45 years OR=1.85, 95% CI: 0.82-4.20). CONCLUSION: Persons with a positive prediction should get help to realise their potential for RTW. Offering traditional rehabilitation measures to a person with a negative prediction of his/her RTW, could be a waste of resources if done ahead of improving self confidence and view of what is possible. The problems in this group might decrease or be easier to handle if decisions about the future are taken within a year. PMID- 16705494 TI - Measurement qualities of a self-report and therapist-scored functional capacity instrument based on the Dictionary of Occupational Titles. AB - Studies provide convincing arguments to support the development of functional capacity instruments based on the Dictionary of Occupational Titles (DOT). The purpose of this study is to investigate the item-level measurement qualities of a newly developed DOT-based functional capacity instrument for clients undergoing rehabilitation treatment for back pain. Client and therapist ratings were collected on 124 clients from 27 rehabilitation sites using the newly developed Occupational Rehabilitation Data Base (ORDB) functional capacity instrument. Rasch analysis was used to investigate: (1) unidimensionality, (2) hierarchical item difficulty continuum, (3) rater severity, and 4) person-item match. Overall, the functional capacity scale of the ORDB showed good measurement qualities. All items, except the Handling item fit the Rasch measurement model. Because of high fit statistics and loading on factors independent from the remainder of the items, the "handling" item was removed, from further analyses. Separate client rated and therapist-rated instruments retained good item-level psychometrics. While client and therapist items showed similar item-difficulty hierarchical structures, clients had a tendency to be more severe in their rating and the correlation between client and therapist ratings was relatively low, 0.32. These findings suggest that Handling items should not be included as a DOT measure for clients with back pain. While the above psychometric study supports using client or therapist ratings as independent instruments, the lack of concordance between these ratings requires further investigation. PMID- 16705495 TI - Changing trends in US injury profiles: revisiting non-fatal occupational injury statistics. AB - The purpose of this paper is to review the current trends in non-fatal injury profiles of workers in the United States. It is generally accepted that occupational injury and illness rates are affected by many factors, such as the amount and quality of training, employee turnover rates, work experience, extent of mechanization and automation, job-related parameters, and worker gender. In the last decade, not only have the technologies used in the workplace changed significantly, there has been a greater awareness among employers and employees as to the importance of containing work injuries. Additionally, the extent of outsourcing for labor-intensive jobs has increased dramatically owing to cheaper labor costs in places such as China and Mexico. These changes have affected the manufacturing sector of US industry more than any other sector. How these changes have influenced the injury and illness profiles of the American worker is of considerable interest given the increased attention paid to work-workplace design, injury hazard control, and ergonomics in general. In this paper, we compare the injury and illness profiles of US workers separated by nearly a decade. The trends from early 1990s are compared to those from early 2000s. Data from the Bureau of Labor Statistics were used to compile the injury statistics. The results of our comparison show that while the absolute numbers of work related injuries and illnesses have declined over the last 10 years, the basic trends associated with different factors remain almost unchanged. The reasons for this decline are discussed in this paper. PMID- 16705496 TI - Development and validation of a short-form functional capacity evaluation for use in claimants with low back disorders. AB - OBJECTIVES: Functional Capacity Evaluations (FCE) are used for making return-to work decisions, yet FCE's modest predictive ability is currently outweighed by the administrative burden of testing. We attempted to develop a short-form FCE while maintaining comparable predictive ability. METHODS: Three databases previously created for evaluating FCE predictive validity were used. Subjects were compensation claimants with low back disorders. FCE measures included items in the Isernhagen Work Systems' FCE. Days until benefit suspension served as an indicator of return-to-work. Analysis included Cox regression. RESULTS: Three items, floor-to-waist lift, crouching, and standing, were maintained in the short form FCE. The short-form FCE was found to predict comparably to the entire FCE protocol in two validation cohorts (R (2) difference<3%). Subjects meeting job demands on all three items consistently experienced faster benefit suspension. CONCLUSION: A short-form FCE for determining future work status in claimants with low back disorders was developed. A substantially abbreviated FCE may offer an efficient alternative. PMID- 16705497 TI - Treatment of impingement syndrome: a systematic review of the effects on functional limitations and return to work. AB - The goal of this systematic review is to evaluate the effectiveness of different treatments for impingement syndrome and rotator cuff tear on the improvement in functional limitations and concomitant duration of sick leave. A systematic search for clinical trials or controlled studies was conducted with the following text words: should*, rotator cuff, impingement, work, sick leave, disabilit*, function*. Nineteen articles were included in this review. For functional limitations, there is strong evidence that extracorporeal shock-wave therapy is not effective, moderate evidence that exercise combined with manual therapy is more effective than exercise alone, that ultrasound is not effective, and that open and arthroscopic acromioplasty are equally effective on the long term. For all other interventions there is only limited evidence. We found many studies using range of motion and pain as outcome measures but functional limitations were less often used as an outcome measure in this type of research. Duration of sick leave was seldom included as an outcome measure. PMID- 16705498 TI - Neighborhood disadvantage, parent-child conflict, neighborhood peer relationships, and early antisocial behavior problem trajectories. AB - This study examined relations among neighborhood disadvantage, parent-child conflict, deviant peer involvement in the neighborhood, and early-starting antisocial trajectories. Antisocial group patterns were identified in 218 low income boys followed from ages 5 to 11, and neighborhood and family variables were evaluated as predictors in early and middle childhood. Four trajectory groups emerged: one increasing pattern that corresponded with developmental theories of early-starting antisocial behavior; one with initially high and decreasing problems over time; and two low antisocial groups. Parent-child conflict and neighborhood disadvantage were significantly associated with trajectory patterns, with youth in the 2 higher antisocial behavior groups characterized by more neighborhood problems and parent-child conflict than other groups. The results suggest that in early childhood, neighborhood disadvantage and family conflict place children at risk for early-starting trajectories, and that involvement with deviant peers in the neighborhood takes on an increasingly important role in patterns of antisocial behavior over middle childhood. PMID- 16705501 TI - Prevalence of visual handicap in the elderly institutionalised population in the Geneva area. AB - PURPOSE: The aim of this study was to establish the prevalence of visual impairment in the elderly population in geriatric homes in an urban area. PATIENTS AND METHODS: A sample of 96 patients from 8 geriatric homes ["Etablissements Medico-Sociaux" (EMS)] in the Geneva area were examined. The NEI VFQ-25 modified questionnaire was used. Visual acuity examination (far and near), contrast visual acuity measurement and enlargement need were administered by low vision trainers. RESULTS: Of the 96 patients 42 % were partially sighted and 3 % blind. Self assessment of visual performance disclosed that 31 % of the patients complained about bad vision but only 12 % were worried about their vision. Cataract, age-related macular degeneration, glaucoma and diabetes were the most frequent diagnosis associated to visual disability. DISCUSSION: The prevalence of the visual handicap in the elderly geriatric population is very high, but most of the patients are neither aware, nor worried about their handicap. This maybe related to associated general health disabling diseases. PMID- 16705499 TI - Relations of parenting and negative life events to cognitive diatheses for depression in children. AB - In a sample of 299 children (grades 2, 4, and 6), we examined parenting and negative life events as predictors of depressive cognitions, specifically low self-perceived competence, depressive cognitive schemas, and depressogenic attributional style. We also examined developmental trends in these relations. Children completed measures of parenting, negative life events, and depressive cognitions. Parents also completed measures of parenting and negative life events. Consistent with our hypotheses, negative parenting and negative life events corresponded with higher levels of depressive cognitions, whereas positive parenting corresponded with lower levels of depressive cognitions. The relations between negative parenting and negative automatic thoughts were stronger for older children. Theoretical and practical implications are discussed. PMID- 16705503 TI - [Chronic (kerato-) conjunctivitis refractory to therapy in children]. AB - BACKGROUND: Chronic conjunctivitis and keratoconjunctivitis account for a significant ocular morbidity. Early diagnosis and appropriate management are essential to avoid persisting structural damage and visual loss. In children, the correct diagnosis is frequently delayed because of the low individual experience with pediatric ocular inflammation, the uncommon clinical manifestations and the rarity of some conditions. This study aims to identify the problems associated with the diagnosis of chronic pediatric (kerato-)conjunctivitis. PATIENTS AND METHODS: 48 consecutive tertiary referrals (median age: 8.5 years) with chronic conjunctivitis or keratoconjunctivitis were studied. The ocular inflammation of all patients was denoted by their referring ophthalmologists as "chronic conjunctivitis refractory to therapy". The median time since disease onset was 23 months (range: 3 - 118). On average, 2.8 (range: 2 - 5) ophthalmologists were seen before the final diagnosis was made. A standardized protocol was used to classify and diagnose the ocular inflammation. Laboratory investigations were carried out to confirm the diagnosis in 20 out of 48 patients. RESULTS: In 33 out of 48 patients treatment failure was due to an inappropriate diagnosis. The most frequent diagnosis were Staphylococcus-associated inflammation (n = 21) and vernal keratoconjunctivitis (n = 12). Viral infection causing molluscum contagiosum was the most frequent condition that was missed (n = 7). Ligneous conjunctivitis (n = 2) was not recognised by the 9 ophthalmologists who were previously in charge of the treatment. Significant corneal involvement was found in 24 (50 %) patients. CONCLUSION: History taking and a thorough clinical examination of the entire ocular surface allow the correct diagnosis of and therapy for chronic surface inflammation in almost all pediatric patients. Subtle clinical changes have to be sought actively to avoid misdiagnosis. Such changes include lesions in the anterior lid margin, collarettes, follicules, papillae, and superficial punctate keratopathy. PMID- 16705502 TI - [Povidone-iodine for treatment of adenoviral conjunctivitis: an in vitro study]. AB - BACKGROUND: Adenoviral conjunctivitis causes high socioeconomic costs due to high contagiousness and therefore the need for extended quarantine. To date the only potentially active, topical antiviral agent is povidone-iodine (PVI). The aim of this study was to investigate the effect of diluted PVI on free adenovirus and adenoviral infected cells as well as to evaluate the cellular toxicity of PVI on non-infected cells. MATERIAL AND METHODS: PVI was diluted to a final concentration of 0.0008 %. Virucidal activity was measured IN VITRO using adenovirus 8 and A549 human epithelial cell cultures. Cytotoxicity effects on healthy cells after short- and long-term exposure to diluted PVI were measured in A549 cell cultures. RESULTS: Exposure to PVI at a concentration of 1:10 (0.8 %) completely extinguishes infectivity of free adenovirus after an exposure time of 10 minutes. PVI is less effective against intracellular adenovirus resulting in a decreased infectivity and viral activity for approximately one day with a narrow spectrum between toxicity and virucidal activity. Healthy epithelial cells can be exposed to PVI for up to 6 hours without a cytotoxic effect. CONCLUSIONS: PVI is highly effective against free adenovirus but less effective against intracellular adenoviral particles in already infected cell. Short- and long-term exposure of PVI causes little cytotoxicity for healthy cells. Therefore, administration of diluted PVI at a concentration of 1:10 is a potential option to reduce contagiousness in cases of adenoviral infections. PMID- 16705504 TI - [Risk of bacterial contamination after povidone-iodine disinfection for cataract surgery]. AB - BACKGROUND: Povidone-iodine alone or in combination with antibiotics is commonly used for presurgical disinfection in cataract surgery. In spite of the use of the combination Povidone-iodine and ofloxacin, the rate of ocular contamination as assessed from surgical knives was reported to be as high as 26 %. The aim of this study was to investigate the efficacy of diluted Povidone-iodine alone for surgical disinfection. PATIENTS AND METHODS: 126 consecutive patients undergoing elective cataract surgery with a conjunctival wound and a scleral tunnel received prior to surgery a disinfection with diluted Povidone-iodine eye drops (Braunol 1:10 diluted = 0.8 % Povidone-iodine, 3 times every 5 min). To assess residual bacteria on the ocular surface after disinfection, the surgical knives for the side ports and the scleral tunnel were cultured in thioglycolate broth. RESULTS: In 8 out of 126 (6 %) patients the culture from the surgical knives revealed a positive result (89 % coagulase negative Staphylococci). Four of these 8 cases occurred during a single list. All control cultures remained negative. CONCLUSION: Diluted Povidone-iodine eye drops alone are highly effective for bacterial disinfection when applied properly. The rate of contamination using 0.8 % Povidone-iodine in our series was considerably lower as compared to that of other studies. PMID- 16705505 TI - PVR recurrence and the timing of silicon oil removal. AB - BACKGROUND: Following vitrectomy for PVR-associated retinal detachment, placement of an encircling band, filling with silicone oil (SO) and successful retinal reattachment, a recurrence of PVR can develop. Retinal redetachment after SO removal is usually due to secondary or residual PVR. We wanted to ascertain whether the anatomical and functional outcomes of surgery in patients with a reattached retina and recurrent PVR can be improved by delaying the removal of SO. PATIENTS AND METHODS: 112 consecutive patients with PVR-associated retinal detachment who had undergone vitrectomy with SO filling, were monitored for at least 6 months after SO removal. Prior to SO removal, the retina posterior to the encircling band had to be completely reattached. Patients who developed PVR after SO filling were divided into two groups according to the duration of SO retention: 12 - 18 months (group 2: n = 48); > 18 months (group 3: n = 21). Individuals without PVR recurrence after SO filling and in whom the SO was consequently removed within 4 - 12 months served as control (group 1: n = 43). Anatomical success, intraocular pressure (IOP) and best-corrected visual acuity (BCVA) served as the primary clinical outcome parameters. RESULTS: Six months after SO removal, the anatomical success rates (86.3 %, 88.8 % and 84.6 %, in groups 1, 2 and 3, respectively; log rank = 0.794) and the BCVAs (p = 0.861) were comparable in the three groups. Mean IOP (p = 0.766), and the frequency of complications such as PVR recurrence (p = 0.936), bullous keratopathy (p = 0.981) and macular pucker (p = 0.943) were likewise similar. Patients in whom SO was retained for more than 18 months had the highest IOPs and required the heaviest dosage with anti-glaucoma drugs. CONCLUSIONS: In patients who develop a recurrence of PVR after vitrectomy and SO filling the surgeon can observe and treat retinal changes for up to 18 months without impairing the anatomical and functional outcomes. The retention of SO for more than 18 months does not improve the anatomical outcome. However, it can impair the functional outcome by precipitating the development of a persisting secondary glaucoma. PMID- 16705506 TI - [Interferon causes ischemic ocular diseases--case studies and review of the literature]. AB - BACKGROUND: Reports of interferon-associated retinopathy mainly originate from Japanese working groups. Only a few cases have been described in Europe. PATIENTS AND METHODS: We have performed a retrospective analysis of the patients who were examined in the University Eye Clinic in Mainz complaining of visual loss associated with systemic interferon therapy in the time span between January 2001 and December 2003. RESULTS: 12 patients (age 35 - 67 years), undergoing treatment with interferon alpha for hepatitis C, metastatic renal cell carcinoma, leukaemia and malignant melanoma of the skin, complained of blurred vision 2 - 10 weeks after the start of interferon therapy. Fundus examination revealed retinal haemorrhages and cotton-wool spots in all eyes. Additionally in some eyes macular edema, retinal arteriolar occlusion or anterior ischemic optic neuropathy was seen. As underlying risk factors we found diabetes mellitus (6 x) and systemic hypertension (3 x). After interruption (10 x) or dose reduction (2 x) of interferon therapy in all eyes the ischemic changes disappeared rapidly. Permanent visual loss or visual field defects remained in 3 out of 24 eyes. CONCLUSIONS: Patients under interferon therapy should be monitored closely by an ophthalmologist, especially those with systemic vascular risk factors. In the case of development of ischemic microangiopathy of the retina or optic nerve, interferon therapy should be stopped or dose reduction is recommended. PMID- 16705507 TI - Photodynamic therapy in the treatment of chronic central serous chorioretinopathy. AB - BACKGROUND: Central serous chorioretinopathy (CSC) is usually a benign disorder which resolves spontaneously, and requires no treatment. Nevertheless, in cases of chronic or recurrent detachment of the neurosensory retina a durable decrease of the visual acuity may be measured due to lesion of the photoreceptors. To avoid this evolution we performed a pilot study to assess the effect of photodynamic therapy (PDT) in patients with CSC without clinical normalization 6 months after the begin of the symptoms. PATIENTS AND METHODS: We report on 14 eyes in 13 patients presenting a chronic CSC without leaking point accessible for focal laser photocoagulation. The diagnosis was confirmed by fluorescein angiography (FA), and optical coherence tomography (OCT). PDT with verteporfin was performed according to the protocol used for treating choroidal neovascularization in age-related macular degeneration. RESULTS: One month after PDT, leakage on FA and detachment of the neurosensory retina on OCT had disappeared, and visual acuity had improved in all patients. CONCLUSIONS: The mechanism of action of PDT in chronic CSC is still hypothetical. PDT should decrease the passage of fluid towards the retina by affecting the choroidal blood flow, and allow a better resorption of the subretinal fluid. PDT could be an alternative to treat patients with chronic CSC. PMID- 16705508 TI - Macular atrophy after combined intravitreal triamcinolone acetonide (IVTA) and photodynamic therapy (PDT) for retinal angiomatous proliferation (RAP). AB - BACKGROUND: Retinal angiomatous proliferation (RAP) is a variant of exudative AMD that appears to respond poorly to standard PDT. This pilot study explores the potential efficacy and safety of combined intravitreal triamcinolone acetonide (IVTA). PATIENTS AND METHODS: In a prospective interventional case series 10 eyes of 8 patients with RAP were treated with combined IVTA/PDT. RESULTS: After 3 months there was no evidence of significant exudative activity in 7/10 eyes treated with IVTA/PDT, three eyes required re-treatment. After 6 months 4/10 eyes were "dry" on fluorescein angiography, visual acuity was stable (loss of less than 3 Snellen lines) or had improved in 5/10 eyes. 5/10 eyes had lost 3 or more lines, due to central macular atrophy in 4/5 eyes and secondary to a progressive exudative lesion in 1/5 eyes. CONCLUSIONS: Intravitreal steroids combined with PDT appear to be a step ahead in the treatment of RAP. Our results, however, raise the question whether combined treatment may create more atrophy leading to vision loss in some cases. PMID- 16705509 TI - [Corticosteroids in giant cell arteritis: primum nil nocere?]. AB - BACKGROUND: Steroids are the treatment of choice for giant cell arteritis but bear the risk of serious side effects. PATIENTS AND METHODS: We carried out a retrospective study on 34 patients with documented giant cell arteritis (24 with ocular involvement) by means of a questionnaire sent to the treating physicians. RESULTS: After a mean follow-up of 48 months, side effects occurred in 90 % of the patients. The most frequent were weight gain (> 50 %) and osteoporosis (> 40 %, F >> M). Side effects were more common in patients with ocular involvement and in women. Severe complications were significantly more frequent in patients with ocular involvement. CONCLUSIONS: Side effects are the rule and not the exception in the treatment of giant cell arteritis. They can affect quality of life. Physicians should bear them in mind as many are preventable and/or treatable. PMID- 16705510 TI - [Binocular diplopia after cataract operation]. AB - BACKGROUND: The development of binocular diplopia after cataract surgery is a serious complication. The present retrospective study aims to improve our understanding of this phenomenon and its prevention. PATIENTS AND METHODS: A series of 11 patients with binocular diplopia after cataract surgery under local retrobulbar anaesthesia is presented. The patients were followed over a period ranging from 1 to 31 months. RESULTS: Three types of disorders were found: muscular or nervous injuries (7/11), decompensated latent heterophorias (2/11), and disclosed previous strabismus with abnormal retinal correspondence (2/11). The treatment was surgical for 3 patients, and a prismatic correction was prescribed for 6 patients. Spontaneous recovery was seen in 5 patients. CONCLUSION: Fortunately, binocular diplopia after cataract surgery is a rare complication, with multiple aetiopathologies. Muscular injuries, related to the anaesthesic technique, are the most frequent disorders responsible for diplopia and often require strabismus surgery. PMID- 16705511 TI - [Surgical correction of subjective excyclotorsion by tucking the anterior part of the superior oblique muscle tendon]. AB - BACKGROUND: Asymmetrical folding of the anterior portion of the superior oblique muscle is a simplified variant of the Harada-Ito procedure that is supposed to selectively correct ocular torsion. PATIENTS AND METHODS: This surgical technique was applied to 22 patients suffering from bilateral acquired IV nerve palsy. Ocular torsion was evaluated with the Harms tangent screen. Pre- and post operative torsional values were compared. Post-operative follow-up was 12 months. RESULTS: Pre-operatively, mean excyclotropia was 8.8 degrees in primary position and 15.9 degrees in downgaze. Post-operatively, these values decreased to 1.5 degrees and 4.1 degrees, respectively. An enlarged field of binocular single vision, extending beyond 10 degrees inferiorly, was obtained in 17/22 patients. CONCLUSIONS: The selective technique of superior oblique muscle anterior tendon tucking allows one to efficiently correct the ocular torsion induced by acquired bilateral IV nerve palsy. PMID- 16705512 TI - [Corneal and conjunctival findings in systemic silver intoxication]. AB - BACKGROUND: Chronic silver intoxication is a rare disease and therefore the typical ocular findings may be missed. Based on a case with severe intoxication, the clinical and histological findings as well as the prognosis in argyria are presented. HISTORY: A 33-years-old-employee of a battery production plant developed a biopsy proven systemic argyria. On slit lamp examination the conjunctiva showed a dark, blue-grey discoloration in the areas of the tear flow and the small conjunctival arteries. Diffuse silver deposits were noticed on the level of Descemet's membrane without endothelial damage. Silver deposits were also visible in the trabecular meshwork. Chemical reaction of the silver particles upon exposure to UV light results in irreversible tissue discoloration. Without options for an effective treatment, early diagnosis and prevention of overexposure are most important. CONCLUSIONS: The typical ocular findings in systemic argyria are helpful in occupational medicine for establishing the correct diagnosis. PMID- 16705513 TI - [Leber's hereditary optic neuropathy in malnutrition: a case report]. AB - BACKGROUND: We present a case of Leber's hereditary optic neuropathy (LHON) manifested by malnutrition, tobacco and alcohol abuse. HISTORY AND SIGNS: We report on a 36-year-old patient with alcohol and tobacco abuse for years. 5 months before manifestation of LHON, the alcohol abuse was stopped. Because of congenital cataract the reduction in visual acuity, which occurred after alcohol consumption was stopped, was first misinterpreted. Findings at first exam: visual acuity right eye 0.2, left eye 0.05, temporal optic disc pallor left > right, vitamin B12 and folic acid at the lower level. Molecular genetic analysis: LHON mtDNA 11 778 mutation. Therefore, a pure tobacco-alcohol optic neuropathy could be excluded. THERAPY AND OUTCOME: Under high-dose vitamin-B complex substitution the visual acuity did not improve in the next 6 months. CONCLUSIONS: The cause of the manifestation of the LHON is unknown and multifactorial. In this case the manifestation was caused by malnutrition and tobacco abuse. PMID- 16705514 TI - Evanescent white linear flecks and posterior microphthalmos: new features of a recently established disease. AB - BACKGROUND: Posterior microphthalmos is a rare autosomal recessive condition with normal anterior segment and small posterior segment resulting in axial hyperopia and retinal folding. HISTORY AND SIGNS: The proband from a consanguineous Turkish family was clinically investigated at 5 months of age and followed for the next 8 years. At five months of age, refraction revealed a severe hyperopia of + 10 diopters in both eyes (OU) with a mean axial length of 16.5 mm. Fundus examination showed a dystrophic maculo-papillar bundle. Multiple whitish deep intraretinal linear lesions sparing the macula were present in the mid-periphery with no preferential orientation. THERAPY AND OUTCOME: By 8 years of age, visual acuity was 20/25 in the right eye (OD) and 20/30 in the left eye (OS) with S + 14 OU. The whitish linear lesions had disappeared and were replaced by retinal pigment epithelium alterations. Optical coherence tomography (OCT) revealed a thickened retina with normal appearance at the site of the linear lesions. CONCLUSIONS: This is the first report of long-term follow-up and OCT findings in this rare ocular developmental abnormality. PMID- 16705515 TI - Rehabilitation of orbital cavity after total orbital exenteration using oculofacial prostheses anchored by osseointegrated dental implants posed as a one step surgical procedure. AB - BACKGROUND: Total orbital exenteration is a radical surgical procedure, which typically involves the removal of the entire contents of the orbit including the periorbita, leaving the patient with a deep orbital deformity and results in devastating cosmetic, functional, and psychological consequences requiring difficult and challenging procedures for oculoplastic surgeons. Oculofacial prostheses retained by endosseous dental implants present an attractive and viable alternative when aesthetic and functional demands are beyond the capacity of local reconstructive efforts. PATIENTS AND METHODS: A 72-year-old woman presenting a malignant melanoma of the right eyelids and a 77-year-old man presenting a sebaceous carcinoma of the left upper eyelid underwent a total exenteration followed by positioning of endosseous implants (Straumann system Dental implants) as an integrated one-step combined surgical procedure. The oculofacial prosthesis was placed after epithelialization of the orbital cavity. RESULTS: The implants were perfectly osseointegrated without any complications, providing sufficient retention of the prostheses. A satisfactory aesthetic outcome has been achieved for both patients. CONCLUSIONS: Oculofacial prostheses anchored by osseointegrated dental implants placed as one-step surgical procedure ensure an adequate aesthetic result as well as a considerably decreased rehabilitation time and present a satisfactory solution when reconstruction is not a suitable option. PMID- 16705517 TI - Acquired unilateral pseudoptosis: a presentation of silent sinus syndrome. AB - BACKGROUND: Acquired unilateral ptosis usually results from either levator palpebrae disinsertion, eyelid disorder, myasthenia gravis, mitochondrial myopathy, Horner's syndrome, or third nerve palsy. In rare cases, an orbital process is responsible for ptosis or pseudoptosis. HISTORY AND SIGNS: A healthy 31-year-old patient complained for 6 months of right eyelid ptosis. On examination, there was 1 mm right upper eyelid ptosis and 1 mm of right enophthalmos. Upon downgaze, an apparent right eyelid retraction was present. Magnetic resonance imaging showed filling and retraction of the right maxillary sinus with inferior displacement of the right orbital floor. THERAPY AND OUTCOME: Surgical management was performed with a favourable outcome. CONCLUSIONS: Silent sinus syndrome is an asymptomatic chronic maxillary sinus atelectasis, resulting in ipsilateral enophthalmos, hypoglobus, and occasionally eyelid pseudoptosis. Silent sinus syndrome should be incorporated in the differential diagnosis of acquired unilateral ptosis. PMID- 16705516 TI - [Orbital MALT lymphomas: clinicopathological correlation]. AB - BACKGROUND: Extranodal MALT lymphomas are slow growing tumors of B-cell origin which may be found in the orbit. They are associated with mucosa and epithelial structures. PATIENTS AND METHODS: We present eight patients with biopsy confirmed orbital MALT Lymphomas. The diagnostic imaging techniques are described. Histopathological and immunohistological analysis showed typical findings of MALT lymphomas. RESULTS: After staging six patients had radiation therapy. Two patients refused treatment due to lack of discomfort. CONCLUSIONS: MALT lymphomas should be considered in the differential diagnosis of orbital tumors. While ultrasonography and MRI are needed to determine the extension of these tumors, their identification requires excision or biopsy with histological/immunohistochemical analysis, especially in view of new treatment options. PMID- 16705518 TI - Unspecific redness of the eyelid--two malignant tumours. AB - BACKGROUND: Chronic, non- or little pigmented reddish or brownish lesions on the lid margin and the surrounding skin are frequently underestimated and thus carry a clear risk of malignancy. HISTORY AND SIGNS: A 61-year-old lady was referred with a chronic, reddish lesion in the medial third of the left lower lid after a topical therapy over a period of 4 months had not improved the situation. She noticed the lesion for the first time 10 previously. Recurrent trichiasis and a slight ulceration had developed during the last year. THERAPY AND OUTCOME: After an unclear result of the biopsy (2)/ (3) of the left lower lid were removed because of atypical cells ascending into the epidermis. A superficial spreading melanoma 0.4 mm depth in the Breslow classification was detected and right next to it a superficial basal cell carcinoma. The lid defect was reconstructed with a tarsoconjunctival flap and a free full thickness skin graft as described by Hughes. CONCLUSION: Chronic redness at or around the lid margin which does not respond to therapy should be biopsied as malignant tumours may well be hidden behind it. PMID- 16705519 TI - Spontaneous subconjunctival orbital fat prolapse: presentation of four cases. AB - BACKGROUND: Subconjunctival orbital fat prolapse is a benign entity characterized by orbital fat herniation through a dehiscence in Tenon's capsule, usually between the superior and lateral recti muscles. It is often associated with ocular trauma or surgery, although spontaneous cases have also been reported with a predilection for obese individuals. PATIENTS AND METHODS: A retrospective study of four subconjunctival orbital-fat prolapse cases with no history of ocular surgery or trauma was performed. A trans-conjunctival surgical approach with Tenon's capsule repair was offered to all subjects. Differential diagnosis is discussed. RESULTS: The diagnosis of subconjunctival orbital fat prolapse was confirmed in all cases by histopathological examination. Functional and aesthetic postoperative results were excellent, with no recurrences during follow-up, for all cases. CONCLUSIONS: Although rare, subconjunctival orbital fat prolapse should be recognized. Surgical management offers excellent results. PMID- 16705520 TI - [Long-term follow-up and angiographic findings in serpiginous choroiditis]. AB - BACKGROUND: Long-term follow-up and angiographic findings of serpiginous choroiditis, a chronic disease with recurrences of unknown origin, are reported. PATIENTS AND METHODS: A retrospective analysis of the data of 4 patients with serpiginous choroidopathy, who were treated in the eye clinic of Mainz in the past 10-18 years was made. Progression of disease was documented by photography, as well as fluorescein and indocyanine green angiography. Patients with acute lesions were treated with steroids per os for some weeks. RESULTS: All 8 eyes developed 2-7 recurrences with progression of the chorioretinal scars. In cases of macular involvement the functional results were worse. No choroidal neovascularizations were seen. During follow-up 3/8 eyes became blind, 4/8 eyes retained reading vision. Indocyanine green angiography usually demonstrated many more lesions than those seen with fluorescein. In 3 cases acute choroidal lesions were detected only with indocyanine green angiography whereas no sign of inflammation was demonstrated by fluorescein angiography. CONCLUSION: The functional prognosis of serpiginous choroiditis is bad because all eyes developed at least 2 recurrences during follow-up. Use of indocyanine green angiography may help to better describe the full extent of the disease, even before it is seen with fluorescein angiography. It can help the observer to determine the precise clinical progression of the disease. PMID- 16705521 TI - [Treatment of uveitis-associated refractory ocular hypotony with topical ibopamine]. AB - BACKGROUND: Ibopamine is a non-selective dopamine- and adrenalin-receptor agonist that has been shown to cause pupillary dilation and an increase in aqueous humour secretion. This novel drug can be used as a mydriatic agent, as a provocative test in open-angle glaucoma, and for the treatment of persisting ocular hypotony. HISTORY AND SIGNS: This 47-year-old man had a history of uveitis associated with Crohn's disease. Six years after deep sclerectomy for uveitic secondary glaucoma, he developed severe hypotony in his left eye with drop of visual acuity (VA). The hypotony did not respond to topical steroid treatment. 2 % Ibopamine solution was ordered t. i. d. concomitant to 1 % prednisolone acetate. THERAPY AND OUTCOME: Intraocular pressure (IOP) began to rise after 3 weeks of Ibopamine treatment and returned to normal (12 mmHg) with continuous recovery of VA after 8 weeks. Ibopamine was discontinued at an IOP of 16 mmHg after a course of 12 weeks. IOP and VA remained stable during the 12-month follow-up period. CONCLUSIONS: Ibopamine 2 % eye drops in combination with topical steroids are a therapeutic option in uveitis-associated ocular hypotony. PMID- 16705522 TI - Blood-brain barrier disruption associated with topiramate-induced angle-closure glaucoma of acute onset. AB - BACKGROUND: Topiramate (Topamax(R)) is an anti-epileptic drug of the sulfamate group used secondarily for bipolar disease. HISTORY AND SIGNS: One week after initiation of topiramate treatment for a bipolar disorder, a 57-year-old man presented with blurred vision. Clinical examination revealed a bilateral conjunctivitis, areflexic mydriasis, severe anterior chamber shallowing, with a myopic shift and vitritis. THERAPY AND OUTCOME: A spinal tap revealed an increased protein content of 1581 mg/L on cerebrospinal fluid (CSF) analysis, being compatible with a rupture of the blood-brain barrier (BBB). UBM exposed bilateral ciliochoroidal effusions with secondary angle-closure. Topiramate was promptly discontinued, whereas visual acuity, intraocular pressure (IOP), and anterior and posterior segments anatomy normalized within 1 week. One month later, bilateral iris atrophy was present. CONCLUSION: The presence of BBB disruption with increased protein content in CSF with simultaneous blood ocular barrier breakdown may suggest a common inflammatory mechanism. PMID- 16705523 TI - Autoimmune retinal dysfunction in sarcoid chorioretinopathy. AB - BACKGROUND: Visual loss, photophobia and night-blindness in a middle-aged patient with a previous history of cancer are highly suggestive of cancer-associated retinopathy. HISTORY AND SIGNS: A 63-year-old man complained of slowly progressive bilateral visual loss, photophobia and night-blindness. His past medical history was remarkable for surgical and medical treatment of a carcinoma of the base of his tongue nine years earlier. Visual acuity was 0.3 RE and 0.15 LE, with constricted visual field in both eyes. Ocular examination and fluorescein angiography were non-relevant. Electroretinography revealed diffuse cone and rod dysfunction. ICG angiography showed multiple small widespread hyperfluorescent spots. A chest CT scan revealed multiple focal lesions which were biopsied. Histological diagnosis was sarcoidosis. THERAPY AND OUTCOME: Upon systemic steroid therapy, visual function gradually improved, and ICG anomalies disappeared. Vision recovered to 1.0 RE and 0.8 LE with normal visual field in both eyes. CONCLUSION: Sarcoidosis may present as an autoimmune retinal dysfunction with photophobia and night blindness as the presenting symptoms. PMID- 16705524 TI - [Utility of cerclage surgery of rhegmatogenous retinal detachments: a retrospective study of 41 cases]]. AB - BACKGROUND: 360 degrees encirclement for rhegmatogenous retinal detachment (RD) was introduced over 50 years ago as a method for closing retinal breaks and relieving vitreous traction. In the absence of acknowledged guidelines, current usage varies widely. PATIENTS AND METHODS: All 360 degrees encircling bands for RDs performed between 1999 and 2004 were reviewed. Encircling procedures for phakic and recurrent RDs were analysed. RESULTS: 41 encircling bands for RD were identified from the records of 875 RD cases. Median follow-up was 6 months (range: 1 - 36). Male patients accounted for 58.5 % of the procedures. Mean age was 51.51 +/- 15.97 years. 37/41 phakic and 18/41 recurrent RDs were encircled. RD recurrence in encircled eyes occurred in 42 % of phakic eyes and in 33 % of eyes re-operated for recurrent RD. At the time of recurrence, proliferative vitreoretinopathy (PVR) was present in 41.6 % of phakic eyes, and 66 % of recurrent RD eyes. Preoperative visual acuity (VA) was 0.26 and post-operative VA 0.34. Post-operative complications due to the encirclement included pain in 4 eyes, scleral abscess in 2 eyes and scleral necrosis in 1 eye. CONCLUSION: This small retrospective review highlights the multiple indications for encircling bands including PVR, inferior breaks, multiple breaks and myopia. High overall recurrence rate (34 %) reflects the selection bias of this procedure for severe disease in our series. There is currently no consensus on the use of this technique in the management of RD. PMID- 16705525 TI - [Pseudophakic inferior retinal detachment or detachment with inferior breaks: clinical outcome and recurrences after primary vitrectomy-gas]. AB - BACKGROUND: This study aims to evaluate the outcome of primary vitrectomy/gas for pseudophakic retinal detachment (RD) located inferiorly or associated to inferior breaks. PATIENTS AND METHODS: We performed a retrospective, non-randomised study comprising 18 pseudophakic RD eyes with inferior RD (group A) and 19 pseudophakic RD eyes with inferior breaks (group B) treated with primary vitrectomy and gas tamponade (SF (6) 20 %). Pre- and postoperative characteristics were analysed, focusing on RD recurrences. RESULTS: In group A, RD recurred in 2/18 eyes (11 %) and was associated to proliferative vitreoretinopathy (PVR) both at first intervention and on recurrence. In group B, RD recurred in 4/19 eyes (21 %). One eye presented PVR at first intervention and all at re-intervention. Anatomic reapplication was achieved after second vitrectomy/gas in 2/4 eyes of group B, whereas long-term silicon oil tamponade was needed in 2 eyes of each group. CONCLUSIONS: This series shows a high success rate of primary vitrectomy/gas in pseudophakic RD patients with inferior RD or detachment associated to inferior breaks. Recurrences after vitrectomy are most often related to the presence of PVR. PMID- 16705526 TI - Vitreous haemorrhage in children. AB - BACKGROUND: Vitreous haemorrhage in early childhood is rare and may cause few symptoms. HISTORY AND SIGNS: Three children (aged 1, 2 and 6 years) with no history of trauma, presented with squint or unreactive pupil. Unilateral dense vitreous haemorrhage was found in all three children. THERAPY AND OUTCOME: Rapid clearing of the vitreous opacities within a few weeks made surgical intervention unnecessary in two children. A 6-year-old boy underwent a vitrectomy. Thorough investigations revealed angle recession, retinal pigment epithelium (RPE) scars and an orbital floor bone fracture, respectively, suggesting ocular trauma as the cause for the haemorrhage in all three cases. With early amblyopia treatment, the visual outcome was good. CONCLUSION: Trauma appears to be a likely cause for vitreous haemorrhage in small children. Even if there is a risk for the development of amblyopia, observation may be a reasonable approach in some cases, since even dense vitreous haemorrhage may resorb rapidly in young children. PMID- 16705527 TI - Night blindness, yellow vision, and yellow skin: symptoms and signs of malabsorption. AB - BACKGROUND: Rapidly progressing bilateral night blindness in an elderly patient suggests primarily a diagnosis of paraneoplastic retinopathy. Occasionally diffuse rod dysfunction can result from vitamin A deficiency. HISTORY AND SIGNS: A 70-year-old man complained of progressive night blindness and xanthopsia for the past 6 months. Visual acuity was 0.8 in both eyes with severe dyschromatopsia. Slit-lamp and fundus examination were normal. Visual field disclosed bilateral depression. Scotopic full-field ERG was severely reduced. The patient's medical history revealed an acute pancreatitis one year ago, followed by chronic jaundice and an increased blood bilirubin. Serum vitamin A level was decreased to 0.1 micromol/L (normal range 1.5 to 4.0). THERAPY AND OUTCOME: Intramuscular injections of vitamin A were provided. Subjective visual improvement was reported already one day after initiation of therapy. Scotopic full-field ERG was markedly improved 3 days after the injection and was only slightly subnormal 3 months later. CONCLUSIONS: In developed countries, vitamin A deficiency usually results from malabsorption syndromes and manifests initially by rod more than cone dysfunction. This diagnosis should be entertained early as vitamin A supplementation induces a rapid restoration of vision. PMID- 16705528 TI - Bilateral giant retinal tear and retinal detachment in a young emmetropic man after Jarish-Herxheimer reaction in ocular syphilis. AB - BACKGROUND: Giant retinal tears were previously described in patients affected by panuveitis. We report the case of a patient presenting giant retinal tears in both eyes affected by ocular syphilis. PATIENT AND METHOD: A 45-year-old patient presented a 5 days history of sudden bilateral drop of vision, two weeks after penicillin therapy for secondary syphilis. The best visual acuity was 0.5 RE and light perception LE. Biomicroscopy showed an intense vitritis associated with bilateral giant tear and retinal detachment. Both TPHA and VDRL were positive. Lumbar puncture showed lymphoplasmocytosis with intrathecal synthesis of IgM. RESULTS: Topical steroids treatment was applied and intravenous penicillin was given during 14 days. Pars plana vitrectomy with silicon oil tamponade was performed in association with endophotocoagulation in the left eye and cryocoagulation in the right eye. Vitreous PCR was negative. Evolution was successful with an attached retina. CONCLUSIONS: This represents the first reported case of giant retinal tear with retinal detachment in a patient presenting a syphilitic panuveitis. PMID- 16705529 TI - Re-emergence of hidden residual intraocular silicone oil bubble after previous silicone oil removal. AB - BACKGROUND: We describe the re-emergence of a hidden residual intraocular silicone oil bubble after previous silicone oil removal. HISTORY AND SIGNS: A 57 year-old patient underwent vitrectomy with silicone oil for a complicated retinal detachment. Six weeks later silicone oil was removed via the pars plana and phakoemulsification with lens implantation was performed. Subsequent clinical follow-up showed an attached retina without residual silicone oil bubbles. However, more than 2 years later the patient suffered sudden visual loss. Clinical examination showed a bubble of silicone oil wedged between a remaining layer of anterior vitreous and the capsular bag, reducing central visual acuity. THERAPY AND OUTCOME: Complete extraction of the silicone oil bubble was performed via the pars plana. CONCLUSIONS: Residual silicone oil bubbles may wedge themselves into vitreous pockets in the ophthalmoscopically invisible regions of the pars plana. Over time these bubbles may migrate into the retrolental space with obstruction of the visual axis. PMID- 16705530 TI - [Acquired temporary night blindness in vitamin A and zinc deficiency in anorexia nervosa nine years after kidney transplantation]. AB - BACKGROUND: Vitamin and trace element deficiencies may cause visual disturbances. Before starting a substitutional therapy, specific investigations should be performed. HISTORY AND SIGNS: We present a kidney-transplanted patient who suffered for 6 months from night blindness. The electroretinography showed a severe decrease of the rod activity. Except for an irregular pigmentation in the far periphery, the fundus appeared normal. These findings were indicative of a vitamin A and zinc deficiency. The laboratory tests showed decreased vitamin A and zinc serum levels. THERAPY AND OUTCOME: After three months of substitutional therapy the electroretinogram had normalized and the patient was free of symptoms. CONCLUSIONS: Visual disturbances due to vitamin and trace element deficiencies are rare in Europe. If suspected, since the diagnosis and the treatment of nutritional deficiencies are simple, specific investigations and a substitutional therapy should be initiated. PMID- 16705531 TI - Presumed macular toxicity of intravitreal antibiotics. AB - BACKGROUND: Intravitreal injection is the most effective route for administration of antibiotics in intraocular infections. PATIENTS AND METHODS: Two patients presented with a metal foreign body in the central vitreous without damage to the retina. RESULTS: Foreign bodies were removed with an intraocular magnet and limited vitrectomy was performed. Because of leukocytic infiltration of the vitreous, 1 mg vancomycin and 0.4 mg amikacin were injected in the vitreous at the end of the surgery. Postoperative visual acuity did not reach preoperative values in both patients. ERG examination revealed reduced photoreceptor activity. We speculate that amikacin may have reached the macula in a high and toxic concentration in an incompletely vitrectomized eye. CONCLUSION: The currently accepted intravitreal use of amikacin may cause retinal toxicity. PMID- 16705532 TI - Isolated IVth (trochlear) nerve palsy due to basilar artery dolichoectasia. AB - BACKGROUND: Dolichoectasia (elongation, dilatation and tortuosity) of the basilar artery can cause an isolated cranial neuropathy. The trigeminal nerve and facial nerve are most frequently affected. Dysfunction of one of the ocular motor cranial nerves due to basilar artery dolichoectasia is uncommon, and an isolated IVth (trochlear) nerve palsy has not been previously described in the literature. HISTORY AND SIGNS: Two men, ages 70 and 59 years, respectively, presented with vertical diplopia due to a IVth nerve palsy. In one patient, the onset of the IVth nerve palsy was painless and gradual and in the other patient, the onset was acute and associated with periorbital pain. Neuroimaging in both patients revealed pathological tortuosity of the basilar artery around the midbrain and displacement of the artery toward the side of the affected trochlear nerve. THERAPY AND OUTCOME: The patients were observed clinically. One patient had gradual worsening of his palsy for three and one-half years then suffered a stroke. The second patient whose IVth nerve palsy had an acute onset experienced spontaneous resolution of his palsy but later developed dysfunction of other cranial nerves. CONCLUSIONS: Basilar artery dolichoectasia should be considered in the differential diagnosis of an isolated IVth nerve palsy. The clinical course may be variable, and the prognosis is not always benign. PMID- 16705533 TI - [Visual improvement of an amblyopic eye in an adult patient after visual loss in the non-amblyopic eye]. AB - BACKGROUND: The sensitive period for a successful amblyopia treatment is limited to the age of 11 to 13 years. HISTORY AND SIGNS: We present a 60-year-old patient with complete visual loss on his dominant eye after retinal arterial occlusion. The fellow eye had a known severe amblyopia with a corrected visual acuity of 0.1. THERAPY AND OUTCOME: After retinoscopy the patient received a full correction for his amblyopic eye and the vision increased to 0.25. After three months follow-up visual acuity was 0.5 for single optotypes. CONCLUSIONS: Even in adulthood a attempt at full correction of an amblyopic eye after loss of vision in the dominant eye should be performed for optimising the quality of life. PMID- 16705534 TI - [Aetiopathogenesis of diabetes type 1 and 2: what can we learn from epidemiological data?]. PMID- 16705536 TI - [Severe diabetic fetopathy due to undiagnosed gestational diabetes mellitus]. AB - HISTORY AND FINDINGS: A 39 year-old pregnant woman (with one healthy 11 year-old daughter) presented at the antenatal care clinic for routine non-stress test (NST) at the due date. The NST demonstrated normal fetal heart pattern. No anamnestic or clinical risk factors for diabetes during the course of pregnancy were noted in the case file. Ultrasound examinations up to 30 weeks of gestation had shown fetal growth appropriate for gestational age. Two days later, the NST was non-reacting with almost no heart rate variability, indicating an acute fetal risk. While the condition of the fetus was being further assessed by Doppler sonography, a fall in fetal heart rate prompted an emergency section for fetal bradycardia. The male newborn had marked macrosomia (birth weight 5180 g). The pH in umbilical artery blood of 6,77 indicated severe intrauterine hypoxia. Apgar score was 0/1/5. Absence of heart beat and of spontaneous breathing required resuscitation. Echocardiography revealed a hypertrophic cardiomyopathy with dysfunction of the right ventricle and hepatomegaly indicating diabetic fetal pathology. The placenta displayed typical signs of carbohydrate disorder. The maternal 75 g oral glucose tolerance test confirmed the suspicion of maternal diabetes (glucose values 96/210/215 mg/dl). TREATMENT AND COURSE: During the next the day the infant developed multi-organ failure involving the liver and kidneys, myocardial infarction with pericardial effusion and myoclonic seizures. His condition improved slowly under measures to stabilize the circulation, substitution of clotting factors and anticonvulsive therapy. Mechanical ventilation had to be continued for 5 weeks. Examination at 4 months of age revealed marked neurological abnormalities. The mother was referred to a diabetes specialist for further management. CONCLUSION: Undiagnosed and therefore untreated severe gestational diabetes may have fatal consequences for the fetus. Expert committees of obstetricians and diabetes specialists have recommended blood glucose screening between 24 - 28 weeks of gestation of every pregnant woman as part of the routine prenatal care. PMID- 16705535 TI - [Health-related quality of life of siblings of children with type 1 diabetes mellitus]. AB - BACKGROUND INFORMATION: A chronic illness in childhood or adolescents constitutes a drastic event for the whole family, especially for the healthy siblings. This could impair their quality of life. PATIENTS AND METHODS: We designed a case control-study in three centres (Leipzig, Giessen and Augsburg) with questionaires sent to 72 diabetic children, 71 of their siblings and 63 children of neighbourhood (controls) aged between 8 and 16 years. KINDL-Questionnaire was used for the study of the health-related quality of life of children and adolescents, supplemented by other appropriate items. RESULTS: The results indicate a positive health related quality of life of the healthy siblings of families with a child who has type 1 diabetes. Differences in the quality of life depend on factors such as age and gender of the children and relate to specific items of sibling relationship (e. g. "worried about sibling", "looked after sibling"). Generally, there was a great similarity with regard to the quality of life of healthy brothers and sisters from affected families compared with children of families without a child suffering from chronic illness. CONCLUSIONS: The results indicate that the health related quality of life of healthy siblings of children with diabetes only differs insignificantly from brothers and sisters of "healthy" families. PMID- 16705538 TI - [Diabetes in old age -- do we overstrain our patients?]. PMID- 16705539 TI - [Metabolic syndrome -- myths, mechanisms, management]. PMID- 16705540 TI - [Diabetic retinopathy and maculopathy]. PMID- 16705541 TI - [Prevention studies in childhood: are they reasonable? -- Pro]. PMID- 16705542 TI - [Prevention studies in childhood: are they reasonable? -- Contra]. PMID- 16705543 TI - [Compensation claims in defiance of invalid insurance claims?]. PMID- 16705544 TI - [Quality of health care and degree of complications in a publicly known study of type 2 diabetic patients ]. PMID- 16705546 TI - The Leu7Pro polymorphism of neuropeptide Y is associated with younger age of onset of type 2 diabetes mellitus and increased risk for nephropathy in subjects with diabetic retinopathy. AB - Several studies have shown genetic predisposition for diabetic complications. The leucine7 to proline7 (Leu7Pro) polymorphism of preproNPY has been shown to be a risk factor for diabetic retinopathy in type 1 diabetes. In the current study we examined the contribution of this polymorphism on the progression of retinopathy in Caucasian type 1 and type 2 diabetes patients. Patients with type 2 diabetes and the Leu7Pro polymorphism developed retinopathy at younger age because of markedly earlier disease onset of diabetes (RC- 6.8, 95% CI-12.2 - [- 1.5]), but no association of the Leu7Pro polymorphism with the current severity of retinopathy was detected. A strong association of the polymorphism with proteinuria in type 2 diabetes patients with retinopathy could be detected (OR 3.1, 95% CI 1.1-8.8); 31% of subjects having both retinopathy and proteinuria had the polymorphism compared to only 13% of retinopathy patents without concomitant proteinuria (p = 0.032). Plasma concentrations of NPY were increased in subjects with proteinuria (79.2+/-28.4 and 64.7+/-26.2 pmol/l, p = 0.001). These results suggest that the Leu7Pro polymorphism could be used to predict earlier onset of type 2 diabetes and retinopathy, and increased risk for diabetic nephropathy. PMID- 16705548 TI - Novel mitochondrial DNA length variants and genetic instability in a family with diabetes and deafness. AB - We have identified two locations with novel multiplasmic length variants in the mitochondrial DNA of a family with diabetes and deafness. At nt568 in the D-loop, the 6-bp polycytidine tract was found to be variable in length up to a total of 12 residues. A second region with length variants was found at nt8281 in the intergenic COII-tRNA(Lys) region, which consists of two copies of the 9-bp repeat CCCCCTCTA. Only the second repeat occurs in a heteroplasmic C(9-14)A form with both T residues largely deleted. In addition, the mtDNA contained a number of new homoplasmic point mutations. Both length variants are stably inherited in a maternal way with no major changes in their length distribution. In contrast, during culture of fibroblasts from the proband the average length of the polycytidine tracts is increased at both locations indicating a fibroblast specific genetic instability. Cybrid cells containing mtDNA from the proband proliferate less efficient than cybrids with wild-type mtDNA in co-culture experiments, suggesting functional consequences of the mtDNA length variants or the additional homoplasmic point mutations. Since oxygen consumption was not severely affected, these mutation seem less detrimental for mitochondrial function than the A3243G diabetogenic mutation and most other pathogenic mtDNA mutations. The contribution of mtDNA length variants to the phenotype of members of this family is discussed. PMID- 16705547 TI - Postprandial mononuclear NF-kappaB activation is independent of the AGE-content of a single meal. AB - OBJECTIVE: Dietary uptake of Advanced Glycation Endproducts (AGE) is supposed to potentially contribute to inflammatory reactions linked to vascular dysfunction and late diabetic complications. One mechanism by which dietary AGE might exert these effects is by activation of the proinflammatory transcription factor NF kappa-B. The aim of this study was to analyze the postprandial effects of a casein meal with low or high AGE content on postprandial NF-kappaB activation in peripheral blood mononuclear cells (pBMC) of healthy volunteers. RESEARCH DESIGN AND METHODS: Casein was heated for 40 h at 50 degrees C in the presence of sorbitol or glucose, resulting in either minimal (Sorbitol [S]-casein) or large (glucose [G]-casein) amounts of AGE-modified casein. Nine healthy volunteers ate 250 g of both types of casein, whereas both meals were separated at least by 2 weeks. Plasma and pBMC were taken before and 2 h after each meal. Thereafter, the defined AGE carboxymethyllysine (CML) was determined by ELISA and Western blot. NF-kappaB activation in pBMC was assayed using Electrophoretic Mobility Shift Assays (EMSA) and Western blot analysis. RESULTS: S-casein contained only minor amounts of CML and no pentosidine, while G-casein contained large amounts of both. 2 h after ingestion, the S-casein or the G-casein-meal, both, resulted in a non-significant increase in plasma CML and in the intracellular CML-content of pBMC. This was paralleled by a highly significant increase in postprandial mononuclear NF-kappaB-binding activity. Remarkably, neither the extent of NF kappaB induction (178% for S-casein, 188% for G-casein), nor composition of the NF-kappaB heterodimer (mainly consisting of NF-kappaB p50/p65) were significantly different after intake of S-casein or G-casein. Consistently, Western blots confirmed an increased NF-kappaBp65 nuclear translocation and a decrease of NF kappaBp65 in the cytoplasm, while no difference in postprandial NF-kappaB nuclear translocation was observed following intake of S-casein or G-casein. CONCLUSION: Postprandial mononuclear NF-kappaB activation after a single meal is independent of the AGE-content of the ingested protein. PMID- 16705549 TI - The combination of increased ovarian volume and follicle number is associated with more severe hyperandrogenism in German women with polycystic ovary syndrome. AB - The prevalence and relevance of polycystic ovaries (PCO) in German women with polycystic ovary syndrome has not been evaluated. This retrospective study included 212 PCOS patients (mean age 28 years) diagnosed by the NIH-criteria and consecutively recruited since 2003. Clinical features including anthropometric variables and the degree of hirsutism, family history, menstrual cyclicity as well as endocrine biochemical parameters were recorded. In addition, 3-h oral glucose tolerance testing for indices of insulin resistance and glucose metabolism was performed in each patient. Transvaginal ultrasound was used to detect polcystic ovaries, defined as the presence of at least one ovary > 10 ml or with at least 12 follicles of 2-9 mm diameter. In this German PCOS cohort, PCO were identified in 166 women (78%). Women with PCO (PXO+) had significantly higher LH/FSH ratios (median 2.1 vs. 1.7) and IGF-1 levels (median 182.5 vs. 160.5 ng/ml) compared to patients without PCO (PCO-). In addition, a significantly higher prevalence of acne (50% vs. 33%) and higher hirsutism scores (median 9 vs. 7) were found in PCO+ patients. Testosterone levels and the free androgen index (FAI) correlated significantly with ovarian volume and the number of ovarian follicles. Also, a subgroup of PCO+ women with a combination of increased ovarian volume and follicle number had higher testosterone levels (median 3.1 vs. 2.1 nmol/l) and FAI (median 7.6 vs. 4.5) compared to women with increased follicle count but normal volume. No differences were found in metabolic parameters or insulin resistance indices. PCO are common finding in German PCOS women. PCO appear to be associated with a more pronounced hyperandrogenemia, especially when both ovarian volume and follicle number are increased. PMID- 16705550 TI - Is it necessary to measure free testosterone to assess hyperandrogenemia in women? The role of calculated free and bioavailable testosterone. AB - Hirsutism in women is defined as excessive facial and/or body terminal hairs showing a masculine distribution; the condition affects approximately 7% of women of reproductive age, and chronic anovulation is a common problem for infertile couples, with a rate of 20-25%. There is a general consensus that these women should be evaluated endocrinologically, as many are found to have an androgen excess (AE) disorder. Free testosterone (FT) is the most prevalent marker in women with androgen excess, but the reference measurement procedures for FT are time-consuming and complex manual procedures that are not routinely practicable in large laboratories. Recently, models have been developed for calculating FT from total testosterone (TT), sex hormone binding globulin (SHBG), and albumin. These calculated values have been found to correlate closely with values estimated using the reference measurement procedures. This study compared measured endocrinological parameters--TT, free testosterone (aFT) by analogue ligand immunoassay method, dihydrotestosterone (DHT), dehydroepiandrosterone sulfate (DHEAS), (SHBG), And calculated parameters--calculated free testosterone (cFT), calculated bioavailable testosterone (cBT), and the free androgen index (FAI) in hirsute women and women with polycystic ovary syndrome (PCOS)--with the values in control individuals. A modified Ferriman-Gallwey score was use to describe the hirsutism pattern. No differences were observed when the measured hormone parameters were compared, while the calculated parameters were significantly increased in women in the hirsutism and PCOS groups in comparison with the values in the control group. Calculate parameters mat be more appropriate markers for assessing hyperandrogenemia in women in comparison with measured values of simple enzyme immuno-assays. These calculated values may be capable of replacing the values estimated using reference measurement procedures, so that time-consuming and complex manual procedures for measuring free testosterone with the reference methods may be dispensable in clinical practice. PMID- 16705551 TI - Hyperprolactinemia of pregnancy is not associated with increased in vivo platelet activity and shortened in vitro bleeding times. AB - The aim of the study is to investigate whether platelet activity is increased by hyperprolactinemia during pregnancy as reflected by beta-thromboglobulin level. Forty-eight healthy, pregnant, and 30 healthy, non-pregnant women were investigated with respect to platelet count, collagen/ADP and collagen/epinephrine closure times, beta-thromboglobulin and prolactin levels. The comparison of the variables between the two groups was made by Mann-Whitney U test. The correlation analyses were performed by Spearman's rank correlation test. Our results revealed that platelet counts, collagen/ADP and collagen/epinephrine closure times and beta-thromboglobulin showed no statistically significant differences between pregnant and non-pregnant women. We found no significant correlation between prolactin and collagen/ADP closure time (r = 0.175), between prolactin and collagen/epinephrine closure time (r = -0.112) and between prolactin and beta-thromboglobulin (r = 0.220) in pregnant women. Our findings suggest that platelet activity is comparable during pregnant and non pregnant states and there is no significant effect of prolactin on platelet function in vivo as reflected by beta-thromboglobulin level. PMID- 16705552 TI - Double germline mutations in the RET Proto-oncogene in MEN 2A and MEN 2B kindreds. AB - Medullary thyroid carcinoma (MTC) is a rare form of thyroid cancer representing about 10% of all thyroid malignancies. It occurs mostly as a sporadic tumor or in association with autosomal dominant inherited cancer syndromes--multiple endocrine neoplasia (MEN) types 2A and 2B and familial MTC. Germline mutations in exons 8, 10, 11, 13, 14, 15 and 16 of the RET proto-oncogene are found in most of the familial cases. There are only a few published data reporting multiple germline mutations in the RET proto-oncogene. We have detected double germline mutations in 2 different exons on the same RET allele in two MEN 2 families. In the MEN 2A family, double germline mutation in exons 10 (Cys620Phe) and 13 (Tyr791Phe) was detected. In the MEN 2B family, beside the classical germline mutation in exon 16 (Met918Thr) a second germline mutation in exon 13 (Tyr791Phe) was found. This study revealed that MEN 2 syndromes can also be caused by double germline mutations in the RET proto-oncogene and these families can be added to small worldwide cohort of families with multiple germline mutations. PMID- 16705553 TI - The principle of regulation in biology--from bone to eating behavior. AB - Cell physiology and molecular biology typically follow a reductionistic approach in science. In the last decade, molecular principles and pathogenetic factors involved in the development of many diseases have been successfully discovered. Therefore, early biological concepts based on systemic and cybernetic thoughts have been largely overshadowed by these more recent molecular and pathogenetic factors. This review highlights discoveries on bone development and hypothalamic controlled feeding and eating behavior with a cybernetic and systemic perspective. Interestingly, ancient ideas on bone development and hypothalamic function are still reasonable considerations to embed new molecular discoveries into a systemic concept of principles organizing nature. PMID- 16705554 TI - A diagnostic dilemma in diagnosing and managing an incidental phaeochromocytoma. AB - Incidental adrenal tumours are a common radiological finding. Phaeochromocytomas, though rare, are very important not to be missed, as the consequences could be fatal. Urinary catecholamines and metanephrines measurement is a commonly used method of screening for phaeochromocytomas. We report a patient who had a phaeochromocytoma presented as an incidental adrenal mass on abdominal CT during investigations for recurrent chest infections. The patient had symptoms suggestive of catecholamines excess and hypertension. Repeated urinary catecholamines measurements, plasma catecholamines, 24-hour urinary catecholamines and 24-hour urinary metanephrines all were normal. MIBG scan showed increased uptake in the adrenal mass. Histological examination of the adrenal mass was consistent with phaeochromocytoma. Removal of the adrenal mass resulted in normalisation of blood pressure and abolishing of the patient's symptoms. This case highlights the difficulty in deciding on further management of patients presenting with an adrenal incidentaloma. This case also highlights the difficulty in diagnosing phaeochromocytoma and represents a reminder that plasma and urine catecholamines measurements could be normal in patients with symptomatic phaeochromocytoma. This case also highlights that MIBG scan could reveal a phaeochromocytoma despite absence of biochemical markers. PMID- 16705555 TI - Late diagnosis of congenital adrenal hyperplasia due to 21-hydroxylase deficiency. AB - Congenital adrenal hyperplasia results from 21-hydroxylase deficiency in more than ninety percent of cases. The classical form of 21-hydroxylase deficiency presents in the neonatal period with virilization or adrenal insufficiency, with or without concurrent salt wasting. We report on a rare case of classic 21 hydroxylase deficiency diagnosed in late adulthood. A 39-year-old male patient presented for workup of infertility. Urologic investigation revealed small testes, bilateral testicular masses, and asthenozoospermia. The patient's steroid metabolism showed markedly increased levels of adrenal androgens, in particular of 17-hydroxyprogesterone amd 21-deoxycortisol. The gas chromatographic-mass spectrometric (GC-MS) urinary steroid profile was dominated by metabolites of 17 hydroxyprogesterone, while the endogenous glucocorticoid production was subnormally low. ACTH levels in plasma were elevated. These hormonal findings were consistent with 21-hydroxylase deficiency. Therapy with dexamethasone was initiated. The CTP21A2 gene analysis revealed the mutation I172N (ATC --> AAC) in exon 4 of allele 1 and a large gene deletion in allele 2. Cases of 21-hydroxylase deficiency diagnosed in late adulthood are rare; however, clinicians should be alert of this possibility. PMID- 16705556 TI - [Amalgam: a risk assessment using a review of the latest literature through 2005]. PMID- 16705558 TI - [Income differences in health and life expectancy--cross-sectional and longitudinal findings of the German Socio-Economic Panel (GSOEP)]. AB - Income is an important determinant of individual standards of living and participation in social life. Data from the German Socio-Economic Panel shows that the level of income also affects a person's health and life expectancy. People's self-assessment of their health and health-related quality of life follow a distribution pattern which can be described as a gradient: the lower the income, the more frequent the impairments to subjective health. Life expectancy statistics also reflect income differences, primarily due to premature deaths among lower-income groups. Clues for explaining income-related differences in health and life expectancy can be found in the results on health-related behaviour and use of the medical system: people in the lower income groups smoke more frequently, are slaker in sports and are less likely to go to a doctor when their health is impaired. Furthermore, work environment and job-specific influences, stress burdens and reactions, social comparison processes and disease induced processes of declining social mobility and social exclusion are discussed as possible explanations. PMID- 16705559 TI - [Prevention and health promotion for men--possibilities of approach to workers. A qualitative study]. AB - Epidemiological data verify the fact that masculinity is a risky life-form from a health point of view. It is difficult if not impossible to reach men, especially those from the lower social classes, with programmes of health promotion and prevention. In this study, based on qualitative interviews with 14 workers, two questions are asked: where do men see the burdens on their health and what possibilities are there for improving their health by prevention and health promotion. The interviewees maintain that they experience the burdens primarily at work whereas they find their resources mainly at home. Well-being is one of the core ideas of their concept of health. When dealing with health they are mainly motivated by their own will. Nevertheless impetus from outside has become more important in dealing with health. When dealing with health there are various masculine role models among the different orientation models. In this case the traditional role of the breadwinner seems to be linked with greater health problems and fewer resources than in an orientation based on a partnership. The looking after of children by men appears to be a contributory factor to the promotion of their health. This fact should be taken into greater consideration than has the case been up to now in the organisation of work. Promotion of health in the workplace with the focus on resources should become a priority as far as improving in the health of workers is concerned. In the medical care system female and male doctors have been able to exercise a greater influence than before. Programmes of symptom oriented prevention have become accepted whereas traditional programmes of health promotion will only have a chance if they take the men's wishes for autonomy and their family ties into consideration. PMID- 16705560 TI - [Motivational interviewing of unemployed acceptance and effects of counselling to improve health-related behaviour]. AB - PURPOSE: Conception, implementation and evaluation of a risk-reducing and health promoting brief intervention addressed to unemployed. METHOD: Control-group-study with repeated measurement. 131 unemployed took part in the intervention group and 95 persons were matched according to age and sex and served as a reference group. The intervention group were recruited via flyers, press articles and several institutions, the reference group via the journal of a health insurance company. The reference group received a questionnaire twice assessing the same health related variables as the intervention group. INTERVENTION: In the intervention group, life style-related variables - eating habits, exercising, smoking, alcohol consumption - were assessed via questionnaire. In a one-hour counselling session based on Motivational Interviewing, participants received an individual feedback on their results of the questionnaire, including a comparison against standards and a review of their personal risks and negative consequences as well as advice and recommendations, while the decision for or against a behaviour change was left to the participant. RESULTS: 85.2 % of the participants of the intervention reported that the counselling met their expectations, and 86.2 % would recommend the intervention to a friend. Statistically significant effects for the intervention group compared to the control group from pre- to post-measurement could be shown for alcohol consumption, eating habits, and exercise (p or =3), for IgM (optical density ratio >2 ), or for both, whereas 1 was negative for IgG and IgM but positive for HEV RNA in serum. The survey identified 220 jaundiced women among the 1133 pregnant women recorded over 3 months (attack rate, 19.4%). A total of 18 deaths were recorded among these jaundiced pregnant women (specific case fatality ratio, 8.2%). CONCLUSIONS: This large epidemic of HEV infection illustrates the dramatic impact of this disease on pregnant women. Timely interventions and a vaccine are urgently needed to prevent mortality in this special group. PMID- 16705572 TI - A large outbreak of hepatitis E among a displaced population in Darfur, Sudan, 2004: the role of water treatment methods. AB - BACKGROUND: The conflict in Darfur, Sudan, was responsible for the displacement of 1.8 million civilians. We investigated a large outbreak of hepatitis E virus (HEV) infection in Mornay camp (78,800 inhabitants) in western Darfur. METHODS: To describe the outbreak, we used clinical and demographic information from cases recorded at the camp between 26 July and 31 December 2004. We conducted a case cohort study and a retrospective cohort study to identify risk factors for clinical and asymptomatic hepatitis E, respectively. We collected stool and serum samples from animals and performed a bacteriological analysis of water samples. Human samples were tested for immunoglobulin G and immunoglobulin M antibody to HEV (for serum samples) and for amplification of the HEV genome (for serum and stool samples). RESULTS: In 6 months, 2621 hepatitis E cases were recorded (attack rate, 3.3%), with a case-fatality rate of 1.7% (45 deaths, 19 of which involved were pregnant women). Risk factors for clinical HEV infection included age of 15-45 years (odds ratio, 2.13; 95% confidence interval, 1.02-4.46) and drinking chlorinated surface water (odds ratio, 2.49; 95% confidence interval, 1.22-5.08). Both factors were also suggestive of increased risk for asymptomatic HEV infection, although this was not found to be statistically significant. HEV RNA was positively identified in serum samples obtained from 2 donkeys. No bacteria were identified from any sample of chlorinated water tested. CONCLUSIONS: Current recommendations to ensure a safe water supply may have been insufficient to inactivate HEV and control this epidemic. This research highlights the need to evaluate current water treatment methods and to identify alternative solutions adapted to complex emergencies. PMID- 16705573 TI - Pubertal development predicts resistance to infection and reinfection with Schistosoma japonicum. AB - BACKGROUND: In communities where Schistosoma species are endemic, the prevalence and intensity of schistosomiasis is disproportionately high among children, compared with adults. This epidemiologic pattern is consistent with either the slow development of resistance or the requirement of host developmental changes for the expression of resistance. METHODS: We enrolled 87 individuals aged 7-18 years who did not have Schistosoma japonicum infection and 641 individuals aged 7 30 years with S. japonicum infection, all of whom reside in 3 villages in Leyte, Philippines. At baseline, S. japonicum infection was assessed by Kato-Katz thick smear stool examination, and the levels of the pubertal hormone dehydroepiandrosterone sulfate (DHEA-S) in serum were determined. Individuals with S. japonicum infection were treated with praziquantel, after which stool examination and DHEA-S level measurement were performed every 3 months for 18 months. RESULTS: In cross-sectional analyses, the intensity of infection among individuals with high DHEA-S levels was 43% lower (28 eggs per g, n = 243), compared with individuals with low DHEA-S levels (50 eggs per g, n = 242), even after adjusting for age, sex, and village (P = .01). Following praziquantel treatment, increased DHEA-S levels were associated with resistance to reinfection (P = .006). The intensity of reinfection among individuals with high DHEA-S levels was 42% lower, compared with individuals with low DHEA-S levels, even after adjusting for age, baseline intensity of S. japonicum infection, village, sex and water contact (P < .001). CONCLUSIONS: Increased DHEA-S levels in serum, a marker for adrenal development, is associated with reduced S. japonicum infection and reinfection, even after adjusting for age and, by proxy, cumulative exposure. These data suggest that an intrinsic property of host pubertal development mediates, in part, the resistance to infection observed in older individuals. PMID- 16705574 TI - New insights into the observed age-specific resistance to reinfection with Schistosoma japonicum. PMID- 16705575 TI - Trends in drug-resistant Mycobacterium tuberculosis in New York City, 1991-2003. AB - BACKGROUND: Two drug-resistance surveys showed a very high prevalence of drug resistance among isolates obtained from patients with tuberculosis in 1991 and 1994 in New York, New York. METHODS: A cross-sectional survey in April 1997 and a survey of incident cases in April-June 2003 were conducted. The trend in the proportion of drug resistance in the 4 surveys was examined separately for prevalent and incident cases. Risk factors for drug resistance in incident cases were also assessed. RESULTS: The number of patients was 251 in the 1997 survey and 217 in the 2003 survey. Among prevalent cases, the percentage of cases with resistance to any antituberculosis drug decreased from 33.5% in 1991 to 23.8% in 1994 and to 21.5% in 1997 (P < .001, by test for trend); cases of multidrug resistant tuberculosis also decreased significantly, from 19% in 1991 to 6.8% in 1997 (P < .001, by test for trend). Among incident cases in the 4 surveys, the decrease in resistance to any antituberculosis drugs was not statistically significant; however, the decrease in multidrug-resistant tuberculosis (from 9% in 1991 to 2.8% in 2003) was statistically significant (P = .002, by test for trend). However, in 2003, a worrisome increase in incident cases of multidrug resistant tuberculosis (an increase of 23%) was seen among previously treated patients with pulmonary tuberculosis not born in the United States. Human immunodeficiency virus infection, a strong predictor for drug resistance in 1991 and 1994, was not associated with drug resistance in subsequent surveys. CONCLUSIONS: Intensive case management, including directly observed therapy, adherence monitoring, and periodic medical review to ensure appropriate treatment for each patient, should be sustained to prevent acquired drug resistance. PMID- 16705576 TI - Drug-resistant tuberculosis in New York City: lessons to remember. PMID- 16705577 TI - Fever in patients with mixed-species malaria. AB - BACKGROUND: Clinical symptoms of mixed-species malaria infections have been variously reported as both less severe and more severe than those of single species infections. METHODS: Oral temperatures were taken from and blood slides were prepared for 2308 adults who presented at outpatient malaria clinics in Tak Province (Thailand) during May-August 1998, May-July 1999, and May-June 2001 with malaria infections diagnosed by 2 expert research microscopists, each of whom was blinded to the other's reports. RESULTS: In each year, temperatures of patients with mixed Plasmodium vivax-Plasmodium falciparum infections were higher than temperatures of patients with P. vivax or P. falciparum infections. In every mixed-species case, P. falciparum parasitemia was higher than P. vivax parasitemia, but patient temperature was not correlated with the parasitemia of either species or with the total parasitemia. CONCLUSIONS: Among adults who self report to malaria clinics in western Thailand, patients with mixed P. vivax-P. falciparum infections have higher fevers than patients with single-species infections, a distinction that cannot be attributed to differences in parasitemia. This observation warrants more detailed investigations, spanning wider ranges of ages and transmission environments. PMID- 16705578 TI - Lactic acidosis in Gabonese children with severe malaria is unrelated to dehydration. AB - BACKGROUND: Hyperlactatemia is an important and common complication of severe malaria. We investigated changes in fluid compartment volumes in patients with severe malaria and control patients with the use of bioimpedence analysis. METHODS: We estimated extracellular water and total body water volumes in a total of 180 children: 56 with severe malaria, 94 with moderate malaria, 24 with respiratory tract infection, and 6 with severe diarrhea. RESULTS: There was a mean (+/-SD) decrease in total body water volume of 17+/-24 mL/kg (or 3% of total body water volume) in patients with severe malaria. This compares with a mean (+/ SD) decrease in total body water volume of 33+/-28 mL/kg (or 6% of total body water volume) in patients with severe diarrhea. There was no increase in extracellular water volume in patients with severe malaria, suggesting no significant intravascular volume depletion in patients with severe malaria. There was no relationship between lactatemia and any changes in fluid compartment volumes. CONCLUSIONS: The changes in fluid volumes that were observed are unlikely to be of physiological significance in the pathophysiology of severe malaria. PMID- 16705579 TI - Safety of long-term oral posaconazole use in the treatment of refractory invasive fungal infections. AB - BACKGROUND: Invasive fungal infections are found most frequently in immunosuppressed and critically ill hospitalized patients. Antifungal therapy is often required for long periods. Safety data from the clinical development program of the triazole antifungal agent, posaconazole, were analyzed. METHODS: A total of 428 patients with refractory invasive fungal infections (n = 362) or febrile neutropenia (n = 66) received posaconazole in 2 phase II/III open-label clinical trials. Also, 109 of these patients received posaconazole therapy for > or = 6 months. Incidences of treatment-emergent, treatment-related, and serious adverse events and abnormal laboratory parameters were recorded during these studies. RESULTS: Treatment-emergent, treatment-related adverse events were reported in 38% of the overall patient population. The most common treatment related adverse events were nausea (8%) and vomiting (6%). Treatment-related serious adverse events occurred in 8% of patients. Low rates of treatment-related corrected QT interval and/or QT interval prolongation (1%) and elevation of hepatic enzymes (2%) were reported as adverse events. Treatment-emergent, treatment-related adverse events occurred at similar rates in patients who received posaconazole therapy for < 6 months and > or = 6 months. CONCLUSIONS: Prolonged posaconazole treatment was associated with a generally favorable safety profile in seriously ill patients with refractory invasive fungal infections. Long-term therapy did not increase the risk of any individual adverse event, and no unique adverse event was observed with longer exposure to posaconazole. PMID- 16705580 TI - Oral supplementation with Lactobacillus casei subspecies rhamnosus prevents enteric colonization by Candida species in preterm neonates: a randomized study. AB - BACKGROUND: Colonization by Candida species is the most important predictor of the development of invasive fungal disease in preterm neonates, and the enteric reservoir is a major site of colonization. We evaluated the effectiveness of an orally supplemented probiotic (Lactobacillus casei subspecies rhamnosus; Dicoflor [Dicofarm spa]; 6 x 10(9) cfu/day) in the prevention of gastrointestinal colonization by Candida species in preterm, very low birth weight (i.e., < 1500 g) neonates during their stay in a neonatal intensive care unit. METHODS: Over a 12-month period, a prospective, randomized, blind, clinical trial that involved 80 preterm neonates with a very low birth weight was conducted in a large tertiary neonatal intensive care unit. During the first 3 days of life, the neonates were randomly assigned to receive either an oral probiotic added to human (maternal or pooled donors') milk (group A) or human milk alone (group B) for 6 weeks or until discharge from the NICU, if the neonate was discharged before 6 weeks. On a weekly basis, specimens obtained from various sites (i.e., oropharyngeal, stool, gastric aspirate, and rectal specimens) were collected from all patients for surveillance culture, to assess the occurrence and intensity of fungal colonization in the gastrointestinal tract. RESULTS: The incidence of fungal enteric colonization (with colonization defined as at least 1 positive culture result for specimens obtained from at least 1 site) was significantly lower in group A than in group B (23.1% vs. 48.8%; relative risk, 0.315 [95% confidence interval, 0.120-0.826]; P = .01). The numbers of fungal isolates obtained from each neonate (P = .005) and from each colonized patient (P = .005) were also lower in group A than in group B. L. casei subspecies rhamnosus was more effective in the subgroup of neonates with a birth weight of 1001-1500 g. There were no changes in the relative proportions of the different Candida strains. No adverse effects potentially associated with the probiotic were recorded. CONCLUSIONS: Orally administered L. casei subspecies rhamnosus significantly reduces the incidence and the intensity of enteric colonization by Candida species among very low birth weight neonates. PMID- 16705581 TI - Management of Epstein-Barr virus (EBV) reactivation after allogeneic stem cell transplantation by simultaneous analysis of EBV DNA load and EBV-specific T cell reconstitution. AB - BACKGROUND: Epstein-Barr virus (EBV) reactivation is a frequent event after allogeneic stem cell transplantation and may progress to life-threatening lymphoproliferative disease (EBV-LPD) in the absence of adequate EBV-specific T cell immunity. Quantification of EBV DNA load in asymptomatic individuals who are at risk is a useful (although not entirely predictive) indicator of progression to EBV-LPD and guide for preemptive treatment with CD20 antibodies. METHODS: With the aim of improving the identification of patients at risk, we retrospectively analyzed, within a cohort of 25 consecutive allogeneic stem cell transplant recipients at risk for EBV-LPD, the pattern of T cell reconstitution during EBV reactivation in all preemptively treated patients (8 patients). RESULTS: In 6 of 8 cases, a significant T cell reconstitution (i.e., a CD3+ T cell count of >300 cells/microL) was documented during EBV reactivation, which included an expansion of EBV-specific memory T cells, as shown by human leukocyte antigen class I tetramer analysis. Additional evidence for the antiviral potential of this T cell reconstitution was obtained prospectively from a cohort of 14 consecutive allogeneic stem cell transplant recipients at risk for EBV-LPD. EBV reactivation occurred in 3 patients. Preemptive treatment was successfully withheld for 2 of these patients in light of concurrent (EBV-specific) T cell recovery. CONCLUSION: We conclude that analysis of the level of (EBV-specific) T cell reconstitution during EBV reactivation is an important second parameter, in addition to quantification of EBV DNA load, that will be instrumental in a more accurate definition of patients at risk for EBV-LPD who, given their immunoincompetence, will be most certainly dependent on preemptive interventions. PMID- 16705582 TI - Paving the way toward a better understanding of viral T cell interactions after stem cell transplantation. PMID- 16705583 TI - Erythema migrans in solid-organ transplant recipients. AB - Six adult solid-organ transplant recipients who had chronic drug-induced immunosuppression and who presented with solitary erythema migrans were treated with antibiotics administered at the same dosage and for the same duration used for the treatment of early, localized Lyme borreliosis in immunocompetent patients. The patients had a smooth course of illness and a favorable outcome but did not develop a measurable borrelial serum antibody response. PMID- 16705584 TI - Infections due to rapidly growing mycobacteria. AB - Rapidly growing mycobacteria, generally of low virulence, are capable of causing a wide spectrum of infections. Increasing reports in the literature, referral center experiences, and data from the Infectious Disease Society of America Emerging Infectious Disease Network suggest that greater numbers of infections are occurring. Epidemiological study is imperative in understanding the true incidence of these infections and preventing disease in vulnerable hosts. Especially problematic is pulmonary infection due to Mycobacterium abscessus, which is difficult to cure. New agents with enhanced activity against this group and other nontuberculous mycobacteria are needed. Here, we focus on the members of the rapidly growing mycobacteria because of their emerging importance in both sporadic infections and outbreak settings. PMID- 16705585 TI - The antimicrobial therapy puzzle: could pharmacokinetic-pharmacodynamic relationships be helpful in addressing the issue of appropriate pneumonia treatment in critically ill patients? AB - Until recently, the in vitro susceptibility of microorganisms was considered the only fundamental aspect for antibiotic efficacy in treating pneumonia. However, the relevance of pharmacokinetic-pharmacodynamic relationships in optimizing drug exposure has been progressively highlighted. Antimicrobial agents were divided into concentration-dependent or time-dependent groups, with the most consistently relevant pharmacodynamic parameters for efficacy being either the ration of the plasma peak concentration to the minimum inhibitory concentration or the time the plasma concentration persists above the minimum inhibitory concentration of the etiological agent, respectively. For the adequate treatment of pneumonia, optimal pharmacodynamic exposure should be ensured also at the infection site. To investigate this, a methodologically correct approach may be to detect drug concentration levels in the epithelial lining fluid and in the alveolar macrophages for extracellular and intracellular pathogens, respectively. From this perspective, the pharmacokinetic factors--only in some instances--support the achievement of optimal exposure during the treatment of pneumonia with fixed standard dosing regimens of antimicrobials; conversely, in other instances, the pharmacokinetic factors suggest the need for an implemented dosage regimen or even the choice of a different drug. PMID- 16705586 TI - Optimizing treatment for HIV-infected South African women exposed to single-dose nevirapine: balancing efficacy and cost. AB - INTRODUCTION: Nevirapine (NVP) resistance may decrease the effectiveness of viral suppression with NVP-based antiretroviral therapy (ART) in women infected with human immunodeficiency virus (HIV) with previous exposure to single-dose NVP. However, the alternative lopinavir-ritonavir-based ART regimen is more expensive. Our objectives were to project the tradeoffs regarding life expectancy, cost, and cost-effectiveness of these ART regimens for NVP-exposed, HIV-infected women in South Africa. METHODS: We developed a simulation model in which NVP-exposed, HIV infected South African women received 1 of 5 treatment strategies: HIV care without ART, NVP-based ART, lopinavir-ritonavir-based ART, NVP-based ART followed by lopinavir-ritonavir-based ART, or lopinavir-ritonavir-based ART followed by NVP-based ART. The prevalence of NVP resistance was 39%; other data were obtained from the published literature. RESULTS: Projected life expectancy was 43.7 months for women who did not receive ART, 77.4 months for women who received a single NVP-based regimen, and 84.5 months for women who received a single lopinavir ritonavir-based regimen. NVP resistance reduced survival time by up to 11.6 months among women who received NVP-based ART. The cost-effectiveness of NVP based ART was $800 (US dollars) per year of life saved, compared with no ART, and the cost-effectiveness of lopinavir-ritonavir-based therapy was $4400 per year of life saved, compared with NVP-based ART. Lopinavir-ritonavir followed by NVP based ART yielded the greatest life expectancy (105.4 months), had a cost effectiveness of $2300 per year of life saved, and, if the efficacy of NVP-based regimens improved >6 months postpartum, further increased survival. CONCLUSIONS: NVP resistance substantially decreased the projected survival time associated with NVP-based ART, and lopinavir-ritonavir-based ART resulted in a superior survival time but at higher cost. A sequential regimen starting with lopinavir ritonavir-based ART followed by NVP-based ART maximized projected survival and was cost effective in South Africa. PMID- 16705587 TI - Schistosomiasis and HIV in rural Zimbabwe: efficacy of treatment of schistosomiasis in individuals with HIV coinfection. AB - BACKGROUND: There is evidence from experimental models that the praziquantel induced clearance of schistosomiasis is dependent on the host's immune response. Consequently, human immunodeficiency virus (HIV)-related immunodeficiency may impair the effect of praziquantel treatment. METHODS: In a prospective cohort study, schistosome-infected subjects who were or were not coinfected with HIV were treated with praziquantel and followed up 3, 6, and 12 months after treatment. Quantitative measures of intensity of schistosomiasis (egg counts and levels of circulating anodic antigen in serum) and immunodeficiency (CD4+ cell counts and viral loads) were collected. RESULTS: Cure rates based on egg counts 3 months after treatment were satisfactory and were similar for HIV-positive individuals (cure rate, 86%) and HIV-negative individuals (cure rate, 85%); the magnitude of decrease in egg count was equal. Cure rates based on circulating anodic antigen levels were much lower than cure rates based on egg counts, with HIV-positive individuals experiencing significantly less clearance of schistosomiasis (cure rate, 31%) than HIV-negative individuals (cure rate, 52%), whereas the magnitude of decrease in circulating anodic antigen was also lower among HIV-positive individuals (P < .01). CONCLUSION: The effect of praziquantel may be limited to affecting the fecundity of adult schistosomes in the immunocompromised host, thus reducing egg excretion while leaving schistosomes metabolically active, as shown by the fact that levels of antigen production are maintained. Special guidelines for treatment of schistosomiasis in HIV-coinfected individuals may need to be developed. PMID- 16705588 TI - Use of efavirenz is not associated with a higher risk of depressive disorders: a substudy of the randomized clinical trial ALIZE-ANRS 099. AB - BACKGROUND: Efavirenz (EFV) is a highly active antiretroviral drug, use of which is associated with frequent (although transient) neurosensorial adverse reactions. Whether the use of EFV is associated with the risk of depression or suicide remains controversial. METHODS: ALIZE-ANRS (Agence Nationale de Recherches sur le SIDA et les Hepatites Virales) 099 was a 48-week randomized trial involving virologically suppressed, human immunodeficiency virus (HIV) infected patients that compared the maintenance of a treatment regimen that contained protease inhibitors (177 subjects) with a switch to a once-daily combination of EFV, didanosine, and emtricitabine (178 subjects). We used the trial's adverse events reporting system and a self-administered Center for Epidemiologic Studies-Depression Scale questionnaire to assess depressive disorders. Determinants were studied using a multivariate proportional hazards model adjusted for antiretroviral treatment, sex, age, HIV risk factor, history of depression, hepatic disorder, alcohol abuse, and HIV-related or non-HIV related events. RESULTS: Thirty cases of depressive disorder (26 cases of depression and 4 suicide attempts) occurred during treatment in 27 patients (12 patients [7%] and 15 patients [8%] in the protease inhibitor-based and EFV-based treatment arms, respectively; P = .56). In the proportional hazards model, only age (hazard ratio, 1.6 per 10 years younger; 95% confidence interval, 1.0-2.6) and a history of depressive disorder (hazard ratio, 5.0; 95% confidence interval, 2.1-12.0) were associated with a risk of depressive disorders. The proportion of depressive patients (24%), as determined on the basis of the Center for Epidemiologic Studies-Depression Scale data, was stable during the follow-up period, without difference between treatment groups. Patients with a history of depressive disorder were more frequently depressed (53%) than were those without such history (22%; P = .03). CONCLUSIONS: The frequency of depressive disorders was high in this population, but the disorders were not related to EFV treatment. Younger age and a history of depression are important determinants for depression and should be considered for early detection and case management. PMID- 16705589 TI - Ad hominem argument unsuitable for animal antibiotics. PMID- 16705590 TI - Treatment of Shiga-like toxin-producing Escherichia coli infection. PMID- 16705591 TI - Enterobacterial repetitive intergenic consensus-polymerase chain reaction for typing of uropathogenic Escherichia coli is not what it seems. PMID- 16705592 TI - Plasmodium falciparum chloroquine-resistance transporter gene detection in imported Plasmodium falciparum malaria cases. PMID- 16705593 TI - A simple algorithm for the diagnosis of AIDS-associated genitourinary tuberculosis. PMID- 16705594 TI - Cytomegalovirus disease in HIV infection: twenty years of a regional population's experience. PMID- 16705595 TI - Brain abscess due to Arcanobacterium haemolyticum after dental extraction. PMID- 16705597 TI - Coagulase-negative staphylococci in diabetic foot osteomyelitis. PMID- 16705598 TI - Efficacy of short-course intramuscular pentamidine isethionate treatment on Old World localized cutaneous leishmaniasis in 2 patients. PMID- 16705599 TI - What can we learn from studies comparing linezolid with vancomycin in neutropenic patients when vancomycin dosages are not optimized? PMID- 16705601 TI - Toward improving quality in cardiovascular imaging: One woman's contribution. PMID- 16705602 TI - Opportunistic bacteria in implant infections. Knowing them to plan their control. PMID- 16705603 TI - Biofilm formation in medical device-related infection. AB - Medical device-associated infections, most frequently caused by coagulase negative staphylococci, especially Staphylococcus epidermidis, are of increasing importance in modern medicine. Regularly, antimicrobial therapy fails without removal of the implanted device. The most important factor in the pathogenesis of medical device-associated staphylococcal infections is the formation of adherent, multilayered bacterial biofilms. There is urgent need for an increased understanding of the functional factors involved in biofilm formation, the regulation of their expression, and the interaction of those potential virulence factors in device related infection with the host. Significant progress has been made in recent years which may ultimately lead to new rational approaches for better preventive, therapeutic, and diagnostic measures. PMID- 16705604 TI - Emerging Staphylococcus species as new pathogens in implant infections. AB - The vast use of prosthetic materials in medicine over the last decades has been accompanied by the appearance of new opportunistic pathogens previously considered incapable of causing infections with significant morbidity and/or mortality. In this regard, the genus Staphylococcus enlisting numerous species usually characterized by a saprophytic habit covers a special role. Apart from Staphylococcus aureus and Staphylococcus epidermidis, well known for their large prevalence in implant-related infections, a number of further staphylococcal species are progressively being indicated for their pathogenic potential. The increasing attention on these opportunistic bacteria is due to an ever growing number of clinical reports, which is also deriving from a more accurate identification of these species with currently available techniques. This synopsis intends to offer an overview on recently emerging coagulase-negative staphylococci (CoNS) as well as coagulase-positive/-variable staphylococci exhibiting distinct traits of virulence, pathogenicity, and epidemiologic impact depending among others on the medical field, the type of prosthetic device and its anatomic location. PMID- 16705605 TI - Advances in in vitro and in vivo models for studying the staphylococcal factors involved in implant infections. AB - Implant infections due to staphylococci are one of the greatest threats facing patients receiving implant devices. For many years researchers have sought to understand the mechanisms involved in the adherence of the bacterium to the implanted device and the formation of the unique structure, the biofilm, which protects the indwelling bacteria from the host defence and renders them resistant to antibiotic treatment. A major goal has been to develop in vitro and in vivo models that adequately reflect the real-life situation. From the simple microtiter plate assay and scanning electron microscopy, tools for studying adherence and biofilm formation have since evolved to include specialised equipment for studying adherence, flow cell systems, real-time analysis of biofilm formation using reporter gene assays both in vitro and in vivo, and a wide variety of animal models. In this article, we discuss advances in the last few years in selected in vitro and in vivo models as well as future developments in the study of adherence and biofilm formation by the staphylococci. PMID- 16705606 TI - Biofilm related infections: is there a place for conservative treatment of port related bloodstream infections? AB - Vascular catheters are the most frequently used indwelling medical devices and have become necessary tools for patients with chronic or critical illness. Surgically or percutaneously placed venous access ports are used to facilitate long-term intravenous therapy. The widespread use of these devices has resulted in a dramatic increase in catheter-related infections. It implies considerable morbidity, occasional mortality, and an increase in medical costs derived from its diagnosis, treatment, and mainly, prolongation of the patient's in-hospital stay. Treatment of such infections is often difficult due to the presence of biofilms on the port inner surface; inside the biofilms, bacteria are less vulnerable to antimicrobial agents. Current diagnostic strategies are suboptimal, and most successful treatment options require removal of the infected device followed by a course of antimicrobial therapy. There are limited data concerning the efficacy of antibiotic treatment of port-related bloodstream infections without catheter removal. PMID- 16705607 TI - Antibiotic resistant pathogenic bacteria and their resistance genes in bacterial biofilms. AB - Biofilm-forming bacteria are ubiquitous in the environment and also include biofilm-forming pathogens. Environmental biofilms may form a reservoir for risk genes and may act as a challenge for human health. Examples of the health relevance of biofilms are the increase in antibiotic resistant bacteria hosted in biofilms in hospital and environment and consequently the interaction of these bacteria with human cells, e.g. in the immune system. Although data concerning the occurrence and spread of resistant bacteria within hospital care units are available, the fate of these bacteria in the environment and especially in the aquatic environment has barely been investigated. Once antibiotic resistant bacteria have entered the environment, a back coupling by ingestion or other possible entry into the host has to be prevented. Therefore a strategy to investigate paths of entry, accumulation and spread of resistant bacteria in environmental compartments has been developed using quantitative determination of genetic resistance determinants. Additionally a bacterial bioassay assessed bioeffectivity thresholds of low antibiotic concentrations. This approach enables an evaluation of the potential of contaminated waters to exert a selection pressure on bacterial communities and thus promote the persistence of resistant organisms. Completed with an indicator system for the identification of sources of multiresistant bacteria a concept for monitoring and evaluation of environmental compartments with respect to their potential of antibiotic resistance dissemination is suggested. PMID- 16705608 TI - Prevalence and antibiotic resistance of 15 minor staphylococcal species colonizing orthopedic implants. AB - Several species belonging to Staphylococcus genus (non Sau/ non Sep species) exhibit increasing abilities as opportunistic pathogens in colonisation of periprosthesis tissues. Here we report on antibiotic resistance of 193 strains, belonging to non Sau/ non Sep species, consecutively collected from orthopedic implant infections in a period of about 40 months. The 193 strains (representing 17% of all staphylococci isolated) were analysed for their antibiotic resistance to 16 different drugs. Five species turned out more prevalent, ranging from 1 to 5%: S. hominis (4.2%), S. haemolyticus (3.7%), S. capitis (2.7%), S. warneri (2.6%), and S. cohnii (1.6%). Among these, the prevalence of antibiotic resistance to penicillins was similar, ranging from 51% to 66%. Conversely, significant differences were observed for all the remaining antibiotics. For S. haemolyticus the resistances to oxacillin and imipenem, the four aminoglycosides and erythromycin were at least twice that of the other three species which were compared. S. warneri was on the contrary the species with the lowest occurrence of resistant strains. Ten species appeared only rarely at the infection sites: S. lugdunensis, S. caprae, S. equorum, S. intermedius, S. xylosus, S. simulans, S. saprophyticus, S. pasteuri, S. sciuri, and S. schleiferi. The behaviours of these species, often resistant to penicillins, were individually analysed. Differences in both the frequencies and the panels of antibiotic resistances observed among the non Sau/ non Sep species: i) suggest that horizontal spreading of resistance factors, if acting, was not sufficient per se to level their bio-diversities; ii) highlight and confirm the worrisome appearance within the Staphylococcus genus of emerging ""new pathogens"", not homogeneous for their virulence and antibiotic resistance prevalence, which deserve to be recognised and treated individually. PMID- 16705609 TI - Virulence factors in enterococcal infections of orthopedic devices. AB - Enterococci are opportunistic pathogens which today represent one of the leading causes of nosocomial infections. We have examined a collection of 52 Enterococcus faecalis isolated from orthopedic infections to determine if they were characterized by a specific pattern of virulence factors. The isolates were evaluated for biofilm formation, presence of genes coding the enterococcal surface protein (esp) and gelatinase (gelE), as well as for gelatinase production. While the rate of esp-positive isolates was comparable to that found among strains from other clinical sources, we found a significantly higher rate of strong biofilm formers and gelatinase producers. Particularly high was the rate of gelE-carrying strains expressing the gene. Data suggest that these two factors in particular may play an important role in enterococcal infections associated with biomaterials. PMID- 16705610 TI - Animal models of osteomyelitis. AB - There are numerous reports in the literature using animal models of osteomyelitis for investigating pathogenesis, diagnosis, and treatment of bone infections. Rabbits, rats, and dogs are commonly used animals, and, less frequently, chickens, guinea pigs, miniature pigs, goats, and sheep. Commonly used bones for creating local osteomyelitis include tibia, femur, and radius, and, less frequently, mandible and spine. When designing a specific model, one should consider which animal and which bone will be used, which route for inoculation (either local injection or systemically through vascular injection), which bacterial species and how many bacteria should be applied, if and what sclerosing agent, foreign body or implant should be employed, and if local trauma is needed. Basic methods of evaluation include clinical observation, radiography, microbiology, and histology. PMID- 16705611 TI - Automated ribotyping to distinguish the different non Sau/ non Sep staphylococcal emerging pathogens in orthopedic implant infections. AB - Several species belonging to Staphylococcus genus, other than Staphylococcus aureus and Staphylococcus epidermidis (non Sau/ non Sep species), exhibit increasing abilities as opportunistic pathogens in the colonisation of periprosthetic tissues. Consequently, the availability of means for accurate identification is crucial to assess the pathogenic characteristics and to clarify clinical relevance of the individual species. Here, 146 clinical staphylococcal isolates belonging to non Sau/ non Sep species from prosthesis-associated orthopedic infections were analyzed by conventional enzymatic galleries and by automated ribotyping. Twelve different species were recognised: S. capitis, S. caprae, S. cohnii, S. equorum, S. haemolyticus, S. hominis, S. lugdunensis, S. pasteuri, S. sciuri, S. simulans, S. warneri, S. xylosus. Ribotype identifications were compared with the phenotypes obtained by the Api 20 Staph system and/or ID 32 Staph system. ID 32 Staph profiles were more consistent with ribotyping results than Api Staph profiles. Across the different staphylococcal species investigated, correct identifications with Api Staph were 45%, while with ID 32 Staph they were 59%. It has, however, to be mentioned that ID 32 Staph was mostly applied to discriminate unmatched ribotyping and Api Staph identifications, thus to a subpopulation of strains with ""atypical"" metabolic profile. Automated ribotyping provided a correct identification for 91% of the isolates. These results confirm automated ribotyping as a convenient rapid technique, still subject to improvements, which will accurately and rapidly recognise the newly emerging staphylococcal pathogens in implant-related orthopedic infections. PMID- 16705612 TI - Biocompatibility of supercritical CO2-treated titanium implants in a rat model. AB - Supercritical phase CO2 is a promising method for sterilizing implantable devices and tissue grafts. The goal of this study is to evaluate the biocompatibility of titanium implants sterilized by supercritical phase CO2 in a rat subcutaneous implantation model. At 5 weeks post implantation titanium implants sterilized by supercritical phase CO2 produce a soft tissue reaction that is comparable to other methods of sterilization (steam autoclave, ultraviolet light radiation, ethylene oxide gas, and radio-frequency glow-discharge), as indicated by the thickness and density of the foreign body capsule, although there were some differences on the capillary density. Overall the soft tissue response to the implants was similar among all methods of sterilization, indicating supercritical phase CO2 treatment did not compromise the biocompatibility of the titanium implant. PMID- 16705613 TI - Antibacterial activity of zinc modified titanium oxide surface. AB - Titanium-based implants are successfully used for various biomedical applications. However, in some cases, e.g. in dental implants, failures due to bacterial colonization are reported. Surface modification is a commonly proposed strategy to prevent infections. In this work, titanium oxide, naturally occurring on the surface of titanium, was modified by promoting the formation of a mixed titanium and zinc oxide, on the basis of the idea that zinc oxide on titanium surface may act as the zinc oxide used in pharmaceutical formulation for its lenitive and antibacterial effects. The present work shows that it is possible to form a mixed titanium and zinc oxide on titanium surfaces, as shown by Scanning Electron Microscopy and XPS analysis. To this end titanium was preactivated by UV on crystalline titanium oxide, both in the anatase form or in the co-presence of anatase and rutile. By performing antibacterial assays, we provide evidence of a significant reduction in the viability of five streptococcal oral strains on titanium oxide surfaces modified with zinc. In conclusion, this type of chemical modification of titanium oxide surfaces with zinc might be considered a new way to reduce the risk of bacterial colonization, increasing the lifetime of dental system applications. PMID- 16705614 TI - Microbially-influenced corrosion: damage to prostheses, delight for bacteria. AB - In natural and man-made environments, microbial communities thrive as biofilms on living (e.g. tissue) and inanimate (e.g. plastic, metal, wood, mineral) surfaces. Biofilms are found in a wide range of aqueous habitats, including physiological fluids. Numerous types of microorganisms are able to colonize catheters, implants, prosthetics, and other medical devices manufactured from different metallic and non-metallic materials dwelling within a human body. The development of biofilm is facilitated by the production of extracellular polymeric substances (EPS). Biofilms formed on surfaces of metallic materials may alter interfacial electrochemical processes, which can lead to increased corrosion of the colonized substratum. Deterioration of metallic materials in the presence of a biofilm is termed biocorrosion or microbially-influenced corrosion (MIC). In the field of biomaterials, ""biocorrosion"" is commonly used when describing the effect of host tissue on the corrosion of implant metals and alloys. Therefore, to avoid confusion, we will here use the term MIC as a reference to biofilm-influenced corrosion. It is important to realise that although most metals are prone to microbial colonization, i.e. to biofouling, this does not imply that they are susceptible to MIC. For example, a metal such as titanium, accumulates biofilm, however, it still demonstrates excellent resistance against MIC. Corrosion is, by definition, an electrochemical process, therefore, electrochemical techniques are frequently employed to determine and measure the rate of abiotic, as well as biologically driven corrosion reactions. This communication addresses the use of electrochemical techniques for monitoring (i) biofilm formation on and (ii) MIC of implant metals and alloys. PMID- 16705615 TI - New surfaces with hydrophilic/hydrophobic characteristics in relation to (no)bioadhesion. AB - The possibility of biosurfaces with high or low adhesiveness for protein, bacteria or eukaryotic cells is discussed. At the interface surface object/biological milieu, biocompatibility, (no) bioadhesion and (no) biocontamination are shown to be correlated with physico-chemical surface characteristics. First consequence is the (no) possibility of biofilm formation. Substrates with low surface energy could interact only with hydrophobic biomolecules. On substrates with high surface energy, a water monolayer spontaneously formed. Modification of a surface by plasma techniques is a way for engineering biomaterials. Plasma techniques are dry processes and more suitable for biomedical applications. In the field of biomaterial medical devices, in hygienic prevention of nosocomial diseases, in food packaging, the use of substrates with a very hydrophilic character may help to prevent the proliferation of cells and bacteria. Such a technique is so efficient that antibiotic molecules are not necessary. Therefore, surface engineering is a tool for modifying and adapting materials to specific biological applications. PMID- 16705616 TI - A glance at the role of exotoxins in opportunistic bacterial infections. AB - The production and the mechanism of action of exotoxins from Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa are presented. The attack to the immune host's defenses is the main virulence factor of opportunistic bacteria in implant infections, favoring the invasion and colonization of compromised periprosthesis tissues. PMID- 16705617 TI - The antifouling activity of some juvenoids on three species of acorn barnacle, Balanus. AB - Analogues of insect juvenile hormones (juvenoids) have been tested for settling inhibition of cyprids from three species of barnacle, Balanus balanoides (L.), Balanus improvisus Darwin and Balanus amphitrite Darwin. Some 3-alkoxypyridine derivatives exhibited strong activity at mg litre(-1) concentrations; 3,7 dimethyloctyl 2-methyl-5-pyridyl ether (8) gave an EC(50) of 0.006 mg litre(-1) when tested on B. balanoides. When compound 8 and similar juvenoids were incorporated into paints, test panels kept in seawater for 8 months were free of barnacles. PMID- 16705618 TI - Metal nitrosyl reactivity: Acetonitrile-promoted insertion of an alkylidene into a nitrosyl ligand with fission of the NO bond. AB - Treatment of the complexes [Re(NO)2(PR3)2][BAr(F)4] (R = Cy, 1 a; R = iPr, 1 b) with phenyldiazomethane gave the cationic benzylidene species [Re{CH(C6H5)}(NO)2(PR3)2][BAr(F)4] (2 a and 2 b) in good yields. Upon reaction of 2 a and 2 b with acetonitrile, the consecutive formation of [Re(N[triple bond]CCH3)(N[triple bond]CPh)(NO)(OC(CH3)=NH)(PR3)][BAr(F)4] (3 a and 3 b) and [Re(NCCH3)(OC{CH3}NH{C6H5})(NO)(PR3)2][BAr(F)4] (4 a and 4 b) was observed. The proposed reaction sequence involves the coupling of coordinated NO, carbene and acetonitrile molecules to yield the (1Z)-N-[imino(phenyl)methyl]ethanimidate ligand. The coupling of the nitrosyl and the benzylidene is anticipated to occur first, forming an oximate species. The subsequent acetonitrile addition can be envisaged as a heteroene reaction of the oximate and the acetonitrile ligand yielding 3 a and 3 b, which in turn can cyclise and undergo a prototropic shift initiated by an internal attack of the ethaneimidate ligand on the benzonitrile moiety to afford 4 a and 4 b. PMID- 16705619 TI - Ethylene oligomerisation and polymerisation with nickel phosphanylenolates bearing electron-withdrawing substituents: Structure-reactivity relationships. AB - Three SHOP-type catalysts, in which the C=C(O) double bond was substituted by electron-withdrawing substituents, [Ni{Ph2PC(R1)=C(R2)O}Ph(PPh3)] (2: R1,R2 = C(Me)=NN(Ph)-; 3: R1 = CO2Et, R2 = Ph; 4: R1 = CO2Et, R2 = CF3), were assessed as ethylene-oligomerisation and -polymerisation catalysts and compared to Keim's complex, [Ni{Ph2PCH=C(Ph)O}Ph(PPh3)] (1). A rationale for the influence of the double-bond substituents of the P,O-chelate unit on the catalytic properties is proposed, on the basis of X-ray diffraction studies, spectroscopic data and DFT B3 LYP calculations. Whatever their relative electron-withdrawing strength, the R1 and R2 substituents induce an increase in activity with respect to catalyst 1. For those systems in which the basicity of the oxygen atom is decreased relative to that of the phosphorus atom, the chain-propagation rate increases with respect to that for catalyst 1. Reduction of the basicity of the P relative to that of the O, however, induces higher chain-termination rates. PMID- 16705621 TI - Structure and intramolecular flexibility of beta-cyclodextrin complex with (-) epigallocatechin gallate in aqueous solvent. AB - The probable structure of the inclusion complex of beta-cyclodextrin (beta-CD) and (-)-epigallocatechin gallate (EGCg) in D2O was investigated using several NMR techniques. EGCg formed a 1:1 complex with beta-CD, in which the A ring and a portion of the C ring of EGCg were included at the head of the phenolic hydroxyl group attached to C7 of EGCg in the beta-CD cavity from the wide secondary hydroxyl group side. In the 1:1 complex with beta-CD, EGCg maintained the conformation in which the B and B' rings of EGCg took pseudoequatorial and pseudoaxial positions with respect to the C ring, respectively. The structure of the inclusion complexes of beta-CD and EGCg obtained from NMR experiments supported those determined from AM1 semiempirical SCF MO calculations well. PMID- 16705622 TI - Inhibition of insulin secretion by cerulenin might be due to impaired glucose metabolism. AB - BACKGROUND: Cerulenin, an inhibitor of protein acylation, has been used as a tool to study the potential role of protein acylation in a variety of activities in different cells, and in stimulus-secretion coupling in pancreatic islets and clonal beta-cells. METHODS: In the present study we investigated its effects on stimulated insulin secretion, glucose metabolism and utilization, oxygen consumption and ATP levels. RESULTS: In isolated rat pancreatic islets, cerulenin pre-treatment (100 microM) inhibited insulin secretion in response to glucose, and to the non-hydrolysable analogue of leucine, aminobicyclo-[2,2,1]heptane-2 carboxylic acid (BCH). These data are in accord with the hypothesis that protein acylation could be involved in the stimulation of insulin secretion. However, we also found that cerulenin profoundly decreased glucose oxidation, glucose utilization, oxygen consumption and ATP levels. Consequently, decreased metabolism provides an alternative mechanism to inhibition of protein acylation that could explain the inhibition of insulin secretion by cerulenin. CONCLUSIONS: Inhibition of insulin secretion by cerulenin can no longer be taken as evidence in favour of a role for protein acylation in the control of insulin release. As protein acylation is known to be involved in the normal functioning of proteins in stimulus-secretion coupling and exocytosis, more direct approaches to understand its role(s) are required. PMID- 16705623 TI - Effects of hypoxia reoxygenation in brain slices from rats with type 1-like diabetes mellitus. AB - BACKGROUND: The aim of this study was to determine whether the brain tissue of type 1 diabetic animals is more susceptible to damage by hypoxia reoxygenation than healthy animals. METHODS: This study used rats with diabetes of 1, 2 and 3 months (N = 15 rats/group). Brain slices were subjected to hypoxia and reoxygenation for 180 min in vitro. We measured oxidative stress (lipid peroxidation, glutathione concentration and enzyme activities related to glutathione), concentration of prostaglandin E(2) (PGE(2)) and nitric oxide (NO) pathway (nitrite + nitrate, activities of constitutive (cNOS) and inducible (iNOS) nitric oxide synthase). As a parameter of cell death we measured the efflux of lactate dehydrogenase (LDH). RESULTS: After reoxygenation LDH activity increased in comparison to nondiabetic animals by 40, 40.6 and 68.9% in animals with diabetes of 1, 2 and 3 months duration, respectively. These changes were accompanied by greater increases in lipid peroxides (25.4, 93.7 and 92.8%). PGE(2) accumulated in significantly larger amounts in diabetic animals (62.5, 85.5 and 114%), and nitrite + nitrate accumulation was significantly greater in rats with diabetes of 2 (40.2%) and 3 months duration (24.0%). iNOS activity increased significantly in all the groups of diabetic animals, with the largest increases in rats with diabetes of 2 (18.6%) and 3 months duration (21.1%). CONCLUSIONS: The biochemical pathways involved in oxidative stress and neuronal death are more sensitive to hypoxia reoxygenation in type 1-like diabetic, as compared to normal, rats. PMID- 16705624 TI - Improved resolution in two-dimensional 1H NMR spectra of peptides by band selective, homonuclear decoupling during both the evolution and acquisition periods: application to characterization of the binding of peptides by heparin. AB - Two-dimensional 1H NMR experiments that achieve band-selective, homonuclear decoupling in both the indirectly detected F1 and directly detected F2 dimensions were used to assign the highly overlapped 1H NMR spectrum of the peptide Ac SRGKARVRAKVKDQTK-NH2, both free in solution and bound to heparin. Band-selective, homonuclear decoupling during the evolution period was achieved using a double pulsed field gradient spin-echo (DPFGSE) with semi-selective shaped pulses; band selective, homonuclear decoupling during the acquisition period was achieved by time-shared semi-selective shaped pulse decoupling. Regular TOCSY, ROESY and NOESY spectra and TOCSY, ROESY and NOESY spectra measured with band-selective, homonuclear decoupling in the evolution (F1) dimension (BASHD-TOCSY, ROESY and NOESY spectra) and with band-selective, homonuclear decoupling in both the F1 and F2 dimensions (D-(or Double)-BASHD-TOCSY, ROESY and NOESY spectra) are reported and compared for the peptide and its heparin complex. Complete assignment of the 1H-NMR spectra of the free and heparin-complexed peptide was achieved with the high resolution of the D-BASHD-TOCSY, ROESY and NOESY spectra. Characterization of the heparin-complexed peptide is of interest because of the ability of the peptide to neutralize the anticoagulant activity of heparin. PMID- 16705625 TI - Evaluation of two simplified 15N-NMR methods for determining micros-ms dynamics of proteins. AB - Two methods for estimating the microsecond-millisecond dynamics in proteins from only two 15N relaxation parameters at one magnetic field strength are investigated. Thus, the chemical exchange contribution, R(ex), to the transversal relaxation rate, which contains the dynamics information, is evaluated by two methods: (i) one in which the R(ex) term is derived from the 15N R1 and R2 relaxation rates alone, and (ii) one in which it is obtained from the transversal dipole-chemical shift anisotropy (CSA) cross-correlation rate, eta(xy), and the R2 rate. Since the R1, R2, and eta(xy) experiments are fast and sensitive, both methods are attractive in studies where large amounts of dynamical information are required. However, both methods are liable to effects that can compromise the estimation of the R(ex) terms. In the R2/R1 method, internal ps-ns dynamics and rotational anisotropy can interfere with the determination of R(ex), while in the R2/eta(xy) method it can be affected by variations in the 15N chemical shift anisotropy. Here, the applicability of the two methods is investigated using plastocyanin from Anabaena variabilis as an example, and the quality of the obtained R(ex) terms is evaluated both theoretically and experimentally. It is found that the R2/R1 method gives reliable R(ex) terms if the protein is relatively rigid and tumbles fast and nearly isotropically in solution, as for instance plastocyanin, and is preferable in such cases. In contrast, the R2/eta(xy) method gives better results if the protein is flexible or highly non spherical and can be used for such proteins, if the sequential variation in the 15N chemical shift anisotropy is negligible. For exchange terms <1 s(-1) neither method is reliable. PMID- 16705626 TI - Toward a better description of gallo-phosphate materials in solid-state NMR: 1D and 2D correlation studies. AB - We show that weak 2J(71Ga-O-31P), typically approximately 12 Hz in GaPO4, can be used to efficiently establish heteronuclear 31P-71Ga correlation using a MAS HMQC experiment in gallo-phosphate materials. The experiment demonstrated for cristobalite GaPO4 is then applied to Ga(PO3)3, where it allows the differentiation of the signature of three different Ga sites overlapping in the 1D spectrum. PMID- 16705627 TI - Upregulation of secretory connective tissue growth factor (CTGF) in keratinocyte fibroblast coculture contributes to keloid pathogenesis. AB - Connective tissue growth factor (CTGF) plays a critical role in keloid pathogenesis by promoting collagen synthesis and deposition. Previous work suggested epithelial-mesenchymal interactions as a plausible factor affecting the expression of various growth factors and cytokines by both the epithelial and dermal mesenchymal cells. The aim of this study is to explore the role of epithelial-mesenchymal interactions in modulating CTGF expression. Immunohistochemistry was employed to check CTGF localization in skin tissue. Western blot assay was performed on total protein extracts from skin tissue, cell lysates and conditioned media to detect the basal/expression levels of CTGF. Study groups were subjected to serum stimulation (fibroblast-single cell culture) and pharmacological inhibitors targeted against mTOR (Rapamycin), Sp1 (WP631 and Mitoxanthrone), Smad3 (SB431542), and PI3K (LY294002). Increased localization of CTGF in the basal layer of keloid epidermis and higher expression of CTGF was observed in the keloid tissue extract. Interestingly, lower basal levels of CTGF was observed in fibroblast cell lysates cocultured with keloid keratinocytes compared to normal keratinocytes, while the conditioned media from the former culture consistently demonstrated a higher expression of secreted CTGF as compared to the latter group. These results demonstrate an important role of epithelial-mesenchymal interactions in the regulation of CTGF expression. Fibroblasts treated with inhibitors against mTOR, Sp1, Smad3, and PI3K demonstrated a reduced expression of CTGF, suggesting these signaling pathways to be important in the regulation of CTGF expression. Thus, revealing the therapeutic potentials for inhibitors that are selective for these factors in controlling CTGF expression in fibrotic conditions. PMID- 16705628 TI - Analysis of pesticides in water by liquid chromatography-tandem mass spectrometric techniques. AB - Pesticide residues continue to be the focus of many environmental studies, and the number of articles describing the development of more advanced, multiresidue analytical methodologies does not decline. The use of liquid chromatography-mass spectrometry based on single quadrupole or ion trap analyzers is consolidated for this purpose. The implementation, in the near future, of more sophisticated mass analyzers, such as triple quadrupole and hybrid quadrupole-time-of-flight is anticipated for routine analysis. This article reviews the various works published so far in the literature for the determination of pesticides and transformation products (TPs) in water by means of liquid chromatography (LC) coupled to tandem mass spectrometry. It discusses the various ionization sources and analyzers used for this purpose, as well as the extraction procedures employed for previous sample preconcentration. Because of the widespread use of triple quadrupole analyzers for the generation of pesticides levels in water using tandem mass spectrometry, a table compiling the transitions monitored for ca. 70 compounds is also included. PMID- 16705629 TI - Parallel imaging. PMID- 16705630 TI - Parallel RF transmission in MRI. AB - Following the development of parallel imaging, parallel transmission describes the use of multiple RF transmit coils. Parallel transmission can be applied to improve RF excitation, in particular, multidimensional, spatially selective RF excitation. For instance, parallel transmission is able to shorten spatially selective RF pulses in two or three dimensions, or to minimize the occurring SAR. One potential major application might be the compensation of patient-induced B(1) inhomogeneities, particularly at high main fields. This paper provides an overview of selected aspects of this new transmission approach. The basic principles of parallel transmission are discussed, initial experimental proofs are described, and the impact of error propagation on coil design for parallel transmission is outlined. PMID- 16705631 TI - An introduction to coil array design for parallel MRI. AB - The basic principles of radiofrequency coil array design for parallel MRI are described from both theoretical and practical perspectives. Because parallel MRI techniques rely on coil array sensitivities to provide spatial information about the sample, a careful choice of array design is essential. The concepts of coil array spatial encoding are first discussed from four qualitative perspectives. These qualitative descriptions include using coil arrays to emulate spatial harmonics, choosing coils with selective sensitivities to aliased pixels, using coil sensitivities with broad k-space reception profiles, and relying on detector coils to provide a set of generalized projections of the sample. This qualitative discussion is followed by a quantitative analysis of coil arrays, which is discussed in terms of the baseline SNR of the received images as well as the noise amplifications (g-factor) in the reconstructed data. The complications encountered during the experimental evaluation of coil array SNR are discussed, and solutions are proposed. A series of specific array designs are reviewed, with an emphasis on the general design considerations that motivate each approach. Finally, a set of special topics is discussed, which reflect issues that have become important, especially as arrays are being designed for more high performance applications of parallel MRI. These topics include concerns about the depth penetration of arrays composed of small elements, the use of adaptive arrays for systems with limited receiver channels, the management of inductive coupling between array elements, and special considerations required at high field strengths. The fundamental limits of spatial encoding using coil arrays are discussed, with a primary emphasis on how the determination of these limits impacts the design of optimized arrays. This review is intended to provide insight into how arrays are currently used for parallel MRI and to place into context the new innovations that are to come. PMID- 16705632 TI - Autocalibrated coil sensitivity estimation for parallel imaging. AB - Parallel imaging has proven to be a robust solution to the problem of acquisition speed in MRI. These methods are based on extracting spatial information from an array of multiple surface coils in order to speed up image acquisition. One of the most essential elements of any parallel imaging method is the information describing the coil sensitivity distribution throughout the sample. This paper covers some of the advanced methods to obtain coil sensitivity-related information, focusing particularly on the class of methods referred to as autocalibrating. These methods all acquire the data for coil sensitivity estimation directly before, during or directly after the reduced data acquisition. After a review of standard methods for coil sensitivity estimation, some of the basic and advanced autocalibrating methods are reviewed, and some example applications shown. PMID- 16705633 TI - Parallel imaging in cardiovascular MRI: methods and applications. AB - Cardiovascular MR imaging (CVMR) has become a valuable modality for the non invasive detection and characterization of cardiovascular diseases. CVMR requires high imaging speed and efficiency, which is fundamentally limited in conventional cardiovascular MRI studies. With the introduction of parallel imaging, alternative means for increasing acquisition speed beyond these limits have become available. In parallel imaging some image data are acquired simultaneously, using RF detector coil sensitivities to encode simultaneous spatial information that complements the information gleaned from sequential application of magnetic field gradients. The resulting improvements in imaging speed can be used in various ways, including shortening long examinations, improving spatial resolution and/or anatomic coverage, improving temporal resolution, enhancing image quality, overcoming physiological constraints, detecting and correcting for physiologic motion, and streamlining work flow. Examples of each of these strategies will be provided in this review. First, basic principles and key concepts of parallel MR are described. Second, practical considerations such as coil array design, coil sensitivity calibrations, customized pulse sequences and tailored imaging parameters are outlined. Next, cardiovascular applications of parallel MR are reviewed, ranging from cardiac anatomical and functional assessment to myocardial perfusion and viability to MR angiography of the coronary arteries and the large vessels. Finally, current trends and future directions in parallel CVMR are considered. PMID- 16705634 TI - Accelerated parallel imaging for functional imaging of the human brain. AB - Accelerated parallel imaging (PI) techniques have recently been applied to functional imaging experiments of the human brain in order to improve the performance of commonly used single-shot techniques like echo-planar imaging (EPI). Potential benefits of PI-fMRI include the reduction of geometrical distortions due to off-resonance signals, the reduction of signal-loss in areas with substantial signal inhomogeneity, increases of the spatial and temporal resolution of the fMRI experiment and reduction of gradient acoustic noise. Although PI generally leads to a substantial decrease in image signal-to-noise ratio (SNR), its effect on the temporal stability of the signal, which ultimately determines fMRI performance, is only partially determined by image SNR. Therefore, the penalty for using PI is generally not as severe as the SNR reduction. The majority of problems related to single-shot techniques become more severe at an increased magnetic field strength, making PI an important tool in achieving the full potential of fMRI at high field. PMID- 16705635 TI - Encoding and reconstruction in parallel MRI. AB - The advent of parallel MRI over recent years has prompted a variety of concepts and techniques for performing parallel imaging. A main distinguishing feature among these is the specific way of posing and solving the problem of image reconstruction from undersampled multiple-coil data. The clearest distinction in this respect is that between k-space and image-domain methods. The present paper reviews the basic reconstruction approaches, aiming to emphasize common principles along with actual differences. To this end the treatment starts with an elaboration of the encoding mechanisms and sampling strategies that define the reconstruction task. Based on these considerations a formal framework is developed that permits the various methods to be viewed as different solutions of one common problem. Besides the distinction between k-space and image-domain approaches, special attention is given to the implications of general vs lattice sampling patterns. The paper closes with remarks concerning noise propagation and control in parallel imaging and an outlook upon key issues to be addressed in the future. PMID- 16705636 TI - Phased array ghost elimination. AB - Parallel imaging may be applied to cancel ghosts caused by a variety of distortion mechanisms, including distortions such as off-resonance or local flow, which are space variant. Phased array combining coefficients may be calculated that null ghost artifacts at known locations based on a constrained optimization, which optimizes SNR subject to the nulling constraint. The resultant phased array ghost elimination (PAGE) technique is similar to the method known as sensitivity encoding (SENSE) used for accelerated imaging; however, in this formulation is applied to full field-of-view (FOV) images. The phased array method for ghost elimination may result in greater flexibility in designing acquisition strategies. For example, in multi-shot EPI applications ghosts are typically mitigated by the use of an interleaved phase encode acquisition order. An alternative strategy is to use a sequential, non-interleaved phase encode order and cancel the resultant ghosts using PAGE parallel imaging. Cancellation of ghosts by means of phased array processing makes sequential, non-interleaved phase encode acquisition order practical, and permits a reduction in repetition time, TR, by eliminating the need for echo-shifting. Sequential, non-interleaved phase encode order has benefits of reduced distortion due to off-resonance, in plane flow and EPI delay misalignment. Furthermore, the use of EPI with PAGE has inherent fat-water separation and has been used to provide off-resonance correction using a technique referred to as lipid elimination with an echo shifting N/2-ghost acquisition (LEENA), and may further generalized using the multi-point Dixon method. Other applications of PAGE include cancelling ghosts which arise due to amplitude or phase variation during the approach to steady state. Parallel imaging requires estimates of the complex coil sensitivities. In vivo estimates may be derived by temporally varying the phase encode ordering to obtain a full k-space dataset in a scheme similar to the autocalibrating TSENSE method. This scheme is a generalization of the UNFOLD method used for removing aliasing in undersampled acquisitions. The more general scheme may be used to modulate each EPI ghost image to a separate temporal frequency as described in this paper. PMID- 16705637 TI - Incoherent artefact correction using PPI. AB - Multiple coils provide extra information about a patient which is frequently used to shorten exam times. This review looks at how the extra information might be used to reduce incoherent artefacts arising from physiological processes such as motion or pulsatile flow. PMID- 16705638 TI - Potential and feasibility of parallel MRI at high field. AB - This survey focuses on the fusion of two major lines of recent progress in MRI methodology: parallel imaging with receiver coil arrays and the transition to high and ultra-high field strength for human applications. As discussed in this paper, combining the two developments has vast potential due to multiple specific synergies. First, parallel acquisition and high field are highly complementary in terms of their individual advantages and downsides. As a consequence, the joint approach generally offers enhanced flexibility in the design of scanning strategies. Second, increasing resonance frequency changes the electrodynamics of the MR signal in such a way that parallel imaging becomes more effective in large objects. The underlying conceptual and theoretical considerations are reviewed in detail. In further sections, technical challenges and practical aspects are discussed. The feasibility of parallel MRI at ultra-high field is illustrated by current results of parallel human MRI at 7 T. PMID- 16705639 TI - Generalized encoding through the use of selective excitation in accelerated parallel MRI. AB - Selective RF excitation is employed in magnetic resonance imaging (MRI) to achieve a variety of effects, such as slice selection. More elaborate transverse magnetization patterns can be realized via tailored RF excitation pulses, useful for example to image any specific region geometry within the field of view, or, to acquire non-Fourier encoded samples of the underlying magnetization distribution. In this manuscript, we review prior work on the combination of selective RF excitation with parallel MRI acquisition techniques. This combination can be used both to advantageously manipulate the numerical conditioning of the reconstruction problem, as well as to compact the information content of the acquired data so as to improve the achievable acceleration rate. With the latter application it is possible to also consider the acceleration provided by parallel imaging alone as a compaction of information content, which in certain cases can be used to reduce the length of the selective excitations. The main contribution of this review is to show how the combination of selective excitation with parallel imaging provides the latter an added flexibility that can be used to either enhance image quality, increase imaging speed, or both. PMID- 16705640 TI - Three-dimensional modeling of cytomegalovirus DNA polymerase and preliminary analysis of drug resistance. AB - Cytomegalovirus (CMV) is the leading cause of congenital infection and a frequent opportunistic agent in immunocompromised hosts such as transplant recipients and AIDS patients. CMV DNA polymerase, a member of the polymerase B family, is the primary target of all available antivirals (ganciclovir, cidofovir, and foscarnet) and certain variations of this enzyme could lead to drug resistance. However, understanding the drug resistance mechanisms at the atomic level is hampered by the lack of its three-dimensional (3D) structure. In the present work, 3D models of two different conformations (closed and open) for CMV DNA polymerase have been built based on the crystal structures of bacteriophage RB69 DNA polymerase (a member of the polymerase B family) by using the 3D-Jury Meta server and the program MODELLER. Most of the variations on CMV DNA polymerase pertinent to ganciclovir/cidofovir and foscarnet resistance can be explained well based on the open and closed conformation models, respectively. These results constitute a first step towards facilitating our understanding of drug resistance mechanisms for CMV and the interpretation of novel viral mutations. PMID- 16705641 TI - Roles of conserved basic amino acid residues and activation mechanism of the hyperthermophilic aspartate racemase at high temperature. AB - X-ray crystallography has revealed two similar alpha/beta domains of the aspartate racemase from the hyperthermophilic archaeon, Pyrococcus horikoshii OT3. The active site is located in the cleft between the two domains where two cysteine residues face each other. This arrangement allows the substrate to enter the cleft and enables the two cysteine residues to act synergistically. However, the distance between their thiolates was estimated to be 9.6 angstroms, which is beyond the distance for cooperative action of them. We examined the molecular mechanism for the racemization reaction of this hyperthermophilic aspartate racemase by mutational analyses and molecular dynamics simulations. The mutational analyses revealed that Arg48 and Lys164 were essential for catalysis in addition to the putative catalytic cysteine residues. The molecular dynamics simulations revealed that the distance between the two active gamma-sulfur atoms of cysteine residues oscillate to periodically become shorter than the predicted cooperative distance at high temperature. In addition, the conformation of Tyr160, which is located at the entrance of the cleft and inhibits the entry of a substrate, changes periodically to open the entrance at 375 K. The opening of the gate is likely to be induced by the motion of the adjacent amino acid, Lys164. The entrance of an aspartate molecule was observed by molecular dynamics (MD) simulations driven by the force of the electrostatic interaction with Arg48, Lys164, and also Asp47. These results provide insights into the roles of amino acid residues at the catalytic site and also the activation mechanism of a hyperthermophilic aspartate racemase at high temperature. PMID- 16705642 TI - Amino acid substitutions affecting protein dynamics in eglin C do not affect heat capacity change upon unfolding. AB - The heat capacity change upon unfolding (deltaC(p)) is a thermodynamic parameter that defines the temperature dependence of the thermodynamic stability of proteins; however, physical basis of the heat capacity change is not completely understood. Although empirical surface area-based calculations can predict heat capacity changes reasonably well, accumulating evidence suggests that changes in hydration of those surfaces is not the only parameter contributing to the observed heat capacity changes upon unfolding. Because packing density in the protein interior is similar to that observed in organic crystals, we hypothesized that changes in protein dynamics resulting in increased rigidity of the protein structure might contribute to the observed heat capacity change upon unfolding. Using differential scanning calorimetry we characterized the thermodynamic behavior of a serine protease inhibitor eglin C and two eglin C variants with altered native state dynamics, as determined by NMR. We found no evidence of changes in deltaC(p) in either of the variants, suggesting that changes in rigidity do not contribute to the heat capacity change upon unfolding in this model system. PMID- 16705643 TI - Management of intrahepatic recurrence after curative treatment of colorectal liver metastases. AB - BACKGROUND: Management of intrahepatic recurrence after complete surgical treatment for colorectal liver metastases is not well defined. The aim of this study was to analyse the survival results of patients who had repeat liver resection for intrahepatic recurrence and to evaluate prognostic indicators for survival. METHODS: Between 1991 and 2005, 55 patients had repeat liver resection for isolated intrahepatic recurrence. The long-term survival results were assessed. Univariable and multivariable analyses were used to identify prognostic indicators for survival after repeat hepatectomy. RESULTS: The median survival was 53 (range 2-97) months and the 5-year survival rate was 49 per cent. In univariable analysis, size of largest initial liver metastasis, margin of initial liver surgery, carcinoembryonic antigen (CEA) level before and after initial liver surgery, liver disease-free survival, margin of repeat liver surgery, operation type of repeat surgery and CEA level before and after repeat surgery were significant prognostic factors. In multivariable analysis, largest initial liver metastasis 4 cm or less and CEA level 5 ng/ml or less after repeat liver surgery were independently associated with improved survival. CONCLUSION: Repeat hepatectomy can achieve an acceptable survival in selected patients with isolated intrahepatic recurrence. PMID- 16705644 TI - Population-based study of the diagnosis and treatment of gastrointestinal stromal tumours. AB - BACKGROUND: The aim of this retrospective population-based study, which was conducted before the introduction of imatinib, was to evaluate the role of surgery in patients with gastrointestinal stromal tumours (GISTs) and clarify which subgroups might benefit from adjuvant treatment. METHODS: Two hundred and fifty-nine patients with clinically detected GISTs were studied. Univariate and multivariate analyses were performed to identify predictors for recurrent disease and survival. RESULTS: Thirty of 48 patients with high-risk GISTs and all of those with overtly malignant tumours developed recurrent tumour after complete (R0) resection. Thirty-four of 38 first recurrences occurred within 36 months of surgery. No recurrence was observed after 72 months. R0 resection, achieved in 48 (80 per cent) of 60 patients with high-risk tumours, was significantly associated with a decreased risk of death from tumour recurrence (P = 0.008). CONCLUSION: Completeness of surgical resection is an independent prognostic factor in patients with high-risk GISTs. A period of adjuvant treatment with imatinib is recommended in patients with high-risk or overtly malignant GISTs who have undergone R0 resection and have a tumour-free interval of less than 6 years. PMID- 16705645 TI - High- and low-resolution mass spectrometry coupled to liquid chromatography as confirmatory methods of anabolic residues in crude meat and infant foods. AB - Within the European Union the use of anabolic steroids for promoting growth and improving meat-to-fat ratio in food-producing animals has been banned since 1988. For the unequivocal identification of hormone residues in a complex matrix such as meat we have developed a rapid, specific and sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method, in combination with a simple extraction procedure based on the matrix solid-phase dispersion (MSPD). The performances of a triple quadrupole (QqQ) and a quadrupole/time-of-flight (QqTOF) were compared: the QqQ mass spectrometer was found to be more sensitive for almost all studied analytes, but the selectivity was superior using the QqTOF system; the full-scan spectra (acquired without losing sensitivity), mass accuracy and resolution of the hybrid instrument enabled a more probatory analyte identification than that obtained selecting two multiple-reaction monitoring (MRM) transitions with a QqQ. Average recoveries ranged from 80 to 100%, and the detection capabilities (CCbetas) were less than 1.06 ppb with the QqQ instrument and less than 5.20 ppb with the QqTOF instrument for the bovine meat, which proved to be the most complex matrix. PMID- 16705646 TI - Utility of porous graphitic carbon stationary phase in quantitative liquid chromatography/tandem mass spectrometry bioanalysis: quantitation of diastereomers in plasma. AB - A major challenge in selecting an appropriate stationary phase for diastereomeric separation is that it is difficult to predict which of the commercially available stationary phases could achieve the required liquid chromatographic (LC) separation. This work describes the selection and evaluation of a porous graphitic carbon (PGC) column coupled with tandem mass spectrometry (MS/MS) for the simultaneous quantitation of an experimental drug candidate (I), its two diastereomeric metabolites (II and III), and its demethylated metabolite (IV) in rat plasma. In addition, we investigated the PGC column for the separation of another drug candidate (VI), its two diastereomeric metabolites (VII and VIII) and its ketone metabolite (IX). The PGC column showed excellent chromatographic resolution for the two diastereomers II and III, as well as for VII and VIII. In contrast, the required resolution for the diastereomers II and III could not be achieved using silica-bonded C(18), C(30), phenyl, perfluorinated, polar embedded and polar end-capped phases. The PGC column showed ruggedness with excellent reproducibility of retention times, peak symmetry and response over a period of more than 400 injections of a plasma acetonitrile-precipitation extract. Excellent accuracy and precision were achieved, with accuracy of 94-108% and intra- and inter-run precision within 9%. This work indicates that PGC is a valuable addition to the repertoire of LC columns used for quantitative LC/MS/MS bioanalysis, especially where the separation and quantitation of diastereomeric analytes is involved. PMID- 16705647 TI - Combined infrared multiphoton dissociation and electron-capture dissociation using co-linear and overlapping beams in Fourier transform ion cyclotron resonance mass spectrometry. AB - A novel set-up for Fourier transform ion cyclotron resonance mass spectrometry (FTICR) is reported for simultaneous infrared multiphoton dissociation (IRMPD) and electron-capture dissociation (ECD). An unmodified electron gun ensures complete, on-axis overlap between the electron and the photon beams. The instrumentation, design and implementation of this novel approach are described. In this configuration the IR beam is directed into the ICR cell using a pneumatically actuated mirror inserted into the ion-optical path. Concept validation was made using different combinations of IRMPD and ECD irradiation events on two standard peptides. The ability to perform efficient IRMPD, ECD and especially simultaneous IRMPD and ECD using lower irradiation times is demonstrated. The increase in primary sequence coverage, with the combined IRMPD and ECD set-up, also increases the confidence in peptide and protein assignments. PMID- 16705648 TI - Detection of 17alpha-hydroxyprogesterone caproate in equine plasma by gas chromatography/tandem mass spectrometry. AB - A method was developed for the analysis of the synthetic progestin 17alpha hydroxyprogesterone caproate in equine plasma following its administration by intramuscular injection. The method employed a reversed-phase solid-phase extraction followed by enol-trimethylsilylation and analysis by gas chromatography/tandem mass spectrometry. The intact ester was detectable in the plasma for up to 2 weeks after a single therapeutic dose, and was found to be stable in equine whole blood for at least 2 months. PMID- 16705649 TI - A statistical framework to discover true associations from multiprotein complex pull-down proteomics data sets. AB - Experimental processes to collect and process proteomics data are increasingly complex, and the computational methods to assess the quality and significance of these data remain unsophisticated. These challenges have led to many biological oversights and computational misconceptions. We developed an empirical Bayes model to analyze multiprotein complex (MPC) proteomics data derived from peptide mass spectrometry detections of purified protein complex pull-down experiments. Using our model and two yeast proteomics data sets, we estimated that there should be an average of about 20 true associations per MPC, almost 10 times as high as was previously estimated. For data sets generated to mimic a real proteome, our model achieved on average 80% sensitivity in detecting true associations, as compared with the 3% sensitivity in previous work, while maintaining a comparable false discovery rate of 0.3%. Cross-examination of our results with protein complexes confirmed by various experimental techniques demonstrates that many true associations that cannot be identified by previous approach are identified by our method. PMID- 16705650 TI - Influence of various cell-detachment solutions on endothelial cells after catheter abrasion for prosthesis colonization prior to implantation. AB - We (1) evaluated the effectiveness of different media on the detachment of endothelial cells (ECs) from a catheter brush (Cragg thrombolytic catheter system) that was previously used for endothelial cell abrasion in 10 cm human umbilical vein (HUV) segments; (2) tested the practicability of endovascular catheter brush abrasion followed by EC detachment from the catheter brush, in vitro culture of harvested ECs, and finally endothelialization of a prosthesis; and (3) analyzed the defect created by the catheter brush in HUV segments after endovascular catheter abrasion. Best results in detachment of ECs from the catheter brush were obtained with a mixture of phosphate-buffered saline + 1% human albumin. EC vitality was time-dependent in the collected HUV segments postdelivery. Harvested EC viability decreased from (26.28 +/- 5.76)% (0-3 h postdelivery) to (17.29 +/- 4.56)% (after 4-8 h). ECs were easily cultured ex vivo within 2-3 weeks; seeded on nitinol stents, they grew to confluency and formed a monolayer on the stent surface (determined by scanning electron microscopy - SEM). Histological and SEM analysis of HUV segments that had undergone previous catheter brush abrasion revealed slight disruption of the intima but intact subintimal layers. Our findings indicate an advantageous method of capturing and culturing primary ECs for gene therapy or for the analysis and diagnosis of certain blood vessel diseases, especially in cases in which endovascular intervention is performed anyway. Moreover, and of high relevance to the biomaterial field, theoretically the procedure could be used to endothelialze a prosthesis ex vivo for implantation into the patient from whom the ECs were harvested, to reduce the inherent thrombogenicity of the prosthesis. PMID- 16705651 TI - Interaction of (-)-epigallocatechin-3-gallate with human serum albumin: fluorescence, fourier transform infrared, circular dichroism, and docking studies. AB - (-)-Epigallocatechin-3-gallate (EGCG), the major constituent of green tea has been reported to prevent many diseases by virtue of its antioxidant properties. The binding of EGCG with human serum albumin (HSA) has been investigated for the first time by using fluorescence, circular dichroism (CD), Fourier transform infrared (FTIR) spectroscopy, and protein-ligand docking. We observed a quenching of fluorescence of HSA in the presence of EGCG. The binding parameters were determined by a Scatchard plot and the results were found to be consistent with those obtained from a modified Stern-Volmer equation. From the thermodynamic parameters calculated according to the van't Hoff equation, the enthalpy change deltaH degrees and entropy change deltaS degrees were found to be -22.59 and 16.23 J/mol K, respectively. These values suggest that apart from an initial hydrophobic association, the complex is held together by van der Waals interactions and hydrogen bonding. Data obtained by fluorescence spectroscopy, CD, and FTIR experiments along with the docking studies suggest that EGCG binds to residues located in subdomains IIa and IIIa of HSA. Specific interactions are observed with residues Trp 214, Arg 218, Gln 221, Asn 295 and Asp 451. We have also looked at changes in the accessible surface area of the interacting residues on binding EGCG for a better understanding of the interaction. PMID- 16705652 TI - Dynamic control of allosteric antagonism of leukocyte function antigen-1 and intercellular adhesion molecule-1 interaction. AB - Leukocyte function associated antigen-1 (LFA-1) plays a critical role in T cell migration and has been recognized as a therapeutic target for immune disorders. Several classes of small molecule antagonists have been developed to block LFA-1 interaction with intercellular adhesion molecule-1 (ICAM-1). Recent structural studies show that the antagonists bind to an allosteric site in the I-domain of LFA-1. However, it is not yet clear how these small molecules work as antagonists since no significant conformational change is observed in the I-domain-antagonist complex structures. Here we present a computational study suggesting how these allosteric antagonists affect the dynamics of the I-domain. The lowest frequency vibrational mode calculated from an LFA-1 I-domain structure shows large scale "coil-down" motion of the C-terminal alpha7 helix, which may lead to the open form of the I-domain. The presence of an allosteric antagonist greatly reduces this motion of the alpha7 helix as well as other parts of the I-domain. Thus, our study suggests that allosteric antagonists work by eliminating breathing motion that leads to the open conformation of the I-domain. PMID- 16705653 TI - N-terminal H3/D3-acetylation for improved high-throughput peptide sequencing by matrix-assisted laser desorption/ionization mass spectrometry with a time-of flight/time-of-flight analyzer. AB - A novel method for peptide sequencing by matrix-assisted laser desorption/ionization mass spectrometry with a time-of-flight/time-of-flight analyzer (MALDI-TOF/TOF) is presented. A stable isotope label introduced in the peptide N-terminus by derivatization, using a 1:1 mixture of acetic anhydride and deuterated acetic anhydride, allows for easy and unambiguous identification of ions belonging either to the N- or the C-terminal ion series in the product ion spectrum, making sequence assignment significantly simplified. The good performance of this technique was shown by successful sequencing of the contents of several peptide maps. A similar approach was recently applied to nanoelectrospray ionization (nanoESI) and nano-liquid chromatography/tandem mass spectrometry (LC/MS/MS). The MALDI-TOF/TOF technique allows for fast, direct sequencing of modified peptides in proteomics samples, and is complementary to the nanoESI and nanoLC/MS/MS approaches. PMID- 16705654 TI - Fixation of nitrogen in an electrospray mass spectrometer. AB - The formation of an iron-nitrogen compound in an electrospray ionization instrument is reported. The iron was released from the stainless steel electrospray needle by acetic acid added to displace trifluoroacetic acid and the nitrogen was present as the drying gas. The product is an iron nitride ion, [N(2)FeOH](1+), m/z 100.9438, and is capable of addition to peptides and proteins. PMID- 16705655 TI - Psychopathy and malingering of psychiatric disorder in criminal defendants. AB - Because the use of deception is an important clinical characteristic of psychopathy, there is intuitive appeal to the idea that psychopathy and malingering are associated. There is, however, very little research showing that psychopaths are more likely to malinger. We classified a sample of criminal defendants as high, moderate, or low in psychopathy, based on PCL-R scores, and compared their scores on indices typically used to detect malingering on the MMPI 2, the PAI and the SIRS. The high psychopathy group scored significantly higher on the MMPI-2 F and F-K; the PAI NIM, and the SIRS, but not the MMPI-2 Fb, F(p), or the PAI RDF or Mal, lending some support for the DSM-IV recommendation that malingering should be considered whenever there is a diagnosis of antisocial personality disorder. Logistic regression analysis (LGA) revealed that Factor I but not Factor II of the PCL-R significantly discriminated malingerers from nonmalingerers with 75% correct classification. However, receiver operating characteristic (ROC) analysis revealed that psychopathy ratings had poor sensitivity and specificity in the detection of malingering. Calculation of the percentages of those exceeding accepted cut-offs on each of the malingering measures revealed that a high percentage of severe psychopaths did not attempt to feign psychiatric disorder. The results suggest that psychopathy is not a clinically useful indictor of malingering. The results are discussed in terms of confirmatory bias and the impact such a bias could have on the evaluation of criminal defendants. PMID- 16705656 TI - Vexatious litigants and unusually persistent complainants and petitioners: from querulous paranoia to querulous behaviour. AB - Querulous paranoia was once of considerable clinical and academic interest in psychiatry. Over the last 40 years, however, it has virtually disappeared from the professional landscape. This decline occurred at the very time that a proliferation of complaint organizations and agencies of accountability were drawing more and more people into asserting their individual rights through the pursuit of claims and grievances. Querulous behaviour, as a result, far from declining, is on the increase, bringing with it suffering for the querulous and disruption to the organizations through which they seek their vision of justice. This article examines querulous behaviour in the vexatious litigant and in abnormally persistent complainants and petitioners. The phenomenological and nosological issues are outlined and the risks of the emergence of threatening and violent behaviour is emphasized. Threats should not be ignored, for a variety of reasons. Approaches to managing querulous behaviour in the courts and the complaint organizations are discussed, together with the utility of individual therapy. Querulous behaviour should once more take its place among the legitimate concerns of mental health professionals. Those caught up in a querulous pursuit of their notion of justice are amenable to management that can ameliorate their suffering and reduce the disruption they create. PMID- 16705657 TI - Genetics and delusional disorder. AB - This article gives an overview of genetic research approaches and their application to delusional disorder. Most studies have been based on small samples and have had other methodological limitations, so it is not clear whether there is a genetic contribution to the aetiology of delusional disorder. It is unlikely that delusional disorder is strongly related genetically to affective disorder or schizophrenia, but more subtle relationships cannot be ruled out. The rarity of multiply affected families prohibits linkage studies and, to date, molecular genetic investigations have been mainly limited to small association studies of dopamine receptor polymorphisms. A range of considerably larger, epidemiologically rigorous studies is required, but the uncommonness and other features of the disorder put strong limitations on the prospects for ascertaining adequate samples. PMID- 16705658 TI - Kidnapping: a criminal profile of persons convicted 1979-2001. AB - Kidnapping has been a neglected crime in criminological research. In fact, there has been a dramatic increase in the yearly numbers of police recorded kidnapping offences in England and Wales in the last 25 years, but this has not been matched by a similar increase in convictions. This study focuses on the official criminal histories of the 7042 males and 545 females who were convicted at least once for kidnapping between 1979 and 2001. Of these, 3.9% of the males and 2.6% of the females had convictions for kidnapping on more than one occasion. We examined two subgroups to ensure long observation periods for prior and subsequent convictions. Of those convicted of kidnapping in 2001, around one-half (51.1%) of the males and around one-third (36.6%) of the females had previous convictions. The previous convictions consist of a wide range of offences, with the most common being theft and violent offences. A 20-year follow-up of those convicted in 1979-81 showed that three in five males and one in three females are subsequently convicted on at least one more occasion for a standard-list offence. An examination of convictions for other offences brought to court at the same time as the kidnap offence enabled a typology of kidnaps to be proposed. Those kidnaps with co-convictions of an acquisitive nature declined over the period, whereas other types, including sexual and violent, showed rises. Changing shifts in the nature of kidnaps have important policy implications. PMID- 16705659 TI - Delusional disorder and delusions: is there a risk of violence in social interactions about the core symptom? AB - Delusional disorders are rare, but psychoses with prominent and persistent delusions are less so. A small but significant association between psychosis and violence is often mediated by delusions in such illnesses. Traditionally, delusions have been viewed as "incorrigible", but there is evidence that they change over time. During development of a scale for measuring delusions, it was found that people who acted violently on their "most important" delusion were more likely to have modified that belief after a mild form of challenge to it. When cognitive-behaviour therapy (CBT) is used for schizophrenia, attempts to modify psychotic symptoms are generally included. Could studies of CBT provide further information about possible risks of social interactions about delusions?In the UK, 2000 people with schizophrenia have been in randomized controlled trials of CBT with a goal of symptom modification. These studies were examined for evidence of violence during the treatment. There was none. Given the period prevalence of violence among people with psychosis, this is surprising. In these studies, however, both challenge to delusions and change in them was minimal and in the context of a safe clinical relationship. Challenge to delusions may, however, occur in a variety of social situations. There are no systematic data on lay challenge to them, but it seems likely that some in the sufferer's social circle will do so vigorously. Relatives, friends, and acquaintances are the people most vulnerable to the most serious violence by someone with psychosis. Study of how people interact in these circumstances and whether their interactions are relevant to modification of delusions would be worthwhile. Could those close to a sufferer learn skills for responding to such pathological beliefs that could be protective against violence, perhaps derived from the principles of CBT? PMID- 16705660 TI - Cervical dystonia in spinocerebellar ataxia type 2. PMID- 16705661 TI - Neuropsychological deficits in Parkinson's disease patients with visual hallucinations. AB - Recent neuropathological and neuroimaging studies suggest the involvement of several temporal regions in Parkinson's disease (PD) patients with visual hallucinations (VH). We examined 24 nondemented PD patients with VH, 21 PD patients without VH, and 21 healthy controls using a battery of tests assessing different aspects of temporal lobe function. PD patients with VH showed poorer performance in language, verbal learning, semantic fluency, and visuoperceptive functions compared to controls and PD patients without VH. Differences in verbal learning and visuoperceptive functions were independent of general cognitive status, disease severity, and depression. We suggest that a wide range of neuropsychological deficits can contribute to the emergence of VH in PD. PMID- 16705662 TI - Commentary on Eyler and Jeste: practical issues in consent capacity and mental disorders. PMID- 16705663 TI - Enhancing the informed consent process: a conceptual overview. AB - Concern about limitations in the ability to give valid informed consent among certain groups of individuals has led to increased interest in defining, measuring, and enhancing consent-related decision-making. In this overview, we summarize issues related to the definition of decision-making capacity, discuss methods that have been used to assess decision-making abilities and other aspects of informed consent, and briefly describe studies that aim to understand variation in different elements of informed consent. We then review strategies that have been used with the intent of improving aspects of the informed consent process. Finally, we provide an outline of areas that are in need of future studies in order to reach the ultimate goal of preserving as much autonomy as possible in at-risk populations, while still achieving valuable research and treatment goals. PMID- 16705664 TI - Pharmacotherapy of delusional disorders in the context of offending and the potential for compulsory treatment. AB - Delusional disorder is an uncommon mental illness with an estimated prevalence of 0.03%. Its low prevalence has likely contributed to the paucity of research interest in this area, leading to substantial gaps in knowledge concerning its treatment and management. In the absence of a robust literature, most clinicians rely on their experience and guidelines for treating schizophrenia when treating patients with delusional disorder. This article reviews the available literature that is specific to the treatment of delusional disorder. In addition, it focuses on specific forensic and medicolegal aspects of managing patients with delusional disorder. PMID- 16705665 TI - Role of lysine versus arginine in enzyme cold-adaptation: modifying lysine to homo-arginine stabilizes the cold-adapted alpha-amylase from Pseudoalteramonas haloplanktis. AB - The cold-adapted alpha-amylase from Pseudoalteromonas haloplanktis (AHA) is a multidomain enzyme capable of reversible unfolding. Cold-adapted proteins, including AHA, have been predicted to be structurally flexible and conformationally unstable as a consequence of a high lysine-to-arginine ratio. In order to examine the role of low arginine content in structural flexibility of AHA, the amino groups of lysine were guanidinated to form homo-arginine (hR), and the structure-function-stability properties of the modified enzyme were analyzed by transverse urea gradient-gel electrophoresis. The extent of modification was monitored by MALDI-TOF-MS, and correlated to changes in activity and stability. Modifying lysine to hR produced a conformationally more stable and less active alpha-amylase. The k(cat) of the modified enzyme decreased with a concomitant increase in deltaH# and decrease in K(m). To interpret the structural basis of the kinetic and thermodynamic properties, the hR residues were modeled in the AHA X-ray structure and compared to the X-ray structure of a thermostable homolog. The experimental properties of the modified AHA were consistent with K106hR forming an intra-Domain B salt bridge to stabilize the active site and decrease the cooperativity of unfolding. Homo-Arg modification also appeared to alter Ca2+ and Cl- binding in the active site. Our results indicate that replacing lysine with hR generates mesophilic-like characteristics in AHA, and provides support for the importance of lysine residues in promoting enzyme cold adaptation. These data were consistent with computational analyses that show that AHA possesses a compositional bias that favors decreased conformational stability and increased flexibility. PMID- 16705666 TI - Atmospheric pressure photoionization-mass spectrometry and atmospheric pressure chemical ionization-mass spectrometry of neurotransmitters. AB - A group of five neurotransmitters with different properties was analyzed using atmospheric pressure photoionization (APPI) and atmospheric pressure chemical ionization-mass spectrometry (APCI-MS). The sensitivity of the techniques for the analytes was tested in six solvents and in positive and negative ion modes. APPI was found to be superior in sensitivity for all the compounds in both positive and negative ion modes. In positive ion mode, water/methanol/formic acid was found to be the best solvent, whereas in negative ion mode, water/methanol/ammonium hydroxide performed best. Detection limits using APPI were between 2.5-250 fmol, depending on the compound. The sensitivity was best for the neurosteroids dehydroepiandrosterone and beta-estradiol, and acetylcholine (LOD 2.5-10 fmol). PMID- 16705667 TI - A validated method for the determination of nicotine, cotinine, trans-3' hydroxycotinine, and norcotinine in human plasma using solid-phase extraction and liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry. AB - A liquid chromatographic-mass spectrometric method for the simultaneous determination of nicotine, cotinine, trans-3'-hydroxycotinine, and norcotinine in human plasma was developed and validated. Analytes and deuterated internal standards were extracted from human plasma using solid-phase extraction and analyzed by liquid chromatography/atmospheric pressure chemical ionization-mass spectrometric detection with selected ion monitoring (SIM). Limits of detection and quantification were 1.0 and 2.5 ng/ml, respectively, for all analytes. Linearity ranged from 2.5 to 500 ng/ml of human plasma using a weighting factor of 1/x; correlation coefficients for the calibration curves were > 0.99. Intra- and inter-assay precision and accuracy were < 15.0%. Recoveries were 108.2-110.8% nicotine, 95.8-108.7% cotinine, 90.5-99.5% trans-3'-hydroxycotinine, and 99.5 109.5% norcotinine. The method was also partially validated in bovine serum, owing to the difficulty of obtaining nicotine-free human plasma for the preparation of calibrators and quality control (QC) samples. This method proved to be robust and accurate for the quantification of nicotine, cotinine, trans-3' hydroxycotinine, and norcotinine in human plasma collected in clinical studies of acute nicotine effects on brain activity and on the development of neonates of maternal smokers. PMID- 16705668 TI - Gender-related difference in altered gene expression of a sterol regulatory element binding protein, SREBP-2, by lead nitrate in rats: correlation with development of hypercholesterolemia. AB - Changes in gene expression levels of hepatic sterol regulatory element binding protein-2 (SREBP-2) and 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) after a single i.v. injection of lead nitrate (LN, 100 micromol kg(-1) body weight) were examined comparatively by real time reverse transcriptase-polymerase chain reaction (RT-PCR) in male and female rats. Significant increases in the gene expression level of SREBP-2, a transcription factor for the HMGR gene, occurred at 6-12 h in male and at 24-36 h in female rats after LN-treatment. The gene expression level of HMGR, a rate-limiting enzyme for cholesterol biosynthesis, significantly increased at 3-48 h in male rats and 12-48 h in female rats. Subsequently, significant increases in the amount of hepatic total cholesterol in male and female rats were also observed at 3-48 h and 24-48 h, respectively. The present findings demonstrate that increases in gene expressions of hepatic SREBP 2 and HMGR and the amount of hepatic total cholesterol by LN occur earlier in male rats than in the females, and that increases in the gene expression level of HMGR and the amount of hepatic total cholesterol occur prior to the increase in the gene expression level of SREBP-2 in either sex of rats. PMID- 16705669 TI - Modulation of different stress pathways after styrene and styrene-7,8-oxide exposure in HepG2 cell line and normal human hepatocytes. AB - Styrene is one of the most important monomers produced worldwide. IARC classified styrene as a possible carcinogen to humans (group 2B). Styrene-7,8-oxide (SO) is the main reactive metabolite of styrene, and it is found to be genotoxic in several in vitro test systems. Styrene and styrene-7,8-oxide (SO) toxicity to HepG2 cells was investigated by evaluating end-points such as heat shock proteins (Hsps), metallothioneins (MT), apoptosis-related proteins, accumulation of styrene within the cells and expression of two isoforms of cytochrome P450. The potential activity of styrene and styrene-7,8-oxide in modulating gene expression was also investigated. The results showed induction of Hsp70, metallothioneins, BclX(S/L) and c-myc expression and a decrease in Bax expression in HepG2 after treatments, confirming that these compounds activated protective mechanisms. Moreover, up-regulation of TGFbeta2 and TGFbetaRIII in HepG2 cells was found after exposure to styrene, while in human primary hepatocytes these genes were down-regulated after both treatments. Finally, it was found that styrene and SO treatments did not induce CYP1A2 and CYP2E1 protein expression. In conclusion, both compounds caused toxic stress in HepG2 cells, with SO being more toxic; in the meantime, a different effect of the two compounds in HepG2 cells and primary human hepatocytes was observed regarding their activity in gene modulation. PMID- 16705670 TI - Interspecies in vitro metabolism of the phosphodiesterase-4 (PDE4) inhibitor L 454,560. AB - L-454,560 is a potent phosphodiesterase 4 (PDE4) inhibitor which was identified as a development candidate for the treatment of asthma and chronic obstructive pulmonary disease (COPD). As part of the discovery of this compound, interspecies in vitro metabolism data was generated using liver microsomes and hepatocytes in order to understand the metabolic fate of the compound. In microsomes, metabolism of the 3-methyl-1,2,4-oxadiazole ring was the predominant pathway observed, including ring cleavage. In rat hepatocytes, hydroxylation of the methyl group on the oxadiazole ring and double-bond isomerization were the most abundant metabolites observed. No major species differences were found in terms of microsomal metabolite profiles. The use of LC with UV and MS detection is highlighted, as well as information from tandem mass spectrometry and NMR. PMID- 16705671 TI - Gas chromatography and mass spectrometry of methoxylated polybrominated diphenyl ethers (MeO-PBDEs). AB - Twenty-six methoxylated polybrominated diphenyl ethers (MeO-PBDEs) were characterized by gas chromatography (GC) on four different GC columns with different lengths and polarities, as well as by mass spectrometry using three ionization techniques, electron ionization (EI), electron capture negative ionization (ECNI) and positive ion chemical ionization (PICI). MeO-PBDE congeners with similar retention times on a nonpolar GC column were separated when analyzed on a polar GC column. EI can be used to determine the position of the methoxy substituent (ortho, meta or para) relative to the diphenyl ether oxygen in the MeO-PBDEs. The PICI ionization technique is shown to be valuable to generate structural information of the MeO-PBDEs, i.e. the degree of bromination on both the methoxy-substituted ring and the entirely brominated phenyl ring can be obtained. This structure information can also be achieved for certain MeO-PBDEs with the methoxy group in ortho position to the diphenyl ether bond in ECNI mode. Like other brominated compounds ECNI is preferable when analyzing environmental samples for quantification of MeO-PBDEs because of the formation of bromide ions, which enables low detection limits. PMID- 16705672 TI - Neuropathologic and neuroinflammatory activities of HIV-1-infected human astrocytes in murine brain. AB - The balance between astrocyte and microglia neuroprotection and neurotoxicity defines the tempo of neuronal dysfunction during HIV-1-associated dementia (HAD). Astrocytes maintain brain homeostasis and respond actively to brain damage by providing functional and nutritive neuronal support. In HAD, low-level, continuous infection of astrocytes occurs, but the functional consequences of this infection are poorly understood. To this end, human fetal astrocytes (HFA) and monocyte-derived macrophages (MDM) were infected with HIV-1DJV and HIV-1NL4-3 (neurotropic and lymphotropic strains respectively) and a pseudotyped Vesicular Stomatitis Virus (VSV/HIV-1NL4-3) prior to intracranial injection into the basal ganglia of severe combined immunodeficient mice. Neuropathological and immunohistochemical comparisons for inflammatory and neurotoxic activities were performed amongst the infected cell types at 7 or 14 days. HIV-1-infected MDM induced significant increases in Mac-1, glial fibrillary acidic protein, ionized calcium-binding adapter molecule 1, and proinflammatory cytokine RNA and/or protein expression when compared with HSV/HIV-1- and HIV-1-infected HFA and sham operated mice. Levels of neuron-specific nuclear protein, microtubule-associated protein 2, and neurofilament antigens were reduced significantly in the brain regions injected with human MDM infected with HIV-1DJV or VSV/HIV-1. We conclude that HIV-1 infection of astrocytes leads to limited neurodegeneration, underscoring the early and active role of macrophage-driven neurotoxicity in disease. PMID- 16705673 TI - Localization of the mouse alpha1A-adrenergic receptor (AR) in the brain: alpha1AAR is expressed in neurons, GABAergic interneurons, and NG2 oligodendrocyte progenitors. AB - alpha(1)-Adrenergic receptors (ARs) are not well defined in the central nervous system. The particular cell types and areas that express these receptors are uncertain because of the lack of high avidity antibodies and selective ligands. We have developed transgenic mice that either systemically overexpress the human alpha(1A)-AR subtype fused with the enhanced green fluorescent protein (EGFP) or express the EGFP protein alone under the control of the mouse alpha(1A)-AR promoter. We confirm our transgenic model against the alpha(1A)-AR knockout mouse, which expresses the LacZ gene in place of the coding region for the alpha(1A)-AR. By using these models, we have now determined cellular localization of the alpha(1A)-AR in the brain, at the protein level. The alpha(1A)-AR or the EGFP protein is expressed prominently in neuronal cells in the cerebral cortex, hippocampus, hypothalamus, midbrain, pontine olivary nuclei, trigeminal nuclei, cerebellum, and spinal cord. The types of neurons were diverse, and the alpha(1A) AR colocalized with markers for glutamic acid decarboxylase (GAD), gamma aminobutyric acid (GABA), and N-methyl-D-aspartate (NMDA) receptors. Recordings from alpha(1A)-AR EGFP-expressing cells in the stratum oriens of the hippocampal CA1 region confirmed that these cells were interneurons. We could not detect expression of the alpha(1A)-AR in mature astrocytes, oligodendrocytes, or cerebral blood vessels, but we could detect the alpha(1A)-AR in oligodendrocyte progenitors. We conclude that the alpha(1A)-AR is abundant in the brain, expressed in various types of neurons, and may regulate the function of oligodendrocyte progenitors, interneurons, GABA, and NMDA receptor containing neurons. PMID- 16705674 TI - Development of the neural retina and its vasculature in the marmoset Callithrix jacchus. AB - The morphological sequence of retinal development in the New World marmoset monkey Callithrix jacchus is similar to previous reports in Macaca and humans. The incipient fovea is present at fetal day (Fd) 100 as the only part of the retina that contains five distinct layers, including a single layer of cone photoreceptors. A foveal pit begins to form at Fd 135 in the center of the foveal avascular zone which is surrounded by a ring of blood vessels (BV) and astrocytes. At birth (Fd 144) the fovea has a single layer of cones over the pit center where the inner retinal layers are thinned but still separated. After birth the fovea rapidly matures so that foveal cone and pit morphology are similar to adult by 4 months. Five distinct layers and the BV plexus in the nerve fiber layer are present to the retinal edge in neonatal marmosets. Near the optic disc BV are sprouting into outer retinal layers at birth and vascularization of the outer retina is completed by 2 to 3 months. Retinal length increases sharply up to Fd 135, but undergoes a quiescent period around birth during which pit formation begins. Length then increases again up to 4mo, followed by a slow increase into adulthood. The postnatal increase is accompanied by a marked thinning of the peripheral retina. The pars plana appears after birth and its length increases at least until 2 years of age. The major difference between marmoset and Macaca is the relative immaturity of the marmoset fovea at birth, and its rapid development after birth. This makes the marmoset a good candidate for neonatal experimental manipulation of retinal and eye development. PMID- 16705675 TI - Olivo-cortico-nuclear localizations within crus I of the cerebellum. AB - Retrograde and anterograde tracers were microinjected into the folia of crus I of the cat cerebellum to investigate spatial localization in olivo-cerebellar and cortico-nuclear projections. The folia were shown to be mainly occupied in rostrocaudal succession by three zones receiving their olivo-cerebellar climbing fiber afferents from parts of, respectively, the dorsal lamella of the principal olive, the ventral lamella of the principal olive, and the rostral half of the medial accessory olive. These zones are presumably parts of the D(2), D(1), and C(2) cerebellar cortical zones, as earlier proposed by Rosina and Provini ([1982] Neuroscience 7:2657-2676). Their respective nuclear target territories were found to be in the rostroventral quadrant of nucleus lateralis, the caudoventral quadrant of nucleus lateralis, and the ventral half of nucleus interpositus posterior. The medial-to-lateral width of each zone was shown to be innervated by different groups of olive cells and to project respectively to medial and lateral parts of the nuclear territory for that zone, consistent with the existence in crus I of olivo-cortico-nuclear microcomplexes (cf. Ito [1984] New York: Raven Press). Parts of the length of each zone located within different folia were also shown to relate to different groups of olive cells and to different regions of the zone's overall nuclear territory. Interfolial localizations, which were heavily overlapping in nature, intersected orthogonally with those for zone width. The fine-grain topography implies that individual microzones exist within each of the zones present within crus I. The results also have implications for the possibility that lateral cerebellar pathways are involved in cognition. PMID- 16705677 TI - Twenty-eight retinas but only twelve eyes: an anatomical analysis of the larval visual system of the diving beetle Thermonectus marmoratus (Coleoptera: Dytiscidae). AB - The larvae of the sunburst diving beetle, Thermonectus marmoratus (Coleoptera: Dytiscidae), are highly efficient visually guided predators. Their visual system consists of a cluster of six stemmata and one eye patch on each side of the head capsule. Histological investigations show that the organization of individual stemmata differs strongly from any eye that has previously been described. Based on general morphology, ultrastructural data, and the presence of actin-rich areas that are typical for rhabdoms, we find that each eye is characterized by several retinas. The most dorsal eye on each side is relatively long and tubular, and we have identified three spatially distinct areas with retinula cells: 1) a band of two rows of rhabdoms along the medial side of the eye tube; 2) a flattened cone shaped region towards the bottom of the tube that is formed by the rhabdoms of retinula cells that are oriented perpendicular to the light path; and 3) two horizontal rows of long rhabdoms parallel to the light path at the base of the tube. A second large eye is organized similarly but lacks the medial band. The remaining four eyes are nearly spherical and each has two distinct retinas. The 12 eyes hence account for a total of 26 retinas, and two further retinas are present in eye patches lacking lenses. Our anatomical findings suggest that this is an example of a visual system in which specific visual tasks are distributed among the eyes, and which relies on a variety of highly specialized retinas. PMID- 16705678 TI - Damage of serotonergic axons and immunolocalization of Hsp27, Hsp72, and Hsp90 molecular chaperones after a single dose of MDMA administration in Dark Agouti rat: temporal, spatial, and cellular patterns. AB - 3,4-Methylenedioxymethamphetamine (MDMA, "ecstasy") causes long-term disturbance of the serotonergic system. We examined the temporal, spatial, and cellular distribution of three molecular chaperones, Hsp27, Hsp72, and Hsp90, 3 and 7 days after treatment with 7.5, 15, and 30 mg/kg single intraperitoneal (i.p.) doses of MDMA in Dark Agouti rat brains. Furthermore, we compared the immunostaining patterns of molecular chaperones with serotonergic axonal-vulnerability evaluated by tryptophan-hydroxylase (TryOH) immunoreactivity and with astroglial-activation detected by GFAP-immunostaining. There was a marked reduction in TryOH immunoreactive axon density after MDMA treatment in all examined areas at both time points. Three days after treatment, a significant dose-dependent increase in Hsp27-immunoreactive protoplasmic astrocytes was found in the cingulate, frontal, occipital, and pyriform cortex, and in the hippocampus CA1. However, there was no increase in astroglial Hsp27-immunoreactivity in the caudate putamen, lateral septal nucleus, or anterior hypothalamus. A significant increase in the GFAP immunostaining density of protoplasmic astrocytes was found only in the hippocampus CA1. In addition, numerous strong Hsp72-immunopositive neurons were found in some brain areas only 3 days after treatment with 30 mg/kg MDMA. Increased Hsp27-immunoreactivity exclusively in the examined cortical areas reveals that Hsp27 is a sensitive marker of astroglial response to the effects of MDMA in these regions of Dark Agouti rat brain and suggests differential responses in astroglial Hsp27-expression between distinct brain areas. The co occurrence of Hsp27 and GFAP response exclusively in the hippocampus CA1 may suggest the particular vulnerability of this region. The presence of strong Hsp72 immunopositive neurons in certain brain areas may reflect additional effects of MDMA on nonserotonergic neurons. PMID- 16705679 TI - Innervation of the paraventricular nucleus of the thalamus from cocaine- and amphetamine-regulated transcript (CART) containing neurons of the hypothalamus. AB - The paraventricular nucleus of the thalamus (PVT) is a midline thalamic nucleus with heavy projections to the nucleus accumbens and other limbic regions. Previous studies have shown that the PVT contains fibers immunoreactive for cocaine- and amphetamine-related transcript (CART). The purpose of the present study was to determine the location of CART neurons innervating the PVT of the rat by using retrograde tracing with cholera toxin B (CTb) combined with immunofluorescence for CTb and CART (amino acid sequence 55-102). Immunohistochemical analysis of CART in the dorsal thalamus showed that the PVT is densely innervated by CART fibers whereas adjacent midline and intralaminar thalamic nuclei are unlabeled. Injections of CTb in the dorsal midline thalamus retrogradely labeled neurons in several areas of the hypothalamus and brainstem which also contained CART neurons. The largest number of double-labeled neurons (CTb/CART) was found in the arcuate nucleus of the hypothalamus. CTb/CART neurons were also found in the lateral hypothalamus, zona incerta, and periventricular hypothalamus. These results indicate that the arcuate nucleus is a major source of CART fibers in the PVT. CART neurons in the arcuate nucleus monitor circulating hormonal signals and may regulate food intake and hypothalamic pituitary-adrenal (HPA) activity. Consequently, CART neurons in the arcuate nucleus may transmit signals to the PVT which in turn may influence limbic regions involved in regulating food intake and the HPA. PMID- 16705680 TI - Development of microglia in the cerebral white matter of the human fetus and infant. AB - Although microglial activation may be an initial beneficial response to a variety of insults, prolonged activation can release toxic substances and lead to cell death. Microglial activation secondary to hypoxia-ischemia and/or infection in immature cerebral white matter is important in the pathogenesis of periventricular leukomalacia (PVL), the major pathological substrate of cerebral palsy in the premature infant. We hypothesize that a transient overexpression in activated microglial density occurs normally in the cerebral white matter of the human fetus during the peak window of vulnerability for PVL. Such an increase could render this region susceptible to insults that cause prolonged microglial activation, as conceptualized in PVL. To examine the developmental profile of microglia in the human fetus and infant brain, immunocytochemistry with microglial specific markers were used in 23 control (non-PVL) cases ranging from 20 to 183 postconceptional (PC) weeks. Tomato lectin, used to identify microglial morphology, revealed that the cerebral white matter of the human fetus and infant is densely populated with intermediate and amoeboid microglia; the latter is indicative of an activated state. Quantitative analysis with CD68 showed increased density of activated microglia in the cerebral white matter of the fetus (<37 PC weeks) relative to the neonate/infant (> or =37 PC weeks) and to the overlying cortex of either age group (P = 0.01). The primary finding of a transient, developmental-dependent overabundance of CD68-activated microglia in the cerebral white matter of the fetus suggests a potential "priming" of this area for diverse brain insults characterized by activation of microglia, particularly PVL. J. PMID- 16705681 TI - Aldosterone-sensitive neurons in the nucleus of the solitary tract: efferent projections. AB - The nucleus of the solitary tract (NTS) contains a subpopulation of neurons that express the enzyme 11-beta-hydroxysteroid dehydrogenase type 2 (HSD2), which makes them uniquely sensitive to aldosterone. These neurons may drive sodium appetite, which is enhanced by aldosterone. Anterograde and retrograde neural tracing techniques were used to reveal the efferent projections of the HSD2 neurons in the rat. First, the anterograde tracer Phaseolus vulgaris leucoagglutinin was used to label axonal projections from the medial NTS. Then, NTS-innervated brain regions were injected with a retrograde tracer, cholera toxin beta subunit, to determine which sites are innervated by the HSD2 neurons. The HSD2 neurons project mainly to the ventrolateral bed nucleus of the stria terminalis (BSTvl), the pre-locus coeruleus (pre-LC), and the inner division of the external lateral parabrachial nucleus (PBel). They also send minor axonal projections to the midbrain ventral tegmental area, lateral and paraventricular hypothalamic nuclei, central nucleus of the amygdala, and periaqueductal gray matter. The HSD2 neurons do not innervate the ventrolateral medulla, a key brainstem autonomic site. Additionally, our tracing experiments confirmed that the BSTvl receives direct axonal projections from the neighboring A2 noradrenergic neurons in the NTS, and from the same pontine sites that receive major inputs from the HSD2 neurons (PBel and pre-LC). The efferent projections of the HSD2 neurons may provide new insights into the brain circuitry responsible for sodium appetite. PMID- 16705682 TI - Degradation of chondroitin sulfate proteoglycans potentiates transplant-mediated axonal remodeling and functional recovery after spinal cord injury in adult rats. AB - Transplantation of growth-permissive cells or tissues was used to bridge a lesion cavity and induce axonal growth in experimental spinal cord injury (SCI). Axonal interactions between host and transplant may be affected by upregulation of inhibitory chondroitin sulfate proteoglycans (CSPGs) following various transplantation strategies. The extent of axonal growth and functional recovery after transplantation of embryonic spinal cord tissue decreases in adult compared to neonatal host. We hypothesized that CSPGs contribute to the decrease in the extent to which transplant supports axonal remodeling and functional recovery. Expression of CSPGs increased after overhemisection SCI in adult rats but not in neonates. Embryonic spinal cord transplant was surrounded by CSPGs deposited in host cord, and the interface between host and transplant seemed to contain a large amount of CSPGs. Intrathecally delivered chondroitinase ABC (C'ase) improved recovery of distal forelimb usage and skilled motor behavior after C4 overhemisection injury and transplantation in adults. This behavioral recovery was accompanied by an increased amount of raphespinal axons growing into the transplant, and raphespinal innervation to the cervical motor region was promoted by C'ase plus transplant. Moreover, C'ase increased the number of transplanted neurons that grew axons to the host cervical enlargement, suggesting that degradation of CSPGs supports remodeling not only of host axons but also axons from transplanted neurons. Our results suggest that CSPGs constitute an inhibitory barrier to prevent axonal interactions between host and transplant in adults, and degradation of the inhibitory barrier can potentiate transplant mediated axonal remodeling and functional recovery after SCI. PMID- 16705683 TI - Capillary morphogenesis gene (CMG)-1 is among the genes differentially expressed in mouse male germ line stem cells and embryonic stem cells. AB - We recently established a technique to expand male germ line stem (GS) cells in long-term culture without losing their spermatogenic capacity. To gain insight into the genetic program of these cells, we compared the mRNA expression profile of GS cells with that of embryonic stem (ES) cells using DNA microarrays. We found 79 genes that were upregulated in GS cells compared to ES cells, including synaptonemal complex protein-1, deleted in azoospermia-like, ubiquitin conjugating enzyme E2B, and ubiquitin carboxy-terminal hydrolase L1, all of which are functionally important for spermatogenesis. In addition, we identified a cDNA encoding the mouse ortholog of capillary morphogenesis gene (CMG)-1. CMG-1 transcripts were predominantly produced in spermatogonia and spermatocytes in mouse testis. When CMG-1 expression was attenuated in a mouse spermatocyte derived cell line, GC-2spd(ts), by a target-specific short interfering RNA, the morphology of the cells was changed and the expression of cyclin D2 was abrogated. A reporter assay using a genomic region upstream of the mouse cyclin D2 gene revealed that this downmodulation occurs at the transcriptional level. We detected FLAG-tagged CMG-1 protein in the nuclei of transfected COS7 cells, suggesting that CMG-1 may play a unique role in the transcriptional regulation of the cyclin D2 gene. The upregulated GS genes identified in this study will provide useful information for the future investigation of spermatogonial stem cells and the early phase of male germ cell differentiation. PMID- 16705684 TI - Association between amantadine and the onset of dementia in Parkinson's disease. AB - The objective of this study is to compare the occurrence of dementia among Parkinson's disease (PD) patients treated with amantadine (AM group) with those never exposed to it (NoAM group). PD dementia shares neuroanatomical and biochemical similarities with Alzheimer's disease (AD). Memantine, an N-methyl-D aspartate (NMDA) receptor antagonist has been shown to be beneficial in AD. Memantine is a dimethyl derivative of amantadine, which also possesses NMDA receptor blocking properties. We hypothesized that amantadine could have a beneficial effect on the occurrence of PD dementia. PD patients attending the Movement Disorders Clinics in Hillel Yaffe, Asaf Harofe Medical Centers (Israel) and Pisa (Italy) were included. Taking the onset of dementia as the endpoint, survival curves for AM and NoAM patients were estimated by the Kaplan-Meier method. The study population consisted of 593 patients (age, 69.5 +/- 9.9 years; PD duration, 9.2 +/- 6.0 years; 263 patients (44%) amantadine treated). The endpoint of dementia was reached by 116 patients (20%). PD duration until dementia was significantly longer for AM patients (9.1 +/- 5.7 years) than for NoAM patients (5.9 +/- 4.6 years, P = 0.006). The duration of amantadine exposure positively correlated with PD duration until dementia (P = 0.0001). Survival analysis, taking dementia onset as endpoint, showed slower mental decline in AM patients (Log rank P = 0.0049, Wilcoxon P = 0.0024). Mini-Mental State Examination scores were significantly higher for AM patients than for the NoAM group (P = 0.01). Age of PD onset also significantly influenced the duration of PD until dementia. Amantadine use may delay the onset of dementia in PD patients and may attenuate its severity. PMID- 16705685 TI - Validation of the "L-DOPA test" for diagnosis of restless legs syndrome. AB - We developed and validated a standardized test procedure to evaluate the accuracy of the supportive diagnostic criterion "response to dopaminergic treatment" in restless legs syndrome (RLS). Forty-eight patients who fulfilled at least three of the four essential criteria for RLS, thus including uncertain clinical cases for a nonexpert, were recruited. Patients received a preliminary diagnosis of RLS or non-RLS. All patients underwent a polysomnography (PSG) and were then asked to perform the diagnostic L-DOPA test at home, which consisted in the application of one single dose of 100/25 mg L-DOPA/benserazide and a subsequent observational period of 2 hours. Before, and in 15-minute intervals after, drug intake, the patients rated the severity of the "symptoms in the legs" and the "urge to move the legs" using a 100-mm visual analogue scale. Considering a 50% improvement as a positive test result, we found a sensitivity of 88% ("symptoms in the legs") and 80% ("urge to move the legs") with a specificity of 100% for both test items. A rate of 90% or 83% of all patients could be correctly diagnosed by the L-DOPA test. Both scales were able to predict the response to dopaminergic agents in the subsequent course of the treatment by 100%. The periodic leg movements arousal index as assessed by polysomnography was less appropriate for the prediction of the correct diagnosis. We recommend the L-DOPA test for diagnostic decision making in all patients with an unclear RLS diagnosis according to the essential diagnostic criteria of the International RLS Study Group. PMID- 16705686 TI - Surface electromyography shows increased mirroring in Parkinson's disease patients without overt mirror movements. AB - Patients with Parkinson's disease (PD) may present mirror movements (MM). Transcranial magnetic stimulation data indicate that these movements reflect an abnormal enhancement of the "physiological mirroring" that can be observed in healthy adults during complex and effortful tasks. It was hypothesized that, in PD, enhanced mirroring is caused by a failure of basal ganglia output to support the cortical network that is responsible for the execution of strictly unimanual movements. If so, it is likely that subtle alterations of voluntary unimanual motor control are also present in PD patients without overt MM. We tested this hypothesis by using surface electromyographic (EMG) techniques in 12 mildly to moderately affected PD patients without overt MM, and in 2 control groups (12 age matched and 10 young healthy volunteers). Subjects performed unilateral phasic thumb abduction during a sustained tonic contraction of the opposite abductor pollicis brevis. All patients were tested on dopaminergic therapy. On a separate day, 7 of 12 patients were re-tested after withdrawal of medication. During this task, involuntary mirror-like increase in surface EMG of the tonically abducting thumb was significantly larger in PD patients than in age-matched or young healthy volunteers. Off therapy, mirroring was slightly greater than on medication, although this difference was not significant. Our findings suggest that dysfunction of unimanual motor control is a general feature of PD. It is likely that this deficient movement lateralization contributes to an impairment of nonsymmetrical bimanual movements in PD. PMID- 16705687 TI - Methylphenidate fails to improve gait and muscle tone in patients with sporadic and hereditary spastic paraplegia. AB - Based on its action on multiple neurotransmitters, including dopamine, methylphenidate (MPH) is of growing interest as a possible treatment option for several movement disorders. Of special interest are diseases that share gait disturbance and cognitive decline. Based on a single case observation in a patient with hereditary spastic spinal paraplegia (HSP) in which gait was improved with MPH, we performed an open-label study with a longitudinal follow-up in 22 patients with HSP and its sporadic form (SSP). The patients were treated for 6 months with 60 mg of MPH per day. Computerized gait analysis and different scores were performed at baseline, after 6 weeks, and after 6 months of treatment. Although at 6 weeks, the gait velocity was somewhat improved, the drug failed to show any effect on other gait parameters and had no beneficial effect at all after 6 months. Although MPH is of interest for several movement disorders, our study did not show a beneficial effect. PMID- 16705688 TI - Novel trihydroxamate-containing peptides: design, synthesis, and metal coordination. AB - Novel trihydroxamate-containing peptides were designed to mimic desferrioxamine (Desferal(R), DFO, a naturally occurring siderophore) but possess distinct conformational restrictions and varied lipophilicity to probe structure vs. metal coordination. The synthesis was performed via fragment condensation of hydroxamate-containing oligopeptides such as Fmoc-Leu- Psi[CON(OBz)]-Phe-Ala-Pro OH and H-Leu-Psi[CON(OBz)]-Phe-Ala-Pro-OBu(t) (Fmoc: 9-fluor enylmethoxycarbonyl; OBz: benzyl; OBu(t): tert-butyl) either in solution or on a solid support. The metal-binding properties were studied by electrospray ionization-mass spectroscopy (ESI-MS), ultraviolet (UV)-visible spectroscopy, and (1)H nuclear magnetic resonance (NMR). Similar to the dihydroxamate analogs previously explored [Biopolymers (Peptide Science), 2003, Vol. 71, pp. 489-515], the compounds with three hydroxamates arrayed at 10-atom intervals, i.e., H-[Leu Psi[CON(OH)]-Phe-Ala-Pro](3)-OH (P1), cyclo[Leu-Psi[CON(OH)]-Phe-Ala-Pro](3) (P2), and H-[Leu-Psi(CONOH)-Phe-Ala-Pro](2)-Leu-NHOH (P7), exhibited high affinities for intramolecular coordination with Fe(III) and Ga(III). As expected, both P1 and P2 showed higher relative Fe(III)-binding affinities than the corresponding dihydroxamate-containing peptide analogs (P11 and P12). Even though both P1 and P2 did not compete with DFO in the relative metal-binding affinity in both solution and gas phases, P1, P2, and DFO exhibited similar relative binding selectivities to 11 different metal ions including Fe(III), Fe(II), Al(III), Ga(III), In(III), Zn(II), Cu(II), Co(II), Ni(II), Gd(III), and Mn(II). Compared to the other metal ions, they had higher relative binding affinities with Fe(III), Fe(II), Al(III), Ga(III), and In(III). The decreased metal-binding affinities of P1 and P2 in comparison with DFO suggested the conformational restrictions of their backbones perturb their three hydroxamate groups from optimal hexadentate orientations for metal coordination. As detected by ESI-MS, P2 was distinguished from both P1 and DFO by solvation of its Ga(III) and Fe(III) complexes (such as acetonitrile or water), thereby stabilizing the resulting complexes in the gas phase. Noteworthy, P2 led to 69% death rate in Hela cells at a concentration of 50 microM, exhibiting higher cytotoxicity than DFO in vitro despite its much lower affinity for iron. This enhanced toxicity may simply reflect the increased lipophilicity of the cyclic trihydroxamate (P2) together with the improvements in its cell penetration, and/or subsequent intracellular molecular recognition of both side chains and hydroxamate groups. The cytotoxicity was significantly suppressed by precoordination with Ga(III) or Fe(III), suggesting a mechanism of toxicity via sequestration of essential metal ions as well as the importance of curbing the metal coordination before targeting. The potential of such siderophore-mimicking peptides in oncology needs further exploration. PMID- 16705689 TI - Alterations in surface geometry in retrieved polyethylene glenoid component. AB - This study characterized and quantified the changes found in retrieved glenoid polyethylene components found at revision total shoulder arthroplasty (TSA). Twenty components obtained at revision TSA were evaluated, all from a system (Global, DePuy, Warsaw, IN) with a glenoid radius of curvature 3 mm greater than that of the humeral head. Laser surface scanning provided three-dimensional analysis of the surface of the glenoid component. Scans of unused components of similar sizes enabled determination of the changes occurring after implantation. Alterations in radius of curvature were noted in every glenoid. All showed loss of the balance stability angle (BSA, the maximal angle that the net humeral joint reaction force can make with the glenoid center line before the humeral head would dislocate) of at least 5 degrees in one or more directions. Increase in BSA in one direction was seen in 11 of the components. In five of these, the increase was associated with a reduction of the local radius to match that of the humeral component. Glenoid surface morphology and stability can be changed by in vivo use. While correlation with clinical instability in the patients from whom the implants were obtained was not possible, many of the observed changes in surface morphology are of sufficient magnitude to compromise the contribution of the glenoid surface to shoulder stability. Three patterns of wear were identified: "humeral" that showed loss of the mismatch between the humeral and glenoid radii of curvature (5 of 20 components), "diffuse" that showed broad surface irregularity (18 of 20), and "rim" wear with loss of the polyethylene rim of the component (14 of 20). More than one type of wear was possible within a single glenoid. PMID- 16705690 TI - Prolonged down regulation of specific gene expression in nucleus pulposus cell mediated by RNA interference in vitro. AB - To investigate the efficacies and the longevity of RNA interference in nucleus pulposus cells from rat and human, two reporter luciferase plasmids (Firefly and Renilla) were used. These plasmids were cotransfected with siRNA targeting Firefly luciferase to the nucleus pulposus cells extracted from Sprague Dawley rats and scoliosis patients. The inhibitory effects were evaluated by dual luciferase assay for 3 weeks. Proliferation activity of fibroblast-like cells extracted from the subcutaneous tissue of Sprague Dawley rats and the nucleus pulposus cells were measured by proliferation assay (WST-8 assay) every 2 days after plating. The expression of Firefly luciferase was drastically inhibited both in rats (94.7%) and in humans (93.7%). The inhibitory effects were maintained for 2 weeks and had disappeared completely by 3 weeks. The proliferation activity of nucleus pulposus cells was significantly lower than fibroblast-like cells. We have shown, for the first time, siRNA-mediated gene silencing in rat and human disc cells for a relatively sustained period, probably due to the stability of the nucleus pulposus cells in terms of cell proliferation. The demonstration of this study may allow further exploration of the use of siRNA for scientific research and the treatment of disc degenerative diseases. PMID- 16705691 TI - Mutational spectrum of type I collagen genes in Korean patients with osteogenesis imperfecta. AB - Mutations in the type I collagen genes COL1A1 and COL1A2 are responsible for the dominantly inherited connective tissue disorder osteogenesis imperfecta (OI). The severity of OI is diverse, ranging from perinatal lethality to a very mild phenotype that is characterized by normal stature and the absence of deformities. Although there have been several studies on the mutational spectra of COL1A1 and/or COL1A2 in Western populations, very few cases have been reported from Asia. In this study, we investigated 67 unrelated Korean probands with OI and used nucleotide sequence analysis to detect COL1A1 and COL1A2 mutations. Thirty five different mutations were identified in the two genes, including 24 novel mutations. Among the 35 kinds of detected mutations, 15 were glycine substitutions (seven in COL1A1 and eight in COL1A2), one was a nonsense mutation, four were frameshift mutations in COL1A1, three were in-frame duplications in COL1A2, and 12 were splice site mutations (seven in COL1A1 and five in COL1A2). Until now, mutations in the COL1A1 and COL1A2 genes known to cause OI were unique and rarely repeated in other families. Interestingly, the c.982G>A (p.Gly328Ser) mutation in COL1A2 was found recurrently and was the causative mutation in five independent OI probands. Haplotype analysis of the COL1A2 gene revealed that four probands from five independent OI probands with c.982G>A (p.Gly328Ser) had a common haplotype. Our clinical data showed the heterogeneity even within a specific genotype, which suggested the complex expression of this disease. PMID- 16705692 TI - Distribution of ENG and ACVRL1 (ALK1) mutations in French HHT patients. AB - Hereditary Hemorrhagic Telangiectasia (HHT) is an autosomal dominant disease characterized by arteriovenous malformations and resulting from mutations in two major genes: ENG and ACVRL1. The aim of the present study was to estimate the prevalence of the mutations of ENG and ACVRL1 in HHT, based on the largest series of patients reported so far, recruited through a national network. We previously reported the first mutation screening of both genes, in French HHT patients, using heteroduplex analysis. This previous study, bringing 60 novel mutations, provided a significant improvement to the knowledge of molecular pathology in HHT. However, 32% (n=48) of the patients with a confirmed clinical diagnosis remained without mutation. In these patients, we performed an extensive molecular analysis that included the sequencing of the whole coding sequence, the search for large rearrangements, and screening of the potential 5' regulatory regions. Additionally, due to the lack of large pedigrees suitable for linkage analysis, and since SMAD4 germline mutations have been reported in families with combined HHT and juvenile polyposis, we screened this gene and five other genes involved in the TGF-beta/BMP pathway in the patients without mutation of ENG or ACVRL1. Only a novel SMAD1 non-conservative substitution was found in one patient, changing a poorly conserved methionine to an isoleucin. Twenty-three mutations were found in ACVRL1 and 8 in ENG (including a duplication of exons 4 to 8 and deletions of exons 1 to 3 and 9 to 14). Our results, combined with our previous data, increase the mutation rate to 88% (n=119/136) in French patients with a confirmed clinical diagnosis. Our results also emphasize the higher prevalence of large insertions/deletions in ENG and the predominance of ACVRL1 over ENG mutations. PMID- 16705693 TI - Ultrasound stimulation of types I and III collagen expression of tendon cell and upregulation of transforming growth factor beta. AB - Traumatic tendon injuries are commonly treated with ultrasound. However, previous research has not examined the molecular mechanism of this therapeutic effect on collagen synthesis of tendon cells. This study was designed to determine the effect of ultrasound on the expression of type I and type III collagen of tendon cells intrinsic to rat Achilles tendon. Whether a correlation exits between this effect and the expression of transforming growth factor beta (TGF-beta), which enhances collagen synthesis, was also investigated. Tendon cells after ultrasound treatment and protein expression of types I and III collagen were determined by immunocytochemistry. The mRNA expressions of alpha1(I) procollagen, alpha1(III) procollagen, and TGF-beta were determined by reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, the concentration of TGF-beta in conditioned medium was evaluated by enzyme-linked immunosorbent assay (ELISA). Immunocytochemical staining revealed that ultrasound-treated tendon cells were stained more strongly for types I and III collagen than were control cells. Upregulation of procollagen alpha1(I) gene, procollagen alpha1(III) gene, and TGF beta at the mRNA level was confirmed by RT-PCR. A dose-dependent increase in the concentration of TGF-beta in conditioned medium obtained from cells treated with ultrasound was demonstrated by ELISA assay (p = 0.043). In conclusion, ultrasound stimulates the expression of type I and type III collagen in a process that is likely mediated by the upregulation of TGF-beta. PMID- 16705694 TI - Changes in perlecan during chondrocyte differentiation in the fetal bovine rib growth plate. AB - Perlecan is a heparan sulfate proteoglycan present in the growth plate and essential for endochondral ossification. We evaluated the synthesis and structure of perlecan in the different zones of the growth plate. The growth plates from fetal bovine ribs were isolated and sequentially sliced into 1-mm sections containing the hypertrophic zone, lower proliferative zone, upper proliferative zone, intermediate zone, and resting zone, respectively. The slices were then either incubated in culture medium with 35SO4 to measure total sulfated proteoglycan synthesis and perlecan synthesis, extracted for perlecan core protein analysis by Western blot, or extracted for perlecan isolation and subsequent characterization of glycosaminoglycan size and disaccharide composition. 35SO4 incorporation into perlecan was three-fourfold higher in the proliferating/hypertrophic zone than the resting zone. Western blot showed perlecan content was greatest in the lower and upper proliferating zones and that a perlecan fragment lacking portions of the N- and C-terminal domains containing heparan sulfate was also present in all zones. Purified perlecan from the hypertrophic/lower proliferative zone had larger chondroitin sulfate chains and a different composition of CS and HS disaccharides than the perlecan isolated from the resting zone. These results indicate perlecan deposition is increased and is turned over during proliferation to be replaced by a perlecan with a different sulfation pattern. PMID- 16705695 TI - Endochondral growth in growth plates of three species at two anatomical locations modulated by mechanical compression and tension. AB - Sustained mechanical loading alters longitudinal growth of bones, and this growth sensitivity to load has been implicated in progression of skeletal deformities during growth. The objective of this study was to quantify the relationship between altered growth and different magnitudes of sustained altered stress in a diverse set of nonhuman growth plates. The sensitivity of endochondral growth to differing magnitudes of sustained compression or distraction stress was measured in growth plates of three species of immature animals (rats, rabbits, calves) at two anatomical locations (caudal vertebra and proximal tibia) with two different ages of rats and rabbits. An external loading apparatus was applied for 8 days, and growth was measured as the distance between fluorescent markers administered 24 and 48 h prior to euthanasia. An apparently linear relationship between stress and percentage growth modulation (percent difference between loaded and control growth plates) was found, with distraction accelerating growth and compression slowing growth. The growth-rate sensitivity to stress was between 9.2 and 23.9% per 0.1 MPa for different growth plates and averaged 17.1% per 0.1 MPa. The growth-rate sensitivity to stress differed between vertebrae and the proximal tibia (15 and 18.6% per 0.1 MPa, respectively). The range of control growth rates of different growth plates was large (30 microns/day for rat vertebrae to 366 microns/day for rabbit proximal tibia). The relatively small differences in growth-rate sensitivity to stress for a diverse set of growth plates suggest that these results might be generalized to other growth plates, including human. These data may be applicable to planning the management of progressive deformities in patients having residual growth. PMID- 16705696 TI - Inhibitory effects of a new bisphosphonate, minodronate, on proliferation and invasion of a variety of malignant bone tumor cells. AB - Little is known about the biological effects of bisphosphonates on primary malignant bone tumors. The purpose of this study was to investigate the antitumor effects of newly developed minodronate (MIN) on a variety of human malignant bone tumors. We examined the effects of MIN and clinically relevant incadronate (INC) on the proliferation, apoptosis, and cell cycle of two osteosarcoma (Saos-2, MG 63), two chondrosarcoma (SW1353, OUMS27), and two Ewing's sarcoma (RD-ES, SK-ES 1) cell lines. Furthermore, we investigated the anti-invasion effects of MIN on sarcoma cells and the effects of MIN on tumor growth in nude mice. MIN inhibited the viability of all six cell lines in a dose-dependent manner with IC50 values of 2.7 to 5.0 microM, which were significantly lower than those of INC. Importantly, both bisphosphonates affected the viability of normal bone marrow stromal cells much less than sarcoma cells. Both bisphosphonates induced cell cycle perturbation in all sarcoma cells tested and apoptosis in Saos-2 and SW1353 cells, although they failed to induce apoptosis in RD-ES and SK-ES-1 cells. MIN significantly suppressed invasion, even at a low concentration of 1 microM (p < 0.01). Daily injection of 5 microg of MIN inhibited the growth of SK-ES-1 xenograft sarcoma in nude mice without loss of body weight. These findings suggest that MIN may have a beneficial adjuvant role in the treatment of patients with malignant bone tumors. PMID- 16705697 TI - Apoptosis induced by chemotherapeutic agents involves c-Jun N-terminal kinase activation in sarcoma cell lines. AB - Molecular mechanisms underlying chemotherapeutic agent-induced apoptosis in sarcoma cells are not well known. Induction of apoptosis is regulated by several components including mitogen-activated protein kinases (MAPKs) comprising ERK, p38MAPKs, and c-Jun N-terminal kinase (JNK). In the present study, we examined whether activation of JNK is induced by the chemotherapeutic agents cis diaminedichloroplatinum (cisplatin, CDDP) or doxorubicin (DXR), and whether the ectopic expression of constitutively active (MKK7-JNK1) or dominant-negative form of JNK (dnJNK) influenced apoptosis in response to the CDDP or DXR in sarcoma cell lines MG-63 and SaOS-2. The CDDP or DXR induced JNK activation in the both cell lines, as assessed by Western blotting using phosphospecific antibodies. A transient expression of the activated form of JNK sensitized the MG-63 and SaOS-2 cells to the drug-induced apoptosis, while dnJNK1 reduced the proportion of apoptotic cell death. Apoptosis was determined by flow cytometry using annexin-V Cy5. Collectively, our results indicate that JNK activation is involved in apoptotic cell death in sarcoma cell lines following stimulation with CDDP or DXR. PMID- 16705698 TI - Doxorubicin induces cell senescence preferentially over apoptosis in the FU-SY-1 synovial sarcoma cell line. AB - Surgical resection coupled with adjuvant radiotherapy and/or doxorubicin based chemotherapy are the mainstays of synovial sarcoma (SS) treatment. Although effective as a SS adjuvant, the proposed mechanism of action of doxorubicin remains controversial. Current opinion supports DNA damage-induced apoptosis. This in vitro study used cDNA gene expression profiling to investigate whether apoptosis, alone or in combination with cell senescence, is induced by doxorubicin in SS cells. Cell cultures of the FU-SY-1 SS, the pleomorphic SW982 sarcoma, and a primary dermal fibroblast (NHDF), were exposed to 500 nM doxorubicin, and then processed for cDNA microarray analysis. The one class response option of SAM (Significance Analysis of Microarrays) was used to test for significant overexpression of 15 apoptosis-related genes and nine senescence related genes. Drug-induced cell senescence was quantified by measuring beta galactosidase activity. None of 15 apoptosis-related genes and only two of nine senescence-related genes were identified by SAM as significantly overexpressed in doxorubicin-treated cultures. Drug-induced senescence as reflected by beta galactosidase activity was significantly increased (p < 0.05) only in FU-SY-1 SS cultures. Apoptosis does not appear to be a major determinant of doxorubicin induced mortality in FU-SY-1 SS or NHDF cultures, but may impact SW982 cells via the overexpression of BAX relative to Bcl-2. Doxorubicin-induced cell senescence was prominent in FU-SY-1 SS cultures, but negligible in SW982 and NHDF cultures. Likely, both apoptosis and cell senescence contribute to doxorubicin-induced cell death in this synovial sarcoma cell line. PMID- 16705699 TI - The cypriot and Iranian National Mutation Frequency Databases. AB - The National Mutation Frequency Databases are continuously updated mutation depositories, which contain extensive information over the described genetic heterogeneity of an ethnic group or population. Here, we report the construction of the Cypriot (http://www.goldenhelix.org/cypriot) and Iranian National Mutation Frequency Databases (http://www.goldenhelix.org/iranian), both derived from an academic effort to provide high quality and up-to-date information on the underlying genetic heterogeneity of inherited disorders in the Cypriot and Iranian populations, respectively. Both databases have been built and maintained online using ETHNOS platform, a specialized software, which provides the means for national mutation database construction and curation. Each database contains brief summaries of the various genetic disorders studied for each population, and an easy-to-use query interface provides, both to specialist as well as to non specialist users (i.e. patients and their families), instant access to the list and frequencies of the different mutations responsible for the inherited disorders in these populations. Furthermore, numerous links to the respective Online Mendelian Inheritance in Man (OMIM) entries and, when available, to the locus-specific databases fruitfully integrate the databases content into a single Web site. Both databases can serve as valuable online tools for molecular genetic testing of inherited disorders in these populations and could potentially motivate further investigations of yet unknown genetic diseases in the Cypriot and Iranian populations. PMID- 16705700 TI - Release of nitric oxide, but not prostaglandin E2, by bone cells depends on fluid flow frequency. AB - Loading frequency is an important parameter for the stimulation of bone formation in vivo. It is still unclear how the information of external loading characteristics is conveyed to osteoblasts and osteoclasts. Osteocytes are thought to detect mechanical loads by sensing fluid flow through the lacuno canalicular network within bone and to translate this information into chemical signals. The signaling molecules nitric oxide (NO) and prostaglandin E2 (PGE2) are known to play important roles in the adaptive response of bone to mechanical loads. We have investigated the effects of fluid flow frequency on the production of PGE2 and NO in bone cells in vitro. Pulsatile fluid flow with different frequencies stimulated the release of NO by MC3T3-E1 osteoblasts in a dose dependent manner. In contrast, PGE2 production was enhanced consistently by all fluid flow regimes, independent of flow frequency. This implies that the NO response may play a role in mediating the differential effects of the various loading patterns on bone. PMID- 16705701 TI - Moderate soft tissue trauma delays new bone formation only in the early phase of fracture healing. AB - The aim of this study was to investigate the effect of a moderate soft tissue trauma to the course of fracture healing in a standardized animal model. Thirty eight Wistar rats were randomly divided into a fracture group (F, n = 19) and a group with a fracture and a soft tissue trauma (F + STT, n = 19). The fracture and the soft tissue trauma were created using an impact device with a standardized energy. All fractures were stabilized by two Kirschner wires. Three rats were measured for blood flow and sacrificed at days 1, 3, 7, and 14, and seven rats at day 28, from both groups. A three-point bending test was performed on the healed tibia after 28 days. During the first 24 h there was a reduction in blood flow, which was more pronounced in the F + STT group than in the F group. From histological sections, the shape of the callus formation, as well as the tissue distribution of newly formed bone, fibrous cartilage and fibrous connective tissue were determined. Distinctly more periosteal new bone formed and a larger callus formed at days 3 and 7 in group F compared to group F + STT. However, by days 14 and 28, the ossification and overall callus size no longer showed differences between the two groups. A fast recovery of blood flow and callus formation took place in the F + STT group, which led to similar histological and biomechanical results in fracture healing observed after 28 days between the two groups. PMID- 16705702 TI - Variability in tendon and knee joint biomechanics among inbred mouse strains. AB - Hereditary factors are thought to be responsible for impaired tendon function and joint laxity. The present study investigated the genotypic variability of knee laxity and stiffness and tendon mechanical and geometric properties among 16-week old female A/J, C57BL/6J (B6), and C3H/HeJ (C3H) inbred mice. In one group of mice, knee mechanics were quantified using a custom loading apparatus enabling translation of the tibia against a stationary femur. In a second group, flexor digitorum longus and Achilles tendons from the left hind limb underwent biomechanical testing, while those of the contralateral limb were analyzed histologically for determination of cross-sectional area. Our results demonstrate that tendon and joint mechanics varied significantly among the inbred mouse strains, indicating that biomechanical properties are genetically determined. A/J mouse knees exhibited greater laxity (p < 0.001) and lower stiffness (p < 0.001) compared to those of the B6 and C3H mice. The genotypic differences in whole joint properties were similar to those of the tendons' structural biomechanical traits. Although body mass did not differ (p > 0.2) among the three strains, significant genotypic differences were found at the whole tendon, material quality, and morphological levels of the tissue hierarchy. Furthermore, genetic regulation of tendon mechanical properties varied with anatomic site. Patterns of genotypic differences in tendon size were not consistent with those of biomechanical properties, suggesting that unique combinations of structural and compositional factors contribute to tendon growth, adaptation, and development. Therefore, the three inbred strains constitute a useful experimental model to elucidate genetic control of structure-function relationships in normal and healing tendons and ligaments. PMID- 16705703 TI - An investigation into the effect of number of trials during proprioceptive testing in patients with patellofemoral pain syndrome. AB - The purpose of this study was to investigate the effect of increasing the number of test trials in the assessment of knee joint position sense, in a group of patellofemoral patients, and second, to investigate the effects of joint angle and type of test. Thirty-two patients with a diagnosis of patellofemoral pain syndrome were admitted to this study. Knee joint position sense was measured by performing an ipsilateral matching activity at target angles of 20 degrees and 60 degrees using two techniques; Passive Angle Reproduction (PAR), and Active Angle Reproduction (AAR). The results show that five repetitions are required for AAR and six for PAR. A two-way repeated-measures ANOVA showed that there was no significant difference (p = 0.559) in the accuracy of angle reproduction for the two angles (60 degrees and 20 degrees ); however, there was a significant difference (p = 0.001) between the two types of test (AAR and PAR). The results of this study have implications for clinicians and researchers who examine knee joint proprioception as part of functional assessment in a variety of knee joint pathologies including PFPS. A "quick" one-off assessment of knee joint position sense may give erroneous data; subjects need to have several trials before a true picture of their proprioceptive status emerges. Type of testing performed, active or passive, is also important, although the angle of testing may not be relevant. PMID- 16705704 TI - An Er:YAG laser endoscopic fiber delivery system for lithotripsy of salivary stones. AB - BACKGROUND AND OBJECTIVES: Endoscopic applications of Erbium:YAG lasers are still very limited due to lack of appropriate fiber delivery capabilities. Recent reports on potential advantages of this laser for lithotripsy of ureteral stones prompted us to develop an Er:YAG fiber delivery system for endoscopic lithotripsy of salivary stones. We report on the development of this system and its clinical use on 17 patients. STUDY DESIGN/MATERIALS AND METHODS: Ho:YAG and Er:YAG laser fragmentation performances were initially compared. Optimal laser parameters for lithotripsy of salivary stones were then established ex vivo using a commercial dental Er:YAG laser (Lumenis Opusdent 20). Metal hollow waveguides optimized for Er:YAG laser transmission were end sealed with a polished sapphire rod of 0.63 mm diameter and designed to adapt to the Opusdent laser and to a Storz sialoendoscope. The system was tested ex vivo for durability and clinical compatibility at input energies up to 700 mJ, 10-20 Hz. Following Helsinki approval the system was clinically tested on 17 patients with sialolithiasis. RESULTS: Lithotripsy threshold was around 80 mJ/pulse (26 J/cm2) while efficient fragmentation, with microscopic fragments, was observed at an output energy range of 150-300 mJ/pulse. At 10 Hz, fragmentation rates of about 1.8 mm3/second were achieved enabling lithotripsy of a 6 mm stone in about 2 minutes. Front surface damage to the sapphire rod occurred but did not contribute to significant loss in fragmentation efficiency. Of the 21 stones treated clinically, 5 were fully fragmented, 7 were prepared for extraction by mini forceps, and 9 were released from surrounding soft tissues for subsequent removal. Fifteen of the 18 treated glands returned to normal function without any symptoms. CONCLUSIONS: The Er:YAG endoscopic delivery system described is a clinically viable and cost-effective device for a range of hard and soft tissue wet field applications accessible through rigid or semi-rigid endoscopes. Further improvements in the waveguide may allow access also through fully flexible endoscopes. PMID- 16705705 TI - Nerve endings of the wrist joint: a preliminary report of the dorsal radiocarpal ligament. AB - As part of an investigation of the articular nerve ending populations in the wrist joint capsule associated with the anterior and posterior interosseous nerves, this study addresses the nerve ending population in the dorsal radiocarpal ligament. The ligaments were harvested from four wrists of two fresh cadavers within 12 h of death. Tissues were fixed, cryostat sectioned, and processed for fluorescence immunohistochemistry using antibody to protein gene product 9.5 (PGP 9.5), a general or pan neuronal marker, and a secondary antibody conjugated to a fluorescent tag (Alexa Fluor 488). The sections were evaluated with a confocal laser microscope and an image analyzer. Labeled nerve endings were mapped, measured, and categorized. Type I (Ruffini-like ending), Type III (Golgi-like tendon organ) and Type IV (noncorpuscular) nerve endings could be identified in all four DRC ligaments, with Types I and IV dominating. These receptors were distributed primarily over the superficial two thirds of the ligament (>80%), and near the bony attachments (>70%). The dorsal radiocarpal ligament has a rich sensory innervation from the posterior interosseous nerve terminating in nerve endings located in the superficial two-thirds of the ligaments, primarily near bony attachment sites. PMID- 16705706 TI - Longitudinal quantitative evaluation of lesion size change in femoral head osteonecrosis using three-dimensional magnetic resonance imaging and image registration. AB - It remains controversial whether some lesions of femoral head osteonecrosis regress during the natural course of the disease. With image registration, accurately matched image sets of the same subject can be acquired at different times. We applied image registration to evaluate lesion size change and assessed accuracy and usefulness compared to volume measurements and a conventional method. We also investigated whether lesions regress with this technique and with volume measurements. Baseline and 1 year minimum follow-up scans were conducted on 25 patients (31 hips) without radiological evidence of collapse. A three dimensional (3D) spoiled gradient recalled echo sequence was used in the coronal direction (slice thickness = 2 mm; slice pitch = 1 mm). Size change was evaluated on all contiguous pairs of matched images after image registration. As a conventional method, coronal images (slice thickness = 5 mm) were reconstructed, and size change was evaluated on the five representative coronal slices. Evaluation with the conventional method identified eight lesions with apparent reduction; assessments using image registration and volume measurements identified three lesions, all within a year of initial steroid treatment and remaining at ARCO stage I at follow up. Evaluation of lesion size change using image registration was comparable to volume measurements. Inaccurate estimation of lesion size change due to mismatching of slice planes can be excluded. We demonstrated that some early lesions detected less than a year after initial steroid treatment can show size reduction with image registration as well as with volume measurements. PMID- 16705708 TI - Effect of oral diacerein (DAR) in an experimental hip chondrolysis model. AB - We aimed to reproduce the articular cartilage structural changes in a joint exposed to a metallic implant as in the adolescent pinned hip with persistent joint penetration and secondly, to test the effect of an interleukin inhibitor, diacerein (DAR) in the ensuing articular cartilage lesion. Twenty immature beagles were submitted to a surgical K-wire implantation in the hip with the material left in the joint space for 6 months. Twelve animals were sacrificed for histological and biochemical tests. Eight animals were sacrificed at 10 months (half of them treated with DAR) and analyzed by scanning electron microscopy (SEM) and biochemistry of the articular cartilage. Preoperative and monthly C3 and C4 complement and immunoglobulins serum levels were determined. The histological and the electrophoretic profile changes were significative at 6 months. At 10 months the migration profile (CaCl2) recovered to normal levels in the operated hip and the SEM scores for the acetabulum were similar to the non operated control hip after treatment. The serum level of IgA was elevated at the 4th and 6th month postoperatively. The persistence of a metallic implant resulted in degenerative changes parallel to that described for hip chondrolysis as a complication of in-situ pinning; and the cartilage lesion improved with DAR treatment. PMID- 16705709 TI - Influence of ovarian hyperstimulation and ovulation induction on the cytoskeletal dynamics and developmental competence of oocytes. AB - This study was undertaken to determine the effects of gonadotrophin on cytoskeletal dynamics and embryo development and its role in improving the retrieval of developmentally competent oocytes. Female golden hamsters were injected with human chorionic gonadotrophin (hCG; 5-, 7.5- or 15-IU) on the day 4 of estrus, pregnant mare serum gonadotrophin (PMSG; 5-, 7.5- or 15-IU) on the day 1 of estrus, or 15-IU hCG at 56 hr post-15-IU PMSG injection in any cycle except estrus. Increasing the hCG dose decreased not only retrieval rate of 2-cell embryo but development to blastocyst after subsequent in vitro culture. Whereas, although increasing the PMSG dose induced increasing the number of 2-cell embryo and blastocyst, 15-IU PMSG injection caused retardation of development to blastocyst. No 2-cell embryos were retrieved by injecting both PMSG and hCG. The injections of 15-IU hCG and 7.5- or 15-IU PMSG inhibited the proliferation of trophectodermal and inner cell mass cells, respectively. Gonadotrophin injection didn't influence microtubular spindle formation, but 5- or 15-IU hCG, 15-IU PMSG, or PMSG and hCG injections induced aberrant cortical granule (CG) and microfilament distribution. After 15-IU hCG or PMSG and hCG injections, fewer oocytes had enriched cortical actin domains, and the expression of alpha-, beta- and gamma-actin genes was greatly increased. In conclusion, a high dose of gonadotrophins alters the microfilament and CG distribution, which in turn reduces the developmental competence of oocytes. Injecting a reduced dose of PMSG to initiate ovarian hyperstimulation without triggering ovulation contributes to the efficient retrieval of developmentally competent oocytes. PMID- 16705710 TI - Global gene profiling reveals a downregulation of BMP gene expression in experimental atrophic nonunions compared to standard healing fractures. AB - Nonunion is a challenging problem that may occur following certain bone fractures. However, there has been little investigation of the molecular basis of nonunions. Bone morphogenetic proteins (BMPs) play a significant role in osteogenesis. However, little is known about the expression patterns of BMPs in abnormal bone healing that results in nonunion formation. These facts prompted us to investigate and compare the gene expression patterns of BMPs and their antagonists in standard healing fractures and nonunions using rat experimental models. Standard closed healing fractures and experimental atrophic nonunions produced by periosteal cauterization at the fracture site were created in rat femurs. At postfracture days 3, 7, 10, 14, 21, and 28, total RNA was extracted from the callus of standard healing fracture and fibrous tissue of nonunion (n=4 per each time point and each group). Gene expression of BMPs, BMP antagonists, and other regulatory molecules were studied by methods including Genechip microarray and real-time quantitative RT-PCR. Gene expression of BMP-2, 3, 3B, 4, 6, 7, GDF-5, 7, and BMP antagonists noggin, drm, screlostin, and BAMBI were significantly lower in nonunions compared to standard healing fractures at several time points. Downregulation in expression of osteogenic BMPs may account for the nonunions of fracture. The balance between BMPs and their endogenous antagonists is critical for optimal fracture healing. PMID- 16705711 TI - Novel sequence variants in dysferlin-deficient muscular dystrophy leading to mRNA decay and possible C2-domain misfolding. AB - Mutations in the gene encoding dysferlin (DYSF) cause the allelic autosomal recessive disorders limb girdle muscular dystrophy 2B and Miyoshi myopathy. It encompasses 55 exons spanning 150 kb of genomic DNA. Dysferlin is involved in membrane repair in skeletal muscle. We identified three families with novel sequence variants in DYSF. All affected family members showed limb girdle weakness and had reduced or absent dysferlin protein on immunohistochemistry. All exons of DYSF were screened by genomic sequencing. Five novel variants in DYSF were found: two missense mutations (c.895G>A and c.4022T>C), one 5' donor splice site variant (c.855+1delG), one nonsense mutation (c.1448C>A), and a variant in the 3'UTR of DYSF (c.*107T>A). All alterations were confirmed by restriction enzyme analysis and not found in 400 control alleles. Nonsense mediated RNA decay or changes in the three-dimensional protein structure resulting in intracellular dysferlin aggregates and finally the lack of dysferlin protein were identified as consequences of the novel DYSF variants. PMID- 16705712 TI - Molecular bases of antithrombin deficiency: twenty-two novel mutations in the antithrombin gene. AB - Antithrombin (AT) is a major physiological inhibitor of hemostasis. We report 22 novel antithrombin gene (SERPINC1) mutations associated with antithrombin deficiency in 17 French and five German families. They were all present at the heterozygous state. Nine missense mutations accounted for type I deficiency, defined by equally low antithrombin activity and antigen level. Most of them (7/9) affected highly conserved serpin residues and were associated with venous thrombosis occurring at a young age (before age 32). One splice site, one nonsense mutation, three small deletions and one insertion were also identified as a cause for type I antithrombin deficiency. Seven other missense mutations were identified in type II or unclassified AT deficiency; g.5270C>T (p.T147I, T115I) and g.5281A>T (p.I151F, I119F) change residues in the heparin binding region, g.13267C>G (p.P439A, P407A) and g.13271T>C (p.F440S, F408S) affect amino acids in the pleiotropic region, g.2372G>A (p.G25D, G-8D) changes a signal peptide amino acid, g.2456G>C (p.C53S, C21S) affects one of the three disulfide bonds of the protein, and g.7585A>T (p.M347K, M315K) changes a nonconserved residue on strand 2C. PMID- 16705713 TI - Hepatic and neuromuscular forms of glycogenosis type III: nine mutations in AGL. AB - Glycogenosis type III (Cori disease) is an autosomal recessive disorder caused by the deficiency of the glycogen debranching enzyme, encoded by the AGL gene, and existing in six isoforms alternately spliced in a tissue-specific way. Generally, disease onset occurs early on starting from the first year of life, with hepatomegaly, hypoglycemia, hyperlipidemia, increased CK levels, and, in some cases, short stature and slight mental retardation. Frequently, hepatomegaly tends to resolve spontaneously and inexplicably during childhood, when myopathy, often associated with cardiomyopathy, arises. This disease is known to lack almost invariably clear links between the genotype and clinical phenotype. We describe nine new mutations in Italian patients: four nonsense (p.Arg285X, p.Lys422X, p.Arg910X, p.Arg977X), three frameshift (c.442delA, c.753_756delGACA, c.3963delG), and two missense (p.Ala1120Pro, p.Arg524His). Particularly, the nonsense p.Arg285X is linked to an exonic splicing enhancer and it was found to produce two species of transcripts at the same time. Moreover, we discuss a subgroup of subjects carrying c.2681+1G>A, which has proven to be the most frequent mutation among our patients. The previously described c.664+3A>G was also detected in two patients, both homozygous. The present work is yet another confirmation that the individual genetic background plays a pivotal role in influencing the phenotypes, as occurs in other metabolic diseases. PMID- 16705714 TI - Effect of knee joint angle on motor unit synchronization. AB - Activity of the vasti has been argued to vary through knee range of movement due to changes in passive support of the patellofemoral joint and the relative contribution of these muscles to knee extension. Efficient function of the knee is dependent on optimal control of the patellofemoral joint, largely through coordinated activity of the medial and lateral quadriceps. Motor unit synchronization may provide a mechanism to coordinate the activity of vastus medialis (VMO) and vastus lateralis (VL), and may be more critical in positions of reduced passive support for the patellofemoral joint (i.e., full extension). Therefore, the aim of this study was to determine whether the degree of motor unit synchronization between the vasti muscles is dependent on joint angle. Electromyographic (EMG) recordings of single motor unit action potentials (MUAPs) were made from VMO and multiunit recordings from VL during isometric contractions of the quadriceps at 0 degrees, 30 degrees, and 60 degrees of knee flexion. The degree of synchronization between motor unit firing was evaluated by identification of peaks in the rectified EMG averages of VL, triggered from MUAPs in VMO. The proportion of cases in which there was a significant peak in the triggered averages was calculated. There was no significant difference in the degree of synchronization between the vasti at different knee angles (p=0.57). These data suggest that this basic coordinative mechanism between the vasti muscles is controlled consistently throughout knee range of motion, and is not augmented at specific angles where the requirement for dynamic control of stability is increased. PMID- 16705715 TI - Retrieval, experimental, and computational assessment of the performance of total knee replacements. AB - Wear mechanisms in polyethylene components for total knee replacements are inherently mechanical; the local stresses or strains exceed some material limit. Retrieval analysis and knee simulators have provided the means to quantify the damage observed in vivo or in vitro. These results have been circumstantially linked to the material stresses obtained from computational simulations using finite element analysis, knee simulator tests, and computational simulations of two condylar knee designs. We hypothesize that if an equivalent loading environment is produced in the computational simulation, we can correlate the distribution of computed stresses with observed damage of simulator specimens and further relate design differences to in vivo performance from retrieval analyses. The finite element model agreed with the knee simulator kinematics and kinetics within 2-13%, and composite FEA contact areas matched 66-90% of the damage areas due to burnishing on the simulator specimens. Burnishing was the primary mode of damage for both the simulator and retrieval specimens corresponding with the relatively low magnitudes of contact stress observed. Both the computational and experimental techniques underpredicted the amount determined from retrieval analysis, but the differences between the two designs were consistent for all three methods. Combining these techniques strengthens the applicability of the computational simulation while highlighting the complementary approach of these methods for preclinical testing and assessing the link between material state and damage. PMID- 16705716 TI - Effect of compliant flooring on impact force during falls on the hip. AB - Compliant flooring represents a promising but understudied strategy for reducing impact force and hip fracture risk due to falls in high-risk environments such as nursing homes, hospitals, gymnasiums, and senior centers. We conducted "pelvis release experiments" with young women (n=15) to determine whether floor stiffness influences peak hip impact force during safe, low-height falls. During the trials, we used a pelvic sling and electromagnet to lift and instantly release the participant from a height of 5 cm above a force plate, which measured the force applied to the hip region during impact. Trials were conducted for rigid floor conditions and with layers of ethylene vinyl acetate foam rubber overlying the floor that we regarded as firm (1.5-cm thick; stiffness=263 kN/m), semifirm (4.5-cm thick; stiffness=95 kN/m), semisoft (7.5-cm thick; stiffness=67 kN/m), and soft (10.5-cm thick; stiffness=59 kN/m). When compared to the rigid condition, peak hip impact force averaged 8% lower in the firm condition and 15% lower in the semifirm condition. Peak forces were not significantly different between the semifirm, semisoft, and soft floor conditions, indicating that a 4.5 cm-thick foam mat provides nearly the same force attenuation as a 10.5 cm-thick mat. These results support the need for laboratory experiments to measure the effect of floor stiffness on postural stability and for clinical trials to determine the effect of compliant flooring on hip fracture incidence in high-risk environments. PMID- 16705717 TI - Inhibition of IKK activation, through sequestering NEMO, blocks PMMA-induced osteoclastogenesis and calvarial inflammatory osteolysis. AB - Osteoclasts, the primary bone-resorbing cells, play a crucial role in periprosthetic bone loss in response to implant-derived wear debris. Differentiation and activation of osteoclasts at the implant-bone interface are fueled by elevated levels of locally secreted inflammatory cytokines that heighten the osteolytic response. Among these cytokines are members of the TNF superfamily, including TNF and RANKL, which primarily act through activation of the transcription factor NF-kappaB. Activation of NF-kappaB is required for osteoclast formation, and its inhibition hampers osteoclastogenesis and bone loss. Activation of NF-kappaB is permitted following its dissociation from the inhibitory protein IkappaBalpha, an event subsequent to phosphorylation of the latter protein by the upstream IkappaBalpha kinase (IKK) complex. Our recent findings show that attenuating IKK complex assembly, by using a short peptide termed NEMO-binding domain (NBD) peptide, that blocks binding of IKK2 and IKK1 to IKKgamma/NEMO, inhibits NF-kappaB activation, and arrests RANKL-induced osteoclastogenesis. In this study, we examined if NBD is capable of blocking inflammatory osteolysis by PMMA particles. Our findings indicate that NBD peptide inhibits PMMA-induced IKK2 and NF-kappaB activation. More importantly, this peptide potently arrests PMMA-stimulated osteoclastogenesis and alleviates PMMA induced inflammatory and osteolytic responses in mice. Thus, NBD peptide is considered as a promising modality to regulate inflammatory osteolysis. PMID- 16705718 TI - Interactions between microstructural and geometrical adaptation in human cortical bone. AB - With aging and in disease, the changes in bone microstructure and geometry influence the mechanical properties of cortical bone, however, the level of interaction between the two is not known. Here, we investigate the interaction between the changes in microstructural and geometrical properties of the aging male tibia in proximal and distal middiaphysis. The microstructural measurements include variables related to the size and density of osteons and intracortical porosity. The macroscopic geometrical properties include variables related to bone surfaces (periosteal and endosteal) and cross section (area, moment of inertia). Site-specific correlations were found between the microstructural and geometrical properties along the bone length and at different bone surfaces. In contrast to the proximal middiaphysis of male tibia, where no correlation existed, significant (p<0.05) correlations were found in the distal middiaphysis of tibia. The changes in parameters partially related to bone formation in the cortex, including the osteonal area, showed positive correlations with an increase in the periosteal diameter. Similarly, parameters related to bone resorption and/or failed formation in the cortex, including porosity and pore size, showed significant correlations with cortical thinning. These findings support the concept that, with aging, anabolic and catabolic responses in the human tibia at microstructural and macrostructural levels are spatially related and site specific. PMID- 16705719 TI - Alendronate prevents femoral periprosthetic bone loss following total hip arthroplasty: prospective randomized double-blind study. AB - Following total hip arthroplasty (THA), femoral periprosthetic bone undergoes a remodeling process that results in bone loss in its proximal regions that may compromise the long-term outcome of THA. Periprosthetic bone loss mainly occurs during the first postoperative months. The question is whether a postoperative treatment with alendronate is effective in reducing periprosthetic bone loss and which doses and duration of treatment are required. In a 12-month prospective, randomized double-blind study, 51 patients undergoing cementless THA were treated postoperatively either with a daily dose of 20 mg alendronate for 2 months and 10 mg for 2 months thereafter (group I), with 20 mg of alendronate for 2 months and 10 mg for 4 months thereafter (group II), or treated with placebo (group III). Proximal femoral bone mineral density (BMD) was measured with dual-energy X-ray absorptiometry (DEXA) and serum biochemical markers of bone turnover bone specific alkaline phosphatase, osteocalcin, and C-terminal telopeptides (CTX-I) were assayed. Six months of alendronate treatment significantly reduced (p<0.001) bone loss in proximal medial region (-10%) compared with placebo (-26%). All biochemical markers of bone turnover were suppressed by alendronate. These data suggest that alendronate administered for the first 6 postoperative months following THA was effective in preventing early periprosthetic bone loss. PMID- 16705720 TI - Upregulated expression of inducible nitric oxide synthase plays a key role in early apoptosis after anterior cruciate ligament injury. AB - The reason that the anterior cruciate ligament (ACL) has a very poor healing potential after injury is not well understood. In this study, we investigated the role of nitric oxide (NO) in the apoptotic cell death of ACL cells using a rabbit model and in vitro cell culture. The apoptosis of ACL cells in vivo was analyzed by TUNEL assay and electron microscopy. NO synthase (NOS) expression was observed by immunohistochemical analysis. ACL cells were cultured and the susceptibility to NO-induced apoptosis was tested. Inducible NOS (iNOS) expression after treatment with cytokines was examined by immunohistochemical and RT-PCR analyses. Mitogen-activated protein kinase (MAPK) inhibitors were used for the analysis of downstream signals. A significant number of apoptotic cells were observed on days 1 to 3 after injury; the apoptotic rate returned to the control level by day 7. Upregulation of iNOS in the ACL remnant was observed at day 1. Intraarticular injection of NOS inhibitor suppressed the apoptotic rate. Isolated ACL cells showed much higher susceptibility to NO-induced apoptosis than did medial collateral ligament cells. IL-1beta stimulated ACL cells to upregulate iNOS mRNA and increase NO production. p38 MAPK inhibitor decreased NO-induced apoptosis. Rapid iNOS induction after injury contributes to the high apoptotic rate of ACL cells, and this may partly account for the poor healing capacity of this ligament. iNOS and NO production is suggested to be stimulated by IL-1beta, and NO activates the p38 MAPK pathway and triggers an apoptotic signal in ACL cells. PMID- 16705721 TI - The wait. PMID- 16705722 TI - Preoperative chemoradiation in rectal cancer: Retrospective comparison between capecitabine and continuous infusion of 5-fluorouracil. AB - BACKGROUND: We compared the efficacy and toxicity of oral capecitabine and continuous infusion of 5-fluorouracil (5-FU) in the preoperative chemoradiation treatment of patients with rectal cancer. PATIENTS AND METHODS: The files of 89 patients with rectal cancer, 43 treated preoperatively with oral capecitabine and 46 with intravenous 5-FU, were reviewed, and the outcome of the groups was compared. RESULTS: There was no statistically significant difference in the complete pathological response rate between the capecitabine and the 5-FU groups (30% vs. 17%, P = 0.15). The downstaging rate was higher in the capecitabine group (77% vs. 50%, P = 0.009). Toxicity was mild in both groups. The rate of Grade 3 gastrointestinal toxicity was similar in the two groups (diarrhea 2% vs. 4%, proctitis 5% vs. 7%), except for one patient in the 5-FU group (2%) who developed a rectovaginal fistula. In the capecitabine group, one patient (2%) had Grade 3 hand-foot syndrome, and another had an acute myocardial infarction. In the 5-FU group, two patients (4%) had Grade 3 hematological toxicity, and three (6%) had complications from Port-a-Cath insertion. CONCLUSION: Preoperative chemoradiation with oral capecitabine appears to be safe and well tolerated, and at least as good as continuous 5-FU. PMID- 16705723 TI - Can axillary dissection be avoided in patients with sentinel lymph node metastasis? AB - BACKGROUND: Who should undergo a completion dissection following identification of a +sentinel lymph node (SLN) is controversial. METHODS: The records of 1,133 patients who underwent SLN mapping were reviewed. The association between patient, tumor, and treatment characteristics and the presence of +SLNs and +nonSLNs was analyzed using two-way tables of frequency counts and Pearson chi2 test. Possible predictors of +SLNs and +nonSLNs were analyzed using simple and multiple logistic regression. RESULTS: One thousand one hundred forty-eight SLN procedures were performed. 367 procedures (32%) yielded +SLNs. For patients with a +SLN, on multiple logistic regression analysis LVSI, increasing numbers of +SLNs, decreasing numbers of negative SLNs, and increasing size of the largest SLN metastasis were statistically significantly associated with increased likelihood of nonSLN involvement. No subgroup was identified that did not have a significant rate of nonSLN involvement on completion axillary dissection, except those who had a large number of negative SLNs (> or =3) and small size of the largest SLN metastasis (<10 mm). CONCLUSIONS: A definitive answer to the question of who needs a completion axillary dissection awaits the results of ongoing trials. In the interim, our data does not support eliminating dissection for any subgroup of patients with +SLNs. PMID- 16705724 TI - Does sentinel lymph node biopsy in cutaneous head and neck melanoma alter disease outcome? AB - BACKGROUND AND OBJECTIVES: In the head and neck region, value, reliability, and safety of sentinel lymph node biopsy (SLNB) have not yet been determined conclusively. The aim of study was to assess impact of SLNB on disease outcome in cutaneous head and neck melanoma. METHODS: Thirty-six patients with a clinically node-negative head and neck melanoma, > or =1.0 mm Breslow thickness, participated in a prospective study from 1995 to 2005. Sentinel lymph node (SLN) tumor-positive patients underwent completion lymphadenectomy. SLN tumor-negative patients underwent clinical monitoring. Median follow-up was 54 (range 10-114) months. Recurrence-free and overall survival curves were constructed by Kaplan Meier. RESULTS: SLNs could be identified in 33 patients (92%). In 7 patients (21%) the SLN was tumor-positive. In 1 patient (13%) the SLNB was false-negative. In 17 patients (47%) SLNs could be identified in the parotid region (success rate parotid region 100%). This study showed no significant difference in recurrence free and overall survival between patients with tumor-positive and tumor-negative SLN. CONCLUSIONS: The safety and accuracy of SLNB in the neck and parotid nodal basins were similar to those in non-head and neck sites. However, the technique is technically demanding in this region. In this small series SLNB did not alter disease outcome. PMID- 16705725 TI - Quality of life in newly diagnosed cancer patients waiting for surgery is seriously impaired. AB - BACKGROUND AND OBJECTIVES: Newly diagnosed cancer patients waiting for initial surgery experience a stressful time. Study objectives were (1) to examine the quality of life (QL) of these patients by comparing QL (a) between the four diagnostic groups included in this study, (b) to the QL of the general population, (2) to determine the factors that contribute most to patients' overall QL. METHODS: One hundred ninety six patients with lung, periampullary, oesophageal and cervical cancer completed questionnaires on generic QL (SF-36), overall QL, cancer-site specific symptoms (EORTC-modules), anxiety (STAI), health expectations, demographics and comorbidity. RESULTS: Between diagnostic groups no significant differences were found on generic QL. As compared to the general population, generic QL was impaired on all aspects except bodily pain. Using stepwise regression analysis, 46% of the variance in overall QL was explained with the SF-36 scales vitality (Beta = 0.43) and mental health (Beta = 0.23) being the most important predictors. CONCLUSIONS: The QL of these patients is seriously impaired. In this stage, not cancer-site specific aspects but fatigue and emotions colour their lives. It is recommended to keep the waiting period brief. In addition, suggestions are offered by which physicians might help their patients in alleviating the distress. PMID- 16705726 TI - Expression of survivin and correlation with PCNA in osteosarcoma. AB - BACKGROUND AND OBJECTIVES: Survivin is a new anti-apoptosis gene with reported expression during fetal development and in cancer tissues. Proliferating cell nuclear antigen (PCNA) is essential for the replication of deoxyribonucleic acid (DNA). In this experiment, we examined the expression of survivin and PCNA in human osteosarcoma tissues and investigating whether survivin and PCNA expressions were correlated with histological grades, histological types, and patients' clinical characteristics. METHODS: Sixty-eight osteosarcoma samples were divided into two groups based on the degree of tumor cell differentiation. Immunohistochemical staining was performed to analyze the expression of survinin and PCNA. Four histological types and patients' clinical characteristics were also recorded. RESULTS: There were abundant levels of survivin and PCNA immunoreactivity in the nucleus and/or cytoplasm of the osteosarcoma cells. All cells essentially revealed the cytoplasmic localization and the nuclear signals of survivin in same cases, while PCNA from the majority of cases predominantly showed nuclear expression. Scoring on the percentage of positive cells indicated that survivin expression was significantly associated with the PCNA-labeling index (LI), which was correlated with the histological grades of osteosarcoma (chi2 = 17.86 and chi2 = 23.75, respectively). CONCLUSIONS: The expression of survivin can be used as a useful indicator in grading the malignancy of osteosarcoma. PMID- 16705727 TI - Positive sentinel lymph node: New controversies regarding axillary node dissection. PMID- 16705728 TI - Outcomes of resection of stage IV rectal cancer with mesorectal excision. AB - BACKGROUND: There is no consensus as to the management of the primary rectal cancer in the presence of distant metastasis and data on the outcomes of radical resection in stage IV rectal cancer are limited. This study aims to evaluate the results of resection of rectal cancer in the patients with stage IV disease and to analyze the factors that might affect the survival of these patients. METHODS: Of the 744 patients with radical resection of primary rectal and rectosigmoid cancer during the study period from August 1993 to July 2002, 70 had stage IV disease on the initial presentation. The demographics, the operative details, the tumor characteristics, the postoperative outcomes and survival of the patients were collected prospectively. Factors influencing the survival were analyzed with univariate and multivariate analysis. RESULTS: Fifty-three men and 17 women with a median age of 66 years (range: 31-90 years) were included. The median level of the tumor from the anal verge was 10 cm (range 3-20 cm). The operations included abdominoperineal resection (n = 5), anterior resection (n = 53), and Hartmanns operation (n = 11). The operation mortality was 4.3%. The overall morbidity was 42.7% while the surgical morbidity and the reoperation rates were 15.7% and 5%, respectively. The local recurrence rate was 4.3% and the 2-year actuarial rate was 7.8%. All the patients who had local recurrences also had disseminated peritoneal metastasis. The median cancer-specific survival of the patients who survived the surgery was 15.2 months. Multivariate analysis showed that the presence of gross residual local disease, lymph node metastasis, liver involvement of over 50%, the absence of surgical management of liver metastasis and those without chemotherapy were independent factors associated with poor survival. CONCLUSIONS: Postoperative mortality and morbidity were acceptable in patients with stage IV rectal cancer. The local disease can be controlled effectively with radical resection. However, in patients with extensive liver involvement and advanced local disease, resection is not worthwhile because of the poor survival. Surgical management of the metastasis and the administration of chemotherapy are associated with better survival. However, the optional treatment regimes are yet to be defined. PMID- 16705729 TI - A practical scoring system based upon ROC analysis for evaluating potential lymph nodes metastasis during gastric surgery. AB - BACKGROUND AND OBJECTIVES: Current preoperative N staging does not offer an accurate estimation of lymph node involvement. We establish a new scoring system for predicting N stages to guide a rational lymphadenectomy for gastric cancer. METHODS: Variables correlated with N stages were selected by multivariate stepwise logistic regression analysis. Variables granted the different scores according to the odds ratio (OR). Receiver operating characteristic (ROC) analysis was used to generate scoring ranges from N0 to N3. The agreement between predicted N staging and actual pN classification was analyzed using kappa statistics. RESULTS: Tumor size, depth of invasion, and histological types were selected to establish the scoring system. Scores 0-4, 5-7, 8-9, and 10-13 were postulated to predict N0-3, respectively. The predicted N stage has good agreement with the actual pN classifications. The negative predictive values for N0-3 were 87.0, 86.4, 90.4, and 90.2%; the positive predictive values were 74.7, 62.8, 57.3, and 69.6%, respectively. The accuracy is 82% for N0-1, and 83.7% for N2-3. CONCLUSIONS: The new scoring system can predict the N stage of gastric cancer. With its good negative predictive value, it is possible to minimize the potential hazards of applying a more extensive lymph node dissection than necessary. PMID- 16705730 TI - Effectiveness of radical en-bloc-esophagectomy compared to transhiatal esophagectomy in squamous cell cancer of the esophagus is influenced by nodal micrometastases. AB - PURPOSE: The appropriate surgery for curable squamous cell cancer (SCC) of the esophagus is discussed controversially. PATIENTS AND METHODS: Transthoracic esophagectomy (TTE) with radical en-bloc-lymphadenectomy was performed in 79/120 patients (66%). Due to severe comorbidity, 41/120 patients (34%) underwent less traumatizing transhiatal esophagectomy. Lymph nodes were stained using mAb BER Ep4. RESULTS: pN0 status was found in 54 patients (45%) and pN1 status in 66 patients (55%). In the pN0 subgroup, 30 out of 54 patients (55.6%) had occult tumor cells in lymph nodes detected with BER-Ep4. pN1 patients had additional BER Ep4 positive cells in histopathologically negative lymph nodes in 41 out of 66 patients (59.2%). Median disease-specific survival was 31.4 and 16.8 months for the transthoracic and transhiatal groups, respectively (P = 0.129). pN0 patients lived longer after transthoracic resection (P = 0.0349), and in the subgroup without lymph node micrometastases this effect was even stronger (P = 0.025). In contrast, in pN0 patients with nodal microinvolvement, TTE did not entail a survival benefit when compared with the transhiatal group (P = 0.333). CONCLUSION: TTE results in improved outcome in the absence of lymph node micrometastases. Poor survival in patients with lymphatic micro-dissemination indicates systemic disease and the need for adjuvant treatment. PMID- 16705731 TI - Predictors of invasive disease in breast cancer when core biopsy demonstrates DCIS only. AB - BACKGROUND AND OBJECTIVES: Ductal carcinoma in-situ on core biopsy does not preclude invasive disease within the excision specimen, resulting in the need for further axillary surgery. We sought to identify predictors of invasion when DCIS is present on core biopsy. METHODS: From a database of 895 breast cancer patients, patients with DCIS on core biopsy who had subsequent surgical excision were identified. Their excision pathology and preoperative mammography were correlated with risk of invasion. RESULTS: Patients (93) with a preoperative diagnosis of DCIS on core biopsy were identified. On excision, 31 patients (33%) had invasive carcinoma and 13 patients (14%) had microinvasion. A preoperative finding of calcification only on mammogram was associated with DCIS on excision (P = 0.014), whereas the presence of other mammographic features increased the risk of invasion. Size > or =5 cm on excision pathology was associated with increased risk of invasion (P = 0.002). Forty-eight percent (n = 11/23) of those diagnosed by ultrasound-guided core biopsy had frank invasion on excision compared to 21% (n = 12/57) diagnosed by stereotactic techniques (P = 0.017). CONCLUSION: Mammographic features and tumor size can help predict invasion in patients who have DCIS on core biopsy. Patients who have features other than calcification on mammography or have tumor size > or =5 cm should be considered for a sentinel node biopsy. PMID- 16705732 TI - Adjuvant aromatase inhibitor therapy for early breast cancer: A review of the most recent data. AB - Tamoxifen is the established adjuvant treatment for postmenopausal women with hormone-sensitive early breast cancer. However, the side-effects associated with tamoxifen therapy have prompted a search for safer and potentially more effective endocrine agents. Results from randomized trials of the third-generation aromatase inhibitors, anastrozole, letrozole and exemestane, demonstrating improved efficacy compared with tamoxifen and favorable tolerability profiles, are discussed in this review. PMID- 16705733 TI - Transthoracic vs. transhiatal esophagectomy: Stage migration muddies the water. PMID- 16705734 TI - Primary reconstruction of small mandibular defects by using mandibular remnant. PMID- 16705735 TI - Tobacco extract but not nicotine impairs the mechanical strength of fracture healing in rats. AB - The influence of nicotine and tobacco extract (without nicotine) alone and in combination on and mechanical strength of closed femoral fractures in rats was investigated. One hundred four male Sprague-Dawley rats were divided into four groups receiving: nicotine, tobacco extract, tobacco extract plus nicotine, and saline. One week prior to fracture, osmotic pumps were implanted subcutaneously in all animals to administer nicotine equivalent to the serum level of nicotine observed in a smoker consuming one to two packs of cigarettes daily. An equivalent volume of saline was administered to the control animals. Tobacco extract was administered orally. A closed transverse femoral diaphysial fracture was performed, and stabilized with an intramedullary pin. The fractures were mechanically tested after 21 days of healing. Tobacco extract alone decreased the mechanical strength. Ultimate torque and torque at yield point of the tobacco extract group were decreased by 21% (p=0.010) and 23% (p=0.056), respectively, compared with the vehicle (saline) group, and by 20% (p=0.023) and 26% (p=0.004), respectively, compared with the nicotine group. No difference was found between the tobacco extract and tobacco extract plus nicotine groups. An 18% (p=0.013) reduction in torque at yield point was observed in the tobacco extract plus nicotine group compared with the nicotine group. No differences in ultimate stiffness, energy absorption, and callus bone mineral content at the fracture line were found between any of the groups. Serum levels of nicotine were between 40-50 ng/mL in the group given nicotine alone and the group given tobacco extract plus nicotine (equivalent to serum levels observed in persons smoking one to two packs of cigarettes per day). PMID- 16705736 TI - Stretch-induced modulation of matrix metalloproteinases in mineralizing osteoblasts via extracellular signal-regulated kinase-1/2. AB - Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) produced by osteoblasts play an essential role in bone remodeling. Hence, these proteins could provide an interesting means by which mechanical loading leads to adaptation of bone. Here, we examined the effect of stretch on MMP-1, 2, -3, -8, -9, -13, and -14, as well as TIMP-1 and -2 gene expression in differentiating, mineralizing, and nonmineralizing human SV-40 immortalized preosteoblast cells. In the mineralizing osteoblast culture, but not in the nonmineralizing cultures, cyclic stretch for only 15 min resulted in an increase of MMP-1 (fourfold) and -3 (depending on differentiation stage up to 25-fold) transcript abundance. No clear effect was observed for other MMPs, TIMP-1 or -2. The increase of MMP-1 and -3 was confirmed on the protein level. Stretching experiments performed in the presence of a specific inhibitor of extracellular signal-regulated kinase (ERK) showed a strong suppression of the stretch-induced increase in MMP-1 and -3. In conclusion, we show that MMP-1 and MMP-3 are mechanosensitive genes in mineralizing the human osteoblast, and that the mechano induction of these genes is mediated via the ERK pathway. Our findings implicate that these MMPs are important factors in the mechanoregulation of bone turnover. With the ability to generate MMPs at highly stretched sites, osteoblasts can potantially direct osteoclasts to specific bone surface areas prepared for resorption. PMID- 16705737 TI - Role of polyamines in arginine-dependent colon carcinogenesis in Apc(Min) (/+) mice. AB - We evaluated the role of polyamines in arginine-dependent intestinal tumorigenesis in Apc(Min) (/+) mice. Arginine is a substrate for ornithine synthesis and thus can influence polyamine production. Supplementing the diet with arginine increased intestinal and colonic polyamine levels and colonic carcinogenesis. Inhibiting polyamine synthesis with D,L-alpha diflouromethylornithine (DFMO) decreased small intestinal and colonic polyamine pools. In mice provided basal diet, but not when supplemented with arginine, DFMO decreased small intestinal tumor number and burden, and increased intestinal apoptosis. In mice provided supplemental arginine in the diet, DFMO induced late apoptosis and decreased tumorigenesis in the colon. DFMO slightly reduced tumor incidence, number, and size while significantly decreasing tumor burden and grade. These changes in colon tumorigenesis did not occur in mice not provided supplemental arginine. Our study indicates that polyamines play unique roles in intestinal and colonic carcinogenesis in Apc(Min) (/+) mice. Inhibition of polyamine synthesis suppresses the arginine-dependent risk of colon tumorigenesis, resulting in apoptosis induction and decreased tumorigenesis, in this murine model. PMID- 16705738 TI - Three-dimensional architecture of the left ventricular myocardium. AB - Concepts for ventricular function tend to assume that the majority of the myocardial cells are aligned with their long axes parallel to the epicardial ventricular surface. We aimed to validate the existence of aggregates of myocardial cells orientated with their long axis intruding obliquely between the ventricular epicardial and endocardial surfaces and to quantitate their amount and angulation. To compensate for the changing angle of the long axis of the myocytes relative to the equatorial plane of the ventricles with varying depths within the ventricular walls, the so-called helical angle, we used pairs of cylindrical knives of different diameters to punch semicircular slices from the left ventricular wall of pigs, the slices extending from the epicardium to the endocardium. The slices were pinned flat, fixed in formaldehyde, embedded in paraffin, sectioned, stained with azan or hematoxilin and eosin, and analyzed by a new semiautomatic procedure. We made use of new techniques in informatics to determine the number and angulation of the aggregates of myocardial cells cut in their long axis. The alignment of the myocytes cut longitudinally varied markedly between the epicardium and the endocardium. Populations of myocytes, arranged in strands, diverge by varying angles from the epicardial surface. When paired knives of decreasing diameter were used to cut the slices, the inclination of the diagonal created by the arrays increases, while the lengths of the array of cells cut axially decreases. The visualization of the size, shape, and alignment of the myocytic arrays at any side of the ventricular wall is determined by the radius of the knives used, the range of helical angles subtended by the alignment of the myocytes throughout the thickness of the wall, and their angulation relative to the epicardial surface. Far from the majority of the ventricular myocytes being aligned at angles more or less tangential to the epicardial lining, we found that three-fifths of the myocardial cells had their long axes diverging at angles between 7.5 and 37.5 degrees from an alignment parallel to the epicardium. This arrangement, with the individual myocytes supported by connective tissue, might control the cyclic rearrangement of the myocardial fibers. This could serve as an important control of both ventricular mural thickening and intracavitary shape. PMID- 16705739 TI - Monocyte chemotactic protein-1 (MCP-1) acts as a paracrine and autocrine factor for prostate cancer growth and invasion. AB - BACKGROUND: Monocyte chemotactic protein-1 (MCP-1) plays a key role in the recruitment and activation of monocytes during inflammation. Increased MCP-1 serum levels in patients with various cancers were correlated with advanced stage. Here, we evaluated the role of MCP-1 on prostate cancer (CaP) cell proliferation and invasion. METHODS: Expression of MCP-1 in tissue specimens was analyzed by immunohistochemical staining. MCP-1 production was determined by ELISA in conditioned media collected from primary prostate epithelia (PrEC), LNCaP, C4-2B, PC3 cells, and hFOB. Cell proliferation and invasion were assayed by MTS assay and invasion chambers. RESULTS: All CaP cells, as well as hFOB, produced high amount of MCP-1 compared to PrEC cells. MCP-1 expression levels were associated with advanced pathologic stage. MCP-1 induced proliferation and invasion of CaP cells and this was abolished partially either by CCR2 antagonist or PI3 Kinase inhibitor. CONCLUSION: MCP-1 acts as a paracrine and autocrine factor for CaP growth and invasion. PMID- 16705740 TI - An androgen-independent androgen receptor function protects from inositol hexakisphosphate toxicity in the PC3/PC3(AR) prostate cancer cell lines. AB - BACKGROUND: Inositol hexakisphosphate (IP6) is a phytochemical exhibiting anticancer activity. Because few prostate cancer (PCa) cell lines have been used to study IP6, we assessed its efficacy in a panel of PCa cell lines. METHODS AND RESULTS: Using WST-1 assays we observed that, although androgens did not modulate its efficacy, IP6 was more active in androgen receptor (AR) negative cells than in AR-positive cells. Stable expression of the AR in PC3 cells (PC3(AR)) decreased the response to IP6, which was reversed by an AR-targeting siRNA. Furthermore, AR expression in PC3 cells resulted in significantly reduced caspase 3 activation (P < 0.001) and DNA fragmentation (P < 0.05) in response to IP6. Similarly, although treatment with IP6 caused the upregulation of NF-kappaB responsive (IkappaB-alpha, IRF-2) and p53/E2F-responsive genes (Puma, Noxa) in PC3 cells, this increase was reduced in PC3AR cells (P < 0.01). CONCLUSION: We conclude that resistance to IP6 can be linked to a ligand-independent AR function. PMID- 16705741 TI - Detection of human herpesvirus 8 (HHV-8) in normal prostates. AB - BACKGROUND: Human herpesvirus 8 (HHV-8) DNA has been detected in semen and prostatic tissues in some, but not all reports. We have analyzed prostate tissues from HHV-8 seropositive men for the expression of viral proteins and determined if expression of these proteins are associated with increased inflammation. METHODS: Paraffin sections of non-cancerous prostates from HHV-8 seropositive (n = 16) and seronegative (n = 2) men who died with AIDS were screened for expression of three viral proteins by immunohistochemistry. Levels of inflammation were determined by expression of CD68 and CD20. Cellular proliferation was determined by expression of Ki67. RESULTS: Among the 16 HHV-8 seropositive cases, 68.9% (11/16) (95% C.I. = 0.41-0.89) were positive for HHV-8 protein expression, while the 2 seronegative patients showed no HHV-8 protein expression. There was increased inflammation among HHV-8 positive prostates. CONCLUSIONS: These results demonstrate that HHV-8 is present in normal prostates of HIV-infected men and the expression of viral proteins is associated with increased localized inflammation. PMID- 16705742 TI - Proteomic comparison of prostate cancer cell lines LNCaP-FGC and LNCaP-r reveals heatshock protein 60 as a marker for prostate malignancy. AB - BACKGROUND: Androgen-sensitive prostate cancer cell-line LNCaP-FGC and androgen resistant line LNCaP-r constitute a model for development of androgen resistance in prostate cancer. METHODS: Proteins differently expressed in the two cell-lines were identified by two-dimensional (2-D) electrophoresis and mass spectrometry. HSP60, more abundant in LNCaP-r, was studied by RT-PCR and immunohistochemistry in specimens of human prostate cancer. RESULTS: HSP60 was upregulated in LNCaP-r, nm23 in LNCaP-FGC, and titin (two isoforms) in either LNCaP-r or LNCaP-FGC. In non-malignant prostate, HSP60-staining was in the glandular compartment, particularly basal epithelial cells. In prostate cancer, most epithelial cells showed moderate-strong staining without apparent correlation between staining intensity and Gleason grade. CONCLUSIONS: The LNCaP-FGC/LNCaP-r model, characterized by 2-D electrophoresis, reveals distinct proteomic alterations. With HSP60, results from cell-lines correlated with clinical results, indicating that this model can be used for dissection of mechanisms involved in transformation to androgen resistance and assignment of protein markers in prostate cancer. PMID- 16705743 TI - Involvement of arginine methyltransferase CARM1 in androgen receptor function and prostate cancer cell viability. AB - BACKGROUND: Androgen receptor (AR) may play a role in prostate cancer progression. Coactivator-associated arginine methyltransferase (CARM1) catalyzes methylation of histone H3 at Arg-17. METHODS: Immunohistochemistry of CARM1 was performed on primary prostate cancer specimens. CARM1 recruitment and histone methylation was analyzed by chromatin immunoprecipitation. The effect of CARM1 overexpression or CARM1 knockdown was assessed on reporter assays, cell proliferation, apoptosis, and endogenous androgen target gene expression. RESULTS: CARM1 expression was increased in the nucleus of castration-resistant, but not androgen-stimulated prostate cancer. Androgen stimulation led to CARM1 recruitment and methylation of histone H3 at androgen responsive enhancers. Overexpression of CARM1 stimulated and CARM1 knockdown inhibited AR reporter activity. CARM1 knockdown inhibited cell proliferation and induced apoptosis. CARM1 knockdown inhibited androgen-dependent prostate specific antigen (PSA) and hK2 mRNA expression. CONCLUSIONS: CARM1 is essential for AR function and may play a role in prostate cancer progression. CARM1 may represent a novel therapeutic target in prostate cancer. PMID- 16705744 TI - Changes in gene expressions elicited by physiological concentrations of genistein on human endometrial cancer cells. AB - The incidence of hormone-related diseases such as prostatic, breast, ovarian, and endometrial cancer is lower in Asian populations compared to Western countries. High consumption of soybean products that are rich in phytoestrogens, predominantly genistein, is postulated to be responsible for the lower incidence of hormone-related disease, although the mechanism through which this effect might be mediated is unclear. In this study, microarray analysis was used to identify the changes in gene expression elicited by treatment of the human endometrial cancer cell line, Ishikawa, with genistein at both physiologically achievable and supraphysiological concentrations. Genistein treatment at 5 microM concentration induced multiple changes in gene expression including some implicated in oncogenesis. In contrast, treatment with a supraphysiological concentration of genistein predominantly activated stress response genes and showed very limited overlap with the genes regulated at lower concentrations. Of the genes regulated by genistein, 9.3% were also regulated by 17beta-estradiol suggesting that genistein exerts its response via the estrogen pathway. These results indicate that at physiological concentrations, genistein is able to elicit pleiotropic effects on a variety of pathways believed to be involved in tumorigenesis. Supraphysiological concentrations of genistein, such as those used in many previous studies, elicit changes in gene expression that are unlikely to occur in vivo. PMID- 16705745 TI - A bidirectional "alpha(v)beta(3) integrin-ERK1/ERK2 MAPK" connection regulates the proliferation of breast cancer cells. AB - In addition to their role in cell migration and adhesion, integrins elicit a series of transduction events that regulate cell-cycle progression and apoptosis in a process known as "outside-in" signaling. A second mode of integrin regulation known as "inside-out" signaling, in which the activation of major cell transduction cascades can influence the activation status of some integrins, has also been described. Here, we have assessed the role of the extracellular signal regulated kinase (ERK1)/ERK2, mitogen-activated protein kinase (MAPK), and phospoinositide 3-kinase (PI-3'K) signaling pathways in the expression and function of alpha(v)beta(3) integrin in breast cancer models. Pharmacological inhibition of MEK1 and MEK2 with U0126 drastically increased the levels of alpha(v)beta(3) in Heregulin (HRG)-overexpressing MDA-MB-231 cells (231/WT, 231/VEC) and derivatives transfected with the antisense orientation of the HRG beta2 full length cDNA (231/ASPOOL, 231/AS31). Interestingly, this was related to a significant decrease of viability and of the S- and G2/M subcompartment of the cell cycle in MDA MB 231 cells in response to U0126. Furthermore, specific inhibition of the PI-3'K pathway with LY294002 also induced an increase of alpha(v)beta(3) levels but to a lesser extent. Moreover, pretreatment of MDA-MB 231 cells with U0126 antagonized the effects of small peptidomimetic alpha(v)beta(3) antagonists. Remarkably, inhibition of the PI-3'K/AKT pathway did not exert the same effects, thus suggesting that the "outside-in" as well as the "inside-out" alpha(v)beta(3)-mediated signaling goes primarily through the ERK1/ERK2 MAPK pathway in MDA MB 231 breast cancer cells. Collectively, these results strongly suggest the existence of a bidirectional molecular connection alpha(v)beta(3)-ERK1/ERK2 MAPK that would regulate breast cancer cells survival and proliferation. PMID- 16705747 TI - DNA-templated synthesis in three dimensions: Introducing a micellar scaffold for organic reactions. PMID- 16705748 TI - Identification of phytochrome-interacting protein candidates in Arabidopsis thaliana by co-immunoprecipitation coupled with MALDI-TOF MS. AB - Phytochrome-interacting proteins have been extensively studied to elucidate light signaling pathway in plants. However, most of these proteins have been identified by yeast two-hybrid screening using the C-terminal domain of phytochromes. We used co-immunoprecipitation followed by proteomic analysis in plant cell extracts in an attempt to screen for proteins interacting either directly or indirectly with native holophytochromes including the N-terminal domain as well as C terminal domain. A total of 16 protein candidates were identified, and were selected from 2-DE experiments. Using MALDI-TOF MS analysis, 7 of these candidates were predicted to be putative phytochrome A-interacting proteins and the remaining ones to be phytochrome B-interacting proteins. Among these putative interacting proteins, protein phosphatase type 2C (PP2C) and a 66-kDa protein were strong candidates as novel phytochrome-interacting proteins, as knockout mutants for the genes encoding these two proteins had impaired light-signaling functions. A transgenic knockout Arabidopsis study showed that a 66-kDa protein candidate regulates hypocotyl elongation in a light-specific manner, and altered cotyledon development under white light during early developmental stages. The PP2C knockout plants also displayed light-specific changes in hypocotyl elongation. These results suggest that co-immunoprecipitation, followed by proteomic analysis, is a useful method for identifying novel interacting proteins and determining real protein-protein interactions in the cell. PMID- 16705749 TI - Proteomic analysis of Tityus discrepans scorpion venom and amino acid sequence of novel toxins. AB - The Venezuelan scorpion Tityus discrepans is known to cause human fatalities. We describe the first complete proteomic analysis of its venom. By HPLC 58 different fractions were obtained and 205 different components were identified by MS analysis. Components having molecular masses from 272 to 57 908 amu were found. Forty homogeneous components had their N-terminal amino acid sequence determined by Edman degradation, from which two new peptides named TdK2 and TdK3 (meaning T. discrepans (Td) K(+) channel toxins 2 and 3) were fully characterized. The first contains 34 amino acid residues with a molecular mass of 3451 amu, and the second has 36 amino acids with 3832 amu. Both peptides are tightly bound by three disulfide bridges. TdK2 was shown to block reversibly the Shaker B K(+)-channel expressed heterologously in Sf9 cells. The systematic number assigned to TdK2 is alpha-KTx-18.2 and that of TdK3 is alpha-KTx-18.3. Comparative analysis of the amino acid sequences found suggests that this venom contains peptides highly similar to those that block K(+) channels, as well as those that modify the gating mechanisms of Na(+) channels, found in other scorpions. Additionally, peptides similar to defensins were also identified. PMID- 16705750 TI - Subcellular pH and predicted pH-dependent features of proteins. AB - A characteristic of two-dimensional proteomics gels is a general bimodal distribution of isoelectric (pI) values. Discussion of this feature has focussed on the balance of acidic and basic ionisable residues, and potential relationships between pI distributions and organism classification or protein subcellular location. Electrostatics calculations on a set of protein structures with known subcellular location show that predicted folded state pI are similar to those calculated from sequence alone, but adjusted according to a general stabilising effect from interactions between ionisable groups. Bimodal distributions dominate both pI and the predicted pH of maximal stability. However, there are significant differences between these features. The average pH of maximal stability generally follows organelle pH. Average pI values are well removed from organelle pH in most subcellular environments, consistent with the view that proteins have evolved to carry (on average) net charge in a given subcellular location, and relevant to discussion of solubility in crowded environments. Correlation of the predicted pH of maximum stability with subcellular pH suggests an evolutionary pressure to adjust folded state interactions according to environment. Finally, our analysis of ionisable group contributions to stability suggests that Golgi proteins have the largest such term, although this dataset is small. PMID- 16705751 TI - Proteomic dissection of potential signal recognition particle dependence in protein secretion by Bacillus subtilis. AB - The bacterial signal recognition particle (SRP)-dependent pathway is believed to be a major targeting route for membrane proteins, as well as for subsets of secretory proteins. The present studies were aimed at an assessment of the role of two key components of SRP, namely Ffh and FtsY, in protein secretion by the Gram-positive bacterium Bacillus subtilis. Our results show that both components are important for the extracellular accumulation of proteins containing known signal peptides. Remarkably, extracellular accumulation of individual proteins was affected to different extents by depletion of Ffh or FtsY, at least under the conditions tested. Moreover, the observed Ffh or FtsY dependence of certain secretory proteins did not seem to correlate with signal peptide length or hydrophobicity. Although it is presently difficult to distinguish between direct and indirect effects, these findings suggest that other, yet unidentified, determinants in secretory proteins are also important for their SRP dependence. High-level production of homologous and heterologous secretory proteins was shown to result in elevated cellular Ffh and FtsY levels. This phenomenon is, most likely, due to post-transcriptional regulation. In conclusion, the present proteomic dissection of SRP-dependent extracellular protein accumulation provides exciting leads to identify novel determinants for interactions between secretory proteins and SRP. PMID- 16705752 TI - The phosphate-starvation response of Bacillus licheniformis. AB - The phosphate-starvation stimulon of Bacillus licheniformis was analyzed at the transcriptional and translational level. The comparison of the transcriptome and the proteome demonstrated that this specific starvation response of B. licheniformis is partially similar to that of B. subtilis. However, it is also shown that B. licheniformis has evolved its own strategies to cope with this nutrient limitation. By means of the secretome analysis the phytase was identified as the most abundant protein under phosphate-starvation conditions. Data of this study indicate that, unlike in B. subtilis, phosphate starvation in B. licheniformis does not induce the SigmaB-dependent general stress response. PMID- 16705753 TI - A combined proteomic and transcriptomic approach to the study of stage differentiation in Leishmania infantum. AB - Protozoan parasites of the genus Leishmania are found as promastigotes in the sandfly vector and as amastigotes in mammalian macrophages. Mechanisms controlling stage-regulated gene expression in these organisms are poorly understood. Here, we applied a comprehensive approach consisting of protein prefractionation, global proteomics and targeted DNA microarray analysis to the study of stage differentiation in Leishmania. By excluding some abundant structural proteins and reducing complexity, we detected and identified numerous novel differentially expressed protein isoforms in L. infantum. Using 2-D gels, over 2200 protein isoforms were visualized in each developmental stage. Of these, 6.1% were strongly increased or appeared unique in the promastigote stage, while the relative amounts of 12.4% were increased in amastigotes. Amastigote-specific protein isoform and mRNA expression trends correlated modestly (53%), while no correlation was found for promastigote-specific spots. Even where direction of regulation was similar, fold-changes were more modest at the RNA than protein level. Many proteins were present in multiple spots, suggesting that PTM is extensive in this organism. In several cases, different isoforms appeared to be specific to different life stages. Our results suggest that post-transcriptional controls at translational and post-translational levels could play major roles in differentiation in Leishmania parasites. PMID- 16705754 TI - A proteomic study on human osteoblastic cells proliferation and differentiation. AB - Changes in expression profiles for 17 proteins were ascertained in human mature osteoblasts compared to pre-osteoblasts (differentiation markers). A differential approach was used to highlight proteomic changes between human osteosarcoma cells and mature osteoblasts, showing a relative over-expression of 8 proteins (proliferation and tumor indicators), as well as under-expression of proteins also found down-regulated in pre-osteoblasts (specific markers of osteoblast differentiation). Our findings confirmed the differences between cell lines and primary human cell cultures and suggested caution on the use of osteosarcoma to study anti-osteoporotic drugs in humans. PMID- 16705755 TI - Serum proteome profiles identifies parathyroid hormone physiologic response. AB - Parathyroid hormone (amino acids 1-34) (PTH) regulates bone and calcium homeostasis. The magnitude of the effects of PTH on bone varies in osteoporosis patients. We employed ProteinChip technology to generate protein profiles from sera of mice treated once daily with PTH or vehicle for 3 or 11 days. Data analyses on selected arrays indicated significant increases in serum proteins or peptides in PTH-treated groups, compared to vehicle-controls. The magnitude of change increased with duration of treatment. Anion-exchange fractionation of sera prior to profiling on array surfaces increased the number of proteins detected that were regulated by PTH. The optimized purification conditions developed "on chip" for subsets of proteins, reflected corresponding behavior with process compatible chromatographic resins under elution chromatography. We have identified and evaluated subsets of serum proteins regulated by PTH treatment, using a combination of ProteinChip technology, column chromatography, PAGE and LC MS/MS. Our data demonstrate the feasibility of using a panel of serum proteins to detect PTH responsiveness in humans. PMID- 16705756 TI - Inducible nitric oxide synthase is not essential for the development of fibrosis and liver damage induced by CCl4 in mice. AB - The aim of the present work was to investigate the role of inducible nitric oxide (NO) synthase (iNOS) in CCl(4)-induced cirrhosis by utilizing iNOS knock out mice (iNOS(-/-)). Cirrhosis was produced by i.p. administration of CCl(4) (1 ml kg(-1) of body weight) dissolved in olive oil three times a week for 3 months to iNOS(-/ ) or iNOS(+/+) (wild type) mice; appropriate olive oil controls were performed. Nitrite plus nitrate levels were lower in iNOS(-/-) compared with iNOS(+/+) mice, but CCl(4) did not produce a significant effect in any mice. Reduced (GSH) glutathione was increased in iNOS(-/-) mice receiving vehicle and in both groups receiving CCl(4); lipid peroxidation increased significantly in iNOS(+/+) but not in iNOS(-/-) mice. Bilirubins, alanine aminotransferase and collagen (measured as the hepatic hydroxyproline content) were increased significantly by the chronic intoxication with CCl(4) in both iNOS(-/-) and iNOS(+/+) mice; importantly there was no difference between these groups. This study clearly suggests that NO derived from iNOS does not participate in cholestasis, necrosis or fibrosis induced by CCl(4) in the mice. The present results are in disagreement with several studies indicating a beneficial or detrimental effect of this molecule utilizing different experimental approaches and in agreement with some studies indicating that NO does not affect liver damage in some models. It must be pointed out that this is the first report in iNOS knock out mice utilizing the chronic model of intoxication with CCl(4); thus, comparisons with other models or approaches are difficult to reconcile. PMID- 16705757 TI - Evaluation of an ex vivo murine local lymph node assay: multiple endpoint comparison. AB - The local lymph node assay (LLNA) is used to assess the skin sensitization potential of chemicals. In the standard assay, mice are treated topically on the dorsum of both ears with test substance for 3 days. Following 2 days of rest, the initiation of the hypersensitivity response is evaluated by injecting (3)H thymidine into a tail vein, and then measuring the levels of radioisotope incorporated into the DNA of lymph node cells draining the ears. In the current study, BALB/c mice were treated with the contact sensitizers hexylcinnamic aldehyde (HCA) and oxazolone, and the nonsensitizer methyl salicylate. The proliferative response of lymph node cells was evaluated in an ex vivo assay, in which isolated cells were cultured in vitro with (3)H-thymidine. Treatment of mice with HCA at 5-50% resulted in concentration-related increases in (3)H thymidine incorporation, with stimulation indices ranging from 3 to 14. Low animal-to-animal variability was seen in three replicate assays testing HCA at 25%. As anticipated, the proliferative response induced by the potent sensitizer oxazolone at 0.25% was greater than HCA at all concentrations tested. Stimulation indices of 1.5 and 3 were seen in two independent experiments with methyl salicylate. These equivocal findings were likely due to the irritancy properties of the compound. Importantly, measuring ex vivo (3)H-thymidine incorporation was more sensitive than evaluating lymph node weight and cellularity, and in vitro bromodeoxyuridine incorporation. Furthermore, the results of the ex vivo LLNA were comparable to the standard assay. This study provided evidence that supports the use of an ex vivo LLNA for hazard assessment of contact hypersensitivity. PMID- 16705759 TI - Proceedings of the International Embryo Transfer Society (IETS) Pre- and Post Conference Symposia, Orlando, Florida, USA, January 2006. PMID- 16705758 TI - Spotlight on Silvia Priori, MD. PMID- 16705760 TI - Food & Drug Administration panel approves heart failure medication aimed at African-Americans. PMID- 16705761 TI - The making of environmental law. PMID- 16705762 TI - The case of the female orgasm. PMID- 16705763 TI - Response to clinical skills at conferences. PMID- 16705764 TI - Comment on ILCA's official response to the AAP'S SIDS statement. PMID- 16705765 TI - Response to ILCA's official response to the AAP's SIDS statement. PMID- 16705766 TI - Depression among students on the rise. PMID- 16705767 TI - AVMA answers: parting thoughts from Dr. Carlson. Interview by Katie Burns. PMID- 16705768 TI - Veterinary academe evaluates animal use in education. PMID- 16705769 TI - Offers feedback on JAVMA features. PMID- 16705770 TI - Interest in turtle-associated salmonellosis piqued. PMID- 16705771 TI - Course of anomalous left main not clearly determined by angiography. PMID- 16705772 TI - Dental and legal considerations in periodontal therapy. PMID- 16705773 TI - Fecal occult blood testing: Forget the finger! PMID- 16705774 TI - Anti-platelet therapy and recurrent ulcers: Rethinking rebleeding. PMID- 16705775 TI - Sacral spinal nerve stimulation for fecal incontinence: A viable therapeutic option for refractory incontinence. PMID- 16705776 TI - Delayed treatment of acute hepatitis HCV? Yes, but when, who, and how? PMID- 16705777 TI - Off-label reports of new biologics: exciting new therapy or dubious research? Examples from recombinant activated factor VII. PMID- 16705778 TI - Grandparent and relative caregivers: medical consent and educational enrollment laws. PMID- 16705779 TI - Outbreak of trichinellosis in French hunters who ate Canadian black bear meat. PMID- 16705780 TI - More on novel oxygen-concentrator-based equipment; part 1. PMID- 16705782 TI - Proceedings of the 2nd Strategies for Engineered Negligible Senescence (SENS) Conference, Part 1. Cambridge, United Kingdom, September 2005. PMID- 16705781 TI - More on novel oxygen-concentrator-based equipment; part 2. PMID- 16705784 TI - Introduction: those who do not look ahead, stay behind. PMID- 16705783 TI - [A suspected case of sarcoidosis, developing obstructive jaundice 2 years later]. PMID- 16705785 TI - Child maltreatment in American Indian and Alaska Native communities: integrating culture, history, and public health for intervention and prevention. AB - This article addresses child maltreatment intervention and prevention among American Indians and Alaska Natives. The authors argue that history and culture must be included as context and variables for developing and implementing prevention programs in Indian Country. They propose that the public health violence prevention model would benefit from incorporating tenets of the history and culture(s) of diverse groups, in this instance American Indians and Alaska Natives. The authors offer an approach that focuses on population- and individual level risk and protective factors for child maltreatment intervention and prevention in American Indian/Alaska Native communities. They include suggestions and examples for doing the work in Indian Country. PMID- 16705786 TI - Views of child sexual abuse in two cultural communities: an exploratory study among African Americans and Latinos. AB - This exploratory study investigates knowledge and ideas about child sexual abuse among African Americans and Latinos through focus group discussions. Participants defined and described child sexual abuse, acknowledged that it occurred in their communities, and expressed their sense that family risk factors, risky institutions, and offender propensities were its root causes. Latino participants identified cultural transitions as another contributor. Responses and conversational style differed somewhat by gender and cultural identity. The authors discuss implications for child sexual abuse prevention, intervention, and research. PMID- 16705787 TI - Ethnicity, income, and parenting contexts of physical punishment in a national sample of families with young children. AB - Nearly half of U.S. parents use physical punishment for child discipline. Although some studies relate physical punishment and later dysfunction, others suggest that its effects depend on the context in which it is used. The authors analyzed data from the Commonwealth Fund Survey of Parents With Young Children, a national sample of 2,017 parents with children younger than 3. Parents reported their use of spanking, five other disciplinary practices, and four nurturing interactions. The authors used cluster analysis to define four groups of parents with distinct patterns of discipline and nurturing. Two groups with above-average use of spanking shared a high prevalence of parent depressive symptoms and a low level of nurturing but had markedly different demographic profiles and use of nonphysical punishment. Parents who used average levels of spanking made frequent use of nonphysical disciplinary strategies and had high levels of nurturing interactions. Parents who reported below-average spanking had relatively low levels of both disciplinary and nurturing interactions. PMID- 16705788 TI - The relationship between familism and child maltreatment in Latino and Anglo families. AB - Familism, or familismo, refers to attitudes, behaviors, and family structures operating within an extended family system and is believed to be the most important factor influencing the lives of Latinos. Because of the complexity of the construct, this article begins by separating out and defining each dimension of familism, and then clarifies its relationship to the broader literature on social networks, social support, and child maltreatment. The analysis tests whether each dimension of familism is related to child maltreatment within and between 35 abusive Latino, 35 nonabusive Latino, 51 abusive Anglo, and 51 nonabusive Anglo families. Nonabusing Latinas appear to have a higher level of familism than the other three groups of mothers. However; when intraethnic comparisons were made, nonabusive Anglos, compared with abusive Anglos, had higher levels of familism on several variables. Hence, familism seems to characterize families--Latino and Anglo--who do not maltreat their children. PMID- 16705789 TI - Ethnicity in child maltreatment research: a content analysis. AB - The present study represents a content analysis of 1,133 articles published between 1977 and 1998 in Child Abuse and Neglect, Child Maltreatment, and Journal of Child Sexual Abuse was performed. Although there were indications of increased attention to ethnicity in child maltreatment research published between 1977 and 1998, the improvements were modest. Of the 1,133 articles examined, 6.7% focused on ethnicity. The percentage of studies that provided information on the ethnic composition of samples increased across time; however, only half of the studies published in the most recent years examined (1995-1998) reported information on the ethnicity of the participants. Although the percentage of articles that used ethnicity in analyses or as a control variable increased across time, three quarters of the articles published in the most recent years examined (1995-1998) did not include ethnicity in their analyses or design. PMID- 16705790 TI - The importance of culture in treating abused and neglected children: an empirical review. AB - There is growing evidence that cultural factors may influence symptom development and treatment referral patterns among abused and neglected children. To date, few treatment outcome studies have specifically examined the impact of race, culture, or ethnicity on treatment response among maltreated children. Those that have attempted to include these factors have typically suffered from lack of clarity of the meaning of these terms. This article reviews the available empirical evidence that addresses the influence of culture on symptom formation, treatment seeking behaviors, treatment preference, and response following child maltreatment. Hypotheses regarding these findings are addressed, and implications for practice, research, and public policy are discussed. PMID- 16705791 TI - A reporting and response model for culture and child maltreatment. AB - As our society becomes increasingly culturally diverse, there is a growing concern in the mental health field as to whether clinicians are able to accurately distinguish different cultural parenting discipline practices from child maltreatment. Clinicians in various fields continue to differ on what is reportable. Although there is literature describing characteristics of various ethnic minority groups, there are limited data to support why clinicians do what they do and no decision-making model to guide a clinician's reporting behavior when working with clients from different cultures. This article focuses on cultural discipline practices rather than healing practices (e.g., coining) that may be challenging to assess. The authors propose a model to guide clinicians through the decision-making process and discuss interventions and clinical responses that may be most appropriate when presented with different scenarios involving cultural parenting discipline practices and child maltreatment. Finally, limitations of the presented model along with future clinical and research directions are discussed. PMID- 16705792 TI - The emerging problem of physical child abuse in South Korea. AB - South Korea has had remarkably high incidence and prevalence rates of physical violence against children, yet the problem has received only limited public and professional attention until very recently. This article represents the first attempt in English to systematically analyze South Korea's recent epidemiological studies on child maltreatment. Discussed are sociocultural factors that have contributed both to delays in child protection laws and a low public awareness of the problem of child abuse. The article highlights methodological issues concerning the definition of physical abuse in South Korea and the complex attitudes toward violence. It also examines the role of the Korean women's movement in the reform of family laws and the recent establishment of new child protection legislation. Suggestions for future directions for the problem of child maltreatment within South Korea are presented. PMID- 16705794 TI - First International Symposium on Recent Advances in Environmental Health Research. PMID- 16705793 TI - Ethnic differences in risk behaviors and related psychosocial variables among a cohort of maltreated adolescents in foster care. AB - This study examined the cross-ethnic equivalence of measures and the relationships between psychosocial variables and risk behaviors in an ethnically diverse sample of maltreated adolescents 6 years after their placement in foster care. Overall, there was cross-ethnic measurement equivalence, except for the self-destructive behavior and perceived opportunities constructs, which did not demonstrate internal consistency for African American youth. The authors found few differences between White (non-Latino), Hispanic, and African American youth on levels of engagement in risk behaviors and across domains of psychosocial functioning. The relationships between the psychosocial variables and risk behaviors were then examined across ethnic groups. The pattern of results was different as a function of ethnicity, as fewer of the psychosocial variables were significantly related to the risk behavior variables for African American youth. Possible explanations for these differences are presented and implications for intervention discussed. PMID- 16705795 TI - Systems biology: new approaches to old environmental health problems. AB - The environment plays a pivotal role as a human health determinant and presence of hazardous pollutants in the environment is often implicated in human disease. That pollutants cause human diseases however is often controversial because data connecting exposure to environmental hazards and human diseases are not well defined, except for some cancers and syndromes such as asthma. Understanding the complex nature of human-environment interactions and the role they play in determining the state of human health is one of the more compelling problems in public health. We are becoming more aware that the reductionist approach promulgated by current methods has not, and will not yield answers to the broad questions of population health risk analysis. If substantive applications of environment-gene interactions are to be made, it is important to move to a systems level approach, to take advantage of epidemiology and molecular genomic advances. Systems biology is the integration of genomics, transcriptomics, proteomics, and metabolomics together with computer technology approaches to elucidate environmentally caused disease in humans. We discuss the applications of environmental systems biology as a route to solution of environmental health problems. PMID- 16705796 TI - Down-regulation of the expression of the FIH-1 and ARD-1 genes at the transcriptional level by nickel and cobalt in the human lung adenocarcinoma A549 cell line. AB - Although nickel and cobalt compounds have been known to cause induction of the transcription factor hypoxia-inducible factor 1 (HIF-1) and activation of a battery of hypoxia-inducible genes in the cell, the molecular mechanisms of this induction remain unclear. The post-translational modification of HIF-1a, the oxygen-sensitive subunit of HIF-1, regulates stabilization, nuclear translocation, DNA binding activity, and transcriptional activity of the protein. Among the enzymes regulating the post-translational modification of HIF-la, the factor inhibiting HIF-1 (FIH-1) hydroxylates the protein at asparagine 803, suppressing the interaction of HIF-1a with transcription coactivators p300/CBP and reducing the transcriptional activity of the protein. ARD-1, the acetyltransferase, acetylates HIF-1a at lysine 532, which enhances the interaction of HIF-1a with pVHL. Therefore, FIH-1 and ARD-1 negatively regulate the transcriptional activity and the stability of HIF-1a. We examined the mRNA levels of FIH-l and ARD-1 genes after exposure nickel (II) or cobalt (II) to the cell and found that both genes were down-regulated by the chemical treatment, which may lead to reduced levels of both proteins and result in increased level of HIF-1 a and its transcriptional activity. PMID- 16705797 TI - Heavy metals stimulate human LINE-1 retrotransposition. AB - L1 and Alu elements are among the most active retroposons (mobile elements) in the human genome. Several human diseases, including certain forms of breast cancer and leukemia, are associated with L1 and Alu insertions in functionally important areas of the genome. We present data demonstrating that environmental pollutants, such as heavy metals, can stimulate L1 retrotransposition in a tissue culture system using two different types of assays. The response to these agents was equivalent when using a cell line with a stably integrated L1 vector (genomic) or a by introducing the L1 vector by transient transfection (episomal) of the cell. Reproducible results showed that mercury (HgS), cadmium (CdS), and nickel (NiO) increase the activity of L1 by an average of three (3) fold p<0.001. This observation is the first to link several carcinogenic agents with the increased retrotransposition activity of L1 as an alternate mechanism of generating genomic instability contributing to the process of carcinogenesis. Our results demonstrate that mobile element activation must be considered as one of the mechanisms when evaluating genomic damage/instability in response to environmental agents. PMID- 16705798 TI - Induced mitogenic activity in AML-12 mouse hepatocytes exposed to low-dose ultra wideband electromagnetic radiation. AB - Ultra-wideband (UWB) technology has increased with the use of various civilian and military applications. In the present study, we hypothesized that low-dose UWB electromagnetic radiation (UWBR) could elicit a mitogenic effect in AML-12 mouse hepatocytes, in vitro. To test this hypothesis, we exposed AML-12 mouse hepatocytes, to UWBR in a specially constructed gigahertz transverse electromagnetic mode (GTEM) cell. Cells were exposed to UWBR for 2 h at a temperature of 23 degrees C, a pulse width of 10 ns, a repetition rate of 1 kHz, and field strength of 5-20 kV/m. UWB pulses were triggered by an external pulse generator for UWBR exposure but were not triggered for the sham exposure. We performed an MTT Assay to assess cell viability for UWBR-treated and sham-exposed hepatocytes. Data from viability studies indicated a time-related increase in hepatocytes at time intervals from 8-24 h post exposure. UWBR exerted a statistically significant (p < 0.05) dose-dependent response in cell viability in both serum-treated and serum free medium (SFM) -treated hepatocytes. Western blot analysis of hepatocyte lysates demonstrated that cyclin A protein was induced in hepatocytes, suggesting that increased MTT activity after UWBR exposure was due to cell proliferation. This study indicates that UWBR has a mitogenic effect on AML-12 mouse hepatocytes and implicates a possible role for UWBR in hepatocarcinoma. PMID- 16705799 TI - Cytotoxicity assessment of some carbon nanotubes and related carbon nanoparticle aggregates and the implications for anthropogenic carbon nanotube aggregates in the environment. AB - Nanotechnology and nanomaterials have become the new frontier world-wide over the past few years and prospects for the production and novel uses of large quantities of carbon nanotubes in particular are becoming an increasing reality. Correspondingly, the potential health risks for these and other nanoparticulate materials have been of considerable concern. Toxicological studies, while sparse, have been concerned with virtually uncharacterized, single wall carbon nanotubes, and the conclusions have been conflicting and uncertain. In this research we performed viability assays on a murine lung macrophage cell line to assess the comparative cytotoxicity of commercial, single wall carbon nanotubes (ropes) and two different multiwall carbon nanotube samples; utilizing chrysotile asbestos nanotubes and black carbon nanoaggregates as toxicity standards. These nanotube materials were completely characterized by transmission electron microscopy and observed to be aggregates ranging from 1 to 2 microm in mean diameter, with closed ends. The cytotoxicity data indicated a strong concentration relationship and toxicity for all the carbon nanotube materials relative to the asbestos nanotubes and black carbon. A commercial multiwall carbon nanotube aggregate exhibiting this significant cell response was observed to be identical in structure to multiwall carbon nanotube aggregates demonstrated to be ubiquitous in the environment, and especially in indoor environments, where natural gas or propane cooking stoves exist. Correspondingly, preliminary epidemiological data, although sparse, indicate a correlation between asthma incidence or classification, and exposure to gas stoves. These results suggest a number of novel epidemiological and etiological avenues for asthma triggers and related respiratory or other environmental health effects, especially since indoor number concentrations for multiwall carbon nanotube aggregates is at least 10 times the outdoor concentration, and virtually all gas combustion processes are variously effective sources. These results also raise concerns for manufactured carbon nanotube aggregates, and related fullerene nanoparticles. PMID- 16705800 TI - Health risk assessment of cyanobacterial (blue-green algal) toxins in drinking water. AB - Cyanobacterial toxins have caused human poisoning in the Americas, Europe and Australia. There is accumulating evidence that they are present in treated drinking water supplies when cyanobacterial blooms occur in source waters. With increased population pressure and depleted groundwater reserves, surface water is becoming more used as a raw water source, both from rivers and lakes/reservoirs. Additional nutrients in water which arise from sewage discharge, agricultural run off or storm water result in overabundance of cyanobacteria, described as a 'water bloom'. The majority of cyanobacterial water-blooms are of toxic species, producing a diversity of toxins. The most important toxins presenting a risk to the human population are the neurotoxic alkaloids (anatoxins and paralytic shellfish poisons), the cyclic peptide hepatotoxins (microcystins) and the cytotoxic alkaloids (cylindrospermopsins). At the present time the only cyanobacteral toxin family that have been internationally assessed for health risk by the WHO are the microcystins, which cause acute liver injury and are active tumour promoters. Based on sub-chronic studies in rodents and pigs, a provisional Guideline Level for drinking water of 1 microg/L of microcystin-LR has been determined. This has been adopted in legislation in countries in Europe, South America and Australasia. This may be revised in the light of future teratogenicity, reproductive toxicity and carcinogenicity studies. The other cyanobacterial toxin which has been proposed for detailed health risk assessment is cylindrospermopsin, a cytotoxic compound which has marked genotoxicity, probable mutagenicity, and is a potential carcinogen. This toxin has caused human poisoning from drinking water, and occurs in water supplies in the USA, Europe, Asia, Australia and South America. An initial health risk assessment is presented with a proposed drinking water Guideline Level of 1 microg/L. There is a need for both increased monitoring data for toxins in drinking water and epidemiological studies on adverse health effects in exposed populations to clarify the extent of the health risk. PMID- 16705801 TI - Analysis of gene regulation in rabbit corneal epithelial cells induced by ultraviolet radiation. AB - Ultraviolet (UV)-induced cataracts are becoming a major environmental health concern because of the possible decrease in the stratospheric ozone layer. Experiments were designed to isolate gene(s) affected by UV irradiation in rabbit cornea tissues using fluorescent differential display-reverse transcription polymerase chain reaction (FDDRT-PCR). The epithelial cells were grown in standard medium for 2 or 4 hours post treatment. Cornea epithelial cells were irradiated with UVB for 20 minutes. RNA was extracted and amplified by reverse transcriptase-polymerase chain reaction using poly A+ specific anchoring primers and random arbitrary primers. Polyacrylamide gel electrophoresis revealed several differentially expressed genes in untreated versus UV irradiated cells. Complimentary DNA (cDNA) fragments resulting from fluorescent differentially expressed mRNAs were eluted from the gel and re-amplified. The re-amplified PCR products were cloned directly into the PCR-TRAP cloning system. These data showed that FDDRT-PCR is a useful technique to elucidate UV-regulated gene expressions. Future experiments will involve sequence analysis of cloned inserts. The identification of these genes through sequence analysis could lead to a better understanding of cataract formation via DNA damage and mechanisms of prevention. PMID- 16705802 TI - Studies on the development of potential biomarkers for rapid assessment of copper toxicity to freshwater fish using Esomus danricus as model. AB - Living in an environment that has been altered considerably by anthropogenic activities, fish are often exposed to a multitude of stressors including heavy metals. Copper ions are quite toxic to fish when concentrations are increased in environmental exposures often resulting in physiological, histological, biochemical and enzymatic alterations in fish, which have a great potential to serve as biomarkers. Esomus danricus was chosen as model in the present study and the metabolic rate, gill morphology, total glycogen, total protein, superoxide dismutase and catalase were critically evaluated. The 96h LC50 value was found to be 5.5mg/L (Cu as 1.402 mg/L). Fish groups were separately exposed to lethal (5.5 mg/L) and sub lethal concentrations (0.55 mg/L) of copper sulphate over a period of 96h to examine the subtle effects caused at various functional levels. Controls were also maintained simultaneously. Significant decrease in the metabolic rate (p<0.001) of the fish was observed in both the concentrations studied. Studies employing Automated Video Tracking System revealed gross changes in the architecture of gill morphology like loss, fusion, clubbing of secondary gill lamellae, and detachment of gill rakers following softening of gill shaft in fish under lethal exposures indicating reduced respiratory surface area. Biochemical profiles like total glycogen and total protein in gills and muscle of fish exposed to 5.5 mg/L showed appreciable decrease (p<0.05 to 0.001) from control. Significant inhibition of superoxide dismutase (60.83%), catalase (71.57%) from control was observed in fish exposed to 5.5 mg/L at the end of 96h exposure only. Interestingly, in fish exposed to 0.55 mg/L enzyme activity is not affected except for catalase. Toxic responses evaluated at various functional levels are more pronounced in fish exposed to 5.5 mg/L and these can serve as potential biomarkers for rapid assessment of acute copper toxicity in environmental biomonitoring. PMID- 16705803 TI - Metabolic activation of the tumorigenic pyrrolizidine alkaloid, retrorsine, leading to DNA adduct formation in vivo. AB - Pyrrolizidine alkaloids are naturally occurring genotoxic chemicals produced by a large number of plants. The high toxicity of many pyrrolizidine alkaloids has caused considerable loss of free-ranging livestock due to liver and pulmonary lesions. Chronic exposure of toxic pyrrolizidine alkaloids to laboratory animals induces cancer. This investigation studies the metabolic activation of retrorsine, a representative naturally occurring tumorigenic pyrrolizidine alkaloid, and shows that a genotoxic mechanism is correlated to the tumorigenicity of retrorsine. Metabolism of retrorsine by liver microsomes of F344 female rats produced two metabolites, 6, 7-dihydro-7-hydroxy-1-hydroxymethyl 5H-pyrrolizine (DHP), at a rate of 4.8 +/- 0.1 nmol/mg/min, and retrorsine-N oxide, at a rate of 17.6 +/- 0.5 nmol/mg/min. Metabolism was enhanced 1.7-fold by using liver microsomes prepared from dexamethasone-treated rats. DHP formation was inhibited 77% and retrorsine N-oxide formation was inhibited 29% by troleandomycin, a P450 3A enzyme inhibitor. Metabolism of retrorsine with lung, kidney, and spleen microsomes from dexamethasone-treated rats also generated DHP and the N-oxide derivative. When rat liver microsomal metabolism of retrorsine occurred in the presence of calf thymus DNA, a set of DHP-derived DNA adducts was formed; these adducts were detected and quantified by using a previously developed 32P-postlabeling/HPLC method. These same DNA adducts were also found in liver DNA of rats gavaged with retrorsine. Since DHP-derived DNA adducts are suggested to be potential biomarkers of riddelliine-induced tumorigenicity, our results indicate that (i) similar to the metabolic activation of riddelliine, the mechanism of retrorsine-induced carcinogenicity in rats is also through a genotoxic mechanism involving DHP; and (ii) the set of DHP-derived DNA adducts found in liver DNA of rats gavaged with retrorsine or riddelliine can serve as biomarkers for the tumorigenicity induced by retronecine-type pyrrolizidine alkaloids. PMID- 16705804 TI - Serum acetyl cholinesterase as a biomarker of arsenic induced neurotoxicity in sprague-dawley rats. AB - Arsenic is an environmental toxicant, and one of the major mechanisms by which it exerts its toxic effect is through an impairment of cellular respiration by inhibition of various mitochondrial enzymes, and the uncoupling of oxidative phosphorylation. Most toxicity of arsenic results from its ability to interact with sulfhydryl groups of proteins and enzymes, and to substitute phosphorus in a variety of biochemical reactions. Most toxicity of arsenic results from its ability to interact with sulfhydryl groups of proteins and enzymes, and to substitute phosphorus in a variety of biochemical reactions. Recent studies have pointed out that arsenic toxicity is associated with the formation of reactive oxygen species, which may cause severe injury/damage to the nervous system. The main objective of this study was to conduct biochemical analysis to determine the effect of arsenic trioxide on the activity of acetyl cholinesterase; a critical important nervous system enzyme that hydrolyzes the neurotransmitter acetylcholine. Four groups of six male rats each weighing an average 60 +/- 2 g were used in this study. Arsenic trioxide was intraperitoneally administered to the rats at the doses of 5, 10, 15, 20mg/kg body weight (BW), one dose per 24 hour given for five days. A control group was also made of 6 animals injected with distilled water without chemical. Following anaesthesia, blood specimens were immediately collected using heparinized syringes, and acetyl cholinesterase detection and quantification were performed in serum samples by spectrophotometry. Arsenic trioxide exposure significantly decreased the activity of cholinesterase in the Sprague-Dawley rats. Acetyl cholinesterase activities of 6895 +/- 822, 5697 +/- 468, 5069 +/- 624, 4054 +/- 980, and 3158 +/- 648 U/L were recorded for 0, 5, 10, 15, and 20 mg/kg, respectively; indicating a gradual decrease in acetyl cholinesterase activity with increasing doses of arsenic. These findings indicate that acetyl cholinesterase is a candidate biomarker for arsenic-induced neurotoxicity in Sprague-Dawley rats. PMID- 16705805 TI - Prevalence of selected bacterial infections associated with the use of animal waste in Louisiana. AB - Human health is a major concern when considering the disposal of large quantities of animal waste. Health concerns could arise from exposure to pathogens and excess nitrogen associated with this form of pollution. The objective was to collect and analyze health data related to selected bacterial infections associated with the use of animal waste in Louisiana. An analysis of adverse health effects has been conducted based on the incidence/prevalence rates of campylobacteriosis, E. coli O157:H7 infection, salmonellosis and shigellosis. The number of reported cases increased during the summer months. Analysis of health data showed that reported disease cases of E. coli O157:H7 were highest among Caucasian infants in the 0-4 year old age category and in Caucasian children in the 5-9 year old age category. Fatalities resulting from salmonellosis are low and increases sharply with age. The number of reported cases of shigellosis was found to be higher in African American males and females than in Caucasians. The high rate of identification in the younger population may result from the prompt seeking of medical care, as well as the frequent ordering of stool examination when symptoms become evident among this group of the population. The association with increasing age and fatality due to salmonellosis could be attributed to declining health and weaker immune systems often found in the older population. It is concluded that both animal waste and non-point source pollution may have a significant impact on human health. PMID- 16705806 TI - Comparative assessment of the physico-chemical and bacteriological qualities of selected streams in Louisiana. AB - The objective of this research was to compare the chemical/physical parameters and bacterial qualities of selected surface water streams in Louisiana, including a natural stream (control) and an animal waste related stream. Samples were collected and analyzed for fecal coliforms. Fecal coliforms isolated from these samples were identified to the species level. Chemical analysis was performed following standard test protocols (LaMotte 2002). An analysis of biological oxygen demand (BOD), chemical oxygen demand (COD), total organic carbon (TOC), total dissolved solids (TDS), conductivity, pH, temperature, ammonia nitrogen, nitrate nitrogen, iron, copper, phosphate, potassium, sulfate, turbidity, zinc and bacterial levels was performed following standard test protocols as presented in Standard Methods for the Examination of Water and Wastewater [9]. Results of the comparisons of the various surface water streams showed that phosphate levels, according to Mitchell and Stapp, were considered good for Lake Claiborne (control) and Bayou Dorcheat. The levels were found to be .001 mg/L and .007 mg/L respectively. Other streams associated with animal waste, had higher phosphate levels of 2.07 mg/L and 2.78 mg/L, respectively. Conductivity and total dissolved solids (TDS) levels were the lowest in Lake Claiborne and highest in the Hill Farm Research Station stream. It can be concluded from the data that some bacterial levels and various nutrient levels can be affected in water resources due to non-point source pollution. Many of these levels will remain unaffected. PMID- 16705807 TI - Cytogenetic effects of 1,1-dichloroethane in mice bone marrow cells. AB - The major concern for the halogenated compounds is their widespread distribution, in addition to occupational exposures. Several chlorinated alkanes and alkenes were found to induce toxic effects. In this study, we investigated the genotoxic potential of 1,1-dichloroethane in the bone marrow cells obtained from Swiss Webster mice, using chromosomal aberrations (CA), mitotic index (MI), and micronuclei (MN) formation as toxicological endpoints. Five groups of three male mice each, weighing an average of 24 +/- 2 g, were injected intraperitoneally, once with doses of 100, 200, 300, 400, 500 mg/kg body weight (BW) of 1,1 dichloroethane dissolved in ethanol. A control group was also made of three animals injected with ethanol (1%) without the chemical. All animals were sacrificed 24 hours after the treatment. Chromosome and micronuclei preparations were obtained from bone marrow cells following standard protocols. Chromatid and chromosome aberrations were investigated in 100 metaphase cells per animal and percent micronuclei frequencies were investigated in 1,000 metaphase cells per animal. 1,1-dichloroethane exposures significantly increased the number of chromosomal aberrations and the frequency of micronucleated cells in the bone marrow cells of Swiss-Webster mice. Percent chromosomal aberrations of 2.67 +/- 0.577, 7.66 +/- 2.89, 8.33 +/- 2.08, 14.67 +/- 2.51, 20.3 +/- 3.21, 28 +/- 3.61; mitotic index of 9.4%, 7.9%, 6.2%, 4.3%, 3.0%, 2.6% and micronuclei frequencies of 3.33 +/- 0.7, 7.33 +/- 0.9, 8.00 +/- 1.0, 11.67 +/- 1.2, 15.33 +/- 0.7, 18.00 +/- 1.7 were recorded for the control, 100, 200, 300, 400, and 500 mg/kg BW respectively; indicating a gradual increase in number of chromosomal aberrations and micronuclei formation, with increasing dose of 1,1,-dichloroethane. Our results indicate that 1,1-dichloroethane has a genotoxic potential as measured by the bone marrow CA and MN tests in Swiss-Webster mice. PMID- 16705809 TI - Photochemical reaction of 7,12-dimethylbenz[a]anthracene (DMBA) and formation of DNA covalent adducts. AB - DMBA, 7,12-dimethylbenz[a]anthracene, is a widely studied polycyclic aromatic hydrocarbon that has long been recognized as a probable human carcinogen. It has been found that DMBA is phototoxic in bacteria as well as in animal or human cells and photomutagenic in Salmonella typhimurium strain TA102. This article tempts to explain the photochemistry and photomutagenicity mechanism. Light irradiation converts DMBA into several photoproducts including benz[a]anthracene 7,12-dione, 7-hydroxy-12-keto-7-methylbenz[a]anthracene, 7,12-epidioxy-7,12 dihydro-DMBA, 7-hydroxymethyl-12-methylbenz[a]anthracene and 12-hydroxymethyl-7 methylbenz[a]anthracene. Structures of these photoproducts have been identified by either comparison with authentic samples or by NMR/MS. At least four other photoproducts need to be assigned. Photo-irradiation of DMBA in the presence of calf thymus DNA was similarly conducted and light-induced DMBA-DNA adducts were analyzed by 32P-postlabeling/TLC, which indicates that multiple DNA adducts were formed. This indicates that formation of DNA adducts might be the source of photomutagenicity of DMBA. Metabolites obtained from the metabolism of DMBA by rat liver microsomes were reacted with calf thymus DNA and the resulting DNA adducts were analyzed by 32P-postlabeling/TLC under identical conditions. Comparison of the DNA adduct profiles indicates that the DNA adducts formed from photo-irradiation are different from the DNA adducts formed due to the reaction of DMBA metabolites with DNA. These results suggest that photo-irradiation of DMBA can lead to genotoxicity through activation pathways different from those by microsomal metabolism of DMBA. PMID- 16705808 TI - Diesel exhaust particles induce the over expression of tumor necrosis factor alpha (TNF-alpha) gene in alveolar macrophages and failed to induce apoptosis through activation of nuclear factor-kappaB (NF-kappaB). AB - Exposure to particulate matter (PM2.5-10), including diesel exhaust particles (DEP) has been reported to induce lung injury and exacerbation of asthma and chronic obstructive pulmonary disease. Alveolar macrophages play a major role in the lung's response to inhaled particles and therefore, are a primary target for PM2.5-10 effect. The molecular and cellular events underlying DEP-induced toxicity in the lung, however, remain unclear. To determine the effect of DEP on alveolar macrophages, RAW 264.7 cells were grown in RPMI 1640 with supplements until confluency. RAW 264.7 cultures were exposed to Hank's buffered saline solution (vehicle), vehicle containing an NF-kappaB inhibitor, BAY11-7082 (25 microM with 11/2 hr pre-incubation), or vehicle containing DEP (250 microg/ml) in the presence or absence of BAY11-7082 (25 microM with 11/2 hr pre-incubation) for 4 hr and TNF-alpha release was determined by enzyme-linked immunosorbent assay and confirmed by western blots. RAW 264.7 apoptotic response was determined by DNA fragmentation assays. U937 cells treated with campothecin (4 microg/ml x 3 hr), an apoptosis-inducing agent, were used as positive control. We report that exposure to the carbonaceous core of DEP induces significant release of TNF-alpha in a concentration-dependent fashion (31 +/- 4 pg/ml, n = 4, p = 0.08; 162 +/- 23 pg/ml, n = 4, p < 0.05; 313 +/- 31 pg/ml, n = 4, p < 0.05 at 25, 100, and 250 microg/ml, respectively). DEP exposure, however, failed to induce any apoptotic response in RAW 264.7 cells. Moreover, inhibition of NF-kappaB binding activity has resulted in DEP-induced apoptotic response in alveolar macrophages, as demonstrated by the NF-kappaB inhibitor, BAY11-7082 studies. The results of the present study indicate that DEP induce the release of TNF-alpha in alveolar macrophages, a primary target for inhaled particles effect. DEP-induced TNF-alpha gene expression is regulated at the transcriptional level by NF-kappaB. Furthermore, DEP-induced increase in NF-kappaB-DNA binding activity appears to protect against apoptosis. PMID- 16705810 TI - Cytokines and adhesion molecules expression in the brain in human cerebral malaria. AB - Although the role of systemic proinflammatory cytokines, IL-1beta and TNF-alpha, and their up-regulation of adhesion molecules, ICAM-1, VCAM-1 and E-Selectin, in the pathogenesis of cerebral malaria (CM) is well established, the role of local cytokine release remain unclear. Immunohistochemistry (IHC) was used to compare the expression of ICAM-1, VCAM-1, E-Selectin, IL-1beta, TNF-a and TGF-beta at light microscopic level in cerebral, cerebellar and brainstem postmortem cryostat sections from 10 CM, 5 severe malarial anemia (SMA), 1 purulent bacterial meningitis (PBM), 2 non-central nervous system infections (NCNSI) and 3 non infections (NI) deaths in Ghanaian children. Fatal malaria and Salmonella sepsis showed significantly higher vascular expression of all 3 adhesion molecules, with highly significant co-localization with sequestration in the malaria cases. However, there was negligible difference between CM and SMA. TGF-beta showed intravascular and perivascular distribution in all cases, but expression was most intense in the PBM case and CM group. TNF-alpha and IL-1beta showed prominent brain parenchymal staining, in addition to intravascular and perivascular staining, in only the PBM case and CM group. The maximal expression of all 6 antigens studied was in the cerebellar sections of the malaria cases. Endothelial activation is a feature of fatal malaria and Salmonella sepsis, with adhesion molecule expression being highly correlated with sequestration. IL-1beta and TNF alpha are upregulated in only cases with neurodegenerative lesions, whilst TGF beta is present in all cases. Both cytokines and adhesion molecules were maximally upregulated in the cerebellar sections of the malaria cases. PMID- 16705811 TI - Effect of co-existing biologically relevant molecules and ions on DNA photocleavage caused by pyrene and its derivatives. AB - Inorganic ions, coenzymes, amino acids, and saccharides could co-exist with toxic environmental chemicals, such as polycyclic aromatic hydrocarbons (PAHs), in the cell. The presence of these co-existing chemicals can modulate the toxicity of the PAHs. One of the genotoxic effects by PAHs is light-induced cleavage, or photocleavage, of DNA. The effect of inorganic ions I-, Na+, Ca2+, Mg2+, Fe3+, Mn2+, Cu2+, and Zn2+ and biological molecules riboflavin, histidine, mannitol, nicotinamide adenine dinucleotide (NAD), glutathione, and glutamic acid on the DNA photocleavage by pyrene, 1-hydroxypyrene (1-HP), and 1-aminopyrene (1-AP), is studied. The non-transition metal ions Na+, Ca2+, and Mg2+, usually have very little inhibitory effects, while the transition metal ions Fe3+, Cu2+, and Zn2+ enhance, Mn2+ inhibits the DNA photocleavage. The effect by biological molecules is complex, depending on the photochemical reaction mechanisms of the compounds tested (1-AP, 1-HP and pyrene) and on the chemical nature of the added biological molecules. Riboflavin, histidine, and mannitol enhance DNA photocleavage by all three compounds, except that mannitol has no effect on the photocleavage of DNA by pyrene. Glutathione inhibits the DNA photocleavage by 1-AP and 1-HP, but has no effect on that by pyrene. NAD enhances the DNA photocleavage by 1-AP, but has no effect on that by 1-HP and pyrene. Glutamic acid enhances the DNA photocleavage by 1-AP and pyrene, but inhibits that by 1-HP. These results show that the co-existing chemicals may have a profound effect on the toxicity of PAHs, or possibly on the toxicity of many other chemicals. Therefore, if one studies the toxic effects of PAHs or other toxic chemicals, the effect of the co existing chemicals or ions needs to be considered. PMID- 16705812 TI - Photodecomposition and phototoxicity of natural retinoids. AB - Sunlight is a known human carcinogen. Many cosmetics contain retinoid-based compounds, such as retinyl palmitate (RP), either to protect the skin or to stimulate skin responses that will correct skin damaged by sunlight. However, little is known about the photodecomposition of some retinoids and the toxicity of these retinoids and their sunlight-induced photodecomposition products on skin. Thus, studies are required to test whether topical application of retinoids enhances the phototoxicity and photocarcinogenicity of sunlight and UV light. Mechanistic studies are needed to provide insight into the disposition of retinoids in vitro and on the skin, and to test thoroughly whether genotoxic damage by UV-induced radicals may participate in any toxicity of topically applied retinoids in the presence of UV light. This paper reports the update information and our experimental results on photostability, photoreactions, and phototoxicity of the natural retinoids including retinol (ROH), retinal, retinoid acid (RA), retinyl acetate, and RP (Figure 1). PMID- 16705813 TI - Effect of light irradiation and sex hormones on jurkat T cells: 17beta-estradiol but not testosterone enhances UVA-induced cytotoxicity in Jurkat lymphocytes. AB - In Eastern cultures, such as India, it is traditionally recommended that women but not men cover their heads while working in the scorching sun. The purpose of this pilot study was to determine whether there was any scientific basis for this cultural tradition. We examined the differential cytotoxic effects of ultraviolet A light (UVA) on an established T cell line treated with female and male sex hormones. CD4+ Jurkat T cells were plated in 96 well plates at 2 x 106 cells/ml and treated with 17beta-estradiol (EST) or testosterone (TE). These cells were irradiated by UVA light with an irradiance of 170 J/cm2 for 15min at a distance of 6 cm from the surface of the 96-well plate. Controls included cells not treated with hormones or UVA. The effects of EST and TE were investigated between 1 and 20 ng/mL. Cytotoxicity by fluorescein-diacetate staining and COMET assay generating single strand DNA cleavage, tail length and tail moment measurements were examined. The effect of estrogen (5ng/mL) on apoptosis and its mediators was further studied using DNA laddering and western blotting for bcl-2 and p53. We found that EST alone, without UVA, enhanced Jurkat T cell survival. However, EST exhibited a dose-related cytotoxicity in the presence of UVA; up to 28% at 20 ng/ml. TE did not alter UVA-induced cytotoxicity. Since TE did not alter cell viability in the presence of UVA further damaging studies were not performed. COMET assay demonstrated the harmful effects of EST in the presence of UVA while EST without UVA. had no significant effect on the nuclear damage. Apoptosis was not present as indicated by the absence of DNA laddering on agarose gel electrophoresis at 5ng/ml EST or TE +/- UVA. Western blot showed that estrogen down regulated bcl-2 independently of UVA radiation while p53 was down regulated in the presence of UVA treatment. EST and TE have differential effects on UVA induced cytotoxicity in Jurkat T-lymphocyte which suggested that women may be more susceptible to the harmful effects of solar irradiation than men. PMID- 16705814 TI - Assessment of lead exposure risk in locksmiths. AB - Exposure to lead has been well recognized in a number of work environments, but little is known about lead exposure associated with machining brass keys containing lead. The brass that is widely used for key manufacturing usually contains 1.5% - 2.5 % of lead. Six (6) licensed locksmiths and 6 case-matched controls successfully completed the pilot study to assess the prevalence of increased body lead burden of professional locksmiths. We measured both Blood Lead (atomic absorption spectrometry), bone-lead (KXRF) and had each subject complete a health and lead exposure risk questionnaire. One locksmith had not cut keys during the past two years, therefore this subject and case-matched control was excluded from the blood lead analysis only. The average blood-lead concentration (+/-SEM) for the 5 paired subjects was 3.1 (+/- 0.4) microg/dL and 2.2 (+/- 0.3) microg/dL for controls. Bone measurements, including all 6 paired subjects, showed tibia lead concentration (+/-SEM) for locksmiths and controls was 27.8 (+/- 2.3) microg/g and 13.7 (+/- 3.3) microg/g, respectively; average calcaneus lead concentration for locksmiths and controls was 31.9 (+/- 3.7) microg/g and 22.6 (+/- 4.1) microg/g, respectively: The t-test shows a significantly higher tibia lead (p<0.05) and blood lead (p<0.05) for locksmiths than for their matched controls, but no significant difference for calcaneus lead (p>0.10). Given that the mean tibia bone lead concentration was 13.1 microg/g higher in locksmiths than in their matched controls, this average difference in the two groups would translate to an OR of increased hypertension in locksmiths of between 1.1 and 2.3, based on the published literature. Even with the very small number of subjects participating in this pilot study, we were able to demonstrate that locksmiths had significantly higher current exposure to lead (blood lead concentration) and significantly higher past exposure to lead (tibia lead concentration) than their age, sex and ethnically matched controls. Additional research is needed to fully identify the prevalence and associated risk factors for occupational exposure of lead in this previously understudied profession. PMID- 16705815 TI - Synthesis and structure-activity correlation studies of metal complexes of alpha N-heterocyclic carboxaldehyde thiosemicarbazones in Shewanella oneidensis. AB - This investigation involved the synthesis of metal complexes to test the hypothesis that structural changesand metal coordination in pyridine thiosemicarbazones affect cell growth and cell proliferation in vitro. Thiosemicarbazones are well known to possess antitumor, antiviral, antibacterial, antimalarial, and other activities. Extensive research has been carried out on aliphatic, aromatic, heterocyclic and other types of thiosemicarbazones and their metal complexes. Due to the pronounced reactivity exhibited by metal complexes of heterocyclic thiosemicarbazones, synthesis and structural characterization of di 2-pyridylketone 4N-phenyl thiosemicarbazone and diphenyl tin (Sn) and platinum (Pt) complexes were undertaken. Shewanella oneidensis MR-1, a metal ion-reducing bacterium, was used as a model organism to explore the biological activity under aerobic conditions. A comparision of the cytotoxic potential of selected ligand and metal-complex thiosemicarbazones on cell growth in wild type MR-1 and mutant DSP-010 Shewanella oneidensis strains at various concentrations (0, 5, 10, 15, 20 or 25 ppm) was performed. The wild type (MR-1) grown in the presence of increasing concentrations of Sn- thiosemicarbazone complexes was comparatively more sensitive (mean cell number = 4.8 X 10(8) +/- 4.3 X 10(7) SD) than the DSP 010, a spontaneous rifampicillin derivative of the parent strain (mean cell number = 5.6 x 10(8) +/- 6.4 X 10(7) SD) under comparable aerobic conditions (p = 0.0004). No differences were observed in the sensitivity of the wild and mutant types when exposed to various concentrations of diphenyl Pt- thiosemicarbazone complex (p = 0.425) or the thiosemicarbazone ligand (p = 0.313). Growth of MR-1 in the presence of diphenyl Sn-thiosemicarbazone was significantly different among treatment groups (p = 0.012). MR-1 cell numbers were significantly higher at 5ppm than at 10 to 20ppm (p = 0.05). The mean number of DSP-010 variant strain cells also differed among diphenyl Sn- thiosemicarbazone complex treated groups (p = 0.051). In general, there was an increasing trend in the number of cells from about 5.0 X 10(8) cells (methanol control group) to about 6.0 X 10(8) cells (25ppm). The number of cells in methanol control group was significantly lower than cell numbers at 20ppm and 25ppm (p = 0.05), and numbers at 5ppm treatment were lower than at 20 and 25ppm (p = 0.05). Furthermore, a marginally significant difference in the number of MR-1 cells was observed among diphenyl Pt- thiosemicarbazone complex treatment groups (p = 0.077), and an increasing trend in the number of cells was noted from approximately 5.0 X 10(8) cells (methanol control group) to approximately 5.8 X 10(8) cells (20ppm). In contrast, the DSP 010 variant strain showed no significant differences in cell numbers when treated with various concentrations of diphenyl Pt- thiosemicarbazone complex (p = 0.251). Differences in response to Sn- metal complex between MR-1 and DSP-010 growing cultures indicate that biological activity to thiosemicarbazone metal complexes may be strain specific. PMID- 16705816 TI - Application of oligonucleotide microarrays for bacterial source tracking of environmental Enterococcus sp. isolates. AB - In an effort towards adapting new and defensible methods for assessing and managing the risk posed by microbial pollution, we evaluated the utility of oligonucleotide microarrays for bacterial source tracking (BST) of environmental Enterococcus sp. isolates derived from various host sources. Current bacterial source tracking approaches rely on various phenotypic and genotypic methods to identify sources of bacterial contamination resulting from point or non-point pollution. For this study Enterococcus sp. isolates originating from deer, bovine, gull, and human sources were examined using microarrays. Isolates were subjected to Box PCR amplification and the resulting amplification products labeled with Cy5. Fluorescent-labeled templates were hybridized to in-house constructed nonamer oligonucleotide microarrays consisting of 198 probes. Microarray hybridization profiles were obtained using the ArrayPro image analysis software. Principal Components Analysis (PCA) and Hierarchical Cluster Analysis (HCA) were compared for their ability to visually cluster microarray hybridization profiles based on the environmental source from which the Enterococcus sp. isolates originated. The PCA was visually superior at separating origin-specific clusters, even for as few as 3 factors. A Soft Independent Modeling (SIM) classification confirmed the PCA, resulting in zero misclassifications using 5 factors for each class. The implication of these results for the application of random oligonucleotide microarrays for BST is that, given the reproducibility issues, factor-based variable selection such as in PCA and SIM greatly outperforms dendrogram-based similarity measures such as in HCA and K-Nearest Neighbor KNN. PMID- 16705817 TI - How much should we involve genetic and environmental factors in the risk assessment of mycotoxins in humans? AB - Despite consented efforts in prevention, mycotoxins remain a problem of human health concern in several parts of the world including developed countries. Within the same range of toxins concentrations in the blood some people develop a disease while others do not. Could this inequality in front of mycotoxins effects be explained by environment factors and/or genetic predisposition? Among recent advances in environmental health research Correlation between chronic diseases and mycotoxins in humans deserves attention through several questions: Are genetic factors involved in disease causation of mycotoxins? How much are these factors currently taken into account for mycotoxins risk assessment and how much should we involve them? Answers are still to come. Genetic and environment factors deserve therefore more attention when dealing with regulatory limits, since among the general population, those who are at risk and will develop specific diseases are likely those bearing genetic predispositions. We have addressed these questions for the specific case of ochratoxin A in humans by investigating in Tunisia, county of Jelma, in four rural families forming a household of 21 persons all exposed to ochratoxin A in diet. Our results confirm that ochratoxin A induces chronic tubular nephropathy in humans and mainly point at those having the HLA haplotype A3, B27/35, DR7 to be more sensitive to the disease for quantitatively similar or lower exposure. Persons with such haplotype were found to bear chronic interstitial nephropathy with tubular karyomegalic cells while others were apparently healthy. Godin et al. (1996) in France have also found in sibling (a sister and her brother from urban area) that have similar HLA haplotype B35-patern, OTA-related renal tubulopathy with mild proteinuria including beta2-microglobulinuria. Several mechanisms are discussed that could be put ahead to explain how the HLA haplotype could lead to tubular cells lyses and renal failure. In the mean time it is urgent to search for mass screening biomarkers for mycotoxins in humans and related genetic factors to set up more appropriate regulation. PMID- 16705818 TI - Conceptual modeling for adaptive environmental assessment and management in the Barycz Valley, lower Silesia, Poland. AB - The complexity of interactions in socio-ecological systems makes it very difficult to plan and implement policies successfully. Traditional environmental management and assessment techniques produce unsatisfactory results because they often ignore facets of system structure that underlie complexity: delays, feedbacks, and non-linearities. Assuming that causes are linked in a linear chain, they concentrate on technological developments ("hard path") as the only solutions to environmental problems. Adaptive Management is recognized as a promising alternative approach directly addressing links between social and ecological systems and involving stakeholders in the analysis and decision process. This "soft path" requires special tools to facilitate collaboration between "experts" and stakeholders in analyzing complex situations and prioritizing policies and actions. We have applied conceptual modeling to increase communication, understanding and commitment in the project of seven NGOs "Sustainable Regional Development in the Odra Catchment". The main goal was to help our NGO partners to facilitate their efforts related to developing sustainable policies and practices to respond to large-scale challenges (EU accession, global changes in climate and economy) to their natural, economic and socio-cultural heritages. Among the variety of sustainability issues explored by these NGOs, two (extensive agricultural practices and "green" local products) were examined by using Adaptive Management (AM) as a framework that would link analysis, discussion, research, actions and monitoring. Within the AM framework the project coordinators used tools of systems analysis (Mental Model Mapping) to facilitate discussions in which NGO professionals and local stakeholders could graphically diagram and study their understanding of what factors interacted and how they affect the region's sustainability. These discussions produced larger scale Regional Sustainability Models as well as more detailed sub-models of particular factors, processes, and feedback loops that appear critical to a sustainable future. The Regional Sustainability Model was used to identify a subset of key interacting factors (variables). For each variable, several sustainability indicators were suggested. The growing understanding and acceptance of the AM framework and systems analysis created a momentum both locally and within the region, which makes continued successful use of these indicators quite likely. In contrast to expert-driven projects that inject outside knowledge into a local context, this project established a broad basis for stakeholder-driven discussion that is articulated into goals, objectives, conceptual models, and indicators. The ability to learn and adapt in the AM framework increases the capacity to innovate and find policies and practices that enhance resilience and sustainability in a world in transition. PMID- 16705820 TI - Toxic aluminium and heavy metals in groundwater of middle Russia: health risk assessment. AB - Two approaches are distinguished in modern ecological monitoring. The first one is physicochemical analysis of environmental objects with respect to maximum allowable concentrations (MACs) of chemical substances, which is performed by standards methods in accordance with state regulations. The second approach (biological monitoring) is based on the methodology of biotesting and bio indication. The task of this work is to create biotests for estimation of Al and other metals toxicity in ground water and to compare these results with physicochemical analysis dates. Risk assessment for heavy metals contaminated groundwater was also performed. Risk assessment was performed accordingly EPA US recommendation and gave results about 90 per 100000 citizens for Al and 402 per 100000 for mixture of different heavy metals. For comparison: risk for earth background radiation for Middle Russia is (Individual dose 1 millisivert per year) consist 5 per 100000 people. It was shown that groundwater consist HCO3- ions (360 mg/l), sometimes Al compounds 0.21-0.65 mg/l (MAC for Al is 0.5 mg/l for Russia). Other groundwater contain Hg - 0.004 mg/l (MAC - 0.0005 mg/l); Cr - 0.072 mg/l (MAC - 0.05 mg/l); As - less than 0.03 mg/l (MAC - 0.05 mg/l). We developed plant biotest for estimation of groundwater quality with barley roots, tradescatia and others. Some biotests parameters correlate with HCO3-, Cl-, SO(4)2- and metal ions content positively, for another biotest this correlation is strongly negative. The quality of groundwater near Obninsk and in Kaluga Region is very different but hasnit been changed since the year 1998. PMID- 16705819 TI - Assessment of arsenic contamination of groundwater and health problems in Bangladesh. AB - Excessive amounts of arsenic (As) in the groundwater in Bangladesh and neighboring states in India are a major public health problem. About 30% of the private wells in Bangladesh exhibit high concentrations of arsenic. Over half the country, 269 out of 464 administrative units, is affected. Similar problems exist in many other parts of the world, including the Unites States. This paper presents an assessment of the health hazards caused by arsenic contamination in the drinking water in Bangladesh. Four competing hypotheses, each addressing the sources, reaction mechanisms, pathways, and sinks of arsenic in groundwater, were analyzed in the context of the geologic history and land-use practices in the Bengal Basin. None of the hypotheses alone can explain the observed variability in arsenic concentration in time and space; each appears to have some validity on a local scale. Thus, it is likely that several biogeochemical processes are active among the region's various geologic environments, and that each contributes to the mobilization and release of arsenic. Additional research efforts will be needed to understand the relationships between underlying biogeochemical factors and the mechanisms for arsenic release in various geologic settings. PMID- 16705822 TI - Lead accumulation by tall fescue (Festuca arundinacea Schreb.) grown on a lead contaminated soil. AB - Phytoextraction is gaining acceptance as a cost-effective and environmentally friendly phytoremediation strategy for reducing toxic metal levels from contaminated soils. Cognizant of the potential of this phytoremediation technique as an alternative to expensive engineering-based remediation technologies, experiments were conducted to evaluate the suitability of some plants as phytoextraction species. From one of our preliminary studies, we found that tall fescue (Festuca arundinacea Schreb. cv. Spirit) can tolerate and accumulate significant amounts of lead (Pb) in its shoots when grown in Pb-amended sand. To further evaluate the suitability of tall fescue as one of the potential crop rotation species for phytoextraction, a study was conducted to determine whether the addition of ethylenediaminetetraacetic acid (EDTA) alone or in combination with acetic acid can further enhance the shoot uptake of Pb. Seeds were planted in 3.8 L plastic pots containing top soil, peat, and sand (4:2:1, v:v:v) spiked with various levels (0,1000, 2000 mg Pb/kg dry soil) of lead. At six weeks after planting, aqueous solutions (0, 5 mmol/kg dry soil) of EDTA and acetic acid (5 mmol/kg dry soil) were applied to the root zone, and all plants were harvested a week later. Results revealed that tall fescue was relatively tolerant to moderate levels of Pb as shown by non-significant differences in root and shoot biomass among treatments. An exception to this trend however, was the slight reduction in root and shoot biomass of plants exposed to the highest Pb level in combination with the two chelates. Root Pb concentration increased with increasing level of soil-applied Pb. Further increases in root Pb concentrations were attributed to chelate amendments. Translocation index, which is a measure of the partitioning of the metal to the shoots, was significantly enhanced with chelate addition especially when both EDTA and acetic acid were used. Chelate-induced increases in translocation indices correspondingly led to higher shoot Pb concentrations. PMID- 16705823 TI - GIS applications in land management: the loss of high quality land to development in Central Mississippi from 1987-2002. AB - The socio-economic trends and history of Central Mississippi reveal a major rural influence based upon a dependence on agricultural activities as part of the economic engine driving the state's economy. Yet, in the last several years, the amount of agricultural land in the counties continues to decline. Similar changes in other variables associated with agricultural land use and the continuity of farming in the state have also been changing. Indeed, under the pressure of urban growth, some farmers are forced to use less productive soils or have abandoned the agricultural business. Considering the gravity of the problem and the implications for sustainable development, public concern has increased in the state of Mississippi that urbanization and other factors may be eroding potential farmland. Given the effects of the current trends on the future capacity to produce food items, there are concerns that the growing incidence of farmland loss may also erode the basis for sustainable use of agricultural land, biodiversity and protection of the state's ecological treasures. Notwithstanding the gravity of these trends, no major effort in the literature has aimed at documenting the incidence of agricultural land loss and the linkages to urbanization in the region of Central Mississippi. What changes have taken place in the size of agricultural land within the counties and what factors are responsible for it? This paper examines the issue of farmland loss in Central Mississippi with a focus at the county level between 1987 and 2002 from a temporal-spatial perspective. In terms of methodology, the paper uses a mixed scale approach based upon the existing literature. Data were drawn from the United States Census databases of Population and Agriculture. This information is analyzed with basic descriptive statistics and GIS with particular attention to the spatial trends at the county level. Results indicate that the counties under consideration have experienced considerable change in the amount of agricultural land and other variables associated with the use of farmland, due to urbanization. With the types of changes occurring, instituting effective policies anchored in sustainability, community participation, and growth management will go a long way in addressing the situation. Other strategies for farmland protection based upon land information inventory and mapping in the region, are also recommended. The paper stands as an update of the existing literature and offers a valuable tool for decision makers within the domain of natural resources management. PMID- 16705821 TI - Evidence for alteration in chemical and physical properties of water and modulation of its biological functions by sunlight transmitted through color ranges of the visible spectrum-a novel study. AB - We investigated the changes in the properties of water when exposed to sunlight for 40 days. We hypothesize and prove that solar irradiation to water entraps electromagnetic radiation as potential energy, which becomes kinetic energy in various systems. It is postulated that photochemically-induced energy transfers, associated with individual spectral emission of visible spectrum of solar light, exert diverse influences on biological systems. Bottles of distilled water, individually wrapped in spectral-colored cellophane were exposed to sunlight and compared to an unwrapped bottle to determine chemical and physical changes as well as modifications of biological properties. Each bottle of water was named according to the color of cellophane paper with letter E (stands for exposed) as a prefix with (E-violet, E-indigo, E-blue, E-green, E-yellow, E-orange, and E red). E-control (without wrap) was exposed to polychromatic sunlight. This study addresses two main issues viz., the chemical and physical changes in E-water and its effect on biological activities. Chemical and physical composition analysis using inductively coupled plasma atomic emission spectrometry; physical conductance by a Wheatstone Bridge type conductivity meter; osmolarity by a vapor pressure osmometer; and, salt solubility profile of 10% sodium bicarbonate were determined. Furthermore, testing the effect of E-waters on human lymphocyte proliferation, mosquito larvae hatching and seed germination determined the functional role of solar radiation through specific spectrum/s of visible light on various biological processes. We found that water exposed to visible spectral emissions of sunlight had an altered elemental composition, electrical conductance, osmolarity and salt-solubility, as well as differences in bio modulatory effects. A gradual increase in leaching of Boron from E-violet to E red was noted. E-indigo showed maximal increase in electrical conductance and maximal salt solubility of sodium bicarbonate. E-blue inhibited phyto hemagglutinin-induced immune cell proliferation and mosquito larvae hatching. E orange stimulated root elongation in seed germination. We conclude that 40-day exposure of water to specific solar spectrum changes chemical and physical properties and influences on biological activity. PMID- 16705824 TI - Contribution of man-made activities to the pollution of the Tigris within Mosul area/IRAQ. AB - This paper presents an overall view of major sources that may lead to the pollution of the Tigris within Mosul city. A stretch exceeding 20kms in length is selected that represents the "sick" path of the river. Many sites along the studied stretch are likely to affect the river quality in some way or another. Samples from 40 sources sites are taken for quality analyses These sources--as huge as 400000 m3 a day--are characterized as (medium - strong) in composition. Such wastewaters with the pollutants they carry alter the river water quality rendering it unsuitable for beneficial uses. Such alterations--do leave--many negative consequences concerning human beings and aquatic life. It is found that domestic discharges are among the most important sources of pollution. Sanitary wastes are often discharged--untreated--into the Tigris. Other illegal practices such as in-house slaughtering add to the pollution as well. Industrial, tourist and institutional wastes put an additional burden on pollution of the river water quality. These wastes contain lead, chrome, and other heavy metals that may pose health risks. Wastewater treatment plants that exist in some sectors do not perform as they are expected. They need proper evaluation and rehabilitation. Eutrophication--a characteristic problem in lakes--finds an access to occur into the Tigris. This problem results from intensive use of detergents rich in nutrients (P&N compounds). In general, pollutants of different sources heavily affect the river water. Recovery and self purification of the river is estimated to occur at 40 km far from reference point. The paper concludes with the necessity of construction of a central treatment plant(s) or tackling the pollutants at their origin. The paper also stresses on importance of environmental education and awareness in order to combat pollution problems. PMID- 16705825 TI - Bioremediation of contaminated lake sediments and evaluation of maturity indicies as indicators of compost stability. AB - Land contamination is one of the widely addressed problems, which is gaining importance in many developed and developing countries. International efforts are actively envisaged to remediate contaminated sites as a response to adverse health effects. Popular conventional methodologies only transfer the phase of the contaminant involving cost intensive liabilities besides handling risk of the hazardous waste. Physico-chemical methods are effective for specific wastes, but are technically complex and lack public acceptance for land remediation. iBioremediatio ni, is one of the emerging low-cost technologies that offer the possibility to destroy various contaminants using natural biological activities. Resultant non-toxic end products due to the microbial activity and insitu applicability of this technology is gaining huge public acceptance. In the present study, composting is demonstrated as a bioremediation methodology for the stabilization of contaminated lake sediments of Hyderabad, A.P, India. Lake sediment contaminated with organics is collected from two stratums--upper (0.25 m) and lower (0.5m) to set up as Pile I (Upper) and Pile II (Lower) in the laboratory. Lime as a pretreatment to the lake sediments is carried out to ensure metal precipitation. The pretreated sediment is then mixed with organic and inorganic fertilizers like cow dung, poultry manure, urea and super phosphate as initial seeding amendments. Bulking agents like sawdust and other micronutrients are provided. Continuous monitoring of process control parameters like pH, moisture content, electrical conductivity, total volatile solids and various forms of nitrogen were carried out during the entire course of the study. The stability of the compost was evaluated by assessing maturity indices like C/N, Cw (water soluble carbon), CNw (Cw/Nw), nitrification index (NH4/NO-3), Cation Exchange Capacity (CEC), germination index, humification ratio, compost mineralization index (ash content/oxidizable carbon), sorption capacity index (CEC/oxidizable carbon). Enzyme activities of agricultural interest like urease, phosphatase, P-glucosidase, dehydrogenase and BAA-hydrolyzing protease, which are involved in the nitrogen, phosphorus and carbon cycles, were also assessed. Total content of macro and micronutrients in the final compost was also determined to assess the fertilizer value. The studies revealed that composting could be applied as a remediation technology after removing the top sediment. The maturity indices that are evaluated from the present study can be used to validate the success of the remediation technology. PMID- 16705826 TI - Trace elements analysis in forage samples from a US Navy bombing range (Vieques, Puerto Rico). AB - Plants are good environmental sensors of the soil conditions in which they are growing. They also respond directly to the state of air. The tops of plants are collectors of air pollutants, and their chemical composition may be a good indicator for contaminated-areas when it is assessed against background values obtained for unpolluted vegetation. Both, aquatic and terrestrial plants are known to bioaccumulate heavy metals and therefore represent a potential source of these contaminants to the human food chain. An evaluation of heavy metals was conducted from vegetation samples collected at the Atlantic Fleet Weapons Training Facilities (AFWTF) in Vieques, Puerto Rico. In order to understand the potential risks associated to heavy metal mobilization through biological systems, it is first necessary to establish background values obtained from reference locations. This information allows a better interpretation of the significance of anthropogenic factors in changing trace elements status in soil and plants. Since Guanica State Forest is located at a similar geoclimatic zone as the AFWTF, samples at this site were used as a standard reference material and as experimental controls. Both sampling and analysis were conducted as previously described in standardized protocols using acid digestion of dry ashes. Then, levels of heavy metals were obtained by air-acetylene flame detection in an atomic absorption spectrophotometer. Our results from the samples taken at the AFWTF indicate mobilization of undesirable trace elements through the marine and terrestrial food web. Since plants naturally remove heavy metals from soils, they could be employed for the restoration of this and similarly contaminated sites. PMID- 16705827 TI - Bacillus subtilis is a potential degrader of pyrene and benzo[a]pyrene. AB - Polycyclic Aromatic Hydrocarbons (PAHs) are a group of compounds that pose many health threats to human and animal life. They occur in nature as a result of incomplete combustion of organic matter, as well as from many anthropogenic sources including cigarette smoke and automobile exhaust. PAHs have been reported to cause liver damage, red blood cell damage and a variety of cancers. Because of this, methods to reduce the amount of PAHs in the environment are continuously being sought. The purpose of this study was to find soil bacteria capable of degrading high molecular weight PAHs, such as pyrene (Pyr) and benzo[a]pyrene (BaP), which contain more than three benzene rings and so persist in the environment. Bacillus subtilis, identified by fatty acid methyl ester (FAME) analysis, was isolated from PAH contaminated soil. Because it grew in the presence of 33 microg/ml each of pyrene, 1-AP and 1-HP, its biodegradation capabilities were assessed. It was found that after a four-day incubation period at 30 degrees C in 20 microg/ml pyrene or benzo[a]pyrene, B. subtilis was able to transform approximately 40% and 50% pyrene and benzo[a]pyrene, respectively. This is the first report implicating B. subtilis in PAH degradation. Whether or not the intermediates resulting from the transformation are more toxic than their parent compounds, and whether B. subtilis is capable of mineralizing pyrene or benzo[a]pyrene to carbon dioxide and water, remains to be evaluated. PMID- 16705828 TI - Degradation of metalaxyl and mefenoxam and effects on the microbiological properties of tropical and temperate soils. AB - The degradation of various formulations of the racemic mixture and the enantiomers (including mefenoxam) of metalaxyl in typical soils from Germany and Cameroon in controlled incubation experiments was studied. The kinetics of the degradation or transformation was determined by means of reversed phase HPLC, while the enantiomeric ratios were measured by HPLC with a chiral Whelk O1 column. The dynamics of the quantitative changes in microbiological properties induced by the addition of these fungicides at their recommended field rates were determined in the soils during a 120-day incubation experiment. The degradation followed first-order kinetics (R > or = 0.96). Higher metalaxyl acid metabolite concentrations were found in German than in Cameroonian soils. The enantiomers of the fungicide had different degradation rates in both soils, with half-lives ranging from 17 to 38 days. All forms of metalaxyl had lower degradation rates in the Cameroonian soil than in the German soil. The degradation of the R-enantiomer was much faster than the S-enantiomer in the German soil and slower than the S enantiomer in the Cameroonian soil, suggesting that different microbial populations, which may be using different enzymes, have different degradation preferences. The type of soil significantly influenced the effect of these fungicides on the soil parameters studied. Incorporation of these fungicides resulted in a change in the ecophysiological status of the soil microbial community as expressed by microbial activities. The activity of phosphatases and fl-glucosidase, the mineralization and availability of N and most plant nutrients in soils were stimulated, whereas the activity of dehydrogenase and the availability of NO3-, were generally adversely affected. The soil NH4+, NO3-, and enzymes activities values in general did not correlate with the degradation of metalaxyl in both soils. However, the degradation of formulated and unformulated metalaxyl was positively correlated to the activity of acid phosphatase in the German soil (R, 0.84 and 0.94 respectively) and in the Cameroonian soil (R, 0.97 and 0.96 respectively). PMID- 16705829 TI - Dietary exposure of fathead minnows to the explosives TNT and RDX and to the pesticide DDT using contaminated invertebrates. AB - Explosive compounds have been released into the environment during manufacturing, handling, and usage procedures. These compounds have been found to persist in the environment and potentially promote detrimental biological effects. The lack of research on bioaccumulation and bioconcentration and especially dietary transfer on aquatic life has resulted in challenges in assessing ecological risks. The objective of this study was to investigate the potential trophic transfer of the explosive compounds 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro 1,3,5-triazine (RDX) using a realistic freshwater prey/predator model and using dichlorodiphenyltrichloroethane (DDT), a highly bioaccumulative compound, to establish relative dietary uptake potential. The oligochaete worm Lumbriculus variegatus was exposed to 14C-labeled TNT, RDX or DDT for 5 hours in water, frozen in meal-size packages and subsequently fed to individual juvenile fathead minnows (Pimephales promelas). Fish were sampled for body residue determination on days 1, 2, 3, 4, 7, and 14 following an 8-hour gut purging period. Extensive metabolism of the parent compound in worms occurred for TNT but not for RDX and DDT. Fish body residue remained relatively unchanged over time for TNT and RDX, but did not approach steady-state concentration for DDT during the exposure period. The bioaccumulation factor (concentration in fish relative to concentration in worms) was 0.018, 0.010, and 0.422 g/g for TNT, RDX and DDT, respectively, confirming the expected relatively low bioaccumulative potential for TNT and RDX through the dietary route. The experimental design was deemed successful in determining the potential for trophic transfer of organic contaminants via a realistic predator/prey exposure scenario. PMID- 16705830 TI - Heavy metal uptake, translocation, and bioaccumulation studies of Triticum aestivum cultivated in contaminated dredged materials. AB - Phytoremediation is a technology that uses vegetation to remediate contaminants from water, soil, and sediments. Unlike traditional remediation techniques such as soil washing or vitrification, phytoremediation offers a technology that is solar-driven, aesthetically pleasing, and cost effective. Recent studies indicate that winter wheat (Triticum aestivum L.) is a potential accumulator for heavy metals such as lead (Pb) and cadmium (Cd) in hydroponic systems. Based on these findings, a laboratory study was conducted with the primary objective of determining the phytoaccumulation capability of this plant species for heavy metals from contaminated dredged materials (DMs) originating from two confined disposal facilities (CDF). The United States Army Corps of Engineers (USACE) manages several hundred million cubic meters of DMs each year, and 5 to 10 % of these DMs require special handling because they are contaminated with hazardous substances that can move from the substrates into food webs causing unacceptable risk outside CDFs. Phytoremediation may offer an alternative to decrease this risk. Chemical analyses by USACE personnel identified 17 metals in various DMs, but in this present study, only zinc (Zn) and Cd were investigated. Pre germinated seeds of the test plants were planted under laboratory conditions in pots containing the various DMs and reference soil. Four weeks after planting, plants were harvested and separated into roots and shoots for biomass production and tissue metal concentrations analyses. Results showed that T. aestivum plants have the capacity to tolerate and grow in multiple-metal contaminated DMs with the potential of accumulating various amounts of Zn and Cd. Root and shoot biomass of T. aestivum were not significantly affected by the DMs on which the plants were grown suggesting that this plant species can grow just as well on DMs contaminated by various metals as in the reference soil. No significant differences in the Zn tissue concentrations were observed, differences in Cd tissue concentrations were noted. A maximum concentration of 26 mg Cd kg(-1) DW was detected in T. aestivum shoots. Although Cd tissue concentrations of T. aestivum plants in this study were below the Cd plant hyperaccumulation criterion of >100 mg kg(-1) Cd found in other studies, this plant species however may still have beneficial uses for phytoremediation studies. T. aestivum plants may serve as an indicator plant for environmental assessment and management, in which the concentration of heavy metals (e.g. Cd) mirrors the concentration in the substrate without dying due to phytotoxicity at low metal concentrations. PMID- 16705831 TI - Morphological and physiological responses of morning glory (Ipomoea lacunosa L.) grown in a lead- and chelate-amended soil. AB - Lead (Pb) is one of the most toxic metals in the environment and may cause drastic morphological and physiological deformities in Ipomoea lacunosa. The goal of this research was to evaluate some morphological and physiological responses of morning glory grown on a Pb- and chelate-amended soil. Soil samples were analyzed, at Mississippi State University Soil Laboratory, for physico-chemical parameters, such as soil texture (73% sand, 23% silt, 4.4% clay), organic matter (6.24 +/- 0.60%), and pH (7.95 +/- 0.03), to establish soil conditions at the beginning of the experiments. Five EDTA (ethylenediaminetetraacetic acid) concentrations (0, 0.1, 0.5, 1, 5mM) and four lead (0, 500, 1000, 2000mg/L) treatments were arranged in factorial in a Randomized Complete Block (RCB) design with five replications. Duncan's multiple comparison range test showed that the mean difference values of stomatal density were significant between 500 and 1000mg/L Pb and between 1000 and 2000mg/L Pb. Two way ANOVA (at 1% level) indicated that interaction between Pb and EDTA had a significant effect on the stomatal density and photosynthetic rates, and at 5% level Pb had a significant effect on chlorophyll concentrations. Lowest concentrations of chlorophyll were recorded at 2000mg/L Pb and 5mM EDTA and exhibited a decreasing trend specifically in the ranges of 1000 and 2000mg/L Pb and 1.0 and 5.0mM EDTA. Duncan's multiple comparison range test confirmed that mean differences between the control treatment vs. 2000mg/L Pb, and 500mg/L vs. 2000mg/L Pb were significantly different atp>0.05. There was a decrease in leaf net photosynthetic rate with increasing concentrations of Pb from 0 to 2000mg/L. In conclusion, I. lacunosa L. plants were grown to maturity in all treatments with no significant and/or apparent morphological disorders, which indicated that this species might be highly tolerant even at 2000mg/L Pb concentrations in the soil. PMID- 16705832 TI - Cytotoxicity of dinitrotoluenes (2,4-dNT, 2,6-DNT ) to MCF-7 and MRC-5 cells. AB - DNTs are considered possibly carcinogenic to humans (Group 2B) because there is inadequate evidence in humans for carcinogenicity though there is sufficient evidence in experimental animals. In this study, MCF-7 (breast) and MRC-5 (lung) cells were exposed to a serial dilution of 2,4 and 2,6 DNTs (control, 1-500 ppm) in 96 well tissue culture plates. After various time intervals (24, 48, 72 and 96 hrs) the plates were washed, and 100microl fluorescein diacetate solution (10 microg/ml in PBS) was added column wise to each well, and incubated at 37 C for 30 - 60 min before reading the fluorescence with a spectrofluorometer at excitation and emission wavelengths of 485 and 538 nm respectively. Spectrofluorometeric readings were converted to percentages of cell survival. Regression analysis was conducted to determine the relationship between cell survival and exposed concentration. Linear equations derived from the regression analysis were used to calculate the LC50 values. Results indicated that 2,6 DNT was more toxic to breast cells; LC50 values were 445 and 292 ppm at 24 and 48 hours respectively compared to 2,4 DNT showing LC50 values of 570 and 407 ppm at 24 and 48 hours, respectively. No significant differences in toxicity existed between the two chemicals with regard to lung cells. Contrary to the above observation, 2,4 DNT was more toxic to breast cells; LC50 values were 407 and 238 ppm at 24 and 48 hours respectively compared to lung cells showing LC50 values of 527 and 402 ppm at 24 and 48 hours respectively. No significant difference existed for 2,6 DNT between the two cell lines. Lungs cells were more resistant to the two chemicals. PMID- 16705833 TI - Changes of multiple metal accumulation (MMA) in New Orleans soil: preliminary evaluation of differences between survey I (1992) and survey II (2000). AB - Soil metal surveys were conducted in Baltimore, MD (1976-1979), Minnesota (1981 1988) and most recently, New Orleans, LA (1989-present). The unique characteristic of New Orleans is that it has two surveys; Survey I was completed in 1992 and Survey II was completed in 2000. This paper seeks to determine if there is a perceptible change in the amount of metals during less than a decade that separated these surveys. The Survey I collection was 4,026 samples stratified by 283 census tracts. All samples were collected in residential neighborhoods at least one block from a busy street. The Survey II collection was 5,467 samples stratified by 286 census tracts (plus City Park). The Survey II collection included busy streets as a category of samples. For comparison, the busy street category of 1,078 samples was excluded from Survey II for a total of 4,388 samples. The extraction methods of the two surveys used the same protocol for strength of acid (1 M HNO3), shaker-time (2 hours), and room temperature (approximately 22 degrees C). However, Survey II differed in amount of sample used in extraction. For Surveys I and II, 4.0g and 0.4g were used respectively. The same ICP-AES was used to measure 8 metals in both surveys. To evaluate the analytical results of the two methods, reference soi 1 samples (n=36) from the Wageningen Evaluating Programs for Analytical Laboratories, International Soil analytical Exchange (WEPAL; ISE) were used. The relationship between the 4.0 and 0.4 g results were linear and the Survey I results were adjusted for sample:acid ratio. Further evaluation was done by creating interpolated Multiple Metal Accumulation (MMA) maps based on the median MMA for each census tract. A new map was created by dividing Survey II MMA by Survey I MMA. The ratio indicates increases of soil metals in the inner city and decreases of soil metals in the outlying areas of Metropolitan New Orleans. Comparing fresh parent alluvium from the Mississippi River with urban soil metal quantities demonstrates that the soils of New Orleans have undergone a massive accumulation of metals. The preliminary results provide ideas about methods needed to further evaluate the changes between these surveys. PMID- 16705834 TI - Impact of animal waste application on runoff water quality in field experimental plots. AB - Animal waste from dairy and poultry operations is an economical and commonly used fertilizer in the state of Louisiana. The application of animal waste to pasture lands not only is a source of fertilizer, but also allows for a convenient method of waste disposal. The disposal of animal wastes on land is a potential nonpoint source of water degradation. Water degradation and human health is a major concern when considering the disposal of large quantities of animal waste. The objective of this research was to determine the effect of animal waste application on biological (fecal coliform, Enterobacter spp. and Escherichia coli) and physical/chemical (temperature, pH, nitrate nitrogen, ammonia nitrogen, phosphate, copper, zinc, and sulfate) characteristics of runoff water in experimental plots. The effects of the application of animal waste have been evaluated by utilizing experimental plots and simulated rainfall events. Samples of runoff water were collected and analyzed for fecal coliforms. Fecal coliforms isolated from these samples were identified to the species level. Chemical analysis was performed following standard test protocols. An analysis of temperature, ammonia nitrogen, nitrate nitrogen, iron, copper, phosphate, potassium, sulfate, zinc and bacterial levels was performed following standard test protocols as presented in Standard Methods for the Examination of Water and Wastewater [1]. In the experimental plots, less time was required in the tilled broiler litter plots for the measured chemicals to decrease below the initial pre treatment levels. A decrease of over 50% was noted between the first and second rainfall events for sulfate levels. This decrease was seen after only four simulated rainfall events in tilled broiler litter plots whereas broiler litter plots required eight simulated rainfall events to show this same type of reduction. A reverse trend was seen in the broiler litter plots and the tilled broiler plots for potassium. Bacteria numbers present after the simulated rainfall events were above 200/100 ml of sample water. It can be concluded that: 1) non-point source pollution has a significant effect on bacterial and nutrients levels in runoff water and in water resources; 2) land application of animal waste for soil fertilization makes a significant contribution to water pollution; 3) the use of tilling can significantly reduce the amount of nutrients available in runoff water. PMID- 16705835 TI - Design and development of low cost, simple, rapid and safe, modified field kits for the visual detection and determination of arsenic in drinking water samples. AB - Arsenic is naturally found in surface and ground waters and the inorganic forms of arsenic are the most toxic forms. The adverse health effects of arsenic may involve the respiratory, gastrointestinal, cardiovascular, nervous, and haematopoietic systems. Arsenic contamination in drinking water is a global problem widely seen in Bangladesh and West Bengal of the Indian sub continent. As there is a great demand for field test kits due to the anticipated reduction of the US EPA arsenic standard from 50ppb to 10ppb a field kit which offers rapid, simple and safe method for precise estimation of arsenic at 10ppb in drinking water samples is developed. Field methods, based on the mercuric-bromide-stain, consist of three different major parts, which are carried out stepwise. The first part of the procedure is to remove serious interference caused by hydrogen sulphide. In commercially available kits either the sulphide is oxidized to sulphate and the excess oxidizing reagent removed prior to the hydride generation step or, the hydrogen sulphide is filtered out by passing the gas stream through a filter impregnated with lead acetate during the hydride generation step. The present method employs cupric chloride in combination with ferric chloride or Fentonis reagent for the removal of hydrogen sulphide, which is rapid, simple and more efficient. Other interferences at this step of the analyses are normally not expected for drinking water analysis. In the second step, the generation of the arsine gas involves the classical way of using zinc metal and hydrochloric acid, which produce the enascenti hydrogen, which is the actual reducing agent. Hydrochloric acid can be replaced by sulfamic acid, which is solid and avoids a major disadvantage of having to handle a corrosive liquid in the field. The arsine gas produces a yellowish spot on the reagent paper. Depending on the arsenic content, either, Yellow n H (HgBr)2 As (10-50ppb), Brown n (HgBr)3 As (50 100ppb) or Black n Hg3 As2 (>100ppb) are formed which can be precisely estimated by visual comparison with standard color chart. The results obtained by field kits agree well with the data obtained through I.C.P.AES methods. The most important characteristic for field measurement is that analytical results can be obtained on the site where the sample is taken with high precision and can be conveniently utilized for monitoring arsenic rapidly in a highly contaminated large geographical area. PMID- 16705837 TI - Comparison assessment of water use and damage between modern and traditional rice irrigation schemes: case of Usangu basin, Tanzania. AB - Water management and competition between users in water scarce river basins is a major challenge facing the human race. The inter dependence of users in such basins, necessitates a clear understanding of each user in relation to the location, the water demand, and the duration of water need. The understanding of these factors, together, is very important for the management of water resources in such basins without which, it is argued that, water is overused and wasted. As an example of this, the large modern and improved rice irrigation systems in Tanzania are believed to use water more efficient than the traditional irrigation systems. Yet, well-founded scientific analyses are a necessary part to quantify such beliefs as they can inform us whether the natural resource, in such systems, is properly utilized and managed or not. Likewise, such studies can allow us to quantify how much water is over used and thus the natural resource is unnecessarily degraded. This paper explores a study conducted in the Usangu basin, Tanzania, to investigate the gross and net needs for modern and traditional rice irrigation schemes, and the implications with regards to water resource management and damage. Problems relating to modernization of traditional smallholder irrigation systems and upstream--downstream water users are further discussed. The paper concludes from the study that modern irrigation schemes are inefficient compared to traditional irrigation schemes. Also modernization of traditional schemes in the study area have resulted into over abstraction and reduced productivity of water. Looking to the future, this study tells us that improvement or modernization of irrigation infrastructure should be balanced between negative impacts to available water resources albeit it's significant economic contribution to the community. PMID- 16705836 TI - Assessing interactions of multiple agrichemicals by using bacterial assemblages in a wetland mesocosm system. AB - Agrichemicals may enter wetlands located adjacent to or downstream from agricultural fields. We investigated the individual and interactive effects of three agrichemicals [atrazine, chlorpyrifos, and monosodium acid methanearsonate (MSMA)] and methyl mercury on abundance and heterotrophic potential of wetland heterotrophic bacteria assemblages. We used a factorial experimental design, in which chemicals were introduced in all possible combinations to 66 500-liter mesocosms at the Biological Field Station of the University of Mississippi. Methyl mercury was added to bring the total mercury (HG) concentration to 0.4 mg/Kg wet weight at the sediment surface. Atrazine, chlorpyrifos, and MSMA were added at concentrations of 192, 51, and 219 microg/L, respectively. Over 32 days of exposure, microbial heterotrophic activity was sensitive to only the interactive effect of HG*ATR*CPF in the sediments and only CPF in the water. Total bacterial numbers did not exhibit any significant treatment effects. Therefore, the effects of agrichemicals were reflected on cell-specific bacterial heterotrophic activity rather than bacterial population size. PMID- 16705838 TI - Use of multi-objective air pollution monitoring sites and online air pollution monitoring system for total health risk assessment in Hyderabad, India. AB - A consensus has been emerging among public health experts in developing countries that air pollution, even at current ambient levels, aggravates respiratory and cardiovascular diseases and leads to premature mortality. Recent studies have also presented well-founded theories concerning the biological mechanisms involved and the groups of people that are probably more susceptible to health effects caused or exacerbated by inhalation of ambient particulate matter (PM.). On the basis of prognostic studies carried out in Center for Environment, JNT University, Hyderabad "it has been estimated that in Hyderabad some 1,700 to 3,000 people per year die prematurely as a result of inhaling PM". These figures reflect only the effects of acute exposure to air pollution. If the long-term effects of chronic exposure are taken into account, 10,000-15,000 people a year could die prematurely in Hyderabad. This estimate of the chronic effects is based on other studies, which are not completely comparable with the Hyderabad situation. While the study designs and analyses in these other studies may indeed be different or irrelevant to Hyderabad, the fact they were carried out in other countries is irrelevant. Taking into account these considerations, a model for total health risk assessment for the city of Hyderabad, and its state of Andhra Pradesh in India has been developed using a multi-objective air pollution monitoring network and online and real time air pollution monitoring stations. For the model studies a number of potential monitoring sites were screened for general and site-specific criteria in a geographic information system (GIS) environment that may, on a local basis, affect the representativeness of the data collected. Local features that may affect either the chemical or meteorological parameters are evaluated to assure a minimum of interference. Finally, for monitoring air pollution, an online and real-time monitoring system was designed using advanced electrochemical sensor systems (sulphur dioxide, oxides of nitrogen, carbon monoxide, hydrocarbons, ozone, mercaptans and hydrogen sulphide) and a particulate matter analyzer (total suspended particulate matter TSPM), PM2.5 and PM10). The sensor and data acquisition systems are programmed to monitor pollution levels at 1/2 hour durations during peak hours and at 1-hour intervals at other times. Presently, extensive statistical and numerical simulations are being carried out at our center to correlate the individuals living in the monitored areas with respiratory infections with air pollution. PMID- 16705840 TI - The seasonality of fecal coliform bacteria pollution and its influence on closures of shellfish harvesting areas in Mississippi Sound. AB - Runoff from agricultural lands and farm animal feedlots is one of the major sources of fecal coliforms in surface waters, and fecal coliform (FC) bacteria concentrations tend to vary with season because of seasonal variations in climatic factors. However, El Nino--Southern Oscillation (ENSO) events may affect the extent and patterns of seasonality in FC levels in coastal waters. Water quality monitoring data for shellfish management collected during El Nino (1990, 1992, 1997), and La Nina (1999, 2000) years were analyzed to evaluate the extent to which these events influenced Pearl River stage, and bacterial levels, water temperature, and salinity in the western part of Mississippi Sound. Models to predict FC levels in relation to various environmental factors were also developed. In 1990, 1992 and 1997, FC geometric mean counts peaked in late winter (January/February) reaching 120 MPN (February 1990), 165 MPN (January 1992), and 86 MPN (January 1997), and then decreased considerably during spring and summer (1.2 - 19 MPN). Thereafter, FC abundance increased slightly in fall and early winter (1.9 - 24 MPN). Fecal coliform abundance during the 2000 La Nina year was much lower (1.0 -10.3 MPN) than in 1992 (1.2 - 165 MPN), and showed no seasonal pattern from January to August, perhaps due to the relative scarcity of rainfall in 2000. In 1995 (ENSO neutral year), peak geometric mean FC count (46 MPN) was lower than during El Nino years and occurred in early spring (March). The seasonal and between year variations in FC levels determined the number of days during which the conditionally approved shellfish growing area was opened for harvesting shellfish. For example, from January to April 1997, the area was not opened for shellfish harvesting, whereas in 2000, the number of days during which the area was opened ranged from 6 - 27 (January to April) to 24 - 26 (October to December). ENSO events thus influenced the extent and timing of the peak levels of fecal coliforms in Mississippi Sound. Models consisting of one or more of the variables: Pearl River stage, water temperature, and salinity were developed to predict FC concentrations in the Sound. The model parameter(s) explained 56 to 91% of the variations in FC counts. Management of shellfish in Mississippi Sound can be improved by utilizing information on the forecasted three to seven years occurrence of ENSO events. In addition, since Pearl River stage was the most important variable predicting FC concentration in the Sound, a study of the levels and sources of FC bacteria in the river, especially the middle and lower sections, is needed for developing a management plan for reducing FC bacteria pollution in the Sound. PMID- 16705839 TI - Cytotoxicity and expression of c-fos, HSP70, and GADD45/153 proteins in human liver carcinoma (HepG2) cells exposed to dinitrotoluenes. AB - Dinitrotoluenes (DNTs) are byproducts of the explosive trinitrotoluene (TNT), and exist as a mixture of 2 to 6 isomers, with 2,4-DNT and 2,6-DNT being the most significant. The main route of human exposure at ammunition facilities is inhalation. The primary targets of DNTs toxicity are the hematopoietic system, cardiovascular system, nervous system and reproductive system. In factory workers, exposure to DNTs has been linked to many adverse health effects, including: cyanosis, vertigo, headache, metallic taste, dyspnea, weakness and lassitude, loss of appetite, nausea, and vomiting. Other symptoms including pain or parasthesia in extremities, abdominal discomfort, tremors, paralysis, chest pain, and unconsciousness have been documented. An association between DNTs exposure and increased risk of hepatocellular carcinomas and subcutaneous tumors in rats, as well as renal tumors in mice, has been established. This research was therefore designed targeting the liver to assess the cellular and molecular responses of human liver carcinoma cells following exposure to 2,4-DNT and 2,6 DNT. Cytotoxicity was evaluated using the MTT assay. Upon 48 hrs of exposure, LC50 values of 245 +/- 14.724 microg/mL, and 300 +/- 5.92 microg/mL were recorded for 2,6-DNT and 2,4-DNT respectively, indicating that both DNTs are moderately toxic, and 2,6-DNT is slightly more toxic to HepG2 cells than 2,4-DNT. A dose response relationship was recorded with respect to the cytotoxicity of both DNTs. Western blot analysis resulted in a significant expression (p<0.05) of the 70-kDa heat shock protein in 2,6-DNT-treated cells compared to the control cells and at the 200 microg/mL dose for 2,4-DNT. A statistically significant expression in c fos was also observed at the 200 and 250 microg/mL treatment level for 2,4- and 2,6-DNT, respectively. However, no statistically significant expression of this protooncogene-related protein was observed at the doses of 0, 100, or 300 microg/mL or within the dose range of 0-200 microg/mL for 2,6-DNT. The 45-kDa growth arrest and damage protein was significantly expressed at the dose range of 200 - 250 microg/mL for 2,6-DNT and at the dose range of 200 - 400 microg/mL for 2,4-DNT. Expression of 153-kDa growth arrest and DNA damage protein was significant at the 100, 200, and 250 microg/mL doses for 2,6-DNT and at the 200 microg/mL dose for 2,4-DNT. Overall, these results indicate the potential of DNTs to induce cytotoxic, proteotoxic (HSP70), and genotoxic (GADD45/153) effects, as well as oxidative stress and pro-inflammatory reactions (c-fos). PMID- 16705841 TI - A multicentral randomized control study on clinical acupuncture treatment of Bell's palsy. AB - OBJECTIVE: To confirm the clinical therapeutic effects of acupuncture and moxibustion on Bell's palsy. METHODS: 480 cases from 4 hospitals were enrolled for this study, among whom 439 cases completed the whole course of the study. The patients were randomly divided into the following 3 groups, a control group(treated with prednisone, vitamin B1, vitamin B12 and dibazol), an acu moxibustion group (treated with filiform needle plus moxibustion), and a basic treatment plus acu-moxibustion group (treated with oral medicine like those in the control group plus acupuncture, and with moxibustion like in the acu moxibustion group). The whole treatment course lasted 4 weeks. The therapeutic effects were evaluated according to the symptoms and signs, House-Brackmann grading scale and facial disability indexes(FDI). RESULTS: All the 4 centers (hospitals) completed this study well, with no statistically significant difference found among the 4 centers in therapeutic effects. The patients with different conditions were well distributed in the 3 groups, thus the basic general data were comparable (P > 0.05). The therapeutic effects of the two treatment groups were better than the control group (respectively P < 0.05 and P < 0.01), and it was the best in the acu-moxibustion group (P < 0.01). CONCLUSION: Acupuncture and moxibustion may exert definite therapeutic effects on Bell's palsy, better than that of the basic treatment group or the basic treatment plus acu-moxibustion group. PMID- 16705842 TI - Effects of cupping therapy on the pulmonary functions in asthmatic children. PMID- 16705843 TI - Clinical study on the wrist-ankle acupuncture treatment for 30 cases of diabetic peripheral neuritis. AB - OBJECTIVE: To study the mechanisms of wrist-ankle acupuncture for prevention and treatment of diabetic peripheral neuritis. METHODS: Ninety cases of diabetic peripheral neuritis were randomly divided into 3 groups, and treated respectively with wrist-ankle acupuncture, body-acupuncture, and the western routine medical treatment, with 30 cases in each of the groups; and therapeutic effects and laboratory results compared. RESULTS: It is proved that the therapeutic effects of the wrist-ankle acupuncture group and body acupuncture group were significantly superior to those of the control group, with no significant differences between the former two groups. CONCLUSION: Wrist-ankle acupuncture has the actions of improving the metabolisms of blood sugar and blood-lipid, lowering down blood viscosity, and restoring the functions of peripheral nerve cells, thus giving definite therapeutic effects for diabetic peripheral neuritis. PMID- 16705844 TI - Acupuncture treatment of acute cerebral infarction by the 'Three-Step Needling' method. AB - OBJECTIVE: To observe the therapeutic effects of acupuncture treatment by the 'Three-Step Needling' Method for acute cerebral infarction. METHOD: While receiving the routine medical treatment, 68 cases were treated by the acupuncture method of 'Three-Step Needling'. RESULTS: 27 of the 68 cases were basically cured, 23 markedly improved, 15 improved, and 3 failed, the total effective rate being 95.59%. CONCLUSION: The acupuncture method of 'Three-Step Needling' can give quite good therapeutic effects for acute cerebral infarction. PMID- 16705845 TI - Jinger moxibustion for treatment of cervical vertigo --a report of 40 cases. AB - In order to observe the therapeutic effects of jinger moxibustion for cervical vertigo, 78 cases were randomly divided into 2 groups, of whom 40 were treated with jinger moxibustion and 38 treated with acupuncture. The results showed a cure rate of 72.5% with a total effective rate of 97.5% in the jinger moxibustion group, while 44.7% and 73.7% respectively in the acupuncture group. The comparisons between the two groups in the cure rate and the total effective rate showed significant differences (P < 0.05). THE CONCLUSION: Jinger moxibustion has a good therapeutic effect for treating cervical vertigo, superior to that of the acupuncture treatment. PMID- 16705846 TI - The TCM-combined treatment for aphasia due to cerebrovascular disorders. AB - OBJECTIVE: To evaluate the therapeutic effects of scalp acupuncture (with the cluster needling, a long needle-retention and an intermittent manipulation) combined with the Schuell's stimulation and psychological care for treatment of aphasia due to cerebrovascular disorders. METHOD: 36 eligible cases of aphasia were randomly assigned into a treatment group and a control group. The scoring system for assessment of aphasia in speaking Chinese set by CMA Neurological Branch and that of BADE were adopted for grading the severity/degree of aphasia before and after the treatment. RESULTS: The total effective rate in the treatment group was 84.21%, and that in the control group was 70.59%, with a very statistically significant difference (P < 0.01). CONCLUSION: The combined scheme produced a better therapeutic effect. PMID- 16705847 TI - Acupuncture plus point-injection for 32 cases of obstinate urticaria. AB - In order to observe the therapeutic effects of acupuncture plus point-injection for obstinate urticaria, 64 cases of obstinate urticaria were randomly divided into the following two groups. 32 cases in the treatment group were treated with acupuncture at the points of Quchi (LI11), Xuehai (SP10), Zusanli (ST 36), Sanyinjiao (SP6) and Fengchi (GB20) plus point-injection at the points of Zusanli (ST36) and Quchi (LI11). 32 cases in the control group were treated with antihistamines (such as Acrivastine, Cinnarizine or Ranitidine). The results showed that the therapeutic effect in the treatment group was obviously better than that in the control group (P < 0.05), with a much lower relapse rate in the former than that in the latter (P < 0.01). PMID- 16705848 TI - Forty cases of simple obesity treated by acupuncture. PMID- 16705849 TI - Treatment of ankylosing spondylitis with medicated moxibustion plus salicylazosulfapyridine and methotrexate--a report of 30 cases. AB - OBJECTIVE: To evaluate the therapeutic effect of medicated moxibustion plus administration of salicylazosulfapyridine (SASP) and methotrexate (MTX) for treatment of active ankylosing spondylitis (AS). METHOD: Ninety cases of active AS were randomly assigned to Group A, B and C (N = 30 in each group), and treated respectively with SASP and MTX, acupuncture plus SASP and MTX, and composite sulfur (tablet) moxibustion plus SASP and MTX for 3 successive courses (2 months each course with an interval of 5 days). RESULTS: Improvement in sacroiliitis index, Schober test, occipital wall test, finger-ground distance, as well as the erythrocyte sedimentation rate (ESR) and the content of C-reactive protein (CRP) in Group B and C was far superior to that of Group A (P < 0.01). CONCLUSION: Combined use of western medicine with acupuncture or with medicated moxibustion produces a better therapeutic effect than western medicine given alone. PMID- 16705850 TI - The therapeutic effects of acupuncture in 30 cases of postpartum hypogalactia. PMID- 16705851 TI - Combined use of acupuncture and cupping for emergency treatments. PMID- 16705852 TI - Fufang cangzhu tang for treatment of senile obesity or overweight complicated with impaired glucose tolerance --a clinical observation in 32 cases. AB - OBJECTIVE: To observe the therapeutic effect of Fufang Cangzhu Tang (Composite Atractylodes Decoction) on senile obesity or overweight with impaired glucose tolerance (IGT). METHODS: 32 cases of senile obesity or overweight with IGT were treated with Composite Atractylodes Decoction, with another 30 cases treated with dimethyldiguanide as the controls. Changes of body weight, waist circumference, hip circumference, waist hip circumference ratio (WHR), glucose tolerance, fast serum insulin and blood lipid before and after treatment were compared. RESULTS: After treatment, the body weight, waist circumference, hip circumference and WHR, glucose tolerance, fast serum insulin and blood lipid in the Composite Atractylodes Decoction treatment group decreased significantly (P < 0.05 or P < 0.01), with no significant difference as compared with the control group (P > 0.05). CONCLUSION: Composite Atractylodes Decoction can obviously decrease the body weight, waist circumference, hip circumference, WHR, glucose tolerance, fast serum insulin and blood lipid in the senile patients with obesity or overweight with impaired glucose tolerance. PMID- 16705853 TI - TCM treatment of male immune infertility--a report of 100 cases. AB - OBJECTIVE: To observe the therapeutic effect of Yikang Tang (Yikang Decoction) for male immune infertility. METHODS: 100 cases of male immune infertility in the treatment group were treated with Yikang Decoction, while 100 cases treated with prednisone as the controls. Physical exam, routine semen and prostate exams, and exams for presence of anti-sperm antibody (AsAb) and mycoplasma in the serum or seminal plasma were carried out. RESULTS: 1) The serum and seminal plasma AsAb levels decreased significantly (P < 0.01) in both the groups after treatment, with a more remarkable effect in the treatment group. 2) The sperm density and percentage of motile spermatozoa increased significantly in the two groups, but more significantly in the treatment group after treatment. The pregnancy rate of their wives was higher in the treatment group than that in the control group (P < 0.01). 3) The sperm agglutination rate in the two groups decreased, but more significantly in the treatment group after treatment. 4) The improvement rate of the symptoms and the stability of the therapeutic effect were more dramatic in the treatment group than that in the control group (P < 0.01) after termination of drug administration. CONCLUSION: The Yikang Decoction has a more stable effect for male immune infertility than prednisone. PMID- 16705854 TI - Shugan jieyu yin for treatment of senile depression--a clinical report of 84 cases. PMID- 16705855 TI - Dr. Zhang Ren's experience in acupuncture treatment of obstinate eye diseases. PMID- 16705856 TI - Dr. Yang Nizhi's experience in treating benign arteriolar nephrosclerosis. PMID- 16705857 TI - Dr. Lu Zhizheng's experience in treating sicca syndrome. PMID- 16705858 TI - Effects of taohong siwu decoction II in the chick chorioallantoic membrane (CAM) assay and on B16 melanoma in mice and endothelial cells ECV304 proliferation. AB - OBJECTIVE: To investigate the anti-angiogenesis action of Taohong Siwu Decoction II (THSWD II). METHODS: The chick chorioallantoic membrane (CAM) assay was adopted to study the anti-angiogenesis action of THSWD II; the MTT test was used to investigate its effect on proliferation of the human umbilical vein endothelial cells ECV304; and the immunohistochemical method was used to observe the effect of THSWD II on the expression of kinase insert domain containing receptor/fetal liver kinase 1 (KDR/Flk-1) and the microvessel density (MVD) of B16 melanoma in mice. RESULTS: After treatment with THSWD II, the blood vessel index of CAM and the absorbency of ECV304 in the THSWD II 1 mg/ml group and the 2 mg/ml group decreased significantly (P < 0.01); the weight, the expression of KDR/Flk-1 and the MVD of B16 melanoma in mice reduced significantly in the THSWD II 5 g/kg group, the 10 g/kg group and the TSHSWD 10g/kg plus cyclophosphamide group (P < 0.01). CONCLUSION: THSWD II has the actions of anti-angiogenesis, and inhibiting the proliferation of ECV304 cells and the growth of B16 melanoma. The clinical anti-tumour mechanism is considered to berelated possibly to its anti angiogenesis action by inhibiting the expression of KDR/FIK-1. PMID- 16705859 TI - Effects of extract of dilong (pheretima) on the scalded skin in rats. AB - In order to investigate the effects of extract of Dilong (Pheretima) on the skin viability and lipid peroxidation after deep degree II scalds, the extract of Dilong (Pheretima) at two concentrations (5%, 10%) were respectively applied to the surface of wound. The oxygen consumption, the succinate dehydrogenase (SDH) activity, the Schiff's base content, and the healing time of wound surface were determined in Wistar rats at the 8th, 24th, 48th and 72th hour after scalds, with SD-Ag used as controls. The results indicated that the oxygen consumption and SDH activity decreased, and the schiff's base content increased after the scalds. After the application of the extract of Dilong, the oxygen consumption and SDH activity increased, and the schiff's base content decreased significantly as compared with the control group (P < 0.05 or P < 0.01), and the mean healing time of the wound surface was five days in advance in the two treatment groups as compared with the control group. It is suggested that the scalds are related with lipid peroxdation to a certain extent, and the extract of Dilong has actions of attenuating lipid peroxidation, and promoting healing of the wound. PMID- 16705860 TI - Effects of Chinese herbal medicine based on hachimi-jio-gan on osteopenia in rats. AB - The effects of Chinese herbal medicines including Hachimi-jio-gan (HJG) and/or Hochu-ekki-to (HEW) on osteopenia in rats were investigated. The Chinese herbal medicines were administrated for 8 weeks (7 times/week) starting from 1 week after ovariectomy. HJG and Prescription-2 (Prsc-2, the prescription based on HJG) showed protective effect on bone loss of the vertebrae after ovariectomy. However, Prescription-1 (the prescription based on HJG and HET) had no effect. Then, we made osteopenia model in rats by prednisolone and low calcium diet. Prsc 2 and HJG were administrated for 20 weeks with prednisolone. These Chinese remedies showed protective effects for osteopenia, with better indices on bone loss of the limbs than HJG alone in the osteopenia rats. It can be concluded that Prsc-2 is more effective than HJG for bone loss induced by various factors, and the additives in Prsc-2 may enhance the therapeutic effect. PMID- 16705861 TI - Acupuncture treatment of shoulder pain. PMID- 16705862 TI - Brief introduction to digital acupoint pressure therapy. PMID- 16705863 TI - Forced dissociation of selectin-ligand complexes using steered molecular dynamics simulation. AB - Selectin-ligand interactions are crucial to such biological processes as inflammatory cascade or tumor metastasis. How transient formation and dissociation of selectin-ligand bonds in blood flow are coupled to molecular conformation at atomic level, however, has not been well understood. In this study, steered molecular dynamics (SMD) simulations were used to elucidate the intramolecular and intermolecular conformational evolutions involved in forced dissociation of three selectin-ligand systems: the construct consisting of P selectin lectin (Lec) and epidermal growth factor (EGF)-like domains (P-LE) interacting with synthesized sulfoglycopeptide or SGP-3, P-LE with sialyl Lewis X (sLe(X)), and E-LE with sLe(X). SMD simulations were based on newly built-up force field parameters including carbohydrate units and sulfated tyrosine(s) using an analogy approach. The simulations demonstrated that the complex dissociation was coupled to the molecular extension. While the intramolecular unraveling in P-LE-SGP-3 system mainly resulted from the destroy of the two anti parallel beta sheets of EGF domain and the breakage of hydrogen-bond cluster at the Lec-EGF interface, the intermolecular dissociation was mainly determined by separation of fucose (FUC) from Ca2+ ion in all three systems. Conformational changes during forced dissociations depended on pulling velocities and forces, as well as on how the force was applied. This work provides an insight into better understanding of conformational changes and adhesive functionality of selectin ligand interactions under external forces. PMID- 16705864 TI - Misfolding dynamics of human prion protein. AB - We report the results of longest to date simulation on misfolding of monomeric human prion protein (HuPrP). By comparing our simulation of a partially unfolded protein to the simulation of the native protein, we observe that the native protein as well as native regions in the partially unfolded protein remain in the native state, and the unfolded regions fold back with increased extended (sheet and PP-II) conformations. The misfolded regions show increased basin hopping from non-helical basins while the amino acids locked in the helical conformation tend to stay locked in that conformation. Our results also validate the hypothesis that denaturation of helices and formation of a partially unfolded intermediate is required for misfolding as the native protein stayed in native conformation for the entire simulation. Finally, we also observe that there is no correlation between misfolding and the chemical identity of amino acids, as both hydrophobic and hydrophilic amino acids showed equal probability of sampling extensively from non-native conformations. PMID- 16705865 TI - The effect of matrix tension-compression nonlinearity and fixed negative charges on chondrocyte responses in cartilage. AB - Thorough analyses of the mechano-electrochemical interaction between articular cartilage matrix and the chondrocytes are crucial to understanding of the signal transduction mechanisms that modulate the cell metabolic activities and biosynthesis. Attempts have been made to model the chondrocytes embedded in the collagen-proteoglycan extracellular matrix to determine the distribution of local stress-strain field, fluid pressure and the time-dependent deformation of the cell. To date, these models still have not taken into account a remarkable characteristic of the cartilage extracellular matrix given rise from organization of the collagen fiber architecture, now known as the tension-compression nonlinearity (TCN) of the tissue, as well as the effect of negative charges attached to the proteoglycan molecules, and the cell cytoskeleton that interacts with mobile ions in the interstitial fluid to create osmotic and electro-kinetic events in and around the cells. In this study, we proposed a triphasic, multi scale, finite element model incorporating the Conewise Linear Elasticity that can describe the various known coupled mechanical, electrical and chemical events, while at the same time representing the TCN of the extracellular matrix. The model was employed to perform a detailed analysis of the chondrocytes' deformational and volume responses, and to quantitatively describe the mechano electrochemical environment of these cells. Such a model describes contributions of the known detailed micro-structural and composition of articular cartilage. Expectedly, results from model simulations showed substantial effects of the matrix TCN on the cell deformational and volume change response. A low compressive Poisson's ratio of the cartilage matrix exhibiting TCN resulted in dramatic recoiling behavior of the tissue under unconfined compression and induced significant volume change in the cell. The fixed charge density of the chondrocyte and the pericellular matrix were also found to play an important role in both the time-dependent and equilibrium deformation of the cell. The pericellular matrix tended to create a uniform osmolarity around the cell and overall amplified the cell volume change. It is concluded that the proposed model can be a useful tool that allows detailed analysis of the mechano-electrochemical interactions between the chondrocytes and its surrounding extracellular matrix, which leads to more quantitative insights in the cell mechano-transduction. PMID- 16705866 TI - Intracellular stress transmission through actin stress fiber network in adherent vascular cells. AB - Intracellular stress transmission through subcellular structural components has been proposed to affect activation of localized mechano-sensing sites such as focal adhesions in adherent cells. Previous studies reported that physiological extracellular forces produced heterogeneous spatial distributions of cytoplasmic strain. However, mechanical signaling pathway involved in intracellular force transmission through basal actin stress fibers (SFs), a mechano-responsive cytoskeletal structure, remains elusive. In the present study, we investigated force balance within the basal SFs of cultured smooth muscle cells and endothelial cells by (i) removing the cell membrane and cytoplasmic constituents except for materials physically attaching to the substrate (i.e., SF-focal adhesion complexities) or (ii) dislodging either mechanically or chemically the cell processes of the cells expressing fluorescent proteins-labeled actin and focal adhesions in order, to examine stress-release-induced deformation of the basal SFs. The result showed that a removal of mechanical restrictions for SFs resulted in a decrease in the length of the remaining SFs, which means SFs bear tension. In addition, a release of the preexisting tension in a single SF was transmitted to another SF physically linked to the former, but not transmitted to the other ones physically independent of the former, suggesting that the prestress is balanced in tensed SF networks. These results support a hypothesis regarding cell structural architecture that physiological extracellular forces can produce in the basal SF network a directional intracellular stress or strain distribution. Therefore, consideration of the coexistence of the directional stretching strain along the axial direction of SFs and the heterogeneous strain in the other cytoplasmic region will be essential for understanding intracellular stress transmission in the adherent cells. PMID- 16705868 TI - Hormonal therapy in menopausal women. PMID- 16705867 TI - Systolic modeling of the left ventricle as a mechatronic system: determination of myocardial fiber's sarcomere contractile characteristics and new performance indices. AB - BACKGROUND: In this paper, the left ventricle (LV) is modeled as a cylinder with myocardial fibers located helically within its wall. A fiber is modeled into myocardial structural units (MSUs); the core entity of each MSU is the sarcomeric contractile element. The relationship between the sarcomere unit's contractile force and shortening velocity is expressed in terms of the LV model's wall stress and deformation, and hence in terms of the monitored LV pressure and volume. Then, the LV systolic performance is investigated in terms of a mechatronic (excitation-contraction) model of the sarcomere unit located within the LV cylindrical model wall. METHODS: The governing equation of dynamics of the LV myocardial structural unit (MSU) is developed, involving the parameters of the series-elastic element (SE), the viscous element (VE) and the contractile element (CE). We then relate the MSU's force and displacement variables (in terms of SE, VE and CE parameters) to the LV pressure and volume, using the patient's catheterization-ventriculogram data. We thereby evaluate the MSU elements' parameters. RESULTS: We then determine the sarcomere (CE) 'force vs. shortening velocity' characteristics as well as the power generated by the sarcomere (or CE) element. These are deemed to be important LV functional indices. When our computed sarcomeric peak-power is compared against the traditional LV contractility indices (by linear regression), a high degree of correlation is obtained. CONCLUSIONS: We have provided herein, a LV systolic-phase (cylindrical geometry) model whose wall contains the myocardial fibers having sarcomere units. We have expressed the LV myocardial sarcomere's CE (force vs. shortening velocity) characteristics in terms of the LV pressure-volume data. These CE properties express the intrinsic performance capacity of the LV. Hence, indices containing these properties are deemed to reflect LV performance. In this regard, our new LV contractility index correlates very well with the traditional LV contractility index dP/dt(max). PMID- 16705869 TI - President's pen. PMID- 16705870 TI - Guidelines for the registered nurse in accepting or rejecting a work assignment. PMID- 16705871 TI - What is the Vision Paper: the future regulation of advanced practice nursing? PMID- 16705872 TI - Improving the world's health--the role of National Public Health Institutes. PMID- 16705873 TI - The evaluation of short and long-term administration of drugs improving life expectancy in patients with myocardial infarction in a Hungarian county hospital. AB - OBJECTIVE: The patients after myocardial infarction could hope for a significant improvement in their life expectancy when complying with the principles of the secondary prevention. Today there is no doubt that the administration of ACE inhibitors, beta-blockers, aspirin and statins decrease mortality in these patients. Although the clinicians are aware of the guidelines of Evidence Based Medicine, international and Hungarian surveys show that in their everyday application there is still much to improve. PATIENTS AND METHODS: The authors studied the therapy of 200 consecutive patients (115 men and 85 women) who suffered from acute myocardial infarction (either STAMI or NSTAMI) in 1999-2000 at the discharge from their internal medicine department with cardiological profile and 6 months later during outpatient check-up. Having these therapeutic data a special emphasise was given to the same group of drugs and their administration was investigated in patients suffering from acute coronary syndrome with elevated troponin-T levels in the year 2002. RESULTS: In the study of years 1999-2000 at the discharge 175 of 200 patients received ACE-inhibitor (87%), 121 (60%) beta-blocker, 180 (90%) antiplatelet and 102 (51%) statin therapy. At the time of the control performed 6 months later 85% of the patients were on ACE-inhibitor, 88% on beta-blocker, 77% on aspirin and 47% on statin therapy. In this high risk population during the 6 months control the LDL cholesterol goal of 2.5 mmol/l was attained in the 17% of patients. In the survey performed in 2002 the use of beta-receptor blocker increased to 85%, antiplatelet drug and statin administration to 95% and 57%, respectively. CONCLUSIONS: Although the administration of drugs improving life expectancy in the authors' department is comparable with the published Hungarian and international data, effort to the widespread application of the ever growing principles of the Evidence Based Medicine and continuous self-control are essential. PMID- 16705874 TI - Pregnancy complications and delivery outcomes of pregnant women with common cold. AB - OBJECTIVE: To study the association between common cold during pregnancy and pregnancy complications and delivery outcomes: gestational age/birth weight, in addition preterm birth and low birthweight. METHOD: In the population-based large data set of the Hungarian Case-Control Surveillance System of Congenital Abnormalities (HCCSCA), 1980-1996, controls without congenital abnormalities were analysed. RESULTS: Of 38,151 newborn infants, 5,475 (14.4%) had mothers with common cold. The prevalence of threatened preterm delivery, placental disorders and severe nausea and vomiting was lower while the occurrence of anemia was higher in pregnant mothers with common cold than in mothers without common cold. Mothers with common cold in pregnancy had 0.1 week shorter gestational age, thus the proportion of preterm births (9.8% vs 9.1%) was somewhat larger. However, mean birth weight was somewhat larger (3,305 vs 3,271 g) and the proportion of low birthweight newborns (4.2% vs 5.9%) was smaller. CONCLUSION: Common cold during pregnancy does not increase the occurrence of pregnancy complications except anemia, while delivery outcomes showed minor but opposite (higher rate of preterm birth and lower rate of low birthweight) changes. PMID- 16705875 TI - Dyslipidemia in industrial workers in hot environments. AB - The aim of the study was to follow the rate and manifestation of dyslipidemia in industrial workers exposed to heat. One hundred and two male industrial workers exposed to heat and a control group of 102 male workers were studied. The microclimate components were followed and Wet Bulb Globe Temperature (WBGT) was calculated. The mean WGBT was 35.4 degrees C (28.4-41.7 degrees C) for the studied heat exposed work places. The lipid indices: total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL C) and triacylglycerols were assayed with enzymatic tests. TC/HDL-C ratio was calculated, too. Arterial pressure, anthropometric variables, smoking, alcohol use were followed and no significant differences between the studied groups were found. Significantly higher TC, LDL-C and TC/HDL-C were found with the heat exposed industrial workers. Odds ratio indicated higher risk in heat exposed industrial workers of becoming dyslipidemic [for TC OR = 1.481 (1.097-2.002) and for LDL-C OR = 1.539 (1,123-2.111)]. Regular screening of lipid profile in heat exposed workers is recommended. PMID- 16705876 TI - Dietary intake of sulphites by children in the Slovak Republic. AB - The mean dietary intake of sulphites by Slovak children aged 7 to 10 years was estimated. Calculations were based on a food model of a specific boarding-school in January, May, July and October while respecting the scientifically recommended dietary allowances. It was assumed that sulphites were used in the widest possible range of foods and at maximum permitted levels. Due to this fact and since sulphites are partially lost during food processing and storage, the results are over-estimated. However the mean daily exposure of consumers with the lowest (21 kg) and the mean (26.5 kg) body weight to sulphites ranged from 44 to 55% and from 36 to 43% of the acceptable daily intake value (ADI) respectively. Due to availability of various foods the dietary models were not affected seasonally and thus no differences among intakes of sulphites in individual months were obvious. The results indicated that the above mentioned group of children is sufficiently protected from the hazard of sulphites in food. PMID- 16705877 TI - Serological profile and virological evaluation of hepatitis B and hepatitis C virus infection among HIV infected patients in Greece. AB - In this study we evaluate the prevalence of HBV and HCV infections and the HBV and/or HCV viral load as well as HCV genotype among 737 HIV-infected patients. 89/737 (12.1%) were HBsAg(+) and the majority of them (60.7%) were HBeAg(+), in contrast to general Greek population; anti-HBc seropositivity was detected in 48.1% of the study population. Serum HBV-DNA levels were 5.75 +/- 1.66 (-log 10 copies/ml) and HBeAg(+) coinfected patients had significantly higher levels than HBeAg(-) ones (7.40 +/- 0.64 vs 4.59 +/- 1.01, respectively, p < 0.001). 8.2% of HIV-infected patients were anti-HCV(+) and the majority of them (85.7%) had HCV RNA levels more than 700.000 IU/I. The most common HCV-genotype was genotype-1 (12/28, 42.9%), representing a difficult-to-treat special population. PMID- 16705878 TI - Privatisation of hospitals in Poland--barriers and opportunities. AB - The paper discusses the question of privatisation of the in-patient care sector in Poland. The references to the general reforming processes are made, as well as the legal context of ownership changes is presented. First part of the article describes the forms of privatisation applied in Poland, such as privatisation of the management, partial privatisation, functional privatisation and total privatisation. The basic data concerning legal regulations currently being in force is also included. Legal status is an important obstacle for the process of privatisation, since the regulations are incoherent and ambiguous. The project of Commercialisation and Privatisation of Independent Public Health Care Facilities Law was presented in 2001, but the political situation was unfavourable for such a regulation. The paper discusses also the potential profits that may be gathered during the privatisation, which may be proved by the experiences in the field of out-patient care, as well as the examples of other countries. It is stated that success of privatisation depends on changes in the structure of sources of financing health care system. The last part of the article describes the previous course of privatisation processes of in-patient sector. Presently about 18% of hospitals in Poland are private facilities, but their role in the system is marginal. The case of hospital in Wiecbork, which was the first bankrupt private hospital in Poland, is presented as the example of dangers caused by the invalid law. The final conclusion is that there is an urgent need in Poland to create clear and coherent legal frames for the privatisation of hospitals, as well as harmonisation of the changes with general reforms of the health care system. PMID- 16705879 TI - Evaluation of human exposure to single electromagnetic pulses of arbitrary shape. AB - Transient current density J(t) induced in the body of a person exposed to a single magnetic pulse of arbitrary shape or to a magnetic jump is filtered by a convolution integral containing in its kernel the frequency and phase dependence of the basic limit value adopted in a way similar to that used for reference values in the International Commission on Non-lonising Radiation Protection statement. From the obtained time-dependent dimensionless impact function W(J)(t) can immediately be determined whether the exposure to the analysed single event complies with the basic limit. For very slowly varying field, the integral kernel is extended to include the softened ICNIRP basic limit for frequencies lower than 4 Hz. PMID- 16705880 TI - Noise-induced gastric lesions: a light and scanning electron microscopy study of the alterations of the rat gastric mucosa induced by low frequency noise. AB - INTRODUCTION: Exposure to low frequency noise (LFN) can lead to vibroacoustic disease (VAD), recognized as a systemic disease with lesions in a broad spectrum of organs. Although gastrointestinal complaints are common among individuals exposed to noise, only few studies tried to evaluate the digestive lesions. The authors performed this study in order to investigate gastric lesions in an animal model of VAD. MATERIAL AND METHODS: Adult Wistar rats were exposed to continuous LFN. After five weeks they were sacrificed. The stomachs were studied by light microscopy and scanning electron microscopy, and compared with stomachs of animals kept in silence. RESULTS: Superficial erosions were present in the noise exposed animals. Massive cell death of the gastric epithelium was observed, both by light and electron microscopy. DISCUSSION: The erosions, reflecting cellular degeneration and death, occurred without inflammation, similar to what has been observed in other LFN-exposed organs. PMID- 16705881 TI - Reference frameworks for the health management of measles, breast cancer and diabetes (type II). AB - This paper presents reference frameworks which order effective and feasible policies and interventions for the health management of measles, breast cancer and diabetes (type II). These reference frameworks can be used to rapidly appraise regional health policy documents and existing health management systems. Furthermore, the reference frameworks can serve health policy makers for the planning of health management measures. PMID- 16705882 TI - Avian influenza. PMID- 16705883 TI - [Use of the comet test in occupational medicine and industrial toxicology: considerations and prospects]. AB - The comet test (Single Cell Gel Electrophoresis or SCGE) is an easy to perform, rapid and highly sensitive genotoxicity assay; it requires small amounts of biological substrate and is applicable in vivo and in vitro to a wide variety of cells and tissues. Modified versions of the comet test able to detect oxidative damage or the effects of agents inducing DNA-DNA or DNA-protein crosslinks are available. Similarly to other genotoxicity assays, the comet test is not predictive far individual cancer risk. Limitations and critical features presently linked to comet test applications, with particular regard to the biomonitoring of individuals exposed to genotoxic agents, include: lack of sensitivity with respect to aneugens (agents inducing numerical chromosomal aberrations), possible underestimation of genotoxic potency of agents with mixed action mechanisms, sensitivity depending on the genotoxic agent itself, dependence an biological substrate with regard to the influence of cytotoxicity on the assay results, influence of age, tobacco smoke, alcohol and drug consumption, diet, kinetics of DNA adducts and DNA repair mechanisms. Consequently, a routine use of the comet test in biological monitoring of individuals that are occupationally and environmentally exposed to genotoxic agents is submitted to its validation by multicentric studies on large population samples with different exposure patterns, with a suitable characterisation of the role played by the above mentioned factors. Human studies should also be increasingly focused on the direct cellular targets of exposure to genotoxicants (e.g. epithelial cells of oral cavity and airways). PMID- 16705884 TI - [The main illnesses causing absence from work. Analysis of the INPS data bank]. AB - Understanding the reasons which underlie absence from work due to illness can allow us to prevent harmful events, either in private life or professional activity, which represent a risk for the health of workers and can reduce their personal and professional capacities. OBJECTIVES: To describe the frequency of illness in a population of workers employed by the Marche region, showing: the most common pathological events, their length and the differences in occurrence linked to gender and age of the workers. MATERIALS AND METHODS: We analyzed the Marche Region INPS data bank regarding medical certificates presented by workers for the years 1999 and 2000. These medical certificates were registered by the Institute according to the nosological code (ICD-9-CM classification), length of illness, gender and age of workers. RESULTS: The incidence of illness in the specified two years decreases in the second year of observation passing from 0.67 to 0.64. The number of certificates for the two years is similar for males but increases significantly for females ((p < 0.001). In general the most frequent illnesses were infectious or parasitical ones (31%), followed by those affecting the respiratory apparatus (17%). Compared to women, men presented a higher risk for: burns, traumas and poisoning (OR 1.92 IC 1.81-2.03) and diseases of the cardio-circulatory apparatus (OR 1.75 IC 1.55-1.97). Regarding length of absence, the most common was between 4 and 7 days, even though a reversal of tendency was registered in 2000 with a decrease in short and very short absences from work in favour of average and long ones. The older workers, those over 40 years of age, tend to produce fewer certificates compared to their younger colleagues, but of longer absence. The highest number of days off work can be found among the older workers. CONCLUSIONS: According to the data we analyzed, absence from work due to illness is a complex phenomenon regarding not only gender differences but also the age of workers and the kind of work they perform. PMID- 16705885 TI - [Interactions between solvents and noise: state of the art]. AB - There are several environmental and occupational scenes where a co-exposure of noise and solvents occurs, therefore between a neurotoxic and a ototoxic agent. Epidemiological and experimental studies with simultaneous exposure to noise and solvents like toluene, styrene and carbon disulfide have been analysed. These studies investigate possible ototoxic effects through audiometry, auditory brainstem response, potential recall and reflex decay test. The current available data show that with high exposure levels, that are responsible for tissue damage by themselves, the interaction between noise and solvents could occur. The present knowledge can not permit a conclusion for low exposure levels. PMID- 16705886 TI - [Musculoskeletal disorders in piano students of a conservatory]. AB - A four parts questionnaire administered to piano students of Conservatory "T. Schipa" of Lecce, southern Italy, was used to determine the prevalence of instrument-related problems. Among 121 responders, 48 (39.6%) were considered affected according to pre-established criteria. Univariate analyses showed statistical differences for mean age, number of hours spent playing per week, interval without breaks, lack of sport practice and acceptability of "No pain, no gain" criteria in students with music-related pains compared with not affected pianists. No association with hand site was found in pianists with only upper limbs diseases. The multivariate analyses performed by logistic regression confirmed the independent association for the risk factors age, lack of sport practice and acceptability of "No pain, no gain" criteria. Differently from several studies older students were more frequently affected and no difference in the prevalence rate was found in females. Findings suggest a probable causal contribution of fixed postures in the development of PRMDs in pianists in addition to repetitive movements of upper limbs. PMID- 16705887 TI - [Recent prevention strategies and occupational risk analysis: Control Banding and Sobane]. AB - Employers are responsible for the prevention of risks and must provide for the safety and health of their workers. They are obliged to apply the general principles of prevention: to avoid, where possible, any risk; to characterize and hence to estimate residual risks; to eliminate risks at the source; to adjust jobs to the needs of workers and not workers to the jobs. When we pass to the practical performance of these shared principles we introduce many problems: problems concerning terminology; problems in estimating the nature of the risks that are faced; coordination problems between the subjects that preside over prevention; problems arising from the different typology of the companies investigated In order to answer these questions the "Industrial Hygienists" have long since created various strategies for the prevention and control of risks. Among different models the methods Control Banding and Sobane-Deparis are undoubtedly the most promising. Control Banding is designed to assist especially Small and Medium Enterprises in complying with the chemical safety regulations, the scheme uses the R phrases that in Europe must be assigned to potentially harmful chemicals by the manufacturer of the chemical. R phrases describe the most important harmful effects of a chemical and have been adopted in many non European countries also. The combination of the hazard classification of the chemical and assessment of the exposure potential will allow understanding of the level of risk thus leading the person carrying out the assessment to an appropriate control method. Occupational hygienists with experience of assessing occupational exposure to chemicals agreed parameters that could be used to give reasonable indications of exposure potential. One of them is quantity being used and three categories--small, medium and large--are defined. The likelihood of the chemical becoming airborne has been addressed by defining solids according to levels of dustiness and liquids according to volatility. A simple graph that uses the boiling point of the chemical and the process operating temperature assigns the chemical a high, medium or low volatility rating. The user now has enough information to identify the control approach required to adequately reduce exposures to the chemical Occupational hygienists agreed on three broad control approaches: General Ventilation; Engineering Control; Containment. However it is recognised that in some cases specialist advice will be needed. The user takes the hazard group, quantity and level of dustiness/volatility and matches them to a control approach using a simple table. The controls are described in control guidance sheets, which comprise both general information and, for commonly performed tasks, more specific advice. The second section of the document describes a risk-prevention strategy, called SOBANE, in four levels. These four levels are: screening, where the risk factors are detected by the workers and their management, and obvious solutions are implemented; observation, where the remaining problems are studied in more detail, one by one, and the reasons and the solutions are discussed in detail; analysis, where, when necessary, an occupational health (OH) practitioner is called upon to carry out appropriate measurements to develop specific solutions; expertise, where, in very sophisticated and rare cases, the assistance of an expert is called upon to solve a particular problem. The method for the participatory screening of the risks, Deparis, is proposed for the first level screening of the SOBANE strategy. The aim of Sobane strategy is to make risk prevention faster, more cost effective, and more effective in coordinating the contributions of the workers themselves, their management, the internal and external OH practitioners and the experts. PMID- 16705888 TI - [The Carabinieri Vice-Brigadiere Salvo D'Acquisto Gold Medal for Military Valour. Champion of justice and social solidarity]. AB - The paper recalls the life and the altruistic sacrifice that led Salvo D'Acquisto to be known and remebered as a hero. On the 23rd September 1943 the Carabinieri Vice-Brigadiere D'Acquisto saved 22 human lifes accusing himself of a presumed attack against the German occupants, although he was innocent. On the first anniversary of his execution by shooting, he was made gold medal for military valour. The cause for his beatification is currently under examination by the Holy See. PMID- 16705889 TI - [Occupational risks among public safety and security forces]. AB - The present paper tries to identify the occupational risk factors (physical, chemical, biological, psychological), variable depending on jobs and tasks, to which the heterogeneous public safety/security workers are exposed. The fight against criminality and public order maintenance imply (sometimes fatal) traumatic risks, and expose to psychophysical and sensorial tiring, unfavourable macro- and microclimatic conditions, the risk of baropathy (air navigation, underwater activities), noise (generated by firearms and several other sources), vibrations and shakings (automatic weapons, transport vehicles), the risk of electric injury, ionizing (X and gamma rays) and non-inonizing (ultraviolet rays, microwaves and radiofrequencies, electromagnetic fields) radiations. Chemical hazards include carbon monoxide and other combustion products (fires, urban traffic), substances released in chemical accidents, tear gases, lead (firing grounds, metal works, environmental pollution), solvents, lubrificants and cutting oils (mechanic repair and maintenance), laboratory materials and reagents, irritant and/or sensitizing agents contained in gloves. The main biological risks are tetanus, blood-borne diseases (viral hepatitis, AIDS), aerogenous diseases (e.g., tuberculosis, Legionnaire's disease, epidemic cerebrospinal meningitis), dog- or horse-transmitted zoonosis. Finally, emotional, psychosomatic and behavioural stress-related disorders (e.g., burn-out syndrome, post-traumatic stress disorder) are typically frequent. The presence of numerous and diversified hazards among public safety/security forces imposes the adoption of occupational medicine measures, including risk assessment, health education, technical and environmental prevention, personal protective devices, sanitary surveillance and biological monitoring, clinical interventions (diagnosis, therapy and rehabilitation of occupational accidents and illnesses), prompt medico-legal evaluation of occupational-related compensation claims. PMID- 16705890 TI - [Application of the legislation on safety occupational hygiene in the Military Navy]. AB - The paper analyses the consequences of the promulgation legislative decree 626/94 on the sanitary organization of the Armed Forces and, particularly, of the Italian Military Navy. In this context, the authors describe the current safety and occupational hygiene organization, the tasks of the vigilance organs, and the actuative modalities of preventive measures on the ships (during navigation and in shipyard). PMID- 16705891 TI - [Stress among public safety and security forces. Genetic and physiopathology]. AB - Hans Selye used the word "stress" exactly 68 years ago, in hi paper "A syndrome produced by diverse nocuous agents", published in Nature. The syndrome termed as "stress" induces hormonal autonomic responses and, over time, these hormonal changes can lead to ulcers, high blood pressure, arteriosclercosis, arthritis, kidney disease and allergic reactions. The aim of this study is to analyse the General Adaptation Syndrome (GAS), the response of the body to external stimuli, and the current knowledge about biochemistry, physiology and stress genetics. The police forces work is a complex profession, particularly liable to stress disease, as the adverse occurrences of the social life add to the stressing events specifics of callings regarding public order keeping. The results of the present study suggests the advisability of instituting a preventive monitoring for these workers. PMID- 16705892 TI - [The sanitary protection of Armed Forces employed abroad]. AB - After recalling the numerous peace expeditions of the Italian Armed Forces in foreign countries, the author underlines the several health risk factors that such missions imply. The assessment of the biological risk should be based on the knowledge of the local sanitary situation and on the analysis of the operative characteristics of the mission: prevention is based on vaccinations (with plans based on the operative tasks and destinations) and on antimalaric chemoprophylaxis, carried out following WHO indications. In conclusion, the current organization of military field hospitals is briefly described. PMID- 16705893 TI - [Ergonomy and videoterminals]. AB - After defining some concepts relative to ergonomy and videoterminals (VDT), the paper describes the main health risks for VDT workers (visual, muscle-skeletal and stress disorders). The ergonomic requirements established by the Italian legislation for VDT work are then examined, including illumination, the working environment, the working desk and seat, the video-screen, the keyboard, the upper body position, pauses and physical exercise, training and information. Recent regulations (in particular the law 422/2000) on the sanitary surveillance of VDT workers are finally reported. PMID- 16705894 TI - [Fitness for drivers]. AB - Driving vehicles for transportation of people and/or goods is a high responsibility job, that exposes to several occupational risks (psychophysical tiring; health damages by weight lifting, noise, vibrations and shakings; toxic and irritative effects by atmospheric pollutants). The Italian legislation defines the minimal psychophysical requirements for the driving licence, that is indispensable to be employed. The occupational physician should then provide the sanitary surveillance and cooperate to training and education. PMID- 16705895 TI - [Noise pathology]. AB - After defining noise and describing the physical characteristics that determine its noxious capability, the paper discusses the etiopathogenesis, clinical features and diagnostic criteria (anamnestic, audiometric, exclusion of other causes) of noise-induced damage. In conclusion, the legislative (preventive) and medico-legal issues of noise pathology are presented. PMID- 16705896 TI - [Shiftwork. Impact on health and safety in the working environment]. AB - Biological rhythms are highly disrupted by night shiftwork (NSW), and any perturbation of social and family life negatively affects performance efficiency, health and social relations. These undesirable aspects have acute and chronic components. The effects manifest themselves not only as increased accidents' frequency, but also as sleep disturbances, excessive daytime sleepiness, psychosomatic disorders that may variously interact to configure a "shift-lag" syndrome, with acute and chronic manifestation. Chronic effects increase the risk of psychoneurotic, cardiovascular and gastrointestinal diseases. The effects of NSW on women are much more pronounced because of their reproductive function and family obligations. Recent Italian legislation (1999, 2003) on night-work has essentially recognised it as a new risk factor and has established that workers' health should be safeguarded through preventive check-ups and regular controls by occupational health physicians. This involves that now occupational health physicians are required to inform workers on coping strategies, and carefully assess health disorders with absolute or relative contraindications. Data from international literature and from our group production are revised and discussed. PMID- 16705897 TI - [Issues regarding alcoholic intoxication and driving]. AB - After recalling the risks deriving from alcohol abuse at work, the paper discusses the complex problems (medical, social, juridical) related to drinking and driving. Acute intoxication may be adequately identified (also in the medico legal setting) through direct or indirect measurement of blood ethanol concentration, whereas the diagnosis of alcohol abuse and binge drinking (useful to assess fitness to work and/or driving) is complicated by the scarce efficiency of the currently available biomarkers. Alcohol abuse control and prevention among Italian Police Forces are ruled by the Ministerial Decree 30 June 2003, n. 198. PMID- 16705898 TI - [Absorption and metabolism of ethyl alcohol: diversity in two sexes]. PMID- 16705899 TI - [Use and abuse of illicit drugs, also in the light of the new Italian legislation]. AB - After presenting the new Italian legislation on the subject, the author describes the operative procedures, employed by the "Centro Ricerche di Laboratorio e di Tossicologia Forense" of the Italian State Police, for detection of illicit drugs' use by personnel on duty. The results obtained from 1998 to 2003 (on over 15,000 subjects/year) are presented. On this sample, sporadic positive (for cannabinoids) cases were found. The number of positive individuals would probably be higher, whether randomised controls were made without notice, and whether non conventional matrices (e.g. keratinised tissues) were utilized to identify previous abuse. PMID- 16705900 TI - [The risks of out of area missions: depleted uranium]. AB - Depleted uranium (DU), a waste product of uranium enrichment, has several civilian and military applications. It was used as armor-piercing ammunition in international conflicts and was claimed to contribute to health problems, known as the Gulf War Syndrome and recently as the Balkan Syndrome. Leukaemia/Limphoma cases among UN soldiers in the Balkans have been related hypothetically to exposure to DU. The investigations published in the scientific literature give no support for this hypothesis. However future follow-up is necessary for evaluation of long-term risk. PMID- 16705901 TI - [Clothing in the protection of public safety and security forces]. AB - The Ministerial Decree 19 February 1992 indicates the characteristics of the equipment and of the different uniforms adopted by the Italian Police Forces. The paper describes the occupational risks (due to physical, chemical and biological agents) and the protective equipments of particular police activities and specialties: urban and road police, air and naval units, shooting instructors, laboratory technicians (scientific police), mechanic repair and maintenance, units working with dogs or horses. PMID- 16705902 TI - [Health protection among urban police forces]. AB - After paying homage to the guest Authorities and to the organizers of the meeting, the author briefly describes the main occupational risk factors of local police forces: ergonomic and postural agents, work with videoterminals, air pollution, noise, cold-humid climate, stress. In conclusion, the symbolic importance of the uniform and the related civil and moral responsibilities are recalled. PMID- 16705903 TI - [Man, physician, scientist: in memory of Salvatore Maugeri]. PMID- 16705904 TI - [Role of Salvatore Maugeri in occupational medicine of his time]. PMID- 16705905 TI - [Between Occupational Medicine and Rehabilitation: Salvatore Maugeri, precursor of a new health model of Excellence]. PMID- 16705906 TI - Gender discrimination may be worse than you think: testing ordinal interactions in power research. AB - The authors reanalyze the data of a study by S. M. Elias and R. J. Loomis (2004), which aimed to determine how an instructor's gender may influence his or her ability to gain student compliance. S. M. Elias and R. J. Loomis observed few significant gender effects using traditional multivariate analyses of variance. The authors reanalyze this data using the more appropriate statistical techniques for detecting ordinal interactions recommended by M. J. Strube and P. Bobko (1989) and S. M. Elias (2004). An ordinal interaction occurs when 1 cell of a 2 x 2 design is responsible for a significant interaction (e.g., female instructors suffering only when rated by male students). Reanalysis of the data resulted in more robust findings. PMID- 16705907 TI - Prawn-in-a-tube procedure: habituation or associative learning in cuttlefish? AB - The prawn-in-a-tube procedure (J. B. Messenger, 1973a) has been used almost exclusively to study associative learning in cuttlefish. In two experiments, the authors sought to determine whether the decline in attack responses observed in this procedure was best accounted for by habituation or associative learning. Results of Experiment 1 revealed an asymmetrical stimulus-specificity effect that could be interpreted as either an instance of habituation or of associative learning. Results of Experiment 2 demonstrated that the response decline could not be reversed following the presentation of a dishabituatory stimulus. The combined results of these experiments support the conclusion that the response decline is best viewed as a resulting of associative processes. The authors considered whether the response decline represents extinction or passive avoidance. PMID- 16705908 TI - Gender and ethnic differences in sexual attitudes at a Hispanic-serving university. AB - The authors studied sexual attitudes in 328 university students from 10 undergraduate classes at a Hispanic-serving university near the Texas-Mexico border. Men (n = 128) and women (n = 199) rated their level of agreement with 38 items on a revised sexuality questionnaire. There were 283 self-identified Hispanic students and 44 self-identified non-Hispanic students in the sample. For the most part, these students agreed with one another, although there were statistically significant differences, with women being less permissive or more sex negative than men were on items relating to oral sex, premarital intercourse, love and sex, masturbation, Playboy magazine, and pornography. For the entire sample, 26 of the 38 items showed statistically significant gender differences. For Hispanics only, 23 of the 38 items showed statistically significant differences, with gender differences those of the sample as whole. There were 8 statistically significant differences between Hispanic and non-Hispanic students, with Hispanic students tending to be more conservative. The authors discuss findings that are contradictory to D. M. Buss's (1999, 2000) evolutionary psychology jealousy theory, namely that women in the present study were more jealous than men were on the sexual intercourse item. PMID- 16705909 TI - Effects of S+ and S- separation on gradient shifts in humans. AB - Following single stimulus training, responding during a generalization test tends to be distributed around the positive stimulus (S+). However, if participants are trained instead to discriminate the S+ from a negative stimulus (S-), the response gradient often shifts away from the S- and toward the opposite end of the stimulus continuum. In this experiment, the author examined the basis of gradient shifts with 72 college undergraduates. The research especially examined how gradient shifts are affected by the physical similarity of the S+ and the S- and by the ease with which the two stimuli can be compared. For the former manipulation, the author randomly assigned participants to either a control condition in which only the S+ was shown or a discrimination condition in which the S- was either near to or far from the S+ on the continuum. For the latter manipulation, the author randomly assigned participants to a condition in which S+ and S- presentations were separated by intervals of 1, 15, or 30 s. The results showed that marked shifts occurred when the S+ and S- were relatively similar, but temporal separations did not affect responding. PMID- 16705910 TI - Correlations among measures of executive function and positive psychological attributes in college students. AB - Executive functions are cognitive abilities that allow for more goal-oriented and autonomous behavior. The authors examined the relationship between self-rated executive functions and 3 positive personality attributes (satisfaction with life, gratitude, and forgiveness) in a nonclinical community sample. The authors used self-report scales (Executive Function Index, Heartland Forgiveness Scale "other" subscale, Satisfaction With Life Scale, and the Gratitude Questionnaire) to examine how executive functions related to the positive psychology attributes. After controlling for age, sex, and formal education, positive relationships emerged among gratitude, satisfaction, and executive function scales of motivational drive, empathy, and strategic planning. However, an inverse relationship emerged between impulse control and forgiveness, perhaps relating to cautiousness and suggesting finer distinctions between adaptive and maladaptive forms of forgiveness. These findings suggest common psychological and neurobiological substrates for executive functions and positive psychology attributes, warranting further, more direct biobehavioral research. PMID- 16705911 TI - Alcohol self-administration by rats in the presence of a tangible object. AB - The authors used the sucrose-substitution procedure to train operant self administration of a 10% alcohol solution in 8 Long-Evans rats. After they established stable responding, they began a 10-session baseline. A 10-session experimental phase followed the baseline phase. During the experimental phase, the authors placed a large glass marble in the center of the experimental chambers before self-administration sessions. The presence of the marble decreased the rats' responding and alcohol intake significantly. The authors discussed the results in terms of distraction and the effects of concurrently available reinforcers on alcohol self-administration. PMID- 16705912 TI - Affective impressions and memorability of color-form combinations. AB - In this study the authors investigate how various impressions affect the recognition of color-form pairs. They performed a preliminary study, a main study of impression rating using the semantic differential method, and a recognition experiment. On the basis of the preliminary study, the authors chose 14 appropriate pairs of adjectives and 48 color and form stimuli and used them for the main study. Factor analysis of the main study extracted three factors: Activity, Potency, and Evaluation. In the recognition test, the authors found that color-form pairs with congruent impressions in Activity or Potency were more easily recognized than pairs with incongruent impressions. In contrast, pairs with incongruent impressions in Evaluation were recognized more easily than congruent pairs. These results are discussed in relation to the assumed network representations of affective impressions that have different characteristics depending on the three factors. PMID- 16705913 TI - Face distinctiveness and delayed testing: differential effects on performance and confidence. AB - The author investigated the effect of delayed testing on participants' memory for distinctive and typical faces. Participants viewed distinctive and typical faces and were then tested for recognition immediately or after a delay of 3, 6, or 12 weeks. Consistent with prior research, analysis of measure of sensitivity (d') showed that participants performed better on distinctive rather than typical faces, and memory performance declined with longer retention intervals between study and testing. Furthermore, the superior performance on distinctive faces had vanished by the 12-week test. Contrary to d' data, however, an analysis of confidence scores indicated that participants were still significantly more confident on trials depicting distinctive faces, even with a 12-week delay between study and recognition testing. PMID- 16705914 TI - [Treatment of bone metastases]. AB - Treatment of bone metastases differs depending on whether the original cancer was diagnosed, whether it is hormone sensitive or chemosensitive, whether metastases are exclusively found in bones and thus unique, or multiple, whether they also occur in viscera and depending on the diathesis. In the last few years, treatment has benefited from a better knowledge of the efficacy of bisphosphonates, radioisotopes, percutaneous vertebroplasty and the development of new hormone therapies and chemotherapies. Given the uncertain life prognosis in the short or longer term, it remains true that cautious use of these methods is essential to reduce as far as possible the number and duration of hospitalizations. PMID- 16705915 TI - [Major advances in pediatric heart disease research]. PMID- 16705916 TI - [Epidemiology and genetics of congenital heart diseases and cardiomyopathies in children]. AB - The incidence of congenital heart defects (CHD) is close to 1% of live birth. A high proportion of CHD is associated with chromosomal anomalies. A precise assessment of the phenotype is necessary to determine which type of chromosomal anomaly has to be identified: trisomy 21 in atrio-ventricular septal defect, deletion of chromosome 22q in conotruncal defects. Genetic heterogeneity of CHD has been proven for a wide range of defects. Phenotype-genotype correlations have been established and may be helpful to orient molecular investigations. In isolated CHD, very genes have been identified so far and the risk of recurrence in a family relies mainly on a statistical approach rather than on a molecular diagnosis. Cardiomyopathies (CM) in children are mostly secondary to CHD, myocardial ischemia, drug toxicity, viral infections, and rhythm disturbances. CM associated with genetic anomalies are seen in syndromes such as the Noonan's syndrome, in dominant familial forms and in metabolic diseases. For these latter, extensive investigations are necessary as they are responsible for 15% of CM in children younger than 2 years. They are mainly represented by lysosomal storage diseases and defects in fatty acid oxidation or respiratory chain. PMID- 16705917 TI - [Prenatal diagnosis of congenital heart disease and its consequences]. AB - Echocardiography allows the prenatal diagnosis of most congenital cardiac malformations. Although the indications for this specialized ultrasound technique are multiple, suspicion of congenital heart disease at the second trimester ultrasonographic screening has become the most frequent indication. Specially trained pediatric cardiologists can confirm the diagnosis or rule it out. If the diagnosis is confirmed, then a fetal karyotype should be proposed in most cases and thorough examination of extracardiac organs, should be performed. If the prognosis is poor, French law authorizes termination of pregnancy. If the pregnancy goes on, a multidisciplinary team should organize the perinatal care according to expected neonatal difficulties. PMID- 16705918 TI - [Cardiac syncope in children]. AB - Syncope is a frequent and usually benign problem in childhood. Vasovagal syncope is the most likely etiology and is well recognized. However, syncope can herald a potentially lethal problem, so that routine evaluation including a 12-lead standard ECG should be performed in all cases. Worrying features requiring further investigations include syncope during emotion or exercise, history of familial syncope or sudden death in the young, and any abnormality on clinical exam or ECG tracings. Structural cardiac abnormalities that may cause syncope include cardiac obstructions, pulmonary hypertension, and myocardiopathy. Children with congenital heart disease who experienced syncope should always be referred to a specialist. Primary arrhythmias which are easily diagnosed on ECG are complete atrio-ventricular block and Wolff-Parkinson-White syndrome. "Channelopathies" such as the long OT syndrome and catecholaminergic ventricular tachycardia are increasingly recognized in children, carry a high risk of sudden death and deserve a complete work up, including familial screening and lifelong treatment with beta-blockers. PMID- 16705919 TI - [Main indications of cardiac surgery and interventional catheterisation in pediatric cardiac diseases]. AB - Although prospective randomized studies about congenital heart diseases in children are virtually non existent, partly due to methodological difficulties related to the scarcity and the heterogeneity of patients with these rare diseases, general agreement has been reached concerning the indications for treating, and thus approaches to the management of most congenital heart diseases do not differ significantly between centres. Advances in surgical techniques are tremendous, allowing repair of complex diseases in younger and younger children with constantly improving survival. However, long-term follow-up of these seemingly cured survivors of heart surgery within infancy is a crucial point. PMID- 16705920 TI - [Transcatheter treatment of congenital heart disease]. AB - For more than 20 years, interventional cardiac catheterization has considerably increased in the therapeutic management of simple congenital heart disease in childhood. It is possible to correct pulmonary or aortic valvar stenosis, to close a persistent shunt as patent arterial duct or atrial septal defect. Sometimes, it can replace surgical repair and can be proposed as a first line treatment. Interventional cardiac catheterisation has several advantages for the patient: no thoracotomy, no scar, shorter hospital stay, less painful, lower morbidity and reduced cost. These techniques have also benefited from miniaturization and evolution of the occluders with time. Gene therapy, tissue engineering and new imaging modality (MRI, endovascular echo) will be the future of interventional cardiac catheterization which will occupy a more important place in the treatment of congenital heart disease in children. PMID- 16705921 TI - [Adult congenital heart diseases: physicians facing new difficulties]. AB - Nowadays, 70 to 80% of the patients with congenital heart disease reach adulthood. Transition from pediatric cares to adult medicine is sometimes difficult resulting in loss of follow up, which can be deleterious in young adult with serious disease. Specialized units have been created all around the world and now in Europe. The needs for this population are specific (employment, contraception, pregnancy). Complex operated defects will need specialized assessment and follow up (tetralogy of Fallot, atrial switch for transposition of the great arteries). Arrhythmias are an important cause of admission. Native defect are sometimes revealed in adulthood (simple: asd, arterial duct, complex double discordance, Ebstein disease), or are not operable (mainly cyanotic disease). PMID- 16705923 TI - [Patient's card: the heart is an organ with a complex architecture]. PMID- 16705922 TI - [Heart diseases in children]. PMID- 16705925 TI - [Genital infections in women. Leukorrhea]. PMID- 16705926 TI - [Acute respiratory distress in the newborn and the child. Foreign bodies of the upper respiratory tract]. PMID- 16705927 TI - [Acute abdominal and lumbar pain in the adult]. PMID- 16705928 TI - [Ulcerations or erosion of the oral and/or genital mucosae]. PMID- 16705929 TI - [Osteoporosis]. PMID- 16705931 TI - [The golden age of medical radiesthesia]. PMID- 16705930 TI - [Primary and secondary liver tumours]. PMID- 16705932 TI - [Values and valorization of the tumor tissue banks]. PMID- 16705933 TI - [Valorization of the tumor tissue banks: an active plan implemented by the Institut National du Cancer]. PMID- 16705934 TI - [Biological resources centres, the initiative of OCDE]. PMID- 16705935 TI - [Conditions for using human biological samples in scientific research: legal and ethical framework]. AB - Technological progress increased our capacities in storing biological sample for longer time. The use of biological resources and related data implies that French researchers respect the legal framework including laws on bioethics, protection of individuals involved in biomedical research and related data protection. This framework aims to protect the rights of individuals involved in biomedical research and to allow the identification of biobanks by French institutions. The recent revision of the French law on bioethics offers new perspectives to broaden the use of previously collected and stored biological resources. However, the legal French framework on biological resources does not seem basically applicable to tumor cell and tissue banks with respect to the specificities of their constitution. PMID- 16705936 TI - [Contractual relationships between biobanks and researchers, the material transfer agreements]. AB - Daily interfaces actual experiences by practitioners and researchers about human biological resources for cancer are at the same time complicated and constantly evolved. PMID- 16705937 TI - [Valorization of biological resources in tumour libraries]. AB - The transfer and commercialization of biological materials, whether in the form of tumour samples, tissue samples or chemicals, and of the data base pertaining to such material have become a subject of considerable importance for both the private and public sectors involved in medical research. In order to fully appreciate and apprehend the process for the protection and the valuation of the transferred material, intellectual property law must be taken into account. As a result, a distinction is made between the tangible and intangible elements of the biological material and of the attached data base, thus providing the transferring entity the possibility to claim property rights to future intellectual property arising from the research regarding the transferred material. The transfer of biological material and attached data base without such contractual provisions can lead to the loss of this potential value as well as of physical and legal control over the material transferred by the providing entity. The intentions and the assumptions of the parties must be negotiated and written into terms of contract, at the risk of losing future value due to unexpressed assumptions concerning intangible property rights. PMID- 16705938 TI - [The CHU-CRLCC-UNSA tumour/tissue bank of the Nice area]. AB - The human tissue bank of Nice associates the CHU, the Antoine Lacassagne Center, and the University of Nice Sophia Antipolis. This bank has been labelled by the French Ministery of Health in 2003, and has been open in September 2004. Two main collections have been setting up: the tumoral and non tumoral thyroid pathology and the tumoral and non tumoral pleuro-pulmonary pathology collections. Other collections will be soon incorporated in this Biobank. The main criteria of quality connected with these collections are described. These different criteria of quality have been defined before the beginning of the collect, and thus, they have been initially strictly applied. The quality of the tissue stored at low temperature, the accuracy of the clinical and histopathological data associated with the tissue specimens, and the different procedures, should allow to obtain a labellisation by different appropriate organisms. This is crucial since the different research projects developped from human tissue collections, used expensive and accurate technologies (microarray, CGH array, proteomic, real time PCR, laser capture microdissection, etc.), and the results obtained need to be reproductible. An optimal quality both for the stored tissues and for the procedures applied in a human tissue bank is obliged since some of these tissues specimens are going to be used to try to define new prognosis criteria of tumours and new therapeutic strategies for the patients. PMID- 16705939 TI - [The consortium of Biological Resource Centres (BRC) and tumour cell and tissue banks in the Marseilles metropolitan area]. AB - The goal of this presentation is to describe current and future aspects of the operations within the consortium of Biological Resource Centres (BRC) and Tumour cell and tissue banks of the Marseilles metropolitan area. The consortium was created in year 2001, through the association of several tissue and cell banks that were operating for many years in Marseilles. Existing collections are not exclusively collections of tumour cells or tissues; however, the two tumour cell and tissue banks located at the Regional Cancer Research Centre and at the University Hospital account for a very significant proportion of the collections. Our collective work leads to the recognition and funding of the consortium by Inserm, through the "Collections 2003" grant. The consortium objectives are to define a common scientific strategy, to share professional practices in the logistics and database management of the banks, to establish a quality management program, and to build a common catalogue that describes existing biological resources. Through these efforts, the ultimate goal is to adopt rules that define BRC, as defined by the Organization for Economic Co-operation and Development (OECD). PMID- 16705940 TI - [Tumour banks network: the Spanish model]. AB - The banks of tumours exist since the pathological anatomy and cytology laboratories preserve tissue samples, in particular by paraffin inclusion. Nevertheless, the current biotechnological projections changed our needs to access to tissues and other biological samples. PMID- 16705941 TI - [Production of a common catalogue for several "Centres Regionaux de Lutte Contre le Cancer": a "Federation des Centres de Lutte Contre le Cancer" project]. AB - The "Federation des Centres de Lutte Contre le Cancer", (FNCLCC) has been undertaken 2 years ago, the set-up of an information system allowing the partition of data from tumour libraries managed by the twenty "Centres Regionaux de Lutte Contre le Cancer" (CRLCC). This organization was establihed on a secure transmission of basis informations about samples kept in tumour libraries. PMID- 16705942 TI - [Genomics and progress in oncology]. AB - Genomics and genetics are becoming pillars of clinical research, allowing certain diseases to be understood at the molecular level and leading to improved treatments. Successful genetic analyses in the clinical oncological setting depend on the quality of the samples collected. At clinical sites, attention must be paid to the steps taken prior to analysis, and to the fragility of the molecules to be tested. The development of a standard procedure specifically adapted to the identification and validation of biomarkers through genomics requires an understanding of the different technologies used at each time point, from the analysis to the data processing, and the implication of personnel at all levels: the health care centers, the biotech companies, and/or the pharmaceutical industries. PMID- 16705943 TI - [Genetic factors and colorectal cancers development: therapeutic impact]. AB - Colorectal tumorigenesis is associated with the progressive increase of epithelium dysplasia and wall invasion. These criteria are evaluated through histological staging, that enables a reliable estimation of patient prognosis, and is the best tool for therapeutic decision. Adjuvant chemotherapy is systematically proposed in case of lymph nodes and/or distant metastases (stages III and IV respectively). Its benefit in stage II tumors however remains unclear. Independently of the nature of the treatment, one third of all stage II-III tumors will metastasize. One important element to improve our tools for therapeutic decision is the identification of prognostic parameters, independent of the histological and morphological classifications. In a preliminary study, we allelotyped a series of 401 colon tumors and have shown that 5q and 8p allelic status were significantly predictive of the patients evolution. As a first approach, analysis of 47 tumors using microarray expression measures has allowed to validate the strong correlation between RNA levels and genomic status (i.e. mutation and allelic status) of known genes (APC, SMAD4, TP53, MLH1). We are now planning to characterize a series of 185 stage II-III colon tumors at both genomic and transcriptomic levels, in combination with the clinicopathological findings. Disease-free patients were followed at least 3 years after surgical resection. A tight collaboration of 5 departments of digestive oncology allowed to collect all clinical and biological resources for this project. Depending on our findings, correlations will be made between gene expression levels and somatic mutations of the coreesponding genes. Real time RT-PCR and immunohistochemical analyses will be performed on selected genes. Finally, biological mechanisms will be investigated to look for new therapeutic targets. PMID- 16705944 TI - [ENT cancers and genomics]. AB - ENT cancers in France - with the fourth rank after the breast, colon and prostate localizations - represent on important problem of public health. Survival at five years reaches a maximum in spite of undeniable therapeutic progress, in particular with the targeted treatments associated to conventional cytotoxic drugs. The studies in progress still make it possible to hope for new predictive and prognostic biological markers, but the complexity of these parameters, as well as the needs for heavy and expensive equipment to explore them, require a unification of the efforts and means. It is the major objective of the cooperative program PACOR. PMID- 16705945 TI - [AP-HM tumour tissue bank: molecular signature of gliomas]. AB - The AP-HM tissue bank is the largest one in France regarding some collections, including brain tumors. This collection was used to better characterize some gliomas. In particular because some benign gliomas such as pilocytic astrocytomas (WHO grade I) can be misclassified as malignant ones such as glioblastomas (grade IV) the first aim of our study was to find accurate diagnostic markers. This was done mainly by suppressive substractive hybridization (SSH). This study also provides a restrictive list of genes selectively involved in angiogenesis and invasion, which were highly expressed in GBM. Results were confirmed by real time quantitative RT-PCR in a large cohort of patients. In addition in order to find accurate markers which can predict GBM overall survival (OS) we selected three cohorts of GBM patients with distinctive OS (short survival < 6 months, long survival > 18 months and intermediate). Quantification of a series of markers involved in angiogenesis and invasion was done as well as cDNA array analysis. PMID- 16705946 TI - The risk of pulmonary complication after surgery for gallstone disease. AB - OBJECTIVES: To identify the risk factors of postoperative pulmonary complications (PPC) after surgery for gallstone disease. METHODS: A total of 544 patients undergoing open surgery for gallstone disease were retrospectively studied. Univariate and multivariate logistic regression were used to analyze each factor. RESULTS: There were 24 factors relating to PPC in univariate logistic analysis. And multivariate logistic analysis identified 5 independent risk factors associated with PPC, including perioperative aerosol therapy (OR = 5.906), duration of postoperative antacids therapy (OR = 1.145), duration of operation (OR = 1.011), preoperative serum albumin (OR = 0.689), postoperative analgesia (OR = 0.059), Logistic regression equation for predicting the risk of PPC was P(1) = 1/[1 + e-(- 9.803 + 1.776 x A + 0.135 x B + 0.011 x C - 0.372 x D - 2.835 x E)]. CONCLUSION: Perioperative aerosol therapy, duration of postoperative antacids therapy, duration of operation, preoperative serum albumin and postoperative analgesia were independent risk factors associated with PPC in patients undergoing surgery for gallstone disease. PMID- 16705947 TI - Sarcoidosis of the breast: a rare case report and a review. AB - Sarcoidosis is an idiopathic systemic inflammatory granulomatous disorder comprised of epithelioid and multinucleated giant cells with little necrosis. It usually invades the lungs with fibrosis and may also involve lymph nodes, skin, liver, spleen, eyes, phalangeal bones, and parotid glands. Breast involvement is extremely rare, but, when present, it could be confused with a benign or, more important, a malignant neoplasm. We have reported a case of sarcoidosis of the breast in a 51 years old woman with systemic manifestations of sarcoidosis (arthralgias and uveitis) associated with a breast mass and with a clinical history of fibrocystic mastopathy. A chest X-ray and a Computed Tomography (CT), with raised serum levels of Angiotensin Converting Enzyme (ACE), were compatible with a diagnosis of sarcoidosis. The mammography and ultrasonogram showed a solitary little nodular lesion localized in the breast. A breast cancer in a patient with sarcoidosis? To answer this question, the patient performed breast surgery under general anaesthesia and bioptic microscopic examination showed a typical sarcoid granuloma. PMID- 16705948 TI - Breast cancer in young women: case report and a review. AB - BACKGROUND: Breast cancer in patients under 40 years is uncommon. Surveillance, Epidemiology and End Results (SEER) program reveals that 75% of breast tumors occur in women age > 50 years, only 6.5% in women age < 40 years, and a mere 0.6% in women age < 30 years. Breast-conserving surgery with subsequent chemo radiotherapy has become the treatment of choice in women with breast neoplasm. CASE REPORT: Two young patients, 30 and 28 years respectively, with breast cancer. One patient with an atypical medullary breast carcinoma diagnosis, pT2 pN1 bipMx, Grade 3 Stage IIB, negative for receptors, Ki 67: 47%, cERB-2 negative; the other with an intraductal breast carcinoma, pT1c pN0 pMx, Grade 2 Stage I, negative for receptors, Ki 67: 85%, cERB-2 negative, p53 negative, Bcl-2 negative. The first patient underwent right radical mastectomy sec. Madden and axillary lymphoadenectomy in October 2001, started six cycles of adjuvant chemotherapy and radiotherapy on the right side of the chest and on axillary and supraclavicular lymph nodes area. After 2 years an ecotomography revealed small hypoechogenic nodules in the left breast. In December the patient underwent left radical mastectomy with positioning of an expander device. The histological exam revealed a not much differentiated intraductal carcinoma, pT1a N0 Mx, Stage I. After the surgical therapy, she follows another adjuvant chemotherapy. The second patient underwent left quadrantectomy with axillary limphoadenectomy in November 2004. Like the first-will follow several cycles of adjuvant chemotherapy and radiotherapy. DISCUSSION: Breast cancer in women under 40 years of age differ from breast cancer in older women in numerous clinical, pathological and biological features. The studies demonstrate that breast cancer arising in women under 40 years have a more aggressive profile than those of older patients. In both our patients family history of breast cancer was reported. That suggests a possible genetic susceptibility of these patients through BRCA1 and BRCA2 germ line mutations. Breast conservative surgery with chemio-radiotherapy is the most commonly used treatment breast cancer, expecially in consideration of the aggressiveness of the lesions. PMID- 16705949 TI - The clinical spectrum and treatment options of macrophage activation syndrome in the pediatric age. AB - Macrophage activation syndrome is a rare and potentially fatal complication of many childhood pathological settings, most frequently reported in systemic onset juvenile idiopathic arthritis. The disruption of the macrophage-lymphocyte interaction leads to uncontrolled proliferation of highly activated macrophages and T lymphocytes. The syndrome comprises a heterogeneous group of disorders featuring sepsis-like characteristics typically combined with impaired function of natural killer cells and cytotoxic T-cells, haemophagocytosis and hypercytokinemia, often resulting in fatal multiple organ failure. The clinical picture shows high grade fever, hepatosplenomegaly, pancytopenia, lymphoadenopathy, central nervous system involvement and consumptive coagulopathy. Macrophage activation syndrome is associated with high mortality: even though diagnostic criteria have been proposed, definite diagnosis can be a challenge for clinicians, especially in early phases. There is no standardized therapeutic protocol for macrophage activation syndrome, but it is widely recognized that aggressive treatment strategies might strongly influence prognosis. First line-therapy is usually represented by parenteral administration of high dose-corticosteroids, whilst cyclosporine is added in the steroid resistant cases. In this paper we provide clinical clues and summarize the most recent studies about pathophysiology and management suggestions for macrophage activation syndrome. PMID- 16705950 TI - A home-made meat-based formula for feeding atopic babies: a study in 51 children. AB - BACKGROUND: Several elimination diets have been suggested based on results of skin prick tests (SPTs) or IgE antibodies to foods, thus allowing the identification of the most common offending food(s), including CM (cow's milk), egg, peanut and wheat. But unbalanced, inappropriate dietary manipulation in infants with food allergy (FA) can have critically deleterious consequences. We have investigated the effectiveness of a home-made meat-based formula (HMMBF) (Rezza's diet) in babies with food-induced atopic dermatitis (AD), a common, disabling, chronic disease of infancy. PATIENTS AND METHODS: Rezza's diet was given for two months to 25 infants (median age 6.9 months) affected with AD and FA and the differences of body weight and AD severity score were recorded before and after the diet period. Data were analysed using the T and c2 tests. RESULTS: There was a significant improvement in both the evaluated parameters, whereas in 26 control atopic babies they remained unchanged. CONCLUSION: The results of our study indicate that the HMMBF, also based on the experience of several authors, is a useful oligoantigenic diet for the treatment of food-induced AD, and the prevention of the atopic march. PMID- 16705951 TI - Psychological stress measure in type 2 diabetes. AB - Psychological stress has been implicated as a cause of several psychosomatic disorders, but also as a factor that can unfavourably influence many diseases including diabetes mellitus. Measure of psychological stress in diabetes was performed by Psychological Stress Measure (PSM), a validated instrument, designed using 49 items drawn from descriptors generated by focus groups on stress. Clinical and psychological framework was assessed in a cohort of 100 type 2 diabetic patients (30 m, 70 f), aged 66.99 +/- 13.68 years considering disease grade, complications and level of instruction. Three other questionnaires were administered concurrently to all patients: Sickness Impact Profile (SIP), Functional Living Index (FLI) and SF-36 QOL. ANOVA statistical testing and Spearman correlation matrix were used also vs socio-cultural and clinical profile. Gender, obesity, diet compliance, smoking do not affect PSM response. Hypertensive patients and those with family history of diabetes show lower PSM scores, according to a sort of moderator effect on stress of concurrent and/or previous experience with chronic disease. Neuromuscular ailments are more prevalent in women; men vs women experience severe limitations of their working capacities and relational possibilities, with severe discomfort. In the whole, higher scores of PSM (greater stress p < 0.01) and lower scores of FLI (fair well being perception; p < 0.01) are reciprocally related inside any school instruction level. Despite the great reciprocal association of the PSM vs FLI and SIP, no significant correlation is found between PSM vs SF-36 QOL. Socio-cultural elements interfere, and particularly instruction level quantified as school grades achieved, with the manner of living their disease. Interventions on psychological distress of type 2 diabetes mellitus patients is warranted, specially in the groups with lower levels of instruction which may need an attentive strategy for achieving a satisfactory coping with this disease. PMID- 16705952 TI - Body fluid compartments in hypertension. AB - BACKGROUND AND OBJECTIVES: There is a correlation between the fluid and ionic homeostasis and blood pressure but it is not known if these body fluid changes represent the cause or rather the effect of the blood pressure rise. We have estimated the compartmental distribution of body fluids by means of the Bioimpedance Spectroscopy (BIS) analysis in a hypertensive cohort compared to control subjects. MATERIAL AND METHODS: We have enrolled 28 hypertensive patients (14 females, 14 males, mean age 47 +/- 5) and a sex- and age-matched control group of 37 healthy subjects (17 females and 20 males, mean age 45 +/- 8). They underwent anthropometric measurements, then extracellular (ECW) and intracellular water (ICW) were assessed using BIS. RESULTS: Both mean weight and BMI of hypertensive patients resulted significantly higher than of the control group (p < 0.05). We found higher ICW values in hypertensive compared to normotensive subjects. This difference was proportional to the difference of mean blood pressure values, reaching significance only as regards the stage II hypertensive subgroup (p < 0.03). DISCUSSION: Our data confirm that the blood pressure increases are associated to TBW, and caused mainly by ICW increases. The BIS, a simple, reliable, non invasive and cost effective methodical approach, estimating the distribution of body fluids, offers new possibility of the management of the hypertensive disease, to establish a more appropriate antihypertensive treatment. Moreover, the BIS, estimating the volume restoration of the different body compartments, may be helpful in evaluating the effectiveness of the pharmacological treatment. PMID- 16705953 TI - Clinical applications of radiofrequency in proctology: a review. AB - The radiofrequency scalpel is an innovative instrument which allows to cut and coagulate tissues in an atraumatic manner, conversely to the electric scalpel. The authors describe the use of radiofrequencies in proctology by making a literature review for every major proctologic disease (hemorrhoids, anal fistulas, anal fissure, sinus pilonidalis, hypertrophied anal papillae). Many techniques have been developed with radiofrequencies in hemorrhoids treatment: coagulation, ablation with plication, Milligan Morgan and Parks hemorrhoidectomy. In the treatment of anal fissures, radiofrequency subcutaneous lateral internal sphincterotomy has been described. For anal fistulas, both radiofrequency fistulotomy and fistulectomy. Finally, radiofrequency sinotomy for sinus pilonidalis and coagulation for hypertrophied anal papillae are present in literature. The analysis of the results obtained with radiofrequency surgery compared with those of the "classic" surgery for proctologic disease shows that in most of them radiosurgery facilitates, accelerates and improves the surgical procedure. PMID- 16705954 TI - Foot angiolipomas: the third case of the literature. PMID- 16705955 TI - Diagnostic and therapeutic approach to acute pulmonary embolism in an emergency department. AB - Pulmonary embolism (PE) is the obstruction of the pulmonary arteries by the dislodging and embolization of thrombotic material coming in most cases from the deep veins of the leg. PE is a relatively common disease with an estimated annual incidence up to 37 cases diagnosed per 100,000 persons it is the third cause of death in the United States. Clinical signs and symptoms are non specific and in the 70% of cases there isn't a correct diagnosis. The aim of this review is to summarize the state of the art of the diagnostic and treatment algorithms of PE in the evidence based medicine in order to minimize the "clinician gestalt" by the only guide for the early diagnosis and treatment of the disease. A correct diagnosis based on pre test probability, the use of computed tomographic pulmonary angiography, early anticoagulation/fibrinolysis started in the Emergency Department can change the natural history of the disease. In perspective, a combined approach of localyzed fibrinolysis and mechanical fragmentation could improve the overall outcome of these patients. PMID- 16705956 TI - Deepening ecological insights using contemporary statistics. PMID- 16705957 TI - Alternatives to statistical hypothesis testing in ecology: a guide to self teaching. AB - Statistical methods emphasizing formal hypothesis testing have dominated the analyses used by ecologists to gain insight from data. Here, we review alternatives to hypothesis testing including techniques for parameter estimation and model selection using likelihood and Bayesian techniques. These methods emphasize evaluation of weight of evidence for multiple hypotheses, multimodel inference, and use of prior information in analysis. We provide a tutorial for maximum likelihood estimation of model parameters and model selection using information theoretics, including a brief treatment of procedures for model comparison, model averaging, and use of data from multiple sources. We discuss the advantages of likelihood estimation, Bayesian analysis, and meta-analysis as ways to accumulate understanding across multiple studies. These statistical methods hold promise for new insight in ecology by encouraging thoughtful model building as part of inquiry, providing a unified framework for the empirical analysis of theoretical models, and by facilitating the formal accumulation of evidence bearing on fundamental questions. PMID- 16705958 TI - Statistics for correlated data: phylogenies, space, and time. AB - Here we give an introduction to the growing number of statistical techniques for analyzing data that are not independent realizations of the same sampling process -in other words, correlated data. We focus on regression problems, in which the value of a given variable depends linearly on the value of another variable. To illustrate different types of processes leading to correlated data, we analyze four simulated examples representing diverse problems arising in ecological studies. The first example is a comparison among species to determine the relationship between home-range area and body size; because species are phylogenetically related, they do not represent independent samples. The second example addresses spatial variation in net primary production and how this might be affected by soil nitrogen; because nearby locations are likely to have similar net primary productivity for reasons other than soil nitrogen, spatial correlation is likely. In the third example, we consider a time-series model to ask whether the decrease in density of a butterfly species is the result of decreases in its host-plant density; because the population density of a species in one generation is likely to affect the density in the following generation, time-series data are often correlated. The fourth example combines both spatial and temporal correlation in an experiment in which prey densities are manipulated to determine the response of predators to their food supply. For each of these examples, we use a different statistical approach for analyzing models of correlated data. Our goal is to give an overview of conceptual issues surrounding correlated data, rather than a detailed tutorial in how to apply different statistical techniques. By dispelling some of the mystery behind correlated data, we hope to encourage ecologists to learn about statistics that could be useful in their own work. Although at first encounter these techniques might seem complicated, they have the power to simplify ecological research by making more types of data and experimental designs open to statistical evaluation. PMID- 16705959 TI - Building statistical models to analyze species distributions. AB - Models of the geographic distributions of species have wide application in ecology. But the nonspatial, single-level, regression models that ecologists have often employed do not deal with problems of irregular sampling intensity or spatial dependence, and do not adequately quantify uncertainty. We show here how to build statistical models that can handle these features of spatial prediction and provide richer, more powerful inference about species niche relations, distributions, and the effects of human disturbance. We begin with a familiar generalized linear model and build in additional features, including spatial random effects and hierarchical levels. Since these models are fully specified statistical models, we show that it is possible to add complexity without sacrificing interpretability. This step-by-step approach, together with attached code that implements a simple, spatially explicit, regression model, is structured to facilitate self-teaching. All models are developed in a Bayesian framework. We assess the performance of the models by using them to predict the distributions of two plant species (Proteaceae) from South Africa's Cape Floristic Region. We demonstrate that making distribution models spatially explicit can be essential for accurately characterizing the environmental response of species, predicting their probability of occurrence, and assessing uncertainty in the model results. Adding hierarchical levels to the models has further advantages in allowing human transformation of the landscape to be taken into account, as well as additional features of the sampling process. PMID- 16705960 TI - Forecasting extinction risk with nonstationary matrix models. AB - Matrix population growth models are standard tools for forecasting population change and for managing rare species, but they are less useful for predicting extinction risk in the face of changing environmental conditions. Deterministic models provide point estimates of lambda, the finite rate of increase, as well as measures of matrix sensitivity and elasticity. Stationary matrix models can be used to estimate extinction risk in a variable environment, but they assume that the matrix elements are randomly sampled from a stationary (i.e., non-changing) distribution. Here we outline a method for using nonstationary matrix models to construct realistic forecasts of population fluctuation in changing environments. Our method requires three pieces of data: (1) field estimates of transition matrix elements, (2) experimental data on the demographic responses of populations to altered environmental conditions, and (3) forecasting data on environmental drivers. These three pieces of data are combined to generate a series of sequential transition matrices that emulate a pattern of long-term change in environmental drivers. Realistic estimates of population persistence and extinction risk can be derived from stochastic permutations of such a model. We illustrate the steps of this analysis with data from two populations of Sarracenia purpurea growing in northern New England. Sarracenia purpurea is a perennial carnivorous plant that is potentially at risk of local extinction because of increased nitrogen deposition. Long-term monitoring records or models of environmental change can be used to generate time series of driver variables under different scenarios of changing environments. Both manipulative and natural experiments can be used to construct a linking function that describes how matrix parameters change as a function of the environmental driver. This synthetic modeling approach provides quantitative estimates of extinction probability that have an explicit mechanistic basis. PMID- 16705961 TI - Analysis of neighborhood dynamics of forest ecosystems using likelihood methods and modeling. AB - Advances in computing power in the past 20 years have led to a proliferation of spatially explicit, individual-based models of population and ecosystem dynamics. In forest ecosystems, the individual-based models encapsulate an emerging theory of "neighborhood" dynamics, in which fine-scale spatial interactions regulate the demography of component tree species. The spatial distribution of component species, in turn, regulates spatial variation in a whole host of community and ecosystem properties, with subsequent feedbacks on component species. The development of these models has been facilitated by development of new methods of analysis of field data, in which critical demographic rates and ecosystem processes are analyzed in terms of the spatial distributions of neighboring trees and physical environmental factors. The analyses are based on likelihood methods and information theory, and they allow a tight linkage between the models and explicit parameterization of the models from field data. Maximum likelihood methods have a long history of use for point and interval estimation in statistics. In contrast, likelihood principles have only more gradually emerged in ecology as the foundation for an alternative to traditional hypothesis testing. The alternative framework stresses the process of identifying and selecting among competing models, or in the simplest case, among competing point estimates of a parameter of a model. There are four general steps involved in a likelihood analysis: (1) model specification, (2) parameter estimation using maximum likelihood methods, (3) model comparison, and (4) model evaluation. Our goal in this paper is to review recent developments in the use of likelihood methods and modeling for the analysis of neighborhood processes in forest ecosystems. We will focus on a single class of processes, seed dispersal and seedling dispersion, because recent papers provide compelling evidence of the potential power of the approach, and illustrate some of the statistical challenges in applying the methods. PMID- 16705962 TI - Hidden process models for animal population dynamics. AB - Hidden process models are a conceptually useful and practical way to simultaneously account for process variation in animal population dynamics and measurement errors in observations and estimates made on the population. Process variation, which can be both demographic and environmental, is modeled by linking a series of stochastic and deterministic subprocesses that characterize processes such as birth, survival, maturation, and movement. Observations of the population can be modeled as functions of true abundance with realistic probability distributions to describe observation or estimation error. Computer-intensive procedures, such as sequential Monte Carlo methods or Markov chain Monte Carlo, condition on the observed data to yield estimates of both the underlying true population abundances and the unknown population dynamics parameters. Formulation and fitting of a hidden process model are demonstrated for Sacramento River winter-run chinook salmon (Oncorhynchus tshawytsha). PMID- 16705963 TI - Model selection for geostatistical models. AB - We consider the problem of model selection for geospatial data. Spatial correlation is often ignored in the selection of explanatory variables, and this can influence model selection results. For example, the importance of particular explanatory variables may not be apparent when spatial correlation is ignored. To address this problem, we consider the Akaike Information Criterion (AIC) as applied to a geostatistical model. We offer a heuristic derivation of the AIC in this context and provide simulation results that show that using AIC for a geostatistical model is superior to the often-used traditional approach of ignoring spatial correlation in the selection of explanatory variables. These ideas are further demonstrated via a model for lizard abundance. We also apply the principle of minimum description length (MDL) to variable selection for the geostatistical model. The effect of sampling design on the selection of explanatory covariates is also explored. R software to implement the geostatistical model selection methods described in this paper is available in the Supplement. PMID- 16705964 TI - Sensitivity of ecological models to their climate drivers: statistical ensembles for forcing. AB - Global and regional numerical models for terrestrial ecosystem dynamics require fine spatial resolution and temporally complete historical climate fields as input variables. However, because climate observations are unevenly spaced and have incomplete records, such fields need to be estimated. In addition, uncertainty in these fields associated with their estimation are rarely assessed. Ecological models are usually driven with a geostatistical model's mean estimate (kriging) of these fields without accounting for this uncertainty, much less evaluating such errors in terms of their propagation in ecological simulations. We introduce a Bayesian statistical framework to model climate observations to create spatially uniform and temporally complete fields, taking into account correlation in time and space, spatial heterogeneity, lack of normality, and uncertainty about all these factors. A key benefit of the Bayesian model is that it generates uncertainty measures for the generated fields. To demonstrate this method, we reconstruct historical monthly precipitation fields (a driver for ecological models) on a fine resolution grid for a climatically heterogeneous region in the western United States. The main goal of this work is to evaluate the sensitivity of ecological models to the uncertainty associated with prediction of their climate drivers. To assess their numerical sensitivity to predicted input variables, we generate a set of ecological model simulations run using an ensemble of different versions of the reconstructed fields. We construct such an ensemble by sampling from the posterior predictive distribution of the climate field. We demonstrate that the estimated prediction error of the climate field can be very high. We evaluate the importance of such errors in ecological model experiments using an ensemble of historical precipitation time series in simulations of grassland biogeochemical dynamics with an ecological numerical model, Century. We show how uncertainty in predicted precipitation fields is propagated into ecological model results and that this propagation had different modes. Depending on output variable, the response of model dynamics to uncertainty in inputs ranged from uncertainty in outputs that matched that of inputs to those that were muted or that were biased, as well as uncertainty that was persistent in time after input errors dropped. PMID- 16705965 TI - Modeling seed dispersal distances: implications for transgenic Pinus taeda. AB - Predicting forest-tree seed dispersal across a landscape is useful for estimating gene flow from genetically engineered (GE) or transgenic trees. The question of biocontainment has yet to be resolved, although field-trial permits for transgenic forest trees are on the rise. Most current field trials in the United States occur in the Southeast where Pinus taeda L., an indigenous species, is the major timber commodity. Seed dispersal distances were simulated using a model where the major determinants were: (1) forest canopy height at seed release, (2) terminal velocity of the seeds, (3) absolute seed release, and (4) turbulent-flow statistics, all of which were measured or determined within a P. taeda plantation established from seeds collected from wild forest-tree stands at the Duke Forest near Durham, North Carolina, USA. In plantations aged 16 and 25 years our model results showed that most of the seeds fell within local-neighborhood dispersal distances, with estimates ranging from 0.05 to 0.14 km from the source. A fraction of seeds was uplifted above the forest canopy and moved via the long distance dispersal (LDD) process as far as 11.9-33.7 km. Out of 10(5) seeds produced per hectare per year, roughly 440 seeds were predicted to be uplifted by vertical eddies above the forest canopy and transported via LDD. Of these, 70 seeds/ha traveled distances in excess of 1 km from the source, a distance too great to serve as a biocontainment zone. The probability of LDD occurrence of transgenic conifer seeds at distances exceeding 1 km approached 100%. PMID- 16705966 TI - Improved estimates of net primary productivity from modis satellite data at regional and local scales. AB - We compared estimates of net primary production (NPP) from the MODIS satellite with estimates from a forest ecosystem process model (PnET-CN) and forest inventory and analysis (FIA) data for forest types of the mid-Atlantic region of the United States. The regional means were similar for the three methods and for the dominant oak-hickory forests in the region. However, MODIS underestimated NPP for less-dominant northern hardwood forests and overestimated NPP for coniferous forests. Causes of inaccurate estimates of NPP by MODIS were (1) an aggregated classification and parameterization of diverse deciduous forests in different climatic environments into a single class that averages different radiation conversion efficiencies; and (2) lack of soil water constraints on NPP for forests or areas that occur on thin or sandy, coarse-grained soil. We developed the "available soil water index" for adjusting the MODIS NPP estimates, which significantly improved NPP estimates for coniferous forests. The MODIS NPP estimates have many advantages such as globally continuous monitoring and remarkable accuracy for large scales. However, at regional or local scales, our study indicates that it is necessary to adjust estimates to specific vegetation types and soil water conditions. PMID- 16705967 TI - Application of an ecological framework linking scales based on self-thinning. AB - Barnes and Roderick developed a generic, theoretical framework for vegetation modeling across scales. Inclusion of a self-thinning mechanism connects the individual to the larger-scale population and, being based on the conservation of mass, all mass flux processes are integral to the formulation. Significantly, disturbance (both regular and stochastic) and its impact at larger scales are included in the formulation. The purpose of this paper is to illustrate how this model can be used to predict patch and ecosystem dry mass, and consequently system carbon. Examples from pine plantations and mixed forests are considered, with these applications requiring estimates of system carrying capacity and the growth rates of individual plants. The results indicate that the model is relatively simple and straightforward to apply, and its predictions compare well with the data. A significant feature of this approach is that the impact of local scale data on the dynamics of larger patch and ecosystem scales can be determined explicitly, as we show by example. Further, the general formulation has an analytic solution based on characteristics of the individual, facilitating practical and predictive application. PMID- 16705968 TI - Conversion from agriculture to grassland builds soil organic matter on decadal timescales. AB - Soil organic matter (SOM) often increases when agricultural fields are converted to perennial vegetation, yet decadal scale rates and the mechanisms that underlie SOM accumulation are not clear. We measured SOM accumulation and changes in soil properties on a replicated chronosequence of former agricultural fields in the midwestern United States that spanned 40 years after perennial-grassland establishment. Over this time period, soil organic carbon (SOC) in the top 10 cm of soil accumulated at a constant rate of 62.0 g x m(-2) x yr(-1), regardless of whether the vegetation type was dominated by C3 or C4 grasses. At this rate, SOC contents will be equivalent to unplowed native prairie sites within 55-75 years after cultivation ceased. Both labile (short turnover time) and recalcitrant (long turnover time) carbon pools increased linearly for 40 years, with recalcitrant pools increasing more rapidly than expected. This result was consistent across several different methods of measuring labile SOC. A model that investigates the mechanisms of SOM formation suggests that rapid formation of stable carbon resulted from biochemically resistant microbial products and plant material. Former agricultural soils of the Great Plains may function as carbon sinks for less than a century, although much of the carbon stored is stable. PMID- 16705969 TI - Patterns of litter disappearance in a northern hardwood forest invaded by exotic earthworms. AB - A field study was conducted to evaluate the effects of exotic earthworm invasions on the rates of leaf litter disappearance in a northern hardwood forest in southcentral New York, USA. Specifically, we assessed whether differences in litter quality and the species composition of exotic earthworm communities affected leaf litter disappearance rates. Two forest sites with contrasting communities of exotic earthworms were selected, and disappearance rates of sugar maple and red oak litter were estimated in litter boxes in adjacent earthworm free, transition, and earthworm-invaded plots within each site. After 540 days in the field, 1.7-3 times more litter remained in the reference plots than in the earthworm-invaded plots. In the earthworm-invaded plots, rates of disappearance of sugar maple litter were higher than for oak litter during the first year, but by the end of the experiment, the amount of sugar maple and oak litter remaining in the earthworm-invaded plots was identical within each site. The composition of the earthworm communities significantly affected the patterns of litter disappearance. In the site dominated by the anecic earthworm Lumbricus terrestris and the endogeic Aporrectodea tuberculata, the percentage of litter remaining after 540 days (approximately 17%) was significantly less than at the site dominated by L. rubellus and Octolasion tyrtaeum (approximately 27%). This difference may be attributed to the differences in feeding behavior of the two litter-feeding species: L. terrestris buries entire leaves in vertical burrows, whereas L. rubellus usually feeds on litter at the soil surface, leaving behind leaf petioles and veins. Our results showed that earthworms not only accelerate litter disappearance rates, but also may reduce the differences in decomposition rates that result from different litter qualities at later stages of decay. Similarly, our results indicate that earthworm effects on decomposition vary with earthworm community composition. Furthermore, because earthworm invasion can involve a predictable shift in community structure along invasion fronts or through time, the community dynamics of invasion are important in predicting the spatial and temporal effects of earthworm invasion on litter decomposition, especially at later stages of decay. PMID- 16705970 TI - Heterogeneity shapes invasion: host size and environment influence susceptibility to a nonnative pathogen. AB - Theoretical study of invasion dynamics has suggested that spatial heterogeneity should strongly influence the rate and extent of spreading organisms. However, empirical support for this prediction is scant, and the importance of understanding heterogeneity for real-world systems has remained ambiguous. This study quantified the influence of host and environmental heterogeneity on the dynamics of a 19-year disease invasion by the exotic and fatal pathogen, Phytophthora lateralis, within a stream population of its host tree, Port Orford cedar (Chamaecyparis lawsoniana). Using dendrochronology, we reconstructed the invasion history along a 1350-m length of infected stream, which serves as the only route of pathogen dispersal. Contrary to theoretical predictions, the temporal progression of the disease invasion was not related to a host's downstream spatial position, but instead was determined by two sources of heterogeneity: host size and proximity to the stream channel. These sources of heterogeneity influenced both the epidemic and endemic dynamics of this pathogen invasion. This analysis provides empirical support for the influence of heterogeneity on the invasion dynamics of a commercially important forest pathogen and highlights the need to incorporate such natural variability into both invasion theory and methods aimed at controlling future spread. PMID- 16705971 TI - Variation in species losses from islands: artifacts, extirpation rates, or pre fragmentation diversity? AB - Species are being lost from isolated reserves as predicted by ecological theory, prompting calls for larger reserves with higher species immigration rates. However, some large islands have lost a large proportion of their species, whereas some small islands have not lost any. Conservation efforts would be more efficient if the cause of such variation in the relationships among number of species lost, island size, and immigration rate were known. Observed species losses could be affected by the time since islands were isolated, species immigration rates, species extirpation rates, the pre-fragmentation diversity of the region relative to steady state, or overestimation of the pre-fragmentation diversity of islands. To test the last three hypotheses, I compared the intersection points of the island, intraprovincial, and interprovincial species area relationships of terrestrial mammals from nine archipelagos of land-bridge islands and terrestrial habitat isolates. Species losses from three archipelagos were greater than expected due to reduced immigration rates alone, although I could not resolve if this was due to increased extirpation rates or overestimation of the pre-fragmentation diversity of the islands. Analysis of six archipelagos indicates that the diversity of mammals in two regions of North America is currently below steady state, probably due to the extinction of mammals and glacial retreat during the late Pleistocene. These results have direct implications for reserve planning. When provincial diversity is below steady state, some combinations of reserve size and species immigration rate will allow reserves to maintain their pre-isolation diversity. However, the diversity of provinces relative to steady state is likely to vary, so conservation of a given proportion of a province may not always conserve the same proportion of its species. I present a new species-area relationship for islands formed by fragmentation that replaces the parameter c (fitted constant) with a rotation point. Estimation of this rotation point will allow reserve planners to separate the effects of extirpation and immigration rates on species losses from islands, identify provinces that are below steady-state diversity, and estimate the combinations of reserve size and immigration rate that will prevent loss of species from reserves. PMID- 16705972 TI - Identifying potential indicators of conservation value using natural heritage occurrence data. AB - Conservation planning based on the occurrence of rare species has been criticized as being too limited in scope to conserve biodiversity as a whole. Conversely, planning based on indicator taxa may lack sufficient focus to conserve those species in greatest need of conservation. An alternative approach is to identify a variety of species at risk that are associated with areas of conservation value, which is defined based on species-independent characteristics. We identified potential indicators of conservation value using occurrence data on species at risk and independent information on conservation value that incorporated indices of ecosystem integrity. We propose a taxonomically diverse group of indicator species that are strongly associated with areas of exceptional ecosystem integrity, to serve as a focus for further research and in planning for biodiversity conservation. We identify potential indicator species by defining a null model in which species at risk are equally associated with areas of high ecosystem integrity, then by conducting randomization tests to identify noncompliant species in the state of Michigan, USA. Areas of high ecosystem integrity are selected using criteria to flag (1) secure biotic communities with structural integrity and few exotic species, (2) natural areas subjected to expert review, (3) contiguous relict areas of forest interior, (4) contiguous areas of unmodified wetland, and (5) all these areas combined. We determine the spatial occurrence of species at risk using data from Michigan's statewide Natural Heritage database. The potential indicators include plants, insects, and birds. Their species identity and distribution of occurrences varies with the five scenarios, and together the species broadly cover the entire state. These species at risk, many of which occur throughout the Great Lakes region, may be used to identify additional areas potentially high in conservation value and to monitor their conservation. The ecological criteria and numerical methods we employ may be broadly applicable as Heritage Program databases in North America and parts of Latin America grow to become representative of species distributions. PMID- 16705973 TI - Lifting a veil on diversity: a Bayesian approach to fitting relative-abundance models. AB - Bayesian methods incorporate prior knowledge into a statistical analysis. This prior knowledge is usually restricted to assumptions regarding the form of probability distributions of the parameters of interest, leaving their values to be determined mainly through the data. Here we show how a Bayesian approach can be applied to the problem of drawing inference regarding species abundance distributions and comparing diversity indices between sites. The classic log series and the lognormal models of relative- abundance distribution are apparently quite different in form. The first is a sampling distribution while the other is a model of abundance of the underlying population. Bayesian methods help unite these two models in a common framework. Markov chain Monte Carlo simulation can be used to fit both distributions as small hierarchical models with shared common assumptions. Sampling error can be assumed to follow a Poisson distribution. Species not found in a sample, but suspected to be present in the region or community of interest, can be given zero abundance. This not only simplifies the process of model fitting, but also provides a convenient way of calculating confidence intervals for diversity indices. The method is especially useful when a comparison of species diversity between sites with different sample sizes is the key motivation behind the research. We illustrate the potential of the approach using data on fruit-feeding butterflies in southern Mexico. We conclude that, once all assumptions have been made transparent, a single data set may provide support for the belief that diversity is negatively affected by anthropogenic forest disturbance. Bayesian methods help to apply theory regarding the distribution of abundance in ecological communities to applied conservation. PMID- 16705974 TI - Elasticity analyses of size-based red and white abalone matrix models: management and conservation. AB - Prospective elasticity analyses have been used to aid in the management of fished species and the conservation of endangered species. Elasticities were examined for deterministic size-based matrix models of red abalone, Haliotis rufescens, and white abalone, H. sorenseni, to evaluate which size classes influenced population growth (lambda) the most. In the red abalone matrix, growth transitions were determined from a tag recapture study and grouped into nine size classes. In the white abalone matrix, abalone growth was determined from a laboratory study and grouped into five size classes. Survivorship was estimated from tag recapture data for red abalone using a Jolly-Seber model with size as a covariate and used for both red and white abalone. Reproduction estimates for both models used averages of the number of mature eggs produced by female red and white abalone in each size class from four-year reproduction studies. Population growth rate (lambda) was set to 1.0, and the first-year survival (larval survival through to the first size class) was estimated by iteration. Survival elasticities were higher than fecundity elasticities in both the red and white matrix models. The sizes classes with the greatest survival elasticities, and therefore the most influence on population growth in the model, were the sublegal red abalone (150-178 mm) and the largest white abalone size class (140-175 mm). For red abalone, the existing minimum legal size (178 mm) protects the size class the model suggests is critical to population growth. Implementation of education programs for novice divers coupled with renewed enforcement may serve to minimize incidental mortality of the critical size class. For white abalone, conservation efforts directed at restoring adults may have more of an impact on population growth than efforts focusing on juveniles. Our work is an example of how prospective elasticity analyses of size-structured matrix models can be used to quantitatively evaluate research priorities, fishery management strategies, and conservation options. PMID- 16705975 TI - Maximal yields from multispecies fisheries systems: rules for systems with multiple trophic levels. AB - Increasing centralization of the control of fisheries combined with increased knowledge of food-web relationships is likely to lead to attempts to maximize economic yield from entire food webs. With the exception of predator-prey systems, we lack any analysis of the nature of such yield-maximizing strategies. We use simple food-web models to investigate the nature of yield- or profit maximizing exploitation of communities including two types of three-species food webs and a variety of six-species systems with as many as five trophic levels. These models show that, for most webs, relatively few species are harvested at equilibrium and that a significant fraction of the species is lost from the web. These extinctions occur for two reasons: (1) indirect effects due to harvesting of species that had positive effects on the extinct species, and (2) intentional eradication of species that are not themselves valuable, but have negative effects on more valuable species. In most cases, the yield-maximizing harvest involves taking only species from one trophic level. In no case was an unharvested top predator part of the yield-maximizing strategy. Analyses reveal that the existence of direct density dependence in consumers has a large effect on the nature of the optimal harvest policy, typically resulting in harvest of a larger number of species. A constraint that all species must be retained in the system (a "constraint of biodiversity conservation") usually increases the number of species and trophic levels harvested at the yield-maximizing policy. The reduction in total yield caused by such a constraint is modest for most food webs but can be over 90% in some cases. Independent harvesting of species within the web can also cause extinctions but is less likely to do so. PMID- 16705976 TI - Models to compare management options for a protogynous fish. AB - Populations of gag (Mycteroperca microlepis), a hermaphroditic grouper, have experienced a dramatic shift in sex ratio over the past 25 years due to a decline in older age classes. The highly female-skewed sex ratio can be predicted as a consequence of increased fishing mortality that truncates the age distribution, and raises some concern about the overall fitness of the population. Management efforts may need to be directed toward maintenance of sex ratio as well as stock size, with evaluations of recruitment based on sex ratio or male stock size in addition to the traditional female-based stock-recruitment relationship. We used two stochastic, age-structured models to heuristically compare the effects of reducing fishing mortality on different life history stages and the relative impact of reductions in fertilization rates that may occur with highly skewed sex ratios. Our response variables included population size, sex ratio, lost egg fertility, and female spawning stock biomass. Population growth rates were highest for scenarios that reduced mortality for female gag (nearshore closure), while improved sex ratios were obtained most quickly with spawning reserves. The effect of reduced fertility through sex ratio bias was generally low but depended on the management scenario employed. Our results demonstrate the utility of evaluation of fishery management scenarios through model analysis and simulation, the synergistic interaction of life history and response to changes in mortality rates, and the importance of defining management goals. PMID- 16705977 TI - Carryover aquatic effects on survival of metamorphic frogs during pond emigration. AB - In organisms with complex life cycles, physiological stressors during early life stages may have fitness-level impacts that are delayed into later stages or habitats. We tested the hypothesis that body size and date of metamorphosis, which are highly responsive to aquatic stressors, influence post-metamorphic survival and movement patterns in the terrestrial phase of an ephemeral pond breeding frog by examining these traits in two populations of northern red-legged frogs (Rana aurora aurora). To increase variation of body size at metamorphosis, we manipulated food availability for 314 of 1045 uniquely marked tadpoles and estimated the probability that frogs survived and emigrated using concentric rings of drift fencing surrounding ponds and Bayesian capture-recapture modeling. The odds of surviving and emigrating from the ponds to the innermost drift fences, approximately 12 m, increased by factors of 2.20 (95% credibility intervals 1.39-4.23) and 2.54 (0.94-4.91) with each millimeter increase in snout vent length and decreased by factors of 0.91 (0.85-0.96) and 0.89 (0.80-1.00) with each day's delay in metamorphosis for the two ponds. The odds of surviving and moving to the next ring of fencing, 12 m to approximately 40 m from the ponds, increased by a factor of 1.20 (0.45-4.06) with each millimeter increase in size. Our results demonstrated that body size and timing of metamorphosis relate strongly to the performance of newly metamorphosed frogs during their initial transition into terrestrial habitat. Carryover effects of aquatic stressors that reduce size and delay metamorphosis may have population-level impacts that are not expressed until terrestrial stages. Since changes in both aquatic and terrestrial systems are implicated in many amphibian declines, quantifying both immediate and delayed effects of stressors on demographic rates is critical to sound management. PMID- 16705978 TI - Do top-down or bottom-up forces determine Stephanitis pyrioides abundance in urban landscapes? AB - This study examined the influence of habitat structural complexity on the collective effects of top-down and bottom-up forces on herbivore abundance in urban landscapes. The persistence and varying complexity of urban landscapes set them apart from ephemeral agroecosystems and natural habitats where the majority of studies have been conducted. Using surveys and manipulative experiments. We explicitly tested the effect of natural enemies (enemies hypothesis), host plant quality, and herbivore movement on the abundance of the specialist insect herbivore, Stephanitis pyrioides, in landscapes of varying structural complexity. This herbivore was extremely abundant in simple landscapes and rare in complex ones. Natural enemies were the major force influencing abundance of S. pyrioides across habitat types. Generalist predators, particularly the spider Anyphaena celer, were more abundant in complex landscapes. Predator abundance was related to greater abundance of alternative prey in those landscapes. Stephanitis pyrioides survival was lower in complex habitats when exposed to endemic natural enemy populations. Laboratory feeding trials confirmed the more abundant predators consumed S. pyrioides. Host plant quality was not a strong force influencing patterns of S. pyrioides abundance. When predators were excluded, adult S. pyrioides survival was greater on azaleas grown in complex habitats, in opposition to the observed pattern of abundance. Similarly, complexity did not affect S. pyrioides immigration and emigration rates. The complexity of urban landscapes affects the strength of top-down forces on herbivorous insect populations by influencing alternative prey and generalist predator abundance. It is possible that habitats can be manipulated to promote the suppressive effects of generalist predators. PMID- 16705979 TI - Assessment of prey vulnerability through analysis of wolf movements and kill sites. AB - Within predator-prey systems behavior can heavily influence spatial dynamics, and accordingly, the theoretical study of how spatial dynamics relate to stability within these systems has a rich history. However, our understanding of these behaviors in large mammalian systems is poorly developed. To address the relationship between predator selection patterns, prey density, and prey vulnerability, we quantified selection patterns for two fine-scale behaviors of a recovering wolf (Canis lupus) population in Yellowstone National Park, Wyoming, USA. Wolf spatial data were collected between November and May from 1998-1999 until 2001-2002. Over four winters, 244 aerial locations, 522 ground-based telemetry locations, 1287 km of movement data from snow tracking, and the locations of 279 wolf kill sites were recorded. There was evidence that elk (Cervus elaphus) and bison (Bison bison) densities had a weak effect on the sites where wolves traveled and made kills. Wolf movements showed a strong selection for geothermal areas, meadows, and areas near various types of habitat edges. Proximity to edge and habitat class also had a strong influence on the locations where elk were most vulnerable to predation. There was little evidence that wolf kill sites differed from the places where wolves traveled, indicating that elk vulnerability influenced where wolves selected to travel. Our results indicate that elk are more vulnerable to wolves under certain conditions and that wolves are capable of selecting for these conditions. As such, vulnerability plays a central role in predator-prey behavioral games and can potentially impact the systems to which they relate. PMID- 16705980 TI - Prey risk allocation in a grazing ecosystem. AB - Understanding the behaviorally mediated indirect effects of predators in ecosystems requires knowledge of predator-prey behavioral interactions. In predator-ungulate-plant systems, empirical research quantifying how predators affect ungulate group sizes and distribution, in the context of other influential variables, is particularly needed. The risk allocation hypothesis proposes that prey behavioral responses to predation risk depend on background frequencies of exposure to risk, and it can be used to make predictions about predator-ungulate plant interactions. We determined non-predation variables that affect elk (Cervus elaphus) group sizes and distribution on a winter range in the Greater Yellowstone Ecosystem (GYE) using logistic and log-linear regression on surveys of 513 1-km2 areas conducted over two years. Employing model selection techniques, we evaluated risk allocation and other a priori hypotheses of elk group size and distributional responses to wolf (Canis lupus) predation risk while accounting for influential non-wolf-predation variables. We found little evidence that wolves affect elk group sizes, which were strongly influenced by habitat type and hunting by humans. Following predictions from the risk allocation hypothesis, wolves likely created a more dynamic elk distribution in areas that they frequently hunted, as elk tended to move following wolf encounters in those areas. This response should dilute elk foraging pressure on plant communities in areas where they are frequently hunted by wolves. We predict that this should decrease the spatial heterogeneity of elk impacts on grasslands in areas that wolves frequently hunt. We also predict that this should decrease browsing pressure on heavily browsed woody plant stands in certain areas, which is supported by recent research in the GYE. PMID- 16705981 TI - Land use change and nitrogen enrichment of a Rocky Mountain watershed. AB - Headwater ecosystems may have a limited threshold for retaining and removing nutrients delivered by certain types of land use. Nitrogen enrichment was studied in a Rocky Mountain watershed undergoing rapid expansion of population and residential development. Study sites were located along a 30-km transect from the headwaters of the Blue River to Lake Dillon, a major source of drinking water for Denver, Colorado. Ground water in residential areas with septic systems showed high concentrations of nitrate-N (4.96 +/- 1.22 mg/L, mean +/- SE), and approximately 40% of wells contained nitrate with delta15N values in the range of wastewater. Concentrations of dissolved inorganic nitrogen (DIN) in tributaries with residential development peaked during spring snowmelt as concentrations of DIN declined to below detection limits in undeveloped tributaries. Annual export of dissolved organic nitrogen (DON) was considerably lower in residential streams, suggesting a change in forms of N with development. The seasonal delta15N of algae in residential streams was intermediate between baseline values from undeveloped streams and stream algae grown on wastewater. Between 19% and 23% of the annual N export from developed tributaries was derived from septic systems, as estimated from the delta15N of algae. This range was similar to the amount of N export above background determined independently from mass-balance estimates. From a watershed perspective, total loading of N to the Blue River catchment from septic and municipal wastewater (2 kg x ha(-1) x yr(-1)) is currently less than the amount from background atmospheric sources (3 kg x ha(-1) x yr(-1)). Nonetheless, nitrate-N concentrations exceeded limits for safe drinking water in some groundwater wells (10 mg/L), residential streams showed elevated seasonal patterns of nitrate-N concentration and ratios of DIN to total dissolved phosphorus, and seasonal minimum concentrations of nitrate-N in Lake Dillon have increased exponentially to 80 microg/L over the last decade from an initial value near zero. Results suggest that isotopic ratios in autotrophs can be used to detect and quantify increases in N enrichment associated with land use change. The biotic capacity of headwater ecosystems to assimilate increases in inorganic N from residential development may be insufficient to prevent nitrogen enrichment over considerable distances and multiple aquatic ecosystems downstream. PMID- 16705982 TI - Water management strategies against toxic Microcystis blooms in the Dutch delta. AB - To prevent flooding of the Dutch delta, former estuaries have been impounded by the building of dams and sluices. Some of these water bodies, however, experience major ecological problems. One of the problem areas is the former Volkerak estuary that was turned into a freshwater lake in 1987. From the early 1990s onward, toxic Microcystis blooms dominate the phytoplankton of the lake every summer. Two management strategies have been suggested to suppress these harmful algal blooms: flushing the lake with fresh water or reintroducing saline water into the lake. This study aims at an advance assessment of these strategies through the development of a mechanistic model of the population dynamics of Microcystis. To calibrate the model, we monitored the benthic and pelagic Microcystis populations in the lake during two years. Field samples of Microcystis were incubated in the laboratory to estimate growth and mortality rates as functions of light, temperature, and salinity. Recruitment and sedimentation rates were measured in the lake, using traps, to quantify benthic pelagic coupling of the Microcystis populations. The model predicts that flushing with fresh water will suppress Microcystis blooms when the current flushing rate is sufficiently increased. Furthermore, the inlet of saline water will suppress Microcystis blooms for salinities exceeding 14 g/L. Both management options are technically feasible. Our study illustrates that quantitative ecological knowledge can be a helpful tool guiding large-scale water management. PMID- 16705983 TI - Quantifying ultraviolet radiation mortality risk in bluegill larvae: effects of nest location. AB - Ultraviolet (UV) radiation (280-400 nm) is an increasing threat to aquatic organisms due to stratospheric ozone depletion and reductions in concentrations of dissolved organic carbon. Because fish are most vulnerable to UV during the egg and larval stages, parental spawning site selection can strongly influence mortality risk. We examined the role of nest location in determining UV-induced mortality risk for bluegill (Lepomis macrochirus) in Lake Giles, Pennsylvania, USA. In a series of five short-term incubation experiments, we found that survival of yolk sac larvae across the range of depths at which bluegill spawn was significantly lower in the presence of ambient-UV levels relative to larvae that were shielded from UV radiation. In addition, survival decreased as a function of cumulative UV exposure, as measured by the number of cyclobutane pyrimidine dimers per megabase DNA in DNA dosimeters. Although UV had the potential to significantly reduce larval survival, DNA dosimeters placed in bluegill nests concurrently with incubation experiments indicated that most nests were exposed to relatively low levels of UV. Only 19% of nests had predicted UV induced mortality greater than 25%. Consequently, current levels of UV may be an important mortality source at the level of individual nests, but not at the population level. One reason for the weak predicted effect of UV on bluegill survival is that many nests were located at depths by which much of the incident UV had been attenuated. In addition, many of the shallower nests were protected by overhanging trees or other submerged structures. It is important to note that Lake Giles is highly transparent and therefore not representative of all lakes in which bluegill are found. Nevertheless, Lake Giles is a natural system and may be representative of north temperate lakes in the future. PMID- 16705984 TI - Indirect upstream effects of dams: consequences of migratory consumer extirpation in Puerto Rico. AB - Large dams degrade the integrity of a wide variety of ecosystems, yet direct downstream effects of dams have received the most attention from ecosystem managers and researchers. We investigated indirect upstream effects of dams resulting from decimation of migratory freshwater shrimp and fish populations in Puerto Rico, USA, in both high- and low-gradient streams. In high-gradient streams above large dams, native shrimps and fishes were extremely rare, whereas similar sites without large dams had high abundances of native consumers. Losses of native fauna above dams dramatically altered their basal food resources and assemblages of invertebrate competitors and prey. Compared to pools in high gradient streams with no large dams, pool epilithon above dams had nine times more algal biomass, 20 times more fine benthic organic matter (FBOM), 65 times more fine benthic inorganic matter (FBIM), 28 times more carbon, 19 times more nitrogen, and four times more non-decapod invertebrate biomass. High-gradient riffles upstream from large dams had five times more FBIM than did undammed riffles but showed no difference in algal abundance, FBOM, or non-decapod invertebrate biomass. For epilithon of low-gradient streams, differences in basal resources between pools above large dams vs. without large dams were considerably smaller in magnitude than those observed for pools in high-gradient sites. These results match previous stream experiments in which the strength of native shrimp and fish effects increased with stream gradient. Our results demonstrate that dams can indirectly affect upstream free-flowing reaches by eliminating strong top-down effects of consumers. Migratory omnivorous shrimps and fishes occur throughout the tropics, and the consequences of their declines upstream from many tropical dams are likely to be similar to those in Puerto Rico. Thus, ecological effects of migratory fauna loss upstream from dams encompass a wider variety of species interactions and biomes than the bottom-up effects (i.e., elimination of salmonid nutrient subsidies) recognized for northern temperate systems. PMID- 16705985 TI - Trajectories of zooplankton recovery in the Little Rock Lake whole-lake acidification experiment. AB - Understanding the factors that affect biological recovery from environmental stressors such as acidification is an important challenge in ecology. Here we report on zooplankton community recovery following the experimental acidification of Little Rock Lake, Wisconsin, USA. One decade following cessation of acid additions to the northern basin of Little Rock Lake (LRL), recovery of the zooplankton community was complete. Approximately 40% of zooplankton species in the lake exhibited a recovery lag in which biological recovery to reference basin levels was delayed by 1-6 yr after pH recovered to the level at which the species originally responded. Delays in recovery such as those we observed in LRL may be attributable to "biological resistance" wherein establishment of viable populations of key acid-sensitive species following water quality improvements is prevented by other components of the community that thrived during acidification. Indeed, we observed that the recovery of species that thrived during acidification tended to precede recovery of species that declined during acidification. In addition, correspondence analysis indicated that the zooplankton community followed different pathways during acidification and recovery, suggesting that there is substantial hysteresis in zooplankton recovery from acidification. By providing an example of a relatively rapid recovery from short-term acidification, zooplankton community recovery from experimental acidification in LRL generally reinforces the positive outlook for recovery reported for other acidified lakes. PMID- 16705986 TI - Spatial scale affects community concordance among fishes, benthic macroinvertebrates, and bryophytes in streams. AB - Owing to the lack of information about the distribution patterns of many taxonomic groups, biodiversity conservation strategies commonly rely on a surrogate taxa approach for identifying areas of maximum conservation potential. Macroinvertebrates or fish are the most likely candidates for such a role in many freshwater systems. The usefulness of the surrogate taxa depends largely on community concordance, i.e., the degree of similarity in community patterns among taxonomic groups across a set of sites. We examined the effect of the spatial scale of a. study on the strength of community concordance among macroinvertebrates, bryophytes, and fish by comparing the concordance between ordinations of these groups in 101 boreal stream sites. We specifically asked if communities spanning several drainages are more concordant than those originating from a single drainage system. Our results indicate that community concordance is affected by spatial extent, being variable and generally weak at the scale of individual drainages, but strong across multiple drainage systems and ecoregions. We attribute this finding to different taxonomic groups responding to similar environmental factors and sharing a similar latitudinal gradient of community structure when viewed across large spatial scales. We also identified a "gradient of concordance," with sites contributing disproportionately to community concordance being in relatively large streams with high microhabitat variability. Overall, our results suggest that the degree of community concordance among freshwater organism groups depends critically on the spatial extent of the study, and surrogate groups at the scale of single river systems should be used with caution. PMID- 16705987 TI - Quantifying relationships between bird and butterfly community shifts and environmental change. AB - Quantifying the manner in which ecological communities respond during a time of decreasing precipitation is a first step in understanding how they will respond to longer-term climate change. Here we coupled analysis of interannual variability in remotely sensed data with analyses of bird and butterfly community changes in montane meadow communities of the Greater Yellowstone Ecosystem. Landsat satellite imagery was used to classify these meadows into six types along a hydrological gradient. The northern portion of the ecosystem, or Gallatin region, has smaller mean patch sizes separated by ridges of mountains, whereas the southern portion of the ecosystem, or Teton region, has much larger patches within the Jackson Hole valley. Both support a similar suite of butterfly and bird species. The Gallatin region showed more overall among-year variation in the normalized difference vegetation index (NDVI) when meadow types were pooled within regions, perhaps because the patch sizes are smaller on average. Bird and butterfly communities showed significant relationships relative to meadow type and NDVI. We identified several key species that are tightly associated with specific meadow types along the hydrological gradient. Comparing taxonomic groups, fewer birds showed specific habitat affinities than butterflies, perhaps because birds are responding to differences in habitat structure among meadow types and using the landscape at a coarser scale than the butterflies. Comparing regions, the Teton region showed higher predictability of community assemblages as compared to the Gallatin region. The Gallatin region exhibited more significant temporal trends with respect to butterflies. Butterfly communities in wet meadows showed a distinctive shift along the hydrological gradient during a drought period (1997-2000). These results imply that the larger Teton meadows will show more predictable (i.e., static) species-habitat associations over the long term, but that the smaller Gallatin meadows may be an area that will exhibit the effects of global climate change faster. PMID- 16705988 TI - Songbird abundance and parasitism differ between urban and rural shrublands. AB - Many studies have examined differences in avian community composition between urban and rural habitats, but few, if any, have looked at nesting success of urban shrubland birds in a replicated fashion while controlling for habitat. We tested factors affecting nest survival, parasitism by the Brown-headed Cowbird (Molothrus ater), and species abundance in shrubland habitat in rural and urban landscapes. We found no support for our hypothesis that nest survival was lower in urban landscapes, but strong support for the hypothesis that survival increased with nest height. We found strong support for our hypothesis that cowbird parasitism was greater in urban than rural landscapes; parasitism in urban sites was at least twice that of rural sites. We found strong support for an urban landscape effect on abundance for several species; Northern Cardinal (Cardinalis cardinalis) and Brown-headed Cowbirds were more abundant in urban landscapes, whereas Field Sparrow (Spizella pusilla) and Blue-winged Warbler (Vermivora pinus) were more abundant in rural sites. There was support for lower abundances of Blue-gray Gnatcatcher (Polioptila caerulea) and Indigo Bunting (Passerina cyanea) with increased housing density. For six other species, edge and trail density or vegetation parameters best explained abundance. Lower abundances and greater parasitism in habitat patches in urban landscapes are evidence that, for some species, these urban landscapes do not fulfill the same role as comparable habitats in rural landscapes. Regional bird conservation planning and local habitat management in urban landscapes may need to consider these effects in efforts to sustain bird populations at regional and local scales. PMID- 16705989 TI - Post-fledging survival of northern goshawks: the importance of prey abundance, weather, and dispersal. AB - Effective wildlife conservation strategies require an understanding of how fluctuating environmental conditions affect sensitive life stages. As part of a long-term study, we examined post-fledging and post-independence survival of 89 radio-marked juvenile Northern Goshawks (Accipiter gentilis) produced from 48 nests in northern Arizona, USA, during 1998-2001. Information-theoretic methods were used to examine within- and among-year variation in survival relative to environmental (prey abundance, weather), territory (hatching date, brood size), and individual (gender, body mass) sources of variation. The results support age- and cohort-specific differences in survival that were best explained by behaviors occurring at distinct stages of juvenile development, annual changes in the density of primary bird and mammal prey species, and gender-related differences in body mass. Survival between fledging and independence increased linearly with age and varied among annual cohorts of radio-marked juveniles from 0.81 (95% CI = 0.60-0.93) to 1.00 (95% CI = 0.95-1.00) in association with annual differences in prey density; the slope coefficient for the additive effect of prey density on survival was 1.12 (95% CI = 0.06-2.19). Survival declined to 0.71 (95% CI = 0.60 0.93) shortly after juveniles initiated dispersal (weeks 8-12 post-fledging) and moved to more open habitats at lower elevations. Survival was not closely associated with weather or territory-level parameters. A comparison of the predictions of environmental-, territory-, and individual-based models of survival demonstrated that food availability was the primary factor limiting juvenile survival. This finding indicates that forest management prescriptions designed to support abundant prey populations while providing forest structural conditions that allow goshawks to access their prey within breeding areas should benefit juvenile survival. PMID- 16705990 TI - Albatross populations in peril: a population trajectory for black-browed albatrosses at south Georgia. AB - Simulation modeling was used to reconstruct Black-browed Albatross (Diomedea melanophris) population trends. Close approximations to observed data were accomplished by annually varying survival rates, reproductive success, and probabilities of returning to breed given success in previous years. The temporal shift in annual values coincided with the start of longline fishing at South Georgia and potential changes in krill abundance. We used 23 years of demographic data from long-term studies of a breeding colony of this species at Bird Island, South Georgia, to validate our model. When we used annual parameter estimates for survival, reproductive success, and probabilities of returning to breed given success in previous years, our model trajectory closely followed the observed changes in breeding population size over time. Population growth rate was below replacement (lambda < 1) in most years and was most sensitive to changes in adult survival. This supports the recent IUCN uplisting of this species from "Vulnerable" to "Endangered." Comparison of pre-1988 and post-1988 demography (before and after the inception of a longline fishery in the breeding area) reveals a decrease in lambda from 0.963 to 0.910. A life table response experiment (LTRE) showed that this decline in lambda was caused mostly by declines in survival of adults. If 1988-1998 demographic rates are maintained, the model predicts a 98% chance of a population of fewer than 25 pairs within 78 years. For this population to recover to a status under which it could be "delisted," a 10% increase in survival of all age classes would be needed. PMID- 16705991 TI - Power quality in clinical facilities. AB - This paper presents preliminary results from a power quality audit conducted at a clinical building over the last year. Voltage and current were measured at various laboratories; it was found that the main problems for the equipment installed were voltage sags and surges. The paper examines the causes and effects of power disturbances that affect computer or any other microprocessor-based equipment and analyzes the auto-protection capabilities of modern power supplies. The convenience of "enhanced power supply" or "low-cost customer-side" protection solutions is also discussed. Finally it is addresses the role of the Standards on the protection of electronic equipment and the implications for the final costumer. PMID- 16705992 TI - A tool for improving the Web accessibility of visually handicapped persons. AB - Abstract Much has been written concerning the difficulties faced by visually handicapped persons when they access the internet. To solve some of the problems and to make web pages more accessible, we developed a tool we call the "Easy Bar," which works as a toolbar on the web browser. The functions of the Easy Bar are to change the size of web texts and images, to adjust the color, and to clear cached data that is automatically saved by the web browser. These functions are executed with ease by clicking buttons and operating a pull-down list. Since the icons built into Easy Bar are quite large, it is not necessary for the user to deal with delicate operations. The functions of Easy Bar run on any web page without increasing the processing time. For the visually handicapped, Easy Bar would contribute greatly to improved web accessibility to medical information. PMID- 16705993 TI - A snore extraction method from mixed sound for a mobile snore recorder. AB - This paper presents a snore recorder that can separate snores from their delayed mixtures. This is useful to study the snore sounds of individuals when these sounds occur in a normal in-home sleeping environment, where two people are sleeping together and both produce sounds. Based on methods for blind source separation, we give a snore separator that solves the blind delayed source separation problem and provide a performance index to monitor its convergence. The separated snores can be analyzed to detect symptoms of sleep apnea prior to polysomnography or as a monitoring device after polysomnography has been performed. Experimental results show good performance of the snore separator. PMID- 16705994 TI - The effectiveness of the installation of a mobile voice communication system in a university hospital. AB - In large hospitals, collaborative clinical practice is currently emphasized, with members of various departments expected to work as a team. The importance of accurate communication among the team members is of utmost importance. To improve such communication, the introduction of mobile voice communication systems has received much attention in Japan. Shimane University Hospital also introduced a Personal Handy-phone System (PHS) for doctors. In the traditional setting, much time was wasted searching for doctors through multiple calls on fixed-line telephones. In order to measure the effectiveness of our system, the change in the number of calls made on fixed-line telephones before and after PHS installation was compared. The total number of calls was reduced by more than 35%, and the number of calls to the wards on weekdays was reduced by half. Mobile telecommunication systems with small output power, such as PHS, are known to cause little interference with medical devices which makes it possible to use mobile voice communication safely in hospitals. The improvement in communication by this systems resulted in an improvement in labor efficiency. PMID- 16705995 TI - Enabling collaborative medical diagnosis over the Internet via peer-to-peer distribution of electronic health records. AB - Recent developments in networking and computing technologies and the expansion of the electronic health record system have enabled the possibility of online collaboration between geographically distributed medical personnel. In this context, the paper presents a Web-based application, which implements a collaborative working environment for physicians by enabling the peer-to-peer exchange of electronic health records. The paper treats technological issues such as Video, Audio and Message Communication, Workspace Management, Distributed Medical Data Management and exchange, while it emphasizes on the Security issues arisen, due to the sensitive and private nature of the medical information. In the paper, we present initial results from the system in practice and measurements regarding transmission times and bandwidth requirements. A wavelet based image compression scheme is also introduced for reducing network delays. A number of physicians were asked to use the platform for testing purposes and for measuring user acceptance. The system was considered by them to be very useful, as they found that the platform simulated very well the personal contact between them and their colleagues during medical meetings. PMID- 16705996 TI - Assessing domains of uncertainty in critical Web-based healthcare communities. AB - Search engines and Web-based directories play a central role in facilitating access to online health information. In this environment consumers often find and act upon health information of variable quality, with little input from health professionals. As an added concern, when consumers access health information on the Web via a search engine they often fail to discuss the information with a healthcare provider, especially in areas of perceived medical uncertainty. For many health consumers, the Internet may be the primary (or even sole) source of health information, especially where consumers lack access to, or trust in, trained providers. To date the corresponding role of search engine technology use and efficacy has received relatively little attention, however. This study serves as an exploratory technology assessment that explains the application of keyword effectiveness indexing (KEI) analysis in estimating the ability of commercial search engines to return relevant results for healthcare consumers. Findings suggest that KEI can serve as a useful health search profiling tool, though its applicability may vary across domains. PMID- 16705997 TI - Enhancing the utility of prostascint SPECT scans for patient management. AB - This project investigated reducing the artifact content of In-ill ProstaScint SPECT scans for use in treatment planning and management. Forty-one patients who had undergone CT or MRI scans and simultaneous Tc-99m RBC/In-111 ProstaScint SPECT scans were included. SPECT volume sets, reconstructed using Ordered Set Expectation Maximum (OS-EM) were compared against those reconstructed with standard Filtered Back projection (FBP). Bladder activity in Tc-99m scans was suppressed within an ellipsoidal volume. Tc-99m voxel values were subtracted from the corresponding In-111 after scaling based on peak activity within the descending aorta. The SPECT volume data sets were merged with the CT or MRI scans before and after processing. Volume merging, based both on visual assessment and statistical evaluation, was not affected. Thus iterative reconstruction together with bladder suppression and blood pool subtraction may improve the interpretation and utility of ProstaScint SPECT scans for patient management. PMID- 16705998 TI - Model-based compression for 3D medical images stored in the DICOM format. AB - Many medical examinations involve acquisition of a large series of slice images for 3D reconstruction of the organ of interest. With the paperless hospital concept and telemedicine, there is very heavy utilization of limited electronic storage and transmission bandwidth. This paper proposes model-based compression to reduce the load on such resources, as well as aid diagnosis through the 3D reconstruction of the structures of interest, for images acquired by various modalities, such as MRI, Ultrasound, CT, PET etc. and stored in the DICOM file format. An example implementation for the biliary track in MRCP images is illustrated in the paper. Significant compression gains may be derived from the proposed method, and a suitable mixture of the models and raw images would enhance the patient medical history archives as the models may be stored in the DICOM file format used in most medical archiving systems. PMID- 16705999 TI - Access to safe drinking water: more important than health promotion. PMID- 16706000 TI - Evaluation of a Dutch public-private partnership to promote healthier diet. AB - PURPOSE: Public-private partnerships may help to promote healthy diets. We assessed customers' exposure to and the acceptability of a Dutch public-private healthy diet campaign in butcher's shops and investigated the effects on the purchase of lean meat and the use of liquid cooking margarine and potential behavioral determinants. METHODS: The pretest-posttest control group design included 486 customers (242 experimental and 244 control) of butcher shops representing 64% of the original sample. Campaign exposure, acceptability, and behavioral effects were measured by a questionnaire. RESULTS: Seventy-one percent of the customers noticed the campaign. Scores on the acceptability were positive to very positive. Regression analysis revealed that customers in the experimental condition evaluated the campaign better (B = .415; p < .05) and felt more encouraged to buy lean meat (B = .252; p < .05) than customers in the control condition. No effects on behavior were found. DISCUSSION: Study design limitations included possible campaign exposure of control group participants. The study shows the feasibility and acceptability of a joint health-promoting activity through a public-private partnership, but there were no effects on behavior. PMID- 16706001 TI - Assessing the structure of smoking cessation care in the Veterans Health Administration. AB - PURPOSE: National smoking cessation practice guidelines offer recommendations regarding the processes and structure of care. Facilities routinely measure the processes of care but not the structure of care. This pilot study assessed the structure of smoking cessation care at Veterans Health Administration facilities. METHODS: Key informants at 18 sites completed a brief checklist survey adapted from national smoking cessation guidelines. Responses were compared with detailed site surveys. RESULTS: Guideline adherence was seen in identifying smokers and treating inpatient smokers. Areas of low adherence include offering incentives and defining staff responsibilities. The checklist survey showed poor correspondence with the detailed survey, with low agreement on systematic screening (kappa = .21) and higher agreement on primary care prescribing authority (kappa = .53). DISCUSSION: This pilot survey provides a potential rapid method for assessing adherence to systems recommendations from the national smoking cessation guidelines. The relatively low agreement with a more detailed survey suggests that the two surveys may have been measuring different aspects of smoking cessation care. PMID- 16706002 TI - Cigarette smoking and smoking cessation among persons with chronic obstructive pulmonary disease. AB - PURPOSE: To identify factors predictive of smoking cessation among adults with chronic obstructive pulmonary disease (COPD). Data from the 1997 to 2002 National Health Interview Surveys were analyzed for adults at least 25 years of age with COPD using logistic regression. RESULTS: Of the adults with COPD, 36.2% were current smokers. Of the current smokers and former smokers who had quit smoking during the past year, 22.9% reported not receiving cessation advice from a health care professional during the past year Although half of smokers with COPD had attempted to quit during the past year, only 14.6% were successful. Attempting to quit was negatively associated with heavy drinking but positively associated with being younger and having cardiovascular diseases, lung cancer, and activity limitation due to lung problems. Factors predictive of successful cessation included being at least 65 years old, not being poor, and activity limitation due to lung problems. CONCLUSION: This study underscores the importance of continuing to develop smoking cessation strategies for COPD patients and implementing clinical guidelines on smoking cessation among health care providers. PMID- 16706003 TI - Negative affect and risk for human immunodeficiency virus: one size may not fit all. AB - PURPOSE: Research examining the relationship between human immunodeficiency virus (HIV)-related risk behavior and negative affective states has netted mixed findings. The present study examined the potential moderating role of gender and sexual orientation on negative affect and HIV-related risk behavior. DESIGN: Measures of depression, anxiety, and HIV-related risk behavior were collected from participants at the time of pretest counseling. Analysis of variance was used to examine HIV-related risk behavior as a function of negative affect levels and gender and behavioral sexual orientation. SETTING: The study was conducted through a community-based, anonymous HIV counseling and testing site. SUBJECTS: Participants included 185 consecutively recruited men and women presenting for HIV testing. MEASURES: Participants completed a structured interview regarding HIV-related risk behavior, the Center for Epidemiological Studies Depressed Mood Scale, and the Beck Anxiety Inventory. RESULTS: Greater HlV-related risk behavior was associated with high levels of anxiety and moderate or high levels of depression for bisexual women. Greater HIV-related risk behavior was also associated with low levels of anxiety and mild to moderate levels of depression for bisexual men. CONCLUSIONS: The relationship between negative affective states and HIV-related risk behavior may vary as a function of gender and behavioral sexual orientation. Better understanding of the relationship between negative affect and risk behavior may be achieved through consideration of these sources of variation. PMID- 16706004 TI - Status of U.S. military retirees and their spouses toward achieving Healthy People 2010 objectives. AB - PURPOSE: To assess the status of U.S. military retirees and their spouses 38 to 64 years of age relative to select Healthy People 2010 objectives and to identify sociodemographic characteristics associated with select health behaviors. DESIGN: Cross-sectional analyses with self-reported standardized measures from the U.S. Department of Defense Population Health Survey, 2003. SETTING: The continental United States. SUBJECTS: U.S. military retirees and their spouses 38 to 64 years of age. MEASURES: Data for this study were self-reported responses to the Population Health Survey. Dichotomous variables were created to indicate whether each Healthy People 2010 objective had been met. Each objective was measurable with the survey by using the definitions set forth in Healthy People 2010. These objectives included healthy weight and obesity based on body mass index (height and weight); daily fruit, vegetable, and grain-product consumption; physical inactivity, moderate physical activity, and vigorous physical activity; binge drinking; cigarette use; and smoking-cessation attempts. RESULTS: The study population did not meet any of the Healthy People 2010 objectives included in this study. Sociodemographic characteristics that were associated with this result included being male, not having a college degree, and a less-than excellent self-reported general health status. CONCLUSION: Health-promotion interventions are needed to improve the health status of this population and to achieve the goals set forth in Healthy People 2010. PMID- 16706005 TI - Application of the transtheoretical model to fruit and vegetable consumption among economically disadvantaged African-American adolescents: preliminary findings. AB - PURPOSE: To examine the application of the transtheoretical model (TTM) to fruit and vegetable consumption among economically disadvantaged African-American adolescents. METHODS: Scales for measuring decisional balance, situational self efficacy, and processes of change for fruit and vegetable consumption were developed and pretested with 57 economically disadvantaged African-American adolescents. The scales and measures for assessing stages of change, demographic variables, and fruit and vegetable consumption were administered to a separate sample of 262 participants. t-tests for independent samples and analysis of variance were used to examine differences in TTM variables and fruit and vegetable consumption across stages of change. RESULTS: Two-factor solutions for decisional balance and processes of change and a three-factor solution for situational self-efficacy provided the best fit to the data. Alpha coefficients of reliability for the scales ranged from .77 (experiential change processes) to .91 (pros). Participants in action-maintenance stages evidenced higher pros, self efficacy, and fruit and vegetable consumption and significantly lower cons than did participants in precontemplation and contemplation-preparation stages. Also, participants in action-maintenance stages used processes of change more frequently than did those in precontemplation-contemplation-preparation stages. The use of experiential and behavioral processes within these stages did not differ significantly, as posited. DISCUSSION: Observed differences in TTM variables and fruit and vegetable consumption by stage of change in this sample of economically disadvantaged African-American adolescents were consistent with theory and previous applications of the model to fruit and vegetable consumption in adults. With replication studies, the TTM may be appropriate for designing interventions to increase fruit and vegetable consumption among this population. PMID- 16706006 TI - Sun care advertising in popular U.S. magazines. AB - PURPOSE: We assessed the placement of magazine advertising for sun care products to lay the groundwork for broader promotion to more diverse and high-risk demographic groups. METHODS: We reviewed 579 issues of 24 magazines published between the months of May and September from 1997 to 2002. We conducted a cover to-cover review of top-selling magazines for men, women, teens, parents, travelers, and outdoor recreation users. We determined if there were any advertisements for the following sun care products: sun tanning lotions containing sun protection factor (SPF), sunless tanners without SPF, sunscreen with SPF, moisturizers with SPF, or cosmetics with SPF (which include sunless tanners containing SPF. RESULTS: Sun care products, including sunscreens, were advertised primarily in women's magazines (77%). Nearly two thirds of all sun care products advertised were either for cosmetics (38%) or moisturizers (26%) containing SPF, followed by ads for sunscreen sold as a stand-alone product (19%). None of the ads contained all of the recommendations for safe use of sunscreen: a minimum SPF of 15, both UVA and UVB protection, reapplication instructions, and an adequate application coverage of 2 milligrams per square centimeter. DISCUSSION: Magazine advertising to men, travelers, outdoor recreation users, and parents/families (totaling a circulation of 41 million readers) during this six-year period were far fewer than those for women, despite high rates of excessive sun exposure in these groups. PMID- 16706007 TI - The association of two productivity measures with health risks and medical conditions in an Australian employee population. AB - PURPOSE: To investigate the impact of health on job performance using two measures of productivity loss: (1) a self-reported measure of health-related presenteeism and (2) an objective measure of absenteeism. DESIGN: A cross sectional survey using a Health Risk Appraisal (HRA) to evaluate self-reported presenteeism and the prevalence of 12 health risks and eight medical conditions. SETTING AND SUBJECTS: Employees (n=224) of a private insurance provider in Australia. MEASURES: A Health Risk Appraisal (HRA) questionnaire was used to evaluate self-reported presenteeism on different aspects of job demands and to assess the prevalence of 12 health risks and eight medical conditions. Illness absent hours were obtained from company administrative records. RESULTS: Increased presenteeism was significantly associated with high stress, life dissatisfaction, and back pain, while increased illness absenteeism was significantly associated with overweight, poor perception of health, and diabetes. Excess presenteeism associated with excess health risks (productivity loss among those with medium- or high-risk status compared to those with low-risk status) was independently calculated at 19.0% for presenteeism and 12.8% for illness absenteeism. CONCLUSIONS: This study demonstrates an association between health metrics and self-reported work impairment (presenteeism) and measured absenteeism. The study provides a first indication of the potential benefits of health promotion programming to Australian employees in improving health and to the corporation in minimizing health-related productivity loss. PMID- 16706008 TI - Best practices for an integrated population health management (PHM) program. AB - Large employers are beginning to create much more robust and potentially effective systems of integrated health management interventions. This organization-wide activity is referred to as a "Population Health Management" (PHM) approach. Best practices are also beginning to emerge in this area, and a number of technical developments are helping shape the patterns of response from innovative employers. This initiative is rapidly emerging as a critical part of overall business and health care strategy. PMID- 16706009 TI - The top 10 causes of unprofessional conduct. PMID- 16706010 TI - Review of decisions of Professional Conduct and Appeals Committees. PMID- 16706011 TI - A zero tolerance abuse policy--what does an RN need to consider? AB - CARNA's Nursing Practice Standards and the CNA Code of Ethics for Registered Nurses support RNs in their practice and provide guidance when questions about practice arise, facilitate problem-solving and provide a focus on the values, beliefs and responsibilities that RNs have when providing nursing care, even at those times when it may seem that a patient may not want their help. PMID- 16706012 TI - [Advances in studies on chemical constituents and bioactivities of plants from flacourtiaceae]. AB - In this article, the research of chemical constituents and bioactivities in recent ten years has been reviewed of plants from the 12 genera in Flacourtiaceae related to the medicinal resources in China. The research in China about the plants from Flacourtiaceae was done very little, but many literatures have been reported abroad. The plants from Flacourtiaceae mostly contain the constituents such as aromatic glucosides, lignanoid glucosides, diterpenoids and cyclopentenoid cyanohydrin glucosides et al. These compounds or plant extracts mainly show antibacterial, antiviral, antitumor, hypolipidemic and hypoglycemic activities. The research of plants from Carrierea, Itoa and Bennettiodendron of Flacourtiaceae in China is still blank. The systemic research about chemical constituents and bioactivities of plants from these genera will play important roles in the discovery of novel natural products and active constituents, and provide valuable reference for the classifying of plants from these genera. PMID- 16706013 TI - [Microencapsulation of Chinese herb activities]. AB - Microencapsulation of Chinese herb activities was summarized, including Chinese herb microencapsulation purpose, significance, study status quo and prospecting application foreground. Microencapsulation of Chinese herb activities play a important role in Chinese herb preparation industry and modernization. With the technology developing, microencapsulation of Chinese herb activities will have widely application foreground. PMID- 16706014 TI - [Study on dynamic growth of Codonopsis pilosula]. AB - OBJECTIVE: To study on the dynamic changes of Codnopsis pilosula for the guidance of the field management. METHOD: Using the random method the influences of the cultivating density and fertilizing weight at four different levels was observed. RESULT AND CONCLUSION: The dynamic changes of the biomass, growth period andbiological changes at different growth stages were found out. The most predominant combination of the two factors, the 1.05 million roots/hm2 of the cultivating density and 240 kg x hm(-2) of the fertilizing weight reached the highest accumulation of stems & leaves; 652 mg x d(-1) the 0.6 million roots/hm2 and 240 kg x hm(-2) reached the heaviest single fresh root. The results could be used for the cultivating, fertilizing and practicing SOP of C. pilosula. PMID- 16706015 TI - [Determination of acetylharpagide in Ajuga decumbens by HPLC]. AB - OBJECTIVE: To establish a method of the quantitative determination of acetylharpagide in Ajuga decumbens. METHOD: The chromatographic conditions were as follows: a Phenomenex Luna C18 column was used, the mobile phase was composed of acetontrile-water (15:85), the flow rate was 1.0 mL x min(-1) and the UV absorbance detection was set at 197 nm. RESULT: Linearity of acetylharpagide was in the range of 0.6-3.6 microg (r = 0.9993), and the average recovery and RSD were 99.13% and 2.48%, respectively. CONCLUSION: The contents of acetylharpagide ranged 0.40%-6.39% in A. decumbens. The method was simple, accurate and sensitive. PMID- 16706016 TI - [Studies on population difference of Dendrobium officinale II establishment and optimization of the method of ISSR fingerprinting marker]. AB - OBJECTIVE: To establish and optimize ISSR-PCR system of Dendrobiwn officinale according to the ISSR-PCR characters of D. officinale. METHOD: The effects of ISSR-PCRs was examined by selecting primers and designing different concentrations of the factors in the ISSR-PCRs, the reliable ISSR-PCR systems for D. officinale populations researching was established by analyzing the reasons for occurrence of differential bands and optimizing reaction conditions. RESULT AND CONCLUSION: The optimal ISSR-PCR system in D. officinale was reported for the first time, and 25 15327012 microL ISSR-PCR system contained 10 x Taq buffer, 1.5 U Taq DNA polymerase, 1.2-1.8 mmol x L(-1) MgCl2, 80 micromol x L(-1) dNTP, 0.2 micro mol x L(-1) primer and 20 ng template DNA. The appropriate annealing temperature was among 52-60 degrees C. ISSR PCRs were significantly influenced by Taq DNA polymerase, template DNA quantity and annealing temperature, etc. The ISSR-PCR systems, which were established in this paper for studying D. officinale, could provide clear reliable abundant polymorphisms molecular markers and were proved suitable for studying population authentication and population molecular ecology of D. officinale. PMID- 16706017 TI - [The research of the adsorption and desorption behaveior of macroporous resin for glycyrrhizic acid]. AB - OBJECTIVE: To investigate the performance of static and dynamic adsorption and desorption of the three macroporous resins of AB-8, D-101 and H-103. METHOD: Compare the adsorption and desorption of the three macroporous resins for glycyrrhizic acid by determining the static adsorption, static desorption, dynamic adsorption and dynamic desorption through detecting the glycyrrhizic acid content by HPLC method. RESULT: The saturation of AB-8 is 0.75 g (crude drug)/mL (macroporous resins), the best elution reagent is 50% EtoH and it's eluation ration is 68.81% . The saturation of D-101 is 0.75 g (crude drug)/mL (macroporous resins), the best elution reagent is 60%EtoH and it's eluation ration is 64.67%. The saturation of H-103 is 0.5 g (crude drug)/mL (macroporous resins), the best elution reagent is 50%E-toH and it's elution ration is 51.18%. CONCLUSION: AB-8 and D-101 are the superior absorbents for glycyrrhizic acid. PMID- 16706018 TI - [Analysis of the volatile components of YL2000 decoction by GC-MS]. AB - OBJECTIVE: To analyze the volatile components of YL2000 decoction, which contain four herbs: rhizome and root of the Notopterygium incisum, the root of the Angelica pubescens f. biserrata, Scutellaria baicalensis and Coptis chinensis and investigate the changes of volatile constituents from those four herbs before and after compatibility of the herbal medicines. METHOD: The volatile components of YL2000 decoction were extracted by water-steam distillation, separated and identified by GC-MS. The relative percent content of each volatile component was quantified by area normalization method. The volatile components of YL2000 decoction were compared with the composition of the volatile oil from individual herb respectively. RESULT: 39 of the 146 separated constituents in volatile oil of YL2000 decoction, accounting for 85.66%, were identified and quantified. After compatibility of the herbal medicines, most volatile oil components reported by high proportion in individual herb were not detected, in the mean time, some components in volatile oil of YL2000 decoction have never been reported before in those of all four herbs. CONCLUSION: The changes of volatile oil from those four herbs before and after compatibility of the herbal medicines indicate that solubilization, chemical reactions and evaporation of some volatile components during decocting may induce changes of several components. PMID- 16706019 TI - [Study on HPLC fingerprint of different growing areas on fructus aurantii fried with bran]. AB - OBJECTIVE: To establish a method for HPLC fingerprint determination of different growing areas on Fructus Aurantii fried with bran. METHOD: HPLC chromatographic experiment was carried out by ultrasonic distilment of acetacetic ester, gradient elution mode and the detection wavelength of 320 nm. The column temperature was set at 30 degrees C, and flow rate is 1.0 mL x min(-1). Naringin served as the reference standard. Ten batches of Fructus Aurantii fried with bran in the different places were analyzed. Similarity evaluation system for chromatographic fingerprint of TCM proceeded the evaluation, The correlation coefficients between ten batches were used to define the similarity. RESULT: There were eleven characteristic peaks in Fructus Aurantii fried with bran by HPLC fingerprint which constituted fingerprint characters. Both the reproducibility and stability on HPLC fingerprint were proper. CONCLUSION: The method was accurate and simple. It can be applied to the analysis on Fructus Aurantii fried with bran. The fingerprint spectrum can be used to distinguish Fructus Aurantii fried with bran and to be its quality control. PMID- 16706020 TI - [Studies on chemical constituents in stem of Dendrobium chrysotoxum]. AB - OBJECTIVE: To investigate the chemical constituents of Dendrobium chrysotoxum. METHOD: The chemical constituents were isolated by various column chromatographic methods and structurally elucidated by spectral evidences. RESULT: Ten compounds were obtained and identified as (+)-syringare sinol (1), 5alpha, 8alpha-epidioxy 24( R)-methycholesta-6, 22-dien-3beta-ol (2), trans-3-(4-hydroxy-3-methoxyphenyl) acrylic acid octacosyl ester (3), defusin (4), 3, 4-dihydroxy benzoic acid (5), 3, 4-dimethoxy-benzoic acid (6), vanillic acid (7), 3, 4-dimethoxy-benzoic acid methyl ester (8), 3, 5-dibromo-2-aminobenzaldehyde (9), heptadecanoic acid 2, 3 dihydroxy-propyl ester (10). CONCLUSION: Compounds 1, 2 and 6-10 were isolated from this plant for the first time. PMID- 16706021 TI - [Studies on the chemical constituents from stem of Chirita longgangensis var. Hongyao]. AB - OBJECTIVE: To isolate and elucidate the constituents from stem of Chirita longgangensis var. hongyao. METHOD: The constituents were extracted with methanol and isolated by chromatography on silica gel, Sephadex LH-20 and ODS. The structures were determined by NMR and MS spectral analysis. RESULT: Seven compounds were identified as 2-hydroxy-7-methyl-9, 10-anthraquinone (1), 2-methyl 9, 10-anthraquinone (tectoquinone) (2), ursolic acid (3), vanillic acid (4), beta sitosteryl-D-glucoside-6'-palmitate (5), beta-sitosterol (6), daucosterol (7), respectively. CONCLUSION: Compounds 1, 2, 3 and 5 were isolated for the first time from the family Gesneriaceae, compounds 4 and 6 were isolated for the first time from the genus Chirita, and compound 7 was isolated for the frist time from Chirita longgangensis var. PMID- 16706022 TI - [Studies on chemical constituents in roots of Heracleum rapula]. AB - OBJECTIVE: To study the chemical constituents in roots of Heracleum rapula. METHOD: The constituents were isolated by column chromatography on silica gel and ODS, and identified by spectroscopic methods. RESULT: Nine compounds, osthol ( I ), bergapten (II), xanthotoxin (III), isopimpinellin (IV), imperatorin (V), isoimperatorin (VI), cnidilin (VII), phellopterin (VIII), rivulobirin A (IX) were isolated and identified. CONCLUSION: Compounds VI, VII, VIII, IX were isolated from this plant for the first time. PMID- 16706023 TI - [Effect of mica granule on the expression of gene-protein associated with cancer in gastric mucosa tissue of chronic atrophic gastritis rats]. AB - OBJECTIVE: To research the regulative effect of mica monomer granule preparation on the expression of gene associated with cancer in gastric mucosa tissue of experimental chronic atrophic gastritis (CAG) rats. METHOD: To treat experimental CAG rats using mica monomer granule preparation with three different dosage-high, moderate and low level respectively. To observe the expression changes of mutant antioncogene-p53 gene-protein, oncogene p21, antioncogene p16 and anti-apoptosis gene bcl-2 in gastric mucosa of CAG rats by two-step ways of EnVision system in immunohistochemical method. RESULT: There was the tendency that mica monomer granule preparation with three different dosage could decrease the expression of p53 as well as p21, and mica had the obvious regulative effects on deletion of p16 and high-expression of bcl-2. It could also alleviate the inflammation of gastric mucosa and promote the regeneration of gland. CONCLUSION: The treatment and reversion action of mica on chronic atrophic gastritis is probably related with the regulative effect on the expression of gene associated with cancer. PMID- 16706024 TI - [Effects of slenderstyle Acanthopanax root-bark on cyclooxygenase]. AB - OBJECTIVE: To explore the effects of slenderstyle acanthopanax root-bark on cyclooxygenase in vivo and in vitro. METHOD: Contents of 6-keto-PGF1alpha in bovine aorta endothelial cells, PGE2 in mouse abdominal macrophages, and 6-keto PGF1alpha in rat stomach tissue were determined. The ulcer index in rat gastric mucosa was measured. RESULT: COX-1 and COX-2 were inhibited by slenderstyle acanthopanax root-bark, and the inhibitive rate of COX-2 was higher than that of COX-1 at the same concentration. Necrotic injuries in health gastric mucosa were not produced, but the injuries previously induced by the ethanol worsened. CONCLUSION: One of the antirheumatic mechanisms of slender-root-bark might probably be mediating through inhibition of cyclooxygenase. style acanthopanax PMID- 16706025 TI - [The expression of TLR4 in rat acute lung injury induced by lipopolysaccharide and the influence of Eucalyptus globulus oil]. AB - OBJECTIVE: To observe the distribution of toll-like receptor 4 (TLR4) in rats' respiratory tract. To study the influence of LPS and Eucalyptus globulus oil on the distribution of TMR4. METHOD: The Sprague-Dawley rats were intratracheally instilled with lipopolysaccharide (LPS,2 mg x kg(-1) per day) for two days to induce acute lung injury. The rats were sacrificed at 72 hours after LPS instillation. Lung morphology was studied. Leukocytes in Bronchoalveolar lavage fluid (BALF) were measured and TLR4 were detected by immunohistochemistry. RESULT: The result of immunohistochemistry showed that TLR4 distributed widely in common rats' respiratory tract. In the group of acute lung injury, the number of leucocyte in BALF was increased apparently, the inflammation in bronchus and bronchioles was more apparently than that of the control group in morphology. And the expression of TLR4 was reinforced in main bronchus and bronchioles. In the group of E. globules oil (300 mg x kg(-1)), the leucocyte number was decreased apparently in BALF, the inflammation was lightened and the expression of TLR4 decreased as compared with the group of models. CONCLUSION: The expression of TLR4 distributes widely in rats' respiratory tract. The stimulation of LPS can reinforce the expression of TLR4. The E. globules oil can reduce the increase of TLR4 induced by LPS in bronchioles. PMID- 16706026 TI - [Effects of total base of Sophora alopecuroides on expression of SOD, MDA, NO, MPO in rats with experimental colitis]. AB - OBJECTIVE: To investigate the effects of total base of Sophora alopecuroides on expression of SOD, MDA, NO, MPO in rats with experimental colitis (UC). METHOD: SD rats were randomized into 6 groups and colitis was induced by administrating 2,4,6-trinitrobenzesulphonic acid (TNB) recatlly in rats. Rats were given drug by stomach after the first day for 3 weeks, and sacrificed at 23rd day to determine the content of MDA and activities of SOD, NO, MPO in colonic mucosa, and to estimate the symptom and colonic histology. RESULT: In UC rats, SOD was significantly decreased whereas MDA, NO, MPO were increased when compared with the normal group (P < 0.05). In all treatment groups, Sophora alopecuroides could increase SOD, decrease MDA, NO, MPO (P < 0.05), and improve the symptoms and histological damages. CONCLUSION: The total dose of S. alopecuroides may ameliorates or alleviate the symptoms of UC through inhibiting oxygen free radical reaction, decreasing the generation of NO and exerting anti-inflammatory effects. PMID- 16706027 TI - [Effect of ray cartilage glycosaminoglycans (RCG) on the expression of MMP-9 in Lewis lung carcinoma]. AB - OBJECTIVE: To explore the effect of Ray cartilage glycosaminoglycans (RCG) on the expression of MMP-9 in Lewis lung carcinoma of mice. METHOD: The model of mice with Lewis lung carcinoma was induced. The experimental mice were randomly divided into normal saline group, RCG groups at varied concentrations and CTX group. Tumor growth state was observed, and tumor inhibitory rate of primary tumor and number of lung metastasis focus were measured. The expression of MMP-9 mRNA and protein in Lewis lung carcinoma was determined with RT-PCR and Western blot. RESULT: As compared with normal saline group, tumor growth curves in RCG groups were smooth, there were significant differences of inhibitory rates of primary tumor and number of lung metastasis focus between RCG groups and normal saline group, and MMP-9 mRNA and protein expression levels in RCG groups were reduced significantly. CONCLUSION: RCG can inhibit effectively the growth and metastasis of implanted Lewis lung carcinoma in C57BL/6 mice, which is probably attributed to reducing the expression of MMP-9 mRNA and protein. PMID- 16706028 TI - [Study on sini decoction in treatment of intestinal ischemia-reperfusion injury in rats: mechanism relating to oxygen radical and bcl-2 protein]. AB - OBJECTIVE: To investigate the protective effect of Sini decoction (SND) on intestinal mucosa in rats with intestinal ischemia reperfusion (I/R) and the mechanism relating to oxygen radical and Bcl-2 protein expression. METHOD: Thirty two SD rats of both sexes were randomly divided into 4 equal groups: (1) control group in which sham operation was performed; (2) model group in which intestinal I/R was produced by clamping super mesenteric artery(SMA) for 1 hour and declamping SMA for 3 hours; (3) SND low dose group in which SND(3 g x kg(-1) x d( 1)) was given via stomach tube for 3 days before operation; (4) SND high dose group in which SND(6 g x kg(-1) x d(1)) was given via stomach tube, for 3 days before operation. A strip of small intestine was taken from distal end of ileum for light microscopic examination, Chiu's score and the detection of intestinal water content (IWC). Apoptosis of intestinal mucosa cell was examined by TUNEL method. Superoxide dismutase (SOD) activity, malondisldehyde (MDA) concentration of intestinal mucosa were detected. The protein express of Bcl-2 of intestinal mucosa was analyzed by immunochemistory. RESULT: Intestinal /R resulted in histopathological changes in intestinal mucosa, increased Chiu's scores, apoptosis index, IWC and MDA content, and reduced SOD activity and the protein expression of Bcl-2 significantly (P < 0.01). The pretreatment of SND could attenuate the above changes significantly (P < 0.01), but there was no significant difference for the above variables between SND low dose group and SND high dose group (P > 0.05). Apoptosis index was significantly negatively correlative to SOD activity in model group and two SND groups. There were significantly negative correlation between apoptosis index and protein expression of Bcl-2 in model group and SND low dose group. CONCLUSION: SND can attenuate intestinal mucosa injury following intestinal U/R, which is related to reducing intestinal mucosa cell apoptosis by removing oxygen free radical and upregulating the protein expression of Bcl-2. PMID- 16706029 TI - [The influence of medicated serum with root of Crataegus cuneata on human sperm motility parameters in vitro]. AB - OBJECTIVE: To study the influence of root of medicated serum with the root of Crataegus cuneata on human sperm motility in vitro. METHOD: Sperm specimens of 16 asthenospermia patients were co-incubated with the medicated serum in vitro. The sperm motility characteristics were evaluated by computer-assisted semen analysis (CASA) at 5, 15, 30, 60 min and 120 min. RESULT: Compared with the control group, the medicated serum significantly increased the sperm progressive motility in 5 and 15 min (P < 0.05), and the motility and progressive motility were both increased significantly in 60 and 120 min (P < 0.01). The excitatory effect of the medicated serum on the sperm motility exhibited in a time-dependent manner. CONCLUSION: Root of C. cuneata medicated serum can improve sperm motility of asthenospermia patients in vitro. PMID- 16706030 TI - [Arnebia root oil promotes histological change and up-regulates bFGF and it's mRNA expression in the raw surface of the rabbits]. AB - OBJECTIVE: To explore the molecular biological mechanism of arnebia root oil in promoting wound surface healing by observing histological change and basic fibroblast growth factor(bFGF) mRNA expression in the wound surface tissues of 2 groups, as well as the wound surface healing rate. METHOD: Experimental model of incised-wound was produced on the back of 18 New Zealand albino rabbits. The wound surfaces were randomly divided into two groups, namely, experimental group and control group. The wound surfaces in the experimental group were treated by arnebia root oil and those in control group were treated by petrolatum gauze. Then raw surfaces were evaluated by the techniques of histology, histochemistry and electron microscope and the healing rates of the raw surfaces were compared between the two groups. Content of bFGF and it's mRNA expression in wound surface tissue was also measured by means of Western-blot and RT-PCR. RESULT: The wound surface healing rate in experimental group was higher than that in control group( P < 0.05). The fibroblast, collagen and blood capillaries were comparatively richer in experimental group as compared with those in control group, and similarly, the expression of bFGF mRNA was also significantly enhanced in the experimental group as compared with control group during the various periods of treatment. In addition, the changes in the expressions of bFGF and it's mRNA paralleled the changes of healing rates in the two groups. CONCLUSION: the present results showed that amebia root oil significantly can promote the healing of raw surfaces, which may be mediated by up-regulation of bFGF expression. PMID- 16706031 TI - [Distribution pattern and dynamics of Alnus nepalensis and Litsea glutinosa populations in Nomascus concolor habitat]. AB - Alnus nepalensis and Litsea glutinosa are the two common trees in the habitats of black-crested gibbons (Nomascus concolor) in Wuliang Mountain of Yunnan Province. With the nearest neighbor method and Heygi's competition index model for individual tree, this paper analyzed the distribution patterns of the two populations and their competition. The results showed that A. nepalensis had a random distribution at its different growth phases, while L. glutinosa, the companion species of A. nepalensis, exhibited the trend of random distribution clumped distribution-regular distribution. Interspecific competition and biological characters had significant effects on the distribution patterns of A. nepalensis and L. glutinosa. Regression age structure was observed in A. nepalensis population, while increasing age structure presented in L. glutinosa population. PMID- 16706032 TI - [Physioecological characteristics of walnut under different soil water conditions]. AB - The study with completely randomized design on Loess Plateau showed that soil water storage, water consumption, and water profit and loss had a positive correlation with the amount of irrigation. Managed with light, moderate and excessive pruning, soil water storage was increased by 0.2%, 0.5% and 0.9%, soil water consumption was reduced by 0.5%, 2.0% and 2.5%, while soil water profit and loss was increased by 150%, 190% and 210%, respectively. Under treatments with irrigation, covering with grass, mulching with plastic film, and pruning, soil water storage was increased by 4.4%, 1.2%, 1.6% and 0.5%, respectively. Irrigation, moderate pruning, covering with grass, and mulching with plastic film could increase leaf water potential and the angle between little leaf and stem, and excessive pruning could increase the chlorophyll content at most extent (0.27 mg x dm(-2)). The angle between little leaf and stem had a positive correlation with leaf water potential, which could be a reliable, direct and punctual index in evaluating the water condition of walnut and soil, and applied extensively in practice. PMID- 16706033 TI - [Critical tidal level for planting Kandelia candel seedlings in Xiamen]. AB - Plantable tidal flat is one of the most important factors affecting the survival rate of mangroves seedlings in forestation. In this paper, an experiment was conducted in the tidal zones of Umbrette Natural Reserve in the Dayu Island of Xiamen in May 2003 to investigate the critical tidal level for Kandelia candel forestation. The results showed that the tidal level of 0.99 m above the zero tidal level of the Huang Ocean was not suitable for planting K. candel seedlings, because the waterlogging time at this tidal level was longer than 8 h per-tide cycle, and the survival rate was lower than 50% . At 1.62 m above the zero tidal level of Huang Ocean, K. candel seedlings had the best growth and the highest photosynthetic assimilation, with a survival rate of 90%. At 1.31 m above the zero tidal level of Huang Ocean, K. candel seedlings could still grow well. It could be concluded that the tidal level of 1.62 m was optimal for planting K. candel seedlings, and the critical tidal level of K. candel seedlings in the coastal areas of Xiamen was not lower than 1.31 m above the zero tidal level of Huang Ocean, where the waterlogging time was not longer than 5.6 h per-tide cycle. PMID- 16706034 TI - [Dynamics of Sophora davidii population in a hilly region closed for thirty years in Loess Plateau]. AB - Taking wild grass ground as the reference, this paper studied the dynamics of Sophora davidii population on the sunny, semi-sunny and semi-shady slopes of hilly region in Loess Plateau after 30 years closed for reforestation, and analyzed the age structure, spatial distribution pattern, life table, and survival curve of the population. The results showed that S. dvidii population in the region belonged to progressive type, with more young and less old individuals. Among the three habits, semi-sunny slope had the highest population density. The survival curve of S. dvidii population was approached to Deevey 11 type. The population pattern was of clustered distribution, with the aggregating intensity increased with age. Water and light were the main environmental factors affecting the recovery and development of S. davidii community. The better recovery of S. davidii population suggested that the measures of closing the land for reforestation were efficient. PMID- 16706035 TI - [Water regulation characteristics and stress resistance of Caragana opulens population in different habitats of Inner Mongolia plateau]. AB - This paper studied the water regulation characteristics and stress resistance of Caragana opulens population in Helinger (semi-arid area) and Alashan (intensively arid area), aimed to understand the adaptation mechanisms of this population to arid environment. The results showed that compared with Helinger population, Alashan population had a higher content of osmotic adjustment substances in leaf cells, which resulted in a lower osmotic potential, and thus, stronger osmotic adjustment ability. The leaf water and free water contents of Alashan population were lower than those of Helinger population, while the bound water content and the ratio of bound water to free water of the former were higher than those of the latter. The leaf water potential of Alashan population was lower than that of Helinger population. Alashan population had a lower stomatal conductance than Helinger population, indicating that the former had stronger water adjustment ability. The malondialdehyde (MDA) content of Alashan population was higher than that of Helinger population, while the permeability of plasma membrane and the super-oxygen free radicals content of Alashan population were lower than those of Helinger population. It was suggested that Alashan population had a stronger ability in removing free radicals, and a higher stability of plasma membrane. C. opulens could adapt to arid environment through effective water adjustment and strong stress resistance. PMID- 16706036 TI - [Species diversity of bryophytes in West Tianmu Mountain of Zhejiang Province]. AB - In this paper, an investigation was made on the bryophytes at different altitudes of West Tianmu Mountain, with their species composition, similarity, and alpha- and beta-diversities. The results showed that at altitude 1100 m, the bryophytes under deciduous broad-leaved forest had the highest species number and richness, and the highest similarity with the bryophytes under deciduous broad-leaved shrub at 1300 m. The beta diversity index at altitude 800-1100 m was the largest, suggesting an obvious change and alternation of bryophyte species there. At altitude 1100 m, the species diversity of bryophytes was the highest, where should be the key area for bryophyte diversity conservation in West Tianmu Mountain. PMID- 16706037 TI - [Pollen assemblage and palaeo-vegetation of Late holocene fen in Dunhua of Jilin Province]. AB - Based on the study of pollen, stratum and 14C dating of Daqiao fen in Dunhua of Jilin Province, four pollen zones were distinguished, i.e., Pinus-Picea-Abies assemblage (2195 +/- 70 to appromimately 2045 +/- 70 B. P., Carex-Pinus BetulaCorylus-Juglans assemblage (2045 +/- 70 to appromimately 1745 +/- 70 B.P.), Pinus-Corylus-Carpinus-Carex-Ranunculus assemblage (1745 +/- 70 to appromimately 705 +/- 70 B.P.), and Pinus-Picea-Abies-Betula-Carex assemblage (705 +/- 70 to appromimately 1950 AD). The vegetation changed from coniferous forest (similar to the vegetation currently foundover 1100 m a.s. 1. in this area), through conifer broad-leaved mixed forest (similar to the vegetation currently found between 400 600 m a.s.l.) and conifer-broad-leaved mixed forest (similar to the vegetation currently found between 600-800 m a.s.l.), to conifer-broad-leaved mixed forest (similar to the vegetation currently found between 800-1100 m a.s.l.). Accordingly, Daqiao fen underwent the periods of gestation, fast development, expansion, and dieing out. PMID- 16706038 TI - [Inhibitory effect of pyrazole compound DL-1 on soil nitrification: a preliminary study]. AB - The study with soil incubation test under constant temperature and humidity showed that compared with DCD, DL-1 could significantly inhibit the oxidation of soil ammonium. In the first 3 weeks of incubation, DL-1 had an inhibitory effect of >70% on soil nitrification, with the best effect during 14-28 d of incubation. On the 14, 21, and 28 d of incubation, DL-1 with its dosage being 1.0 % of (NH4)2SO4-N decreased the soil NO3- -N content by 26.23%, 33.27% and 23.31%, respectively, compared with the same dosage of DCD, and by 71.12%, 69.10% and 55.14%, respectively, compared with no inhibitors applied. When the dosage of DL 1 accounted for 2.0% of applied N, soil nitrification was strongly inhibited, and soil NO3- -N content remained at a lower level till to the end (90 d) of incubation. PMID- 16706039 TI - [An on-the-spot sampling and survey method for soil nutrient cycling study]. AB - In this paper, an on-the-spot sampling and survey method for studying the soil nutrient cycling in a regional scale was discussed, with considering the principles of representation, reproducibility, randomness and timeliness. Firstly, some representative sampling areas in a definite region should be selected, based on the relevant hypsographic maps and airscapes. During the course of sampling soil and plant, a field survey related to the sampling sites should be carried out to understand the natural affecting factors on the soil nutrient cycling in a regional agro-ecosystem scale. Moreover, the farmers' basic status, crop production, and applied amount of fertilizers and their allotment should be also investigated. A case study of nutrient cycling in subtropical regions of China was carried out to approach the application of the method in the study of soil nutrient cycling in some regions. PMID- 16706040 TI - [Dynamics of soluble sugar and endogenous hormone contents in several steppe grass species during their germination period in spring]. AB - The study on the dynamics of soluble sugar and endogenous hormone contents in steppe grass species Elymus sibiricus, E. rectisetus, Leymus chinensis, L. racemosu and Stipa krylovii during their germination period in spring showed that high content GA and low content ABA played an important role for L. chinensis and L. racemosus germination, and the abundant soluble sugar storage in root made the rhizome grasses germinate earlier and grow stronger. The dynamics of ribosylzeatin was similar with that of soluble sugar in root, i.e., declined in early germination period, reached the lowest by the end of April, and ascended then, while the IAA and GA contents had a similar variation trend with the soluble sugar content in bud, i.e., all ascended gradually. There was a significant difference in the contents of GA, ABA and soluble sugar between rhizome grasses L. chinensis and L. racemosus and fibrous-root grasses E. sibiricus and E. rectisetus. PMID- 16706041 TI - [Establishment of snow disaster remote sensing monitoring and damage estimation systems in Altai pastoral region of Xinjiang]. AB - It is of significance to establish an integrated evaluation system of snow disaster in northern pastoral areas. Based on the NOAA satellite digital images, field observation data, and maps of grassland type and seasonal pastureland, this paper selected the winter and spring pasturelands in Aletai region of Xinjiang as the main area of snow disaster-remote sensing monitoring. With affecting factors of economy and the characteristics of natural resource distribution comprehensively analyzed, and using 3S techniques and field survey information, a fundamental information processing model for integrated evaluation of snow disaster was built up, and snow disaster-spatial evaluation indices and damage level systems were constructed. Natural and social systems and 20 indices were selected in snow disaster evaluation indicator system. Four principal factors, i.e., snow cover area, snow depth on grassland, persistence days of low temperature, and livestock death rate, were used as the grading indices of snow disaster damage level, and the models of snow disaster identification and loss estimatation were set up to quantitatively analyze snow disaster. The results indicated that the system could accurately reflect the details of snow hazard grade and the situation of a disaster in temporal and spatial scales, which would help to carry out the dynamic monitoring and scientific estimatation of big area's snow disaster in pastoral region. PMID- 16706042 TI - [Biomass structure and nitrogen and phosphorus contents of Calamagrostis angustifolia populations in different communities of Sanjiang Plain]. AB - Calamagrostis angustifolia is the dominant species in the typical meadow and marsh meadow communities of Sanjiang Plain. The study on its biomass structure and the N and P contents in its different organs showed that the biomass of different C. angustifolia organs in the two types of wetland communities was distinctly different, which could be described by Y = A + B1t + B2t2 + B3t3. The biomass of aboveground part and each organ presented single peak changing, with the maximum value of the latter occurred 15 d after. The F/C values were all less than 1, which was bigger in typical meadow than in marsh meadow. The total N and P contents in different organs of aboveground part all descended linearly in growth season, with the order of leaf > vagina> stem. The total N content in the roots of two C. angustifolia types was consistent, while that of total P was quite different. The contents of total N, NH4+ -N and NO3- -N, especially of NH4+ -N and NO3- -N, varied widely in different organs, with NH4+ -N/NO3- -N >1. Root was the important storage of N and P, but the storage of N and P in stem, leaf and vagina fluctuated greatly. The N/P ratios of two C. angustifolia types were all less than 14, which implied that N might be the limiting nutrient of C. angustifolia, and the limitation degree was higher in typical meadow than in marsh meadow. PMID- 16706043 TI - [Allelopathy of invasive weeds: a simulation study with cellular automata model]. AB - Cellular automata model is a simulation approach to describe the complicate behavior of a system, and suitable to study the spatial and temporal dynamics of plant community. In this paper, the model was used to simulate the different sensitivity toall invasion process of an allelochemicals-containing exotic species to the community of two native species with different sensitivity to allelochemicals, and the spatial and temporal dynamics of native and invasive species. The simulation was conducted by biological response and negative exponential distribution models, and the results showed that exotic species could successfully invade the community of two native species with different sensitivity to allelochemicals, but only coexist with one sensitive and one resistant species. The resistance of plant community to invasive weeds depended on its species function structure. PMID- 16706044 TI - [Emergy analysis of mulberry-silkworm ecosystem in China]. AB - Mulberry-silkworm ecosystem is one of the important agro-ecosystems in China. Based on the principles and methods of emergy analysis, this paper studied the interior structure of mulberry-silkworm ecosystem and its relationship with exterior environment and economy. Some emergy indices for this ecosystem were quantitatively calculated, and compared with those of the agro-ecosystem in China. The results showed that the emergy investment ratio, emergy yield ratio, environmental loading ratio and emergy sustainability index was 3.78, 4.68, 0.18 and 26.0, respectively, suggesting the low environmental pressure and good ecological benefit in mulberry-silkworm ecosystem in China. Hi-technology was required to further decrease the labor force input and enhance the comprehensive utilization of sericultural resources. PMID- 16706045 TI - [Emergy value evaluation on rice-duck organic farming mode]. AB - Employing emergy value evaluation method, this paper compared the ecological and economic benefits of rice-duck organic farming system (mode I) and conventional rice-wheat rotation system (mode II) in Shanghai suburb. The results showed that mode I had better emergy benefits, higher self-organizing ability and sustainability, and higher product safety. The net emergy yield ratio (EYR), feedback ratio of yield emergy (FYE), and emergy sustainable index (ESI) of mode I were respectively 1.57, 14.1, and 8.71 times as much as those of mode II, and the emergy index of product safety (EIPS) was 0 in mode I but -0.66 in mode II. The emergy investment ratio (EIR) and environmental loading ratio (ELR) of mode I were 40.1% and 18.3% of mode II, respectively, suggesting that mode I had less environmental pressure than mode II. Mode I had lower economic benefits than mode II. The economic output, gross income, and net income of mode I were 15.7%, 9.6%, and 29.6% less than those of mode II , respectively. As for Em dollars, the output, gross income, and net income of mode I were respectively 50%, 102.6%, and 136.4% higher than those of mode II. With the system optimization and the development of organic food market, mode I had the potential to improve its economic benefits. PMID- 16706046 TI - [Effects of straw mulching and irrigation on solar energy utilization efficiency of winter wheat farmland]. AB - The study showed that straw mulching decreased the basic seedlings and tillers of winter wheat and the leaf area index (LAI) at earlier growth stage, but increased the LAI at latter growth stage. Straw mulching and irrigation reduced the transmittance and reflectance of PAR, resulting in the increase of PAR capture ratio mainly at the height of 40-60 cm. The solar energy utilization ratio of grain was decreased by straw mulching, while that of stem and leaf was increased. The total solar energy utilization efficiency of winter wheat could also be increased by straw mulching. PMID- 16706047 TI - [Effects of nitrification inhibitors on nitrate content in soil and pakchoi and on pakchoi yield]. AB - A field experiment was conducted on two soil types in the Changsha suburb of Hunan Province to study the effects of hydroquinone (HQ), dicyandiamide (DCD) and thiourea (TU) on the nitrate content in soil and pakchoi and on the yield of pakchoi. The results showed that all the test nitrification inhibitors could significantly decrease the nitrate content in soil and pakchoi during whole growth stage, among which, DCD had the best effect, but the effect was differed on different soil types. Nitrification inhibitors could increase pakchoi yield, DCD was also the best one, and the effect was significantly better on vegetable garden red soil than on vegetable garden alluvial soil. The nitrate content in soil and pakchoi was the highest about 40 days after pakchoi transplanting. PMID- 16706048 TI - [Effects of potassium application rate and its supplemental proportion on dry matter accumulation and potassium absorption of flue-cured tobacco]. AB - A field trial with flue-cured tobacco conducted on a red paddy soil showed that the dry matter accumulation in the tobacco reached a peak in 50-60 days after transplanting, but was lower within 30 days or over 80 days after transplanting. The dry matter accumulation was increased with increasing K application rate and its supplemental proportion. The K content in the tobacco got its peak in 30-50 days after transplanting, and then went down, with the K content at maturing stage being as half as that in the peak period. The K content of the tobacco in each growth stage was obviously higher with 315 kg x hm(-2) K application than with 225 kg x hm(-2) K application. At maturing stage, the dry matter and K contents in the tobacco decreased with 225 kg x hm(-2) K application, but increased with 315 kg x hm(-2) K application, and the K content was higher with 70% K supplemental proportion than with 50% and 30% K supplemental proportion. PMID- 16706049 TI - [Effects of bagging on pigment, sugar and endogenous hormone contents of Cara Cara orange flesh]. AB - This paper studied the effects of bagging during the period from young fruit formation to fruit coloration on the contents of pigment, sugar and endogenous hormone in Cara Cara orange flesh. The results showed that bagging could significantly increase the lycopene and beta-carotene contents of matured fruit, but didn't have any effects on its GA and ABA contents. The GA content decreased rapidly during fruit-expanding period, and maintained at a lower level in the period from fruit coloration to maturing. The ABA content reached the maximum when the bag was removed, decreased rapidly then, and there was a small peak before fruit maturation. Glucose and fructose contents were decreased, while sucrose content was increased significantly. No significant change was observed in total sugar content. PMID- 16706050 TI - [Behaviors of Propylaea japonica (Coleoptera: Coccinellidae) in searching and preying cotton aphid Aphis gossypii (Coleoptera: Coccinellidae) in the cotton plant]. AB - The study showed that the searching activity of P. japonica on cotton aphid Aphis gossypii decreased gradually with decreasing aphid density. When enough A. gossypii existed, P. japonica had a preferential searching and preying activity on the top of cotton plant. There was no significant difference in the preying activity of P. japonica at different positions of cotton plant, while the searching activity of the ladybird beetle was significantly lower on cotton M9101 than on cotton Zhongmian 13. The preying behavior of P. japonica was significantly correlated with the density and distribution of A. gossypii. PMID- 16706051 TI - [Community diversity of butterfly in Houhe National Nature Reserve]. AB - In 2003-2004, a field investigation was made on the composition of butterfly community in the Houhe National Nature Reserve of Hubei Province, and the diversity, evenness and dominance of the community on the levels of family, genus and species in spring, summer and autumn were studied by the method of alpha diversity. The results showed that the species richness, diversity, and dominance indices were the highest in summer, while the evenness index wasthe highest in spring. Nymphalidae was the highest in richness at the level of families, with the value of 32. PMID- 16706052 TI - [Avian diversity in Wuhu City in summer and winter]. AB - From May 2004 to February 2005, an investigation was made on the avian communities in four typical habitats, i.e., village and farmland, urban woodlot, floodplain, and riverine wetland in Wuhu City, with the related diversity indices analyzed. A total of 117 species belonging to 15 orders and 31 families was recorded. The avian species and G-F index in village and farmland, urban woodlot, and floodplain were higher in summer than in winter, while the species number, Shannon-Wiener index and Pielou index in riverine wetland were significantly higher in winter than in summer. The avian species and G-F index in urban woodlot were the highest, and the Shannon-Wiener index and Pielou index in floodplain were higher than those in other three habitats. The highest density was observed in village and farmland in summer and in urban woodlot in winter, being 31.1140 and 42.9160 ind x hm(-2), respectively. The avian species, density, Shannon Wiener index and G-F index in riverine wetland were the lowest. Avian diversity could be enhanced by increasing spatial heterogeneity and reducing human disturbance. PMID- 16706053 TI - [Modeling of Spodoptera litura nuclear polyhedral virus epidemic at different temperatures]. AB - The study on the nuclear polyhedral virus epidemic of Spodoptera litura under effects of temperature showed that at above 29 degrees C, S. litura larvae started to die 2-3 d after virus-inoculation, and reached the peak 4-5 d after the inoculation. The mortality and death velocity of virus-infected host larvae were increased with increasing incubation temperature. No thermo-inhibition temperature for the virus was observed in the test temperature range, while there existed a constant temperature range, i.e., 29-35 degrees C, for the highest mortality of infected host larvae. Complementary log-log regression model could well simulate the daily distribution of host larvae mortality, and the cumulative disease death time could be described by Gompertz model. The revised biophysical model could well simulate the relationship between S. litura larvae mortality and temperature, and be applied to virus epidemic forecasting. PMID- 16706054 TI - [Predation of Typhlodromus bambusae Ehara on Sehizotetranychus bambsae]. AB - The simulation study under laboratory condition showed that the functional responses of Typhlodromus bambuse to Schizotetranychus bambuse at different development stages could be expressed by the curve of Holling Type II. The number of prey consumed by predator quickly increased with initial prey density, but soon leveled off. At 25 +/- 0.5 degrees C, the handling time (Th) of T. bambuse to S. bambusae at the stages of egg, larva, protonymph, deurtonymph and female was in order of egg02C-MB >Xinchun 6 >Bafeng 1> Yongliang 4> Neimai 19> Menghua 1 using the intrinsic rate of increase (r(m)) as an index of wheat antibiosis to aphids, which was correspondent well with that in fields. PMID- 16706071 TI - [Hepatic encephalopathy and nodular liver in an adolescent]. PMID- 16706072 TI - Developments in Barrett's esophagus. PMID- 16706073 TI - Clinical characteristics and long-term outcome of patients with refractory sprue diagnosed at a single institution. AB - BACKGROUND: Refractory sprue (RS) is a rare and severe celiac-like enteropathy not responding to a strict gluten-free diet. Although prognosis is generally poor, little is known about the long-term outcome of patients. AIM: to report baseline characteristics and long-term outcome of a series of patients diagnosed and treated in a single institution. MATERIALS: We report a retrospective cohort of 25 consecutive patients (15 females; mean age 46 yr; range 28-71) diagnosed with RS based on the presence of a non-responsive celiac-like enteropathy. All patients were intensively treated with a gluten-free diet, steroids, nutritional support and immunosupression. RESULTS: Clinical and biological characteristics of patients suggest that, at least, 24 patients had clear evidences of celiac disease. HLA DQ2/DQ8 genes were present in all the 24 patients typed and autoimmune enteropathy was excluded in all. According to the genotyping, 12 patients had a polyclonal lymphocyte population (RS type I) and 13 exhibited monoclonal TCR-gamma gene rearrangements (RS type II). Sixteen patients had evidence of ulcerative jejunitis (UJ) (7 in RS type I and 9 in type II). Overall median follow-up time after diagnosis of RS was 29 mo/patient (range 7 to 204) (45 mo for type I and 24 mo for type II). Overall mortality was 48% (12 patients), 6 in each type. Eight patients with UJ (50%), 3 with lymphoma (two T cell and one B-cell type) and 4 (44%) without ulcers died during follow-up. The causes of death were sepsis in the context of a progressive deterioration but without overt malignancies (n=5), vascular causes (n=3) and severe malnutrition (n=1). Three- and 5-yr survival rate after diagnosis of RS for the overall population was 60% and 56%. There was no differences between type I (67%, 58%) and type II RS patients (54% for both periods). Patients with UJ had lower but non-significant 3- and 5-yr survival rates (56% and 50%, respectively) compared with patients without ulcers (78% and 66%). Survivors had a favorable outcome. While 11 patients persists asymptomatic, two other cases still have mild diarrhea and one low body weight. CONCLUSIONS: We confirm that RS is a severe celiac disease-related disorder with very high mortality. Diagnosis of overt lymphoma (12%) in our long-term follow-up was not as frequent as was reported by other groups. A proportion of patients persist in good health for a long time irrespective of the nature of the IEL infiltration or the presence of UJ. PMID- 16706074 TI - Multidrug resistance gene (MDR-1) expression in the colonic mucosa of patients with refractory ulcerative colitis. AB - BACKGROUND: P-Glycoprotein (P-gp), a product of the MDR-1 gene, is a transmembrane efflux pump involved in drug transport, first described in cancer refractoriness. In the normal bowel P-gp is detectable on superficial epithelial cells, but has not been described in crypt epithelium. The role of P-gp and its intestinal expression in steroid-refractory ulcerative colitis (UC) are controversial. AIM: to compare P-gp immunostaining pattern in colonic epithelial cells of steroid-refractory versus steroid-responder UC patients. METHODS: P-gp was assessed by immunohistochemistry in rectal biopsies obtained from 19 patients with active UC, including pre-surgical samples from 11 refractory patients who underwent colectomy, and 8 responders. We devised a 5-point (0-4) score, according to the percentage of epithelial surface with positive immunostaining in the superficial and crypt epithelium (apical, lateral and cytoplasmic areas). RESULTS: Compared with responders, steroid-refractory patients had significantly higher immunostaining scores in the superficial epithelium, both in apical (2.8+/ 0.5 versus 1.1+/-0.5, p=0.023) and cytoplasmic cellular areas (2.7+/-0.5 versus 1.2+/-0.5, p=0.032). Positive immunostaining of the superficial epithelium was frequently detected in refractory patients (apical: 9/11 cases, cytoplasmic: 10/11 cases) but was only observed in 4/8 responders. P-gp was also detected in similar areas of the crypt epithelium in 6/11 refractory patients, while it was infrequent in the group of 8 responders (1 apical 1 case, cytoplasmic 2 cases). Samples from the mucosa of normal ileal pouch-anal anastomoses obtained several years after the surgical procedure had a P-gp immunostaining pattern which was similar to that of rectal samples from patients with refractory UC. CONCLUSIONS: These results suggest a critical role of P-gp overexpression in steroid refractory UC. PMID- 16706075 TI - [Gastric emptying of liquids in rats submitted to acute myocardial infarction]. AB - Gastric Emptying (GE) is food transition from the stomach to the upper small intestine. Haemodynamics changes modify gastric emptying and the gastroduodenal motor activity. Myocardial Infarction is a pathological condition in which there are haemodynamics changes. Therefore, the objective of the present investigation was to study the effect of acute myocardial infarction on rat gastric emptying of an isosmotic and inert liquid meal. We conducted a study involving male Wistar rats (varying in body weight from 220 g to 250 g), that were allowed to habituate in a laboratory environment and then divided into 3 groups: group INF, rats in which the myocardial infarction was produced by left anterior coronary artery ligation; group SH, rats in which the myocardial infarction was simulated without ligating the anterior coronary artery; and group NA, rats which were not given any anesthesia or surgical procedures. After group constitution, animals were fasted with access to water ad libitum. 24 hours later, the Gastric Retention (GR) of 1.5 ml/100 g animal weight of a test meal of 0.9% NaCl plus the marker phenol red was evaluated. GE was indirectly evaluated in conscious animals, through determining the percentage of gastric retention (% GR) of a test meal, measured 10 minutes after orogastric infusion. Results of the present study showed that animals from the INF group presented GR (median=48.0%), significantly higher than the NA group (30.6%) and SH group (36.6%). No statistical difference in GR was observed between the SH and NA groups. Infarcted area, expressed in percentages, in animals of the INF group was 51.3+/-4.7% (mean SEM, N=17) which not presented correlation with results of GR of the same group (r=-0.05). From a caloric viewpoint, the results suggested that acute myocardial infarct in rats induce delay of the gastric emptying of an isosmotic and inert liquid meal. PMID- 16706076 TI - [Development of porphyria cutanea tarda in a chronic hepatitis C patient with indetectable viremia under treatment with peg-interferon plus ribavirin]. AB - Porphyria cutanea tarda (PCT) is considered an extra-hepatic manifestation of HCV infection. The frequency of this association varies according to different authors and the mechanism by which the virus can trigger this disease is not yet clear. We present a 47-year-old-man with chronic hepatitis C genotype 1b who, during the treatment with peg-interferon alfa 2b plus ribavirina, with no detectable viremia at weeks 12th, 24th, and 48th, developed dermatological photosensitive lesions at week 44th. With a presumptive diagnosis of PCT a cutaneous/skin biopsy was performed as well as a porphyrin dosage with urine porphyirins of 4185 microg/24 hs (nv<250). The chromatographic analysis revealed the typical PCT pattern thus confirming the diagnosis. The hemochromatosis HFE gen evaluation showed heterozigotus character mutations (H63D and C282Y) a frequent association in patients with iron overload and PCT. The antiviral treatment of the HCV infection can improve the clinical-humoral manifestations of PCT. The novo occurrence of PCT was recently reported during chronic hepatitis C treatment with interferon and ribavirin, but no cases of late appearance of PCT in patients with no detectable viremia were reported. The mutation of the gen HFE in our patient and the hemolysis caused by ribavirin can be related to the development of the disease, but the iron overload because of ribavirin use is also controversial. This is another example of the complexity of this association. PMID- 16706077 TI - [GERD related respiratory symptoms: diagnosis and treatment]. AB - Gastroesophageal reflux disease can cause respiratory symptoms. These symptoms are triggered by reflux events that reach the pharynx, causing microaspiration or through vagal reflex. Respiratory symptoms can be vague and coexist with gastroesophageal reflux disease, without a real link between the two entities. To effectively treat these patients, it is important tofind an association between the two diseases. Work up should include the diagnosis of reflux disease, the diagnosis of pharyngeal reflux events--microaspiration--and, if possible, of laryngeal injury. Once the diagnosis has been established, an effective therapy must be offered to the patient. In these patients, medical treatment is less effective when compared to the results in the population with typical symptoms. This may be due to the fact that non-acid reflux episodes are causing the respiratory symptoms or as a result of an irreversible damage generated in the airway. Antireflux surgery is an effective therapy that reduces both acid and non acid reflux events. This article describes the different diagnostic tests as well as the results obtained with surgical treatment in this population. Additionally, it describes potential applications of esophageal and pharyngeal impedance monitoring in these patients. PMID- 16706078 TI - Crisis! PMID- 16706079 TI - Am I my brother's keeper? PMID- 16706080 TI - More thoughts on medical missions. PMID- 16706081 TI - Hurricane Katrina response in Knoxville: lessons learned. PMID- 16706082 TI - Ask TMA. How can I reduce the hassle of legal depositions? PMID- 16706083 TI - TN doctors appointed to Medicare Health Support Medical Advisory Board. PMID- 16706084 TI - Mind medicine: treating mental illness in Tennessee. PMID- 16706085 TI - Consequences of poor evaluation. PMID- 16706086 TI - Here to help: annual PHP activity report. PMID- 16706087 TI - Succession planning vital for physician practices. PMID- 16706088 TI - Strategies for managed care negotiating. PMID- 16706089 TI - Perceived impact of TennCare reform on patients' health in a medical teaching practice. AB - From 1994-2005, TennCare, Tennessee's innovative Medicaid managed care program, dramatically expanded coverage to around 1.3 million Tennesseans (roughly 23 percent of the state population) by employing cost savings achieved through capitation and managed care. Rising healthcare costs and a static state budget resulted in program downsizing that started in mid-2005. This cross-sectional survey sought to document the perspectives of TennCare enrollees before disenrollment about the likely impacts of changes in TennCare coverage and benefits. In February 2005, a convenience sample of 89 patients served by an internal medicine resident staffed teaching practice in Memphis, TN, participated in a survey to assess their perspectives about the potential impact of the upcoming disenrollment and benefits limitations. Ninety percent or more expressed concerns that loss of TennCare would lead to health problems, difficulty with paying for prescriptions and difficulty finding alternative health insurance. This survey suggests that before disenrollment, most people served by TennCare believed that loss of TennCare would have serious negative consequences on their health. Further studies are needed to assess the true impact of the disenrollment and benefit cuts and the effectiveness of alternative safety net services for vulnerable Tennessee citizens. PMID- 16706090 TI - The uninsured in Tennessee. PMID- 16706091 TI - The use of restraints in psychiatric settings. PMID- 16706092 TI - Standards of identity: using pizza logic to re-brand TMAA. PMID- 16706093 TI - Professor Panayotis I. Chrysospathis, MD, PhD, FACS, FICS: the golden sword and the great ambassador of Greek surgery. PMID- 16706094 TI - Hypovolemic shock caused by a true aneurysm of the pancreatico-duodenal artery: case report and review of the literature. AB - True pancreatico-duodenal artery aneurysm (PDAA) is a rare condition that since 1973 has been described in only 54 cases. It is frequently associated with celiac axis stenosis and often present with rupture. Even if most PDAAs that are not ruptured are asymptomatic and are diagnosed during investigation for other diseases, they may have some symptoms, such as chronic abdominal discomfort and an abdominal pulsating mass, that can be helpful for diagnosis. The treatment of this condition has evolved in time from a merely surgical one to an angiographic noninvasive approach. We present a case of a PDAA that manifested with sudden hypovolemic shock requiring an emergency operation, and through a review of the literature, we discuss the different diagnostic/therapeutic protocols to use in different situations. PMID- 16706095 TI - Ureteral stents and their current use. AB - Stents in urology have been used for a long time in the management of nephro ureterolithiasis. However, the indications for stent insertion have increased during the last few years and they are now used both diagnostically and therapeutically in a variety of cases. A brief history of the name is presented along with a review of the indications and the complications of stent usage. PMID- 16706096 TI - Follow-up and recurrence of T1 colorectal cancer. AB - This study was performed to clarify the clinical and pathological characteristics of T1 colorectal cancer. From 1987 to 2003, a total of 223 patients with T1 colorectal cancer were treated at our institute. Disease recurrence occurred in eight (3%) of these cases. The original sites of cancer were the ascending colon (n = 1), sigmoid colon (n = 2), and rectum (n = 5). We performed three local resections per anus and five bowel resections with lymph node dissection. In cases showing recurrence after resection, lymph node metastasis was observed more frequently (four of five cases; 80%) compared with the cases showing no recurrence (11%). The average recurrence time was <36 months in seven of the eight cases (88%). Local recurrence occurred in five of the eight cases (63%). We conclude that careful follow-up is necessary in cases of T1 colorectal cancer in the first 3 years after treatment. PMID- 16706097 TI - Colovesical fistula complicating diverticular disease: one-stage resection. AB - Colonic diverticular disease is common in developed countries, and its prevalence increases with age. Most affected individuals remain asymptomatic throughout their lives, and relatively few patients require surgical intervention for obstructive or inflammatory complications. Colovesical fistula is the most common type (65%) of fistula associated with colonic diverticular disease. Primary resection of sigmoid colon with colorectal anastomosis performed as a one-stage procedure is its definitive treatment and can be performed safely--as simple closure, using an omental flap, or through resection and closure of bladder defect--in 90% of the patients. We report our experience with four patients suffering from colovesical fistula who were treated with primary resection of sigmoid colon and colorectal anastomosis performed as a one-step procedure. In our experience, diverting colostomy or Hartmann intervention is not recommended because of the lack of fistula definitive resolution and the possibility of additional complications. PMID- 16706098 TI - Pancreaticoduodenectomy for common bile duct cancer in a patient with situs inversus totalis: a case report. AB - We describe the first case of successfully performed pancreaticoduodenectomy to treat common bile duct cancer in a patient with situs inversus totalis. A 65-year old man was admitted to our hospital with the working diagnosis of obstructive jaundice. He had a history of cholecystectomy, and situs inversus totalis had been diagnosed at that time in another hospital. Preoperative work-ups with ultrasonography, computed tomography, and percutaneous cholangiography were followed by drainage lead to the diagnosis of common hepatic artery arising from the superior mesenteric artery. He underwent curative pancreaticoduodenectomy without complications. The postoperative course was uneventful, and he was discharged on postoperative day 29. He is doing well, with no evidence of recurrence 40 months after surgery. Deliberately performed preoperative imaging studies, especially selective angiography, were essential for the surgical resection of bile duct cancer in this patient with situs inversus totalis. PMID- 16706099 TI - Pulmonary carcinosarcoma: report of a case with review of the literature. AB - Pulmonary carcinosarcoma is a very rare tumor, and its prognosis is usually worse than pulmonary carcinoma. We present a case with gigantic carcinosarcoma (10 x 7.7 x 7.2 cm) located in the right upper lobe, in which the discordance between the brief clinical course with limited symptoms and its large size are remarkable. No definite diagnosis was made preoperatively. He underwent right upper lobectomy and mediastinal lymph node dissection through thoracotomy. The postoperative course was smooth, and no tumor recurrence or metastasis was noted during the 9-month follow-up period. In the literature, carcinosarcoma have been divided into the endobronchial and peripheral types, which have different histological pictures. The controversy about pathology, diagnosis, and treatment of carcinosarcoma is also discussed. PMID- 16706100 TI - Retroperitoneal para-aortic paraganglioma. AB - Paragangliomas or chemodectomas are neoplasms that arise from neural crest cells and histologically resemble their adrenal counterpart, the pheochromocytoma. Seventy-one percent of the extra-adrenal paragangliomas are located in the superior or inferior para-aortic area. This tumor usually presents as an abdominal mass producing back pain. Tumor localization has improved remarkably through the use of computed tomography and I131MIBG (metaiodobenzylguanidine) scintigraphy, particularly when tumors are hormonally active. Surgical excision remains the mainstay of treatment, although vascularity can at times make excision difficult. PMID- 16706101 TI - Surgical complications caused by inaccurate scotopic preoperative pupillary measurement in cataract and lasik surgery: case report and review of the literature. AB - Accurate scotopic preoperative pupillary measurements in both cataract and lasik surgery cases are critical to avoid in individuals with naturally briskly dilating pupils under scotopic conditions postoperative visual disability related to low light reflections, disabling illuminance, glare disability, halos, starbursts, and diplopia with night vision. Both edge reflections of the pseudophakos implant in cataract surgery and the ablated corneal margin in lasik surgery can contribute to serious visual disability in patients who have larger scotopic pupils exposing the edge of the implanted lens or the surgical optical zone of the ablated cornea to the entrance pupil. A case is presented in which cataract surgery was performed initially with a 5.5-mm-diameter optic lens in one eye and later a 7.0-mm-diameter optic lens in the other eye. Because of a naturally briskly dilating pupil of 7.0 mm, the eye with the smaller implant experienced significant night vision difficulties because of edge reflections of the exposed implant. The other eye in the very same patient with the larger lens implant was asymptomatic. This case underscores the importance of accurate preoperative pupillary measurements under scotopic conditions to avoid this preventable disabling surgical complication in both cataract and lasik cases. PMID- 16706102 TI - Firearm-related injuries: clinical considerations on 1326 cases. AB - We evaluated the characteristics of 1326 patients with gunshot wounds, between 1 January 2001 and 31 December 2002, considering sex, ages, circumstances, involved regions, hospitalization, surgical procedures performed, complications, sequelae, and mortality. They were analyzed by a retrospective statistical study and compared applying the Z test. The results showed that only 7% were women; mean age was 28.35 years, and there were significant differences in self-inflicted injuries depending on age and sex. Wounds were usually located in one body region. Most frequent surgical procedures were pleural drainages and "surgical toilette" in open fractures. Infections and neurological sequelae were the most frequent complications. The mortality rate was 6.18%. More people died at the scene than at the hospital. Increasing violence, all over the world, brings a higher number of people with gunshot wounds to hospitals. Sequelae cause a complete change in people's lives, not only through the resultant disability, but also by causing work and social problems. PMID- 16706103 TI - Role of video-assisted thoracic surgery for the treatment of myasthenia gravis: extended thymectomy by median sternotomy versus the thoracoscopic approach with sternal lifting. AB - The role of video-assisted thoracic surgery (VATS) thymectomy is still being studied, and many surgeons remain skeptical of the value of this recent option. We made a retrospective evaluation to ascertain whether VATS-extended thymectomy is as reliable as standard median sternotomy in the treatment of myasthenia gravis (MG) and whether the endoscopic procedure presents any advantages for patients. Eighteen consecutive patients requiring extended thymectomy for MG were treated between April 1997 and September 2003 at our hospital. Nine patients received VATS-extended thymectomy, and the remaining nine patients received standard extended thymectomy by sternotomy. In the VATS group, the anterior mediastinal space was well visualized by sternal lifting. The mean operative time was 268.3 +/- 51.1 minutes in the VATS group and 177.3 +/- 92.5 minutes in the sternotomy group. Operative time was significant longer in the VATS group than in the sternotomy group (P < 0.05). The mean operative bleeding was 68.6 +/- 47.8 ml in the VATS group and 154.1 +/- 109.0 ml in the sternotomy group. Operative bleeding was significantly less in the VATS group than in the sternotomy group (P < 0.05). There was no significant difference between the two groups with regard to postoperative duration of chest tube or the level of serum C-reactive protein on the first operative day. There was a downward trend in nicotinic acetylcholine receptors antibody levels after thymectomy compared with before thymectomy in both groups. VATS thymectomy should be considered a valid alternative to the established approaches aimed at achieving a "curative thymectomy" in patients with MG. PMID- 16706104 TI - Staged approach to the urachal cyst with infected omphalitis. AB - Embryonal urachus exists as a cord-like structure between the urinary bladder and the umbilicus. In some cases of urachal cysts at the level of the navel, no special symptoms are detected during childhood, but spontaneous drainage at the navel may occur after adolescence, which is called an infected urachal cyst. Especially in cases accompanied by infected omphalitis, no constant opinion has been established to choose either initially curative resection or staged incision. In this study, we evaluated the characteristics of patients with urachal cysts who underwent the staged approach. Twenty patients (14 men and 6 women) with urachal cysts complicated by infected omphalitis were treated in our hospital. Staged surgery was performed for 18 patients. Neither recurrent omphalitis nor subsequent urachal carcinoma was observed. It is desirable that urachal cyst accompanied by intractable omphalitis should be treated by conservative therapy, conducted image diagnosis, and chosen staged surgery. PMID- 16706105 TI - Unusual presentation of an appendiceal malignancy. AB - Primary malignant epithelial tumors of the appendix are uncommon. The most common presentation of appendiceal malignancy is right lower abdominal pain suggestive of acute appendicitis. Presentation caused by loco-regional spread with involvement of neighboring organs is rare. We present the case of a 48-year-old woman with an appendiceal malignancy who presented with symptoms and signs suggestive of complicated diverticular disease with an enterovaginal fistula. From a review of the literature, this is the first report of an appendiceal malignancy presenting in this manner. PMID- 16706106 TI - Effects of male sex hormones on gender identity, sexual behavior, and cognitive function. AB - Androgens, the male sex hormones, play an essential role in male sexual differentiation and development. However, the influence of these sex hormones extends beyond their roles in sexual differentiation and development. In many animal species, sex hormones have been shown to be essential for sexual differentiation of the brain during development and for maintaining sexually dimorphic behavior throughout life. The principals of sex determination in humans have been demonstrated to be similar to other mammals. However, the hormonal influence on sexual dimorphic differences in the nervous system in humans, sex differences in behaviors, and its correlations with those of other mammals is still an emerging field. In this review, the roles of androgens in gender and cognitive function are discussed with the emphasis on subjects with androgen action defects including complete androgen insensitivity due to androgen receptor mutations and 5alpha-reductase-2 deficiency syndromes due to 5alpha-reductase-2 gene mutations. The issue of the complex interaction of nature versus nurture is addressed. PMID- 16706107 TI - [HSF1 inhibits heat stress-induced apoptosis in Raw264.7 macrophages]. AB - OBJECTIVE: To observe the effect of heat shock factor 1 (HSF1) on heat stress induced apoptosis in Raw264.7 macrophages. METHODS: Raw264.7 cells transfected with pcDNA3.1 and pcDNA3.1-HSF1 were exposed to heat stress (42.5 degrees C +/- 0.5 degrees C) for 1 h and recovered at 37 degrees C for 6, 9, 12, and 24 h respectively. Flow cytometry (FCM), Hoechst 33258 staining and DNA ladder assays were performed to assess the apoptosis. RESULTS: After heat stress, FCM showed that apoptotic cells were increased significantly and reached the peak at 9 h in Raw 264.7 cells transfected with pcDNA3.1, and were characterized with classical morphologic changes including apoptotic body and nuclear condensation. Agarose gel electrophoresis showed that "DNA ladder" could be observed clearly at 6, 9, and 12 h after the heat stress. But the overexpression of HSF1 could reduce the number of apoptotic cells and inhibit DNA fragmentation. CONCLUSION: HSF1 can inhibit heat stress-induced apoptosis in Raw264.7 macrophages. PMID- 16706108 TI - [Screen of inflammatory genes regulated by heat shock factor 1 and corroboration with SOCS3 gene]. AB - OBJECTIVE: To screen the inflammatory mediators genes regulated by HSF1, and explore the mechanism of downstream genes regulated by HSF1. METHODS: HSF-/- and HSF1+/+ mice were injected with 15 mg/kg LPS intraperitoneally (ip), respectively, and were treated as previous after HSR. The total RNA of lung tissues were extracted and filtrated by SuperArray gene Microarry. The promoter of candidate genes were analyzed by transcription element search software to search for heat shock element (HSE). Select the suppressor of cytokine signaling 3 (SOCS3) with HSE. Macrophage cells were stimulated with 400 ng/mL LPS, and were treated as previous after HSR, then the total RNA was extracted respectively. RT PCR and northern blot assay were performed to detect the expression levels of SOCS3 mRNA. RESULTS: Fifteen genes were repressed by HSF1, including 9 genes with complete HSE. Eleven genes were accelerated by HSF1 possibly, including 8 genes with complete HSE. The promoter of SOCS3 gene contained one complete HSE. LPS stimulation obviously increased the levels of SOCS3 mRNA in macrophages of RAW264.7 mice, which was inhibited by HSR and over-expression of HSF1. CONCLUSION: HSR or HSF1 inhibits LPS induced expression of SOCS3 mRNA; HSF1 might inhibit LPS-induced expression of SOCS3 mRNA by binding to HSE in the promoter of SOCS3 gene. PMID- 16706109 TI - [Immortalization of embryonic fibroblasts in heat shock transcription factor 1 knockout mouse]. AB - OBJECTIVE: To establish immortalized embryonic fibroblast lines in heat shock transcription factor 1 (HSF1) HSF1-/- and HSF1+/+ mice and to provide experimental models to study the function of HSF1. METHODS: A mammalian expression vector (pSV3neo) containing the SV40 large T antigen was used to transfect the HSF1-/- and HSF1+/+ mouse embryonic fibroblast using Lipofectamine 2000. Colonies were screened by G418 and expanded to immortalized cell lines. PCR was used to detect the integration of the large T antigen with genome in the mouse embryonic fibroblast. Expression of SV40 large T antigen gene in expanded cells was identified by RT-PCR. HSP70 expression was examined by Western blot in the embryonic fibroblast lines. RESULTS: The stable growth and serial propagation were observed in the HSF1-/- and HSF1+/+ cell lines for six months. The mRNA of SV40 T antigen gene expressed in the two cell lines. HSP70 expression could not be induced in the heat-treated HSF1-/- mouse embryo fibroblasts. CONCLUSION: The immortalized cells of HSF1+/+ and HSF1-/- mouse embryo fibroblasts are successfully established. PMID- 16706110 TI - [Protective effect of bombesin receptor subtype-3 on human brochial epithelial cells against injury]. AB - OBJECTIVE: To investigate the role and mechanism of bombesin receptor subtype 3 (BRS-3) in the proliferation and protection against injury of human brochial epithelial cells (HBECs). METHODS: Effect of P3513 (a specific agonist of BRS-3) on the proliferation of HBECs was observed by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) method; the release rate of 3H-Udr, and LDH activity, catalase activity, and the expression of cadherin and integrin beta1 were also analyzed under O3 stress with or without P3513 treatment. RESULTS: The proliferation of HBECs was accelerated by P3513 in a concentration-dependent manner (10(-9) approximately 10(-7) mol/L). Ozone stress could promote the release rate of 3H-Udr, and LDH activity, which could be inhibited by P3513. P3513 could promote the activity of catalase. The effect of proliferation and protection against injury caused by P3513 could be inhibited by W7 (calmodulin inhibitor), PD98059 (tyrosin kinase inhibitor) and H89 (PKA inhibitor). P3513 could stimulate the expression of caderin and integrinbeta1 of ozone-stressed HBECs. CONCLUSION: Activation of BRS-3 caused by P3513 may play an important role in protecting HBECs from oxidant injury, and the signal pathway is possibly relevant to Ca2+, MEK and PKA. PMID- 16706111 TI - [Chinese version of SF-36 in the quality of life assessment among community dwelling elders]. AB - OBJECTIVE: To obtain scores in a community-dwelling population over 60 using the SF-36, to assess the reliability and validity of this general health questionnaire, and to analyze the difference in dimension scores among the elderly Chinese in Changsha. METHODS: We randomly selected 602 elders, aged 60 to 91 years, in multi-phases. All the subjects had resided in Changsha for at least one year. The reliability of the SF-36 was assessed by split-half reliability and Cronbach's alpha coefficient and the validity through factor analysis and correlation analysis, etc. The dimension scores of different people were obtained by analysis of variance and independent-samples t-test. RESULTS: The split-half reliability was 0.72 and the Cronbach's alpha coefficients of all the 8 dimensions were more than 0.8; the Pearson correlate coefficients of all the items to their dimensions were more than 0. 59. SF-36 contained 8 domains and 2 summary scales in the factor analysis. Health-related quality was different in different elders. CONCLUSION: The SF-36 is practical in studying the quality of life among community-dwelling elders. PMID- 16706112 TI - [Effect of health education on the lung function and life quality in patients with stable chronic obstructive pulmonary diseases]. AB - OBJECTIVE: To evaluate the effect of health education on the symptoms, lung function and life quality in patients with stable chronic obstructive pulmonary diseases (COPD). METHODS: Eighty-two patients were assigned into 2 groups randomly: The treatment group (n = 43) accepted health education for 6 months and the control group (n = 39) did not. Before and after the 6 months, we observed the number of smokers, Borg score, inhale treatment, times of acute episode, SGRQ score, lung function, and the therapeutic effect in the two groups. RESULTS: Six months later, the ratio of smokers in the treatment group was 13.95%, much lower than that in the control group (35.90%, P = 0.021); the Borg score of the treatment group decreased from 4.86 +/- 1.21 to 3.38 +/- 0.94 (P =0.000), but there was no difference in the control group; the ratios of accepted inhale treatment in the treatment group and the control group were 100% (43/43) and 20.51 % (8/39) respectively (P = 0.000), and the accuracy rates were 100% (43/43 ) in the treatment group and 12. 82% (5/39) in the control group (P =0. 000); the times of acute episode in the treatment group was 1.51 +/- 1.53, much lower than that in the control group (4.46 +/- 5.17, P = 0. 000); the indexes of lung function before and after the 6 months between the two groups had no significant difference. The total score, the symptom section score, the activity section score, and the impact section score of SGRQ after the 6 months of health education were much lower than those of 6 months before, and those in the control group had no significant difference. CONCLUSION: Six months of the health education can decrease the ratio of smokers and the times of acute episode, and to improve the life quality of patients with stable COPD. PMID- 16706113 TI - [Effect of polygonum multiflorum thunb on BDNF expression in rat hippocampus induced by amyloid beta-protein (Abeta) 1-40]. AB - OBJECTIVE: To explore the effect of polygonum multiflorum thunb (PMT) on the expression of brain-drived neurtrophic factor (BDNF) in hippocampus CA1 in rats induced by Abeta1-40. METHODS: The rat model was established by injecting Abeta1 40 into the right dorsal hippocampus in rats, and their behavior was observed by Y-maze. The morphological changes of rat hippocampus were examined by the Nissl staining. The expression of BDNF in hippocampus CA1 neurons was also examined by immunohistochemistry and Western blot. RESULTS: After the administration of Abeta1-40, the times of rats learning how to elude electric stimulation were significantly increased, and the expression of BDNF in hippocampus CA1 were obviously reduced. The aggregation of Abeta1-40 in hippocampus and the morphological changes of CA1 hippocampus were found. After the treatment with PMT for 30 days, the times learning how to elude electric stimulation were obviously reduced, while the expression of BDNF in hippocampus CA1 was also increased. CONCLUSION: PMT can reverse the down-regulation of BDNF expression in hippocampus CA1 induced by Abeta1-40. PMID- 16706114 TI - [Detection of MSH2 gene mutation by PCR]. AB - OBJECTIVE: To establish a genetic diagnosis method for a novel MSH2 mutation. METHODS: A specific primer on the mutated site of MSH2 was synthesized and PCR was conducted using the specific primer and another downstream primer. PCR products were electrophoresed and then the carriers with the novel gene mutation of the carriers or non-carriers were identified. RESULTS: MSH2 in a hereditary nonpolyposis colorectal cancer family were successfully found. CONCLUSION: The method is effective and simple for genetic diagnosis of the novel mutation in MSH2. PMID- 16706115 TI - [Effect of hematopoietic stimulating factors on the expansion of megakaryocyte]. AB - OBJECTIVE: To investigate the effect of hematopoietic stimulating factors on the expansion of mature megakaryocytes. METHODS: (2, 4, 6, 8, 10) x 10(5)/mL bone marrow single nucleus cells (BMNC) were added in the culture system of colony forming unit-megkaryocyte (CFU-Meg) to find out the relationship of the cultured BMNC with the output of CFU-Meg. rmSCF + rmTPO + rmIL-3 (3HSFs) and rmSCF + rmTPO + rmIL-3 + rmIL-6 (4HSFs) or F-CM were added in the liquid culture system of megkaryocytes respectively. The number of mature megakaryocytes were counted every other day. RESULTS: The number of CFU-Meg increased with the increase of the cultured BMNC. The CFU-Meg productivity of 1 x 10(6) BMNC/mL culture system was more than that of 2 x 10(5) BMNC/mL culture system. 3HSFs and 4HSFs or F-CM significantly promoted the expansion of mature megakaryocytes in the liquid culture system, but the effect was different. The peak time of the number of mature megakaryocytes in 3HSFs and 4HSFs or F-CM were 7 d, 7 d and 5 d respectively. CONCLUSION: 3HSFs and 4 HSFs or F-CM had positive effect on the expansion of mature megakaryocytes. 4HSFs was better than 3HSFs and F-CM. 3HSFs was better than F-CM. The peak time of the number of mature megakaryocytes in different culture systems was different. PMID- 16706116 TI - [Causes and revision endoscopic sinus surgery for recurrent sinusitis]. AB - OBJECTIVE: To investigate the causes of revision endoscopic sinus surgery (RESS), and to evaluate the clinical effect and experience of RESS in the patients with recurrent sinusitis. METHODS: Before the revision surgery, 168 patients (225 sides) with recurrent sinusitis were examed by CT scans and nasal endoscopy. All patients were operated by revision endoscopic sinus surgery. RESULTS: Among the 168 patients, 95 cases (121 sides) had incomplete middle turbinate, 51 cases (75 sides) had maxillary sinus ostium stenosis, 85 cases (117 sides) had nasal cavity adhesion, 48 cases (57 sides) had uncinate process residual, 38 cases (65 sides) had ethmoid hyperostosis, and 25 cases had nasal septum deviation. One hundred and six patients (134 sides) were cured, 40 patients (56 sides) were improved, useful 22 patients (35 sides) were ineffective. CONCLUSION: The most common surgical causes of failures in endoscopic sinus surgery are the disease area remnant, nasal cavity adhesion, maxillary sinus ostium stenosis, uncinate process residual, ethmoid hyperostosis, and nasal septum deviation. Revision endoscopic sinus surgery is a useful way to treat recurrent sinusitis. Preoperative CT scan, correct choice of the anatomic marks of orientation and direction function are the key to operation successful. PMID- 16706117 TI - [Callotasis for segmental bone defects in the femur]. AB - OBJECTIVE: To study the clinical efficacy of callotasis for pathological segmental defects in child femur. METHODS: Thirty-nine patients with segmental femoral defects suffered from suppurative osteomyelitis were treated with the bilateral, unilateral external fixation frame or intramedullary callotasis. The rolongational rate was from 2 to 2.5 cm per month after the operation. RESULTS: After a 13 to 57 month follow-up, all cases were healed according to clinical examination and radiological observation. The prolongational length ranged from 9 to 31 centimeters. Average ratio of prolongation reached 49%. The healing index was 30 d/cm, and the healing time of non-union was 134 days. No recurrent suppurative osteomyelitis or fracture was observed. CONCLUSION: Callotasis is a reliable, simple and hyperadaptable method for femural segmental defects. PMID- 16706118 TI - [Effect of surgically induced bilateral cleft alveolar on maxillary growth in dogs]. AB - OBJECTIVE: To establish an animal model of bilateral alveolar cleft and to determine the effect of cleft on maxillary growth. METHODS: Eight dogs (12 weeks old) were divided into unoperated control groups (n = 4) and the model group (n = 4). The model dogs were operated to establish a bilateral alveolar cleft. All the dogs were killed and the craniofacial morphology on clean skull was analyzed by the direct detection and CT. RESULTS: The length, the foreside width, and the foreside height of the maxillary in the model group were shrunken compared with those of the control group (P < 0.05). CONCLUSION: The alveolar cleft plays an important role in the maxillary aberration. PMID- 16706119 TI - [Effect of androgen on nitric oxiode synthase activity in the kidney of rats with ischemic injury]. AB - OBJECTIVE: To investigate the effect of androgen on nitric oxide synthase activity and kidney function in the kidney of rats with ischemic injury. METHODS: Fourty-eight adult healthy male SD rats were randomly divided into the renal ischemia group (n = 24 ) and the orchidectomized group with renal ischemia (n = 24). The two groups were randomly divided into 4 groups by the survival time: 0, 1, 3 and 7 days. NADPH diaphorase histochemistry was used to measure the nitric oxide synthase activity of the kidney with ischemic injury in the orchidectomized group and the control group. The image analysis technology system was used to quantify the NOS. BUN and BCr in the plasma were detected when the rats were executed. RESULTS: The NOS histochemical staining of the kidney was mainly distributed in the renal tubule, macula densa and low in the renal glomerular. The NOS activity of macula densa significantly decreased in the orchidectomized group compared with that in the control group (P < 0.05) . BUN and Bcr have no significant difference on 0, 1 and 7 days (P > 0.05) , but on the third day the data markedly increased (P < 0.05). CONCLUSION: Androgen can adjust the blood flux of glomerular by regulating the NOS activity of macula densa, and affect the kidney function. PMID- 16706120 TI - [Specific cell immune response mediated by dendritic cells in multiple myeloma patients in vitro]. AB - OBJECTIVE: To explore KM3 multiple myeloma (MM) cell line specific cytotoxic T lymphocyte (CTL) response in vitro mediated by autologous dendritic cells (DCs) aroused by KM3 cells' lysates and acid-eluted peptides. METHODS: Monocytes were isolated from the peripheral blood of healthy individuals and MM patients, and were cultured in serum medium with IL-4, GM-CSF, and TNF-alpha to generate DCs. These DCs were pulsed by KM3 cells' lysates or the acid-eluted peptides, then incubated with autologous T lymphocytes for 3 days to induce KM3 cell antigen specific CTL. MTT assay was performed to examine the specific KM3 cells' lysing ability of CTL. RESULTS: DCs were generated in peripheral blood monocytes cultured in the serum medium containing GM-CSF, IL-4, and TNF-alpha. Autologous T lymphocytes induced by IL-2 and DCs pulsed with KM3 cells' lysates or the acid eluted peptides showed strong killing effect on KM3 cells. CONCLUSION: The DCs pulsed by KM3 cells' lysates or the acid-eluted peptides incubated with autologous T lymphocytes can induce KM3 cell antigen specific CTL. PMID- 16706121 TI - [Effect of HSP72 on acute injury of neonatal cardiomyocytes induced by oxidative stress]. AB - OBJECTIVE: To explore the protective effect of HSP72 on the acute injury of cardiomyocyte induced by oxidative stress. METHODS: Cardiomyocytes of neonatal rats treated with heat shock (42 degrees C, 30 min, recovery for 6 h) to induce the expression of HSP72 and HSP72 antisense oligonucleotide was transformed to block the expression of HSP72. 0.5 mmol/L (final concentration) H2O2 was added into the culture medium to mimic oxidative stress, and to induce the acute injury of neonatal cardiomyocytes. The release of LDH and the total protein synthesis were applied to evaluate the injury of cardiomyocyte of neonatal rats. RESULTS: Oxidative stress could significantly increase the release of LDH, and inhibit the total protein synthesis. By inducing the expression of HSPs, heat shock pretreatment significantly reduced the release of LDH and relieved the oxidative stress-mediated inhibition of total protein synthesis. Moreover, HSP7-2 anti sense oligonucleotide could remarkably block the protective effect of heat shock pretreatment on the cellular injuries induced by H2O2. CONCLUSION: HSP72 plays a most important role in the acute injury of cardiomyocyte mediated by oxidative stress. PMID- 16706122 TI - [Variations of beta-cell early-phase insulin secretion in type 2 diabetic patients in different stages]. AB - OBJECTIVE: To explore the variations of early-phase insulin secretion in Type 2 diabetic patients in different stages. METHODS: L-arginine stimulative test, fast blood glucose and body mass index (BMI) were evaluated in 40 nomal controls (NC) and 101 Type 2 diabetic patients. The diabetic patients were divided into three groups: newly diagnosed group (n = 35), effectively treated by sulfonylureas group (n = 32) , and secondary failure of sulfonylureas group (n = 34). The indexs of insulin resistance of homeostasis model assessment (HOMA-IR), beta-cell insulin secretion of homeostasis model assessment (HOMA-IS), and the acute insulin response (AIRARG) index were calculated. Some statistical comparisons were done among the 4 groups. RESULTS: The indexs of HOMA-IR in each group of Type 2 diabetic patients were all higher than those in NC group (P < 0.01). The AIRARG indexs were obviously lower in Type 2 diabetic patients in different stages than those in NC group (P < 0.01), and the subsequence from the highest to the lowest among the groups of diabetic patients was: the newly diagnosed group, the effectively treated by sulfonylureas group, and the secondary failure of sulfonylureas group (P < 0.01). But there was no significant difference in indexs of HOMA-IS between the newly diagnosed group and the effectively treated by sulfonylureas group. CONCLUSION: There is severe insulin resistance in Type 2 diabetic patients in each stage. The variations of early-phase insulin secretion manifest a vary procedure of obvious deterioration by degrees from the newly diagnosed group to the secondary failure of sulfonylureas group in Type 2 diabetic patients. PMID- 16706123 TI - [Apoptosis and c-myc protein expression in the retinal of form-deprivation myopia]. AB - OBJECTIVE: To study the apoptosis of retina and the expression of c-myc protein in form-deprivation myopia. METHODS: Two-day-old chickens were sutured with right eyelid for 4, 8 and 12 weeks. After measurement of refracation, the eyeballs were observed by light microscope and taken photos. Retinal apoptotic cells were measured by TUNEL staining and flow cytometry. C-myc protein were examined by immunohistochemistry and flow cytometry. RESULTS: Lacquer crack lesions were found in sutured eyes at 12 weeks. Apoptotic cells were observed in retinal outer and inner nuclear layer of the sutured eyes at 12 weeks and obvious peak of apoptosis was observed in sutured eyes at 12 weeks. The expression of c-myc protein was significantly more than control eyes at 8 and 12 weeks. CONCLUSION: The apoptosis of retinal was present in form-deprivation myopia with the degeneration of retina. C-myc protein plays an important role in retinal apoptosis of myopia. PMID- 16706124 TI - [Protective effect of lidocaine on injury alveolar Type II cells induced by LPS in adult rats]. AB - OBJECTIVE: To investigate the effect of lidocaine on LPS induced apoptosis of cultured adult rat alveolar Type II (AT-II) cells. METHODS: Cultured cells were exposed to LPS and lidocaine for 24 hours. Apoptosis and necrosis rates of cells were detected by flow cytometry and electron microscope. The activity of lactic dehydrogenase (LDH) was analyzed by using LDH kits. RESULTS: LPS induced the AT II cell injuries by increasing not only the necrosis and apoptosis rates but also the LDH release of cultured AT-II in vitro. Lidocaine decreased the necrosis and apoptosis rates of AT-II cells. CONCLUSION: Lidocaine can directly inhibit the apoptosis and necrosis induced by LPS in cultured AT-II cells. PMID- 16706125 TI - [Effect of chloroquine on the apoptosis of intestinal mucosa epithelial cells and enterogenous bacteria-endotoxin translocation after total hepatic ischemia reperfusion in rats]. AB - OBJECTIVE: To observe the effect of chloroquine on the apoptosis of intestinal mucosa epithelial cell and enterogenous bacteria-endotoxin translocation after total hepatic ischemia-reperfusion in rats. METHODS: The rat total hepatic ischemia-reperfusion model was built by blocking the hepatic portal, suprahepatic and infrahepatic vena cava for 20 minutes. Ninety Sprague-Dawley rats were assigned randomly into the sham operation group (Group A, n = 30), total hepatic ischemia-reperfusion treatment group (Group B, n = 30), and chloroquine administrated group (Group C, n = 30). Each group was subdivided randomly into 3 subgroups (n = 10) according to different experiment time phases as follows: after 20 minutes of total hepatic vascular exclusion (T0), 4 hours after reperfusion (T1), and the 48 hours of survival. Group A and Group B were intravenously injected with normal saline 1 mL/kg while Group C received chloroquine 10 mg/kg which dissolved in 1 mL/kg normal saline intravenously. The levels of portal blood D-lactate, TNF-alpha, endotoxin, and the intestinal mucosa MDA concentration were measured at T0 and T1; the portal blood, mesenteric lymph node, and spleen tissues were cultured for bacteria; and the apoptotic index of intestinal mucosa epithelial cells at T0 and T1 and the survival rate after 48 hour reperfusion were obtained. RESULTS: Compared with Group A, the levels of portal blood D-lactate, TNF-alpha, endotoxin and the intestinal mucosa MDA in Group B and Group C were significantly higher (P < 0.05 or P < 0.01). These indexes of Group C were lower than those of Group B (P < 0.05). The portal vein blood, mesenteric lymph node and spleen tissues existed the bacterium translocation both in Group B and Group C, and the positive rate in Group C was lower than that in Group B (P < 0.05). Apoptotic index of the intestinal mucosa epithelial cell increased significantly in Group B (P < 0.01) and Group C (P < 0.05), but the apoptotic index in Group C was lower than that in Group B (P < 0.05); the 48 hour survival rate of the rats in Group C was higher than that in group B (P < 0.05). CONCLUSION: Chloroquine may decrease the intestinal mucosa epithelial cell apoptosis and the enterogenous bacteria-endotoxin translocation after total hepatic ischemia-reperfusion and increase the survival rate of the rats. PMID- 16706126 TI - [Influences of perioperative metoprolol on hemodynamics and myocardial ischaemia in elderly patients undergoing noncardiac surgery]. AB - OBJECTIVE: To observe the influences of metoprolol on hemodynamics and myocardial ischaemia in elderly patients undergoing noncardiac surgery. METHODS: Thrity patients (60 approximately 75 years) undergoing elective noncardiac surgery were randomly divided into a metoprolol group (n = 15) and a control group (n = 15). In the metoprolol group, metoprolol (0. 5 mg and 1.5 mg) was slowly injected into the vein of patients before the induction of intravenous anesthesia and after the tracheal intubation. The hemodynamic indice (invasive BP, HR and rate pressure product-RPP), the myocardial ischaemia indice (reversible ST segment depression of ECG II, V5 leads more than 0.1 mv or reversible ST segment elevation more than 0.2 mv from the baseline, ST segment depression or elevation over 1 min), the myocardial damage indice (serum cardiac troponin I, cTn I), and the indice of metoprolol cardiac and the respiratory adverse effects (incidence of HR below 50 beats/min, average doses of atropine, airway peak pressure) were observed intraoperatively. RESULTS: The HR and RPP were lower before the tracheal induction than the baseline (before anesthesia) in all patients, but there is no significant difference between the two groups (P > 0.05). During the tracheal intubation, the HR and RPP of the control group significantly increased, compared with the baseline (P < 0.05) and those of metoprolol group (P < 0.05). The incidence of perioperative hypertension was higher in the control group than that in the metoprolol group. The incidence of myocardical ischaemia episode was 30% in the control group, and 13% in the metoprolol group (P < 0.01). The release of cTn I was detected in 5 patients in the control group, and 2 patients in the metoprolol group (P < 0.05). The incidence of HR < 50 beats/min, and the average doses of atropine had no statistic difference between the two groups, but a tendency of high incidence of bradycardia in the metoprolol group occurred when abdominal viscera was tracted by surgical manupilation. There was no significant difference in airway peak pressure, SpO2 and PET CO2 between the two groups (P > 0.05). CONCLUSION: Intravenous administration of 0.5 mg and 1. 5 mg metoprolol before the induction of anesthesia and after the tracheal intubation has several advantages, including the decrease of myocardial oxygen consumption, the improvement of hemodynamic stability, and the lowering perioperative incidence of myocardial ischeamia and damage, but the tendency of high bradycardia incidence caused by peritoneal traction should be noticed. PMID- 16706127 TI - [Effect of probucol and losartan on growth factors after balloon angioplasty in rabbits]. AB - OBJECTIVE: To investigate the effect of probucol and losartan on the prevention of restenosis after balloon angioplasty in hypercholesterolaemic rabbits, and to examine the expression of growth factors. METHODS: Forty male New Zealand white rabbits were randomly divided into high cholesterol diet group, probucol group, losartan group and combined drugs group. After one week of diet, all rabbits were injured on iliac arteries with balloon. Four weeks after the injury, the morphology of the iliac arteries of the rabbits were observed, and the insulin like growth factor-I receptor (IGF-IR) and vascular endothelial growth factor (VEGF) were examined by immunohistochemical methods. RESULTS: Compared with the high cholesterol diet group, the lumen areas of the probucol group, losartan group and combined drugs group were larger (P < 0.01), the intimal areas were smaller (P < 0.05), and the expression of IGF-IR and VEGF significantly decreased (P < 0.05). There was no statistically significant difference among the three groups. CONCLUSION: Probucol and losartan can prevent the restenosis of rabbits' iliac artery from balloon injury, and inhibit the expression of IGF-IR and VEGF. There is no statistical difference between combined drugs and single drug administration. PMID- 16706128 TI - [Measurement peripheral blood index related to inflammation and ox-LDL, ox-LDLAb in patients with coronary heart disease and its clinical significance]. AB - OBJECTIVE: To detect the levels of index related to inflammation such as soluble CD40 ligand (sCD40L), neutrophil collagenase-8 (MMP-8), and pregnancy associated plasma protein-A (PAPP-A) , lipid peroxidation and autoimmune indexes such as oxidized low density lipoprotein (ox-LDL) and its antibody (ox-LDL Ab) in patients with coronary heart disease, and to investigate its relationship with acute coronary syndrome (ACS). METHODS: Contents of sCD40L, MMP-8, PAPP-A, ox-LDL and ox-LDL Ab in the peripheral blood were measured by enzyme-linked immunosorbent assay from 109 patients with coronary heart disease including 36 acute myocardial infarction (AMI), 38 unstable angina pectoris (UAP), and 35 stable angina pectoris (SAP) and 36 controls without coronary heart disease. RESULTS: The levels of each index in the peripheral blood of ACS patients (including AMI and UAP) were higher than those of SAP patients and controls (P < 0.05), and the difference of each index between UAP group and AMI group in ACS patients had no statistical significance (P > 0.05). The levels of each index of SAP patients, except PAPP-A, were all higher than those of controls (P <0.05). All the indexes were helpful in diagnosis of ACS. The area under the ROC curve of each index is between 0.7 and 0.9. CONCLUSION: The increase of sCD40L, MMP-8, PAPP-A, ox-LDL and ox-LDL Ab levels in peripheral blood may be related to the pathogenesis of ACS, and can be used as potential markers of unstable atherosclerosis plaque. PMID- 16706129 TI - [Evolution rules and clinical value of apparent diffusion coefficient in cerebral infarction]. AB - OBJECTIVE: To determine the rules that apparent diffusion coefficient (ADC) changes with time and space in cerebral infarction, and to provide the evidence in defining the stage, guiding treatment or judging the prognosis in infarction. METHODS: Eighty-eight work-ups in 69 patients with cerebral infarction (8 hyperacute, 35 acute, 20 subacute, and 25 chronic infarctions) were imaged with both conventional MRI and diffusion weighted imaging. The average ADC, the average relative ADC (rADC), and the ADC or rADC from the center to the periphery of the lesion were calculated. RESULTS: The average ADC and the average rADC of hyperacute and acute infarction lesion depressed obviously. The average ADC and the average rADC of subacute infarction lesion were significantly higher than those of hyperacute and acute infarction lesion (P < 0.05), and some approached the values of uninjured side which appeared "pseudonormal values" at 10 to 14 days. The average ADC and the average rADC in chronic infarction lesion were the highest (P < 0.05). The longer the prognosis, the higher the average ADC and the average rADC of infarction lesion. The ADC and the rADC in 7 hyperacute and 26 acute lesions had gradient signs that these lesions increased from the center to the periphery. The ADC and the rADC in 15 subacute lesions had adverse gradient signs that these lesions decreased from the center to the periphery. CONCLUSION: The ADC and the rADC of infarction lesions have evolution rules with time and space corresponding to the pathophysiological foundation. The evolution rules with time and those in space can be helpful to decide the clinical stage and pathophysiological phase of infarction, and to provide the evidence in guiding the treatment or judging the prognosis in infarction. PMID- 16706130 TI - [Detection and clinical significance of urinary epidermal growth factor in brain tumor patients]. AB - OBJECTIVE: To investigate the clinical significance of urinary epidermal growth factor (EGF) in patients with brain tumors. METHODS: The levels of EGF in urine samples collected from 20 patients (9 low grade astrocytomas, 6 anaplastic astrocytomas, and 5 meningiomas) and 5 healthy individuals were determined. EGF levels were measured by radioimmunoassay technique. A preoperative and one postoperative determination were performed. RESULTS: Preoperative urinary EGF levels of astrocytoma patients were statistically higher than those of meningioma patients and the controls (P < 0.01). Preoperative urinary EGF levels showed a positive correlation with the degree of malignance in the astrocytoma patients (P < 0.05). A significant decrease of the postoperative levels of EGF was observed in the astrocytoma patients who underwent gross total resection (P < 0.01). The pre/postoperative urinary EGF levels of the meningioma patients showed no significant fluctuations and showed no significant difference with those of healthy individuals (P > 0.05). CONCLUSION: The urinary EGF levels of astrocytoma patients correlate with the WHO grade of malignance and significantly decrease after gross total removal. Urinary EGF may be of practical value in diagnosing and evaluating the surgical efficacy of astrocytomas. PMID- 16706131 TI - [Clinical analysis of pregnancy complicated with systemic lupus erythematosus]. AB - OBJECTIVE: To investigate the interacting effects between pregnancy and flares of systemic lupus erythematosus (SLE) and to explore the best occasion for SLE patients' conception and the management during the pregnancy. METHODS: Thirty one cases of pregnancy complicated with SLE were investigated retrospectively, among whom 18 were in remission of SLE at the beginning of conception (Group A), and the other 13 either had high-activity of the disease or were first diagnosed as SLE during the pregnancy (Group B). Various doses of prednisone were administered to control SLE. RESULTS: SLE flares still occurred in 6 cases in Group A, but in all cases in Group B. Compared with Group A, the rates of fetal loss and early delivery were significantly higher in Group B (P < 0.05), while the survival rate and the weight of the new born were notably decreased in Group B (P < 0.05). CONCLUSION: Pregnancy and SLE interacted with each other unfavorably. Selection of remission stage for conception and proper management during the pregnancy could significantly improve the maternal-fetal safety. PMID- 16706132 TI - [Pathologic features and prognosis of 21 children with isolated proteinuria]. AB - OBJECTIVE: To discuss the pathologic features, treatment and prognosis of the children with isolated proteinuria (IP). METHODS: Twenty-one children with IP were enrolled according to their renal biopsy and were followed up for 0.5 to 10 years. RESULTS: Renal biopsy was performed in all children. Among them 13 were mesangial proliferation glomerulonephritis (MsPGN) (including 3 minor, 6 moderate, and 4 severe ones), 2 minimal change nephritis (MCN), 3 IgA nephropathy (IgAN) (1 in Grade I and 2 in Grade II), 2 focal segmemtal glomerulosclerosis (FSGS) and 1 endocapillary proliferative glomerulonephritis (EnPGN). Interstitial changes could be found in MsPGN and FSGS mostly, presenting interstitial fibrosis, infiltration of inflammatory cells, atrophy of renal tubule, and the vacuolar degeneration of epithelia. All children accepted the medical treatment except the EnPGN case. Fifteen children recovered with no relapse; proteinuria persisted in 3 severe MsPGN and FSGS cases; 2 got the impaired renal function accompanied by persistent proteinuria; and 1 had hypertension. CONCLUSION: The different degrees of renal damage can be found in all IP children who have persistent proteinuria. Most patients can get good outcome after aggressive therapies. However, the prognosis of those with severe MsPGN and FSGS was not so optimistic, and some reno-protective treatments should be given to postpone the deterioration of the renal function. PMID- 16706133 TI - [Distribution of pathogen and resistance of nosocomial infections in the intensive care units]. AB - OBJECTIVE: To determine the distribution of pathogens and their characteristics of drug susceptibility originating from nosocomial infections in the intensive care units (ICU), and to provide evidence for clinical anti-infection treatments. METHODS: Retrospective analysis to the pathogens and their drug susceptibility characteristics was carried out. These pathogens were isolated from the samples that came from patients infected in the ICU from 2002 to 2004. RESULTS: The main nosocomial infective pathogens in the ICU were gram negative bacilli (48.2%), and the next ones were gram positive bacteria (43.3%) and fungus (8.5%). The most common gram negative bacilli were Pseudomonas aeruginosa and Escherichia coli; while for gram positive bacteria, the main bacterin were Staphylococcus aureus. The gram negative bacilli could resist 4 or more than 4 antibiotics, and the rate for resistance exceeded 40%. Similarly, oxacillin resistance staphylococcus could resist 7 antibiotics, and the rate was over 50%. The detective rates of ESBLs and AmpC enzymes produced by Escherichia coli and K. peumoniae were 34.0% & 30.7% and 13.2% & 23.1%, respectively. The rate for oxacillin resistance staphylococcus was 66.3%, and there was relative high resistance rate ( > 55%) for most antibiotics: There was statistical difference, compared with that of non-resistant strains. CONCLUSION: Though gram positive coccus still play an important role, most infections are caused by gram negative bacilli of nosocomial infections in the ICU. The antibiotics resistant rate of all bacteria has been rising gradually. It shows strong resistance and multi-drug resistance. The most importment cause for resistance of gram negative bacilli is that the bacteria can produce ESBLs and AmpC enzymes. The antibiotic resistant rate for oxacillin resistance staphylococcus is really high. PMID- 16706134 TI - [Mononostril-septum-transsphenoidal approach for pituitary adenoma]. AB - OBJECTIVE: To summarize the mononostril-septum-transsphenoidal approach for pituitary adenoma. METHODS: The clinical features, operative techniques, and outcome of 36 patients with pituitary adenoma were analyzed retrospectively. RESULTS: Tumors were totally removed in 28 cases, and subtotally resected in 8 patients. No patient died after the operation. Endocrine symptom of 31 patients returned to the normal level, the symptom of the other 5 cases were improved. Thirty patients with visual field defects recovered after the operation. Cerebrospinal fluid leakage occurred in one patient, and was cured with conservative treatment in 2 weeks. CONCLUSION: Mononostril-septum-transsphenoidal approach can make use of the natural space of the nasal cavity, which has many advantages, such as direct approach, short operative time, minimal invasion, and few complications. It is a effective transsphenoidal surgical approach. PMID- 16706135 TI - [Clinical application of video-assisted thoracoscopic surgery]. AB - OBJECTIVE: To explore the clinical application of video-assisted thoracoscopic surgery (VATS). METHODS: We retrospectively analyzed the clinical data of 672 cases of VATS. There were 17 thoracic diseases such as emphysema, bullectomy for spontaneous pneumothorax, massive bullae, benign tumor of mediastinum, cyst of mediastinum, pulmonary benign tumors, hydropericardium, malignant pleural fluid, etc. RESULTS: The mean operation time was 57 minutes and there were no intraoperative complications. The bleeding during the operation was less than 100 mL. Postoperative pneumothorax occurred in 4 patients and among them 2 patients were of relapse after 1 month. The intrathoracic drain in most patients was removed with an average of 2. 5 days. A supplementary incision was needed in 10 cases: Six were due to the adhesion of full pleural cavity and 4 were found with the malignant tumor during the operation. CONCLUSION: VATS is an alternative approach that provides a safe, less invasive, and effective operation for treating spontaneous pneumothorax, benign tumor of mediastinum, cyst of mediastinum, pulmonary benign tumors, pericardial perfusion, and acute chest trauma patients. PMID- 16706136 TI - [Screw-rod technique in treating cervical spinal cord injury without fracture and dislocation]. AB - OBJECTIVE: To evaluate the efficacy of screw-rod technique in treating cervical spinal cord injury without fracture and dislocation. METHODS: The screw-rod technique was used to treat 38 patients with cervical spinal cord injury without fracture and dislocation. The JOA scores before and after the surgery were evaluated. RESULTS: No severe complication occurred. The JOA scores were increased by 4.1, 6.7, 7.8, and 8.1 respectively at 0 th, 4 th, 12 th and 24 th weeks after the operation. Neurological deficits were improved one year after the operation in 1 patient. CONCLUSION: Treating cervical spinal cord injury without fracture and dislocation with screw-rod technique can achive an efficient and safe clinical outcome. PMID- 16706137 TI - [Treatment of spinal metastases by posterolateral approach]. AB - OBJECTIVE: To evaluate the value of spondylectomy, circumferential decompression, and spinal reconstruction for the treatment of spinal metastases using posterolater approach. METHODS: Thirteen patients with spinal metastases were treated by spondylectomy, circumferential spinal cord decompression, and spinal reconstruction with pedicle fixation and polymethyl methacrylate. The outcome was assessed by pain relief and the improvement in neurologic symptoms. RESULTS: Pain relief was obvious in all patients. Neurologic symptoms improved to various degrees in 6 patients. CONCLUSION: Spondylectomy, spinal cord circumferential decompression and spinal reconstruction with pedicle fixation and polymethyl methacrylate using posterolateral approach are effective surgical method for spinal metastases. PMID- 16706138 TI - [Interventional treatment for 2 cases of congenital heart disease associated with defect of hepato-department of inferior vein cava]. PMID- 16706139 TI - [Benign pancreatic diseases treated with spleen-preserving distal pancreatectomy with conservation of the splenic vessels]. PMID- 16706140 TI - [Common mistakes and nonstandard expressions in English abstracts of medical literatures]. AB - The mistakes in English abstracts of medical literatures influence the international communication of medical information. The misuse of words, grammar mistakes, and improper expressions are common problems. Moreover, nonstandard expressions also include the names of Chinese herbal medicines, numbers, punctuations, and so on. PMID- 16706141 TI - [Cellular mechanisms of cirrhotic rat liver regeneration. II. Proliferation, polyploidization and hypertrophy after partial hepatectomy]. AB - Using cytofluorimetry and absorptional cytophotometry, hepatocyte DNA and total protein contents were measured in intact and cirrhotic rats in 1, 3 and 6 months after partial hepatectomy (PH). It has been found that within one month of intact rat liver regeneration the level of hepatocyte ploidy rised by 25% to remain elevated for the next 6 months. This was due mainly to reducing the number of cells with diploid nuclei (2c 2-fold, 2c x 2 - 6.6-fold) and to rising the number of octaploid hepatocytes. In cirrhotic animals the ploidy level in hepatocytes increased in 3 months after PH, and decreased by 15% in 6 months. The number of hepatocytes with diploid nuclei (2c and 2c x 2) increased within 3-6 months in both control and cirrhotic rats. The protein content per diploid hepatocyte rised by 30% within 3-6 months of liver regeneration after PH. Special calculations have shown that within 3 months after PH the increase in the liver mass of control and cirrhotic rats was due completely to hepatocyte DNA synthesis, i. e. proliferation and polyploidization. Within the next 3 months of liver regeneration after PH, the contribution of polyploidization to liver mass increase was negative because of depolyploidization of liver parenchyma cell population. At this time hypertrophy was the main process determining the liver mass increase. PMID- 16706142 TI - [Change in synthetic activity of epidermis cells in rats during burn wound healing under a scab and in liquid environment]. AB - One of the problems of burn treatment is a creation of conditions providing most valuable skin rehabilitation. An experimental model of burn wound healing in a 0.9% NaCl solution is proposed. Synthetic activity of rat epidermis cells in the process of burn wound healing under a scab and in liquid environment was studied by luminescent microscopy. The effect of a 0.9% NaCl solution involves an increase of the basal layer cell synthetic activity of regenerating epidermis, and keeping a high level of this activity of hair follicle epithelial cells for a long time. Tissue-preserving effect of the 0.9% NaCl solution on burns healing has been confirmed in these results. PMID- 16706143 TI - [Proliferative activity of human endothelial cell line EA.hy926 and its modulation]. AB - Angiogenesis is a complicated process, which is regulated by numerous cytokines and growth factors. Besides, the interaction of endothelial cells with extracellular matrix components, with other cell types and with each other is essential for the formation on new blood vessels. The initiation, continuation and completion of angiogenesis depend on the balance of pro- and antiangiogenic factors in the endothelium microenvironment. Factors that influence endothelial cell proliferation are necessary for vascular development and their normal functioning. The influence of new pharmaceutical agents on angiogenesis is commonly evaluated by results of in vivo assays, i.e. chick chorioallantoic membrane and rabbit cornea assays. However, reported results are not always objective. So, the aim of our study was to elaborate methods of estimation of endothelial cell proliferation as one of important stages of angiogenesis. PMID- 16706144 TI - [The influence of mesenchymal stem cell transplantation time on myocardial reparation in rat experimental heart failure]. AB - Mesenchymal stem cells (MSCs) have an ability to migrate in the organism to injured tissue to exert influence on inflammation and reparation in these regions. The aim of this study was to determine the optimal time of MSCs transplantation for myocardial reparation in rat experimental heart failure. The experiments were carried out on inbred line Wistar-Kyoto rats. Myocardial experimental infarction (EI) was induced by ligation of the left descending coronary artery. MSCs were isolated from bone marrow, cultivated in vitro and injected into the tail vein on the day of experimental infarction operation. It was shown that the time of MSCs transplantation exerted an essential influence on angiogenesis in a damaged myocardium, on ventricular dilatation and morphological structure of the scar. The best time for MSCs transplantation was determined within two days before EI, and seven days after EI. As a result, the overload of the border zone of infarct region decreased, and no features of infarction relapse were shown in the border zone. PMID- 16706145 TI - [Ultrastructure of the testes and spermatogenesis in the mite Anystis baccarum]. AB - The epithelial lining of testes in Anystis baccarum is glandular and produces a secretory product necessary to form spermatophores. The main stages of spermatogenesis occur in the lumen of the testis in groups of synchronously developing sister cells. Spermatogonia and late spermatids are encircled by glandular cells. Reorganization of developing spermatids is typical of the trombidiform mites and includes formation of the acrosomal complex, cytoplasm elimination, disappearance of the nuclear envelope and formation of invaginations of plasmalemma. The chromatin material condensation is not followed by the entire chromatin body formation. In mature spermatoza, dense chromatin strands (80b nm in diameter) lie along the cell in the peripheral layer of the cytoplasm. Mature spermatozoa lack axonema or any protrusions. A layer of microtubules, visible underneath the outer membrane, may serve for sperm movement in the female genital duct. The acrosomal complex consists of acromal granule, acrosomal filament and subacrosomal substance. This, as well as two aggregates of typical mitochondria, looks plesiomorphic. PMID- 16706146 TI - [The fine structure of chromatin in Paranosema grylli (Microsporida)]. AB - Nucleosomes were found for the first time in the nuclear chromatin of Microsporida--organisms known among the smallest eukaryotes on Earth. Chromatin of Paranosema grylli sporoplasm was studied by Miller's technique. On low ionic strength cell spreads, this chromatin was represented by 10 nm nucleosome filaments, 20 nm filaments, and "smooth" (nucleosome-free) filaments of 3-4 nm in diameter. Nucleosome filaments display structural heterogeneity seen as irregular arrangement of nucleosome particles along the filament length. Different nucleosome filaments show 13-30 nucleosomes per 1 microm with the length of linker DNA ranging from 10 to 45 nm. The present results suggest that microsporidian chromatin is weakly condensed. Only lower-order chromatin packaging levels displayed some structural peculiarities. PMID- 16706147 TI - [Differential gene expression in the jellyfish Aurelia aurita]. AB - The body of Aurelia aurita, as well as other diploblasts, consists of two epithelial layers: ectodermal and gastral epithelium. These two tissues are separated by mesoglea, or extracellular matrix. In most coelenterates mesoglea is acellular. In A. aurita mesogleal cells are scattered in mesoglea. Differential display PCR was used to compare mRNA pools from ectodermal epithelium, gastral epithelium and mesoglea. 4 novel gene fragments were cloned and sequenced. According to RTPCR results, one of these fragments is differentially expressed in the ectodermal epithelium. PMID- 16706148 TI - [Phosphorylation of proteasomes and alpha-RNP from rat liver cells]. AB - In eukaryotic cells the population of proteasomes is heterogeneous. Here we have shown that proteasomes from nuclei and cytoplasm of rat liver cells differ in their subunit patterns. The subunit pattern of alpha-RNP differs from that of proteasomes, however, alpha-RNP particles contain the number of 26S proteasome subunits. Moreover, the proteasomes contain subunits of alpha-RNP. We have shown for the first time that nuclear proteasomes and alpha-RNP are hyperphosphorylated on threonine residues. Differences in phosphorylation state of subunits of nuclear and cytoplasmic proteasomes and alpha-RNP on threonine and tyrosine residues have been revealed. A suggestion is put forward that hyperphosphorylation of subunits may determine nuclear localization of these complexes in rat liver cells. The results obtained suggest that a highly specialized system of protein kinases and phosphatases may be involved in the regulation of phosphorylation state of different populations of proteasomes and alpha-RNP in rat liver cells. PMID- 16706149 TI - Inductive characteristics of proteins secreted by retinal cells. AB - The studies of the development of eye rudiments and formation of adult eye tissues have always been among priorities in developmental biology and then in developmental genetics, which is associated with the peculiarities of the development and structure of the eye. In the late 80s, it was established by the group of developmental factors of the Institute of Gene Biology of RAS that many differentiated tissues are able to produce proteins causing homologous differentiations in polypotent cells of early gastrula ectoderm. The aim of our present study was isolation of proteins secreted by mammalian and fish retinal cells and determination of their inductive properties in early gastrula ectoderm of Xenopus laevis. The sets of proteins secreted by retina induce tissues homologous to the inducer, that is, neural tissue, brain, retina, pigmented epithelium, and also lenses and ear vesicles. The retinal inductive proteins retain their homologous inductive capacity after lyophilization. Biological testing shows that a total mixture of proteins secreted by retinal cells induces in polypotent gastrula ectoderm of X. laevis a narrower spectrum of tissues than the fractions obtained from this mixture. The above-outlined results obtained in thecourse of investigations of inductive peculiarities of retina and its fractions help in the elucidation of questions concerning embryonic induction and factors determining it, as well as questions concerning the maintenance of tissue specifity and regenerative capacity of the tissue studied. PMID- 16706150 TI - [Changes in cytokine content in the peripheral blood of volunteers after their exposure to polychromatic visible and infrared light]. AB - Anti-inflammatory, immunomodulating and wound-healing effects of visible and infrared (IR) radiation from laser and non-laser sources are widely used in current medicine. However, the role of pro- and anti-inflammatory cytokines in development of these effects has been poorly studied. A randomized, placebo controlled, double blind study was made. Using ELISA, the content of 10 cytokines was studied in the peripheral blood of volunteers after a single and four daily irradiations of the sacral area (D = 15 cm) with polychromatic visible + IR polarized light (480-3400 nm, 12 J/cm2). The phototherapeutic sessions were accompanied by four blood exfusions for the study (to a total volume of 80 ml). In the control (placebo) group, irradiation was imitated, and blood samples of the same volume were drawn at the same time intervals as in volunteers of the main group. A fast decrease in the level of pro-inflammatory cytokines was revealed as soon as in 0.5 h after the irradiation. This level was retained until the end of the phototherapeutic course. At the parameters exceeding the norm, the contents of TNF-alpha, IL-6 and IFN-gamma fell, on average, by 34, 12 and 1.5 times, respectively. By the end of the course, the levels of IFN-gamma and of IL 12 decreased by 5 and 15 times, respectively. A fast decrease (by two-fold) was also characteristic of normal values of IL-6. Neither IL-1beta, nor IL-2 were detected in blood plasma of the examined people both before and after the irradiation. In parallel with a decrease in the proinflammatory factor levels the amount of anti-inflammatory cytokines was found to rise: that of IL-10--by 2.7 3.5 times in 0.5 h and at later terms at the initially normal parameters, and that of TGF-beta1--by 1.4-1.5 times at the initially decreased level. The IL-4 content did not change. A characteristic feature of the light effect was a fast rise of IFN-gamma amount--by 3.3-4.0 times in individuals with its initially normal level, with no changes in IFN-alpha content. The above-reported regularities of the light effects were also recorded at a direct (in vitro) irradiation of the examined volunteers' blood, as well as on addition of irradiated blood to a 10-fold volume of non-irradiated autologous blood, i.e. at a modeling of mixing, of a small amount of transcutaneously photomodified blood with its main circulating volume in the vascular bed of an irradiated person. Such a similarity of effects in blood following its irradiation in vivo and in vitro enables us to associate the fast changes of the cytokine content in the entire volume of peripheral blood with the transcutaneous photomodification of its small amounts, and with a "transfer" of the light effects by photomodified blood to the whole pool of circulating blood. PMID- 16706151 TI - [Intracellular and intercellular signal transduction pathways and age alterations of proliferative activity and apoptosis intensity in cells of immune system]. AB - The paper considers age-associated alterations of intracellular and intercellular cascades of transduction of proliferative, differentiating, pro- and antiapoptotic signals, their interaction and influence on proliferative activity, differentiation and apoptosis of the immune system cells. One of initial causes of these alterations is accumulation with age of a growing number of antigens exposed on the surface of antigen-presenting cells. As a result of chronic antigenic stimulation, caused by this factor, an insufficient quantity or a slowed down appearance of growth factor receptors (in particular, IL-2 receptor) and costimulation molecules, primarily CD28, on T-cells membrane is observed. Because of this proliferative and antiapoptotic signals, received by T-cells, have a smaller intensity that predetermine reduction of their proliferative activity, and also activity of telomerase, and a greater susceptibility to apoptosis. Permanent activation of immune system is also reflected in age-related increase of expression of CD95 and type I tumour necrosis factor receptor by lymphocytes (that aggravates their susceptibility to apoptosis), and in intensification of proinflammatory cytokine synthesis. The second main cause of alterations in the immune system is an age-related decrease in the synthesis of growth factors that are necessary for cell survival and proliferation. In particular, because of the lack of IL-7, apoptosis intensity of maturing T-cells increases in thymus. Thymic stromal cells remain without contact signals and growth factors generated by lymphocytes, and also undergo apoptosis that causes further reduction of T-lymphopoiesis. Similar events also occur in bone marrow that predetermines age-related decrease in B-lymphopoiesis and in telomerase activity of haemopoietic stem cells, and also their proliferative potential reduction. PMID- 16706152 TI - [cAMP-PKA signal pathway in the lower eukaryotes]. AB - The review summarizes current data regarding transduction mechanisms by cAMP-PKA signal pathway in unicellular eukaryotes. Conceptions of evolutionary origin of eukaryotic signal transduction systems are developed. PMID- 16706153 TI - [Changes in the expression of receptors of steroid hormones in the development of partial androgen deficiency of aging men (PADAM)]. AB - This work is devoted to the vital topic of the influence of partial androgen deficiency of aging men (PADAM) on the development of cells with androgen receptors. The results obtained in this study suggest a conclusion that the production of testosterone by some tumors and tissues of the peritumorous zone, which is accompanied by increased proliferative activity and disturbance of the regulation of the cell cycle, is caused by PADAM. The given changes are directed at compensating for testicular deficiency (in particular at overcoming the androgen-dependent stage of development of androgen-sensitive cells). These changes are a partial manifestation of metabolic syndrome (X-syndrome). The atypical cells, which unavoidably develop during metabolic syndrome, are dealt with by means of the immune system, whose capabilities become less and less adequate in the given circumstances. PMID- 16706154 TI - ["Stationary phase aging" of cell culture: an attempt of evaluation of growth medium "age" effect]. AB - Cell proliferation rate and 3H-thymidine labeling index of "young" (i. e. harvested in 3 days after subcultivation) cultured Chinese hamster cells (B11 dii FAF28 line) have been determined in growth medium conditioned by the same cells for various periods of time during their growth and subsequent "stationary phase aging" (medium of different "age"). Cells were serially cultured in Eagle's medium with 10 % bovine serum. The experiment was conducted as follows. The "young" cells were seeded in Carrel's flasks (4500 cells/cm2) with fresh growth medium and placed at 37 degreesC. At definite time intervals, media from 3 randomly selected flasks were filtrated and stored in small glass flasks at 4 degreesC. The cells from all 3 flasks were collected by trypsin treatment and counted with hemocytometer. During the period of 26 day cultivation we collected a set of media of different "age" corresponding to certain points of the growth and "stationary phase aging" curve of the culture. Then, the "young" cells in fresh medium were seeded into tissue culture plates with cover slips placed into wells of the plates (26,600 cells/cm2) and grown at 37degreesC, 5 % CO2 for 2 h. At this point, the medium was replaced with media of different "age". 22 h later (i. e. on the first day after seeding) cell density was evaluated microscopically in all the wells. On the next day (i. e. in 2 days after seeding) 3H-thymidine was added to every well to final concentration 1.85 x 10(4) Bq/ml. After next 24 h (i. e. in 3 days after seeding) cell density was counted again, and the medium was removed. The cover slips were rinsed with Hank's solution and air-dried. Autoradiography was performed in standard manner by photoemulsion exposing for 5 days and subsequent developing in amidol developer. The relative number of nuclei with 10 and more "grains" was revealed microscopically. Based on the obtained results, two basic parameters were evaluated for every "age" medium: 1) cell proliferation activity index calculated as log2 (N3/N1), where N1 - cell density on the first day after seeding, and N3 - the same parameter on the third day after seeding; 2) cell labeling index calculated as percentage of cells with nuclei labeled by 3H-thymidine during incubation from 2nd to 3rd day of cultivation. These two indexes for cell growth in different "age" media appeared to be highly correlating (R = 0.85). Besides, it was found that the observed "age related" diminishing of ability of the growth media of different "age" to stimulate proliferation of "young" cells cannot completely explain the "stationary phase aging" phenomenon (in particular, even for the "oldest" medium cell labeling index was 65 %). We conclude that the phenomenon is based on exactly intrinsic changes of cells, most likely on molecular level, though environmental effects cannot be entirely excluded. The authors are grateful to the Russian Basic Research Foundation for support (grants 03-04-49030 and 00-04 48049). PMID- 16706155 TI - [Extrinsic cells, immunoreactive to Ca-binding protein, as sources of thalamic visual centres in tortoises]. AB - Extrinsic sources of calcium-binding proteins involved in immunoreactive innervation of the visual thalamic nuclei Rot and GLd in turtles (Testudo horsfieldi and Emys orbicularis) were studied using HRP tracing method and immunohistochemistry. In 1.5-4.5 months after monocular enucleation calbindin (Calb)-, parvalbumin (Parv)- and calretinin (Calr)-labeling was found in fragments of degenerated retinal fibers in the contralateral optic tract and in some retinorecipient structures (optic tectum, GLd and GLv). Changes in GLd were detected in its neuropil part. in 2.0-3.5 months after unilateral ablation of tectum and pretectum, the densities of Parv-, Calb- and Aclr-immunoreactivity terminals and fibers were diminisched in the ipsilateral n. Rot, with the maximum effect seen in Parv. Following HRP injection into the visual thalamus (Rot and GLd), retrogradely labeled cells with Parv label only, were revealed in the ventrothalamic nucleus Enta, pretectal nucleus Ptv, and in all types of Ca binding proteins (CaBPr) in separately labeled cells of the optic tectum. Thus, it has been shown that thalamic visual centers in turtles have multiple extrinsic cells, which serve as sources of CaBPr projections. The present data suggest that organization of CaBPr inputs to visual thalamus in reptiles (turtle) and higher amniotes are fundamentally similar. PMID- 16706156 TI - [Karyotype and morphology of larvae of Chironomus (Chironomidae, Diptera) from the Caspian Sea]. AB - Karyotype and morphology of Chironomus sp. larvae from the Caspian Sea have been described. 2n = 8. Chromosomal arm combination is AB, CD, EF, G (cytocomplex thummi). All chromosomes display conjugation of homologues. Centromere areas are of s-type. The nucleolar organizer and two Balbiani rings are disposed in arm C, and another Balbiani ring is in arm B. Half larvae have heterozygotic paracentric inversions in arm D. The larva belongs to salinarius form. One scleritis is colorless, an occipital scleritis is light and irregularly colored. Premandible has 4-5 cogs. Epipharingeal ridge has 18-23 dens. This new species is close to salinarius group, especially to Ch. albidus Konst., but differs from it at the larva stage in the structure of non-massive basal antenna segment, in the presence of large quantity of premandible denses, in light color of occipital scleritis, and in a series of morphometric rates. A similarity berween Ch. paraalbidus and Ch. albidus, on the one hand, and Boeotendipes, on the other one, has been found out. PMID- 16706157 TI - [Kinetic parameters of single ion channels and stationary conductivities of phytotoxin modified lipid bilayers]. AB - As shown earlier, phytotoxins produced by Pseudomonas syringae pv. syringae form ion channels of "small" and "large" conductance when incorporated into planar lipid membranes. The multilevel conductance is due to cluster organization of the channels (Kaulin et al., 1998; Gurnev et al., 2002). In this study the kinetic parameters of syringomycin E (SRE) and syringostatin A (SSA) channels in negatively charged bilayer lipid membranes were estimated. The average time of open state of the small channels (t(s)(open)) did not depend on transmembrane voltage (in the range of +/- 200 mV). The channel characteristics differed between two phytotoxins: the t(s)(open) for the SRE-channels was much larger than that for SSA-channels. An energetic diagram with two non-conducting states illustrating the formation of the small channel is proposed to explain the voltage independence of the kinetic parameters. The probability for synchronous functioning of small channels with SSA was higher than that with SRE. To analyse the role of the clusters in the biological activities of SRE and SSA, we estimated the cluster contribution to a net transmembrane currents to be 60 and 90%, respectively. PMID- 16706158 TI - [Effect of estradiol on stimulated theophylline and prolactin Ca2+ exit from intracellular stores of pig oocytes]. AB - Effect of estradiol on stimulated theophylline and prolactin Ca2+ exit from intracellular stores of pig oocytes was investigated using fluorescent dye chlortetracycline. It was shown that in the presence of estradiol neithert theophylline nor prolactin stimulated Ca2+ exit from intracellular stores of oocytes. Unlike, the common action oftheophylline and prolactin, also in the presence of estradiol, stimulated Ca2+ exit from intracellular stores. Inhibition of protein kinase C inhibits Ca2+ exit from intracellular stores in common action of theophylline and prolactin. These data suggest an obvious influence of estradiol on Ca2+ exit from intracellular stores of pig oocytes stimulated by theophylline and prolactin. PMID- 16706159 TI - [Coherent oscillations of the molecular state of protein in live cells]. AB - The paper describes results of experimental studies of changes in the refractive index of the live cell (human erythrocyte) substance. The measurements were carried out using automated interference microscope. Oscillations of the refractive index were detected with the amplitude 5-10% and period 1 x 10(3)-3 x 10(3). In other words, oscillations with a near one hour period is equal to 10-45 min. It is supposed that these oscillations may result from coherent changes in the state of protein molecules. PMID- 16706160 TI - [Alkaline phosphatase in Amoeba proteus]. AB - In free-living Amoeba proteus (strain B), 3 phosphatase were found after disc electrophoresis of 10 microg of protein in PAGE and using 1-naphthyl phosphate as a substrate a pH 9.0. These phosphatases differed in their electrophoretic mobilities - "slow" (1-3 bands), "middle" (one band) and "fast" (one band). In addition to 1-naphthyl phosphate, "slow" phosphatases were able to hydrolyse 2 naphthyl phosphate and p-nitrophenyl phosphate. They were slightly activated by Mg2+, completely inhibited by 3 chelators (EDTA, EGTA and 1,10-phenanthroline), L cysteine, sodium dodecyl sulfate and Fe2+, Zn2+ and Mn2+ (50 mM), considerably inactivated by orthovanadate, molybdate, phosphatase inhibitor cocktail 1, p nitrophenyl phosphate, Na2HPO4, DL-dithiothreitol and urea and partly inhibited by H2O2, DL-phenylalanine, 2-mercaptoethanol, phosphatase inhibitor cocktail 2 and Ca2+. Imidazole, L-(+)-tartrate, okadaic acid, NaF and sulfhydryl reagents -p (hydroxy-mercuri)benzoate and N-ethylmaleimide - had no influence on the activity of "slow" phosphatases. "Middle" and "fast" phosphatases, in contrast to "slow" ones, were not inactivated by 3 chelators. The "middle" phosphatase differed from the "fast" one by smaller resistance to urea, Ca2+, Mn2+, phosphates and H2O2 and greater resistance to dithiothreitol and L-(+)-tartrate. In addition, the "fast" phosphatase was inhibited by L-cysteine but the "middle" one was activated by it. Of 5 tested ions (Mg2+, Cu2+, Mn2+, Ca2+ and Zn2+), only Zn2+ reactivated "slow" phosphatases after their inactivation by EDTA treatment. The reactivation of apoenzyme was only partial (about 35 %). Thus, among phosphatases found in amoebae at pH 9.0, only "slow" ones are Zn-metalloenzymes and may be considered as alkaline phosphatases (EC 3.1.3.1). It still remains uncertain, to which particular phosphatase class "middle" and "fast" phosphatases (pH 9.0) may belong. PMID- 16706161 TI - [Karyotype and karyofund of chironomids Polypedilum (Pentapedilum) sordens (Diptera, Chironomidae)]. AB - Karyotypes and karyofund of chironomids P. Polypedilum (Pentapedilum) sordens (Van der Wulp, 1874) from four water bodies of Saratov region have been analysed. 2n = 6, I (GAB) > II (CD) > III (EF). A cytophotochart of polytene chromosomes of this species was composed, and discovered inversions were identified on its base. The level of inversion polymorphism is high. There are 18 chromosome sequences in the karyofund. The frequency of heterozygote inversions is 1.6-2.5 per specimen. On the base of this cytochart it is possible to produce chromosome charts of other species of subgenus Pentapedilum. PMID- 16706162 TI - [Hsp70 chaperone and the prospects of its application in anticancer therapy]. AB - Major stress protein Hsp70 is known to possess two important properties: ATP dependent activity and protective activity; these two are thought to play a significant role in anticancer therapy. Many malignant tumors contain high amounts of intracellular Hsp70. Moreover, many anticancer drugs themselves are able to elevate Hsp70 expression in tumor cells. Since Hsp70 was found to disturb many signal pathways of apoptosis in many points, the high chaperone expression may lead to an increased resistance of tumor cells to anticancer drugs. On the other hand, when overexpressed by a certain mechanism, Hsp70 is able to emerge at the cell surface by itself or together with tumor antigens and present these to immune cells T-lymphocytes and natural killers, in such a manner that makes cancer cell recognized and abolished. These properties make Hsp70 very promising instrument in designing some novel anticancer vaccines. PMID- 16706163 TI - [Human fetal ventricular cardiomyocytes in vitro: proliferation and differentiation]. AB - In this study, in the primary cell culture of human fetal cardiomyocytes proliferation of myocytes combines with their differentiation. The cells were isolated enzymatically from 19-22 week-old human fetuses and cultured for 14 days. DNA synthesis, ultrastructure and presence of atrial natriuretic peptide (ANP) were examined. In 7 day-old culture, the myocytes make about 60%, in 14 day old culture--about 50%. Myocytes synthesize DNA and divide mitotically. After a 24 h incubation with 3H-thymidine in 7 day-old culture 1.8 +/- 0.5% of muscle and 25.2 +/- 11.7% of non-muscle cells are labeled, in 14 day-old culture--2.5 +/- 0.5 and 8.1 +/- 1.7% of cells are labeled, respectively. In 7 and 14 day-old cultures the degree of redifferentiation of contractile apparatus in myocytes varies from scattered actin and myosin filaments surrounded by ribosomes to differentiating myofibrils with distinct sarcomeres and Z-discs. Single electron dense granules, morphologically similar to secretory atrial granules, display ANP immunoreactivity. Thus, human fetal ventricular cardiomyocytes in cell culture proliferate, differentiate and synthesize ANP for 14 days; this is indicative of vitality of these cells. PMID- 16706164 TI - [Characteristics of human embryonic neuronal cells procured by non-enzyme method]. AB - Isolation and culturing of human neuronal progenitor cells is of significant value for both fundamental research and therapeutic purposes. In this work, human embryonic neuronal cells were characterized as a heterogeneous population of progenitor cells with various differentiation potentials. During in vitro culturing the cells are capable of re-inoculating, proliferating, differentiating and migrating. While differentiating, these cells form neurons and glial cells. The present research demonstrates that depending upon the culturing conditions the embryonic neuronal cells may either form floating aggregates (incubation with embryonic serum), attach (incubation without serum), proliferate, or form neurospheres. Besides, peculiarities of aggregate differentiation during their incubation under various media are described. PMID- 16706165 TI - [Protective role of autotypic contacts under cerebellar neural net injury by toxic doses of NO-generative compounds]. AB - In the present work, cerebellar neural net injury was induced by toxic doses of NO-generative compound (NaNO2). A protective role of glial cells was revealed in such conditions. The present results were compared with those of the previous work concerning the action of high concentration glutamate on the frog cerebellum (Samosudova et al., 1996). In both cases we observed the appearance of spiral like structures--"wrappers)"--involving several rows of transformed glial processes with smaller width and bridges connecting the inner sides of row (autotypic contact). A statistic analysis was made according to both previous and present data. We calculated the number and width of rows, and intervals between bridges depending on experimental conditions. As the injury increased (stimulation in the NO-presence), the row number in "wrappers" also increased, while the row width and intervals between bridges decreased. The presence of autotypic contacts in glial "wrappers" enables us to suppose the involvement of adhesive proteins--cadherins in its formation. The obtained data suggested that the formation of spiral structures--"wrappers" may be regarded as a compensative adaptive reaction on the injury of cerebellar neural net glutamate and NO generative compounds. PMID- 16706166 TI - [The balance between Hsp70 and its cochaperones Hdj1 and Bag1 determines its substrate-binding activity]. AB - Heat shock protein Hsp70 presents one of the most effective cell protective systems. Its protective activity is mostly due to the fact that Hsp70 is able to restore native conformation of newly synthesized or damaged proteins. Two other proteins. Hdj and Bag 1, are involved in the process, allowing Hsp70 to perform binding-release cyclec of target proteins. The aim of this study was to investigate interactions between cochaperones Hdj 1 and Bag 1, and the major cell chaperone Hsp in vitro. The accumulation of Hsp70 and Hdj 1 in human erythroleukemia K562 cells was stimulated by heat stress (43 degrees C, 60 min). Cells were collected at certain time periods after heat stress, and amounts of cell chaperones were measured using Western blotting and ELISA assay. The level of Hsp70 chaperone activity in cell extracts was estimated using original technique. The effects of exogenous cochaperones and of their parts on this activity were also investigated. The results of the study indicate that Hsp70 chaperone activity is regulated by the level of its cochaperones, especially Hdj 1. At the same time the amount of ATP appears to be critical for functional activity of Hsp70. Hdj 1 and Bag 1 peptides, which bind to Hsp70 with high affinity, are able to significally reduce its chaperone activity. This finding confirms the possibility of using peptide approach for regulation of Hsp70 function at the cellular and organismal levels. PMID- 16706167 TI - [Respiration of wheat root cells under simultaneous inhibition of parts I and III of the electron transport chain of mitochondria by rotenone and antimycine A]. AB - Respiration of excised roots of 5 day old wheat seedlings with blocked mitochondrial oxidation under simultaneous action of rotenone and antimycine A was studied. A reduced rate of oxygen uptake was observed within the first hour of root treatment inhibitors. However, after a 5 h exposure there was an increase in oxygen uptake, which was prevented by KCN but amplified by malate and ascorbate. The application of inhibitors caused a considerable increase in the respiratory coefficient (RC) up to 2.1, that suggests a significant CO2 release, when the initial sites of mitochondrial electron transport chain were inhibited. RC did not raise, when ascorbate was added in the presence of inhibitors. We assume that inhibition of mitochondrial oxidation at I and III sites of electron transport chain facilitates switching on the alternative paths of reductant translocation to oxygen. Participation of ATPases and redox system of plasma membrane in the response reactions of respiration directed to the restoration of ion, particularly, proton homeostasis in conditions of inhibited mitochondrial oxidation is discussed. PMID- 16706168 TI - [Localization of NADPH-diaphorase in the brain of the shore crab Hemigrapsus sanguineus]. AB - The presence and localization of NADPH-diaphorase in the cerebral ganglion of the shore crab Hemigrapsus sanguineus was investigated with histochemical and electron histochemical methods. The reactivity of this enzyme was found in the deutrocerebrum, mainly in neuropils of olfactory lobes, the lateral antennular neuropil, a laterodorsal group of cells, and in the oculomotor nerve nucleus. Ultrastructural localization of the enzyme was detected in neurons on the perinuclear membrane, and in membranes of endoplasmic reticulum, in mitochondria and cytosol. The enzyme was found in axons of the antennular nerve, and in terminals of receptor axons in the glomerulus. The obtained data testify to participation of NO in perception and processing of the olfactory information. PMID- 16706169 TI - [Specific DNA sequences are associated with nuclear envelopes of pseudonurse cells in Drosophila melanogaster otu 11]. AB - Nuclei of ovarian pseudonurse cells from the mutant strain of Drosophila melanogaster otu 11 are suitable for mapping the attachment of chromosomes to the nuclear envelope (NE). Loci in contact with the NE included region 20CD of the X chromosome, region 41 of chromosome 2, the proximal end of region 81 of chromosome 3, and region 101 of chromosome 4. In situ hybridization revealed that all 4 regions contained sequences homologous to clone lambda20p1.4. DNA of clone lambda20p1.4 was previously found to bind specifically to purified D. melanogaster lamins. These results suggest that specific DNA sequences are involved in attachment of chromosomes to NE in vivo. PMID- 16706170 TI - [A combined approach to mapping the proximal border of the right arm of polytene chromosome 2 in Drosophila melanogaster otu 11]. AB - A combined approach based on cytological observations in situ hybridization, and qualitative Southern-blot analyses were used to localize the proximal border of the right arm of polytene chromosome 2 in Drosophila melanogaster otu 11 strain. A genetically functional chromosome 2 is bounded by "deletions" C', C, D, B, A and ms2-10. Using in situ hybridization in conjunction with comparative quantitative Southern-blot hybridization to deletions in centromeric heterochromatin, DNA of specific centromeric clone lambda20p1.4 was localized with respect to "deletions" and on otu 11 polytene chromosomes. Comparison of hybridization sites of lambda20p1.4 on polytene chromosomes, and its amount in mutant lines of D. melanogaster carrying known "deletions" in the centromeric heterochromatin enabled us to localize the proximal border of the right arm of chromosome 2 in D. melanogaster otu 11 strain between the 39/40 region and hybridization site of the k20p1.4 DNA fragment. PMID- 16706171 TI - [Karyotype structure in species of Propsilocerus genus (Diptera, Chironomidae)]. AB - Karyotype structure and polytene chromosome banding patterns were studied in two Orthocladiinae siblings--Propsilocerus akamusi (China) and Propsilocerus jacuticus (Russia). Both species have haploid number of chromosome typical for Orthocladiinae (n = 3). An unusual structure of centromeric regions was observed in all three chromosomes of karyotypes in both species. Photomaps of polytene chromosomes are presented. A comparison of karyotypes of P. akamusi and Propsilocerus jacuticus revealed a high level of homology in their banding sequences, however, the presence of fixed paracentric inversions in chromosomal arms IR, IIR, IIIR of Propsilocerus jacuticus has shown a clear-cut phylogenetic divergence. No chromosomal polymorphism was found in both species. PMID- 16706172 TI - [The relationship between nuclear-cytoplasmic transport and chromosome segregation]. AB - Chromosomes/chromatids transport to cell division poles (chromosome segregation) and nuclear-cytoplasmic transport give way to each other during cell cycle. Disassembly and reassembly of the nuclear envelope during input or output mitosis are the critical time periods for changing one kind of transport system for the other one. The interest in the problem of relationships between nuclear cytoplasmic transport and chromosome segregation came into view after revealing some factors involved in both transport systems. Among these factors are GTPase Ran, nuclear transport receptors and associated molecules, kinetochore proteins and proteins of nuclear pore complexes (nucleoprin, for example), spindle microtubules, microtubule motor proteins, and signal molecules. The role of all of these factors in the transport systems is not known. A decision of this problem will make it possible to determine mechanisms of both transport systems and to understand the character of evolutionary relationships between these systems. PMID- 16706173 TI - [The origin of eukaryotic cells and origination of apoptosis]. AB - The unified conception of the origin of eukaryotic cells has been proposed. In the author's opinion, evolutionary transformation of prokaryotic cell into eukaryotic cell took place 3.3-1.4 billion years ago and involved the next four stages: 1) the appearance of intracellular membranes due to prokaryotic cell plasmalemma invaginating into its cytoplasm; 2) the cell nucleus formation by the double sheet of intracellular membrane surrounding and sequestrating genetic material of the cell; 3) the appearance of cytoskeleton in parallel with mitotic spindle formation and gradual transition from prokaryotic way of cell division to mitosis; 4) the establishment of symbiosis between the evolving nucleated cell and prokaryotic microorganicsms that subsequently transform into mitochondria and chloroplasts. Apoptosis of cells of the present day multicellular eukaryotic organisms is supposed to be an evolutionary altered response of mitochondrian predecessors to the influence of factors, which are able to damage eukaryotic host cell. The initial biological significance of this reaction pertained to attempts of endosymbionts to leave the host cell as soon as possible, if the probability of its irreversible injury was very high, and by this to escape from their death. It is possible that numerous proteins, known as sensors or transducers of proapoptotic signals in Bcl-2--p53-dependent apoptotic pathway, were initially encoded by mitochondrial genome, whereas antiapoptotic factors and also components of receptor-mediated and granzyme B perforin dependent apoptotic pathways have cellular origin. PMID- 16706174 TI - [Continuous human embryonic stem cell lines]. AB - A new continuous human embryonic stem cell line (HESC-5) derived from a blastocyst is described. The cultured cell passed over 200 population doublings, which exceeds the Hayflick's limit sufficiently. The cells maintained a stable proliferative activity, high activity of alkaline phosphatase, and expression of transcription factor Oct-4 and of surface antigens SSEA-3, SSEA-4 and TRA-1-60 known to be characteristic of embryonic stem cells of the human origin. Immunofluorescent detection of antigens, characteristic of ectoderm, endoderm and mesoderm in the new cell line HESC-5, and in the previously described other four stem cell lines confirms the ability of these cells to retain their pluripotency under in vitro condition. In addition, in all the cell lines, a high telomerase activity was revealed, which controls a stable telomere length and, hence, an unlimited ESC proliferation. Unlike other cell lines, HESC-5 was found, under specific conditions, to spontaneously differentiate into hematopoietic cells. A morphological similarity was shown between ESC colonies cultivated both on a feeder layer and in the non-feeder system. PMID- 16706175 TI - [Characteristics of human mesenchymal stem cells isolated from bone marrow and adipose tissue]. AB - Populations of human mesenchymal stem cells were derived from bone marrow and adipose tissue. Here analysis of six individuals is represented. Cells were isolated, expanded and evaluated by the expression of surface antigens using flow cytometry. These cells displayed similar characteristics for many markers. Cells isolated from bone marrow and adipose tissue were found to be homogeneously positive for CD13, CD44, CD90, CD105, and negative for CD45, CD34, CD31 and CD117. Besides, differences in surface antigene CD10 expression between narrow and adipose tissue-derived cells were detected. All these findings indicate that both bone marrow and adipose tissue are important sources of mesenchymal stem cells, which could be used in cell therapy protocols. PMID- 16706176 TI - [Typical changes of lymphocyte surface architectonics in chronic viral infection]. AB - With the use of scanning electron microscopy, lymphocyte surface architectonics has been described in cases of chronic viral persistence at tick-borne (vernal) encephalitis, hepatitis types A and B, and Herpes simplex. In lymphocytic population smooth and plicate cells in addition to cells with vesicles and outgrowths on the surface prevailed. The appearance of lymphocytes with a combined type of superficial microrelief was registered. It is supposed that despite the variety of types of lymphocyte surface architectonics and possible causes promoting their polymorphism, changes in superficial ultrastructure of immunocompetent cells for chronic viral infection have a nonspecific character. PMID- 16706177 TI - [Phenomena of "micronucleoli" in cardiomyocytes from patients with idiopathic restrictive cardiomyopathy]. AB - Silver staining was used to estimate structural and functional conditions of cardiomyocyte and stromal cell nucleoli in myocardium from 3 patients with idiopathic restrictive cardiomyopathy (RCM). We revealed a prominent increase in the amount of myocardial stromal component cells. The area of nuclei, the area and amount of nucleoli in cells of myocardial stromal component much varied. In some cells of myocardial stromal component we found nuclei, whose area was comparable with that of cardiomyocyte nuclei. In all patients we revealed cardiomyocytes with extranucleolar distribution of argentophilic substance in the nuclei. These data suggest an important pathogenic role of structural rearrangments in the nuclei of cardiomyocytes and stromal component cells in providing specific cell phenotype of myocardium in patients with idiopathic RCM. Extranucleolar (atypical) distribution of argentophilic substance in the nucleus, referred to as "micronucleoli", point out certain disturbances of nucleolar functions in cardiomyocytes in patients with RCM. PMID- 16706178 TI - [Revelation and identification of laminin in the structure of plasma membrane of ascitic Zajdela hepatoma cells]. AB - Cell dysdifferentiation during neoplastic transformation is a crucial problem of cell biology and oncology. Antigenic diversion of cancer cells is a typical characteristic of dysdifferentiation. It involves the appearance of antigens which are unusual for normal tissue of this type. Components organospecific for membrane proteins of normal kidney were previously found among plasma membrane proteins of hepatocellular rat tumors, rat hepatocytes after carcinogen treatment, and regenerating liver, respectively. In the present work we showed that a protein with mol. weight about 200 kDa reacting with laminin-1 immunoserum is the basic component of plasma membranes of the rat Zajdela hepatoma cells, which is responsive for organospecific anti-kidney immunoserum in Western blot. A mass-spectrometer analysis of trypsin proteolysis fragments was carried out in SDS-PAGE slices containing the investigated component. The analysis showed the presence of beta1, beta2 and alpha4 laminin chains peptides. The component with mol. weight about 180 kDa, found in the Western blot with laminin-1 immunoserum, was also subjected to the mass spectrometer analysis. As a result, a gamma1 laminin chain was found. An increased amount of laminin was revealed in the ascitic liquid and sera of rat with developed Zajdela hepatoma, in comparison with sera of normal rats. In addition, we found the appearance of laminin on the hepatocyte surface on the 4th day after hepatocarcinogen injection (N diethylnitrosamine, DENA). Thus, for the first time tumor associated antigens were revealed and identified in the structure of plasma membranes of Zajdela hepatoma cells, being specific to rat kidneys. Our results allow to conclude that in the process of carcinogenesis in rat liver laminin synthesis occurs, which is also characteristic of the rat hepatoma Zajdela cells. PMID- 16706179 TI - [Lectin and mitotic activities in root meristems of winter wheat under oryzalin effect]. AB - The effect of oryzalin, a microtubule polymerization inhibitor (10 MM), on lectin and mitotic activities (mitotic index and duration of mitotic phases) was studied in unhardened (23 degrees C) and hardened (7 days, 2-3 degrees C) winter wheat seedlings. Three wheat cultivars differing in their frost tolerance were compared. Oryzalin treatment (3 h) decreased activity of soluble lectins, increased activity of cell wall lectin mitotic index. Under these conditions, prolongation of anaphases and disappearance of telophases were detected. Plant hardening reduced the sensitivity of cell wall lectins and mitotic activity to the cytoskeleton inhibitor due, presumably, to the appearance of cold-stable microtubules. Plant growing and hardening with oryzalin stopped mitoses and caused the appearance of polyploid cells and cells with micronuclei. These abnormalities were preserved after hardening. The results obtained demonstrate an important role of microtubules in adaptation of plants to low temperature. PMID- 16706180 TI - [Effect of testosterone concentration on the intensity of 5alpha dihydrotestosterone and 17beta-oestradiol formation]. AB - The culture of human male foreskin fibroblasts was incubated with various concentrations of testosterone in the medium. It has been shown that the minimum formation of 17beta-oestradiol and 5alpha-dihydrotestosterone occurs at physiological testosterone concentrations. Any deviation of testosterone concentration, both up and down, was accompanied by an increase in the formation of 17beta-oestradiol and 5alpha-dihydrotestosterone. PMID- 16706181 TI - [SRE, NF-kappaB, and AP-1 DNA-binding activities induced by A431 cell adhesion correlate with actin cytoskeleton reorganization]. AB - Cell adhesion to extracellular matrix proteins induces activation of different signal molecules and influences gene expression. As shown earlier, epidermoid carcinoma A431 cell adhesion to fibronectin, laminin-2/4 or antibodies to receptor EGF (ab EGFR) results in reorganization of specific cell shape and actin cytoskeleton in the majority of cells. This study resolves a question whether morphological changes are accompanied with some cell response at the level of gene expression in nuclei. We have shown that cell reattachment promotes a specific DNA-binding of nuclear extracts with consensus sequences SRE, NF-kappaB and AP-1, compared to the control. NF-kappaB and AP-1 activities were considerably reduced in spread cells, which did not show actin filament's structures typical for the ligands. SRE specific proteins demonstrated other peculiarities and depended on the type of immobilized ligands. Our results argue that actin cytoskeleton reorganization, induced by cell adhesion to immobilized ligands at the early period after cell reattachment, is correlated with a specific answer at the levels of DNA-binding activity of transcription factors SRE, NF-kappaB and AP-1. PMID- 16706182 TI - A forum for supply chain technology. PMID- 16706183 TI - Prevent sepsis from taking its toll. Interview by Robert Neil. PMID- 16706184 TI - Finding the wrong fit. PMID- 16706185 TI - Pandemic preparedness. Vendors and hospitals alike face challenges as well as unknown factors. PMID- 16706186 TI - Hospitals equip to meet the bariatric challenge. Rising number of obese patients necessitates specific supplies. PMID- 16706187 TI - SWOT team solves supply chain issues. PMID- 16706188 TI - A supply delivery system that can streamline your case cart operation. PMID- 16706189 TI - [The influence of nitric oxide inductor (NaNO2) on the sensitivity of Chinese hamster cells to hyperthermia]. AB - Formerly we reported the reduction of sensitivity to gamma- and UV-radiation in Chinese hamster cells of line 90 under the influence of nitric oxide inductor (NaNO2). Of interest was to learn if it possible to reduce the influence of hyperthermia on cells by means of NO inductor. A 1 h long incubation with this NO donor demonstrated an increased survival of cells heated up to 45 degrees C, and a decreased frequency of chromosomal aberrations in these. Employment of cycloheximide (CHE), an inhibitor of protein synthesis, and administration of CHE together with nitric oxide donor (NaNO2), equally increased the cell survival at hyperthermia. These and relevant literature data suggest that the demonstrated effect of exogenous NO may be associated with HSP70 protein. The noticed decrease in the number of chromosomal aberrations in heated cells under the influence of NO donor may play an important role in its modifying effect on cells at hyperthermia. PMID- 16706190 TI - [Seasonal rhythmics of the leucoplast structure in bulb scales of Scilla sibirica L]. AB - A study was made of seasonal changes in plastids of ground tissue cells of bulb scales in early-spring ephemeroid Scilla sibirica L. In summer, plastids are represented by typical amyloplasts, with their main volume (97.0 +/- 4.3%) being occupied by one large starch grain. The volume fraction of plastid stroma is at its minimum. The stroma contains small plastoglobuli and no thylakoids. The same structure is characteristic of plastids in October. However, no starch is found in December, when some thylakoids are seen at the plastid periphery. In the early spring (March), when leaves still remain below the ground, the volume fraction of starch grains is 53.0 +/- 2.2%. In the stroma some structures superficially similar to those of microtubuli are revealed. The thylakoid system is fairly well developed, some of thylakoids being concentrically arranged. Some electron-opaque material is seen in the thylakoid lumen. Many plastids are sheathed with elements of the smooth endoplasmic reticulum. Based on the analysis of these and literature data, a conclusion is made that plastids of bulb scales not only store starch, but also seemingly participate in phytohormone biosynthesis. PMID- 16706191 TI - [Interaction of laminin with the plasma membrane components of ascitic Zajdela hepatoma cells]. AB - The laminin affinity chromatography was used for isolating laminin-binding proteins from the plasma membrane of Zajdela hepatoma cells synthesizing laminin. These were components with mol. weights about 80, 67, 60, 55, 52, 48 and 43 kDa. The isolation of laminin integrin receptors from plasma membranes of Zajdela hepatoma cells in the presence of MnCl2 detected only a protein with mol. weight about 80 kDa in EDTA-elution conditions. This protein was identified by mass spectrometry method as the 78 kDa glucose-regulated protein precursor (GRP78). It belongs to the family of 70 kDa heat shock proteins, recently GRP78 was reported to be localized on the surface of different cell types, including hepatocytes. PMID- 16706192 TI - [Detection and identification of a tumor-associated heteroorganic antigen of Zajdela hepatoma in non-histone proteins of chromatin]. AB - By polyacrylamide gel electrophoresis, a phosphoprotein with mol. weight of 42 kDa was detected in non-histone proteins (NHP) of chromatin of Zajdela ascitic hepatoma cells eluted from phosphocellulose with 0.4-0.5 M NaCl. A protein of the same mol. weight is present in narrow fractions of rat kidney chromatin, but is absent in rat liver. It is suggested that the revealed protein corresponds to the tumor-associated heteroorganic NHP antigen detected earlier in NHP chromatin of rat tumor cells. By MALDI mass spectrometry, this phosphoprotein was identified as ERK2/mitogen-activated protein kinase. PMID- 16706193 TI - [Induction of premature senescence program by an inhibitor of histone deacetylase sodium butyrate in normal and transformed rat fibroblasts]. AB - We investigated a possibility to induce the premature cell senescence in rat embryo fibroblasts and E1A + cHa-ras transformants. We found that after the treatment with sodium butyrate, an inhibitor of histone deacetylases, both normal and transformed cells completely stopped to proliferate and accumulated at G1/S and G2/M phases of the cell cycle. The cloning efficiency data show that the cell cycle arrest induced by sodium butyrate is irreversible and correlates with the accumulation of active phosphorylated form of stress kinase p38, and with the expression of marker of senescence--beta-galactosidase activity (SA beta-Gal). The program resembling the premature senescence after sodium butyrate treatment is supposed to develop both in normal and transformed cells. The irreversible block of proliferation in E1A + cHa-ras transformants may be regarded as an example of activation of anticancer program like that of premature senescence in the tumor rodent cells. PMID- 16706194 TI - [E1A oncogene effect on the ability of p21(Waf1) to regulate G1/S arrest in E1A expressing transformants following irradiation]. AB - P21(Waf1) cyclin-dependent kinase inhibitor blocks cell cycle transition from G1 phase into DNA replication after DNA damage. The main targets of p21(Waf1) are Cyc 1E--Cdk2 and Cyc 1A--Cdk2 complexes, PCNA (proliferating cell nuclear antigen), a subunit of DNA polymerase delta, and E2F-1 transcription factor. The universal mechanism of cell cycle arrest in normal cells is determined as p21(Waf1) interaction with positive regulators of G1 phase. As a rule, DNA integrity control mechanisms are destroyed in the process of oncogenic transformation, which results in proliferation of genetically defective cells. The purpose of our study was to investigate molecular mechanisms of cell cycle regulation in transformants that are able (E1A + E1B-19kDa) or unable (E1A(+) + cHa-ras) to be arrested at G1/S checkpoint. We have shown that p21(Waf1) is able to form complexes with cyclins and Cdks, PCNA and E2F-1 transcryption factor, although it interacts with E1A oncoproducts in both transformants. The presence of E1A bound p21(Waf1) in cyclin-kinase complexes seems to be the cause of activating phosphorilation of Cdk2 at Thr-160 in cyclin A/E--Cdk2 complexes in both control and X-ray irradiated cells. Thus, the absence of G1/S arrest following irradiation in E1A + cHa-ras transformants and its presence in E1A(+) + E1B-19kDa transformants is not connected with differences in interaction of p21(waf1) with the main regulators of G1-to-S transition, but is realized through other not yet identified ways. PMID- 16706195 TI - [The role of different kinase pathways of signal transduction in proliferation of E1A + Ras transformants]. AB - In this paper we have explored the role of different kinase pathways of signal transduction in proliferation control of E1A + Ras transformants, using specific inhibitors of MAP-kinases ERK, JNK, p38 and PI3-kinase. According to our data, suppression of signalling cascades driven by RI3K only arrested proliferation of E1A + Ras cells, while suppression of either MAP-kinase did not lead to noticeable antiproliferative effect. We have shown that suppression of RI3K with LY294002 gave rise to accumulation of cyclin-dependent kinase inhibitor p27(KiP1) but not p21(Waf1). Accumulation of p27(KiP1) in LY294002-treated E1A + Ras cells was accompanied by a decrease in Cyclin E-Cdk2 and Cyclin A-Cdk2 activity, which caused diminution of Rb phosphorylation and strengthening of E2F-Rb binding. Binding of E2F with hypophosphorylated Rb resulted in inhibition of E2F activity and reduction of E2F-regulated gene transcription, these genes being necessary for S-phase entry and DNA synthesis. Thus, RI3K--Akt cascade plays the key role in maintenance of autonomous proliferation of cells transformed with E1A and cHa ras oncogenes. Inhibition of PI3K leads to p27(Kip1) accumulation and cell cycle arrest, consequently. PMID- 16706196 TI - [Subunits of human holoenzyme of DNA dependent RNA polymerase III phosphorylated in vivo]. AB - Phosphorylation of human holoenzyme of DNA dependent RNA polymerase III subunits in vivo has been investigated. RNA polymerase III from human placenta nuclei and epidermoid carcinoma cells A431 was isolated as two subfractions (IIIa and IIIb) distinguished in the order of elution from DEAE Sephadex A-25 column chromatography and buoyant density at glycerol gradient centrifugation. The subfractions of RNA polymerase III holoenzyme consists of four subunits with molecular masses 60, 52, 45 and 38 kDa, respectively, phosphorylated in vivo. The subunits with molecular masses 60 and 45 kDa are phosphorylated both on tyrosine and serine/threonine residues. All these subunits belong to subunits of the molecules of RNA polymerase III proper. RNA polymerase III and RNA polymerase I have the 38 kDa subunit in common. The subunit with molecular mass 52 kDa is phosphorylated on serine/threonine residues and may be related to some basal transcription factors of RNA polymerase III. PMID- 16706197 TI - [Energy metabolism in wheat root cells under modification of plasma membrane permeability by antibiotic nystatin]. AB - This paper reports changes in ion transport and energy metabolism of plant cells during short- and long-term expositions, resp., to antibiotic nystatin, which is known to specifically bind with plasma membrane sterols to form channels. The excised roots of 5 days old wheat seedlings were used as a model system in this research. It has been shown that treatment of excised roots with nystatin leads to activation of energy metabolism expressed as an increase of respiration and heat production by root cells. Furthermore, in the presence of nystatin increased pH of incubation medium, plasma membrane depolarization and a significant loss of potassium ions were observed. Nystatin-induced stimulation of respiration was prevented by malonate, an inhibitor of succinate dehydrogenase, electron acceptor dichlorophenolindophenol, and AgNO3, an inhibitor of H(+)-ATPase. Based on the data obtained it can be suggested that nystatin-induced stimulation of respiration is related to electron transport activation via mitochondrial respiratory chain, and is connected with activation of plasmalemma proton pump. Moreover, nystatin-induced increase of oxygen consumption was prevented by cerulenin, an inhibitor of fatty acid and sterol synthesis. This indicates that additional sterols and phospholipids may be synthesized in root cells to "heal" nystatin-caused damage of plasma membrane. A supposed chain of events of cell response to nystatin action may by as following: formation of nystatin channels influx of protons--depolarization of plasmalemma-efflux of potassium ions disturbance of ion homeostasis--activation of H(+)-ATPase work-increase in energy "requests" for H(+)-ATPase function--increase in the rate of oxygen consumption and heat production. The increased energy production under the action of nystatin, may provide the work of proton pump and synthesis of sterols and phospholipids, which are necessary for membrane regeneration. PMID- 16706198 TI - [Structural dynamics, stability and folding of proteins]. AB - The present concepts of protein folding in vitro are reviewed. According to these concepts, amino acid sequence of protein, which has appeared a result of evolutionary selection, determines the native structure of protein, the pathway of protein folding, and the existence of free energy barrier between native and denatured states of protein. The latter means that protein macromolecule can exist in either native or denatured state. And all macromolecules in the native state are identical but for structural fluctuations due to Brownian motion of their atoms. Identity of all molecules in native state is of primary importance for their correct functioning. The dependence of protein stability, which is measured as the difference between free energy of protein in native and denatured states, on temperature and denaturant concentration is discussed. The modern approaches characterizing transition state and nucleation are regarded. The role of intermediate and misfolded states in amorphous aggregate and amyloid fibril formation is discussed. PMID- 16706199 TI - [Physical-chemical properties of actin in different structural states. New ideas about its folding-unfolding pathways]. AB - Results of actin folding-unfolding pathways examination and characterization of intermediate and misfolded states are summarized. Properties of microenvironments and peculiarities of location of tryptophan residues in protein are analysed in detail. This allowed to conclude that the main contribution to the bulk fluorescence of native protein is made by internal tryptophan residues Trp 340 and Trp 356, localized in hydrophobic regions, while tryptophan residues Trp 79 and Trp 86 are quenched. It has been shown that inactivated actin, previously regarded as an intermediate state between native and completely unfolded state of protein is in reality a misfolded aggregated state. The properties of actin in this state were characterized in detail. In particular, it is shown that inactivated actin is a monodisperse associate consisting of 15 monomer unit. Two earlier unknown intermediate states, which precede completely unfolding of protein macromolecule and formation of inactivated actin, were visualized. A new scheme of folding-unfolding processes was proposed. It is shown that the reason of anomalous effects, which are recorded for actin in solutions with small concentrations of GdnHCl, is a specific interaction of actin with a denaturant. PMID- 16706200 TI - [Room temperature phosphorescence of amorphous aggregates and amyloid fibrils resulting from protein misfolding]. AB - Using actin, alpha-lactalbumin and insulin as examples, it was shown that the formation of amorphous aggregates of proteins and amyloid fibrils leads to an increase in the rigidity of tryprophan and tyrosine residues micro-environment and, consequently, to the appearance of tryptophan (tyrosine) room temperature phosphorescence (RTP). RTP was used for examining a slow intramolecular mobility of native (G-, F-form) and inactivated (I) rabbit skeletal muscle actin during the process of GdnHCl induced protein unfolding. This method made it possible to confirm that an essentially unfolded intermediate precedes the formation of inactivated actin. It has been found that the kinetic intermediate generated at the early stage of protein denaturation has no tryptophan RTP, suggesting a high lability of its structure. Symbate changes of integral intensity (relative quantum yield) and the mean lifetime of RTP during the U*-->I transition suggest a gradual increase of the number of monomers incorporated in the associate (U*- >11...-->In...-->I15), which is accompanied by an increase of protein structural rigidity. The rate of inactivated actin formation (I-->I15) is shown to increase with the increase of protein concentration. It is shown that, no matter what method of inactivation was employed (1--2 M GdnHCl or 3.0-3.5 M urea, Ca2+ removal, incubation at 70 degrees C, refolding from completely unfolded state by dialysis from 8 M urea or 6 M GdnHCl), actin transition to the inactivated state is accompanied by a significant increase in both integral intensity and the mean lifetime of RTP, suggesting the rigid structure of inactivated actin. It is shown that the lifetime of inactivated actin RTP does not depend on GdnHCl concentration within the limits from 0 to 4 M. On using insulin and alpha lactalbumin as examples, it is shown that RTP can be used in studies of fibrillogenesis and properties of amyloid fibrils. PMID- 16706201 TI - [The structure and stability of the glutamine-binding protein from Escherichia coli and its complex with glutamine]. AB - A study was made of the conformational changes in the Escherichia coli glutamine binding potein (GlnBP) induced by GdnHCl, and of the effect of glutamine (Gln) binding on these processes. Intrinsic fluorescence, ANS emission fluorescence, and far- and near-UV circular dichroism spectroscopy were used. The obtained experimental data were interpreted, taking into the account results of the analysis of tryptophan and tyrosine residues microenvironments. This enabled us to explain the negligible contribution of Tyr residues to the bulk fluorescence of the native protein, the similarity of fluorescence characteristics of GlnBP and GlnBP/Gln, and an uncommon effect of the excess of fluorescence intensity at 365 nm (Trp emission) upon excitation at 297 nm compared to the excitation at 280 nm. The latter effect is explained by the spectral dependence of Trp 32 and Trp 220 contributions to protein absorption. The dependence of Trp fluorescence of protein on the excitation wavelength must be taken into account for the evaluation of Tyr residues contribution to the bulk fluorescence of protein, and in principle, it may also be used for the development of an approach to decomposition of multi-component protein fluorescence spectrum. The parametric presentation of fluorescence data showed that both GlnBP unfolding and GlnBP/Gln unfolding are three-step processes (N-->I1-->I2-->U), though in the case of the GlnBP/Gln complex these stages essentially overlap. Despite its complex character, GlnBP unfolding is completely reversible. In comparison with GlnBP, in the case of GlnBP/Gln the dramatic shift of N-->I1 process to higher GdHCl concentrations is shown. PMID- 16706202 TI - [Conformational changes of disulfide isomerase C induced by guanidine hydrochloride]. AB - Unfolding--refolding of Escherichia coli disulfide isomerase C (DsbC) induced by GdnHCl was studied by intrinsic fluorescence. Interpretation of experimental fluorescence data was done together with the analysis of protein 3D structure. It is shown that although Cys 141 is the next neighbour of a single tryptophan residue Trp 140, sulfur atoms of the disulfide bond Cys 141--Cys 163 are far apart from the indole ring and cannot quench its fluorescence, while the potential quenchers are Met 136 and His 170. It has been revealed that, though each subunit of DsbC contains eight tyrosine residues, only three tyrosine residues (Tyr 171, Tyr 38 and Tyr 52) contribute to the bulk fluorescence of the molecule. The character of intrinsic fluorescence intensity changes induced by GdnHCl (equilibrium and kinetic data), the character of parametric dependencies between fluorescence intensity recorded at 320 and 365 nm, and the existence of an isosbestic point of protein fluorescence spectra in solutions with different GdnHCl concentrations, allowed suggesting a one-step character of DsbC denaturation. The reversibility of this process is also shown. PMID- 16706203 TI - [The role of quarternary structure in fluorescent protein stability]. AB - The stability of fluorescent proteins (FPs) is of great importance for their use as reporters in studies of gene expression, protein dynamics and localization in cell. A comparative analysis of conformational stability of fluorescent proteins, having different association state was done. The list of studied proteins includes EGFP (monomer of green fluorescent protein, GFP), zFP506 (tetramer GFP), mRFP1 and "dimer2" (monomer and dimmer of red fluorescent protein), DsRed1 (red tetramer). The character of fluorescence intensity changes induced by guanidine hydrochloride (GdnHCl) of these proteins differs significantly. Green tetramer zFP506 has been shown to be more stable than green monomer EGFP, red dimmer "dimer2" has been shown to be less stable than red tetramer DsRed1, while red monomer mRFP1 has been shown to be practically as stable as tetramer DsRedl. It is concluded that the quaternary structure, being an important stabilizing factor, does not represent the only circumstance dictating the dramatic variations between fluorescent proteins in their conformational stability. PMID- 16706204 TI - [Participation of transfused bone marrow cells in reparative osteohistogenesis]. AB - The participation of skeletal tissue cell precursors in the repairing regeneration of bone tissue was studied. Bone marrow was taken from donor animals -mice of C57Bl/6-TgN(ACTbGFP) 1 Osb line (The Jackson Laboratory Bar Harbor ME USA line). Nucleated cell fraction was isolated by centrifugation on a density percoll gradient. Recipient mice C57Bl/6 line were irradiated by 7.0-7.5 Gr dose. Intravenous infusion of donor cells and osteoclasts of tibia was done after irradiation of recipient mice. Histological preparations of bone regenerate tissues were studied on 15, 30, and 60 days by confocal microscopy. Donor cells were found as skeletal tissue precursors into periost, endost, bone marrow, and as differentiated cells of newborn tissue of regenerate--osteoblasts, osteocytes, chondrocytes. The data obtained indicate that part of donor bone marrow cells are able to progressive differentiation under recipient bone fractures. PMID- 16706205 TI - [Dynamics of microtubular cytoskeleton in higher plant meiosis. IV. Middle prometaphase]. AB - By means of morphological analyses of meiotic abnormalities in pollen mother cells of cereal distant hybrids, processes of cytoskeleton cycle at the middle prometaphase (chaotic stage) were studied. It was shown that elements of the bipolar spindle (central and opposite kinetochore fibres) are formed at the chaotic stage of meiotic prometaphase. PMID- 16706206 TI - [The structural and functional analysis of the surface apparatus in four species of Sarcocystis (Sporozoa, Apicomplexa)]. AB - The comparable ultrastructural analysis of the sarcocyst surface apparatus (SSA) was made for four species of Sarcocystis: Sarcocystis muris, S. fusiformis, S. medusiformis, and Sarcocystis sp. from buffalo heart muscles. In all these species, SSA contains a surface membrane, overmembrane complex with glycocalyx, and submembrane complex made of two glycoprotein SSA primembrane layers. SSA makes numerous primary vesicle-like protrusions and pits in between. Some vesicles containing two layers, PM1 and PM2, are pinching off from the totally formed protrusions. Then these vesicles are directed into infected host cell to participate in its degradation. In the SSA pits neither over-, nor submembrane complex is present, the pits being made of the surface membrane only. It is important that fibrillar structures penetrate through the SSA membrane into pits from the host cell. Besides, SSA forms secondary protrusions with different structures in various species of Sarcocystis. They increase the sarcocyst surface and transport different substances along intermediate filaments from the SSA pits membrane to the sarcocyst body. At the same time, deep invaginations are found in the SSA of old sarcocysts. We thought that these structures increased the sarcocyst surface and thus promote to intensify metabolism. This study-defined presence of membranous vesicles in secondary protrusions. According to their structure and localization, the membranous vesicles may be involved in the building of the sarcocyst surface membrane. PMID- 16706207 TI - [Effect of EGF on the activity of nuclear and cytoplasmic 26S proteasomes in A431 cells]. AB - It has been first shown that EGF regulates a proteolytic activity of proteasomes. Following a 15 min action with 100 ng/ml EGF, three types of peptidase activity of both cytoplasmic and nuclear proteasomes were induced in A431 cells, although, this effect on different populations of proteasomes was selective. EGF preferentially stimulates chymotrypsin-like activity of cytoplasmic proteasomes, and induces a similar increase of chymotrypsin-like, trypsin-like and peptydylglutamyl peptide hydrolase activities of nuclear particles. Tyrphostin, an inhibitor of tyrosine kinase activity of EGF receptor, prevents the EGF effect on both proteolytic and RNase activity of nuclear and cytoplasmic proteasomes. It is concluded that EGF may rapidly and selectively stimulate enzymatic activity of EGF receptor. PMID- 16706208 TI - [Assembly of nucleolus-derived foci in various cultured mammalian cells during mitosis]. AB - During mitosis, rebuilding of the nucleolus is a step-wise process that, above all, includes an assembly of nucleolus-derived foci (NDF) in the cytoplasm of telophase cells. In this study, we performed a comparative analysis of NDF formation in mitotic cells of various mammalian cell cultures, such as green monkey CV1 cells, human HeLa cells, mouse 3T3 cells, and pig PK cells, both in control and following inhibition of rRNA synthesis by actinomycin D or by an adenosine analogue, DRB. The results obtained show that in all examined cell types NDF are formed shortly after or simultaneously with the onset of chromosome segregation to the poles of the mitotic spindle. However, an efficiency of NDF assembly, i.e. the number of NDF per anaphase or telophase cell, and the portion of anaphase and telophase cells containing NDF vary in different cell cultures, being most prominent in CVI and HeLa cells. In these cells, the vast majority of NDF accumulate several proteins of the mature nucleolus, such as B23/nucleophopmin, C23/nucleotin, fibrillarin, and, to a lesser extent, Nop52. The rRNA harbored by NDF is synthesized several hours prior mitosis, and plays an essential role the maintenance of NDF structural integrity. Starting from early stages of the assembly onwards, the NDF are predominantly located in the area occupied by aster microtubules of the mitotic spindle. PMID- 16706209 TI - [Structural organization and possible functional role of annulate lamellae containing cytoplasmic pores]. AB - This review is devoted to annulate lamellae, a specific compartment of endoplasmic reticulum that occurs, presumably, in actively growing and rapidly dividing cells (oocytes, embryonic and tumor cells). We summarized both earlier and recent data on the dustribution of annulate lamellae in various cell types, on their morphology, and the distribution of interaction with intracellular structures at various treatments. As the annulate lamellae contain cytoplasmic pore complexes, a special attention was paid to their relation with nuclear pores. Possible functions of the annulate lamellae in intracellular processes and, particularly, in nuclear envelope assembly, are discussed. PMID- 16706210 TI - ["Chain" karyotypic evolution of embryonic stem cell line R1 in vitro]. AB - Cytogenetic anomaly frequencies were analysed in three sublines of ES R1 line in its five clonal sublines, obtained from two cell colonies after transformation of ES R1 cells by plasmid with gene lif. Cell transformation did not increase cytogenic anomalies, however, the initial sublines of ES R1 line, as well as its transformed clonal descendants bore a redundant quantity of the chromosome 8 material within the structure of various Robertsonian translocations even in cells with diploid chromosome quantity (2n = 40). In the initial sublines ES R1 and its clonal descendants a common Rb (8; 15) was revealed. It was supposed that selection for the increase in ES cell sensitivity to cytokines (in particular, LIF) under cultural conditions was accompanied by an increase in chromosomal copies, carrying genes of mapk andjak/stat, through which downstream effectors of cytokine signals for preservation of cell pluripotention and propagation are realized. Genes of chromatid separation and chromosome segregation control (for example, separase gene Esp1 in chromosome 15) may be passively involved in this process, thus promoting acceleration of karyotype evolution in ES cells. PMID- 16706211 TI - [3-D reconstruction of early preimplantation mouse embryo]. AB - Method of 3-D reconstruction approaches for early mouse embryo in preimplantation stages was modified. The developed technique is based on application of light microscopy of serial thin sections and well known soft operating. The designed method enabled us 1) to get serial sections of a single mouse embryo; 2) to create an orthogonal system independent on the sample for orientation of virtual sections. The adequacy of 3-DR protocol was checked on reconstruction of air bubbles embedded in epoxy resin as a model of sphere. PMID- 16706212 TI - [Cytoskeleton abnormalities and meiotic restitution in the higher plants]. AB - Cytoskeleton abnormalities leading to the formation of restitution nuclei, and to the reunion of segregated daughter chromosome groups in the higher plant meiosis are reviewed. PMID- 16706213 TI - [Diffusional attenuation of proton echo and control of changes in the cytoskeleton hydration level]. AB - Using the pulsed NMR-method with pulse gradient of the magnetic field, a study was made of changes in diffusional attenuation (DA) of proton echo in germs of triticum grains under the influence of agents depolymerizing cytoskeleton elements or inhibiting their ATP-activity. At a short diffusion time, we observed respective alterations of NMR-population in a fraction of cell water molecules contributing to DA at large values of pulse gradient. It is shown that the presence in DA of a site with essentially smaller coefficient of self-diffusion, as compared with basic cell water fraction, is not a consequence of restricted diffusion phenomenon. The observed changes in NMR-population of the appropriate fraction of cell water molecules are interpreted as changes in the level of cytoskeleton hydratation occurring under the influence of the used agents. A hypothesis is proposed for the functional role of similar changes in the level of hydratation of cytoskeleton elements located inside plamodesmata in the mechanism of plasmodesmatal water permeability regulation. PMID- 16706214 TI - [Dependence of Ca2+ release from intracellular stores on NADH and FAD levels in fertilized and unfertilized bovine oocytes]. AB - Relation between NADH and FAD concentrations and the quantity of calcium released from intracellular stores in fertilized and unfertilized bovine oocytes was investigated using luminescent analysis. Inhibition of Ca2+ exit from intracellular stores was detected in degenerative oocytes at metaphase II and 2 cell embryos. The intensity of both NADH and FAD fluorescence increased in 2-cell degenerated embryos, whereas the increase in only NADH fluorescence intensity occurred in degenerated oocytes at metaphase II stage. Degeneration exerted no influence on NADH fluorescence intensity or Ca2+ exit from intracellular stores, whereas a decreased FAD fluorescence intensity was noted in degenerated pronuclei. The obtained data testify that in degenerated zygotes and early embryos Ca2+ release may occur from different intracellular stores. PMID- 16706215 TI - [Peculiarities of theophylline and prolactin interaction and calcium fluctuation from intracellular stores of porcine oocytes in the presence of estradiol]. AB - Using a fluorescent dye chlortetracycline, a study was made of the effect of estradiol on the interaction of theophylline and prolactin in the course of Ca2+ exit from intracellular stores of pig oocytes, isolated from ovaries at the stage of follicle growth. It is shown that in the presence of estradiol, prolactin does not stimulate Ca2+ exit from intracellular stores of pig oocytes. The action of theophylline similarly does not stimulate Ca2+ exit. Unlike, a joint effect of theophylline and prolactin on pig oocytes in the presence estradiol stimulated Ca2+ exit from intracellular stores of pig oocytes. These data demonstrated the influence of estradiol on theophylline and prolactin stimulated Ca2+ exit from intracellular stores of pig oocytes. PMID- 16706216 TI - [Inhibitory action of polycationic peptides on hormonal regulation of adenylyl cyclase of the ciliate Dileptus anser]. AB - To analyse molecular mechanisms of regulatory action of different hormones on the activity of the adenylyl cyclase signaling system (ACS) of the ciliate Dileptus anser, we studied the influence on this process of six synthetic polycationic peptides and peptides, corresponding to C-terminal regions of mammalian G-protein 385-394 alphas- and 346-355 alphai2-subunits. As we reported earlier, these peptides block hormonal signal transduction in tissues of the higher eukaryotes. Now it has been found that both polycationic peptides, containing hydrophobic C to-radicals, and branched peptides decrease regulatory effects of peptide hormones (insulin, relaxin) and biogenic amines (serotonin, adrenaline) on adenylyl cyclase (AC) activity and GTP-binding. In regard to the following peptides Cys-epsilonAhx-Trp-Lys-Lys(C10)-Lys2-Lys(C10)-Lys3-Lys(C10)-Tyr-Lys Lys(C10)-Lys-Lys-amide and [(Gly-Arg-Gly-Asp-Ser-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg Arg-Arg-Pro- Pro-Gly)2-Lys-EAhx-Cys]2 (epsilonAhx - E-aminocaproyl, C10 - caprinoyl group) their dose-dependent inhibitory action is shown. In cell culture of D. anser with a lower basal AC activity, both hydrophobic and branched peptides stimulated AC and GTP-binding without hormones. The data give evidence that these peptides can activate ACS of ciliates in a receptor-independent manner. No influence of peptides 385-394 alphas and 346-355 alphai2 on hormonal signal transduction in D. anser was observed, due, presumably, to some structural differences of G-proteins of the lower and higher eukaryotes. A conclusion was made about an important role of polycationic regions for functional coupling of hormone-activated receptor and G-proteins in the ciliate D. anser. PMID- 16706217 TI - [Peculiarities of metaphase chromosome methylation pattern in preimplantation human embryos]. AB - Methylation pattern peculiarities revealed by immunocytochemical analysis of metaphase chromosomes from preimplanted human embryos with monoclonal antibodies against 5-methylcytosine are described. Chromosomes of 2-8-cell triploid human embryos are undermethylated, if compared to those from PHA-stimulated fetal cord blood lymphocytes. Hemimethylation (asymmetric labeling of sister chromatids) is typical for the most of embryonic chromosomes at 2-cell--blastocyst stages due most probably to a passive loss of methylation during initial cleavages. Diffuse labeling and sister chromatid exchanges are two other cytogenetic peculiarities revealed by immunofluorescent staining of early human embryos. Hypomethylation of pericentromeric heterochromatin of chromosomes 1, 9, 16 and different methylation status of some homologous chromosomes may distinguish them from metaphase chromosomes of lymphocytes. M-banding pattern typical for chromosomes from adult and cord blood lymphocytes initially appears in embryonic metaphase chromosomes as early as at a 8-cell stage to be established for most part of chromosomes of the karyotype at the morula-blastocyst stage only. PMID- 16706218 TI - [Immunocytochemical analysis of human metaphase chromosome methylation status]. AB - The present paper describes a distribution of 5-methylcytosine-rich DNA in human metaphase chromosomes from PHA-stimulated lymphocytes. Immunocytochemical detection of 5-methylcytosine was carried out with monoclonal antibodies. Fluorescent signals were preferentially localized in certain chromosomal regions, corresponding to R-, some T-bands, pricentromeric heterochromatin, and short arms of acrocentric chromosomes. Specificity of fluorescent signals distribution along chromosomes allowed to describe a new type of human metaphase chromosomes banding pattern, which we call M-banding. Specific M-markers of landmarks were identified for each chromosome pair. The analysis of M-bands methylation status was carried out taking into account data available in literature on their nucleotide structure features, namely GC-rich H3 isochore content and CpG-islands concentration. According to our results, a high level of methylation is typical for the majority of GC-rich regions. However, certain bands of 6, 9, 10, 13, 15 chromosomes (6q15, 6q21, 6q23, 9p13, 9p22, 9p32, 10q24, 13q22, 15q15, 15q24) were shown to be hypomethylated, suggesting their special functional status in lymphocytes. PMID- 16706219 TI - [Differentiation of interphase domains of the homologous human X-chromosomes]. AB - In accordance with molecular biology data reported elsewhere, homologous interphase X-chromosome territories differ greatly in the abundance of inactive condensed chromatin. On the other hand, a three dimensional FISH (3D FISH) method has revealed that domains of both inactive and active X-chromosome have similar volumes and similar maximum section areas (Smax). To solve this contradiction, we examined differences between homologous human interphase X-chromosome territories using two dimensional FISH (2D FISH) preparations of clustered PHA-stimulated lymphocytes. For obtaining such preparations, we developed a new technique to avoid a stage of hypotonic treatment of living cells, since this treatment levels the chromatin compactness degree. According to our 2D FISH data, the mean ratios of Smax for larger and smaller homologous X-chromosomes, calculated for individual flattened nuclei, were 1.83 +/- 0.08 and 2.02 +/- 0.09, respectively, for clumped cells and groups of loosely associated and separated lymphocytes. In comparison, the same ratio calculated for individual 3D nuclei of PHA-stimulated lymphocytes was 1.38 +/- 0.05 (Falk et al., 2002). Our findings give evidence for enrichment of inactive X-chromosomes by low stretchable condensed chromatin. In addition, these findings show that an active X is enriched by a high stretchable form of chromatin, whose content may distinctly vary from cell to cell. PMID- 16706220 TI - [Karyological studies of two populations of Juniperus communis L. in west Siberia]. AB - Results of a karyological study of Juniperus communis L. populations under swamp and dry conditions are presented. The chromosome number of J. communis are 2n = 22. Analysis of morphological chromosome parameters showed a similarity between karyotypes of both populations. It is possible to identify one pair of asymmetric chromosomes (VIII pair); this chromosome pair is close to submetacentric type. Three pairs of chromosomes (I, VII, VIII) have secondary constrictions. Other metacentric chromosomes form groups of five long (II--VI) and three short (IX-XI) pairs. Differences between two populations in absolute chromosomal length are observed. PMID- 16706221 TI - [Organization of cytokinesis in the plant cell division]. AB - This review involves consideration of morphological and functional aspects of cytokinesis, such as role of microtubule and actin cytoskeleton as well as mechanisms of cell plate formation and phragmoplast expansion, spatial control of division plan and cell cycle regulation, in addition to newly discovered mutations. Recent progress in understanding mechanisms of cytokinesis in the plant cell is discussed. PMID- 16706222 TI - [The influence of plasma membrane ion permeability modulators on superoxide formation and bioelectrical characteristics of wheat root cells]. AB - Changes in superoxide radical formation and bioelectrical characteristics of excised wheat root cells under modification of plasma membrane ion permeability were studied. It was shown that a 2 h treatment of excised roots with valinomycin (Val, 20 microM), N, N'-dicyclohexylcarbodimide (DCCD, 100 microM), gramicidin S (Gr, 20 microM), chlorpromazine (CPZ, 100 microM) caused an increased loss of potassium by cells, lowering of membrane potential (MP) and electrical input resistance (Rin) of the cells. The superoxide formation by excised root cells diminished (under DCCD) or remained at the control level (under Val), which was accompanied by a minor decrease of MP and Rin of the cells, a small increase in potassium loss by excised roots, and in no change of pH of incubation medium. Significant depolarization of plasma membrane, dropping of Rin and essential loss of potassium ions by the cells correlated with a rise in the medium alkalinization and superoxide formation by excised roots (in the presence of Gr, CPZ). Ion channel blocker gadolinium (Gd3+, 200 microM) caused an increase of MP and Rin reduction of potassium loss by cells, and a decrease of pH of the incubation medium, and also enhancement of superoxide formation by excised root cells. It is suggested that upon plasma membrane ion permeability modification the activity of superoxide generating systems depends on the specificity and mechanisms of action of modulators, and is determined by their influence on redox state of plasma membrane as well as by peculiarities of ion transport disturbance. PMID- 16706223 TI - [Abnormalities of the plant division spindle]. AB - A list of 24 abnormal plant spindles observed in meiotic division of PMCs in wide hybrids, meiotic mutants, haploids, polyploids has been presented. The abonormalities arranged according to the course of normal prometaphase are provided with descriptions of their morphogenesis and consequences, microphotographs and schemes. The spindle abnormalities leading to meiotic restitution are registered. PMID- 16706224 TI - [Tumor cell proliferative activity in aggressive human lymphomas: the influence of reactive cell admixture on the assessment of proliferative indices]. AB - Uncontrolled proliferation is one of the main features of tumor cells, and therefore proliferative indices provide prognostic information, which might be valuable for treatment selection. However, in aggressive human lymphoid tumors, an admixture of normal (reactive) cells may be available in all organs infiltrated by neoplastic cells. We have presented data on determining proliferative activity using Ki-67 immunostaining, and demonstrated that an admixture of reactive cells could exert significant influence on the assessments of proliferative indices. We developed an experimental approach, which makes it possible to select neoplastic cells from the overall lymphoid population and to assess their proliferative activity - tumor cell proliferative indices. Tumor cell proferative indices determined in large cell lymphomas might be significantly higher than proliferative indices of overall populations, and the use of tumor cell proliferative indices will result in a more accurate assessment of disease prognosis. PMID- 16706225 TI - [Changing extracellular matrix of cells during development of suspension culture of Triticum timopheevii Zhuk]. AB - A study was made of the contents of the main polysaccharide fractions in the cell wall, and extracellular polysaccharides, and of the activity of cell wall enzymes during cultivation of suspension culture of cells of the winter wheat Triticum timopheevii Zhuk. It was shown that within 3 days of cultivation (a phase enriched in dividing cells), on the background of increased callose contents in plant cells, amounts of pectins and hemicelluloses extracted by 4N alkali decreased. The content of polysaccharides reached its initial level by the end of culturing. A parallel analysis of glycosidase activity in cell walls has shown their considerable activation at the stage enriched by dividing cells, which decreased at a transition of culture into the stationary level. The increased activity of hydrolyzing enzymes was combined with an increased efflux of extracellular polysaccharides into culture medium. The detected changes in polysaccharide composition of the cell wall at the first phase indicate its qualitative changes during cell wall reconstruction at the beginning of cytokines, whereas extensive expansion of cell wall was seen on the phase of elongation. PMID- 16706226 TI - [Agglutination and phagocytosis of foreign abiotic particles by hemocytes of the blowely, Calliphora vicina in vivo. I. Dynamics of hemocyte activity during larval development]. AB - Three types of Calliphora larval hemocytes have been revealed to be involved in phagocytosis of abiotic foreign particles: thrombocytoids, larval plasmatocytes and plasmatocytes I. Thrombocytoids are the quickest to respond to the appearance of invaders. The onset of test particle entrapment by thrombocytoid cytoplasmic fragments was observed, depending on the larval age within 0.5-5.0 min after injection. Separated fragments were fused, forming strands or roundish agglutinates. Phagocytosis of carbon, carmine or Indian ink particles by larval plasmatocytes occurs far more lately, and no earlier than 20-30 min after injection. Plasmatocytes I are capable of foreign particles adhesion on their surface, with a subsequent morule formation, and of engulfing these particles. These two events start in different time periods: adhesion occurs in 5-10 min, while phagocytosis is observed in 1--3 h. The rate of test particle entrapment and stability of agglutinales clearly depends on the larval age. The most pronounced reaction of hemocytes to foreign particles may be observed by the end of feeding and crop emptying. The second, somewhat less expressed rise of activity occurs in mature larvae not long before the onset of pupariation. Diapause induction is accompanied by reducing activities of both plasmatocytes and thrompocytoids. The importance of different hemocyte types in cellular immune reaction of Calliphora vicina larvae, and co-ordination between plasmatocytes and thrombocytoids are discussed. PMID- 16706227 TI - [Genetic nature of abnormal larval development in the progeny of l(1)ts403(sbr10) females of Drosophila melanogaster]. AB - In Drosophila melanogaster the small bristles (sbr) gene is vital and evolutionary conservative and controls nuclear export of mRNA. Sbr mutant alleles had a broad pleiotropic effect. High frequency of abnormal larva dying (up to 18 %) at the first instar stage in progeny of heat shock (37 degrees C, 1 h) treated mutant females is one of the most interesting l(l)ts403(sbr10) allele effects. Abnormal larvae display characteristic phenotype that involves the Malpighian tubules defect. Using interphase FISH method (fluorescence in situ hybridization), we showed that abnormal larvae had monosomy on chromosomes 2 and 3. DNA content in neuroblast interphase nuclei of abnormal larvae is 2.1 times less than in normal larvae. We suggest that abnormal larvae could be full or mosaic haploids that appeared as a result of maternal genome loss during fertilization or the mitotic division. Larvae with the same abnormalities appear in a progeny of females with different genotypes mating with males carrying compound chromosomes 2 or 3. FISH analysis showed that such larvae had monosomy only on a chromosome that is compound in paternal strain. Thus, monosomy on large autosomes may cause aspecial phenotype of abnormal larvae in D. melanogaster. PMID- 16706228 TI - [Comparative analysis of cell populations with a phenotype similar to that of mesenchymal stem cells derived from subcutaneous fat]. AB - Two cell populations with a phenotype similar to that of mesenchymal stem cells (MSC) with different characteristics for expression of surface antigene CD34 were derived from subcutaneous fat. CD34-positive cells were derived from subcutaneous fat of the inferior eyelid obtained during transconjuctival blepharoplasty. CD34 negative cells were derived from adipose tissue obtained during lipoaspiration from the same patients. These cells displayed common characteristics for morphology and expression of basic markers characterizing them as mesenchymal stem cells. On being induced for differentiation, these two cell populations were able to differentiate to cells of adipose (adipocytes), bone (osteoblastes, osteocytes), cartilage (chondroblasts, chondrocytes), and nervous (neurons, astrocytes and oligodendrocytes) tissues. PMID- 16706229 TI - The capacity of mesenchymal stem cells for neural differentiation in vitro. AB - It has been shown that mesenchymal stem cells (MSCs) of bone marrow from newborn rabbits can be induced for neuronal differentiation. The epidermal growth factor (EGF) introduced in the culture at the rate of 2 ng/ml is able to promote differentiation of neurons from bone marrow mesenchymal stem cells in 27 days of cultivation. Differentiated cells were marked by monoclonal antibodies to 70 kDa neurofilaments. The data obtained show a possibility of using bone marrow stem cells in therapy of neurodegenerative diseases. PMID- 16706230 TI - [The role of proton pump and potassium channels of plasma membrane in regulation of cellulose synthesis in plants]. AB - A study was made of the influence of two activators of plasma membrane proton pump [indole-3-acetic acid (IAA) and fusicoccin (EC)] and of the blocker of potassium channels of outward direction [tetraethylammonium chloride (TEA)] on exogenous [U-14C]glucose incorporation into cellulose fraction of cell wall, and on the value of plasmalemma membrane potential. It has been shown that IAA and FC exert different influences on the intensity of [U-14C]glucose incorporation into cellulose fraction: IAA activates, while FC inhibits incorporation intensity. A conclusion is made that differences in affects of IAA and FC on the intensity of cellulose synthesis at the plasma membrane level may be due to the fact that the activating effect of IAA on plasma membrane proton pump involves activation of the inward direction potassium channels, whereas that of FC, on the contrary, is associated with their blocking. Under the action of TEA, the intensity of incorporation of radioactively labeled glucose was increased. Apparently, the role of plasma membrane in regulation of the intensity of cellulose synthesis may be associated with not only the activity of proton pump on plasma membrane, but also the functional condition of potassium channels of this membrane. PMID- 16706231 TI - [EGF-dependent signaling pathways are activated under heat stress in A431 carcinoma cells]. AB - EGF receptor transactivation and activation of EGF-dependent signaling pathways under heat shock conditions were studied. Heating A431 cells at 42 degrees C induced both EGF receptor tyrosine phosphorylation and appearance of phosphorylated forms of key components of its downstream signaling pathways - phospholipase Cgamma1 (PLCgamma1), transcription factor STAT3, and EPK1/2. It is suggested that EGF receptor is transactivated under heat shock in A431 cells. Pretreatment of heat-shocked cells with a specific inhibitor of EGF receptor tyrosine kinase tyrphostin AG1478 does not prevent EGF receptor and EPK 1/2 tyrosine phosphorylation. In contrast, tyrphostin AG1478 abrogates tyrosine phosphorylation of PLCgamma1 and STAT3. This suggested that the intrinsic EGF receptor tyrosine kinase is not involved in EGF receptor transactivation, but is sufficient for PLCgamma1 and STAT3 activation in stress conditions. The effect of a conditioned medium of heated cells was investigated to check whether autocrine mechanism is involved in EGF receptor transactivation. The conditioned medium of heated cells induced both tyrosine phosphorylation of EFG receptor and ERK 1/2. Simultaneously, neither PLCgamma1, not STAT3 phosphorylation were detected. Here, for the first time, we demonstrated the involvement of autocrine mechanism in EGF receptor transactivation under heat stress in A431 carcinoma cells, but additional intracellular events are essential for activation of EGF receptor downstream signaling pathways. PMID- 16706232 TI - Non-medical prescribing. AB - In the UK, the Medicines Act 1968 limited the legal right to prescribe medicines to doctors, dentists and veterinary surgeons. Nearly 40 years later, non-medical prescribing, the extension of prescribing responsibilities to nurses, pharmacists and other healthcare professionals, is a key development in the NHS. Within England, from May 2006, certain nurse and pharmacist prescribers can prescribe any licensed medicine (except most controlled drugs) within their sphere of competence. Here we discuss the historical context, the different types and the clinical implications of non-medical prescribing. PMID- 16706233 TI - Bevacizumab and cetuximab for colorectal cancer. AB - Every year in the UK, around 16,000 people die from colorectal cancer, the second commonest cause of death from cancer in the UK after lung cancer. Over half of all people with colorectal cancer eventually die of metastatic disease. While median survival has increased with optimal use of combination chemotherapy, only a small minority of patients are still alive 5 years after diagnosis of metastases. Bevacizumab (pronounced be-va-see-zoo-mab) (Avastin - Roche) and cetuximab (se-tuks-ee-mab) (Erbitux - Merck) are two new monoclonal antibodies licensed for treating patients with metastatic colorectal cancer. Here we assess their efficacy and safety. PMID- 16706234 TI - Two ounces of prevention. PMID- 16706235 TI - Contact dermatitis following sustained exposure to pecans (Carya illinoensis): a case report. AB - Type I hypersensitivity reactions following ingestion of peanuts and tree nuts are well characterized. Cutaneous hypersensitivity reactions are less well characterized, yet they remain the second most common reaction pattern to contact with or ingestion of such nuts. We present a case of a patient who experienced an acute vesicular cutaneous reaction after prolonged contact with pecans. This case illustrates the salient features of contact dermatitis and serves as a reminder that contact with allergenic foods can lead to hypersensitivity reactions. PMID- 16706236 TI - Aquatic antagonists: lionfish (Pterois volitans). PMID- 16706237 TI - What is your diagnosis? Focal epithelial hyperplasia (Heck disease). PMID- 16706238 TI - Circumscribed juvenile-onset pityriasis rubra pilaris with hypoparathyroidism and brachyonychia. AB - Circumscribed juvenile-onset pityriasis rubra pilaris (PRP) manifests as well defined erythematous scaly plaques with follicular keratosis mainly over the elbows and knees. There are several reports of the association of PRP with other conditions. We report a boy with scattered erythematosquamous skin lesions and follicular hyperkeratotic papules since he was 6 years old. Results of a skin biopsy were compatible with PRP. The patient also had hypoparathyroidism and brachyonychia. To our knowledge, this association has not been reported to date, though minor disturbances of calcium and vitamin D metabolism have been mentioned in some disorders of keratinization. We further discuss the epidemiologic, clinical, and pathologic features of PRP; review the conditions associated with brachyonychia; and give a brief discussion about the possible role of calcium metabolism in disorders of keratinization. PMID- 16706239 TI - Biopsy may help identify early pyoderma faciale (rosacea fulminans). AB - Pyoderma faciale is an uncommon acute presentation of rosacea. Edema, nodules, and draining sinuses may occur. Women in their early 20s are typically affected, and severe scarring may result in untreated cases. We report the case of a woman in whom a biopsy helped establish an early diagnosis so that decisive intervention could be initiated and scarring avoided. The histologic features of pyoderma faciale only rarely have been described and are illustrated in this case. PMID- 16706240 TI - Community-acquired methicillin-resistant Staphylococcus aureus skin infection presenting as a periumbilical folliculitis. AB - Community-acquired methicillin-resistant Staphylococcus aureus (CAMRSA) infection is a clinical problem of increasing global incidence. CAMRSA most commonly presents as abscess and cellulitis of the skin and soft tissue. However, the lesions of cutaneous CAMRSA infection are pleomorphic and may appear as erythematous pustules of superficial folliculitis. This report presents the cases of 2 patients with CAMRSA skin infection that presented as a superficial folliculitis. The distribution of CAMRSA-related, erythematous, folliculocentric pustules was periumbilical, in contrast to the lesional location of methicillin susceptible S. aureus (MSSA)-associated folliculitis, which typically appears on the axillae, bearded area, buttocks, and extremities. CAMRSA should be considered in the diagnosis of periumbilical folliculitis or superficial folliculitis arising in areas not typically affected by MSSA-related folliculitis, such as the chest, flanks, and scrotum. PMID- 16706241 TI - Henoch-Schonlein purpura in an adult Filipino man: a case report and literature review. AB - Henoch-Schonlein purpura (HSP) is a form of cutaneous small vessel vasculitis that can involve visceral organs and is associated with deposition of immunoglobulin A (IgA)-containing immune complexes. HSP may appear after a remote history of infection (often an upper respiratory tract infection) as a rash with palpable petechiae or purpura. In addition, a patient with HSP usually complains of arthralgia and abdominal pain. Renal involvement also is common. HSP may be confused with other systemic autoimmune diseases because they all can present with similar symptoms. Prognosis is good and recovery usually occurs without treatment. Although HSP predominately affects children, the condition also can be seen in adults. We present a case of adult-onset HSP in an otherwise healthy Filipino man and review the literature. PMID- 16706242 TI - Extramammary Paget's disease resistant to surgery and imiquimod monotherapy but responsive to imiquimod combination topical chemotherapy with 5-fluorouracil and retinoic acid: a case report. AB - Extramammary Paget's disease (EMPD) is an uncommon skin neoplasm that usually affects the elderly population and occurs in the genital, anorectal, or axillary areas. The recommended treatment of EMPD involves surgical excision, including Mohs micrographic surgery; however, surgery is associated with a high rate of recurrence. There have been reports of successful treatment of recurrence with monochemotherapy involving topical imiquimod 5% cream. We report a case of EMPD recurrence after surgery that was resistant to imiquimod monotherapy but that completely resolved after imiquimod was combined with topical 5-fluorouracil (5 FU) and retinoic acid. To our knowledge, this is the first reported case of imiquimod combination therapy with 5-FU and retinoic acid for the treatment of recurrent EMPD. PMID- 16706243 TI - Predictors of healthcare outcomes and costs related to medication use in patients with acne in the United States. AB - This study investigated the relationship among health status, costs linked with the treatment of acne in the United States, and other aspects related to medication use. The US Medical Expenditure Panel Survey (MEPS) database was analyzed for a cohort of people with acne. This cross-sectional study obtained costs, demographics, healthcare service utilization, and clinical patient variables from the MEPS database. The EuroQol Group's EQ-5D scores available in MEPS were used for health status information. Multivariate weighted analysis was performed for data for approximately 5 million patients (weighted sample size). Nearly 70% of the patients used some type of medication for acne. Acne-related medication accounted for approximately 36% of the total acne-related annual healthcare costs, with an average of 2 annual acne prescription refills per patient. Increased number of refills of acne-related medications was associated with an improvement in health status (P<.05). Increased physician office-based visits were the only predictors of higher acne-related annual healthcare costs (P<.01). Adherence to acne medications is an important component of better health status. Pharmacologic treatment of acne does not significantly add to acne related annual healthcare costs. PMID- 16706244 TI - Efficacy and safety of once-daily metronidazole 1% gel compared with twice-daily azelaic acid 15% gel in the treatment of rosacea. AB - Rosacea is an inflammatory dermatologic disorder characterized by the presence of facial erythema, visible blood vessels, papules, and pustules. The National Rosacea Society has established a classification system that identifies 4 distinct rosacea subtypes based on clinical presentation: erythematotelangiectatic, papulopustular, phymatous, and ocular. The goal of topical therapy for rosacea is to reduce inflammatory lesion counts; decrease intensity of erythema; and reduce symptoms such as stinging, burning, and pruritus. Metronidazole and azelaic acid are thought to reduce the inflammation associated with rosacea by inhibiting the production of reactive oxygen species produced by neutrophils. Both metronidazole 1% gel and azelaic acid 15% gel recently have been approved for the treatment of rosacea. The current study was conducted to compare the once-daily application of metronidazole 1% gel with twice-daily applications of azelaic acid 15% gel for the treatment of patients with moderate rosacea (N=160). Both treatments showed similar reductions in inflammatory lesion counts (77% for metronidazole 1% gel and 80% for azelaic acid 15% gel) and high success rates in both global severity (53.7% vs 56.4% for metronidazole 1% gel and azelaic acid 15% gel, respectively) and erythema (42.7% vs 42.3% for metronidazole 1% gel and azelaic acid 15% gel, respectively). On average, the efficacy (including reduction in erythema) of the once-daily application of metronidazole 1% gel and twice-daily applications of azelaic acid 15% gel were similar. PMID- 16706245 TI - A look at the safety of metronidazole 1% gel: cumulative irritation, contact sensitization, phototoxicity, and photoallergy potential. AB - Rosacea is a common, recurrent, inflammatory dermatologic disorder characterized by the presence of facial erythema, visible blood vessels, papules, and pustules. The condition may cause serious psychologic morbidity and may significantly affect quality of life. The first topical rosacea therapy approved by the US Food and Drug Administration was metronidazole for the treatment of inflammatory lesions and erythema. Previously, metronidazole was available as a 0.75% gel. Improved solubility was achieved in a new, stable, aqueous gel that permitted the formulation of metronidazole 1.0%. This new formulation is highly spreadable, easy to use, cosmetically friendly, mild to the skin, nondrying, and moisturizing. The safety of metronidazole 1% gel was determined by the evaluation of its cumulative irritation, contact sensitization, phototoxicity, and photoallergy potential in healthy male and female patients. In this formulation, metronidazole was not irritating under occlusive application. Additionally, metronidazole 1% gel had a low potential for causing sensitization reactions, and no evidence suggested phototoxic or photoallergic reactions. PMID- 16706246 TI - A novel aqueous metronidazole 1% gel with hydrosolubilizing agents (HSA-3). AB - Rosacea is a common chronic dermatosis that often is characterized by the presence of facial erythema, visible blood vessels, papules, and pustules. Because the face is the predominant site of involvement, rosacea may cause serious psychologic trauma and can significantly affect the quality of life of individuals with the condition. The first topical therapy approved for rosacea by the US Food and Drug Administration was metronidazole for the treatment of inflammatory lesions and erythema. A stable aqueous gel formulation of 1% metronidazole has been developed with the novel combination of hydrosolubilizing agents. Metronidazole 1% gel is a clear gel that exhibits advantageous qualities. Some features of the new product include being highly spreadable, easy to use, cosmetically friendly, ultramild, nondrying, and moisturizing. Data are presented from a 21-day cumulative irritation study, an assessment of skin barrier function, an in vitro skin penetration study, an absorption following maximal topical exposure study, and a skin hydration study. PMID- 16706247 TI - The effect of Cetaphil Gentle Skin Cleanser on the skin barrier of patients with rosacea. AB - A good skin care regimen is a critical part of rosacea treatment; however, care must be taken to choose nonirritating products because individuals with rosacea tend to have sensitive skin, and irritants can trigger a worsening of symptoms. This study examines the use of Cetaphil Gentle Skin Cleanser (a nonalkaline nonirritating cleanser) in patients with rosacea. To eliminate the confounding effects of various treatments, the cleanser was studied for a 2-week period in the absence of rosacea therapy following a 2-week washout period in patients with mild to moderate rosacea. During the washout period, patients were asked to cleanse twice daily with Dove Sensitive Skin Bar. During the 2-week study period, patients were monitored for skin barrier function through transepidermal water loss (TEWL) and corneometry; patients also were monitored for rosacea severity. Thirty patients were enrolled. No significant increase in TEWL was demonstrated at any point during the study, indicating that the gentle skin cleanser did not damage the skin barrier. Additionally, the cleanser was shown to maintain skin hydration. Furthermore, a post hoc statistical analysis suggests there was a significant reduction (P<.05) in investigator-assessed rosacea severity on the cheeks, forehead, and nose at the end of week 1 and on the cheeks, forehead, and chin at the end of week 2 compared with the end of the washout period (after 2 weeks of cleansing with Dove Sensitive Skin Bar). The mild nonirritating action of the gentle skin cleanser was supported by the lack of adverse events and the tolerability shown in the study. There were no increases in erythema, scaling, dryness, stinging, burning, or lack of smoothness in the skin during the 2-week study period, despite the fact that patients were not being treated for their rosacea during the study. These results indicate that Cetaphil Gentle Skin Cleanser may be a good choice for the cleansing part of a total rosacea skin care regimen. PMID- 16706248 TI - AIDS, HIV, antiretrovirals and Africa. PMID- 16706249 TI - The current status of antiretroviral therapy in Africa. PMID- 16706250 TI - Are the best antiretrovirals being used in Africa? PMID- 16706251 TI - Logistics and models of implementing large-scale HIV treatment, care and support in Africa. PMID- 16706252 TI - 13th Conference on Retroviruses and Opportunistic Infections (CROI), Denver, USA, 5-8 February 2005. PMID- 16706253 TI - Stamp vignette on medical science. William F. Giauque--Nobel Prize for low temperature research. PMID- 16706254 TI - Thrombotic microangiopathy associated with malignant hypertension. PMID- 16706255 TI - Cardiac disease and stroke: will history repeat itself? PMID- 16706256 TI - Population-based study of the relationship between patent foramen ovale and cerebrovascular ischemic events. AB - OBJECTIVE: To determine whether patent foramen ovale (PFO) is a risk factor for a cryptogenic cerebrovascular ischemic event (CIE). METHODS: This case-control study of 1072 residents of Olmsted County, Minnesota, who underwent contrast transesophageal echocardiography between 1993 and 1997 included 519 controls without CIE randomly selected from the population, 262 controls without CIE referred for transesophageal echocardiography because of cardiac disease, 158 cases with incident CIE of obvious cause (noncryptogenic), and 133 cases with incident CIE of uncertain cause (cryptogenic). RESULTS: Large PFOs were detected in 108 randomly selected controls (20.8%), 22 referred controls (8.4%), 17 noncryptogenic CIE cases (10.8%), and 22 cryptogenic CIE cases (16.5%). After adjustment for age, sex, hypertension, smoking, atrial fibrillation, ischemic heart disease, and number of contrast injections, the presence of a large PFO was not significantly associated with group status (P=.07). Using the odds of the presence of large PFO in the randomly selected controls as the reference, the odds ratio (95% confidence interval) of the presence of large PFO was 0.47 (0.26 0.87) for referred controls, 0.69 (0.37-1.29) for noncryptogenic CIE cases, and 1.10 (0.63-1.90) for cryptogenic CIE cases. CONCLUSIONS: Patent foramen ovale is not a risk factor for cryptogenic ischemic stroke or transient ischemic attack in the general population. The PFO's importance in the genesis of cryptogenic CIE may have been overestimated in previous studies because of selective referral of cases and underascertainment of PFO among comparison groups of patients referred for echocardiography for clinical indications other than cryptogenic CIE. PMID- 16706257 TI - Population-based study of the relationship between atherosclerotic aortic debris and cerebrovascular ischemic events. AB - OBJECTIVE: To assess the validity of the suggestion that protruding atheromatous material in the thoracic aorta is an important cause of cerebrovascular ischemic events (CIEs) (ie, transient ischemic attack or ischemic stroke). METHODS: This case-control study of Olmsted County, Minnesota, residents who underwent transesophageal echocardiography (TEE) from 1993 to 1997 included controls without CIE randomly selected from the population, controls without CIE referred for TEE because of cardiac disease, cases with incident CIE of obvious cause (noncryptogenic), and cases with incident CIE of uncertain cause (cryptogenic). RESULTS: Of the 1135 subjects, 520 were randomly selected controls without CIE, 329 were controls without CIE referred for TEE, 159 were noncryptogenic CIE cases, and 127 were cryptogenic CIE cases. Complex atherosclerotic aortic debris in ascending and transverse segments of the arch was detected in 8 randomly selected controls (1.5%), 13 referred controls (4.0%), and 15 noncryptogenic (9.4%) and 4 cryptogenic (3.1%) CIE cases. After adjusting for age, sex, hypertension, smoking, atrial fibrillation, valvular heart disease, congestive heart failure, and atherosclerosis other than in the thoracic aorta, complex atherosclerotic aortic debris was not significantly associated with group status. With randomly selected controls as the referent group, odds ratios (95% confidence intervals) were 1.72 (0.61-4.87) for referred controls, 3.16 (1.18 8.51) for noncryptogenic CIE cases, and 1.39 (0.39-4.88) for cryptogenic CIE cases. CONCLUSIONS: Complex atherosclerotic aortic debris is not a risk factor for cryptogenic ischemic stroke or transient ischemic attack but is a marker for generalized atherosclerosis and well-established atherosclerotic and cardioembolic mechanisms of cerebral ischemia. Embolization from the aorta is not a common mechanism of ischemic stroke or transient ischemic attack. PMID- 16706258 TI - Paradoxical emboli in children and young adults: role of atrial septal defect and patent foramen ovale device closure. AB - OBJECTIVE: To describe a multicenter experience with patent foramen ovale (PFO) and atrial septal defect (ASD) device closure for presumed paradoxical emboli in children and young adults (<35 years old). PATIENTS AND METHODS: Medical records were reviewed of patients who had device closure of an ASD or PFO, who were younger than 35 years, and who had a history of presumed paradoxical embolus between January 1999 and August 2005 at Mayo Clinic, Rochester, Minn, University of Parma, Parma, Italy, and Loyola University Medical Center, Maywood, Ill. RESULTS: Forty-five patients fulfilled the inclusion criteria. Median patient age was 29.0 years (range, 5.0-34.9 years), and 23 patients (51%) were male. Clinical diagnoses included the following: stroke, 30 (67%); transient ischemic attack, 13 (29%); myocardial infarction, 1 (2%); and renal infarct, 1 (2%). Overall, 42 patients (93%) had a PFO, and 3 (7%) had an ASD. Seventeen patients had known cardiovascular disease risk factors: tobacco use (10 patients), hypercoagulable states (7 patients), systemic hypertension (3 patients), and hyperilpidemia (2 patients). No major procedural complications occurred. Median follow-up evaluation was performed at 5.3 months (range, 2.5-40.0 months). Forty-four patients (98%) had no recurrent neurologic events and no residual atrial shunt by contrast transthoracic echocardiography. CONCLUSIONS: Cryptogenic ischemic events occur in young patients and have serious sequelae. The potential for paradoxical embolization through a PFO or an ASD should be assessed in all such patients. In our short-term follow-up, device closure was a safe alternative therapeutic option for children and young adults with presumed paradoxical emboli. PMID- 16706259 TI - The role of pulmonary resection in small cell lung cancer. AB - OBJECTIVE: To analyze the outcome of surgical resection for patients with small cell lung cancer (SCLC). PATIENTS AND METHODS: We identified all patients who underwent thoracotomy for SCLC at our institution from January 1985 to July 2002. All patients were staged using the American Joint Committee on Cancer TNM system. RESULTS: The median age of the 77 patients (44 men and 33 women) was 65 years (range, 35-85 years). Operations performed included thoracotomy with biopsy of hilar mass in 10 patients, wedge excision in 30 (6 with talc pleurodesis), segmentectomy in 4, lobectomy in 28, bilobectomy in 3, and pneumonectomy in 2. Mediastinal lymphadenectomy was performed in 50 patients and lymph node sampling in 19. Postoperative therapy Included chemotherapy alone in 20 patients, radiation therapy in 3, and combined chemotherapy and radiation therapy in 40. Median tumor diameter was 4 cm (range, 1.0-10.0 cm). Postsurgical tumor stage was IA in 7 patients, IB in 11, IIA in 8, IIB in 7, IIIA in 30, IIIB in 10, and IV in 4. A total of 19 patients (25%) had complications: atrial arrhythmia in 7 patients, pneumonia in 6, prolonged air leak in 3, and myocardial infarction, postoperative bleeding, and cerebrovascular accident in 1 each. Operative mortality was 3% (2/77). Follow-up ranged from 4 days to 170 months (median, 19 months). At last follow-up, 20 patients were alive. The estimated overall 5-year survival was 27% when excluding the 10 patients who underwent a biopsy without additional surgery. Five-year survival for stage I and II combined (n=33) was 38% compared with only 16% for stage III and IV combined (n=34) (P=.02). Overall median survival was 24 months; median survival for patients who underwent curative surgery was 25 months compared with 16 months for those who had a palliative procedure (P=.34). CONCLUSION: Pulmonary resection in patients with stage I or stage II SCLC is safe with low mortality and morbidity. Curative resection is associated with long-term survival in early stage SCLC in some patients and should be considered in selected patients. PMID- 16706260 TI - The effect of adjuvant perfusion techniques on the incidence of paraplegia after repair of traumatic thoracic aortic transections. AB - OBJECTIVE: To analyze the effect of adjuvant perfusion techniques of the distal aorta on the outcome of traumatic thoracic aortic transections. PATIENTS AND METHODS: From 1973 to 2004, 72 patients (mean age, 39 years) with thoracic aortic transections arrived alive at the emergency department. Nineteen patients arrived in extremis and underwent emergency operations, 42 patients were stable and underwent diagnostic evaluation before surgery (4 patients experienced aortic rupture during evaluation), and 11 patients presented more than 24 hours after the accident. Sixteen patients died before aortic repair could be performed. Operative repair was possible in 53 patients (46 stable and 7 in extremis). Interposition graft was performed in 47 patients, and primary repair was performed in 6 patients. Morbidity, mortality, and paraplegia rate were analyzed. RESULTS: Patients in extremis had a mortality rate of 84% (16 of 19), stable patients had a mortality rate of 11% (4 of 38), patients who experienced rupture during evaluation had a mortality rate of 100% (4 of 4), and patients who underwent delayed operation had a mortality rate of 0% (0 of 11). The paraplegia rate with and without adjuvant distal aortic perfusion techniques was 2% (1 of 41 patients) and 33% (4 of 12 patients), respectively (P=.007). Mortality and paraplegia rates were 4% and 4% for partial bypass (n=24), 42% and 33% for the clamp and sew technique (n=12), 0% and 0% for Gott shunt (n=10), and 29% and 0% for full cardiopulmonary bypass (n=7), respectively. CONCLUSIONS: Although thoracic aortic transections remain a highly lethal injury, hemodynamically stable patients have a low operative mortality. Spinal cord injury is decreased by the use of adjuvant perfusion techniques that maintain distal aortic perfusion during cross-clamping of the aorta. PMID- 16706261 TI - A pen-and-paper coronary risk estimator for office use with patients with type 2 diabetes. AB - OBJECTIVE: To develop a pen-and-paper coronary heart disease (CHD) 10-year risk estimator for patients with type 2 diabetes based on the United Kingdom Prospective Diabetes Study (UKPDS) risk equation (based on 4000 patients with diabetes but only available electronically). PATIENTS AND METHODS: We used data collected from adults with type 2 diabetes from 6 primary care practices that participated in a randomized trial in Rochester, Minn; patients were enrolled in the study from July 2001 to December 2003, with follow-up through June 2004. We used multivariable linear regression of the CHD risk estimate to formulate prediction equations to estimate average (<15%), elevated, or high (>30%) 10-year CHD risk according to sex, age, diabetes duration, smoking, hemoglobin A1c level, systolic blood pressure, ratio of total cholesterol to high-density lipoprotein cholesterol, and microalbuminuria categories. We selected cut points for the predicted score, seeking to (1) maximize the number of patients with total agreement between our estimator and the UKPDS risk equation, (2) avoid any patient's risk being either overestimated or underestimated by 2 risk categories, and (3) overestimate rather than underestimate coronary risk. RESULTS: A total of 535 patients with type 2 diabetes participated in this study, 400 in the generation cohort and 135 in the validation cohort. Of the 400 patients in the generation cohort, our estimator had an 82% total agreement with the UKPDS calculation, 11% overestimated risk, and 7% underestimated UKPDS coronary risk (weighted kappa=0.77). Results were similar in the 135 patients in the validation cohort (kappa=0.79) and in an independent validation cohort of 52 patients attending a referral diabetes clinic (kappa=0.68). CONCLUSION: The pen-and-paper estimator facilitates the point-of-care estimation of coronary risk in situations in which use of a desktop or handheld version of the electronic UKPDS risk engine is not practical or feasible. In our experience, estimation of risk using this tool, when done with patients, can further patients' insight into their risk of coronary events, often leading to enlightened discussions about modification of Individual risk factors. PMID- 16706262 TI - Long-term care after percutaneous coronary intervention: focus on the role of antiplatelet therapy. AB - Arterial wall injury caused by percutaneous coronary intervention (PCI) triggers transient platelet activation and mural thrombosis; these effects are superimposed on the preexisting platelet hyperreactivity associated with underlying atherothrombosis. Platelet activation has been implicated in the major complications of PCI: acute and subacute thrombosis and restenosis. Antithrombotic and anticoagulant therapy minimizes thrombotic complications after PCI. Aspirin plus a thienopyridine (ticlopidine or clopidogrel) is more effective than aspirin plus heparin and extended warfarin therapy in preventing periprocedural ischemic events and subsequent stent thrombosis and results in less major and minor bleeding. Dual antiplatelet therapy with aspirin and clopidogrel (the preferred thienopyridine because of its superior hematologic safety) is recommended for at least 4 weeks to prevent subacute stent thrombosis with bare-metal stents and 3 to 6 months to prevent late-stent thrombosis with drug-eluting stents. Coronary atherothrombosis is a diffuse vascular disease, and reduction of the risk of future ischemic events requires strategies that extend beyond the focal treatment of stenotic lesions. Optimal long-term care after PCI requires aggressive systemic pharmacotherapy (antiplatelet agents, statins, beta blockers, and angiotensin-converting enzyme Inhibitors) in conjunction with therapeutic lifestyle changes (smoking cessation, weight reduction, dietary measures, and exercise). In this context, dual antiplatelet therapy (aspirin plus clopidogrel) is recommended for at least 12 months after PCI for prophylaxis of future atherothrombotic events. PMID- 16706263 TI - The physiology and potential clinical applications of ghrelin, a novel peptide hormone. AB - Ghrelin, a peptide hormone originally identified as the endogenous ligand of the growth hormone secretagogue receptor, is secreted primarily from the stomach and secondarily from the small intestine and colon. Ghrelin may also be expressed in the pancreatic islets, hypothalamus, pituitary, and several tissues in the periphery. The growth hormone secretagogue receptor is widely expressed, suggesting diverse physiologic roles for ghrelin. A growing body of evidence suggests that, in addition to its predictable effect on growth hormone secretion, ghrelin has an important role in the short-term regulation of appetite and the long-term regulation of energy balance and glucose homeostasis. Recent studies have implicated ghrelin in the regulation of gastrointestinal, cardiovascular, and immune function and have suggested a role for ghrelin in bone physiology. The identification of obestatin, a novel peptide hormone derived from the same gene as ghrelin, has recently added further complexity to ghrelin physiology. Obestatin appears to have actions opposite of ghrelin on energy homeostasis and gastrointestinal function. Despite the rapid progress, many questions remain unanswered, including the regulation of ghrelin and obestatin secretion, the downstream pathways that mediate their effects, and their precise physiologic endocrine and paracrine roles. This review presents data on ghrelin structure, expression, and function, with emphasis placed on human studies, highlighting areas that require future investigation and providing speculation about potential clinical applications of ghrelin agonists or antagonists. PMID- 16706264 TI - Diagnosis, screening, prevention, and treatment of osteoporosis. AB - Osteoporosis is the most common bone disease in humans and affects both men and women. The clinical and public health implications of the disease are substantial because of the mortality, morbidity, and cost of medical care associated with osteoporotic fractures. Osteoporosis is diagnosed on the basis of a low-impact or fragility fracture or low bone mineral density, which was best assessed by central dual-energy x-ray absorptiometry. Both nonpharmacological therapy (calcium and vitamin D supplementation, weight-bearing exercise, and fall prevention) and pharmacological treatments (antiresorptive and anabolic agents) may be helpful in the prevention and treatment of osteoporosis. Therefore, clinicians need to be vigilant in instituting primary prevention measures for those at high risk for osteoporosis and in instituting treatment for patients diagnosed as having the disease either by screening or a history of fracture. This article provides an overview of the diagnosis, screening, prevention, and treatment of osteoporosis. PMID- 16706265 TI - 21-year-old woman with flank pain. PMID- 16706266 TI - A practical approach to genetic testing for von Willebrand disease. AB - von Willebrand disease (vWD) is the most commonly diagnosed congenital bleeding disorder. The laboratory diagnosis of type 2 variants and type 3 vWD is reasonably well defined, and characterization of the von Willebrand factor (vWF) gene has facilitated definition of their molecular basis. However, for type 1 vWD, the laboratory diagnosis poses a diagnostic dilemma, and knowledge of its molecular basis is evolving. Characterization of the vWF gene and refinement of genetic techniques have led to an evolving repertoire of genetic tests. Genetic testing is costly, and thus judicious use will be increasingly important for appropriate genetic-counseling of patients with vWD and their family members. This article provides a practical approach to utilization of genetic testing in vWD. PMID- 16706267 TI - Intercostal lung cyst hernia in idiopathic pulmonary hemosiderosis (cyst necessitans). PMID- 16706268 TI - Monoclonal gammopathy of undetermined significance, Waldenstrom macroglobulinemia, AL amyloidosis, and related plasma cell disorders: diagnosis and treatment. AB - The spectrum of plasma cell disorders is broad. Monoclonal gammopathy of undetermined significance and smoldering multiple myeloma are asymptomatic disorders characterized by monoclonal plasma cell proliferation in the bone marrow in the absence of end-organ damage. Waldenstrom macroglobulinemia typically involves an ontogenically less mature lymphoplasmacytic bone marrow cell and is characterized by secretion of a monoclonal IgM protein. Solitary plasmacytoma is the only known potentially curable plasma cell disorder. Finally, AL (immunoglobulin light chain) amyloidosis and POEMS (polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin changes) syndrome are disorders characterized by low tumor burden but profound multisystemic disease. Updated diagnostic criteria for these disorders, risk stratification models to determine prognosis, and the current management of these diverse entitles are discussed in this review. PMID- 16706269 TI - How masked is the "masked peer review" of abstracts submitted to international medical conferences? PMID- 16706270 TI - From research evidence to context: the challenge of individualising care. PMID- 16706271 TI - Long-term retrospective study of 52 horses with sinunasal cysts. AB - REASONS FOR PERFORMING STUDY: The spectrum of clinical presentation, methods of diagnosis, management and prognosis in cases of sinunasal cysts (SNCs) requires more extensive study. Despite the extensive nature of SNCs, the prognosis for afflicted horses appears to have improved since earlier studies were conducted. OBJECTIVES: To evaluate prognosis and cosmetic outcome of surgical ablation in 52 cases of SNCs. METHODS: Fifty-two case records (1982-2005) of horses affected with SNCs were retrieved from the archives. Subject details, clinical signs, diagnostic techniques, surgical management and post surgical complications were extracted. Owners and referring veterinarians were contacted to assess the results of treatment. RESULTS: There was no biphasic age distribution. The major presenting signs in descending order of frequency were facial swelling, mucopurulent nasal discharge, nasal airflow obstruction and abnormal respiratory noise. Common endoscopic findings included narrowed nasal meati, a cyst in the nasal cavity and a cyst visible caudal to the nasal septum viewed from the contralateral unaffected nasal meatus. Typical radiological signs included a discrete mass in the sinunasal region, a diffuse increase in opacity over the sinunasal region, free fluid lines, nasal septal deviation, expansion of the ventral conchal sinus and distortion of dental apices. Cyst extirpation using an osteoplastic flap provided a successful outcome. Forty-five of 48 horses showed complete resolution of clinical signs after subtotal or total extirpation of the cyst wall. Follow-up information was not available for the other 3 horses. Thirty nine of 45 horses had a good to fair cosmetic appearance, judged to be good in 26 horses, fair in 13 and poor in 6. Results confirmed that the prognosis for full recovery has improved since a previous report of 15 cases. CONCLUSIONS AND POTENTIAL RELEVANCE: Equine sinunasal cysts may arise in horses of any age; presenting signs should alert clinicians to their likely presence. A discrete well-circumscribed mass found during radiographic examination provides supporting evidence of a cyst. The vivid yellow, translucent, seromucoid fluid aspirated from cysts is characteristic of the condition. PMID- 16706272 TI - Equine laminitis induced with oligofructose. AB - REASONS FOR PERFORMING STUDY: Experimental induction of equine laminitis with a reliable and clinically relevant model should facilitate understanding of the disease. Successful induction with oligofructose (OF) could link pasture consumption to laminitis. OBJECTIVES: To determine whether alimentary administration of OF induces laminitis. METHODS: Twelve horses were dosed with OF and 6 received a sham (placebo) treatment. Clinical observations were made and blood collected at 4 h intervals over a 48 h study period. Stained sections of the hoof wall lamellae, examined by light microscopy, were graded for laminitis severity. RESULTS: All horses administered OF, but no sham-treated controls, developed clinical and histological laminitis. CONCLUSIONS AND POTENTIAL RELEVANCE: Alimentary overload with OF is a valid induction model for studying the pathogenesis of laminitis. A link is therefore established between field cases of laminitis and pasture fructan content. PMID- 16706273 TI - Prevalence of gastric ulceration in Swedish Standardbreds in race training. AB - REASONS FOR PERFORMING STUDY: There is little information about the prevalence of gastric ulcers in Standardbred trotters and potential correlations between ulcers and various traits, e.g. age, sex, performance, temperament and feeding regimens, need further elucidation. OBJECTIVES: The prevalence of ulcers in the gastric squamous mucosa of Swedish Standardbreds was estimated and the associations between ulcer score and age, sex, performance, temperament and feeding regimens were determined. METHODS: Eighty Swedish Standardbreds in active race training, trained by 9 different trainers, underwent gastroscopy. Information on age, gender, status of training, performance over the last 4 months compared with the trainer's expectations before that period, behaviour, eating habits, feed type, bedding type, time spent outdoors per day and medical treatment during the last month was collected. Gastroscopy was performed and lesions in the squamous gastric mucosa were scored from 0 to 4 (0 = no lesions; 4 = > 5 lesions with haemorrhage, > 10 lesions with no haemorrhage, or a large area of diffuse loss of surface epithelium). RESULTS: Twenty-four (30%) of the horses had no lesions in the squamous mucosa, 22 (27.5%) had a score of 1; 21 (26.2%) score 2; 9 (11.2%) score 3; and 4 (5%) score 4. Horses that were in preparatory training and those that had raced during the last month were significantly more affected than horses that were fit for racing but had not raced during the last month, using a logistic regression model with trainer incorporated as a random effect. CONCLUSIONS: The study confirmed a high prevalence of ulcers in the gastric squamous mucosa of Standardbreds in race training. Of the studied parameters only status of training showed a significant association with gastric ulcers of the squamous mucosa. POTENTIAL RELEVANCE: Gastric ulceration is a common disease and diagnosis should be made by endoscopic examination of the gastric mucosa, since parameters such as age, gender, performance, behaviour, eating habits provide only weak clues as to which horses might be affected. Further studies are needed to determine the aetiology of the syndrome and to find ways to reduce, if possible, the frequency of gastric ulcers. PMID- 16706274 TI - Characterisation of lymphocyte subsets in the equine oviduct. AB - REASONS FOR PERFORMING STUDY: The equine oviduct is the site of fertilisation and location of embryonic development during the first 5 or 6 days. It therefore has an important influence on mare fertility. Although histopathological changes have been described previously, there is limited information regarding lymphocyte subtypes present in the mucosa of the normal equine oviduct. OBJECTIVES: To characterise the distribution of CD3+, CD4+, CD8+ and B lymphocytes in the equine oviduct from inseminated mares during oestrus and dioestrus, and from noninseminated mares during the immediate post ovulatory period. METHODS: Oviductal tissues were collected from noninseminated mares at oestrus (> 30 mm follicle, n = 4), at Day 1 post ovulation (n = 3) and at dioestrus (Day 7 post ovulation; n = 4). Oviducts were also collected from inseminated mares at Days 1, 2, and 3 post ovulation (n = 4 for each period). Cross-sections of tissues from the ampullar-isthmic junction from each oviduct were snap frozen and cryostat sections stained by the immunoperoxidase technique with monoclonal antibodies directed against equine lymphocyte surface markers for B cells as well as CD3+, CD4+ and CD8+ cells. RESULTS: In all oviductal sections examined, B cells were rare whereas T cells were relatively abundant. The predominant cell type found was the CD8+ phenotype, with a lesser number of CD4+ cells. Among mares, individual variation was large; therefore, although breeding status and stage of oestrous cycle appeared to alter lymphocyte populations, these differences were not significant. CONCLUSIONS AND POTENTIAL RELEVANCE: A population of CD3+, CD4+ and CD8+ cells exists within the mucosal region of the equine oviduct. The density of these cells is similar to that described in the human oviduct. Their function is not currently known, but they may be involved with modulation of the maternal response to the presence of spermatozoa or the early conceptus within the equine oviduct. As our capacity to differentiate these cell types improves, along with the ability to identify the specific cytokines they produce, their functional significance will become more apparent. PMID- 16706275 TI - Response to nasopharyngeal oxygen administration in horses with lung disease. AB - REASONS FOR PERFORMING STUDY: Guidelines for administration of oxygen to standing horses are unavailable because previous investigations of the efficacy of oxygen administration to increase arterial oxygenation in standing horses have produced equivocal results. OBJECTIVE: To determine the effect of nasal oxygen supplementation on inspired and arterial blood gas tensions in control horses and those with moderate to severe recurrent airway obstruction (RAO). METHODS: Normal horses (n = 6) and horses during an attack of RAO induced by stabling (n = 6) were studied. Oxygen was administered through either one or 2 cannulae, passed via the nares into the nasopharynx to the level of the medial canthus of each eye. Intratracheal inspired oxygen and carbon dioxide concentration and arterial blood gas tensions were measured at baseline and during delivery of 5, 10, 15, 20 and 30 l/min oxygen. RESULTS: Nasal cannulae and all but the highest oxygen flow rates were well tolerated. Fractional inspired oxygen concentration (F(I)O2) increased with flow but was significantly lower at all flow rates in horses with RAO compared with controls. Arterial oxygen tension (PaO2) was significantly increased (P < 0.001) by all flow rates, but was always lower in RAO-affected animals. At 30 l/min, PaO2 increased to 319 +/- 31 mmHg in control horses and 264 +/- 69 mmHg in horses with RAO. Additionally, a large arterial to end-tidal gradient for CO2 in RAO-affected horses was observed, indicating increased alveolar deadspace ventilation in these animals. CONCLUSIONS: The use of nasal cannulae to deliver oxygen effectively increases both F(I)O2 and PaO2 in horses with moderate to severe RAO. Oxygen flow rates up to 20 l/min are well tolerated, but flow rates of 30 l/min produce occasional coughing or gagging. POTENTIAL RELEVANCE: Oxygen therapy delivered by means of an intranasal cannula is a highly effective means of increasing arterial oxygen tension in horses with respiratory disease. Generally, flows of 10-20 l/min should be effective. If higher flows (20 30 l/min) are necessary, they should be delivered by means of 2 cannulae. PMID- 16706276 TI - Effective removal of equine arteritis virus from stallion semen. AB - REASONS FOR PERFORMING STUDY: A method of removing equine arteritis virus (EAV) from equine semen used for artificial insemination is urgently needed. Recent medical studies suggest that a double semen processing technique of density gradient centrifugation followed by a 'swim-up' can provide virus-free sperm preparations for assisted reproduction. OBJECTIVES: To investigate the use of the double semen processing technique to obtain virus-free sperm preparations from stallion semen containing EAV. METHODS: Aliquots of an ejaculate from an uninfected stallion were spiked with virus and processed by the double processing technique. The sperm preparations were tested by PCR for the presence of EAV. The procedure was repeated using an ejaculate from a known shedding stallion, testing processed and unprocessed aliquots by PCR and virus isolation. RESULTS: Virus free sperm preparations were obtained using the double sperm processing technique. The 'swim-up' step is apparently required to ensure complete virus removal. CONCLUSIONS: The double semen processing technique is potentially a useful and simple tool for the removal of EAV from the semen of shedding stallions. POTENTIAL RELEVANCE: The inclusion of density gradient centrifugation and 'swim-up' in protocols for the processing of semen for artificial insemination could help prevent the transmission of viral diseases carried in semen, such as EAV. PMID- 16706277 TI - The role of quantitative electromyography (EMG) in horses suspected of acute and chronic grass sickness. AB - REASONS FOR PERFORMING THE STUDY: Clinical evidence of motor neuron involvement in equine grass sickness (EGS) has not been reported. HYPOTHESIS: Quantitative electromyography (EMG) analysis can elucidate subtle changes of the lower motor neuron system present in horses with EGS, performed ante mortem. METHODS: Fourteen horses diagnosed clinically with acute, subacute or chronic EGS were examined and quantitative EMG performed. Previously published data on healthy horses and horses with proven lower motor neuron disease (LMND) were used as controls. In 8 horses post mortem examination was performed, and in 7 muscle biopsies of the lateral vastus muscle underwent histopathology and morphometry. RESULTS: Clinical electrophysiological evidence of neuropathy was present in 12 horses. Analysis of data from the first 4 horses resulted in 95% confidence intervals (CI) of nontransformed data for motor unit action potential (MUP) duration in subclavian, triceps and lateral vastus muscle of 11.0-13.7, 14.8-20.3 and 12.2-17.2 msecs, respectively, and for MUP amplitude 291-453, 1026-1892 and 957-1736 microV, respectively. For number of phases the 95% CI was 3.6-4.4, 2.9 3.6 and 2.9-3.4, respectively, and for number of turns 5.0-6.5, 4.3-5.3 and 3.7 4.6, respectively. No changes in duration of insertional activity were measured. Pathological spontaneous activity was observed in all horses. EGS as evidenced by degenerative changes in the autonomic ganglia in combination with minor degenerative changes of the spinal lower motor neurons was observed on post mortem examination in all 8 available autopsies. In muscle biopsies of 4 out of 7 horses changes consistent with slight neurogenic atrophy were found. CONCLUSIONS AND POTENTIAL RELEVANCE: EMG results demonstrated the presence of a neuropathy of skeletal muscles in all horses suspected to have EGS. The combination of clinical and electrophysiological evidence may aid differential diagnosis of neurogenic disease in cases of weight loss and colic. PMID- 16706278 TI - Antagonism of detomidine sedation in the horse using intravenous tolazoline or atipamezole. AB - REASONS FOR PERFORMING STUDY: The ability to shorten the duration of sedation would potentially improve safety and utility of detomidine. OBJECTIVES: To determine the effects of tolazoline and atipamezole after detomidine sedation. HYPOTHESIS: Administration of tolazoline or atipamezole would not affect detomidine sedation. METHODS: In a randomised, placebo-controlled, double-blind, descriptive study, detomidine (0.02 mg/kg bwt i.v.) was administered to 6 mature horses on 4 separate occasions. Twenty-five mins later, each horse received one of 4 treatments: Group 1 saline (0.9% i.v.) as a placebo control; Group 2 atipamezole (0.05 mg/kg bwt i.v.); Group 3 atipamezole (0.1 mg/kg bwt i.v.); and Group 4 tolazoline (4.0 mg/kg bwt i.v.). Sedation, muscle relaxation and ataxia were scored by 3 independent observers at 9 time points. Horses were led through an obstacle course at 7 time points. Course completion time was recorded and the ability of the horse to traverse the course was scored by 3 independent observers. Horses were videotaped before, during and after each trip through the obstacle course. RESULTS: Atipamezole and tolazoline administration incompletely antagonised the effects of detomidine, but the time course to recovery was shortened. CONCLUSIONS AND POTENTIAL RELEVANCE: Single bolus administration of atipamezole or tolazoline produced partial reversal of detomidine sedation and may be useful for minimising detomidine sedation. PMID- 16706279 TI - Use of transabdominal ultrasonography to determine the location of cryptorchid testes in the horse. AB - REASONS FOR PERFORMING STUDY: Transrectal ultrasonography is a reliable technique to identify intra-abdominal testes, but can be hazardous or impractical in fractious, juvenile or small equids. The transabdominal approach therefore requires validation. OBJECTIVES: To evaluate the sensitivity and specificity of transabdominal ultrasonography to localise cryptorchid testes. METHODS: Thirty eight horses admitted for cryptorchidism underwent transabdominal ultrasonography to locate errant testes. Location of the testes was confirmed during surgery (n = 37) or necropsy (n = 1). RESULTS: Horses weighed 175-760 kg. Twenty-two testes were located within the abdomen, 19 in the inguinal space and 3 in a subcutaneous location. In 2 horses, a thick winter coat prevented adequate contact between the probe and the skin, and visualisation of their inguinal testis and one abdominal testis was missed during transabdominal ultrasonography. Excluding the 2 horses with excessive hair, the sensitivity of transabdominal ultrasonography to locate errant testes was 97.6% (93.2% when all horses were included), and its specificity 100%. CONCLUSIONS: Transabdominal ultrasonography is a reliable, safe and immediate technique to diagnose cryptorchidism precisely in horses of all ages, sizes and temperaments. POTENTIAL RELEVANCE: Transabdominal ultrasonography provides an immediate definitive diagnosis of adominal cryptorchidism and should enhance selection of an appropriate surgical approach for their removal. PMID- 16706280 TI - Minimally invasive plate fixation of lower limb injury in horses: 32 cases (1999 2003). AB - REASONS FOR PERFORMING STUDY: Complications of osteosynthesis including superficial and deep infections continue to affect adversely outcome in equine surgery of the lower limb. OBJECTIVE: To describe a technique for minimally invasive plate fixation in horses and evaluate clinical results of the technique. HYPOTHESIS: Lower complication rates would occur in horses treated with a minimally invasive approach compared with conventional techniques. METHODS: Cases of minimally invasive plate fixation from January 1999 to December 2003 were evaluated. Case records, radiographs and race records for horses that sustained distal third metacarpal/metatarsal (Mc/tIII) condylar fractures were assessed. Follow-up information was obtained and results analysed. In addition, results for horses that received arthrodesis by a conventional, open approach during the same period were evaluated and statistical analysis was performed. RESULTS: Thirty-two cases of minimally invasive plate fixation were identified (22 incomplete distal Mc/tIII condylar fractures with a spiralling or diaphyseal component, 10 metacarpophalangeal arthrodeses and 4 pastern arthrodeses). For both types of arthrodesis, surgery times and infection rates were lower and survival rates higher in the minimally invasive group, but no statistical difference was found. Spiralling condylar fractures were successfully managed, without catastrophic fracture. CONCLUSIONS AND POTENTIAL RELEVANCE: Minimally invasive percutaneous plate osteosynthesis (MIPPO) can be successfully used for select applications in horses, and may reduce morbidity and mortality in equine fracture repair. PMID- 16706281 TI - Quantification by real-time PCR of the magnitude and duration of leucocyte associated viraemia in horses infected with neuropathogenic vs. non neuropathogenic strains of EHV-1. AB - REASONS FOR PERFORMING STUDY: Neurological disease in horses caused by infection with certain 'paralytic' strains of equine herpesvirus-1 (EHV-1) is a potentially devastating condition the pathogenesis of which is poorly understood. Preliminary observations in both experimentally induced and naturally occurring cases of the central nervous system disease have revealed a more robust cell-associated viraemia in horses infected with paralytic isolates of EHV-1, relative to horses infected with abortigenic isolates. To investigate further this pathogenesis relevant question, the present study was performed using a greater number of horses and a more precise method for quantification of EHV-1 DNA present in viraemic leucocytes. OBJECTIVE: To compare the magnitude and duration of leucocyte-associated viraemia in seronegative, age-matched foals following infection with paralytic vs. abortigenic isolates of EHV-1. METHODS: Peripheral blood mononuclear cells (PBMC) were collected from 20 weanling foals at 2, 4, 7, 9, 11, 14 and 21 days after intranasal inoculation with either paralytic or abortigenic isolates of EHV-1. The amount of EHV-1 DNA present in each PBMC sample was measured by real-time quantitative PCR. RESULTS: Foals inoculated with paralytic strains of EHV-1 developed both a greater magnitude and longer duration of PBMC-associated viraemia than foals inoculated with abortigenic strains of the virus. CONCLUSIONS: Both the higher magnitude and longer duration of cell associated viraemia contribute to the risk for development of neurological signs in horses infected with paralytic strains of EHV-1. POTENTIAL RELEVANCE: Our results provide empirically derived, scientific data that contributes to a better understanding of the pathogenetic basis for the differing abilities of paralytic and abortigenic strains of EHV-1 to cause post infection central nervous system disease in the horse. The findings identify the importance of minimising the quantitative burden of viraemic leucocytes that follows exposure to the virus, by the use of effective therapeutic antiviral drugs and efficacious prophylactic vaccines that stimulate cytotoxic immune responses against EHV-1 infected cells. PMID- 16706282 TI - Shoeing sound warmblood horses with a rolled toe optimises hoof-unrollment and lowers peak loading during breakover. AB - REASONS FOR PERFORMING STUDY: Overload injuries in sport horses commonly occur; shoeing techniques are believed to be important in prevention of these injuries, but there is a paucity of scientific information identifying the potential connection. OBJECTIVES: To test a horseshoe with a modified rolled toe designed to ease the process of breakover and decrease loading of lesion-prone structures of the distal limb. METHODS: Twenty clinically sound Warmblood horses trotted over a track containing a pressure/force measuring system and 6 infrared cameras. The horses were measured with 2 types of shoes, standard flat shoes and shoes with a rolled toe. The shoeing procedure was randomised and horses had 2 days between measurements to adapt to the shoes. RESULTS: Limb placement and timing characteristics, e.g. breakover duration, did not change significantly. There was an improvement in the ease of movement to roll over the toe in the shoes with a rolled toe, due mainly to a smoother hoof-unrollment pattern. The peak indicative moment decreased substantially at the onset of breakover in the shoe with the rolled toe. CONCLUSIONS: With a rolled toe the process of hoof-unrollment is smoother, which improves the coordination of this process, and lowers peak loading of the distal limb during breakover. POTENTIAL RELEVANCE: This study stresses the importance of proper shoeing in sound horses, showing that shoe modifications can optimise the loading characteristics of the distal limb and therefore might be a means to prevent sport horses from overload injuries. PMID- 16706283 TI - Comparison of concentrations of Rhodococcus equi and virulent R. equi in air of stables and paddocks on horse breeding farms in a temperate climate. AB - REASONS FOR PERFORMING STUDY: Rhodococcoccus equi is a significant cause of bronchopneumonia in foals worldwide. Infection of the lungs is believed to result from inhalation of virulent R. equi in dust from contaminated environments. A measure of infectious risk in an environment is the level of airborne contamination. OBJECTIVES: To assess and compare the level of airborne virulent R. equi in paddocks and stables. METHODS: Air samples were collected sequentially over the 2003 foaling season from the paddocks and stables on 3 Irish horse breeding farms affected by R. equi pneumonia. Colony blotting and DNA hybridisation techniques allowed quantitation of virulent R. equi. RESULTS: The odds of detecting airborne virulent R. equi in stables were 173 times greater than in paddocks. The median airborne concentration of virulent R. equi was significantly higher (P < 0.001) in stables than in paddocks on all farms. These observations suggested that stables were high-risk areas for infection. CONCLUSIONS AND POTENTIAL RELEVANCE: Our results indicate that contaminated stables are a significant risk factor in the epidemiology of R. equi pneumonia on horse-breeding farms in a temperate climate, such as in Ireland. Management strategies that improve the air hygiene of stables, through better ventilation, use of less fragile bedding material and the use of fogging agents to reduce the airborne concentration of virulent R. equi, may reduce the incidence and severity of R. equi pneumonia on farms. PMID- 16706284 TI - A technique for pelvic radiography in the standing horse. AB - REASONS FOR PERFORMING STUDY: An alternative technique of radiographing the pelvis in the standing horse is required, to avoid the risks associated with general anaesthesia. HYPOTHESIS: That lateral oblique radiography in the standing horse would be a useful technique in the investigation of pelvic injury. OBJECTIVES: To describe the technique of lateral oblique pelvic radiography in the standing horse and demonstrate the feasibility and usefulness of this technique. METHODS: A technique for lateral oblique radiography in the standing horse was devised and retrospective review made of radiographic findings in 18 clinical cases. RESULTS: The caudal iliac shaft, greater trochanter of the femur, femoral head, acetabulum and coxofemoral articulation on the side under investigation were visualised consistently using this technique. Of the 18 cases, 3 iliac shaft fractures, 1 acetabular fracture, 2 coxofemoral luxations and 4 horses with new bone formation around the coxofemoral joint and/or proximal femur were identified. CONCLUSIONS: Lateral oblique radiography in the standing, conscious horse can be used to investigate conditions affecting the caudal iliac shaft, coxofemoral articulation and proximal femur in the horse. POTENTIAL RELEVANCE: The technique is straightforward, noninvasive and useful in the investigation of horses with suspected pelvic injury. However, not all pelvic injuries would be identified, and normal radiographic findings do not rule out injury or fractures elsewhere in the pelvis. PMID- 16706285 TI - Long-term follow-up on a large number of horses that underwent transarterial coil embolisation (TCE) for guttural pouch mycosis (GPM). PMID- 16706286 TI - Myopathy in American miniature horses. PMID- 16706287 TI - Haemangiosarcoma involving the third eyelid in the horse: a case series. PMID- 16706288 TI - Pathogenesis of equine herpesvirus-associated neurological disease: a revised explanation. PMID- 16706289 TI - Fracture resistance of inlay-retained fixed partial dentures reinforced with fiber-reinforced composite. AB - In this study, the effect on the fracture load of inlay-retained composite fixed partial dentures (FPDs) caused by reinforcing them with fiber-reinforced composite (FRC) in different positions was examined. Experimental FPDs were fabricated using Estenia/EG Fiber (Kuraray Medical). Pontic reinforcement was then performed in one of the following three ways: reinforced the central area in a single line or in double straight lines, or reinforced the bottom in a curved line. The finding was that, when the area ranging from the connector to the bottom of the pontic was reinforced with FRC in a curved line, the fracture load of the FPDs tended to become higher. In addition, the FPDs fractured mainly at the veneering composite of the connector area. Based on the results of this study, it was concluded that reinforcement using FRC is effective, and that the veneering composite in the connector area needs to have sufficient strength to prevent the fractures. PMID- 16706290 TI - Shear bond strength of tooth-colored indirect restorations bonded to mid-coronal and cervical dentin. AB - To investigate the bonding of tooth-colored indirect restorations to cervical dentin, shear bond strengths of bonded resin inlays to cervical dentin and mid coronal dentin were investigated and compared. Indirectly fabricated composite inlays (Estenia) were cemented with a dual-cured resin cement (Panavia Fluoro Cement II) to either cervical or mid-coronal dentin. Resin cement was cured with or without light irradiation for 20 seconds. After 24-hour or one-week storage, the bonded inlays were subjected to a micro-shear bond test. The light-cure method showed significantly higher bond strengths to both dentin regions compared with the self-cure method. Further, significant improvement in bonding after one week storage was found in the case of light-cure method rather than the self-cure method. Although the cervical dentin tended to show lower bond strength than mid coronal dentin, significant difference between the two dentin regions was only found when the resin cement was light-cured and stored for only one day. PMID- 16706291 TI - Abrading increases oxygen and hardness of titanium surface. AB - CP Ti was mirror-polished and then abraded with waterproof SiC papers of two different grit sizes: 16 and 3 microm. As-polished and abraded surfaces were characterized by means of EPMA, XPS, XRD, and hardness test. Oxygen in the mirror polished surface was uniformly distributed at the lowest level. Comparatively, abrading with SiC papers increased the surface oxygen amount and hardness. Owing to its excellent abrasivity, the coarse grit efficiently scratched the surface and hindered the regenerated oxide film from growing thick, but allowed only the metal-oxide interfacial gradient zone to extend. But, the fine grit merely rubbed the surface and allowed both the oxide film and interfacial zone to extend. Further, the surface appeared to be lightly yellow-colored, suggesting that the oxide film was thicker, probably within 10 nm, than the nominal one. When compared with the bulk, the interfacial zone was rich in oxygen and therefore subjected to high coherency strain, which was introduced to relieve the great lattice mismatch between the outer and inner layers of titanium substrate. Effects of solute oxygen hardening and strain hardening were speculated to be responsible for the surface hardening of both SiC-abraded surfaces. In conclusion, abrading with a coarse grit led to accumulation of a high, non uniform strain in the titanium substrate, thereby hardening the surface further. PMID- 16706292 TI - Influence of abutment materials on the resultant color of heat-pressed lithium disilicate ceramics. AB - The purpose of this study was to evaluate the influence of abutment materials on the color of IPS Empress 2 ceramic coping with different thicknesses. Ceramic coping specimens (12.0x12.0x0.8-2.0 mm) were fabricated from IPS Empress 2 material (Ingot-100, n=5/group). Abutment specimens were fabricated from a build up composite, a gold alloy, or a silver-palladium alloy. Color was evaluated using a colorimeter according to the CIE L*a*b* system. The L*a*b* values of the ceramic coping specimens of different thicknesses on each abutment specimen were measured. Following which, the color difference (deltaE*ab) values between the ceramic coping specimens on various abutment specimens were calculated. Significant differences in deltaE*ab value were observed among different abutment specimens at certain ceramic coping thicknesses (P<0.05). Thus, it was concluded that the color of IPS Empress 2 coping material was influenced significantly by both the thickness of the coping and the color of the abutment material. PMID- 16706293 TI - Effect of enamel shades on color of layered resin composites. AB - The purpose of this study was to evaluate the effect of the covering enamel layer of various thicknesses on the color of layered resin composites (LRC). To this end, the CIELAB parameters of the following were evaluated: underlying base (ULB: 2.0 mm of body shade + 3.0 mm of opaque shade), inherent color of the enamel shade (4.0 mm of enamel shade + ULB), and LRC disks (0.5, 1.0, 2.0, 3.0 mm of enamel shade + ULB) of Filtek Supreme (FS, 3M) and Gradia Direct (GD, GC). To assess the effect of the enamel layer on the color of LRC, color difference between ULB and LRC was calculated. With FS, the enamel layer had no major effect on the color of LRC--regardless of thickness. With GD, the enamel layer had a significant effect on the color of LRC, even if the thickness was only 0.5 mm. The difference in enamel layer effect between the two products could be attributed to the difference in color characteristics of the two enamel shades. PMID- 16706294 TI - Hydroxyapatite film coating by thermally induced liquid-phase deposition method for titanium implants. AB - Thermally induced liquid-phase deposition method was employed to produce a thin hydroxyapatite (HA) film on a titanium substrate in a metastable calcium phosphate solution. Titanium foil of 100x10x0.02 mm3 was used as a substrate. Prior to HA coating, the substrate was immersed in 5 M NaOH solution at 60 degrees C for 24 hours. Substrate temperature was kept constant at 60 degrees C for 0.5-3 hours by electrical heating with a DC power source in the metastable calcium phosphate solution. An X-ray diffractogram indicated that the film deposited on the titanium substrate was composed of HA. The amount of HA deposited increased with increase in heating time. These results suggested that a uniform HA film can be formed by simple chemical and thermal treatments. Based on the results of this study, this technique seemed to be useful for producing uniform HA coatings on complex-shaped and porous dental implants. PMID- 16706295 TI - The effect of different solutions on the bond strength of soft lining materials to acrylic resin. AB - In this study, we investigated the effect of different solutions--coffee, tea, Turkish coffee, artificial saliva, mouthwash, denture cleanser, and distilled water--on the bond strength of soft lining materials (one acrylic- and three silicone-based) to acrylic resin. Acrylic specimens (40x10x10 mm) were prepared for the bond strength test. The specimens were stored in different solutions and tested after 24 hours, seven days, and 30 days. Using analysis of variance, the bond strength of soft lining materials to acrylic resin was found to be related to the type of material, storage time, and storage solution. Visco-gel had the lowest bond strength to cured acrylic resin, where its strength ranged from 0.149 MPa to 0.784 MPa. The bond strength of Visco-gel was also found to increase with time. Molloplast-B and Ufi Gel P demonstrated better bond strength than Visco-gel and Mollosil. PMID- 16706296 TI - The effect of bonding system and composite type on adaptation of different C factor restorations. AB - This study evaluated the effect of two adhesive systems and four resin-based composites on the marginal sealing and cavity wall adaptation of restorations with different C-factors. Cylindrical cavities, 1 mm deep and 3 mm in diameter (C factor=2.3) or 2 mm in diameter (C-factor=3), were prepared on superficial bovine dentin surfaces. The teeth were restored with Clearfil SE Bond or Single Bond adhesive system followed by hybrid (Photo Clearfil Bright or Z100) or flowable (Filtek Flow or Estelite LV) resin composite. After thermocycling, a dye penetration test was carried out to evaluate the degree of marginal leakage and cavity-wall gap formation. Cavity-wall gap formation increased when the C-factor increased from 2.3 to 3, except for Estelite LV resin composite. In terms of marginal sealing, Clearfil SE Bond showed better performance than Single Bond. When using Clearfil SE Bond, flowable composites improved resin composite adaptation to the cavity wall compared with hybrid composites. PMID- 16706297 TI - Effects of sintering temperature over 1,300 degrees C on the physical and compositional properties of porous hydroxyapatite foam. AB - Porous hydroxyapatite (HAP) foam permits three-dimensional (3D) structure with fully interconnecting pores as well as excellent tissue response and good osteoconductivity. It is therefore thought to be a good candidate as scaffold material for bone regeneration and as a synthetic bone substitute material. To fabricate better porous HAP foam, improved physical and structural properties as well as higher osteoconductivity are desired. In the present study, the effects of sintering temperature on the physical and compositional properties of porous HAP foam were evaluated by employing high sintering temperature starting at 1,300 degrees C up to 1,550 degrees C. The mechanical strength of porous HAP foam increased with sintering temperature to reach the maximum value at 1,525 degrees C, then decreased slightly when sintering temperature was further increased to 1,550 degrees C. Alpha tricalcium phosphate (alpha-TCP) was formed, and thus the porous HAP foam became biphasic calcium phosphate. Biphasic calcium phosphate consisting of both alpha-TCP and HAP had been reported to show higher osteoconductivity than HAP alone. We therefore recommend 1,500-1,550 degrees C as the sintering temperature for porous HAP foam since this condition provided the most desirable physical properties with biphasic calcium phosphate composition. PMID- 16706298 TI - Shear bond strengths of self-etching adhesives to caries-affected dentin on the gingival wall. AB - The purpose of this study was to evaluate the bonding ability of five current self-etching adhesives to caries-affected dentin on the gingival wall. Seventy extracted human molars with approximal dentin caries were employed in this study. In order to obtain caries-affected dentin on the gingival wall, grinding was performed under running water. Following which, specimens mounted in acrylic blocks and composite resins of the bonding systems were bonded to dentin with plastic rings and then debonded by shear bond strength. With Clearfil SE Bond, bonding to caries-affected dentin showed the highest bond strength. With Optibond Solo Plus Self-Etch, bonding to caries-affected dentin showed higher shear bond strength than AQ Bond, Tyrian SPE & One-Step Plus, and Prompt-L-Pop (p<0.05). Further, the bond strengths of Clearfil SE Bond and Optibond Solo Plus Self-Etch to sound dentin were higher than those of Prompt-L-Pop, AQ Bond, and Tyrian SPE & One-Step Plus (p<0.05). In conclusion, besides micromechanical interlocking through hybrid layer formation, bond strength of self-etch adhesives to dentin may be increased from additional chemical interaction between the functional monomer and residual hydroxyapatite. The results of this study confirmed that differences in bond strength among self-etching adhesives to both caries-affected and sound dentin were due to chemical composition rather than acidity. PMID- 16706299 TI - Effect of a caries-detecting solution on the tensile bond strength of four dentin adhesive systems. AB - This study investigated the effect of a caries-detecting solution on the tensile bond strength (TBS) to sound bovine dentin--which was either rinsed thoroughly of or contaminated with the caries-detecting solution. Caries Detector (1.0% acid red in propylene glycol) was applied on flat dentin surfaces for 10 seconds, rinsed, and dried with syringe air. In another group, Caries Detector was not rinsed but air-dried. Then, the surfaces were treated with one of the following adhesive systems: Clearfil Protect Bond, Clearfil SE Bond, One-Up Bond F, or Single Bond. Furthermore, an ingredient of Caries Detector, either 1.0% acid red aqueous solution or propylene glycol, was applied to evaluate the effect of each component. In the control groups, Caries Detector was not applied to the dentin surfaces. Finally, a resin composite was light-cured and the TBS measured. Fractured specimens and treated dentin surfaces were observed by SEM. Caries Detector did not reduce the tensile bond strength of any adhesive system (p>0.05) when rinsed thoroughly. On the other hand, when dentin surface was contaminated with Caries Detector, TBS decreased significantly with Clearfil SE Bond and Single Bond. As for the ingredients of Caries Detector, the effect of acid red on TBS was not significant, but that of propylene glycol was significant. PMID- 16706300 TI - Effect of adhesive primer developed exclusively for heat-curing resin on adhesive strength between plastic artificial tooth and acrylic denture base resin. AB - Despite progress in the development of denture base resin and artificial tooth materials, dental clinics are still plagued with artificial teeth falling off the denture base--due to poor bond strength--after denture delivery. Against this background, this study sought to examine the effect and durability of an adhesive primer developed exclusively for heat-curing resin on the adhesive strength of heat-curing denture base acrylic resin to plastic artificial tooth. Test specimens were divided into four groups according to the treatment method of the artificial tooth's test bonding surface: air abrasion, adhesive primer application, adhesive primer application after air abrasion, and pretreatment only (control). After heat curing of acrylic resin onto the bonding surface, shear test was performed for two storage periods: 24-hour versus 100-day water storage. From the results obtained, it was revealed that the evaluated adhesive primer was significantly effective in increasing adhesive strength between artificial tooth and acrylic resin, although specimens were stored in water for 100 days. PMID- 16706301 TI - Bactericidal effect of Er,Cr:YSGG laser on Streptococcus mutans. AB - The aim of this study was to compare the antibacterial activities of Er,Cr:YSGG laser with two different power outputs against a chlorhexidine gluconate-based cavity disinfectant. A cavity tooth model test was used to determine the antibacterial activity. Four cylindrical cavities were prepared on the dentin surface of 10 bovine incisors and left in contact with Streptococcus mutans for 72 hours to allow bacterial invasion. Following which, Er,Cr:YSGG laser with 0.75 W and 1 W power outputs and a chlorhexidine gluconate-based cavity disinfectant were applied separately on one of the three infected cavities, whereas the fourth was left untreated for control. Standardized amounts of dentin chips were obtained from the cavity walls, and the number of bacteria recovered was counted. Statistical analysis was carried out using one-way ANOVA and Dunnett's C test (p=0.05). No significant differences were observed among the data obtained from the chlorhexidine gluconate-based cavity disinfectant and the two Er,Cr:YSGG laser groups (p>0.05). However, when compared to the control group, both Er,Cr:YSGG laser groups and the chlorhexidine gluconate-based cavity disinfectant resulted in significantly less bacterial recovery (p<0.05). In conclusion, the antibacterial activity on S. mutans demonstrated by Er,Cr:YSGG laser with both energy outputs was similar to that of the tested chlorhexidine gluconate-based cavity disinfectant. PMID- 16706302 TI - Effects of different sizes of beta-tricalcium phosphate particles on bone augmentation within a titanium cap in rabbit calvarium. AB - This study evaluated the effects of different sizes of beta-TCP particles on bone augmentation within a titanium cap. In 20 rabbits, the calvarium was exposed and a circular groove was prepared. After marrow penetration, a standardized hemispherical titanium cap was placed in the circular grove. The cap was filled with small-sized (100-250 microm) or medium-sized (250-500 microm) beta-TCP particles for the experimental site and without beta-TCP for the control site. After one and three months of healing, the animals were euthanized and examined histologically. There was a statistically significant difference in the amount of mineralized bone generated between the experimental and control groups in the three-month specimens. Furthermore, the medium-sized particles showed significantly more mineralized bone than did the small-sized particles. Based on these findings, we suggested that beta-TCP might be effective for bone formation and that medium-sized particles are more useful than small-sized particles in bone maturation. PMID- 16706303 TI - Coronal leakage inhibition in endodontically treated teeth using resin-coating technique. AB - The purpose of this study was to evaluate coronal leakage inhibition using a resin-coating technique after endodontic treatment. Thirty-six human incisors were cut at the cementoenamel junction, and endodontic obturations were performed. The specimens were randomly divided into three groups according to post space preparation (10 mm depth), resin coating, and temporization. For the resin coating, the dentin surface was coated with either a combination of Clearfil SE Bond and Protect Liner F (SE/PLF) or RZII (RZ). Then, the specimens were stored in 37 degrees C distilled water for 24 hours and placed in 1% methylene blue solution for 48 hours. After which, the specimens were sectioned faciolingually along the root canal and the length of dye penetration was measured from the cementoenamel junction. Three-way ANOVA revealed that the dye penetration scores were influenced by post space preparation, resin coating, and temporization. Resin coating with RZ significantly reduced the dye penetration score and SE/PLF completely eliminated dye penetration. PMID- 16706304 TI - Corrosion behavior of pure titanium and titanium alloys in various concentrations of Acidulated Phosphate Fluoride (APF) solutions. AB - The corrosion behaviors of Ti, Ti-6Al-7Nb and Ti-6Al-4V alloys, and an experimentally produced Ti-0.5Pt alloy in 0.05% to 2.0% concentrations of Acidulated Phosphate Fluoride (APF) solutions (corresponding to 226 to 9,050 ppm fluoride at pH 3.5 or 3.6) were examined. While the corrosion of Ti, Ti-6Al-7Nb and Ti-6Al-4V alloys might occur easily even in a diluted 0.05% APF solution, dissolution of Ti from the Ti-0.5Pt alloy was observed only in test solutions with APF concentration exceeding 0.2%. When Ti-6Al-7Nb and Ti-6Al-4V alloys were immersed in 2.0% APF solution, their surfaces were entirely covered by compact corrosion products of Na3TiF6 due to severe corrosion. As a result, Ti dissolution was prevented and the amount of Ti dissolved decreased. However, since Ti was covered by porous, large-sized corrosion products of Na3TiF6 and that Ti-0.5Pt alloy was not covered with any corrosion product, the amount of Ti dissolved increased in the 2.0% APF solution. PMID- 16706305 TI - Mineral concentration of natural human teeth by a commercial micro-CT. AB - This study aimed to evaluate a commercial micro-CT system (microCT 20) for quantitative analysis of mineral concentration in human enamel and dentin using different methodologies, and thereby compare the obtained results with established data from published literature. A micro-CT device set at 50 kVp (160 microA) was used to scan five whole molars (G1) and five molars ground to 6-mm thickness (G2), as well as evaluate the mineral concentration of the samples. Mean mineral contents for enamel and dentin were 2.57 (+/- 0.12) and 1.53 (+/- 0.12) g/cm3 for G1, and 2.76 (+/- 0.03) and 1.45 (+/- 0.02) g/cm3 for G2. Difference between the groups was significant for enamel. For dentin, there was a clear although not significant tendency towards higher values with G1. The equipment could identify and differentiate a higher mineral content of the tooth enamel and dentin from the external to the inner tissue. Further, the absolute mean values of mineral concentration were lower in whole tooth samples than in sectioned samples due to beam hardening. In conclusion, the equipment is well suited for quantifying the mineral content of teeth. However, it is necessary to consider the limited acceleration voltage of the microCT 20 system and to limit sample evaluation to 6-mm thickness. PMID- 16706306 TI - Effect of 4-MET- and 10-MDP-based primers on resin bonding to titanium. AB - The purpose of this study was to investigate the effect of a 4-MET- and 10-MDP based primer on the bond strength of two resin cements (SuperBond C&B, Sun Medical; Panavia Fluoro Cement, Kuraray) to titanium (Ti). Ti plates were treated with six experimental primers consisting of, respectively, 10-MDP and 4-MET in concentrations of 0.1, 1 and 10wt%, or were kept untreated (control). The highest tensile bond strength of Panavia Fluoro Cement to Ti was obtained when the Ti surface was pre-treated with 10wt% 10-MDP and was significantly higher than that when a lower concentrated 10-MDP-based primer or any 4-MET-based primer was used. On the contrary, no significant difference in tensile bond strength of SuperBond C&B was found for the untreated and six pre-treated Ti surfaces, although pre treatment with each 10-MDP-based primer resulted in a higher tensile bond strength as compared to any 4-MET pre-treatment. Altogether, the data obtained strongly suggest that 10-MDP is effective to improve the adhesive performance of resin to titanium. PMID- 16706307 TI - Staining of hybrid composites with coffee, oolong tea, or red wine. AB - This study examined the surface staining mechanism of a photopolymerized composite by coffee, oolong tea, and red wine. Dental composite was subjected to an experimental 24-hour staining cycle: 17-hour immersion in artificial saliva solution containing 0.3% mucin followed by 7-hour immersion in coffee, tea, or wine. After one, two, and four weeks, digital images of the composite surface were analyzed in grayscale mode with an imaging analyzer. Specimens polished but not immersed were used as a baseline measurement for color change. Additionally, the effects of mechanical brushing and chlorhexidine on drink-induced staining were examined. Wine caused the most severe staining, followed by tea and coffee. After four weeks of immersion, brushing reduced surface staining by wine. On the contrary, chlorhexidine increased the staining effect of tea and coffee (p<0.05) when compared to the control specimens. In conclusion, we showed that common drinks stained the dental composite, but each by a specific mechanism that depended on external conditions such as the presence of chlorhexidine. PMID- 16706308 TI - Hardness development of dual-cured resin cements through different thicknesses of ceramics. AB - This study investigated the Knoop hardness of a thin layer in three dual-cured resin cements (Linkmax HV, Nexus 2, and Variolink II HV) irradiated through or not through different thicknesses (1 through 5 mm) of a machinable ceramic. Hardness was recorded at a series of time intervals up to five days, starting from the end of a light irradiation period. Increase in hardness was more rapid over the first 0.5 hour; thereafter it continued at a low rate until maximum hardness was attained. Ceramic thickness had a significant influence on hardness in all dual-cured resin cements, especially when ceramic thickness was more than 4 mm. In addition, it was noted that the polymerization of Nexus 2 seemed to be more dependent on light exposure compared with the other two materials. Variolink II HV and Linkmax HV, on the other hand, seemed to indicate the potential of being compensated by chemical curing to some degree. PMID- 16706309 TI - Development of beta-tricalcium phosphate/collagen sponge composite for bone regeneration. AB - Synthetic biomaterials have been developed and used for bone grafting. Here, we developed a biodegradable sponge composite for bone tissue engineering by combining beta-tricalcium phosphate (beta-TCP) and collagen. In addition, we sought to determine the optimal beta-TCP granules/collagen ratio by evaluating and bone formation in vivo. Porous beta-TCP granules were mixed with atelocollagen hydrochloride solution at various ratios--0.02, 0.05, 0.1, and 0.2 g/mL. The resultant mixtures were freeze-dried and subjected to dehydrothermal treatment in vacuo. The final composites obtained were designated beta TCP/collagen sponge composites (beta-TCP/CS). Through compression testing, it was found that the stress values for beta-TCP/CS (0.2 g/mL) were higher than those of the other three composites over the whole strain range. Histological evaluation at four weeks after implantation revealed that the collagen sponge had degraded and newly formed bone was present on the surface of the beta-TCP granules. At 12 weeks, the beta-TCP granules were completely degraded and remodeling of the lamellar bone was observed. PMID- 16706310 TI - Stress analysis of endodontically treated anterior teeth restored with different types of post material. AB - Finite element analysis was performed to evaluate stress distribution in maxillary central incisors treated endodontically and restored with a post and an all-ceramic crown. Tensile stress at tooth root was analyzed using two dimensional finite element models with different post diameters and lengths. One post length was 1/3 of the root (short), while the other was 2/3 of the root (long); one post diameter was 1/3 of the root (narrow), while the other was 2/3 of the root (wide). The following combinations were used for posts and cores: gold alloy cast post and core, commercial stainless steel post and resin core, and fiber post and resin core. Results showed that the fiber post produced less stress on the root dentin around the post tip than did the metal posts. This finding thus suggested that to reduce the stresses that cause root fracture, a long, thin fiber post should be used. PMID- 16706311 TI - Corrosion resistance and surface characterization of electrolyzed Ti-Ni alloy. AB - Ti-Ni alloy has been increasingly applied to medical and dental devices, such as coronary stents and orthodontic wires. This alloy contains nickel, which is known to give rise to cytotoxicity, metal allergy, and carcinogenicity. Therefore, the purpose of this study was to improve the corrosion resistance of Ti-Ni alloy by electrolytic treatment, whereby investigation was carried out using different acidic electrolyte compositions. As a result, specimens electrolyzed with lactic acid, water, and glycerol were found to show higher corrosion potential and release lower amount of titanium and nickel ions than mechanical-polished specimens (p<0.05). With the electrolytic treatment, nickel concentration in the surface oxide layer of Ti-Ni alloy decreased, and the thickness of the surface oxide layer increased. Based on the results of this study, it was shown that electrolytic treatment with suitable electrolyte could improve the corrosion resistance of Ti-Ni alloy, which is effective to produce medical and dental devices that utilize shape memory effect or superelasticity with better biocompatibility. PMID- 16706312 TI - Influence of polishing of denture base resin and metal surfaces on wettability with water and saliva. AB - The purpose of this study was to investigate the influence of the surface roughness of denture base resin and metal on wettability with water and saliva. Solid specimens were produced using heat-curing denture base resin and Co-Cr alloy. After polymerizing or casting these materials, specimens with four different types of surface roughness were produced by a progressive polishing process using polishing papers (#240, #400, and #1,000) and cotton buffs. Surface roughness decreased significantly as the polishing process progressed (p<0.0001). Contact angle increased as surface roughness in metal specimens decreased, but decreased as surface roughness in resin specimens decreased (p<0.0001). Furthermore, differences in contact angle between tap water and saliva were larger in metal specimens (p<0.0001). These results suggested that with regard to the tissue surface of a denture base, the influence of its roughness on adhesive force differed according to the type of denture base material. PMID- 16706313 TI - Study of the colors and retention of over-the-counter cosmetic veneers. AB - The purpose of this study was to observe the tooth colors and retention of a cosmetic veneer Fiche Pearl On containing shellac and five experimental formulations containing benzoin gum and ethyl cellulose. Tooth color was measured with a spectrophotometer before and after application of the veneers to the buccal or labial surface of experimental teeth. Through both in vitro and in vivo studies, the retention rates of veneers were assessed. In terms of color difference (deltaE*ab) between before and after veneer application, it ranged from 4.3 (Fiche Pearl On) to 15.0 (experimental formulation White Plus). In terms of retention rate, in vitro test found the retention rate of experimental formulation White Plus was significantly higher than that of Fiche Pearl On. In in vivo test where subjects were permitted to drink water and tea, it was found that the retention rate after two hours ranged from 94.0 to 98.5%. In conclusion, teeth applied with the experimental cosmetic veneer formulations showed better color and retention than teeth with Fiche. PMID- 16706314 TI - Effects of disinfecting alginate impressions on the scratch hardness of stone models. AB - This study investigated the effects of disinfecting alginate impressions on the scratch depth of resultant stone models. Eleven brands of alginate impression material and two disinfectants, 1% sodium hypochlorite and 2% glutaraldehyde, were used. Impressions were immersed in disinfectant solutions or stored in sealed bags after spraying with disinfectants, and then poured with a type V dental stone. The scratch depth of the stone model obtained from disinfected impression was measured. The storage of alginate impressions after spraying with disinfectants did not increase the scratch depth of resultant stone models. However, the effect of immersion in disinfectants on scratch depth varied with the brand of the alginate impression material. PMID- 16706315 TI - Study on the fracture strength of root reconstructed with post and core: alveolar bone resorbed case. AB - This study evaluated the influence of alveolar bone level on the fracture resistance of root restored with post and core. Forty-eight extracted human mandibular premolars were divided into six groups. Cast posts and cores were cemented (MN8 and MP8) or resin cores were built up with fiber posts and composite resin (FN8, FP8, FN4, and FP4). Post length was 8 mm (MN8, MP8, FN8, and FP8) or 4 mm (FN4 and FP4). Specimens were embedded 2 mm (MN8, FN8 and FN4) or 5 mm (MP8, FP8 and FP4) below cement-enamel junction. All specimens were loaded at 45 degrees to the long axis until fracture. With normal bone model, cast post and core (MN8) showed the highest fracture resistance (2262.4 N). However, in the resorbed bone model, there were no significant differences in fracture resistance between cast post and core and fiber post with composite resin. PMID- 16706316 TI - Effects of occlusal and brushing forces on wear of composite resins. AB - Effects of occlusal and brushing forces on the wear of composite resins were investigated using three different wear tests: simulated occlusal wear test, toothbrush wear test, and combined wear test which carried out toothbrush wear test and occlusal wear test alternately. Test specimens were prepared from four commercial composite resins. Worn volume and maximum worn depth were measured under different occlusal forces (40 N and 80 N) and brushing forces (1.5 N and 3 N) in the three wear tests. Worn surfaces were observed using a SEM. In all the three wear tests, both higher occlusal and brushing forces resulted in significantly greater worn volume and higher maximum worn depth. The effects of occlusal force on worn volume and maximum worn depth varied with different composites, indicating that the four tested composites showed different wear behaviors under different occlusal forces and brushing forces. It was suggested that their different wear behaviors most probably stemmed from the differences in their filler systems. PMID- 16706317 TI - Evidence-based nursing strategies to prevent ventilator-acquired pneumonia. AB - Mechanically ventilated patients are at risk of developing ventilator-acquired pneumonia (VAP). Critical care nurses working with intubated patients need to adopt strategies to prevent VAP within their clinical practice. VAP results from microaspiration of colonized secretions (the colonization of the aerodigestive tract by gram-negative bacteria found in the nares, oropharynx, sinus cavities, and the bacteria on dental plaque). This article is an overview of the risk factors, pathophysiology, and current strategies needed to prevent the occurrence of VAP, with focus specifically on the importance of oral health and oral care. Critical care nurses should promote the development and implementation of evidence-based oral care protocols and treatment regimens into their clinical areas to reduce the incidence and prevalence of VAP. PMID- 16706319 TI - Communication of medication orders by telephone--"writing it right". AB - Communication is commonly cited as a contributing factor to adverse events causing patient harm (Baker et al., 2004). There are numerous ways and reasons that communication failures can occur, such as poor handwriting, transcription errors, lack of verification, lack of integration of information, and ineffective team functioning. Errors that can occur in critical care with the verbal communication and receipt of a telephone order will be highlighted. These examples, together with proposed strategies for improving telephone order safety, are intended to promote awareness and potential practice changes in the critical care environment. PMID- 16706318 TI - Physiologic effects of first-time sitting among male patients after coronary artery bypass graft surgery. AB - OBJECTIVE: To study the physiologic effects of sitting among post-operative coronary artery bypass graft (CABG) male patients. METHODS: A prospective, repeated measures non-experimental design was used. Power analysis was used to calculate sample size based on pilot data from 19 subjects. Fifty-five males over the age of 18 years and having first-time CABG surgery were recruited. HR, BP, and SaO2 data were collected from the Marquette bedside monitor. SvO2 was measured by a blood gas sample from the pulmonary artery catheter. Baseline measurements were obtained on all subjects while supine in bed. Measurements were repeated immediately on sitting, after five minutes, after resuming the supine position, and again after 10 minutes. RESULTS: A repeated-measures ANOVA showed a significant time effect for HR (p < 0.001), SBP (p < 0.001), DBP (p < 0.001), MAP (p < 0.001), and SvO2 (p < 0.001), but not for SaO2. CONCLUSIONS: When post operative CABG male patients sit on the side of the bed for the first time, they experience an increase in HR and BP. Sitting involves increased oxygen consumption as evidenced by the drop in SvO2. Most patients recover to their baseline levels within 10 minutes of returning to the supine position. Nurses must be cognizant that this routine intervention may not be innocuous. Close monitoring of patients is essential and, with some, a graduated approach to sitting should be considered. PMID- 16706320 TI - There is no greater health epidemic in West Virginia than tobacco addiction! PMID- 16706321 TI - ACE inhibitor use and mortality reduction: do controlled trial results equal clinical practice? AB - Randomized trials have shown that angiotensin converting enzyme inhibitors (ACEIs) reduce mortality and morbidity and improve symptoms in many patients with heart failure. However, recent data show that the rate of ACEI prescriptions in West Virginia Medicare beneficiaries diagnosed with heart failure is not increasing. Data from the charts of patients who were discharged from 44 acute care hospitals during 2000 and 2001 were obtained, and these data were matched with current beneficiary data to determine if and when the patient died subsequent to the hospitalization of record. We examined data from 5,144 patients with heart failure for whom we had information on ACEI use, comorbidities, and contraindications, in addition to basic demographics. Patients who received angiotensin receptor blockers (ARBs) were excluded. Of these patients, 863 were eligible for ACEls, and 716 (83%) were discharged on an ACEI. Logistic regression showed that being discharged on an ACEI had a significant negative association with mortality one year later (P = .0009), reducing mortality in patients with heart failure by about one third. PMID- 16706322 TI - The adult with congenital heart disease. AB - There are approximately 800,000 adult survivors of congenital heart disease in the U.S., and this number continues to increase on an annual basis. It was only 50 years ago that the first intracardiac repair of a congenital heart defect was performed. Survival into adulthood has only been realistic in the last three decades. This has created a new patient population with a number of challenges such as the impact of adult onset diseases on their underlying cardiac pathology, cognitive, physical, and psychosocial development, as well as issues related to reproduction, employment and health insurance. Possibly the most significant obstacle is identifying healthcare professionals who can provide long-term care. This manuscript reviews the challenges and recommendations for the care of these patients. PMID- 16706323 TI - Maternal diabetes and perinatal outcomes in West Virginia Medicaid enrollees. AB - Diabetes Mellitus (DM) is a serious problem in West Virginia. Diabetes during pregnancy has been associated with several negative outcomes. This study tested the hypotheses that women who had DM during pregnancy would have more negative perinatal outcomes than those who did not and women with pre-gestational DM would have more negative perinatal outcomes than women with gestational DM. The population included 14,583 mothers enrolled in WV Medicaid in 2001 and 2002. The population was divided according to the presence of DM in the mother, and then sub-divided into pregestational and gestational DM groups. Results showed significant associations between maternal diabetes and occurrence of macrosomia, pre-term delivery and less than optimal one-minute Apgar scores. The pre gestational DM group had significantly more occurrences of macrosomia, pre-term birth, less than optimal Apgar scores, occurrence of complications of labor and/or delivery and occurrence of congenital anomalies than the gestational DM group. PMID- 16706324 TI - Bacillary angiomatosis following the use of long-term methotrexate therapy: a case report. AB - Bacillary angiomatosis, a rare and possibly fatal disease, occurs mainly in HIV infected patients. However, it has been reported in patients with cancer and in recipients of solid organs receiving immunosuppressive drugs such as cyclosporine. This case report describes a 66-year-old man who came to dermatology practice in Clarksburg for an initial visit. He had a longstanding history of psoriasis and psoriatric arthritis treated with methotrexate. A biopsy of a skin lesion on his neck confirmed the diagnosis of bacillary angiomatosis. PMID- 16706325 TI - Governor signs Omnibus Healthcare Bill (HB 4021). PMID- 16706326 TI - Methadone treatment preserving, saving lives. PMID- 16706327 TI - [Should COPD patients be ventilated?]. PMID- 16706328 TI - [Degree of public awareness regarding intensive care units (ICUs) and intensive care physicians in Castilla y Leon]. AB - AIM: To determine degree of public awareness regarding the activities and health care professionals that comprise intensive care units (ICUs) in the autonomous community of Castilla y Leon. DESIGN: Questionaire in the form of a true-false test dealing with a) description of an ICU; b) description of ICU patients, and c) degrees and qualifications held by ICU physicians. LOCATION: Waiting rooms of outpatient clinics and ICUs of 9 hospitals in Castilla y Leon. SUBJECTS AND METHODS: During the period from 1 October 2003 to 29 February 2004, there were surveyed a group of those persons accompanying outpatients arriving for appointments (OP, n = 2,293), and a group comprised of relatives of ICU patients (ICU, n = 727) upon discharge from the ICU. RESULTS: The average age of those subjects surveyed was 45.6 years old. 62.1% were women and 52.8% had received education through secondary level or higher. Of the 1,354 analyzed OP questionnaires, 27.7%, 25.5% and 48.4% responded correctly to questions A, B and C, respectively. Of the 284 analyzed ICU questionnaires, 38.6%, 41.0%, and 63.5% responded correctly to questions A, B and C, respectively. Differences between results from the two surveyed groups were statistically significant, and the best results from the ICU group were found among those subjects whose family members had remained in ICU for 2 days or more. CONCLUSIONS: In the described scope, of the citizens of Castilla y Leon, 27.7% know as it is a ICU, 25.5% know what type of patients usually is entered there, and 48.4% recognize their doctors like ICU specialists specifically. These knowledge improve significantly after having some relative entered in ICU for more than 2 days. PMID- 16706329 TI - [Prognosis for acute exacerbation of chronic obstructive pulmonary disease in mechanically ventilated patients]. AB - OBJECTIVE: To determine the variables associated with prognosis for acute exacerbation of chronic obstructive pulmonary disease (AECOPD) in mechanically ventilated patients. DESIGN: Prospective cohort study with retrospective analysis. LOCATION: 361 Intensive Care Units (ICU) in 20 countries. PATIENTS AND METHODS: There were included in the study 522 patients who required mechanical ventilation for more than 12 hours due to an acute exacerbation of chronic obstructive pulmonary disease (AECOPD). In order to determine those variables associated with mortality, there was performed a recursive partition analysis in which the following variables were included: demographics, arterial blood gas prior to intubation, complications arising during mechanical ventilation (barotrauma, acute respiratory distress syndrome, ventilator-associated pneumonia, sepsis), organ dysfunction (cardiovascular, renal, liver, coagulation) and duration of ventilatory support. INTERVENTIONS: None. VARIABLES OF PRIME IMPORTANCE: ICU mortality. RESULTS: ICU and hospital mortality rates were 22% and 30%, respectively. Variables associated with mortality were cardiovascular dysfunction, renal dysfunction and duration of ventilatory support > 18 days. Median durations were as follows: mechanical ventilatory support, 4 days (P25: 2, P75: 6); weaning from ventilatory support, 2 days (P25: 1, P75: 5); stay in intensive care unit, 8 days (P25: 5, P75: 13); stay in hospital, 17 days (P25: 10, P75: 27). CONCLUSIONS: Mortality in the studied cohort of patients with AECOPD was associated with cardiovascular dysfunction, renal dysfunction and prolonged mechanical support. PMID- 16706330 TI - [Thrombolysis in cardiac arrest]. AB - Both acute myocardial infarction and pulmonary thromboembolism are responsible for a great number of cardiac arrests. Both present high rates of mortality. Thrombolysis has proved to be an effective treatment for acute myocardial infarction and pulmonary thromboembolism with shock. It would be worth considering whether thrombolysis could be effective and safe during or after cardiopulmonary resuscitation (CPR). Unfortunately, too few clinical studies presenting sufficient scientific data exist in order to respond adequately to this question. However, most studies they show that thrombolysis applied during and after CPR is a therapeutic option that is not associated with greater risk of serious hemorrhaging and could possibly have beneficial effects. On the other hand, experimental data exists which show that thrombolytics can attenuate neurological damage produced after CPR. Nevertheless, clinical trials would be necessary in order to adequately establish the effectiveness and safety of thrombolysis in patients who require CPR. PMID- 16706331 TI - [Code of ethics of the Spanish Society of Intensive Care, Critical and Coronary Units (SEMICYUC)]. PMID- 16706332 TI - [Acute coronary syndrome as presently categorized]. PMID- 16706333 TI - [Early onset of torsades de Pointes and elevated levels of serum troponin I due to acute arsenic poisoning]. AB - Most cases of acute arsenic poisoning occur through accidental or voluntary ingestion of pesticides or insecticides, and all body systems are affected. Arsenic can prolong the QT interval and lead to torsades of Pointes, a crucial type of arrhythmia characteristic of such QT interval prolongation. In our revision of the literature, there have been found only 5 cases of torsades of Pointes due to acute arsenic poisoning. Recently, there have been published four additional cases in patients with refractory or recurrent acute promyelocytic leukemia being treated with arsenic trioxide. In all nine cases, torsades of pointes appeared slowly after poisoning. Herein is described a case of acute arsenic poisoning which led to an early onset of torsades of Pointes, hypopotasemia and high levels of serum troponin I. PMID- 16706334 TI - [Multi-organ failure secondary to acute fatty liver of pregnancy]. PMID- 16706335 TI - [Pneumothorax, pneumoperitoneum and retropneumoperitoneum due to spontaneous colon perforation]. PMID- 16706336 TI - [Cardiac herniation thru pericardial window]. PMID- 16706337 TI - [Analysis of rat 24h heart rate variability based on telemetry technology]. AB - In this study ECG signal of unstrained rat was recorded by telemetry device, and heart rate variability (HRV) was analyzed in order to evaluate 24h autonomic nervous activity. The results demonstrated an obvious circadian rhythm in the autonomic nervous activity: sympathetic activity being dominant during wake phase, and parasympathetic activity, dominant during sleep phase. The ratio of the low frequency to high frequency components in HRV power spectrum (LF/HF) fluctuates with the change in the sleep stages. It is concluded that 24h HRV analyses may reveal plentiful information about the behavior of autonomic nervous system and thus facilitate the investigation of its regulating role in physiological and pathological processes. PMID- 16706338 TI - [A data interface based on USB bus technology for full auto patch-clamp system]. AB - A USB bus based data interface technology for full auto Patch-Clamp system is discussed in the article. The main controller is CY2131QC (Cypress) and the logic controller is EPM3256A (Altera). Optocouplers are used to get rid of the noise from the interface. It makes the installation of the Patch-Clamp system easier by using the USB bus, and is suitable for the new generation of the Patch-Clamp system with a high speed of 1M bytes/s. PMID- 16706339 TI - [Stochastic nonlinear evolutional model of the large-scaled neuronal population and dynamic neural coding subject to stimulation]. AB - In this paper, we investigate how the population of neuronal oscillators deals with information, and analyze the dynamic evolution of neural coding when the outer stimulation acts on it on the base of our former work. By numerically computing for the model, we obtain the figure of average number density, which is used to describe the action potential of the neurons within population in three dimensional space, namely the dynamic evolution of neural coding. The result of numerical analysis indicates that the model in this paper can be used to describe the evolutional process of abundant mutual interactional neurons acted by outer stimulation. The numerical result also proves that only the suitable stimulation can change the coupling structure of neurons. And the evolution model given in this paper incarnates the neural plasticity. PMID- 16706340 TI - [The study and implementation of DICOM conformance in the network printing of PACS]. AB - To implement the network printing in PACS (Picture Archiving and Communication System), each level of DICOM (Digital Imaging and Communication in Medicine) print management service is defined by means of object oriented method, and the corresponding events and properties are implemented with C++ language. It is tested on laser printers of AGFA, KODAK and FUJI to determine whether the network environment supports the print management service and implements the presentation LUT (Look Up Table) correctly. The quality control of printing at the level of data flow is also assessed. The program finally succeeds in realizing the network printing based on DICOM in hospital environment. PMID- 16706341 TI - [Single-trial estimation of visual evoked potentials in single channel single trial estimation]. AB - We constructed a Brain-computer interface-based mental speller which realizes user-computer interaction. The feature signals of user's intention are embedded in spontaneous EEG background. Single-trial feature estimation should be used on this online occasion instead of the grand average usually used in cognitive or clinical experiments. To demonstrate this technique beyond laboratories, fewer EEG recording channels are preferred. A unique paradigm, which is called imitating-natural-reading, was exploited to induce visual evoked potentials. We explored the single-trial estimation of VEP recorded in single channel using support vector machine on three subjects, and obtained satisfactory data, the classification accuracy being 92.1%, 94.1% and 91.5%, respectively. These results put forward a significant step fowards the ultimate realization of our mental speller. PMID- 16706342 TI - [R-wave detection of ECG signal by using wavelet transform]. AB - The detection of R-wave of ECG is essential to the analysis of the heart rate variability (HRV). In this paper, an R-wave detection method using wavelet transform(WT) is presented in line with the principle of discrete wavelet transform(DWT) and multi-resolution technique (MRT). We made use of the special properties of dbl wavelet in time-domain, decomposed the original ECG signals into 3-level detailed signals on different frequency bands by using DWT with Mallat algorithm, and got appropriate threshold values in different high frequency bands to distinguish R-wave. It is concluded that the algorithm had significant effects on it, which is verified by MIT/BIH (Massachusetts Institute of Technology/Boston's Beth Israel Hospital) ECG Database. The results show that R-wave could be detected accurately and localized precisely by this method, even when the patient was seriously sick or the signal was disturbed by noise. Consequently the method has a quite high locating precision (its error is not more than two sampled points and about 85 percent of the points of R-wave in ECG signal are localized precisely) and the correct detection rate of R-wave is 99.8% by using wavelet transform, so this method is quite feasible. PMID- 16706343 TI - [Research on EEG classification with evolving cascade neural networks]. AB - To correctly classify EEG with different mental tasks, a new learning algorithm for Evolving Cascade Neural Networks (ECNNs) is described to avoid over-fitting of a neural network due to noise and redundant features. The learning algorithm calculates the value of a fitness function on validate set and accordingly updates the connection weights on training set. The learning algorithm uses the regularity criterion for selecting the neurons with relevant connection. If the value Cr calculated for the rth neuron is less than the value Cr-1 calculated for the previous (r-1) neuron, the features that feed the rth neuron are relevant, else they are irrelevant. An ECNN starts to learn with one input node and then, adding new inputs as well as new hidden neurons, evolves it. The trained ECNN has a nearly minimal number of input and hidden neurons as well as connections. The algorithm is applied to classify EEG with two mental tasks. The trained ECNN has correctly classified 83.1% of the testing segments. It shows a better result, compared with a standard BP network. PMID- 16706344 TI - [The research and development of new diagnostic gene chips for SARS coronavirus]. AB - The loci of cDNA sequences for valid diagnosis have been identified through the selection of the genome of SARS coronaries. The gene chips for diagnosing such virus have been developed, based on our own-developed technology for manufacturing and application of gene chips. The diagnoses given by such gene chips were consistent well with the reports of clinical laboratories (94.29%) and the sensitivity reached 10(-2)/ml virus molecules. This method is well suited for the clinical use in SARS coronaries diagnosis. PMID- 16706345 TI - [Evaluation of back muscle function based on EMG time-frequency spectrogram analysis]. AB - EMG has been extensively used to study function of back muscle, which plays an important role in the objective assessment of occupational low back pain, yet the inherent large variability of EMG signals across subjects produced by reasons already known or unknown may mask true biological differences. Some useful parameters abstracted from 8-channel EMG signal in time-frequency domain are used or proposed and calculated in this paper to decrease this variability, when comparing the possible difference between low back pain patients and normal control group. The calculation of instantaneous median frequency is improved to decrease the interruption of background noise, and different power densities in EMG time-frequency spectrogram are observed in two tested groups. PMID- 16706346 TI - [A method of iris image quality evaluation]. AB - Iris image quality evaluation plays a very important part in iris computer recognition. An iris image quality evaluation method was introduced into this study to distinguish good image from bad image caused by pupil distortion, blurred boundary, two circles appearing not concentric, and severe occlusion by eyelids and eyelashes. The tests based on this method gave good results. PMID- 16706347 TI - [A method based on image processing and analyzing technology for estimating the activity of mesenchymal stem cells]. AB - Cell culture is one of the usual methods for studying living cell and tissue, the method presented in this paper is based on image processing and analyzing technology for activity estimation of mesenchymal stem cells (MSCs). The existing activity estimation methods are costly, complex and invasive. In this method, thresholding is used to preprocess image and to separate out the growth hallow. Then the area is calculated by counting the pixels of the growth hallow. The changes of the activity estimated by this method are similar to those by corresponding cellular experiments. Compared with the existing methods in biology, medicine or medical cellular science, this method is easier, faster, cost-effective and non-invasive. The proposed method has been proved to be efficient by primary experiments of MSCs. PMID- 16706348 TI - [Study of color blood image segmentation based on two-stage-improved FCM algorithm]. AB - This paper introduces a new method for color blood cell image segmentation based on FCM algorithm. By transforming the original blood microscopic image to indexed image, and by doing the colormap, a fuzzy apparoach to obviating the direct clustering of image pixel values, the quantity of data processing and analysis is enormously compressed. In accordance to the inherent features of color blood cell image, the segmentation process is divided into two stages. (1)confirming the number of clusters and initial cluster centers; (2) altering the distance measuring method by the distance weighting matrix in order to improve the clustering veracity. In this way, the problem of difficult convergence of FCM algorithm is solved, the iteration time of iterative convergence is reduced, the execution time of algarithm is decreased, and the correct segmentation of the components of color blood cell image is implemented. PMID- 16706349 TI - [Semantic extension to DICOM standard]. AB - The Digital Imaging and Communications in Medicine (DICOM) Standard was developed by National Electrical Manufacturers Association (NEMA) and the American College of Radiology (ACR) for medical image archiving and communication. The Standard is widely used in medicine now. But it does not define image semantic content sufficiently, so there is a difficulty in the implementation based on semantic content. In this article are discussed the semantic hierarchy structure in medical image and the method to describe the image content used in the DICOM Standard. We point out the limitations of DICOM in image semantic content description, and then set forth a way to extend the semantic content description by using DICOM private attributes. PMID- 16706350 TI - [An experimental study on mechanical properties of fiber layers in anulus fibrosus of lumbar interverbral disc 4,5]. AB - We tested the mechanical properties of fiber layers in adult anulus fibrosus of lumbar interverbral disc 4,5. Aulus fibrosus of lumbar interverbral disc was delaminated by using microsurgical technique. 8 testing points were taken in each layer according to the fibers going and 5 testing specimens were taken from each testing point. The length was 15-20mm, the width 1-2mm, and the thickness 0. 1-0. 5mm. By using tension test,the relationship curves of stress and strain were tested, the fitting curves and equations were obtained, and then the elastic modulus, damage strain and damage stress were obtained. Therefore we knew the parameters and equations of mechanical properties of each testing point. We got the result that the elastic modulus of each testing point, along the radius from outside to inside, decreased with the increase of layers. The damage stress decreased linearly from outside to inside. The damage strains of the outside 9 layers increased slightly with the increase of layers, and the others were almost constants, i. e. 0.34 +/- 0.14. We get the conclusion that analus fibrosus of intervertebral disc has special mechanical properties corresponding with its functions, which is in a close relationship with lumbar vertebral diseases. PMID- 16706351 TI - [Comparisons of hemodynamics in 1-way and 2-way CABG]. AB - For the purpose of improving the hemodynamics of CABG, the authors presented a new amelioration measurement of symmetric 2-way bypass graft. The physiological blood flow in 1-way and 2-way bypass graft of coronary is simulated with finite element method, and the data on computational hemodynamics of these two cases were compared. The temporal-spatial distributions of hemodynamics during the cardiac cycle such as flow patterns and wall shear stress in the vicinity of anastomosis were analyzed. The study results showed that 2-way bypass graft was of more rational hemodynamics when compared with 1-way bypass graft, and it could improve the flow conditions and decrease the probability of restenosis. PMID- 16706352 TI - [Model experiment and numerical simulation of marginal pressure in the flow field downstream of the stenostic vessel in vitro]. AB - For the defects of interpreting the relationship between the distribution and severity of vascular stenostic disease and the low shear stress, the study focus on and explore the rules of the pressure distribution and variation downstream of the stenostic vessel with the numerical simulation and the hydrodynamic model of turbulent flow separation area for in vitro experiment. The results reveal that there is a good compatibility between the model experiment and the numerical simulation; there exists a marked pressure decrease downstream of the stenosis. And the distribution area of low pressure is similar to the characteristic distribution of the stenostic vessel diseases. The study suggested that the numerical simulation can be applied in the study of micro-flow field; the presure distribution and variatioin downstream of the stenostic vessel maybe another factor to trigger the pathogenesis of vessel stenostic diseases. PMID- 16706353 TI - [Cellular automata approach to biological pattern formation (I): the aggregation pattern in dictyostelium discoideum]. AB - The investigation of the mechanism of biological pattern has been an important topic of life sciences, especially, of developmental biology, for a long time. It is an interdisciplinary problem and many researching data have been obtained and some theories have been structured from many points of view in science. However, up to now, the actual mechanism is still a fascinating puzzle and needs more studies. In this paper, we try to construct a cellular automata model of biological pattern. This model defines the individual model cells and their behaviors, cell-cell interactions, and cell-environment interactions. As an application, we present a new discrete model to simulate the aggregation phase of the development of Dictyostelium discoideum with the concept of "inducing switch". PMID- 16706354 TI - [Observation on the characters of infrared radiation spectrum of acupoints in normal humans and CHD patients]. AB - In this experiment the data of the infrared radiation spectrum of acupoints in human body were obtained through an ingenious device. In thirty-one healthy volunteer subjects and thirty-one patients suffering from coronary heart disease, we detected the infrared radiation spectra from some acupoints and compared them with those of the healthy subjects. The curves of patients showed more differences. After analyzing the data, we noted statistically significant differences in some fields between the healthy subjects and the patients. And after reducing the data, we observed significant differences in larger fields. So utilization of reduced data from patients for analysis is our choice. PMID- 16706355 TI - [Mechanical properties of rat HCC adhesion to collagen I and its relationship with cell cycle]. AB - The mechanical properties of tumor cells adhering to extracellular matrix (ECM) are closely related with their invasion and metastesis. In this study we investigated the adhesive mechanical properties between hepatocellular carcinoma cells(HCC) and the collagen I coated surfaces from the viewpoint of cell cycle by coupling cellular biology and cellular mechanics, using micropipette aspiration and cell synchronization technique. The results showed that the synchronous G1 and S phase HCC cells were achieved by use of thymine-2-desoryriboside, colchicines sequential blockage method and double thymine-2-desoryriboside blockage method, and that the synchronous rates of G1 and S phase HCC amounted to 74.09% and 90.39% respectively. Within the ranges of dosing and timing in this study, the adhesion of HCC cells to collagen I displayed dose dependent and time dependent patterns. S phase cells had small force of adhesion to collagen I as compared with G1 phase and controlled cells(P<0.001), which suggested that G1 phase HCC may play an important role in the step of invading interstitial connective tissue in the metastasis pathway of HCC through blood circulation. These are of significance to unveiling the mechanism of HCC metastasis. PMID- 16706356 TI - [Quantification study of bone adaptive model based on experiment of rapid-growing rats in different stress environment]. AB - Presented is a method to set up a quantification model of bone growing and remodeling adaptation, which integrates animal experiments, parameter identification of mathematical functions and technique of computer simulation. By designing a new animal experiment, we investigate the effects of growing and remodeling of the rat femurs in different stress environments, gather the bone mineral density (BMD) of proximal femur in the same interval for the unknown parameter (B and K) inversion of bone growing and remodeling equation and create the femur three-dimensional geometrical model based on CT images. The model in this paper can not only numerically measure the relation between outer stimulus and the femur BMD variation of rapid growing rats, but also predict the growth trend of rat femur under different stress environments in its whole lifecycle. The thought and method of creating the model in this paper can be used for reference to modeling human bone growth and remodeling. PMID- 16706357 TI - [Research on dynamic and synchronous changes of microcirculatory ultrastructure and free radical in rat cerebrum after exhaustive swimming]. AB - This research is designed to detect the dynamic changes of microcirculatory ultrastructure and the MDA content, SOD and GSH-px activity in rat cerebrum immediately, 24h and 48h after exhaustive swimming. The results showed that there were obvious changes of ultrastructure in rat cerebral microcirculation immediately after exhaustive swimming. The changes were resumed in some degree after 24h and recovered almost to the normal after 48h. Immediately after exhaustive swimming, the MDA content did not change obviously; the activity of SOD increased markedly, and then it decreased markedly after 24h, but it increased again after 48h. The activity of GSH-px descended markedly after 24h; it descended synchronously with the decrease of SOD activity, showing that the consumption of SOD and GSH-px should be the most at this time. Therefore, the free radical should be removed in time for reducing the cerebral injury and promoting the recovery of cerebral microcirculation. PMID- 16706358 TI - [Influence of stretch strain and pressure on expression of osteoblasts' FOS protein]. AB - Uniaxial stretch strain and compressive pressure of 2 atm were applied to rat's osteoblasts, and then immunohistochemistrical staining was used to detect the expression of osteoblasts' c-fos gene. The experiment result indicated the osteoblasts' FOS proteins increased prominently, and the FOS proteins concentrated in nucleolus after having endured two different kinds of loadings. It is very important to prompt stress and strain in promoting osteoblast proliferation. PMID- 16706359 TI - [Influence of different convergence angles on the retention of cast metal post and core]. AB - Fifty-four simulated tooth roots made of polymethacrylate (PMMA) were divided into six groups according to the root canal convergence angles of 0 degrees, 3.93 degrees, 5.71 degrees, 7.48 degrees, 11.31 degrees, 14.71 degrees. Cast post and cores were manufactured and cemented with Zinc phosphate cement (ZP). The retention capacities of the posts were tested. The bonding strength of ZP at different convergence angles were tested with some casted wafers, and linear regression was made. So the effect of interlocking and the proportion could be calculated. With these results, we made some curvilinear regressions. The results showed that the retention of cast post declined and the bonding strength of ZP fell with the increase of convergence angles. There were significant differences in the bonding strength and retention among groups, P<0.05. The experiment indicates that the dentists should reduce the convergence angles of posts as much as possible to obtain greater retention. PMID- 16706360 TI - [A mechanical model of knee joint in sagittal plane]. AB - A sagittal plane model of knee joint based on crossed-four-bar-linkage-based tibiofemoral joint model has been developed using geometric and force equilibrium constraints. The model predicts and explains the movement of contact point on the patella and femur, variation of patellar and patellar tendon angle, variation of patellar mechanism angle and variation of patellofemoral contact force and patellar tendon force. The computed results agree well with the published experimental results. PMID- 16706361 TI - [Study on preparation of the pH sensitive hydroxyethyl chitin/poly (acrylic acid) hydrogel and its drug release property]. AB - Hydroxyethyl chitin (HECH) is a water soluble chitin derivative made by etherification of chitin, ethylene chlorohydrin was used as etherification reagent in this reaction. A novel interpenetrating polymer network (IPN) composed of HECH/PAA was prepared. The IR spectra confirmed that HECH/PAA was formed through chemical bond interaction. The sensitivity of this hydrogel to temperature and pH was studied. The swelling ratio of this hydrogel in artificial intestinal juice is much greater than that in artificial gastric juice. The IPN hydrogel exhibited a typical pH-sensitivity, and its degree of swelling ratio increased with the increase of temperature. The sustained-release drug system of Dichlofenac potassium was prepared by using HECH/PAA as the drug carrier. The release experiment showed a perfect release behavior in artificial intestinal juice. This IPN is expected to be used as a good drug delivery system of enteric medicine. PMID- 16706363 TI - [Synthetic and photo-immobilization of photo-active tumor necrosis factor-alpha]. AB - Tumor Necrosis Factor is one of the cell factors with much stronger anti-tumor activity. In this study, photoactive arylazide-4-azidobenzoic acid was modified to Tumor Necrosis Factor-alpha (TNF-alpha). The IR data (2127cm(-1)) was given to confirm the modification. By photo-immobilization, this modified TNF-a was immobilized on cell culture polystyrene. Both the relation between the amount of TNF-alpha in feed and immobilized TNF-alpha and the influence of different UV irradiation time on photo-immobilization are discussed. Microscopic observations of the photo-immobilization TNF-alpha were made by use of scanning electrom microscope and atomic force microscope. PMID- 16706362 TI - [The effect of surface free energy parameters of diamond-like carbon films deposited on medical polyethylene terephthalate on bacterial adhesion]. AB - Diamond-like carbon (DLC) films were deposited by acetylene plasma immersion ion implantation-deposition (PIII-D) on biomedical polyethylene terephthalate (PET). The capacities of Staphylococcus aureus (SA), Staphylococcus epidermidis (SE), Escherichia coli (EC), Pseudomonas aeruginosa (PA) and Candida albicans (CA) for adhesion to PETs are quantitatively determined by the plate counting and Gamma ray counting of 125I radio labeled bacteria in vitro. The results indicate that the capacities of five types of bacteria for adhesion to PETs are all suppressed by C2H2 PIII-D (P<0.05). The surface energy components of the various substrates and bacteria are calculated based on measurements in water, formamide and diiodomethane and Lifshitz-van del Waals/acid-base approach (LW-AB). The surface free energies obtained are used to calculate the interfacial free energies of adhesion (deltaF(adh)) of five kinds of bacteria on various substrates, and the results show that it is energetically unfavorable for bacterial adhesion to the DLC films already deposited on PET by C2H2 PIII-D. PMID- 16706364 TI - [Influence of biomateriai on focal adhesion kinase of platelet: ex vivo study]. AB - This study was aimed to detect the biomaterial-induced phosphorylation level of platelet focal adhesion kinase, a key component of signal transduction pathways. Flat tubings, made from 5 biomaterials: organotin-PVC, PU50, PU60, PU70, and PTFE,were connected to an ex vivo flow circuit with canine femoral artery and vein. After 5 min, the adherent platelets on the inner surface of the tubing were lysed, then FAK and phospho-FAK were examined, using immunoprecipitation and Western blot. The results showed a significant increase in the FAK value on the PVC and PTFE surface,compared to the PUs; among the PUs, the FAK value decreased in the following order: PU70/PU60/PU50. The phospho-FAK value on the five groups of surface decreased in the following order: PVC/PTFE/PU70, PU60, PU50; there was no significant difference among the PUs with regard to the phospho-FAK value. In conclusion, after platelet-material interaction, biomateriais may lead to the phosphorylation of FAK of platelets;the level of phosphorylation of FAK has been positively linked to the thrombogenicity of materials; the phospho-FAK value may be a valid molecular parameter of the haemocompatibility assessment; the FAK value may supply references to the haemocompatibility assessment. PMID- 16706365 TI - [The synthesis, characterization and in situ intestinal absorption of different molecular weight scutellarin-PEG conjugates]. AB - In this report, highly water soluble esters of scutellarin with different molecular weight polyethylene glycol (PEG) were synthesized in order to improve the bioavailability of scutellarin. The physicochemical properties, the stabilities under different conditions and the in situ intestinal absorption in rats of the conjugates were investigated. By PEG modification, the greatly increased water solubility and desirable partition coefficient of scutellarin were obtained. These compounds function as prodrugs i. e. breakdown occurred in a predictable fashion: the t1/2 of them in PBS buffer at pH7.4 was above 12 h (37 degrees C) in vitro, while in plasma a rapid breakdown was observed, with a t1/2 of about 1. 5-3 h. The stabilities of the prodrugs were improved according to the increase of the molecular weight of PEG, thus, PEGylated prodrugs with desirable rates of hydrolysis could be obtained by the use of variable molecular weight PEGs. The PEGylation could enhance the absorption of scutellarin in rat intestine, and the absorption of scutellarin and its PEG conjugates by intestine was mainly via passive transport, for when the concentration was raised, the uptake did not appear saturable, and the permeability coefficient kept at an equilibrium level. When the molecular weight of PEG increased from 200 to 1000 Da, the absorption of the conjugates decreased. In conclusion, by overall consideration of the yield, stability and absorption, the present authors estimate that the PEG molecular weight used for the PEGylation of scutellarin should be within the range of 400-1 000 Da. PMID- 16706366 TI - [Biological evaluation of artificial skin substitute]. AB - Multiple kinds of Artificial Skin Substitute are now available. However, except for the Homo Skin Graft there is no Artificial Skin Substitute that can be used as permanent Artificial Skin Substitute. During the past 20 years, more and more scholars around the world have expressed increased interests in the research and development of Artificial Skin Graft that can be utilized as satisfying permanent Artificial Skin Substitute. We conducted our research on the biological evaluation of medical devices of Collagen-Chitosan(C-C) Artificial Skin Substitute according to the National Standard (GB/T16886. 1-1997). The following experiments were conducted: (1)Cytotoxicity, (2)Systemic toxicity(acute toxicity), (3)Haemocompatibility, (4)Sensitization, (5)Intracutaneous reactivity, (6)Pyrogen test, (7)Genotoxicity. The experiment results demonstrate that all biological functional indexes of the Artificial Skin Graft meet the National Standards. Therefore, we conclude that C-C Artificial Skin Graft is characteristic of good biological compatibility. It is non-irritant and has no systemic and cellular toxicity, no genotoxicity, no pyrogen, and no allergen. PMID- 16706367 TI - [Synthesis and property of nano-fumed silica derivative with quaternary ammonium group]. AB - Nano-fumed silica derivative with quaternary ammonium group was synthesized and the antimicrobial activity was investigated. The nano-fumed silica derivative was investigated by Fourier-transform infrared spectroscopy (FTIR). The zeta potential and the size of the nano-fumed silica derivative were measured. The antimicrobial properties of the nano-fumed silica derivative against selected microorganisms were tested by the quantitative suspension method. The zeta potential showed that the isoelectric points of nano-fumed silica and modified nano-fumed silica are pH=4. 8 and pH =10.5-10.8, respectively, and the shift of the isoelectric point is due to the quaternary ammonium salt. The obtained nano fumed silica derivative inhibited the growth of Gram-negative (Escherichia coli), Gram-positive bacteria (Staphylococcus aureus), and fungus (Candida albicans). The inhibiting effect of nano-fumed silica derivative on the microorganisms varied with the time of exposure. The bacteriostatic rates were noted to be 99.99%, 99.99% and 94.12% after 15 minutes' exposure, respectively. Thus the results indicate that nano-fumed silica derivative with quaternary ammonium group has significant inhibitory effect on the growth of bacteria. PMID- 16706368 TI - [An experimental research on different temperature sintered bone as carrier of bone morphogenetic protein]. AB - This study was conducted to find perfect temperature sintered bone as carrier of bone morphogenetic protein (BMP). The different temperature active sintered bones, which were made up of calcine bone and bone morphogenetic protein, were implanted into the defects of rabbit radius. Compared with the sintered bone of 600 degrees C, the sintered bone of 900 degrees C and 1200 degrees C could induce more pieces of bone formation and be replaced by new bone. There were more pieces of new bone formation in sintered bone of 900 degrees C and 1200 degrees C than those in sintered bone of 600 degrees C (P<0.05). There was no difference between the sintered bone of 900 degrees C and 1 200 degrees C (P>0.05). In comparison with the sintered bone of 600 degrees C and 1200 degrees C, the sintered bone of 900 degrees C may be the choicest carrier of bone morphogenetic protein. PMID- 16706369 TI - [Grafting and characterization of poly (ethylene glycol) on polysulfone sheets]. AB - Grafting of poly (ethylene glycol) (PEG) on the surface of polysulfone (PSF) sheets by simultaneous or sequential UV irradiation with 4-azidobenzoic acid as the photocoupler was carried out. Water contact angle measurements showed that there was a great improvement of hydrophilicity on the grafted surface. X-ray photoelectron spectroscopy suggested that the area covered by PEG be 77.3% and 41.9% respectively after grafting by simultaneous and sequential pathways. With atomic force microscope (AFM), obvious difference in the shape and the phase mode was observed between surfaces of PEG-g-PSF sheets made by these two pathways. Evidences implied that simultaneous pathway would produce a branched PEG layer on the surface, while sequential pathway was coupled with a "pan-cake" PEG layer on it. This study provides the foundation for further advancement in tethering brush like PEG on PSF hollow fiber membranes. PMID- 16706370 TI - [A study on the mechanical properties of poly-D,L-lactic acid/hydroxyapatite compound fibers]. AB - The PDLLA/HA compound fibers with excellent properties could be obtained by melt spinning. This study inquired about the mechanical properties, change rule and influencing factors. The results showed that the composites fibers of mechanical properties were good in the PDLLA(molecular weight 120,000) to which were added the 4-20 microm HA particles. In the matrix of PDLLA with accretion (10%) of HA particles, the tensile strength of the compound fibers was the highest, compared with the others. The tensile strength of the compound fibers of PDLLA (molecular weight 20,000-300,000) was high. The tensile strength of the compound fibers decreased with the increase of the fiber diameter, and the PDLLA/HA fibers 40-60 micrometers in diameter had the highest elongation at break. PMID- 16706371 TI - [Cloning of major outer membrane protein gene of Legionella pneumophila and detection of its expression in prokaryotic cell]. AB - In this study, the ompS gene, a major outer membrane protein gene of Legionella pneumophila, was obtained from the DNA of Legionella pneumophila by PCR. The gene was cloned into prokaryotic expressional plasmid pUC18 to construct recombinant plasmid. The recombinant plasmid was transformed into E. coli strain BL21. The identification was made by means of restriction enzyme analysis, polymerase chain reaction, DNA sequencing analysis, SDS--polyacrylamine gel electrophoresis analysis and Western blot. The results showed that the ompS gene of 914 bp was amplified from Legionella pneumophila DNA, the recombinant plasmid pLPompS was constructed and its expression in prokaryotic cell was detected successfully. PMID- 16706372 TI - [Screening for endocrine exophthalmos-associated genes/cDNA fragments]. AB - This study was conducted to screen the endocrine exophthalmos-associated genes or cDNA fragments and provide a basis for exploring the pathogenesis. The cDNA from the thyroid tissues of patients with hyperthyroidism and endocrine exophthalmos (HT&EE) was used as tester cDNA, and that from the thyroid tissues of patients with HT but free from EE was used as driver cDNA. The subtractive PCR products between tester and driver cDNA were obtained through two cycles of subtraction hybridization and two cycles of PCR by using suppression subtractive hybridization, and then were inserted into pT-Adv, a cloning vector. The ligated DNAs were transformed into E. coli DH5alpha competent cells and incubated for proper blue/white color development. Forty-eight white colonies were randomly picked and their inserts were colony PCR amplified. The PCR product from one of the colonies contained two inserts; the others contained single insert, having a size of 0.2 kb to 2 kb. The inserts of transformants were arrayed on nylon membrane. After cDNA/Rsa I digestion the thyroid tissues of patients with HT, and of patients with HT&EE, were labeled with digoxigenin; the nylon membranes were then hybridized respectively with the two cDNA probes for a high throughput screening for positive clones. The clones which were hybridized with the cDNA probe of HE&EE patients but not hybridized with the probe of the HT patients or showed only faint signal of hybridization, were chosen as positive clones and their inserts were candidates for the endocrine exophthalmos genes or cDNA fragments. About 50% of the clones were confirmed as positive clones. The cDNA fragments in the positive clones were the endocrine exophthalmos-associated genes or cDNA fragments. Endocrine exophthalmos PMID- 16706373 TI - [Evaluation of three in-vitro refolding methods for human-derived anti-CTLA4 scFv expressed in E. coli]. AB - To evaluate the efficiency of three in vitro refolding methods for a humanized single-chain Fv antibody against human CTLA4(CD152) expressed in E. coli, the denatured and purified inclusion bodies (IBS) were refolded by dilution, dialysis and in situ refolding via Immobilized Metal-Ion-Affinity Chromatography (IMAC), respectively. The concentration of refolded scFvs was examined by Bradford method. And the antigen binding activity of the refolded scFvs was analyzed by indirect cell-ELISA. The highest and lowest refolding yields could be obtained by dialysis and in situ refolding via IMAC, respectively. The binding activity of the refolded scFv by dialysis was 1.95-fold higher than that by dilution, 4.13 fold higher than that by in situ refolding via IMAC (GSH/GSSH excluded) and 3.63 fold higher than that by in situ refolding via IMAC (GSH/GSSH included), respectively. In conclusion, a high refolding yield and binding activity of scFv with natural conformation could be obtained by dialysis in the condition of 0. 15 mol/L sodium chloride, 50 mmol/L Tirs-HCl, pH 8. 0 buffer containing 3 mmol/L reduced glutathione and 1 mmol/L oxidized glutathione for 48 hours at 4 degrees C. PMID- 16706374 TI - [On the biophysics characteristics of reticulocytes]. AB - This paper reports an in vivo study on the biophysics characteristics of reticulocytes. Anemia was induced by injection of phenylhydrazine in rabbits. The measurements, including electrophoresis rate, hematolytic rate, fluorescent polarization and the changing anisotropic value, were performed in vivo for 72 hours in the process of reticulocytes growing into erythrocytes. It was shown that there were obvious changes in the biophysics characteristics of reticulocytes in this course. Therefore, the findings are of significance to basic, theoretical and clinical studies. PMID- 16706375 TI - [Construction and identification of HBD-2 transgenic mice]. AB - Human beta defensin 2 (HBD-2) may play an important role in human defense against infection. Its antimicrobial capacity has been fully documented in in vitro study. In order to evaluae its in vivo effects, we developed an HBD-2 transgenic mouse model. The HBD-2 minigene containing CMV promoter, full length of HBD-2 cDNA and BGH polyA tail was generated by PCR amplification and introduced into the fertilized oocytes of C57 X ICR hybridized mouse by microinjection, and offspring were produced. DNA was isolated from the tails of the mouse pups, and the HBD-2 minigene incorporation was analyzed by PCR using HBD-2 specific primers. The HBD-2 gene expression in the multi-tissues of transgenic mice was determined at mRNA level by RT-PCR and at peptide level by immunohistological staining with the use of HBD-2 monoclonal antibody. The results showed that among 17 F0 transgenic mice, HBD-2 positive signal was determined by PCR in 4 mice, suggesting that HBD-2 minigene has been incorporated into the offspring mice. Meanwhile, a widespread expression of HBD-2 mRNA and peptide was detected in the F1 transgenic mice's multi-tissues such as trachea, lung, intestine, esophagus, testis, spleen, skin, endothelium and brain. PMID- 16706376 TI - [Mechanical environment of rotating bioreactor and its effect on cell growth]. AB - In this paper is presented an analysis of the mechanical effect of horizontal rotating bioreactor on cell culture. Getting the microgravity of the bioreactor and the shear stress on canine mesenchymal stem cells (cMSCs) with theoretic calculating model and differential equations, we have validated the density,growth rate and modality of cultured cell by scanning electron microscopy. The horizontal rotating bioreactor which we developed could create the mechanic environment of microgravity (K<8.38 X 10(-2))and low shear stress(r<1.62 dyn/cm2) in theory. The results of scanning electron microscopy indicated that the cells' growth-speed, quantity and modality in bioreactor were better than those of cells cultured in static 24-well plate. The mechanical environment of the rotating bioreactor is propitious for keeping better modality and more rapid proliferation of cMSCs. The rotating bioreactor is a novel approach and technique it is superior to static culture. PMID- 16706377 TI - [Quantitative detection of GSTP1 gene sin the lung tissue of hypobaric hypoxia modeling rats]. AB - We have investigated the susceptibility of rat lung's GSTP1 gene to hypobaric hypoxia and explored its role in the body's possible adaptation mechanism at the moleuclar lever. Thirty male SD rats were randomly divided into five groups(0,1,3,5 and 7 d) and were exposed for 12 h per day at a simulating altitude of 7000 +/- 50 m in a hypobaric hypoxia chamber with 1 h's rest after 6 h's exposure. Then the expression of GSTP1 mRNA in the lung tissue of SD rats was examined using fluorescence quantitative RT-PCR. Meanwhile the activity of glutathione S-transferases (GSTs) enzyme and the change of maleic dialdehyde (MDA) in the lung tissue of SD rats were determined using spectrophotometer. In comparison with the non-exposure group,the expression of GSTP1 gene showed statistically significant differnce from the first to the seventh day (P<0.05). The level of GSTs decreased and MDA increased from the first to the seventh day (P<0.05). In conclusion, GSTP1 gene is susceptible to hypobaric hypoxia and may be a new marker of gene screening for the body's adaptation to special environment. PMID- 16706378 TI - [Rule induction algorithm for brain glioma using support vector machine]. AB - A new proposed data mining technique, support vector machine (SVM), is used to predict the degree of malignancy in brain glioma. Based on statistical learning theory, SVM realizes the principle of data dependent structure risk minimization, so it can depress the overfitting with better generalization performance, since the prediction in medical diagnosis often deals with a small sample. SVM based rule induction algorithm is implemented in comparison with other data mining techniques such as artificial neural networks, rule induction algorithm and fuzzy rule extraction algorithm based on fuzzy max-min neural networks (FRE-FMMNN) proposed recently. Computation results by 10 fold cross validation method show that SVM can get higher prediction accuracy than artificial neural networks and FRE-FMMNN, which implies SVM can get higher accuracy and more reliability. On the whole data sets, SVM gets one rule with the classification accuracy of 89.29%, while FRE-FMMNN gets two rules of 84. 64%, in which the rule got by SVM is of quantity relation and contains more information than the two rules by FRE-FMMNN. All the above show SVM is a potential algorithm for the medical diagnosis such as the prediction of the degree of malignancy in brain glioma. PMID- 16706379 TI - [A new MTI scheme in ultrasonic color flow mapping systems]. AB - In color flow imaging systems, the ultrasound Doppler signal scattered from blood is heavily corrupted by the strong clutter signal reflected from the slow-moving muscular tissue. Moving target indicator (MTI) is a key technique of rejecting the clutter signal and detecting blood flow. To improve the performance of detecting blood flow, a new MTI scheme is proposed in this paper. It consists of a pre-filter, a clutter weak-rejector and a 2-order AR estimator. After the processing of pre-filtering and clutter weak-rejection, the strong clutter signal is effectively eliminated and its strength is comparable with blood flow. Under such condition, the optimal performance can be attained by the 2-order AR estimator. It is illustrated in the simulation that, compared with the data from traditional methods, the blood flow parameters are estimated with smaller deviation and the lower speed flow is detected. PMID- 16706380 TI - [Computer simulation methods of cardiac electrophysiology]. AB - Computer simulation is one of the powerful protocols to study electrophysiology theoretically. In this paper,the algorithm of Rush and Larsen was used to solve the ordinary differential equations (ODE's) in the Luo-Rudy models of mammalian ventricular cell. The operator splitting and adaptive time step methods were used to solve the partial differential equations (PDE's) in cardiac tissue conduction models. Using these methods we accomplished the simulation programs of single ventricular cell model and two-dimensional (2-D) tissue model. The methods of initiating spiral waves were studied with this software. The data obtained from 2 D simulation can be used for further study on isopotential contour lines, spiral wave tip trajectories, and pseudo-ECG. PMID- 16706381 TI - [A rapid prototype fabrication method of dental splint based on 3D simulation and technology]. AB - The conventional design and fabrication of the dental splint (in orthognathic surgery) is based on the preoperative planning and model surgery so this process is of low precision and efficiency. In order to solve the problems and be up to the trend of computer-assisted surgery, we have developed a novel method to design and fabricate the dental splint--computer-generated dental splint, which is based on three-dimensional model simulation and rapid prototype technology. After the surgical planning and simulation of 3D model, we can modify the model to be superior in chewing action (functional) and overall facial appearance (aesthetic). Then, through the Boolean operation of the dental splint blank and the maxillofacial bone model the model of dental splint is formed. At last, the dental splint model is fabricated through rapid prototype machine and applied in clinic. The result indicates that, with the use of this method, the surgical precision and efficiency are improved. PMID- 16706382 TI - [Three-dimensional moston estimation of coronary artery from single-plane cineangiogram sequences]. AB - This paper presents a method for estimating three-dimensional (3D) motion of coronary arteries from single-plane X-ray angiogram sequences on two views. Firstly, original images are preprocessed and two-dimensional (2D) vessel skeletons are extracted. 2D motion estimation is performed along the skeletons in two images. Then geometrical transformation matrix between views is obtained based on perspective projection model for X-ray angiography system, and 3D coordinate of spatial points are calculated. The 3D motion estimation and reconstruction algorithm is applied along the two image sequences to accomplish 3D reconstruction of vessel skeletons and motion vectors between consecutive time instants. Its effectiveness has been demonstrated on clinical single-plane coronary artery angiograms and potential errors are discussed. PMID- 16706383 TI - [A digital subtraction angiography system based on LUT algorithms]. AB - Look-up table (LUT) algorithms have been widely used in digital signal processing, but the article on the application of LUT algorithms in digital subtraction angiography was rarely reported. In this article, the effect of different LUT algorithms on digital subtraction angiography images is introduced. The result reveals that different LUT algorithms can bring about different effects of image. Based on analysis and comparison, we deem it possible to acquire improved images of DSA by use of some LUT algorithms with image processing. PMID- 16706384 TI - [Research and development of biomedical application of carbon nanotubes and related composites]. AB - Carbon nanotubes, a new member of the carbon material family, can be considered as graphite sheets rolled-up into cylinders with diameters ranging in the nanometer scale. In recent years, carbon nanotubes have attracted intensive interests because of their unique nanostructures and outstanding mechanical, electrical and magnetic properties. In this paper, the structures and basic features of carbon nanotubes were described in brief. The research advances in the carbon nanotubes on the specific recognition of biomolecules by surface modification and functionalization, in the enhancement to cell growth as culture scaffolds in vitro, and in the improvement of biocompatibility for implantable biomedical material were reviewed. Also comments were made on their potential applications in biomedical sensor and biomedical microelectrics. PMID- 16706385 TI - [Research development and prospect of calcium phosphate biomaterials with intrinsic osteoinductivity]. AB - Calcium phosphates ceramics are frequently used as bone substitute materials in clinics because they are similar to the bone in mineral phase, they are of no antigenicity, and they have good biocompatibility as well as excellent osteoconductivity. The question as to whether calcium phosphate biomaterials possess osteoinductivity has been debated for a long time. Up to now, extensive fundamental studies and clinical application have demonstrated that calcium phosphate ceramics with special structure may induce bone formation in soft tissue. In this paper are reviewed the discovery and confirmation of calcium phosphates ceramics with intrinsic osteoinductivity, the process and mechanism of osteoinduction, and the relationship between osteoinductivity of calcium phosphate materials and animal species, the mesenchymal stem cells related to osteoinduction, the application of osteoinductive biomaterials. The researches directed toward to the osteoinduction of calcium phosphate are prospected. PMID- 16706386 TI - [Multislice CT of pulmonary arterial diseases]. AB - This paper reviews the current state of the technique of multislice CT (MSCT) scan and imaging postprocessing of CT pulmonary angiography, as well as the CT features of different pulmonary arterial diseases. Investigations demonstrate that MSCT and CTA are available means to visualize pulmonary artery structures and the anatomic correlation between pulmonary artery and adjacent structures. They are fast, effective and noninvasive methods for diagnosing pulmonary arterial diseases. PMID- 16706387 TI - [Moxibustion and its application in anti-aging study]. AB - The research on anti-aging technology is always a foreland in the field of researches into human health. Based on a review of the researches for anti-aging, this paper described all kinds of theory of aging and laid the emphasis on the status quo of researches on the use of Traditional Chinese Medicine(TCM) moxibustion for anti-aging, including the research on the theory of the moxibustion in anti-aging, the research on selecting acupoints of moxibustion for anti-aging, and the clinical application of moxibustion in anti-aging in detail. The significance of herbalist theory in anti-aging and the wider development foreground were high-lighted. PMID- 16706388 TI - [The MSCT methods and radiological features of aortic diseases]. AB - With the advancement of multi-detector spiral computer tomography (MSCT) and Z axis resolution, CT has progressively been applied in showing the anatomy and disease of vessels; therefore, CTA is increasingly important to the diagnosis of vascular diseases. In this paper we focuses on a review of the progress of MSCT and CTA in diagnosing aortic diseases. PMID- 16706389 TI - [Description and integration of biomedical information resources by metadata]. AB - With the development and utilization of computer techniques, Internet is playing an important role in information dissemination. There are abundant biomedical resources on the Internet. Accessing biomedical information is more dependent on the Internet than ever. It is important to explore new methods to describe and manage information resources. We have analyzed biomedical databases, search engines, web sites, and the metadata adopted by biomedical databases. The results show that biomedical information resources are characterized by electronic format, networking, dynamia, and dispersion. Describing a resource with metadata allows it to be understood by both humans and machines in ways that promote interoperability. Metadata interoperability has to be the underlying principle for networked information management. It directly impinges on information sharing, interchange, and accessibility across the boundaries of systems, languages, and geographic locations. We can use metadata to describe biomedical information and to integrate resources. It will benefit the people to access, select, and utilize biomedical information resources. PMID- 16706390 TI - [Current reviews in aptamers]. AB - Aptamers are oligo-nucleotides which are selected by SELEX (systematic evolution of ligands by exponential enrichment) in vitro, it can bind protein or other small molecules with high specificity and affinity. Researches on aptamers have achieved much progress in biosensor, drug development and nano technology. This article has reviewed the researches and applications of aptamers in the past two years. PMID- 16706391 TI - Infrared and electronic spectroscopy of a model system for the nucleophilic substitution intermediate in the gas phase: the C-N valence bond formation in the benzene-ammonia cluster cation. AB - Infrared and electronic spectroscopy was applied to the benzene-ammonia cluster cation in the gas phase, and the observed spectra revealed the formation of a new C-N valence bond between the benzene and ammonia moieties, which has been predicted by the quantum chemical calculations (Tachikawa, H. Phys. Chem. Chem.Phys. 2002, 4, 6018). This cluster cation is regarded as a model for the cyclohexadienyl type intermediate in nucleophilic substitution reactions. PMID- 16706392 TI - General method for the dimension reduction of adaptive control experiments. AB - Adaptive femtosecond control experiments are expanding the possibilities for using laser pulses as photophysical and photochemical reagents. However, because of the large number of variables necessary to perform these experiments (usually 100-200), it has proven difficult to elucidate the underlying control mechanisms from the optimized pulse shapes. If adaptive control is to become a widespread tool for examining chemical dynamics, methods must be developed that reveal latent control mechanisms. This manuscript presents a generally applicable method for dimension reduction of adaptive control experiments based on partial least squares regression analysis (PLS) of the normalized covariance matrix of the total data set. When applied to experimental results obtained in our laboratory, it shows that only seven fundamental dimensions from an original 208-dimension search space are needed to account for approximately 90% of the variance in the observed fitness of 11,700 laser-pulse shapes explored during the optimization experiment. Furthermore, the seven dimensions have a remarkable regularity in their functional form. It is anticipated that this work will facilitate theoretical treatments directly linking the optimal fields to control mechanisms, allow quantitative comparisons of independent control results, and suggest new experimental methods for rapid adaptive searches. PMID- 16706393 TI - Control of branching ratios in the dissociative ionization of deuterium chloride. AB - The dissociative ionization of deuterium chloride (DCl) has been investigated by employing femtosecond laser pulses at 805 nm. The product branching ratio D(+)/Cl(+) of the fragments D(+) and Cl(+) is strongly affected by the chirp alpha of the laser pulses. The ratio can be controlled by a factor of 3 ranging from D(+)/Cl(+) = 0.7 at alpha = -800 fs(2) to D(+)/Cl(+) = 1.9 at alpha = +150 fs(2). The observation can be rationalized by a model where negative chirp favors intra-electronic state excitation, and positive chirp favors inter-electronic state excitation in the dissociation of the molecular ion. Complementary experiments on hydrogen chloride (HCl) are discussed. PMID- 16706394 TI - Remarkable features in the interactions of quadrupolar molecules. AB - Quadrupolar charge fields of molecules and of molecular fragments give rise to unique features in weakly interacting clusters and aggregations. Relative to dipole-dipole interactions, the interactions among quadrupolar molecules tend to allow for greater orientational distortions away from equilibrium. Potential surface regions have been found for several clusters that are attractive and yet very flat for certain directions. There is a notable slipperiness for the interactions in some of these cases. This implies significant vibrational excursions even in the ground state. Furthermore, the coupling of rotations among nearby molecules in pure clusters of quadrupolar molecules is different than for dipolar species, and it can lead to unexpectedly small internal rotation barriers. How these and other features develop and what they might imply for materials and biomolecular simulations are discussed here. PMID- 16706395 TI - Further evidence of an inverted region in proton transfer within the benzophenone/substituted aniline contact radical ion pairs; importance of vibrational reorganization energy. AB - The dynamics of proton transfer within the triplet contact radical ion pair of a variety of substituted benzophenones with N,N-diethylaniline, N,N-dimethyl-p toluinide, and N,N-diallylaniline are examined in solvents of varying polarity. The correlation of the rate constants with driving force reveal both a normal region and an inverted region providing support for the nonadiabatic nature of proton transfer within these systems. The reorganization of both the solvent and the molecular framework are central in governing the overall reaction dynamics. PMID- 16706396 TI - Photochemical ring-opening and intramolecular hydrogen shift reactions in sulfur analogues of alpha-pyrone. AB - A combined matrix isolation FTIR and theoretical DFT(B3LYP)/6-311++G(d,p) study of the photochemistry of sulfur analogues of alpha-pyrone [2H-thiopyran-2-one (TP) and 2H-pyran-2-thione (PT)] was carried out. The vibrational spectra of monomers of the compounds isolated in low-temperature argon matrixes were studied experimentally and assigned completely on the basis of theoretical calculations. UV irradiation (lambda > 337 nm) of the studied compounds isolated in low temperature matrixes results mainly in the ring-opening reaction by means of the cleavage of the alpha-bond. Other photoprocesses, not involving the alpha-bond cleavage step (such as generation of Dewar valence isomer), correspond to the minor reaction channels in both studied compounds. The ring-opening photoreaction in PT represents the first reported case of an alpha-bond cleavage in a compound with a C=S group attached to a six-membered ring, in which the internal strain practically does not exist, whereas the corresponding reaction in TP (a cleavage of a C-S bond in the alpha position with respect to a carbonyl group) is now reported for the first time. Following the ring-opening reactions, isomerization processes and intramolecular hydrogen shift reactions were observed, enabling production of TP from PT and vice versa. A detailed study of such processes was undertaken, and kinetical and mechanistical data are presented and discussed. PMID- 16706397 TI - CASSCF/CAS-PT2 study of hole transfer in stacked DNA nucleobases. AB - CASSCF and CAS-PT2 calculations are performed for the ground and excited states of radical cations consisting of two and three nucleobases. The generalized Mulliken-Hush approach is employed for estimating electronic couplings for hole transfer in the pi-stacks. We compare the CASSCF results with data obtained within Koopmans' approximation. The calculations show that an excess charge in the ground and excited states in the systems is quite localized on a single base both at the CASSCF level and in Koopmans' picture. However, the CASSCF calculations point to a larger degree of localization and, in line with this, smaller transition dipole moments. The agreement between the CAS-PT2 corrected energy gaps and the values estimated with Koopmans' theorem is better, with the CAS-PT2 calculations giving somewhat smaller gaps. Overall, both factors result in smaller CASSCF/CAS-PT2 couplings, which are reduced by up to 40% of the couplings calculated using Koopmans' approximation. The tabulated data can be used as benchmark values for the electronic couplings of stacked nucleobases. For the base trimers, comparison of the results obtained within two- and three-state models show that the multistate treatment should be applied to derive reliable estimates. Finally, the superexchange approach to estimate the donor acceptor electronic coupling in the stacks GAG and GTG is considered. PMID- 16706398 TI - Investigation of the two-photon absorption cross-section in perylene tetracarboxylic derivatives: nonlinear spectra and molecular structure. AB - We investigated the 2PA absorption spectrum of a family of perylene tetracarboxylic derivatives (PTCDs): bis(benzimidazo)perylene (AzoPTCD), bis(benzimidazo)thioperylene (Monothio BZP), n-pentylimidobenzimidazoperylene (PazoPTCD), and bis(n-butylimido)perylene (BuPTCD). These compounds present extremely high two-photon absorption, which makes them attractive for applications in photonics devices. The two-photon absorption cross-section spectra of perylene derivatives obtained via Z-scan technique were fitted by means of a sum-over-states (SOS) model, which described with accuracy the different regions of the 2PA cross-section spectra. Frontier molecular orbital calculations show that all molecules present similar features, indicating that nonlinear optical properties in PTCDs are mainly determined by the central portion of the molecule, with minimal effect from the lateral side groups. In general, our results pointed out that the differences in the 2PA cross-sections among the compounds are mainly due to the nonlinearity resonance enhancement. PMID- 16706399 TI - Experimental and theoretical investigation of vibrational overtones of glycolic acid and its hydrogen bonding interactions with water. AB - The present work reports observations of the 4nu(1) and 4nu(2) O-H stretching transitions in glycolic acid, CH(2)OHCOOH, using a highly sensitive cavity ring down spectrometer. Experimental and theoretical values for the harmonic frequencies and anharmonic constants of both O-H stretching transitions were extracted and are compared with theoretical calculations in the literature. Calculations of anharmonic frequencies, intensities, and relative energies have been performed and are presented for three conformers of glycolic acid. In the presence of water, an interesting broad spectral feature appeared underneath 4nu(1) and 4nu(2). New calculations for harmonic frequencies, intensities, and relative energies of four CH(2)OHCOOH-H(2)O complexes are reported to aid in understanding the observed spectrum. This work suggests that the perturbations are caused by intermolecular hydrogen bonding of glycolic acid with one or more water molecules. PMID- 16706400 TI - Reactivity of the CHBr2+ dication toward molecular hydrogen. AB - Structural aspects as well as the stability and reactivity of the CHBr(2+) dication are studied both experimentally and theoretically. Translational energy distributions of the CHBr(+) products from charge transfer between CHBr(2+) and Kr indicate that the dication exists in two isomeric forms, H-C-Br(2+) and C-Br H(2+). In the reaction of CHBr(2+) with H(2), the dominant channel corresponds to proton transfer leading to CBr(+) + H(3)(+). Other reaction channels involve the formation of the intermediates CH(3)Br(2+) and CH(2)BrH(2+), respectively. Both of the latter dications can either lose a proton to form CH(2)Br(+) or undergo a spin-isomerization followed by cleavage of the C-Br bond. The proposed mechanisms are supported by DFT calculations and deuterium labeling experiments. PMID- 16706401 TI - Single site electronic spectroscopy of magnesium isobacteriochlorin in n-octane matrixes at 7 K. AB - The high resolution, single site emission and absorption spectra of magnesium isobacteriochlorin (MgiBC) in n-octane matrixes at 7 K are reported. Its emission and Q and Soret band absorption regions have been investigated. The vibrational frequencies of the ground and the lowest energy pipi* singlet excited states were determined from luminescence and excitation spectra, respectively. The emission from MgiBC at 7 K seems to be entirely fluorescence. The luminescence and Q(y) region spectra of the complex are similar, having intense, narrow origin bands followed by relatively weak, but orderly vibrational structure. The Q(x) region does not have a clear origin and exhibits complex vibrational structure that increases in intensity going to higher energy. In the Soret region the individual pipi* origins are clearly identifiable and some vibrational structure was also observed. PMID- 16706402 TI - One- and two-photon absorption and emission properties of a Zn(II) chemosensor. AB - This paper presents the synthesis and two photon-induced absorption (TPA) properties of a functionalized distyrylbenzene (DSB) 1 containing a tetra azacyclododecane (cyclen) receptor for Zn(II). The influence of Zn(II) on one- and two-photon absorption characteristics of 1 has been investigated in dimethyl sulfoxide. The experiments show that the TPA action spectrum of uncomplexed 1, at 750 nm employing nanosecond-long excitation pulses, is 5 times more intense than that of the complexed form. This moderate contrast between the bound and unbound species confirms, however, the potential of this design scheme for the development of molecular structures with enhanced sensitivity and contrast to be used as Zn(II) sensors through TPA-induced fluorescence microscopy. PMID- 16706403 TI - Effect of aggregation on the excited-state electronic structure of perylene studied by transient absorption spectroscopy. AB - The effect of aggregation on the excited-state electronic structure of perylene was studied through transient absorption measurements of isolated molecules, excimers, monomeric crystals (beta-perylene), and dimeric crystals (alpha perylene). Changes of electronic state were clearly identified from the changes in transient absorption spectra. A detailed investigation was made by combining the obtained results with previous measurements of ground-state absorption and fluorescence spectra. The energy level of the ion-pair state in alpha-perylene crystals was estimated, and the results are compared with previous photoconductivity results. Moreover, the relaxation processes of excited states in alpha-perylene crystals were studied by femtosecond transient absorption measurements. PMID- 16706404 TI - A combined matrix isolation and ab initio study of bromine oxides. AB - Bromine oxides have been generated by passing a mixture of Br(2)/O(2)/Ar through a microwave discharge. The products were stabilized at 6.5 K in an excess amount of argon. Infrared spectroscopy was used to analyze the species formed; experiments with enriched (18)O(2) and ab initio calculations were carried out to assist in the assignment of the spectra. Besides the known species BrO, OBrO, and BrBrO, spectroscopic evidence for BrOBrO, BrBrO(2), and a new isomer of Br(2)O(3) is reported for the first time. Extensive comparisons are drawn between the present studies and previous experimental and theoretical works. The chemistry involved in the production of the observed compounds is discussed. The assignments are corroborated by the good correlation between observed and calculated band positions and intensities. PMID- 16706405 TI - Electron affinities of polycyclic aromatic hydrocarbons by means of B3LYP/6-31+G* calculations. AB - The gas-phase experimental adiabatic electron affinities (AEAs) of the polycyclic aromatic hydrocarbons (PAHs) anthracene, tetracene, pentacene, chrysene, pyrene, benzo[a]pyrene, benzo[e]pyrene, and fluoranthene are well reproduced using the hybrid density functional method B3LYP with the 6-31+G* basis set, indicating that the smallest addition of diffuse functions to the basis set is suitable for a correct description of the stable PAH anion states. The calculated AEAs also give a very good linear correlation with available reduction potentials measured in solution. The AEAs (not experimentally available) of the isomeric benzo[ghi]fluoranthene and cyclopenta[cd]pyrene, commonly found in the environment, are predicted to be 0.817 and 1.108 eV, respectively, confirming the enhancement of the electron-acceptor properties associated with fusion of a peripheral cyclopenta ring. The calculated localization properties of the lowest unoccupied MO of cyclopenta[cd]pyrene, together with its relatively high electron affinity, account for a high reactivity at the ethene double bond of this PAH in reductive processes. PMID- 16706406 TI - Classical polarization in hybrid QM/MM methods. AB - We have presented a method for modeling polarization in hybrid QM/MM calculations. The method, which expresses the induced dipoles as a set of "induced" charges, is based on the induced dipole approach and methodology for calculating potential-derived point charges from distributed multipole series. The method has the advantage that the same methodology can be used to determine the induced charges and the potential derived charges and so both sets of charges are rigorously defined within the same framework. This underlying link with the wave function makes the method particularly suitable for use in hybrid QM/MM calculations. Here we assess the importance of explicit polarization in the classical part of a QM/MM system with regard to improving the classical description and the consequent effects on the quantum description. The main advantages of the induced charge approach are that the method is readily interfaced with quantum mechanical methods and that induced charges are more readily interpreted than induced dipoles. The ease of interpretation is illustrated by analysis of the charges involved in dimeric and trimeric hydrogen bonded systems. The method for treating the MM polarization has been validated by a regression analysis of the charges induced in both the QM and MM systems against those derived from full quantum mechanical calculations. The method has also been validated using two energy decomposition approaches, which show that MM polarization makes a significant and reliable contribution to the QM - MM interaction energy in a hybrid system. The distance dependency of the induced charges is investigated in calculations on methylsuccinyl-Ala-Ala-Pro-Ala chlormethyl ketone interacting with human neutrophil elastase and propranolol interacting with asparagine residues in a model of the beta(2)-adrenergic receptor. PMID- 16706407 TI - Computation of relative bond dissociation enthalpies (DeltaBDE) of phenolic antioxidants from quantum topological molecular similarity (QTMS). AB - A recently proposed method called quantitative topological molecular similarity (QTMS) generated a model for the computation of the relative substituent effects on the bond dissociation enthalpies (DeltaBDEs) for a set of 39 phenols. The data set includes a diverse set of substituents with monosubstituted and poly substituted derivatives that exhibit different electronic and steric effects. Many share common structural features with already well-established antioxidants. QTMS reveals the active region of the substituted phenols and identifies the electronic descriptors that best explain the range of DeltaBDEs observed. For substituents in the 4-X position (para) we find that our model requires a correction for radical stabilization enthalpy (RSE). Application of the QTMS methodology yields an unrivalled QSAR with r(2) = 0.98 and q(2) = 0.85 for the bond dissociation enthalpies of this phenolic antioxidant data set. PMID- 16706408 TI - Study of rhamnose radicals in the solid state adopting a density functional theory cluster approach. AB - A theoretical study is performed on the radiation-induced radicals in crystalline alpha-l-rhamnose, using density functional theory (DFT) calculations. Irrespective of earlier structural assignments, a host of possible radical models is examined in search for a structure that accurately reproduces experimental electron paramagnetic resonance (EPR) properties. A cluster approach is followed, incorporating all hydrogen bond interactions between radical and crystalline environment. Hyperfine coupling tensors as well as g tensors are determined and a comparison is made with available experimental data. Three carbon-centered hydroxyalkyl radicals are validated, in accordance with experimental suggestions for their structure. The occurrence of a carbon-centered oxygen anion radical for one of the radical species is rejected on theoretical grounds, and instead an altered hydroxyalkyl structure is suggested. Our cluster calculations are able to determine g and hyperfine tensors for the oxygen-centered alkoxy radical in rhamnose, in accordance with one of the two measurements for this species. For all radical models, quantitative agreement with experimental hyperfine tensors is obtained by performing full cluster DFT calculations. The inclusion of the molecular environment for the determination of this EPR property proved to be essential. PMID- 16706409 TI - Cooperativity in hydrogen-bonded interactions: ab initio and "atoms in molecules" analyses. AB - The H(2)CO...(HF)(n) (n = 1, ..., 9) complexes were investigated using the MP2 method and the following basis sets: 6-311++G(d,p), aug-cc-pVDZ and aug-cc-pVTZ. It was found that the cooperativity effect enhances significantly the F-H...O hydrogen bond; in some of cases one can detect the covalent nature of hydrogen bonding. To deepen the nature of the interactions investigated, the scheme of decomposition of the interaction energy was applied; for stronger H-bonds where the cooperativity is more important, the delocalization energy term increases. The ratio of delocalization energy to electrostatic energy increases for stronger hydrogen bonds where the proton...acceptor distance is shorter. The Bader theory was also applied, and it was found that for stronger H-bonds the electronic energy density at the proton...acceptor bond critical point is negative and may be attributed to the partly covalent interaction. PMID- 16706410 TI - Theoretical and experimental investigation of the energetics of cis-trans proline isomerization in peptide models. AB - The energetics of cis-trans proline isomerization in small peptide models have been investigated using the hybrid density functional theory method B3LYP with a 6-31+G* basis set. The molecules studied are models for the phospho-Ser/Thr-Pro substrate for Pin-1, a peptidyl-prolyl isomerase (PPIase) involved in cell division. Pin-1 requires phosphorylation of a Ser or Thr residue adjacent to a Pro residue in the substrate and catalyzes cis-trans isomerization about the proline amide bond. The dihedral angle that would correspond to the reaction coordinate for isomerization of the omega peptide bond was investigated for several small models. Relaxed potential energy scans for this dihedral angle in N methylacetamide, 1, N,N-dimethylacetamide, 2, acetylpyrrolidine, 3 and acetylproline, 4, were carried out in 20 degrees steps using the B3LYP/6-31+G* level of theory. In addition, similar scans were carried out for 1-4 protonated on the acetylamide carbonyl oxygen. Optimized structures for 1-4 protonated on the amide nitrogen were also obtained at B3LYP/6-31+G*. Relative proton affinities were determined for each site at various angles along the reaction coordinate for isomerization. The relative proton affinities were anchored to experimental gas phase proton affinities, which were taken from the literature for 1 and 2, or determined in an electrospray ionization-quadrupole ion trap instrument using the extended kinetic method for 3 and 4. Proton affinities of 925 +/- 10 and 911 +/- 12 kJ/mol were determined for 3 and 4, respectively. These studies suggest that the nitrogen atom in these amides becomes the most basic site in the molecule at a dihedral angle of ca. 130 degrees . In addition, the nitrogen atoms in 2-4 are predicted to attain basicities in the range 920-950 kJ/mol, making them basic enough to be the preferred site for hydrogen bonding in the Pin-1 active site, in support of the proposed mechanism for PPIases. PMID- 16706411 TI - Conformational behavior of dimethyl 5-methyl-1H,3H-pyrrolo[1,2-c][1,3]thiazole 6,7-dicarboxylate 2,2-dioxide isolated in low-temperature matrixes. AB - The structure of dimethyl 5-methyl-1H,3H-pyrrolo[1,2-c][1,3]thiazole-6,7 dicarboxylate 2,2-dioxide (PTD) was investigated in low-temperature noble gas matrixes (Ar, Kr, Xe), amorphous solid, and the crystalline state by infrared spectroscopy and computational methods. The geometry of PTD conformers is defined by the orientation of two methyl ester groups, which may adopt pseudo-trans or pseudo-cis positions in relation to the pyrrolo-thiazole system. For both methyl ester groups, the latter arrangement was predicted by the calculations to be energetically the most favorable in the isolated molecule. The envelope form of the thiazolidine ring is present in all conformers, with the sulfur atom placed in the apex position, while the pyrrole ring is almost planar. Three types of conformers differing in the orientation of the methyl ester groups relative to the pyrrolo-thiazole system (cis/cis, trans/cis, cis/trans) were identified in the matrixes. The cis/cis forms were found to be the most stable ones in both gaseous state and argon matrixes. On the other hand, the more polar trans/cis forms were found to be stabilized in the more polarizable krypton and xenon matrixes as well as in the neat amorphous and crystalline phases. On the basis of annealing experiments, performed in argon and xenon matrixes up to 35 and 68 K, respectively, conformational changes preceding the aggregation of the compound are suggested. PMID- 16706412 TI - pKa prediction using group philicity. AB - Acid-base dissociation constants (pK(a) values) are important in understanding the chemical, environmental and toxicological properties of molecules. Though various methods have been developed to predict pK(a) by experimental and theoretical models, prediction of pK(a) is still complicated. Hence, a new approach for predicting pK(a) using the group philicity concept has been attempted. Presence of known functional groups in a molecule is utilized as the most important indicator to predict pK(a). The power of this descriptor in describing pK(a) for the series of carboxylic acids, various substituted phenols, anilines, phosphoric acids, and alcohols is probed. Results reveal that the group electrophilicity is suitable for effectively predicting the pK(a) values. PMID- 16706413 TI - Estimation of the barrier to rotation of benzene in the (eta 6-C6H6)2Cr crystal via topological analysis of the electron density distribution function. AB - The high-resolution X-ray diffraction analysis of the electron density distribution and plane-wave density functional theory has been applied to estimate the lattice energy and barrier to rotation of a benzene ring in the crystal of (eta(6)-C(6)H(6))(2)Cr. Experimental data made it possible to perform analysis of the metal-(pi-ligand) bond and estimate the nature and energy of weak H...H and H...C intermolecular interactions in the crystal. Summation of the intermolecular H...H and H...C interaction energies makes it possible to reproduce the experimental sublimation enthalpy value with high accuracy. PMID- 16706414 TI - EPR and ENDOR study of radiation-induced radical formation in purines: sodium inosine crystals X-irradiated at 10 K. AB - X-irradiated single crystals of sodium inosine (Na(+)*Inosine(-)*2.5H(2)O), in which the hypoxanthine base is present as the N1-deprotonated anion, were investigated using K-band (24 GHz) electron paramagnetic resonance (EPR), electron-nuclear double resonance (ENDOR), and ENDOR induced EPR (EIE) techniques at 10 K. At least five different radicals were present immediately after irradiation at 10 K. R1, which decayed upon warming the crystals to 50 K, was identified as the electron-loss product of the parent N1-deprotonated hypoxanthine base. Hyperfine couplings to HC8 and HC2 were fully characterized with ENDOR spectroscopy, and the identification was supported by DFT calculations. R2, which also decayed on warming to 50 K, exhibited nearly equal couplings to HC2 and HC8. Taken in combination with an extensive set of DFT calculations, the experimental results indicate that R2 is the (doubly negative) product of electron-gain by the initially anionic N1-deprotonated hypoxanthine parent. R3, which exhibited hyperfine coupling only to HC8 could not be identified. R4, which persisted on annealing to 260 K, exhibited one large alpha proton hyperfine coupling which was fully characterized by ENDOR. Based on DFT calculations and the experimental data, R4 was identified as the product of net H abstraction from C5'. The remaining HC5' was the source of the measured alpha proton coupling. R5, present at low temperature and the only observable radical after warming the crystals to room temperature, was identified as the C8-H addition radical. The alpha-coupling to HC2 and beta-couplings to the pair of C8 methlyene protons were fully characterized by ENDOR. PMID- 16706415 TI - Theoretical study of alkali cation-benzene complexes: potential energy surfaces and binding energies with improved results for rubidium and cesium. AB - High level ab initio quantum chemical calculations have been carried out on the binding of alkali metal to benzene with special attention to heavier metals for which the agreement between the most recent theoretical investigations and the experimental bond dissociation energies (BDEs) is not very good. We performed BSSE-corrected geometry optimizations employing the MP2 level of theory with large basis sets and a modified Stuttgart RSC 1997 basis set for rubidium and cesium and carried out single point energy calculations at the MP4 level, obtaining, also for the latter metals, BDE values in good agreement with the experimental results. Furthermore, in view of the development of empirical correction terms to force fields to describe cation-pi interactions, we evaluated the potential energy surface along the benzene symmetry axis and discussed the role of the BSSE correction on the accuracy of our results. PMID- 16706416 TI - Space-time contours to treat the interaction between an intense electric field and a molecular system. AB - A few years ago, we developed an approach to treat molecular systems exposed to an external, intense, time-dependent field (J. Phys. Chem. A 2003, 107, 4724; J. Chem. Phys. 2003, 119, 6998). Within this study, we encountered two novel concepts: the dressed (namely, field affected) time-dependent nonadiabatic coupling term and the space-time contours. In the present article, we analyze the newly introduced nonadiabatic coupling term and discuss its importance for dynamical studies. We also refer to the just mentioned space-time contour and present the more efficient contour for realistic situations. The scope of the above-mentioned articles is extended with the aim of defining quasi-adiabatic states for such situations. Strictly speaking, molecular systems exposed to intense, fast oscillating fields are not expected to form adiabatic states. Still we consider such a situation and end up with three possibilities for quasi adiabatical time-dependent states eventually to be used within the Born Oppenheimer approximation. PMID- 16706417 TI - Effect of sample preparation and excitation conditions on the single molecule spectroscopy of conjugated polymers. AB - Extensive new single molecule spectroscopy (SMS) data on the conjugated polymer MEH-PPV at low temperature were obtained. In particular, the combined effects of sample preparation and excitation condition were explored in detail. The data confirm previous observations from this laboratory that (i) the distribution of emission maxima of single MEH-PPV molecules has a bimodal distribution and (ii) the single molecule emission spectrum of MEH-PPV exhibits few time-dependent fluctuations of the emission intensity, band shape, or spectral maxima. These data also help explain the discrepancy among the various published SMS data on this compound and suggest that environmental impurities, long irradiation times, nearby interfaces, and incomplete data sampling may account for some of the discrepancies among the published data. PMID- 16706418 TI - Observation of triatomic species with conflicting aromaticity: AlSi2- and AlGe2-. AB - We created mixed triatomic clusters, AlCGe(-), AlSi(2)(-), and AlGe(2)(-), and studied their electronic structure and chemical bonding using photoelectron spectroscopy and ab initio calculations. Excellent agreement between theoretical and experimental photoelectron spectra confirmed the predicted global minimum structures for these species. Chemical bonding analysis revealed that the AlSi(2)(-) and AlGe(2)(-) anions can be described as species with conflicting (sigma-antiaromatic and pi-aromatic) aromaticity. The AlCGe(-) anion represents an interesting example of chemical species which is between classical and aromatic. PMID- 16706419 TI - Orientational correlations in the glacial state of triphenyl phosphite. AB - Spallation neutron and high-energy X-ray diffraction experiments have been performed to investigate the local structure of the glacial and supercooled liquid states in triphenyl phosphite. The observed diffraction patterns have been interpreted using a Reverse Monte Carlo modeling technique. The results show that the glacial state forms unusually weak intermolecular hydrogen bonds between an oxygen atom connected to a phenyl ring and an adjacent phenyl ring aligned in an approximately antiparallel configuration. The structure is very different from the hexagonal crystal which is characterized by two weaker hydrogen bonds between linear arrays of molecules which are offset from each other and packed in a hexamer arrangement. PMID- 16706420 TI - Versatile fabrication of distorted cubic mesoporous silica film using CTAB together with a hydrophobic organic additive. AB - We have succeeded in the fabrication of distorted cubic R-3m mesoporous silica film with 3D open mesostructure providing high pore accessibility from the surface-air interface of the film. The film fabrication involves a unique approach of adding 1,3,5- triisopropylbenzene (TIPB) to the silica/cetyltrimethylammoniumbromide (CTAB) coating sol during its preparation. The addition of TIPB induces the formation of spheroid micelles, promoting R-3m mesostructure in the film. This phase does not exist in the corresponding binary phase diagram of CTAB surfactant. PMID- 16706421 TI - Pressure tuning between NH...N hydrogen-bonded ice analogue and NH...Br polar dabcoHBr complexes. AB - At normal conditions 1,4-diazabicyclo[2.2.2]octane hydrobromide [C(6)H(13)N(2)](+.)Br(-) forms centrosymmetric crystals, space group Pm2, NH(+)...N hydrogen-bonded linear polycationic chains with disordered protons in the structure. As in H(2)O ice Ih, the protons in [C(6)H(13)N(2)](+.)Br(-) crystals remain disordered at low temperatures. Above 0.4 GPa the [C(6)H(13)N(2)](+.)Br(-) crystals transform into a new polar NH(+)...Br(-) hydrogen bonded complex, space group Cmc2. It has been crystallized in-situ in a diamond anvil cell and its structure determined by X-rays. The low-pressure triggering of this transformation indicates that it is a possible source of defects in the real structure at normal conditions, where, along with disproportionation defects, they can be responsible for anomalous dielectric properties, including relaxor-like behavior of NH...N hydrogen-bonded compounds. PMID- 16706422 TI - Periodic inclusion of room-temperature-ferromagnetic metal phosphide nanoparticles in carbon nanotubes. AB - We demonstrate the use of sequential catalytic growth to encapsulate iron, nickel iron, and iron-cobalt phosphide catalyst nanoparticles periodically along the entire lengths of carbon nanotubes. Investigations by local electron spectroscopies and electron diffraction reveal the compositions and crystal structures of the encapsulated particles. Significantly, high spatial resolution magnetic characterization using magnetic force microscopy and off-axis electron holography demonstrates that encapsulated iron-cobalt phosphide nanoparticles are ferromagnetic at room temperature, in accordance with the properties of bulk metal phosphides of the same structure and composition. PMID- 16706423 TI - Quantum sieving effect of modified activated carbon fibers on H2 and D2 adsorption at 20 K. AB - Quantum sieving of activated carbon fibers (ACFs) and their fluorides was observed for H(2) and D(2) adsorption at 20 K. Fluorination reduced the slit shaped pore width of ACFs by 0.2 nm. The activated carbon fibers can act as highly efficient quantum sieves for H(2) and D(2), because the effective size of an H(2) molecule is larger than that of a D(2) molecule due to the uncertainty principle and the molecular size difference between H(2) and D(2) is significant in the micropore space. The D(2)/H(2) selectivity of ACFs evaluated by ideal adsorption solution theory was larger than that of the fluorinated ACFs. PMID- 16706424 TI - 57Fe Mossbauer spectroscopy on multiwalled carbon nanotubes with metal filling. AB - Mossbauer measurements at different temperatures are reported for MWCNTs with metallic encapsulations ("nanowires"). The spectra can reasonably be refined with two subpatterns: one providing clear evidence of an iron carbide Fe(3)C (cementite) phase as the main nanowire component and the other yielding a relaxation doublet most probably belonging to the same phase. Whereas the former one displays a well resolved magnetic hyperfine spectrum with Brillouin type temperature dependence, the latter one gains importance with rising temperature or onset of an inhomogeneous external magnetic field. The comparably large incoherent scattering is attributed to the graphene layers of the tube walls. The experimental results are discussed on the background of an interpretation model trying to explain unusual magnetometric results published elsewhere. PMID- 16706425 TI - Nanoconfinement and the glass transition: a cluster hypothesis. AB - It is suggested that the depression of the glass temperature in nanoconfined liquids and polymers may be explained by an effect of the confinement alone: specifically, confinement by hard walls creates an excluded volume effect that decreases the fraction of molecules organized in clusters. If we think of this fraction as a rough surrogate determining the fictive or structural temperature, then the confined liquid will have a higher fictive temperature than the bulk liquid at the same actual temperature. A computational method already used to study clusters may test the hypothesis. PMID- 16706426 TI - Surfactant transfer through a liquid membrane: origin of spontaneous oscillations at the membrane/acceptor phase interface. AB - Instability due to surfactant redistribution in a liquid membrane system consisting of two solutions, namely source and acceptor, separated by a layer of immiscible liquid is studied theoretically and experimentally. The transfer of a surfactant from a source phase to an acceptor phase is often accompanied by spontaneous nonlinear oscillations of electrical potential and/or interfacial tension. The oscillations can be generated at each of the membrane interfaces. Here a mechanism of oscillation, which develops at the membrane/acceptor phase interface, is proposed on the basis of direct numerical simulation of the system evolution. Performed experimental studies confirm the theoretical results. PMID- 16706427 TI - Limitations of the methyl radical recombination method for acoustic cavitation bubble temperature measurements in aqueous solutions. AB - The methyl radical recombination (MRR) method has been used for the measurement of cavitation bubble temperatures in aqueous solutions containing a select group of aromatic hydrocarbons as the source for the methyl radicals. The aromatic solutes used were phenol, aniline, m-cresol, and o-toluidine. The maximum bubble core temperatures determined using aniline and phenol were observed to be comparatively high with respect to other reported literature methods and also where the methyl radicals were produced from the cavitation thermolysis of simple aliphatic alcohols. It is concluded that the MRR method cannot be used with organic compounds that do not predominantly produce methyl radicals on the thermal decomposition of the hydrocarbon solutes within the hot core of a collapsing bubble. PMID- 16706428 TI - Photovoltaic properties of Au/beta-carotene/n-Si organic solar cells. AB - Photovoltaic properties of Au/beta-carotene/n-Si organic solar cells characterized by current-voltage and capacitance-voltage measurements have been investigated. The photocurrent in the reverse direction increases with increasing illumination intensity. The I(sc) increases linearly with light intensity. The I(sc) dependence of light intensity follows a power law I(sc) approximately F(alpha). The exponent alpha was found to be 1.38. This indicates a monomolecular recombination in this device. Au/beta-carotene/n-Si organic solar cells give an open-circuit voltage of 0.316 V and a short-circuit current of 2.33 x 10(-4) A at light intensity of 6 W/m(2). The best conversion efficiency for Au/beta carotene/n-Si solar cells was found to be 23.3% at a light intensity of 6 W/m(2). PMID- 16706429 TI - A reverse cation-exchange route to hollow PbSe nanospheres evolving from Se/Ag2Se core/shell colloids. AB - A reverse cation-exchange approach for the synthesis of hollow PbSe nanospheres is successfully established. This route involves a new strategy of a stepwise, in situ template-based evolution from spherical amorphous Se colloids to Se/Ag(2)Se core/shell colloids, then to hollow PbSe nanospheres. Se colloids are prepared as the initial product by utilizing the chelation of ethylenediamine to bulk Se. They are converted into Se/Ag(2)Se core/shell colloids through the reaction with Ag(+) in ethylene glycol. During the conversion from Ag(2)Se shell to PbSe shell, a small amount of tributylphosphine is crucial as the capping agent. The characterization results, including XRD, SEM, TEM, HRTEM, and EDX, reveal that hollow PbSe nanospheres with polycrystalline and cubic structure are prepared. The corresponding optical band gap is calculated to be 0.56 eV. This conformation is potentially beneficial to the improvement concerning the applications of PbSe nanostructures. PMID- 16706430 TI - Location of spectroscopic probes in self-aggregating assemblies. I. The case for 5-doxylstearic acid methyl ester serving as a benchmark spectroscopic probe to study micelles. AB - A strategy to locate spectroscopic probes in micelles is presented which involves establishing a "benchmark" probe, i.e., one whose position is well-known and against which other probe positions may be established. Theoretically calculated values of the fraction of the micelle polar shell occupied by water, H(shell), are compared with experimental values measured with the spin probe 5-doxylstearic acid methyl ester (5DSE) for a series of sodium n-alkyl sulfate micelles as functions of both the aggregation numbers and the alkyl chain length. The theoretical values involve one adjustable parameter that may be taken to be the volume in the polar shell inaccessible to water, V(dry). Under the hypothesis that the thickness of the polar shell (5 Angstroms) remains constant as either the aggregation number or the chain length is varied, we find excellent agreement between the theoretical predictions and the experimental results, using the same value of V(dry) for chain lengths 8-12 and for aggregation numbers varying from approximately 38 to 130. We argue that these are compelling reasons that 5DSE follows the zero-order model (ZOM) of probe location. The ZOM applies to any probe that rapidly diffuses within the confines of the micelle polar shell and nowhere else. Thus, 5DSE can serve as a benchmark in the sodium alkyl sulfate micelles. As a further check, results are also presented for ammonium dodecyl sulfate micelles, where 5DSE is also found to follow the ZOM, i.e, no further adjustable parameters are needed to pass from the sodium alkyl sulfate micelles to ammonium dodecyl sulfate micelles. In contrast, results are also presented for a similar spin probe 16-doxylstearic acid methyl ester (16DSE) that is found not to adhere to the ZOM in any of the micelles. A simple first-order correction to the ZOM in which 16DSE is displaced slightly from the polar shell is shown to account for the results well. The necessary displacements, which range from about 0.7 Angstroms outside the polar shell to 1.3 Angstroms inside, are not correlated with either chain lengths or aggregation numbers; however, they correlate rather well with H(shell). Calibrations of 6-, 7-, 10-, and 12DSE spin probes are presented in the Appendix, making them available to measure microviscosities and effective water concentrations. PMID- 16706431 TI - A reinterpretation of the hydration of micelles of dodecyltrimethylammonium bromide and chloride in aqueous solution. AB - The hydration of dodecyltrimethylammonium (DTAB) micelles is reinterpreted in light of the results of the companion paper (immediately preceding this paper) that showed that the location of the spin probe 16-doxylstearic acid methyl ester (16DSE) changes as a function of the aggregation number, N, of anionic micelles, i.e, that it does not conform to the zero-order model (ZOM). The ZOM requires that the NO(*) moiety diffuse throughout the Stern layer of the micelle and nowhere else. By using the ZOM as a working hypothesis, the previous interpretation (J. Phys. Chem. B 2002, 106, 1926) of 16DSE data proposed that an increasing number of alkyl chain methyl groups occupied the Stern layer as N increased. In this work, the spin probe 5-doxylstearic acid methyl ester that was found to fulfill the ZOM in anionic micelles was measured as a function of N in DTAB and was found to obey the ZOM in this cationic micelle as well. Thus, a simple model of the hydration of micelles that is successful in anionic micelles is also successful in DTAB. The previous results for 16DSE are reinterpreted here as being due to small displacements of the NO(*) moiety as a function of N. PMID- 16706432 TI - Strong two-photon absorption in new asymmetrically substituted porphyrins: interference between charge-transfer and intermediate-resonance pathways. AB - We study two-photon absorption (2PA) in two series of new free-base porphyrins with 4-(diphenylamino)stilbene or 4,4'-bis-(diphenylamino)stilbene (BDPAS) attached via pi-conjugating linkers at the porphyrin meso-position. We show that this new substitution modality increases the 2PA cross section in the Soret band region (excitation wavelength 750-900 nm) of the core porphyrin by nearly 2 orders of magnitude, from sigma(2) approximately 10 GM for the meso-phenyl substituted analogue to sigma(2) approximately 10(3) GM for the ethynyl-linked BDPAS-porphyrin dyad. The 2PA properties are quantitatively described by considering two different and interfering 2PA quantum transition pathways. The first path involves virtual transition via intermediate one-photon resonance. The second path bypasses the intermediate resonance and occurs due to a large permanent dipole moment difference between the ground and the final electronic states. To our best knowledge, this is the first experimental observation of the combined effect of these two pathways on one particular two-photon transition, resulting in quantum-interference-modulated 2PA strength. PMID- 16706433 TI - Critical evaluation of micellization behavior of nonionic surfactant MEGA 10 in comparison with ionic surfactant tetradecyltriphenylphosphonium bromide studied by microcalorimetric method in aqueous medium. AB - The micellization behavior of MEGA 10 has been studied at nine different temperatures by isothermal titration calorimetry (ITC), and thermodynamics of the process have been evaluated and examined in detail. The aggregation number of the nonionic surfactant has been estimated from the ITC results by a simulation procedure based on the mass action principle of micellization of the surfactant. The cmc of MEGA 10 has shown a minimum in temperature dependence as observed for ionic surfactants. For a comparison, the cmc and related thermodynamic parameters of an ionic surfactant, tetradecyltriphenylphosphonium bromide (C(14)TPB) studied at several temperatures in aqueous medium has been considered. The contributions of the headgroups of both the surfactants to the free energies of their respective micellization have been deciphered and presented. PMID- 16706434 TI - Enhanced stabilization and deposition of Pt nanocrystals on carbon by dumbbell like polyethyleniminated poly(oxypropylene)diamine. AB - Pseudo-dendritic polyethyleniminated poly(oxypropylene)diamine (D400(EI)(20)) was used as a stabilizer and promoter to prepare Pt nanoparticles in aqueous solution, which was then deposited on carbon surface followed by calcination. After being deposited on carbon surface, no Pt(0) could be detected in the solution phase. In all steps, the increasing molar ratio of the amino groups of D400(EI)(20) to H(2)PtCl(6) ([N]/[Pt]) drastically reduced the size and the polydispersity and kept a constant low value after [N]/[Pt] = 20. Under a [N]/[Pt] ratio of 20, the particle sizes obtained from transmission electron microscopy (TEM) were very small in solution (2.7-2.4 nm) and remained the same after being deposited on carbon surface (2.7-2.4 nm), and were only slightly increased to 3.6-3.0 nm after calcination. The stabilizing ability of D400(EI)(20) to Pt on carbon surface before and after calcination can be interpreted by the existence of binding energy between Pt and amine nitrogen. The X-ray diffraction (XRD) pattern together with the TEM image reveals that the obtained Pt nanoparticles exist in single-crystal form. The results of photoelectron spectroscopy (XPS) evidence that the metallic Pt(0) rather than the oxidized Pt is the predominant species in the Pt/C catalysts. The electrochemical active surface (EAS) area of the Pt/C catalyst is only slightly higher than that of the E-TEK Pt/C catalyst, but the utilization factor (93.4%) is remarkably higher than the latter (62.8%). The increasing time of thermal treatment increases the crystallinity of Pt(0) on carbon, accompanied by the increasing EAS areas, which corresponds to its enhanced electrocatalytic performance to methanol oxidation. PMID- 16706435 TI - Structure and catalytic properties of magnesia-supported copper salts of molybdovanadophosphoric acid. AB - The structure and stability of magnesia-supported copper salts of molybdovanadophosphoric acid (Cu(2)PMo(11)VO(40)) were characterized by different techniques. The catalyst was prepared in ethanol by impregnation because this solvent does not hurt texture of the water-sensitive MgO and Cu(2)PMo(11)VO(40). The Keggin-type structure compound may be degraded partially to form oligomerized polyoxometalate when supported on MgO. However, the oligomers can rebuild as the Keggin structure again after thermal treatment in air or during the reaction. Meanwhile, the V atoms migrate out of the Keggin structure to form a lacunary structure, as observed by Fourier transform IR spectroscopy. Moreover, the presence of Cu(2+) as a countercation showed an affirmative influence on the migration of V atoms, and the active sites derived from the lacunary species generated after release of V from the Keggin anion. The electron paramagnetic resonance data imply that V(5+) autoreduces to V(4+) in the fresh catalyst, and during the catalytic reaction a large number of V(4+) ions are produced, which enhance the formation of O(2-) vacancies around the metal atoms. These oxygen vacancies may also improve the reoxidation function of the catalyst. This behavior is correlated to higher catalytic properties of this catalyst. The oxidative dehydrogenation of hexanol to hexanal was studied over this catalyst. After reaction at 553.2 K for 50 h, catalytic properties did not decrease and exhibited higher selectivity (>96.0%) toward hexanal. PMID- 16706436 TI - Microscopic observation and in-situ Raman scattering studies on high-pressure phase transformations of Kr hydrate. AB - Direct observations through a microscope and in-situ Raman scattering measurements of synthesized single-crystalline Kr hydrate have been performed at pressures up to 5.2 GPa and 296 K. We have observed that the initial cubic structure II (sII) of Kr hydrate successively transforms to a cubic structure I (sI), a hexagonal structure, and an orthorhombic structure (sO) called "filled ice" at 0.45, 0.75, and 1.8 GPa, respectively. The sO phase exists at least up to 5.2 GPa. In addition to these transformations, we have also found the new phase behavior at 1.0 GPa, which is most likely caused by the change of cage occupancy of host water cages by guest Kr atoms without structural change. Raman scattering measurements for observed phases have shown that the lattice vibrational peak at around 130 cm(-1) disappears in the pressure region of sI, which enables us to distinguish the sI phase from sII and sH phases. PMID- 16706437 TI - Effect of SDS on the self-assembly behavior of the PEO-PPO-PEO triblock copolymer (EO)20(PO)70(EO)20. AB - The mixed micellar system comprising the poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide)-based triblock copolymer (EO)(20)(PO)(70)(EO)(20) (P123) and the anionic surfactant sodium dodecyl sulfate (SDS) has been investigated in aqueous media by small-angle neutron scattering (SANS) and viscosity measurements. The aggregation number of the copolymer in the micelles decreases upon addition of SDS, but a simultaneous enhancement in the degree of micellar hydration leads to a significant increase in the micellar volume fraction at a fixed copolymer concentration. This enhancement in the micellar hydration leads to a marked increase in the stability of the micellar gel phase until it is destroyed at very high SDS concentration. Mixed micellar systems with low and intermediate SDS concentrations form the micellar gel phase in much wider temperature and copolymer concentration ranges than the pure copolymer micellar solution. A comparison of the observed results with those for the copolymers (EO)(26)(PO)(40)(EO)(26) (P85) and (EO)(99)(PO)(70)(EO)(99) (F127) suggests that the composition of the copolymers plays a significant role in determining the influence of SDS on the gelation characteristics of the aqueous copolymer solutions. Copolymers with high PO/EO ratios show an enhancement in the stability of the gel phase, whereas copolymers with low PO/EO ratios show a deterioration of the same in the presence of SDS. PMID- 16706438 TI - Effect of graphitization on the wettability and electrical conductivity of CVD carbon nanotubes and films. AB - The use of carbon nanomaterials in various applications requires precise control of their surface and bulk properties. In this paper, we present a strategy for modifying the surface chemistry, wettability, and electrical conductivity of carbon tubes and films through annealing in a vacuum. Experiments were conducted with 60-300 nm nanotubes (nanopipes), produced by noncatalytic chemical vapor deposition (CVD) in a porous alumina template, and with thin films deposited by the same technique on a glassy carbon substrate having the same structure and chemistry of the CNTs. The surface of the as-produced CVD-carbon, treated with sodium hydroxide to remove the alumina template, is hydrophilic, and the bulk electrical conductivity is lower by a factor of 20 than that of fully graphitic multiwalled nanotubes (MWNT) or bulk graphite. The bulk electrical conductivity increases to the conductivity of graphite after annealing at 2000 degrees C in a high vacuum. The analysis of CNTs by transmission electron microscopy (TEM) and Raman spectroscopy shows the ordering of carbon accompanied by an exponential increase of the in-plane crystallite size, L(a), with increasing annealing temperature. Environmental scanning electron microscopy (ESEM) was used to study the interaction of CNT with water, and contact angle measurements performed using the sessile drop method on CVD-carbon films demonstrate that the contact angle increases nearly linearly with increasing annealing temperature. PMID- 16706439 TI - Comparative first-principles study of structural and optical properties of alkali metal azides. AB - A comparative first-principles study of the structural and optical properties of the alkali metal azides has been performed with density functional theory within the generalized gradient approximation. The crystal structures of the alkali azides compare well with experimental data. Their ionic character is manifested by the closeness of their internitrogen distances to the calculated N-N bond length for the free azide ion. An analysis of electronic structure, charge transfer, and bond order shows that the alkali azides are all wide-gap insulators and ionic compounds. The energy band and density of states for lithium azide and alpha-sodium azide are very similar, while these for potassium azide, alpha rubidium azide, and alpha-cesium azide are alike, but some modifications are observed with the increment of alkali metals' electropositivity. These changes are closely related to the differences of the crystal structures. The general shapes of the real and imaginary parts of the dielectric function, adsorption coefficient, and electron energy-loss spectra are quite similar. The peaks originate from the electron transitions from the alkali metal s and p states to the conduction band. Our calculated optical properties for the alkali azides are found to be in good agreement with available experimental data. The absorption spectra of the alkali azides show a number of absorption peaks, which are believed to be associated with different exciton states, in the fundamental absorption region. In general, the electron energy-loss spectra have two plasma frequencies. PMID- 16706440 TI - Thermally stable luminescent composites fabricated by confining rare earth complexes in the two-dimensional gallery of titania nanosheets and their photophysical properties. AB - Fluorescent composite materials of exfoliated titania nanosheets, Ti(0.91)O(2), and rare earth (RE) complexes, Eu(phen)(2)Cl(3).2H(2)O and Tb(phen)(2)Cl(3).2H(2)O (phen = 1,10-phenanthroline), were synthesized via flocculation between them. X-ray diffraction measurements and transmission electron microscopy observations confirmed a restacked lamellar structure for the composites, and elemental analysis revealed a high RE complex content of 15 wt %. The decomposition temperature of the complexes trapped in the composites was improved to 420 degrees C from 250 degrees C for the free form. The restacked composite composed of Ti(0.91)O(2) nanosheets and Eu(phen)(2) exhibited characteristic red emission from the complex, while the composite with Tb(phen)(2) gave featureless emission originated from the ligand. This phenomenon can be explained by a shift of triplet state level of the ligand after encapsulation in the host titania nanosheets. The quantum yield of europium complex in the composite was enhanced 1.6 times more than that of the pure complex. PMID- 16706441 TI - Phase separation in binary polymer/liquid crystal mixtures: network breaking and domain growth by coalescence-induced coalescence. AB - A small-angle light scattering (SALS) technique together with optical microscopy observation are used to investigate phase separation kinetics in films of low molecular weight thermotropic liquid crystal (4-cyano-4'-n-octyl-biphenyl, 8CB) with flexible polymer (polystyrene, PS). The growth of domains is studied as a function of time, film thickness, and film composition. The light scattering results are correlated with the images obtained by optical microscopy observation. In this paper, we study the breaking of a bicontinuous network of polymer in liquid crystal into droplets and their further growth via the coalescence-induced coalescence mechanism. The appearance of droplets in the system leads to a strong scattering at small wave vectors, while the bicontinuous network gives a peak at a nonzero wave vector. Superposition of these scattering intensities leads to the appearance of a second peak in the full scattering intensity signal, when the bicontinuous network starts to break up into disjointed elongated domains. Finally, both peaks merge into a single peak, which moves quickly toward zero wave vectors, indicating a complete transformation of elongated domains into spherical droplets of variable size. We found that the separation process does not depend on the size of the system. Irrespective of the sample thickness, the network breaks into fragments always at the same time after temperature quench. On the basis of morphological analysis, we found that the average size of the droplets which formed from the network grows with time, t, as t(alpha), alpha = 0.9 +/- 0.1, in the isotropic phase and in the nematic phase. PMID- 16706442 TI - Conformational features of an actuator containing calix[4]arene and thiophene: a molecular dynamics study. AB - Molecular dynamics simulations have been performed for poly(calix[4]arene bis(bithiophene)) in dichloromethane solution. This material responds to its electronic structure variations with significant conformational changes, producing contraction-expansion movements. Simulations have been performed for the three states of this molecular actuator (reduced, oxidized-nondeprotonated, and oxidized-deprotonated), a specific force-field being developed for each case. Results, which are fully consistent with previous ab initio quantum mechanical calculations on an isolated actuating unit, have revealed important findings about the dynamics of the system. Analyses of the flexibility/rigidity of the molecular chain with the state, the interaction of the polymer with the solvent molecules and the influence of environmental factors (as the viscosity of solvent, the counterions and the thermal agitation) on the dynamics have provided important insights to the actuation mechanism. PMID- 16706443 TI - Simulation and interpretation of 2D diffraction patterns from self-assembled nanostructured films at arbitrary angles of incidence: from grazing incidence (above the critical angle) to transmission perpendicular to the substrate. AB - A method to calculate the location of all Bragg diffraction peaks from nanostructured thin films for arbitrary angles of incidence from just above the critical angle to transmission perpendicular to the film is reported. At grazing angles, the positions are calculated using the distorted wave Born approximation (DWBA), whereas for larger angles where the diffracted beams are transmitted though the substrate, the Born approximation (BA) is used. This method has been incorporated into simulation code (called NANOCELL) and may be used to overlay simulated spot patterns directly onto two-dimensional (2D) grazing angle of incidence small-angle X-ray scattering (GISAXS) patterns and 2D SAXS patterns. The GISAXS simulations are limited to the case where the angle of incidence is greater than the critical angle (alpha(i) > alpha(c)) and the diffraction occurs above the critical angle (alpha(f) > alpha(c)). For cases of surfactant self assembled films, the limitations are not restrictive because, typically, the critical angle is around 0.2 degrees but the largest d spacings occur around 0.8 degrees 2theta. For these materials, one finds that the DWBA predicts that the spot positions from the transmitted main beam deviate only slightly from the BA and only for diffraction peaks close the critical angle. Additional diffraction peaks from the reflected main beam are observed in GISAXS geometry but are much less intense. Using these simulations, 2D spot patterns may be used to identify space group, identify the orientation, and quantitatively fit the lattice constants for SAXS data from any angle of incidence. Characteristic patterns for 2D GISAXS and 2D low-angle transmission SAXS patterns are generated for the most common thin film structures, and as a result, GISAXS and SAXS patterns that were previously difficult to interpret are now relatively straightforward. The simulation code (NANOCELL) is written in Mathematica and is available from the author upon request. PMID- 16706444 TI - Dithiocarbamate anchoring in molecular wire junctions: a first principles study. AB - Recent experimental realization [J. Am. Chem. Soc., 127 (2005) 7328] of various dithiocarbamate self-assembly on gold surface opens the possibility for use of dithiocarbamate linkers to anchor molecular wires to gold electrodes. In this paper, we explore this hypothesis computationally. We computed the electron transport properties of 4,4'-bipyridine (BP), 4,4'-bipyridinium-1,1' bis(carbodithioate) (BPBC), 4-(4'-pyridyl)-peridium-1-carbodithioate (BPC) molecule junctions based on the density functional theory and nonequilibrium Green's functions. We demonstrated that the stronger molecule-electrode coupling associated with the conjugated dithiocarbamate linker broadens transmission resonances near the Fermi energy. The broadening effect along with the extension of the pi conjugation from the molecule to the gold electrodes lead to enhanced electrical conductance for BPBC molecule. The conductance enhancement factor is as large as 25 at applied voltage bias 1.0 V. Rectification behavior is predicted for BPC molecular wire junction, which has the asymmetric anchoring groups. PMID- 16706445 TI - X-ray absorption spectroscopy of titanium oxide by time dependent density functional calculations. AB - The potentiality of the time dependent density functional theory (TDDFT) for the description of core excitation spectra (XAS) in transition metal oxides is analyzed, considering the rutile form of TiO(2) as a test case. Cluster models are adopted to mimic the bulk, embedded within an array of point charges to simulate the Madelung potential. All of the edges, titanium and oxygen K and titanium L edges, are considered, and the TDDFT results are compared with the experimental data in order to assess the performance of the theoretical approach in dealing with this complex class of compounds. Satisfactory results have been obtained for the Ti and O K edges, while in the case of the Ti L edge some discrepancies with the experiment are still present. The configuration mixing explicitly included in the TDDFT model strongly influences the distribution of the 2p metal oscillator strength. The origin of the spectral features is investigated with the help of the partial density of the virtual states (PDOS) calculated for each core hole considered, which can be qualitatively compared with the theoretical spectra calculated in the Kohn-Sham one-electron approach. PMID- 16706447 TI - In situ extended X-ray absorption fine structure study during selective alcohol oxidation over Pd/Al2O3 in supercritical carbon dioxide. AB - High-pressure in situ X-ray absorption near-edge structure (XANES) and extended X ray absorption fine structure (EXAFS) data are reported during the selective oxidation of benzyl alcohol to benzaldehyde in supercritical carbon dioxide over a Pd/Al(2)O(3) catalyst (shell impregnated). For this purpose, a continuous-flow system with a spectroscopic cell suitable for in situ X-ray absorption studies on heterogeneous catalysts up to 200 degrees C and 200 bar has been developed. Due to the high contribution of the dense fluid to the overall X-ray absorption, high stability of the process pressure is mandatory, particularly when recording EXAFS spectra. According to EXAFS and XANES results, the palladium particles were fully reduced after exposure to benzyl alcohol in scCO(2). In contrast to Pd-catalyzed liquid-phase oxidation, a higher oxygen tolerance of the catalyst was observed. Palladium was partially oxidized on the surface under typical reaction conditions (0.9 mol % benzyl alcohol/0.5 mol % O(2) in carbon dioxide), which gradually increased when the concentration of oxygen in the feed was raised. Both XANES and EXAFS data uncovered that palladium is mainly oxidized on the surface or within the outermost layers. These results are in accordance with simulations of the XANES data using the FEFF8.20 code (program for ab initio calculations on multiple scattering XAS) and EXAFS data fitting/simulation. PMID- 16706446 TI - Electronic excitation of polyfluorenes: a theoretical study. AB - We present systematic, theoretical investigations on structure-property correlations in polyfluorenes (PFs) derived mainly from the chain morphology, oligomer length, and chemical substitutent. Both the vertical absorptions and the vibrational contributions to electronic absorption and fluorescence spectra have been calculated. The effect of temperature on the nature of photoexcitations of PFs has been demonstrated. It is found that the vibronic (electronic and vibrational) structures of PFs are morphology-dependent. beta-phase oligofluorenes (beta-(FL)(n)) and ladder-type poly(p-phenylene) (LPPP) oligomers show a red shift compared to the spectra of alpha-(FL)(n). The asymmetry of the absorption and fluorescence spectra in alpha-(FL)(n) and the fluorenone (FLO) defect oligofluorenes alpha-(FL)(n)(-)(m)(FLO)(m) is significantly more pronounced than that in planarized beta-(FL)(n) and LPPP oligomers. By properly taking into account the anharmonic torsion potentials resulting from the strong electronic and nuclear coupling in the oligofluorenes, we have reasonably reproduced the experimentally observed spectroscopic features. The low-energy on chain chemical defect sites such as FLO units act as charge-trapping sites for singlet excitations, are the predominantly lighting-emitting species, and thus alter the blue light-emitting properties of PFs whereas the blue-light-emitting properties of PFs are hardly influenced by the hole-transporting molecules. The optical properties of PFs have been predicted by the finite-size calculations. Energy gaps of PFs are estimated by extrapolations from excitation energies of oligofluorenes up to 21 FL units. PMID- 16706448 TI - Gate-controlled rectifying behavior in C70@SWNT networks. AB - We report the gate-controlled rectification behavior in C(70)@SWNT networks at room temperature in air. The electrical transport characteristics can be fitted well with the conventional Schottky diode model. The origin of the rectifying behavior in fullerene peapod networks device is qualitatively discussed. This paper demonstrates a strategy for diode fabrication based on peapod networks. PMID- 16706449 TI - Divide and protect: capping gold nanoclusters with molecular gold-thiolate rings. AB - Density functional theory calculations are used to explore phosphine- and thiolate-protected gold nanoclusters, namely, Au(39)(PH(3))(14)Cl(6) and Au(38)(SCH(3))(24). For Au(38)(SCH(3))(24), a novel structural motif is predicted, consisting of ringlike (AuSCH(3))(4) units protecting a central Au(14) core. The calculated optical spectrum of this species features a large optical gap (about 1.5 eV) and a prominently peaked structure, correlating with experimental findings of "molecular-like spectra" of thiolate-protected 1.1 nm gold nanoparticles. Ligand-ligand interactions and steric effects in the ligand shell are suggested as possible driving forces toward an ordered gold core structure. A novel mechanism for ligand-exchange reactions on gold clusters is proposed. PMID- 16706451 TI - Phenyl species formation and preferential hydrogen abstraction in the decomposition of chemisorbed benzoate on Cu(110). AB - The adsorption and decomposition of benzoic acid on the Cu(110) surface has been investigated using temperature-programmed reaction (TPR) spectroscopy and scanning tunneling microscopy (STM). The benzoate species is found to exist in two conformations--a phase containing upright species at monolayer saturation and a phase containing many lying-down species at lower coverages. Thermal decomposition begins to occur near 500 K, yielding benzene and CO(2). It is found that phenyl species, generated preferentially from the lying-down benzoate species, efficiently abstract H atoms from undecomposed benzoate species to produce benzene in a rate-controlling process with an activation energy of about 29 kcal/mol. Using deuterium-atom substitution at the 4-C position on the benzoate ring it is found that the hydrogen-abstraction reaction is selective for 2,3 and 5,6 C-H bonds. This observation indicates that the mobile phenyl species is surface bound and preferentially attacks C-H bonds which are nearest the Cu surface and bind the benzoate species as either an upright species or a tilted species. PMID- 16706450 TI - Pt and Ru X-ray absorption spectroscopy of PtRu anode catalysts in operating direct methanol fuel cells. AB - In situ X-ray absorption spectroscopy, ex situ X-ray fluorescence, and X-ray powder diffraction enabled detailed core analysis of phase segregated nanostructured PtRu anode catalysts in an operating direct methanol fuel cell (DMFC). No change in the core structures of the phase segregated catalyst was observed as the potential traversed the current onset potential of the DMFC. The methodology was exemplified using a Johnson Matthey unsupported PtRu (1:1) anode catalyst incorporated into a DMFC membrane electrode assembly. During DMFC operation the catalyst is essentially metallic with half of the Ru incorporated into a face-centered cubic (FCC) Pt alloy lattice and the remaining half in an amorphous phase. The extended X-ray absorption fine structure (EXAFS) analysis suggests that the FCC lattice is not fully disordered. The EXAFS indicates that the Ru-O bond lengths were significantly shorter than those reported for Ru-O of ruthenium oxides, suggesting that the phases in which the Ru resides in the catalysts are not similar to oxides. PMID- 16706452 TI - Kinetic isotope effect for H2 and D2 quantum molecular sieving in adsorption/desorption on porous carbon materials. AB - Adsorption and desorption of H(2) and D(2) from porous carbon materials, such as activated carbon at 77 K, are usually fully reversible with very rapid adsorption/desorption kinetics. The adsorption and desorption of H(2) and D(2) at 77 K on a carbon molecular sieve (Takeda 3A), where the kinetic selectivity was incorporated by carbon deposition, and a carbon, where the pore structure was modified by thermal annealing to give similar pore structure characteristics to the carbon molecular sieve substrate, were studied. The D(2) adsorption and desorption kinetics were significantly faster (up to x1.9) than the corresponding H(2) kinetics for specific pressure increments/decrements. This represents the first experimental observation of kinetic isotope quantum molecular sieving in porous materials due to the larger zero-point energy for the lighter H(2), resulting in slower adsorption/desorption kinetics compared with the heavier D(2). The results are discussed in terms of the adsorption mechanism. PMID- 16706453 TI - Theoretical and spectroscopic study of the effect of ring substitution on the adsorption of anisole on platinum. AB - The adsorption of anisole, 3,5-dimethylanisole, and 3,5-bis-(trifluoromethyl) anisole on Pt(111) was studied theoretically and compared to the adsorption of benzene using relativistically corrected density functional theory. A cluster of 31 platinum atoms was used to simulate the surface. The three anisoles were found to be less strongly adsorbed than the parent molecule benzene, 3,5-bis (trifluoromethyl)-anisole showing weakest adsorption, with an adsorption energy of only one-third that of benzene. The theoretical study was complemented by in situ ATR-IR spectroscopy of the adsorption of the anisole derivatives on a polycrystalline Pt film. The spectroscopic study indicated that the adsorption strength of the anisoles follows the same order as predicted by the calculations. In addition, catalytic hydrogenation tests showed that the propensity to aromatic ring hydrogenation can also be correlated to the mode and strength of adsorption of the anisoles. The degree of saturation followed the same order as the adsorption strength found by the calculations and indicated by spectroscopy. Although 3,5-dimethyl substitution on anisole resulted in only a partial loss of adsorption energy and reactivity toward ring hydrogenation as compared to anisole, the substitution by CF(3) groups led to a large loss of adsorption energy and complete loss of reactivity toward aromatic ring saturation. Along with the study of the substituent effect on the adsorption of aromatic molecules, the correlation between adsorption and propensity to saturation of aromatic substrates could be corroborated. PMID- 16706454 TI - Kinetics of the reduction of the Rh(111) surface oxide: linking spectroscopy and atomic-scale information. AB - The reduction of the surface oxide on Rh(111) by H(2) was observed in situ by scanning tunneling microscopy (STM) and high-resolution core level spectroscopy (HRCLS). At room temperature, H(2) does not adsorb on the oxide, only in reduced areas. Reduction starts in very few sites, almost exclusively in stepped areas. One can also initiate the reduction process by deliberately creating defects with the STM tip allowing us to examine the reduction kinetics in detail. Depending on the size of the reduced area and the hydrogen pressure, two growth regimes were found. At low H(2) pressures or small reduced areas, the reduction rate is limited by hydrogen adsorption on the reduced area. For large reduced areas, the reduction rate is limited by the processes at the border of the reduced area. Since a near-random distribution of the reduction nuclei was found and the reduction process at defects starts at a random time, one can use Johnson-Mehl Avrami-Kolmogoroff (JMAK) theory to describe the process of reduction. The microscopic data from STM agree well with spatially averaged data from HRCLS measurements. PMID- 16706455 TI - CO2 reforming of CH4 on Ni(111): a density functional theory calculation. AB - CO(2) reforming of CH(4) on Ni(111) was investigated by using density functional theory. On the basis of thermodynamic analyses, the first step is CH(4) sequential dissociation into surface CH (CH(4) --> CH(3) --> CH(2) --> CH) and hydrogen, and CO(2) dissociation into surface CO and O (CO(2) --> CO + O). The second step is CH oxygenation into CHO (CH + O --> CHO), which is more favored than dissociation into C and hydrogen (CH --> C + H). The third step is the dissociation of CHO into surface CO and H (CHO --> CO + H). This can explain the enhanced selectivity toward the formation of CO and H(2) on Ni catalysts. It is found that surface carbon formation by the Bouduard back reaction (2CO = C((ads)) + CO(2)) is more favored than by CH(4) sequential dehydrogenation. The major problem of CO(2) reforming of CH(4) is the very strong CO adsorption on Ni(111), which results in the accumulation of CO on the surface and hinders the subsequent reactions and promotes carbon deposition. Therefore, promoting CO desorption should maintain the reactivity and stability of Ni catalysts. The computed energy barriers of the most favorable elementary reaction identify the CH(4) activation into CH(3) and H as the rate-determining step of CO(2) reforming of CH(4) on Ni(111), in agreement with the isotopic experimental results. PMID- 16706456 TI - Phase segregation in cerium-lanthanum solid solutions. AB - Electron energy-loss spectroscopy (EELS) in combination with scanning transmission electron microscopy (STEM) reveals that the La enrichment at the surface of cerium-lanthanum solid solutions is an averaged effect and that segregation occurs in a mixed oxide phase. This separation occurs within a crystalline particle, where the dopant-rich phase is located at the surface of the dopant-deficient phase. The limiting structure appears to be a solid solution with a La fraction of x = 0.6 in the bulk and x = 0.75 at the surface. Up to a La fraction of 0.6, this phase will coexist with a lanthanum-type structure in different proportions depending on the dopant amount. STEM-EELS appears to be a powerful technique to clarify the existence of a multiphase system, and it shows that XRF, XPS, and XRD measure averaged results and do not show the phase complexity of the solids. PMID- 16706457 TI - Conducting-probe AFM nanoscale joule heating yields charge-density-wave transition detection. AB - Several layered transition-metal dichalcogenides are studied using conducting probe AFM aiming to investigate the probe-mediated thermal processes likely to arise in the probe-substrate vicinity due to the high-current densities involved. A signature of local heating is found in the shape of current-potential (i/V) curves. The latter allows straightforward detection of a charge-density-wave (CDW) transition for 1T-TaSe(2) samples exhibiting it above room temperature. This is an illustration of a new use of conducting probe AFM to investigate solid state bulk characteristics owing to a distinctive nanoscale Joule heating. PMID- 16706458 TI - Defect chemistry, surface structures, and lithium insertion in anatase TiO2. AB - Atomistic simulation techniques are used to investigate the defect properties of anatase TiO(2) and Li(x)TiO(2) both in the bulk and at the surfaces. Interatomic potential parameters are derived that reproduce the lattice constants of anatase, and the energies of bulk defects and surface structures are calculated. Reduction of anatase involving interstitial Ti is found to be the most favorable defect reaction in the bulk, with a lower energy than either Frenkel or Schottky reactions. The binding energies of selected defect clusters are also presented: for the Ti(3+)-Li(+) defect cluster, the binding energy is found to be approximately 0.5 eV, suggesting that intercalated Li ions stabilize conduction band electrons. The Li ion migration path is found to run between octahedral sites, with an activation energy of 0.45-0.65 eV for mole fractions of lithium in Li(x)TiO(2) of x < or = 0.1. The calculated surface energies are used to predict the crystal morphology, which is found to be a truncated bipyramid in which only the (101) and (001) surfaces are expressed, in accord with the available microscopy data. Calculations of defect energies at the (101) surface suggest that single Ti(3+) defects and neutral Ti(3+)-Li(+) pairs tend to segregate to the surface. PMID- 16706459 TI - Determination of band curvatures by angle-resolved two-photon photoemission in thin films of C60 on Ag(111). AB - The thickness-dependent interfacial band structure was determined for thin films of C(60) on Ag(111) by angle-resolved two-photon photoemission spectroscopy. Dispersions of molecular-orbital derived bands (HOMO, LUMO+1, and LUMO+2) were acquired, and limits were placed on their possible effective masses. A group theoretic approach is also incorporated to further understand the properties of these states. The HOMO, LUMO+1, and LUMO+2 bands possess (best-fit) effective masses of -7 m(e), -7 m(e), and -12 m(e), respectively. These values are consistent with theoretical calculations, averaged over the closely spaced subbands for each state, and provide practical limits on the effective fundamental charge-transport properties of C(60) films. PMID- 16706460 TI - Simultaneous nanoindentation and electron tunneling through alkanethiol self assembled monolayers. AB - Electrical tunnel junctions consisting of alkanethiol molecules self-assembled on Au-coated Si substrates and contacted with Au-coated atomic force microscopy tips were characterized under varying junction loads in a conducting-probe atomic force microscopy configuration. Junction load was cycled in the fashion of a standard nanoindentation experiment; however, junction conductance rather than probe depth was measured directly. The junction conductance data have been analyzed with typical contact mechanics (Derjaguin-Muller-Toporov) and tunneling equations to extract the monolayer modulus (approximately 50 GPa), the contact transmission (approximately 2 x 10(-6)), contact area, and probe depth as a function of load. The monolayers are shown to undergo significant plastic deformation under compression, yielding indentations approximately 7 Angstroms deep for maximum junction loads of approximately 50 nN. Comparison of mechanical properties for different chain lengths was also performed. The film modulus decreased with the number of carbons in the molecular chain for shorter-chain films. This trend abruptly reversed once 12 carbons were present along the backbone. PMID- 16706461 TI - Structure sensitivity of methanol electrooxidation pathways on platinum: an on line electrochemical mass spectrometry study. AB - By monitoring the mass fractions of CO(2) (m/z 44) and methylformate (m/z 60, formed from CH(3)OH + HCOOH) with on-line electrochemical mass spectrometry (OLEMS), the selectivity and structure sensitivity of the methanol oxidation pathways were investigated on the basal planes--Pt(111), Pt(110), and Pt(100)- and the stepped Pt electrodes--Pt(554) and Pt(553)--in sulfuric and perchloric acid electrolytes. The maximum reactivity of the MeOH oxidation reaction on Pt(111), Pt(110), and Pt(100) increases in the order Pt(111) < Pt(110) < Pt(100). Mass spectrometry results indicate that the direct oxidation pathway through soluble intermediates plays a pronounced role on Pt(110) and Pt(111), while, on Pt(100), the indirect pathway through adsorbed carbon monoxide is predominant. In 0.5 M H(2)SO(4), introducing steps in the (111) plane increases the total reaction rate, while the relative importance of the direct pathway decreases considerably. In 0.5 M HClO(4), however, introducing steps increases both the total reaction rate and the selectivity toward the direct oxidation pathway. Anion (sulfate) adsorption on (111) leads to a more prominent role of the direct pathway, but, on all the other surfaces, (bi)sulfate seems to block the formation of soluble intermediates. For both electrolytes, increasing the step density results in more methylformate being formed relative to the amount of CO(2) detected, indicating that the [110] steps themselves catalyze the direct oxidation pathway. A detailed reaction scheme for the methanol oxidation mechanism is suggested based on the literature and the results obtained here. PMID- 16706462 TI - Breakdown of the Gouy-Chapman model for highly charged Langmuir monolayers: counterion size effect. AB - Deviations from the classic Gouy-Chapman (GC) model due to the finite size of hydrated counterions were tested for negatively charged Langmuir monolayers with different surface charge densities. Monolayers with the largest charge density (>0.6 C.m(-2)) show an increase of the surface potential for a series of alkali metal cations from Li(+) to Cs(+) by 200-250 mV. The increase is similar for different monolayers and suggests that this effect is independent of the particular type of headgroup. The magnitude of variation is comparable with model estimations of the electrical double layer (EDL) potential implying that the deviation from the GC model is drastic. Deviations from the GC model rapidly vanish with decreasing monolayer charge density and become hardly observable below 0.3 C.m(-2). For monolayers with a high charge density on subphases containing different sized counterions, preferential participation of the smallest ions in the EDL should be favorable in terms of electrostatic free energy because of packing density limitations. This effect was demonstrated for behenyl sulfate (BS) monolayers (0.64 C.m(-2)) with the X-ray reflectivity technique. For the Cs(+)-Li(+) system, the fraction of Cs(+) in the EDL is 50-60% compared with only 10% of Cs(+) in the subphase. Providing high surface charge density, a small univalent Cs(+) is capable to compete even with a bulky divalent Mg(2+). For equal concentrations of Cs(+) and Mg(2+) in the subphase, the Cs(+)/Mg(2+) ratio in EDL of BS monolayer is 1.3 to 2.0 (in contrast to 0.04, predicted by the GC model). All experimental results of this study are described in terms of packing density limitations for hydrated counterions in the EDL. PMID- 16706463 TI - Physical properties of gamma alumina surface hydroxyls revisited through a large scale periodic quantum-chemistry approach. AB - We have studied surface hydroxyls adsorbed onto (001), (011), and (111) gamma alumina surfaces using a quantum-chemistry approach in order to compare with empirical models proposed in the literature. Local electronic structures and geometries in the low OH coverage limit have been evaluated for both ideal and relaxed surfaces with the help of a large scale periodic quantum-chemical code. Hydroxyl groups are adsorbed onto surfaces, and a study of their local electronic structure, vibrational frequencies, charges, and adsorption energies is performed and analyzed as a function of their adsorption site geometry. Our results show that, even on ideal (nonrelaxed) surfaces, OH local environments are more complicated than those stated by empirical models and strongly influence the hydroxyl stretching vibrational mode. Large scale simulation shows that disorder takes place even at 0 K, and the analysis of the vibrational frequencies leads to a revision of Knozinger's empirical model. Cationic vacancies in the first surface layers have also been taken into account; they have a significant influence on the surface atomic and electronic structures, modifying the physical properties of adsorbed OH entities. This work emphasizes the necessity to perform an electronic structure calculation to better understand adsorbed OH properties on gamma alumina surfaces and reveals the difficulty to make a one-to-one correspondence between OH stretching frequencies and their other physical properties. Finally, we show that these results agree with some available experimental studies. PMID- 16706464 TI - In situ sum frequency generation vibrational spectroscopy observation of a reactive surface intermediate during high-pressure benzene hydrogenation. AB - Sum frequency generation surface vibrational spectroscopy and kinetic measurements using gas chromatography have been used to identify a reactive surface intermediate in situ during hydrogenation of benzene on a Pt(111) single crystal surface at Torr pressures. Upon adsorption at 310 K, both chemisorbed and physisorbed benzene coexist on the surface, a result which has not previously been observed. Kinetic measurements show a linear compensation effect for the production of both cyclohexane and cyclohexene. From these data the isokinetic temperature was identified and correlated to the chemisorbed benzene species, which were probed by means of vibrational spectroscopy. Additionally, chemisorbed benzene was determined to be a reactive intermediate, which is critical for hydrogenation. PMID- 16706465 TI - Influence of chlorine substitution on the self-assembly of zinc phthalocyanine. AB - The adsorption and ordering of zinc phthalocyanine (ZnPc) and octachloro zinc phthalocyanine (ZnPcCl(8)) on an Ag(111) surface is studied in situ by scanning tunneling microscopy under ultrahigh vacuum. Two-dimensional self-assembled supramolecular domains are observed for these two molecules. We show how substituting chlorine atoms for half of the peripheral hydrogen atoms on ZnPc influences the self-assembly mechanisms. While intermolecular interactions are dominated by van der Waals forces in ZnPc molecular networks, ZnPcCl(8) molecular packing undergoes a sequential phase evolution driven by the creation of C-Cl...H C hydrogen bonds between adjacent molecules. At the end of this evolution, the final molecular assembly involves all possible hydrogen bonds. Our study also reveals the influence of molecule-substrate interactions through the presence of fault lines generating a stripe structure in the molecular film. PMID- 16706466 TI - Adsorption dynamics and electrochemical and photophysical properties of thiolated ruthenium 2,2'-bipyridine monolayers. AB - A new complex [Ru(bpy)(2)(bpySH)](PF(6))(2), RuBpySH, has been prepared bearing two anchoring groups for surface attachment, where bpy is 2,2'-bipyridyl and bpySH is 5,5'-bis(mercaptomethyl)-2,2'-bipyridine. Monolayers of RuBpySH have been formed on micro and macro platinum electrodes by spontaneous adsorption from micromolar solutions of the complex in 50:50 v/v water/acetone. The monolayers can be reversibly switched between the Ru(2+) and the Ru(3+) forms. Cyclic voltammetry is well-defined with a peak-to-peak splitting of 30 +/- 5 mV and a full width at half-maximum of 110 +/- 10 mV being observed for scan rates up to 5 V s(-1) where the supporting electrolyte is 0.1 M tetrabutylammonium tetrafluoroborate in acetonitrile. Adsorption is irreversible in this system, and the saturation coverage obtained is 8.1 +/- 0.4 x 10(-11) mol cm(-2) when the complex concentration in the deposition solution is between 10 microM and 1.0 mM. The dynamics of adsorption depend markedly on the bulk concentration and are described in terms of irreversible adsorption. Dry monolayers display luminescence properties similar to those of powder samples of the complex, indicating that the monolayer has characteristics of the solid-state sample rather than the solution sample of the complex. Significantly, efficient electrochemiluminescence is generated using tripropylamine as the coreactant. The rate of electron transfer across the electrode/monolayer interface has been probed using high scan rate cyclic voltammetry. The standard heterogeneous electron-transfer rate constant, k degrees , is 0.9 +/- 0.1 x 10(4) s(-1), and there is weak adsorbate-electrode electronic communication. PMID- 16706467 TI - Adsorption of cationic lipid bilayer onto flat silicon wafers: effect of ion nature and concentration. AB - The effect of monovalent salt nature and concentration over a range of low ionic strengths (0-10 mM LiCl, NaCl, KCl, or CsCl) and at two different pH values (6.3 and 10.0) on adsorption of dioctadecyldimethylammonium bromide (DODAB) bilayer fragments (BF) onto flat SiO(2) surfaces was systematically evaluated by means of in situ ellipsometry. High-affinity adsorption isotherms fitted by the Langmuir model indicated that adsorption maxima were consistent with bilayer deposition only around 10 mM monovalent salt at both pH values. In pure water, the mean thickness of the DODAB adsorbed layer was close to zero with bilayer deposition taking place only around 10 mM ionic strength. In the presence of 10 mM CsCl or LiCl, the highest and the lowest affinity constants for DODAB adsorption onto SiO(2) were, respectively, obtained consistently with the expected facility of cation exchange at the surface required for DODAB adsorption. The cation more tightly bound to the solid surface should be Li(+), which would present the largest resistance to displacement by the DODAB cation, whereas the less tightly bound cation should be Cs(+) due to its largest ionic radius and lowest charge density. In other words, DODAB adsorption proceeds in accordance with charge density on the solid surface, which depends on the nature and concentration of bound counterions as well as DODAB cation ability to displace them. AFM images show a very smooth DODAB film adsorbed onto the surface in situ with a large frequency of BF auto-association from their edges. The present results for flat surfaces entirely agree with previous data from our group for DODAB adsorption onto silica particles. PMID- 16706468 TI - On the clarification of the IR stretching vibration assignment of adsorbed N2 on Rh0 and Rhdelta+ surface atoms of supported Rh crystallites. AB - The low-temperature adsorption of N(2) on Rh/SiO(2) samples of various particle size distributions was followed by FTIR. The addition of O(2) pulses on Rh(0) surfaces saturated with chemisorbed N(2) allowed us to reassign stretching frequencies attributed originally to N(2)-Rh(0) to N(2)-Rh(delta+). The formation of the latter oxidized Rh species is assumed to be induced by an electron withdrawal from adsorbed oxygen species on Rh surface centers neighboring those onto which N(2) species are chemisorbed. The present work, thus, enables us to delimit ranges of frequencies for which the adsorption of N(2) can be considered to occur on either Rh(0) or Rh(delta+) centers for nu(N2) lower or higher than 2243 cm(-1), respectively. The N(2)-FTIR experiments performed on the studied catalysts also suggest a lattice plane selectivity for N(2) adsorption on metallic Rh planes of different natures which, to our knowledge, has not been reported yet for Rh. PMID- 16706469 TI - Theoretical analysis of a novel electrical field assisted membrane module comprising an array of microchannel units. AB - A novel electrical field assisted membrane module consisting of an array of microchannel units, each microchannel unit comprised of a cylindrical pore and a charged ion-selective membrane layer, is analyzed theoretically. The governing equations for the flow and the electrical fields are solved analytically under the Debye-Huckel condition and the influences of the key parameters on the flow behavior of the system under consideration are investigated through numerical simulation. We show that for a fixed microchannel radius, the volumetric flow rate through a microchannel unit has a maximal value as the radius of the cylindrical pore varies. This maximum is independent of both the strength of the applied field and the density of the fixed charges in the membrane layer, but varies with the permittivity of the membrane layer. PMID- 16706470 TI - Single-sphere model for absorption spectrum of interfacial molecules with application to predictions of orientational angles. AB - In this work, a novel expression for nonequilibrium free energy is introduced within the framework of the continuum model. A simple and analytical model for the solvent effect of absorption spectrum of an interfacial molecule is deduced. This model overcomes some lacks existing in the previous theoretical treatments. Associated with the experimental data of N,N-diethyl-p-nitroaniline and 4-(2,4,6 triphenylpyridinium)-2,6-diphenylphenoxide, the present model gives reasonable predication of orientational angles for different interfaces. PMID- 16706471 TI - Reorganization asymmetry of electron transfer in ferroelectric media and principles of artificial photosynthesis. AB - This study considers electronic transitions within donor-acceptor complexes dissolved in media with macroscopic polarization. The change of the polarizability of the donor-acceptor complex in the course of electronic transition couples to the reaction field of the polar environment and the electric field created by the macroscopic polarization. An analytical theory developed to describe this situation predicts a significant asymmetry of the reorganization energy between charge separation and charge recombination transitions. This result is proved by Monte Carlo simulations of a model polarizable diatomic dissolved in a ferroelectric fluid of soft dipolar spheres. The ratio of the reorganization energies for the forward and backward reactions up to a factor of 25 is obtained in the simulations. This result, as well as the effect of the macroscopic electric field, is discussed in application to the design of efficient photosynthetic devices. PMID- 16706472 TI - Solid-solid phase transformation via internal stress-induced virtual melting, significantly below the melting temperature. Application to HMX energetic crystal. AB - We theoretically predict a new phenomenon, namely, that a solid-solid phase transformation (PT) with a large transformation strain can occur via internal stress-induced virtual melting along the interface at temperatures significantly (more than 100 K) below the melting temperature. We show that the energy of elastic stresses, induced by transformation strain, increases the driving force for melting and reduces the melting temperature. Immediately after melting, stresses relax and the unstable melt solidifies. Fast solidification in a thin layer leads to nanoscale cracking which does not affect the thermodynamics or kinetics of the solid-solid transformation. Thus, virtual melting represents a new mechanism of solid-solid PT, stress relaxation, and loss of coherence at a moving solid-solid interface. It also removes the athermal interface friction and deletes the thermomechanical memory of preceding cycles of the direct-reverse transformation. It is also found that nonhydrostatic compressive internal stresses promote melting in contrast to hydrostatic pressure. Sixteen theoretical predictions are in qualitative and quantitative agreement with experiments conducted on the PTs in the energetic crystal HMX. In particular, (a) the energy of internal stresses is sufficient to reduce the melting temperature from 551 to 430 K for the delta phase during the beta --> delta PT and from 520 to 400 K for the beta phase during the delta --> beta PT; (b) predicted activation energies for direct and reverse PTs coincide with corresponding melting energies of the beta and delta phases and with the experimental values; (c) the temperature dependence of the rate constant is determined by the heat of fusion, for both direct and reverse PTs; results b and c are obtained both for overall kinetics and for interface propagation; (d) considerable nanocracking, homogeneously distributed in the transformed material, accompanies the PT, as predicted by theory; (e) the nanocracking does not change the PT thermodynamics or kinetics appreciably for the first and the second PT beta <--> delta cycles, as predicted by theory; (f) beta <--> delta PTs start at a very small driving force (in contrast to all known solid-solid transformations with large transformation strain), that is, elastic energy and athermal interface friction must be negligible; (g) beta --> alpha and alpha --> beta PTs, which are thermodynamically possible in the temperature range 382.4 < theta < 430 K and below 382.4 K, respectively, do not occur. PMID- 16706473 TI - Solvation dynamics of C153 in supercritical fluoroform: a simulation study based on two-site and five-site models of the solvent. AB - Molecular dynamics (MD) simulations of a probe solute (coumarin C153) in supercritical fluoroform are used to study time-dependent solute-solvent interactions. We study the dynamics of solvent reorganization in response to electronic excitation of C153 at a temperature of 1.03 T(c) (the critical temperature) and a series of densities above and below the critical density. Simulations of a two-site and five-site models of fluoroform are presented and compared. The time-dependent solvation response after solute electronic excitation is studied in the two cases, and the five-site results present an earlier onset of exponential decay that is closer to what is expected to be the experimental response. This is confirmed by comparison to experiment. In addition to obtaining the solvation response from nonequilibrium MD trajectories, approximate solvation responses were obtained from equilibrium time correlations of the fluctuations in the solvation energy change in the presence of ground- and excited-state solutes. For the five-site model, the equilibrium excited-state response shows stronger density dependence than the ground-state one. The nonequilibrium response appears to have an intermediate decay rate between the two equilibrium functions. The solute-partial-charge-solvent-induced-dipole interaction was also taken into account by means of a perturbative approach, which improved the agreement with experimental measurements available at densities corresponding to 1.4-1.6 rho(c) (where rho(c) the critical density). From the comparison between the two models, it is possible to conclude that an atomistic description is necessary for correctly representing the portion of solvation dynamics that is related to reorientation. This consideration is supported by providing results for orientational time correlation functions and by comparing the correlation times with the experimental ones. PMID- 16706474 TI - Comparison of coupled motions in Escherichia coli and Bacillus subtilis dihydrofolate reductase. AB - Hybrid quantum/classical molecular dynamics simulations are used to compare the role of protein motion in the hydride transfer reaction catalyzed by Escherichia coli and Bacillus subtilis dihydrofolate reductase (DHFR). These two enzymes have 44% sequence identity, and the experimentally determined structures and hydride transfer rates are similar. The simulations indicate that the tertiary structures of both enzymes evolve in a similar manner during the hydride transfer reaction. In both enzymes, the donor-acceptor distance decreases to approximately 2.7 Angstroms at the transition state configurations to enable hydride transfer. Zero point energy and hydrogen tunneling effects are found to be significant for both enzymes. Covariance and rank correlation analyses of motions throughout the protein and ligands illustrate that E. coli and B. subtilis DHFR exhibit both similarities and differences in the equilibrium fluctuations and the conformational changes along the collective reaction coordinate for hydride transfer. A common set of residues that play a significant role in the network of coupled motions leading to configurations conducive to hydride transfer for both E. coli and B. subtilis DHFR was identified. These results suggest a balance between conservation and flexibility in the thermal motions and conformational changes during hydride transfer. Homologous protein structures, in conjunction with conformational sampling, enable enzymes with different sequences to catalyze the same hydride transfer reaction with similar efficiency. PMID- 16706475 TI - Investigations into the analysis and modeling of the cytochrome P450 cycle. AB - The main focus of our research is to explore the fundamental dynamics of the mechanism of the cytochrome P450 (CYP450) cycle. For this purpose we propose a system-theoretical approach, a time-dependent metabolic control analysis (tdMCA), to the analysis and quantitative modeling of the CYP450 catalytic pathway. This provides theoretical enlightenment for us to assess the transient response of the system to perturbations. In addition, the robustness of the cycle has also been observed, where perturbations elicit very weak responses and the system quickly recovers to the steady state (in an average of 10(-5) s). The tdMCA also shows that the two electron transfers to the cycle have different impacts on the system, and the cycle is more sensitive to the first electron than to the second one. Knowing the dynamics of transient fluctuations, the robustness of the cycle, and the effects from the key interim steps, one has a deeper understanding of the catalytic mechanism of cytochrome P450. PMID- 16706476 TI - Hydration of thermally denatured lysozyme studied by sorption calorimetry and differential scanning calorimetry. AB - We have studied hydration (and dehydration) of thermally denatured hen egg lysozyme using sorption calorimetry. Two different procedures of thermal denaturation of lysozyme were used. In the first procedure the protein was denatured in an aqueous solution at 90 degrees C, in the other procedure a sample that contained 20% of water was denatured at 150 degrees C. The protein denatured at 90 degrees C showed very similar sorption behavior to that of the native protein. The lysozyme samples denatured at 150 degrees C were studied at several temperatures in the range of 25-60 degrees C. In the beginning of sorption, the sorption isotherms of native and denatured lysozyme are almost identical. At higher water contents, however, the denatured lysozyme can absorb a greater amount of water than the native protein due to the larger number of available sorption sites. Desorption experiments did not reveal a pronounced hysteresis in the sorption isotherm of denatured lysozyme (such hysteresis is typical for native lysozyme). Despite the unfolded structure, the denatured lysozyme binds less water than does the native lysozyme in the desorption experiments at water contents up to 34 wt %. Glass transitions in the denatured lysozyme were observed using both differential scanning calorimetry and sorption calorimetry. Partial molar enthalpy of mixing of water in the glassy state is strongly exothermic, which gives rise to a positive temperature dependence of the water activity. The changes of the free energy of the protein induced by the hydration stabilize the denatured form of lysozyme with respect to the native form. PMID- 16706477 TI - A Monte Carlo simulation of the aggregation, phase-separation, and gelation of model globular molecules. AB - Monte Carlo computer simulation on a square 3-D lattice is used to model state behavior of globular copolymers. Two types of globular molecules were defined. One consisted of a single type of subunit (a homopolymer) while the second contained a core of strongly attractive subunits and an outer layer of less strongly attractive subunits (a heteropolymer). Systems of globules were simulated at varied volume fraction (V(F)) and reduced temperature (T(R)), and state diagrams were constructed. These state diagrams contained state boundaries defined by the V(F)/T(R) combinations at which the system formed a percolating network and at which the various component subunits in the globule unfolded. Simulated systems could exist in a number of states (between 4 and 7), depending on the V(F), T(R), whether the molecule was a homo- or heteroglobule and whether the globules were allowed to interact with each other or not. All systems exhibited a gelation/crossover line that resembled a lower critical solution temperature. All systems also exhibited a critical gelation concentration, above which a continuous network was formed. The critical gelation concentration varied between about 2-4% V(F) depending on the type of system. This is comparable to experimental critical gelation concentrations of in the region of 4% (w/w) for a range of associating polymers and biopolymers such as globular proteins and polysaccharides. Other states were formed which included one where elongated, fibril-like aggregated strands were formed, and a micelle-like aggregated state. The results are discussed in terms of the known state behavior of associating polymers and biopolymers (proteins and polysaccharides). PMID- 16706478 TI - Bacteria counting with impedance spectroscopy in a micro probe station. AB - A method to quantify the density of viable biological cells in suspensions is presented. The method is implemented by low-frequency impedance spectroscopy and based on the finding that immobilized ions are released to move freely in the surrounding suspension when viable Escherichia coli cells are killed by a heat shock. The presented results show that an amount of ions corresponding to approximately 2 x 10(8) unit charges are released per viable bacterium killed. A micro probe station with coplanar Ti electrodes was electrically characterized and used as a measuring unit for the impedance spectroscopy recordings. This unit is compatible with common microfabrication techniques and should enable the presented method to be employed using a flow-cell device for viable bacteria counting in miniaturized on-line monitoring systems. PMID- 16706479 TI - Collective behavior of a population of chemically coupled oscillators. AB - Experiments are performed in which a large number (approximately 10(4)) of relaxation oscillators are globally coupled through the concentration of chemicals in the surrounding solution. Each oscillator consists of a microscopic catalyst-loaded particle that displays oscillations in the concentrations of chemical species when suspended in catalyst-free Belousov-Zhabotinsky (BZ) reaction solution. In the absence of stirring, the uncoupled particles display a range of oscillatory frequencies. In the well-stirred system, oscillations appear in the surrounding solution for greater than a critical number density of particles (n(crit)). There is a growth in the amplitude of oscillations with increasing n, accompanied by a slight increase or no change in frequency. A model is proposed to account for the behavior, in which the transfer of activator and inhibitor to and from the bulk medium is considered for each particle. We demonstrate that the appearance and subsequent growth in the amplitude of oscillations may be associated with partial synchronization of the oscillators. PMID- 16706481 TI - Effective screening length of isotropic liquid samples submitted to an applied voltage. AB - A cell of isotropic liquid in the shape of a slab of thickness d and containing ionic impurities is considered. It is shown that the screening effect produced by the ionic charges on the external field is characterized by an effective surface length, lambda(S)(U), depending on the applied voltage U. The analysis indicates that lambda(S)(U)) << lambda(D) when the applied voltage is very large, and lambda(S)(U) --> lambda(D) for very small values of the applied voltage, where lambda(D) is the Debye screening length. The presence of the ions is responsible also for a counterpotential, v, that for small U is such to cancel the effective electric field in the sample, whereas in the opposite limit it is inversely proportional to the applied difference of potential. PMID- 16706480 TI - Charge-induced unilamellar vesicle formation and phase separation in solutions of Di-n-decylmethylamine oxide. AB - A double-tail amine oxide surfactant, di-n-decylmethylamine oxide (2C10MAO), was prepared, and the effects of protonation on aggregate structure were examined by small-angle neutron scattering (SANS), cryo-transmission electron microscopy (cryo-TEM), turbidity, electric conductivity, and solubilization of an oil soluble dye at various degrees of neutralization, X, defined as the mole ratio of HCl/2C10MAO. The surfactant makes an L(2) phase in the nonprotonated state (X = 0) in water. The L(2) phase is in equilibrium with an aqueous L(1) phase. On protonation, unilamellar vesicles (ULVs) are formed over a wide range of compositions (0.05 < X< 0.4-0.5 at C = 10 mM) as observed by cryo-TEM. At X = 0.2, the ULV is stable over a wide concentration range (3 mM < or = C < 0.1 M), but an L(alpha) phase replaces the vesicle phase at C > 0.1 M. SANS results show that the mean radius of the ULV is about 25 nm and the bilayer thickness is about 2 nm, consistent with the extended configuration of the alkyl chains of the surfactant. An important contribution to the enhanced stability of the bilayer structures over the L(2) phase is suggested to be the translational entropy of the counterions. The enhanced stability of the bilayers diminishes as the counterion concentration increases either by an increase of X or by the addition of a salt. When the counterion concentration exceeds a critical value, the ULV solutions transform into the L(2) phase (or L(2)/L(1) two-phase system at low surfactant concentrations). The critical composition X is about 0.4-0.5 in water, but it is below 0.4 in D(2)O. The critical NaCl concentration is below 5 mM at X = 0.2. The stability of ULVs against multilamellar vesicles is ascribed partly to undulation forces and partly to the adjustable nature of the spontaneous curvature of amine oxide monolayers. The characteristics of the ULV of the surfactant remain the same within a temperature range 25-50 degrees C at X = 0.2. An iridescent lamellar phase and possibly an L(3) phase were observed in a very narrow X range (0 < X < 0.02) prior to the vesicle phase. PMID- 16706482 TI - Role of deliquescence lowering in enhancing chemical reactivity in physical mixtures. AB - Mixtures of deliquescent solids are susceptible to deliquescence lowering, where water vapor condensation occurs in mixtures at a lower critical relative humidity (RH(0mix)) than individual component critical relative humidities (RH(0)s). The purpose of this study was to evaluate the effect of deliquescence lowering on chemical reactivity. Sucrose, citric acid and their physical mixtures were characterized using vapor sorption analysis to determine RH(0) and RH(0mix). Acid catalyzed sucrose hydrolysis kinetics was determined using polarimetric analysis. Physical mixtures of sucrose and citric acid crystals were prepared and stored at various relative humidities at 22 degrees C. For these physical mixtures, sucrose hydrolysis was found to occur only when the environmental RH exceeded RH(0mix). Degradation kinetics correlated with the storage RH, being fastest at higher RH. In addition, a lag period was initially observed, which was most prominent for samples stored close to RH(0mix). With exposure to RHs below RH(0mix), no sucrose degradation was detected over the experimental time period. In conclusion, mixtures of deliquescent solids showed increased water sorption at lower RHs, which caused solid dissolution and subsequently led to an increase in the chemical reactivity. PMID- 16706483 TI - Ultrafast dynamics and excited state deactivation of [Ru(bpy)2Sq]+ and its derivatives. AB - Femtosecond transient absorption spectroscopy has been employed to understand the excited state dynamics of [Ru(bpy)(2)Sq](+) (I; bpy is 2,2'-bipyridyl, and Sq is the deprotonated species of the semiquinone form of 1,2-dihydroxy benzene) and its derivatives, a widely studied near-infrared (NIR) active electrochromic dye. Apart from the well-defined dpi(Ru) --> pi(bpy)-based metal-to-ligand charge transfer (MLCT) transition bands at approximately 480 nm, this class of molecules generally shows another dpi(Ru) --> pi(Sq)(SOMO)-based intense MLCT band at around 900 nm, which is known to be redox active and bleaches reversibly upon a change in the oxidation state of the coordinated dioxolene moiety. To have better insight into the photoinduced electron transfer dynamics associated with this MLCT transition, detailed investigations have been carried out on exciting this MLCT band at 800 nm. Immediately after photoexcitation, bleach at 900 nm has been observed, whose recovery is found to follow a triexponential function with major contribution from the ultrafast component. This ultrafast component of approximately 220 fs has been ascribed to the S(1) to S(0) internal conversion process. In addition to the bleach, we have detected two transient species absorbing at 730 and 1000 nm with a formation time approximately 220 fs for both species. The excited state lifetimes for these two transient species have been measured to be 1.5 and 11 ps and have been attributed to excited singlet ((1)MLCT) and triplet ((3)MLCT) states, respectively. Transient measurements carried out on the different but analogous derivatives (II and III) have also shown similar recovery dynamics except that the rate for the internal conversion process has increased with the decrease in the S(1) to S(0) energy gap. The observed results are consistent with the energy gap law for nonradiative decay from S(1) to S(0). PMID- 16706484 TI - Comment on "from subtle to substantial: role of metal ions on pi-pi interactions". PMID- 16706487 TI - Locking down the electronic structure of (monopyrrolo)tetrathiafulvalene in [2]rotaxanes. AB - [reaction: see text] The redox potentials of a highly constrained [2]rotaxane have been measured and used to model the energy of the HOMO of tetrathiafulvalene based bistable [2]rotaxanes in their two co-conformationally isomeric states. Restrained from co-conformational movements, the measured oxidation and reduction potentials provide insights into the orbital energies and electronic structure of a (monopyrrolo)tetrathiafulvalene unit when encircled by a tetracationic cyclobis(paraquat-p-phenylene) ring. PMID- 16706488 TI - Synthesis of the western half of the lolicines and lolitrems. AB - [reaction: see text] The synthesis of the highly substituted indole portion of the complex tremorgenic natural products lolicine A and B is presented. The Diels Alder reaction of a quinone monoimine enables the synthesis of an appropriately substituted indole. The key step in the synthesis is a tandem isopropenyl cuprate addition/aldol cyclocondensation which provides the necessary functionality for elaboration to the 2,2,5,5-tetramethyltetrahydrofuran. PMID- 16706489 TI - Novel synthesis of highly functionalized pyrazolines and pyrazoles by triphenylphosphine-mediated reaction of dialkyl azodicarboxylate with allenic esters. AB - [reaction: see text] Reaction of the Huisgen zwitterion, derived from triphenylphosphine and dialkyl azodicarboxylate, with allenic esters affords highly functionalized pyrazolines and pyrazoles. The crystal structure of pyrazoline derivative 7a showed extensive C-H...O interactions. Pyrazole formation proceeds via a novel nitrogen to carbon migration of the carboalkoxy group. PMID- 16706490 TI - Enantioselective organocatalytic formal [3 + 3]-cycloaddition of alpha,beta unsaturated aldehydes and application to the asymmetric synthesis of (-) isopulegol hydrate and (-)-cubebaol. AB - [reaction: see text] The first highly enantioselective organocatalyzed carbo [3 + 3] cascade cycloaddition of alpha,beta-unsaturated aldehydes is reported. Using this methodology, crotonaldehyde is converted to 6-hydroxy-4-methylcyclohex-1 enecarbaldehyde, which is used in the synthesis of (-)-isopulegol hydrate, (-) cubebaol, and p-tolualdehyde as well as (-)-6-hydroxy-4-methyl-1-cyclohexene-1 methanol acetate, an intermediate in the total synthesis of lycopodium alkaloid magellanine. Other alpha,beta-unsaturated aldehydes give rise to chiral cyclohexadienes via formal [4 + 2] reactions. PMID- 16706491 TI - Discotic liquid crystalline hydrazone compounds: synthesis and mesomorphic properties. AB - [reaction: see text] 1,3,5-Trisacetoacetamidobenzene with three 1,3-diketo groups was synthesized by the reaction of 1,3,5-triaminobenzene with diketene. Discotic hydrazone compounds were prepared by the diazo coupling reaction between 1,3,5 trisacetoacetamidobenzene and diazonium salts of 4-alkyloxyphenylamines. The compounds existed in hydrazone forms exclusively, being stabilized by the intramolecular hydrogen bonds, and showed discotic nematic or columnar hexagonal mesophases. PMID- 16706492 TI - Cyclic tetramer of a metalloporphyrin based on a quadruple hydrogen bond. AB - [reaction: see text] This paper describes the selective formation of a cyclic tetramer from a readily synthesized metalloporphyrin with two self-complementary quadruple hydrogen-bonding units. The extremely strong quadruple hydrogen-bonding unit, 2-ureido-4[1H]-pyrimidinone, enabled the formation of a stable cyclic tetramer based on a tetraphenylporphyrin derivative over a wide concentration range. This hydrogen-bonded tetramer is a new functional unit for use in a higher ordered architecture of a supramolecular porphyrin assembly. PMID- 16706493 TI - Chiral 1,1'-binaphthyl-2,2'-diammonium salt catalysts for the enantioselective Diels-Alder reaction with alpha-acyloxyacroleins. AB - [reaction: see text] A diammonium salt of chiral 1,1'-binaphthyl-2,2'-diamine and trifluoromethanesulfonimide (Tf(2)NH) shows excellent catalytic activity and enantioselectivity for the Diels-Alder reaction of alpha-acyloxyacroleins with cyclic dienes. For example, in the presence of 5 mol % of the ammonium catalyst, the Diels-Alder reaction of alpha-(cyclohexanecarbonyloxy)acrolein with cyclopentadiene proceeded in EtCN at -75 degrees C to give the adducts in 88% yield with 92% exo and 91% ee. This catalyst can be easily prepared in situ by mixing the commercially available chiral diamine and Tf(2)NH. PMID- 16706494 TI - Nonconjugated red-emitting dendrimers with p-type and/or n-type peripheries. AB - [reaction: see text] Dendron-encapsulated materials having a benzo[c][1,2,5]thiadiazole-based red-emitting core and p-type and/or n-type peripheries were synthesized. Intra- and intermolecular energy transfer from the peripheries to the core were found to be peripheries dependent. Red-emitting electroluminescent devices with promising performance were fabricated. PMID- 16706495 TI - Convenient direct synthesis of bisformylated calix[4]arenes via ipso substitution. AB - [reaction: see text] A facile synthesis of bisformylated calix[4]arenes via ipso substitution of p-tert-butylcalix[4]arenes through treatment with hexamethylenetetramine/trifluoroacetic acid is described. Under identical conditions, p-tert-butylcalix[4]arene tetramethyl ether 4 gives proximally substituted bisformylated derivative 4a in a pinched cone conformation. PMID- 16706496 TI - Syntheses, structure, and optical properties of ladder-type fused azaborines. AB - [structure: see text] Ladder-type fused azaborines were synthesized. X-ray crystallographic analysis of a pentacene-type molecule shown here revealed the planarity of a fused azaborine. It was revealed by UV-vis and fluorescence spectra that such fused molecular structures are efficient for the extension of pi-conjugated systems containing main group elements. PMID- 16706497 TI - Efficient C-C double-bond formation reaction via a new synthetic strategy: a self sorting tandem reaction. AB - [reaction: see text] A novel and efficient carbon-carbon double-bond formation reaction via coupling of aryl or heteroaryl methyl ketones has been developed. A dimethyl sulfoxide-iodine-CuO system was proven to be efficient for this reaction and afforded the expected products in good yields. A new synthetic strategy, a self-sorting tandem reaction, was involved in this type of reaction and was presented for the first time. PMID- 16706498 TI - Catalytic enantioselective pyrrole alkylations of alpha,beta-unsaturated 2-acyl imidazoles. AB - [reaction: see text] Enantioselective additions of pyrroles to alpha,beta unsaturated 2-acyl imidazoles catalyzed by the bis(oxazolinyl)pyridine scandium(III) triflate complex (1) have been accomplished. The alpha,beta unsaturated 2-acyl imidazoles were synthesized in high yields through Wittig olefination. A short, enantioselective synthesis of the alkaloid (+)-heliotridane has been accomplished utilizing this methodology and a 2-acyl imidazole cleavage and cyclization. This methodology was then extended to the one-pot asymmetric synthesis of 2-substituted indoles. PMID- 16706499 TI - Synthesis of the aglycone of the shark repellent pavoninin-4 using remote functionalization. AB - [reaction: see text] The aglycone of shark repellent pavoninin-4, (25R)-5alpha cholestan-3alpha,15alpha,26-triol 26-acetate 1a, was synthesized from (25R) cholest-5-en-3beta,26-diol 4 (26-hydroxycholesterol) in eight steps in 18% overall yield. Breslow's remote functionalization strategy was used as a key step to introduce the C-15alpha alcohol on a steroid D ring. An efficient synthesis of the 26-hydroxycholesterol from the 16beta hydroxyl steroid, (25R)-cholest-5-ene 3beta,16beta,26-triol (3a), is also reported. PMID- 16706500 TI - Aerobic intramolecular oxidative amination of alkenes catalyzed by NHC coordinated palladium complexes. AB - [reaction: see text] Palladium(II) complexes bearing a single N-heterocyclic carbene ligand serve as effective catalysts for the aerobic oxidative cyclization of alkenes with pendant sulfonamides. The use of carboxylic acid cocatalysts (AcOH and PhCO(2)H) often leads to significant improvements in catalyst stability and product yield and enables catalytic turnover to be achieved with air, rather than pure oxygen gas, as the source of O(2). PMID- 16706501 TI - New, general, and practical enamine cyclopropanation using dichloromethane. AB - [reaction: see text] Dichloromethane serves as a novel electrophilic carbene equivalent which adds to an enamine double bond. The presence of other alkene moieties in the enamine partner is well tolerated. Even enamines derived from sterically hindered ketones react readily with dichloromethane promoted by TiCl(4)-Mg. PMID- 16706502 TI - Unusual ambiphilic carbenoid equivalent in amide cyclopropanation. AB - [reaction: see text] The titanium-methylene complexes derived from the TiCl(4)-Mg CH(2)Cl(2) system serve as a novel class of ambiphilic carbenoid equivalents, which not only efficiently effect cyclopropanations of a variety amides but also exhibit high chemoselectivity. PMID- 16706503 TI - Novel phloroglucinol derivatives from the roots of Lysidice rhodostegia. AB - [structure: see text] Three novel phloroglucinol derivatives of lysidicins A-C (1 3) have been isolated from the roots of Lysidice rhodostegia and structures were elucidated by comprehensive NMR and MS spectroscopic analysis. 1 and 2 possess spirocyclic benzodihydrofuran skeleton. Their relative stereochemistries were assigned by NOE or NOESY experiment. A plausible pathway for the biosynthesis of 1-3 from 4 and a ketose derivative was postulated. PMID- 16706504 TI - Asymmetric synthesis of polyfunctionalized pyrrolidines from sulfinimine-derived pyrrolidine 2-phosphonates. Synthesis of pyrrolidine 225C. AB - [reaction: see text] The Horner-Wadsworth-Emmons reaction of aldehydes with sulfinimine-derived 3-oxo pyrrolidine phosphonates represents a new method for the asymmetric synthesis of ring-functionalized cis-2,5-disubstituted 3-oxo pyrrolidines. PMID- 16706505 TI - Low ligand loading, highly enantioselective addition of phenylacetylene to aromatic ketones catalyzed by Schiff-base amino alcohols. AB - [reaction: see text] Schiff-base amino alcohols 7a,b derived from L-phenylglycine through three simple steps are found to be highly effective for the enantioselective addition of phenylacetylene to aromatic ketones. When the loading of 7b was 1 mol %, an ee value of up to 95% was obtained. However, when 7b was lowered to 0.1 mol %, a high ee value of 85% was still achieved. A practical solution to synthesize the optically active tertiary propargylic alcohols was described. PMID- 16706506 TI - Nanosized gradient pi-conjugated thienylethynylene dendrimers for light harvesting: synthesis and properties. AB - [reaction: see text] A family of pi-conjugated dendrimers based on truxene and thienylethynylene units are synthesized via a mixed divergent/convergent growth approach. These dendrimers possess an intrinsic energy gradient from the periphery to the core through branches and thus show a broad absorption in the UV vis range and an efficient energy transfer to the lower-energy center. The molecules hence have the potential to be used as light harvesting materials. PMID- 16706507 TI - Flueggenines A and B, two novel C,C-linked dimeric indolizidine alkaloids from Flueggea virosa. AB - [structure: see text] Two unprecedented C,C-linked dimeric indolizidine alkaloids, flueggenines A (1) and B (2), as well as their biosynthetic precursor (-)-norsecurinine, were isolated from the roots of Flueggea virosa. Their structures and absolute configurations were elucidated by spectroscopic methods, especially 2D NMR and CD spectral analyses, and supported by their unique biosynthetic pathway as proposed. Both 1 and 2 were tested against two tumor cell lines, and alkaloid 1 showed weak activity against the P-388 cell line. PMID- 16706508 TI - Enantioselective hydrogenation of acyclic aromatic N-aryl imines catalyzed by an iridium complex of (S,S)-1,2-bis(tert-butylmethylphosphino)ethane. AB - [reaction: see text] An iridium(I) complex of (S,S)-1,2-bis(tert butylmethylphosphino)ethane with tetrakis(3,5-bis(trifluoromethyl)phenyl)borate as the counterion catalyzes the hydrogenation of acyclic aromatic N-aryl imines under 1 atm of hydrogen pressure at room temperature to give the corresponding optically active secondary amines with up to 99% ee. PMID- 16706509 TI - A divergent route to diversity in macromolecules. AB - [reaction: see text] A synthetic route for obtaining functional group diversity in macromolecules is described. The route relies on the differential reactivity of substituted dichlorotriazines. Treatment of a triamine core with substituted dichlorotriazines cleanly yields tris(monochlorotriazines). Subsequent S(N)Ar reactions with amine nucleophiles bearing the functional group of interest yield diversity. If the substituent on the dichlorotriazine is a protected nucleophile, deprotection of the functionalized core allows for iterative reactions and the synthesis of star, dendritic, and hybrid macromolecules. PMID- 16706510 TI - Flavin-based [2]rotaxanes. AB - [reaction: see text] We report the synthesis of flavin-stoppered hydrogen bonded [2]rotaxanes 1 and 2. We also report the electrochemically controllable properties of these systems in solution, and for derivative 2, as an electropolymerized thin film. PMID- 16706511 TI - Semisynthesis and biological evaluation of a novel D-seco docetaxel analogue. AB - [reaction: see text] A 4-methyl-5-oxo docetaxel analogue has been prepared starting from 10-deacetylbaccatin III. This new D-seco docetaxel analogue is slightly less potent than docetaxel at microtubule stabilization in vitro and has about 1/1000th the cytotoxicity of docetaxel. The lack of improved activity for this compound compared to other D-modified taxoids confirms that a C-5 oxygen atom is not required for biological activity. PMID- 16706512 TI - Regio- and stereospecific syntheses and nitric oxide donor properties of (E)-9- and (E)-10-nitrooctadec-9-enoic acids. AB - [reaction: see text] Nitrated fatty acids act as endogenous peroxisome proliferator-activated receptor gamma (PPARgamma) ligands and nitric oxide (NO) donors. We describe the first specific preparation of the two regioisomers of nitrooleic acid, (E)-9-nitrooctadec-9-enoic acid (1) and (E)-10-nitrooctadec-9 enoic acid (2), from cis-cyclooctene and monomethyl azelate, respectively. These syntheses rely upon a Henry condensation between a nine-carbon nitro component and a nine-carbon aldehyde. Preliminary chemiluminescence NO detection studies reveal the ability of these nitrated fatty acids to release NO. PMID- 16706513 TI - Formal total synthesis of the cytotoxic marine ascidian alkaloid haouamine A. AB - [reaction: see text] Described is a convergent 13-step synthesis of a pentacyclic compound which has previously been transformed into haouamine A, therefore constituting a formal total synthesis of this unique marine alkaloid. PMID- 16706514 TI - Synthesis of bowl-shaped polycyclic aromatic hydrocarbons via palladium-catalyzed intramolecular arylation reactions. AB - [reaction: see text] Treatment of the benzannulated enediyne 11 with potassium tert-butoxide in refluxing toluene for 12 h produced 15 via a cascade sequence of cyclization reactions. Two subsequent palladium-catalyzed intramolecular arylation reactions then afforded the bowl-shaped polycyclic aromatic hydrocarbon 16. The X-ray structures of 16 and two closely related molecules show the presence of significant curvatures. PMID- 16706515 TI - Active methylene phosphinic peptides: a new diversification approach. AB - [reaction: see text] Simple, rapid, and efficient methods for P(1)' diversification of phosphinic peptides have been developed, employing alkylation and Knoevenagel-type condensation reactions with active methylene phosphinic scaffolds, thus leading to a wide variety of diversified phosphinic and dehydrophosphinic peptides. PMID- 16706516 TI - Facile synthesis of aryl ether dendrimer from unprotected AB2 building blocks using thionyl chloride as an activating agent. AB - [structure: see text] A novel, rapid, inexpensive, and highly efficient convergent approach for the synthesis of a Frechet- type aryl ether dendrimer using thionyl chloride has been developed. In this method, the purification of each dendron and a dendrimer occurs by recrystallization, extraction, and precipitation. The MALDI-TOF MS spectrum supported the formation of the G2, G3, and G4 dendrons and the star-shaped G4 dendrimer. PMID- 16706517 TI - Aromatic core modified decaphyrins with the largest two-photon absorption cross sections: syntheses and characterization. AB - [structure: see text] Core-modified aromatic decaphyrins with 42pi conjugated electrons exhibit the highest two-photon absorption cross-section value (sigma(2) = 108,000 GM) known for any organic molecule, suggesting possible device applications in the field of nonlinear optics. PMID- 16706518 TI - Judging on host-guest binding mode uniqueness: association entropy as an indicator in enantioselection. AB - [reaction: see text] Supramolecular enantiodifferentiation was studied by isothermal titration calorimetry in an effort to address the order-disorder distinction in the diastereomeric complexes formed from a chiral macrocyclic host and enantiomeric carboxylates. As a result, the association entropy component TDeltaS emerged as an indicator in the enantioselection of tartrate 14 and aspartate 15 by the macrocycle 13 containing two guanidinium anchor groups connected to each other by four urea units. The parent monotopic guanidinium compounds 1 or 2 did not show any enantioselection for chiral carboxylates. PMID- 16706519 TI - Synthesis and properties of an anthraquinone-based redox switch for molecular electronics. AB - [reaction: see text] The synthesis of a molecular wire bearing an anthraquinone core and thioacetyl end groups for gold electrode binding is described. A model anthraquinone system, substituted with tert-butylthio groups, can be reversibly switched electrochemically from cross conjugated (low conductance "off") to linear conjugated (high conductance "on") via two-electron reduction/oxidation reactions. This feature holds promise for the anthraquinone-based wires to be used as redox-controlled switches in molecular electronic devices. PMID- 16706520 TI - Supramolecular chiral recognition by bischlorins: a two-point interaction mode combined with the host's conformational modulation controlled by the guest's stereochemistry and bulkiness. AB - [reaction: see text] Supramolecular chiral recognition based on two-point host guest interactions coupled with the different host's conformational response using the enantiopure bischlorin hosts and different antipodal amine guests is reported. The bulkiness at the guest's stereogenic center controls the chiral recognition properties resulting in switching of the enantioselectivity. PMID- 16706521 TI - Synthesis of a [2]rotaxane incorporating a Ni(II)-salen moiety: evidence of ring opening-and-closing protocol. AB - [reaction: see text] We have synthesized a [2]rotaxane from a crown-ether-like macrocycle that undergoes ring opening and closing through cleavage and formation of imino bonds of a salen moiety; the self-assembly of this macrocycle and a dumbbell-shaped rodlike component, followed by addition of nickel acetate, afforded, after counterion exchange, a [2]rotaxane that is stabilized through coordination of the Ni ion to the macrocycle's salen moiety. PMID- 16706522 TI - Facile preparation of doubly dipyrrolylquinoxaline-bridged expanded porphyrins. Synthesis and structural characterization of an unprecedented [20]tetraphyrin (2.1.2.1). AB - [structure: see text] An unprecedented V-shape (2.1.2.1) expanded porphyrin incorporating an extended pi-conjugated system is described. Its efficient synthesis relied on the use of a new peralkyl tetrapyrrolylquinoxaline building block that constitutes the ideal intermediate for the versatile preparation of new quinoxaline-containing macrocycles. PMID- 16706523 TI - Iron-catalyzed imination of sulfoxides and sulfides. AB - [reaction: see text] The Fe(III)-catalyzed imination of sulfoxides and sulfides with sulfonylamides in the presence of iodinanes has been investigated. The best results were obtained when Fe(acac)(3) was used as a catalyst in combination with iodosylbenzene, providing an effective alternative (stereospecific) access to sulfoximines and sulfilimines. PMID- 16706524 TI - A concise stereoselective synthesis of Preussin, 3-epi-Preussin, and analogues. AB - [reaction: see text] A new stereoselective synthesis of the antifungal and antitumor agents Preussin and 3-epi-Preussin via a Pd-catalyzed carboamination of a protected amino alcohol is described. The key transformation leads to simultaneous formation of the N-C2 bond and the C1'-aryl bond, and allows installation of the aryl group one step from the end of the sequence. This strategy permits the facile construction of a variety of preussin analogues bearing different aromatic groups. PMID- 16706525 TI - Spatial photorelease of oligonucleotides, using a safety-catch photolabile linker. AB - [reaction: see text] We report the development of a safety-catch photolabile linker that allows the light-directed synthesis and spatially selective photorelease of oligonucleotides from microarrays. The linker remains stable to light during DNA synthesis, and is activated for photorelease after acidic hydrolysis. We demonstrate that the photoreleased oligonucleotides can be amplified by PCR to produce double stranded DNA. The advantages offered by this linker could aid the development of an automated gene synthesis platform. PMID- 16706526 TI - Norcystine, a new tool for the study of the structure-activity relationship of peptides. AB - [reaction: see text] Norcysteine (Ncy) is an unnatural amino acid possessing an electronegative sulfur atom directly attached to the alpha-carbon atom. We describe the synthesis of Boc-D,L-Ncy(Mob)-OH, the resolution of its methyl ester, and the introduction of both D- and L-Ncy in GnRH analogues. PMID- 16706527 TI - Synthesis of electron deficient 5,6-aryloxy spiroketals. AB - [reaction: see text] A strategy for the construction of electron deficient 5,6 aryloxy spiroketal is reported. The process should prove useful for the synthesis of natural products containing similar spiroketals. The strategy uncovers an unexpected rearrangement between ortho and para quinone spiroketals. PMID- 16706528 TI - Total synthesis of (-)-mucocin. AB - [reaction: see text] An enantioselective total synthesis of (-)-mucocin has been completed. A combination of asymmetric glycolate aldol additions and ring closing metathesis reactions were exploited to construct the C18-C34 and C7-C17 fragments. A selective cross-metathesis reaction was employed as the key step to couple two complex fragments. PMID- 16706529 TI - General and efficient strategy for erythrinan and homoerythrinan alkaloids: syntheses of (+/-)-3-demethoxyerythratidinone and (+/-)-erysotramidine. AB - [reaction: see text] A general and efficient strategy to both aromatic-type and nonaromatic-type erythrinan and homoerythrinan alkaloids has been developed. This approach involves a key two-step sequence, an alkylation of a ketone with various N-substituted iodoacetamides followed by a N-acyliminium ion promoted intramolecular cyclization, and represents one of the shortest routes to erythrinan and homoerythrinan alkaloids. As the application, the formal total synthesis of (+/-)-3-demethoxyerythratidinone and the total synthesis of (+/-) erysotramidine have been achieved, respectively. PMID- 16706530 TI - Trithiaazafulvalene: a promising building block between tetrathiafulvalene and dithiadiazafulvalene. AB - [reaction: see text] New electroactive building blocks, the trithiaazafulvalenes (TTAFs), were synthesized. These redox-active molecules, intermediate between tetrathiafulvalene (TTF) and dithiadiazafulvalene (DTDAF), show promising features for the elaboration of molecular materials. PMID- 16706531 TI - Synthesis and Ag(I) complexation studies of tethered westiellamide. AB - [reaction: see text] A new tethered macrocyclic ring system based on the natural product westiellamide was prepared to increase the affinity and ease of complexation to Ag(I) ions. NMR and fluorescence Ag(I) titrations confirmed that the tethered macrocycles preserve the unique, high-affinity coordination mode of westiellamide. PMID- 16706532 TI - Synthesis and cofacial pi-stacked packing arrangement of 6,13 bis(alkylthio)pentacene. AB - [reaction: see text] 6,13-Bis(alkylthio)pentacenes directed toward organic field effect transistors (OFETs) were synthesized by the ZnI(2)-mediated reaction of trans-6,13-dihydroxy-6,13-dihydropentacene with alkylthiols, followed by the dehydrogenative aromatization of the resulting trans-6,13-bis(alkylthio)-6,13 dihydropentacenes with p-chloranil. The X-ray crystallographic analysis of 6,13 bis(methylthio)pentacene reveals that this compound is arranged as a result of cofacial pi-stacking with S-S and S-pi interactions. PMID- 16706533 TI - Rapid screening of a receptor with molecular memory. AB - [reaction: see text] Atropisomeric receptor 1 can change conformation and maintain the new conformation when heated and cooled in the presence of a guest molecule. This molecular memory can be used as a rapid method of screening potential guests. Heating atropisomeric diacid 1 with various hydrogen-bonding guests leads to a shift in the syn/anti ratio that could be easily monitored as it is stable at room temperature even in the absence of the guest molecules. PMID- 16706534 TI - Aldolase-catalyzed synthesis of beta-D-galp-(1-->9)-D-KDN: a novel acceptor for sialyltransferases. AB - [reaction: see text] beta-D-Galp-(1-->9)-D-KDN, a disaccharide component of the cell wall of Streptomyces sp. MB-8, was synthesized from beta-D-Galp-(1-->6)-D Manp and pyruvate using a sialic acid aldolase. The obtained KDN-containing compound was a novel acceptor for bacterial sialyltransferases. Unusual alpha2,3- and alpha2,6-linked sialyltrisaccharides and a tetrasaccharide were synthesized using a one-pot two-enzyme system containing a Neisseria meningitidis CMP-sialic acid synthetase and a Pasteurella multocida sialyltransferase or a Photobacterium damsela alpha2,6-sialyltransferase. PMID- 16706535 TI - Highly efficient gold-catalyzed atom-economical annulation of phenols with dienes. AB - [reaction: see text] A highly efficient annulation of phenols and naphthols with dienes was developed by using a combination of AuCl(3)/AgOTf as catalyst. The annulation generated various benzofuran derivatives under mild conditions rapidly. PMID- 16706536 TI - The doubly oxidized, antiaromatic tetraphenylporphyrin complex [Li(TPP)][BF4]. AB - [reaction: see text] Oxidation of dilithium tetraphenylporphyrin, Li(2)(TPP), yields the doubly oxidized porphyrin complex [Li(TPP)][BF(4)]. Free TPP appears to be unstable. The crystal structure of [Li(TPP)][BF(4)] reveals that the 16 atom C-N inner ring of the porphyrin consists of alternating single and double bonds. NMR spectroscopy and nucleus-independent chemical shift (NICS) calculations, in addition to the distortion from planarity observed in the crystal structure, indicate that the 16 pi-electron inner C-N ring of the porphyrin is antiaromatic. PMID- 16706537 TI - Enantioselective one-pot three-component synthesis of propargylamines. AB - [reaction: see text] A copper(I) complex of i-Pr-pybox-diPh has been found to be an efficient catalyst for an enantioselective one-pot three-component synthesis of propargylamines from aldehydes, amines, and alkynes. The reaction has been applied to a wide variety of aromatic aldehydes with excellent yields (up to 99%) and enantiomeric excesses (up to 99% ee). A transition-state model has been proposed to explain the stereochemical outcome of the reaction. PMID- 16706538 TI - Studies on the syntheses of benzoquinone ansamycin antibiotics. Syntheses of the C5-C15 subunits of macbecin I, geldanamycin, and herbimycin A. AB - [reaction: see text] A general and convergent route to the C(5)-C(15) subunits of the benzoquinone ansamycin antibiotics macbecin I, geldanamycin, and herbimycin A is described. Each subunit is prepared by the stepwise coupling of differentially functionalized aldehydes with a pentenyl dianion equivalent derived from diastereoselective pentynylation and regioselective reductive coupling. PMID- 16706539 TI - Rh-catalyzed enantioselective hydrogenation of vinyl boronates for the construction of secondary boronic esters. AB - [reaction: see text] Rh-catalyzed hydrogenation of prochiral vinyl boronates occurs in an enantioselective fashion in the presence of the chiral ligand Walphos 1. This transformation provides access to chiral secondary organoboronates that are not available from alkene hydroboration reactions. The chiral reaction products should be useful in organic synthesis, and preliminary experiments suggest that they may participate in one-pot amination and homologation reactions. PMID- 16706540 TI - Solid-phase synthesis of 1,3-azole-based peptides and peptidomimetics. AB - [reaction: see text] We report highly efficient two-step procedures for the synthesis of 1,3-oxazole-, thiazole-, and imidazole-containing peptides on solid phase from dipeptides composed of C-terminal threonine, serine, cysteine, or diaminopropionic acid by using different cyclodehydration procedures followed or preceded by oxidation. The methods are compatible with Fmoc solid-phase peptide synthesis conditions and with N-Fmoc, N-Boc, N-Cbz, and N-Alloc protecting groups. PMID- 16706541 TI - Verpacamides A-D, a sequence of C11N5 diketopiperazines relating cyclo(Pro-Pro) to cyclo(Pro-Arg), from the marine sponge Axinella vaceleti: possible biogenetic precursors of pyrrole-2-aminoimidazole alkaloids. AB - [reaction: see text] Four C(11)N(5) diketopiperazine metabolites named verpacamides A (6), B (7), C (8), and D (9) consisting of a proline-arginine dipeptide skeleton have been isolated from the marine sponge Axinella vaceleti. Verpacamides A-D are a sequence of metabolites showing the transformation of proline and arginine into the oxidized guanidinyl-cyclo(Pro-Pro) 8 and 9. Compounds 6-9 are structurally and chemically related to C(11)N(5) pyrrole-2 aminoimidazole metabolites also isolated from the Axinellidae and Agelasidae families of sponges and exemplified by dispacamide A (4) and dibromophakellin (10). PMID- 16706542 TI - An efficient direct amination of cyclic amides and cyclic ureas. AB - [reaction: see text] An efficient one-step amination of cyclic amides and ureas has been developed. Treatment of cyclic amides and cyclic ureas with BOP in the presence of DBU in various solvents led to the formation of cyclic amidines and cyclic guanidines in good to excellent yields. Concise syntheses of biologically intriguing kinetin and potent kinase inhibitor olomoucin were thus achieved in just one and two steps, respectively. PMID- 16706543 TI - In pursuit of pestalotiopsin a via zirconocene-mediated ring contraction. AB - [reaction: see text] An asymmetric route from the epimeric beta-hydroxy esters 4 and 5 to the densely functionalized (+)-10 and (-)-10, respectively, is described. Either cyclobutanol can be made available as the predominant product. The levorotatory antipode has been transformed into the advanced intermediate 21 bearing side chains destined to become incorporated into the cyclononene ring of the title compound (1). PMID- 16706544 TI - Catalyst-free conjugated addition of thiols to alpha,beta-unsaturated carbonyl compounds in water. AB - [reaction: see text] Catalyst-free conjugate addition of thiols to alpha,beta unsaturated carbonyl compounds in water is reported. beta-Sulfido carbonyl compounds were formed at room temperature, in short times and with excellent chemoselectivity. Competitive dithiane/dithiolane formation, transesterification, and ester cleavage were not observed. Water played a dual role in simultaneously activating the alpha,beta-unsaturated carbonyl compound and the thiol. This new methodology constitutes an easy, highly efficient, and green synthesis of beta sulfido carbonyl compounds. PMID- 16706545 TI - Highly enantioselective access to primary propargylamines: 4-piperidinone as a convenient protecting group. AB - [reaction: see text] We report a highly enantioselective, catalytic three component coupling of aldehydes, alkynes, and 4-piperidone hydrochloride hydrate to afford the corresponding tertiary propargylamines in useful yields. The selective cleavage of the piperidone protecting group using either ammonia/EtOH or a polymer-supported scavenger amine furnishes primary propargylamines. PMID- 16706546 TI - Allylic alcohol transposition by ortho ester-initiated carbonate extension. synthesis of the vasodilator 11(R),12(S),15(S)-trihydroxyeicosa- 5(Z),8(Z),13(E) trienoic acid. AB - [reaction: see text] The title compound 1 was obtained via methyl ester 2, which was synthesized in four steps from an isomeric 11,14,15-triol ester 5. In the key step, Boc orthoformate 9 was treated with TMS triflate to initiate intramolecular nucleophilic substitution with allylic transposition, forming cyclic carbonates 10 and 11. PMID- 16706548 TI - Shift in the epidemiology of pertussis infection: an indication for pertussis vaccine boosters for adults? AB - Pertussis vaccination of young children has been effective in reducing the overall disease burden due to Bordetella pertussis in many countries. However, the disease has not been eliminated, although humans are the only known host of this pathogen. In fact, in some countries, the number of reported cases has increased dramatically from their nadir and epidemics routinely occur. In areas where >80% of children <2 years of age have been vaccinated, the burden of disease has shifted from elementary school-aged children (who are presumably protected by vaccination) to young infants (<6 months of age) and individuals >11 years of age. With the recent availability of acellular pertussis vaccines for older children to adults, consideration of a change in current vaccination policy is necessary in order to provide better disease control. PMID- 16706549 TI - Nitisinone in the treatment of hereditary tyrosinaemia type 1. AB - Hereditary tyrosinaemia type 1 (HT-1) is a rare genetic disease caused by mutations in the gene for the enzyme fumarylacetoacetase. It usually presents with liver failure but can be manifest as chronic liver disease. Rarely, it may present with nonhepatic manifestations such as renal dysfunction, porphyria-like illness or cardiomyopathy. There is a high lifetime risk of developing hepatocellular carcinoma (HCC). Prior to the development of liver transplantation, most patients died in childhood.The clinical manifestations stem from the cytotoxicity of tyrosine metabolites accumulating proximal to the metabolic defect. Nitisinone acts on tyrosine metabolism upstream of the defect to prevent the production of these metabolites. Nitisinone is used in combination with a tyrosine- and phenylalanine-restricted diet. Nitisinone has transformed the natural history of tyrosinaemia. Liver failure is controlled in 90% of patients, those with chronic liver disease improve and nonhepatic manifestations are abolished. Nitisinone is well tolerated and has few adverse effects other than a predictable rise in plasma tyrosine levels. Nitisinone provides protection against HCC if it is started in infancy, but if commenced after the age of 2 years, a significant risk of HCC remains. Furthermore, where nitisinone is used pre-emptively, liver disease appears to be prevented, suggesting the importance of neonatal screening for tyrosinaemia where possible. Nitisinone is indicated for all children with HT-1, and liver transplantation is only indicated where nitisinone fails, or where the development of HCC is likely or suspected. PMID- 16706550 TI - Gram-positive cocci infections in intensive care: guide to antibacterial selection. AB - The incidence of multiresistance in Gram-positive cocci causing infections in critically ill patients admitted to the intensive care unit (ICU) has increased notably in recent years. Thus, therapeutic proposals have been modified according to the emergence of multiresistant cocci and adapted to epidemiological markers of individual infectious processes, geographical variations of these markers, the availability of new antibacterials, and advances in the knowledge of pharmacokinetic and pharmacodynamic aspects of their use. The current management of critically ill patients should consider new therapeutic approaches, such as the "de-escalating strategy", which includes the administration of empirical antibacterials active against multiresistant pathogens followed by directed treatment based on unequivocal data from antibacterial-susceptibility testing. Optimisation of antibacterial treatment should be viewed in the context of the need to determine plasma drug concentrations, pharmacoeconomic considerations and control of drug-related adverse events. Therapeutic proposals should be developed within the framework of the antibacterial policy of each hospital. The present review is focused on the description of the therapeutic strategies for the main infectious processes caused by Gram-positive cocci in severely ill patients admitted to the ICU according to a review of the pertinent literature and the clinical experience of the authors. PMID- 16706552 TI - Rituximab: a review of its use in non-Hodgkin's lymphoma and chronic lymphocytic leukaemia. AB - Rituximab (MabThera, Rituxan) is an anti-CD20 monoclonal antibody that induces lysis and apoptosis of normal and malignant human B cells, and sensitises malignant B cells to the cytotoxic effect of chemotherapy. In phase III trials in patients with indolent or aggressive B-cell non-Hodgkin's lymphoma (NHL), intravenous rituximab in combination with chemotherapy was more effective as first- or second-line therapy than chemotherapy alone in providing tumour remission and patient survival. Likewise, in patients with chronic lymphocytic leukaemia (CLL), rituximab in combination with chemotherapy appeared more effective than chemotherapy alone as either first- or second-line treatment. In addition, rituximab maintenance therapy was shown to significantly prolong tumour remission and patient survival in patients with indolent B-cell NHL or CLL. The combination of rituximab with cyclophosphamide, doxorubicin, vincristine and prednisone (CHOP) was cost effective as first-line therapy for advanced-stage diffuse large B-cell NHL compared with CHOP alone. Rituximab, either alone or in combination with chemotherapy, was generally well tolerated in patients with NHL or CLL. Overall, rituximab in combination with chemotherapy, is a valuable option for first- and second-line therapy in patients with advanced-stage indolent or aggressive B-cell NHL, and possibly those with B-cell CLL, and is included in current treatment guidelines for these indications. The drug is also potentially useful as maintenance therapy in patients with indolent B-cell NHL or CLL. PMID- 16706551 TI - Substance abuse and psychiatric disorders in HIV-positive patients: epidemiology and impact on antiretroviral therapy. AB - There is a high prevalence of substance abuse and psychiatric disorders among HIV infected individuals. Importantly, drug and alcohol-use disorders are frequently co-morbid with depression, anxiety and severe mental illness. Not only do these disorders increase the risk of contracting HIV, they have also been associated with decreased highly active antiretroviral therapy (HAART) utilisation, adherence and virological suppression. The literature evaluating the relationship between substance abuse and HIV outcomes has primarily focused on injection drug users, although there has been increasing interest in alcohol, cocaine and marijuana. Similarly, the mental health literature has focused largely on depression, with a lesser focus on severe mental illness or anxiety. To date, there is little literature evaluating the association between co-occurring HIV, substance abuse and mental illness on HAART uptake, adherence and virological suppression. Adherence interventions in these populations have demonstrated mixed efficacy. Both directly observed therapy and pharmacist-assisted interventions appear promising, as do integrated behavioural interventions. However, the current intervention literature has several limitations: few of these studies are randomised, controlled trials; the sample sizes have generally been small; and co occurring substance abuse and mental illness has not specifically been targeted in these studies. Future studies examining individual substances of abuse, psychiatric disorders and co-occurring substance abuse and psychiatric disorders on HIV outcomes will inform targeted adherence interventions. PMID- 16706553 TI - Pentosan polysulfate: a review of its use in the relief of bladder pain or discomfort in interstitial cystitis. AB - Pentosan polysulfate (pentosan polysulfate sodium; ELMIRON), a heparin-like, sulfated polysaccharide, is used to manage bladder pain and discomfort in adults with interstitial cystitis (IC). Preliminary clinical models suggest that pentosan polysulfate repairs damaged glycosaminoglycan (GAG) layers lining the urothelium and in vitro data suggest it may provide an anti-inflammatory effect in patients with IC. Pentosan polysulfate shows beneficial effects in a proportion of patients with IC in terms of the improvement of a patient's overall condition and the relief of pain, and it is a generally well tolerated therapy. It is the only US FDA-approved oral treatment for the relief of bladder pain or discomfort associated with IC, and data support its role as an important option in the treatment of patients with IC. PMID- 16706554 TI - Bicalutamide 150mg: a review of its use in the treatment of locally advanced prostate cancer. AB - Bicalutamide (Casodex) is a competitive androgen receptor antagonist that inactivates androgen-regulated prostate cell growth and function, leading to cell apoptosis and inhibition of prostate cancer growth. It is administered orally as a once-daily dose. In the EU and a number of other countries, bicalutamide 150 mg/day is approved in men with locally advanced nonmetastatic prostate cancer as immediate therapy either as an adjuvant to active treatment or as monotherapy as an alternative to surgical or medical castration. Combined analysis of the three trials that comprise the bicalutamide Early Prostate Cancer (EPC) programme showed that bicalutamide administered in conjunction with standard care in men with locally advanced prostate cancer offers disease-free survival benefits over standard care alone and is generally well tolerated. Overall survival was improved to a greater extent in the subgroup of patients who received bicalutamide plus radiation therapy compared with radiation therapy alone. Men with localised prostate cancer do not benefit from the addition of bicalutamide to standard care. Combined analysis of two other studies in men with locally advanced prostate cancer show that bicalutamide monotherapy offers better tolerability and higher health-related quality-of-life (HR-QOL) scores for sexual interest and physical capacity compared with surgical or medical castration, while achieving disease-free and overall survival durations that were not significantly different. Thus, when treatment options are being evaluated, bicalutamide as adjuvant therapy or monotherapy should be considered as an alternative to other available hormonal therapies in men with locally advanced prostate cancer, especially in those who wish to maintain an active lifestyle. PMID- 16706556 TI - Gadofosveset: viewpoints. PMID- 16706555 TI - Gadofosveset. AB - Gadofosveset is the first gadolinium-based magnetic resonance (MR) imaging agent designed to image the blood pool. At clinically relevant concentrations, gadofosveset is highly bound to human serum albumin, which has the effect of increasing its signal-enhancing properties above that of non-protein bound imaging agents, as well as increasing its plasma half-life, allowing for increased imaging times. The use of gadofosveset-enhanced MR angiography was compared with non-enhanced MR angiography in four open-label, multicentre studies in adults with known or suspected arterial disease. Significant improvements in the accuracy and specificity of diagnosis were seen with gadofosveset in patients with aortoiliac disease in two studies. Sensitivity was also improved, with all three readers in one study, and two of three readers in the other study, showing significant improvements with gadofosveset use. Significant improvements in sensitivity, specificity, and accuracy were also seen with gadofosveset in a renal artery disease study. Specificity was significantly improved in patients with pedal artery disease across all readers, with accuracy and sensitivity significantly improved in two of three and one of three readers. Gadofosveset was generally well tolerated in clinical trials, with most adverse events being mild or moderate in severity. PMID- 16706558 TI - Insulin glulisine. AB - Insulin glulisine is a rapid-acting human insulin analogue that has a faster onset of action and shorter duration of action than regular human insulin (RHI) in patients with type 1 or 2 diabetes mellitus and is efficacious in controlling prandial blood glucose levels in these patients. In large, well designed trials in patients with type 1 diabetes, insulin glulisine demonstrated a similar degree of glycaemic control, as measured by glycosylated haemoglobin (HbA(1c)) levels, to RHI after 12 weeks and insulin lispro after 26 weeks. Pre-meal insulin glulisine was also more effective than RHI at controlling 2-hour post-prandial glucose excursions in patients with type 1 or 2 diabetes over a period of 12 weeks. In patients with type 2 diabetes, insulin glulisine induced significantly greater reductions in HbA(1c) levels and 2-hour post-breakfast and post-dinner blood glucose levels than RHI over a period of 26 weeks. Insulin glulisine was generally well tolerated by patients with type 1 or 2 diabetes and had a similar safety profile to insulin lispro or RHI. Severe hypoglycaemia was experienced by similar proportions of insulin glulisine or comparator insulin (insulin lispro or RHI) recipients with type 1 or type 2 diabetes. PMID- 16706559 TI - Insulin glulisine: viewpoints. PMID- 16706562 TI - Lubiprostone. AB - Lubiprostone (Amitiza) is an oral bicyclic fatty acid that selectively activates type 2 chloride channels in the apical membrane of the gastrointestinal epithelium, resulting in increased fluid secretion. In two pivotal, randomised, double-blind, multicentre phase III studies in patients with chronic idiopathic constipation, the frequency of spontaneous bowel movements (SBMs) was significantly greater in patients receiving lubiprostone 24microg twice daily than in those receiving placebo at each weekly timepoint throughout both 4-week studies (p < 0.05). At week 1 in one pivotal trial, the mean frequency of SBMs in the lubiprostone group was 5.9 per week compared with 4.0 per week in the placebo group (p < 0.0001) [baseline SBMs 1.3 and 1.5 per week]. Significantly greater improvements occurred with lubiprostone than placebo in the degree of straining, stool consistency and constipation severity (where reported) in both pivotal studies (p < 0.05 for all comparisons at all timepoints). Lubiprostone was generally well tolerated in clinical trials with no reports of treatment-related serious adverse events in pivotal trials. Nausea was the most common adverse event, occurring in up to 31% of patients receiving lubiprostone. PMID- 16706563 TI - Lubiprostone: viewpoints. PMID- 16706565 TI - Sumatriptan fast-disintegrating/rapid-release tablets. AB - Sumatriptan is a serotonin 5-HT(1B/1D) receptor agonist that is used for the acute treatment of migraine attacks. A new fast-disintegrating/rapid-release sumatriptan tablet (sumatriptan FDT/RRT) has been developed with the goal of speeding absorption and onset of effect compared with standard sumatriptan tablets. Bioequivalence of sumatriptan FDT/RRT tablets to standard sumatriptan tablets was established in healthy volunteers. Initial data suggest that sumatriptan FDT/RRT tablets may disintegrate faster and get emptied from the stomach faster than standard sumatriptan tablets. In a randomised, double-blind, multicentre, early intervention trial in adults with mild migraine, significantly more sumatriptan FDT/RRT 50 and 100mg recipients than placebo recipients were pain free or migraine free 2 hours after receiving study medication. Compared with placebo, pain relief was significantly greater with sumatriptan FDT/RRT 100mg at 25 and 17 minutes following administration, and with sumatriptan FDT/RRT 50mg at 50 and 30 minutes following administration, in two randomised, double blind, multicentre trials in adults with moderate to severe migraine. Sumatriptan FDT/RRT tablets were generally well tolerated; the tolerability profile was similar to that reported for standard sumatriptan tablets in other studies. PMID- 16706566 TI - Sumatriptan fast-disintegrating/rapid-release tablets: viewpoints. PMID- 16706568 TI - Accounting for tastes: a German perspective on the inclusion of patient preferences in healthcare. AB - Internationally, there has been a movement in medicine to better involve patients in decision making, whether it be at the individual or aggregate level. However, the German health sector has been slow to understand and accommodate the preferences of patients. This paper discusses the reasons why and attempts to highlight some of the consequences of not involving patients in healthcare decision making within the German context. We argue that a fundamental paradigm shift at all levels of the German health system, including better doctor/patient communication, is necessary for more choices to be available to patients and to better reflect patient preferences in centralised healthcare decision making. The newly created German health technology assessment agency (IQWiG; Institute for Quality and Economic Efficiency in the Health Care Sector) has indicated that they are interested in focusing on patient preferences, but there is a need for them to clarify what methods they deem suitable for doing so. PMID- 16706570 TI - Cost effectiveness of increasing the dose intensity of chemotherapy with granulocyte colony-stimulating factor in small-cell lung cancer: based on data from the Medical Research Council LU19 trial. AB - BACKGROUND: The use of granulocyte colony-stimulating factor (G-CSF) can enable dose intensification of chemotherapy in small-cell lung cancer (SCLC). However, given its acquisition cost, it is important to assess its cost effectiveness within a resource-constrained health service. OBJECTIVE: To assess the cost effectiveness, from the UK NHS perspective, of G-CSF given in addition to doxorubicin, cyclophosphamide and etoposide (ACE) versus ACE alone in the management of SCLC. METHODS: Using data from a UK Medical Research Council trial (LU19) to assess chemotherapy dose intensification in patients with previously untreated SCLC of any disease extent, a retrospective cost-effectiveness analysis was undertaken. Resource use data, including hospitalisations and non-protocol cancer treatments, were collected during the first 6-month treatment phase of the trial. Mean costs ( pound, 2003 values) of managing patients in the two arms of the trial were calculated. Mean survival duration was calculated for the two groups using patient-specific follow-up data collected in the trial. Incremental cost-effectiveness analysis was undertaken, and uncertainty in cost effectiveness was expressed using cost-effectiveness acceptability curves. RESULTS: The use of G-CSF in addition to ACE chemotherapy is more costly ( 4647 pounds) but results in longer mean survival duration (0.20 years; 0.18 years when discounted). This generates an incremental cost per additional life-year of 25,816 pounds for ACE + G-CSF therapy. The probability of the addition of G-CSF being cost effective, if decision makers are willing to pay 30,000 pounds for an additional life-year, is 0.57. Secondary analysis suggests that cost effectiveness is likely to be sensitive to assumptions about the health-related quality of life (HR-QOL) experienced by patients. CONCLUSION: Based on data collected in the LU19 trial, chemotherapy dose intensification using G-CSF in SCLC adds to health service costs but increases survival duration. Its overall cost effectiveness is likely to be finely balanced. PMID- 16706571 TI - Encouraging smokers to quit: the cost effectiveness of reimbursing the costs of smoking cessation treatment. AB - BACKGROUND: Smoking cessation should be encouraged in order to increase life expectancy and reduce smoking-related healthcare costs. Results of a randomised trial suggested that reimbursing the costs of smoking cessation treatment (SCT) may lead to an increased use of SCT and an increased number of quitters versus no reimbursement. OBJECTIVE: To assess whether reimbursement for SCT is a cost effective intervention (from the Dutch societal perspective), we calculated the incremental costs per quitter and extrapolated this outcome to incremental costs per QALY saved versus no reimbursement. METHODS: In the reimbursement trial, 1266 Dutch smokers were randomly assigned to the intervention or control group using a randomised double consent design. Reimbursement for SCT was offered to the intervention group for a period of 6 months. No reimbursement was offered to the control group. Prolonged abstinence from smoking was determined 6 months after the end of the reimbursement period. The QALYs gained from quitting were calculated until 80 years of age using data from the US. Costs (year 2002 values) were determined from the societal perspective during the reimbursement period (May-November 2002). Benefits were discounted at 4% per annum. The uncertainty of the incremental cost-effectiveness ratios was estimated using non-parametric bootstrapping. RESULTS: Eighteen participants in the control group (2.8%) and 35 participants in the intervention group (5.5%) successfully quit smoking. The costs per participant were 291 euro and 322 euro, respectively. If society is willing to pay 1000 euro or 10,000 euro for an additional 12-month quitter, the probability that reimbursement for SCT would be cost effective was 50% or 95%, respectively. If society is willing to pay 18,000 euro for a QALY, the probability that reimbursement for SCT would be cost effective was 95%. However, the external validity of the extrapolation from quitters to QALYs is uncertain and several assumptions had to be made. CONCLUSION: Reimbursement for SCT may be cost effective if Dutch society is willing to pay 10,000 euro for an additional quitter or 18,000 euro for a QALY. PMID- 16706569 TI - Cost effectiveness of preventive interventions in type 2 diabetes mellitus: a systematic literature review. AB - A systematic review of the literature was conducted to give an overview of economic evaluations of preventive interventions in type 2 diabetes mellitus. The interventions were sorted by type of preventive intervention (primary, secondary or tertiary) and by category (e.g. education, medication for hypertension). Several databases were searched for studies published between January 1990 and May 2004 on the three types of preventive intervention. For each study selected, inclusion of specific components from a standardised list of items, including quality, was recorded in a database. Summary tables were generated based on the database.A number of conclusions were drawn from this review. The most important was that strict blood pressure control was a more cost-effective intervention than less strict control, as shown by six studies reporting cost savings to very low costs per life-year gained. Primary and secondary prevention of type 2 diabetes were also highly cost effective, but these results were based on very few studies. Medications to reduce weight and hyperglycaemia together were cost effective compared with conventional interventions. Finally, the separate results regarding medications to reduce weight, hyperglycaemia and hypercholesterolaemia varied enormously, thus no conclusion could be drawn and further economic analysis is required. PMID- 16706572 TI - Comprehensive decision-analytic model and Bayesian value-of-information analysis: pentoxifylline in the treatment of chronic venous leg ulcers. AB - OBJECTIVE: To conduct a Bayesian value-of-information analysis of the cost effectiveness of pentoxifylline (vs placebo) as an adjunct to compression for venous leg ulcers. METHODS: A probabilistic Markov model was developed to estimate mean clinical benefits and costs associated with oral pentoxifylline (400mg three times daily) and placebo. Clinical data were obtained from a systematic review and synthesised using Bayesian methods. The decision uncertainty associated with the adoption of pentoxifylline as well as the maximum value associated with further research were estimated before and after the completion of the largest 'definitive' treatment trial. Resource use was obtained from a UK national audit and unit costs applied (pounds, 2004 values). RESULTS: The prior and posterior analyses suggest that pentoxifylline is a dominant therapy versus placebo. In the prior analysis, patients in the pentoxifylline group healed an average of 8.28 weeks quicker than patients in the placebo group (95% credibility interval [CI] 1.89, 14.56), had a 0.02 gain in QALYs (95% CI 0.12, 0.17) and an average reduction in cost of 153.4 pounds (95% CI -53.11, 354.9). Estimates of the uncertainty surrounding the cost effectiveness of pentoxifylline and the value of perfect information in both analyses did not suggest further research was justified. In the prior analysis, for willingness-to pay values of 0 pounds, 100 pounds and 500 pounds per QALY gained, the estimated values of perfect information were 128,200 pounds, 127,100 pounds and 126,700 pounds, respectively. Incorporation of the information from the largest randomised controlled trial on pentoxifylline did improve the estimate of the clinical effect associated with this drug; however, the variation was not large enough to reverse either the decision regarding the dominance of pentoxifylline or the maximum value associated with further research. CONCLUSION: Bayesian value of-information analysis represents a valuable tool for healthcare decision making. Had the results from this analysis been available before the largest trial was funded, a more efficient allocation of research and development resources could have been made. PMID- 16706573 TI - Economic evaluation of weekly epoetin alfa versus biweekly darbepoetin alfa for chemotherapy-induced anaemia: evidence from a 16-week randomised trial. AB - INTRODUCTION: A 16-week, open-label, multicentre, randomised trial of weekly epoetin alfa 40 000 units versus biweekly darbepoetin alfa 200microg among 358 patients with solid-tumour cancers and chemotherapy-induced anaemia demonstrated superior haematological outcomes with epoetin alfa. We sought to compare resource use, costs and clinical outcomes between treatment groups and report the results using a cost-consequences framework. METHODS: Pre-specified methods were used to assign costs (US dollars, year 2004-5 values) to medical resources and patient time using a societal perspective. Costs for inpatient care, outpatient care and physician services were based on US Medicare reimbursement rates. Indirect costs assigned to patient time spent receiving study medication were based on the mean hourly wage in the US. In the base-case analysis, the average wholesale price was used to assign costs to medications. Clinical outcomes included all haemoglobin levels and transfusions recorded throughout the trial. Sensitivity analyses were performed to evaluate the impact of different costing methods, cost sources, perspectives and methods to assign haemoglobin values following a blood transfusion. RESULTS: Over a mean follow-up duration of 11.8 weeks, the average cost of study medications and their administration was the single largest component of total costs and was similar between groups (epoetin alfa 5979 US dollars and darbepoetin alfa 5935 US dollars, difference 44 US dollars; 95% CI 590, 692). There were no significant differences in the proportions of patients hospitalised (epoetin alfa 24.6%, darbepoetin alfa 22.0%; p = 0.57). Patients randomised to epoetin alfa experienced more inpatient days, on average, than patients randomised to darbepoetin alfa (2.6 vs 1.6, 95% CI for the difference, 0.07, 2.27). However, with regard to transfusions, patients in the epoetin alfa arm required fewer units of blood than patients in the darbepoetin alfa arm (0.46 vs 0.88, 95% CI for the difference -0.77, -0.08). Mean total costs, comprising costs for study medications and their administration, inpatient care, transfusions, unplanned radiation therapy, haematology and laboratory services, chemotherapy and non-chemotherapy drugs and indirect costs were 14,976 US dollars in the epoetin alfa arm compared with 14,101 US dollars in the darbepoetin alfa arm, a difference of 875 US dollars (95% CI for difference -849, 2607), of which 98% of the difference was attributable to higher inpatient costs in the epoetin alfa arm (2374 US dollars vs 1520 US dollars; 95% CI for difference -33, 1955). Assessments of multiple clinical measures demonstrated improved outcomes with epoetin alfa relative to darbepoetin alfa. CONCLUSION: Most clinical outcome measures suggested greater improvement with epoetin alfa relative to darbepoetin alfa, but most costs for both agents appeared similar. Decision makers must evaluate the differences in costs and efficacy measures that are most relevant from their perspectives. PMID- 16706575 TI - Cost considerations in the medical management of glaucoma in the US: estimated yearly costs and cost effectiveness of bimatoprost compared with other medications. PMID- 16706574 TI - Letrozole: a pharmacoeconomic review of its use in postmenopausal women with breast cancer. AB - Letrozole (Femara), an aromatase inhibitor that blocks estrogen synthesis by inhibiting the final step of the estrogen biosynthetic pathway, is approved for use in a wide range of breast cancer settings. Randomised clinical trials in postmenopausal women with hormone-responsive early-stage breast cancer have demonstrated that, as adjuvant therapy, letrozole has greater efficacy than tamoxifen. It is also more effective than placebo as extended adjuvant therapy after completion of tamoxifen therapy in these patients. In women with hormone responsive advanced breast cancer, letrozole is superior to tamoxifen in prolonging the time to disease progression and time to treatment failure in a first-line setting, and is at least as effective as anastrozole and more effective than megestrol for some endpoints (in one of two trials) in a second line setting. Letrozole is generally well tolerated, and in a health-related quality-of-life analysis from a large clinical trial, patient well-being with letrozole as extended adjuvant therapy did not differ from that with placebo. Modelled analyses from the UK and the US suggest that, in postmenopausal women with hormone-receptor-positive early-stage breast cancer, letrozole is likely to be a cost-effective alternative to tamoxifen as adjuvant therapy; moreover, using letrozole as extended adjuvant therapy after tamoxifen, rather than no further treatment, is also a cost-effective treatment strategy. Sensitivity analyses have shown these results to be robust. In terms of direct healthcare costs, pharmacoeconomic models suggest that letrozole is a cost-effective alternative to tamoxifen as first-line therapy in postmenopausal women with hormone-responsive advanced breast cancer from the perspectives of the UK NHS, the Canadian and Italian public healthcare systems and the Japanese national health insurance system. Incremental costs per QALY or progression-free year gained over tamoxifen were well within the recommended limits for acceptability of new agents that are more effective and more expensive than existing therapies in the UK, Japan and Canada. Modelled analyses from the UK and Canada have also suggested that letrozole is cost effective as second-line therapy for advanced breast cancer in postmenopausal women who have disease progression following anti-estrogen therapy. In conclusion, letrozole is an effective and well tolerated treatment for postmenopausal women with early-stage or advanced hormone-responsive breast cancer. Pharmacoeconomic analyses from UK and North American perspectives support the use of letrozole in hormone-responsive early-stage breast cancer in both the adjuvant and extended adjuvant settings. In addition, other modelled analyses conducted in a variety of healthcare systems across different countries consistently suggest that letrozole is cost effective in advanced treatment settings. PMID- 16706577 TI - Application of fracture mechanics to failure in manatee rib bone. AB - BACKGROUND: The Florida manatee (Trichechus manatus latirostris) is listed as endangered by the U.S. Department of the Interior. Manatee ribs have different microstructure from the compact bone of other mammals. Biomechanical properties of the manatee ribs need to be better understood. Fracture toughness (K(C)) has been shown to be a good index to assess the mechanical performance of bone. Quantitative fractography can be used in concert with fracture mechanics equations to identify fracture initiating defects/cracks and to calculate the fracture toughness of bone materials. METHOD OF APPROACH: Fractography is a standard technique for analyzing fracture behavior of brittle and quasi-brittle materials. Manatee ribs are highly mineralized and fracture in a manner similar to quasi-brittle materials. Therefore, quantitative fractography was applied to determine the fracture toughness of manatee ribs. RESULTS: Average fracture toughness values of small flexure specimens from six different sizes of manatees ranged from 1.3 to 2.6 MPa(m)(12). Scanning electron microscope (SEM) images show most of the fracture origins were at openings for blood vessels and interlayer spaces. CONCLUSIONS: Quantitative fractography and fracture mechanics can be combined to estimate the fracture toughness of the material in manatee rib bone. Fracture toughness of subadult and calf manatees appears to increase as the size of the manatee increases. Average fracture toughness of the manatee rib bone materials is less than the transverse fracture toughness of human and bovine tibia and femur. PMID- 16706579 TI - Vibrational characteristics of bone fracture and fracture repair: application to excised rat femur. AB - BACKGROUND: The vibrational characteristics of any object are directly dependent on the physical properties of that object. Therefore, changing the physical properties of an object will cause the object to adopt changed natural frequencies. A fracture in a bone results in the loss of mechanical stability of the bone. This change in mechanical properties of a bone should result in a change of the resonant frequencies of that bone. A vibrational method for bone evaluation has been introduced. METHOD OF APPROACH: This method uses the radiation force of focused amplitude-modulated ultrasound to exert a vibrating force directly, and remotely, on a bone. The vibration frequency is varied in the range of interest to induce resonances in the bone. The resulting bone motion is recorded and the resonance frequencies are determined. Experiments are conducted on excised rat femurs and resonance frequencies of intact, fractured, and bonded (simulating healed) bones are measured. RESULTS: The experiments demonstrate that changes in the resonance frequency are indicative of bone fracture and healing, i.e., the fractured bone exhibits a lower resonance frequency than the intact bone, and the resonance frequency of the bonded bone approaches that of the intact bone. CONCLUSION: It is concluded that the proposed radiation force method may be used as a remote and noninvasive tool for monitoring bone fracture and healing process, and the use of focused ultrasound enables one to selectively evaluate individual bones. PMID- 16706578 TI - A 3D computational simulation of fracture callus formation: influence of the stiffness of the external fixator. AB - The stiffness of the external fixation highly influences the fracture healing pattern. In this work we study this aspect by means of a finite element model of a simple transverse mid-diaphyseal fracture of an ovine metatarsus fixed with a bilateral external fixator. In order to simulate the regenerative process, a previously developed mechanobiological model of bone fracture healing was implemented in three dimensions. This model is able to simulate tissue differentiation, bone regeneration, and callus growth. A physiological load of 500 N was applied and three different stiffnesses of the external fixator were simulated (2300, 1725, and 1150 N/mm). The interfragmentary strain and load sharing mechanism between bone and the external fixator were compared to those recorded in previous experimental works. The effects of the stiffness on the callus shape and tissue distributions in the fracture site were also analyzed. We predicted that a lower stiffness of the fixator delays fracture healing and causes a larger callus, in correspondence to well-documented clinical observations. PMID- 16706580 TI - Prediction of cortical bone elastic constants by a two-level micromechanical model using a generalized self-consistent method. AB - A two-level micromechanical model of cortical bone based on a generalized self consistent method was developed to take into consideration the transversely isotropic elasticity of many microstructural features in cortical bone, including Haversian canals, resorption cavities, and osteonal and interstitial lamellae. In the first level, a single osteon was modeled as a two-phase composite such that Haversian canals were represented by elongated pores while the surrounding osteonal lamellae were considered as matrix. In the second level, osteons and resorption cavities were modeled as multiple inclusions while interstitial lamellae were regarded as matrix. The predictions of cortical bone elasticity from this two-level micromechanical model were mostly in agreement with experimental data for the dependence of transversely isotropic elasticity of human femoral cortical bone on porosity. However, variation in cortical bone elastic constants was greater in experimental data than in model predictions. This could be attributed to variations in the elastic properties of microstructural features in cortical bone. The present micromechanical model of cortical bone will be useful in understanding the contribution of cortical bone porosity to femoral neck fractures. PMID- 16706581 TI - Structural mechanisms in the abolishment of VEGF-induced microvascular hyperpermeability by cAMP. AB - To investigate the structural mechanisms by which elevation of the intraendothelial cAMP levels abolishes or attenuates the transient increase in microvascular permeability by vascular endothelial growth factor (VEGF), we examined cAMP effect on VEGF-induced hyperpermeability to small solute sodium fluorescein (Stokes radius = 0.45 nm) P(sodium fluorescein), intermediate-sized solute alpha-lactalbumin (Stokes radius = 2.01 nm) P(alpha-lactalbumin), and large solute albumin (BSA, Stokes radius = 3.5 nm) P(BSA) on individually perfused microvessels of frog mesenteries. After 20 min pretreatment of 2 mM cAMP analog, 8-bromo-cAMP, the initial increase by 1 nM VEGF was completely abolished in P(sodium fluorescein) (from a peak increase of 2.6+/-0.37 times control with VEGF alone to 0.96+/-0.07 times control with VEGF and cAMP), in P(alpha lactalbumin) (from a peak increase of 2.7+/-0.33 times control with VEGF alone to 0.76+/-0.07 times control with VEGF and cAMP), and in P(BSA) (from a peak increase of 6.5+/-1.0 times control with VEGF alone to 0.97+/-0.08 times control with VEGF and cAMP). Based on these measured data, the prediction from our mathematical models suggested that the increase in the number of tight junction strands in the cleft between endothelial cells forming the microvessel wall is one of the mechanisms for the abolishment of VEGF-induced hyperpermeability by cAMP. PMID- 16706582 TI - Large Negative Stress Phase Angle (SPA) attenuates nitric oxide production in bovine aortic endothelial cells. AB - Hemodynamics plays an important role in cardiovascular physiology and pathology. Pulsatile flow (Q), pressure (P), and diameter (D) waveforms exert wall shear stress (WSS), normal stress, and circumferential strain (CS) on blood vessels. Most in vitro studies to date have focused on either WSS or CS but not their interaction. Recently, we have shown that concomitant WSS and CS affect EC biochemical response modulated by the temporal phase angle between WSS and CS (stress phase angle, SPA). Large negative SPA has been shown to occur in regions of the circulation where atherosclerosis and intimal hyperplasia are prevalent. Here, we report that nitric oxide (NO) biochemical secretion was significantly decreased in response to a large negative SPA of -180 deg with respect to an SPA of 0 degrees in bovine aortic endothelial cells (BAEC) at 5 h. A new hemodynamic simulator for the study of the physiologic SPA was used to provide the hemodynamic conditions of pro-atherogenic (SPA = -180 deg) and normopathic (SPA = 0 deg) states. The role of complex hemodynamics in vascular remodeling, homeostasis, and pathogenesis can be advanced by further assessment of the hypothesis that a large negative SPA is pro-atherogenic. PMID- 16706583 TI - Forced and natural convective drying of trehalose/water thin films: implication in the desiccation preservation of Mammalian cells. AB - Trehalose is believed to offer desiccation protection to mammalian cells by forming stable glassy matrices. The goal of the current study was to explore the desiccation kinetics of thin films of trehalose-water solution under forced and natural convective conditions and to investigate the thermophysical state of mammalian cells at the bottom of the thin film. We developed a finite difference model based on the mass and energy conservation equations coupled to the water transport model from the cells. The boundary conditions were obtained from correlations or experimental measurements and the Gordon-Taylor equation was used to predict the glass transition temperature at every location. Results indicated that there are three distinct regimes for drying for both forced and natural convection, characterized by the slope of the moisture content plot as a function of time. Our results also indicate that the surface of the solution reached the glassy state in less than 10 min for the Reynolds (forced) numbers explored and approximately 30 min for some Rayleigh (natural convective) numbers; however, significant water was trapped at this instant. Larger drying force hastened quicker glass formation but trapped more water. The numerical model was capable of predicting the drying kinetics for the dilute region accurately, but deviated while predicting the other regimes. Based on these experimental validations of the model, the osmotic response of different cells located at the bottom of the solution with orders of magnitude difference in their membrane permeability (Lp) was predicted. The results suggested that extracellular glass formed around cells at the bottom of a trehalose-water solution by the propagation of glass into the solution; however it takes more than an order of magnitude time (approximately 7 min to >100 min for forced convective drying) to remove sufficient water to form glass around cells from the time when the first surface glass is formed. This is attributed to low diffusivity of water through the glass. In addition, the water transport from the glassy matrix could be either diffusion or Lp limited. For diffusion-limited transport, lowering the film thickness at the beginning of drying by half almost lowers the drying time by an order of magnitude. In summary, the optimal design of convective desiccation protocols requires accounting for the size of the cell, their membrane permeability (Lp) and the starting thickness of the solution. PMID- 16706584 TI - Developing pulsatile flow in a deployed coronary stent. AB - A major consequence of stent implantation is restenosis that occurs due to neointimal formation. This patho-physiologic process of tissue growth may not be completely eliminated. Recent evidence suggests that there are several factors such as geometry and size of vessel, and stent design that alter hemodynamic parameters, including local wall shear stress distributions, all of which influence the restenosis process. The present three-dimensional analysis of developing pulsatile flow in a deployed coronary stent quantifies hemodynamic parameters and illustrates the changes in local wall shear stress distributions and their impact on restenosis. The present model evaluates the effect of entrance flow, where the stent is placed at the entrance region of a branched coronary artery. Stent geometry showed a complex three-dimensional variation of wall shear stress distributions within the stented region. Higher order of magnitude of wall shear stress of 530 dyn/cm2 is observed on the surface of cross link intersections at the entrance of the stent. A low positive wall shear stress of 10 dyn/cm2 and a negative wall shear stress of -10 dyn/cm2 are seen at the immediate upstream and downstream regions of strut intersections, respectively. Modified oscillatory shear index is calculated which showed persistent recirculation at the downstream region of each strut intersection. The portions of the vessel where there is low and negative wall shear stress may represent locations of thrombus formation and platelet accumulation. The present results indicate that the immediate downstream regions of strut intersections are areas highly susceptible to restenosis, whereas a high shear stress at the strut intersection may cause platelet activation and free emboli formation. PMID- 16706585 TI - Numerical analysis of the hemodynamics and embolus capture of a greenfield vena cava filter. AB - BACKGROUND: Vena Cava filters are used to prevent pulmonary embolism in patients with deep vein thrombosis who are unresponsive to anticoagulation therapy. Various filter designs exist in the market with different characteristics distinguishing them. An understanding of the characteristics of these filters is desirable in order to develop better designs. METHODS: A computational fluid dynamical study of the flow over an unoccluded stainless steel Greenfield Vena Cava filter (Boston Scientific, Watertown, MA) to determine its properties has been performed. Simulation of flow over a filter placed axisymmetrically in a rounded inferior vena cava has been performed at a Reynolds numbers of 1000 and the consequences of the flow (by studying parameters like shear stress and stagnation zones) have been discussed. Furthermore, a new finite element based numerical method has been developed that allows the study of capturing properties of Inferior Vena Cava filters. The key idea is the introduction of a thin-wire model (TWM) that enables a drastic reduction in the computational cost while still maintaining control on the physics of the problem. This numerical technique has been applied to evaluate the embolus capture characteristic of a Greenfield filter. RESULTS: The flow around the unoccluded filter is found to be steady and laminar at the conditions studied. A recirculation/stagnation zone develops immediately downstream of the filter head. This zone is significantly larger when the central hole is occluded. The shear stress and stagnation zone properties for such a flow over a Greenfield filter are compared with existing literature (in vitro studies). A graph showing the regions wherein clots escape or get captured has been determined by a means of numerical simulations. The data has further been analyzed to determine the probability of clot capture as function of the clot size. CONCLUSIONS: The stagnation zone formed behind the head of the Greenfield filter is found to be smaller in size when compared to that of the same filter with the central hole occluded. A map of the shear stress distribution shows a small region having the potential for thrombogenesis. The non-Newtonian properties of blood are not seen to cause much variation in the flow field when compared to the Newtonian model. However variation in the cava size leads to a significant change in the shear stresses. This study also establishes a novel method wherein computational means are used to determine the efficacy of clot capturing of filters. These techniques can further be used to compare the different characteristics among filters. PMID- 16706586 TI - Inlet conditions for image-based CFD models of the carotid bifurcation: is it reasonable to assume fully developed flow? AB - BACKGROUND: Computational fluid dynamics tools are useful for their ability to model patient specific data relevant to the genesis and progression of atherosclerosis, but unavailable to measurement tools. The sensitivity of the physiologically relevant parameters of wall shear stress (WSS) and the oscillatory shear index (OSI) to secondary flow in the inlet velocity profiles was investigated in three realistic models of the carotid bifurcation. METHOD OF APPROACH: Secondary flow profiles were generated using sufficiently long entrance lengths, to which curvature and helical pitch were added. The differences observed were contextualized with respect to effect of the uncertainty of the models' geometry on the same parameters. RESULTS: The effects of secondary velocities in the inlet profile on WSS and OSI break down within a few diameters of the inlet. Overall, the effect of secondary inlet flow on these models was on average more than 3.5 times smaller than the effect of geometric variability, with 13% and 48% WSS variability induced by inlet secondary flow and geometric differences, respectively. CONCLUSIONS: The degree of variation is demonstrated to be within the range of the other computational assumptions, and we conclude that given a sufficient entrance length of realistic geometry, simplification to fully developed axial (i.e., Womersley) flow may be made without penalty. Thus, given a choice between measuring three components of inlet velocity or a greater geometric extent, we recommend effort be given to more accurate and detailed geometric reconstructions, as being of primary influence on physiologically significant indicators. PMID- 16706587 TI - Flow visualization and acoustic consequences of the air moving through a static model of the human larynx. AB - Flow visualization with smoke particles illuminated by a laser sheet was used to obtain a qualitative description of the air flow structures through a dynamically similar 7.5x symmetric static scale model of the human larynx (divergence angle of 10 deg, minimal diameter of 0.04 cm real life). The acoustic level downstream of the vocal folds was measured by using a condenser microphone. False vocal folds (FVFs) were included. In general, the glottal flow was laminar and bistable. The glottal jet curvature increased with flow rate and decreased with the presence of the FVFs. The glottal exit flow for the lowest flow rate showed a curved jet which remained laminar for all geometries. For the higher flow rates, the jet flow patterns exiting the glottis showed a laminar jet core, transitioning to vortical structures, and leading spatially to turbulent dissipation. This structure was shortened and tightened with an increase in flow rate. The narrow FVF gap lengthened the flow structure and reduced jet curvature via acceleration of the flow. These results suggest that laryngeal flow resistance and the complex jet flow structure exiting the glottis are highly affected by flow rate and the presence of the false vocal folds. Acoustic consequences are discussed in terms of the quadrupole- and dipole-type sound sources due to ordered flow structures. PMID- 16706588 TI - How changing the inversion/eversion foot angle affects the nondriving intersegmental knee moments and the relative activation of the vastii muscles in cycling. AB - Nondriving intersegmental knee moment components (i.e., varus/valgus and internal/external axial moments) are thought to be primarily responsible for the etiology of overuse knee injuries such as patellofermoral pain syndrome in cycling because of their relationship to muscular imbalances. However the relationship between these moments and muscle activity has not been studied. Thus the four primary objectives of this study were to test whether manipulating the inversion/eversion foot angle alters the varus/valgus knee moment (Objective 1) and axial knee moment (Objective 2) and to determine whether activation patterns of the vastus medialis oblique (VMO), vastus lateralis (VL), and tensor fascia latae (TFL) were affected by changes in the varus/valgus (Objective 3) and axial knee moments (Objective 4). To fulfill these objectives, pedal loads and lower limb kinematic data were collected from 15 subjects who pedaled with five randomly assigned inversion/eversion angles: 10 deg and 5 deg everted and inverted and 0 deg (neutral). A previously described mathematical model was used to compute the nondriving intersegmental knee moments throughout the crank cycle. The excitations of the VMO, VL, and TFL muscles were measured with surface electromyography and the muscle activations were computed. On average, the 10-deg everted position decreased the peak varus moment by 55% and decreased the peak internal axial moment by 53% during the power stroke (crank cycle region where the knee moment is extensor). A correlation analysis revealed that the VMO/VL activation ratio increased significantly and the TFL activation decreased significantly as the varus moment decreased. For both the VMO/VL activation ratio and the TFL activation, a path analysis indicated that the varus/valgus moment was highly correlated to the axial moment but that the correlation between muscle activation and the varus moment was due primarily to the varus/valgus knee moment rather than the axial knee moment. The conclusion from these results is that everting the foot may be beneficial towards either preventing or ameliorating patellofemoral pain syndrome in cycling. PMID- 16706589 TI - Sensitivities of medial meniscal motion and deformation to material properties of articular cartilage, meniscus and meniscal attachments using design of experiments methods. AB - This study investigated the role of the material properties assumed for articular cartilage, meniscus and meniscal attachments on the fit of a finite element model (FEM) to experimental data for meniscal motion and deformation due to an anterior tibial loading of 45 N in the anterior cruciate ligament-deficient knee. Taguchi style L18 orthogonal arrays were used to identify the most significant factors for further examination. A central composite design was then employed to develop a mathematical model for predicting the fit of the FEM to the experimental data as a function of the material properties and to identify the material property selections that optimize the fit. The cartilage was modeled as isotropic elastic material, the meniscus was modeled as transversely isotropic elastic material, and meniscal horn and the peripheral attachments were modeled as noncompressive and nonlinear in tension spring elements. The ability of the FEM to reproduce the experimentally measured meniscal motion and deformation was most strongly dependent on the initial strain of the meniscal horn attachments (epsilon(1H)), the linear modulus of the meniscal peripheral attachments (E(P)) and the ratio of meniscal moduli in the circumferential and transverse directions (E(theta)E(R)). Our study also successfully identified values for these critical material properties (epsilon(1H) = -5%, E(P) = 5.6 MPa, E(theta)E(R) = 20) to minimize the error in the FEM analysis of experimental results. This study illustrates the most important material properties for future experimental studies, and suggests that modeling work of meniscus, while retaining transverse isotropy, should also focus on the potential influence of nonlinear properties and inhomogeneity. PMID- 16706590 TI - Directional, regional, and layer variations of mechanical properties of esophageal tissue and its interpretation using a structure-based constitutive model. AB - The esophagus, like other soft tissues, exhibits nonlinear and anisotropic mechanical properties. As a composite structure, the properties of the outer muscle and inner mucosal layer are different. It is expected that the complex mechanical properties will induce nonhomogeneous stress distributions in the wall and nonuniform tissue remodeling. Both are important factors which influence the function of mechanosensitive receptor located in various layers of the wall. Hence, the characterization of the mechanical properties is essential to understand the neuromuscular motion of the esophagus. In this study, the uniaxial tensile tests were conducted along two mutually orthogonal directions of porcine esophageal tissue to identify the directional (circumferential and axial), regional (abdominal, thoracic, and cervical), and layer (muscle and mucosa) variations of the mechanical properties. A structure-based constitutive model, which took the architectures of the tissue's microstructures into account, was applied to describe the mechanical behavior of the esophagus. Results showed that the constitutive model successfully described the mechanical behavior and provided robust estimates of the material parameters. In conclusion, the model was demonstrated to be a good descriptor of the mechanical properties of the esophagus and it was able to facilitate the directional, layer, and regional comparisons of the mechanical properties in terms of the associated material parameters. PMID- 16706591 TI - Microplane constitutive model and computational framework for blood vessel tissue. AB - This paper presents a nonlinearly elastic anisotropic microplane formulation in 3D for computational constitutive modeling of arterial soft tissue in the passive regime. The constitutive modeling of arterial (and other biological) soft tissue is crucial for accurate finite element calculations, which in turn are essential for design of implants, surgical procedures, bioartificial tissue, as well as determination of effect of progressive diseases on tissues and implants. The model presented is defined at a lower scale (mesoscale) than the conventional macroscale and it incorporates the effect of all the (collagen) fibers which are anisotropic structural components distributed in all directions within the tissue material in addition to that of isotropic bulk tissue. It is shown that the proposed model not only reproduces Holzapfel's recent model but also improves on it by accounting for the actual three-dimensional distribution of fiber orientation in the arterial wall, which endows the model with advanced capabilities in simulation of remodeling of soft tissue. The formulation is flexible so that its parameters could be adjusted to represent the arterial wall either as a single material or a material composed of several layers in finite element analyses of arteries. Explicit algorithms for both the material subroutine and the explicit integration with dynamic relaxation of equations of motion using finite element method are given. To circumvent the slow convergence of the standard dynamic relaxation and small time steps dictated by the stability of the explicit integrator, an adaptive dynamic relaxation technique that ensures stability and fastest possible convergence rates is developed. Incompressibility is enforced using penalty method with an updated penalty parameter. The model is used to simulate experimental data from the literature demonstrating that the model response is in excellent agreement with the data. An experimental procedure to determine the distribution of fiber directions in 3D for biological soft tissue is suggested in accordance with the microplane concept. It is also argued that this microplane formulation could be modified or extended to model many other phenomena of interest in biomechanics. PMID- 16706592 TI - Mechanical characterization of anisotropic planar biological soft tissues using large indentation: a computational feasibility study. AB - Traditionally, the complex mechanical behavior of planar soft biological tissues is characterized by (multi)axial tensile testing. While uniaxial tests do not provide sufficient information for a full characterization of the material anisotropy, biaxial tensile tests are difficult to perform and tethering effects limit the analyses to a small central portion of the test sample. In both cases, determination of local mechanical properties is not trivial. Local mechanical characterization may be performed by indentation testing. Conventional indentation tests, however, often assume linear elastic and isotropic material properties, and therefore these tests are of limited use in characterizing the nonlinear, anisotropic material behavior typical for planar soft biological tissues. In this study, a spherical indentation experiment assuming large deformations is proposed. A finite element model of the aortic valve leaflet demonstrates that combining force and deformation gradient data, one single indentation test provides sufficient information to characterize the local material behavior. Parameter estimation is used to fit the computational model to simulated experimental data. The aortic valve leaflet is chosen as a typical example. However, the proposed method is expected to apply for the mechanical characterization of planar soft biological materials in general. PMID- 16706593 TI - Lengthening of a single-loop tibialis tendon graft construct after cyclic loading: a study using Roentgen stereophotogrammetric analysis. AB - Although single-loop tibialis tendon allografts have increased in popularity owing to their many advantages over patellar tendon and double-loop hamstring tendon autografts, some percentage of the patient population do not have clinically stable knees following anterior cruciate ligament reconstruction with single-loop tibialis tendon allografts. Therefore, it would be advantageous to determine the causes of increased anterior laxity which ultimately must be traced to lengthening of the graft construct. One objective of this study was to demonstrate the feasibility of using Roentgen stereophotogrammetric analysis (RSA) to determine the causes of lengthening of a single-loop graft construct subjected to cyclic loading. A second objective was to determine which cause(s) contributes most to an increase in length of this graft construct. Radio-opaque markers were inserted into ten grafts to measure the lengthening at the sites of the tibial and femoral fixations and between the sites of fixation. Each graft was passed through a tibial tunnel in a calf tibia, looped around a rigid cross pin, and fixed to the tibia with a Washerloc fixation device. The grafts were cyclically loaded for 225,000 cycles from 20 to 170 N. Prior to and at intervals during the cyclic loading, simultaneous radiographs were taken. RSA was used to determine the three-dimensional coordinates of the markers from which the lengthening at the sites of fixation and between the sites of fixation was computed at each interval. The sites of the femoral and tibial fixations were the largest contributors to the increase in length of the graft construct, with maximum average values of 0.68 and 0.55 mm, respectively, after 225,000 cycles. The graft substance between the sites of fixation contributed least to lengthening of the graft, with a maximum average value of 0.31 mm. Ninety percent of the maximum average values occurred before 100,000 cycles of loading for the largest contributors. RSA proved to be a useful method for measuring lengthening due to all three causes. Lengthening of the graft construct at the sites of both fixations is sufficiently large that the combined contributions may manifest as a clinically important increase in anterior laxity. PMID- 16706594 TI - Oscillatory shear loading of bovine periodontal ligament--a methodological study. AB - This study examined the stress response of bovine periodontal ligament (PDL) under sinusoidal straining. The principle of the test consisted in subjecting transverse tooth, PDL and bone sections of known geometries to controlled oscillatory force application. The samples were secured to the actuator by support plates fabricated using a laser sintering technique to fit their contours to the tooth and the alveolar bone. The actuator was attached to the root slices located in the specimen's center. Hence the machine was able to push or pull the root relative to its surrounding alveolar bone. After determining an optimal distraction amplitude, the samples were cyclically loaded first in ramps and then in sinusoidal oscillations at frequencies ranging from 0.2 to 5 Hz. In the present study the following observations were made: (1) Imaging and the laser sintering technique can be used successfully to fabricate custom-made support plates for cross-sectional root-PDL-bone sections using a laser sintering technique, (2) the load-response curves were symmetric in the apical and the coronal directions, (3) both the stress response versus phase angle and the stress response versus. strain curves tended to "straighten" with increasing frequency, and (4) the phase lag between applied strain and resulting stress was small and did not differ in the intrusive and the extrusive directions. As no mechanical or time-dependent anisotropy was demonstrable in the intrusive and extrusive directions, such results may considerably simplify the development of constitutive laws for the PDL. PMID- 16706595 TI - A constituent-based model for the nonlinear viscoelastic behavior of ligaments. AB - This paper presents a constitutive model for predicting the nonlinear viscoelastic behavior of soft biological tissues and in particular of ligaments. The constitutive law is a generalization of the well-known quasi-linear viscoelastic theory (QLV) in which the elastic response of the tissue and the time-dependent properties are independently modeled and combined into a convolution time integral. The elastic behavior, based on the definition of anisotropic strain energy function, is extended to the time-dependent regime by means of a suitably developed time discretization scheme. The time-dependent constitutive law is based on the postulate that a constituent-based relaxation behavior may be defined through two different stress relaxation functions: one for the isotropic matrix and one for the reinforcing (collagen) fibers. The constitutive parameters of the viscoelastic model have been estimated by curve fitting the stress relaxation experiments conducted on medial collateral ligaments (MCLs) taken from the literature, whereas the predictive capability of the model was assessed by simulating experimental tests different from those used for the parameter estimation. In particular, creep tests at different maximum stresses have been successfully simulated. The proposed nonlinear viscoelastic model is able to predict the time-dependent response of ligaments described in experimental works (Bonifasi-Lista et al., 2005, J. Orthopaed. Res., 23, pp. 67 76; Hingorani et al., 2004, Ann. Biomed. Eng., 32, pp. 306-312; Provenzano et al., 2001, Ann. Biomed. Eng., 29, pp. 908-214; Weiss et al., 2002, J. Biomech., 35, pp. 943-950). In particular, the nonlinear viscoelastic response which implies different relaxation rates for different applied strains, as well as different creep rates for different applied stresses and direction-dependent relaxation behavior, can be described. PMID- 16706596 TI - Accumulation of E. Coli bacteria in mini-channel flow. AB - The objective of this research is to design and optimize a mini/micro-channel based surface-accumulator of E. coli bacteria to be detected by acoustic wave biosensors. A computational approach has been carried out using the state of the art software, CFD-ACE with water as bacteria bearing fluid. E. coli bacteria have been modeled as random discrete particles tracked by solving the Lagrangian equations. The design challenges are to achieve low shear force (pico-N), high concentration at accumulation, and high enough Reynolds number to avoid bacteria swimming. A range of low Reynolds number (Re) has been considered along with the effects of particle boundary interactions, gravity, Saffman lift, etc. More than two orders of magnitude higher concentration at the accumulation than the inlet concentration, and lower shear force of less than pico-N have been achieved in the optimized designs. PMID- 16706597 TI - Metabolic model of autoregulation in the Circle of Willis. AB - The Circle of Willis (CoW) is a ringlike structure of blood vessels found at the base of the brain. Its main function is to distribute oxygen-rich arterial blood to the cerebral mass. In a previous study, a one-dimensional (1D) model of the CoW was created to simulate a series of possible clinical scenarios such as occlusions in afferent arteries, absent or stringlike circulus vessels, or arterial infarctions (Moorhead et al., 2004, Comput. Methods Biomech. Biomed. Eng., 7(3), pp. 121-130). The model captured cerebral haemodynamic autoregulation by using a proportional-integral-derivative (PID) controller to modify efferent artery resistances. Although some good results and correlations were achieved, the model was too simple to capture all the transient dynamics of autoregulation. Hence a more physiologically accurate model has been created that additionally includes the oxygen dynamics that drive the autoregulatory response. Results very closely match accepted physiological response and limited clinical data. In addition, a set of boundary conditions and geometry is presented for which the autoregulated system cannot provide sufficient perfusion, representing a condition with increased risk of stroke and highlighting the importance of modeling the haemodynamics of the CoW. The system model created is computationally simple so it can be used to identify at-risk cerebral arterial geometries and conditions prior to surgery or other clinical procedures. PMID- 16706598 TI - Prediction of femoral head collapse in osteonecrosis. AB - The femoral head deteriorates in osteonecrosis. As a consequence of that, the cortical shell of the femoral head can buckle into the cancellous bone supporting it. In order to examine the buckling scenario we performed numerical analysis of a realistic femoral head model. The analysis included a solution of the hip contact problem, which provided the contact pressure distribution, and subsequent buckling simulation based on the given contact pressure. The contact problem was solved iteratively by approximating the cartilage by a discrete set of unilateral linear springs. The buckling calculations were based on a finite element mesh with brick elements for the cancellous bone and shell elements for the cortical shell. Results of 144 simulations for a variety of geometrical, material, and loading parameters strengthen the buckling scenario. They, particularly, show that the normal cancellous bone serves as a strong supporting foundation for the cortical shell and prevents it from buckling. However, under the development of osteonecrosis the deteriorating cancellous bone is unable to prevent the cortical shell from buckling and the critical pressure decreases with the decreasing Young modulus of the cancellous bone. The local buckling of the cortical shell seems to be the driving force of the progressive fracturing of the femoral head leading to its entire collapse. The buckling analysis provides an additional criterion of the femoral head collapse, the critical contact pressure. The buckling scenario also suggests a new argument in speculating on the femoral head reinforcement. If the entire collapse of the femoral head starts with the buckling of the cortical shell then it is reasonable to place the reinforcement as close to the cortical shell as possible. PMID- 16706599 TI - Towards a functional dissection of thioredoxin networks in plant cells. AB - Thioredoxins are a ubiquitous family of redox equivalent mediators, long considered to possess a limited number of target enzymes. Recent progress in proteomic research has allowed the identification of a wide variety of candidate proteins with which this small protein may interact in vivo. Moreover, the activity of thioredoxin itself has been recently found to be subject to regulation by posttranslational modifications, adding an additional level of complexity to the function of this intriguing enzyme family. The current review charts the technical progress made in the continuing discovery of the numerous and diverse roles played by these proteins in the regulation of redox networks in plant cells. PMID- 16706600 TI - Molecular mechanisms characterizing cone photoresponses. AB - In the vertebrate retina, rods mediate twilight vision and cones mediate daylight vision. Their photoresponse characteristics are different. The light-sensitivity of a cone is 10(2)-10(3) times lower than that of a rod. In addition, the photoresponse time course is much faster in cones. The mechanism characterizing cone photoresponses has not been known mainly because of the difficulty in isolating cones in large quantities to perform biochemistry. Recently, we developed a method to purify cones from carp retina using a density gradient, which made it possible to analyze the differences in the molecular mechanism of phototransduction between rods and cones. The results showed that signal amplification in cones is less effective, which explains the lower light sensitivity of cones. The results also showed that visual pigment phosphorylation, a quenching mechanism of light-activated visual pigment, is much more rapid in cones than in rods. The rapid phosphorylation in cones is attributed to a very high total kinase activity in cones. Because of this high activity, cone pigment is readily phosphorylated even at very high bleaching levels, which probably explains why cone photoresponses recover quickly. Based on these findings, the molecular mechanisms of the differences in the photoresponse characteristics between rods and cones are outlined. PMID- 16706601 TI - Two-photon singlet oxygen microscopy: the challenges of working with single cells. AB - A microscope is described in which singlet molecular oxygen, O2(a1deltag), is produced in a femtoliter focal volume via a nonlinear two-photon photosensitized process, and the 1270 nm phosphorescence from this population of O2(a1deltag) is detected in a photon counting experiment. Although two-photon excitation of a sensitizer is less efficient than excitation by a one-photon process, nonlinear excitation has several distinct advantages with respect to the spatial resolution accessible. Pertinent aspects of this two-photon O2(a1deltag) microscope were characterized using bulk solutions of photosensitizers. These data were compared to those obtained from a single biological cell upon linear one-photon excitation of a sensitizer incorporated in the cell. On the basis of the results obtained, we outline the challenges of using nonlinear optical techniques to create O2(aldeltag) at the single cell level and to then optically detect the O2(aldeltag) thus produced in a time-resolved experiment. PMID- 16706602 TI - Optimising the therapeutic safety of human embryonic stem cells through the evolution of defined culture systems supporting their isolation. AB - Realising the therapeutic promise of embryo-derived stem cells is absolutely dependent on the evolution of defined and quality assured in vitro culture systems to mediate their isolation, expansion and delivery. Critical to the safety of these systems, both for the transplant recipient and the general population, is the necessity to eliminate dependence on animal cell-derived reagents to reduce the risk of cross-specific disease transmission. This paper discusses progress in the development of defined culture systems for human embryonic stem cell isolation and the challenges that still must be overcome. PMID- 16706603 TI - Umbilical cord stem cell transplantation for primary immunodeficiencies. AB - Primary immunodeficiencies (PIDs) are a rare but important cause of mortality and morbidity in childhood: the most severe--known as severe combined immunodeficiency (SCID)--are fatal within the first year of life; other PIDs are less immediately life-threatening, but have a poor long-term outlook. Haematopoietic stem cell transplantation (HSCT) is the best treatment for SCID and is increasingly offered for other PIDs. The best results are achieved with an HLA-matched family donor. Umbilical cord stem cells (UCSCs) are an alternative stem cell source. Results using UCSCs in the treatment of haematological disorders and malignancy are as good as those for which marrow is the stem cell source. Although PIDs make up a small proportion of disorders amenable to treatment by HSCT, UCSCs are an ideal source of haematopoietic stem cells for many of these patients. Of the 52 patients with SCID or other PIDs for whom detailed information on outcome is available, results of engraftment, immune reconstitution, incidence of graft-versus-host disease and survival are comparable with other stem cell sources. Small stem cell dose and prolonged time to viral immunity limit the patients for whom UCSCs can be used. Newer methods of achieving better engraftment, ex vivo expansion of stem cells and generation of antigen-specific cytotoxic T cells are being developed at present, and will widen the application of UCSCs as a viable source for more patients. PMID- 16706604 TI - Adipose-derived stem cells for the regeneration of damaged tissues. AB - As the promise of stem cell-based therapies begins to be realised, and efforts to bring advances to the clinic mount, the source of these cells is increasingly important. The morbidity associated with harvesting stem cells from solid organs and the invasive nature of bone marrow biopsies may limit their practicality for wider clinical applications. An emerging body of literature suggests that adipose tissue may provide an abundant, readily accessible source of cells with similar potential to that described of other adult stem cells. This review will address advances in the use of adipose stem cells in fields as divergent as soft tissue reconstruction and cerebral infarction recovery. Numerous challenges will also be discussed; however, rapidly accumulating advances suggest that adipose stem cells may be as effective as they are abundant. PMID- 16706605 TI - Heat-shock protein 60 as a tool for novel therapeutic strategies that target the induction of regulatory T cells in human arthritis. AB - In health, immune responses to self are abundantly available, but under strict control of mechanisms of peripheral tolerance. Occasionally the immune system loses control and an autoimmune disease develops. At present, treatment of autoimmune disease is based on generalised suppression of all immune responses, and is often needed to be lifelong, leading to long-term toxicities and suppression of protective immune responses against pathogens. A more targeted approach would be to reset the immune system via restoration of failing regulatory mechanisms, and redirect the immune system to a state of tolerance. Over the past decade there have been enormous advances in the understanding of basic processes that control immune tolerance, pushing regulatory T cells forward as targets for novel therapeutic strategies. This review describes the development of antigen-specific immunotherapy that targets the antigen-specific induction of regulatory T cells as a means to treat autoimmune disease. The 'holy grail' for autoimmunity is not the disease-causing antigen, but the disease curing antigen. PMID- 16706606 TI - Dendritic cell immunotherapy for breast cancer. AB - Novel adjuvant therapies are urgently needed to complement the existing treatment options for breast cancer. The advent of the use of dendritic cells (DCs) for cancer immunotherapy provides a unique opportunity to overcome the relative non immunogenic property of breast tumours and address the underlying immunodeficiency. To date, the success of this approach has been limited, possibly due to the targeting of specific tumour antigens that rapidly mutate and, thus, become undetectable to the immune system. A more efficient approach would include preparations encompassing multiple antigens, such as those provided by loading of whole tumour cells or tumour RNA. It is proposed that targeting mammary stem cells responsible for resistance to chemo/immunotherapy, through the expression of a broad array of wild-type and mutated tumour antigens in the context of DCs, will become a mainstay for immunotherapy of breast cancer. PMID- 16706607 TI - Molecular biology of epidermal growth factor receptor inhibition for cancer therapy. AB - Understanding the role of the epidermal growth factor receptor (EGFR) in cellular signalling processes underlying malignancy has enabled the development of rationally designed EGFR-targeted therapeutics. Strategies have been devised to interfere with the EGFR signalling at three different levels: at the extracellular level, competing with ligand binding; at the intracellular level, inhibiting the activation of the tyrosine kinase; or at the mRNA level, modulating the expression of the EGFR protein. Each of these strategies has proven to have an antitumour effect mediated by events such as inhibition of cell proliferation, induction of apoptosis, decrease of cellular invasion and migration; and/or inhibition of angiogenesis. Furthermore, the combination of these strategies with traditional chemotherapy or radiotherapy has generally resulted in enhanced antitumour effects. Likewise, the benefit of interfering simultaneously with different signalling pathways has been documented to improve tumour growth inhibition. These preclinical results have encouraged clinical studies that led to the FDA approval of three drugs. However, finding the perfect strategy for each individual patient appears to be a limiting factor, demanding further research to be able to generate relevant molecular expression profiles on a case-to-case basis. Taken together, a successful EGFR inhibition will require a better understanding of signalling pathways in combination with the development of rationally designed effective molecules. PMID- 16706608 TI - Strategies to improve non-viral vectors--potential applications in clinical transplantation. AB - Prevention of acute rejection has been well controlled with immunosuppressive drugs. However, the long-term control of rejection is less satisfactory and the side effects of chronic usage of these drugs are far from acceptable. Thus, more imaginative options for therapy need to be explored. Gene therapy has potential promise in preserving allografts, preventing rejection and inducing tolerance. Despite this initial promise in many animal models, the translation of gene therapy to the clinical arena has been slow. This may be related in part to the deficiencies in vector development. Existing viral vectors are efficient at transducing allografts, but they induce inflammatory and pathogenic effects. Although the alternative non-viral systems are relatively innocuous, they are less efficient at gene delivery. This review systematically analyses the limitations of non-viral vector technology and the strategies that have been developed to overcome these limitations. Future development of non-viral vectors may have potential application in clinical transplantation. PMID- 16706609 TI - 9th Annual Meeting of the Regional Cancer Consortium for the Biological Therapy of Cancer. 16-18 February 2006, UPMC Herberman Conference Center, Pittsburgh, PA, USA. PMID- 16706613 TI - Characterization of envelope glycoprotein gp41 genotype and phenotypic susceptibility to enfuvirtide at baseline and on treatment in the phase III clinical trials TORO-1 and TORO-2. AB - Enfuvirtide (T-20) is the first entry inhibitor approved for treatment of HIV infection and acts by inhibiting conformational changes in the viral envelope protein gp41 that are necessary for fusion of the virus and host cell membranes. Here we present genotypic and phenotypic data on viral envelopes obtained at baseline (n = 627) and after 48 weeks of enfuvirtide treatment (n = 302) from patients in the TORO (T-20 versus Optimized Regimen Only)-1 and -2 phase III pivotal studies. The amino acid sequence at residues 36-45 of gp41 was highly conserved at baseline except for polymorphism of approximately 16% at position 42. Substitutions within gp41 residues 36-45 on treatment were observed in virus from 92.7% of patients who met protocol defined virological failure criteria and occurred in nearly all cases (98.8%) when decreases in susceptibility to enfuvirtide from baseline of greater than 4-fold were observed. Consistent with previous observations, a wide range of baseline susceptibilities (spanning 3 logs) was observed; however, lower in vitro baseline susceptibility was not significantly associated with a decreased virological response in vivo. Virological response was also independent of baseline coreceptor tropism and viral subtype. PMID- 16706614 TI - Highly active antiretroviral therapy reduces the age-associated risk of dementia in a cohort of older HIV-1-infected patients. AB - Historically, older patients have shown a higher risk of HIV-1-associated dementia (HIVD). The objective of this study was to evaluate the association of aging with HIVD and minor cognitive motor disorders (MCMDs) during the late highly active antiretroviral therapy (HAART) era and to analyze characteristics, predictive factors, and survival of older HIV-1-infected individuals affected by these disorders. A nested longitudinal study was designed for a cohort of HIV-1 infected individuals with neurological diseases. Multiple logistic regression and Cox regression for survival were employed. From 2000 to 2003, 195 patients with HIVD (53%) or MCMD (47%) were enrolled. The cumulative prevalence of these two disorders was 21%, with an increasing rate for calendar year (p < 0.001). Previous antiretroviral exposure occurred in 46% of patients. Mean CD4(+) cell count and plasma HIV-1 RNA were 144 cells/microl and 4.5 log10 copies/ml, respectively. The mean age was 44 years (SD, 9.9), with 35% of patients aged 20 39 years (I), 45% aged 40-49 years (II), and 20% aged >/=50 years (III). Among drug-naive patients, the prevalence of HIVD progressively increased in older subjects: 7.2% (I), 15.3% (II), and 27.3% (III) (p < 0.001), whereas no significant increase in HIVD with older age was observed in drug-treated subjects. Older age was independently associated with an increased risk of HIVD (odds ratio, 6.44; 95% confidence interval, 2.82-14.69) in naive but not in experienced individuals, but had no significant effect on survival. No significant effect of age was observed for MCMD. We conclude that in our cohort, HAART seems to alter the relationship between aging and HIVD, conferring a neuroprotective effect to older patients. These results may have significant implications for the clinical management of the older HIV population. PMID- 16706615 TI - Poor response to hepatitis C virus (HCV) therapy in HIV- and HCV-coinfected patients is not due to lower adherence to treatment. AB - The degree of adherence to anti-hepatitis C virus (HCV) therapy among HIV/HCV coinfected patients is not known. A prospective cohort study was performed in two groups of patients: 79 HIV/HCV-coinfected patients (group 1) and 78-HCV monoinfected patients (group 2). Patients were treated with interferon alpha-2a (3 million international units [MIU], three times per week) plus ribavirin (1000 1200 mg/day) for 48 weeks. Adherence to therapy was defined as having received +/ 80% of both drug dosages for +/-80% of the expected duration of therapy. The degree of adherence to treatment was similar for patients with or without HIV coinfection (72.2 versus 80.8%). The overall sustained virological response (SVR) in patients with adherence to therapy was 41.7% as compared with only 8.1% (p = 0.0001) in patients without adherence. The difference in SVR rate according to adherence to treatment was also evident in patients of group 1 (29.8% versus 9.1%; p = 0.05) as well as in those of group 2 (52.4 versus 6.7%; p = 0.001). Adherence to anti-HCV therapy, which can be similar in mono- and coinfected patients, enhances the likelihood of achieving an increase in SVR rate. In addition to improved adherence, in coinfected patients more aggressive therapeutic strategies may be necessary to achieve SVR. PMID- 16706616 TI - Silencing of HIV-1 subtype C primary isolates by expressed small hairpin RNAs targeted to gag. AB - Discovery of sequence-specific silencing by activating the RNA interference (RNAi) pathway has led to exciting new strategies for treating infection with human immunodeficiency virus type 1 (HIV-1). Of the HIV-1 subtypes, C is especially common in areas of the world that are worst affected. Although prone to mutation, genome plasticity of this subtype is limited in functionally important regions. We identified conserved sequences within the HIV-1 subtype C gag open reading frame and assessed whether they are suitable targets for inhibition of viral replication by RNA Pol III-driven small hairpin RNAs (shRNAs). Initially, the efficacy of each of a panel of 10 shRNAs against HIV-1 was determined using a reporter assay. shRNAs A and B, which targeted the 5 end of gag, were most effective and were used to assess inhibition of replication in cultured cells of two R5 isolates (Du151 and Du422) and one X4 virus (SW7). These shRNAs diminished intracellular HIV-1 gag RNA and HIV-1 protein concentrations as well as p24 secretion by up to 80% without inducing an interferon response. However, shRNA-mediated knockdown efficacy against each of these viral isolates varied slightly. These data show successful activation of RNAi to inhibit the replication of biologically distinct HIV-1 subtype C isolates. The effector shRNAs described here are potential candidates for gene therapy applications against the most common global subtype of HIV-1. PMID- 16706618 TI - Use of frozen-thawed cervical tissues in the organ culture system to measure anti HIV activities of candidate microbicides. AB - Cervical tissue-based organ culture system has been used to test the cytotoxicity and antiviral activity of microbicides. One of the problems of using current organ culture methods for routine microbicide testing is the need to continually obtain fresh tissue, which can be limited in access and supply. Use of frozen tissue, stored when available and thawed when needed, would alleviate the need for constant access to new tissue. This study was designed to explore the possibility of using frozen-thawed cervical tissue to test microbicides for their anti-HIV activity. We provided biochemical, histological, and quantitative immunohistochemical data to demonstrate the integrity of the frozen-thawed organ culture system. Significant levels of HIV-1 mucosal transmission were noted with both fresh and frozen-thawed tissue, regardless of the coreceptor usage of the virus isolate. Furthermore, candidate microbicides UC781, beta-cyclodextrin, and octylglycerol inhibited HIV-1 transmission across the mucosa of frozen-thawed tissues with a level of efficiency similar to that of fresh tissues. Therefore, frozen-thawed cervical tissue in the organ culture system provides a practical and convenient model to screen topical microbicides for their ability to block sexual transmission of HIV-1, and reduces the problems associated with procurement of the numerous tissues required for evaluation and comparison of microbicide candidates among different laboratories. PMID- 16706617 TI - Cellulose acetate 1,2-benzenedicarboxylate inhibits infection by cell-free and cell-associated primary HIV-1 isolates. AB - Cellulose acetate 1,2-benzenedicarboxylate (CAP), a pharmaceutical excipient used for enteric film coating of capsules and tablets, was previously shown to have potent inhibitory activity against infection by human immunodeficiency virus type 1 (HIV-1) T cell line-adapted (TCLA) strains. In the present study, we determined the inhibitory activity of CAP against infection by cell-free and cell-associated primary HIV-1 isolates with distinct genotypes and biotypes in cervical explants, peripheral blood mononuclear cells (PBMCs), monocytederived macrophages (MDMs), and CEMx174 5.25M7 cells. CAP blocked infection by cell-free and cell-associated HIV-1 in cervical explants. It inhibited infection by cell-free primary HIV-1 isolates (clades A to G and group O) in PBMCs, MDMs, and CEMx174 5.25M7 cells and blocked transmissions of the cell-associated primary HIV-1 isolates from dendritic cells (DCs) to PBMCs, from MDMs to PBMCs, and from PBMCs to CEMx174 5.25M7 cells. The inhibitory activity of CAP on infection by the cell-free and cell-associated primary HIV-1 isolates is independent of viral subtypes and coreceptor usage. These data suggest that CAP is a good microbicide candidate that can be further developed for preventing sexual transmission of HIV-1. PMID- 16706619 TI - HIV-1 infection downregulates nuclear telomerase activity on lymphoblastoic cells without affecting the enzymatic components at the transcriptional level. AB - Both increased lymphocyte renewal with subsequent exhaustion of the immune system and impaired T cell renewal have been put into view to account for CD4+ T cell depletion and development of AIDS in HIV-1- infected humans. Telomerase is an enzyme that is involved in mechanisms that control cell life span and replicative potential. The effect of HIV-1 on telomerase activity, certain regulators, and telomeric terminal restriction fragment length on lymphoid Jurkat cells was used in measuring the proliferative activity of T lymphoid cells before and after being infected. At the cellular level, the enzymatic activity remains almost stable but further analyses of fractionated cells revealed that telomerase activity in the nuclear compartment was diminished whereas in the cytoplasmic compartment it was relatively increased on HIV-1 infection. Two key components of telomerase regulation were further considered at the transcriptional level, that is, the mRNA levels of both human telomerase reverse transcriptase (hTERT)- including the relative amount of its alternative splicing variants--and hTR. They were unaffected on HIV-1 infection. Telomeric length was also conserved in infected cells. Overall, these findings demonstrate that HIV-1 infection of Jurkat cells down modulate telomerase activity in the nuclear compartment by affecting its cellular localization. PMID- 16706621 TI - Real-time PCR quantitation of subtype C HIV DNA in a Zambian discordant couple cohort. AB - To study the viral sequence diversity that is characteristic of HIV infection, PCR amplification and sequencing of viral genes is an essential step. However, a limitation of traditional PCR methods is that one viral target may be preferentially amplified over another when multiple sequences are present. This presents a particular problem when conclusions about diversity are made from one or only a few PCRs. One way to avoid resampling is to perform a large number of PCR amplifications on a single template; however, this requires that extensive dilution series be carried out on each patient sample to identify the appropriate concentration of input DNA. Here we describe the development and implementation of a quantitative real-time PCR (qPCR) method that detects a short sequence in gag and is optimized to detect subtype C HIV sequences. The standard curve was externally validated using two chronically infected cell lines carrying a known number of HIV copies per genome, and this assay yielded reproducible and accurate measurements on patient DNA samples over a wide range of input targets. The qPCR assay results were consistent with those obtained by the traditional limiting dilution method yet entailed only a fraction of the time and reagents required for the latter. This robust and quantitative real-time assay can be used to ensure that each viral sequence obtained through PCR represents a single template for studies in which the diversity of the entire population must be accurately portrayed, and can readily be applied to other research settings and viral subtypes. PMID- 16706622 TI - Expansion after epitope peptide exposure in vitro predicts cytotoxic T lymphocyte epitope dominance hierarchy in lymphocytes of vaccinated mamu-a*01+ rhesus monkeys. AB - Because of the importance of developing HIV vaccine strategies that generate cytotoxic T lymphocyte (CTL) responses with a maximal breadth of epitope recognition, we have explored a variety of novel strategies designed to overcome the usual propensity of CTLs to focus recognition on a limited number of dominant epitopes. In studies of rhesus monkeys expressing the Mamu-A*01 MHC class I allele, we show that variously configured multiepitope plasmid DNA vaccine constructs elicit CTL populations that do not evidence skewing of recognition to dominant epitopes. Nevertheless, repeated boosting of these vaccinated monkeys with different live recombinant vaccine vectors uncovers and amplifies the usual CTL epitope dominance hierarchy. Importantly, in vitro peptide stimulation of peripheral blood mononuclear cells from monkeys that have received only a multiepitope plasmid DNA priming immunization uncovers this dominance hierarchy. Therefore, the dominance hierarchy of the vaccine-elicited epitope-specific CTL populations is inherent in the T lymphocytes of the monkeys after initial exposure to epitope peptides, and the ultimate breadth of epitope recognition cannot be modified thereafter. This finding underscores the enormous challenge associated with increasing the breadth of CTL recognition through vaccination. PMID- 16706620 TI - Diversity in HIV-1 envelope V1-V3 sequences early in infection reflects sequence diversity throughout the HIV-1 genome but does not predict the extent of sequence diversity during chronic infection. AB - Differences in the extent of genetic diversity have been observed in human immunodeficiency virus type-1 (HIV-1) envelope sequences early in infection, and this has been linked to gender and to modifiable exogenous factors such as hormonal contraceptive use and genital tract infections. But it is unclear whether envelope diversity is indicative of diversity in other regions of the viral genome, and thus whether it adequately reflects whether multiple or a single virus initiated the infection. Here we show that six women with homogeneous envelope V1-V3 sequences during primary infection also had homogeneous gag and polymerase (pol) sequences at the same time. On the other hand, six women with multiple envelope sequences had diverse gag and pol genotypes during a similar interval after infection. This suggests that envelope sequences reflect sequence diversity throughout the viral genomes present early in infection and thus provide an indication of whether a single virus or multiple viruses initiated the infection. Analysis of HIV-1 sequences from about 3 years after infection revealed that the level of diversity and diversification was similar between the women in the two groups. PMID- 16706623 TI - HIV-1 Group N: evidence of ongoing transmission in Cameroon. AB - An HIV-1 group N infection, 02CM-DJO0135, was identified among specimens collected in 2002 at the D'Joungolo Hospital, Yaounde, Cameroon. Sequences were obtained from viral RNA extracted from plasma for regions of LTR-gag, pol-vif, and env. The virus amplified from the specimen is closely related to a previously reported group N virus, 02CM-DJO0131, that was also collected at this hospital in 2002. Although the viral sequences for the two isolates differ, their close relationship suggests that the two specimens are linked. No patient histories are available for 02CM-DJO0131 and 02CM-DJO0135; the specimens could have been drawn from a husband/wife, mother/child, or a single individual. However, differences in seroreactivity indicate that it is unlikely that the specimens were drawn from the same patient. This report documents the second case that suggests linkage between group N-infected individuals and indicates that there is ongoing transmission of HIV-1 group N in Cameroon. PMID- 16706624 TI - Genotypic and phenotypic characterization of viral isolates from HIV-1 subtype C infected children with slow and rapid disease progression. AB - The genotypes and biological phenotypes of HIV-1 isolates obtained from 40 perinatally infected children in South Africa were analyzed. This included 15 infants who had HIV-related symptoms, most of whom died within 2 years of birth (rapid progressors), and 25 children who survived between 4 and 9 years with varying signs of disease (slow progressors). Heteroduplex mobility assays and sequence analysis confirmed that within the env and gag regions, all isolates were HIV-1 subtype C. Viral isolates from 14 of the 15 rapid progressors used the CCR5 coreceptor, whereas 1 (02ZARP1) used both the CXCR4 and CCR5 coreceptors. Among the 25 slow progressors, 22 isolates used CCR5 only, 2 used CXCR4 only, and 1 used both CCR5 and CXCR4. Two of the slow-progressing children who harbored CXCR4-using viruses had AIDS. All four CXCR4-using viruses had genotypic changes in the V3 region previously shown to be associated with CXCR4 usage. This cross sectional study shows that HIV-1 subtype C viruses from both rapid- and slow progressing perinatally infected children used predominantly CCR5. Similar to adults, CXCR4 usage was uncommon among HIV-1 subtype C isolates from pediatric infections. PMID- 16706625 TI - HTLV-1 p12I protein sequences from South America: truncated proteins and common genetic signatures. AB - p12 pX ORF-I (p12I) of simian T cell lymphotropic virus 1 from Central and West Africa is a 91-amino acid (aa) protein, contrasting with the known p12 sequences from human T cell lymphotropic virus type 1, which are 99 aa in length. An in silico sequence analysis shows a premature termination codon in HTLV-1 p12I sequences from South American and Japanese strains. A translation analysis indicated the presence of 86-aa and 81-aa p12I proteins, shorter than the 99-aa protein found in the majority of HTLV-1 strains around the world. This report shows one more common feature between these two viruses. Additionally, the HTLV-1 p12 signature reported as a genetic marker of Brazilian sequences (p12I P63 and P91) is also highly prevalent in p12 Argentinean sequences. Because p12I may confer a proliferative advantage on HTLV-1-infected cells the existence of naturally truncated proteins might supply insights into its pathogenic mechanisms. PMID- 16706626 TI - HIV-1 genetic diversity and genotypic drug susceptibility in the Republic of Georgia. AB - The genetic diversity and genotypic drug susceptibility of HIV-1 strains circulating in the Republic of Georgia, formerly part of the Soviet Union, were investigated for first time. Forty-eight HIV-positive drug-naive Georgian individuals contributed PBMC DNA between 1998 and 2003. On the basis of phylogenetic analyses of partial pol sequences, the predominant HIV-1 genetic forms were subtype A (70%), followed by subtype B (26%); both genetic forms were carried by injecting drug users and heterosexuals. There was also one subtype C (2%) and one CRF18_cpx (2%). The Georgian subtype A strains clustered with subtype A from Russia, designated A(FSU). Twelve of the subtype A strains (25%) contained the secondary protease inhibitor mutation V77I and 9 also had two other silent mutations. This "V77I haplotype" marks one particular genetic lineage of the epidemic in the former Soviet Union. Two strains (4%) carried antiretroviral (ARV) drug resistance mutations. Nearly full-length genome sequences of five Georgian strains were also completed. Two, 98GEMZ011 (subtype A) and 98GEMZ003 (subtype B), closely resembled the parental strains that recombined to create CRF03_AB. The use of these parental strains in the analysis revealed an additional segment of subtype A in CRF03_AB. Thus, the HIV-1 epidemic in Georgia was composed of a mixture of subtype A(FSU) and subtype B. PMID- 16706627 TI - Editorial: what ails solid tumor radioimmunotherapy? PMID- 16706628 TI - Editorial: "right place, wrong place": extravasation of therapeutic drug for molecular targeted radiotherapy. PMID- 16706629 TI - A phase I trial of (90)Y-DOTA-anti-CEA chimeric T84.66 (cT84.66) radioimmunotherapy in patients with metastatic CEA-producing malignancies. AB - PURPOSE/OBJECTIVE: Previous radioimmunotherapy (RIT) clinical trials at this institution with (90)Y-labeled cT84.66 anti-CEA (carcinoembryonic antigen) evaluated the antibody conjugated to diethylenetriaminepentaacetic acid (DTPA). The aim of this phase I therapy trial was to evaluate cT84.66 conjugated to the macrocyclic chelate (90)Y-DOTA and labeled with (90)Y in a comparable patient population. EXPERIMENTAL DESIGN: Patients with metastatic CEA-producing cancers were entered in this trial. If antibody targeting to tumor was observed after the administration of (111)In-DTPA cT84.66, the patient then received the therapy infusion of (90)Y-DOTA-cT84.66 1 week later. Serial nuclear scans, blood and urine collections, and computed tomography (CT) scans were performed to assess antibody biodistribution, pharmacokinetics, toxicities, and antitumor effects. RESULTS: Thirteen (13) patients were treated in this study. Dose-limiting hematologic toxicity was experienced at initial starting activity levels of 12 and 8 mCi/m(2). Subsequent patients received systemic Ca-DTPA at 125 mg/m(2) every 12 hours for 3 days post-therapy to allow for a dose escalation to 16 mCi/m(2), where hematologic toxicity was observed with an associated maximum tolerated dose (MTD) of 13.4 mCi/m(2). Tumor doses ranged from 4.4 to 569 cGy/mCi, which translated to 97-12,500 cGy after a single infusion of (90)Y-DOTA cT84.66. Human anti-chimeric antibody (HACA) response developed in 8 of 13 patients and prevented additional therapy in 4 patients. CONCLUSIONS: This study demonstrates the feasibility of using (90)Y-DOTA-cT84.66 for antibody-guided radiation therapy. Immunogenicity of the DOTA-conjugated cT84.66 antibody was not appreciably greater than that observed with (90)Y-DTPA-cT84.66 in previous trials. Dose-limiting hematopoietic toxicity with (90)Y-DOTA-cT84.66 decreased with Ca-DTPA infusions post-therapy and appears to be comparable to previously published results for (90)Y-DTPA-cT84.66. The highest antibody uptake and tumor doses were to small nodal lesions, which supports the predictions from preclinical and clinical data that RIT may be best applied in the minimal tumor burden setting. PMID- 16706630 TI - Extravazation of therapeutic yttrium-90-ibritumomab tiuxetan (zevalin): a case report. AB - The occurrence of radiation necrosis to the extravasation of therapeutic radionuclide-yttrium-90-ibritumomab tiuxetan-is described, as well as the subsequent steps taken to both address the occurrence and to prevent the re occurrence of such an adverse event. Published reports on the extravasation of diagnostic and therapeutic radionuclides are reviewed, as are prevention and treatment protocols. PMID- 16706631 TI - Pharmacokinetics and tumor localization of (111)in-labeled HuCC49DeltaC(H)2 in BALB/c mice and athymic murine colon carcinoma xenograft. AB - The primary limitation of IgG antibodies for radioimmunotherapy of solid tumors is their prolonged serum half-life, leading to dose-limiting bone marrow toxicity at doses providing inadequate radiation to the tumor. A humanized C(H)2 domain deleted variant of the anti-TAG-72 antibody CC49 (HuCC49DeltaC(H)2) has faster blood clearance, compared to the IgG, while retaining tumor targeting. We compared the pharmacokinetics and tumor uptake of (111)In-HuCC49DeltaC(H)2 in BALB/c mice and a colon carcinoma (LS-174T) mouse xenograft with that of (111)In labeled chimeric CC49 (cCC49), an antibody with pharmacokinetics similar to the humanized CC49 parent. Immuno-conjugates of HuCC49DeltaC(H)2 and cCC49 prepared with the (111)In chelator Mx-DTPA (1-isothiocyantobenzyl-3 methyldiethylenetriaminepentaacetic acid) retained low nM affinity and radiolabeling protocols provided greater than 95% radio-incorporation with (111)In while retaining greater than 80% immunoreactivity. Blood clearance of (111)In-HuCC49DeltaC(H)2 in BALB/c mice was monoexponential (t(1/2) 5.4 hours) and faster than (111)In-cCC49 (biexponential clearance; t1/2Delta 1.5 hours; t1/2beta 162 hours). The (111)In-HuCC49DeltaC(H)2 also cleared more rapidly from the blood in the murine xenograft. At 1 hour postinjection, blood concentrations for (111)In-HuCC49DeltaC(H)2 and (111)In-cCC49 were comparable (25.5 injected dose per g [%ID/g] and 21.3 %ID/g, respectively); tumor uptake for (111)In- HuCC49DeltaC(H)2 was 7.9 %ID/g, compared to 7.5 %ID/g for (111)In-cCC49. However, at 24 hours, blood concentration for (111)In-HuCC49DeltaC(H)2 was less than (111)In-cCC49 (0.9 %ID/g versus 5.2 %ID/g, respectively) with comparable tumor retention (14.4 %ID/g versus 19.0 %ID/g, respectively). Faster blood clearance of (111)In-HuCC49DeltaC(H)2 and tumor localization comparable to that of (111)In cCC49 provided a fourfold improved tumor-to-blood ratio for (111)In HuCC49DeltaC(H)2 at 24 hours postinjection. PMID- 16706632 TI - Radiolabeled melanin-binding peptides are safe and effective in treatment of human pigmented melanoma in a mouse model of disease. AB - The incidence of melanoma is rising, and therapeutic options for metastatic melanoma are limited. We report the results of experimental melanoma therapy with 188-Rhenium-labeled melanin-binding decapeptide ((188)RE-HYNIC-4B4) and a comprehensive safety evaluation of this treatment. (188)RE-HYNIC- 4B4 bound only to nonviable eumelanotic MNT1 and pheomelanotic SK-28-MEL human melanoma cells in vitro, as determined by immunofluorescence, which is consistent with the inaccessibility of intracellular melanin in live cells, and suggests specificity for tumors with a significant amount of extracellular melanin. Administration of 1 mCi (188)RE-HYNIC-4B4 to MNT1 tumor-bearing mice significantly slowed tumor growth, with the therapeutic effect being a result of specific binding to tumor melanin, as irrelevant (188)RE-labeled decapeptide did not produce therapeutic gain. Repeated doses of (188)RE-HYNIC-4B4 had a more profound effect on tumor growth than a single dose. Treatment of tumors with 0.3-0.4 cm diameter was more effective than of larger ones (0.5-0.7 cm). There was no difference in uptake of (188)REHYNIC- 4B4 in melanized tissues of black C57BL6 mice and no histologically apparent damage to these tissues in comparison with white BALB/C mice. Treatment of C57BL6 mice with (188)RE-HYNIC-4B4 did not change their behavior, as established by SHIRPA protocol, and did not cause damage to neurons and glial cells. These results indicate that radiolabeled melanin-binding peptides are efficient and safe in treatment of melanoma and could be potentially useful against this tumor. PMID- 16706633 TI - Phase II trial of subcutaneous interferon followed by intravenous hybrid bolus/continuous infusion interleukin-2 in the treatment of renal cell carcinoma: final results of Cancer Biotherapy Research Group 95-09. AB - OBJECTIVE: We conducted a phase II trial in metastatic renal cell cancer of outpatient subcutaneous (s.c.) interferon-alpha2b (IFN), followed by an inpatient hybrid schedule of bolus and continuous interleukin-2 (IL- 2). METHODS: Treatment consisted of monthly IFN 10 MU/m(2) s.c. for 4 consecutive days, followed by 36 MIU/m(2) bolus IL-2, then 72-hour continuous intravenous (i.v.) infusion of 18 MIU/m(2) IL-2 per day. Between May 1997 and June 2000, 25 men and 11 women enrolled, with a median age of 57 years (range, 42-77), including 9 patients over 65. Prior treatment included nephrectomy (31), radiation (8), biotherapy (7), and chemotherapy (4). Sites of disease included 26 lung, 13 lymph node, 9 bone, 8 liver, 4 kidney, and 4 adrenal locations. Patients received an average of 3.1 treatment cycles (range, 1-6). RESULTS: There was 1 complete and 3 partial responses, for a response rate of 11% (3% to 27%; 95% confidence interval [CI]); 40% had stable disease. Median failure-free survival was 2.5 months; median overall survival was 15.0 months. The 1-, 2-, and 5-year survival rates were 53%, 30%, and 12%, respectively. Only 8 patients required a reduction in IL-2 dose. The most frequent grade 3 or 4 toxicities were 11% fatigue, 9% renal insufficiency, and 7% hypotension. CONCLUSIONS: Response and survival rates were similar to those seen in other multicenter trials using inpatient high-dose IL-2. PMID- 16706634 TI - Complete resolution of malignant ascites in stage IV breast cancer by peritoneal drainage and innovative chemoimmunotherapy: a case report. AB - Malignant ascites is a very serious and difficult to manage problem in the cancer patient. It is a sign of late stage disease and treatment is mainly palliative. Therefore the treatment should produce minimal discomfort and have few side effects. Numerous methods for managing the problem have been attempted, but few have afforded complete remission of the ascites with improved survival. We are reporting a case of complete remission of malignant ascites in a stage IV breast cancer patient. Her condition was managed by a peritoneal dialysis catheter and an aggressive intraperitoneal innovative chemoimmunotherapy protocol. PMID- 16706635 TI - Intradermal vaccination of dendritic cell-derived exosomes is superior to a subcutaneous one in the induction of antitumor immunity. AB - Because dendritic cell (DC)-derived exosomes (EXO) harbor many important DC molecules involved in inducing immune responses, EXO-based vaccines have been extensively used to induce antitumor immunity in different animal tumor models. However, it is not clear which route of EXO administration can induce more efficient antitumor immune responses. In this study, we compared the antitumor immunity derived from EXO vaccine by way of the two common administration routes, the subcutaneous (s.c.) and the intradermal (i.d.) administrations. Our data showed that the i.d. EXO administration resulted in more EXO-absorbed DC migrating into the T-cell areas of draining lymph nodes than the s.c. administration. Interestingly, the i.d. EXO administration also resulted in an enhanced ovalbumin (OVA)-specific CD8(+) T-cell proliferation and CD8(+) CTL effector responses in vivo, compared to the s.c. administration. Similarly, compared to the s.c. vaccination, the i.d. vaccination induced stronger antitumor immunity in the animal tumor model. Therefore, the i.d. EXO vaccination is superior to the s.c. one and should be considered when EXO-based vaccine is designed. PMID- 16706637 TI - Application of I(125) brachytherapy combined artificial joint prosthesis in malignant osteo- and soft-tissue sarcoma. AB - OBJECTIVES: The aim of this study was to evaluate the brachytherapy effectiveness of I(125) seeds combined with artificial prosthesis in malignant sarcoma therapy. METHODS: The combination of I(125) seeds and artificial prosthesis was implanted to replace tumor section for three clinical cases-2 malignant osteosarcoma patients and 1 malignant soft-tissue tumor patient- through the direct operation. RESULTS: Approximately 14-18 months after the operation, the results of our post operational investigation showed that the tumor tissues of 3 patients had been completely removed. The limb functions recovered well. The brachytherapy of the combination of I(125) seeds and artificial prosthesis in malignant tumor improved the curative effect. No tumor existed, and no infection occurred. CONCLUSIONS: It was demonstrated that this method was safe and easy. No side-effect was observed after the implantation of I(125) seeds. The brachytherapy was proven to be a potential method for patients who were at high risk to recrudesce the malignant osteosarcoma of tumors after the tumor excision. PMID- 16706638 TI - Compression of morbidity: the hype and the reality, part 2. PMID- 16706636 TI - Evaluation of combined (177)Lu-DOTA-8-AOC-BBN (7-14)NH(2) GRP receptor-targeted radiotherapy and chemotherapy in PC-3 human prostate tumor cell xenografted SCID mice. AB - The focus of this study was to evaluate the therapeutic benefit of combined gastrin-releasing peptide (GRP) receptor-targeted radiotherapy (TRT) with chemotherapy, using the PC-3 xenograft severe combined immunodeficiency (SCID) mouse model. (177)Lu-DOTA-8-AOC-BBN(7-14)NH(2) is a radiotherapeutic peptide that specifically targets the gastrin-releasing peptide receptor overexpressed on primary and metastatic prostate cancer. The chemotherapeutic agents, docetaxel and estramustine, were administered as single agents or in combination with the receptor-targeted radiotherapeutic agent. Combination receptor TRT/chemotherapy studies were begun 21 days postxenografting and were conducted as multiple-dose trials. The GRP receptor TRT agent was administered every 14 days, and single and combination chemotherapy dose regimens were given weekly. Tumor size, body weight, and body condition score were evaluated twice-weekly and a hematology profile once-weekly. Therapy study tumor volumes were evaluated by way of a repeated measures analysis of variance (ANOVA). Tumor volume measurements at 12 days postdose administration demonstrated a statistically significant (two-tailed P-value <0.05) tumor growth suppression in all experimental groups receiving GRP receptor-targeted radiotherapy, when compared to the control group. The two combined GRP receptor TRT/chemotherapy treatment groups demonstrated the greatest tumor growth suppression of all treatment groups. In comparing the two combined GRP receptor TRT/chemotherapy groups to the GRP receptor TRT alone group, a statistically significant difference was demonstrated for the combined groups by day 30, postdose administration. These data demonstrate that GRP receptor targeted radiation therapy, using (177)Lu-DOTA-8-AOC-BBN(7-14)NH(2), used either alone or in combination with conventional chemotherapy, can suppress the growth of androgen- independent prostate cancer (AIPC). PMID- 16706639 TI - Extracellular redox state: refining the definition of oxidative stress in aging. AB - Oxidative stress in aging can result from an imbalance of prooxidants and antioxidants with excessive, destructive free radical chemistry. Thiol systems are important in the control of these processes, both by protecting against damage and serving in redox signaling mechanisms to sense danger and repair the damage. Studies by a number of research groups in collaboration with the Emory Clinical Biomarkers Laboratory show that the redox state of the central tissue antioxidant, glutathione (GSH), can be measured in human plasma and provides a quantitative systemic indicator of oxidative stress. Plasma GSH/GSSG redox in humans becomes oxidized with age, in response to chemotherapy, as a consequence of cigarette smoking, and in association with common age-related diseases (e.g., type 2 diabetes, cardiovascular disease). However, the GSH/GSSG redox is not equilibrated with the larger plasma cysteine/cystine (Cys/CySS) pool, and the Cys/CySS redox varies with age in a pattern that is distinct from that of GSH/GSSG redox. Furthermore, in vitro studies show that variation in Cys/CySS redox over the range found in vivo affects signaling pathways, which control cell proliferation and oxidant-induced apoptosis. The results point to the conclusion that free radical scavenging antioxidants are of increased importance when thiol/disulfide redox states are oxidized. Because thiol/disulfide redox states, per se, function in redox signaling and control as well as antioxidant protection, GSH/GSSG and Cys/CySS redox states may provide central parameters to link environmental influences and progression of changes associated with aging. PMID- 16706640 TI - Factors that might affect the allotopic replacement of a damaged mitochondrial DNA-encoded protein. AB - The human mitochondrion contains a small circular genome that codes for 13 proteins, 22 tRNAs, and 2 rRNAs. The proteins are all inner membrane bound components of complexes involved in the electron transport system and ATP formation. Mutations to any of the 13 proteins affect cellular behavior because energy production could be decreased. Investigators have attempted to find methods to correct these mutated proteins. One way is to express the mitochondrial gene in the nucleus (called allotopic expression). The newly synthesized protein would have to be imported into mitochondria and assembled into complexes. This paper reviews some of the successful attempts to achieve allotopic expression and discusses some issues that might affect the ability to have the proteins properly inserted into the inner membrane. PMID- 16706641 TI - Can a single subunit yeast NADH dehydrogenase (Ndi1) remedy diseases caused by respiratory complex I defects? AB - The proton-translocating NADH-quinone oxidoreductase (complex I) is one of five enzyme complexes in the oxidative phosphorylation system in mammalian mitochondria. Complex I is composed of 46 different subunits, 7 of which are encoded by mitochondrial DNA. Defects of complex I are involved in many human mitochondrial diseases; therefore, the authors proposed to use the NDI1 gene encoding a single subunit NADH dehydrogenase of Saccharomyces cerevisiae for repair of respiratory activity. The yeast NDI1 gene was successfully introduced into 10 mammalian cell lines (two of which were complex I-deficient mutants). The expressed Ndi1 protein was correctly targeted to the matrix side of the inner mitochondrial membranes, was fully functional, and restored the NADH oxidase activity to the complex I-deficient cells. The NDI1-transduced cells were more resistant to complex I inhibitors and diminished production of reactive oxygen species. It was further shown that the Ndi1 protein can be functionally expressed in tissues such as skeletal muscles and brain of rodents. The Ndi1 expression scarcely induced an inflammatory response as assessed by hematoxylin and eosin (H&E) staining. The Ndi1 protein expressed in the substantia nigra (SN) elicited protective effects against neurodegeneration caused by 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine treatment. The Ndi1 protein has a great potential as a molecular remedy for complex I deficiencies. PMID- 16706642 TI - Protective efficacy of alpha-lipoic acid on acetylcholinesterase activity in aged rat brain regions. AB - The purpose of the present investigation was to measure the activity of acetylcholinesterase in discrete regions of young and aged rat brain before and after DL-alpha-lipoic acid supplementation. Two groups of male albino rats were used in this study (4 and 24 months of age). DL-alpha-lipoic acid was administered intraperitoneally with a regimen of 100 mg/kg body weight per day using alkaline saline as a vehicle for 7 and 14 days. The activity was measured in the cerebral cortex, cerebellum, striatum, hippocampus and hypothalamus, and found to be significantly decreased in some of the brain regions in aged rats. Administration of lipoic acid into aged rats reversed the decrease in the activity in the discrete brain regions. These results suggest that lipoic acid is effective in restoration of the activity of acetylcholinesterase in aged rats. PMID- 16706643 TI - Cytochemical estimation of cytochrome oxidase activity as a morphofunctional mitochondrial check-up. AB - Cytochemical estimation of COX activity and morphometric measurement of mitochondrial ultrastructure were carried out in organelles from adult and old rats. Although no age-related difference was found in cytochemical precipitate (CPA) and mitochondrial area (MA), the ratio CPA:MA (R) decreased by 25.7% in aging. R was the same in oversized mitochondria, but in smaller organelles it was significantly decreased during aging. R reports on the functional mitochondrial surface involved in energy providing mechanisms; thus, by revealing age-related differences, these data suggest that the COX preferential cytochemistry associated with morphometry may serve as a reliable and sensitive mitochondrial morphofunctional checkup procedure. PMID- 16706644 TI - From disease-oriented to aging/longevity-oriented studies. AB - Aging should be considered a major risk factor for life-threatening degenerative pathologies including atherosclerosis, cancer, neurodegeneration, diabetes type II, osteoporosis, and sarcopenia. Although an apparent paradox, it appears that the most effective way to delay or even to avert age-related diseases is to live longer. Common changes in the epigenetic control of gene expression may be one of the central mechanisms behind both aging and age-associated pathologies. If so, epigenetic interventions may serve in a twofold manner: (a) to extend the lifespan and (b) cure age-related degenerative diseases. Currently predominant disease-oriented paradigm should be reconsidered toward aging/longevity oriented. PMID- 16706645 TI - Tissue-specific effect of age and caloric restriction diet on mitochondrial DNA content. AB - The effect of age and caloric-restriction (CR) diet on mitochondrial DNA (mtDNA) content in different rat tissues was investigated. A decrease of the mtDNA content occurs with aging in liver and soleus muscle, whereas there is no age related significant change of mtDNA content in brain. CR fully reverses the age dependent loss of mtDNA in liver and soleus, whereas it results in a significant increase of mtDNA amount above the value of aged ad libitum fed rats in brain. These results further support the tissue-specific effect of CR, likely because of the different dependence of tissues on external nutrient uptake. PMID- 16706646 TI - Testing mitochondrial metabolic competence by cytochrome oxidase preferential cytochemistry versus immunoreactivity of subunits I and IV. AB - Cytochemically evidenced COX activity was compared with levels of immunohistochemically stained mitochondrial- and nuclear-encoded subunits (CO I and CO IV) in the dentate gyrus outer molecular layer (OML) and cerebellar granular layer (GL) of adult and old rats. COX activity decreased significantly in aging, whereas CO I and CO IV levels were significantly increased both in GL and OML of old animals. These findings suggest that the age-related decay of the mitochondrial metabolic competence is not caused by a reduction of COX subunits levels, but causal events affecting mitochondria as discrete morphofunctional units of the cellular bioenergetic machinery. PMID- 16706647 TI - Caloric restriction protects mitochondrial function with aging in skeletal and cardiac muscles. AB - In skeletal muscles and heart in vitro complex IV activity is lower in young adult caloric restricted (CR) animals despite normal aerobic function in situ and in vivo. On the other hand, whereas markers of oxidative capacity decline 25% to 46% between 8 and 10 months and 35 months in ad libitum fed (AL) animals, in most muscles there is no decline in CR across the same absolute age (35 mo old) or relative age (35% survival rate) span and PGC-1alpha gene expression in gastrocnemius muscle declines more slowly with aging. The present results show that CR largely prevents the age-associated decline in mitochondrial function in heart and skeletal muscles, and suggest that this is secondary to a better maintained drive on mitochondrial biogenesis. PMID- 16706648 TI - Mitochondrial genome anatomy and species-specific lifespan. AB - Several lines of evidence implicate mtDNA in the mechanisms of aging and longevity. The authors examined possible links between mtDNA composition and maximum lifespan of multicellular eukaryotes, including 102 mammals. MLS correlates positively with cytosine and negatively with adenine or thymine content, whereas guanine has no apparent effect. This is especially noted for primates. It appears that an increase in MLS of mammals is associated with thymine-to-cytosine substitution. The results suggest that the MLS may be associated with stability and/or mutability of mtDNA and call for further investigation of the mitochondrial genome as a potential target for lifespan extending interventions. PMID- 16706649 TI - Relationship between aging and susceptibility of erythrocytes to oxidative damage: in view of nutraceutical interventions. AB - Twelve (12) healthy elderly subjects were divided into two groups: (a) those given an antioxidant/NO-modulating fermented papaya preparation (FPP) 9 g/day for 4 weeks, and (b) a placebo group. No protein/lipid distribution in erythrocytes (RBC) membranes was noted among different ages and treatments. Higher RBC concentration of malondialdehyde and nitric oxide synthase were found in the elderly (p < 0.05 versus "young" controls), whereas superoxide dismutase was unaltered. Such abnormalities were prevented by FPP supplementation (p < 0.01). RBC and RBC ghosts showed an enhanced susceptibility to lipid peroxidation by using cumene hydroperoxide (p < 0.01 versus young) but FPP supplementation significantly protected intact RBC (p < 0.05). These preliminary data suggest that nutraceuticals with antioxidant/NO-regulating properties significantly protect from RBC oxidative damage, and are potential weapons for the aging process and chronic and degenerative diseases. PMID- 16706650 TI - Aging-related cell surface ECTO-NOX protein, arNOX, a preventive target to reduce atherogenic risk in the elderly. AB - A family of constitutive cell surface ECTO-NOX proteins capable of oxidizing reduced quinones, initially described as NADH oxidases, has offered an opportunity to formulate, for the first time, a complete electron transport chain from the cytosol to oxygen at the cell surface with the ECTO-NOX proteins acting as the terminal oxidases. The ECTO-NOX proteins of the cell surface have been postulated as well to link the accumulation of lesions in mitochondrial DNA to cell surface accumulations of reactive oxygen species as one consequence of their role as a terminal oxidase in a plasma membrane electron transport chain. Of the several ECTO-NOX proteins now known, one is a novel cell surface form (arNOX) associated with lymphocytes, sera, saliva and perspiration of patients of age 50 or older and is capable of directly reducing ferric cytochrome c through the generation of superoxide. Because of their cell surface location, ECTO-NOX proteins capable of superoxide generation in response to aging would serve to propagate the aging cascade both to adjacent cells and to oxidize circulating lipoproteins. The generation of superoxide associated with aging is inhibited by coenzyme Q10. As such, the findings provide a rational basis for the antiaging activity of circulating coenzyme Q10 in the prevention of atherosclerosis and other aging-related oxidative changes in cell membranes and circulating lipoproteins. PMID- 16706651 TI - Emerging strategies for the treatment of hereditary metabolic storage disorders. AB - Metabolic storage disorders are caused by mutations in genes that result in insufficient activity of enzymes required for the catabolism of substances that arise from the turnover of senescent cells in the body. Among the most prevalent of these conditions are Gaucher disease and Fabry disease, which are caused by reduced activity of the housekeeping enzymes glucocerebrosidase and alpha galactosidase A, respectively. Enzyme replacement therapy is extraordinarily effective for patients with Gaucher disease. It is under examination in patients with Fabry disease, and improvement of various clinical aspects in these patients has been documented. The blood-brain barrier prevents systemically administered enzymes from reaching the central nervous system. This limitation is a major impediment for the treatment of patients with enzyme deficiency disorders in whom the brain is involved. Alternatives to enzyme replacement therapy that have been initiated to treat systemic manifestations and brain involvement in patients with metabolic disorders include substrate reduction therapy, active site-specific chaperone therapy, and gene therapy. The present status and anticipated advances in the application of these therapeutic approaches are examined here. PMID- 16706652 TI - Advanced atherosclerotic foam cell formation has features of an acquired lysosomal storage disorder. AB - Atherosclerosis is a disease of large- and medium-sized arteries. Complications from atherosclerosis remain a serious cause of morbidity and mortality in industrialized countries. The disease begins very early in life and effects most people in the West. However, because the progression of the disease is slow, symptoms usually do not occur until after the fifth decade of life. Because atherosclerosis is a ubiquitous occurrence throughout the world, as life expectancy is prolonged most populations will see increasing numbers of deaths from complications of atherosclerosis unless there are dramatic advances in treatment. Because it begins so early in life, current treatment is aimed at slowing or reversing the progression of the disease rather than eliminating the initiating steps. Changes in diet and exercise, cholesterol-lowering drugs, and improvements in surgical treatments have made significant inroads into prolonging life, but much work is still required. To proceed further, a better understanding is needed of the underlying causes of disease progression. In this regard, evidence is mounting that the foam cells of the lesion (a critical cell in atherosclerosis progression) exhibit characteristics of an acquired lysosomal storage disorder. In this review the evidence for this conclusion is reviewed and the ramifications of this conclusion are explored with regard to the understanding of disease progression mechanisms, possible improvements in treatment, and their role in increasing life expectancy. PMID- 16706653 TI - Indirect antioxidant protection against photooxidative processes initiated in retinal pigment epithelial cells by a lipofuscin pigment. AB - Oxidative mechanisms are considered to contribute to the aging changes in retinal pigment epithelial (RPE) cells that underlie the pathogenesis of age-related macular degeneration. An important source of oxidative damage is likely to be the photoreactive pigments that progressively accumulate and constitute the lipofuscin of retinal pigment epithelial cells. Evidence for a link between RPE lipofuscin and cellular dysfunction is also provided by the understanding of disease progression in Stargardt disease. Using a culture model previously used to demonstrate photooxidative damage to retinal pigment epithelial cells that have accumulated the lipofuscin fluorophore A2E, it was shown that the propensity for cell death is increased under conditions that deplete cellular levels of glutathione. Additionally, sulforaphane, a phytochemical and inducer of phase 2 enzymes, protected RPE cells that accumulated A2E and were irradiated at 430 nm. The protection afforded by sulforaphane was paralleled by elevated levels of glutathione and increases in the activities of the phase 2 enzymes NAD(P)H:quinone reductase and glutathione-S-transferases. Moreover, transcriptional induction of NAD(P)H:quinone reductase was indicated by the increases in mRNA determined by real time RT-PCR. There has been considerable interest in the intake of carotenoids and antioxidant vitamins and the related incidence of age-related macular degeneration. The present results indicate that the indirect antioxidant activity of plant-derived phase 2 inducers also may be potentially important. PMID- 16706654 TI - Prevention and repair of protein damage by the Maillard reaction in vivo. AB - The aging human extracellular matrix (ECM) and tissues rich in long-lived proteins undergo extensive changes with age that include increased stiffening, loss of elasticity, insolubilization, and decreased proteolytic digestibility. Most if not all these changes can be duplicated by the Maillard reaction in vitro, that is, the incubation of the proteins with reducing sugars and oxoaldehydes. These carbonyls eventually form advanced glycation end products (AGEs) and crosslinks that impair proteolytic digestibility and alter protein conformation. To date, close to 20 AGEs have been found in the human skin, of which ornithine is the single major result of damage to arginine residues, and glucosepane the single major crosslink. Although redox active metals and oxoaldehydes appear to play an important role in protein damage in experimental diabetes, their role in diabetic humans is still poorly understood. Evidence for the existence of deglycating enzymes has been found in vertebrates, bacteria, and fungi. However, only the vertebrate enzymes can deglycate larger, intracellular proteins via an ATP-dependent mechanism. Protein engineering will thus be needed to adapt Amadoriase enzymes toward deglycation of ECM proteins for purpose of probing the role of advanced glycation in animal models of diabetes and age related diseases. The blocking of the reactivity of the glucosepane precursor using potent nucleophiles may be useful in preventing age-related changes in ECM proteins. However, there currently is no evidence in support of the proposed ability of so-called "AGE breakers" to cleave existing crosslinks of the Maillard reaction in vivo, and other mechanisms of action should be sought for this class of compounds. PMID- 16706655 TI - Extracellular glycation crosslinks: prospects for removal. AB - Extracellular aging--accumulating molecular damage by glycation, oxidation, and crosslinking of long-lived extracellular proteins, mainly collagen and elastin- is a major cause of several important human aging pathologies. Crosslinking increases mechanical stiffness of blood vessels and urinary bladder. Crosslinking impairs the functioning of the kidney, heart, retina, and other tissues and organs. Glycation adducts trigger inflammatory signaling, provoking tissue damage and cancers. Crosslinking tightens up the extracellular matrix (ECM), hardening it against natural turnover processes. Known crosslink breakers (e.g., alagebrium, of the thiazolium halide family) are only partly effective because they break only a subset of AGE crosslink structures (sugar-derived alpha diketone bridges). So far, no agent has been found that breaks the prevalent glucosepane and K2P crosslink structures. Enzymes that would be able to recognize and disassemble glycation products may be too big to migrate into the ECM and repair collagen or elastin in vivo. Two approaches to therapy development are presented here. ECM turnover enhancement would enhance natural processes to digest old ECM and replace it with new. It will be important to tune the collagen degradation to a rate slow enough to prevent dire side-effects, such as hemorrhage from leaky blood vessels as collagen molecules are removed and replaced. Glycation breaker discovery would use high-throughput screening and rational drug design to find molecules that are able to break glucosepane crosslinks and K2P crosslinks of extracellular proteins. Candidates would be further screened for selectivity and toxicity in order to avoid damage to other molecules. PMID- 16706656 TI - Cryopreservation of complex systems: the missing link in the regenerative medicine supply chain. AB - Transplantation can be regarded as one form of "antiaging medicine" that is widely accepted as being effective in extending human life. The current number of organ transplants in the United States is on the order of 20,000 per year, but the need may be closer to 900,000 per year. Cadaveric and living-related donor sources are unlikely to be able to provide all of the transplants required, but the gap between supply and demand can be eliminated in principle by the field of regenerative medicine, including the present field of tissue engineering through which cell, tissue, and even organ replacements are being created in the laboratory. If so, it could allow over 30% of all deaths in the United States to be substantially postponed, raising the probability of living to the age of 80 by a factor of two and the odds of living to 90 by more than a factor of 10. This promise, however, depends on the ability to physically distribute the products of regenerative medicine to patients in need and to produce these products in a way that allows for adequate inventory control and quality assurance. For this purpose, the ability to cryogenically preserve (cryopreserve) cells, tissues, and even whole laboratory-produced organs may be indispensable. Until recently, the cryopreservation of organs has seemed a remote prospect to most observers, but developments over the past few years are rapidly changing the scientific basis for preserving even the most difficult and delicate organs for unlimited periods of time. Animal intestines and ovaries have been frozen, thawed, and shown to function after transplantation, but the preservation of vital organs will most likely require vitrification. With vitrification, all ice formation is prevented and the organ is preserved in the glassy state below the glass transition temperature (T(G)). Vitrification has been successful for many tissues such as veins, arteries, cartilage, and heart valves, and success has even been claimed for whole ovaries. For vital organs, a significant recent milestone for vitrification has been the ability to routinely recover rabbit kidneys after cooling to a mean intrarenal temperature of about -45 degrees C, as verified by life support function after transplantation. This temperature is not low enough for long-term banking, but research continues on preservation below -45 degrees C, and some encouraging preliminary evidence has been obtained indicating that kidneys can support life after vitrification. Full development of tissue engineering and organ generation from stem cells, when combined with the ability to bank these laboratory-produced products, in theory could dramatically increase median life expectancy even in the absence of any improvements in mitigating aging processes on a fundamental level. PMID- 16706657 TI - Cryptobiosis, aging, and cancer: yin-yang balancing of signaling networks. AB - Some organisms adapt to persistent and severe stress by reversibly adjusting life death balance to a new equilibrium, e.g., anhydrobiosis ("life without water") enables survival in a quiescent state on extreme desiccation. Aging is characterized by declining response and increasing vulnerability to stress, and the balance slowly, and irreversibly, tilts toward death. Although tumorigenesis tips the balance of cells to prolonged life, paradoxically it can cause organismal death. At the molecular level, all these phenomena involve complex signaling pathways, but it is highly likely that the overall balance of signaling outcomes, rather than individual signals themselves, plays the pivotal role in life-death decisions. PMID- 16706658 TI - The neurosecretory system is hypertrophied in senescence-accelerated mice. AB - The hypothalamic supraoptic and paraventricular nuclei form the neurosecretory system and synthesize the neurohormones oxytocin and arginine-vasopressin. The senescence-accelerated mouse is a model of rapid aging that displays senile amyloidosis and memory problems. This paper presents the characterization of the neurosecretory system and describes the presence of a bilateral constant cluster of neurosecretory neurons in these mice. The stereologic analysis revealed that these groups contain 197 +/- 18 cells (87% synthesize arginine-vasopressin and 13% oxytocin). The presence of these clusters of neurosecretory neurons suggests that these mice could present greater neurohormone synthesis, increasing the deleterious effects of accelerated aging in this strain. PMID- 16706659 TI - Natural background radioactive carbon and the natural death rate of people. AB - A brief analysis of the known data on the potential danger of radiocarbon incorporation into DNA structure shows that the great genetic importance of transmutational transformations of DNA-incorporated 14C is theoretically and experimentally proved. This effect exists both in huge and small radiation doses (similar to doses of 14C natural background radiation). Therefore, the human death rate can be assumed to be dependent on natural and anthropogenous fluctuations of atmospheric 14C. Calculation methods of the age parameter dynamics of the natural human death rate are offered. It is shown that when calculating the parameters of the natural death rate, the use of Gompertz's formula is reasonable provided that the data on the general death rate are taken for the age interval "60 to 85 years." The ratios reflecting the regular and casual errors of the parameters R and a of Gompertz's equation, caused by people's casual deaths, were determined. A comparison of the historical dynamics of people's natural death rates in the last 150 years with the variations of 14C of the natural background during the same period showed that these are coordinated phenomena, the strong correlation of which indicates the possibility of their functional dependence. The 14C-concentration increase in an organism is the result of its increase in the surrounding biospheric composition, causing an increase in the natural death rate and vice versa: The increase of a person's life expectancy is caused by a decrease of 14C concentration. PMID- 16706660 TI - Radiocarbon mechanism of aging. AB - A hypothetical mechanism of 14C-generated influence on a person's rate of aging is offered. The mathematical description and the following approbation of the radiocarbon mechanism of aging made it possible to coordinate the Gompertz and Strehler-Mildvan laws and predict the existence of one more, till-now unknown, interrelation of the parameters of people's natural death rate. A review of the death rate statistics of the populations of western European countries as well as the United States and Australia for the 19th and 20th centuries gave a positive result. It had been assumed that a person's organism has a "discrimination mechanism" aimed at limiting 14C incorporation into DNA structure and that the efficiency of this mechanism is different for different populations of people. It follows from the results of the research that the activity of such a discrimination function of an organism tends to sharply increase in proportion to an increase of 14C concentration in one's surrounding biosphere. PMID- 16706661 TI - Variations of humans' natural death rate and the radiocarbon aging mechanism. AB - The historical dynamics of the natural death rate have been studied for the populations of 48 European countries from 1970 to 2002, as well as of 191 countries worldwide for 1999, 2000, and 2001. The variations of natural death rate parameters appear to obey the common law in practically all the populations of the planet, and this law follows from the radiocarbon mechanism of aging. The results allow one to imagine the existence of the connection between a person's natural death probability and his or her environmental conditions, brought about by the fact that the environment determines the change of 14C concentration in an organism. Experimental confirmation of such a hypothesis has been considered. As a primary step, it implies the choice of a basic risk factor determining the natural death rate of this or that biologic object, then a comparison of the intensity of the chosen risk factor with radiocarbon content in the whole organism as well as its separate structures. PMID- 16706662 TI - Statistical issues of regression analysis on development of an age-predictive equation. AB - Several statistical information criteria and other statistical issues such as residual examinations and multi-collinearity in regression analysis were argued to establish the predictive equation of age from physical fitness measurements. These independent variables of physical fitness, body composition, two blood pressures, and bone strength were taken from 31 Japanese males (48 +/- 11.9 years old) living in rural areas. The following equation was selected as the optimal one according to Akaike's information criterion: y = 90.0 -1.294 * SITUP - 0.467 * MAP - 0.094 * BALANCE (R = 0.837). When all ten variables were included in the initial equation, R was 0.864, and the highest R* (R adjusted for d.f.) was observed at the fifth step. PMID- 16706663 TI - Soft strategies for postponing aging and prolonging human life. AB - There are some effective soft strategies for postponing aging. One strategy is based on the all-cause mortality studies. It is possible to use many data of epidemiologic studies about different risk factors influencing all-cause mortality. This strategy is carried out to some extent and quite unintentionally. The other strategy is studying and taking into account weak sensations of discomfort to avoid developing intensive symptoms and disorders. The authors have used these strategies for 20 years. Both strategies have the potential to increase the interest of people in science for postponing aging because real results can be seen. PMID- 16706664 TI - Psychosocial aspects and zinc status: is there a relationship with successful aging? AB - It is very interesting and innovative to study the interrelationships between biological characteristics, particularly zinc status, and psychosocial conditions in old age, because there are few and fragmentary data in the literature. The aim of this study was to examine the interrelationship between serum albumin value (an indicator of zinc status) and some psychosocial characteristics in elderly Italian volunteers recruited for the ZINCAGE project, which is supported by the European Commission in the Sixth Framework Programme (Food-CT-2003- 506850). A protocol of tests and questionnaires was used: the Lifestyle Questionnaire, the Mini-Mental Status Examination (MMSE), the Geriatric Depression Scale (GDS, 15 items), and the Perceived Stress Scale. A sample of 174 old subjects were recruited in Region Marche (Central Italy), and classified into three age groups: 65 to 74 years old, 75 to 84 years old, and >85 years old (including some nonagenarians). The preliminary results show that 69.7% of the subjects have no cognitive impairment; 66.5% have a value of the GDS scale indicating no depression; and 17% have an albumin deficiency. The majority of these are >85 years old and women. A relationship between level of albumin (used as indicator of zinc status) and depression has been found: 71% of subjects with albumin deficiency displayed a higher value on the depression test against 29% of subjects with a normal value (p < 0.01). These preliminary results show a relationship between serum albumin and psychological characteristics, in particular depression in an old Italian population. This further suggests that a zinc deficiency, via hypoalbumin values, is involved in impaired psychological characteristics in the elderly. PMID- 16706665 TI - Liver exposure to xenobiotics: the aging factor and potentials for functional foods. AB - Hepatocytes isolated from 20- and 4-month Wistar rats and cultured with or without alpha-linolenic acid (LNA) were then added with nutraceutical YHK or sylibin before the test with iron or copper. Overall, YHK proved to be more effective than sylibin in Fe/Cu-induced peroxidative damage on normal and LNA loaded hepatocytes (p < 0.05). YHK exerted a significant protection against DPPH radical-scavenging activity in the "old" group (p versus sylibin) and against lipophilic generators in both age groups (p < 0.05 versus sylibin). Both compounds were ineffective on age-related increase of surface-charge density. These preliminary data suggest that age per se enhances the vulnerability of hepatocytes to xenobiotics, whereas some safe nutraceuticals seem to exert significant protective effects. PMID- 16706666 TI - The aging gut motility decay: may symbiotics be acting as "implantable" biologic pace-makers? AB - Motility recording of small and large intestine was performed in old Wistar rats divided into three groups: (a) standard diet, (b) standard diet plus a symbiotic preparation, and (c) standard diet plus a heat-inactivated symbiotic preparation. SCM-III. significantly increased the myoelectric activity of small intestine and colon (p < 0.01 versus [a] and [c]) paralleling "young" values of 4-month-old rats and increased the spike burst frequency in the proximal-distal colon (p < 0.05). SCM-III significantly increased the frequency and duration of spike bursts in the jejunum, transverse-distal colon, and defecation frequency, while decreasing the intervals of migrating motor complex in the colon (p < 0.01) to "young" values with an increased mRNA expression of VIP (p < 0.05). Gut flora manipulation aimed to modulate myoelectric activity can tentatively help reversing age-related motility decay. PMID- 16706667 TI - How evolutionary thinking affects people's ideas about aging interventions. AB - Evolutionary theory has guided the development of antiaging interventions in some conscious and some unconscious ways. It is a standard assumption that the body's health has been optimized by natural selection, and that the most benign and promising medical strategies should support the body's efforts to maintain itself. The very concept of natural healing is a reflection of evolutionary thinking about health. Meanwhile, a developing body of experimental evidence points to the startling hypothesis that aging is a metabolic program, under genetic control we are programmed for death. Evolution has provided that the aging program can be abated in times of stress, e.g., caloric restriction. CR mimetics are already recognized as a promising avenue for antiaging research. Beyond this, there are two ancient mechanisms of programmed death in protists that have survived half a billion years of evolution, and still figure in the aging of vertebrates today. These are apoptosis and replicative senescence via telomere truncation. Most researchers have been wary of modifying these mechanisms because they are known to play a stopgap role in cancer prevention. But intriguing evidence suggests that, despite some counter-carcinogenic function, the net result of both these mechanisms may be to shorten lifespan. Thus, interventions that suppress apoptosis and that preserve telomeres may be promising avenues for life extension research. A third element of the body's self destruction program co-opts the inflammation response. Epidemiological evidence suggests that NSAIDs including aspirin protect against atherosclerosis, arthritis, and some forms of cancer. It may be that aging engages an autoimmune response that can be modified by drugs acting more narrowly on this same pathway. The existence of an evolutionary program that controls aging from the top down supports a new optimism concerning the types of antiaging interventions that are possible, and the likelihood that simple strategies may have dramatic results without dramatic side-effects. PMID- 16706668 TI - Zinc homeostasis in aging: two elusive faces of the same "metal". AB - Proteins involved in zinc homeostasis may be altered in aging. This phenomenon may lead to zinc deficiency in the peripheral blood and an accumulation of zinc bound to insoluble aggregates at the extracellular level in the brain. Therefore, it should be more correct to talk about aging as a condition associated with zinc dyshomeostasis rather than deficiency. Restoring functional zinc homeostasis in aging people is an attractive field for antiaging research, but requires further knowledge than the current state of the art. PMID- 16706669 TI - Aging and oral health related to quality of life in geriatric patients. AB - Oral health is more than healthy teeth. Oral diseases and disorders can affect general health, well-being, and quality of life. The goal of this investigation was to establish oral health related to quality of life of geriatric patients. The study was made for a period of 12 months in a private dental office in Sofia, Bulgaria. It included 53 geriatric patients (36 women and 17 men). The women's average age was 69.5 years old (65 to 87), and men's average age was 70.5 years old (65 to 84). A special questionnaire was made and patients were asked about specific dental problems related to quality of life. Of the respondents, 69.8% were women and only 30.2% were men, which means that women are more likely to visit dental health care offices. Of geriatric patients, 31.5% visit the dental office because of acute pain and 68.4% because of caries. The following are the specific items used to assess geriatric oral health and quality of life: trouble biting or chewing (eating), uncomfortable eating in front of other people, trouble speaking, and limitation of social contacts. Oral health can be an indicator of general health and quality of life in geriatric patients. Oral diseases are progressive and cumulative. They become more complex over time. Improved oral health will allow geriatric patients to improve their self confidence, have active social contacts, and restore the ability to work at home or on the job. PMID- 16706670 TI - Is thrombolytic therapy effective in elderly patients? AB - Thrombolytic therapy (TT) is applied in patients (<75 years of age) with acute myocardial infarction (AMI) with ST-segment elevation. It is recommended for elderly patients who have percutaneous transluminal coronary angioplasty (PTCA) or bypass surgery. For PTCA and bypass surgery a cardiosurgery medical team, which can be found in large medical hospitals, has to be prepared. Patients with AMI with ST-segment elevation who are older than 75 and who do not have TT have a dubious prognosis for longevity and quality of life. The aim of this study was to propose recommendations and behavior for reperfusion with TT of the coronary arteries of elderly patients. The investigation was conducted for a period of 5 years (2000 to 2004). Investigators registered 2462 patients, 502 of whom had AMI, and 103 (54 men, 49 women) of whom were older than 75 years. Reteplase (Rapilysin, Hoffman La Roche, Switzerland) was applied to 10 patients. The general and mental state of patients as well as comorbidity were assessed. The H2 blocker Famotidine (Quamatel-Gedeon-Richter, Hungary) was used to prevent bleeding from the gastrointestinal tract. The applied TT had a positive effect on these >75-year-old patients. The oldest woman was 89 years old and the oldest man was 93. There were no complications during the treatment of these elderly patients. Aging related to serious health problems can be improved with the application of TT to elderly patients with AMI, and ST-segment elevation is an excellent reperfusion therapy. It is possible to achieve a dose reduction of the thrombolytic agent, arterial pressure no higher than 160/100 mmHg, individual risk assessment for intracranial hemorrhage, and prevention of bleeding from the gastrointestinal tract. PMID- 16706671 TI - Negligible senescence: how will we know it when we see it? AB - The recent public claim that "SENS is a practical, foreseeable approach to curing aging" has stirred considerable controversy among bio-gerontologists. Testing this hypothesis will not only require precise definitions for the somewhat subjective terms "practical," "foreseeable," and "curing," it will require a precise definition of the term "aging." To facilitate proper experimental design, this definition must focus on the nature of aging itself, not its causes or consequences. Aging in mammals is a process that begins early in adult life and continues steadily thereafter until death. It is manifested by a decline in the functional capacity (or, more precisely, reserve capacity) of a variety of vital physiologic systems leading to increasing risk of morbidity and mortality over time. Aging, however, cannot be measured by simply monitoring morbidity and/or mortality. Aging can only be measured by monitoring the decline of global functional capacity itself. This, in turn, will require an operational definition of aging expressed as a rate function (i.e., it will have units expressing aging as an overall rate of functional change per unit time). Widespread acceptance of such global indexes of aging rate in animal models and humans will greatly facilitate research activity specifically designed to increase the understanding of aging mechanisms and antiaging interventions. PMID- 16706672 TI - Living and all-time world longevity record-holders over the age of 110. PMID- 16706674 TI - Targeting transcription factors in cell regulation. AB - The author compares transcription factors (TFs) with 13 other protein classes from the point of their ability to regulate various cellular processes. The comparison is performed using data from the ResNet 4.0 database containing molecular interactions extracted from scientific literature. The author introduces two quantitative characteristics for evaluating the ability of every protein to regulate a cell process. Using these measures, he evaluates the efficiency of TFs and other protein classes to regulate the biological process pathways. It was found that TFs, on average, are not the best class for regulating an individual cell process. They have lower regulatory specificity, (i.e., single TF tends to regulate many different biological processes). TFs also tend to be placed downstream in the biological process pathways, being a target of the regulatory relation more often than being a regulator. Possible implications of these findings for drug development are discussed. PMID- 16706675 TI - Current targets in irritable bowel syndrome: an interview with Eamonn Quigley. Interview by Emma Quigley. AB - Eamonn M M Quigley, MD, FACG, is Vice-President of the World Gastroenterology Organisation and Secretary of the American College of Gastroenterology. He is also Professor of Medicine and Human Physiology and Head of the Medical School at the National University of Ireland in Cork. Between 1991 and 1998, he served as Chief of Gastroenterology and Hepatology at the University of Nebraska Medical Center, where he was also Professor in the Department of Internal Medicine. Dr Quigley has been awarded a Fogarty International Fellowship by the National Institutes of Health and an Ainsworth Traveling Scholarship from University College Cork. He completed higher medical training in gastroenterology and internal medicine as a Senior Registrar and Lecturer in the Department of Medicine at Hope Hospital and the University of Manchester. His MD thesis was awarded by the National University of Ireland in 1984 and he received his medical education at University College Cork, graduating MB BCh BAO in 1976. Dr Quigley served as Editor-in-Chief of the American Journal of Gastroenterology from 1997 to 2003, and he has published > 400 articles, including original manuscripts, editorials, review articles, book chapters and case reports. He is also interested and involved in education in the area of gastroenterology and has participated in, or directed symposia, workshops and other teaching forums and prepared a variety of related teaching aids. PMID- 16706676 TI - Targets for deep brain stimulation in Parkinson's disease. AB - The use of stimulation electrodes implanted in the brain to control severely disabling neurological and psychiatric conditions is an exciting and fast emerging area of neuroscience. An excellent example is Parkinson's disease (PD), in which tens of thousands of patients have now been implanted with stimulation electrodes. Patients with PD underwent deep brain stimulation (DBS) at the level of the thalamus, globus pallidus internus, subthalamic nucleus, pedunculopontine nucleus and prelemniscal radiation. The results of these interventions revealed that each target has its own specific stimulation-related positive and negative effects. Clinicians can choose their DBS target based on the situation of their individual PD patients. In the authors' opinion, patient-specific targeting should be preferred over disease-specific targeting. In this review, the authors give an overview of the targets that have been used for DBS in PD and discuss patient-specific targeting. PMID- 16706677 TI - Regeneration following spinal cord injury, from experimental models to humans: where are we? AB - Regeneration in the adult CNS following injury is extremely limited. Traumatic spinal cord injury causes a permanent neurological deficit followed by a very limited recovery due to failed regeneration attempts. In fact, it is now clear that the spinal cord intrinsically has the potential to regenerate, but cellular loss and the presence of an inhibitory environment strongly limit tissue regeneration and functional recovery. The molecular mechanisms responsible for failed regeneration are starting to be unveiled. This gain in knowledge led to the design of therapeutic strategies aimed to limit the tissue scar, to enhance the proregeneration versus the inhibitory environment, and to replace tissue loss, including the use of stem cells. They have been very successful in several animal models, although results are still controversial in humans. Nonetheless, novel experimental approaches hold great promise for use in humans. PMID- 16706678 TI - Targeting glycogen synthase kinase-3 as an approach to develop novel mood stabilising medications. AB - Historically, success in the pharmacological treatment of bipolar disorder has arisen either from serendipitous findings or from studies with drugs (antipsychotics and anticonvulsants) developed for other indications (schizophrenia and epilepsy, respectively). Lithium has been in widespread clinical use in the treatment of bipolar disorder for > 30 years. Development of lithium-mimetic compounds has the potential to result in a more specific medication, with fewer side effects and a less narrow dose range. However, novel medications based upon a known mechanism of action of this drug are yet to be developed. Increasing evidence suggests that a next-generation lithium compound may derive from knowledge of a direct target of lithium, glycogen synthase kinase 3 (GSK-3). GSK-3 is an intracellular enzyme implicated as a critical component in many neuronal signalling pathways. However, despite the large body of preclinical data discussed in this review, definitive validation of GSK-3 as therapeutically relevant target of lithium will require clinical trials with novel GSK-3 inhibitors. A number of recent reports suggest that it is possible to develop selective, small-molecule GSK-3 inhibitors. PMID- 16706679 TI - Targeted therapy for short QT syndrome. AB - Identified in 2000, short QT syndrome is an electrical disease of the heart characterised as a channelopathy. At first considered extremely rare, families with this disease have been found in Brazil, Finland, Germany, Spain, the Netherlands, France, Turkey, Italy and the US. The focus of the paper is to present a current review of short QT syndrome, as well as providing an overview upon the potential molecular target-based strategies for management of this very deadly disease. Abnormalities in three different potassium channels have been recognised as the cause of the disease and targets for therapy will be discussed for each potassium channel individually. In addition to pharmacological strategies, gene therapy with transfer of genes coding for specific ion channel subunits or regulatory proteins are discussed. PMID- 16706680 TI - Targeting maspin in endothelial cells to induce cell apoptosis. AB - The diseases of cancer remain as some of the leading causes of death in the industrialised world, although there are a multitude of technologies being used in the field of medical oncology to combat these diseases and scientific research continues to make discoveries to improve patient outcomes. Some of this research has focused on the maspin gene and protein. Maspin is predicted to be a unique serpin with tumour suppressor activity. Recent studies have explored the use of maspin as a therapeutic agent against cancer. In one study, maspin was found to inhibit cancer growth and metastasis in a breast cancer mouse model through a maspin DNA-liposome therapy. A separate study showed the ability of maspin to induce apoptosis in tumour-specific endothelial cells. Taken together, these studies demonstrate the potential use of maspin as a viable anticancer therapeutic agent. PMID- 16706681 TI - FXR: a target for cholestatic syndromes? AB - The nuclear farnesoid X receptor (FXR) plays a pivotal role in maintaining bile acid homeostasis by regulating key genes involved in bile acid synthesis, metabolism and transport, including CYP7A1, UGT2B4, BSEP, MDR3, MRP2, ASBT, I BABP, NTCP and OSTalpha-OSTbeta in humans. Altered expression or malfunction of these genes has been described in patients with cholestatic liver diseases. This review examines the rationale for the use of FXR ligand therapy in various cholestatic liver disorders and includes potential concerns. PMID- 16706682 TI - Targeting the oxytocin receptor to relax the myometrium. AB - Oxytocin acting via its receptor is involved in the myometrial hyperactivity of preterm labour and possibly also in that of primary dysmenorrhoea. The closely related hormone vasopressin acting on its uterine receptor of type V1a may also contribute to the myometrial hyperactivity of these conditions. Several pharmaceutical compounds inhibiting these receptors are, therefore, under development and one substance, atosiban, has now been registered in many countries for the treatment of preterm labour. This compound blocks both the oxytocin and the vasopressin V1a receptor. The efficacy is at least as pronounced as that of other types of drugs and side effects are much reduced. In this overview, present knowledge about receptor-mediated effects of oxytocin and vasopressin on myometrial activity is summarised. Furthermore, the therapeutic use of oxytocin and vasopressin V1a receptor antagonists in preterm labour and primary dysmenorrhoea is discussed. PMID- 16706683 TI - Targeting glycogen synthase kinase-3 in insulin signalling. AB - The renewed interest in an enzyme first discovered over 25 years ago stems from the potential of inhibitors of this enzyme to treat conditions as diverse as diabetes, Alzheimer's disease, stroke and bipolar disorder, and even to enhance the repopulating capacity of transplanted haematopoietic stem cells. The emergence of the first few potent and specific glycogen synthase kinase-3 (GSK-3) inhibitors will end years of speculation on their potential and finally allow the impact of GSK-3 inhibitors to be evaluated clinically. The next few years are likely to be particularly exciting ones for fans of this old enzyme. This review focuses on the role of GSK-3 in the insulin signalling pathway and highlights the evidence implicating the enzyme in insulin resistance. Pharmacological in vitro and in vivo proof-of-concept studies are also discussed, which establish the therapeutic potential of GSK-3 inhibitors as agents for the treatment of Type 2 diabetes. PMID- 16706684 TI - Nitric oxide/redox-based signalling as a therapeutic target for penile disorders. AB - Oxidative and/or nitrosative stress is implicated in the pathogeneses of assorted penile disorders of clinical significance, notably erectile dysfunction, priapism and penile fibrosis. It is becoming increasingly recognised that the generation and activity of reactive oxygen and nitrogen species in the penis influence vascular homeostasis of this organ, with adverse effects exerted at cellular and molecular levels. Furthermore, these elements may interact with molecular signalling pathways operating in the penis, modulating their functional roles. This interaction in particular suggests that by accessing molecular targets associated with oxidative/nitrosative stress in the penis, new pharmacotherapeutic approaches may be developed to promote normal erectile ability and preserve erectile tissue health. This notion pertains to, but also extends beyond, interventions which predictably target components of the nitric oxide-based signal transduction pathway for the on-demand treatment of erectile dysfunction. The next line of pharmaceuticals for disorders of the penis, in general, may well spawn from an integrative understanding of the complex regulatory interactions influenced by, as well as influencing nitric oxide signalling in this organ. PMID- 16706685 TI - STAT3: a potential therapeutic target in dendritic cells for the induction of transplant tolerance. AB - Dendritic cells (DCs) control the segue from innate to adaptive immunity. Moreover, depending upon their milieu, DCs can either induce or inhibit immune responses. Whether DCs are immune stimulatory or tolerogenic apparently rests with whether or not the DCs express activated signal transducer and activator of transcription-3 (STAT3), the transcription factor induced by IL-6-like cytokines and IL-10. DCs expressing activated STAT3 produce less IL-12, which results in less effector T cell development. Moreover, DCs expressing activated STAT3 also express the tryptophan-catabolising enzyme indoleamine 2,3-dioxygenase. The kynurenine products of tryptophan catabolism induce T cell apoptosis; this area is of major interest to researchers working on tolerogenic DCs. In various disease models ranging from tumours to autoimmune diseases, administration of STAT3-activating cytokines resulted in attenuation of immune responses. Other corroborating evidence was obtained using conditional STAT3-deficient mice, or mice defective in cytokine signalling. Thus, persistently activating STAT3 in DCs may be a feasible strategy for controlling allograft rejection. PMID- 16706686 TI - Targeting tumour necrosis factor-alpha in corticosteroid-dependent asthma. AB - Asthma is a chronic inflammatory disease of the airways characterised by bronchospasm attacks triggered by various stimuli. The underlying pathogenic mechanism is eosinophil-dominated inflammation causing bronchial hyper responsiveness, but in more severe asthma subjects, neutrophil cells and their directly related cytokines, such as TNF-alpha, are known to play a role in the inflammatory process and also in diminished corticosteroid sensitivity. The bronchospasm attacks are usually relieved by short-acting beta2-agonists, whereas maintenance therapy is represented mainly by inhaled corticosteroids. However, in severe persistent asthma or in difficult to treat asthma these can be ineffective even at high doses. Therefore, other therapeutic alternatives are needed, and etanercept, a TNF-alpha blocking agent, could represent one of them provided the safety and efficacy data are supportive. PMID- 16706687 TI - The second annual symposium on the future of lung cancer: a translational focus. 11-12 November 2005, Washington D.C., USA. AB - Lung cancer continues to be a leading cause of death in the US, and in its most advanced stages remains incurable. Cytotoxic chemotherapies have been the standard of care for the treatment of unresectable disease. However, recent advances in the development of epidermal growth factor receptor (EGFR) inhibitors have led the way to a new generation of targeted biological agents. During the second annual symposium entitled 'the future of lung cancer: a translational focus', which was sponsored by the Physician s Education Resource, new strategies for the treatment of lung cancer were discussed. Besides the role of EGFR inhibitors, potential targets include the angiogenesis pathway; other growth factor pathways, such as phosphoinositol-3 kinase/Akt and Raf-MEK; the 26S proteasome, the histone deacetylase mechanism; and the TNF-related apoptosis inducing factor receptors. Agents that are directed against these targets are all in varying stages of clinical development. As more is learned about their mechanisms of action and clinical spectrum of activity, the author anticipates their incorporation into novel regimens with enhanced activity against lung cancer. PMID- 16706688 TI - Photobiomodulation of pain in carpal tunnel syndrome: review of seven laser therapy studies. AB - In this review, seven studies using photoradiation to treat carpal tunnel syndrome (CTS) are discussed: two controlled studies that observed real laser to have a better effect than sham laser, to treat CTS; three openprotocol studies that observed real laser to have a beneficial effect to treat CTS; and two studies that did not observe real laser to have a better effect than a control condition, to treat CTS. In the five studies that observed beneficial effect from real laser, higher laser dosages (9 Joules, 12-30 Joules, 32 J/cm(2), 225 J/cm(2)) were used at the primary treatment sites (median nerve at the wrist, or cervical neck area), than dosages in the two studies where real laser was not observed to have a better effect than a control condition (1.8 Joules or 6 J/cm(2)). The average success rate across the first five studies was 84% (SD, 8.9; total hands = 171). The average pain duration prior to successful photoradiation was 2 years. Photoradiation is a promising new, conservative treatment for mild/moderate CTS cases (motor latency < 7 msec; needle EMG, normal). It is cost-effective compared to current treatments. PMID- 16706689 TI - Photoengineering of tissue repair in skeletal and cardiac muscles. AB - This review discusses the application of He-Ne laser irradiation to injured muscles at optimal power densities and optimal timing, which was found to significantly enhance (twofold) muscle regeneration in rats and, even more, in the cold-blooded toads. Multiple and frequent (daily) application of the laser in the toad model was found to be less effective than irradiation on alternate days. It was found that in the ischemia/reperfusion type of injury in the skeletal leg muscles (3 h of ischemia), infrared Ga-Al-As laser irradiation reduced muscle degeneration, increased the cytoprotective heat shock proteins (HSP-70i) content, and produced a twofold increase in total antioxidants. In vitro studies on myogenic satellite cells (SC) revealed that phototherapy restored their proliferation. Phototherapy induced mitogen-activated protein kinase/extracellular signal-regulated protein kinase (MAPK/ERK) phosphorylation in these cells, probably by specific receptor phosphorylation. Cell cycle entry and the accumulation of satellite cells around isolated single myofibers cultured in vitro was also stimulated by phototherapy. Phototherapy also had beneficial effects on mouse, rat, dog and pig ischemic heart models. In these models, it was found that phototherapy markedly and significantly reduced (50-70%) the scar tissue formed after induction of myocardial infarction (MI). The phototherapeutic effect was associated with reduction of ventricular dilatation, preservation of mitochondria and elevation of HSP- 70i and ATP in the infarcted zone. It is concluded that phototherapy using the correct parameters and timing has a markedly beneficial effect on repair processes after injury or ischemia in skeletal and heart muscles. This phenomenon may have clinical applications. PMID- 16706690 TI - Clinical and experimental applications of NIR-LED photobiomodulation. AB - This review presents current research on the use of far-red to near-infrared (NIR) light treatment in various in vitro and in vivo models. Low-intensity light therapy, commonly referred to as "photobiomodulation," uses light in the far-red to near-infrared region of the spectrum (630-1000 nm) and modulates numerous cellular functions. Positive effects of NIR-light-emitting diode (LED) light treatment include acceleration of wound healing, improved recovery from ischemic injury of the heart, and attenuated degeneration of injured optic nerves by improving mitochondrial energy metabolism and production. Various in vitro and in vivo models of mitochondrial dysfunction were treated with a variety of wavelengths of NIR-LED light. These studies were performed to determine the effect of NIR-LED light treatment on physiologic and pathologic processes. NIRLED light treatment stimulates the photoacceptor cytochrome c oxidase, resulting in increased energy metabolism and production. NIR-LED light treatment accelerates wound healing in ischemic rat and murine diabetic wound healing models, attenuates the retinotoxic effects of methanol-derived formic acid in rat models, and attenuates the developmental toxicity of dioxin in chicken embryos. Furthermore, NIR-LED light treatment prevents the development of oral mucositis in pediatric bone marrow transplant patients. The experimental results demonstrate that NIR-LED light treatment stimulates mitochondrial oxidative metabolism in vitro, and accelerates cell and tissue repair in vivo. NIR-LED light represents a novel, noninvasive, therapeutic intervention for the treatment of numerous diseases linked to mitochondrial dysfunction. PMID- 16706691 TI - Pro- and anti-inflammatory cytokine content in human peripheral blood after its transcutaneous (in vivo) and direct (in vitro) irradiation with polychromatic visible and infrared light. AB - OBJECTIVE: The aim of this randomized, placebo-controlled, double-blind trial was to investigate changes in the content of 10 cytokines in the human peripheral blood after transcutaneous and in vitro irradiation with polychromatic visible and infrared (IR) polarized light at therapeutic dose. BACKGROUND DATA: The role of cytokines in development of anti-inflammatory, immunomodulatory, and wound healing effects of visible and IR light remains poorly studied. METHODS: The sacral area of volunteers was exposed (480-3400 nm, 95% polarization, 12 J/cm(2)); in parallel, the blood samples of the same subjects were irradiated in vitro (2.4 J/cm(2)). Determination of cytokine content was performed using enzyme linked immunosorbent assay (ELISA). RESULTS: A dramatic decrease in the level of pro-inflammatory cytokines TNF-alpha, IL-6, and IFN-gamma was revealed: at 0.5 h after exposure of volunteers (with the initial parameters exceeding the norm), the cytokine contents fell, on average, 34, 12, and 1.5 times. The reduced concentrations of TNF-alpha and IL-6 were preserved after four daily exposures, whereas levels of IFN-gamma and IL-12 decreased five and 15 times. At 0.5 h and at later times, the amount of anti-inflammatory cytokines was found to rise: that of IL-10 rose 2.7-3.5 times (in subjects with normal initial parameters) and of TGF-beta1 1.4-1.5 times (in the cases of its decreased level). A peculiarity of the light effect was a fast rise of IFN-gamma at 3.3-4.0 times in subjects with normal initial values. The content of IL-1beta, IL-2, IFN-alpha, and IL-4 did not change. Similar regularities of the light effects were recorded after in vitro irradiation of blood, as well as on mixing the irradiated and non-irradiated autologous blood at a volume ratio 1:10 (i.e., at modeling the events in a vascular bed of the exposed person when a small amount of the transcutaneously photomodified blood contacts its main circulating volume). CONCLUSION: Exposure of a small area of the human body to light leads to a fast decrease in the elevated pro-inflammatory cytokine plasma content and to an increase in the the anti-inflammatory factor concentration, which may be an important mechanism of the anti-inflammatory effect of phototherapy. These changes result from transcutaneous photomodification of a small volume of blood and a fast transfer of the light-induced changes to the entire pool of circulating blood. PMID- 16706692 TI - Photobiomodulation of pain and inflammation in microcrystalline arthropathies: experimental and clinical results. AB - OBJECTIVE: This article presents the results of laser therapy in crystal (hydroxyapatite, calcium pyrophosphate, and urates) deposition-induced arthritis in rats and the clinical applications in humans. BACKGROUND DATA: Microcrystalline arthropathies are prevalent among geriatric patients, who are more vulnerable to the side effects of drugs. The effectiveness of laser therapy for pain relief, free of side effects, has been reported in painful conditions. METHODS: Two milligrams of each of the above-mentioned crystals was injected in both joints of the back limbs in three groups of rats; these groups were then treated with laser irradiation. Three other groups received no treatment after the injections. We determined the plasmatic levels of inflammatory markers (fibrinogen, prostaglandin E2, and TNF(alpha)), tissues (prostaglandin E(2)) and conducted anatomopathological studies. Twenty-five patients with acute gout arthritis were randomized into two groups and treated over 5 days: group A, diclofenac 75 mg orally, twice a day; and group B, laser irradiation once a day. Forty-nine patients with knee chronic pyrophosphate arthropathy were randomized into two groups and treated over 21 days; group A, diclofenac 50 mg orally, twice a day; and group B, laser irradiation once a day. Thirty patients with shoulder chronic hydroxyapatite arthropathy were randomized into two groups and treated over 21 days; group A, diclofenac 50 mg orally, twice a day; and group B, laser irradiation once a day. RESULTS: Fibrinogen, prostaglandin E(2), and TNF(alpha) concentrations in the rats injected with crystals and treated with laser decreased significantly as compared with the groups injected with crystals without treatment. Both laser therapy and diclofenac achieved rapid pain relief in patients with acute gouty arthritis without significant differences in efficacy. Laser therapy was more effective than diclofenac in patients with chronic pyrophosphate arthropathy and in patients with chronic apatite deposition disease. CONCLUSION: Laser therapy represents an effective treatment in the therapeutic arsenal of microcrystalline arthropathies. PMID- 16706693 TI - Photoengineering of neural tissue repair processes in peripheral nerves and the spinal cord: research development with clinical applications. AB - The purpose of the present paper is to provide our data on the effects of phototherapy on peripheral nerve recovery. The aim is to call attention to an issue that still requires much research to elucidate the biological mechanisms through which phototherapy exerts its effects on nerve tissue, and to provide clinicians with the basis for planning clinical trials on the use of phototherapy for enhancing post-traumatic nerve regeneration. PMID- 16706694 TI - Low-level laser therapy in acute pain: a systematic review of possible mechanisms of action and clinical effects in randomized placebo-controlled trials. AB - OBJECTIVE: The aim of this study was to review the biological and clinical short term effects of photoradiation in acute pain from soft-tissue injury. BACKGROUND DATA: It is unclear if and how photoradiation can reduce acute pain. METHODS: Literature search of (i) controlled laboratory trials investigating potential biological mechanisms for pain relief and (ii) randomized placebo-controlled clinical trials which measure outcomes within the first 7 days after acute soft tissue injury. RESULTS: There is strong evidence from 19 out of 22 controlled laboratory studies that photoradiation can modulate inflammatory pain by reducing levels of biochemical markers (PGE(2), mRNA Cox 2, IL-1beta, TNFalpha), neutrophil cell influx, oxidative stress, and formation of edema and hemorrhage in a dose-dependent manner (median dose 7.5 J/cm(2), range 0.3-19 J/cm(2)). Four comparisons with non-steroidal anti-inflammatory drugs (NSAIDs) in animal studies found optimal doses of photoradiation and NSAIDs to be equally effective. Seven randomized placebo-controlled trials found no significant results after irradiating only a single point on the skin overlying the site of injury, or after using a total energy dose below 5 Joules. Nine randomized placebo controlled trials (n = 609) were of acceptable methodological quality, and irradiated three or more points and/or more than 2.5 cm(2) at site of injury or surgical incision, with a total energy of 5.0-19.5 Joules. Results in these nine trials were significantly in favor of photoradiation groups over placebo groups in 15 out of 18 outcome comparisons. Poor and heterogeneous data presentation hampered statistical pooling of continuous data. Categorical data of subjective improvement were homogeneous (Q-value = 7.1) and could be calculated from four trials (n = 379) giving a significant relative risk for improvement of 2.7 (95% confidence interval [CI], 1.8-3.9) in a fixed effects model. CONCLUSION: photoradiation can modulate inflammatory processes in a dose-dependent manner and can be titrated to significantly reduce acute inflammatory pain in clinical settings. Further clinical trials with adequate photoradiation doses are needed to precisely estimate the effect size for photoradiation in acute pain. PMID- 16706695 TI - Photoengineering of bone repair processes. AB - OBJECTIVE: This paper aims to report the state of the art with respect to photoengineering of bone repair using laser therapy. BACKGROUND DATA: Laser therapy has been reported as an important tool to positively stimulate bone both in vivo and in vitro. These results indicate that photophysical and photochemical properties of some wavelengths are primarily responsible for the tissue responses. The use of correct and appropriate parameters has been shown to be effective in the promotion of a positive biomodulative effect in healing bone. METHODS: A series of papers reporting the effects of laser therapy on bone cells and tissue are presented, and new and promising protocols developed by our group are presented. RESULTS: The results of our studies and others indicate that bone irradiated mostly with infrared (IR) wavelengths shows increased osteoblastic proliferation, collagen deposition, and bone neorformation when compared to nonirradiated bone. Further, the effect of laser therapy is more effective if the treatment is carried out at early stages when high cellular proliferation occurs. Vascular responses to laser therapy were also suggested as one of the possible mechanisms responsible for the positive clinical results observed following laser therapy. It still remains uncertain if bone stimulation by laser light is a general effect or if the isolate stimulation of osteoblasts is possible. CONCLUSION: It is possible that the laser therapy effect on bone regeneration depends not only on the total dose of irradiation, but also on the irradiation time and the irradiation mode. The threshold parameter energy density and intensity are biologically independent of one another. This independence accounts for the success and the failure of laser therapy achieved at low-energy density levels. PMID- 16706696 TI - Photochemistry and photobiology of light absorption by living cells. AB - In this review, we summarize a part of our research concerning photobiostimulative effects on cardiomyocytes, sperm cells, and nerve cells. We concentrate on results demonstrating that photobiostimulation can be described by the Arndt-Schultz (A.S.) curve. Results monitoring an increase in reactive oxygen species (ROS) concentration following visible light irradiation describe the ascending part of the A.S. curve, whereas those that describe the antioxidant role of photobiostimulation represent the descending part of the curve. PMID- 16706698 TI - Photoradiation and orthodontic movement: experimental study with canines. AB - OBJECTIVE: The aim of this study was to evaluate, through a double-blind study, the effect of gallinium-aluminium- arsenic (GaAlAs) laser irradiation on the speed of orthodontic movement in canine premolars. METHODS: Eighteen dogs were divided into two groups, and their third molars were extracted. An orthodontic device was placed between the first molar and the second premolar for stabilization purpose. Group I was irradiated with a dosage of 5.25 J/cm(2) on the right side, whereas the left side was used as the control group. Group II was submitted to the same procedure, but was irradiated with a dosage of 35.0 J/cm(2). Irradiations were done every 7 days, for a total of nine irradiations. The orthodontic space was measured every 21 days. RESULTS: The 5.25 J/cm(2) dosage accelerated orthodontic movement during the first observation period, from 0 to 21 days (p < 0.05), whereas the 35.0 J/cm(2) dosage retarded the orthodontic movement in the treated group when compared with the control group, during both the first and second observation periods, from 0 to 42 days (p < 0.05). CONCLUSION: The results suggets that photoradiation may accelerate orthodontic movement at a dosage of 5.25 J/cm(2), whereas a higher dosage, 35.0 J/cm(2), may retard it. PMID- 16706697 TI - Griseofulvin enhances the effect of aminolevulinic acid-based photodynamic therapy in vitro. AB - OBJECTIVE: In this study, we investigated whether or not griseofulvin (GF), which is an antimycotic widely used for the oral treatment of skin fungal infections, enhanced the effect of aminolevulinic acid-based photodynamic therapy (ALA-PDT) in vitro, using several tumor cell lines. METHODS: A human squamous cell carcinoma line (KB), two human osteosarcoma cell lines from mandible (HOSM-1, HOSM-2), and the human gingiva-derived fibroblast line (HF), representing normal cells, were used. GF enhancement of ALA-PDT was evaluated by comparing the effect of ALAin combination with GF to the effect of ALAalone (GF enhancement rate of ALA-PDT). Also, the effect of GF on intracellular accumulation of protoporphyrin IX (PpIX) was evaluated by comparing the intracellular accumulation of PpIX in the ALA and GF combined treatment with that of ALA treatment alone (pGF enhancement rate of intracellular PpIX). RESULTS: GFenhancement rate of ALA-PDT was 2.51 in KB cells, and 1.65 and 1.27 in HOSM-1 and HOSM-2 cells, respectively. GF enhancement rates of intracellular PpIX were 1.94 in KB cells, 1.53 in HOSM-1 cells, and 1.19 in HOSM-2 cells. GF enhancement rate of intracellular PpIX followed the same trends as the levels of GF enhancement rate of ALA-PDT in the different cell types. For HF, a large effect was not revealed in this study. CONCLUSION: The present study, although preliminary, strongly suggests that concomitant treatment with ALAand GF may be very useful to enhance the effect of ALA-PDT. PMID- 16706699 TI - Steroid receptor antagonist mifepristone inhibits the anti-inflammatory effects of photoradiation. AB - OBJECTIVE: We designed an animal pleurisy study to assess if the anti inflammatory effect of photoradiation could be affected by concomitant use of the cortisol antagonist mifepristone. BACKGROUND DATA: Although interactions between photoradiation and pharmacological agents are largely unknown, parallel use of steroids and photoradiation is common in the treatment of inflammatory disorders such as arthritis and tendinitis. METHODS: Forty BALB/c male mice were randomly divided in five groups. Inflammation was induced by carrageenan administered by intrathoracic injections. Four groups received carrageenan, and one control group received injections of sterile saline solution. At 1, 2, and 3 h after injections, photoradiation irradiation was performed with a dose of 7.5 J/cm(2). Two of the carrageenan-injected groups were pre-treated with orally administered mifepristone. RESULTS: Total leukocyte cell counts revealed that in carrageenan induced pleurisy, photoradiation significantly reduced the number of leukocyte cells (p < 0.0001, mean 34.5 [95% CI: 32.8-36.2] versus 87.7 [95% CI: 81.0 94.4]), and that the effect of photoradiation could be totally blocked by adding the cortisol antagonist mifepristone (p < 0.0001, mean 34.5 [95% CI: 32.1-36.9] versus 82.9 [95%CI: 70.5-95.3]). CONCLUSION: The steroid receptor antagonist mifepristone significantly inhibited the anti-inflammatory effect of photoradiation. Commonly used glucocorticoids are also known to down-regulate steroid receptors, and further clinical studies are necessary to elucidate how this interaction may decrease the effect size of photoradiation over time. For this reason, we also suggest that, until further clinical data can be provided, clinical photoradiation trials should exclude patients who have received steroid therapy within 6 months before recruitment. PMID- 16706700 TI - Effects of 830-nm laser, used in two doses, on biomechanical properties of osteopenic rat femora. AB - OBJECTIVE: The aim of this study was to investigate the effects of photoradiation -infrared at 830 nm--used in two doses, on femora of osteopenic rats. BACKGROUND DATA: Osteoporosis has recently been recognized as a major public health problem. Based on stimulatory effects of photoradiation on the proliferation of bone cells, we hypothesized that photoradiation would be efficient in improving bone mass in osteopenic rats. METHODS: Sixty female animals, divided into six groups, were used: sham-operated control (SC), osteopenic control (OC), sham-operated irradiated with the dose of 120 J/cm(2) (I120), osteopenic irradiated with the dose of 120 J/cm(2) (O120), sham-operated irradiated with the dose of 60 J/cm(2) (I60), and osteopenic irradiated with the dose of 60 J/cm(2) (O60) Animals were 90 days old when operated. Laser irradiation was initiated 8 weeks after operation, and it was performed 3 times a week for 2 months. Femora were submitted to a biomechanical test and to a physical properties evaluation. RESULTS: Maximal load of O120 did not show any difference when compared with SC and I120, but it was higher than the O60 group. Wet weight, dry weight, and bone volume of O60 and O120 did not show any difference when compared with SC. CONCLUSION: The results of the present study indicate that photoradiation had stimulatory effects on femora of osteopenic rats, mainly at the dose of 120 J/cm.(2) However, further studies are needed to investigate the effects of different parameters, wavelengths, and sessions of applications on ovariectomized rats. PMID- 16706701 TI - Treatment of chemotherapy-induced oral mucositis with light-emitting diode. AB - OBJECTIVE: The aim of this study was to assess the clinical effectiveness of phototherapy with noncoherent light in the alleviation of chemotherapy-induced mucositis in patients with metastatic cancer. BACKGROUND DATA: Mucositis occurs in more than 40% of chemotherapy-treated patients, significantly reducing the quality of their lives. Many different interventions have been evaluated to reduce oral mucositis. Recently, good results have been achieved by phototherapy with photoradiation, a technique which has virtually no side effects. Some clinical results seem to indicate that also phototherapy through noncoherent light emissions which can be produced by less expensive light sources such as light-emitting diodes (LEDs) may be effective. However, until now, no studies have been available on this subject. METHODS: Twelve patients, aged from 34 to 82, selected on the basis of a diagnosis of chemotherapy-induced oral mucositis, were treated intra-orally through a noncoherent LED emission, wavelength 645 +/- 15 nm, 7.8 mW, fluence 0.99 J/cm(2), three times a day for 1 week. Mucositis was scored daily using the Daily Mucositis Index (DMI), a scale that evaluates the disease evolution through 16 different items. The primary end-point assessed was the time to recovery, from the start of LED treatment, compared to a nonrandomized control group of 12 patients with comparable stomatitis. RESULTS: The median healing time, expressed as the DMI decrease, was 1.7 (range 1-2.8) and, in seven LED-treated patients, was shorter than in the control group. The healing rate (measured as the ratio of the DMIs) increased from 117% to 164%. CONCLUSION: This pilot study shows that LED treatment is safe and capable of reducing the duration of chemotherapy-induced mucositis. This result needs to be confirmed in an adequate phase III study. PMID- 16706704 TI - Bibliography. Laser literature watch. PMID- 16706702 TI - Influence of Er:YAG laser on cavity preparation and surface treatment in microleakage of composite resin restorations. AB - OBJECTIVE: The purpose of this study was to assess microleakage on class V cavities prepared by Er:YAG laser with varying surface treatment. BACKGROUND DATA: There has been little reported research on microleakage at cavities prepared and treated with Er:YAG laser. METHODS: Fifty preparations using Er:YAG laser or a highspeed handpiece were developed. The surface treatment was performed as follows: Er:YAG laser (group 1); 35% phosphoric acid (group 2); laser + acid (group 3); finishing with low-speed + laser + acid (group 4); conventional preparation developed with a high-speed handpiece + acid (group 5- control). The samples were restored with Single Bond/Z250, thermocycled, isolated, and immersed in a 50% AgNO(3) solution. The restorations were sectioned, and the microleakage was measured in milimeters using a specific computer software. Data were analyzed employing ANOVAand Tukey test. RESULTS: Statistical analysis showed that group 5 (G5) obtained the lowest results of microleakage (22.05%), and it was statistically similar (p > 0.05) to G1 (37.7%) and different (p < 0.05) from the other groups (G2 = 50.4%; G3 = 43%; G4 = 44.2%). The occlusal margins (22.62%) demonstrated less microleakage (p < 0.05) than the cervical margins (56.32%). CONCLUSION: The use of Er:YAG laser for cavity preparation and surface treatment negatively influenced the marginal sealing of composite resin restorations. PMID- 16706705 TI - Approaching the CDC's guidelines on the HIV testing of inpatients: physician referral versus nonreferral-based testing. AB - Despite ongoing evidence that one quarter of HIV-infected people in the United States are unaware of their infection, widespread implementation of the Centers for Disease Control and Prevention's 1993 recommendations regarding routine inpatient HIV testing has not occurred. This study compares two HIV testing strategies: the initial phase of inpatient HIV testing (1999-2001) utilized a physician-referral-based system. The second phase (2001-2003) included the first 2 years' experience with having trained HIV counselors directly approach inpatients regarding their willingness to undergo voluntary HIV counseling and testing (VCT) without physician referral. This latter phase was prompted by a patient attitude survey demonstrating favorable responses to unsolicited approaches by staff regarding HIV testing. Barriers to implementing the latter strategy are discussed and initial experience with rapid HIV testing on this service is also presented. Referral-based testing yielded 2.3 patient referrals (6.4% of total admissions) resulting in 1.2 HIV tests and 0.7 counseling only sessions per day. Nonreferral based testing resulted 6.2 HIV tests and another 3.0 counseling-only sessions per day. HIV VCT on an inpatient service is feasible but challenging. Most patients respond favorably to being approached for VCT. Routinely offering HIV tests to inpatients yields higher testing rates than physician referral-based systems and increases the number of patients who know their HIV status. Recommendations for implementing routing HIV testing on an inpatient service are made. PMID- 16706706 TI - Provision of general and HIV-specific health maintenance in middle aged and older patients in an urban HIV clinic. AB - Given the increased prevalence of HIV infection in older individuals, we evaluated the adequacy of HIV-specific health maintenance, age-appropriate cancer screening, and diabetes management in an urban HIV clinic. We randomly selected 222 HIV-positive patients 40 years or older followed at the Johns Hopkins University Moore Clinic between 1999 and 2002. Demographic, clinical, and pharmaceutical data were abstracted from clinic charts. Outcomes of interest were vaccinations, annual rapid plasmin reagin (RPR) testing, and Papanicoloau smears and mammography in women. Logistic regression analyses were performed to identify variables significantly associated with being up to date on vaccinations. The sample was 56% female and 82% African American with a mean age of 50.9 years. Sixty-five percent used tobacco, 10% used alcohol, and 13% used illicit drugs daily. At the time of evaluation, 87% had received the pneumococcal vaccine. Of nonimmune patients, 66% were vaccinated for hepatitis B and 28% for hepatitis A. Eighty-two percent of women were referred for Papanicoloau smears and 56% for mammography. Only 59% completed the Papanicoloau smear, and 31% had mammography. Forty-two percent of patients with diabetes underwent quarterly foot examinations, and 33% had microalbuminuria screening. Risk factors for missed vaccinations include prior AIDS diagnosis (odds ratio [OR] 1.82, 95% confidence interval [CI] 1.55, 3.13), CD4+ cell count less than 50 cells/mm(3) at the time of visit (OR 6.31, 95% CI 1.74, 22.9), and a history of chronic obstructive pulmonary disease (COPD) or asthma (OR 2.54, 95% CI 1.03, 6.28). In summary, HIV positive patients are more likely to receive HIV-specific primary care interventions, especially vaccinations that can be given in clinic, than routine health maintenance screening that required referral and evaluation elsewhere. This suggests that if health maintenance screening can be delivered in the same clinic, usage rates are likely to increase. PMID- 16706707 TI - Becoming a "treatment success": what helps and what hinders patients from achieving and sustaining undetectable viral loads. AB - Highly active antiretroviral therapy (HAART) adherence research has focused predominantly on individuals with less than optimal clinical outcomes; therefore, little is known about the experiences of individuals who sustain undetectable viral loads. The present study used a qualitative method to explore how individuals who have sustained undetectable viral loads account for their success, and to identify challenges, as well as possible needs, for continued success. Participants were 20 patients at an outpatient infectious disease clinic in an urban center. Participants completed two 60-minute interviews. The Critical Incident Technique was used to identify and classify critical incidents linked with sustaining treatment success. Of the 438 critical incidents collected, 316 were identified as helpful and 122 were identified as unhelpful. Helpful categories included resolving ambivalence, using personal strengths, and fostering helpful relationships. Unhelpful categories were mood, lack of social support, financial difficulties, and medication factors. Doing well on antiretroviral therapy is a dynamic process that requires ongoing attention from both the patient and care provider. The results of this study highlight the efforts of patients to maintain their health and remind care providers not to assume that patients are not facing continuous challenges. Findings from the present study suggest that psychosocial factors do contribute to improved clinical outcomes in patients taking HAART. PMID- 16706708 TI - To take or not to take: decision-making about antiretroviral treatment in people living with HIV/AIDS. AB - Knowledge is limited regarding decision-making about antiretroviral treatment (ART) from the patient's perspective. This substudy of a longitudinal study of psychobiologic aspects of long-term survival, conducted in 2003, compares the rationales of HIV-positive individuals (n = 79) deciding to take or not to take ART. Inclusion criteria were HIV/AIDS symptoms, or CD4 nadir less than 350, or viral load greater than 55,000. Those not meeting any criteria for receiving ART (2/2003 U.S. DHHS treatment guidelines) were excluded. Diagnosis was on average 11 years ago; 36% were female, 42% African American, 28% Latino, 24% white, and 6% other. Qualitative content analysis of semistructured interviews identified 10 criteria for the decision to take or not to take ART: CD4/viral load counts (87%), quality of life (85%), knowledge/ beliefs about resistance (66%), mind body beliefs (65%), adverse effects of ART (59%), easy-to-take regimen (58%), spirituality/worldview (58%), drug resistance (41%), experience of HIV/AIDS symptoms (39%), and preference for complementary/alternative medicine (17%). Participants choosing not to take ART (27%) preferred complementary/alternative medicine (r = 0.43, p = 0.001)1, perceived a better quality of life without ART (r = 0.32, p < 0.004), and weighted avoidance of adverse effects of ART more heavily (r = 0.24, p < 0.030) than participants taking ART (73%). Demographic characteristics related to taking ART were having a partner (r = 0.31, p < 0.008) and having health insurance (r = 0.26, p< 0.040). Decisions to take or not to take ART depend not only on patient medical characteristics, but also on individual beliefs about ART, complementary/alternative medicine, spirituality, and mind-body connection. HIV-positive individuals declining treatment place more weight on alternative medicine, avoiding adverse effects and perceiving a better quality of life through not taking ART. PMID- 16706709 TI - A comparison of HIV stigma and disclosure patterns between older and younger adults living with HIV/AIDS. AB - The purpose of this study was to examine the relationships between age, HIV related stigma, and patterns of disclosure. Previous literature has suggested that older age is associated with increased HIV stigma and less disclosure of HIV status. Eighty-eight individuals, 44 between the ages of 20-39 and 44 aged 50 and over were recruited for the study through an AIDS service organization in the Pacific Northwest. Subjects in each group were matched as closely as possible by gender, ethnicity, HIV exposure and diagnosis. In a comparison of sociodemographic characteristics, older adults (50+) were significantly more likely to live alone, and to be retired. Younger adults were significantly more likely to be never married/ partnered, unemployed and be recipients of Medicaid. Bivariate analysis revealed no significant differences in overall stigma scores between groups; however, younger adults were more likely to fear losing their job because of HIV. Older adults were less likely to disclose HIV to relatives, partners, mental health workers, neighbors, and church members than those 20-39 years of age. Pearson product moment correlations found disclosure to be significantly associated with time since diagnosis, heterosexual exposure, ethnicity, use of HIV services, and having a confidant. Stigma was associated with ethnicity, having a confidant, and instrumental social support. In a multiple regressions analysis, 48.4% of the variance in disclosure accounted for by time since first diagnosis, service use, and having a confidant. Service use was the only independent variable significantly associated with stigma, accounting for 21.6% of the variance. PMID- 16706710 TI - Social stigma concerns and HIV medication adherence. AB - The threat of social stigma may prevent people living with HIV from revealing their status to others and serve as a barrier to HIV treatment adherence. We evaluated the effect of such concerns on self-reported treatment adherence using a short, three-item measure among 204 people living with HIV. Overall, the mean age of participants was 40.1 years, 45% were African American, and 80% were male. People with high HIV stigma concerns were 2.5 times less likely to define and interpret the meaning of CD4 count correctly and 3.3 times more likely to be nonadherent to their medication regimen than those with low concerns. Concern over revealing HIV status was the only statistically significant, independent predictor of adherence in multivariate analysis. Clinical care directed to individuals living with HIV should therefore include considerations for patient sensitivity to social stigma, such as modifications to medication schedules and referrals for counseling prior to enrollment in antiretroviral therapies. PMID- 16706711 TI - Demographic and clinical characteristics of HIV-infected inpatients and outpatients at a Cambodian hospital. AB - A retrospective cross-sectional survey of 299 HIV-infected inpatients and outpatients was conducted between March 1999 and June 2000 in Phnom Penh, Cambodia, to define patient demographics and risk factors for HIV-1 infection, and to compare whether symptoms and opportunistic infections (OIs) differ by gender and site of patient care. The population represented one third of HIV infected patients regularly receiving care at the Sihanouk Hospital Center of HOPE. Over one quarter (26%) of the men were soldiers and 27% were farmers or laborers. Eighty-nine percent of the men had visited sex workers, 29% of men and women had unsafe injections in the past, and 18% of women reported a spouse who was HIV-positive. Eighty percent of patients presented with weight loss more than 10% from baseline. Seventy-two percent of patients were diagnosed with two or more concurrent OIs or conditions. Oral candidiasis (p = <0.001), abdominal lymphadenitis (p = 0.03), and two or more concurrent OIs (p = <0.001) were diagnosed more often among men than women. Multivariate logistic regression shows that patients who presented with weight loss more than 10% from baseline are more likely to have one or more OIs or conditions. The results of this survey suggest that the primary risk factor for HIV-infected men presenting to this care facility was visiting sex workers. The pattern of OIs and other HIV-associated conditions indicated that the majority of patients delayed seeking care at the hospital. HIV physicians in Cambodia should be aware of the likelihood for multiple OIs or conditions when patients present weight loss more than 10% from baseline. PMID- 16706712 TI - PEP must begin early. PMID- 16706713 TI - Medicare problems in Washington. PMID- 16706714 TI - Multiple testing methods for ChIP-Chip high density oligonucleotide array data. AB - Cawley et al. (2004) have recently mapped the locations of binding sites for three transcription factors along human chromosomes 21 and 22 using ChIP-Chip experiments. ChIP-Chip experiments are a new approach to the genomewide identification of transcription factor binding sites and consist of chromatin (Ch) immunoprecipitation (IP) of transcription factor-bound genomic DNA followed by high density oligonucleotide hybridization (Chip) of the IP-enriched DNA. We investigate the ChIP-Chip data structure and propose methods for inferring the location of transcription factor binding sites from these data. The proposed methods involve testing for each probe whether it is part of a bound sequence using a scan statistic that takes into account the spatial structure of the data. Different multiple testing procedures are considered for controlling the familywise error rate and false discovery rate. A nested-Bonferroni adjustment, which is more powerful than the traditional Bonferroni adjustment when the test statistics are dependent, is discussed. Simulation studies show that taking into account the spatial structure of the data substantially improves the sensitivity of the multiple testing procedures. Application of the proposed methods to ChIP Chip data for transcription factor p53 identified many potential target binding regions along human chromosomes 21 and 22. Among these identified regions, 18% fall within a 3 kb vicinity of the 5'UTR of a known gene or CpG island and 31% fall between the codon start site and the codon end site of a known gene but not inside an exon. More than half of these potential target sequences contain the p53 consensus binding site or very close matches to it. Moreover, these target segments include the 13 experimentally verified p53 binding regions of Cawley et al. (2004), as well as 49 additional regions that show higher hybridization signal than these 13 experimentally verified regions. PMID- 16706715 TI - Protein image alignment via piecewise affine transformations. AB - We present a new approach for aligning families of 2D gels. Instead of choosing one of the gels as reference and performing a pairwise alignment, we construct an ideal gel that is representative of the entire family and obtain a set of piecewise affine transformations that optimally align each gel of the family to the ideal gel. The coefficients defining the transformations as well as the ideal landmarks are obtained as the solution of a large-scale quadratic programming problem that can be solved efficiently by interior-point methods. PMID- 16706716 TI - Modular DAG-RNN architectures for assembling coarse protein structures. AB - We develop and test machine learning methods for the prediction of coarse 3D protein structures, where a protein is represented by a set of rigid rods associated with its secondary structure elements (alpha-helices and beta strands). First, we employ cascades of recursive neural networks derived from graphical models to predict the relative placements of segments. These are represented as discretized distance and angle maps, and the discretization levels are statistically inferred from a large and curated dataset. Coarse 3D folds of proteins are then assembled starting from topological information predicted in the first stage. Reconstruction is carried out by minimizing a cost function taking the form of a purely geometrical potential. We show that the proposed architecture outperforms simpler alternatives and can accurately predict binary and multiclass coarse maps. The reconstruction procedure proves to be fast and often leads to topologically correct coarse structures that could be exploited as a starting point for various protein modeling strategies. The fully integrated rod-shaped protein builder (predictor of contact maps + reconstruction algorithm) can be accessed at http://distill.ucd.ie/. PMID- 16706717 TI - A reversible jump Markov chain Monte Carlo algorithm for bacterial promoter motifs discovery. AB - Effective probabilistic modeling approaches have been developed to find motifs of biological function in DNA sequences. However, the problem of automated model choice remains largely open and becomes more essential as the number of sequences to be analyzed is constantly increasing. Here we propose a reversible jump Markov chain Monte Carlo algorithm for estimating both parameters and model dimension of a Bayesian hidden semi-Markov model dedicated to bacterial promoter motif discovery. Bacterial promoters are complex motifs composed of two boxes separated by a spacer of variable but constrained length and occurring close to the protein translation start site. The algorithm allows simultaneous estimations of the width of the boxes, of the support size of the spacer length distribution, and of the order of the Markovian model used for the "background" nucleotide composition. The application of this method on three sequence sets points out the good behavior of the algorithm and the biological relevance of the estimated promoter motifs. PMID- 16706718 TI - Optimal sum-of-pairs multiple sequence alignment using incremental Carrillo and Lipman bounds. AB - Alignment of sequences is an important routine in various areas of science, notably molecular biology. Multiple sequence alignment is a computationally hard optimization problem which involves the consideration of different possible alignments in order to find an optimal one, given a measure of goodness of alignments. Dynamic programming algorithms are generally well suited for the search of optimal alignments, but are constrained by unwieldy space requirements for large numbers of sequences. Carrillo and Lipman devised a method that helps to reduce the search space for an optimal alignment under a sum-of-pairs measure using bounds on the scores of its pairwise projections. In this paper, we generalize Carrillo and Lipman bounds and demonstrate a novel approach for finding optimal sum-of-pairs multiple alignments that allows incremental pruning of the optimal alignment search space. This approach can result in a drastic pruning of the final search space polytope (where we search for the optimal alignment) when compared to Carrillo and Lipman's approach and hence allows many runs that are not feasible with the original method. PMID- 16706719 TI - BEAM: a beam search algorithm for the identification of cis-regulatory elements in groups of genes. AB - The identification of potential protein binding sites (cis-regulatory elements) in the upstream regions of genes is key to understanding the mechanisms that regulate gene expression. To this end, we present a simple, efficient algorithm, BEAM (beam-search enumerative algorithm for motif finding), aimed at the discovery of cis-regulatory elements in the DNA sequences upstream of a related group of genes. This algorithm dramatically limits the search space of expanded sequences, converting the problem from one that is exponential in the length of motifs sought to one that is linear. Unlike sampling algorithms, our algorithm converges and is capable of finding statistically overrepresented motifs with a low failure rate. Further, our algorithm is not dependent on the objective function or the organism used. Limiting the space of candidate motifs enables the algorithm to focus only on those motifs that are most likely to be biologically relevant and enables the algorithm to use direct evaluations of background frequencies instead of resorting to probabilistic estimates. In addition, limiting the space of candidate motifs makes it possible to use computationally expensive objective functions that are able to correctly identify biologically relevant motifs. PMID- 16706720 TI - Local gapped subforest alignment and its application in finding RNA structural motifs. AB - RNA molecules whose secondary structures contain similar substructures often have similar functions. Therefore, an important task in the study of RNA is to develop methods for discovering substructures in RNA secondary structures that occur frequently (also referred to as motifs). In this paper, we consider the problem of computing an optimal local alignment of two given labeled ordered forests F1 and F2. This problem asks for a substructure of F1 and a substructure of F2 that exhibit a high similarity. Since an RNA molecule's secondary structure can be represented as a labeled ordered forest, the problem we study has a direct application to finding potential motifs. We generalize the previously studied concept of a closed subforest to a gapped subforest and present the first algorithm for computing the optimal local gapped subforest alignment of F1 and F2. We also show that our technique can improve the time and space complexity of the previously most efficient algorithm for optimal local closed subforest alignment. Furthermore, we prove that a special case of our local gapped subforest alignment problem is equivalent to a problem known in the literature as the local sequence-structure alignment problem (lssa) and modify our main algorithm to obtain a much faster algorithm for lssa than the one previously proposed. An implementation of our algorithm is available at www.comp.nus.edu.sg/~bioinfo/LGSFAligner/. Its running time is significantly faster than the original lssa program. PMID- 16706722 TI - Exact and approximation algorithms for DNA tag set design. AB - In this paper, we propose new solution methods for designing tag sets for use in universal DNA arrays. First, we give integer linear programming formulations for two previous formalizations of the tag set design problem. We show that these formulations can be solved to optimality for problem instances of moderate size by using general purpose optimization packages and also give more scalable algorithms based on an approximation scheme for packing linear programs. Second, we note the benefits of periodic tags and establish an interesting connection between the tag design problem and the problem of packing the maximum number of vertex-disjoint directed cycles in a given graph. We show that combining a simple greedy cycle packing algorithm with a previously proposed alphabetic tree search strategy yields an increase of over 40% in the number of tags compared to previous methods. PMID- 16706721 TI - Finding motifs in protein secondary structure for use in function prediction. AB - This paper presents a novel algorithm for the discovery of biological sequence motifs. Our motivation is the prediction of gene function. We seek to discover motifs and combinations of motifs in the secondary structure of proteins for application to the understanding and prediction of functional classes. The motifs found by our algorithm allow both flexible length structural elements and flexible length gaps and can be of arbitrary length. The algorithm is based on neither top-down nor bottom-up search, but rather is dichotomic. It is also "anytime," so that fixed termination of the search is not necessary. We have applied our algorithm to yeast sequence data to discover rules predicting function classes from secondary structure. These resultant rules are informative, consistent with known biology, and a contribution to scientific knowledge. Surprisingly, the rules also demonstrate that secondary structure prediction algorithms are effective for membrane proteins and suggest that the association between secondary structure and function is stronger in membrane proteins than globular ones. We demonstrate that our algorithm can successfully predict gene function directly from predicted secondary structure; e.g., we correctly predict the gene YGL124c to be involved in the functional class "cytoplasmic and nuclear degradation." Datasets and detailed results (generated motifs, rules, evaluation on test dataset, and predictions on unknown dataset) are available at www.aber.ac.uk/compsci/Research/bio/dss/yeast.ss.mips/, and www.genepredictions.org. PMID- 16706723 TI - A stability boundary based method for finding saddle points on potential energy surfaces. AB - The task of finding saddle points on potential energy surfaces plays a crucial role in understanding the dynamics of a micromolecule as well as in studying the folding pathways of macromolecules like proteins. The problem of finding the saddle points on a high dimensional potential energy surface is transformed into the problem of finding decomposition points of its corresponding nonlinear dynamical system. This paper introduces a new method based on TRUST-TECH (TRansformation Under STability reTained Equilibria CHaracterization) to compute saddle points on potential energy surfaces using stability boundaries. Our method explores the dynamic and geometric characteristics of stability boundaries of a nonlinear dynamical system. A novel trajectory adjustment procedure is used to trace the stability boundary. Our method was successful in finding the saddle points on different potential energy surfaces of various dimensions. A simplified version of the algorithm has also been used to find the saddle points of symmetric systems with the help of some analytical knowledge. The main advantages and effectiveness of the method are clearly illustrated with some examples. Promising results of our method are shown on various problems with varied degrees of freedom. PMID- 16706724 TI - Reconstructing ancestral haplotypes with a dictionary model. AB - We propose a dictionary model for haplotypes. According to the model, a haplotype is constructed by randomly concatenating haplotype segments from a given dictionary of segments. A haplotype block is defined as a set of haplotype segments that begin and end with the same pair of markers. In this framework, haplotype blocks can overlap, and the model provides a setting for testing the accuracy of simpler models invoking only nonoverlapping blocks. Each haplotype segment in a dictionary has an assigned probability and alternate spellings that account for genotyping errors and mutation. The model also allows for missing data, unphased genotypes, and prior distribution of parameters. Likelihood evaluations rely on forward and backward recurrences similar to the ones encountered in hidden Markov models. Parameter estimation is carried out with an EM algorithm. The search for the optimal dictionary is particularly difficult because of the variable dimension of the model space. We define a minimum description length criteria to evaluate each dictionary and use a combination of greedy search and careful initialization to select a best dictionary for a given dataset. Application of the model to simulated data gives encouraging results. In a real dataset, we are able to reconstruct a parsimonious dictionary that captures patterns of linkage disequilibrium well. PMID- 16706725 TI - The loss in power when the test of differential expression is performed under a wrong scale. AB - One of the most common and important goals of microarray studies is to identify genes that are differentially expressed between cells of different conditions. T test and ANOVA models on the expression data are common practices to gauge the significance of the observed difference in expression levels. Transformation of the microarray data is often applied in order to satisfy the model assumptions being entertained. However, the distributional properties of the expression are gene specific, and it is impractical to find a single transformation that is universally optimal for all the genes. This difficulty results in the situation that some genes have to violate the assumptions of the model (e.g., homogeneity in variance, normality). It is thus the interest of this paper to evaluate the impact on the inference of differential expression when the test is performed under an inappropriate scale. Particularly, we quantitatively assess the loss of power when the test is performed under a wrong scale. Normal distribution and log normal distribution of the expression data are considered. The loss in power is investigated in two scenarios: a transformation is misused, or a transformation fails to be applied. Log transformation and power transformation are particularly considered due to the fact that Box-Cox types of transformation are commonly used in practice. The impact of using a wrong scale is investigated analytically and based on simulations. The loss in power is assessed both as a function of the degree to which the assumptions are violated and as a function of the effect size. Simulations are conducted to quantitatively assess the power loss when tests are performed under a wrong scale. A public experimental microarray dataset is used to illustrate the impact of transformation on the results of testing differential expression. The results show that the loss of power is a function of CV and fold-change (effect size). The loss in power depends on the true model and on how severely the assumptions are violated. PMID- 16706726 TI - On methods for gene function scoring as a means of facilitating the interpretation of microarray results. AB - As gene annotation databases continue to evolve and improve, it has become feasible to incorporate the functional and pathway information about genes, available in these databases into the analysis of gene expression data, for a better understanding of the underlying mechanisms. A few methods have been proposed in the literature to formally convert individual gene results into gene function results. In this paper, we will compare the various methods, propose and examine some new ones, and offer a structured approach to incorporating gene function or pathway information into the analysis of expression data. We study the performance of the various methods and also compare them on real data, using a case study from the toxicogenomics area. Our results show that the approaches based on gene function scores yield a different, and functionally more interpretable, array of genes than methods that rely solely on individual gene scores. They also suggest that functional class scoring methods appear to perform better and more consistently than overrepresentation analysis and distributional score methods. PMID- 16706727 TI - How scale-free are biological networks. AB - The concept of scale-free network has emerged as a powerful unifying paradigm in the study of complex systems in biology and in physical and social studies. Metabolic, protein, and gene interaction networks have been reported to exhibit scale-free behavior based on the analysis of the distribution of the number of connections of the network nodes. Here we study 10 published datasets of various biological interactions and perform goodness-of-fit tests to determine whether the given data is drawn from the power-law distribution. Our analysis did not identify a single interaction network that has a nonzero probability of being drawn from the power-law distribution. PMID- 16706728 TI - Maximum likelihood molecular clock comb: analytic solutions. AB - Maximum likelihood (ML) is increasingly used as an optimality criterion for selecting evolutionary trees, but finding the global optimum is a hard computational task. Because no general analytic solution is known, numeric techniques such as hill climbing or expectation maximization (EM), are used in order to find optimal parameters for a given tree. So far, analytic solutions were derived only for the simplest model--three taxa, two state characters, under a molecular clock. Four taxa rooted trees have two topologies--the fork (two subtrees with two leaves each) and the comb (one subtree with three leaves, the other with a single leaf). In a previous work, we devised a closed form analytic solution for the ML molecular clock fork. In this work, we extend the state of the art in the area of analytic solutions ML trees to the family of all four taxa trees under the molecular clock assumption. The change from the fork topology to the comb incurs a major increase in the complexity of the underlying algebraic system and requires novel techniques and approaches. We combine the ultrametric properties of molecular clock trees with the Hadamard conjugation to derive a number of topology dependent identities. Employing these identities, we substantially simplify the system of polynomial equations. We finally use tools from algebraic geometry (e.g., Grobner bases, ideal saturation, resultants) and employ symbolic algebra software to obtain analytic solutions for the comb. We show that in contrast to the fork, the comb has no closed form solutions (expressed by radicals in the input data). In general, four taxa trees can have multiple ML points. In contrast, we can now prove that under the molecular clock assumption, the comb has a unique (local and global) ML point. (Such uniqueness was previously shown for the fork.). PMID- 16706729 TI - Bayesian sequential inference for stochastic kinetic biochemical network models. AB - As postgenomic biology becomes more predictive, the ability to infer rate parameters of genetic and biochemical networks will become increasingly important. In this paper, we explore the Bayesian estimation of stochastic kinetic rate constants governing dynamic models of intracellular processes. The underlying model is replaced by a diffusion approximation where a noise term represents intrinsic stochastic behavior and the model is identified using discrete-time (and often incomplete) data that is subject to measurement error. Sequential MCMC methods are then used to sample the model parameters on-line in several data-poor contexts. The methodology is illustrated by applying it to the estimation of parameters in a simple prokaryotic auto-regulatory gene network. PMID- 16706730 TI - Towards a simple, saliva-based test for the detection of oral cancer 'oral fluid (saliva), which is the mirror of the body, is a perfect medium to be explored for health and disease surveillance'. PMID- 16706732 TI - AmpliChip CYP450 test: personalized medicine has arrived in psychiatry. AB - The US FDA has granted market approval for the first pharmacogenetic test using a DNA microarray, the AmpliChip CYP450, which genotypes cytochrome P450 (CYP)2D6 and CYP2C19. The test uses software to predict phenotypes and tests for 27 CYP2D6 alleles, including the deletions and duplications, and three CYP2C19 alleles. Other DNA microarray platforms are being developed for CYP testing, but none have been completely developed or approved by the FDA to date. The differences between an implementation of pharmacogenetic tests centered on the individual and implementation using a public health approach are discussed. In this review, the major obstacles to the wide implementation of pharmacogenetic testing in the clinical environment are summarized. PMID- 16706733 TI - Microelectronic array system for molecular diagnostic genotyping: Nanogen NanoChip 400 and molecular biology workstation. AB - Hundreds of gene mutations responsible for Mendelian disorders are currently tested in the clinical laboratory for pre- and postnatal diagnosis, carrier screening and presymptomatic testing. Since human genetic research is currently focused on determining the etiology of complex diseases, including heart disease, diabetes and neuropsychiatric traits, laboratorians will genotype increasing numbers of clinically relevant loci in the future. This will require accurate, high-throughput and cost-effective genotyping platforms, such as the DNA microarray. The Nanogen NanoChip platforms employ hybridization-based technology, using fluorescent detection and electronic control of the target or probe, to obtain clear genotype signal relative to background, and increased flexibility relative to similar chip-based single nucleotide polymorphism genotyping platforms. The scope of this review is intended to describe the operating principle, chips and instrumentation, analyte-specific reagents, published assay protocols, assay development, and clinical use of the NanoChip platforms. It is beyond the scope of this review to describe the use of NanoChip platforms in basic research, and to compare it against all available clinical single nucleotide polymorphism genotyping applications and platforms. PMID- 16706734 TI - DualChip microarray as a new tool in cancer research. AB - Over the last 5 years, the emergence of gene expression profiling using high density DNA microarrays led to a better understanding of tumor development and identified new prognostic markers. However, high-density microarrays failed to leap from the researcher's bench to the clinical practice due to their cost, data management and lack of standardization. DualChip low-density DNA microarrays were developed as a new flexible tool that is able to reliably quantify the expression of a limited number of genes of clinical relevance. This review will illustrate how DualChip technology can be applied to tumor diagnosis and tumor-acquired drug resistance. PMID- 16706735 TI - Nanotechnology for cancer diagnostics: promises and challenges. AB - Despite recent progress in the treatment of cancer, the majority of cases are still diagnosed only after tumors have metastasized, leaving the patient with a grim prognosis. However, there may be an opportunity to drastically reduce the burden of cancer, if the disease can be detected early enough. Nanotechnology is in a unique position to transform cancer diagnostics and to produce a new generation of biosensors and medical imaging techniques with higher sensitivity and precision of recognition. This review examines the in vitro and in vivo diagnostic applications of nanoparticles, and other nanodevices that are likely to have an impact on the field in the future. Future developments that may lead to the realization of multifunctional detection and treatment nanoparticle platforms are also discussed. PMID- 16706736 TI - Challenges of SNP genotyping and genetic variation: its future role in diagnosis and treatment of cancer. AB - Thorough annotation of common germline genetic variation in the human genome has generated a foundation for the investigation of the contribution of genetics to the etiology and pathogenesis of cancer. For many malignancies, it has become increasingly apparent that numerous alleles, with small-to-moderate effects, additively contribute to cancer susceptibility. The most common genetic variant in the genome, the single nucleotide polymorphism, is of special interest for the study of susceptibility to and protection from cancer. Similarly, intense effort has focused on genetic variants that can predict either response or toxicity to therapeutic interventions. This review discusses the challenges and prospects of genetic association studies in cancer research. On the basis of recent changes in genomics and high-throughput genotyping platforms, future genetic findings of association studies could impact clinical care and public health screening. PMID- 16706737 TI - Impact of disseminated tumor cells in gastrointestinal cancer. AB - The detection of epithelial cells by sensitive immunological and molecular methods in blood, lymph nodes or bone marrow of gastrointestinal cancer patients may open a new approach to clinical metastasis research. The phenotypic and genomic characterization of these cells is of great value in the prediction of the further course of the disease and the monitoring of response to treatment. In addition, the role of ultrastaging in blood, lymph nodes and bone marrow of cancer patients for the indication of multimodal therapy is discussed in this review. The impact of prognostic or predictive factors for new treatment protocols in patients with gastrointestinal cancer was evaluated as well as the correlation with clinical factors. PMID- 16706738 TI - Role of genes, the environment and their interactions in the etiology of inflammatory bowel diseases. AB - Few of the studied genes demonstrate association with inflammatory bowel disease (IBD). Three mutations in the nucleotide-binding oligomerization domain 2 gene have consistently shown to be independent risk factors for Crohn's disease, but none of the alleles exhibited high sensitivity or specificity for IBD. Linkage analysis implicated several loci on various chromosomes, and epistasis has been demonstrated. The etiopathogenesis of IBD remains unknown, and environmental contribution to their pathogenesis is evident from genetic studies that demonstrated incomplete monozygotic twins concordandance rate for both Crohn's and ulcerative colitis. Smoking has shown an opposite effect on disease phenotype, with an adverse effect on disease course for Crohn's disease, but a slight beneficial effect in ulcerative colitis. The contribution of infectious agents to susceptibility to IBD appears to be strong. However, the role of nutrition on the etiology and therapy of IBD is not clear. Inconsistencies in environmental risk factors could be due to gene-environment interactions, making it essential to study the role of genetics and environmental contribution to the etiopathology of IBD. Transgenic or knockout mice, such as interleukin-10(-/-), T cell receptor alpha(-/-), Galphai(2) (-/-) and N-cadherin(-/-), develop colitis like inflammation similar to humans. Therefore, animal models must be further studied to explore mechanistic interactions. PMID- 16706739 TI - Congenital long QT syndromes: clinical features, molecular genetics and genetic testing. AB - Congenital long QT syndrome (LQTS) is a primary electrical disease characterized by a prolonged QT interval in the surface electrocardiogram and increased predisposition to a typical polymorphic ventricular tachycardia, termed Torsade de Pointes. Most patients with LQTS are asymptomatic and are diagnosed incidentally based on an electrocardiogram. Symptomatic patients may suffer from severe cardiac events, such as syncope and/or sudden cardiac death. Autosomal dominant forms are caused by heterozygous mutations in genes encoding the components of the ion channels. The autosomal recessive form with congenital deafness is also known as Jervell and Lang-Nielsen syndrome. It is caused by homozygous mutations or certain compound heterozygous mutations. Depending on the genetic defects, there are differences in the age of onset, severity of symptoms, and number of cardiac events and event triggers. With advances in gene technology, it is now feasible to perform genetic testing for LQTS, especially for those with family history. Identification of the mutation will lead to better management of symptoms and more targeted treatment, depending on the underlying genetic defect, resulting in a reduction of mortality and cardiac events. PMID- 16706740 TI - Hereditary sensorineural hearing loss: advances in molecular genetics and mutation analysis. AB - Hearing loss has a genetic etiology in the majority of cases and is very common. The universal newborn hearing screening program, together with remarkable recent progress in the characterization of genes associated with the function of hearing, have resulted in increased demand and exciting possibilities of detecting the molecular basis of hereditary hearing loss through DNA testing. Future molecular diagnostic assays are expected to offer a greater variety of gene-specific tests, as well as combined mutation panels, which will aid in the management of the impressive genetic heterogeneity observed in hereditary hearing loss, especially in individuals with nonsyndromic forms. This review addresses the genetics of hearing loss, discusses the most commonly offered genetic assays for nonsyndromic hearing loss, with advantages and limitations, proposes a practical testing algorithm, and highlights current developments. PMID- 16706741 TI - Biomarkers for amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is the most common form of motor neuron disease. ALS is a fatal neurodegenerative disease and clinical diagnosis typically takes many months to complete. Early disease diagnosis through the use of biomarkers may aid in correct clinical management of patients and possibly delay time to ventilator and morbidity. This review explores the progress of biomarker discovery efforts for ALS and the many challenges that remain. Included are different technologies utilized in biomarker discovery efforts (proteomic, genomic and metabolomic) and putative biomarkers uncovered using these techniques. These studies have discovered genetic mutations leading to familial forms of ALS, and specific protein alterations that occur in biological fluids (cerebrospinal fluid and blood) and/or tissues of ALS subjects. More recent high throughput technologies have revealed panels of proteomic or metabolic biomarkers that can discriminate between ALS and control groups. The identification of disease-specific biomarkers will provide opportunities to develop early diagnostic measures as well as surrogate markers to monitor disease progression and test drug efficacy in clinical trials. PMID- 16706742 TI - Screening of HIV infection: role of molecular and immunological assays. AB - Due to technical improvements and new developments of immunological assays, the reliability of serological laboratory diagnosis of HIV infection has improved considerably and the residual risk, due to the diagnostic window for transfusion transmitted HIV, has been reduced significantly. Through the addition of nucleic acid amplification tests (NAT) to blood donor screening, the residual risk can de further decreased by up to 50%, depending on the sensitivity of the NAT protocol and whether individual or pooled blood donations are screened. In-house and commercially available NAT have been implemented in blood banks as HIV only or multiplexed HIV and hepatitis B or C virus assays. As an alternative to separate antigen and antibody screening, combined fourth-generation assays have been developed in 1997, and have achieved a high degree of sensitivity and specificity. Thus, they can replace stand-alone antigen and third-generation antibody assays. While they are used in the routine diagnostics of HIV infection in many countries throughout the world, they probably represent no alternative for NAT in blood-donor screening in industrialized countries. In the next few years, technical improvements will further simplify NAT screening. While there is still some potential to improve the detection threshold of NAT, the sensitivity of the antigen module of fourth-generation assays (a lowest concentration of 3-5 pg of p24 antigen) is probably very close to its technical limit. PMID- 16706743 TI - New tools and emerging technologies for the diagnosis of tuberculosis: part I. Latent tuberculosis. AB - Nearly a third of the world's population is estimated to be infected with Mycobacterium tuberculosis. This enormous pool of latently infected individuals poses a major hurdle for global tuberculosis (TB) control. Currently, diagnosis of latent TB infection (LTBI) relies on the tuberculin skin test (TST), a century old test with known limitations. In this review, the first of a two-part series on new tools for TB diagnosis, recent advances in the diagnosis of LTBI are described. The biggest advance in recent years has been the development of in vitro T-cell-based interferon-gamma release assays (IGRAs) that use antigens more specific to M. tuberculosis than the purified protein derivative used in the TST. Available research evidence on IGRAs suggests they have higher specificity than TST, better correlation with surrogate markers of exposure to M. tuberculosis in low-incidence settings, and less cross-reactivity due to BCG vaccination than the TST. IGRAs also appear to be at least as sensitive as the purified protein derivative-based TST for active TB. In the absence of a gold standard for LTBI, sensitivity and specificity for LTBI are not well defined. Besides high specificity, other potential advantages of IGRAs include logistical convenience, avoidance of poorly reproducible measurements, such as skin induration, need for fewer patient visits and the ability to perform serial testing without inducing the boosting phenomenon. Overall, due to its high specificity, IGRAs may be useful in low-endemic, high-income settings where cross-reactivity due to BCG might adversely impact the utility of TST. However, despite the growing evidence supporting the use of IGRAs, several unresolved and unexplained issues remain. The review concludes by highlighting areas where evidence is lacking, and provides an agenda for future research. Active TB and drug resistance are discussed in Part II; 423-432 of this issue. PMID- 16706744 TI - New tools and emerging technologies for the diagnosis of tuberculosis: part II. Active tuberculosis and drug resistance. AB - Tuberculosis (TB) is one of the world's most important infectious causes of morbidity and mortality among adults. Between 8 and 9 million develop TB disease, and approximately 2 million die from TB each year. Despite this enormous global burden, case detection rates are low, posing major hurdles for TB control. Conventional TB diagnosis continues to rely on smear microscopy, culture and chest radiography. These tests have known limitations. Conventional tests for detection of drug resistance are slow, tedious and difficult to perform in field conditions. This second half of a two-part review series on new tools for TB diagnosis describes recent advances and emerging technologies in the diagnosis of active disease, and detection of drug resistance. For diagnosis, new tools include newer versions of nucleic acid amplification tests, immune-based assays, skin patch test and rapid culture systems. For drug resistance, new tools include line-probe assays, bacteriophage-based assays, molecular beacons and microscopic observation drug susceptibility assay. Although the ideal test for TB is still not in sight, substantial progress has been made in the past decade. With the resurgence of interest in the development of new tools for TB control, it is likely that the next decade will see greater progress and tangible benefits. However, the challenge will be to ensure that new tools undergo rigorous evaluations in field conditions, and also to make sure that benefits of promising new tools actually reach the populations in developing countries that need them most. Latent TB is discussed in Part I; 413-422 of this issue. PMID- 16706745 TI - Next revolution in the molecular theranostics of infectious diseases: microfabricated systems for personalized medicine. AB - The molecular diagnosis of infectious diseases is currently going through a revolution sustained by the regulatory approval of amplification tests that have been shown to be equivalent or superior to existing gold standard methods. The recent approval of a microarray system for the pharmacogenomic profiling of cytochrome P450-mediated drug metabolism is paving the way to novel, rapid, sensitive, robust and economical microfabricated systems for point-of-care diagnostics, which are utilized closer and closer to the patient's bedside. These systems will enable the multiparametric genetic evaluation of several medical conditions, including infectious diseases. This forecoming revolution will position molecular theranostics in a broader integrated view of personalized medicine, which exploits genetic information from microbes and human hosts to optimize patient management and disease treatment. PMID- 16706746 TI - Application of oligonucleotide arrays to high-content genetic analysis. AB - The scope of single nucleotide polymorphism genotyping for genetic association studies has expanded recently from the use of relatively small numbers of candidate genes and markers, to include hypothesis-free, whole-genome approaches using hundreds of thousands of polymorphisms. The ability to perform such large scale association studies has been dependant on the development of highly parallel and cost-effective genotyping platforms, of which those based on oligonucleotide arrays have proved to be the most scalable and widely adopted. It is to be expected that the new array-based genotyping methods will not only greatly expand the scope of genetic studies, but, as further content is added to arrays, will also form part of an integrated set of DNA, RNA and proteomic analyses enabling the detailed, multilayered study of complex disease-linked phenotypes. PMID- 16706747 TI - Transcriptome amplification methods in gene expression profiling. AB - The increasing use of microarray expression profiling to study the molecular biology of cancer and the cellular physiology of difficult-to-isolate cell types has led to a need for methods that accurately and precisely amplify small quantities of RNA. The purpose of this review is to provide an overview of the existing methods for transcriptome amplification and to define the parameters for comparing different amplification methods. The authors propose a standardized protocol for the assessment and evaluation of amplification methods, focusing on a new whole-transcriptome amplification kit, which amplifies total RNA into cDNA fragments. Reproducibility and reliability of the method were analyzed and discussed using both quantitative real-time PCR and a high-density oligonucleotide microarray platform. PMID- 16706748 TI - Improving reliability and performance of DNA microarrays. AB - A great many platforms and versions of the microarray technology, with different characteristics and applications, have been developed. This review will describe some key issues in reliability and performance with the two most commonly used platforms for gene expression analysis, in situ-synthesized oligonucleotide microarrays or GeneChips and spotted microarrays. Some recent advances and new applications within the field will be mentioned briefly. PMID- 16706749 TI - Bubble dynamics involved in ultrasonic imaging. AB - In clinical ultrasound, blood cells cannot be differentiated from surrounding tissue, due to the low acoustic impedance difference between blood cells and their surroundings. Resonant gas bubbles introduced in the bloodstream are ideal markers, if rapid dissolution can be prevented. Ultrasound contrast agents consist of microscopically small bubbles encapsulated by an elastic shell. These microbubbles oscillate upon ultrasound insonification. Microbubbles with thin lipid shells have demonstrated highly nonlinear behavior. To enhance diagnostic ultrasound imaging techniques and to explore therapeutic applications, these medical microbubbles have been modeled. Several detection techniques have been proposed to improve the detectability of the microbubbles. A new generation of contrast agents, with special targeting ligands attached to the shells, may assist the imaging of nonphysical properties of target tissue. Owing to microbubble-based contrast agents, ultrasound is becoming an even more important technique in clinical diagnostics. PMID- 16706750 TI - Lipid raft components cholesterol and sphingomyelin increase H+/OH- permeability of phosphatidylcholine membranes. AB - H+/OH- permeation through lipid bilayers occurs at anomalously high rates and the determinants of proton flux through membranes are poorly understood. Since all life depends on proton gradients, it is important to develop a greater understanding of proton leak phenomena. We have used stopped-flow fluorimetry to probe the influence of two lipid raft components, chol (cholesterol) and SM (sphingomyelin), on H+/OH- and water permeability. Increasing the concentrations of both lipids in POPC (palmitoyl-2-oleoyl phosphatidylcholine) liposomes decreased water permeability in a concentration-dependent manner, an effect that correlated with increased lipid order. Surprisingly, proton flux was increased by increasing the concentration of chol and SM. The chol effect was complex with molar concentrations of 17.9, 33 and 45.7% giving 2.8-fold (P<0.01), 2.2-fold (P<0.001) and 5.1-fold (P<0.001) increases in H+/OH- permeability from a baseline of 2.4x10(-2) cm/s. SM at 10 mole% effected a 2.8-fold increase (P<0.01), whereas 20 and 30 mole% enhanced permeability by 3.6-fold (P<0.05) and 4.1-fold respectively (P<0.05). Supplementing membranes containing chol with SM did not enhance H+/OH- permeability. Of interest was the finding that chol addition to soya-bean lipids decreased H+/OH- permeability, consistent with an earlier report [Ira and Krishnamoorthy (2001) J. Phys. Chem. B 105, 1484-1488]. We speculate that the presence of proton carriers in crude lipid extracts might contribute to this result. We conclude that (i) chol and SM specifically and independently increase rates of proton permeation in POPC bilayers, (ii) domains enriched in these lipids or domain interfaces may represent regions with high H+/OH- conductivity, (iii) H+/OH- fluxes are not governed by lipid order and (iv) chol can inhibit or promote H+/OH- permeability depending on the total lipid environment. Theories of proton permeation are discussed in the light of these results. PMID- 16706752 TI - New comprehension of the apicoplast of Sarcocystis by transmission electron tomography. AB - BACKGROUND INFORMATION: Apicomplexan parasites (like Plasmodium, Toxoplasma, Eimeria and Sarcocystis) contain a distinctive organelle, the apicoplast, acquired by a secondary endosymbiotic process analogous to chloroplasts and mitochondria. The apicoplast is essential for long-term survival of the parasite. This prokaryotic origin implies that molecular and metabolic processes in the apicoplast differ from those of the eukaryotic host cells and therefore offer options for specific chemotherapeutic treatment. We studied the apicoplast in high-pressure frozen and freeze-substituted cysts of Sarcocystis sp. from roe deer (Capreolus capreolus) to get better insight in apicoplast morphology. RESULTS AND CONCLUSIONS: We observed that the apicoplast contains four continuous membranes. The two inner membranes have a circular shape with a constant distance from each other and large-sized protein complexes are located between them. The two outer membranes have irregular shapes. The periplastid membrane also contains large-sized protein complexes, while the outer membrane displays protuberances into the parasite cytoplasm. In addition, it is closely associated with the endoplasmic reticulum by 'contact sites'. PMID- 16706751 TI - Polyamine-modulated c-Myc expression in normal intestinal epithelial cells regulates p21Cip1 transcription through a proximal promoter region. AB - Maintenance of intestinal mucosal epithelial integrity requires cellular polyamines that regulate expression of various genes involved in cell proliferation, growth arrest and apoptosis. Our previous studies have shown that polyamines are essential for expression of the c-myc gene and that polyamine induced c-Myc plays a critical role in stimulation of normal IEC (intestinal epithelial cell) proliferation, but the exact downstream targets of induced c-Myc are still unclear. The p21Cip1 protein is a major player in cell cycle control, which is primarily regulated at the transcriptional level. The current study was designed to determine whether induced c-Myc stimulates normal IEC proliferation by repressing p21Cip1 transcription following up-regulation of polyamines. Overexpression of the ODC (ornithine decarboxylase) gene increased levels of cellular polyamines, induced c-Myc expression and inhibited p21Cip1 transcription, as indicated by repression of p21Cip1 promoter activity and a decrease in p21Cip1 protein levels. In contrast, depletion of cellular polyamines by inhibiting ODC enzyme activity with alpha-difluoromethylornithine decreased c Myc, but increased p21Cip1 transcription. Ectopic expression of wild-type c-myc not only inhibited basal levels of p21Cip1 transcription in control cells, but also prevented increased p21Cip1 in polyamine-deficient cells. Experiments using different p21Cip1 promoter mutants showed that transcriptional repression of p21Cip1 by c-Myc was mediated through Miz-1- and Sp1-binding sites within the proximal region of the p21Cip1 promoter in normal IECs. These findings confirm that p21Cip1 is one of the direct mediators of induced c-Myc following increased polyamines and that p21Cip1 repression by c-Myc is implicated in stimulation of normal IEC proliferation. PMID- 16706753 TI - Near [corrected] painless, nonablative, immediate skin contraction induced by low fluence irradiation with new infrared device: a report of 25 patients. AB - BACKGROUND: Nonablative radiofrequency (NARF) has been the only method for producing noninvasive skin tightening. Nevertheless, significant pain during the procedure is an important downside of this technology. A new nonablative medical device, Titan (Cutera, Inc., Brisbane, CA, USA), capable of fluences much lower than those possible with NARF, was tested as a less painful alternative. OBJECTIVES: To produce skin contraction leading to lifting of eyebrows and/or improvement of lower face and neck skin laxity using fluences below pain levels. PATIENTS AND METHODS: Twenty-five patients were treated. Standardized photographs were obtained preoperatively, after a few days, a few weeks, and up to 12 months after the procedure. RESULTS: Immediate changes were obtained in 22 of 25 patients. Examination of photographs revealed that the initial improvement was maintained throughout the follow-up period. CONCLUSION: Immediate true skin contraction persisting through the immediate, intermediate, and long-term follow up was found in the vast majority of patients in this group. Edema as an artifact simulating immediate improvement was excluded by serial photographs taken during the follow-up period. Skin contraction occurred at low fluences, below the threshold of pain. This, to the best of our knowledge, has not been previously described in the medical literature. PMID- 16706754 TI - Phenol-castor oil: modified peel for dermal melasma. AB - BACKGROUND: Deep type of melasma is difficult to treat. Various compositions for peeling have been formulated to accomplish this. OBJECTIVE: Our purpose was to determine the safe use of the new formula, which was composed of phenol and castor oil. METHODS: Different concentrations of phenol and castor oil were mixed in five formulations, and the solutions were tested on the forearms of 10 volunteers. The application time was 1 minute, different from that of the original phenol-croton oil. Formula 4 was found to cause a high effect of peeling with least side effects and was used to treat deep facial melasma in 30 patients. RESULTS: Higher concentrations of castor oil produced more inflammation when mixed with phenol. Formula 4 was used to clear two cases of melasma in 1 week despite no effect from prior use of a modified antimelasma cream, while some degree of fading was noted in other cases. The total number of different melasma indices was reduced from 206.4 to 91.2 in 30 patients. Hyperpigmentation was noted in five cases and hypopigmentation in one case. Neither scar formation nor cardiac arrhythmia was observed. CONCLUSION: A phenol-castor oil peel with an application time of 1 minute reduces pigments in the deep type of melasma with least side effects and a high efficiency when combined with an antimelasma cream. The safe formula and the technique of application are introduced. PMID- 16706755 TI - Improvement in the appearance of wrinkles with topical transforming growth factor beta(1) and l-ascorbic acid. AB - BACKGROUND AND OBJECTIVE: Facial rhytides are a common cosmetic concern. Surgical treatment effects dramatic improvement; however, the associated risk and cost may be prohibitive. Recent focus is on developing topical products containing biologically active ingredients for at-home therapy. Our study examines the effects of a topical cream containing transforming growth factor-beta(1) (TGF beta(1)), l-ascorbic acid, and Cimicifuga racemosa extract (CRS) (Topix Pharmaceuticals, North Amityville, NY, USA). MATERIALS AND METHODS: In the first arm of the study, 12 subjects were randomized to apply CRS to the left or right side of their face and a cream containing l-ascorbic acid and C. racemosa in silicone base (vitamin C [Vit C]) to the contralateral side twice daily for 3 months. In the second arm of the study, 20 subjects were randomized to apply CRS to the left or right side of their face and Tissue Nutrient Solution Recovery Complex (TNS) (SkinMedica, Carlsbad, CA, USA), a product containing a variety of growth factors including VEGF, PDGF-A, G-CSF, HGF, IL-6, IL-8, and TGF-beta(1) (Nouricel-MD) without l-ascorbic acid, C. racemosa extract, or silicone base, to the contralateral side of their face twice daily for 3 months. Digital photographs were scored by study-blinded physicians, and self-assessments were completed by all subjects at baseline and at the conclusion of the protocol. RESULTS: CRS and TNS were well tolerated, and all subjects completed the 3-month protocol for the CRS versus Vit C arm of the study. Physicians rated success in facial wrinkling scores on the CRS-treated side of the face for 27 of 31 subjects. Responders showed, on average, 21.7% improvement in physician-rated wrinkle scores. The mean improvement in the group of 31 patients as a whole was 12%. Eighteen of 31 subjects reported a noticeable improvement on their CRS treated side. Both CRS and TNS demonstrate significant success between baseline and 3-month scores, and both growth factor products are superior to Vit C. Patients preferred CRS over TNS. CONCLUSION: CRS is effective in minimizing the appearance of facial rhytides. The success of the CRS product is largely attributable to the incorporation of TGF-beta(1). PMID- 16706756 TI - Localized intradermal microinjection of tranexamic acid for treatment of melasma in Asian patients: a preliminary clinical trial. AB - BACKGROUND: Melasma is a common cosmetic problem among Asians. While various treatments are currently being used, there is no entirely satisfactory treatment. It was recently reported that the topical plasmin inhibitor is an effective treatment for ultraviolet-induced hyperpigmentation. OBJECTIVE: Because there are no studies assessing the efficacy and safety of localized microinjection of tranexamic acid (TA) for the treatment of melasma, we conducted a pilot study to evaluate the efficacy and side effects of this potentially new method for the treatment of melasma in Korean women. METHODS: A total of 100 women with melasma, after written consent, were enrolled for a prospective open pilot study of 12 weeks. After applying topical anesthesia, 0.05 mL TA (4 mg/mL) was injected intradermally into the melasma lesion at 1 cm intervals by using a 0.5 mL insulin syringe with a 30-gauge needle. This was repeated weekly for 12 weeks. A clinical investigator evaluated the results by using the Melasma Area and Severity Index (MASI) at baseline and at 4, 8, and 12 weeks. The patient satisfaction questionnaire was documented at 12 weeks. Safety evaluations were performed at each follow-up visit. RESULTS: Eighty-five patients completed the trial. A significant decrease in the MASI from baseline to 8 and 12 weeks was observed (13.22+/-3.02 vs 9.02+/-2.62 at week 8 and vs. 7.57+/-2.54 at week 12; p<.05 for both). The patients' self-assessment of melasma improvement was as follows: 8 of 85 patients (9.4%) rated as good (51-75% lightening), 65 patients (76.5%) as fair (26-50% lightening), and 12 patients (14.1%) as poor (0-25% lightening). Side effects were minimal and all the patients tolerated the treatment well. CONCLUSION: Based on these results, we suggest that the intralesional localized microinjection of TA acid can be used as a potentially new, effective, and safe therapeutic modality for the treatment of melasma. PMID- 16706757 TI - Extended monitoring of hemostatic activation after varicose vein surgery under general anesthesia. AB - BACKGROUND: Postoperative heparin prophylaxis after stripping of the long saphenous vein is a matter of controversial discussion, and practices vary by surgeon and country. OBJECTIVE: The aim of this study was to assess the extent of hypercoagulability by continued monitoring of activation markers of coagulation and fibrinolysis for a period of 3 weeks after stripping of the long saphenous vein and concomitant phlebectomy. METHODS: Including 21 patients, the following markers were measured preoperatively and on postoperative day 1, 2, 3, 7, 14, and 21: Activation products of coagulation: thrombin-antithrombin complex (TAT), thrombus precursor protein (TPP), and prothrombin-fragment F1+2 (F1+2), and markers of fibrinolysis: plasmin-alpha(2)-antiplasmin complexes (PAP), D-Dimer, tissue plasminogen activator (t-PA) antigen, and plasminogen activator inhibitor 1 (PAI-1) antigen. RESULTS: TAT levels increased significantly until day 3 (p=.008) and normalized within 14 days. TPP levels increased significantly until day 7 (p=.02), decreasing to initial values within 21 days. PAP complexes increased significantly until day 2 (p=.02) reducing to baseline within the observation period. D-Dimer levels increased immediately after surgery (p<.001) until day 14 (p<.001) and returned to baseline until day 21. CONCLUSIONS: Significant hemostatic activation after varicose vein surgery was observed and persisted until 3 weeks postoperatively, indicating that heparin prophylaxis for 2 to 3 weeks is advisable for at-risk patients. PMID- 16706758 TI - Fat transplantation using fresh versus frozen fat: a side-by-side two-hand comparison pilot study. AB - BACKGROUND: Autologous fat in both fresh and frozen forms has been used for many years as a filler for various dermatologic conditions. However, it is not clear whether fat that has been frozen survives as well as, and gives aesthetic results similar to, fresh fat. The efficacy of frozen fat has been debated in the literature. OBJECTIVE: To evaluate the clinical aesthetic appearance and longevity of fresh fat versus frozen fat in a side-by-side two-hand comparison in the same patient. METHODS: Ten patients underwent fat augmentation on their hands, utilizing 10 cc of fresh fat in one hand and 10 cc of frozen fat in the contralateral hand within 17 days of fresh fat placement. Follow-up evaluation was conducted at 1, 3, and 5 months in a randomized, double-blind comparison study. Physician-determined aesthetic preference, prominence of veins, and depth of metacarpal spaces were evaluated. Photographs were taken of both hands during each patient follow-up visit. RESULTS: All three areas of physician-assessed gradation: aesthetics, vein prominence, and depth of metacarpal space were superior for the hand injected with frozen fat at 1-, 3-, and 5-month follow-up visits. CONCLUSIONS: This pilot study supports the use of autologous frozen fat for equivalent to improved results regarding longevity and aesthetic appearance versus fresh fat at 1, 3, and 5 months for fat augmentation of aging hands. PMID- 16706759 TI - Treatment of depression with botulinum toxin A: a case series. AB - BACKGROUND: Major depression is a common and serious disease that may be resistant to routine pharmacologic and psychotherapeutic treatment approaches. OBJECTIVE: To evaluate the efficacy of botulinum toxin A treatment of glabellar frown lines in treating patients with major depression, using a small open pilot trial. METHODS: Patients who met DSM-IV criteria for ongoing major depression in spite of pharmacologic or psychotherapeutic treatment were evaluated with the Beck Depression Inventory II (BDI-II) before receiving botulinum toxin A to their glabellar frown lines. Two months later, all patients were re-evaluated clinically and with the BDI-II. RESULTS: Ten depressed patients were treated with botulinum toxin A, and 9 of 10 patients were no longer depressed 2 months after treatment. The tenth patient had an improvement in mood. CONCLUSION: To our knowledge, these are the first reported cases of depression treated with botulinum toxin A. PMID- 16706760 TI - Orientation of multi-hair follicles in nonbald men: perpendicular versus parallel. AB - BACKGROUND: There has been much debate about the orientation of multi-hair grafts in order to obtain an appearance of fullness after the hair transplantation. OBJECTIVES: To analyze natural orientation of multi-hair follicles and the influence of hair flow from the crown swirl. METHODS: Natural orientation of follicles was analyzed microscopically in 100 nonbald men: a clockwise whorl flow group (50 men) and a counterclockwise group (50 men). RESULTS: The majority of multi-hair follicles were oriented perpendicular to the radial line from the whorl in both the groups. The percentage of perpendicular orientation was large in the peripheral and middle scalp areas (49-79%), and it decreased in the vertex (38-49%, p<.05 or .01). The percentages of parallel, right oblique, and left oblique orientation were small in the peripheral area with a slight increase in the vertex (3-12% vs. 14-24%, p<.05 or .01). CONCLUSION: Original orientation of multi-hair follicles was mainly perpendicular to the radial line from the whorl irrelevant to the crown swirl hair flow. Multi-hair grafts transplanted perpendicularly would look similar to the natural hair follicles in the front half of the scalp. PMID- 16706761 TI - Treatment of superficial surgical wounds after removal of seborrheic keratoses: a single-blinded randomized-controlled clinical study. AB - BACKGROUND: For the treatment of superficial surgical wounds, there are a number of options, including topical antibiotic ointments, dressings, and specialized wound care materials, such as hydrocolloid dressings. OBJECTIVE: To evaluate the wound-healing activity of a commercially available hydrocolloid wound dressing (Avery H2460, Avery Dennison, Turnhout, Belgium) in comparison with a control treatment (Fucidine cream with Cutiplast sterile dressing) in superficial wounds after surgical removal of seborrheic keratoses. METHODS: In a single-blinded, randomized, controlled trial, the hydrocolloid wound dressing (Avery H2460) was compared with healing by secondary intention as a control. Sixteen patients between 18 and 80 years of age with seborrheic keratoses were enrolled. Wound healing was evaluated after 7 and 10 days and then daily until complete closure of the wound area. In 7 of 16 patients, biopsies were taken after 14 days of reepithelization. RESULTS: The hydrocolloid wound dressing (Avery H2460) induced a significantly (p<.05) faster healing (median: 8.5 days) in comparison with the control treatment (median: 10 days). The histologic investigations showed no significant differences for the investigated parameters in both groups. CONCLUSION: The faster healing in comparison with the control treatment supports the use of the hydrocolloid wound dressing (Avery H2460) for the treatment of superficial surgical wounds. PMID- 16706762 TI - Optimization of thermocautery in excisional dermatologic surgery. AB - PURPOSE: Electrosurgery is routinely used in cutaneous surgery for hemostasis. Thermocautery can be used in patients with implantable cardiac devices. This technique relies on heat without electrical current passing through the patient to produce hemostasis. The temperatures and utility of a commercially available, adjustable thermocautery unit are examined. METHODOLOGY: Tip temperature of the commercially available thermocautery unit was measured in air and tissue via a type E thermocouple (0.002 in. diameter) around the unit's tip. Time intervals of 20 to 30 seconds were recorded at device settings 1 to 9 in air and 3 to 8 on surgical patients (Institutional Review Board approval obtained). RESULTS: In vitro analysis demonstrated predictable temperatures at increasing settings in air: 350 to 900 degrees C. Analysis in vivo during surgery demonstrated similar findings. Tissue contact decreased tip temperature by approximately 50% from in vitro values, and use in a bloody field caused a further decrease in the tip temperature. CONCLUSION: The thermocautery unit examined is an effective and safe unit to achieve hemostasis. In addition, the temperature may be adjusted as opposed to hand-held units that operate at in vitro temperatures exceeding 1,400 degrees F. Hemostasis at approximately 100 to 400 degrees C provided optimal hemostasis. PMID- 16706763 TI - Prospective, randomized, blinded study of a new wound closure film versus cutaneous suture for surgical wound closure. AB - BACKGROUND: Wound closure devices include sutures, tissue adhesives, adhesive strips, and staples. Recent studies suggest no differences between various tissue adhesives and sutures for dehiscence, infection, and satisfaction when assessed by patients or surgeons. To date, no studies have investigated ClozeX, a novel adhesive strip, for closure of surgical incisions. OBJECTIVE: To compare surgical wounds repaired with ClozeX versus suture. METHODS: A prospective, randomized study was conducted, in which 15 patients with surgical incisions were allocated to closure with ClozeX on half of the wound and monofilament suture on the other half. Physician satisfaction with blinded assessment, patient satisfaction, complication rates, and closure times were recorded. RESULTS: Application with ClozeX was faster than with suture (p=.007). There were no complications in either group. Sixty-nine percent of the patients gave ClozeX a higher satisfaction score (p=.02). More physicians were satisfied with the ClozeX half than with the suture half (p=.007). CONCLUSIONS: This pilot study demonstrates ClozeX to be a safe and effective closure device. The cosmetic outcome seems to be at least as good as simple running suture. Physicians and patients were generally more satisfied with ClozeX. No difference was found in the rate of dehiscence or infection between the groups. PMID- 16706764 TI - Frequency of use of suturing and repair techniques preferred by dermatologic surgeons. AB - BACKGROUND: There are many closure techniques and suture types available to cutaneous surgeons. Evidence-based data are not available regarding the frequency of use of these techniques by experienced practitioners. OBJECTIVE: To quantify, by anatomic site, the frequency of use of common closure techniques and suture types by cutaneous surgeons. METHOD: A prospective survey of the members of the Association of Academic Dermatologic Surgeons that used length-calibrated visual analog scales to elicit the frequency of use of specific suture techniques. RESULTS: A response rate of 60% (61/101) indicated reliability of the received data. Epidermal layers were closed most often, in descending order, by simple interrupted sutures (38-50%), simple running sutures (37-42%), and vertical mattress sutures (3-8%), with subcuticular sutures used more often on the trunk and extremities (28%). The most commonly used superficial sutures were nylon (51%) and polypropylene (44%), and the most common absorbable suture was polyglactin 910 (73%). Bilayered closures, undermining, and electrocoagulation were used, on average, in 90% or more sutured repairs. The median diameters (defined as longest extent along any axis) of most final wound defects were 1.1 to 2.0 cm (56%) or 2.1 to 3.0 cm (37%). Fifty-four percent of wounds were repaired by primary closure, 20% with local flaps, and 10% with skin grafting, with the remaining 15% left to heal by second intent (10%) or referred for repair (5%). Experience-related differences were detected in defect size and closure technique: defects less than 2 cm in diameter were seen by less experienced surgeons, and defects greater than 2 cm by more experienced surgeons (Wilcoxon's rank-sum test: p=.02). But more experienced surgeons were less likely to use bilayered closures (r=-0.28, p=.036) and undermining (r=-0.28, p=.035). CONCLUSIONS: There is widespread consensus among cutaneous surgeons regarding optimal suture selection and closure technique by anatomic location. More experienced surgeons tend to repair larger defects but, possibly because of their increased confidence and skill, rely on less complicated repairs. PMID- 16706765 TI - Mohs micrographic surgery for lentigo maligna and lentigo maligna melanoma using Mel-5 immunostaining: University of Minnesota experience. AB - BACKGROUND: Mohs micrographic surgery (MMS) continues to become a more common and accepted treatment for lentigo maligna (LM) and lentigo maligna melanoma (LMM). The primary difficulty encountered lies in the accurate identification of atypical single melanocytes to determine tumor-free margins. Numerous methods have been used to better visualize single melanocytes, with varying results. We present our experience using Mel-5 immunostaining in MMS of LM and LMM. METHODS: Two hundred patients with primary or recurrent LM or LMM were treated using MMS from 1999 to 2003 at the University of Minnesota. The initial clinical margins were determined by Wood's light examination, and an initial debulk specimen was taken and sent for formalin fixation and later reviewed by a dermatopathologist. The first Mohs layer was then taken, and staining with hemotoxylin and eosin as well as Mel-5 immunostaining was performed. All patients were followed up to evaluate for recurrence, with a mean follow-up time of 38.4 months. RESULTS: Of the 200 patients treated, only one recurrence was noted. This patient had been treated with excision followed by radiation before MMS. Use of Mel-5 immunostaining added approximately 40 minutes to each stage. Use of the Autostainer Immunostaining System (DAKO, Carpenterina, CA, USA) shortened the added time to 20 minutes. CONCLUSIONS: MMS with Mel-5 immunostaining yielded excellent results in the treatment of LM and LMM, with only one recurrence noted in 200 patients. When an automated immunostainer was used, minimal time was added to each Mohs stage. PMID- 16706766 TI - Facial lipoatrophy and the place of ultrasound. AB - BACKGROUND: Interest in facial lipoatrophy (LA) has recently intensified; this phenomenon is linked to the rise in the number of people adversely affected by the condition as a side effect of antiretroviral treatment for HIV, combined with the growing number of cosmetic products that claim to be able to correct the appearance of LA. Despite growing awareness of the problem, there is at present no standard and accepted technique with which to assess the severity of LA. OBJECTIVE: This review explores facial LA, the use of ultrasound in the evaluation of facial LA, its advantages and disadvantages, and will place the technique in the context of other means of assessing regional skin and fat thickness. METHOD: Review of literature published on PubMed. RESULTS Ultrasound, as with any technique used to assess facial LA, is associated with distinct advantages and disadvantages. CONCLUSIONS: Studies that use a number of different techniques to evaluate changes in dermal thickness provide the greatest insight into both perceived and actual changes in facial LA. Further investigation into the use of these techniques is warranted, along with a formal consensus of facial LA grades. PMID- 16706767 TI - ASDS guidelines of care for tumescent liposuction. PMID- 16706768 TI - Combination forceps fuse both safety and efficiency. AB - BACKGROUND: Instrumentation prevents needle stick injury. OBJECTIVE: To review forceps that insure safety and facilitate tissue-handling and knot-tying efficiency. METHOD: Medical literature reports were reviewed using Ovid. Commercially available instruments were qualitatively tested. RESULTS: Suture platforms securely hold suture needles and can be used during knot tying. A wide range of combination forceps have been invented and can be broadly categorized as either skin hook or toothed combination forceps. CONCLUSIONS: Combination forceps fuse both efficiency and safety. Skin hook forceps may eventually be the optimal combination instrument, but toothed combination forceps are recommended. PMID- 16706769 TI - Bilateral vermilion rotation flap. AB - BACKGROUND: Principles of aesthetic reconstruction recommend repair within cosmetic units with tissue of similar color, texture, and thickness. Because of the central facial location and primary functional and aesthetic importance of the lips, successful repair is critical in this location. Numerous procedures to reconstruct the lip have been reported; however, many result in a significant decrease in the oral aperture as well as dysesthesias, limitation of mobility, and significant scarring. OBJECTIVE: The purpose of this study was to describe a reconstructive option that repairs with skin of similar characteristics and avoids crossing into adjacent cosmetic subunits or sacrifice of normal skin or muscle. The proposed procedure is able to repair surgical defects up to 40% of the vermilion width in one stage and minimize the risk of nerve damage, scarring, or functional impairment. METHODS: The bilateral vermilion rotation flap permits repair within cosmetic subunits, repairs with skin of identical surface characteristics, and avoids surgery to the adjacent cutaneous or muscular portions of the lip. By keeping the pedicle intact, injury to neurovascular structures is minimized. RESULTS: The procedure provides an excellent repair for selected defects of the vermilion lip by following guidelines for optimum aesthetic and functional repair (i.e., repair within cosmetic subunits using skin of identical characteristics while avoiding trauma to the uninvolved cutaneous lip or orbicularis oris). CONCLUSION: The bilateral vermilion rotation flap is a useful technique for repair of surgical defects on the vermilion lip. It avoids unnecessary sacrifice of cutaneous or muscular lip tissue and provides an excellent cosmetic and functional result. PMID- 16706770 TI - Shark island pedicle flap for repair of combined nasal ala-perialar defects. AB - BACKGROUND: The combined nasal ala-perialar defect involving the concave intersection of the lateral nasal ala, nasal sidewall, cheek, and upper cutaneous lip is a problem for reconstructive surgery. During repair of combined cheek and nose defects, it is important not to blunt the alar facial sulcus. Defects involving these adjacent cosmetic units can be repaired by using combination procedures such as a flap/graft. OBJECTIVE: Our purpose is to introduce, describe, and illustrate a one-stage flap repair descriptively named the "shark" island pedicle flap (SIPF). The SIPF was developed for a specific combined nasal ala-perialar defect. This reconstruction restores the natural contours, preserves cosmetic boundaries, and eliminates the need for pexing sutures and graft/flap combinations. METHODS: The SIPF is an island pedicle flap with a superior arm that rotates 90 degrees into the wound. This arm repairs the alar portion of the defect. The advancing island pedicle flap repairs the alar facial sulcus. The 90 degrees rotation of the superior arm forces the alar portion of the flap to tilt 90 degrees relative to the remaining body of the flap, forming an inverted cone of redundancy. Natural re-creation of the lateral ala and the alar facial sulcus results. Illustrative examples with a descriptive technique are provided for the SIPF. RESULTS: A well-planned SIPF reconstruction can provide exceptional cosmetic and functional results. CONCLUSION: Cutaneous reconstructive surgeons will find the SIPF useful and reproducible in their armamentarium for single stage aesthetic and functional repair of a specific combined lateral ala-adjacent perialar tissue defect. PMID- 16706771 TI - Inverted island pedicle transposition for closure of full-thickness nasal mucosa defects. PMID- 16706773 TI - Level of confidence in diagnosis: clinical examination versus dermoscopy examination. AB - BACKGROUND: Confidence is an important factor in decision making and may influence patient care. OBJECTIVES: To evaluate whether short-training-based dermoscopy increases confidence in the diagnosis of skin lesions. METHODS AND MATERIALS: After a 1-hour course on dermoscopy, 20 pairs of clinical and dermoscopic images of lesions were presented to 19 dermatology residents with little or no dermoscopy experience. After viewing the clinical image, they were asked to assess their confidence in the diagnosis in a seven-point scale, with 1 reflecting that the respondent was 100% confident that the lesion was benign, while number 7 reflected 100% confidence that it was malignant. The same technique was used for dermoscopic images. RESULTS: Ten of the 20 pairs of evaluations showed a significant difference (p<.05). The largest differences were observed in lesions where clinical scores suggested that participants were uncertain about the diagnosis, but tended to decide that the lesion was benign after dermoscopy. Dermoscopy did not improve confidence in the evaluation of dysplastic lesions as well as lesions with obvious clinical diagnoses. CONCLUSIONS: Short-training-based dermoscopy improved confidence in the diagnosis of clinically challenging skin lesions, but the impact was not demonstrable for clinically obvious lesions and dysplastic nevi. PMID- 16706772 TI - Melanoacanthoma simulating pigmented spitz nevus: an unusual dermoscopy pitfall. AB - BACKGROUND: The starburst pattern is the dermoscopic hallmark of pigmented Spitz nevus, although it has been rarely observed in melanoma as well. OBJECTIVE: To describe a case of melanoacanthoma simulating pigmented Spitz nevus. MATERIAL AND METHODS: Clinical, dermoscopic, and histopathologic examinations were performed for the occurrence of a 4-mm pigmented skin lesion on the hip of a 38-year-old Caucasian woman. RESULTS: Dermoscopy examination of the lesion disclosed a stereotypical starburst pattern characterized by pigmented streaks symmetrically distributed at the periphery. A preoperative diagnosis of pigmented Spitz nevus was made, and the lesion was excised. However, subsequent histopathologic examination revealed a melanoacanthoma. CONCLUSION: The starburst pattern, although diagnostic for pigmented Spitz nevus, can be rarely observed in other benign or malignant pigmented skin lesions. Accordingly, all lesions in adults exhibiting a starburst pattern or other spitzoid features should be excised for histopathologic evaluation. PMID- 16706774 TI - Treatment of unilateral compensatory sweating after endoscopical thoracic sympathectomy for general hyperhidrosis with botulinum toxin A. PMID- 16706775 TI - Mixed skin ulcers misdiagnosed as pyoderma gangrenosum and rheumatoid ulcer: successful treatment with ultrasound-guided injection of polidocanol microfoam. PMID- 16706776 TI - Giant primary cutis verticis gyrata successfully treated with surgical excision and tissue expansion. PMID- 16706777 TI - Superficial squamous cell carcinomas and extensive actinic keratoses of the scalp treated with radiation therapy. PMID- 16706778 TI - Disseminated intravascular coagulation unmasked by Mohs micrographic surgery. PMID- 16706779 TI - Use of biologic dressing as a temporary wound dressing in reconstruction of a significant forehead Mohs defect. PMID- 16706780 TI - Suture granuloma following surgical neck rejuvenation procedure. PMID- 16706781 TI - Utility of hair clips for scalp surgeries. PMID- 16706782 TI - Increase in surgical procedures in a dermatology department in United Kingdom. PMID- 16706783 TI - New insight into the diagnosis of fastidious bacterial endocarditis. AB - Sterile blood cultures are noted in one third of patients with infectious endocarditis. Although in half of cases this is due to previous antibiotic therapy, in the other half, the aetiology of culture-negative endocarditis is intracellular bacteria such as Coxiella burnetii or fastidious growing bacteria. Although it was previously considered that the prevalence of such organisms was identical throughout the world, recent investigations on Bartonella endocarditis clearly showed that the aetiology of culture-negative endocarditis is likely to be strongly related to epidemiology of the agent in each country. During the past decade the use of molecular techniques such as PCR with subsequent sequencing to detect or to identify bacteria in valves from patients with infectious endocarditis have considerably improved the aetiological diagnosis. This is especially true in the case of culture-negative endocarditis following earlier antibiotic therapy. However, the fact that DNA remnants of past endocarditis can be detected some time after the acute episode, when the patient has been cured, suggests that the predictive value of these techniques along with the traditional histology and culture need to be evaluated closely. PMID- 16706784 TI - Recent developments in the immunology and biology of Malassezia species. AB - Malassezia spp. are members of the normal cutaneous flora, but are also associated with several cutaneous diseases. Recent studies of the interaction of Malassezia spp. with melanocytes, fibroblasts, keratinocytes and dendritic cells have highlighted their potential to modulate the immune response directed against them. In normal skin they may downregulate the inflammatory response, allowing them to live as commensals. In contrast, in atopic/eczema dermatitis syndrome and psoriasis, they may elicit an inflammatory response that contributes to the maintenance of lesions. Future research may define ways to influence this inflammatory cycle and hence to control or prevent exacerbations of these diseases. PMID- 16706785 TI - Recent advances in the immunology and diagnosis of echinococcosis. AB - Echinococcosis is a cosmopolitan zoonosis caused by adult or larval stages of cestodes belonging to the genus Echinococcus (family Taeniidae). The two major species of medical and public health importance are Echinococcus granulosus and Echinococcus multilocularis, which cause cystic echinococcosis and alveolar echinococcosis, respectively. Both cystic echinococcosis and alveolar echinococcosis are serious diseases, the latter especially so, with a high fatality rate and poor prognosis if managed inappropriately. This review highlights recent advances in immunity to infection and vaccination against both parasites in their intermediate and definitive hosts and procedures for diagnosis of cystic echinococcosis and alveolar echinococcosis, including the value of immunodiagnostic and DNA approaches. There is discussion also of progress in genomics and related technologies that is providing valuable insights on the functional biology of the Echinococcus organisms. These studies will underpin future research that will reveal a better understanding of the Echinococcus-host interplay, and suggest new avenues for the identification of additional targets for diagnosis, vaccination and chemotherapy. PMID- 16706786 TI - Legionella pneumophila in commercial bottled mineral water. AB - Sixty-eight commercial bottled mineral waters (64 brands, 68 different 'best before dates') were tested for the presence of bacteria and fungi. Six samples were Legionella antigen positive and six were Legionella pneumophila PCR positive. Two samples were both Legionella antigen and L. pneumophila PCR positive. Legionella cultures were negative. Although the PCR might have detected only dead Legionella cells, the PCR has been described to detect specifically viable but not culturable (VBNC) L. pneumophila cells as well. Whether VBNC bacteria may be present in bottled mineral waters and the risk for infection this may pose for severely immunocompromised patients should be investigated. PMID- 16706787 TI - T-h-2 immunity and CD3+CD45RBlow-activated T cells in mice immunized with recombinant bacillus Calmette-Guerin expressing HIV-1 principal neutralizing determinant epitope. AB - The genetic engineering of Mycobacterium bovis-bacillus Calmette-Guerin to express foreign epitopes is an attractive strategy in the field of epitope vaccines. We constructed an 'epitope-trap vector' with Mycobacterium tuberculosis chaperonin-10 as a carrier antigen and used it to express the HIV-1 principal neutralizing determinant epitope. We also identified a new chaperonin-10 promoter that was hyperexpressive compared with the heat shock protein-65 promoter. Splenocytes from recombinant bacillus Calmette-Guerin-immunized mice showed enhanced lymphocyte proliferation and interleukin-4 (but not interferon-gamma) secretion. The recombinant bacillus Calmette-Guerin-immunized group also exhibited mild delayed-type hypersensitivity reaction and a high frequency of CD3+CD45RBlow-activated T cells, together with high titer of antiprincipal neutralizing determinant immunoglobulin G antibodies. Thus, this epitope delivery system induced strong epitope-specific T-h-2 polarization. PMID- 16706789 TI - Characterization of the effects of butyric acid on cell proliferation, cell cycle distribution and apoptosis. AB - We examined concentration-dependent changes in cell cycle distribution and cell cycle-related proteins induced by butyric acid. Butyric acid enhanced or suppressed the proliferation of Jurkat human T lymphocytes depending on concentration. A low concentration of butyric acid induced a massive increase in the number of cells in S and G2/M phases, whereas a high concentration significantly increased the accumulation of cells in G2/M phase, suppressed the accumulation of cells in G0/G1 and S phases, and induced apoptosis that cell cycle-related protein expression in Jurkat cells treated with high levels of butyric acid caused a marked decrease in cyclin A, cyclin E, cyclin-dependent kinase 2 (CDK2), CDK4 and CDK6 protein levels in G0/G1 and S phases, with apoptosis induction, and a decrease in cyclin B, Cdc25c and p27KIP1 protein levels, as well as an increase in p21CIP1/WAF1 protein level, in the G2/M phase. Taken together, our results indicate that butyric acid has bimodal effects on cell proliferation and survival. The inhibition of cell growth followed by the increase in apoptosis induced by high levels of butyric acid were related to an increase in cell death in G0/G1 and S phases, as well as G2/M arrest of cells. Finally, these results were further substantiated by the expression profile of butyric acid-treated Jurkat cells obtained by means of cDNA array. PMID- 16706788 TI - Importance of complement 3 and mannose receptors in phagocytosis of Paracoccidioides brasiliensis conidia by Nramp1 congenic macrophages lines. AB - Genetic factors influence susceptibility to Paracoccidioidomycosis, a Latin American endemic mycosis. The pattern of susceptibility of congenic mouse strains infected with Paracoccidioides brasiliensis resembles the pattern of the Nramp1 gene. Thus, congenic murine bone-marrow-derived macrophage lines B10R (Nramp1rGly169) and B10S (null Nramp1 protein expression, Nramp1sAsp169) were infected with P. brasiliensis conidia and compared, under opsonic and nonopsonic conditions. Opsonization increased the percentage of phagocytosis by both cell lines. B10R macrophages exhibited a higher percentage of cells with associated conidia and higher number of conidia per macrophage than B10S. Heat-inactivation and EDTA treatment of serum used for opsonization, and treatment of macrophages with anti-complement receptor 3 (CR3) decreased phagocytosis by both cell lines. alpha-methyl-d-mannoside reduced phagocytosis by B10R macrophages, suggesting that the mannose receptor participates in phagocytosis by these cells. The CR3 expression was similar on both cell lines and B10R expressed more mannose receptors, but neither cell line expressed CR1. IFNgamma decreased the conversion of conidia to the yeast form of P. brasiliensis in B10R, but not in B10S macrophages. PMID- 16706790 TI - Carbohydrate-containing components of biofilms produced in vitro by some staphylococcal strains related to orthopaedic prosthesis infections. AB - The capacity of coagulase-negative staphylococci to colonize implanted medical devices is generally attributed to their ability to produce biofilms. Biofilm of the model strain of Staphylococcus epidermidis RP62A was shown to contain two carbohydrate-containing moieties, a linear poly-beta-(1-->6)-N-acetyl-D glucosamine (PNAG) and teichoic acid. In the present study, we investigated several biofilm-producing staphylococci isolated from infected orthopaedic implants and characterized the composition of the laboratory-grown biofilms using chemical analysis and 1H nuclear magnetic resonance spectroscopy. Extracellular teichoic acid was produced by all strains studied. Some of the clinical strains were shown to produce biofilms with compositions similar to that of the model strain, containing a varying amount of PNAG. The chemical structure of PNAG of the clinical strains was similar to that previously described for the model strains S. epidermidis RP62A and Staphylococcus aureus MN8m, differing only in the amount of charged groups. Biofilms of the strains producing a substantial amount of PNAG were detached by dispersin B, a PNAG-degrading enzyme, while being unsusceptible to proteinase K treatment. On the other hand, some strains produced biofilms without any detectable amount of PNAG. The biofilms of these strains were dispersed by proteinase K, but not by dispersin B. PMID- 16706791 TI - Balamuthia mandrillaris exhibits metalloprotease activities. AB - Balamuthia mandrillaris is a recently identified protozoan pathogen that can cause fatal granulomatous encephalitis. However, the pathogenesis and pathophysiology of B. mandrillaris encephalitis remain unclear. Because proteases may play a role in the central nervous system (CNS) pathology, we used spectrophotometric, cytopathic and zymographic assays to assess protease activities of B. mandrillaris. Using two clinical isolates of B. mandrillaris (from human and baboon), we observed that B. mandrillaris exhibits protease activities. Zymographic assays revealed major protease bands of approximate molecular weights in the region of 40-50 kDa on sodium dodecyl sulfate polyacrylamide gels using gelatin as substrate. The protease bands were inhibited with 1,10-phenanthroline, suggesting metallo-type proteases. The proteolytic activities were observed over a pH range of 5-11 with maximum activity at neutral pH and at 42 degrees C. Balamuthia mandrillaris proteases exhibit properties to degrade extracellular matrix (ECM), which provide structural and functional support to the brain tissue. This is shown by degradation of collagen I and III (major components of collagenous ECM), elastin (elastic fibrils of ECM), plasminogen (involved in proteolytic degradation of ECM), as well as other substrates such as casein and gelatin but not haemoglobin. However, these proteases exhibited a minimal role in B. mandrillaris-mediated host cell death in vitro using human brain microvascular endothelial cells (HBMECs). This was shown using broad-spectrum matrix metalloprotease inhibitors, GM 6001 and GM 1489, which had no effect on B. mandrillaris-mediated HBMEC cytotoxicity. This is the first demonstration that B. mandrillaris exhibits metalloproteases, which may play important role(s) in the ECM degradation and thus in CNS pathology. PMID- 16706792 TI - Proinflammatory cytokine and chemokine modulation by Streptococcus suis in a whole-blood culture system. AB - Streptococcus suis is an important swine and human pathogen. Inflammation, a hallmark of S. suis infection, is thought to be responsible for most clinical signs of meningitis, septicaemia and sudden death. In this work, using a porcine whole blood model, S. suis serotype 2 was shown to trigger the release of several pro-inflammatory cytokines as evaluated by reverse transcriptase-PCR and enzyme linked immunosorbent assay. Although individual variations were observed among different S. suis strains, no correlations were observed between the strain origin/phenotype and cytokine levels. Live bacteria induced higher tumour necrosis factor alpha, interleukin-1 beta (IL-1beta) and IL-6 levels than did heat-killed bacteria. In contrast, heat-killed bacteria stimulated higher levels of IL-8 and monocyte chemotactic protein one (MCP-1). The bacterial cell wall was observed to be the major cytokine-inducting components, whereas capsule expression was important for MCP-1 activation. The presence of specific antibodies suppressed bacterial growth resulting in significantly reduced levels of cytokine production. Thus, antibody-mediated bacterial phagocytosis combined with suppressed inflammation may be beneficial for infection control strategies. We provide first evidence of S.suis-induction of pro-inflammatory swine cytokines and demonstrate the strength and relevance of the whole blood culture systems in the investigation of S. suis modulation of cytokine production. PMID- 16706793 TI - Intratracheal immunization with pili protein protects against mortality associated with Pseudomonas aeruginosa pneumonia in mice. AB - We examined the protective effect of intratracheal immunization with Pseudomonas aeruginosa pili protein against respiratory infection caused by P. aeruginosa. Mice were immunized intratracheally or subcutaneously with purified pili protein or bovine serum albumin as a control. Intratracheally but not subcutaneously pili protein-immunized mice showed significant improvement of survival after intratracheal challenge with the PAO1 strain. Furthermore, bacterial cell counts in pili protein-immunized murine lungs were significantly decreased compared to controls at 18 h after the challenge. Antipili protein antibody titers in bronchoalveolar lavage fluid of intratracheally pili protein-immunized mice were higher than in bovine serum albumin immunized mice. However, antipili antibody titers were not increased in bronchoalveolar lavage fluid of subcutaneously pili protein-immunized mice, despite the high serum antipili antibody titers. Inoculation of P. aeruginosa induced immediate increases in interleukin-12 and interferon-gamma in bronchoalveolar lavage fluid of pili protein-immunized mice, reflecting an adequate and rapid immune response against P. aeruginosa respiratory tract infection. Our findings suggest that intratracheal pili protein immunization is effective against respiratory tract infection caused by P. aeruginosa in mice. PMID- 16706795 TI - Rapid detection and clinical features of infants and young children with acute lower respiratory tract infection due to respiratory syncytial virus. AB - During December to the end of February of 2003 and 2004, a total of 282 nasopharyngeal aspirates were obtained from infants and young children admitted to the Buraidah Maternity and Pediatric Hospital, Al-Qassim, Saudi Arabia, and clinically diagnosed as suffering from acute lower respiratory tract infections. The aspirates were tested for the presence of respiratory syncytial virus using direct fluorescein-labeled monoclonal antibody assay. Of the 282 specimens, 128 (45.4%) were found to be positive for respiratory syncytial virus. The most positive specimens came from patients less than one year old (51.3%), and were associated with bronchopneumonia (56.7%) or bronchiolits (55.4%). Coughing (100%) and tachpnea (98%) were significantly more frequent in infants with respiratory syncytial virus infection, followed by wheezing, crepitation and retraction, each representing 66%. Three deaths were reported. The availability of a rapid viral diagnostic assay will be an important tool for physicians to make more accurate treatment decisions and therefore reduce unnecessary antibiotic usage and hospital stay for the patients. PMID- 16706794 TI - Length of the linker and the interval between immunizations influences the efficacy of Vibrio cholerae O1, Ogawa hexasaccharide neoglycoconjugates. AB - Ogawa hexasaccharide neoglycoconjugates induce protective antibodies in mice. Similar Ogawa conjugates but with a longer linker that connects the carrier to shorter saccharides are immunogenic, but generally ineffective at inducing vibriocidal or protective antibodies. The efficacy of Ogawa hexasaccharide neoglycoconjugates of different linker lengths were tested. The majority of mice given immunizations separated by a 14-day gap did not produce vibriocidal or protective antibodies. Mice immunized 28 days apart with immunogens containing the shortest or medium length linker, but not the longest, produced vibriocidal and protective antibodies. A nonprotective, priming dose of purified Ogawa LPS followed 5 days later with a booster of the Ogawa neoglycoconjugates (di-, tetra , or hexasaccharide) resulted in vibriocidal antibodies at day 10. PMID- 16706796 TI - A novel erythrocyte-based immunoassay for simultaneous detection of both antimycobacterial antibody response and mycobacterial antigen in human serum samples of pulmonary tuberculosis and a control group of patients using 'a single probe'. AB - A modified passive hemagglutination using double aldehyde stabilized cells (tanned sheep erythrocytes treated with glutaraldehyde and pyruvic aldehyde) was evaluated for detection of both antimycobacterial antibodies and circulating mycobacterial antigens simultaneously in human serum samples from patients with pulmonary tuberculosis (n=40) and a control group (n=44). Double aldehyde stabilized cells sensitized with an optimum dose of 200 microg mL(-1) of sonicate extract of Mycobacterium tuberculosis antigens was used as single probe to detect both antibodies and antigen, respectively, by passive hemagglutination and passive hemagglutination inhibition. The sensitivity limit of passive hemagglutination inhibition was determined to be 280 ng mL(-1) using a dose response curve. Sensitivity of passive hemagglutination and passive hemagglutination inhibition, respectively, was 90% and 52.5%, and specificity was 91% and 100%. Although passive hemagglutination and passive hemagglutination inhibition need further evaluation, these erythrocyte-based immunoassays are potentially advantageous, especially as double aldehyde stabilized sensitized cells could be used as a single probe for detection of both antibodies and antigen. In addition, erythrocyte-based immunoassays are rapid, simple and cost effective with a high degree of sensitivity. PMID- 16706798 TI - Limited contribution of Toll-like receptor 2 and 4 to the host response to a fungal infectious pathogen, Cryptococcus neoformans. AB - The present study was designed to elucidate the role of Toll-like receptor (TLR) 2 and TLR4 in the host response to Cryptococcus neoformans. Both TLR2 knockout (KO) and TLR4KO mice produced interleukin-1beta (IL-1beta), IL-6, IL-12p40 and tumor necrosis factor-alpha (TNF-alpha) in sera and cleared this fungal pathogen from infected lungs at a comparable level to control littermate (LM) mice. Synthesis of these cytokines was not significantly different in the lungs of these KO mice and LM mice, although IL-1beta, IL-6 and IL-12p40 tended to be lower in TLR2KO, but not TLR4KO, mice than in controls. In addition, there was no significant reduction detected in the synthesis of IL-12 and TNF-alpha by bone marrow-derived dendritic cells from TLR2KO and TLR4KO mice upon stimulation with live yeast cells. Finally, HEK293 cells expressing either TLR2/dectin-1 or TLR4/MD2/CD14 did not respond to C. neoformans in the activation of nuclear factor kappa B (NFkappaB) detected by a luciferase assay. Our results suggest that TLR2 and TLR4 do not or only marginally contribute to the host and cellular response to this pathogen. PMID- 16706797 TI - Role of SLC11A1 (formerly NRAMP1) in regulation of signal transduction induced by Toll-like receptor 7 ligands. AB - Modulation of immune responses using Toll-like receptor (TLR) ligands is fast becoming one of the main new approaches for the treatment of infectious and allergic diseases. Characterizing the role of genetic factors in modulating responses to these ligands will be crucial in determining the efficacy of a particular treatment. Our previous findings have shown that treatment of Mycobacterium bovis BCG infection with a synthetic TLR7 ligand resulted in a reduction of the splenic bacterial load only in mice carrying a wild-type allele of Nramp1. To understand further how natural resistance-associated macrophage protein 1 (NRAMP1) modulates responses to TLR7 ligands, we have analysed various important TLR7 signal transduction events in macrophage cell lines derived from B10.ANramp1r and B10.ANramp1-/- mice. The Nramp1 genotype did not affect TLR7 receptor expression, ligand uptake or intracellular processing. Following TLR7 ligand stimulation, p38 mitogen-activated protein kinase (MAPK) activation was significantly reduced in B10A.Nramp1-/- macrophages compared with B10A.Nramp1r cells. Interestingly, levels of protein kinase C zeta (PKCzeta) activation were also found to be lower in B10A.Nramp1-/- macrophages and inhibition of this kinase in B10A.Nramp1r cells led to a reduction in cytokine production. Taken together, the data demonstrate a role for NRAMP1 in modulating p38 MAPK and PKCzeta activity, which leads to reduced cytokine induction by TLR7 ligands. PMID- 16706799 TI - The maturation of the Journal of Gastroenterology and Hepatology ( JGH) and the JGH Foundation. PMID- 16706800 TI - Journal of Gastroenterology and Hepatology: 20 years and gaining momentum. PMID- 16706801 TI - Hepatocellular carcinoma and non-alcoholic fatty liver disease: Major liver diseases emerging from the Asia-Pacific Region. PMID- 16706802 TI - History of the Journal of Gastroenterology and Hepatology. PMID- 16706803 TI - 'Yes, now you are free from chronic HCV infection, but . . .'. PMID- 16706804 TI - Helicobacter pylori virulence: does it matter in patients with non-ulcer dyspepsia? PMID- 16706805 TI - Treating NASH. PMID- 16706806 TI - Review of genetic and epigenetic alterations in hepatocarcinogenesis. AB - Hepatocellular carcinoma (HCC) is associated with multiple risk factors and is believed to arise from pre-neoplastic lesions, usually in the background of cirrhosis. However, the genetic and epigenetic events of hepatocarcinogenesis are relatively poorly understood. HCC display gross genomic alterations, including chromosomal instability (CIN), CpG island methylation, DNA rearrangements associated with hepatitis B virus (HBV) DNA integration, DNA hypomethylation and, to a lesser degree, microsatellite instability. Various studies have reported CIN at chromosomal regions, 1p, 4q, 5q, 6q, 8p, 10q, 11p, 16p, 16q, 17p and 22q. Frequent promoter hypermethylation and subsequent loss of protein expression has also been demonstrated in HCC at tumor suppressor gene (TSG), p16, p14, p15, SOCS1, RIZ1, E-cadherin and 14-3-3 sigma. An interesting observation emerging from these studies is the presence of a methylator phenotype in hepatocarcinogenesis, although it does not seem advantageous to have high levels of microsatellite instability. Methylation also appears to be an early event, suggesting that this may precede cirrhosis. However, these genes have been studied in isolation and global studies of methylator phenotype are required to assess the significance of epigenetic silencing in hepatocarcinogenesis. Based on previous data there are obvious fundamental differences in the mechanisms of hepatic carcinogenesis, with at least two distinct mechanisms of malignant transformation in the liver, related to CIN and CpG island methylation. The reason for these differences and the relative importance of these mechanisms are not clear but likely relate to the etiopathogenesis of HCC. Defining these broad mechanisms is a necessary prelude to determine the timing of events in malignant transformation of the liver and to investigate the role of known risk factors for HCC. PMID- 16706807 TI - Enteral versus parenteral nutritional support in acute pancreatitis: a clinical review. AB - A critical analysis of the trials comparing enteral with parenteral nutritional support in acute pancreatitis is presented with particular scrutiny of the clinical endpoints in each trial. The heterogeneity of the trials and the discrepancies in caloric intake between enterally and parenterally fed patients are discussed. The hazards of each route of nutritional support are highlighted and the concept of 'nutritional bioavailability' in the context of acute pancreatitis is introduced. It is concluded that although there is evidence to support enteral feeding as the preferred option, both routes have a role. PMID- 16706808 TI - Biliary excretion of sulfated bile acids and organic anions in zone 1- and zone 3 injured rats. AB - BACKGROUND AND AIMS: There are several reports on the biliary excretion of bile acids and organic anions in zone 1- and zone 3-injured rat liver, but the results are controversial. In order to dissolve the discrepancy between previous works about the role of hepatic zonation on the hepatic handling of the substrates of multidrug resistance protein 2, the biliary excretion of sulfated bile acids, pravastatin and phenolphthalein glucuronide was studied in zone 1- and zone 3 injured rats. METHODS: Zone 1 and zone 3 injury were caused by allyl alcohol and bromobenzene, respectively. Bile acid sulfates, pravastatin and phenolphthalein glucuronide were administered i.v. to bile duct-cannulated rats, and their biliary excretion was studied. RESULTS: The biliary excretion of a tracer dose of taurolithocholate-3-sulfate and its excretory maximum were unchanged in zone 1 injury, but were diminished in zone 3 injury, whereas the biliary excretion of taurochenodeoxycholate-3-sulfate was unchanged in zone 1 and zone 3 injury. The biliary excretion of pravastatin and phenolphthalein glucuronide was markedly decreased only in zone 3 injury, whereas the excretory maximum of phenolphthalein glucuronide was decreased in both zone 1 and zone 3 injury. CONCLUSIONS: These findings indicate that zone 3 is important for the biliary excretion of substrates of multidrug resistance protein 2. PMID- 16706809 TI - gammadelta T cells increase with gastric mucosal interleukin (IL)-7, IL-1beta, and Helicobacter pylori urease specific immunoglobulin levels via CCR2 upregulation in Helicobacter pylori gastritis. AB - BACKGROUND AND AIMS: The purpose of this study was to investigate possible factors that could impact on gammadelta T cell accumulation in the gastric mucosa. METHOD: Subjects were 22 Helicobacter pylori (H. pylori)-free and 75 H. pylori-infected mucosa biopsies classified into grades I approximately III gastritis as per our previous study. The number of gammadelta- and 45 RO-positive T cells were determined by immunostaining. Gastric mucosal anti-H. pylori urease specific antibodies and interleukin (IL)-1beta, IL-2, 4, 7, 10 and IL-12 levels were assayed by enzyme-linked immunosorbent assay (ELISA). CC chemokine receptor 2 (CCR2) expression levels, migration, and cytokine production in gammadelta T cells stimulated by H. pylori urease were also evaluated. RESULTS: The gammadelta T cell count was significantly higher in grade III gastritis which exhibits strong immunoglobulin (Ig)A and IgG responses to H. pylori urease with lymphoid follicles than in other groups. gammadelta T cell count was significantly correlated with IL-1beta and interleukin-7 (IL-7) levels in the gastric mucosa. H. pylori urease immunoreactivity was detected in lamina propria of grade III gastritis, along with many gammadelta T cells. After H. pylori eradication therapy, the gammadelta T cell count in grade III gastritis significantly decreased. H. pylori urease stimulated significant increases in CCR2 expression levels, although to a lesser degree than those induced by IL-7 stimulation in both peripheral and mucosal gammadelta T cells. Interferon (IFN)-gamma and IL-10 production was also stimulated by H. pylori urease in peripheral gammadelta T cells. CONCLUSIONS: Gastric mucosal increases in IL-7 and IL-1beta closely corresponded to the accumulation of gammadelta T cells in gastric mucosa. An association was also seen between gammadelta T cell accumulation and H. pylori urease-specific Ig levels. PMID- 16706811 TI - Short-term effects of magnetic sacral dermatome stimulation for idiopathic slow transit constipation: sham-controlled, cross-over pilot study. AB - BACKGROUND AND AIM: An increase in recto-sigmoid colon activity through electrical stimulation of the sacral dermatomes has previously been reported. It has not been evaluated whether or not sacral dermatome stimulation has beneficial effects on constipation symptoms and anorectal function in constipated patients. Our aim was to evaluate short-term effects of magnetic stimulation of the sacral dermatomes on constipation symptoms and anorectal function in patients with idiopathic slow transit constipation. METHOD: Fourteen patients with idiopathic slow transit constipation were enrolled. Constipation symptoms, stool form and anorectal function were assessed before treatment, and at 3 and 6 weeks of treatment. Six-week treatment consisted of either a 3-week period of sham treatment or a 3-week period of magnetic stimulation of the S2-S3 dermatomes, which was performed in a randomized cross-over design. RESULTS: During the stimulation period, the frequency score of spontaneous bowel movements decreased in eight of the 14 patients (2.9 [2-3]vs 1.4 [0-2]), whose threshold volumes for urge to defecate and maximum tolerable volumes were significantly greater than those of the non-responders, and significantly decreased at the end of treatment. The degree of straining on defecation also significantly decreased in the responders. Responders had shorter right colonic transit time and longer left colonic transit time compared to the non-responders. Sham treatment did not affect constipation symptoms, stool form and rectal sensation. CONCLUSION: Sacral dermatome stimulation may offer potential for therapeutic benefit for a subset of patients with idiopathic slow transit constipation, particularly constipated patients with rectal hyposensation or hindgut dysfunction. PMID- 16706810 TI - Attitudes and knowledge of colorectal cancer and screening in Hong Kong: a population-based study. AB - BACKGROUND: Colorectal cancer (CRC) ranks third of all cancer mortalities in Hong Kong. CRC screening can reduce mortality. However, participation in the CRC screening program depends on the knowledge and attitude of the population. The aim of this study is to investigate the knowledge and attitude towards CRC screening in the Hong Kong population. METHODS: One thousand and thirty-three patients were interviewed by telephone survey. Knowledge and attitudes were assessed by using questionnaires. RESULTS: Only 8.9% of the respondents were able to mention that CRC was the second commonest cancer cause of death; 4.5% believed they would have a higher chance of developing CRC as compared to other cancers, and 7.4% could mention one of the screening methods. Approximately half (50.3%) were unable to identify any available screening venue. Approximately one-third (29.5%) indicated that they would definitely join a free CRC screening program, but the percentage dropped to 7.9% when a paid screening program was offered. Some (30.4%) of the respondents had the wrong impression that they did not require CRC screening because they were asymptomatic. Multivariate analysis showed that the independent predictor for participating in free screening was the concern level of getting CRC, while that for a paid screening program was a positive family history of CRC and the concern level. The concern level was dependent on a positive family history and knowledge of CRC. CONCLUSION: The general population in Hong Kong had deficient knowledge of CRC and screening. Education on the impact of CRC and beneficial effects of CRC screening should be offered before launching a CRC screening program. PMID- 16706812 TI - Slow-wave photic stimulation relieves patient discomfort during esophagogastroduodenoscopy. AB - BACKGROUND AND AIM: Although pretreatment with a sedative drug is effective in relieving pain during esophagogastroduodenoscopy (EGD), such drugs can cause significant side-effects. The aim of this study was to examine the effect of slow wave photic stimulation on discomfort and/or pain felt during EGD. METHODS: Forty consecutive patients (25 men and 15 women) who underwent diagnostic EGD in our hospital were included in the study. Twenty patients received photic stimulation for 25 min, and underwent electroencephalographic recording, in addition to the usual premedications. Twenty control patients received the same treatment but without photic stimulation. All patients evaluated the discomfort/pain felt during endoscopy against a five-grade scale in comparison with what they had experienced in their previous examination. RESULTS: Patients with an improved discomfort/pain score were 18/20 and 3/20 in the treated and control groups, respectively. Overall comparison of pain scores between both groups was significant (P<0.0001). The proportion of slow-wave activity recorded in patients' electroencephalograms significantly increased in the treated group compared to control values (36.6+/-6.8% vs 29.1+/-3.4%, P<0.001). There was a close correlation between the degree of discomfort/pain felt during endoscopy and the proportion of slow-wave activity (P<0.001). CONCLUSION: Slow-wave photic stimulation shows promise as a treatment for relieving the discomfort and/or pain felt by patients undergoing EGD. PMID- 16706813 TI - Dietary influence on electro-gastrography and association of alterations in gastric myoelectrical activity with symptoms in patients with functional dyspepsia. AB - BACKGROUND AND AIM: It is unclear which type of meal is adequate in measuring electro-gastrography and which parameter of electro-gastrography is of clinical relevance. The aims of the present study were to compare the influence of water and a nutrient drink on electro-gastrography in patients with functional dyspepsia (FD) and in healthy volunteers, and to investigate the association of alterations of electro-gastrography with symptoms. METHOD: In 30 patients with FD and 12 healthy volunteers, the recording of electro-gastrography with symptom assessments was performed pre-prandially and postprandially. Subjects ingested the same volume of water and a nutrient drink at a fixed rate, which was performed in a randomized cross-over design. RESULTS: Unlike water, the power ratio after/before a nutrient drink was significantly lower in patients compared to controls. Patients had more severe bloating and epigastric pain after nutrient ingestion than after water intake. Absent postprandial increase of power was observed in seven of the 30 patients after nutrient intake, and in three of the 30 patients after water intake. The former patients had significantly more severe fullness and bloating. Irrespective of a test meal, the percentage rates of normogastria significantly decreased postprandially in the patient group, but not in the control group. No differences in symptom severity were observed between patients with and without abnormally low percentage rates of normogastria. CONCLUSIONS: Decreased postprandial rates of normogastria and absent postprandial increase of power are electro-gastrographic abnormalities found in a subset of patients with FD. A nutrient drink is more adequate for the detection of such alterations than water. The lack of a postprandial increase of power is associated with the severity of some dyspeptic symptoms, but decreased postprandial rates of normogastria are not. PMID- 16706814 TI - Colonic pseudolipomatosis, microscopically classified into two groups. AB - BACKGROUND: Colonic pseudolipomatosis is rare and the pathogenesis is controversial. The purpose of the present paper was to clarify endoscopic and histological characteristics of colonic pseudolipomatosis and to discuss the etiology. METHODS: A total of 15 lesions from 14 patients was reviewed. They were able to be histologically classified into two groups on the basis of variety in size of the vacuoles: Group A, the ratio of largest vacuole to smallest vacuole in size is less than three, Group B, the ratio is more than four. RESULTS: Four of 15 lesions were group A, and were endoscopically polypoid or flat lesions covered with normal-looking mucosa. They were microscopically characterized by (i) predominant location in the upper portion of the lamina propria; (ii) no submucosal involvement; (iii) less variation in vacuolar size; and (iv) no association with lymph follicles. The vacuoles of group A contained proteinaceous materials in two of four lesions. Group B (11 lesions) had small elevated mucosa with normal-looking surface or non-elevated reddish mucosa. Microscopically, the lesions were mainly located in the lower portion of the lamina propria, occasionally also in the submucosa, had variable-sized vacuoles, and were related to lymph follicles. CONCLUSION: It is suggested that the vacuoles in group A contain fluid, and may indicate an abnormal stagnation of interstitial fluid. Microscopic appearance of group B is essentially similar to that of pneumatosis coli. It is thought that group B probably results from penetration of gas from the crypts into the mucosa during colonoscopy. It is unclear why group B had a preference for ileocecal valve and an association with lymph follicles. PMID- 16706815 TI - Mucosal mast cell counts correlate with visceral hypersensitivity in patients with diarrhea predominant irritable bowel syndrome. AB - BACKGROUND AND AIM: Although increased mast cells in the gut and their role in visceral hypersensitivity in irritable bowel syndrome have been postulated, this relationship remains unclear. Our aim was to determine whether a relationship exists between the number of mucosal mast cells in the gut and visceral hypersensitivity. METHOD: Eighteen patients with diarrhea predominant irritable bowel syndrome (D-IBS) (eight females, 10 males aged 25-65 years; mean 42.6 years) with symptoms meeting the Rome-II criteria, and 15 healthy controls (five females, 10 males aged 31-57 years; mean 41.4 years) were recruited. Participants completed a questionnaire for bowel symptoms and psychological distress. Colonic mucosal mast cells were identified immunohistochemically and quantified by image analysis, and maximally tolerable pressures were evaluated using barostat test. RESULTS: Numbers of mast cells were significantly greater in the terminal ileum, ascending colon and rectum of D-IBS patients compared with controls (P<0.01). A multivariate analysis of the barostat test showed that maximally tolerable pressures of D-IBS patients were significantly lower than those of controls (P<0.01). When patients were divided into the rectal hypersensitivity (+) and (-) groups by the distension level of 34 mmHg, mast cell counts were significantly higher in the rectal hypersensitivity (-) group than in the rectal hypersensitivity (+) group for the terminal ileum, ascending colon and rectum (P<0.05, respectively). CONCLUSIONS: Rectal sensitivity was enhanced in D-IBS patients with moderately increased mucosal mast cells, but it was attenuated in patients with markedly increased ones. This study might provide evidence for an important role of mast cells in visceral hypersensitivity. PMID- 16706816 TI - Duodenal lymphangitis carcinomatosa: endoscopic characteristics and clinical significance. AB - BACKGROUND AND AIM: Duodenal lymphangitis carcinomatosa has been sporadically described, but so far little attention has been paid to duodenal lymphangitis carcinomatosa. METHODS: Four cases with duodenal lymphangitis carcinomatosa were endoscopically and histologically examined. RESULTS: The four cases exhibited multiple polypoid lesions along the Kerckring's folds and/or were covered by characteristically granular, non-ulcerated mucosa upon thickening. The granularity seems to been caused by dilated lymph vessels containing the carcinoma cells. The lesions were microscopically characterized by: (i) involvement of lymph vessels located in the upper portion of the lamina propria; (ii) no inflammatory changes; and (iii) no desmoplastic changes. Primary sites were thought to be the stomach in case 1, the pancreas in cases 2 and 4, and unknown in case 3. All patients died within 6 months after admission or endoscopic examination. CONCLUSIONS: As duodenal lymphangitis carcinomatosis shows characteristic endoscopic appearance, endoscopic diagnosis is not difficult. We should realize that the lesion represents extremely poor prognosis, and it should be distinguished from ordinary metastatic duodenal carcinoma. PMID- 16706817 TI - Fas-associated phosphatase-1 promotes Fas-mediated apoptosis in human colon cancer cells: novel function of FAP-1. AB - BACKGROUND AND AIM: Fas-associated phosphatase-1 (FAP-1) has been thought as an inhibitor in Fas-mediated apoptosis. Here, we investigated the role of FAP-1 in Fas-mediated apoptosis of human colon cancer cells. METHOD: The viability of four colon cancer cell lines treated with agonistic anti-Fas antibody was determined using WST-1 assay and cell death detection ELISA. pRc/CMV-FAP-1 was transfected to a FAP-1-negative, Fas-resistant colon cancer cell line SW480 by lipofection and the clones expressing FAP-1 protein were selected by limiting dilution. In the clones, expression of 550 genes was analyzed by cDNA microarrays. Protein expression of FAP-1 and molecules related to apoptosis was examined by western blot. RESULTS: We obtained two FAP-1 overexpressed clones which were much more susceptible to Fas-mediated apoptosis than control cells. In the clones, caspase 8 and caspase 3 were fully activated by agonistic anti-Fas antibody treatment. Bcl-2 family proteins were not related to the high susceptibility of these clones, because caspase 9 was not activated. Transfection of FAP-1 did not suppress the survival actions of insulin-like growth factor (IGF-1) which enhanced survival signal through Akt phosphorylation. Upregulation in 21 genes and downregulation in 29 genes was revealed by cDNA arrays. We confirmed protein expression of p21 and phosphorylated p21 were much more enhanced in the clones than in control cells. CONCLUSIONS: Overexpression of FAP-1 enhanced susceptibility to Fas-mediated apoptosis in SW480 and upregulation of p21 may contribute to this phenomenon. Our results indicate a novel function of FAP-1 in Fas-mediated apoptosis of human colon cancer cells. PMID- 16706818 TI - Polymorphism of CD14 gene but not the mutation of TLR4 gene is associated with colorectal cancer in Chinese patients. AB - OBJECTIVES: Toll-like receptor 4 and CD14 are components of the lipopolysaccharide receptor complex. Our study aimed to investigate an association between TLR4 Asp299Gly and CD14-260 polymorphisms in Chinese patients with colorectal cancer. METHOD: By a method of polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP), we genotyped TLR4 Asp299Gly and CD14-260 polymorphisms in 110 unrelated patients with colorectal cancer and 160 healthy controls from the Chinese Han population. RESULTS: We found significant differences in CD14 genotypes between healthy controls and patients with colorectal cancer. The frequency of the C/C genotype in healthy controls (15.6%) was significantly lower than in the group of colorectal cancer patients (32%). The frequency of the C/T genotype in healthy controls (48.1%) was significantly higher than that in the group of colorectal cancer patients (31%). No TLR4 Asp299Gly mutation was detected in any patients or healthy controls in the Chinese Han population. CONCLUSIONS: These findings indicated that the polymorphism of CD14 but not TLR4 Asp299Gly mutation was associated with Chinese patients with colorectal cancer, and the CD14 gene may contribute to the predisposition to colorectal cancer. Screening for the CD14 C-260T genotype is likely to be a useful tool for risk assessment or prognostication for colorectal cancer in the Chinese Han population. PMID- 16706819 TI - Endoscopic characteristics and Helicobacter pylori infection in NSAID-associated gastric ulcer. AB - BACKGROUND AND AIM: Helicobacter pylori infection and non-steroidal anti inflammatory drugs (NSAIDs) are deeply involved in the etiology of gastric ulcers. The aim of our study was to clarify the endoscopic characteristics and H. pylori infection status of NSAID-associated gastric ulcers. METHODS: The study group comprised 50 patients (23 men, 27 women; mean age 66.5 years) with NSAID associated gastric ulcers and 100 sex- and age-matched patients with gastric ulcer associated with other factors (control group). Ulcer morphology, size and number of lesions, onset site and incidence of hemorrhagic ulcers were investigated endoscopically in both groups. H. pylori infection was diagnosed by serology, histology and (13)C-urea breath test. RESULTS: Multiple lesions (68% vs 20%, P<0.001), occurrence in the antrum (56% vs 6%, P<0.001), and hemorrhagic ulcer (34% vs 4%, P<0.001) were significantly more prevalent in patients with NSAID-associated gastric ulcers than in patients with non-NSAID-associated gastric ulcer. The H. pylori infection rate was significantly lower in NSAID associated gastric ulcer patients than in non-NSAID-associated gastric ulcer patients (48% vs 96%, P<0.001). In the NSAID-associated gastric ulcer group, the prevalence of H. pylori infection was significantly lower in patients with ulcers in the antrum than in those with ulcers in the angulus or corpus (25% vs 77.3%, P<0.001). CONCLUSIONS: In contrast to non-NSAID-associated gastric ulcers, NSAID associated gastric ulcers frequently occur in the antrum with bleeding. The rate of H. pylori infection in NSAID-associated gastric ulcers is significantly lower than that in non-NSAID-associated gastric ulcers. PMID- 16706820 TI - A pilot study to evaluate a new combination therapy for gastric ulcer: Helicobacter pylori eradication therapy followed by gastroprotective treatment with rebamipide. AB - BACKGROUND AND AIM: Controversies remain over the need for antiulcer treatment following 1-week eradication triple therapy for Helicobacter pylori-positive peptic ulcers. The usefulness of combination therapy for gastric ulcers in Japanese patients, which consists of H. pylori eradication followed by gastroprotective therapy with rebamipide, was therefore evaluated. METHODS: The study was conducted in 52 H. pylori-positive patients with an endoscopically proven open gastric ulcer. All patients received 1-week triple therapy (lansoprazole, amoxicillin and clarithromycin) followed by 7-week rebamipide therapy. After completion of the combination therapy, all patients underwent evaluation of ulcer healing by endoscopy, gastric ulcer symptoms and H. pylori eradication by rapid urease test and (13)C-urea breath test. RESULTS: The ulcer healing rates were 85.7% (36/42) at 8 weeks, 83.3% (30/36) in eradicated patients and 100% (6/6) in non-eradicated patients. The overall gastrointestinal symptom free rate improved from 19.0% at baseline to 88.1% at 8 weeks. H. pylori was effectively eradicated in 85.7% (36/42) of patients. CONCLUSIONS: The results suggested that the combination therapy for open gastric ulcer was safe, well tolerated and effective. However, data from a double-blind placebo-controlled study is necessary to confirm these findings. PMID- 16706822 TI - Predictors of fibrosis in Asian patients with non-alcoholic steatohepatitis. AB - BACKGROUND AND AIM: Non-alcoholic steatohepatitis (NASH) is increasingly recognized as an important cause of chronic liver disease. However, data on Asians with NASH is lacking in the literature. The aim of the present study was to describe the clinical, biochemical and histological characteristics of NASH in Asians and to determine the predictors for septal fibrosis. METHOD: Sixty consecutive patients aged over 18 years with elevated serum alanine transferase, sonographic evidence of steatosis, and consent for liver biopsy were included. Patients with chronic hepatitis B or C, alcoholic, autoimmune, genetic, or drug induced liver disease were excluded. Clinical, biochemical and histological variables were tested for association with septal liver fibrosis (F2/3). RESULTS: Median age of the cohort was 45.5 years (range 21-75 years) and 63% were male. Ninety percent of patients were obese (body mass index [BMI]>or= 25), 70% had hypertriglyceridemia, 68% had hypercholesterolemia, 58% had metabolic syndrome, 53% had hypertension, 47% had diabetes mellitus (DM), and 18% had obstructive sleep apnea. Sixty-eight percent had gamma-glutamyl transferase (GGT) >or= 2 x upper limit of normal (ULN), 55% had alanine aminotransferase (ALT) >or= 2 x ULN, and 23% had aspartate aminotransferase (AST) >or= 2 x ULN. Of the 40 non-diabetic patients undergoing oral glucose tolerance testing, 45% had normal tests, 30% had impaired glucose tolerance, 23% DM, and 2% impaired fasting glucose. Eighteen patients (30%) had septal fibrosis (F2/3), but none had cirrhosis. Necroinflammatory grade >or= 2 (odds ratio [OR] 13), AST >or= 2 x ULN (OR 5.3) and DM (OR 5) were significantly and independently correlated with septal fibrosis. CONCLUSION: Septal fibrosis is common in Asians with NASH. Necroinflammatory grade >or= 2, AST >or= 2 x ULN and DM are independent predictors for septal fibrosis. PMID- 16706821 TI - Role of Helicobacter pylori virulence factor and genotypes in non-ulcer dyspepsia. AB - BACKGROUND AND AIM: The role of Helicobacter pylori (HP) in non-ulcer dyspepsia is debatable. Eradicating HP will help a small group of non-ulcer dyspeptic patients. However, it is unclear which subgroup of patients will benefit from eradication therapy. The aim of the present study was to compare the cagA and cagE status, as well as vacA genotypes, of HP in non-ulcer dyspeptic patients who responded successfully to eradication therapy compared with those patients who did not. METHODS: Consecutive patients with moderate to severe (Likert 2 or 3) non-ulcer dyspepsia with HP were recruited prospectively. Gastric biopsies were taken, DNA extracted and polymerase chain reaction performed to determine the cagA and cagE status and vacA alleles. Eradication therapy was offered until HP was eradicated successfully. The HP status was checked 1 month after eradication therapy using the [(13)C]-urea breath test. All patients were assessed by one interviewer using Gastrointestinal Symptom Rating Scale (GSRS), a four-point Likert scale, and SF-36 for quality of life over 12 months. Treatment success was defined as minimal or no symptoms (Likert 1 or 0). The cagA, cagE and vacA status was blinded to the investigators until completion of the study. RESULTS: Seventy three patients (36 males, 37 females) were recruited to the study. The mean+/-SD patient age was 53.38+/-12.09 years. When the 36 patients who improved (group A) were compared with the 37 (group B) who did not, no significant difference was found in the cagE positive rate (55.6 vs 43.2%, respectively; P=0.638), cagA positive rate (83.1 vs 73.0%, respectively; P=0.247), vacA m1 versus m2 subtype (84.0 vs 55.6%, respectively; P=0.472) or vacA s1a versus s1c (39.4 vs 57.1%, respectively; P=0.166). There was also no significant difference noted in the SF 36 scores between the two groups after the conclusion of eradication therapy. CONCLUSIONS: Stratification of HP genotypes and virulence factor has no significant impact on the treatment success of non-ulcer dyspepsia. PMID- 16706823 TI - Clinical features of hepatocellular carcinoma that occur after sustained virological response to interferon for chronic hepatitis C. AB - BACKGROUND AND AIM: This study investigated the clinical features of hepatocellular carcinoma in patients with sustained virological response to interferon for hepatitis C viral (HCV) infection. METHODS: A total of 7,715 patients with HCV infection were treated with interferon and followed up for more than 1 year after withdrawal of interferon in 64 Japanese hospitals and clinics between July 1988 and August 2001. Sustained virological response was obtained in 2,515 (32.6%) patients. Of these 2,515 patients, clinical data were collected for 38 patients in whom hepatocellular carcinoma developed. Sustained virological response was defined as HCV RNA negativity more than 6 months after the termination of interferon. RESULTS: All patients were HCV RNA negative at the time of diagnosis of hepatocellular carcinoma. The median period until the detection of hepatocellular carcinoma was 4.7 years (range 1.4-9.0 years). There were significant improvements in hepatic function including serum albumin, aspartate aminotransferase, alanine aminotransferase, indocyanine green test, platelet count and histological activity grade in comparison with those before interferon therapy and at the onset of hepatocellular carcinoma. The maximum tumor size in patients without medical follow-up for 1 year or more (median: 60 mm) was significantly larger than in patients who were periodically followed up for 6 months or less (median: 25 mm) (P=0.002). CONCLUSIONS: The present findings emphasize the importance of regular medical follow up of patients with HCV infection, as even patients showing a sustained virological response to interferon and in whom hepatic function has improved have the potential to develop hepatocellular carcinoma. PMID- 16706824 TI - Antiproliferative effects of 5-fluorouracil and interferon-alpha in combination on a hepatocellular carcinoma cell line in vitro and in vivo. AB - BACKGROUND AND AIM: We investigated the antiproliferative effects of interferon alpha (IFN-alpha) and 5-fluorouracil (5-FU) in combination on a hepatocellular carcinoma (HCC) cell line. METHOD: In the in vitro study, IFN-alpha and/or 5-FU was added to the culture of the poorly differentiated-type HCC cell line, HAK-1B, and their antiproliferative effects and additional or synergic effects in combination treatment were examined. In the in vivo study, HAK-1B cells were transplanted into nude mice and the changes in tumor volume and weight, apoptosis, BrdU and cyclin A positive cells, and artery-like blood vessels were investigated. Expressions of angiogenesis factors and IFN-alpha receptor (IFNAR 2) were examined in the developed tumors. RESULTS: In vitro growth of HAK-1B cells was suppressed dose-dependently to 5-FU, but the addition of IFN-alpha did not induce additional or synergic effects. In vivo growth in terms of tumor diameter and weight was suppressed at most in the IFN-alpha + 5-FU (combination) group, that is, the tumor volume became 29.3% and the tumor weight became 54.7% of the control. In the combination group, numbers of BrdU-positive S-phase cells and cyclin A positive cells increased together with the increase in apoptotic cells, but there was no significant relation between the tumor shrinkage effects and angiogenesis factors or artery-like blood vessels. In the combination group, INFAR-2 decreased significantly in comparison to the other groups. CONCLUSION: The synergic growth-suppression effects in the current in vivo study using the combination treatment are attributable to the enhanced induction of S-phase arrest and of apoptosis. PMID- 16706825 TI - Prevalence and risk factors of non-alcoholic fatty liver disease among Korean adults. AB - BACKGROUND AND AIM: Differences in the prevalence of non-alcoholic fatty liver disease (NAFLD) between Eastern and Western populations are primarily attributable to differences in definitions and biased population selection. Thus, the aim of the present study was to accurately determine the actual prevalence of NAFLD by sonography and to characterize the risk factors for NAFLD. METHODS: The present cross-sectional study was performed with data obtained from 6,648 subjects, all of whom were older than 20 years of age (3,530 men and 3,118 women). The term 'non-drinker' was applied to men who consumed less than 30 g alcohol/day and to women who consumed less than 20 g alcohol/day. Non-alcoholic fatty liver disease was defined as a sonographically detected fatty liver in the absence of viral hepatitis in a non-drinker. RESULTS: Of the 1,613 subjects who were diagnosed with sonographic fatty liver, 1,240 were non-drinkers and had no viral hepatitis. Overall, the unadjusted and age-adjusted prevalences of NAFLD were 18.7% (23% in men, 13.7% in women) and 16.1% (21.6% in men, 11.2% in women), respectively. Multivariate analysis revealed that several risk factors were profoundly associated with the prevalence of NAFLD, including obesity, insulin resistance, hyperlipidemia and hyperglycemia in both genders, as well as age, menopausal status and estrogen medication in women only. CONCLUSIONS: These results demonstrate that the prevalence of NAFLD in Korean adults, according to sonographic surveys, is comparable to that seen in more developed countries. From the perspective of increasing obesity, the high prevalence rates noted in the study may herald an increased burden of chronic liver disease in the Korean population. PMID- 16706826 TI - Role of transferrin receptor 2 in hepatic accumulation of iron in patients with chronic hepatitis C. AB - BACKGROUND AND AIM: Iron deposition in the liver is a common finding in patients with chronic hepatitis C (CH-C). The mechanism of this hepatic accumulation of iron is not completely understood. This study assessed if the protein expression of transferrin receptor 2 (TfR2) is upregulated in the liver of patients with CH C and if TfR2 protein mediates iron accumulation during hepatitis C virus (HCV) infection. METHOD: Liver specimens from patients with CH-C that underwent interferon (IFN) therapy (n=23) and from patients with CH-B (n=18) were evaluated. Hepatic expression of TfR2 protein was analyzed by immunohistochemistry. Total hepatic iron score (THIS) was evaluated by Prussian blue staining. RESULTS: TfR2 protein was expressed in the cell membrane and cytosol of hepatocytes. Cytosol TfR2 protein was found to co-localize with Tf. THIS (P=0.0198) and hepatic TfR2 (P=0.0047) expression were significantly higher in CH-C than in CH-B. The change in THIS values (rho=0.580, P=0.0079) and the grade of histological activity (rho=0.444, P=0.0373) were significantly correlated with changes in TfR2 expression after IFN therapy. CONCLUSIONS: The protein expression of TfR2 is significantly associated with iron deposition in the liver in patients with CH-C. HCV infection may affect the hepatic expression of TfR2, leading to iron accumulation in the liver. PMID- 16706827 TI - Expression of vascular endothelial growth factor-C in human hepatocellular carcinoma. AB - BACKGROUND/AIMS: Vascular endothelial growth factor-C (VEGF-C) is thought to be an important factor in tumor angiogenesis/lymphangiogenesis, but its role in hepatocellular carcinoma (HCC) has not yet been fully investigated. METHODS: We immunohistochemically examined VEGF-C expression in surgically resected tissues of 90 HCC. RESULTS: In the 78 HCC with a single histological grade, VEGF-C expression was significantly stronger in poorly differentiated HCC than in well- (P=0.003) or moderately differentiated HCC (P=0.0002). A 'nodule-in-nodule' case presented VEGF-A expression in the well-differentiated component and VEGF-C expression in the moderately-poorly differentiated component. According to nodular diameter, VEGF-C expression was significantly higher in nodules of 3.0 cm or larger (P=0.0263). Extrahepatic metastases seen in seven cases expressed VEGF C. In 20 of the 28 cases who were able to be followed up, the frequency of intrahepatic recurrence tended to be higher and extrahepatic metastasis was significantly higher in the cases who had VEGF-C expression in the tumor casts of the intrahepatic portal/hepatic vein branches than other cases without the expression (P=0.0139). Disease-free survival time tended to be shorter in cases with VEGF-C expression in tumor casts of the portal/hepatic vein than in those without VEGF-C expression (P=0.053; log-rank test). CONCLUSIONS: VEGF-C expression is related to the progression of HCC, and VEGF-C expression in tumor casts of the intrahepatic portal/hepatic vein is considered to be a factor indicating recurrence/metastasis sites. PMID- 16706828 TI - Chemokine receptor CCR6 as a prognostic factor after hepatic resection for hepatocellular carcinoma. AB - BACKGROUND AND AIMS: Chemokines and their receptors have recently been shown to have major roles in cancer metastasis. The aim of this study was to determine whether the interaction between chemokine receptor 6 (CCR6) and its ligand, macrophage inflammatory protein-3 alpha (MIP-3alpha), correlates with metastasis of hepatocellular carcinoma (HCC). METHODS: To observe the reaction of CCR6 expressed cancer cells to MIP-3alpha stimulation, chemotactic and actin polymerization assays for both CCR6 high cells (HepG2) and CCR6 low cells (MCF-7) were performed. CCR6 mRNA levels in tumor specimens from 30 HCC patients were quantified by real-time polymerase chain reaction. Patients were classified into two groups, high (>or= 20 copies; n=10) CCR6 and low (<20 copies; n=20) CCR6 on the basis of CCR6 expression, and the groups were compared with respect to clinicopathological features. RESULTS: When HepG2 cells (CCR6 high) were stimulated with MIP-3alpha, they migrated in a dose-dependent manner, and formation of pseudopodia was observed. These phenomena were not observed in the CCR6 low cells. The incidence of intrahepatic metastasis was higher in the high CCR6 expression group than in the low CCR6 expression group (P<0.05). Disease free survival was significantly poorer in the high CCR6 expression group than in the low CCR6 expression group (P<0.05). CONCLUSIONS: It was indicated that CCR6 might be associated with intrahepatic metastasis of HCC and might be able to become one of the prognostic factor after hepatic resection for HCC. PMID- 16706829 TI - Long-term prognosis of nonalcoholic fatty liver disease: is pharmacological therapy actually necessary? AB - BACKGROUND AND AIM: Nonalcoholic fatty liver disease (NAFLD) comprises a wide spectrum of liver injury, ranging from steatosis and steatohepatitis to cirrhosis. Reasons for the different natural course in individuals with NAFLD are still unclear. The aim of this study was to describe the natural course of disease in individuals with NAFLD who did not receive pharmacological therapy. METHODS: A total of 27 individuals with NAFLD (male/female ratio: 10/17, mean age 49.7 years) were prospectively enrolled. Management after diagnosis consisted of establishment of an appropriate diet and exercise (walking and jogging) program, treatment of associated metabolic conditions such as diabetes and dyslipidemia, and discontinuation of potentially hepatotoxic drugs if the patient was taking these. Liver tests were performed at diagnosis and at 3-month intervals during the follow-up period. Mean follow-up period was 43.3 months (range 36-49 months). RESULTS: From baseline to the end of the follow-up period, although there was no significant difference observed in terms of the mean body mass index, serum aminotransferase levels significantly improved (48.8+/-29.9 U/L to 31.6+/-16.0 U/L for aspartate aminotransferase [AST] and 66.3+/-38.3 U/L to 39.6+/-22.9 U/L for alanine aminotransferase [ALT]; P<0.05). No significant differences in platelet counts, serum albumin level or prothrombin time were observed (P>0.05). No patient developed signs of advanced liver disease during the follow-up period. CONCLUSION: A treatment strategy comprising diet, exercise and management of associated metabolic conditions is associated with improvement in aminotransferases among patients with NAFLD. Further investigation is needed to examine the long-term efficacy of this approach on liver histology and clinical outcomes. PMID- 16706830 TI - Rodent nutritional model of steatohepatitis: effects of endotoxin (lipopolysaccharide) and tumor necrosis factor alpha deficiency. AB - BACKGROUND AND AIMS: Intestinal endotoxin (lipopolysaccharide) is thought to contribute to liver injury in both alcoholic and nonalcoholic steatohepatitis (NASH). Tumor necrosis factor alpha (TNFalpha) is an important mediator of this process and is considered central to the inflammatory response in NASH. This study aimed to investigate the effects of lipopolysaccharide on liver injury in the methionine choline deficient (MCD) nutritional model of NASH, and to determine if TNFalpha is required for the development of steatohepatitis in this model. METHOD: Male C57/BL6 mice received a MCD diet for 4 weeks, whilst a control group received an identical diet supplemented with 0.2% choline bitartrate and 0.3% methionine. At 4 weeks, mice received either an intraperitoneal injection of lipopolysaccharide (0.5 microg/g body mass) or sterile saline, and were killed 24 h thereafter. In a separate study, TNFalpha knockout and wild type C57BL/6 mice received either MCD or control diets for 4 weeks. Serum transaminase levels, liver histology (steatosis, inflammation and apoptosis), hepatic triglyceride concentration and hepatic lipid peroxidation products (conjugated dienes, lipid hydroperoxides and thiobarbituric reactive substances, free and total) were evaluated. RESULTS: Intraperitoneal administration of lipopolysaccharide augmented serum alanine aminotransferase (ALT) levels (P<0.02), hepatic inflammation (P<0.025), apoptosis (P<0.01) and free thiobarbituric acid reactive substances (P<0.025) in MCD mice. TNFalpha knockout mice fed the MCD diet developed steatohepatitis with histological and biochemical changes similar to those seen in wild type counterparts. CONCLUSIONS: Lipopolysaccharide augments liver injury in MCD mice, and TNFalpha is not required for the development of steatohepatitis in MCD mice. PMID- 16706831 TI - Origin of neovascular structure in an early stage of hepatocellular carcinoma: study of alpha-smooth muscle actin immunohistochemistry in serial thin sections of surgically resected cancer. AB - BACKGROUND: To elucidate the origin of the neovascular structure found in well differentiated hepatocellular carcinoma (HCC), an immunohistochemical study was performed on sequential thin section specimens. METHOD: Eleven surgically resected specimens of well-differentiated HCC were analyzed for neovascular structure using monoclonal alpha-smooth muscle actin (alpha-SMA) antibody. Each paraffin specimen was serially sliced to a thickness of 3 microm for immunohistochemistry. When a ring-shaped structure was found unrelated to portal triads on alpha-SMA staining, it was regarded as abnormal neovascularity (non triadal vessel or unaccompanied vessel). RESULTS: All of the 11 liver cancers had thin-walled, round- or oval-shaped non-triadal vessels in their well differentiated parts. Immunohistochemistry of serial thin sections of HCC showed that these non-triadal vessels were connected to portal veins in portal triads in well-differentiated cancer in a total of nine patients (81.8%). This type of neovascular structure found in a well-differentiated cancer seemed to be a surviving portal vein among diminishing and disappearing arteries and bile ducts. All 11 tumors showed isovascular staining on ordinary digital subtraction angiography, and four of the tumors showed negative enhancement on intra-arterial carbon dioxide-enhanced ultrasonography or computerized tomographic (CT) hepatic arteriography, suggesting a relative arterial blood scarcity in the tumor nodules. CONCLUSION: At an early stage of HCC, non-triadal vessels originate from ordinary portal veins in intratumoral portal triads. This fact sufficiently explains the reason why a well-differentiated liver cancer can sometimes show arterial blood paucity on CT arteriography or enhanced ultrasonography. PMID- 16706832 TI - Effect of exercise and dietary modification on serum aminotransferase levels in patients with nonalcoholic steatohepatitis. AB - BACKGROUND: Nonalcoholic steatohepatitis (NASH) is commonly associated with overweight and insulin resistance. Aerobic exercise is known to reduce insulin resistance. We studied the effect of regular aerobic exercise on serum aminotransferase levels in patients with NASH. METHODS: Sixty-five (mean age 38.7+/-9.5 years; 46 [78%] males) out of 94 patients diagnosed with NASH participated in the study. Each patient was advised regular aerobic exercise for 30 min duration per day, and trained to achieve a heart rate of 60-70% of his/her maximal heart rate for at least 5 days a week. In addition, those with a high body mass index (BMI) were advised a moderately energy-restricted diet. Patients were followed up monthly for at least 3 months by BMI, waist-hip ratio (WHR), waist circumference (WC), aspartate aminotransferase (AST) and alanine aminotransferase (ALT). RESULTS: Forty-three (72.8%) patients had a high BMI. Central obesity (WHR >or=0.90 cm in men and >or=0.85 cm in women) was present in 58 (98.3%) patients and metabolic syndrome in 12 (20.3%) patients. In the 44 patients who complied regularly with the exercise program, serum ALT normalized in 20 (45%; P<0.05), and mean AST and ALT values declined from 70.5 and 104.0 41.5 (P<0.001) and 63.2 (P<0.001), respectively. Overweight patients lost a mean 3.1 kg (range 0-13 kg) weight and showed a decline in BMI (28.7 vs 27.5; P<0.001) and WC (99.4 cm vs 96.1 cm; P=0.001). Serum ALT did not normalize in any of the 15 patients who failed to comply with the exercise program; their pre- and post AST (82.8 and 81.2, respectively) and ALT (98.0 and 96.1, respectively) levels, BMI (27.5 and 27.6, respectively) and WHR (0.99 cm and 0.99 cm, respectively) did not show any significant change at 3 months, and also for an extended mean follow up period of 5.3 months. CONCLUSION: Moderate intensity aerobic exercise helps in normalizing ALT levels in patients with NASH. PMID- 16706833 TI - Semaphorin3A regulates synaptic function of differentiated hippocampal neurons. AB - Semaphorins are major chemorepellents for developing neuronal projections. Their persistent expression at adult stages suggests that they may contribute to the functioning of neuronal circuits. We investigated the functional properties of semaphorin3A (Sema3A) in adult hippocampal neurons, and report that exogenous application of this cue decreases the efficacy of synaptic transmission evoked in the CA1 region of hippocampal slices. In situ hybridization, imaging and biochemical techniques showed that the Sema3A receptor component neuropilin-1 is present at hippocampal synapses and localizes in the presynaptic membrane. In differentiated cultured hippocampal neurons, Sema3A elicited Erk1/2 phosphorylation in somata and neuritic compartments. Furthermore, Sema3A application resulted in a striking reduction of synaptophysin and postsynaptic density 95 puncta without affecting the axon diameter. These observations reveal novel functional potentialities for secreted semaphorins, which suggest that these cues could modulate the morphology and function of synapses in the adult brain. PMID- 16706834 TI - In vivo synaptic plasticity in the dentate gyrus of mice deficient in the neural cell adhesion molecule NCAM or its polysialic acid. AB - The neural cell adhesion molecule NCAM and its associated polysialic acid (PSA) play important roles in synaptic plasticity in the CA1 and/or CA3 regions of the hippocampus in vitro. Here, we address the question of whether NCAM and PSA are involved in regulation of synaptic transmission and plasticity also in vivo at synapses formed by entorhinal cortex axons in the dentate gyrus of mice anaesthetized with urethane. We show that basal synaptic transmission, measured as the slope of field excitatory postsynaptic potentials, was reduced strongly in mice lacking ST8SiaII/STX, the enzyme involved in polysialylation of NCAM in stem cell-derived immature granule cells, but not in mice deficient either in the NCAM glycoprotein or the enzyme ST8SiaIV/PST involved in polysialylation of NCAM in mature neurons. Strikingly, only mice deficient in NCAM, but not in PST or STX, were impaired in long-term potentiation (LTP) induced by theta-burst stimulation, suggesting that LTP in the dentate gyrus depends on the NCAM glycoprotein alone rather than on its associated PSA. As also patterns of synaptic activity during and immediately after induction of LTP were impaired in NCAM-deficient mice, it is likely that induction of LTP requires NCAM. These data are the first to describe that NCAM is necessary for induction of synaptic plasticity in identified synapses in vivo and suggest that polysialylation of NCAM expressed by immature granule cells in the dentate gyrus supports development of basal excitatory synaptic transmission in this region. PMID- 16706835 TI - Expression of trophinin and bystin identifies distinct cell types in the germinal zones of adult rat brain. AB - In the adult brain, the subventricular zone (SVZ) is one of the regions where active neurogenesis occurs. Relatively few specific markers are available to distinguish different types of cells in the SVZ and rostral migratory stream (RMS) of adult brain. Here, we showed that trophinin and bystin, both of which are required for early embryo implantation during development, were expressed in the SVZ and RMS of the adult rat brain, but not in the brain of embryos and early postnatal animals. Trophinin-expressing cells were immunopositive for both Ki-67 and nestin in the SVZ. Some of the trophinin-positive cells did not express either the type A cell marker polysialylated weakly adhesive form of the neural cell adhesion molecule (PSA-NCAM) or the type B cell marker glial fibrillary acidic protein (GFAP). Double-label immunohistochemistry revealed that bystin positive cells co-expressed GFAP, Ki-67 and nestin, but not PSA-NCAM, suggesting that they are likely type B cells. Intracerebroventricular infusion of cytosine beta-d-arabiofuranoside (Ara-C) eliminated trophinin-positive cells in the SVZ. Following its depletion, however, the remaining bystin-positive cells continued to divide and generate actively dividing trophinin-positive cells that were negative for PSA-NCAM, leading to reconstruction of SVZ network. These characteristics indicate that this subset of trophinin-positive cells in the SVZ is type C cells. Conversely in the RMS, trophinin co-localized with nestin and PSA-NCAM, suggesting that it is expressed in neuroblasts. Cultured neural precursor cells derived from the adult SVZ also expressed both trophinin and bystin. These findings provide insight into the molecular basis of adult neurogenesis in the SVZ and RMS. PMID- 16706836 TI - Requirement for Id1 in opioid-induced oligodendrogenesis in cultured adult rat hippocampal progenitors. AB - Growth factors and peptides playing important roles during early development of the central nervous system have also been shown to maintain their regulation of cell genesis in the adult brain. We have previously described that endogenous opioids, expressed in the developing hippocampus, regulate proliferation and differentiation in the adult rat hippocampus. The aim of this study was to investigate the effects of the opioid beta-endorphin on gene expression and glial differentiation in cultures of adult rat hippocampal progenitors (AHPs). Changes in gene expression after stimulation of AHPs with beta-endorphin for 48 h were investigated using cDNA arrays. Confirmation experiments verified that stimulation with beta-endorphin increased the mRNA levels of myelin basic protein, glutathione S-transferase pi, c-junD and rab16 (P < 0.05), genes that are associated with oligodendrogenesis. Furthermore, beta-endorphin increased the levels of Id1, but not Id3, mRNA on the arrays. Incubation of AHPs with beta endorphin resulted in a threefold increase in oligodendrogenesis (P < 0.01) but no significant change in astrogliogenesis. No effect on oligodendrogenesis was observed in the presence of the opioid antagonist naloxone. Coincubation of beta endorphin with Id1 antisense oligonucleotides for 10 days also entirely blocked the induced oligodendrogenesis in our AHP cultures. Moreover, a subpopulation of AHPs (25%) showed nuclear expression of the proneural transcriptional activator Mash1 that was reduced to approximately 5% of the cells when exposed to beta endorphin. We suggest a requirement for Id1 in opioid-induced oligodendrogenesis in cultured AHPs possibly acting on opioid-responsive AHPs expressing the proneural transcriptional activator Mash1. PMID- 16706837 TI - Notch controls proliferation and differentiation of stem cells in a dose dependent manner. AB - Self-renewal and differentiation of CNS stem cells are regulated by still poorly understood cell-cell interactions. Notch is a well-known cell surface protein that can promote both cell cycle progression and mitotic arrest but the molecular mechanism controlling these opposite effects is unknown. Here we demonstrate that, in CNS stem cells, the level of active Notch1 determines the cellular response. Specifically, low levels of the active form of Notch1 promote proliferation whereas high levels lead to growth arrest. Here we provide the first evidence that Notch effects on proliferation and differentiation are a function of dose, and propose a hypothesis on how oncogenes may also act as tumor suppressors. PMID- 16706838 TI - Ca1.2 and CaV1.3 neuronal L-type calcium channels: differential targeting and signaling to pCREB. AB - Neurons express multiple types of voltage-gated calcium (Ca2+) channels. Two subtypes of neuronal L-type Ca2+ channels are encoded by CaV1.2 and CaV1.3 pore forming subunits. To compare targeting of CaV1.2 and CaV1.3 L-type Ca2+ channels, we transfected rat hippocampal neuronal cultures with surface-epitope-tagged sHA CaV1.2 or sHA-CaV1.3a constructs and found that: (i) both sHA-CaV1.2 and sHA CaV1.3a form clusters on the neuronal plasma membrane surface; (ii) when compared with sHA-CaV1.2 surface clusters, the sHA-CaV1.3a surface clusters were 10% larger and 25% brighter, but 35% less abundant; (iii) 81% of sHA-CaV1.2 surface clusters, but only 48% of sHA-CaV1.3a surface clusters, co-localized with synapsin clusters; (iv) co-expression with GFP-Shank-1B had no significant effect on sHA-CaV1.2 surface clusters, but promoted formation and synaptic localization of sHA-CaV1.3a surface clusters. In experiments with dihydropyridine-resistant CaV1.2 and CaV1.3a mutants we demonstrated that CaV1.3a L-type Ca2+ channels preferentially mediate nuclear pCREB signaling in hippocampal neurons at low, but not at high, levels of stimulation. In experiments with primary neuronal cultures from CaV1.3 knockout mice we discovered that CaV1.3 channels play a more important role in pCREB signaling in striatal medium spiny neurons than in hippocampal neurons. Our results provide novel insights into the function of CaV1.2 and CaV1.3 L-type Ca2+ channels in the brain. PMID- 16706839 TI - Two PEST-like motifs regulate Ca2+/calpain-mediated cleavage of the CaVbeta3 subunit and provide important determinants for neuronal Ca2+ channel activity. AB - An increase in intracellular Ca2+ due to voltage-gated Ca2+ (CaV) channel opening represents an important trigger for a number of second-messenger-mediated effects ranging from neurotransmitter release to gene activation. Ca2+ entry occurs through the principal pore-forming protein but several ancillary subunits are known to more precisely tune ion influx. Among them, the CaVbeta subunits are perhaps the most important, given that they largely influence the biophysical and pharmacological properties of the channel. Notably, several functional features may be associated with specific structural regions of the CaVbeta subunits emphasizing the relevance of intramolecular domains in the physiology of these proteins. In the current report, we show that CaVbeta3 contains two PEST motifs and undergoes Ca2+ -dependent degradation which can be prevented by the specific calpain inhibitor calpeptin. Using mutant constructs lacking the PEST motifs, we present evidence that they are necessary for the cleavage of CaVbeta3 by calpain. Furthermore, the deletion of the PEST sequences did not affect the binding of CaVbeta3 to the ion-conducting CaV2.2 subunit and, when expressed in human embryonic kidney-293 cells, the PEST motif-deleted CaVbeta3 significantly increased whole-cell current density and retarded channel inactivation. Consistent with this observation, calpeptin treatment of human embryonic kidney 293 cells expressing wild-type CaVbeta3 resulted in an increase in current amplitude. Together, these findings suggest that calpain-mediated CaVbeta3 proteolysis may be an essential process for Ca2+ channel functional regulation. PMID- 16706840 TI - T-type CaV3.3 calcium channels produce spontaneous low-threshold action potentials and intracellular calcium oscillations. AB - The precise contribution of T-type Ca2+ channels in generating action potentials (APs), burst firing and intracellular Ca2+ signals needs further elucidation. Here, we show that CaV3.3 channels can trigger repetitive APs, generating spontaneous membrane potential oscillations (MPOs), and a concomitant increase in the intracellular Ca2+ concentration ([Ca2+]i) when overexpressed in NG108-15 cells. MPOs were dependent on CaV3.3 channel activity given that they were recorded from a potential range of -55 to -70 mV, blocked by nickel and mibefradil, as well as by low external Ca2+ concentration. APs of distinct duration were recorded: short APs (sAP) or prolonged APs (pAP) with a plateau potential near -40 mV. The voltage-dependent properties of the CaV3.3 channels constrained the AP duration and the plateau potential was supported by sustained calcium current through CaV3.3 channels. The sustained current amplitude decreased when the resting holding potential was depolarized, thereby inducing a switch of AP shape from pAP to sAP. Duration of the [Ca2+]i oscillations was also closely related to the shape of APs. The CaV3.3 window current was the oscillation trigger as shown by shifting the CaV3.3 window current potential range as a result of increasing the external Ca2+ concentration, which resulted in a corresponding shift of the AP threshold. Overall, the data demonstrate that the CaV3.3 window current is critical in triggering intrinsic electrical and [Ca2+]i oscillations. The functional expression of CaV3.3 channels can generate spontaneous low-threshold calcium APs through its window current, indicating that CaV3.3 channels can play a primary role in pacemaker activity. PMID- 16706841 TI - Intrinsic bursting of immature CA3 pyramidal neurons and consequent giant depolarizing potentials are driven by a persistent Na+ current and terminated by a slow Ca2+ -activated K+ current. AB - The CA3 area of the mature hippocampus is known for its ability to generate intermittent network activity both in physiological and in pathological conditions. We have recently shown that in the early postnatal period, the intrinsic bursting of interconnected CA3 pyramidal neurons generates network events, which were originally called giant depolarizing potentials (GDPs). The voltage-dependent burst activity of individual pyramidal neurons is promoted by the well-known depolarizing action of endogenous GABA on immature neurons. In the present work, we show that a persistent Na+ current, I-Nap, accounts for the slow regenerative depolarization that triggers the intrinsic bursts in the neonatal rat CA3 pyramidal neurons (postnatal day 3-6), while a slow Ca2+ -activated K+ current, sI-K(Ca), is primarily responsible for the postburst slow afterhyperpolarization and consequent burst termination. In addition, we exploited pharmacological data obtained from intracellular recordings to study the mechanisms involved in network events recorded with field potential recordings. The data as a whole indicate that I-Nap and sI-K(Ca) are involved in the initiation and termination, respectively, of the pyramidal bursts and consequent network events underlying GDPs. PMID- 16706842 TI - A novel RNA binding protein that interacts with NMDA R1 mRNA: regulation by ethanol. AB - Excitatory NMDA receptors are an important target of ethanol. Chronic ethanol exposure, in vivo and in vitro, increases polypeptide levels of NR1 subunit, the key subunit of functional NMDA receptors. In vitro, chronic ethanol treatment increases the half-life of NR1 mRNA and this observation is dependent on new protein synthesis. The present study was undertaken to locate cis-acting region(s) within the NR1 3'-untranslated region (UTR) and identify NR1 3'-UTR binding trans-acting proteins expressed in mouse fetal cortical neurons. Utilizing RNA gel shift assays we identified a 156-nt cis-acting region that binds to polysomal trans-acting proteins. This binding was highly specific as inclusion of cyclophilin RNA or tRNA did not interfere with cis-trans interactions. Importantly, the 3'-UTR binding activity was significantly up regulated in the presence of ethanol. UV cross-link analysis detected three NR1 3'-UTR binding proteins and their molecular mass calculated by Northwestern analysis was approximately 88, 60 and 47 kDa, respectively. Northwestern analysis showed a significant up-regulation of the 88-kDa protein after chronic ethanol treatment. The 88-kDa protein was purified and identified by tandem mass spectrometry as the beta subunit of alpha glucosidase II (GIIbeta). That GIIbeta is indeed a trans-acting protein and binds specifically to 3'-UTR of NR1 mRNA was confirmed by RNA gel mobility supershift assays and immuno RT-PCR. Western blotting data established a significant increase of GIIbeta polypeptide in chronic ethanol-exposed fetal cortical neurons. We hypothesize that the identified cis-acting region and the associated RNA-binding proteins are important regulators of NR1 subunit gene expression. PMID- 16706843 TI - Rapid phosphorylation of histone H2A.X following ionotropic glutamate receptor activation. AB - Excessive activation of ionotropic glutamate receptors increases oxidative stress, contributing to the neuronal death observed following neurological insults such as ischemia and seizures. Post-translational histone modifications may be key mediators in the detection and repair of damage resulting from oxidative stress, including DNA damage, and may thus affect neuronal survival in the aftermath of insults characterized by excessive glutamate release. In non neuronal cells, phosphorylation of histone variant H2A.X (termed gamma-H2AX) occurs rapidly following DNA double-strand breaks. We investigated gamma-H2AX formation in rat cortical neurons (days in vitro 14) following activation of N methyl-D-aspartate (NMDA) or alpha-amino-3-hydroxyl-5-methyl-4-isoxazolepropionic acid (AMPA)/kainate glutamate receptors using fluorescent immunohistochemical techniques. Moreover, we evaluated the co-localization of gamma-H2AX 'foci' with Mre11, a double-strand break repair protein, to provide further evidence for the activation of this DNA damage response pathway. Here we show that minimally cytotoxic stimulation of ionotropic glutamate receptors was sufficient to evoke gamma-H2AX in neurons, and that NMDA-induced gamma-H2AX foci formation was attenuated by pretreatment with the antioxidant, Vitamin E, and the intracellular calcium chelator, BAPTA-AM. Moreover, a subset of gamma-H2AX foci co-localized with Mre11, indicating that at least a portion of gamma-H2AX foci is damage dependent. The extent of gamma-H2AX induction following glutamate receptor activation corresponded to the increases we observed following conventional DNA damaging agents [i.e. non-lethal doses of gamma-radiation (1 Gy) and hydrogen peroxide (10 microm)]. These data suggest that insults not necessarily resulting in neuronal death induce the DNA damage-evoked chromatin modification, gamma H2AX, and implicate a role for histone alterations in determining neuronal vulnerability following neurological insults. PMID- 16706844 TI - Spike-timing-dependent plasticity at resting and conditioned lateral perforant path synapses on granule cells in the dentate gyrus: different roles of N-methyl D-aspartate and group I metabotropic glutamate receptors. AB - We examined the mechanisms underlying spike-timing-dependent plasticity induction at resting and conditioned lateral perforant pathway (LPP) synapses in the rat dentate gyrus. Two stimulating electrodes were placed in the outer third of the molecular layer and in the granule cell layer in hippocampal slices to evoke field excitatory postsynaptic potentials (fEPSPs) and antidromic field somatic spikes (afSSs), respectively. Long-term potentiation (LTP) of LPP synapses was induced by paired stimulation with fEPSP preceding afSS. Reversal of the temporal order of fEPSP and afSS stimulation resulted in long-term depression (LTD). Induction of LTP or LTD was blocked by D,L-2-amino-5-phosphonopentanoic acid (AP5), showing that both effects were N-methyl-D-aspartate receptor (NMDAR) dependent. Induction of LTP was also blocked by inhibitors of calcium-calmodulin kinase II, protein kinase C or mitogen-activated/extracellular-signal regulated kinase, suggesting that these are downstream effectors of NMDAR activation, whereas induction of LTD was blocked by inhibitors of protein kinase C and protein phosphatase 2B. At LPP synapses previously potentiated by high-frequency stimulation or depressed by low-frequency stimulation, paired fEPSP-afSS stimulation resulted in 'de-depression' at depressed LPP synapses but had no effect on potentiated synapses, whereas reversal of the temporal order of fEPSP afSS stimulation resulted in 'de-potentiation' at potentiated synapses but had no effect on depressed synapses. Induction of de-depression and de-potentiation was unaffected by ap5 but was blocked by 2-methyl-6-(phenylethynyl) pyridine hydrochloride, a group I metabotropic glutamate receptor blocker, showing that both were NMDAR-independent but group I metabotropic glutamate receptor dependent. In conclusion, our results show that spike-timing-dependent plasticity can occur at both resting and conditioned LPP synapses, its induction in the former case being NMDAR-dependent and, in the latter, group I metabotropic glutamate receptor-dependent. PMID- 16706845 TI - Oxidative modulation of the transient potassium current IA by intracellular arachidonic acid in rat CA1 pyramidal neurons. AB - Oxidative stress affects cellular membrane lipids and proteins. Using whole-cell patch-clamp recording we demonstrate differential oxidative inhibition of voltage gated transient (IA) and delayed rectifier [IK(V)] K+ currents by arachidonic acid (AA) and H2O2 in CA1 neurons in hippocampal slice. We show that intracellular application of 1 pm AA or its non-metabolizable analog eicosatetraynoic acid (100 pm) reduced IA by approximately 42% but did not affect IK(V). AA shifted the voltage dependence of steady-state inactivation of IA by 12 mV to more negative potentials whereas the rate of inactivation was unchanged. Surprisingly, intracellular glutathione (GSH, 20 mm) enhanced the effect of AA on maximal IA (-62%) and with AA slowed inactivation of IA. The combination of GSH and extracellular ascorbate (0.4 mm) prevented reduction of IA by AA. Intracellular Trolox (a vitamin E analog, 10 microm) reduced IA by 61%and IK(V) by 39%. Like AA, intracellular Trolox caused a 10-mV left shift of IA steady state inactivation but Trolox and AA did not cause a shift when coapplied. Extracellular Trolox (100 microm) had no effects on IA. H2O2 (80 microm) reduced both IA and IK(V) in a GSH- and ascorbate-sensitive manner and slowed the rate of inactivation of IA by a factor of 2. Coapplication of H2O2 with GSH and extracellular ascorbate caused approximately 22 mV negative shifts of both steady state inactivation and activation. We conclude that AA is extremely potent in affecting IA by oxidative modifications. Antioxidants can augment these effects, probably by catalysis of the underlying reactions between oxidants and IA channel proteins. PMID- 16706846 TI - Cannabinoids modulate spontaneous neuronal activity and evoked inhibition of locus coeruleus noradrenergic neurons. AB - The noradrenergic pathway arising from the locus coeruleus (LC) is involved in the regulation of attention, arousal, cognitive processes and sleep. These physiological activities are affected by Cannabis exposure - both in humans and laboratory animals. In addition, exogenous cannabinoids, as well as pharmacological and genetic manipulation of the endocannabinoid system, are known to influence emotional states (e.g. anxiety) for which a contributory role of the LC-noradrenergic system has long been postulated. However, whether cannabinoid administration would affect the LC neuronal activity in vivo is still unknown. To this end, single-unit extracellular recordings were performed from LC noradrenergic cells in anaesthetized rats. Intravenous injection of both the synthetic cannabinoid agonist, WIN55212-2, and the main psychoactive principle of Cannabis, Delta9-tetrahydrocannabinol, dose-dependently increased the firing rate of LC noradrenergic neurons, with WIN55212-2 being the most efficacious. Similar results were obtained by the administration of these drugs into a lateral ventricle. Cannabinoid-induced stimulation of LC noradrenergic neuronal activity was counteracted by SR141716A, a cannabinoid receptor antagonist/reverse agonist, which by itself slightly reduced LC discharge rate. Moreover, WIN55212-2 suppressed the inhibition of noradrenergic cells produced by stimulation of the major gamma-aminobutyric acid (GABA)ergic afferent to the LC, the nucleus prepositus hypoglossi. Altogether, these findings suggest the involvement of noradrenergic pathways in some consequences of Cannabis intake (e.g. cognitive and attention deficits, anxiety reactions), as well as a role for cannabinoid receptors in basic brain activities sustaining arousal and emotional states. PMID- 16706847 TI - L-DOPA-induced dyskinesia in adult rats with a unilateral 6-OHDA lesion of dopamine neurons is paralleled by increased c-fos gene expression in the subthalamic nucleus. AB - Levodopa (L-DOPA), the metabolic precursor of dopamine, is widely used as a pharmacological agent for the symptomatic treatment of Parkinson's disease. However, long-term L-DOPA use results in abnormal involuntary movements such as dyskinesias. There is evidence that abnormal cell signaling in the basal ganglia is involved in L-DOPA-induced dyskinesia. The subthalamic nucleus (STN) plays a key role in the circuitry of the basal ganglia and in the pathophysiology of Parkinson's disease. However, the contribution of the STN to L-DOPA-induced dyskinesias remains unclear. The objective of this work was to study the effects of acute or chronic systemic administration of L-DOPA to adult rats with a unilateral 6-hydroxydopamine (6-OHDA) lesion of dopamine neurons on c-fos expression in the STN and test the hypothesis that these effects correlate with L DOPA-induced dyskinesias. c-fos mRNA expression was measured in the STN by in situ hybridization histochemistry at the single cell level. Our results confirm earlier evidence that the chronic administration of L-DOPA to rats with a unilateral 6-OHDA lesion increases c-fos expression in the STN. We also report that c-fos expression can be increased following an acute injection of L-DOPA to 6-OHDA-lesioned rats but not following a chronic injection of L-DOPA to sham operated, unlesioned rats. Finally, we provide evidence that the occurrence and severity of dyskinesia is correlated with c-fos mRNA levels in the ipsilateral STN. These results suggest that altered cell signaling in the STN is involved in some of the behavioral effects induced by systemic L-DOPA administration. PMID- 16706848 TI - Compensatory changes in the hippocampus of somatostatin knockout mice: upregulation of somatostatin receptor 2 and its function in the control of bursting activity and synaptic transmission. AB - Somatostatin-14 (SRIF) co-localizes with gamma-aminobutyric acid (GABA) in the hippocampus and regulates neuronal excitability. A role of SRIF in the control of seizures has been proposed, although its exact contribution requires some clarification. In particular, SRIF knockout (KO) mice do not exhibit spontaneous seizures, indicating that compensatory changes may occur in KO. In the KO hippocampus, we examined whether specific SRIF receptors and/or the cognate peptide cortistatin-14 (CST) compensate for the absence of SRIF. We found increased levels of both sst2 receptors (sst2) and CST, and we explored the functional consequences of sst2 compensation on bursting activity and synaptic responses in hippocampal slices. Bursting was decreased by SRIF in wild-type (WT) mice, but it was not affected by either CST or sst2 agonist and antagonist. sst4 agonist increased bursting frequency in either WT or KO. In WT, but not in KO, its effects were blocked by agonizing or antagonizing sst2, suggesting that sst2 and sst4 are functionally coupled in the WT hippocampus. Bursting was reduced in KO as compared with WT and was increased upon application of sst2 antagonist, while SRIF, CST and sst2 agonist had no effect. At the synaptic level, we observed that in WT, SRIF decreased excitatory postsynaptic potentials which were, in contrast, increased by sst2 antagonist in KO. We conclude that sst2 compensates for SRIF absence and that its upregulation is responsible for reduced bursting and decreased excitatory transmission in KO mice. We suggest that a critical density of sst2 is needed to control hippocampal activity. PMID- 16706849 TI - Sex- and cell-type-specific patterns of GABAA receptor and estradiol-mediated signaling in the immature rat substantia nigra. AB - The substantia nigra pars reticulata (SNR) is involved in movement and seizure control. In male but not female postnatal day 15 (PN15) rats, GABAA receptor agonists depolarize the SNR neurons and increase the expression of the calcium regulated gene KCC2 (potassium/chloride cotransporter). Moreover, in PN15 rat SNR, 7beta-estradiol down-regulates KCC2 expression only in the presence of depolarizing GABAA receptor responses. The hypothesis tested here was that GABAA receptors and estradiol also regulate the expression of the phosphorylated form of the transcription factor cAMP responsive element binding protein (phosphoCREB), in PN15 rat SNR and substantia nigra pars compacta (SNC). Rats were injected with muscimol or 17beta-estradiol or their vehicles, and killed 1 h later. Sections were stained with an antibody specific for phosphoCREB alone or counterstained with either tyrosine hydroxylase (TH)- or parvalbumin (PRV) specific antibodies. Muscimol increased phosphoCREB-ir in male but not in female SN neurons. Using gramicidin perforated patch clamp of PN14-15 SNC neuron, it was shown that muscimol bath application depolarized male SNC neurons but did not significantly alter membrane potential in females. In males, 17beta-estradiol decreased phosphoCREB expression in all studied cell types. In females, 17beta estradiol did not influence phosphoCREB expression in PRV-ir SNR cells, but increased it in the dopaminergic SN neurons. These data suggest that GABAA receptor activation and estradiol promote the sexual differentiation of the SN in a cell-type-specific manner, by influencing calcium-regulated gene transcription, and therefore promoting the acquisition of sex-specific roles of the SN in movement and seizure control. PMID- 16706850 TI - Maturation of otolith-related brainstem neurons in the detection of vertical linear acceleration in rats. AB - To investigate the critical maturation time of otolith-related neurons in processing vertical orientations, rats (postnatal day 4 to adults) were studied for functional activation of c-fos expression in brainstem neurons by immuno /hybridization histochemistry. Conscious rats were subjected to sinusoidal linear acceleration along the vertical plane. Labyrinthectomized and/or stationary controls showed only sporadically scattered Fos-labeled neurons in the vestibular nuclei, confirming an otolithic origin of c-fos expression. Functionally activated Fos expression in neurons of the medial and spinal vestibular nuclei and group x were identifiable by P7 and those in group y by P9. A small number of Fos-labeled neurons characterized by small soma size were found in the ventral part of lateral vestibular nucleus by P9. Other vestibular-related areas such as prepostitus hypoglossal nucleus, gigantocellular reticular nucleus and locus coeruleus of normal experimental rats showed functionally activated c-fos expression at P7. Neurons in dorsal medial cell column and beta subnucleus of the inferior olive only showed functionally activated c-fos expression by the second postnatal week. These findings revealed a unique critical maturation time for each of the vestibular-related brainstem areas in the recognition of gravity related vertical head orientations. By mapping the three-dimensional distribution of Fos-immunoreactive neurons, we found an even distribution of otolith-related neurons within the spinal vestibular nucleus in groups x and y but a clustered distribution in the middle-lateral-ventral part of the medial vestibular nucleus. Taken together, our findings reveal the developmental profile of neuronal subpopulations within the vertical otolith system, thereby providing an anatomical basis for postnatal coding of gravity-related vertical head movements. PMID- 16706851 TI - Lesions of an avian forebrain nucleus prevent changes in protein kinase C levels associated with deafening-induced vocal plasticity in adult songbirds. AB - We investigated the participation of protein kinase C (PKC) in the regulation of vocal plasticity in songbirds. Deafening of adult Bengalese finches causes initial song alteration, followed by stabilization. In parallel, the expression of PKC beta1 increases transiently 2 weeks after deafening, and then decreases gradually in the robust nucleus of the arcopallium (RA) of Bengalese finches, similar to the pattern observed during developmental song learning. First, we showed that in adult zebra finches, whose songs change more gradually after auditory deprivation than those of Bengalese finches, PKC in RA also increased to an equal degree 2 weeks after deafening, despite the species difference. Second, double-labeling with an anterograde tracer and PKC immunofluorescence revealed that PKC immunoreactivity in RA was detected on the synaptic terminals from a high premotor vocal nucleus (HVC), but not from the lateral magnocellular nucleus of the anterior nidopallium (LMAN). To determine what causes deafening-induced PKC increases, we blocked signals from LMAN, the final output nucleus to RA in the anterior forebrain pathway (AFP), by a unilateral LMAN lesion prior to auditory deprivation of adult Bengalese finches. The PKC immunoreactivity increased in RA of the intact hemisphere; however, in RA on the lesioned side, it was less intense than that of the unlesioned side. Thus, the deafening-induced PKC expression was suppressed by lesioning of LMAN. These results suggest that an output signal from the AFP via LMAN induces the increase in PKC activity on HVC RA synapses that may regulate song plasticity. PMID- 16706852 TI - Short periods of concordant binocular vision prevent the development of deprivation amblyopia. AB - Based in part on deprivation studies, it is generally agreed that the development of vision and of the central visual pathways of higher mammals such as cats and primates is experience-dependent. Past deprivation experiments employed periods of exclusively abnormal early visual input. Because of the absence of any normal visual input, such studies indicate only the extremes to which the visual system can change in response to visually driven activity (i.e. its capabilities) rather than provide insight into the role of early visual input in normal development (i.e. what it actually does). We examined the possibility that certain visual input, i.e. normal concordant binocular vision, may be more efficacious than others with respect to its effects on the developing visual system and on vision. On a daily basis, one type of visual input, i.e. normal binocular experience (BE), was pitted against abnormal (monocular exposure, ME) input in order to see if one was more effective. We show that 2 h of daily normal concordant, but not discordant, BE outweighs or protects against as much as 5 h of daily abnormal input to permit the development of normal grating acuity and alignment accuracy in the two eyes. Further, we show that splitting the period of BE into two 1-h periods straddling the period of ME was ineffective, thereby indicating the 2 h of BE each day must be continuous to protect against the development of amblyopia. PMID- 16706853 TI - The comparable size and overlapping nature of upper limb distal and proximal muscle representations in the human motor cortex. AB - The purpose of this study was to determine the relative size and location of proximal and distal upper limb muscle representations in the human motor cortex. Motor-evoked potentials (MEPs) evoked by transcranial magnetic stimulation were recorded in the proximal muscle anterior deltoid (AD) and in the distal muscles extensor carpi radialis (ECR) and first dorsal interosseus (1DI). The coil was moved in steps of 1 cm along a grid drawn on a tight-fitting polyester cap placed on the subject's head. At each location, four stimuli were delivered at 1.2 times the active motor threshold (AMT), and MEPs averaged in real-time. The peak-to peak amplitude of each muscle's mean MEP was measured at each stimulation site. The area of a muscle's representation was measured by a pixel-counting algorithm. The optimal point of each muscle's areal representation, which corresponds to the locus near which the largest MEPs are obtained, was determined by fitting a 3D Lorentzian function to the data points. The optimal point of distal muscles tended to be situated more laterally along the motor strip than that of proximal muscles. However, there was no statistically significant difference between the size of the areal representations and they overlapped considerably. Additionally, in another five subjects, using a small 45-mm coil placed in a hyper-focal orientation, maps were obtained at a stimulus intensity of 1.1-1.15 times the AMT of the muscle with the lowest threshold, usually the 1DI. Even in this very stringent condition, the mapped representations of the AD, ECR and 1DI overlapped, notwithstanding that sharp demarcations between borders were also apparent. These observations demonstrate that stimulus spread alone does not explain the overlap of muscle representations. These results show that commonly used proximal and distal upper-limb muscles, taken individually, are controlled by motor cortical territories of approximately equal size that significantly overlap despite differences in the location of their optimal points. PMID- 16706855 TI - Paradoxical effects of NPY in the suprachiasmatic nucleus. AB - The circadian clock in the suprachiasmatic nucleus (SCN) is synchronized by the 24 h, light : dark cycle, and is reset by photic and non-photic cues. The acute effects of light in the SCN include the increase of mRNA levels of the circadian clock gene Per1 and a dramatic reduction of pineal melatonin. Neuropeptide Y (NPY), which appears to mediate the phase-resetting effects of non-photic stimuli, prevents the ability of light, and stimuli that mimic light, to phase shift the circadian clock when injected into the SCN. The purpose of the present study was to determine if NPY inhibits the ability of light to suppress pineal melatonin. Surprisingly, NPY injected into the SCN of hamsters mimicked the effects of light by suppressing pineal melatonin levels. To confirm that NPY inhibited the effects of light on the induction of Per1 mRNA levels, Per1 mRNA levels in the SCN were measured in these same animals. NPY significantly reduced Per1 mRNA levels induced by the light pulse. The suppression of melatonin by NPY appears to be mediated by the same subtype of NPY receptors in the SCN that mediate the modulation of phase shifts. Injection of Y5 receptor agonists mimicked the effects of NPY on pineal melatonin, while injection of a Y2 agonist did not. Thus, these data are the first to demonstrate the paradoxical effects of NPY within the SCN. NPY mimics the effects of light on pineal melatonin and inhibits the effects of light on the induction of Per1 mRNA. PMID- 16706854 TI - The light-activated signaling pathway in SCN-projecting rat retinal ganglion cells. AB - In mammals, the master circadian clock resides in the suprachiasmatic nuclei (SCN) of the hypothalamus. The period and phase of the circadian pacemaker are calibrated by direct photic input from retinal ganglion cells (RGCs). SCN projecting RGCs respond to light in the absence of rod- and cone-driven synaptic input, a property for which they are termed intrinsically photosensitive. In SCN projecting RGCs, light activates a nonselective cationic current that displays inward and outward rectification. The goal of the present study was to investigate the identity of the light-activated ion channel and the intracellular signaling pathway leading to its activation. We considered two candidate channels, cyclic nucleotide-gated (CNG) channels and transient receptor potential (TRP) channels, which mediate vertebrate and invertebrate phototransduction, respectively. We report that the intrinsic light response relies upon a G-protein dependent process. Although our data indicate that cyclic nucleotides modulate the signaling pathway, CNG channels do not appear to conduct the light-activated current because (i) cyclic nucleotides in the pipette solution do not activate a conductance or completely block the light response, (ii) CNG channel blockers fail to inhibit the light response, (iii) the effects of internal and external divalent cations are inconsistent with their effects on CNG channels, and (iv) immunohistochemistry reveals no CNG channels in SCN-projecting RGCs. Finally, we show that the pharmacology of the light-activated channel resembles that of some TRPC channel family members; the response is blocked by lanthanides and ruthenium red and SK&F 96365, and is enhanced by flufenamic acid and 1-oleoyl-2-acetyl-sn glycerol. Furthermore, immunohistochemical experiments reveal that TRPC6 is expressed in many RGCs, including those that express melanopsin. PMID- 16706856 TI - Both alpha2 and alpha3 GABAA receptor subtypes mediate the anxiolytic properties of benzodiazepine site ligands in the conditioned emotional response paradigm. AB - Mice with point-mutated alpha2 GABAA receptor subunits (rendering them diazepam insensitive) are resistant to the anxiolytic-like effects of benzodiazepines (BZs) in unconditioned models of anxiety. We investigated the role of the alpha2 GABAA subtype in a model of conditioned anxiety. alpha2(H101R) and wildtype mice were trained in a conditioned emotional response (CER) task, in which lever pressing for food on a variable interval (VI) schedule was suppressed during the presentation of a conditioned stimulus (CS+) that predicted footshock. The ability of diazepam, ethanol and pentobarbital to reduce suppression during the CS+ was interpreted as an anxiolytic response. Diazepam (0, 0.5, 1, 2, 4 and 8 mg/kg) induced a dose-dependent anxiolytic-like effect in wildtype mice. At high doses, diazepam (2, 4 and 8 mg/kg) was sedative in alpha2(H101R) mice. Analysis of the anxiolytic properties of nonsedative diazepam doses (0.5 and 1 mg/kg), showed that alpha2(H101R) mice were resistant to the anxiolytic effects of diazepam. Equivalent anxiolytic properties of pentobarbital (20 mg/kg) and ethanol (1 and 2 g/kg) were seen in both genotypes. These findings confirm the critical importance of the alpha2 GABAA subtype in mediating BZ anxiolysis. However, as a compound, L-838417, with agonist properties at alpha2, alpha3 and alpha5-containing receptors, gave rise to anxiolytic-like activity in alpha2(H101R) mice in the CER test, alpha3-containing GABA receptors are also likely to contribute to anxiolysis. Observations that alpha2(H101R) mice were more active, and displayed a greater suppression of lever pressing in response to fear-conditioned stimuli than wildtype mice, suggests that the alpha2(H101R) mutation may not be behaviourally silent. PMID- 16706857 TI - EEG evidence for the presence of an action observation-execution matching system in children. AB - In the adult human brain, passive observation of actions performed by others activates some of the same cortical areas that are involved in the execution of actions, thereby contributing to action recognition. This mechanism appears to occur through activation of a population of action-coding cells known as mirror neurons (MN). In the adult motor cortex, performing actions and observing human movement reduces the magnitude of the mu (8-13 Hz) rhythm, possibly reflecting MN system activity. Despite the wealth of information available regarding the adult MN system, little is known about its existence in children. Here, we used EEG to probe mu rhythm modulation in 15 children during observation and execution of hand actions. Our data show that mu rhythm attenuation occurs in children under 11 years old during observation of hand movements. Similarly to what has been reported in adults, observation of goal/object-orientated movement produces greater modulation of the mu rhythm than intransitive movement. These data confirm the existence of an observation-execution matching system in the immature human brain and may be of clinical value in the understanding of neurodevelopmental disorders associated with a faulty MN system, such as autism spectrum disorder. PMID- 16706858 TI - ERP and fMRI correlates of endogenous and exogenous focusing of visual-spatial attention. AB - The aim of this study was to investigate the neural correlates of the functional distinction underlying attentional mechanisms of endogenous-sustained and exogenous-transient spatial selection. We recorded event related potentials (ERPs) and used functional magnetic resonance imaging (fMRI) in separate experiments while subjects performed a simple reaction time (RT) to the same visual stimulus displayed to one of several field locations. Endogenous-sustained or exogenous-transient focusing of attention onto target location were obtained by presenting the stimulus in blocks of same-point vs. randomised-point trials, respectively. Same-point stimuli yielded overall faster RT than randomised stimuli, indicating a facilitating effect of endogenous-sustained spatial attention on the perceptual processing of the impending stimulus. Moreover, same point vs. randomised presentations revealed significant increases in the fMRI signal in the bilateral lingual and fusiform gyri as well as in the right calcarine sulcus, in conjunction with a larger amplitude of the posterior P1 component of ERPs, but no modulation of the amplitude of the N1 component. Rather, a larger amplitude of N1 was found in the reverse contrast, randomised minus same-point trials, which revealed increases in the fMRI signal along the posterior left superior frontal sulcus and bilaterally in the superior precuneus. These findings indicate that N1 indexes exogenous orienting of attention and is likely to represent the activity of frontal and parietal components of the attention network involved in eliciting attention changes. In contrast, the effects of those changes, resulting in a modulation of activation in visual occipital areas, are indexed by P1. PMID- 16706859 TI - Motor facilitation during action observation: topographic mapping of the target muscle and influence of the onlooker's posture. AB - Transcranial magnetic stimulation (TMS) studies report that viewing a given action performed by a model activates the neural representation of the onlooker's muscles that are activated during the actual execution of the observed action. Here we sought to determine whether this mirror observation-execution facilitation reflects only muscular specificity or whether it is also influenced by postural congruency between onlooker/model body parts. We recorded motor potentials evoked by single-pulse TMS from the first dorsal interosseous (FDI) and abductor digiti minimi (ADM) muscles during observation of the right index and little finger abduction/adduction movements of models who kept their hands in a palm-down or palm-up position. Moreover, in different experiments observers kept their right hand palm down or palm up. Selective motor facilitation was observed during observation of movements that map the motor function of the targeted muscles, regardless of the posture of the observed hand. Modulation of FDI, however, was obtained only when participants kept their hand palm down; by contrast, modulation of ADM was obtained only when participants kept their hand palm up. Interestingly, electromyographic recordings showed that FDI is mostly active when index abduction/adduction movements are performed in the palm-down position, whereas ADM is mostly active when little finger abduction/adduction movements are performed in the palm-up position. Results show that the influence of the onlooker's hand posture is comparable in action execution and observation, thus indicating a fine-grain functional correspondence between these two processes. PMID- 16706860 TI - Functional and effective connectivity in an fMRI study of an auditory-related task. AB - This study investigates the sets of brain areas that are functionally connected during an auditory goal-directed task. We used a paradigm including a resting state condition and an active condition, which consisted in active listening to the footsteps of walking humans. The regional brain activity was measured using fMRI and the adjusted values of activity in brain regions involved in the task were analysed using both principal component analysis and structural equation modelling. A first set of connected areas includes regions located in Heschl's gyrus, planum temporale, posterior superior temporal sulcus (in the so-called 'social cognition' area), and parietal lobe. This network could be responsible for the perceptual integration of the auditory signal. A second set encompassing frontal regions is related to attentional control. Dorsolateral- and medial prefrontal cortex have mutual negative influences which are similar to those described during a visual goal-directed task [T. Chaminade & P. Fonlupt (2003) Eur. J. Neurosci., 18, 675-679.]. Moreover, the dorsolateral prefrontal cortex (DLPFC) exerts a positive influence on the auditory areas during the task, as well as a strong negative influence on the visual areas. These results show that: (i) the negative influence between the medial and lateral parts of the frontal cortex during a goal-directed task is not dependent on the input modality (visual or auditory), and (ii) the DLPFC activates the pathway of the relevant sensory modality and inhibits the nonrelevant sensory modality pathway. PMID- 16706861 TI - Selective tuning of cortical sound-feature processing by language experience. AB - In 'quantity-languages', such as Japanese or Finnish, sound duration is linguistically relevant. We showed that quantity-language speakers were superior to speakers of a non-quantity language in discriminating the duration of even non speech sounds. In contrast, there was no group difference in the discrimination of sound frequency. This result, obtained both by behavioural and neural indices at attentive and automatic levels of processing, indicates precise feature specific tuning of the auditory-cortex functions by the mother tongue. PMID- 16706862 TI - Developing and evaluating an interactive information skills tutorial. AB - OBJECTIVE: To develop and evaluate a web-based interactive information skills tutorial integrated into the curriculum. To determine whether the tutorial was acceptable to students and explore the use of a skills assessment tool in identifying whether the tutorial improved skills. METHODS: The development of a tutorial on OVID medline to teach transferable information skills. A small cohort study to evaluate students' views on the tutorial and its effects on information skills. RESULTS: Thirteen objective assessments were usable. There was a statistically significant improvement in mean final assessment scores, compared with mean pre-training scores, F(2,14) = 11.493, P = 0.001. Eleven (85%) students had improved their overall information skills. The improvement in overall searching skills was enhanced by referral to the tutorial. CONCLUSIONS: The tutorial was successfully developed and integrated into a Masters programme curriculum. In this setting, it appears to reinforce active learning, and was well received by students, who developed core generic searching skills and demonstrated improved information skills in the short and longer term. Students could use the tutorial for revision and study at a time and place of their choosing. Further evaluation is required to assess the impact of using the tutorial with large groups of students, and as a stand-alone teaching medium. PMID- 16706863 TI - Electronic journal provision in a health-care library: insights from a consultation with NHS workers. AB - OBJECTIVE: This study determines the current awareness journal reading requirements of the users of Stockport National Health Service (NHS) Trust's library. The overlap between requirements and the provision of the NHS Core Content resources, four major electronic journal bundles, and the holdings of North West health libraries is also investigated. METHODS: A survey of both hospital and Primary Care Trust staff was conducted, and respondents were required to provide a list of their favourite journal titles. Each requested title was assigned a subject code, and the impact factor was noted. RESULTS: From 135 survey responses, 217 journal titles were identified and 33 category codes were utilized. There was little overlap between the request list and the NHS Core Content titles, but substantial correspondence existed between the request list and the print holdings of North West health libraries. CONCLUSIONS: Current awareness journal reading requirements will not be met by the Core Content provision alone. Bundles of titles offer value-for-money solutions, but may be at the expense of popular titles. Furthermore, the success of regional document supply schemes may be compromised if large numbers of health-care libraries replace print holdings with similar electronic journal bundles. PMID- 16706864 TI - Fifteen hundred guidelines and growing: the UK database of clinical guidelines. AB - BACKGROUND: The National Library for Health offers a comprehensive searchable database of nationally approved clinical guidelines, called the Guidelines Finder. This resource, commissioned in 2002, is managed and developed by the University of Sheffield Health Sciences Library. METHODS: The authors introduce the historical and political dimension of guidelines and the nature of guidelines as a mechanism to ensure clinical effectiveness in practice. The article then outlines the maintenance and organisation of the Guidelines Finder database itself, the criteria for selection, who publishes guidelines and guideline formats, usage of the Guidelines Finder service and finally looks at some lessons learnt from a local library offering a national service. CONCLUSIONS: Clinical guidelines are central to effective clinical practice at the national, organisational and individual level. The Guidelines Finder is one of the most visited resources within the National Library for Health and is successful in answering information needs related to specific patient care, clinical research, guideline development and education. PMID- 16706865 TI - Improving library services for graduate nurse students in New Zealand. AB - BACKGROUND: Graduate students are expected to have basic information literacy skills. Knowing how to access and select relevant information and then utilize that information is a necessary skill in today's knowledge and information-age world. In the health-care sector, the rise in evidence-based medicine has brought into focus the importance of information literacy skills for nurses. In the case described, information literacy skills are embedded in the curriculum and strengthened in collaboration with library staff. AIMS: To identify graduate student nurses' use of the library, and the library's response to findings. METHODS: A case-study approach was used with both quantitative and qualitative data. An anonymous questionnaire was distributed to all nurses enrolled in graduate courses in the second semester of 2002. Interviews were then undertaken with library staff. RESULTS: The university library services were not used by 43% of graduate nursing students and the library responded by developing a number of initiatives. CONCLUSIONS: Finding a deficit in use of the library by graduate nurses, this paper outlines the response by the library to improving awareness of services, access and providing education in a bid to improve literacy skills. PMID- 16706866 TI - The UK's SMARTAL Project: St MARTin's College health students Access to Learning resources whilst on placement. AB - BACKGROUND: St Martin's College pre-registration health students spend up to half their course on periods of placement, during which times they need to be able to access learning resources. The project aimed to replicate the Health and Education Northumbria Students Access to Learning resources (HENSAL) project. OBJECTIVE: To look at patterns of use and access to learning resources for health students on placement. METHODS: Questionnaires were distributed to students who had recently returned from a placement. Five Practice Placement Facilitators were interviewed to gain their views. RESULTS: There was a 60% return rate (n = 268) for the questionnaire. Most students thought access to learning resources was important yet these were not always available and varied according to placement type. Students still made regular use of the College library while they were on placement but had some difficulties. Students used electronic resources, tending to access them from home. CONCLUSIONS: The results indicated that there were issues which needed to be addressed by the College's Faculty of Health and Social Care, Library and IT services, the local NHS Trust libraries and the local Strategic Health Authority to improve access to and use of learning resources. The findings proved to be very similar to those of the HENSAL project. PMID- 16706867 TI - Demonstrating the positive impact of information support on patient care in primary care: a rapid literature review. AB - AIM: To review the literature on the positive impact of information services, or information resources, on patient care in primary care. OBJECTIVES: To identify and summarize key papers on which librarians might draw in making the case for investment, and to highlight gaps in the research evidence Methodology: A rapid literature review was conducted in the summer of 2005. RESULTS: There is a small body of evidence to demonstrate the positive impact of library and information services on the direct care of patients as well as a beneficial impact on the care of future patients through the application of evidence to multiple patients. CONCLUSIONS: There is relatively limited research evidence of the impact of information, and library services, in primary care, in comparison with hospital settings and the research available is generally reliant on small samples. There is a lack of impact studies conducted with non-clinical staff. The review highlights the value of critical incident technique (CIT). It is possible to gather evidence of the potential for information services to deliver cost savings. PMID- 16706868 TI - Competence in nursing informatics among nursing students and staff at a nursing institute in Sweden. AB - BACKGROUND: Information technology (IT) has slowly been implemented in many health-care organizations (i.e. hospitals). The use of computers has advanced to home care and long-term facilities. Consequently, it is important that nurses are able to demonstrate competence and feel confident in the use of IT. Yet, research has reported that nursing students have varying degrees of competence and experience with IT. AIMS: The purpose of this study was to ascertain nursing students' and nursing educators' self-reported attitudes, skills and experiences using IT. METHODS: Data were collected through a written survey distributed and completed by 247 nursing students (45 of whom were registered nurses who participated in extension courses) and 52 staff members from the nursing faculty at a university in Sweden. RESULTS: Two-thirds of the students and 92% of the nursing educators had access to the Internet from their homes. Of the students, 71% identified their IT skills as being sufficient for their current or future work as a registered nurse. Nurse educators were less confident in the students' ability, reporting that only 29% of nursing students had sufficient IT skills. Comparatively, 48% of the nursing educators believed that they themselves had sufficient IT skills for their role as a nurse educator. CONCLUSIONS: It is important that nursing students have basic IT skills and are familiar with e mail, word-processing and various databases on the World Wide Web. Findings indicate that participants had a deficit in these skills, suggesting the need to integrate IT into the nursing curriculum. PMID- 16706869 TI - Evaluation of the KA24 (Knowledge Access 24) service for health- and social-care staff in London and the south-east of England. Part 1: quantitative. AB - AIMS AND OBJECTIVES: This two-part paper aims to identify the main transferable lessons learned from both the quantitative and qualitative evaluations of the Knowledge Access 24 (KA24) service of online databases and selected full-text journals for health and social care staff in London and the south-east of England. The quantitative evaluation analysed usage rates and user registration with the objective of measuring uptake by previously disadvantaged staff, and to inform the subsequent qualitative survey. METHODS: User and usage data were analysed by type of NHS Trust, by type of user, and by what was being used. The evaluation assessed development in user registration and usage of both databases and journals over a 2-year period. Data were aggregated and analysed both monthly and quarterly. RESULTS: Usage levels increased, but uptake in both the mental health and primary care sectors was comparatively slow. Nurses and allied professionals used the service more than doctors. The increase in usage of full text journals over the usage of databases was marked. CONCLUSIONS: Previously disadvantaged staff used electronic resources. A qualitative survey was needed to identify the main enablers and barriers to uptake. PMID- 16706870 TI - The role of the 'Review of New Foreign Titles on Medical Science' in the library of the Chinese Academy of Medical Sciences. PMID- 16706871 TI - Development of a syllabus for healthcare database training. The experience of libraries serving National Health Service staff in Leeds, UK. PMID- 16706872 TI - Success stories from the library accreditation process in health-care libraries in England. PMID- 16706873 TI - On the shelf?: how unused is your reference collection? PMID- 16706875 TI - Incubating desirable change in our health workforce. PMID- 16706876 TI - Rural eHealth paradox: it's not just geography! PMID- 16706877 TI - Australian issues in the provision of after-hours primary medical care services in rural communities. AB - OBJECTIVE: In 2003 the Rural Doctors Workforce Agency in South Australia (SA) facilitated the 'SA Rural Hospital After Hours Triage Education and Training Program'. It was designed to improve communication between rural general practitioners (GPs) and nurses undertaking after-hours triage, provide training in triage for rural nurses and develop local collaborative after-hours primary medical care models that can be applied in other settings. DESIGN: The program consisted of a series of three workshops. The first workshop provided an opportunity for GPs and nurses to discuss local issues relating to after-hours primary medical care service delivery. This was followed by a one-day workshop on triage for nurses. A follow-up refresher workshop was conducted approximately six months later. SETTING: Twenty-three rural communities in SA. PARTICIPANTS: Rural GPs and nurses working in rural communities. RESULTS: This paper reports on the issues highlighted by clinicians in providing after-hours primary medical care in rural and remote communities. These included community expectations, systems of care, scope of practice, private practice/public hospital interface, and medico legal issues. CONCLUSION: The issues facing after-hours health services in rural communities are not new. There are many opportunities for improvement of systems. A formal program including workshops and training has provided a useful forum to commence service improvements. PMID- 16706878 TI - Far West Area Health Service mental health integration project: model for rural Australia? AB - OBJECTIVE: To see if a new model of service delivery ensures that individuals with a mental illness in rural and remote settings could be assessed, treated and cared for in a more appropriate way. DESIGN: Community mental health teams (CMHTs), general practitioners (GP) and other agencies were provided with clinical and broader support services by consultant psychiatrists from public and private sectors. The occasions of service were logged, audited and relevant provider groups were interviewed. Ethics approval was provided by Human Research Ethics Community of University of New South Wales. SETTING: Far West Area Health Service (FWAHS), remote New South Wales. PARTICIPANTS: An enhanced service was provided for residents, specialist mental health and other healthcare providers. RESULTS: Regular access to psychiatrists for primary and secondary care was achieved in remote communities in FWAHS. 3908 new patients were seen by CMHTs between July 2002 and December 2003 and 380 by visiting psychiatrists between January 2002 and July 2003. Secondary consultation, mentoring and education opportunities were made available by tele-conference and face-to-face for CMHTs and others in FWAHS. GPs and CMHTs in remote settings were satisfied with improved access to psychiatrist care. CONCLUSIONS: This model appears to be sustainable with reasonable levels of funding in FWAHS and may be applicable to other remote contexts. PMID- 16706879 TI - Maternity emergency care: short course in maternity emergencies for remote area health staff with no midwifery qualifications. AB - OBJECTIVE: To improve the maternity emergency skills and knowledge of health service providers, without midwifery qualifications, in remote Australia. DESIGN: A descriptive paper utilising data from a needs assessment and course evaluations used to develop and refine a short course in maternity emergencies. SUBJECTS: Remote area health providers. INTERVENTIONS: A multidisciplinary short course, targeting non-midwives in remote areas, aimed at improving knowledge and skills in detection, management and referral of maternity emergencies. RESULTS: Ten courses have been delivered to 175 participants. Evaluations demonstrate a highly valued program, well received and supported by remote health staff. CONCLUSIONS: The course is an important strategy to improve the maternity services offered to women in remote Australia. PMID- 16706880 TI - Rural pharmacy not delivering on its health promotion potential. AB - OBJECTIVE: To investigate the level and perceived quality of health promotion advice received from rural pharmacists. DESIGN: Self-administered written survey on access to and quality of pharmacy services in rural Western Australia completed by rural residents. SETTING: Rural pharmacy. PARTICIPANTS: Four hundred and eighty-three respondents who regularly used a pharmacy. OUTCOME MEASURES: Items in the survey included frequency of receiving prevention advice and satisfaction ratings on health and pharmacy services. RESULTS: Eighty-eight per cent of respondents had never discussed exercise or diet with their pharmacist and 65% had never discussed preventing health problems. Receiving good prevention advice predicted satisfaction with health services in general but not satisfaction with pharmacy services. CONCLUSION: Pharmacies are being underutilised with respect to their capacity to deliver heath prevention advice and ways to capitalise on this potential need to be investigated. PMID- 16706882 TI - Demographic analysis of currently registered dentists in Western Australia: rural urban divide. PMID- 16706881 TI - Patterns of drug use and associated harms among rural injecting drug users: comparisons with metropolitan injecting drug users. AB - OBJECTIVE: Given the harms associated with injecting drug use to both individuals and community and the paucity of such data from rural areas, the study aimed to compare: patterns of drug use, harms, and service access and utilisation among rural and metropolitan injecting drug users (IDU). DESIGN: Cross-sectional survey, using interviewer-administered structured questionnaire. PARTICIPANTS: One hundred and sixty-four rural and 96 metropolitan IDU from seven different New South Wales Area Health Services, recruited through needle and syringe programs (NSPs), snowballing techniques and advertisement. RESULTS: Age, gender, education and employment were similar for rural and metropolitan participants. Both samples reported use of a range of drugs, but rural participants were less likely than metropolitan participants to report daily heroin use (2% vs 10%), but more likely to report having injected morphine (50% vs 21%) in the last six months. Similar proportions reported using a needle/syringe after another person. Rural participants were less likely to report use of NSPs (36% vs 80%) and reported a number of barriers to NSP access and also to drug treatment services. Rural participants reported a significantly longer period of time between blood-borne virus testing. CONCLUSION: Samples of rural IDU are similar to metropolitan, although report some differences in patterns of drug use. Service provision, including access to new injecting equipment, blood-borne virus testing and drug treatment was found to cause considerable problems for rural IDU. These issues warrant further consideration. PMID- 16706883 TI - Implementing a 'Men's Health Pitstop' in the Riverina, South-west New South Wales. PMID- 16706885 TI - Chronic pain in the rural arena. PMID- 16706884 TI - Total learning environment and the implications for rural student nurse retention. PMID- 16706886 TI - A memorable patient. PMID- 16706890 TI - Nanobacteria--propagating calcifying nanoparticles. AB - Nanobacteria, also known as calcifying nanoparticles (CNP), are controversial infectious agents not matching the current criteria for 'living organism'. Despite the controversy of their classification, they propagate and cause cell death in vitro and are associated or found in many human diseases. Thus, more efforts should be focussed on research on pathogenicity of CNP. PMID- 16706891 TI - Identification and characterization of enterococci from bryndza cheese. AB - AIMS: To identify enterococci isolated from sheep milk cheese--bryndza, and to compare differences in the composition of enterococcal microflora affected by the season, and to evaluate the potential presence of vancomycin resistance and virulence determinants. METHODS AND RESULTS: Bacterial strains were isolated during analysis of bryndza cheese and identified on the genus and species level by phenotypic methods and with commercial biochemical sets. The identification of the species, Enterococcus faecium, Ent. durans and Ent. faecalis, was confirmed by PCR using species-specific primers for ddl genes. PCR was also used for assessment of presence of vanA and vanB genes and virulence determinants gelE, agg and cytolysin genes namely: cylL(L), cylL(S), cylM, cylB and cylA. Among 308 Enterococcus sp. strains, 177 isolates were proved to be Ent. faecium, 59 to be Ent. durans and 41 to be Ent. faecalis. Vancomycin resistance genes vanA and vanB were not detected. Agar plate testing confirmed their absence. Gene gelE, however, was found in 20 Ent. faecalis isolates, but only 13 of them showed gelatinase-positive phenotype. Seven isolates had five cytolysin genes, but none of the isolates exhibited a positive haemolytic phenotype. Four isolates possessed the agg gene. The prevalence of Ent. faecium species was highest in samples from the winter season harvest. CONCLUSIONS: Ent. faecium is the dominant enterococcal species in bryndza cheese and the most prevalent in the winter season product. None of the Enterococcus sp. strains was proved to have vanA or vanB genes and the vancomycin resistance. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first report of enterococcal microflora in bryndza cheese and its evaluation for the presence of vanA and vanB genes as well as virulence determinants. PMID- 16706892 TI - Genetic diversity in Aspergillus parasiticus population from the peanut agroecosystem in Argentina. AB - AIMS: The aims of this work were to identify potential sources of Aspergillus parasiticus inoculum and to evaluate the sclerotial and toxigenic profiles of this species from the peanut agroecosystem in Argentina. Likewise, the genetic diversity of A. parasiticus population was analysed using vegetative compatibility group (VCG) analysis. METHODS AND RESULTS: The A. parasiticus strains were isolated from soil, debris and peanut seeds in Cordoba Province, Argentina. A. parasiticus was recovered from the three sources analysed. Only 11 of 185 A. parasiticus isolates (5.9%) did not produce aflatoxins, while 57% produced sclerotia. Twenty-four VCG were identified from 63 isolates. The VCG diversity index for A. parasiticus, expressed as the number of groups divided by the total number of isolates, was 0.31. In general, there were significant differences among VCG in aflatoxin production. CONCLUSIONS: The presence of aflatoxigenic strains of A. parasiticus in the three substrates suggests that they may be an important source of aflatoxin in Argentina's peanut agroecosystem. The A. parasiticus population shows a low genetic diversity. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study showed data on inoculum distribution, aflatoxin and sclerotia production and genetic diversity in an A. parasiticus population isolated from the peanut agroecosystem in Argentina. PMID- 16706893 TI - Effects of butyrate precursors on electron relocation when methanogenesis is inhibited in ruminal mixed cultures. AB - AIMS: Inhibition of ruminal methanogenesis often causes accumulation of H(2), formate and ethanol, which are not energy substrates for ruminants. It was hypothesized that the addition of butyrate precursors would avoid the formation of these products and relocate electrons into butyrate. METHODS AND RESULTS: In four ruminal 24-h incubations, two inhibitors of methanogenesis, each at three different initial concentrations (0, 2 or 4 mmol l(-1) for propynoic acid, and 0, 4 or 8 mmol l(-1) for ethyl 2-butynoate), were combined with two butyrate precursors at two different initial concentrations (0 or 4 mmol l(-1) for crotonic acid or 3-butenoic acid). Ground lucerne hay was the substrate. Propynoic acid at 4 mmol l(-1) decreased CH(4) formation by more than two-thirds. Ethyl 2-butynoate at 8 mmol l(-1) suppressed methanogenesis by more than 90%. Butyrate precursors generally did not decrease the accumulation of H(2) and formate or ethanol production. CONCLUSIONS: Butyrate precursors were ineffective as electron acceptors because they were not completely converted to butyrate and were also metabolized through other pathways. SIGNIFICANCE AND IMPACT OF THE STUDY: Effectiveness of butyrate precursors may be improved by adding them to the fermentation continuously or by enhancing the kinetics of their conversion into butyrate. PMID- 16706894 TI - Effect of the cryopreservation conditions on the viability of the rumen ciliate Diploplastron (Metadinium) affine. AB - AIMS: To study the viability of Diploplastron (Metadinium) affine after its cryopreservation at two cooling rates, and the effect of procedure conditions on viability. METHODS AND RESULTS: There were differences in viability between cooling rates (1 and 4 degrees C min(-1)) at 15 or 5 degrees C, but not after thawing. When the equilibrium temperature (25 or 5 degrees C), the cryopreservant (glycerol or dimethyl sulfoxide [DMSO]) and the use of membrane protector were tested, there were no differences caused by the cryopreservant or the membrane protector. However, the equilibrium at 25 degrees C increased the viability (P = 0.005) compared with 5 degrees C. CONCLUSIONS: Viability after thawing was 0.10 0.19. Adding the cryopreservant (either glycerol or DMSO) at 25 degrees C instead of 5 degrees C improves viability of D. affine after thawing. SIGNIFICANCE AND IMPACT OF THE STUDY: Conditions of cryopreservation are largely dependent on the species of rumen protozoa. Number of viable cells after thawing would indicate the possibility of culture recovery for D. affine. PMID- 16706895 TI - A modified Vibrio harveyi mutagenicity assay based on bioluminescence induction. AB - AIMS: Mutagenic pollution of natural environment is currently one of the most serious ecological problems. Therefore, rapid detection of the presence of mutagens is a very important issue. Although many mutagenicity assays have already been described, only a few are suitable for testing samples from natural environment. One of such assays is a microbiological mutagenicity test based on genetically modified Vibrio harveyi strains. The aim of this work was to modify and improve the V. harveyi assay. METHODS AND RESULTS: A series of V. harveyi dark and dim mutants were tested for reversion of their phenotype towards efficient light emission in response to incubation with known mutagens. Luminescence of the A16 strain (luxE mutant) increased significantly after a few hours of such a treatment with various mutagenic agents, revealing a dose response correlation. Sensitivity of the assay has been determined for different mutagens. CONCLUSIONS: The luminescence-based V. harveyi mutagenicity assay is rapid, sensitive and reveals a dose-response correlation for various mutagens. SIGNIFICANCE AND IMPACT OF THE STUDY: The assay developed in this study is a potentially useful tool in studies on mutagenic pollution of environment, especially marine water. PMID- 16706896 TI - Prevalence of Escherichia coli O157:H7 and Salmonella in beef steers consuming different forage diets. AB - AIMS: To compare the prevalence of faecal shedding of Escherichia coli O157:H7 and Salmonella in growing beef cattle consuming various forages. METHODS AND RESULTS: In Experiment I, faecal samples were collected from steers grazing either endophyte-infected (E+) tall fescue or common bermudagrass (CB). Steers grazing E+ tall fescue were confined to a dry-lot pen and fed CB hay ad libitum for 10 days. In Exp. II, faecal samples were collected from steers grazing either E+ or novel endophyte-infected (NE) tall fescue and treated with one of two anthelmintics: ivermectin (I) or fenbendazole (F). In Exp. I, prevalence of E. coli O157:H7 was less in E+ tall fescue steers fed CB hay than steers grazing CB. More I-treated steers shed Salmonella than F-treated steers at 42-day postanthelmintic treatment but shedding of Salmonella was similar between anthelmintics at day 63 in Exp. II. CONCLUSIONS: Faecal shedding of pathogenic bacteria was not affected by grazing E+ tall fescue. Alterations of forage diets may influence the prevalence of E. coli O157:H7, and anthelmintic treatment could affect faecal shedding of Salmonella. SIGNIFICANCE AND IMPACT OF THE STUDY: Knowledge of factors that influence shedding of pathogenic bacteria in cattle is necessary to develop on-farm intervention strategies aimed at reducing pathogen shedding. PMID- 16706897 TI - Purification and characterization of an extracellular serine protease from the nematode-trapping fungus Dactylella shizishanna. AB - AIMS: To evaluate the production of an extracellular serine protease by Dactylella shizishanna and its potential as a pathogenesis factor. METHODS AND RESULTS: An extracellular alkaline serine protease (Ds1) was purified and characterized from the nematode-trapping fungus D. shizishanna using cation exchange chromatography and hydrophobic interaction chromatography. The molecular mass of the protease was approximately 35 kDa estimated by SDS-PAGE. The optimum activity of Ds1 was at pH 10 and 55 degrees C (over 30 min). The purified protease could degrade purified cuticle of Penagrellus redivivus and a broad range of protein substrates. The purified protease was highly sensitive to phenylmethyl sulfonyl fluoride (PMSF) (0.1 mmol l(-1)), indicating it belonged to the serine protease family. The N-terminal amino acid residues of Ds1 are AEQTDSTWGL and showed a high homology with Aozl and PII, two serine proteases purified from the nematode-trapping fungus Arthrobotrys oligospora. CONCLUSIONS: Nematicidal activity of D. shizishanna was partly related to its ability to produce extracellular serine protease. SIGNIFICANCE AND IMPACT OF THE STUDY: In this report, we purified a new serine protease from D. shizishanna and provided a good foundation for future research on infection mechanism. PMID- 16706898 TI - Manufacturing of fermented goat milk with a mixed starter culture of Bifidobacterium animalis and Lactobacillus acidophilus in a controlled bioreactor. AB - AIMS: This work was undertaken to study the feasibility and the characteristics of a fermented product made of goat milk, using a mixed starter culture of Bifidobacterium animalis and Lactobacillus acidophilus under controlled conditions, and to determine their survival in the fermented milk during refrigerated storage. METHODS AND RESULTS: Goat milk was inoculated with Lact. acidophilus and Bif. animalis mixed starter, fermented in a glass bioreactor with controlled temperature (37 degrees C) and anaerobiosis, and monitored for growth and acidification. The fermented milk was then stored for 10 days under refrigeration, and monitored daily for starter microflora survival and pH changes. Lact. acidophilus viable counts reached a maximum of 7.1 x 10(8) colony forming units (CFU) ml(-1), and Bif. animalis a maximum of 6.3 x 10(7) CFU ml(-1) by 20 h of fermentation. During refrigerated storage, both strains exhibited a good survival, with viable numbers remaining essentially constant throughout the experiment, whereas the pH of the fermented milk dropped slightly. CONCLUSIONS: Mixed cultures of Bif. animalis and Lact. acidophilus may be used to produce fermented goat milk with high counts of both probiotic strains. SIGNIFICANCE AND IMPACT OF THE STUDY: Goat milk fermented with Bif. animalis and Lact. acidophilus can be manufactured as an alternative probiotic dairy product. PMID- 16706899 TI - The use of (GTG)5 oligonucleotide as an RAPD primer to type Campylobacter concisus. AB - AIM: DNA fingerprinting using (GTG)(5) oligonucleotide as a primer in a random amplified polymorphic DNA (RAPD) assay was assessed by typing isolates of Campylobacter concisus strains, collected over a period of 8 years. METHODS AND RESULTS: RAPD analysis using the (GTG)(5) oligonucleotide as a primer was used to type 100 isolates of C. concisus comprising mostly isolates from children with diarrhoea. Using this method, 86% of the isolates were found to be genotypically diverse. Of these heterogeneous isolates, 25 of the strains were also shown to be genetically distinct in a previous study using pulsed field gel electrophoresis. The remaining isolates (14) could be classified into five profile groups based on the DNA fingerprinting patterns. The assay successfully identified epidemiologically linked strains from the unrelated genetically diverse pool of strains. CONCLUSIONS: Laboratory RADP typing using the (GTG)(5) primer proved to be useful in distinguishing related strains of C. concisus from a large pool of unrelated strains of this organism. SIGNIFICANCE AND IMPACT OF THE STUDY: RAPD typing using (GTG)(5) is a simple method that could be used to investigate the epidemiology of C. concisus. The results suggest that homologous lineages of C. concisus may exist within an otherwise heterogeneous species complex. However, these data need to be confirmed using a more robust typing method. PMID- 16706900 TI - Selective growth of Staphylococcus aureus from flushed dairy manure wastewater using acriflavine-supplemented mannitol salt agar. AB - AIMS: To investigate the use of mannitol salt agar (MSA) supplemented with acriflavine for selective growth and quantification of Staphylococcus aureus from flushed dairy manure wastewater (FDMW). METHODS AND RESULTS: Minimal inhibitory concentrations of acriflavine in MSA were determined by comparing the growth of S. aureus subsp. aureus (ATCC 33591) and Staphylococcus epidermidis (ATCC 155) in pure culture. Acriflavine concentrations of 1.3, 1.4 and 1.5 mg l(-1) reduced CFU of S. epidermidis by 43%, 55% and 87%, respectively, while CFU of S. aureus subsp. aureus were only reduced by 15%, 20% and 26% at the respective concentrations of acriflavine. MSA supplemented with 1.5 mg l(-1) acriflavine was tested for selective growth of indigenous S. aureus from three grab samples of FDMW. Acriflavine concentrations of 1.5 mg l(-1) reduced background flora without significantly reducing (P < 0.05) indigenous S. aureus counts. CONCLUSIONS: Acriflavine-supplemented MSA provides an effective media for selective growth and quantification of indigenous S. aureus from FDMW in the presence of high levels of background microflora. SIGNIFICANCE AND IMPACT OF THE STUDY: S. aureus is implicated for mastitis infections in dairy cows. Therefore, a reliable means for monitoring and detecting the organism in FDMW provides a tool for measuring the effectiveness of treatment for reducing S. aureus levels and implementing flushwater recycling without affecting herd health. PMID- 16706901 TI - Laccase activity from the fungus Trametes hirsuta using an air-lift bioreactor. AB - AIM: To produce high laccase activities from the white-rot fungus Trametes hirsuta in an in-house air-lift bioreactor (ALB). METHODS AND RESULTS: Trametes hirsuta was grown in a 6-l ALB. A fed-batch strategy with glycerol as an addition resulted in maximum laccase activity of 19,400 U l(-1), which was the highest reported from the fungus. CONCLUSION: The ALB configuration with additional glycerol resulted in high laccase activities. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides useful information on how to produce high concentrations of laccase. PMID- 16706902 TI - Effect of reduced pH on inorganic polyphosphate accumulation by Burkholderia cepacia complex isolates. AB - AIMS: Burkholderia cepacia complex (Bcc) isolates causing pulmonary infection in cystic fibrosis (CF) patients grow within an acidic environment in the lung. As exposure to acid pH has been shown to increase intracellular inorganic polyphosphate (polyP) formation in some bacteria, we investigated the inter relationship between acidic pH and polyP accumulation in Bcc isolates. METHODS AND RESULTS: The formation of polyP by one Burkholderia cenocepacia clinical isolate was initially examined at a range of pH values by measuring total intracellular polyP accumulation and phosphate uptake. The pattern of polyP accumulation corresponded with the pattern of phosphate uptake with the maximum for both occurring at pH 5.5. Phosphate uptake and formation of polyP by this isolate was further determined over 48 h at pH 5.5, 6.5 and 7.5; formation of polyP was maximal at pH 5.5 at all time points studied. Sixteen of 17 additional clinical and environmental Bcc isolates examined also exhibited maximum phosphate uptake at pH 5.5. CONCLUSIONS: Both clinical and environmental Bcc isolates, of five genomovars, show enhanced formation of polyP in an acidic environment. Given both the speculated role of polyP in pathogenesis, cell signalling and biofilm formation and the acidic nature of the CF lung, this may be of considerable clinical importance. SIGNIFICANCE AND IMPACT OF THE STUDY: Growth of Bcc in an acidic environment, such as that found in the lungs of CF patients may be influenced in part by polyP accumulation. PMID- 16706903 TI - A multiplex real-time PCR assay for detection of Xanthomonas campestris from brassicas. AB - AIMS: To develop a sensitive real-time PCR-based protocol for the detection of Xanthomonas campestris pathovars from Brassica seed. METHODS AND RESULTS: A 5' nuclease real-time PCR assay was developed to screen Brassica spp. seed for the presence of X. campestris pathovars that cause black rot. The assay amplifies a 78-bp segment of the X. campestris hrpF gene and a 100-bp segment of the Brassica spp. 18S-25S internal transcribed spacer region. The Brassica spp. target provides an internal control for the amplification process to prevent false negatives that may arise from inhibitors that are often present in extracts from plant material. Whilst the primers were compatible with SYBR Green I assays, the use of fluorescently labelled probes in a 5' nuclease assay afforded greatest sensitivity and specificity. Seed batches carrying one artificially infected seed among 10,000 were readily detected using the assay. The multiplex real-time PCR assay permitted the rapid detection of pathogenic strains of X. campestris from bacterial colonies, Brassica seed and plants. CONCLUSIONS: Strains of X. campestris pathogenic to brassicas were readily detected from seed via a multiplex 5' nuclease real-time PCR assay. The real-time assay offers an improvement in sensitivity and a reduced turn-around time over the conventional multiplex PCR. SIGNIFICANCE AND IMPACT OF THE STUDY: Real-time PCR can be used to rapidly screen Brassica spp. seed batches for the presence of X. campestris pathovars. This assay provides a means for growers and the seed industry to be aware of the black rot status of their planting material, so that they may more effectively employ disease control measures or seed disinfection. PMID- 16706904 TI - Transformation of the entomopathogenic fungus Paecilomyces fumosoroseus with Agrobacterium tumefaciens. AB - AIMS: To test the suitability of the Agrobacterium tumefaciens-mediated transformation (AMT) method with Paecilomyces fumosoroseus, a fungal pathogen that causes diseases in a wide range of insects including whiteflies. METHODS AND RESULTS: Conidia of P. fumosoroseus were successfully transformed to hygromycin B resistance using the hph gene of Escherichia coli as the selectable marker. Transformation frequencies were 58.3 +/- 18.5, 98.3 +/- 24.8 and 169.7 +/- 35.5 (+/-SEM) transformants per 10(5), 10(6) and 10(7) target conidia respectively. After confirmation by PCR, transformants were subjected to Southern analysis, and the results revealed that 45% (four of nine) of the transformants contained single-copy integration of the T-DNA. CONCLUSIONS: In our AMT system, we efficiently transformed conidia of P. fumosoroseus. The employment of this method circumvents time-consuming protoplast preparation and allows the isolation of transformants containing single-copy integration of the T-DNA. SIGNIFICANCE AND IMPACT OF THE STUDY: Considering the efficiency of Ag. tumefaciens-mediated transformation, this method represents a useful tool for insertional mutagenesis to characterize genes that are important for the pathogenicity of P. fumosoroseus. PMID- 16706905 TI - Inhibition of bacterial quorum sensing by vanilla extract. AB - AIMS: The purpose of this study was to search for a novel quorum sensing inhibitor and analyse its inhibitory activity. METHODS AND RESULTS: Quorum sensing inhibition was monitored using the Tn-5 mutant, Chromobacterium violaceum CV026. Vanilla beans (Vanilla planifolia Andrews) were extracted using 75% (v/v) aqueous methanol and added to C. violaceum CV026 cultures. Inhibitory activity was measured by quantifying violacein production using a spectrophotometer. The results have revealed that vanilla extract significantly reduced violacein production in a concentration-dependent manner, indicating inhibition of quorum sensing. CONCLUSIONS: Vanilla, a widely used spice and flavour, can inhibit bacterial quorum sensing. SIGNIFICANCE AND IMPACT OF THE STUDY: The results suggest that the intake of vanilla-containing food materials might promote human health by inhibiting quorum sensing and preventing bacterial pathogenesis. Further studies are required to isolate specific substances from vanilla extract acting as quorum sensing inhibitors. PMID- 16706906 TI - Role of colanic acid exopolysaccharide in the survival of enterohaemorrhagic Escherichia coli O157:H7 in simulated gastrointestinal fluids. AB - AIM: This study evaluated the production of colanic acid (CA) exopolysaccharide (EPS) by Escherichia coli O157:H7 in relation to the pathogen's ability to survive under acidic conditions simulating the environment in the human gastrointestinal tract. METHODS AND RESULTS: Escherichia coli O157:H7 W6-13 and its CA-deficient mutant M4020 were examined for their resistance to bile salts, and their ability to survive in simulated gastric fluid containing pepsin (pH 2.0) and simulated intestinal fluid containing pancreatin (pH 8.0). The effect of acid adaptation at pH 5.5 on the survival of E. coli O157:H7 in simulated gastric fluid was also determined. The results indicated that the survivability of M4020, under conditions simulating the environment in the human gastrointestinal tract, reduced more drastically than the viability of W6-13. The presence of bile salts had a slight effect on both types of E. coli O157:H7 cells. The loss of CA did not change the ability of M4020 to respond to acid adaptation. CONCLUSION: The EPS CA may serve as a protective barrier to E. coli O157:H7 for its survival in the human gastrointestinal tract. SIGNIFICANCE AND IMPACT OF THE STUDY: The study contributes to a better understanding of the EPS affecting the ability of E. coli O157:H7 to combat acid stress. PMID- 16706907 TI - Drug resistance and random amplified polymorphic DNA analysis of Photobacterium damselae ssp. piscicida isolates from cultured Seriola (yellowtail, amberjack and kingfish) in Japan. AB - AIMS: To investigate the antimicrobial susceptibility and genetic characteristics of Photobacterium damselae ssp. piscicida isolates obtained from cultured Seriola in Japan. METHODS AND RESULTS: Minimal inhibitory concentrations of 14 antimicrobials for 74 isolates from Seriola in Japan in 2002 were determined. Isolates showed high frequencies of resistance to sulfamonomethoxine (SMMX) (97.3%), oxytetracycline (OTC) (77.0%), flumequine (FMQ) (77.0%), chloramphenicol (CP) (75.7%), kanamycin (KM) (63.5%) and oxolinic acid (OA) (62.0%), but low to ampicillin (ABPC) (2.8%). All isolates were susceptible to bicozamycin (BCM), fosfomycin (FOM) and florfenicol (FF). Of these isolates, 45 (60.8%) showed same resistance pattern (SMMX-OTC-FMQ-OA-CP-KM). In random amplified polymorphic DNA (RAPD) analysis, no difference was observed among our 74 field isolates and ATCC51736 isolated from Seriola in 1974 in Japan, but different from ATCC 17911 isolated from white perch in USA. CONCLUSIONS: FF, BCM, FOM and ABPC were useful antimicrobials for treating pseudotuberculosis. However, the frequency of multidrug resistance was high. RAPD analysis showed homogeneity of isolates from Seriola in Japan. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates that some antimicrobials were still useful for treating pseudotuberculosis and that P. damselae ssp. piscicida strains of same origin might have spread among Seriola in Japan since 1974. PMID- 16706909 TI - Environmentally induced migration: the importance of food. AB - The decision to migrate or not is regarded as genetically controlled for many invertebrate and vertebrate taxa. Here, we show that the environment influences this decision. By reciprocally transplanting brown trout (Salmo trutta L.) between two sections in a river, we show that both migratory and non-migratory behaviour can be environmentally induced; migratory behaviour developed in a river section with high brown trout densities and low specific growth rates, whereas non-migratory behaviour developed in a section with low brown trout densities and high specific growth rates. In a laboratory experiment, we tested the effect of food availability on the development of migratory and non-migratory body morphologies and found that most brown trout became migrants when food levels were low but fewer did so at high food levels. Thus, the decision to migrate seems to be a plastic response, influenced by growth opportunities. PMID- 16706910 TI - Dispersal frequency affects local biomass production by controlling local diversity. AB - Dispersal is a major factor regulating the number of coexisting species, but the relationship between species diversity and ecosystem processes has mainly been analysed for communities closed to dispersal. We experimentally investigated how initial local diversity and dispersal frequency affect local diversity and biomass production in open benthic microalgal metacommunities. Final local species richness and local biomass production were strongly influenced by dispersal frequency but not by initial local diversity. Both final local richness and final local biomass showed a hump-shaped pattern with increasing dispersal frequency, with a maximum at intermediate dispersal frequencies. Consequently, final local biomass increased linearly with increasing final richness. We conclude that the general relationship between richness and ecosystem functioning remains valid in open systems, but the maintenance of ecosystem processes significantly depends on the effects of dispersal on species richness and local interactions. PMID- 16706911 TI - A null model of temporal trends in biological invasion records. AB - Biological invasions are a growing aspect of global biodiversity change. In many regions, introduced species richness increases supralinearly over time. This does not, however, necessarily indicate increasing introduction rates or invasion success. We develop a simple null model to identify the expected trend in invasion records over time. For constant introduction rates and success, the expected trend is exponentially increasing. Model extensions with varying introduction rate and success can also generate exponential distributions. We then analyse temporal trends in aquatic, marine and terrestrial invasion records. Most data sets support an exponential distribution (15/16) and the null invasion model (12/16). Thus, our model shows that no change in introduction rate or success need be invoked to explain the majority of observed trends. Further, an exponential trend does not necessarily indicate increasing invasion success or 'invasional meltdown', and a saturating trend does not necessarily indicate decreasing success or biotic resistance. PMID- 16706912 TI - Temperature-dependence of biomass accumulation rates during secondary succession. AB - Rates of ecosystem recovery following disturbance affect many ecological processes, including carbon cycling in the biosphere. Here, we present a model that predicts the temperature dependence of the biomass accumulation rate following disturbances in forests. Model predictions are derived based on allometric and biochemical principles that govern plant energetics and are tested using a global database of 91 studies of secondary succession compiled from the literature. The rate of biomass accumulation during secondary succession increases with average growing season temperature as predicted based on the biochemical kinetics of photosynthesis in chloroplasts. In addition, the rate of biomass accumulation is greater in angiosperm-dominated communities than in gymnosperm-dominated ones and greater in plantations than in naturally regenerating stands. By linking the temperature-dependence of photosynthesis to the rate of whole-ecosystem biomass accumulation during secondary succession, our model and results provide one example of how emergent, ecosystem-level rate processes can be predicted based on the kinetics of individual metabolic rate. PMID- 16706913 TI - Multivariate dispersion as a measure of beta diversity. AB - Beta diversity can be defined as the variability in species composition among sampling units for a given area. We propose that it can be measured as the average dissimilarity from individual observation units to their group centroid in multivariate space, using an appropriate dissimilarity measure. Differences in beta diversity among different areas or groups of samples can be tested using this approach. The choice of transformation and dissimilarity measure has important consequences for interpreting results. For kelp holdfast assemblages from New Zealand, variation in species composition was greater in smaller holdfasts, while variation in relative abundances was greater in larger holdasts. Variation in community structure of Norwegian continental shelf macrobenthic fauna increased with increases in environmental heterogeneity, regardless of the measure used. We propose a new dissimilarity measure which allows the relative weight placed on changes in composition vs. abundance to be specified explicitly. PMID- 16706914 TI - Ecological and evolutionary principles in immunology. AB - Experimental immunology has given rise to detailed insights into how immune cells react to infectious agents and fight pathogens. At the same time, however, the interplay between infectious agents and immune responses can be viewed as an ecological system in vivo. This is characterized by complex interactions between species of immune cells and populations of pathogens. This review discusses how an understanding of the immune system can be aided by the application of ecological and evolutionary principles: competition, predation, and the evolution of viruses in vivo. These concepts can shed light onto important immunological concepts such as the correlates of efficient virus control, immunodominance, the relationship between viral evolution and the development of pathology, as well as the ability of the immune system to control immunosuppressive infections. PMID- 16706915 TI - Transmission assumptions generate conflicting predictions in host-vector disease models: a case study in West Nile virus. AB - This review synthesizes the conflicting outbreak predictions generated by different biological assumptions in host-vector disease models. It is motivated by the North American outbreak of West Nile virus, an emerging infectious disease that has prompted at least five dynamical modelling studies. Mathematical models have long proven successful in investigating the dynamics and control of infectious disease systems. The underlying assumptions in these epidemiological models determine their mathematical structure, and therefore influence their predictions. A crucial assumption is the host-vector interaction encapsulated in the disease-transmission term, and a key prediction is the basic reproduction number, R(0). We connect these two model elements by demonstrating how the choice of transmission term qualitatively and quantitatively alters R(0) and therefore alters predicted disease dynamics and control implications. Whereas some transmission terms predict that reducing the host population will reduce disease outbreaks, others predict that this will exacerbate infection risk. These conflicting predictions are reconciled by understanding that different transmission terms apply biologically only at certain population densities, outside which they can generate erroneous predictions. For West Nile virus, R(0) estimates for six common North American bird species indicate that all would be effective outbreak hosts. PMID- 16706916 TI - Biotic interactions and plant invasions. AB - Introduced plant populations lose interactions with enemies, mutualists and competitors from their native ranges, and gain interactions with new species, under new abiotic conditions. From a biogeographical perspective, differences in the assemblage of interacting species, as well as in abiotic conditions, may explain the demographic success of the introduced plant populations relative to conspecifics in their native range. Within invaded communities, the new interactions and conditions experienced by the invader may influence both its demographic success and its effects on native biodiversity. Here, we examine indirect effects involving enemies, mutualists and competitors of introduced plants, and effects of abiotic conditions on biotic interactions. We then synthesize ideas building on Darwin's idea that the kinds of new interactions gained by an introduced population will depend on its relatedness to native populations. This yields a heuristic framework to explain how biotic interactions and abiotic conditions influence invader success. We conclude that species introductions generally alter plants' interactions with enemies, mutualists and competitors, and that there is increasing evidence that these altered interactions jointly influence the success of introduced populations. PMID- 16706917 TI - Functional diversity: back to basics and looking forward. AB - Functional diversity is a component of biodiversity that generally concerns the range of things that organisms do in communities and ecosystems. Here, we review how functional diversity can explain and predict the impact of organisms on ecosystems and thereby provide a mechanistic link between the two. Critical points in developing predictive measures of functional diversity are the choice of functional traits with which organisms are distinguished, how the diversity of that trait information is summarized into a measure of functional diversity, and that the measures of functional diversity are validated through quantitative analyses and experimental tests. There is a vast amount of trait information available for plant species and a substantial amount for animals. Choosing which traits to include in a particular measure of functional diversity will depend on the specific aims of a particular study. Quantitative methods for choosing traits and for assigning weighting to traits are being developed, but need much more work before we can be confident about trait choice. The number of ways of measuring functional diversity is growing rapidly. We divide them into four main groups. The first, the number of functional groups or types, has significant problems and researchers are more frequently using measures that do not require species to be grouped. Of these, some measure diversity by summarizing distances between species in trait space, some by estimating the size of the dendrogram required to describe the difference, and some include information about species' abundances. We show some new and important differences between these, as well as what they indicate about the responses of assemblages to loss of individuals. There is good experimental and analytical evidence that functional diversity can provide a link between organisms and ecosystems but greater validation of measures is required. We suggest that non-significant results have a range of alternate explanations that do not necessarily contradict positive effects of functional diversity. Finally, we suggest areas for development of techniques used to measure functional diversity, highlight some exciting questions that are being addressed using ideas about functional diversity, and suggest some directions for novel research. PMID- 16706918 TI - Prospecting for pig single nucleotide polymorphisms in the human genome: have we struck gold? AB - Gene-to-gene variation in the frequency of single nucleotide polymorphisms (SNPs) has been observed in humans, mice, rats, primates and pigs, but a relationship across species in this variation has not been described. Here, the frequency of porcine coding SNPs (cSNPs) identified by in silico methods, and the frequency of murine cSNPs, were compared with the frequency of human cSNPs across homologous genes. From 150,000 porcine expressed sequence tag (EST) sequences, a total of 452 SNP-containing sequence clusters were found, totalling 1394 putative SNPs. All the clustered porcine EST annotations and SNP data have been made publicly available at http://sputnik.btk.fi/project?name=swine. Human and murine cSNPs were identified from dbSNP and were characterized as either validated or total number of cSNPs (validated plus non-validated) for comparison purposes. The correlation between in silico pig cSNP and validated human cSNP densities was found to be 0.77 (p < 0.00001) for a set of 25 homologous genes, while a correlation of 0.48 (p < 0.0005) was found for a primarily random sample of 50 homologous human and mouse genes. This is the first evidence of conserved gene-to gene variability in cSNP frequency across species and indicates that site directed screening of porcine genes that are homologous to cSNP-rich human genes may rapidly advance cSNP discovery in pigs. PMID- 16706919 TI - Can low densities of carnivores result in genetic depletion? An investigation within French polecat populations. AB - Carnivores as top predators are commonly found in relatively low densities even in optimal habitats. Despite a lack of empirical studies, it could be suspected that long-term low density could result in genetic depletion. The genetic structure of European polecat Mustela putorius natural populations was investigated by allozyme electrophoresis in five distinct areas. Density estimates significantly differed among sites from 0.17 to 0.83 individuals/km2 with an average of 0.56 individuals per km2, resulting in a scattered distribution. Genetic structure varied among distinct populations both in number of polymorphic loci and heterozygosity. Polecats from Brittany revealed a very low observed heterozygosity (H(O) = 0.028) whereas mean heterozygosity reached H(O) = 0.072 in Briere. That the lowest heterozygosity levels and highest inbreeding coefficient F(IS) were significantly associated with the lowest densities suggests that low densities may affect populations of carnivores. Both the loss of polymorphic loci and the reduction in heterozygosity rates suggest a density-dependent effect and population density can be arguably regarded as a factor affecting genetic diversity in top carnivores. PMID- 16706920 TI - Epistatic kinship a new measure of genetic diversity for short-term phylogenetic structures--theoretical investigations. AB - The epistatic kinship describes the probability that chromosomal segments of length x in Morgan are identical by descent. It is an extension from the single locus consideration of the kinship coefficient to chromosomal segments. The parameter reflects the number of meioses separating individuals or populations. Hence it is suggested as a measure to quantify the genetic distance of subpopulations that have been separated only few generations ago. Algorithms for the epistatic kinship and the extension of the rules to set up the rectangular relationship matrix are presented. The properties of the epistatic kinship based on pedigree information were investigated theoretically. Pedigree data are often missing for small livestock populations. Therefore, an approach to estimate epistatic kinship based on molecular marker data are suggested. For the epistatic kinship based on marker information haplotypes are relevant. An easy and fast method that derives haplotypes and the respective frequencies without pedigree information was derived based on sampled full-sib pairs. Different parameters of the sampling scheme were tested in a simulation study. The power of the method decreases with increasing segment length and with increasing number of segments genotyped. Further, it is shown that the efficiency of the approach is influenced by the number of animals genotyped and the polymorphism of the markers. It is discussed that the suggested method has a considerable potential to allow a phylogenetic differentiation between close populations, where small sample size can be balanced by the number, the length, and the degree of polymorphism of the chromosome segments considered. PMID- 16706921 TI - Evaluation of genetic diversity in Japanese Brown cattle population by pedigree analysis. AB - The Japanese Brown is the second most common domestic beef breed in Japan. However, nowadays this breed is facing reduction in numbers because of pressure from a profitable domestic breed. This breed is uniformly characterized by its brown coat colour, but is comprised of two isolated sub-breeds, Kumamoto and Kouchi, each possessing a different gene pool. Pedigree analyses were carried out for the two sub-breeds using the pedigree records of animals born from 1970 to 2000. The effective population size has been found to be consistently reducing during the last three decades in both sub-breeds. The current effective sizes were estimated to be 25.5 and 6.0 for the Kumamoto and Kouchi sub-breeds, respectively. The estimate of the effective number of founders (N(ef)) in the Kumamoto sub-breed decreased from 152.1 to 74.4; that of non-founders (N(enf)), from 41.7 to 5.3; and that of founder genome equivalents (N(ge)), from 32.7 to 4.9. The corresponding changes in the Kouchi sub-breed were from 108.2 to 79.4, 16.2 to 4.1, and 14.1 to 3.9. Increasing differences between the two genetic diversity indices in the sub-breeds indicate that the greater part of the reduction of genetic diversity can be attributed to genetic drift that accumulated in the non-founder generations. A comparison with published estimates for several cattle breeds suggests the extremely limited genetic diversity of Japanese Brown. In addition to the avoidance of further reduction of genetic diversity, it will be important to counteract the process of breed decline by establishing a production system to efficiently utilize the unique characteristics of this breed and by developing links between the breed and products with market value. PMID- 16706923 TI - Genetic and permanent environmental variability of twinning in Thoroughbred horses estimated via three threshold models. AB - Twinning is a serious problem in Thoroughbred horses, as most of the multiple pregnancies are miscarried and live foals present lower performance values. The aim of this study was to estimate the heritability and repeatability of twinning in Thoroughbred horses. A total of 12648 pregnancies of 2033 Thoroughbred mares born between 1929 and 1994 in 11 studs were included. The analysis was performed by the use of AI-REML algorithm under three threshold animal models. The first model included effects of stud, mare's birth period and interaction between them as fixed. The second model included the interaction between the stud and mare's birth period as random, whereas in the third model stud effects were also treated as random. The models were compared based on the deviances. Phenotypic and genetic trends were evaluated as well. The twinning rate was 3.5%. The study involved 443 twins and two triplets. Heritability estimates varied from 0.24 (model III) to 0.29 (model I), whereas repeatability estimates ranged from 0.33 (model I) to 0.34 (model III). The fluctuations of average annual genetic effects may result from segregation of the major gene in the population. PMID- 16706922 TI - Genetic and environmental correlations between longevity and litter size in rabbits. AB - An estimation of the genetic and environmental correlations between prolificacy (kits born alive and number of kits at weaning) and functional longevity in a population selected for litter size at weaning since 1992 was carried out. The method is only an approximation, but suggests that genetically longevity and litter size are not antagonistic objectives in breeding programmes, because the estimated genetic correlations are not significantly different from zero. PMID- 16706924 TI - Transmission disequilibrium test for quantitative trait loci detection in livestock populations. AB - The performance of several transmission disequilibrium tests (TDT) for detection of quantitative trait loci (QTL) in data structures typical of outbred livestock populations were investigated. Factorial mating designs were simulated with 10 sires mated to either 50 or 200 dams, each family having five or eight full sibs. A single marker and QTL, both bi-allelic, were simulated using a disequilibrium coefficient based on complete initial disequilibrium and 50 generations of recombination [i.e. D = D(0)(1 - theta)50], where theta is the recombination fraction between marker and QTL. The QTL explained either 10% (small QTL) or 30% (large QTL) of the genetic variance for a trait with heritability of 0.3. Methods were: TDT for QTL (Q-TDT; both parents known), 1-TDT (only one parent known) and sibling-based TDT (S-TDT; neither parent known, but sibs available). All were found to be effective tests for association and linkage between the QTL and a tightly linked marker (theta < 0.02) in these designs. For a large QTL, theta = 0.01, and five full sibs per family, the empirical power for Q-TDT, 1-TDT and S TDT was 0.966, 0.602 and 0.974, respectively, in a large population, versus 0.700, 0.414 and 0.654, respectively, in a small population. For a small QTL effect, theta = 0.01, large population the empirical power of these tests were 0.709, 0.287 and 0.634. The power of Q-TDT, 1-TDT and S-TDT was satisfactory for large populations, for QTL with large effects and for five full sibs per family. The 1-TDT based on a linear model was more powerful than the normal 1-TDT. The empirical power for Q-TDT and 1-TDT with a linear model was 0.978 and 0.995 respectively. TDT based on analogous linear models, incorporating the polygenic covariance structure, provided only small increases in power compared with the usual TDT for QTL. PMID- 16706925 TI - Investigation of two candidate genes for meat quality traits in a quantitative trait locus region on SSC6: the porcine short heterodimer partner and heart fatty acid binding protein genes. AB - A highly significant quantitative trait locus (QTL) on pig chromosome 6, affecting intramuscular fat (IMF), has previously been detected by our group and others. Two genes of positional and biological interest, the small heterodimer partner (SHP; NR0B2) and the heart fatty acid binding protein (FABP3; H-FABP), were investigated for meat quality traits and IMF respectively. SHP was partially sequenced (GenBank: DQ002896 and DQ002897) and mapped to the QTL region on porcine chromosome 6, affecting IMF. The map shows no recombination between SHP and FABP3, which was previously mapped to the same QTL region. Twelve single nucleotide polymorphisms were detected in the sequenced region of SHP gene. Haplotype information was used to investigate association between genetic variation and different meat quality traits. SHP haplotype combinations were found to have significant effect on connective tissue. However, further studies are needed to evaluate this possible association more effectively. The FABP3 is involved in fatty acid transport and has been studied as a candidate gene for IMF by several research groups. In our study, FABP3 genotypes were confirmed to be significantly associated with IMF in pigs. The average content of IMF in our population was 1.6%, which may indicate that the FABP3 polymorphism explains as much as 30-35% of the variation in IMF in our pig cross-population. PMID- 16706926 TI - New SNPs in the IGF2 gene and association between this gene and backfat thickness and lean meat content in Large White pigs. AB - IGF2-in3-G3072A is a causative mutation for paternally expressed quantitative trait loci on the p arm of porcine chromosome 2 with substantial effect on muscle growth and backfat thickness. The linkage disequilibrium between IGF2-in3-G3072A and IGF2-in7-G162C (IGF2-NciI) in four breeds and associations between these polymorphisms and growth and meat performance in pigs of the Large White breed were analysed. A significant effect of these polymorphisms on backfat thickness and lean meat content was found. In addition, we identified two new single nucleotide polymorphisms (SNPs) in intron 7 of the gene. The existence of complete linkage disequilibrium between IGF2-in3-G3072A locus in the population under study where the locus segregated and SNPs in intron 7 of the IGF2 gene detectable with simple and reliable polymerase chain reaction-restriction fragment length polymorphism techniques (G162C, C179G and G186T) offer possibilities to use these SNPs for genotyping of quantitative trait nucleotide in Large White and Landrace breeds. PMID- 16706927 TI - Identification and characterization of polymorphisms within the 5' flanking region, first exon and part of first intron of bovine GH gene. AB - The aim of the present study was to identify and characterize polymorphisms within the 5' flanking region, first exon and part of first intron of the bovine growth hormone gene among different beef cattle breeds: Nelore (n = 25), Simmental (n = 39), Simbrasil (n = 24), Simmental x Nelore (n = 30), Canchim x Nelore (n = 30) and Angus x Nelore (n = 30). Two DNA fragments (GH1, 464 bp and GH2, 453 bp) were amplified by polymerase chain reaction and then used for polymorphism identification by SSCP. Within the GH1 fragment, five polymorphisms were identified, corresponding to three different alleles: GH1.1, GH1.2 and GH1.3 (GenBank: AY662648, AY662649 and AY662650, respectively). These allele sequences were aligned and compared with bovine GH gene nucleotide sequence (GenBank: M57764 and AF118837), resulting in the identification of five insertion/deletions (INDELs) and five single nucleotide polymorphisms (SNPs). In the GH2 fragment two alleles were identified, GH2.1 and GH2.2 (GenBank: AY662651 and AY662652, respectively). The allele sequences were compared with GenBank sequences (M57764, AF007750 and AH009106) and three INDELs and four SNPs were identified. In conclusion, we were able to identify six new polymorphisms of the bovine GH gene (one INDEL and five SNPs), which can be used as molecular markers in genetic studies. PMID- 16706928 TI - Isolation and characterization of the canine NKX2-5 gene. AB - We report the first isolation and characterization of the canine NKX2-5 gene. This canine homologue has high homology in genomic structure and functional domains to other NKX2-5 across a number of different species. Given the critical role of NKX2-5 in cardiac morphogenesis as seen in human and mouse studies of congenital heart defects, the availability of the canine NKX2-5 provides a good starting point for identifying mutations that may be responsible for certain forms of canine congenital heart defects. PMID- 16706929 TI - An approach to the management of leukemia in the developing world. AB - Management of leukemia is expensive. In the developing world where there are significant resource constraints, how does the physician manage leukemia? Government health budgets need to put their money into areas where a small investment is likely to benefit a large proportion of its people and these would include immunization, sanitation, family planning and treatment of communicable diseases and the cost of expensive treatments would have to be borne by the patient and family. However, in many of the emerging economies in the developing world, society is three-tiered in terms of resources with a small proportion being able to afford the best treatment, the middle class with the ability to meet a significant proportion of the healthcare costs on their own and the poor who depend wholly on what the Government health budget can afford. Providing an accurate diagnosis is possible even with limited resources and this must be made available to all. Sometimes a precise diagnosis even at the molecular level is more important in the developing world so that resources are not wasted on inappropriate treatments. Choosing a protocol for management based on resources available for the individual patient would appear ethically unacceptable, but is probably the only solution when wealth is unevenly distributed in society. This paper illustrates an approach to the management of leukemia in the developing world that has evolved with experience over two decades in a tertiary referral hospital in India. PMID- 16706931 TI - Translocation between chromosome 5q35 and chromosome 11q13-- an unusual cytogenetic finding in a primary refractory acute myeloid leukemia. AB - Cytogenetic abnormalities are observed in approximately two-thirds of patients with acute myeloid leukemia (AML). Chromosome rearrangements are associated with specific subtypes of AML and associated prognosis. We report a patient with AML, M2, who was primarily refractory to standard induction chemotherapy with idarubicin and cytarabine. Flow cytometry of a bone marrow aspirate showed aberrant expression of B-cell markers including CD19. Cytogenetic studies disclosed a translocation between 5q35 and 11q13. Fluorescence in situ hybridization analyses demonstrated that neither the NSD1 nor MLL genes were involved in this case. Further study is required to define conclusively the genes involved and their contribution to pathogenesis in this case. PMID- 16706930 TI - Methylenetetrahydrofolate reductase A1298C genotypes are associated with the risks of acute lymphoblastic leukaemia and chronic myelogenous leukaemia in the Korean population. AB - Methylenetetrahydrofolate reductase (MTHFR) is a key enzyme involved in folate metabolism, DNA methylation and synthesis. We investigated the association between MTHFR polymorphisms and the risks of acute and chronic leukaemias. MTHFR C677T and A1298C were genotyped in 396 Korean individuals using multiplex polymerase chain reaction/restriction fragment-length polymorphism. They were acute lymphoblastic leukaemia (ALL, n = 89), acute myeloid leukaemia (AML, n = 55), biphenotypic acute leukaemia (n = 12), chronic myelogenous leukaemia (CML, n = 40), and normal controls (n = 200). C677T genotypes were not associated with the risk of each disease. A1298C variants, however, significantly decreased the risks of ALL and CML compared with 1298AA. Odds ratios and 95% confidence intervals of 1298AC and 1298AC + CC were 0.53 (0.31-0.93) and 0.54 (0.31-0.93) in ALL, and 0.34 (0.14-0.80) and 0.40 (0.18-0.89) in CML, respectively, compared with 1298AA. These findings demonstrate that the development of ALL and CML is more dependent on folate status, and more susceptible to DNA instability than that of AML. In addition, A1298C rather than C677T may be a more important genetic risk modifier in leukaemogenesis at least in the Korean population. PMID- 16706932 TI - Investigation of monoclonal gammopathy of undetermined significance: a single centre study. AB - Summary The main characteristic of monoclonal gammopathies (MG) is the presence of an increased amount of both electrophoretically and immunologically homogeneous immunoglobulins (M component). According to the WHO classification, monoclonal gammopathies are classified among the non-Hodgkin's lymphomas as 'plasma cell dyscrasias'. The unknown behaviour state, so-called MGUS (monoclonal gammopathy of undetermined significance), is distinguished from the malignant diseases. We investigated the frequency and features of MG and MGUS by reviewing the serum immunochemistry protein analyses between 1998 and 2004. Among 18,642 analyses, MG was found in 1,983 cases (10.39%) derived from one or more samples of 416 patients. Case histories of 340 patients were analysed. A malignant lymphoproliferative disease was proved in 171 cases, while in 169 cases the behaviour of the gammopathy was unknown. In 65 cases, the disease was possibly not related to MG. Mean follow-up time of the 65 patients with MGUS was 42 (9-81) months. During the follow-up period seven patients progressed into a malignant lymphoproliferative disorder-- mean probability of the malignant transformation was 3.07%/year and it occurred more frequently in the presence of immunoglobulin A isotype. There was no correlation between the progression of the disease and other laboratory findings. Besides the analyses of MG-related diseases, we focus on the malignant transformation of MGUS and on the importance of regular clinical and laboratory control examinations. PMID- 16706933 TI - Hematological malignancy and pregnancy: a single-institution experience of 21 cases. AB - The incidence of hematological malignancies during pregnancy is low, and treatment in this setting is problematic. This study observed 21 pregnancies in 18 patients with hematological malignancies. Patients' ages were between 19 and 43 (median 25) years. Two pregnancies ended with spontaneous abortion, one pregnancy ended with in utero death, three therapeutic abortions were carried out, and 15 infants were born alive but three of them died later. The median birth weight was 2.47 kg. Twelve babies survived to a median age of 36 (range 4 117) months. Eight babies were exposed to chemotherapy during the in utero period. One baby was exposed to chemotherapy during all the trimesters and was born prematurely and later died because of intracranial bleeding. Four babies were exposed to chemotherapy during the first trimester, one of them had low birth weight and floating thumb malformation, two of them had only low birth weight, and one was born healthy, but died at 3 months of age as a result of severe gastroenteritis. Two babies were exposed to chemotherapy during the second and third trimesters; one of them had low birth weight, and the other pregnancy ended in in utero death. One infant was exposed to chemotherapy during the third trimester and was born at term, but died because of pulmonary hemorrhage. We concluded that chemotherapy during all trimesters of pregnancy carries a significant risk for an unfavorable outcome. PMID- 16706934 TI - Effect of ICAM-1 and LFA-1 in hyperleukocytic acute myeloid leukaemia. AB - Acute hyperleukocytic leukemia [AHL; WBC count >100 x 10(9)/l] is associated with a life-threatening complication. The mechanisms of hyperleukocytosis in acute myeloid leukaemia (AML) remain unclear. However, the interaction of intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function-associated antigen-1 (LFA-1) plays an important role in the adhesion and migration of normal leukocytes and AML cells. Therefore, effects of ICAM-1 and LFA-1 were studied in hyperleukocytic AML. The adhesion of hyperleukocytic AML blasts and human umbilical vein endothelial cells (HUVECs) was significantly increased compared with that of blasts from non-hyperleukocytic AML (WBC < 100 x 10(9)/l). The adhesion of normal neutrophils and HUVECs treated with hyperleukocytic AML blast supernatant was increased significantly. Finally, we determined the ICAM-1 on the surface of HUVECs treated with the supernatant of hyperleukocytic AML blasts and LFA-1 on hyperleukocytic AML blasts by flow cytometry. It showed that the ICAM-1 expression on the surface of the HUVECs treated with hyperleukocytic AML blast supernatant for 24 h could be increased, and the expression of LFA-1 on hyperleukocytic AML was also increased significantly. Our data show that hyperleukocytic AML blasts stimulate the endothelium to secrete more ICAM-1 and promote their own adhesion to vascular endothelium, suggesting that ICAM-1 and LFA-1 may have a role in hyperleukocytic AML. PMID- 16706935 TI - Haematological parameters in elite rugby players during a competitive season. AB - We assessed haematological parameters and possible modifications in elite rugby players throughout a competitive season for increasing the knowledge of physiological characteristics of these sportsmen. Blood samples were collected from the members of the Italian National rugby team at four consecutive training camps during a whole competitive season. Forty-four athletes were recruited for the first camp, 36 for the second, 30 for the third and 32 for the fourth. Due to turnover of the subjects during the season only 13 athletes could be examined at all four camps, and another six in the first three camps. Therefore, we selected the data of these 19 athletes. Iron and transferrin saturation were stable, whilst ferritin increased at the end of the season. The modifications of the soluble transferrin receptor (sTFR) were linked to those of haematocrit: sTFR increased after training and during the competition period when haemoglobin and haematocrit decreased, and decreased at the end of the season. Haemoglobin and haematocrit showed slightly higher levels during the first part of the season and decreased in the second half, when physical demand was high, as demonstrated by biochemical additional tests. Leucocytes and platelets were stable throughout the season. Haematological and iron metabolism parameters in the elite rugby players examined during a whole season fall within physiological range of values. The variability of the parameters during the season is related to training and competition workload. Reticulocytes and sTFR are the most sensitive parameters for studying the iron metabolism of the athletes. PMID- 16706936 TI - Myeloablative allogeneic stem cell transplantation for advanced stage multiple myeloma: very long-term follow up of a single center experience. AB - We aimed to review the long-term outcome of myeloablative allogeneic stem cell transplantation (SCT) performed for multiple myeloma (MM) at our institution. Records of all patients who received standard myeloablative allogeneic SCT for MM were retrospectively reviewed. Overall survival (OS), progression-free survival (PFS), and event-free survival (EFS) were calculated using the Kaplan-Meier method. In total 37 transplants had been performed. Median follow up post-SCT was 108 months (range: 33-148). The majority of patients suffered advanced stage disease and/or had received multiple prior therapies prior to SCT. Transplant related mortality (TRM) at 100 days was 32%. Grades 2-4 acute graft-vs.-host disease (GVHD) occurred in 18 patients (49%), and extensive stage chronic GVHD in seven (28%) of 25 patients surviving greater than day 100. Median OS, PFS, and EFS were 28 months, 66 months and 13 months, respectively, with 5 year OS, PFS, and EFS 40%, 54% and 24%. Our results suggest that allogeneic SCT, even when performed in advanced stage, heavily pretreated MM, still results in long-term EFS in a significant minority of patients. Efforts should continue on alternative allogeneic SCT approaches to reduce the high early TRM rate associated with myeloablative conditioning. PMID- 16706938 TI - Automated separation of cord blood units in top and bottom bags using the Compomat G4. AB - Cord blood (CB) has become a real alternative source of haematopoietic stem cells for bone marrow reconstitution in a variety of malignant disorders. As a response to this increasing activity, CB banks have been developed to guarantee the quality of processed CB units. Volume reduction of CB units maximizes storage space and also has other advantages. The aim of this study was to develop a program for the volume reduction of CB in the Compomat G4 device. We also compared two different top and bottom systems for CB fractionation (Compomat G4 and Optipress II). We empirically designed three different programs for volume reduction of CB with Compomat G4: two for final BC volume of 41 ml (CB1 and CB2) and the other one for buffy coat (BC) volume of 25 ml (CB3). Significantly worse recoveries were achieved for CB processed with program CB3. A RBC depletion of >or=50%, >or=60% and >or=70% were achieved for 67%, 39% and 9% of all units respectively. When comparing Compomat G4 and Optipress II, total nucleated cell recovery was similar for both methods, while lymphocytes recovery was significantly better for Optipress II. PMID- 16706937 TI - Bernard-Soulier syndrome in pregnancy. AB - Bernard-Soulier syndrome (BSS) is a rare autosomal recessively inherited bleeding disorder. Pregnancy in patients with BSS is characterized by ante-, intra-, or postpartum haemorrhage, which may be delayed and severe. There is no consensus in the management of BSS in pregnancy and so far only 16 pregnancies in nine patients have been described. We report a further three pregnancies in two women with the syndrome. We also outline our management of pregnant patients with BSS. PMID- 16706939 TI - Imatinib-associated neutropenia may not be overcome by filgrastim treatment in patients with blastic phase of chronic myeloid leukaemia. AB - Imatinib mesylate is a very effective treatment in patients with Philadelphia (Ph)-positive chronic myeloid leukaemia (CML). However, in patients with advanced phase CML, it is still unclear whether, in the presence of myelosuppression, therapy with imatinib should be continued. It has been reported that intermittent filgrastim treatment may overcome imatinib-associated neutropenia and allow improved delivery of imatinib. Such combined sequential treatment is theoretically attractive as it may lead to better disease response. Here, we report a patient with blastic phase CML who developed severe and prolonged myelosuppression during imatinib treatment. Despite cessation of imatinib and 2 months of filgrastim therapy neither recurrence of Ph-positive or Ph-negative cells occurred. We conclude that filgrastim treatment may not always reverse imatinib-associated neutropenia therefore the decision of continued imatinib therapy in patients with advanced CML should be taken with caution. PMID- 16706940 TI - Accelerated hepatitis C virus replication with rituximab treatment in a non Hodgkin's lymphoma patient. AB - The treatment of patients with non-Hodgkin's lymphoma (NHL) may be complicated by concomitant chronic hepatitis C virus (HCV) infection. Recent data suggest that HCV may also be a contributing factor to the development of this disease. Although antiviral treatment has occasionally been reported to result in the regression of lymphoma in patients with HCV infection, the importance of the control of this infection on the prognosis of lymphoma needs to be defined. Here we report a patient with diffuse large B-cell lymphoma who presented with a mass in her left breast. She had had HCV-related liver cirrhosis for 6 years. She was given rituximab monotherapy for three consecutive weeks, but treatment had to be discontinued as a result of hematological toxicity. HCV viral load also increased, but then decreased gradually after rituximab was stopped. She could be given no further therapy. Six months later she presented with spinal involvement with infiltration of the cauda equina. Though cranial-spinal radiotherapy and steroids were started, she died shortly thereafter. Though rituximab is an invaluable drug in the treatment of B-cell lymphomas, we believe that the use of such agents with potentially long-lasting effects on B lymphocytes requires extended vigilance for accelerated replication of hepatitis B and C viruses. PMID- 16706941 TI - Extramedullary blast crises in CML patients in complete hematological remission treated with imatinib mesylate. PMID- 16706942 TI - Incidence of cancer among female flight attendants: a meta-analysis. AB - BACKGROUND: Airline flight personnel work in a unique environment with exposure to known or suspected carcinogens and mutagens including ionizing cosmic radiation. A meta-analysis was conducted to study whether the occupational exposure of female flight attendants (FA) increased their relative risk of cancer incidence. METHODS: A bibliographical computer search from 1966 to 2005 of cancer incidence cohort studies of female FA was performed. Combined relative risks (RRc) in cancer incidence were calculated by means of meta-analysis. RESULTS: RRc and 95% confidence interval (CI) for malignant melanoma and breast cancer in female FA were 2.13 (95% CI: 1.58-2.88) and 1.41 (1.22-1.62) (p < 0.0001). Excess risk was not significant for all-site cancer with RRc of 1.10 (0.99-1.21). CONCLUSIONS: The meta-analysis confirmed the significantly increased risks for malignant melanoma and breast cancer in female FA. Increased exposure to cosmic radiation during flight has been suggested as a potential occupational risk factor. Ultraviolet radiation exposure on board seems an unlikely occupational risk, but nonoccupational leisure time sun exposure is a possible risk factor. The etiology of the observed increase in incidence of some cancers remains controversial because assessment of possible confounders, especially nonoccupational exposure factors, has thus far been limited. PMID- 16706943 TI - Delivery of medical care for migrants in Germany: delay of diagnosis and treatment. AB - BACKGROUND: Migrants form 9% of Germany's population and 13% of its capital Berlin. Only limited data are available regarding general health status and prevalence of tropical diseases among migrants in Germany. This study was conducted to investigate the spectrum and frequency of tropical diseases among migrants in Berlin and to evaluate the quality of the medical care provided. The necessity of a routine screening for tropical diseases among migrants was assessed. METHODS: Anonymized data of migrants presenting to the Berlin Institute of Tropical Medicine between 1999 and 2004 with a stay in Germany below 1 year (n= 153) were analyzed. RESULTS: Of all examined migrants, 48% needed immediate medical treatment and 38% carried an infectious disease, mainly nematodes and intestinal protozoa. 19% suffered from a noninfectious disease, mainly anemia, and 12% were transferred to other specialists for further investigation. These figures were similar among asymptomatic and symptomatic patients. The median duration of stay in Germany until presentation was 42 days. While 40% of the migrants were examined within the first 4 weeks of their stay, 20% had not received a medical examination after 6 months. Of this population, 50% required treatment upon presentation. CONCLUSIONS: The high proportion of delayed diagnosis and treatment indicates a lack of medical service for migrants. While this clearly translates into increased health risks for the individual patient, it also indicates a potential risk for transmission of communicable diseases in the community. The lack of a correlation between symptoms and detected infectious disease indicates the need for a standardized routine screening examination in all migrants. PMID- 16706944 TI - Prospective analysis of parasitic infections in Canadian travelers and immigrants. AB - BACKGROUND: International travel is associated with increased risk of vector borne illnesses, particularly malaria. The objective of this study was to prospectively assess the relative frequency of parasitic diseases in Canadian travelers and to characterize demographic and travel-related predictors of these infections. METHODS: Data on Canadians and new immigrants who crossed international borders and were seen in the Tropical Disease Unit of Toronto General Hospital between November 1997 and June 2003 were prospectively collected and entered into the GeoSentinel Surveillance Network database. RESULTS: Of 3,528 returned Canadian travelers and new immigrants in the database, 1,010 had a parasitic infection diagnosed. Mean age of the 3,528 travelers was 37.3 years, and 42.6% were male. Those diagnosed with parasitic infections were more likely than the remaining cohort to have been traveling for the purpose of immigration (21.1% vs 7.1%, p < 0.001), or visiting friends and relatives (VFR) (17.9% vs 11.8%, p < 0.01). Common parasitic infections included nonhistolytica amebiasis (N= 209), malaria (N= 143), cutaneous larva migrans (N= 105), giardiasis (N= 74), and schistosomiasis (N= 48). CONCLUSIONS: Parasitic infections occurred in 29% of Canadian travelers. New immigrants and VFRs are at increased risk for malaria, as well as protozoal and helminthic infections. PMID- 16706945 TI - Illness in returned travelers and immigrants/refugees: the 6-year experience of two Australian infectious diseases units. AB - BACKGROUND: Data comparing returned travelers and immigrants/refugees managed in a hospital setting is lacking. METHODS: We prospectively collected data on 1,106 patients with an illness likely acquired overseas who presented to two hospital based Australian infectious diseases units over a 6-year period. RESULTS: Eighty three percent of patients were travelers and 17% immigrants/refugees. In travelers, malaria (19%), gastroenteritis/diarrhea (15%), and upper respiratory tract infection (URTI) (7%) were the most common diagnoses. When compared with immigrants/refugees, travelers were significantly more likely to be diagnosed with gastroenteritis/diarrhea [odds ratio (OR) 8], malaria (OR 7), pneumonia (OR 6), URTI (OR 3), skin infection, dengue fever, typhoid/paratyphoid fever, influenza, and rickettsial disease. They were significantly less likely to be diagnosed with leprosy (OR 0.03), chronic hepatitis (OR 0.04), tuberculosis (OR 0.05), schistosomiasis (OR 0.3), and helminthic infection (OR 0.3). In addition, travelers were more likely to present within 1 month of entry into Australia (OR 96), and have fever (OR 8), skin (OR 6), gastrointestinal (OR 5), or neurological symptoms (OR 5) but were less likely to be asymptomatic (OR 0.1) or have anaemia (OR 0.4) or eosinophilia (OR 0.3). Diseases in travelers were more likely to have been acquired via a vector (OR 13) or food and water (OR 4), and less likely to have been acquired via the respiratory (OR 0.2) or skin (OR 0.6) routes. We also found that travel destination and classification of traveler can significantly influence the likelihood of a specific diagnosis in travelers. Six percent of travelers developed a potentially vaccine-preventable disease, with failure to vaccinate occurring in 31% of these cases in the pretravel medical consultation. CONCLUSIONS: There are important differences in the spectrum of illness, clinical features, and mode of disease transmission between returned travelers and immigrants/refugees presenting to hospital-based Australian infectious diseases units with an illness acquired overseas. PMID- 16706946 TI - Imported malaria in Jakarta, Indonesia: passive surveillance of returned travelers and military members postdeployment. AB - BACKGROUND: Autochthonous malaria does not currently occur in Jakarta, the most populous city in Indonesia. Military, forestry, mining, and tourist activities draw Jakarta residents to distant parts of the archipelago with high rates of malaria. Although malaria is a reportable disease in Jakarta, little has been published. METHODS: We collected demographic and travel information from patients in Jakarta with microscopically confirmed malaria from January 2004 to February 2005, using a standardized data collection form. These results were compared to regional rainfall statistics and transit patterns of Jakarta residents to and from rural areas. RESULTS: Data from 240 patients were collected. Aceh Province was the travel destination most commonly recorded for military members, while Papua and Bangka Island were the most frequently cited by civilians. Plasmodium falciparum accounted for 53% of cases, of which 15% had detectable gametocytemia. The most common admission diagnoses were malaria (39%), febrile illness not otherwise specified (23%), viral hepatitis (19%), and dengue (11%). The median time from admission to microscopic diagnosis was 2 days for civilian patients and 2.5 days for military patients. The highest number of cases occurred in May, July, and December with the nadir in October. CONCLUSIONS: The diagnosis of malaria may be overlooked and therefore delayed, in nonendemic areas such as Jakarta. Travel destinations associated with contracting malaria vary significantly for civilian and military populations. The factors affecting the peak months of importation likely include rainfall, holiday transit, military flight availability, and referral center locations. PMID- 16706947 TI - Malaria chemoprophylaxis advice: survey of South African community pharmacists' knowledge and practices. AB - BACKGROUND: Over 3 million South African residents travel to malarious areas annually. Given pharmacists' ready accessibility and travel clinics' incapacity to service total potential demand, South African community pharmacists' malaria chemoprophylaxis knowledge and practice were assessed. METHODS: Covert survey. Pharmacies were approached at random and asked for malaria chemoprophylaxis recommendations. A standard questionnaire indicating a 3-day stay in either Maputo (N= 43; malarious) or Harare (N= 25; non-malarious) was used. RESULTS: Maputo group: 41/43 (95%) pharmacists correctly identified need for chemoprophylaxis; 3/41 (7%) recommended an ineffective drug. Eight of 41 (20%) enquired about diving, and 6/41 (15%) enquired about epilepsy or mental illness; despite positive responses mefloquine, was nevertheless recommended. Harare group: 12/25 (48%) incorrectly advised chemoprophylaxis was necessary; 4/12 (33%) sought contraindications; all prophylaxis recommended was considered effective. Overall, 54/68 (79%) pharmacists correctly determined whether chemoprophylaxis was required or not; 6/53 (11%) of all recommended chemoprophylaxis included "alternative" antimalarials; 1/68 (1%) consulted external advice before making recommendations; 11/53 (21%) referred travelers to a physician; 1/68 (1%) referred to a travel clinic. Pharmacies were significantly more likely (Fisher exact, p < 0.0001) to recommend unnecessary prophylaxis (12/25; 48%) than to advise against necessary prophylaxis (2/43; 5%). CONCLUSIONS: Pharmacists are willing to give malaria chemoprophylaxis advice but appear to overprescribe; although unnecessary adverse events may result, this may be the preferred error in a falciparum-dominated region. Pharmacists' knowledge of contraindications, willingness to consult external resources, and knowledge of antimalarials' effectiveness could be improved. Pharmacists appear unwilling to refer to travel clinics. An effective intervention to improve the safety and accuracy of pharmacists' advice might be the provision of a simple aid listing effective antimalarials and their contraindications, illustrated with a malaria risk map. Improving the safety and accuracy of pharmacists' advice would increase significantly travelers' access to reliable travel health information. PMID- 16706948 TI - A randomized, controlled field trial for the prevention of jellyfish stings with a topical sting inhibitor. AB - BACKGROUND: Jellyfish stings are a common occurrence among ocean goers worldwide with an estimated 150 million envenomations annually. Fatalities and hospitalizations occur annually, particularly in the Indo-Pacific regions. A new topical jellyfish sting inhibitor based on the mucous coating of the clown fish prevents 85% of jellyfish stings in laboratory settings. The field effectiveness is unknown. The objective is to evaluate the field efficacy of the jellyfish sting inhibitor, Safe Sea. METHODS: A double-blind, randomized, placebo controlled trial occurred at the Dry Tortugas National Park, FL, USA and Sapodilla Cayes, Belize. Participants were healthy volunteers planning to snorkel for 30 to 45 minutes. Ten minutes prior to swimming, each participant was directly observed applying a blinded sample of Safe Sea (Nidaria Technology Ltd, Jordan Valley, Israel) to one side of their body and a blinded sample of Coppertone (Schering-Plough, Kenilworth, NJ, USA) to the contralateral side as placebo control. Masked 26 g samples of both Safe Sea SPF15 and Coppertone SPF15 were provided in identical containers to achieve 2 mg/cm(2) coverage. Sides were randomly chosen by participants. The incidence of jellyfish stings was the main outcome measure. This was assessed by participant interview and examination as subjects exited the water. RESULTS: A total of 82 observed water exposures occurred. Thirteen jellyfish stings occurred during the study period for a 16% incidence. Eleven jellyfish stings occurred with placebo, two with the sting inhibitor, resulting in a relative risk reduction of 82% (95% confidence interval: 21%-96%; p= 0.02). No seabather's eruption or side effects occurred. CONCLUSIONS: Safe Sea is a topical barrier cream effective at preventing >80% jellyfish stings under real-world conditions. PMID- 16706949 TI - Mefloquine-induced pneumonitis. AB - Mefloquine is indicated as oral treatment and as prophylaxis for malaria in areas where chloroquine-resistant malaria is present. Gastrointestinal and neuropsychiatric side effects of mefloquine are well known. More severe neuropsychiatric disorders such as psychosis, depression, hallucinations, and seizures are also reported in the literature. We are reporting a case of drug induced pneumonia due to mefloquine. This diagnosis was confirmed 4 months after the adverse event, after restarting the same malaria prophylaxis, which could be considered as an unintentional provocation test. This is the third case report in the literature of acute lung injury caused by mefloquine. PMID- 16706950 TI - Tropical sprue in two foreign residents, with evidence of Tropheryma whippelii in one case. AB - Tropical sprue is a rare disease in travelers. Its etiology remains unclear. We report two cases of tropical sprue occurring in long-term residents in Nepal and Cameroon. In one case, Tropheryma whippelii, the agent of Whipple's disease, was identified. Many infectious agents have been suggested to be the etiological agent of tropical sprue, but no association with Whipple's disease has yet been reported. PMID- 16706951 TI - Acalculous cholecystitis in a patient with Plasmodium falciparum infection: a case report and literature review. AB - Acute acalculous cholecystitis (AAC) can occur without gallstones in critically ill or injured patients and has also been associated with various infectious agents.(1-4) We report here a case of AAC in a patient with Plasmodium falciparum malaria. PMID- 16706952 TI - Disseminated Penicillium marneffei infection in an HIV-positive Italian patient and a review of cases reported outside endemic regions. AB - We describe a case of disseminated Penicillium marneffei in a human immunodeficiency virus (HIV)-positive Italian man who stayed for 4 years in Chiang Ray province, northern Thailand. A review of the literature shows that penicilliosis, although unusual, may represent an emerging opportunistic infection among HIV-positive people traveling to endemic areas. PMID- 16706954 TI - A tale of two societies. PMID- 16706955 TI - Remembrances of factor VIII. Part 2: the path to mutation discovery. PMID- 16706956 TI - Pharmacoeconomics of anticoagulation therapy for stroke prevention in atrial fibrillation: a review. AB - INTRODUCTION: Atrial fibrillation (AF) increases the risk of ischemic stroke 5 fold and may not only be responsible for as many as 15% of all strokes that occur but also for larger and more disabling strokes than those attributable to other causes which increase the associated costs of care. Anticoagulation with warfarin in the target INR of 2.5 is a major clinical challenge in real-life practice, given that the complex relationship between warfarin dosage and response is readily altered by a variety of factors such as concurrent medications, illnesses, genetic influences, and dietary/lifestyle changes. Consequently, INR values are out of the target range approximately half of the time in real-life studies compared to clinical trial setting. Current anticoagulation therapies are less likely to be cost-effective in routine clinical practice and need improvement. The aim of this review is to discuss the pharmacoeconomic consequences of this management strategy by analysing the optimal treatment option within specific age and risk groups, confirming current guidelines for a health economic perspective and considering the economic impact on health care policy. METHODS: An electronic search of the Medline/PubMed database from 1966 to 2005 was performed to identify articles dealing with all pharmacoeconomic aspects of stroke prevention in atrial fibrillation. The following search terms were used: 'atrial fibrillation', 'stroke', 'cost', 'warfarin'. RESULTS: Treatment with warfarin is cost-effective (versus aspirin or no therapy) in patients with AF at moderate-to-high risk of stroke. The cost-effectiveness of anticoagulation therapy is driven by the achieved risk reduction rather than the potential benefits estimated from clinical trials. Failure to maintain optimal anticoagulation places patients at risk of complications, the management of which is a significant cost driver. CONCLUSION: Improvement could be achieved by optimising physicians and patient's knowledge driven through prevention campaigns by health care policy. PMID- 16706957 TI - von Willebrand factor, endothelial dysfunction, and cardiovascular disease. AB - von Willebrand factor (VWF), a glycoprotein involved in arterial thrombus formation, is released into the circulation by secretion from endothelial cells. Plasma VWF levels are determined by genetic factors including ABO blood groups and VWF mutations, and by non-genetic factors including aging, impaired nitric oxide production, inflammation, free radical production and diabetes. Plasma VWF levels have been proposed as a risk factor for cardiovascular events. Although they are only weakly associated with the risk of coronary heart disease (CHD) in the general population, they are a more promising CHD risk factor in high-risk populations with previous cardiovascular events, diabetes or old age. However, is it still unclear whether VWF levels directly determine the rate and severity of arterial thrombus formation or whether they merely reflect alteration in other endothelial functions. The future status of VWF levels as a cardiovascular risk factor depends on additional studies on the genetic determinants of both VWF levels and cardiovascular outcomes. Further studies on VWF levels as a predictor of the risk of stroke (rather than CHD) in elderly or other high-risk population are also promising. Such studies could lead to the clinical use of plasma VWF levels to refine the estimation of the cardiovascular risk and of the expected benefit of antithrombotic agents. PMID- 16706958 TI - Polymorphisms and platelet genotyping: the shape of things to come. PMID- 16706959 TI - Definition of novel GP6 polymorphisms and major difference in haplotype frequencies between populations by a combination of in-depth exon resequencing and genotyping with tag single nucleotide polymorphisms. AB - BACKGROUND: Common genetic variants of cell surface receptors contribute to differences in functional responses and disease susceptibility. We have previously shown that single nucleotide polymorphisms (SNPs) in platelet glycoprotein VI (GP6) determine the extent of response to agonist. In addition, SNPs in the GP6 gene have been proposed as risk factors for coronary artery disease. METHODS: To completely characterize genetic variation in the GP6 gene we generated a high-resolution SNP map by sequencing the promoter, exons and consensus splice sequences in 94 non-related Caucasoids. In addition, we sequenced DNA encoding the ligand-binding domains of GP6 from non-human primates to determine the level of evolutionary conservation. RESULTS: Eighteen SNPs were identified, six of which encoded amino acid substitutions in the mature form of the protein. The single non-synonymous SNP identified in the exons encoding the ligand-binding domains, encoding for a 103Leu > Val substitution, resulted in reduced ligand binding. Two common protein isoforms were confirmed in Caucasoid with frequencies of 0.82 and 0.15. Variation at the GP6 locus was characterized further by determining SNP frequency in over 2000 individuals from different ethnic backgrounds. CONCLUSIONS: The SNPs were polymorphic in all populations studied although significant differences in allele frequencies were observed. Twelve additional GP6 protein isoforms were identified from the genotyping results and, despite extensive variation in GP6, the sequence of the ligand binding domains is conserved. Sequences from non-human primates confirmed this observation. These data provide valuable information for the optimal selection of genetic variants for use in future association studies. PMID- 16706960 TI - Can anticoagulant treatment be tailored with biomarkers in patients with venous thromboembolism? PMID- 16706961 TI - D-dimer, factor VIII coagulant activity, low-intensity warfarin and the risk of recurrent venous thromboembolism. AB - BACKGROUND: Elevated plasma D-dimer and factor VIII coagulant activity (FVIIIc) may be associated with the risk of recurrent venous thromboembolism (VTE). OBJECTIVES: To evaluate D-dimer and FVIIIc as risk factors for recurrent VTE and assess the efficacy of extended low-intensity warfarin (target International Normalized Ratio 1.5-2.0) in preventing recurrence by biomarker level. PATIENTS AND METHODS: In the Prevention of Recurrent Venous Thromboembolism trial, 508 idiopathic VTE patients treated for > or = 3 months with full-intensity warfarin, and who had stopped warfarin for 7 weeks on average, were randomized to low intensity warfarin or placebo and followed for 2.1 years for recurrent VTE. Prerandomization blood samples were analysed for D-dimer and FVIIIc. RESULTS: One third of participants had elevated baseline D-dimer (> or = 500 ng mL(-1)) and one-fourth, elevated FVIIIc (> or = 150 IU dL(-1)). Adjusting for other risk factors, the hazard ratios (HRs) for recurrent VTE with elevated D-dimer or FVIIIc were 2.0 [95% confidence interval (CI) 1.2-3.4] and 1.5 (95% CI 0.8-2.8), respectively. The association of elevated D-dimer with recurrence was larger among patients with one prior VTE (HR 3.2, 95% CI 1.3-8.0) than in patients with more than one event (HR 1.4, 95% CI 0.7-2.2). For patients with one prior VTE on placebo, the annual recurrence incidence was 10.9% with elevated D-dimer and 2.9% with normal values. Low-intensity warfarin was equally effective in recurrence risk reduction in those with normal or elevated biomarkers. CONCLUSIONS: Among patients with idiopathic VTE, measurement of D-dimer, but not FVIIIc, might be useful for risk stratification. The efficacy of extended low-intensity warfarin therapy did not vary by biomarker level. PMID- 16706962 TI - Toward gene therapy for hemophilia A with novel adenoviral vectors: successes and limitations in canine models. PMID- 16706964 TI - Parachutes and prophylaxis: they both work! PMID- 16706963 TI - Helper-dependent adenoviral gene therapy mediates long-term correction of the clotting defect in the canine hemophilia A model. AB - BACKGROUND: Adenoviral vector-mediated gene therapy might have potential for long term correction of the monogenic disease hemophilia A. OBJECTIVE: In this study, we tested the efficacy of administering a helper-dependent adenoviral vector (HDV) designed for maximal liver-restricted canine factor VIII (cFVIII) expression on three out-bred hemophilia A dogs. METHODS: Three FVIII-deficient animals from the University of North Carolina colony were injected with 1 x 10(12) (Dog A), and 3 x 10(12) (Dog B and C) vp kg(-1) helper-dependent adenoviral vector, and we performed systematic analysis of toxicity, persistence of therapeutic gene expression, and molecular analysis of gene transfer. RESULTS: We observed acute dose-dependent elevation in liver enzymes and thrombocytopenia after injection, although both were transient and resolved within 2 weeks. The whole blood clotting time (WBCT), plasma FVIII concentration, FVIII activity, and activated partial thromboplastin time in all animals improved significantly after treatment, and two animals receiving a higher dose reached near normal WBCT with low-level FVIII activity until terminal sacrifice at 3 months, and 2 years. Importantly, the treated dogs suffered no bleeding events after injection. Moreover, we observed persistent vector-specific DNA and RNA in liver tissue collected from one high-dose animal at days 18 and 79, and could not detect the formation of inhibitory antibodies. CONCLUSION: Although vector-associated toxicity remains an obstacle, a single injection of HDV led to long-term transgene expression and vector persistence in two FVIII-deficient animals with conversion of their severe phenotype to a moderate one. PMID- 16706965 TI - Tailored prophylaxis in severe hemophilia A: interim results from the first 5 years of the Canadian Hemophilia Primary Prophylaxis Study. AB - BACKGROUND: Prophylactic treatment for severe hemophilia A is likely to be more effective than treatment when bleeding occurs, however, prophylaxis is costly. We studied an inception cohort of 25 boys using a tailored prophylaxis approach to see if clotting factor use could be reduced with acceptable outcomes. METHODS: Ten Canadian centers enrolled subjects in this 5-year study. Children were followed every 3 months at a comprehensive care hemophilia clinic. They were initially treated with once-weekly clotting factor; the frequency was escalated in a stepwise fashion if unacceptable bleeding occurred. Bleeding frequency, target joint development, physiotherapy and radiographic outcomes, as well as resource utilization, were determined prospectively. RESULTS: The median follow up time was 4.1 years (total 96.9 person-years). The median time to escalate to twice-weekly therapy was 3.42 years (lower 95% confidence limit 2.05 years). Nine subjects developed target joints at a rate of 0.09 per person-year. There was an average of 1.2 joint bleeds per person-year. The cohort consumed on average 3656 IU kg(-1)year(-1) of factor (F) VIII. Ten subjects required central venous catheters (three while on study); no complications of these devices were seen. One subject developed a transient FVIII inhibitor. End-of-study joint examination scores--both clinically and radiographically--were normal or near-normal. CONCLUSIONS: Most boys with severe hemophilia A will probably have little bleeding and good joint function with tailored prophylaxis, while infusing less FVIII than usually required for traditional prophylaxis. PMID- 16706966 TI - Screening for children from families with Rendu-Osler-Weber disease: from geneticist to clinician. AB - BACKGROUND: Rendu-Osler-Weber syndrome, or hereditary hemorrhagic telangiectasia (HHT), is an autosomal dominant vascular disorder. The syndrome is characterized by telangiectases and arteriovenous malformations (AVMs) affecting skin, mucosae and internal organs. AVMs often remain clinically silent until provoking sudden serious complications, responsible for important morbidity and mortality which can occur both in adulthood and in children. The incidence of AVMs in HHT pediatric populations is unknown. OBJECTIVE: To describe the screening protocol performed in the first genotypically confirmed HHT pediatric population and to estimate the incidence of occult brain, lung and liver AVMs and the different disease phenotypes. MATERIALS AND METHODS: Molecular analysis was performed on 35 children, both symptomatic and asymptomatic, who were family members of probands with a previously identified mutation. Clinical-instrumental examination was performed on the mutation positive cases. Nasal telangiectases were investigated by anterior rhinoscopy. Contrast echocardiography and high resolution thoracic multislice computed tomography (CT) were performed to detect pulmonary arteriovenous malformations (PAVMs), and echo-color Doppler, and abdominal CT to detect hepatic arteriovenous malformations (HAVMs). Brain magnetic resonance imaging was utilized to detect cerebral angiopathic involvement. RESULTS: Molecular analysis demonstrated the mutation-carrier status in 22/35 children. Nineteen children, 12 of whom had epistaxis, positive to molecular testing underwent clinical evaluation. Nasal teleangiectases were found in 68%, mucocutaneous telangiectases (fingers, lips and oral cavity) in 79%, PAVMs in 53%, HAVMs in 47% and cerebral anteriovenous malformations and/or cerebral ischemic changes secondary to PAVMs in 12%. CONCLUSIONS: We evidenced a high incidence of HHT children with occult visceral lesions suggesting that a diagnostic screening may be indicated to appropriately treat brain and lung malformations. PMID- 16706967 TI - Clinical outcomes with unfractionated heparin or low-molecular-weight heparin as bridging therapy in patients on long-term oral anticoagulants: the REGIMEN registry. AB - BACKGROUND: Patients who receive long-term oral anticoagulant (OAC) therapy often require interruption of OAC for an elective surgical or an invasive procedure. Heparin bridging therapy has been used in these situations, although the optimal method has not been established. No large prospective studies have compared unfractionated heparin (UFH) with low-molecular-weight heparin (LMWH) for the perioperative management of patients at risk of thromboembolism requiring temporary interruption of long-term OAC therapy. PATIENTS/METHODS: This multicenter, observational, prospective registry conducted in North America enrolled 901 eligible patients on long-term OAC who required heparin bridging therapy for an elective surgical or invasive procedure. Practice patterns and clinical outcomes were compared between patients who received either UFH alone (n = 180) or LMWH alone (n = 721). RESULTS: Overall, the majority of patients (74.5%) requiring heparin bridging therapy had arterial indications for OAC. LMWH, in mostly twice-daily treatment doses, represented approximately 80% of the study population. LMWH-bridged patients had significantly fewer arterial indications for OAC, a lower mean Charlson comorbidity score, and were less likely to undergo major or cardiothoracic surgery, receive intraprocedural anticoagulants or thrombolytics, or receive general anesthesia than UFH-bridged patients (all P < 0.05). The LMWH group had significantly more bridging therapy completed in an outpatient setting or with a < 24-h hospital stay vs. the UFH group (63.6% vs. 6.1%, P < 0.001). In the LMWH and UFH groups, similar rates of overall adverse events (16.2% vs. 17.1%, respectively, P = 0.81), major composite adverse events (arterial/venous thromboembolism, major bleed, and death; 4.2% vs. 7.9%, respectively, P = 0.07) and major bleeds (3.3% vs. 5.5%, respectively, P = 0.25) were observed. The thromboembolic event rates were 2.4% for UFH and 0.9% for LMWH. Logistic regression analysis revealed that for postoperative heparin use a Charlson comorbidity score > 1 was an independent predictor of a major bleed and that vascular, general, and major surgery were associated with non significant trends towards an increased risk of major bleed. CONCLUSIONS: Treatment-dose LMWH, mostly in the outpatient setting, is used substantially more often than UFH as bridging therapy in patients with predominately arterial indications for OAC. Overall adverse events, including thromboembolism and bleeding, are similar for patients treated with LMWH or UFH. Postoperative heparin bridging should be used with caution in patients with multiple comorbidities and those undergoing vascular, general, and major surgery. These findings need to be confirmed using large randomized trials for specific patient groups undergoing specific procedures. PMID- 16706968 TI - Elevated levels of soluble fibrin or D-dimer indicate high risk of thrombosis. AB - BACKGROUND: Fibrin-related markers such as soluble fibrin (SF) and D-dimer are considered useful for the diagnosis of thrombosis. However, the evidence for diagnosis of thrombosis by fibrin-related markers is not well-established. OBJECTIVE: To evaluate the cutoff values of D-dimer and SF in the diagnosis of thrombosis. PATIENTS AND METHODS: Plasma concentrations of SF and D-dimer were measured in 784 inpatients suspected of having thrombosis between 1 August 2003 and 31 December 2004, and then correlated with thrombosis. RESULTS AND CONCLUSIONS: Plasma concentrations of D-dimer and SF were significantly higher in patients with disseminated intravascular coagulation (DIC), deep vein thrombosis (DVT) and cerebral thrombosis, compared with those in patients without thrombosis. When cutoff values of > 3.0 microg mL(-1) for D-dimer and > 6.0 microg mL(-1) for SF were used for the diagnosis, more than 50% of patients (with the exception of liver transplant patients and postoperative patients) had thrombosis. Receiver operating characteristic analysis showed that SF was more useful than D-dimer for the diagnosis of thrombosis (i.e. DVT and DIC). The cutoff value of D-dimer (7.87 microg mL(-1)) was the same for DVT and DIC, while that of SF was slightly lower for DVT (7.05 microg mL(-1)) than for DIC (8.60 microg mL(-1)). Our findings suggest that high levels of plasma fibrin-related markers reflect high risk for thrombosis. PMID- 16706969 TI - Obesity and risk of venous thromboembolism among postmenopausal women: differential impact of hormone therapy by route of estrogen administration. The ESTHER Study. AB - BACKGROUND: Oral estrogen use and elevated body mass index (BMI) increase the risk of venous thromboembolism (VTE). Recent data suggest that transdermal estrogen might be safe with respect to thrombotic risk. However, the impact of transdermal estrogen on the association between overweight (25 kg m(-2) < BMI < or = 30 kg m(-2)) or obesity (BMI >30 kg m(-2)) and VTE risk has not been investigated. METHODS: We carried a multicenter case-control study of VTE among postmenopausal women aged 45-70 years, between 1999 and 2005, in France. Case population consisted of women with a first documented idiopathic VTE. We recruited 191 hospital cases matched with 416 hospital controls and 62 outpatient cases matched with 181 community controls. RESULTS: The odds ratio (OR) for VTE was 2.5 [95% confidence interval (CI):1.7-3.7] for overweight and 3.9 (95% CI: 2.2-6.9) for obesity. Oral, not transdermal, estrogen was associated with an increased VTE risk (OR = 4.5; 95% CI: 2.6-7.7 and OR = 1.1; 95% CI: 0.7-1.7, respectively). Compared with non-users with normal weight, the combination of oral estrogen use and overweight or obesity further enhanced VTE risk (OR = 10.2; 95% CI: 3.5-30.2 and OR = 20.6; 95% CI: 4.8-88.1, respectively). However, transdermal users with increased BMI had similar risk as non-users with increased BMI (OR = 2.9; 95% CI: 1.5-5.8 and OR = 2.7; 95% CI: 1.7-4.5 respectively for overweight; OR = 5.4; 95% CI: 2.1-14.1 and OR = 4.0; 95% CI: 2.1-7.8 respectively for obesity). CONCLUSIONS: In contrast to oral estrogen, transdermal estrogen does not confer an additional risk of idiopathic VTE in women with increased BMI. The safety of transdermal estrogen on thrombotic risk has to be confirmed. PMID- 16706970 TI - Incorrect use of thromboprophylaxis for venous thromboembolism in medical and surgical patients: results of a multicentric, observational and cross-sectional study in Brazil. AB - OBJECTIVES: Although effective strategies for the prevention of venous thromboembolism (VTE) are widely available, a significant number of patients still develop VTE because appropriate thromboprophylaxis is not correctly prescribed. We conducted this study to estimate the risk profile for VTE and the employment of adequate thromboprophylaxis procedures in patients admitted to hospitals in the state of Sao Paulo, Brazil. METHODS: Four hospitals were included in this study. Data on risk factors for VTE and prescription of pharmacological and non-pharmacological thromboprophylaxis were collected from 1454 randomly chosen patients (589 surgical and 865 clinical). Case report forms were filled according to medical and nursing records. Physicians were unaware of the survey. Three risk assessment models were used: American College of Chest Physicians (ACCP) Guidelines, Caprini score, and the International Union of Angiololy Consensus Statement (IUAS). The ACCP score classifies VTE risk in surgical patients and the others classify VTE risk in surgical and clinical patients. Contingency tables were built presenting the joined distribution of the risk score and the prescription of any pharmacological and non-pharmacological thromboprophylaxis (yes or no). RESULTS: According to the Caprini score, 29% of the patients with the highest risk for VTE were not prescribed any thromboprophylaxis. Considering the patients under moderate, high or highest risk who should be receiving prophylaxis, 37% and 29% were not prescribed thromboprophylaxis according to ACCP (surgical patients) and IUAS risk scores, respectively. In contrast, 27% and 42% of the patients at low risk of VTE, according to Caprini and IUAS scores, respectively, had thromboprophylaxis prescribed. CONCLUSION: Despite the existence of several guidelines, this study demonstrates that adequate thromboprophylaxis is not correctly prescribed: high risk patients are under-treated and low-risk patients are over-treated. This condition must be changed to insure that patients receive adequate treatment for the prevention of thromboembolism. PMID- 16706971 TI - Aspirin resistance detected with aggregometry cannot be explained by cyclooxygenase activity: involvement of other signaling pathway(s) in cardiovascular events of aspirin-treated patients. AB - OBJECTIVES: Although the concept of aspirin resistance is extensively reported in medical literature, its precise mechanisms and clinical outcomes are largely unknown. In this study, we examined individual thromboxane biosynthesis and platelet aggregation in aspirin-treated patients, and whether the results of a platelet aggregation test influenced clinical outcomes. RESULTS: Subjects taking 81 mg of aspirin (n = 50) and controls (n = 38) were evaluated for platelet aggregation and platelet cyclooxygenase-1 (COX-1) activity by measuring collagen induced thromboxane B2 production. For aggregometry, both light transmission (LT) and laser-light scattering methods were employed to quantitatively evaluate aggregate sizes and numbers. Aspirin treatment resulted in the inhibition of collagen-induced platelet aggregation, particularly the transition from small to large platelet aggregates. Although platelet COX-1 activity seemed to be uniformly inhibited in all patients, platelet aggregation studies showed great inter-individual differences; variation in platelet COX-1 activity only accounted for 6-20% of the individual aggregations. Factor analysis revealed the existence of a common factor (other than platelet COX-1) that explained 48.4% of the variations in platelet aggregation induced by collagen, adenosine diphosphate (ADP), and collagen-related peptide. We then prospectively enrolled 136 aspirin treated patients in our study, and we found that being in the upper quartile level of LT, or with large aggregate formation induced by collagen, was an independent risk factor for developing cardiovascular events within 12 months [hazard ratio (HR) = 7.98, P = 0.008 for LT; HR = 7.76, P = 0.007 for large aggregates]. On the other hand, the existence of diabetes mellitus was an independent risk factor for overall outcomes (HR 1.30-11.9, P = 0.015-0.033). CONCLUSIONS: Aspirin resistance expressed as unsuppressed platelet COX-1 activity is a rare condition in an out-patient population. Other factor(s) affecting collagen-induced platelet aggregation may influence early outcomes in aspirin treated patients. PMID- 16706972 TI - Association between patterns of nucleotide variation across the three fibrinogen genes and plasma fibrinogen levels: the Coronary Artery Risk Development in Young Adults (CARDIA) study. AB - BACKGROUND: Previous genotype-phenotype association studies of fibrinogen have been limited by incomplete knowledge of genomic sequence variation within and between major ethnic groups in FGB, FGA, and FGG. METHODS: We characterized the linkage disequilibrium patterns and haplotype structure across the human fibrinogen gene locus in European- and African-American populations. We analyzed the association between common polymorphisms in the fibrinogen genes and circulating levels of both 'functional' fibrinogen (measured by the Clauss clotting rate method) and total fibrinogen (measured by immunonephelometry) in a large, multi-center, bi-racial cohort of young US adults. RESULTS: A common haplotype tagged by the A minor allele of the well-studied FGB-455 G/A promoter polymorphism (FGB 1437) was confirmed to be strongly associated with increased plasma fibrinogen levels. Two non-coding variants specific to African-American chromosomes, FGA 3845 A and FGG 5729 G, were each associated with lower plasma fibrinogen levels. In European-Americans, a common haplotype tagged by FGA Thr312Ala and several other variant alleles across the fibrinogen gene locus was strongly associated with decreased fibrinogen levels as measured by functional assay, but not by immunoassay. Overall, common polymorphisms within the three fibrinogen genes explain < 2% of the variability in plasma fibrinogen concentration. CONCLUSIONS: In young adults, fibrinogen multi-locus genotypes are associated with plasma fibrinogen levels. The specific single nucleotide polymorphism and haplotype patterns for these associations differ according to population and also according to phenotypic assay. It is likely that a substantial proportion of the heritable component of plasma fibrinogen concentration is due to genetic variation outside the three fibrinogen genes. PMID- 16706973 TI - Plasminogen and plasmin activity in patients with coronary artery disease. AB - OBJECTIVE: While coronary artery disease (CAD) is associated with disturbances of the plasma fibrinolytic system, the nature of these disturbances is not fully defined. Fibrinolysis is regulated by plasmin, whose production is mediated by plasminogen activator conversion of plasminogen (Plg) to plasmin. The cascade is modulated by feedback loops that include Plg activator inhibitor 1 (PAI-1). Molecular interactions with Plg kringle domains play an important role in regulating plasmin production and its modulation of fibrinolysis. We hypothesized that interactions of tissue plasminogen activator (tPA) with Plg kringle domains regulates plasmin levels in patients with stable CAD. METHODS: Plasma was collected from patients (n = 33) with an angiographically significant CAD and controls (n = 18) with angiographically established normal or minimally diseased arteries. Plasmin activity, tPA activity, and plasma levels of Plg, PAI-1, uPA, and tPA were determined. RESULTS: CAD patients had 1.7-fold greater plasmin activity (P = 0.02) that correlated with 1.5-fold higher tPA activity when compared to controls. Epitope mapping of Plg domains showed Plg differences in epitope exposure between the two groups. Plasma from CAD patients had 50% less (P < 0.001) detectable kringle 4 and 48% less (P = 0.007) detectable kringles 1-3. CONCLUSIONS: Based on detectable differences in Plg, we conclude that in patients with stable CAD, Plg complexed with tPA exists in a conformation that enables increased tPA activity and Plg conversion to plasmin. PMID- 16706974 TI - Circulating endothelial microparticles in acute ischemic stroke: a link to severity, lesion volume and outcome. AB - BACKGROUND: Endothelial membrane microparticles (EMP) in plasma are elevated in several vascular diseases. OBJECTIVES: To test the hypothesis that EMP would be increased in patients with acute ischemic stroke and would correlate with stroke severity, brain lesion volume and outcome. PATIENTS AND METHODS: Forty-one patients were studied and divided into two groups based on the National Institutes of Health Stroke Scale (NIHSS) score: 20 patients with mild stroke (NIHSS score < 5) and 21 patients with moderate-severe stroke (NIHSS score > or = 5). Lesion volume was measured using diffusion-weighted magnetic resonance imaging and discharge outcome was based on the discharge Barthel and Rankin scores. Twenty-three age-matched control subjects were also studied. Using flow cytometry, endoglin-positive EMP: CD105+ CD41a-CD45- (E(+)EMP), specific endothelial EMP expressing VE-cadherin and endoglin: CD105+CD144+ (C(+)EMP), EMP expressing phosphatidylserine: CD105+PS+ CD41a- (PS(+)EMP) and EMP expressing ICAM-1: CD105+CD54+ CD45- (I(+)EMP) were analyzed. RESULTS: Significantly higher PS(+)EMP counts were observed in the group of acute ischemic stroke patients [median 59 (25th-75th percentile: 28-86) MP microL(-1)] relative to the controls [28 (14-36) MP microL(-1)] (P = 0.002). All four EMP phenotypes studied were elevated in the subgroup of moderate-severe stroke patients relative to the controls (all P < 0.05). In the patients with acute ischemic stroke three EMP phenotypes (E(+)EMP, PS(+)EMP and I(+)EMP) correlated significantly with brain lesion volume, with I(+)EMP (P = 0.002) showing the strongest correlation. Admission counts of C(+)EMP (P = 0.0003) and E(+)EMP (P = 0.003) correlated significantly with discharge clinical outcome. CONCLUSIONS: Certain circulating EMP phenotypes may be associated with severity, lesion volume and outcome of acute ischemic stroke. EMP analysis shows promising contribution to understanding stroke pathophysiology. PMID- 16706975 TI - A genome-wide linkage scan for homocysteine levels suggests three regions of interest. AB - BACKGROUND: An elevated plasma total homocysteine (tHcy) level is a risk factor for many clinical conditions, including vascular disease and venous thrombosis. The tHcy levels are partly determined by genetic factors. Extensive candidate gene studies have identified several genetic variants, including the MTHFR 677C>T, that influence tHcy levels, but so far only part of the genetic variation in tHcy can be explained. OBJECTIVE: In order to identify chromosomal regions that influence tHcy levels, a genome-wide linkage analysis was conducted. PATIENTS/METHODS: Our study population consisted of 13 pedigrees and 469 subjects with data on fasting plasma tHcy levels. A set of 377 markers covering the genome was genotyped in 275 subjects. The variance component linkage method (SOLAR version 2.1.3) was used for the two-point and multipoint linkage analyses. RESULTS: The heritability of the age- and sex-adjusted homocysteine levels was 44%. The multipoint linkage analysis identified one region with suggestive linkage on chromosome 16q (LOD score 1.76; nominal P = 0.0024). Weaker evidence of linkage was found for regions on chromosome 12q (LOD score 1.57; nominal P = 0.0036) and chromosome 13q (LOD score 1.52; nominal P = 0.0041). CONCLUSIONS: In our families the plasma tHcy level was highly heritable. The multipoint linkage analysis identified three regions that showed weak to suggestive linkage to tHcy levels. PMID- 16706976 TI - Intracellular readthrough of nonsense mutations by aminoglycosides in coagulation factor VII. AB - BACKGROUND: Nonsense mutations in coagulation factor (F) VII potentially cause a lethal hemorrhagic diathesis. Readthrough of nonsense mutations by aminoglycosides has been studied in a few human disease models with variable results. OBJECTIVES: We investigated the K316X and W364X FVII mutations, associated with intracranial hemorrhage, and their correction by aminoglycosides. The rare nonsense mutations in FVII represent favorite models to test this strategy, because even tiny increases in the amount of functional full-length protein in patients could ameliorate hemorrhagic phenotypes. RESULTS: A FVII green fluorescent protein (GFP) chimaera provided us with a fluorescent model of FVII expression in living cells. Appreciable fluorescence in cells transfected with nonsense FVII-GFP mutants was detected upon geneticin treatment, thus demonstrating suppression of premature translation termination. To investigate the rescue of FVII function, nonsense variants of the native FVII without GFP (p316X-FVII and p364X-FVII) were transfected and found to secrete low amounts of FVII (approximately 1% of Wt-FVII activity), thus suggesting a spontaneous stop codon readthrough. Geneticin treatment of cells resulted in a significant and dose-dependent increase of secreted FVII molecules (p316X-FVII, 24 +/- 12 ng mL( 1), 3.6 +/- 0.8% of Wt-FVII activity; p364X-FVII, 26 +/- 10 ng mL(-1), 3.7+/ 0.6%) characterized by reduced specific activity, thus indicating the synthesis of dysfunctional proteins. Similar results were observed with gentamicin, a commonly used aminoglycoside of potential interest for patient treatment. CONCLUSIONS: Our approach, extendable to other coagulation factors, represents an effective tool for a systematic study of the effects of aminoglycosides and neighboring sequences on nonsense codon readthrough. These results provide the rationale for a mutation-specific therapeutic approach in FVII deficiency. PMID- 16706977 TI - Intrinsic stability and functional properties of disulfide bond-stabilized coagulation factor VIIIa variants. AB - BACKGROUND: The utility of purified coagulation factor (F)VIII for treatment of hemophilia A is limited in part by its instability following activation by thrombin, which is caused by spontaneous dissociation of the A2 domain from the activated FVIII (FVIIIa) heterotrimer. To prevent this A2 domain dissociation in FVIIIa, we previously engineered a cysteine pair (C664-C1826) in recombinant FVIII that formed a disulfide bond cross-linking the A2 domain in the heavy chain to the A3 domain in the light chain. This engineered disulfide bond resulted in a more stable FVIIIa. AIMS: Here, we characterize the functional parameters of C664 C1828 FVIII and of a new disulfide bond-stabilized FVIII (C662-C1828 FVIII). METHODS: In order to assess whether these FVIII variants might be good candidates for a new therapeutic agent to treat hemophilia A, we investigated a variety of functional parameters that might affect the in vivo properties of the variants, including half-life of disulfide bond-stabilized FVIII and FVIIIa and the potency of these FVIIIa molecules in the FXase complex. RESULTS: Both disulfide bond stabilized variants had improved affinity for von Willebrand factor (VWF). In studies of FX activation by purified FIXa and FVIIIa, C662-C1828 FVIIIa had normal activity while C664-C1826 FVIIIa had reduced activity. Both C664-C1826 FVIIIa and C662-C1828 FVIIIa were inactivated by activated protein C (APC) but the rates of inactivation were different. CONCLUSION: Overall, the specific location of the disulfide bridge between the A2 and A3 domains appears to affect functional properties of FVIIIa. In summary, introduction of engineered interdomain disulfides results in FVIIIa variants that resist spontaneous loss of activity while retaining susceptibility to APC proteolytic inactivation and maintaining VWF binding. PMID- 16706978 TI - Human polymorphonuclear leukocytes produce and express functional tissue factor upon stimulation. AB - BACKGROUND: Blood-borne tissue factor (TF) plays a crucial role in thrombogenesis. AIM: To study whether polymorphonuclear leukocytes (PMN) are a source of TF. METHODS AND RESULTS: Human PMN were carefully separated from other blood cells and stimulated for 3 min with purified P-selectin or the chemotactic peptide formyl-MetLeuPhe (fMLP): they expressed both TF procoagulant activity, as identified by specific TF MoAb and inactivated factor VIIa blockade; and TF:Ag (four to six times), as shown by flow-cytometry and immunocytochemistry. About 40% of permeabilized PMN, both resting and stimulated, contained TF:Ag, indicating that stimulation only modifies the location of TF:Ag within PMN. By real time-polymerase chain reaction (RT-PCR), a very low amount of TF mRNA was detectable in resting PMN, but a 3- to 5-fold increase was observed after 1-h stimulation with P-selectin or fMLP, respectively. CONCLUSIONS: These findings suggest that TF is not constitutively expressed in peripheral PMN, but can be up regulated and produced upon stimulation and specific gene transcription, as for instance during contact with activated platelets or endothelium. The stored TF is rapidly expressed in vitro as a functional molecule on the surface of activated PMN. The availability of PMN TF supports the relevance of inflammatory cells and their interaction with platelets for fibrin deposition and thrombus formation. PMID- 16706979 TI - The limits of simulation of the clotting system. AB - OBJECTIVE: To investigate in how far successful simulation of a thrombin generation (TG) curve gives information about the underlying biochemical reaction mechanism. RESULTS: The large majority of TG curves do not contain more information than can be expressed by four parameters. A limited kinetic mechanism of six reactions, comprising proteolytic activation of factor (F) X and FII, feedback activation of FV, a cofactor function of FVa and thrombin inactivation by antithrombin can simulate any TG curve in a number of different ways. The information content of a TG curve is thus much smaller than the information required to describe a physiologically realistic reaction scheme of TG. Consequently, much of the input information is irrelevant for the output. FVIII deficiency or activation of protein C can, for example, be simulated by a reaction mechanism in which these factors do not occur. CONCLUSION: A model that comprises not more than six reactions can simulate the same TG curve in a number of possible ways. The possibilities increase exponentially as the model grows more realistic. Successful simulation of experimental data therefore does not validate the underlying assumptions. A fortiori, simulation that is not checked against experimental data lacks any probative force. Simulation can be of use, however, to detect mistaken hypotheses and for parameter estimation in systems with fewer than five free parameters. PMID- 16706980 TI - International collaborative study for the calibration of a proposed international standard for thromboplastin, rabbit, plain. AB - BACKGROUND: A preparation of rabbit brain thromboplastin, provisionally coded 04/162, is proposed as a candidate for the World Health Organization (WHO) International Standard (IS) for thromboplastin (rabbit, plain), meant to replace the IS coded RBT/90 (rabbit, plain), stocks of which are now exhausted. RESULTS: The preparation was calibrated in an international collaborative study involving 21 laboratories from 13 countries and the calibration was performed against the existing WHO-IS (i.e. rTF/95 and OBT/79) and other Certified Reference Materials from the Institute for Reference Materials and Measurements of the European Commission (i.e. CRM149 S) and from the European Action on Anticoagulation (i.e. EUTHR-01). An additional candidate rabbit brain thromboplastin coded as 04/106 was also included in the study. On the basis of predefined criteria (the within- and between-laboratory precision of the calibration and the conformity to the calibration model), 04/162 was the preferred candidate. CONCLUSIONS: The assigned International Sensitivity Index value was 1.15 and the inter-laboratory SD and coefficient of variation were 0.057% and 4.9%, respectively. PMID- 16706981 TI - Molecular evolution caught in action: gene duplication and evolution of molecular isoforms of prothrombin activators in Pseudonaja textilis (brown snake). AB - BACKGROUND: The evolution of structurally and functionally similar proteins with highly diverse physiological roles within a single organism is of great interest. Australian elapid snakes offer an excellent opportunity to study the molecular evolution of prothrombin activators. Venom from Pseudonaja textilis contains pseutarin C, a group C prothrombin activator. Its enzymatic subunit is structurally and functionally similar to mammalian factor (F) Xa, whereas its non enzymatic subunit is similar to FVa. As vertebrates, the snakes also contain a system to activate prothrombin in their own blood during injury. These hemostatic factors are produced in the liver. RESULTS: Here we describe the presence of two molecular forms of FX expressed in the liver of P. textilis. Both isoforms have molecular signatures and domain architecture of FX. However, one isoform shows approximately 94% sequence identity with the snake FX from Tropidechis carinatus, whereas the other is much closer (90% identity) to the catalytic subunit of pseutarin C (PCCS). Real-time polymerase chain reaction reveals that the latter isoform is expressed approximately 56 000 times lower in the liver of P. textilis. However, the isoforms are not expressed in the venom gland. CONCLUSION: A detailed analysis of deletions and insertions along with the sequence indicates that the second isoform is an intermediate caught in the evolution of venom prothrombin activator from the blood coagulation FX. Thus, this isoform represents a 'molecular fossil' and reveals the likely evolutionary path of recruitment of FX in the venom gland. PMID- 16706982 TI - Factor V C2 domain contains a major thrombin-binding site responsible for thrombin-catalyzed factor V activation. AB - Factor (F)V is converted into its active form, FVa, by limited proteolysis. Thrombin-catalyzed activation of FV is essential for its full cofactor activation. Previously, we reported that thrombin was bound to the C2 domain in the light chain of FVIII. As FV has a similar domain structure to FVIII, we focused on the FV C2 domain as a possible binding region for thrombin. Kinetic parameters, measured by surface plasmon resonance, revealed that the K(d) values of anhydro-thrombin for FV, FVa, and the FV C2 domain were 66, 240, and 670 nmol L(-1), respectively. FV activation was increased by approximately 9-fold by the addition of thrombin. In the presence of the FV C2 domain, this increase of the FV activation was inhibited. However, FV activation was not inhibited by the addition of the FVIII C2 domain. FV was cleaved into a 105-kDa heavy chain and a 71/74-kDa light chain by thrombin-catalyzed proteolysis at Arg709, Arg1018 and Arg1545. In the presence of the FV C2 domain, the cleavage was inhibited at all sites. Proteolysis was not affected by the addition of the FVIII C2 domain. These results indicated that the FV C2 domain contains a major binding site for thrombin and that this domain is necessary for the proteolysis at all cleavage sites. Furthermore, the present results also suggested that thrombin has an independent binding site for FV different from that for FVIII. PMID- 16706983 TI - Increase in plasma plasminogen activators inhibitor type 1 concentration after fibrinolytic treatment in patients with acute myocardial infarction is associated with 4G/5G polymorphism of PAI-1 gene. AB - BACKGROUND: Preliminary data suggest that plasma concentration of plasminogen activators inhibitor type 1 (PAI-1) is genetically determined and may be related to differential regulation of plasma PAI-1 concentration at baseline and after stimulation. AIM: This study aimed to evaluate whether increase in the plasma PAI 1 antigen concentration or activity after fibrinolytic therapy in patients with acute myocardial infarction is associated with the -675 4G/5G genetic polymorphism in the promoter region of PAI-1 gene. RESULTS & CONCLUSIONS: Our study revealed that a rebound effect is observed in PAI-1 activity (ActPAI-1) and PAI-1 antigen (AgPAI-1) concentration after standard streptokinase treatment with maximal values of 3 h (t3) after the completion of streptokinase infusion. Both ActPAI-1 and AgPAI-1 were significantly higher at t3 compared to the levels before fibrinolytic treatment: 37.3 (20.0-67.7) vs. 10.0 (3.6-26.0) IU L(-1); P = 0.00001 and 29.9 (15.6-42.3) vs. 20.9 (13.0-30.2) ng mL(-1); P = 0.001, respectively. The stratification of the patients by genotype revealed that carriers of the 4G allele had higher concentrations of PAI-1 antigen 3 h after streptokinase infusion: 30.9 vs. 13.8 ng mL(-1); P = 0.019. No significant association between PAI-1 activity and genotype was found. In conclusion, the rebound effect in serum PAI-1 concentration observed after streptokinase treatment may be related to the 4G/5G polymorphism in the PAI-1 gene promoter. PMID- 16706984 TI - von Willebrand factor mediates platelet spreading through glycoprotein Ib and alpha(IIb)beta3 in the presence of botrocetin and ristocetin, respectively. AB - BACKGROUND: von Willebrand factor (VWF) plays a critical role in the process of hemostasis by mediating flow-dependent adhesion and spreading of platelets on exposed extracellular matrix proteins following vascular injury. To accomplish this, VWF binds to two distinct platelet receptors: glycoprotein (GP)Ib-IX-V and integrin alpha(IIb)beta3. OBJECTIVE: To evaluate the ability of GPIb and alpha(IIb)beta3 to mediate platelet adhesion and lamellipodia formation on immobilized VWF in the presence of the biochemical modulators, ristocetin and botrocetin. RESULTS: In the presence of botrocetin and inhibitors of adenosine diphosphate (ADP) and thromboxane A2 (TxA2), VWF is able to support formation of lamellipodia through a GPIb-dependent mechanism that is independent of alpha(IIb)beta3 and PI3-kinase. Lamellipodia formation under these conditions is incomplete. In marked contrast, in the presence of ristocetin, VWF stimulates formation of fully spread lamellipodia through a pathway that is dependent upon alpha(IIb)beta3 and PI3-kinase. Furthermore, alpha(IIb)beta3 also supports platelet spreading on VWF alone, but only in the absence of inhibitors of ADP and TxA2. The localization of filamentous actin and the Arp2/3 complex in platelets on VWF in the presence of botrocetin and ristocetin are distinct, yielding disparate lamellipodium kinetic signatures. Interestingly, botrocetin significantly enhances platelet adhesion to VWF under flow in whole blood in an alpha(IIb)beta3-independent manner, while ristocetin augments washed platelet adhesion and spreading to VWF under flow in an alpha(IIb)beta3-dependent manner. CONCLUSIONS: These observations demonstrate that VWF is able to induce lamellipodia formation through distinct receptors, and has important consequences for investigation of the role of VWF-GPIb interactions in the context of platelet regulation. PMID- 16706985 TI - Critical role of ADP interaction with P2Y12 receptor in the maintenance of alpha(IIb)beta3 activation: association with Rap1B activation. AB - OBJECTIVE: Platelet integrin alpha(IIb)beta3 plays a crucial role in platelet aggregation, and the affinity of alpha(IIb)beta3 for fibrinogen is dynamically regulated. Employing modified ligand-binding assays, we analyzed the mechanism by which alpha(IIb)beta3 maintains its high-affinity state. METHODS AND RESULTS: Washed platelets adjusted to 50 x 10(3) microL(-1) were stimulated with 0.2 U mL( 1) thrombin or 5 microm U46619 under static conditions. After the completion of alpha(IIb)beta3 activation and granule secretion, different kinds of antagonists were added to the activated platelets. The activated alpha(IIb)beta3 was then detected by fluorescein isothiocyanate (FITC)-labeled PAC1. The addition of 1 mum AR-C69931MX (a P2Y12 antagonist) or 1 mm A3P5P (a P2Y1 antagonist) disrupted the sustained alpha(IIb)beta3 activation by approximately 92% and approximately 38%, respectively, without inhibiting CD62P or CD63 expression. Dilution of the platelet preparation to 500 microL(-1) also disrupted the sustained alpha(IIb)beta3 activation, and the disruption by such dilution was abrogated by the addition of exogenous adenosine 5'-diphosphate (ADP) in a dose-dependent fashion. The amounts of ADP released from activated platelets determined by high performance liquid chromatography were compatible with the amounts of exogenous ADP required for the restoration. We next examined the effects of antagonists on protein kinase C (PKC) and Rap1B activation induced by 0.2 U mL(-1) thrombin. Thrombin induced long-lasting PKC and Rap1B activation. AR-C69931MX markedly inhibited Rap1B activation without inhibiting PKC activation. CONCLUSIONS: Our data indicate that the continuous interaction between released ADP and P2Y12 is critical for the maintenance of alpha(IIb)beta3 activation. PMID- 16706987 TI - Human endothelial cells synthesize and release ADAMTS-13. AB - Hepatic stellate cells have been considered to be a primary source for human plasma ADAMTS-13, the von Willebrand factor (VWF)-cleaving metalloprotease. In this study, ADAMTS-13 antigen was detected by immunofluorescence in both venous (HUVECs) and arterial endothelial cells (HUAECs) using both polyclonal antibodies made against peptides found in various domains of human ADAMTS-13, as well as by a monoclonal antibody against the ADAMTS-13 metalloprotease domain. ADAMTS-13 antigen had an intra-cellular distribution in endothelial cells distinct from the Weibel-Palade body location of VWF, and was released from the cells during 48 h in culture. The mRNA for ADAMTS13 was detected in HUVECs and HUAECs using reverse transcription-polymerase chain reaction (RT-PCR), indicating that the enzyme is synthesized in these cells. The ADAMTS-13 protein was immunoprecipitated from HUVECs and had an approximate M(r) of 170 kDa, similar to the molecular mass of recombinant ADAMTS-13. The ADAMTS-13 in HUVEC and HUAEC lysates had enzymatic activity using both static and flow assays. We conclude that ADAMTS-13 is synthesized in human endothelial cells, and released constitutively. The vast number of endothelial cells in the body may be an important source of ADAMTS-13. PMID- 16706986 TI - Platelet septin complexes form rings and associate with the microtubular network. AB - BACKGROUND: Septins are important effectors in molecular mechanisms involving membrane partitioning. To date, a growing repertoire of septins in mammals includes 13 different proteins (SEPT1 to SEPT13) that can be classified into four distinct categories based on sequence similarity. AIM: In this study, we document the human platelet septin, SEPT5, as part of a complex composed of multiple septin proteins. RESULTS: Biochemical and immunofluorescent data place the majority of these complexes in the platelet periphery as part of the platelet circumferential band copurifying with the platelet microtubule coil and tubulin. The presence of a prominent platelet septin ring in resting platelets appears to be left intact in the activated platelet, as a similar ring structure is observed following platelet spreading on fibrinogen. The ablation of SEPT5 in the knock out mouse model had previously been reported to result in a platelet phenotype with aggregation using subthreshold levels of agonist. Speculation on the role of SEPT5 in the platelet-release reaction suggested that SEPT5 regulates platelet function by association with platelet storage granules. We now report that the absence of SEPT5 results in increased ATP release from stimulated platelets. CONCLUSION: These studies document the presence of platelet septin complexes and validate the importance of septins for platelet physiology. PMID- 16706988 TI - Patterns of changes of anti-ADAMTS13 after plasma exchange. PMID- 16706989 TI - Activated partial thromboplastin time: results of a case-control study evaluating six commercial reagents in assessing the risk of venous thromboembolism. PMID- 16706990 TI - Factor Xa protects cultured hippocampal neurons from glutamate toxicity. PMID- 16706991 TI - Low incidence of venographically detected deep vein thrombosis after knee arthroscopy without thromboprophylaxis: a prospective cohort study. PMID- 16706992 TI - Beta-thalassemia intermedia and pregnancy: should we anticoagulate? PMID- 16706993 TI - Regulation of endogenous tissue-type plasminogen activator expression is modulated by the -7351C>T enhancer polymorphism. PMID- 16706994 TI - The relationship between ABO blood group and the risk of bleeding during vitamin K antagonist treatment. PMID- 16706995 TI - Extra-virgin vs. refined olive oil on postprandial hemostatic markers in healthy subjects. PMID- 16706996 TI - The impact of simvastatin on warfarin disposition and dose requirements. PMID- 16706997 TI - Can the PFA-100 be modified to detect P2Y12 inhibition? PMID- 16706998 TI - More on: platelet function analyzer (PFA)-100 closure time in the evaluation of platelet disorders and platelet function. PMID- 16707001 TI - The reactivity of paired plasma and serum samples are comparable in the anticardiolipin and anti-beta2-glycoprotein-1 ELISAs: a rebuttal. PMID- 16707004 TI - The success (or not) of HUGO nomenclature. AB - Current usage of gene nomenclature is ambiguous and impairs the efficient handling of scientific information. Therefore it is important to propose guidelines to deal with this problem. This study attempts to evaluate the success of HUGO nomenclature for human genes. The results indicate that HUGO guidelines are not supported by the scientific community. PMID- 16707005 TI - Non-specific low back pain in primary care in the Spanish National Health Service: a prospective study on clinical outcomes and determinants of management. AB - BACKGROUND: The Spanish National Health Service is a universal and free health care system. Non-specific low back pain (LBP) is a prevalent disorder, generating large health and social costs. The objectives of this study were to describe its management in primary care, to assess patient characteristics that influence physicians' decisions, and to describe clinical outcome at 2 months. METHODS: A cross-sectional sample of 648 patients with non-specific low back pain was recruited by 75 physicians (out of 361-20.8%) working in 40 primary care centers in 10 of the 17 administrative regions in Spain, covering 693,026 out of the 40,499,792 inhabitants. Patients were assessed on the day they were recruited, and prospectively followed-up 14 and 60 days later. The principal patient characteristics that were analyzed were: sex, duration of the episode, history of LBP, working status, severity of LBP, leg pain and disability, and results of straight leg raising test. Descriptors of management were: performance of the straight leg raising test, ordering of diagnostic procedures, prescription of drug treatment, referral to physical therapy, rehabilitation or surgery, and granting of sick leave. Regression analysis was used to analyze the relationship between patients' baseline characteristics and physicians' management decisions. Only workers were included in the models on sick leave. RESULTS: Mean age (SD) of included patients was 46.5 (15.5) years, 367 (56.6%) were workers, and 338 (52.5%) were females. Median (25th-75th interquartile range) duration of pain when entering the study was 4 (2-10) days and only 28 patients (4.3%) had chronic low back pain. Diagnostic studies included plain radiographs in 43.1% of patients and CT or MRI scans in 18.8%. Drug medication was prescribed to 91.7% of patients, 19.1% were sent to physical therapy or rehabilitation, and 9.6% were referred to surgery. The main determinants of the clinical management were duration of the episode and, to a lesser extent, the intensity of the pain (especially leg pain), a positive straight leg raising test, and degree of disability. The main determinant of sick leave was the degree of disability, followed by the characteristics of the labor contract and the intensity of leg pain (but not low back pain). After at least 2 months of treatment, 37% of patients were still in pain and approximately 10% of patients had not improved or had worsened. CONCLUSION: Although the use of X-Rays is high, determinants of physicians' management of LBP in primary care made clinical sense and were consistent with patterns suggested by evidence-based recommendations. However, after 2 months of treatment more than one third of patients continued to have back pain and about 10% had worsened. PMID- 16707006 TI - The development and validation of the Virtual Tissue Matrix, a software application that facilitates the review of tissue microarrays on line. AB - BACKGROUND: The Tissue Microarray (TMA) facilitates high-throughput analysis of hundreds of tissue specimens simultaneously. However, bottlenecks in the storage and manipulation of the data generated from TMA reviews have become apparent. A number of software applications have been developed to assist in image and data management; however no solution currently facilitates the easy online review, scoring and subsequent storage of images and data associated with TMA experimentation. RESULTS: This paper describes the design, development and validation of the Virtual Tissue Matrix (VTM). Through an intuitive HTML driven user interface, the VTM provides digital/virtual slide based images of each TMA core and a means to record observations on each TMA spot. Data generated from a TMA review is stored in an associated relational database, which facilitates the use of flexible scoring forms. The system allows multiple users to record their interpretation of each TMA spot for any parameters assessed. Images generated for the VTM were captured using a standard background lighting intensity and corrective algorithms were applied to each image to eliminate any background lighting hue inconsistencies or vignetting. Validation of the VTM involved examination of inter-and intra-observer variability between microscope and digital TMA reviews. Six bladder TMAs were immunohistochemically stained for E Cadherin, beta-Catenin and PhosphoMet and were assessed by two reviewers for the amount of core and tumour present, the amount and intensity of membrane, cytoplasmic and nuclear staining. CONCLUSION: Results show that digital VTM images are representative of the original tissue viewed with a microscope. There were equivalent levels of inter-and intra-observer agreement for five out of the eight parameters assessed. Results also suggest that digital reviews may correct potential problems experienced when reviewing TMAs using a microscope, for example, removal of background lighting variance and tint, and potential disorientation of the reviewer, which may have resulted in the discrepancies evident in the remaining three parameters. PMID- 16707007 TI - Frozen section is superior to imprint cytology for the intra-operative assessment of sentinel lymph node metastasis in stage I breast cancer patients. AB - BACKGROUND: A standard intra-operative procedure for assessing sentinel lymph node metastasis in breast cancer patients has not yet been established. PATIENTS AND METHODS: One hundred and thirty-eight patients with stage I breast cancer who underwent sentinel node biopsy using both imprint cytology and frozen section were analyzed. RESULTS: Seventeen of the 138 patients had sentinel node involvement. Results of imprint cytology included nine false negative cases (sensitivity, 47.1%). In contrast, only two cases of false negatives were found on frozen section (sensitivity, 88.2%). There were two false positive cases identified by imprint cytology (specificity, 98.3%). On the other hand, frozen section had 100% specificity. CONCLUSION: These findings suggest that frozen section is superior to imprint cytology for the intra-operative determination of sentinel lymph node metastasis in stage I breast cancer patients. PMID- 16707008 TI - Somatic VHL gene alterations in MEN2-associated medullary thyroid carcinoma. AB - BACKGROUND: Germline mutations in RET are responsible for multiple endocrine neoplasia type 2 (MEN2), an autosomal dominantly inherited cancer syndrome that is characterized by medullary thyroid carcinoma (MTC), pheochromocytoma, and parathyroid hyperplasia/adenoma. Recent studies suggest a "second hit" mechanism resulting in amplification of mutant RET. Somatic VHL gene alterations are implicated in the pathogenesis of MEN2 pheochromocytomas. We hypothesized that somatic VHL gene alterations are also important in the pathogenesis of MEN2 associated MTC. METHODS: We analyzed 6 MTCs and 1 C-cell hyperplasia (CCH) specimen from 7 patients with MEN2A and RET germline mutations in codons 609, 618, 620, or 634, using microdissection, microsatellite analysis, phosphorimage densitometry, and VHL mutation analysis. RESULTS: First, we searched for allelic imbalance between mutant and wild-type RET by using the polymorphic markers D10S677, D10S1239, and RET on thyroid tissue from these patients. Evidence for RET amplification by this technique could be demonstrated in 3 of 6 MTCs. We then performed LOH analysis using D3S1038 and D3S1110 which map to the VHL gene locus at 3p25/26. VHL gene deletion was present in 3 MTCs. These 3 MTCs also had an allelic imbalance between mutant and wild-type RET. Mutation analysis of the VHL gene showed a somatic frameshift mutation in 1 MTC that also demonstrated LOH at 3p25/26. In the 2 other MTCs with allelic imbalance of RET and somatic VHL gene deletion, no somatic VHL mutation could be detected. The CCH specimen did neither reveal RET imbalance nor somatic VHL gene alterations. CONCLUSION: These data suggest that a RET germline mutation is necessary for development of CCH, that allelic imbalance between mutant and wild-type RET may set off tumorigenesis, and that somatic VHL gene alterations may not play a major role in tumorigenesis of MEN2A-associated MTC. PMID- 16707009 TI - Teenage suicide cluster formation and contagion: implications for primary care. AB - BACKGROUND: We have previously studied unintentional as well as intentional injury deaths among teenagers living in the four northernmost counties, forming approximately 55% of Sweden with 908,000 inhabitants in 1991. During this work, we found what we suspected to be a suicide cluster among teenagers and we also suspected contagion since there were links between these cases. In this present study, we investigate the occurrence of suicide clustering among teenagers, analyze cluster definitions, and suggest preventive measures. METHODS: A retrospective study of teenager suicides autopsied at the Department of Forensic Medicine in Umea, Sweden, during 1981 through 2000. Police reports, autopsy protocols, and medical records were studied in all cases, and the police officers that conducted the investigation at the scene were interviewed in all cluster cases. Parents of the suicide victims of the first cluster were also interviewed. Two aggregations of teenager suicides were detected and evaluated as possible suicide clusters using the US Centers for Disease Control definition of a suicide cluster. RESULTS: Two clusters including six teenagers were confirmed, and contagion was established within each cluster. CONCLUSION: The general practitioner is identified as a key person in the aftermath of a teenage suicide since the general practitioner often meet the family, friends of the deceased, and other acquaintances early in the process after a suicide. This makes the general practitioner suitable to initiate contacts with others involved in the well-being of the young, in order to prevent suicide cluster formation and para suicidal activities. PMID- 16707010 TI - On the front line of primary health care: the profile of community health workers in rural Quechua communities in Peru. AB - OBJECTIVE: To describe the profile of community health workers--health promoters, traditional birth attendants and traditional healers--in rural Quechua communities from Ayacucho, Peru. METHODS: Basic quantitative and qualitative information was gathered as part of a community health project implemented between 1997 and 2002 in 40 Andean communities with information from questionnaires, personal interviews and group discussions. RESULTS: The majority of current community health workers are men with limited education who are primarily Quechua speakers undertaking their work on a voluntary basis. Health promoters are mostly young, male, high school graduates. There exists a high drop out rate among these workers. In contrast, traditional healers and traditional birth attendants possess an almost diametrically opposite profile in terms of age, education and drop-out rates, though males still predominate. At the community level the health promoters are the most visible community health workers. CONCLUSION: It is very important to consider and to be aware of the profile of community health workers in order to provide appropriate alternatives when working with these groups as well as with the indigenous population, particularly in terms of culture, language and gender issues. PMID- 16707011 TI - Increasing participation of cancer patients in randomised controlled trials: a systematic review. AB - BACKGROUND: There are many barriers to patient participation in randomised controlled trials of cancer treatments. To increase participation in trials, strategies need to be identified to overcome these barriers. Our aim was to assess the effectiveness of interventions to overcome barriers to patient participation in randomised controlled trials (RCTs) of cancer treatments. METHODS: A systematic review was conducted. Published and unpublished studies in any language were searched for in fifteen electronic databases, including MEDLINE, EMBASE, CINAHL and PsycINFO, from inception to the end of 2004. Studies of any interventions to improve cancer patient participation in RCTs, which reported the change in recruitment rates, were eligible for inclusion. RCTs and non-randomised controlled trials as well as before and after studies reporting baseline rates specific to the population being investigated were included. Data were extracted by one reviewer into structured summary tables and checked for accuracy by a second reviewer. Each included study was assessed against a checklist for methodological quality by one reviewer and checked by a second reviewer. A narrative synthesis was conducted. RESULTS: Eight studies were identified that met the inclusion criteria: three RCTs, two non-randomised controlled trials and three observational studies. Six of the studies had an intervention that had some relevance to the UK. There was no robust evidence that any of the interventions investigated led to an increase in cancer patient participation in RCTs, though one good quality RCT found that urologists and nurses were equally effective at recruiting participants to a treatment trial for prostate cancer. Although there was no evidence of an effect in any of the studies, the evidence was not of sufficient quality to be able to conclude that these interventions therefore do not work. CONCLUSION: There is not a strong evidence-base for interventions that increase cancer patient participation in randomised trials. Further research is required to evaluate the effectiveness of strategies to increase participation in cancer treatment trials. PMID- 16707012 TI - Tuberculosis contact investigation with a new, specific blood test in a low incidence population containing a high proportion of BCG-vaccinated persons. AB - BACKGROUND: BCG-vaccination can confound tuberculin skin test (TST) reactions in the diagnosis of latent tuberculosis infection. METHODS: We compared the TST with a Mycobacterium tuberculosis specific whole blood interferon-gamma assay (QuantiFERON-TB-Gold In Tube; QFT-G) during ongoing investigations among close contacts of sputum smear positive source cases in Hamburg, Germany. RESULTS: During a 6-month period, 309 contacts (mean age 28.5 +/- 10.5 years) from a total of 15 source cases underwent both TST and QFT-G testing. Of those, 157 (50.8%) had received BCG vaccination and 84 (27.2%) had migrated to Germany from a total of 25 different high prevalence countries (i.e. >20 cases/100,000). For the TST, the positive response rate was 44.3% (137/309), whilst only 31 (10%) showed a positive QFT-G result. The overall agreement between the TST and the QFT-G was low (kappa = 0.2, with 95% CI 0.14.-0.23), and positive TST reactions were closely associated with prior BCG vaccination (OR 24.7; 95% CI 11.7-52.5). In contrast, there was good agreement between TST and QFT-G in non-vaccinated persons (kappa = 0.58, with 95% CI 0.4-0.68), increasing to 0.68 (95% CI 0.46 0.81), if a 10-mm cut off for the TST was used instead of the standard 5 mm recommended in Germany. CONCLUSION: The QFT-G assay was unaffected by BCG vaccination status, unlike the TST. In close contacts who were BCG-vaccinated, the QFT-G assay appeared to be a more specific indicator of latent tuberculosis infection than the TST, and similarly sensitive in unvaccinated contacts. In BCG vaccinated close contacts, measurement of IFN-gamma responses of lymphocytes stimulated with M. tuberculosis-specific antigen should be recommended as a basis for the decision on whether to perform subsequent chest X-ray examinations or to start treatment for latent tuberculosis infection. PMID- 16707013 TI - Immunization coverage and risk factors for failure to immunize within the Expanded Programme on Immunization in Kenya after introduction of new Haemophilus influenzae type b and hepatitis b virus antigens. AB - BACKGROUND: Kenya introduced a pentavalent vaccine including the DTP, Haemophilus influenzae type b and hepatitis b virus antigens in Nov 2001 and strengthened immunization services. We estimated immunization coverage before and after introduction, timeliness of vaccination and risk factors for failure to immunize in Kilifi district, Kenya. METHODS: In Nov 2002 we performed WHO cluster-sample surveys of >200 children scheduled for vaccination before or after introduction of pentavalent vaccine. In Mar 2004 we conducted a simple random sample (SRS) survey of 204 children aged 9-23 months. Coverage was estimated by inverse Kaplan Meier survival analysis of vaccine-card and mothers' recall data and corroborated by reviewing administrative records from national and provincial vaccine stores. The contribution to timely immunization of distance from clinic, seasonal rainfall, mother's age, and family size was estimated by a proportional hazards model. RESULTS: Immunization coverage for three DTP and pentavalent doses was 100% before and 91% after pentavalent vaccine introduction, respectively. By SRS survey, coverage was 88% for three pentavalent doses. The median age at first, second and third vaccine dose was 8, 13 and 18 weeks. Vials dispatched to Kilifi District during 2001-2003 would provide three immunizations for 92% of the birth cohort. Immunization rate ratios were reduced with every kilometre of distance from home to vaccine clinic (HR 0.95, CI 0.91-1.00), rainy seasons (HR 0.73, 95% CI 0.61-0.89) and family size, increasing progressively up to 4 children (HR 0.55, 95% CI 0.41-0.73). CONCLUSION: Vaccine coverage was high before and after introduction of pentavalent vaccine, but most doses were given late. Coverage is limited by seasonal factors and family size. PMID- 16707014 TI - Stochastic modeling of oligodendrocyte generation in cell culture: model validation with time-lapse data. AB - BACKGROUND: The purpose of this paper is two-fold. The first objective is to validate the assumptions behind a stochastic model developed earlier by these authors to describe oligodendrocyte generation in cell culture. The second is to generate time-lapse data that may help biomathematicians to build stochastic models of cell proliferation and differentiation under other experimental scenarios. RESULTS: Using time-lapse video recording it is possible to follow the individual evolutions of different cells within each clone. This experimental technique is very laborious and cannot replace model-based quantitative inference from clonal data. However, it is unrivalled in validating the structure of a stochastic model intended to describe cell proliferation and differentiation at the clonal level. In this paper, such data are reported and analyzed for oligodendrocyte precursor cells cultured in vitro. CONCLUSION: The results strongly support the validity of the most basic assumptions underpinning the previously proposed model of oligodendrocyte development in cell culture. However, there are some discrepancies; the most important is that the contribution of progenitor cell death to cell kinetics in this experimental system has been underestimated. PMID- 16707015 TI - The wider social environment and changes in self-reported quality of life in the transition from late childhood to early adolescence: a cohort study. AB - BACKGROUND: Neighbourhood socioeconomic disadvantage and social capital have been associated with adolescent well-being, but the majority of studies were cross sectional, and the time window over which the neighbourhood may impact on development is unknown. Therefore, the contribution of the neighbourhood environment to adolescents' quality of life and the course of these effects during the period of transition from childhood to early adolescence was examined. METHODS: A cohort of adolescents living in Maastricht (The Netherlands), with a mean age of 11.2 years at baseline and of 13.5 years at follow-up was followed. Adolescents who responded both at baseline and at follow-up were included in the analysis (n = 475). Multilevel regression analyses estimated neighbourhood effects while controlling for individual-level effects. Neighbourhood-level socioeconomic and social capital variables, individual-level confounders, and baseline values of the outcome measures were included in the models. RESULTS: None of the neighbourhood factors was associated with changes in general health or mental health over the two-year period. However, two-year exposure to greater disparity between individual level socioeconomic status on the one hand and neighbourhood level of socioeconomic status on the other (e.g. high socioeconomic status adolescents living in deprived neighbourhoods and vice versa) negatively impacted on self-esteem and satisfaction. CONCLUSION: The neighbourhood environment per se does not contribute to change in quality of life during the transition to early adolescence. However, adolescents living in families whose socioeconomic status deviates from the mean level of neighbourhood socioeconomic deprivation may be negatively affected. PMID- 16707016 TI - Prostate response to prolactin in sexually active male rats. AB - BACKGROUND: The prostate is a key gland in the sexual physiology of male mammals. Its sensitivity to steroid hormones is widely known, but its response to prolactin is still poorly known. Previous studies have shown a correlation between sexual behaviour, prolactin release and prostate physiology. Thus, here we used the sexual behaviour of male rats as a model for studying this correlation. Hence, we developed experimental paradigms to determine the influence of prolactin on sexual behaviour and prostate organization of male rats. METHODS: In addition to sexual behaviour recordings, we developed the ELISA procedure to quantify the serum level of prolactin, and the hematoxilin-eosin technique for analysis of the histological organization of the prostate. Also, different experimental manipulations were carried out; they included pituitary grafts, and haloperidol and ovine prolactin treatments. Data were analyzed with a One way ANOVA followed by post hoc Dunnet test if required. RESULTS: Data showed that male prolactin has a basal level with two peaks at the light-dark-light transitions. Consecutive ejaculations increased serum prolactin after the first ejaculation, which reached the highest level after the second, and started to decrease after the third ejaculation. These normal levels of prolactin did not induce any change at the prostate tissue. However, treatments for constant elevations of serum prolactin decreased sexual potency and increased the weight of the gland, the alveoli area and the epithelial cell height. Treatments for transient elevation of serum prolactin did not affect the sexual behaviour of males, but triggered these significant effects mainly at the ventral prostate. CONCLUSION: The prostate is a sexual gland that responds to prolactin. Mating induced prolactin release is required during sexual encounters to activate the epithelial cells in the gland. Here we saw a precise mechanism controlling the release of prolactin during ejaculations that avoid the detrimental effects produced by constant levels. However, we showed that minor elevations of prolactin which do not affect the sexual behaviour of males, produced significant changes at the prostate epithelium that could account for triggering the development of hyperplasia or cancer. Thus, it is suggested that minute elevations of serum prolactin in healthy subjects are at the etiology of prostate abnormal growth. PMID- 16707017 TI - Violent crime exposure classification and adverse birth outcomes: a geographically-defined cohort study. AB - BACKGROUND: Area-level socioeconomic disparities have long been associated with adverse pregnancy outcomes. Crime is an important element of the neighborhood environment inadequately investigated in the reproductive and public health literature. When crime has been used in research, it has been variably defined, resulting in non-comparable associations across studies. METHODS: Using geocoded linked birth record, crime and census data in multilevel models, this paper explored the relevance of four spatial violent crime exposures: two proximal violent crime categorizations (count of violent crime within a one-half mile radius of maternal residence and distance from maternal residence to nearest violent crime) and two area-level crime categorizations (count of violent crimes within a block group and block group rate of violent crimes) for adverse birth events among women in living in the city of Raleigh NC crime report area in 1999 2001. Models were adjusted for maternal age and education and area-level deprivation. RESULTS: In black and white non-Hispanic race-stratified models, crime characterized as a proximal exposure was not able to distinguish between women experiencing adverse and women experiencing normal birth outcomes. Violent crime characterized as a neighborhood attribute was positively associated with preterm birth and low birth weight among non-Hispanic white and black women. No statistically significant interaction between area-deprivation and violent crime category was observed. CONCLUSION: Crime is variably categorized in the literature, with little rationale provided for crime type or categorization employed. This research represents the first time multiple crime categorizations have been directly compared in association with health outcomes. Finding an effect of area-level violent crime suggests crime may best be characterized as a neighborhood attribute with important implication for adverse birth outcomes. PMID- 16707018 TI - Analysis of folylpoly-gamma-glutamate synthetase gene expression in human B precursor ALL and T-lineage ALL cells. AB - BACKGROUND: Expression of folylpoly-gamma-glutamate synthetase (FPGS) gene is two to three-fold higher in B-precursor ALL (Bp- ALL) than in T-lineage ALL (T-ALL) and correlates with intracellular accumulation of methotrexate (MTX) polyglutamates and lymphoblast sensitivity to MTX. In this report, we investigated the molecular regulatory mechanisms directing FPGS gene expression in Bp-ALL and T-ALL cells. METHODS: To determine FPGS transcription rate in Bp ALL and T-ALL we used nuclear run-on assays. 5'-RACE was used to uncover potential regulatory regions involved in the lineage differences. We developed a luciferase reporter gene assay to investigate FPGS promoter/enhancer activity. To further characterize the FPGS proximal promoter, we determined the role of the putative transcription binding sites NFY and E-box on FPGS expression using luciferase reporter gene assays with substitution mutants and EMSA. RESULTS: FPGS transcription initiation rate was 1.6-fold higher in NALM6 vs. CCRF-CEM cells indicating that differences in transcription rate led to the observed lineage differences in FPGS expression between Bp-ALL and T-ALL blasts. Two major transcripts encoding the mitochondrial/cytosolic and cytosolic isoforms were detected in Bp-ALL (NALM6 and REH) whereas in T-ALL (CCRF-CEM) cells only the mitochondrial/cytosolic transcript was detected. In all DNA fragments examined for promoter/enhancer activity, we measured significantly lower luciferase activity in NALM6 vs. CCRF-CEM cells, suggesting the need for additional yet unidentified regulatory elements in Bp-ALL. Finally, we determined that the putative transcription factor binding site NFY, but not E-box, plays a role in FPGS transcription in both Bp- and T-lineage. CONCLUSION: We demonstrated that the minimal FPGS promoter region previously described in CCRF-CEM is not sufficient to effectively drive FPGS transcription in NALM6 cells, suggesting that different regulatory elements are required for FPGS gene expression in Bp cells. Our data indicate that the control of FPGS expression in human hematopoietic cells is complex and involves lineage-specific differences in regulatory elements, transcription initiation rates, and mRNA processing. Understanding the lineage-specific mechanisms of FPGS expression should lead to improved therapeutic strategies aimed at overcoming MTX resistance or inducing apoptosis in leukemic cells. PMID- 16707019 TI - Genetic structure of Indian populations based on fifteen autosomal microsatellite loci. AB - BACKGROUND: Indian populations endowed with unparalleled genetic complexity have received a great deal of attention from scientists world over. However, the fundamental question over their ancestry, whether they are all genetically similar or do exhibit differences attributable to ethnicity, language, geography or socio-cultural affiliation is still unresolved. In order to decipher their underlying genetic structure, we undertook a study on 3522 individuals belonging to 54 endogamous Indian populations representing all major ethnic, linguistic and geographic groups and assessed the genetic variation using autosomal microsatellite markers. RESULTS: The distribution of the most frequent allele was uniform across populations, revealing an underlying genetic similarity. Patterns of allele distribution suggestive of ethnic or geographic propinquity were discernible only in a few of the populations and was not applicable to the entire dataset while a number of the populations exhibited distinct identities evident from the occurrence of unique alleles in them. Genetic substructuring was detected among populations originating from northeastern and southern India reflective of their migrational histories and genetic isolation respectively. CONCLUSION: Our analyses based on autosomal microsatellite markers detected no evidence of general clustering of population groups based on ethnic, linguistic, geographic or socio-cultural affiliations. The existence of substructuring in populations from northeastern and southern India has notable implications for population genetic studies and forensic databases where broad grouping of populations based on such affiliations are frequently employed. PMID- 16707020 TI - angaGEDUCI: Anopheles gambiae gene expression database with integrated comparative algorithms for identifying conserved DNA motifs in promoter sequences. AB - BACKGROUND: The completed sequence of the Anopheles gambiae genome has enabled genome-wide analyses of gene expression and regulation in this principal vector of human malaria. These investigations have created a demand for efficient methods of cataloguing and analyzing the large quantities of data that have been produced. The organization of genome-wide data into one unified database makes possible the efficient identification of spatial and temporal patterns of gene expression, and by pairing these findings with comparative algorithms, may offer a tool to gain insight into the molecular mechanisms that regulate these expression patterns. DESCRIPTION: We provide a publicly-accessible database and integrated data-mining tool, angaGEDUCI, that unifies 1) stage- and tissue specific microarray analyses of gene expression in An. gambiae at different developmental stages and temporal separations following a bloodmeal, 2) functional gene annotation, 3) genomic sequence data, and 4) promoter sequence comparison algorithms. The database can be used to study genes expressed in particular stages, tissues, and patterns of interest, and to identify conserved promoter sequence motifs that may play a role in the regulation of such expression. The database is accessible from the address http://www.angaged.bio.uci.edu. CONCLUSION: By combining gene expression, function, and sequence data with integrated sequence comparison algorithms, angaGEDUCI streamlines spatial and temporal pattern-finding and produces a straightforward means of developing predictions and designing experiments to assess how gene expression may be controlled at the molecular level. PMID- 16707022 TI - Interaction of obesity, metabolic syndrome and Framingham risk on steatohepatitis among healthy Taiwanese: population-based nested case-control study. AB - BACKGROUND: There have been scant reports on the cumulative effects of atherosclerotic risk factors on steatohepatitis. METHODS: We defined cases of steatohepatitis (n = 124) from one health examination center at National Taiwan University Hospital from January to December 2002. We selected controls, matched by age, gender and drinking status. Metabolic syndrome was defined by the modified ATP-III guidelines. High-dimensional interactions of risk factors for steatohepatitis were evaluated. RESULTS: Steatohepatitis cases had the highest C reactive protein, lymphocytes, Framingham scores and predicted coronary risks. The odds ratio (OR) of metabolic syndrome for steatohepatitis was the highest (OR = 9.9), followed by high glucose status (OR = 4.5) and obesity (OR = 3.6). The highest area under the ROC curve was metabolic syndrome (area = 0.80), followed by obesity (0.75) and high glucose level (0.73). Metabolic syndrome was the highest population-attributable risk factor (0.59). Significant interaction was found with a three-factor model, including obesity, metabolic syndrome and Framingham risk status, with lesser average prediction error (22.6%), higher average cross-validation consistency (6.3) and lower average prediction error (24.3%). Compared with persons with no risk factors, OR increased as the number of risk factors increased (OR = 3.0 with one risk factor, 17.5 with two risk factors, 10.8 with three risk factors, respectively). CONCLUSION: Metabolic syndrome, inflammation markers and atherosclerotic risk scores are significantly related to steatohepatitis status among the healthy examinee population in Taiwan. PMID- 16707023 TI - Population-based laboratory surveillance of Hafnia alvei isolates in a large Canadian health region. AB - BACKGROUND: Hospital-based series have characterized Hafnia alvei primarily as an infrequent agent of polymicrobial nosocomial infections in males with underlying illness. METHODS: We conducted population-based laboratory surveillance in the Calgary Health Region during 2000-2005 to define the incidence, demographic risk factors for acquisition, and anti-microbial susceptibilities of Hafnia alvei isolates. RESULTS: A total of 138 patients with Hafnia alvei isolates were identified (2.1/100,000/year) and two-thirds were of community onset. Older age and female gender were important risk factors for acquisition. The most common focus of isolation was urine in 112 (81%), followed by lower respiratory tract in 10 (7%), and soft tissue in 5 (4%), and the majority (94; 68%) were mono microbial. Most isolates were resistant to ampicillin (111;80%), cephalothin (106; 77%), amoxicillin/clavulanate (98; 71%), and cefazolin (95; 69%) but none to imipenem or ciprofloxacin. CONCLUSION: Hafnia alvei was most commonly isolated as a mono-microbial etiology from the urinary tract in women from the community. This study highlights the importance of population-based studies in accurately defining the epidemiology of an infectious disease. PMID- 16707024 TI - Antithrombin III (AT) and recombinant tissue plasminogen activator (R-TPA) used singly and in combination versus supportive care for treatment of endotoxin induced disseminated intravascular coagulation (DIC) in the neonatal pig. AB - BACKGROUND: Disseminated intravascular coagulation (DIC) is a pathological disturbance of the complex balance between coagulation and anticoagulation that is precipitated by vascular injury, acidosis, endotoxin release and/or sepsis and characterized by severe bleeding and excessive clotting. The innately low levels of coagulation factors found in newborn infants place them at extremely high risk for DIC. Anecdotal reports suggest that either anticoagulant or fibrinolytic therapy may alleviate some of the manifestations of DIC. To test the hypothesis that replacement of both anticoagulants and fibrinolytics may improve survival and outcome better than either single agent or supportive care alone, we utilized a neonatal piglet model of endotoxin-induced DIC. METHODS: DIC was induced in twenty-seven neonatal pigs (7 to 14 days of age) by intravenous administration of E. coli endotoxin (800 microg/kg over 30 min). The piglets were divided into 4 groups on the basis of treatment protocol [A: supportive care alone; B: Antithrombin III (AT, 50 microg/kg bolus, 25 microg/kg per hr continuous infusion) and supportive care; C: Recombinant Tissue Plasminogen Activator (R TPA, 25 microg/kg per hr continuous infusion) and supportive care; D: AT, R-TPA and supportive care] and monitored for 3 primary outcome parameters (survival time, macroscopic and microscopic organ involvement) and 4 secondary outcome parameters (hematocrit; platelet count; fibrinogen level; and antithrombin III level). RESULTS: Compared with supportive care alone, combination therapy with AT and R-TPA resulted in a significant improvement of survival time, hematocrit, AT level, macroscopic and microscopic organ involvement, p < 0.05. Compared with supportive care alone, R-TPA alone significantly reduced macroscopic organ involvement and AT alone increased AT levels. CONCLUSION: The findings suggest that combining AT, R-TPA and supportive care may prove more advantageous in treating the clinical manifestations of DIC in this neonatal pig model than either single modality or supportive care alone. PMID- 16707021 TI - Downregulation of survivin expression and concomitant induction of apoptosis by celecoxib and its non-cyclooxygenase-2-inhibitory analog, dimethyl-celecoxib (DMC), in tumor cells in vitro and in vivo. AB - BACKGROUND: 2,5-Dimethyl-celecoxib (DMC) is a close structural analog of the selective cyclooxygenase-2 (COX-2) inhibitor celecoxib (Celebrex) that lacks COX 2-inhibitory function. However, despite its inability to block COX-2 activity, DMC is able to potently mimic the anti-tumor effects of celecoxib in vitro and in vivo, indicating that both of these drugs are able to involve targets other than COX-2 to exert their recognized cytotoxic effects. However, the molecular components that are involved in mediating these drugs' apoptosis-stimulatory consequences are incompletely understood. RESULTS: We present evidence that celecoxib and DMC are able to down-regulate the expression of survivin, an anti apoptotic protein that is highly expressed in tumor cells and known to confer resistance of such cells to anti-cancer treatments. Suppression of survivin is specific to these two drugs, as other coxibs (valdecoxib, rofecoxib) or traditional NSAIDs (flurbiprofen, indomethacin, sulindac) do not affect survivin expression at similar concentrations. The extent of survivin down-regulation by celecoxib and DMC in different tumor cell lines is somewhat variable, but closely correlates with the degree of drug-induced growth inhibition and apoptosis. When combined with irinotecan, a widely used anticancer drug, celecoxib and DMC greatly enhance the cytotoxic effects of this drug, in keeping with a model that suppression of survivin may be beneficial to sensitize cancer cells to chemotherapy. Remarkably, these effects are not restricted to in vitro conditions, but also take place in tumors from drug-treated animals, where both drugs similarly repress survivin, induce apoptosis, and inhibit tumor growth in vivo. CONCLUSION: In consideration of survivin's recognized role as a custodian of tumor cell survival, our results suggest that celecoxib and DMC might exert their cytotoxic anti-tumor effects at least in part via the down-regulation of survivin - in a manner that does not require the inhibition of cyclooxygenase-2. Because inhibition of COX-2 appears to be negligible, it might be worthwhile to further evaluate DMC's potential as a non-coxib alternative to celecoxib for anti cancer purposes. PMID- 16707025 TI - Dataset of manually measured QT intervals in the electrocardiogram. AB - BACKGROUND: The QT interval and the QT dispersion are currently a subject of considerable interest. Cardiac repolarization delay is known to favor the development of arrhythmias. The QT dispersion, defined as the difference between the longest and the shortest QT intervals or as the standard deviation of the QT duration in the 12-lead ECG is assumed to be reliable predictor of cardiovascular mortality. The seventh annual PhysioNet/Computers in Cardiology Challenge, 2006 addresses a question of high clinical interest: Can the QT interval be measured by fully automated methods with accuracy acceptable for clinical evaluations? METHOD: The PTB Diagnostic ECG Database was given to 4 cardiologists and 1 biomedical engineer for manual marking of QRS onsets and T-wave ends in 458 recordings. Each recording consisted of one selected beat in lead II, chosen visually to have minimum baseline shift, noise, and artifact.In cases where no T wave could be observed or its amplitude was very small, the referees were instructed to mark a 'group-T-wave end' taking into consideration leads with better manifested T wave.A modified Delphi approach was used, which included up to three rounds of measurements to obtain results closer to the median. RESULTS: A total amount of 2*5*548 Q-onsets and T-wave ends were manually marked during round 1. To obtain closer to the median results, 8.58 % of Q-onsets and 3.21 % of the T-wave ends had to be reviewed during round 2, and 1.50 % Q-onsets and 1.17 % T-wave ends in round 3. The mean and standard deviation of the differences between the values of the referees and the median after round 3 were 2.43 +/- 0.96 ms for the Q-onset, and 7.43 +/- 3.44 ms for the T-wave end. CONCLUSION: A fully accessible, on the Internet, dataset of manually measured Q-onsets and T wave ends was created and presented in additional file: 1 (Table 4) with this article. Thus, an available standard can be used for the development of automated methods for the detection of Q-onsets, T-wave ends and for QT interval measurements. PMID- 16707027 TI - Pre-morbid IQ and response to routine outcome assessment. AB - BACKGROUND: Routine use of standardized outcome measures within adult mental health services is compulsory in many countries, but not common in clinical practice. As well as political and professional factors, one reason may be that there is little empirical evidence of benefit for patients. It is therefore important to identify predictors of response. Pre-morbid intellectual functioning has predicted response to other interventions. This paper tests the hypothesis that pre-morbid IQ impacts on the effectiveness of routine assessment and feedback of outcome measures. METHOD: The Feedback of Outcomes to Users and Staff (FOCUS) single-blind randomized controlled trial (RCT) investigated routine completion and feedback of outcome measures for 160 (101 intervention, 59 control) adult mental health service patients in Croydon, South London (International Standard RCT Number 16971059). RESULTS: Patients in the top quarter (pre-morbid IQ>110) differentially improved in patient-rated unmet need (adjusted difference 3.4, 95% CI 0.8 to 5.9, p=0.012) and in the top half (pre morbid IQ>99) in quality of life (adjusted difference -0.6, 95% CI -1.1 to -0.1, p=0.02). The top quarter result remained when controlling for the influence of baseline (p=0.004) and baseline plus follow-up variables (p=0.047). CONCLUSIONS: Feedback of routine outcome measures may improve outcome for patients with higher pre-morbid IQ. There is a need to understand more how routine outcome information is used by staff and patients to inform care. If the goal of routine use of outcome measures is to benefit patients directly, then targeted policies are required. PMID- 16707026 TI - Evidence from bioinformatics, expression and inhibition studies of phosphoinositide-3 kinase signalling in Giardia intestinalis. AB - BACKGROUND: Giardia intestinalis is a parasitic protozoan and major cause of diarrhoeal disease. Disease transmission is dependent on the ability of the parasite to differentiate back and forth between an intestine-colonising trophozoite and an environmentally-resistant infective cyst. Our current understanding of the intracellular signalling mechanisms that regulate parasite replication and differentiation is limited, yet such information could suggest new methods of disease control. Phosphoinositide-3 kinase (PI3K) signalling pathways have a central involvement in many vital eukaryotic processes, such as regulation of cell growth, intracellular membrane trafficking and cell motility. Here we present evidence for the existence of functional PI3K intracellular signalling pathways in G. intestinalis. RESULTS: We have identified and characterised two genes, Gipi3k1 and Gipi3k2, which encode putative PI3Ks. Both genes are expressed in trophozoites and encysting cells, suggesting a possible role of GiPI3K1 and GiPI3K2 in regulating giardial growth and differentiation. Extensive nucleotide and amino acid sequence characterisation predicts that both encoded PI3Ks are functional as indicated by the presence of highly conserved structural domains and essential catalytic residues. The inhibitory effect of the PI3K inhibitor LY294002 on trophozoite proliferation also supports their functionality. Phylogenetic analysis supports the identity of GiPI3K1 as a Class I isoform and GiPI3K2 as a Class III isoform. In addition, giardial genes encoding putative homologues of phosphoinositide-metabolising enzymes such as PTEN, MTM, PIPkin and PI 5-phosphatase as well as downstream effectors with phosphoinositide-binding domains have been identified, placing GiPI3K1 and GiPI3K2 in a broader signalling context. Compared with twenty-six PI3Ks from other organisms, GiPI3K1 and GiPI3K2 are unique in that they contain large insertions within their highly conserved kinase domains. The function of these insertions is unknown; however we show here that they are not intron-derived and would probably not hinder substrate binding. These insertions may represent a plausible drug target. CONCLUSION: G. intestinalis encodes and expresses two putative PI3Ks, at least one of which appears to be required during normal parasite proliferation. The identification of Class I and Class III but not Class II isoforms suggests that both extracellularly-initiated signalling (Class I regulated) and intracellular vesicle trafficking (Class III-regulated) might be controlled by PI3Ks in G. intestinalis. The presence of genes encoding putative homologues of phosphoinositide-metabolising enzymes and downstream effectors in the G. intestinalis genome further suggests that the overall architecture of PI3K signalling may be comparable with pathways present in other better-studied organisms. PMID- 16707028 TI - The alliance against depression: 2-year evaluation of a community-based intervention to reduce suicidality. AB - BACKGROUND: The global burden and large diagnostic and therapeutic deficits associated with depressive disorders call for intervention programs. The aim of the Nuremberg Alliance against Depression (NAD) is to establish and to assess the effectiveness of a four-level intervention program for improving the care of patients with depression. METHOD: A 2-year intervention program was performed in Nuremberg (480000 inhabitants) at four levels: training of family doctors and support through different methods; a public relations campaign informing about depression; cooperation with community facilitators (teachers, priests, local media, etc.); and support for self-help activities as well as for high-risk groups. The effects of the 2-year intervention on the number of suicidal acts (completed suicides plus suicide attempts, main outcome criterion) were evaluated with respect to a 1-year baseline and a control region (Wuerzburg, 270,000 inhabitants). RESULTS: Compared to the control region, a reduction in frequency of suicidal acts was observed in Nuremberg during the 2-year intervention (2001 v. 2000: -19.4%; p< or =0.082; 2002 v. 2000: -24%, p< or =0.004). Considering suicide attempts only (secondary outcome criterion), the same effect was found (2001 v. 2000: -18.3%, p< or =0.023; 2002 v. 2000: -26.5%, p<0.001). The reduction was most noticeable for high-risk methods (e.g. hanging, jumping, shooting). Concerning completed suicides, there were no significant differences compared to the control region. CONCLUSIONS: The NAD appeared to be effective in reducing suicidality. It provides a concept as well as many methods that are currently being implemented in several other intervention regions in Germany and in other countries. PMID- 16707029 TI - A pilot study of positive mood induction in euthymic bipolar subjects compared with healthy controls. AB - BACKGROUND: Demonstrating differences between euthymic bipolar subjects and healthy controls in response to positive (happy) mood induction may help elucidate how mania evolves. This pilot study evaluates the Go task in a reward paradigm as a method for inducing a happy mood state and compares the response of euthymic bipolar subjects and healthy controls. METHOD: The Sense of Hyperpositive Self Scale, the Tellegen positive and negative adjectives, the Global-Local task and a visual analogue scale for measuring positive affect were administered to 15 euthymic bipolar subjects and 19 age-and-sex-matched healthy control subjects before and after they had performed the Go task in a reward paradigm. RESULTS: Significant differences were found between subjects and controls on several measures at each time-point but there were no differences across the groups across time except for the visual analogue scales, where subjects had a more sustained duration in self-reported happiness compared with controls. CONCLUSIONS: This pilot study has shown that a positive affect can be induced in bipolar subjects and controls which can be demonstrated by changes in scores on several tasks. However, only the visual analogue scales showed a significant difference between cases and controls over time. Such tests may prove valuable in furthering understanding about the evolution of manic mood states. PMID- 16707030 TI - Potential biases in estimates of hepatitis C RNA clearance in newly acquired hepatitis C infection among a cohort of injecting drug users. AB - Estimates of hepatitis C virus (HCV) clearance following acute infection range from 14 to 46%. This wide range is likely to be due to the characteristics of the populations studied and analysis methods. This paper examines how differing definitions of clearance parameters affect estimates of viral clearance in a cohort of 85 injecting drug users with newly acquired HCV infection. Kaplan-Meier estimates of time to HCV clearance were determined using varying definitions of eligible cohort, viral clearance, date of infection and date of clearance. Based on which combinations of definitions were used, the number of subjects eligible for analysis ranged from 27 to 75, clearance rate ranged from 14 to 68% and time to achieving 25% clearance ranged from approximately 5 months to 14 months. Standardized definitions and methodologies are required to enable valid comparisons of rates of clearance across newly acquired HCV infection natural history studies. PMID- 16707031 TI - Social mixing patterns for transmission models of close contact infections: exploring self-evaluation and diary-based data collection through a web-based interface. AB - Although mixing patterns are crucial in dynamic transmission models of close contact infections, they are largely estimated by intuition. Using a convenience sample (n=73), we tested self-evaluation and prospective diary surveys with a web based interface, in order to obtain social contact data. The number of recorded contacts was significantly (P<0.01) greater on workdays (18.1) vs. weekend days (12.3) for conversations, and vice versa for touching (5.4 and 7.2 respectively). Mixing was highly assortative with age for both (adults contacting other adults vs. 0- to 5-year-olds, odds ratio 8.9-10.8). Respondents shared a closed environment significantly more often with >20 other adults than with >20 children. The difference in number of contacts per day was non-significant between self-evaluation and diary (P=0.619 for conversations, P=0.125 for touching). We conclude that self-evaluation could yield similar results to diary surveys for general or very recent mixing information. More detailed data could be collected by diary, at little effort to respondents. PMID- 16707032 TI - Enhanced efficacy of both typical and atypical antipsychotic drugs by adjunctive alpha2 adrenoceptor blockade: experimental evidence. AB - Adjunctive treatment with the selective alpha2 adrenoceptor antagonist idazoxan augments the effect of conventional antipsychotics in treatment-resistant schizophrenics comparing favourably with clozapine. Clozapine has high affinity for alpha2 adrenoceptors. Previously, we found that adjunctive idazoxan treatment to the dopamine (DA) D2/3 antagonist raclopride enhanced raclopride-induced effects in an animal model of antipsychotic activity (conditioned avoidance response, CAR) and, similarly to clozapine, reversed the disruption of working memory induced by N-methyl-D-aspartate receptor blockade in rats with a concomitant increase in prefrontal DA efflux. To further investigate the significance of alpha2 adrenoceptor affinity for antipsychotic efficacy, we here investigated, in rats, the effects of adjunctive idazoxan treatment to low doses of a typical (haloperidol) and an atypical (olanzapine) antipsychotic drug, both lacking appreciable alpha2 adrenoceptor affinity, on (i) CAR; (ii) catalepsy; and (iii) DA output in the prefrontal cortex and the nucleus accumbens using microdialysis. Adjunctive treatment with idazoxan to haloperidol or olanzapine enhanced suppression of CAR to a level predicting sufficient antipsychotic activity, increased DA output preferentially in the prefrontal cortex, and reversed haloperidol-induced catalepsy. Our data confirm and extend our previous findings as well as clinical observations, and suggest that adjunctive alpha2 adrenoceptor blockade both typical and atypical antipsychotic drugs, lacking appreciable affinity for the alpha2 adrenoceptor, may contribute to a more advantageous therapeutical profile of these drugs in schizophrenia treatment, allowing for reduced DA D2 occupancy and reduction of unwanted side-effects. PMID- 16707033 TI - Cholesterol content in brains of suicide completers. AB - An association between low levels of serum cholesterol and violent or suicidal behaviour has frequently been reported, but it remains unclear how cholesterol in the peripheral system might be related to the brain functions involved in mediating suicidal behaviour. To our knowledge, there have been no previous studies aimed at answering the important question of whether there are differences in cholesterol within the brains of suicide completers. In the present study, cholesterol content was measured in cortical and subcortical tissue of brains from 41 male suicide completers and 21 male controls that died of sudden causes with no direct influence on brain tissue. No significant differences in cholesterol content were found between suicides and controls in the frontal cortex, amygdala or hippocampus. However, when the suicide completers were stratified into violent (n=31) or non-violent (n=10) groups based on the method of death, violent suicides were found to have lower grey-matter cholesterol content overall compared to non-violent suicides [F(1,111)=4.75, p=0.03], specifically in the frontal cortex (t=-4.16, d.f.=37, p<0.0005). Further exploration of the frontal cortex revealed that violent suicides had lower cholesterol content compared to non-violent suicides in the orbitofrontal cortex (t=-2.01, d.f.=36, p=0.05) and the ventral prefrontal cortex (t=-2.49, d.f.=37, p=0.02). This study represents the first direct examination of cholesterol content in brain tissue from suicide completers, and the present findings provide added support for the relationship between low cholesterol and violent or suicidal behaviour. PMID- 16707034 TI - Avoiding bias in parasite excretion estimates: the effect of sampling time and type of faeces. AB - The study of host-parasite relationships usually requires reliable estimates of parasite intensity, which is often estimated from parasite propagule concentration in faeces. However, parasite excretion in faeces may be subject to variation due to endogenous or exogenous factors that must be identified to obtain reliable results. We analysed the effect of the hour of sample collection on propagule counts of 2 intestinal parasites infecting the red-legged partridge: the capillarid nematode Aonchoteca caudinflata and coccidia of the genus Eimeria (Protozoa). Also, we test whether there are differences in propagule counts between caecal and intestinal faeces. Individual faecal samples from infected birds were collected daily at 4 different hours during several days. The hour of the day exerted a very strong effect on propagule counts, excretion of both types of parasites showing a clear and constant increase from dawn to dusk. Also, capillarid eggs were more abundant in intestinal than in caecal faeces, whereas the inverse pattern was found for coccidian oocysts. Standardization of the hour of sample collection or statistical control of this variable is recommendable to prevent bias. Similarly, in bird species with long caeca, consistent collection of one type of faeces may avoid significant errors in parasite burden estimates. PMID- 16707035 TI - Why are head and neck cancer clinicians not measuring quality of life? AB - AIM: To quantify and qualify the use of quality of life (QOL) measures by head and neck cancer clinicians and to identify any impediments to the use of these measures. METHODS: Questionnaire survey of members of the Australia and New Zealand Head and Neck Society. RESULTS: One hundred and twenty-eight of 187 (68.5 per cent) responded. Only 43 (34 per cent) had ever used a QOL questionnaire (QLQ), and only 17 (13 per cent) were currently using one. Impediments to QLQ use included clinicians' perceptions that QLQs were too time-consuming and conferred no proven benefit for clinical management. Nevertheless, 113 (88 per cent) respondents indicated willingness to use a minimum core QLQ--for routine clinical use and for research--but indicated a preference for a short (10-15 questions), quick (less than 10 minutes) questionnaire. CONCLUSIONS: Most head and neck cancer clinicians did not use a QOL measure routinely, with impediments to routine use being mainly clinician-based. Most respondents would use a minimum core QOL measure, especially if it were a short, quick consensus questionnaire. PMID- 16707036 TI - Unusual otological manifestations in Camurati-Engelmann's disease. AB - Camurati-Engelmann's disease (CED) is a rare hereditary disorder affecting mainly the diaphysis of long bones but multiple cranial nerve deficits may also develop secondary to bony sclerosis of their foramina, including visual loss, facial palsy, deafness, vestibular disturbances and sensory deficits along the distribution of the trigeminal nerve. Deafness has been reported in about 18 per cent of these cases due to narrowing of the internal auditory canals caused by bony encroachment on nerves and vessels. We report an extremely rare case of a patient with CED who presented with deafness due to gross abnormalities affecting both middle ear and cochlea. The issues relating to the management of these patients with temporal bone involvement are discussed. PMID- 16707037 TI - Surgical management of trigeminal neuromas: a report of eight cases. AB - OBJECTIVE: Trigeminal neuromas are rare tumours that may involve any part of the Vth nerve complex, including extracranial peripheral divisions of the nerve. A series of eight patients with intracranial trigeminal neuromas who underwent surgical management are presented. METHODS: The hospital records and radiological images were reviewed with regard to clinical presentation, surgical approach, operative findings, peri-operative morbidity and neurological outcome, and percentage of tumour recurrence. RESULTS: The principal presenting symptoms were those of involvement of the trigeminal nerve with sensory impairment in one or more of the three divisions. Tumour location was the prime determinant of surgical approach. Lateral skull base approaches were used as they are considered to be superior for identifying tumour origin, extension, and relationship to surrounding structures. Total excision of the tumour was carried out in three of the eight patients. In the remaining five patients some tumour was left purposely in order to minimize neurological deficit and optimize post-operative quality of life. There was no peri-operative mortality or major morbidity in our series. Five patients experienced symptomatic tumour recurrence and revision surgery was performed. CONCLUSION: Management of trigeminal neuromas is complex and requires a multidisciplinary approach. Pre-operative surgical planning allows tumour removal with preservation of important neural structures in the majority of cases. For large tumours occupying both the middle and posterior cranial fossae, the retrosigmoid/retrolabyrinthine/middle cranial fossa approach provides good exposure and results in minimal brain retraction. A Fisch type C approach is necessary for the largest tumours. Long-term follow up with interval imaging is mandatory to exclude long-term tumour recurrence. PMID- 16707038 TI - Intratympanic gentamicin for Meniere's disease: effect on quality of life as assessed by Glasgow benefit inventory. AB - OBJECTIVES: To evaluate patients' quality of life after receiving intratympanic gentamicin as treatment for Meniere's disease. DESIGN: Retrospective study using the Glasgow benefit inventory scale (GBI) questionnaire. PARTICIPANTS: Patients diagnosed with Meniere's disease, according to the American Academy of Otolaryngology-Head and Neck Surgery (AAO-HNS) diagnostic criteria (refractory to medical management), who were treated with intratympanic gentamicin injection. MATERIALS AND METHODS: This retrospective study included all patients diagnosed with Meniere's disease (according to AAO-HNS criteria; refractory to medical management) undergoing gentamicin labyrinthectomy in our unit over the preceding three years. Other causes of hearing loss were ruled out as all patients underwent magnetic resonance imaging, electronystagmography and sway magnetometry. Patients underwent gentamicin infiltration at a concentration of 2 ml of 30 mg/ml and were reviewed six weeks later and received a repeat injection if no benefit was evident. Six months after their last follow up, they were sent a GBI questionnaire. Twenty-one questionnaires were posted, and the response rate was 81 per cent. RESULTS: As per the responses received, the total benefit of intratympanic gentamicin injection was found to be +30.3. The three components of GBI were analysed separately; it was found that general benefit was greatest (+33.3), followed by physical benefit (+28.1) and social benefit (+21.6). CONCLUSION: This study suggests that gentamicin definitely improves quality of life in patients with Meniere's disease and should be the first line of treatment if medical management fails. PMID- 16707039 TI - Period and cohort dynamics in fertility norms at the onset of the demographic transition in Kenya 1978-1998. AB - A characteristic of African pre-transitional fertility regimes is large ideal family size. This has been used to support claims of cultural entrenchment of high fertility. Yet in Kenya fertility rates have fallen. In this paper this fall is explored in relation to trends in fertility norms and attitudes using four sequential cross-sectional surveys spanning the fertility transition in Kenya (1978, 1984, 1989 and 1998). The most rapid fall in the reported ideal family size occurred between 1984 and 1989, whilst the most rapid fall in the total fertility rate occurred 5 to 10 years later, between 1989 and 1998. Thus these data, spanning the fertility transition in Kenya, support the traditional demographic model that demand for fertility limitation drives fertility decline. These data also suggest that the decline in fertility norms over time was partly a period effect, as the reported ideal family size was seen to fall simultaneously in all age cohorts, and partly a cohort effect, as older age cohorts reporting higher ideal family sizes were replaced by younger cohorts reporting lower ideal family sizes. These data also suggest that a new fertility norm of four children may have developed by 1989 and continued until 1998. This is consistent with, and perhaps could have been used to predict, the stall in the Kenyan fertility decline after 1998. PMID- 16707040 TI - Disability life expectancy for the elderly, city of Sao Paulo, Brazil, 2000: gender and educational differences. AB - There is evidence that 'health life expectancy' (expected number of years to be lived in health) differs by socioeconomic status. Time spent in health or disability plays a critical role in the use of health care services. The objective of this study was to estimate 'disability life expectancy' by age, gender and education attainment for the elderly of the city of Sao Paulo, Brazil, in the year 2000. Data came from the SABE database, population censuses and mortality statistics (SEADE Foundation). Life expectancy with disability was calculated using Sullivan's method on the basis of the current probability of death and prevalence of disability by educational level. The prevalence of disability increased with age, for both sexes and both levels of educational attainment studied. Men showed a lower prevalence of disability, in general, and persons with lower educational attainment showed a higher prevalence of disability. Regarding life expectancy, women could expect to live longer than men, with and without disability. For both sexes, the percentage of life expectancy lived with disability decreased with increasing educational attainment. With increasing educational attainment, the sex differences in the percentage of remaining years to be lived with disability increased for most ages. Finally, the percentage of remaining years to be lived with disability increased with age for males and females, except for males with high educational attainment between the ages 70-75 and 75-80. The results may serve as a guide for public policies in the country, since health problems faced by older persons, such as disability, are the result of a number of past experiences during their life-times, such as health care, housing conditions, hygiene practices and education. Education influences health behaviours and is related, to some extent, to all these factors. Therefore, improvements in education for the disadvantaged may improve health. PMID- 16707041 TI - Host range restriction and pathogenicity in the context of influenza pandemic. AB - Influenza A viruses cause pandemics at random intervals. Pandemics are caused by viruses that contain a hemagglutinin (HA) surface glycoprotein to which human populations are immunologically naive. Such an HA can be introduced into the human population through reassortment between human and avian virus strains or through the direct transfer of an avian influenza virus to humans. The factors that determine the interspecies transmission and pathogenicity of influenza viruses are still poorly understood; however, the HA protein plays an important role in overcoming the interspecies barrier and in virulence in avian influenza viruses. Recently, the RNA polymerase (PB2) protein has also been recognized as a critical factor in host range restriction, while the nonstructural (NS1) protein affects the initial host immune responses. We summarize current knowledge of viral factors that determine host range restriction and pathogenicity of influenza A viruses. PMID- 16707042 TI - Dengue prevention and 35 years of vector control in Singapore. AB - After a 15-year period of low incidence, dengue has reemerged in Singapore in the past decade. We identify potential causes of this resurgence. A combination of lowered herd immunity, virus transmission outside the home, an increase in the age of infection, and the adoption of a case-reactive approach to vector control contribute to the increased dengue incidence. Singapore's experience with dengue indicates that prevention efforts may not be sustainable. For renewed success, Singapore needs to return to a vector control program that is based on carefully collected entomologic and epidemiologic data. Singapore's taking on a leadership role in strengthening disease surveillance and control in Southeast Asia may also be useful in reducing virus importation. PMID- 16707043 TI - Severe community-acquired pneumonia due to Staphylococcus aureus, 2003-04 influenza season. AB - During the 2003-04 influenza season, 17 cases of Staphylococcus aureus community acquired pneumonia (CAP) were reported from 9 states; 15 (88%) were associated with methicillin-resistant S. aureus (MRSA). The median age of patients was 21 years; 5 (29%) had underlying diseases, and 4 (24%) had risk factors for MRSA. Twelve (71%) had laboratory evidence of influenza virus infection. All but 1 patient, who died on arrival, were hospitalized. Death occurred in 5 (4 with MRSA). S. aureus isolates were available from 13 (76%) patients (11 MRSA). Toxin genes were detected in all isolates; 11 (85%) had only genes for Panton-Valentine leukocidin. All isolates had community-associated pulsed-field gel electrophoresis patterns; all MRSA isolates had the staphylococcal cassette chromosome mec type IVa. In communities with a high prevalence of MRSA, empiric therapy of severe CAP during periods of high influenza activity should include consideration for MRSA. PMID- 16707044 TI - Temple monkeys and health implications of commensalism, Kathmandu, Nepal. AB - The threat of zoonotic transmission of infectious agents at monkey temples highlights the necessity of investigating the prevalence of enzootic infectious agents in these primate populations. Biological samples were collected from 39 rhesus macaques at the Swoyambhu Temple and tested by enzyme-linked immunosorbent assay, Western blot, polymerase chain reaction, or combination of these tests for evidence of infection with rhesus cytomegalovirus (RhCMV), Cercopithecine herpesvirus 1 (CHV-1), simian virus 40 (SV40), simian retrovirus (SRV), simian T cell lymphotropic virus (STLV), simian immunodeficiency virus (SIV), and simian foamy virus (SFV). Antibody seroprevalence was 94.9% to RhCMV (37/39), 89.7% to SV40 (35/39), 64.1% to CHV-1 (25/39), and 97.4% to SFV (38/39). Humans who come into contact with macaques at Swoyambhu risk exposure to enzootic primateborne viruses. We discuss implications for public health and primate management strategies that would reduce contact between humans and primates. PMID- 16707045 TI - Multidrug-resistant commensal Escherichia coli in children, Peru and Bolivia. AB - Using a rapid screening method, we investigated the prevalence of fecal carriage of antimicrobial drug-resistant Escherichia coli in 3,174 healthy children from 4 urban settings in Peru and Bolivia. High resistance rates were observed for ampicillin (95%), trimethoprim-sulfamethoxazole (94%), tetracycline (93%), streptomycin (82%), and chloramphenicol (70%). Lower resistance rates were observed for nalidixic acid (35%), kanamycin (28%), gentamicin (21%), and ciprofloxacin (18%); resistance to ceftriaxone and amikacin was uncommon (<0.5%). In a random sample of 1,080 resistant E. coli isolates, 90% exhibited a multidrug resistance (MDR) phenotype. The 2 most common MDR phenotypes (ampicillin/tetracycline/trimethoprim-sulfamethoxazole and ampicillin/tetracycline/trimethoprim-sulfamethoxazole/chloramphenicol) could be transferred en bloc in conjugation experiments. The most common acquired resistance genes were blaTEM, tet(A), tet(B), drfA8, sul1, sul2, and catI. These findings underscore the magnitude of the problem of antimicrobial drug resistance in low-resource settings and the urgent need for surveillance and control of this phenomenon. PMID- 16707046 TI - Human Streptococcus suis outbreak, Sichuan, China. AB - From mid-July to the end of August 2005, a total of 215 cases of human Streptococcus suis infections, 66 of which were laboratory confirmed, were reported in Sichuan, China. All infections occurred in backyard farmers who were directly exposed to infection during the slaughtering process of pigs that had died of unknown causes or been killed for food because they were ill. Sixty-one (28%) of the farmers had streptococcal toxic shock syndrome; 38 (62%) of them died. The other illnesses reported were sepsis (24%) and meningitis (48%) or both. All isolates tested positive for genes for tuf, species-specific 16S rRNA, cps2J, mrp, ef, and sly. A single strain of S. suis caused the outbreak, as shown by the identification of a single ribotype. The high death ratio was of concern; prohibiting backyard slaughtering ended the outbreak. PMID- 16707047 TI - Co-infections of adenovirus species in previously vaccinated patients. AB - Despite the success of the adenovirus vaccine administered to US military trainees, acute respiratory disease (ARD) surveillance still detected breakthrough infections (respiratory illnesses associated with the adenovirus serotypes specifically targeted by the vaccine). To explore the role of adenoviral co-infection (simultaneous infection by multiple pathogenic adenovirus species) in breakthrough disease, we examined specimens from patients with ARD by using 3 methods to detect multiple adenoviral species: a DNA microarray, a polymerase chain reaction (PCR)-enzyme-linked immunosorbent assay, and a multiplex PCR assay. Analysis of 52 samples (21 vaccinated, 31 unvaccinated) collected from 1996 to 2000 showed that all vaccinated samples had co-infections. Most of these co-infections were community-acquired serotypes of species B1 and E. Unvaccinated samples primarily contained only 1 species (species E) associated with adult respiratory illness. This study highlights the rarely reported phenomenon of adenoviral co-infections in a clinically relevant environment suitable for the generation of new recombinational variants. PMID- 16707048 TI - Norwalk virus-specific binding to oyster digestive tissues. AB - The primary pathogens related to shellfish-borne gastroenteritis outbreaks are noroviruses. These viruses show persistence in oysters, which suggests an active mechanism of virus concentration. We investigated whether Norwalk virus or viruslike particles bind specifically to oyster tissues after bioaccumulation or addition to tissue sections. Since noroviruses attach to carbohydrates of the histo-blood group family, tests using immunohistochemical analysis were performed to evaluate specific binding of virus or viruslike particles to oyster tissues through these ligands. Viral particles bind specifically to digestive ducts (midgut, main and secondary ducts, and tubules) by carbohydrate structures with a terminal N-acetylgalactosamine residue in an alpha linkage (same binding site used for recognition of human histo-blood group antigens). These data show that the oyster can selectively concentrate a human pathogen and that conventional depuration will not eliminate noroviruses from oyster tissue. PMID- 16707049 TI - Haemophilus influenzae type b reemergence after combination immunization. AB - An increase in Haemophilus influenzae type b (Hib) in British children has been linked to the widespread use of a diphtheria/tetanus/acellular pertussis combination vaccine (DTaP-Hib). We measured anti-polyribosyl-ribitol phosphate antibody concentration and avidity before and after a Hib booster in 176 children 2-4 years of age who had received 3 doses of DTP-Hib (either DT whole cell pertussis-Hib or DTaP-Hib) combination vaccine in infancy. We also measured pharyngeal carriage of Hib. Antibody concentrations before and avidity indices after vaccination were low (geometric mean concentration 0.46 mug/mL, 95% confidence interval [CI] 0.36-0.58; geometric mean avidity index 0.16, 95% CI 0.14-0.18) and inversely related to the number of previous doses of DTaP-Hib (p = 0.02 and p<0.001, respectively). Hib was found in 2.1% (95% CI 0.7%-6.0%) of study participants. Our data support an association between DTaP-Hib vaccine combinations and clinical Hib disease through an effect on antibody concentration and avidity. PMID- 16707050 TI - Genetic divergence of Toxoplasma gondii strains associated with ocular toxoplasmosis, Brazil. AB - Previous studies have shown a high prevalence of toxoplasmosis and the frequent occurrence of ocular disease in Brazil. To identify the genotypes of parasite strains associated with ocular disease, we compared 25 clinical and animal isolates of Toxoplasma gondii from Brazil to previously characterized clonal lineages from North America and Europe. Multilocus nested polymerase chain reaction analysis was combined with direct sequencing of a polymorphic intron to classify strains by phylogenetic methods. The genotypes of T. gondii strains isolated from Brazil were highly divergent when compared to the previously described clonal lineages. Several new predominant genotypes were identified from different regions of Brazil, including 2 small outbreaks attributable to foodborne or waterborne infection. These findings show that the genetic makeup of T. gondii is more complex than previously recognized and suggest that unique or divergent genotypes may contribute to different clinical outcomes of toxoplasmosis in different localities. PMID- 16707051 TI - Social behavior and meningococcal carriage in British teenagers. AB - Understanding predisposing factors for meningococcal carriage may identify targets for public health interventions. Before mass vaccination with meningococcal group C conjugate vaccine began in autumn 1999, we took pharyngeal swabs from approximately 14,000 UK teenagers and collected information on potential risk factors. Neisseria meningitidis was cultured from 2,319 (16.7%) of 13,919 swabs. In multivariable analysis, attendance at pubs/clubs, intimate kissing, and cigarette smoking were each independently and strongly associated with increased risk for meningococcal carriage (p<0.001). Carriage in those with none of these risk factors was 7.8%, compared to 32.8% in those with all 3. Passive smoking was also linked to higher risk for carriage, but age, sex, social deprivation, home crowding, or school characteristics had little or no effect. Social behavior, rather than age or sex, can explain the higher frequency of meningococcal carriage among teenagers. A ban on smoking in public places may reduce risk for transmission. PMID- 16707052 TI - Coccidioidomycosis as a common cause of community-acquired pneumonia. AB - The early manifestations of coccidioidomycosis (valley fever) are similar to those of other causes of community-acquired pneumonia (CAP). Without specific etiologic testing, the true frequency of valley fever may be underestimated by public health statistics. Therefore, we conducted a prospective observational study of adults with recent onset of a lower respiratory tract syndrome. Valley fever was serologically confirmed in 16 (29%) of 55 persons (95% confidence interval 16%-44%). Antimicrobial medications were used in 81% of persons with valley fever. Symptomatic differences at the time of enrollment had insufficient predictive value for valley fever to guide clinicians without specific laboratory tests. Thus, valley fever is a common cause of CAP after exposure in a disease endemic region. If CAP develops in persons who travel or reside in Coccidioides endemic regions, diagnostic evaluation should routinely include laboratory evaluation for this organism. PMID- 16707053 TI - Human rotavirus serotype G9, Sao Paulo, Brazil, 1996-2003. AB - A total of 3,101 fecal specimens were collected during an 8-year survey for rotavirus infection in Sao Paulo, Brazil. Group A rotavirus was detected in 774 (25.0%) specimens. Of these, 431 strains (55.7%) were analyzed for G and P types by reverse transcription-polymerase chain reaction; G1 was the predominant serotype (68.2%), followed by G9 (17.2%), G4 (6.3%), G2 (1.2%), G3 (0.7%), mixed infection (1.8%), and untypeable (4.6%). Both rotavirus G and P types could be established in 332 strains (77.0%). We identified the 4 most common strains worldwide: P[8]G1 (66.6%), P[4]G2 (1.0%), P[8]G3 (0.6%), and P[8]G4 (7.2%). Among the single G9 strains detected, VP4 genotyping showed that P[8]G9 was the most prevalent, followed by P[4]G9 and P[6]G9. The emergence and high frequency of rotavirus G9 in Sao Paulo, Brazil, and other parts of the world will affect the development and evaluation of future vaccines. PMID- 16707055 TI - 2,500-year evolution of the term epidemic. AB - The term epidemic (from the Greek epi [on] plus demos [people]), first used by Homer, took its medical meaning when Hippocrates used it as the title of one of his famous treatises. At that time, epidemic was the name given to a collection of clinical syndromes, such as coughs or diarrheas, occurring and propagating in a given period at a given location. Over centuries, the form and meaning of the term have changed. Successive epidemics of plague in the Middle Ages contributed to the definition of an epidemic as the propagation of a single, well-defined disease. The meaning of the term continued to evolve in the 19th-century era of microbiology. Its most recent semantic evolution dates from the last quarter of the 20th century, and this evolution is likely to continue in the future. PMID- 16707054 TI - Human parechovirus types 1, 2 and 3 infections in Canada. AB - A new reverse transcription-polymerase chain reaction assay was developed for identification of 28 Canadian human parechovirus (HPeV) isolates, including 20 HPeV-1, 3 HPeV-2, and 5 HPeV-3, recovered from 1985 to 2004. All HPeV-1 isolates but 1 were genetically distinct from the Harris reference strain. One HPeV-2 isolate was related to the Williamson strain; the other 2 were related to the Connecticut strain. HPeV-3 isolates clustered together. Seventy-five percent of isolates were recovered during the typical enterovirus season. All patients but 1 were children with a mean age of 14.6 months, 6.3 months, and 0.7 months for HPeV 1, HPeV-2, and HPeV-3 patients, respectively. All HPeV-2- and HPeV-3-infected children were hospitalized with a diagnosis of viremia or sepsis. HPeV-1- infected children had bronchiolitis diagnosed in 50% of the cases, with few cases of pneumonia and enteritis. Two infected patients (1 child with leukemia and a 78 year-old woman) died of septic shock and severe pneumonia, respectively. PMID- 16707056 TI - VIM-1 metallo-beta-lactamase in Acinetobacter baumannii. AB - In 2004 and 2005, 5 metallo-beta-lactamase (MBL)-positive Acinetobacter baumannii isolates were found in 2 Greek hospitals. Isolates were unrelated and carried blaVIM-1 in a class 1 integron; bla(OXA-51-) and bla(OXA-58-like) carbapenemase genes were also detected. VIM-1 MBL in Acinetobacter spp. causes concern, given the increasing resistance of this species. PMID- 16707057 TI - Acanthamoeba encephalitis in patient with systemic lupus, India. AB - We report a fatal case of encephalitis caused by Acanthamoeba in a 24-year-old woman from India with systemic lupus erythematosus. Diagnosis was made by identification of amebas in brain sections by immunofluorescence analysis and confirmed by demonstrating Acanthamoeba mitochondrial 16S rRNA gene DNA in brain tissue sections. PMID- 16707058 TI - Bordetella pertussis, Finland and France. AB - We used pulsed-field gel electrophoresis analysis and genotyping to compare clinical isolates of Bordetella pertussis recovered since the early 1990s in Finland and France, 2 countries with similar histories of long-term mass vaccination with whole-cell pertussis vaccines. Isolates from both countries were similar genetically but varied temporally. PMID- 16707059 TI - Guillain-Barre syndrome, greater Paris area. AB - We studied 263 cases of Guillain-Barre syndrome from 1996 to 2001, 40% of which were associated with a known causative agent, mainly Campylobacter jejuni (22%) or cytomegalovirus (15%). The cases with no known agent (60%) peaked in winter, and half were preceded by respiratory infection, influenza-like syndrome, or gastrointestinal illness. PMID- 16707060 TI - Francisella tularensis in rodents, China. AB - A total of 420 rodents in China were examined for Francisella tularensis by polymerase chain reaction. The infection rates were 4.76% in total, and 11.65%, 10.00%, 6.56%, 1.77%, and 0% in Jilin, Xinjiang, Heilongjiang, Inner Mongolia, and Zhejiang, respectively. Sequence analysis showed that all the detected agents belonged to F. tularensis subsp. holarctica. PMID- 16707061 TI - Preventing zoonotic influenza virus infection. AB - We evaluated 49 swine industry workers and 79 nonexposed controls for antibodies to swine influenza viruses. Multivariate modeling showed that workers who seldom used gloves (odds ratio [OR] 30.3) or who smoked (OR 18.7) most frequently had evidence of previous H1N1 swine virus. These findings may be valuable in planning for pandemic influenza. PMID- 16707063 TI - Cowpox virus transmission from rats to monkeys, the Netherlands. AB - We report an outbreak of cowpox virus among monkeys at a sanctuary for exotic animals. Serologic analysis and polymerase chain reaction were performed on blood and swab samples from different rodent species trapped at the sanctuary during the outbreak. Sequence comparison and serologic results showed that brown rats (Rattus norvegicus) transmitted the virus to monkeys. PMID- 16707062 TI - Antiretroviral drug resistance and routine therapy, Cameroon. AB - Among 128 patients routinely receiving highly active antiretroviral therapy in an HIV/AIDS outpatient clinic in Cameroon, 16.4% had drug resistance after a median of 10 months. Of these, 12.5% had resistance to nucleoside reverse transcriptase inhibitors (NRTIs), 10.2% to non-NRTIs, and 2.3% to protease inhibitors. PMID- 16707064 TI - Pasture types and Echinococcus multilocularis, Tibetan communities. AB - Our study showed that open pastures had more small mammal burrows than fenced pastures in Tibetan pastoralist communities in 2003. This characteristic was linked to a higher prevalence of Echinococcus multilocularis in dogs and indicates that pasture type may affect E. multilocularis transmission. PMID- 16707065 TI - Class 1 integrons in resistant Escherichia coli and Klebsiella spp., US hospitals. AB - We examined Escherichia coli and Klebsiella spp. from US hospitals for class 1 integrons. Of 320 isolates, 181 (57%) were positive; association of integrons with resistance varied by drug and organism. Thus, determining integron epidemiology will improve understanding of how antibacterial resistance determinants spread in the United States. PMID- 16707066 TI - Hantaviruses in Serbia and Montenegro. AB - Hantaviruses are endemic in the Balkan Peninsula. An outbreak of hemorrhagic fever with renal syndrome occurred in 2002 in Serbia and Montenegro. The epidemiologic characteristics and genetic relatedness of Dobrava/Belgrade virus strains responsible for most cases are described. PMID- 16707067 TI - Raccoons and skunks as sentinels for enzootic tularemia. AB - We analyzed sera from diverse mammals of Martha's Vineyard, Massachusetts, for evidence of Francisella tularensis exposure. Skunks and raccoons were frequently seroreactive, whereas white-footed mice, cottontail rabbits, deer, rats, and dogs were not. Tularemia surveillance may be facilitated by focusing on skunks and raccoons. PMID- 16707068 TI - West Nile Virus infection in humans and horses, Cuba. AB - A surveillance system to detect West Nile virus (WNV) was established in Cuba in 2002. WNV infection was confirmed by serologic assays in 4 asymptomatic horses and 3 humans with encephalitis in 2003 and 2004. These results are the first reported evidence of WNV activity in Cuba. PMID- 16707069 TI - Mixed Cryptosporidium infections and HIV. AB - Mixed Cryptosporidium infections were detected in 7 of 21 patients with a diagnosis of rare Cryptosporidium canis or C. felis infections; 6 patients were infected with 2 Cryptosporidium spp. and 1 patient with 3 species. Mixed infections may occur more frequently than previously believed and should be considered when assessing cryptosporidiosis. PMID- 16707070 TI - Randomised clinical trial, observational study and assessment of cost effectiveness of the treatment of varicose veins (REACTIV trial). AB - OBJECTIVES: To establish the cost-effectiveness of surgery and sclerotherapy for the treatment of varicose veins. DESIGN: Randomised controlled trials (RCTs) were carried out for conservative treatment, sclerotherapy and surgery for varicose veins. An economic analysis was carried out alongside the randomised trial. Economic modelling was undertaken based on the primary data collection and a literature review (database searches undertaken in April 2000 and updated in March 2001). SETTING: Primary data collection was from a large district general hospital and a teaching hospital both in England over a 2-year period from January 1999. Cost-effectiveness analysis and economic modelling were carried out using an NHS perspective. PARTICIPANTS: A total of 1009 patients were recruited. INTERVENTIONS: Thirty-four patents were randomised in Group 1 (minor varicose veins with no reflux, randomised between conservative treatment and sclerotherapy), 77 in Group 2 (moderate varicose veins with reflux, randomised between surgery and sclerotherapy) and 246 in Group 3 (severe varicose veins with reflux, randomised between conservative treatment and surgery). The remaining 652 patients formed the observational part of the study. MAIN OUTCOME MEASURES: The cost-effectiveness analysis was based on NHS treatment costs for the 2002--3 financial year, and utilities based on the Short Form 6D (SF-6D) preference-based health measure. For the clinical trial, the outcome measures were health-related quality of life (HRQoL) [Short Form with 36 Items (SF-36), EuroQol quality of life questionnaire (EQ-5D), visual analogue scale (VAS) and standard gamble], symptomatic relief, anatomical extent (for which a new classification was developed and validated), patient satisfaction and the incidence of complications. RESULTS: Of the RCTs, only the Group 3 trial was large enough to provide clear results. This showed that surgical treatment produced better results than conservative treatment in terms of HRQoL, symptomatic relief, anatomical extent and patient satisfaction. Clinical outcomes of surgery and sclerotherapy showed significant improvement in the extent of varicose veins, symptomatic and HRQoL parameters. Cost-effectiveness analysis based on the Group 3 trial showed that the surgery produced an estimated discounted benefit of 0.054 quality-adjusted life-year (QALY) over a 2-year period, with an additional discounted cost of pound 387.45, giving an incremental cost-effectiveness ratio (ICER) of pound 7175 per QALY. Economic modelling suggested that surgery produced a still greater benefit when considered with a 10-year time horizon, with an ICER of pound 1936 per QALY. Injection sclerotherapy produced an incremental benefit of approximately 0.044 QALY at a cost of pound 155 when compared with conservative treatment, giving an ICER of pound 3500 per QALY. When surgery was compared with sclerotherapy, surgery produced greater benefit with a lower ICER (showing extended dominance). CONCLUSIONS: Standard surgical treatment of varicose veins by saphenofemoral ligation, stripping and multiple phlebectomies is a clinically effective and cost-effective treatment for varicose veins, with an ICER well below the threshold normally considered appropriate for the funding of treatments within the NHS. Injection sclerotherapy also appears to be cost effective, but produces less overall benefit, with a higher ICER than surgery for patients with superficial venous reflux. In minor varicose veins without reflux, sclerotherapy is likely to provide a small average benefit with acceptable cost effectiveness. Research is needed into methods for accurate and acceptable utility evaluations for conditions with relatively minor effect on HRQoL and also for a validated and standardised method of classification for varicose veins. PMID- 16707071 TI - The cost-effectiveness of screening for oral cancer in primary care. AB - OBJECTIVES: To use a decision-analytic model to determine the incremental costs and outcomes of alternative oral cancer screening programmes conducted in a primary care environment. DESIGN: The cost-effectiveness of oral cancer screening programmes in a number of primary care environments was simulated using a decision analysis model. Primary data on actual resource use and costs were collected by case note review in two hospitals. Additional data needed to inform the model were obtained from published costs, from systematic reviews and by expert opinion using the Trial Roulette approach. The value of future research was determined using expected value of perfect information (EVPI) for the decision to screen and for each of the model inputs. SETTING: Hypothetical screening programmes conducted in a number of primary care settings. Eight strategies were compared: (A) no screen; (B) invitational screen--general medical practice; (C) invitational screen--general dental practice; (D) opportunistic screen--general medical practice; (E) opportunistic screen--general dental practice; (F) opportunistic high-risk screen--general medical practice; (G) opportunistic high-risk screen--general dental practice; and (H) invitational screen--specialist. PARTICIPANTS: A hypothetical population over the age of 40 years was studied. MAIN OUTCOME MEASURES: The main measures were mean lifetime costs and quality-adjusted life-years (QALYs) of each alternative screening scenario and incremental cost-effectiveness ratios (ICERs) to determine the additional costs and benefits of each strategy over another. RESULTS: No screening (strategy A) was always the cheapest option. Strategies B, C, E and H were never cost-effective and were ruled out by dominance or extended dominance. Of the remaining strategies, the ICER for the whole population (age 49-79 years) ranged from pound 15,790 to pound 25,961 per QALY. Modelling a 20% reduction in disease progression always gave the lowest ICERs. Cost-effectiveness acceptability curves showed that there is considerable uncertainty in the optimal decision identified by the ICER, depending on both the maximum amount that the NHS may be prepared to pay and the impact that treatment has on the annual malignancy transformation rate. Overall, however, high-risk opportunistic screening by a general dental or medical practitioner (strategies F and G) may be cost-effective. EVPIs were high for all parameters with population values ranging from pound 8 million to pound 462 million. However, the values were significantly higher in males than females but also varied depending on malignant transformation rate, effects of treatment and willingness to pay. Partial EVPIs showed the highest values for malignant transformation rate, disease progression, self-referral and costs of cancer treatment. CONCLUSIONS: Opportunistic high-risk screening, particularly in general dental practice, may be cost-effective. This screening may more effectively be targeted to younger age groups, particularly 40 60 year olds. However, there is considerable uncertainty in the parameters used in the model, particularly malignant transformation rate, disease progression, patterns of self-referral and costs. Further study is needed on malignant transformation rates of oral potentially malignant lesions and to determine the outcome of treatment of oral potentially malignant lesions. Evidence has been published to suggest that intervention has no greater benefit than 'watch and wait'. Hence a properly planned randomised controlled trial may be justified. Research is also needed into the rates of progression of oral cancer and on referral pathways from primary to secondary care and their effects on delay and stage of presentation. PMID- 16707072 TI - Measurement of the clinical and cost-effectiveness of non-invasive diagnostic testing strategies for deep vein thrombosis. AB - OBJECTIVES: To estimate the diagnostic accuracy of non-invasive tests for proximal deep vein thrombosis (DVT) and isolated calf DVT, in patients with clinically suspected DVT or high-risk asymptomatic patients, and identify factors associated with variation in diagnostic performance. Also to identify practical diagnostic algorithms for DVT, and estimate the diagnostic accuracy, clinical effectiveness and cost-effectiveness of each. DATA SOURCES: Electronic databases (to April 2004). A postal survey of hospitals in the UK. REVIEW METHODS: Selected studies were assessed against validated criteria. A postal survey of hospitals in the UK was undertaken to describe current practice and availability of tests, and identify additional diagnostic algorithms. Pooled estimates of sensitivity, specificity and likelihood ratios were obtained for each test using random effects meta-analysis. The effect of study-level covariates was explored using random effects metaregression. A decision-analytic model was used to combine estimates from the meta-analysis and estimate the diagnostic performance of each algorithm in a theoretical population of outpatients with suspected DVT. The net benefit of using each algorithm was estimated from a health service perspective, using cost--utility analysis, assuming thresholds of willingness to pay of pound 20,000 and pound 30,000 per quality-adjusted life-year (QALY). The model was analysed probabilistically and cost-effectiveness acceptability curves were generated to reflect uncertainty in estimated cost-effectiveness. RESULTS: Individual clinical features are of limited diagnostic value, with most likelihood ratios being close to 1. Wells clinical probability score stratifies proximal, but not distal, DVT into high-, intermediate- and low-risk categories. Unstructured clinical assessment by experienced clinicians may have similar performance to Wells score. In patients with clinically suspected DVT, D-dimer has 91% sensitivity and 55% specificity for DVT, although performance varies substantially between assays and populations. D-dimer specificity is dependent on pretest clinical probability, being higher in patients with a low clinical probability of DVT. Plethysmography and rheography techniques have modest sensitivity for proximal DVT, poor sensitivity for distal DVT, and modest specificity. Ultrasound has 94% sensitivity for proximal DVT, 64% sensitivity for distal DVT and 94% specificity. Computed tomography scanning has 95% sensitivity for all DVT (proximal and distal combined) and 97% specificity. Magnetic resonance imaging has 92% sensitivity for all DVT and 95% specificity. The diagnostic performance of all tests is worse in asymptomatic patients. The most cost-effective algorithm discharged patients with a low Wells score and negative D-dimer without further testing, and then used plethysmography alongside ultrasound, with venography in selected cases, to diagnose the remaining patients. However, the cost-effectiveness of this algorithm was dependent on assumptions of test independence being met and the ability to provide plethysmography at relatively low cost. Availability of plethysmography and venography is currently limited at most UK hospitals, so implementation would involve considerable reorganisation of services. Two algorithms were identified that offered high net benefit and would be feasible in most hospitals without substantial reorganisation of services. Both involved using a combination of Wells score, D-dimer and above-knee ultrasound. For thresholds of willingness to pay of pound 10,000 or pound 20,000 per QALY the optimal strategy involved discharging patients with a low or intermediate Wells score and negative D-dimer, ultrasound for those with a high score or positive D-dimer, and repeat scanning for those with positive D-dimer and a high Wells score, but negative initial scan. For thresholds of pound 30,000 or more a similar strategy, but involving repeat ultrasound for all those with a negative initial scan, was optimal. CONCLUSIONS: Diagnostic algorithms based on a combination of Wells score, D-dimer and ultrasound (with repeat if negative) are feasible at most UK hospitals and are among the most cost-effective. Use of repeat scanning depends on the threshold for willingness to pay for health gain. Further diagnostic testing for patients with a low Wells score and negative D-dimer is unlikely to represent a cost-effective use of resources. Recommendations for research include the evaluation of the costs and outcomes of using the optimal diagnostic algorithms in routine practice, the development and evaluation of algorithms appropriate for specific groups of patients with suspected DVT, such as intravenous drug abusers, pregnant patients and those with previous DVT, the evaluation of the role of plethysmography: interaction with other diagnostic tests, outcome of low-risk patients with negative plethysmography and measurement of the costs of providing plethysmography, and methodological research into the incorporation of meta analytic data into decision-analytic modelling. PMID- 16707073 TI - Systematic review of the effectiveness and cost-effectiveness of HealOzone for the treatment of occlusal pit/fissure caries and root caries. AB - OBJECTIVES: To assess the effectiveness and cost-effectiveness of HealOzone (CurOzone USA Inc., Ontario, Canada) for the management of pit and fissure caries, and root caries. The complete HealOzone procedure involves the direct application of ozone gas to the caries lesion on the tooth surface, the use of a remineralising solution immediately after application of ozone and the supply of a 'patient kit', which consists of toothpaste, oral rinse and oral spray all containing fluoride. DATA SOURCES: Electronic databases up to May 2004 (except Conference Papers Index, which were searched up to May 2002). REVIEW METHODS: A systematic review of the effectiveness of HealOzone for the management of tooth decay was carried out. A systematic review of existing economic evaluations of ozone for dental caries was also planned but no suitable studies were identified. The economic evaluation included in the industry submission was critically appraised and summarised. A Markov model was constructed to explore possible cost effectiveness aspects of HealOzone in addition to current management of dental caries. RESULTS: Five full-text reports and five studies published as abstracts met the inclusion criteria. The five full-text reports consisted of two randomised controlled trials (RCTs) assessing the use of HealOzone for the management of primary root caries and two doctoral theses of three unpublished randomised trials assessing the use of HealOzone for the management of occlusal caries. Of the abstracts, four assessed the effects of HealOzone for the management of occlusal caries and one the effects of HealOzone for the management of root caries. Overall, the quality of the studies was modest, with many important methodological aspects not reported (e.g. concealment of allocation, blinding procedures, compliance of patients with home treatment). In particular, there were some concerns about the choice of statistical analyses. In most of the full-text studies analyses were undertaken at lesion level, ignoring the clustering of lesions within patients. The nature of the methodological concerns was sufficient to raise doubts about the validity of the included studies' findings. A quantitative synthesis of results was deemed inappropriate. On the whole, there is not enough evidence from published RCTs on which to judge the effectiveness of ozone for the management of both occlusal and root caries. The perspective adopted for the study was that of the NHS and Personal Social Services. The analysis, carried out over a 5-year period, indicated that treatment using current management plus HealOzone cost more than current management alone for non-cavitated pit and fissure caries (40.49 pounds versus 24.78 pounds), but cost less for non-cavitated root caries ( 14.63 pounds versus 21.45 pounds). Given the limitations of the calculations these figures should be regarded as illustrative, not definitive. It was not possible to measure health benefits in terms of quality-adjusted life-years, due to uncertainties around the evidence of clinical effectiveness, and to the fact that the adverse events avoided are transient (e.g. pain from injection of local anaesthetic, fear of the drill). One-way sensitivity analysis was applied to the model. However, owing to the limitations of the economic analysis, this should be regarded as merely speculative. For non-cavitated pit and fissure caries, the HealOzone option was always more expensive than current management when the probability of cure using the HealOzone option was 70% or lower. For non-cavitated root caries the costs of the HealOzone comparator were lower than those of current management only when cure rates from HealOzone were at least 80%. The costs of current management were higher than those of the HealOzone option when the cure rate for current management was 40% or lower. One-way sensitivity analysis was also performed using similar NHS Statement of Dental Remuneration codes to those that are used in the industry submission. This did not alter the results for non-cavitated pit fissure caries as the discounted net present value of current management remained lower than that of the HealOzone comparator ( 22.65 pounds versus 33.39 pounds). CONCLUSIONS: Any treatment that preserves teeth and avoids fillings is welcome. However, the current evidence base for HealOzone is insufficient to conclude that it is a cost-effective addition to the management and treatment of occlusal and root caries. To make a decision on whether HealOzone is a cost-effective alternative to current preventive methods for the management of dental caries, further research into its clinical effectiveness is required. Independent RCTs of the effectiveness and cost-effectiveness of HealOzone for the management of occlusal caries and root caries need to be properly conducted with adequate design, outcome measures and methods for statistical analyses. PMID- 16707074 TI - Randomised controlled trials of conventional antipsychotic versus new atypical drugs, and new atypical drugs versus clozapine, in people with schizophrenia responding poorly to, or intolerant of, current drug treatment. AB - OBJECTIVES: To determine the clinical and cost-effectiveness of different classes of antipsychotic drug treatment in people with schizophrenia responding inadequately to, or having unacceptable side-effects from, their current medication. DESIGN: Two pragmatic, randomised controlled trials (RCTs) were undertaken. The first RCT (band 1) compared the class of older, inexpensive conventional drugs with the class of new atypical drugs in people with schizophrenic disorders, whose current antipsychotic drug treatment was being changed either because of inadequate clinical response or owing to side-effects. The second RCT (band 2) compared the new (non-clozapine) atypical drugs with clozapine in people whose medication was being changed because of poor clinical response to two or more antipsychotic drugs. Both RCTs were four-centre trials with concealed randomisation and three follow-up assessments over 1 year, blind to treatment. SETTING: Adult mental health settings in England. PARTICIPANTS: In total, 227 participants aged 18-65 years (40% of the planned sample) were randomised to band 1 and 136 (98% of the planned sample) to band 2. INTERVENTIONS: Participants were randomised to a class of drug. The managing clinician selected the individual drug within that class, except for the clozapine arm in band 2. The new atypical drugs included risperidone, olanzapine, quetiapine and amisulpride. The conventional drugs included older drugs, including depot preparations. As in routine practice, clinicians and participants were aware of the identity of the prescribed drug, but clinicians were asked to keep their participating patient on the randomised medication for at least the first 12 weeks. If the medication needed to be changed, the clinician was asked to prescribe another drug within the same class, if possible. MAIN OUTCOME MEASURES: The primary outcome was the Quality of Life Scale (QLS). Secondary clinical outcomes included symptoms [Positive and Negative Syndrome Scale (PANSS)], side-effects and participant satisfaction. Economic outcomes were costs of health and social care and a utility measure. RESULTS: Recruitment to band 1 was less than anticipated (40%) and diminished over the trial. This appeared largely due to loss of perceived clinical equipoise (clinicians progressively becoming more convinced of the superiority of new atypicals). Good follow-up rates and a higher than expected correlation between QLS score at baseline and at follow-up meant that the sample as recruited had 75% power to detect a difference in QLS score of 5 points between the two treatment arms at 52 weeks. The recruitment to band 2 was approximately as planned. Follow-up assessments were completed at week 52 in 81% of band 1 and 87% of band 2 participants. Band 1 data showed that, on the QLS and symptom measures, those participants in the conventional arm tended towards greater improvements. This suggests that the failure to find the predicted advantage for new atypicals was not due to inadequate recruitment and statistical power in this sample. Participants reported no clear preference for either class of drug. There were no statistically significant differential outcomes for participants entering band 1 for reasons of treatment intolerance to those entering because of broadly defined treatment resistance. Net costs over the year varied widely, with a mean of 18,850 pounds sterling in the conventional drug group and 20,123 pounds sterling in the new atypical group, not a statistically significant difference. Of these costs, 2.1% and 3.8% were due to antipsychotic drug costs in the conventional and atypical group, respectively. There was a trend towards participants in the conventional drug group scoring more highly on the utility measure at 1 year. The results for band 2 showed an advantage for commencing clozapine in quality of life (QLS) at trend level (p = 0.08) and in symptoms (PANSS), which was statistically significant (p = 0.01), at 1 year. Clozapine showed approximately a 5-point advantage on PANSS total score and a trend towards having fewer total extrapyramidal side-effects. Participants reported at 12 weeks that their mental health was significantly better with clozapine than with new atypicals (p < 0.05). Net costs of care varied widely, but were higher than in band 1, with a mean of 33,800 pounds sterling in the clozapine group and 28,400 pounds sterling in the new atypical group. Of these costs, 4.0% and 3.3%, respectively, were due to antipsychotic drug costs. The increased costs in the clozapine group appeared to reflect the licensing requirement for inpatient admission for commencing the drug. There was a trend towards higher mean participant utility scores in the clozapine group. CONCLUSIONS: For band 1, there is no disadvantage in terms of quality of life and symptoms, or associated costs of care, over 1 year in commencing conventional antipsychotic drugs rather than new atypical drugs. Conventional drugs were associated with non-significantly better outcomes and lower costs. Drug costs represented a small proportion of the overall costs of care (<5%). For band 2, there is a statistically significant advantage in terms of symptoms but not quality of life over 1 year in commencing clozapine rather than new atypical drugs, but with increased associated costs of care. The results suggest that conventional antipsychotic drugs, which are substantially cheaper, still have a place in the treatment of patients unresponsive to, or intolerant of, current medication. Further analyses of this data set are planned and further research is recommended into areas such as current antipsychotic treatment guidance, valid measures of utility in serious mental illness, low-dose 'conventional' treatment in first episode schizophrenia, QLS validity and determinants of QLS score in schizophrenia, and into the possible financial and other mechanisms of rewarding clinician participation in trials. PMID- 16707075 TI - Reproductive performance in infertile women with rectovaginal endometriosis: is surgery worthwhile? AB - OBJECTIVE: This study was undertaken to ascertain whether the incidence of pregnancy is increased and time-to-conception is reduced in infertile women with rectovaginal endometriosis undergoing conservative surgery compared with those on expectant management. STUDY DESIGN: A total of 105 infertile women under the age of 40 years with rectovaginal endometriosis and no other associated major infertility factor underwent first-line conservative surgery at laparotomy or expectant management according to a shared decision-making approach. RESULTS: Among the 44 women who had resection of rectovaginal endometriosis, 15 became pregnant, compared with 22 of the 61 women who choose expectant management (24 month cumulative probabilities, 44.9% and 46.8%, respectively; log-rank test, chi2(1) = 0.75; P = .38). One major and 9 minor postoperative complications occurred. Significant differences in pain-free survival time in favor of the surgery group were observed for dysmenorrhea, dyspareunia, and dyschezia. CONCLUSION: Conservative surgery for rectovaginal endometriosis in infertile women does not modify the reproductive prognosis although it does increase pain free survival time. PMID- 16707076 TI - Placental transfer of rosiglitazone in the ex vivo human perfusion model. AB - OBJECTIVE: The objective of the study was to determine transplacental passage of rosiglitazone (Avandia) using the ex vivo human placental model. STUDY DESIGN: Perfusion studies were performed on 10 placentas from term, uncomplicated deliveries. Concentrations typical for an 8-mg oral dose (216 to 692 ng/mL) as well as 2- to 3-fold increased concentrations were tested (734 to 1261 ng/mL). Transfer of rosiglitazone was assessed and accumulation was determined using the 14C-antipyrine reference method. RESULTS: The clearance index for low and high concentrations were 0.14 +/- 0.04 and 0.20 +/- 0.08, suggesting that the drug passes through the placenta at a relatively low rate. Fetal accumulation occurred in only 1 of 5 placentas at 16.4 ng/mL (5%) for the 8-mg dose and in 2 of 5 placentas ranging from 0 to 74 ng/mL (5% to 8%) at higher concentrations. CONCLUSION: There is minimal transfer and fetal accumulation of rosiglitazone according to the ex vivo human perfusion model. PMID- 16707077 TI - Neonatal neurodevelopmental outcomes following tocolysis with glycerol trinitrate patches. AB - OBJECTIVE: The object of this study was to determine the effects of maternal tocolysis with glycerol trinitrate (GTN) patches on the neurodevelopment of infants. STUDY DESIGN: This was a randomized, multicenter, controlled trial comparing the efficacy of GTN patches with standard beta2 agonist as tocolytic therapy. The previously reported outcomes of this study indicated no difference in neonatal mortality or morbidity to hospital discharge. One hundred fifty-six surviving infants from 2 Australian centers were psychometrically assessed using the Griffiths Mental development Scales (revised) at 18 months of age. RESULTS: There was no difference in psychometric performance between those infants enrolled in either the GTN (81 infants) or beta2 agonist (75 infants) arm of the study. CONCLUSION: This randomized trial supports no significant difference between GTN patches in comparison with standard beta2 agonist for tocolytic therapy. The results underscore the association between premature labor and adverse infant outcomes. PMID- 16707078 TI - Fetal transcerebellar diameter measurement for prediction of gestational age in twins. AB - OBJECTIVE: This study was undertaken to determine the accuracy of our previously published and prospectively validated institution-specific singleton transcerebellar diameter (TCD) nomogram in the prediction of gestational age (GA) in twin pregnancies. We further evaluated whether the prediction of GA in twin gestations using the singleton TCD nomogram differs between monochorionic and dichorionic twins. STUDY DESIGN: In our previously published studies, we retrospectively constructed a cross-sectional nomogram using TCD measurements in 24,026 well-dated, singleton fetuses, and prospectively validated the nomogram using 2,597 singleton fetuses. The current study comprised of 1,278 well-dated twins (19.6% monochorionic) seen in our ultrasound unit between August 1994 and May 2003, and the singleton TCD nomogram was validated in these twin gestations. The actual GA was subtracted from the GA predicted by the TCD nomogram and the concordance between actual and predicted GAs was assessed on the basis of the Pearson's correlation coefficient (r). This was performed separately for monochorionic and dichorionic twins. RESULTS: Concordance between the actual and predicted twin TCD measurements based on our previously published singleton TCD nomogram was high (Pearson's correlation, r = 0.95, P < .0001). Between 16 and 23 weeks' gestation, the predicted mean GA was within 6 days of actual GA. Between 24 and 30 weeks, the predicted mean GA was within 3 days, and at 32 weeks or more, the predicted mean GA was within 5 days of the actual GA. Prediction of GA based on the singleton TCD nomogram was equally accurate in both monochorionic and dichorionic twin gestations (P = .686). CONCLUSION: This study demonstrates that our previously validated singleton TCD nomogram is reliable and accurate in twins irrespective of placental chorionicity. PMID- 16707079 TI - Extremity amniotic band syndrome in fetal lamb. I: An experimental model of limb amputation. AB - OBJECTIVE: The objective of the study was to reproduce severe forms of extremity amniotic bands, which result in amputation or severe lesions. STUDY DESIGN: Right limbs of 5 60-day gestational age pregnant sheep were ligated with silk suture at the infracondylar level. Left limbs were used for paired comparison. The limbs obtained from term fetuses were analyzed morphologically, radiologically, and histologically. RESULTS: The ligated limbs showed an amputation or quasiamputation. Four extremities showed a necrotic pattern and 4 an edema pattern. CONCLUSION: A model of severe extremity amniotic bands that reproduces the effect of the extremity amniotic bands in the human fetus was developed. Future applications of this new model could be an experimental study of in utero salvage of limbs affected with severe extremity amniotic bands in the ovine fetus. PMID- 16707081 TI - Characteristics of women giving birth at home in Sweden: a national register study. AB - OBJECTIVE: The objective of the study was to estimate the proportion of planned home births in Sweden and to identify maternal characteristics of women giving birth at home. STUDY DESIGN: This case-control study included register data of births from 1992 to 2001 in 352 women giving birth at home and 1760 women giving birth in a hospital. RESULTS: Four hundred thirty-nine out-of-hospital births were found during the study period, and the proportion of planned home births was less than 0.5/1000. Women with home birth were more likely to have 4 children or more (odds ratio 3.7 [1.4 to 9.9]), be born in a European country outside Sweden (odds ratio 3.5 [1.8 to 6.8]), have a family income below the median (odds ratio 2.9 [2.0 to 4.1]), not work outside the home (odds ratio 2.4 [1.7 to 3.5]), have a high level of education (odds ratio 2.1 [1.5 to 3.0]), and be older than 35 years (odds ratio 1.7 [1.1 to 2.5]). CONCLUSION: Women with planned home births appear to be a group having a different lifestyle, compared with Swedish women in general. PMID- 16707082 TI - The role of laser surgery in dissecting the etiology of absent or reverse end diastolic velocity in the umbilical artery of the donor twin in twin-twin transfusion syndrome. AB - OBJECTIVE: This study was undertaken to gain insight on the cause of absent or reverse end-diastolic velocity (AREDV) in the umbilical artery (UA) of the donor twin by analysis of individual placental mass and vascular anastomoses in patients with twin-twin transfusion syndrome (TTTS) treated with laser. STUDY DESIGN: TTTS patients who successfully underwent selective laser photocoagulation of communicating vessels (SLPCV), 16 and 26 weeks' gestation, with both twins born alive and complete Doppler and placental data were considered eligible for the study. Doppler examination of the UA was performed before and 24 hours after SLPCV. Abnormal UA Doppler findings were defined as persistent AREDV. Pre- and post-SLPCV UA Doppler results yielded the following 4 groups: (1) normal-normal; (2) normal-abnormal; (3) abnormal-normal; and (4) abnormal-abnormal. The types of vascular anastomoses were categorized during surgery. Individual placental territory (IPT) was defined as individual placental weight divided by total placental weight x 100. RESULTS: There were 132 cases in group 1 and no patients in group 2. AREDV resolved in 78% (28/36) of patients (group 3) and remained unchanged in 22% (8/36) (group 4). The mean IPT-donor in group 4 was significantly smaller than in group 1 (P = .015). Patients with preoperative AREDV (groups 3 and 4) were more likely to have artery-to-artery anastomoses (P = .002). However, AREDV resolved in 57% (16/28) patients without artery-to-artery anastomoses. CONCLUSION: Preoperative AREDV may result from a small IPT, placental vascular anastomoses, or both. AREDV resulting from a small IPT may have a similar cause to that of singletons, and may be inferred by lack of postoperative resolution. Resolution of AREDV after SLPCV implies the presence of an adequate IPT and removal of donor hypotension. PMID- 16707085 TI - Voltammetric determination of mefenamic acid at lanthanum hydroxide nanowires modified carbon paste electrodes. AB - Lanthanum hydroxide nanowires modified carbon paste electrode (LNW/CPE) exhibiting an electrocatalytic response toward the oxidation of mefenamic acid (MFA) is described. The catalytic action of the LNW/CPE on the oxidation of MFA via one-electron and one-proton transfer is attributed to the formation of the porous construction and the increase of efficient surface of the electrode due to the adulteration of LNW with carbon powders. Using the LNW/CPE, a linear sweep voltammetric method for the determination of MFA and other drugs with diphenylamine parent is proposed. A linear range of 2.0 x 10(-11) to 4.0 x 10( 9)mol L(-1) is obtained along with a detection limit of 6.0 x 10(-12)mol L(-1). PMID- 16707084 TI - A rapid and efficient method for purification of recombinant adenovirus with arginine-glycine-aspartic acid-modified fibers. AB - Recombinant adenoviral vectors (adenovectors) have been subject to various genetic modifications to improve their transduction efficiency and targeting capacity. Production and purification of adenovectors with modified capsid proteins can be problematic using conventional two-cycle CsCl gradient ultracentrifugation. We have developed a new method for purifying recombinant adenovectors in two steps: iodixanol discontinuous density gradient ultracentrifugation and size exclusion column chromatography. The purity and infectious activity of adenovectors isolated by the two methods were comparable. The new method yielded three to four times more adenovectors with arginine glycine-aspartic acid (RGD)-modified fiber proteins than did the conventional CsCl method. For other fiber-modified and wild-type adenovectors, the yields of the two methods were comparable. Thus, the iodixanol-based method can be used not only to improve the production of RGD-modified adenovectors but also to purify adenovectors with or without fiber modifications. Moreover, the whole procedure can be completed in 3h. Therefore, this method is rapid and efficient for production of recombination adenovectors, especially those with RGD-modified fibers. PMID- 16707086 TI - A novel scintillation proximity assay for fatty acid amide hydrolase compatible with inhibitor screening. AB - A binding assay for human fatty acid amide hydrolase (FAAH) using the scintillation proximity assay (SPA) technology is described. This SPA uses the specific interactions of [3H]R(+)-methanandamide (MAEA) and FAAH expressing microsomes to evaluate the displacement activity of FAAH inhibitors. We observed that a competitive nonhydrolyzed FAAH inhibitor, [3H]MAEA, bound specifically to the FAAH microsomes. Coincubation with an FAAH inhibitor, URB-597, competitively displaced the [3H]MAEA on the FAAH microsomes. The released radiolabel was then detected through an interaction with the SPA beads. The assay is specific for FAAH given that microsomes prepared from cells expressing the inactive FAAH-S241A mutant or vector alone had no significant ability to bind [3H]MAEA. Furthermore, the binding of [3H]MAEA to FAAH microsomes was abolished by selective FAAH inhibitors in a dose-dependent manner, with IC50 values comparable to those seen in a functional assay. This novel SPA has been validated and demonstrated to be simple, sensitive, and amenable to high-throughput screening. PMID- 16707087 TI - Electrochemical DNA biosensor for the study of ciprofloxacin-DNA interaction. AB - The interaction of ciprofloxacin with DNA was studied by using an electrochemical DNA biosensor. The binding mechanism of ciprofloxacin was elucidated by using constant current potentiometry and differential pulse voltammetry at DNA-modified glassy carbon electrode. The decrease in the guanine oxidation peak area or peak current at +0.9 V was used as an indicator for the interaction mechanism in 0.2M acetate buffer (pH 5). The binding constant (K) values obtained were 1.33+/-0.02 x 10(4) and 1.32+/-0.08 x 10(4) M(-1) with constant current potentiometry and differential pulse voltammetry, respectively. A linear dependence of the guanine peak area or peak currents was observed in the range of 40-80 microM ciprofloxacin, with a detection limit of 24 microM with r=0.995 and 9 microM with r=0.999 by using constant current potentiometry and differential pulse voltammetry, respectively. Moreover, the influence of sodium and calcium ions was also studied to elucidate the mechanism of ciprofloxacin-DNA interaction at different solution conditions, and this proved to be helpful in understanding the ciprofloxacin-DNA interaction. PMID- 16707088 TI - Simultaneous liquid chromatography determination of polyamines and arylalkyl monoamines. AB - The diamines putrescine (PUT) and diaminopropane (DAP), the polyamines spermidine (SPD) and spermine (SPM), and the arylalkyl amines phenethylamine (PEA), tyramine (TYR), dopamine (DA), and salsolinol (SAL) were dansylated and baseline separated by LC using a Waters ODS-2 column. The dansyl derivatives were detected by fluorescence (lambda(ex): 337 nm; lambda(em): 520 nm). Besides the amine function, the phenolic OH groups of TYR, DA, and SAL were also dansylated (LC-MS, formation of N,O-didansyl [TYR] and N,O,O'-tridansyl derivatives [DA and SAL]). Calibration curves revealed response factors being appreciably lower for (N,O didansyl) aminophenol TYR and (N,O,O'-tridansyl) DA and SAL than for N dansylamines. However, the method is suitable as a cheap alternative to LC-MS for the simultaneous determination of polyamines and arylalkyl amines of large quantities of samples. PMID- 16707089 TI - A high-throughput screening assay for the carboxyltransferase subunit of acetyl CoA carboxylase. AB - One consequence of the dramatic rise of antibiotic-resistant pathogenic bacteria is the need for new targets for antibiotics. Because membrane lipid biogenesis is essential for bacterial growth, enzymes of the fatty acid biosynthetic pathway offer attractive possibilities for the development of new antibiotics. Acetyl coenzyme A carboxylase (ACC) catalyzes the first committed and regulated step in fatty acid biosynthesis in bacteria and thus is a prime target for development of antibiotics. ACC is a multifunctional enzyme composed of three separate proteins. The biotin carboxylase component catalyzes the ATP-dependent carboxylation of biotin. The biotin carboxyl carrier protein features a biotin molecule covalently attached at Lys122 of the Escherichia coli enzyme. The carboxyltransferase subunit catalyzes the transfer of a carboxyl group from biotin to acetyl-coenzyme A (acetyl-CoA) to form malonyl-CoA. The objective of this study was to develop an assay for high-throughput screening for inhibitors of the carboxyltransferase subunit. The carboxyltransferase reaction was assayed in the reverse direction in which malonyl-CoA reacts with biocytin (an analog of the biotin carboxyl carrier protein) to form acetyl-CoA and carboxybiotin. The production of acetyl-CoA was coupled to citrate synthase, which produced citrate and coenzyme A. The amount of coenzyme A formed was detected using 5,5'-dithiobis(2-nitrobenzoic acid) (Ellman's reagent). The assay has been developed for use in both 96- and 384-well microplate formats and was validated using a known bisubstrate analog inhibitor of carboxyltransferase. The spectrophotometric readout in the visible absorbance range used in this assay does not generate the number of false negatives associated with frequently used NAD/NADH assay systems that rely on detection of NADH using UV absorbance. PMID- 16707090 TI - Microwave-assisted spectrophotometric estimation of functional groups on biochips using a universal reagent. PMID- 16707091 TI - In vitro selection of translational regulatory elements. AB - Untranslated regions (UTRs) of mRNAs carry various kinds of translational regulatory elements; however, our knowledge of them is still limited. We created an in vitro selection system that allows us to make a systematic enrichment of the sequences that alter translation efficiency (SESTRE) in any given mRNA and translation system. This method consists of the introduction of random nucleotide sequences into the UTRs of given mRNAs, followed by translation, size fractionation of the polyribosomes, and reverse transcription and PCR amplification (RT-PCR), with repeated cycles of these steps to enrich highly or poorly translatable mRNAs. With this experimental method, we examined how and where translational enhancer motifs emerge on mRNAs using the in vitro translation systems of wheat germ extract. The results indicate that the translational enhancers differentially emerge in response to the presence or absence of the 5' cap. Interestingly, the translational enhancers that activate cap-independent translation evolved more readily in the 3' UTR than in the 5' UTR in wheat germ extract. This SESTRE method should be a powerful tool with which to improve the translational efficiency of given mRNAs in given translation systems and to investigate the structure-function relationship of eukaryotic mRNAs underlying translational control. PMID- 16707092 TI - Measurement of carnitine biosynthesis enzyme activities by tandem mass spectrometry: differences between the mouse and the rat. AB - Although the mouse frequently is used to study metabolism and deficiencies therein, little is known about carnitine biosynthesis in this animal. To this point, only laborious procedures have been described to measure the activity of carnitine biosynthesis enzymes using subcellular fractions as the enzyme source. We developed two simple tandem mass spectrometry-based methods to determine the activity of three carnitine biosynthesis enzymes (6-N-trimethyllysine dioxygenase, 4-trimethylaminobutyraldehyde dehydrogenase, and 4 trimethylaminobutyric acid dioxygenase) in total homogenates that can be prepared from frozen tissue. The new assays were used to characterize these enzymes in mouse liver homogenate. Because carnitine biosynthesis has been studied extensively in the rat, we compared the mouse tissue distribution of carnitine biosynthesis enzyme activities and levels of the biosynthesis metabolites with those in the rat to determine which tissues contribute to carnitine biosynthesis in these species. Surprisingly, large differences in enzyme activities were found between the rat and the mouse, whereas carnitine biosynthesis metabolite levels were very similar in both species, possibly due to the different kinetic properties of the first enzyme of carnitine biosynthesis. Also, muscle carnitine levels were found to vary considerably between these two species, suggesting that there is a metabolic dissimilarity between the mouse and the rat. PMID- 16707093 TI - [Pelvic posterior exenteration with immediate colo-rectal anastomosis: is it justified and feasible in advanced stage ovarian carcinoma?]. AB - PURPOSE: The aim of this study is to show that the removal of the rectum is not an obstacle to implement an optimal surgery in advanced epithelial cancer of the ovary. MATERIAL AND METHODS: Retrospective study on a population of 44 women with advanced epithelial cancer of the ovary. The surgery was realized between January 95 and July 03, and all surgeries required a posterior exenteration. This treatment was completed by chemotherapy for 36 of them. RESULTS: The median survival of this population is 36.6 months. 6/44 patients (13.6%) had post operative complications. The completion of chemotherapy started after an average of 5.2 weeks after surgery. All the assessable patients (43/44) have an anal satisfactory continence. CONCLUSION: The posterior exenteration, when it's necessary, for advanced epithelial cancer of the ovary must not be an obstacle to obtain an optimal surgery. Anal continence is respected and there are no more complications. This surgical act is safe for the management of this pathology without delaying the others therapeutics and allowing a satisfactory quality of life. PMID- 16707094 TI - [Abbreviated laparotomy in severe abdominal trauma]. AB - On critically injured patient the decision to perform a damage control laparotomy is based on the volume of transfusion and shock. The aim of the surgery which is to obtain as fast as possible the best hemostasis to limit the peritoneal thermal loss and to perform as soon as possible physiologic restoration in the Intensive Care Unit. PMID- 16707095 TI - Increased level of DJ-1 in the cerebrospinal fluids of sporadic Parkinson's disease. AB - DJ-1 is an antioxidant protein whose loss of function by gene mutations has been linked to familial Parkinson's disease (PD). The main objective of the present study was to determine if this molecule was also involved in the pathogenesis of sporadic PD. For this purpose, quantitative immunoblot assays were performed to evaluate DJ-1 in cerebrospinal fluids (CSF) collected from sporadic PD patients (n=40) and non-PD controls (n=38). The results showed that the CSF DJ-1 levels in PD were significantly higher than those in non-PD controls. Especially, upregulation of CSF DJ-1 in the early stage of PD (Yahr I-II) were distinct compared to those in the advanced stage of PD (Yahr III-IV) and non-PD controls (p<0.001 by ANOVA with post hoc Bonferroni's test), suggesting a protective role of DJ-1 against oxidative stress during the early stage. Thus, we propose that CSF DJ-1 could be a possible biomarker for early sporadic PD. PMID- 16707096 TI - Vasohibin prevents arterial neointimal formation through angiogenesis inhibition. AB - Vasohibin is a VEGF-inducible angiogenesis inhibitor in vascular endothelium. Here we examined the presence of vasohibin in human arterial wall, and found it in endothelium of adventitial microvessels in atherosclerotic lesion. Adventitial angiogenesis is involved in the progression of neointimal formation. Even in the presence of endogenous angiogenesis inhibitors, pathological angiogenesis persists. However, the supplementation of exogenous angiogenesis inhibitors can prevent pathological angiogenesis. We evaluated the potential role of vasohibin in neointimal formation. Adenovirus-mediated human vasohibin gene transfer in mouse liver resulted in the release of vasohibin in plasma and exhibited anti angiogenic effects at remote sites. This gene transfer inhibited adventitial angiogenesis, macrophage infiltration, and neointimal formation after cuff placement on mouse femoral artery. Vasohibin exhibited no direct effect on migration and proliferation of smooth muscle cells. Thus, vasohibin has an activity to prevent neointimal formation by inhibiting adventitial angiogenesis. PMID- 16707097 TI - Hydrogen sulfide potentiates interleukin-1beta-induced nitric oxide production via enhancement of extracellular signal-regulated kinase activation in rat vascular smooth muscle cells. AB - Hydrogen sulfide (H(2)S) and nitric oxide (NO) are endogenously synthesized from l-cysteine and l-arginine, respectively. They might constitute a cooperative network to regulate their effects. In this study, we investigated whether H(2)S could affect NO production in rat vascular smooth muscle cells (VSMCs) stimulated with interleukin-1beta (IL-1beta). Although H(2)S by itself showed no effect on NO production, it augmented IL-beta-induced NO production and this effect was associated with increased expression of inducible NO synthase (iNOS) and activation of nuclear factor (NF)-kappaB. IL-1Beta activated the extracellular signal-regulated kinase 1/2 (ERK1/2), and this activation was also enhanced by H(2)S. Inhibition of ERK1/2 activation by the selective inhibitor U0126 inhibited IL-1beta-induced NF-kappaB activation, iNOS expression, and NO production either in the absence or presence of H(2)S. Our findings suggest that H(2)S enhances NO production and iNOS expression by potentiating IL-1beta-induced NF-kappaB activation through a mechanism involving ERK1/2 signaling cascade in rat VSMCs. PMID- 16707098 TI - Upregulation of aspartoacylase activity in the duodenum of obesity induced diabetes mouse: implications on diabetic neuropathy. AB - Aspartoacylase (ASPA) hydrolyzes N-acetylaspartic acid (NAA) into aspartate and acetate. Normal hydrolysis of NAA is important to maintain healthy neurons. Since enteric neuropathy is one of the events seen in diabetes, whether ASPA activity is affected in diabetic condition is not known. In order to investigate the possibility, ASPA activity was examined in the duodenum and brain of obesity induced diabetes model mouse. Aspartoacylase activity was high in the diabetic mouse duodenum compared to control duodenum. The same result was also observed by immunostaining of the mouse duodenum. The activity of ASPA was found to be elevated in the brain of diabetic mouse compared to the control brain. These data suggest that normal hydrolysis of NAA is affected by ASPA activity seen in the type 2 diabetes model mouse and this change is likely to contribute to neuropathy seen in diabetes, if documented also in patients with type 2 diabetes. PMID- 16707099 TI - Efficient establishment of human embryonic stem cell lines and long-term maintenance with stable karyotype by enzymatic bulk passage. AB - Human ES (hES) cell lines are considered to be a valuable resource for medical research and for applications in cell therapy and drug discovery. For such utilization of hES cells to be realized, however, protocols involved in the use of hES cells, such as those for establishment, propagation, and cryopreservation, have still to be improved. Here, we report on an efficient method for the establishment of hES cell lines and its detailed characterization. Additionally, we developed a new bulk-passaging technique that preserves the karyotypic integrity of hES cell lines when maintained in culture for up to 2 years. Finally, we show that a simplified vitrification cryopreservation technique is vastly superior to standard slow-cooling methods with respect to cell viability. These results provide valuable information that will assist in achieving the goal of the large-scale hES cell culture required for the application of hES cells to disease therapy. PMID- 16707100 TI - Characterization of calcineurin-dependent response element binding protein and its involvement in copper-metallothionein gene expression in Neurospora. AB - In continuation of our recent observations indicating the presence of a lone calcineurin-dependent response element (CDRE) in the -3730bp upstream region of copper-induced metallothionein (CuMT) gene of Neurospora [K.S. Kumar, S. Dayananda, C. Subramanyam, Copper alone, but not oxidative stress, induces copper metallothionein gene in Neurospora crassa, FEMS Microbiol. Lett. 242 (2005) 45 50], we isolated and characterized the CDRE-binding protein. The cloned upstream region of CuMT gene was used as the template to specifically amplify CDRE element, which was immobilized on CNBr-activated Sepharose 4B for use as the affinity matrix to purify the CDRE binding protein from nuclear extracts obtained from Neurospora cultures grown in presence of copper. Two-dimensional gel electrophoresis of the affinity purified protein revealed the presence of a single 17kDa protein, which was identified and characterized by MALDI-TOF. Peptide mass finger printing of tryptic digests and analysis of the 17kDa protein matched with the regulatory beta-subunit of calcineurin (Ca(2+)-calmodulin dependent protein phosphatase). Parallel identification of nuclear localization signals in this protein by in silico analysis suggests a putative role for calcineurin in the regulation of CuMT gene expression. PMID- 16707101 TI - Correspondence regarding Schwend and Gustafsson, "False positives in MALDI-TOF detection of ERbeta in mitochondria". AB - Recently, Schwend and Gustafsson tried to use the MALDI-TOF methods to confirm one of the results reported by Yang et al., which provided definitive evidences to demonstrate the localization of estrogen receptor beta (ERbeta) in the mitochondria of multiple cell types, using immunocytochemistry, immunoblot, and proteomic approaches. Analysis of the data with the MASCOT database algorithm provided no evidence for the presence of ERbeta in the mouse live mitochondria, in which very low ERbeta expression has been detected in their own report. On the other hand, our MALDI-TOF analysis using human heart mitochondrial protein has identified 7 and 8 sequences that could be potentially from ERbeta and ERbeta3, respectively, but not from ATP synthases. Further, none of the sequences identified by us as those of ERbeta and ERbeta3 shares m/z targeted by Schwend and Gustafsson in their measurements. Therefore, the claim by Gustafsson's laboratory about false positives in MALDI-TOF detection of ERbeta in mitochondria has no relevance to our report. PMID- 16707102 TI - Establishment of primary cultures for mouse ameloblasts as a model of their lifetime. AB - To understand how the properties of ameloblasts are spatiotemporally regulated during amelogenesis, two primary cultures of ameloblasts in different stages of differentiation were established from mouse enamel epithelium. Mouse primary ameloblasts (MPAs) prepared from immature enamel epithelium (MPA-I) could proliferate, whereas those from mature enamel epithelium (MPA-M) could not. MPA-M but not MPA-I caused apoptosis during culture. The mRNA expression of amelogenin, a marker of immature ameloblasts, was down-regulated, and that of enamel matrix serine proteiase-1, a marker of mature ameloblasts, was induced in MPA-I during culture. Using green fluorescence protein as a reporter, a visualized reporter system was established to analyze the promoter activity of the amelogenin gene. The region between -1102bp and -261bp was required for the reporter expression in MPA-I. These results suggest that MPAs are valuable in vitro models for investigation of ameloblast biology, and that the visualized system is useful for promoter analysis in MPAs. PMID- 16707103 TI - NMR structure of an intracellular loop peptide derived from prostaglandin EP3alpha receptor. AB - We found that a peptide (EP3a: TIKALVSRCRAKAAV) corresponding to the N-terminal site of the intracellular third loop of human prostaglandin EP3alpha receptor could activate G protein alpha-subunit directly. The activity was almost same as Mastoparan-X, a G protein activating peptide from wasp venom. The three dimensional molecular structure of the peptide in SDS-d(25) micelles was determined by 2D (1)H NMR spectroscopy. The structure of EP3a consists of a positive charge cluster on the C-terminal helical site. The cluster was also found in several corresponding receptor peptides. Therefore, the positive charge cluster on the helical structure might play a crucial role in activation of G protein. PMID- 16707104 TI - Fusarium graminearum on plant cell wall: no fewer than 30 xylanase genes transcribed. AB - The transcription of a set of 32 putative xylanase genes from Fusarium graminearum was examined by quantitative PCR after growth on different carbon sources (hop cell wall, xylan, xylose, or carboxymethylcellulose). Growing on plant cell wall medium, this fungus displays a great diversity of expression of xylan-related genes, with 30 being induced. A second level of diversity exists because expression patterns can be very different for loci encoding enzymes with the same activity (the same EC number). The wealth of xylan-degrading enzymes and the differential expression confer on the fungus a great flexibility of reaction to variation in its environment. PMID- 16707105 TI - Improvement of diabetes, obesity and hypertension in type 2 diabetic KKAy mice by bis(allixinato)oxovanadium(IV) complex. AB - Previously, we found that bis(allixinato)oxovanadium(IV) (VO(alx)(2)) exhibits a potent hypoglycemic activity in type 1-like diabetic mice. Since the enhancement of insulin sensitivity is involved in one of the mechanisms by which vanadium exerts its anti-diabetic effects, VO(alx)(2) was further tested in type 2 diabetes with low insulin sensitivity. The effect of oral administration of VO(alx)(2) was examined in obesity-linked type 2 diabetic KKA(y) mice. Treatment of VO(alx)(2) for 4 weeks normalized hyperglycemia, glucose intolerance, hyperinsulinemia, hypercholesterolemia and hypertension in KKA(y) mice; however, it had no effect on hypoadiponectinemia. VO(alx)(2) also improved hyperleptinemia, following attenuation of obesity in KKA(y) mice. This is the first example in which a vanadium compound improved leptin resistance in type 2 diabetes by oral administration. On the basis of these results, VO(alx)(2) is proposed to enhance not only insulin sensitivity but also leptin sensitivity, which in turn improves diabetes, obesity and hypertension in an obesity-linked type 2 diabetic animal. PMID- 16707106 TI - Activity-dependent regulation of beta-catenin via epsilon-cleavage of N-cadherin. AB - N-cadherin is essential for excitatory synaptic contact in the hippocampus. Presenilin 1 (PS1) is located at sites of synaptic contact, forming a complex with N-cadherin and beta-catenin. Here, we report that human N-cadherin is cleaved by PS1/gamma-secretase in response to physiological concentration of glutamate (Glu) stimulation, yielding a fragment Ncad/CTF2. The expression of Ncad/CTF2 in neuronal cells led to its translocation to the nucleus, and caused a prominent enhancement of cytoplasmic and nuclear beta-catenin levels in a cell cell contact dependent manner, via following mechanisms: 1, inhibition of beta catenin phosphorylation; 2, transactivation of beta-catenin; and 3, inhibition of N-cadherin transcription, and finally enhanced beta-catenin nuclear signaling. Since the regulation of cellular beta-catenin level is essential for synaptic function, disruption in the cleavage of N-cadherin may be causally linked to the synaptic dysfunction associated with Alzheimer's disease (AD). PMID- 16707107 TI - Endocrine disrupting chemicals (bisphenol A, 4-nonylphenol, 4-tert-octylphenol) modulate expression of two distinct cytochrome P450 aromatase genes differently in gender types of the hermaphroditic fish Rivulus marmoratus. AB - To understand the effect of endocrine-disrupting chemicals (EDCs) on cytochrome P450 aromatase (rm-cyp19) gene expression between gender types in the hermaphroditic fish Rivulus marmoratus, we cloned two distinct rm-cyp19 genes using RT-PCR with degenerative primers, obtained full-length cDNAs using 5'- and 3'-RACE-PCR methods, and completely sequenced them. The brain aromatase (rm cyp19b) cDNA consisted of 2,124 bp including the open reading frame (ORF), which encoded a putative protein of 505 amino acids. The ovarian aromatase (rm-cyp19a) cDNA consisted of 2,075 bp, including the ORF encoding a putative protein of 516 amino acids. Expression patterns of rm-cyp19b and rm-cyp19a mRNAs were investigated in embryos of different developmental stages and in seven different tissues of adult fish. The rm-cyp19b gene in hermaphrodite and secondary male R. marmoratus was predominantly expressed in the brain, while the rm-cyp19a gene was expressed gender-specifically in the gonad. The expression of rm-cyp19b mRNA increased from stage 1 (2 d post fertilization) to stage 4 (12 d post fertilization) in a developmental stage-dependent manner but steeply decreased in the hatching stage. Compared to the rm-cyp19b gene, the abundance of ovarian aromatase rm-cyp19a transcripts was very low, and its expression was first detected at stage 3 and then decreased gradually to the hatching stage. Alteration of rm-cyp19b and rm-cyp19a gene expression was further analyzed in the brain and gonad by real-time RT-PCR 96 h after EDC exposure in hermaphrodites and secondary males. The brain aromatase rm-cyp19b gene was up-regulated in the brain after 4-nonylphenol (4-NP)-exposure, while the ovarian aromatase rm-cyp19a gene was significantly down-regulated in the gonad. In 300 microg/L 4-tert octylphenol (4-tert-OP), or 600 microg/L bisphenol A-exposed brain and gonad, both rm-cyp19b and rm-cyp19a genes were up-regulated. In the case of secondary males, the rm cyp19b gene was highly expressed in the 4-NP-exposed brain, while expression of the rm-cyp19a gene was not detected in the gonad. These results indicate that the expression of rm-cyp19a and rm-cyp19b genes is differently modulated according to estrogenic compounds and gender type of R. marmoratus. PMID- 16707108 TI - JSAP1 is required for the cell adhesion and spreading of mouse embryonic fibroblasts. AB - The roles of JSAP1 and JIP1 in cell adhesion and spreading were examined using mouse embryonic fibroblasts (MEFs) deficient in JIP1 (JIP1-KO), JSAP1 (JSAP1-KO), and in both JIP1 and JSAP1 (double-KO), and by using their wild type. After being plated on fibronectin-coated culture plates, wild type MEFs rapidly adhered and differentiated to typical longitudinal fibroblasts in 4 h. JSAP1-KO MEFs showed a similar sequence of adhesion and cell spreading, but their adhesion was weak, and cell spreading sequence proceeded in a delayed manner compared with the wild type. In spreading JSAP1-KO MEFs, adhesion-triggered actin cytoskeleton reorganization and FAK activation proceeded at a slower pace than in wild type MEFs. The cellular properties of double-KO MEFs and JIP1-KO MEFs were similar to those of JSAP1-KO MEFs and wild type MEFs, respectively. These results suggest that JSAP1 plays a role in adhesion and cell spreading by regulating the rapid reorganization of the actin cytoskeleton. PMID- 16707109 TI - wnt3a but not wnt11 supports self-renewal of embryonic stem cells. AB - wnt proteins (wnts) promote both differentiation of midbrain dopaminergic cells and self-renewal of haematopoietic stem cells. Mouse embryonic stem (ES) cells can be maintained and self-renew on mouse feeder cell layers or in media containing leukemia inhibitory factor (LIF). However, the effects of wnts on ES cells self-renewal and differentiation are not clearly understood. In the present study, we found that conditioned medium prepared from L cells expressing wnt3a can replace feeder cell layers and medium containing LIF in maintaining ES cells in the proliferation without differentiation (self-renewal) state. By contrast, conditioned medium from NIH3T3 cells expressing wnt11 did not. Alkaline phosphatase staining and compact colony formation were used as criteria of cells being in the undifferentiated state. ES cells maintained in medium conditioned by Wnt3a expressing cells underwent freezing and thawing while maintaining properties seen with LIF maintained ES cells. Purified wnt3a did not maintain self-renewal of ES cells for prolonged intervals. Thus, other factors in the medium conditioned by wnt3a expressing cells may have contributed to maintenance of ES cells in a self-renewal state. Pluripotency of ES cells was determined with the use of embryoid bodies in vitro. PD98059, a MEK specific inhibitor, promoted the growth of undifferentiated ES cells maintained in conditioned medium from wnt3a expressing cells. By contrast, the P38 MAPK inhibitor SB230580 did not, suggesting a role for the MEK pathway in self-renewal and differentiation of ES cells maintained in the wnt3a cell conditioned medium. Thus, our results show that conditioned medium from wnt3a but not wnt11 expressing cells can maintain ES cells in self-renewal and in a pluripotent state. PMID- 16707110 TI - Protein kinase Calpha regulates insulin receptor signaling in skeletal muscle. AB - Certain PKC isoforms are stimulated by insulin and interact with IR as well as with IRS, but it is still not clear if specific PKC isoforms regulate IR signaling directly or through IRS-1. PKCalpha may regulate IRS activity in response to insulin. We investigated the possibility that PKCalpha may be important in insulin signaling. Studies were conducted on skeletal muscle in adult mice and on L6 skeletal cells. PKCalpha is constitutively associated with IRS-1, and insulin stimulation of PKCalpha causes disassociation of the two proteins within 5 min. Blockade of PKCalpha inhibited insulin-induced disassociation of PKCalpha from IRS1. Selective inhibition of PKCalpha increased the ability of insulin to reduce blood glucose levels. Insulin stimulation activates PKB and increases the association of PKCalpha with PKB. Blockade of PKCalpha increased threonine phosphorylation of PKB. We suggest that PKCalpha regulates insulin signaling in skeletal muscle through its disassociation from IRS-1 and association with PKB. PMID- 16707111 TI - The Rep protein binding elements of the plasmid ColE2-P9 replication origin. AB - The plasmid ColE2-P9 (ColE2) origin (32bp) is specifically recognized by the plasmid-specified Rep protein that initiates DNA replication. The ColE2 origin is divided into at least three functional subregions (I, II, and III), and three sites (a, b, and c) found in subregions I and II play important roles in Rep protein binding. We performed SELEX experiments of plasmid ColE2 to determine the optimal sequences for specific binding of the Rep protein. From these experiments, we obtained a common 16-bp sequence (5'-TGAGACCANATAAGCC-3'), which corresponds to about one half of the minimal ColE2 origin and contains sites a and b. Gel mobility shift assays using single-point mutant origins and the Rep protein further indicated that high affinity sequence-specific recognition by the Rep protein requires sites a, b, and c, but that mutations in site c were less disruptive to this recognition than those in sites a and b. PMID- 16707112 TI - Femtosecond dynamics of proteoheparan sulfate (HS-PG) after UV excitation--a readout for arteriosclerotic nanoplaque formation? AB - Ultrashort UV laser pulses were used to excite tryptophan residues of heparan sulfate proteoglycan (HS-PG) in blood substitute Krebs solution. Tryptophan fluorescence is sensitive to the environment, so its shift and decay indicate the conformation and solvation state of the protein. We monitored stimulated emission and excited-state absorption by probing with delayed white-light femtosecond pulses. Comparison with bare tryptophan revealed transient absorption features which are characteristic for HS-PG. Furthermore, the effect of adding calcium salt was investigated. Differences in the spectra from solutions with and without calcium developed during several minutes, which points to changes in protein conformation, but could only be measured in the sub-ps regime. These results provide a first step to a better understanding of the molecular formation of nanoplaques in blood vessels. The goal of this work is to open a way towards biosensing of the initial stages in atherogenesis allowing for a risk assessment in cardiovascular disease. PMID- 16707113 TI - STAT3-independent inhibition of lysophosphatidic acid-mediated upregulation of connective tissue growth factor (CTGF) by cucurbitacin I. AB - Cucurbitacins are recognised as anti-tumour agents because of their interference with STAT3 signalling, but may also affect the integrity of the actin cytoskeleton. In the present study the effect of cucurbitacin I was investigated in fibroblasts. In these cells, cucurbitacin I interfered with lysophosphatidic acid (LPA) signalling. It inhibited tyrosine phosphorylation of focal adhesion proteins and induction of connective tissue growth factor (CTGF), a potent profibrotic protein. Inhibition of Src family kinases with PP2, but not the inactive analogue PP3, also interfered with LPA-mediated tyrosine phosphorylation and induction of CTGF. Jak2-STAT3 signalling seemed to be the connecting link, because CTGF induction was sensitive to AG490, an inhibitor of Jak2, and cucurbitacin I, an inhibitor of Jak2 and STAT3. However, LPA did not activate tyrosine phosphorylation of STAT3. Furthermore, cucurbitacin I was as effective in STAT3 knock out cells as in control cells. Therefore, the inhibitory effect of cucurbitacin I was not related to inhibition of STAT3. Immunocytochemical analysis of cucurbitacin I-treated cells revealed disassembly of F-actin fibres, reorganisation into F-actin patches and resolution of focal adhesions. The phenotypic changes resembled changes observed after treatment of the cells with cytochalasin D, which has been shown to interfere with CTGF induction. Concentrations of cucurbitacin I, which have been shown to target Jak2-STAT3 signalling, thus, profoundly affect the actin cytoskeleton and may therefore modulate cell morphology, migration, adherence and gene expression also in non tumour cells. PMID- 16707114 TI - Nicotine modulates expression of amyloid precursor protein and amyloid precursor like protein 2 in mouse brain and in SH-SY5Y neuroblastoma cells. AB - Epidemiological studies indicate that tobacco smoking can be protective against neurodegenerative diseases such as Alzheimer's disease (AD) and Parkinson's disease (PD). The objective of the present study was to examine the changes in gene expression induced by chronic oral nicotine administration (100 mug/ml in 2% saccharin for 14 days), with special emphasis on amyloid precursor protein (APP) and its homologue, amyloid precursor-like protein 2 (APLP2), in different brain regions of C57BL/6 mice using a pathway-focused microarray. Our results revealed that nicotine stimulated mRNA expression of APP in the amygdala (64%; P = 0.003) and hippocampus (32%; P = 0.034) and of APLP2 in the amygdala (39%; P = 0.002). These results were verified by quantitative real-time RT-PCR except that expression of APLP2 was also significantly upregulated by nicotine in the hippocampus. In addition, in vitro nicotine treatment of SH-SY5Y neuroblastoma cells resulted in a significant increase in expression of APP protein, soluble APP, and APLP2, whereas co-treatment with mecamylamine (an antagonist of nicotinic acetylcholine receptors) attenuated the stimulating effect of nicotine on APP and APLP2 expression. These findings suggest that nicotine treatment facilitates the increase in the expression of mRNA and protein of the APP and APLP2 genes in rat brain and SH-SY5Y neuroblastoma cells. PMID- 16707115 TI - Distinct spatiotemporal expression of EFA6D, a guanine nucleotide exchange factor for ARF6, among the EFA6 family in mouse brain. AB - The EFA6 family is a member of guanine nucleotide exchange factors (GEFs) that can activate ARF6 specifically in vitro. In this study, we determined the complete primary sequence of mouse EFA6D encoding a protein of 1004 amino acids with a calculated molecular weight of 111,207 Da. In ARF pull-down assay, EFA6D showed a preferential GEF activity toward ARF6. RT-PCR analysis revealed the widespread tissue distribution of EFA6D and the high expression of EFA6A, C and D in the brain. In situ hybridization analysis demonstrated a distinct spatiotemporal expression pattern of EFA6D from those of EFA6A and C in mouse brain. Furthermore, immunoblot analysis revealed that EFA6D was highly concentrated in the postsynaptic density fraction. These findings suggest differential spatiotemporal regulation of ARF6 by three members of the EFA6 family in the brain. PMID- 16707116 TI - Autonomic and respiratory responses to microinjection of L-glutamate into the commissural subnucleus of the NTS in the working heart-brainstem preparation of the rat. AB - Changes in heart rate (HR), thoracic sympathetic nerve activity (tSNA) and frequency of phrenic nerve discharge (PND) in response to microinjection of L glutamate before and after local microinjection of ionotropic or metabotropic glutamate receptors antagonists into the commissural subnucleus of the NTS (comNTS) were investigated. The experiments were performed in an in situ unanesthetized decerebrated working heart-brainstem preparation (WHBP), and the main findings were as follows: (a) microinjection of increasing concentrations of L-glutamate (5, 25, 50, 250 and 500 mM) into the comNTS produced bradycardia, increase in tSNA and reduction in the frequency of the PND in a concentration dependent manner; (b) both bradycardia and increase in tSNA were almost abolished by kynurenic acid (KYN, 250 mM, a nonselective ionotropic glutamate receptor antagonist); (c) the reduction in the frequency of the PND was reversed to an increase in the frequency of the PND after KYN and this increase was blocked by the sequential microinjection of MCPG (100 mM, a nonselective metabotropic glutamate receptor antagonist); and (d) microinjection of increasing concentrations of trans-ACPD (0.5, 1.0, 2.5, 5.0 and 10 mM, a metabotropic glutamate receptor agonist), elicited bradycardia and increase in the frequency of the PND in a concentration-dependent manner, which were blocked by MCPG. Taken together, these data indicate that l-glutamate and its ionotropic receptors are involved in the sympathoexcitatory, bradycardic and reduction in the frequency of the PND responses whereas/although its metabotropic receptors are involved in the bradycardic and mainly in the increase in the frequency of the PND to microinjection of L-glutamate into the comNTS in the WHBP. PMID- 16707117 TI - An isocratic high performance liquid chromatographic method for quantification of mycophenolic acid and its glucuronide metabolite in human serum using liquid liquid extraction: application to human pharmacokinetic studies. AB - BACKGROUND: Mycophenolate mofetil (MMF) is a morpholinoethyl ester of mycophenolic acid (MPA), which is widely used as an immunosuppressive agent in renal transplant patients for the prophylaxis of acute rejection. We describe a fast and sensitive isocratic HPLC method for simultaneous quantification of the immunosuppressant mycophenolic acid, and its major metabolites in human serum. METHODS: The analytes were extracted from serum using ethyl acetate/2 propanol (4 1, v/v) and subjected to an isocratic HPLC method using a phenyl analytical column. A mobile phase consisted of methanol-0.05 mol/l sodium phosphate buffer (46/54 v/v; pH 2.5) containing hexadecyl trimethylammonium bromide (100mg/l w/v) and triethylamine (0.25% v/v) was used. RESULTS: The standard curve was linear from 0.050 to 51.2 microg/ml and 0.125 to 64 microg/ml, for mycophenolic acid and its metabolite, respectively. The method showed excellent selectivity, specificity, sensitivity, precision and accuracy. The respective limits of quantification for the drug and its metabolite were 0.050 and 0.125 microg/ml. This method was applied in a bioequivalence study following single dose administration of 2 different mycophenolate mofetil preparations in 24 healthy volunteers. Blood samples were analyzed and pharmacokinetic parameters of mycophenolic acid and its metabolite were compared. CONCLUSION: This procedure is simple, and comparing to the previously published methods, more sensitivity is obtained and less time is needed for sample preparation. Less time was needed for the liquid-liquid extraction. Although, suitability of the method has been demonstrated in pharmacokinetic studies of MMF in normal subjects, its specificity in renal and hepatic transplant patients has not been established and the stated upper linearity of MPAG may not be sufficient for use in transplant patients. PMID- 16707118 TI - Simultaneous determination of 12 steroids by isotope dilution liquid chromatography-photospray ionization tandem mass spectrometry. AB - BACKGROUND: Serum steroid assays play an important role in the clinical evaluation of a number of common endocrine disorders. Among various assays, tandem mass spectrometry (MS/MS) has being increasingly applied in clinical laboratories for its high sensitivity, specificity, and simultaneous multi analyte quantitation capability. Our first generation isotope dilution steroid profile assay by HPLC-tandem MS/MS with a C-18 column allowed for the measurement of 9 steroids in 18 min employing a sample volume of 760 ul serum. We describe our second generation steroid profile assay which allows for the quantitation of 12 steroids simultaneously employing HPLC-MS/MS and isotope dilution tandem MS in 11 min. This method requires a sample volume of 200 microl. METHODS: An API-5000 triple-quadrupole mass spectrometer (Sciex, Concord, Canada) coupled with the PhotoSpray source and Shimadzu HPLC system (Shimadzu Scientific Instruments, Columbia, MD) was used employing isotope dilution with deuterium labeled internal standard (IS) for each analyte. Two hundred microliters of serum were deproteinized by adding 300 microl of acetonitrile containing internal standards. After centrifugation, 450 microl of supernatant were diluted with 900 microl of water and 1000 microl aliquot were injected onto a C-8 column. After a 3 min wash the valve was activated to initiate the gradient elution program which eluted the steroids. Quantitation by MRM analysis was performed both in positive ion mode for 11 analytes and in negative ion mode for aldosterone. Within-day and between day precision, reliability and accuracy of this method were assessed by correlation with other MS/MS and immunoassay methods and by recovery study. RESULTS: Within-day CVs were <11.5% for all analytes tested and between-day CVs ranged from 3.5% to 12.2%. The results of the comparison study yield r values ranging between 0.908 and 0.999. Recovery ranged from 90% to 110%. CONCLUSIONS: This method can simultaneously measure 12 steroids in serum within 11 min with minimal sample preparation. It can be routinely employed in a clinical environment and is attractive because of its simplicity in sample processing and high throughput. PMID- 16707119 TI - Infant-directed speech supports phonetic category learning in English and Japanese. AB - Across the first year of life, infants show decreased sensitivity to phonetic differences not used in the native language [Werker, J. F., & Tees, R. C. (1984). Cross-language speech perception: evidence for perceptual reorganization during the first year of life. Infant Behaviour and Development, 7, 49-63]. In an artificial language learning manipulation, Maye, Werker, and Gerken [Maye, J., Werker, J. F., & Gerken, L. (2002). Infant sensitivity to distributional information can affect phonetic discrimination. Cognition, 82(3), B101-B111] found that infants change their speech sound categories as a function of the distributional properties of the input. For such a distributional learning mechanism to be functional, however, it is essential that the input speech contain distributional cues to support such perceptual learning. To test this, we recorded Japanese and English mothers teaching words to their infants. Acoustic analyses revealed language-specific differences in the distributions of the cues used by mothers (or cues present in the input) to distinguish the vowels. The robust availability of these cues in maternal speech adds support to the hypothesis that distributional learning is an important mechanism whereby infants establish native language phonetic categories. PMID- 16707120 TI - Evidentiality in language and cognition. AB - What is the relation between language and thought? Specifically, how do linguistic and conceptual representations make contact during language learning? This paper addresses these questions by investigating the acquisition of evidentiality (the linguistic encoding of information source) and its relation to children's evidential reasoning. Previous studies have hypothesized that the acquisition of evidentiality is complicated by the subtleness and abstractness of the underlying concepts; other studies have suggested that learning a language which systematically (e.g. grammatically) marks evidential categories might serve as a pacesetter for early reasoning about sources of information. We conducted experimental studies with children learning Korean (a language with evidential morphology) and English (a language without grammaticalized evidentiality) in order to test these hypotheses. Our experiments compared 3- and 4-year-old Korean children's knowledge of the semantics and discourse functions of evidential morphemes to their (non-linguistic) ability to recognize and report different types of evidential sources. They also compared Korean children's source monitoring abilities to the source monitoring abilities of English-speaking children of the same age. We found that Korean-speaking children have considerable success in producing evidential morphology but their comprehension of such morphology is very fragile. Nevertheless, young Korean speakers are able to reason successfully about sources of information in non-linguistic tasks; furthermore, their performance in these tasks is similar to that of English speaking peers. These results support the conclusion that the acquisition of evidential expressions poses considerable problems for learners; however, these problems are not (necessarily) conceptual in nature. Our data also suggest that, contrary to relativistic expectations, children's ability to reason about sources of information proceeds along similar lines in diverse language-learning populations and is not tied to the acquisition of the linguistic markers of evidentiality in the exposure language. We discuss implications of our findings for the relationship between linguistic and conceptual representations during development. PMID- 16707122 TI - Enhanced neuropeptide Y immunoreactivity and vasoconstriction in mesenteric small arteries from the early non-obese diabetic mouse. AB - The present study investigated whether sympathetic neurotransmission is altered at an early stage of diabetes in mesenteric small arteries isolated from female non-obese diabetic (NOD) and control animals without diabetes from the same mouse strain. The NOD diabetic mice had increased plasma glucose and hypertension. Confocal microscopy showed distribution of nerve terminals was similar, but immunoreaction intensity for neuropeptide Y (NPY) and tyrosine hydroxylase was higher in small arteries from NOD diabetic compared with NOD control mice. In the presence of prazosin and activated with vasopressin, electrical field stimulation evoked contractions which were more pronounced in mesenteric arteries from NOD diabetic versus NOD control mice and inhibited by the NPY Y(1) receptor antagonist, BIBP 3226. NPY concentration-response curves were leftward shifted in arteries from NOD diabetic versus NOD control both in arteries with and without endothelium, but not in the presence of the BIBP 3226. The present findings suggest that enhanced NPY content and vasoconstriction to NPY by activation of NPY Y(1) receptors in arteries from diabetic mice may contribute to the enhanced sympathetic nerve activity and vascular resistance in female non-obese early diabetic animals. PMID- 16707121 TI - Functional compensation in Hedgehog signaling during mouse prostate development. AB - Studies of hedgehog signaling in prostate development using anti-Shh antibodies, chemical inhibitors of hedgehog signaling and Shh(-/-) mutant mice have yielded conflicting data regarding the requirements of hedgehog signaling for normal ductal budding and glandular morphogenesis. We used transgenic mouse models in combination with chemical inhibitors and renal grafting to clarify the role of Hh signaling in prostate development. These studies showed that genetic loss of Shh is accompanied by an up-regulation of Indian Hedgehog (Ihh) and maintenance of Hh pathway activity. We found that while neither Gli1 nor Gli3 are required for normal prostate ductal budding, the urogenital sinus (UGS) of the Gli2(-/-) mutant mouse displays aberrant ductal budding in utero. When grown as a subcapsular graft, the Gli2(-/-) UGS exhibited prostatic differentiation but also displayed areas of focal epithelial hyperplasia. Functional redundancy between the three Gli transcription factors appears to mitigate the effect of Gli2 LOF as evidenced by residual Hh pathway activity in the E14 Gli2(-/-) UGS that could be inhibited by cyclopamine treatment. Together, these studies reveal a surprising degree of functional redundancy operating both at the level of the ligand and at the level of transcriptional regulation that effectively mitigates phenotypes associated with Hh-signaling perturbations. PMID- 16707123 TI - Involvement of the N-terminal unique domain of Chk tyrosine kinase in Chk-induced tyrosine phosphorylation in the nucleus. AB - Chk tyrosine kinase phosphorylates Src-family kinases and suppresses their kinase activity. We recently showed that Chk localizes to the nucleus as well as the cytoplasm and inhibits cell proliferation. In this study, we explored the role of the N-terminal unique domain of Chk in nuclear localization and Chk-induced tyrosine phosphorylation in the nucleus. In situ binding experiments showed that the N-terminal domain of Chk was associated with the nucleus and the nuclear matrix. The presence of the N-terminal domain of Chk led to a fourfold increase in cell population exhibiting Chk-induced tyrosine phosphorylation in the nucleus. Expression of Chk but not kinase-deficient Chk induced tyrosine phosphorylation of a variety of proteins ranging from 23 kDa to approximately 200 kDa, especially in Triton X-100-insoluble fraction that included chromatin and the nuclear matrix. Intriguingly, in situ subnuclear fractionations revealed that Chk induced tyrosine phosphorylation of proteins that were associated with the nuclear matrix. These results suggest that various unidentified substrates of Chk, besides Src-family kinases, may be present in the nucleus. Thus, our findings indicate that the importance of the N-terminal domain to Chk-induced tyrosine phosphorylation in the nucleus, implicating that these nuclear tyrosine phosphorylated proteins may contribute to inhibition of cell proliferation. PMID- 16707124 TI - A rat model of glaucoma induced by episcleral vein ligation. AB - To establish a reliable animal model of glaucoma, we examined if episcleral vein ligation in rat eyes can induce intraocular pressure (IOP) elevation and concomitant characteristic morphological features of glaucoma. IOP elevation was detected on the next day (30.1+/-4.4 mmHg: operated eyes; 21.0+/-1.8 mmHg: control eyes) and persisted at least 7 months after the procedure (24.5+/-2.3 mmHg: operated eyes; 19.7+/-1.9 mmHg: control eyes). These results suggest that episcleral vein ligation can induce very mild IOP elevation immediately after the operation, which can last over several months. Furthermore, it appears there was little variability in the patterns of IOP elevation among the individual eyes treated with episcleral vein ligation. Morphological changes were detected selectively in the retinal ganglion cell (RGC) layer and optic disc excavation was evident in the late stage of chronic IOP elevation. RGCs were selectively lost by apoptotic death. The number of RGCs was reduced by 18% at 12 weeks and eventually by 35% at 8 months postoperatively. Muller cells downregulated the expression of p27Kip1 and appeared to be partially in a reactive state even at the advanced stages of glaucoma. The expression of basic fibroblast growth factor and ciliary neurotrophic factor, which are neurotrophic factors implicated in the control of cell survivals and neuroprotection, significantly declined at the advanced stages. Taken altogether, these observations indicate that the episcleral vein ligation model based on the simple ligation procedure reproducibly provides a reliable glaucoma model and contributes to give insights into the underlying molecular and cellular bases of human glaucoma and to devise the new medication upon the disease. PMID- 16707125 TI - Toxoplasma gondii: an evaluation of diagnostic value of recombinant antigens in a murine model. AB - Laboratory diagnostics of toxoplasmosis depends primarily on serological methods detecting specific antibodies. Since these methods do not always enable specific and sensitive recognition of the infection and phase of toxoplasmosis, the search for new diagnostic tools continues. Recombinant antigens promise a new alternative in diagnostics of Toxoplasma gondii infections. In this work the usefulness of six recombinant T. gondii antigens: GRA1, GRA6, GRA7, p35, SAG1, and SAG2 in the detection of primary murine toxoplasmosis was evaluated. Sera obtained from infected mice differing in their natural susceptibility to T. gondii infection, BALB/c (relatively resistant) and C57BL/6 (relatively susceptible), were tested using ELISA. During acute infection high response to GRA7, GRA6, and p35 antigens was noticed, whereas a strong reactivity with surface antigens SAG1 and SAG2 was characteristic for chronic toxoplasmosis. Our results show that the recombinant antigens are useful in distinguishing between acute and chronic toxoplasmosis regardless of the genetically determined susceptibility of the host. PMID- 16707127 TI - Leishmania donovani: intracellular ATP level regulates apoptosis-like death in luteolin induced dyskinetoplastid cells. AB - Leishmaniasis presents a spectrum of diseases ranging from benign cutaneous lesions to the often-fatal visceralizing form. Luteolin, a dietary flavone induces apoptosis-like death in both promastigote and amastigote forms of Leishmania, the causative agent of the diseases. Here, we have elucidated the mechanism of action of luteolin by analyzing the mitochondrial and cytosolic changes associated with apoptosis-like death of leishmanial cells. In Leishmania donovani, treatment with luteolin induces the loss of both maxicircles and minicircles which resulted in the formation of dyskinetoplastid cells. The loss of mitochondrial DNA causes reduction in the activities of complex I, II, III, and IV of electron transport chain. However, the mitochondrial ATPase activity of complex V remains almost unaltered during treatment with luteolin but the sensitivity to oligomycin is lost. The inactivation of ETC complex is associated with decrease in mitochondrial as well as glycolytic ATP production, which is responsible for depolarization of Deltapsi(m) and alteration in mitochondrial structure. This event is followed by the release of cytochrome c from mitochondria in mt-DNA depleted leishmanial cells and causes an activation of caspase like proteases. Collectively our results provide the first insight into the mechanistic pathway of apoptosis-like death where inhibition of glycolytic ATP production is an essential event responsible for depolarization of Deltapsi(m) in mt-DNA depleted cells to propagate apoptosis-like death in leishmanial cells. PMID- 16707126 TI - The evolution of the new permeability pathways in Plasmodium falciparum--infected erythrocytes--a kinetic analysis. AB - Malaria parasites demonstrably increase the permeability of the membrane of the erythrocyte in which they develop and propagate. New permeability pathways (NPPs) generated by parasite activity and identified in the erythrocyte membrane are held responsible for these changes. Here, we present a novel analysis of hemolysis curves of infected cells in iso-osmotic solutions of solutes that penetrate selectively into infected cells, as a function of parasite development. The analysis yields three parameters: the t(1/2) of lysis (reciprocally related to permeability), the maximal lysis, and a parameter that expresses the variation of the cell population. Different developmental stages of the parasite were obtained either by sampling synchronized cultures with time or by the fractionation of asynchronous cultures on a Percoll-sorbitol density gradient. While the results confirm previous reports on the stage-dependent evolution of NPPs, they also reveal that the evolution of NPPs is not synchronous: NPPs evolve differentially throughout the ring stage and only at the mid-trophozoite stage they are fully deployed in the majority of the infected cells, but not in all. This leads to desynchronization in the culture and to less than the maximal possible rate of multiplication. PMID- 16707128 TI - Differentiation of monocyte-derived dendritic cells is associated with upregulation and activation of Rac-1 small GTPase. AB - Critical changes occurring in Rac-1 molecule, a cytoskeleton organizing small GTPase associated with cell ruffling, have been analyzed in dendritic cells (DCs) derived from monocytes cultured with granulocyte-macrophage colony-stimulating factor and IFN-alpha or IL-4. Although with different kinetics, both agents induced activation of Rac-1 molecule and, more importantly, an upregulation of both protein expression and mRNA transcription. These findings strengthen the role of Rac-1 molecule in the induction of DC differentiation and suggest that, besides its activation, the upregulation of Rac-1 molecule might also play a role in the acquisition of DC mature phenotype. PMID- 16707129 TI - Molecular cloning, recombinant expression, and growth-promoting effect of mud carp (Cirrhinus molitorella) insulin-like growth factor-I. AB - A full-length cDNA encoding insulin-like growth factor-I (IGF-I) was cloned from mud carp (Cirrhinus molitorella) liver tissue using reverse transcription polymerase-chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) techniques. The IGF-I precursor cDNA consists of 822 bp in size with a 218 bp 5' untranslated region and 118 bp 3'-untranslated region. The 486 bp open reading frame encodes a 161 amino acid peptide with a molecular weight of 17.9 kDa. The deduced IGF-I amino acid sequence shared 82.5-97% and 82.5-84% sequence identity with fish and mammalian counterparts, respectively. The mature IGF-I was overexpressed in Escherichia coli, and the expression level of recombinant mcIGF I reached to 34.1% of the cell total protein. After purification and refolding of recombinant mcIGF-I, growth-promoting effect of recombinant mcIGF-I was investigated, the results showed that the recombinant mcIGF-I significantly enhanced the growth rate of juvenile tilapia. After 6-week treatment, the growth rates of group 1 and 2 were 53 and 67.3% higher than the saline-treated control group. The recombinant mcIGF-I was more effective than recombinant mcGH to enhance the growth rate of juvenile tilapia. The recombinant mcIGF-I-treated fish revealed no significant changes of content of protein, lipid, ash and moisture in muscle. PMID- 16707130 TI - Determination of nitrogen mustard hydrolysis products, ethanolamines by gas chromatography-mass spectrometry after tert-butyldimethylsilyl derivatization. AB - A method for determining N-ethyldiethanolamine (EDEA), N-methyldiethanolamine (MDEA) and triethanolamine (TEA), hydrolysis products of nitrogen mustards, in water, urine and blood samples using gas chromatography-mass spectrometry (GC-MS) after derivatization by tert-butyldimethylsilylation (TBDMS) is described. The sample solution was evaporated to dryness, and reacted with N-methyl-N-(tert butyldimethylsilyl)trifluoroacetamide (MTBSTFA) at 60 degrees C for 1h. The TBDMS derivatives were separated on a DB-5 column and detected by electron-ionization MS. The quantitation of EDEA, MDEA and TEA was performed by measuring the respective peak areas on the extracted ion chromatograms of m/z 216, m/z 202 and m/z 346, respectively, using nonadecane (C19), the peak area of which was measured at m/z 268, as an internal standard. When the water sample was initially analyzed, considerable loss of EDEA, MDEA and TEA occurred by evaporation. The addition of hydrochloric acid (HCl) to the water sample (final 1 mM), however, permitted quantitative recoveries to be achieved (88%, 88% and 79% for EDEA (TBDMS)2, MDEA-(TBDMS)2 and TEA-(TBDMS)3, respectively). The limits of detections (LODs, scan mode, S/N = 3) were 2.5, 2.5 and 10 ng/ml for EDEA, MDEA and TEA, respectively. Ethanolamines could be also determined in urine samples (volume 0.1 ml), with reasonable recoveries of 72-100% by the addition of HCl (final 1 mM). For the analysis of serum samples, the sample was precipitated by the addition of perchloric acid (final 3.2%), and the resulting supernatant was neutralized with potassium carbonate, and then acidified by the addition of HCl. The recovery of TBDMS derivatives of ethanolamines was found to rather low (7-31%). PMID- 16707131 TI - Synthesis of flexible silica aerogels using methyltrimethoxysilane (MTMS) precursor. AB - The experimental results on the synthesis of flexible and superhydrophobic silica aerogels using methyltrimethoxysilane (MTMS) precursor by a two-step (acid-base) sol-gel process followed by the supercritical drying, are reported. The effects of various sol-gel parameters on the flexibility of the aerogels have been investigated. The aerogels of different densities were obtained by varying the molar ratio of MeOH/MTMS (S) from 14 to 35, with lower densities for larger S values. It has been observed that the Young's modulus (Y) decreased from 14.11 x 10(4) to 3.43 x 10(4) N/m(2) with the decrease in the density of the aerogels from 100 to 40 kg/m(3). Simultaneously, the aerogels are superhydrophobic with a contact angle as high as 164 degrees . The superhydrophobic aerogels are thermally stable up to a temperature of 530 K, above which they become hydrophilic. The aerogels have been characterized by bulk density, percentage volume shrinkage, and porosity measurements. The microstructures of the aerogels have been studied using the transmission electron microscopy (TEM). The Young's modulus of the aerogels has been determined by an uniaxial compression test. The variation of physical properties of the aerogels has been explained by taking into consideration the hydrolysis, condensation reactions, the resulting colloidal clusters and their network formation. PMID- 16707132 TI - Photocatalytic degradation of aniline at the interface of TiO2 suspensions containing carbonate ions. AB - The degradation of aniline has been investigated using aqueous TiO2 suspensions containing carbonate ions as photocatalyst. The addition of carbonate to Degussa P-25 increased the number of active adsorption sites at its surface. For the TiO2 suspensions containing carbonate ions the intensity of adsorption of aniline increased to 6.9 x 10(2) from 5.5 x 10(2) mol(-1) dm(3) in case of bare TiO2 suspensions. This in turn results in the increased interfacial interaction of the photogenerated charge carriers with the adsorbed aniline and thus enhancing the rate of its photodecomposition to 6.5 x 10(-6) mol dm(-3) s(-1) compared to 2.7 x 10(-6) mol dm(-3) s(-1) in the absence of Na(2)CO(3). The maximum efficiency of this photocatalyst has been obtained upon addition of 0.11 mol dm(-3) of Na(2)CO(3) at pH 10.8. The photocatalytic action is understood by the simultaneous interaction of intermediates, *OH and CO*-(3), and their reactivity with aniline. Azobenzene, p-benzoquinone, nitrobenzene, and NH(3) have been identified as the major products of the photooxidation of aniline. Both the reactant and products have been followed kinetically. The photodegradation follows Langmuir-Hinshelwood Model. The mechanism of the occurring reactions has been analyzed and discussed. In the presence of Na(2)CO(3), 3 x 10(-3) mol dm(-3) of aniline could be photodegraded completely in about 6 h while all organic intermediates decomposed completely within about 10 h. PMID- 16707133 TI - Wet-chemical synthesis of crystalline BaTiO3 from stable chelated titanium complex: formation mechanism and dispersibility in organic solvents. AB - Crystalline barium titanate nanoparticles were synthesized in solution at low temperature (70 degrees C) from acetylacetone chelated titanium complex and barium hydroxide. Very fine crystalline solids were characterized to cubic phase of BaTiO(3) by X-ray diffraction studies of the air-dried samples. It was observed that the crystalline barium titanate was formed in solution at Ba/Ti molar ratio > or =2.5. The dependence of the reaction temperature and the Ba(OH)(2) concentration (in terms of Ba/Ti molar ratio) on formation of crystalline BaTiO(3) in solution-phase was studied, and a plausible mechanism toward the formation of crystalline BaTiO(3) was also proposed. Crystallite sizes of the BaTiO(3) were found to be in the range 33-50 nm, while the average particle sizes, measured by dynamic light scattering method were in the range 70 100 nm. The crystalline BaTiO(3) prepared from acetylacetone chelated titanium complex was highly dispersible in organic medium such as N-methyl-2-pyrillidone (NMP) and N,N-dimethyl formamide (DMF). PMID- 16707134 TI - Effects of heavy metals and oxalate on the zeta potential of magnetite. AB - Zeta potential is a function of surface coverage by charged species at a given pH, and it is theoretically determined by the activity of the species in solution. The zeta potentials of particles occurring in soils, such as clay and iron oxide minerals, directly affect the efficiency of the electrokinetic soil remediation. In this study, zeta potential of natural magnetite was studied by conducting electrophoretic mobility measurements in single and binary solution systems. It was shown that adsorption of charged species of Co(2+), Ni(2+), Cu(2+), Zn(2+), Pb(2+), and Cd(2+) and precipitation of their hydroxides at the mineral surface are dominant processes in the charging of the surface in high alkaline suspensions. Taking Pb(2+) as an example, three different mechanisms were proposed for its effect on the surface charge: if pH<5, competitive adsorption with H(3)O(+); if 56, precipitation of heavy metal hydroxides prevails. Oxalate anion changed the associated surface charge by neutralizing surface positive charges by complexing with iron at the surface, and ultimately reversed the surface to a negative zeta potential. Therefore the adsorption ability of heavy metal ions ultimately changed in the presence of oxalate ion. The changes in the zeta potentials of the magnetite suspensions with solution pH before and after adsorption were utilized to estimate the adsorption ability of heavy metal ions. The mechanisms for heavy metals and oxalate adsorption on magnetite were discussed in the view of the experimental results and published data. PMID- 16707135 TI - Dry gangrene of the extremities in calves associated with Salmonella dublin infection; a possible immune-mediated reaction. AB - Dry gangrene of the extremities in calves is a circulatory error that may occur after infection with Salmonella dublin. This report describes an examination of three affected, 12 in-contact and five control calves, a main objective being to investigate the possible role of cold agglutination in pathogenesis. The lesions included dry gangrene of the hind legs, ears and tail. A cold agglutination test gave positive results in all animals examined except the controls. The three affected calves had high titres of S. dublin antibodies, as also did four of the in-contact animals. The results suggested a relationship between cold agglutination and the occurrence of the disease. PMID- 16707136 TI - Starvation period and age affect the response of female Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) to odor and visual cues. AB - The effects of starvation or age on the walking or flying response of female Frankliniella occidentalis to visual and/or odor cues in two types of olfactometer were examined in the laboratory. The response of walking thrips starved for 0, 1, 4, or 24h to an odor cue (1microl of 10% p-anisaldehyde) was examined in a Y-tube olfactometer. The take-off and landing response of thrips (unknown age) starved for 0, 1, 4, 24, 48 or 72h, or of thrips of different ages (2-3 days or 10-13 days post-adult emergence) starved for 24h, to a visual cue (98 cm(2) yellow sticky trap) and/or an odor cue (0.5 or 1.0 ml p-anisaldehyde) was examined in a wind tunnel. More thrips walked up the odor-laden arm in the Y tube when starved for at least 4h (76%) than satiated thrips (58.7%) or those starved for 1h (62.7%, P<0.05). In the wind tunnel experiments the percentage of thrips to fly or land on the sticky trap increased between satiated thrips (7.3% to fly, 3.3% on trap) and those starved for 4h (81.2% to fly, 29% on trap) and decreased between thrips starved for 48 (74.5% to fly, 23% on trap) and 72 h (56.5% to fly, 15.5% on trap, P<0.05). Fewer younger thrips (38.8%) landed on a sticky trap containing a yellow visual cue of, those that flew, than older thrips (70.4%, P<0.05), although a similar percentage of thrips flew regardless of age or type of cue present in the wind tunnel (average 44%, P>0.05). PMID- 16707137 TI - Multiple coding sequences for the genome-linked virus protein (VPg) in dicistroviruses. AB - N-terminal Edman sequencing of the genome-linked viral protein (VPg) of Plautia stali intestine virus (PSIV, Dicistroviridae) detected heterologus residues. The VPg sequence determined was found to be triplicated in the nonstructural protein precursor. Multiple VPg-like sequences were also found in 10 of the 12 dicistroviruses with a maximum of six copies in Solenopsis invicta virus-1. We postulate that redundant VPg coding sequences facilitate multiplication of dicistroviruses, because fewer cycle of translation of the nonstructural protein precursor produces larger amounts of VPg proteins in parallel with the increased production of capsid proteins by the intergenic internal ribosome entry site mediated translation. PMID- 16707138 TI - Susceptibility of the peachtree borer, Synanthedon exitiosa, to Steinernema carpocapsae and Steinernema riobrave in laboratory and field trials. AB - The nematode Steinernema carpocapsae (All) strain was significantly more effective against peachtree borer larvae (Synanthedon exitiosa [Lepidoptera: Sesiidae]) than Steinernema riobrave (7-12) strain in field and laboratory experiments. Eighty-eight percent control of peachtree borer larvae was obtained with S. carpocapsae in the field trial when applied at 3 x 10(5) infective juveniles per tree, and 92% mortality was obtained in the lab assay using 50 infective juveniles per larva. PMID- 16707139 TI - AMPD1 gene polymorphism and the vasodilatory response to ischemia. AB - Peripheral vasculature resistance can play an important role in affecting blood pressure and the development of cardiovascular disease. A better understanding of the genes that encode vasodilators, such as adenosine, will provide insight into the mechanisms underlying cardiovascular disease. We tested whether the adenosine monophosphate deaminase-1 (AMPD1) C34T gene polymorphism was associated with the vasodilatory response to ischemia in Caucasian females aged 18-35 years. Blood samples (n = 58) were analyzed for the C34T variant and resulted in the following genotype groups: CC (n = 45) and CT (n = 13). Mean blood pressure (MBP), heart rate, and forearm blood flow (FBF) measured by venous occlusion plethysmography were measured at baseline and at 1 (peak FBF), 2 and 3 min of vasodilation during reactive hyperemia following 5 min of arm ischemia. To control for interindividual variability in baseline FBF and forearm vascular resistance (FVR) the percent change in FBF and FVR were calculated for each min. The percent decrease in FVR was significantly greater in the CT compared to the CC genotype group (-40+/-4% vs. -24+/-3%, P = 0.01) during the 2nd min of reactive hyperemia. The percent increase in FBF tended to be greater in the CT compared to the CC genotype group (+69+/-9% vs. +42+/-9%, P = 0.07) during the 2nd min of reactive hyperemia after adjustment for percent body fat. Consistent with previous findings of increased production of adenosine during exercise in individuals carrying a T allele, our findings suggest that the AMPD1 C34T polymorphism is associated with vasodilatory response to ischemia in the peripheral vasculature because individuals with the T allele had a greater vasodilatory response to ischemia. PMID- 16707140 TI - Effect of repetitive ischemic preconditioning on spinal cord ischemia in a rabbit model. AB - A completely randomized controlled study based on a rabbit model was designed to study the effect of repetitive ischemic preconditioning (IPC) on a spinal cord ischemic reperfusion injury. Twenty four white adult Japanese rabbits were randomly assigned to one of the 3 groups (n = 8 per group): Group I: sham operation group, Group II: ischemic reperfusion group, and, Group III: IPC group. Spinal cord ischemia was induced by infra-renal aortic cross-clamp for 45 min in Group II. Before 45 min ischemia, the rabbits in Group III underwent four cycles of IPC (5 min of ischemia followed by 5 min of reperfusion). Post-operative neurological function, electromyography (EMG) of rear limbs, and spinal cord histopathological changes were measured. The concentrations of calcium, magnesium, copper, and zinc in spinal cord were measured in the 7th day. The neurological function and histopathological changes in Group II were significantly different from those in Group I or Group III (P < 0.05 or 0.01). There was a more significant change of EMG in Group II than that in Group III (P < 0.05). The concentrations of calcium and copper in Group II were significantly higher (P < 0.05 or 0.01), but magnesium and zinc were significantly lower (P < 0.05) than those in Group I. Calcium and copper in Group II were significantly higher (P < 0.05), but zinc was significantly lower (P < 0.01) than those in Group III. In conclusion, repetitive IPC can protect rabbit spinal cord from ischemic reperfusion injury in a timely manner, which is associated with corrections of imbalance of calcium, magnesium, copper, and zinc in the ischemic region. PMID- 16707141 TI - Caffeic acid phenethyl ester blocks free radical generation and 6-hydroxydopamine induced neurotoxicity. AB - Neurotoxicity induced by 6-hydroxydopamine (6-OHDA) is believed to be due, in part, to the production of reactive oxygen species (ROS). Antioxidants protect neurons against 6-OHDA-induced neurotoxicity by inhibiting free radical generation. In this study, we investigated whether or not caffeic acid phenethyl ester (CAPE) could protect neurons against 6-OHDA-induced neurotoxicity in cultured rat rostral mesencephalic neurons (RMN) and cerebellar granule neurons (CGN). We now report that exposure of RMN and CGN to 6-OHDA (40 microM for RMN and 70 microM for CGN) resulted in significant increases in free radical production and death of both neuron types. Pretreatment with CAPE (10 microM) for 2 h prevented both 6-OHDA-induced free radical generation and neurotoxicity. Furthermore, CAPE also attenuated H(2)O(2)-induced neurotoxicity. Our results strongly suggest that CAPE blocks 6-OHDA-induced neuronal death possibly by inhibiting 6-OHDA-induced free radical generation and blocking free radical induced neurotoxicity in neurons. Both the antioxidative and neuroprotective effects of CAPE may be beneficial in the therapy for Parkinson's disease and other neurodegenerative diseases. PMID- 16707142 TI - Sequential activation of caspases and synergistic beta-cell cytotoxicity by palmitate and anti-Fas antibodies. AB - To assess the mechanism of beta-cell lipotoxicity in comparison with Fas-mediated cell death, we used a mouse beta-cell clone stably transfected with human Fas. Palmitate induced beta-cell death in correlation with medium glucose levels between 5 and 20 mmol/l, while Fas-mediated cytotoxicity was observed irrespective of glucose concentration. At the glucose level of 10 mmol/l, palmitate induced caspase-6 activity within 3 h, and caspase-3 activity after a lag period of 6 h. The activities of caspases were correlated with glucose concentration. A caspase-6 inhibitor attenuated caspase-3 activation and cell death induced by palmitate. Oxfenicine, an inhibitor of carnitine palmitoyltransferase-1, attenuated both palmitate-induced cytotoxicity and activation of caspases. Finally, beta-cell cytotoxicity caused by the combination of anti-Fas and palmitate at 25 mmol/l of glucose was greater than the sum of those induced by each. These observations suggest that palmitate induces sequential activation of caspase-6 and caspase-3 through a mitochondrial signal(s), and caspase-6 plays a primary role in the mechanism. Fas-mediated beta cell death and lipotoxicity may share common mechanisms involving caspase activation, and thereby synergistically inducing beta-cell death, although upstream signaling pathways are distinct. PMID- 16707143 TI - Socio-economic status inequality and major causes of death in adults: a 5-year follow-up study in rural Vietnam. AB - OBJECTIVES: This study aimed to analyse the associations between cause-specific mortality in adults (aged 20 years and above) and socio-economic status (SES) in a rural setting of Vietnam during a time of economic transition. STUDY DESIGN: The study was carried out as part of the FilaBavi demographic surveillance system, with a dynamic cohort of 50,000 inhabitants from January 1999 to December 2003. METHODS: Causes of death in the adult population were derived using verbal autopsy. A Cox regression model was employed to check the association of SES with three major causes of death: communicable diseases; non-communicable diseases; and injuries. RESULTS: The crude mortality rates were 9.2 and 6.6 per 1000 person years in adult males and females, respectively. Men had higher mortality rates than women for all mortality categories and for all levels of education and household economic situation (HES). Mortality rates increased substantially with age, and showed similar age effects for all mortality categories with the strongest association for non-communicable diseases. Education was an important factor for survival in general, and high HES seemed to benefit men more than women. CONCLUSIONS: Interventions and policies to reduce exposure to risk factors for non-communicable diseases are needed in low-education groups. However, further study is needed to analyse the mortality inequity across all age groups. PMID- 16707144 TI - Risk factors for sporadic community-acquired Legionnaires' disease. A 3-year national case-control study. AB - OBJECTIVE: Risk factors for sporadic community-acquired Legionnaires' disease (LD) have been studied in the past, well before the widespread introduction of the urinary antigen test. Our objective was to evaluate the impact of the concomitant decrease in underdiagnosis on established and unknown risk factors for LD. STUDY DESIGN: Prospective case-control study. METHODS: From 1 July 1998 to 30 June 2001, 228 LD cases and 293 controls were included for a national case control study. Patients were included upon notification provided that they fulfilled international criteria for confirmed LD. RESULTS: A history of diabetes mellitus, current tobacco smoking, travelling abroad, spending one or more nights away from home not leaving the country and being a driver by profession were independent risk factors. LD patients who had travelled abroad during their incubation period differed from LD patients who had not. They appeared healthier than non- or domestic travellers with respect to a history of coronary disease, pulmonary disease, current use of corticosteroids or immunosuppressives and any medication. Also the environmental risk factors differed significantly for the two groups. CONCLUSIONS: The finding of two distinct populations of LD patients calls for a differentiated preventive strategy. PMID- 16707145 TI - The optics of vertebrate photoreceptors: anisotropy and form birefringence. AB - The optics of vertebrate photoreceptors have been investigated with specific reference to the effect of form birefringence. The complex dielectric tensor of the lamellar-like outer segment structure has been derived, allowing the transverse spectral absorbance to be calculated for different incident polarizations. These results were used to calculate the changes in the cellular dichroic ratio as a function of both the volume occupied by the bilayers and the real and complex parts of the intrinsic birefringence of the bilayers. Physiologically realistic values of these parameters show the cellular dichroic ratio to be greater than the bilayer dichroic ratio by a factor of approximately 1.3. Furthermore, the calculations of spectral absorbance indicate that form birefringence may affect measurements of optical density in transversely orientated outer segments. PMID- 16707146 TI - Dynamics of the influence of segmentation cues on orientation perception. AB - Contextual effects abound in vision. The tilt illusion (TI) is an example-a tilted surrounding annulus causes a vertical central pattern to appear rotated away from the surround. We investigate the dynamics of this effect by presenting components of the stimulus asynchronously. At equal contrast, the largest illusion occurs when centre and surround are presented simultaneously. We vary the spatial gap between centre and surround, the relative contrast and depth and find that these segmentation cues result in a reduced TI upon simultaneous presentation, but not all other times. This reveals the dynamics of orientation and other segmentation cue interactions. PMID- 16707147 TI - Polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and heavy metals (HMs) as well as their genotoxicity in soil after long-term wastewater irrigation. AB - A study was carried out on the residue level of major concern pollutants (PAHs, PCBs and HMs) and the assessment of their genotoxicity in soils obtained from Shenyang, northeast of China which had been subjected to wastewater irrigation for more than 40 years. Topsoils (0-20cm) in paddy fields were sampled along the upper, middle and lower reaches of the wastewater disposal channel. Sixteen USEPA PAHs were determined by High Performance Liquid Chromatography (HPLC) with fluorescence detector, eight PCBs were detected by Gas Chromatography (GC) with electron capture detector, and six heavy metals (Cd, Cu, Pb, Zn, Ni, and Cr) were measured by Atomic Absorption Spectrophotometer (AAS). The genotoxicity effect of soils was examined by Vicia faba micronucleus (MN) test. Archived soils that had undergone a similar history of wastewater irrigation provided by Technical University of Berlin, Germany were subjected to analysis of the above pollutants and Vicia faba/MN test for comparison. Results indicated elevated residues of the studied pollutants (PAHs, especially benzo (a) pyrene, the eight PCB congeners and heavy metals) in both tested and archived soils. The MN frequencies were 2.2 48.4 times higher compared with the control. However, there was no correlation between the MN frequencies and the concentration of pollutants detected. This investigation suggested a potential ecological risk even with a lower level of residual pollutants in soil matrix after long-term wastewater irrigation. PMID- 16707148 TI - Preparation of spherical iron nanoclusters in ethanol-water solution for nitrate removal. AB - In this study, a higher surface area spherical nanoscale zero valent iron (HNZVI) cluster (80 nm, 54.25 m(2)g(-1)) was synthesized in ethanol-water mixed solvent in the presence of dispersion agent of polyglycol (PEG). At the same time, a lower surface area nanoscale zero valent iron (LNZVI) particle (80 nm, 8.08 m(2)g(-1)) was also prepared with only de-ioned water as reaction media. Their structures, compositions and physical properties were characterized by transmission electron microscope (TEM), X-ray diffractometer (XRD), inductively coupled plasma atomic emission spectrophotometer (ICP-AES), and Brunauer-Emmett Teller (BET) surface area analyzer and the results obtained for these two kinds of nanoscale iron were compared with each other and also with those reported in the literatures. The HNZVI clusters seemed to be accumulated by smaller iron particles (<10 nm). At the same time, whiskers were formed in the final produce. Reactivity of the HNZVI was affirmed via denitrification of nitrate. The factors controlling the reduction of nitrate, such as pH, dissolved oxygen (DO), iron content as well as the initial nitrate concentration were also discussed. Finally, kinetic analysis revealed that chemical reduction of nitrate by HNZVI could not be described by the first- or pseudo-first-order kinetic model. PMID- 16707149 TI - Maintenance of a low-sodium, high-carotene and -vitamin C diet after a 1-year dietary intervention: the Hiraka dietary intervention follow-up study. AB - BACKGROUND: The importance of dietary modification for disease prevention is widely accepted. The difficulty of implementing and sustaining long-term changes is also well documented. Nevertheless, a few studies have attempted to achieve significant dietary change for extended periods. METHODS: The Hiraka Dietary Intervention Study was a community-based randomized cross-over trial designed to develop an effective dietary modification tool and system in an area with high mortality for stomach cancer and stroke in 1998-2000. The main study subjects were 550 healthy volunteers, who were randomized into two groups and given tailored dietary education aimed at decreasing the intake of sodium and increasing that of carotene and vitamin C in either the first or second year. Four (first intervention group) and three (second intervention group) years after the intervention ended, 308 subjects were selected for this follow-up dietary survey. RESULTS: The low-sodium, high-vitamin C and -carotene diet was maintained with only a small, nonsignificant reversal from post-intervention to follow-up (P = 0.082-0.824). Significant changes from pre-intervention to follow-up were also maintained (P < 0.01). CONCLUSION: This dietary intervention program was maintained well over 4 years after the termination of the intervention sessions. PMID- 16707150 TI - Basic and applied research on domestic, exotic and endangered carnivores. Proceedings of the 5th International Symposium on Canine and Feline Reproduction. August 4-6, 2004. Embu das Artes, Brazil. PMID- 16707151 TI - Biochemical effects of nonylphenol polyethoxylate adjuvant, Diquat herbicide and their mixture on the three-spined stickleback (Gasterosteus aculeatus L.). AB - This study examined the response of 7-ethoxyresorufine-O-deethylase, glutathione S-transferase, glutathione peroxidase, glutathione content, level of thiobarbituric acid reactive compounds and circulating vitellogenin, in three spined sticklebacks after 21 days of exposure to Diquat herbicide, commercial nonylphenol polyethoxylate adjuvant and mixture between Diquat and adjuvant. The results showed that adjuvant exerted more important oxidative effects than Diquat and that mixture effects were unlike to single additivity. This study argues for ecotoxicological risk assessment of adjuvants and mixtures of adjuvants and pesticides. PMID- 16707152 TI - Modeling of gene expression pattern alteration by p,p'-DDE and dieldrin in largemouth bass. AB - In this study, largemouth bass (LMB) were subchronically exposed to p,p'-DDE or dieldrin in their diet to evaluate the effect of exposure on expression of genes involved in reproduction and steroid homeostasis. Using real-time PCR, we detected a different gene expression pattern for each OCP, suggesting that they each affect LMB in a different way. We also detected a different expression pattern among sexes, suggesting that sexes are affected differently by OCPs perhaps reflecting the different adaptive responses of each sex to dysregulation caused by OCP exposure. PMID- 16707153 TI - Altered hepatic retinol and CYP26 levels in adult European common frogs (Rana temporaria) exposed to p,p'-DDE. AB - The effects of environmental contamination on amphibians are of particular concern because there are reports of declining numbers of species and individuals in most parts of the world during the last 50 years. During the last decade there has been increased focus on the role of persistent organic pollutants as retinoid (vitamin A) disrupters, and their effects on development, growth and sexual differentiation. To study the effects of p,p'-DDE, one of the most persistent metabolites of the pesticide DDT, on retinol homeostasis, we subcutaneously exposed adult male European common frogs (Rana temporaria) to different doses of p,p'-DDE (0, 0.01, 0.1, 1 and 10 mg/kg body mass) and studied the effect of a short term exposure (14 days) on hepatic retinoid levels and CYP26 gene and protein expression. Hepatic retinol concentrations, CYP26 gene and protein levels were analysed using HPLC, quantitative RT-PCR and indirect ELISA, respectively. Our results showed a significant p,p'-DDE dose-specific increase in the hepatic retinol concentration. CYP26 gene and protein expression were reduced in an apparent p,p'-DDE dose-specific manner. The results suggest that p,p'-DDE may interfere with the hepatic metabolism of retinol in adult frogs by decreasing CYP26 expression patterns. PMID- 16707154 TI - Effects of the model androgen methyltestosterone on vitellogenin in male and female eelpout, Zoarces viviparus (L). AB - In the present experiment male and female eelpout, Zoarces viviparus were exposed to different doses of the model androgen 17alpha-methyltestosterone (MT) and the effects on the plasma level of vitellogenin and the gonadosomatic index were investigated. In females exposed to different doses of MT (nominal concentrations 10, 25, 50, 100 and 500 ng/L) in the ambient seawater, the concentrations of the circulating yolk-precursor protein vitellogenin (vtg) were shown to decrease in all groups but only significantly in the MT-100 group when compared to controls. No significant effects could be observed on the GSI during early vitellogenesis (April/May). Males exposed to E2 under flow through conditions during 10 days showed an induced synthesis of vtg as depicted by a high level of circulating vtg. When the estrogen-exposed males were subsequently exposed to different doses of MT only, during another 10 days, the levels of vtg decreased significantly in three of the MT-treated groups when compared with the level of vtg in plasma of the estrogen-treated group after the first 10 days. The gonadosomatic index was observed to decrease by the estradiol-treatment but to increase in a dose dependent manner in the MT-exposed groups, indicating that MT had the capacity to override the effect of the preceding exposure to estradiol-17beta. PMID- 16707155 TI - Bioindicators in monitoring long term genotoxic impact of oil spill: Haven case study. AB - The evaluation of long term impact and risk of oil spill is a complex process involving chemical analyses and development of the ecosystem-based toxicology. An integrated biomarker approach using different bioindicators, mussels, oysters and fish with different feeding habits was applied to evaluate the long term risk from Haven oil spill along the Ligurian coast (Italy). Mussels were caged for a period of 4 weeks and fish were caught in the impacted and reference area. Caged oysters were also analyzed in different area of the wreck. DNA damage and micronuclei (MN) frequency were evaluated in gill cells of bivalves. DNA single strand breaks were measured in hepatocytes and MN were measured in fish erythrocytes. The results revealed an increase in MN frequency (more than 10 times the level at the reference site) in caged mussels from Arenzano compared to the reference area after an interval of 4 months from the accident. No increase in DNA damage and a significant increase in MN frequency were recorded in caged mussels (mean value 10.15 vs 5.3) and in benthic fish Mullus barbatus (2.5 vs 0.7) in a further sampling in 1998. Statistically significant increase of DNA damage and MN frequency was observed in caged oysters in different areas of the wreck in a biomonitoring carried out in 2001. PMID- 16707156 TI - Binding of anti-inflammatory drug cromolyn sodium to bovine serum albumin. AB - Fluorescence spectroscopy in combination with circular dichroism (CD) and UV-vis absorption spectroscopy were employed to investigate the binding of anti inflammatory drug cromolyn sodium (Intal) to bovine serum albumin (BSA) under the physiological conditions with Intal concentrations of 0-6.4 x 10(-5)mol L(-1). In the mechanism discussion, it was proved that the fluorescence quenching of BSA by Intal is a result of the formation of Intal-BSA complex. Quenching constants were determined using the Stern-Volmer equation to provide a measure of the binding affinity between Intal and BSA. The thermodynamic parameters Delta G, Delta H, Delta S at different temperatures (298, 304, and 310 K) were calculated and the results indicate the electrostatic interactions play a major role in Intal-BSA association. Binding studies concerning the number of binding sites (n=1) and apparent binding constant K(b) were performed by fluorescence quenching method. Utilizing fluorescence resonant energy transfer (FRET) the distance R between the donor (BSA) and acceptor (Intal) has been obtained. Furthermore, CD and synchronous fluorescence spectrum were used to investigate the structural change of BSA molecules with addition of Intal, the results indicate that the secondary structure of BSA molecules was changed in the presence of Intal. PMID- 16707157 TI - Polyhedral oligomeric silsequioxane-polyurethane nanocomposite microvessels for an artificial capillary bed. AB - Fabricating artificial vascularised tissue would involve tissue-engineering techniques, but current technology limits this as cultured cells depend on growth media in vitro and on diffusion in vivo. Therefore, there is a need to construct a synthetic microvascular network, which would sustain these cultured cells in a similar manner to normal tissue. This is again hampered by the poor patency rates of current microvascular grafts. Based on our previous work on polyhedral oligomeric silsesquioxane-polyurethane nanocomposites, which have shown the unique ability to repel coagulant proteins whilst still allowing endothelialisation, we have now developed a new generation of microvascular prosthesis using this polymer. Using these dip-coated nanocomposite microvessels, we have shown that it is possible to mimic the hydraulic conductivity and pressure-responsive radial compliance characteristics of biological microvessels. This would allow nutrient exchange across its walls as well as minimise compliance mismatch throughout the physiological pressure range thus reducing intimal hyperplasia in the long term. This microvessel would have the following implications: (1) as a microvascular substitute to vein grafts and (2) in the future as a component of a microvascular network. PMID- 16707158 TI - Evaluation of mammalian cell adhesion on surface-modified porous silicon. AB - Porous silicon is a promising biomaterial that is non-toxic and biodegradable. Surface modification can offer control over the degradation rate and can also impart properties that promote cell adhesion. In this study, we modified the surface of porous silicon surface by ozone oxidation, silanisation or coating with collagen or serum. For each surface, topography was characterised using atomic force microscopy, wettability by water contact angle measurements, degradation in aqueous buffer by interferometric reflectance spectroscopy and surface chemistry by Fourier-transform infrared spectroscopy. The adhesion of rat pheochromocytoma (PC12) and human lens epithelial cells to these surfaces was investigated. Cells were incubated on the surfaces for 4 and 24 h, and adhesion characteristics were determined by using a fluorescent vital stain and cell counts. Collagen coated and amino silanised porous silicon promoted cell attachment for both cell lines whereas cells attached poorly to ozone oxidised and polyethylene glycol silanised surfaces. We showed that the two cell lines had different adhesion characteristics on the various surfaces at different time points. The use of the vitality assays Alamar Blue (redox based assay) and neutral red (active cellular uptake assay) with porous silicon was also investigated. We reveal incompatibilities between certain resazurin (Alamar Blue), lysosomal incorporation assays (neutral red) and porous silicon. PMID- 16707159 TI - Where's Dad? A need to understand father's role in child maltreatment. PMID- 16707160 TI - Physically abusive fathers and risk assessment. AB - OBJECTIVE: The purpose of this study is to identify factors that predict recidivism among families in which the father is the perpetrator of physical abuse and to compare these factors to the factors that investigators believe are related to higher risk. METHOD: A case-comparison design was used to understand risk among 137 predominantly Caucasian families in which a father had injured a child. RESULTS: The multivariate analysis showed that families in which the father was unemployed (greater time at risk), had younger children, was not the biological father of all of the children, did not take responsibility for his behavior, seriously injured a child, and maltreated a child in the past were more likely to re-abuse. These factors, along with whether the mother had a criminal history, correctly predicted recurrence for 83% of the families. The investigators correctly predicted recurrence for 70% of the families. CONCLUSIONS: The investigators placed too much emphasis on some variables that were not related to recurrence, such as the mother's mental health and whether she tried to protect her children. These results may be useful in adapting assessment systems that do not take into consideration the gender of the perpetrator. PMID- 16707161 TI - The application of in vivo microiontophoresis for the investigation of mast cell neuron interactions in the rat brain. AB - Although mast cells are immune cells of hematopoietic origin, they can be found in parts of the central nervous system of many mammalian species. In the rat brain they are located in the thalamic region. Their function is not defined yet, although they are mostly known to secrete several chemicals, which may influence the surrounding neurons. There are no in vivo electrophysiological data available on the possible effects of brain mast cells on neurons. In this study, we used a combined method of microiontophoresis and extracellular single unit recording to simultaneously activate mast cells and record neuronal action potentials. Four barrelled micropipettes were used for recording neuronal activity and for microiontophoretic application of mast cell degranulator Compound 48/80 (C48/80). Spike sorting routines were performed on-line and off-line to ensure that data were always recorded from a single neuron. C48/80 did not modify the firing rate of cortical neurons (no mast cells are found there), however, it caused excitation (n = 16/37, 43%), or inhibition (n = 9/37, 24%) in thalamic neurons possibly due to mast cell activation. Further investigations will clarify the biochemical nature of changes in neural excitability due to mast cell degranulation in the mammalian brain. PMID- 16707162 TI - Making clinicians lives easier: guidance on use of the QIDS self-report in place of the MADRS. AB - BACKGROUND: The ability to convert total scores from one scale to another facilitates the interpretation of research findings and facilitates the use of systematic measurement in clinical practice. METHODS: Item Response Theory methods were used to convert total scores between the 16-item Quick Inventory of Depressive Symptomatology (QIDS-SR16) and the Montgomery Asberg Depression Rating Scale (MADRS) total scores. Data were obtained from a sample of 233 outpatients with highly treatment-resistant, nonpsychotic major depressive episodes participating in a one-year open label study of vagus nerve stimulation to augment psychotropic medication treatment. RESULTS: MADRS total scores averaged 31.9 (SD = 6.7) at baseline and 21.9 (SD = 11.0) at one year. QIDS-SR16 total scores averaged 17.6 (SD = 3.6) at baseline and 12.5 (SD = 5.8) at one year. Based on one-year data (or exit if the patient did not complete one year), corresponding QIDS-SR16 and MADRS total scores were presented for each possible QIDS-SR16 and MADRS total score. A QIDS-SR16 total score of 5 was comparable to a MADRS total score of 7 or 8 (7.5). LIMITATION: The degree to which these results generalize to less treatment-resistant samples is unknown. CONCLUSION: The conversion of QIDS-SR16 and MADRS total scores provides a basis for clinicians who wish to use the QIDS-SR16 to understand what MADRS total scores reported in clinical trials approximate QIDS-SR16 total scores obtained with their patients. PMID- 16707163 TI - Investigation of genetic association between human Frizzled homolog 3 gene (FZD3) and schizophrenia: results in a Korean population and evidence from meta analysis. AB - The neurodevelopmental hypothesis offers a promising approach through which to explore the pathogenesis of schizophrenia. Human Frizzled-3 protein is one of the major components of the Wnt signaling pathway and plays a crucial role in regulating early neurodevelopmental processes. Therefore, the human Frizzled homolog 3 (FZD3) gene, which encodes this protein, may be an excellent candidate gene for association studies of schizophrenia. To replicate the previously reported positive association between FZD3 and schizophrenia in Han Chinese and Japanese populations, we genotyped two single nucleotide polymorphism (SNP) markers of FZD3 in 241 unrelated schizophrenic patients and 192 control subjects. Fisher's exact test was used to compare the genotypic and allelic frequencies in the two groups. In addition, to integrate and evaluate the accumulated evidence published to date, we performed a meta-analysis on the published data, including our own. No evidence was found to support the association between either of the investigated SNP markers and schizophrenia in a Korean population. Moreover, the meta-analytic result did not support the association between several commonly investigated markers in FZD3 and schizophrenia. Failure to replicate previous positive association results could reflect various theoretical factors. Therefore, other sources of error have to be ruled out before coming to a conclusion. In the case of FZD3, it seems that repeated failures to replicate the original results by several independent research groups combine to provide evidence against an association between FZD3 and schizophrenia. Other candidate genes involved in early neurodevelopmental processes may deserve further investigation. PMID- 16707164 TI - Adrenal cortical responses to low- and high-dose ACTH(1-24) administration in major depressives vs. matched controls. AB - Increased hypothalamo-pituitary-adrenocortical (HPA) axis activity occurs in 30 50% of patients with major depression. This includes normal-to-increased adrenal cortical hormone (cortisol) secretion in spite of reduced corticotropin (ACTH) stimulation. A possible explanation is increased adrenal responsiveness to ACTH. Supporting this possibility is the finding of increased adrenal volume, which reverts to normal with successful treatment. Eight female and six male patients with major depression, and eight female and six male individually matched controls, underwent two test sessions 5-7 days apart. On the first day, a low ACTH(1-24) dose (0.014 microg/kg i.v.), equivalent to 1 microg in a 70-kg individual, was given. On the second day, a supramaximal stimulating dose (250 microg i.v.) was given. Serial blood samples were analyzed for immunoreactive (IR )ACTH, ACTH(1-39), and cortisol. There were no significant sex or patient-control differences in IR-ACTH areas under the curve (AUCs) following low-dose ACTH(1 24), and the correlation between patient and matched control AUCs was +0.71, indicating good correspondence in the amount of circulating ACTH(1-24) available for adrenal stimulation. There were no significant sex or patient-control differences in cortisol response and no significant interaction between dose and subject group, indicating that patients did not differ from controls in their cortisol responses to either low- or high-dose ACTH(1-24). These findings do not indicate increased adrenal cortical responsiveness in patients with major depression. Neurochemical/neurohormonal and neural stimulatory factors other than ACTH might be responsible for the increased adrenal gland size and cortisol secretion, in spite of reduced pituitary ACTH secretion, that has been reported in this illness. PMID- 16707166 TI - Definitive techniques for intractable aspiration pneumonia. PMID- 16707165 TI - A novel antiganglioside specificity against terminal NeuNAc(alfa 2-3)Gal in acute bulbar palsy. AB - We describe a patient with acute oropharyngeal-facial diplegia, tongue palsy and albuminocytological dissociation following upper respiratory tract infection. Electrophysiological abnormalities in blink reflex suggested a brainstem lesion. High titers of anti-GM3, GD1a and GT1b IgG class serum antibodies were initially detected. Absorption studies indicated that antibodies were directed to a common terminal epitope NeuNAc(alfa 2-3)Gal. This novel antiganglioside antibody specificity may play a role in this unusual regional form of acute bulbar palsy of possible central origin. These data are supportive for extending the panel of antiganglioside specificities with anti-GM3. PMID- 16707167 TI - Ear, nose and throat foreign bodies in Melanesian children: an analysis of 1037 cases. AB - OBJECTIVE: Occurrence of foreign bodies (FBs) in ear, nose and throat (ENT) in children are not uncommon in clinical practice. We described our experience with ENT foreign bodies in Melanesian children. METHOD: The study was carried out at the tertiary referral center which is also a University teaching hospital. The 15 year period from 1990 to 2004, 1037 cases of ENT foreign bodies were managed. The clinical, operative and follow up data of these patients were collected from available clinic cards and admission charts. A retrospective analysis was made with these data. RESULTS: Foreign bodies in the external auditory canal constituted 711 (68.6%) cases. It was followed by nose in 258 (24.9%), pharynx in 26 (2.5%), esophagus in 21 (2.0%) and laryngotracheobronchial (LTB) tree in 21 (2.0%) cases. Innumerable varieties of foreign bodies were removed from the ear. The common ones were stones in 277 (39.0%), seeds in 142 (20.0%), plastic ornament beads in 70 (9.8%) and cotton in 48 (6.8%) cases. Six hundred and thirteen (86.2%) of them were removed under microscope at the clinic without any complications. The difficult ones in uncooperative children were removed in the operation theatre. They numbered 98 (13.8%). The nasal foreign bodies were exclusively limited up to the age of 12 years. Common nasal foreign bodies were foam in 54 (20.9%), seeds in 51 (19.8%), stone in 28 (10.9%) and ornament beads in 24 (9.3%) cases. Two hundred and forty-six (95.3%) of them were removed in the clinic. Only 12 (4.7%) were removed in the operation theatre. Fishbone was the commonest (23, 90.2%) foreign body encountered in the pharynx of the children. Coins were the exclusive esophageal foreign bodies in children and all were removed in the operation theatre under general anesthesia. In the LTB tree the FBs were mostly localized to the right main bronchus which occurred in 11 (52.4%) cases. This was followed by the trachea in four (19.1%) cases. The different types of FBs encountered were coffee beans in eight (38.0%), peanuts in five (23.8%), plant seeds in three (14.2%) cases. We encountered two mortalities in these LTB FB patients. CONCLUSIONS: Otolaryngological foreign bodies in children are common. For early diagnosis a high index of suspicion has to be maintained by pediatric otolaryngologist. PMID- 16707168 TI - Two babies, one heart, and no airway. AB - The medical literature on conjoined twins is replete with descriptions of the complexity of surgical separations. However, airway management, although quite challenging, is neglected in literature. We present experience with conjoined twins united anteriorly at the thorax and abdomen sharing one six-chambered heart. Laryngoscopy and intubation recommendations, bronchoscopy techniques, and tracheotomy strategies are outlined for the face-to-face twins. Difficult, unique case report at tertiary care medical center. Conjoined thoraco-omphalopagus twins were successfully intubated at birth using the rigid Seldinger-assisted videotelescopic intubation (SAVI) technique. Flexible airway endoscopy allows for timely management of ventilator-dependent conjoined twins. Tracheotomy was later performed with the twins in the upright position with heads slightly rotated. Meticulous attention to detail, monitoring and vigilance are mandatory. Successful management of conjoined twins relies on close communication and cooperation of all members of the multidisciplinary team. Conjoined thoraco omphalopagus twins united anteriorly require a challenging airway management strategy. A three-dimensional perspective is utilized for intubation and tracheotomy. The SAVI technique is helpful for difficult intubations in the emergent twin airways. PMID- 16707169 TI - Evaluation of 10 commercial diagnostic kits for in vitro expressed hepatitis B virus (HBV) surface antigens encoded by HBV of genotypes A to H. AB - Genetic variability of the hepatitis B virus (HBV) constitutes one of the major challenges for diagnosis of HBV infection. It is plausible that amino acid substitutions in the "a" determinant of the HBV surface antigen (HBsAg) that affect antigenic sites, whether originating from genetic diversity or from mutations in the HBV strain itself, will affect the sensitivity of some diagnostic kits. In fact, recent studies have indicated that some diagnostic kits had false negative results with particular HBsAg mutants. There have been, however, few substantial studies evaluating sensitivities of diagnostic kits to the HBsAg encoded by different HBV genotypes. Our recent study found that 10 diagnostic kits available in Japan were able to detect HBsAg irrespective of whether it originated from HBV genotypes A, B or C, with the latter two genotypes being the dominant species in East Asia. In this study, we extended our previous efforts by assessing the ability of diagnostic kits to detect recombinant HBsAg derived from HBV genotypes A to H. Our results demonstrated that 9 out of 10 diagnostic kits evaluated were able to detect as low as 0.2 International Units (IU)/ml HBsAg, irrespective of HBV genotype. The genotypic differences in the HBV family thus appear to have little impact on the sensitivity of currently available HBsAg diagnostic kits. PMID- 16707170 TI - Preservation of RNA and destruction of infectivity in microdissected brain tissues of Lewis rats infected with the Borna disease virus. AB - Laser microdissection combined with real-time RT-PCR presents an advanced tool to quantify particular RNA species in defined tissue areas. Dealing with infectious tissue samples increases the need to overcome the risk of infectivity and contamination during laser microdissection. Here, an useful method to control infectivity of frozen brain sections infected with the Borna disease virus (BDV), an enveloped RNA virus, is described. Various pre-treatments were applied prior to laser microdissection and subsequent real-time RT-PCR. Brain sections were incubated with Vennotrade mark Vet 1 super 1% or 70% ethanol for 30, 60 and 90min, followed by quantification of infectious virus and RNA recovery using laser microdissection. Total RNA specific for the BDV nucleoprotein (BDV-N) and the cellular genes glyceraldehyde-3-phosphate dehydrogenase (GAPDH), succinate ubiquinone reductase (SDHA) and hypoxanthine phosphoribosyl-transferase-1 (HPRT) was measured by real-time RT-PCR and compared to BDV-infected control samples. After 30 min incubation with both disinfectants, no infectious virus was isolated, while sufficient cDNA copy numbers were amplified. As tissue morphology was best preserved after ethanol treatment, 30min incubation with 70% ethanol was selected as the method of choice to prevent infectivity of BDV. This procedure represents a suitable pre-treatment option to ensure adequate safety of virus infected central nervous system tissue. PMID- 16707172 TI - Behavioral and electrophysiological studies of chronic oral administration of L type calcium channel blocker verapamil on learning and memory in rats. AB - It has been shown that L-type voltage dependent calcium channels (VDCCs) have important role in learning and memory. In vivo and in vitro electrophysiological recordings of hippocampal neurons have demonstrated their involvement in long term potentiation (LTP), which considers being one possible cellular mechanism underlying learning and memory. The long-term effect of VDCCs of hippocampal dentate gyrus (DG) so far on synaptic plasticity has not received much attention. In this study, the effect of chronic (60 days) oral administration of L-type calcium channel blocker verapamil on learning and memory and synaptic plasticity of hippocampal dentate gyrus in rats has been investigated. L-type calcium channel antagonist, verapamil chronically and orally at different doses (10, 20 and 50 mg/kg) was used to investigate learning and memory by passive avoidance learning. LTP in perforant-DG synapses was assessed (by either 200 or 400 Hz tetanization) in order to investigate long-term effect of verapamil on synaptic plasticity. In this case, field excitatory postsynaptic potential (fEPSP) slope and population spike (PS) amplitude were measured. Our behavioral study has shown that chronic oral treatment of verapamil has no effect on learning whereas verapamil (50 mg/kg) decreased memory retrieval. Verapamil (20 and 50 mg/kg) inhibited EPSP-LTP induction at 400 Hz but not at 200 Hz tetanization. Furthermore, only verapamil (50mg/kg) decreased PS-LTP with respect to control group. These data suggest that 400 Hz LTP is required for activation of L-type VDCCs and it seems that verapamil is more effective on L-type calcium channels of DG dendrites than their soma. PMID- 16707171 TI - Behavioral, immunocytochemical and biochemical studies in rats differing in their sensitivity to pain. AB - The aim of the study was to further explore the anatomical and neurochemical background of differences in response to the conditioned aversive stimuli. The different patterns of behavioral coping strategies (a conditioned freezing response and ultrasonic vocalization) were analyzed in animals differing in their response to the acute painful stimulation, a foot-shock (HS: high sensitivity rats, LS: low sensitivity rats, and MS: medium sensitivity rats, according to their behavior in the flinch-jump pre-test), and correlated with plasma corticosterone levels, expression of c-Fos protein, and distribution of 5-HT innervation, in different brain structures. It was found that HS rats showed significantly more freezing behavior, whereas LS animals vocalized much more intensively. The behavior of LS group (less freezing response and stronger vocalization) was related to activation of prefrontal cortex (PFCX), increased activity of adrenal glands and stronger serotonin immunostaining in the PFCX, in comparison with HS animals. The more passive strategy of coping with the aversive event of HS group was related to increased activity of amygdalar nuclei and some areas of the hippocampus, and stronger 5-HT immunostaining in the baso-lateral nucleus of the amygdala, in comparison with LS rats. The present findings suggest that animals more vulnerable to stress might have innate deficits in the activity of brain systems controlling the hypothalamic-pituitary-adrenal axis that would normally allow them to cope with stressful situations. It appears also that response to pain may determine other patterns of emotional behavior, probably reflecting different activation thresholds of some brain structures controlling anxiety, e.g. prefrontal and secondary motor cortex. PMID- 16707173 TI - Quercetin reverses D-galactose induced neurotoxicity in mouse brain. AB - We assessed the neuroprotective effects of quercetin-feeding at doses of 5 and 10 mg/(kg day) on Kunming mice injected daily with D-gal (50 mg/(kg day)) by behavioral tests. Quercetin-fed mice showed higher activity upon induction by new environmental stimuli, lower anxiety and higher novelty-seeking behavior in the open field tasks, and significantly improved learning and memory ability in step through and Morris water Maze tests compared with D-gal-treated mice. We further investigated the mechanisms involved in the neuroprotective effects of quercetin on mouse brain. Quercetin significantly increased superoxide dismutase (SOD) activity and decreased the malondialdehyde (MDA) level. These results imply that quercetin can reverse oxidant impairment induced by D-gal in mouse brain. Neurotoxicity is also associated with Ca(2+) overload induced by oxidant stress. Quercetin could maintain the Ca(2+) homeostasis in the brain of D-gal-treated mice. Furthermore, we also examined the expression of growth-associated protein GAP43 mRNA in mouse brain by in situ hybridization. We found that quercetin dramatically elevated the GAP43 mRNA expression in the brain of D-gal-treated mice to regenerate normal function of neurons against the cellular injury caused by D-gal. PMID- 16707174 TI - Long-term versus intermediate-term supervised exercise training in advanced heart failure: effects on exercise tolerance and mortality. AB - AIM: To date there have been no studies exploring the effects of long-term versus intermediate-term and short-term supervised exercise training program in patients with severe chronic heart failure (CHF) on optimal medical therapy. We assessed exercise tolerance and mortality in CHF patients undergoing long- versus intermediate-term exercise training (ET). METHODS: Forty-two consecutive severe CHF patients (New York Heart Association functional class III) were referred for a supervised exercise and cardiac rehabilitation program and were followed-up for 3 years: 20/42 (48%) patients discontinued ET after intermediate-term period of 1.6+/-0.8 years (Group A, intermediate-term ET), and 22/42 (52%) remained on the ET program for 3.0+/-0.3 years (Group B, long-term ET). Exercise duration, 6-min walked distance and metabolic equivalents (METs) assessed by modified Bruce protocol were recorded before, 4.5 months after, and 3 years after initiation of ET. RESULTS: Both groups were comparable regarding age, gender, prevalence of ischemic etiology, mean ejection fraction and medications. Risk factors for ischemic heart disease were similar, except for the prevalence of diabetes, which was higher in Group A compared to Group B (11/20 versus 5/22, p=0.03). Significantly more Group A patients died after ET discontinuation (4/20 versus 0/22, p=0.01). At the end of follow-up a significant improvement could be seen in Group B patients compared to A in exercise duration, 6-min walked distance and metabolic equivalents (p<0.01 for all). CONCLUSIONS: Higher survival rate was observed in severe CHF patients undergoing long-term versus intermediate-term exercise training. Long-term supervised exercise training is safe and improves exercise tolerance in these patients. PMID- 16707175 TI - Best--worst scaling: What it can do for health care research and how to do it. AB - Statements like "quality of care is more highly valued than waiting time" can neither be supported nor refuted by comparisons of utility parameters from a traditional discrete choice experiment (DCE). Best--worst scaling can overcome this problem because it asks respondents to perform a different choice task. However, whilst the nature of the best--worst task is generally understood, there are a number of issues relating to the design and analysis of a best--worst choice experiment that require further exposition. This paper illustrates how to aggregate and analyse such data and using a quality of life pilot study demonstrates how richer insights can be drawn by the use of best--worst tasks. PMID- 16707176 TI - Accuracy of seed reconstruction in prostate postplanning studied with a CT- and MRI-compatible phantom. AB - BACKGROUND AND PURPOSE: Postimplant dosimetry of prostate seed implants is usually performed by seed localisation on transversal CT or MR images. In order to obtain reliable dosimetric evaluation data, it is important that seeds are reconstructed accurately. Currently, there is no comparative data available on seed localisation accuracy of CT-and MRI-based reconstructions, mainly due to the lack of a suitable QA tool. In this study, we developed a CT-and MRI compatible prostate phantom to investigate the intrinsic accuracy of seed detection for both imaging modalities. PATIENTS AND METHODS: A 60 seed geometry was created according to a clinically meaningful plan, including rotated and shifted seeds. After implantation of the seeds in the phantom, CT and MRI scans with 3, 4 and 5mm slice thickness were performed. The seed locations were reconstructed in the treatment planning system and compared with the known reference positions. RESULTS: Due to the comparable density and relaxation times of the phantom material to prostate tissue, the seeds are visualised similarly as on real patient images. The observed mean reconstruction uncertainties were in general smaller for CT (0.9+/-0.6, 0.9+/-0.6, 2.1+/-0.8 mm on 3, 4 and 5mm scans, respectively), than for MRI (Philips 1.5 T: 2.1+/-1.4, 1.6+/-1.2, 1.9+/-0.9 mm on 3, 4 and 5 mm scans, respectively, and Siemens 1.5 T: 2.3+/-0.8, 2.0+/-1.6, 1.6+/ 0.8 mm on 3, 4 and 5mm scans, respectively). CONCLUSIONS: For our clinical sequences of both CT and MRI, the mean deviation of the reconstructed seed positions were all within acceptable limits for clinical use (<2.3 mm). The phantom was found to be a suitable quality assurance tool to assess the reliability and accuracy of the seed reconstruction procedure. Moreover, as the phantom material has the same imaging characteristics as real prostate tissue, it is a useful device to define proper MRI sequences. PMID- 16707177 TI - The influence of augmented feedback and prior learning on the acquisition of a new bimanual coordination pattern. AB - The present research examined two variables regarding the acquisition of a new bimanual coordination pattern: the role of previous experience and the nature of augmented feedback. Two groups of participants acquired a new coordination pattern (135 degrees relative phase) following two sessions of practice of another novel pattern (90 degrees relative phase). Transfer of learning in these groups was compared to two groups that had not previously learned a new pattern, but were nevertheless influenced by coordination patterns that are intrinsic to the task of bimanual relative timing (in-phase, 0 degrees, and anti-phase, 180 degrees). The findings revealed that new learning overshadowed the influence of the intrinsic patterns. Learning was also greatly affected by augmented feedback: dynamic, on-line pursuit tracking information was more effective in transfer than static, terminal feedback. Implications of these findings regarding theoretical constructs in motor learning are discussed. PMID- 16707178 TI - The effect of viewing the moving limb and target object during the early phase of movement on the online control of grasping. AB - Two experiments were conducted to investigate (1) during which phase of the movement vision is most critical for control, and (2) how vision of the target object and the participant's moving limb affect the control of grasping during that movement phase. In Experiment 1, participants, wearing liquid crystal shutter goggles, reached for and grasped a cylinder with a diameter of 4 or 6 cm under a shutting paradigm (SP) and a re-opening paradigm (RP). In SP, the goggles closed (turned opaque) 0 ms, 150 ms, 350 ms, 500 ms, or 700 ms after movement onset, or remained open (transparent) during the prehension movements. In RP, the goggles closed immediately upon movement onset, and re-opened 0 ms (i.e., without initially shutting), 150 ms, 350 ms, 500 ms, or 700 ms after the initial shutting, or remained opaque throughout the prehension movements. The duration of the prehension movements was kept relatively constant across participants and trials at approximately 1100 ms, i.e., the duration of prehension movements typically observed in daily life. The location of the target object was constant during the entire experiment. The SP and RP paradigms were counter-balanced across participants, and the order of conditions within each session was randomized. The main findings were that peak grip aperture (PGA) in the 150 ms shutting condition was significantly larger than in the 350 ms-shutting condition, and that PGA in the 350 ms-re-opening condition was significantly larger than in the 150 ms-re-opening condition. These results revealed that online vision between 150 ms and 350 ms was critical for grasp control on PGA in typical, daily-life-speeded prehension movements. Furthermore, the results obtained for the time after maximal deceleration (TAMD; movement duration-time to maximal deceleration) demonstrated that early-phase vision contributed to the temporal pattern of the later movement phases (i.e., TAMD). The results thus demonstrated that online vision in the early phase of movement is crucial for the control of grasping. In addition to the apparatus used in Experiment 1, two liquid shutter plates placed in the same horizontal plane (25 cm above the experimental table) were used in Experiment 2 to manipulate the visibility of the target and the participant's moving limb. The plate closest to the participant altered vision of the limb/hand, while the more distant plate controlled vision of the object. The conditions were as follows: (1) both plates were open during movement (full vision condition); (2) both plates were closed 0, 150, or 350 ms following onset of arm movement (front-rear condition: FR); or (3) only the near plate closed 0, 150, or 350 ms following the onset of the arm movement (front condition: F). The results showed that shutting at 0 and 150 ms in the FR condition caused a significantly larger PGA, while the timing of shutting in the F condition had little influence on the PGA. These findings indicated that online vision, especially of the target object, during the early phase of prehension movements is critical to the control of grasping. PMID- 16707179 TI - Motion sickness preceded by unstable displacements of the center of pressure. AB - We exposed standing participants to optic flow in a moving room. Motion sickness was induced by motion that simulated the amplitude and frequency of standing sway. We identified instabilities in displacements of the center of pressure among participants who became sick; these instabilities occurred before the onset of subjective motion sickness symptoms. Postural differences between Sick and Well participants were observed before exposure to the nauseogenic stimulus. During exposure to the nauseogenic stimulus, sway increased for participants who became sick but also for those who did not. However, at every point during exposure sway was greater for participants who became motion sick. The results reveal that motion sickness is preceded by instabilities in displacements of the center of pressure. PMID- 16707180 TI - Survival and growth of Aspergillus carbonarius on wine grapes before harvest. AB - Aspergillus carbonarius, the primary OTA-producing species in Australia, was inoculated onto the surface of Chardonnay and Shiraz bunches at pre-bunch closure, veraison and pre-harvest during the 2002-03 and 2003-04 seasons. Mean A. carbonarius counts decreased between pre-bunch closure and veraison, and increased between veraison and pre-harvest. Increases in A. carbonarius counts from veraison onwards were most marked in Chardonnay bunches during 2003-04; such bunches comprised more berries and were heavier than in 2002-03. Bunches with no berry damage yielded low A. carbonarius counts at pre-harvest and harvest. Exposure to direct sunlight over several days reduced viability of A. carbonarius spores supported on filter membranes by 10(5), despite the spores having thick, heavily melanised walls. The estimated cumulative UV exposure for that period was 10 mWh. Thus, UV radiation may be a contributory factor to the decline of A. carbonarius spores on berry surfaces, particularly in the early stages of berry development. PMID- 16707181 TI - Effect of Lobesia botrana damages on black aspergilli rot and ochratoxin A content in grapes. AB - Sixty-nine wine grape samples of two varieties (Bombino Nero and Uva di Troia) were collected from four vineyards in a high ochratoxin A (OTA) incidence grape growing area in Apulia, southern Italy, during the 2001 and 2002 crop harvests. The levels of toxin, measured in the two year harvests, ranged from 0.02 to 681 ng/g of fresh berries. In both surveys, higher levels of contamination by black aspergilli and OTA were found in both intact and rotten berries originating from bunches damaged by Lobesia botrana larvae as compared to bunches without L. botrana attacks. All berry samples with an OTA level >1 ng/g were contaminated by black aspergilli with a CFU>10(6), and all but one of these samples belonged to the Lobesia damaged berries group. This is the first evidence of an interaction between L. botrana damaged berries and OTA contamination, in field. PMID- 16707182 TI - Ochratoxin A occurrence and formation in Portuguese wine grapes at various stages of maturation. AB - A total of 93 Portuguese grape samples destined for wine production were examined for the presence of ochratoxin A (OTA) and the OTA producing fungi Aspergillus carbonarius and A. niger aggregate. Samples came from 11 vineyards from four winemaking regions in the North and South of the Portuguese mainland, during the harvest seasons of 2001, 2002 and 2003. Grapes were examined at 3 maturation stages, from setting to the harvesting period, to evaluate when contamination with OTA producing fungi and OTA synthesis occur. The detection of fungi in grape samples was made by plating methods with and without surface disinfection. OTA was formed by 14% of the 650 isolates tested. Most of the OTA producing strains (96%) were isolated at harvest time. At this stage, the percentage of grape samples with OTA producing strains detected without surface disinfection was 56%. With surface disinfection, A. carbonarius was isolated from 10% of the samples. OTA was detected in grapes at the 3 maturation stages. The average OTA concentrations in 60 samples at pea berry (28 samples), early veraison (22 samples) and ripe berry (20 samples) were 263, 149 and 35 ng/kg, respectively. Experiments with an A. carbonarius strain demonstrated that OTA production differs significantly with the composition of the berries at different maturation stages (P<0.001), with a mean value of OTA production at pea berry, early veraison and ripe berry of 3402, 1530 and 22 mug/kg, respectively. The production of OTA by A. carbonarius was correlated positively and negatively with the total acidity of grapes (r(s)=0.855, P<0.001) and reducing sugars content (r(s)=-0.835, P<0.001), respectively. Our data demonstrate that OTA synthesis in grapes occurs since early maturation stages. PMID- 16707183 TI - Glycolipids from a colloid chemical point of view. AB - Glycolipids are a group of compounds with a broad range of applications. Two types of glycolipids (alkylpolyglycosides and gangliosides) were examined with regard to their physicochemical properties. Despite their structural differences, they have in common that they are amphiphilic molecules and able to aggregate to form monolayers, bilayers, micelles, lyothropic mesophases or vesicles. The structures of glycolipid micelles were investigated by different experimental techniques in addition to molecular dynamic simulations. The knowledge of the physicochemical properties of gangliosides enables a better understanding of their biological functions. Structural features were obtained for the monosialogangliosides GM1, GM2 and GT1b from bovine brain by means of mass spectrometry. Further the aggregation behaviour was determined by small-angle neutron and dynamic light scattering experiments. Interaction studies of these compounds were carried out by means of surface plasmon resonance using gangliosides incorporated liposomes. They were used as model membranes that interact with the lectins WGA, RCA and HPA. The interaction of lectins immobilized to a modified silicon surface was investigated by in-situ ellipsometry. PMID- 16707184 TI - Expression of alr gene from Corynebacterium glutamicum ATCC 13032 in Escherichia coli and molecular characterization of the recombinant alanine racemase. AB - We constructed the high-expression system of the alr gene from Corynebacterium glutamicum ATCC 13032 in Escherichia coli BL 21 (DE3) to characterize the enzymological and structural properties of the gene product, Alr. The Alr was expressed in the soluble fractions of the cell extract of the E. coli clone and showed alanine racemase activity. The purified Alr was a dimer with a molecular mass of 78 kDa. The Alr required pyridoxal 5'-phosphate (PLP) as a coenzyme and contained 2 mol of PLP per mol of the enzyme. The holoenzyme showed maximum absorption at 420 nm, while the reduced form of the enzyme showed it at 310 nm. The Alr was specific for alanine, and the optimum pH was observed at about nine. The Alr was relatively thermostable, and its half-life time at 60 degrees C was estimated to be 26 min. The K(m) and V(max) values were determined as follows: l alanine to d-alanine, K(m) (l-alanine) 5.01 mM and V(max) 306 U/mg; d-alanine to l-alanine, K(m) (d-alanine) 5.24 mM and V(max) 345 U/mg. The K(eq) value was calculated to be 1.07 and showed good agreement with the theoretical value for the racemization reaction. The high substrate specificity of the Alr from C. glutamicum ATCC 13032 is expected to be a biocatalyst for d-alanine production from the l-counter part. PMID- 16707185 TI - Identification of genistein-inducible and type III-secreted proteins of Bradyrhizobium japonicum. AB - Flagellin is the bulk protein secreted by Bradyrhizobium japonicum. For easier identification of minor protein fractions, the flagellin genes bll6865 and bll6866 were deleted. Extracellular proteins of the corresponding mutant were purified and separated by 2D gel electrophoresis. Several of the protein spots were detectable only after addition of genistein to the growth medium-genistein is an isoflavone secreted by soybean that activates the expression of genes encoding a type III secretion system. These secreted proteins were not present in supernatants of mutants in which conserved genes of the type III secretion system or the regulatory gene ttsI, which is essential for activation of the type III secretion system, are deleted. Out of 22 genistein-inducible protein spots 8 different proteins could be identified by mass spectrometry. One of the proteins, Blr1752, has similarity to NopP of Rhizobium sp. strain NGR234 that is known to be secreted. Another protein is Blr1656 (GunA2) that was shown previously to have endoglucanase activity. Three proteins have similarity to subunits of the flagellar apparatus. Some proteins appeared in several separate spots indicating posttranslational modification. A conserved tts box motif was found in the putative promoter region of six genes encoding secreted proteins. PMID- 16707186 TI - Using TEM to couple transient protein distribution and release for PLGA microparticles for potential use as vaccine delivery vehicles. AB - In the development of tunable PLGA microparticles as vaccine delivery vehicles, it is important to understand the drug distribution within the microparticle over time as well as the long-term release of the drug during polymer degradation. This study addresses the transient 3-D drug distribution in PLGA microparticles during in vitro degradation. Specifically, poly (lactide-co-glycolide) (PLGA 75:25) microparticles containing ovalbumin (OVA) as a model protein were fabricated by double-emulsion (w/o/w) method. The microparticles were incubated at 37 degrees C and 250 rpm in PBS buffer (pH 7.4) over a 100-day period. The in vitro polymer erosion, transient protein distribution profiles and protein release behaviors were investigated. Protein release profiles were determined via spectrophotometry using a BCA assay for the solution. Transmission electron microscopy (TEM) images were obtained for the OVA-loaded microparticles before and during degradation (0 day, 30 days and 60 days), and the corresponding 3-D constructions were developed. From the 3-D constructions, the overall protein distribution of the entire microparticle was vividly reflected. Pixel number analysis of the TEM images was used to quantify transient protein distribution. The transient protein release obtained from the TEM analysis was in good agreement with the BCA analysis. This technique provides an additional tool in helping develop polymer matrices for tunable delivery vehicles in vaccination and other drug delivery scenarios. PMID- 16707187 TI - Anti-angiogenic effects of liposomal prednisolone phosphate on B16 melanoma in mice. AB - Prednisolone phosphate (PLP) encapsulated in long-circulating liposomes can inhibit tumor growth after intravenous administration (i.v.). These antitumor effects of liposomal PLP are the result of the tumor-targeting property of the liposome formulation. The mechanism by which liposomal PLP inhibits tumor growth is unclear. We investigated the effects of liposome-encapsulated PLP versus free PLP on angiogenic protein production in tumor tissue in vivo and on viability and proliferation of tumor and endothelial cells in vitro. In vivo, liposomal PLP had a stronger reducing effect on pro-angiogenic protein levels than free PLP, whereas levels of anti-angiogenic proteins were hardly affected. Cell viability was only slightly affected with either treatment. Liposomal PLP had strong anti proliferative effects on human umbilical vein endothelial cells, whereas free PLP had hardly any effect. Taken together, the present study points to a strong inhibitory effect of liposomal PLP on tumor angiogenesis by reduction of the intratumoral production of the majority of pro-angiogenic factors studied and direct inhibition of endothelial cell proliferation, which is the result of high prolonged levels of prednisolone in the tumor by liposomal delivery. PMID- 16707188 TI - Aminated gelatin microspheres as a nasal delivery system for peptide drugs: evaluation of in vitro release and in vivo insulin absorption in rats. AB - Aminated gelatin microspheres (AGMS) was investigated as a nasal drug delivery system for peptide drugs. The in vitro drug release from microspheres was evaluated using a fluorescein-labeled insulin (RITC-insulin) and FITC-dextran with a molecular weight of 4.4 kDa (FD-4) as model drugs. RITC-insulin release from AGMS was significantly slower than from native gelatin microspheres (GMS), with a cumulate release of 18.4% and 32.4% within 30 min, and 56.9% and 75.1% within 8 h respectively. However, the release of FD-4 from both AGMS and GMS was quite rapid and no difference was observed for the two microspheres. The electrostatic interactions between the model drugs and the microspheres were supposed to be the main factor that controlled the release behavior. The absorption enhancing effect was estimated by measuring the changes of plasma glucose concentrations of healthy rats following intranasal administration of insulin-incorporated microspheres in both suspension and dry powder forms. AGMS could significantly increase the nasal absorption of insulin in rats when administered in a dry powder formulation, but no significant hypoglycemic effect was observed when given in suspensions. One of the mechanisms for the increased insulin absorption was attributed to the hydrogel nature of the microspheres that could absorb water from the nasal mucosa and thus resulted in a temporarily dehydration of the epithelium membrane and opening of the tight junctions. The positive charge of the AGMS has also evidently contributed to the absorption enhancing effect. In addition, the mucoadhesive properties of AGMS might also have played a role to the total effect. AGMS might be a new candidate carrier for the nasal delivery of peptide drugs. PMID- 16707190 TI - SUIT, secretory units of islets in transplantation: An index for therapeutic management of islet transplanted patients and its application to type 2 diabetes. AB - Evaluation of a patient's pancreatic beta-cell function is important in both diagnosis and treatment of diabetes. We sought to determine beta-cell function with a single sampling of blood. Examination of fasting blood glucose (F-BG, mM) and C-peptide (F-CPR, nM) levels in seven post-islet-transplanted states of four patients revealed a linear relationship between F-BG and F-CPR. Assuming that normal subjects aged <40 years have 100% pancreatic beta-cell function, we developed the secretory units of islets in transplantation (SUIT) as an index of beta-cell function by the formula: 250 x F-CPR/(F-BG-3.43). The SUIT index was correlated with the stimulated C-peptide levels not only in islet-transplanted patients (R2 = 0.68, P < 0.05) but also in type 2 patients (R2 = 0.34, P < 0.001). Since the SUIT index can be calculated from data obtained at a single fasting blood sampling and predict the pancreatic beta-cell function, the formula may be a useful tool in clinical management of diabetes. PMID- 16707191 TI - Rapidly increasing diabetes-related mortality with socio-environmental changes in South Korea during the last two decades. AB - Diabetes mellitus is the result of complex interactions involving many genes and environmental factors, and rapid socio-environmental changes have been strongly associated with the development of diabetes. In this study, we examined the trends in diabetes mortality and associated socio-environmental changes that have occurred in South Korea over the last 20 years. Data from a national database and government reports for the years from 1983 to 2001 were analyzed. The data included mortality, socio-economic changes, physical activity and dietary pattern indicators. Deaths from diabetes per 100,000 people increased from 5.3 in 1983 to 18.4 in 2001. Along with increasing diabetes-related mortality, many socio economic indices (gross domestic production, proportion of tertiary industry and urbanization rate), proxies for physical activity (numbers of cars and time spent watching television) and diet indices (animal protein intake and fat intake) showed remarkable changes. To counter increasing prevalence of diabetes and its related mortality in South Korea, multidirectional efforts including lifestyle modification should be mandatory features of future public health policy. PMID- 16707192 TI - Oxytocin alleviates oxidative renal injury in pyelonephritic rats via a neutrophil-dependent mechanism. AB - BACKGROUND: Urinary tract infection (UTI) may cause inflammation of the renal parenchyma and may lead to impairment in renal function and scar formation. Oxidant injury and reactive oxygen species (ROS) have been found responsible in the pathogenesis of UTI. The neurohypophyseal hormone oxytocin (OT) facilitates wound healing and is involved in the modulation of immune and inflammatory processes. We investigated the possible therapeutic effects of OT against Escherichia coli induced pyelonephritis in rats both in the acute and chronic setting. METHODS: Twenty-four Wistar rats were injected 0.1 ml solution containing E. coli ATCC 25922 10(10) colony forming units/ml into left renal medullae. Six rats were designed as sham group and were given 0.1 ml 0.9% NaCl. Pyelonephritic rats were treated with either saline or OT immediately after surgery and at daily intervals. Half of the pyelonephritic rats were decapitated at the 24th hour of E. coli infection, and the rest were followed for 7 days. Renal function tests (urea, creatinine), systemic inflammation markers [lactate dehydrogenase (LDH) and tumor necrosis factor alpha (TNF-alpha)] and renal tissue malondialdehyde (MDA) as an end product of lipid peroxidation, glutathione (GSH) as an antioxidant parameter and myeloperoxidase (MPO) as an indirect index of neutrophil infiltration were studied. RESULTS: Blood urea, creatinine, and TNF alpha levels were increased, renal tissue MDA and MPO levels were elevated and GSH levels were decreased in both of the pyelonephritic (acute and chronic) rats. All of these parameters and elevation of LDH at the late phase were all reversed to normal levels by OT treatment. CONCLUSION: OT alleviates oxidant renal injury in pyelonephritic rats by its anti-oxidant actions and by preventing free radical damaging cascades that involves excessive infiltration of neutrophils. PMID- 16707193 TI - Cocaine- and amphetamine-regulated transcript peptide and sympatho-adrenal axis. AB - Cocaine- and amphetamine-regulated transcript peptide (CART) is constitutively expressed in discrete regions of the mammalian central and peripheral nervous system. Immunohistochemical studies reveal a well-defined network of CART immunoreactive (irCART) neurons organized along the sympatho-adrenal axis. Sympathetic preganglionic neurons, but not parasympathetic preganglionic neurons, in the lateral horn area are CART-positive; which in turn innervate postganglionic neurons in the paravertebral and prevertebral sympathetic ganglia as well as the adrenal medulla. A population of chromaffin cells in the adrenal medulla is CART-positive; whereas, postganglionic neurons are not. Sympathetic preganglionic neurons themselves are contacted by irCART cell processes arising from neurons in the arcuate nucleus, the retrochiasmatic nucleus and the rostral ventrolateral medulla. Results from several recent studies suggest CART directly excites neurons along the sympathetic neural axis or indirectly by potentiating the action of glutamate on NMDA receptors, as evidenced by an elevation of blood pressure and heart rate following intracerebroventricular, intracisternal or intrathecal administration of the peptide to anesthetized rats or conscious rabbits. PMID- 16707194 TI - ESPEN Guidelines on Enteral Nutrition: Liver disease. AB - Enteral nutrition (EN) by means of oral nutritional supplements (ONS) and tube feeding (TF) offers the possibility to increase or to insure nutrient intake in case of insufficient oral food intake. The present guideline is intended to give evidence-based recommendations for the use of ONS and TF in patients with liver disease (LD). It was developed by an interdisciplinary expert group in accordance with officially accepted standards and is based on all relevant publications since 1985. The guideline was discussed and accepted in a consensus conference. EN by means of ONS is recommended for patients with chronic LD in whom undernutrition is very common. ONS improve nutritional status and survival in severely malnourished patients with alcoholic hepatitis. In patients with cirrhosis, TF improves nutritional status and liver function, reduces the rate of complications and prolongs survival. TF commenced early after liver transplantation can reduce complication rate and cost and is preferable to parenteral nutrition. In acute liver failure TF is feasible and used in the majority of patients. PMID- 16707195 TI - Comparison of thiomersal-free and thiomersal-containing formulations of a recombinant hepatitis B vaccine (Hepavax-Gene) in healthy adults. AB - A thiomersal-free (TF) formulation of the recombinant hepatitis B vaccine Hepavax Gene has been developed. This study compared immunogenicity and safety of Hepavax Gene, Hepavax-Gene TF and Engerix-B (containing trace amounts of thiomersal) in a large healthy adult population (N=770) using two vaccination schedules: the priming 0-1-2 months or the standard 0-1-6 month schedule. Hepavax-Gene TF was non-inferior to Hepavax-Gene and Engerix-B with respect to seroprotection rates (>or=10I U/L) 1 month after the third vaccination using the 0-1-6 month schedule, with 99.1% for both Hepavax-Gene formulations and 100% for Engerix-B of subjects' seroprotected in each group. The seroprotection rate after the 0-1-2 schedule was 89.6% for Hepavax-Gene TF, lower than for Hepavax-Gene (94.2%) but comparable to Engerix-B (86.4%). Furthermore, Hepavax-Gene TF was as well tolerated as the comparator vaccines. PMID- 16707196 TI - Immunization with the iron uptake ABC transporter proteins PiaA and PiuA prevents respiratory infection with Streptococcus pneumoniae. AB - Previous studies show that vaccination with the recombinant Streptococcus pneumoniae lipoproteins PiuA and PiaA protects mice against systemic S. pneumoniae disease. The aim of this study was to assess the level of conservation of PiaA and PiuA and a third iron uptake ABC transporter lipoprotein, PitA, between common S. pneumoniae capsular serotypes by sequencing the corresponding genes, and to investigate whether these antigens can protect against respiratory infection. The nucleotide sequences of piuA and piaA were highly conserved in all strains, whereas pitA had significant variation in its nucleotide sequence making PitA an unattractive vaccine candidate. Mucosal vaccination of mice with PiuA and PiaA elicited specific antibody responses in serum and respiratory secretions, and protected against intranasal challenge with S. pneumoniae. These results provide further data indicating that PiuA and PiaA would be suitable candidates for a S. pneumoniae protein antigen vaccine. PMID- 16707197 TI - Dacthal and chlorophenoxy herbicides and chlorothalonil fungicide in eggs of osprey (Pandion haliaetus) from the Duwamish-Lake Washington-Puget Sound area of Washington state, USA. AB - Current-use chlorophenoxy herbicides including 2,4-dichlorophenoxyacetic acid, dicamba, triclopyr, dicamba, dimethyl tetrachloroterephthalate (DCPA or dacthal), and the metabolite of pyrethroids, 3-phenoxybenzoic acid (3-PBA), and the fungicide, chlorothalonil, were investigated in the eggs of osprey (Pandion haliaetus) that were collected from 15 sites from five study areas Puget Sound/Seattle area of Washington State, USA. DCPA differs from acidic chlorophenoxy herbicides, and is not readily hydrolyzed to free acid or acid metabolites, and thus we developed a new method. Of the 12 chlorophenoxy herbicides and chlorothalonil analyzed only DCPA could be quantified at six of these sites (2.0 to 10.3 pg/g fresh weight). However, higher levels (6.9 to 85.5 pg/g fresh weight) of the unexpected DCPA structural isomer, dimethyl tetrachlorophthalate (diMe-TCP) were quantified in eggs from all sites. diMe-TCP concentrations tended to be higher in eggs from the Everett Harbor area. As diMe TCP is not an industrial product, and not commercially available, the source of diMe-TCP is unclear. Regardless, these findings indicate that DCPA and diMe-TCP can be accumulated in the food chain of fish-eating osprey, and transferred in ovo to eggs, and thus may be of concern to the health of the developing chick and the general reproductive health of this osprey population. PMID- 16707198 TI - The effect of winter stress on Ilex aquifolium L.previously fumigated with ozone. AB - European Holly (Ilex aquifolium) received either charcoal-filtered air (CFA) or CFA with 70 nl l(-1) ozone added for 7 h day(-1) over a 28 day period. Plants were then transferred into cooling incubators for hardening (4 degrees C day/2 degrees C night; day length 12 h) for 7 days and then to the frosting stage (2 degrees C day and -5, -10 or -15 degrees C night) for 4 days. The plants were then placed in ambient conditions. Treatment produced significant differences in chlorophyll fluorescence data. Stomatal conductance was significantly higher for the ozone treatments though both showed a general decline over all temperature regimes. Ozone also significantly increased electrolyte leakage and reduced winter survival. These results show that ambient concentrations of ozone can reduce the tolerance of I. aquifolium to freezing stress, which may have serious implications for its establishment and survival. PMID- 16707199 TI - Racial residential segregation and weight status among US adults. AB - While the segmentation of residential areas by race is well known to affect the social and economic well-being of the segregated minority group in the United States, the relationship between segregation and health has received less attention. This study examines the association between racial residential segregation, as measured by the isolation index, and individual weight status in US metropolitan areas. Multi-level, nationally representative data are used to consider the central hypothesis that segregation is positively associated with weight status among African Americans, a group that is hyper-segregated and disproportionately affected by unhealthy weight outcomes. Results show that among non-Hispanic blacks, higher racial isolation is positively associated with both a higher body mass index (BMI) and greater odds of being overweight, adjusting for multiple covariates, including measures of individual socioeconomic status. An increase of one standard deviation in the isolation index is associated with a 0.423 unit increase in BMI (p < 0.01), and a 14% increase in the odds of being overweight (p < 0.01). Among whites, there is no significant association between the isolation index and weight status. These findings suggest that in addition to differences among people, differences among places and, in particular, differences in the spatial organization of persons may be relevant to health policy and promotion efforts. PMID- 16707200 TI - Community participation in AIDS vaccine trials: empowerment or science? AB - For future AIDS vaccines to be successfully tested and implemented, extensive participation in vaccine trials will be necessary. Central to the challenges associated with such participation is concern for protection of participant wellbeing. This short report examines the tension that is sometimes found between the pragmatic need for recruitment into trials, and a more general social good concerning community participation in health. Community empowerment and trial participation are sometimes assumed to go hand in hand, but are potentially contradictory. Recognizing the possible disjunction between empowerment and trial participation allows for clearer discussion of and planning for ethical, scientifically valid trials. PMID- 16707201 TI - Correlation between plasma levels of glutamate, alanine and serine with severity of depression. AB - The goal of this study was to evaluate the utility of using plasma levels of amino acids as an indicator of the severity of depression. The samples were collected from 23 depressed patients receiving antidepressant medication, and were compared to 31 healthy subjects. The plasma levels of amino acids were determined using HPLC with fluorometric detection. The severity of depression was evaluated using the Hamilton Depression Rating Scale (HAM-D) scores. Plasma levels of glutamate, glutamine, glycine and taurine were significantly increased in the depressed patients compared to the controls. Statistical analysis indicated a positive correlation between glutamate and alanine levels and HAM-D scores and a negative correlation of L-serine with HAM-D scores. The results indicate that plasma level of glutamate, alanine and L-serine could reflect the severity of depression rather than glutamine, glycine and taurine. PMID- 16707202 TI - Du-Zhong (Eucommia ulmoides Oliv.) leaves inhibits CCl4-induced hepatic damage in rats. AB - The protective effects of water extract of Du-Zhong (Eucommia ulmoides Oliv.) leaves (WEDZ) and its active compound (protocatechuic acid; PCA) on liver damage were evaluated by carbon tetrachloride (CCl4)-induced chronic hepatotoxicity in rats. Wistar rats were orally treated with WEDZ (0.1, 0.5, and 1.0 g/kg bw) or PCA (0.1 g/kg bw) with administration of CCl4 (0.5 ml/rat, 20% CCl4 in olive oil) for 28 consecutive days. It showed that CCl4-treated rats increased the relative organ weights of liver and kidney. CCl4-induced rats liver damage and significantly (p<0.05) increased the GOT, GPT, LDH and ALP levels in serum as compared with the control group. Treatment with WEDZ or PCA could decrease the GOT, GPT, LDH and ALP levels in serum when compared with CCl4-treated group. CCl4 treated rats also significantly (p<0.05) decreased the GSH content in liver and trolox equivalent antioxidant capacity (TEAC) in serum whereas increased (p<0.05) MDA content in liver as compared with the control group. Treatment with WEDZ or PCA also significantly (p<0.05) increased the GSH content and significantly (p<0.05) decreased the MDA content in liver. Administration of WEDZ or PCA could increase the activities of GPx, GRd and GST in liver. Liver histopathology showed that WEDZ or PCA reduced the incidence of liver lesions including hepatic cells cloudy swelling, lymphocytes infiltration, cytoplasmic vacuolization, hepatic necrosis and fibrous connective tissue proliferated induced by CCl4 in rats. The data suggest that oral administration with WEDZ for 28 consecutive days significantly decrease the intensity of hepatic damage induced by CCl4 in rats. PMID- 16707203 TI - Use of computer-assisted prediction of toxic effects of chemical substances. AB - The current revision of the European policy for the evaluation of chemicals (REACH) has lead to a controversy with regard to the need of additional animal safety testing. To avoid increases in animal testing but also to save time and resources, alternative in silico or in vitro tests for the assessment of toxic effects of chemicals are advocated. The draft of the original document issued in 29th October 2003 by the European Commission foresees the use of alternative methods but does not give further specification on which methods should be used. Computer-assisted prediction models, so-called predictive tools, besides in vitro models, will likely play an essential role in the proposed repertoire of "alternative methods". The current discussion has urged the Advisory Committee of the German Toxicology Society to present its position on the use of predictive tools in toxicology. Acceptable prediction models already exist for those toxicological endpoints which are based on well-understood mechanism, such as mutagenicity and skin sensitization, whereas mechanistically more complex endpoints such as acute, chronic or organ toxicities currently cannot be satisfactorily predicted. A potential strategy to assess such complex toxicities will lie in their dissection into models for the different steps or pathways leading to the final endpoint. Integration of these models should result in a higher predictivity. Despite these limitations, computer-assisted prediction tools already today play a complementary role for the assessment of chemicals for which no data is available or for which toxicological testing is impractical due to the lack of availability of sufficient compounds for testing. Furthermore, predictive tools offer support in the screening and the subsequent prioritization of compound for further toxicological testing, as expected within the scope of the European REACH program. This program will also lead to the collection of high quality data which will broaden the database for further (Q)SAR approaches and will in turn increase the predictivity of predictive tools. PMID- 16707204 TI - Compliance to clinical guidelines for early-stage epithelial ovarian cancer in relation to patient outcome. AB - OBJECTIVES: To assess compliance to current surgical staging and adjuvant treatment guidelines for patients with early-stage epithelial ovarian carcinoma and its impact on overall survival. METHODS: Patients diagnosed between 1991 and 1997 with early-stage ovarian cancer were recruited from the Regional Cancer Registry of the central region in the Netherlands. Demographic data, tumour characteristics, surgical findings and therapeutic data were abstracted from medical records. Patients were classified into optimal and non-optimal surgical staging. Overall survival was estimated using Kaplan-Meier method. To adjust for age hazard ratios for overall survival were estimated with a Cox Proportional Hazards model. RESULTS: One hundred and twenty-five patients were included in the study, 41 of them (32.8%) were optimally staged. Guidelines for adjuvant radio- or chemotherapy were adequately followed in all 62 grade I patients and in 44 out of 59 grade II and III patients (74.6%). During 734.6 person-years of follow up 31 patients died. Five-year overall survival figures were 97.6% in the optimally staged group and 68.5% in the non-optimally staged group. Patients who were non optimally staged, had a significant higher risk to die than those who were optimally staged (HR: 7.4; 95% CI: 1.7-32.2). In patients with a grade II and III tumours, complete surgical staging still had a significant influence on survival (HR: 3.8; 95% CI 1.7-8.3). In women with grade II or III tumours, adjuvant radio- or chemotherapy administered in accordance to the guidelines did not improve overall survival regardless whether they were optimally staged or not. CONCLUSION: Incomplete staging in early-stage ovarian cancer leads to gross mis classification in grade II and III tumours and to a lesser extent in grade I tumours. This leads to undertreatment in both surgical and adjuvant therapy. Subsequently unnecessary deaths may occur. More effort must be put in identifying obstacles interfering with compliance of guidelines. PMID- 16707205 TI - Mechanochemical couplings of kinesin motors. AB - Kinesins are molecular motors capable of moving processively along microtubule in a stepwise manner by hydrolyzing ATP. Numerous experimental results on various aspects of their dynamical behaviours are available in literature. Although a number of models of tightly coordinated mechanism have been proposed to explain some experimental results, up to now no good explanation has been given to all these experimental results by using a single model. We have recently proposed such a model of partially coordinated hand-over-hand moving mechanism. In this paper, we use this model to study in detail various aspects of the dynamical properties of single kinesin molecules. We show that kinesin dimers walk hand over-hand along microtubules in a partially coordinated rather than a tightly coordinated manner. The degree of coordination depends on the ratio of the two heads' ATPase rates that are in turn determined by both internal elastic force and external load. We have tested this model using various available experimental results on different samples and obtained a good agreement between the theory and the experiments. PMID- 16707206 TI - Molecular motion of spin labeled side chains in the C-terminal domain of RGL2 protein: a SDSL-EPR and MD study. AB - Five singly spin labeled side chains at surface sites in the C-terminal domain of RGL2 protein have been analyzed to investigate the general relationship between nitroxide side chain mobility and protein structure. At these sites, the structural perturbation produced by replacement of a native residue with a nitroxide side chain appears to be very slight at the level of the backbone fold. The primary determinants of the nitroxide side chain mobility are backbone dynamics and tertiary interactions. On the exposed surfaces of alpha-helices, the side chain mobility is not restricted by tertiary interactions but appears to be determined by backbone dynamics, while in loop sites, the side chain mobility is even higher. For a better understanding of the changes in the EPR spectral line shape, molecular dynamics simulations were performed and found in agreement with EPR spectral data. PMID- 16707207 TI - Longitudinal dorsal dartos flap for prevention of fistula after a Snodgrass hypospadias procedure. AB - OBJECTIVES: The Snodgrass technique presents the procedure of choice for distal hypospadias. Fistula formation is the most common complication with various rates. We evaluated the importance of a urethral covering using vascularized dorsal subcutaneous tissue for fistula prevention. METHODS: Our study included 126 patients, aged 10 months to 16 years, who underwent hypospadias repair from April 1998 through June 2005. Of the patients, 89 had distal, 30 had midshaft and 7 had penoscrotal hypospadias. All patients underwent standard tubularized incised plate urethroplasty, which was followed by reconstruction of new surrounding urethral tissue. A longitudinal dorsal dartos flap was harvested and transposed to the ventral side by the buttonhole manoeuvre. The flap was sutured to the glans and the corpora cavernosa to completely cover the neourethra with well-vascularized subcutaneous tissue. RESULTS: Mean follow-up was 32 (6-87) months. A successful result without fistula was achieved in all 126 patients. In six patients, temporary stenosis of the glandular urethra occurred and was solved by dilation. CONCLUSIONS: A urethral covering should be performed as part of the Snodgrass procedure. A dorsal well-vascularized dartos flap that is buttonholed ventrally represents a good choice for fistula prevention. Redundancy of the flap and its excellent vascularization depend on the harvesting technique. PMID- 16707208 TI - Re: Richard J. Sylvester, Adrian P.M. van der Meijden, Willem Oosterlinck, J. Alfred Witjes, Christian Bouffioux, Louis Denis and Donald W.W. Newling. Predicting recurrence and progression in individual patients with stage Ta T1 bladder cancer using EORTC risk tables: a combined analysis of 2596 patients from seven EORTC trials. Eur Urol 2006;49:466-77. PMID- 16707209 TI - Estradiol affects androgen-binding protein expression and fertilizing ability of spermatozoa in adult male rats. AB - The estrogenicity of certain environmental pollutants is being increasingly correlated to decline in sperm counts and fertility of the males. Qualitative effects, if any, of estrogen(s) on terminal differentiation of spermatids have been less reported. The present study suggests that exposure to estrogen(s) can also alter the status of condensed chromatin in testicular spermatozoa and reduce their fertilizing potential. A significant reduction was evident in the serum gonadotropins, testosterone, weights of reproductive organs, sperm counts and litters sired by male rats after 10 days of estradiol exposure to a dose of 0.1mg/kg/day. Estradiol treatment led to retardation of in vitro decondensation rates of sperm chromatin, reduction in the uptake of acridine orange dye by chromatin, reduction in susceptibility of chromatin to acid denaturation in vitro, reduced uptake of thiol reactive monobromobimane dye and reduced levels of immunoreactive protamine 1 in caput epididymal sperms. Concomitantly, testicular levels of immunoreactive protamine 1, transition proteins 1/2 and cyclic adenosyl response element modulator-tau (CREMtau) were significantly reduced whilst their mRNA levels were unaffected after estradiol treatment. A significant increase was observed in the testicular mRNA levels of androgen-binding protein (ABP) in estradiol treated sires. An inverse correlation was observed between ABP mRNA levels and uptake of acridine orange by estradiol treated caput sperm chromatin. The results suggest that estradiol-induced increase in ABP mRNA underlies the mechanism(s) involved in the reduction in levels of certain proteins involved in nuclear chromatin condensation during spermiogenesis. PMID- 16707210 TI - Functional neuroanatomy of thyroid hormone feedback in the human hypothalamus and pituitary gland. AB - A major change in thyroid setpoint regulation occurs in various clinical conditions such as critical illness and psychiatric disorders. As a first step towards identifying determinants of these setpoint changes, we have studied the distribution and expression of thyroid hormone receptor (TR) isoforms, type 2 and type 3 deiodinase (D2 and D3), and the thyroid hormone transporter monocarboxylate transporter 8 (MCT8) in the human hypothalamus and anterior pituitary. Although the post-mortem specimens used for these studies originated from patients who had died from many different pathologies, the anatomical distribution of these proteins was similar in all patients. D2 enzyme activity was detectable in the infundibular nucleus/median eminence (IFN/ME) region coinciding with local D2 immunoreactivity in glial cells. Additional D2 immunostaining was present in tanycytes lining the third ventricle. Thyrotropin releasing hormone (TRH) containing neurons in the paraventricular nucleus (PVN) expressed MCT8, TRs as well as D3. These findings suggest that the prohormone thyroxine (T4) is taken up in hypothalamic glial cells that convert T4 into the biologically active triiodothyronine (T3) via the enzyme D2, and that T3 is subsequently transported to TRH producing neurons in the PVN. In these neurons, T3 may either bind to TRs or be metabolized into inactive iodothyronines by D3. By inference, local changes in thyroid hormone metabolism resulting from altered hypothalamic deiodinase or MCT8 expression may underlie the decrease in TRH mRNA reported earlier in the PVN of patients with critical illness and depression. In the anterior pituitary, D2 and MCT8 immunoreactivity occurred exclusively in folliculostellate (FS) cells. Both TR and D3 immunoreactivity was observed in gonadotropes and to a lesser extent in thyrotropes and other hormone producing cell types. Based upon these neuroanatomical findings, we propose a novel model for central thyroid hormone feedback in humans, with a pivotal role for hypothalamic glial cells and pituitary FS cells in processing and activation of T4. Production and action of T3 appear to occur in separate cell types of the human hypothalamus and anterior pituitary. PMID- 16707211 TI - Mutually suppressive interrelations of symmetric epileptic foci in bitemporal epilepsy and their inhibitory stimulation. AB - OBJECTIVES: The goal of this study is to analyze the suppressive interaction of symmetric temporal lobe epileptic foci, assess some failures of epilepsy surgery, and evaluate the possibility of terminating focal seizures with stimulation of symmetric epileptic foci. MATERIALS AND METHODS: One hundred and twenty-nine intractable epilepsy patients (age range 6-53 years) with bitemporal epileptiform abnormalities in multiple scalp EEGs were evaluated with chronically implanted depth and subdural electrodes. Interelectrode coherence and power spectra were studied using internally developed software. RESULTS: Bitemporal epileptic foci were found in 85/129 (66%) patients with reciprocal relations between these foci in 57/85 (67%) patients. Temporal lobectomy was performed for 67/85 patients. 12/67 patients became free of seizures (Engel's Class I), 32/67 improved (Classes II and III), and 23/67 did not improve. 14/23 patients demonstrated post-surgical activation of the contralateral temporal lobe epileptic focus. For 8/14 of these patients, the stereotactic cryoamygdalatomy was performed in the temporal lobe contralateral to the first surgery. 5/8 patients became free of seizures. It was found that stimulation of temporal lobe deep epileptic focus may terminate focal seizures in the contralateral symmetric structures. CONCLUSION: A mutually suppressive relationship is one of variants of the interaction of symmetric epileptic foci. Some epilepsy surgery failures may be a result of post-surgical activation of the intact focus. The increase of coherence between both temporal lobes before the seizure onset of the seizure suggests the establishment of functional interrelations between two epileptic foci at an early, "hidden" phase of seizures, and may predict the direction of seizure spread. Mutually suppressive interrelations of symmetric epileptic foci might be employed for chronic therapeutic stimulation. PMID- 16707212 TI - Photocatalytical degradation of 1,3-dichloro-2-propanol aqueous solutions by using an immobilized TiO2 photoreactor. AB - The photocatalytic oxidation of 1,3-dichloro-2-propanol (1,3-DCP) was studied by following the target compound degradation, the total organic carbon removal rate and by identifying the oxidation products. The reaction was performed in a batch recycle reactor, at room temperature, using UV radiation, H2O2 as oxidant, and immobilized TiO2 as catalyst. 1,3-Dichloro-2-propanone, chloroacetyl-chloride, chloroacetic acid, formic and acetic acid were detected as reaction intermediates and a possible pathway for the oxidation of 1,3-dichloro-2-propanol is proposed. The effect of the oxidative agent's initial concentration was investigated and it was established that higher concentrations of H2O2 slow down the reaction rate. The investigation of the effect of the 1,3-DCP initial concentration showed no influence on the degradation process. The carbon and chloride ion mass balance calculations confirmed the fact that chlorinated intermediates are formed and that they degrade with a lower rate than 1,3-DCP. PMID- 16707213 TI - Oxidation of pesticides by in situ electrogenerated hydrogen peroxide: study for the degradation of 2,4-dichlorophenoxyacetic acid. AB - This paper reports an investigation on the performance of the H2O2 electrogeneration process on a rotating RVC cylinder cathode, and the optimization of the O2 reduction rate relative to cell potential. A study for the simultaneous oxidation of the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) by the in situ electrogenerated H2O2 is also reported. Experiments were performed in 0.3 M of K2SO4, pH of 10 and 3.5. Oxygen concentration in solution was kept in 25 mg L(-1). Maximum hydrogen peroxide generation rate was reached at -1.6 V versus SCE for both, acidic and alkaline solutions. Then, 100 mg L(-1) of 2,4-D was added to the solution. First order apparent rate constants for 2,4-D degradation ranged from 0.9 to 6.3x10(-5) m s(-1), depending on the catalyst used (UV or UV+Fe(II)). TOC reduction was favored in acidic medium where a decreasing of 69% of the initial concentration was observed in the process catalyzed by UV+Fe(II). This figure was an indication that some of the intermediates derived from 2,4-D decomposition remained in solution, mainly as lighter aliphatic compounds. PMID- 16707214 TI - Exploring the interparticle electron transfer process in the photocatalytic oxidation of 4-chlorophenol. AB - This work aimed to investigate the interparticle electron transfer (IPET) process within the coupled-photocatalyst systems on the basis of the degradation of 4 chlorophenol (4-CP). TiO(2), ZnO and SnO(2) are used as the model photocatalysts owing to their increasing energy levels which correspond to the IPET concept. In the single-photocatalyst tests, ZnO tests are associated with the highest degradation rate constants (0.347+/-0.083 h(-1) at pH 7 and 0.453+/-0.011 h(-1) at pH 11) and a better DOC reduction than in other single catalyst tests under given conditions. ZnO/SnO(2) coupled tests have constants of 0.612+/-0.068 and 0.948+/-0.069 h(-1) at pH 7 and 11, respectively. Additionally, the 4-CP prefers the breakdown of chloride group in TiO(2) system while proceeding hydroxylation reaction in ZnO systems. Meanwhile, a phenomenonlogical model coupled with the IPET effect was developed to explore the separation of photo-electrons and photo holes within catalysts. Based on the model parameters, the recombination rate of photo-electrons and photo-holes in TiO(2)/SnO(2) and ZnO/SnO(2) systems is 20-45% lower than that obtained by a respective single catalyst. Thus, coupled photocatalyst tests, TiO(2)/SnO(2) and ZnO/SnO(2) efficiently suppress the recombination, particularly for ZnO/SnO(2) tests at pH 11. PMID- 16707215 TI - Environmental impacts of steel slag reused in road construction: a crystallographic and molecular (XANES) approach. AB - Basic oxygen furnace (BOF) steel slag is a residue from the basic oxygen converter in steel-making operations, and is partially reused as an aggregate for road constructions. Although BOF slag is an attractive building material, its long-term behaviour and the associated environmental impacts must be taken into account. Indeed BOF slag is mainly composed of calcium, silicon and iron but also contains trace amounts of potential toxic elements, specifically chromium and vanadium, which can be released. The present research focuses (i) on the release of Cr and V during leaching and (ii) on their speciation within the bearing phase. Indeed the mobility and toxicity of heavy metals strongly depend on their speciation. Leaching tests show that only low amounts of Cr, present at relatively high concentration in steel slag, are released while the release of V is significantly high. X-ray absorption near-edge structure (XANES) spectroscopy indicates that Cr is present in the less mobile and less toxic trivalent form and that its speciation does not evolve during leaching. On the contrary, V which is predominantly present in the 4+ oxidation state seems to become oxidized to the pentavalent form (the most toxic form) during leaching. PMID- 16707216 TI - The effects of pH on phosphate removal from wastewater by electrocoagulation with iron plate electrodes. AB - In this study, the effect of pH on phosphate removal from wastewater by electrocoagulation with iron plate electrodes has been investigated. For this aim, experiments have been carried out controlled initial pH values within the range of 3-9. Effects of initial pH have been analyzed on efficiencies of phosphate removal and energy consumptions. From obtained results, it was found that optimal initial pH is 3. Besides, experiments have been carried out controlled system pH. Effects of system variables have been analyzed on constant pH. From obtained results in these experiments, it was found that optimal system pH is 7. PMID- 16707217 TI - Incineration of municipal and assimilated wastes in France: assessment of latest energy and material recovery performances. AB - Incineration has an important place in waste management in France. In 2003, around 130 incineration plants have treated 12.6 Mt of non-dangerous waste, mainly composed of household waste (10.8 Mt), non-dangerous waste from industry, business, services (1.0 Mt), sewage sludge (0.2 Mt) or clinical waste (0.1 Mt). The incineration of these wastes generated 3.0 Mt of bottom ash of which 2.3 Mt were used for roads construction and 0.2 Mt of ferrous and non-ferrous metal were recycled. It also produced 2,900,000 MWh of electricity, of which 2,200,000 MWh were sold to Electricite de France (EDF) and 9,100,000 MWh of heat, of which 7,200,000 MWh were sold to private or public users. These French incinerators of non-hazardous waste are currently being thoroughly modernized, thus making possible the consolidation and the enhancement of their environmental and energy performance. This process is related to the implementation of the European Directive 2000/76/CE whose expiration date is 28 December 2005. Upon request of ADEME, the engineering company GIRUS has realised the first technical and economical evaluation of works necessary to bring incinerators into compliance. The financial estimations, carried out in 30 June 2003, show that the investments to be devoted could reach 750 million euros. This assessment shed new light on the situation of non-hazardous waste incinerators, including an identification and a rank ordering for each incinerator of the most frequent and the most complex non-conformities to be solved in term of cost and delay. At last, this assessment gives the solutions for each non-compliance. PMID- 16707218 TI - Apnea patients show a frontopolar inter-hemispheric spindle frequency difference. AB - Sleep apnea syndrome is known to disturb sleep. The purpose of the present work was to study spindle frequency in apnea patients. All-night sleep EEG recordings of 15 apnea patients and 15 control subjects with median ages of 47 and 46 years, respectively, were studied. A previously presented and validated multi-channel spindle analysis method was applied for automatic detection and frequency analysis of bilateral frontopolar and central spindles. Bilateral frontopolar spindles of apnea patients were found to show lower frequencies on the left hemisphere than on the right. Such an inter-hemispheric spindle frequency difference in apnea patients is a novel finding. It could be that the hypoxias and hypercapnias caused by apneic episodes result in local disruption in the regulation of sleep in the frontal lobes. PMID- 16707220 TI - Effects of fatigue of elbow extensor muscles voluntarily induced and induced by electromyostimulation on multi-joint movement organization. AB - To investigate the capacity of the central nervous system to integrate and differentiate two different muscular fatigue states, the present study examines the changes on multi-joint movement organization following muscular fatigue of elbow extensor muscles (triceps brachii) induced by voluntary versus electrically induced contractions. Twenty right-handed male volunteers performed throws in the horizontal plane before and after two fatiguing procedures. First, success rate of throws was not affected by fatigue neither after voluntary contractions, nor after electrically induced contractions. Despite similar reductions of the maximal voluntary isometric force and the median frequency of the electromyographic signal following both fatiguing protocols, voluntary contractions induced greater changes in muscle activation, kinematics and kinetics during throws than electrically induced contractions. The changes observed following voluntary contractions are interpreted as a compensatory strategy involving a greater contribution of the wrist. In contrast, the greater activation of the triceps brachii could compensate the weakness of this muscle induced by fatigue without any modification of the initial multi-joint movement organization. PMID- 16707219 TI - Expression of tumor endothelial marker 7 mRNA and protein in the dorsal root ganglion neurons of the rat. AB - Tumor endothelial marker7 (TEM7) is a transmembrane protein that is highly expressed in the tumor endothelium and neurons. In the present study, the expression profile of TEM7 mRNA and protein was investigated in the dorsal root ganglion (DRG) of the rats. In situ hybridization and reverse transcription polymerase chain reaction studies revealed that TEM7 mRNA expression was localized to the neuronal population of the sensory ganglion. Immunohistochemical analysis on TEM protein with specific antibodies further supported the spatial expression patterns of TEM7 mRNA in the DRG. The temporal expression of TEM7 mRNA in the DRG demonstrated a postnatal increase of TEM7 mRNA expression. Our results indicate that TEM7 may play a role in the peripheral sensory neurons of the vertebrate nervous systems. PMID- 16707221 TI - Variations in amino acid neurotransmitters in the rat ventral spinal cord after hindlimb unloading. AB - We have measured by HPLC the neurotransmitter content in L(4) and L(5) spinal segmental levels in CONT rats, after 7 (HU7) and after 14 days (HU14) of hindlimb unloading. These segments are known to contain the hindlimb muscle motoneurons. The main result is the increase of two neuroexcitators (glutamate and aspartate) and two neuroinhibitors (glycine and GABA) at the L(5) spinal segmental level in HU7 group. Our data indicated that the neurotransmitter changes are restricted to spinal segmental level containing motoneurons from muscles which are strongly modified by HU condition. PMID- 16707222 TI - Evidence-based practice for acute low back pain in primary care: patient outcomes and cost of care. AB - Provider adherence with clinical practice guidelines (CPGs) for acute low back pain (LBP) and its impact on clinical outcomes and cost are controversial. A panel and cross-sectional design was used to determine the rate of provider adherence to the Department of Defense/Veteran's Administration CPG for Acute LBP which included ergonomic and psychosocial intervention. The relationship among adherent care, patient satisfaction, general health, functional outcome, and cost was examined. Direct health services and patient survey data 1 year before and 3 years following guideline implementation were analyzed. The sample consisted of 15,789 LBP cases (CPG) and 14,409 upper extremity cases used as controls (no CPG). Ten to 40% of LBP cases received CPG-adherent care across the 4 years. No differences were found between LBP and upper extremity diagnoses groups in the rates of change in the four outcomes over years. Multivariable regression analyses controlling for demographic variables and perceived health indicated that CPG adherence was related to better functional outcomes, Odds ratio (OR)=1.45 [95% CI=1.31-1.60] and lower healthcare costs (beta=-230.15, p<0.001). Cases receiving more adherent care also reported higher levels of patient satisfaction (beta=0.09, p<0.05) and general health, OR=1.44 [95% CI=1.29-1.60]. Higher levels of adherent care which included efforts to address both ergonomic and psychosocial factors in the course of acute back pain were related to better clinical outcomes and lower costs. PMID- 16707223 TI - The visceromotor responses to colorectal distension and skin pinch are inhibited by simultaneous jejunal distension. AB - Noxious stimuli that are applied to different somatic sites interact; often one stimulus diminishes the sensation elicited from another site. By contrast, inhibitory interactions between visceral stimuli are not well documented. We investigated the interaction between the effects of noxious distension of the colorectum and noxious stimuli applied to the jejunum, in the rat. Colorectal distension elicited a visceromotor reflex, which was quantified using electromyographic (EMG) recordings from the external oblique muscle of the upper abdomen. The same motor units were activated when a strong pinch was applied to the flank skin. Distension of the jejunum did not provoke an EMG response at this site, but when it was applied during colorectal distension it blocked the EMG response. Jejunal distension also inhibited the response to noxious skin pinch. The inhibition of the visceromotor response to colorectal distension was prevented by local application of tetrodotoxin to the jejunum, and was markedly reduced when nicardipine was infused into the local jejunal circulation. Chronic sub-diaphragmatic vagotomy had no effect on the colorectal distension-induced EMG activity or its inhibition by jejunal distension. The nicotinic antagonist hexamethonium suppressed phasic contractile activity in the jejunum, had only a small effect on the inhibition of visceromotor response by jejunal distension. It is concluded that signals that arise from skin pinch and colorectal distension converge in the central nervous system with pathways that are activated by jejunal spinal afferents; the jejunal signals strongly inhibit the abdominal motor activity evoked by noxious stimuli. PMID- 16707224 TI - An experimental study of the contact mode AFM scanning capability of polyimide cantilever probes. AB - This paper presents a preliminary exploration of high-speed contact mode performed with a polyimide probe. The probe is batch micromachined by a lithographic manufacturing process. It offers a spring constant of <0.1N/m, a resonance frequency of about 50k Hz, and a tip diameter of 50-100 nm. The probe is particularly suitable for scanning soft specimens such as biological and polymeric samples. Topographical contact mode imaging at high scanning rates of 48 Hz (1.47 mm/s) has been demonstrated, detecting features <100 nm over a 15 microm scan, yielding >7 bit resolution at 48 Hz. Scanning rates of 16 Hz (0.5mm/s) have been demonstrated for lateral force imaging with spatial resolution of 100 nm over a 15 microm scan, which translates into >7 bit resolution at 16 Hz. These results suggest that the probe can be used in high throughput applications. PMID- 16707225 TI - Debulking surgery for ovarian epithelial cancer performed by a gynaecological oncologist improved survival compared with less specialised surgeons. PMID- 16707226 TI - Effects of D-galactose on the expression of hippocampal peripheral-type benzodiazepine receptor and spatial memory performances in rats. AB - The changes in spatial memory performances and the binding of hippocampal peripheral-type benzodiazepine receptor (PBR) induced by D-galactose (D-gal) were investigated in rats. The animals were randomly divided into two groups: saline treated group and D-gal-induced aging group. All rats received 56 days of injection followed by 5 days of behavioral tests. The D-gal-induced aging rats presented significant impairment in water maze performance, compared with that in the saline-treated rats. A significant decrease in [3H]PK11195 binding in the synaptosomes from hippocampus in the D-gal-induced aging rats was observed, compared to that in the saline-treated rats. Meanwhile, the Scatchard analysis revealed that there was a decrease in Bmax, with no significant change in KD. Further analysis demonstrated that water maze performance was closely related to the PK11195 binding in all rats. These results suggest that D-gal decreased the density of PBR in hippocampal synaptosomes, which may be attributable to the progressive pathogenesis of aging in rats. PMID- 16707227 TI - The cortisol awakening response is blunted in psychotherapy inpatients suffering from depression. AB - Determining the salivary awakening cortisol response (ACR) is a non-invasive, reliable method to detect changes in the hypothalamus-hypopituitary-adrenal (HPA) axis. Although a role of the HPA axis in depression is widely recognized, data on the ACR in depressive patients are still scarce and inconsistent. The present study assessed the ACR in depressed patients admitted for inpatient psychotherapy and a comparison group of other psychiatric diagnoses under the same conditions. The ACR was found to be attenuated in depressed as compared to non-depressed patients. This finding is in contrast to previous studies in healthy subjects or depressed outpatients and suggests a blunted rather than an exacerbated HPA reactivity. Further studies will be needed to disentangle the complex relationship between depression and the ACR. PMID- 16707228 TI - Substance dependency in Iranian patients with hemophilia. AB - AIMS: The prevalence and predisposing factors of substance dependency in 100 Iranian hemophilic patients were investigated. MEASUREMENTS: A confidential questionnaire and DSM-IV criteria were used. Data were gathered from 100 randomly selected hemophilic patients. RESULTS: The mean age of studied patients was 23.42+/-9.67 years, ranging from 12 to 74 years. The rate of substance dependency was high in hemophilic patients (39%), particularly while comparing with normal population. Prevalence of substance dependency was significantly related to age, marital status, number of family members, and being another substance-dependent in the family. Intensity of disease and its associated physical problems were not the main factors moving the patients toward substance dependency. Release of tension and enjoyment were the leading reasons for substance dependency. The nicotine was the most prevalent form of used substance. CONCLUSIONS: The high rate of substance dependency among Iranian hemophilic patients is expected due to the characteristics of disease, loss of psychiatric supports and treatment facilities and the geographic locality of the country. PMID- 16707229 TI - Endothelial nitric oxide-mediated Nrf2 activation as a novel mechanism for vascular and neuroprotection by erythropoietin in experimental subarachnoid hemorrhage. PMID- 16707230 TI - The hyponatramia of multiple myeloma is true and not pseudohyponatramia. AB - The hyponatremia found in multiple myeloma and which is associated with a reduced anion gap (ag) is considered to be pseudohyponatremia due to the displacement of water by the high globulin content in the blood. Serum proteins participate in acid-base balance. Stewart and other authors in their approach to acid-base interpretation acknowledge electrical neutrality as a fundamental characteristic of body fluids. Furthermore, they have shown that both the strong ion difference (SID) and protein, specifically, negatively-charged albumin affect hydrogen ion concentration (H(+)) in the body--i.e., for example an increase in SID leads to a decrease in H(+) and a decrease in albumin leads to a similar effect. The M proteins of multiple myeloma are positively charged. As a result they cause a decrease in sodium levels and the anion gap and thus a true hyponatremia. PMID- 16707231 TI - Improvement of anemia by H. pylori eradication in a patient with myelodysplastic syndrome. PMID- 16707232 TI - Antiinflammatory and ulcerogenic activities of Salvia triloba extracts. AB - In continuation of our research on Salvia triloba, acute and chronic antiinflammatory and ulcerogenic activities of chloroform, ethanol, butanol and water extracts were detected at 25 mg/kg p.o. The chloroform extract showed the highest antiinflammatory activity in both acute and chronic models, while the ulcerogenic effects of all tested extracts were found to be less than that of acetyl salicylic acid. PMID- 16707233 TI - Fungitoxic metabolites from Erigeron apiculatus. AB - Four enyne derivatives (1-4) and quercitrin were isolated during a bioassay guided chromatographic separation of a methanolic extract of Erigeron apiculatus. Matricarialactone (1) and lachnophyllumlactone (2) showed a high fungitoxic activity against Pyricularia oryzae. Matricaria acid methyl ester (3) and lachnophyllum acid methyl ester (4) were, instead, less active. PMID- 16707234 TI - Lethality of essential oil constituents towards the human louse, Pediculus humanus, and its eggs. AB - Essential oils have been widely used in traditional medicine for the eradication of lice, including head lice, but due to the variability of their constitution the effects may not be reproducible. In an attempt to assess the contribution of their component monoterpenoids, a range of common individual compounds were tested in in vitro toxicity model against both human lice (Pediculus humanus, an accepted model of head lice lethality) and their eggs, at different concentrations. No detailed study into the relative potencies of their constituent terpenoids has so far been published. Adult lice were observed for lack of response to stimuli over 3 h and the LT(50) calculated, and the percentage of eggs failing to hatch was used to generate ovicidal activity data. A ranking was compiled for adult lice and partially for eggs, enabling structure activity relationships to be assessed for lethality to both, and showed that, for activity in both life-cycle stages, different structural criteria were required. (+)-Terpinen-4-ol was the most effective compound against adult lice, followed by other mono-oxygenated monocyclic compounds, whereas nerolidol was particularly lethal to eggs, but ineffective against adult lice. PMID- 16707235 TI - [Association of tow thrombotic risk factors: factor V Leiden and hyperhomocysteinemia. A case report]. AB - The identification of constitutional and/or acquired risk factor is of major importance in the treatment of thromboembolic disease in young people; it contributes to evaluate the risk of recurrence and to define the period of oral prophylactic anticoagulant treatment. Several congenital or acquired abnormalities of haemostasis are actually defined. In this paper, we report the case of a 34-year-old man who developed a deep venous thrombosis, five months before the diagnosis of megaloblastic anemia, probably due to pernicious anemia. The thrombosis was partially explained by the acquired hyperhomocysteinemia induced by vitamin B12 deficiency. Moreover, activated protein C resistance due to factor V Leiden, was revealed in our patient. This latter improved under anticoagulant treatment combined with vitamin B12. Combination in one individual, of different risk factors predisposing to inherited and/or acquired thrombophilia, results in increased risk for thrombo-embolic disease, suggesting synergic interaction between these factors. PMID- 16707236 TI - Duration of immunity for canine and feline vaccines: a review. AB - In our studies aimed at assessing the minimum duration of vaccinal immunity (DOI), approximately 1000 dogs have been vaccinated with products from all the major US veterinary biological companies. The DOI for the various products is determined by antibody titers for all dogs and, by challenge studies in selected groups of dogs. Recently, all major companies that make canine vaccines for the U.S. market have completed their own studies; published data show a 3 years or longer minimum DOI for the canine core products, canine distemper virus (CDV), canine parvovirus type 2 (CPV-2), and canine adenovirus-2 (CAV-2). Studies with feline core vaccines - feline parvovirus (FPV), calicivirus (FCV) and herpes virus type I (FHV-1) have shown a minimum DOI of greater than 3 years. Based on these results, the current canine and feline guidelines (which recommend that the last dose of core vaccines be given to puppies and kittens > or =12 weeks of age or older, then revaccination again at 1 year, then not more often than every 3 years) should provide a level of protection equal to that achieved by annual revaccination. In contrast, the non-core canine and feline vaccines, perhaps with the exception of feline leukaemia vaccines, provide immunity for < or =1 year. In general the effectiveness of the non-core products is less than the core products. Thus, when required, non-core vaccines should be administered yearly, or even more frequently. PMID- 16707237 TI - Development of vitamin loaded topical liposomal formulation using factorial design approach: drug deposition and stability. AB - Long-term exposure of the skin to UV light causes degenerative effects, which can be minimized by using antioxidant formulations. The major challenge in this regard is that a significant amount of antioxidant should reach at the site for effective photoprotection. However, barrier properties of the skin limit their use. In the present study, Vitamin E acetate was encapsulated into liposome for improving its topical delivery. However preparation of liposomes is very difficult due to number of formulation variables involved therein. In the present work systematic statistical study for the formulation of liposomes for topical delivery of Vitamin E using the factorial design approach was undertaken. Amount of phospholipid (PL) and cholesterol (CH) were taken at three different levels and liposomes were prepared using ethanol injection method. Liposomes were characterized for encapsulation efficiency, vesicle size, zeta potential, and drug deposition in the rat skin. Gels containing liposomal dispersion (batch with higher skin deposition of VE) were prepared in Carbopol 980 NF and were characterized for gel strength, viscosity and drug deposition in the rat skin. Stability of liposome dispersion and gel formulation was studied at 30 degrees C/65% RH for 3 months. Results of regression analysis revealed that vesicle size and drug deposition in the rat skin were dependent on the lipid concentration and lipid:drug ratio. Drug deposition in rat skin had an inverse relationship with respect to PL and CH concentration. Prepared liposomal dispersion (50 mg PL:6 mg CH) showed seven-fold increase in drug deposition compared to control (plain drug dispersion). Gel formulation demonstrated six-fold and four-fold increase in drug deposition compared to control gel and marketed cream, respectively. Liposome dispersion and gel formulation were found to be stable for 3 months. Factorial design was found to be well suited to identify the key variables affecting drug deposition. Improved drug deposition from liposomal preparations demonstrates its potential for dermal delivery. PMID- 16707238 TI - A study of the use of Ephedra in the manufacture of methamphetamine. AB - The Ephedra plant has been identified as an excellent source of ephedrine and pseudoephedrine, both of which can be chemically reduced to form the widely abused illicit drug methamphetamine. Ephedra contains several additional alkaloids that undergo analogous reductions to form amphetamine and N,N dimethylamphetamine (also drugs of abuse). The main alkaloids obtained from the Ephedra plant have been reduced using four common methods used by the clandestine operator. The intermediates and byproducts of these reductions have been identified and/or tentatively assigned and the mechanism of formation discussed. PMID- 16707239 TI - The detection of sedatives in hair and nail samples using tandem LC-MS-MS. AB - Drug screening methods were developed to detect alprazolam, clobazam, clonazepam, diazepam, midazolam, oxazepam, temazepam, triazolam, zopiclone, and selected metabolites in human hair and nail samples employing liquid-liquid extraction and tandem liquid chromatography-mass spectrometry (LC-MS-MS). Hair and nail samples were obtained from patients who had recently discontinued or were currently prescribed one or more of the targeted drugs. Prazepam was used as the internal standard for all compounds. Some components in the hair matrix gave the same transitions as some of the analytes but did not compromise the analyses because their retention times differed from those for the target compounds. The analytical run time was 8-10min. Results of the hair analysis of a DFSA victim are also presented. PMID- 16707240 TI - Nasal dermoplasty for Japanese hereditary hemorrhagic telangiectasia. AB - OBJECTIVE: While generally considered an effective treatment for moderate to severe epistaxis in hereditary hemorrhagic telangiectasia (HHT), nasal dermoplasty (ND) has not been well established in Japan. This prompted the present Japanese assessment of clinical efficacy and patient satisfaction following this procedure. METHODS: Retrospective analysis of clinical records of 15 patients with HHT undergoing ND between August 1991 and May 2004 and survey of these patients as to postsurgical conditions. Main outcome measures were skin graft "take" frequency after surgery (all patients), reported patient satisfaction (eight recent patients), and reported volume and frequency of epistaxis after versus before surgery (eight recent patients). RESULTS: Graft take rate was 100%. Most patients experienced reduced frequency and volume of bleeding. One patient required an additional operation, total closure of the external nares, 2 years later. Overall patients felt satisfied with ND, experiencing less nasal obstruction than expected. CONCLUSIONS: ND is effective in Japanese patients with moderate and severe nasal bleeding from HHT, reducing their risk of bleeding. PMID- 16707241 TI - Tryptophan enhances ghrelin expression and secretion associated with increased food intake and weight gain in weanling pigs. AB - These studies were conducted to determine whether ghrelin, a 28-amino acid peptide produced mainly by the stomach, was involved in tryptophan-mediated appetite stimulation in swine. In experiment 1, 36 crossbred (Long WhitexLarge White) barrows were used in a 2x3 factorial design to determine the effects of food intake (ad libitum versus limit fed) and tryptophan level (0.12%, 0.19% and 0.26%) on growth performance as well as ghrelin expression, plasma insulin, ghrelin and leptin levels. Ad libitum fed pigs gained more weight, but had poorer feed conversion than limit fed pigs. Weight gain, food intake and feed conversion all improved with increased ingestion of dietary tryptophan. Ad libitum feeding increased plasma insulin. Plasma insulin was unaffected by the level of dietary tryptophan. However, plasma leptin was significantly lower in pigs fed 0.19% tryptophan compared to those fed 0.12% tryptophan. Plasma ghrelin levels and ghrelin mRNA level in gastric fundus and duodenun was significantly higher in pigs fed 0.19% and 0.26% tryptophan diet compared with pigs fed 0.12%. In the second experiment, 18 crossbred barrows were divided into three treatments involving oral infusion of saline, tryptophan (40mg/kg BW) or 5-hydroxytryptophan (40mg/kg BW). Plasma ghrelin levels at 20, 40 and 60min after infusion of tryptophan were higher than after saline and 5-hydroxytryptophan infusion, 5 hydroxytryptophan infusion induced lower food intake than saline infusion, and tryptophan infusion increased food intake 2, 8 and 24h after infusion. In conclusion, oral tryptophan ingestion increased ghrelin expression in gastric fundus and plasma ghrelin level. PMID- 16707242 TI - [Transorbital fiberoptic intubation: a predictable difficult intubation in cephalic surgery]. AB - Management of the difficult adult airway is a crucial problem in anaesthesia. It is the first cause of anaesthetic mortality and morbidity. We report here the case of a patient who could only be intubated through the orbital cavity. We discuss our technique of intubation compared to the other rare procedures described in the literature. We also focus on our anaesthetic protocol and the interest of preserving spontaneous ventilation for intubation. Use of short acting anaesthetic drugs can help to achieve such conditions. PMID- 16707243 TI - Genome-wide analysis, evolutionary expansion, and expression of early auxin responsive SAUR gene family in rice (Oryza sativa). AB - Small auxin-up RNAs (SAURs) are the early auxin-responsive genes represented by a large multigene family in plants. Here, we report the identification of 58 OsSAUR gene family members from rice (Oryza sativa japonica cv Nipponbare), the model monocot plant, by a reiterative database search and manual reannotation; 2 of these are pseudogenes. The coding sequences of OsSAURs do not possess any intron. Most of the predicted OsSAUR protein sequences harbor a putative nuclear localization signal at their N-terminus. Localized gene duplications appear to be the primary genetic event responsible for SAUR gene family expansion in rice. Interestingly, the duplication of OsSAURs was found to be associated with the chromosomal block duplication as well. The phylogenetic analysis revealed that the SAUR gene family expanded in rice and Arabidopsis due to species-specific expansion of the family in monocots and dicots. The auxin-responsive elements and downstream element are conserved in the upstream and downstream sequences, respectively, of OsSAURs. In addition to the 21 OsSAURs with full-length cDNA sequences and 20 with expressed sequence tags, gene expression analyses of at least 7 OsSAURs by RT-qPCR indicated that the majority of identified OsSAURs most likely are expressed in rice. The transcript abundance of the OsSAURs examined increased within a few minutes of exogenous auxin application with varying kinetics. The present study provides basic genomic information for the rice SAUR gene family and will pave the way for deciphering the precise role of SAURs in plant growth and development. PMID- 16707244 TI - Postictal diffusion weighted imaging. AB - PURPOSE: Our aim was to determine whether diffusion weighted imaging can detect abnormalities of diffusivity after single seizures, and investigate the localisation and time course of any changes. METHODS: Twenty-one patients with intractable focal epilepsy were imaged interictally and after 23 seizures. Voxel based statistical parametric mapping was used to detect postictal changes in mean diffusivity (MD), compared to the changes noted in 20 controls scanned twice. The time course and magnitude of the changes were evaluated by measuring MD in the areas of change identified by the voxel-based analysis. RESULTS: Thirty-four focal changes in MD (24 decreases, 10 increases) were detected after 12 of 23 seizures in 11 patients, after a median interval of 53 min from the time of seizure onset. Five patients had areas of both increased and decreased diffusion after seizures. In four patients, postictal changes in diffusion corresponded with the presumed seizure focus. Repeated postictal scanning, after a further interval of a median of 46 min in eight patients, showed that postictal changes in MD, both increases and decreases, were returning towards interictal values. CONCLUSIONS: Diffusion weighted imaging identified focal changes in MD after 52% of single complex partial and secondarily generalised seizures. Changes in MD corresponded to the putative seizure focus in a minority of cases suggesting that the technique is not promising as a method for localising seizure foci, but may indicate the networks involved in seizures. PMID- 16707245 TI - Genetic analysis of the LGI/Epitempin gene family in sporadic and familial lateral temporal lobe epilepsy. AB - Mutations in the LGI1/Epitempin gene cause autosomal dominant lateral temporal lobe epilepsy (ADLTE), a partial epilepsy characterized by the presence of auditory seizures. However, not all the pedigrees with a phenotype consistent with ADLTE show mutations in LGI1/Epitempin, or evidence for linkage to the 10q24 locus. Other authors as well as ourselves have found an internal repeat (EPTP, pfam# PF03736) that allowed the identification of three other genes sharing a sequence and structural similarity with LGI1/Epitempin. In this work, we present the sequencing of these genes in a set of ADLTE families without mutations in both LGI1/Epitempin and sporadic cases. No analyzed polymorphisms modified susceptibility in either the familial or sporadic forms of this partial epilepsy. PMID- 16707246 TI - Selection criteria for antifungals: the right patients and the right reasons. AB - The increasing number of new antifungal agents and their range of licensed indications mean that clinicians now have several pharmacological options for combating fungal infections. This paper briefly reviews the current available agents and seeks to introduce evidence that will help ensure optimal treatment for each type of fungal infection. PMID- 16707247 TI - Effects of oxacillin and tetracycline on autolysis, autolysin processing and atl transcription in Staphylococcus aureus. AB - Autolysins are peptidoglycan hydrolases involved in cell growth and cell lysis. Atl is an important autolysin of Staphylococcus aureus and is essential for penicillin-induced autolysis. The objective of our study was to examine the effect of oxacillin, chloramphenicol and tetracycline on autolysis, peptidoglycan hydrolase profiles and transcription of atl encoding the major S. aureus autolysin on cells grown in the presence of minimum inhibitory concentrations of the antibiotics. Growth of methicillin-susceptible strains in the presence of oxacillin led to increased autolysis, a loss of low molecular weight and a gain of high molecular weight peptidoglycan hydrolase bands suggesting altered proteolytic processing of peptidoglycan hydrolases, and a decrease in atl transcription. In contrast, growth in the presence of tetracycline led to a decrease in autolysis, an increase in atl transcription, and a drastic decrease in the protein concentration of freeze-thaw extracts obtained for peptidoglycan hydrolase analysis. Growth of methicillin-resistant strains in the presence of oxacillin had only moderate effects on autolysis and peptidoglycan hydrolase profiles. PMID- 16707248 TI - The experience is CLEAR. AB - Conventional amphotericin B has been the 'gold standard' for antifungal efficacy, but nephrotoxicity problems limit its clinical utility. In the late 1990s, three lipid-based formulations of amphotericin B were introduced, all of which offered comparable efficacy and reduced renal complications. However, to date, robust safety and comparative efficacy data have been sparse. This paper briefly reviews the available clinical data on amphotericin B lipid complex (ABLC). Also, in detail, it reviews the findings of Collaborative Exchange of Antifungal Research (CLEAR), the most extensive dataset on systemic antifungal treatment with the lipid-based agent ABLC. PMID- 16707249 TI - Chloroquinolines block antibiotic efflux pumps in antibiotic-resistant Enterobacter aerogenes isolates. AB - Efflux mechanisms protect bacterial cells by pumping out toxic compounds and actively contribute to bacterial multidrug resistance. Agents inhibiting efflux pumps are of interest for the control of multidrug-resistant bacterial infections. Herein we report the effects of new chloroquinoline derivatives that render resistant Enterobacter aerogenes isolates noticeably more susceptible to structurally unrelated antibiotics. In addition, some of these chloroquinolines increase the intracellular concentration of chloramphenicol. Some of the molecules tested in this work are able to inhibit the main efflux pump (AcrAB TolC), which is involved in E. aerogenes antibiotic resistance. PMID- 16707250 TI - The changing epidemiology of invasive fungal infections: new threats. AB - The epidemiology of invasive fungal infections is currently at a crucial stage. Potentially fatal fungal strains are emerging that are resistant to many commonly used antifungal agents. It is therefore important to recognize and minimize the major risk factors associated with infection. This paper summarizes the key epidemiological data. PMID- 16707251 TI - What is the current and future status of conventional amphotericin B? AB - Amphotericin B deoxycholate has been the 'gold standard' treatment for invasive fungal infections for over 40 years. Driven to improve on the renal toxicity of amphotericin B deoxycholate, extensive pharmaceutical research has led to the development of several new antifungals including lipid formulations of amphotericin B, broad-spectrum azoles and echinocandins. Compared with amphotericin B deoxycholate, the lipid formulations of amphotericin B (amphotericin B lipid complex, amphotericin B colloidal dispersion and liposomal amphotericin B) share distinct advantages in improved drug safety, in particular reduced incidence and severity of amphotericin B deoxycholate-related nephrotoxicity. However, the lipid formulations of amphotericin B are significantly more expensive than amphotericin B deoxycholate and, as for many of these new antifungals, there are as yet insufficient published studies to guide clinicians. This paper examines aspects of safety, efficacy, and health economic data for the lipid formulations of amphotericin B in particular, in order to provide a rationale to justify substituting amphotericin B deoxycholate with the lipid formulations of amphotericin B. PMID- 16707252 TI - Multidrug resistance to antimicrobials as a predominant factor influencing patient survival. AB - The impact of multidrug resistance to antimicrobials was assessed in a cohort of 243 patients with microbiologically documented infections by a variety of susceptible and multidrug-resistant (MDR) species. Multidrug resistance was defined as resistance to more than two antimicrobial agents of different chemical structure. Cox regression analysis was performed to define differences and the significance of any predisposing factors. Overall survival of patients infected by susceptible isolates was prolonged compared with patients infected by MDR isolates (P=0.013). Mortality rates of infections caused by susceptible and MDR isolates were 4.87% and 16.15%, respectively (P=0.013); the higher mortality rate for MDR isolates was more pronounced for infections by Klebsiella pneumoniae and Pseudomonas aeruginosa. Mean (+/-standard error (S.E.)) survival of patients infected by susceptible and MDR isolates in patients without signs of severe sepsis was 28 days and 27.29+/-0.35 days, respectively (P=not significant). Mean (+/-S.E.) survival of patients with severe sepsis caused by susceptible and MDR isolates was 7.70+/-4.62 days and 10.45+/-2.18 days, respectively (P=0.048). Diabetes mellitus type 2, the presence of severe sepsis and any underlying malignancy were the most important risk factors affecting survival. It is concluded that infections by MDR isolates were accompanied by higher mortality rates and decreased survival compared with infections by susceptible isolates. Diabetes mellitus type 2 and underlying malignancies were significant co-morbid conditions, whereas survival after infection by susceptible isolates was particularly decreased in the event of severe sepsis. PMID- 16707253 TI - The role of brinker in eggshell patterning. AB - Drosophila oogenesis provides a useful system to study signal transduction pathways and their interactions. Through clonal analysis, we found that brinker (brk), a repressor of Dpp signaling, plays an important role in the Drosophila ovary, where its function is essential for dorsal appendage formation. In the absence of brk, operculum fates are specified at the expense of dorsal appendage fates. Brk is expressed by most of the oocyte associated follicle cells, starting from stage 8 of oogenesis. Transforming Growth Factor beta (TGFbeta) signaling represses brk expression in both the early stage egg chambers and in the anterior follicle cells. In brk mutant follicle cell clones at the dorsal anterior region, Broad Complex (BR-C) expression is down-regulated in a larger domain than in wild type. We show that BR-C is required for dorsal appendage development. In large anterior BR-C mutant clones, dorsal appendages are absent, and instead, the eggshell has an enlarged operculum like region at the anterior. In addition, we show that the Epidermal Growth Factor (EGF) receptor signaling represses the TGFbeta signaling in oogenesis by up-regulating brk expression. From our results and previously published data, it appears that anterior follicle cells integrate the levels of EGF receptor activation and TGFbeta receptor activation. Operculum fate results when the sum of the level of activation of both pathways reaches a threshold level, and reduction of activity of one pathway can be compensated to some extent by increase in the other pathway. PMID- 16707254 TI - Suppression of P-glycoprotein expression by antipsychotics trifluoperazine in adriamycin-resistant L1210 mouse leukemia cells. AB - Multidrug resistance (MDR) to unrelated chemotherapeutic drugs can be mediated by overexpression of P-glycoprotein (P-gp), the mdr gene product. Trifluoperazine (TFP), a phenothiazine derivative antipsychotics, is known to reverse MDR of tumor cell lines by blocking P-gp efflux function. In the present study, we evaluated the effect of TFP on the expression of P-gp in multidrug-resistant L1210/Adr mouse leukemic cell lines, which are characterized by overexpession of P-gp. We found that TFP induced the downregulation of P-gp protein and mdr1b mRNA in a dose- and time-dependent manner in L1210/Adr cells. TFP reduction of mdr1b mRNA was paralleled by transcriptional suppression of the mdr1b promoter. Moreover, TFP restored the adriamycin-induced apoptosis in L1210/Adr cells. These results suggest that TFP may have utility as an adjuvant in the therapy of leukemia for the reversal of P-gp-dependent MDR as well as for the management of psychological symptoms in the cancer patients. PMID- 16707255 TI - Antinociceptive activity and chemical composition of constituents from Caragana microphylla seeds. AB - This investigation was undertaken to ascertain the antinociceptive activity of Caragana microphylla Lam. seeds and isolate and characterize the constituents. Antinociceptive activity was screened using acetic acid-induced abdominal constriction in ICR mice. The 75% ethanol extract and some fractions showed analgesic activity, but the antinociceptive activity of chloroform fraction was the strongest and was more productive than other fractions. Seven compounds were isolated from it and identified as: (1) machaeric acid, (2) beta-sitosterol, (3) stigmasterol, (4) pratol, (5) dehydrocavidine, (6) formononetin and (7) sucrose. Caragana microphylla Lam. seeds showed analgesic activity, with the chloroform fraction showing the strongest analgesic activity among the fractions. PMID- 16707256 TI - Protective effect of safranal on pentylenetetrazol-induced seizures in the rat: involvement of GABAergic and opioids systems. AB - The aim of the present study was to evaluate the effects of safranal, an active constituent of Crocus sativus L. stigmas, on seizures induced by pentylenetetrazol. Intracerebroventricular (i.c.v.) microinjection of safranal (4.84, 9.68 and 24.2 micromol) had no effects on tonic and clonic phases as well as mortality upon seizures induced by PTZ (90mg/kg body wt., i.p.). Peripheral administration of safranal (72.75, 145.5 and 291 mg/kg body wt., i.p.), however, induced a dose-dependent decrease in the incidence of both minimal clonic seizures (MCS) (145.5 mg/kg body wt., p<0.01) and generalized tonic-clonic seizures (GTCS) (145.5 mg/kg body wt., p<0.001) following PTZ administration. Safranal also increased MCS and GTCS latency, significantly. Percent of protection against GTCS was 30%, 100% and 100% and mortality protection percent was 40%, 100% and 100% for the mentioned doses, respectively. Pretreatment with flumazenil (5 nmol, i.c.v.) and naloxone (5.5 nmol, i.c.v. and 2 mg/kg body wt., i.p.), 15 min prior to safranal administration (145.5 mg/kg body wt., i.p.), abolished the protective effect of safranal on MCS. Flumazenil also decreased the effect of safranal on incidence as well as latency of GTCS, significantly. These effects were not, however, significant for naloxone (5.5 nmol, i.c.v. and 2mg/kg body wt., i.p.). Results of this study demonstrated that safranal could exert anticonvulsant activity in the PTZ model and this effect may be mediated, at least partly, through GABA(A)-benzodiazepine receptor complex. PMID- 16707257 TI - Influence of the recycled material percentage on the rheological behaviour of HDPE for injection moulding process. AB - The amount of polymer material wasted during thermoplastic injection moulding is very high. It comes from both the feed system of the part, and parts necessary to set up the mould, as well as the scrap generated along the process due to quality problems. The residues are managed through polymer recycling that allows reuse of the materials in the manufacturing injection process. Recycling mills convert the parts into small pieces that are used as feed material for injection, by mixing the recycled feedstock in different percentages with raw material. This mixture of both raw and recycled material modifies material properties according to the percentage of recycled material introduced. Some of the properties affected by this modification are those related to rheologic behaviour, which strongly conditions the future injection moulding process. This paper analyzes the rheologic behaviour of material with different percentages of recycled material by means of a capillary rheometer, and evaluates the influence of the corresponding viscosity curves obtained on the injection moulding process, where small variations of parameters related to rheological behaviour, such as pressure or clamping force, can be critical to the viability and cost of the parts manufactured by injection moulding. PMID- 16707258 TI - mRNPs, polysomes or granules: FMRP in neuronal protein synthesis. AB - mRNA localization and regulated translation play central roles in neurite outgrowth and synaptic plasticity. A key molecule in these processes is the Fragile X mental retardation protein, FMRP, which is involved in the metabolism of neuronal mRNAs. Absence or mutation of FMRP leads to spine dysmorphogenesis and impairs synaptic plasticity. Studies that have mainly been performed on the mouse and Drosophila models for Fragile X Syndrome showed that FMRP is involved in translational regulation at synapses, but even 15 years after discovery of the FMR1 gene, the precise working mechanisms remain elusive. PMID- 16707259 TI - The synaptic vesicle: cycle of exocytosis and endocytosis. AB - Synaptic vesicles are clustered at the presynaptic terminal where they fuse and recycle in response to stimulation. Vesicles appear to be sorted into pools, but we do not yet understand how physiologically defined pools relate to morphological pools. The advent of dynamic imaging approaches has led to an appreciation of the regulation of vesicle mobility. Newly endocytosed vesicles are highly mobile but appear to become transiently trapped as they re-enter the recycling pool. Recent experiments indicate that endocytosis might have a constant rate, but limited capacity. How endocytosis is linked to exocytosis remains unclear, although calcium emerges as an important player. PMID- 16707260 TI - Structures of regulatory elements in mRNAs. AB - Messenger RNAs often contain structures in their 5'-untranslated region that serve to regulate or promote expression of their gene product. Recent structural studies have revealed that riboswitches, which bind a variety of small molecule metabolites, including purine bases, S-adenosylmethionine, amino acids and cofactors, can contain sophisticated tertiary architecture that enables their function, akin to tRNA and rRNA. These structures guide the mRNA to adopt one of two mutually exclusive forms, dictating the outcome of transcription or translation. Another highly structured mRNA element, the viral internal ribosomal entry site, is able to manipulate the ribosome and replace the function of initiation factors to promote gene expression. PMID- 16707261 TI - Autotrophic denitrification and chemical phosphate removal of agro-industrial wastewater by filtration with granular medium. AB - A novel granular medium consisting (1.5-5 mm in diameter) of inert perlite particles as nuclei and an effective surface layer containing sulfur, CaCO3 and Mg(OH)2 was developed for advanced treatment of agro-industrial wastewater. The performance of the medium was examined with a laboratory-scale down-flow fixed bed column reactor using piggery wastewater, which had been treated by an upflow anaerobic sludge blanket reactor and a trickling filter. The removal efficiency of NOx- -N was more than 70% with a NOx- -N loading rate of less than approximately 0.3 kg Nm(-3) d(-1); the removal efficiency dropped due to the accumulation of nitrite when the loading rate exceeded that value. A significant drop of phosphate and Mg2+ concentrations occurred when the effluent pH exceeded 7.9. Ammonium was removed with an average removal efficiency of 12.4%. These results indicated that the crystalline reaction of PO4(3-), Mg2+ and NH4+ (MAP reaction) under alkaline conditions contributed to the removal of phosphate. This medium could be useful for the simultaneous reduction of nitrogenous and phosphorus compounds in biologically treated agro-industrial wastewater. PMID- 16707262 TI - Optimization of nutrients for gellan gum production by Sphingomonas paucimobilis ATCC-31461 in molasses based medium using response surface methodology. AB - A molasses based medium for the production of gellan by Sphingomonas paucimobilis ATCC-31461 was developed. Placket-Burman design criterion was applied to study the effect of various nutrient supplements on gellan production using molasses. Among the 20 variables tested, molasses, tryptone, casaminoacid, disodium hydrogen orthophosphate and manganese chloride showed significant effect on gellan production. A central composite design was applied to determine the optimum concentrations of the significant variables obtained from Placket-Burman design. Most suitable medium composition for production of gellan was (g/l): molasses-112.5; tryptone-1; casaminoacid-1; disodium hydrogen orthophosphate-1; manganese chloride-0.947 and the optimum gellan production was 13.814 g/l. PMID- 16707263 TI - Metacognitive errors in change detection: missing the gap between lab and life. AB - Studies of change detection suggest that people tend to overestimate their ability to detect visual changes. In a recent laboratory study of change detection and human intention, Beck et al., found that individuals have an inadequate understanding that intention can improve change detection performance and that its importance increases with scene complexity. We note that these findings may be specific to unfamiliar situations such as those generated routinely in studies of change detection. In two questionnaire studies, we demonstrate that when participants consider real world scenarios such as driving, people are well aware that the intention to detect changes improves detection performance, especially in complex scenes. We suggest several reasons why change detection findings like Beck et al.'s do not generalize to real world situations. More broadly, we suggest a possible way to bridge the gap between lab and life. PMID- 16707264 TI - Attributes and congruence of three molecular data sets: inferring phylogenies among Septoria-related species from woody perennial plants. AB - To improve our understanding of phylogenetic relationships within the anamorphic genus Septoria, three molecular data sets representing 2,417 bp of nuclear and mitochondrial genes were evaluated. Separate gene analyses and combined analyses were performed using first, the maximum parsimony criterion and second, a Bayesian framework. The homogeneity of data partitions was evaluated via a combination of homogeneity partition tests and tree topology incongruence tests before conducting combined analyses. A last incongruence re-evaluation using partitioned Bremer support was performed on the combined tree, which corroborated the previous estimates. After each separate data set attributes were examined, simple explanations were advocated as the causes of the significant incongruences detected. The analysis of multiple gene partitions showed unprecedented phylogenetic resolution within the genus Septoria that supported the results from previously published single gene phylogenies. Specifically, we have delimited distinct but closely related species representing monophyletic groups that frequently correlated with their respective host families. Conversely, the occurrence of well-supported groups including closely related but distinct molecular taxa sampled on unrelated host-plants allowed us to reject, in these particular cases, the co-evolutionary concept expected between a parasite and its host and to discuss alternative evolutionary models recently proposed for these pathogens. PMID- 16707265 TI - Should we use models to inform policy development? PMID- 16707266 TI - Relaxation and diffusion of perfluorocarbon gas mixtures with oxygen for lung MRI. AB - We report measurements of free diffusivity D(0) and relaxation times T(1) and T(2) for pure C(2)F(6) and C(3)F(8) and their mixtures with oxygen. A simplified relaxation theory is presented and used to fit the data. The results enable spatially localized relaxation time measurements to determine the local gas concentration in lung MR images, so the free diffusivity D(0) is then known. Comparison of the measured diffusion to D(0) will express the extent of diffusion restriction and allow the local surface-to-volume ratio to be found. PMID- 16707267 TI - Molecular dynamics simulations to aid the rational design of organic friction modifiers. AB - Molecular dynamics simulations were performed under conditions of constant volume and temperature and of constant pressure and temperature to elucidate the structure activity relationships of a series of non-ionic surfactant molecules derived from vegetable fat and employed as friction modifiers in commercial engine oils. The simulations show the extent to which intermolecular hydrogen bonding is important in determining the stability of the monolayer formed by the surfactant molecules and show that mono-alkanoyl glyceride molecules are able to pack more efficiently, forming significantly more intermolecular hydrogen bonds and occupying approximately half the volume needed by di-alkanoyl glyceride molecules. Density profiles are presented which show significant mixing of the hydrophobic tail groups and a non-polar solvent. The distribution of torsion angles in the tail groups shows that the conformation is consistent with a liquid at finite temperature rather than a crystal structure. The measured friction coefficients of equimolar solutions of the glycerides show that the efficacy as friction modifiers varies in the order mono-, di- and the tri-oleyl glyceride, which is consistent with the efficacy of film formation predicted by the molecular dynamics calculations. PMID- 16707268 TI - Comparative cytotoxicity of alkyl gallates on mouse tumor cell lines and isolated rat hepatocytes. AB - Alkyl esters of gallic acid inhibited the respiration rate of mouse sarcoma 786A and mouse mammary adenocarcinoma TA3 cell lines and its multiresistant variant TA3-MTX-R more effectively than gallic acid, both in the absence and in the presence of the uncoupler CCCP. The order of inhibition of the respiration rate by gallates in intact cells was n-octyl- approximately iso-amyl- approximately n amyl- approximately iso-butyl->n-butyl->iso-propyl->n-propyl-gallate>>gallic acid. Sarcoma 786A was significantly more susceptible to all seven esters than the TA3 cell line. Respiration rates of the TA3-MTX-R cell line showed almost the same sensitivity to these esters as the TA3 cell line. However, hepatocytes were significantly less sensitive than all tumor cells tested. These alkyl gallates blocked mitochondrial electron flow, mainly at the NADH-CoQ segment, preventing ATP synthesis, which would lead to cellular death. These esters also inhibited, in the same order of potencies as respiration, the growth of 786A, TA3 and TA3 MTX-R cells in culture. In mice carrying TA3 or TA3-MTX-R tumor cells, an important decrease of the tumor growth rate and an increase of survival were observed when mice were treated with iso-butyl gallate alone or in combination with doxorubicin. These results indicate that alkyl gallates are selectively cytotoxic to tumor cells, which may be due to the mitochondrial dysfunctions of these cells. PMID- 16707269 TI - Response of the blood clotting system of the American horseshoe crab, Limulus polyphemus, to a novel form of lipopolysaccharide from a green alga. AB - Lipopolysaccharide (LPS, endotoxin) is a component of Gram-negative bacteria and is the principal indicator to the innate immune systems of higher animals of a Gram-negative bacterial invasion. LPS activates the blood clotting system of the American horseshoe crab, Limulus polyphemus. By stimulating blood cell degranulation, LPS triggers the release of the proteins of the clotting system from the cells, and by activating a protease cascade that converts coagulogen, a soluble zymogen, to coagulin, the structural protein of the clot, LPS triggers the production of the fibrillar coagulin blood clot. Although originally thought to be restricted to the Gram-negative bacteria and the cyanobacteria, LPS, or a very similar molecule, has recently been described from a eukaryotic green alga, Chlorella. Here we show that, like LPS from Gram-negative bacteria, the algal molecule stimulates exocytosis of the Limulus blood cell and the clotting of coagulin. The coagulin clot efficiently entraps the cells of Chlorella in a network of fibrils. Invasion and erosion of the carapace by green algae is an important cause of mortality of Limulus, and it is suggested that the cellular response to aLPS may contribute to defense against this pathogen. PMID- 16707270 TI - Morphometry, ultrastructure, myosin isoforms, and metabolic capacities of the "mini muscles" favoured by selection for high activity in house mice. AB - Prolonged selective breeding of mice (Mus musculus) for high levels of voluntary wheel running has favoured an unusual phenotype ("mini muscles"), apparently caused by a single Mendelian recessive allele, in which most hind-limb muscles are markedly reduced in mass, but have increased mass-specific activities of mitochondrial enzymes. We examined whether these changes reflect changes in fibre size, number or ultrastructure in normal and "mini-muscle" mice within the two (of four) selectively bred lines (lab designations L3 and L6) that exhibit the phenotype at generations 26 and 27. In both lines, the gastrocnemius and plantaris muscles are smaller in mass (by >50% and 20%, respectively) in affected individuals. The mass-specific activities of mitochondrial enzymes in the gastrocnemius and plantaris muscles were increased in the mini phenotype in both lines, with stronger effects in the gastrocnemius muscle. In the gastrocnemius, the % myosin heavy chain (MHC) IIb was reduced by 50% in L3 and by 30% in L6, whereas the % MHC IIa and I were higher, particularly in L3. Fibre number in the plantaris muscle did not significantly differ between mini and normal muscles, although muscle mass was a significant positive correlate of fibre number. Small fibres were more abundant in mini than normal muscles in L3. Mitochondrial volume density was significantly higher in mini than normal muscle fibres in L3, but not in L6. Microscopy revealed a surprising attribute of the mini muscles: an abundance of small, minimally differentiated, myofibril-containing cells positioned in a disorderly fashion, particularly in the surface layer. We hypothesise that these unusual cells may be satellite cells or type IIb fibres that did not complete their differentiation. Together, these observations suggest that mice with the mini phenotype have reduced numbers of type IIb fibres in many of their hind-limb muscles, leading to a decrease in mass and an increase in mass specific aerobic capacity in muscles that typically have a high proportion of type IIb fibres. Moreover, the several statistically significant interactions between muscle phenotype and line indicate that the effect of the underlying allele is altered by genetic background. PMID- 16707271 TI - Thyroid hormone in hypopituitarism, Graves' disease, congenital hypothyroidism, and maternal thyroid disease during pregnancy. AB - Although measurement of serum thyroid-stimulating hormone (TSH) is the single best test to diagnose thyroid disorders and monitor treatment, there are certain situations in which the TSH level cannot be used as a guideline. In the diagnosis of children with hypopituitary hypothyroidism, TSH is often "inappropriately" normal. Treatment is aimed at adjusting the l-thyroxine dose to maintain the serum free thyroxine (T4) in the upper half of the normal range for age. In children with Graves' disease, serum TSH can be suppressed for several months after a euthyroid state is restored, so the clinician must rely on serum free T4 and triiodothyronine (T3) levels. Up to 30% of infants and 10% of children with congenital hypothyroidism have a mildly elevated TSH level despite other indications that the thyroid hormone dosage is correct. Such resistance to thyroid hormone at the pituitary gland diminishes with age. Pregnancy is another condition in which serum TSH levels are altered (slightly lower); in this situation, it is the result of elevated human chorionic gonadotropin levels that cross-react with the TSH receptor. This alteration must be taken into account when diagnosing or treating maternal thyroid disorders during pregnancy. PMID- 16707272 TI - Selective synthesis of sulfoxides from sulfides using ultrasound. AB - The present work aims at developing a new process to selectively synthesize sulfoxide from sulfides using ultrasound. Methyl phenyl sulfide (MPS) has been taken as a model reactant. The reaction has been carried out in ultrasonic bath with an operating frequency of 22 kHz and maximum power supply of 120 W with an actual power dissipation of approximately 40 W measured using calorimetric studies. Effect of various parameters such as presence and absence of catalyst, type of catalyst (catalysts used were sodium tungstate and ammonium molybdate), temperature, concentration of hydrogen peroxide, effect of molar ratio of MPS to H(2)O(2) has been investigated with an aim of obtaining the optimum conditions for the synthesis of sulfoxides. It was observed that the presence of catalyst is a must to achieve appreciable conversions. It was also observed that the periodic addition of stoichiometric amount of hydrogen peroxide instead of sudden addition of it (total quantity of hydrogen peroxide remains the same) gave better selectivity for sulfoxide. The maximum conversion achieved in the present work was approximately 88.42% with sulfone formation being only 0.4% indicating a 99.6% selectivity for the desired product i.e. sulfoxides. PMID- 16707273 TI - Functional role of DMT1 in transferrin-independent iron uptake by human hepatocyte and hepatocellular carcinoma cell, HLF. AB - Non-transferrin bound iron (NTBI) in serum is cleared rapidly by hepatocytes, although the mechanisms of NTBI uptake by hepatocytes are poorly understood. Dietary iron is transported into intestinal enterocytes by divalent metal transporter 1 (DMT1), which also transports iron from transferrin receptor 1 (TfR1)-mediated recycling endosome to intracytoplasm. We made an antiserum against human DMT1 protein derived from mRNA with the iron responsive element (IRE). The DMT1 detected by the antiserum was mainly observed in the membranes of duodenal enterocytes and enterocyte carcinoma (Caco2) cells, whereas DMT1 in normal liver and hepatoma (HLF) cells, was preferentially located in cytoplasm but weakly on cell surface. In addition, iron-depleted HLF increased membrane expression of DMT1, suggesting that the intracellular iron concentration regulated the DMT1 expression in hepatocytes via the iron regulatory protein (IRP)/IRE system. DMT1 overexpressing HLF by DMT1 cDNA transfection expressed DMT1 in both cytoplasm and cell membrane. Although these cells did not change TfR dependent iron uptake, they took up a significant amount of ferrous iron. These results indicate that the DMT1 plays an important role in transporting NTBI into cells. PMID- 16707274 TI - A novel GAL recombinant yeast strain for enhanced protein production. AB - A novel strain of Saccharomyces cerevisiae in which the GAL1 gene was replaced with the GAL4 gene has been designed. The GAL1 gene encodes galactokinase (Gal1p), an enzyme that phosphorylates galactose. Gal4p activates genes necessary for galactose metabolism and is among the best characterized transcription activators. Here we describe a GAL recombinant strain that contains the GAL4 gene fused to the natural GAL1 promoter in addition to the normal constitutively expressed chromosomal GAL4 gene. To evaluate whether both gratuitous induction and regulated overexpression of the positive regulator improve protein production, low- and multi-copy expression vectors containing the GAL1 promoter fused to the structural gene for green fluorescent protein (GFP) were introduced into wild-type, gal1 and GAL recombinant strains. In yeast containing the multi copy plasmid there was an approximately 3.3-fold increase in GFP production in the gal1 mutant strain. Moreover, in the resulting GAL recombinant cells a 4.6 fold increase in fluorescence relative to the wild-type was observed. The GAL recombinant strain should therefore prove useful for maximal expression of heterologous genes driven by a galactose-inducible promoter. PMID- 16707275 TI - Progress towards the development of malaria vaccines. AB - The misery and suffering caused worldwide by infection with the malaria parasite, especially Plasmodium falciparum, has been well documented. Although no licensed vaccine against malaria currently exists, progress has accelerated in recent years towards the goal of developing one. Although the complexity of the malaria parasite has made the malaria vaccine development process tenuous, advances in science and in the vaccine development process as well as increases in funding are encouraging. These advances, coupled with the results of the recent clinical trial of the vaccine candidate RTS,S, have added new vigor to the idea that a malaria vaccine is not only possible but probable. PMID- 16707276 TI - Progress in in silico functional genomics: the malaria Metabolic Pathways database. AB - Genomic, transcriptomic and proteomic data can be turned into biologically meaningful information if they are synthesized into processes. Such amalgamation has been done for the most virulent malaria parasite Plasmodium falciparum in the Metabolic Pathways database. The dialectics of construction of metabolic pathways and other biological processes using this database is presented here. Additional features, such as links to other databases and the incorporation of transcriptomic clocks, are elucidated. Comparison of Metabolic Pathways to other similar databases is analyzed. PMID- 16707277 TI - Evidence based contraceptive choices. AB - People who attend for contraceptive advice have usually formulated an idea of the type of contraceptive that will suit them best. They may wish to use a method that is long, short or medium acting. These are defined as follows: Long-acting method requires renewal no more frequently than every 3 months (e.g. injectable or intrauterine). Short-acting method used daily or with every act of intercourse (e.g. pills, condoms) Medium-acting method requires renewal weekly or monthly (e.g. ring, patch). For men the choice is limited to condoms or vasectomy. Some women do not wish to use hormonal preparations or have an intrauterine device (IUD) or implant inserted. There may also be cultural influences making certain methods of contraception unacceptable. Each of these factors influences the final decision of which method of contraception is decided upon. In addition to taking a full medical and sexual history to identify any risks to the individual's health, which might be increased by a particular contraceptive, time must be spent discussing the options available. It is important to ensure that there is a full understanding of the advantages and disadvantages of each method. The most successful contraceptive method is likely to be the one that the woman (or man) chooses, rather than the one the clinician chooses for them. Access for women to contraception can be improved by having convenient clinic times and service developments such as nurse prescribing and Patient Group Directions. PMID- 16707278 TI - The contribution of spirituality to quality of life in focal epilepsy. AB - People with epilepsy of comparable severity may differ widely in quality of life (QOL), suggesting a role for unexplored individual aspects. This study considered the possible role of spirituality. Thirty-two patients with focal epilepsy completed scales for QOL (World Heath Organization QOL, WHOQOL 100), spirituality (WHO Spirituality, Religiousness, and Personal Beliefs), depression, anxiety, and cognitive efficiency, as well as neuropsychological testing. The QOL and spirituality scales exhibited satisfactory internal consistency. Factor analyses of the scale and test scores yielded separate spiritual (Personal Meaning, Inner Energy, Awe and Transcendence, and Openness), affective (Mood), and cognitive (Cognition, Memory, and Perceived Cognitive Efficiency) factors. The total WHOQOL 100 score was significantly predicted by the Awe and Transcendence and Mood factors. The spiritual, Mood, and Cognition factors significantly predicted single QOL domains. These preliminary results highlight the contribution of spirituality to QOL in epilepsy, encouraging future studies. This could influence the conceptualization and assessment of QOL in these patients. PMID- 16707279 TI - Assemblies of dendrimers and proteins on carbon and gold electrodes. AB - Dendritic macromolecules of two adjacent (G3.5 and G4) generations have been used to modify gold or carbon electrodes. The structure and stability of deposited films have been explored by quartz crystal microbalance (QCM), Surface Plasma Resonance (SPR) and electrochemistry. Dendrimers have been shown to adsorb spontaneously on electrode materials as compressed macromolecular films. They are able to inhibit (G3.5) or promote (G4) electroactive anionic species such as Fe(CN)(6)(3-/4-) used as a probe system. Mixed protein/dendrimer assemblies have been constructed with proteins differing in charge, nature of the prosthetic groups and sizes such as lysozyme, cytochrome c, polyhemic cytochrome c(3) or glucose oxidase. Generally, the stability of adsorbed films seems to be limited to one dendrimer/protein bilayer. Owing to the satisfactory stability of composite cytochrome c(3)/G3.5 or glucose oxidase/G4 films, biosensing applications are described for metal bioremediation and glucose detection, respectively. PMID- 16707281 TI - Detection of eight antibodies in cancer patients' sera against proteins derived from the adenocarcinoma A549 cell line using proteomics-based analysis. AB - To screen cancer for specific autoantibodies, we applied the approach established by Brichory et al., who reported annexins I and II as specific antigens. Solubilized proteins from a cancer cell line (A549) were separated using two dimensional polyacrylamide gel electrophoresis (2D-PAGE), followed by Western blotting (WB) analysis, in which the sera of individual patients were tested for primary antibodies. We found 11 positive spots on PVDF membrane using a WB/enhanced chemiluminescence detection Kit, and identified eight proteins, such as alpha-enolase, inosine-5'-monophosphate dehydrogenase, aldehyde dehydrogenase, 3-phosphoglycerate dehydrogenase, 3-oxoacid CoA transferase, chaperonin, peroxiredoxin 6 and triosephosphate isomerase, that reacted with these antibodies in patients' sera using MALDI-TOF/TOF. All eight antibodies were not detected in the sera derived from lung tuberculosis and healthy controls. PMID- 16707280 TI - Uncoupling protein homologs may provide a link between mitochondria, metabolism and lifespan. AB - Uncoupling proteins (UCPs), which dissipate the mitochondrial proton gradient, have the ability to decouple mitochodrial respiration from ATP production. Since mitochondrial electron transport is a major source of free radical production, it is possible that UCP activity might impact free radical production. Free radicals can react with and damage cellular proteins, DNA and lipids. Accumulated damage from oxidative stress is believed to be a major contributor to cellular decline during aging. If UCP function were to impact mitochondrial free radical production, then one would expect to find a link between UCP activity and aging. This theory has recently been tested in a handful of organisms whose genomes contain UCP1 homologs. Interestingly, these experiments indicate that UCP homologs can affect lifespan, although they do not support a simple relationship between UCP activity and aging. Instead, UCP-like proteins appear to have a variety of effects on lifespan, and on pathways implicated in lifespan regulation. One possible explanation for this complex picture is that UCP homologs may have tissue-specific effects that complicate their effects on aging. Furthermore, the functional analysis of UCP1 homologs is incomplete. Thus, these proteins may perform functions in addition to, or instead of, mitochondrial uncoupling. Although these studies have not revealed a clear picture of UCP effects on aging, they have contributed to the growing knowledge base for these interesting proteins. Future biochemical and genetic investigation of UCP-like proteins will do much to clarify their functions and to identify the regulatory networks in which they are involved. PMID- 16707282 TI - Capsule endoscopy reviewed by a nurse: is it here to stay? PMID- 16707285 TI - Basic life support skills training in a first year medical curriculum: six years' experience with two cognitive-constructivist designs. AB - RATIONALE: Although the Basic Life Support (BLS) ability of a medical student is a crucial competence, poor BLS training programs have been documented worldwide. Better training designs are needed. This study aims to share detailed descriptions and the test results of two cognitive-constructivist training models for the BLS skills in the first year of medical curriculum. METHOD: A BLS skills training module was implemented in the first year curriculum in the course of 6 years (1997-2003). The content was derived from the European Resuscitation Council Guidelines. Initially, a competence-based model was used and was upgraded to a cognitive apprenticeship model in 2000. The main performance-content type that was expected at the end of the course was: competent application of BLS procedures on manikins and peers at an OSCE as well as 60% achievement in a test consisting of 25 MCQ items. A retrospective cohort survey design using exam results and a self-completed anonymous student ratings' questionnaire were used in order to test models. RESULTS: Training time for individual students varied from 21 to 29 hours. One thousand seven hundred and sixty students were trained. Fail rates were very low (1.0-2.2%). The students were highly satisfied with the module during the 6 years. CONCLUSION: In the first year of the medical curriculum, a competence-based or cognitive apprenticeship model using cognitive constructivist designs of skills training with 9 hours theoretical and 12-20 hours long practical sessions took place in groups of 12-17 students; medical students reached a degree of competence to sufficiently perform BLS skills on the manikins and their peers. The cognitive-constructivist designs for skills training are associated with high student satisfaction. However, the lack of controls limits the extrapolation of this conclusion. PMID- 16707283 TI - Effects of stable transfection of human fetal osteoblast cells with estrogen receptor-alpha on regulation of gene expression by tibolone. AB - Tibolone is a synthetic steroid which undergoes tissue selective metabolism into several metabolites having estrogenic, progestogenic or androgenic activities. The effects of 3 alpha-hydroxy tibolone (Org 4094), 3 beta-hydroxy tibolone (Org 30126) and their sulfated metabolites were investigated on human fetal osteoblasts (hFOB). Tibolone had no effect on selected osteoblast marker proteins in estrogen-receptor negative hFOB cells. In contrast, 3 alpha-hydroxy and 3beta hydroxy tibolone resulted in dose-dependent increases in alkaline phosphatase activity in estrogen receptor (ER) alpha-positive hFOB cells. The maximum increase for both metabolites was comparable to the effects of an optimal dose of 17beta-estradiol, and occurred at 10 muM. At 20 muM, both metabolites increased mRNA levels for alkaline phosphatase and type 1 collagen and protein levels for osteocalcin. Sulfated metabolites of tibolone also increased alkaline phosphatase activity. The estrogen receptor antagonist ICI 182, 780 inhibited stimulation of alkaline phosphatase activity by sulfated and non-sulfated tibolone metabolites, but was more potent on the former. Taken together, these results suggest that stable transfection of ER alpha into hFOB cells confers regulation by 3 alpha hydroxy and 3beta-hydroxy tibolone metabolites of osteoblast metabolism. PMID- 16707286 TI - Students' evaluation on a two-stage anatomy curriculum. AB - This study evaluated students' perspectives of the two-stage anatomy course, which is designed to retain the time-honored tradition of cadaver dissection and to include innovative components into anatomy education. A total of 94 sixth-year medical students completed a questionnaire survey at the end of the second stage anatomy course. The results showed that more than half of the students were satisfied with the two-stage anatomy course. Students found that cadaver dissection was most helpful in learning anatomy. The two-stage anatomy course can enhance understanding and memory of anatomical knowledge. However, students disagreed on whether or not the two-stage course can reduce the burden of learning or that the parallel study of anatomy and the 'Life and Death' course could change their attitude toward death more effectively. National Licensing Examination revealed no obvious change after the implementation of the new anatomy course. Traditional and innovative components in anatomy curriculum could coexist in harmony. We provided an alternative for those who wanted to retain cadaver dissection in a reformed curriculum. Further study is needed to evaluate the adequacy of anatomy knowledge and student performance in the long run. PMID- 16707287 TI - Attitudes to concept maps as a teaching/learning activity in undergraduate health professional education: influence of preferred approach to learning. AB - Pre-prepared concept maps that organise knowledge in a non-linear fashion appeal to a variety of cognitive learning styles and may thus represent an educational tool that supports 'teaching to all types'. However, another central cognitive factor, learning approach, may have a bearing on student take-up of this learning resource. Student attitudes to pre-prepared concept maps introduced in Stage 2 MPharm and BSc Pharmacology lectures were therefore examined in relation to the principal learning orientations according to Duff's 30-item revised approaches to study inventory (RASI). Approximately one half of students (49.6 +/- 4.5%) reported pre-prepared concept maps to be useful to their learning (n = 121). When preferred learning approach was examined, derived from the highest RASI score per individual and excluding ties, 31.9 +/- 4.3%, 29.3 +/- 4.2% and 38.8 +/- 4.5% of students demonstrated a preference for the deep approach (DA), strategic approach (STA) and surface approach (SUA), respectively (P > 0.05, chi2 goodness-of-fit test, n = 116). There was a weak but statistically significant association between preferred learning approach identified by Duff's 30-item RASI and the self-reported usefulness of concept maps (P < 0.05, chi2 test of independence; Cramer's V = 0.235; lambda = 0.193). In contrast, gender was not significantly associated with attitude to concept maps in this student cohort. A preliminary analysis of standardised residuals based on observed and expected frequencies revealed that the greatest contributions to this significant association were: a positive influence of DA and a negative influence of STA, respectively, on attitude to concept maps. These data now indicate a contribution of the principal learning orientations vis-a-vis student attitudes to pre-prepared concept maps when employed alongside more traditional teaching/learning activities in medical and biomedical science education, and may further suggest a role for concept maps in the support of deep learning. PMID- 16707288 TI - Introducing peer observation of teaching to GP teachers: a questionnaire study. AB - In medical education programmes which rely on clinical teachers spread across diverse sites, the application of peer observation of teaching offers the potential of both supporting teachers and maintaining quality. This paper reports on a questionnaire survey carried out with general practitioner (GP) teachers of medical undergraduate students from King's College London School of Medicine at Guy's, King's College and St Thomas' Hospitals. The aim of the study was to determine GP teachers' views on a proposed programme of peer observation of their teaching. The majority of GP teachers identified benefits of the proposed scheme with 69% saying it would help improve the education of future doctors. However, despite seeing the benefits, less than half wished to take part in the programme. Two thirds cited time and paperwork as major disincentives to taking part and 62% said that they felt it would make them feel under scrutiny. No associations were found between measures of workload and willingness to take part. This suggests that a fundamental fear of scrutiny and criticism may be the main hurdle to be overcome in implementing the scheme. Imposing peer observation on GP teachers in the form proposed could create suspicion and distance between the university department and practice-based GP teachers and may even result in a loss of teachers. The introduction of peer observation is more likely to be successful if GPs' apprehensions are addressed. Using peer observation to strengthen the process of quality assurance may undermine its role in the support and development of clinical teachers. PMID- 16707289 TI - Reviewing the potential for critical appraisal training to cater for professional practice. AB - Critical appraisal training aims to encourage evidence-based decision-making and ultimately improve health outcomes for patients. Such training must arguably be participatory, multi-disciplinary and problem-based if it is to equip health professionals for problem solving within a modern health service. To explore whether critical appraisal training has the potential to achieve its aims we systematically reviewed reports of critical appraisal training. We identified 58 critical appraisal training programmes, identified through two recently published systematic reviews. Of these, only 15 were identified as multi-disciplinary. Similarly, whilst many of the 58 interventions included some level of participation this was often limited in scope. Around a third of the identified training programmes were problem-based. Only a very small number of the 58 interventions might be described as facilitating cross-disciplinary participatory working. These were by no means all problem-based. We recommend that providers of medical training consider how they might encourage the use of problem-based, mixed, participatory training to encourage evidence-based patient-centered care. More research is also needed to understand how mixed and participatory problem based learning might influence working practice. PMID- 16707290 TI - Dealing with the effect of stress and adverse circumstances on examination results in medical students. PMID- 16707291 TI - BEME systematic review: predictive values of measurements obtained in medical schools and future performance in medical practice. AB - BACKGROUND: Effectiveness of medical education programs is most meaningfully measured as performance of its graduates. OBJECTIVES: To assess the value of measurements obtained in medical schools in predicting future performance in medical practice. METHODS: SEARCH STRATEGY: The English literature from 1955 to 2004 was searched using MEDLINE, Embase, Cochrane's EPOC (Effective Practice and Organization of Care Group), Controlled Trial databases, ERIC, British Education Index, Psych Info, Timelit, Web of Science and hand searching of medical education journals. INCLUSION & EXCLUSIONS: Selected studies included students assessed or followed up to internship, residency and/or practice after postgraduate training. Assessment systems and instruments studied (Predictors) were the National Board Medical Examinations (NBME) I and II, preclinical and clerkship grade-point average, Observed Standardized Clinical Examination scores and Undergraduate Dean's rankings and honors society. Outcome measures were residency supervisor ratings, NBME III, residency in-training examinations, American Specialty Board examination scores, and on-the-job practice performance. DATA EXTRACTION: Data were extracted by using a modification of the BEME data extraction form study objectives, design, sample variables, statistical analysis and results. All included studies are summarized in a tabular form. DATA ANALYSIS AND SYNTHESIS: Quantitative meta-analysis and qualitative approaches were used for data analysis and synthesis including the methodological quality of the studies included. RESULTS: Of 569 studies retrieved with our search strategy, 175 full text studies were reviewed. A total of 38 studies met our inclusion criteria and 19 had sufficient data to be included in a meta-analysis of correlation coefficients. The highest correlation between predictor and outcome was NBME Part II and NBME Part III, r = 0.72, 95% CI 0.30-0.49 and the lowest between NBME I and supervisor rating during residency, r = 0.22, 95% CI 0.13-0.30. The approach to studying the predictive value of assessment tools varied widely between studies and no consistent approach could be identified. Overall, undergraduate grades and rankings were moderately correlated with internship and residency performance. Performance on similar instruments was more closely correlated. Studies assessing practice performance beyond postgraduate training programs were few. CONCLUSIONS: There is a need for a more consistent and systematic approach to studies of the effectiveness of undergraduate assessment systems and tools and their predictive value. Although existing tools do appear to have low to moderate correlation with postgraduate training performance, little is known about their relationship to longer-term practice patterns and outcomes. PMID- 16707292 TI - Systematic review of the literature on assessment, feedback and physicians' clinical performance: BEME Guide No. 7. AB - BACKGROUND AND CONTEXT: There is a basis for the assumption that feedback can be used to enhance physicians' performance. Nevertheless, the findings of empirical studies of the impact of feedback on clinical performance have been equivocal. OBJECTIVES: To summarize evidence related to the impact of assessment and feedback on physicians' clinical performance. SEARCH STRATEGY: The authors searched the literature from 1966 to 2003 using MEDLINE, HealthSTAR, the Science Citation Index and eight other electronic databases. A total of 3702 citations were identified. INCLUSION AND EXCLUSION CRITERIA: Empirical studies were selected involving the baseline measurement of physicians' performance and follow up measurement after they received summaries of their performance. DATA EXTRACTION: Data were extracted on research design, sample, dependent and independent variables using a written protocol. DATA SYNTHESIS: A group of 220 studies involving primary data collection was identified. However, only 41 met all selection criteria and evaluated the independent effect of feedback on physician performance. Of these, 32 (74%) demonstrated a positive impact. Feedback was more likely to be effective when provided by an authoritative source over an extended period of time. Another subset of 132 studies examined the effect of feedback combined with other interventions such as educational programmes, practice guidelines and reminders. Of these, 106 studies (77%) demonstrated a positive impact. Two additional subsets of 29 feedback studies involving resident physicians in training and 18 studies examining proxy measures of physician performance across clinical sites or groups of patients were reviewed. The majority of these two subsets also reported that feedback had positive effects on performance. HEADLINE RESULTS: Feedback can change physicians' clinical performance when provided systematically over multiple years by an authoritative, credible source. CONCLUSIONS: The effects of formal assessment and feedback on physician performance are influenced by the source and duration of feedback. Other factors, such as physicians' active involvement in the process, the amount of information reported, the timing and amount of feedback, and other concurrent interventions, such as education, guidelines, reminder systems and incentives, also appear to be important. However, the independent contributions of these interventions have not been well documented in controlled studies. It is recommended that the designers of future theoretical as well as practical studies of feedback separate the effects of feedback from other concurrent interventions. PMID- 16707293 TI - The impact on students of adverse experiences during medical school. AB - This study aimed to determine the consequences for, and coping method used by, medical students who experienced adverse experiences during their training. A nationwide questionnaire based census of all current medical students in New Zealand. The response rate was 83% (1384/1660). Two-thirds of students had at least one adverse experience, with humiliation being the most common and having the greatest adverse impact. Unwanted sexual advances, unfair treatment on the basis of gender or race had a lesser impact for most students. Most students took several hours or several days to get over an adverse episode and most commonly they then avoided that person or department. Around one half sought help. Only one-quarter felt it motivated their learning while one-sixth felt it made them consider leaving medical school. The most common perpetrators were senior doctors or nurses. Unwanted sexual advances were most common from other students or from patients. Humiliation is the experience that affected students the most and had a significant adverse effect on learning. There is a disturbing rate of unacceptable practice within medical schools, not all of which is from doctors. PMID- 16707294 TI - Medical students' distress--quality, continuity and gender differences during a six-year medical programme. AB - Research observations suggest an increase in distress during the course of medical education, but it is not known whether this distress is chronic and persistent or episodic because follow-ups covering the whole training programme are lacking. We explored stress symptoms among undergraduate medical students (n = 110) at five points during the six-year medical training programme. The quality and continuity of symptoms and gender differences in stress reports were analysed. Questionnaire and interviews were used to assess stress symptoms, perceived health and severity of distress. Stress symptoms, such as fatigue, sleeping problems, anxiety, irritability and depression, were common. No significant gender differences were seen, but there was a consistent increase of stress reports throughout the medical programme in both sexes. Those who were most distressed at the beginning of training also reported more stress later. To conclude, we need interventions that help students to cope with stress, to make a smooth transition from school to medical school, and also to adjust to different learning environments during the different phases of medical education. PMID- 16707295 TI - Design principles for developing an efficient clinical anatomy course. AB - The exponential growth of medical knowledge presents a challenge for the medical school curriculum. Because anatomy is traditionally a long course, it is an attractive target to reduce course hours, yet designing courses that produce students with less understanding of human anatomy is not a viable option. Faced with the challenge of teaching more anatomy with less time, we set out to understand how students employ instructional media to learn anatomy inside and outside of the classroom. We developed a series of pilot programs to explore how students learn anatomy and, in particular, how they combine instructional technology with more traditional classroom and laboratory-based learning. We then integrated what we learned with principles of effective instruction to design a course that makes the most efficient use of students' in-class and out-of-class learning. Overall, we concluded that our new anatomy course needed to focus on transforming how medical students think, reason, and learn. We are currently testing the hypothesis that this novel approach will enhance the ability of students to recall and expand their base of anatomical knowledge throughout their medical school training and beyond. PMID- 16707296 TI - Not just another multi-professional course! Part 2: nuts and bolts of designing a transformed curriculum for multi-professional learning. AB - Multi-professional education has traditionally aimed to develop health professionals who are able to collaborate effectively in comprehensive healthcare delivery. The respective professions learn about their differences in order to work together, rather than developing unity in their commitment to a shared vision of professionalism and service. In this, the second of two papers, the 'nuts and bolts' or practicalities of designing a transformed curriculum for a multi-professional course with a difference is described. Guidelines for the curriculum design process, which seeks to be innovative, grounded in theory and relevant to the learning of the students and the ultimately the health of the patients, include: valuing education; gaining buy-in; securing buy-out; defining of roles; seeking consensus; negotiating difference and expediting decisions. The phases of the design process are described, as well as the educational outcomes envisaged during the process. Reflections of the designers, in particular on what it means to be a multi-professional team, and a reconceptualization of multi professional education are presented as challenges for educators of health professionals. PMID- 16707297 TI - A web-based teaching resource to prepare for final undergraduate examination: a French pilot study. AB - Access to accurate and quality-controlled health information on the Internet for medical students is not an easy task. CISMeF is the search tool of a MeSH-indexed directory of medical Internet resources in French. Since 2004, a new French Pre Residency Examination (PRE) is compulsory for all medical students in the 6th year of the curriculum. The goal of this study is to evaluate CISMeF as a tool to provide teaching resources available on the Internet covering PRE material. The CISMeF terminology and the PRE CISMeF module are described. To assess the CISMeF performance in covering PRE program, its precision (number of relevant resources/number of overall resources extracted by CISMeF) and coverage (number of PRE questions covered by at least one resource in the CISMeF gateway) were computed. The CISMeF module for the new French Pre-Residency Examination is efficient as it already covers 95.7% of the program with a precision of 82.2%. Our data demonstrates that CISMeF is acceptable to guide students' learning and should be a useful teaching resource for the preparation of the French Pre Residency Examination. PMID- 16707298 TI - Using outcomes-based methodology for the education, training and assessment of competence of healthcare professionals. AB - This paper outlines the job competence/functional analysis methodology that is used widely within the UK workforce and throughout the world. This outcomes-based approach to competence focuses on the description of the outcomes of work performance and is not concerned with job or professional titles. The four stages of functional analysis are outlined and illustrated with a functional map developed for the ophthalmic sector. The nature, purpose and format of occupational standards are described and illustrated, along with common errors in developing occupational standards. The knowledge and understanding of requirements that are described with the occupational standards can be used for curriculum development, but the de-linking of standards, learning and assessment means that qualifications can be awarded that are independent of any one learning process, and formal learning programmes are no longer the only access route to the qualification. This approach to competence can support meaningful discussions about how work may be shared between healthcare professions without compromising standards. PMID- 16707299 TI - Continuing Professional Development (CPD) in palliative medicine: a survey. AB - All doctors are required to undertake Continuing Professional development (CPD)- for good practice and for the purposes of annual appraisal and re-validation. The medical workforce in palliative medicine is diverse. Many doctors work on a part time basis, with the same CPD requirements as those working fulltime. This survey was undertaken: to establish whether doctors had fulfilled their CPD requirements in the year 2001-2002 and to identify problems or difficulties experienced by doctors in undertaking CPD. A questionnaire was sent to all doctors known to be working in non-training posts within palliative medicine in October 2002, requesting information on CPD undertaken between 1 April 2001 and 31 March 2002. Questions were also asked regarding knowledge and understanding of appraisal and revalidation. The response rate was 53% (381/721). Palliative medicine was the main post for 276 (72%) respondents and 169 were on the specialist register (44%). The subgroup most likely to have fulfilled CPD requirements were those on the Specialist Register (64%), and those least likely were those holding a contract with a charitable organisation (33%). Less than half of those not on the specialist register understood CPD and revalidation requirements. Overall, only 43% of respondents were meeting CPD requirements (median 25 credits; range 5-375 hours). This study has highlighted widespread barriers to the successful planning, participation in, and recording of, CPD for palliative medicine doctors, with those who are not on the specialist register appearing to be particularly disadvantaged. Individual doctors' information needs need to be addressed, but organisational support for CPD is also required. We believe many of these issues also apply in other specialities. PMID- 16707300 TI - An evaluation of digital, split-site and traditional formats in conference poster sessions. AB - The paper reports an evaluation of digital, split-site and traditional poster presentations at the Association for Medical Education in Europe (AMEE) conference in September 2004. The programme included 300 posters in 19 sessions, viewed, potentially, by 1265 conference participants, in parallel with other events. The instrument was a questionnaire of 16 open- and closed-format questions applied opportunistically and gaining 250 complete responses. Qualitative and quantitative analysis suggested that no one format was preferred. Each had different strengths and weaknesses relating to seeing and hearing the presenter and viewing the poster. Opportunities for discussion were highly valued. PMID- 16707301 TI - Using patient-generated cases to teach students skills in responding to patients' emotions. AB - Responding to patients' emotions has been identified as a core skill in medical interviewing. To give medical students realistic practice in responding to patients' emotions, an exercise was initiated in which simulated patient (SP) cases were developed in collaboration with SPs. Small groups were visited by two SPs, who portray emotional scenarios based on symptoms for which they had previously sought a doctor's care or considered seeking care. SPs also identified circumstances that would provoke emotional reactions for their case. Students and facilitators rated this activity highly. Benefits included more realistic case portrayals, less training time for SPs and more efficient use of SPs. PMID- 16707302 TI - Medical ethics and tomorrow's physicians: an aspect of coverage in the formal curriculum. AB - Medical ethics has created contentious issues and requires reforms in medical education such as renewed emphasis on formal instruction. The aim here was to review the current status of bioethics teaching in medical schools, determine Saudi students' perception of its coverage in the formal curriculum and make recommendations. Using a self-administered questionnaire in a cross-sectional study, undergraduate students' opinion about medical ethics coverage was obtained. Fourteen clinical departments and 201 students were studied. Only 46% of respondents were satisfied with the current coverage of ethical issues in the formal curriculum; 23% were unaware of the value of the subject. Students' approval rate was highest in Neurology and Psychiatry (70%). The study confirmed inadequate formal instruction on medical ethics in a developing country. Five recommendations are made. At admission, students' integrity and character should be assessed. Bioethics should be taught in clinical settings. In the Islamic world, medical curricula should include the Islamic code of medical ethics. Peers, nurses and patients should evaluate graduates' performance in ethics at the bedside. Evidence-based assessment and continuous quality improvement are required to maintain the requisite standard. PMID- 16707303 TI - A sexual history-taking curriculum for second year medical students. AB - The purpose of this study is to describe the evaluation of a sexual history taking curriculum and correlates of student performance during a Clinical Skills Assessment. Reading assignments, small group discussions, a Saturday Sex workshop and performance on a Clinical Skills Assessment were evaluated. Students most favorably rated the workshop and least favorably rated the reading assignments. Eighty-four percent of students asked at least one sexual history question on the Clinical Skills Assessment. We were unable to identify any independent predictors of sexual history-taking behavior. PMID- 16707304 TI - Internationalization of medical education in the Netherlands: state of affairs. AB - In the framework of the Bologna Process, internationalization co-ordinators of seven (out of eight) Dutch medical schools completed an electronic survey about internationalization-related aspects of the curriculum. Common features of internationalization in Dutch medical schools were: the numbers of outgoing students exceeded the numbers of incoming students, and most international programmes involved clinical training and research projects. We recommend that Dutch medical schools should pay more attention to 'Internationalization at Home' and focus on conditions that are conducive to participation by foreign students. PMID- 16707305 TI - First aid and basic life support of junior doctors: A prospective study in Nijmegen, the Netherlands. AB - According to the Dutch medical education guidelines junior doctors are expected to be able to perform first aid and basic life support. A prospective study was undertaken to assess the level of first aid and basic life support (BLS) competence of junior doctors at the Radboud University Nijmegen Medical Centre (RUNMC), the Netherlands. Fifty-four junior doctors (18%), of the medical students in their final years, were submitted to a theoretical test, composed of multiple-choice questions concerning first aid and basic life support. This test was followed by a practical test consisting of two out-of-hospital first aid and basic life support scenarios including cardiopulmonary resuscitation (CPR). In total, 19% of the junior doctors passed the theoretical test. The first scenario was performed correctly in 11%. The CPR situation was correctly performed by 30% of the students as observed by the examiners but when assessed by the checklists of Berden only 6% of the students performed correct CPR. It is concluded that the level of first aid and basic life support of the junior doctors at the RUNMC is low and does not meet the required level as stated in the guidelines for practice of medical education in the Netherlands. PMID- 16707306 TI - EBM user and practitioner models for graduate medical education: what do residents prefer? AB - The objective of the study reported in this article was to assess and explain medical residents' preferences for the evidence based medicine (EBM) practitioner versus the EBM user models. A self-administered survey and focus group of residents attending a core curriculum EBM master session were undertaken. Most residents, particularly those earlier in their training, preferred the practitioner model. Residents perceived that model as an opportunity to gain advanced EBM skills during residency, as providing the ability to choose practicing under both models, and as offering the gain of independent thinking and greater self-confidence in their critical appraisal skills. The user model had the advantage of reduced time requirements. In sum, the majority of residents preferred a curriculum that focuses on the practitioner over the user model. PMID- 16707308 TI - Attitudes to undergraduate medical training. PMID- 16707313 TI - XX European Congress of Perinatal and Neonatal Medicine, Praque, Czech Republic, May 24-27, 2006, Abstracts. PMID- 16707314 TI - What are the instrumentation requirements for measuring the isotopic composition of net ecosystem exchange of CO2 using eddy covariance methods? AB - Better quantification of isotope ratios of atmosphere-ecosystem exchange of CO2 could substantially improve our ability to probe underlying physiological and ecological mechanisms controlling ecosystem carbon exchange, but the ability to make long-term continuous measurements of isotope ratios of exchange fluxes has been limited by measurement difficulties. In particular, direct eddy covariance methods have not yet been used for measuring the isotopic composition of ecosystem fluxes. In this article, we explore the feasibility of such measurements by (a) proposing a general criterion for judging whether a sensor's performance is sufficient for making such measurements (the criterion is met when the contribution of sensor error to the flux measurement error is comparable to or less than the contribution of meteorological noise inherently associated with turbulence flux measurements); (b) using data-based numerical simulations to quantify the level of sensor precision and stability required to meet this criterion for making direct eddy covariance measurements of the 13C/12C ratio of CO2 fluxes above a specific ecosystem (a mid-latitude temperate forest in central Massachusetts, USA); (c) testing whether the performance of a new sensor-a prototype pulsed quantum cascade laser (QCL) based isotope-ratio absorption spectrometer (and plausible improvements thereon)-is sufficient for meeting the criterion in this ecosystem. We found that the error contribution from a prototype sensor (approximately 0.2 per thousand, 1 SD of 10 s integrations) to total isoflux measurement error was comparable to (1.5 to 2x) the irreducible 'meteorological' noise inherently associated with turbulent flux measurements above this ecosystem (daytime measurement error SD of approximately 60% of flux versus meteorological noise of 30-40% for instantaneous half-hour fluxes). Our analysis also shows that plausible instrument improvements (increase of sensor precision to approximately 0.1 per thousand, 1 SD of 10 s integrations, and increased sensor stability during the half-hour needed to integrate eddy covariance measurements) should decrease the contribution of sensor error to the point where it is less than the contribution from meteorological noise. This suggests that new sensors using QCL-based isotope ratio absorption spectroscopy should make continuous long-term observations of the isotopic composition of CO2 fluxes via eddy covariance methods feasible. PMID- 16707315 TI - Isotopic discrimination during litter decomposition and delta13C and delta15N soil profiles in a young artificial stand and in an old floodplain forest. AB - In the present study, rates of litter decomposition and microbial biomass nitrogen were monitored over an 8-month period in a young broadleaf plantation (18 y) and in an old floodplain forest. Moreover, delta13C and delta15N temporal variations within soil profiles were evaluated at both sites. Rates of litter decomposition were higher in spring and autumn than in summer, in both forests. At the end of the observation period the percentage of original litter remaining was not statistically different between the young and the old forest and accounted for 60-70% of the original amount. Microbial biomass nitrogen in the remaining litter and the percentage of litter mass lost during decomposition were positively correlated. The difference in litter quality affected the decomposition rate and also the changes in carbon isotopic composition during the decomposition process. In contrast, 15N isotopic signatures showed a similar trend in the litter of the two forests irrespective of the litter quality. Although delta13Csoil and delta15Nsoil showed considerable temporal variation they increased with depth in the soils of both sites but their seasonal changes did not reflect those of the decomposing litter. Within the same soil horizon, both delta13C and delta15N showed similar seasonal trends in the soils of the two forests, suggesting the involvement of environmental factors acting at regional level, such as soil temperature and rainfall variations, in regulating seasonal delta13C and delta15N soil variations. PMID- 16707317 TI - Flooding of lignite mines: isotope variations and processes in a system influenced by saline groundwater. AB - The quality of both groundwaters and surface waters that arise during flooding of abandoned lignite open pits are influenced by regional and local factors. A typical regional factor is due to oxidised sedimentary sulfides. A more local factor is the interaction of shallow water with highly saline groundwater, which is important in Merseburg-Ost (Germany). Investigation of this system is aided by the use of many environmental isotope tracers but special problems can arise. In order to reveal processes in the mine environment (shallow groundwater, lake water) and to characterise mixtures with saline groundwater results are described using the tracers deltaD, delta18O, delta13C, delta34S, 87Sr/86Sr, 3H, 14C, 39Ar, and 222Rn. Deep highly saline groundwater had a radiocarbon concentration typically below 10 pMC. The values of delta13C(DIC) are around-5 per thousand. As delta13C of the aquifer rock samples (Permian, Zechstein carbonates) was in the range of-6...+5 per thousand, residence time corrections based on delta13C are questionable. Additional checks with 39Ar, as well as results from the variationof delta18O (or deltaD) with respect to the salinity, emphasise a Holocene age; as is also the case for most mineralised groundwaters and also for water having a low delta18O (and deltaD). For saline groundwater residing in the Zechstein aquifer the measured delta34S values of about 12 per thousand are close to those expected from the literature. In contrast, the 87Sr/86Sr ratio of dissolved strontium is far from the values anticipated for the aquifer rocks despite there being proportionality between the chloride concentration and the strontium concentration. Furthermore, the proportionality is not valid in lower mineralised water. The 87Sr/86Sr ratio can, therefore, hardly be used as a tracer for the distribution of ascending saline water. The amount of salt-water coming from below into the residual quarry basins is an essential contribution to the lake inventories. Therefore, 222Rn was used to assist in determining the renewal of salt-water layers that formed in deep lake locations. In the deep zones 222Rn concentrations up to 6 Bq/l were measured but were dominantly in equilibrium with 226Ra, which was found in all higher mineralised groundwater samples. Excess radon was limited to just a few decimetres above the lake sediment surface but does not appear to be caused by continuous groundwater discharge. Hydrochemical investigations of groundwater from the Quaternary aquifer were carried out over the last six years before flooding was complete. Apart from a slight downward shift of the average sulfate concentration, other changes showed virtually no trends. An increase of the sulfate concentration was mostly correlated with a decrease of delta34S for individual sites only, but not for the whole ensemble of sampling locations. Sulfate from pyrite oxidation plays an important role but cannot be attributed unequivocally to coal mining. There are hints that the conditions closer to the basin edges may differ from those remoter parts of the flood plain. PMID- 16707316 TI - Paleodiet characterisation of an Etrurian population of Pontecagnano (Italy) by Isotope Ratio Mass Spectrometry (IRMS) and Atomic Absorption Spectrometry (AAS)(#). AB - Human bones recovered from the archaeological site of Pontecagnano (Salerno, Italy) have been studied to reconstruct the diet of an Etrurian population. Two different areas were investigated, named Library and Sant' Antonio, with a total of 44 tombs containing human skeletal remains, ranging in age from the 8th to the 3rd century B.C. This time span was confirmed by 14C dating obtained using Accelerator Mass Spectrometry (AMS) on one bone sample from each site. Atomic Absorption Spectrometry (AAS) was used to extract information about the concentration of Sr, Zn, Ca elements in the bone inorganic fraction, whilst stable isotope ratio measurements (IRMS) were carried out on bone collagen to obtain the delta13C and delta15N. A reliable technique has been used to extract and separate the inorganic and organic fractions of the bone remains. Both IRMS and AAS results suggest a mixed diet including C3 plant food and herbivore animals, consistent with archaeological indications. PMID- 16707318 TI - Measurement of 137Cs in cultivated soils from two loess areas in Poland. AB - In this paper, a vertical distribution of 137Cs in agricultural soil was investigated. 137Cs is an anthropogenic radioisotope which is strongly adsorbed in soil and can be used to study soil erosion. This paper presents some preliminary results of measurements of activity of 137Cs in soil in Biala and Odonow areas. In both areas, the upper part of the soil profile was cultivated. The experimental results show that the vertical distribution of 137Cs in agricultural soil is different from the vertical distribution of 137Cs in undisturbed soil. The 137Cs is more uniformly distributed in the cultivated soil layer than in an undisturbed site. Migration of 137Cs below plough depth is limited. These data show that the depth distribution of 137Cs in soil profiles for cultivated fields should be considered for precise determination of soil erosion. PMID- 16707319 TI - Radiogenic argon distribution within a mineral grain: implications for dating of hydrothermal mineral-forming event in Sludyanka complex, Siberia, Russia. AB - Usually, in age-determination procedures, the experimenter is interested in the mean concentrations of parent and daughter substances. However, the structure of a solution of the Fick's diffusion equation enables one to specify a procedure, allowing, through macroscopic experiments, one to determine the distribution of radiogenic argon (as well as any of other isotopes) within a mineral grain. Argon, influenced in the geological past by a metamorphic event, has a half sinusoidal distribution, whereas argon preserved in geological time has a rectangular distribution. The ratio of the quantity of argon of the first type to the total argon is called an alpha-factor. It is believed that this coefficient can be of great interest for geochronological studies, being a symptom of virtual Ar losses and the time instant, at which they happened. A laboratory procedure and a mathematical algorithm to determine the alpha-factor are developed. As an example, a determination of the alpha-factor on a vein-phlogopite from the well studied Sludyanka deposit, Southern Baikal, Siberia, Russia, is described. Rb-Sr isochron age on phlogopite-calcite-apatite paragenetic assemblage yielded 460+/-7 Ma and is constrained by U-Pb zircon ages of 471+/-1 and 447+/-2 Ma, respectively, on an emplacement of early syenites and monzonites and later 'post phlogopitic' pegmatites [E.B. Salnikova, S.A. Sergeev, A.B. Kotov, S.Z. Yakovleva, R.H. Steiger, L.Z. Reznitskiy, E.P. Vasil'ev. U-Pb zircon dating of granulite metamorphism in the Sludyanskiy complex, Eastern Siberia. Gondwana Res., 1, 195-205 (1998). L.Z. Reznitskii, G.P. Sandimirova, Y.A. Pakhol'chenko, S.V. Kuznetsova. The Rb-Sr age of phlogopite deposits in Sludyanka, southern Baikal region. Dokl. Earth Sci., 367, 711-713 (1999). L.Z. Reznitskii, A.B. Kotov, E.B. Salnikova, E.P. Vasil'ev, S.Z. Yakovleva, V.P. Kovach, A.M. Fedoseenko. The age and time span of the origin of phlogopite and lazurite deposits in the Southwestern Baikal area: U-Pb geochronology. Petrology, 8, 66-76 (2000).]. In this study, the phlogopite gave an apparent K-Ar age of 365.6+/-10.4 Ma which has no geological meaning. The alpha-factor of 0.27 was derived from a laboratory kinetic experiment. The age determined by the rectangularly distributed argon corresponded to alphax0.129 nmm3/g (total radiogenic argon in the sample)=94.17 nmm3/g, and an age of approximately 274 Ma (e.g. a period without any argon losses). Here, at the first sight, we deal with common argon losses not conjugated with any geological event, because after 447 Ma only lower temperature (in a range of 50-300 degrees C) hydrothermal processes were recorded. A paragenetic phlogopite-bearing vein mineral, hyalophane, having a disturbed Rb-Sr isotopic system, however, yielded 40Ar/39Ar subplateau age of 271+/-2 Ma [V.V. Ivanenko, M.I. Karpenko, M.A. Litsarev. Age of the Sludyanka phlogopite deposits (in Russian; data of the 39Ar-40Ar method). In Geological Series 5, pp. 92-98, Izvestiya Akademii Nauk SSSR, Moskva (1990).], marking similar timing of the K-Ar isotopic system closure. This indicates that K-Ar isotopic system in the phlogopite was not closed (or was partially reset) long after its crystallisation and closing of the Rb-Sr system, in spite of the fact that the crystal was not affected by processes having temperatures above the accepted closure temperatures for micas. There seems to have acted an exchange mechanism of OH-groups between surrounding fluid and hydrocrystal structure. Thus the argon loss did not occur, spontaneously, but argon particles seem to have been captured and carried to the mineral surface by vacancies formed during displacements of OH-groups. PMID- 16707323 TI - Absorption, distribution, and excretion of ammonium perfluorooctanoate (APFO) after oral administration to various species. AB - Male and female mice, rats, hamsters, and rabbits were treated with a single oral dose of 14C-ammonium perfluorooctanoate (APFO), and the excretion and tissue distributions were followed for 120 h (168 h in the rabbit). Substantial sex and species differences in the excretion and disposition of 14C-radioactivity derived from 14C-labeled APFO were observed in this study. The female rat and the male hamster excreted more than 99% of the original 14C activity by 120 h after dosing; conversely, the male rat and the female hamster excreted only 39% and 60% of the original 14C activity, respectively, by 120 h postdosing. The male and female rabbits excreted the 14C activity as rapidly and completely as the female rat and the male hamster, whereas male and female mice excreted only 21% of the original 14C activity by 120 h postdosing. The rapid excretors (female rat, male hamster, and male and female rabbits) contained negligible amounts of 14C in organs and tissues at sacrifice. The slow excretors exhibited the highest 14C concentrations in the blood and liver followed by the kidneys, lungs, and skin. PMID- 16707325 TI - Neurotoxicity evaluation of the organofluorine pesticide etoxazole in the brain of Oreochromis niloticus. AB - Etoxazole is a new organofluorine pesticide that has been used worldwide as acaricide and insecticide since 1998. Almost no previous attempt has been made to evaluate the toxic effects of etoxazole in vertebrates. Using fish (Oreochromis niloticus) as a suitable model organism, the aim of this study was to indicate whether etoxazole affects acetylcholinesterase and sodium potassium-activated adenosine triphosphatase activities in the brain tissue in order to evaluate the impacts on neurotoxicity and ion transportation. Enzyme activities were determined using spectrophotometric methods. At the sublethal concentrations (0.27, 0.54, 0.81, 1.08, 1.35 mg/L) and exposure durations (1, 7, 15 days) tested, etoxazole has no inhibitory effect on the brain acetylcholinesterase and sodium potassium-activated adenosine triphosphatase activities. Our results suggest that etoxazole and/or its metabolites may not reach or penetrate the blood-brain barrier; therefore, they do not essentially alter the functions of these two important enzymes for the brain. PMID- 16707324 TI - Estimation of apoptosis and necrosis caused by pesticides in vitro on human lymphocytes using DNA diffusion assay. AB - Organophosphorus pesticides like monocrotophos, profenofos, chlorpyrifos, and acephate are most commonly used in India for agriculture and public health programs. Previous studies have revealed that at low doses, organophosphorus pesticides not only act as genotoxic agents but also affect several other biochemical pathways. The aim of the current investigation was to assess apoptosis and necrosis caused by these pesticides on human peripheral blood lymphocytes under in vitro conditions using the DNA diffusion assay. Our studies have revealed that all the above pesticides induced apoptosis and necrosis in cultured human peripheral blood lymphocytes in in vitro conditions. The results are statistically significant (p < 0.001). Data on these alterations of immune cells are required for understanding the subchronic effects mediated by pesticides on nontarget organisms. PMID- 16707326 TI - Effects of aspirin and mefenamic acid on soman poisoning-induced neuropathology in mice. AB - The efficacy of aspirin and mefenamic acid to counteract soman-induced brain damage was investigated in mice. Neuronal damage was evaluated in the hippocampus and amygdala by performing omega3 receptor density measurements and hemalun phloxin staining. The effect of both drugs on the proliferation of neural progenitors after soman exposure was also assessed. Mefenamic acid aggravated the soman-induced hippocampal neuropathology. On the other hand, aspirin recorded a weak neuroprotective effect in the amygdala. However, this drug also diminished the proliferation of neural precursor cells. The possible neurochemical mechanisms underlying such differences in the efficacy of the two drugs are also reviewed. PMID- 16707328 TI - Different results of the Salmonella umu test between three isomers of phenylenediamine (PDA) derivatives. AB - A post-treatment assay of the umu test was performed to detect genotoxicity of 10 phenylenediamine (PDA) derivatives using Salmonella typhimurium TA1535/pSK1002 with/without S9 mix. Seven chemicals (o-PDA, 4-chloro-o-PDA, 4-nitro-o-PDA, p PDA, 2-chloro-p-PDA, 2-nitro-p-PDA, and 2,5-diaminotoluene) showed positive results with S9 mix, but three chemicals (m-PDA, 4-chloro-m-PDA, and 2,4 diaminotoluene) were negative with and without S9 mix. Four of 7 chemicals (o PDA, 4-chloro-o-PDA, 4-nitro-o-PDA, and 2-nitro-p-PDA) that gave positive results with S9 mix were also positive without S9 mix. These results indicate that the genotoxicity of PDA derivative possessing m-position amino substituents was not detected in the umu post-treatment assay using TA1535/pSK1002. PMID- 16707327 TI - The first preclinical pharmacotoxicological safety assessment of CGS 35601, a triple vasopeptidase inhibitor, in chronically instrumented, conscious, and unrestrained spontaneously hypertensive rats. AB - CGS 35601 is a triple vasopeptidase inhibitor (VPI) of angiotensin-converting enzyme, neutral endopeptidase, and endothelin-converting enzyme-1 with respective IC50 values of 22, 2, and 55 nM. We characterized the safety profile and toxicity of escalating doses of CGS 35601 over a 20-day period in chronically instrumented, unrestrained, conscious, male, spontaneously hypertensive rats (SHR). Once instrumented with an arterial catheter, the SHR were placed in metabolic cages allowing daily assessment of hemodynamics and blood sampling for biochemical and hematological measurements. After a 7-day stabilization period, the SHR were divided into 2 groups: Gr. 1, (n = 13 to 18) receiving CGS 35601 at 0.01, 0.1, 1 and 5 mg kg(-1) day(-1) (continuous i.a. infusion) for 5 consecutive days/dose, followed by a 5-day washout; and Gr. 2, (n = 10) receiving vehicle (saline). The highest dose of CGS 35601 dose-dependently reduced MABP from 156 +/ 4 up to 94 +/- 5 mm Hg, whereas heart rate, metabolic, electrolytic, and hematological profiles, growth, diuresis, and renal activity were unaffected, and no hepatic or liver toxicities were observed. These results suggest that this novel triple VPI presents no safety concerns at this stage and may become of interest for the treatment of hypertension and other cardiovascular disorders. Long-term chronic experiments are needed to assess possible angioedema and increases in vascular permeability. PMID- 16707329 TI - Evaluation of developmental toxicity of ultraviolet absorber 2-(3',5'-di-tert butyl-2'-hydroxyphenyl)-5-chlorobenzotriazole in rats. AB - 2-(3',5'-Di-tert-butyl-2'-hydroxyphenyl)-5-chlorobenzotriazole (DBHCB) is widely used as a UV absorber. In this study, the developmental toxicity of DBHCB was evaluated in rats. Pregnant rats were given DBHCB at 0, 62.5, 250, or 1000 mg kg( 1) day(-1) by gavage on days 5-19 of pregnancy. No deaths were observed in the pregnant rats of any group. No effect of DBHCB on the general conditions, body weight gain, or feed consumption was observed in the pregnant rats. There were no changes in the ovarian weight, gravid uterine weight, or necropsy findings in the maternal rats of the DBHCB-treated groups. No significant effects of DBHCB were found in the number of corpora lutea, implantations, live fetuses, resorptions or dead fetuses, incidence of pre- or postimplantation embryonic loss, viability of fetuses, fetal weight, or sex ratio of live fetuses. No significant difference in the incidence of fetuses with malformations or variations or degree of ossification was detected between the DBHCB-treated and control groups. PMID- 16707330 TI - The assessment of genotoxicity of carbamazepine using cytokinesis-block (CB) micronucleus assay in cultured human blood lymphocytes. AB - The genotoxic effect of CBZ has been investigated in few studies. There is little evidence linking carbamazepine (CBZ) with any genotoxic effects, particularly in vitro micronucleus test using cytogenesis-block technique. In this study, the genotoxicity of the antiepileptic drug, carbamazepine, was tested using cytokinesis-block (CB) micronucleus assay. In vitro analysis was performed in human blood lymphocytes from four healthy persons at five different concentrations of carbamazepine (6, 8, 10, 12, 14 microg/mL). Genotoxic potential and cytotoxic effects of carbamazepine were evaluated by using micronucleus assay and cytokinesis-block proliferation index (CBPI), called the parameter of cytotoxicity in human peripheral blood lymphocyte cultures, respectively. The results of this study indicate that CBZ caused the genotoxic effect under in vitro conditions, except at the dose of 6 microg/mL, and cytotoxic effects of carbamazepine were revealed by a decrease in the cytokinesis-block proliferation index at all the concentrations. PMID- 16707331 TI - Chemical cross-linking immobilized concanavalin A for use in proteomic analyses. AB - Lectin affinity chromatography was used to reduce the amount of the abundant glycoprotein beta-conglycinin in total protein samples prepared from developing soybean (Glycine max L. Merrill cv. Jack) seeds. Electrophoretic analysis of both the concanavalin A-Sepharose binding and non-binding fraction revealed an abundant protein band at Mr 26,000. The amount of this protein was greatly increased when concanavalin A-Sepharose was used with urea-containing buffers. Peptide mass fingerprint analysis of this abundant protein band unequivocally identified it as concanavalin A (con A). A simple and gentle method was used to chemically cross-link the con A subunits so that the lectin-Sepharose retained the ability to bind high-mannose type glycoproteins. The chemically cross-linked con A-Sepharose was stable in buffers that contained up to 8M urea, making this an affinity matrix suitable for use in electrophoresis-based proteomic analyses. PMID- 16707332 TI - Dyeing of wool fibres with natural dyes: effect of proteolytic enzymes. AB - In spite of the widespread use of proteins (casein, peptone, etc.) and protein fragments as a substrate for the proteolytic enzymes, a substrate prepared from dyes that adsorb onto appropriate materials, such as wool and cotton, are also used for enzyme activity determination. In the point of view of this thought, it was our aim to develop the substrates which are easily and economically obtainable and also environmentally safer for the frequently used proteolytic enzymes, such as subtilisin carlsberg, trypsin, chymotrypsin, and protease type XVI and, if possible, to prepare the specific substrate at least for one of these enzymes. For this aim, wool was dyed with natural dyes such as juglone, lawsone, berberine, and quercetin. The optimum pH, incubation time, and agitation rate were determinated. The results indicate that, of all the tested enzymes on wool dye complex as an insoluble substrate, the most appropriate complex was found to be wool-lawsone complex. PMID- 16707333 TI - Effect of inducers on the production of 5-aminolevulinic acid by recombinant Escherichia coli. AB - Aminolevulinic acid (ALA) was produced by recombinant Escherichia coli BL21(DE3) (pET28-A.R-hemA) harboring the ALA synthase gene (hemA) from Agrobacterium radiobacter zju-0121. The effects of inducers on the ALA synthase activity and ALA productivity were evaluated. The results indicated that a low isopropyl-beta D-thiogalactoside (IPTG) concentration (0.05 mmol/L) was favorable for high expression of ALA synthase, which resulted in higher ALA productivity. For metabolic engineering applications, lactose was a better substitute of IPTG for active enzyme expression. When lactose concentration was 5 mmol/L, the specific ALA synthase activity and ALA productivity reached 16.7 nmol/(min . mg of protein) and 1.15 g/L, respectively, which were about 15% and 43% higher than those induced by IPTG. PMID- 16707334 TI - Preparation of optically pure tert-leucine by penicillin G acylase-catalyzed resolution. AB - Penicillin G acylase, from Kluyvera citrophila, was used in kinetic resolution of DL-tert-leucine. N-phenylacetylated-DL-tert-leucine, chemically synthesized from DL-tert-leucine, was enantioselctively hydrolyzed by penicillin G acylase to obtain L-tert-leucine, D-tert-leucine was prepared by acid-catalyzed hydrolysis of the remaining substrate. The total yields of D-tert-leucine and L-tert-leucine are 80.6% and 83.1%, respectively. The enantiomeric excess of the two products, D tert-leucine and L-tert-leucine, are 98.5% and 99%. This is a practical way for the preparation of D-tert-leucine and L-tert-leucine. PMID- 16707336 TI - Direct haplotype-specific DNA sequencing. AB - Determining haplotype-specific DNA sequence information is very important in a wide range of research fields. However, no simple and robust approaches are currently available for determining haplotype-specific sequence information. We have addressed this problem by developing a very simple and robust haplotype specific sequencing approach. We utilise the fact that DNA sequencing polymerases are sensitive to 3'end mismatches in the sequencing primer. By using two sequencing primers with 3'end corresponding to the two alleles in a given SNP locus, we are able to obtain allele-specific DNA sequences from both alleles. We evaluated this direct haplotype-specific approach by determining haplotypes within the intron 2 sequence of the fructan-6-fructosyltransferase (6-ft) gene in Lolium perenne L. We obtained reliable haplotype-specific sequences for all primers and genotypes evaluated. We conclude that the haplotype-specific sequencing is robust, and that the approach has a potentially very wide application range for any diploid organism. PMID- 16707335 TI - New synthetic route for RGD tripeptide. AB - A new route was employed to synthesize RGD. First, Gly-Asp dipeptide was synthesized by a novel chemical method in two steps, including chloroacetylation of L-aspartic acid and ammonolysis of chloroacetyl L-aspartic acid. Second, Nalpha-Z- L-Arginine was reacted with Gly-Asp to synthesize RGD by the N carboxyanhydride method. Less protected amino acids were used in this synthesis. This method possessed advantages of low cost, simplicity, and rapidity with a reasonable yield of 62% calculated from arginine. In addition, compared with the above method, a conventional solid phase method was also used to synthesize RGD, the yield was 75% calculated from the first amino acid anchored to resin. PMID- 16707337 TI - Immobilization of chitosan-invertase neoglycoconjugate on carboxymethylcellulose modified chitin. AB - Saccharomyces cerevisiae invertase, chemically modified with chitosan, was immobilized on a carboxymethylcellulose-coated chitin support via polyelectrolyte complex formation. The yield of immobilized protein was determined to be 72% and the enzyme retained 68% of the initial invertase activity. The optimum temperature for invertase was increased by 5 degrees C and its thermostability was enhanced by about 9 degrees C after immobilization. The immobilized enzyme was stable against incubation in high ionic strength solutions and was 12.6-fold more resistant to thermal treatment at 65 degrees C than the native counterpart. The prepared biocatalyst retained 98% and 100% of the original catalytic activity after 10 cycles of reuse and 70 h of continuous operational regime in a packed bed reactor, respectively. The immobilized enzyme retained 95% of its activity after 50 days of storage at 37 degrees C. PMID- 16707338 TI - Expression and characterization of earthworm Eisenia foetida lumbrokinase-3 in Pichia pastoris. AB - Lumbrokinase-3 (LK-3, AY438622), first cloned from the earthworm Eisenia foetida in our laboratory, is a component of earthworm fibrinolytic enzymes. In this study, cDNA encoding the LK-3 gene was sub-cloned into yeast pPIC9K expression vector and transformed into the Pichia pastoris GS115 cells by electroporation. High level expression of LK-3 in yeast cells was confirmed with a different induction time. The activity of expressed LK-3 was observed in fibrin plates. In addition, the expressed LK-3 protein could dissolve fibrinogen and bovine serum albumin. The use of this system for the high level production of biological protein is implicated from this study. PMID- 16707341 TI - Methodological issues in comparing injury incidence across countries. AB - The primary objective was to describe the methodological challenges and devise solutions to compare injury incidence across countries. The research design was a mixed methods study, consisting of a consultation with an expert group and comparison of injury surveillance systems and data from ten European countries. A subset of fractures, selected radiologically verifiable fractures and a method of checking the national representativeness of sample emergency department data were devised and are proposed for further development. These methodological considerations and developments will be further refined and tested and should prove useful tools for those who need to compare injury incidence data across countries. PMID- 16707342 TI - Motorcycle safety programmes in Malaysia: how effective are they? AB - This paper presents the approach taken by the Malaysian Government to contain motorcycle casualties in Malaysia. It involves the exposure control, crash prevention, behaviour modification and injury control related to humans, vehicles and the environment based on pre-crash, crash and post-crash scenarios of motorcycle accidents. These initiatives emanated mainly from the research and development carried out by the Road Safety Research Centre at Universiti Putra Malaysia. Recent outcomes from these initiatives are presented and their impact is highlighted. PMID- 16707343 TI - Alcohol as a risk factor for unintentional rail injury fatalities during daylight hours. AB - Railway fatalities account for approximately 10% of transport fatalities in Cape Town. The objective of this study was to examine alcohol intoxication as a risk factor during daylight hours by conducting a case - control study to compare rail passenger and pedestrian fatalities (cases) with motor vehicle passenger and pedestrian fatalities (controls). Rail passenger and rail pedestrian fatalities were defined as cases with motor vehicle passenger and pedestrian fatalities as the respective controls. Data were collected from post-mortem reports at two mortuaries from 1994 to 1996. Blood alcohol concentration was the dependent variable. The independent variables were age, gender, date of death, day of week, time of injury and external cause of death. The late afternoon and early evening period from 1600 hours to 1900 hours had the highest frequency of fatalities for all case and control groups. Of the 56 predominately male (89%) railway passenger cases with an average age of 34.5 (SD 12.5) years, Friday (27%) was the most frequent day of death. Railway pedestrian cases (89% male, average age 36.8 years (SD 13.3)) were more likely to be killed on a Monday (11% of cases). Among the controls, motor vehicle passengers (63% male, average age 39.9 (SD 15.5)) were more likely to die on a Sunday (25%) and pedestrians (82% male, average age 41 (SD 14.7)) on a Saturday (21%). The study showed that alcohol consumption is an important risk factor for rail fatalities during daylight hours, with rail passenger fatalities being 4.71 (1.72 - 12.88) and rail pedestrian fatalities 1.62 (0.98 - 2.69) times more likely to be intoxicated than the respective controls. The results provide more evidence for public health campaigners to tackle endemic alcohol abuse and to develop diverse interventions that do not exclusively target motor vehicle drivers. PMID- 16707344 TI - Severity of alcohol-related motor vehicle crashes in British Columbia: case - control study. AB - The objective of the present study was to compare the injury severity and vehicle damage severity rates of alcohol-related crashes with rates of non-alcohol related crashes in British Columbia (BC). Injury severity rates and vehicle damage severity rates were taken from 2002 Insurance Corporation of British Columbia traffic collision data. The data were computed in order to compare the differences in injury severity and vehicle damage severity rates of alcohol related vs. non-alcohol-related motor vehicle crashes. Case - control methods were used in this study to analyse the risk of alcohol-related crashes compared to non-alcohol-related crashes in BC. Odds ratios (OR) and 95% CI were calculated to estimate relative risks. In the case - control analysis, the risk of fatal collision was increased for those drinking and driving compared with those driving sober (OR 4.70; 95% CI 3.15 - 7.01). Risk of injury collision was increased for those drinking and driving compared with those driving sober (OR 1.32; 95% CI 1.19 - 1.37). Importantly, the risk of vehicle damage severity was increased for those drinking and driving compared with those driving sober (write off vehicle OR 4.24; 95% CI 3.70 - 4.86, severely damaged vehicles OR 1.98; 95% CI 1.77 - 2.21). The study reinforces existing literature to suggest that current evidence is sufficient to show an increased risk of injury and fatality to drivers and occupants in alcohol-related crashes. This paper not only emphasizes this well-known relationship, but also such consequences as increased vehicle damage severity. The connection between drinking and severity of motor vehicle crashes is popularly believed and has now received substantial scientific support. There is strong justification for injury prevention experts and policy makers to step up motor vehicle crash injury prevention advocacy by implementing evidence-based policies to reduce rates of alcohol-impaired driving in the province of BC. Most unintentional injuries in BC are related to motor vehicle crashes. Significant improvements can be made in these statistics by: increasing the use of occupant protection (safety belt and child restraint seats); reducing alcohol-related injuries through multiple strategies including corrections in the physical environment, extensive enforcement of drinking and driving laws and health promotion/education. PMID- 16707345 TI - The association of light trucks and vans with paediatric pedestrian deaths. AB - The hypothesis that relative to cars, light trucks and vans (including sports utility vehicles) are more likely to result in fatal paediatric pedestrian injury was investigated. It was further hypothesized that this increased risk is a result of head injuries. The study sample consisted of 18 117 police records of motor vehicles involved in crashes in which one or more pedestrians aged 5 to 19 years old was injured or killed. Frequencies and case fatality ratios for each vehicle body type were calculated. A logistic regression analysis was conducted, with light truck or van vs. car as the exposure variable and fatal/non-fatal pedestrian injury as the outcome variable. After controlling for driver age, driver gender, vehicle weight, road surface condition and presence of head injury, 5 to 19 year-olds struck by light trucks or vans were more than twice as likely to die than those struck by cars (odds ratio (OR) 2.3; 95% CI 1.4, 3.9). For the 5 to 9 year-old age group, light trucks and vans were four times as likely to be associated with fatal injury (OR 4.2; 95% CI 1.9, 9.5). There was an association between head injury and light trucks and vans (OR 1.2; 95% CI 1.1, 1.3). It was concluded that vehicle body type characteristics play an important role in paediatric pedestrian injury severity and may offer engineering-based opportunities for injury control. PMID- 16707346 TI - Drowning--a major but neglected child health problem in rural Bangladesh: implications for low income countries. AB - This study was intended to estimate the magnitude and explore the determinants of childhood drowning in rural Bangladesh. A cross-sectional survey as well as a population-based case - control study was conducted. By multistage cluster sampling 51 147 children aged 1 - 4 years were identified from 108 827 rural households. All drownings in children aged 1 - 4 years in the preceding 5 years were identified and recruited as cases and two living children of the same age group were selected from the same localities as controls. Socio-economic, demographic, environmental and other related information was collected from mothers of both cases and controls by face-to-face interview with the help of structured questionnaires. The incidence of drowning among children aged 1 - 4 years old was 156.4 per 100 000 children-year. The highest rate (328.1 per 100 000; 95% CI 254.8 - 421.7) was observed in 1 year old male children. The proportional mortality due to drowning in the children was 27.9%. Mothers' age and literacy and family income were identified as risk factors. Drowning is one of the major causes of 1 - 4 years childhood mortality in Bangladesh. One-year old male children from poor families were at great risk of drowning in rural Bangladesh. PMID- 16707347 TI - Foreign travel and the risk of harm. AB - Systematic data on the risk of fatal and non-fatal injury from external causes to those who travel abroad for purposes of business or pleasure are seldom recorded and are consequently hard to come by. In this paper, the risk of fatal injuries to foreign travellers using historical and newly acquired data from national databases is estimated. Overall, it appears from these data that the risk of fatal injury to foreign travellers lies in the range of 20 to 90 per 100 000 person-years of exposure for many destinations worldwide, although different rates may pertain in selected situations where special conditions apply, for example, where there is heavy participation in risky sports or increased driving. This level of fatal injury risk is not noticeably different from that of staying in one's home country if one lives in a Western-style industrial country, although the types of hazards responsible may vary in type and proportion. With increasing levels of travel, and the growing popularity of more adventurous pursuits, it is possible that risks could be increasing in this sector. PMID- 16707348 TI - Prognosis of trauma patients admitted to an emergency department in a developing country. PMID- 16707349 TI - A survey of inventions aimed at preventing drowning. PMID- 16707350 TI - Not all risk is bad, playgrounds as a learning environment for children. PMID- 16707351 TI - Improvements in trauma care capabilities in Vietnam through use of the WHO-IATSIC Guidelines for Essential Trauma Care. PMID- 16707352 TI - Pattern of road traffic injuries: one year hospital-based study in Western Nepal. AB - Road traffic injuries and deaths caused by motor vehicles is a growing public health problem all over the world. Inter-country or regional differences in the pattern of injury by road users have significant implications in determining prevention policies. The present study was conducted to evaluate the pattern of injuries in 217 hospitalized trauma patients admitted during 1 year, 2003. The majority of the injuries (54%) involved motorcycles. The highest incidence amongst the male population was in the age group of 16 - 30 years. Head and face injuries and injuries to the lower limbs comprised 58.1% and 50.7% of all injuries respectively. The bones of the lower limbs were most commonly fractured. To conclude, the traffic casualties of motorcyclists and pedestrians are considered a major problem and the preventive measures to reduce these transport related injuries are discussed in this study. PMID- 16707354 TI - Animal models of drug addiction: Where do we go from here? AB - Compulsion and impulsivity are both primary features of drug addiction. Based on decades of animal research, we have a detailed understanding of the factors (both environmental and physiological) that influence compulsive drug use, but still know relatively little about the impulsive aspects of drug addiction. This review outlines our current knowledge of the relationship between impulsivity and drug addiction, focusing on cognitive and motor impulsivity, which are particularly relevant to this disorder. Topics to be discussed include the influence of chronic drug administration on impulsivity, the mechanisms that may explain drug induced impulsivity, and the role of individual differences in the development of impulsive drug use. In addition, the manner in which contemporary theories of drug addiction conceptualize the relationship between impulsivity and compulsion is examined. Most importantly, this review emphasizes a critical role for animal research in understanding the role of impulsivity in the development and maintenance of drug addiction. PMID- 16707355 TI - Induction when rats lick for 1% liquid-sucrose reinforcement. AB - Previous research reported that rats responding for 1% liquid-sucrose reinforcement when 32% sucrose reinforcement is upcoming will decrease their response rate (contrast) if licking is the dependent measure and increase their response rate (induction) if pressing a lever is the dependent measure. The present study investigated whether induction could be observed when licking served as the dependent measure and whether induction in lever pressing and contrast in licking behaviour could be concurrently observed. Experiment 1 found induction when rats licked to earn the rewards but consumed them at a location separate from the spout licked to earn them. Experiment 2 also found induction when rats earned (and consumed) rewards by licking the same spout throughout the session. Experiment 3 separately measured instrumental lever pressing for sucrose rewards and licking the sucrose during the reward period. We found that both measures increased for 1% sucrose when 32% sucrose reinforcement was upcoming. The present results indicate that the type of response is not the sole determinant of whether contrast or induction is observed. Rather, they suggest that other procedural details, such as the location of reinforcer delivery, influence which effect is observed. The results also indicate that associative processes underlie the appearance of induction in responding for 1% sucrose. PMID- 16707356 TI - Classification and recognition in artificial grammar learning: Analysis of receiver operating characteristics. AB - In two experiments we investigated recognition and classification judgements using an artificial grammar learning paradigm. In Experiment 1, when only new test items had to be judged, analysis of z-transformed receiver operating characteristics (z-ROCs) revealed no differences between classification and recognition. In Experiment 2, where we included old test items, z-ROCs in the two tasks differed, suggesting that judgements relied on different types of information. The results are interpreted in terms of heuristics that people use when making classification and recognition judgements. PMID- 16707357 TI - Negative priming is stronger for task-relevant dimensions: Evidence of flexibility in the selective ignoring of distractor information. AB - The finding that in selective attention tasks responses to previously ignored stimuli are usually retarded is known as negative priming (NP). In previous studies it has been suggested that NP can depend on behavioural goals--that is, NP was observed only for task-relevant object dimensions. We extend these findings with two experiments demonstrating that stronger NP can be observed for task-relevant dimensions than for task-irrelevant dimensions (a) even if participants' tasks vary blockwise within an experiment and (b) if behavioural goals vary from trial to trial. These results suggest that selective NP is a much more flexible process than previously assumed. PMID- 16707358 TI - On the locus of dual-task interference: Is there a bottleneck at the stimulus classification stage? AB - Recent studies have provided evidence that dual-task interference is typically caused by a single-channel bottleneck, but the processing locus of the bottleneck has yet to be pinned down. A bottleneck locus at the response-selection stage is widely advocated, but an earlier locus would be consistent with most previous evidence. Four new experiments used the "locus of slack" method to investigate whether the stages postponed by the central bottleneck include stimulus classification, a very late stage of perceptual processing. The experiments varied stimulus classification difficulty for two different analogueue perceptual judgements. Experiment 1 found only modest absorption into slack for the difficulty of a spatial position judgement. Experiments 2-4 found virtually no absorption into slack for the difficulty of a box-width judgement. These results support a bottleneck locus beginning at or before the stage of stimulus classification and hence prior to the stage of response selection. Other evidence, however, leaves no doubt that response selection is also subject to bottleneck postponement. Two architectures are discussed that can account parsimoniously for both old and new results. One posits a single bottleneck resulting from a unified CPU-like central processor; the other posits multiple bottlenecks resulting from multiple processors accomplishing different substages of central processing. PMID- 16707359 TI - Dual route for subtask order control: Evidence from the psychological refractory paradigm. AB - A change in subtask order in the psychological refractory period (PRP) paradigm increases the effect of stimulus onset asynchrony (SOA) on the second response. We used a paradigm with cued, randomly determined subtask order to test the hypothesis that this SOA by order switch overadditivity reflects order control, via "copying" stimulus order. In Experiments 1a and 1b, overadditivity was evident only with insufficient opportunity for cue-based order control. In Experiment 2, overadditivity was decreased by using the same set of stimuli in the two subtasks, presumably by removing the opportunity to rely on stimulus order. In Experiment 3, removing the order cue increased the overadditivity, presumably because control was based solely upon copying stimulus order. The results indicate interactive top-down and bottom-up order control. Implications to theories of the PRP paradigm are discussed. PMID- 16707360 TI - Executive functions and achievements in school: Shifting, updating, inhibition, and working memory. AB - Links have recently been established between measures of educational attainment and both verbal and visuo-spatial aspects of working memory. Relationships have also been identified between specific executive functions-shifting, updating, and inhibition-and scholastic achievement. In the present study, scholastic attainment, shifting, updating, inhibition, and verbal and visuo-spatial working memory were assessed in 11- and 12-year-old children. Exploratory factor analysis identified two executive factors: one associated with updating functions and one associated with inhibition. Updating abilities were closely linked with performance on both verbal and visuo-spatial working memory span tasks. Working memory was closely linked with attainment in English and mathematics, and inhibition was associated with achievement in English, mathematics, and science. Domain-specific associations existed between verbal working memory and attainment in English, and between visuo-spatial working memory and attainment in English, mathematics and science. Implications of the findings for the theoretical analysis of executive functioning, working memory and children's learning are discussed. PMID- 16707361 TI - Linguistic and nonlinguistic influences on the eyes' landing positions during reading. AB - Two eye tracking experiments show that, for near launch sites, the eyes land nearer to the beginning of words with orthographically irregular than with regular initial letter sequences. In addition, the characteristics of words, at least at the level of orthography, influence the direction and length of within word saccades. Importantly, these effects hold both for lower case and for visually less distinctive upper case text. Furthermore, contrary to previous evidence (Tinker & Paterson, 1939), there is little effect of type case on reading times. Additional analyses of oculomotor behaviour suggest that there is an inverted optimal viewing position for single fixation durations on words. Both the supplementary analyses and the effects of orthography on fixation positions are relevant to current models of eye movements in reading. PMID- 16707362 TI - It looks easy! Heuristics for combinatorial optimization problems. AB - Human performance on instances of computationally intractable optimization problems, such as the travelling salesperson problem (TSP), can be excellent. We have proposed a boundary-following heuristic to account for this finding. We report three experiments with TSPs where the capacity to employ this heuristic was varied. In Experiment 1, participants free to use the heuristic produced solutions significantly closer to optimal than did those prevented from doing so. Experiments 2 and 3 together replicated this finding in larger problems and demonstrated that a potential confound had no effect. In all three experiments, performance was closely matched by a boundary-following model. The results implicate global rather than purely local processes. Humans may have access to simple, perceptually based, heuristics that are suited to some combinatorial optimization tasks. PMID- 16707363 TI - Diagnostic assays for the JAK2 V617F mutation in chronic myeloproliferative disorders. PMID- 16707364 TI - Infectious diseases and pathology: opportunities and challenges. PMID- 16707365 TI - Molecular biogrouping of pathogenic Yersinia enterocolitica: development of a diagnostic PCR assay with histologic correlation. AB - Yersinia enterocolitica (YE) is associated with several inflammatory gastrointestinal disorders. Pathogenic YE organisms are classified as biogroup 1B (high-virulence [HV] serovars) or biogroups 2 through 5 (low virulence [LV]). We developed the first molecular assay designed to distinguish between these groups and correlated the molecular results with histologic patterns of inflammation. Eleven known pathogenic YE culture isolates (6 biogroup 1B and 5 biogroups 2-5) and 6 YE-positive archival cases were subjected to polymerase chain reaction analysis using primer pairs targeting a strain-dependent variable region, allowing discrimination between biogroups with a single assay. All 11 known culture isolates were confirmed. Of the 6 archival cases, 4 were LV, and 2 were HV. Histologic correlation revealed granulomatous inflammation in the LV cases and suppurative inflammation in the HV cases. This novel assay is useful for diagnosis using culture samples and archival tissues. It also could yield important information correlating YE epidemiology, pathogenesis, and morphology because these preliminary data suggest that LV strains may be associated with chronic granulomatous processes and HV strains with suppurative inflammation. PMID- 16707366 TI - Performance characteristics of two real-time PCR assays for the quantification of Epstein-Barr virus DNA. AB - We compared the performance of a laboratory-developed 5'-nuclease real-time polymerase chain reaction assay and a commercial assay (LightCycler, Roche Diagnostics, Indianapolis, IN) for quantification of Epstein-Barr virus (EBV) DNA. Using standards provided by the manufacturer, the LightCycler assay was linear from 100 to 1 million copies per reaction. Based on dilution of a plasmid containing the amplicon, the laboratory-developed assay was linear from 22 to 45 million copies per reaction. Both assays detected 0.5 copies of genomic EBV DNA per reaction; both showed good reproducibility with coefficients of variation from 0.3% to 2.4% for the LightCycler and 1.8% to 5.1% for the laboratory developed assay. For 31 whole blood specimens with measurable values in both assays, the viral load values obtained with the LightCycler averaged 2.3-fold higher than those obtained with the laboratory-developed assay. Testing 30 matched whole blood and plasma samples in the laboratory-developed assay showed whole blood viral load values averaged 10-fold higher than those for plasma. The LightCycler and laboratory-developed assays are sensitive and reproducible with broad linear ranges. Further clinical evaluation is needed to determine the viral load cutoff that is predictive of posttransplantation lymphoproliferative disorders. PMID- 16707367 TI - Prostate-specific antigen, high-molecular-weight cytokeratin (clone 34betaE12), and/or p63: an optimal immunohistochemical panel to distinguish poorly differentiated prostate adenocarcinoma from urothelial carcinoma. AB - An optimal immunohistochemical panel to distinguish poorly differentiated prostate (PCa) from urothelial (UCa) carcinoma was selected from a panel consisting of prostate-specific antigen (PSA) and prostatic acid phosphatase (PAP), high-molecular-weight cytokeratin (HMWCK), clone 34betaE12, cytokeratin (CK) 7, CK20, p63, and alpha-methylacyl-coenzyme A racemase. The pilot group was composed of poorly differentiated UCa (n = 36) and PCa (n = 42). PSA and PAP stained 95% of PCa vs 0% and 11% of UCa cases, respectively. HMWCK and p63 stained 97% and 92% of UCa vs 2% and 0% of PCa cases respectively. CK7/CK20 coexpression was noted in 50% of UCa cases, whereas 86% of PCa cases were negative with both. A panel of PSA, HMWCK, and p63 was optimal for separating 95% PCa (PSA+/HMWCK and/or p63-) vs 97% UCa (PSA-/HMWCK and/or p63+). This panel was used on 26 diagnostically challenging cases and resolved 81% of cases as UCa vs PCa. The majority of PCa cases retain PSA. Negative PSA with positive HMWCK and/or p63 establishes a diagnosis of UCa. PMID- 16707368 TI - Immunohistochemical differential diagnosis between large cell neuroendocrine carcinoma and small cell carcinoma by tissue microarray analysis with a large antibody panel. AB - To elucidate additional phenotypic differences between large cell neuroendocrine carcinoma (LCNEC) and small cell lung carcinoma (SCLC), we performed tissue microarray (TMA) analysis of surgically resected LCNEC and SCLC specimens. Immunostaining with 48 antibodies was scored based on staining intensity and the percentage of cells that stained positively. Four proteins were identified as significantly expressed in LCNEC as compared with SCLC: cytokeratin (CK)7, 113 vs 49 (P < .0301); CK18, 171 vs 60 (P < .0008); E-cadherin, 77 vs 9 (P < .0073); and beta-catenin, 191 vs 120 (P < .0286). Immunostaining of cross-sections containing LCNEC and SCLC components revealed significant expression of CK7, CK18 and beta catenin in the LCNEC component compared with the SCLC component in 2 of 3 cases. Our results indicate that significant expression of CK7, CK18, E-cadherin, and beta-catenin is more characteristic of LCNEC than of SCLC, and these findings provide further support that these tumor types are separate entities morphologically and immunophenotypically, if not biologically. PMID- 16707370 TI - Repeat thyroid nodule fine-needle aspiration in patients with initial benign cytologic results. AB - Fine-needle aspiration (FNA) is the standard of care for the initial workup of thyroid nodules, but there is no consensus algorithm to manage patients with benign results. We examined performance characteristics of initial and repeat satisfactory FNAs for all 402 patients who underwent thyroid surgery during a recent 22-month period. Of these patients, 267 had at least 1 satisfactory FNA and 70 had 2 or more. After an initial benign FNA, 1 repeat FNA correctly identified an unsuspected malignancy in 2 of 70 patients and was indeterminate in 17; of these, 7 of 17 were identified as malignant in the final pathologic diagnosis. Overall, the use of 1 repeat FNA increased the sensitivity for malignancy from 81.7% to 90.4% and decreased the false-negative rate from 17.1% to 11.4%. With more than 1 repeat FNA, there was no improvement in performance characteristics. These data make a strong argument for 1 repeat FNA following an initial benign FNA diagnosis. PMID- 16707369 TI - Reproducibility of MDM2 and CDK4 staining in soft tissue tumors. AB - MDM2 and CDK4 immunostaining can be useful adjuncts in diagnosing liposarcoma among soft tissue neoplasms. We examined the reproducibility of MDM2 and CDK4 staining between 2 laboratories and between tissue microarrays and whole tissue sections. Sixty-two soft tissue tumors were immunostained at the Bergonie Institute, Bordeaux, France, and the Curie Institute, Paris, France. We also examined 203 soft tissue neoplasms on standard tissue sections and tissue microarrays. There was high concordance of results obtained from the 2 laboratories (with 2 different pathologists) for MDM2 (kappa, 0.93) and CDK4 (kappa, 0.8) staining. There also was excellent concordance between results on tissue microarray and on whole tissue sections for MDM2 (kappa, 0.80) and CDK4 (kappa, 0.93). Immunostaining for MDM2 and CDK4 is a reproducible technique that may be exported to different laboratories for routine use. Tissue microarray is indicated for studying large series. PMID- 16707371 TI - Diagnosis of deep-seated lymphoma and leukemia by endoscopic ultrasound-guided fine-needle aspiration biopsy. AB - We retrospectively studied the use of endoscopic ultrasound-guided fine-needle aspiration biopsy (EUS-FNAB) as a tool for the diagnosis of deep-seated lymphoma. An on-site assessment at the time of EUS-FNAB was performed by a cytopathologist using Diff-Quik (American Scientific Products, McGraw Park, IL) stain. In addition, Papanicolaou stains were performed on EUS-FNAB smears, immunohistochemical stains were performed on cell blocks, and additional samples were sent for flow cytometric analysis. Final cytologic diagnosis was correlated with surgical pathology and/or clinical follow-up. We evaluated EUS-FNAB specimens of deep-seated lymph nodes, spleen, stomach, and pancreas, and 1 EUS guided needle core biopsy specimen of a lymph node. Thirteen cases of deep-seated lymphoma were diagnosed, including non-Hodgkin lymphomas and Hodgkin lymphoma. One case of hairy cell leukemia was diagnosed. EUS-FNAB is a minimally invasive, cost-effective, and useful tool for the primary diagnosis or staging of deep seated lymphomas. PMID- 16707372 TI - A systematic review of the quality of liver biopsy specimens. AB - Characteristics for an optimal liver biopsy specimen were recently defined as 20 to 25 mm long and/or containing more than 11 complete portal tracts (CPTs). A systematic review of percutaneous liver biopsy (PLB) and transjugular liver biopsy (TJLB) series yielded only 32 PLB studies in which these characteristics were evaluated: mean +/- SD length, 17.7 +/- 5.8 mm and number of CPTs, 7.5 +/- 3.4; and 15 TJLB studies: mean +/- SD length, 13.5 +/- 4.5 mm and number of CPTs, 6.8 +/- 2.3. Studies of sampling heterogeneity and intraobserver and interobserver variability also used inadequate specimens by present standards. Only 11 (5.3%) of 207 therapeutic studies for chronic hepatitis B and C documented length and/or number of CPTs. Of the current 12 studies evaluating noninvasive fibrosis tests, only 8 documented length or number of CPTs, and only 1 documented length and number of CPTs. New studies are needed based on adequate liver biopsy samples to provide reliable estimation of grading and staging in chronic liver disease. PMID- 16707373 TI - The binding of thyroid transcription factor-1 and hepatocyte paraffin 1 to mitochondrial proteins in hepatocytes: a molecular and immunoelectron microscopic study. AB - We studied the antibodies hepatocyte paraffin 1 (Hep Par 1) and thyroid transcription factor-1 (TTF-1; clone 8G7G3/1) in normal human liver tissue with immunoelectron microscopy using renal tubules as control samples. TTF-1 (8G7G3/1) and Hep Par 1 bound exclusively to the mitochondria of hepatocytes but not to those of the renal tubular epithelium. Both antibodies labeled mitochondria in a similar pattern and density. These findings confirm that the binding site of Hep Par 1 in hepatocytes is mitochondrial. The specific binding of TTF-1 (8G7G3/1) in hepatocyte mitochondria suggests its potential usefulness for identifying hepatocellular carcinoma. Western blot analysis with cellular proteins extracted from normal human liver and thyroid tissue demonstrated that Hep Par 1 and TTF-1 (8G7G3/1) bound to a protein band of approximately 150 kd in liver cells, with TTF-1 (8G7G3/1) showing less affinity. It is likely that different epitopes to Hep Par 1 and TTF-1 (8G7G3/1) share the same protein molecule in hepatocyte mitochondria. PMID- 16707374 TI - Usefulness (or lack thereof) of immunophenotyping in atypical cutaneous T-cell infiltrates. AB - Our purpose was to evaluate the interobserver concordance for the diagnoses of mycosis fungoides (MF), atypical dermatoses (AD), and benign dermatoses (BD) and the impact of T-cell immunophenotyping on the diagnoses MF, AD, and BD. Specimens of MF (n = 57), AD (n = 27), BD and normal skin (n = 54) were reviewed by 2 hematopathologists and 1 dermatopathologist to establish diagnostic interobserver concordance by routine morphologic examination. Immunophenotyping was performed to evaluate expression of CD2, CD3, CD4, CD5, CD7, CD8, CD20, CD30, and MIB-1. The interobserver concordance was fair to moderate compared with the original diagnosis. Partial deletion of CD2 alone was associated significantly with MF. Epidermal deletions of 2 or 3 T-cell antigens or 2 T-cell antigens not including CD7 were associated significantly with MF. An elevated CD4/CD8 ratio correlated with MF. Morphologic features were most diagnostic of MF. Immunophenotyping generally resulted in downgrading of the reaction pattern but was helpful in distinguishing MF from benign dermatoses. PMID- 16707375 TI - Measuring the value of review of pathology material by a second pathologist. AB - In many departments, some cases are reviewed routinely by a second pathologist within the same department before sign out. The value of this practice is not known. We reviewed and compared the disagreement and amendment rates for cases reviewed by 1 or more pathologists based on the results of blinded review. A total of 8,363 cases underwent blinded review, and of these, 1,087 (13.0%) were reviewed by more than 1 pathologist before sign out. The disagreement rate for cases reviewed by more than 1 pathologist (4.8%) was significantly lower than for cases reviewed by only 1 pathologist (6.9%; P = .004). The amendment rate decreased to 0.0% from 0.5%, but this decrease was not statistically significant (P = .12). Review of material by a second pathologist before sign out is associated with a lower disagreement rate. These results suggest second review of surgical pathology is of value, but the best selection of cases to be reviewed remains to be defined. PMID- 16707376 TI - Promoter hypermethylation of a novel gene, ZHX2, in hepatocellular carcinoma. AB - We used methylation-sensitive restriction fingerprinting (MSRF) to identify novel CpG (5'-CG-3' palindrome; p, phosphate group)-rich sequences that are methylated differentially between the hepatocellular carcinoma (HCC) genomes and adjacent nontumorous liver tissues. A 199-base-pair sequence methylated in HCC tumor tissue was isolated and showed high homology to the 5' CpG island of the zinc fingers and homeoboxes protein 2 (ZHX2) gene. By using bisulfite sequencing, we confirmed that hypermethylation of the 5' CpG island of ZHX2 occurred in some HCC and HepG2 cell lines but not in 6 normal liver tissue samples. By using methylation-specific polymerase chain reaction, we detected methylation of the 5' CpG island of ZHX2 in 46.9% of the HCCs. Reverse transcription-polymerase chain reaction demonstrated that ZHX2 messenger RNA (mRNA) was expressed in all 6 normal liver tissue samples but in only 13.3% of the methylated HCCs. Treatment of HepG2 with 5-aza-deoxycytidine could demethylate the promoter and increase ZHX2 mRNA expression. These results suggest that hypermethylation-mediated silencing of ZHX2 is an epigenetic event involved in HCC. PMID- 16707377 TI - The analytic sensitivity and mutant detection capability of six hepatitis B surface antigen assays. AB - Hepatitis B virus surface antigen (HBsAg) mutants occur in clinical specimens. We studied the analytic sensitivity and ability to detect recombinant and native mutants of 6 HBsAg assays. The ARCHITECT, AUSZYME MONOCLONAL and AxSYM assays (Abbott Diagnostics, Abbott Park, IL), the ADVIA Centaur assay (Bayer Diagnostics, Tarrytown, NY), and the Test System 3 and VITROS ECi assays (Ortho Clinical Diagnostics, Raritan, NJ) showed comparable sensitivity with wild-type HBsAg. The ARCHITECT, AUSZYME, and AxSYM assays detected all mutants that were tested. The Test System 3 and VITROS ECi assays failed to detect mutants with amino acid substitutions at positions 143, 144, and 145, which are located in the immunodominant "a" determinant. The ADVIA Centaur failed to detect substitutions at position 145 and showed negative or very low positive results for substitutions at position 143. The inability to detect HBsAg mutants may lead to misdiagnosis of hepatitis B virus infection. Further studies on the prevalence of HBsAg mutants and the ability of commercial assays to detect them are needed. PMID- 16707378 TI - Performance characteristics of seven automated CA 15-3 assays. AB - Measurements of serum cancer antigen (CA) 15-3 are used to monitor tumor recurrence and treatment of advanced disease. We evaluated the performance characteristics, including limit of detection, linearity, method comparison, and reference intervals, of 7 automated methods for CA 15-3, including the Access 2 (Beckman Coulter, Brea, CA), ADVIA Centaur (Bayer Diagnostics, Tarrytown, NY), ARCHITECT i2000 and AxSYM (Abbott Diagnostics, Abbott Park, IL), Elecsys 2010 (Roche Diagnostics, Indianapolis, IN), IMMULITE 2000 (Diagnostic Products, Los Angeles, CA), and VITROS ECi (Ortho Clinical Diagnostics, Raritan, NJ) assays. The limit of detection for each assay was less than 1.0 kU/L. The maximum deviation for the target values for linearity samples was less than 10% for all methods. Method comparison studies revealed large differences for some individual samples. Overall slopes ranged from 0.50 to 1.48, and correlation coefficients were 0.90 to 0.96 when the ADVIA Centaur was the comparison method. The 97.5 percentile upper reference limit ranged from 23.3 to 51.7 kU/L. Additional standardization efforts are needed, and the availability of reference material is required. Substantial intermethod differences exist for some patient samples, indicating that redetermining the baseline is required when changing methods. PMID- 16707379 TI - Analysis of laboratory critical value reporting at a large academic medical center. AB - Reporting of laboratory critical values has become an issue of national attention as illustrated by recent guidelines described in the National Patient Safety Goals of the Joint Commission on Accreditation of Healthcare Organizations. Herein, we report the results of an analysis of 37,503 consecutive laboratory critical values at our institution, a large urban academic medical center. We evaluated critical value reporting by test, laboratory specialty, patient type, clinical care area, time of day, and critical value limits. Factors leading to delays in critical value reporting are identified, and we describe approaches to improving this important operational and patient safety issue. PMID- 16707380 TI - Total laboratory automation can help eliminate the laboratory as a factor in emergency department length of stay. AB - We obtained data on laboratory turnaround time (TAT) and emergency department (ED) length of stay (LOS). We correlated potassium test TAT outlier percentage (TAT-OP) with ED LOS and found that for each outlier percentage (potassium result > 40 minutes), a projected impact on ED LOS was approximately 2.8 additional minutes (ED LOS = 2.79 TAT-OP + 78.77). To address this issue, we began implementation of a totally automated chemistry system to decrease TAT-OPs. Our TAT means did not change substantially with automation (potassium, 28 to 27 minutes); however, TAT-OPs decreased substantially (potassium, 18% to 5%). Preautomation average ED LOS correlated best with the TAT-OP (r(2) = 0.98; P = .01), but this relationship weakened substantially after automation (r(2) = 0.29; P > .05), suggesting the laboratory was no longer a factor in ED LOS. The postautomation ED LOS correlated best with ED patient volume (r(2) = 0.88; P = .06). Although laboratories have focused on TAT means for performance assessment, our study suggests TAT-OPs are more clinically relevant benchmarks. Furthermore, our findings suggest that total laboratory automation can effectively improve overall laboratory service reliability and help eliminate the laboratory as a factor in ED LOS. PMID- 16707381 TI - Studies of the extraction of bilirubin from human amniotic fluid. AB - The recovery of unconjugated bilirubin from human amniotic fluid was studied using dichloromethane, chloroform/isopropanol (3:1 vol/vol), and chloroform/ methanol (3:1 vol/vol) extraction of human amniotic fluid that had been supplemented with bilirubin at various concentrations. Results were compared with those obtained with conventional chloroform extraction. Mean recoveries were found to be only 28% for chloroform and 25% for dichloromethane. When the polarity of chloroform was increased by the addition of an alcohol, the mean recovery increased to only 40% for chloroform/isopropanol and 38% for chloroform/ methanol. These results suggest that extraction methods for determination of amniotic fluid "delta OD(450)" (visible spectrophotometric absorbance [optical density] of bilirubin at 450 nm) tend to underestimate the result when compared with the nonextraction (direct-scan) method, on which the Liley Prognostication Chart is based. This finding should be clinically significant, particularly if extraction and direct-scan methods are used to monitor the condition of the same patient. PMID- 16707382 TI - MAL is expressed in a subset of Hodgkin lymphoma and identifies a population of patients with poor prognosis. AB - Classical Hodgkin lymphoma (cHL) and mediastinal (thymic) large B-cell lymphoma (MLBL) have clinical, histopathologic, and molecular genetic similarities. MAL, a gene that encodes a protein associated with lipid rafts in T and epithelial cells, is overexpressed in a majority of MLBLs and has been reported in a minority of cHLs. To study the clinical significance of MAL in cHL, we immunostained 86 cases; 16 cHLs (19%) expressed MAL. Expression correlated with nodular sclerosis subtype, and within this subtype, with grade 2 histology. Univariable analysis revealed association of age of 45 years or older, MAL expression, and an International Prognostic Score of more than 2 with worse failure-free survival. Age of 45 years or older, MAL expression, and stage III or IV were associated with worse overall survival (OS). Cox proportional hazards modeling showed age (P = .04 and P = .03, respectively) and MAL expression (P = .03 and P = .01, respectively) as independent predictors of time to failure-free survival and OS. Stage showed borderline significance in OS (P = .08). MAL expression seems to identify a subset of cHL with an adverse outcome and provides additional evidence for a link between cHL and MLBL. PMID- 16707383 TI - Myeloid sarcoma involving the gynecologic tract: a report of 11 cases and review of the literature. AB - Myeloid sarcoma can involve any anatomic site, but involvement of the gynecologic tract is uncommon. We describe 11 women, 17 to 60 years old, with myeloid sarcoma involving the gynecologic tract, including 5 patients in whom myeloid sarcoma presented as an isolated mass. The uterus was the most frequently involved anatomic site, in 8 patients (5 corpus, 3 cervix). Each neoplasm diffusely infiltrated normal structures, and, cytologically 7 tumors were immature, 3 were differentiated, and 1 was blastic. In 9 cases assessed, immunohistochemical stains showed that all neoplasms were positive for myeloperoxidase and lysozyme; CD117 was positive in 7 of 8 cases, and cytochemical staining for naphthol AS-D chloroacetate was positive in all 6 neoplasms analyzed. Following chemotherapy, complete remission and long-term survival were achieved in a subset of patients, as was particularly true for 2 patients (cases 8 and 10), with complete remission 12.5 and 31 years after diagnosis, respectively. PMID- 16707384 TI - Acute myeloid leukemia with pseudo-Chediak-Higashi anomaly exhibits a specific immunophenotype with CD2 expression. AB - Acute myeloid leukemia (AML) with pseudo-Chediak-Higashi (PCH) anomaly is a rare morphologic entity. We characterized 5 cases by multiparameter flow cytometry and found that in all cases, the blasts aberrantly expressed CD2, a pan-T cell associated marker, in addition to their myeloid-associated markers. In contrast, CD2 was expressed in only 25 (17.9%) of 140 cases of newly diagnosed AML without PCH anomaly. CD2 expression correlated strongly with AML with PCH anomaly (P < .01), suggesting a link between a specific immunophenotypic marker, CD2, and AML with PCH anomaly. PMID- 16707385 TI - Lifting the smokescreen. PMID- 16707386 TI - COPD, asthma and C-reactive protein. PMID- 16707387 TI - The systemic face of airway diseases: the role of C-reactive protein. PMID- 16707388 TI - Much ado about nothing? PMID- 16707389 TI - Sniff nasal inspiratory pressure: simple or too simple? PMID- 16707390 TI - Multiple inflammatory hits and the pathogenesis of severe airway disease. AB - Refractory or difficult-to-control asthma is associated with some clinical and pathological features normally associated with chronic obstructive pulmonary disease (COPD), raising the possibility that there are similarities in their pathogenesis. It is suggested that the coexistence of two or more inflammatory stimuli to the airway (multiple hits) is a key factor leading to the development of more severe airway disease. Airway inflammation in response to chronic inflammatory conditions elsewhere may be a particularly important additional inflammatory stimulus. The "multiple hit" hypothesis for the origins of severe airway disease has important implications for treatment and prevention, since identification and removal of additional inflammatory stimuli may delay progression of the underlying airway disease. PMID- 16707391 TI - High sensitivity C-reactive protein in asthma. AB - Asthma is characterised by chronic inflammation of the airways, but the relevance of high-sensitivity assays for C-reactive protein (hs-CRP), which are known to be a sensitive marker of low-grade systemic inflammation, has not been fully studied in asthma. The objective was to examine serum hs-CRP levels in patients with asthma and their relationship to clinical characteristics and degree of airway inflammation. Serum hs-CRP levels were cross-sectionally examined in steroid naive (n = 22) and steroid-inhaling (n = 23) adult patients with asthma and healthy controls (n = 14). All were nonsmokers. Serum hs-CRP levels were significantly increased in steroid-naive patients (mean+/-sd 1.33+/-1.48 mg.L( 1)) compared with controls (0.21+/-0.30 mg.L(-1)), but not in patients on inhaled corticosteroid. Among steroid-naive patients, serum hs-CRP levels significantly negatively correlated with indices of pulmonary function (forced expiratory volume in one second/forced vital capacity and forced mid-expiratory flow) and positively with sputum eosinophil count. Among patients on inhaled corticosteroid, hs-CRP levels did not correlate with any indices. In conclusion, an increase in serum C-reactive protein levels measured by high-sensitivity assays may be associated with airflow obstruction and airway inflammation, and may serve as a surrogate marker of airway inflammation in asthma. PMID- 16707392 TI - Evaluating the "Leeds criteria" for Pseudomonas aeruginosa infection in a cystic fibrosis centre. AB - Four separate categories of chronic Pseudomonas aeruginosa (Pa) infection in children with cystic fibrosis (CF) have been previously defined, based on airway cultures taken over the previous year. The aim of the present study was to evaluate this definition in the current authors' paediatric and adult CF clinic using clinical, immunological and lung function parameters. During follow-up, out of 193 patients, 55 (34%) CF patients had never been infected with Pa, 27 (17%) were free of Pa, 29 (18%) were intermittently infected and 51 (31%) were chronically infected. Disease severity markers, such as lung function, were significantly worse in the chronic group, especially in the paediatric population. Differences in adult patients were smaller and no longer significant. Pa antibodies differed strongly between the groups, and were very high (mean+/-sd 55.4+/-5.5) and highly statistically significant from all other groups in the chronic group. They were low and different from all other groups in the never group (1.8+/-0.6). Pa antibodies did not differ between the free of Pa and the intermittent group. In conclusion, the current authors confirmed an agreement between Pseudomonas aeruginosa status according to the new definition and clinical status, as well as with the level of Pseudomonas aeruginosa antibodies. PMID- 16707393 TI - Inhibition of mast cell PGD2 release protects against mannitol-induced airway narrowing. AB - Mannitol inhalation increases urinary excretion of 9alpha,11beta-prostaglandin F2 (a metabolite of prostaglandin D2 and marker of mast cell activation) and leukotriene E4. The present study tested the hypothesis that beta2-adrenoreceptor agonists and disodium cromoglycate (SCG) protect against mannitol-induced bronchoconstriction by inhibition of mast cell mediator release. Fourteen asthmatic subjects inhaled mannitol (mean dose 252+/-213 mg) in order to induce a fall in forced expiratory volume in one second (FEV1) of > or = 25%. The same dose was given 15 min after inhalation of formoterol fumarate (24 microg), SCG (40 mg) or placebo. Pre- and post-challenge urine samples were analysed by enzyme immunoassay for 9alpha,11beta-prostaglandin F2 and leukotriene E4. The maximum fall in FEV1 of 32+/-10% on placebo was reduced by 95% following formoterol and 63% following SCG. Following placebo, there was an increase in median urinary 9alpha,11beta-prostaglandin F2 concentration from 61 to 92 ng.mmol creatinine( 1), but no significant increase in 9alpha,11beta-prostaglandin F2 concentration in the presence of either formoterol (69 versus 67 ng.mmol creatinine(-1)) or SCG (66 versus 60 ng.mmol creatinine(-1)). The increase in urinary leukotriene E4 following placebo (from 19 to 31 ng.mmol creatinine(-1)) was unaffected by the drugs. These results support the hypothesis that the drug effect on airway response to mannitol is due to inhibition of mast cell prostaglandin D2 release. PMID- 16707394 TI - Characteristics of tracheostomy phonation valves. AB - Phonation valves are commonly used devices that allow the restoration of speech in tracheostomised patients. However, their use should not compromise the physiological benefit of tracheostomy. Six commercialised phonation valves were studied in a dynamic set-up simulating a respiratory frequency of 20 breaths.min( 1), a tidal volume of 0.5 L and a peak flow rate of 0.5 L.s(-1). Resistance and additional work of breathing (WOB) were calculated. In 10 tracheostomised patients, evaluations using no phonation valve (baseline), and the most and one of the least resistive valves were carried out. Respiratory patterns and gas exchanges were recorded. Inspiratory difficulty was evaluated using the modified Borg scale. Valves displayed a wide array of resistance ranging 1.3-5.9 cmH2O.L( 1).s(-1). Additional WOB varied with a ratio of 4.4 between the best and the worst valve. While the different clinical conditions did not modify respiratory patterns and gas exchanges, a significant effect on the Borg scale rating was observed using ANOVA and post hoc analysis of baseline versus worst valve and one of the best valves versus worst valve. In conclusion, the variety of aerodynamic characteristics of phonation valves should be considered when choosing the device, according to the underlying condition of the patients benefiting from their use. PMID- 16707396 TI - Mandibular advancement and obstructive sleep apnoea: a method for determining effective mandibular protrusion. AB - The objectives of the study were to test the hypotheses that it is possible, during routine polysomnography (PSG), to prospectively identify favourable candidates for mandibular repositioning appliance (MRA) therapy in the treatment of obstructive sleep apnoea (OSA) and to accurately estimate an optimal protrusive distance at which to fabricate the MRA. A series of subjects underwent a remotely controlled mandibular positioner (RCMP) test during PSG monitoring. The ability of the RCMP test to eliminate OSA and the target protrusion at which that occurred was compared with the success of a custom oral MRA in the 33 subjects who completed the protocol. The RCMP test was a success in 15 subjects and a failure in 18 subjects. Appliance therapy was initiated in 38 subjects and completed in 33. MRA therapy was successful at target protrusion in 80% of subjects who had a successful RCMP test and failed in 78% of those who failed the RCMP test. In conclusion the remotely controlled mandibular positioner test outcome demonstrated a statistically significant association with mandibular repositioning appliance outcome. The target protrusion determined during the remotely controlled mandibular positioner test was the effective therapeutic protrusion in subjects with a successful remotely controlled mandibular positioner test. PMID- 16707395 TI - Allopurinol improves endothelial function in sleep apnoea: a randomised controlled study. AB - Increased oxidative stress in obstructive sleep apnoea is thought to contribute to endothelial dysfunction. The objective of this study was to test the hypothesis that inhibition of xanthine oxidase by allopurinol can improve endothelial function in patients with obstructive sleep apnoea. A randomised double-blind placebo-controlled crossover study was performed on 12 patients with moderate-to-severe obstructive sleep apnoea, comparing 300 mg allopurinol daily for 2 weeks with placebo. Endothelial function was assessed using hyperaemia induced flow-mediated vasodilation (FMD) at baseline and following treatment. Plasma malondialdehyde levels were compared in order to assess significant changes in oxidative stress. Baseline FMD correlated significantly with the severity of sleep apnoea and the time spent with an arterial oxygen saturation of <90%. Allopurinol caused a significant increase in FMD compared to placebo (10.4+/-3.2 versus 7.4+/-2.8%, respectively). Plasma malondialdehyde levels were significantly reduced with allopurinol treatment (1.5+/-0.3 versus 1.2+/-0.3 micromol.L(-1)), consistent with reduced oxidative stress. Allopurinol improves endothelial dysfunction in patients with moderate-to-severe obstructive sleep apnoea. These observations suggest that xanthine oxidase contributes significantly to vasodilatory impairment. PMID- 16707397 TI - Effect of prior statin therapy on capillary permeability in the lungs after cardiac or vascular surgery. AB - Cholesterol-lowering statins can ameliorate severe forms of vascular hyperpermeability in experimental studies, and may thereby ameliorate acute lung injury and sepsis. It is unknown whether this also applies to humans. This study aimed to define whether or not prior statin therapy reduces mild post-operative increases in pulmonary capillary protein permeability associated with acute lung injury after cardiac or major vascular surgery. A prospective observational study was performed in an intensive care unit of a university hospital on 64 patients, 37 after elective cardiac and 27 after major vascular surgery, of whom 68 and 44%, respectively, had received prior statin therapy. A mobile probe system was used to measure the pulmonary leak index (PLI), i.e. the transvascular transport rate of gallium-67-radiolabelled transferrin. For all of the patients together, the mean PLI did not differ between the statin and control groups (22.9 versus 24.4 x 10(-3) min(-1)). The prevalence of an elevated PLI was 57% in the statin and 59% in the control group. Subgroup analysis did not reveal significant differences caused by statins in the PLI of these patients. Prior statin therapy neither has an adverse effect on mildly increased pulmonary capillary permeability in patients after cardiac or major vascular surgery nor does it ameliorate this increased capillary permeability. PMID- 16707398 TI - Time course of exhaled hydrogen peroxide and nitric oxide during chemotherapy. AB - This study was designed to assess the effect of differential leukocyte depletion during chemotherapy by monitoring the levels of exhaled hydrogen peroxide H2O2 and nitric oxide (F(eNO)) present. In 39 patients with lung cancer (chronic obstructive pulmonary disorder up to stage II, median forced expiratory volume in one second 78% predicted), measurements were performed before a cycle of therapy (day 1), at least once during the cycle (day 8: n = 34; day 15: n = 19), and afterwards (days 21-29). There were significant changes in the level of H2O2, F(eNO) and peripheral blood cell differentials over the visits. The level of H2O2 was decreased only on day 15, with a median (difference between the upper and lower quartiles) fall of 31 (57)%, while F(eNO) was reduced only on day 8, by 22 (40)%. Neutrophil numbers were unchanged on day 8 and decreased by 59 (48)% on day 15, while monocyte numbers were decreased on day 8 by 87 (39)%. On days 21 29, values had returned to baseline. Taken together with previous findings, the parallel course of levels of exhaled hydrogen peroxide and neutrophil counts suggests that a major part of exhaled hydrogen peroxide is due to neutrophils via the conducting airways. In contrast, the production of exhaled nitric oxide seems to be primarily associated with monocytes. PMID- 16707399 TI - Quantifying physical activity in daily life with questionnaires and motion sensors in COPD. AB - Accurate assessment of the amount and intensity of physical activity in daily life is considered very important due to the close relationship between physical activity level, health, disability and mortality. For this reason, assessment of physical activity in daily life has gained interest in recent years, especially in sedentary populations, such as patients with chronic obstructive pulmonary disease (COPD). The present article aims to compare and discuss the two kinds of instruments more commonly used to quantify the amount of physical activity performed by COPD patients in daily life: subjective methods (questionnaires, diaries) and motion sensors (electronic or mechanical methods). Their characteristics are summarised and evidence of their validity, reliability and sensitivity is discussed, when available. Subjective methods have practical value mainly in providing the patients' view on their performance in activities of daily living and functional status. However, care must be taken when using subjective methods to accurately quantify the amount of daily physical activity performed. More accurate information is likely to be available with motion sensors rather than questionnaires. The selection of which motion sensor to use for quantification of physical activity in daily life should depend mainly on the purpose of its use. PMID- 16707400 TI - Lymphangioleiomyomatosis. AB - Lymphangioleiomyomatosis (LAM) is a rare disease of the lungs and lymphatics, which can occur sporadically or in association with tuberous sclerosis. LAM almost exclusively affects females, generally developing before the menopause. The disease is characterised by progressive pulmonary cystic change, recurrent pneumothorax, chylous pleural collections and, in most cases, progressive respiratory failure. Abdominal manifestations include lymphadenopathy, cystic lymphatic masses (lymphangioleiomyomas), chylous ascites and angiomyolipoma (a benign tumour). Survival in LAM is approximately 70% at 10 yrs, although this is highly variable since long-term survivors have been described. Diagnosis is made by a combination of clinical features and computed tomography scanning or, in cases of doubt, lung biopsy. In patients with rapidly progressive disease, hormone treatment (predominantly progesterone) has been used, although no firm evidence supports its use. Otherwise, treatment is aimed at complications including pneumothorax, chylous collections and extrapulmonary manifestations. The only treatment for severe LAM is currently lung transplantation. Recent developments in the cell biology of lymphangioleiomyomatosis have shown that these patients have somatic mutations in the genes linked to tuberous sclerosis and that rapamycin may correct the resulting cellular abnormality. Trials of rapamycin in lymphangioleiomyomatosis are currently underway and offer hope of evidence-based treatment for the disease. PMID- 16707401 TI - Hepatic veno-occlusive disease due to tacrolimus in a single-lung transplant patient. AB - Hepatic veno-occlusive disease is defined as nonthrombotic fibrous obliterative endophlebitis of small centrilobular hepatic venules. Clinically, patients present with elevated liver enzymes and a triad of jaundice, hepatomegaly and ascites. Although reported as a complication of other solid organ and stem cell transplantation, there have been no reported cases to date of veno-occlusive disease following lung transplantation. The present authors report a case of veno occlusive disease following single-lung transplantation in a patient on a triple drug immunosuppressive regimen composed of tacrolimus, mycophenolate mofetil and prednisone. The diagnosis was established by transjugular liver biopsy and by discontinuing tacrolimus; there was clinical regression of symptoms and serological return to baseline. PMID- 16707402 TI - Obstructive ventilatory defect with normal forced expiratory volume in one second/vital capacity ratio. PMID- 16707403 TI - Inhaler technique blind spot. PMID- 16707404 TI - External validity of randomised controlled trials in idiopathic pulmonary fibrosis. PMID- 16707405 TI - Training and ventilatory assistance in chronic obstructive pulmonary disease. PMID- 16707406 TI - Man versus machine: is there an optimal method for QT measurements in thorough QT studies? AB - Electrocardiographic (ECG) recordings from 3 placebo-controlled thorough QT healthy volunteer studies were used to compare QT intervals obtained by manual measurement with those generated by ECG machines. The effect of the positive control was compared to placebo at each time point for data obtained from both sources. Both manual and automated techniques consistently demonstrated statistically significant prolongation of QTcF with the positive controls. The proportion of outlier values was small for both methods. The pairwise comparison between manual and automated uncorrected QT intervals demonstrated clear differences, with intervals derived from one machine on average 16 to 19 milliseconds shorter and from the other 7 milliseconds longer than the manually measured QT intervals, but these differences disappeared when analyzing QT change from baseline. Both manual and automated, commercially available QT algorithms demonstrated small statistically significant effects on the QTc interval induced by positive controls. PMID- 16707407 TI - National Veterans Health Administration hospitalizations for syncope compared to acute myocardial infarction, fracture, or pneumonia in community-dwelling elders: outpatient medication and comorbidity profiles. AB - The authors used 2 national Veterans Health Administration databases to identify outpatient medications and all 30 Elixhauser comorbidities for 2579 unique patients, age 65+ years, hospitalized for syncope in fiscal year 2004. For comparison, we identified other elderly patients hospitalized with acute myocardial infarction (N = 4491), fracture (N = 2797), or pneumonia (N = 9473). The categories of medications included drugs that affect the cardiovascular, central nervous, or the muscular skeletal system. The most notable differences between syncope compared to acute myocardial infarction patients occurred in central nervous system drugs in anticonvulsants/barbiturates, antidepressants, antihistamine/antinauseants, antipsychotics, and cholinesterase inhibitors (P < .0018). Comparing syncope patients with fracture patients, the central nervous medication profile was similar, but the cardiovascular medication profile differed (P < .0018); their hypertension comorbidities also differed (60.45% vs 46.34%); (P < .0016). These findings indicate significant potential associations that warrant further study. Studies linking national outpatient medications to hospitalizations for specific conditions can foster the development of more proactive pharmacovigilance systems. PMID- 16707408 TI - Pharmacokinetics and safety of montelukast in children aged 3 to 6 months. AB - The single-dose population estimate of the area under the concentration-time curve (AUC(pop)) from time zero to infinity (AUC(0-infinity)), maximum plasma concentration (C(max)), and time to C(max) (t(max)) of montelukast 4-mg oral granules were investigated in infants aged 3 to 6 months. Montelukast concentrations were quantitated after a single 4-mg dose of montelukast oral granules. Pharmacokinetic parameters were determined using a population-based approach with a nonlinear mixed-effect, 1-compartment model with first-order absorption and elimination. Ninety-five percent confidence intervals for the AUC(pop) ratio (3 to 6 months/6 to 24 months) were determined. Safety and tolerability were assessed. Montelukast 4-mg oral granules in children 3 to 6 months of age yielded systemic exposure (AUC(pop) = 3644.3 +/- 481.5 ng x h/mL) similar to that observed in children aged 6 to 24 months (3226.6 +/- 250.0 ng x h/mL). Systemic exposure after a 4-mg dose of montelukast as oral granules is similar in children aged 3 to 6 months and 6 to 24 months. PMID- 16707409 TI - The effects of ergoloid mesylates and ginkgo biloba on the pharmacokinetics of ticlopidine. AB - Ticlopidine is sometimes coadministered with ergoloid mesylates or ginkgo biloba in clinical situations. Our objective was to examine the effect of ergoloid mesylates and ginkgo biloba on ticlopidine pharmacokinetics. Ticlopidine, ergoloid mesylates, and ginkgo biloba significantly inhibited the organic anion transporting polypeptide (OATP-B)-mediated uptake of [(3)H]-estrone-3-sulfate in a concentration-dependent manner. When ergoloid mesylates was coadministered with ticlopidine, the ticlopidine area under the plasma drug concentration-time profile (AUC) from 0 to 12 hours was decreased 30% and the peak plasma drug concentration (C(max)) was decreased 29%, compared with ticlopidine administration alone. There were no significant changes in the pharmacokinetic parameters of ticlopidine when it was coadministered with ginkgo biloba. In summary, ergoloid mesylates is a more potent inhibitor of OATP-B than is ginkgo biloba, and it can reduce the oral bioavailability of drugs transported by OATP B. Ergoloid mesylates markedly decreased the AUC and C(max) of ticlopidine, probably by inhibiting the OATP-B-mediated uptake of ticlopidine during the intestinal absorption phase. The results support a new model of intestinal drug drug interaction. PMID- 16707410 TI - An open-label, parallel group study investigating the effects of age and gender on the pharmacokinetics of the once-daily glucagon-like peptide-1 analogue liraglutide. AB - Liraglutide is a once-daily glucagon-like peptide-1 analogue being developed for the treatment of type 2 diabetes. The aim of this study was to investigate the effect of age and gender on the pharmacokinetics of liraglutide. Eight male and 8 female subjects were recruited from an 18- to 45-year-old group and an over-65 year-old group, respectively. All subjects received a single subcutaneous dose of 1.0 mg liraglutide. The area under the liraglutide plasma concentration curve from time 0 to last quantifiable concentration adjusted for body weight (significant covariate; P = .001) was found to be equivalent in young and elderly subjects (primary end point), with an estimated ratio of 0.94 (90% confidence interval, 0.84-1.06; P = .39). No significant impact of gender was observed (P = .38; estimated ratio, 1.08; 90% confidence interval, 0.93-1.26). Adverse events were of mild or moderate severity. The most frequently reported events were headache, vomiting, and nausea. When adjusted for body weight, no effect of gender or age was found on the pharmacokinetics of liraglutide. PMID- 16707411 TI - Pharmacokinetics, tolerability, and performance of a novel matrix transdermal delivery system of fentanyl relative to the commercially available reservoir formulation in healthy subjects. AB - A novel transdermal formulation of fentanyl-containing dipropylene glycol droplets dispersed in a silicone matrix with a rate-controlling membrane was developed. Healthy male subjects (n = 24) received repeated 72-hour applications of fentanyl (50 mug/h) as the novel matrix and the conventional reservoir formulations in a randomized, 2-way crossover study. Blood samples were collected, and serum concentrations of fentanyl were assayed using liquid chromatography with mass spectrometry detection. The mean area under the curve (AUCtau) and peak concentrations (C(max)) of the matrix formulation were 84 838 pg.h/mL and 1680 pg/mL, respectively. Ratio and 90% confidence intervals of AUCtau and C(max) between the 2 formulations were within 80% to 125%. Adherence of the matrix formulation was higher than the reservoir formulation (62.5 vs 56.2%, P < .0001), without affecting skin irritation. Vital signs and adverse events of the 2 formulations were similar in nature and frequency. The novel matrix formulation displayed enhanced adherence and resulted in similar pharmacokinetics and tolerability as the reservoir formulation. PMID- 16707412 TI - Bioequivalence of liquid and reconstituted lyophilized etanercept subcutaneous injections. AB - The objective of this study was to compare the pharmacokinetics of liquid and reconstituted lyophilized etanercept. This single-center, open-label study had a 2-period crossover design in which 36 healthy subjects were randomly assigned in a 1:1 ratio to etanercept (liquid/lyo or lyo/liquid). The treatments were separated by 28 days. Blood samples were obtained predose and at 10 predetermined time points postdose. Serum concentrations were determined by enzyme-linked immunosorbent assay. Noncompartmental pharmacokinetic parameters were analyzed using a standard crossover analysis of variance model. Thirty-three subjects completed both treatment periods. Geometric mean values (adjusted) of area under the serum drug concentration-time curve from time zero to the time of the final quantifiable sample, area under the serum drug concentration-time curve from time zero to infinity, and maximum concentration obtained with the 50-mg/mL liquid etanercept injection were 93.0%, 90.7%, and 98.5% of the respective parameters for 2 injections of 25 mg/mL reconstituted formulation. All associated confidence intervals were within the predefined equivalence interval of 80% to 125%. No differences in safety profiles of the 2 formulations were apparent. Liquid etanercept was bioequivalent to the reconstituted lyophilized etanercept formulation. PMID- 16707413 TI - Development of a semimechanistic model to describe the pharmacokinetics of gentamicin in patients receiving hemodialysis. AB - The aim of this study was to ascertain the most suitable dosing schedule for gentamicin in patients receiving hemodialysis. We developed a model to describe the concentration-time course of gentamicin in patients receiving hemodialysis. Using the model, an optimal dosing schedule was evaluated. Various dosing regimens were compared in their ability to achieve maximum concentration (C(max), > or = 8 mg/L) and area under the concentration time-curve (AUC > or = 70 mg x h/L and < or = 120 mg x h/L per 24 hours). The model was evaluated by comparing model predictions against real data collected retrospectively. Simulations from the model confirmed the benefits of predialysis dosing. The mean optimal dose was 230 mg administered immediately before dialysis. The model was found to have good predictive performance when simulated data were compared to data observed in real patients. In summary, a model was developed that describes gentamicin pharmacokinetics in patients receiving hemodialysis. Predialysis dosing provided a superior pharmacokinetic profile than did postdialysis dosing. PMID- 16707414 TI - Comparison of maximum drug concentration and area under the time-concentration curve between humans and animals for oral and intravenous investigational drugs. AB - The study compared maximum drug concentration (C(max)) and area under the time concentration curve (AUC) after normalization of doses to body weight and to body surface area and developed relationships for C(max) and AUC between humans and animals for 75 oral and 10 intravenous investigational drugs. For the oral drugs, animal-human ratios of C(max) were different among animals in both normalizations. Surface area-normalized AUC ratios were not different, whereas weight-normalized ones were different. For both normalizations for intravenous drugs, AUC ratios were not different. Drugs exhibiting 1/10 or smaller ratios tended to have low bioavailability. Regression of the relationships for dose normalized C(max) and AUC transformed logarithmically between humans and animals were significant for the drugs with relatively high bioavailability. As approaches for predicting human C(max) and AUC from animals, surface area normalization seems to surpass weight normalization, and the equation obtained can be applied to drugs with high bioavailability. PMID- 16707415 TI - Clinical pharmacology of lasofoxifene in Japanese and white postmenopausal women. PMID- 16707416 TI - Role of macrophage inhibitory cytokine-1 in tumorigenesis and diagnosis of cancer. AB - Macrophage inhibitory cytokine-1 (MIC-1), a transforming growth factor-beta superfamily cytokine, is involved in tumor pathogenesis, and its measurement can be used as a clinical tool for the diagnosis and management of a wide range of cancers. Although generally considered to be part of the cell's antitumorigenic repertoire, MIC-1 secretion, processing, and latent storage suggest a complex, dynamic variability in MIC-1 bioavailability in the tumor microenvironment, potentially modulating tumor progression and invasiveness. PMID- 16707417 TI - Regulatory T cells and toll-like receptors in cancer therapy. AB - T regulatory (Treg) cells that suppress immune responses may limit the efficiency of cancer immunotherapy. Recent findings indicate that Toll-like receptors (TLR) directly regulate the suppressive activity of Treg cells. Linking TLR signaling to the functional control of Treg cells may offer new opportunities to improve the outcome of cancer immunotherapy by coadminstration of certain TLR ligands. PMID- 16707418 TI - Bcl-w promotes gastric cancer cell invasion by inducing matrix metalloproteinase 2 expression via phosphoinositide 3-kinase, Akt, and Sp1. AB - Given a previous report that Bcl-w is expressed in gastric cancer cells, particularly in those of an infiltrative morphology, we investigated whether Bcl w expression influences the invasiveness of gastric cancer cells. To accomplish this, Bcl-w was overexpressed in adherent types of gastric adenocarcinoma cell lines, and this was found to result in an increase in their migratory and invasive potentials. These effects were not induced when Bcl-2 was overexpressed in the same cell types. Consistently, Bcl-w, but not Bcl-2, overexpression increased matrix metalloproteinase-2 (MMP-2) expression, and synthetic or natural inhibitors of MMP-2 abolished Bcl-w-induced cell invasion. Bcl-w overexpression also activated phosphoinositide 3-kinase (PI3K), Akt, and Sp1, and the blocking effects of each of these components using pharmacologic inhibitors, dominant negative mutants, or small interfering RNA abolished the ability of Bcl-w to induce MMP-2 and cell invasion. The inhibition of PI3K/Akt signaling also prevented Sp1 activation. Overall, our data suggest that Bcl-w, which was previously shown to enhance gastric cancer cell survivability, also promotes their invasiveness by inducing MMP-2 expression via the sequential actions of PI3K, Akt, and Sp1. PMID- 16707419 TI - Targeting Aurora kinases for the treatment of prostate cancer. AB - Inappropriate expression of the Aurora kinases can induce aberrant mitosis, centrosome irregularities, and chromosomal instability, which lead to anueploidy and cell transformation. Here, we report that Aurora-A and Aurora-B are highly expressed in primary human and mouse prostate cancers and prostate cancer cell lines. In clinical samples, levels of Aurora-A and Aurora-B were significantly elevated in prostatic intraepithelial neoplasia lesions and prostate tumors when compared with the non-neoplastic samples. Interestingly, expression of Aurora-A in non-neoplastic prostates correlated with seminal vesicle invasion (rho = 0.275, P = 0.0169) and in prostate tumor with positive surgical margins (rho = 0.265, P = 0.0161). In addition, nuclear expression of Aurora-B in prostatic intraepithelial neoplasia lesions correlated with clinical staging of the tumor (rho = -0.4, P = 0.0474) whereas cytoplasmic expression in tumors correlated with seminal vesicle invasion (rho = 0.282, P = 0.0098). Cell lines and primary tumors derived from the TRAMP model were also found to express high levels of Aurora-A and Aurora-B. When human PC3, LNCaP, and mouse C1A cells were treated with the potent Aurora kinase inhibitor VX680, which attenuates phosphorylation of histone H3, cancer cell survival was reduced. VX680 could further reduce cell viability >2-fold when used in combination with the chemotherapy drug doxorubicin. Our findings support a functional relationship between Aurora kinase expression and prostate cancer and the application of small-molecule inhibitors in therapeutic modalities. PMID- 16707420 TI - Evidence of linkage to chromosome 9q22.33 in colorectal cancer kindreds from the United Kingdom. AB - About 30% of all colorectal cancers are thought to have a genetic basis and the known predisposing genes can only account for a small fraction of cases. A previous report suggested that a colorectal cancer candidate gene, explaining at least 20% of colorectal cancer cases with family history, was located within a 25 cM region on chromosome 9q22.2-q31.3. We typed 16 polymorphic markers encompassing the region of putative linkage in 57 colorectal tumor families from the United Kingdom. Known Mendelian syndromes had been excluded. We found suggestive evidence of linkage, as positive parametric (HLOD = 1.23) and nonparametric (NPL = 1.21, P = 0.11) LOD scores were obtained by analysis of the whole family set. Enrichment for cases with a priori genetic etiology by analyzing families with at least one person affected at <45 years of age (n = 39 families) gave a maximum multipoint NPL score of 2.65 (P = 0.007). In this group, significant NPL scores >1.67 (P < 0.05) were found in a 6.5 cM region between D9S1851 and D9S277. With a more stringent threshold (NPL>2.4, P < 0.01), the linked region was 1.7 cM between D9S971 and D9S272/D9S173. Exclusion from the analysis of kindreds with a phenotype of multiple polyposis also found evidence of linkage in the same region (NPL = 2.47 at close to D9S277, P = 0.009). The type I transforming growth factor-beta receptor, a prime candidate gene, was excluded as a cause of disease. The results presented here further support the existence of a colorectal cancer susceptibility gene on chromosome 9q and refine its likely location. PMID- 16707421 TI - Novel 5' untranslated region variants of BCRP mRNA are differentially expressed in drug-selected cancer cells and in normal human tissues: implications for drug resistance, tissue-specific expression, and alternative promoter usage. AB - To investigate transcriptional activation of the breast cancer resistance protein gene (BCRP/ABCG2), we examined the 5' untranslated region of BCRP mRNA in cell lines with high BCRP transcriptional activity and in normal tissues. Human choriocarcinoma cells with high endogenous BCRP expression (JAR and BeWo) and human cancer cells (MCF-7 and Igrov1) and their BCRP-overexpressing, drug selected, multidrug-resistant derivatives (MCF-7/AdrVp, Igrov1/MX3, and Igrov1/T8) were studied. Rapid amplification of 5'-cDNA ends-PCR (5'RACE-PCR) revealed at least three novel forms of the untranslated exon 1 (E1a, E1b, and E1c) that are spliced to a common exon 2, with differential expression of these splice variants in the drug-selected cell lines. Additionally, sequence analysis of the 5'RACE-PCR products revealed multiple transcriptional start sites for each variant, particularly in the drug-selected cells. The E1c isoform predominated in drug-selected MCF-7 cell lines and was translated more efficiently in MCF-7 cells than the E1a isoform. Varying patterns of expression of the exon 1 isoforms were observed in a variety of human tissues, suggesting that tissue-specific alternative promoters of BCRP exist. In summary, we find that BCRP overexpression in the drug-selected cells is accompanied by multiple transcriptional start sites and predominance of the more efficiently translated E1c isoform. The exon 1 variation we observe suggests that alternative promoters of the BCRP gene exist. PMID- 16707422 TI - Evolution of the androgen receptor pathway during progression of prostate cancer. AB - The present work focused on the potential involvement of selective adaptations of the androgen receptor pathway in the initiation and progression of prostate cancer. We defined the androgen receptor pathway by selecting 200 genes that were androgen responsive in prostate cancer cell lines and/or xenografts. This androgen receptor pathway gene signature was then used for profiling prostate cancer xenografts and patient-derived samples. Approximately half of the androgen receptor pathway genes were up-regulated in well-differentiated prostate cancer compared with normal prostate. Functionally distinct parts of the androgen receptor pathway were specifically down-regulated in high-grade cancers. Unexpectedly, metastases have down-regulated the vast majority of androgen receptor pathway genes. The significance of this progressive down-regulation of androgen receptor pathway genes was shown for a few androgen receptor-regulated genes. Lower mRNA expression of HERPUD1, STK39, DHCR24, and SOCS2 in primary prostate tumors was correlated with a higher incidence of metastases after radical prostatectomy. HERPUD1 mRNA expression predicted the occurrence of metastases almost perfectly. In vitro experiments showed that overexpression of the stress response gene HERPUD1 rapidly induces apoptosis. Based on the functions of the genes within the distinct subsets, we propose the following model. Enhanced androgen receptor activity is involved in the early stages of prostate cancer. In well-differentiated prostate cancer, the androgen receptor activates growth-promoting as well as growth-inhibiting and cell differentiation genes resulting in a low growth rate. The progression from low-grade to high grade prostate carcinoma and metastases is mediated by a selective down regulation of the androgen receptor target genes that inhibit proliferation, induce differentiation, or mediate apoptosis. PMID- 16707423 TI - Identification of novel target genes by an epigenetic reactivation screen of renal cancer. AB - Aberrant promoter hypermethylation is a common mechanism for inactivation of tumor suppressor genes in cancer cells. To generate a global profile of genes silenced by hypermethylation in renal cell cancer (RCC), we did an expression microarray-based analysis of genes reactivated in the 786-0, ACHN, HRC51, and HRC59 RCC lines after treatment with the demethylating drug 5-aza-2 deoxycytidine and histone deacetylation inhibiting drug trichostatin A. Between 111 to 170 genes were found to have at least 3-fold up-regulation of expression after treatment in each cell line. To establish the specificity of the screen for identification of genes, epigenetically silenced in cancer cells, we validated a subset of 12 up-regulated genes. Three genes (IGFBP1, IGFBP3, and COL1A1) showed promoter methylation in tumor DNA but were unmethylated in normal cell DNA. One gene (GDF15) was methylated in normal cells but more densely methylated in tumor cells. One gene (PLAU) showed cancer cell-specific methylation that did not correlate well with expression status. The remaining seven genes had unmethylated promoters, although at least one of these genes (TGM2) may be regulated by RASSF1A, which was methylated in the RCC lines. Thus, we were able to show that up-regulation of at least 6 of the 12 genes examined was due to epigenetic reactivation. The IGFBP1, IGFBP3, and COL1A1 gene promoter regions were found to be frequently methylated in primary renal cell tumors, and further study will provide insight into the biology of the disease and facilitate translational studies in renal cancer. PMID- 16707424 TI - Identification of candidate alkylator-induced cancer susceptibility genes by whole genome scanning in mice. AB - Secondary malignancies are a serious adverse consequence of alkylator chemotherapy. The risk of developing an alkylator-associated malignancy is influenced by genetic background, although the relevant genetic factors are poorly understood. To screen for novel susceptibility factors, we established a mouse model of alkylator-induced malignancy. We exposed mice from 20 inbred strains to the prototypical alkylating agent, N-nitroso-N-ethylurea (ENU). ENU was a potent carcinogen in many of the strains tested, inducing 140 tumors in 240 ENU-treated mice (66% incidence of at least one tumor in evaluable mice), compared with a background incidence of 8% spontaneous tumors in 240 strain-, age , and sex-matched control mice (relative risk, 8.4; P < 0.0001). A wide variety of tumor histologies were noted, including epithelial carcinomas, soft tissue sarcomas, and hematopoietic tumors. Cancer susceptibility was a heritable trait for the most common tumor types, lung adenocarcinoma (H(2) = 0.25), T cell lymphoma (H(2) = 0.19), and myeloid malignancies (H(2) = 0.10). Quantitative trait locus mapping identified regions on chromosomes 3, 6, 9, and 15 containing candidate genes associated with lung adenoma, lung carcinoma, and lymphoma susceptibility. This novel mouse model recapitulates many features of human alkylator-associated cancer and supports the hypothesis that susceptibility to this syndrome is influenced by inherited polymorphisms that could be used to make informed clinical treatment decisions. PMID- 16707425 TI - BRCC36 is essential for ionizing radiation-induced BRCA1 phosphorylation and nuclear foci formation. AB - We have previously reported the identification and characterization of a novel BRCA1/2 interacting protein complex, BRCC (BRCA1/2-containing complex). BRCC36, one of the proteins in BRCC, directly interacts with BRCA1, and regulates the ubiquitin E3 ligase activity of BRCC. Importantly, BRCC36 is aberrantly expressed in the vast majority of breast tumors, indicating a potential role in the pathogenesis of this disease. To further elucidate the functional consequence of abnormal BRCC36 expression in breast cancer, we have done in vivo silencing studies using small interfering RNAs targeting BRCC36 in breast cancer cell lines, i.e., MCF-7, ZR-75-1, and T47D. Knock-down of BRCC36 alone does not affect cell growth, but when combined with ionizing radiation (IR) exposure, it leads to an increase in the percentage of cells undergoing apoptosis when compared with the small interfering RNA control group in breast cancer cells. Immunoblot analysis shows that inhibition of BRCC36 has no effect on the activation of ATM, expression of p21 and p53, or BRCA1-BARD1 interaction following IR exposure. Importantly, BRCC36 depletion disrupts IR-induced phosphorylation of BRCA1. Immunofluorescent staining of BRCA1 and gamma-H2AX indicates that BRCC36 depletion prevents the formation of BRCA1 nuclear foci in response to DNA damage in breast cancer cells. These results show that down-regulation of BRCC36 expression impairs the DNA repair pathway activated in response to IR by inhibiting BRCA1 activation, thereby sensitizing breast cancer cells to IR induced apoptosis. PMID- 16707426 TI - Antiangiogenic antithrombin induces global changes in the gene expression profile of endothelial cells. AB - Antithrombin, a serpin family protease inhibitor crucial to hemostasis, acquires antiangiogenic properties on undergoing conformational alterations induced by limited proteolysis or elevated temperature. To better understand the biochemical mechanisms underlying antithrombin antiangiogenic activity, we did genome-wide expression profiling, coupled with quantitative reverse transcription-PCR, Northern blot, and Western blot analyses, to characterize the gene expression patterns that are induced by antiangiogenic antithrombin in cultured primary human umbilical vein endothelial cells. Overall, 35 genes with significantly increased expression and 93 genes with significantly reduced expression (> or =2 fold changes) due to antiangiogenic antithrombin treatment were identified. More than half of the down-regulated genes have well-established proangiogenic functions in endothelial cells, including cell-surface and matrix proteoglycans (e.g., perlecan, biglycan, and syndecans 1 and 3) and mitogenesis-related signaling proteins (e.g., mitogen-activated protein kinase 3, signal transducers and activators of transcription 2, 3, and 6, and early growth response factor 1). In contrast, most up-regulated genes (e.g., caspase-3, p21, tissue inhibitor of metalloproteinases 1, 2, and 3, and adenomatosis polyposis coli) are known for their antiangiogenic functions which include the promotion of cell apoptosis and cell cycle arrest and the inhibition of tumor growth and metastasis. These results show that the antiangiogenic activity of antithrombin is mediated at least in part by a global genetic reprogramming of endothelial cells and strongly implicate an endothelial cell ligand-receptor signaling mechanism in this reprogramming. PMID- 16707427 TI - Identification of a novel germ line variant hotspot mutant p53-R175L in pediatric adrenal cortical carcinoma. AB - Hotspot mutations in the p53 tumor suppressor gene result in the disruption of DNA contact points or alter the overall structure of the protein to prevent DNA binding. When inherited, hotspot mutants are associated with Li-Fraumeni syndrome (LFS), a familial cancer predisposition. One of the most common hotspot mutations occurs at codon 175, resulting in an arginine to histidine substitution. We have identified a novel germ line variant of the 175 mutant (Arg to Leu; R175L) in a pediatric patient who developed adrenal cortical carcinoma. Surprisingly, the family is not tumor prone or associated with LFS. In vitro, the R175L mutant displayed an attenuated tumor suppressor activity in the regulation of transcription, colony formation, and apoptosis when compared with wild-type p53 and the R175H mutant. These findings suggest that p53-R175L retains sufficient activity to suppress LFS, but not adrenal cortical carcinoma. Therefore, not all hotspot mutants are functionally equivalent and the biochemical nature of the mutant may significantly influence clinical outcome. The implications of these results for genetic counseling are discussed. PMID- 16707428 TI - Genetic predisposition to fiber carcinogenesis causes a mesothelioma epidemic in Turkey. AB - Malignant mesothelioma in the western world is often associated with asbestos exposure. It is a relatively rare cancer that causes approximately 2,500 deaths yearly in the United States and 1,000 deaths yearly in the United Kingdom. In contrast, among people born in the Cappadocian (Turkey) villages of Tuzkoy, Karain, and "Old" Sarihidir, approximately 50% of deaths are caused by malignant mesothelioma. This epidemic has been attributed to erionite exposure, a type of fibrous zeolite mineral commonly found in this area of Turkey. In these three villages, malignant mesothelioma occurs in certain houses but not in others. The hypothesis was that a unique and more carcinogenic erionite was present in certain houses and caused malignant mesothelioma. We determined the X-ray diffraction pattern and the crystal structure of erionite from malignant mesothelioma villages and compared the results with the erionite samples from nearby non-malignant mesothelioma villages and from the United States. We found the same type of erionite in Cappadocian villages, with or without a malignant mesothelioma epidemic, in households with high or no incidence of malignant mesothelioma and in the United States. Pedigree studies of the three malignant mesothelioma villages showed that malignant mesothelioma was prevalent in certain families but not in others. When high-risk malignant mesothelioma family members married into families with no history of it, malignant mesothelioma appeared in the descendants. Genetically predisposed family members born and raised outside the malignant mesothelioma villages did not seem to develop malignant mesothelioma. In summary, pedigree and mineralogical studies indicate that the malignant mesothelioma epidemic is caused by erionite exposure in genetically predisposed individuals. This is the first time that genetics is shown to influence mineral fiber carcinogenesis. PMID- 16707429 TI - Role for Brm in cell growth control. AB - Recently, we have shown implication of Brm, the catalytic subunit of the SWI/SNF chromatin remodeling complex, in repression of cyclin A expression in quiescent cells. Here, we have examined the fate of cells lacking Brm throughout the cycle. We find that despite elevated levels of cyclins A and E, these cells can respond to serum starvation, however, without reaching a canonical G(0) phase as they continue to express high levels of c-Myc and have an abnormally large average size. The response to serum starvation can be correlated with increased levels of Rb proteins p130 and p107 as well as increased association of p27 with the cyclin dependent kinases, possibly compensating for the higher levels of G(1) cyclins by reducing their associated kinase activity. After serum stimulation, reentry into the cycle occurs normally, but the S phase is delayed and shorter. In addition, the M phase has an increased duration, and we observed frequent faulty chromosome segregation events in anaphase. Altogether, our data suggest that cells can partially overcome the absence of Brm by activating several compensatory mechanisms to control the cell cycle. However, they remain profoundly affected, unable to enter a canonical quiescent state, presenting a shorter S phase, and finally unable to perform correct chromosome segregation. PMID- 16707430 TI - Silencing of bidirectional promoters by DNA methylation in tumorigenesis. AB - CpG island methylation within promoters is known to silence individual genes in cancer. The involvement of this process in silencing gene pairs controlled by bidirectional promoters is unclear. In a screen for hypermethylated CpG islands in cancer, bidirectional promoters constituted 25.2% of all identified promoters, which matches with the genomic representation of bidirectional promoters. From the screen, we selected three bidirectional gene pairs for detailed analysis, WNT9A/CD558500, CTDSPL/BC040563, and KCNK15/BF195580. Levels of mRNA of all three pairs of genes were inversely correlated with the degree of promoter methylation in multiple cancer cell lines. Hypomethylation of these promoters induced by 5 aza-2'-deoxycytidine treatment reactivated or enhanced gene expression bidirectionally. The bidirectional nature of the WNT9A/CD558500 promoter was confirmed by luciferase assays, and hypermethylation down-regulated expression of both genes in the pair. Methylation of WNT9A/CD558500 and CTDSPL/BC040563 promoters occurs frequently in primary colon cancers and acute lymphoid leukemias (ALL), respectively, and methylation was correlated with decreased gene expression in ALL patient samples. Our study shows that hypermethylation of bidirectional promoter-associated CpG island silences two genes simultaneously, a property that should be taken into account when studying the functional consequences of hypermethylation in cancer. PMID- 16707431 TI - Activator protein-2 overexpression accounts for increased insulin receptor expression in human breast cancer. AB - Various studies have shown that the insulin receptor (IR) is increased in most human breast cancers, and both ligand-dependent malignant transformation and increased cell growth occur in cultured breast cells overexpressing the IR. However, although numerous in vivo and in vitro observations have indicated an important contributory role for the IR in breast cancer cell biology, the molecular mechanisms accounting for increased IR expression in breast tumors have not previously been elucidated. Herein, we did immunoblot analyses of nuclear protein from cultured breast cancer cells and normal and tumoral tissues from breast cancer patients combined with promoter studies by using a series of human wild-type and mutant IR promoter constructs. We provide evidence that IR overexpression in breast cancer is dependent on the assembly of a transcriptionally active multiprotein-DNA complex, which includes the high mobility group A1 (HMGA1) protein, the developmentally regulated activator protein-2 (AP-2) transcription factor and the ubiquitously expressed transcription factor Sp1. In cultured breast cancer cells and human breast cancer specimens, the expression of AP-2 was significantly higher than that observed in cells and tissues derived from normal breast, and this overexpression paralleled the increase in IR expression. However, AP-2 DNA-binding activity was undetectable with the IR gene promoter, suggesting that transactivation of this gene by AP-2 might occur indirectly through physical and functional cooperation with HMGA1 and Sp1. Our findings support this hypothesis and suggest that in affected individuals, hyperactivation of the AP-2 gene through the overexpression of IR may play a key role in breast carcinogenesis. PMID- 16707432 TI - Differentially active origins of DNA replication in tumor versus normal cells. AB - Previously, a degenerate 36 bp human consensus sequence was identified as a determinant of autonomous replication in eukaryotic cells. Random mutagenesis analyses further identified an internal 20 bp of the 36 bp consensus sequence as sufficient for acting as a core origin element. Here, we have located six versions of the 20 bp consensus sequence (20mer) on human chromosome 19q13 over a region spanning approximately 211 kb and tested them for ectopic and in situ replication activity by transient episomal replication assays and nascent DNA strand abundance analyses, respectively. The six versions of the 20mer alone were capable of supporting autonomous replication of their respective plasmids, unlike random genomic sequence of the same length. Furthermore, comparative analyses of the endogenous replication activity of these 20mers at their respective chromosomal sites, in five tumor/transformed and two normal cell lines, done by in situ chromosomal DNA replication assays, involving preparation of nascent DNA by the lambda exonuclease method and quantification by real-time PCR, showed that these sites coincided with chromosomal origins of DNA replication in all cell lines. Moreover, a 2- to 3-fold higher origin activity in the tumor/transformed cells by comparison to the normal cells was observed, suggesting a higher activation of these origins in tumor/transformed cell lines. PMID- 16707433 TI - MDM2 SNP309 accelerates tumor formation in a gender-specific and hormone dependent manner. AB - The importance of the p53 stress response pathway in the suppression of tumor formation is well documented. In a previous report, a single nucleotide polymorphism (SNP309 T/G) was found in the promoter of the MDM2 gene resulting in higher levels of MDM2 RNA and protein and, consequently, in the attenuation of the p53 pathway both in vitro and in vivo. As the SNP309 locus is found in a region of the MDM2 promoter, which is regulated by hormonal signaling pathways, and the G-allele of SNP309 increases the affinity of a well-described cotranscriptional activator of nuclear hormone receptors (i.e., Sp1), the hypothesis that the SNP309 locus could alter the effects of hormones on tumorigenesis was tested in vivo in humans. Data obtained from patients with three different sporadic cancers, from four independent case studies, support this hypothesis, providing an example for the genetic basis of gender differences in cancer and showing that the genotype at a specific locus can affect how hormones, like estrogen, affect tumorigenesis in humans. PMID- 16707435 TI - Hypoxia increases androgen receptor activity in prostate cancer cells. AB - Recent studies show that prostate cancer cells are able to survive in a hypoxic tumor environment, and the extent of tumor hypoxia correlates with poor clinical outcome. Androgen deprivation, the most common form of prostate cancer therapy, was itself shown to induce a state of transient hypoxia at the microenvironmental level. Because androgen receptor (AR) signaling plays a critical role in prostate cancer, we investigated the effect of hypoxia in regulating AR function. We found that in LNCaP prostate cancer cells, AR binding to the androgen-responsive element (ARE), prostate-specific antigen accumulation, and ARE-reporter gene activity were increased after hypoxia treatment. Hypoxia-enhanced AR function was also observed when AR was exogenously introduced into AR-null DU145 cells. Confocal microscopy and chromatin immunoprecipitation assays showed that AR translocation to the nucleus and AR recruitment to the prostate-specific antigen promoter were facilitated after hypoxia treatment. The AR stimulatory effect seemed to be ligand-dependent because it was abrogated when cells were cultured in an androgen-depleted medium, but was restored with the addition of R1881, a synthetic androgen. The sensitivity of AR activation to R1881 was also increased after hypoxia treatment. Although concentrations of <1 nmol/L R1881 did not induce ARE reporter activity under normoxic conditions, exposure to hypoxia greatly potentiated the AR response to low levels of R1881. Collectively, our results provide compelling evidence that changes in hypoxia/reoxygenation stimulate AR trans-activation and sensitization. The AR-stimulatory effect of an unstable tissue oxygenation milieu of a tumor is likely to contribute to treatment resistance and the emergence of recurrent prostate cancer. PMID- 16707434 TI - Balance between polyoma enhancing activator 3 and activator protein 1 regulates Helicobacter pylori-stimulated matrix metalloproteinase 1 expression. AB - Helicobacter pylori infection and elevated expression of tissue matrix metalloproteinase 1 (MMP-1) are both associated with gastric cancer. We investigated the regulation of MMP-1 expression during H. pylori infection. Real time reverse transcription-PCR was used to examine mucosal MMP-1 mRNA levels in 55 patients with gastric cancers and 61 control patients. Increased MMP-1 mRNA levels in the gastric mucosa and epithelial cells were observed in H. pylori infections in which both the cag pathogenicity island (PAI) and outer inflammatory protein A (OipA) were expressed. The combined induction of c-fos, c jun, and polyoma enhancing activator-3 (pea-3) by H. pylori caused maximal increase in MMP-1 expression. Activation of the MMP-1 promoter by H. pylori involved occupation of the activator protein 1 (AP-1) sites at -72 and -181 and, surprisingly, vacancy of the -88 PEA-3 site. Electrophoretic mobility shift, supershift, and chromatin immunoprecipitation assays showed increased binding of c-Fos and c-Jun to the -72 and -181 AP-1 sites during H. pylori infection. Importantly, during wild-type H. pylori infection, we detected increased PEA-3 binding to the -72AP-1 site and decreased PEA-3 binding to the -88 PEA-3 site. However, during infection with the cag PAI and oipA mutants, PEA-3 binding to the -88 site was detected. MMP-1 and pea-3 activities are increased in gastric cancers. Maximal activation of MMP-1 transcription requires the cag PAI and OipA, which regulate AP-1 and PEA-3 binding. Thus, cag PAI and OipA provide a possible link between bacterial virulence factors and important host factors related to disease pathogenesis. PMID- 16707436 TI - The phosphoinositide 3-kinase/Akt pathway: a new target in human renal cell carcinoma therapy. AB - Metastatic renal cell carcinoma is resistant to current therapies. The phosphoinositide 3-kinase (PI3K)/Akt signaling cascade induces cell growth, cell transformation, and neovascularization. We evaluated whether targeting this pathway could be of therapeutic value against human renal cell carcinoma. The activation of the PI3K/Akt pathway and its role in renal cell carcinoma progression was evaluated in vitro in seven human cell lines by Western blot, cell counting, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, terminal deoxyribonucleotide transferase-mediated nick-end labeling assays, and fluorescence-activated cell sorting analysis, using two PI3K inhibitors, LY294002 and wortmannin, as well as by transfection with various Akt constructs and through Akt knockdown by small interfering RNA (siRNA). In vivo nude mice bearing human renal cell carcinoma tumor xenografts were treated with LY294002 (75 mg/kg/wk, 4 weeks, i.p.). Tumor growth was measured and tumors were subjected to Western blot and immunohistochemical analysis. Akt was constitutively activated in all cell lines. Constitutive phosphorylation of glycogen synthase kinase-3 (GSK-3) was observed in all cell lines, whereas forkhead transcription factor and mammalian target of rapamycin, although expressed, were not constitutively phosphorylated. Exposure to LY294002 or wortmannin decreased Akt activation and GSK-3 phosphorylation and reduced cell growth by up to 70% through induction of cell apoptosis. These effects were confirmed by transfection experiments with Akt constructs or Akt siRNA. Importantly, LY294002 induced up to 50% tumor regression in mice through tumor cell apoptosis. Tumor neovascularization was significantly increased by LY294002 treatment. Blood chemistries showed no adverse effects of the treatment. Our results suggest an important role of PI3K/Akt inhibitors as a potentially useful treatment for patients with renal cell carcinoma. PMID- 16707437 TI - Mirk/Dyrk1b mediates cell survival in rhabdomyosarcomas. AB - Rhabdomyosarcoma is the most common sarcoma in children and is difficult to treat if the primary tumor is nonresectable or if the disease presents with metastases. The function of the serine/threonine kinase Mirk was investigated in this cancer. Mirk has both growth arrest and survival functions in terminally differentiating skeletal myoblasts. Maintenance of Mirk growth arrest properties would cause down regulation of Mirk in transformed myoblasts. Alternatively, Mirk expression would be retained if rhabdomyosarcoma cells used Mirk survival capability. Mirk expression was significant in 12 of 16 clinical cases of rhabdomyosarcoma. Mirk was detected in each rhabdomyosarcoma cell line examined. Mirk was a functional kinase in each of three rhabdomyosarcoma cell lines, where it proved to be more active than in C2C12 skeletal myoblasts. Mirk mediated survival of the majority of clonogenic rhabdomyosarcoma cells. Knockdown of Mirk by RNA interference reduced the fraction of RD and of Rh30 rhabdomyosarcoma cells capable of colony formation 3- to 4-fold in multiple experiments. Depletion of Mirk induced cell death by apoptosis, as shown by increased numbers of terminal deoxynucleotidyl transferase-mediated nick-end labeling-positive cells and by increased binding of Annexin V. Mirk is a stress-activated kinase that mediates expression of contractile proteins in differentiating myoblasts, but Mirk is not essential for muscle formation in the embryo. It is likely that Mirk also facilitates survival of satellite cell-derived rhabdomyoblasts in regenerating skeletal muscle and aids their differentiation. This survival function is maintained in rhabdomyosarcoma, where Mirk may be a novel therapeutic target. PMID- 16707438 TI - Lymph node metastasis in breast cancer xenografts is associated with increased regions of extravascular drain, lymphatic vessel area, and invasive phenotype. AB - Interactions between the tumor stromal compartment and cancer cells play an important role in the spread of cancer. In this study, we have used noninvasive in vivo magnetic resonance imaging (MRI) of two human breast cancer models with significantly different invasiveness, to quantify and understand the role of interstitial fluid transport, lymphatic-convective drain, and vascularization in the regional spread of breast cancer to the axillary lymph nodes. Quantitative fluorescence microscopy was done to morphometrically characterize lymphatic vessels in these tumors. Significant differences in vascular and extravascular transport variables as well as in lymphatic vessel morphology were detected between the two breast cancer models, which also exhibited significant differences in lymph node and lung metastasis. These data are consistent with a role of lymphatic drain in lymph node metastasis and suggest that increased lymph node metastasis may occur due to a combination of increased invasiveness, and reduced extracellular matrix integrity allowing increased pathways of least resistance for the transport of extravascular fluid, as well as tumor cells. It is also possible that lymph node metastasis occurred via the cancer cell-bearing tumoral lymphatic vessels. The congestion of these tumoral lymphatics with cancer cells may have restricted the entry and transport of macromolecules. PMID- 16707439 TI - Stromal antiapoptotic paracrine loop in perineural invasion of prostatic carcinoma. AB - Caveolin-1 (cav-1) is a major scaffolding component of cell membrane invaginations (caveolae). It is involved in sequestering numerous effectors and signaling molecules and has antiapototic activities in prostate cancer. Perineural invasion (PNI) is associated with decreased apoptosis of cancer cells both in human tissues and the in vitro PNI model. We show here that stromal (perineurium) production of cav-1 is involved in a paracrine antiapoptotic loop in PNI. Transforming growth factor-beta1 is up-regulated in the cancer cells as they approach the nerve and is thought to up-regulate cav-1 in the perineurium of nerves with prostate cancer. Cav-1 is then secreted into the microenvironment and used by prostate cancer cells to inhibit apoptosis. In the in vitro PNI model, this phenomenon is partially reversed by neutralizing cav-1 antibodies or using ganglia from cav-1 knockout mice. Our results show a novel paracrine mechanism used by the prostate cancer in PNI to increase their proliferative activity and decrease apoptosis. PMID- 16707440 TI - Membrane-type 4 matrix metalloproteinase promotes breast cancer growth and metastases. AB - Membrane-type matrix metalloproteinases (MT-MMP) constitute a subfamily of six distinct membrane-associated MMPs. Although the contribution of MT1-MMP during different steps of cancer progression has been well documented, the significance of other MT-MMPs is rather unknown. We have investigated the involvement of MT4 MMP, a glycosylphosphatidylinositol-anchored protease, in breast cancer progression. Interestingly, immunohistochemical analysis shows that MT4-MMP production at protein level is strongly increased in epithelial cancer cells of human breast carcinomas compared with normal epithelial cells. Positive staining for MT4-MMP is also detected in lymph node metastases. In contrast, quantitative reverse transcription-PCR analysis reveals similar MT4-MMP mRNA levels in human breast adenocarcinomas and normal breast tissues. Stable transfection of MT4-MMP cDNA in human breast adenocarcinoma MDA-MB-231 cells does not affect in vitro cell proliferation or invasion but strongly promotes primary tumor growth and associated metastases in RAG-1 immunodeficient mice. We provide for the first time evidence that MT4-MMP overproduction accelerates in vivo tumor growth, induces enlargement of i.t. blood vessels, and is associated with increased lung metastases. These results identify MT4-MMP as a new putative target to design anticancer strategies. PMID- 16707441 TI - The S100A4 metastasis factor regulates cellular motility via a direct interaction with myosin-IIA. AB - S100A4, a member of the Ca(2+)-dependent S100 family of proteins, is a metastasis factor that is thought to regulate the motility and invasiveness of cancer cells. Previously, we showed that S100A4 specifically binds to nonmuscle myosin-IIA and promotes the unassembled state. S100A4, thus, provides a connection between the actomyosin cytoskeleton and the regulation of cellular motility; however, the step or steps in the motility cycle that are affected by S100A4 expression have not been investigated. To examine how the biochemical properties of S100A4 affect cell motility, we determined the effect of S100A4 expression on protrusive behavior during chemoattractant-stimulated motility. Our studies show that S100A4 modulates cellular motility by affecting cell polarization, with S100A4 expressing cells displaying few side protrusions and extensive forward protrusions during chemotaxis compared with control cells. To establish a direct link between S100A4 and the regulation of myosin-IIA function, we prepared an antibody to the S100A4 binding site on the myosin-IIA heavy chain that has comparable effects on myosin-IIA assembly as S100A4. Microinjection experiments show that the antibody elicits the same effects on cell polarization as S100A4. Our studies show for the first time that S100A4 promotes directional motility via a direct interaction with myosin-IIA. These findings establish S100A4 as a critical regulator of myosin-II function and metastasis-associated motility. PMID- 16707442 TI - BRCA1 promotes induction of ssDNA by ionizing radiation. AB - The BRCA1 tumor suppressor contributes to the repair of DNA double-strand breaks (DSB) through homologous recombination, but the mechanism is unknown. The rapid accumulation of BRCA1 into nuclear foci in response to induction of DNA breaks suggests that BRCA1 may function in an early step in the repair pathway. We examined the role of BRCA1 in one such early step, the resection of DSBs to generate ssDNA. The appearance of ssDNA in response to ionizing radiation is similar to that of BRCA1 foci formation, suggesting that the two processes are related. Furthermore, BRCA1 colocalizes to ssDNA sites induced by ionizing radiation. Overexpression of BRCA1 causes an increase in cells exhibiting ssDNA induced by ionizing radiation. Mutant BRCA1 that lacks the COOH-terminal BRCT domain also promotes ssDNA but fails to form nuclear foci. Knockdown of BRCA1 expression reduces ssDNA and Rad51 foci formation in response to ionizing radiation. These results indicate that BRCA1 promotes induction of ssDNA in response to ionizing radiation and accumulates at sites of ssDNA. PMID- 16707443 TI - Lack of Rb and p53 delays cerebellar development and predisposes to large cell anaplastic medulloblastoma through amplification of N-Myc and Ptch2. AB - Medulloblastomas are among the most common malignant brain tumors in childhood. They typically arise from neoplastic transformation of granule cell precursors in the cerebellum via deregulation of molecular pathways involved in normal cerebellar development. In a mouse model, we show here that impairment of the balance between proliferation and differentiation of granule cell precursors in the external granular layer of the developing cerebellum predisposes but is not sufficient to induce neoplastic transformation of these progenitor cells. Using array-based chromosomal comparative genomic hybridization, we show that genetic instability resulting from inactivation of the p53 pathway together with deregulation of proliferation induced by Rb loss eventually leads to neoplastic transformation of these cells by acquiring additional genetic mutations, mainly affecting N-Myc and Ptch2 genes. Moreover, we show that p53 loss influences molecular mechanisms that cannot be mimicked by the loss of either p19(ARF), p21, or ATM. PMID- 16707444 TI - Controlled activation of ErbB1/ErbB2 heterodimers promote invasion of three dimensional organized epithelia in an ErbB1-dependent manner: implications for progression of ErbB2-overexpressing tumors. AB - Receptor tyrosine kinases of the ErbB family are implicated in a number of cancers, including that of the breast. ErbB receptors are activated by ligand induced formation of homodimers and heterodimers. Receptor heterodimerization is thought to play a critical role in breast cancers overexpressing multiple members of the ErbB family. Although coexpression of ErbB receptors is associated with poor patient prognosis, the mechanisms by which receptor heterodimerization regulates tumor progression are not clear, due in part to a lack of methods that allow controlled activation of specific receptor heterodimers in mammary epithelial cells. Here, we report an approach to activate ErbB1-ErbB2 heterodimers in a nontumorigenic breast epithelial cell line, MCF-10A, without interference from endogenous ErbB receptors. Using such a method, we show that whereas both ErbB2 homodimers and ErbB1-ErbB2 heterodimers were equally potent in activating the Ras/mitogen-activated protein kinase pathway, the heterodimers were more potent in activating the phosphoinositide 3'-kinase (PI3K) and phospholipase Cgamma1 pathways than ErbB2 homodimers. We combined the dimerization system with a three-dimensional cell culture approach to show that whereas both ErbB2 homodimers and ErbB1-ErbB2 heterodimers induced disruption of three-dimensional acini-like structures, only heterodimers promoted invasion of cells through extracellular matrix. The ability of heterodimers to induce invasion required the ErbB1 kinase activity and required activation of PI3K, Ras/mitogen-activated protein kinase, and phospholipase Cgamma1 signaling pathways. Thus, we have identified cell invasion as a heterodimer-specific biological outcome and suggest that coexpression of ErbB1 may critically regulate invasive progression of ErbB2-positive breast cancers. PMID- 16707445 TI - Malignant transformation of immortalized HaCaT keratinocytes through deregulated nuclear factor kappaB signaling. AB - Previous studies addressing functional aspects of nuclear factor kappaB (NF kappaB) activation in normal and transformed keratinocytes revealed complex and seemingly contradictory roles of this transcription factor in this cell type. In normal skin, NF-kappaB signaling seems to inhibit squamous cell carcinoma development whereas, in squamous cell carcinoma themselves, deregulated NF-kappaB expression and/or signaling is frequently observed. To further investigate this paradox, we focused on NF-kappaB activation as it relates to the transformed phenotype of immortalized but nontumorigenic human keratinocytes (HaCaT cells). We observed that NF-kappaB activity contributed to survival and growth of cultured HaCaT keratinocytes as shown by use of pharmacologic NF-kappaB inhibitors, RNA interference, and inducible overexpression of a dominant interfering IkappaB construct. NF-kappaB activation was largely provided through interaction with extracellular matrix components because preventing cell attachment by forced suspension culture markedly reduced NFkappaB signaling associated with cell death (anoikis); conversely, anoikis was partially reversed by NF-kappaB activation induced either by tumor necrosis factor-alpha treatment or by overexpressing the NF-kappaB p65 subunit in HaCaT cells. Furthermore, overexpression of NF-kappaBp65 in HaCaT cells induced colony formation in soft agar and tumorigenicity in nude mice. In summary, as opposed to normal keratinocytes, immortalized HaCaT keratinocytes provide a cellular context in which deregulated NF-kappaB signaling supports multiple malignant traits in vitro and in vivo. PMID- 16707446 TI - Acid-mediated tumor invasion: a multidisciplinary study. AB - The acid-mediated tumor invasion hypothesis proposes altered glucose metabolism and increased glucose uptake, observed in the vast majority of clinical cancers by fluorodeoxyglucose-positron emission tomography, are critical for development of the invasive phenotype. In this model, increased acid production due to altered glucose metabolism serves as a key intermediate by producing H(+) flow along concentration gradients into adjacent normal tissue. This chronic exposure of peritumoral normal tissue to an acidic microenvironment produces toxicity by: (a) normal cell death caused by the collapse of the transmembrane H(+) gradient inducing necrosis or apoptosis and (b) extracellular matrix degradation through the release of cathepsin B and other proteolytic enzymes. Tumor cells evolve resistance to acid-induced toxicity during carcinogenesis, allowing them to survive and proliferate in low pH microenvironments. This permits them to invade the damaged adjacent normal tissue despite the acid gradients. Here, we describe theoretical and empirical evidence for acid-mediated invasion. In silico simulations using mathematical models provide testable predictions concerning the morphology and cellular and extracellular dynamics at the tumor-host interface. In vivo experiments confirm the presence of peritumoral acid gradients as well as cellular toxicity and extracellular matrix degradation in the normal tissue exposed to the acidic microenvironment. The acid-mediated tumor invasion model provides a simple mechanism linking altered glucose metabolism with the ability of tumor cells to form invasive cancers. PMID- 16707447 TI - Mammary gland tissue targeted overexpression of human protease-activated receptor 1 reveals a novel link to beta-catenin stabilization. AB - Protease-activated receptor 1 (PAR1) is emerging with distinct assignments in tumor biology. We show that tissue targeted overexpression of hPar1 in mice mammary glands results in precocious hyperplasia, characterized by a dense network of ductal side branching and accelerated proliferation. These glands exhibit increased levels of wnt-4 and wnt-7b and a striking beta-catenin stabilization. Nuclear localization of beta-catenin is observed in hPar1 transgenic mouse tissue sections but not in the wild-type, age-matched counterparts. PAR1 induces beta-catenin nuclear localization also in established epithelial tumor cell lines of intact beta-catenin system (transformed on the background of mismatch repair system; RKO cells). We propose hereby that PAR1 mediated beta-catenin stabilization is taking place primarily via the increase of Wnt expression. Enforced expression of a specific Wnt antagonist family member, secreted frizzled receptor protein 5 (SFRP5), efficiently inhibited PAR1-induced beta-catenin stabilization. Likewise, application of either SFRP2 or SFRP5 on epithelial tumor cells completely abrogated PAR1-induced beta-catenin nuclear accumulation. This takes place most likely via inhibition of Wnt signaling at the level of cell surface (forming a neutralizing complex of "Receptors-SFRP-Wnt"). Furthermore, depletion of hPar1 by small interfering RNA (siRNA) vectors markedly inhibited PAR1-induced Wnt-4. The striking stabilization of beta-catenin, inhibited by SFRPs on one hand and Wnt-4 silencing by hPar1 siRNA on the other hand, points to a novel role of hPar1 in Wnt-mediated beta-catenin stabilization. This link between PAR1 and beta-catenin may bear substantial implications both in developmental and tumor progression processes. PMID- 16707448 TI - Earlier onset of tumoral angiogenesis in matrix metalloproteinase-19-deficient mice. AB - Among matrix metalloproteinases (MMP), MMP-19 displays unique structural features and tissue distribution. In contrast to most MMPs, MMP-19 is expressed in normal human epidermis and down-regulated during malignant transformation and dedifferentiation. The contribution of MMP-19 during tumor angiogenesis is presently unknown. In an attempt to give new insights into MMP-19 in vivo functions, angiogenic response of mutant mice lacking MMP-19 was analyzed after transplantation of murine malignant PDVA keratinocytes and after injection of Matrigel supplemented with basic fibroblast growth factor. In situ hybridization and immunohistochemical analysis revealed that MMP-19 is produced by host mesenchymal cells but not by endothelial capillary cells or CD11b-positive inflammatory cells. Based on a new computer-assisted method of quantification, we provide evidence that host MMP-19 deficiency was associated with an increased early angiogenic response. In addition, increased tumor invasion was observed in MMP-19-/- mice. We conclude that, in contrast to most MMPs that promote tumor progression, MMP-19 is a negative regulator of early steps of tumor angiogenesis and invasion. These data highlight the requirement to understand the individual functions of each MMP to improve anticancer strategies. PMID- 16707449 TI - Tumor cell-derived and macrophage-derived cathepsin B promotes progression and lung metastasis of mammary cancer. AB - Proteolysis in close vicinity of tumor cells is a hallmark of cancer invasion and metastasis. We show here that mouse mammary tumor virus-polyoma middle T antigen (PyMT) transgenic mice deficient for the cysteine protease cathepsin B (CTSB) exhibited a significantly delayed onset and reduced growth rate of mammary cancers compared with wild-type PyMT mice. Lung metastasis volumes were significantly reduced in PyMT;ctsb(+/-), an effect that was not further enhanced in PyMT;ctsb(-/-) mice. Furthermore, lung colonization studies of PyMT cells with different CTSB genotypes injected into congenic wild-type mice and in vitro Matrigel invasion assays confirmed a specific role for tumor-derived CTSB in invasion and metastasis. Interestingly, cell surface labeling of cysteine cathepsins by the active site probe DCG-04 detected up-regulation of cathepsin X on PyMT;ctsb(-/-) cells. Treatment of cells with a neutralizing anti-cathepsin X antibody significantly reduced Matrigel invasion of PyMT;ctsb(-/-) cells but did not affect invasion of PyMT;ctsb(+/+) or PyMT;ctsb(+/-) cells, indicating a compensatory function of cathepsin X in CTSB-deficient tumor cells. Finally, an adoptive transfer model, in which ctsb(+/+), ctsb(+/-), and ctsb(-/-) recipient mice were challenged with PyMT;ctsb(+/+) cells, was used to address the role of stroma-derived CTSB in lung metastasis formation. Notably, ctsb(-/-) mice showed reduced number and volume of lung colonies, and infiltrating macrophages showed a strongly up-regulated expression of CTSB within metastatic cell populations. These results indicate that both cancer cell-derived and stroma cell-derived (i.e., macrophages) CTSB plays an important role in tumor progression and metastasis. PMID- 16707451 TI - Regulation of microtubule-dependent protein transport by the TSC2/mammalian target of rapamycin pathway. AB - Protein transport plays a critical role in the interaction of the cell with its environment. Recent studies have identified TSC1 and TSC2, two tumor suppressor genes involved in tuberous sclerosis complex, as regulators of the mammalian target of rapamycin (mTOR) pathway. Cells deficient in TSC1 or TSC2 possess high levels of Rheb-GTP resulting in constitutive mTOR activation. We have shown previously that the TSC1/TSC2 complex is involved in post-Golgi transport of VSVG and caveolin-1 in mammalian cells. Here, we show that modulation of mTOR activity affects caveolin-1 localization and that this effect is independent of p70S6K. Tsc1- and Tsc2-null cells exhibit abnormal caveolin-1 localization that is accompanied by disorganized microtubules in the subcortical region. Analyses of green fluorescent protein-EB1 and tubulin in live mutant cells suggest a failure of the plus-ends to sense cortical signals and to halt microtubule growth. Down regulation of CLIP-170, a putative mTOR substrate with microtubule-binding properties, rescued the abnormal microtubule arrangement and caveolin-1 localization in Tsc2-/- cells. Together, these findings highlight a novel role of the TSC2/mTOR pathway in regulating microtubule-dependent protein transport. PMID- 16707450 TI - Tight junction protein claudin-1 enhances the invasive activity of oral squamous cell carcinoma cells by promoting cleavage of laminin-5 gamma2 chain via matrix metalloproteinase (MMP)-2 and membrane-type MMP-1. AB - Although adherent junctions have been extensively studied, the role of tight junctions in cancer cell invasion is not sufficiently explored. We investigated whether claudin-1, a component of tight junctions, regulated invasion activity in oral squamous cell carcinoma (OSC) cells. The expression of claudin-1, activity of matrix metalloproteinase (MMP)-2, and cleavage of laminin-5 gamma2 chains were assessed by Western blot analysis, immunohistochemistry, and zymography in OSC cell lines (OSC-4 and NOS-2, highly invasive; OSC-7, weakly invasive) and their xenografts in severe combined immunodeficient (SCID) mice. The influence of claudin-1 small interfering RNA (siRNA) on the invasion activity of the cell lines was also investigated. Compared with OSC-7, both OSC-4 and NOS-2 more strongly expressed claudin-1 and possessed high activities of MMP-2 and MMP-9. Tumors formed in the tongues of SCID mice xenografted with OSC-4, NOS-2, and OSC 7 immunohistochemically revealed strong, moderate, and weak expression of laminin 5 gamma2 chains, respectively, and laminin-5 gamma2 chains were secreted in the conditioned medium of the cancer cells in parallel with the in vivo results. Claudin-1 siRNA largely suppressed the invasion of OSC-4 and decreased the activation of MMP-2, the expression of membrane-type MMP-1 (MT1-MMP), and the cleavage of laminin-5 gamma2. In addition, not only antibodies against MT1-MMP and epidermal growth factor receptor (EGFR) but also MMP-2 and EGFR inhibitors strongly suppressed the invasion activity of OSC-4. These results suggest that claudin-1 up-regulates cancer cell invasion activity through activation of MT1 MMP and MMP-2, which results in enhanced cleavage of laminin-5 gamma2 chains. PMID- 16707452 TI - Selective expansion of marginal zone B cells in Emicro-API2-MALT1 mice is linked to enhanced IkappaB kinase gamma polyubiquitination. AB - The translocation t(11;18)(q21;q21) that generates an API2-MALT1 fusion protein is the most common structural abnormality among the genetic defects reported in mucosa-associated lymphoid tissue (MALT)-type lymphomas, and its presence correlates with the apparent lack of further genetic instability or chromosomal imbalances. Hence, constitutive nuclear factor-kappaB (NF-kappaB) activation induced by the API2-MALT1 fusion protein is considered essential for B-cell transformation. To examine its role in B-cell development and lymphomagenesis, Emu-API2-MALT1 transgenic mice were produced. Our data show that expression of the API2-MALT1 fusion protein alone is not sufficient for the development of lymphoma masses within 50 weeks. Nevertheless, API2-MALT1 expression affected B cell maturation in the bone marrow and triggered the specific expansion of splenic marginal zone B cells. Polyubiquitination of IkappaB kinase gamma (IKKgamma), indicative for enhanced NF-kappaB activation, was increased in splenic lymphocytes and promoted the survival of B cells ex vivo. In addition, we show that the API2-MALT1 fusion resided in the cholesterol- and sphingolipid enriched membrane microdomains, termed lipid rafts. We provide evidence that association of the MALT1 COOH terminal with the lipid rafts, which is mediated by the API2 portion, is sufficient to trigger NF-kappaB activation via enhanced polyubiquitination of IKKgamma. Taken together, these data support the hypothesis that the API2-MALT1 fusion protein can contribute to MALT lymphoma formation via increased NF-kappaB activation. PMID- 16707453 TI - Progression-specific genes identified by expression profiling of matched ductal carcinomas in situ and invasive breast tumors, combining laser capture microdissection and oligonucleotide microarray analysis. AB - Becoming invasive is a crucial step in breast cancer oncogenesis. At this point, a lesion carries the potential for spreading and metastasis--a process, whose molecular characteristics still remain poorly understood. In this article, we describe a matched-pair analysis of ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC) of nine breast ductal carcinomas to identify novel molecular markers characterizing the transition from DCIS to IDC. The purpose of this study was to better understand the molecular biology of this transition and to identify candidate genes whose products might serve as prognostic markers and/or as molecular targets for treatment. To obtain cellular-based gene expression profiles from epithelial tumor cells, we combined laser capture microdissection with a T7-based two-round RNA amplification and Affymetrix oligonucleotide microarray analysis. Altogether, a set of 24 tumor samples was analyzed, comprised of nine matched DCIS/IDC and replicate DCIS/IDC preparations from three of the nine tumors. Cluster analysis on expression data shows the robustness and reproducibility of the techniques we established. Using multiple statistical methods, 546 significantly differentially expressed probe sets were identified. Eighteen candidate genes were evaluated by RT-PCR. Examples of genes already known to be associated with breast cancer invasion are BPAG1, LRRC15, MMP11, and PLAU. The expression of BPAG1, DACT1, GREM1, MEF2C, SART2, and TNFAIP6 was localized to epithelial tumor cells by in situ hybridization and/or immunohistochemistry, confirming the accuracy of laser capture microdissection sampling and microarray analysis. PMID- 16707454 TI - Acetyl-CoA carboxylase alpha is essential to breast cancer cell survival. AB - Activation of de novo fatty acid synthesis is a characteristic feature of cancer cells. We have recently described an interaction between acetyl-CoA carboxylase alpha (ACCalpha), a key enzyme in fatty acid synthesis, and BRCA1, which indicates a possible connection between lipid synthesis and genetic factors involved in susceptibility to breast and ovarian cancers. For this reason, we explored the role of ACCalpha in breast cancer cell survival using an RNA interference (RNAi) approach. We show that specific silencing of either the ACCalpha or the fatty acid synthase (FAS) genes in cancer cells results in a major decrease in palmitic acid synthesis. Depletion of the cellular pool of palmitic acid is associated with induction of apoptosis concomitant with the formation of reactive oxygen species (ROS) and mitochondrial impairment. Expression of a small interfering RNA (siRNA)-resistant form of ACCalpha mRNA prevented the effect of ACCalpha-RNAi but failed to prevent the effect of FAS gene silencing. Furthermore, supplementation of the culture medium with palmitate or with the antioxidant vitamin E resulted in the complete rescue of cells from both ACCalpha and FAS siRNA-induced apoptosis. Finally, human mammary epithelial cells are resistant to RNAi against either ACCalpha or FAS. These data confirm the importance of lipogenesis in cancer cell survival and indicate that this pathway represents a key target for antineoplastic therapy that, however, might require specific dietary recommendation for full efficacy. PMID- 16707455 TI - Protein 4.1B/differentially expressed in adenocarcinoma of the lung-1 functions as a growth suppressor in meningioma cells by activating Rac1-dependent c-Jun NH(2)-kinase signaling. AB - Meningiomas are the second most common brain tumor in adults, yet comparatively little is presently known about the dysregulated growth control pathways involved in their formation and progression. One of the most frequently observed genetic changes in benign meningioma involves loss of protein 4.1B expression. Previous studies from our laboratory have shown that protein 4.1B growth suppression in meningioma is associated with the activation of the c-Jun-NH(2)-kinase (JNK) pathway and requires localization of a small unique region (U2 domain) of protein 4.1B to the plasma membrane. To define the relationship between protein 4.1B expression and JNK activation, as well as to determine the mechanism of JNK activation by protein 4.1B, we used a combination of genetic and pharmacologic approaches. In this report, we show that protein 4.1B/differentially expressed in adenocarcinoma of the lung-1 (DAL-1) expression in meningioma cells in vitro results in JNK activation, which requires the sequential activation of Src, Rac1, and JNK. In addition, inhibition of Rac1 or JNK activation abrogates protein 4.1B/DAL-1 growth suppression and cyclin A regulation. Last, protein 4.1B/DAL-1 regulation of this critical growth control pathway in meningioma cells requires the presence of the U2 domain. Collectively, these observations provide the first mechanistic insights into the function of protein 4.1B as a growth regulator in meningioma cells. PMID- 16707456 TI - Epidermal growth factor induces insulin receptor substrate-2 in breast cancer cells via c-Jun NH(2)-terminal kinase/activator protein-1 signaling to regulate cell migration. AB - The epidermal growth factor (EGF) and insulin-like growth factor (IGF) signaling pathways are critically involved in cancer development and progression. However, how these two signals cross-talk with each other to regulate cancer cell growth is not clearly understood. In this study, we found that EGF remarkably induced expression of major IGF signaling components, insulin receptor substrate (IRS)-1 and IRS-2, an effect that could be blocked by EGF receptor (EGFR) tyrosine kinase inhibitors. Although both extracellular signal-regulated kinase and c-Jun NH(2) terminal kinase (JNK) signaling pathways were involved in the EGF up-regulation of IRS-1, the IRS-2 induction by EGF was specifically mediated by JNK signaling. Consistent with this, EGF increased IRS-2 promoter activity, which was associated with recruitment of activator protein-1 (AP-1) transcription factors and was inhibited by blocking AP-1 activity. Moreover, EGF treatment enhanced IGF-I and integrin engagement-elicited tyrosine phosphorylation of IRS and their downstream signaling, such as binding to phosphatidylinositol 3'-kinase regulatory subunit p85. Finally, repressing the induction of IRS-2 levels abolished the EGF enhancement of cell motility, suggesting that increased IRS-2 is essential for the EGF regulation of breast cancer cell migration. Taken together, our results reveal a novel mechanism of cross-talk between the EGF and IGF signaling pathways, which could have implications in therapeutic applications of targeting EGFR in tumors. Because AP-1 activity is involved in breast cancer progression, our work may also suggest IRS-2 as a useful marker for aggressive breast cancer. PMID- 16707457 TI - Keratoepithelin suppresses the progression of experimental human neuroblastomas. AB - Neuroblastoma is the most common extracranial childhood tumor. High expression of activin A is associated with a favorable prognosis, but the contributing mechanisms have remained unclear. Our previous demonstration of the activin A mediated up-regulation of keratoepithelin led to the consideration that keratoepithelin could modulate neuroblastoma growth and/or progression. We report here that enhanced keratoepithelin expression in human neuroblastoma cells suppresses neuroblastoma cell cohesion and adhesion to various extracellular matrix proteins and that it inhibits neuroblastoma cell proliferation and invasion in vitro and in vivo. Using microarray analysis, we identified several keratoepithelin-regulated genes that may contribute to these biological changes. Together with the observation that keratoepithelin is expressed in human neuroblastomas in vivo, our data suggest that keratoepithelin could play a beneficial role in neuroblastoma development and/or progression. PMID- 16707458 TI - Microarray-assisted pathway analysis identifies mitogen-activated protein kinase signaling as a mediator of resistance to the green tea polyphenol epigallocatechin 3-gallate in her-2/neu-overexpressing breast cancer cells. AB - Overexpression of the epidermal growth factor receptor family member Her-2/neu in breast cancer leads to autophosphorylation of the receptor and induction of multiple downstream signaling pathways, including the Akt kinase to nuclear factor-kappaB (NF-kappaB) cascade that is associated with poor prognosis. Previously, we showed that the green tea polyphenol epigallocatechin 3-gallate (EGCG) inhibits growth of NF639 Her-2/neu-driven breast cancer cells via reducing receptor autophosphorylation and downstream Akt and NF-kappaB activities. Interestingly, upon prolonged culture in the presence of EGCG, cells resistant to the polyphenol could be isolated. Here, we report that resistant cells have lost tyrosine phosphorylation on the Her-2/neu receptor. Surprisingly, they displayed elevated NF-kappaB activity, and inhibition of this activity sensitized cells to EGCG. Data from microarray studies of the original and resistant NF639 populations of cells were subjected to Gene Set Enrichment Analysis pathway assessment, which revealed that the mitogen activated protein kinase (MAPK) pathway was activated in the resistant cells. Treatment of the resistant cells with the MAPK inhibitor U0216 reduced growth in soft agar and invasive phenotype, whereas the combination of EGCG and U0216 resulted in cells with a cobblestone epithelial phenotype. Thus, activation of the MAPK pathway mediates resistance to EGCG. PMID- 16707459 TI - The c-Myc oncogene directly induces the H19 noncoding RNA by allele-specific binding to potentiate tumorigenesis. AB - The product of the MYC oncogene is widely deregulated in cancer and functions as a regulator of gene transcription. Despite an extensive profile of regulated genes, the transcriptional targets of c-Myc essential for transformation remain unclear. In this study, we show that c-Myc significantly induces the expression of the H19 noncoding RNA in diverse cell types, including breast epithelial, glioblastoma, and fibroblast cells. c-Myc binds to evolutionarily conserved E boxes near the imprinting control region to facilitate histone acetylation and transcriptional initiation of the H19 promoter. In addition, c-Myc down-regulates the expression of insulin-like growth factor 2 (IGF2), the reciprocally imprinted gene at the H19/IGF2 locus. We show that c-Myc regulates these two genes independently and does not affect H19 imprinting. Indeed, allele-specific chromatin immunoprecipitation and expression analyses indicate that c-Myc binds and drives the expression of only the maternal H19 allele. The role of H19 in transformation is addressed using a knockdown approach and shows that down regulation of H19 significantly decreases breast and lung cancer cell clonogenicity and anchorage-independent growth. In addition, c-Myc and H19 expression shows strong association in primary breast and lung carcinomas. This work indicates that c-Myc induction of the H19 gene product holds an important role in transformation. PMID- 16707460 TI - Cyclooxygenase-2-dependent regulation of E-cadherin: prostaglandin E(2) induces transcriptional repressors ZEB1 and snail in non-small cell lung cancer. AB - Elevated tumor cyclooxygenase-2 (COX-2) expression is associated with tumor invasion, metastasis, and poor prognosis in non-small cell lung cancer (NSCLC). Here, we report that COX-2-dependent pathways contribute to the modulation of E cadherin expression in NSCLC. First, whereas genetically modified COX-2-sense (COX-2-S) NSCLC cells expressed low E-cadherin and showed diminished capacity for cellular aggregation, genetic or pharmacologic inhibition of tumor COX-2 led to increased E-cadherin expression and resulted in augmented homotypic cellular aggregation among NSCLC cells in vitro. An inverse relationship between COX-2 and E-cadherin was shown in situ by double immunohistochemical staining of human lung adenocarcinoma tissue sections. Second, treatment of NSCLC cells with exogenous prostaglandin E(2) (PGE(2)) significantly decreased the expression of E-cadherin, whereas treatment of COX-2-S cells with celecoxib (1 mumol/L) led to increased E cadherin expression. Third, the transcriptional suppressors of E-cadherin, ZEB1 and Snail, were up-regulated in COX-2-S cells or PGE(2)-treated NSCLC cells but decreased in COX-2-antisense cells. PGE(2) exposure led to enhanced ZEB1 and Snail binding at the chromatin level as determined by chromatin immunoprecipitation assays. Small interfering RNA-mediated knockdown of ZEB1 or Snail interrupted the capacity of PGE(2) to down-regulate E-cadherin. Fourth, an inverse relationship between E-cadherin and ZEB1 and a direct relationship between COX-2 and ZEB1 were shown by immunohistochemical staining of human lung adenocarcinoma tissue sections. These findings indicate that PGE(2), in autocrine or paracrine fashion, modulates transcriptional repressors of E-cadherin and thereby regulates COX-2-dependent E-cadherin expression in NSCLC. Thus, blocking PGE(2) production or activity may contribute to both prevention and treatment of NSCLC. PMID- 16707461 TI - PTEN/Akt signaling through epidermal growth factor receptor is prerequisite for angiogenesis by hepatocellular carcinoma cells that is susceptible to inhibition by gefitinib. AB - Hepatocellular carcinoma (HCC) is one of the most common tumor-related causes of death worldwide for which there is still no satisfactory treatment. We previously reported the antiangiogenic effect of gefitinib, a selective epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor that has been used successfully to treat lung cancer. In this study, we investigated the effects of gefitinib on tumor-induced angiogenesis by using HCC cell lines (HCC3, CBO12C3, and AD3) in vitro as well as in vivo. Oral administration of gefitinib inhibited angiogenesis induced by HCC3 and CBO12C3, but not by AD3 in the mouse dorsal air sac model. Production of both vascular endothelial growth factor (VEGF) and chemokine C-X-C motif ligand 1 (CXCL1) by EGF-stimulated HCC was more markedly inhibited by gefitinib in HCC3 and CBO12C3 cells than in AD3 cells. EGF stimulated the phosphorylation of EGFR, Akt, and extracellular signal-regulated kinase 1/2 (ERK1/2) in HCC3 and CBO12C3 cells, whereas EGF stimulated phosphorylation of EGFR and ERK1/2, but not Akt in AD3 cells. In fact, Akt was constitutively activated in the absence of EGF in AD3 cells. Gefitinib inhibited Akt phosphorylation in all three cell lines, but it was about five times less effective in AD3 cells. The concentration of PTEN in AD3 cells was about a half that in HCC3 and CBO12C3 cells. Transfection of HCC3 cells with PTEN small interfering RNA reduced their sensitivity to gefitinib in terms of its inhibitory effect on both Akt phosphorylation and the production of VEGF and CXCL1. In conclusion, effect of gefitinib on HCC-induced angiogenesis depends on its inhibition of the production of angiogenic factors, probably involving a PTEN/Akt signaling pathway. PMID- 16707462 TI - Preclinical evaluation of a potent novel DNA-dependent protein kinase inhibitor NU7441. AB - DNA double-strand breaks (DSB) are the most cytotoxic lesions induced by ionizing radiation and topoisomerase II poisons, such as etoposide and doxorubicin. A major pathway for the repair of DSB is nonhomologous end joining, which requires DNA-dependent protein kinase (DNA-PK) activity. We investigated the therapeutic use of a potent, specific DNA-PK inhibitor (NU7441) in models of human cancer. We measured chemosensitization by NU7441 of topoisomerase II poisons and radiosensitization in cells deficient and proficient in DNA-PK(CS) (V3 and V3 YAC) and p53 wild type (LoVo) and p53 mutant (SW620) human colon cancer cell lines by clonogenic survival assay. Effects of NU7441 on DSB repair and cell cycle arrest were measured by gammaH2AX foci and flow cytometry. Tissue distribution of NU7441 and potentiation of etoposide activity were determined in mice bearing SW620 tumors. NU7441 increased the cytotoxicity of ionizing radiation and etoposide in SW620, LoVo, and V3-YAC cells but not in V3 cells, confirming that potentiation was due to DNA-PK inhibition. NU7441 substantially retarded the repair of ionizing radiation-induced and etoposide-induced DSB. NU7441 appreciably increased G(2)-M accumulation induced by ionizing radiation, etoposide, and doxorubicin in both SW620 and LoVo cells. In mice bearing SW620 xenografts, NU7441 concentrations in the tumor necessary for chemopotentiation in vitro were maintained for at least 4 hours at nontoxic doses. NU7441 increased etoposide-induced tumor growth delay 2-fold without exacerbating etoposide toxicity to unacceptable levels. In conclusion, NU7441 shows sufficient proof of principle through in vitro and in vivo chemosensitization and radiosensitization to justify further development of DNA-PK inhibitors for clinical use. PMID- 16707463 TI - Apo2l/Tumor necrosis factor-related apoptosis-inducing ligand prevents breast cancer-induced bone destruction in a mouse model. AB - Breast cancer is the most common carcinoma that metastasizes to bone. To examine the efficacy of recombinant soluble Apo2 ligand (Apo2L)/tumor necrosis factor related apoptosis-inducing ligand (TRAIL) against breast cancer growth in bone, we established a mouse model in which MDA-MB-231 human breast cancer cells were transplanted directly into the marrow cavity of the tibiae of athymic nude mice producing osteolytic lesions in the area of injection. All vehicle-treated control animals developed large lesions that established in the marrow cavity, eroded the cortical bone, and invaded the surrounding soft tissue, as assessed by radiography, micro-computed tomography, and histology. In contrast, animals treated with recombinant soluble Apo2L/TRAIL showed significant conservation of the tibiae, with 85% reduction in osteolysis, 90% reduction in tumor burden, and no detectable soft tissue invasion. Tumor cells explanted from Apo2L/TRAIL treated animals were significantly more resistant to the effects of Apo2L/TRAIL when compared with the cells explanted from the vehicle-treated control animals, suggesting that prolonged treatment with Apo2/TRAIL in vivo selects for a resistant phenotype. However, such resistance was readily reversed when Apo2L/TRAIL was used in combination with clinically relevant chemotherapeutic drugs, including taxol, etoposide, doxorubicin, cisplatin, or the histone deacetylase inhibitor suberoylanilide hydroxamic acid. These studies show for the first time that Apo2L/TRAIL can prevent breast cancer-induced bone destruction and highlight the potential of this ligand for the treatment of metastatic breast cancer in bone. PMID- 16707464 TI - Inhibition of tumor growth and elimination of multiple metastases in human prostate and breast xenografts by systemic inoculation of a host defense-like lytic peptide. AB - We report on a short host defense-like peptide that targets and arrests the growth of aggressive and hormone-resistant primary human prostate and breast tumors and prevents their experimental and spontaneous metastases, respectively, when systemically inoculated to immunodeficient mice. These effects are correlated with increased necrosis of the tumor cells and a significant decrease in the overall tumor microvessel density, as well as newly formed capillary tubes and prostate-specific antigen secretion (in prostate tumors). Growth inhibition of orthotopic tumors derived from stably transfected highly fluorescent human breast cancer cells and prevention of their naturally occurring metastases were visualized in real time by using noninvasive whole-body optical imaging. The exclusive selectivity of the peptide towards cancer derives from its specific binding to surface phosphatidylserine and the killing of the cancer cells via cytoplasmic membrane depolarization. These data indicate that membrane disruption can provide a therapeutic means of inhibiting tumor growth and preventing metastases of various cancers. PMID- 16707465 TI - c-Jun NH(2)-terminal kinase signaling axis regulates diallyl trisulfide-induced generation of reactive oxygen species and cell cycle arrest in human prostate cancer cells. AB - We have shown previously that generation of reactive oxygen species (ROS) is a critical event in G(2)-M phase cell cycle arrest caused by diallyl trisulfide (DATS), which is a highly promising anticancer constituent of processed garlic. Using DU145 and PC-3 human prostate cancer cells as a model, we now report a novel mechanism involving c-Jun NH(2)-terminal kinase (JNK) signaling axis, which is known for its role in regulation of cell survival and apoptosis, in DATS induced ROS production. The DATS-induced ROS generation, G(2)-M phase cell cycle arrest and degradation, and hyperphosphorylation of Cdc25C were significantly attenuated in the presence of EUK134, a combined mimetic of superoxide dismutase and catalase. Interestingly, the DATS-induced ROS generation and G(2)-M phase cell cycle arrest were also inhibited significantly in the presence of desferrioxamine, an iron chelator, but this protection was not observed with iron saturated desferrioxamine. DATS treatment caused a marked increase in the level of labile iron that was accompanied by degradation of light chain of iron storage protein ferritin. Interestingly, DATS-mediated degradation of ferritin, increase in labile iron pool, ROS generation, and/or cell cycle arrest were significantly attenuated by ectopic expression of a catalytically inactive mutant of JNK kinase 2 and RNA interference of stress-activated protein kinase/extracellular signal regulated kinase 1 (SEK1), upstream kinases in JNK signal transduction pathway. In conclusion, the present study provides experimental evidence to indicate existence of a novel pathway involving JNK signaling axis in regulation of DATS induced ROS generation. PMID- 16707466 TI - Resistance to imatinib of bcr/abl p190 lymphoblastic leukemia cells. AB - Around 20% of patients with acute lymphoblastic leukemia are Philadelphia chromosome positive (Ph-positive acute lymphoblastic leukemia) and express the Bcr/Abl tyrosine kinase. Treatment with the tyrosine kinase inhibitor Imatinib is currently standard for chronic myelogenous leukemia, which is also caused by Bcr/Abl. However, Imatinib has shown limited efficacy for treating Ph-positive acute lymphoblastic leukemia. In our study, we have investigated the effect of Imatinib therapy on murine P190 Bcr/Abl lymphoblastic leukemia cells. Three of four cultures were very sensitive to treatment with 5 mumol/L Imatinib. Significant cell death also initially occurred when the same cultures were treated in the presence of stromal support. However, after 6 days, remaining cells started to proliferate vigorously. The Bcr/Abl tyrosine kinase present in the cells that were now able to multiply in the presence of 5 mumol/L Imatinib was still inhibited by the drug. In concordance with this, the Abl ATP-binding pocket domain of Bcr/Abl in the resistant cells did not contain point mutations which would make the protein Imatinib resistant. The effect of stroma in selecting Imatinib-resistant lymphoblasts did not require direct cell-cell contact. SDF-1alpha could substitute for the presence of stromal cells. Our results show that stroma selects Imatinib-resistant Bcr/Abl P190 lymphoblasts that are less dependent on Bcr/Abl tyrosine kinase activity. Therefore, therapy for Ph-positive acute lymphoblastic leukemia, aimed at interfering with the protective effect of stroma in combination with Imatinib, could be of benefit for the eradication of the leukemic cells. PMID- 16707467 TI - Potent in vivo anti-breast cancer activity of IN-2001, a novel inhibitor of histone deacetylase, in MMTV/c-Neu mice. AB - A novel synthetic inhibitor of histone deacetylase (HDAC), 3-(4 dimethylaminophenyl)-N-hydroxy-2-propenamide (IN-2001), was examined for its antitumor activity and for the underlying molecular mechanisms of any such activity. IN-2001 effectively inhibited cellular HDAC activity (IC(50), 5.42 nmol/L) in MCF-7 human breast cancer cells. Based on the Western blot analysis, this HDAC inhibitory effect of IN-2001 was confirmed by an increase in histone H4 acetylation from the IN-2001-treated breast cancer cells. IN-2001 suppressed mammary tumor growth in MMTV/c-Neu transgenic mice and also showed higher apoptotic index and lower lymphatic invasion compared with controls. In human breast cancer cells (MCF-7, T47D, MDA-MB-231, and MDA-MB-468), IN-2001 induced cell cycle arrest at G(2)-M phase through up-regulation of p21(WAF1) and p27(KIP1) and eventually caused apoptosis. IN-2001-induced apoptosis was caspase dependent and seems mediated through an increase in Bax/Bcl-2 ratio. Taken together, our data indicate that this novel HDAC inhibitor is a promising therapeutic agent against human breast cancer. PMID- 16707468 TI - KRAS(D13) Promotes apoptosis of human colorectal tumor cells by ReovirusT3D and oxaliplatin but not by tumor necrosis factor-related apoptosis-inducing ligand. AB - Colorectal tumors frequently contain activating mutations in KRAS. ReovirusT3D is an oncolytic virus that preferentially kills tumor cells with an activated Ras pathway. Here we have assessed the contribution of endogenous mutant KRAS in human colorectal cancer cell lines to ReovirusT3D replication and to tumor cell oncolysis. In addition, treatment combinations involving ReovirusT3D, oxaliplatin, and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) were tested for their efficacy in tumor cell killing. The mutation status of KRAS did not predict the sensitivity of a panel of human colorectal cancer cell lines to ReovirusT3D. Virus replication was observed in all cell lines tested regardless of KRAS status and was not affected by deletion of endogenous mutant KRAS(D13). However, deletion of KRAS(D13) or p53 did reduce apoptosis induction by ReovirusT3D whereas deletion of beta-catenin(DeltaS45) had no effect. Likewise, KRAS(D13)- or p53-deficient cells display reduced sensitivity to oxaliplatin but not to death receptor activation by TRAIL. Finally, the treatment of colorectal cancer cells with ReovirusT3D combined with either oxaliplatin or TRAIL resulted in a nonsynergistic increase in tumor cell killing. We conclude that oncolysis of human tumor cells by ReovirusT3D is not determined by the extent of virus replication but by their sensitivity to apoptosis induction. Oncogenic KRAS(D13) increases tumor cell sensitivity to activation of the cell intrinsic apoptosis pathway without affecting ReovirusT3D replication. PMID- 16707469 TI - Histone deacetylase inhibitors suppress the inducibility of nuclear factor-kappaB by tumor necrosis factor-alpha receptor-1 down-regulation. AB - Recently, the inhibition of histone deacetylase (HDAC) enzymes has attracted attention in the oncologic community as a new therapeutic opportunity for hematologic and solid tumors including non-small cell lung cancer (NSCLC). In hematologic malignancies, such as diffuse large B-cell lymphoma, the HDAC inhibitor (HDI), suberoylanilide hydroxamic acid (SAHA), has recently entered phase II and III clinical trials. To further advance our understanding of their action on tumor cells, we investigated the possible effect of HDI treatment on the functionality of the nuclear factor-kappaB (NF-kappaB) pathway in NSCLC. We found that in the NSCLC cell lines, A549 and NCI-H460, the NF-kappaB pathway was strongly inducible, for example, by stimulation with tumor necrosis factor-alpha (TNF-alpha). Incubation of several NSCLC cell lines with HDIs resulted in greatly reduced gene expression of TNF-alpha receptor-1. HDI-treated A549 and NCI-H460 cells down-regulated TNF-alpha receptor-1 mRNA and protein levels as well as surface exposure, and consequently responded to TNF-alpha treatment with reduced IKK phosphorylation and activation, delayed IkappaB-alpha phosphorylation, and attenuated NF-kappaB nuclear translocation and DNA binding. Accordingly, stimulation of NF-kappaB target gene expression by TNF-alpha was strongly decreased. In addition, we observed that SAHA displayed antitumor efficacy in vivo against A549 xenografts grown on nude mice. HDIs, therefore, might beneficially contribute to tumor treatment, possibly by reducing the responsiveness of tumor cells to the TNF-alpha-mediated activation of the NF kappaB pathway. These findings also hint at a possible use of HDIs in inflammatory diseases, which are associated with the overproduction of TNF-alpha, such as rheumatoid arthritis or Crohn's disease. PMID- 16707470 TI - Doxorubicin plus interleukin-2 chemoimmunotherapy against breast cancer in mice. AB - As recently characterized, following s.c. implantation into syngeneic C57BL/6 mice, E0771 tumor invades locally into dermal layers and peritoneum, metastasizes to the lung, and induces a nonspecific immunosuppression in the host. Using this breast medullar adenocarcinoma model, a therapy consisting of a single moderate dose of doxorubicin followed by twice daily moderate doses of interleukin-2 for 30 days was examined for efficacy and mechanism of action when given to animals with established disease. This combination treatment, but not combinants alone, resulted in tumor-free long-term survival of 40% of the mice without significant toxicity and 83% of survivors had immune memory specific for E0771 cells. Treatment also decreased immune suppression induced by E0771 tumor. Full response to treatment required functional CD8(+) T cells, whereas depletion of natural killer cells caused only a reduction in response rate. A serum "biomarker" profile that correlated with, and seemed predictive of, response to treatment was identified by nuclear magnetic resonance-based metabonomic analysis. The efficacy of this nontoxic treatment and the potential to be able to predict which individual is responding to treatment are characteristics that make this chemoimmunotherapy attractive for clinical testing. PMID- 16707471 TI - Therapeutic efficacy of antigen-specific vaccination and toll-like receptor stimulation against established transplanted and autochthonous melanoma in mice. AB - Malignant melanoma is an attractive model disease for the development of antigen specific immunotherapy because many antigens recognized by tumor-specific T cells have been identified. In C57BL/6 mice, genetic immunization with recombinant adenovirus encoding xenogeneic human tyrosinase-related protein 2 (Ad-hTRP2) induces protective but not therapeutic cellular immunity against growth of transplanted B16 melanoma cells. Here, we additionally applied CpG DNA and synthetic double-stranded RNA, which activate the innate immune system via Toll like receptors (TLR). Both adenoviral vaccination and peritumoral injections of TLR ligands were required for rejection of established B16 melanoma in the skin. To more closely mimic the clinical situation in patients with melanoma, we evaluated this combined immunotherapeutic strategy in genetically modified mice, which overexpress hepatocyte growth factor (HGF) and carry an oncogenic mutation in the cyclin-dependent kinase 4 (CDK4)(R24C). HGF x CDK4(R24C) mice rapidly develop multiple invasive melanomas in the skin following neonatal carcinogen treatment, which spontaneously metastasize to lymph nodes and lungs. Vaccination with Ad-hTRP2 followed by injections of TLR ligands resulted in delayed growth of autochthonous primary melanomas in the skin and reduction in the number of spontaneous lung metastases but did not induce tumor regression. Carcinogen treated HGF x CDK4(R24C) mice bearing multiple autochthonous melanomas did not reject transplanted B16 melanoma despite treatment with Ad-hTRP2 and TLR ligands, suggesting the development of tumor immunotolerance. Further investigations in our novel genetic melanoma model may help to better understand the role of the immune system in the pathogenesis and treatment of this life-threatening disease. PMID- 16707472 TI - DNA fusion vaccines induce epitope-specific cytotoxic CD8(+) T cells against human leukemia-associated minor histocompatibility antigens. AB - The graft-versus-leukemia effect of allogeneic stem-cell transplantation is believed to be mediated by T-cell recognition of minor histocompatibility antigens on recipient cells. For minor histocompatibility antigens HA-1 and HA-2, normal cell expression is restricted to hemopoietic cells, and boosting the immune response to these antigens may potentiate graft-versus-leukemia effect without accompanying graft-versus-host disease. To increase efficacy, expansion of HA-1- or HA-2-specific CTL before transplantation is desirable. However, primary HA-1- or HA-2-specific CTL expanded in vitro are often of low avidity. An alternative approach is to prime specific CTL responses in vivo by vaccination. Clearly, donor vaccination must be safe and specific. We have developed DNA fusion vaccines able to induce high levels of epitope-specific CTL using linked CD4(+) T-cell help. The vaccines incorporate a domain of tetanus toxin (DOM) fused to a sequence encoding a candidate MHC class I binding peptide. This design generates antitumor CD8(+) T-cell responses and protective immunity in preclinical models. For clinical application, we constructed vaccines encoding HLA-A*0201-restricted peptides from human HA-1 and HA-2, which were fused to DOM, and tested their performance in HLA-A*0201-transgenic mice. Priming induced epitope-specific, IFNgamma-producing CD8(+) T cells with cytotoxic function boosted to high levels with electroporation. Strikingly, these mouse T cells efficiently killed human lymphoblastoid cell lines expressing endogenous HA-1 or HA-2. High avidity is indicated by the independence of cytolysis from CD8/MHC class I interaction. These safe epitope-specific vaccines offer a potential strategy to prime HA-1- or HA-2-specific CTL in transplant donors before adoptive transfer. PMID- 16707473 TI - Intratumor CD4 T-cell accumulation requires stronger priming than for expansion and lymphokine secretion. AB - T cells need to migrate to and accumulate inside tumors before mediating rejection of the tumor. The number of specific T cells inside tumors may depend on the efficiency of priming in the draining lymph node (DLN), intratumor deletion, suppressive phenomena, or both. We used monoclonal anti-male antigen CD4 (Marilyn) T cells and tumor cell lines expressing or not the corresponding antigen (Dby) to analyze CD4 T-cell accumulation in tumors. Priming by MHC II(+) or MHC II(-) male splenocytes or Dby(+) tumor cells induced similar Marilyn T cell expansion in the DLN and recirculation in other lymph nodes and capacity to produce IFN-gamma. However, intratumor accumulation was different for each priming condition. In mice with Dby(-) tumors, MHC II(+) male splenocyte priming induced greater, although not statistically significant, Marilyn T-cell accumulation in the tumors than MHC II(-) male splenocyte priming. In mice with Dby(+) tumors, priming in the tumor DLN induced less Marilyn T-cell intratumor accumulation than priming by MHC II(+) male splenocytes. We saw comparable differences for Marilyn T-cell accumulation in gut lamina propria, suggesting that priming affects effector T-cell accumulation in inflamed tissues. Mature dendritic cells were loaded with graded doses of Dby peptide to control for antigen-presenting cell characteristics during priming. We observed similar proliferation, with higher concentrations inducing higher intratumor accumulation. Thus, intratumor accumulation requires stronger stimulation than for proliferation or the capacity to secrete lymphokines. In this system, priming intensity alone can explain the number of intratumor T cells without having to call for intratumor deletion or suppression phenomena. PMID- 16707474 TI - Vaccination with human HER-2/neu (435-443) CTL peptide induces effective antitumor immunity against HER-2/neu-expressing tumor cells in vivo. AB - HER-2/neu is a self-antigen expressed by tumors and nonmalignant epithelial tissues. The possibility of self-tolerance to HER-2/neu-derived epitopes has raised questions concerning their utility in antitumor immunotherapy. Altered HER 2/neu peptide ligands capable of eliciting enhanced immunity to tumor-associated HER-2/neu epitopes may circumvent this problem. The human CTL peptide HER-2/neu (435-443) [hHER-2(9(435))] represents a xenogeneic altered peptide ligand of its mouse homologue, differing by one amino acid residue at position 4. In contrast to mHER-2(9(435)), vaccination of HLA-A*0201 transgenic (HHD) mice with hHER 2(9(435)) significantly increased the frequency of mHER-2(9(435))-specific CTL and also induced strong protective and therapeutic immunity against the transplantable ALC tumor cell line transfected to coexpress HLA-A*0201 and hHER 2/neu or rHER-2/neu. Similar results were also obtained with wild-type C57BL/6 mice inoculated with HER-2/neu transfectants of ALC. Adoptive transfer of CD8(+) CTL from mice immunized with hHER-2(9(435)) efficiently protected naive syngeneic mice inoculated with ALC tumors. In conclusion, our results show that HER 2(9(435)) serves as a tumor rejection molecule. They also propose a novel approach for generating enhanced immunity against a self-HER-2/neu CTL epitope by vaccinating with xenogeneic altered peptide ligands and provide useful insights for the design of improved peptide-based vaccines for the treatment of patients with HER-2/neu-overexpressing tumors. PMID- 16707475 TI - Processing of tumor-associated antigen by the proteasomes of dendritic cells controls in vivo T-cell responses. AB - Dendritic cells are unique in their capacity to process antigens and prime naive CD8(+) T cells. Contrary to most cells, which express the standard proteasomes, dendritic cells express immunoproteasomes constitutively. The melanoma-associated protein Melan-A(MART1) contains an HLA-A2-restricted peptide that is poorly processed by melanoma cells expressing immunoproteasomes in vitro. Here, we show that the expression of Melan-A in dendritic cells fails to elicit T-cell responses in vitro and in vivo because it is not processed by the proteasomes of dendritic cells. In contrast, dendritic cells lacking immunoproteasomes induce strong anti-Melan-A T-cell responses in vitro and in vivo. These results suggest that the inefficient processing of self-antigens, such as Melan-A, by the immunoproteasomes of professional antigen-presenting cells prevents the induction of antitumor T-cell responses in vivo. PMID- 16707476 TI - Accelerated tumor formation in a fatless mouse with type 2 diabetes and inflammation. AB - Epidemiologic studies show a positive association between obesity and cancer risk. In addition to increased body adiposity and secretion of fat-derived hormones, obesity is also linked to insulin resistance, type 2 diabetes, and chronic inflammation. We used the fatless A-ZIP/F-1 transgenic mouse to dissociate the relative role of each of these underlying factors in the development of cancer. These mice are unique in that they do not have white fat but do develop type 2 diabetes. In two cancer models, the classic two-stage skin carcinogenesis protocol and the C3(1)/T-Ag transgenic mouse mammary tumor model, A-ZIP/F-1 mice displayed higher tumor incidence, tumor multiplicity, and decreased tumor latency than wild-type mice. We examined circulating levels of adipokines, growth factors, and cytokines. As expected, adipokines (i.e., leptin, adiponectin, and resistin) were undetectable or found at very low levels in the blood of fatless mice. However, insulin, insulin-like growth factor-I, growth hormone, vascular endothelial growth factor, and proinflammatory Th2 cytokines, such as interleukin (IL)-1beta, IL-4, and IL-6, were elevated in A-ZIP/F-1 mice. Additionally, we examined multiple phosphorylated proteins (i.e., protein kinase B/Akt and ErbB2/HER-2 kinase) associated with cancer development. Results show that many of these phosphorylated proteins were activated specifically in the A ZIP/F-1 skin but not in the wild-type skin. These findings suggest that adipokines are not required for the promotion of tumor development and thus contradict the epidemiologic data linking obesity to carcinogenesis. We postulate that insulin resistance and inflammation are responsible for the positive correlation with cancer observed in A-ZIP/F-1 mice. PMID- 16707477 TI - Therapeutic effect of imatinib in gastrointestinal stromal tumors: AKT signaling dependent and independent mechanisms. AB - Most gastrointestinal stromal tumors (GISTs) possess a gain-of-function mutation in c-KIT. Imatinib mesylate, a small-molecule inhibitor against several receptor tyrosine kinases, including KIT, platelet-derived growth factor receptor-alpha, and BCR-ABL, has therapeutic benefit for GISTs both via KIT and via unknown mechanisms. Clinical evidence suggests that a potential therapeutic benefit of imatinib might result from decreased glucose uptake as measured by positron emission tomography using 18-fluoro-2-deoxy-d-glucose. We sought to determine the mechanism of and correlation to altered metabolism and cell survival in response to imatinib. Glucose uptake, cell viability, and apoptosis in GIST cells were measured following imatinib treatment. Lentivirus constructs were used to stably express constitutively active AKT1 or AKT2 in GIST cells to study the role of AKT signaling in metabolism and cell survival. Immunoblots and immunofluorescent staining were used to determine the levels of plasma membrane-bound glucose transporter Glut4. We show that oncogenic activation of KIT maximizes glucose uptake in an AKT-dependent manner. Imatinib treatment markedly reduces glucose uptake via decreased levels of plasma membrane-bound Glut4 and induces apoptosis or growth arrest by inhibiting KIT activity. Importantly, expression of constitutively active AKT1 or AKT2 does not rescue cells from the imatinib mediated apoptosis although glucose uptake was not blocked, suggesting that the potential therapeutic effect of imatinib is independent of AKT activity and glucose deprivation. Overall, these findings contribute to a clearer understanding of the molecular mechanisms involved in the therapeutic benefit of imatinib in GIST and suggest that a drug-mediated decrease in tumor metabolism observed clinically may not entirely reflect therapeutic efficacy of treatment. PMID- 16707478 TI - Quantitative in situ analysis of beta-catenin expression in breast cancer shows decreased expression is associated with poor outcome. AB - The role of beta-catenin in breast cancer and its prognostic value is controversial. The prognostic value had been assessed previously in a series of nonquantitative immunohistochemical studies with conflicting results. In efforts to clarify the relationship between beta-catenin protein expression and breast cancer prognosis, we have assessed a retrospective 600 case cohort of breast cancer tumors from the Yale Pathology archives on tissue microarrays. They were assessed using automated quantitative analysis (AQUA) with a series of array embedded cell lines for which the beta-catenin concentration was standardized by an ELISA assay. The expression levels of the standard clinical markers HER2, estrogen receptor (ER), progesterone receptor (PR), and Ki-67 were also assessed on the same cohort. X-tile software was used to select optimal protein concentration cutpoints and to evaluate the outcome using a training set and a validation set. We found that low-level expression of membranous beta-catenin is associated with significantly worse outcome (38% versus 76%, 10-year survival, validation set log-rank P = 0.0016). Multivariate analysis of this marker, assessed in a proportional hazards model with tumor size, age, node status, nuclear grade, ER, PR, HER2, and Ki-67, is still highly significant with a hazard ratio of 6.8 (P < 0.0001, 95% confidence interval, 3.1-15.1). These results suggest that loss of beta-catenin expression at the membrane, as assessed by objective quantitative analysis methods, may be useful as a prognostic marker or may be part of a useful algorithm for prognosis in breast cancer. PMID- 16707479 TI - DNA methylation changes after 5-aza-2'-deoxycytidine therapy in patients with leukemia. AB - 5-Aza-2'-deoxycytidine (decitabine) is postulated to have clinical activity in myeloid leukemias via its ability to inhibit DNA methylation. To study this, we examined DNA methylation in patients with leukemia treated with decitabine. Five days after the treatment, total genomic 5-methylcytosine/cytosine decreased on average by 14% (from 4.3% to 3.7%), whereas methylation of repetitive DNA elements showed a mean decrease of 9% and 16% for Alu and long interspersed nucleotide elements, respectively. Methylation decreased linearly with increasing doses between 5 and 20 mg/m(2)/d (r = 0.88; P = 0.05) but showed a plateau above that. Hypomethylation correlated with response in patients with acute myelogenous leukemia treated with low doses (5-20 mg/m(2)/d), but patients with chronic myelogenous leukemia treated with high doses (100-180 mg/m(2)/d) showed no such correlation. Aberrant methylation of p15 (>10%) was found in 27% of patients, and 80% of these showed a decrease by at least one third, but this did not correlate with response. The imprinted gene H19 showed little change in methylation after decitabine. In conclusion, we show dose-dependent hypomethylation after decitabine at low doses. Increasing the dose, which has been shown previously to result in a reduced response rate, was not accompanied by further hypomethylation. PMID- 16707480 TI - HER-2/neu status is a determinant of mammary aromatase activity in vivo: evidence for a cyclooxygenase-2-dependent mechanism. AB - Cytochrome P450 aromatase (aromatase), a product of the CYP19 gene, catalyzes the synthesis of estrogens from androgens. Given the significance of estrogen synthesis in hormone-dependent breast carcinogenesis, it is important to elucidate the mechanisms that regulate CYP19 expression. The main objective of this study was to define the interrelationship between HER-2/neu, cyclooxygenase 2 (COX-2), and aromatase in mammary tissue. Mammary aromatase activity and prostaglandin E(2) (PGE(2)) levels were increased in mice with mammary-targeted expression of a COX-2 transgene. In vitro, overexpressing COX-2 caused both increased PGE(2) production and aromatase activity, effects that were suppressed by celecoxib, a selective COX-2 inhibitor. Previously, we found that overexpression of HER-2/neu was associated with increased levels of COX-2 in human breast cancers. Here, we show that overexpression of HER-2/neu is also associated with increased aromatase activity. These results suggested the possibility that COX-2 was the functional intermediate linking HER-2/neu and aromatase. Consistent with this idea, COX-2 deficiency led to a gene dose dependent reduction in mammary aromatase activity in a HER-2/neu transgenic mouse model. Complementary in vitro studies showed that HER-2/neu-mediated induction of PGE(2) synthesis and aromatase activity were suppressed by inhibiting COX-2. Collectively, our data indicate that COX-2 is the functional intermediate linking HER-2/neu and aromatase and suggest that inhibitors of PGE(2) synthesis will suppress estrogen biosynthesis in breast tissue. PMID- 16707481 TI - (-)-Epigallocatechin-3-gallate prevents photocarcinogenesis in mice through interleukin-12-dependent DNA repair. AB - We have shown previously that topical application of (-)-epigallocatechin-3 gallate (EGCG), the major polyphenol of green tea, prevents photocarcinogenesis in mice. EGCG prevents UVB-induced immunosuppression by inducing interleukin-12 (IL-12). As immunosuppression is a risk factor for photocarcinogenesis, we investigated the possibility that EGCG also prevents UVB-induced photocarcinogenesis through an IL-12-dependent DNA repair mechanism. To investigate this possibility, we determined the effects of EGCG on photocarcinogenesis in IL-12 knockout (KO) mice using the formation of cyclobutane pyrimidine dimers (CPD) as an indicator of the extent of UVB-induced DNA damage. Topical application of EGCG (1 mg/cm(2) skin) prevented photocarcinogenesis in wild-type (C3H/HeN) mice in terms of tumor incidence and tumor multiplicity but did not prevent photocarcinogenesis in IL-12 KO mice. UVB induced DNA damage, as determined by the formation of CPDs and the number of sunburn cells, was resolved more rapidly in the skin of wild-type mice treated with EGCG than untreated control mice. In contrast, the extent of UVB-induced DNA damage and the numbers of sunburn cells were not significantly different in the EGCG-treated IL-12 KO mice and untreated control mice. In addition, treatment of XPA-proficient human fibroblast cells with EGCG promoted repair of UVB-induced CPDs in a dose-dependent manner but not in an XPA-deficient cells, indicating that the nucleotide excision repair mechanism is involved in EGCG-mediated DNA repair. Taken together, these results indicate for the first time that EGCG can prevent photocarcinogenesis through an EGCG-induced IL-12-dependent DNA repair mechanism. PMID- 16707482 TI - A prospective study on habitual duration of sleep and incidence of breast cancer in a large cohort of women. AB - Mounting evidence suggests habitual sleep duration is associated with various health outcomes; both short and long sleep duration have been implicated in increased risk of cardiovascular disease, diabetes, and all-cause mortality. However, data on the relation between sleep duration and cancer risk are sparse and inconclusive. A link between low levels of melatonin, a hormone closely related to sleep, and increased risk of breast cancer has recently been suggested but it is unclear whether duration of sleep may affect breast cancer risk. We explored the association between habitual sleep duration reported in 1986 and subsequent risk of breast cancer in the Nurses' Health Study using Cox proportional hazards models. During 16 years of follow-up, 4,223 incident cases of breast cancer occurred among 77,418 women in this cohort. Compared with women sleeping 7 hours, covariate-adjusted hazard ratios and 95% confidence intervals for those sleeping < or =5, 6, 8, and > or =9 hours were 0.93 (0.79-1.09), 0.98 (0.91-1.06), 1.05 (0.97-1.13), and 0.95 (0.82-1.11), respectively. A moderate trend in risk increase towards longer sleep duration was observed when analyses were restricted to participants who reported same sleep duration in 1986 and 2000 (P(trend) = 0.05). In this prospective study, we found no convincing evidence for an association between sleep duration and the incidence of breast cancer. PMID- 16707483 TI - Analyzing proteomes and protein function using graphical comparative analysis of tandem mass spectrometry results. AB - Although generating large amounts of proteomic data using tandem mass spectrometry has become routine, there is currently no single set of comprehensive tools for the rigorous analysis of tandem mass spectrometry results given the large variety of possible experimental aims. Currently available applications are typically designed for displaying proteins and posttranslational modifications from the point of view of the mass spectrometrist and are not versatile enough to allow investigators to develop biological models of protein function, protein structure, or cell state. In addition, storage and dissemination of mass spectrometry-based proteomic data are problems facing the scientific community. To address these issues, we have developed a relational database model that efficiently stores and manages large amounts of tandem mass spectrometry results. We have developed an integrated suite of multifunctional analysis software for interpreting, comparing, and displaying these results. Our system, Bioinformatic Graphical Comparative Analysis Tools (BIGCAT), allows sophisticated analysis of tandem mass spectrometry results in a biologically intuitive format and provides a solution to many data storage and dissemination issues. PMID- 16707484 TI - cAMP-response element-binding protein mediates acid-induced NADPH oxidase NOX5-S expression in Barrett esophageal adenocarcinoma cells. AB - Gastroesophageal reflux disease complicated by Barrett esophagus (BE) is a major risk factor for esophageal adenocarcinoma (EA). The mechanisms whereby acid reflux may accelerate the progression from BE to EA are not known. We found that NOX1 and NOX5-S were the major isoforms of NADPH oxidase in SEG1-EA cells. The expression of NOX5-S mRNA was significantly higher in these cells than in esophageal squamous epithelial cells. NOX5 mRNA was also significantly higher in Barrett tissues with high grade dysplasia than without dysplasia. Pulsed acid treatment significantly increased H(2)O(2) production in both SEG1-EA cells and BE mucosa, which was blocked by the NADPH oxidase inhibitor apocynin. In SEG1 cells, acid treatment increased mRNA expression of NOX5-S, but not NOX1, and knockdown of NOX5 by NOX5 small interfering RNA abolished acid-induced H(2)O(2) production. In addition, acid treatment increased intracellular Ca(2+) and phosphorylation of cAMP-response element-binding protein (CREB). Acid-induced NOX5-S expression and H(2)O(2) production were significantly inhibited by removal of extracellular Ca(2+) and by knockdown of CREB using CREB small interfering RNA. Two novel CREB-binding elements TGACGAGA and TGACGCTG were identified in the NOX5-S gene promoter. Overexpression of CREB significantly increased NOX5-S promoter activity. Knockdown of NOX5 significantly decreased [(3)H]thymidine incorporation, which was restored by 10(-13) M H(2)O(2). Knockdown of NOX5 also significantly decreased retinoblastoma protein phosphorylation and increased cell apoptosis and caspase-9 expression. In conclusion, in SEG1 EA cells NOX5-S is overexpressed and mediates acid-induced H(2)O(2) production. Acid-induced NOX5-S expression depends on an increase in intracellular Ca(2+) and activation of CREB. NOX5-S contributes to increased cell proliferation and decreased apoptosis. PMID- 16707485 TI - Individual Cas phosphorylation sites are dispensable for processive phosphorylation by Src and anchorage-independent cell growth. AB - Cas is a multidomain signaling protein that resides in focal adhesions. Cas possesses a large central substrate domain containing 15 repeats of the sequence YXXP, which are phosphorylated by Src. The phosphorylation sites are essential for the roles of Cas in cell migration and in regulation of the actin cytoskeleton. We showed previously that Src catalyzes the multisite phosphorylation of Cas via a processive mechanism. In this study, we created mutant forms of Cas to identify the determinants for processive phosphorylation. Mutants containing single or multiple YXXP mutations were phosphorylated processively by Src, suggesting that individual sites are dispensable. The results also suggest that there is no defined order to the Cas phosphorylation events. We also studied the effects of these mutations by reintroducing Cas into Cas-deficient fibroblasts. Mutants lacking some or all YXXP sites augment the ability of Src to promote anchorage-independent growth. On the other hand, deletion of YXXP sites compromises the ability of Cas to promote tumor cell migration. PMID- 16707486 TI - Pigment epithelium-derived factor inhibits advanced glycation end product-induced retinal vascular hyperpermeability by blocking reactive oxygen species-mediated vascular endothelial growth factor expression. AB - Pigment epithelium-derived factor (PEDF) is the most potent inhibitor of angiogenesis, suggesting that loss of PEDF contributes to proliferative diabetic retinopathy. However, the role of PEDF against retinal vascular hyperpermeability remains to be elucidated. We investigated here whether and how PEDF could inhibit the advanced glycation end product (AGE) signaling to vascular hyperpermeability. Intravenous administration of AGEs to normal rats not only increased retinal vascular permeability by stimulating vascular endothelial growth factor (VEGF) expression but also decreased retinal PEDF levels. Simultaneous treatments with PEDF inhibited the AGE-elicited VEGF-mediated permeability by down-regulating mRNA levels of p22(phox) and gp91(phox), membrane components of NADPH oxidase, and subsequently decreasing retinal levels of an oxidative stress marker, 8 hydroxydeoxyguanosine. PEDF also inhibited the AGE-induced vascular hyperpermeability evaluated by transendothelial electrical resistance by suppressing VEGF expression. Furthermore, PEDF decreased reactive oxygen species (ROS) generation in AGE-exposed endothelial cells by suppressing NADPH oxidase activity via down-regulation of mRNA levels of p22(PHOX) and gp91(PHOX). This led to blockade of the AGE-elicited Ras activation and NF-kappaB-dependent VEGF gene induction in endothelial cells. These results indicate that the central mechanism for PEDF inhibition of the AGE signaling to vascular permeability is by suppression of NADPH oxidase-mediated ROS generation and subsequent VEGF expression. Substitution of PEDF may offer a promising strategy for halting the development of diabetic retinopathy. PMID- 16707487 TI - H2A.Z stabilizes chromatin in a way that is dependent on core histone acetylation. AB - The functional and structural chromatin roles of H2A.Z are still controversial. This work represents a further attempt to resolve the current functional and structural dichotomy by characterizing chromatin structures containing native H2A.Z. We have analyzed the role of this variant in mediating the stability of the histone octamer in solution using gel-filtration chromatography at different pH. It was found that decreasing the pH from neutral to acidic conditions destabilized the histone complex. Furthermore, it was shown that the H2A.Z-H2B dimer had a reduced stability. Sedimentation velocity analysis of nucleosome core particles (NCPs) reconstituted from native H2A.Z-containing octamers indicated that these particles exhibit a very similar behavior to that of native NCPs consisting of canonical H2A. Sucrose gradient fractionation of native NCPs under different ionic strengths indicated that H2A.Z had a subtle tendency to fractionate with more stabilized populations. An extensive analysis of the salt dependent dissociation of histones from hydroxyapatite-adsorbed chromatin revealed that, whereas H2A.Z co-elutes with H3-H4, hyperacetylation of histones (by treatment of chicken MSB cells with sodium butyrate) resulted in a significant fraction of this variant eluting with the canonical H2A. These studies also showed that the late elution of this variant (correlated to enhanced binding stability) was independent of the chromatin size and of the presence or absence of linker histones. PMID- 16707488 TI - Type Igamma661 phosphatidylinositol phosphate kinase directly interacts with AP2 and regulates endocytosis. AB - Clathrin-coated vesicles mediate sorting and intracellular transport of membrane bound proteins. The formation of these coats is initiated by the assembly of adaptor proteins (AP), which specifically bind to membrane cargo proteins via recognition of endocytic sorting motifs. The lipid signaling molecule phosphatidylinositol 4,5-bisphosphate (PI(4,5)P(2)) is critical for this process, as it serves as both a targeting and regulatory factor. PI(4,5)P(2) is synthesized by type I phosphatidylinositol phosphate kinases (PIPKI). We have discovered a direct interaction between the mu2-subunit of the AP2 complex and PIPKIgamma661 via a yeast two-hybrid screen. This interaction was confirmed using both the mu2-subunit in glutathione S-transferase pulldowns and via coimmunoprecipitation of endogenous PIPKIgamma661 with the AP2 complex from HEK293 cells. The interaction is mediated, in vivo, by a tyrosine-based motif in the 26-amino acid tail of PIPKIgamma661. Because AP2 regulates endocytosis of transferrin receptor from the plasma membrane, we also examined a role for PIPKIgamma661 using a flow cytometry endocytosis assay. We observed that stable expression of wild type PIPKIgamma661 in Madin-Darby canine kidney cells enhanced transferrin uptake, whereas stable expression of kinase-dead PIPKIgamma661 had an inhibitory effect. Neither condition affected the overall cellular level of PI(4,5)P(2). RNA interference-based knockdown of PIPKIgamma661 in HeLa cells also had an inhibitory effect on transferrin endocytosis using the same assay system. Collectively, this evidence implies an important role for PIPKIgamma661 in the AP2-mediated endocytosis of transferrin. PMID- 16707489 TI - Catalytic mechanism and structure of viral flavin-dependent thymidylate synthase ThyX. AB - By using biochemical and structural analyses, we have investigated the catalytic mechanism of the recently discovered flavin-dependent thymidylate synthase ThyX from Paramecium bursaria chlorella virus-1 (PBCV-1). Site-directed mutagenesis experiments have identified several residues implicated in either NADPH oxidation or deprotonation activity of PBCV-1 ThyX. Chemical modification by diethyl pyrocarbonate and mass spectroscopic analyses identified a histidine residue (His53) crucial for NADPH oxidation and located in the vicinity of the redox active N-5 atom of the FAD ring system. Moreover, we observed that the conformation of active site key residues of PBCV-1 ThyX differs from earlier reported ThyX structures, suggesting structural changes during catalysis. Steady state kinetic analyses support a reaction mechanism where ThyX catalysis proceeds via formation of distinct ternary complexes without formation of a methyl enzyme intermediate. PMID- 16707490 TI - A conserved 12-amino acid motif in Sall1 recruits the nucleosome remodeling and deacetylase corepressor complex. AB - Sall1 is a multi-zinc finger transcription factor that represses gene expression and regulates organogenesis. In this report, we further characterize the domain of Sall1 necessary for repression. We show that endogenous Sall1 binds to the nucleosome remodeling and deacetylase corepressor complex (NuRD) and confirm the functionality of the Sall1-associating macromolecular complex by showing that the complex possesses HDAC activity. NuRD is involved in global transcriptional repression and regulation of specific developmental processes. The mechanism by which sequence-specific DNA-binding proteins associate with NuRD is not well understood. We have identified a highly conserved 12-amino acid motif in the transcription factor Sall1 that is sufficient for the recruitment of NuRD. Single amino acid substitutions defined the critical amino acid peptide motif as RRKQXK PXXF. This motif probably exhibits a more general role in regulating gene expression, since other proteins containing this domain, including all Sall family members and an unrelated zinc finger protein Ebfaz, mediate transcriptional repression and associate with NuRD. These results also have important implications for the pathogenesis of Townes-Brocks, a syndrome caused by SALL1 mutations. PMID- 16707491 TI - Concerted action of Smad and CREB-binding protein regulates bone morphogenetic protein-2-stimulated osteoblastic colony-stimulating factor-1 expression. AB - Bone remodeling depends upon proper osteoblast and osteoclast function. Bone morphogenetic protein-2 (BMP-2) stimulates differentiation of osteoblasts from pluripotent precursors. Osteoclast formation depends on the concerted action of osteoblast-derived receptor activator of NF-kappaB ligand and colony-stimulating factor-1 (CSF-1). BMP-2 stimulates receptor activator of NF-kappaB ligand expression. However, the effect of BMP-2 on CSF-1 expression has not been studied. We investigated the role of BMP-2 in CSF-1 expression in osteogenic C2C12 cells. Incubation of C2C12 cells with BMP-2 supported osteoclastogenesis of spleen cells with a concomitant increase in expression of CSF-1 mRNA and protein. To determine the mechanism, we identified a BMP-responsive element between -627 bp and -509 bp in the CSF-1 promoter. DNase I footprint analysis revealed the presence of consensus Smad binding motif in this region. Electrophoretic mobility shift assay showed BMP-2-stimulated binding of proteins to this motif. Mutation of core sequence as well as its 5'- and 3'-flanking sequences abolished the DNA protein interaction resulting in inhibition of CSF-1 transcription. Supershift analysis detects the presence of Smads 1, 5, and 4 and the transcriptional coactivator CREB-binding protein in the BMP-responsive element-protein complex. In addition, Smads 1 and 5 alone or in combination with Smad 4 increased CSF-1 transcription. Furthermore, CREB-binding protein markedly increased transcription of CSF-1. These data represent the first evidence that BMP-2 increases the osteoclastogenic CSF-1 expression by a transcriptional mechanism using the canonical Smad pathway and provide a mechanism for BMP-2-induced osteoclast differentiation. PMID- 16707492 TI - Altering biomineralization by protein design. AB - To create a bioceramic with unique materials properties, biomineralization exploits cells to create a tissue-specific protein matrix to control the crystal habit, timing, and position of the mineral phase. The biomineralized covering of vertebrate teeth is enamel, a distinctive tissue of ectodermal origin that is collagen-free. In forming enamel, amelogenin is the abundant protein that undergoes self-assembly to contribute to a matrix that guides its own replacement by mineral. Conserved domains in amelogenin suggest their importance to biomineralization. We used gene targeting in mice to replace native amelogenin with one of two engineered amelogenins. Replacement changed enamel organization by altering protein-to-crystallite interactions and crystallite stacking while diminishing the ability of the ameloblast to interact with the matrix. These data demonstrate that ameloblasts must continuously interact with the developing matrix to provide amelogenin-specific protein to protein, protein to mineral, and protein to membrane interactions critical to biomineralization and enamel architecture while suggesting that mutations within conserved amelogenin domains could account for enamel variations preserved in the fossil record. PMID- 16707493 TI - Slug regulates integrin expression and cell proliferation in human epidermal keratinocytes. AB - The human epidermis is a self-renewing epithelial tissue composed of several layers of keratinocytes. Within the epidermis there exists a complex array of cell adhesion structures, and many of the cellular events within the epidermis (differentiation, proliferation, and migration) require that these adhesion structures be remodeled. The link between cell adhesion, proliferation, and differentiation within the epidermis is well established, and in particular, there is strong evidence to link the process of terminal differentiation to integrin adhesion molecule expression and function. In this paper, we have analyzed the role of a transcriptional repressor called Slug in the regulation of adhesion molecule expression and function in epidermal keratinocytes. We report that activation of Slug, which is expressed predominantly in the basal layer of the epidermis, results in down-regulation of a number of cell adhesion molecules, including E-cadherin, and several integrins, including alpha3, beta1, and beta4. We demonstrate that Slug binds to the alpha3 promoter and that repression of alpha3 transcription by Slug is dependent on an E-box sequence within the promoter. This reduction in integrin expression is reflected in decreased cell adhesion to fibronectin and laminin-5. Despite the reduction in integrin expression and function, we do not observe any increase in differentiation. We do, however, find that activation of Slug results in a significant reduction in keratinocyte proliferation. PMID- 16707494 TI - Specific recognition of apoptotic cells reveals a ubiquitous and unconventional innate immunity. AB - The purpose of physiological cell death is the noninflammatory clearance of cells that have become inappropriate or nonfunctional. Consistent with this function, the recognition of apoptotic cells by professional phagocytes, including macrophages and dendritic cells, triggers a set of potent anti-inflammatory responses manifest on multiple levels. The immediate-early inhibition of proinflammatory cytokine gene transcription in the phagocyte is a proximate consequence of recognition of the apoptotic corpse, independent of subsequent engulfment and soluble factor involvement. Here, we show that recognition is linked to a characteristic signature of responses, including MAPK signaling events and the ablation of proinflammatory transcription and cytokine secretion. Specific recognition and response occurs without regard to the origin (species, tissue type, or suicidal stimulus) of the apoptotic cell and does not involve Toll-like receptor signaling. These features mark this as an innate immunity fundamentally distinct from the discrimination of "self" versus "other" considered to be the hallmark of conventional immunity. This profound unconventional innate immune discrimination of effete from live cells is as ubiquitous as apoptotic cell death itself, manifest by professional and nonprofessional phagocytes and nonphagocytic cell types alike. Innate apoptotic immunity provides an intrinsic anti-inflammatory circuit that attenuates proinflammatory responses dynamically and may act systemically as a powerful physiological regulator of immunity. PMID- 16707495 TI - Metabolic changes in glucose transporter-deficient Leishmania mexicana and parasite virulence. AB - Leishmania mexicana are parasitic protozoa that express a variety of glycoconjugates that play important roles in their biology as well as the storage carbohydrate beta-mannan, which is an essential virulence factor for survival of intracellular amastigote forms in the mammalian host. Glucose transporter null mutants, which are viable as insect form promastigotes but not as amastigotes, do not take up glucose and other hexoses but are still able to synthesize these glycoconjugates and beta-mannan, although at reduced levels. Synthesis of these carbohydrate-containing macromolecules could be accounted for by incorporation of non-carbohydrate precursors into carbohydrates by gluconeogenesis. However, the significantly reduced level of the virulence factor beta-mannan in the glucose transporter null mutants compared with wild-type parasites may contribute to the non-viability of these null mutants in the disease-causing amastigote stage of the life cycle. PMID- 16707496 TI - The UBA domains of NUB1L are required for binding but not for accelerated degradation of the ubiquitin-like modifier FAT10. AB - Proteins selected for degradation are labeled with multiple molecules of ubiquitin and are subsequently cleaved by the 26 S proteasome. A family of proteins containing at least one ubiquitin-associated (UBA) domain and one ubiquitin-like (UBL) domain have been shown to act as soluble ubiquitin receptors of the 26 S proteasome and introduce a new level of specificity into the degradation system. They bind ubiquitylated proteins via their UBA domains and the 26 S proteasome via their UBL domain and facilitate the contact between substrate and protease. NEDD8 ultimate buster-1 long (NUB1L) belongs to this class of proteins and contains one UBL and three UBA domains. We recently reported that NUB1L interacts with the ubiquitin-like modifier FAT10 and accelerates its degradation and that of its conjugates. Here we show that a deletion mutant of NUB1L lacking the UBL domain is still able to bind FAT10 but not the proteasome and no longer accelerates FAT10 degradation. A version of NUB1L lacking all three UBA domains, on the other hand, looses the ability to bind FAT10 but is still able to interact with the proteasome and accelerates the degradation of FAT10. The degradation of a FAT10 mutant containing only the C terminal UBL domain is also still accelerated by NUB1L, even though the two proteins do not interact. In addition, we show that FAT10 and either one of its UBL domains alone can interact directly with the 26 S proteasome. We propose that NUB1L not only acts as a linker between the 26 S proteasome and ubiquitin-like proteins, but also as a facilitator of proteasomal degradation. PMID- 16707497 TI - The role of the phospho-CDK2/cyclin A recruitment site in substrate recognition. AB - Phospho-CDK2/cyclin A, a kinase that is active in cell cycle S phase, contains an RXL substrate recognition site that is over 40 A from the catalytic site. The role of this recruitment site, which enhances substrate affinity and catalytic efficiency, has been investigated using peptides derived from the natural substrates, namely CDC6 and p107, and a bispeptide inhibitor in which the gamma phosphate of ATP is covalently attached by a linker to the CDC6 substrate peptide. X-ray studies with a 30-residue CDC6 peptide in complex with pCDK2/cyclin A showed binding of a dodecamer peptide at the recruitment site and a heptapeptide at the catalytic site, but no density for the linking 11 residues. Kinetic studies established that the CDC6 peptide had an 18-fold lower Km compared with heptapeptide substrate and that this effect required the recruitment peptide to be covalently linked to the substrate peptide. X-ray studies with the CDC6 bispeptide showed binding of the dodecamer at the recruitment site and the modified ATP in two alternative conformations at the catalytic site. The CDC6 bispeptide was a potent inhibitor competitive with both ATP and peptide substrate of pCDK2/cyclin A activity against a heptapeptide substrate (Ki = 0.83 nm) but less effective against RXL-containing substrates. We discuss how localization at the recruitment site (KD 0.4 microm) leads to increased catalytic efficiency and the design of a potent inhibitor. The notion of a flexible linker between the sites, which must have more than a minimal number of residues, provides an explanation for recognition and discrimination against different substrates. PMID- 16707498 TI - Absence of erythroblast macrophage protein (Emp) leads to failure of erythroblast nuclear extrusion. AB - In mammals, the functional unit for definitive erythropoiesis is the erythroblastic island, a multicellular structure composed of a central macrophage surrounded by developing erythroblasts. Erythroblast-macrophage interactions play a central role in the terminal maturation of erythroblasts, including enucleation. One possible mediator of this cell-cell interaction is the protein Emp (erythroblast macrophage protein). We used targeted gene inactivation to define the function of Emp during hematopoiesis. Emp null embryos die perinatally and show profound alterations in the hematopoietic system. A dramatic increase in the number of nucleated, immature erythrocytes is seen in the peripheral blood of Emp null fetuses. In the fetal liver virtually no erythroblastic islands are observed, and the number of F4/80-positive macrophages is substantially reduced. Those present lack cytoplasmic projections and are unable to interact with erythroblasts. Interestingly, wild type macrophages can bind Emp-deficient erythroblasts, but these erythroblasts do not extrude their nuclei, suggesting that Emp impacts enucleation in a cell autonomous fashion. Previous studies have implicated the actin cytoskeleton and its reorganization in both erythroblast enucleation as well as in macrophage development. We demonstrate that Emp associates with F-actin and that this interaction is important in the normal distribution of F-actin in both erythroblasts and macrophages. Thus, Emp appears to be required for erythroblast enucleation and in the development of the mature macrophages. The availability of an Emp null model provides a unique experimental system to study the enucleation process and to evaluate the function of macrophages in definitive erythropoiesis. PMID- 16707499 TI - Poxvirus mRNA cap methyltransferase. Bypass of the requirement for the stimulatory subunit by mutations in the catalytic subunit and evidence for intersubunit allostery. AB - The guanine-N7 methyltransferase domain of vaccinia virus mRNA capping enzyme is a heterodimer composed of a catalytic subunit vD1-(540-844) and a stimulatory subunit vD12. The poxvirus enzyme can function in vivo in Saccharomyces cerevisiae in lieu of the essential cellular cap methyltransferase Abd1. Coexpression of both poxvirus subunits is required to complement the growth of abd1delta cells. We performed a genetic screen for mutations in the catalytic subunit that bypassed the requirement for the stimulatory subunit in vivo. We thereby identified missense changes in vicinal residues Tyr-752 (to Ser, Cys, or His) and Asn-753 (to Ile), which are located in the cap guanine-binding pocket. Biochemical experiments illuminated a mechanism of intersubunit allostery, whereby the vD12 subunit enhances the affinity of the catalytic subunit for AdoMet and the cap guanine methyl acceptor by 6- and 14-fold, respectively, and increases kcat by a factor of 4. The bypass mutations elicited gains of function in both vD12-independent and vD12-dependent catalysis of cap methylation in vitro when compared with wild-type vD1-(540-844). These results highlight the power of yeast as a surrogate model for the genetic analysis of interacting poxvirus proteins and demonstrate that the activity of an RNA processing enzyme can be augmented through selection and protein engineering. PMID- 16707500 TI - Interferon regulatory factor 8 regulates RANTES gene transcription in cooperation with interferon regulatory factor-1, NF-kappaB, and PU.1. AB - Interferon regulatory factor (IRF)-8 is a member of the IRF family of transcription factors important in interferon-gamma-mediated signaling and in the development and function of dendritic cells. Regulated on activation, normal T cell expressed and secreted (RANTES, or CCL5) is a member of the CC chemokine family of proteins, strongly chemoattractant for several important immune cell types in host defense against infectious agents and cancer. Here we report that RANTES expression in IRF-8-null macrophages stimulated with interferon-gamma and lipopolysaccharide is markedly decreased. IRF-8 can activate RANTES gene transcription in synergism with IRF-1. Interestingly, IRF-8 can activate RANTES transcription independently of IRF-1 through direct physical interactions with NF kappaB c-Rel and PU.1 via the NF-kappaB element located at -88 to -79 in vitro and in vivo. This study uncovers a novel role of IRF-8 in the regulation of RANTES gene expression and the underlying molecular mechanisms whereby IRF-8 interacts with several other important transcription factors to initiate innate immune responses to pathogenic and inflammatory challenges by activating the RANTES gene. PMID- 16707501 TI - A novel role for protein phosphatase 2A in receptor-mediated regulation of the cardiac sarcolemmal Na+/H+ exchanger NHE1. AB - G(q) protein-coupled receptor stimulation increases sarcolemmal Na(+)/H(+) exchanger (NHE1) activity in cardiac myocytes by an ERK/RSK-dependent mechanism, most likely via RSK-mediated phosphorylation of the NHE1 regulatory domain. Adenosine A(1) receptor stimulation inhibits this response through a G(i) protein mediated pathway, but the distal inhibitory signaling mechanisms are unknown. In cultured adult rat ventricular myocytes (ARVM), the A(1) receptor agonist cyclopentyladenosine (CPA) inhibited the increase in NHE1 phosphorylation induced by the alpha(1)-adrenoreceptor agonist phenylephrine, without affecting activation of the ERK/RSK pathway. CPA also induced significant accumulation of the catalytic subunit of type 2A protein phosphatase (PP2A(c)) in the particulate fraction, which contained the cellular NHE1 complement; this effect was abolished by pretreatment with pertussis toxin to inactivate G(i) proteins. Confocal immunofluorescence microscopic imaging of CPA-treated ARVM revealed significant co-localization of PP2A(c) and NHE1, in intercalated disc regions. In an in vitro assay, purified PP2A(c) dephosphorylated a GST-NHE1 fusion protein containing aa 625-747 of the NHE1 regulatory domain, which had been pre-phosphorylated by recombinant RSK; such dephosphorylation was inhibited by the PP2A-selective phosphatase inhibitor endothall. In intact ARVM, the ability of CPA to attenuate the phenylephrine-induced increase in NHE1 phosphorylation and activity was lost in the presence of endothall. These studies reveal a novel role for the PP2A holoenzyme in adenosine A(1) receptor-mediated regulation of NHE1 activity in ARVM, the mechanism of which appears to involve G(i) protein-mediated translocation of PP2A(c) and NHE1 dephosphorylation. PMID- 16707502 TI - Mechanical stretch modulates the promoter activity of the profibrotic factor CCN2 through increased actin polymerization and NF-kappaB activation. AB - The connective tissue growth factor known as CCN2 is an inducible, profibrotic molecule that becomes aberrantly expressed in mechanical overload-bearing tissues. In this study, we found that CCN2 gene expression is rapidly induced in cyclically stretched bladder smooth muscle cells (SMCs) in vitro and in the detrusor muscle of a mechanically overloaded bladder in a rat model of experimental urethral obstruction. The activity of CCN2 promoter constructs, transiently transfected into cultured SMCs, was increased (up to 6-fold) by continuous cyclic stretching. Molecular analyses of the CCN2 promoter by serial construct deletions, cis-element mutagenesis, and electrophoretic mobility shift assays revealed that a highly conserved NF-kappaB binding site located within the CCN2 proximal promoter region is responsible for the activation of the promoter by stretch. Chromatin immunoprecipitation assays showed that NF-kappaB binds to the endogenous CCN2 promoter in both stretched cells and mechanically overloaded bladder tissues. Furthermore, stretch-dependent CCN2 promoter activity was significantly reduced upon inhibition of either phosphatidylinositol 3-kinase, p38 stress-activated kinase, or RhoA GTPase and was completely abolished upon inhibition of actin polymerization. Concordantly, actin polymerization was increased in either mechanically stretched cells or overloaded bladder tissues. Incubation of cultured SMCs with a cell-penetrating peptide containing the N terminal sequence, Ac-EEED, of smooth muscle alpha-actin, altered both actin cytoskeleton organization and stretch-mediated nuclear relocation of NF-kappaB, and subsequently, it reduced CCN2 promoter activity. Thus, mechanical stretch induced changes in actin dynamics mediate NF-kappaB activation and induce CCN2 gene expression, which probably initiates the fibrotic reactions observed in mechanical overload-associated pathologies. PMID- 16707503 TI - Identification of a novel inhibitory actin-capping protein binding motif in CD2 associated protein. AB - CD2-associated protein (CD2AP) is a scaffold molecule that plays a critical role in the maintenance of the kidney filtration barrier. Little, however, is understood about its mechanism of function. We used mass spectrometry to identify CD2AP-interacting proteins. Many of the proteins that we identified suggest a role for CD2AP in endocytosis and actin regulation. To address the role of CD2AP in regulation of the actin cytoskeleton, we focused on characterizing the interaction of CD2AP with actin-capping protein CP. We identified a novel binding motif LXHXTXXRPK(X)6P present in CD2AP that is also found in its homolog Cin85 and other capping protein-associated proteins such as CARMIL and CKIP-1. CD2AP inhibits the function of capping protein in vitro. Therefore, our results support a role of CD2AP in the regulation of the actin cytoskeleton. PMID- 16707504 TI - Transforming growth factor beta regulates the expression of the M2 muscarinic receptor in atrial myocytes via an effect on RhoA and p190RhoGAP. AB - Transforming growth factor beta (TGFbeta) signaling is involved in the development and regulation of multiple organ systems and cellular signaling pathways. We recently demonstrated that TGFbeta regulates the response of atrial myocytes to parasympathetic stimulation. Here, TGFbeta(1) is shown to inhibit expression of the M(2) muscarinic receptor (M(2)), which plays a critical role in the parasympathetic response of the heart. This effect is mimicked by overexpression of a dominant negative mutant of RhoA and by the RhoA kinase inhibitor Y27632, whereas adenoviral expression of a dominant activating-RhoA reverses TGFbeta inhibition of M(2) expression. TGFbeta(1) also mediates a decrease in GTP-bound RhoA and a reciprocal increase in the expression of the RhoA GTPase-activating protein, p190RhoGAP, whereas total RhoA is unchanged. Inhibition of M(2) promoter activity by TGFbeta(1) is mimicked by overexpression of p190RhoGAP, whereas a dominant negative mutant of p190RhoGAP reverses this effect of TGFbeta(1). In contrast to atrial myocytes, in mink lung epithelial cells, in which TGFbeta signaling through activation of RhoA has been previously identified, TGFbeta(1) stimulated an increase in GTP-bound RhoA in association with a reciprocal decrease in the expression of p190RhoGAP. Both effects demonstrated a similar dose dependence on TGFbeta(1). Thus TGFbeta regulation of M(2) muscarinic receptor expression is dependent on RhoA, and TGFbeta regulation of p190RhoGAP expression may be a cell type-specific mechanism for TGFbeta signaling through RhoA. PMID- 16707505 TI - Changes in lipid status and glass properties in cotyledons of developing sunflower seeds. AB - Biochemical events involved in the acquisition of germinability and storability during orthodox seed development are well documented; however, the roles played by the physical organization of lipids and water are poorly characterized. The aim of this work was to determine, using a thermodynamic approach, whether changes in thermal properties of lipid reserves, and intracellular glasses might play a role in sunflower (Helianthus annuus L.) seed development. Triacyglycerols (TAGs) accumulated in cotyledons until the end of seed filling, which occurred 42 days after anthesis (DAA). Further seed development, leading to mature seed at 58 DAA, was mainly associated with an enlargement of lipid bodies without significant changes either in the lipid content or in their composition. When cooled to -100 degrees C, lipid reserves from cotyledons of mature seeds displayed alpha and beta' polymorphic crystalline structures; however, the ability to form alpha crystals, which was an indicator of lipid purity, progressively appeared during seed development. Characteristics of lipid melting confirmed that seed maturation drying was associated with changes in TAG physical organization. Cotyledon development was associated with an increase in the temperature of glass to rubber transition (Tg), thus suggesting a decrease in molecular mobility during maturation drying. This phenomenon was concomitant with an increase in raffinose content. Our results demonstrate that physical characteristics of lipid reserves and glasses of sunflower cotyledons are developmentally regulated and might play a role in acquisition of seed germinability and storability. PMID- 16707506 TI - Predictive factors of trophoblastic invasion into the ampullary region of the tubal wall in ectopic pregnancy. AB - BACKGROUND: Predictive factors of damage to the Fallopian tube may guide the treatment for patients with tubal pregnancy. The aim of this study was to assess the predictive value of the following parameters: gestational age, serum beta-hCG concentrations and ultrasound findings. METHODS: A total of 105 patients with ampullary pregnancy undergoing salpingectomy were analysed. Trophoblastic invasion was histologically classified as stage I when limited to the tubal mucosa, stage II when extending to the muscle layer and stage III in the case of complete tubal wall infiltration. We correlated the depth of trophoblastic infiltration into the tubal wall with gestational age, beta-hCG concentration on the day of surgery and the type and size of the ectopic mass upon ultrasound. RESULTS: No association was observed between the depth of trophoblastic invasion and gestational age (P = 0.53) or tubal mass diameter (P = 0.43). Trophoblastic invasion was, however, associated with beta-hCG concentration (P < 0.001) and with the type of ultrasonographic image (P = 0.001). Beta-hCG levels of 2400 mIU/ml showed 82.8% sensitivity and 85.5% specificity for stage I, and levels of 5990 mIU/ml showed 82.6% sensitivity and 74.6% specificity for stage III. CONCLUSIONS: Depth of trophoblastic penetration into the tubal wall of the ampullary region of Fallopian tube is correlated with beta-hCG concentration and the type of ultrasonographic image; serum beta-hCG is the best predictor of the depth of penetration. PMID- 16707507 TI - Evidence-based guidelines for the investigation and medical treatment of recurrent miscarriage. AB - Recurrent miscarriage (RM; > or =3 consecutive early pregnancy losses) affects around 1% of fertile couples. Parental chromosomal anomalies, maternal thrombophilic disorders and structural uterine anomalies have been directly associated with recurrent miscarriage; however, in the vast majority of cases the pathophysiology remains unknown. We have updated the ESHRE Special Interest Group for Early Pregnancy (SIGEP) protocol for the investigation and medical management of RM. Based on the data of recently published large randomized controlled trials (RCTs) and meta-analyses, we recommend that basic investigations of a couple presenting with recurrent miscarriage should include obstetric and family history, age, BMI and exposure to toxins, full blood count, antiphospholipid antibodies (lupus anticoagulant and anticardiolipin antibodies), parental karyotype, pelvic ultrasound and/or hysterosalpingogram. Other investigations should be limited to particular cases and/or used within research programmes. Tender loving care and health advice are the only interventions that do not require more RCTs. All other proposed therapies, which require more investigations, are of no proven benefit or are associated with more harm than good. PMID- 16707508 TI - Effect of patient completed agenda forms and doctors' education about the agenda on the outcome of consultations: randomised controlled trial. AB - OBJECTIVE: To assess the effect of patient completed agenda forms for the consultation and doctors' education on identifying patients' agendas on the outcome of consultations. DESIGN: Randomised controlled trial. SETTING: General practices in Leicestershire and Nottinghamshire, United Kingdom. PARTICIPANTS: 46 general practitioners and 976 patients. INTERVENTIONS: Education for general practitioners, with an embedded clustered randomised controlled trial of a patient agenda form. MAIN OUTCOME MEASURES: Number of problems identified, time required to manage each problem, duration of consultations, number of problems raised after the doctor considered the consultation finished ("by the way" questions), and patient satisfaction. RESULTS: Data were available from 45 doctors (98%) and 857 patients (88%). The number of problems identified in each consultation increased by 0.2 (95% confidence interval 0.1 to 0.4) with the agenda form, by 0.3 (0.1 to 0.6) with education, and by 0.5 (0.3 to 0.7) with both interventions. The time required to manage each problem was not affected. The duration of consultations with the agenda form was increased by 0.9 minutes (0.3 to 1.5 minutes) and with the combined intervention by 1.9 minutes (1.0 to 2.8 minutes). Patient satisfaction with the depth of the doctor-patient relationship was increased with the agenda form. The occurrence of "by the way" presentations did not change. CONCLUSION: A patient completed agenda form before the consultation or general practitioner education about the agenda form, or both, enabled the identification of more problems in consultations even though consultations were longer. PMID- 16707510 TI - An analysis of B cell selection mechanisms in germinal centers. AB - Affinity maturation of antibodies during immune responses is achieved by multiple rounds of somatic hypermutation and subsequent preferential selection of those B cells that express B cell receptors with improved binding characteristics for the antigen. The mechanism underlying B cell selection has not yet been defined. By employing an agent-based model, we show that for physiologically reasonable parameter values affinity maturation can be driven by competition for neither binding sites nor antigen--even in the presence of competing secreted antibodies. Within the tested mechanisms, only clonal competition for T cell help or a refractory time for the interaction of centrocytes with follicular dendritic cells is found to enable affinity maturation while generating the experimentally observed germinal centre characteristics and tolerating large variations in the initial antigen density. PMID- 16707509 TI - Management of miscarriage: expectant, medical, or surgical? Results of randomised controlled trial (miscarriage treatment (MIST) trial). AB - OBJECTIVE: To ascertain whether a clinically important difference exists in the incidence of gynaecological infection between surgical management and expectant or medical management of miscarriage. DESIGN: Randomised controlled trial comparing medical and expectant management with surgical management of first trimester miscarriage. SETTING: Early pregnancy assessment units of seven hospitals in the United Kingdom. PARTICIPANTS: Women of less than 13 weeks' gestation, with a diagnosis of early fetal demise or incomplete miscarriage. INTERVENTIONS: Expectant management (no specific intervention); medical management (vaginal dose of misoprostol preceded, for women with early fetal demise, by oral mifepristone 24-48 hours earlier); surgical management (surgical evacuation). MAIN OUTCOME MEASURES: Confirmed gynaecological infection at 14 days and eight weeks; need for unplanned admission or surgical intervention. RESULTS: 1200 women were recruited: 399 to expectant management, 398 to medical management, and 403 to surgical management. No differences were found in the incidence of confirmed infection within 14 days between the expectant group (3%) and the surgical group (3%) (risk difference 0.2%, 95% confidence interval - 2.2% to 2.7%) or between the medical group (2%) and the surgical group (0.7%, - 1.6% to 3.1%). Compared with the surgical group, the number of unplanned hospital admissions was significantly higher in both the expectant group (risk difference 41%, - 47% to - 36%) and the medical group (- 10%, - 15% to - 6%). Similarly, when compared with the surgical group, the number of women who had an unplanned surgical curettage was significantly higher in the expectant group (risk difference - 39%, - 44% to - 34%) and the medical group (- 30%, - 35% to - 25%). CONCLUSIONS: The incidence of gynaecological infection after surgical, expectant, and medical management of first trimester miscarriage is low (2-3%), and no evidence exists of a difference by the method of management. However, significantly more unplanned admissions and unplanned surgical curettage occurred after expectant management and medical management than after surgical management. TRIAL REGISTRATION NATIONAL RESEARCH REGISTER: N0467011677/N0467073587. PMID- 16707511 TI - Measurement of quality of life in pulmonary hypertension and its significance. AB - Until recently, assessment of patients with pulmonary hypertension has mainly relied on functional and haemodynamic parameters. Health-related quality of life (HRQOL), however, has become increasingly important in defining overall health status. The present study investigated the performance and clinical relevance of the Minnesota Living with Heart Failure (MLHF) questionnaire by prospectively studying 48 patients with either pulmonary arterial hypertension (n = 26) or chronic thromboembolic pulmonary hypertension (n = 22). The MLHF scores were correlated to various clinical and haemodynamic parameters. Prognostic outcome was evaluated by calculating the time taken to reach an adverse clinical event defined as death, lung transplantation or pulmonary endarterectomy. The reliability of test-re-test and internal consistency of this HRQOL tool was high. The MLHF score and its physical subscore correlated moderately to well with functional and haemodynamic parameters, except in the case of pulmonary artery pressures. Both scores significantly improved during vasodilator therapy. This figure was surpassed only by the New York Heart Association/World Health Organization functional class. A multivariate analysis of all variables revealed that the MLHF score was the sole factor predicting subsequent outcome. The Minnesota Living with Heart Failure questionnaire is highly reproducible, consistent, and a moderately valid and responsive tool in assessing health related quality of life in pulmonary hypertension. Moreover, it is a significant predictor of outcome in these patients. PMID- 16707512 TI - Differences in microsatellite DNA level between asthma and chronic obstructive pulmonary disease. AB - Previous studies have shown that microsatellite (MS) DNA instability (MSI) is detectable in sputum cells in chronic obstructive pulmonary disease (COPD) and asthma. The aim of the present study was to investigate whether asthma and COPD could be distinguished at the MS DNA level. DNA was extracted from sputum cells and white blood cells from 63 COPD patients, 60 non-COPD smokers, 36 asthmatics and 30 healthy nonsmokers. Ten MS markers located on chromosomes 2p, 5q, 6p, 10q, 13q, 14q and 17q were analysed. No MSI was detected in non-COPD smokers or healthy nonsmokers. A significantly higher proportion of COPD patients exhibited MSI (49.2%) compared to asthmatics (22.2%). MSI was detected even in the mild stages of COPD (33.3%) and asthma (22.2%). No relationship was found between MSI and COPD severity. The most frequently affected marker was D14S588 (17.5% in COPD and 2.7% in asthma). The markers D6S344, G29802 and D13S71 showed alterations only in COPD, and G29802 was associated with a significantly decreased forced expiratory volume in one second FEV1 (% predicted), whereas MSI in D6S344 was associated with a significantly higher FEV1 (% pred). The frequency of microsatellite instability was higher in chronic obstructive pulmonary disease than in asthma, and microsatellite instability in three workers showed chronic obstructive pulmonary disease specificity. However, further studies are needed to verify the differences between chronic obstructive pulmonary disease and asthma at the microsatellite level. PMID- 16707513 TI - Pine dust, atopy and lung function: A cross-sectional study in sawmill workers. AB - An increased risk of asthma symptoms has previously been shown in 772 pine sawmill workers. The aim of the current study was to assess the association between dust exposure, lung function and atopy. Subjects with (n = 59) and without (n = 167) asthma symptoms were randomly selected from the previous survey. Lung function and atopy were determined using spirometry and skin-prick tests, respectively. Inhalable dust levels were measured on the same day. The geometric mean dust concentration was 0.52 mg x m(-3). Exposure to dry but not to green dust was associated with asthma symptoms. Green dust was associated with atopic sensitisation, particularly against outdoor allergens; no association was found for dry dust. Forced vital capacity, forced expiratory volume in one second and peak expiratory flow were significantly lower in workers exposed to high levels of green dust (-350 mL, -260 mL and -860 mL x s(-1), respectively) and dry dust (-230 mL, -190 mL and -850 mL x s(-1), respectively). These associations were observed both in subjects with and without asthma symptoms. No associations with cross-shift changes in lung function were found. Exposure to green pine sawdust may be a risk factor for atopy. Both green and dry dust were associated with obstructive as well as restrictive pulmonary effects. PMID- 16707514 TI - Increased serum levels of chromogranin A in male smokers with airway obstruction. AB - The neuroendocrine (NE) system may play an important role in smoking-induced airway diseases. The aim of the present study was to examine the relationship between serum levels of the general NE marker chromogranin A (CgA) and smoking habits, respiratory symptoms and lung function. The study population consisted of never-smokers with normal lung function, smokers with normal lung function and smokers with airway obstruction who were randomly selected from the lung study of the Nord-Trondelag Health Study (HUNT). Serum CgA was determined in 151, 138 and 116 subjects, respectively. All subjects were seronegative for Helicobacter pylori. Male smokers with airway obstruction had significantly higher serum CgA levels (median 3.70 nmol x L(-1) (interquartile range 3.10-5.15)) than both smokers with normal lung function (3.00 nmol x L(-1) (2.50-3.67)) and never smokers with normal lung function (2.90 nmol x L(-1) (2.57-3.30)). The elevated levels of CgA correlated with the degree of airway obstruction. Moreover, the presence of respiratory symptoms and chronic bronchitis among male smokers were associated with increased serum CgA levels. Females had CgA levels similar to male smokers independent of smoking status and lung function. Elevated serum chromogranin A levels in subjects with airway obstruction and respiratory symptoms may represent neuroendocrine activation in inflammatory or remodelling processes in the lung. PMID- 16707515 TI - Altered expression of the Smad signalling pathway: implications for COPD pathogenesis. AB - Pulmonary emphysema, as a feature of chronic obstructive pulmonary disease (COPD), is characterised by destruction of alveolar tissue. The present authors previously demonstrated reduced decorin expression in the peribronchial area of COPD patients, reflecting an altered extracellular matrix (ECM) modulation. Decorin transcription is regulated by the transforming growth factor (TGF)-beta Smad pathway, the key intracellular signal route for initiation of ECM component gene transcription. Whether this pathway is aberrantly expressed in COPD is not known. An immunohistochemical study was performed to compare protein expression of TGF-beta1 and TGF-beta receptors, Smad 2, 3, 4 and 7, and decorin in lung tissue of Global Initiative for Chronic Obstructive Lung Disease (GOLD) stages II and IV COPD patients and controls. Epithelial expression of the inhibitory Smad 7 was significantly lower in patients with GOLD stages II and IV than in controls, with other Smad protein expressions being similar in the groups. The expression of TGF-beta1 and TGF-beta receptor type I was significantly lower in stage II patients. Decorin staining of the adventitia and alveolar walls was significantly reduced in COPD stage IV. In conclusion, the transforming growth factor-beta-Smad pathway is aberrantly expressed in chronic obstructive pulmonary disease patients, implying an abnormal tissue repair ultimately resulting in reduced decorin production. The results of the present study contribute to better understanding of the pathogenesis of emphysema and the airway fibrosis observed in chronic obstructive pulmonary disease patients. PMID- 16707516 TI - Predictors of COPD symptoms: does the sex of the patient matter? AB - Although chronic obstructive pulmonary disease (COPD) patients frequently report symptoms, it is not known which factors determine the course of symptoms over time and if these differ according to the sex of the patient. The current study investigated predictors for presence, development and remission of COPD symptoms in 816 males and 312 females completing 3-yr-follow-up in the European Respiratory Society Study on Chronic Obstructive Pulmonary Disease (EUROSCOP). The following were included in generalised estimating equations logistic regression analyses: explanatory variables of treatment; pack-yrs smoking; age, forced expiratory volume in one second % predicted (FEV1 % pred); annual increase in FEV1 and number of cigarettes smoked; body mass index; and phadiatop. Interaction terms of sex multiplied by explanatory variables were tested. Over 3 yrs, similar proportions of males and females reported symptoms. In males only, higher FEV1 % pred was associated with reduction in new symptoms of wheeze and dyspnoea, and symptom prevalence was reduced with annual FEV1 improvement and phlegm prevalence reduced with budesonide treatment (odds ratio 0.66; 95% confidence interval 0.52-0.83). Additionally an increase in the number of cigarettes smoked between visits increased the risk of developing phlegm (1.40 (1.14-1.70)) and wheeze (1.24 (1.03-1.51)) in males but not females. The current study shows longitudinally that symptom reporting is similar by sex. The clinical course of chronic obstructive pulmonary disease can differ by sex, as males show greater response to cigarette exposure and treatment. PMID- 16707517 TI - Prognostic factors for surgical resection in patients with multidrug-resistant tuberculosis. AB - Although surgical lung resection could improve prognosis in some patients with multidrug-resistant tuberculosis (MDR-TB), there are no reports on the optimal candidates for this surgery. The aim of the present study was to elucidate the prognostic factors for surgery in patients with MDR-TB. Patients who underwent lung resection for the treatment of MDR-TB between March 1993 and December 2004 were included in the present study. Treatment failure was defined as greater than or equal to two of the five cultures recorded in the final 12 months of treatment being positive, any one of the final three cultures being positive, or the patient having died during treatment. The variables that affected treatment outcomes were identified through univariate and multivariate logistic regression analysis. In total, 79 patients with MDR-TB were included in the present study. The treatment outcomes of 22 (27.8%) patients were classified as failure. A body mass index <18.5 kg x m(-2), primary resistance, resistance to ofloxacin and the presence of a cavitary lesion beyond the range of the surgical resection were associated with treatment failure. Low body mass index, primary resistance, resistance to ofloxacin and cavitary lesions beyond the range of resection are possible poor prognostic factors for surgical lung resection in multidrug resistant tuberculosis patients. PMID- 16707519 TI - Glaucoma and increased central corneal thickness in aphakic and pseudophakic patients after congenital cataract surgery. AB - AIM: To compare the mean central corneal thickness (CCT) among aphakic and pseudophakic patients following congenital cataract surgery with age matched controls. METHODS: This study included 43 eyes of 43 aphakic and pseudophakic patients following congenital cataract surgery. 44 healthy, age and sex matched volunteers were recruited for comparison with the patients. After a complete eye examination, corneal thickness and intraocular pressure were measured. RESULTS: In the study group, 33 eyes were aphakic, and the remaining 10 eyes were pseudophakic. The median CCT was 556.0 microm (range 490-640 microm) in the control group and 626 microm (range 523-870 microm) in the study group (p<0.05). There was a significant difference in CCT between aphakic and pseudophakic eyes in which an intraocular lens (IOL) had been implanted at the time of congenital cataract surgery (p = 0.011). The same difference was not observed between aphakic and pseudophakic eyes in which an IOL had been implanted secondarily (p = 0.835). The median age of the patients at the time of lensectomy was 24 months (range 1 week to 120 months). There was a negative correlation between the age at lensectomy and CCT (r = -0.485, p = 0.001). CONCLUSION: Aphakic and pseudophakic patients have significantly thicker corneas than age matched controls. This difference can have an important effect on interpreting intraocular pressures in these patients. It is also important to assess the effects of early surgery for congenital cataracts, as well as those of primary and secondary IOL implantation, on CCT. PMID- 16707520 TI - The effects of glaucoma on the latency of the multifocal visual evoked potential. AB - AIMS: To determine the effect of glaucomatous damage on the latency of the multifocal visual evoked potential (mfVEP). METHODS: Monocular mfVEPs were recorded from a glaucoma group (n = 50) defined by a glaucomatous disc and an abnormal visual field and a control group (n = 47). 25 patients were characterised as normal tension glaucoma (NTG) and 25 as high tension glaucoma (HTG). Monocular and interocular latency analyses of the more affected eye were obtained using custom software. RESULTS: On interocular analysis, both the HTG and NTG groups showed a statistically significant increase in mean mfVEP latency with average relative latencies and percentage of points with significant delays of 1.7 ms and 10.3% (HTG) and 1.3 ms and 8.2% (NTG) compared to -0.3 ms and 2.7% (controls). On monocular analysis, only the HTG group showed a significant increase in latency with measures of 5.7 ms and 14.6% (HTG) compared to 3.2 ms and 10.6% (NTG) and 2.1 ms and 9.6% (controls). Using the 95th percentile of a normative group as the cut off, the sensitivity ranged from 20% to 38% and the specificity from 87% to 100% with the interocular analysis providing the best discrimination, CONCLUSION: Although up to 40% of patients showed delays in the mfVEP latency, these delays were modest, on average a few milliseconds. These results differ markedly from those of a recent conventional VEP study, which reported 100% sensitivity, 100% specificity, and an average delay that exceeded 25 ms. PMID- 16707518 TI - Light and inherited retinal degeneration. AB - Light deprivation has long been considered a potential treatment for patients with inherited retinal degenerative diseases, but no therapeutic benefit has been demonstrated to date. In the few clinical studies that have addressed this issue, the underlying mutations were unknown. Our rapidly expanding knowledge of the genes and mechanisms involved in retinal degeneration have made it possible to reconsider the potential value of light restriction in specific genetic contexts. This review summarises the clinical evidence for a modifying role of light exposure in retinal degeneration and experimental evidence from animal models, focusing on retinitis pigmentosa with regional degeneration, Oguchi disease, and Stargardt macular dystrophy. These cases illustrate distinct pathophysiological roles for light, and suggest that light restriction may benefit carefully defined subsets of patients. PMID- 16707521 TI - Incidence of ocular complications in patients with multibacillary leprosy after completion of a 2 year course of multidrug therapy. AB - AIM: To evaluate the incidence of and risk factors for ocular complications in multibacillary (MB) leprosy patients following completion of 2 year, fixed duration, multidrug therapy (MDT). METHODS: Biannual eye examinations were conducted prospectively on a cohort of MB patients who had completed MDT and followed up for 5 years. The incidence of ocular pathology was calculated as the number of events per person year of event free follow up of patients who did not have the specific finding before completion of MDT. RESULTS: 278 patients had one or more follow up visits after completion of MDT. The incidence of lagophthalmos was 0.24%/patient year (95% CI 0.10% to 0.37%); corneal opacity, 5.35%/patient year (95% CI 4.27% to 6.70%); uveal involvement, 3.78%/patient year (95% CI 2.96% to 4.83%); and cataract that reduced vision to 6/18 or less, 2.4%/patient year (95% CI 1.77% to 3.26%). Overall, 5.65%/patient year (95% CI 4.51% to 7.09%) developed leprosy related ocular disease and 3.86%/patient year (95% CI 3.00% to 4.95%) developed leprosy related, potentially blinding ocular pathology during the period following MDT. Age and other disability also predicted incident eye disease. CONCLUSIONS: Every year, approximately 5.6% of patients with MB who have completed MDT can be expected to develop new ocular complications of leprosy, which often (3.9%) are potentially vision threatening. Because many of these complications cannot be detected without slit lamp examination, periodic monitoring, particularly of older patients and those with other disability, is recommended, in order to detect and treat ocular complications satisfactorily. PMID- 16707522 TI - Corneal ulceration in South East Asia. II: a strategy for the prevention of fungal keratitis at the village level in Burma. AB - AIM: To prove that topical antifungal and antibiotic prophylaxis distributed by grass roots village health workers (VHWs) in Burma is an effective public health intervention for the prevention of post-traumatic microbial keratitis in a population where the majority of ulcers are fungal. METHODS: Three villages in Bago District with a combined population of 16,987 were selected for the study. This defined population was followed prospectively for 12 months by 15 VHWs who were trained to identify post-traumatic corneal abrasions with fluorescein dye and a blue torch and to administer 1% chloramphenicol and 1% clotrimazole ointment three times a day for 3 days to the eyes of individuals who fulfilled the eligibility criteria. RESULTS: During the 12 month period 273 individuals reported to VHWs with an ocular injury and 126 were found to have a corneal abrasion. All 126 were treated with 1% chloramphenicol and 1% clotrimazole ointment three times a day for 3 days, and all healed without sequelae. CONCLUSIONS: Both fungal and bacterial ulcers that occur following traumatic corneal abrasions can be effectively prevented in a village setting by using relatively simple measures that local volunteer public health workers can easily be taught to employ. PMID- 16707523 TI - "Worm in the eye": the rationale for treatment of DUSN in south India. AB - AIM: To discuss the rationale for different techniques of treatment for DUSN (diffuse unilateral subacute neuroretinitis) and their effectiveness in two patients from south India. METHODS: Two rare cases of live worms in DUSN from India are reported, where filarial Brugia malayi is endemic. Both cases presented with progressive unilateral loss of vision with no history of animal contact. They were 40 year old, apparently healthy men. In case 1, the worm (1500-2000 microm) was easy to identify with wriggling movements among crisscrossing diffuse subretinal tracks. The worm was destroyed by a single shot of laser to its advancing end, which was followed by oral steroid to control the inflammation caused by the dead worm. In case 2, the worm was small and difficult to identify. Initially diffuse neuroretinitis was diagnosed and treated with intravenous methylprednisolone and oral corticosteroid. A week later, a small live worm (400 600 microm) was found and subsequently destroyed by laser photocoagulation followed by a combination of anthelminthics. RESULTS: The patients' vision had improved to 6/60-6/36 from counting fingers after a few weeks. CONCLUSION: The role of a combination of laser treatment, systemic steroid, and anthelminthics is discussed. PMID- 16707524 TI - Retinoscopic (refractive) estimation of axial length in paediatric aphakia: a comparison with ultrasonic measurement. AB - AIM: To compare paediatric axial length values estimated from the aphakic refraction alone with axial length values measured by ultrasound. METHODS: Retrospective institutional medical record review of paediatric aphakic patients 12 years of age and younger with documented ultrasonic axial length and objective refraction (retinoscopy) within 3 months of each other. An estimate of axial length was made from the aphakic refraction alone (with an assumed average keratometry value of 44 dioptres) for all patients. RESULTS: 149 eyes of 102 paediatric aphakic patients were identified. The ultrasonic axial length values (mean 22.47 mm, SD 1.69, 95% confidence interval (CI) 0.27) and estimated axial length values (mean 22.41 mm, SD 1.53, 95% CI 0.25) had an average difference of 0.05 mm (SD 1.04, 95% CI 0.17) and were not significantly different (p = 0.56) by the two tailed paired t test. A histogram of the differences that did exist between the two values resembled a normal distribution. The nine eyes with the largest differences between the two values had either low hyperopic aphakic refractions or abnormal average keratometry values. CONCLUSIONS: There was no significant difference between the two groups of axial length values, and the distribution of differences that did exist seemed random. The greatest differences between the two values occurred in longer (less hyperopic) eyes and in eyes with abnormally steep or flat keratometry. Estimation of axial length from the aphakic refraction alone seems to be a useful technique in the average paediatric eye, especially if biometry is unavailable. PMID- 16707525 TI - Decreasing efficacy of repeated intravitreal triamcinolone injections in diabetic macular oedema. AB - BACKGROUND/AIM: Intravitreal triamcinolone (IVTA) results in transient improvements in diabetic macular oedema (DMO), necessitating repeated injections. The authors report a case series of 10 eyes of 10 patients with DMO, who received a repeat injection of 4 mg IVTA, at least 26 weeks after the first injection of the same dose. METHOD: Pre-injection and at 2, 4, 9, and 17 weeks post-injection, best corrected visual acuity (BCVA) and central foveal thickness (CFT) on optical coherence tomography, after the first and repeat injections, were compared using paired t test. Side effects were monitored. RESULTS: BCVA, CFT, intraocular pressure (IOP), and cataract scores were not significantly different before initial and repeat injections (given at 32.5 (SD 3.5) weeks after the first injection). Transient improvements of BCVA and CFT were achieved after both injections. However, after the repeat injection, the BCVA was significantly worse at all time points (p<0.05) and so were the best achieved CFT and the CFT at 4 weeks post-injection (p = 0.034 and 0.011 respectively), compared with the initial injection. Post-injection maximum IOPs and increase in cataract scores were not significantly different between the two injections. CONCLUSION: A repeat injection of 4 mg of IVTA may not be as effective as an initial injection for the treatment of DMO. PMID- 16707526 TI - Injectable self inflating hydrogel pellet expanders for the treatment of orbital volume deficiency in congenital microphthalmos: preliminary results with a new therapeutic approach. AB - BACKGROUND/AIM: Children with congenital microphthalmos are usually able to wear an eye prosthesis but the cosmetic aspect is determined by the size of the orbital volume deficiency. Instead of using a ball shaped standard hydrogel expander or a regular orbital implant, which would necessitate enucleation of the microphthalmic eye, this study investigates the feasibility of volume augmentation with injectable pellet expanders, as formerly suggested for acquired anophthalmos in adults only. METHOD: The pellet expander is made from a self inflating hydrogel that takes up water by osmosis (dry state: length 8 mm, diameter 2 mm, volume 0.025 ml; in vitro hydrated state after around 1 day: length 15 mm, diameter 4 mm, volume 0.24 ml; swelling capacity: 9.6-fold). This report concerns six patients (two girls and four boys) aged between 4 months and 42 months with unilateral microphthalmos who were treated by injection of 4-14 pellet expanders into the retrobulbar orbital tissue. Volume augmentation was 1 3.5 ml. The pellets were injected using a customised trocar and placed behind the microphthalmos directed into the intraconal space. RESULTS: The increasing orbital volume was noticeable within 2 days and was confirmed by ultrasonography and magnetic resonance imaging. The final result can be anticipated by the volume augmentation effect produced by the amount of saline solution injected in the orbital apex region. All patients were fitted with an artificial eye, which was subsequently enlarged every 3-5 months. Anophthalmic enophthalmos was fully compensated with this technique. No complications have been encountered to date. CONCLUSIONS: Orbital volume augmentation with injectable self inflating hydrogel expander pellets is apparently a safe, quick, and minimally invasive technique for various indications in orbital reconstructive surgery-for example, to treat an enophthalmic appearance in microphthalmos and congenital or acquired anophthalmos. PMID- 16707527 TI - Chiasmal misrouting and foveal hypoplasia without albinism. AB - BACKGROUND/AIMS: To present the ophthalmological and electrophysiological characteristics of three darkly pigmented, female patients with misrouting and foveal hypoplasia. One of the patients had primary ciliary dyskinesia and situs inversus totalis (Kartagener syndrome). METHODS: Fundus photographs were taken and the angles at which the main temporal arterial branches leave the optic nerve head (ONH) were analysed. Optical coherence tomography (OCT) was performed through the presumed foveal region. Pattern onset visually evoked potentials (VEPs) (check sizes 60', 40/400 ms) were recorded and the chiasmal coefficient was calculated to detect misrouting. RESULTS: Fundus photography showed normally pigmented fundi with absence of the usual foveal hyperpigmentation, foveal avascular zone, and macular and foveal reflexes. On OCT no foveal pit was found. The VEP recordings showed the largest positive CI component over the right hemisphere for the left eye, and over the left hemisphere for the right eye, with the CI almost absent over the ipsilateral hemispheres. The differential derivations showed opposite polarity for the recordings of the two eyes. The chiasmal coefficients of all three patients were significantly indicative of misrouting (-0.99, -0.91, and -0.99, respectively). CONCLUSION: Based on the investigations in these patients the authors propose the hypothesis that foveal hypoplasia and misrouting exist as a distinct entity, and do not comprise the exclusive hallmark of albinism. The findings suggest that misrouting may exert a retrograde influence on foveal development. PMID- 16707528 TI - Vitreous relapse following primary chemotherapy for retinoblastoma: is adjuvant diode laser a risk factor? AB - AIMS: To evaluate rates of vitreous relapse among retinoblastoma patients treated with primary chemotherapy and assess diode laser as a potential risk factor for relapse. METHODS: Retrospective review of all patients treated with primary chemotherapy at a large ocular oncology centre. Eyes that developed vitreous relapse were coded with regard to Reese-Ellsworth Group, laterality, time to relapse, type of relapse (vitreous base or non-vitreous base relapse), treatments used (including adjuvant diode laser), and ocular preservation. Individual tumour foci treated with laser hyperthermia were also coded for laser parameters including power settings, number of treatments, and concomitant administration of systemic chemotherapy (chemothermotherapy). RESULTS: 15 of 106 eyes (14.15%) developed vitreous relapse over a 6 year period. Mean time to relapse was 7.2 months after chemotherapy was completed. Five cases (33%) were of the vitreous base variety. Ocular salvage was attempted in 11 cases using a variety of methods; one patient was lost to follow up. Six of the remaining 10 eyes (60%) were salvaged. Eight of 38 eyes (21%) treated with systemic chemotherapy and laser hyperthermia developed vitreous relapse compared with seven of 68 eyes (10%) treated with primary chemotherapy alone (p<0.005). Laser settings, number of hyperthermia treatments, and the concomitant use of systemic chemotherapy (chemothermotherapy) were not associated with higher rates of vitreous relapse. CONCLUSION: Nearly one in seven eyes with retinoblastoma treated with primary chemotherapy may develop vitreous relapse. The administration of diode laser hyperthermia appears to increase this risk. Despite additional therapy a number of these eyes succumb to enucleation. PMID- 16707529 TI - Integrated monitoring and analysis for early warning of patient deterioration. AB - Recently there has been an upsurge of interest in strategies for detecting at risk patients in order to trigger the timely intervention of a Medical Emergency Team (MET), also known as a Rapid Response Team (RRT). We review a real-time automated system, BioSign, which tracks patient status by combining information from vital signs monitored non-invasively on the general ward. BioSign fuses the vital signs in order to produce a single-parameter representation of patient status, the Patient Status Index. The data fusion method adopted in BioSign is a probabilistic model of normality in five dimensions, previously learnt from the vital sign data acquired from a representative sample of patients. BioSign alerts occur either when a single vital sign deviates by close to +/-3 standard deviations from its normal value or when two or more vital signs depart from normality, but by a smaller amount. In a trial with high-risk elective/emergency surgery or medical patients, BioSign alerts were generated, on average, every 8 hours; 95% of these were classified as 'True' by clinical experts. Retrospective analysis has also shown that the data fusion algorithm in BioSign is capable of detecting critical events in advance of single-channel alerts. PMID- 16707530 TI - Efficacy of nitroglycerin inhalation in reducing pulmonary arterial hypertension in children with congenital heart disease. AB - BACKGROUND: There has been a renewed interest in nitric oxide donor drugs, such as nitroglycerin, delivered by the inhalational route for treatment of pulmonary arterial hypertension (PAH). We investigated the acute effects of inhaled nitroglycerin on pulmonary and systemic haemodynamics in children with PAH associated with congenital heart disease. METHODS: Nineteen children with acyanotic congenital heart disease and a left to right shunt with severe PAH, undergoing routine diagnostic cardiac catheterization were included in this study. Systolic, diastolic and mean systemic as well as pulmonary artery pressures, right atrial pressure and pulmonary capillary wedge pressure (PCWP) were recorded and systemic vascular resistance index (SVRI) and pulmonary vascular resistance index (PVRI) were calculated at room air, following 100% oxygen as well as after nitroglycerin inhalation in all patients. RESULTS: Systolic, diastolic and mean pulmonary artery pressure and PVRI decreased significantly, whereas heart rate, systolic, diastolic and mean systemic arterial pressure, PCWP and SVRI did not change significantly following 100% oxygen or inhalation of nitroglycerin. CONCLUSION: Inhaled nitroglycerin significantly decreases systolic, diastolic and mean pulmonary artery pressure as well as PVRI without affecting systemic haemodynamics, and thus can be used as a therapeutic modality for acute reduction of PAH in children with congenital heart disease. PMID- 16707531 TI - Asporin repeat polymorphism in rheumatoid arthritis. AB - BACKGROUND: Asporin belongs to a family of proteins associated with the cartilage matrix. OBJECTIVE: To investigate the role of the functional polymorphism consisting of an aspartic acid (D) repeat polymorphism located in the ASPN gene in the susceptibility to and clinical outcome of rheumatoid arthritis. METHODS: A total of 803 Spanish Caucasian patients with rheumatoid arthritis and 904 controls of the same ethnic origin and matched for age and sex were included in the study. The asporin D repeat polymorphism was genotyped using polymerase chain reaction with a fluorescent primer. RESULTS: No significant differences were detected in the distribution of the 10 alleles found in our population on comparing patients with rheumatoid arthritis with control groups. Nevertheless, individuals bearing D14 produced rheumatoid factor more often than the rest (85.7% v 72.1%, p = 0.006, odds ratio (OR) = 2.35, 95% confidence interval 1.21 to 4.50), and the mean (SD) onset age was higher in the group of individuals bearing D13 (50.09 (13.94)) compared with the rest (47.21 (14.31)), although the difference did not reach significance (p = 0.06). CONCLUSION: The results do not support a major role for asporin D repeat polymorphism in the susceptibility to rheumatoid arthritis. Nevertheless, they support the influence of this gene on the outcome of the disease. PMID- 16707533 TI - EULAR evidence based recommendations for gout. Part I: Diagnosis. Report of a task force of the Standing Committee for International Clinical Studies Including Therapeutics (ESCISIT). AB - OBJECTIVE: To develop evidence based recommendations for the diagnosis of gout. METHODS: The multidisciplinary guideline development group comprised 19 rheumatologists and one evidence based medicine expert, representing 13 European countries. Ten key propositions regarding diagnosis were generated using a Delphi consensus approach. Research evidence was searched systematically for each proposition. Wherever possible the sensitivity, specificity, likelihood ratio (LR), and incremental cost-effectiveness ratio were calculated for diagnostic tests. Relative risk and odds ratios were estimated for risk factors and co morbidities associated with gout. The quality of evidence was categorised according to the evidence hierarchy. The strength of recommendation (SOR) was assessed using the EULAR visual analogue and ordinal scales. RESULTS: 10 key propositions were generated though three Delphi rounds including diagnostic topics in clinical manifestations, urate crystal identification, biochemical tests, radiographs, and risk factors/co-morbidities. Urate crystal identification varies according to symptoms and observer skill but is very likely to be positive in symptomatic gout (LR = 567 (95% confidence interval (CI), 35.5 to 9053)). Classic podagra and presence of tophi have the highest clinical diagnostic value for gout (LR = 30.64 (95% CI, 20.51 to 45.77), and LR = 39.95 (21.06 to 75.79), respectively). Hyperuricaemia is a major risk factor for gout and may be a useful diagnostic marker when defined by the normal range of the local population (LR = 9.74 (7.45 to 12.72)), although some gouty patients may have normal serum uric acid concentrations at the time of investigation. Radiographs have little role in diagnosis, though in late or severe gout radiographic changes of asymmetrical swelling (LR = 4.13 (2.97 to 5.74)) and subcortical cysts without erosion (LR = 6.39 (3.00 to 13.57)) may be useful to differentiate chronic gout from other joint conditions. In addition, risk factors (sex, diuretics, purine-rich foods, alcohol, lead) and co-morbidities (cardiovascular diseases, hypertension, diabetes, obesity, and chronic renal failure) are associated with gout. SOR for each proposition varied according to both the research evidence and expert opinion. CONCLUSIONS: 10 key recommendations for diagnosis of gout were developed using a combination of research based evidence and expert consensus. The evidence for diagnostic tests, risk factors, and co-morbidities was evaluated and the strength of recommendation was provided. PMID- 16707534 TI - A decision tree for genetic diagnosis of hereditary periodic fever in unselected patients. AB - BACKGROUND: The diagnostic value of molecular analysis of the familial Mediterranean fever (FMF) gene (Mediterranean fever (MEFV)) has been well established only in patients selected on the basis of ethnic background or clinical criteria. Genetic diagnosis for other hereditary periodic fever syndromes has been poorly evaluated. OBJECTIVE: To determine the diagnostic contribution of genetic tests for hereditary periodic syndromes in a large, unselected series of patients. METHODS: A retrospective study was conducted on 1941 patients referred to us for FMF genetic tests between 1997 and 2005. MEFV genotypes were compared with clinical data to appraise criteria for FMF diagnosis. Genetic tests for tumour necrosis factor receptor-associated periodic syndrome (TRAPS), hyperimmunoglobulinaemia D syndrome (HIDS) and cryopyrin associated periodic syndromes (CAPS) were also reviewed. RESULTS: 71% of the 1574 patients with enough data had a clinical diagnosis of FMF according to the widely used Israeli criteria. Two MEFV mutations were found in only 409 patients of this subgroup (sensitivity = 37%) and in 15 (3.3%) of the patients with an improbable clinical diagnosis of FMF (specificity = 97%). Molecular diagnosis for alternate hereditary periodic syndromes was carried out in 456 of the patients having a non conclusive FMF genetic test. A positive diagnosis was obtained in 31 of these patients (TRAPS (n = 19), HIDS (n = 4) and CAPS (n = 8)). CONCLUSIONS: First-line MEFV mutation screening in patients with clinically typical FMF may be appropriate only in particular areas. To optimise genetic diagnosis, we propose a decision tree, which, with the advice of an expert practitioner, could help redirect test indications towards non-FMF hereditary periodic syndromes. PMID- 16707532 TI - EULAR evidence based recommendations for gout. Part II: Management. Report of a task force of the EULAR Standing Committee for International Clinical Studies Including Therapeutics (ESCISIT). AB - OBJECTIVE: To develop evidence based recommendations for the management of gout. METHODS: The multidisciplinary guideline development group comprised 19 rheumatologists and one evidence based medicine expert representing 13 European countries. Key propositions on management were generated using a Delphi consensus approach. Research evidence was searched systematically for each proposition. Where possible, effect size (ES), number needed to treat, relative risk, odds ratio, and incremental cost-effectiveness ratio were calculated. The quality of evidence was categorised according to the level of evidence. The strength of recommendation (SOR) was assessed using the EULAR visual analogue and ordinal scales. RESULTS: 12 key propositions were generated after three Delphi rounds. Propositions included both non-pharmacological and pharmacological treatments and addressed symptomatic control of acute gout, urate lowering therapy (ULT), and prophylaxis of acute attacks. The importance of patient education, modification of adverse lifestyle (weight loss if obese; reduced alcohol consumption; low animal purine diet) and treatment of associated comorbidity and risk factors were emphasised. Recommended drugs for acute attacks were oral non-steroidal anti inflammatory drugs (NSAIDs), oral colchicine (ES = 0.87 (95% confidence interval, 0.25 to 1.50)), or joint aspiration and injection of corticosteroid. ULT is indicated in patients with recurrent acute attacks, arthropathy, tophi, or radiographic changes of gout. Allopurinol was confirmed as effective long term ULT (ES = 1.39 (0.78 to 2.01)). If allopurinol toxicity occurs, options include other xanthine oxidase inhibitors, allopurinol desensitisation, or a uricosuric. The uricosuric benzbromarone is more effective than allopurinol (ES = 1.50 (0.76 to 2.24)) and can be used in patients with mild to moderate renal insufficiency but may be hepatotoxic. When gout is associated with the use of diuretics, the diuretic should be stopped if possible. For prophylaxis against acute attacks, either colchicine 0.5-1 mg daily or an NSAID (with gastroprotection if indicated) are recommended. CONCLUSIONS: 12 key recommendations for management of gout were developed, using a combination of research based evidence and expert consensus. The evidence was evaluated and the SOR provided for each proposition. PMID- 16707535 TI - Diurnal variation in serum levels of cartilage oligomeric matrix protein in patients with knee osteoarthritis or rheumatoid arthritis. AB - OBJECTIVE: To monitor changes in serum concentrations of cartilage oligomeric matrix protein (COMP) during a 24-h period to determine any diurnal variation, and to estimate the half life of COMP in the circulation in patients with symptomatic knee osteoarthritis and in those with rheumatoid arthritis. METHODS: Serum samples were drawn every 4 h (7 samples/patient over 24 h) in 10 patients with knee osteoarthritis and 14 patients with rheumatoid arthritis. Osteoarthritis was defined radiographically and clinically (American College of Rheumatology (ACR) criteria) and rheumatoid arthritis according to the 1987 ACR criteria. Serum COMP was measured by sandwich ELISA. A statistical model for the diurnal variation in the COMP levels was developed using the computer program NONMEM. RESULTS: No considerable changes in COMP levels were observed during the day between 08:00 and 21:00 in either group. A significant decrease in serum COMP was apparent during bed rest at night, reaching the lowest levels between 04:00 and 05:00 (p<0.03 or better v all other time points) in patients with osteoarthritis and in those with rheumatoid arthritis. From the rate of decreasing serum COMP levels, a putative half life of COMP in the circulation was estimated to be 7.4 h. CONCLUSION: During normal daytime activities, serum COMP levels are constant. The decrease during the night indicates a rapid elimination of COMP once it has reached the circulation. The stable COMP levels during the day suggest that it is not necessary to further standardise the time of serum sampling in clinical practice. PMID- 16707538 TI - Routine weighing of babies: does it improve feeding and care? PMID- 16707536 TI - Power Doppler ultrasonography and synovitis: correlating ultrasound imaging with histopathological findings and evaluating the performance of ultrasound equipments. AB - OBJECTIVES: To examine the validity of power Doppler ultrasound imaging to identify synovitis, using histopathology as gold standard, and to assess the performance of ultrasound equipments. METHODS: 44 synovial sites in small and large joints, bursae and tendon sheaths were depicted with ultrasound. A synovial biopsy was performed on the site depicted and a synovial sample was taken for histopathological evaluation. The performance of three ultrasound devices was tested using flow phantoms. RESULTS: A positive Doppler signal was detected in 29 of 35 (83%) of the patients with active histological inflammation. In eight additional samples, histological examination showed other pathological synovial findings and a Doppler signal was detected in five of them. No significant correlation was found between the amount of Doppler signal and histological synovitis score (r = 0.239, p = NS). The amount of subsynovial infiltration of polymorphonuclear leucocytes and surface fibrin correlated significantly with the amount of power Doppler signal: r = 0.397 (p<0.01) and 0.328 (p<0.05), respectively. The ultrasound devices differed in showing the smallest detectable flow. CONCLUSIONS: A negative Doppler signal does not exclude the possibility of synovitis. A positive Doppler signal in the synovium is an indicator of an active synovial inflammation in patients. A Doppler signal does not correlate with the extent of the inflammation and it can also be seen in other synovial reactions. It is important that the quality measurements of ultrasound devices are reported, because the results should be evaluated against the quality of the device used. PMID- 16707537 TI - Benefits of a programme taking advantage of patient-instructors to teach and assess musculoskeletal skills in medical students. AB - AIM: To evaluate a rheumatoid arthritis patient-instructor-based formation assessment programme for its ability to improve and assess musculoskeletal knowledge and skills in third-year medical students. METHODS: (1) The quality of our musculoskeletal teaching was assessed before patient-instructor intervention through an open-questions test (pre-test) and performance record forms (PRFs) filled in by the patient-instructors. (2) The improvement afforded by patient instructors was evaluated through a second (identical) open-questions test (post test). (3) The resulting skills in the students were further assessed by an individual patient-instructors physical status record form (PSRF), filled in by the students. RESULTS: Pre-tests and post-tests showed an improvement in correct answers from a mean score of 39% to 47%. The history-taking questions that obtained <50% scores in the pre-test mostly dealt with the consequences of a chronic illness. Intervention of patient-instructors especially improved knowledge of the psychosocial aspects and side effects of drugs. With regard to physical examination, patient-instructors makedly improved the identification of assessment of signs of active and chronic inflammation. PRF analysis showed that 10 of 28 questions answered by <50% of the students were related to disease characteristics of rheumatoid arthritis, extra-articular signs, side effects of drugs and psychosocial aspects. Analysis of the PSRF indicated that the weakness of our students' physical examination abilities in particular is related to recognising the types of swelling and differentiating tenderness from pain on motion. CONCLUSION: This study proves the considerable benefits of the involvement of patient-instructors in the teaching and assessment of clinical skills in students. PMID- 16707539 TI - Is tympanic membrane thermometry the best method for recording temperature in children? AB - Tympanic membrane thermometry has become increasingly popular for measuring temperature in children. The aim of this review is to ascertain the most appropriate, research-based thermometry method for use with children in acute healthcare settings. The following are considered: Concerns regarding the accuracy of the tympanic membrane thermometer. Whether comparison of the tympanic membrane thermometer with temperature measurement at other body sites is appropriate. How choice of thermometer may be influenced by children's and nurses' preferences, technique, the age of the child, ear size and children with otitis media, perforated eardrums, cerumen (ear wax), ear pain or grommets in situ. Children's preferences and rectal thermometry in relation to children's rights, which have not been reviewed so far. PMID- 16707540 TI - The costs children incur during treatment for cancer: a neglected area in theory and research. AB - This article argues that the costs children incur during treatment for cancer should be rendered visible. As economic agents, children expend time, engage in productive work and participate in activities. Each of these aspects of daily life can be affected by cancer and/or its therapy and can be considered costs of illness. This article challenges researchers to consider eliciting children's views and assessments of these costs to illustrate more fully the impact of cancer on children and their childhoods. The resulting findings will provide a more accurate account of the social and economic costs incurred by families over the course of children's treatment trajectories. Ultimately, such information could be used to lobby for changes which would improve the life circumstances of children and their families. PMID- 16707541 TI - Factors promoting psychological adjustment to childhood atopic eczema. AB - Research has found that children with atopic eczema are more likely to experience psychosocial difficulties than would be expected within the general population. This article aims to explore the relationship between child, parent and family factors in promoting positive adjustment to atopic eczema. Children aged five to 11 years with atopic eczema and their parents were identified from a specialist children's dermatology clinic. Seventy-four respondents completed questionnaires assessing child behaviour, parental well-being and family functioning. Parental psychological health, a supportive family environment and low impact of atopic eczema on family functioning were found to predict lower levels of internalizing behaviour (anxiety, depression and social withdrawal). These findings emphasize the importance of family and parental psychological processes rather than biomedical variables in promoting positive adjustment to atopic eczema. PMID- 16707542 TI - Socio-economic status, smoking during pregnancy and birth outcomes: an analysis of cross-sectional community studies in Liverpool (1993-2001). AB - OBJECTIVE: To assess the influence of socio-economic status on pregnancy outcomes in smoking mothers. STUDY DESIGN: Retrospective analysis. METHODS: Parent completed questionnaires addressing pregnancy smoking patterns and birth outcomes. Were distributed via children aged 5-11 years attending 10 primary schools in Merseyside. Data from three community-based cross-sectional surveys undertaken in 1993, 1998 and 2001 were analysed. Townsend score was used as an indicator of household socioeconomic status. RESULTS: 82.2 percent (n = 3730) of respondents were classified as low (disadvantaged) socio-economic status. One third of mothers smoked during pregnancy and this proportion was significantly higher among disadvantaged than advantaged categories (37.7% versus 14.6%; p < 0.001). The logistic regression analysis showed that only maternal smoking during pregnancy was a significant risk factor for adverse birth outcomes when socioeconomic variables were controlled. CONCLUSION: Birth outcomes were worse in babies of smoking mothers during pregnancy independent of household's socio economic status. PMID- 16707543 TI - Compliance with child accident precautions by child health nurses. AB - The aim of this study was to investigate the extent to which parents follow advice from child health nurses about child accident prevention. The study was performed in one of the 10 districts in the city of Malmo, Sweden. Parents to 90, 10-month old children in southern Sweden participated. These parents completed a questionnaire that prompted responses related to parents' background and socio economic factors as well as questions about any precautions they have taken to decrease hazards to children in their home. Odds ratios, confidence intervals and multiple logistic regression analysis was performed. The results indicated that lower educated parents (i.e. those with 12 years' education or less) and foreign born parents complied with a significantly lower level. These results support the need to increase efforts in child accident prevention work. It is important for child health nurses to concentrate their efforts on foreign parents and on less educated parents. Information should be relayed regularly to these parents in a motivational way, in order to decrease risks of child accidents at home. PMID- 16707544 TI - Changing practice: implementing validated paediatric pain assessment tools. AB - This study attempted to address a lack of evidence-based pain management by implementing validated pain assessment tools across a children's hospital. The method used was action research. The first part of this study where nurses' views of pain tools was elicited has already been reported (Simons and Macdonald, 2004). An action research cycle of negotiation, assessment, diagnosing, planning, action, evaluation and withdrawal was utilized. Data collection occurred at two points in time. Three age-appropriate tools were implemented hospital-wide supported by education and clinical input. A survey of nurses was carried out six months and 12 months post implementation of the tools. At the same time evaluation of the use of the tool was performed. Six months after education and implementation 23 percent of children on 10 wards had a pain tool in use. This had increased to 40 percent six months later. There were many inconsistencies between the replies the nurses gave in relation to their reported use of the tools and the actual use of the tools. PMID- 16707545 TI - Long-term follow-up of homograft function after pulmonary valve replacement in patients with tetralogy of Fallot. AB - AIMS: To analyse the long-term outcomes after pulmonary valve replacement (PVR) in patients with a previous correction for tetralogy of Fallot. METHODS AND RESULTS: In a retrospective study, 158 adult patients with a diagnosis of tetralogy of Fallot, who had undergone a PVR after initial total correction in childhood, were identified from the CONCOR (CONgenital CORvitia) registry. All patients underwent 175 PVRs between June 1986 and June 2005. To analyse the predictors for homograft dysfunction and adverse events (death, reoperations, balloon angioplasty), Cox-regression analysis was performed. Overall freedom from significant homograft dysfunction was 66% after 5 years and 47% after 10 years. We could not identify predictors for combined homograft dysfunction. Event-free survival was 78% at 10 years and 68% at 15 years after PVR. Both early significant pulmonary regurgitation (PR) (HR 6.8, P = 0.017) and pulmonary stenosis (PS) (HR 4.0, P = 0.037) after surgery were associated with adverse events. When analysing direct post-operative PR or PS, we observed that in patients with severe, pre-operative PR, right ventricular aneurysm/patch resection resulted in a lower post-operative PR (mean difference grade 0.38 +/- 0.14, P = 0.01). Less significant post-operative PS was associated with a higher diameter of the homograft (HR 0.37, P = 0.006). CONCLUSION: While 47% of the patients in our study were free from homograft dysfunction at 10 years after PVR, event-free survival after PVR remained fairly good (78%). Significant residual lesions directly after surgery influenced event-free survival. A smaller diameter of the pulmonary homograft and severe pre-surgical PR were related to early homograft dysfunction after surgery. PMID- 16707546 TI - Obesity and cardiovascular events in patients with established coronary disease. AB - AIMS: To explore the association between obesity and major adverse coronary events (MACE) in patients with established coronary artery disease (CAD). METHODS AND RESULTS: The Prevention of Events with Angiotensin Converting Enzyme Inhibition (PEACE) Trial randomized 8290 patients with stable CAD and left ventricular (LV) ejection fraction (EF) (LVEF) > or =0.40 to trandolapril or placebo and followed them for a median of 4.8 years. In PEACE patients who were non-diabetic at baseline (5693 men and 1171 women), we used proportional hazards models to conduct a post hoc analysis to examine whether obesity, defined as a body mass index (BMI) > or =30 kg/m(2), is an independent risk factor for the composite endpoint of MACE, defined as cardiovascular death, non-fatal myocardial infarction, coronary revascularization, or stroke. The analysis was conducted separately for men and women. The baseline prevalence of obesity was 28.5% in men and 28.9% in women. After adjusting for significant confounders, obesity was associated with MACE in men [hazard ratio (HR) = 1.28, 95% CI 1.13-1.46, P < 0.01], but not in women (HR = 0.96, 95% CI 0.70-1.31, P = 0.77). Further categorization of BMI showed a J-shaped association between BMI and MACE in the men, and no association in the women. CONCLUSION: In the presence of established CAD, obesity is associated with risk for MACE in men, but there is no support of an association in women. This finding requires further evaluation. PMID- 16707547 TI - Tako-tsubo syndrome: a form of spontaneous aborted myocardial infarction? PMID- 16707548 TI - Advanced therapy may delay the need for transplantation in patients with the Eisenmenger syndrome. AB - AIMS: Advanced therapies (prostacyclin analogues, endothelin receptor antagonists) are successfully used in the treatment of idiopathic pulmonary arterial hypertension. In addition, patients with the Eisenmenger syndrome (ES) seem to benefit from these news drugs regarding symptoms, but there is still no evidence for changes in outcome. METHODS AND RESULTS: The clinical course of 43 patients (M/F 13/30, age 34.0 +/- 12.7 years), registered with unstable ES in our database, was retrospectively analysed. These patients were divided into two groups: those treated with and those treated without advanced therapy. The primary endpoint was defined as death from any cause. Death or inscription on the active waiting list of heart-lung transplantation was considered as secondary endpoint. Kaplan-Meier survival and log rank testing were performed to determine differences in outcome between the two groups. The total cohort was followed for a median period of 4.9 (range 0.2-14.9) years. Mean survival time for patients treated with (n = 26) and without (n = 17) advanced therapy therapies were 8.5 +/ 1.5 and 8.5 +/- 0.9 years, respectively (log rank testing, P = 0.31). However, the mean time to death or inscription on the active waiting list was significantly longer for patients treated with advanced therapy when compared with those without (7.8 +/- 1.0 vs. 3.4 +/- 0.9 years, P = 0.006). CONCLUSION: For the given follow-up period, no improvement in survival time could be documented in adult patients with unstable ES treated with advanced therapy. However, we might suggest with these data that the need for heart-lung transplantation can be substantially delayed with new drugs. PMID- 16707549 TI - Clinical impact of direct referral to primary percutaneous coronary intervention following pre-hospital diagnosis of ST-elevation myocardial infarction. AB - AIMS: Treatment delay is a powerful predictor of survival in ST-elevation myocardial infarction (STEMI) patients undergoing primary percutaneous coronary intervention (PCI). We investigated effectiveness of pre-hospital diagnosis of STEMI with direct referral to PCI, alongside more conventional referral strategies. METHODS AND RESULTS: From January 2003 to December 2004, 658 STEMI patients were referred for primary PCI at our intervention laboratory. Three predefined referral routes were compared: (1) for patients within 90 min drive of the PCI centre, pre-hospital diagnosis and direct transportation (n=166), (2) diagnosis at the interventional hospital emergency department (n=316), (3) diagnosis at local hospitals before transportation (n = 176). Pre-hospital diagnosis was associated with more than 45 min reduction in treatment delay (P = 0.001). No significant difference in in-hospital mortality was apparent in the overall study population. In the cardiogenic shock subgroup (n = 80), pre hospital diagnosis was associated with a two-thirds reduction in in-hospital mortality (P = 0.019); mortality was only 6.2% in shock patients who underwent PCI in < 2 h. CONCLUSION: This study shows that pre-hospital diagnosis can provide a reduction in primary PCI treatment delay, and suggests the hypothesis that this referral strategy might provide survival benefits to patients with cardiogenic shock. PMID- 16707550 TI - Recurrent cardiovascular events in contemporary cardiology: obesity patients should not rest in PEACE. PMID- 16707551 TI - Comparative effects of a low-carbohydrate diet and exercise plus a low carbohydrate diet on muscle sarcoplasmic reticulum responses in males. AB - We employed a glycogen-depleting session of exercise followed by a low carbohydrate (CHO) diet to investigate modifications that occur in muscle sarcoplasmic reticulum (SR) Ca(2+)-cycling properties compared with low-CHO diet alone. SR properties were assessed in nine untrained males [peak aerobic power (Vo(2 peak)) = 43.6 +/- 2.6 (SE) ml.kg(-1).min(-1)] during prolonged cycle exercise to fatigue performed at approximately 58% Vo(2 peak) after 4 days of low CHO diet (Lo CHO) and after glycogen-depleting exercise plus 4 days of low-CHO (Ex+Lo CHO). Compared with Lo CHO, Ex+Lo CHO resulted in 12% lower (P < 0.05) resting maximal Ca(2+)-ATPase activity (V(max) = 174 +/- 12 vs. 153 +/- 10 micromol.g protein(-1).min(-1)) and smaller reduction in V(max) induced during exercise. A similar effect was observed for Ca(2+) uptake. The Hill coefficient, defined as slope of the relationship between cytosolic free Ca(2+) concentration and Ca(2+)-ATPase activity, was higher (P < 0.05) at rest (2.07 +/- 0.15 vs. 1.90 +/- 0.10) with Ex+Lo CHO, an effect that persisted throughout the exercise. The coupling ratio, defined as the ratio of Ca(2+) uptake to V(max), was 23-30% elevated (P < 0.05) at rest and during the first 60 min of exercise with Ex+Lo CHO. The approximately 27 and 34% reductions (P < 0.05) in phase 1 and phase 2 Ca(2+) release, respectively, observed during exercise with Lo CHO were not altered by Ex+Lo CHO. These results indicate that when prolonged exercise precedes a short-term Lo CHO diet, Ca(2+) sequestration properties and efficiency are improved compared with those during Lo CHO alone. PMID- 16707552 TI - Loss of flow induces leukocyte-mediated MMP/TIMP imbalance in dynamic in vitro blood-brain barrier model: role of pro-inflammatory cytokines. AB - There is substantial evidence linking blood-brain barrier (BBB) failure during cerebral ischemia to matrix metalloproteinases (MMP). BBB function may be affected by loss of shear stress under normoxia/normoglycemia, as during cardiopulmonary bypass procedures. The present study used an in vitro flow perfused BBB model to analyze the individual contributions of flow, cytokine levels, and circulating blood leukocytes on the release/activity of MMP-9, MMP-2, and their endogenous inhibitors, the tissue inhibitors of MMPs (TIMPs), TIMP-1, and TIMP-2. The presence of circulating blood leukocytes under normoxic/normoglycemic flow cessation/reperfusion significantly increased the luminal levels of MMP-9 and activity of MMP-2, accompanied by partial reduction of TIMP-1, complete reduction of TIMP-2 and increased BBB permeability. These changes were not observed during constant flow with circulating blood leukocytes, or after normoxic/normoglycemic or hypoxic/hypoglycemic flow cessation/reperfusion without circulating blood leukocytes. The addition of anti IL-6 or anti-TNF-alpha antibody in the lumen before reperfusion suppressed the levels of MMP-9 and activity of MMP-2, had no effect on TIMP-1, and completely restored TIMP-2 and BBB integrity. Injection of TIMP-2 in the lumen before reperfusion prevented the activation of MMP-2 and BBB permeability. These data indicate that blood leukocytes and loss of flow are major factors in the activation of MMP-2, and that cytokine-mediated differential regulation of TIMP-1 and TIMP-2 may contribute significantly to BBB failure. PMID- 16707553 TI - Adenosine metabolism and cancer. Focus on "Adenosine downregulates DPPIV on HT-29 colon cancer cells by stimulating protein tyrosine phosphatases and reducing ERK1/2 activity via a novel pathway". PMID- 16707554 TI - The human NBCe1-A mutant R881C, associated with proximal renal tubular acidosis, retains function but is mistargeted in polarized renal epithelia. AB - The human electrogenic renal Na-HCO(3) cotransporter (NBCe1-A; SLC4A4) is localized to the basolateral membrane of proximal tubule cells. Mutations in the SLC4A4 gene cause an autosomal recessive proximal renal tubular acidosis (pRTA), a disease characterized by impaired ability of the proximal tubule to reabsorb HCO(3)(-) from the glomerular filtrate. Other symptoms can include mental retardation and ocular abnormalities. Recently, a novel homozygous missense mutant (R881C) of NBCe1-A was reported from a patient with a severe pRTA phenotype. The mutant protein was described as having a lower than normal activity when expressed in Xenopus oocytes, despite having normal Na(+) affinity. However, without trafficking data, it is impossible to determine the molecular basis for the phenotype. In the present study, we expressed wild-type NBCe1-A (WT) and mutant NBCe1-A (R881C), tagged at the COOH terminus with enhanced green fluorescent protein (EGFP). This approach permitted semiquantification of surface expression in individual Xenopus oocytes before assay by two-electrode voltage clamp or measurements of intracellular pH. These data show that the mutation reduces the surface expression rather than the activity of the individual protein molecules. Confocal microscopy on polarized mammalian epithelial kidney cells [Madin-Darby canine kidney (MDCK)I] expressing nontagged WT or R881C demonstrates that WT is expressed at the basolateral membrane of these cells, whereas R881C is retained in the endoplasmic reticulum. In summary, the pathophysiology of pRTA caused by the R881C mutation is likely due to a deficit of NBCe1-A at the proximal tubule basolateral membrane, rather than a defect in the transport activity of individual molecules. PMID- 16707555 TI - ATP and PIP2 dependence of the magnesium-inhibited, TRPM7-like cation channel in cardiac myocytes. AB - The Mg(2+)-inhibited cation (MIC) current (I(MIC)) in cardiac myocytes biophysically resembles currents of heterologously expressed transient receptor potential (TRP) channels, particularly TRPM6 and TRPM7, known to be important in Mg(2+) homeostasis. To understand the regulation of MIC channels in cardiac cells, we used the whole cell voltage-clamp technique to investigate the role of intracellular ATP in pig, rat, and guinea pig isolated ventricular myocytes. I(MIC), studied in the presence or absence of extracellular divalent cations, was sustained for >or=50 min after patch rupture in ATP-dialyzed cells, whereas in ATP-depleted cells I(MIC) exhibited complete rundown. Equimolar substitution of internal ATP by its nonhydrolyzable analog adenosine 5'-(beta,gamma imido)triphosphate failed to prevent rundown. In ATP-depleted cells, inhibition of lipid phosphatases by fluoride + vanadate + pyrophosphate prevented I(MIC) rundown. In contrast, under similar conditions neither the inhibition of protein phosphatases 1, 2A, 2B or of protein tyrosine phosphatase nor the activation of protein kinase A (forskolin, 20 microM) or protein kinase C (phorbol myristate acetate, 100 nM) could prevent rundown. In ATP-loaded cells, depletion of phosphatidylinositol 4,5-bisphosphate (PIP(2)) by prevention of its resynthesis (10 microM wortmannin or 15 microM phenylarsine oxide) induced rundown of I(MIC). Finally, loading ATP-depleted cells with exogenous PIP(2) (10 microM) prevented rundown. These results suggest that PIP(2), likely generated by ATP-utilizing lipid kinases, is necessary for maintaining cardiac MIC channel activity. PMID- 16707556 TI - Bitter stimuli induce Ca2+ signaling and CCK release in enteroendocrine STC-1 cells: role of L-type voltage-sensitive Ca2+ channels. AB - We previously demonstrated the expression of bitter taste receptors of the type 2 family (T2R) and the alpha-subunits of the G protein gustducin (Galpha(gust)) in the rodent gastrointestinal (GI) tract and in GI endocrine cells. In this study, we characterized mechanisms of Ca(2+) fluxes induced by two distinct T2R ligands: denatonium benzoate (DB) and phenylthiocarbamide (PTC), in mouse enteroendocrine cell line STC-1. Both DB and PTC induced a marked increase in intracellular [Ca(2+)] ([Ca(2+)](i)) in a dose- and time-dependent manner. Chelating extracellular Ca(2+) with EGTA blocked the increase in [Ca(2+)](i) induced by either DB or PTC but, in contrast, did not prevent the effect induced by bombesin. Thapsigargin blocked the transient increase in [Ca(2+)](i) induced by bombesin, but did not attenuate the [Ca(2+)](i) increase elicited by DB or PTC. These results indicate that Ca(2+) influx mediates the increase in [Ca(2+)](i) induced by DB and PTC in STC-1 cells. Preincubation with the L-type voltage sensitive Ca(2+) channel (L-type VSCC) blockers nitrendipine or diltiazem for 30 min inhibited the increase in [Ca(2+)](i) elicited by DB or PTC. Furthermore, exposure to the L-type VSCCs opener BAY K 8644 potentiated the increase in [Ca(2+)](i) induced by DB and PTC. Stimulation with DB also induced a marked increase in the release of cholecystokinin from STC-1 cells, an effect also abrogated by prior exposure to EGTA or L-type VSCC blockers. Collectively, our results demonstrate that bitter tastants increase [Ca(2+)](i) and cholecystokinin release through Ca(2+) influx mediated by the opening of L-type VSCCs in enteroendocrine STC-1 cells. PMID- 16707558 TI - Interleukin-6 biology is coordinated by membrane-bound and soluble receptors: role in inflammation and cancer. AB - Cytokine receptors, which exist in membrane-bound and soluble forms, bind their ligands with comparable affinity. Although most soluble receptors are antagonists and compete with their membrane-associated counterparts for the ligands, certain soluble receptors are agonists. In these cases, complexes of ligand and soluble receptor bind on target cells to second receptor subunits and initiate intracellular signaling. The soluble receptors of the interleukin (IL)-6 family of cytokines (sIL-6R, sIL-11R, soluble ciliary neurotrophic factor receptor) are agonists capable of transmitting signals through interaction with the universal signal-transducing receptor for all IL-6 family cytokines, gp130. In vivo, the IL 6/sIL-6R complex stimulates several types of cells, which are unresponsive to IL 6 alone, as they do not express the membrane IL-6R. We have named this process trans-signaling. The generation of soluble cytokine receptors occurs via two distinct mechanisms-limited proteolysis and translation-from differentially spliced mRNA. We have demonstrated that a soluble form of the IL-6 family signaling receptor subunit gp130, which is generated by differential splicing, is the natural inhibitor of IL-6 trans-signaling responses. We have shown that in many chronic inflammatory diseases, including chronic inflammatory bowel disease, peritonitis, rheumatoid arthritis, asthma, as well as colon cancer, IL-6 trans signaling is critically involved in the maintenance of a disease state, by promoting transition from acute to chronic inflammation. Moreover, in all these models, the course of the disease can be disrupted by specifically interfering with IL-6 trans-signaling using the soluble gp130 protein. The pathophysiological mechanisms by which the IL-6/sIL-6R complex regulates the inflammatory state are discussed. PMID- 16707557 TI - A novel role for IL-18 in corticosterone-mediated intestinal damage in a two-hit rodent model of alcohol intoxication and injury. AB - Recent findings from our laboratory have shown that acute alcohol (EtOH) intoxication before burn injury impairs intestinal immunity and barrier functions. To further delineate the mechanism of impaired intestinal barrier function, the present study examined the role of corticosterone (CORT) and interleukin (IL)-18, as CORT and IL-18 are elevated following a combined insult of EtOH intoxication and burn injury. Male rats (approximately 250 g) were gavaged with EtOH to achieve a blood EtOH level of approximately 100 mg/dL prior to burn or sham injury (25% total body surface area). Immediately after injury, a group of rats was treated with CORT synthesis inhibitor metyrapone (25 mg/kg), with or without recombinant (r)IL-18 (50 microg/kg). Another group of rats was treated with caspase-1 inhibitor Ac-YVAD-CHO to block IL-18 production. On Day 1 after injury, there was a significant increase in blood CORT levels, intestinal levels of IL-18, neutrophil chemokines [cytokine-induced neutrophil chemoattractant 1 (CINC-1) and CINC-3], intercellular adhesion molecule-1, myeloperoxidase activity, and intestinal permeability in rats receiving a combined insult of EtOH and burn injury. Treatment of rats with CORT inhibitor or with caspase-1 inhibitor prevented the increase in all of the above parameters following a combined insult of EtOH and burn injury. Moreover, coadministration of rIL-18 in metyrapone-treated rats restored the above parameters, similar to those observed in rats receiving EtOH and burn injury. These findings suggest that a combined insult of EtOH and burn injury results in increased CORT levels, which in turn up-regulates intestinal IL-18 levels and thereby causes altered intestinal barrier function following a combined insult of EtOH intoxication and burn injury. PMID- 16707559 TI - High interleukin-10 production is associated with low antibody response to influenza vaccination in the elderly. AB - The present study was designed to determine the correlation among dehydroepiandrosterone (DHEA), cortisol plasma levels, and immune functionality at the time of vaccination with antibody response to influenza vaccination in young and old, healthy volunteers. Fifty-two elderly subjects, ages 63-85 years, and 14 young subjects, ages 26-41 years, entered the study. Plasma levels of DHEA and cortisol and in vitro cytokine production in response to lipopolysaccharide (LPS) and phytohaemagglutinin (PHA) by peripheral blood leukocytes were assessed at the time of vaccination, and antibody titer was measured before and 18 days after influenza virus vaccination. Elderly subjects were characterized by an increase in the cortisol:DHEA ratio, mainly as a result of a decrease in DHEA. A decrease in LPS-induced tumor necrosis factor alpha (TNF-alpha), increased PHA induced interleukin-10 (IL-10) release, and similar PHA-induced interferon-gamma production were observed in elderly subjects compared with young volunteers. Lower antibody titer to influenza A virus was observed in elderly individuals, and the seroconversion factor was found to be correlated inversely with IL-10 production and correlated directly with TNF-alpha production and to a lesser extent, with the plasma level of DHEA. These results suggest that altered cytokine production in elderly subjects at the moment of vaccination can be predictive of a low response to influenza vaccination and warrant the study of strategies to improve protection afforded by the use of vaccines. PMID- 16707560 TI - Immune phenomena involved in the in vivo regression of fibrosarcoma cells expressing cell-associated IL-1alpha. AB - Constitutive expression of cell-associated, but not secreted, interleukin-1alpha (IL-1alpha) by oncogene-transformed fibrosarcoma cells induced regressing tumors in mice, a phenomenon that was abrogated by the IL-1 inhibitor, the IL-1 receptor antagonist (IL-1Ra). On the contrary, non-IL-1alpha-expressing tumor cells induce progressive tumors in mice. In vivo and ex vivo experiments have shown that regression of IL-1alpha-positive fibrosarcoma cells depends on CD8(+) T cells, which can also be activated in CD4(+) T cell-depleted mice, with some contribution of natural killer cells. In spleens of mice bearing the non-IL 1alpha-expressing fibrosarcoma cells, some early and transient manifestations of antitumor-specific immunity, such as activation of specific proliferating T cells, are evident; however, no development of cytolytic T lymphocytes or other antitumor protective cells could be detected. In spleens of mice bearing the non IL-1alpha-expressing fibrosarcoma cells, the development of early tumor-mediated suppression was observed, and in spleens of mice injected with IL-1alpha-positive fibrosarcoma cells, protective immunity developed in parallel to tumor regression. Treatment of mice bearing violent fibrosarcoma tumors with syngeneic inactivated, IL-1alpha-positive fibrosarcoma cells, at a critical interval after injection of the malignant cells (Days 5-12), induced tumor regression, possibly by potentiating and amplifying transient antitumor cell immune responses or by ablation of tumor-mediated suppression. Membrane-associated IL-1alpha may thus serve as an adhesion molecule, which allows efficient cell-to-cell interactions between the malignant and immune effector cells that bear IL-1Rs and function as a focused cytokine with adjuvant activities at nontoxic, low levels of expression. Our results also point to the potential of using antitumor immunotherapeutic approaches using cell-associated IL-1alpha. PMID- 16707561 TI - A common SCN5A polymorphism attenuates a severe cardiac phenotype caused by a nonsense SCN5A mutation in a Chinese family with an inherited cardiac conduction defect. AB - The SCN5A mutations have been associated with a variety of arrhythmic disorders, including type 3 long QT syndrome (LQT3), Brugada syndrome and inherited cardiac conduction defects. The relationship between genotype and phenotype in SCN5A mutations is complex. Some SCN5A mutations may cause death or severe manifestations in some people and may not cause any symptoms or arrhythmias in others. The causes of these unpredictable clinical manifestations remain incompletely understood. The molecular basis of a four-generation family with cardiac conduction abnormalities was studied and whether variants in the SCN5A gene could account for the cardiac phenotypic variability observed in this family was determined. A novel mutation (W1421X) of SCN5A was identified in a four generation family with cardiac conduction abnormalities and several cases of sudden death. Most family members who carry this W1421X mutation have developed major clinical manifestations or electrocardiographic abnormalities, both of which became more prominent as the patients grew older. However, the 73-year-old grandfather, who carried both the W1421X and R1193Q mutations, had thus far remained healthy and presented with only subtle electrocardiographic abnormalities, whereas most of his offspring, who carried a single mutation (W1421X), had died early or had major disease manifestations. This observation suggests that the R1193Q mutation has a complementary role in alleviating the deleterious effects conferred by W1421X in the function of the SCN5A gene. This report provides a good model to explain the mechanism of penetrance of genetic disorders. PMID- 16707562 TI - Crystal retention in renal stone disease: a crucial role for the glycosaminoglycan hyaluronan? AB - The mechanisms that are involved in renal stone disease are not entirely clear. In this article, the various concepts that have been proposed during the past century are reviewed briefly and integrated into current insights. Much attention is dedicated to hyaluronan (HA), an extremely large glycosaminoglycan that may play a central role in renal stone disease. The precipitation of poorly soluble calcium salts (crystal formation) in the kidney is the inevitable consequence of producing concentrated urine. HA is a major constituent of the extracellular matrix in the renal medullary interstitium and the pericellular matrix of mitogen/stress-activated renal tubular cells. HA is an excellent crystal-binding molecule because of its size, negative ionic charge, and ability to form hydrated gel-like matrices. Crystal binding to HA leads to crystal retention in the renal tubules (nephrocalcinosis) and to the formation of calcified plaques in the renal interstitium (Randall's plaques). It remains to be determined whether one or both forms of renal crystal retention are involved in the development of kidney stones (nephrolithiasis). PMID- 16707563 TI - Update on mechanisms of ischemic acute kidney injury. PMID- 16707564 TI - Kidney side population reveals multilineage potential and renal functional capacity but also cellular heterogeneity. AB - Side population (SP) cells in the adult kidney are proposed to represent a progenitor population. However, the size, origin, phenotype, and potential of the kidney SP has been controversial. In this study, the SP fraction of embryonic and adult kidneys represented 0.1 to 0.2% of the total viable cell population. The immunophenotype and the expression profile of kidney SP cells was distinct from that of bone marrow SP cells, suggesting that they are a resident nonhematopoietic cell population. Affymetrix expression profiling implicated a role for Notch signaling in kidney SP cells and was used to identify markers of kidney SP. Localization by in situ hybridization confirmed a primarily proximal tubule location, supporting the existence of a tubular "niche," but also revealed considerable heterogeneity, including the presence of renal macrophages. Adult kidney SP cells demonstrated multilineage differentiation in vitro, whereas microinjection into mouse metanephroi showed that SP cells had a 3.5- to 13-fold greater potential to contribute to developing kidney than non-SP main population cells. However, although reintroduction of SP cells into an Adriamycin nephropathy model reduced albuminuria:creatinine ratios, this was without significant tubular integration, suggesting a humoral role for SP cells in renal repair. The heterogeneity of the renal SP highlights the need for further fractionation to distinguish the cellular subpopulations that are responsible for the observed multilineage capacity and transdifferentiative and humoral activities. PMID- 16707565 TI - Parathyroid hormone 7-84 induces hypocalcemia and inhibits the parathyroid hormone 1-84 secretory response to hypocalcemia in rats with intact parathyroid glands. AB - Biologic effects of large C-terminal parathyroid hormone (PTH) fragments, opposite to those of N-terminal PTH, have been demonstrated. C-terminal PTH fragments are co-secreted with N-terminal PTH from the parathyroids. The aim of our study was to examine whether C-terminal PTH 7-84 regulates secretion of PTH 1 84 and affects the expression of genes of relevance for parathyroid function, PTH, calcium-sensing receptor (CaR), PTH type 1 receptor (PTHR1), and PTH-related peptide (PTHrP) genes in rat parathyroid glands. PTH 7-84 induced a significant decrease in plasma Ca2+ in rats with intact parathyroid glands. Despite the reduction of plasma Ca2+, no stimulation of PTH 1-84 secretion took place. Furthermore, the PTH 1-84 secretory response to EGTA-induced acute and severe hypocalcemia was significantly inhibited by PTH 7-84. During recovery from hypocalcemia, plasma Ca2+ levels were significantly lower in the PTH 7-84-treated group, as compared with the vehicle group, and at the same time plasma PTH 1-84 levels were significantly suppressed. The expression of PTH, CaR, PTHR1, and PTHrP genes in the rat parathyroid glands was not affected by PTH 7-84. The peripheral metabolism of PTH 1-84 was not affected by PTH 7-84. PTH 7-84 did not cross-react with the rat bioactive PTH 1-84 assay. In normal rats with intact parathyroid glands, PTH 7-84 inhibited the PTH 1-84 secretory response to hypocalcemia and induced a significant decrease in plasma Ca2+. These effects of PTH 7-84 on PTH 1-84 secretion and on plasma Ca2+ levels were not associated with significant changes in PTH, PTHR1, CaR, and PTHrP gene expressions in the rat parathyroid glands. It is hypothesized that PTH 7-84 regulates PTH secretion via an autocrine/paracrine regulatory mechanism. PMID- 16707566 TI - Low doses of ouabain protect from serum deprivation-triggered apoptosis and stimulate kidney cell proliferation via activation of NF-kappaB. AB - It now generally is agreed that Na,K-ATPase, in addition to its role in the maintenance of Na+ and K+ gradients across the cell membrane, plays a role in communicating information from the extracellular environment to intracellular signaling pathways. It was reported recently that interaction between ouabain bound Na,K-ATPase and the 1,4,5-trisphosphate receptor (IP3R) triggers slow calcium oscillations and activation of NF-kappaB. Here it is demonstrated that this signaling pathway can serve to prevent cell death and promote cell growth. Rat renal proximal tubular cells in primary culture first were grown in the presence of 10% serum and then exposed to 0.2% serum for 24 h to induce apoptosis. Serum starvation increased the apoptotic index from 1.21 +/- 0.26 to 14.01 +/- 1.17%. Ouabain in concentrations that did not inhibit Na,K-ATPase activity (1 to 10 nM) completely abolished the apoptotic effect of serum starvation. Ouabain protection from apoptosis was not observed when release of calcium from intracellular stores via the IP3R was prevented. It was shown that the NH2 terminal tail of the Na,K-ATPase alpha subunit plays a key role in ouabain-triggered calcium oscillations. It was found that ouabain did not protect from apoptosis in serum-deprived cells that expressed a mutant Na,K-ATPase alpha subunit with deletion of the NH2 terminal tail. Ouabain exposure (10 nM for 24 h) significantly increased translocation of NF-kappaB from cytoplasm to nucleus. Helenalin, an inhibitor of NF-kappaB, abolished the antiapoptotic effect of ouabain. Ouabain (0.1 to 10 nM) also was found to stimulate proliferation and increase the viability of kidney cells. These effects were abolished when release of calcium via the IP3R was prevented. PMID- 16707567 TI - PAR1b promotes cell-cell adhesion and inhibits dishevelled-mediated transformation of Madin-Darby canine kidney cells. AB - Mammalian Par1 is a family of serine/threonine kinases comprised of four homologous isoforms that have been associated with tumor suppression and differentiation of epithelial and neuronal cells, yet little is known about their cellular functions. In polarizing kidney epithelial (Madin-Darby canine kidney [MDCK]) cells, the Par1 isoform Par1b/MARK2/EMK1 promotes the E-cadherin dependent compaction, columnarization, and cytoskeletal organization characteristic of differentiated columnar epithelia. Here, we identify two functions of Par1b that likely contribute to its role as a tumor suppressor in epithelial cells. 1) The kinase promotes cell-cell adhesion and resistance of E cadherin to extraction by nonionic detergents, a measure for the association of the E-cadherin cytoplasmic domain with the actin cytoskeleton, which is critical for E-cadherin function. 2) Par1b attenuates the effect of Dishevelled (Dvl) expression, an inducer of wnt signaling that causes transformation of epithelial cells. Although Dvl is a known Par1 substrate in vitro, we determined, after mapping the PAR1b-phosphorylation sites in Dvl, that PAR1b did not antagonize Dvl signaling by phosphorylating the wnt-signaling molecule. Instead, our data suggest that both proteins function antagonistically to regulate the assembly of functional E-cadherin-dependent adhesion complexes. PMID- 16707568 TI - Association of Bcl-2 with misfolded prion protein is linked to the toxic potential of cytosolic PrP. AB - Protein misfolding is linked to different neurodegenerative disorders like Alzheimer's disease, polyglutamine, and prion diseases. We investigated the cytotoxic effects of aberrant conformers of the prion protein (PrP) and show that toxicity is specifically linked to misfolding of PrP in the cytosolic compartment and involves binding of PrP to the anti-apoptotic protein Bcl-2. PrP targeted to different cellular compartments, including the cytosol, nucleus, and mitochondria, adopted a misfolded and partially proteinase K-resistant conformation. However, only in the cytosol did the accumulation of misfolded PrP induce apoptosis. Apoptotic cell death was also induced by two pathogenic mutants of PrP, which are partially localized in the cytosol. A mechanistic analysis revealed that the toxic potential is linked to an internal domain of PrP (amino acids 115-156) and involves coaggregation of cytosolic PrP with Bcl-2. Increased expression of the chaperones Hsp70 and Hsp40 prevented the formation of PrP/Bcl-2 coaggregates and interfered with PrP-induced apoptosis. Our study reveals a compartment-specific toxicity of PrP misfolding that involves coaggregation of Bcl-2 and indicates a protective role of molecular chaperones. PMID- 16707569 TI - Distinct roles for Tsg101 and Hrs in multivesicular body formation and inward vesiculation. AB - In mammalian cells, epidermal growth factor (EGF) stimulation promotes multivesicular body (MVB) formation and inward vesiculation within MVB. Annexin 1 is required for EGF-stimulated inward vesiculation but not MVB formation, demonstrating that MVB formation (the number of MVBs/unit cytoplasm) and inward vesiculation (the number of internal vesicles/MVB) are regulated by different mechanisms. Here, we show that EGF-stimulated MVB formation requires the tumor susceptibility gene, Tsg101, a component of the ESCRT (endosomal sorting complex required for transport) machinery. Depletion of Tsg101 potently inhibits EGF degradation and MVB formation and causes the vacuolar domains of the early endosome to tubulate. Although Tsg101 depletion inhibits MVB formation and alters the morphology of the early endosome in unstimulated cells, these effects are much greater after EGF stimulation. In contrast, depletion of hepatocyte growth factor receptor substrate (Hrs) only modestly inhibits EGF degradation, does not induce tubulation of the early endosome, and causes the generation of enlarged MVBs that retain the ability to fuse with the lysosome. Together, these results indicate that Tsg101 is required for the formation of stable vacuolar domains within the early endosome that develop into MVBs and Hrs is required for the accumulation of internal vesicles within MVBs and that both these processes are up-regulated by EGF stimulation. PMID- 16707570 TI - Targeting of transmembrane protein shrew-1 to adherens junctions is controlled by cytoplasmic sorting motifs. AB - We recently identified transmembrane protein shrew-1 and showed that it is able to target to adherens junctions in polarized epithelial cells. This suggested shrew-1 possesses specific basolateral sorting motifs, which we analyzed by mutational analysis. Systematic mutation of amino acids in putative sorting signals in the cytoplasmic domain of shrew-1 revealed three tyrosines and a dileucine motif necessary for basolateral sorting. Substitution of these amino acids leads to apical localization of shrew-1. By applying tannic acid to either the apical or basolateral part of polarized epithelial cells, thereby blocking vesicle fusion with the plasma membrane, we obtained evidence that the apically localized mutants were primarily targeted to the basolateral membrane and were then redistributed to the apical domain. Further support for a postendocytic sorting mechanism of shrew-1 was obtained by demonstrating that mu1B, a subunit of the epithelial cell-specific adaptor complex AP-1B, interacts with shrew-1. In conclusion, our data provide evidence for a scenario where shrew-1 is primarily delivered to the basolateral membrane by a so far unknown mechanism. Once there, adaptor protein complex AP-1B is involved in retaining shrew-1 at the basolateral membrane by postendocytic sorting mechanisms. PMID- 16707571 TI - The detection of micromolar pericellular ATP pool on lymphocyte surface by using lymphoid ecto-adenylate kinase as intrinsic ATP sensor. AB - Current models of extracellular ATP turnover include transient release of nanomolar ATP concentrations, triggering of signaling events, and subsequent ectoenzymatic inactivation. Given the high substrate specificity for adenylate kinase for reversible reaction (ATP + AMP <--> 2ADP), we exploited lymphoid ecto adenylate kinase as an intrinsic probe for accurate sensing pericellular ATP. Incubation of leukemic T- and B-lymphocytes with [3H]AMP or [alpha-32P]AMP induces partial nucleotide conversion into high-energy phosphoryls. This "intrinsic" AMP phosphorylation occurs in time- and concentration-dependent fashions via nonlytic supply of endogenous gamma-phosphate-donating ATP, remains relatively resistant to bulk extracellular ATP scavenging by apyrase, and is diminished after lymphocyte pretreatment with membrane-modifying agents. This enzyme-coupled approach, together with confocal imaging of quinacrine-labeled ATP stores, suggests that, along with predominant ATP accumulation within cytoplasmic granules, micromolar ATP concentrations are constitutively retained on lymphoid surface without convection into bulk milieu. High basal levels of inositol phosphates in the cells transfected with ATP-selective human P2Y2-receptor further demonstrate that lymphocyte-surrounding ATP is sufficient for triggering purinergic responses both in autocrine and paracrine fashions. The ability of nonstimulated lymphocytes to maintain micromolar ATP halo might represent a novel route initiating signaling cascades within immunological synapses and facilitating leukocyte trafficking between the blood and tissues. PMID- 16707572 TI - Three-dimensional culture regulates Raf-1 expression to modulate fibronectin matrix assembly. AB - Oncogenic transformation has been associated with decreased fibronectin (FN) matrix assembly. For example, both the HT-1080 fibrosarcoma and MAT-LyLu cell lines fail to assemble a FN matrix when grown in monolayer culture (2-dimensional [2D] system). In this study, we show that these cells regain the ability to assemble a FN matrix when they are grown as aggregates (3-dimensional [3D] system). FN matrix assembly in 3D correlates with decreased Raf-1 protein expression compared with cells grown in monolayer culture. This effect is associated with reduced Raf-1 mRNA levels as determined by quantitative RT-PCR and not proteasome-mediated degradation of endogenous Raf-1. Interestingly, transient expression of a Raf-1 promoter-reporter construct demonstrates increased Raf-1 promoter activity in 3D, suggesting that the transition to 3D culture may modulate Raf-1 mRNA stability. Finally, to confirm that decreased Raf 1 expression results in increased FN matrix assembly, we used both pharmacological and small interfering RNA knockdown of Raf-1. This restored the ability of cells in 2D culture to assemble a FN matrix. Moreover, overexpression of Raf-1 prevented FN matrix assembly by cells cultured in 3D, resulting in decreased aggregate compaction. This work provides new insight into how the cell microenvironment may influence Raf-1 expression to modulate cell-FN interactions in 3D. PMID- 16707573 TI - Comprehensive mutation identification in an evolved bacterial cooperator and its cheating ancestor. AB - Precise characterization of the mutation histories of evolutionary lineages is crucial for understanding the evolutionary process, yet mutation identification has been constrained by traditional techniques. We sought to identify all accumulated mutations in an experimentally evolved lineage of the cooperative bacterium Myxococcus xanthus, which constructs fruiting bodies by a process of social multicellular development in response to starvation. This lineage had undergone two major transitions in social phenotype: from an ancestral cooperator to a socially defective cheater, and from the cheater to a competitively dominant cooperator that re-evolved social and developmental proficiency. The 9.14-Mb genome of the evolved, dominant cooperator (strain "PX") was sequenced to approximately 19-fold coverage by using recent "sequencing-by-synthesis" technology and partially sequenced (approximately 45%) by using capillary technology. The resulting data revealed 15 single-nucleotide mutations relative to the laboratory ancestor of PX after the two phases of experimental evolution but no evidence of duplications, transpositions, or multiple-base deletions. No mutations were identified by capillary sequencing beyond those found by pyrosequencing, resulting in a high probability that all mutations were discovered. Seven errors in the reference strain previously sequenced by the Sanger approach were revealed, as were five mutational differences between two distinct laboratory stocks of the reference strain. A single mutation responsible for the restoration of development in strain PX was identified, whereas 14 mutations occurred during the prior phase of experimental evolution. These results provide insight into the genetic basis of two large adaptive transitions in a social bacterium. PMID- 16707575 TI - Oxysterols stimulate Sonic hedgehog signal transduction and proliferation of medulloblastoma cells. AB - Sterol synthesis is required for Sonic hedgehog (Shh) signal transduction. Errors in Shh signal transduction play important roles in the formation of human tumors, including medulloblastoma (MB). It is not clear which products of sterol synthesis are necessary for Shh signal transduction or how they act. Here we show that cholesterol or specific oxysterols are the critical products of sterol synthesis required for Shh pathway signal transduction in MB cells. In MB cells, sterol synthesis inhibitors reduce Shh target gene transcription and block Shh pathway-dependent proliferation. These effects of sterol synthesis inhibitors can be reversed by exogenous cholesterol or specific oxysterols. We also show that certain oxysterols can maximally activate Shh target gene transcription through the Smoothened (Smo) protein as effectively as the known Smo full agonist, SAG. Thus, sterols are required and sufficient for Shh pathway activation. These results suggest that oxysterols may be critical regulators of Smo, and thereby Shh signal transduction. Inhibition of Shh signaling by sterol synthesis inhibitors may offer a novel approach to the treatment of MB and other Shh pathway-dependent human tumors. PMID- 16707574 TI - Inhibition of dsRNA-induced signaling in hepatitis C virus-infected cells by NS3 protease-dependent and -independent mechanisms. AB - The recent establishment of a robust hepatitis C virus (HCV) cell culture system permits analysis of virus-host interactions during HCV infection. Here, we report that HCV genotype 2a (JFH-1) infection fails to induce IFN-beta or IFN-stimulated gene expression in Huh-7 cells, and that it blocks IFN-beta and IFN-stimulated gene production after transfection of synthetic dsRNA. Overexpression of individual components of the dsRNA-signaling pathway in HCV-infected and uninfected cells indicates that HCV inhibits IFN-beta promoter activity by inactivating the mitochondrial antiviral signaling protein/IFN-beta promoter stimulator 1 (MAVS/IPS-1), while leaving the IFN-induced Janus kinases-signal transducers and activators of transcription (JAK-STAT) signaling pathway intact. We also show that MAVS/IPS-1-dependent IFN-beta promoter activity in HCV-infected cells is fully restored by the nonstructural protein 3 (NS3) protease inhibitor BILN2061. In contrast, synthetic dsRNA-induced IFN-beta promoter activity is not restored by BILN2061, although it is partially restored by overexpression of RIG I. These results support recently reported evidence that the HCV NS3 protease blunts the ability of HCV to induce IFN-beta promoter activity by proteolytically cleaving MAVS/IPS-1. The results also suggest that HCV blocks the synthetic dsRNA induced signaling pathway at a point upstream of MAVS/IPS-1, and that it does so by an NS3-independent mechanism. PMID- 16707576 TI - Locating active-site hydrogen atoms in D-xylose isomerase: time-of-flight neutron diffraction. AB - Time-of-flight neutron diffraction has been used to locate hydrogen atoms that define the ionization states of amino acids in crystals of D-xylose isomerase. This enzyme, from Streptomyces rubiginosus, is one of the largest enzymes studied to date at high resolution (1.8 A) by this method. We have determined the position and orientation of a metal ion-bound water molecule that is located in the active site of the enzyme; this water has been thought to be involved in the isomerization step in which D-xylose is converted to D-xylulose or D-glucose to D fructose. It is shown to be water (rather than a hydroxyl group) under the conditions of measurement (pH 8.0). Our analyses also reveal that one lysine probably has an -NH(2)-terminal group (rather than NH(3)(+)). The ionization state of each histidine residue also was determined. High-resolution x-ray studies (at 0.94 A) indicate disorder in some side chains when a truncated substrate is bound and suggest how some side chains might move during catalysis. This combination of time-of-flight neutron diffraction and x-ray diffraction can contribute greatly to the elucidation of enzyme mechanisms. PMID- 16707578 TI - Attachment of capsular polysaccharide to the cell wall of Streptococcus pneumoniae type 2 is required for invasive disease. AB - The capacity of Streptococcus pneumoniae to produce capsular polysaccharide (CPS) is essential for virulence. The CPS biosynthesis proteins CpsB, CpsC, and CpsD function to regulate CPS production via tyrosine phosphorylation of CpsD. This mechanism of regulating CPS production is important for enabling S. pneumoniae to cause invasive disease. Here, we identify mutations affecting the attachment of CPS to the cell wall. These mutations were located in cpsC, such that CpsC functioned independently from CpsD tyrosine phosphorylation. These mutants produced WT levels of CPS, but were unable to cause bacteremia in mice after intranasal challenge. This finding suggests that cell-wall attachment of CPS is essential for invasive pneumococcal disease; production of WT levels of CPS alone is not sufficient. We also show that cpsB mutants, which lack the phosphotyrosine protein phosphatase, produced less CPS than the WT strain, but attached substantially more CPS to their cell wall. Thus, the phosphorylated form of CpsD promotes attachment of CPS to the cell wall. PMID- 16707579 TI - Flows driven by flagella of multicellular organisms enhance long-range molecular transport. AB - Evolution from unicellular organisms to larger multicellular ones requires matching their needs to the rate of exchange of molecular nutrients with the environment. This logistic problem poses a severe constraint on development. For organisms whose body plan is a spherical shell, such as the volvocine green algae, the current (molecules per second) of needed nutrients grows quadratically with radius, whereas the rate at which diffusion alone exchanges molecules grows linearly, leading to a bottleneck radius beyond which the diffusive current cannot meet metabolic demands. By using Volvox carteri, we examine the role that advection of fluid by the coordinated beating of surface-mounted flagella plays in enhancing nutrient uptake and show that it generates a boundary layer of concentration of the diffusing solute. That concentration gradient produces an exchange rate that is quadratic in the radius, as required, thus circumventing the bottleneck and facilitating evolutionary transitions to multicellularity and germ-soma differentiation in the volvocalean green algae. PMID- 16707580 TI - The delivery of copper for thylakoid import observed by NMR. AB - The thylakoid compartments of plant chloroplasts are a vital destination for copper. Copper is needed to form holo-plastocyanin, which must shuttle electrons between photosystems to convert light into biologically useful chemical energy. Copper can bind tightly to proteins, so it has been hypothesized that copper partitions onto ligand-exchange pathways to reach intracellular locations without inflicting damage en route. The copper metallochaperone Atx1 of chloroplast related cyanobacteria (ScAtx1) engages in bacterial two-hybrid interactions with N-terminal domains of copper-transporting ATPases CtaA (cell import) and PacS (thylakoid import). Here we visualize copper delivery. The N-terminal domain PacS(N) has a ferredoxin-like fold that forms copper-dependent heterodimers with ScAtx1. Removal of copper, by the addition of the cuprous-ion chelator bathocuproine disulfonate, disrupts this heterodimer, as shown from a reduction of the overall tumbling rate of the protein mixture. The NMR spectral changes of the heterodimer versus the separate proteins reveal that loops 1, 3, and 5 (the carboxyl tail) of the ScAtx1 Cu(I) site switch to an apo-like configuration in the heterodimer. NMR data ((2)J(NH) couplings in the imidazole ring of (15)N ScAtx1 His-61) also show that His-61, bound to copper(I) in [Cu(I)ScAtx1](2), is not coordinated to copper in the heterodimer. A model for the PacS(N)/Cu(I)/ScAtx1 complex is presented. Contact with PacS(N) induces change to the ScAtx1 copper-coordination sphere that drives copper release for thylakoid import. These data also elaborate on the mechanism to keep copper(I) out of the ZiaA(N) ATPase zinc sites. PMID- 16707581 TI - Regulation of insect steroid hormone biosynthesis by innervating peptidergic neurons. AB - In insects, steroid hormones named ecdysteroids elicit molting and metamorphosis. The prothoracic gland (PG) is a predominant source of ecdysteroids, where their biosynthesis (ecdysteroidogenesis) is regulated by several neuropeptides. Here, we report that FMRFamide-related peptides (FaRPs) regulate ecdysteroidogenesis through direct innervation of the PG in the silkworm Bombyx mori. We purified a previously uncharacterized Bombyx FaRP, DPSFIRFamide, and identified the corresponding Bombyx FMRFamide gene (Bommo-FMRFamide, BRFa), which encodes three additional FaRPs. All BRFa peptides suppressed ecdysteroidogenesis in the PG by reducing cAMP production by means of the receptor for Bommo-myosuppressin, another FaRP we have previously shown to act as a prothoracicostatic factor. BRFa is predominantly expressed in neurosecretory cells of thoracic ganglia, and the neurons in the prothoracic ganglion innervate the PG to supply all four peptides to the gland surface. Electrophysiological recordings during development confirmed the increased firing activity of BRFa neurons in stages with low PG activity and decreased ecdysteroid levels in the hemolymph. To our knowledge, this study provides the first report of peptides controlling ecdysteroidogenesis by direct innervation. PMID- 16707583 TI - Critical bifurcation of shallow microtidal landforms in tidal flats and salt marshes. AB - Shallow tidal basins are characterized by extensive tidal flats and salt marshes that lie within specific ranges of elevation, whereas intermediate elevations are less frequent in intertidal landscapes. Here we show that this bimodal distribution of elevations stems from the characteristics of wave-induced sediment resuspension and, in particular, from the reduction of maximum wave height caused by dissipative processes in shallow waters. The conceptual model presented herein is applied to the Venice Lagoon, Italy, and demonstrates that areas at intermediate elevations are inherently unstable and tend to become either tidal flats or salt marshes. PMID- 16707582 TI - Circadian rhythms in gene transcription imparted by chromosome compaction in the cyanobacterium Synechococcus elongatus. AB - In the cyanobacterium Synechococcus elongatus (PCC 7942) the kai genes A, B, and C and the sasA gene encode the functional protein core of the timing mechanism essential for circadian clock regulation of global gene expression. The Kai proteins comprise the central timing mechanism, and the sensor kinase SasA is a primary transducer of temporal information. We demonstrate that the circadian clock also regulates a chromosome compaction rhythm. This chromosome compaction rhythm is both circadian clock-controlled and kai-dependent. Although sasA is required for global gene expression rhythmicity, it is not required for these chromosome compaction rhythms. We also demonstrate direct control by the Kai proteins on the rate at which the SasA protein autophosphorylates. Thus, to generate and maintain circadian rhythms in gene expression, the Kai proteins keep relative time, communicate temporal information to SasA, and may control access to promoter elements by imparting rhythmic chromosome compaction. PMID- 16707584 TI - State of HIV treatment: Results of the International Association of Physicians in AIDS Care surveys of HIV-positive patients and HIV-treating physicians in the United States. AB - BACKGROUND: There are few reports about physician and patient attitudes about antiretroviral therapy. Reports on physician perceptions of HIV-positive patients and patients' perceptions of their physicians are even scarcer. METHODS: The International Association of Physicians in AIDS Care conducted surveys of HIV treating physicians, and a separate set of HIV-positive patients. Physicians completed an online questionnaire. Patients completed a written questionnaire. RESULTS: Physicians and patients agreed on several issues, including the priority of viral suppression when making treatment decisions and the treatment-limiting impact of side effects. However, they had diverging treatment goals in mind and differing impressions of the type and incidence of side effects. There were also sharp differences in physicians' estimates of how well patients understand HIV disease and its treatment. CONCLUSIONS: The differences revealed through these surveys underline the need to conduct a systematic study of physician and patient attitudes about antiretroviral therapy, as well as physician-patient communication. PMID- 16707577 TI - Evolution of hepatitis C viral quasispecies and hepatic injury in perinatally infected children followed prospectively. AB - Perinatal infection with hepatitis C virus (HCV) is characterized by a wide range of alanine aminotransferase (ALT) levels. The mechanisms responsible for this variability are unknown. We examined whether the evolution of the HCV quasispecies was associated with different ALT profiles in perinatally infected children. Sequences within HCV envelope 1 and 2 genes, inclusive of the hypervariable region 1, the viral load, and the nascent humoral immunity were analyzed in serial serum samples from 12 perinatally infected children prospectively followed for a median of 53 months. These patients were selected to represent two different ALT patterns during the first year of life: 6 had high levels (maximum values ranging from 4.2 to 30 times the normal upper limit), and 6 had normal or slightly elevated levels (< 2 times the normal upper limit). Two patterns of viral evolution were identified according to the ALT profiles. Biochemical evidence of hepatic injury was invariably associated with a mono- or oligoclonal viral population, whereas mild or no liver damage correlated with the early emergence of a heterogeneous viral quasispecies. Consistent with selective immune pressure, amino acid changes occurred almost exclusively within the hypervariable region 1 and were temporally associated with antibody seroconversion; at this time, the difference in genetic diversity between the two groups was highly significant (P = 0.002). The two patterns of viral evolution persisted over time and did not correlate with viral load or genotype. Our study demonstrates that, in perinatally infected children, the evolution of HCV quasispecies correlates with hepatic injury. The sequences reported in this paper have been deposited in the GenBank database (accession nos. DQ 504441-DQ 507112). PMID- 16707586 TI - Agonists of the peroxisome proliferator-activated receptor alpha induce a fiber type-selective transcriptional response in rat skeletal muscle. AB - In rodents, treatment with peroxisome proliferator-activated receptor alpha (PPARalpha) agonists results in peroxisome proliferation, hepatocellular hypertrophy, and hepatomegaly. Drugs in the fibrate class of PPARalpha agonists have also been reported to produce rare skeletal muscle toxicity. Although target driven hepatic effects of PPARalpha treatment have been extensively studied, a characterization of the transcriptional effects of this nuclear receptor/transcription factor on skeletal muscle responses has not been reported. In this study we investigated the effects of PPARalpha agonists on skeletal muscle gene transcription in rats. Further, since statins have been reported to preferentially effect type II muscle fibers, we compared PPARalpha signaling effects between type I and type II muscles. By comparing the transcriptional responses of agonists that signal through different nuclear receptors and using a selection/deselection analytical strategy based on ANOVA, we identified a PPARalpha activation signature that is evident in type I (soleus), but not type II (quadriceps femoris), skeletal muscle fibers. The fiber-type-selective nature of this response is consistent with increased fatty acid uptake and beta oxidation, which represent the major clinical benefits of the hypolipidemic compounds used in this study, but does not reveal any obvious off-target pathways that may drive adverse effects. PMID- 16707587 TI - Coming unglued. PMID- 16707589 TI - Erlotinib: optimizing therapy with predictors of response? PMID- 16707588 TI - United States Food and Drug Administration approval summary: bortezomib for the treatment of progressive multiple myeloma after one prior therapy. AB - PURPOSE: On March 25, 2005, bortezomib (Velcade for Injection; Millennium Pharmaceuticals, Inc., Cambridge, MA, and Johnson & Johnson Pharmaceutical Research & Development, L.L.C.) received regular approval from the U.S. Food and Drug Administration (U.S. FDA) for the treatment of multiple myeloma (MM) progressing after at least one prior therapy. This approval was based on bortezomib's efficacy and safety which was shown in a single, large, comparative international open-label phase 3 trial that randomized 669 patients with MM previously treated with at least one systemic regimen to receive single-agent bortezomib or high-dose dexamethasone. The FDA analysis of the trial data and bortezomib's regulatory development are summarized here. EXPERIMENTAL DESIGN AND RESULTS: Following a preplanned interim analysis of time to disease progression (the primary end point), an independent data-monitoring committee advised the sponsor to halt the study and offer bortezomib to all dexamethasone-treated study patients. Time to progression was significantly prolonged in the bortezomib treatment arm (median, 6.2 months) compared with the dexamethasone arm (median, 3.5 months; log-rank test, P < 0.0001; hazard ratio, 0.55; 95% confidence interval, 0.44-0.69). Analysis of overall survival done on the interim database (with 20% of events) showed the superiority of bortezomib for patients (log-rank test, P < 0.05; hazard ratio, 0.57; 95% confidence interval, 0.40-0.81). Using criteria from the European Group for Blood and Marrow Transplantation, the response rate (complete plus partial response) with bortezomib was also superior to dexamethasone (38% versus 18%; P < 0.0001). Adverse events on the bortezomib arm were similar to those previously observed in phase 2 studies; some notable adverse events included asthenia, peripheral neuropathy, thrombocytopenia, and neutropenia. CONCLUSIONS: The U.S. FDA had earlier (May 2003) granted bortezomib accelerated approval for the treatment of patients with MM progressing after two prior therapies. The results of the phase 3 trial and the FDA analysis of the data, along with the sponsor's completion of other postmarketing commitments, confirm bortezomib's benefit and support regular approval. PMID- 16707590 TI - Practicalities of drugging the phosphatidylinositol-3-kinase/Akt cell survival signaling pathway. PMID- 16707592 TI - RNA interference suppression of MUC1 reduces the growth rate and metastatic phenotype of human pancreatic cancer cells. AB - MUC1 is a highly glycosylated, type I transmembrane protein expressed by normal ductal epithelial cells of the pancreas, breast, lung, and gastrointestinal tract, and overexpressed in many cases of adenocarcinoma. We down-regulated MUC1 expression by RNA interference and investigated the effects on malignant and metastatic potential of a human pancreatic cancer cell line, S2-013. MUC1 suppressed clones, S2-013.MTII.C1 and S2-013.MTII.C2, were established by targeting a sequence 3,151 bp from the initiation codon and characterized in vitro for proliferation, invasion, and adhesion. We evaluated the effects of MUC1 suppression in vivo on tumor growth and metastatic properties following implantation into the cecum or pancreas of athymic mice. MUC1-suppressed clones showed significantly decreased proliferation in vitro and in vivo. Global gene expression was evaluated by oligonucleotide microarray analysis. Surprisingly, genes predicted to increase doubling times (cyclin B1 and cyclin D3) were overexpressed in MUC1-suppressed clones. There were alterations in expression of several genes that may affect the malignant properties of pancreatic cancer. Adhesion of MUC1-suppressed cells in vitro to type IV collagen and fibronectin was slightly increased, and adhesion was slightly decreased to type I collagen and laminin. Results of implantation to cecum and pancreas showed significant reduction of metastasis to lymph nodes, lung, or peritoneal sites compared with S2-013.gfp-neo control cells. These results support the hypothesis that MUC1 contributes significantly to growth and metastasis, and that down-regulation of MUC1 protein expression decreases the metastatic potential of pancreatic adenocarcinoma. PMID- 16707591 TI - Efficient presentation of naturally processed HLA class I peptides by artificial antigen-presenting cells for the generation of effective antitumor responses. AB - Appropriate presentation of tumor-associated antigens (TAA) by antigen-presenting cells (APC) is required for the development of clinically relevant antitumor T cell responses. One common approach, which uses APC pulsed with synthetic peptides, can sometimes generate ineffective immune responses. This failure may, in part, be attributed to the formation of HLA/synthetic pulsed peptide complexes that possess different conformations compared with those of endogenously presented peptides. In addition, endogenous peptides may undergo post translational modifications, which do not occur with synthetic peptides. Because our goal is to induce immunity that can recognize TAA that are endogenously presented by tumors, we designed an APC that would not only express the required immunoaccessory molecules but also naturally process and present target antigenic peptides. In this study, we generated an artificial APC (aAPC) that can endogenously present any chosen HLA-A*0201 (A2)-restricted peptide by processing a fusion protein that contains a unique "LTK" sequence linked to the antigenic peptide. Proteasome-dependent processing is so effective that the presented peptide can be directly eluted from the cell surface and identified by biochemical methods. Furthermore, we found that aAPC, engineered to endogenously present peptide derived from the melanoma antigen MART1, can be used to prime and expand antitumor CTL that target MART1-expressing tumor cells in a HLA-A2 restricted manner. Our engineered aAPC could serve as an "off-the-shelf" APC designed to constitutively express class I-restricted TAA peptides and could be used to generate effective T-cell responses to treat human disease. PMID- 16707593 TI - Mixed lineage leukemia-rearranged childhood pro-B and CD10-negative pre-B acute lymphoblastic leukemia constitute a distinct clinical entity. AB - PURPOSE: Mixed lineage leukemia (MLL) abnormalities occur in approximately 50% of childhood pro-B acute lymphoblastic leukemia (ALL). However, the incidence and type of MLL rearrangements have not been determined in common ALL (cALL) and CD10+ or CD10- pre-B ALL. EXPERIMENTAL DESIGN: To address this question, we analyzed 29 patients with pro-B ALL, 11 patients with CD10- pre-B ALL, 23 pre-B, and 26 cALL patients with CD10 on 20% to 80%, as well as 136 pre-B and 143 cALL patients with CD10 > or = 80% of blasts. They were all enrolled in four Austrian ALL multicenter trials. Conventional cytogenetics were done to detect 11q23 abnormalities and in parallel the potential involvement of the MLL gene was evaluated with a split apart fluorescence in situ hybridization probe set. RESULTS: We found that 15 of 29 pro-B ALL, 7 of 11 CD10- pre-B ALL, and 1 of 2 French-American-British classification L1 mature B-cell leukemia cases had a MLL rearrangement. However, no 11q23/MLL translocation was identified among the CD10+ pre-B and cALL patients. MLL-rearranged pro-B and CD10- pre-B ALL cases had similar clinical and immunophenotypic (coexpression of CDw65 and CD15) features at initial diagnosis. CONCLUSIONS: The striking similarities between the two CD10 ALL subsets imply that CD10- pre-B ALL variants may represent pro-B ALL cases that maintained the propensity to rearrange and express their immunoglobulin heavy chain rather than actual pre-B ALL forms transformed at this later stage of B-cell differentiation. However, direct experimental data are needed to confirm this observation. PMID- 16707594 TI - CpG island methylation status in gastric carcinoma with and without infection of Epstein-Barr virus. AB - PURPOSE: EBV-associated gastric carcinoma shows global CpG island methylation of the promoter region of various cancer-related genes. To further clarify the significance of CpG island methylator phenotype (CIMP) status in gastric carcinoma, we investigated methylation profile and clinicopathologic features including overall survival in four subgroups defined by EBV infection and CIMP status: EBV-associated gastric carcinoma and EBV-negative/CIMP-high (H), EBV intermediate (I), and EBV-negative (N) gastric carcinoma. EXPERIMENTAL DESIGN: Methylation-specific PCR was applied to 106 gastric carcinoma cases. CIMP-N, CIMP I, and CIMP-H status was determined by the number (0, 1-3, and 4-5, respectively) of methylated marker genes (LOX, HRASLS, FLNc, HAND1, and TM), that were newly identified as highly methylated in gastric cancer cell lines. The methylation status of 10 other cancer-related genes (p14, p15, p16, p73, TIMP-3, E-cadherin, DAPK, GSTP1, hMLH1, and MGMT) was also evaluated. RESULTS: Nearly all (14 of 15) of EBV-associated gastric carcinoma exhibited CIMP-H, constituting a homogenous group (14%). EBV-negative gastric carcinoma consisted of CIMP-H (24%), CIMP-I (38%), and CIMP-N (24%). EBV-associated gastric carcinoma showed significantly higher frequencies of methylation of cancer-related genes (mean number +/- SD = 6.9 +/- 1.5) even if compared with EBV-negative/CIMP-H gastric carcinoma (3.5 +/- 1.8). Among EBV-negative gastric carcinoma subgroups, CIMP-H gastric carcinoma showed comparatively higher frequency of methylation than CIMP-I or CIMP-N, especially of p16 and hMLH1. CIMP-N gastric carcinoma predominantly consisted of advanced carcinoma with significantly higher frequency of lymph node metastasis. The prognosis of the patients of CIMP-N was significantly worse compared with other groups overall by univariate analysis (P = 0.0313). CONCLUSION: The methylation profile of five representative genes is useful to stratify gastric carcinomas into biologically different subgroups. EBV-associated gastric carcinoma showed global CpG island methylation, comprising a pathogenetically distinct subgroup in CIMP-H gastric carcinoma. PMID- 16707595 TI - Glucocorticoids suppress tumor angiogenesis and in vivo growth of prostate cancer cells. AB - PURPOSE: Glucocorticoids, such as prednisone, hydrocortisone, and dexamethasone, are known to produce some clinical benefit for patients with hormone-refractory prostate cancer (HRPC). However, the underlying mechanisms by which glucocorticoids affect HRPC growth are not well established as yet. Here, we hypothesize that the therapeutic effect of glucocorticoids on HRPC can be attributed to a direct inhibition of angiogenesis through the glucocorticoid receptor by down-regulating two major angiogenic factors, vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8). EXPERIMENTAL DESIGN: The effects of dexamethasone on VEGF and IL-8 expression and cell proliferation were examined using DU145, which expresses glucocorticoid receptor. The effects of dexamethasone on DU145 xenografts were determined by analyzing VEGF and IL-8 gene expression, microvessel density, and tumor volume. RESULTS: Dexamethasone significantly down-regulated VEGF and IL-8 gene expression by 50% (P < 0.001) and 89% (P < 0.001), respectively, and decreased VEGF and IL-8 protein production by 55% (P < 0.001) and 74% (P < 0.001), respectively, under normoxic condition. Similarly, hydrocortisone down-regulated VEGF and IL-8 gene expression. The effects of dexamethasone were completely reversed by the glucocorticoid receptor antagonist RU486. Even under hypoxia-like conditions, dexamethasone inhibited VEGF and IL-8 expression. In DU145 xenografts, dexamethasone significantly decreased tumor volume and microvessel density and down-regulated VEGF and IL-8 gene expression, whereas dexamethasone did not affect the in vitro proliferation of the cells. CONCLUSION: Glucocorticoids suppressed androgen-independent prostate cancer growth possibly due to the inhibition of tumor-associated angiogenesis by decreasing VEGF and IL-8 production directly through glucocorticoid receptor in vivo. PMID- 16707596 TI - 17Beta-estradiol induces down-regulation of Cap43/NDRG1/Drg-1, a putative differentiation-related and metastasis suppressor gene, in human breast cancer cells. AB - PURPOSE: Cap43 is known as a nickel- and calcium-inducible gene. In the present study, we examined whether 17beta-estradiol (E2) could affect the expression of Cap43 in breast cancer. EXPERIMENTAL DESIGN: Real-time PCR, immunoblotting, and immunocytochemistry were used to examine the expression of Cap43 and estrogen receptor-alpha (ER-alpha) in breast cancer cell lines. MDA-MB-231 and SK-BR-3 cell lines were transfected with ER-alpha cDNA to establish cells overexpressing ER-alpha. Immunohistochemistry was used to evaluate the expression of the Cap43 protein in breast cancer patients (n = 96), and the relationship between Cap43 expression and clinicopathologic findings was examined. RESULTS: Of the eight cell lines, four expressed higher levels of Cap43 with very low levels of ER alpha, whereas the other four expressed lower levels of Cap43 with high ER-alpha levels. Treatment with E2 decreased the expression of Cap43 dose-dependently in ER-alpha-positive cell lines but not in ER-alpha-negative lines. Administration of antiestrogens, tamoxifen and ICI 182780, abrogated the E2-induced down regulation of Cap43. Overexpression of ER-alpha in both ER-alpha-negative cell lines, SK-BR-3 and MDA-MB-231, resulted in down-regulation of Cap43. Immunostaining studies showed a significant correlation between Cap43 expression and the histologic grade of tumors (P = 0.0387). Furthermore, Cap43 expression was inversely correlated with the expression of ER-alpha (P = 0.0374). CONCLUSIONS: E2-induced down-regulation of Cap43 seems to be mediated through ER alpha-dependent pathways in breast cancer cells both in culture and in patients. Cap43 has potential as a molecular marker to determine the therapeutic efficacy of antiestrogenic anticancer agents in breast cancer. PMID- 16707597 TI - Phosphatidylinositol 3'-kinase/AKT signaling is activated in medulloblastoma cell proliferation and is associated with reduced expression of PTEN. AB - PURPOSE: Medulloblastomas represent the most frequent malignant brain tumors of childhood. They are supposed to originate from cerebellar neural precursor cells. Recently, it has been shown that Sonic Hedgehog-induced formation of medulloblastoma in an animal model is significantly enhanced by activation of the phosphatidylinositol 3'-kinase (PI3K) signaling pathway. EXPERIMENTAL DESIGN: To examine a role for PI3K/AKT signaling in the molecular pathogenesis of human medulloblastoma, we did an immunohistochemical study of the expression of Ser473 phosphorylated (p)-AKT protein in 22 medulloblastoma samples: All samples displayed p-AKT expression. To investigate if an activated PI3K/AKT pathway is required for medulloblastoma cell growth, we treated five human medulloblastoma cell lines with increasing concentrations of the PI3K inhibitor LY294002 and analyzed cellular proliferation and apoptosis. The antiproliferative effect could be antagonized by overexpressing constitutively active AKT. As the activation of PI3K/AKT signaling may be associated with alterations of the PTEN gene located at 10q23.3, a chromosomal region subject to frequent allelic losses in medulloblastoma, we screened PTEN for mutations and mRNA expression. RESULTS: Proliferation of all of the medulloblastoma cell lines was dependent on PI3K/AKT signaling, whereas apoptosis was not prominently affected. Allelic loss was detected in 16% of the cases. One medulloblastoma cell line was found to carry a truncating mutation in the PTEN coding sequence. Even more important, PTEN mRNA and protein levels were found to be significantly lower in medulloblastomas compared with normal cerebellar tissue of different developmental stages. Reduction of PTEN expression was found to be associated with PTEN promoter hypermethylation in 50% of the tumor samples. CONCLUSIONS: We conclude that activation of the PI3K/AKT pathway constitutes an important step in the molecular pathogenesis of medulloblastoma and that dysregulation of PTEN may play a significant role in this context. PMID- 16707598 TI - Peroxisome proliferator-activated receptor alpha activation decreases metastatic potential of melanoma cells in vitro via down-regulation of Akt. AB - PURPOSE: Peroxisome proliferator-activated receptors (PPAR) regulate lipid and glucose metabolism but their anticancer properties have been recently studied as well. We previously reported the antimetastatic activity of the PPARalpha ligand, fenofibrate, against melanoma tumors in vivo. Here we investigated possible molecular mechanisms of fenofibrate anti metastatic action. EXPERIMENTAL DESIGN: Monolayer cultures of mouse (B16F10) and human (SkMell88) melanoma cell lines, soft agar assay, and cell migration assay were used in this study. In addition, we analyzed PPARalpha expression and its transcriptional activity in response to fenotibrate by using Western blots and liciferase-based reporter system. RESULTS: Fenofibrate inhibited migration of B16F10 and SkMel188 cells in Transwell chambers and colony formation in soft agar. These effects were reversed by PPAR inhibitor, GW9662. Western blot analysis revealed time-dependent down-regulation of Akt and extracellular signal-regulated kinase l/2 phosphorylation in fenofibrate-treated cells. A B16F10 cell line stably expressing constitutively active Akt mutant was resistant to fenofibrate. In contrast, Akt gene silencing with siRNA mimicked the fenofibrate action and reduced the migratory ability of B16F1O cells. In addition, fenofibrate strongly sensitized BI6FIO cells to the proapoptotic drug staurosporine, further supporting the possibility that fenofibrate-induced down-regulation of Akt function contributes to fenofibrate mediated inhibition of metastatic potential in this experimental model. CONCLUSIONS: Our results show that the PPAR-dependent antimetastatic activity of fenofibrate involves down-regulation of Akt phosphorylation and suggest that supplementation with this drug may improve the effectiveness of melanoma chemotherapy. PMID- 16707599 TI - Achieving a major molecular response at the time of a complete cytogenetic response (CCgR) predicts a better duration of CCgR in imatinib-treated chronic myeloid leukemia patients. AB - PURPOSE: Most patients with chronic-phase chronic myeloid leukemia (CML) who receive imatinib achieve a complete cytogenetic remission (CCgR) and low levels of BCR-ABL transcripts. CCgR is durable in the majority of patients but relapse occurs in a subset. EXPERIMENTAL DESIGN: To determine the potential of quantitative reverse transcription-PCR of BCR-ABL to predict cytogenetic relapse, we serially monitored residual disease in 97 CML patients with an imatinib induced CCgR. Patients with late chronic phase CML after IFN-alpha failure were treated with imatinib (400 mg daily). RESULTS: During the imatinib median follow up time of 36 months (range, 12-54 months), disease monitoring occurred by cytogenetics and quantitative PCR. Twenty percent of patients experienced cytogenetic relapse at a median of 18 months after CCgR and a median of 24 months after starting imatinib. None of the possible prognostic factors studied in univariate and multivariate analyses seemed to predict for loss of cytogenetic response but the reduction of BCR-ABL transcript levels at the time of CCgR is an important prognostic factor. CONCLUSIONS: In our study, we showed not only that achieving a major molecular remission at 12 months is predictive of a durable cytogenetic remission but also that patients who achieved a major molecular remission (expressed both as the BCR-ABL/beta2 microglobulin ratio % <0.0005 and as a 3-log reduction from median baseline value) already at the time of first achieving a CCgR have significantly longer cytogenetic remission durations than those without this magnitude of molecular response (P < 0.05). PMID- 16707600 TI - NOTCH1 mutations in T-cell acute lymphoblastic leukemia: prognostic significance and implication in multifactorial leukemogenesis. AB - PURPOSE: NOTCH signaling pathway is essential in T-cell development and NOTCH1 mutations are frequently present in T-cell acute lymphoblastic leukemia (T-ALL). To gain insight into its clinical significance, NOTCH1 mutation was investigated in 77 patients with T-ALL. EXPERIMENTAL DESIGN: Detection of NOTCH1 mutation was done using reverse transcription-PCR amplification and direct sequencing, and thereby compared according to the clinical/biological data of the patients. RESULTS: Thirty-two mutations were identified in 29 patients (with dual mutations in 3 cases), involving not only the heterodimerization and proline/glutamic acid/serine/threonine domains as previously reported but also the transcription activation and ankyrin repeat domains revealed for the first time. These mutations were significantly associated with elevated WBC count at diagnosis and independently linked to short survival time. Interestingly, the statistically significant difference of survival according to NOTCH1 mutations was only observed in adult patients (>18 years) but not in pediatric patients (< or = 18 years), possibly due to the relatively good overall response of childhood T-ALL to the current chemotherapy. NOTCH1 mutations could coexist with HOX11, HOX11L2, or SIL-TAL1 expression. The negative effect of NOTCH1 mutation on prognosis was potentiated by HOX11L2 but was attenuated by HOX11. CONCLUSION: NOTCH1 mutation is an important prognostic marker in T-ALL and its predictive value could be even further increased if coevaluated with other T-cell-related regulatory genes. NOTCH pathway thus acts combinatorially with oncogenic transcriptional factors on T-ALL pathogenesis. PMID- 16707601 TI - Glutathione S-transferase P1 polymorphism (Ile105Val) predicts cumulative neuropathy in patients receiving oxaliplatin-based chemotherapy. AB - PURPOSE: Glutathione S-transferases (GST) are xenobiotic metabolizing enzymes involved in the detoxification of a variety of chemotherapeutic drugs, including platinum derivatives. Genetic polymorphisms of GSTs have been associated with enzyme activity variations. Thus, a study was done to investigate the relationship between GST polymorphisms and oxaliplatin-related cumulative neuropathy in gastrointestinal cancer patients treated with oxaliplatin-based chemotherapy. EXPERIMENTAL DESIGN: Ninety patients were included. Clinical neurologic evaluation was done at baseline and before each cycle of treatment. We determined genetic variants for GSTP1 exon 5 (Ile105Val), GSTP1 exon 6 (Ala114Val), GSTM1 (homozygous deletion), and GSTT1 (homozygous deletion). We conducted analyses in a subgroup of 64 patients receiving a minimal cumulative dose of 500 mg/m2 of oxaliplatin to examine whether the GST polymorphisms are associated with oxaliplatin-related cumulative neuropathy. RESULTS: Among patients receiving a minimal cumulative dose of 500 mg/m2 of oxaliplatin, 15 patients showed clinically evident oxaliplatin-related cumulative neuropathy scored grade 3 according to an oxaliplatin-specific scale. The oxaliplatin related cumulative neuropathy scored grade 3 was significantly more frequent in patients homozygous for the GSTP1 105Ile allele than in patients homozygous or heterozygous for the GSTP1 105Val allele (odds ratio, 5.75; 95% confidence interval, 1.08-30.74; P = 0.02). No association was found with respect to any of the GSTM1, GSTT1, or GSTP1 exon 6 genotypes. CONCLUSIONS: The results of the current study suggest that the 105Val allele variant of the GSTP1 gene at exon 5 confers a significantly decreased risk of developing severe oxaliplatin-related cumulative neuropathy. PMID- 16707602 TI - Clinical significance of TROP2 expression in colorectal cancer. AB - PURPOSE AND EXPERIMENTAL DESIGN: To identify cancer-related genes, the expression profiles of colorectal cancer cells and normal epithelial cells were examined and compared using laser microdissection and cDNA microarray analysis. From these combined techniques, several cancer-related genes, including TROP2, were identified. TROP2 is known as a calcium signal transducer and is highly expressed in several types of tumors. However, no studies have investigated the significance of TROP2 expression in colorectal cancer. Thus, the expression status of TROP2 was investigated in 74 colorectal cancer samples by quantitative real-time reverse transcription-PCR and immunohistochemical studies. RESULTS: Laser microdissection and cDNA microarray analysis showed that there were 84 overexpressed genes in cancer cells. One of the highly overexpressed genes was TROP2. Quantitative real-time reverse transcription-PCR showed that TROP2 expression in cancer samples was significantly higher than in normal samples (P < 0.001). The samples were divided into high (n = 26) and low (n = 48) TROP2 expression groups. The cases with high TROP2 expression showed a higher frequency of liver metastasis (P = 0.005) and more cancer-related death (P = 0.046). Those cases also had an inclination of deeper depth of invasion (P = 0.064) and more lymph node metastasis (P = 0.125). Interestingly, the patients with high TROP2 expression tumors had poorer prognosis (P = 0.0036). Multivariate analysis showed that TROP2 expression status was an independent prognostic factor (relative risk, 2.38; 95% confidence interval, 1.29-4.74; P < 0.01). CONCLUSION: TROP2 is one of the cancer-related genes that correlates with biological aggressiveness and poor prognosis of colorectal cancer. Thus, TROP2 is a possible candidate gene for diagnosis and molecular target therapy of colorectal cancer. PMID- 16707603 TI - Clinical and immunologic responses to active specific cancer vaccines in human colorectal cancer. AB - Colorectal cancer is a common malignant disease, which, despite some progress, still requires improved therapeutic options. Several clinical studies have used active specific immunotherapy (i.e., vaccination) in colorectal cancer. However, the literature still lacks a comprehensive meta-analysis of this approach in advanced colorectal cancer. We did a systematic review with a meta-analysis of clinical studies to evaluate the objective clinical and immunologic response to active specific immunotherapy in patients with colorectal cancer. We conducted a search of Medline and the Web of Science, manually reviewed the literature, and consulted with experts. Criteria for including studies were colorectal cancer patients, active specific immunotherapy to induce a response directed against cancer or cancer antigens, an evaluable tumor burden (i.e., advanced or metastatic colorectal cancer), and precise classification of the patient, disease, and response. Response rates were assessed according to WHO criteria. Primary end points were the objective clinical response rate and the rate of immunologic responses. The secondary end point was the distribution of immune and clinical responses in relation to the route of vaccination and the type of vaccine. Thirty-two phase I/II studies reporting on 527 patients with advanced or metastatic colorectal cancer met all inclusion criteria. Pooled analysis showed an overall response rate (complete response + partial response) of 0.9% for advanced/metastatic colorectal cancer patients who underwent active specific immunization with a broad variety of substances (e.g., autologous tumor cells, peptide vaccine, dendritic cells, idiotypic antibody, and virus-based vaccine). Humoral immune responses were reported in 59%, and cellular ones were reported in 44% of the cases. Mixed or minor responses and disease stabilization are described in 1.9% and 8.3% of colorectal cancer patients, respectively. Pooled results of clinical trials reveal a very weak clinical response rate of <1% for active specific immunization procedures currently available for advanced colorectal cancer. Immune response induction is described in approximately half the patients. PMID- 16707604 TI - Prognostic significance of activated AKT expression in soft-tissue sarcoma. AB - PURPOSE: AKT is a serine/threonine kinase which is important in tumorigenesis. Several molecules involved in AKT pathway are dysregulated in various kinds of human cancers. PATIENTS AND METHODS: Ninety-three patients (53 males and 40 females), ages ranging from 19 to 77 years (median, 57 years), with localized soft-tissue sarcomas arising in the trunk and extremities, were analyzed. Immunoperoxidase procedure (avidin-biotin complex method) was done on paraffin embedded sections with anti-phosphorylated AKT (Thr308), anti-phosphorylated p44/42 extracellular signal-regulated kinase 1 and 2 (ERK1/2) (Thr202/Tyr204), anti-phosphorylated forkhead in rhabdomyosarcoma (FKHR) (Ser256), and anti-Ki 67 antibodies. Expression levels of phosphorylated AKT (p-AKT), phosphorylated ERK1/2 (p-ERK1/2), and phosphorylated FKHR (p-FKHR) were categorized as either weaker (level 1) or equal to or stronger (level 2) compared with those in the endothelial cells of the same specimens. Percentage of cells showing intranuclear staining with Ki-67 was shown as the Ki-67 labeling index (LI). Cases were divided into two groups: level 1, Ki-67 LI < 20%; level 2, Ki-67 LI > or = 20%. RESULTS: Twenty-six (28.0%), 6 (6.5%), and 46 (44.1%) of the tumors showed level 2 expression for p-AKT, p-ERK1/2, and Ki-67 LI, respectively. Tumors with level 2 p-AKT expression showed a higher ratio of level 2 p-FKHR expression (P < 0.01). Multivariate analysis revealed p-AKT expression and Ki-67 LI to be independent prognosticators for overall survival, and p-AKT expression for disease-free survival. CONCLUSION: p-AKT expression level is a significant prognosticator in soft-tissue sarcoma. PMID- 16707605 TI - Epidermal growth factor receptor messenger RNA expression, gene dosage, and gefitinib sensitivity in non-small cell lung cancer. AB - PURPOSE: Epidermal growth factor receptor (EGFR) mRNA expression and EGFR gene dosage by quantitative PCR in tumor samples obtained from patients with gefitinib treated non-small cell lung cancer were analyzed in order to determine the association with treatment outcome, clinical, and biological features [EGFR copy number by fluorescent in situ hybridization (FISH), EGFR tyrosine kinase mutations, and EGFR protein expression]. EXPERIMENTAL DESIGN: EGFR mRNA expression was measured by real-time quantitative reverse transcription-PCR in 64 patients, and EGFR gene dosage was analyzed by real-time quantitative PCR in 82 patients from paraffin-embedded specimens. RESULTS: EGFR mRNA expression was higher in responders to gefitinib as compared with nonresponders (P = 0.012). Patients with high EGFR mRNA expression (>5.01) had 43% response probability, whereas patients with low EGFR mRNA expression had 8% response probability (P = 0.006). Patients with high EGFR mRNA expression had longer median progression free (5.3 versus 2.8 months, P = 0.028) but not overall survival (13.8 versus 10.9 months, P = 0.87). EGFR mRNA expression was higher in FISH-positive patients (P = 0.001) and in patients with positive EGFR immunostaining (P < 0.001) but not in patients with EGFR mutations (P = 0.19). EGFR gene dosage did not predict response (P = 0.54), progression-free (P = 0.73), or overall survival (P = 0.89). EGFR gene dosage was not associated with FISH positivity (P = 0.15), relative mRNA expression (P = 0.27), EGFR mutation status (P = 0.39), and EGFR protein expression (P = 0.35). CONCLUSION: EGFR mRNA expression is a predictive biomarker for response to gefitinib and to progression-free survival after gefitinib treatment. EGFR gene dosage is neither predictive for response nor progression free nor overall survival. PMID- 16707606 TI - Phase I trial of the trifunctional anti-HER2 x anti-CD3 antibody ertumaxomab in metastatic breast cancer. AB - PURPOSE: Ertumaxomab is an intact bispecific antibody targeting HER2/neu and CD3 with selective binding to activatory Fcgamma type I/III receptors, resulting in the formation of a tri-cell complex between tumor cells, T cells, and accessory cells. Patients with metastatic breast cancer were enrolled into a multicenter phase I dose-escalating trial. EXPERIMENTAL DESIGN: Three ascending doses of ertumaxomab (10-200 microg) were administered i.v. on day 1, 7 +/- 1, and 13 +/- 1. Safety and tolerability were the primary objectives. Secondary objectives were antitumor activity and different immunologic variables. RESULTS: Fifteen out of 17 enrolled patients completed the study. One hundred micrograms was identified as the maximal tolerable single dose. Most drug-related adverse events were mild and transient including fever (94%), rigors (47%), headache (35%), nausea (29%), vomiting (29%). Grades 3 and 4 (Common Toxicity Criteria) were lymphocytopenia (76%) and elevation of liver enzymes (47%). One patient (200 mug dose) developed severe hypotension and respiratory distress syndrome, another patient (150 mug dose) developed a systemic inflammatory response syndrome and acute renal failure. Aggravation of congestive heart failure was seen in one patient with preexisting ventricular dysfunction after administration of the third dose (200 microg). All adverse events were fully reversible. Antitumor response was seen in 5 out of 15 evaluable patients (one with a complete response, two with partial responses, two with stable disease) at dose levels of > or = 100 microg. Measurements of cytokines (interleukin-6, interleukin-2, tumor necrosis factor alpha, and IFN-gamma) suggest a strong T helper cell type 1-associated immune response. The induction of human anti-mouse/anti-rat antibodies was detected in 5 out of 16 (31%) patients. DISCUSSION: Treatment with triple infusions of ertumaxomab yields a strong immunologic response. Doses up to 100 microg can be safely infused with close monitoring of patients. The observed clinical responses are encouraging and indicate antitumor efficacy. PMID- 16707607 TI - Phase II clinical trial results involving treatment with low-dose daily oral cyclophosphamide, weekly vinblastine, and rofecoxib in patients with advanced solid tumors. AB - PURPOSE: Preclinical studies indicate that conventional chemotherapeutic agents given continuously at low doses (metronomic chemotherapy) may provide an improved therapeutic index. Cyclophosphamide and vinblastine have been best studied in experimental models, where tumor growth inhibition is achieved, at least in part, through antiangiogenic mechanisms. EXPERIMENTAL DESIGN: Fifty patients with advanced solid tumors were enrolled in this phase II trial, 43 of whom had received at least one prior chemotherapy regimen. Patients were required to have Eastern Cooperative Oncology Group performance status of < or = 2, a life expectancy of >3 months, and at least one measurable lesion. All patients received oral cyclophosphamide (50 mg) and rofecoxib (25 mg) daily in addition to weekly injections of vinblastine (3 mg/m2). Half of the patients also received minocycline (100 mg) orally twice daily with the intent of further inhibiting tumor angiogenesis. The primary end point of the study was clinical benefit, defined as the percentage of patients experiencing an objective response or exhibiting stable disease for at least 6 months. RESULTS: For the 47 eligible patients, there were two (4%) complete responses and four (9%) partial responses, for an overall objective response rate of 13%. An additional eight patients achieved disease stabilization (stable disease > or = 6 months) (17%). The primary end point of clinical benefit was therefore 30%, (95% confidence interval, 16-44%). The median progression-free survival for all patients was 103 days and 289 days for patients experiencing clinical benefit. The incidence of patients experiencing grade 3/4 toxicities were as follows: neutropenia (10/2), anemia (2/0), and thrombocytopenia (1/0). No patients developed grade 3 or 4 nausea, vomiting, mucositis, or alopecia. CONCLUSIONS: This low-dose regimen consisting of daily oral cyclophosphamide and weekly vinblastine injections given concurrently with rofecoxib is associated with minimal toxicity and provides significant clinical benefit to patients with advanced solid tumors. These results are particularly encouraging given the nature of the study population and indicate that this approach merits further investigation in specific disease site studies. PMID- 16707608 TI - Effect of cA2 anti-tumor necrosis factor-alpha antibody therapy on hematopoiesis of patients with myelodysplastic syndromes. AB - PURPOSE: Tumor necrosis factor alpha (TNF-alpha) plays a prominent role in the pathophysiology of myelodysplastic syndromes (MDS). The aim of this study was to explore the biological and immunoregulatory effect of the treatment with the anti tumor necrosis factor-alpha monoclonal antibody cA2 on bone marrow (BM) progenitor/precursor and stromal cells and lymphocyte subsets, as well as the clinical response in MDS patients. EXPERIMENTAL DESIGN: Ten low-intermediate risk MDS patients received i.v. cA2 (3 mg/kg) at weeks 0, 2, 6, and 12. The number, survival, and clonogenic potential of BM progenitor/precursor cells, the hematopoiesis-supporting capacity of BM stromal cells, and the lymphocyte activation status were investigated in the patients at baseline and following treatment using flow cytometry, clonogenic assays, and long-term BM cultures (LTBMC). Clinical response was evaluated according to standardized criteria. RESULTS: cA2 administration reduced the proportion of apoptotic and Fas+ cells in the CD34+ cell compartment (P = 0.0215 and P = 0.0344, respectively) and increased the clonogenic potential of BM mononuclear and CD34+ cells (P = 0.0399 and P = 0.0304, respectively) compared with baseline. The antibody reduced tumor necrosis factor-alpha levels in LTBMC supernatants (P = 0.0043) and significantly improved the hematopoiesis-supporting capacity of LTBMC adherent cells. The proportion of activated peripheral blood and BM T-lymphocytes decreased significantly after treatment, suggesting an immunomodulatory effect of cA2. Two patients displayed minor hematologic responses whereas the remaining patients displayed stable disease with no disease progression. CONCLUSIONS: The encouraging biological insights from cA2 administration may be useful in conducting further clinical trials using cA2 for selected MDS patients, particularly those with evidence of immune-mediated inhibition of hematopoiesis. PMID- 16707609 TI - Fenretinide activity in retinoid-resistant oral leukoplakia. AB - PURPOSE: To test the hypothesis that the retinamide N-(4-hydroxyphenyl)retinamide (fenretinide) would be clinically active potentially via receptor-independent apoptosis and receptor-dependent effects in natural retinoid-resistant oral leukoplakia patients--the first test of this hypothesis in any in vivo setting. EXPERIMENTAL DESIGN: A phase II trial of fenretinide (200 mg/d for 3 months) in oral leukoplakia patients who had not responded (de novo resistance) or who had responded and then relapsed (acquired resistance) to previous treatment with natural retinoids. We analyzed apoptosis via the terminal deoxynucleotidyl transferase-mediated nick end labeling in situ DNA fragmentation assay. RESULTS: We accrued 35 evaluable patients with retinoid-resistant oral leukoplakia, 12 (34.3%) had partial responses to fenretinide (95% confidence interval, 19.2 52.4%), and response was associated with acquired resistance to natural retinoids (P = 0.015, Fisher's exact test). Nine responders progressed within 9 months of stopping fenretinide. Toxicity was minimal and compliance was excellent. Mean apoptosis values (SE) increased from 0.35% (0.25%) at baseline to 1.18% (0.64%) at 3 months (P = 0.001, sign test); this increase did not correlate with clinical response. The increases in 3-month mean serum concentrations of fenretinide (0.23 micromol/L) and N-(4-methoxyphenyl)retinamide (0.57 micromol/L) correlated with decreased retinol concentrations [Spearman correlation coefficient of -0.57 (P = 0.001) and -0.43 (P = 0.01), respectively]. CONCLUSIONS: Low-dose fenretinide was clinically active and produced a small increase in apoptosis in retinoid resistant oral leukoplakia. PMID- 16707610 TI - Carbogen and nicotinamide increase blood flow and 5-fluorouracil delivery but not 5-fluorouracil retention in colorectal cancer metastases in patients. AB - PURPOSE: To examine whether carbogen and nicotinamide increases 5-fluorouracil (5 FU) delivery to colorectal cancer metastases. EXPERIMENTAL DESIGN: Six patients were scanned using positron emission tomography. Two scans were done to coincide with the start of separate chemotherapy cycles. At the second positron emission tomography session, 60 mg/kg nicotinamide was given orally 2 to 3 hours before 10 minute carbogen inhalation. In the middle of carbogen treatment, [15O]H2O (to measure regional tissue perfusion) and then [18F]5-FU (to measure 5-FU tissue pharmacokinetics) were administered. RESULTS: Regions of interest were drawn in 12 liver metastases, 6 spleens, 6 livers, and 12 kidneys. Nicotinamide and carbogen administration increased mean blood pO2 from 93 mm Hg (95% confidence interval, 79-198) to 278 mm Hg (95% confidence interval, 241-316; P = 0.031). Regional perfusion (mL(blood)/min/mL(tissue)) increased in metastases (mean change = 52%, range -32% to +261%, P = 0.024), but decreased in kidney (mean change = -42%, range -82% to -11%, P = 0.0005) and liver (mean change = -34%, range -43% to -26%, P = 0.031). 5-FU uptake at 3.75 minutes (m(2)/mL) increased in tumor (mean change = 40%, range -39% to +196%, P = 0.06) and decreased in kidney (mean change = -25%, range -71% to 12%, P = 0.043). 5-FU delivery measured as K1 increased in tumor (mean change = 74%, range -23% to +293%, P = 0.0039). No differences were seen in [18F]5-FU tumor exposure (net area under curve) and retention. CONCLUSION: Nicotinamide and carbogen administration can increase 5-FU delivery to colorectal cancer liver metastases. Despite an increase in perfusion and 5-FU delivery, the effects were not directly related and did not increase 5 FU retention or tissue exposure. PMID- 16707611 TI - Phase II trial of bevacizumab in combination with weekly docetaxel in metastatic breast cancer patients. AB - PURPOSE: To evaluate the safety and efficacy of bevacizumab and weekly docetaxel as first- or second-line therapy in patients with metastatic breast cancer (MBC). PATIENTS AND METHODS: Twenty-seven MBC patients received i.v. bevacizumab at 10 mg/kg on days 1 and 15 in combination with i.v. docetaxel 35 mg/m2 on days 1, 8, and 15 of a 28-day cycle. Primary end points were to assess toxicity, overall response rate, and progression-free survival. A secondary end point was to assess the relationship between plasma endothelial and cell adhesion markers and clinical outcomes. RESULTS: One-hundred fifty-eight treatment cycles were administered with a median of six cycles (range 1-15 cycles) per patient. The most common grade 4 toxicities per patient were as follows: 2 (7%)-pulmonary embolus, 1 (4%)-febrile neutropenia, and 1 (4%)-infection; grade 3 toxicities were 4 (15%)-neutropenia, 4 (15%)-fatigue, 2 (7%)-neuropathy, 2 (7%)-athralgias, 2 (7%)-stomatitis, 1 (7%)-pleural effusion, and 1 (4%)-hypertension. The overall response rate was 52% [95% confidence interval (95% CI), 32-71%], median response duration was 6.0 months (95% CI, 4.6-6.5 months), and the median progression-free survival was 7.5 months (95% CI, 6.2-8.3 months). In hypothesis-generating univariate and limited multivariate analyses, E-selectin was statistically significantly associated with response to the combination. CONCLUSION: Bevazicumab in combination with weekly docetaxel is active with acceptable toxicities in MBC. Additional studies evaluating E-selectin as a marker of response to bevacizumab-containing chemotherapy are warranted. PMID- 16707612 TI - Detection and functional analysis of CD8+ T cells specific for PRAME: a target for T-cell therapy. AB - PURPOSE: Preferentially expressed antigen on melanomas (PRAME) is an interesting antigen for T-cell therapy because it is frequently expressed in melanomas (95%) and other tumor types. Moreover, due to its role in oncogenic transformation, PRAME-negative tumor cells are not expected to easily arise and escape from T cell immunity. The purpose of this study is to investigate the usefulness of PRAME as target for anticancer T-cell therapies. EXPERIMENTAL DESIGN: HLA-A*0201 subtyped healthy individuals and advanced melanoma patients were screened for CD8+ T cells directed against previously identified HLA-A*0201-binding PRAME peptides by IFN-gamma enzyme-linked immunosorbent spot assays and tetramer staining. PRAME-specific T-cell clones were isolated and tested for recognition of melanoma and acute lymphoid leukemia (ALL) cell lines. PRAME mRNA expression was determined by quantitative real-time reverse transcription-PCR. RESULTS: In 30% to 40% of healthy individuals and patients, PRA(100-108)-specific CD8+ T cells were detected both after in vitro stimulation and directly ex vivo after isolation by magnetic microbeads. Although CD45RA- memory PRA(100-108)-specific T cells were found in some individuals, the majority of PRA(100-108)-tetramer+ T cells expressed CD45RA, suggesting a naive phenotype. PRA(100-108)-tetramer+ T cell clones were shown to recognize and lyse HLA-A*0201+ and PRAME+ melanoma but not ALL cell lines. Quantitative real-time reverse transcription-PCR showed significantly lower PRAME mRNA levels in ALL than in melanoma cell lines, suggesting that PRAME expression in ALL is below the recognition threshold of our PRA(100-108)-tetramer+ T cells. CONCLUSION: These data support the usefulness of PRAME and in particular the PRA(100-108) epitope as target for T-cell therapy of PRAME-overexpressing cancers. PMID- 16707613 TI - Infectivity-enhanced adenoviruses deliver efficacy in clinical samples and orthotopic models of disseminated gastric cancer. AB - PURPOSE: Metastatic gastric cancer remains a common and devastating disease without curative treatment. Recent proof-of-concept clinical trials have validated gene therapy with adenoviruses as an effective and safe modality for the treatment of cancer. However, expression of the primary coxsackie-adenovirus receptor is variable in advanced cancers, and therefore, the use of heterologous receptors could be advantageous. EXPERIMENTAL DESIGN: Here, we used capsid modified adenoviruses for increasing the transduction and subsequent antitumor efficacy. 5/3 chimeric viruses have a serotype 3 knob which allows binding to a receptor distinct from coxsackie-adenovirus receptor. The fiber of Ad5lucRGD is modified with an integrin-targeted motif. Polylysine motifs, pK7 and pK21, bind to heparan sulfates. Oncolytic adenoviruses replicate in and kill tumor cells selectively. Gastric cancer cell lines and fresh clinical samples from patients were infected with transductionally targeted viruses. Capsid-modified oncolytic adenoviruses were used in cell killing experiments. To test viral transduction and therapeutic efficacy in vivo, we developed orthotopic mouse models featuring i.p. disseminated human gastric cancer, which allowed the evaluation of biodistribution and antitumor efficacy in a system similar to humans. RESULTS: Capsid modifications benefited gene transfer efficiency and cell killing in gastric cancer cell lines and clinical samples in vitro and in vivo. Modified oncolytic adenoviruses significantly increased the survival of mice with orthotopic gastric cancer. CONCLUSIONS: These preclinical data set the stage for the clinical evaluation of safety and efficacy in patients with disease refractory to current modalities. PMID- 16707615 TI - Effective treatment of pancreatic cancer xenografts with a conditionally replicating virus derived from type 2 herpes simplex virus. AB - PURPOSE: Pancreatic cancer is a devastating disease that is almost universally fatal because of the lack of effective treatments. We recently constructed a novel oncolytic virus (FusOn-H2) from the type 2 herpes simplex virus. Because the replication potential of FusOn-H2 depends on the activation of the Ras signaling pathway, we evaluated its antitumor effect against pancreatic cancer, which often harbors K-ras gene mutations. EXPERIMENTAL DESIGN: Human pancreatic cancer xenografts were established in nude mice either s.c. or orthotopically (n = 8/group). FusOn-H2 was injected either directly (s.c. tumors) or by the i.v. or i.p. route (orthotopic tumors). Tumor volume, weight, and survival time were recorded for each animal. Statistical analyses were done by Student's t test. RESULTS: A single intratumor injection of FusOn-H2 completely eradicated s.c. pancreatic cancers in all animals. Systemic injection of the oncolytic virus produced clear antitumor effects but did not abolish tumors in any animal. The most striking antitumor effect was seen when the virus was given i.p. Delivery of FusOn-H2 by this route completely eradicated established orthotopic tumors in 75% of the animals and completely prevented local metastases. CONCLUSIONS: FusOn-H2 has potent activity against human pancreatic cancer xenografts and may be a promising candidate for investigative virotherapy of this malignancy. PMID- 16707614 TI - Real-time, image-guided, convection-enhanced delivery of interleukin 13 bound to pseudomonas exotoxin. AB - PURPOSE: To determine if the tumor-targeted cytotoxin interleukin 13 bound to Pseudomonas exotoxin (IL13-PE) could be delivered to the brainstem safely at therapeutic doses while monitoring its distribution in real-time using a surrogate magnetic resonance imaging tracer, we used convection-enhanced delivery to perfuse rat and primate brainstems with IL13-PE and gadolinium-bound albumin (Gd-albumin). EXPERIMENTAL DESIGN: Thirty rats underwent convective brainstem perfusion of IL13-PE (0.25, 0.5, or 10 microg/mL) or vehicle. Twelve primates underwent convective brainstem perfusion of either IL13-PE (0.25, 0.5, or 10 microg/mL; n = 8), co-infusion of 125I-IL13-PE and Gd-albumin (n = 2), or co infusion of IL13-PE (0.5 microg/mL) and Gd-albumin (n = 2). The animals were permitted to survive for up to 28 days before sacrifice and histologic assessment. RESULTS: Rats showed no evidence of toxicity at all doses. Primates showed no toxicity at 0.25 or 0.5 microg/mL but showed clinical and histologic toxicity at 10 microg/mL. Quantitative autoradiography confirmed that Gd-albumin precisely tracked IL13-PE anatomic distribution and accurately showed the volume of distribution. CONCLUSIONS: IL13-PE can be delivered safely and effectively to the primate brainstem at therapeutic concentrations and over clinically relevant volumes using convection-enhanced delivery. Moreover, the distribution of IL13-PE can be accurately tracked by co-infusion of Gd-albumin using real-time magnetic resonance imaging. PMID- 16707616 TI - Immunogenic HLA-B*0702-restricted epitopes derived from human telomerase reverse transcriptase that elicit antitumor cytotoxic T-cell responses. AB - PURPOSE: The human telomerase reverse transcriptase (hTERT) is considered as a potential target for cancer immunotherapy because it is preferentially expressed in tumor cells. To increase the applicability of hTERT-based immunotherapy, we set out to identify CTL epitopes in hTERT restricted by HLA-B*0702 molecule, a common MHC class I allele. EXPERIMENTAL DESIGN: HLA-B*0702-restricted peptides from hTERT were selected by using a method of epitope prediction and tested for their immunogenicity in human (in vitro) and HLA-B*0702 transgenic mice (in vivo). RESULTS: All the six hTERT peptides that were predicted to bind to HLA B*0702 molecule were found to induce primary human CTL responses in vitro. The peptide-specific CD8+ CTL lines were tested against various hTERT+ tumor cells. Although differences were observed according to the tumor origin, only three CTL lines specific for p277, p342, and p351 peptides exhibited cytotoxicity against tumor cells in a HLA-B*0702-restricted manner. In addition, this cytotoxicity was inhibited by the addition of peptide-loaded cold target cells and indicated that these epitopes are naturally processed and presented on the tumor cells. Further, in vivo studies using humanized HLA-B*0702 transgenic mice showed that all the candidate peptides were able to induce CTL responses after peptide immunization. Furthermore, vaccination with a plasmid DNA encoding full-length hTERT elicited peptide-specific CTL responses, indicating that these epitopes are efficiently processed in vivo. CONCLUSIONS: Together with previously reported hTERT epitopes, the identification of new CTL epitopes presented by HLA-B*0702 increases the applicability of hTERT-based immunotherapy to treating cancer. PMID- 16707617 TI - Novel cytosine deaminase fusion gene enhances the effect of radiation on breast cancer in bone by reducing tumor burden, osteolysis, and skeletal fracture. AB - BACKGROUND: Painful breast carcinoma metastases in bone are a common manifestation of malignant disease. Eradication of these tumors can be evasive, and as a result, skeletal morbidity increases with disease progression. EXPERIMENTAL DESIGN: The treatment potential of cytosine deaminase (CD) gene therapy combined with radiation treatment was evaluated in vitro and in vivo using a 4T1 murine breast carcinoma model. 4T1 carcinoma cells were transduced with a fusion gene encoding the extracellular and transmembrane domains of the human nerve growth factor receptor and the cytoplasmic portion of the yeast CD gene (NGFR-CD(y)). RESULTS AND CONCLUSIONS: CD-expressing tumor cells (4TCD(y)) were highly sensitive to treatment by 5-fluorocytosine prodrug (P < 0.0001). 5 Fluorocytosine treatment of 4TCD(y), but not 4T1 cells, enhanced the effects of radiation in vitro (P < 0.0001). 5-Fluorocytosine prodrug treatment also increased the therapeutic potential of radiation in vivo. Mice with 4TCD(y) intrafemoral tumors showed increased effectiveness of radiation based on improved reductions in tumor size, reductions in tumorigenic osteolysis, and a decrease in skeletal fractures (P < 0.01). PMID- 16707618 TI - Evaluation of toxicity following electrically mediated interleukin-12 gene delivery in a B16 mouse melanoma model. AB - PURPOSE: Interleukin-12 (IL-12) has potential as an immunotherapeutic agent for the treatment of cancer but is unfortunately associated with toxicity. Delivery of a plasmid encoding IL-12 with electroporation induces an antitumor effect in the B16 mouse melanoma model without serious side effects. To translate this observation to the clinic, an evaluation of toxicity was done in the mouse model. EXPERIMENTAL DESIGN: Weight change, tumor response, blood chemistry and hematology values, and serum IL-12 levels were evaluated. Multiple tissues were analyzed histopathologically. RESULTS: A pronounced reduction in tumor volume, including a large percentage of complete regressions, was observed after electrically mediated gene therapy. No significant increases in serum IL-12 levels were detected. Tumor-bearing mice showed an increased number of atypical hematology values when compared with normal naive controls. Statistically significant differences in chemistry and hematology values were observed sporadically in most of the standard chemistry and hematology categories in all groups. The only histopathologic abnormality specific to the animals receiving both plasmid and electroporation was inflammation associated with the kidney at the last time point. CONCLUSIONS: In general, mice that received both plasmid and electroporation showed the least abnormal histopathologic findings and were found to be in the best health, reflecting the reduced burden of disease. No significant toxic effects due to the IL-12 gene therapy were observed. PMID- 16707619 TI - Telomerase template antagonist GRN163L disrupts telomere maintenance, tumor growth, and metastasis of breast cancer. AB - PURPOSE: Maintenance of telomeres by telomerase is critical for the continuing proliferation of most advanced cancer cells. Telomerase activity has been detected in the vast majority of cancer cells but not most normal cells, making the enzyme an attractive target for anticancer therapy. The aim of this study was to address the breast cancer translational potential of the novel telomerase inhibitor, GRN163L. EXPERIMENTAL DESIGN: In the present study, we investigated the effects of GRN163L treatment on a panel of breast cancer cells representing different tumor subtypes with varying genetic backgrounds, including ER+, ER-, HER2+, BRCA1 mutant breast tumor cells as well as doxorubicin-resistant cancer cells. To investigate the in vivo effects of GRN163L, we employed a breast cancer xenograft and metastasis model that simulates a clinical situation in which a patient arrives with a primary tumor that may be then treated or surgically removed. RESULTS: GRN163L effectively inhibited telomerase activity in a dose dependent fashion in all breast cancer cell lines resulting in progressive telomere shortening. A mismatch control oligonucleotide showed no effect on telomerase activity and GRN163L did not significantly affect telomere shortening in normal human mammary epithelial cells or in endothelial cells. Breast cancer cells that exhibited telomerase inhibition also exhibited significant reduction in colony formation and tumorigenicity. Furthermore, GRN163L suppressed tumor growth and lung metastases (P = 0.017) of MDA-MB-231 cells in vivo after 4 weeks of treatment. CONCLUSIONS: These results show in vivo effectiveness of GRN163L in breast cancer and support its promising clinical potential for breast cancer treatment. PMID- 16707620 TI - Liposomal quercetin efficiently suppresses growth of solid tumors in murine models. AB - PURPOSE: Quercetin is a potent chemotherapeutic drug. Clinical trials exploring different schedules of administration of quercetin have been hampered by its extreme water insolubility. To overcome this limitation, this study is aimed to develop liposomal quercetin and investigate its distribution in vivo and antitumor efficacy in vivo and in vitro. EXPERIMENTAL DESIGN: Quercetin was encapsulated in polyethylene glycol 4000 liposomes. Biodistribution of liposomal quercetin i.v. at 50 mg/kg in tumor-bearing mice was detected by high-performance liquid chromatography. Induction of apoptosis by liposomal quercetin in vitro was tested. The antitumor activity of liposomal quercetin was evaluated in the immunocompetent C57BL/6N mice bearing LL/2 Lewis lung cancer and in BALB/c mice bearing CT26 colon adenocarcinoma and H22 hepatoma. Tumor volume and survival time were observed. The mechanisms underlying the antitumor effect of quercetin in vivo was investigated by detecting the microvessel density, apoptosis, and heat shock protein 70 expression in tumor tissues. RESULTS: Liposomal quercetin could be dissolved in i.v. injection and effectively accumulate in tumor tissues. The half-time of liposomal quercetin was 2 hours in plasma. The liposomal quercetin induced apoptosis in vitro and significantly inhibited tumor growth in vivo in a dose-dependent manner. The optimal dose of liposomal quercetin resulted in a 40-day survival rate of 40%. Quantitative real-time PCR showed that liposomal quercetin down-regulated the expression of heat shock protein 70 in tumor tissues. Immunohistochemistry analysis showed that liposomal quercetin inhibited tumor angiogenesis as assessed by CD31 and induced tumor cell apoptosis. CONCLUSIONS: Our data indicated that pegylated liposomal quercetin can significantly improve the solubility and bioavailability of quercetin and can be a potential application in the treatment of tumor. PMID- 16707621 TI - Tumor-targeting properties of novel antibodies specific to the large isoform of tenascin-C. AB - BACKGROUND: The targeted delivery of bioactive molecules with antibodies specific to tumor-associated antigens represents a promising strategy for improving the efficacy of tumor therapy. The large isoform of tenascin-C, an abundant glycoprotein of the tumor extracellular matrix, is strongly overexpressed in adult tissue undergoing tissue remodeling, including wound healing and neoplasia, and has been implicated in a variety of different cancers while being virtually undetectable in most normal adult tissues. EXPERIMENTAL DESIGN: We have used antibody phage technology to generate good-quality human recombinant antibodies (F16 and P12) specific to the alternatively spliced domains A1 and D of the large isoform of tenascin-C. The tumor-targeting properties of F16 and P12 were assessed by biodistribution studies in tumor xenografts using the antibodies in small immunoprotein (SIP) format. RESULTS: SIP(F16) selectively accumulated at the tumor site with 4.5%ID/g at 24 hours in the U87 glioblastoma model but was rapidly cleared from other organs (tumor-to-organ ratios, approximately 10:1). The accumulation of SIP(P12) in the tumor was lower compared with SIP(F16) and persistent levels of radioactivity were observed in the intestine. CONCLUSIONS: These data suggest that the F16 antibody, specific to domain A1 of tenascin-C, is a promising building block for the development of antibody-based pharmaceuticals in view of its excellent tumor-targeting performance and the strong expression of the antigen in a variety of primary and metastatic tumors. PMID- 16707622 TI - Frequency and spectrum of cancers in the Peutz-Jeghers syndrome. AB - BACKGROUND: Although an increased cancer risk in Peutz-Jeghers syndrome is established, data on the spectrum of tumors associated with the disease and the influence of germ-line STK11/LKB1 (serine/threonine kinase) mutation status are limited. EXPERIMENTAL DESIGN: We analyzed the incidence of cancer in 419 individuals with Peutz-Jeghers syndrome, and 297 had documented STK11/LKB1 mutations. RESULTS: Ninety-six cancers were found among individuals with Peutz Jeghers syndrome. The risk for developing cancer at ages 20, 30, 40, 50, 60, and 70 years was 2%, 5%, 17%, 31%, 60%, and 85%, respectively. The most common cancers represented in this analysis were gastrointestinal in origin, gastroesophageal, small bowel, colorectal, and pancreatic, and the risk for these cancers at ages 30, 40, 50, and 60 years was 1%, 9%, 15%, and 33%, respectively. In women with Peutz-Jeghers syndrome, the risk of breast cancer was substantially increased, being 8% and 31% at ages 40 and 60 years, respectively. Kaplan-Meier analysis showed that cancer risks were similar in Peutz-Jeghers syndrome patients with identified STK11/LKB1 mutations and those with no detectable mutation (log rank test of difference chi2 = 0.62; 1 df; P = 0.43). Furthermore, the type or site of STK11/LKB1 mutation did not significantly influence cancer risk. CONCLUSIONS: The results from our study provide quantitative information on the spectrum of cancers and risks of specific cancer types associated with Peutz Jeghers syndrome. PMID- 16707623 TI - Effects of Helicobacter pylori eradication on methylation status of E-cadherin gene in noncancerous stomach. AB - PURPOSE: Promoter hypermethylation of E-cadherin plays an important role on gastric cancer development. Whereas E-cadherin methylation was frequently detected in the stomach of Helicobacter pylori-infected individuals, we tested whether eradication of H. pylori alters the methylation status of the noncancerous gastric epithelium. EXPERIMENTAL DESIGN: Endoscopic biopsies were taken from the antrum and corpus of H. pylori-infected subjects without gastric cancer. Presence of methylated E-cadherin sequences in the gastric specimens was detected by methylation-specific PCR. Bisulfite DNA sequencing was done to determine the topographical distribution and changes in methylation profiles with H. pylori eradication. RESULTS: Among the 28 H. pylori-infected subjects (median age, 44.5 years), 15 (53.6%) had E-cadherin methylation detected in stomach at baseline. Discordant methylation patterns between the antrum and corpus were noted in six patients. One year after successful H. pylori eradication, there was a significant reduction in the methylation density of the promoter region and exon 1 of the E-cadherin gene as detected by bisulfite DNA sequencing (P < 0.001). CONCLUSION: Promoter methylation in E-cadherin was frequently detected in the stomach of H. pylori-infected individuals. Eradication of H. pylori might possibly reduce the methylation density in E-cadherin gene and the chance of subsequent neoplastic transformation. PMID- 16707624 TI - Heterogeneous nuclear ribonucleoprotein G shows tumor suppressive effect against oral squamous cell carcinoma cells. AB - PURPOSE: Heterogeneous nuclear ribonucleoproteins (hnRNP) are nucleic acid binding proteins involved in RNA processing. We found that hnRNP G is expressed in normal human oral epithelial cells while frequently not found in the cells derived from human oral squamous cell carcinomas (HOSCC). The current study was designed to test the hypothesis that hnRNP G is a tumor suppressor. EXPERIMENTAL DESIGN: We investigated the expression levels of hnRNP G protein in normal, precancerous, and malignant oral tissues by in situ immunohistochemistry. In addition, wild-type or mutant hnRNP G was ectopically overexpressed in HOSCC cells and their effects on cellular replication kinetics, colonogenic efficiency, anchorage-independent growth, and in vivo tumorigenicity were determined. RESULTS: In situ immunohistochemical staining showed robust presence of hnRNP G in the basal cell layers of normal oral epithelium but the level of its staining was markedly reduced in dysplastic or cancerous tissues. Ectopic expression of wild-type hnRNP G in cancer cells lacking hnRNP G expression or containing mutant hnRNP G resulted in severe retardation of proliferation, reduction of colonogenic efficiency, loss of anchorage-independent growth, and reduction of in vivo tumorigenicity in immunocompromised mice. In addition, hnRNP G overexpression led to up-regulation of the expression of TXNIP, a cell cycle inhibitory gene, and significantly reduced the expression of the genes that promote cellular proliferation, such as EGR1, JUND, JUNB, FOS, FOSL1, ROS, and KIT. CONCLUSIONS: These results indicate that hnRNP G is a tumor suppressor against HOSCC but its mechanisms of action remain to be further investigated. PMID- 16707625 TI - Transforming growth factor-beta receptor type 1 (TGFbetaRI) kinase activity but not p38 activation is required for TGFbetaRI-induced myofibroblast differentiation and profibrotic gene expression. AB - Transforming growth factor-beta (TGFbeta) is a major mediator of normal wound healing and of pathological conditions involving fibrosis, such as idiopathic pulmonary fibrosis. TGFbeta also stimulates the differentiation of myofibroblasts, a hallmark of fibrotic diseases. In this study, we examined the underlying processes of TGFbetaRI kinase activity in myofibroblast conversion of human lung fibroblasts using specific inhibitors of TGFbetaRI (SD-208) and p38 mitogen-activated kinase (SD-282). We demonstrated that SD-208, but not SD-282, inhibited TGFbeta-induced SMAD signaling, myofibroblast transformation, and collagen gel contraction. Furthermore, we extended our findings to a rat bleomycin-induced lung fibrosis model, demonstrating a significant decrease in the number of myofibroblasts at fibroblastic foci in animals treated with SD-208 but not those treated with SD-282. SD-208 also reduced collagen deposition in this in vivo model. Microarray analysis of human lung fibroblasts identified molecular fingerprints of these processes and showed that SD-208 had global effects on reversing TGFbeta-induced genes involved in fibrosis, inflammation, cell proliferation, cytoskeletal organization, and apoptosis. These studies also revealed that although the p38 pathway may not be needed for appearance or disappearance of the myofibroblast, it can mediate a subset of inflammatory and fibrogenic events of the myofibroblast during the process of tissue repair and fibrosis. Our findings suggest that inhibitors such as SD-208 may be therapeutically useful in human interstitial lung diseases and pulmonary fibrosis. PMID- 16707626 TI - A tail of two signals: the C terminus of the A(2A)-adenosine receptor recruits alternative signaling pathways. AB - G protein-coupled receptors are endowed with carboxyl termini that vary greatly in length and sequence. In most instances, the distal portion of the C terminus is dispensable for G protein coupling. This is also true for the A(2A)-adenosine receptor, where the last 100 amino acids are of very modest relevance to G(s) coupling. The C terminus was originally viewed mainly as the docking site for regulatory proteins of the beta-arrestin family. These beta-arrestins bind to residues that have been phosphorylated by specialized kinases (G protein-coupled receptor kinases) and thereby initiate receptor desensitization and endocytosis. More recently, it has become clear that many additional "accessory" proteins bind to C termini of G protein-coupled receptors. The article by Sun et al. in the current issue of Molecular Pharmacology identifies translin-associated protein-X as yet another interaction partner of the A(2A) receptor; translin-associated protein allows the A(2A) receptor to impinge on the signaling mechanisms by which p53 regulates neuronal differentiation, but the underlying signaling pathways are uncharted territory. With a list of five known interaction partners, the C terminus of the A(2A) receptor becomes a crowded place. Hence, there must be rules that regulate the interaction. This allows the C terminus to act as coincidence detector and as signal integrator. Despite our ignorance about the precise mechanisms, the article has exciting implications: the gene encoding for translin-associated protein-X maps to a locus implicated in some forms of schizophrenia; A(2A) receptor agonists are candidate drugs for the treatment of schizophrenic symptoms. It is of obvious interest to explore a possible link. PMID- 16707627 TI - Cross-talk between G(s)- and G(q)-coupled pathways in regulation of interleukin-4 by A(2B) adenosine receptors in human mast cells. AB - Human mast cells express functional A(2A) and A(2B) adenosine receptors. However, only stimulation of A(2B), not A(2A), leads to secretion of interleukin (IL)-4, an important step in adenosine receptor-mediated induction of IgE synthesis by B cells. In this study, we investigate intracellular pathways that link stimulation of A(2B) receptors to IL-4 up-regulation in HMC-1 mast cells. Both A(2A) and A(2B) receptors couple to G(s) proteins and stimulate adenylate cyclase, but only A(2B) stimulates phospholipase Cbeta through coupling to G(q) proteins leading to activation of protein kinase C and calcium mobilization. Inhibition of phospholipase Cbeta completely blocked A(2B) receptor-dependent IL-4 secretion. The protein kinase C inhibitor 2-{8-[(dimethylamino)-methyl]-6,7,8,9 tetrahydropyrido[1,2-a]indol-3-yl}-3-(1-methyl-1H-indol-3-yl)maleimide (Ro-32 0432) had no effect on A(2B) receptor-mediated IL-4 secretion but inhibited phorbol 12-myristate 13-acetate-stimulated IL-4 secretion. In contrast, chelation of intracellular Ca(2+) inhibited both A(2B) receptor- and ionomycin-dependent IL 4 secretion. This Ca(2+)-sensitive pathway probably includes calcineurin and nuclear factor of activated T cells, because A(2B) receptor-dependent IL-4 secretion was blocked with cyclosporin A or 11R-VIVIT peptide. G(s)-linked pathways also play a role in the A(2B) receptor-dependent stimulation of IL-4 secretion; inhibition of adenylate cyclase or protein kinase A attenuated A(2B) receptor-dependent IL-4 secretion. Although stimulation of adenylate cyclase with forskolin did not increase IL-4 secretion on its own, it potentiated the effect of Pasteurella multocida toxin by 2-fold and ionomycin by 3-fold. Both forskolin and stimulation of A(2B) receptors up-regulated NFATc1 protein levels. We conclude that A(2B) receptors up-regulate IL-4 through G(q) signaling that is potentiated via cross-talk with G(s)-coupled pathways. PMID- 16707628 TI - Neutrophil-mediated eosinophil transmigration: parting the Red Sea or leaving bread crumbs. PMID- 16707629 TI - Biomechanical risk factors associated with neuropathic ulceration of the hallux in people with diabetes mellitus. AB - In this study of people with diabetes mellitus and peripheral neuropathy, it was found that the feet of patients with a history of hallux ulceration were more pronated and less able to complete a single-leg heel rise compared with the feet of patients with a history of ulceration elsewhere on the foot. The range of active first metatarsophalangeal joint dorsiflexion was found to be significantly lower in the affected foot. Ankle dorsiflexion, subtalar joint range of motion, and angle of gait differed from normal values but were similar to those found in other studies involving diabetic subjects and were not important factors in the occurrence of hallux ulceration. These data indicate that a more pronated foot type is associated with hallux ulceration in diabetic feet. Further studies are required to evaluate the efficacy of footwear and orthoses in altering foot posture to manage hallux ulceration. PMID- 16707630 TI - Trephine arthrodesis of the foot and ankle: indications, operative technique, and long-term follow-up. AB - We conducted a retrospective analysis of trephine arthrodesis in a total of 22 joints in 15 feet to determine the efficacy of this procedure in the foot and ankle. Twelve of the 22 joints that were fused involved the tarsometatarsal articulations, 3 the naviculocuneiform joint, 6 the subtalar joint, and 1 the ankle. A successful fusion rate of 95% (21/22) was achieved. These results lend evidence that the trephine arthrodesis technique can be used with satisfactory results and confidence in the foot and ankle. PMID- 16707631 TI - Effective orthotic therapy for the painful cavus foot: a randomized controlled trial. AB - Patients with a cavus or high-arched foot frequently experience foot pain, which can lead to significant limitation in function. Custom foot orthoses are widely prescribed to treat cavus foot pain. However, no clear guidelines for their construction exist, and there is limited evidence of their efficacy. In a randomized, single-blind, sham-controlled trial, the effect of custom foot orthoses on foot pain, function, quality of life, and plantar pressure loading in people with a cavus foot type was investigated. One hundred fifty-four participants with chronic musculoskeletal foot pain and bilateral cavus feet were randomly assigned to a treatment group receiving custom foot orthoses (n = 75) or to a control group receiving simple sham insoles (n = 79). At 3 months, 99% of the participants provided follow-up data using the Foot Health Status Questionnaire. Foot pain scores improved more with custom foot orthoses than with the control (difference, 8.3 points; 95% confidence interval [CI], 1.2 to 15.3 points; P = .022). Function scores also improved more with custom foot orthoses than with the control (difference, 9.5 points; 95% CI, 2.9 to 16.1 points; P = .005). Quality-of-life data favored custom foot orthoses, although differences reached statistical significance only for physical functioning (difference, 7.0 points; 95% CI, 1.9 to 12.1 points; P = .008). Plantar pressure improved considerably more with custom foot orthoses than with the control for all regions of the foot (difference, -3.0 N . s/cm(2); 95% CI, -3.7 to -2.4 N . s/cm(2); P < .001). In conclusion, custom foot orthoses are more effective than a control for the treatment of cavus foot pain and its associated limitation in function. PMID- 16707632 TI - The subtalar joint axis locator: a preliminary report. AB - A new clinical device, the subtalar joint axis locator, was created to track the three-dimensional location of the subtalar joint axis during weightbearing movements of the foot. The assumption was that if the anterior exit point of the subtalar joint axis is stationary relative to the dorsal aspect of the talar neck, then, by performing radiographs of the feet with the subtalar joint axis locator in place on the foot, the ability of the locator to track rotations and translations of the talar neck and thus the subtalar joint axis in space could be approximated. In this preliminary study of two adults, the subtalar joint axis locator accurately tracked the talar neck position during weightbearing rotational motions of the subtalar joint. The device was also used in a series of subjects to determine its dynamic capabilities. It is possible, then, that the subtalar joint axis locator can reliably track the spatial location of the subtalar joint axis during weightbearing movements of the foot. PMID- 16707633 TI - Intralesional bleomycin sulfate injection for the treatment of verruca plantaris. AB - Sixty-two patients were treated for single or multiple warts by intralesional injection of bleomycin sulfate (1.5 U/mL) and then were observed for 6 months. The dose varied according to the size of the lesion and ranged from 0.25 to 1.0 mL per injection per lesion, up to a maximum dose of 3 mL. The total cure rate was 87% after one or two injections. Twelve of the 62 patients required a second injection. PMID- 16707634 TI - Foot pathology in patients with Paget's disease of bone. AB - The characteristic bone deformities associated with advanced Paget's disease of bone may result in abnormal gait and plantar foot pressures, which contribute to the development of pressure-related skin problems. This study aimed to characterize the foot problems and foot-care needs in this patient group and to investigate the effect of disease distribution on the occurrence of foot pathology. One hundred thirty-four patients with Paget's disease were assessed clinically, and the Foot Structure Index was completed for each patient. Patients completed self-administered questionnaires concerning foot function (Foot Function Index) and quality of life (12-Item Short Form). The results of this study suggest that foot-health and footwear problems are common in patients with Paget's disease. Forty percent of the patients required professional foot care in addition to those already receiving it. The site of pagetic involvement did not affect the occurrence of foot pathology. Further research is required on the impact of the extent of any femoral or tibial deformity on foot symptoms and pathology. Biomechanical studies of the forces and motion in the foot related to different degrees of femoral and tibial deformity might also help determine the impact of deformity on foot pathology. PMID- 16707635 TI - A prospective analysis of the treatment of friction blisters with 2 octylcyanoacrylate. AB - Early military investigations of blister treatment using tissue adhesives have shown promise, finding that these agents provide pain relief, prevent infection, and allow continuation of recruit training. A trial was conducted to compare a 2 octylcyanoacrylate treatment with military standard therapy. Patients were recruited during foot evaluation at the end of a 10-km military road march. Seventy-five patients were screened, and 51 were enrolled. Twenty-six patients received 2-octylcyanoacrylate and 25 received standard treatment. A total of 80 blisters were treated (40 in each group). The mean (SD) blister size in the 2 octylcyanoacrylate group was 12.7 (6.2) mm and in the standard group was 12.0 (5.7) mm. There were no statistically significant differences in any of the baseline variables. Baseline and repeated-measures visual analog scale scores demonstrated no statistically significant differences in initial or repeat pain scores. Both groups showed a similar change across time, with a nonsignificant trend toward improvement in the standard therapy group at 10 min (28.5 versus 24.9) and in the 2-octylcyanoacrylate group at 3 days (42.9 versus 50.1). Mean Likert scores were similar, indicating no difference in patient satisfaction. Time to resumption of normal activity was similar, with one patient in each group unable to return to activity at the time of follow-up. There was a trend toward an increased proportion of patients in the 2-octylcyanoacrylate group who were able to return to normal activity within 48 hours, but this did not reach statistical significance. 2-Octylcyanoacrylate was associated with a greater degree of procedural discomfort. No infected blisters were noted in either group. PMID- 16707636 TI - Relative metatarsal protrusion in the adult: a preliminary study. AB - We performed a bibliographic review of the systems proposed by various researchers to evaluate physiologic metatarsal protrusion. The system of measurement devised by Hardy and Clapham to evaluate the protrusion between the first and second metatarsals was adapted to study the whole metatarsal parabola. We studied the five metatarsals of 52 normal feet. Mean metatarsal protrusion relative to the second metatarsal was +1.21% for the first metatarsal, -3.84% for the third metatarsal, -9.66% for the fourth metatarsal, and -16.91% for the fifth metatarsal. PMID- 16707637 TI - Current concepts in diabetic microvascular dysfunction. AB - Microvascular dysfunction is an important component of the pathologic processes that occur in diabetic foot disease. The endothelial abnormalities observed in patients with diabetes mellitus are poorly understood, and evidence suggests that endothelial dysfunction could be involved in the pathogenesis of diabetic macroangiopathy and microangiopathy. With the advent of insulin replacement in the early 1900s and increased efforts toward metabolic control of diabetes, long term complications of this disease have become apparent. These late-term complications are primarily disorders of the vascular system. This article reviews the process of microvascular dysfunction and how it may relate to the pathogenesis of diabetic foot problems. PMID- 16707638 TI - Acquired constriction ring syndrome. AB - Acquired constriction ring syndrome is a clinical condition of infancy characterized by circumferential constriction of a toe or another appendage, such as fingers and genitalia. The foot and ankle specialist should be aware of this condition because vascular obstruction of the affected appendage can rapidly lead to gangrene and autoamputation. Treatment consists of prompt identification and removal of the constricting foreign material. Although this condition is uncommon, it can lead to digital loss. Early treatment yields a good prognosis. A case report is presented of a 9-week-old infant who experienced acquired constriction ring syndrome caused by a strand of hair wrapped around the third and fourth toes that was treated by unwinding the hair under loupe magnification. PMID- 16707639 TI - Talar dome fracture repaired using bioabsorbable fixation. AB - We describe a man with an acute osteochondral defect in the lateral talar dome associated with a supination-adduction-type ankle fracture. The osteochondral defect was readily visible on plain film radiographs, and magnetic resonance imaging was ordered to determine the full extent of soft-tissue and articular injury. It was discovered that the patient had a stage IV lesion of the talar dome, with complete inversion of the fragment, and rupture of the anterior talofibular and calcaneofibular lateral ankle ligaments. Furthermore, the patient experienced an oblique fracture of the medial malleolus with comminution. The talar dome lesion was surgically reduced and fixated using bioabsorbable pins. Nine months after surgery, the patient was fully recovered from his injury and had no functional limitations. PMID- 16707640 TI - Arthroscopic management of talar dome lesions using a transmalleolar approach. AB - Surgical treatment of posteromedial talar dome lesions is frequently necessary for Berndt and Harty grade IV osteochondral defects and nondisplaced osteochondral fragments resistant to conservative modalities. When operative intervention is indicated, the approach and management can be complicated by the location and extent of the injury. The operative technique we advocate allows direct exposure of the lesion and minimizes damage to healthy articular cartilage and surrounding soft tissue. Use of a drill guide assists the surgeon in precisely placing a transmalleolar portal through the tibia for subchondral drilling of osteochondral defects when the lesions are inaccessible through traditional arthroscopic portals. PMID- 16707641 TI - Wound-care resources on the Internet: a second update. AB - An updated selection of high-quality Internet resources related to wound and ulcer care is presented. Of potential use to the podiatric medical practitioner, educator, resident, and student, some Web sites that cover hyperbaric medicine, antibiotic use, and wound and ulcer prevention are also included. These Web sites have been evaluated on the basis of their potential to enhance the practice of podiatric medicine, in addition to contributing to the educational process. Readers who require a quick reference source to wound and ulcer care may find this report useful. PMID- 16707642 TI - Effectiveness of extracorporeal shockwave treatment in 353 patients with chronic plantar fasciitis. PMID- 16707643 TI - Effectiveness of extracorporeal shockwave treatment in 353 patients with chronic plantar fasciitis. PMID- 16707648 TI - Distribution of influenza vaccine to high-risk groups. AB - Vaccine distribution programs have historically targeted individuals at high risk of complications due to influenza. Despite recommendations from the Advisory Committee on Immunization Practices, vaccination coverage among high-risk populations has been generally low. This review systematically summarizes the recent literature evaluating programs in different settings, from within medical settings to venue-based and community-based approaches, in an effort to identify successful program components. The published literature was identified by using the MEDLINE database from 1990 to 2006 covering studies that reported on interventions or programs aimed at vaccinating high-risk populations. The authors reviewed 56 studies. In the United States, the Healthy People 2010 goals included 90% vaccination coverage for adults aged > or = 65 years and 60% for high-risk adults aged 18-64 years. Only a handful of the studies reviewed managed to meet those goals. Interventions that increased vaccination coverage to Healthy People 2010 goals included advertising, provider and patient mailings, registry-based telephone calls, patient and staff education, standing orders coupled with standardized forms, targeting of syringe exchange customers, and visiting nurses. Few studies evaluated the impact of vaccination programs by race/ethnicity and socioeconomic status. Few studies targeted individuals outside of the health-care and social services sectors. Given the growing disparities in health and health care access, understanding the way in which interventions can remedy disparities is crucial. PMID- 16707649 TI - XRCC3 and XPD/ERCC2 single nucleotide polymorphisms and the risk of cancer: a HuGE review. AB - Hundreds of polymorphisms in DNA repair genes have been identified; however, for many of these polymorphisms, the impact on repair phenotype and cancer susceptibility remains uncertain. In this review, the authors focused on the x ray repair cross-complementing protein group 3 (XRCC3) and xeroderma pigmentosum group D (XPD)/excision repair cross-complementing rodent repair deficiency (ERCC2) genes, because they are among the most extensively studied but no final conclusion has yet been drawn about their role in cancer occurrence. XRCC3 participates in DNA double-strand break/recombinational repair through homologous recombination to maintain chromosome stability. XPD/ERCC2 is a helicase involved in the nucleotide excision repair pathway, which recognizes and repairs many structurally unrelated lesions, such as bulky adducts and thymidine dimers. The authors identified a sufficient number of epidemiologic studies on cancer to perform meta-analyses for XPD/ERCC2 variants in codons 156, 312, and 751 and XRCC3 variants in codon 241. The authors evaluated all cancer sites to investigate whether DNA repair is likely to take place in a rather nonspecific manner for different carcinogens and different cancers. For the most part, the authors found no association between these genes and the cancer sites investigated, except for some statistically significant associations between XPD/ERCC2 single nucleotide polymorphisms and skin, breast, and lung cancers. PMID- 16707650 TI - A prospective cohort study of shift work and risk of ischemic heart disease in Japanese male workers. AB - This study prospectively examined the association between shift work and the risk of ischemic heart disease among Japanese male workers. A baseline survey, which involved 110,792 inhabitants (age range: 40-79 years) from 45 areas throughout Japan, was conducted between 1988 and 1990. The causes of death were identified from death certificates. The analysis was restricted to 17,649 men (age range: 40 59 years) who were employed at the time of the baseline survey. All subjects were asked to indicate the most regular shift work that they had undertaken previously: day work, rotating-shift work, or fixed-night work. The Cox proportional hazards model was used to estimate the risks of shift work for ischemic heart disease. During the 233,869 person-years of follow-up, a total of 1,363 deaths were recorded, 86 of which were due to ischemic heart disease. Compared with the day workers, the rotating-shift workers had a significantly higher risk of death due to ischemic heart disease (relative risk = 2.32, 95% confidence interval: 1.37, 3.95; p = 0.002), whereas fixed-night work was not associated with ischemic heart disease (relative risk = 1.23, 95% confidence interval: 0.49, 3.10; p = 0.658). In addition, subjects with coronary risk factors, such as hypertension, overweight, habitual alcohol consumption, and smoking, were highly susceptible to the effect of rotating-shift work on the risk of death due to ischemic heart disease. PMID- 16707651 TI - The relation between components of adult height and intimal-medial thickness in middle age: the Atherosclerosis Risk in Communities Study. AB - The authors aimed to investigate the relation between components of adult height (leg and trunk length) and atherosclerosis in middle age, using data from 12,254 participants (aged 44-65 years) in the Atherosclerosis Risk in Communities (ARIC) Study. Intimal-medial thickness (IMT) as measured by B-mode ultrasound was the outcome, and exposures were trunk and leg lengths as estimated (using sitting height and the difference between sitting and standing height) at the first study examination in 1987-1989. The mean IMT was 0.73 (standard deviation, 0.17) mm. Greater leg length was associated with lower IMT, with the largest difference being for Black men (a 0.045 (95% confidence interval: 0.023, 0.068)-mm lower IMT per 10-cm higher leg length). Greater trunk length was associated with higher IMT, with the largest difference being for White men (a 0.024 (95% confidence interval: 0.005, 0.044)-mm higher IMT per 10-cm higher trunk length). Although the effect sizes were small, leg length was inversely associated with atherosclerosis, consistent with the results of other studies with cardiovascular disease outcomes. PMID- 16707652 TI - Invited commentary: on the road to improved exposure assessment using geographic information systems. PMID- 16707653 TI - Measuring screening intensity in case-control studies of the efficacy of mammography. AB - Of great interest in studies of screening for breast cancer is the relative efficacy of different screening frequencies (intensities). Prior work has suggested that estimates of the association between screening intensity and outcome in case-control studies would not produce valid results and that only binary indicators (no screens vs. one or more) of exposure can be used. Using case-control studies drawn from simulated cohorts of 30,000-40,000 women, the authors found that biases demonstrated in prior studies can be explained by 1) misclassification of true exposure groups by observed screening history, and 2) differential exposure misclassification of cases and controls. Binary as well as ordered categorical and interval measures can be biased unless they account for misclassification. By combining measurements of screening history from multiple periods of observation of varying lengths and using repeated-measures logistic regression models, the effect of screening intensity can be estimated in the presence of misclassification. Assessing the effect of screening intensity in case-control studies of mammography is possible if principles and methods for misclassification and measurement error guide the analysis. PMID- 16707654 TI - Invited commentary: Considerations about specificity of associations, causal pathways, and heterogeneity in multilevel thinking. PMID- 16707655 TI - Urban neighborhood context, educational attainment, and cognitive function among older adults. AB - Existing research has not addressed the potential impact of neighborhood context- educational attainment of neighbors in particular--on individual-level cognition among older adults. Using hierarchical linear modeling, the authors analyzed data from the 1993 Study of Assets and Health Dynamics Among the Oldest Old (AHEAD), a large, nationally representative sample of US adults born before 1924. Data from participants residing in urban neighborhoods (n = 3,442) were linked with 1990 US Census tract data. Findings indicate that 1) average cognitive function varies significantly across US Census tracts; 2) older adults living in low-education areas fare less well cognitively than those living in high-education areas, net of individual characteristics, including their own education; 3) this association is sustained when controlling for contextual-level median household income; and 4) the effect of individual-level educational attainment differs across neighborhoods of varying educational profiles. Promoting educational attainment among the general population living in disadvantaged neighborhoods may prove cognitively beneficial to its aging residents because it may lead to meliorations in stressful life conditions and coping deficiencies. PMID- 16707656 TI - Prognostic value of a novel classification scheme for heart failure: the Minnesota Heart Failure Criteria. AB - The authors present the Minnesota Heart Failure Criteria (MHFC), derived using latent class analysis from widely available items in the Framingham Criteria. The authors used 1995 and 2000 data on hospitalized Minnesota Heart Survey subjects discharged after myocardial infarction or heart failure (N = 7,379). Selected Framingham Criteria variables (dyspnea, pulmonary rales, cardiomegaly, interstitial or pulmonary edema on chest radiograph, S(3) heart sound, tachycardia) plus left ventricular ejection fraction were used. The discriminatory power of the MHFC was evaluated using age- and sex-adjusted 2-year mortality. A five-class latent class analysis model was collapsed into cases and noncases. Mortality estimates discriminated noncases (18%) from cases (43%) (p < 0.001). The MHFC performed better than previous truncated criteria (Framingham Criteria: 26% noncases, 43% cases; Duke Criteria: 29%, 40%; Killip Score: 31%, 44%; Boston Score: 28%, 45%). In a subset of patients admitted for heart failure (n = 5,128), the MHFC identified all but 2% (116/4,746) of cases found with a nearly full version of the Framingham Criteria. In terms of prognostic value, the MHFC are as precise as or more precise than several previous sets of truncated criteria. They closely approximate a nearly full version of the Framingham Criteria but require many fewer variables and can facilitate epidemiologic case finding for heart failure. PMID- 16707658 TI - Tests of rRNA hybridization to microarrays suggest that hybridization characteristics of oligonucleotide probes for species discrimination cannot be predicted. AB - Hybridization of rRNAs to microarrays is a promising approach for prokaryotic and eukaryotic species identification. Typically, the amount of bound target is measured by fluorescent intensity and it is assumed that the signal intensity is directly related to the target concentration. Using thirteen different eukaryotic LSU rRNA target sequences and 7693 short perfect match oligonucleotide probes, we have assessed current approaches for predicting signal intensities by comparing Gibbs free energy (DeltaG degrees) calculations to experimental results. Our evaluation revealed a poor statistical relationship between predicted and actual intensities. Although signal intensities for a given target varied up to 70-fold, none of the predictors were able to fully explain this variation. Also, no combination of different free energy terms, as assessed by principal component and neural network analyses, provided a reliable predictor of hybridization efficiency. We also examined the effects of single-base pair mismatch (MM) (all possible types and positions) on signal intensities of duplexes. We found that the MM effects differ from those that were predicted from solution-based hybridizations. These results recommend against the application of probe design software tools that use thermodynamic parameters to assess probe quality for species identification. Our results imply that the thermodynamic properties of oligonucleotide hybridization are by far not yet understood. PMID- 16707657 TI - Vapor, dust, and smoke exposure in relation to adult-onset asthma and chronic respiratory symptoms: the Singapore Chinese Health Study. AB - Occupational factors contribute to a significant fraction of respiratory disease and symptoms. The authors evaluated the role of occupational exposures in asthma, chronic bronchitis, and respiratory symptoms in the Singapore Chinese Health Study, a population-based cohort of adults aged 45-74 years at enrollment in 1993 1998. Information on occupations and occupational exposures was collected at enrollment for 52,325 subjects for whom respiratory outcomes were obtained via follow-up interviews in 1999-2004. Exposure to dusts from cotton, wood, metal, minerals, and/or asbestos was associated with nonchronic cough and/or phlegm (odds ratio (OR) = 1.19, 95% confidence interval (CI): 1.08, 1.30), chronic bronchitis (OR = 1.26, 95% CI: 1.01, 1.57), and adult-onset asthma (OR = 1.14, 95% CI: 1.00, 1.30). Cotton dust was the major contributor to respiratory symptoms. Vapor exposure from chemical solvents, dyes, cooling oils, paints, wood preservatives, and/or pesticides was associated with nonchronic cough or phlegm (OR = 1.14, 95% CI: 1.03, 1.27), chronic dry cough (OR = 1.55, 95% CI: 1.19, 2.01), and adult-onset asthma (OR = 1.34, 95% CI: 1.15, 1.56). Chemical solvents, cooling oils, and pesticides were the major contributors to respiratory symptoms. These data support the role of occupational exposures in the etiology of respiratory illness in a population-based cohort in Singapore with a low prevalence of atopic illness. PMID- 16707659 TI - Improvements of rolling circle amplification (RCA) efficiency and accuracy using Thermus thermophilus SSB mutant protein. AB - Rolling circle amplification (RCA) of plasmid or genomic DNA using random hexamers and bacteriophage phi29 DNA polymerase has become increasingly popular in the amplification of template DNA in DNA sequencing. We have found that the mutant protein of single-stranded DNA binding protein (SSB) from Thermus thermophilus (Tth) HB8 enhances the efficiency of amplification of DNA templates. In addition, the TthSSB mutant protein increased the specificity of phi29 DNA polymerase. We have overexpressed the native and mutant forms of TthSSB protein in Escherichia coli and purified them to homogeneity. In vitro, these proteins were found to bind specifically to single-stranded DNA. Addition of TthSSB mutant protein to RCA halved the elongation time required for phi29 DNA polymerase to synthesize DNA fragments in RCA. Furthermore, the presence of the TthSSB mutant protein essentially eliminates nonspecific DNA products in RCA reactions. PMID- 16707660 TI - RNA aptamers selected against DNA polymerase beta inhibit the polymerase activities of DNA polymerases beta and kappa. AB - DNA polymerase beta (polbeta), a member of the X family of DNA polymerases, is the major polymerase in the base excision repair pathway. Using in vitro selection, we obtained RNA aptamers for polbeta from a variable pool of 8 x 10(12) individual RNA sequences containing 30 random nucleotides. A total of 60 individual clones selected after seven rounds were screened for the ability to inhibit polbeta activity. All of the inhibitory aptamers analyzed have a predicted tri-lobed structure. Gel mobility shift assays demonstrate that the aptamers can displace the DNA substrate from the polbeta active site. Inhibition by the aptamers is not polymerase specific; inhibitors of polbeta also inhibited DNA polymerase kappa, a Y-family DNA polymerase. However, the RNA aptamers did not inhibit the Klenow fragment of DNA polymerase I and only had a minor effect on RB69 DNA polymerase activity. Polbeta and kappa, despite sharing little sequence similarity and belonging to different DNA polymerase families, have similarly open active sites and relatively few interactions with their DNA substrates. This may allow the aptamers to bind and inhibit polymerase activity. RNA aptamers with inhibitory properties may be useful in modulating DNA polymerase activity in cells. PMID- 16707661 TI - Multiple start codons and phosphorylation result in discrete Rad52 protein species. AB - The sequence of the Saccharomyces cerevisiae RAD52 gene contains five potential translation start sites and protein-blot analysis typically detects multiple Rad52 species with different electrophoretic mobilities. Here we define the gene products encoded by RAD52. We show that the multiple Rad52 protein species are due to promiscuous choice of start codons as well as post-translational modification. Specifically, Rad52 is phosphorylated both in a cell cycle independent and in a cell cycle-dependent manner. Furthermore, phosphorylation is dependent on the presence of the Rad52 C terminus, but not dependent on its interaction with Rad51. We also show that the Rad52 protein can be translated from the last three start sites and expression from any one of them is sufficient for spontaneous recombination and the repair of gamma-ray-induced double-strand breaks. PMID- 16707662 TI - Refining multiple sequence alignments with conserved core regions. AB - Accurate multiple sequence alignments of proteins are very important to several areas of computational biology and provide an understanding of phylogenetic history of domain families, their identification and classification. This article presents a new algorithm, REFINER, that refines a multiple sequence alignment by iterative realignment of its individual sequences with the predetermined conserved core (block) model of a protein family. Realignment of each sequence can correct misalignments between a given sequence and the rest of the profile and at the same time preserves the family's overall block model. Large-scale benchmarking studies showed a noticeable improvement of alignment after refinement. This can be inferred from the increased alignment score and enhanced sensitivity for database searching using the sequence profiles derived from refined alignments compared with the original alignments. A standalone version of the program is available by ftp distribution (ftp://ftp.ncbi.nih.gov/pub/REFINER) and will be incorporated into the next release of the Cn3D structure/alignment viewer. PMID- 16707663 TI - Thermodynamic characterization of an engineered tetracycline-binding riboswitch. AB - Riboswitches reflect a novel concept in gene regulation that is particularly suited for technological adaptation. Therefore, we characterized thermodynamically the ligand binding properties of a synthetic, tetracycline (tc) binding RNA aptamer, which regulates gene expression in a dose-dependent manner when inserted into the untranslated region of an mRNA. In vitro, one molecule of tc is bound by one molecule of partially pre-structured and conformationally homogeneous apo-RNA. The dissociation constant of 770 pM, as determined by fluorimetry, is the lowest reported so far for a small molecule-binding RNA aptamer. Additional calorimetric analysis of RNA point mutants and tc derivatives identifies functional groups crucial for the interaction and including their respective enthalpic and entropic contributions we can propose detailed structural and functional roles for certain groups. The conclusions are consistent with mutational analyses in vivo and support the hypothesis that tc binding reinforces the structure of the RNA aptamer, preventing the scanning ribosome from melting it efficiently. PMID- 16707666 TI - Role of dipicolinic acid in resistance and stability of spores of Bacillus subtilis with or without DNA-protective alpha/beta-type small acid-soluble proteins. AB - Dipicolinic acid (DPA) comprises approximately 10% of the dry weight of spores of Bacillus species. Although DPA has long been implicated in spore resistance to wet heat and spore stability, definitive evidence on the role of this abundant molecule in spore properties has generally been lacking. Bacillus subtilis strain FB122 (sleB spoVF) produced very stable spores that lacked DPA, and sporulation of this strain with DPA yielded spores with nearly normal DPA levels. DPA-replete and DPA-less FB122 spores had similar levels of the DNA protective alpha/beta type small acid-soluble spore proteins (SASP), but the DPA-less spores lacked SASP-gamma. The DPA-less FB122 spores exhibited similar UV resistance to the DPA replete spores but had lower resistance to wet heat, dry heat, hydrogen peroxide, and desiccation. Neither wet heat nor hydrogen peroxide killed the DPA-less spores by DNA damage, but desiccation did. The inability to synthesize both DPA and most alpha/beta-type SASP in strain PS3664 (sspA sspB sleB spoVF) resulted in spores that lost viability during sporulation, at least in part due to DNA damage. DPA-less PS3664 spores were more sensitive to wet heat than either DPA less FB122 spores or DPA-replete PS3664 spores, and the latter also retained viability during sporulation. These and previous results indicate that, in addition to alpha/beta-type SASP, DPA also is extremely important in spore resistance and stability and, further, that DPA has some specific role(s) in protecting spore DNA from damage. Specific roles for DPA in protecting spore DNA against damage may well have been a major driving force for the spore's accumulation of the high levels of this small molecule. PMID- 16707664 TI - Analysis of hepatitis C virus RNA dimerization and core-RNA interactions. AB - The core protein of hepatitis C virus (HCV) has been shown previously to act as a potent nucleic acid chaperone in vitro, promoting the dimerization of the 3' untranslated region (3'-UTR) of the HCV genomic RNA, a process probably mediated by a small, highly conserved palindromic RNA motif, named DLS (dimer linkage sequence) [G. Cristofari, R. Ivanyi-Nagy, C. Gabus, S. Boulant, J. P. Lavergne, F. Penin and J. L. Darlix (2004) Nucleic Acids Res., 32, 2623-2631]. To investigate in depth HCV RNA dimerization, we generated a series of point mutations in the DLS region. We find that both the plus-strand 3'-UTR and the complementary minus-strand RNA can dimerize in the presence of core protein, while mutations in the DLS (among them a single point mutation that abolished RNA replication in a HCV subgenomic replicon system) completely abrogate dimerization. Structural probing of plus- and minus-strand RNAs, in their monomeric and dimeric forms, indicate that the DLS is the major if not the sole determinant of UTR RNA dimerization. Furthermore, the N-terminal basic amino acid clusters of core protein were found to be sufficient to induce dimerization, suggesting that they retain full RNA chaperone activity. These findings may have important consequences for understanding the HCV replicative cycle and the genetic variability of the virus. PMID- 16707665 TI - A complex transcription network controls the early stages of biofilm development by Escherichia coli. PMID- 16707667 TI - Different roles of EIIABMan and EIIGlc in regulation of energy metabolism, biofilm development, and competence in Streptococcus mutans. AB - The phosphoenolpyruvate:sugar phosphotransferase system (PTS) is the major carbohydrate transport system in oral streptococci. The mannose-PTS of Streptococcus mutans, which transports mannose and glucose, is involved in carbon catabolite repression (CCR) and regulates the expression of known virulence genes. In this study, we investigated the role of EII(Glc) and EIIAB(Man) in sugar metabolism, gene regulation, biofilm formation, and competence. The results demonstrate that the inactivation of ptsG, encoding a putative EII(Glc), did not lead to major changes in sugar metabolism or affect the phenotypes of interest. However, the loss of EII(Glc) was shown to have a significant impact on the proteome and to affect the expression of a known virulence factor, fructan hydrolase (fruA). JAM1, a mutant strain lacking EIIAB(Man), had an impaired capacity to form biofilms in the presence of glucose and displayed a decreased ability to be transformed with exogenous DNA. Also, the lactose- and cellobiose PTSs were positively and negatively regulated by EIIAB(Man), respectively. Microarrays were used to investigate the profound phenotypic changes displayed by JAM1, revealing that EIIAB(Man) of S. mutans has a key regulatory role in energy metabolism, possibly by sensing the energy levels of the cells or the carbohydrate availability and, in response, regulating the activity of transcription factors and carbohydrate transporters. PMID- 16707668 TI - Isolation and characterization of VceC gain-of-function mutants that can function with the AcrAB multiple-drug-resistant efflux pump of Escherichia coli. AB - VceC is the outer membrane component of the major facilitator (MF) VceAB-VceC multiple-drug-resistant (MDR) efflux pump of Vibrio cholerae. TolC is the outer membrane component of the resistance-nodulation-division AcrAB-TolC efflux pump of Escherichia coli. Although these proteins share little amino acid sequence identity, their crystal structures can be readily superimposed upon one another. In this study, we have asked if TolC and VceC are interchangeable for the functioning of the AcrAB and VceAB pumps. We have found that TolC can replace VceC to form a functional VceAB-TolC MDR pump, but VceC cannot replace TolC to form a functional AcrAB-VceC pump. However, we have been able to isolate gain-of function (gof) VceC mutants which can functionally interface with AcrAB. These mutations map to four different amino acids located at the periplasmic tip of VceC. Chemical cross-linkage experiments indicate that both wild-type and gof mutant VceC can physically interact with the AcrAB complex, suggesting that these gof mutations are not affecting the recruitment of VceC to the AcrAB complex but rather its ability to functionally interface with the AcrAB pump. PMID- 16707669 TI - Proteomic insights into metabolic adaptations in Alcanivorax borkumensis induced by alkane utilization. AB - Alcanivorax borkumensis is a ubiquitous marine petroleum oil-degrading bacterium with an unusual physiology specialized for alkane metabolism. This "hydrocarbonoclastic" bacterium degrades an exceptionally broad range of alkane hydrocarbons but few other substrates. The proteomic analysis presented here reveals metabolic features of the hydrocarbonoclastic lifestyle. Specifically, hexadecane-grown and pyruvate-grown cells differed in the expression of 97 cytoplasmic and membrane-associated proteins whose genes appeared to be components of 46 putative operon structures. Membrane proteins up-regulated in alkane-grown cells included three enzyme systems able to convert alkanes via terminal oxidation to fatty acids, namely, enzymes encoded by the well-known alkB1 gene cluster and two new alkane hydroxylating systems, a P450 cytochrome monooxygenase and a putative flavin-binding monooxygenase, and enzymes mediating beta-oxidation of fatty acids. Cytoplasmic proteins up-regulated in hexadecane grown cells reflect a central metabolism based on a fatty acid diet, namely, enzymes of the glyoxylate bypass and of the gluconeogenesis pathway, able to provide key metabolic intermediates, like phosphoenolpyruvate, from fatty acids. They also include enzymes for synthesis of riboflavin and of unsaturated fatty acids and cardiolipin, which presumably reflect membrane restructuring required for membranes to adapt to perturbations induced by the massive influx of alkane oxidation enzymes. Ancillary functions up-regulated included the lipoprotein releasing system (Lol), presumably associated with biosurfactant release, and polyhydroxyalkanoate synthesis enzymes associated with carbon storage under conditions of carbon surfeit. The existence of three different alkane-oxidizing systems is consistent with the broad range of oil hydrocarbons degraded by A. borkumensis and its ecological success in oil-contaminated marine habitats. PMID- 16707670 TI - The ectD gene, which is involved in the synthesis of the compatible solute hydroxyectoine, is essential for thermoprotection of the halophilic bacterium Chromohalobacter salexigens. AB - The halophilic bacterium Chromohalobacter salexigens synthesizes and accumulates compatible solutes in response to salt and temperature stress. (13)C-nuclear magnetic resonance analysis of cells grown in minimal medium at the limiting temperature of 45 degrees C revealed the presence of hydroxyectoine, ectoine, glutamate, trehalose (not present in cells grown at 37 degrees C), and the ectoine precursor, Ngamma-acetyldiaminobutyric acid. High-performance liquid chromatography analyses showed that the levels of ectoine and hydroxyectoine were maximal during the stationary phase of growth. Accumulation of hydroxyectoine was up-regulated by salinity and temperature, whereas accumulation of ectoine was up regulated by salinity and down-regulated by temperature. The ectD gene, which is involved in the conversion of ectoine to hydroxyectoine, was isolated as part of a DNA region that also contains a gene whose product belongs to the AraC-XylS family of transcriptional activators. Orthologs of ectD were found within the sequenced genomes of members of the proteobacteria, firmicutes, and actinobacteria, and their products were grouped into the ectoine hydroxylase subfamily, which was shown to belong to the superfamily of Fe(II)- and 2 oxoglutarate-dependent oxygenases. Analysis of the ectoine and hydroxyectoine contents of an ectABC ectD mutant strain fed with 1 mM ectoine or hydroxyectoine demonstrated that ectD is required for the main ectoine hydroxylase activity in C. salexigens. Although in minimal medium at 37 degrees C the wild-type strain grew with 0.5 to 3.0 M NaCl, with optimal growth at 1.5 M NaCl, at 45 degrees C it could not cope with the lowest (0.75 M NaCl) or the highest (3.0 M NaCl) salinity, and it grew optimally at 2.5 M NaCl. The ectD mutation caused a growth defect at 45 degrees C in minimal medium with 1.5 to 2.5 M NaCl, but it did not affect growth at 37 degrees C at any salinity tested. With 2.5 M NaCl, the ectD mutant synthesized 38% (at 37 degrees C) and 15% (at 45 degrees C) of the hydroxyectoine produced by the wild-type strain. All of these data reveal that hydroxyectoine synthesis mediated by the ectD gene is thermoregulated and essential for thermoprotection of C. salexigens. PMID- 16707671 TI - Characterization of the siderophore of Francisella tularensis and role of fslA in siderophore production. AB - We determined that LVS and Schu S4 strains of the human pathogen Francisella tularensis express a siderophore when grown under iron-limiting conditions. We purified this siderophore by conventional column chromatography and high-pressure liquid chromatography and used mass spectrometric analysis to demonstrate that it is structurally similar to the polycarboxylate siderophore rhizoferrin. The siderophore promoted the growth of LVS and Schu S4 strains in iron-limiting media. We identified a potential siderophore biosynthetic gene cluster encoded by fslABCD in the F. tularensis genome. The first gene in the cluster, fslA, encodes a member of the superfamily of nonribosomal peptide synthetase-independent siderophore synthetases (NIS synthetases) characterized by the aerobactin synthetases IucA and IucC. We determined that fslA is transcribed as part of an operon with downstream gene fslB and that the expression of the locus is induced by iron starvation. A targeted in-frame nonpolar deletion of fslA in LVS resulted in the loss of siderophore expression and in a reduced ability of F. tularensis to grow under conditions of iron limitation. Siderophore activity and the ability to grow under iron limitation could be regained by introducing the fslA(+) gene on a complementing plasmid. Our results suggest that the fslA-dependent siderophore is important for survival of F. tularensis in an iron-deficient environment. PMID- 16707673 TI - Spatial localizations of Mam22 and Mam12 in the magnetosomes of Magnetospirillum magnetotacticum. AB - Magnetospirillum magnetotacticum possesses intracellular magnetite particles with a chain-like structure, termed magnetosomes. The bacterium expresses 22-kDa and 12-kDa magnetosome-associated proteins, termed Mam22 (MamA) and Mam12 (MamC), respectively. In this study, we investigated the structure of the purified magnetosomes with transmission electron microscopic techniques and found that the magnetosomes consisted of four compartments, i.e., magnetite crystal, magnetosomal membrane, interparticle connection, and magnetosomal matrix. Furthermore, we determined the precise localizations of Mam22 and Mam12 using immunogold staining of the purified magnetosomes and ultrathin sections of the bacterial cells. Interestingly, most Mam22 existed in the magnetosomal matrix, whereas Mam12 was strictly localized in the magnetosomal membrane. Moreover, the recombinant Mam22 was attached to the magnetosomal matrix of the Mam22-deficient magnetosomes prepared by alkaline treatment, such as 0.1 M Caps-NaOH buffer (pH 11.0). The spatial localization of the magnetosome-associated proteins in the magnetosomal chain provides useful information to elucidate the functional roles of these proteins. PMID- 16707672 TI - Inhibition of ATP hydrolysis by thermoalkaliphilic F1Fo-ATP synthase is controlled by the C terminus of the epsilon subunit. AB - The F(1)F(o)-ATP synthases of alkaliphilic bacteria exhibit latent ATPase activity, and for the thermoalkaliphile Bacillus sp. strain TA2.A1, this activity is intrinsic to the F(1) moiety. To study the mechanism of ATPase inhibition, we developed a heterologous expression system in Escherichia coli to produce TA2F(1) complexes from this thermoalkaliphile. Like the native F(1)F(o)-ATP synthase, the recombinant TA2F(1) was blocked in ATP hydrolysis activity, and this activity was stimulated by the detergent lauryldimethylamine oxide. To determine if the C terminal domain of the epsilon subunit acts as an inhibitor of ATPase activity and if an electrostatic interaction plays a role, a TA2F(1) mutant with either a truncated epsilon subunit [i.e., TA2F(1)(epsilon(DeltaC))] or substitution of basic residues in the second alpha-helix of epsilon with nonpolar alanines [i.e., TA2F(1)(epsilon(6A))] was constructed. Both mutants showed ATP hydrolysis activity at low and high concentrations of ATP. Treatment of the purified F(1)F(o)-ATP synthase and TA2F(1)(epsilon(WT)) complex with proteases revealed that the epsilon subunit was resistant to proteolytic digestion. In contrast, the epsilon subunit of TA2F(1)(epsilon(6A)) was completely degraded by trypsin, indicating that the C-terminal arm was in a conformation where it was no longer protected from proteolytic digestion. In addition, ATPase activity was not further activated by protease treatment when compared to the untreated control, supporting the observation that epsilon was responsible for inhibition of ATPase activity. To study the effect of the alanine substitutions in the epsilon subunit in the entire holoenzyme, we reconstituted recombinant TA2F(1) complexes with F(1)-stripped native membranes of strain TA2.A1. The reconstituted TA2F(o)F(1)(epsilon(WT)) was blocked in ATP hydrolysis and exhibited low levels of ATP-driven proton pumping consistent with the F(1)F(o)-ATP synthase in native membranes. Reconstituted TA2F(o)F(1)(epsilon(6A)) exhibited ATPase activity that correlated with increased ATP-driven proton pumping, confirming that the epsilon subunit also inhibits ATPase activity of TA2F(o)F(1). PMID- 16707675 TI - Ectopic overexpression of wild-type and mutant hipA genes in Escherichia coli: effects on macromolecular synthesis and persister formation. AB - Persistence is an epigenetic trait that allows a small fraction of bacteria, approximately one in a million, to survive prolonged exposure to antibiotics. In Escherichia coli an increased frequency of persisters, called "high persistence," is conferred by mutations in the hipA gene, which encodes the toxin entity of the toxin-antitoxin module hipBA. The high-persistence allele hipA7 was originally identified because of its ability to confer high persistence, but little is known about the physiological role of the wild-type hipA gene. We report here that the expression of wild-type hipA in excess of hipB inhibits protein, RNA, and DNA synthesis in vivo. However, unlike the RelE and MazF toxins, HipA had no effect on protein synthesis in an in vitro translation system. Moreover, the expression of wild-type hipA conferred a transient dormant state (persistence) to a sizable fraction of cells, whereas the rest of the cells remained in a prolonged dormant state that, under appropriate conditions, could be fully reversed by expression of the cognate antitoxin gene hipB. In contrast, expression of the mutant hipA7 gene in excess of hipB did not markedly inhibit protein synthesis as did wild type hipA and yet still conferred persistence to ca. 10% of cells. We propose that wild-type HipA, upon release from HipB, is able to inhibit macromolecular synthesis and induces a bacteriostatic state that can be reversed by expression of the hipB gene. However, the ability of the wild-type hipA gene to generate a high frequency of persisters, equal to that conferred by the hipA7 allele, may be distinct from the ability to block macromolecular synthesis. PMID- 16707674 TI - Functional analysis of glucan binding protein B from Streptococcus mutans. AB - Mutans streptococci are major etiological agents of dental caries, and several of their secreted products contribute to bacterial accumulation on teeth. Of these, Streptococcus mutans glucan binding protein B (GbpB) is a novel, immunologically dominant protein. Its biological function is unclear, although GbpB shares homology with a putative peptidoglycan hydrolase from S. agalactiae and S. pneumoniae, indicative of a role in murein biosynthesis. To determine the cellular function of GbpB, we used several approaches to inactivate the gene, analyze its expression, and identify interacting proteins. None of the transformants analyzed were true gbpB mutants, since they all contained both disrupted and wild-type gene copies, and expression of functional GbpB was always conserved. Thus, the inability to obtain viable gbpB null mutants supports the notion that gbpB is an essential gene. Northern blot and real-time PCR analyses suggested that induction of gbpB expression in response to stress was a strain dependent phenomenon. Proteins that interacted with GbpB were identified in pull down and coimmunoprecipitation assays, and these data suggest that GbpB interacts with ribosomal protein L7/L12, possibly as part of a protein complex involved in peptidoglycan synthesis and cell division. PMID- 16707676 TI - The superoxide dismutases of Bacillus anthracis do not cooperatively protect against endogenous superoxide stress. AB - The Bacillus anthracis chromosome encodes four unique, putative superoxide dismutase (sod) genes. During exponential growth and sporulation, sodA1, sodA2, and sodC are transcribed constitutively throughout the growth cycle as individual genes. In contrast, the transcription of sod15 occurs mainly during late exponential and sporulation phases as part of a four-gene operon that may be involved in spore formation. Vegetative cell and spore lysates of wild-type Sterne and superoxide dismutase deletion (Deltasod) mutants show detectable SOD activity for SODA1 and SODA2, and protein analysis suggests that these two proteins form active homodimers and heterodimers. A comparison of the growth of parental versus Deltasod mutants under various chemical oxidative stresses indicates that DeltasodA1 mutants are particularly sensitive to endogenously produced superoxide, whereas DeltasodA2, Deltasod15, and DeltasodC mutants remain as resistant to this stress as the parental strain. In addition, in mouse survival assays, Deltasod15 and DeltasodA1 were responsible for less end-point death, but the level of decreased virulence does not fall within a statistically significant range. Collectively, these data show that sodA1 acts as a major protectant from intracellular superoxide stress, that sod15 is transcribed as part of an operon that may play a role in cell morphology, and that sodA2 and sodC may have minor roles that are not apparent in the conditions tested here. PMID- 16707677 TI - Complete cellulase system in the marine bacterium Saccharophagus degradans strain 2-40T. AB - Saccharophagus degradans strain 2-40 is a representative of an emerging group of marine complex polysaccharide (CP)-degrading bacteria. It is unique in its metabolic versatility, being able to degrade at least 10 distinct CPs from diverse algal, plant and invertebrate sources. The S. degradans genome has been sequenced to completion, and more than 180 open reading frames have been identified that encode carbohydrases. Over half of these are likely to act on plant cell wall polymers. In fact, there appears to be a full array of enzymes that degrade and metabolize plant cell walls. Genomic and proteomic analyses reveal 13 cellulose depolymerases complemented by seven accessory enzymes, including two cellodextrinases, three cellobiases, a cellodextrin phosphorylase, and a cellobiose phosphorylase. Most of these enzymes exhibit modular architecture, and some contain novel combinations of catalytic and/or substrate binding modules. This is exemplified by endoglucanase Cel5A, which has three internal family 6 carbohydrate binding modules (CBM6) and two catalytic modules from family five of glycosyl hydrolases (GH5) and by Cel6A, a nonreducing-end cellobiohydrolase from family GH6 with tandem CBM2s. This is the first report of a complete and functional cellulase system in a marine bacterium with a sequenced genome. PMID- 16707678 TI - Metabolism of isovanillate, vanillate, and veratrate by Comamonas testosteroni strain BR6020. AB - In Comamonas testosteroni strain BR6020, metabolism of isovanillate (iVan; 3 hydroxy-4-methoxybenzoate), vanillate (Van; 4-hydroxy-3-methoxybenzoate), and veratrate (Ver; 3,4-dimethoxybenzoate) proceeds via protocatechuate (Pca; 3,4 dihydroxybenzoate). A 13.4-kb locus coding for the catabolic enzymes that channel the three substrates to Pca was cloned. O demethylation is mediated by the phthalate family oxygenases IvaA (converts iVan to Pca and Ver to Van) and VanA (converts Van to Pca and Ver to iVan). Reducing equivalents from NAD(P)H are transferred to the oxygenases by the class IA oxidoreductase IvaB. Studies using whole cells, cell extracts, and reverse transcriptase PCR showed that degradative activity and expression of vanA, ivaA, and ivaB are inducible. In succinate- and Pca-grown cells, there is negligible degradative activity towards Van, Ver, and iVan and little to no expression of vanA, ivaA, and ivaB. Growth on Van or Ver results in production of oxygenases with activity towards Van, Ver, and iVan and expression of vanA, ivaA, and ivaB. With iVan-grown cultures, ivaA and ivaB are expressed, and in assays with whole cells, production of the iVan oxygenase is observed, but there is little activity towards Van or Ver. In cell extracts, though, Ver metabolism is observed, which suggests that the system mediating iVan uptake in whole cells does not mediate Ver uptake. PMID- 16707680 TI - Albomycin uptake via a ferric hydroxamate transport system of Streptococcus pneumoniae R6. AB - The antibiotic albomycin is highly effective against Streptococcus pneumoniae, with an MIC of 10 ng/ml. The reason for the high efficacy was studied by measuring the uptake of albomycin into S. pneumoniae. Albomycin was transported via the system that transports the ferric hydroxamates ferrichrome and ferrioxamine B. These two ferric hydroxamates antagonized the growth inhibition by albomycin and salmycin. Cross-inhibition of the structurally different ferric hydroxamates to both antibiotics can be explained by the similar iron coordination centers of the four compounds. [(55)Fe(3+)]ferrichrome and [(55)Fe(3+)]ferrioxamine B were taken up by the same transport system into S. pneumoniae. Mutants in the adjacent fhuD, fhuB, and fhuG genes were transport inactive and resistant to the antibiotics. Albomycin, ferrichrome, ferrioxamine B, and salmycin bound to the isolated FhuD protein and prevented degradation by proteinase K. The fhu locus consisting of the fhuD, fhuB, fhuG, and fhuC genes determines a predicted ABC transporter composed of the FhuD binding lipoprotein, the FhuB and FhuG transport proteins, and the FhuC ATPase. It is concluded that active transport of albomycin mediates the high antibiotic efficacy in S. pneumoniae. PMID- 16707679 TI - The Haemophilus influenzae Type b hcsA and hcsB gene products facilitate transport of capsular polysaccharide across the outer membrane and are essential for virulence. AB - Haemophilus influenzae type b is a common cause of invasive bacterial disease, especially among children in underdeveloped countries. The type b polysaccharide capsule is a polymer of ribose and ribitol-5-phosphate and is a critical determinant of virulence. Expression of the type b capsule is dependent upon the cap b locus, which consists of three functionally distinct regions, designated regions 1 to 3. Region 3 contains the hcsA and hcsB genes, which share significant homology with genes that have been implicated in encapsulation in other pathogenic bacteria but have unclear functions. In this study, we inactivated hcsA alone, hcsB alone, and both hcsA and hcsB together and examined the effects of these mutations on polysaccharide transport and bacterial virulence properties. Inactivation of hcsA alone resulted in accumulation of polysaccharide in the periplasm and a partial decrease in surface-associated polysaccharide, whereas inactivation of hcsB alone or of both hcsA and hcsB together resulted in accumulation of polysaccharide in the periplasm and complete loss of surface-associated polysaccharide. All mutations eliminated serum resistance and abrogated bacteremia and mortality in neonatal rats. These results indicate that the hcsA and hcsB gene products have complementary functions involved in the transport of polysaccharide across the outer membrane and are essential for virulence. PMID- 16707681 TI - Overexpression of PrfA leads to growth inhibition of Listeria monocytogenes in glucose-containing culture media by interfering with glucose uptake. AB - Listeria monocytogenes strains expressing high levels of the virulence regulator PrfA (mutant PrfA* or wild-type PrfA) show strong growth inhibition in minimal media when they are supplemented with glucose but not when they are supplemented with glucose-6-phosphate compared to the growth of isogenic strains expressing low levels of PrfA. A significantly reduced rate of glucose uptake was observed in a PrfA*-overexpressing strain growing in LB supplemented with glucose. Comparative transcriptome analyses were performed with RNA isolated from a prfA mutant and an isogenic strain carrying multiple copies of prfA or prfA* on a plasmid. These analyses revealed that in addition to high transcriptional up regulation of the known PrfA-regulated virulence genes (group I), there was less pronounced up-regulation of the expression of several phage and metabolic genes (group II) and there was strong down-regulation of several genes involved mainly in carbon and nitrogen metabolism in the PrfA*-overexpressing strain (group III). Among the latter genes are the nrgAB, gltAB, and glnRA operons (involved in nitrogen metabolism), the ilvB operon (involved in biosynthesis of the branched chain amino acids), and genes for some ABC transporters. Most of the down regulated genes have been shown previously to belong to a class of genes in Bacillus subtilis whose expression is negatively affected by impaired glucose uptake. Our results lead to the conclusion that excess PrfA (or PrfA*) interferes with a component(s) essential for phosphotransferase system-mediated glucose transport. PMID- 16707682 TI - Escherichia coli competence gene homologs are essential for competitive fitness and the use of DNA as a nutrient. AB - Natural genetic competence is the ability of cells to take up extracellular DNA and is an important mechanism for horizontal gene transfer. Another potential benefit of natural competence is that exogenous DNA can serve as a nutrient source for starving bacteria because the ability to "eat" DNA is necessary for competitive survival in environments containing limited nutrients. We show here that eight Escherichia coli genes, identified as homologs of com genes in Haemophilus influenzae and Neisseria gonorrhoeae, are necessary for the use of extracellular DNA as the sole source of carbon and energy. These genes also confer a competitive advantage to E. coli during long-term stationary-phase incubation. We also show that homologs of these genes are found throughout the proteobacteria, suggesting that the use of DNA as a nutrient may be a widespread phenomenon. PMID- 16707683 TI - Maltose and maltodextrin utilization by Bacillus subtilis. AB - Bacillus subtilis can utilize maltose and maltodextrins that are derived from polysaccharides, like starch or glycogen. In this work, we show that maltose is taken up by a member of the phosphoenolpyruvate-dependent phosphotransferase system and maltodextrins are taken up by a maltodextrin-specific ABC transporter. Uptake of maltose by the phosphoenolpyruvate-dependent phosphotransferase system is mediated by maltose-specific enzyme IICB (MalP; synonym, GlvC), with an apparent K(m) of 5 microM and a V(max) of 91 nmol . min(-1) . (10(10) CFU)(-1). The maltodextrin-specific ABC transporter is composed of the maltodextrin binding protein MdxE (formerly YvdG), with affinities in the low micromolar range for maltodextrins, and the membrane-spanning components MdxF and MdxG (formerly YvdH and YvdI, respectively), as well as the energizing ATPase MsmX. Maltotriose transport occurs with an apparent K(m) of 1.4 microM and a V(max) of 4.7 nmol . min(-1) . (10(10) CFU)(-1). PMID- 16707684 TI - Evolution of immunity and host chromosome integration site of P2-like coliphages. AB - The amount and distribution of variation in the genomic region containing the genes in the lytic-lysogenic genetic switch and the sequence that determines the integration site into the host chromosome were analyzed for 38 P2-like phages from Escherichia coli. The genetic switch consists of two convergent mutually exclusive promoters, Pe and Pc, and two repressors, C and Cox. The immunity repressor C blocks the early Pe promoter, leading to the establishment of lysogeny. The Cox repressor blocks expression of Pc, allowing lytic growth. Phylogenetic analyses showed that the C and Cox proteins were distributed into seven distinct classes. The phylogenetic relationship differed between the two proteins, and we showed that homologous recombination plays a major role in creating alterations in the genetic switch, leading to new immunity classes. Analyses of the host integration site for these phages resulted in the discovery of a previously unknown site, and there were at least four regular integration sites. Interestingly, we found no case where phages of the same immunity class had different host attachment sites. The evolution of immunity and integration sites is complex, since it involves interactions both between the phages themselves and between phages and hosts, and often, both regulatory proteins and target DNA must change. PMID- 16707685 TI - 6S RNA regulation of pspF transcription leads to altered cell survival at high pH. AB - 6S RNA is a highly abundant small RNA that regulates transcription through direct interaction with RNA polymerase. Here we show that 6S RNA directly inhibits transcription of pspF, which subsequently leads to inhibition of pspABCDE and pspG expression. Cells without 6S RNA are able to survive at elevated pH better than wild-type cells due to loss of 6S RNA-regulation of pspF. This 6S RNA dependent phenotype is eliminated in pspF-null cells, indicating that 6S RNA effects are conferred through PspF. Similar growth phenotypes are seen when PspF levels are increased in a 6S RNA-independent manner, signifying that changes to pspF expression are sufficient. Changes in survival at elevated pH most likely result from altered expression of pspABCDE and/or pspG, both of which require PspF for transcription and are indirectly regulated by 6S RNA. 6S RNA provides another layer of regulation in response to high pH during stationary phase. We propose that the normal role of 6S RNA at elevated pH is to limit the extent of the psp response under conditions of nutrient deprivation, perhaps facilitating appropriate allocation of diminishing resources. PMID- 16707686 TI - Mutationally altered signal output in the Nart (NarX-Tar) hybrid chemoreceptor. AB - Signal-transducing proteins that span the cytoplasmic membrane transmit information about the environment to the interior of the cell. In bacteria, these signal transducers include sensor kinases, which typically control gene expression via response regulators, and methyl-accepting chemoreceptor proteins, which control flagellar rotation via the CheA kinase and CheY response regulator. We previously reported that a chimeric protein (Nart) that joins the ligand binding, transmembrane, and linker regions of the NarX sensor kinase to the signaling and adaptation domains of the Tar chemoreceptor elicits a repellent response to nitrate and nitrite. As with NarX, nitrate evokes a stronger response than nitrite. Here we show that mutations targeting a highly conserved sequence (the P box) in the periplasmic domain alter chemoreception by Nart and signaling by NarX similarly. In particular, the G51R substitution converts Nart from a repellent receptor into an attractant receptor for nitrate. Our results underscore the conclusion that the fundamental mechanism of transmembrane signaling is conserved between homodimeric sensor kinases and chemoreceptors. They also highlight the plasticity of the coupling between ligand binding and signal output in these systems. PMID- 16707687 TI - Bundle-forming pili and EspA are involved in biofilm formation by enteropathogenic Escherichia coli. AB - Microcolony formation is one of the initial steps in biofilm development, and in enteropathogenic Escherichia coli (EPEC) it is mediated by several adhesins, including the bundle-forming pilus (BFP) and the EspA filament. Here we report that EPEC forms biofilms on plastic under static conditions and a flowthrough continuous culture system. The abilities of several EPEC isogenic mutants to form biofilms were assessed. Adhesins such as BFP and EspA, important in microcolony formation on epithelial cells, are also involved in bacterial aggregation during biofilm formation on abiotic surfaces. Mutants that do not express BFP or EspA form more-diffuse biofilms than does the wild type. We also determined, using gfp transcriptional fusions, that, consistent with the role of these adhesins in biofilms, the genes encoding BFP and EspA are expressed during biofilm formation. Finally, expression of espA is controlled by a quorum-sensing (QS) regulatory mechanism, and the EPEC qseA QS mutant also forms altered biofilms, suggesting that this signaling mechanism plays an important role in EPEC biofilm development. Taken together, these studies allowed us to propose a model of EPEC biofilm formation. PMID- 16707688 TI - Differential activation of Escherichia coli chemoreceptors by blue-light stimuli. AB - Enteric bacteria tumble, swim slowly, and are then paralyzed upon exposure to 390 to 530-nm light. Here, we analyze this complex response in Escherichia coli using standard fluorescence microscope optics for excitation at 440 +/- 5 nm. The slow swimming and paralysis occurred only in dye-containing growth media or buffers. Excitation elicited complete paralysis within a second in 1 muM proflavine dye, implying specific motor damage, but prolonged tumbling in buffer alone. The tumbling half-response times were subsecond for onset but more than a minute for recovery. The response required the chemotaxis signal protein CheY and receptor-dependent activation of its kinase CheA. The study of deletion mutants revealed a specific requirement for either the aerotaxis receptor Aer or the chemoreceptor Tar but not the Tar homolog Tsr. The action spectrum of the wild type response was consistent with a flavin, but the chromophores remain to be identified. The motile response processed via Aer was sustained, with recovery to either step-up or -down taking more than a minute. The response processed via Tar was transient, recovering on second time scales comparable to chemotactic responses. The response duration and amplitude were dependent on relative expression of Aer, Tar, and Tsr. The main response features were reproduced when each receptor was expressed singly from a plasmid in a receptorless host strain. However, time-resolved motion analysis revealed subtle kinetic differences that reflect the role of receptor cluster interactions in kinase activation deactivation dynamics. PMID- 16707689 TI - Anatomy of a lactococcal phage tail. AB - Bacteriophages of the Siphoviridae family utilize a long noncontractile tail to recognize, adsorb to, and inject DNA into their bacterial host. The tail anatomy of the archetypal Siphoviridae lambda has been well studied, in contrast to phages infecting gram-positive bacteria. This report outlines a detailed anatomical description of a typical member of the Siphoviridae infecting a gram positive bacterium. The tail superstructure of the lactococcal phage Tuc2009 was investigated using N-terminal protein sequencing, Western blotting, and immunogold transmission electron microscopy, allowing a tangible path to be followed from gene sequence through encoded protein to specific architectural structures on the Tuc2009 virion. This phage displays a striking parity with lambda with respect to tail structure, which reenforced a model proposed for Tuc2009 tail architecture. Furthermore, comparisons with lambda and other lactococcal phages allowed the specification of a number of genetic submodules likely to encode specific tail structures. PMID- 16707690 TI - Crl activates transcription initiation of RpoS-regulated genes involved in the multicellular behavior of Salmonella enterica serovar Typhimurium. AB - In Salmonella enterica serovar Typhimurium, the stationary-phase sigma factor sigma(S) (RpoS) is required for virulence, stress resistance, biofilm formation, and development of the rdar morphotype. This morphotype is a multicellular behavior characterized by expression of the adhesive extracellular matrix components cellulose and curli fimbriae. The Crl protein of Escherichia coli interacts with sigma(S) and activates expression of sigma(S)-regulated genes, such as the csgBAC operon encoding the subunit of the curli proteins, by an unknown mechanism. Here, we showed using in vivo and in vitro experiments that the Crl protein of Salmonella serovar Typhimurium is required for development of a typical rdar morphotype and for maximal expression of the csgD, csgB, adrA, and bcsA genes, which are involved in curli and cellulose biosynthesis. In vitro transcription assays and potassium permanganate reactivity experiments with purified His(6)-Crl showed that Crl directly activated sigma(S)-dependent transcription initiation at the csgD and adrA promoters. We observed no effect of Crl on sigma(70)-dependent transcription. Crl protein levels increased during the late exponential and stationary growth phases in Luria-Beratani medium without NaCl at 28 degrees C. We obtained complementation of the crl mutation by increasing sigma(S) levels. This suggests that Crl has a major physiological impact at low concentrations of sigma(S). PMID- 16707691 TI - Contribution of the PhoP-PhoQ and PmrA-PmrB two-component regulatory systems to Mg2+-induced gene regulation in Pseudomonas aeruginosa. AB - When grown in divalent cation-limited medium, Pseudomonas aeruginosa becomes resistant to cationic antimicrobial peptides and polymyxin B. This resistance is regulated by the PhoP-PhoQ and PmrA-PmrB two-component regulatory systems. To further characterize Mg(2+) regulation in P. aeruginosa, microarray transcriptional profiling was conducted to compare wild-type P. aeruginosa grown under Mg(2+)-limited and Mg(2+)-replete conditions to isogenic phoP and pmrA mutants grown under Mg(2+)-limited conditions. Under Mg(2+)-limited conditions (0.02 mM Mg(2+)), approximately 3% of the P. aeruginosa genes were differentially expressed compared to the expression in bacteria grown under Mg(2+)-replete conditions (2 mM Mg(2+)). Only a modest subset of the Mg(2+)-regulated genes were regulated through either PhoP or PmrA. To determine which genes were directly regulated, a bioinformatic search for conserved binding motifs was combined with confirmatory reverse transcriptase PCR and gel shift promoter binding assays, and the results indicated that very few genes were directly regulated by these response regulators. It was found that in addition to the previously known oprH phoP-phoQ operon and the pmrHFIJKLM-ugd operon, the PA0921 and PA1343 genes, encoding small basic proteins, were regulated by Mg(2+) in a PhoP-dependent manner. The number of known PmrA-regulated genes was expanded to include the PA1559-PA1560, PA4782-PA4781, and feoAB operons, in addition to the previously known PA4773-PA4775-pmrAB and pmrHFIJKLM-ugd operons. PMID- 16707692 TI - Active transcription of rRNA operons is a driving force for the distribution of RNA polymerase in bacteria: effect of extrachromosomal copies of rrnB on the in vivo localization of RNA polymerase. AB - In contrast to eukaryotes, bacteria such as Escherichia coli contain only one form of RNA polymerase (RNAP), which is responsible for all cellular transcription. Using an RNAP-green fluorescent protein fusion protein, we showed previously that E. coli RNAP is partitioned exclusively in the nucleoid and that stable RNA synthesis, particularly rRNA transcription, is critical for concentrating a significant fraction of RNAP in transcription foci during exponential growth. The extent of focus formation varies under different physiological conditions, supporting the proposition that RNAP redistribution is an important element for global gene regulation. Here we show that extra, plasmid borne copies of an rRNA operon recruit RNAP from the nucleoid into the cytoplasmic space and that this is accompanied by a reduction in the growth rate. Transcription of an intact rRNA operon is not necessary, although a minimal transcript length is required for this phenotype. Replacement of the ribosomal promoters with another strong promoter, Ptac, abolished the effect. These results demonstrate that active synthesis from rRNA promoters is a major driving force for the distribution of RNAP in bacteria. The implications of our results for the regulation of rRNA synthesis and cell growth are discussed. PMID- 16707693 TI - The genome of the obligately intracellular bacterium Ehrlichia canis reveals themes of complex membrane structure and immune evasion strategies. AB - Ehrlichia canis, a small obligately intracellular, tick-transmitted, gram negative, alpha-proteobacterium, is the primary etiologic agent of globally distributed canine monocytic ehrlichiosis. Complete genome sequencing revealed that the E. canis genome consists of a single circular chromosome of 1,315,030 bp predicted to encode 925 proteins, 40 stable RNA species, 17 putative pseudogenes, and a substantial proportion of noncoding sequence (27%). Interesting genome features include a large set of proteins with transmembrane helices and/or signal sequences and a unique serine-threonine bias associated with the potential for O glycosylation that was prominent in proteins associated with pathogen-host interactions. Furthermore, two paralogous protein families associated with immune evasion were identified, one of which contains poly(G-C) tracts, suggesting that they may play a role in phase variation and facilitation of persistent infections. Genes associated with pathogen-host interactions were identified, including a small group encoding proteins (n = 12) with tandem repeats and another group encoding proteins with eukaryote-like ankyrin domains (n = 7). PMID- 16707694 TI - Structural and functional characterization of three polyketide synthase gene clusters in Bacillus amyloliquefaciens FZB 42. AB - Although bacterial polyketides are of considerable biomedical interest, the molecular biology of polyketide biosynthesis in Bacillus spp., one of the richest bacterial sources of bioactive natural products, remains largely unexplored. Here we assign for the first time complete polyketide synthase (PKS) gene clusters to Bacillus antibiotics. Three giant modular PKS systems of the trans acyltransferase type were identified in Bacillus amyloliquefaciens FZB 42. One of them, pks1, is an ortholog of the pksX operon with a previously unknown function in the sequenced model strain Bacillus subtilis 168, while the pks2 and pks3 clusters are novel gene clusters. Cassette mutagenesis combined with advanced mass spectrometric techniques such as matrix-assisted laser desorption ionization time of flight mass spectrometry and liquid chromatography-electrospray ionization mass spectrometry revealed that the pks1 (bae) and pks3 (dif) gene clusters encode the biosynthesis of the polyene antibiotics bacillaene and difficidin or oxydifficidin, respectively. In addition, B. subtilis OKB105 (pheA sfp(0)), a transformant of the B. subtilis 168 derivative JH642, was shown to produce bacillaene, demonstrating that the pksX gene cluster directs the synthesis of that polyketide. The GenBank accession numbers for gene clusters pks1(bae), pks2, and pks3(dif) are AJ 634060.2, AJ 6340601.2, and AJ 6340602.2, respectively. PMID- 16707696 TI - The X-ray structure of the haloalcohol dehalogenase HheA from Arthrobacter sp. strain AD2: insight into enantioselectivity and halide binding in the haloalcohol dehalogenase family. AB - Haloalcohol dehalogenases are bacterial enzymes that cleave the carbon-halogen bond in short aliphatic vicinal haloalcohols, like 1-chloro-2,3-propanediol, some of which are recalcitrant environmental pollutants. They use a conserved Ser-Tyr Arg catalytic triad to deprotonate the haloalcohol oxygen, which attacks the halogen-bearing carbon atom, producing an epoxide and a halide ion. Here, we present the X-ray structure of the haloalcohol dehalogenase HheA(AD2) from Arthrobacter sp. strain AD2 at 2.0-A resolution. Comparison with the previously reported structure of the 34% identical enantioselective haloalcohol dehalogenase HheC from Agrobacterium radiobacter AD1 shows that HheA(AD2) has a similar quaternary and tertiary structure but a much more open substrate-binding pocket. Docking experiments reveal that HheA(AD2) can bind both enantiomers of the haloalcohol substrate 1-p-nitrophenyl-2-chloroethanol in a productive way, which explains the low enantiopreference of HheA(AD2). Other differences are found in the halide-binding site, where the side chain amino group of Asn182 is in a position to stabilize the halogen atom or halide ion in HheA(AD2), in contrast to HheC, where a water molecule has taken over this role. These results broaden the insight into the structural determinants that govern reactivity and selectivity in the haloalcohol dehalogenase family. PMID- 16707695 TI - Acquisition and evolution of the exoU locus in Pseudomonas aeruginosa. AB - ExoU is a potent Pseudomonas aeruginosa cytotoxin translocated into host cells by the type III secretion system. A comparison of genomes of various P. aeruginosa strains showed that that the ExoU determinant is found in the same polymorphic region of the chromosome near a tRNA(Lys) gene, suggesting that exoU is a horizontally acquired virulence determinant. We used yeast recombinational cloning to characterize four distinct ExoU-encoding DNA segments. We then sequenced and annotated three of these four genomic regions. The sequence of the largest DNA segment, named ExoU island A, revealed many plasmid- and genomic island-associated genes, most of which have been conserved across a broad set of beta- and gamma-Proteobacteria. Comparison of the sequenced ExoU-encoding genomic islands to the corresponding PAO1 tRNA(Lys)-linked genomic island, the pathogenicity islands of strain PA14, and pKLC102 of clone C strains allowed us to propose a mechanism for the origin and transmission of the ExoU determinant. The evolutionary history very likely involved transposition of the ExoU determinant onto a transmissible plasmid, followed by transfer of the plasmid into different P. aeruginosa strains. The plasmid subsequently integrated into a tRNA(Lys) gene in the chromosome of each recipient, where it acquired insertion sequences and underwent deletions and rearrangements. We have also applied yeast recombinational cloning to facilitate a targeted mutagenesis of ExoU island A, further demonstrating the utility of the specific features of the yeast capture vector for functional analyses of genes on large horizontally acquired genetic elements. PMID- 16707698 TI - Salt stress in Desulfovibrio vulgaris Hildenborough: an integrated genomics approach. AB - The ability of Desulfovibrio vulgaris Hildenborough to reduce, and therefore contain, toxic and radioactive metal waste has made all factors that affect the physiology of this organism of great interest. Increased salinity is an important and frequent fluctuation faced by D. vulgaris in its natural habitat. In liquid culture, exposure to excess salt resulted in striking elongation of D. vulgaris cells. Using data from transcriptomics, proteomics, metabolite assays, phospholipid fatty acid profiling, and electron microscopy, we used a systems approach to explore the effects of excess NaCl on D. vulgaris. In this study we demonstrated that import of osmoprotectants, such as glycine betaine and ectoine, is the primary mechanism used by D. vulgaris to counter hyperionic stress. Several efflux systems were also highly up-regulated, as was the ATP synthesis pathway. Increases in the levels of both RNA and DNA helicases suggested that salt stress affected the stability of nucleic acid base pairing. An overall increase in the level of branched fatty acids indicated that there were changes in cell wall fluidity. The immediate response to salt stress included up regulation of chemotaxis genes, although flagellar biosynthesis was down regulated. Other down-regulated systems included lactate uptake permeases and ABC transport systems. The results of an extensive NaCl stress analysis were compared with microarray data from a KCl stress analysis, and unlike many other bacteria, D. vulgaris responded similarly to the two stresses. Integration of data from multiple methods allowed us to develop a conceptual model for the salt stress response in D. vulgaris that can be compared to those in other microorganisms. PMID- 16707697 TI - Genomic and functional analysis of the IncP-9 naphthalene-catabolic plasmid NAH7 and its transposon Tn4655 suggests catabolic gene spread by a tyrosine recombinase. AB - The naphthalene-catabolic (nah) genes on the incompatibility group P-9 (IncP-9) self-transmissible plasmid NAH7 from Pseudomonas putida G7 are some of the most extensively characterized genetic determinants for bacterial aerobic catabolism of aromatic hydrocarbons. In contrast to the detailed studies of its catabolic cascade and enzymatic functions, the biological characteristics of plasmid NAH7 have remained unclear. Our sequence determination in this study together with the previously deposited sequences revealed the entire structure of NAH7 (82,232 bp). Comparison of NAH7 with two other completely sequenced IncP-9 catabolic plasmids, pDTG1 and pWW0, revealed that the three plasmids share very high nucleotide similarities in a 39-kb region encoding the basic plasmid functions (the IncP-9 backbone). The backbone of NAH7 is phylogenetically more related to that of pDTG1 than that of pWW0. These three plasmids carry their catabolic gene clusters at different positions on the IncP-9 backbone. All of the NAH7-specified nah genes are located on a class II transposon, Tn4655. Our analysis of the Tn4655-encoded site-specific recombination system revealed that (i) a novel tyrosine recombinase, TnpI, catalyzed both the intra- and intermolecular recombination between two copies of the attI site, (ii) the functional attI site was located within a 119-bp segment, and (iii) the site-specific strand exchange occurred within a 30-bp segment in the 41-bp CORE site. Our results and the sequence data of other naphthalene-catabolic plasmids, pDTG1 and pND6-1, suggest a potential role of the TnpI-attI recombination system in the establishment of these catabolic plasmids. PMID- 16707700 TI - Identification of methylation sites in Thermotoga maritima chemotaxis receptors. AB - Adaptation in bacterial chemotaxis involves reversible methylation of specific glutamate residues within the cytoplasmic domains of methyl-accepting chemotaxis proteins. The specific sites of methylation in Salmonella enterica and Escherichia coli chemoreceptors, identified 2 decades ago, established a consensus sequence for methylation by methyltransferase CheR. Here we report the in vitro methylation of chemoreceptors from Thermotoga maritima, a hyperthermophile that has served as a useful source of chemotaxis proteins for structural analysis. Sites of methylation have been identified by liquid chromatography-mass spectrometry/mass spectrometry. Fifteen sites of methylation were identified within the cytoplasmic domains of four different T. maritima chemoreceptors. The results establish a consensus sequence for chemoreceptor methylation sites in T. maritima that is distinct from the previously identified consensus sequence for E. coli and S. enterica. These findings suggest that consensus sequences for posttranslational modifications in one organism may not be directly extrapolated to analogous modifications in other bacteria. PMID- 16707699 TI - Functional genomics of stress response in Pseudomonas putida KT2440. AB - The metabolically versatile soil bacterium Pseudomonas putida has to cope with numerous abiotic stresses in its habitats. The stress responses of P. putida KT2440 to 4 degrees C, pH 4.5, 0.8 M urea, and 45 mM sodium benzoate were analyzed by determining the global mRNA expression profiles and screening for stress-intolerant nonauxotrophic Tn5 transposon mutants. In 392 regulated genes or operons, 36 gene regions were differentially expressed by more than 2.5-fold, and 32 genes in 23 operons were found to be indispensable for growth during exposure to one of the abiotic stresses. The transcriptomes of the responses to urea, benzoate, and 4 degrees C correlated positively with each other but negatively with the transcriptome of the mineral acid response. The CbrAB sensor kinase, the cysteine synthase CysM, PcnB and VacB, which control mRNA stability, and BipA, which exerts transcript-specific translational control, were essential to cope with cold stress. The cyo operon was required to cope with acid stress. A functional PhoP, PtsP, RelA/SpoT modulon, and adhesion protein LapA were necessary for growth in the presence of urea, and the outer membrane proteins OmlA and FepA and the phosphate transporter PstBACS were indispensable for growth in the presence of benzoate. A lipid A acyltransferase (PP0063) was a mandatory component of the stress responses to cold, mineral acid, and benzoate. Adaptation of the membrane barrier, uptake of phosphate, maintenance of the intracellular pH and redox status, and translational control of metabolism are key mechanisms of the response of P. putida to abiotic stresses. PMID- 16707702 TI - The periplasmic folding of a cysteineless autotransporter passenger domain interferes with its outer membrane translocation. AB - Autotransporters are single polypeptides consisting of an outer membrane translocation domain mediating the translocation of a passenger domain. The periplasmic folding state of the passenger domain is controversial. By comparisons of passenger domains differing in their folding properties, our results suggest that periplasmic folding of passenger domains interferes with translocation. PMID- 16707701 TI - Subcellular partitioning of transcription factors in Bacillus subtilis. AB - RNA polymerase (RNAP) requires the interaction of various transcription elongation factors to efficiently transcribe RNA. During transcription of rRNA operons, RNAP forms highly processive antitermination complexes by interacting with NusA, NusB, NusG, NusE, and possibly several unidentified factors to increase elongation rates to around twice those observed for mRNA. In previous work we used cytological assays with Bacillus subtilis to identify the major sites of rRNA synthesis within the cell, which are called transcription foci. Using this cytological assay, in conjunction with both quantitative native polyacrylamide gel electrophoresis and Western blotting, we investigated the total protein levels and the ratios of NusB and NusG to RNAP in both antitermination and mRNA transcription complexes. We determined that the ratio of RNAP to NusG was 1:1 in both antitermination and mRNA transcription complexes, suggesting that NusG plays important regulatory roles in both complexes. A ratio of NusB to RNAP of 1:1 was calculated for antitermination complexes with just a 0.3:1 ratio in mRNA complexes, suggesting that NusB is restricted to antitermination complexes. We also investigated the cellular abundance and subcellular localization of transcription restart factor GreA. We found no evidence which suggests that GreA is involved in antitermination complex formation and that it has a cellular abundance which is around twice that of RNAP. Surprisingly, we found that the vast majority of GreA is associated with RNAP, suggesting that there is more than one binding site for GreA on RNAP. These results indicate that transcription elongation complexes are highly dynamic and are differentially segregated within the nucleoid according to their functions. PMID- 16707703 TI - Rapid succession within the Veillonella population of a developing human oral biofilm in situ. AB - Streptococci are the primary component of the multispecies oral biofilm known as supragingival dental plaque; they grow by fermentation of sugars to organic acids, e.g., lactic acid. Veillonellae, a ubiquitous component of early plaque, are unable to use sugars; they ferment organic acids, such as lactate, to a mixture of shorter-chain-length acids, CO(2), and hydrogen. Certain veillonellae bind to (coaggregate with) streptococci in vitro. We show that, between 4 and 8 hours into plaque development, the dominant strains of Veillonella change in their phenotypic characteristics (coaggregation and antibody reactivity) as well as in their genotypic characteristics (16S RNA gene sequences as well as strain level fingerprint patterns). This succession is coordinated with the development of mixed-species bacterial colonies. Changes in community structure can occur very rapidly in natural biofilm development, and we suggest that this process may influence evolution within this ecosystem. PMID- 16707704 TI - Molecular basis of L-rhamnose branch formation in streptococcal coaggregation receptor polysaccharides. AB - The presence of L-rhamnose (Rha) branches in the coaggregation receptor polysaccharides (RPS) of Streptococcus gordonii 38 and Streptococcus oralis J22 was eliminated by replacement of wefB with ermAM in these strains. The expression of this gene in S. oralis 34 did not, however, result in the addition of Rha branches to the linear RPS of this strain, which is identical to that produced by the wefB-deficient mutant of S. gordonii 38. This paradoxical finding was explained by a subtle difference in acceptor specificity of the galactose-1 phosphotransferases encoded by downstream wefC in S. gordonii 38 and wefH in S. oralis 34. These genes were distinguished by the unique ability of WefC to act on the branched acceptor formed by the action of WefB. PMID- 16707705 TI - Transcription of the Bacillus subtilis gerK operon, which encodes a spore germinant receptor, and comparison with that of operons encoding other germinant receptors. AB - The gerA, gerB, and gerK operons, which encode germinant receptors in spores of Bacillus subtilis, were transcribed only in sporulation, and their mRNA levels peaked initially approximately 3 h before the initiation of accumulation of the spore's dipicolinic acid. After a rapid fall, levels of these mRNAs peaked again approximately 5 h later. In one wild-type strain (PS832), gerA mRNA was the most abundant, with levels of gerB and gerK mRNAs approximately 50% of that of gerA mRNA, whereas gerB mRNA was the most abundant in another wild-type strain (PY79). The synthesis of gerK mRNA in sporulation was abolished by loss of the forespore specific RNA polymerase sigma factor, sigma(G), and induction of sigma(G) synthesis in vegetative cells led to synthesis of gerK mRNA. SpoVT, a regulator of sigma(G)-dependent gene expression, repressed gerK expression. The gerK promoter showed sequence similarities to sigma(G)-dependent promoters, and deletion of elements of this putative promoter abolished gerK expression in sporulation. PMID- 16707706 TI - A new integrative conjugative element occurs in Mycoplasma agalactiae as chromosomal and free circular forms. AB - An integrative conjugative element, ICEA, was characterized in Mycoplasma agalactiae strain 5632, in which it occurs as multiple chromosomal copies and as a free circular form. The distribution of ICEA sequences in M. agalactiae strains and their occurrence in Mycoplasma bovis suggest the spreading of the element within or between species. PMID- 16707707 TI - Utilization of the methoxymalonyl-acyl carrier protein biosynthesis locus for cloning the oxazolomycin biosynthetic gene cluster from Streptomyces albus JA3453. AB - Oxazolomycin (OZM), a hybrid peptide-polyketide antibiotic, exhibits potent antitumor and antiviral activities. Using degenerate primers to clone genes encoding methoxymalonyl-acyl carrier protein (ACP) biosynthesis as probes, a 135 kb DNA region from Streptomyces albus JA3453 was cloned and found to cover the entire OZM biosynthetic gene cluster. The involvement of the cloned genes in OZM biosynthesis was confirmed by deletion of a 12-kb DNA fragment containing six genes for methoxymalonyl-ACP biosynthesis from the specific region of the chromosome, as well as deletion of the ozmC gene within this region, to generate OZM-nonproducing mutants. PMID- 16707708 TI - Utilization of the methoxymalonyl-acyl carrier protein biosynthesis locus for cloning of the tautomycin biosynthetic gene cluster from Streptomyces spiroverticillatus. AB - Tautomycin (TTM), a potent protein phosphatase inhibitor, consists of a polyketide chain containing a spiroketal moiety and an acyl chain bearing a dialkylmaleic anhydride structure. PCR using degenerate primers was used to clone genes from Streptomyces spiroverticillatus for formation of the methoxymalonyl acyl carrier protein. This locus was found to contain five genes (ttmC, ttmA, ttmD, ttmB, and ttmE), one of which was used as a probe to clone the 110-kb TTM biosynthetic gene cluster. The involvement of the ttmA gene in TTM biosynthesis was confirmed by gene inactivation and mutation complementation experiments. PMID- 16707709 TI - Xanthosine utilization in Salmonella enterica serovar Typhimurium is recovered by a single aspartate-to-glycine substitution in xanthosine phosphorylase. AB - xapABR from Salmonella enterica was analyzed and compared with the corresponding Escherichia coli genes. xapB and xapR, but not xapA, encode functional proteins. An S. enterica XapA(Asp72Gly) mutant that restores the phosphorolytic activity was selected. The purified mutant enzyme has different kinetic constants than the E. coli enzyme but similar substrate specificity. PMID- 16707710 TI - Gene involved in transcriptional activation of the hrp regulatory gene hrpG in Xanthomonas oryzae pv. oryzae. AB - A novel regulatory gene, trh, which is involved in hrp gene expression, is identified in the plant pathogen Xanthomonas oryzae pv. oryzae. In the trh mutant, expression of HrpG, which is a key regulator for hrp gene expression, is reduced both under the in vitro hrp-inducing condition and in planta. PMID- 16707712 TI - Central resetting of neuromuscular steady states may underlie rhythmical arm movements. AB - Changing the steady-state configuration of the body or its segments may be an important function of central pattern generators for locomotion and other rhythmical movements. Thereby, muscle activation, forces, and movement may emerge following a natural tendency of the neuromuscular system to achieve the current steady-state configuration. To verify that transitions between different steady states occur during rhythmical movements, we asked standing subjects to swing one or both arms synchronously or reciprocally at approximately 0.8 Hz from the shoulder joints. In randomly selected cycles, one arm was transiently arrested by an electromagnetic device. Swinging resumed after some delay and phase resetting. During bilateral swinging, the nonperturbed arm often stopped before resuming swinging at a position that was close to either the extreme forward or the extreme backward arm position observed before the perturbation. Oscillations usually resumed when both arms arrived at similar extreme positions when a synchronous bilateral pattern was initially produced or at the opposite positions if the initial pattern was reciprocal. Results suggest that a central generator controls both arms as a coherent unit by producing transitions between its steady state (equilibrium) positions. By controlling these positions, the system may define the spatial boundaries of movement. At these positions, the system may halt the oscillations, resume them at a new phase (as observed in the present study), or initiate a new motor action. Our findings are relevant to locomotion and suggest that walking may also be generated by transitions between several equilibrium configurations of the body, possibly accomplished by modulation and gating of proprioceptive reflexes. PMID- 16707713 TI - Adaptation in the auditory space map of the barn owl. AB - Auditory neurons in the owl's external nucleus of the inferior colliculus (ICX) integrate information across frequency channels to create a map of auditory space. This study describes a powerful, sound-driven adaptation of unit responsiveness in the ICX and explores the implications of this adaptation for sensory processing. Adaptation in the ICX was analyzed by presenting lightly anesthetized owls with sequential pairs of dichotic noise bursts. Adaptation occurred in response even to weak, threshold-level sounds and remained strong for more than 100 ms after stimulus offset. Stimulation by one range of sound frequencies caused adaptation that generalized across the entire broad range of frequencies to which these units responded. Identical stimuli were used to test adaptation in the lateral shell of the central nucleus of the inferior colliculus (ICCls), which provides input directly to the ICX. Compared with ICX adaptation, adaptation in the ICCls was substantially weaker, shorter lasting, and far more frequency specific, suggesting that part of the adaptation observed in the ICX was attributable to processes resident to the ICX. The sharp tuning of ICX neurons to space, along with their broad tuning to frequency, allows ICX adaptation to preserve a representation of stimulus location, regardless of the frequency content of the sound. The ICX is known to be a site of visually guided auditory map plasticity. ICX adaptation could play a role in this cross-modal plasticity by providing a short-term memory of the representation of auditory localization cues that could be compared with later-arriving, visual-spatial information from bimodal stimuli. PMID- 16707715 TI - Electrophysiological classification of somatostatin-positive interneurons in mouse sensorimotor cortex. AB - Classification of inhibitory interneurons is critical in determining their role in normal information processing and pathophysiological conditions such as epilepsy. Classification schemes have relied on morphological, physiological, biochemical, and molecular criteria; and clear correlations have been demonstrated between firing patterns and cellular markers such as neuropeptides and calcium-binding proteins. This molecular diversity has allowed generation of transgenic mouse strains in which GFP expression is linked to the expression of one of these markers and presumably a single subtype of neuron. In the GIN mouse (EGFP-expressing Inhibitory Neurons), a subpopulation of somatostatin-containing interneurons in the hippocampus and neocortex is labeled with enhanced green fluorescent protein (EGFP). To optimize the use of the GIN mouse, it is critical to know whether the population of somatostatin-EGFP-expressing interneurons is homogeneous. We performed unsupervised cluster analysis on 46 EGFP-expressing interneurons, based on data obtained from whole cell patch-clamp recordings. Cells were classified according to a number of electrophysiological variables related to spontaneous excitatory postsynaptic currents (sEPSCs), firing behavior, and intrinsic membrane properties. EGFP-expressing interneurons were heterogeneous and at least four subgroups could be distinguished. In addition, multiple discriminant analysis was applied to data collected during whole cell recordings to develop an algorithm for predicting the group membership of newly encountered EGFP-expressing interneurons. Our data are consistent with a heterogeneous population of neurons based on electrophysiological properties and indicate that EGFP expression in the GIN mouse is not restricted to a single class of somatostatin-positive interneuron. PMID- 16707714 TI - 5-HT2 receptor activation facilitates a persistent sodium current and repetitive firing in spinal motoneurons of rats with and without chronic spinal cord injury. AB - We examined the modulation of persistent inward currents (PICs) by serotonin (5 HT) in spinal motoneurons of normal and chronic spinal rats. PICs are composed of both a TTX-sensitive persistent sodium current (Na PIC) and a nimodipine sensitive persistent calcium current (Ca PIC), and we focused on quantifying the Na PIC (and its action on the total PIC), which is known to be critical in enabling repetitive firing. Intracellular recordings were made from motoneurons of the whole sacrocaudal spinal cord of normal adult rats after the cord was acutely transected at the S2 spinal level (acute spinal rat condition), removed from the animal, and then maintained in vitro. In vitro motoneuron recordings were likewise made from rats that had a sacral spinal transection 2 mo previously (chronic spinal rats). In motoneurons from acute spinal rats, moderately high doses of 5-HT (> or = 10 microM), or the 5-HT2 receptor agonist DOI (> or = 30 microM), significantly increased the total PIC, hyperpolarized the PIC onset voltage, and hyperpolarized the spike threshold, whereas lower doses had no effect. Both 5-HT and DOI specifically increased the Na PIC portion of the total PIC (tested with nimodipine blocking the Ca PIC). Additionally, 5-HT, but not DOI, depolarized the resting membrane potential (Vm) and increased the input resistance (Rm) in a dose-dependent manner. Therefore 5-HT2 receptor activation facilitated the Na PIC, whereas other 5-HT receptors modulated Vm and Rm. Motoneurons of chronic spinal rats responded to 5-HT and DOI in the same way, but with larger responses and at much lower doses (0.3-1 microM), thus exhibiting a 30-fold supersensitivity to 5-HT. Specifically the Na PIC was supersensitive to 5 HT2 receptor activation with DOI. Also, Rm and Vm were supersensitive to 5-HT. Consistent with the known critical role of the Na PIC in repetitive firing, enhancement of the Na PIC by DOI or 5-HT facilitated the repetitive firing evoked by steady current injection and enabled repetitive firing in a subpopulation of motoneurons of acute spinal rats that were initially unable to produce sustained repetitive firing. We suggest that after spinal transection, residual endogenous spinal sources of 5-HT help facilitate the Na PIC and repetitive firing. With chronic injury, the developed 5-HT supersensitivity more than compensates for lost brain stem 5-HT, so that the Na PIC is large and motoneurons are very excitable, thus contributing to spasticity. PMID- 16707716 TI - Nerve growth factor decreases potassium currents and alters repetitive firing in rat sympathetic neurons. AB - The sympathetic nervous system is an essential regulator of the cardiovascular system and interactions with target tissue regulate sympathetic neuronal properties. The heart produces nerve growth factor (NGF), which promotes sympathetic noradrenergic innervation of cardiac tissue and affects sympathetic synaptic strength. Neurotrophins, including NGF, are important modulators of synaptic plasticity and membrane electrical properties. Here we show that acute application of NGF causes a change in the repetitive firing pattern of cultured sympathetic neurons of the rat superior cervical ganglion. Neurons fire fewer action potentials in NGF, but with increased frequency, demonstrating an NGF dependent change from a tonic to a phasic firing pattern. Additionally, NGF decreases the spike time variance, making spikes more tightly time locked to stimulus onset. NGF causes a decrease in the amplitude of both calcium-dependent and -independent potassium currents, and inhibition of calcium-dependent potassium currents using CdCl(2) reproduces some, but not all, of the firing properties induced by NGF. This study suggests that NGF release from cardiac tissue may act to modulate the repetitive firing properties of sympathetic neurons to tune their output to meet the physiological needs of the organism. PMID- 16707717 TI - Proprioceptive guidance of saccades in eye-hand coordination. AB - The saccade generator updates memorized target representations for saccades during eye and head movements. Here, we tested if proprioceptive feedback from the arm can also update handheld object locations for saccades, and what intrinsic coordinate system(s) is used in this transformation. We measured radial saccades beginning from a central light-emitting diode to 16 target locations arranged peripherally in eight directions and two eccentricities on a horizontal plane in front of subjects. Target locations were either indicated 1) by a visual flash, 2) by the subject actively moving the handheld central target to a peripheral location, 3) by the experimenter passively moving the subject's hand, or 4) through a combination of the above proprioceptive and visual stimuli. Saccade direction was relatively accurate, but subjects showed task-dependent systematic overshoots and variable errors in radial amplitude. Visually guided saccades showed the smallest overshoot, followed by saccades guided by both vision and proprioception, whereas proprioceptively guided saccades showed the largest overshoot. In most tasks, the overall distribution of saccade endpoints was shifted and expanded in a gaze- or head-centered cardinal coordinate system. However, the active proprioception task produced a tilted pattern of errors, apparently weighted toward a limb-centered coordinate system. This suggests the saccade generator receives an efference copy of the arm movement command but fails to compensate for the arm's inertia-related directional anisotropy. Thus the saccade system is able to transform hand-centered somatosensory signals into oculomotor coordinates and combine somatosensory signals with visual inputs, but it seems to have a poorly calibrated internal model of limb properties. PMID- 16707718 TI - GABAA-Receptor blockade reverses the injury-induced sensitization of nociceptor specific (NS) neurons in the spinal dorsal horn of the rat. AB - Single-unit electrical activity was recorded from 80 nociceptor-specific (NS) neurons in the dorsal horn of the lumbar spinal cord of pentobarbital anesthetized rats. Their responses to low- and high-intensity mechanical stimulation of their receptive fields (RFs) were recorded before and after the application of irritant agents [capsaicin (CAP) or mustard oil (MO)] to the RF. Before the applications of the irritants the neurons responded only to high intensity stimuli, but after this procedure 20 of 28 neurons tested were sensitized, i.e., gave increased responses to high-intensity stimuli and showed novel responses to low-intensity mechanical stimulation as well as an Abeta-fiber afferent drive. CAP was more likely to induce sensitization than MO and the majority of sensitized neurons were located in the superficial dorsal horn. No relationship was found between the magnitude of the response to the sensitizing agent and the presence or absence of sensitization. Cumulative doses of two gamma aminobutyric acid type A (GABA(A))-receptor antagonists, picrotoxin and bicuculline, were administered systemically or applied directly over the spinal cord. The GABA(A) antagonists reversed the sensitization of the neurons by reducing the novel low-threshold responses. These results show that NS neurons in the spinal dorsal horn can be sensitized by a sustained afferent discharge in peripheral nociceptors and that this sensitization can be reduced or reversed by low doses of GABA(A)-receptor antagonists. This provides evidence for a mechanism in which an enhanced GABAergic transmission can lead to hyperexcitability and sensitization of NS neurons in the dorsal horn. PMID- 16707720 TI - 5-HT Modulation of identified segmental premotor interneurons in the lamprey spinal cord. AB - Ipsilaterally projecting spinal excitatory interneurons (EINs) generate the hemisegmental rhythmic locomotor activity in lamprey, while the commissural interneurons ensure proper left-right alternation. 5-HT is a potent modulator of the locomotor rhythm and is endogenously released from the spinal cord during fictive locomotion. The effect of 5-HT was investigated for three segmental premotor interneuron types: EINs, commissural excitatory and commissural inhibitory interneurons. All three types of interneurons produced chemical postsynaptic potentials in motoneurons, but only those from EINs had an electrical component. The effect of 5-HT was studied on the slow afterhyperpolarization, involved in spike frequency regulation, and on the segmental synaptic transmission to motoneurons. 5-HT induced a reduction in the slow afterhyperpolarization and a depression of synaptic transmission in all three types of segmental interneurons. Thus 5-HT is a very potent modulator of membrane properties and synaptic transmission of last-order segmental premotor interneurons. Such modulation of locomotor network interneurons can partially account for the observed effects of 5-HT on the swimming pattern in lamprey. PMID- 16707719 TI - Restoration of long-term potentiation in middle-aged hippocampus after induction of brain-derived neurotrophic factor. AB - Restoration of neuronal viability and synaptic plasticity through increased trophic support is widely regarded as a potential therapy for the cognitive declines that characterize aging. Previous studies have shown that in the hippocampal CA1 basal dendritic field deficits in the stabilization of long-term potentiation (LTP) are evident by middle age. The present study tested whether increasing endogenous brain-derived neurotrophic factor (BDNF) could reverse this age-related change. We report here that in middle-aged (8- to 10-mo-old) rats, in vivo treatments with a positive AMPA-type glutamate receptor modulator both increase BDNF protein levels in the cortical telencephalon and restore stabilization of basal dendritic LTP as assessed in acute hippocampal slices 18 h after the last drug treatment. These effects were not attributed to enhanced synaptic transmission or to facilitation of burst responses used to induce LTP. Increasing extracellular levels of BDNF by exogenous application to slices of middle-aged rats was also sufficient to rescue the stabilization of basal dendritic LTP. Finally, otherwise stable LTP in ampakine-treated middle-aged rats can be eliminated by infusion of the extracellular BDNF scavenger TrkB-Fc. Together these results indicate that increases in endogenous BDNF signaling can offset deficits in the postinduction processes that stabilize LTP. PMID- 16707721 TI - Dynamical mechanisms of odor processing in olfactory bulb mitral cells. AB - In the olfactory system, the contribution of dynamical properties such as neuronal oscillations and spike synchronization to the representation of odor stimuli is a matter of substantial debate. While relatively simple computational models have sufficed to guide current research in large-scale network dynamics, less attention has been paid to modeling the membrane dynamics in bulbar neurons that may be equally essential to sensory processing. We here present a reduced, conductance-based compartmental model of olfactory bulb mitral cells that exhibits the complex dynamical properties observed in these neurons. Specifically, model neurons exhibit intrinsic subthreshold oscillations with voltage-dependent frequencies that shape the timing of stimulus-evoked action potentials. These oscillations rely on a persistent sodium conductance, an inactivating potassium conductance, and a calcium-dependent potassium conductance and are reset via inhibitory input such as that delivered by periglomerular cell shunt inhibition. Mitral cells fire bursts, or clusters, of spikes when continuously stimulated. Burst properties depend critically on multiple currents, but a progressive deinactivation of I(A) over the course of a burst is an important regulator of burst termination. Each of these complex properties exhibits appropriate dynamics and pharmacology as determined by electrophysiological studies. Additionally, we propose that a second, inconsistently observed form of infrathreshold bistability in mitral cells may derive from the activation of ATP-activated potassium currents responding to hypoxic conditions. We discuss the integration of these cellular properties in the larger context of olfactory bulb network operations. PMID- 16707722 TI - Motor adaptation to single force pulses: sensitive to direction but insensitive to within-movement pulse placement and magnitude. AB - Although previous experiments have identified that errors in movement induce adaptation, the precise manner in which errors determine subsequent control is poorly understood. Here we used transient pulses of force, distributed pseudo randomly throughout a movement set, to study how the timing of feedback within a movement influenced subsequent predictive control. Human subjects generated a robust adaptive response in postpulse movements that opposed the pulse direction. Regardless of the location or magnitude of the pulse, all pulses yielded similar changes in predictive control. All current supervised and unsupervised theories of motor learning presume that adaptation is proportional to error. Current neural models that broadly encode movement velocity and adapt proportionally to motor error can mimic human insensitivity to pulse location, but cannot mimic human insensitivity to pulse magnitude. We conclude that single trial adaptation to force pulses reveals a categorical strategy that humans adopt to counter the direction, rather than the magnitude, of movement error. PMID- 16707723 TI - Transport of gamma-hydroxybutyrate in rat kidney membrane vesicles: Role of monocarboxylate transporters. AB - Intoxication with gamma-hydroxybutyrate (GHB) is associated with coma, seizure, and death; treatment of overdoses is symptomatic. Previous studies in our laboratory have demonstrated that L-lactate and pyruvate treatment can increase the renal clearance of GHB and increase its elimination in rats, suggesting that GHB may undergo renal reabsorption mediated by monocarboxylic acid transporters (MCTs). The goals of this study were to characterize the renal transport of GHB in rats and to determine the role of MCT in its renal transport. Brush-border membrane (BBM) and basolateral membrane (BLM) vesicles were isolated from rat kidney cortex, and the uptake of L-lactate and GHB was characterized. L-Lactate and GHB undergo both pH- and sodium-dependent transport in BBM vesicles and pH dependent transport in BLM vesicles. A simple Michaelis-Menten equation best described the pH-dependent uptake of GHB in BBM (Km, 8.0 +/- 1.8 mM; Vmax, 838 +/ 45 pmol/mg/s) and in BLM vesicles (Km, 10.5 +/- 2.6 mM; Vmax, 806 +/- 253 pmol/mg/s). mRNA of MCT1 and MCT2 was determined in rat kidney cortex using reverse transcriptase-polymerase chain reaction; using Western blot, the protein expression of MCT1 was present mainly in BLM vesicles, with weak expression in BBM vesicles, whereas that of MCT2 was exclusively in BLM vesicles. Studies with rat MCT1 gene-transfected MDA-MB231 cells demonstrated that GHB was a substrate of MCT1. The data suggest that rat MCT1 may represent an important transporter for GHB in renal tubule cells. This investigation provides evidence for the importance of MCTs in the reabsorption of the monocarboxylic acids l-lactate and GHB in the kidney. PMID- 16707725 TI - Episodic neurologic dysfunction with migraine and reversible imaging findings after radiation. AB - Reported are three adults who developed frequent episodes of a complex neurologic syndrome years after radiation therapy for a brain tumor. MRI and [(18)F]fluorodeoxyglucose PET performed during the episodes demonstrated dramatic gyral thickening with enhancement and intense hypermetabolism in symptomatic regions that resolved on follow-up studies. EEG during episodes showed only slowing. The differential diagnosis and imaging findings are reviewed. PMID- 16707724 TI - CYP4 isoform specificity in the omega-hydroxylation of phytanic acid, a potential route to elimination of the causative agent of Refsum's disease. AB - The saturated C20 isoprenoid phytanic acid is physiologically derived from phytol released in the degradation of chlorophyll. The presence of a C-3 methyl group in this substrate blocks normal beta-oxidation, so phytanic acid degradation primarily occurs by initial peroxisomal alpha-oxidation to shift the register of the methyl group. However, individuals with Refsum's disease are genetically deficient in the required phytanoyl-CoA alpha-hydroxylase and suffer from neurological pathologies caused by the accumulation of phytanic acid. Recent work has shown that phytanic acid can also be catabolized by a pathway initiated by omega-hydroxylation of the hydrocarbon chain, followed by oxidation of the alcohol to the acid and conventional beta-oxidation. However, the enzymes responsible for the omega-hydroxylation of phytanic acid have not been identified. In this study, we have determined the activities of all of the rat and human CYP4A enzymes and two of the rat CYP4F enzymes, with respect to the omega-hydroxylation of phytanic acid. Furthermore, we have shown that the ability to omega-hydroxylate phytanic acid is elevated in microsomes from rats pretreated with clofibrate. The results support a possible role for CYP4 enzyme elevation in the elimination of phytanic acid in Refsum's disease patients. PMID- 16707726 TI - GJA12 mutations in children with recessive hypomyelinating leukoencephalopathy. AB - BACKGROUND: Pelizaeus-Merzbacher-like disease (PMLD) is an inherited hypomyelinating leukoencephalopathy with onset in early infancy. Like Pelizaeus Merzbacher disease (PMD), PMLD is characterized clinically by nystagmus, cerebellar ataxia, and spasticity, due to a permanent lack of myelin deposition in the brain. Mutations in the GJA12 gene, encoding connexin 47 (Cx47), were recently reported in five children with autosomal recessive PMLD. OBJECTIVES: To evaluate the impact of mutations in the GJA12 gene in, and define the clinical and neuroimaging features of, autosomal recessive PMLD. RESULTS: The authors screened for GJA12 mutations in 10 additional PMLD families originating from Italy, Pakistan, and Saudi Arabia. Three novel homozygous GJA12 mutations were identified in 12 mutant cases distributed in 3 of 10 families. The mutations segregated with the disease according to an autosomal recessive trait and included one missense (G236S) and two nonsense (L281fs285X and P131fs144X) changes. CONCLUSIONS: The identification of homozygous mutations predicting the synthesis of aberrant and truncated polypeptides, and their tight segregation with the disease in very large families, clearly demonstrate that the loss of Cx47 function is the cause of the disease. The phenotype of GJA12-related Pelizaeus-Merzbacher-like disease is fairly homogeneous and similar to that of Pelizaeus-Merzbacher disease. However, slower progression of symptoms, greater preservation of cognitive functions, and partial myelination of corticospinal tracts at MRI were distinctive features, which could help in the differential diagnosis. PMID- 16707727 TI - Probable medication-overuse headache: the effect of a 2-month drug-free period. AB - OBJECTIVE: To describe the emerging profile of headache frequency following a 2 month drug-free period in patients with medication overuse. METHODS: All patients treated and discharged from Danish Headache Center in 2002 and 2003 had prospectively filled out a diagnostic headache diary on a daily basis. For patients with probable medication-overuse headache (MOH), the authors determined headache frequencies before and after medication withdrawal from these prospective recordings. RESULTS: Among 1,326 patients, 337 had probable MOH. Two hundred sixteen patients who stayed medication-free for 2 months were eligible. Overall, 45% of the patients improved, 48% had no change, and 7% had more headaches. Median age was 48 years, and male/female ratio 1:2.7. The relative reduction in headache frequency varied considerably with a 67% median reduction in migraine, 0% in tension-type headache (TTH), and 37% in patients with both migraine and TTH (MT). Comparing the diagnostic groups, the relative reduction in frequency differed between migraine and TTH (p < 0.001) and between MT and TTH (p < 0.01). Triptan/ergot overusers improved the most (p < 0.0001). The difference between MT and TTH remained significant when triptan/ergot overuse was controlled (p < 0.05). CONCLUSION: These data demonstrate the benefit of withdrawal in already established medication overuse and support the existence of medication overuse headache as a clinical entity. PMID- 16707728 TI - Long-duration sCJD with PRNP codon 129 methionine homozygosity and cerebral cortical plaques. AB - The authors investigated a 40-year-old woman who presented with ataxia and dementia with little progression for over 40 months. The results of a CSF 14-3-3 protein and EEG study did not reveal major abnormalities. Brain MRI showed increased signal intensity over the occipital cortex in diffusion-weighted imaging. To our knowledge, this is the longest MM-type sporadic Creutzfeldt-Jakob disease case with cortical kuru-type plaques. PMID- 16707729 TI - Denial versus dualism: the blood-brain barrier as an interface of the gut-brain axis. PMID- 16707730 TI - Insulin-like growth factor-I: a treatment for type 2 diabetes revisited. PMID- 16707733 TI - The neural mechanisms underlying the Muller-Lyer illusion and its interaction with visuospatial judgments. AB - Arrows terminating a line can distort the perceived line length. This so-called Muller-Lyer illusion can be used in healthy human subjects to mimic the performance of neglect patients in visuospatial judgments (e.g., in the landmark task). In this study, we investigated the neural mechanisms underlying the Muller Lyer illusion, the landmark task, and their interaction. This was achieved by parametrically manipulating the magnitude of the Muller-Lyer illusion both in a landmark and in a luminance (control) task. As expected, the landmark task activated right posterior parietal cortex and right temporo-occipital cortex. In contrast, the neural processes associated with the strength of the Muller-Lyer illusion were located bilaterally in the lateral occipital cortex as well as the right superior parietal cortex. The data not only converge with but also extend neuropsychological data that indicate maintained line-length illusion in neglect patients. In addition, our results support the size-constancy scaling hypothesis as a putative mechanism underlying line-length illusions. Furthermore, activation that was driven by both the task and the strength of the Muller-Lyer illusion was observed in right intraparietal sulcus, thus arguing in favor of an interaction of illusory information with the top-down processes underlying visuospatial judgments in right parietal cortex. PMID- 16707731 TI - Diabetic hyperphagia--ghrelin in the driver's seat. PMID- 16707734 TI - Frontal-temporal disconnection abolishes object discrimination learning set in macaque monkeys. AB - Two previous studies have shown that frontal-temporal disconnection in monkeys, produced by unilateral ablation of frontal cortex in one hemisphere and of visual inferior temporal cortex in the opposite hemisphere is entirely without effect on visual object-reward association learning in concurrent discrimination tasks. This is a surprising finding in light of the severe impairments that follow frontal-temporal disconnection in many other tests of visual learning and memory, including delayed matching-to-sample and several conditional learning tasks. To explore the limits of this preserved object-reward association learning, we trained monkeys on visual object discrimination learning set (DLS) prior to frontal-temporal disconnection. As a result of training with single object-reward associations, the monkeys acquired a proficient learning set, evidenced by the rapid learning of new single object-reward association problems. This rapid learning was not affected by unilateral ablations of either inferior temporal cortex alone or frontal cortex alone but was severely impaired after final surgery to complete the disconnection. Moreover, each individual monkey now learned single object-reward association problems at the slow rate at which that individual had learned such problems before the formation of learning set. This result shows that frontal-temporal disconnection abolishes visual learning set. PMID- 16707735 TI - Heterosynaptic facilitation of in vivo thalamocortical long-term potentiation in the adult rat visual cortex by acetylcholine. AB - Acetylcholine (ACh) plays a permissive role in developmental plasticity of fibers from the lateral geniculate nucleus (LGN) to the primary visual cortex (V1). These fibers remain plastic and express long-term potentiation (LTP) in adult rodents, but it is not known if ACh modulates this form of plasticity in the mature V1. We show that, in anesthetized rats, theta burst stimulation (TBS) of the LGN using 5 or 40 theta cycles produced moderate (approximately 20%) and stronger (approximately 40%) potentiation, respectively, of field postsynaptic potentials recorded in the ipsilateral V1. Basal forebrain stimulation (100 Hz) 5 min after TBS enhanced LTP induced by both weak (5 theta cycles) and strong (40 theta cycles) induction protocols. Both effects were reduced by systemic administration of the muscarinic receptor antagonist scopolamine. Basal forebrain stimulation did not enhance LTP when applied 30 min after or 5 min prior to TBS, suggesting that ACh affects early LTP induction mechanisms. Application of the cholinergic agonist carbachol in V1 by means of reverse microdialysis mimicked the effect of basal forebrain stimulation. We conclude that heterosynaptic facilitation of V1 plasticity by ACh extends beyond early postnatal maturation periods and acts to convert weak potentiation into pronounced, long-lasting increases in synaptic strength. PMID- 16707736 TI - Group intervention changes brain activity in bilingual language-impaired children. AB - This investigation assessed the effectiveness of a phonological intervention program on the brain functioning of bilingual Finnish 6- to 7-year-old preschool children diagnosed with specific language impairment (SLI). The intervention program was implemented by preschool teachers to small groups of children including children with SLI. A matched group of other bilingual children with SLI received a physical exercise program and served as a control group. Auditory evoked magnetic fields were measured before and after the intervention with an oddball paradigm. The brain activity recordings were followed by a behavioral discrimination test. Our results show that, in children with SLI, the positive intervention effect is reflected in plastic changes in the brain activity of the left and right auditory cortices. PMID- 16707737 TI - A common prefrontal-parietal network for mnemonic and mathematical recoding strategies within working memory. AB - Previous studies have indicated that the lateral prefrontal cortex (LPFC) is closely involved in strategic recoding, even when such processes lessen task demands. For example, 2 studies presented, in the spatial and verbal domains, sequences of stimuli for participants to retain during a short interval and then retrieve. Stimuli were either randomly arranged or structured (forming symmetries and regular shapes for the spatial task and mathematical patterns for the verbal task). Although participants performed the structured tasks better by reorganizing or "chunking" them into more efficient forms, LPFC activity was greater for the structured compared with the random sequences. However, although these results demonstrate that LPFC is involved in strategic recoding, regardless of the type of modality, it remains to be seen whether such a result generalizes to different types of strategic recoding processes. To test this, we presented digit sequence trials that separately emphasized mnemonic or mathematical recoding strategies. While participants were able to gain a performance benefit from either type of recoding strategy, increased LPFC activity was observed for both mathematical and mnemonic recoding trials, compared with either unstructured sequences or control conditions matched for mathematical or mnemonic processes. However, mathematically structured trials activated the LPFC significantly more than mnemonic recoding trials. In addition, lateral posterior parietal cortex was consistently coactivated with LPFC for strategic recoding trials, both in the current experiments and in previous related studies. We conclude that a prefrontal-parietal network is involved in strategic recoding in working memory, regardless of the type of recoding process. PMID- 16707738 TI - Connectivity-Based Parcellation of Broca's Area. AB - It is generally agreed that the cerebral cortex can be segregated into structurally and functionally distinct areas. Anatomical subdivision of Broca's area has been achieved using different microanatomical criteria, such as cytoarchitecture and distribution of neuroreceptors. However, brain function also strongly depends upon anatomical connectivity, which therefore forms a sensible criterion for the functio-anatomical segregation of cortical areas. Diffusion weighted magnetic resonance (MR) imaging offers the opportunity to apply this criterion in the individual living subject. Probabilistic tractographic methods provide excellent means to extract the connectivity signatures from diffusion weighting MR data sets. The correlations among these signatures may then be used by an automatic clustering method to identify cortical regions with mutually distinct and internally coherent connectivity. We made use of this principle to parcellate Broca's area. As it turned out, 3 subregions are discernible that were identified as putative Brodmann area (BA) 44, BA45, and the deep frontal operculum. These results are discussed in the light of previous evidence from other methods in both human and nonhuman primates. We conclude that plausible results can be achieved by the proposed technique, which cannot be obtained by any other method in vivo. For the first time, there is a possibility to investigate the anatomical subdivision of Broca's area noninvasively in the individual living human subject. PMID- 16707739 TI - Dynamics of prefrontal and cingulate activity during a reward-based logical deduction task. AB - We used behavioral and functional magnetic resonance imaging (fMRI) methods to probe the cerebral organization of a simple logical deduction process. Subjects were engaged in a motor trial-and-error learning task, in which they had to infer the identity of an unknown 4-key code. The design of the task allowed subjects to base their inferences not only on the feedback they received but also on the internal deductions that it afforded (autoevaluation). fMRI analysis revealed a large bilateral parietal, prefrontal, cingulate, and striatal network that activated suddenly during search periods and collapsed during ensuing periods of sequence repetition. Fine-grained analyses of the temporal dynamics of this search network indicated that it operates according to near-optimal rules that include 1) computation of the difference between expected and obtained rewards and 2) anticipatory deductions that predate the actual reception of positive reward. In summary, the dynamics of effortful mental deduction can be tracked with fMRI and relate to a distributed network engaging prefrontal cortex and its interconnected cortical and subcortical regions. PMID- 16707740 TI - Selective attention and audiovisual integration: is attending to both modalities a prerequisite for early integration? AB - Interactions between multisensory integration and attention were studied using a combined audiovisual streaming design and a rapid serial visual presentation paradigm. Event-related potentials (ERPs) following audiovisual objects (AV) were compared with the sum of the ERPs following auditory (A) and visual objects (V). Integration processes were expressed as the difference between these AV and (A + V) responses and were studied while attention was directed to one or both modalities or directed elsewhere. Results show that multisensory integration effects depend on the multisensory objects being fully attended--that is, when both the visual and auditory senses were attended. In this condition, a superadditive audiovisual integration effect was observed on the P50 component. When unattended, this effect was reversed; the P50 components of multisensory ERPs were smaller than the unisensory sum. Additionally, we found an enhanced late frontal negativity when subjects attended the visual component of a multisensory object. This effect, bearing a strong resemblance to the auditory processing negativity, appeared to reflect late attention-related processing that had spread to encompass the auditory component of the multisensory object. In conclusion, our results shed new light on how the brain processes multisensory auditory and visual information, including how attention modulates multisensory integration processes. PMID- 16707741 TI - Is there a place for routine imaging for patients in complete remission from aggressive lymphoma? PMID- 16707743 TI - Images in clinical medicine. Burton's line. PMID- 16707744 TI - Can Massachusetts lead the way in health care reform? PMID- 16707745 TI - Health care reform in Massachusetts--a work in progress. PMID- 16707746 TI - Robotic surgery--squeezing into tight places. PMID- 16707747 TI - HER2 and responsiveness of breast cancer to adjuvant chemotherapy. AB - BACKGROUND: Amplification of the human epidermal growth factor receptor type 2 (HER2, also called HER2/neu) gene and overexpression of its product in breast cancer cells may be associated with responsiveness to anthracycline-containing chemotherapy regimens. METHODS: In the randomized, controlled Mammary.5 trial, we studied 639 formalin-fixed paraffin-embedded specimens obtained from 710 premenopausal women with node-positive breast cancer who had received either cyclophosphamide, epirubicin, and fluorouracil (CEF) or cyclophosphamide, methotrexate, and fluorouracil (CMF) as adjuvant chemotherapy. HER2 amplification or overexpression was evaluated with the use of fluorescence in situ hybridization, immunohistochemical analysis, and polymerase-chain-reaction analysis. RESULTS: Amplification of HER2 was associated with a poor prognosis regardless of the type of treatment. In patients whose tumors showed amplification of HER2, CEF was superior to CMF when assessed on the basis of relapse-free survival (hazard ratio, 0.52; 95 percent confidence interval, 0.34 to 0.80; P=0.003) and overall survival (hazard ratio, 0.65; 95 percent confidence interval, 0.42 to 1.02; P=0.06). For women whose tumors lacked amplification of HER2, CEF did not improve relapse-free survival (hazard ratio for relapse, 0.91; 95 percent confidence interval, 0.71 to 1.18; P=0.49) or overall survival (hazard ratio for death, 1.06; 95 percent confidence interval, 0.83 to 1.44; P=0.68). The adjusted hazard ratio for the interaction between treatment and HER2 amplification was 1.96 for relapse-free survival (95 percent confidence interval, 1.15 to 3.36; P=0.01) and 2.04 for overall survival (95 percent confidence interval, 1.14 to 3.65; P=0.02). CONCLUSIONS: Amplification of HER2 in breast cancer cells is associated with clinical responsiveness to anthracycline containing chemotherapy. (cancer.gov number, NCI-V90-0027.). PMID- 16707748 TI - Caffeine therapy for apnea of prematurity. AB - BACKGROUND: Methylxanthines reduce the frequency of apnea of prematurity and the need for mechanical ventilation during the first seven days of therapy. It is uncertain whether methylxanthines have other short- and long-term benefits or risks in infants with very low birth weight. METHODS: We randomly assigned 2006 infants with birth weights of 500 to 1250 g during the first 10 days of life to receive either caffeine or placebo, until drug therapy for apnea of prematurity was no longer needed. We evaluated the short-term outcomes before the first discharge home. RESULTS: Of 963 infants who were assigned to caffeine and who remained alive at a postmenstrual age of 36 weeks, 350 (36 percent) received supplemental oxygen, as did 447 of the 954 infants (47 percent) assigned to placebo (adjusted odds ratio, 0.63; 95 percent confidence interval, 0.52 to 0.76; P<0.001). Positive airway pressure was discontinued one week earlier in the infants assigned to caffeine (median postmenstrual age, 31.0 weeks; interquartile range, 29.4 to 33.0) than in the infants in the placebo group (median postmenstrual age, 32.0 weeks; interquartile range, 30.3 to 34.0; P<0.001). Caffeine reduced weight gain temporarily. The mean difference in weight gain between the group receiving caffeine and the group receiving placebo was greatest after two weeks (mean difference, -23 g; 95 percent confidence interval, -32 to 13; P<0.001). The rates of death, ultrasonographic signs of brain injury, and necrotizing enterocolitis did not differ significantly between the two groups. CONCLUSIONS: Caffeine therapy for apnea of prematurity reduces the rate of bronchopulmonary dysplasia in infants with very low birth weight. (ClinicalTrials.gov number, NCT00182312.). PMID- 16707749 TI - Volume progression in polycystic kidney disease. AB - BACKGROUND: Autosomal dominant polycystic kidney disease (ADPKD) is characterized by progressive enlargement of cyst-filled kidneys. METHODS: In a three-year study, we measured the rates of change in total kidney volume, total cyst volume, and iothalamate clearance in patients with ADPKD. Of a total of 241 patients, in 232 patients without azotemia who were 15 to 46 years old at baseline we used magnetic-resonance imaging to correlate the total kidney volume and total cyst volume with iothalamate clearance. Statistical methods included analysis of variance, Pearson correlation, and multivariate regression analysis. RESULTS: Total kidney volume and total cyst volume increased exponentially, a result consistent with an expansion process dependent on growth. The mean (+/-SD) total kidney volume was 1060+/-642 ml at baseline and increased by a mean of 204+/-246 ml (5.27+/-3.92 percent per year, P<0.001) over a three-year period among 214 patients. Total cyst volume increased by 218+/-263 ml (P<0.001) during the same period among 210 patients. The baseline total kidney volume predicted the subsequent rate of increase in volume, independently of age. A baseline total kidney volume above 1500 ml in 51 patients was associated with a declining glomerular filtration rate (by 4.33+/-8.07 ml per minute per year, P<0.001). Total kidney volume increased more in 135 patients with PKD1 mutations (by 245+/ 268 ml) than in 28 patients with PKD2 mutations (by 136+/-100 ml, P=0.03). CONCLUSIONS: Kidney enlargement resulting from the expansion of cysts in patients with ADPKD is continuous and quantifiable and is associated with the decline of renal function. Higher rates of kidney enlargement are associated with a more rapid decrease in renal function. PMID- 16707750 TI - Language ability after early detection of permanent childhood hearing impairment. AB - BACKGROUND: Children with bilateral permanent hearing impairment often have impaired language and speech abilities. However, the effects of universal newborn screening for permanent bilateral childhood hearing impairment and the effects of confirmation of hearing impairment by nine months of age on subsequent verbal abilities are uncertain. METHODS: We studied 120 children with bilateral permanent hearing impairment identified from a large birth cohort in southern England, at a mean of 7.9 years of age. Of the 120 children, 61 were born during periods with universal newborn screening and 57 had hearing impairment that was confirmed by nine months of age. The primary outcomes were language as compared with nonverbal ability and speech expressed as z scores (the number of standard deviations by which the score differed from the mean score among 63 age-matched children with normal hearing), adjusted for the severity of the hearing impairment and for maternal education. RESULTS: Confirmation of hearing impairment by nine months of age was associated with higher adjusted mean z scores for language as compared with nonverbal ability (adjusted mean difference for receptive language, 0.82; 95 percent confidence interval, 0.31 to 1.33; and adjusted mean difference for expressive language, 0.70; 95 percent confidence interval, 0.13 to 1.26). Birth during periods with universal newborn screening was also associated with higher adjusted z scores for receptive language as compared with nonverbal ability (adjusted mean difference, 0.60; 95 percent confidence interval, 0.07 to 1.13), although the z scores for expressive language as compared with nonverbal ability were not significantly higher. Speech scores did not differ significantly between those who were exposed to newborn screening or early confirmation and those who were not. CONCLUSIONS: Early detection of childhood hearing impairment was associated with higher scores for language but not for speech in midchildhood. PMID- 16707751 TI - Clinical practice. Acute pancreatitis. PMID- 16707752 TI - Newborn hearing screening--a silent revolution. PMID- 16707753 TI - Images in clinical medicine. Sympathetic-nerve damage in brachial-plexus palsy. PMID- 16707754 TI - Case records of the Massachusetts General Hospital. Case 15-2006. A 46-year-old woman with sudden onset of abdominal distention. PMID- 16707755 TI - Anthracyclines and the tailoring of treatment for early breast cancer. PMID- 16707756 TI - Caffeine for apnea of prematurity. PMID- 16707757 TI - Imaging progression in polycystic kidney disease. PMID- 16707758 TI - Glucosamine and chondroitin sulfate for knee osteoarthritis. PMID- 16707759 TI - Trastuzumab treatment in breast cancer. PMID- 16707760 TI - Cetuximab plus radiotherapy for head and neck cancer. PMID- 16707761 TI - Selective serotonin-reuptake inhibitors and persistent pulmonary hypertension of the newborn. PMID- 16707762 TI - Mortality after the hospitalization of a spouse. PMID- 16707763 TI - Drug-related hepatotoxicity. PMID- 16707764 TI - Gefitinib-sensitizing mutations in esophageal carcinoma. PMID- 16707765 TI - Magnetic resonance imaging for the detection of myocardial fibrosis in scleroderma. PMID- 16707767 TI - Regional expression of prostaglandin E2 and F2alpha receptors in human myometrium, amnion, and choriodecidua with advancing gestation and labor. AB - The change from uterine quiescence to enhanced contractile activity may be due to the differential expression of prostaglandin receptors within the myometrium and fetal membranes, in a temporal and topographically distinct manner. To address this question, we determined the localization and expression of the PGE2 receptor subtypes (PTGER1-4) and the PGF2alpha receptor (PTGFR) in paired upper and lower segment myometrium, amnion, and choriodecidual samples throughout human pregnancy, with and without labor. All receptor subtypes were found throughout the muscle layers in both the upper and lower uterine segments, colocalizing with alpha smooth muscle actin. A change in intracellular localization was observed at term labor, where PTGER1 and PTGER4 were predominately associated with the nucleus. Minimal changes in the expression of the PGE2 and PGF2alpha receptor subtypes were observed with gestational age, labor, or between the upper and lower myometrial segments. Receptor expression in maternal and fetal tissues differed between the receptor subtypes; PTGER1 and PTGER4 were predominately expressed in the fetal membranes, PTGER2 was greatest in the myometrium, whereas PTGER3 and PTGFR were similarly expressed in the myometrium and fetal membranes. Myometrial activation through the prostaglandin receptors is perhaps more subtle and may be mediated by a balance between one or several of the prostaglandin receptor subtypes together with other known contraction associated proteins. Lack of coordination in receptor expression between the myometrium and fetal membranes may indicate different regulatory mechanisms between these tissues, or it may suggest a function for these receptors in the amnion and choriodecidua that is independent of that seen in the myometrium. PMID- 16707766 TI - Progesterone regulation of preimplantation conceptus growth and galectin 15 (LGALS15) in the ovine uterus. AB - Peri-implantation conceptus (embryo/fetus and associated extraembryonic membranes) growth and development are primarily regulated by secretions from the uterus. This study investigated the effects of progesterone on preimplantation conceptus development and endometrial galectin 15 (LGALS15). Ewes received daily injections of either corn oil (CO) vehicle or 25 mg progesterone (P4) from 36 h postmating to hysterectomy. Treatment with P4 increased blastocyst diameter by 220% on Day 9 and advanced time of elongation of blastocysts to a filamentous conceptus on Day 12. Effects of P4 treatment on blastocyst development were blocked by administration of RU486, a progesterone receptor antagonist. Consistent with early elongation of blastocysts, interferon tau (IFNT) protein was about 50-fold greater in uterine flushes from Day 12 in ewes receiving P4 compared with those receiving CO. Expression of cathepsin L (CTSL) and radical S adenosyl methionine domain containing 2 (RSAD2), both IFNT-stimulated genes, was increased in endometria of Day 12 P4-treated ewes. LGALS15 mRNA, expressed only in the endometrial luminal epithelium and superficial glands, was detected between Days 9 and 12 and was more abundant in ewes receiving P4 than in those receiving CO on both Days 9 and 12. RU486 treatment ablated P4 induction of LGALS15 mRNA in the endometrial epithelia. LGALS15 protein in uterine flushings was not different on Day 9 but tended to be greater in P4-treated ewes than in those receiving CO on Day 12. The advanced development of blastocysts in P4 treated ewes is hypothesized to involve early induction of specific genes in the endometrial epithelia, such as LGALS15, and undoubtedly components of uterine histotroph. PMID- 16707768 TI - ECC-1 cells: a well-differentiated steroid-responsive endometrial cell line with characteristics of luminal epithelium. AB - Endometrial cancer cell lines have provided a valuable model to study endometrial epithelial cells in vitro. Since the first development of HEC1B over 35 yr ago, many different cell lines have been isolated and described. One valuable cell line that maintains hormone responsiveness and unique stability over time is the ECC-1 cell line, developed originally by the late P.G. Satyaswaroop. In this study, we investigated some of the properties of these cells and present their salient characteristics. Like Ishikawa cells, ECC-1 cells maintain both estrogen receptors (ESR1 [ER alpha] and ESR2 [ER beta]), progesterone receptors (PR A and B; PGRs), and androgen receptors (ARs), along with the p160 steroid receptor coactivators NCOA1 (formerly SRC1), NCOA2 (formerly TIF2), and NCOA3 (formerly AIB1). The karyotype of these cells is abnormal, with multiple structural rearrangements in all cells analyzed. Unlike Ishikawa cells that express glandular epithelial antigens, ECC-1 cells maintain a luminal phenotype, with expression of KRT13 (cytokeratin 13) and KRT18 (cytokeratin 18). Apparent differences in the regulation of ESR2 also were evident in ECC-1 cells compared to Ishikawa cells. Like other endometrial cell lines, ECC-1 cells express the steroid receptor coactivators and exhibit epidermal growth factor-stimulated expression of known luminal proteins thought to be involved in implantation, including the hyaluronate receptor CD44 and SPP1 (formerly osteopontin) and CD55 (decay-accelerating factor). These characteristics appear to be stable and persistent over multiple cell passages, making this well-differentiated cell line an excellent choice to study endocrine and paracrine regulation of endometrial epithelium in vitro. PMID- 16707769 TI - Testosterone stimulates growth and secretory activity of the female prostate in the adult gerbil (Meriones unguiculatus). AB - The prostate of the female gerbil (Meriones unguiculatus) is similar to the human female prostate (Skene gland) and, despite its reduced size, it is functional and shows secretory activity. However, virtually nothing is known about its physiological regulation. This study was thus undertaken to evaluate the behavior of the gerbil female prostate in a hyperandrogenic condition. Adult females received subcutaneous injections of testosterone cypionate (1 mg/kg body weight every 48 h) up to 21 days. Circulating levels of testosterone and estradiol were monitored, and the prostate and ovaries subjected to structural and immunocytochemical analyses. The treatment resulted in sustained high levels of circulating testosterone, and caused a transient increase in estradiol. There was an increase in epithelial cell proliferation accompanied by significant reorganization of the epithelium and an apparent reduction in secretory activity, followed by a progressive increase in luminal volume density and accumulation of secretory products. Immunocytochemistry identified the expression of androgen receptor and a prostate-specific antigen (PSA)-related antigen in prostatic epithelial cells. A circulating PSA-related antigen was also found, and its concentration showed strong negative correlation with circulating estrogen. Epithelial dysplasia was detected in the prostate of treated females. Analysis of the ovaries showed the occurrence of a polycystic condition and stromal cell hyperplasia. The results indicate that testosterone has a stimulatory effect on the female prostate, inducing epithelial cell proliferation, differentiation, secretory activity, and dysplasia. The results also suggest that prostatic growth and activity, polycystic ovaries, and ovarian stromal cell hyperplasia are related to a hyperandrogenic condition in females. PMID- 16707770 TI - Reactivity of human placental chorionic plate vessels from pregnancies complicated by intrauterine growth restriction (IUGR). AB - A successful pregnancy is dependent on liberal placental perfusion via the maternal and fetal circulations. Doppler waveform analyses of umbilical arteries suggest increased resistance to flow in the fetoplacental circulation of pregnancies complicated by intrauterine growth restriction (IUGR). Neither the site nor the mediators responsible for this altered vascular reactivity are known, to date. In placentas in normal pregnancy, reduced oxygenation promotes contraction of the in vitro-perfused placental cotyledon and modulates agonist induced contraction of chorionic plate arteries and veins. Placental oxygenation has also been suggested to be reduced in IUGR. We tested the hypothesis that oxygen tension could directly modify placental chorionic plate vessel vasoreactivity in IUGR. Small arteries and veins from the chorionic plate were dissected from biopsies from placentas of pregnancies complicated by IUGR and were studied using parallel wire myography. Vasoconstriction at 20%, 7%, and 2% oxygen was assessed utilizing the thromboxane mimetic U46619. Experiments were also performed in the presence of 4-aminopyridine (4AP), a blocker of voltage gated potassium channels. Increased oxygenation reduced venous vasoconstriction but did not modify arterial vasoconstriction. 4AP increased basal tone in arteries and veins. We suggest that venoconstriction in response to hypoxia may provide a mechanism for increased fetoplacental vascular resistance associated with IUGR. PMID- 16707771 TI - Regulation of the stimulant actions of neurokinin a and human hemokinin-1 on the human uterus: a comparison with histamine. AB - Regulation of the contractile effects of tachykinins and histamine on the human uterus was investigated with biopsy sections of the outer myometrial layer. The effects of neurokinin A (NKA) and human hemokinin-1 (hHK-1) in tissues from pregnant but not from nonpregnant women were enhanced by the inhibition of neprilysin. The effects of NKA and eledoisin were blocked by the NK2 receptor antagonist SR 48968 but not by the NK1 receptor antagonist SR 140333 in tissues from both groups of women. Human HK-1 acted as a partial agonist blocked by SR 48968 and, to a lesser extent, by SR 140333; endokinin D was inactive. In tissues from pregnant women, responses to high potassium-containing Krebs solution were 2 3-fold higher than those from nonpregnant women. Mepyramine-sensitive maximal responses to histamine were similarly enhanced. The absolute maximum responses to NKA and its stable NK2 receptor-selective analogue, [Lys5MeLeu9Nle10]NKA(4-10), were increased in pregnancy, but their efficacies relative to potassium responses were decreased. Tachykinin potencies were lower in tissues from pregnant women than in those from nonpregnant women. These data 1) show for the first time that hHK-1 is a uterine stimulant in the human, 2) confirm that the NK2 receptor is predominant in mediating tachykinin actions on the human myometrium, and 3) indicate that mammalian tachykinin effects are tightly regulated during pregnancy in a manner that would negate an inappropriate uterotonic effect. The potencies of these peptides in tissues from nonpregnant women undergoing hysterectomy are consistent with their possible role in menstrual and menopausal disorders. PMID- 16707772 TI - Photoperiodic regulation of compensatory testicular hypertrophy in hamsters. AB - In mammals, removal of one testis results in compensatory testicular hypertrophy (CTH) of the remaining gonad. Although CTH is ubiquitous among juveniles of many species, laboratory rats, laboratory mice, and humans unilaterally castrated in adulthood fail to display CTH. We documented CTH in pre- and postpubertally hemi castrated Syrian and Siberian hamsters and tested whether day length affects CTH in juvenile and adult Siberian hamsters. Robust CTH was evident in long-day hemi castrates of both species and was preceded by increased serum FSH concentrations in juvenile Siberian hamsters. In sharp contrast, CTH was undetectable in short day hemi-castrated Siberian hamsters for several months and only made its appearance with the development of neuroendocrine refractoriness to short day lengths; serum FSH concentrations of juveniles also did not increase above sham castrate values until the onset of refractoriness. Long-day hemi-castrated Siberian hamsters with hypertrophied testes underwent complete gonadal regression after transfer to short days, albeit at a reduced rate for the first 3 weeks of treatment. Blood testosterone concentrations of adult hamsters did not differ between long-day hemicastrates and sham-castrates 9-12 weeks after surgery. We conclude that CTH is suppressed by short day lengths in Siberian hamsters at all ages and stages of reproductive development; in short day lengths, but not long day lengths, the remaining testis produces sufficient negative feedback inhibition to restrain FSH hypersecretion and prevent CTH. PMID- 16707773 TI - Identification and characterization of novel and unknown mouse epididymis specific genes by complementary DNA microarray technology. AB - To examine epididymal function, we attempted to identify highly expressed genes in mouse epididymis using a cDNA microarray containing PCR products amplified from a mouse epididymal cDNA library. We isolated one novel and four known genes lymphocyte cytosolic protein 1 (Lcp1), complement subcomponents C1r/C1s, Uegf protein, and bone morphogenetic protein and zona pellucida-like domains 1 (Cuzd1), transmembrane epididymal protein 1 (Teddm1), and whey acidic protein 4 disulfide core domain 16 (Wfdc16)-with unknown functions in the epididymis. The novel gene, designated Serpina1f (serine peptidase inhibitor [SERPIN], clade A, member 1f), harbors an open reading frame of 1 233 bp encoding a putative protein of 411 amino acids, including a SERPIN domain. These five genes were predominantly expressed in the epididymis as compared to other organs. In situ hybridization analysis revealed their epididymal region-specific expression patterns. Real-time RT-PCR analysis revealed a significant increase in mRNA expression of these genes around puberty. Castration decreased their expression, except forLcp1. Testosterone (T) restored these reduced expressions, except forTeddm1; however, this restoration was not observed with 17 beta-estradiol (E2). Administration of T and E2 combination recovered the Serpina1f mRNA concentration; this recovery was also observed with T alone. However, the recovery of Cuzd1and Wfdc16mRNA concentrations was inadequate. Neonatal diethylstilbestrol treatment suppressed the Cuzd1, Wfdc16, and Serpina1f mRNA expression in the epididymis of 8-week-old mice; this was not observed with E2. These results suggest that our microarray system can provide a novel insight into the epididymal function on a molecular basis, and the five genes might play important roles in the epididymis. PMID- 16707774 TI - Editorial--modeling negative symptoms: what's missing? PMID- 16707775 TI - At issue: is natural selection rendering schizophrenia less severe? PMID- 16707776 TI - Why having a mental illness is not like having diabetes. PMID- 16707779 TI - Perceptions of physical and social environment variables and self-efficacy as correlates of self-reported physical activity among adolescent girls. AB - OBJECTIVE: This cross-sectional study examined the direct and indirect effects of perceived equipment accessibility, neighborhood safety, and social support on self-reported physical activity among older adolescent girls. METHODS: Adolescent girls (n = 1,655) who were in the 12th grade completed a battery of questionnaires that included self-report measures of the perceived physical environment, social support, barriers self-efficacy, and physical activity. RESULTS: Perceived neighborhood safety did not exhibit direct or indirect effects on self-reported physical activity. Perceived equipment accessibility exhibited an indirect effect on self-reported physical activity that was accounted for by barriers self-efficacy. Perceived social support exhibited direct and indirect effects on self-reported physical activity; the indirect effect was accounted for by barriers self-efficacy. The relationships were independent of parental education and body mass index (BMI). CONCLUSIONS: Perceived physical environmental factors indirectly influenced self-reported physical activity, and perceived social environmental factors both directly and indirectly influenced self-reported physical activity in this sample of older adolescent girls. PMID- 16707777 TI - Neuropsychological functioning in adolescents and young adults at genetic risk for schizophrenia and affective psychoses: results from the Harvard and Hillside Adolescent High Risk Studies. AB - Siblings and offspring of persons with schizophrenia carry elevated genetic risk for the illness and manifest attentional and memory impairments. Because less is known about other neuropsychological functions and their specificity in adolescents, we conducted a genetic high-risk (HR) study of schizophrenia (HR SCZ) and affective psychosis (HR-AFF). Participants (ages 12-25) were from the Harvard Adolescent High-Risk and Hillside Family studies, including 73 HR-SCZ, 18 HR-AFF, and 84 community controls (CCs) recruited in metropolitan Boston and New York. Groups were compared on overall neurocognitive functioning, 6 domains, and 13 test scores, controlling for age, parental education, and correlated data within families. The HR-SCZ group was significantly impaired overall, while the HR-AFF group demonstrated a trend toward overall impairment. HR-SCZ subjects showed significantly lower Verbal Ability (d = .73) and Executive Functioning/Working Memory (d = .47) than CCs. HR-AFF subjects showed reduced Verbal Ability (d = .64) compared to CCs. Excluding 12 CCs with a parental history of depression (without psychosis) led to larger differences between HR and CC groups across domains. Moreover, HR-SCZ and CC group differences in Verbal Memory (d = .39) and Visual-Spatial (d = .34) became statistically significant. There were no significant differences between HR-SCZ and HR-AFF groups. Data support a modest neuropsychological deficit in persons at genetic HR for psychosis, with a broader range of deficits in HR-SCZ. Future work should assess the relationship of neurocognition to adaptive functioning and possible onset of psychosis in HR samples. Ascertainment criteria for controls may markedly influence results and interpretation of group differences. PMID- 16707780 TI - Combined application of behavior genetics and microarray analysis to identify regional expression themes and gene-behavior associations. AB - In this report we link candidate genes to complex behavioral phenotypes by using a behavior genetics approach. Gene expression signatures were generated for the prefrontal cortex, ventral striatum, temporal lobe, periaqueductal gray, and cerebellum in eight inbred strains from priority group A of the Mouse Phenome Project. Bioinformatic analysis of regionally enriched genes that were conserved across all strains revealed both functional and structural specialization of particular brain regions. For example, genes encoding proteins with demonstrated anti-apoptotic function were over-represented in the cerebellum, whereas genes coding for proteins associated with learning and memory were enriched in the ventral striatum, as defined by the Expression Analysis Systematic Explorer (EASE) application. Association of regional gene expression with behavioral phenotypes was exploited to identify candidate behavioral genes. Phenotypes that were investigated included anxiety, drug-naive and ethanol-induced distance traveled across a grid floor, and seizure susceptibility. Several genes within the glutamatergic signaling pathway (i.e., NMDA/glutamate receptor subunit 2C, calmodulin, solute carrier family 1 member 2, and glutamine synthetase) were identified in a phenotype-dependent and region-specific manner. In addition to supporting evidence in the literature, many of the genes that were identified could be mapped in silico to surrogate behavior-related quantitative trait loci. The approaches and data set described herein serve as a valuable resource to investigate the genetic underpinning of complex behaviors. PMID- 16707778 TI - Defining and assessing adherence to oral antipsychotics: a review of the literature. AB - The definition and assessment of adherence vary considerably across studies. Increasing consensus regarding these issues is necessary to improve our understanding of adherence and the development of more effective treatments. We review the adherence literature over the past 3 decades to explore the definitions and assessment of adherence to oral antipsychotics in schizophrenia patients. A total of 161 articles were identified through MEDLINE and PsycINFO searches. The most common method used to assess adherence was the report of the patient. Subjective and indirect methods including self-report, provider report, significant other report, and chart review were the only methods used to assess adherence in over 77% (124/161) of studies reviewed. Direct or objective measures including pill count, blood or urine analysis, electronic monitoring, and electronic refill records were used in less than 23% (37/161) of studies. Even in studies utilizing the same methodology to assess adherence, definitions of an adherent subject varied broadly from agreeing to take any medication to taking at least 90% of medication as prescribed. We make suggestions for consensus development, including the use of recommended terminology for different subject samples, the increased use of objective or direct measures, and the inclusion in all studies of an estimate of the percentage of medication taken as prescribed in an effort to increase comparability among studies. The suggestions are designed to advance the field with respect to both understanding predictors of adherence and developing interventions to improve adherence to oral antipsychotic medications. PMID- 16707781 TI - Small, nonpeptide p75NTR ligands induce survival signaling and inhibit proNGF induced death. AB - Studies showing that neurotrophin binding to p75NTR can promote cell survival in the absence of Trk (tropomyosin-related kinase) receptors, together with recent structural data indicating that NGF may bind to p75NTR in a monovalent manner, raise the possibility that small molecule p75NTR ligands that positively regulate survival might be found. A pharmacophore designed to capture selected structural and physical chemical features of a neurotrophin domain known to interact with p75NTR was applied to in silico screening of small molecule libraries. Small, nonpeptide, monomeric compounds were identified that interact with p75NTR. In cells showing trophic responses to neurotrophins, the compounds promoted survival signaling through p75NTR-dependent mechanisms. In cells susceptible to proneurotrophin-induced death, compounds did not induce apoptosis but inhibited proneurotrophin-mediated death. These studies identify a unique range of p75NTR behaviors that can result from isolated receptor liganding and establish several novel therapeutic leads. PMID- 16707782 TI - Spatiotemporal tuning of rapid interactions between visual-motion analysis and reaching movement. AB - In addition to the goal-directed preplanned control, which strongly governs reaching movements, another type of control mechanism is suggested by recent findings that arm movements are rapidly entrained by surrounding visual motion. It remains, however, controversial whether this rapid manual response is generated in a goal-oriented manner similarly to preplanned control or is reflexively and directly induced by visual motion. To investigate the sensorimotor process underlying rapid manual responses induced by large-field visual motion, we examined the effects of contrast and spatiotemporal frequency of the visual-motion stimulus. The manual response amplitude increased steeply with image contrast up to 10% and leveled off thereafter. Regardless of the spatial frequency, the response amplitude increased almost proportionally to the logarithm of stimulus speed until the temporal frequency reached 15-20 Hz and then fell off. The maximum response was obtained at the lowest spatial frequency we examined (0.05 cycles/degrees). These stimulus specificities are surprisingly similar to those of the reflexive ocular-following response induced by visual motion, although there is no direct motor entrainment from the ocular to manual responses. In addition, the spatiotemporal tuning is clearly different from that of perceptual effects caused by visual motion. These comparisons suggest that the rapid manual response is generated by a reflexive sensorimotor mechanism. This mechanism shares a distinctive visual-motion processing stage with the reflexive control for other motor systems yet is distinct from visual-motion perception. PMID- 16707783 TI - Ischemic insults direct glutamate receptor subunit 2-lacking AMPA receptors to synaptic sites. AB - Regulated AMPA receptor (AMPAR) trafficking at excitatory synapses is a mechanism critical to activity-dependent alterations in synaptic efficacy. The role of regulated AMPAR trafficking in insult-induced synaptic remodeling and/or cell death is, however, as yet unclear. Here we show that brief oxygen-glucose deprivation (OGD), an in vitro model of brain ischemia, promotes redistribution of AMPARs at synapses of hippocampal neurons, leading to a switch in AMPAR subunit composition. Ischemic insults promote internalization of glutamate receptor subunit 2 (GluR2)-containing AMPARs from synaptic sites via clathrin dependent endocytosis and facilitate delivery of GluR2-lacking AMPARs to synaptic sites via soluble N-ethylmaleimide-sensitive factor attachment protein receptor dependent exocytosis, evident at early times after insult. The OGD-induced switch in receptor subunit composition requires PKC activation, dissociation of GluR2 from AMPA receptor-binding protein, and association with protein interacting with C kinase-1. We further show that AMPARs at synapses of insulted neurons exhibit functional properties of GluR2-lacking AMPARs. AMPAR-mediated miniature EPSCs exhibit increased amplitudes and enhanced sensitivity to subunit-specific blockers of GluR2-lacking AMPARs, evident at 24 h after ischemia. The OGD-induced alterations in synaptic AMPA currents require clathrin-mediated receptor endocytosis and PKC activation. Thus, ischemic insults promote targeting of GluR2 lacking AMPARs to synapses of hippocampal neurons, mechanisms that may be relevant to ischemia-induced synaptic remodeling and/or neuronal death. PMID- 16707784 TI - Activity of Renshaw cells during locomotor-like rhythmic activity in the isolated spinal cord of neonatal mice. AB - In the present study, we examine the activity patterns of and synaptic inputs to Renshaw cells (RCs) during fictive locomotion in the newborn mouse using visually guided recordings from GABAergic cells expressing glutamic acid decarboxylase 67 green fluorescent protein (GFP). Among the GFP-positive neurons in the lumbar ventral horn, RCs were uniquely identified as receiving ventral root-evoked short latency EPSPs that were markedly reduced in amplitude by nicotinic receptor blockers mecamylamine or tubocurarine. During locomotor-like rhythmic activity evoked by bath application of 5-HT and NMDA, 50% of the recorded RCs fired in phase with the ipsilateral L2 flexor-related rhythm, whereas the rest fired in the extensor phase. Each population of RCs fired throughout the corresponding locomotor phase. All RCs received both excitatory and inhibitory synaptic inputs during the locomotor-like rhythmic activity. Blocking nicotinic receptors with mecamylamine markedly reduced the rhythmic excitatory drive, indicating that these rhythmic inputs originate mainly from motor neurons (MNs). Inhibitory synaptic inputs persisted in the presence of the nicotinic blocker. Part of this inhibitory drive and remaining excitatory drive could be from commissural interneurons because the present study also shows that RCs receive direct crossed inhibitory and excitatory synaptic inputs. However, rhythmic synaptic inputs in RCs were also observed in hemicord preparations in the presence of mecamylamine. These results show that, during locomotor activity, RC firing properties are modulated not only by MNs but also by the ipsilateral and contralateral locomotor networks. PMID- 16707786 TI - Deglycosylated anti-amyloid-beta antibodies eliminate cognitive deficits and reduce parenchymal amyloid with minimal vascular consequences in aged amyloid precursor protein transgenic mice. AB - Systemic administration of anti-amyloid-beta (Abeta) antibodies results in reduced parenchymal amyloid but increased vascular amyloid and microhemorrhage in amyloid precursor protein (APP) transgenic mice. Here, we evaluate the effects of reducing effector interactions of the antibody via deglycosylation. Mice aged 20 months were treated weekly for 4 months and tested behaviorally before they were killed. APP transgenic mice receiving either anti-Abeta (2H6) or deglycosylated anti-Abeta (de-2H6) showed significant improvement in radial arm water maze performance compared with mice receiving a control antibody. Both groups receiving anti-Abeta antibodies showed significant reductions in total Abeta immunochemistry and Congo red. Significantly fewer vascular amyloid deposits and microhemorrhages were observed in mice administered the de-2H6 antibody compared with those receiving unmodified 2H6 antibody. Deglycosylated anti-Abeta antibodies may be preferable to unmodified IgG because they retain the cognition enhancing and amyloid-reducing properties of anti-Abeta immunotherapy, while greatly attenuating the increased vascular amyloid deposition and microhemorrhage observed with unmodified IgG. PMID- 16707785 TI - ASIC1a-specific modulation of acid-sensing ion channels in mouse cortical neurons by redox reagents. AB - Acid-sensing ion channel (ASIC)-1a, the major ASIC subunit with Ca2+ permeability, is highly expressed in the neurons of CNS. Activation of these channels with resultant intracellular Ca2+ accumulation plays a critical role in normal synaptic plasticity, learning/memory, and in acidosis-mediated glutamate receptor-independent neuronal injury. Here we demonstrate that the activities of ASICs in CNS neurons are tightly regulated by the redox state of the channels and that the modulation is ASIC1a subunit dependent. In cultured mouse cortical neurons, application of the reducing agents dramatically potentiated, whereas the oxidizing agents inhibited the ASIC currents. However, in neurons from the ASIC1 knock-out mice, neither oxidizing agents nor reducing reagents had any effect on the acid-activated current. In Chinese Hamster Ovary cells, redox-modifying agents only affected the current mediated by homomeric ASIC1a, but not homomeric ASIC1b, ASIC2a, or ASIC3. In current-clamp recordings and Ca(2+)-imaging experiments, the reducing agents increased but the oxidizing agents decreased acid-induced membrane depolarization and the intracellular Ca2+ accumulation. Site-directed mutagenesis studies identified involvement of cysteine 61 and lysine 133, located in the extracellular domain of the ASIC1a subunit, in the modulation of ASICs by oxidizing and reducing agents, respectively. Our results suggest that redox state of the ASIC1a subunit is an important factor in determining the overall physiological function and the pathological role of ASICs in the CNS. PMID- 16707787 TI - Genes associated with adult axon regeneration promoted by olfactory ensheathing cells: a new role for matrix metalloproteinase 2. AB - The molecular mechanisms used by olfactory ensheathing cells (OECs) to promote repair in the damaged adult mammalian CNS remain unknown. Thus, we used microarrays to analyze three OEC populations with different capacities to promote axonal regeneration in cultured rat retinal neurons. Gene expression in "long term cultured OECs" that do not stimulate adult axonal outgrowth was compared with that of "primary olfactory ensheathing cells" and the immortalized OEC cell line TEG3. In this way, we identified a number of candidate genes that might play a role in promoting adult axonal regeneration. Among these genes, it was striking that both the matrix metalloproteinase 2 (MMP2) and an inhibitor of this protease were represented. The disruption of MMP2 activity in TEG3 cells impaired their capacity to trigger axon regeneration in cultured adult retinal neurons. Furthermore, the MMP2 protein was detected in grafts of OECs that elicited robust axonal regeneration in the injured spinal cord of adult rats in vivo. These data suggest that MMP2 does indeed participate in adult axonal regeneration induced by OECs. PMID- 16707788 TI - Role of dopamine in the primate caudate nucleus in reward modulation of saccades. AB - Expected reward impacts behavior and neuronal activity in brain areas involved in sensorimotor processes. However, where and how reward signals affect sensorimotor signals is unclear. Here, we show evidence that reward-dependent modulation of behavior depends on normal dopamine transmission in the striatum. Monkeys performed a visually guided saccade task in which expected reward gain was different depending on the position of the target. Saccadic reaction times were reliably shorter on large-reward trials than on small-reward trials. When position-reward contingency was switched, the reaction time difference changed rapidly. Injecting dopamine D1 antagonist into the caudate significantly attenuated the reward-dependent saccadic reaction time changes. Conversely, injecting D2 antagonist into the same region enhanced the reward-dependent changes. These results suggest that reward-dependent changes in saccadic eye movements depend partly on dopaminergic modulation of neuronal activity in the caudate nucleus. PMID- 16707789 TI - GABAergic network activation of glial cells underlies hippocampal heterosynaptic depression. AB - Tetanus-induced heterosynaptic depression in the hippocampus is a key cellular mechanism in neural networks implicated in learning and memory. A growing body of evidence indicates that glial cells are important modulators of synaptic functions, but very little is known about their role in heterosynaptic plasticity. We examined the role of glial cells in heterosynaptic depression, knowing that tetanization and NMDA application caused depression of synaptic field responses (fEPSPs) and induced Ca2+ rise in glial cells. Here we report that chelating Ca2+ in a glial syncytium interfered with heterosynaptic depression and NMDA-induced fEPSP depression, suggesting that Ca2+ activation of glial cells is necessary for heterosynaptic depression. The NMDA-induced Ca2+ rise in glial cells was sensitive to tetrodotoxin and reduced by the GABAB antagonist CGP55845. Both heterosynaptic depression and simultaneous Ca2+ activation of glial cells were prevented by CGP55845, suggesting an involvement of the GABAergic network in glial activation and heterosynaptic depression. Also, the GABAB agonist baclofen caused both a Ca2+ rise in glial cells and fEPSP depression. Heterosynaptic depression, as well as NMDA- and baclofen-induced depression, were attenuated by an A1 antagonist, cyclopentyl-theophylline, whereas glial cell activation was not, indicating a role of adenosine downstream of glial activation. Finally, heterosynaptic depression requires ATP degradation because ectonucleotidase inhibitors reduced this plasticity. Our work indicates that Ca2+ activation of glial cells is necessary for heterosynaptic depression, which involves the sequential interaction of Schaffer collaterals, the GABAergic network, and glia. Thus, glial and neuronal networks are functionally associated during the genesis of heterosynaptic plasticity at mammalian central excitatory synapses. PMID- 16707790 TI - Necdin promotes GABAergic neuron differentiation in cooperation with Dlx homeodomain proteins. AB - Necdin, a member of the MAGE (melanoma antigen) protein family, is expressed predominantly in terminally differentiated neurons. The necdin gene NDN is maternally imprinted and expressed only from the paternal allele, the deficiency of which is implicated in the pathogenesis of the neurodevelopmental disorder Prader-Willi syndrome. Necdin binds to its homologous MAGE protein MAGE-D1 (also known as NRAGE or Dlxin-1), which interacts with Msx (msh homeobox) and Dlx (distal-less homeobox) family homeodomain transcription factors. Members of the Dlx homeobox gene family are involved in the differentiation and specification of forebrain GABAergic neurons. Here we demonstrate that necdin associates with Dlx homeodomain proteins via MAGE-D1 to promote the differentiation of GABAergic neurons in mouse embryonic forebrain. Immunohistochemical analysis revealed that necdin was coexpressed with Dlx2, Dlx5, or MAGE-D1 in a subpopulation of embryonic forebrain cells. Necdin bound to Dlx2 and Dlx5 via MAGE-D1 and enhanced Dlx2-dependent activation of the Wnt1 (wingless-type MMTV integration site family) promoter. Necdin significantly increased the populations of cells expressing the GABAergic neuron markers calbindin D-28k and glutamic acid decarboxylase when overexpressed by electroporation in cultured forebrain slices. In this assay, Dlx5N, a truncated Dlx5 mutant that competes with Dlx2 to bind MAGE-D1, diminished the effect of necdin on GABAergic neuron differentiation. Furthermore, mutant mice lacking the paternal necdin allele showed a significant reduction in the differentiation of forebrain GABAergic neurons in vivo and in vitro. These results suggest that paternally expressed necdin facilitates the differentiation and specification of GABAergic neurons in cooperation with Dlx homeodomain proteins. PMID- 16707791 TI - Involvement of protein kinase A in patterning of the mouse somatosensory cortex. AB - Patterning of the mouse somatosensory cortex is unusually evident because of the presence of a "barrel field." Presynaptic serotonin and postsynaptic glutamate receptors regulate barrel formation, but little is known of the intracellular signaling pathways through which they act. To determine whether protein kinase A (PKA) plays a role in the development of the barrel field, we examined five viable PKA subunit-specific knock-out (KO) mouse lines for barrel field abnormalities. Barrels are present in these mice, but those lacking the RIIbeta subunit display significantly reduced contrast between the cell densities of barrel hollows and sides compared with wild-type animals. Thalamocortical afferent segregation in the posterior medial barrel subfield appeared normal, suggesting a postsynaptic site of gene action for the RIIbeta protein. Immunoelectron microscopy confirmed that RIIbeta was selectively localized to dendrites and dendritic spines. Mice lacking RIIbeta show reduced glutamate receptor A (GluRA) subunit insertion into the postsynaptic density in postnatal day 7 somatosensory cortex; however, GluRA KO mice developed normal barrels. Our results clearly demonstrate a role for postsynaptic PKA signaling pathways in barrel differentiation. They also demonstrate a clear dissociation between the regulation of GluRA trafficking by PKA and its role in barrel formation. Finally, although a role for PKA downstream of cAMP cannot be ruled out, these data suggest that PKA may not be the principle downstream target because none of the mutants showed a barrelless phenotype similar to that observed in adenylate cyclase type 1 KO mice. These results give insight into activity-dependent mechanisms that regulate barrel formation. PMID- 16707792 TI - Glucocorticoid hormones decrease proliferation of embryonic neural stem cells through ubiquitin-mediated degradation of cyclin D1. AB - Corticosteroids can influence brain function, and glucocorticoid hormone receptors (GRs) are present in brain tissue. We observed that GR and also mineralocorticoid receptor (MR) are expressed by embryonic rat neural stem cells (NSCs). NSCs in developing ventricular epithelium were positive for GR. Stimulation of cultured NSCs with the specific receptor ligands dexamethasone and corticosterone reduced cell proliferation, shown by 5'-bromo-2-deoxy-uridine labeling. The effect of the hormones was dose dependent and inhibited by the GR blocker mifepristone but not by spironolactone, blocking MR. Dexamethasone inhibited the cell cycle by decreasing the levels of cyclin D1 in NSCs. The hormone-induced decline was inhibited by MG132 (benzyloxycarbonyl-leucyl-leucyl leucinal), showing an involvement of the ubiquitin proteasome system, In keeping with this, dexamethasone increased the ubiquitination of cyclin D1. In embryonic brain, dexamethasone inhibited cell proliferation of NSCs. This demonstrates that embryonic NSCs are critically influenced by glucocorticoids, which can have long term effects in the brain. PMID- 16707793 TI - The anion-selective pore of the bestrophins, a family of chloride channels associated with retinal degeneration. AB - Mutations in human bestrophin-1 (VMD2) are genetically linked to a juvenile form of macular degeneration and autosomal dominant vitreoretinochoroidopathy. Recently, it has been proposed that bestrophins are Cl- channels and that the putative second transmembrane domain participates in forming the bestrophin pore. However, the structural determinants of Cl- ion permeation through the channel pore are not known. Here we systematically replaced every amino acid in mouse bestrophin-2 (mBest2) between positions 69 and 104 with cysteine. We then measured the effects on the relative permeability and conductance of the channel to Cl- and SCN- (thiocyanate) and determined the accessibility of the cysteine substituted amino acids to extracellularly applied, membrane-impermeant sulfhydryl reagents. Unlike K+ channels, the amino acids forming the mBest2 selectivity filter are not discretely localized but are distributed over approximately 20 amino acids within the transmembrane domain. Cysteine substituted amino acids in the selectivity filter are easily accessible to extracellularly applied sulfhydryl reagents and select for anionic sulfhydryl reagents over cationic ones. Understanding the structure of the anion conduction pathway of bestrophins provides insights into how mutations produce channel dysfunction and may provide important information for development of therapeutic strategies for treating macular degeneration. PMID- 16707794 TI - A novel pressor area at the medullo-cervical junction that is not dependent on the RVLM: efferent pathways and chemical mediators. AB - Chemical stimulation of a region extending from the most caudal ventrolateral medulla into the upper cervical spinal cord evoked large sympathetically mediated pressor responses. These responses were not dependent on the integrity of the rostral ventrolateral medulla (RVLM) and may be mediated by glutamatergic neurons embedded in the white matter that project to the thoracic spinal cord. We term this new region the medullo-cervical pressor area (MCPA). This region is distinct from the caudal pressor area, because blockade of the RVLM with muscimol inhibited this pressor response but not that evoked from the MCPA. This is the first study to provide functional evidence for a cardiovascular role for neurons in the cervical spinal cord white matter that innervate sympathetic preganglionic neurons (Jansen and Loewy, 1997). Using retrograde tracing, in combination with immunohistochemistry and in situ hybridization, we identified two groups of spinally projecting neurons in the region. Approximately 50% of neurons in one group were excitatory because they contained vesicular glutamate transporter 1 (VGluT1)/VGluT2 mRNA, whereas the other contained a mixed population of neurons, some of which contained either VGluT1/VGluT2 or GAD67 (glutamic acid decarboxylase 67) mRNA. Despite the fact that activation of the MCPA causes potent sympathoexcitation, it does not act to restore arterial pressure after chemical lesion of the RVLM so that a role for this novel descending sympathoexcitatory region remains to be elucidated. PMID- 16707795 TI - Dendritic calcium spikes are tunable triggers of cannabinoid release and short term synaptic plasticity in cerebellar Purkinje neurons. AB - Understanding the relationship between dendritic excitability and synaptic plasticity is vital for determining how dendrites regulate the input-output function of the neuron. Dendritic calcium spikes have been associated with the induction of long-term changes in synaptic efficacy. Here we use direct recordings from cerebellar Purkinje cell dendrites to show that synaptically activated local dendritic calcium spikes are potent triggers of cannabinoid release, producing a profound and short-term reduction in synaptic efficacy at parallel fiber synapses. Enhancing dendritic excitability by modulating dendritic large-conductance calcium-activated potassium (BK) channels improves the spread of dendritic calcium spikes and enhances cannabinoid release at the expense of spatial specificity. Our findings reveal that dendritic calcium spikes provide a local and tunable coincidence detection mechanism that readjusts synaptic gain when synchronous activity reaches a threshold, and they reveal a tight link between the regulation of dendritic excitability and the induction of synaptic plasticity. PMID- 16707796 TI - The impact of astrocytic gap junctional coupling on potassium buffering in the hippocampus. AB - Astrocytic gap junctions have been suggested to contribute to spatial buffering of potassium in the brain. Direct evidence has been difficult to gather because of the lack of astrocyte-specific gap junction blockers. We obtained mice with coupling-deficient astrocytes by crossing conditional connexin43-deficient mice with connexin30(-/-) mice. Similar to wild-type astrocytes, genetically uncoupled hippocampal astrocytes displayed negative resting membrane potentials, time- and voltage-independent whole-cell currents, and typical astrocyte morphologies. Astrocyte densities were also unchanged. Using potassium-selective microelectrodes, we assessed changes in potassium buffering in hippocampal slices of mice with coupling-deficient astrocytes. We demonstrate that astrocytic gap junctions accelerate potassium clearance, limit potassium accumulation during synchronized neuronal firing, and aid in radial potassium relocation in the stratum lacunosum moleculare. Furthermore, slices of mice with coupling-deficient astrocytes displayed a reduced threshold for the generation of epileptiform events. However, it was evident that radial relocation of potassium in the stratum radiatum was not dependent on gap junctional coupling. We suggest that the perpendicular array of individual astrocytes in the stratum radiatum makes these cells ideally suited for spatial buffering of potassium released by pyramidal cells, independent of gap junctions. In general, a surprisingly large capacity for K+ clearance was conserved in mice with coupling-deficient astrocytes, indicating that gap junction-dependent processes only partially account for K+ buffering in the hippocampus. PMID- 16707797 TI - Neurophysiology of implicit timing in serial choice reaction-time performance. AB - Neural representations of time for the judgment of temporal durations are reflected in electroencephalographic (EEG) slow brain potentials, as established in time production and perception tasks. Here, we investigated whether anticipatory processes in reaction-time procedures are governed by similar mechanisms of interval timing. We used a choice reaction task with two different, temporally regular stimulus presentation regimes, both with occasional deviant interstimulus intervals. Temporal preparation was shown in the form of adjustments in time course of slow brain potentials, such that they reached their maximum amplitude just before a new trial, independent of the duration of the interstimulus interval. Preparation was focused on a brief time window, demonstrated by a drop in amplitude of slow potentials as the standard interval had elapsed in deviant interstimulus intervals. Implicit timing influencing perceptual processing was shown in reduced visual-evoked responses to delayed stimuli after a deviant interstimulus interval and in a reduction of EEG alpha power over the visual cortex at the time when the standard interval had elapsed. In contrast to explicit timing tasks, the slow brain potential manifestations of implicit timing originated in the lateral instead of the medial premotor cortex. Together, the results show that temporal regularities set up a narrow time window of motor and sensory attention, demonstrating the operation of interval timing in reaction time performance. The divergence in slow brain potential distribution between implicit and explicit timing tasks suggests that interval timing for different behaviors relies on qualitatively similar mechanisms implemented in distinct cortical substrates. PMID- 16707798 TI - Phosphatidylinositol 3 kinase activation and AMPA receptor subunit trafficking underlie the potentiation of miniature EPSC amplitudes triggered by the activation of L-type calcium channels. AB - We have characterized a mechanism by which the amplitudes of miniature EPSCs (mEPSCs) in CA1 pyramidal neurons in rat hippocampal organotypic slice cultures are potentiated by approximately twofold after a series of depolarizing voltage pulses from -80 to +20 mV. The increase in mEPSC amplitudes is triggered by the activation of L-type calcium channels and is independent of NMDA receptor (NMDAR) activation but also requires calcium release from intracellular stores. The potentiation induced by depolarizing pulses does not alter the kinetic parameters of mEPSCs. The induction phase of this potentiation involves phosphatidylinositol 3 kinase (PI3 kinase) activation because it is blocked completely in the presence of the PI3 kinase inhibitors wortmannin and 2-(4-morpholinyl)-8-phenyl-4H-1 benzopyran-4-one (LY294002). Furthermore, we show that the maintenance phase of depolarizing pulse potentiation requires continued PI3 kinase activity because the application of either wortmannin or LY294002 results in a reversal to control levels of the amplitudes of mEPSCs. Finally, we demonstrate that the increase in mEPSC amplitudes is mediated by the increased expression of functional AMPA receptors (AMPARs) because the potentiation is blocked by N-ethylmaleimide, botulinum toxin A, and a variety of short-sequence peptides that disrupt the interaction of AMPAR subunits with proteins involved with the trafficking of these to the cell membrane. Our data are consistent with the notion that PI3 kinase and membrane fusion/trafficking events play a pivotal role in coordinating changes in synaptic strength, mediated by AMPA receptors, which are triggered by alterations in postsynaptic calcium concentrations whether these changes are initiated via NMDAR-dependent or NMDAR-independent routes. PMID- 16707799 TI - Symmetry of cortical folding abnormalities in Williams syndrome revealed by surface-based analyses. AB - We analyzed folding abnormalities in the cerebral cortex of subjects with Williams syndrome (WS), a genetically based developmental disorder, using surface based analyses applied to structural magnetic resonance imaging data. Surfaces generated from each individual hemisphere were registered to a common atlas target (the PALS-B12 atlas). Maps of sulcal depth (distance from the cerebral hull) were combined across individuals to generate maps of average sulcal depth for WS and control subjects, along with depth-difference maps and t-statistic maps that accounted for within-group variability. Significant structural abnormalities were identified in 33 locations, arranged as 16 bilaterally symmetric pairs plus a lateral temporal region in the right hemisphere. Discrete WS folding abnormalities extended across a broad swath from dorsoposterior to ventroanterior regions of each hemisphere, in cortical areas associated with multiple sensory modalities as well as regions implicated in cognitive and emotional behavior. Hemispheric asymmetry in the temporal cortex is reduced in WS compared with control subjects. These findings provide insights regarding possible developmental mechanisms that give rise to folding abnormalities and to the spectrum of behavioral characteristics associated with WS. PMID- 16707800 TI - Context fear learning in the absence of the hippocampus. AB - Lesions of the rodent hippocampus invariably abolish context fear memories formed in the recent past but do not always prevent new learning. To better understand this discrepancy, we thoroughly examined the acquisition of context fear in rats with pretraining excitotoxic lesions of the dorsal hippocampus. In the first experiment, animals received a shock immediately after placement in the context or after variable delays. Immediate shock produced no context fear learning in lesioned rats or controls. In contrast, delayed shock produced robust context fear learning in both groups. The absence of fear with immediate shock occurs because animals need time to form a representation of the context before shock is presented. The fact that it occurs in both sham and lesioned rats suggests that they learn about the context in a similar manner. However, despite learning about the context in the delay condition, lesioned rats did not acquire as much fear as controls. The second experiment showed that this lesion-induced deficit could be overcome by increasing the number of conditioning trials. Lesioned animals learned normally after multiple shocks, regardless of freezing level or trial spacing. The last experiment showed that animals with complete hippocampus lesions could also learn about the context, although the same lesions produced devastating retrograde amnesia. These results demonstrate that alternative systems can acquire context fear but do so less efficiently than the hippocampus. PMID- 16707801 TI - Brain activation during input from mechanoinsensitive versus polymodal C nociceptors. AB - C-nociceptors mediating cutaneous pain in humans can be distinguished in mechano heat-responsive units (CMH) and mechano-insensitive units (CMi). However, if sensitized in damaged tissue, CMi play an important role in inflammatory pain. CMi differ from CMH by higher electrical thresholds and by mediating the axon reflex. Using these properties, we established two stimulation paradigms: (1) transcutaneous stimulation (TCS) of low current density below the CMi threshold and (2) intracutaneous stimulation (ICS) of high current density that excites CMi. This was proven by the quantification of the axon-reflex flare. Applying these stimulation paradigms during functional magnetic resonance imaging, we investigated whether nociceptor stimulation that recruits CMi leads to different cerebral activation than stimuli that do not recruit CMi. Brain activation by CMi was inferred by subtraction. Both stimuli recruited multiple afferents other than CMi, and we expected a common network of regions involved in different aspects of pain perception and motor nocifensive reactions in both stimuli. ICS that additionally recruited CMi should activate regions with low acuity that are involved in pain memory and emotional attribution. Besides a common network of pain in both stimuli, TCS activated the supplementary motor area, motor thalamic nuclei, the ipsilateral insula, and the medial cingulate cortex. These regions contribute to a pain processing loop that coordinates the nocifensive motor reaction. CMi nociceptor activation did not cause relevant activation in this loop and does not seem to play a role in withdrawal. The posterior cingulate cortex was selectively activated by ICS and is apparently important for the processing of inflammatory pain. PMID- 16707802 TI - Urocortin 2-deficient mice exhibit gender-specific alterations in circadian hypothalamus-pituitary-adrenal axis and depressive-like behavior. AB - Gender differences in hypothalamus-pituitary-adrenal (HPA) axis activation and the prevalence of mood disorders are well documented. Urocortin 2, a recently identified member of the corticotropin-releasing factor family, is expressed in discrete neuroendocrine and stress-related nuclei of the rodent CNS. To determine the physiological role of urocortin 2, mice null for urocortin 2 were generated and HPA axis activity, ingestive, and stress-related behaviors and alterations in expression levels of CRF-related ligands and receptors were examined. Here we report that female, but not male, mice lacking urocortin 2 exhibit a significant increase in the basal daily rhythms of ACTH and corticosterone and a significant decrease in fluid intake and depressive-like behavior. The differential phenotype of urocortin 2 deficiency in female and male mice may imply a role for urocortin 2 in these gender differences. PMID- 16707803 TI - Synaptogenesis of the calyx of Held: rapid onset of function and one-to-one morphological innervation. AB - Synaptogenesis during early development is thought to follow a canonical program whereby synapses increase rapidly in number and individual axons multiply innervate nearby targets. Typically, a subset of inputs then out-competes all others through experience-driven processes to establish stable, long-lasting contacts. We investigated the formation of the calyx of Held, probably the largest nerve terminal in the mammalian CNS. Many basic functional and morphological features of calyx growth have not been studied previously, including whether mono-innervation, a hallmark of this system in adult animals, is established early in development. Evoked postsynaptic currents, recorded from neonatal mice between postnatal day 1 (P1) and P4, increased dramatically from 0.14 +/- 0.04 nA at P1 to -6.71 +/- 0.65 nA at P4 with sharp jumps between P2 and P4. These are the first functional assays of these nascent synapses for ages less than P3. AMPA and NMDA receptor-mediated currents were prominent across this age range. Electron microscopy (EM) revealed a concomitant increase, beginning at P2, in the prevalence of postsynaptic densities (16-fold) and adhering contacts (73 fold) by P4. Therefore, both functional and structural data showed that young calyces could form within 2 d, well before the onset of hearing around P8. Convergence of developing calyces onto postsynaptic targets, indicative of competitive processes that precede mono-innervation, was rare (4 of 29) at P4 as assessed using minimal stimulation electrophysiology protocols. Serial EM sectioning through 19 P4 cells further established the paucity (2 of 19) of convergence. These data indicate that calyces of Held follow a noncanonical program to establish targeted innervation that occurs over a rapid time course and precedes auditory experience. PMID- 16707804 TI - An inhibitor of DNA recombination blocks memory consolidation, but not reconsolidation, in context fear conditioning. AB - Genomic recombination requires cutting, processing, and rejoining of DNA by endonucleases, polymerases, and ligases, among other factors. We have proposed that DNA recombination mechanisms may contribute to long-term memory (LTM) formation in the brain. Our previous studies with the nucleoside analog 1-beta-D arabinofuranosylcytosine triphosphate (ara-CTP), a known inhibitor of DNA ligases and polymerases, showed that this agent blocked consolidation of conditioned taste aversion without interfering with short-term memory (STM). However, because polymerases and ligases are also essential for DNA replication, it remained unclear whether the effects of this drug on consolidation were attributable to interference with DNA recombination or neurogenesis. Here we show, using C57BL/6 mice, that ara-CTP specifically blocks consolidation but not STM of context fear conditioning, a task previously shown not to require neurogenesis. The effects of a single systemic dose of cytosine arabinoside (ara-C) on LTM were evident as early as 6 h after training. In addition, although ara-C impaired LTM, it did not impair general locomotor activity nor induce brain neurotoxicity. Importantly, hippocampal, but not insular cortex, infusions of ara-C also blocked consolidation of context fear conditioning. Separate studies revealed that context fear conditioning training significantly induced nonhomologous DNA end joining activity indicative of DNA ligase-dependent recombination in hippocampal, but not cortex, protein extracts. Finally, unlike inhibition of protein synthesis, systemic ara-C did not block reconsolidation of context fear conditioning. Our results support the idea that DNA recombination is a process specific to consolidation that is not involved in the postreactivation editing of memories. PMID- 16707805 TI - Neural lineages of the Drosophila brain: a three-dimensional digital atlas of the pattern of lineage location and projection at the late larval stage. AB - The late larval brain consists of embryonically produced primary neurons forming a deep core cortex, surrounded at the surface by approximately 100 secondary lineages. Each secondary lineage forms a tract (secondary lineage tract) with an invariant and characteristic trajectory. Within the neuropile, tracts of neighboring lineages bundle together to form secondary tract systems. In this paper, we visualized secondary lineages by the global marker BP106 (neurotactin), as well as green fluorescent protein-labeled clones and thereby establish a comprehensive digital atlas of secondary lineages. The information contained in this atlas is the location of the lineage within the cortex, the neuropile compartment contacted by the lineage tract, and the projection pattern of the lineage tract within the neuropile. We have digitally mapped the expression pattern of three genes, sine oculis, period, and engrailed into the lineage atlas. The atlas will enable us and others to analyze the phenotype of mutant clones in the larval brain. Mutant clones can only be interpreted if the corresponding wild-type clone is well characterized, and our lineage atlas, which visualizes all wild-type lineages, will provide this information. Secondly, secondary lineage tracts form a scaffold of connections in the neuropile that foreshadows adult nerve connections. Thus, starting from the larval atlas and proceeding forward through pupal development, one will be able to reconstruct adult brain connectivity at a high level of resolution. Third, the atlas can serve as a repository for genes expressed in lineage-specific patterns. PMID- 16707806 TI - Early life blockade of 5-hydroxytryptamine 1A receptors normalizes sleep and depression-like behavior in adult knock-out mice lacking the serotonin transporter. AB - In serotonin transporter knock-out (5-HTT-/-) mice, extracellular serotonin (5 HT) levels are markedly elevated in the brain, and rapid eye movement sleep (REMS) is enhanced compared with wild-type mice. We hypothesized that such sleep impairment at adulthood results from excessive serotonergic tone during early life. Thus, we assessed whether neonatal treatment with drugs capable of limiting the impact of 5-HT on the brain could normalize sleep patterns in 5-HTT-/- mutants. We found that treatments initiated at postnatal day 5 and continued for 2 weeks with the 5-HT synthesis inhibitor para-chlorophenylalanine, or for 4 weeks with the 5-HT(1A) receptor (5-HT(1A)R) antagonist N-[2-[4-(2-methoxyphenyl) 1-piperazinyl]ethyl]-N-(2-pyridinyl) cyclohexane carboxamide (WAY 100635), induced total or partial recovery of REMS, respectively, in 5-HTT-/- mutants. Early life treatment with WAY 100635 also reversed the depression-like behavior otherwise observed in these mutants. Possible adaptive changes in 5-HT(1A)R after neonatal treatment with WAY 100635 were investigated by measuring 5-HT(1A) binding sites and 5-HT(1A) mRNA in various REMS- and/or depression-related brain areas, as well as 5-HT(1A)R-mediated hypothermia and inhibition of neuronal firing in the dorsal raphe nucleus. None of these characteristics were modified in parallel with REMS recovery, suggesting that 5-HT(1A)Rs involved in wild-type phenotype rescue in 5-HTT-/- mutants are located in other brain areas or in 5 HT(1A)R-unrelated circuits where they could be transiently expressed during development. The reversal of sleep alterations and depression-like behavior after early life blockade of 5-HT(1A)R in 5-HTT-/- mutants might open new perspectives regarding preventive care of sleep and mood disorders resulting from serotonin transporter impairments during development. PMID- 16707807 TI - The cytokine interleukin-6 is sufficient but not necessary to mimic the peripheral conditioning lesion effect on axonal growth. AB - Lesioning the peripheral branch of a dorsal root ganglion (DRG) neuron before injury of the central branch of the same neuron enables spontaneous regeneration of these spinal axons. This effect is cAMP and transcription dependent. Here, we show that the cytokine interleukin-6 (IL-6) is upregulated in DRG neurons after either a conditioning lesion or treatment with dibutyryl-cAMP. In culture, IL-6 allows neurons to grow in the presence of inhibitors of regeneration present in myelin. Importantly, intrathecal delivery of IL-6 to DRG neurons blocks inhibition by myelin both in vitro and in vivo, effectively mimicking the conditioning lesion. Blocking IL-6 signaling has no effect on the ability of cAMP to overcome myelin inhibitors. Consistent with this, IL-6-deficient mice respond to a conditioning lesion as effectively as wild-type mice. We conclude that IL-6 can mimic both the cAMP effect and the conditioning lesion effect but is not an essential component of either response. PMID- 16707809 TI - Somatosensory coding of roughness: the effect of texture adaptation in direct and indirect touch. AB - To examine somatosensory mechanisms contributing to the perception of roughness, subjects examined surfaces with rigorously specified spatial textures under conditions of direct moving contact between the fingertip and the surface (direct touch) and contact through a rigid probe (indirect touch). Subjects were trained to scan the surfaces along a consistent path and with a speed of 2.7 cm/s. With each mode of touch, periods of prolonged inspection of a single adapting surface were followed by shorter periods in which the roughness of multiple test surfaces was reported. Adaptation caused a drop in perceived roughness under conditions of indirect touch, reflecting the reduced sensitivity of vibrotactile mechanisms that are the main recipients of textural information transmitted through the probe. During direct touch, adaptation had no significant effect on the perception of textures with spatial period >200 mum, which are spatially encoded. The results have an important implication for the physiological basis of the spatial code, which is believed to involve somatosensory cortical neurons with highly structured receptive fields: these cortical populations appear to be less susceptible to adaptation than otherwise similar neuronal populations in the visual system. PMID- 16707808 TI - Linear summation of cat motor cortex outputs. AB - Recruitment of movement-related muscle synergies involves the functional linking of motor cortical points. We asked how the outputs of two simultaneously stimulated motor cortical points would interact. To this end, experiments were done in ketamine-anesthetized cats. When prolonged (e.g., 500 ms) trains of intracortical microstimulation were applied in the primary motor cortex, stimulus currents as low as 10-20 microA evoked coordinated movements of the contralateral forelimb. Paw kinematics in three dimensions and the electromyographic (EMG) activity of eight muscles were simultaneously recorded. We show that the EMG outputs of two cortical points simultaneously stimulated are additive. The movements were represented as displacement vectors pointing from initial to final paw position. The displacement vectors resulting from simultaneous stimulation of two cortical points pointed in nearly the same direction as the algebraic resultant vector. Linear summation of outputs was also found when inhibition at one of the cortical points was reduced by GABAA receptor antagonists. A simple principle emerges from these results. Notwithstanding the underlying complex neuronal circuitry, motor cortex outputs combine nearly linearly in terms of movement direction and muscle activation patterns. Importantly, simultaneous activation does not change the nature of the output at each point. An additional implication is that not all possible movements need be explicitly represented in the motor cortex; a large number of different movements may be synthesized from a smaller repertoire. PMID- 16707810 TI - Theorizing about violence: observations from the Economic and Social Research Council's Violence Research Program. AB - The director of the Economic and Social Research Council Violence Research Program (VRP) in the United Kingdom discusses and debates the impacts of the program in the context of contemporary ideas about violence and current U.K. policy and practice in the field. The projects in the program included 2 historical studies and 18 contemporary studies of violence in the home, schools, prisons, neighborhoods, leisure establishments, massage parlors, and on the street. For example, studies focusing on the nighttime economy in U.K. cities, on paramilitary punishment beatings in Northern Ireland, and on violence experienced and perpetrated by girls are discussed here. Five projects addressed gendered violence, and three addressed domestic violence specifically. Lessons from the VRP are drawn out in this article in a personal account. These lessons include the fact that violence is not hidden, that the meanings of violence are gendered, and that people's accounts of violence matter. PMID- 16707811 TI - Putting a stop to domestic violence in the United kingdom: challenges and opportunities. AB - In this article, the director of Women's Aid, the national domestic violence charity in England, reflects on the role of Women's Aid and activism in the development of responses to domestic abuse and in the Economic and Social Research Council Violence Research Program. The article also reflects on the relationship between researchers and domestic violence activist services. PMID- 16707812 TI - Who listens? The voices of domestic violence survivors in service provision in the United Kingdom. AB - This article discusses, in the context of the United Kingdom, service user participation in domestic violence services and how much the voices of domestic violence survivors are heard in policy and service development. If services addressing domestic violence are to continue to develop and to effectively meet abused women's needs, then the views of those using them need to be heeded and acted on. In the UK, these views have been mainly overlooked in the past. Now, however, as in some other countries, domestic violence survivor participation has been addressed to a small extent. This article discusses some of the pioneering techniques being tried out and the sensitivity and difficulties involved, within a context of an understanding of empowerment and the activist movement against gender violence. The article considers the participation of UK women experiencing violence in both shelter and advocacy services, in wider interagency forums and in policy development. PMID- 16707813 TI - A qualitative exploration of the nature of domestic violence in pregnancy. AB - This article presents a small, qualitative study of 16 women who had experienced domestic violence in the previous 12 months. The violence was perpetrated by a current or former partner in all but one case. Ten of the 16 women had experienced domestic violence during their recent pregnancy, of whom four women had also been assaulted in at least one previous pregnancy. Three women had been assaulted by their partners in a previous pregnancy but not during their recent pregnancy, and three had experienced domestic violence outside of pregnancy only. Some women reported increased feelings of insecurity, jealousy, and possessiveness in their partner during their pregnancy. Abuse within the relationship centered around the arrival and care of the new baby: financial worries, the woman's lessening physical and emotional availability during pregnancy, the lack of practical and emotional support from the male partner, and doubts about paternity. PMID- 16707814 TI - Systemic contact dermatitis. PMID- 16707815 TI - Changing trends in hair restoration surgery. AB - Androgenetic alopecia is an important and common cause for baldness. Despite recent advances, the drug therapy of this condition remains unsatisfactory. Surgical hair restoration is the only permanent method of treating this condition. Introduction of recent techniques such as follicular unit transplantation have improved the cosmetic results and patient satisfaction. This article discusses the latest trends in hair restoration surgery. PMID- 16707816 TI - Disulfiram and low nickel diet in the management of hand eczema: a clinical study. AB - BACKGROUND: Hand eczema due to nickel sensitivity is a challenging task for the dermatologist. The average human diet provides sufficient amount of nickel, which acts as a provocating factor in nickel-sensitive individuals. When such patients are treated with steroid or other immunosuppressives, only short-term remission is obtained. This is because unless the dietary intake of nickel is minimized and the existing amount of nickel in the body of the sensitized individual is depleted, long-term remission is unlikely. AIM: To evaluate the efficacy of oral disulfiram, a nickel-chelating agent and low nickel diet (LND) in reducing the clinical symptoms and preventing frequent relapse of hand eczema in nickel sensitive individuals. METHODS: A total of 21 patients with chronic vesicular hand eczema with nickel sensitivity were taken for this study. Patients were randomly divided into two groups: (a) Study group consisting of 11 patients (8 females and 3 males). They were prescribed disulfiram orally for a period of 4 weeks; they started LND 2 weeks prior to initiation of disulfiram therapy and continued till the end of follow-up period. (b) Control (placebo) group consisting of 10 patients (7 females and 3 males). They were allowed to continue with normal diet. Each of them received lactose tablet daily as placebo for 4 weeks. It was a comparative study and participants were not aware if they belonged to study group or control group (single blind trial). RESULTS: Hand eczema healed completely in 10 (90.9%) out of 11 patients treated with disulfiram and LND during the treatment period in the study group, compared with 1 out 10 patients in control (placebo) group (non significant). Mild relapse was noted in 5 patients in between 2-12 weeks of follow-up period. CONCLUSION: Low nickel diet and short course of oral disulfiram therapy can be considered a good option for the control of chronic hand eczema in nickel-sensitive individuals. PMID- 16707817 TI - Cutaneous manifestations in patients with chronic renal failure on hemodialysis. AB - BACKGROUND: Chronic renal failure (CRF) presents with an array of cutaneous manifestations. Newer changes are being described since the advent of hemodialysis, which prolongs the life expectancy, giving time for these changes to manifest. AIM: The aim of this study was to evaluate the prevalence of dermatologic problems among patients with chronic renal failure (CRF) undergoing hemodialysis. METHODS: One hundred patients with CRF on hemodialysis were examined for cutaneous changes. RESULTS: Eighty-two per cent patients complained of some skin problem. However, on examination, all patients had at least one skin lesion attributable to CRF. The most prevalent finding was xerosis (79%), followed by pallor (60%), pruritus (53%) and cutaneous pigmentation (43%). Other cutaneous manifestations included Kyrle's disease (21%); fungal (30%), bacterial (13%) and viral (12%) infections; uremic frost (3%); purpura (9%); gynecomastia (1%); and dermatitis (2%). The nail changes included half and half nail (21%), koilonychia (18%), onychomycosis (19%), subungual hyperkeratosis (12%), onycholysis (10%), splinter hemorrhages (5%), Mees' lines (7%), Muehrcke's lines (5%) and Beau's lines (2%). Hair changes included sparse body hair (30%), sparse scalp hair (11%) and brittle and lusterless hair (16%). Oral changes included macroglossia with teeth markings (35%), xerostomia (31%), ulcerative stomatitis (29%), angular cheilitis (12%) and uremic breath (8%). Some rare manifestations of CRF like uremic frost, gynecomastia and pseudo-Kaposi's sarcoma were also observed. CONCLUSIONS: CRF is associated with a complex array of cutaneous manifestations caused either by the disease or by treatment. The commonest are xerosis and pruritus and the early recognition of cutaneous signs can relieve suffering and decrease morbidity. PMID- 16707819 TI - Pattern of skin diseases among migrant construction workers in Mangalore. AB - BACKGROUND: With increasing industrialization, the construction industry provides employment to a large number of skilled and nonskilled workers, many of them migrant workers. AIM: This study was undertaken to provide epidemiological data regarding various dermatoses among migrant construction workers in India as currently there is a paucity of the same. METHODS: One thousand construction workers, including 467 migrant laborers, were examined for various dermatoses. RESULTS: Most (88%) workers were males and 51.17% were in their third decade. Infective and noninfective dermatoses were seen in 89.72% and 53.74% of laborers respectively. Among infective dermatoses, fungal infections were the most common (46.25%) ones, followed by bacterial infections (24.83%), scabies (8.56%) and viral infections (6.42%). Contact dermatitis to cement was seen in 12.48% of the laborers. Masons had a significantly higher incidence of contact dermatitis to cement, viral infections and scabies than helpers. CONCLUSIONS: The pattern of dermatoses is an expression of poverty, overcrowding and the occupational hazards of the construction industry. PMID- 16707818 TI - Prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in community acquired primary pyoderma. AB - BACKGROUND: Although prevalence of MRSA strains is reported to be increasing, there are no studies of their prevalence in community-acquired primary pyodermas in western India. AIMS: This study aimed at determining the prevalence of MRSA infection in community-acquired primary pyodermas. METHODS: Open, prospective survey carried out in a tertiary care hospital in Mumbai. MATERIALS AND METHODS: Eighty-six patients with primary pyoderma, visiting the dermatology outpatient, were studied clinically and microbiologically. Sensitivity testing was done for vancomycin, sisomycin, gentamicin, framycetin, erythromycin, methicillin, cefazolin, cefuroxime, penicillin G and ciprofloxacin. Phage typing was done for MRSA positive strains. RESULTS: The culture positivity rate was 83.7%. Staphylococcus aureus was isolated in all cases except two. Barring one, all strains of Staphylococcus were sensitive to methicillin. CONCLUSIONS: Methicillin resistance is uncommon in community-acquired primary pyodermas in Mumbai. Treatment with antibacterials active against MRSA is probably unwarranted for community-acquired primary pyodermas. PMID- 16707820 TI - Chimeric monoclonal antibody to tumor necrosis factor alpha (infliximab) in psoriasis. AB - BACKGROUND: Insights into the pathogenesis of psoriasis have provided opportunities to target key steps in the disease process. Tumor necrosis factor alpha (TNF- alpha) being crucial to the pathogenesis of psoriasis, monoclonal antibodies against this cytokine have proved useful in its treatment. AIM: To study the efficacy of chimeric monoclonal antibody to TNF- alpha (infliximab) in Indian patients with recalcitrant psoriasis vulgaris. MATERIALS AND METHODS: Three patients with recalcitrant psoriasis vulgaris were studied. Baseline haemogram, biochemical parameters, chest radiograph and Mantoux skin test were performed. A loading dose regimen of 5 mg/kg infliximab was administered at weeks 0, 2 and 6. PASI assessment, adverse drug event monitoring and laboratory assessments were carried out at 2-week intervals until week 10. Patients were followed up until week 22 for relapse. RESULTS: Infliximab was well tolerated. The mean PASI was 25.4 at presentation and declined to 5.5 at 10 weeks. PASI 75 was attained at a mean of 9.6 weeks. Relapse occurred at a mean of 18.6 weeks after the first infusion. CONCLUSIONS: This study on Indian patients brings out the importance of cytokine-based therapies in psoriasis. Indigenous production could make these therapies a viable therapeutic option for psoriasis patients in the near future. PMID- 16707821 TI - Ichthyosis follicularis with alopecia and photophobia (IFAP) syndrome. AB - A 12-year-old boy born of a nonconsanguineous marriage presented with dry rough skin and photophobia since birth. His growth and developmental milestones were normal and there was no history of any neurological problem, hearing deficit or scarring around the hair follicles. Cutaneous examination revealed diffuse thinning of scalp hair with loss of eyebrows and eyelashes and a sandpapery texture of the skin all over the body, suggestive of ichthyosis follicularis with alopecia and photophobia syndrome. PMID- 16707822 TI - Epidermolytic hyperkeratosis with rickets. AB - A 6-year-old child presented with generalized hyperkeratosis, most marked over the flexures; windswept deformity of the legs; and limping since 3 years. On the basis of the clinical, histopathologic and biochemical findings, he was diagnosed as a case of epidermolytic hyperkeratosis with rickets. He was treated with parenteral vitamin D3 and calcium supplements orally. Nutritional rickets has been reported in children with various types of ichthyosis like lamellar and X linked types. We report this case of epidermolytic hyperkeratosis with rickets for its rarity. PMID- 16707824 TI - Giant cell tumor of skin. AB - Giant cell tumor of the skin is a rare entity showing gross and histological features similar to those of giant cell tumor of the bone. We report a case of malignant giant cell tumor of the thigh in a 55-year-old man. Histological features showed a biphasic population of mononuclear cells admixed with osteoclast-like giant cells. The nuclei of the giant cells were similar to those of the mononuclear cells. This tumor should be distinguished from a variety of cutaneous neoplasms that contain multinucleated giant cells. PMID- 16707823 TI - Mycetoma sans sinuses. AB - Mycetoma is a chronic suppurative infection involving the skin, subcutaneous tissue and bone. The hallmark of mycetoma is tumefaction, draining sinuses and the presence of microcolonies called grains. Sinuses develop in all patients with mycetoma within a year of the appearance of the lesion. The following case is reported as no sinuses had developed despite the presence of the lesion for 9 years, posing a diagnostic dilemma. PMID- 16707825 TI - Hurler syndrome with a tuft of hair. AB - A 2-year-old girl presented with coarse, thick hairy skin all over the body, a tuft of hair in the parietal region, coarse facial features and a prominent forehead with a large tongue, hepatosplenomegaly and skeletal deformities. Mucopolysaccharides excretion spot test of the urine was positive; and an assay for glycosaminoglycans in the urine was also high, which confirmed the clinical diagnosis of Hurler syndrome. We present this rare case to discuss the possibility of the association of mental retardation with a tuft of hair in this syndrome. PMID- 16707826 TI - Nickel nuisance: a clinical observation. PMID- 16707827 TI - Final push of leprosy: it is prudent to pause before declaration! PMID- 16707828 TI - Modified combined methotrexate PUVA therapy in the treatment of recalcitrant psoriasis: a preliminary report. PMID- 16707829 TI - Diagnosis of delayed pressure urticaria. PMID- 16707830 TI - Efficacy of steroid oral mini-pulse therapy in lichen planus: an open trial in 35 patients. PMID- 16707831 TI - Localised bullous pemphigoid in the breast. PMID- 16707832 TI - Split-thickness skin grafting: a novel approach in the treatment of Fournier's gangrene. PMID- 16707833 TI - Scaly signs in dermatology. PMID- 16707834 TI - EndNote 7.0. AB - EndNote is a useful software for online literature search and efficient bibliography management. It helps to format the bibliography according to the citation style of each journal. EndNote stores references in a library file, which can be shared with others. It can connect to online resources like PubMed and retrieve search results as per the search criteria. It can also effortlessly integrate with popular word processors like MS Word. The Indian Journal of Dermatology, Venereology and Leprology website has a provision to import references to EndNote. PMID- 16707835 TI - Acitretin. PMID- 16707836 TI - Anti-CD20 monoclonal antibody (rituximab) in the treatment of pemphigus. PMID- 16707837 TI - Pericardial effusion with vesiculobullous lesions in a young female. Bullous systemic lupus erythematosus (bullous SLE). PMID- 16707838 TI - Periumbilical keratotic papules. PMID- 16707839 TI - Actinomycotic mycetoma of the hand: A rare occurrence. PMID- 16707840 TI - Acquired smooth muscle hamartoma. AB - Smooth muscle hamartoma is an uncommon, usually congenital, cutaneous hyperplasia of the arrectores pilorum muscles. When it is acquired, it may be confused with Becker's nevus. We report a case of this rare tumor in a 19-year-old man. The disease started several years ago as multiple small skin-colored papules that subsequently coalesced to form a large soft plaque on the back of the left shoulder. The diagnosis of acquired smooth muscle hamartoma was confirmed on histopathology. The patient was reassured about the benign nature of the lesion and was not advised any treatment. PMID- 16707841 TI - A patch of alopecia on chin. Alopecia mucinosa (follicular mucinosis). PMID- 16707842 TI - Prostaglandin facilitates afferent nerve activity via EP1 receptors during urinary bladder inflammation in rats. AB - We examined the effects of loxoprofen, a cyclooxygenase inhibitor, and ONO-8711, an EP1-receptor antagonist, on afferent nerve activity during acetic acid (AA, 0.1% v/v)-induced inflammation of the rat urinary bladder. Distension stimulation was performed (vesical pressure of 30 cm H2O) for 2 min. The neuronal discharge was recorded from L6 dorsal root filaments. The discharge was observed just after the beginning of distension and increased gradually thereafter. When the vesical pressure returned to control value, the discharge diminished abruptly. AA infusion increased the total number of spikes to 198 +/- 38.8% of control values. AA infusion also produced asynchronous discharge in 39% of the animals. Loxoprofen administration (1 mg/kg, i.v.) reduced the number of spikes to 40.3 +/ 14.8% of control values. ONO-8711 administration (1 and 3 mg/kg, i.v.) reduced the number of spikes to 81.6 +/- 1.6% and 32.2 +/- 7.4% of control values, respectively. These data indicate that loxoprofen or EP1-receptor antagonist inhibit the inflammation-related neuronal activity. EP1 receptors in the peripheral afferent nerve terminal and/or urothelium may facilitate the primary afferent nerve activity, which elicits the inflammation-induced micturition reflex. PMID- 16707843 TI - Dynamic movement of center of gravity with hand grip. AB - We studied the movement of center of gravity (CG) in young and aged subjects during maximum grip of right or left hand. Body-sway was recorded with stabilometry in rest-stand position and in maximum grip. The data from right-or left-handed young subjects were analyzed. Maximum grip power was not different between dominant hand and un-dominant hands. Total length (LNG) and total movement area of CG (REC AREA) during the measurements were significantly larger in maximum grip than in rest-stand. In right-handed subjects, LNG increased to 245% and 250% of rest-stand value, and REC AREA increased to 589% and 633% in right and left hand grip, respectively. In left-handed subjects, LNG increased to 186% and 188% of rest-stand value, and REC AREA increased to 400% and 533% in right and left hand grip, respectively. No significant difference of LNG and REC AREA was observed between right and left hand grip in either hand dominant subject Maximum grip did not affect CG in rest-stand. In aged subjects, maximum grip power was significantly less than in young subjects (48%). LNG and REC AREA in rest-stand were significantly larger in aged subjects than in young subjects (220% and 400%, respectively). They were not different during maximum grip with either hand. While aged subjects have difficulty of controlling CG in rest-stand, they have less problems to stabilize CG during maximum grip. These data indicated that dynamic movement of CG might be important to understand person's activity of daily living. PMID- 16707844 TI - Possible activation of murine T lymphocyte through CD98 is independent of interleukin 2/interleukin 2 receptor system. AB - CD98 is a widely expressed cell surface heterodimeric protein of 125 kDa. Its expression is upregulated during lymphocyte activation induced by mitogen, superantigen, conventional antigen, and a combination of phorbol myristate acetate (PMA) and ionomycin. However, the role of CD98 in the immune system is not so well understood. The role of CD98 in murine T lymphocyte proliferation was investigated, especially in correlation with the interleukin 2 (IL-2)/interleukin 2 receptor (IL-2R) system. Monoclonal antibody (mAb) directed against murine CD98 heavy chain (mCD98HC) suppressed the proliferation of lymphocytes stimulated with concanavalin A (Con A). Anti-mCD98HC mAb did not suppress the expression of IL 2Ralpha. Anti-IL-2Ralpha mAb, which suppressed DNA synthesis, did not inhibit the expression of CD98HC. Murine IL-2 (recombinant), which induced considerable DNA synthesis by lymphocytes stimulated with a sub-optimal dose of Con A, did not induce CD98HC expression in lymphocytes. In addition, anti-mCD98HC mAb did not inhibit the production of IL-2 by lymphocyte stimulated with Con A. Taken together with these findings, it was speculated that the CD98 system is independent of the IL-2/IL-2R system in murine T lymphocyte activation. PMID- 16707845 TI - Inhibitory action of eugenol compounds on the production of nitric oxide in RAW264.7 macrophages. AB - Effects of eugenol compounds on the production of nitric oxide (NO) in RAW264.7 macrophages were analyzed in relation to the anti-inflammatory action of these compounds. Eugenol and isoeugenol inhibited lipopolysaccharide (LPS)-dependent production of NO, which was due to the inhibition of protein synthesis of inducible nitric oxide synthase (iNOS). Isoeugenol showed the most effective inhibitory effect and eugenol was less effective. LPS-dependent expression of cyclooxygenase-2 (COX-2) protein was also inhibited markedly by isoeugenol, and less effectively by eugenol. Anti-inflammatory action of eugenol compounds may be explained by the inhibition of NO production and COX-2 expression, the pro inflammatory mediators. PMID- 16707846 TI - Effect of retinoic acid-induced transglutaminase on cell growth in regenerating liver. AB - Transglutaminase 2 (TG2) is implicated in the inhibitory regulation of the hepatocyte growth in vitro. In vivo, however, the role of TG2 in liver regeneration after partial hepatectomy (PH) is almost unknown. A dramatic increase of TG2 expression and activation is induced by retinoic acid (RA). Here we show the effect of the RA-induced overexpression of TG2 on liver regeneration after PH. Regenerating rat liver was prepared by 70% PH. RA was intraperitoneally injected immediately after PH. TG2 activity was determined by incorporation of 14C-putrescine into dimethylcasein. Cell cycle was evaluated for incorporation of BrdU into hepatocytes and detected by a flow cytometric analysis. The treatment of RA greatly increased TG2 activity at 1 day after PH. At that time, DNA synthesis was significantly reduced by the treatment of RA. The recovery of liver weight after PH was significantly delayed by the treatment of RA. These results suggested that TG2 was involved in growth capacity in regenerating rat liver after PH. PMID- 16707847 TI - Continuous carbachol infusion promotes peripheral cell proliferation and mimics vagus hyperactivity in a rat model of hypothalamic obesity. AB - Lesions of the ventromedial hypothalamus (VMH) result in obesity and enhanced cellular proliferation in various organs, including the pancreas, gastrointestinal tract, and liver. Previous studies have suggested that vagal hyperactivity, rather than overeating, induces the peripheral cell proliferation in VMH-lesioned rats. The goal of the present study was to investigate the mechanism of peripheral cell proliferation in VMH-lesion-induced obesity by infusing rats with the acetylcholine agonist, carbachol, and then measuring cellular proliferation in the pancreas and duodenum using immunohistochemistry. The ventromedial hypothalamus was bilaterally lesioned in five rats. In other rats, the bilateral vagus nerves were ligated (vagotomized), and saline or carbachol was continuously administered by an osmotic minipump (n = 5 in each group). Three days later, rats were killed, and cell proliferation was assessed in the pancreas and the duodenum using immunohistochemistry for proliferating cell nuclear antigen (PCNA). Additionally, cellular proliferation in the duodenum was more precisely examined by assessing incorporation of 5-bromo-2'-deoxyuridine (BrdU). Cellular proliferation was higher in rats that received carbachol infusions and in rats with VMH-lesions when compared with control rats (P < 0.05, respectively). The pancreatic PCNA-expressing cells were predominantly identified as the B-cells of the islets of Langerhans. These data demonstrate that carbachol infusion can induce pancreatic and duodenal cell proliferation to a degree that was comparable to that in vagal hyperactivity induced by VMH lesions. PMID- 16707848 TI - 1,25-Dihydroxyvitamin D3 suppresses exportin expression in human promyelocytic leukemia HL-60 cells. AB - The active form of vitamin D3, 1alpha,25-dihydroxyvitamin D3 (DVD), is a potent inducer of cell differentiation and inhibits proliferation of cells such as human promyelocytic leukemia HL-60 cells. In the present study, we examined the effects of DVD on the expression of exportin-1 and exportin-t, which play essential roles in the transport of mRNA and tRNA, respectively. The results of reverse transcription-polymerase chain reaction and quantitative real-time polymerase chain reaction indicated that DVD down-regulated the gene expression of these exportins. Western blotting revealed the decreased production of these proteins in DVD-treated cells. Thus, the present findings of decreased expression of exportin-1 and exportin-t induced by DVD can be correlated to inhibition of the proliferation of HL-60 cells. PMID- 16707849 TI - Airway remodeling in asthma and its influence on clinical pathophysiology. AB - Bronchial asthma has been characterized by chronic and allergic airway inflammation, which induces cytological and histological changes in the airway structure over time. These changes have been called airway remodeling, which includes goblet cell hyperplasia, subepithelial fibrosis, and hyperplasia and hypertrophy of airway smooth muscle cells. Airway epithelium in asthma is often occupied with goblet cells, which contain secretory granules. Airway wall thickness increases because of subepithelial fibrosis, and hyperplasia and hypertrophy of airway smooth muscle cells and submucosal glands. Airway remodeling, therefore, can often cause irreversible airflow limitation, an increase of airway hyperresponsiveness and severity of asthma. Recent studies have demonstrated the molecular and cellular mechanisms of goblet cell hyperplasia, subepithelial fibrosis, and hyperplasia and hypertrophy of airway smooth muscle cells. Several lines of evidence suggest that airway remodeling has been induced by cytokines and mediators produced in chronic allergic airway inflammation. Thus, early intervention with inhaled corticosteroid may prevent progress of airway remodeling by suppressing allergic airway inflammation. PMID- 16707850 TI - Leupeptin, a calpain inhibitor, protects inner ear hair cells from aminoglycoside ototoxicity. AB - Inner ear hair cells play a major role in the auditory pathway that converts sound stimulation into electrical signals, and then into a neural code. However this function is often lost by aminoglycoside ototoxicity. The injury of inner ear hair cells from aminoglycoside treatment is considered apoptosis, and caspase is an important participant in the apoptosis pathway in many organs. It has been reported that calpain, a calcium-dependent protease, is essential for mediation and promotion of cell death. The purpose of the present study was to investigate effects of caspase and calpain inhibitors on the inner ear hair cells after aminoglycoside treatment, and to explore the cell death pathway. Cochlea explant cultures were prepared from mice of postnatal 6 days, cultured with neomycin and/or protease inhibitors, and then stained with phalloidin-fluorescein isothiocyanate (phalloidin-FITC), which was used as a marker to identify surviving hair cells. We demonstrated that neomycin (0.1-1 mM) reduced the number of outer hair cells in a dose-dependent manner. Furthermore, we showed that leupeptin, a calpain inhibitor, significantly protects against the neomycin induced loss of outer hair cells, whereas a caspase inhibitor was effective only against a lower concentration of neomycin (0.2 mM). Using the TdT-mediated dUTP biotin nick and labeling method, we also found that a calpain inhibitor, but not a caspase inhibitor, prevents apoptotic DNA fragmentation after treatment with 1 mM neomycin. These results suggest that calpain, rather than caspase, may be responsible for apoptosis induced by aminoglycoside. Thus, leupeptin may prevent hearing loss from aminoglycoside ototoxity. PMID- 16707851 TI - Captopril, an Angiotensin-converting enzyme inhibitor, attenuates the severity of acute pancreatitis in rats by reducing expression of matrix metalloproteinase 9. AB - It has been reported that matrix metalloproteinase 9 (MMP-9) disrupts basement membrane and increases vascular permeability. MMP-9 therefore might participate in the pathogenesis of severe acute pancreatitis (SAP). Captopril, an angiotensin converting enzyme inhibitor, could reduce MMP-9 expression. However, the effect of captopril on the outcome of SAP is not ascertained. The aim of this study was to determine whether captopril attenuates the severity of SAP by reducing MMP-9 expression. Thirty Sprague-Dawley rats were randomly divided into 3 groups (n = 10 for each). Rats were given intraperitoneal injection of saline (SAP group) or captopril (4 mg/kg) (treated group), and then given retrograde infusion of 5% sodium taurocholate (1.5 ml/kg) into the pancreatic duct under laparotomy to induce SAP. One group of rats, injected with saline, underwent only sham operation (Control). Experimental samples were collected at 24 hrs after the induction of SAP or sham operation. Various markers of severity of SAP, such as serum levels of amylase and trypsinogen activation peptide and the vascular permeability, were increased in rats with SAP, but were significantly decreased in captopril-treated rats (p < 0.01). Likewise, the serum MMP-9 levels and expression levels of pancreatic tissue MMP-9 were significantly higher in rats with SAP than those in captopril-treated rats and control rats (p < 0.01 for both parameters), but showed no difference between captopril-treated and control rats. These results suggest that captopril may attenuate vascular permeability by reducing MMP-9 expression in SAP, thereby ameliorating severity of the disease. The use of captopril might become a new therapeutic agent for SAP. PMID- 16707852 TI - Expression of myoferlin in skeletal muscles of patients with dysferlinopathy. AB - Myoferlin is a novel protein of unknown function with high homology to dysferlin, the gene mutations of which cause limb girdle muscular dystrophy type 2B and Miyoshi myopathy. The myoferlin gene seems to be a candidate for the modifier, and because of the high homology to dysferlin myoferlin may work as a compensator for the absence of dysferlin in dysferlinopathy. This hypothesis is based on the observation that utrophin, which has 80% homology with dystrophin, is overexpressing in the dystrophin deficient myofibers. To test this hypothesis, we investigated the myoferlin expression by immunoblot and immunohistochemical analysis in muscles of five patients with dysferlinopathy. For this aim, we generated a myoferlin specific antibody that does not cross react with dysferlin, and performed the immunoblot, immunohistochemical and immunoelectron microscopic studies. Immunohistochemical analysis showed that the antibodies against myoferlin and dysferlin clearly stained the normal human myofiber surface membranes. The electron microscopy of single immunogold labeled samples for myoferlin showed the presence of the molecular signal along the normal muscle cell membrane. Immunoblot analysis showed that the intensity of 230-kDa myoferlin band of dysferlinopathy muscle extracts was similar to that of normal muscle extracts. The immunostaining of dysferlinopathy muscles with anti-myoferlin antibody revealed a weak immunoreactivity along the muscle cell surface. Thus, the compensatory overexpression of myoferlin was not detected in muscles with dysferlinopathy. PMID- 16707853 TI - Hybrid functional electrical stimulation with medial linkage knee-ankle-foot orthoses in complete paraplegics. AB - We have previously restored ambulation in paraplegics by performing hybrid functional electrical stimulation (FES) with medial linkage knee-ankle-foot orthosis (MLKAFO). The most common MLKAFO (hinge-type MLKAFO) has the hypothetical axis that is lower than the physiological hip joint position, resulting in slow velocity and short step length. A new MLKAFO (sliding-type MLKAFO), which uses sliding medial linkages, has been developed to correct the axial discrepancy of the hinge-type MLKAFO that causes limited hip joint excursion. There have been reports of instability associated with sliding medial linkages, but the mechanism of this instability is unclear. The purpose of the present study was to evaluate the effects of FES with MLKAFOs on ambulation in paraplegics. Two complete paraplegic patients (levels T8 and T12, respectively) participated in this study. Kinematics data during ambulation were obtained using a motion analysis system. We measured gait velocity and hip progression during the standing phase. The sliding-type MLKAFO produced faster gait velocity than did the hinge-type MLKAFO, but it caused pelvis instability without FES. Pelvis instability was controlled by hybrid FES using the sliding-type MLKAFO. With hybrid FES, the sliding-type MLKAFO provides better gait performance than the hinge-type MLKAFO, but the hinge-type MLKAFO provides greater pelvis stability during walking. Moreover, FES provides sufficient propulsion to allow the complete paraplegics to walk. PMID- 16707854 TI - Mice with blunted hypoxic ventilatory response are susceptible to respiratory disturbance during hypoxia. AB - Hypoxia causes a life-threatening situation, and the ventilatory response to hypoxia plays an important role in preventing death. We have hypothesized that persons with a blunted hypoxic ventilatory response may have a weak defense response to hypoxic episodes and be susceptible to fatal respiratory disturbances. However, precise correlations between the hypoxic ventilatory response and respiratory disturbances are not well understood. In the present study we examined the hypoxic and hypercapnic ventilatory responses in nine inbred mouse strains (A/J, AKR/N, BALB/c, C3H/He, C57BL/6, DBA/2, NZW, SWR/J, and 129Sv). Breathing frequency, tidal volume and minute ventilation of unanesthetized and unrestrained mice were assessed by whole body plethysmography. Age-matched mice were exposed for 3 min to 10% O(2) in N(2) gas or 10% CO(2) in hyperoxic gas to determine the acute ventilatory response to chemical stimuli. Basal respiratory variables and hypoxic ventilatory responses differed among the strains, but the hypercapnic ventilatory response did not differ. The hypoxic ventilatory response was the highest in AKR/N mice and the lowest in SWR/J mice. These findings suggest that genetic factors may have influenced the hypoxic ventilatory response but not the hypercapnic ventilatory response. To examine the effects of severe hypoxic stress on the respiratory cycle, we exposed the strain with the highest or lowest hypoxic ventilatory response to 6% O(2) in N(2) until the onset of apnea. The "appearance time of apnea", which is defined as the time from the hypoxic loading to the onset of apnea, was shorter in the SWR/J strain than in the AKR/N strain. We suggest that a lower hypoxic ventilatory response may be a risk factor for apnea under hypoxia. PMID- 16707855 TI - Neurological soft signs as the stroke risk in sickle cell disease. AB - Sickle cell disease (SCD) is a common form of hemoglobinopathy and is highly prevalent worldwide. Silent cerebral infarction, which represents infarction without clinical signs, is a risk factor for clinical stroke in patients with SCD. It is well known that silent infarction predisposes patients with SCD to overt stroke. The aim of the present study is to investigate the effect of silent infarction on neurological soft signs (NSS), which demonstrate subtle impairments in sensory integration, motor coordination and the sequencing of complex motor acts and to evaluate whether NSS can be used in clinical practice to evaluate the patients at risk of stroke in SCD patients with silent infarction. Fifty-nine SCD patients without any documented history of cerebrovascular accident and 28 healthy controls were included in this study. All the patients with SCD were evaluated with cerebral magnetic resonance imaging. We found that the NSS scores were significantly higher in patients with silent cerebral infarction than those in patients without silent infarction and control subjects (p < 0.05). Importantly, there was no significant difference in the NSS scores between the patients without silent infarction and control subjects. These results indicate that high NSS scores represnt an important finding for diagnosis of silent infarction in SCD patients. As silent infarction increases the risk for stroke in patients with SCD, NSS can be used to provide additional information in diagnosis of the patients with possible stroke risk during the course of SCD. PMID- 16707857 TI - G-protein beta3 subunit gene variant is unlikely to have a significant influence on serum uric acid level in Japanese workers. AB - The C825T variant of the G-protein beta3 subunit (GNB3) gene has attracted renewed attention as a candidate gene for obesity, hypertension and hyperuricemia. The main role of G-protein is to translate signals from the cell surface into a cellular response. The 825T allele is associated with a splice variant of GNB3 protein and enhanced G-protein activation. We examined the relationship between this variant and the risk of hyperuricemia in Japanese workers. The study subjects were 1,452 men and 1,169 women selected from 3,834 men and 2,591 women in 1997. On the basis of common clinical criteria, hyperuricemia I was defined as serum uric acid >or= 7.0 mg/dl in men and 6.0 mg/dl in women or taking antihyperuricemic medication. The hyperuricemia I group consisted of 186 men and 20 women and its control of 1,266 men and 1,149 women. Hyperuricemia II was defined as serum uric acid > 5.7 mg/dl (median) in men and 3.9 mg/dl (median) in women or taking antihyperuricemic medication. The hyperuricemic II group consisted of 684 men and 570 women and its control of 768 men and 599 women. To replicate previous significant results in young Caucasian men, we selected these criteria because the authors of the study in young Caucasian men adopted the median in their subjects as a cut-off. The statistical power was estimated as 99% based on the significant results in Caucasians. Genotype and allele distributions in men and women with hyperuricemia I and II were not significantly different from those in the corresponding control groups. Logistic regression analysis on hyperuricemia I and II, and multiple regression on serum uric acid level demonstrated no significant effect of the C825T genotype. Despite the sufficient statistical power, this study could not demonstrate the significant influence of C825T on hyperuricemia or serum uric acid. The targeting of this polymorphism is unlikely to be beneficial in the prevention of hyperuricemia in the general Japanese population. PMID- 16707856 TI - Transient reduction of PTI-1 expression by short interfering RNAs inhibits the growth of human prostate cancer cell lines. AB - The prostate tumor-inducing gene 1 (PTI-1) was originally identified by differential ribonucleic acid (RNA) display in human prostate carcinoma. PTI-1 is expressed in human prostate carcinoma but not in benign prostate hypertrophy or normal prostate tissue. PTI-1 may be a member of oncogenes that could affect protein translation and contribute to carcinoma development in human prostate. To investigate the role of PTI-1 in human prostate carcinoma, we constructed three different short interfering RNA (siRNA) vectors (pSilencer3.1-neo-Yu Lei [YL]1-2, -YL3-4 and -YL5-6), each of which was transfected into DU145 and PC3 human prostate cancer cell lines. Among these siRNAs, only pSilencer3.1-neo-YL1-2 could almost completely block the expression of PTI-1 in these two cell lines. The growth of the cell lines was then evaluated after transfection. The proliferation rate was retarded in DU145 and PC3 cells transfected with pSilencer3.1-neo-YL1-2, compared with the cells transfected with a control vector; namely, about 88.6% of DU145 and 80.2% of PC3 cancer cells were blocked at the G1 phase when transfected with pSilencer3.1-neo-YL1-2, compared to 62.0% in DU145 cells and 51.7% in PC3 cells, transfected with the control vector. Moreover, 68.3% of DU145 cells and 72.3% of PC3 cells were induced into apoptosis, while in control transfection, the population was 26.6% in DU145 cells and 28.4% in PC3 cells. These results indicate that blocking PTI-1 expression can inhibit the growth of certain prostate cancer cell lines. We suggest that PTI-1 may serve as a target for the gene-based therapy of human prostate carcinoma. PMID- 16707858 TI - Psychophysiological differences in identical twins discordant for schizophrenia: a critical index for the onset of schizophrenia. AB - It has been hypothesized that not only genetic but also environmental factors contribute to the onset of schizophrenia. We therefore conducted psychophysiological studies on a pair of identical twins discordant for schizophrenia, to differentiate non-genetic from genetic indexes possibly associated with this disease. The affected and unaffected twins were 28 year-old females. The affected twin was diagnosed as having schizophrenia based on the Diagnosis and Statistical Manual of Mental Disorders, third edition revised (DMS III-R), whereas the unaffected twin had no psychiatric disorders. The brain potentials evoked by visual stimulation (visual event-related potential [visual ERP]) were recorded. The components of the visual ERP, which are believed to reflect pattern cognition, semantic processing and the failure to use preceding word information, were compared with normal subjects. Both twins showed abnormal semantic processing and similar failure to use preceding word information. Abnormality of semantic processing was marked in the affected twin. These results indicate that failure to use preceding word information might reflect only genetic factors, whereas abnormal semantic processing might reflect both genetic and non-genetic factors because the affected twin was considered to show accelerated deterioration after the disease onset. However, only the affected twin showed abnormal pattern cognition, which might be attributable to non genetic factors such as an acquired vulnerability to schizophrenia. We suggest that the impairment of pattern cognition evaluated by visual ERP may be a critical index for the onset of schizophrenia. PMID- 16707859 TI - Effective treatment with levodopa and carbidopa for hypomyelination with atrophy of the basal ganglia and cerebellum. AB - Hypomyelination with atrophy of the basal ganglia and cerebellum (H-ABC) is a rare leukoencephalopathy presenting in the infantile period and characterized by diffuse cerebral hypomyelination, and atrophy of the basal ganglia and cerebellum. As patients with H-ABC lack remarkable laboratory findings, the diagnosis is based on brain magnetic resonance imaging findings alone. Only eight cases have been reported in the literature, and thus the natural course and treatment of this disease are not fully understood. We report a 35-month-old boy with H-ABC who had hemidystonia, hypomyelination, and cerebellar ataxia. We diagnosed H-ABC after considering a thorough differential diagnosis, excluding other diseases involving hemidystonia, hypomyelination, and cerebellar ataxia. Furthermore, technetium-99m ethyl cysteinate dimmer-single-photon emission computerized tomography (Tc-ECD-SPECT) and positron emission tomography with fluorodeoxyglucose (18)F (FDG-PET) revealed decreased blood flow and glucose metabolism in the bilateral lenticular nucleus, thalamus, and cerebellum. A peroral levodopa preparation containing carbidopa (levodopa-carbidopa) was effective at ameliorating and stopping the progression of the patient's dystonia (final effective doses: levodopa, 200 mg/day and carbidopa, 20 mg/day). This is the first case report of a Japanese patient with H-ABC and treatment for this disease. Levodopa-carbidopa may be an effective treatment for H-ABC. PMID- 16707860 TI - Inheritance of olfactory preferences I. Pheromone-mediated behavioral responses of Heliothis subflexa x Heliothis virescens hybrid male moths. AB - Shifts in male preference for qualitatively different pheromone blends appear to have played a fundamental role in the divergence of olfactory communication and evolution of moth species. As an initial step in documenting the genetic complexity underlying such shifts, we characterized the behavioral responses of hybrid male moths created by mating two heliothine moth species, Heliothis subflexa and Heliothis virescens. Between 67 and 96% of hybrid males flew upwind and contacted the pheromone source when presented with a blend consisting of (Z) 11-hexadecenal (Z11-16:Ald), (Z)-9-hexadecenal (Z9-16:Ald), and (Z)-11 hexadecenol (Z11-16:OH) in a 1:0.5:0.1 ratio that has previously been shown to be attractive to H. subflexa males. In addition, an H. virescens blend of Z11-16:Ald and (Z)-9-tetradecenal (Z9-14:Ald) enhanced by the addition of Z11-16:OH (in a 1:0.05:0.1 mixture) was attractive to hybrid males (26-64% source contact), but significantly fewer males reached the odor source compared to the blend containing Z9-16:Ald. A blend in which the dosage of Z9-14:Ald was doubled, however, was equally attractive (75-77% source contact) as the Z9-16:Ald containing blend. Consecutive presentation of two blends revealed that individual hybrid males responded equally well to blends containing either Z9-14:Ald or Z9 16:Ald. Together these results suggest that in addition to Z11-16:Ald, hybrid males: (1) required either Z9-16:Ald (likeH. subflexamales) or Z9-14:Ald (like H. virescens males); (2) required the presence of Z11-16:OH (H. subflexa dominant); (3) were not adversely affected by the presence of Z11-16:Ac (H. subflexa dominant). The behavioral response phenotype of hybrid males was therefore influenced by genetic factors inherited from both parental species. PMID- 16707861 TI - Inheritance of olfactory preferences II. Olfactory receptor neuron responses from Heliothis subflexa x Heliothis virescens hybrid male moths. AB - Single-cell electrophysiological recordings were obtained from olfactory receptor neurons (ORNs) in sensilla trichodea on male antennae of hybrids formed mainly by crossing female Heliothis subflexa with male Heliothis virescens ('SV hybrids'). We recorded from the A-, B-, and C-type sensilla trichodea, with the latter two types housing ORNs exhibiting response profiles to different pheromone components that we had previously found to be characteristic for each species. For both the B- and the C-type SV hybrid sensilla, most of the ORNs exhibited a spike amplitude and ORN co-compartmentalization within sensilla that more strongly resembled the ORNs of parental H. subflexa rather than those of H. virescens. The overall mean dose-response profiles of the ORNs in hybrid C- and B-type sensilla were intermediate between those of the H. virescens and H. subflexa parental type ORNs. However, not all hybrid ORNs were intermediate in their tuning spectra, but rather ranged from those that closely resembled H. subflexa or H. virescens parental types to those that were intermediate, even on the same antenna. The most noteworthy shift in ORN responsiveness in hybrid males was an overall increase in sensitivity to Z9-14:Ald exhibited by Z9-16:Ald-responsive ORNs. Heightened cross-responsiveness to Z9-14:Ald by hybrid ORNs correlates well with observed behavioral cross-responsiveness of hybrids in which Z9-14:Ald could substitute for Z9-16:Ald in the pheromone blend, a behavior not observed in parental types. The hybrid ORN shifts involving greater sensitivity to Z9- 14:Ald also correlate well with studies of hybrid male antennal lobe interneurons that exhibited a shift toward greater cross-responsiveness to Z9-14:Ald and Z9- 16:Ald. We propose that the differences between parental H. virescens, H. subflexa, and SV hybrid male pheromone ORN responsiveness to Z9-16:Ald and Z9 14:Ald are most logically explained by an increased or decreased co-expression of two different odorant receptors for each of these compounds on the same ORN. PMID- 16707863 TI - Impact of implantable defibrillators and resynchronization therapy on outcome in patients with left ventricular dysfunction--a meta-analysis. AB - BACKGROUND: The clinical benefits of cardiac resynchronization therapy (CRT) and primary prophylactic implantable cardioverter defibrillator (ICD) in patients with left ventricular systolic dysfunction (LVSD) are debated. OBJECTIVE: To evaluate by a meta-analysis the effect of CRT and prophylactic ICD therapy in patients with LVSD. METHODS: Eligible trials evaluating the effect of CRT vs. no CRT, ICD vs. no-ICD and adding ICD to CRT vs. no-ICD were selected and meta analyzed. The outcomes were: all cause mortality, cardiac mortality, hospitalization for heart failure and change in exercise tolerance and New York Heart Association class. RESULTS: Implantation of CRT reduced all cause mortality odds ratio (OR) = 0.73 (0.60-0.89) p = 0.002 and hospitalization for heart failure OR = 0.60 (0.45, 0.80) p = 0.001, increased peak oxygen consumption by 1.77 (0.32-3.22) ml/kg/min p = 0.017 and improved New York Heart Association class by at least one class with OR = 1.52 (1.30, 1.77) p < 0.0001. Implantation of ICD reduced all-cause mortality OR = 0.75 (0.59-0.96) p = 0.025 and cardiac mortality OR = 0.63 (0.48, 0.82) p = 0.001. Adding ICD to CRT reduced all cause mortality OR = 0.69 (0.53-0.91) p = 0.008. CONCLUSION: Selective patients with LVSD benefit from CRT, ICD or both. Further investigations are necessary to clarify which patients benefit most from a single or combined device implantation. PMID- 16707862 TI - Inheritance of olfactory preferences III. Processing of pheromonal signals in the antennal lobe of Heliothis subflexa x Heliothis virescens hybrid male moths. AB - Pheromone-responsive olfactory interneurons were studied to determine the extent to which their physiological and morphological properties complemented the behavior and peripheral olfactory neurobiology observed in hybrid male moths created by interbreeding two species of heliothine moth, Heliothis virescens and Heliothis subflexa. Complete recordings were made from a total of 33 neurons, and 16 projection neurons (PNs) were subsequently stained with a fluorescent dye. Stained PNs tuned to pheromonal odorants had dendritic arborizations restricted to one of four olfactory glomeruli that together constituted the macroglomerular complex (MGC). As in parental males, PNs tuned to (Z)-11-hexadecenal always had an arbor in the cumulus, the largest of the MGC glomeruli. Previous neurophysiological investigations revealed that PNs with dendritic arbors restricted to the dorso-medial glomerulus (DM) of the MGC responded specifically to either (Z)-9-tetradecenal (Z9-14:Ald; H. virescens males) or (Z)-9-hexadecenal (Z9-16:Ald; H. subflexa males). Hybrid males, which responded equally well in wind tunnel tests to blends containing either Z9-14:Ald or Z9-16:Ald, had DM PNs that responded to both odorants. PNs specific for a third compound, (Z)-11 hexadecenol, required by hybrid males for behavioral activity were localized to the antero-medial MGC glomerulus (AM). Thus, neuronal activity across the cumulus, DM and AM glomeruli represented an attractive blend in hybrid males. Neurons tuned to (Z)-11-hexadecenyl acetate and Z9-14:Ald were restricted to a fourth, ventro-medial glomerulus. The across-glomerular pattern of activity associated with attractive pheromone blends was most similar to that of H. subflexa males, signifying a dominant effect of H. subflexa genes. These results indicate that the behavioral phenotype of hybrid males can be linked to underlying central olfactory characteristics. PMID- 16707864 TI - Influence of different storage methods on laser fluorescence values: a two-year study. AB - The aim of this study was to assess the influence on the infrared laser fluorescence response of some storage methods commonly used in dental research. Forty extracted permanent teeth, selected from a pool of frozen teeth, were divided into four groups of 10. Three groups were stored at 4 degrees C in 1% chloramine, 10% formalin or 0.02% thymol solution. The fourth group was stored at -20 degrees C (no storage solution added). Fluorescence measurements were performed at 14, 77, 113, 168, 232, 486 and 737 days. After 2 years, significant decreases in fluorescence (p<0.01) for the samples in formalin (-60%), chloramine (-72%) and thymol (-54%) were observed. The frozen teeth showed a slight but non significant increase in fluorescence of 5% (p>0.01). Storing solutions have a significant influence on the fluorescence yield. Samples used for in vitro purposes stored frozen do not significantly change their fluorescence response. Thus, cut-off values obtained under the latter conditions could be extrapolated to the in vivo situation. PMID- 16707865 TI - Effect of placing a clear sealant on the validity and reproducibility of occlusal caries detection by a laser fluorescence device: an in vitro study. AB - The principal aim of this in vitro study was to assess the validity and reproducibility of the detection of occlusal caries using laser fluorescence (LF), prior to and following the placement of a clear fissure sealant. It also aimed to compare the manufacturer's standard cut-off recommendations with those published for in vitro studies and to compare the validity and reproducibility of LF with clinical visual examination (CVE) for the detection of occlusal caries under sealants. Three clinicians independently examined visually and with LF 37 extracted teeth (25 molars, 12 premolars), with a range of clinical caries from apparently sound to cavitated dentinal caries. Examinations were conducted under dental surgery conditions. Subsequently, the teeth were serially sectioned to provide the validating criterion. Following placement of the sealant, the specificity generally increased but there was an associated loss of sensitivity, at both the D1 (enamel and dentine) and D3 (dentine) diagnostic thresholds. The LF readings were significantly lower after placement of the sealant (p<0.05). The manufacturer's recommended cut-offs appear to be the most appropriate to use. The CVE had superior validity and reproducibility when compared to LF. Overall, the placement of a clear sealant does influence the detection of caries by LF but does not prevent the detection of caries by this method. PMID- 16707866 TI - Validity and reproducibility of different combinations of methods for occlusal caries detection: an in vitro comparison. AB - This study assessed the validity and reproducibility of different combinations of occlusal caries detection methods: visual examination (VE), laser fluorescence (LF) and radiographic examination (RE). Intra- and interexaminer reproducibilities were also assessed. Forty-seven extracted human molars were used and 121 sites, either suspected or not to be carious, were chosen. Occlusal surfaces were examined by 8 volunteers, assigned to three groups according to their level of knowledge and clinical experience on dental practice: group I, undergraduate students; group II, postgraduate students; group III, professors. Three combinations of methods were tested: A: VE+LF, B: VE+RE, C: VE+LF+RE. The examiners scored the sites using ranking scales and chose a final score based on their clinical experience. The gold standard was determined by histological examination of the sites. In general, LF and RE yielded poorer results than the combinations of methods. For combination A, group III showed the highest sensitivity, while group II showed the highest specificity. For combination B, group II showed moderate sensitivity whereas groups I and III exhibited low sensitivities; all groups of examiners reached substantial specificity. For combination C, all groups exhibited moderate sensitivity and substantial specificity. Interexaminer reproducibility ranged from fair to moderate for combinations A and C, while for combination B kappa values indicated moderate interexaminer reproducibility. It may be concluded that individual exams presented inferior performance than the conjunction of them. Combination C (VE+LF+RE) resulted in the best accuracy for all groups. The knowledge background of the examiners influenced their ability to detect caries lesions and affected interexaminer reproducibility. PMID- 16707867 TI - Accuracy of proximal caries depth measurements: comparison between limited cone beam computed tomography, storage phosphor and film radiography. AB - The aim of this study was to compare the accuracy of limited cone beam computed tomography (LCBCT), an image plate system and F-speed film in assessing the depth of proximal carious lesions. Radiographs of a dry mandible with sound and carious teeth were obtained with all three methods. In 41 molar and premolar proximal surfaces, 2 observers independently measured lesion depth on the images from the three modalities. The correlation of measurements was assessed with Pearson's correlation analysis. Results from imaging modalities and histological sectioning (gold standard) were compared using Bland-Altman plots. Overall comparison of the depth measurements from the imaging modalities and the gold standard was done using repeated-measures ANOVA. Pairwise comparisons of systems were done by the Bonferroni t test. The correlation between the measurements of the two observers was 0.977 for film, 0.997 for image plate system and 0.998 for LCBCT. Bland Altman plots revealed that LCBCT agreed very closely with the gold standard while the agreement between the latter and the image plate--or film--images was moderate. The mean difference and 95% limits of agreement between LCBCT and the gold standard were smaller than those between either image plate or F-speed film and the gold standard. The LCBCT method appears as a promising tool for detection and monitoring of proximal carious lesions. PMID- 16707868 TI - Accuracy and consistency of the visual diagnosis of exposed dentine on worn occlusal/incisal surfaces. AB - Most indices for the assessment of wear of various aetiologies include the distinction between 'enamel still present' and 'dentine exposed' for grading. Since the visual diagnosis of exposed dentine has not yet been validated, the present study is a first attempt to investigate its accuracy and consistency. Sixty-one examiners (23 scientists, 18 university dentists and 20 dental students) were asked to diagnose 49 tooth areas with different grades of wear and to decide whether dentine was exposed (positive test) or not (negative test). Afterwards, the teeth were histologically evaluated. In 44 areas, dentine (also in all cases with minor wear) was exposed, and in 5 areas enamel was present. Overall sensitivity was 0.65, specificity 0.88 and the proportion of correct diagnoses was 0.67. The diagnosis 'dentine is exposed' was about 5 times as likely and the diagnosis 'dentine is not exposed' half as likely to come from an area with exposed dentine than from an enamel-covered area. The closeness of the visual diagnosis to the histological findings was only fair (kappa=0.27), no significant impact of professional experience was found. For inter- and intra examiner agreement, kappa was 0.28 and 0.55, respectively. It was concluded that the diagnosis of exposed dentine is difficult. PMID- 16707869 TI - Exposure time of enamel and dentine to saliva for protection against erosion: a study in vitro. AB - Previous research, mainly on enamel, supports a protective role for salivary pellicle against erosion. Pretreatments have tended to be lengthy (24 h or more) and of questionable relevance to the regular intake of acidic food and drink by many individuals. The aim of this study in vitro was to determine the protective effect of salivary pellicle formed on enamel and dentine over time periods up to 4 h. Flattened, polished human enamel and dentine specimens were pretreated with unstimulated human saliva from a single donor for 2 min, 30 min (enamel only), 1, 2, or 4 h. Controls were exposed to water for the same times. Specimens were then exposed to 0.3% citric acid, pH 3.2 for 10 min with stirring. This cycle was carried out 12 times. Tissue loss was measured by profilometry after 3, 6, 9 and 12 cycles. For enamel, statistically significant protection was found at >or=1 h. For dentine, significant protection was achieved at 2 min. Salivary pellicle offered proportionately greater protection to enamel than dentine. Cautiously extrapolating these in vitro data suggests that pellicle should offer erosion protection to individuals who imbibe acidic drinks at frequencies of 1 h or less. PMID- 16707870 TI - Effect of salivary stimulation on erosion of human and bovine enamel subjected or not to subsequent abrasion: an in situ/ex vivo study. AB - This in situ/ex vivo study evaluated whether saliva stimulated by chewing gum could prevent or reduce the wear and the percent change in microhardness (%SMH) of bovine and human enamel submitted to erosion followed by brushing abrasion immediately or after 1 h. During 2 experimental 7-day crossover phases, 9 previously selected volunteers wore intraoral palatal devices, with 12 enamel specimens (6 human and 6 bovine). In the first phase, the volunteers immersed the device for 5 min in 150 ml of cola drink, 4 times per day (at 8, 12, 16 and 20 h). Immediately after the immersions, no treatment was performed in 4 specimens, 4 other specimens were immediately brushed (0 min) using a fluoride dentifrice, and the device was replaced into the mouth. After 60 min, the remaining 4 specimens were brushed. In the second phase, the procedures were repeated, but after the immersions, the volunteers stimulated the salivary flow rate by chewing a sugar-free gum for 30 min. Changes in wear and %SMH were measured. ANOVA and Tukey's test showed statistical differences (p<0.05) for the following comparisons. The chewing gum promoted less wear and %SMH. A decreasing %SMH and an increasing enamel wear were observed in the following conditions: erosion only, 60 min and 0 min. The human enamel presented greater %SMH and less wear compared to bovine enamel. The data suggest that the salivary stimulation after an erosive or erosive/abrasive attack can reduce the dental wear and the %SMH. PMID- 16707871 TI - Brushing abrasion of eroded bovine enamel pretreated with topical fluorides. AB - Topical fluorides have been proposed for the prevention of erosive dental wear. This study evaluated the in vitro effect of a single professional application of 4% titanium tetrafluoride (TiF4), 1% amine fluoride (AmF) and 0.1% difluorosilane varnish (FV) in preventing wear due to combined erosion and brushing abrasion. One hundred and eight bovine enamel samples were used. Control groups were not pretreated with any product (C), pretreated with a fluoride-free varnish (FV-bl) or pretreated with fluoride varnish and subsequently submitted to varnish removal (FV-r). Wear was modeled by submitting the fluoride-treated and control groups to 3 cycles of the following regimens: erosion/remineralization (er/remin), abrasion/remineralization (abr/remin) or erosion/abrasion/remineralization (er/abr/remin). Erosion was simulated by immersion of the samples for 10 min in citric acid 50 mM (pH 3). Abrasion was carried out for 1 min (200 strokes, load 150 g) in a wear device. Remineralization (2 h artificial saliva) took place between the cycles. Two-way ANOVA showed that there was a significant interaction (p or = 1.5 to < or = 19.9 cm in 78% and > or = 20 cm in 22% of cases, and 64 male subjects, median age 19 years, with 64 BN. In the sample of medium-sized and large CMN, dermoscopic features previously identified as characteristic of congenital lesions (i.e. target network, focal thickening of network lines, target globules, skin furrow hypopigmentation, focal hypopigmentation, hair follicles, perifollicular hypopigmentation, vessels and target vessels) were observed in sufficiently high rates. In the BN group, network, focal hypopigmentation, skin furrow hypopigmentation, hair follicles, perifollicular hypopigmentation and vessels were the main dermoscopic features. Focal thickening of network lines, globules, target globules, homogeneous diffuse pigmentation, hyperpigmented areas, blotches and target vessels were more frequently observed in CMN than in BN. CONCLUSIONS: (1) The same dermoscopic features observed in small and medium-sized CMN by video dermoscopy with high magnifications are also detectable in medium-sized and large CMN, employing the dermoscopy with the 10-fold magnification. (2) Network, focal, skin furrow and perifollicular hypopigmentation, hair follicles and vessels could be considered as peculiar dermoscopic features of BN. (3) Major differences in the frequency of dermoscopic characteristics were detected between CMN and BN, and dermoscopy seems to provide some diagnostic aid in differentiating CMN from BN in equivocal cases. PMID- 16707886 TI - A comparison of the efficacy, relapse rate and side effects among three modalities of systemic corticosteroid therapy for alopecia areata. AB - BACKGROUND: Systemic corticosteroids are one of the most commonly used therapeutic modalities for patients with extensive alopecia areata (AA), although they entail several drawbacks. OBJECTIVE: To determine the best modality for systemic corticosteroid use in terms of their efficacy, relapse rate, and side effects. METHODS: Fifty-one patients with single or multiple AA (AA/multiplex) and 38 patients with alopecia totalis or AA universalis (AA totalis/universalis) were enrolled in this open study. They were randomly divided into three groups depending on the time of their initial visit. They were administered (1) oral dexamethasone (Dex) 0.5 mg/day for 6 months (Dex group), (2) intramuscular triamcinolone acetonide (imTA) 40 mg once a month for 6 months followed by 40 mg once every 1.5 months for 1 year (imTA group), and (3) pulse therapy (PT) using oral predonine 80 mg for 3 consecutive days once every 3 months (PT group). After the treatment, each treatment modality was evaluated by the response rate, relapse rate, and side effect profile. RESULTS: The response rate of AA/multiplex was significantly better in the imTA group than in the Dex group. The overall relapse rate and that of AA totalis/universalis were significantly better in the PT group than in the Dex group. Dysmenorrhea was the most common and problematic side effect. Impairment of the adrenocortical reserve was seen in 7% of the PT group and 23% of the imTA group, which was recovered without any further medical treatment. CONCLUSION: imTA or pulse therapy is effective for AA and has an acceptable level of side effects. The development of a new strategy to reduce the relapse rate is needed. PMID- 16707887 TI - Generalized mucinosis in a patient with erythroderma. AB - We describe an 81-year-old Japanese patient with erythroderma overlapping with widespread and symmetrical deposits of mucin in the upper dermis. Clinically, the mucinous lesions on the nape and upper trunk were localized papular mucinosis. Histologically, there was a perivascular infiltrate of lymphohistiocytic cells mingled with plasma cells in the upper dermis but no sclerosis. Immunohistochemical staining revealed that more than 90% of these infiltrating plasma cells produced immunoglobulin lambda-chain. Both the erythroderma and generalized mucinosis responded to topical steroid and PUVA therapy. To the best of our knowledge, this is the first case of erythroderma accompanied by generalized mucinosis. PMID- 16707888 TI - Wegener's granulomatosis of the penis: genital presentation of systemic disease. AB - Wegener's granulomatosis (WG) is a systemic vasculitis of small and medium-size vessels, mostly affecting the respiratory tract and the kidneys, although any organ may be involved. This case concerns a 50-year-old man in whom WG caused an asymptomatic erosion on the glans penis and prepuce. Histological examination showed an unspecific inflammatory infiltrate. Four months later, a sudden and simultaneous onset of lesions of the upper respiratory tract and kidneys occurred, as well as peripheral neuropathy, polyarthritis, positivity of antineutrophilic cytoplasmic antibodies and its proteinase 3 subset. Histological examination of a nodular lesion on the left leg showed leukocytoclastic vasculitis of small and medium-size vessels, suggesting the diagnosis of WG. Cases in which isolated unspecific erosive lesions of the penis are the only symptoms of WG have been seen in only 4 other patients so far, and a correct diagnosis was only possible due to a careful follow-up study. PMID- 16707890 TI - Successful treatment of nodular actinic reticuloid with tacrolimus ointment. AB - Actinic reticuloid (AR) is the severest clinical variant of chronic actinic dermatitis (CAD) and describes a persistent photoinduced skin disorder often associated with delayed-type hypersensitivity reactions. Histopathologically, AR resembles pseudolymphoma and may also show features of cutaneous lymphoma. In contrast to other UV-induced skin diseases, AR patients show persistent photosensitivity to UV radiation and visible light parallel to permanent changes of skin texture with infiltrated papules and thickened plaques. Therapeutically, CAD is a problematic condition. Daily application of sun protection has to be combined with local or systemic immunosuppression. However, various therapeutic approaches including systemic corticosteroids and other systemic immunosuppressive agents are limited by severe side effects. Tacrolimus, a Streptomyces-derived immunosuppressive macrolide antibiotic and inhibitor of calcineurin, has been proven successful in various inflammatory skin diseases including atopic eczema, allergic contact dermatitis and photodermatoses without undesired side effects. We present a case of severe recalcitrant and nodular chronic actinic dermatitis responding to topically applied tacrolimus (Protopic). To avoid further relapses we continued this therapy twice a day over a period of 2 years. PMID- 16707889 TI - Sodium thiosulphate as a promising therapeutic option to treat calciphylaxis. AB - A 35-year-old haemodialysis-dependent woman with chronic renal failure developed large, very painful necrotic ulcers and necrosis on the thighs, buttocks and the abdomen with signs of fast progression. The skin biopsy specimens showed a broad necrosis of the epidermis and thrombosed dermal vessels with focal calcium deposits within the wall. In addition, laboratory findings presented an increased product of serum calcium and phosphate concentrations. Thus, we diagnosed calciphylaxis on the basis of clinical, biochemical and histopathological criteria. We initiated a therapy in which our patient was treated with intravenous sodium thiosulphate 3 times weekly. Already after 2 weeks of treatment, no new lesions were detectable and there was a dramatic pain relief. In the following 4 weeks, a successive decline of the ulcers and the healing of individual tissue defects could be seen. Four months after the start of the therapy, the patient underwent successful renal transplantation. Thus, the intravenous therapy of calciphylaxis with sodium thiosulphate might be a new effective alternative in the treatment of this life-threatening disease. PMID- 16707892 TI - Reticular erythematous mucinosis occurring in a brother and sister. AB - Reticular erythematous mucinosis (REM) is a rare, primary cutaneous mucinosis clinically characterized by a persistent reticular erythema on the mid chest and mid-upper back, and histologically by a mononuclear cell infiltrate and deposits of mucin in the dermis. To our knowledge, the present report of REM occurring in a Caucasian man and his sister is the first reported case of familial REM. Since a host-specific immune response to unknown antigens may be involved in the pathogenesis of this entity, human leukocyte antigen typing was determined and compared to those reported in autoimmune diseases. PMID- 16707891 TI - A case of epidermolysis bullosa acquisita with autoantibody to anti-p200 pemphigoid antigen and exfoliative esophagitis. AB - BACKGROUND: Some cases of a subepidermal blistering disease associated with autoantibodies to more than two antigens have been reported. OBSERVATION: A 52 year-old Japanese woman had pruritic blisters on almost the whole body as well as erosive lesions in the oral cavity and esophagus. A histological finding was subepidermal bullae. Direct immunofluorescence (IF) revealed a linear deposition of IgG, IgM and C3 at the epidermal basement membrane zone (BMZ). Indirect IF using human skin split by 1 M NaCl as a substrate showed IgG antibody reactive with the dermal side. By immunoblot analysis using normal human dermal extract, the 200-kDa and 290-kDa bands were detected. Indirect IF did not show any anti BMZ antibody activity, when using the skin of the patient with recessive dystrophic epidermolysis bullosa as a substrate. CONCLUSION: We regarded our case as epidermolysis bullosa acquisita with autoantibody to anti-p200 pemphigoid antigen. This is the second case in the literature associated with autoantibodies to these two antigens. PMID- 16707894 TI - Colocalization of vitiligo and lichen planus on scrotal skin: a finding contrary to the actinic damage theory. PMID- 16707895 TI - Erythema ab igne as an unexpected computer side-effect. PMID- 16707893 TI - A case of ulceronecrotic Mucha-Habermann disease with pulmonary involvement. PMID- 16707896 TI - Contrasted skin capacitance imaging of seborrheic keratoses and melanocytic nevi. PMID- 16707897 TI - Fever and penis ulcer in an 87-year-old man. PMID- 16707898 TI - Continuous education for renal transplant recipients. PMID- 16707899 TI - Evaluation of histopathological examination and direct immunofluorescence in the diagnosis of cicatricial alopecia. PMID- 16707900 TI - Calcinosis cutis universalis with joint contractures complicating juvenile dermatomyositis. PMID- 16707901 TI - Tension-free vaginal tape versus tension-free vaginal tape obturator in women with stress urinary incontinence. AB - BACKGROUND: To assess the efficacy and complications of tension-free vaginal tape (TVT) versus tension-free vaginal tape obturator in women with urodynamic stress incontinence. METHODS: Prospective, randomized study. Initially, 91 patients were included in the study and 89 of them were available at 12 months follow-up. Forty six patients were subjected to classic TVT procedure and 43 to transobturator vaginal tape from inside to outside (TVT-O) operation. There was no significant difference between the groups for age, BMI, menopausal status and prolapse. No patients had cystocele greater than stage I. Subjective and objective cure and improvement rate, mean operative time, hospital stay and complications incidence were assessed. RESULTS: Mean operative time was significant shorter in the TVT-O group (17.4 +/- 6.9 min) compared to the TVT group (26.7 +/- 8.6 min). There was no significant difference in the duration of hospital stay between two groups. The objective cure rate for TVT group was 89%, the improvement rate was 6.5%, the failure rate was 4.3% and the subjective cure rate 73.9%. The objective cure rate for TVT-O group was 90%, the improvement rate was 7.6%, the failure rate was 2.5% and the subjective cure rate 76.7%. The hemoglobin loss ranged between 1.0 +/- 0.5 g/dl for TVT group and 0.9 +/- 0.4 g/dl for TVT-O group. CONCLUSION: The TVT O technique presents success rates comparable to the classic TVT method in the short term. PMID- 16707902 TI - Increase in linkage information by stratification of pedigree data into gold standard and standard diagnoses: application to the NIMH Alzheimer Disease Genetics Initiative Dataset. AB - Patients diagnosed with a standard clinical method (subject to misclassification error) are often combined with patients diagnosed with a gold-standard method (with zero or very small misclassification error) in family-based studies of complex disease. For example, non-autopsied patients (NAP) are often included along with autopsy-proven (AP) patients in family-based studies of complex diseases, such as Alzheimer's disease (AD). Theoretical and simulation studies suggest that certain misclassification errors can result in severe reduction of power in genetic linkage and association analyses and that phenotype (or diagnostic) error can produce misleading results. Morton's test for heterogeneity can identify genomic regions where error may have led to loss in power. We applied this test to pedigree data from the NIMH Alzheimer's Disease Genetics Initiative Database separated into AP and NAP pedigrees. Morton's test identified one highly significant region of heterogeneity on chromosome 2. The source of the heterogeneity was due to significant indication of linkage in the AP pedigrees at position 109 cM (p value = 6.68 x 10(-5)) with no indication in the NAP pedigrees. Furthermore, Morton's test showed no evidence for heterogeneity on chromosome 19 in early-onset pedigrees that showed highly significant evidence for linkage in other published reports. These results suggest that supplementing linkage analysis with Morton's test can be usefully applied to genetic data sets that have AP and NAP samples, or other sample mixtures that include a 'gold standard' subgroup with reduced error rate, to increase power to detect linkage in the presence of diagnostic misclassification. PMID- 16707903 TI - Bone mass increase in puberty: what makes it happen? AB - It is now thought that the critical property of bone is strength rather than weight, and that control of bone strength is mainly exercised through the effect of the mechanical loads brought to bear on bone. Muscle contraction places the greatest physiological load on bone, and so the strength of bone must be adapted to muscle strength (the functional muscle-bone unit). The Utah paradigm of skeletal physiology [J Hum Biol 1998;10:599-605] provides a model of bone development that describes how bone structure is regulated by local mechanical effects that can be adjusted by the effects of hormones. The DONALD (Dortmund Nutritional and Anthropometric Longitudinally Designed) study analysed the interaction between the muscle and bone systems in males and females before and during puberty. This study found that differences between the genders in bone adaptation during puberty are at least partly driven by the influence of oestrogen in females. Testosterone seems to have no direct relevant effect on bone during puberty, but may be implicated in the greater amount of muscle mass achieved in boys compared with girls. PMID- 16707904 TI - Measuring bone mass in children: can we really do it? AB - Bone densitometry is used to assess skeletal health in clinical and research settings, with the goal of achieving reproducible measurements of bone mass that help to identify individuals predisposed to fracture. The search is now on for better methods of capturing additional factors that contribute to bone strength, including bone size, geometry, microarchitecture, and turnover rates. This has proved particularly challenging in growing children, whose bones continually change in size, shape, and mass. Dual energy X-ray absorptiometry is the preferred method for measuring bone mass in children, but the technique has several limitations, and interpreting the findings can be problematic. Peripheral quantitative computed tomography is a promising method for assessing bone mass and other indices correlating with bone strength, but a lack of precision and paediatric norms currently restricts its clinical utility. Although bone mineral density is predictive of future fracture risk in adults, the evidence in children is less conclusive, and a diagnosis of osteoporosis in a child should not be made on densitometric findings alone. Developing a clearer understanding of how measures of bone mass and strength correlate with bone fracture in children will help target preventive strategies for those in greatest need. PMID- 16707905 TI - Adolescents with childhood-onset GHD: how do we get them to peak bone mass? AB - The development of osteoporosis, with its attendant risk of fragility fracture, is in part related to the peak bone mass (PBM) achieved in early adulthood. Adolescence is a critical time for the acquisition of bone mass, with around 40% of skeletal mass being accrued during pubertal maturation. Growth hormone (GH) plays an integral role in the achievement of PBM after completion of linear growth, and several recent studies have suggested that GH replacement should continue in individuals with childhood-onset GHD until PBM has been attained - irrespective of the height achieved. In those with severe GHD after growth and pubertal development are complete, a seamless transition of GH therapy into adult life may be preferable to allowing a gap in GH treatment. The 'window of opportunity' concept for achieving PBM will, nevertheless, continue to be challenged by GHD teenagers who may resent the seamless continuation of GH replacement beyond adolescence. Preparation for this possibility should therefore begin during childhood, with all GHD teenagers being encouraged to remain on GH therapy until at least their mid-20s. PMID- 16707906 TI - Risk factors for contrast-induced nephropathy. AB - An increasing number of diagnostic imaging and interventional procedures require the use of radiographic contrast agents which has led to a parallel increase in the incidence of contrast-induced nephropathy (CIN). CIN is a serious clinical problem associated with increased morbidity and mortality, particularly in patients with chronic renal failure (see the Case Report). A key step to minimize CIN is to identify patients at risk of CIN. The aim of the present review was to summarize the knowledge about the risk factors of CIN, including the review of ultimate clinical research and developments. PMID- 16707907 TI - Education and dementia: a meta-analytic study. AB - Considerable controversy exists about the role of education in the risk of dementia. Individual studies have not been conclusive so far. To examine the hypothesis that lower education is associated with a higher risk of dementia, we carried out a meta-analysis. Observational studies published as of October 2005 that examined the association between education and risk of dementia were systematically reviewed. Relative risks (RRs) and odds ratios were extracted from cohort and case-control studies. We first compared the risk of dementia in subjects with high level of education with the risk of dementia in those with low educational level. In a subsequent analysis, we compared the risk of persons with high education with the risk of subjects with education level other than high (medium, low). We weighted log RRs for cohort studies or odds ratios by the inverse of their variances. Nineteen studies were included in our meta-analysis (13 cohort and 6 case-control studies). RRs for low versus high education level were: Alzheimer's disease (AD) 1.80 (95% CI: 1.43-2.27); non-AD dementias, 1.32 (95% CI: 0.92-1.88), and all dementias 1.59 (95% CI: 1.26-2.01). For low and medium versus high education level, the RRs were: AD 1.44 (95% CI: 1.24-1.67); non-AD 1.23 (95% CI: 0.94-1.61), and all dementias 1.33 (95% CI: 1.15-1.54). These results confirm that low education may be a risk factor for dementia, especially for AD. PMID- 16707908 TI - Clinical course of progressive multiple sclerosis in Brazilian patients. AB - OBJECTIVE: To describe the clinical course and outcome of multiple sclerosis with progressive onset in Brazilian patients. A total of 238 medical records were reviewed, 26 cases (10.9%) fulfilled Thompson criteria (2000), and 5.80% classified as primary progressive and 5.04% relapsing progressive according to Lublin and Reingold. STUDY POPULATION: 19 Caucasians and 7 non-Caucasians; male:female ratio 1.2:1, mean age at onset was 34 +/- 7.9 years. RESULTS: Non Caucasian patients had earlier onset of disease. The most common manifestations at onset were pyramidal and cerebellar (89% and 34.6%). After 11.3 +/- 6.35 years of disease more than 50% of the patients had involvement of most of their functional systems. No statistically significant differences were observed between the subgroups. CONCLUSION: The clinical course and outcome of progressive multiple sclerosis in Brazil, a tropical country with low prevalence, were very similar to those in the multiple sclerosis high prevalence areas. PMID- 16707909 TI - The bioactivity of plasma factors in focal segmental glomerulosclerosis. AB - Focal segmental glomerulosclerosis (FSGS) is a devastating form of nephrotic syndrome, often leading to end-stage renal failure after the failure of a succession of highly toxic therapies. It has long been thought to be caused by a circulating factor(s) that may be produced by cells of the immune system. Much research has focused on identifying such factor(s), including the development of a promising in vitro assay, which estimates glomerular permeability based on the swelling of isolated glomeruli in response to patients' plasma. This assay has also been used as the basis of testing plasma fractions for permeability activity, with no specific factor yet identified. Other studies have attempted to replicate proteinuria in whole animals, by injecting plasma or plasma fractions from focal segmental glomerulosclerosis patients, with inconsistent results. More recently there has been evidence that there may be either inhibitory or missing factor(s) in plasma, with respect to permeability. An additional major biological advance is a growing appreciation of the podocyte as the target cell in this disease, and an understanding of the key molecules involved. Putting together this knowledge, with the latest technological advances in protein identification, provides promising avenues towards finally solving the basis of this enigmatic disease. PMID- 16707910 TI - Efficacy of darbepoetin in doxorubicin-induced cardiorenal injury in rats. AB - This study was intended to elucidate the efficacy of an erythropoietin analog in cardiorenal dysfunction syndrome using a rodent model. Cardiorenal dysfunction was induced using doxorubicin hydrochloride (DXR). Lower doses (3 microg/kg) and higher doses (30 microg/kg) of darbepoetin alfa (DA) were used for intervention. Blood examinations for creatinine, blood urea nitrogen, iron, and hemoglobin were performed until 11 weeks after starting DA administration. Urine collection was performed 10 weeks after starting DA, and protein, iron, and N-acetyl-beta-D glucosaminidase levels and antioxidation capacity of DA were determined. The dry left ventricular heart weight was measured, when the animals were sacrificed 11 weeks after starting DA administration. Histological analyses were performed for interstitial fibrotic changes and iron deposition in the kidney. Administration of DA markedly improved anemia to the normal control level and significantly alleviated DXR-induced increases of creatinine, blood urea nitrogen, renal interstitial fibrosis, renal iron deposition, and dry left ventricular weight, but serum and urinary iron and urinary protein and N-acetyl-beta-D glucosaminidase levels were unchanged. The urinary total radical-trapping antioxidant capacity was improved to the normal control level in DA-treated animals. DA reduced the DXR-induced cardiorenal injury. This improvement was achieved, when anemia was corrected to the normal control level. PMID- 16707911 TI - Effects of growth hormone substitution therapy on cognitive functioning in growth hormone deficient patients: a functional MRI study. AB - Patients with childhood-onset growth hormone (GH) deficiency (GHD) show impairments in mood and cognitive functioning which may resolve following GH substitution. Brain functional magnetic resonance imaging (fMRI) during performance of a memory task was used to assess the cerebral activity of such patients. Thirteen childhood-onset GHD patients (mean age 27.3 +/- 6.9 years) were included in a double-blind, placebo-controlled study. The effects of 6 months of GH replacement or placebo therapy were studied using neuropsychological tests and fMRI. One patient was excluded from the study due to noncompliance with the protocol. Six months of GH substitution in these GHD patients resulted in improved memory functioning, both for long-term and working memory. fMRI showed activations during the working memory task in prefrontal, parietal, motor, and occipital cortices, as well as in the right thalamus and anterior cingulate cortex. Decreased activation in the ventrolateral prefrontal cortex was observed after GH treatment as compared with placebo treatment, indicating decreased effort and more efficient recruitment of the neural system involved. It can be concluded that GH treatment for 6 months improved the long-term as well as the working memory in patients with GHD, and this was associated with decreased brain activation in the ventrolateral prefrontal cortex. GH substitution in GHD patients is beneficial for cognitive functioning, the effects of which can be visualized by means of neuroimaging. PMID- 16707912 TI - Effect of insulin on the brain activity obtained during visual and memory tasks in healthy human subjects. AB - Insulin receptors are found throughout the brain, particularly in the hippocampus, although the impact of insulin on memory is unclear. The purpose of this study was to examine the effect of insulin on event-related potentials in response to a standard memory task and visual evoked potentials (VEPs) during exposure to a reversing checkerboard. We hypothesized that insulin would decrease P300 magnitude and latency during the presentation of previously observed stimuli, but would have no effect on VEPs. Sixteen humans participated in two euglycemic clamp studies with somatostatin performed in random order in which serum insulin levels were either suppressed (14 +/- 1 pmol/l) or increased by insulin infusion (433 +/- 40 pmol/l). At steady state, event-related potentials and then VEPs were collected using a 32-electrode cap. The major finding was that the P300 amplitude measured during the identification of an object as old was significantly smaller over parietal regions when insulin was infused than when no insulin was provided. Insulin was without effect on the VEPs. We conclude that insulin has region- and task-specific effects on neuronal activation. While the P300 amplitude measured during the presentation of an old object was reduced during insulin infusion, the hormone was without effect on VEPs. PMID- 16707913 TI - Characteristics of post-stroke depression in Japanese patients. AB - Post-stroke depression (PSD) has an important impact on the quality of life of patients with stroke. We studied 100 stroke patients (mean age+/-SD: 64.6+/-11.6, range: 32-85 years) in the subacute phase (2-5 weeks after onset) and investigated the prevalence and clinical correlates of PSD in the subacute phase. The prevalences of PSD and major depression in the subacute phase evaluated by the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, were 20.0 and 5.0%, respectively. These values were lower than those reported in Caucasian studies. The Zung Self-Rating Depression Scale (SDS) did not underestimate the presence of PSD and was thus considered valuable for the initial screening of PSD. However, SDS may overestimate PSD especially in patients with anxiety disorders or somatoform disorders and vice versa. Apathy was observed in 40.2% of patients; however, it was widely distributed and observed even in patients without serious depression. PMID- 16707914 TI - BDNF Val66Met polymorphism is associated with body mass index in healthy adults. AB - Although recent studies suggest a possible relationship between the brain-derived neurotrophic factor Val66Met polymorphism and eating disorders, no study has examined the possibility that the Met-Met genotype is associated with a lower body mass index (BMI) in healthy individuals. We examined this possibility in 481 adults (age range 18-82 years) without significant medical or psychiatric history. After adjusting for gender, analysis of covariance showed that persons with the Met-Met genotype had a lower BMI than those with the Val-Met/Val-Val genotypes (22.28 +/-3.77 vs. 24.72+/-4.81). A similar, though nonsignificant, trend emerged when comparing all three genotypes separately. These findings suggest a possible relationship between Val66Met polymorphism and BMI in healthy adults. Further work is needed to clarify possible mechanisms for this relationship. PMID- 16707916 TI - Opinions on cochlear implant use in senior MED-EL patients. AB - This study assessed the subjective benefits of cochlear implants in senior patients wearing a MED-EL device. Data was compared with previous studies to assess the influence of recent speech-coding strategies and behind-the-ear speech processors; users of the behind-the-ear device and the body-worn device were also compared. An adaptation of the Nucleus 22-channel survey was sent to 141 cochlear implant users. The survey assesses perception of the device, communication benefits, handling the device and quality of life. Ninety-five surveys were returned. Results demonstrate that advanced technology provides greater benefit now than 9-13 years ago, notably: improved listening across noise, better understanding on the telephone and better speech perception. Users of the behind the-ear device did not report more difficulties than body-worn device users but demonstrated better performance. Results show a positive outcome for cochlear implantation in a MED-EL seniors group. PMID- 16707915 TI - Positron emission tomography imaging of risperidone augmentation in serotonin reuptake inhibitor-refractory patients. AB - We studied 15 nondepressed patients with obsessive-compulsive disorder (OCD) who were nonresponders to serotonin reuptake inhibitors with an additive trial of risperidone. Positron emission tomography with (18)F-deoxyglucose and magnetic resonance imaging was obtained at baseline and following 8 weeks of either risperidone or placebo in a double-blind parallel group design. Risperidone treatment was associated with significant increases in relative metabolic rate in the striatum, cingulate gyrus, the prefrontal cortex, especially in the orbital region, and the thalamus. Four of 9 patients who received risperidone showed clinical improvement (CGI score of 1 or 2 at 8 weeks) while none of the 6 patients who received placebo showed improvement. Patients with low relative metabolic rates in the striatum and high relative metabolic rates in the anterior cingulate gyrus were more likely to show a clinical response. These metabolic predictors of clinical response are consistent with earlier PET studies showing similar prediction when either neuroleptics or serotonin reuptake inhibitor treatments are administered individually. Our results are consistent with a frontostriatal circuit change related to both dopaminergic and serotonergic systems and with the presence of psychopharmacological subtypes within OCD. PMID- 16707917 TI - Lipoma of the deep lobe of the parotid gland: report of 3 cases. AB - Since lipomas are common soft-tissue neoplasms in other regions of the body, they are found only rarely in the parotid gland and are, therefore, often not considered in the initial differential diagnosis of parotid gland tumours. Even rarer are lipomas arising in the deep lobe of the parotid gland with only 6 cases presented in the literature. Lipomas originating in the parapharyngeal space are also extremely rare. When there is any suspicion, computed tomography and especially magnetic resonance imaging are of high diagnostic value. Fine-needle aspiration cytology seems not to contribute much to an accurate diagnosis. None of these preoperative examinations allows an absolute reliable distinction between a lipoma and a liposarcoma. The preferred treatment is complete surgical excision which will minimize the possibility of a recurrence and will also lead to a definitive diagnosis. We report 3 patients with a lipoma of the deep lobe of the parotid gland, extending into the parapharyngeal space, and discuss differential diagnosis and radiological features of lipomas in this rare location. PMID- 16707918 TI - Snake C-type lectin-like proteins and platelet receptors. AB - Snake venoms are complex mixtures of biologically active proteins and peptides. Many affect haemostasis by activating or inhibiting coagulant factors or platelets, or by disrupting endothelium. Snake venom components are classified into various families, such as serine proteases, metalloproteinases, C-type lectin-like proteins, disintegrins and phospholipases. Snake venom C-type lectin like proteins have a typical fold resembling that in classic C-type lectins such as the selectins and mannose-binding proteins. Many snake venom C-type lectin like proteins have now been characterized, as heterodimeric structures with alpha and beta subunits that often form large molecules by multimerization. They activate platelets by binding to VWF or specific receptors such as GPIb, alpha2beta1 and GPVI. Simple heterodimeric GPIb-binding molecules mainly inhibit platelet functions, whereas multimeric ones activate platelets. A series of tetrameric snake venom C-type lectin-like proteins activates platelets by binding to GPVI while another series affects platelet function via integrin alpha2beta1. Some act by inducing VWF to bind to GPIb. Many structures of these proteins, often complexed with their ligands, have been determined. Structure-activity studies show that these proteins are quite complex despite similar backbone folding. Snake C-type lectin-like proteins often interact with more than one platelet receptor and have complex mechanisms of action. PMID- 16707920 TI - Bothrojaracin, a Bothrops jararaca snake venom-derived (pro)thrombin inhibitor, as an anti-thrombotic molecule. AB - Bothrojaracin (BJC) is a selective and potent thrombin inhibitor (KD = 0.6 nM) which also binds to prothrombin on proexosite I (KD = 175 nM). Incubation of BJC with human or rat plasma produced a band that co-migrates with purified prothrombin-BJC complex. We further analyzed the in vivo anti-thrombotic effect of BJC on a venous thrombosis model in rats that combines stasis and hypercoagulability. The administration of 1 mg/kg (i.v.) doses of BJC decreased thrombus weight by approximately 95%. Evaluation of the in vivo effect of BJC in mice using a pulmonary thromboembolism model induced by thrombin showed that BJC protects 100% of mice from death. Altogether, our data show that BJC is a potent anti-thrombotic agent that could further help the development of new prothrombin directed drugs. PMID- 16707919 TI - Structure-function relationships of C-type lectin-related proteins. AB - The structural and functional studies of the first identified C-type lectin-like protein (CLP), blood coagulation factor IX/factor X-binding protein (IX/X-bp), have been instrumental in defining how new functionally heterodimeric CLPs are generated from monomeric carbohydrate recognition domain in C-type lectins by three-dimensional domain swapping. The crystal structures of gamma carboxyglutamic acid domains of coagulation factors X and IX have recently been clarified in structural studies of complexes between the gamma-carboxyglutamic acid domain of factors X and X-bp (a venom CLP) and between the gamma carboxyglutamic acid domain of factors IX and IX-bp (a venom CLP). PMID- 16707921 TI - Scratching below the surface: wound healing and alanine mutagenesis provide unique insights into interactions between eristostatin, platelets and melanoma cells. AB - To study the molecular mechanism of the disintegrin eristostatin, cellular functional studies were performed using ten recombinant alanine mutants. ADP induced platelet aggregation revealed critical contributions of seven residues within the 'RGD loop' (R24, R27, G28, N31) and C-terminus (W47, N48, G49) of this disintegrin. Using an in vitro scratch wound healing assay, four human melanoma cell lines yielded similar results when exposed to wildtype eristostatin. All eristostatin-treated cells healed less of the wounded area than control conditions. This phenomenon was reproduced when using fibronectin as the matrix. C8161 cells showed significant delay in wound closure with the N-terminal mutant P4A but not with R24A or G28A. Evidence from our laboratory and others suggests neither alpha IIb, alpha 4 nor alpha 5 integrins are directly involved in eristostatin's interactions. Eristostatin did not affect the number of melanoma cells in culture after 24 h or the development of apoptosis. However, phosphorylation studies performed after these melanoma cells were exposed to eristostatin revealed changes in several tyrosine phosphorylated molecules. PMID- 16707922 TI - Contortrostatin, a snake venom disintegrin with anti-angiogenic and anti-tumor activity. AB - Disintegrins are soluble peptides found in snake venom. They bind to Arg-Gly-Asp (RGD)-responsive integrins with high affinity (nM range) and block integrin function. Contortrostatin (CN), the disintegrin from southern copperhead venom, is a homodimer with a molecular weight of 13,500. Each chain contains 65 amino acids with an Arg-Gly-Asp motif. CN has anti-invasive and anti-adhesive activity on tumor cells and endothelial cells in vitro, and binds to integrins alphavbeta3, alphavbeta5, and/or alpha5beta1. In vivo studies using the human metastatic breast cancer cell line MDA-MB-435, in an orthotopic xenograft model in nude mice, revealed that CN has potent anti-tumor and anti-angiogenic activity. Recent studies have employed an intravenous liposomal delivery procedure. Liposomal delivery of CN has also been shown to provide effective in vivo anti-tumor and anti-angiogenic activity in a human ovarian cancer animal model. PMID- 16707923 TI - Development of a novel recombinant disintegrin, contortrostatin, as an effective anti-tumor and anti-angiogenic agent. AB - Contortrostatin (CN) (Mr 13,500 Da) is a novel homodimeric disintegrin isolated from the venom of Agkistrodon contortrix contortrix (Southern Copperhead) snake and displays two RGD motifs (one on each chain), which modulate its interaction with integrins on tumor cells and angiogenic vascular endothelial cells. In previous studies, we have shown that native CN administered in a liposomal formulation exhibits potent anti-angiogenic and tumor growth inhibitory activities. Current isolation of the protein from crude venom is difficult and prohibitively expensive for translation into the clinic. In this report, we describe a method amenable to large-scale production of a soluble monomeric form of recombinant CN with biologic activity; the protein is expressed directly in the cytoplasm of an engineered bacterial system with an expression yield of approximately 20 mg/l of culture. We present here the in vitro assays as well as the anti-tumor and anti-angiogenic evaluation of liposomal recombinant CN in an orthotopic, xenograft model of human breast cancer. PMID- 16707924 TI - Hemextin AB complex--a snake venom anticoagulant protein complex that inhibits factor VIIa activity. AB - Snake venom is a veritable gold mine of bioactive molecules, capable of binding to a wide variety of pharmacological targets, including the blood coagulation cascade. Here, we report the isolation and characterization of two synergistically acting anticoagulant three-finger proteins, hemextin A and hemextin B, from the venom of Hemachatus haemachatus (African Ringhals cobra). Hemextin A but not hemextin B exhibits mild anticoagulant activity. However, hemextin B interacts with hemextin A and forms a complex (hemextin AB complex), and synergistically enhances its anticoagulant potency. Prothrombin time assay showed that these two proteins form a 1:1 complex. Using a 'a dissection approach', we found that hemextins A and AB complex prolong clotting by inhibiting extrinsic tenase activity. Further studies showed that hemextin AB complex potently inhibits the proteolytic activity of factor VIIa (FVIIa) and its complexes. Kinetic studies showed that hemextin AB complex is a non-competitive inhibitor of FVIIa-soluble tissue factor proteolytic activity with a K(i) of 25 nM. Hemextin AB complex is the first reported natural inhibitor of FVIIa that does not require either tissue factor or factor Xa scaffold to mediate its inhibitory activity. Molecular interactions of hemextin AB complex with FVIIa/tissue factor-FVIIa may provide a new paradigm in the search for anticoagulants inhibiting the initiation of blood coagulation. PMID- 16707925 TI - Comparison of textilinin-1 with aprotinin as serine protease inhibitors and as antifibrinolytic agents. AB - Textilinin-1 (Q8008) was isolated from the venom of the Pseudonaja textilis and has a 47% sequence identity to the antihaemorrhagic therapeutic agent aprotinin. When equimolar concentrations of enzyme and aprotinin were pre-incubated, plasmin was inhibited 100%, plasma kallikrein 58%, and tissue kallikrein 99%. Under the same conditions, textilinin-1 inhibited plasmin 98%, plasma kallikrein 16% and tissue kallikrein 17%. Whole blood clot lysis was inhibited strongly by both aprotinin and textilinin-1, as shown by thrombelastography. At 2 microM inhibitor lysis initiated by t-PA was greater than 99% inhibited by aprotinin (LY60 = 0.4 +/- 0.1) whereas textilinin-1, inhibited lysis by 91% (LY60 = 8.9 +/- 0.7). The same trend was found with the lysis of euglobulin fractions. From these data textilinin-1 appears to be a more specific plasmin inhibitor than aprotinin but aprotinin inhibits clot lysis to a greater extent. PMID- 16707926 TI - Identification and localization of heparin-binding region of snake venom VEGF and its blocking of VEGF-A165. AB - Vascular endothelial growth factor (VEGF-A165) displays diverse effects through binding to its receptor, KDR (kinase domain-containing receptor). Heparan sulfate/heparin-like molecules greatly contribute to their interaction. Indeed, the mitogenic potency of VEGF-A165 lacking the C-terminal heparin-binding region is less than 1% compared with intact VEGF-A165. We previously found novel heparin binding VEGFs, designated VEGF-F that specifically recognizes KDR in snake venoms. VEGF-Fs almost completely lack the C-terminal heparin-binding region compared with VEGF-A165, despite their heparin-binding potential. In this study, we attempted to identify the heparin-binding region of VEGF-F using synthetic peptides. We have demonstrated that the heparin-binding site of VEGF-F is located in its C-terminal region, particularly localized on the N-terminal portion of this region. Furthermore, a synthetic peptide of this region blocks the biological activity of VEGF-A165 in vitro and in vivo. PMID- 16707927 TI - C-terminal heparin-binding peptide of snake venom VEGF specifically blocks VEGF stimulated endothelial cell proliferation. AB - Vascular endothelial growth factor-A165 (VEGF-A165) exhibits diverse biological effects through binding to its receptor KDR (VEGFR-2). Heparin-like molecules are known to modulate their interaction. There have been reports that VEGF-A lacking the C-terminal heparin binding region significantly reduced mitogenic activity. Recently, we found novel heparin-binding VEGFs from snake venoms, designated VEGF Fs, which specifically recognize kinase domain containing receptor (KDR). The C terminal heparin-binding region is almost completely absent in VEGF-Fs when compared with other heparin-binding VEGFs, despite their heparin-binding potential. In this congress, we report that the C-terminal heparin-binding region of VEGF-F specifically/preferentially interacts with the VEGF-bondable heparin/heparan sulfate, but not with those associated with bFGF or TFPI. We also present the identification of a VEGF receptor-binding protein from the venom of eastern cottonmouth (Agkistrodon piscivorus piscivorus). Sequence analysis revealed the isolated KDR-binding protein (designated KDR-bp) is identical to Lys49PLA2, an inactive PLA2 homologue with strong myoxicity. KDR-bp binds to the extracellular domain of KDR with subnanomolar affinity. The interaction between KDR-bp and KDR was blocked by VEGF-A165, and KDR-bp specifically inhibited VEGF A165-stimulated endothelial cell proliferation, indicating KDR-bp is an antagonistic ligand for KDR. This is the first observation demonstrating that an exogenous factor antagonizes the VEGF receptor, furthermore, it is the first identification of the target molecule of the myotoxic PLA2 from viper venom. PMID- 16707928 TI - Serine proteases affecting blood coagulation and fibrinolysis from snake venoms. AB - Snake venom proteases, in addition to their contribution to the digestion of the prey, affect various physiological functions. They affect platelet aggregation, blood coagulation, fibrinolysis, complement system, blood pressure and nervous system. This review provides a ready reference for serine proteases that interfere in blood coagulation and fibrinolysis. They exhibit their activity by activation of specific zymogens of coagulation factors. These serine proteases serve as tools to study molecular details in the activation of specific factors involved in coagulation and fibrinolytic cascades and are useful in treating various thrombotic and hemostatic conditions. PMID- 16707929 TI - Gene structures of trocarin D and coagulation factor X, two functionally diverse prothrombin activators from Australian rough scaled snake. AB - Activation of prothrombin to thrombin is the key reaction in blood coagulation cascade. We have recently shown that Australian rough scaled snake, Tropidechis carinatus, possesses two parallel prothrombin activator systems. Trocarin D, a venom prothrombin activator produced in the venom gland, plays an offensive role as a toxin, whereas factor X is produced in the liver and plays a role in the hemostatic mechanism. These two proteins are structurally similar and have identical domain architecture. Because of the differences in their physiological roles, and tissue-specific expression, we determined the gene structure of these closely related proteins. Both the genes have eight exons similar to all mammalian factor X genes. All the exon-intron boundaries of these two genes are at the same position and the splice junctions are almost identical. Partial sequencing of the introns shows that they share a very high degree of sequence identity indicating that the gene duplication is a recent event. Further studies on the characterization of these two genes particularly the promoter regions are in progress. PMID- 16707930 TI - Vipera lebetina venom contains all types of snake venom metalloproteases. AB - Snake venoms contain four classes of metalloproteases that all have a typical zinc-chelating sequence (HEXXHGXXH). N-terminal sequences and internal sequences of different purified metalloproteases were determined using Edman sequencing and LC MS/MS technique. Oligonucleotides were designed and used as primers for cDNA cloning from Vipera lebetina venom gland cDNA library. We found that isoforms of fibrinolytic enzyme lebetase Le-4 and Le-3 are synthesized in different way: Le-4 is synthesized as P-I type metalloprotease, Le-3 is synthesized with disintegrin like domain as P-II type protease and processed post-translationally. An endothelial cell apoptosis-inducing heterodimeric glycosylated metalloprotease, V. lebetina apoptosis-inducing protease (VLAIP), belongs to P-III type containing metalloprotease, disintegrin-like and cysteine-rich domains. All these enzymes hydrolyze the Aalpha-chain and more slowly the Bbeta-chain of fibrinogen. Treatment of HUVEC cells with VLAIP induces changes in the attachment of cells to the substrate and causes apoptosis. V. lebetina venom contains also P-IV type specific coagulant factor X activator (VLFXA) that cleaves the Arg52-Ile53 bond in the heavy chain of human factor X. VLFXA is a glycoprotein composed of a heavy chain and two C-type lectin-like light chains linked by disulfide bonds. The heavy and light chains of VLFXA are synthesized from different genes. PMID- 16707931 TI - Non-clinical and clinical characterization of a novel acting thrombolytic: alfimeprase. AB - Alfimeprase (ALF) is a recombinant, truncated form of fibrolase, a directly fibrinolytic zinc metalloproteinase that was first isolated from the venom of the Southern copperhead snake (Agkistrodon contortrix contortrix). ALF has direct proteolytic activity against the fibrin(ogen) Aalpha chain. ALF can be covalently bound and neutralized by serum alpha2-macroglobulin, a prevalent mammalian protease inhibitor. Preclinical pharmacology studies have shown that thrombolysis with ALF is up to 6-times more rapid than with select plasminogen activators. Additional studies suggest that intra-thrombus ALF has the potential to be a fast and effective thrombolytic without generation of a systemic lytic state. Investigations of phases 1 and 2 indicate that ALF is active and generally well tolerated. This paper reviews the biochemical characteristics of ALF and a review of the preliminary clinical experience in subjects with acute peripheral arterial occlusion and in those with central venous access device occlusion. PMID- 16707932 TI - Antihemostatic molecules from saliva of blood-feeding arthropods. AB - The ability to feed on vertebrate blood has evolved many times in various arthropod clades. Each time this trait evolves, novel solutions to the problem posed by vertebrate hemostasis are generated. Consequently, saliva of blood feeding arthropods has proven to be a rich source of antihemostatic molecules. Vasodilators include nitrophorins (nitric oxide storage and transport heme proteins), a variety of peptides that mimic endogenous vasodilatory neuropeptides, and proteins that catabolize or sequester endogenous vasoconstrictors. A variety of platelet aggregation inhibitors antagonize platelet responses to wound-generated signals, including ADP, thrombin, and collagen. Anticoagulants disrupt elements of both the intrinsic and extrinsic pathways. Molecular approaches (termed 'sialomics') to characterize the full inventory of mRNAs transcribed in salivary glands have revealed a surprising level of complexity within a single species. Multiple salivary proteins may be directed against each component of hemostasis, resulting in both redundancy and in some cases cooperative interactions between antihemostatic proteins, as in the case of the Rhodnius prolixus apyrase (which hydrolyzes ADP) and Rhodnius platelet aggregation inhibitor 1 (which sequesters ADP). The complexity and redundancy of saliva ensures an efficient blood meal for the arthropod, but it also provides a diverse array of novel antihemostatic molecules for the pharmacologist. PMID- 16707933 TI - Exploring new molecules and activities from Lonomia obliqua caterpillars. AB - Skin contact with Lonomia caterpillar bristles causes a consumptive coagulopathy. From a cDNA library we cloned and expressed a prothrombin activator (rLopap) in active form, and from the bristles extract we characterized a FX activator (Losac). Several clones were sequenced and analyzed by expressed sequence tags. A database of about 1,270 sequences was constructed and deposited in NCBI (CX815710 CX817210) [corrected] Both the native protein from the venom (Lopap) and the recombinant form (r-Lopap) promoted prothrombin hydrolysis, generating prethrombin-2, F1.2 and thrombin. Losac is a single-chain (43 kDa) protein that cleaves the FX heavy chain producing FXaalpha. In HUVECs rLopap and Losac are able to modulate cell survival by preventing apoptosis. rLopap increases NO and PGI2 concentration and Losac induces t-PA expression. Finally, to identify the venom proteins related to human envenomation, a 2D electrophoresis map is being performed as an attempt to find the major toxins recognized by the anti-lonomia venom. PMID- 16707934 TI - The use of snake venom-derived compounds for new functional diagnostic test kits in the field of haemostasis. AB - Pentapharm Ltd. has a long tradition of producing and proceeding snake venom components as APIs and tools in pharmaceutical and diagnostic applications, beginning with the snake venom-derived product Reptilase about 60 years ago. In recent years diagnostic test kits in the field of haemostasis have been developed, like Pefakit APC-R Factor V Leiden and Pefakit PiCT, which make use of one or more snake venom compounds as key reactants. The venom-derived compounds are responsible for many outstanding properties of these tests and account for their top ranking performance qualities confirmed in non-clinical and clinical studies. Both tests are on the market in Europe. Pefakit APC-R Factor V Leiden is also marketed in the US since January 2005. Research on snake venom-derived compounds for diagnostic and pharmaceutical use is ongoing. PMID- 16707935 TI - The potential of Bothrops moojeni venom in the field of hemostasis. Established use and new insights. AB - Early studies in the 1930s on the venom of South American Lancehead snakesofthe Bothrops genuslead to the discovery of compounds active in blood coagulation such as batroxobin and botrocetin. The scope of our investigations is to have a deeper look at the crude venom of B. moojeni using state-of-the-art proteomics methods, as well as newly developed bioassays screening for activities in the different fields of application. The proteomics techniques used up to now have included different chromatography methods, mass spectrometry, and bio-computing. The bioassays are focussed on enzymatic and other activities in the field of hemostasis and fibrinolysis. Besides the known activities several new and interesting ones have been found. They still need to be studied and confirmed in more specific supplementary assays. PMID- 16707938 TI - Best practices in teaching design: using online learning resources in MERLOT. PMID- 16707939 TI - Nursing-oriented Web resources. PMID- 16707940 TI - An interactive self-paced electrocardiography electrode placement tutorial. PMID- 16707941 TI - The International Council of Nurses innovations database: a Web-based resource designed to capture and share nursing innovations worldwide. PMID- 16707942 TI - Native American Elders Health Care Series. PMID- 16707943 TI - The use of personal digital assistants at the point of care in an undergraduate nursing program. AB - This study examined the relationships between the use of personal digital assistants and self-efficacy and the preparation for medication administration among second-year Bachelor of Science in Nursing students in a medical-surgical clinical environment. By using a controlled experimental method, the study attempted to support claims about the educational benefits of personal digital assistants which have generally been reported in more descriptive and anecdotal formats. The sample consisted of 36 students, of which two groups had personal digital assistants and two groups served as a control. The control groups were provided with paper resources equivalent to the software provided by the personal digital assistants. Findings showed a significant increase in self-efficacy in the groups with personal digital assistants. PMID- 16707944 TI - Developing and establishing online student learning communities. AB - Developing effective learning communities is an important component of Web-based courses. Learning communities offer a social context for learning that greatly enhances the knowledge acquisition of all involved parties. This article describes the development of an effective learning community among Web-based RN BSN students. The characteristics of the cohort leading to an effective learning community included supportiveness, open sharing of oneself, and socialization. PMID- 16707946 TI - A public health nursing informatics data-and-practice quality project. AB - Automation of nursing documentation in client records is one of today's formidable challenges in healthcare settings. Some stakeholders view standardized nursing language as a fundamental component of an automated record, with the end goal of generating valid, reliable data on nursing services and client outcomes. However, few successful applications of standardized nursing language for practice standards and data generation exist in the literature. A joint practice and-data quality project based on standardized nursing language in an automated platform was undertaken by four local public health departments. Outcomes of this project included articulation of standards for nursing practice and creation and implementation of a sound data quality infrastructure for nursing documentation of client assessments, service delivery information, and client outcomes. PMID- 16707947 TI - Computer courses in the undergraduate nursing curriculum in Turkey. AB - This study surveys computer courses in undergraduate nursing schools in Turkey. To accomplish this, the investigator gave an assignment to first-year students at the Ege University School of Nursing as part of a computer course in 2003-2004. The assignment consisted of having students use their computer skills and do research on the Internet to obtain information about computer classes at other Turkish nursing schools. The objective of this assignment was to correspond by e mail with first-year students at these institutions. Because of this assignment, at least 70 contacts were established, and information has been exchanged between nursing students at 45 different universities in Turkey. This study shows that one method used by undergraduate nursing schools in Turkey to educate their students in modern nursing practices is to integrate computer courses in their curricula, thereby providing students important practical technology skills. PMID- 16707948 TI - Falls in hospitalized patients: can nursing information systems data predict falls? AB - Falls among inpatients are the most frequently reported critical incidents in hospitals and can have tragic consequences that affect morbidity and mortality. The present study aimed to determine whether certain nursing units of care identified on patient care plans can be used to predict falls among hospitalized inpatients. A retrospective analysis of 7167 inpatient admissions in the 2002 calendar year was conducted. Faller status was ascertained from the hospital's accident and incident monitoring system, and nursing units of care activated in the hospital's nursing information system were identified. Twelve nursing units of care predicted falls. Logistic regression analyses showed that nursing units of care related to patient safety, confusion, incontinence, medication, mobility, and sleep were significant risk factors for falls among inpatients. The number of nursing units of care activated also predicted falls. Data collected from nursing information systems can be used to identify patients at high risk of falls. PMID- 16707954 TI - Inflammation markers in young post-myocardial patients exhibiting various expressions of classic coronary risk factors. AB - OBJECTIVE: We investigated inflammation markers in young post-myocardial infarction patients exhibiting various expressions of classical risk factors. METHODS: Forty-one male patients with high (n=20) and low (n=21) expression of classical risk factors (risk of coronary events calculated by the prospective cardiovascular Munster study program high or low, respectively), on average 44 years old, who were in the stable phase after myocardial infarction (on average 20.5 months after myocardial infarction) were included in the study. The control group consisted of 25 healthy, age-matched men. The following inflammation markers were measured: leukocyte count, high-sensitive C-reactive protein, interleukin-6, tumor necrosis factor-alpha, intracellular adhesion molecule-1, vascular cellular adhesion molecule-1, selectin-P and selectin-E. RESULTS: No differences in the levels of leukocytes, high-sensitive C-reactive protein, tumor necrosis factor-alpha, intracellular adhesion molecule-1, vascular cellular adhesion molecule-1, selectin-P and selectin-E were found between the group of patients and the controls. In contrast, interleukin-6 was significantly (P<0.01) elevated in the group of patients with high [2.5 (1.9-5.3) ng/ml] and low [3.2 (1.5-8.4) ng/ml] expression of risk factors compared with the controls [1.4 (0.9 2.3) ng/ml]. Significantly, there was no difference in interleukin-6 between the two groups of patients. CONCLUSIONS: We did not find differences in inflammation markers between young post-myocardial infarction patients with or without classical risk factors. Thus, it seems that the presence of (treated) risk factors or their absence does not affect the levels of inflammation markers in the stable period after myocardial infarction. Importantly, we found similarly elevated interleukin-6 in both groups of patients, most probably indicating slight local vascular inflammation. Interleukin-6 appears to be the most suitable marker of vascular inflammation in post-myocardial infarction patients who are aggressively treated pharmacologically. PMID- 16707955 TI - Carotid intima-media thickness in coronary slow flow: relationship with plasma homocysteine levels. AB - BACKGROUND AND OBJECTIVE: Coronary slow-flow phenomenon is characterized by delayed opacification of coronary vessels in a normal coronary angiogram. Although clinical and pathological features have been previously described, the underlying pathophysiology has not been fully elucidated. Thus, it still remains to be determined whether either microvascular or epicardial diffuse atherosclerotic disease is related to slow flow. In this study, we aimed to determine the carotid artery intima-media thickness, which is a marker of atherosclerosis in patients with coronary slow flow, and its possible relationship with the total homocysteine level. METHOD: The study population consisted of 88 patients who underwent coronary angiography because of typical and quasi-typical symptoms of angina. Forty-four patients with angiographically proven coronary slow flow and 44 individuals with normal coronary flow pattern with similar risk profiles were enrolled in the study. Coronary flow patterns of the latter were determined by the thrombolysis in myocardial infarction frame count method. Intima-media thickness was measured by recording ultrasonographic images of both the left and the right common carotid artery with a 12-MHz linear array transducer. Plasma homocysteine, folate and B12 levels were measured from blood samples. RESULTS: Plasma homocysteine levels (mumol/l) and carotid intima media thickness (mm) of patients with coronary slow flow were found to be significantly higher than that of controls (12.4+/-4.9 vs. 8.5+/-2.8, P=0.0001; 0.75+/-0.08 vs. 0.69+/-0.06, P=0.0001, respectively). The plasma folate level (ng/ml) was lower in coronary slow-flow patients than in controls (13.8+/-4.4 vs. 16.5+/-5.6, P=0.014). The plasma homocysteine level was significantly positively correlated with the mean thrombolysis in myocardial infarction frame count and intima-media thickness of the carotid artery in correlation analysis (r=0.58, P=0.0001; r=0.41, P=0.0001; respectively). CONCLUSION: Homocysteine levels and carotid intima-media thickness increased but folate levels decreased in patients with coronary slow flow. The present findings allow us to conclude that the possible disturbance in the metabolism of homocysteine in patients with coronary slow flow may have a role in the etiopathogenesis of this phenomenon by causing generalized atherosclerosis. PMID- 16707956 TI - Impact of metabolic syndrome on myocardial perfusion grade after primary percutaneous coronary intervention in patients with acute ST elevation myocardial infarction. AB - AIMS: Metabolic syndrome with its associated cardiovascular risk factors and prothrombotic, procoagulant and proinflammatory properties and its detrimental effects on coronary microcirculation may play a role in the occurrence of poor myocardial perfusion after primary percutaneous coronary intervention in patients with acute myocardial infarction. Accordingly, this study was designed to evaluate the association between metabolic syndrome and myocardial perfusion grade in patients with acute myocardial infarction undergoing primary percutaneous coronary intervention. MATERIAL AND METHODS: The study population included 283 consecutive patients (229 men, mean age=62+/-8 years) admitted to our hospital with ST-elevation myocardial infarction and who underwent primary percutaneous coronary intervention. Thrombolysis in myocardial infarction (TIMI) myocardial perfusion grade (TMPG) was graded densitometrically on the basis of visual assessment of relative contrast opacification of the myocardial territory subtended by the infarct vessel in relation to epicardial density. Metabolic syndrome was diagnosed according to the National Cholesterol Education Program Adult Treatment Panel III criteria. Patients were divided into two groups on the basis of the myocardial perfusion grade determined after percutaneous coronary intervention. Group I consisted of 223 patients with good myocardial perfusion (TMPG 2-3) after successful percutaneous coronary intervention and group II of 60 patients with poor myocardial perfusion (TMPG 0-1). RESULTS: The prevalence of metabolic syndrome was found to be significantly higher in patients with poor myocardial perfusion than in those with good myocardial perfusion (40 vs. 20%, respectively, P=0.002). Moreover, we detected an independent association between metabolic syndrome and the occurrence of poor myocardial perfusion grade (adjusted OR=2.54, 95% CI=1.35-4.75, P=0.003). CONCLUSIONS: We have shown, for the first time, a significant association between metabolic syndrome and impaired myocardial perfusion after percutaneous coronary intervention in patients with acute myocardial infarction. This data may partially explain the poor short and long-term outcomes of acute myocardial infarction in patients with metabolic syndrome. PMID- 16707958 TI - Spatial and phasic oscillation of non-Newtonian wall shear stress in human left coronary artery bifurcation: an insight to atherogenesis. AB - OBJECTIVE: To investigate the wall shear stress oscillation in a normal human left coronary artery bifurcation computational model by applying non-Newtonian blood properties and phasic flow. METHODS: The three-dimensional geometry of the investigated model included the left main coronary artery along with its two main branches, namely the left anterior descending and the left circumflex artery. For the computational analyses a pulsatile non-Newtonian flow was applied. To evaluate the cyclic variations in wall shear stress, six characteristic time points of the cardiac cycle were selected. The non-Newtonian wall shear stress variation was compared with the Newtonian one. RESULTS: The wall shear stress varied remarkably in time and space. The flow divider region encountered higher wall shear stress values than the lateral walls throughout the entire cardiac cycle. The wall shear stress exhibited remarkably lower and oscillatory values in systole as compared with that in diastole in the entire bifurcation region, especially in the lateral walls. Although the Newtonian wall shear stress experienced consistently lower values throughout the entire cardiac cycle than the non-Newtonian wall shear stress, the general pattern of lower wall shear stress values at the lateral walls, particularly during systole, was evident regardless of the blood properties. CONCLUSIONS: The lateral walls of the bifurcation, where low and oscillating wall shear stress is observed, are more susceptible to atherosclerosis. The systolic period, rather than the diastolic one, favors the development and progression of atherosclerosis. The blood viscosity properties do not seem to qualitatively affect the spatial and temporal distribution of the wall shear stress. PMID- 16707957 TI - Increase in circulating bone marrow progenitor cells after myocardial infarction. AB - BACKGROUND: Most circulating blood cells expressing the marker CD34 are bone marrow progenitor cells. These cells differentiate into cardiomyocytes, endothelial and smooth muscle cells after myocardial infarction in vivo. Mobilization of bone marrow progenitor cells into the peripheral blood after myocardial infarction may supply these cells to the heart. Rise in CD34+ cell concentrations following myocardial infarction would support the existence of myocardial-initiated mobilization. METHODS: Serial measurements of circulating CD34+ cells were made in 42 consecutive patients presenting with first ST elevation myocardial infarction. Measurement of serum concentrations of monocyte chemoattractant protein-1, stromal derived factor-1, hepatocyte growth factor, interleukin-17 and thrombopoietin was also performed. Samples were drawn on day 1 after myocardial infarction, and on days 4, 8 and 12. Levels of CD34+ cells and cytokines were also measured in 15 controls. RESULTS: By day 8, the mean concentration of CD34+ cells rose by 74% above mean control level of 2527 cells/ml, and 41% above day 1 mean (P=0.02). This rise was sustained on day 12 (P=0.05). On day 1, there was a 9.3-fold rise in hepatocyte growth factor above the control level of 589 pg/ml (P=0.002). Hepatocyte growth factor levels declined from the day 1 mean of 6061 to 1485 pg/ml on day 12 (P=0.002). No significant change in stromal derived factor-1, interleukin-17, monocyte chemoattractant protein-1 and thrombopoietin was observed. Elevations in CD34+ cells and hepatocyte growth factor were not related to infarction size as estimated on echocardiography. CONCLUSIONS: Elevation in the concentration of circulating CD34+ cells after myocardial infarction suggests that myocardial initiated bone marrow progenitor cell mobilization exists in humans. The cytokines studied in our protocol are not likely to play a direct role in bone marrow progenitor cell mobilization. PMID- 16707959 TI - Assessment of coronary plaque morphology by contrast-enhanced computed tomographic angiography: comparison with intravascular ultrasound. AB - BACKGROUND: Computed tomographic (CT) angiography provides accurate noninvasive assessment for coronary artery stenosis. The ability of CT angiography to determine plaque morphology remains unclear. METHODS: Twelve patients undergoing intravascular ultrasound for clinical indications underwent CT angiography for the evaluation of plaque morphology. Plaque morphology was classified as (1) soft, (2) fibrous, (3) fibrocalcific or (4) calcific. CT angiography data sets were evaluated for the presence and morphology of plaque in the coronary artery segments. The results were blindly compared with intravascular ultrasound results on a segment by segment basis using angiographic landmarks. RESULTS: Fifty-nine coronary segments were analyzed; 10 segments were normal and 49 contained plaque. Plaque morphology by intravascular ultrasound was soft in 12 segments, fibrous in four, fibrocalcific in 29 and calcific in four. To determine coronary segments with any plaque, CT angiography had a sensitivity of 100% (49 of 49) and specificity of 90% (9 of 10). To determine plaque morphology as calcified (fibrocalcific and calcific) versus noncalcified (soft and fibrous), CT angiography had a sensitivity of 100% (33 of 33) and a specificity of 94% (15 of 16). Overall accuracy for CT angiography to determine plaque morphology was 92% (54 of 59). CT angiography density values (Hounsfield units, mean+/-standard deviation) were significantly different for each plaque morphology: soft 23+/-71, fibrous 108+/-79, fibrocalcific 299+/-112 and calcific 404+/-264 (P<0.0001). CONCLUSIONS: CT angiography accurately characterized plaque morphology and may be a useful tool in noninvasive evaluation of plaque morphology during drug therapy trials. PMID- 16707960 TI - Decreased nitrate-mediated dilatation in patients with coronary artery ectasia: an ultrasonographic evaluation of brachial artery. AB - BACKGROUND: Coronary artery ectasia has been defined as localized or diffuse nonobstructive lesions of the epicardial coronary arteries with a luminal dilation exceeding the 1.5-fold of normal adjacent segment or vessel diameter. Although coronary artery disease is supposed to be responsible for more than 50% of coronary ectasia, the precise pathology of coronary artery ectasia is not clearly understood. The brachial artery ultrasound test for flow-mediated endothelial-dependent vasodilatory function includes administration of sublingual nitrates to examine the vasodilating effect of an exogenous source of nitric oxide. In the present study, we aimed to compare flow-mediated and nitrate mediated responses of brachial artery in patients with coronary artery ectasia and patients with coronary artery disease. MATERIALS AND METHODS: Thirty-six consecutive patients with coronary artery ectasia in combination with coronary artery disease and 42 age-matched and sex-matched patients with coronary artery disease alone were included in the study. Flow-mediated and nitrate-mediated dilatations were measured in all patients using a high-resolution B-mode ultrasonographic system. RESULTS: Baseline brachial artery diameters in patients with coronary artery ectasia were not statistically different from those in patients with coronary artery disease (4.2+/-0.6 vs. 4.0+/-0.6 mm, respectively, P=0.16). Although the forearm flow-mediated dilatation of the patients with coronary artery ectasia did not differ from that of patients with coronary artery disease alone (5.5+/-3.8 vs. 4.8+/-3.6%, respectively, P=0.41), nitrate-mediated dilatation was significantly lower than that of patients with coronary artery disease alone (7.9+/-5.2 vs. 10.9+/-5.4%, respectively, P=0.02). CONCLUSION: We have shown that patients with coronary artery ectasia have decreased nitrate mediated response of brachial artery compared with patients with coronary artery disease alone, suggesting more severe dysfunction or, possibly, destruction of the media layer in coronary artery ectasia than in coronary artery disease. PMID- 16707961 TI - Intracoronary radiotherapy with a 188Rhenium liquid-filled angioplasty balloon system in in-stent restenosis: a single-center, prospective, randomized, placebo controlled, double-blind evaluation. AB - BACKGROUND: In cases of in-stent restenosis, intracoronary radiotherapy with beta emitters and gamma-emitters has been shown to reduce the risk of repeat restenosis. The present randomised, placebo-controlled study addresses the question of whether intracoronary radiotherapy applied by the easy-to-handle Rhenium liquid-filled angioplasty balloon system is also able to reduce the angiographic re-restenosis rate in stents. METHODS AND RESULTS: At our center, from May 2000 to December 2003, 165 patients (mean age 64+/-10, median 65 years; 127 men, 38 women) with symptomatic in-stent restenosis underwent either intracoronary brachytherapy or sham procedure. Index clinical and angiographic parameters were largely comparable in both groups. Radiation therapy was performed with a standard percutaneous transluminal coronary angioplasty (PTCA) balloon catheter inflated with liquid Rhenium in the redilated in-stent restenosis for 240-890, mean 384+/-125 s with low pressure (3 atm) in order to reach 30 Gy at 0.5 mm depth of the vessel wall. In 82 patients, intracoronary radiotherapy was carried out without complications, but one of the 83 patients who underwent sham procedure suffered small myocardial infarction. During follow up, stent thrombosis with subsequent non-Q-wave myocardial infarction occurred in one patient in each group (6 days and 8 months after the procedure, respectively). At 6 months after the index procedure, repeat angiography was performed in 156 of the 164 patients with successful procedure (rate 95%): restenosis (stenosis >50% in diameter) or reocclusion was observed in only 19 of 78 (=24%) patients of the radiation but in 31 of 78 (=40%) patients of the sham procedure group (P=0.04). Event-free survival (free of death, myocardial infarction, target vessel revascularization) at 1 year was 87% for patients being radiated and 74% for patients having undergone sham procedure (P=0.05). CONCLUSIONS: Intracoronary radiation therapy with the liquid-filled beta-emitting Rhenium balloon is not only easy to perform, safe, and comparably inexpensive but also an effective option to prevent repeat restenosis and the need for target vessel revascularization in cases of in-stent restenosis. PMID- 16707962 TI - Initial impact of drug-eluting stent use on the spectrum of patients undergoing percutaneous coronary intervention. AB - BACKGROUND: The availability of drug-eluting stents was expected to expand the application of percutaneous coronary intervention to a wider group of patients and lesion types. We sought to determine whether drug-eluting stents' availability has changed the practices of operators performing percutaneous coronary intervention with regard to patient selection and procedural factors. METHODS: We compared the clinical and angiographic characteristics of patients who underwent percutaneous coronary intervention at the Methodist Hospital, Houston, Texas, during three periods: June 2002-March 2003, June 2003-March 2004 and June 2004-March 2005. Sirolimus-eluting stents were available during the latter two periods. Paclitaxel eluting stents became available during the third period. RESULTS: A greater proportion of patients undergoing diagnostic catheterization were referred for percutaneous coronary intervention during the latter two periods (26.8%, 30.4%, 30.4%, respectively, P<0.0001). Patients undergoing stent placement during the latter two periods were more likely to have hypertension, hyperlipidemia or to have undergone previous percutaneous coronary intervention. The average implanted stent length was greater during the latter periods (14.5+/-4.6, 16.4+/-5.2, 16.8+/-5.5 mm, respectively, P<0.0001) and the average stent diameter was smaller (3.23+/-1.22, 3.13+/-1.16, 3.02+/-0.6 mm, respectively, P<0.0001). The frequency of percutaneous coronary intervention involving long lesions (>20 mm) and Left Anterior Descending (LAD) lesions was higher in the latter two periods and the frequency of multivessel stenting was higher in the last period. CONCLUSIONS: The spectrum of patients and coronary lesions that have undergone stenting has changed, particularly in the third period when both drug-eluting stent types were available. We observed a gradual shift toward higher-risk clinical and lesion characteristics following the introduction of drug-eluting stents. PMID- 16707964 TI - Bibliography. Current world literature. PMID- 16707963 TI - Long-term results following percutaneous myocardial laser therapy. AB - OBJECTIVES: The usefulness and safety of percutaneous myocardial laser therapy in selected patients have been identified in previous 1-year randomized trial reports, including that from a double-blind, sham-controlled trial we independently conducted. We aimed to determine whether the 1-year effects are maintained through a long-term, longitudinal follow-up. METHODS: Patients (n=77) with chronic, stable, medically refractory angina (class III or IV) not amenable to conventional revascularization and with evidence of reversible ischemia, ejection fraction > or =25%, and myocardial wall thickness > or =8 mm were treated with percutaneous myocardial laser. After the 1-year follow-up and disclosure of all randomized assignments as prespecified in the respective study protocol, patients were followed up longitudinally for a mean of 3 years for angina class, left ventricular ejection fraction, medication usage, and adverse events. RESULTS: No procedural mortality, myocardial infarction, or cerebral embolism occurred. Pericardiocentesis was required in two patients (2.6%). Cardiac event-free survival was 88% at 1 year and 66% at late follow-up. Mean Canadian Cardiovascular Society angina class was significantly improved from baseline (3.2+/-0.4) at 1 year (2.2+/-1.1, P<0.001) and at a mean of 3 years (1.9+/-1.2, P<0.001). Nitrate usage was significantly reduced at late follow-up; however, ejection fraction did not change over time. In a multivariate analysis, angina improvement at 1 year was found to be a significant independent predictor of both survival and angina improvement at late follow-up. CONCLUSION: We conclude that percutaneous myocardial laser therapy in selected patients with severe, medically refractory angina not treatable with conventional revascularization induces significant and sustained symptomatic benefit. PMID- 16707965 TI - Endoscopic appearance of annular pancreas in a patient with Schinzel-Giedon syndrome. PMID- 16707966 TI - Probiotics in gastrointestinal diseases in children: hard and not-so-hard evidence of efficacy. AB - The use of probiotics, once discussed primarily in the context of alternative medicine, is now entering mainstream medicine. However, only a few of the potential health benefits attributed to probiotics have been confirmed in well designed, well-conducted, randomized, controlled trials. This is especially true in the pediatric population. We review here the available evidence on efficacy of probiotics in children in the prevention and treatment of gastrointestinal diseases. Although we restrict our analysis to the pediatric age, whenever potentially relevant information is available only from adult studies, they are examined as well. Probiotics have been most extensively studied in the treatment of diarrheal diseases, where their efficacy can be considered well established. Studies documenting effects in other childhood gastrointestinal illnesses are few, although some preliminary results are promising. Furthermore, only a limited number of probiotic strains have been tested, and, as the effects of different probiotic microorganisms are not equivalent, results cannot be generalized. Thus, at present, we have some positive certainties, lots of exciting promises and many unanswered questions. PMID- 16707967 TI - The exposure of infants to Lactobacillus rhamnosus GG in Finland. PMID- 16707968 TI - Cytokine tumor necrosis factor-alpha A promoter gene polymorphism at position 308 G-->A and pediatric inflammatory bowel disease: implications in ulcerative colitis and Crohn's disease. AB - OBJECTIVES: Our pilot study aimed to determine the effect of tumor necrosis factor-alpha (TNF-alpha) 308 G-->A promoter single-nucleotide polymorphism in pediatric inflammatory bowel disease (IBD), its influence on inflammatory activity and the clinical manifestations. METHODS: We obtained genomic DNA from 164 subjects, 82 with long-standing IBD aged 8 to 18 years: 46 with Crohn disease (CD) and 36 with ulcerative colitis (UC). Eighty-two healthy children served as the control population. Genotyping was determined by using a restriction enzyme based assay. TNF-alpha 308 G-->A polymorphism was assessed in terms of inflammatory (C-reactive protein [CRP]) and disease activity. The latter was assessed by the Pediatric Crohn's Disease Activity Index (PCDAI) and the Truelove index for CD and UC, respectively. RESULTS: Significant differences in TNF-alpha 308 A polymorphism were found between the IBD group and controls (P < 0.05) and the UC group and controls (P < 0.001). No differences were noted between TNF alpha 308 A polymorphism and clinical characteristics in UC. The frequency of the -308 A allele of TNF was not different in CD compared with that in the control group. The frequency of TNF-alpha 308 A genotype was significantly higher in CD patients with predominantly stenosing/penetrating disease compared with patients without complications (P < 0.001) and healthy controls (P < 0.01). In CD patients, those carrying TNF -308 A had a significant increase in CRP (P < 0.05) and the PCDAI (P < 0.05). In CD, CRP levels strongly correlated with the PCDAI (r = 0.6150, P < 0.001). In UC, significant differences among the mean levels of CRP (P < 0.05) and disease activity (P < 0.001) related to TNF-alpha 308 A polymorphism were found. Allele distribution (odds ratio, 12.9; CI, 1.18-140.81, P < 0.001) and CRP serum levels (odds ratio, 1.020; CI, 1.00-1.04, P < 0.001) were independently associated with CD complications. CONCLUSIONS: Although not necessarily dictating IBD initiation, the TNF-alpha 308 A polymorphism may play a role in modifying the CD phenotype. The polymorphism may influence disease activity as well as more intense inflammatory activity in both forms of IBD and may modify the progression of chronic digestive tract inflammation. PMID- 16707969 TI - Changes of gut microbiota and immune markers during the complementary feeding period in healthy breast-fed infants. AB - INTRODUCTION: Little is known about changes in intestinal microbiota during the important period of complementary feeding (weaning). This descriptive study investigated changes of selected gut microbiota and markers of gut permeability and the immune system in breast fed infants during the complementary feeding period. METHODS: 22 healthy, exclusively breast fed infants (from birth to 4 months) with no antibiotic intake during the month prior to the study, were followed from 4 to 9 months of age. Faecal and saliva samples were collected at the start of the study (V0) and at monthly intervals (V1-V5) for measurement of selective gut microbiota (bifidobacteria, lactobacilli, vancomycin-insensitive lactobacilli, enterobacteria, enterococci, Clostridium perfringens) using semi selective media. Immune markers (alpha-1-antitrypsin, eosinophil cationic protein (ECP), secretory IgA and TNF-alpha were measured in saliva and secretory IgA and TNF-alpha in faecal samples. RESULTS: High stool bifidobacteria counts at the start of the study (7.99 1 1.95 log10 CFU/g faeces) remained stable throughout the 5 months of complementary feeding while counts of enterobacteria and enterococci increased with age (P < 0.05 and P = 0.02 respectively). Vancomycin insensitive lactobacilli increased significantly during weaning for V0 to V3 (P < 0.01), and then decreased slightly (V4). Faecal Clostridium perfringens remained below the detection limit during the study and parameters measured in saliva did not change. Faecal ECP decreased significantly from 1.011.4 (V0) to 0.510.9 mg/mg protein (V5) P = 0.03. CONCLUSION: Age and/or diet modifications during complementary feeding had no impact on faecal bifidobacteria counts but increased those of enterobacteria and enterococci. Transient increases in faecal lactobacilli and vancomycin-insensitive lactobacilli counts were observed. The reduction in faecal ECP may indicate a decrease in gut permeability (reinforcement of gut mucosa integrity) during the weaning period with age [corrected] PMID- 16707970 TI - Diagnostic tests in Hirschsprung disease: a systematic review. AB - OBJECTIVE: We conducted a systematic review to determine and compare the diagnostic accuracy of contrast enema (CE), anorectal manometry (ARM) and rectal suction biopsy (RSB) in infants suspected of Hirschsprung disease. DESIGN: This is a systematic review. DATA SOURCES: Articles were identified through electronic searches in Medline, EMBASE.com and Cochrane Controlled Trials Register. Searches were limited to articles published after 1966 in PubMed and after 1980 in EMBASE.com. STUDY SELECTION: Studies were included if infants underwent at least one of the following tests: CE, ARM or RSB, followed by full-thickness biopsy and/or clinical follow-up as the reference standard. DATA EXTRACTION: Two reviewers independently assessed the methods of data collection, patient selection, blinding and prevention of verification bias and description of the test protocol and reference standard. Data to construct 2 x 2 tables were abstracted for each test. RESULTS: Twenty-four studies met our inclusion criteria, but 2 studies were subsequently excluded for statistical analysis because data was missing to construct the 2 x 2 table. RSB (14 studies for a total of 993 patients) was the most accurate test, having both the highest mean sensitivity (93%; 95% confidence interval [CI], 88%-95%) and mean specificity (98%; 95% CI, 95%-99%). Sensitivity and specificity of ARM (9 studies for a total of 400 patients) were similar to those of RSB (91% vs 93%, P = 0.73 and 94% vs 98%, P = 0.08, respectively). Sensitivity and specificity of CE (12 studies for a total of 425 patients) were significantly lower than those of RSB and ARM, with mean sensitivity and mean specificity of 70% and 83%, respectively. CONCLUSIONS: RSB and ARM are the most accurate tests in the diagnostic workup of Hirschsprung disease. PMID- 16707971 TI - Severity of basal cell hyperplasia differs in reflux versus eosinophilic esophagitis. AB - OBJECTIVES: Basal cell hyperplasia of the esophageal epithelium is a frequent finding in children with histological evidence of esophagitis. The aim of this study was to compare the severity of basal cell hyperplasia in gastroesophageal reflux vs eosinophilic esophagitis. METHODS: A cohort of pediatric patients who underwent same-day endoscopy with esophageal biopsy and 24-hour esophageal pH monitoring was divided into groups based on endoscopic and pH monitoring findings. Basal cell hyperplasia was defined as normal (< or = 25% of esophageal epithelial height), mild (26%-50%), moderate (51%-75%) or severe (> 75%). The severity of basal cell hyperplasia in patients with abnormal pH monitoring studies, both with and without endoscopic abnormalities of the esophagus, was compared with the severity in patients with eosinophilic esophagitis. RESULTS: Twenty-seven children with abnormal pH monitoring were identified. Of these 27 children, 11 had endoscopic findings consistent with reflux esophagitis. Thirty patients with eosinophilic esophagitis were identified. Patients with eosinophilic esophagitis had significantly increased severity (P < 0.001) of basal cell hyperplasia (87% severe, 3% moderate, 3% mild, 7%, normal) than patients with abnormal esophageal pH monitoring alone (11% severe, 4% moderate, 15% mild, 70% normal) or in combination with endoscopic abnormalities (18% severe, 9% moderate, 18% mild, 55% normal). CONCLUSIONS: Basal cell hyperplasia is more severe in children with eosinophilic esophagitis than in those with reflux esophagitis. The finding of basal cell hyperplasia is a powerful clue into the underlying etiology of pediatric esophagitis and, along with epithelial eosinophil count, can be used as information to guide therapy. PMID- 16707972 TI - Dilated intercellular spaces: a major morphological feature of esophagitis. AB - OBJECTIVES: Dilated intercellular spaces (DIS) in the esophageal epithelium have been identified by electron microscopy as marker of acid reflux damage in experimental animals and adults with gastroesophageal reflux disease (GERD). We aimed to identify and quantify DIS by light microscopy in pediatric GERD and esophagitis. METHODS: We prospectively took esophageal biopsies in 70 consecutive pediatric patients, 48 of whom had GERD symptoms. On hematoxylin and eosin stained sections esophagitis was scored histologically, and DIS were graded as 0 (absent), + (small and focal), ++ (moderate) or +++ (large and diffuse). A computerized image analysis identified total, cellular and nuclear areas and DIS were quantified as percentage of total minus cellular area. RESULTS: Forty of 48 GERD patients had histological esophagitis (33 G1, 4 G2, 3 G3, 1 of which with Barrett esophagus), and all 40 had DIS (33 +, 4 ++, 3 +++) with 100% interobserver agreement; 15 of 29 (55%) had abnormal pH study (reflux index, 5.7% 36%). In 30 patients the esophagus was histologically normal. DIS values were 2.21% +/- 2.60% (range, 0.11%-12%) in patients with esophagitis and 0.44% +/- 0.13% (0.2%-0.7%) in patients with normal histology (P < 0.00001), with 0.71% bearing 70% sensitivity and 100% specificity for GERD versus controls. Five other children with esophagitis unrelated to GERD (eosinophilic, Candida, food allergy) also had DIS + to +++, and median DIS area was 5% (1.3%-12%). CONCLUSIONS: DIS can be detected and evaluated by light microscopy, and the image analysis used provides an objective quantification of DIS and supports the light microscopy evaluation. DIS are a morphological feature of GERD and esophagitis in infancy and childhood. PMID- 16707974 TI - Superior mesenteric artery syndrome in children: a 20-year experience. AB - OBJECTIVES: Superior mesenteric artery syndrome (SMAS), Wilkie syndrome or cast syndrome is a rare condition that usually presents with symptoms of mid to upper gastrointestinal obstruction due to the compression of the duodenum between the abdominal aorta, posteriorly, and the superior mesenteric artery, anteriorly. The aim of this study was to analyze the clinical characteristics, means of diagnosis and management of SMAS in a pediatric population. METHODS: Retrospective chart review of all patients at the Children's Hospital of Wisconsin with SMAS from 1985 to 2005. RESULTS: Twenty-two cases of SMAS where diagnosed at Children's Hospital of Wisconsin between 1985 and 2005 [14, (64%) female]. Symptoms developed 1 to 393 days (median 5 days) before diagnosis. Presenting symptoms included abdominal pain (59%), vomiting (50%), nausea (40%), early satiety (32%) and anorexia (18%). Diagnosis was made by upper-gastrointestinal radiography in 18 (82%), by computed tomography in 2 (9%) and at laparotomy in 2 (9%). One patient was treated surgically after medical management failed. Mean length of treatment was 65 days (range 13-169), with a mean length of hospitalization of 21 days (range 0-68 days). CONCLUSION: SMAS usually presents more acutely than chronically with symptoms of small bowel obstruction. Weight loss is not necessary for SMAS development. Prior neurological injury may be a risk factor for development of SMAS. Upper gastrointestinal radiography remains the primary means of diagnosis. SMAS is typically successfully managed medically. Surgical intervention should be reserved for patients' refractory to medical therapy. The expected outcome of SMAS is excellent. PMID- 16707973 TI - Allergic eosinophilic gastroenteritis with protein-losing enteropathy: intestinal pathology, clinical course, and long-term follow-up. AB - OBJECTIVES: A subset of patients with allergic eosinophilic gastroenteritis (AEG) has anemia and hypoalbuminemia caused by protein-losing enteropathy (PLE). Our goals were to describe the response to therapy and the long-term outcome of patients in this subgroup and to evaluate their gastric and intestinal biopsies for distinguishing features that might explain their protein and blood loss. METHODS: Patients with AEG + PLE were identified retrospectively and compared with controls and with patients with AEG only. Immunohistochemical staining for tryptase, a mast cell mediator, was performed on gastric and duodenal tissues. Eosinophils identified by hematoxylin/eosin stain and mast cells identified as tryptase-positive cells were counted in one high-power field area with maximal cell infiltration. RESULTS: Although all patients had excellent response to therapy with amino acid-based formula and tolerated gradual introduction of some foods with time, food-responsive disease persisted in all patients over 2.5 to 5.5 years of follow-up. Routine histological evaluation did not show any features differentiating AEG + PLE from AEG. When eosinophils and mast cells were counted in intestinal biopsies, however, significantly more mast cells were found in biopsies of the AEG + PLE group despite comparable numbers of eosinophils. In contrast, in gastric biopsies, eosinophils were more prominent in AEG + PLE, but mast cell numbers were similar in all groups. CONCLUSIONS: Patients with AEG + PLE responded well to therapy with amino acid-based formula. Food hypersensitivities did not completely resolve over up to 5.5 years. Intestinal mast cells were significantly increased in maximally infiltrated areas of the intestine, possibly causing increased intestinal permeability and protein loss. PMID- 16707975 TI - Endoscopic management of sphincter of Oddi dysfunction in children. AB - BACKGROUND: Data on sphincter of Oddi dysfunction (SOD) in children are scant. Most children diagnosed with SOD are treated by biliary sphincterotomy with suboptimal results. The efficacy and safety of pancreatic and dual sphincterotomy in children with SOD has not been previously reported. OBJECTIVE: To evaluate the efficacy and safety of pancreatic and dual sphincterotomy in children with SOD. MATERIALS AND METHODS: Prospective evaluation of all children who underwent endoscopic retrograde cholangiopancreatogram (ERCP) with sphincter of Oddi manometry for evaluation of suspected SOD over a 3-year period. Children diagnosed with SOD underwent pancreatic or dual sphincterotomy with prophylactic pancreatic stenting. RESULTS: SOD was diagnosed by sphincter of Oddi manometry in 6 of 11 children who underwent ERCP for suspected SOD. Of the 6 children (mean age, 11 years; range, 5-16; 4 girls) with SOD, 3 presented with recurrent pancreatitis and 3 with postcholecystectomy pain. Pancreatic sphincter hypertension was noted in all 6 patients; concomitant biliary sphincter hypertension was noted in 3 patients with postcholecystectomy pain. Patients with recurrent pancreatitis underwent pancreatic sphincterotomy and those with postcholecystectomy pain underwent dual sphincterotomy. Prophylactic pancreatic stents were placed in all patients. One girl experienced mild post-ERCP pancreatitis. At a mean follow-up of 583 days (range, 325-1445), 4 patients were asymptomatic, 1 experienced partial symptom relief and 1 had recurrent symptoms. CONCLUSIONS: As in adults, pancreatic and dual sphincterotomy, in expert hands, is effective and safe in a subgroup of children with SOD. Prospective, randomized trials with larger number of patients are required to validate the efficacy of endotherapy in children with SOD. PMID- 16707976 TI - The "red umbilicus": a diagnostic sign of cow's milk protein intolerance. AB - INTRODUCTION: Red umbilicus is considered to be an infectious disease typical of neonates. In our experience, umbilical erythema could be due to cow's milk protein intolerance (CMPI). AIMS: To evaluate the frequency and clinical significance of umbilical erythema in a series of consecutive children referred for suspected CMPI. PATIENTS AND METHODS: Seven hundred ninety-six consecutive patients (median age, 18 months) referred for suspected CMPI diagnosis were studied. CMPI diagnosis was based on the disappearance of symptoms on elimination diet and their subsequent reappearance on double-blind placebo-controlled cow's milk challenge. RESULTS: CMPI was diagnosed in 384 patients: 120 with respiratory, 75 dermatologic and 198 gastroenterological symptoms. Although some patients showed more than 1 type of symptom, whether gastroenterological, dermatologic or respiratory, they were classified in 1 category only according to the main reason for referral to the outpatients clinic. Umbilical erythema was observed in 36 patients (median age, 10 months): 16 (8%) with gastroenterological symptoms, 9 (7.5%) with recurrent asthma and 11 (15%) with atopic dermatitis. None of the symptomatic controls without CMPI had umbilical erythema. On elimination diet, the erythema disappeared within the second week. On CMPI challenge, it reappeared within 24 hours. CONCLUSIONS: Umbilical erythema can be a sign of food intolerance and can be a useful diagnostic tool for CMPI. PMID- 16707977 TI - 6-Thioguanine-related chronic hepatotoxicity and variceal haemorrhage in children treated for acute lymphoblastic leukaemia--a dual-centre experience. AB - BACKGROUND: 6-Thioguanine treatment in childhood acute lymphoblastic leukaemia (ALL) has been shown to cause hepatic veno-occlusive disease, but this usually resolved with drug withdrawal. Recent reports suggested that treatment of ALL with 6-thioguanine can lead to chronic hepatotoxicity and portal hypertension. We describe our experience from 2 UK centres of chronic hepatotoxicity in children receiving maintenance 6-thioguanine for ALL in the national leukaemia protocol ALL 97/99. METHODS: Retrospective review of children who were referred with liver disease secondary to 6-thioguanine treatment of ALL was performed. A paediatric pathologist blinded to the clinical features reviewed liver histology slides. RESULTS: Ten of 75 children (13%) treated with 6-thioguanine in both centres were referred at a median of 6 months (range, 2-29) after discontinuation of chemotherapy. In 8 cases, referral was due to persistent thrombocytopenia and splenomegaly. Two children presented with acute variceal bleeding. All had thrombocytopenia at referral, and ultrasonography showed coarse hepatic echo texture and splenomegaly in all. Endoscopy showed oesophageal varices in 7 and gastric varices in 1. Nine underwent liver biopsy that showed features compatible with nodular regenerative hyperplasia in 5 cases. After a median follow-up of 36 months, a further child has had a variceal haemorrhage and all but 2 children remain thrombocytopenic. CONCLUSIONS: 6-Thioguanine-induced chronic hepatotoxicity is a significant complication in children treated with this agent for ALL. Children may present several months to years after discontinuation of 6 thioguanine. All children given maintenance treatment of ALL with this agent should be screened, and affected children require long-term surveillance. PMID- 16707978 TI - Usefulness of nonbreath-hold 1-shot magnetic resonance cholangiopancreatography for the evaluation of choledochal cyst in children. AB - OBJECTIVE: The aim of this study was to clarify the usefulness of magnetic resonance cholangiopancreatography (MRCP) for the evaluation of choledochal cyst in children. SUBJECTS AND METHODS: MRCP was performed preoperatively in 33 patients. The MRCP findings were compared with those of endoscopic retrograde cholangiopancreatography or intraoperative cholangiopancreatography. RESULTS: In all 33 patients, MRCP could detect choledochal cyst. The detection rate of a cyst in the main pancreatic duct was 62.2%, of abnormal union of the pancreaticobiliary junction (AUPBJ) was 53.3%, of dilatation or abnormalities of the main pancreatic duct was 75.0% and of a protein plug or stone was 76.9%. In patients under 2 years of age (group A), these findings were significantly lower than those of patients above 2 years of age (group B) [main pancreatic duct: 16.6% (1/6) vs 73.1% (19/26), P < 0.01; AUPBJ: 0.0% (0/6) vs 66.7% (16/24), P < 0.05; and protein plug or stone: 0.0% (0/2) vs 90.9% (10/11), P < 0.05]. The detection rate of AUPBJ in the patients with fusiform dilatation was superior to that of those with cystic dilatation [70% (14/20) vs 20% (2/10), P < 0.05]. In the patients with fusiform dilatation, the detection rate in group A was significantly lower than that in group B [0.0% (0/3) vs 82.4% (14/17), P < 0.01]; however, there was no significant difference between the 2 groups in the detection of cystic dilatation. CONCLUSION: In patients older than 2 years, MRCP should be the first-choice method for confirming the diagnosis and for ensuring accurate visualization of the pancreaticobiliary system. PMID- 16707979 TI - Lactobacillus rhamnosus GG decreases lipopolysaccharide-induced systemic inflammation in a gastrostomy-fed infant rat model. AB - OBJECTIVES: A gastrostomy-fed rat infant "pup-in-a-cup" model was used to test the hypothesis that enterally administered Lactobacillus rhamnosus GG (LGG) decreases the proinflammatory response induced by Escherichia coli lipopolysaccharide (LPS) in the developing infant rat small intestine, plasma, lung and liver. METHODS: Two groups of 6- to 7-day-old pups were fed a rat milk substitute with LPS added via the gastrostomy tube for 6 days. One of the rat milk substitute-fed groups received supplemental LGG; another group received LPS without LGG. Age-matched mother-fed rat pups were used as controls. RESULTS: LPS treatment blunted body growth, but LGG supplementation had no effect on weight increments. LGG decreased LPS-induced inflammation in intestinal tissue; CINC-1 (rodent IL-8 equivalent) production in plasma, liver, lung and distal small intestine; and tumor necrosis factor alpha (TNF-alpha) production in plasma and lung. Cytokine multiplex assay showed lung interleukin (IL)-1beta, IL-6, IL-10, IL-18, growth-related oncogene (GRO)/KC (rat CINC-1) and TNF-alpha were significantly higher in gastrostomy-fed, LPS-treated pups than in mother-reared pups, and LGG significantly blunted the LPS-induced elevation of IL-1beta, IL-10, IL-18, GRO/KC and TNF-alpha; liver GRO/KC was significantly higher in gastrostomy fed, LPS-treated pups than in mother-reared pups, and LGG significantly blunted the LPS-induced elevation of GRO/KC. CONCLUSIONS: LGG provided by the enteral route is able to downregulate LPS-induced proinflammatory mediators. This effect is not only present in the splanchnic organs, that is, the intestine and the liver, but extends to the plasma and a distal organ, the lung. PMID- 16707980 TI - Bifidogenic effects of solid weaning foods with added prebiotic oligosaccharides: a randomised controlled clinical trial. AB - OBJECTIVES: Breast-fed infants have relatively high proportions of faecal bifidobacteria. This bifidogenic microbiota is at least partly explained by the high levels of prebiotic oligosaccharides in human milk. The present study aimed at testing the effect of solid foods with added prebiotic galacto- and fructo oligosaccharides (GOS/FOS) on the composition of the intestinal microbiota of fully formula-fed infants during the weaning period. METHODS: The study was a double-blind, randomised trial with an intervention period of 6 weeks. Infants aged 4 to 6 months who were about to start consuming solid foods were randomised to receive either weaning products with a mixture of GOS/FOS or control weaning products with maltodextrin. After an adjustment period, the presented daily dose of GOS/FOS was 4.5 g. Total numbers of bacteria and bifidobacteria in faecal samples were analysed with fluorescent in situ hybridization. RESULTS: Thirty five infants were included in the study, and 20 infants were included in a per protocol analysis. In the GOS/FOS group, the faecal percentage of bifidobacteria significantly increased from 43% to 57% (P = 0.031) from week 0 to week 6, but did not significantly change in the control group (36% and 32%, respectively; P = 0.387). The change in the percentage of bifidobacteria between week 0 and week 6 in the GOS/FOS was significantly different from this change in the control group (P = 0.026). CONCLUSIONS: We conclude that addition of GOS/FOS to solid foods induces an increase in the faecal proportion of bifidobacteria in the intestinal microbiota of fully formula-fed infants with an established, mixed-type microbiota in the weaning period. PMID- 16707982 TI - Breast-feeding duration and childhood acute leukemia and lymphomas in a sample of Turkish children. AB - OBJECTIVES: Whether breast-feeding is associated with decreased incidence of the lymphoid malignancies in children is uncertain. We evaluated childhood acute leukemia and lymphoma in relation to duration of breast-feeding. METHODS: We investigated this issue in a case-control study comprising 137 patients, aged 1 to 16 years, with acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML), Hodgkin or non-Hodgkin lymphoma, in addition to 146 controls matched for age and sex. RESULTS: The median duration of breast-feeding among patients was shorter than that of controls (10 vs 12 months). Patients with ALL and AML had shorter mean breast-feeding duration compared with healthy children (P = 0.001 and P < 0.001, respectively). The shortest mean breast-feeding duration was noted in the children with AML. Breast-feeding for a duration of 0 to 6 months, when compared with feeding of longer than 6 months, was associated with increased odds ratios (ORs) for ALL [OR = 2.44, 95% confidence interval (CI) = 1.17-5.10], AML (OR = 6.67, 95% CI = 1.32-33.69), Hodgkin lymphoma (OR = 3.33, 95% CI = 0.60-18.54), non-Hodgkin lymphoma (OR = 1.90, 95% CI = 0.68-5.34) and overall (OR = 2.54, 95% CI = 1.51-4.26). CONCLUSIONS: Our findings suggest that breast-feeding of more than 6 months is protective against childhood lymphoid malignancies, especially for AML and ALL. PMID- 16707981 TI - Anti-rotavirus antibodies in human milk: quantification and neutralizing activity. AB - OBJECTIVE: To analyze anti-rotavirus antibodies in human milk in order to determine their isotypes and neutralizing activity on rotavirus strains representing different viral serotypes. METHODS: One hundred seventy-three milk samples (65 colostrum, 55 transitional milk and 53 mature milk) obtained from 65 mothers were analyzed along with 49 serum samples collected just before delivery. Total immunoglobulin A (IgA) and rotavirus-specific IgA and immunoglobulins G (IgG) antibodies were determined in milk and serum by enzyme-linked immunosorbent assay. Neutralizing activity was evaluated by an immunoperoxidase focus reduction assay. Milk IgA was purified by binding to the lectin jacalin, elution and ultrafiltration. RESULTS: Total IgA antibodies were detected in all serum and milk samples analyzed. IgA levels decreased sharply during the replacement of colostrum by transitional milk, and more gradually from transitional to mature milk. These differences in IgA concentration during the 3 periods were statistically significant. Anti-rotavirus antibodies detected in human milk were exclusively of the IgA type, whereas both IgA and IgG anti-rotavirus antibodies were present in serum samples. Both milk and serum samples showed in vitro neutralization of the infectivity of rotavirus strains SA11, Wa and VA70, this activity being stronger toward the human rotavirus strain Wa. No correlation was however found between the inhibitory effect on rotavirus and the concentrations of IgA in human milk and serum samples. CONCLUSION: Anti-rotavirus antibodies are only partly responsible for the neutralizing activity detected in milk and serum. This result suggests that other components possessing suppressive activity against rotavirus must also be present. PMID- 16707983 TI - Dihydroergotamine-associated intestinal ischemia in a child with cyclic vomiting syndrome. PMID- 16707984 TI - Long-term outcome of a patient with congenital short bowel syndrome. PMID- 16707985 TI - Life-threatening rectal bleeding with Klippel-Trenaunay syndrome controlled by angiographic embolization and rectal clips. PMID- 16707986 TI - Treatment of multiple small bowel angiodysplasias causing severe life-threatening bleeding with thalidomide. PMID- 16707987 TI - Comment on gastrointestinal symptoms associated with orthostatic intolerance. PMID- 16707988 TI - Infliximab for induction and maintenance therapy for ulcerative colitis. PMID- 16707989 TI - Cholesterol intake, biosynthesis of cholesterol and plasma lipids in infants. Effects of early cholesterol intake on cholesterol biosynthesis and plasma lipids among infants until 18 months of age. PMID- 16707990 TI - Do parents help their children? Consumerism in health care can be detrimental to child health: lessons from children with functional abdominal pain. PMID- 16707991 TI - A follow-up study of nutrient intake, nutritional status, and growth in infants with cow milk allergy fed either a soy formula or an extensively hydrolysed whey formula. PMID- 16707992 TI - Feeding preterm infants after hospital discharge: a commentary by the ESPGHAN Committee on Nutrition. AB - Survival of small premature infants has markedly improved during the last few decades. These infants are discharged from hospital care with body weight below the usual birth weight of healthy term infants. Early nutrition support of preterm infants influences long-term health outcomes. Therefore, the ESPGHAN Committee on Nutrition has reviewed available evidence on feeding preterm infants after hospital discharge. Close monitoring of growth during hospital stay and after discharge is recommended to enable the provision of adequate nutrition support. Measurements of length and head circumference, in addition to weight, must be used to identify those preterm infants with poor growth that may need additional nutrition support. Infants with an appropriate weight for postconceptional age at discharge should be breast-fed when possible. When formula-fed, such infants should be fed regular infant formula with provision of long-chain polyunsaturated fatty acids. Infants discharged with a subnormal weight for postconceptional age are at increased risk of long-term growth failure, and the human milk they consume should be supplemented, for example, with a human milk fortifier to provide an adequate nutrient supply. If formula fed, such infants should receive special postdischarge formula with high contents of protein, minerals and trace elements as well as an long-chain polyunsaturated fatty acid supply, at least until a postconceptional age of 40 weeks, but possibly until about 52 weeks postconceptional age. Continued growth monitoring is required to adapt feeding choices to the needs of individual infants and to avoid underfeeding or overfeeding. PMID- 16707993 TI - Probiotic intervention in neonates--will permanent colonization ensue? AB - The presence of Lactobacillus rhamnosus GG at 6 and 12 months in the feces of 128 infants receiving the strain or placebo for 6 months after birth was monitored. No permanent establishment of the strain was found, but transient colonization was observed at 6 months of age. The presence of the strain at the end of the administration period was correlated with a reduced prevalence of atopic eczema later in life. PMID- 16707994 TI - Clinical quiz. Cytomegalovirus infection. PMID- 16707996 TI - Mental health parity, access, and quality of care. PMID- 16707997 TI - The effects of state parity laws on the use of mental health care. AB - OBJECTIVE: We used a quasiexperimental research design to measure the effect of state parity laws on the use of mental health care in the past year. METHODS: We pooled cross-sectional data from the 2001, 2002, and 2003 National Surveys on Drug Use and Health. Our sample included 83,531 adults 18 years of age or over with private health insurance stratified by the level of mental and emotional distress experienced in the worst month of the past year. We used a state and year-fixed effects approach to measure the effect of parity. Similar to a difference-in-difference analysis, the effect of parity was measured by comparing pre-/postchanges in mental health service use within states that switched active parity status to changes in service use within states that did not change parity status in the same calendar year. For each subgroup, we report predictions of the percentage point change in any mental health care use, prescription drug use, and outpatient care use resulting from parity laws. RESULTS: Depending on the time window used to define active parity status, we found that parity increased the probability of using any mental health care in the past year by as much as 1.2 percentage points (P<0.01) for the lower distress group and by as much as 1.8 percentage points (P<0.05) in the middle distress group. We found no statistically significant changes in service use for the upper distress group. Whether measured differences were attributable to changes in the use of prescription drug or outpatient care also depended on the definition of active parity status. CONCLUSIONS: Overall, the results of this study suggest that state parity laws succeeded in expanding access to mental health care for those with relatively mild mental health problems. PMID- 16707999 TI - Mammography self-report and mammography claims: racial, ethnic, and socioeconomic discrepancies among elderly women. AB - BACKGROUND: National self-report surveys show minimal racial disparity in mammography, whereas analyses of administrative data show large disparity. METHODS: Using the 1998-2002 Medicare Current Beneficiary Surveys, which contain participants' self-report and claims data, we developed multivariable adjusted models examining factors associated with self-reported mammography and self reported mammography verified by billing records. RESULTS: No racial/ethnic disparities were found in self-reported mammography. Verified mammography, however, revealed significant disparities for race, education, income, insurance, and health status. CONCLUSIONS: Race, education, income, insurance, and health status are associated with a lower likelihood of self-reported mammography verified by the existence of claims data. These data caution against exclusive reliance on self-report survey data to assess disparity in mammography. PMID- 16707998 TI - The impact of parity on major depression treatment quality in the Federal Employees' Health Benefits Program after parity implementation. AB - BACKGROUND: Since the 1990s, parity laws have been implemented to reduce inequities in mental health coverage compared with that for general medical conditions. It is unclear if parity under managed care is associated with improvements in mental health treatment quality. Major depressive disorder (MDD) is a prevalent but often undetected and undertreated and thus could potentially benefit from parity implementation. OBJECTIVE: The objective of this study was to examine the association between parity implementation and changes in MDD treatment quality in the Federal Employees' Health Benefits (FEHB) Program. METHODS: We conducted retrospective analyses of insurance claims data. Logistic regression models estimated quality changes for MDD-diagnosed enrollees from pre- to postparity. SUBJECTS: Subjects included MDD-diagnosed FEHB insured enrollees, aged 18-64, across multiple states and 6 FEHB plans before (1999-2000) and after (2001-2002) parity implementation. MEASURES: Measures included receipt of any antidepressant or psychotherapy within a given calendar year of diagnosis; receipt of appropriate psychotherapy frequency/intensity and duration; and pharmacotherapy duration during acute-phase treatment episodes. RESULTS: Postparity, several plans improved significantly in the likelihood of receiving antidepressant medication. In the acute-phase episodes, the greatest improvement was seen in the likelihood of follow up >or=4 months. Few or no other changes were observed in the acute-phase treatment intensity or duration quality measures. CONCLUSIONS: Parity under managed care was associated with modest improvements. The observed improvements were consistent with secular trends in MDD treatment. Whereas mental health parity is an important policy goal, these results highlight its limitations: improving the financing of care may not be sufficient to improve quality. PMID- 16708000 TI - Veterans Health Administration patients' use of the private sector for coronary revascularization in New York: opportunities to improve outcomes by directing care to high-performance hospitals. AB - OBJECTIVE: We sought to quantify Veterans Health Administration (VA) patients' utilization of coronary revascularization in the private sector and to assess the potential impact of directing this care to high-performance hospitals. METHODS: Using VA and New York State administrative and clinical databases, we conducted a retrospective cohort study examining residents of New York State who were enrolled in the VA and underwent either coronary artery bypass graft (CABG) surgery or percutaneous coronary intervention (PCI) in 1999 or 2000 (n=6562) in either the VA or the private sector. We first calculated the proportion of revascularizations obtained in the VA and the private sector. We then identified the private sector hospitals in which these men obtained revascularizations and determined potential changes in mortality and travel burden associated with directing private sector care to high performance hospitals. RESULTS: VA patients in New York were much more likely to undergo revascularization in the private sector than in VA hospitals: 83% of CABGs (2341/2829) and 87% of PCIs (4054/4665) were obtained in the private sector. Private sector utilization was distributed evenly across high- and low-mortality hospitals. Directing private-sector CABG surgery to high-performance hospitals could have reduced expected mortality by 24% (from 2.3% to 1.7%) and would only increase median travel time from 21 to 30 minutes. The benefit of redirecting PCI care is minimal. CONCLUSIONS: For high mortality procedures that veterans frequently obtain in the private sector, like CABG, directing care to high-performance hospitals may be an effective way to improve outcomes for veterans. PMID- 16708001 TI - Did postoperative mortality increase after the implementation of the Medicare Balanced Budget Act? AB - BACKGROUND: The Balanced Budget Act (BBA) of 1997 was a cost-saving measure designed to reduce Medicare reimbursements by $116.4 billion from 1998 to 2002. Resulting financial strain could adversely affect the quality of patient care in hospitals. OBJECTIVE: We sought to determine whether 30-day mortality rates for surgical patients who developed complications changed at different rates in hospitals under different levels of financial strain from the BBA. METHODS: Pennsylvania hospital discharge data, financial data, and death certificate data from 1997 to 2001 were obtained. A retrospective multivariate analysis examined whether 30-day mortality rates from 8 postoperative complications varied based on degree of hospital financial strain. RESULTS: The average magnitude of Medicare payment reduction on overall hospital net revenues was estimated at 1.8% for hospitals with low BBA impact and 3.5% for hospitals with high impact in 1998, worsening to 2.0% and 4.8%, respectively, by 2001. Mortality rates changed at similar rates for high- and low-impact hospitals from 1997 to 1999, but from 1997 to 2000 mortality rates increased more among patients in high-impact compared with low-impact hospitals (P<0.05). From 2000 to 2001, mortality rates among impact groups converged. There were no statistically significant differences based on BBA impact in changes in nursing staff or length of stay. CONCLUSIONS: The mortality of surgical patients who developed postoperative complications increased to a greater degree in the short term in hospitals affected more by BBA. Measuring the quality impact of reimbursement cuts is necessary to understand cost-quality tradeoffs that may accompany cost-saving reforms. PMID- 16708002 TI - Potential drug-drug interactions in the outpatient setting. AB - BACKGROUND: Although medication safety research has tended to focus on inpatients, the safety of drug use among outpatients is also a concern. OBJECTIVE: We estimate the frequency of potentially interacting concomitant medication dispensing among outpatients. RESEARCH DESIGN: We report the number and percent of patients annually dispensed an object drug of interest (ie, warfarin, digoxin, cyclosporine, or lovastatin/simvastatin) with a potentially interacting drug among a random sample of insured adults receiving care from 10 integrated delivery systems. We use 2 definitions of concomitant dispensing: medications dispensed: 1) during the time period for which the patient had the other medication available ('days supply') and 2) on the same day. We also estimate the number of insured U.S. population codispensed these medication pairs. RESULTS: Among patients dispensed a drug of interest, between 17.8% (95% confidence interval [CI]=17.1-18.6%) and 28.0% (95% CI=24.0-32.1%) were concomitantly dispensed a potentially interacting drug using the "days supply" definition, and between 7.1% (95% CI=6.6-7.7%) and 17.7% (95% CI=14.4-21.1%) using the "same day" definition. Extrapolating to the insured U.S. population, between 1.29 (95% CI=1.25-1.33; same day) and 2.67 (95% CI=2.62-2.72; days supply) million insured adults are dispensed 1 of these 4 potentially interacting pairs. CONCLUSIONS: We found evidence of potentially interacting concomitant medication dispensing among outpatients. An opportunity exists to better understand how such dispensing translates into adverse events and ultimately to improved medication safety. PMID- 16708003 TI - Cluster randomized controlled trial of the effectiveness of audit and feedback and educational outreach on improving nursing practice and patient outcomes. AB - BACKGROUND: Current understanding of implementation methods is limited, and research has focused on changing doctors' behaviors. AIM: Our aim was to evaluate the impact of audit and feedback and educational outreach in improving nursing practice and patient outcomes. METHODS: Using a factorial design, cluster randomized controlled trial, we evaluated 194 community nurses in 157 family practices and 1078 patients with diagnosis of urinary incontinence (UI) for nurses compliance with evidence-linked review criteria for the assessment and management of UI and impact on psychologic and social well-being and symptoms. In the outreach arms, nurses' self-reported barriers informed development of tailored strategies. RESULTS: In comparison with educational materials alone, the implementation methods tested did not improve care at 6 months follow-up. Moderate rates of improvement (10-17% of patients) in performance for the assessment of UI and greater rates of improvement (20-30% of patients) for the management of care were found but effects were similar across arms. Improvement in patient outcomes was more consistently positive for educational outreach than for audit and feedback but differences were not significant. Adjustment for caseload size, severity or duration of UI and patients' age did not alter results. CONCLUSIONS: Printed educational materials alone may be as effective as audit and feedback and educational outreach in improving nurses' performance and outcomes of care for people with UI. Greater understanding of the underlying processes in improving performance within multidisciplinary teams through further, theory-driven studies with "no intervention" control groups and longer follow-up are needed. PMID- 16708004 TI - Lack of association between mortality and timing of surgical fixation in elderly patients with hip fracture: results of a retrospective population-based cohort study. AB - BACKGROUND: Conventional wisdom suggests high-quality care for most patients with hip fractures is surgical fixation within 24 hours to reduce mortality and complications, although there is little evidence to support this standard. OBJECTIVES: We sought to determine the relationship between timing of hip fracture surgery and early mortality. DESIGN AND SUBJECTS: This was a retrospective population-based cohort study of 3981 patients with hip fractures>60 years of age that were admitted to hospitals in one Canadian health region from 1994-2000. METHODS: We collected sociodemographic, prefracture comorbidity, and postoperative complication data. Timing of surgery was classified as within 24 hours ("early surgery," the referent group for all analyses), 24-48 hours, and beyond 48 hours. Main outcome was in-hospital mortality. We used multivariable logistic regression methods, including adjustments with propensity scores and a validated hip fracture-specific mortality index, to determine the independent association between early versus later surgery and mortality. RESULTS: Median age of patients was 82 years, 71% were women, and 26% had >4 prefracture comorbidities. Unadjusted in-hospital mortality was 6%; it was 5% for those who had surgery within 24 hours or from 24 to 48 hours, 10% for surgery beyond 48 hours, and 21% for patients that did not have surgery. Compared with those who had surgery within 24 hours, there was no independent association between timing of surgery and in-hospital mortality (24 48 hours, adjusted odds ratio 0.89, 95% confidence interval 0.62-1.30, P=0.55; beyond 48 hours 1.30, 95% confidence interval 0.86-2.00], P=0.21). CONCLUSIONS: The timing of surgical fixation of hip fracture was not associated with early mortality in carefully adjusted analyses, and the use of "surgery within 24 hours" as a measure of high quality care may be inappropriate. PMID- 16708005 TI - Fragmentation of care for frequently hospitalized urban residents. AB - BACKGROUND: Fragmentation across sites of care may impede efficient healthcare delivery. OBJECTIVES: The objectives of this study were to evaluate fragmentation of hospital care for chronically ill New York City (NYC) residents and its association with enrollment in the New York State (NYS) Medicaid program. RESEARCH DESIGN: We conducted a cross-sectional study using the NYS Department of Health's Statewide Planning and Research Cooperative System discharge database. We identified 53,031 NYC residents admitted 3 or more times to acute care hospitals between 2000 and 2002 with the same principal diagnosis of a specific chronic illness (diabetes, sickle cell anemia, psychosis, substance abuse, cancer, gastrointestinal disease, chronic obstructive pulmonary disease/asthma, coronary artery disease, or congestive heart failure). We also evaluated a larger cohort of 225,421 patients with >or=3 admissions for a specific chronic illness coded as either the principal or a secondary diagnosis. A generalized logit model was used to examine the relationship between fragmentation and each patient's primary insurance adjusted for diagnosis and demographic characteristics. MEASURES: Fragmentation was characterized as high, moderate, or low based on the number of distinct hospitals a patient visited relative to the patient's total number of hospitalizations over the 3-year interval. RESULTS: Among frequently hospitalized NYC residents with select chronic conditions, 17.1% experienced highly fragmented care. This rate was 9.9% for patients with commercial insurance, 24.4% for those with Medicaid, and 9.7% for those with Medicare. The unadjusted odds ratio describing high fragmentation of Medicaid enrollees compared with commercially insured patients was 3.82 (95% confidence interval [CI], 3.50-4.18) and, although attenuated, remained significant after adjustment for demographic characteristics (odds ratio, 1.33; 95% CI, 1.20-1.47). The strongest predictor of fragmentation was a diagnosis of psychosis (OR, 2.81; 95% CI, 2.43-3.25) or substance abuse (OR, 7.58; 95% CI, 6.55-8.77). CONCLUSIONS: In NYC, Medicaid enrollment is associated with greater fragmentation of hospital care, but this is largely attributable to the preponderance of Medicaid enrollees with diagnoses of psychosis and substance abuse. Strategies to improve the efficiency of healthcare delivery should focus on patients with mental illness who are frequently admitted to general hospitals. PMID- 16708006 TI - Development and validation of a psychiatric case-mix system. AB - BACKGROUND: Although difficulties in applying risk-adjustment measures to mental health populations are increasingly evident, a model designed specifically for patients with psychiatric disorders has never been developed. OBJECTIVE: Our objective was to develop and validate a case-mix classification system, the "PsyCMS," for predicting concurrent and future mental health (MH) and substance abuse (SA) healthcare costs and utilization. SUBJECTS: Subjects included 914,225 veterans who used Veterans Administration (VA) healthcare services during fiscal year 1999 (FY99) with any MH/SA diagnosis (International Classification of Diseases, 9th Revision, Clinical Modification [ICD-9-CM] codes 290.00-312.99, 316.00-316.99). METHODS: We derived diagnostic categories from ICD-CM codes using Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition definitions, clinical input, and empiric analyses. Weighted least-squares regression models were developed for concurrent (FY99) and prospective (FY00) MH/SA costs and utilization. We compared the predictive ability of the PsyCMS with several case-mix systems, including adjusted clinical groups, diagnostic cost groups, and the chronic illness and disability payment system. Model performance was evaluated using R-squares and mean absolute prediction errors (MAPEs). RESULTS: Patients with MH/SA diagnoses comprised 29.6% of individuals seen in the VA during FY99. The PsyCMS accounted for a distinct proportion of the variance in concurrent and prospective MH/SA costs (R=0.11 and 0.06, respectively), outpatient MH/SA utilization (R=0.25 and 0.07), and inpatient MH/SA utilization (R=0.13 and 0.05). The PsyCMS performed better than other case mix systems examined with slightly higher R-squares and lower MAPEs. CONCLUSIONS: The PsyCMS has clinically meaningful categories, demonstrates good predictive ability for modeling concurrent and prospective MH/SA costs and utilization, and thus represents a useful method for predicting mental health costs and utilization. PMID- 16708007 TI - Disaster and subsequent healthcare utilization: a longitudinal study among victims, their family members, and control subjects. AB - BACKGROUND: The impact of disasters on primary healthcare utilization is largely unknown. Moreover, it is often overlooked how disaster affects those closest to the primary victims, their family members. OBJECTIVE: The objective of this study was to examine the long-term effects of a catastrophic fire on primary healthcare utilization. RESEARCH DESIGN: We conducted a prospective, population-based cohort study covering 1 year pre- and 3 years postfire. Utilization data were extracted from primary care records. SUBJECTS: Subjects consisted of 286 disaster victims, 802 family members of disaster victims, 3722 community control subjects, and 10,230 patients from a national reference population. MEASURES: As outcome measures, we studied 1) the annual number of contacts in primary care and 2) the annual number of contacts for problems related to mental health. Determinants are injury characteristics of victims and bereavement. All analyses control for age, gender, and insurance status. RESULTS: Being an uninjured victim who witnessed the disaster increases the number of contacts by a factor of 1.55 during the first year postfire (95% confidence interval [CI], 1.35-1.78). Uninjured victims contact the family practitioner more often for mental health-related problems than adolescent community control subjects (incidence rate ratio [IRR], 4.54; 95% CI, 1.69-12.20). In adult family members, the loss of a child predicts overall utilization (IRR, 1.88; 95% CI, 1.35-2.63) and utilization for mental health (IRR, 8.69; 95% CI, 2.10-35.92) during the first year postfire. CONCLUSION: Attention should be paid to the primary care needs of bereaved individuals and those who have witnessed the disaster. PMID- 16708008 TI - Use of guideline recommended follow-up care in cancer survivors: routine or diagnostic indications? AB - BACKGROUND: After potentially curative cancer treatment, patients may receive procedures for routine monitoring for recurrence or for evaluation of symptoms or signs. OBJECTIVE: We sought to characterize surveillance care guideline recommended and other procedures performed in cancer survivors according to routine versus diagnostic indications. METHODS: This was a retrospective cohort study of paper and electronic medical records between 1990 and 2000 from a large midwestern U.S. integrated health care delivery system of 500 patients who received curative treatment of breast, colorectal, endometrial, lung, or prostate cancer. Our measures were the indications for potential surveillance procedures as recommended by clinical practice guidelines or otherwise. RESULTS: Among 14,670 procedures of interest received, 82.0% were performed for routine surveillance, whereas 10.6% were performed for diagnostic indications and 7.3% had indeterminate indications. Office visits most were often delivered for routine indications (91.6%), followed by guideline recommended tests for local recurrence (range 74.1-98.4%, depending on the specific test and cancer). In general, tests that were not recommended in established guidelines were for the purposes of detection of metastatic recurrence and were less often delivered for routine indications (overall frequency 59.2%, P<0.0001 compared with recommended testing). CONCLUSION: Office visits and testing for local recurrence of cancer generally are performed for routine surveillance, regardless of recommendation by practice guidelines. Because procedures not recommended by practice guidelines were more often for diagnostic purposes, classification of patients as undergoing intensive surveillance may be misleading and may require record review to confirm. PMID- 16708009 TI - Barriers to care among American Indians in public health care programs. AB - OBJECTIVE: We sought to examine the extent to which reported barriers to health care services differ between American Indians (AIs) and non-Hispanic Whites (Whites). METHODS: A statewide stratified random sample of Minnesota health care program enrollees was surveyed. Responses from AI and White adult enrollees (n=1281) and parents of child enrollees (n=572) were analyzed using logistic regression models that account for the complex sample design. Barriers examined include: financial, access, and cultural barriers, confidence/trust in providers, and discrimination. RESULTS: Both AIs and Whites report barriers to health care access. However, a greater proportion of AIs report barriers in most categories. Among adults, AIs are more likely to report racial discrimination, cultural misunderstandings, family/work responsibilities, and transportation difficulties, whereas Whites are more likely to report being unable to see their preferred doctor. A higher proportion of adult enrollees compared with parents of child enrollees report barriers in most categories; however, differences between parents of AIs and White children are more substantial. In addition to racial discrimination and cultural misunderstandings, parents of AI children are more likely than parents of White enrollees to report limited clinic hours, lack of respect for religious beliefs, and mistrust of their child's provider as barriers. CONCLUSIONS: Although individuals have enrolled in health care programs and have access to care, barriers to using these services remain. Significant differences between AIs and Whites involve issues of trust, respect, and discrimination. Providers must address barriers experienced by AIs to improve accessibility, acceptability, and quality of care for AI health care consumers. PMID- 16708010 TI - Substance abuse treatment duration for Medicaid versus commercial clients in a health maintenance organization. AB - BACKGROUND: As Medicaid clients have come to be enrolled in managed care, concerns have arisen about the ability of private sector systems to meet the needs of enrollees with substance abuse problems. OBJECTIVES: This project describes treatment initiation and duration for Medicaid and commercial substance abuse treatment clients in a large health maintenance organization (HMO). RESEARCH DESIGN: This study was a prospective secondary analysis of information from HMO databases. Subjects were 641 adult Medicaid clients who contacted the HMO's addiction medicine department in 1996 or 1997 and commercial HMO addiction medicine patients (n=447). First contact with addiction medicine during the study period was the index event. MEASURES: Chief dependent variables were initiation and duration of substance abuse treatment after the index event. RESULTS: Logistic regression showed that longer HMO enrollment predicted treatment initiation after substance abuse assessment, but Medicaid status was not a significant predictor. A competing risks analysis using Cox proportional hazards models indicated that once subjects had initiated, Medicaid was not significantly related to exit from substance abuse treatment. Analysis of health plan disenrollment by Medicaid clients indicated that the most common reason was loss of Medicaid eligibility. CONCLUSIONS: These results raise the possibility that state Medicaid policies may make it difficult for clients to obtain suitable chemical dependency treatment services. PMID- 16708011 TI - Calpain inhibitors delay injury-induced apoptosis in adult mouse spinal cord motor neurons. AB - Here, we investigated the effect of calpain inhibitors on apoptosis in organotypic adult spinal cord slices from mice. An increase in calpain I immunoreactivity was found in the nuclei of motor neurons from slices cultured for 30 min. After 4 h, the immunopositive motor neurons exhibited apoptotic changes including nuclear and chromatin condensation. Eight hours after excision, most motor neurons showed nuclear apoptotic features. Two calpain inhibitors, leupeptin and calpain inhibitor XI, inhibited apoptosis in the motor neurons while the caspase inhibitor Z-VAD.fmk had no effect. Leupeptin, but not calpain inhibitor XI and Z-VAD.fmk, also inhibited nucleosomal DNA fragmentation. These results suggest the involvement of calpain I in the induction of apoptosis in motor neurons of adult spinal cord and that apoptosis can be triggered independent of caspase activation. PMID- 16708012 TI - Characterization of proliferating cells from newborn mouse cochleae. AB - Loss of hair cells in mammals including human beings causes permanent hearing loss because the cochlea cannot regenerate hair cells spontaneously. Here we show that the newborn mouse cochleae contain sphere-forming cells that have the capacity for proliferation in culture, differentiating to form cells that express hair cell markers. When treated with epidermal growth factor or basic fibroblast growth factor, the number of spheres formed increases. The sphere cells express genes that are indicative of inner ear progenitor cells. After differentiation, some sphere cells grow a hair cell bundle-like structure that expresses hair cell marker myosin VIIA and espin. The sphere-forming cells being capable of differentiating into hair cell-like cells implies the possibility of using these sphere-forming cells for reconstructing the damaged cochlear hair cells. PMID- 16708013 TI - Expression of the dopamine transporter in rat and bullfrog retinas. AB - We examined the expression of the dopamine transporter in rat and bullfrog retinas by immunohistochemistry. In both species, the dopamine transporter was strongly expressed in somata and processes of all dopaminergic amacrine cells. In contrast, no immunoreactivity for dopamine transporter was observed in cholinergic amacrine cells. In rat dopaminergic interplexiform cells, dopamine transporter immunoreactivity was also observed on the ascending processes terminating in the outer plexiform layer. Furthermore, the labeling for dopamine transporter diffusely appeared in both outer and inner plexiform layers. This expression profile of the dopamine transporter suggests that dopamine may be taken up not only in the synapses but also extrasynaptically by dopamine transporter, diffusely distributed in both plexiform layers. PMID- 16708014 TI - Neonatal exposure to MK801 induces structural reorganization of the central nervous system. AB - Schizophrenia, a progressive disorder displaying widespread pathological changes, is associated with the loss of glutamatergic function and selective loss of cytoskeletal proteins, such as MAP2, in regions severely affected by this disease. As schizophrenia is associated with perinatal brain trauma, we monitored changes in several functionally different proteins following injury-promoting MK801 blockade of N-methyl-D-aspartate receptors in neonatal rats. Within the somatosensory cortex, MK801 triggered robust, caspase-3-dependent apoptotic injury, reduced expression of cytoskeletal proteins MAP2 and tau, and increased synapse associated protein SNAP25. Thus, both neuronal injury and loss of structural elements important for successful cell-cell contact may reorganize brain circuitry, which at later ages could promote similar behavioral changes observed in schizophrenia. PMID- 16708015 TI - Impaired detection of silent interval change in schizophrenia. AB - The reliably observed reduction in the mismatch negativity response to a change in sound duration in schizophrenia patients may be related to more widespread temporal processing impairments associated with the disorder. This study explored whether individuals with schizophrenia would show electrophysiological evidence of automatic deficits in the ability to detect changes in a regularly repeating interval between two tones. The established pattern was a repeating sequence of a pair of 50-ms tones with a 50-ms silent interval between them. In 8% of pairs, the silent interval between tones was increased to 125 ms. Significant differences in the response to the deviant tone pair in the patient group were consistent with impaired representation of the temporal relationship between sounds. PMID- 16708016 TI - Auditory enhancement of visual temporal order judgment. AB - Although numerous studies have shown that response times can be speeded by the presentation of multisensory stimuli, here we show that such speeding can be seen even when the second sensory channel fails to provide any task-relevant (i.e. redundant) information, and where cueing appears an unlikely explanation. Study participants performed a visual temporal order judgment task in the presence of task uninformative auditory cues, with the latter sound delayed relative to the latter visual cue. Responses were maximally speeded when the auditory stimulus was delayed by a short time (i.e. 100 ms) relative to the second visual target. These results illustrate a unique form of temporal benefit underlying a multisensory interaction, and form the basis for a novel explanation of these perceptual enhancements. PMID- 16708017 TI - Visual field dependence-independence before and after unilateral vestibular loss. AB - To investigate whether visual field dependence-independence changed after unilateral vestibular loss, Meniere's patients were tested before and after unilateral vestibular neurotomy and compared with controls. Using the rod and frame test, visual vertical perception was tested under four visual contexts (with a frame tilted either clockwise or counterclockwise, with a vertical frame, and without visual reference). Both controls and Meniere's patients before unilateral vestibular loss split into visual field dependent and independent subpopulations. Unilateral vestibular loss did not change this partition but, for both subpopulations, it induced asymmetrical visual field dependence with a reduced or abolished contralesional dependence. Finally, when vertical/horizontal references were provided, visual vertical perception was improved in both subpopulations, suggesting that all patients relied on the same allocentric strategy. PMID- 16708018 TI - Asymmetrical facilitation of motor-evoked potentials following motor practice. AB - Use-dependent facilitation of motor-evoked potentials evoked by transcranial magnetic stimulation with repetition of simple movements has been well established. Motor-evoked potentials were recorded from two intrinsic hand muscles before and after blocks of motor practice in which study participants made repeated ballistic pinch responses with either their left or their right hand. Despite similar increases in behavioral performance by each hand (measured by the peak acceleration of the force generated by the index finger), practice related increases in the amplitude of the motor-evoked potentials were greater in the left than in the right motor cortex of right-handed participants. This finding supports the hypothesis that the dominant motor cortex has a greater ability to reorganize with experience than the non-dominant motor cortex. PMID- 16708019 TI - Feature selectivity in area 21a of the cat. AB - We compared the feature tuning of neuronal activity in area 21a with the tuning in areas 17/18. Local field potentials and multi-unit activity recorded in alert animals showed similar selectivity to orientation in both areas, higher selectivity to spatial frequencies in areas 17/18 and higher selectivity and tuning significance to temporal frequencies in areas 17/18. In addition, only at sites in areas 17/18 did the local field potential exhibit locking to a horizontal motion pattern extracted from a natural movie. These results suggest that area 21a is concerned with the analysis of spatial features but lacks a faithful representation of temporal features. Hence, they foster the hypothesis that cortical area 21a is part of a ventral form pathway. PMID- 16708020 TI - GPR39 receptor expression in the mouse brain. AB - GPR39, an orphan G protein-coupled receptor, has been recently identified as the receptor for the bioactive peptide obestatin. Obestatin is secreted from the stomach and acts as an anti-appetite hormone. This activity is induced whether obestatin is administered intraperitoneally or intracerebroventricularly. GPR39 is known to be expressed in the central nervous system but its precise localization is unknown. In view of the growing importance of this system, we decided to study the sites of GPR39 mRNA expression by in-situ hybridization. We find the highest levels of GPR39 mRNA in the amygdala, the hippocampus, and the auditory cortex and low levels in several other brain regions. Surprisingly, we find no expression of GPR39 in the hypothalamus, expected to be the site of the anorexigenic action of obestatin. PMID- 16708022 TI - Caveolin-1 inhibits neurite growth by blocking Rac1/Cdc42 and p21-activated kinase 1 interactions. AB - Growth factors such as basic fibroblast growth factors (bFGFs) could induce the differentiation of mouse neuroblastoma cells. We examine the effect of caveolin-1 on bFGF-induced differentiation of N2a cells. Caveolin-1 blocked the formation of neurites and the phosphorylation of Erk upon bFGF treatment in N2a cells. Active mutants of Rho family small GTPases (Rac1 and Cdc42) could not affect the inhibitory effect of caveolin-1, but we could restore the differentiation of N2a cells by introducing active mutants of p21-activated kinase 1 (PAK1). Over expressed caveolin-1 could be coimmunoprecipitated with PAK1, which interrupted the steady-state Rac1/Cdc42-PAK1 interactions. From these results, we suggest that the up-regulated caveolin-1 in neuronal cells can inhibit the bFGF signaling pathway from small GTPases to PAK1 by directly binding to PAK1. PMID- 16708021 TI - Exposure to a broad range of odorants decreases cell mortality in the olfactory bulb. AB - Experience with multiple odorants during early postnatal development increases the number of cells in the olfactory bulb of rats. In this study, we asked whether at least part of this increase was due to decreased cell death. We selected 30 natural odorants or synthetic odorant mixtures to stimulate a broad area of the bulb during postnatal days 1-15, and counted the number of cells with DNA damage associated with cell death in both the glomerular and the granule cell layers of the main olfactory bulb. Early olfactory enrichment significantly decreased cell death in both bulbar laminae. Thus, olfactory enrichment can spare bulbar cells during early development, possibly leading to increased efficacy in bulb function and enhanced bulbar responses. PMID- 16708024 TI - Space re-exploration in hemispatial neglect. AB - Exploration of the space around us is a fundamental part of human behaviour. When it breaks down there is an important opportunity to understand its underlying mechanisms. Here we show that many right-hemisphere patients with left neglect re explore rightward locations, failing to keep track of them during search. Importantly, such re-exploration occurred despite leftward stimuli being indistinguishable in peripheral vision, so it is unlikely to result from implicit processing of neglected targets. Revisits generally occurred after visits to other targets and are therefore not immediate perseverations. Finally, manipulating the visual salience of found targets altered the degree of neglect, but not revisit rates. Space exploration appears to be modulated both by the ability to keep track of spatial locations and by stimulus salience. PMID- 16708023 TI - Nonprogressive transgene-related callosal and hippocampal changes in PDAPP mice. AB - We have previously shown that homozygous PDAPP mice, a transgenic model of Alzheimer's-like amyloidosis, have abnormal corpus callosi and anterior hippocampi. Now, we investigated the extent to which these morphological abnormalities are correlated with mutant gene dose in a larger, independent, and substantially younger cohort. Homozygous and heterozygous PDAPP mice had significantly smaller callosal commissure length and anterior hippocampal area than controls. Reductions correlated with mutant APP gene dose, with homozygotes showing the greatest reduction, and were present at 2 months of age. These findings and previous work with APP knockouts suggest that PDAPP mice have impaired white matter development due to interference with native murine APP. PMID- 16708025 TI - Hypothermic responses to 8-OH-DPAT in the Ts65Dn mouse model of Down syndrome. AB - Recently, we have demonstrated that potassium channels containing G-protein activated potassium channel 2 (GIRK2) subunits play a significant role in hypothermia induced by several neurotransmitter receptor agonists, including the serotonin (5-HT)1A/5-HT7 receptor agonist 8-OH-DPAT [R-(+)-8-hydroxy-2-(di-n propylamino) tetralin]. The GIRK2 gene is located in human chromosome 21 (its mouse ortholog, Girk2, is in mouse chromosome 16). Down syndrome is produced by the trisomy of chromosome 21. Here, we used quantitative radiotelemetry to investigatehypothermic responses to 8-OH-DPAT in the Down syndrome mouse model Ts65Dn (which carries an extra chromosomal 16 segment containing Girk2). Our results indicate that, in relation to euploid controls, Ts65Dn mice display significantly increased hypothermic responses to 8-OH-DPAT. This finding may be relevant to the understanding of previously reported differences in serotoninergic neurotransmission in persons with Down syndrome. PMID- 16708026 TI - Dysfunction in the neural circuitry of emotional self-regulation in major depressive disorder. AB - An inability to self-regulate negative emotions appears to play a pivotal role in the genesis of major depressive disorder. This inability may be related to a dysfunction of the neural circuitry underlying emotional self-regulation. This functional magnetic resonance imaging study was conducted to test this hypothesis. Depressed individuals and controls were scanned while they attempted to voluntarily down-regulate sad feelings. The degree of difficulty experienced during down-regulation of sadness was higher in depressed individuals. Furthermore, there was greater activation in the right dorsal anterior cingulate cortex, right anterior temporal pole, right amygdala, and right insula in depressed individuals. These results suggest that emotional dysregulation in major depressive disorder is related to a disturbance in the neural circuitry of emotional self-regulation. PMID- 16708027 TI - Prefrontal dopamine D4 receptors are involved in encoding fear extinction. AB - The medial prefrontal cortex is known to be crucially involved in the memory for fear extinction. It has been shown that the dopamine D4 receptor is abundantly localized in the medial prefrontal cortex of rodents and that this receptor subtype is involved in the mediation of fear-related behaviour. In the present study, we tested whether dopamine D4 receptors within the medial prefrontal cortex are involved in the learning and encoding of fear extinction. Infusions of the specific dopamine D4 receptor antagonist L-741,741 into the medial prefrontal cortex immediately before fear extinction did not affect fear extinction learning but attenuated the memory on fear extinction on the following day. These results suggest that medial prefrontal cortex dopamine D4 receptors are involved in the consolidation of fear extinction memory. PMID- 16708028 TI - PDAs in the nursing curriculum: providing data for internal funding. PMID- 16708029 TI - The ins and outs of giving welcome speeches. AB - Many new nursing leaders assuming deanships, assistant deanships, or interim deanships have limited education, experience, or background to prepare them for the job. To assist new deans and those aspiring to be deans, the authors of this department, 2 deans, offer survival tips based on their personal experiences and insights. They address common issues, challenges, and opportunities that face academic executive teams, such as negotiating an executive contract, obtaining faculty lines, building effective work teams, managing difficult employees, and creating nimble organizational structure to respond to changing consumer, healthcare delivery, and community needs. The authors welcome counterpoint discussions with readers. PMID- 16708030 TI - Stress and burnout in nursing faculty. AB - Nursing faculty are at risk for stress and burnout that threaten to worsen the current nursing faculty shortage. The author discusses the importance of preventing nursing faculty burnout and presents specific burnout prevention strategies. PMID- 16708031 TI - Can you do the math? Mathematic competencies of baccalaureate degree nursing students. PMID- 16708032 TI - Teaching compassion: multiple sclerosis and the poetry of Molly Holden. AB - Teaching students about the experiences of those suffering from chronic illnesses like multiple sclerosis is important to the development of compassion. Because information on the psychosocial impact of chronic illness is scarce; however, new learning sources must be investigated. The author explores the use of poetry as a valuable supplement to conventional learning sources and teaching tools. Suggestions for integrating poetry into the nursing curriculum and evaluating its effectiveness are included. PMID- 16708033 TI - An interdisciplinary approach to the needs of non-native-speaking nursing students: conversation circles. AB - When some students with English as an additional language did not respond appropriately in the clinical setting, faculty speculated that it was due to lack of proficiency in the English language. Faculty were uncertain of instructional interventions to assist these students. The authors describe how faculty members explored the learning needs of their student population with English as an additional language and offer practical suggestions to help other faculty members. PMID- 16708035 TI - Try a mock trial. AB - In today s litigious climate, it is important for nurses to understand what constitutes malpractice. This knowledge can help them avoid a lawsuit, a process that can be physically, emotionally, and financially draining. It can also guide their practice so that they provide better care for their clients. The elements of malpractice can be taught through the use of a mock trial. A mock trial can be incorporated into any nursing class and is a valuable educational tool in helping students understand both the legal system and how their practice is viewed. The nurse attorney author discusses a mock trial scenario that can be used to impart information and teach the critical thinking skills necessary for successful nursing practice. PMID- 16708034 TI - Nursing students' attitudes toward technology: a national study. AB - Nursing students' predispositions toward technology may be a factor affecting their use of technology in educational and clinical settings. A national survey was conducted to collect attitudinal measures toward technology and data on technology instruction to assist educators with developing information technology curricula. Outcomes indicate an overall positive attitude toward technology; however, participants reports of formal education in the use of technology applications are low. This shortcoming should be addressed through enhancement of nursing core curriculum. PMID- 16708036 TI - Exploring the concept of caring: novel strategies for a diverse student group. AB - Caring for others and being cared for are part of every person s life. People do not care automatically, but learn to care through the caring example of others and who better to teach about the concept of caring than nurses. The author discusses a nursing elective, Concepts in Caring, that is open to all undergraduates in the university. The concepts of caring, health, illness, human need, and human potential are explored in the course using a variety of teaching strategies. PMID- 16708037 TI - The implementation of a study abroad course for nursing. AB - Study abroad programs can enhance cultural awareness and sensitivity of nursing students and help prepare them to care for a multicultural clientele. The author outlines the efforts at one college to organize and initiate a meaningful study abroad course for nursing students to Merida, Mexico. PMID- 16708038 TI - Measuring critical thinking in graduate education: what do we know? AB - Accrediting and specialty organizations agree that a master's education expands critical thinking skills gained during undergraduate study, yet a review of literature revealed a lack of research measuring critical thinking of students enrolled in postbaccalaureate education in nursing and other fields. Of 35 studies retrieved, 17 were conducted on undergraduate nursing students, with the remainder spread across practicing RNs, and master's level students in physical therapy, business, medicine, and nursing. A critique of this research and recommendations for future study are presented. PMID- 16708039 TI - Development and validation of a derived measure of research utilization by nurses. AB - BACKGROUND: Theoretical models are needed to guide strategies for the implementation of research into clinical practice. To develop and test such models, including analyses of complex theoretical constructs and causal relationships, rich datasets are needed. Working with existing datasets may mean that important variables are lacking. OBJECTIVE: The aim of this study was to derive a nursing research utilization variable and validate it using the Promoting Action on Research Implementation in Health Services (PARIHS) conceptual framework on research implementation. METHODS: This study was based on data from two surveys of registered nurses. The first survey (1996; N = 600) contained robust research utilization variables but few organizational variables. The second (1998; N = 6,526) was rich in organizational variables but contained no research utilization variables. A linear regression model with predictors common to both datasets was used to derive a research utilization variable in the 1998 dataset. To validate these scores, four separate procedures based on the hypothesis of a positive relationship between context and research utilization were completed. Mutually exclusive groups reflecting various levels of context were created to accomplish these procedures. RESULTS: The derived research utilization variable was successfully mapped onto the cases in the 1998 dataset. The derived scores ranged from 0.21 to 21.40, with a mean of 10.85 (SD = 3.23). The mean score per subgroup ranged from 8.28 for the lowest context group to 12.75 for the highest context group. One of the validation procedures showed that significant differences in mean research utilization existed only among four conceptually unique context groups (p < .001). These groups showed a positive incremental relationship in research utilization (p < .001; the better the context, the higher the research utilization score). The validity of the derived variable was supported by using the three remaining validation procedures. DISCUSSION: The successful creation and validation of a derived research utilization variable will enable advanced modeling of the relationships between research utilization and individual and organizational characteristics. The findings also support the construct validity of the context element of the PARIHS theoretical framework. PMID- 16708040 TI - Physical functioning: effect of behavioral intervention for symptoms among individuals with cancer. AB - BACKGROUND: Individuals with cancer receiving chemotherapy suffer deterioration in physical functioning due to symptoms arising from the cancer disease process and its treatment. OBJECTIVES: To determine if age, chronic health conditions (comorbidity), stage of cancer, depressive symptomatology, symptom limitations, sex, and site of cancer moderate the effects of cognitive behavioral intervention on physical function and to determine if symptom limitations mediate the effect of the intervention on physical functioning. METHODS: Two hundred thirty-seven individuals with solid tumor cancer (118 experimental and 119 control group) participated in this 10-contact, 18-week randomized control trial. Cognitive behavioral theory guided the nurse-delivered problem-solving experimental intervention. The control group received conventional care. Interviews occurred at baseline and 10, 20, and 32 weeks. RESULTS: Women with breast cancer had significantly better physical functioning than women with lung cancer. Chronic health conditions, symptom limitation, and depressive symptomatology at baseline were found to moderate the effect of intervention on physical function. Symptom limitation, however, was not found to mediate the effect of intervention on physical functioning. DISCUSSION: The intervention was shown to affect physical function trajectories differently for individuals with different personal and health characteristics. Because poor physical functioning is strongly associated with mortality and poor quality of life, this information may be used by health professionals to target interventions to those who might be most responsive. PMID- 16708041 TI - Cardiovascular risk in Hispanic and non-Hispanic preschoolers. AB - BACKGROUND: Cardiovascular disease (CVD) is the leading cause of death in the United States for both men and women among all racial and ethnic groups. Identifying risk factors early in life can facilitate use of preventive strategies to reduce risk and improve health status across the life span. OBJECTIVE: The aim of this study was to identify modifiable (tobacco smoke exposure, physical inactivity, dietary fat intake, overweight, and high blood pressure [BP]) and nonmodifiable (family history, gender, and age) cardiovascular risk factors in low-income preschool children. METHODS: Low-income preschool children (N = 205) 3-5 years old were recruited to participate. Parents completed a multigenerational cardiovascular health history form and a 24-hour dietary recall for themselves and their child. The children's height, weight, and BP were obtained. RESULTS: Of the 205 children, 61% reported ethnicity as Latino or Hispanic, 31.7% non-Hispanic White, 1% non-Hispanic Black, 3.9% Asian, and 2.4% mixed race. The number of males (50.7%) and females (49.3%) was similar. Only 22 (10.7%) children had no identified cardiovascular risk factors. At least one modifiable risk factor was present in 179 (87.3%) children. Fifty-two (25.5%) children had a body mass index (BMI) > or = 85th percentile for gender and age; 44 (22.3%) had a systolic or diastolic BP over the 90th percentile for gender, age, and height; 128 (66.3%) had a dietary fat intake of > 30%; 77 (37.6%) watched TV or played video games more than 2 hr/day; and 48 (23.4%) were exposed to passive tobacco smoke. DISCUSSION: The identification of cardiovascular risk factors in almost 90% of presumably healthy preschoolers provides evidence to support testing of interventions that can improve health behaviors and reduce risks. PMID- 16708042 TI - Family caregiving in heart failure. AB - BACKGROUND: Little is known about the experiences of family caregivers of patients with heart failure, despite the fact that these patients have disabling symptoms and diminished functioning that could lead to caregiver stress. OBJECTIVES: Based on a caregiver model, the aims of this study were to (a) examine relationships among age, perceived control over managing heart problems, perceived difficulty with tasks, perceived outcomes, and perceived mental and general health among caregivers of persons with heart failure; (b) describe caregivers' perceptions of control over managing heart problems; and (c) describe the tasks and outcomes perceived as being most difficult and negative by caregivers. METHODS: A sample of 21 family caregivers of patients with heart failure completed the study questionnaires. Most were women (n = 20), spouses (n = 20), and White (n = 18), with a mean age of 59.6 years. The patients with heart failure were male veterans with New York Heart Association (NYHA) Class II, III, or IV. Descriptive statistics and Spearman's rho correlations were used. RESULTS: Younger caregivers perceived their tasks to be more difficult and their mental health to be poorer. Caregivers' perceived control over managing heart problems was related moderately to poorer perceived mental health. Greater perceived difficulty with tasks was associated with negative perceptions of caregiver outcomes and poorer perceived mental health. Negative perceptions of caregiver outcomes were associated strongly with perceptions of poorer mental health. Performing household tasks and managing patient behaviors were most difficult, and the caregiver's emotional and financial well-being, time for social activities, and general health had deteriorated. CONCLUSIONS: The preliminary results support the model. Future longitudinal studies are needed in larger samples to evaluate predictors of caregiver-perceived negative outcomes to identify priority areas for interventions. PMID- 16708043 TI - Testing a nurse-tailored HIV medication adherence intervention. AB - BACKGROUND: The relationship between patient adherence and treatment outcomes has been documented across chronic health conditions, but the evidence base for effective adherence interventions in human immunodeficiency virus (HIV) and acquired immune deficiency syndrome (AIDS) requires more rigorous research and reporting. OBJECTIVES: The aims of this study were to determine whether a tailored, nurse-delivered adherence intervention program--Client Adherence Profiling and Intervention Tailoring (CAP-IT)--improved adherence to HIV medications, compared with standard care, and to identify the relationship among adherence measures. METHODS: A randomized controlled trial (RCT) with repeated measures was used to test the efficacy of CAP-IT over a 6-month period. A convenience sample of 240 participants was recruited from a freestanding public HIV/AIDS clinic in Houston, TX, that provides medical, psychological, and pharmaceutical services for over 5,000 clients. Study instruments and measures included demographics; chart audit to capture CD4 count, viral load, and prescribed medications; health literacy; and five measures of adherence (AIDS Clinical Trial Group-Revised Reasons for Missing Medications, Morisky Self-Report of Medication Non-Adherence, Pill Count, Medication Event Monitoring System [MEMS] caps, and Pharmacy Refill). RESULTS: A logistic regression using generalized estimating equations method showed no significant differences over time on the five medication-adherence measures between the experimental and control groups. Little correlation was documented among the five different adherence measures, and there was minimal correlation with clinical markers. DISCUSSION: It is unclear why the tailored adherence intervention was not efficacious in improving medication adherence. The findings suggest that these measures of medication adherence did not perform as expected and that, perhaps, they are not adequate measures of adherence. Effective and efficient adherence interventions are needed to address the barriers to medication adherence in HIV/AIDS. PMID- 16708044 TI - Reducing parenting stress in families with irritable infants. AB - BACKGROUND: Caring for an infant with unexplained, persistent crying is one of the most stressful events for new parents. Infant irritability, also referred to as infantile colic, occurs in 10-25% of all infants and is the most common parental concern reported in the first year of life. OBJECTIVE: The aim of this study was to evaluate the effectiveness of a home-based nursing intervention in reducing parenting stress in three groups of families with irritable infants, using data from a larger evaluation study. METHODS: A two-site, randomized clinical trial was conducted with 121 infants and their parents. Infants were randomized to a treatment or a control group. A third group of infants (n = 43) was added as a posttest-only comparison. The level of parenting stress was measured by the Parenting Stress Index (PSI) at baseline, after the 4-week intervention, and at the 8-week follow-up visit. RESULTS: A repeated measures analysis of variance (ANOVA) was used to compare the two groups across the three time points (baseline, immediately after the 4-week intervention, and at the 8 week follow-up visit). The results revealed a reduction in total parenting stress over time for both the treatment and control groups. Mothers in the treatment group reported reduced parenting stress on the parent-child dysfunctional interaction subscale (p = .04). Total parenting stress scores were found to be significantly higher for mothers in the posttest-only group (p = .009). CONCLUSION: Initial parenting stress levels were high in all participants. Parent feedback at the exit interview indicated that the nurse visits for data collection were also viewed as helpful in reducing the stress level associated with parenting these irritable infants. This home-based program was perceived as helpful in improving the interactions between parents and their irritable infants. PMID- 16708045 TI - Girls on the move program to increase physical activity participation. AB - BACKGROUND: Because physical inactivity poses serious health risks, interventions are urgently needed to reverse the increasingly sedentary lifestyles of adolescent girls. OBJECTIVE: The aim of this study was to determine the feasibility of "Girls on the Move," an individually tailored computerized physical activity (PA) program plus nurse counseling intervention, in increasing PA. METHODS: A pretest-posttest control group design was used with 77 racially diverse sedentary girls in Grades 6, 7, and 8 from two middle schools. Each of the instructional grades was randomly assigned to either an intervention or control condition. After completing computerized questionnaires, each girl in the control group received a handout listing the PA recommendations. To encourage PA, each girl in the intervention group received computerized, individually tailored feedback messages based on her responses to the questionnaires, individual counseling from the school's pediatric nurse practitioner (PNP), and telephone calls and mailings from a trained research assistant. At 12 weeks, girls in both groups responded to the questionnaires. RESULTS: No differences in self-reported PA emerged between the intervention and control groups at Weeks 1 (baseline) and 12 (postintervention). Repeated measures ANOVA showed a significant interaction between group and time for social support for PA, F(1, 69) = 5.73, p = .019, indicating that the intervention group had significantly greater social support across time than did the control group. From baseline to postintervention, social support increased for the intervention group but decreased for the control group. DISCUSSION: Reasons for the lack of significant differences between the groups on the PA measures were cited. Important information that could inform subsequent studies that test interventions to increase youth PA was acquired from conducting this study. Future efforts to increase PA participation might include this approach for enhancing social support for PA. PMID- 16708046 TI - A confirmatory factor analysis of situation-specific Norbeck Social Support Questionnaire items. AB - BACKGROUND: Situation-specific social support, measured by comparably worded Norbeck Social Support Questionnaire (NSSQ) items, holds promise in the development of context-specific theories at the practice level. However, psychometric work on situation-specific NSSQ items is lacking. OBJECTIVE: The aim of this study was to explore the statistical parameters of situation-specific NSSQ items (affect, affirmation, and aid). METHODS: Using data (N = 154) collected for a larger study, the parameters of three LISREL models were estimated by the generally weighted least squares method using an asymptotic covariance matrix. RESULTS: The modified two-factor model was accepted (chi2 = 11.54, df = 7, p < or = .12). Overall, parameter estimates suggest that, in this research context, two situation-specific items (affirmation and aid) measure unique dimensions of social support. CONCLUSIONS: In this instance, situation specific affirmation and aid items measure unique dimensions of support. Guided by strong theory, investigators should develop situation-specific items based on the original NSSQ items. PMID- 16708047 TI - Direct mass spectrometric identification of ABCB1 (P-glycoprotein/MDR1) from the apical membrane fraction of human placenta using fourier transform ion cyclotron mass spectrometry. AB - Identification and quantification of proteins from human tissue by electrophoresis and subsequent mass spectrometry (MS) has been mainly restricted to high abundance, non-membrane-bound molecules. Pharmacologically relevant structures such as cytochrome P450 enzyme, drug transporters and receptors were not accessible by this technology. We developed a method to identify an integral membrane-bound protein (ABCB1) from human placenta using Fourier transform ion cyclotron (FTICR)-MS. Apical and basal membrane fractions were enriched from term human placenta using differential centrifugation. Following SDS-page these fractions were cleaved with trypsin, separated by nano-HPLC and subjected to FTICR-MS. We identified a total of 70 and 89 proteins in the apical and basal membranes, respectively. Among these proteins, and restricted to the apical membrane, was the transport protein ABCB1, with 10 peptides identified by MS, covering a total of 10% of the entire protein. The study describes a method suitable for direct monitoring of membrane-bound proteins from human tissue using FTICR-MS. PMID- 16708048 TI - N-Acetyltransferase and sulfotransferase activity in human prostate: potential for carcinogen activation. AB - BACKGROUND: N-Acetyltransferases (NATs) and sulfotransferases (SULTs) are key phase II metabolizing enzymes that can be involved both in the detoxification and in the activation of many human promutagens and procarcinogens. METHODS AND RESULTS: We investigated the expression of NATs and SULTs in human prostate and tested their role in the activation the N-hydroxy (N-OH) metabolite of 2-amino-3 methylimidazo[4,5-f]quinoline (IQ), a dietary carcinogen, to form DNA adducts. Western blotting showed detectable levels of NAT1, SULT1A1 and SULT1A3 with marked inter-individual variation. NAT2 and other SULT enzymes were not detectable. NAT1 was localized by immunohistochemistry to the cytoplasm of epithelial cells. The presence of acetyl Co-enzyme A (acetyl CoA) and 3' phosphoadenosine-5'-phosphosulfate (PAPS), NAT and SULT cofactors, respectively, significantly increased the level of DNA adducts, detected by P-postlabelling analysis, in calf thymus DNA incubated with N-OH-IQ and prostate cytosolic fractions. The enhancement in the level of DNA adducts in the presence of PAPS correlated with the level of SULT1A1 protein. A single prostate cytosol with the SULT1A1*2/*2 genotype produced less DNA adducts than cytosols with the *1/*2 and *1/*1 genotypes. No significant correlation was observed between NAT1 protein level and the formation of DNA adducts, even in the presence of acetyl CoA. CONCLUSIONS: In conclusion, we demonstrated that NAT1, SULT1A1 and SULT1A3 are present in human prostate and that both enzyme classes significantly contribute to the activation of N-hydroxylated heterocyclic amines to DNA-damaging species in this tissue. Variation in expression levels, in combination with dietary and/or environmental exposure to carcinogens, could be influential in determining individual susceptibility to prostate cancer. PMID- 16708049 TI - Statistical power of association using the extreme discordant phenotype design. AB - BACKGROUND: Selective genotyping has been proven to be an effective design for mapping quantitative trait loci (QTL), either by linkage or by allelic association, wherein the individual trait values can be used as the indices for phenotype selection. It has also been proposed that association studies of dichotomous traits can benefit from such design. When there is no quantitative measurement for phenotype available, cases and/or controls having extreme discordant phenotypes (EDP) can still be selected, based on their exposure status to a drug toxicity or environmental risk factor. The advantage of EDP design is intuitive and it has been successfully used in a number of studies. METHODS: In this report, we developed a statistical method to calculate the power of EDP methodology, using a mixture model of genotype-specific distributions of a single biallelic susceptibility locus. We also compared the power of three statistical tests commonly used in association studies - including the chi test of allelic frequencies, the chi test of genotype frequencies, and the Armitage trend test. The power of two different EDP designs was evaluated under a range of scenarios. RESULTS AND CONCLUSION: Our results indicate that the chi test of genotype frequency is a robust, though less powerful, test for single-locus association, and that EDP methodology is a powerful design for genetic association studies - especially those of common diseases caused by quantifiable drug toxicity or environmental risk factors. PMID- 16708050 TI - Increased CYP3A4 copy number in TONG/HCC cells but not in DNA from other humans. AB - Two recent screens for copy-number variations in the entire human genome found 12.4 gene copy number variations per person, including 2.5% of individuals with gains between 7q21.1 and 7q22.1, the chromosomal location of CYP3A4. CYP3A4 is involved in the metabolism of approximately 50% of all drugs, including many cancer chemotherapeutic agents. CYP3A4 gene copy was determined in DNA from 143 individuals: normal human livers, primary and secondary liver tumors, human hepatic cell lines, and immortalized cell lines representing eight ethnically diverse populations. CYP3A4 gene copy was normal in all but one sample, a primary human hepatocellular carcinoma cell line (TONG/HCC). Southern blots of TONG/HCC DNA revealed an approximate 10-fold increase in CYP3A and a corresponding increase in CYP3A mRNA expression and catalytic activity. Fluorescent in situ hybridization of TONG/HCC revealed specific amplification of the CYP3A4 gene on chromosome 7q21 but no amplification of the MDR1 gene that localizes 11.9 Mb upstream of CYP3A4. High resolution analysis of DNA copy number by comparative genomic hybridization confirmed amplification at 7q21.3-7q22. The amplicon spanned 1.7 Mb and contained 30 known genes, including the entire CYP3A locus. To determine whether CYP3A4 expression affected chemotherapeutic toxicity, LLC-PK1 cells were transduced with adenoviruses expressing CYP3A4 and P450 reductase. CYP3A4 conferred resistance to taxol, vinblastine and topotecan. These studies demonstrate that CYP3A4 copy number differences do not contribute to the normal variation in CYP3A4 expression. Tumors with increased CYP3A copy number (via amplification or increased chromosome 7q) would be expected to show reduced cytotoxicity to some chemotherapeutic drugs and potentially an increase in the outgrowth of drug resistant tumors. PMID- 16708051 TI - Ribonucleotide reductase M1 (RRM1) 2464G>A polymorphism shows an association with gemcitabine chemosensitivity in cancer cell lines. AB - OBJECTIVES: Significant variability in the efficacy and toxicity of an anticancer drug is observed in cancer patients. Currently, there are no standard tools for prediction of a patient's tumor response or his risk of adverse events to chemotherapy. METHODS: We investigate an association between polymorphisms of gemcitabine metabolism-related genes and its chemosensitivity in vitro using 62 human cancer cell lines of various origins. Polymorphisms of gemcitabine metabolism-related genes of deoxycytidine monophosphate deaminase (DCTD), deoxycytidine kinase (DCK) and ribonucleotide reductase M1 (RRM1) were evaluated using the CEQ8000 Genetic analysis system and GeneDoc software. Chemosensitivity of gemcitabine was expressed as an IC50 using MTT assay. RESULTS: The frequency of the polymorphisms was 21% in DCTD 315T>C, 45.2% in RRM1 1082C>A, 59.7% in RRM1 2455A>G, and 79% in RRM1 2464G>A. When examining the association between these polymorphisms and IC50, only the RRM1 2464G>A showed the tendency to be more chemosensitive to gemcitabine (P=0.011), and haplotypes containing 2464G>A polymorphism also showed the association with chemosensitivity when compared to wild-type RRM1 (G2464G). We could not see the significant differences of mRNA expression level with real-time PCR between cell lines according to G2464A polymorphism. In oligonucleotide microarray 73 GenBank Accession Number (69 genes) were selected which expressed differently between RRM1 wild-type and the G2464A polymorphism. CONCLUSIONS: RRM1 2464G>A polymorphism demonstrated an association with gemcitabine sensitivity, which needs functional studies with co expressing genes and prospective clinical studies for the clinical application as a predictive bio-marker. PMID- 16708052 TI - Exon sequencing and high resolution haplotype analysis of ABC transporter genes implicated in drug resistance. AB - BACKGROUND: The ATP-binding cassette (ABC) proteins are a superfamily of efflux pumps implicated as a mechanism for multidrug resistance in cytotoxic chemotherapy, immunosuppressive therapy, HIV and epilepsy. Genetic variation in P glycoprotein, the product of the ABCB1 gene, is proposed to mediate de novo drug resistance, but associations between polymorphisms in ABCB1 and pharmacoresistance have produced conflicting results. Potential explanations for the inconsistency of results include inadequate characterization of gene structure, variation and linkage disequilibrium (LD) in ABCB1, as well as overlap in substrate specificity between ABCB1 and the various other drug transporters. METHODS AND RESULTS: We undertook a fundamental analysis of gene structure, variation and LD in ABCB1 and four other drug transporter genes implicated in pharmacoresistance: ABCC1, ABCC2, ABCC5 and ABCB4. Manual annotation of the five genes revealed nine shorter alternative transcripts with new untranslated regions and one novel region of coding sequence, demonstrating that on-line annotations are incomplete. Sequencing of exons in 47 Caucasian individuals identified 75 novel single nucleotide polymorphisms (SNPs) previously undescribed in any public database, including 14 new coding sequence SNPs. Genotyping of 502 SNPs in 842 Caucasian individuals across the five genes revealed large blocks of high LD, and low haplotype diversity across all five genes that could be characterized by between 67 and 114 tagging SNPs, depending on the tagging criteria. CONCLUSION: The study illustrates that publicly available data resources on genomic organization of genes and common variation can have important gaps and limitations, and establishes a comprehensive set of tagging SNPs for future association studies in pharmacoresistance. PMID- 16708053 TI - Intracisternal A-particle element in the 3' noncoding region of the mu-opioid receptor gene in CXBK mice: a new genetic mechanism underlying differences in opioid sensitivity. AB - OBJECTIVES: CXBK mice, recombinant inbred mice derived from C57BL/6By and BALB/cBy progenitors, display reduced morphine-induced analgesia. Earlier we reported that CXBK mice expressed a reduced amount of the major transcript, MOR-1 mRNA, of the mu-opioid receptor gene. The CXBK MOR-1 mRNA contains a normal coding region and an abnormally long untranslated region. METHODS AND RESULTS: To identify the nucleotide-sequence difference between the CXBK MOR-1 mRNA and that of the progenitors, we first characterized the 3' untranslated region of the MOR 1 mRNA, which was largely unknown. A 3' rapid amplification of cDNA ends-PCR analysis revealed that the 3' untranslated region of the C57BL/6By MOR-1 mRNA was 10 181 nucleotides transcribed from an exon. Next, we compared the MOR-1 genes in C57BL/6By, CXBK, and BALB/cBy mice, and found a 5293 nucleotide insertion only in CXBK mice. The inserted sequence was a variant of the intracisternal A-particle elements that exist in the mouse genome at approximately 1000 sites. Reverse transcription-PCR analyses revealed that the intracisternal A-particle element was transcribed as a part of the CXBK MOR-1 mRNA. No other differences were found in the MOR-1 mRNA between CXBK and BALB/cBy mice, whereas 100 nucleotides differed between C57BL/6By and CXBK mice aside from the intracisternal A-particle insertion. Finally, CXBK mice displayed reduced morphine responses compared with BALB/cBy mice. CONCLUSIONS: Our data suggest that differences in the MOR-1 3' untranslated region appear to cause the CXBK phenotype. This genetic mechanism underlying the CXBK phenotype may provide good insight into the possible genetic mechanisms underlying individual differences in opioid sensitivity in humans. PMID- 16708054 TI - Predicting subsequent infection in patients attending sexually transmitted disease clinics. AB - OBJECTIVE/GOAL: The objective of this study was to identify characteristics associated with subsequent infection in patients attending a sexually transmitted disease (STD) clinic. STUDY DESIGN: Records were retrospectively reviewed for patients from public STD clinics in 4 cities for 12 months after their initial visit to assess subsequent infection with gonorrhea, chlamydia, mucopurulent cervicitis, nongonococcal urethritis, pelvic inflammatory disease, primary or secondary syphilis, or trichomoniasis. RESULTS: Among 64,463 patients, 33.9% had an initial STD and 7.0% had a subsequent STD. Patients with an initial STD had significantly higher probability of a subsequent STD than patients without (12.0% vs. 4.4%). A subsequent STD was significantly more likely for both sexes for those with an initial STD, who were symptomatic at initial visit, reporting exchange of sex, or under age 20 as well as for men reporting sex with men. CONCLUSIONS: Patients with an initial STD were more likely to return with a subsequent STD. Routinely collected information such as initial diagnosis or age can help identify patients at increased risk of a subsequent STD. PMID- 16708055 TI - Syphilis among female sex workers in southwestern China: potential for HIV transmission. AB - OBJECTIVE: The objective of this study was to investigate the prevalence and risk factors of syphilis infection among female sex workers in a southwestern Chinese city along a drug trafficking route. METHODS: From December 2004 to January 2005, 343 female sex workers (FSWs) were recruited through community outreach and peer referring. Confidential questionnaire interviews were administered to collect information on sexual and other sexually transmitted diseases/HIV risk behaviors. Cervical and blood samples were collected to test for syphilis and HIV antibodies and Neisseria gonorrhoeae infection; Condyloma acuminatum infection was diagnosed clinically. RESULTS: Of 343 FSWs, 15.7% were infected with syphilis. The prevalence of HIV, gonorrhea, and C. acuminatum was 0.6%, 2.0%, and 1.5%, respectively. The subtype of 2 HIV-1 infections was both CRF 07BC. Nearly 10% of FSWs reported using illicit drugs. Consistent use, inconsistent use, and never use of condoms with commercial sex clients in the last month were reported by 53.9%, 38.5%, and 7.6% FSWs, respectively. Longer duration of sex work (odds ratio [OR] = 1.98; 95% confidence interval [CI] = 1.08-3.62), working at low-end establishments (OR = 2.03; 95% CI = 1.10-3.76), and if primary sex partners had sex with other women in the past 6 months (OR = 2.06; 95% CI = 1.08-3.91) were independently associated with syphilis infection. CONCLUSIONS: High prevalence of syphilis and overlapped unprotected commercial sex and drug using behaviors among FSWs along a drug-trafficking route may suggest a potential for rapid spread of HIV from injection drug users to FSWs and then to the general population and underscore the urgency of preventive interventions to break the bridge of FSWs for HIV/sexually transmitted disease spread. PMID- 16708056 TI - Place of death after stroke--United States, 1999-2002. AB - Stroke is the third leading cause of death in the United States. Successful acute stroke intervention depends on early recognition of symptoms, prompt emergency transport, and rapid in-hospital treatment. However, approximately half of stroke decedents die before admission to the hospital. During 1990-1998, the proportion of stroke deaths that occurred in hospitals declined, and the proportion occurring before transport to hospitals increased. This report summarizes trends in the place of death among all stroke decedents, the proportion of stroke deaths occurring before emergency assistance arrives, and characteristics associated with place of death. Among 162,672 persons who died of stroke in 2002, 49.2% died pre-transport, 0.4% were dead on arrival (DOA), 3.3% died in emergency departments (EDs), and 47.0% died after admission to a hospital. Early patient and bystander recognition of stroke symptoms and timely action in calling for emergency assistance might reduce the number and proportion of stroke deaths. In addition, improving timely arrival of emergency care and appropriate treatment of stroke patients can reduce the proportion of pre-transport deaths and serious sequelae that lead to severe disabilities. PMID- 16708057 TI - Physical dating violence among high school students--United States, 2003. AB - Dating violence is defined as physical, sexual, or psychological violence within a dating relationship. In a study of dating violence victimization among students in grades 7-12 during 1994-1995, the 18-month prevalence of victimization from physical and psychological dating violence was estimated at 12% and 20%, respectively. In addition to the risk for injury and death, victims of dating violence are more likely to engage in risky sexual behavior, unhealthy dieting behaviors, substance use, and suicidal ideation/attempts. Dating violence victimization can be a precursor for intimate partner violence (IPV) victimization in adulthood, most notably among women. Among adult women in the United States, an estimated 5.3 million IPV incidents occur each year, resulting in approximately 2 million injuries and 1,300 deaths. By using data from the 2003 Youth Risk Behavior Survey (YRBS), CDC analyzed the prevalence of physical dating violence (PDV) victimization among high school students and its association with five risk behaviors. The results indicated that 8.9% of students (8.9% of males and 8.8% of females) reported PDV victimization during the 12 months preceding the survey and that students reporting PDV victimization were more likely to engage in four of the five risk behaviors (i.e., sexual intercourse, attempted suicide, episodic heavy drinking, and physical fighting). Primary prevention programs are needed to educate high school students about healthy dating relationship behaviors, and secondary prevention programs should address risk behaviors associated with dating violence victimization. PMID- 16708058 TI - Prevention of hepatitis A through active or passive immunization: recommendations of the Advisory Committee on Immunization Practices (ACIP). AB - Routine vaccination of children is an effective way to reduce hepatitis A incidence in the United States. Since licensure of hepatitis A vaccine during 1995-1996, the hepatitis A childhood immunization strategy has been implemented incrementally, starting with the recommendation of the Advisory Committee on Immunization Practices (ACIP) in 1996 to vaccinate children living in communities with the highest disease rates and continuing in 1999 with ACIP's recommendations for vaccination of children living in states, counties, and communities with consistently elevated hepatitis A rates. These updated recommendations represent the final step in the childhood hepatitis A immunization strategy, routine hepatitis A vaccination of children nationwide. Implementation of these recommendations will reinforce existing vaccination programs, extend the benefits associated with hepatitis A vaccination to the rest of the country, and create the foundation for eventual consideration of elimination of indigenous hepatitis A virus transmission. This report updates ACIP's 1999 recommendations concerning the prevention of hepatitis A through immunization (CDC. Prevention of hepatitis A through active or passive immunization: recommendations of the Advisory Committee on Immunization Practices [ACIP]. MMWR 1999:48[No. RR-12]:1-37) and includes 1) new data on the epidemiology of hepatitis A in the era of hepatitis A vaccination of children in selected U.S. areas, 2) results of analyses of the economics of nationwide routine vaccination of children, and 3) recommendations for the routine vaccination of children in the United States. Previous recommendations for vaccination of persons in groups at increased risk for hepatitis A or its adverse consequences and recommendations regarding the use of immune globulin for protection against hepatitis A are unchanged from the 1999 recommendations. PMID- 16708059 TI - Youth tobacco surveillance--United States, 2001-2002. AB - PROBLEM/CONDITION: Cigarette smoking is the leading preventable cause of death in the United States, accounting for approximately 440,000 deaths each year. The prevalence of cigarette smoking nationwide among high school students (grades 9 12) increased during the 1990s, peaking during 1996-1997, and then declined. Approximately 80% of tobacco users initiate use before age 18 years. An estimated 6.4 million children aged <18 years who are living today will die prematurely as adults because they began to smoke cigarettes during adolescence. The annual health-related economic cost associated with tobacco use exceeds 167 billion dollars. Because of these health and economic consequences, CDC has recommended that states establish and maintain comprehensive tobacco-control programs to reduce tobacco use among youth. REPORTING PERIOD: This report covers data collected during January 2001-December 2002. DESCRIPTION OF THE SYSTEM: The National Youth Tobacco Survey (NYTS) and state youth tobacco surveys (YTS) were developed to provide states with data to support the design, implementation, and evaluation of comprehensive tobacco-control programs. NYTS is representative of middle and high school students in the 50 states and the District of Columbia. During spring 2002, a total of 26,149 students in 246 schools completed NYTS questionnaires. Weighted data for the YTS were achieved by 13 states in 2001 and by 20 states in 2002; state sample sizes varied (range: 982-38,934). This report summarizes data from the 2002 NYTS and the 2001 and 2002 YTS. RESULTS AND INTERPRETATION: Findings from the 2002 NYTS indicate that current use of any tobacco product ranged from 13.3% among middle school students to 28.2% among high school students. Cigarette smoking was the most prevalent form of tobacco use, with 9.8% of middle school students and 22.5% of high school students reporting that they currently smoke cigarettes. Cigar smoking was the second most prevalent form of tobacco use, with 6.0% of middle school students and 11.6% of high school students reporting that they currently smoke cigars. Among current cigarette smokers, 41.8% of middle school students and 52.0% of high school students reported that they usually smoke Marlboro cigarettes. Black middle school and high school students who smoke were more likely to smoke Newport cigarettes than any other brand (58.3% and 66.8%, respectively). Among middle school students aged <18 years, 75.9% were not asked to show proof of age when they bought or tried to buy cigarettes, and 63.4% were not refused purchase because of their age. Among high school students aged <18 years, 58.5% were not asked to show proof of age when they bought or tried to buy cigarettes, and 60.6% were not refused purchase because of their age. Nearly half (49.6%) of middle school students and 62.1% of high school students who smoke reported a desire to stop smoking cigarettes, with 55.4% of middle school students and 53.1% of high school students reported having made at least one cessation attempt during the 12 months preceding the survey. Among students who have never smoked cigarettes, 21.3% of middle school students and 22.9% of high school students were susceptible to initiating cigarette smoking in the next year. Exposure to secondhand smoke (i.e., environmental tobacco smoke) was high. During the week before the survey, 1) 88.3% of middle school students and 91.4% of high school students who currently smoke cigarettes and 47.1% of middle school students and 53.3% of high school students who have never smoked cigarettes were in the same room with someone who was smoking cigarettes; 2) 81.7% of middle school students and 83.7% of high school students who currently smoke cigarettes and 31.5% of middle school students and 29.1% of high school students who have never smoked cigarettes rode in a car with someone who was smoking cigarettes; and 3) 71.5% of middle school students and 57.5% of high school students who currently smoke cigarettes and 33.3% of middle school students and 29.9% of high school students who have never smoked cigarettes lived in a home in which someone else smoked cigarettes. Media and advertising influence was also noted, with 58.1% of middle school students and 54.9% of high school students who currently use tobacco and 11.0% of middle school students and 13.7% of high school students who have never used tobacco reporting that they would wear or use an item with a tobacco company name or logo on it. Although 84.6% of middle school students and 91.2% of high school students had seen or heard antismoking commercials on television or radio, 89.9% of middle school students and 91.3% of high school students also had seen actors using tobacco on television or in the movies. PUBLIC HEALTH ACTIONS: Health and education officials use YTS and NYTS data to plan, evaluate, and improve national and state programs to prevent and control youth tobacco use. States can use these data in presentations to their state legislators to demonstrate the need for funding comprehensive tobacco-control programs, including tobacco cessation and prevention programs for youth. PMID- 16708060 TI - High-dose chemotherapy and autologous hematopoietic stem cell transplantation for metastatic breast cancer: a therapy whose time has passed. PMID- 16708061 TI - Reduced-dose rasburicase (recombinant xanthine oxidase) in adult cancer patients with hyperuricemia. AB - Recombinant urate oxidase (rasburicase) lowers uric acid levels rapidly to very low levels at the labeled dose of 0.15-0.2 mg/kg daily for 5 days. Our past experience showed that a lower dose (3 mg) lowered uric acid levels sufficiently in most patients. A retrospective review was conducted to determine the effect of a fixed 3 mg dose of rasburicase in 43 adult patients with cancer undergoing hematopoietic stem cell transplantation or receiving chemotherapy who had elevated or rising uric acid levels (6.4-16.8 mg/dl; median 9.6). Six patients received a second dose of rasburicase (3 mg in four patients and 1.5 mg in two patients) 24 h later. Patients received allopurinol, adequate hydration, as well as other supportive therapy as required. Uric acid levels declined by 6-95% (median 43%) within the first 24 h after rasburicase administration, and levels at 48 h were 9-91% (median 65%) lower than the baseline levels. Serum creatinine changed by < or =10% in 21 patients, increased by >10% in four patients and decreased by >10% in 18 patients. No significant renal dysfunction developed in any of the patients. We conclude that rasburicase is effective in lowering uric acid levels at a fixed dose of 3 mg, which is much lower than the recommended dose. PMID- 16708062 TI - Early lymphocyte recovery predicts longer survival after autologous peripheral blood stem cell transplantation in multiple myeloma. AB - To understand the prognostic value of lymphocyte recovery after autologous peripheral blood stem cell transplantation (APBSCT), we performed a retrospective study of 59 newly diagnosed multiple myeloma (MM) patients who underwent frontline APBSCT. Conditioning regimens were melphalan 100 mg/m(2) for 2 days. Following APBSCT, all patients showed complete or partial response. Median follow up time was 29.57 months and median recovery of absolute lymphocyte count (ALC) > or =1000/mm(3) was 23 days. Univariate analysis revealed that significant predictors of overall survival (OS) included bone marrow (BM) plasma cells < or =40% at diagnosis (P=0.0243) and recovery of ALC > or =1000/mm(3) by day +23 (P=0.0156). Positive predictors for progression-free survival (PFS) were BM plasma cells < or =40% at diagnosis (P=0.0134) and recovery of ALC > or =1000/mm(3) by day +23 (P=0.0243). Absolute neutrophil count > or =1000/mm(3) on day +12 was marginally significant for OS and PFS (P=0.0821 and P=0.1153, respectively). Multivariate analysis showed that ALC > or =1000/mm(3) on day +23 independently predicted OS (P=0.031) and prolonged PFS (P=0.011), and that serum beta2-microglobulin was marginally significant for prolonged OS (P=0.066). In conclusion, ALC recovery was an independent predictor of both OS and PFS in MM. PMID- 16708063 TI - Graft-versus-tumor effects on murine mammary carcinoma in a model of nonmyeloablative haploidentical stem cell transplantation. AB - Despite a slight decrease in mortality over the last decade, breast cancer still remains a leading cause of cancer-related death in women. Although anti-tumor effects have been observed after allogeneic stem cell transplantation (SCT), this treatment is not standard care owing to graft-versus-host disease (GVHD) and scarcity of suitable donors. With the aim of reducing treatment-related mortality and increasing donor availability in clinical situations, we developed a preclinical mouse model that combines nonmyeloablative conditioning with the use of haploidentical donor-recipient pairs. To mimic active disease, CB6F1 mice were inoculated with 5 x 10(4) 4T1 mammary carcinoma cells 10 days before transplantation. Keratinocyte growth factor (KGF) was used as GVHD prophylaxis. Syngeneic (CB6F1) SCT did not cure any of the mice and KGF treatment did not influence tumor development. After transplantation with haploidentical (B6CBAF1) bone marrow and splenocytes, however, tumor outgrowth was reduced and long-term disease-free survival (>3 months) was observed in 9/18 (50%) (P=0.0011) of the animals. We conclude that in a model of murine breast cancer, a graft-versus tumor effect can be induced by a nonmyeloablative haploidentical SCT procedure. PMID- 16708064 TI - Successful mobilization and transplantation of filgrastim mobilized hematopoietic stem cells in sickle cell-hemoglobin C disease. PMID- 16708066 TI - PDA-assisted low glycemic index dietary intervention for type II diabetes: a pilot study. AB - BACKGROUND: Epidemiological and dietary intervention studies suggest that a low glycemic index (GI) diet is beneficial for blood glucose control; however, long term clinical utility of the low GI diet has not been fully investigated. OBJECTIVES: To evaluate the feasibility and efficacy of a nutritionist-delivered low-GI dietary intervention, with the support of a personal digital assistant (PDA), for adult patients with poorly controlled type II diabetes. METHOD: The low-GI intervention consisted of six counseling sessions and the use of a PDA based food database with GI scores for 6 months. Study outcomes included feasibility measures, glycosylated hemoglobin levels (HbA1c), GI and glycemic load (GL) score of self-reported dietary intake, body weight, depression and quality of life (QOL). Measures were obtained at baseline, 3 and 6 months. RESULTS: Of 31 adult patients approached, 15 met study eligibility criteria and were enrolled in the study. Thirteen patients (87%) completed all study assessments. Findings included decreases in average HbA1c (-0.5% P = 0.02), body weight, hip circumference, blood pressure, dietary GI and daily caloric intake. Diabetes impact scores also decreased. All but one participant completed all components of the intervention. There were mixed reports regarding the usefulness of the PDAs; however, participants offered helpful suggestions for further development. CONCLUSIONS: Results of this pilot study support the feasibility of implementing a nutritionist-delivered, PDA-assisted low-GI dietary intervention for patients with poorly controlled type II diabetes. Encouraging initial efficacy data require further testing in the context of a randomized clinical trial. PMID- 16708067 TI - Relations between obesity and hypertension: preliminary data from a cross sectional study in primary schoolchildren: the children study. AB - OBJECTIVE: To record the prevalence of overweight, obesity and hypertension in primary schoolchildren living in one of the poorest regions in Europe and furthermore to identify certain behavioural, lifestyle and physiological parameters associated with body mass index (BMI) and blood pressure (BP). DESIGN: Cross-sectional study. SETTING: Nineteen primary schools within the county of Ioannina, Greece. SUBJECTS AND METHODS: Demographic, dietary and physical activity data as well as anthropometrical, BP and cardiorespiratory fitness measurements were obtained from a representative sample of 312 school children (153 boys and 159 girls) attending 5th grade. Inclusion of subjects in the study was voluntary. One-way analysis of variance and multiple linear regression analysis were mainly applied for the evaluation of the tested hypotheses. RESULTS: The prevalence of overweight and obesity was 29.4 and 11.8% for boys and 39.0 and 7.5% for girls. The prevalence of systolic and diastolic hypertension was 28.1 and 7.8% for boys and 26.4 and 17.0% for girls. Furthermore, BMI and BP (systolic and diastolic) were positively related to frequency of fast food meals (rho: + 0.28, P = 0.033, rho: + 1.09, P = 0.03 and rho: +0.86, P = 0.014, respectively) but negatively to leisure time physical activity (rho: -5.55, P = 0.005, rho: -3.32 x 10(-2), P < 0.001 and rho: -2.08 x 10(-2), P < 0.001, respectively). CONCLUSIONS: The current study revealed an increased prevalence of overweight, obesity and hypertension among schoolchildren in Ioannina, indicating the need for early preventive measures in one of the less privileged regions of Europe. PMID- 16708068 TI - Are there associations between coffee consumption and glucose tolerance in elderly subjects? AB - OBJECTIVE: To describe the associations between coffee consumption and glucose tolerance among elderly subjects. DESIGN: A survey among an unselected non institutionalized elderly population. Diabetes was assessed on the basis of self reports and 2-h oral glucose tolerance test for the subjects on diet treatment or with normal glucose tolerance (NGT). Coffee consumption was assessed by a questionnaire. SETTING: Three municipalities in Northern Finland Subjects: All non-institutionalized subjects with baseline normal glucose tolerance or impaired glucose tolerance (IGT) aged 70 years or over. MAIN OUTCOME MEASURES: Incidence, persistence or impairment of abnormal glucose tolerance (AGT). RESULTS: Forty-two percent of those with NGT and 0-5 cups of coffee daily developed AGT, whereas the corresponding figure was 25% for those who drank more than five cups coffee daily. Fifty-five percent of the subjects with IGT or NGT who drank 0-5 cups coffee daily either persisted in IGT or developed AGT. The corresponding figure was 30% for those drinking more than five cups coffee daily. After adjustment for age, body mass index, cardiovascular disease, smoking, alcohol consumption and physical exercise, among those with NGT, the odds ratio (OR) for developing AGT was 2.3 for those drinking 0-5 cups compared to those drinking more than five cups of coffee daily (95% confidence interval (CI) 0.7-7.2). Correspondingly, low coffee consumption was the most powerful predictor associated with the persistence of IGT or the development of AGT (OR 2.9; 95% CI 1.2-6.9). CONCLUSIONS: Low coffee consumption predicts impairment of AGT or persistence of IGT among elderly subjects. PMID- 16708069 TI - Validity of an alternative anthropometric trait as cardiovascular diseases risk factor: example from individuals with traumatic lower extremity amputation. AB - BACKGROUND: Published studies reveal that individuals with lower extremity amputation are vulnerable to cardiovascular diseases (CVD) because of poor physical activity level. Many cardiovascular risk assessment studies have utilized anthropometric traits (primarily body mass index and waist circumference) as cardiovascular risk factor. However, some studies emphasized the technical limitations of measuring waist circumference for studying cardiovascular risk, and so it is difficult to obtain correct measurement from the individuals with lower extremity amputation. OBJECTIVES: The objectives of the present article are to study the prevalence of CVD risk factors among the individuals with traumatic lower extremity amputation and to test the validity of upper arm circumference (UAC) as an alternative anthropometric measurement for screening the CVD risk condition. SUBJECTS AND SETTING: Anthropometric data and other cardiovascular traits data have been collected from unilateral traumatic lower extremity amputated adult males (n = 85) residing in Calcutta and adjoining areas. RESULTS: Results show higher prevalence of cardiovascular risk factor among individuals with above-knee amputation than below-knee amputation. The receiver operating characteristics curve analysis shows significant ability of upper arm circumference to diagnose cardiovascular risk condition. The cutoff value of UAC > 26.6 cm show maximum sensitivity and specificity for the diagnosis of cardiovascular risk condition. Although, binomial tests for equality of proportion does not show any significant difference, however, agreement statistics reveal better diagnostic ability of cutoff value of UAC than the existing cutoff value of waist circumference. CONCLUSIONS: Therefore, UAC provides a better assessment of cardiovascular risk condition than does waist circumference especially for individuals with lower extremity amputation. SPONSORSHIP: Indian Statistical Institute, Kolkata. PMID- 16708070 TI - Molecular genetics of Rett syndrome: when DNA methylation goes unrecognized. AB - The discovery that Rett syndrome is caused by mutations that affect the methyl CpG-binding protein MeCP2 provided a major breakthrough in understanding this severe neurodevelopmental disorder. Animal models and expression studies have contributed to defining the role of MeCP2 in development, highlighting its contribution to postnatal neuronal morphogenesis and function. Furthermore, in vitro assays and microrray studies have delineated the potential molecular mechanisms of MeCP2 function, and have indicated a role in the transcriptional silencing of specific target genes. As well as unravelling the mechanisms that underlie Rett syndrome, these studies provide more general insights into how DNA methylation patterns are recognized and translated into biological outcomes. PMID- 16708072 TI - Inborn errors of metabolism: the flux from Mendelian to complex diseases. AB - Inborn errors of metabolism are characterized by dysregulation of the metabolic networks that underlie development and homeostasis, and constitute an important and expanding group of genetic disorders in humans. Diagnostic methods that are based on molecular genetic tools have a limited ability to correlate phenotypes with subtle changes in metabolic fluxes. We argue that the direct and dynamic measurement of metabolite flux will facilitate the integration of environmental, genetic and biochemical factors with phenotypic information. Ultimately, this integration will lead to new diagnostic and therapeutic approaches that are focused on the manipulation of these pathways. PMID- 16708071 TI - The quest for genetic determinants of human longevity: challenges and insights. AB - Twin studies show that genetic differences account for about a quarter of the variance in adult human lifespan. Common polymorphisms that have a modest effect on lifespan have been identified in one gene, APOE, providing hope that other genetic determinants can be uncovered. However, although variants with substantial beneficial effects have been proposed to exist and several candidates have been put forward, their effects have yet to be confirmed. Human studies of longevity face numerous theoretical and logistical challenges, as the determinants of lifespan are extraordinarily complex. However, large-scale linkage studies of long-lived families, longitudinal candidate-gene association studies and the development of analytical methods provide the potential for future progress. PMID- 16708073 TI - Chromatin remodelling in mammalian differentiation: lessons from ATP-dependent remodellers. AB - The initiation of cellular differentiation involves alterations in gene expression that depend on chromatin changes, at the level of both higher-order structures and individual genes. Consistent with this, chromatin-remodelling enzymes have key roles in differentiation and development. The functions of ATP dependent chromatin-remodelling enzymes have been studied in several mammalian differentiation pathways, revealing cell-type-specific and gene-specific roles for these proteins that add another layer of precision to the regulation of differentiation. Recent studies have also revealed a role for ATP-dependent remodelling in regulating the balance between proliferation and differentiation, and have uncovered intriguing links between chromatin remodelling and other cellular processes during differentiation, including recombination, genome organization and the cell cycle. PMID- 16708074 TI - Size and topology of exogenous DNA as determinant factors of transgene transcription in mammalian cells. PMID- 16708075 TI - p53: more research and more questions. PMID- 16708076 TI - Prevention of murine experimental autoimmune encephalomyelitis by in vivo expression of a novel recombinant immunotoxin DT390-RANTES. AB - Experimental autoimmune encephalomyelitis (EAE) is a T-cell-mediated autoimmune disease. Chemokine receptor CCR5 has been shown to be essential for the T-cell recruitment to the inflammatory site in EAE. In this study, we assumed that an immunotoxin directed at CCR5+ cells would be able to reduce the disease activity of EAE. A recombinant immunotoxin, DT390-RANTES-SRalpha, was constructed in an eukaryotic cell expression plasmid consisting of regulated on activation normal T cells expressed and secreted (RANTES) as the targeting moiety and DT390 as the toxic moiety. DT390-RANTES was expressed in vitro and was highly toxic to activated mouse T cells with the inhibitory concentration 50 at 0.18 ng/ml. To evaluate whether DT390-RANTES was effective in preventing EAE, C57BL/6 mice were immunized with myelin basic protein, emulsified with complete Freund's adjuvant and were treated by injecting cationic liposome-embedded plasmid DNA into the muscle of hind limbs. Mice treated with DT390-RANTES-SRalpha developed a much milder EAE compared to mice treated with phosphate-buffered saline or the empty plasmid DNA. Much less CCR5+-infiltrating cells were found in the central nervous system in DT390-RANTES-SRalpha-treated mice than in the control mice. This study indicates that recombinant immunotoxin can be expressed in vivo, and targeting CCR5 can attenuate the disease activity of EAE in mice. PMID- 16708077 TI - Three catheter-based strategies for cardiac delivery of therapeutic gelatin microspheres. AB - Gelatin hydrogel microspheres (GHMs) have been reported as novel non-viral vectors for gene or protein delivery (GHM therapy). However, the components of an effective catheter-based delivery strategy for GHM therapy are unknown. We evaluated the effectiveness of three catheter-based strategies for cardiac GHM therapy: (1) antegrade injection (AI) via coronary arteries; (2) retrograde injection (RI) via coronary veins; and (3) direct myocardial injection (DI) via the coronary sinus. AI distributed microspheres homogeneously throughout the target area with 73+/-11% retention. RI scattered microspheres non-homogenously with 22+/-8% retention. DI distributed microspheres in the needle-advanced area with 47+/-14% retention. However, despite high efficiency, AI did not show biological effects of inducing angiogenesis from basic fibroblast growth factor bound to GHMs. Furthermore, focal micro-infarctions, owing to micro-embolism of aggregated GHMs into small coronary arterioles, were detected in the AI group. Conversely, only RI and DI groups displayed increased coronary flow reserve. DI groups also demonstrated increased capillary density. These results suggest that RI and DI are effective for cardiac GHM therapy, while AI appears inappropriate owing to the risk of focal infarctions. PMID- 16708078 TI - Modulation of TNFalpha, a determinant of acute toxicity associated with systemic delivery of first-generation and helper-dependent adenoviral vectors. AB - Understanding the determinants of the host innate immune response to systemic administration of adenoviral (Ad) vectors is critical for clinical gene therapy. Acute toxicity occurs within minutes to hours after vector administration and is characterized by activation of innate immune responses. Our data indicate that in mice, indicators of vector toxicity include elevations of cytokine levels, liver transaminase levels and thrombocytopenia. To discern potential targets for blunting this host response, we evaluated genetic factors in the host response to systemically administered first-generation Ad vectors (FGV) and helper-dependent Ad vectors (HDV) containing beta-galactosidase expression cassettes. A preliminary screen for modulation of vector-induced thrombocytopenia revealed no role for interferon-gamma, mast cells or perforin. However, vector-induced thrombocytopenia and interleukin 6 (IL-6) expression are less evident in tumor necrosis factor alpha (TNFalpha)-deficient mice. Moreover, we also demonstrated that TNFalpha blockade via antibody or huTNFR:Fc pretreatment attenuates both thrombocytopenia (>40% increase in platelet count) and IL-6 expression (>80% reduction) without affecting interleukin 12 , liver enzymes, hematological indices or vector transduction in a murine model. Our data indicate that the use of HDV, in combination with clinically approved TNFalpha immunomodulation, may represent an approach for improving the therapeutic index of Ad gene therapy for human clinical trials. PMID- 16708079 TI - AAV-mediated expression of CNTF promotes long-term survival and regeneration of adult rat retinal ganglion cells. AB - We compared the effects of intravitreal injection of bi-cistronic adeno associated viral (AAV-2) vectors encoding enhanced green fluorescent protein (GFP) and either ciliary neurotrophic factor (CNTF), brain-derived neurotrophic factor (BDNF) or growth-associated protein-43 (GAP43) on adult retinal ganglion cell (RGC) survival and regeneration following (i) optic nerve (ON) crush or (ii) after ON cut and attachment of a peripheral nerve (PN). At 7 weeks after ON crush, quantification of betaIII-tubulin immunostaining revealed that, compared to AAV-GFP controls, RGC survival was not enhanced by AAV-GAP43-GFP but was increased in AAV-CNTF-GFP (mean RGCs/retina: 17 450+/-358 s.e.m.) and AAV-BDNF GFP injected eyes (10 200+/-4064 RGCs/retina). Consistent with increased RGC viability in AAV-CNTF-GFP and AAV-BDNF-GFP injected eyes, these animals possessed many betaIII-tubulin- and GFP-positive fibres proximal to the ON crush. However, only in the AAV-CNTF-GFP group were regenerating RGC axons seen in distal ON (1135+/-367 axons/nerve, 0.5 mm post-crush), some reaching the optic chiasm. RGCs were immunoreactive for CNTF and quantitative RT-PCR revealed a substantial increase in CNTF mRNA expression in retinas transduced with AAV-CNTF-GFP. The combination of AAV-CNTF-GFP transduction of RGCs with autologous PN-ON transplantation resulted in even greater RGC survival and regeneration. At 7 weeks after PN transplantation there were 27 954 (+/-2833) surviving RGCs/retina, about 25% of the adult RGC population. Of these, 13 352 (+/-1868) RGCs/retina were retrogradely labelled after fluorogold injections into PN grafts. In summary, AAV-mediated expression of CNTF promotes long-term survival and regeneration of injured adult RGCs, effects that are substantially enhanced by combining gene and cell-based therapies/interventions. PMID- 16708080 TI - Inhibition of human immunodeficiency virus type 1 by RNA interference using long hairpin RNA. AB - Inhibition of virus replication by means of RNA interference has been reported for several important human pathogens, including human immunodeficiency virus type 1 (HIV-1). RNA interference against these pathogens has been accomplished by introduction of virus-specific synthetic small interfering RNAs (siRNAs) or DNA constructs encoding short-hairpin RNAs (shRNAs). Their use as therapeutic antiviral against HIV-1 is limited, because of the emergence of viral escape mutants. In order to solve this durability problem, we tested DNA constructs encoding virus-specific long-hairpin RNAs (lhRNAs) for their ability to inhibit HIV-1 production. Expression of lhRNAs in mammalian cells may result in the synthesis of many siRNAs targeting different viral sequences, thus providing more potent inhibition and reducing the chance of viral escape. The lhRNA constructs were compared with in vitro diced double-stranded RNA and a DNA construct encoding an effective nef-specific shRNA for their ability to inhibit HIV-1 production in cells. Our results show that DNA constructs encoding virus-specific lhRNAs are capable of inhibiting HIV-1 production in a sequence-specific manner, without inducing the class I interferon genes. PMID- 16708081 TI - Direct adenovirus-mediated IGF-I gene transduction of synovium induces persisting synovial fluid IGF-I ligand elevations. AB - Insulin-like growth factor-I (IGF-I) is one of the most influential growth factors in cartilage repair. Maintenance of adequate IGF-I levels after articular repair procedures is complicated by the short biological half-life of IGF-I in vivo. This study investigated the potential for more prolonged IGF-I delivery through direct adenoviral mediated transduction of synovial tissues in the metacarpophalangeal (MCP) joints of horses. The use of a large animal model provided a structurally similar and metabolically relevant corollary to the human knee. The complete IGF-I coding sequence was packaged into an E1-E3 deleted adenovirus-5 vector under cytomegalovirus promoter control (AdIGF-I), and injected at varying total joint doses to the MCP joints of 14 horses. Direct injection of 20 and 50 x 10(10) AdIGF-I resulted in significant elevations of IGF I in synovial fluid for approximately 21 days. Synovial tissue taken from injected joints at day 35 following injection and compared to tissue taken preinjection from the same joints revealed elevated synoviocyte IGF-I mRNA levels for the highest viral dose by in situ hybridization and real-time PCR techniques. AdIGF-I injections did not result in significant lameness, joint effusion or elevated total protein concentrations in the synovial fluid. Mild mononuclear infiltration of white blood cells was evident in histologic sections of the synovium in the second highest adenoviral IGF-I dose of 20 x 10(10) particles. Cartilage biopsies taken from all injected joints did not reveal any significant changes in proteoglycan levels nor in histological morphology, which included chondrocyte cloning, architecture, cell type or toluidine blue staining, when compared to control joints. Based on these findings, gene transfer of IGF-I to the synovium of joints can result in significant and persistent elevations of IGF I ligand in synovial fluid with minimal detrimental effects. Direct IGF-I gene therapy may offer a simple approach in treating patients with acute cartilage injury. PMID- 16708082 TI - Left ventricular hypertrophy determined by Sokolow-Lyon criteria: a different predictor in women than in men? AB - The clinical usefulness of the Sokolow-Lyon voltage criteria in the assessment of electrocardiographic left ventricular hypertrophy (ECG LVH) is addressed. We prospectively studied 3,338 women and 3,330 men referred with hypertension, with an average follow-up of 11.2 years. The voltage amplitude sum SV1+max (RV5 or RV6) was calculated and ECG LVH was defined as a sum >or=3.5 mV. We adjusted survival for age, treatment status before presentation and a previous myocardial infarction or cerebrovascular accident. The risk of stroke, coronary heart disease (CHD) and cardiovascular disease (CVD) mortality increased significantly for each quantitative 0.1 mV increase in baseline electrocardiogram (ECG) voltage, in women within the range of 1.6-3.9% and in men 1.4-3.0%. After further adjustments for race, body mass index, smoking and systolic blood pressure, increasing voltage independently predicted CVD mortality in both men and women. In women, both increasing voltage and the presence of left ventricular hypertrophy (LVH) were predictors of stroke mortality, whereas in men this risk was attenuated. In men, the adjusted association between increasing voltage and CHD mortality tended to be stronger than in women. The use of different thresholds for the two genders made little difference. For stroke and CHD mortality, the population attributable fractions associated with LVH were 15.2 and 5.4% in women and 12.8 and 8.5% in men, respectively. In conclusion, the greater the baseline ECG voltage sum, the greater the associated CVD mortality risk. Women tended to have a high risk of stroke mortality owing to LVH despite adjustments. PMID- 16708083 TI - Adaptive-optics system with liquid-crystal phase-shift interferometer. AB - We develop an adaptive-optics system based on a Mach-Zehnder radial shearing interferometer with liquid-crystal-device (LCD) phase-shift interferometry (PSI). Using accurate phase calibration and transient nematic driving of the LCD, the developed three-step PSI procedure can be achieved in a time of 5 ms. The proposed Mach-Zehnder radial shearing PSI method reconstructs the phase information using a digital signal processor (DSP). The DSP then computes appropriate control signals to drive a deformable mirror in such a way as to eliminate the wavefront distortion. The current adaptive-optics system is capable of suppressing low-frequency thermal disturbances with a signal-to-noise ratio improvement of more than 20 dB and a steady-state phase error of less than 0.02pi root mean square when the control loop is operated at a frequency of 30 Hz. PMID- 16708084 TI - Demonstration and characterization of a detector for minimally destructive detection of Bose condensed atoms in real time. AB - We present the first detector that is capable of recording high-bandwidth atom number density measurements of a Bose-Einstein condensate, with a readout of the data in real time. It is based on a shot-noise-limited unbalanced Mach-Zehnder interferometer designed for a minimally destructive measurement of the atom column density of a Bose-Einstein condensate. The shot-noise limit is reached by phase modulating the laser in one arm and phase locking the interferometer with a second-color laser. The detector is characterized, and its sensitivity for a fractional change in the column density of a Bose-Einstein condensate is calculated. With this detection system it may be possible to implement feedback to stabilize a Bose-Einstein condensate or an atom laser. PMID- 16708085 TI - Optimization of sawtooth surface-relief gratings: effects of substrate refractive index and polarization. AB - The effect of the refractive index of the substrate together with the incident polarization on the optimization of sawtooth surface-relief gratings (SRGs) is investigated. The global optimum diffraction efficiencies of the -1st forward diffracted order of sawtooth SRGs are 63.3% occurring at n2=1.47 for TE polarization and 73.8% occurring at n2=2.88 for TM polarization. Incident TE polarization has higher optimum diffraction efficiency than TM polarization for all n2<1.85. In contrast, TM polarization has higher optimum diffraction efficiency than TE polarization for all n2>1.85. A polymer (n2=1.5) optimum sawtooth SRG exhibits 62.6% efficiency for TE polarization. A silicon (n2=3.475) optimum sawtooth SRG exhibits 68.6% efficiency for TM polarization. These sawtooth SRGs are compared to right-angle-face trapezoidal SRGs. It is found that the optimum profiles of right-angle-face trapezoidal SRGs have only very slightly increased efficiencies over sawtooth SRGs (0.04% for TE and 0.55% for TM). PMID- 16708086 TI - Optical traps with geometric aberrations. AB - We assess the influence of geometric aberrations on the in-plane performance of optical traps by studying the dynamics of trapped colloidal spheres in deliberately distorted holographic optical tweezers. The lateral stiffness of the traps turns out to be insensitive to moderate amounts of coma, astigmatism, and spherical aberration. Moreover holographic aberration correction enables us to compensate inherent shortcomings in the optical train, thereby adaptively improving its performance. We also demonstrate the effects of geometric aberrations on the intensity profiles of optical vortices, whose readily measured deformations suggest a method for rapidly estimating and correcting geometric aberrations in holographic trapping systems. PMID- 16708087 TI - Zoom lens design for a novel imaging spectrometer that controls spatial and spectral resolution individually. AB - A novel imaging spectrometer can individually control spatial and spectral resolution by using zoom lenses as the foreoptics of the system and a focusing lens. By varying the focal length we can use the focusing lens to change the spatial and spectral dimensions; with the foreoptics, however, we can change only the spatial dimension. Therefore the spectral resolution and the spectral range are affected by the zoom ratio of the focusing lens, whereas the spatial resolution and the field of view are affected by the multiplication of the zoom ratios of the foreoptics and the focusing lens. By properly combining two zoom ratios, we can control the spectral resolution with a fixed spatial resolution or the spatial resolution with a fixed spectral resolution. For an imaging spectrometer with this novel zooming function, we used the lens module method and third-order aberration theory to design an initial four-group zoom system with an external entrance pupil for the focusing lens. Furthermore, using the optical design software CODE V, we obtained an optimized zoom lens with a focal-length range of 50 to 150 mm. Finally, the zoom system with its transmission grating in the Littrow configuration performs satisfactorily as the focusing lens of an imaging spectrometer in the wavelength range 450-900 nm. PMID- 16708088 TI - Full-aperture wavefront reconstruction from annular subaperture interferometric data by use of Zernike annular polynomials and a matrix method for testing large aspheric surfaces. AB - We propose a more accurate and efficient reconstruction method used in testing large aspheric surfaces with annular subaperture interferometry. By the introduction of the Zernike annular polynomials that are orthogonal over the annular region, the method proposed here eliminates the coupling problem in the earlier reconstruction algorithm based on Zernike circle polynomials. Because of the complexity of recurrence definition of Zernike annular polynomials, a general symbol representation of that in a computing program is established. The program implementation for the method is provided in detail. The performance of the reconstruction algorithm is evaluated in some pertinent cases, such as different random noise levels, different subaperture configurations, and misalignments. PMID- 16708089 TI - High-speed digital holographic interferometry for vibration measurement. AB - A system based on digital holographic interferometry for the measurement of vibrations is presented. A high-power continuous laser (10 W) and a high-speed CCD camera are used. Hundreds of holograms of an object that has been subjected to dynamic deformation are recorded. The acquisition speed and the time of exposure of the detector are determined by the vibration frequency. Two methods are presented for triggering the camera in order to acquire at a given phase of the vibration. The phase of the wavefront is calculated from the recorded holograms by use of a two-dimensional digital Fourier-transform method. The deformation of the object is obtained from the phase. By combination of the deformations recorded at different times it is possible to reconstruct the vibration of the object. PMID- 16708090 TI - Polarization phase-shifting point-diffraction interferometer. AB - A new instrument, the polarization phase-shifting point-diffraction interferometer, has been developed by use of a birefringent pinhole plate. The interferometer uses polarization to separate the test and reference beams, interfering what begin as orthogonal polarization states. The instrument is compact, simple to align, and vibration insensitive and can phase shift without moving parts or separate reference optics. The theory of the interferometer is presented, along with properties and fabrication techniques for the birefringent pinhole plate and a new model used to determine the quality of the reference wavefront from the pinhole as a function of pinhole size and test optic aberrations. The performance of the interferometer is also presented, along with a detailed error analysis and experimental results. PMID- 16708091 TI - Water jet: a promising method for cutting optical glass. AB - We present an alternative method for cutting optical glass. It works with a high pressure fluid, carring abrasive powder. This technique offers some advantages over conventional methods that use diamond abrasive covered wires or disks. We make a critical comparison between those two techniques, characterizing cuts with interferometric, polarimetric, and Ronchi testing. The main feature of the water jet technique is that it allows surface of any shape, already polished, to be cut safely. PMID- 16708092 TI - Polymer-dispersed liquid-crystal voltage sensor. AB - We present a novel electric-field and voltage sensor based on the electro-optical properties of polymer-dispersed liquid-crystals (PDLCs). In principle, the transmittance of PDLCs is a nonlinear function of the applied electrical field. To measure an AC field we superposed to it a known DC field. This allowed us to achieve linearization of the PDLC response and to measure transmittance changes independently of the light-intensity level variations. Validation experiments are presented. PMID- 16708093 TI - Liquid-crystal panel with microdots on an electrode used to modulate optical phase profiles. AB - The optical characteristics of a liquid-crystal (LC) panel with microdots on an electrode are investigated. Although 3 mum is larger than 1 molecule of LC material, microdots with a 3 microm diameter are sufficiently small to produce a smooth index profile. We use an electrode patterned in a new way to modulate the index profile of the LC panel, which allows us to modulate the optical phase of the passing light. PMID- 16708094 TI - Ferrofluidic adaptive mirrors. AB - A magnetic liquid mirror based on ferrofluids was demonstrated. Magnetic liquid mirrors represent a major departure from solid mirror technology. They present both advantages and disadvantages with respect to established technologies. Stroke (from a fraction of a wave to several hundreds of micrometers), cost (a few dollars per actuator), and scalability (hundreds of thousands of actuators) are the main advantages. Very large mirrors having diameters of the order of a meter should be feasible. There are a few disadvantages. The most important disadvantage is the time response, which is of the order of a few milliseconds. Although this time response could be further decreased with additional technical developments, it is unlikely to match the speed of solid mirrors. The technology is still in its infancy, and considerable work must still be done. However, the advantages are such that the technology is worth pursuing. PMID- 16708095 TI - Light-scattering efficiency of starch acetate pigments as a function of size and packing density. AB - We study theoretically the light-scattering efficiency of paper coatings made of starch acetate pigments. For the light-scattering code we use a discrete dipole approximation method. The coating layer is assumed to consists of roughly equal sized spherical pigments packed either at a packing density of 50% (large cylindrical slabs) or at 37% or 57% (large spheres). Because the scanning electron microscope images of starch acetate samples show either a particulate or a porous structure, we model the coatings in two complementary ways. The material can be either inside the constituent spheres (particulate case) or outside of those (cheeselike, porous medium). For the packing of our spheres we use either a simulated annealing or a dropping code. We can estimate, among other things, that the ideal sphere diameter is in the range 0.25-0.4 microm. PMID- 16708096 TI - Effect of the working gas of the ion-assisted source on the optical and mechanical properties of SiO2 films deposited by dual ion beam sputtering with Si and SiO2 as the starting materials. AB - Silicon and fused-silica targets are used as the starting materials for depositing silicon oxide (SiO2) films. The SiO2 films are prepared by a dual ion beam sputtering deposition system with a main ion source and an ion-assisted source with different working gases. The films deposited are then examined and compared by using a visible spectrophotometer, a Fourier-transform IR spectrophotometer, an atomic force microscope, and contact angle instruments. A Twyman-Green interferometer is employed to study the film stress by phase-shift interferometry. All the SiO2 films show excellent optical properties with extra low extinction coefficients (below 2x10(-5)) and have no water absorption. When the working gas is O2 for the ion-assisted source, the deposited SiO2 films show good properties in terms of stress and roughness and with a good molecular bonding structure order for both targets. However, SiO2 films deposited from the fused-silica target had a larger contact angle, while those deposited from the silicon target had 2.5 times the deposition rate. PMID- 16708097 TI - Three-color mixing for classifying agricultural products for safety and quality. AB - A three-color mixing application for food safety inspection is presented. It is shown that the chromaticness of the visual signal resulting from the three-color mixing achieved through our device is directly related to the three-band ratio of light intensity at three selected wavebands. An optical visual device using three color mixing to implement the three-band ratio criterion is presented. Inspection through human vision assisted by an optical device that implements the three-band ratio criterion would offer flexibility and significant cost savings as compared to inspection with a multispectral machine vision system that implements the same criterion. Example applications of this optical three-color mixing technique are given for the inspection of chicken carcasses with various diseases and for apples with fecal contamination. With proper selection of the three narrow wavebands, discrimination by chromaticness that has a direct relation with the three-band ratio can work very well. In particular, compared with the previously presented two-color mixing application, the conditions of chicken carcasses were more easily identified using the three-color mixing application. The novel three color mixing technique for visual inspection can be implemented on visual devices for a variety of applications, ranging from target detection to food safety inspection. PMID- 16708098 TI - Fast calculation method for spherical computer-generated holograms. AB - The synthesis of spherical computer-generated holograms is investigated. To deal with the staggering calculation times required to synthesize the hologram, a fast calculation method for approximating the hologram distribution is proposed. In this method, the diffraction integral is approximated as a convolution integral, allowing computation using the fast-Fourier-transform algorithm. The principles of the fast calculation method, the error in the approximation, and results from simulations are presented. PMID- 16708099 TI - Characterizing the propagation path in moderate to strong optical turbulence. AB - In February 2005 a joint atmospheric propagation experiment was conducted between the Australian Defence Science and Technology Organisation and the University of Central Florida. A Gaussian beam was propagated along a horizontal 1500 m path near the ground. Scintillation was measured simultaneously at three receivers of diameters 1, 5, and 13 mm. Scintillation theory combined with a numerical scheme was used to infer the structure constant C2n, the inner scale l0, and the outer scale L0 from the optical measurements. At the same time, C2n measurements were taken by a commercial scintillometer, set up parallel to the optical path. The C2n values from the inferred scheme and the commercial scintillometer predict the same behavior, but the inferred scheme consistently gives slightly smaller C2n values. PMID- 16708101 TI - High cw power using an external cavity for spectral beam combining of diode laser bar emission. AB - In extension to known concepts of wavelength-multiplexing diode laser arrays, a new external cavity is presented. The setup simultaneously improves the beam quality of each single emitter of a standard 25 emitter broad-area stripe laser bar and spectrally superimposes the 25 beams into one. By using this external resonator in an "off-axis" arrangement, beam qualities of Mslow2<14 and Mfast2<3 with optical powers in excess of 10 W in cw operation are obtained. PMID- 16708102 TI - Bright diode laser light source. AB - A simplified multiwavelength prototype of an axially symmetric diode laser device based on stacks made of single emitters has been made, and the performance of the device has been demonstrated experimentally. The results verify that kilowatt level light power can be focused into a circular spot with a 1/e2 diameter of 360 microm, a focal length of 100 mm, and a numerical aperture of 0.24, thus producing an average power density in excess of 10 kW/mm2 and a brightness of 6x10(10) W m-2 sr-1. The experiments also predict that it will be possible to increase these values to more than 60 kW/mm2 and 3x10(11) W m-2 sr-1. PMID- 16708103 TI - Bragg mirror inscription on LiNbO3 waveguides by index microstructuration. AB - Numerous applications in integrated optics, especially those related to multiwavelength telecommunications, require dichroic reflectors for use as narrowband or broadband wavelength-selective filters. Bragg mirrors are excellent candidates for this purpose, and we describe a method of fabricating Bragg grating reflectors in Ti-indiffused Lithium Niobate single-mode waveguides based on holographic masking in association with proton exchange. The holographic setup is employed to record a photolithographic mask directly on the substrate, enabling the inscription of waveguides with both periodic and aperiodic distributed parameters. PMID- 16708104 TI - Algorithm improvement and validation of National Institute for Environmental Studies ozone differential absorption lidar at the Tsukuba Network for Detection of Stratospheric Change complementary station. AB - Recently, a data processing and retrieval algorithm (version 2) for ozone, aerosol, and temperature lidar measurements was developed for an ozone lidar system at the National Institute for Environmental Studies (NIES) in Tsukuba (36 degrees N,140 degrees E), Japan. A method for obtaining the aerosol boundary altitude and the aerosol extinction-to-backscatter ratio in the version 2 algorithm enables a more accurate determination of the vertical profiles of aerosols and a more accurate correction of the systematic errors caused by aerosols in the vertical profile of ozone. Improvements in signal processing are incorporated for the correction of systematic errors such as the signal-induced noise and the dead-time effect. The mean vertical ozone profiles of the NIES ozone lidar were compared with those of the Stratospheric Aerosol and Gas Experiment II (SAGE II); they agreed well within a 5% relative difference in the 20-40 km altitude range and within 10% up to 45 km. The long-term variations in the NIES ozone lidar also showed good coincidence with the ozonesonde and SAGE II at 20, 25, 30, and 35 km. The temperatures retrieved from the NIES ozone lidar and those given by the National Center for Environmental Prediction agreed within 7 K in the 35-50 km range. PMID- 16708105 TI - Effect of bio-optical parameter variability and uncertainties in reflectance measurements on the remote estimation of chlorophyll-a concentration in turbid productive waters: modeling results. AB - Most algorithms for retrieving chlorophyll-a concentration (Chla) from reflectance spectra assume that bio-optical parameters such as the phytoplankton specific absorption coefficient (aPhi*) or the chlorophyll-a fluorescence quantum yield (eta) are constant. Yet there exist experimental data showing large ranges of variability for these quantities. The main objective of this study was to analyze the sensitivity of two Chla algorithms to variations in bio-optical parameters and to uncertainties in reflectance measurements. These algorithms are specifically designed for turbid productive waters and are based on red and near infrared reflectances. By means of simulated data, it is shown that the spectral regions where the algorithms are maximally sensitive to Chla overlap those of maximal sensitivity to variations in the above bio-optical parameters. Thus, to increase the accuracy of Chla retrieval, we suggest using spectral regions where the algorithms are less sensitive to Chla, but also less sensitive to these interferences. aPhi* appeared to be one of the most important sources of error for retrieving Chla. However, when the phytoplankton backscattering coefficient (bb,Phi) dominates the total backscattering, as is likely during algal blooms, variations in the specific bb,Phi may introduce large systematic uncertainties in Chla estimation. Also, uncertainties in reflectance measurements, which are due to incomplete atmospheric correction or reflected skylight removal, seem to affect considerably the accuracy of Chla estimation. Instead, variations in other bio-optical parameters, such as eta or the specific backscattering coefficient of total suspended particles, appear to have minor importance. Suggestions regarding the optimal band locations to be used in the above algorithms are finally provided. PMID- 16708106 TI - In situ optical variability and relationships in the Santa Barbara Channel: implications for remote sensing. AB - Relationships and variability of bio-optical properties in coastal waters are investigated. Optical proxies indicate that these coastal waters are optically complex and highly variable and are categorized as follows: (1) relatively clear and dominated by high index of refraction, biogenic particles, (2) more turbid, consisting of mostly inorganic particles and little phytoplankton, (3) extremely turbid with high concentrations of inorganic particles, and (4) more turbid and dominated by biogenic particles. We present a method, alternative to traditional remote-sensing algorithms, of classifying coastal waters [the Spectral Angle Mapper (SAM)] and utilize the SAM to successfully isolate plume conditions in time series of downwelling irradiance and total absorption coefficient. We conclude with a discussion of the use of the SAM for coastal management operations. PMID- 16708107 TI - Spectral variation of the volume scattering function measured over the full range of scattering angles in a coastal environment. AB - The spectral volume scattering function (VSF) was measured in a coastal environment from 0.6 degrees to 177.3 degrees by use of a recently developed device. The spectral variations of the particulate VSF and phase function (i.e., ratio of the VSF to the scattering coefficient) were examined as a function of the scattering angle. The angular dependency of both VSF and phase- function spectra was highly sensitive to the absorption and to the size distribution of the particles. As a result, the use of spectrally neutral phase functions in radiative-transfer modeling is questioned. PMID- 16708108 TI - Noise-resolution trade-off in projection algorithms for laser diffraction particle sizing. AB - The size distribution of a particle suspension can be inferred from studying the diffracted light pattern produced by a laser beam that passes through the suspension. This involves solving an ill-posed linear system. Two previous versions of projection algorithms were tested by use of computer simulations and experiments. Both algorithms showed limitations in restoring the size distribution, because of either the shape of the size distribution or the presence of noise in the scattered light signal. The generalized projection algorithm presented in this work solves these difficulties by introducing a parameter that allows the user to adjust the noise-resolution trade-off, making it suitable for the analysis of natural suspensions. PMID- 16708110 TI - Scattering of a tightly focused beam by an optically trapped particle. AB - Near-forward scattering of an optically trapped 5-mum-radius polystyrene latex sphere by the trapping beam was examined both theoretically and experimentally. Since the trapping beam is tightly focused, the beam fields superpose and interfere with the scattered fields in the forward hemisphere. The observed light intensity consists of a series of concentric bright and dark fringes centered about the forward-scattering direction. Both the number of fringes and their contrast depend on the position of the trapping beam focal waist with respect to the sphere. The fringes are caused by diffraction that is due to the truncation of the tail of the trapping beam as the beam is transmitted through the sphere. PMID- 16708109 TI - Mirror magneto-optical trap using circularly polarized light-emitting optical fibers. AB - A fiber-optic mirror magneto-optical trap (mirror-MOT) that uses a pair of circularly polarized light-emitting optical fibers as an optical access is demonstrated. The fiber is fabricated so that a length of birefringence fiber, designed to be a quarter wave retarder at both wavelengths of 780 and 852 nm, is attached directly onto a polarization-maintaining normal fiber. The polarization states of light emitted from the fibers are sufficiently circular for the operation of a mirror-MOT with 87Rb atoms. The mirror-MOT is able to capture approximately the same number of atoms obtainable with a conventional mirror-MOT. The technique makes it possible to fabricate a compact MOT apparatus by introducing the optical fibers directly into an ultrahigh-vacuum chamber. PMID- 16708111 TI - Simultaneous temperature and exhaust-gas recirculation-measurements in a homogeneous charge-compression ignition engine by use of pure rotational coherent anti-Stokes Raman spectroscopy. AB - Pure rotational coherent anti-Stokes Raman spectroscopy was used for the simultaneous determination of temperature and exhaust-gas recirculation in a homogeneous charge-compression ignition engine. Measurements were performed in a production-line four-cylinder gasoline engine operated with standard gasoline fuel through small optical line-of-sight accesses. The homogenization process of fresh intake air with recirculated exhaust gas was observed during the compression stroke, and the effect of charge temperature on combustion timing is shown. Single-pulse coherent anti-Stokes Raman spectroscopy spectra could not only be taken in the compression stroke but also during the gas-exchange cycle and after combustion. Consequently, the used method has been shown to be suitable for the investigation of two of the key parameters for self-ignition, namely temperature and charge composition. PMID- 16708112 TI - Method of calibration to correct for cloud-induced wavelength shifts in the Aura satellite's Ozone Monitoring Instrument. AB - The in-flight wavelength calibration for the Ozone Monitoring Instrument is discussed. The observed variability in the wavelength scale is two orders of magnitude larger than caused by temperature changes in the instrument. These wavelength variations are the result of rapid changes in time in the radiance levels during an individual observation in the presence of clouds or snow and ice. We have developed a data processing method to account and correct for these changes. In February 2005 this correction was implemented in the official data processing stream. We explain in detail how and how accurately this method works. Before correction, the error in the wavelength scale can be as much as a few tenths of a pixel; after correction it is mostly less than 1/100th of a pixel, which is the required preflight accuracy. This means that higher-level products such as the total column amounts of ozone, NO2, and SO2 are not significantly affected. It is expected that these wavelength variations will be observed in other hyperspectral Earth observation spectrometers and that the correction mechanism should apply equally well. PMID- 16708113 TI - Effect of washing on identification of Bacillus spores by principal-component analysis of fluorescence data. AB - The fluorescence spectra of Bacillus spores are measured at excitation wavelengths of 280, 310, 340, 370, and 400 nm. When cluster analysis is used with the principal-component analysis, the Bacillus globigii spores can be distinguished from the other species of Bacillus spores (B. cereus, B. popilliae, and B. thuringiensis). To test how robust the identification process is with the fluorescence spectra, the B. globigii is obtained from three separate preparations in different laboratories. Furthermore the fluorescence is measured before and after washing and redrying the B. globigii spores. Using the cluster analysis of the first two or three principal components of the fluorescence spectra, one is able to distinguish B. globigii spores from the other species, independent of preparing or washing the spores. PMID- 16708114 TI - Structure, Expression, and Function of a Novel Intercalated Disc Protein, Xin. AB - Xin was first cloned using differential mRNA display from the developing chicken heart. Chick Xin (cXin) participates in a BMP-Nkx2.5-MEF2C pathway to regulating cardiac morphogenesis. Through subsequent EST database searches and cDNA cloning, two mouse Xin genes, mXinalpha and mXinbeta were identified and cloned. The human homologue of mXinalpha (named Cmya1) was mapped to chromosome 3p21.2-p21.3 by radiation hybrid analysis and recently to 3p22.2 by DNA sequencing, which is near the loci for a dilated cardiomyopathy with conduction defect-2 and arrhythmogenic right ventricular dysplasia-5. The predicted human homologue of mXinbeta (named Cmya3) was mapped to chromosome 2q24.3 by DNA sequencing. Predicted Xin proteins all contain a novel 16-amino acid repeating unit (Xin repeat), a putative DNA binding domain and nuclear localization signal, as well as a proline-rich region. All three Xin genes from chick and mouse have a similar tissue expression profile, which is restricted to striated muscle. The expression of mXinalpha in Nkx2.5 or MEF2C knockout mouse embryos was drastically reduced, suggesting that mXinalpha is a downstream target of the Nkx2.5 and MEF2C transcription factors. On the other hand, the expression of mXin was up-regulated when mice were subjected to pressure overload-induced cardiac hypertrophy. Xin protein co localizes with N-cadherin and beta-catenin throughout mouse embryogenesis and into adulthood. Furthermore, mXinalpha appears to interact directly with beta catenin. The Xin repeats bind to actin filaments and may also organize microfilaments into networks. These results may suggest that Xin acts by integrating adhesion, by organizing actin filament arrangement at the insertion sites, and by regulating Wnt/beta-catenin-and N-cadherin-mediated signaling pathways required for cardiac development and cardiac function. PMID- 16708115 TI - Hereditary and acquired thrombophilic disorders. PMID- 16708116 TI - Thrombin generation by exposure of blood to endotoxin: a simple model to study disseminated intravascular coagulation. AB - Pathologic disseminated intravascular coagulation (PDIC) is a serious complication in sepsis. In an in-vitro system consisting of incubation of fresh citrated blood with lipopolysaccharides (LPS) or glucans and subsequent plasma recalcification plasmatic thrombin was quantified. Five hundred microliters of freshly drawn citrated blood of healthy donors were incubated with up to 800 ng/mL LPS (Escherichia coli) or up to 80 microg/mL Zymosan A (ZyA; Candida albicans) for 30 minutes at room temperature (RT). The samples were centrifuged, and 30 microL plasma were recalcified with 1 volume or less of CaCl(2) (25 micromoles Ca(2+)/mL plasma). After 0 to 12 minutes (37 degrees C), 20 microL 2.5 M arginine, pH 8.6, were added. Thirty microliters 0.9 mM HD-CHG-Ala-Arg-pNA in 2.3 M arginine were added, and the absorbance increase at 405 nm was determined. Fifty microliters plasma were also incubated with 5 microL 250 mM CaCl2 for 5, 10, or 15 minutes (37 degrees C). Fifty microliters 2.5 M arginine stops coagulation, and 50 microL 0.77 mM HD-CHG-Ala-Arg-pNA in 2.3 M arginine starts the thrombin detection. The standard was 1 IU/mL thrombin in 7% human albumin instead of plasma. Arginine was also added in the endotoxin exposure time (EET) or in the plasma coagulation reaction time (CRT). Tissue factor (TF)-antigen and soluble CD14 were determined. LPS at blood concentrations greater than 10 ng/mL or ZyA at greater than 1 microg/mL severalfold enhance thrombin generation, when the respective plasmas are recalcified. After 30 minutes EET at RT, the thrombin activity at 12 minutes CRT generated by the addition of 200 ng/mL LPS or 20 microg/mL ZyA is approximately 200 mIU/mL compared to approximately 20 mIU/mL without addition of endotoxin, or compared to about 7 mIU/mL thrombin at 0 minutes CRT. Arginine added to blood or to plasma inhibits thrombin generation; the inhibitory concentration 50% (IC 50) is approximately 15 mM plasma concentration. Endotoxin incubation of blood increases neither TF nor sCD14. This assay allows the study of the hemostasis alteration in PDIC, particularly in PDIC by sepsis. The thrombin generated by blood plus endotoxin incubation and plasma recalcification suggests that the contact phase of coagulation; e.g., triggered by cell components of (phospholipase-) lysed cells such as monocyte or endothelium DNA or phospholipid-vesicles (microparticles), is of primary pathologic importance in sepsis-PDIC. Arginine at plasma concentrations of 10 to 50 mM might be a new therapeutic for sepsis-PDIC. PMID- 16708117 TI - The role of adiponectin in atherosclerosis and thrombosis. AB - Obesity is a major risk factor for morbidity and mortality from cardiovascular causes. Adiponectin has been identified recently as one of the adipocytokines with important metabolic effects. It can suppress atherogenesis by inhibiting the adherence of monocytes, reducing their phagocytic activity, and suppressing the accumulation of modified lipoproteins in the vascular wall. In addition, as adiponectin decrease endothelial damage and stimulates production of NO from vascular endothelial cells, hypoadiponectinemia may be partially contribute to thrombus formation. PMID- 16708118 TI - Vitamin E and alpha-lipoic acid supplementation increase bleeding tendency via an intrinsic coagulation pathway. AB - Vitamin E and alpha-lipoic acid are potent nutritional antioxidants, and when used together, their antioxidant capabilities are improved as alpha-lipoic acid recycles vitamin E. Supplementation of vitamin E has been shown to prolong platelet aggregation but the effects of vitamin E and alpha-lipoic acid supplementation on bleeding tendency have yet to be reported. Young, male rats consumed either control diet (n=5) or vitamin E and alpha-lipoic acid supplemented diet (n=5) for 14 weeks. Activated partial thromboplastin time (APTT) and prothrombin time (PT) were measured as markers of intrinsic and extrinsic coagulation pathways respectively in addition to lipid peroxidation (malondialdehyde). Supplementation significantly prolonged APTT (23.8+/-1.5 vs 31.4+/-1.2s, p<0.05) compared to the control diet; however, there was no significant difference in PT (27.8+/-1.5 vs 26.6+/-0.9s, p>0.05). While vitamin E was increased (p<0.05), there was no significant difference in plasma levels of malondialdehyde (p>0.05). Dietary supplementation of vitamin E and alpha-lipoic acid increases bleeding tendency via inhibition of the intrinsic coagulation pathway with no change in markers of lipid peroxidation. Such supplementation could benefit patients with cardiovascular disease who exhibit elevated levels of coagulation and oxidative stress. PMID- 16708119 TI - Long-term behavior of endothelial and coagulation markers in Eisenmenger syndrome. AB - The long-term behavior of endothelial markers was studied in patients with Eisenmenger syndrome who were subjected to conventional therapy (no vasodilators) and observed for 18 months. Biochemical markers were analyzed comparatively in patients with class II or III symptoms (group 1, n=10) and patients with class IV symptoms (group 2, n=7). Plasma von Willebrand factor antigen (vWF:Ag), thrombomodulin, tissue-type plasminogen activator (t-PA) and its inhibitor (PAI 1), and D-dimer were determined by immunoenzymatic assay at baseline, and at 6, 12, and 18 months. At baseline, the main clinical difference between groups was a decreased peripheral oxygen saturation in group 2 versus group 1 (77+/-5% and 86+/-4%, respectively, p=0.001). Basal vWF:Ag and t-PA were increased and thrombomodulin was decreased in both groups in comparison with controls (p<0.0001), while D-dimer was increased in group 2 only (p=0.0003). In response to treatment, there was a decrease in vWF:Ag in both groups (19% and 23%, respectively in groups 1 and 2, at 18 months vs. baseline, p<0.0001) and t-PA in group 1 (38% vs. baseline, p=0.0485). Plasma vWF:Ag tended to be higher in group 2 in comparison with group 1 during the whole follow-up. Levels of PAI-1 greater than 38.4 ng/mL (upper 90% limit for normals) and D-dimer greater than 500 ng/mL were detected in individual patients (both groups) during the follow-up period. Thrombomodulin remained decreased in both groups. Thus, severity of symptoms in the Eisenmenger syndrome appears to correlate with low oxygen saturation and higher vWF:Ag levels. Improvement of endothelial dysfunction may occur in response to treatment, although increased risk for thrombosis persists, in view of residual abnormalities. PMID- 16708120 TI - Frequency of abnormal biphasic aPTT clot waveforms in patients with underlying disorders associated with disseminated intravascular coagulation. AB - Abnormal biphasic waveform (BPW) patterns were previously reported when the activated partial thromboplastin time (aPTT) was performed in plasma from patients with disseminated intravascular coagulation (DIC). In this study, the prevalence of the BPW was examined in a cohort of 508 hospitalized patients with elevated fibrinogen degradation products (FDP) levels (>10 microg/mL). The presence of a BPW was automatically flagged by the MDA analyzer when the slope of the precoagulation phase in the waveform exceeded a threshold value of 0.25%T/sec. In our cohort, 76 patients (15%) were diagnosed with overt DIC according to the criteria recently proposed by the International Society of Thrombosis and Haemostasis (ISTH), whereas 96 patients (18.9%) were diagnosed with DIC following the criteria of the Japanese Ministry of Health and Welfare (JMHW). The JMHW and ISTH criteria agreed in 93% of cases (kappa coefficient 0.76). The concordance between both scoring systems was high in patients with infection but low in solid cancer. The BPW appeared in 65 patients (12.8%), with the highest prevalence (23.6%) in patients with infection. The BPW was more prevalent in the subgroup of patients with DIC: 59.2% and 47.9% for DIC diagnosed by ISTH and JMWH scores, respectively. The prevalence of the BPW was particularly high in patients with DIC and infection: 86.4% and 75.0% for DIC diagnosed by ISTH and JMWH scores, respectively. For the total cohort, the presence of the BPW was significantly associated with DIC. Odds ratios were 29.9 and 19.0 for ISTH and JMWH scores, respectively (p<0.0001). The BPW showed a moderate sensitivity (59.2% for the ISTH score; 47.9% for the JMWH score), but a high specificity (95.4% for both scores). Waveform analysis of the aPTT potentially provides a practical tool in risk assessment of critical care patients, in whom development of DIC is known to worsen the prognosis. PMID- 16708121 TI - Does hemophilia protect against atherosclerosis? A case-control study. AB - Whether carriers of hemophilia are protected against the development of atherosclerosis is controversial. In a case-control study, the presence of atherosclerosis was assessed and quantified with echo-color Doppler of all explorable arterial districts in 50 carriers of hemophilia and in 50 age-matched control individuals. All participants submitted to echo-color Doppler of carotid and femoral axis, of brachial arteries, and of the abdominal aorta. The presence and grade of atherosclerotic plaques were assessed, as well as the intima-media thickness (IMT). At least one atherosclerotic plaque was found in six cases (12.0%) versus 15 controls (30.0%); referring to the total number of plaques, 30% of them were evaluated in patients affected by decreased coagulation while 70% in subjects with normal levels of FVIII. In all the examined districts, the mean IMT was significantly lower in patients with hemophilia than in controls. Hemophilia protects against asymptomatic atherosclerosis. PMID- 16708122 TI - Hemorrhagic complications in patients treated with anticoagulant doses of a low molecular weight heparin (enoxaparin) in routine hospital practice. AB - Low molecular weight heparins (LMWHs) are a rapidly growing class of anticoagulant drug. Their efficacy has been demonstrated in several clinical settings where they are rapidly becoming the anticoagulant of choice. Controlled clinical studies in patients with deep vein thrombosis, pulmonary embolism, and unstable angina have documented that the frequency of major hemorrhage is 0.5-4%. The purpose of the study was to determine the frequency of minor and major hemorrhage occurring in patients receiving anticoagulant doses of an LMWH (enoxaparin) during routine clinical practice. A prospective, observational study of consecutive patients receiving enoxaparin 1 mg/kg twice daily for at least 24 hours in five internal medicine wards of a university teaching hospital was performed. Five hundred forty-nine patients were studied. The mean age was 67.5+/ 15.5 years and the mean duration of enoxaparin therapy was 3.8+/-1.5 days. Hemorrhage was documented in a total of 94 patients (17.3%). Major hemorrhage occurred in 14 patients (2.6%), injection-site hemorrhage occurred in 55 patients (10%), and minor hemorrhage (noninjection site) was documented in 25 patients (4.7%). There were two deaths attributed to hemorrhage. Patients with major hemorrhage were older than patients with minor or no hemorrhage (75.5+/-10.4 versus 66.8+/-15.2 years; p=0.03) and occurred in patients receiving enoxaparin for a longer period (5.14+/-3.8 days) than those with minor (4+/-2.5 days) or no hemorrhage (2.9+/-2.1 days). Major hemorrhage was significantly associated with impaired renal function, chronic liver disease, and concomitant treatment with warfarin or a proton pump inhibitor. Enoxaparin used in anticoagulant doses in unselected medical patients is not associated with more major hemorrhagic complications than observed in controlled clinical trials. Major hemorrhage may be more likely in older patients, in patients with chronic liver disease and impaired renal function, in patients receiving prolonged enoxaparin therapy, and in patients receiving warfarin or proton pump inhibitors. PMID- 16708123 TI - Comparison of Pycnogenol and Daflon in treating chronic venous insufficiency: a prospective, controlled study. AB - The aim of this study was to investigate the clinical efficacy of oral Pycnogenol (Horphag Research Ltd., UK) in patients with severe chronic venous insufficiency (CVI) in comparison to the combination of diosmin and hesperidin (Daflon, Servier, France). A group of 86 patients with severe chronic venous insufficiency (CVI), venous hypertension, ankle swelling) and previous history of venous ulcerations received either oral Pycnogenol (capsules) 150 mg or 300 mg daily for 8 weeks or Daflon, 1,000 mg/day. All patients completed the study without dropouts. At the end of the study, microcirculatory results indicated: a progressive decrease of skin flux at rest (RF); a significant decrease in capillary filtration (RAS); an improvement in the symptomatic venous score (ASLS); a reduction in edema; a significant improvement (increase) in pO(2) and a decrease in pCO(2) in the Pycnogenol group. A significant level of improvement was reached after 4 weeks of treatment in most patients (p < .05) of the Pycnogenol group while clinical improvement was significant only in 6 subjects in the Daflon group. The positive effects of treatment with Pycnogenol after 8 weeks were significantly larger in comparison with the Daflon group. In conclusion, this study confirms the fast clinical efficacy of Pycnogenol in patients with chronic venous insufficiency and venous microangiopathy and its superiority considering the evaluated parameters-to the combination of diosmin and hesperidin. PMID- 16708124 TI - Monitoring of plasmin and plasminogen activator activity in blood of patients under fibrinolytic treatment by reteplase. AB - There are no reliable data on plasmin or plasminogen activator (PA) activities in blood of patients receiving fibrinolytic treatment. This is due to continuing in vitro action of PA after blood withdrawal. These artefactual changes of PA or plasmin activities have been prevented by arginine stabilization of blood samples of myocardial infarction patients treated with plasminogen activators. Twelve patients with myocardial infarction were treated with reteplase 2 x 10,000,000 units in bolus application; one patient was treated with 100 mg t-PA in continuous infusion. Blood was immediately stabilized with EDTA and arginine. The plasma was analyzed with newly developed assays for plasmin and PA. Maximal plasmin activities in blood were obtained at 40 to 60 minutes reteplase treatment time (0.1-0.6 U/mL = approximately 0.05-0.3 micromol/L plasmin). The 50% clearance rate for plasmatic Pli was greater than 30 minutes. The plasmatic reteplase concentration peaked at approximately 2,000 U/mL after the first bolus infusion and at approximately 1,500-3,500 U/mL after the second bolus infusion. Reteplase was cleared to 50% within less than 30 minutes, also with great inter individual variation. Arginine stabilization of blood allows reliable determinations of activities of plasmin and PA in blood of patients under fibrinolytic treatment: substantial plasmin activities occur in patients treated by reteplase. Therapeutic thrombolysis might be improved, imitating the physiologic cellular thrombolysis; i.e., polymorphonuclear phagocytes (PMN) that can be activated by singlet oxygen ((1)O(2)). PMN might be superior to PA in selective lysis of pathologic thrombi. PMID- 16708125 TI - Pharmacogenomic effect of cytochrome P450 2C9 polymorphisms in different populations. AB - Presently, warfarin sodium or coumadin is the therapeutic drug of choice for maintenance anticoagualtion therapy. One of several factors underlying the variability in warfarin dose response in the patients receiving this oral anticoagulant is a genetic predisposition, especially the CYP2C9 polymorphisms. The pharmacogenomic effect of CYP2C9 polymorphisms is assessed in different racial populations. A retrospective review was performed with an electronic search engine on this topic to get the data for further meta-analysis. A significant correlation between population ethnicity and gene frequencies was detected in this study. A significant low frequency of CYP2C9 variants among the Asian can be demonstrated. The clinical correlation between CYP2C9 polymorphism and warfarin metabolism was also assessed. Four available case-control reports were selected for this meta-analysis. CYP2C9 variants are strongly associated with low-dose warfarin requirement. In summary, the CYP2C9 variants strongly affect the warfarin dose requirement. This phenomenon is ethnically dependent. Due to the high variant frequency among whites in the West, the investigation for CYP2C9 might be useful for this population, not the Asians, whose variant frequency is very low. PMID- 16708126 TI - Prevalence of prothrombin gene mutation (G-A 20210 A) in general population: a pilot study. AB - The objective of this study was to determine the prevalence of prothrombin gene mutation in a sample population from Pakistan. Two hundred apparently healthy unrelated adults (older than 18 years) were included in the study. The sample population comprised 100 Punjabis (male 50, female 50) and 100 Pathans (male 50, female 50). Patients with a history of previous thromboembolism were excluded from the study. Five milliliters (5 mL) of whole blood was drawn in an EDTA bottle. The DNA was extracted by the standard phenol-chloroform method. The DNA was amplified between exon number 14 and the 3'-untranslated region of the prothrombin gene by a polymerase chain reaction in a thermal cycler. Amplified products were digested overnight with HindIII at 37 degrees C. The digested products were electrophoresed on 6% polyacrylamide gel. The fragments were visualized by silver nitrate staining. A heterozygous wild type and an uncut amplified product were included in the electrophoresis strip for quality control. The wild type of DNA ran as a 350-bp fragment and internal control was cut as 550 and 150-bp fragments. The abnormal prothrombin gene was cut into 350-, 322-, and 28-bp fragments. Only two cases of heterozygous prothrombin gene mutation G-A 20210A were found in the sample studied, giving an overall carrier rate of 01% (95% CI 0.4-2.4%) in the target population. Prothrombin gene mutation is present in our population but at a lower frequency than in the white population. PMID- 16708127 TI - Successful treatment of life-threatening bleeding after cesarean section with recombinant activated factor VII. AB - Hemorrhagic shock developed in a 29-year-old nullipara without coagulopathy after emergency caesarean section. Treatment with uterotonic drugs, prostaglandins, and conservative procedures with transfusion of packed red cells and fresh-frozen plasma failed to control the diffuse vaginal and uterine bleeding. Finally an intravenous bolus injection of 90 microg/kg recombinant activated factor VII (rFVIIa, NovoSeven, Novo Nordisk A/S, Bagsvaerd, Denmark), was given and showed success within 20 minutes after administration, without any side effects. PMID- 16708128 TI - An increase of anticardiolipin antibody in association with stroke and chronic chemical exposure. AB - A chemical worker working with urea-formaldehyde resin hazard for 20 years suffered cerebral ischemia in association with an increase of blood beta2 glycoprotein I-dependent anticardiolipin antibody (aCL)-IgG and IgM isotype, and a prolongation of activated partial thromboplastin time (aPTT). Major histocompatibility complex antigen showed DR4 positivity. On follow-up for over 6 years, aCL-IgG and aPTT decreased to reference range but aCL-IgM was still abnormally high despite a cessation of exposure. This patient highlights the induction of antibody-mediated thrombosis in chronic chemical exposure, especially in an individual with subclinical autoimmune disorder. The role of environment for coagulopathic vascular thrombosis is warranted for investigation. PMID- 16708129 TI - Diagnostic and therapeutic difficulties in type 2A von Willebrand disease: resolution. AB - A patient with type 2A von Willebrand disease and a long history of gastrointestinal (GI) bleeding is presented, in whom no abnormality was found on sequencing the von Willebrand factor gene at the DNA level. Subsequent RNA analysis revealed him to be heterozygous for a T-C substitution at nucleotide 4,883, a mutation previously described and associated with type 2A von Willebrand disease. This illustrates the value of a dual DNA/ RNA approach to genetic investigations of highly polymorphic genes. GI bleeding from angiodysplasia is a feature of von Willebrand disease, particularly type 2A. Proactive management with definitive diagnosis of angiodysplasia and ablative treatment where feasible is recommended to stop bleeding symptoms and minimize exposure to blood products. PMID- 16708130 TI - Maternal hemoglobin and pregnancy outcome. PMID- 16708131 TI - von Willebrand factor antigen latex immunoassays are affected to a different extent by rheumatoid factor. PMID- 16708132 TI - Considerations in assessing the clinical course and severity of rotavirus gastroenteritis. PMID- 16708133 TI - Clinical guidelines for celiac disease in children: what does it mean to the pediatrician/family practitioner? PMID- 16708134 TI - Improving influenza vaccination rates in a pediatric asthma management program by utilization of an electronic medical record. AB - An electronic medical record was implemented between 2000 and 2001, and a function of the electronic medical record was utilized as a tool to facilitate improving influenza vaccination rates among moderate to severe asthmatics in the pediatric population in 2002 and 2003. This objective was attempted in order to increase influenza vaccine compliance in the pediatric population. Influenza vaccination levels in this select population were increased 80% in 2002 to 2003 over baseline 2001 levels. Correspondingly, pediatric asthma admissions declined 50% in 2002 to 2003 from a plateau level between 1999 and 2001. All of these results were statistically significant to 95% confidence levels. PMID- 16708135 TI - Pediatric prescribing practices for clonidine and other pharmacologic agents for children with sleep disturbance. AB - The prescription rates of clonidine have risen dramatically and the extent to which these increases can be attributed to treatment of sleep disturbance is unknown. Surveys were mailed to 800 pediatricians across four geographically diverse states to assess prescribing practices specific to sleep disturbance. Ninety-six percent of the respondents treated sleep disturbance. More than one third of the sample reported using clonidine specifically for sleep disturbance including sleep onset, sleep schedule, nighttime awakening, and early morning awakening problems and parasomnias. Clonidine ranked second only to antihistamines as the most commonly used medication for treating sleep disturbance. PMID- 16708136 TI - Lessons learned from pediatric residents on a community pediatrics rotation. AB - Pediatric residents on block community rotations completed journals and exit interviews regarding their perceptions of the rotation. Three common themes present in residents' responses were identified via qualitative analysis: enlightenment and attitude change, impact of direct participation, and rotation challenges. Advantages and disadvantages to block rotations in community pediatrics, and their relationship to learning child advocacy skills, are discussed. Finally, the use of journals as a tool to document systems-based practice competency is explored. PMID- 16708137 TI - Adherence issues related to the selection of antistaphylococcal or antifungal antibiotic suspensions for children. AB - To evaluate the palatability and cost as variables in the selection of antimicrobial suspensions to treat staphylococcal infections and oral thrush, 31 physicians and health care personnel randomly sampled 7 antistaphylococcal antibiotics and 2 antifungal agents, evaluating them in categories of appearance, smell, texture, taste, and aftertaste. Final scoring was then adjusted for cost. Overall taste (palatability) ranking of antistaphylococcal antibiotics, highest to lowest, was as follows: rifampin, cephalexin, EES, TMP/SMX, clindamycin with FLAVORx, linezolid, and clindamycin without FLAVORx. Fluconazole was superior to nystatin. Cost was considered a major factor if the medication was more than $60.00 for treatment of a hypothetical 2-year-old, 13-kg child. PMID- 16708138 TI - Variability in the management of childhood constipation. AB - To assist primary care providers, the North American Society for Pediatric Gastroenterology, Hepatology, and Nutrition (NASPGHAN) published clinical practice guidelines for management of childhood constipation. A cross-sectional survey of pediatricians from across the United States was conducted to assess pediatricians' constipation management strategies, whether pediatricians are familiar with the NASPGHAN constipation guidelines, and reasons pediatricians refer constipated patients to a pediatric gastroenterologist. Overall, 75% of pediatricians used polyethylene glycol without electrolytes to treat childhood constipation, 8% of pediatricians were aware NASPGHAN had published constipation guidelines, and parental pressure was just one reason pediatricians referred constipated patients to a pediatric gastroenterologist. PMID- 16708139 TI - Extreme hyperbilirubinemia in newborn infants. AB - This study was undertaken to determine the frequency and investigate the etiology of extreme hyperbilirubinemia (total serum bilirubin [TSB]>or=25 mg/dL [428 micromol/L]) in newborns admitted to a neonatal intensive care unit in southern Turkey. The charts of 93 term and near-term infants admitted with TSB levels of 25 mg/dL (428 micromol/L) or greater in the first 30 days after birth were retrospectively reviewed. During the 4.5-year study period, 774 infants were admitted to our unit with neonatal jaundice. Ninety-three (12%) of these infants had TSB levels of 25 mg/dL (428 micromol/L) or greater. The mean TSB level in the 93 cases was 30.1+/-5.7 mg/dL (514.7+/-97.5 micromol/L), and the peak levels ranged from 25.0 to 57.4 mg/dL (428-981.5 micromol/L). Thirty-three (35.5%) of the 93 babies had TSB levels of 30 mg/dL (513 micromol/L) or greater. Eighty-nine of 93 infants were being exclusively breast-fed. Nineteen babies were isoimmunized, 7 were bacteremic, 2 of the 39 babies tested for glucose-6 phosphate dehydrogenase had this enzyme deficiency, and 1 of the 71 infants tested for thyroid function had hypothyroidism. No cause for extreme hyperbilirubinemia was found in 61 (65.6%) cases. PMID- 16708140 TI - Preschool vision screening in pediatric practices. AB - A national sample of pediatricians was surveyed to evaluate preschool vision screening practices (response rate 55%, 377 of 690 eligible respondents). The rate of acuity screening for 3-year-old children was low (35%), but increased for 4- (73%) and 5-year-old children (66%). Few used photoscreening or autorefraction (8%). Common barriers were that screening is too time-consuming and children are uncooperative. Half reported that there should be separate reimbursement for vision screening. Financial incentives may be important for ensuring the delivery of preschool vision screening, as well as adoption of emerging screening technologies if such devices are shown to be effective. PMID- 16708141 TI - Accuracy and response time when clerks are used for telephone triage. PMID- 16708142 TI - Hyperphagia following childhood acute lymphoblastic leukemia: a symptom worth heeding. PMID- 16708143 TI - Fever and petechial rash associated with parvovirus B19 infection. PMID- 16708144 TI - Buried penis in the newborn. PMID- 16708145 TI - Concussion in the athlete: a review. PMID- 16708146 TI - Acute facial asymmetry. PMID- 16708147 TI - Highly sensitive child. PMID- 16708149 TI - John Benjamin Murphy (1857-1916): an American surgical phenomenon. PMID- 16708150 TI - Minimally invasive management of colon cancer. AB - One of the most controversial issues in minimally invasive surgery has been the implementation of laparoscopic techniques for the curative resection of colorectal malignancies. Initial concerns included the potential violation of oncologic principles, the effects of carbon dioxide, and the phenomenon of port site tumor recurrence. Basic science research and large randomized controlled trials are now demonstrating that these fears were unjustified. Long-term outcomes of laparoscopic colon resection compared with open colon resection for malignancy are comparable, and there may even be a survival benefit for a subset of patients who undergo laparoscopic resection. PMID- 16708151 TI - Horseshoe abscesses and fistulas: how are we doing? AB - BACKGROUND: Various surgical treatments exist for horseshoe abscesses and fistulae, including posterior midline sphincterotomy, catheter drainage, cutting and draining setons, and advancement flaps. The aim of this study was to evaluate the long-term results of patients treated for these complex anorectal problems. METHODS: A retrospective review was undertaken of patients with a diagnosis of horseshoe abscess, horseshoe fistula, postanal space abscess, or postanal space fistula from 1990 to 2001. Long-term follow-up was accomplished by telephone questionnaire. RESULTS: Thirty-one patients were identified, of whom 17 (54.8%) had a diagnosis of Crohn disease. The diagnosis at presentation included unilateral (ischiorectal) abscess (32.3%), bilateral horseshoe abscess (51.6%), bilateral horseshoe fistula (9.7%), and postanal space abscess (6.4%). Endoanal ultrasonography was used during the preoperative evaluation in 11 patients (35.5%). After referral to our institution, patients underwent a median of four operations (range, 1 to 9). At a mean follow-up of 49.3 months, 60.7% of patients had either healed perineal disease or were asymptomatic with controlled disease. Patients who had a posterior midline sphincterotomy were more likely to be asymptomatic (P=.047). Patients who had a diagnosis of Crohn disease required more operations than those without Crohn disease (3 vs 1.86, P=.02). Only patients who had a diagnosis of Crohn disease had a stoma at their last follow-up (4 of 17, 23.5% vs 0 of 11, 0%; P=.05). CONCLUSIONS: Patients with horseshoe abscess or fistulae often require multiple operations for treatment but can expect reasonable rates of long-term success in controlling or curing their disease. Those who undergo posterior midline sphincterotomy seem to benefit with higher rates of improved symptoms. Patients with a diagnosis of Crohn disease may fare less well. The role of endoanal ultrasonography in directing therapy remains to be defined. PMID- 16708152 TI - Endoluminal methods for gastrotomy closure in natural orifice transenteric surgery (NOTES). AB - Natural orifice transenteric surgery (NOTES) is a new and rapidly evolving concept for intra-abdominal operations that offers the potential for a revolutionary advance in patient care. Conceptually, operations that currently require an open or laparoscopic approach could be performed without incisions in the abdomen, with their concomitant pain and scarring. A recent consensus statement by a joint group of gastrointestinal surgeons and gastroenterologists has identified several technical and technologic hurdles that would need to be overcome before NOTES becomes a clinical reality. One of the most significant requirements identified is the need for a very secure closure of the gastrotomy site that is required for scope passage and specimen removal. Although a rapidly expanding variety of transgastric procedures has been reported, only a few reports address the basic problem of gastrotomy closure. Availability of a safe and simple gastrotomy closure device, however, will be essential for the widespread adoption of the new field of NOTES. Unless new safe and simple devices for endoscopic gastrotomy closure are available and have proven efficacy, NOTES will remain in the hands of a few specialists at centers of excellence because the risk of complications due to insufficient gastrotomy closure will not be acceptable for the surgeon and gastroenterologist in general practice. In this article, we describe three new devices in development or newly on the market that are targeted to advance the safe endoscopic closure of gastrotomy from NOTES. PMID- 16708153 TI - Unsedated small-caliber upper endoscopy: an emerging diagnostic and therapeutic technology. AB - Although conventional esophagogastroduodenoscopy has become widespread in its applications and availability, it is constrained by the requirement for patient sedation. This requirement contributes to morbidity, time lost from work, and additional resource utilization in personnel and facilities. Small-caliber endoscopy is an emerging technology that enables transnasal evaluation of the upper gastrointestinal tract in a unsedated patient. This procedure can be performed in a wider range of settings, including the clinic setting where a dedicated conscious sedation suite is not available and can be incorporated into the office visit. The applications of small-caliber endoscopy include general diagnostic upper endoscopy, screening and surveillance of Barrett esophagus, and intraoperative diagnostics or postoperative evaluation of the upper gastrointestinal tract. Therapeutic applications include the placement of nasoduodenal feeding tubes, esophageal pH catheters, and impedance catheters. When used in the sedated patient, small-caliber endoscopy can also facilitate esophageal stricture dilation and transnasal placement of a percutaneous endoscopic gastrostomy tube. This review discusses the techniques, equipment, and applications of small-caliber endoscopy of the upper gastrointestinal tract. PMID- 16708155 TI - Classification of hiatal hernias using dynamic three-dimensional reconstruction. AB - Hiatal hernias and paraesophageal hernias are common clinical entities and have a well-known classification system. Multiple modalities have been used to illustrate these hernias, most relying on artists' renderings or two-dimensional radiographic studies. However, surgeons would benefit from a comprehensive graphic representation of hiatal hernias based on current imaging technologies. We have applied polygonal mesh surface modeling techniques to render dynamic three-dimensional computed tomography-based models of the four recognized types of hiatal hernias. The resulting images allow nearly real-time navigation in an intuitive and clinically relevant fashion. This model should clarify and eventually advance the existing classification by applying modern and sophisticated image processing to established concepts. PMID- 16708154 TI - Perioperative immunomodulation with Flt3 kinase ligand or a whole tumor cell vaccine is associated with a reduction in lung metastasis formation after laparotomy in mice. AB - INTRODUCTION: Laparotomy has been associated with temporary postoperative immunosuppression and accelerated tumor growth in experimental models. In a previous murine study, a whole cell vaccine plus the adjuvant monophosphoryl lipid A was shown to be effective in decreasing the number of lung metastases that develop after laparotomy. This study was conducted to assess the impact of the adjuvant fetal liver tyrosine kinase 3 (Flt3) ligand on perioperative tumor growth when used alone or with a tumor cell vaccine. METHODS: An intravenous tumor cell injection lung metastases model was used. Sixty female A/J mice were divided into six equal groups designated (1) anesthesia control (AC), (2) AC with Flt3 ligand (ACFlt3), (3) sham laparotomy (OP), (4) OP with Flt3 ligand (OPFlt3), (5) OP with vaccine (OPVac), and (6) OP with Flt3 ligand and vaccine (OPFlt3Vac). Groups 2, 4, and 6 received daily intraperitoneal injections of Flt3 ligand (10 microg/dose with carrier) for 5 days before and 5 days after surgery. Groups 1 and 3 received similar injections of saline on the same schedule. Groups 5 and 6 were vaccinated with irradiated whole Ta3Ha tumor cells intraperitoneally three times before and twice after surgery. Immediately after surgery, all mice were injected with 10(5) Ta3Ha tumor cells via a tail vein. After 14 days, the mice were sacrificed and their lungs and tracheas were excised en bloc. Specimens were stained and counterstained with India ink and Fekete solution, and surface metastases were counted by a blinded observer. Differences between study groups were determined by analysis of variance. The peritumoral inflammatory cell infiltrate of some Flt3 and control specimens was also assessed. RESULTS: Regarding laparotomy, Flt3 ligand (mean, 1.22 metastases), whole cell vaccine (1.12 metastases), and the combination of these two agents (0.1 metastases) were each effective in significantly decreasing the number of surface lung metastases compared with surgery alone (9.88 metastases, P < .05 for all comparisons). There were no differences between the various treatment groups in regards to number of metastases. Only the combination of Flt3 and the vaccine significantly lowered the incidence of tumors (number of mice with > or =1 tumors). Histologic analysis revealed that the Flt3-treated mice demonstrated increased numbers of antigen presenting cells surrounding the tumors compared with controls. CONCLUSIONS: Perioperative treatment with either Flt3 ligand or a whole cell tumor vaccine significantly reduced the number of lung metastases after laparotomy. The combination of the Flt3 ligand and the vaccine also decreased the incidence of metastases and was the most effective treatment. Further studies regarding perioperative immune modulation in the setting of cancer appear warranted. PMID- 16708156 TI - Surgical training and simulation laboratory at Baystate Medical Center. AB - The work hour restrictions imposed by the surgical residency review committee, without a curriculum change, essentially reduces training programs by 30%. The logical result of this is the risk that a higher percentage of surgeons beginning independent practice will fall below the competence level. We believe that simulation will play a vital role in the curriculum to enhance the clinical environment and make the learning environment better and more efficient. Baystate Medical Center (Springfield, Mass) is an affiliate of Tufts University School of Medicine and provides tertiary medical care for the western portion of the state. Our surgical residency program has developed and maintained a simulation center specifically to augment training of surgical knowledge and manual skills. We are also actively involved in education research and curriculum design locally and on a national level. Our center is rapidly becoming a multidisciplinary environment incorporating other residency and clinical programs within the health system and beyond. We are actively pursuing the development of novel assessment technology that will not only integrate the simulation center with the clinical arena but also link educational and clinical outcomes. This will allow us to automatically tailor the educational environment to the individual needs of the learner as they change over time, as well as look at quality improvement related to our educational and research endeavors. PMID- 16708157 TI - Radio frequency identification systems technology in the surgical setting. AB - Radio frequency identification (RFID) is a technology that will have a profound impact on medicine and the operating room of the future. The purpose of this article is to provide an introduction to this exciting technology and a description of the problems in the perioperative environment that RFID might address to improve safety and increase productivity. Although RFID is still a nascent technology, applications are likely to become much more visible in patient care and treatment areas and will raise questions for practitioners. We also address both the current limitations and what appear to be reasonable near future possibilities. PMID- 16708158 TI - Buschke-Lowenstein tumor successful treatment by surgical excision alone: a case report. AB - Perianal giant condyloma acuminatum is a rare variant of condyloma acuminata and often represents a therapeutic challenge. We report a case successfully treated with surgical excision alone. PMID- 16708159 TI - Innovative surgery's dilemma. PMID- 16708160 TI - A new method of assessing anal sphincter integrity using inverted vectormanometry. AB - PURPOSE: Vectorgraphy as an integrated mapping of radial pressure profiles of the anal canal has been used to attempt identification of pressure-related defects with doubtful reliability since vectorgraphs bear no resemblance to endoanal ultrasound scans at similar levels in the anal canal. This study aimed to devise a technique to enable vectorgraphy to be more representative of sphincter function and integrity. METHODS: Vectormanometry was performed in 50 patients with anorectal disorders using an Arndorfer pneumohydraulic system. "Normal" three-dimensional manometric images of each 0.5 cm of the anal sphincter were computer-generated by plotting anal pressures at rest and during squeeze radially around a central zero axis. The graphs were replotted with zero at the periphery and maximal anal pressure at the center. Both this ("inverted") and "normal" vectorgraphs were compared with endoanal ultrasound images at similar levels, assessing both internal and external anal sphincters. RESULTS: Standard vectormanometry produced excellent pictures of pressures throughout the anal canal; the anatomy however bore no resemblance to the pictures produced by endoanal ultrasound. The inverted vectographs showed a much better correlation with endoanal ultrasound at each 0.5-mm level of the anal canal, for both squeeze pressure graphs and external sphincter correlations and for resting pressure graphs and internal sphincter correlations. CONCLUSIONS: Accurate assessment of sphincter integrity is not possible when interpreting the vectormanometry graphs in the current format; however, inverted vectorgraphy gives good correlations with endoanal ultrasound and provides combined functional (pressure measurement) and anatomic (three-dimensional profile) information regarding the anal canal. PMID- 16708161 TI - Red hot chili pepper and hemorrhoids: the explosion of a myth: results of a prospective, randomized, placebo-controlled, crossover trial. AB - PURPOSE: Spicy foods are appreciated by a large part of the world population but have been blamed for causing hemorrhoids or exacerbating their symptoms, although no epidemiologic studies have been performed supporting this hypothesis. In this double-blind, randomized, placebo-controlled, crossover trial, we have studied the effects of a single dose of red hot chili pepper on the hemorrhoidal symptoms. METHODS: Fifty patients with second-degree and third-degree symptomatic hemorrhoids were randomly assigned to take a capsule containing red hot chili powder or placebo during lunch, scoring five hemorrhoidal symptoms (bleeding, swelling, pain, itching, and burning) on a visual analog scale. After one week, crossover treatment was administered according to the same methodology. Other treatments and foods potentially related with anorectal symptoms were discontinued during the study periods. RESULTS: Patients assigned low scores to their hemorrhoidal symptoms before the study and the scores remained unchanged during the 48 hours after both placebo and chili pepper treatment, the latter showing no statistically significant effects. CONCLUSIONS: There is no scientific evidence that a spicy meal based on red hot chili pepper may worsen hemorrhoidal symptoms and, therefore, there is no reason to prevent these patients from occasionally enjoying a spicy dish if they so wish. PMID- 16708165 TI - Purification and characterization of Thermotoga maritima homoserine transsuccinylase indicates it is a transacetylase. AB - The methionine biosynthetic pathway found in bacteria is controlled at the first step, acylation of the gamma-hydroxyl of homoserine. This reaction is catalyzed by one of two unique enzymes, homoserine transacetylase or homoserine transsuccinylase, which have no amino acid sequence similarity. We cloned, expressed, and purified homoserine transsuccinylase from the thermophilic bacterium Thermotoga maritima. Substrate specificity experiments demonstrated that acetyl-coenzyme A (CoA) is the preferred acyl donor and is used at least 30 fold more efficiently than succinyl-CoA. Steady-state kinetic experiments confirm that the enzyme utilizes a ping-pong kinetic mechanism in which the acetate group of acetyl-CoA is initially transferred to an enzyme nucleophile before subsequent transfer to homoserine. The maximal velocity, V/K (acetyl-CoA) and V/K (homoserine), all exhibited bell-shaped pH curves with apparent pKs of 6.0-6.9 and 8.2-8.8. The enzyme was inactivated by iodoacetamide in a pH-dependent manner, with an apparent pK of 6.3, suggesting the presence of an active-site cysteine residue which forms an acetyl-enzyme thioester intermediate during catalytic turnover, similar to observations with other transsuccinylases. In addition, the enzyme is highly stable at elevated temperatures, maintaining full activity at 70 degrees C. Taken together, these data suggest that the T. maritima enzyme functions biochemically as a transacetylase, despite having the sequence of a transsuccinylase. PMID- 16708166 TI - Microarray detection of a de novo der(X)t(X;11)(q28;p13) in a girl with premature ovarian failure and features of Beckwith-Wiedemann syndrome. AB - We report an 18-year-old girl with premature ovarian failure (POF), tall stature, and urinary incontinence. Chromosome studies including array comparative genomic hybridization showed that she was the carrier of an unbalanced de novo translocation between the X chromosome and chromosome 11, resulting in partial monosomy Xq and partial trisomy 11p. Microsatellite analysis demonstrated that the patient had paternal duplication of 11p13p15.5, which contributed to some of her features consistent with Beckwith-Wiedemann syndrome (BWS). The combined phenotype of BWS and POF suggests that the translocated portion of 11p remains active. PMID- 16708167 TI - The HPA axis and perinatal depression: a hypothesis. AB - Episodes of depression and anxiety are as common during pregnancy as postpartum. Some start in pregnancy and resolve postpartum, others are triggered by parturition and some are maintained throughout. In order to determine any biological basis it is important to delineate these different subtypes. During pregnancy, as well as the rise in plasma oestrogen and progesterone there is a very large increase in plasma corticotropin releasing hormone (CRH), and an increase in cortisol. The latter reaches levels found in Cushing's syndrome and major melancholic depression. Levels of all these hormones drop rapidly on parturition.We here suggest that the symptoms of antenatal and postnatal depression may be different, and linked in part with differences in the function of the hypothalamic pituitary adrenal (HPA) axis. There are two subtypes of major depression, melancholic and atypical, with some differences in symptom profile, and these subtypes are associated with opposite changes in the HPA axis. Antenatal depression may be more melancholic and associated with the raised cortisol of pregnancy, whereas postnatal depression may be more atypical, triggered by cortisol withdrawal and associated with reduced cortisol levels. There is evidence that after delivery some women experience mild bipolar II depression, and others experience post traumatic stress disorder. Both of these are associated with atypical depression. It may also be that some women are genetically predisposed to depression of the melancholic type and some to depression of the atypical type. These women may be more or less vulnerable to depression at the different stages of the perinatal period. PMID- 16708169 TI - Endovascular treatment of intracranial aneurysms with parent vessel reconstruction using balloon and self expandable stents. AB - BACKGROUND AND PURPOSE: To assess the feasibility and results of parent vessel stent reconstruction with balloon expandable and self-expandable stents in the treatment of intracranial aneurysms. METHODS: In a total of 18 aneurysms balloon expandable (group A) and self expandable (group B) stents were used in 9 cases each. Stent implantation alone was used in 3 cases, and additional coil packing in the other 15. RESULTS: Stents were successfully deployed in 8 out of 9 in group A and in 9 out of 9 cases in group B. Nearly complete occlusion was achieved in all but one case. At 3 or 6 months stable occlusion was found in 4 group A and 2 group B patients, progressive thrombosis in 3 cases in both groups, and recanalisation in 1 case in group B. Late follow up at 1-4 years demonstrated one progressive thrombosis one recanalisation and 1 stable occlusion in 3 group A, and 2 stable occlusions in 2 group B. patients. Complications included one aneurysm perforation in group A, one in-stent thrombosis and a distal arterial perforation in group B and one groin hematoma in both groups. CONCLUSION: Stent reconstruction of intracranial arteries harbouring aneurysms is feasible and may result in aneurysm thrombosis without coil packing in some cases. Self expanding stents seem to provide a higher rate of success. Aggressive antiplatelet treatment increases the risk of hemorrhagic complications. PMID- 16708168 TI - Bone tumors at the cranio-cervical junction. Surgical management and results from a series of 41 cases. AB - BACKGROUND: Bone tumors located at the cranio-cervical junction (CCJ)are rare. Tumoral involvement of the neighbouring structures including bone, nerves and vertebral artery and the dynamic aspects of the bone structures raise technical difficulties in the surgical approach. The surgical management includes tumoral resection and stabilization of the CCJ. METHODS: Forty-one patients presenting a bone tumor (26 benign and 15 malignant tumors), excluding chordomas, located at the CCJ (including lower third of the clivus, C1 and C2) were observed over 20 years from 1981 to 2001. Imaging work-up included CT scanner with bone windows sequences and reconstruction in the coronal and sagittal plane; since 1984 most of the patients (N=35) underwent a MRI and angioMR scanning. Vertebral angiography was rarely performed (N=9) and mostly when the diagnosis was doubtful. In some cases the diagnosis was clear but in others, imaging studies showed destructive lesions suggesting a malignancy, which sometimes required a biopsy (N=4). The surgical resection was only performed through a lateral approach. FINDINGS: Complete resection was achieved in 38 cases while in 3 cases a small remnant was left behind. A complementary stabilization procedure was necessary in 18 cases using either bone grafting during the same procedure and through the same approach (N=5) or a craniocervical plating and bone grafting (N=13). No recurrence in the group of benign tumors was seen during an average follow-up of 6 years (from 2 to 11 years). The pre-operative symptoms of pain and neck stiffness, improved or disappeared in most patients. Three patients with lower cranial nerves (N=2) or sphincter disturbances (N=1) remained unchanged. One patient with tetraplegia eventually died. CONCLUSIONS: Various types of bone tumors may be found at the CCJ. Confusion between benign and malignant tumor or pseudo tumors must be avoided, sometimes requiring a biopsy. Surgery using a lateral approach, usually permits the surgeon to achieve a complete resection either preserving the stability of the CCJ whenever intact or associated with a stabilization procedure. PMID- 16708170 TI - NON-activated protein C as post-treatment after spinal cord compression injury in rats. AB - BACKGROUND: Neuroprotective effects of recombinant human activated Protein C (rhAPC) in models of Spinal Cord Injury (SCI) and ischemic stroke have been reported in rodents. To rule out immunogenicity of rhAPC and to possibly maintain the physiological PC/thrombin balance the use of zymogen PC in SCI might be preferable. Although activation of Protein C (PC) has been demonstrated in rats, the efficacy and drug safety of NON activated PC has not been previously tested in experimental SCI. METHODS: Twelve rats were subjected to 40 g compression of the spinal cord at TH11 for 20 minutes and randomly allocated to either the NON activated PC (25 IU/kg) or the Placebo group (saline).Results. 25 IU treatment yielded improved recovery from SCI compared to placebo and the triple fold dose of PC (75 IU/kg) was subsequently tested to detect treatment associated complications (TAC). Treatment was administered as a single shot via the right vena jugularis forty minutes after onset of compression. The observation period was 5 weeks in 25 IU treated and 1 week in the 75 IU treated rats. Improvement of motor function recovery was measured with behaviour tests and electrophysiology. FINDINGS: Single shot treatment with 25 IU/kg of NON activated PC led to improved recovery in terms of behaviour and electrophysiology. TACs neither occurred in the 25 IU nor in the 75 IU group within one week. CONCLUSION: NON activated PC is a potent and safe drug in experimental SCI and should be considered for treatment in neurotrauma. PMID- 16708171 TI - Isolation of neuronal progenitor cells from the adult human neocortex. AB - BACKGROUND: The reliability of harvesting neuronal progenitor cells (NPCs) from the adult human neocortex has not been established, with respect to preparing autologous cell cultures for transplantation in stroke and traumatic brain injured patients. METHOD: Enriched NPC cultures have been generated from nonneurogenic regions of the adult rodent brain by buoyancy-dependent fractionation, but the feasibility of using such a method to isolate NPCs from the adult human cortex has not been reported previously. To determine if a starter population of human adult cortical NPCs could be isolated for in vitro expansion using this method, tissue samples from five patients undergoing cortical resection for either epilepsy or trauma were assayed. FINDINGS: Cultured cells generated from all patients predominately expressed both the NPC marker nestin and neuron-specific beta-tubulin III. The presence of NPCs was verified by in vitro BrdU/beta-tubulin III co-labeling and increasing beta-tubulin expression in differentiating conditions. Despite the formation of aggregates in monolayer culture, cell proliferation as measured by BrdU incorporation was not as prevalent as that reported from rodent cultures generated by this protocol. CONCLUSIONS: NPCs isolated from the adult human neocortex using this method expressed beta-tubulin III in larger percentages than has been previously reported for NPCs isolated using other methods. As such, these data suggest the possibility of culturing dividing neuroblasts from the adult neocortex for further manipulation as transplantable cells. PMID- 16708172 TI - Comparative experimental study of argon plasma and bipolar coagulation techniques. AB - BACKGROUND: Argon plasma coagulation (APC) is based on the principle of ionised argon creating conductive plasma between an activating electrode and tissue surface and is used as an effective alternative coagulation technique in various surgical disciplines. This trial aims to compare thermal injury in rat brain caused by APC and conventional bipolar coagulation technique. METHODS: A controlled study design with constant power setting and application time was established. Twenty rats were randomised into the APC and bipolar groups. Each group of ten rats had 20 treated lesions. Early and late histopathological changes, as well as maximum extent of the lesion after 48 hours (h) and 12 days were studied in overall 20 lesions. FINDINGS: Although the maximum depth of the lesions was different in APC (2.2 mm) and bipolar (1.8 mm) groups after 48 h, this did not achieve statistical significance (p=0.151). The superficially coagulated area was significantly larger after APC compared with the bipolar technique at the 48 h time point (p=0.032). After twelve days there were no differences in penetration depth (p=0.310) or coagulated area (p=0.222). CONCLUSION: Tissue defects after APC application on rat brains were comparable to conventional bipolar technique in this trial. The results suggest that argon plasma coagulation (APC) is an effective coagulation technique. PMID- 16708174 TI - The neuroprotective effect of inflammation: implications for the therapy of multiple sclerosis. AB - Autoreactive T cells are a regular component of the healthy immune system. It has been proposed that some of these autoreactive T cells even might have a protective function. Recent studies support this notion by demonstrating that: a) myelin-autoreactive T cells show neuroprotective effects in vivo, and b) activated antigen-specific human T cells and other immune cells produce bioactive brain-derived neurotrophic factor (BDNF) and other neurotrophic factors in vitro. Furthermore, BDNF is expressed in different types of inflammatory cells in brain lesions of patients with acute disseminated leukoencephalopathy or multiple sclerosis. It seems plausible that the immune cell-mediated import of BDNF and other neurotrophic factors into the central nervous system has functional consequences and implications for the therapy of multiple sclerosis and other neuroimmunological diseases. PMID- 16708175 TI - Intensive immunosuppression in multiple sclerosis. AB - Immunosuppressive drugs have been used out of label in multiple sclerosis (MS) for over 30 years and around 10% of patients are actually under immunosuppressive treatment. The rationale for immunosuppression in MS lies in the hypothesis that MS is an inflammatory immune-mediated disease that can take advantage of strong anti-inflammatory activity. Azathioprine, methotrexate, cyclophosphamide and mitoxantrone are the most utilised agents, but only the latter has been approved for clinically active MS. Many of them are safe in combination with interferon beta and are under investigation in controlled trials. Plasma exchange is limited to catastrophic attacks in refractory MS whilst bone marrow transplantation is considered in patients with an extremely severe, active disease as the final option in escalation therapy. Although immunosuppressants are best effective in induction therapy, their use is limited by toxicity and potential long-term risk. PMID- 16708176 TI - Neurobiological bases of rehabilitation. AB - The adult brain maintains the ability to reorganise throughout life. Motor cortical representations can reorganise rapidly in response to different stimuli. Important mechanisms for mediating reorganisation in the cerebral cortex involve the unmasking of existing, but latent, horizontal connections and modulation of GABAergic inhibition and synaptic efficacy. Interfering with these mechanisms can either block or enhance reorganisational processes. Following injury to the motor cortex alterations of the neurotransmitter system regulation, recruitment of additional undamaged brain areas even remote from the injury, and anatomical alterations such as axonal sprouting and synaptogenesis in the brain tissue surrounding the lesion or in the homotopic motor area of the non-affected hemisphere occur. The understanding of cortical reorganisation may enable us to apply principles of plasticity to the rehabilitation of patients after brain injury. PMID- 16708177 TI - Functional MRI to study brain plasticity in clinical neurology. AB - Functional magnetic resonance imaging (fMRI) is being widely used to study recovery of function in patients with several neurological conditions, including multiple sclerosis, stroke and Alzheimer's disease. The application of this MR technique has shown that plastic cortical changes do occur after central nervous system (CNS) injury of different aetiology, that such changes are related to the extent of CNS damage and that they can contribute in limiting the clinical consequences of brain damage. Conversely, the failure or exhaustion of the adaptive properties of the cerebral cortex might be among the factors responsible for the accumulation of 'fixed' neurological deficits. New studies aimed at investigating the effect of therapies devoted to promote brain plasticity are now warranted. PMID- 16708178 TI - Electrophysiological studies of brain plasticity of the motor system. AB - Brain plasticity, intended as the pattern of functional and structural changes in responses to environmental physiological or pathological events, is underlined by several mechanisms. Besides cortical changes such as changes in synaptic and representational patterns, morphologic and functional changes at the neuronal level have been hypothesised. Among the non-invasive techniques utilised in humans, electrophysiological methods allow description of the temporal dynamics of cortical activation and their inhibitory or excitatory functional significance. We will focus on the main applications of these techniques, mainly electroencephalography and transcranial magnetic stimulation, in the study of brain plasticity in humans following motor learning or lesions of the nervous system. PMID- 16708179 TI - Nosographical aspects and neuropsychological correlates of dementia. AB - The role of neuropsychological assessment in the clinical diagnosis and management of dementia has been shifting in recent years from an ancillary diagnostic test to an important tool for pre-clinical and differential diagnosis. Additionally, in conjunction with functional imaging and quantitative structural imaging, it is providing new insights into the cerebral correlates of cognition and behaviour. PMID- 16708180 TI - Genetics and neurobiology of frontotemporal lobar degeneration. AB - Frontotemporal dementia (FTD) is characterised by brain intracellular deposition of abnormally phosphorylated tau protein, considered responsible for neuronal death. Several familial cases with different mutations in the tau encoding gene (MAPT), located on chromosome 17, have been described. Besides, in a Danish family, the genetic defect has been associated to chromosome 3. Although many FTD families exhibit known mutations, in some cases none of them occur. Recent findings demonstrate an increased intrathecal production of both pro- and anti inflammatory cytokines in sporadic FTD patients. Besides, increased cerebrospinal fluid monocyte chemotactic protein-1 and interleukin-8 levels have been observed in FTD, whereas interferon-gamma-inducible protein-10 levels were similar to controls. PMID- 16708181 TI - Frontotemporal dementia. AB - Frontotemporal dementia is a clinicopathological syndrome caused by progressive degeneration of the frontal lobes, anterior temporal lobes or both. A wide spectrum of cognitive, psychological and behavioural symptoms has been reported, with early disturbance of personality and social conduct, impairment of executive functions and language, while memory and visuo-spatial skills are usually preserved. Published criteria can lead to a good accuracy in the antemortem diagnosis of frontotemporal dementia with a sensitivity of 85% and a specificity of 99%. Treatment rests on the management of behavioural disturbances. PMID- 16708182 TI - Vascular dementia. AB - Epidemiological, clinical, neuropsychological and neuroimaging features characterise vascular dementia (VaD) from other dementias of non-Alzheimer type. VaD does not represent a single entity, but quite a large group of conditions characterised by different pathophysiological correlates and various clinical findings. Subcortical VaD, in particular, is considered as a model of VaD because its well-defined diagnostic criteria allow identification of a homogeneous category for a better approach to investigation in diagnosis and treatment of vascular dementia. PMID- 16708183 TI - Parkinson's disease and dementia. AB - Parkinson's disease (PD) is one of the most common neurodegenerative disorders, affecting about 1% of the population over the age of 60. In addition to motor abnormalities, there are several non-motor signs and symptoms that may create a considerable burden for patients and care-givers. Dementia is common and affects approximately 40% of PD patients during the course of the disease, the risk for the development of dementia being 6 times higher than in non-PD age-matched controls. In most cases, PD patients with dementia (PDD) display a dysexecutive syndrome and visuospatial deficits, while memory is relatively unaffected. The overlap between PDD and dementia with Lewy bodies suggests that they likely share similar underlying neuropathological processes. PMID- 16708184 TI - Neuropathology of the dementias other than Alzheimer's. AB - Dementia is due to lesions destroying a large amount of circuits anatomically connected with, or functionally related to, the associative areas of the cortex and the limbic structures. Dementia is not only the clinical hallmark of neurodegenerative diseases, most often proteinopathies, primarily involving the cerebral cortex, but also a symptom frequently associated to movement disorders, or secondary to systemic or neurological, non-degenerative diseases. In the elderly, it is often the consequence of the cumulative effect of different diseases. PMID- 16708185 TI - Guillain-Barre syndrome. AB - Guillain-Barre syndrome (GBS) is an autoimmune acute peripheral neuropathy. Frequently a flu-like episode or a gastroenteritis precede GBS, and the cross reactivity between microbial and neural antigens partly explains the pathophysiology of the disease and the possible detection of antiganglioside antibodies. The weakness reaches its nadir in 2-4 weeks: the patients may be chair- or bed-bound, may need artificial ventilation and frequently experience dysautonomic dysfunction; 5-15% of the patients die and more patients are left with a disabling motor deficit and/or fatigue. Electrophysiology and cerebrospinal fluid evaluation support the diagnosis. The treatment of GBS is multidisciplinary, and both plasma exchange and high dose immunoglobulin (IVIg) are effective in reducing both the severity of the disease and the residual deficits. Finally, steroids are not effective in GBS. PMID- 16708186 TI - Status epilepticus. AB - Status epilepticus (SE) is a medical emergency. It requires prompt and adequate diagnosis and treatment, as it may induce CNS injury. It is mainly distinguished into generalised and partial SE on the basis of its major clinical features. There are very few data about SE physiopathology, but it is generally characterised by increasing unresponsiveness to treatment. SE diagnosis is based on EEG recording, associated with neuroimaging techniques and laboratory assays to detect underlying pathologies. During SE we distinguish three different conditions: initial, defined and refractory. Benzodiazepines represent first-line treatment, followed by phenytoin. Refractory SE requires ICU treatment to perform general anaesthesia. PMID- 16708187 TI - Acute encephalopathies. AB - In daily hospital activity a frequent observation is that of patients in conditions of acute global suffering of the central nervous system due to a primary cause or symptomatic of other diseases. This condition shows an alteration of the state of consciousness, which involves three possible semeiologic manifestations: delirium (acute confusional state), acute confusional state associated with psychomotor underactivity and coma. The possible causes of an acute encephalopathy (AE) are: metabolic, iatrogenic and toxic, infective, primary neurologic, surgery-related and others. Diagnostic-therapeutic routes to be followed by emergency room doctors and neurologists dealing with AE are defined. PMID- 16708188 TI - The stroke in an emergency: the grey areas. AB - In spite of the development of advanced technologies, diagnosis and therapy for an ictus in emergency still pose difficulties in a significant percentage of cases. Diagnostic problems are encountered both when faced with focal symptoms and in the presence of disturbances to consciousness, confusional states, vertigo or cephalea. The most significant therapeutic problems, on the other hand, are encountered in the case of extended cardioembolic ischaemias, haemorrhages from venous thrombosis and intradural arterial dissections (eventual antithrombotic treatment), when faced with a large cerebral oedema (eventual medical and/or surgical treatment), or in indicating surgery for intraparenchymal haemorrhages or symptomatic stenotic carotids. PMID- 16708189 TI - The management of headaches in the emergency department: critical issues. AB - Typical cases of the most common kinds of headache can be diagnosed and treated by general practitioners (GPs). Non-traumatic patients with de novo acute sudden onset disabling headaches as well as significant worsening of pre-existing headaches seek care at emergency departments (EDs) and represent a diagnostic challenge for the consultant neurologist, who is the specialist of reference for the entire diagnostic process. Explicit diagnostic criteria for the classification of headache disorders (ICHD-II) are fundamental for verifying the final diagnosis, but in the emergency setting diagnostic and therapeutic guidelines and recommendations, coupled with lists of diagnostic alarms and warnings, may further contribute to the preliminary identification of secondary headaches. PMID- 16708191 TI - Early treatment. AB - Class I clinical trials demonstrated that immunomodulatory treatments (interferon beta and glatiramer acetate) reduce the disease activity and the accumulation of disability in relapsing-remitting multiple sclerosis (MS). Moreover, interferon beta-1b had similar positive effects also in secondary progressive MS. The magnitude of these clinical effects was modest, but the reduction of inflammatory activity, as revealed by magnetic resonance imaging, was marked. There is converging evidence from new pathological studies and from new magnetic resonance techniques, characterised by an increased pathological specificity, that already in the early phases of the disease the inflammatory activity determines irreversible axonal damage. Moreover, the amount of inflammatory activity at clinical presentation of the disease has some value for predicting long-term disability. Taken together, these data indicate that patients may benefit from early treatment; the positive results of three double-blind placebo-controlled clinical trials (Early Treatment of Multiple Sclerosis and Controlled High Risk Subjects Avonex Multiple Sclerosis Prevention Study and BENEFIT) support this conclusion. PMID- 16708192 TI - Conversion of agricultural feedstock and coproducts into poly(hydroxyalkanoates). AB - Aside from their importance to the survival and general welfare of mankind, agriculture and its related industries produce large quantities of feedstocks and coproducts that can be used as inexpensive substrates for fermentative processes. Successful adoption of these materials into commercial processes could further the realization of a biorefinery industry based on agriculturally derived feedstocks. One potential concept is the production of poly(hydroxyalkanoate) (PHA) polymers, a family of microbial biopolyesters with a myriad of possible monomeric compositions and performance properties. The economics for the fermentative production of PHA could benefit from the use of low-cost agricultural feedstocks and coproducts. This mini-review provides a brief survey of research performed in this area, with specific emphasis on studies describing the utilization of intact triacylglycerols (vegetable oils and animal fats), dairy whey, molasses, and meat-and-bone meal as substrates in the microbial synthesis of PHA polymers. PMID- 16708193 TI - Square-plate culture method allows detection of differential gene expression and screening of novel, region-specific genes in Aspergillus oryzae. AB - When grown on solid agar medium, the mycelium of a filamentous fungus, Aspergillus oryzae, forms three morphologically distinct regions: the tip (T), white (W), and basal (B) regions. In this study, we developed the square-plate culture method, a novel culture method that enabled the extraction of mRNA samples from the three regions and analyzed the differential gene expression of the A. oryzae mycelium in concert with the microarray technique. Expression of genes involved in protein synthesis was predominant in the T region; relative expression was, at most, six times higher in the T region compared to the other regions. Genes encoding hypothetical proteins were expressed at high levels in the W and B regions. In addition, genes coding transporters/permeases were predominantly transcribed in the B region. By analyzing the expression patterns of genes in the three regions, we demonstrated the dynamic changes in the regulation of gene expression that occur along the mycelium of filamentous fungi. Consequently, our study established a method to analyze and screen for region specific genes whose function may be essential for morphogenesis and differentiation in filamentous fungi and whose traits may be beneficial to the biotechnology industry. PMID- 16708194 TI - Isolation and characterization of a novel Burkholderia cepacia with strong antifungal activity against Rhizoctonia solani. AB - Strain CF-66 with strong antifungal activity against Rhizoctonia solani was isolated from compost samples. It is clearly demonstrated that strain CF-66 is belonging to Burkholderia cepacia complex by the morphological and biochemical tests and 16S rDNA sequence. The B. cepacia complex consists of a group of bacteria currently organized into nine genomovars, among them genomovar II and genomovar III, contain the highly epidemic strains. However, it was known that strain CF-66 is not a member of genomovar II or III of the B. cepacia complex by species-specific polymerase chain reaction assay. In this study, the antifungal compound CF66I produced by strain CF-66 was purified and characterized. Based on the nuclear magnetic resonance, GC-MS spectral and infrared spectral data, CF66I was confirmed to have amide bonds, alpha-methyl fatty acid, bromine, and some structural units such as CH(2)CH(2)O. CF66I is stable to high temperature, proteolytic enzymes, and organic solvents. CF66I inhibit the growth of a variety of plant pathogenic fungi and pathogenic yeast, whereas bacterial cells are unaffected. CF66I mainly reduced hyphal extension rates in a dose-dependent manner and induced severe change in cell morphology that resulted in swelled and formed very short hyphae with multiple branches. PMID- 16708195 TI - Construction of ivermectin producer by domain swaps of avermectin polyketide synthase in Streptomyces avermitilis. AB - Ivermectin, 22, 23-dihydroavermectin B1, is commercially important in human, veterinary medicine, and pesticides. It is currently synthesized by chemical reduction of the double bond between C22 and C23 of avermectins B1, which are a mixture of B1a (>80%) and B1b (<20%) produced by fermentation of Streptomyces avermitilis. The cost of ivermectin is much higher than that of avermectins B1 owing to the necessity of region-specific hydrogenation at C22-C23 of avermectins B1 with rhodium chloride as the catalyst for producing ivermectin. Here we report that ivermectin can be produced directly by fermentation of recombinant strains constructed through targeted genetic engineering of the avermectin polyketide synthase (PKS) in S. avermitilis Olm73-12, which produces only avermectins B and not avermectins A and oligomycin. The DNA region encoding the dehydratase (DH) and ketoreductase (KR) domains of module 2 from the avermectin PKS in S. avermitilis Olm73-12 was replaced by the DNA fragment encoding the DH, enoylreductase, and KR domains from module 4 of the pikromycin PKS of Streptomyces venezuelae ATCC 15439 using a gene replacement vector pXL211. Twenty seven of mutants were found to produce a small amount of 22, 23-dihydroavermectin B1a and avermectin B1a and B2a by high performance liquid chromatography and liquid chromatography mass spectrometry analysis. This study might provide a route to the low-cost production of ivermectin by fermentation. PMID- 16708197 TI - Determination of antimicrobial residues and metabolites in the aquatic environment by liquid chromatography tandem mass spectrometry. AB - Antimicrobials are used in large quantities in human and veterinary medicine. Their environmental occurrence is of particular concern due to the potential spread and maintenance of bacterial resistance. After intake by the organisms, the unchanged drug and its metabolized forms are excreted and enter wastewater treatment plants where they are mostly incompletely eliminated, and are therefore eventually released into the aquatic environment. The reliable detection of several antimicrobials in different environmental aqueous compartments is the result of great improvements achieved in analytical chemistry. This article provides an overview of the more outstanding analytical methods based on liquid chromatography tandem mass spectrometry, developed and applied to determine antimicrobial residues and metabolites present in surface, waste, and ground waters. PMID- 16708200 TI - [Unclear peranal blood loss in a 21-year-old man]. PMID- 16708201 TI - [Lumbar disc arthroplasty: indications, biomechanics, types, and radiological criteria]. AB - Lumbar total disc replacement (TDR) was developed to treat a painful degenerative lumbar motion segment while avoiding the disadvantages of fusion surgery, such as adjacent segment instabilities. Early clinical results with TDR have shown a significant reduction in low back pain and a significant improvement in disability scores. When compared to fusion, the results with TDR tend to be superior in the short-term follow-up and initial rehabilitation is faster. The radiological assessment is an integral part of the preoperative work-up. Plain X rays of the lumbar spine should be complemented by flexion - extension views in order to assess residual segmental mobility. Computed tomography is used to exclude osteoarthritis of the zygapophyseal joints, Baastrup's disease (kissing spines) and other sources of low back pain. Magnetic resonance imaging is useful to exclude substantial disc protrusions; it allows for the detection of disc dehydration and bone marrow edema in the case of activated spondylochondrosis. If osteoporosis is suspected, an osteodensitometry of the lumbar spine should be performed. Postoperative plain X-rays should include antero-posterior and lateral views as well as flexion - extension views in the later postoperative course. Measurements should determine the disc space height in the lateral view, the segmental and total lumbar lordosis as well as the segmental mobility in the flexion - extension views. The ideal position of a TDR is exactly central in the ap-view and close to the dorsal border of the vertebral endplates in the lateral view. Malpositioning may cause segmental hyperlordosis and unbalanced loading of the endplates with the risk of implant subsidence and migration. PMID- 16708205 TI - Diffusion tensor imaging of midline posterior fossa malformations. AB - BACKGROUND: Diffusion tensor imaging and tractography have been used to evaluate a variety of brain malformations. However, these studies have focused mainly on malformations involving the supratentorial compartments. There is a paucity of data on diffusion tensor imaging of posterior fossa malformations. OBJECTIVE: To describe the color vector maps and modified or abnormal tracts of midline posterior fossa malformations. MATERIALS AND METHODS: Diffusion tensor imaging was performed in one patient with rhombencephalosynapsis and two with Joubert syndrome. Color vector maps of fractional anisotropy were used to place a region of interest for seed point of fiber tracking. RESULTS: The vermis was severely hypoplastic or absent in rhombencephalosynapsis and Joubert syndrome. In rhombencephalosynapsis, vertically oriented fibers were visualized in the midportion of the cerebellum. The location of the deep cerebellar nuclei could be inferred from the amiculum and were medially located in rhombencephalosynapsis. In the two patients with Joubert syndrome, the horizontally arranged superior cerebellar peduncles were well demonstrated on the color vector maps. Failure of the superior cerebellar peduncles to decussate in the mesencephalon was also well demonstrated on both color vector maps and tractography. The deep cerebellar nuclei were more laterally located in Joubert syndrome. CONCLUSION: The use of tractography in midline posterior fossa malformations expands our understanding of these malformations. PMID- 16708202 TI - Modified technique to create morphologically reproducible elastase-induced aneurysms in rabbits. AB - INTRODUCTION: The purpose of this study was to create morphologically reproducible elastase-induced model aneurysms in rabbits. METHODS: We created 120 elastase-induced aneurysms in rabbits using two different methods: the standard technique (group 1, n=62) and a modified technique (group 2, n=58). In the standard technique a small cutdown with a focal area of exposure of the mid-right common carotid artery (RCCA) was employed, while in the modified technique the RCCA was completely exposed to its origin. We measured aneurysm sizes (neck diameter, width and height) in the two groups. The aneurysm sizes were compared between the two groups using Student's t test, and the standard deviations of the aneurysm sizes were compared between the groups using the F test. RESULTS: The mean aneurysm neck size, width and height in group 1 were 3.4+/-1.2 mm, 3.8+/-1.0 mm and 8.0+/-1.7 mm, respectively, and in group 2, were 3.2+/-0.9 mm, 3.7+/-0.6 mm and 9.1+/-1.8 mm, respectively. The differences in mean aneurysm neck and width between the two groups were not significant (P>0.05). However, there were significant differences in the standard deviation of these two parameters between the two groups (P<0.05 and P<0.01, respectively). The mean aneurysm height in group 2 was larger than in group 1 (P<0.001), but no significant difference in the standard deviation of this parameter between the two groups was found (P>0.05). CONCLUSION: The results indicate that more consistent aneurysm diameters can be created using the modified technique. PMID- 16708206 TI - Calcified pulmonary thromboembolism in a child with sickle cell disease: value of multidetector CT in patients with acute chest syndrome. AB - The incidence of pulmonary embolism in children is not clearly known, but is believed to be low. Risk factors for pulmonary thromboembolism include central venous catheter, malignancy, surgery, infection, trauma, and congenital hypercoagulable disorders. Children with sickle cell disease are prothrombotic and are at an increased risk of thromboembolism. The incidence of this event is unknown because these children are often not thoroughly imaged. We report here a case of a calcified pulmonary thromboembolism in a child with sickle cell disease and emphasize the use of multidetector CT in detection of pulmonary thromboembolism in children with sickle cell disease. PMID- 16708207 TI - A periureteric venous ring diagnosed by MRI: an unusual cause of flank pain. AB - We describe a pediatric patient with upper tract collecting system dilatation caused by a periureteric venous ring. The diagnosis was made after MR urography. This rare inferior vena caval anomaly has not been documented in the pediatric literature, nor have the MRI appearances been described. MRI is ideally suited for the assessment of suspected retrocaval ureters in order to identify this variant so that an appropriate surgical approach can be planned. PMID- 16708210 TI - Direct analysis of a GPCR-agonist interaction by surface plasmon resonance. AB - Despite their clinical importance, detailed analysis of ligand binding at G protein coupled receptors (GPCRs) has proved difficult. Here we successfully measure the binding of a GPCR, neurotensin receptor-1 (NTS-1), to its ligand, neurotensin (NT), using surface plasmon resonance (SPR). Specific responses were observed between NT and purified, detergent-solublised, recombinant NTS-1, using a novel configuration where the biotinylated NT ligand was immobilised on the biosensor surface. This SPR approach shows promise as a generic approach for the study of ligand interactions with other suitable GPCRs. PMID- 16708211 TI - Voltage changes involving photosystem II quinone-iron complex turnover. AB - An electrometrical technique was used to investigate proton-coupled electron transfer between the primary plastoquinone acceptor Q (A) (-) and the oxidized non-heme iron Fe(3+) on the acceptor side of photosystem II core particles incorporated into phospholipid vesicles. The sign of the transmembrane electric potential difference Deltapsi (negative charging of the proteoliposome interior) indicates that the iron-quinone complex faces the interior surface of the proteoliposome membrane. Preoxidation of the non-heme iron was achieved by addition of potassium ferricyanide entrapped into proteoliposomes. Besides the fast unresolvable kinetic phase (tau approximately 0.1 micro s) of Deltapsi generation related to electron transfer between the redox-active tyrosine Y(Z) and Q(A), an additional phase in the submillisecond time domain (tau approximately 0.1 ms at 23 degrees C, pH 7.0) and relative amplitude approximately 20% of the amplitude of the fast phase was observed under exposure to the first flash. This phase was absent under the second laser flash, as well as upon the first flash in the presence of DCMU, an inhibitor of electron transfer between Q(A) and the secondary quinone Q(B). The rate of the additional electrogenic phase is decreased by about one-half in the presence of D(2)O and is reduced with the temperature decrease. On the basis of the above observations we suggest that the submillisecond electrogenic reaction induced by the first flash is due to the vectorial transfer of a proton from external aqueous phase to an amino acid residue(s) in the vicinity of the non-heme iron. The possible role of the non-heme iron in cyclic electron transfer in photosystem II complex is discussed. PMID- 16708212 TI - Localized pigmented villonodular synovitis of the knee: acute onset in pregnancy. AB - A pregnant patient in the first trimester presented with acute onset knee pain and effusion. As the clinical status was mimicking septic arthritis, surgery was performed. Arthroscopy demonstrated a local synovial tumor and excision was performed with arthrotomy. Microscopic evaluation revealed localized pigmented villonodular synovitis. Due to the presence of necrosis and hematoma in the tumor, we hypothesize that, that torsion or bleeding of the tumor in the presence of physiological pregnancy- related metabolic changes might have been the cause of acute presentation. PMID- 16708213 TI - Prognostic factors in patients with recently diagnosed incurable cancer: a systematic review. AB - GOALS OF WORK: To review the literature and develop a conceptual framework about prognostic factors for people presenting to medical oncologists with recently diagnosed incurable cancer. MATERIALS AND METHODS: Medline was searched from January 2000 to October 2003 to identify articles testing associations between clinical or laboratory variables and survival time in adults with advanced solid tumours and median survival of 3 to 24 months. We recorded how frequently prognostic factors were significantly associated with survival in univariable and multivariable analyses. RESULTS: There were 53 studies included. The factors associated with survival were organised into four categories related to attributes of the host the tumour, the treatment and the interactions between host, tumour and treatment (symptoms, quality of life, performance status and laboratory tests). Co-morbidity was consistently associated with shorter survival. Age and gender were not consistently associated with survival duration, except in lung cancer where females survived longer. Tumour-related factors associated with shorter survival included primary tumour (lung), metastatic site (liver, brain and visceral) and disease extent. Symptoms associated with shorter survival included those of the anorexia-cachexia syndrome, dyspnoea, pain and impaired physical well being. Performance status was strongly associated with survival in most studies. Laboratory tests associated with shorter survival included anaemia, thrombocytopenia, hypoalbuminaemia and elevated serum levels of both alkaline phosphatase and lactate dehydrogenase. CONCLUSION: Prognostic factors in patients with advanced cancer can be conceptualised as attributes of the host, tumour, treatment and interactions between the three reflected in symptoms, quality of life performance status and laboratory tests. PMID- 16708214 TI - Effects of anthracene on development of an arbuscular mycorrhizal fungus and contribution of the symbiotic association to pollutant dissipation. AB - The influence of anthracene, a low molecular weight polycyclic aromatic hydrocarbon (PAH), on chicory root colonization by Glomus intraradices and the effect of the root colonization on PAH degradation were investigated in vitro. The fungus presented a reduced development of extraradical mycelium and a decrease in sporulation, root colonization, and spore germination when exposed to anthracene. Mycorrhization improved the growth of the roots in the medium supplemented containing 140 mg l(-1) anthracene, suggesting a positive contribution of G. intraradices to the PAH tolerance of roots. Anthracene disappearance from the culture medium was quantified; results suggested that nonmycorrhizal chicory roots growing in vitro were able to contribute to anthracene dissipation, and in addition, that mycorrhization significantly enhanced anthracene dissipation. These monoxenic experiments demonstrated a positive contribution of the symbiotic association to anthracene dissipation in the absence of other microorganisms. In addition to anthracene dissipation, intracellular accumulation of anthracene was detected in lipid bodies of plant cells and fungal hyphae, indicating intracellular storage capacity of the pollutant by the roots and the mycorrhizal fungus. PMID- 16708216 TI - Interobserver agreement on the radiological criteria of the International Panel on the diagnosis of multiple sclerosis. AB - We assessed the interobserver agreement on the radiological part of the International Panel (IP) criteria for the diagnosis of multiple sclerosis (MS), comprising the assessment of dissemination in space (DIS) and time (DIT) based exclusively on MRI. Four radiologists trained and four radiologists naive in the application of the IP criteria scored the fulfillment for DIS (i.e., > or =3 periventricular, > or =1 infratentorial, > or =1 juxtacortical, > or = 1 enhancing lesion or a total of > or =9 T2-weighted lesions) and DIT (presence of new or enhancing lesions at follow-up) in baseline and two follow-up scans from 20 patients suspected for having MS. The IP-trained radiologists agreed at least moderately on all assessments (kappa>0.40), whereas the IP-naive radiologists showed fair agreement (kappa<0.40) on five of 16 assessments. In the final conclusion on DIS and DIT, the IP-trained radiologists agreed substantially on both items (kappa=0.62 and kappa=0.60, respectively) compared with a fair agreement on DIS (kappa=0.29) and moderate agreement on DIT (kappa=0.52) among the IP-naive radiologists. Given the poor interobserver agreement among IP-naive observers, the new IP criteria for MS require additional training and should perhaps be simplified. PMID- 16708218 TI - Hepatocellular adenoma: findings at state-of-the-art magnetic resonance imaging, ultrasound, computed tomography and pathologic analysis. AB - The purpose of this paper is to describe the most recent concepts and pertinent findings of hepatocellular adenomas, including clinical presentation, gross pathology and histology, pathogenesis and transformation into hepatocellular carcinoma (HCC), and imaging findings at ultrasound (US), computed tomography (CT), and magnetic resonance (MR) imaging. PMID- 16708217 TI - Testicular calcification and microlithiasis: association with primary intra testicular malignancy in 3,477 patients. AB - The prevalence of all forms of scrotal and testicular calcification and their association with testicular tumour in a symptomatic paediatric and adult population was investigated. A retrospective study of all testicular ultrasound examinations performed at a single centre over a 5-year period was undertaken. All studies were performed by experienced operators, recorded in a standard method, using high-frequency linear array transducers (> or =10 MHz). All available images (95.2%) were reviewed by experienced operators, recording the location and type of scrotal and testicular calcification according to a pre determined schedule. A total of 3,854 studies were reviewed on 3,477 patients (age range: 1 month to 91 years). In the adult group, 3,279 examinations were analysed. Prevalence of testicular microlithiasis (TM) was 2.0%, and the prevalence of other non-microlithiasis testicular calcification (non-TM calcification) was 1.7%. Testicular tumour was associated with TM (odds ratio 9.5, P<0.001) and non-TM calcification (odds ratio 11.4, P<0.001) but not with other types of scrotal calcification. A total of 198 paediatric examinations were analysed. Prevalence of TM was 2.0% and the prevalence of non-TM calcification was 0.5%. One tumour (lymphoma) was identified, with no associated calcification. This study confirms the reported association between TM and testicular tumour and finds a previously unreported association between non-TM calcification and testicular tumour. PMID- 16708219 TI - Intracranial 2D and 3D DSA with flat panel detector of the direct conversion type: initial experience. AB - The purpose of this study was to compare the image quality of two-dimensional (2D) digital subtraction angiography (DSA) between a flat panel detector (FPD) of the direct conversion type with low radiation dose and a conventional image intensifier (I.I.)-TV system, and to assess 3D DSA with the FPD system in the depiction of intracranial vessels. Fifteen consecutive patients (five men, ten women; age range: 18-82 years; mean age: 55.5 years) were prospectively included in this study. All patients underwent 2D DSA with both the FPD and I.I.-TV system in one projection. The radiation doses during angiography were evaluated using a phantom. The 3D DSA images were created from the rotational DSA data with the FPD system. Two blinded radiologists independently evaluated 2D DSA with the FPD system and I.I.-TV system using a 5-point assessment scale (excellent to not visible) to assess the depiction of intracranial vessels. MIP and volume rendering (VR) images of 3D DSA with the FPD system were also evaluated using a 5 point scale (excellent to not visible). DSA and fluoroscopy dose measurements with the phantom showed a dose reduction of approximately 85% and 9% with the FPD system compared with the I.I.-TV system, respectively. For 2D DSA, the FPD system was significantly superior to the I.I.-TV system with respect to the visibility of the peripheral and perforating vessels (p<0.05). The peripheral and perforating vessels were also sufficiently visualized on MIP images of 3D DSA in all 15 cases. Our FPD system was found to be superior to the I.I.-TV system in visualizing small intracranial vessels combined with a significant reduction of radiation dose, and was able to create high-quality 3D DSA images on which high spatial resolution allowed precise visualization of small vessels such as perforating ones. PMID- 16708220 TI - Radial scars diagnosed at stereotactic core-needle biopsy: surgical biopsy findings. AB - We retrospectively reviewed surgical biopsy findings of lesions diagnosed as radial scars (RS) at stereotactic core-needle biopsy (SCNB). RS was diagnosed in 52 of 1415 (3.7%) consecutive mammographically detected lesions that underwent 14 gauge automated SCNB. Subsequent surgical biopsy findings were available for 43 lesions in 41 women constituting the study group. Of these 43 lesions, histopathological analysis of the surgical specimen yielded RS in 27 (63%), RS plus atypical ductal hyperplasia (ADH) in 8 (18%), RS plus carcinoma in five (12%), and only carcinoma in three (7%). Carcinomas underestimated at SCNB (n=8) were two in situ carcinomas, two invasive ductal carcinomas not otherwise specified, and four tubular carcinomas. A statistically significant difference (P=0.02) was found between the mean pathologic size of RS without carcinoma and of RS containing carcinoma. Mammographic features could not be used reliably to predict the presence of carcinoma at excision of lesions diagnosed as RS at SCNB. The results suggest that the diagnosis of RS at 14-gauge SCNB of mammographically detected lesions is an indication for surgical biopsy because of the high prevalence of carcinoma in these lesions. PMID- 16708221 TI - Management of a congenital H-type anorectal fistula by anterior sagittal anorectovaginoplasty (ASARVP). PMID- 16708222 TI - Colony-stimulating factor-1 receptor gene polymorphisms and Crohn's disease. PMID- 16708223 TI - Liver capsule rupture in eclampsia: treatment with hepatic artery embolisation. AB - INTRODUCTION: Spontaneous hepatic rupture associated with pregnancy is a rare and very serious complication associated with eclampsia or pre-eclampsia. CASE REPORT: We report a case of hepatic rupture following eclampsia at 27 weeks gestation which was managed successfully by selective hepatic artery embolisation as an adjunct to liver packing. CONCLUSION: Selective embolisation of the hepatic artery is an effective means of therapy for hepatic artery bleeding and is an adjunct to liver packing in spontaneous hepatic rupture in eclamptic patients. PMID- 16708224 TI - Cavernous hemangiomatous polyp: an unusual case of perimenopausal bleeding. AB - INTRODUCTION: Significant vascular malformations of uterus are rare but can cause serious menorrhagia. CASE REPORT: We present the case of a 40-year-old multiparous woman who presented with perimenopausal bleeding due to cavernous hemangiomatous polyp successfully treated by polypectomy. CONCLUSION: Cavernous hemangioma can be a rare cause of perimenopausal bleeding. The same found in a polyp may be treated successfully by polypectomy. PMID- 16708225 TI - Microgravity effects on leaf morphology, cell structure, carbon metabolism and mRNA expression of dwarf wheat. AB - The use of higher plants as the basis for a biological life support system that regenerates the atmosphere, purifies water, and produces food has been proposed for long duration space missions. The objective of these experiments was to determine what effects microgravity (microg) had on chloroplast development, carbohydrate metabolism and gene expression in developing leaves of Triticum aestivum L. cv. USU Apogee. Gravity naive wheat plants were sampled from a series of seven 21-day experiments conducted during Increment IV of the International Space Station. These samples were fixed in either 3% glutaraldehyde or RNAlater or frozen at -25 degrees C for subsequent analysis. In addition, leaf samples were collected from 24- and 14-day-old plants during the mission that were returned to Earth for analysis. Plants grown under identical light, temperature, relative humidity, photoperiod, CO(2), and planting density were used as ground controls. At the morphological level, there was little difference in the development of cells of wheat under microg conditions. Leaves developed in mug have thinner cross-sectional area than the 1g grown plants. Ultrastructurally, the chloroplasts of microg grown plants were more ovoid than those developed at 1g, and the thylakoid membranes had a trend to greater packing density. No differences were observed in the starch, soluble sugar, or lignin content of the leaves grown in microg or 1g conditions. Furthermore, no differences in gene expression were detected leaf samples collected at microg from 24-day-old leaves, suggesting that the spaceflight environment had minimal impact on wheat metabolism. PMID- 16708226 TI - A complex rearrangement on chromosome 22 affecting both homologues; haplo insufficiency of the Cat eye syndrome region may have no clinical relevance. AB - The presence of highly homologous sequences, known as low copy repeats, predisposes for unequal recombination within the 22q11 region. This can lead to genomic imbalances associated with several known genetic disorders. We report here a developmentally delayed patient carrying different rearrangements on both chromosome 22 homologues, including a previously unreported rearrangement within the 22q11 region. One homologue carries a deletion of the proximal part of chromosome band 22q11. To our knowledge, a 'pure' deletion of this region has not been described previously. Four copies of this 22q11 region, however, are associated with Cat eye syndrome (CES). While the phenotypic impact of this deletion is unclear, familial investigation revealed five normal relatives carrying this deletion, suggesting that haplo-insufficiency of the CES region has little clinical relevance. The other chromosome 22 homologue carries a duplication of the Velocardiofacial/DiGeorge syndrome (VCFS/DGS) region. In addition, a previously undescribed deletion of 22q12.1, located in a relatively gene-poor region, was identified. As the clinical features of patients suffering from a duplication of the VCFS/DGS region have proven to be extremely variable, it is impossible to postulate as to the contribution of the 22q12.1 deletion to the phenotype of the patient. Additional patients with a deletion within this region are needed to establish the consequences of this copy number alteration. This study highlights the value of using different genomic approaches to unravel chromosomal alterations in order to study their phenotypic impact. PMID- 16708227 TI - Spatial refuge from intraguild predation: implications for prey suppression and trophic cascades. AB - The ability of predators to elicit a trophic cascade with positive impacts on primary productivity may depend on the complexity of the habitat where the players interact. In structurally-simple habitats, trophic interactions among predators, such as intraguild predation, can diminish the cascading effects of a predator community on herbivore suppression and plant biomass. However, complex habitats may provide a spatial refuge for predators from intraguild predation, enhance the collective ability of multiple predator species to limit herbivore populations, and thus increase the overall strength of a trophic cascade on plant productivity. Using the community of terrestrial arthropods inhabiting Atlantic coastal salt marshes, this study examined the impact of predation by an assemblage of predators containing Pardosa wolf spiders, Grammonota web-building spiders, and Tytthus mirid bugs on herbivore populations (Prokelisia planthoppers) and on the biomass of Spartina cordgrass in simple (thatch-free) and complex (thatch-rich) vegetation. We found that complex-structured habitats enhanced planthopper suppression by the predator assemblage because habitats with thatch provided a refuge for predators from intraguild predation including cannibalism. The ultimate result of reduced antagonistic interactions among predator species and increased prey suppression was enhanced conductance of predator effects through the food web to positively impact primary producers. Behavioral observations in the laboratory confirmed that intraguild predation occurred in the simple, thatch-free habitat, and that the encounter and capture rates of intraguild prey by intraguild predators was diminished in the presence of thatch. On the other hand, there was no effect of thatch on the encounter and capture rates of herbivores by predators. The differential impact of thatch on the susceptibility of intraguild and herbivorous prey resulted in enhanced top down effects in the thatch-rich habitat. Therefore, changes in habitat complexity can enhance trophic cascades by predator communities and positively impact productivity by moderating negative interactions among predators. PMID- 16708228 TI - Nitrogen response efficiency increased monotonically with decreasing soil resource availability: a case study from a semiarid grassland in northern China. AB - The concept of nutrient use efficiency is central to understanding ecosystem functioning because it is the step in which plants can influence the return of nutrients to the soil pool and the quality of the litter. Theory suggests that nutrient efficiency increases unimodally with declining soil resources, but this has not been tested empirically for N and water in grassland ecosystems, where plant growth in these ecosystems is generally thought to be limited by soil N and moisture. In this paper, we tested the N uptake and the N use efficiency (NUE) of two Stipa species (S. grandis and S. krylovii) from 20 sites in the Inner Mongolia grassland by measuring the N content of net primary productivity (NPP). NUE is defined as the total net primary production per unit N absorbed. We further distinguished NUE from N response efficiency (NRE; production per unit N available). We found that NPP increased with soil N and water availability. Efficiency of whole-plant N use, uptake, and response increased monotonically with decreasing soil N and water, being higher on infertile (dry) habitats than on fertile (wet) habitats. We further considered NUE as the product of the N productivity (NP the rate of biomass increase per unit N in the plant) and the mean residence time (MRT; the ratio between the average N pool and the annual N uptake or loss). The NP and NUE of S. grandis growing usually in dry and N-poor habitats exceeded those of S. krylovii abundant in wet and N-rich habitats. NUE differed among sites, and was often affected by the evolutionary trade-off between NP and MRT, where plants and communities had adapted in a way to maximize either NP or MRT, but not both concurrently. Soil N availability and moisture influenced the community-level N uptake efficiency and ultimately the NRE, though the response to N was dependent on the plant community examined. These results show that soil N and water had exerted a great impact on the N efficiency in Stipa species. The intraspecific differences in N efficiency within both Stipa species along soil resource availability gradient may explain the differences in plant productivity on various soils, which will be conducive to our general understanding of the N cycling and vegetation dynamics in northern Chinese grasslands. PMID- 16708232 TI - Patient satisfaction following carpal-tunnel decompression: a comparison of patients with and without osteoarthritis of the wrist. AB - The aim of this study was to assess whether surgical decompression for carpal tunnel syndrome (CTS) in the presence of primary or secondary osteoarthritis of the wrist is associated with poorer patient satisfaction. We did a retrospective matched cohort study. Twenty-four patients who underwent surgical decompression for CTS secondary to osteoarthritis were identified by reviewing the notes and the radiographs. A control group consisted of 24 patients without osteoarthritis of the wrist who underwent carpal-tunnel decompression. The control group was matched for age, sex, side, and neuro-physiological severity of the nerve compression. In the group with osteoarthritis of the wrist, 17 (71%) patients reported their symptom relief as satisfactory, and 7 (29%) reported the results as unsatisfactory. In the control group, 23 (96%) patients reported their symptom relief as satisfactory, and 1 (4%) reported the results as unsatisfactory (P=0.0325). In conclusion, patient satisfaction following surgical decompression in patients with secondary CTS due to osteoarthritis was significantly lower compared to patients without osteoarthritis of the wrist. PMID- 16708233 TI - Long-term results of ventro-dorsal versus ventral instrumentation fusion in the treatment of spondylitis. AB - This study examines prospectively the randomised, long-term, clinical and radiological results of the treatment of spondylitis patients by ventro-dorsal or ventral spine fusion. Group 1 consisted of 12 patients who (after ventral removal of the focus of infection and autologous bone grafting) were treated by dorsal instrumentation. Group 2 consisted of ten patients who, after similar ventral removal and bone interposition, were stabilised by ventral instrumentation. The patients prospectively underwent clinical and radiological studies. In addition, they were asked to fill in self-assessment questionnaires such as the short-form (SF)-36 health survey, the Oswestry questionnaire, and the visual analog scales (VAS). The postoperative follow-ups were at 6 months, 2 years and 5.4 years. It proved possible to demonstrate clinically that patients with an isolated ventral spondylodesis feel significantly better and experience significantly less pain in the area of spinal fusion than patients with ventro-dorsal fusion 2 and 5.4 years after the operation. Over a number of years a stable fusion can be achieved through either operation. Ventral stabilisation yields more advantages than dorsal instrumentation in the long term. These advantages result in a clinically smoother course after the operation. If, in the individual case, ventral instrumentation is feasible, this method should be used. PMID- 16708234 TI - Amifostine and glutathione prevent ifosfamide- and acrolein-induced hemorrhagic cystitis. AB - INTRODUCTION: Ifosfamide (IFS) is an antineoplastic alkylating agent whose major side effect is hemorrhagic cystitis (HC). This toxicity is attributed to the renal excretion of acrolein (ACR), a highly urotoxic IFS metabolite. Despite the clinical use of mesna to prevent HC, a significant percent ( approximately 33%) of patients present with at last one feature of HC, mainly hematuria. AIM: To investigate the use of two antioxidants-amifostine and glutathione-for the prevention of experimental IFS- and ACR-induced HC. MATERIALS AND METHODS: Male Swiss mice were treated intraperitoneal (i.p.) with saline (control), glutathione (125, 250 or 500 mg/kg) or amifostine (25, 50 or 100 mg/kg), and 30 min later they received a single i.p. injection of IFS at a dose of 400 mg/kg. To investigate the systemic effects of the antioxidants on ACR-induced HC, the animals were treated with saline, amifostine (50 mg/kg, i.p.) or glutathione (500 mg/kg, i.p.), and 30 min afterward with 75 mug ACR intravesically (i.ve.). In another set of experiments, the antioxidants were injected directly into the bladder, where the mice received a single i.ve injection of ACR (75 mug) plus amifostine (1.5 mg/kg) or glutathione (2 mg/kg). HC was measured 3 h after IFS or ACR injection according to bladder wet weight, macroscopic (edema and hemorrhage) and microscopic changes, i.e., edema, hemorrhage, cellular infiltration, fibrin deposition and urothelial desquamation. RESULTS: Pretreatments with amifostine or glutathione prevented IFS-induced HC in a dose-dependent manner. Furthermore, ACR induced HC was also prevented by systemic (i.p.) or local (i.ve.) pretreatment with glutathione or amifostine. The greatest protective effect was seen with local amifostine treatment (2 mg/kg i.ve.) (P < 0.05). CONCLUSIONS: Glutathione and amifostine show a beneficial effect in experimental IFS- and ACR-induced HC. Thus, they should be investigated as an alternative treatment to prevent HC observed in patients undergoing IFS treatment. PMID- 16708235 TI - Gene-associated single nucleotide polymorphism discovery in perennial ryegrass (Lolium perenne L.). AB - Molecular genetic marker development in perennial ryegrass has largely been dependent on anonymous sequence variation. The availability of a large-scale EST resource permits the development of functionally-associated genetic markers based on SNP variation in candidate genes. Genic SNP loci and associated haplotypes are suitable for implementation in molecular breeding of outbreeding forage species. Strategies for in vitro SNP discovery through amplicon cloning and sequencing have been designed and implemented. Putative SNPs were identified within and between the parents of the F(1)(NA(6) x AU(6)) genetic mapping family and were validated among progeny individuals. Proof-of-concept for the process was obtained using the drought tolerance-associated LpASRa2 gene. SNP haplotype structures were determined and correlated with predicted amino acid changes. Gene length LD was evaluated across diverse germplasm collections. A survey of SNP variation across 100 candidate genes revealed a high frequency of SNP incidence (c. 1 per 54 bp), with similar proportions in exons and introns. A proportion (c. 50%) of the validated genic SNPs were assigned to the F(1)(NA(6) x AU(6)) genetic map, showing high levels of coincidence with previously mapped RFLP loci. The perennial ryegrass SNP resource will enable genetic map integration, detailed LD studies and selection of superior allele content during varietal development. PMID- 16708239 TI - A single open sea air dive increases pulmonary artery pressure and reduces right ventricular function in professional divers. AB - After decompression from dives, bubbles are frequently observed in the right ventricular outflow tract and may lead to vascular damage, pulmonary arterial hypertension and right ventricular overload. No data exist on the effect of open sea diving on the pulmonary artery pressure (PAP). Eight professional divers performed an open sea air dive to 30 msw. Before and postdive a Doppler echocardiographic study was undertaken. Systolic pulmonary artery pressure (SPAP) was estimated from measurement of peak flow velocity of the tricuspid regurgitant jet; the ratio between pulmonary artery acceleration times (AccT) and right ventricular ejection time (RVET) was used as an estimate of the mean PAP. No evidence of either patent foramen ovale or intra-pulmonary shunt was found in any subject postdive after performing a Valsalva maneuver. SPAP increased from 25 +/- 3 to 33 +/- 2 mmHg and AccT/RVET ratio decreased from 0.44 +/- 0.04 to 0.3 +/- 0.02 20 min after the dive, respectively. Pulmonary vascular resistance increased from 1.2 +/- 0.1 to 1.4 +/- 0.1 Woods Units. Postdive right ventricle end diastolic and end-systolic volumes were increased for about 19% (P = 0.001) and 33% (P = 0.001) and right ejection fraction decreased about for 6% (P = 0.001). Cardiac output decreased from 4.8 +/- 0.9 (l min(-1)) to 4.0 +/- 0.6 at 40 min postdive due to decreases in heart rate and stroke volume. This study shows that a single open sea dive may be associated with right heart overload due to increased pressure in the pulmonary artery. PMID- 16708240 TI - Measuring plasma oxidative stress biomarkers in sport medicine. PMID- 16708237 TI - Biophysics of P2X receptors. AB - The P2X receptor is the baby brother of the ligand-gated ion channel super family. An understanding of its role in human physiology is still developing, and no one truly knows how it works to transport ions across the membrane. In this study, we review some aspects of P2X channel biophysics, concentrating on ion permeation and gating. P2X channels transport both small and large cations and anions across cell membranes in a manner that depends on both the subunit composition of the receptor and the experimental conditions. We describe the pore properties of wild-type receptors and use the altered phenotypes of mutant receptors to point the way towards a structural model of the pore. PMID- 16708241 TI - Localization of cholinergic innervation and neurturin receptors in adult mouse heart and expression of the neurturin gene. AB - Neurturin (NRTN) is a neurotrophic factor required during development for normal cholinergic innervation of the heart, but whether NRTN continues to function in the adult heart is unknown. We have therefore evaluated NRTN expression in adult mouse heart and the association of NRTN receptors with intracardiac cholinergic neurons and nerve fibers. Mapping the regional distribution and density of cholinergic nerves in mouse heart was an integral part of this goal. Analysis of RNA from adult C57BL/6 mouse hearts demonstrated NRTN expression in atrial and ventricular tissue. Virtually all neurons in the cardiac parasympathetic ganglia exhibited the cholinergic phenotype, and over 90% of these cells contained both components of the NRTN receptor, Ret tyrosine kinase and GDNF family receptor alpha2 (GFRalpha2). Cholinergic nerve fibers, identified by labeling for the high affinity choline transporter, were abundant in the sinus and atrioventricular nodes, ventricular conducting system, interatrial septum, and much of the right atrium, but less abundant in the left atrium. The right ventricular myocardium contained a low density of cholinergic nerves, which were sparse in other regions of the working ventricular myocardium. Some cholinergic nerves were also associated with coronary vessels. GFRalpha2 was present in most cholinergic nerve fibers and in Schwann cells and their processes throughout the heart. Some cholinergic nerve fibers, such as those in the sinus node, also exhibited Ret immunoreactivity. These findings provide the first detailed mapping of cholinergic nerves in mouse heart and suggest that the neurotrophic influence of NRTN on cardiac cholinergic innervation continues in mature animals. PMID- 16708242 TI - Neural response to sustained affective visual stimulation using an indirect task. AB - Event-related potentials were recorded from 30 subjects using sustained stimulation and an indirect task, two strategies which facilitate affective responses that are complete and free of cognitive interference. Stimuli were of three types: pleasant, unpleasant and neutral. A three-phase pattern was found. The first phase, an amplitude increase in response to negative stimuli higher than to neutral and pleasant stimuli, was produced at 160 ms after stimulus onset, the prefrontal cortex being the origin of this phase. The second phase, characterized by maximal amplitudes in response to positive stimuli, was produced at 400 ms, originating in the visual cortex. Finally, the third phase, another amplitude increase in response to negative stimuli, was produced at 680 ms, and its source was located in the left precentral gyrus. Present data show that the cortical response to sustained emotional visual stimulation presented within indirect tasks provides information on attention-, motivation- and motor-related biases that complement information obtained under other experimental conditions. PMID- 16708243 TI - Synergists activation pattern of the quadriceps muscle differs when performing sustained isometric contractions with different EMG biofeedback. AB - The aims of the present study were to examine (1) endurance time and (2) activation pattern of vastus lateralis (VL), vastus medialis (VM) and rectus femoris (RF) muscles during fatiguing isometric knee extensions performed with different EMG biofeedbacks. Thirteen men (27 +/- 5 year) volunteered to participate in three experimental sessions. Each session involved a submaximal isometric contraction held until failure at an EMG level corresponding to 40% maximal voluntary contraction torque (MVC), with visual EMG biofeedback provided for either (1) RF muscle (RF task), (2) VL and VM muscles (Vasti task) or (3) the sum of the VL, VM and RF muscles (Quadriceps task). EMG activity of VL, VM and RF muscles was recorded during each of the three tasks and further analyzed. Time to task failures and MVC loss (P < 0.001) after exercises were similar (P > 0.05) between the three sessions (182 s and approximately 28%, respectively) (P > 0.05). Moreover, the magnitude of central and peripheral fatigue was not different at failure of the three tasks. Activation pattern was similar for knee extensors at the beginning of each task (P > 0.05). However, RF EMG activity decreased (P < 0.05) during the Vasti and the Quadriceps tasks (from approximately 33 to approximately 25% maximal EMG), whereas vasti EMG activity remained constant during the RF task ( approximately 41% maximal EMG). These findings suggest that (1) task failure occurs when sustaining a submaximal level of EMG activity for as long as possible and (2) CNS is not able to differentiate descending drive to the different heads of the quadriceps at the beginning of a sustained contraction, despite a different activation pattern for the bi articular RF muscle compared to the mono-articular vasti muscles during fatigue. PMID- 16708244 TI - The role of human extra-striate visual areas V5/MT and V2/V3 in the perception of the direction of global motion: a transcranial magnetic stimulation study. AB - Several published single case studies reveal a double dissociation between the effects of brain damage in separate extra-striate cortical visual areas on the perception of global visual motion defined by a difference in luminance (first order motion) versus motion defined by a difference in contrast (second-order motion). In particular, the medial extrastriate cortical region V2/V3 seems to be crucial for the perception of first-order motion, but not for second-order, whereas a lateral and more anterior portion of the cortex close to the temporo parieto-occipital junction (in the territory of the human motion area hV5/MT+) seems to be essential only for the perception of second-order motion. In order to test the hypothesis of a functional specialization of different visual areas for different types of motion, we applied repetitive transcranial magnetic stimulation (rTMS) unilaterally over areas V2/V3, V5/MT, or posterior parietal cortex (PPC) while subjects performed a 2AFC task with first- or second-order global motion displays in the contralateral visual field. Results showed a comparable disruption of the two types of motion, with both rTMS over V2/V3 or over MT/V5, and little or no effect with rTMS over PPC. The results suggest that either the previous psychophysical results with neurological patients are incorrect (highly unlikely) or that the lateral and medial regions are directly connected (as they are in macaque monkeys) such that stimulating one automatically affects the other, in this instance disruptively. PMID- 16708246 TI - Detection and estrogen regulation of leptin receptor expression in rat dorsal root ganglion. AB - Leptin receptor (OB-R) is a polypeptide consisting of a single transmembrane spanning component. OB-R widely distributes in various tissues, including the peripheral nervous system (PNS). However, there is no data about the expression of OB-R in the dorsal root ganglion (DRG). In the present study, we first detected the expressions of OB-R protein and mRNA in the rat DRG using the methods of immunohistochemistry, western blot and reverse transcriptase polymerase chain reaction (RT-PCR). Estrogen is known to influence different functions on the DRG. In this study, we observed that 17beta-estradiol (E(2)) can increase the expressions of OB-R protein and mRNA (P<0.05) in ovariectomized rat DRG and these actions can be prevented by tamoxifen which is a specific estrogen receptors (ERs) antagonist. In addition, the results of dual labeling of OB-R with the two ER isoforms, ERalpha and ERbeta indicate that 100% colocalization of ERalpha with OB-R and about 15% colocalization of ERbeta with OB-R in DRG neurons. These results indicate that OB-R is expressed in the rat DRG and E(2) may up-regulate the expression of the OB-R protein and mRNA in an ERalpha dependent way. PMID- 16708245 TI - Differential expression of spleen tyrosine kinase Syk isoforms in tissues: Effects of the microbial flora. AB - Spleen tyrosine kinase (Syk) is expressed widely in hematopoietic and non hematopoietic cells. The widespread distribution of Syk and its involvement in host defense and allergic reactions, prompted us analyze the influence of microbial exposure on Syk expression. We compared the distribution of Syk in various tissues of germ-free and conventional mice using immunohistochemistry, Western blot analysis and real time RT-PCR. Total Syk expression was similar between germ-free and conventional mice. Since it has been claimed that Syk isoforms are differentially expressed, we studied the distribution and abundance of Syk (L) and Syk (S) isoforms in tissues from these mice. In contrast to previous reports, we found broad tissue expression of Syk (S). Interestingly, in germ-free mice the amount of Syk (S) but not Syk L protein was selectively increased in lung and spleen. In summary, our study reveals new and broad tissue expression of both Syk isoforms and demonstrates that lack of microbial flora results in selectively increased expression of Syk (S) isoform in lung and spleen. PMID- 16708248 TI - [The Gini coefficient. A numerical grading for the degree of standardization of surgical subspecialities]. AB - BACKGROUND AND GOAL: General hospitals are under pressure especially under payment by DRGs (diagnosis related groups), therefore, a general trend for specialized surgical centers is obvious. For this reason knowledge of the level of standardization is important and allows strategical management of surgical subspecialties. MATERIALS AND METHODS: The Gini coefficient (GC, range: 0-1) is a measurement of the level of standardization when an economical ABC analysis is performed. Low GC values are typical for low levels of standardization. All surgical cases coded by ICD-9-CM in 1 of the 8 central operation rooms in a Swiss general teaching hospital over a period of 30 months were included. The database was analyzed by the ABC method. Due to the ABC analysis the GC for each speciality was specified. RESULTS AND DISCUSSIONS: In data period 1 a total of 11,573 and in data period 2 3,449 operations done by 7 specialities were evaluated. The GC for the Pareto distribution was 0.86. Neurosurgery had a value of only 0.34, which means that nearly every operation was unique. The highest level of standardization was reached by vascular surgery with a GC of 0.68. Compared with other industries the degree of standardization in our general hospital is low as measured by the GC. CONCLUSIONS: The GC is a valuable and objective measurement of the different procedures and standardization of surgery of given surgical subspecialties. It can be calculated from the operation room management (ORM) database and allows an objective analysis of the variety of surgical procedures per subspecialty. In this study the GC was implicated in an ORM setting for the first time. PMID- 16708249 TI - Prophylactic antibiotics in pediatric shunt surgery. AB - INTRODUCTION: The optimal antibiotic prophylaxis for pediatric shunt-related procedures is not clear. There is much inconsistency among different medical centers. This paper summarizes and analyzes the various prophylactic antibiotic regiments used for shunt-related surgeries at different pediatric neurosurgery centers in the world. MATERIALS AND METHODS: A survey questionnaire was distributed through the Pediatric Neurosurgery list-server (an e-mail-based special interest group in pediatric neurosurgery). Forty-five completed questionnaires were received, one per medical center, primarily from pediatric neurosurgeons with the following geographic breakdown: 25 from North America, 13 from Europe, and 7 from Asia and other countries. All centers routinely administered prophylactic antibiotics for shunt-related procedures. The drugs of choice were first-generation cephalosporins (23), second-generation cephalosporins (10), naficillin/oxacillin (4), vancomycin (3), clindamycin (1), amoxicillin (1), and mixed protocols in three centers. The initial drug administration ("first dose") was: in the department before transfer to operating room (5), upon arrival to operating room (11), at induction of anesthesia (13), and at initial skin incision (16). The duration of antibiotic dosage also varied: single dose (13), 24-h administration (26), 48-h administration (2), and longer than 48 h in four centers. RESULTS AND DISCUSSION: Two general tendencies were noted, common to the majority of participating centers. There was a general trend to modify antibiotic treatment protocol in "high-risk" populations. The second common theme noted in more than half of responding centers was the use of long term antibiotic treatment for externalized devices (such as externalized shunts, external ventricular drains or lumbar drains), usually till the device was in place. PMID- 16708250 TI - Closure of large myelomeningocoele with Ramirez technique. AB - INTRODUCTION: The authors present their 7-year experience in operative closure of large open myelomeningocoele with a technique introduced by Ramirez et al. in 1986. It consists of medial advancement of bilateral bipedicled musculocutanous flap based on the latissimus dorsi and maximus gluteus without any relaxing incisions or skin grafting. RESULTS AND DISCUSSION: Thirty-five babies were operated on according to this technique on first day of life. In 16 cases, large thoracic or thoracolumbosacral defect was noted, while in further 19 newborns, lumbosacral or lumbar defect occurred. The dimensions of the entire defect ranged from 4.8x5 to 8x10.8 cm. In seven babies, the affected area had more than 50 cm(2). In all patients, three-layer tension-free closure of the dorsal wound was performed in one stage. The mean operative time was 115 min (90-195 min). All but two wounds healed uneventfully. In two babies with wide lumbosacral defects, minor skin necrosis with wound dehiscence was noted which subsequently required conservative treatment. Follow-up period ranges from 6 months to 7 years. In none of the 32 children was any late complication of the reconstructive procedure noted. CONCLUSION: Ramirez technique is a very reliable method of closure of myelomeningocoele regardless of the size of the defect and its topography. It may find a wide application in babies affected by neural tube defect. PMID- 16708251 TI - Contribution of NMR spectroscopy to the differential diagnosis of a recurrent cranial mass 7 years after irradiation for a pediatric ependymoma. AB - CASE REPORT: We describe the case of a 5-year-old-boy who underwent surgery and focal radiotherapy for an anaplastic ependymoma of the fourth ventricle. One year later, a spinal metastasis was treated the same way. Six years later, a 16-mm lesion was found on a control MRI in the posterior fossa. To help the differential diagnosis between a relapse, a radio-induced modification, and a new tumor, magnetic resonance spectroscopy was performed. The main findings were a peak at the expected resonance frequency of reduced glutathione, a prominent peak of glutamate/glutamine compounds, a low N-acetylaspartate, and the absence of elevated choline. These findings were suggestive of a meningioma, although the latency between irradiation and development of the lesion was quite short. The diagnosis was confirmed by the pathological examination. CONCLUSION: This case exemplifies the fact that magnetic resonance spectroscopy provides useful biochemical information in such a clinical setting. PMID- 16708252 TI - Expansile cranioplasty for massive occipital encephalocele. AB - OBJECTIVE: This report highlights the management of infrequently encountered massive occipital encephaloceles with herniation of large amount of apparently functional cortex into the encephalocele sac. MATERIALS AND METHODS: Two children with giant occipital encephaloceles with herniation of significant brain parenchyma into the encephalocele sac were managed with expansile cranioplasty and reconstruction of the calvarial defect with autologus bone graft harvested from the adjacent parietal region. CONCLUSIONS: Giant occipital encephaloceles with significant brain parenchyma in the encephalocele sac can be considered for preservation of the herniated parenchyma and expansile cranioplasty. PMID- 16708254 TI - Hydrocephalus in Sabuncuoglu's textbook of surgery: Cerrahiyyet' ul Haniyye. PMID- 16708253 TI - Anatomy and surgery of the infected dermal sinus of the lower spine. AB - OBJECT: Cases of infected dermal sinus are scarce and detailed surgical anatomical descriptions are hardly found in literature. The clinical, radiological, and surgical findings in four cases of an infected dermal sinus located at the lower spine are presented to elucidate the pathological anatomical configuration. CLINICAL MATERIAL: The first case showed two dermal sinuses with a parallel course extra- and intradurally, ending in a confluence of cavities connected to the conus. In this case, as well as in the fourth case, the signs and symptoms were those of meningitis. The second case presented with meningitis and a subdural empyema, while the third case presented with an intradermoid intramedullary abscess at the junction between the DS and the conus. This child probably showed signs and symptoms of conus involvement as early as during pregnancy. CONCLUSION: The anatomy of the nervous elements in this congenital anomaly is heavily disturbed, more particularly in case of infection, due to extensive arachnoidal scarring. The latter renders dissection laborious and recognition of anatomical details difficult, resulting in complete excision of a dermal sinus in less than half of the cases. Despite their variability in presentation, most cases of an infected dermal sinus show similar characteristic features. PMID- 16708255 TI - Mitochondrial DNA sequence variation in Jewish populations. AB - Sequence analysis of HVRI and HVRII mitochondrial DNA was carried out on 107 Jewish samples from Ashkenazi, Oriental, North African, and Sephardic origins. Control region sequences were assigned to a haplogroup by means of the analysis of the RFLP motif -7025 AluI or by using sequence motifs. A total of 88 different haplotypes were observed with a lower incidence of unique haplotypes (68.2%) than in other populations. Four individuals with one position of sequence heteroplasmy at nucleotides 16093, 16134, 16169, and 235, respectively, were detected. The mean pairwise difference in the Jewish population was 9.7 nucleotides. The gene diversity was 0.996, and the random match probability was 1.3%. When the data were compared with the autosomal and Y-chromosome markers previously studied in these populations, sex-specific differences could be observed in the Jewish populations. This fact must be taken into account for choosing suitable databases to correctly weigh the value of the evidence of a mtDNA and/or Y profile match. PMID- 16708257 TI - Electrical excitation of the acoustically sensitive auditory nerve: single-fiber responses to electric pulse trains. AB - Nearly all studies on auditory-nerve responses to electric stimuli have been conducted using chemically deafened animals so as to more realistically model the implanted human ear that has typically been profoundly deaf. However, clinical criteria for implantation have recently been relaxed. Ears with "residual" acoustic sensitivity are now being implanted, calling for the systematic evaluation of auditory-nerve responses to electric stimuli as well as combined electric and acoustic stimuli in acoustically sensitive ears. This article presents a systematic investigation of single-fiber responses to electric stimuli in acoustically sensitive ears. Responses to 250 pulse/s electric pulse trains were collected from 18 cats. Properties such as threshold, dynamic range, and jitter were found to differ from those of deaf ears. Other types of fiber activity observed in acoustically sensitive ears (i.e., spontaneous activity and electrophonic responses) were found to alter the temporal coding of electric stimuli. The electrophonic response, which was shown to greatly change the information encoded by spike intervals, also exhibited fast adaptation relative to that observed in the "direct" response to electric stimuli. More complex responses, such as "buildup" (increased responsiveness to successive pulses) and "bursting" (alternating periods of responsiveness and unresponsiveness) were observed. Our findings suggest that bursting is a response unique to sustained electric stimulation in ears with functional hair cells. PMID- 16708260 TI - Elongation of outer transmembrane domain alters function of miniature K+ channel Kcv. AB - The virus-coded channel Kcv has the typical structure of a two-transmembrane domain K(+) channel. Exceptional are its cytoplasmic domains: the C terminus basically ends inside the membrane and, hence, precludes the formation of a cytoplasmic gate by the so-called bundle crossing; the cytoplasmic N terminus is composed of only 12 amino acids. According to structural predictions, it is positioned in the membrane/aqueous interface and connected via a proline kink to the outer transmembrane domain (TM1). Here, we show that this proline kink affects channel function by determining the position of TM1 in the membrane bilayer. Extension of the hydrophobic length of TM1 by either eliminating the proline kink or introducing an alanine in TM1 augments a time- and voltage dependent inward rectification of the channel. This suggests that the positional information of TM1 in the bilayer is transmitted to a channel gate, which is not identical with the cytoplasmic bundle crossing. PMID- 16708262 TI - A multivariate analysis of soil yeasts isolated from a latitudinal gradient. AB - Yeast isolates from soil samples collected from a latitudinal gradient (>77 degrees S to >64 degrees N) were subjected to multivariate analysis to produce a statistical foundation for observed relationships between habitat characteristics and the distribution of yeast taxa (at various systematic levels) in soil microbial communities. Combinations of temperature, rainfall (highly correlated with net primary productivity), and electrical conductivity (EC) could explain up to ca. 44% of the distribution of the predominant yeast species, rainfall and pH could explain ca. 32% of the distribution of clades in the most common orders (Filobasidiales and Tremellales), whereas vegetation type (trees, forbs, and grass) played the same role for orders. Cryptococcus species with appropriate maximum temperatures for growth predominated in most soils. Cryptococcus species in the Albidus clade of the Filobasidiales predominated in desert soils; Cryptococcus species of other clades in the Filobasidiales and Tremellales predominated in wetter and more-vegetated soils, with Tremellalean species favored in soils of lower pH or higher EC. The predominance of Cryptococcus species in soils has been attributed to their polysaccharide capsules, particularly important when competing with bacteria in arid soils. PMID- 16708264 TI - Prevention of hypopharyngeal stenosis with silastic sheeting following transoral resection. AB - Hypopharyngeal stenosis following transoral laser microresection (TLM) of hypopharyngeal carcinomas is a rare but devastating complication. Early oral intake and insertion of a nasogastric feeding tube have been thought to prevent stenosis after surgery. However, though rare, severe dysphagia can be caused by hypopharyngeal stenosis following TLM despite preventive efforts. We suggest a new way to prevent hypopharyngeal stenosis by using silastic sheeting with a feeding tube. This stent was inserted in three patients who underwent extensive TLM of hypopharyngeal carcinomas. This technique will help in the treatment and prevention of hypopharyngeal stenosis following TLM in selected patients with wide or circumferential hypopharyngeal tumors. PMID- 16708263 TI - Changes in communities of Fusarium and arbuscular mycorrhizal fungi as related to different asparagus cultural factors. AB - Asparagus (Asparagus officinalis) is a high-value perennial vegetable crop that has shown a marked decline in productivity after many years of continuous harvesting. This decline is caused by an increase in both abiotic (autotoxicity, harvesting pressure) and biotic stresses [fungal infections, mainly Fusarium crown and root rot (FCRR)]. To gain insight into disease development and possible mitigation strategies, we studied the effects of harvesting, time in the growing season, and field age on FCRR development, Fusarium species composition, and arbuscular mycorrhizal fungi (AMF) communities in both a controlled field experiment and an ecological survey of commercial fields. In one experiment, a 3 year-old asparagus field was subdivided into plots that were harvested or not and sampled throughout the growing season to assess short-term dominant Fusarium species shifts. In addition, diseased and healthy asparagus plants sampled from six commercial fields in the same geographical region were used to assess Fusarium and AMF communities in relation to different parameters. Fusarium and AMF communities were described by using a polymerase chain reaction (PCR) denaturing gradient gel electrophoresis (DGGE) approach, and results were analyzed by mainly correspondence analysis and canonical correspondence analysis. Results showed that dominant Fusarium taxa assemblages changed throughout the growing season. Harvested plots had significantly more FCRR symptomatic plants at the end of the growing season, but this effect was not related with any trend in Fusarium community structure. Sampling site and plant age significantly influenced AMF community structure, whereas only sampling site consistently influenced the Fusarium community. Diseased and healthy plants harbored similar Fusarium and AMF communities. Shifts in Fusarium community might not be responsible for different disease incidence because they are ubiquitous regardless of plant health status or harvesting regime. The different incidence noted might rather be related to plant physiology, antagonist microbial communities, or soil parameters. PMID- 16708266 TI - Recovery from mild traumatic brain injury: a focus on fatigue. AB - BACKGROUND: Fatigue is one of the most frequently reported symptoms after Mild Traumatic Brain Injury (MTBI). To date, systematic and comparative studies on fatigue after MTBI are scarce, and knowledge on causal mechanisms is lacking. OBJECTIVES: To determine the severity of fatigue six months after MTBI and its relation to outcome. Furthermore, to test whether injury indices, such as Glasgow Coma Scale scores, are related to higher levels of fatigue. METHODS: Postal questionnaires were sent to a consecutive group of patients with an MTBI and a minor-injury control group, aged 18-60, six months after injury. Fatigue severity was measured with the Checklist Individual Strength. Postconcussional symptoms and limitations in daily functioning were assessed using the Rivermead Post Concussion Questionnaire and the SF-36. RESULTS: A total of 299 out of 618 eligible (response rate 52%) MTBI patients and 287 out of 482 eligible (response rate 60%) minor-injury patients returned the questionnaire. Ninety-five MTBI patients (32%) and 35 control patients (12%) were severely fatigued. Severe fatigue was highly associated with the experience of other symptoms, limitations in physical and social functioning, and fatigue related problems like reduced activity. Of various trauma severity indices, nausea and headache experienced on the ED were significantly related to higher levels of fatigue at six months. CONCLUSIONS: In conclusion, one third of a large sample of MTBI patients experiences severe fatigue six months after injury, and this experience is associated with limitations in daily functioning. Our finding that acute symptoms and mechanism of injury rather than injury severity indices appear to be related to higher levels of fatigue warrants further investigation. PMID- 16708267 TI - Stability, controllability, and observability of the "four state" model for the sarcomeric control of contraction. AB - A model of the sarcomeric control of contraction at various loading conditions has to maintain three cardinal features: stability, controllability (where the output can be controlled by the input), and observability (where the output reflects the effects of all the state variables). The suggested model of the sarcomere couples calcium kinetics with cross-bridge (XB) cycling and comprises two feedback mechanisms: (i) the cooperativity, whereby the number of force generating (strong) XBs determines calcium affinity, regulates XB recruitment, and (ii) the mechanical feedback, whereby shortening velocity determines XBs cycling rate, controls the XBs contractile efficiency. The sarcomere is described by a set of four first-order nonlinear differential equations, utilizing the Matlab's Simulink software. Small oscillatory input was imposed when the state variables trajectories reached a steady state. The linearized state-space representations of the model were calculated for various initial sarcomere lengths. The analysis of the state-space representation validates the controllability and observability of the model. The model has four poles: three at the left side of the complex plane and one integrating pole at the origin. Therefore, the system is marginally stable. The Laplace transform confirms that the state representation is minimal and is therefore observable and controllable. The extension of the model to a multi-sarcomere lattice was explored, and the effects of inhomogeneity and nonuniform activation were described. PMID- 16708268 TI - Single particle tracking across sequences of microscopical images: application to platelet adhesion under flow. AB - A versatile and automated image processing technique and data extraction procedure from videomicroscopic data is presented. The motivation is a detailed quantification of blood platelet adhesion from laminar flow onto a surface. The characteristics of the system under observation (type of cells, their speed of movement, and the quality of the optical image to analyze) provided the criteria for developing a new procedure enabling tracking for long image sequences. Specific features of the novel method include: automatic segmentation methodology which removes operator bias; platelet recognition across the series of images based on a probability density function (two-dimensional, Gaussian-like) tailored to the physics of platelet motion on the surface; options to automatically tune the procedure parameters to explore different applications; integrated analysis of the results (platelet trajectories) to obtain relevant information, such as deposition and removal rates, displacement distributions, pause times and rolling velocities. Synthetic images, providing known reference conditions, are used to test the method. The algorithm operation is illustrated by application to images obtained by fluorescence microscopy of the interaction between platelets and von Willebrand factor-coated surfaces in parallel-plate flow chambers. Potentials and limits are discussed, together with evaluation of errors resulting from an inaccurate tracking. PMID- 16708269 TI - Combined transfer function analysis and modelling of cerebral autoregulation. AB - The clinical importance of cerebral autoregulation has resulted in a significant body of literature that attempts both to model the underlying physiological processes and to estimate the mathematical relationships between clinically measurable variables, the most common of which are Arterial Blood Pressure and Cerebral Blood Flow Velocity. These approaches have, however, rarely been used together to interpret clinical data. A simple model of cerebral autoregulation is thus proposed here, based on a flow dependent feedback mechanism with gain and time constant that adjusts arterial compliance. Analysis of this model shows that it closely approximates a second order system for typical values of physiological parameters. The model parameters can be optimally estimated from available experimental data for the Impulse Response (IR), yielding physiologically reasonable values, although there is one free parameter that must be fixed. The effects of changes in feedback gain and time constant are found to be significant on the predicted IR and can thus be estimated robustly from experimental data. The effects of elevated baseline Intracranial Pressure (ICP) are found to be exactly equivalent to a reduced feedback gain, although the solution is much less sensitive to the former effect. A transfer function approach can be used to estimate autoregulation status clinically using a physiologically-based model, thus providing greater insight into the processes that govern cerebral autoregulation. PMID- 16708270 TI - Analyzing trends in brain interface technology: a method to compare studies. AB - Continued progress in the field of Brain Interface (BI) research has encouraged the rapid expansion of the BI community over the last two decades. As the number of BI researchers and organizations steadily increases, newer and more advanced technologies are constantly produced, evaluated, and reported. Though the BI community is committed to accurate and objective evaluation of methods, systems, and technology, the diversity of the field has hindered the development of objective methods of comparison. This paper introduces a new method for directly comparing studies of BI technology based on the theoretical models and taxonomy proposed by Mason, Moore, and Birch. The effectiveness of the proposed method was demonstrated by interpreting and comparing a representative set of 21 BI studies. The method allowed us to 1) identify the salient aspects of a specific BI study, 2) identify what has been reported and what has been omitted, 3) facilitate a complete and objective comparison with other studies, and 4) characterize overall trends, areas of inactivity, and reporting practices. PMID- 16708271 TI - Role of endosomal trafficking dynamics on the regulation of hepatic insulin receptor activity: models for Fao cells. AB - Evidence indicates that endosomal insulin receptor (IR) trafficking plays a role in regulating insulin signal transduction. To evaluate its importance, we developed a series of biokinetic models for quantifying activated surface and endosomal IR dynamics from published experimental data. Starting with a published two-compartment Fao hepatoma model, a four-pool model was formulated that depicts IR autophosphorylation after receptor binding, IR endosomal internalization/trafficking, insulin dissociation from and dephosphorylation of internalized IR, and recycling of unliganded, dephosphorylated IR to the plasma membrane. Quantification required three additional data sets, two measured, but unmodeled by the same group. A five-pool model created to include endosomal trafficking of the nonphosphorylated insulin-IR complex was fitted using the same data sets, augmented with another published data set. Creation of a six-pool model added the physiologically relevant dissociation of insulin ligand from the activated endosomal IR. More importantly, all three models, validated against additional data not used in model fitting, predict that, mechanistically, internalization of activated IR is a rate-limiting step, at least under the receptor saturating conditions of the fitting data. This rate includes the transit time to a site where insulin dissociation from and/or dephosphorylation of the IR occurs by docking with protein-tyrosine phosphatases (PTPases), or where a sufficient conformational change occurs in the IR, perhaps due to insulin IR dissociation, where associated PTPases may complete IR dephosphorylation. Our new models indicate that key events in endosomal IR trafficking have significance in mediating IR activity, possibly serving to regulate insulin signal transduction. PMID- 16708272 TI - An in-vitro test set-up for evaluation of a voice-producing element under physiologic acoustic conditions. AB - To improve the voice quality of laryngectomised patients, a voice-producing element has been developed. Prior to in vivo testing we constructed and validated an in-vitro test set-up, consisting of a physical model of the subglottal tract and three physical models of the vocal tract, for the vowels /a/, /i/ and /u/ to evaluate the voice-producing element under physiologic acoustic conditions. To meet acoustic conditions described in the literature, we determined the appropriate dimensions of these physical models, using a numerical model of the pressure perturbation in rigid tubes. The numerical model showed that an acoustic equivalent of the subglottal tract could be obtained with a three-tube system and an end impedance. Vocal tract models could be constructed using two- and four resonator tubes. The physical models were built and successfully validated according to the human acoustic properties. The developed in-vitro set-up can now be applied to test voice-producing elements or vocal fold models under physiologic acoustic conditions. PMID- 16708274 TI - New York City injection drug users' memories of syringe-sharing patterns and changes during the peak of the HIV/AIDS epidemic. AB - In this oral history, 23 injection drug users (IDUs) were interviewed about the mid-1970s to mid-1980s when they could not legally purchase or possess syringes, and the threat of AIDS began to loom large. Several themes emerged, including: abrupt changes in syringe-sharing patterns; the effects of illnesses or deaths of others on their understanding of AIDS; and, racial/ethnic differences in responses to the threat of AIDS. Settings, such as "shooting galleries," helped HIV spread rapidly in the earliest stages of the city's AIDS epidemic. HIV entered the drug scene in the mid-1970s, just when IDUs were shifting from sharing homemade "works" (consisting of steel needles and syringes devised from rubber baby pacifiers and similar sources) among many IDUs to mass produced and distributed plastic, disposable needle and syringe sets. IDUs remember when they first became aware of AIDS and began to adjust their behaviors and social assumptions. PMID- 16708275 TI - Child-directed interaction: prediction of change in impaired mother-child functioning. AB - The first phase of parent-child interaction therapy (PCIT), called child-directed interaction, teaches parents to use positive and differential social attention to improve the parent-child relationship. This study examined predictors of change in mother and child functioning during the child-directed interaction for 100 mother-child dyads. The children were 3-6-years-old and diagnosed with oppositional defiant disorder. After establishing that significant improvements occurred in mother report of child disruptive behavior, parenting stress, and parenting practices, these three variables were combined to form a latent impaired mother-child functioning construct. Structural equation models were examined using maternal demographic and psychosocial variables as predictors of impaired mother-child functioning before and after the child-directed interaction. Mothers' self-reported daily hassles and depressive symptomatology predicted 74% of variance in impaired mother-child functioning before treatment. Mothers' report of social support predicted impaired mother-child functioning after the child-directed interaction, with 57% of the variance accounted for in this longitudinal model. These findings suggest the importance of improving maternal social support during the initial phase of PCIT. PMID- 16708276 TI - Symptom increase following a functional capacity evaluation in patients with chronic low back pain: an explorative study of safety. AB - INTRODUCTION: This study was performed to study intensity and duration of symptom increase following an FCE and to explore safety of an FCE. METHODS: Included were 92 patients with chronic low back pain (CLBP), mean age 38.5 years, mean self reported disability 12.5 (Roland Morris Disability Questionnaire). All patients underwent an FCE. Symptom increase was measured with a 2-item questionnaire. Operational definition for safety: no formal complaint filed and symptom increase to occur only temporarily. RESULTS: No formal complaints were filed (n=92). In total, 54 patients returned the questionnaire (59%; 'responders'). Of the responders, 76% reported increased symptom intensity after an FCE, ranging from 'little increase' to 'severe increase'. Symptoms of all responders returned to pre-FCE level. Duration of symptom increase of the responders ranged from 1 day to 3 weeks. Symptom increase resided to pre-FCE level within 1 week in 93% of the responders. Symptom increase was weakly related to self-reported disability (r=0.38, p<0.05). Except for gender, differences between responders and non responders were non-significant. CONCLUSION: A temporary increase in symptom intensity following an FCE is common. Within the operational definitions of safety used in this study, assessment of functional capacity of patients with CLBP appears safe. PMID- 16708277 TI - Factors predicting work ability following multidisciplinary rehabilitation for chronic musculoskeletal pain. AB - INTRODUCTION: This study aimed to investigate the outcome and outcome predictors of multidisciplinary rehabilitation in terms of working ability. METHODS: One hundred and forty three (n=143) patients with musculoskeletal pain (mean age=45.7, SD=8.9) were included. Work status, pain, functional health status and psychosocial factors were collected previous to treatment, after a 5 week intensive training and a 52 week follow-up period. Demographics and data on personal characteristics were also collected. RESULTS: Workability increased from 57.4 to 80% during treatment period. Stepwise multivariate logistic regression indicated that age, sleeplessness, cognitive function, overall health, pain experience, and anxiety were the strongest predictors of work ability. Pain severity and depression were not found to be significant predictors of work ability. CONCLUSIONS: These data suggest that emotional distress, cognitive function and overall health are important priority areas in rehabilitation programmes to improve work ability. PMID- 16708278 TI - Tumor microvascular architecture phenotype (T-MAP) as a new concept for studies of angiogenesis and oncology. AB - Heterogeneities exist in endothelial cells and microvascular architecture during tumor angiogenesis. We found significantly variable expression profiles of EC markers, including ephrinB2 and its receptor EphB4, and various types of the architecture. We propose a new concept of tumor microvascular architecture phenotype (T-MAP), reflecting the density, morphology, structure and the three dimensional distribution of newly formed vessels. The pattern of T-MAP may represent the invasiveness of the tumor therefore predict the outcome of therapy. The presence of T-MAP heterogeneity (T-MAPH) may be utilized as additional diagnostic criteria and for therapeutic designs for antiangiogenesis. PMID- 16708280 TI - Valve movement response of the mussel mytilus galloprovincialis to metals (Cu, Hg, Cd and Zn) and phosphate industry effluents from Moroccan Atlantic coast. AB - Valve activity was measured in the Mediterranean mussel Mytilus galloprovincialis in response to sublethal concentrations of four metals (Hg, Cu, Zn and Cd) and two phosphate industry effluents from the Atlantic coast of Morocco. Valve movements were monitored using a proximity inductive sensor which could display all activity figures from full closure to wide opening of the shell valves. In a 1 h exposure experiments, all metals induced a decrease in the time of normal opening and the appearance of sequences of stress behaviour, including enhanced valve adductions and complete closure at high concentrations. Mercury (tested from 5 to 75 microg Hg l(-1)) was the most toxic to the valve activity, with a threshold effective concentration at 10 microg Hg l(-1) and full valve closure occurring at 50 microg Hg l(-1). Copper (15-150 microg Cu l(-1)) showed a toxic effect starting at threshold concentration of 20 microg Cu l(-1) and induced full valve closure at 150 microg Cu l(-1). Zinc (100-500 microg Zn l(-1)) was effective in reducing the time of normal opening (threshold concentration at 100 microg Zn I(-1)) but no complete closure was recorded in any of the tested concentrations. For cadmium (1000-5000 microg Cd l(-1)), the valve activity was insensitive for exposures under 2000 microg Cd l(-1). Results for the testing of several samplings of the phosphate industry effluents (Safi and Jorf Lasfar) showed that their toxicity varied over the time. The effluent of the Jorf Lasfar plant (2-9.4%) was, however, more toxic than that of Safi (1-25%). In the light of these results, the sensitivity of the valve activity of Mytilus galloprovincialis to pollutants and its usefulness for in situ monitoring of coastal pollution in Morocco are discussed. PMID- 16708281 TI - Assessment of biochemical components and enzyme activities in the estuarine crab Scylla tranquebarica from naphthalene contaminated habitants. AB - To document the impact of naphthalene, a comparative toxicological research was performed in the estuarine crab Scylla tranquebarica habitant of Ennore creek (polluted site) and Kovalam creek (pristine site) for a period of 6 months. The biochemical constituents, such as protein, carbohydrate, lipid and enzyme activities like acid and alkaline phosphatase, aspartate and alanine transaminase were analysed in hepatopancreas, heamolymph and ovary of the female crabs collected from the two creeks. The results revealed that there was a significant measurable difference in these parameters in the tissues of crabs sampled from the polyaromatic hydrocarbon (PAH) polluted site (Ennore creek) when compared with the reference site (Kovalam creek). In addition naphthalene was detected in gill, hepatopancrease, heamolymph and ovary of the crabs sampled from the polluted creek. The results indicate that the indicators selected for toxic impact in S. tranquebarica may be related to the uptake of naphthalene in the tissues examined and support the feasibility of employing these types of analyses as biochemical biomarkers for PAH contaminant-exposed organisms. PMID- 16708282 TI - Biodegradability of tetrabromobisphenol A and tribromophenol by activated sludge. AB - Laboratory reactor systems based on the conventional (aerobic) activated sludge process and on the contact (anaerobic) process were operated almost one year in order to develop a biological process for the degradation of the fire retardant tetrabromobisphenol A (TBBPA) and to find out if its degradation might result in the formation of the endocrine disruptor bisphenol A (BPA). The reactors were fed a TBBPA waste mixture containing also tribromophenol (TBP), and added with contaminated sediments that might have contained indigenous bacteria exposed to these compounds. Various organic compounds were used as potential electron donors to enhance growth of halorespiring bacteria that would debrominate the TBBPA and make it available for further aerobic mineralization. In spite of the various operating strategies applied and the different carbon sources added, no TBBPA biodegradation has been observed and no accumulation of intermediates such as BPA in any of the aerobic or anaerobic reactors has been detected. TBP on the other hand, was found to be easily biodegraded by aerobic cultures simulating the activated sludge process. This was linked to consistent accumulation of bromides, released to the liquid following TBP breakdown. PMID- 16708283 TI - Associations among gender-linked toy preferences, spatial ability, and digit ratio: evidence from eye-tracking analysis. AB - Eye-tracking technology was used to monitor eye-movements in 64 adults (age range, 18-22 years) during simultaneous presentation of "masculine" and "feminine" toys. Women and men who showed more visual fixations on male-typical toys compared to female-typical toys had significantly better targeting ability and smaller (i.e., more masculine) digit ratios, a putative marker of prenatal androgen levels. In contrast, individuals with visual preferences for female typical or male-typical toys did not differ in mental rotations ability and in their retrospective reports of childhood gender-linked activities. The finding that targeting ability and digit ratios varied according to visual preferences for gender-linked toys suggests that prenatal androgens promote enduring preferences for male-typical objects and indicate further that some gender-linked traits vary according to the direction of a visual preference for gender-linked toys. Visual preferences derived from eye-tracking, therefore, may be a useful supplement to current measures of psychosexual differentiation in hormone behavior research, particularly because eye-movements are not dependent on verbal abilities or subjective evaluations of behavior. PMID- 16708284 TI - Grade failure and special education placement in sexual offenders' educational histories. AB - A sample of 701 adult men underwent assessment following illegal or clinically significant sexual behaviors or interests. Patients were categorized on the basis of phallometric (penile) responses in the laboratory to erotic stimuli depicting adults, pubescent children, and prepubescent children; histories of sexual offenses; and self-reported sexual interests. Comprising the categories were men sexually interested in prepubescent children (pedophiles; n = 114), men sexually interested in pubescent children (hebephiles; n = 377), men sexually interested in adults and who had committed a sexual offense against an adult (teleiophilic offenders; n = 139), and men sexually interested in adults and who had no known history of any sexual offenses (teleiophilic nonoffenders; n = 71). Patients' assessments included IQ testing and self-reported academic history, which included any grade failures and assignment to special education classes. Relative to the teleiophilic offenders, both the pedophilic and the hebephilic groups showed approximately double the odds of failing a grade or being enrolled in special education, both before and after covarying IQ. No significant differences were detected between the teleiophilic offenders and the teleiophilic nonoffenders. These data are consistent with the hypothesis that an erotic age preference for children sometimes results from a perturbation of neurodevelopment occurring early in life. PMID- 16708286 TI - Viral transduction of cocaine hydrolase in brain reward centers. AB - 1. Site-directed mutagenesis of human plasma butyrylcholinesterase has led to novel hydrolases that rapidly destroy cocaine. We are investigating whether viral gene transfer of such enzymes might reduce addiction liability by blocking cocaine from its sites of action. 2. As groundwork for a possible gene therapy, we previously studied adenoviral transduction of cocaine hydrolases in the rat. Systemically injected vectors raised plasma cocaine hydrolase activity greatly, reduced pressor responses to cocaine, and lowered cocaine's tissue burden. 3. In the present study, to reduce cocaine's brain access still further, vectors were injected directly into the nucleus accumbens. Six days later, medium sized neurons gained dramatic butyrylcholinesterase activity. Species-selective immunohistochemistry proved that the transgene accounted for this activity. 4. Since the transgene product is so efficient with cocaine as a substrate, it is now reasonable to begin testing gene therapy in rodent models of cocaine addiction. PMID- 16708285 TI - Transcriptional regulation of neurogenesis in the olfactory epithelium. AB - 1. The olfactory epithelium (OE) is a simple structure that gives rise to olfactory sensory neurons (OSNs) throughout life. 2. Numerous transcription factors (TFs) are expressed in regions of the OE which contain progenitor cells and OSNs. The function of some of these TFs in OSN development has been elucidated with the aide of transgenic knockout mice. 3. We review here the current state of knowledge on the role of TFs in OE neurogenesis and relate the expression of these TFs, where possible, to the well-documented phenotype of the cells as they progress through the OSN lineage from progenitor cells to mature neurons. PMID- 16708287 TI - The on-line study of sentence comprehension: an examination of dual task paradigms. AB - This paper presents three studies which examine the susceptibility of sentence comprehension to intrusion by extra-sentential probe words in two on-line dual task techniques commonly used to study sentence processing: the cross-modal lexical priming paradigm and the unimodal all-visual lexical priming paradigm. It provides both a general review and a direct empirical examination of the effects of task-demand in the on-line study of sentence comprehension. In all three studies, sentential materials were presented to participants together with a target probe word which constituted either a better or a worse continuation of the sentence at a point at which it was presented. Materials were identical for all three studies. The manner of presentation of the sentence materials was, however, manipulated; presentation was either visual, auditory (normal rate) or auditory (slow rate). The results demonstrate that a technique in which a visual target probe interrupts ongoing sentence processing (such as occurs in unimodal visual presentation and in very slow auditory sentence presentation) encourages the integration of the probe word into the on-going sentence. Thus, when using such 'sentence interrupting' techniques, additional care to equate probes is necessary. Importantly, however, the results provide strong evidence that the standard use of fluent cross-modality sentence investigation methods are immune from such external probe word intrusions into ongoing sentence processing and are thus accurately reflect underlying comprehension processes. PMID- 16708288 TI - Evolution of renal function and Na+, K +-ATPase expression during ischaemia reperfusion injury in rat kidney. AB - The aim of the present work was to study the effects of an unilateral ischaemic reperfusion injury on Na+, K+-ATPase activity, alpha1 and beta1 subunits protein and mRNA abundance and ATP content in cortical and medullary tissues from postischaemic and contralateral kidneys. Right renal artery was clamped for 40 min followed by 24 and 48 h of reperfusion. Postischaemic and contralateral renal function was studied cannulating the ureter of each kidney. Postischaemic kidneys after 24 (IR24) and 48 (IR48) hours of reperfusion presented a significant dysfunction. Na+, K+-ATPase alpha1 subunit abundance increased in IR24 and IR48 cortical tissue and beta1 subunit decreased in IR48. In IR24 medullary tissue, alpha1 abundance increased and returned to control values in IR48 while beta1 abundance was decreased in both periods. Forty minutes of ischaemia without reperfusion (I40) promoted an increment in alpha1 mRNA in cortex and medulla that normalised after 24 h of reperfusion. beta1 mRNA was decreased in IR24 medullas. No changes were observed in contralateral kidneys. This work provides evidences that after an ischaemic insult alpha1 and beta1 protein subunit abundance and mRNA levels are independently regulated. After ischaemic-reperfusion injury, cortical and medullary tissue showed a different pattern of response. Although ATP and Na+, K+-ATPase activity returned to control values, postischemic kidney showed an abnormal function after 48 h of reflow. PMID- 16708290 TI - Mental health services for children in large, employer-based health plans, 1999. AB - This study examines 1999 data from Medstat's MarketScan database of privately insured employees of US firms and their dependents. Of enrolled children and adolescents ages 2-18, 6.6% had claims for mental health services. Average outpatient expenditures per user were $651. Of children/adolescents with claims for mental health services (MH claimants), 3.4% had inpatient MH services, with an average length of stay of 8.9 days and average MH-related inpatient expenditure per user of $7,048. One half of MH claimants who had pharmacy benefit data had claims for psychotropic medications, with average expenditures per user of $328. Whereas children/adolescent mental health users comprised 8.3% of all service users, expenditures for their care were 20.5% of all service expenditures for children/adolescents in private health plans. Results also highlight the importance of including data on psychotropic medication in analysis of children's MH services utilization, as well as the need to consider the use of psychotropic medications among children/adolescents who do not utilize other MH services. PMID- 16708289 TI - Understanding obsessive-compulsive disorder: focus on decision making. AB - Current approaches to obsessive-compulsive disorder (OCD) have suggested that neurobiological abnormalities play a crucial role in the etiology and course of this psychiatric illness. In particular, a fronto-subcortical circuit, including the orbitofrontal cortex, basal ganglia and thalamus appears to be involved in the expression of OCD symptoms. Neuropsychological studies have also shown that patients with OCD show deficits in cognitive abilities that are strictly linked to the functioning of the frontal lobe and its related fronto-subcortical structures, such as executive functioning deficits and insufficient cognitive behavioral flexibility. This article focuses on decision making, an executive ability that plays a crucial role in many real-life situations, whereby individuals choose between pursuing strategies of action that involve only immediate reward and others based on long-term reward. Although the role of decision-making deficits in the evolution of OCD requires further research, the collected findings have significant implications for understanding the clinical and behavioral heterogeneity that characterizes individuals with OCD. PMID- 16708292 TI - Technique of pancreaticojejunostomy reconstruction after pancreaticoduodenectomy. PMID- 16708291 TI - Influx of extracellular Ca2+ involved in jasmonic-acid-induced elevation of [Ca2+]cyt and JR1 expression in Arabidopsis thaliana. AB - The changes in cytosolic Ca2+ levels play important roles in the signal transduction pathways of many environmental and developmental stimuli in plants and animals. We demonstrated that the increase in cytosolic free Ca2+ concentration ([Ca2+]cyt) of Arabidopsis thaliana leaf cells was induced by exogenous application of jasmonic acid (JA). The elevation of [Ca2+]cyt was detected within 1 min after JA treatment by the fluorescence intensity using laser scanning confocal microscopy, and the elevated level of fluorescence was maintained during measuring time. With pretreatment of nifedipine (Nif), a nonpermeable L-type channel blocker, the fluorescence of [Ca2+]cyt induced by JA was inhibited in a dose-dependent manner. In contrast, verapamil, another L-type channel blocker, had no significant effect. Furthermore, Nif repressed JA-induced gene expression of JR1 but verapamil did not. JA-induced gene expression could be mimicked by higher concentration of extracellular Ca2+. W-7 [N-(6-aminohexyl)-5 chloro-1-naphthalenesulfonamide], an antagonist of calmodulin (CaM), blocked the JA induction of JR1 expression while W-5 [N-(6-aminohexyl)-1 naphthalenesulfonamide], its inactive antagonist, had no apparent effect. These data provide the evidence that the influx of extracellular Ca2+ through Nif sensitive plasma membrane Ca2+ channel may be responsible for JA-induced elevation of [Ca2+]cyt and downstream gene expression, CaM may be also involved in JA signaling pathway. PMID- 16708293 TI - Somatostatin analogues in the prevention of pancreas-related complications after pancreatic resection. AB - BACKGROUND/PURPOSE: The Achilles' heel of operative pancreatectomies is the pancreaticoenterostomy for proximal resections and the pancreatic parenchymal closure for distal resections. Inhibition of pancreatic exocrine secretions by somatostatin analogues has been suggested to decrease pancreas-specific complications, but this topic remains controversial. METHODS: We performed a randomized, prospective, placebo-controlled, multicenter trial of the use of perioperative vapreotide, a potent somatostatin analogue, in pancreatic resections for presumed neoplasms in 381 patients without chronic pancreatitis. We also reviewed the literature on the use of somatostatin and its analogues after pancreatectomy. RESULTS: When compared to the placebo, perioperative vapreotide had no effect on overall pancreas-specific complications (30.4% vs 26.4%), mortality (0% vs 1.4%), overall complications (40% vs 42%), and duration of hospitalization; there were no differences in complications per type of resection with use of vapreotide--proximal versus distal resection. Seven other prospective, randomized trials provide differing results. CONCLUSIONS: Our study with vapreotide failed to show any benefit when administered perioperatively (and for 7 days postoperatively) on pancreas-specific complications after major pancreatectomy in patients without chronic pancreatitis. The use of perioperative analogues that suppress pancreatic exocrine secretion seems not to be warranted as routine treatment. PMID- 16708294 TI - Pancreaticojejunostomy using duct-to-mucosa anastomosis without a stenting tube. AB - BACKGROUND PURPOSE: There is a high risk of anastomotic leakage after pancreaticojejunostomy following pancreaticoduodenectomy in patients with a normal soft pancreas because of the high degree of exocrine function. Therefore, pancreaticojejunostomy is generally performed using a stenting tube (stented method). However, pancreaticojejunostomy with a certain duct-to-mucosa anastomosis does not always require a stenting tube, even in patients with a normal soft pancreas. Recently, we have performed pancreaticojejunostomy with duct-to-mucosa anastomosis without a stenting tube (nonstented method) and obtained good results. METHODS: The point of this technique is to maintain adequate patency of the anastomosis using a fine atraumatic needle and monofilament thread. The results of end-to-side pancreaticojejunostomy of the normal soft pancreas using the nonstented method (n = 123) were compared with those using the stented method (n = 45). RESULTS: There were no differences in background characteristics between the groups, including age, gender, and disease. The mean times to complete pancreaticojejunostomy were around 30 min in the two groups and the rates of morbidity and leakage of pancreaticojejunostomy were 26.8% and 5.7% in the nonstented group and 22.2% and 6.7% in the stented group, respectively. These differences were not statistically significant. One patient in the stented group died of sepsis following leakage of pancreaticojejunostomy. There were also no significant differences in the mean time to initiation of solid food intake or postoperative hospital stay. CONCLUSIONS: In conclusion, complete pancreaticojejunostomy using duct-to-mucosa anastomosis for a normal soft pancreas does not require a stenting tube. This nonstented method can be considered one of the basic procedures for pancreaticojejunostomy because of its safety and certainty. PMID- 16708295 TI - Is pancreaticogastrostomy safer than pancreaticojejunostomy? AB - BACKGROUND/PURPOSE: Failure of a pancreatic-enteric anastomosis very frequently leads to morbidity and mortality after pancreaticoduodenectomy. Pancreaticojejunostomy or pancreaticogastrostomy is often used after pancreaticoduodenectomy. The many reports on pancreaticogastrostomy support the low rates of anastomotic leakage and mortality compared with pancreaticojejunostomy. METHODS: Between January 1995 and December 2004, 155 pancreaticojejunostomies and 58 pancreaticogastrostomies were performed after pancreatic resection in the Second Department of Surgery of Nagoya University Hospital. Postoperative morbidity and mortality were analyzed. RESULTS: The incidence of pancreatic fistula was similar for the pancreaticojejunostomy (12.2%) and pancreaticogastrostomy (20.7%) groups and the mortality rate was 0% in both groups. CONCLUSIONS: This restrospective clinical study suggested no significant difference in the incidence rate of pancreatic fistula and mortality between pancreaticojejunostomy and pancreaticogastrostomy. PMID- 16708296 TI - Complications after pancreaticoduodenectomy: the problem of current definitions. AB - Pancreaticoduodenectomy nowadays represents a complex procedure and a challenge for the surgeon. Even though mortality is reported to be below 5% for experienced surgeons, morbidity is still around 30%-50%, often leading to prolongation of hospital stay, demanding postoperative investigations and procedures, and outpatient monitoring of the patients with complications. In the literature there is no agreement on the definitions of postoperative complications following pancreaticoduodenectomy, leading to a wide range of complication rates in different specialist units, particularly regarding the source of every complication, postoperative pancreatic fistula, and others such as delayed gastric emptying. Some authors have demonstrated that applying different definitions in homogeneous, single-center series, the incidence of a complication varied with statistical significance, implying the impossibility of correctly comparing different experiences. It seems essential to organize a Consensus Meeting among expert surgeons to prepare world-wide accepted definitions. The aim of this article is to review the current controversial definitions and to suggest a new clinical-based approach to the problem of the feasibility and reliability of the definitions themselves. PMID- 16708298 TI - Gastrointestinal function and quality of life after pylorus-preserving pancreatoduodenectomy. AB - The pylorus-preserving pancreatoduodenectomy (PPPD) has taken the place of the conventional Whipple pancreatoduodenectomy as the standard procedure for various periampullary disease. With recent advances in surgical techniques and improvements in perioperative management, the number of long-term survivors after PPPD is increasing. As a result, surgeons should pay more attention to the patients' postoperative gastrointestinal function, nutrition, and quality of life (QOL). Gastric stasis, which is a frequent complication during the early postoperative period after PPPD, prolongs the hospital stay and impairs the QOL in the intermediate term. Several possible pathogeneses for this gastric stasis have been postulated; however, the precise mechanism remains unclear. The gastric emptying function gradually recovers to the preoperative level by 6 months after PPPD. Pancreatic functions are likely to be maintained for at least 1 year after PPPD; however, in some cases, they tend to gradually deteriorate over time after the operation, depending on the type of pancreatic reconstruction or the preoperative condition of the pancreas. It is important to note that preoperative and postoperative pancreatic exocrine function strongly influence the postoperative outcome regarding such factors as pancreatic fistula, body weight maintenance, nutrition, and the QOL. The QOL, as assessed by questionnaire, normally returns to the preoperative level within 6 months after PPPD, and this correlates with the changes in gastrointestinal function and nutritional status. It still remains an unresolved question, however, whether the Billroth-I PPPD really leads to better long-term nutritional status, but worse early gastric emptying function, than the Billroth-II type of reconstruction. PMID- 16708297 TI - A new technique for pancreaticogastrostomy for the soft pancreas: the transfixing suture method. AB - BACKGROUND/PURPOSE: Pancreatic anastomotic leakage remains a persistent problem after pancreaticoduodenectomy (PD). The presence of soft, nonfibrotic pancreatic tissue is one of the most important risk factors for pancreatic leakage. Accordingly, we devised a pancreas-transfixing suture method for pancreaticogastrostomies in patients with a soft, nonfibrotic pancreatic remnant. METHODS: The pancreas-transfixing method was applied in 103 consecutive patients after either standard PD (49 patients) or pylorus-preserving pancreaticoduodenectomy (PPPD) (54 patients) for malignant or benign disease. Of these 103 patients, 65 had a soft, nonfibrotic pancreatic remnant. For the pancreaticogastrostomy technique, an ultrasonically activated scalpel was used for transecting the pancreas. The inner layer involves a duct-to-mucosa anastomosis with an internal stent and the outer layer involves a single row of pancreas-transfixing sutures between the pancreatic remnant and the posterior gastric wall. RESULTS: Operative mortality was zero and morbidity was 22%. Only two patients (2%) developed pancreatic leaks; both resolved nonoperatively with the continuation of closed drainage. CONCLUSIONS: This technique is simple and appears to reduce the risk of pancreatic leakage, possibly by decreasing the risk of suture injury of the pancreas and by embedding the transected stump into the wall of the stomach. This novel pancreaticogastrostomy technique is an effective reconstructive procedure, especially for patients with a soft, nonfibrotic pancreas. PMID- 16708299 TI - Safe Veress needle insertion. AB - BACKGROUND/PURPOSE: For laparoscopic surgery, the creation of pneumoperitoneum still remains a must. The insertion of a Veress needle or a trocar is never perfectly safe, and almost every kind of intraabdominal organ injury due to these insertions has been reported worldwide. Here, we describe a safe technique for creating pneumoperitoneum. METHODS: For the creation of pneumoperitoneum, under direct vision, the linea alba was elevated with two towel clips and then the Veress needle was inserted. We reviewed 368 patients operated on with this technique for complication rates. Ultrasound images were obtained before and during abdominal-wall lifting in 10 patients. RESULTS: There were no injuries due to the insertion of the Veress needle or trocars. In 90% of the patients, pneumoperitoneum was created successfully on the first attempt. Ultrasound examination demonstrated a mean extra safe area of 11.8 mm during abdominal-wall lifting with this technique. CONCLUSIONS: Elevating the linea alba during Veress needle insertion is safe. PMID- 16708300 TI - Immunohistochemical demonstration of c-Kit protooncogene product in gallbladder cancer. AB - BACKGROUND/PURPOSE: Although some gallbladder carcinomas are immunoreactive for c Kit, the reasons for the c-Kit expression and its clinicopathologic implications are unknown. METHODS: We investigated the prevalence of c-Kit immunoreactivity, its clinicopathologic correlates (including microvessel density and postoperative outcome), and the possible mechanisms of c-Kit expression. We reviewed retrospectively, the clinicopathologic records of 47 patients who had undergone macroscopically complete gallbladder carcinoma resection. The numbers of patients at pathologic stages I to IV, according to current TNM-based staging, were 10, 5, 18, and 14, respectively. For immunohistochemical examination, we used monoclonal antibodies against c-Kit and CD 34 (progenitor cell markers), cytokeratin 7 and cytokeratin 19 (cholangiocyte markers), and OCH1E5 (a hepatocyte marker). Control tissue samples were from five gallbladder specimens each with chronic cholecystitis, polyp, and adenoma. RESULTS: Cytoplasmic immunostaining for c-Kit was detected in 21 of the 47 gallbladder carcinomas (45%), and in 1 of the 15 control specimens (diagnosis, chronic cholecystitis). Young age was significantly associated with c-Kit positivity; however, there were no significant differences in the incidence of c-Kit positivity among other variables, including tumor stage and outcome. However, microvessel density was significantly higher in c-Kit positive gallbladder carcinoma compared with c-Kit-negative gallbladder carcinoma. None of the 47 cancer specimens or the 15 control specimens were stained for CD34 and OCH1E5, but all 47 cancer specimens were stained for cytokeratins 7 and 19. CONCLUSIONS: Gallbladder carcinomas positive for c-Kit are unlikely to arise from immature cells, but may be associated with neovascularization. Angiogenesis inhibitors, such as tyrosine kinase inhibitors, therefore, may suppress the growth of some gallbladder cancers. PMID- 16708301 TI - Intraductal low papillary histological pattern of carcinoma component shows intraductal spread in invasive carcinoma of the pancreas. AB - BACKGROUND/PURPOSE: We attempted to discriminate between carcinoma in situ (CIS) and the intraductal invasion/cancerization of invasive ductal carcinoma (IDC) of the pancreas, by comparing the histological patterns of the intraductal components and those of venous invasion. METHODS: Specimens from 30 patients with IDC were examined histopathologically. Intraductal components and blood vessel invasion in IDC were assessed in specimens stained with hematoxylin & eosin and elastica van Gieson (EVG). RESULTS: Intraductal components of IDC were found in 28 of the 30 cases of IDC, in 261 ducts, and in 2.3 ducts per one section of one case, on average. The intraductal components of IDC were classified into three histological patterns, as follows: low papillary (including flat), tubular (including solid and cribriform), and mixed (low papillary plus tubular). The incidences of the low papillary, tubular, and mixed patterns in the 261 ducts, were 39% (102 ducts), 56% (145 ducts), and 5% (14 ducts), respectively. The histological pattern of venous invasion was tubular in all but 1 of the 26 cases, and this 1 case showed low papillary patterns as well as a tubular pattern. CONCLUSIONS: A tubular pattern of intraductal components in IDC of the pancreas indicates intraductal invasion, while a low papillary pattern indicates CIS or carcinoma in another location to which it has spread. PMID- 16708302 TI - Clinical significance of serum cytokeratin-19 fragment (CYFRA 21-1) in hepatocellular carcinoma. AB - BACKGROUND/PURPOSE: CYFRA 21-1, a soluble fragment of cytokeratin 19, is increased in serum in some patients with hepatocellular carcinoma, but the clinical significance of this increase is still unknown. METHODS: Serum concentrations of CYFRA 21-1 were measured in 240 patients with hepatocellular carcinoma prior to hepatic resection. The relationships between serum CYFRA 21-1 concentrations and clinicopathologic features were analyzed. RESULTS: The sensitivity of CYFRA 21-1 as a test for hepatocellular carcinoma was 18.8%. Serum CYFRA 21-1 was significantly higher in patients with portal vein tumor thrombus, and serum CYFRA 21-1 increased with the progression of portal vein tumor thrombus. Tumor size was related to serum CYFRA 21-1, but there were no significant correlations between serum CYFRA 21-1 concentrations and tumor differentiation or number of tumors. Although patients with stage IV tumor had significantly higher CYFRA 21-1 concentrations than those with stages I, II, and III, CYFRA 21-1 was not associated with postoperative prognosis. CONCLUSIONS: Although high concentrations of CYFRA 21-1 were often detected in patients with a tumor diameter greater than 5 cm or tumor thrombus in the major portal vein, CYFRA 21-1 is not a useful diagnostic tool for hepatocellular carcinoma because of its low sensitivity. PMID- 16708303 TI - Clinicopathological implications of immunohistochemically demonstrated mucin core protein expression in hepatocellular carcinoma. AB - METHODS: We examined the expression of mucin core protein 1 (MUC1) immunohistochemically in 186 surgical specimens of histopathologically nonmucinous hepatocellular carcinoma (HCC) and compared the clinicopathological features in patients with MUC1-positive HCC (MUC1-positive group) with those in patients with MUC1-negative HCC (MUC1-negative group). RESULTS: MUC1 immunoreactively was present in 85 of the 186 HCCs. Of the clinicopathological variables examined, the serum concentration of alpha-fetoprotein, tumor differentiation, bile duct invasion, lymph node metastasis, and cytokeratin 19 expression exhibited significant associations with MUC1 expression. Although cumulative and tumor-free survival rates were not different between the two groups, the percentage of patients with first recurrence of HCC in distant organs (distant metastasis) within 2 years after surgery was significantly higher in the MUC1-positive group than in the MUC1-negative group (P = 0.0104). The risk ratio of MUC1 positivity for this type of distant metastasis was 3.156 (95% confidence interval, 1.064-9.358). CONCLUSIONS: In patients with MUC1-positive HCC, careful follow-up is necessary, not only for intrahepatic recurrence but also for distant metastasis, after the resection of primary HCC. PMID- 16708304 TI - Amebic liver abscess rupturing into the lesser omentum space. AB - A case of an amebic abscess localized in the lesser omentum is reported. There was no sign of a liver abscess in the imaging examination or the operative findings. However, it is likely that the amebic infection occurred after a liver abscess ruptured into the abdominal cavity. Early diagnosis and therapy are required when an abscess of unknown origin borders the liver, given the possibility of amebic abscess. PMID- 16708305 TI - Treatment of colorectal liver metastasis with biliary and portal vein tumor thrombi by hepatopancreatoduodenectomy. AB - We present a case of a large colorectal liver metastasis with portal vein and biliary tumor thrombi and duodenal and jejunal direct invasion that required hepatopancreatoduodenectomy. A 38-year-old woman presented to her local hospital with right back pain and jaundice. She had undergone transverse colectomy and limited liver resection for transverse colon cancer with a synchronous liver metastasis in September 1991, and low anterior resection for rectal carcinoma in January 1996. She was diagnosed as having colorectal liver metastasis and was referred to our hospital for possible surgery. Radiologic and endoscopic examinations revealed a large liver tumor occupying the right lobe, biliary dilation in the left lateral section, and a portal vein tumor thrombus. Invasion of the inferior vena cava and the right renal vein were also suspected. Intraoperative findings revealed a large liver tumor that occupied the right lobe and invaded the duodenum and jejunum. The tumor was resected successfully by right trisectionectomy, caudate lobectomy, pancreatoduodenectomy, partial resection of the jejunum, and combined portal vein resection and reconstruction. The inferior vena cava, right kidney, and renal vein could be detached from the tumor. The patient has enjoyed an active life without recurrence for 2 years since the operation. PMID- 16708307 TI - A mouse model of breast cancer metastasis to the choroid of the eye. AB - Transformed mouse mammary epithelial cells, r3T, injected into the arterial circulation form bone metastases with high frequency. Here we report that metastases to the choroid of the eye also occur in these mice with a penetrance of at least 50%. The tumors can occupy as much as half the volume of the eye, and pigmented cells become incorporated into and distributed throughout the tumors. Pigmentation is also observed in the brains and optic nerves of mice with choroidal tumors, suggesting that the tumor cells stimulate migration of pigmented cells along the optic nerve into the brain. To our knowledge, this is the first mouse model of breast cancer choroidal metastasis, and should be useful in the study of this disease. PMID- 16708308 TI - Adult outcome of congenital lower respiratory tract malformations. AB - In this article, the most important lower respiratory tract malformations are briefly reviewed, with special focus on those factors that may have some impact on the long-term respiratory outcome of specific lesions, like the amount of lung tissue resected, compensatory lung growth, lung hypoplasia, intensive care and mechanical ventilation. PMID- 16708309 TI - Communication training and antibiotic use in acute respiratory tract infections. A cluster randomised controlled trial in general practice. AB - A cluster-randomised controlled trial in general practice BACKGROUND: Physician patient communication plays a key role in treatment decisions in primary care. We aimed to reduce the antibiotic prescription rate for acute respiratory tract infections using a short training programme in patient-centred communication. METHODS: We conducted a cluster-randomised controlled trial in 45 general practices in Switzerland. Thirty physicians received evidence-based guidelines for the management of acute respiratory tract infections; 15 physicians randomised to the full intervention additionally received training in patient centred communication. A further 15 physicians, not randomised, served as a control to blind the physicians in the other two groups to the true comparison. The primary outcome was the antibiotic prescription rate reported by pharmacists. Secondary outcomes were patient satisfaction and enablement, re-consultation rates, days with restrictions, and days off work. 1108 adults with acute respiratory infections were screened between January and May 2004. Outcomes were measured in 837 consultations; 624 patients had follow-up interviews at 7 and 14 days. RESULTS: The antibiotic prescription rate reported by pharmacists was low in both full and limited intervention groups (13.5% and 15.7% respectively) but only half of the antibiotics were prescribed according to guidelines (53.8% and 53.1%). No significant differences were seen between the two randomised groups in primary and secondary outcomes. In both groups patient satisfaction was high (median score for both 68 out of 70). CONCLUSIONS: In this trial, patient-centred communication training did not reduce the rate of antibiotic prescriptions below an already unusually low level. Even with this low prescription rate, patient satisfaction with received care was high. PMID- 16708310 TI - Prevalence and influence of diagnostic tests for acute respiratory tract infections in primary care. AB - BACKGROUND: General practitioners (GPs) use diagnostic tests to help distinguish between viral and bacterial acute respiratory tract infections (ARTI). We investigated the prevalence of these tests, and how tests are associated with diagnosis, treatment and patient satisfaction. METHODS: As part of a clinical trial, 45 GPs screened 1108 patients with ARTI and collected information on signs and symptoms, diagnostic test results, and subsequent diagnosis and treatment. A sample of 636 patients was interviewed after 7 days and their opinions recorded. We used multivariate mixed models to estimate associations between the use of tests and (1) baseline characteristics, (2) subsequent antibiotic treatment, and (3) patient satisfaction. RESULTS: GPs carried out at least one test in 42% of the 1108 patients screened. The tests used were (percentage of patients): CRP (35%), leucocyte count (17%), rapid Strep A (9%), chest X-ray (5%), sinus X-ray (1%), and throat culture (1%). The use of tests was associated with increasing patient age, education, and degree of discomfort. Antibiotic therapy was strongly associated with a positive test, with odds ratios of 26 (95% CI, 10-67) for a CRP above 50 mg/l; 9.6 (95% CI, 3.6-26) for a leucocyte count above 10,000/microl; and 122 (4.4-3435) for a positive StrepA test. There was no evidence of an association between the use of tests and patient satisfaction. CONCLUSIONS: Nearly half of these patients with ARTI received a diagnostic test. Older patients, those with higher education and those in more discomfort were more likely to get tests. A positive test was strongly associated with antibiotic treatment. PMID- 16708311 TI - Increased prevalence of high Body Mass Index in patients presenting with pituitary tumours: severe obesity in patients with macroprolactinoma. AB - INTRODUCTION: Prolactinoma has been associated with obesity. As opposed to ACTH- and GH-secreting adenoma, the mechanism by which macroprolactinoma causes obesity has not been fully understood. Having seen patients with both prolactinoma and obesity and more recent literature on brain dopamine, dopamine 2 receptors and obesity, we re-evaluated the potential relationship between prolactinoma and obesity. METHODS: Data of patients with pituitary adenomas were collected retrospectively over a period of 20 years. 399 patients with well-documented pituitary adenomas and information about pre-treatment body mass index (BMI), age, sex, and tumour type were analysed. RESULTS: Elevated BMI (> or = 30 kg/m2) was observed in 8/36 patients (22.2%) with ACTH-producing tumours, in 15/70 (21.4%) with GH-producing tumours, in 25/100 (25%) with macroprolactinoma, in 8/81 (9.9%) with microprolactinoma, and in 18/105 (17.1%) with inactive macroadenomas. Macroprolactinoma patients had a mean BMI value (27.5 +/- 7.7 kg/m2) similar to that of patients with Cushing's disease (27.2 +/- 5.9 kg/m2) and acromegaly (27.4 +/- 4.4 kg/m2) and on average a significantly higher BMI value compared to that of patients with inactive macroadenomas (25.8 +/- 4.4 kg/m2) (95% CI 1.2, 4.4; p-value <0.001). Compared to the general population, the proportion of BMI > or = 30 kg/m2 in patients with macroprolactinoma was significantly higher (95% CI 0.1, 0.29; p-value <0.001). CONCLUSIONS: Average BMI in macroprolactinoma patients is significantly higher than BMI in patients with inactive adenomas. Macroprolactinoma is associated with increased frequency of obesity compared to the general population. We propose that in a subgroup of individuals obesity and macroprolactinoma may share a common basis, namely decreased dopamine 2 receptor-mediated actions. PMID- 16708312 TI - [Computer tomography of retroperitoneal fibrosis]. PMID- 16708322 TI - [Value of MR-guided localization and biopsy in breast lesions]. AB - Contrast-enhanced dynamic MR mammography can provide important additional diagnostic information when performed for certain indications. When suspicious lesions are identified on MR mammography and cannot be reproduced using other imaging modalities, a decision must be made as to its management, i. e. further diagnostic work-up. One possibility is the short-term follow-up of such findings, resulting in higher costs and a possible delay in the start of treatment of a malignant lesion. An alternative to a follow-up is an MR-guided intervention. MR compatible equipment has been developed for this purpose. This includes equipment specialized for percutaneous biopsy and preoperative localization. The following is an overview of the diagnostic value of MR-guided biopsy and preoperative localization including the relevant literature. PMID- 16708323 TI - [Cardiovascular whole-body MR imaging in patients with symptomatic peripheral arterial occlusive disease]. AB - PURPOSE: To examine patients with peripheral-arterial-occlusive-disease (PAOD) for systemic effects associated with atherosclerosis using a comprehensive state of-the-art whole-body MR examination protocol. The protocol comprises the assessment of the complete arterial vasculature (except coronary arteries), the brain, and the heart. MATERIALS AND METHODS: Multi-station whole-body 3D MR angiography was performed in sixty consecutive patients with clinical suspicion for PAOD at 1.5 T (Magnetom Avanto, Siemens, Erlangen, Germany). Functional and delayed enhancement cardiac images were acquired, as well as FLAIR images of the brain and TOF angiography of intracranial vessels. MR and DSA images were assessed by independent observers for atherosclerotic manifestations and other pathology. Sensitivity and specificity for the detection of vascular pathology was calculated for MR data using conventional DSA of the symptomatic region as standard-of-reference. RESULTS: Sensitivity and specificity for the detection of significant vascular stenosis (> 70 % luminal narrowing) was 94 % and 96 % (PPV 87 %, NPV 98 %). Significant microangiopathic tissue alterations (n = 7) and/or cerebral infarction (n = 18) were diagnosed in 23/60 patients. Thirty-eight of 60 patients presented with systolic left ventricular wall motion abnormalities. In 24 patients subendocardial or transmural delayed enhancement was detected in corresponding regions, indicating prior myocardial infarction. CONCLUSION: For patients with PAOD and suspected systemic atherosclerotic disease a comprehensive diagnosis of accompanying cardiovascular pathology and therefore staging of systemic atherosclerotic disease is feasible within one MR examination. PMID- 16708328 TI - [Airway management in infants]. AB - For a large number of anesthesiologists, airway management in infants is not part for their daily routine and thus represents a major challenge. In the present review the special anatomic and physiological features of the infant airways are summarized. Various technical aids for airway management in infants are described in the context of their development and utility in routine clinical practice. PMID- 16708329 TI - [The cardiac risk patient: introduction and epidemiology]. AB - Preexisting cardiovascular diseases are the most important risk factors in the perioperative period. They can cause complications that have a major impact on the morbidity and mortality of surgical patients. The current issue of AINS thus presents the actual state of knowledge about the pathophysiology, preoperative evaluation, and perioperative management of cardiac risk diseases in three practice-orientated review articles. Only sound knowledge of the pathophysiological background together with the diagnostic and therapeutic possibilities will allow the clinically active anesthesiologist to adapt the available therapeutic and preventative options in the best possible way to the individual patient and to avoid not only medical but also economically unnecessary and/or potentially harmful interventions. PMID- 16708330 TI - [Pathophysiology of coronary artery disease and heart failure]. AB - Comorbid cardiovascular diseases can significantly increase the risk for perioperative morbidity and mortality. Increasing age of surgical populations and increasing invasiveness of surgical procedures further contribute to a significantly increased perioperative cardiovascular complication rate. Treatment of cardiovascular comorbidities and prevention of cardiovascular complications can decrease the perioperative risk. For detection and treatment of cardiovascular risk patients a thorough understanding of the underlying pathophysiology is of utmost importance. This article therefore reviews clinically relevant features of cardiovascular diseases with an emphasis on coronary artery disease and heart failure. PMID- 16708331 TI - [Preoperative stratification of risks in patients with disorders of the cardiovascular system]. AB - Disorders of the cardiovascular system are among the most important accompanying diseases in operative medicine. These diseases increase the perioperative risk; the risk is assessed on the basis of the case history and tolerance of the patient as well as the extent and invasiveness of the surgical intervention. Depending on the results of this stratification further examination and therapy may be necessary. The aim of these mainly preventative measures is the highest possible preoperative hemodynamic stability. Besides pharmacological prevention, a therapy for hypertension or ischemia belongs among these measures. The selected anesthesia process must guarantee and efficient prevention of pain. PMID- 16708332 TI - [Preoperative identification of cardiac risk patients and perioperative management]. AB - The demographic trend challenges anaesthesiologists with a growing number of elderly requiring surgery. The anaesthetist needs to identify risk patients and to optimize his strategies for perioperative management. The present article gathers the current data and summarizes effective strategies for anaesthesia in patients with ischemic heart disease. PMID- 16708334 TI - Treatment of recurrent previously coiled anterior circulation aneurysm with minimally invasive keyhole craniotomy: report of two cases. AB - The use of minimally invasive techniques has not yet been reported for the treatment of recurrent aneurysms after coil embolization. A 47-year-old man with a long history of headaches had an anterior communicating aneurysm that had previously been coil embolized. Three-year follow-up angiography showed a significant recurrence. A 50-year-old woman with subarachnoid hemorrhage and acute visual loss underwent coil embolization of a large ophthalmic artery aneurysm, which recurred 3 months later. In both cases, a keyhole fronto-orbital one-piece craniotomy was performed. In the first patient, the aneurysm was clip ligated. The coil mass, which had eroded through the dome, was excised. In the second patient, the anterior clinoid was removed and the aneurysm was clip ligated. Postoperative angiography showed no residual aneurysm and no evidence of branch or parent vessel compromise in either patient. Both patients had an uncomplicated postoperative course. Recurrent previously coiled aneurysms are technically challenging to treat. A minimal fronto-orbital craniotomy provides a sufficiently capacious working space for successful treatment of some recurrent aneurysms of the anterior circulation. PMID- 16708333 TI - Endovascular treatment of small intracranial aneurysms: three alternatives to coil occlusion. AB - INTRODUCTION: Small intracranial aneurysms with a fundus diameter of 2 - 3 mm may rupture and are therefore potential targets for an endovascular approach in treatment. Currently available coil technology is less than optimal for the treatment of aneurysms within this size range. Even the smallest coils are sometimes too large. If such a minute coil can be introduced into a small aneurysm, the hemodynamic effect and the induced thrombosis are frequently inadequate to occlude the aneurysm sufficiently from the parent artery circulation. METHODS: Three technical alternatives for the endovascular treatment of small intracranial aneurysms not suitable for coil occlusion are illustrated with the following three case descriptions. RESULTS: Stent grafts are usable for the intracranial internal carotid artery and for the V4 segment. The stiffness of the stent and the high expansion pressures are the two major drawbacks. Coaxial deployment of two or more self-expanding porous stents can result in sufficient redirection of the blood flow to induce aneurysmal thrombosis. Deployment of multiple stents, however, may require several treatment sessions in order to allow for the integration of the stents into the vessel wall from session to session. A regular microcatheter can block aneurysmal inflow in aneurysms with a very narrow neck. This allows the occlusion of the aneurysm with an appropriate amount of highly concentrated, rapidly polymerizing glue. Polymer emboli may result from excessive or rapid glue injection. CONCLUSION: The available coil technology has inherent limitations in the treatment of very small intracranial aneurysms. Liquid embolic agents and stent-based extrasaccular treatment strategies may provide solutions for these challenging lesions. PMID- 16708335 TI - Percutaneous endoscopic transforaminal lumbar discectomy: a critical appraisal. AB - Percutaneous endoscopic transforaminal lumbar discectomy (PETD) is one of the surgical techniques for the treatment of the lumbar disc herniations. Fenestration of the annulus and partial resection of the nuclear substance are achieved percutaneously by a posterolateral approach under local anaesthesia. The results of the first 42 patients are evaluated regarding the learning curve and indications for this procedure. The mean follow-up time was 15 months. Excellent and good results were evaluated as successful and the overall success rate is 77 %. All six patients with foraminal disc herniations in whom a free fragment could be removed had excellent results. Military personnel can return to work quickly without deficits with the use of this technique. PMID- 16708336 TI - A New full-endoscopic technique for the interlaminar operation of lumbar disc herniations using 6-mm endoscopes: prospective 2-year results of 331 patients. AB - Even with good results, conventional disc operations may result in consecutive damage due to traumatisation. Endoscopic techniques have become the standard in many areas because of the advantages they offer in surgical technique and in rehabilitation. The transforaminal operation is the most common full-endoscopic procedure in surgery of the lumbar spine. It is frequently necessary to reach the spinal canal directly in order to achieve sufficient resection of lumbar disc herniations. Even in using a lateral approach, the authors recognise the clear limitations of the transforaminal procedure. The objective of this prospective study was to examine the technical possibility of a full-endoscopic interlaminar access. The focus was on questions of sufficient decompression, as well as advantages and disadvantages of the minimally invasive procedure. 331 patients were followed for 2 years. The results show that 82 % reported no longer having leg pain, and 13 % had only occasional pain. The decompression results are equivalent to those of conventional procedures. Traumatisation of both the access pathway and the spinal canal structures was reduced. Epidural scarring was minimised. The recurrence rate was 2.4 %. No serious surgical complications were observed. The authors view the technique described, which offers the advantage of a truly minimally invasive procedure, as a sufficient and safe alternative to conventional procedures, when the appropriate indication criteria are heeded. There are technical problems because of the small instruments. In conjunction with the transforaminal procedure, this is an expansion of the spectrum for full endoscopic surgery of lumbar disc herniations. PMID- 16708337 TI - Endoscopic endonasal approach for cerebrospinal fluid fistulae. AB - Different techniques have been proposed to repair cerebrospinal fluid rhinorrhea. Advances in nasal surgery led to a high success rate and low morbidity for the endonasal approach. It has become the favorite route for treating cerebrospinal fluid leaks of the anterior skull base. Better results have been obtained with the improvement of rigid endoscopes and intrathecal sodium fluorescein. In a prospective study, twenty-four patients with cerebrospinal fluid rhinorrhea were evaluated and treated by endoscopic endonasal surgery. In all cases intrathecal sodium fluorescein enabled a precise localization of the bone defect. The most common causes of CSF rhinorrhea were traumatic (8 cases, 33 %), spontaneous (6 cases, 25 %), and iatrogenic (5 cases, 20.8 %). Preoperative radiological evaluations (plane CT, CT cisternogram and MRI) showed the exact site and size of the defect in all patients. The most common site of leakage was the ethmoidal roof-cribriform plate. Primary closure was achieved in all patients. There were no major operative or postoperative complications. The endoscopic endonasal approach can be considered the first choice in the treatment of cerebrospinal fluid rhinorrhea. PMID- 16708338 TI - Image-guided endonasal transsphenoidal microsurgical treatment of recurrent microadenomas of the pituitary gland. AB - BACKGROUND: Neuronavigation is a commonly used technology that provides continuous, three-dimensional information for the precise localization of and surgical trajectory to brain lesions. This study was performed to evaluate the role that navigation can play in assisting microsurgical transsphenoidal surgery for precise localization and removal of recurrent pituitary tumours while simultaneously preserving pituitary gland function. METHOD: During a 6-month period -- July 2004 until December 2004 -- 9 patients with recurrent pituitary tumours (5 female and 4-male) were treated with navigation-guided transsphenoidal microsurgical resection. Surgery was performed via a paraseptal or endonasal transsphenoidal approach. The navigation system Vector Vision (Brain Lab, Heimstetten, Germany) allowed precise localization of the tumours (7 hormonal active and 2 inactive microadenomas) in respect to the pituitary gland, the carotid arteries and the cavernous sinus. RESULTS: Postoperative MRI investigations of the 9 patients treated with image-guided transsphenoidal microsurgery, showed total tumour removal in 7 (77 %) patients and subtotal removal in 2 patients (23 %). One patient (11 %) developed a cerebral spinal fluid (CSF) leak and was treated conservatively. One patient (11 %) had preoperative insufficiency of the corticotrope axis which remained unchanged postoperatively. Of the remaining 8 patients who did not have preoperative endocrinological disturbance, only one (12 %) developed postoperative insufficiency of the corticotrope axis. Out of the 7 patients with hormone active tumours, 5 (72 %) patients showed no more postoperative hormonal activity. CONCLUSION: Microneurosurgical transsphenoidal techniques combined with image guided systems can precisely define the localization and removal of lesions in the sella region with respect to the margins of important anatomical structures in the neighbourhood and the endocrinological functionality of the pituitary gland. PMID- 16708339 TI - Analysis of surgical management of calvarial tumours and first results of a newly designed robotic trepanation system. AB - This study was performed to evaluate the surgical strategy in patients with calvarial tumours, in order to design and modify a robot-assisted trepanation system. A total of 75 patients underwent craniectomy for the treatment of calvarial tumours during the 10-year period from 1993 to 2002. The patients' complaints, the size, location and histology of the tumour, and the various cranioplasty techniques used were analysed retrospectively. In a second procedure several craniectomies at typical locations according to the study's results were performed in a laboratory setting using a hexapod robotic tool, constructed at the Helmholtz-Institute, RWTH Aachen University, and plastic model heads. The workflow was documented and the reproducibility and the accuracy of the procedure were registered. A total of 83 surgical procedures were performed on 75 patients. The majority (87 %) of lesions treated surgically were located in the frontal, temporal and anterior parts of the parietal region. Histological examination revealed benign lesions in 66 % of the patients and dural involvement in 46 %. According to these results craniectomies were performed using the robotic system. Mean positioning accuracy of the robotic system while milling was 0.24 mm, with a standard deviation of 0.04 mm, and maximum error under 1 mm. Craniectomies leaving a 1-mm layer of the tabula interna intact to ensure a healthy dura were performed in several regions successfully. The majority of calvarial tumours, requiring surgical treatment in our patients, were located in cosmetically relevant areas in which drilling can be carried out with the robotic trepanation system. Consequently, the surgical approach had to be planned carefully in order to achieve a good cosmetic outcome. PMID- 16708340 TI - Medial microsurgical approach to the orbit: an anatomic study. AB - The authors have identified and described three medial approaches to the intraorbital portion of the optic nerve in cadavers. The structures exposed via the superior, inferior or central approaches were approached through the medial orbital wall. The superior approach in which the medial rectus muscle is retracted inferiorly exposes the superomedial aspect of the orbit. The inferior approach in which the medial rectus muscle is retracted superiorly, provided the inferomedial aspect of the optic nerve from the globe to the muscle cone. The central approach in which the ethmoidectomy was performed and the medial rectus muscle was disinserted and retracted, provided excellent exposure of the structures of the medial part of the orbit and the optic nerve. There were no structures blocking access to the two third anterior optic nerve and medial surface of the globe. The structures encountered in these approaches are demonstrated under magnification in cadavers, and the exposure of the optic nerve is discussed. PMID- 16708341 TI - CyberKnife stereotactic radiotherapy for patients with malignant glioma. AB - OBJECTIVE: The CyberKnife is a new frameless image-guided radiosurgical modality. The authors report on their experience using the CyberKnife in 25 patients with malignant gliomas. METHODS: Twenty-five patients with histologically proven malignant gliomas (18 glioblastoma: GB, 7 anaplastic astrocytoma: AA) were treated with the CyberKnife at Konan St. Hill Hospital between June 1998 and November 2002. CyberKnife therapy was performed on 44 lesions (31 GB lesions, 13 AA lesions) in the 25 patients. The median target volume was 19.1 mL (range: 0.3 90.2). The median prescribed dose was 20.3 Gy (range: 13.9 - 26.4). Patient-, tumor-, and treatment-related variables were analyzed by univariate analysis, and survival curves were generated by the Kaplan-Meier product limit. RESULTS: In the 18 GB patients, the median survival after diagnosis was 20.7 months (82.6 weeks) with a mean follow-up of 85.7 weeks. Of the 7 AA patients, 6 were alive at the time of analysis with follow-up periods ranging from 11.4 to 52.8 months. Patients younger than 70 years had a median survival after diagnosis of 37.1 months, compared to 12.4 months for older patients (p = 0.003). Similarly, patients with well-controlled lesions had a median survival after diagnosis of 39.8 months compared to 16.0 months for those with uncontrolled lesions (p = 0.031). Late delayed radiation necrosis was seen in 1 GB patient. No other patient suffered acute or delayed neurological morbidity after CyberKnife therapy. CONCLUSION: This is the first report of CyberKnife stereotactic radiotherapy applied to the treatment of malignant gliomas. The frameless and painless CyberKnife stereotactic radiotherapy has the potential to be as useful for treatment of malignant glioma as other radiosurgical modalities. PMID- 16708342 TI - Management of intracranial germ cell tumors presenting with rapid deterioration of consciousness. AB - OBJECTIVES: The value of surgical intervention in the management of intracranial germ cell tumors remains controversial. PATIENTS: We report three patients who presented with rapid deterioration of consciousness but resulted in complete remission of the tumor after emergency surgery for both diagnostic (biopsy) and therapeutic (for hydrocephalus) purposes followed by prompt initiation of radiochemotherapy. CONCLUSION: For management of these rapidly deteriorating patients, immediate histological verification and avoidance of delay in the induction of radiochemotherapy are essential. Thus, emergency surgery with a less invasive procedure is the first choice of treatment, i. e., endoscopic surgery for pineal region tumor and CT-guided biopsy for basal ganglia tumor. PMID- 16708343 TI - Modified surgical technique for the treatment of idiopathic spinal cord herniation. AB - OBJECTIVE AND IMPORTANCE: Spinal cord herniation is not a common disease and only 80 cases have been reported to the best of our knowledge. The treatment of this lesion is thought to be difficult and a better surgical technique has not been established. CLINICAL PRESENTATION: A 57-year-old man presented with gradually worsening spastic gait and hypoesthesia in the bilateral lower extremities suspected to be due to Brown-Sequard syndrome. Magnetic resonance imaging showed a thoracic spinal cord herniation into the anterior hiatus at T2 - 3. INTERVENTION: A modified technique of closure of the hiatus in front of the herniation was performed. A surgical artificial dural membrane was introduced between the herniated portion of the thoracic cord and the dural hiatus and was placed to cover the whole dural defect in order to minimize the operative procedure at the ventral side of the spinal cord. On subsequent magnetic resonance imaging, the herniation was remarkably improved and patient's symptoms were abolished. CONCLUSION: The direct widening of the dural defect has been reported to be the only treatment providing a good outcome. We suggest that our modified technique for its closure can be as safe and effective as the direct widening method. PMID- 16708344 TI - Endoscopic aqueductoplasty with and without stent. PMID- 16708345 TI - Event history analysis and the cross-section. AB - Examples are given of problems in event history analysis, where several time origins (generating calendar time, age, disease duration, time on study, etc.) are considered simultaneously. The focus is on complex sampling patterns generated around a cross-section. A basic tool is the Lexis diagram. PMID- 16708346 TI - Analysing capture--recapture data when some variables of heterogeneous catchability are not collected or asked in all registrations. AB - Registrations in epidemiological studies suffer from incompleteness, thus a general consensus is to use capture-recapture models. Lately, there has been a thrust to incorporate covariates which relate to the capture probabilities in order to improve the estimate of population size. Current approaches utilize covariates observed in all the pertinent registrations. In epidemiological applications, one often encounters capture-recapture data where some covariates are measured by some but not by all of the registrations under scrutiny. In this article, the usefulness of these types of covariates is evaluated and furthermore a methodology for analysing such capture-recapture data is presented. The methodology is illustrated by applying it to data on neural tube defects in the Netherlands. PMID- 16708347 TI - Variable selection and raking in propensity scoring. AB - This paper discusses some practical issues in applying propensity scoring in the context of endpoint analysis in a pre-/posttest longitudinal design with an ordinal measure of treatment intensity and a high-dimensional potential covariate space: how many covariates to include in propensity models; how to evaluate the adequacy of tentative propensity models; and how to tailor models to provide hypercontrol on a limited subset of covariates. These issues arose in the evaluation of a health communication program. PMID- 16708348 TI - Inferences on standardized mean difference: the generalized variable approach. AB - The standardized mean difference has been widely used as the most common index of effect magnitude in many applied fields. In this paper, we propose a novel approach using the concept of generalized variable for the confidence interval estimation and hypothesis testing of standardized mean difference. Furthermore, we extend this approach to compare standardized mean differences between two studies or between two strata. Simulation results demonstrate that the proposed approach can provide confidence intervals with excellent coverage properties and can perform hypothesis testing with satisfactory type-I error control. PMID- 16708349 TI - A comparison of the ability of different propensity score models to balance measured variables between treated and untreated subjects: a Monte Carlo study. AB - The propensity score--the probability of exposure to a specific treatment conditional on observed variables--is increasingly being used in observational studies. Creating strata in which subjects are matched on the propensity score allows one to balance measured variables between treated and untreated subjects. There is an ongoing controversy in the literature as to which variables to include in the propensity score model. Some advocate including those variables that predict treatment assignment, while others suggest including all variables potentially related to the outcome, and still others advocate including only variables that are associated with both treatment and outcome. We provide a case study of the association between drug exposure and mortality to show that including a variable that is related to treatment, but not outcome, does not improve balance and reduces the number of matched pairs available for analysis. In order to investigate this issue more comprehensively, we conducted a series of Monte Carlo simulations of the performance of propensity score models that contained variables related to treatment allocation, or variables that were confounders for the treatment-outcome pair, or variables related to outcome or all variables related to either outcome or treatment or neither. We compared the use of these different propensity scores models in matching and stratification in terms of the extent to which they balanced variables. We demonstrated that all propensity scores models balanced measured confounders between treated and untreated subjects in a propensity-score matched sample. However, including only the true confounders or the variables predictive of the outcome in the propensity score model resulted in a substantially larger number of matched pairs than did using the treatment-allocation model. Stratifying on the quintiles of any propensity score model resulted in residual imbalance between treated and untreated subjects in the upper and lower quintiles. Greater balance between treated and untreated subjects was obtained after matching on the propensity score than after stratifying on the quintiles of the propensity score. When a confounding variable was omitted from any of the propensity score models, then matching or stratifying on the propensity score resulted in residual imbalance in prognostically important variables between treated and untreated subjects. We considered four propensity score models for estimating treatment effects: the model that included only true confounders; the model that included all variables associated with the outcome; the model that included all measured variables; and the model that included all variables associated with treatment selection. Reduction in bias when estimating a null treatment effect was equivalent for all four propensity score models when propensity score matching was used. Reduction in bias was marginally greater for the first two propensity score models than for the last two propensity score models when stratification on the quintiles of the propensity score model was employed. Furthermore, omitting a confounding variable from the propensity score model resulted in biased estimation of the treatment effect. Finally, the mean squared error for estimating a null treatment effect was lower when either of the first two propensity scores was used compared to when either of the last two propensity score models was used. PMID- 16708350 TI - Testing the equality of two survival functions with right truncated data. AB - To compare the survival functions based on right-truncated data, Lagakos et al. proposed a weighted logrank test based on a reverse time scale. This is in contrast to Bilker and Wang, who suggested a semi-parametric version of the Mann Whitney test by assuming that the distribution of truncation times is known or can be estimated parametrically. The approach of Lagakos et al. is simple and elegant, but the weight function in their method depends on the underlying cumulative hazard functions even under proportional hazards models. On the other hand, a semi-parametric test may have better efficiency, but it may be sensitive to misspecification of the distribution of truncation times. Therefore, this paper proposes a non-parametric test statistic based on the integrated weighted difference between two estimated survival functions in forward time. The comparative results from a simulation study are presented and the implementation of these methods to a real data set is demonstrated. PMID- 16708351 TI - Mutational dynamics in human tumors confirm the neutral intrinsic instability of the mitochondrial D-loop poly-cytidine repeat. AB - Somatic mutations at a mitochondrial noncoding polycytidine (C)(n) repeat (polyC) have been associated with tumor progression. We analyzed whether these alterations are due to the inherent mutability of repeated sequences. Insertion and deletion mutations were found in colon (n = 114), stomach (n = 105), endometrium (n = 53), breast (n = 45), lung (n = 35), and prostate (n = 20) tumors. The mutation frequency in colon, gastric, and endometrial tumors was 23, 17, and 11%, respectively, which paralleled the relative extent of microsatellite instability in long mononucleotide repeats observed in tumors with mismatch repair deficiency (colon > stomach > endometrium, relative ratio 10:8:4). Colon tumors with mutations of more than one nucleotide were more advanced in tumor progression. Further, two tumors showing a T > C mutation that restored the homopolymeric repeat, harbored sequential deletion mutations of up to 4 and 6 nucleotides. These results illustrate that the increased mutability of repeated mitochondrial sequences is dependent on the repetitive structure of the DNA molecule and suggest that mutations in the (C)(n) repeat, whether homoplasmic or not, and by extrapolation, mitochondrial mutations in general, are not the result of selective pressure during tumorigenesis. We also suggest that the (C)(n) repeat may be used as an universal molecular clock to estimate the relative mitotic history of tumors. PMID- 16708352 TI - CpG island methylation of genes accumulates during the adenoma progression step of the multistep pathogenesis of colorectal cancer. AB - Genetic alterations occur during the adenoma-carcinoma sequence of colon cancer formation and drive the initiation and progression of colon cancer formation. The aberrant methylation of genes is an alternate, epigenetic mechanism for silencing tumor suppressor genes in colon cancer. The aim of this study was to determine on a global and gene-specific level the role of CpG island methylation in the initiation and progression of colon cancer. Consequently, we assessed the frequency of gene methylation in tumors representative of the commonly recognized histological steps of the adenoma-carcinoma progression sequence through the analysis of eight genes previously identified to be methylated in colon cancer, MGMT, HLTF, MLH1, p14(ARF), CDKN2A, TIMP3, THBS1, and CDH1. We observed that the proportion of tumors carrying methylated alleles increased from adenomas to adenocarcinomas but that the proportion of tumors with methylated alleles was not different between adenocarcinomas and metastases (69% versus 90%, P = 0.01 and 90% versus 81%, P > 0.05). The most substantial difference occurred between early and advanced adenomas (47% versus 84%, P = 0.018). Furthermore, we observed that the frequency of gene methylation at the different steps of the progression sequence varied between genes. Thus, the aberrant methylation of genes appears to increase most significantly during the progression of early adenomas to advanced adenomas, and the frequency of specific gene methylation at the different steps of the adenoma-carcinoma progression sequence varies in a gene-specific fashion. PMID- 16708353 TI - RNA interference-based functional dissection of the 17q12 amplicon in breast cancer reveals contribution of coamplified genes. AB - DNA amplification is a frequent occurrence in cancer genomes. While tumor amplicons may harbor known oncogenes "driving" amplification, amplicons rarely comprise only single genes. The potential functional contribution of coamplified genes remains largely unexplored. In breast cancer, 20-30% of tumors exhibit amplification within chromosome band 17q12, containing the ERBB2 oncogene. Analysis of array-based comparative genomic hybridization and expression profiling data indicate that the minimum region of recurrent amplification (i.e., the amplicon "core") at 17q12 includes two other genes, GRB7 and STARD3, which exhibit elevated expression when amplified. Western blot analysis confirms overexpression of each at the protein level in breast cancer cell lines SKBR3 and BT474 harboring amplification. In these cell lines (but not in control MCF7 breast cancer cells lacking 17q12 amplification), targeted knockdown of ERBB2 expression using RNA interference (RNAi) methods results in decreased cell proliferation, decreased cell-cycle progression, and increased apoptosis. Notably, targeted knockdown of either GRB7 or STARD3 also leads to decreased cell proliferation and cell-cycle progression, albeit to a lesser extent compared with ERBB2 knockdown. We conclude that the amplification and resultant overexpression of genes coamplified with ERBB2 at 17q12 can contribute to proliferation levels of breast cancer cells. Our findings validate the utility of RNAi in the functional interrogation of tumor amplicons, and provide evidence for a contribution of coamplified genes to tumor phenotypes. PMID- 16708354 TI - The risk of secondary malignancies over 30 years after the treatment of non Hodgkin lymphoma. AB - BACKGROUND: Survivors of non-Hodgkin lymphoma (NHL) are at increased risk for developing secondary malignancies. For the current study, the authors quantitated this risk in a group of NHL survivors over 30 years of follow-up. METHODS: Standardized incidence ratios (observed-to-expected [O/E] ratio) and absolute excess risk of secondary malignancies were assessed in 77,876 patients who were diagnosed with NHL between 1973 and 2001 from centers that participated in the National Cancer Institute's Surveillance, Epidemiology, and End Results Program. RESULTS: There were 5638 patients who developed secondary malignancies, significantly more than the endemic rate (O/E, 1.14; P < .001). Overall, irradiated patients had a similar risk of secondary malignancies compared with unirradiated patients (relative risk, 1.04; 95% confidence interval, 0.98-1.10; P = .21). Irradiated patients had excess risk for sarcomas, breast cancers, and mesothelioma compared with unirradiated survivors (P < .05). Patients age <25 years at the time of their NHL diagnosis had the highest relative increased risk (no radiation: O/E, 2.1; P < .05; radiation: O/E, 4.51; P < .05). Overall, no statistical difference was observed for secondary cancer incidence between females and males (O/E, 1.12 vs. 1.15, respectively). Female survivors of NHL were less likely to develop breast cancer than the general population (O/E, 0.85; P < .05), but women age <25 years at the time of their NHL diagnosis were more likely to develop breast cancer (no radiation: O/E, 2.1; P < .05; radiation: O/E, 4.51; P < .05). CONCLUSIONS: The overall risk of secondary malignancies was increased for NHL survivors and varied according to age at NHL diagnosis, gender, and treatment. PMID- 16708355 TI - Mononuclear cell infiltration in clear-cell renal cell carcinoma independently predicts patient survival. AB - BACKGROUND: The impact of mononuclear cell infiltration on renal cell carcinoma (RCC) biology has been controversial, previously reported to be associated with either a favorable or unfavorable prognosis. The objective of the current study was to evaluate associations between mononuclear cell infiltration in routinely prepared paraffin-embedded specimens with survival in patients with clear-cell RCC. METHODS: A total of 306 patients were identified treated with nephrectomy for clear-cell RCC between 1990 and 1994. A single urologic pathologist, blinded to patient outcome, reviewed the specimens and quantified the extent of mononuclear cell infiltration as absent, focal, moderate, or marked. Cancer specific survival was estimated using the Kaplan-Meier method. Associations of mononuclear cell infiltration with death from RCC were assessed using Cox proportional hazards regression models. RESULTS: At last follow-up, 173 of the 306 patients studied had died, including 96 patients who died from RCC. Mononuclear cell infiltration was absent in 165 (54%), focal in 70 (23%), moderate in 53 (17%), and marked in 18 (6%). Univariately, patients with specimens that had mononuclear cell infiltration were over 2 times more likely to die from RCC compared with patients whose specimens exhibited no mononuclear cell infiltration (risk ratio, 2.63; P < .001). After adjusting for the Mayo Clinic SSIGN (stage, size, grade, and necrosis) score, patients with specimens that had mononuclear cell infiltration exhibited a significantly increased likelihood of dying from RCC compared with patients whose specimens had no mononuclear cell infiltration (risk ratio, 1.61; P = .028). CONCLUSIONS: Mononuclear cell infiltration is associated with death from RCC even after multivariate adjustment. Routine documentation of mononuclear cell infiltration is recommended during the pathologic assessment of RCC. PMID- 16708356 TI - Impact of fellowship training on pathologic outcomes and complication rates of radical prostatectomy. AB - BACKGROUND: The study was conducted to assess the results of radical prostatectomy (RP) performed by fellowship-trained surgeons in the first year of independent practice. METHODS: A prospective cohort study of 66 men who underwent RP performed by 2 recently graduated fellowship-trained surgeons (C.J.R., n = 27; A.M.K., n = 39) in their first year of independent practice was undertaken. Preoperative, operative, and postoperative data were collected and analyzed. Median follow-up of the cohort is 12.5 months. RESULTS: The median patient age was 61.2 +/- 6.9 years (range, 44-74 years), the median prostate-specific antigen level was 5 ng/mL (range, 1.2-39.4 ng/mL), and the median prostate biopsy determined Gleason score was 7. Of the 66 men, 25 (38%) underwent a bilateral nerve-sparing RP, 20 (30%) underwent a unilateral nerve-sparing RP, and 21 (32%) underwent a nonnerve-sparing procedure. Forty-two men (63%) underwent a pelvic lymph node dissection. The median operative time was 201 minutes. Median estimated blood loss was 734 mL (range, 300-1600 mL). There were 4 major complications--a pulmonary embolism in 3 patients and an intraoperative rectal injury in 1. Pathologic classification was as follows: pT2, 74%; pT3a, 23%; pT3b, 2%; and pN+, 2%. The positive margin rate was 14% overall and only 2% in men with pT2 disease. CONCLUSIONS: Results of RP performed by fellowship-trained surgeons in their first year of practice compare favorably with results of RP in a large series reported by more experienced surgeons. Being trained in an environment where an experienced surgeon serves as first assistant to the trainee appears to abbreviate the learning curve associated with this procedure. PMID- 16708357 TI - Epsilon aminocaproic acid reduces transfusion requirements in patients with thrombocytopenic hemorrhage. AB - BACKGROUND: Epsilon aminocaproic acid (EACA) is an antifibrinolytic drug that has been used to control hemorrhage by stabilizing the thrombus. It has been used in thrombocytopenic patients largely on an empiric basis. METHODS: Concerns regarding side effects have limited the use of this drug. The authors reviewed their experience with EACA at the Cleveland Clinic Foundation from 1997 to 2003. RESULTS: Of 77 patients with thrombocytopenic hemorrhage, 51 (66%) patients achieved a complete response and 13 (17%) patients achieved a partial response, resulting in a decrease in platelet and red blood cell transfusions. Adverse effects were manageable in this set of patients with severe underlying disease. CONCLUSIONS: Based on this experience, EACA may be a valuable adjunctive therapy in the treatment of patients with thrombocytopenic hemorrhage. PMID- 16708358 TI - The model for end-stage liver disease-based Japan Integrated Scoring system may have a better predictive ability for patients with hepatocellular carcinoma undergoing locoregional therapy. AB - BACKGROUND: The Japan Integrated Scoring (JIS) system was revealed as a better model for outcome prediction compared with the Cancer of Liver Italian Program system for hepatocellular carcinoma (HCC), and the Model for End-Stage Liver Disease (MELD) was better as a prognostic predictor for patients with cirrhosis compared with the Child-Turcotte-Pugh (CTP) system, which is a parameter used in the JIS system. The objective of the current study was to investigate the performance of the modified MELD-based JIS system. METHODS: In the modified JIS system, the CTP class in the original JIS was replaced with MELD cut-off scores of <10, 10 to 14, and >14. The modified JIS system was compared with the original system in 276 patients with HCC who underwent locoregional therapy (transarterial chemoembolization or percutaneous injection). RESULTS: The mean +/- standard error original JIS score was 1.8 +/- 1.0 (range, 0-4), compared with 2.0 +/- 1.1 (range, 0-5) for the modified JIS system (P < .001). Using mortality as the endpoint, the area under receiver operating characteristic curve (AUC) for the modified JIS system was 0.804 compared with 0.741 for the original JIS system (P = .008) at 12 months, and the AUC was 0.853 and 0.765, respectively (P < .001), at 24 months. Survival analysis showed that the modified JIS system had a better discriminatory ability for patients in different score groups and was more accurate for outcome prediction in the Cox multivariate model. CONCLUSIONS: The current results indicated that the MELD-based, modified JIS system has improved predictive ability compared with the original system and is a more feasible model for clinical staging in patients with HCC who are undergoing locoregional therapy. PMID- 16708359 TI - Brief cognitive-behavioral audiotape interventions for cancer-related pain: Immediate but not long-term effectiveness. AB - BACKGROUND: Few studies have evaluated cognitive-behavioral interventions as an adjunct treatment for chronic cancer-related pain. A randomized clinical trial was performed evaluating the efficacy of 3 brief cognitive-behavioral techniques: relaxation, distraction, and positive mood interventions. METHODS: Fifty-seven patients with chronic cancer-related pain taking opioid medications were randomly assigned to either the relaxation, distraction, positive mood, or waiting-list control group. The patients in the 3 intervention groups received audiotapes of the cognitive-behavioral technique and were asked to practice regularly at home. The tapes were supplemented with written instructions and follow-up telephone calls. RESULTS: Patients in the relaxation and distraction groups reported significantly reduced pain intensity immediately after listening to the tapes. The pain reduction was not maintained, however. At the 2-week follow-up assessment, no significant differences in pain intensity or interference were found among the treatment groups. The groups also did not differ with regard to secondary outcome measures assessing quality of life, mood, self-efficacy, and other symptoms. The results of qualitative interviews indicated that patients often had difficulty focusing on the audiotapes and preferred their own methods of pain reduction. CONCLUSIONS: Brief relaxation and distraction audiotape interventions produced immediate pain reductions but not longer-term pain relief. Additional research with a more individualized intervention is needed to evaluate cognitive-behavioral interventions for cancer pain control. PMID- 16708360 TI - Surface-enhanced laser desorption/ionization time-of-flight mass spectrometry serum protein profiling to identify nasopharyngeal carcinoma. AB - BACKGROUND: Diagnosis of nasopharyngeal carcinoma (NPC) at an early disease stage is important for successful treatment and improving the outcome of patients. The use of serum protein profiles and a classification tree algorithm were explored to distinguish NPC from noncancer. METHODS: Serum samples were applied to metal affinity protein chips to generate mass spectra by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS). Protein peak identification and clustering were performed using the Biomarker Wizard software. Proteomic spectra of serum samples from 50 NPC patients and 54 noncancer controls were used as a training set and a classification tree with 6 distinct protein masses was generated by using Biomarker Pattern software. The validity of the classification tree was then challenged with a blind test set including another 20 NPC patients and 25 noncancer controls. RESULTS: The software identified an average of 93 mass peaks/spectrum and 6 of the identified peaks were used to construct the classification tree. The classification tree correctly determined 83% (123 of 149) of the test samples with 83% (58 of 70) of the NPC samples and 82% (65 of 79) of the noncancer samples. In a combination of the serum protein profiles with Epstein-Barr (EBV) nuclear antigen 1 (EBNA1 IgA) test, the diagnostic sensitivity and specificity were increased to 99% and 96%, respectively. CONCLUSIONS: The results suggest that SELDI-TOF-MS serum protein profiles could discriminate NPC from noncancer. The combination of serum protein profiles with an EBV antibody serology test could further improve the accuracy of NPC screening. PMID- 16708361 TI - Iloprost for the attenuation of ischaemia/reperfusion injury in a distant organ. AB - The objective of this study was to investigate antioxidant and cytoprotective properties of iloprost in a distant organ after ischaemia reperfusion injury. Male Wistar rats were divided into two groups. After application of anesthaesia both hindlimbs were occluded. A 2-h reperfusion procedure was carried out after 60 min of ischemia. Study group (STU) rats (n=10) received 10 microg kg(-1) iloprost in 1 ml of saline from the tail vein 10 min before reperfusion. Control (CON) group rats (n=10) received an equal amount of saline. The rats were sacrificed by injection of a high dose of thiopentone sodium. Blood and tissue samples (right kidneys) were taken for analysis. Differences in malondialdehyde (MDA), myeloperoxidase (MPO), Na+-K+ ATPase and total antioxidant capacity (TAC) between the groups were analysed. MPO, MDA and TAC levels in the sera of CON and STU groups were 1.60+/-0.26 U l(-1), 11.42+/-5.23 nmol ml(-1), 8.30 x 10(-2)+/- 3.93 x 10(-2) nmol ml(-1) h(-1) and 1.07+/-0.11 U l(-1), 7.60+/-1.81 nmol ml(-1) and 0.15+/-3.23 x 10(-2) nmol ml(-1) h(-1) (p=0.0001, p=0.043 and p=0.0001 respectively). MPO, ATPase and MDA levels in kidneys for CON and STU groups were 1.24+/-0.58 U g(-1), 85.70+/-52.05 nmol mg(-1), 17.90+/-7.40 nmol ml(-1) and 0.78+/-0.31 U g(-1), 195.90+/-56.13 nmol mg(-1) and 10.10+/-0.99 nmol ml(-1) (p=0.046, p=0.0001 and p=0.009 respectively). When given prior to reperfusion, the positive effect of iloprost in the attenuation of distant organ reperfusion injury has been demonstrated. PMID- 16708362 TI - Tolerance for random recombination of domains in prokaryotic and eukaryotic translation systems: Limited interdomain misfolding in a eukaryotic translation system. AB - It has been proposed that eukaryotic translation systems have a greater capacity for cotranslational folding of domains than prokaryotic translation systems, which reduces interdomain misfolding in multidomain proteins and, therefore, leads to tolerance for random recombination of domains. However, there has been a controversy as to whether prokaryotic and eukaryotic translation systems differ in the capacity for cotranslational domain folding. Here, to examine whether these systems differ in the tolerance for the random domain recombination, we systematically combined six proteins, out of which four are soluble and two are insoluble when produced in an Escherichia coli and a wheat germ cell-free protein synthesis systems, to construct a fusion protein library. Forty out of 60 two domain proteins and 114 out of 120 three-domain proteins were more soluble when produced in the wheat system than in the E. coli system. Statistical analyses of the solubilities and the activities indicated that, in the wheat system but not in the E. coli system, the two soluble domains comprised mainly of beta-sheets tend to avoid interdomain misfolding and to fold properly even at the neighbor of the misfolded domains. These results demonstrate that a eukaryotic system permits the concomitance of a wider variety of domains within a single polypeptide chain than a prokaryotic system, which is probably due to the difference in the capacity for cotranslational folding. This difference is likely to be related to the postulated difference in the tolerance for random recombination of domains. PMID- 16708363 TI - Molecular dynamics simulation of Leishmania major surface metalloprotease GP63 (leishmanolysin). AB - One of the molecular factors contributing to Leishmania sp. virulence and pathogenesis is the major surface metalloprotease GP63, alternatively called leishmanolysin, MSP, and PSP (EC 3.4.24.36). Here, the molecular dynamics simulation of Leishmania major GP63 in water at pH 7 is reported. Upon solvation, GP63 undergoes a sharp structural relaxation with respect to the crystal structure. The fluctuation pattern occurs essentially in solvent-exposed nonstructured regions. By contrast, the active site turns out to be rigid. Essential dynamics and dynamic-domain analyses, both carried out on the equilibrated portion of GP63, show that the fingerprint fluctuations of GP63 are practically characterized by the motion of a large part of the N-terminal domain. These results appear to be in line with substrate recognition and (pro)enzyme activation played by the N-terminal domain of GP63. A systematic analysis among a series of 10 homologs of GP63 also shows that the residues involved in the interdomain bending result highly conserved. This finding also suggests possible relationship between the maintainance of proteolytic activity and the similarity of the dynamical properties of the related enzymes. PMID- 16708364 TI - Minimizing false positives in kinase virtual screens. AB - In spite of recent improvements in docking and scoring methods, high false positive rates remain a common issue in structure-based virtual screening. In this study, the distinctive features of false positives in kinase virtual screens were investigated. A series of retrospective virtual screens on kinase targets was performed on specifically designed test sets, each combining true ligands and experimentally confirmed inactive compounds. A systematic analysis of the docking poses generated for the top-ranking compounds highlighted key aspects differentiating true hits from false positives. The most recurring feature in the poses of false positives was the absence of certain key interactions known to be required for kinase binding. A systematic analysis of 444 crystal structures of ligand-bound kinases showed that at least two hydrogen bonds between the ligand and the backbone protein atoms in the kinase hinge region are present in 90% of the complexes, with very little variability across targets. Closer inspection showed that when the two hydrogen bonds are present, one of three preferred hinge binding motifs is involved in 96.5% of the cases. Less than 10% of the false positives satisfied these two criteria in the minimized docking poses generated by our standard protocol. Ligand conformational artifacts were also shown to contribute to the occurrence of false positives in a number of cases. Application of this knowledge in the form of docking constraints and post-processing filters provided consistent improvements in virtual screening performance on all systems. The false-positive rates were significantly reduced and the enrichment factors increased by an average of twofold. On the basis of these results, a generalized two-step protocol for virtual screening on kinase targets is suggested. PMID- 16708366 TI - A BAC library for the goldfish Carassius auratus auratus (Cyprinidae, Cypriniformes). AB - A goldfish (Carassius auratus auratus) bacterial artificial chromosome genomic library (BAC library) was constructed from one aquarium-bred male specimen (tetraploid, 4n=100, genome size=3.52 pg/cell). The library consists of 128,352 positive clones with an average insert size of 150.4 kb, covering the genome 11 fold. All clones were spotted onto nylon filters and thus are available for screening of genomic regions of interest, such as candidate genes, gene families, or large-sized syntenic DNA regions of cyprinid species. Preliminary screens with two genes were conducted with hybridizing probes to the genes RAG1 and lgi1. RAG1 is a single-copy gene in zebrafish and is duplicated in C. a. auratus. We found a very close correlation between the number of positive BAC clones and the expected library coverage. Two copies of lgi1 were found in zebrafish. We have detected four different copies in C. a. auratus, not in the expected abundance, which indicates some variation in the coverage of the BAC library. The preliminary screens indicate that many duplicated genes that resulted from the ancient fish specific genome duplication persist in the tetraploid goldfish genome. Hence, the BAC library will provide a useful resource for the future work on comparative genomics, polyploidy, diploidization, and evolutionary genomics in fishes. PMID- 16708367 TI - Nerve-terminal and Schwann-cell response after nerve injury in the absence of nitric oxide. AB - Dystrophic muscles show alterations in the dystrophin-glycoprotein complex and a lack of neuronal nitric oxide (NO) synthase. In mdx mice, presynaptic expression of neuronal NO synthase is decreased, suggesting that presynaptic signaling may be altered in dystrophic muscle. In this study, we examined the nerve-terminal and Schwann-cell responses after a crush lesion in control and NO-deficient mice. Seven days after nerve crush, 24% of control neuromuscular junctions (n = 200) showed ultraterminal sprouts, whereas in NO-deficient mice this frequency was 28.5% (n = 217; P > 0.05 compared to controls; chi-square test). Schwann-cell response did not change in the absence of NO, after a nerve lesion of 7-day duration. Fourteen days after the lesion, nerve terminals sprouted and Schwann cells showed an extensive network of processes away from the synaptic site in controls. In the absence of NO, there was a dramatic decrease in nerve-terminal sprouting and Schwann-cell processes failed to extend away from the endplate. These results show that NO is involved in the nerve-terminal and Schwann-cell response to nerve injury. They also suggest that presynaptic molecular signaling may be impaired in dystrophic muscles, and this could influence the innervation and survival of newly formed myofibers generated by cell-mediated therapies. PMID- 16708369 TI - Cell-cycle regulation and mammalian gametogenesis: a lesson from the unexpected. AB - The progression of mammalian gametogenesis requires a precise balance between cell-cycle activities and elimination of defective gametogenic cells to ensure the perpetuation of species. Both spermatogonia and oogonia are stem cell populations committed to meiosis with the aim of generating haploid gametes for fertilization. At puberty, mitotically dividing spermatogonial cell cohorts maintain the ability of cell renewal and occupy niches in the seminiferous tubule. In contrast, mitotically dividing oogonial cell cohorts produced in the fetal ovary, are exclusively committed to meiosis and produce primordial follicles housing a primary oocyte surrounded by somatic follicular cells. A consistent physiological event during mammalian gametogenesis is the disposal of spermatogenic cells by apoptosis and ovarian follicles by atresia. Cyclin dependent kinases (Cdks) and their cyclin partners coordinate the activities of the cell cycle. An additional cell-cycle regulatory component is the centrosome. The centrosome harbors regulatory proteins controlling the normal progression of the cell cycle. Changes in individual centrosome proteins can lead to cell-cycle arrest and a decrease in the genomic protective function of p53 that promotes apoptosis. Disruption of cyclin A1, Cdk2, and Cdk4 expression in transgenic mice results in infertility and gonadal atrophy. Cdk-cyclin complexes interact with regulatory proteins, which may fine-tune the activities of the complex. One of the many regulatory proteins is p12, a 115 amino acid growth suppressor polypeptide designated p12(CDK2AP1), partner of Cdk2 and with binding affinity to DNA polymerase alpha/primase. Overexpression of p12 is associated with testicular and ovarian atrophy without affecting fertility. Ectopic expression of p12 was driven by the keratin 14 promoter. Keratin 14 is the pairing partner of keratin 5 and both keratins are expressed in testis. The efficiency of keratin promoters in driving ectopic gonadal gene expression, the association of gonadal atrophy with the ectopic expression of a Cdk2 regulatory protein and the centrosome, as a reservoir of cell-cycle regulatory proteins, open new experimental opportunities to address still lingering questions concerning cell differentiation and division during mammalian gametogenesis. PMID- 16708368 TI - Value of repeated measures of nerve conduction and quantitative sensory testing in a diabetic neuropathy trial. AB - Conduct of a large, multicenter trial of the aldose reductase inhibitor zenarestat provided data on the reproducibility of multiple electrophysiologic (nerve conduction studies, NCS) and quantitative sensory (QST) tests. Baseline and 12-month electrophysiologic data from approximately 1100 patients at multiple centers were available for analysis. Intersite variability contributed minimally to overall test variance. All NCS tests were highly reproducible. Cool thermal and vibration QST thresholds, as measured by CASE IV instrumentation, were also highly reproducible. Intersubject variance accounted for the majority of variance for all parameters measured. Repeating NCS and QST measures decreased sample sizes needed to show statistical significance. Consideration of these observations, particularly with regard to QST, should aid in the design of future clinical trials investigating neuropathy. PMID- 16708370 TI - Mutations within Wnt pathway genes in sporadic colorectal cancers and cell lines. AB - Wnt signaling pathway activation via mutation of genetic components, commonly adenomatous polyposis coli (APC), has a major role in colorectal cancer (CRC). Most components have not been assessed for mutation in sporadic CRC. We have analyzed AXIN2, CK1alpha, DKK1, GSK-3beta, SOX17, LRP6 and PPP2R1B, beta-catenin and APC in a collection of sporadic CRCs (n = 47) and CRC cell lines (CLs; n = 26). The CRC set was enriched for microsatellite unstable cancers (MSI+, 30%, 14/47). Somatic mutation was not found in CK1alpha, DKK1, LRP6, beta-catenin or GSK-3beta; but heterozygous frame-shift mutations, and an in-frame deletion mutation were detected in exon 7 of AXIN2 (CRCs, 11%, 5/47; CLs, 8%, 2/26). Our data refute a previous suggestion that a CRC-related mutational hot-spot occurred in the Huntington elongation A subunit TOR (HEAT) repeat 2 of PPP2R1B; this "hotspot" is, more likely, a rare germline polymorphism. An early investigation proposing a high mutational frequency in HEAT repeat 13 was not substantiated. A heterozygous SOX17 mutation (L194P) was also found in a cell line. APC gene mutations were identified in 64% (30/47) of cancers and 7% of these (2/30) had an additional mutation in another Wnt gene. Overall, 70% (33/47) of CRCs had a somatic mutation in a Wnt pathway gene. The number of tumors containing such a mutation was not significantly higher in MSI+ (57%, 8/14) compared to MSI- (76%, 25/33) cancers (p = 0.3, Fisher's exact test); APC mutation was significantly increased in the MSI- subgroup (p = 0.02, Fisher's exact test). Further, mutational screening of other Wnt pathway genes is warranted. PMID- 16708371 TI - Association of vitamin D receptor FokI polymorphism with prostate cancer risk, clinicopathological features and recurrence of prostate specific antigen after radical prostatectomy. AB - To investigate the effect of vitamin D receptor (VDR) FokI polymorphism on susceptibility to prostate cancer and the outcome of the disease in a Taiwanese population, we genotyped a total of 416 prostate cancer patients, 502 age-matched male controls and 189 non age-matched symptomatic benign prostatic hyperplasia. Although we did not find a significant association between VDR FokI genotypes and overall prostate cancer risk, we found that in men aged less than or equal to the median age of 73 years with VDR FokI F allele specifically had an increased risk of prostate cancer with a marginal significant trend (OR, 2.08; 95% CI, 1.00 4.34, p for trend = 0.056). The FF genotype was also highly associated with more aggressive prostate cancer (Gleason score 8-10) (OR, 2.47; 95% CI, 1.20-5.08) than did the Ff and ff genotypes. After adjusting other covariates, we found that in patients who had localized prostate cancer for which a radical prostatectomy was performed (n = 131), the VDR FokI FF genotype was associated with worse prostate-specific antigen (PSA) recurrence-free survival (hazard ratio = 3.25, 95% CI = 1.32-8.00, p = 0.010). Our findings suggest that the VDR FF genotype may increase the risk of early-onset prostate cancer and is associated with more aggressive disease. Furthermore, the VDR polymorphism could be used as a prognostic marker for localized prostate cancer after radical prostatectomy. PMID- 16708374 TI - Abstracts of the 46th Teratology Society Annual Meeting, 30th Annual Meeting of the Neurobehavioral Teratology Society, 19th International Conference of the Organization of Teratology Information Specialists, June 24-29, 2006, Tucson, Arizona, USA. PMID- 16708372 TI - Prevalence and determinants of human papillomavirus infection in men attending vasectomy clinics in Mexico. AB - Large studies of genital human papillomavirus (HPV) infection in men are few and mainly include high-risk groups. We interviewed 779 men who requested a vasectomy in 27 public clinics in 14 states of Mexico. Exfoliated cells were obtained from the scrotum, the shaft of the penis, the top of the penis including the coronal sulcus, the glans and the opening of the meatus. HPV testing was performed using biotinylated L1 consensus primers and reverse line blot. Unconditional logistic regression was used to estimate odds ratios (ORs) of being HPV-positive and corresponding 95% confidence intervals (CIs). The prevalence of any type of HPV was 8.7%. HPV positivity was highest among men below age 25 (13.6%), and lowest among men aged 40 years or older (6.0%). The most commonly found HPV types were, in decreasing order, HPV59, 51, 6, 16 and 58. Lifetime number of sexual partners was associated with HPV positivity (OR for > or = 4 vs. 1 partner = 3.7, 95% CI: 2.0-6.8), mainly on account of the strong association with number of occasional and sex-worker partners. Condom use with both regular (OR = 0.4, 95% CI: 0.1-1.0) and sex-worker (OR = 0.1, 95% CI: 0.0-0.3) partners and circumcision (OR = 0.2, 95% CI: 0.1-0.4) were inversely associated with HPV positivity. HPV prevalence in Mexican men was similar to the prevalence found in Mexican women of the same age groups. The association between HPV positivity and lifetime number of sexual partners in the present low-risk male population is one of the strongest ever reported in studies in men. Condom use and circumcision were associated with a strong reduction in HPV prevalence. PMID- 16708378 TI - Screening and chromatographic assessing of a novel IgG biomimetic ligand. AB - A novel biomimetic ligand, N-benzyloxycarbonyl-l-tyrosine (N-cbz-l-Tyr), was screened by a combination method of molecular docking and immobilized receptor technique. Then, N-cbz-l-Tyr was immobilized on Sepharose CL-4B to prepare a specific affinity adsorbent for immunoglobulin G (IgG). Scatchard analysis of the binding isotherm for IgG on the adsorbent gave an association constant (K(a)) of 4.91 x 10(6) m(-1) and a theoretical maximum adsorption capacity of 17.3 mg IgG/mL gel. IgG with a purity of 98% was separated from human plasma by this new affinity adsorbent. PMID- 16708379 TI - Determination of human plasma protein binding of baicalin by ultrafiltration and high-performance liquid chromatography. AB - A simple and selective HPLC assay was developed and utilized for determination of human plasma protein binding of baicalin. The method involved solid-phase extraction and reversed-phase chromatographic separation with a mobile phase of acetonitrile-0.02 mol/L phosphate buffer (pH 2.5; 25:75, v/v) and UV detection at 276 nm. The standard curve for baicalin was linear over the concentration range 0.1-20 microg/mL and the limit of detection was 0.02 microg/mL. The absolute recovery was greater than 76%. The intra-day and inter-day variations were less than 10%. Ultrafiltration technique was applied to determining the plasma protein binding of baicalin in human plasma. Results show the plasma protein binding of baicalin was in the range 86-92% over all the concentrations studied and the protein binding association constant was determined to be 1.21 x 10(5) L/mol at 4 degrees C. PMID- 16708380 TI - Food patterns and risk of breast cancer: A factor analysis study in Uruguay. AB - To generate broad eating patterns, which could explain more adequately the breast cancer etiology, we conducted an exploratory factor analysis in Montevideo, Uruguay. The study included 442 newly diagnosed and microscopically confirmed cases with breast cancer and 442 hospitalized controls, with non-neoplastic diseases. Factor analysis (principal components) was conducted in the control series, and as a result, 6 factors were extracted. These factors were labeled as traditional, healthy, western, stew, high-fat and drinker. The model explained 58.3% of the variance. After scoring the rotated factors, the relations between scores and breast cancer risk factors were analyzed by using Pearson correlation coefficients. After this step, the odds ratios of breast cancer for continuous scores of the rotated factors were carefully analyzed. The highest risk was directly associated with the western diet (OR 1.31, 95% CI 1.13-1.51), whereas the traditional (OR 0.77, 95% CI 0.64-0.93), healthy (OR 0.84, 95% CI 0.73-0.98) and stew (OR 0.83, 95% CI 0.71-0.98) diets were significantly protective. Women who reported a history of breast cancer among mother and sisters displayed strong elevations in risk for western (OR 2.03, 95% CI 1.11-3.72) and high-fat (OR 2.72, 95%CI 1.16-6.37) dietary patterns. This finding could suggest that gene-dietary interaction could play an important role in breast carcinogenesis. PMID- 16708381 TI - A highly specific real-time RT-PCR method for the quantitative determination of CK-19 mRNA positive cells in peripheral blood of patients with operable breast cancer. AB - The aim of the present study was to decrease the incidence of false positives and to better characterize marginally cytokeratin-19 (CK-19) mRNA positive peripheral blood samples from patients with early stage breast cancer. A new set of highly specific primers for CK-19, which avoids amplification of contaminating genomic DNA, was designed and evaluated to improve the specificity and sensitivity of the previously described methodology. The primers were specifically designed to avoid amplification of contaminating genomic DNA and CK-19 pseudogenes. The breast cancer cell line MCF-7 was used as positive control for the development and analytical evaluation of the assay, while peripheral blood samples from 62 healthy female individuals and 160 patients with early breast cancer were used for the evaluation of the sensitivity and specificity of the new primer pair. The novel designed primer pair was highly sensitive, as it detects up to 1 MCF-7 cell, and specific as none of the healthy individuals had detectable CK-19 mRNA positive cells in their peripheral blood. CK-19 mRNA positive cells were detected in 33 out of 160 (20.6%) patients with early breast cancer. Results obtained by the proposed optimized real-time RT-PCR protocol correlated well with those obtained in the same samples by our previously reported quantitative real-time RT PCR [concordance in 198/222 (89.2%), p = 0.0022, McNemar test]. The improved method eliminates the incidence of false positives and is highly sensitive and specific. The method could be used in a clinical setting in the near future for continuous monitoring and quantification of circulating epithelial cells in the peripheral blood of patients with operable breast cancer, provided that a quite larger number of clinical samples with a known follow-up will be analyzed. PMID- 16708382 TI - Epigenetic inactivation of the SFRP genes is associated with drinking, smoking and HPV in head and neck squamous cell carcinoma. AB - The soluble frizzled receptor protein (SFRP) family encodes antagonists of the WNT pathway, and silencing of these genes, through promoter hypermethylation, leads to constitutive WNT signaling. In bladder cancers, hypermethylation of the SFRP genes occurs more often in current and former smokers and is a strong predictor of poor patient survival. Hence, we examined methylation of these genes in another tobacco-related epithelial cancer, head and neck squamous cell carcinoma (HNSCC), to determine if the pattern of tobacco exposure again predicts the epigenetic alteration of these genes. Using methylation-specific PCR, the prevalence of methylation of SFRP1, SFRP2, SFRP4 and SFRP5 was 35, 32, 35 and 29%, respectively among 350 HNSCC cases. Promoter methylation of SFRP1 occurred more often in both heavy (OR 3.5, 95% CI 0.9-13.7) and light drinkers (OR 3.7, 95% CI 1.0-14.3) compared to nondrinkers. SFRP4 promoter methylation, on the other hand, occurred at a higher prevalence in never smokers and former smokers than in current smokers, and also was independently associated with HPV16 viral DNA. A joint effects model of SFRP4 promoter methylation demonstrated that smoking status and HPV virus significantly interacted (p < 0.04) such that never smokers with HPV16 had an OR of SFRP4 methylation of 9.0 (95% CI 2.1-38.6). These results suggest that epigenetic alterations of the SFRP genes are highly prevalent in HNSCC, and that the clonal selection for these alterations is complex and may be related to the carcinogenic exposures that are known risk factors for this disease. PMID- 16708383 TI - Prognostic significance of cyclin A in gastric cancer. AB - High level of cyclin A promotes carcinogenesis, and overexpression of cyclin A has been associated with poor prognosis of cancer patients. We validated the prognostic role of cyclin A in gastric cancer and evaluated its correlation with expression of an mRNA stability factor HuR. From 342 consecutive histologically confirmed gastric cancer patients were obtained 325 representative tissue specimens for cyclin A and 316 for HuR immunohistochemistry. Specimens were stained by cyclin A and HuR specific monoclonal antibodies. Nuclear immunostaining detected in > or =5% of the tumor cells was considered the cut-off for cyclin A positivity. Positive HuR immunoreactivity was scored as nuclear or cytoplasmic. Associations between scores, clinicopathological factors and survival were calculated by the chi2-test, Fisher's exact test, Kaplan-Meier test and Cox model. Cyclin A detected in the nuclei of cancer cells was positive in 55% (179 of 325) of the specimens; 40% (127 of 316) of the specimens had cytoplasmic and 88% (279 of 316) nuclear immunoreactivity of HuR. Cyclin A expression was an independent prognostic factor for poor survival. Cyclin A immunoreactivity was associated with old age, high stage, proximal location of the tumor, intestinal type, noncurative resection, advanced penetration depth and with nodal metastases but not distant metastases. Furthermore, cyclin A expression was associated with cytoplasmic HuR expression, whereas no association with nuclear HuR was evident. Cyclin A is an independent prognostic factor in gastric cancer, and one mechanism for its overexpression may depend on cytoplasmic localization of HuR. PMID- 16708384 TI - CYP24, the enzyme that catabolizes the antiproliferative agent vitamin D, is increased in lung cancer. AB - 1Alpha,25-dihydroxyvitamin D3 (1,25D3) displays potent antiproliferative activity in a variety of tumor model systems and is currently under investigation in clinical trials in cancer. Studies were initiated to explore its potential in nonsmall cell lung cancer (NSCLC), as effective approaches to the treatment of that disease are needed. In evaluating factors that may affect activity in NSCLC, the authors found that CYP24 (25-hydroxyvitamin D3-24-hydroxylase), the enzyme that catabolizes 1,25D3, is frequently expressed in NSCLC cell lines but not in the nontumorigenic bronchial epithelial cell line, Beas2B. CYP24 expression by RT PCR was also detected in 10/18 primary lung tumors but in only 1/11 normal lung tissue specimens. Tumor-specific CYP24 upregulation was confirmed at the protein level via immunoblot analysis of patient-matched normal lung tissue and lung tumor extracts. Enzymatically active CYP24 is expected to desensitize NSCLC cells to 1,25D3. The authors therefore implemented a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) assay for 1,25D3 and its CYP24-generated metabolites to determine whether NSCLC cells express active enzyme. Analysis of NSCLC cell cultures revealed time-dependent loss of 1,25D3 coincident with the appearance of CYP24-generated metabolites. MK-24(S)-S(O)(NH) Ph-1, a specific inhibitor of CYP24, slowed the loss of 1,25D3 and increased 1,25D3 half-life. Furthermore, combination of 1,25D3 with MK-24(S)-S(O)(NH)-Ph-1 resulted in a significant decrease in the concentration of 1,25D3 required to achieve maximum growth inhibition in NSCLC cells. These data suggest that increased CYP24 expression in lung tumors restricts 1,25D3 activity and support the preclinical evaluation of CYP24 inhibitors for lung cancer treatment. PMID- 16708385 TI - HPV16-E6 associated hTERT promoter acetylation is E6AP dependent, increased in later passage cells and enhanced by loss of p300. AB - The E6 oncoprotein from high-risk HPV types activates human telomerase reverse transcriptase (hTERT) transcription in human keratinocytes. Studies on how E6 regulates hTERT have implicated E-box or X-box elements in the hTERT promoter (Veldman et al., Proc Natl Acad Sci USA 2003;100:8211-14; Oh et al., J Virol 2001;75:5559-66; Gewin et al., Genes Dev 2004;18:2269-82), but the mechanism of activation by E6 is still controversial and not well defined. Here, we demonstrate that induction of both hTERT expression and telomerase activity by HPV-16 E6 in early passage keratinocytes is associated with acetylation of histone H3 at the hTERT promoter, is dependent on the E6 associated protein (E6AP) and is not exclusively reliant on E-box or X-box elements. Further increases in histone acetylation of the hTERT promoter and hTERT transcriptional activity in E6 expressing cells that had been passaged extensively in culture were found to occur only with the endogenous promoter and not with an exogenously introduced hTERT promoter construct. Telomerase activity at both early and late passages, however, was dependent on E6AP expression, implying a continued reliance on E6 function for telomerase activity. Our results demonstrate that E6 induces hTERT promoter acetylation, but that further increases in telomerase activity and histone acetylation in later passage E6 expressing cells are independent of E6 activation of the core hTERT promoter. We also provide evidence that the transcription factor p300 is a potential repressor of telomerase activation and histone acetylation in the context of E6 expression. These studies give insight into how immortalization by HPV results in upregulation of hTERT and furthers our understanding of how telomerase is activated during the process of malignant transformation. PMID- 16708386 TI - Nas transgenic mouse line allows visualization of Notch pathway activity in vivo. AB - The Notch signaling pathway plays multiple and important roles in mammals. However, several aspects of its action, in particular, the precise mapping of its sites of activity, remain unclear. To address this issue, we generated a transgenic line carrying a construct consisting of a nls-lacZ reporter gene under the control of a minimal promoter and multiple RBP-Jkappa binding sites. Here we show that this transgenic line, which we termed NAS (for Notch Activity Sensor), displays an expression profile that is consistent with current knowledge on Notch activity sites in mice, even though it may not report on all these sites. Moreover, we observe that NAS transgene expression is abolished in a RBP-Jkappa deficient background, indicating that it indeed requires Notch/RBP-Jkappa signaling pathway activity. Thus, the NAS transgenic line constitutes a valuable and versatile tool to gain further insights into the complex and various functions of the Notch signaling pathway. PMID- 16708387 TI - Variation in retinitis pigmentosa-11 (PRPF31 or RP11) gene expression between symptomatic and asymptomatic patients with dominant RP11 mutations. AB - Dominant mutations in the mRNA splicing factor gene PRPF31 (RP11) cause retinitis pigmentosa with reduced penetrance. We studied the expression of RP11 in lymphoblast cell lines from 10 patients, including three who were clinically asymptomatic, with six distinct RP11 mutations. Five of the six mutations were characterized and all five created premature nonsense codons or eliminated the normal initiation codon. Semiquantitative RT-PCR indicated that an average of only 17% of the RP11 mRNA was derived from the mutant allele, likely because the mutant mRNA transcripts were degraded by nonsense-mediated decay. Gene expression levels were measured by Affymetrix and CodeLink microarrays and, for RP11 transcripts, also by real-time PCR. Combined wild-type-plus-mutant RP11 mRNA expression from symptomatic patients was 52 to 77% of that in controls (p < or = 0.0005). Clinically asymptomatic carriers had levels of RP11 mRNA similar to controls and 29-42% higher than in clinically affected patients (0.0001 120 days). The combined therapy of B16/CCL21-Te-Fc plus anti-4-1BB MAbs further enhanced the immune response, resulting in 75% of tumor-bearing mice achieved long-term survival (> 120 days) in subcutaneous model and few lung nodules in pulmonary metastasis model. Rechallenge experiment showed that a persistent memory response was successfully induced by the combined therapy. In vivo depletion of lymphocytes indicated that CD8+ T cells were essential in the antitumor activity induced by B16/CCL21-Te-Fc plus anti-4-1BB MAbs, whereas NK cells and CD4+ T cells played substantial roles. The CTL activity induced by pCCL21-Te-Fc-transfected PBMCs specifically lysed a variety of human leukocyte antigen-matched and hTERT-positive human tumor cells, suggesting pCCL21-Te-Fc could serve as a vaccine against most human cancers. PMID- 16708390 TI - Adenovirus-mediated tBid overexpression results in therapeutic effects on p53 resistant hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide with a very high mortality. Because the success of the conventional therapies is limited, gene therapy may represent an alternative for HCC management. Our earlier study has shown that Bid plays a role in the development of HCC. The aim of our study is to evaluate the possibility of using truncated Bid (tBid) as a novel therapy for HCC treatment. Two HCC cell lines, Hep3B and PLC/PRF/5, were used in the experiment. Hep3B was a p53-resistant while PLC/PRF/5 a p53 sensitive. A recombinant adenovirus-Ad/AFPtBid, which contained a tBid gene driven by an alpha-fetoprotein (AFP) promoter, was constructed. Both Hep3B and PLC/PRF/5 cells infected with Ad/AFPtBid showed a significant decrease in cell viability. The decrease in cell viability by Ad/AFPtBid resulted from apoptosis of HCC cells, evident by enhanced activity of caspases and increased release of cytochrome c. In vivo experiment was performed by the intratumor injection of Ad/AFPtBid in nude mice inoculated with Hep3B. Ad/AFPtBid injection significantly inhibited tumor growth, and tumor tissues showed a marked increase in TUNEL positive cells. Our experiment also demonstrated that Ad/AFPtBid only targeted AFP-producing cells but not those non-AFP producing cells. In conclusion, these results indicate that the introduction of Ad/AFPtBid can not only significantly but specifically kill HCC cells that produce AFP. The cell death induced by Ad/AFPtBid in HCC cells is via an apoptotic pathway that can be independent of p53 status. PMID- 16708391 TI - Mobility and integration sites of a murine C57BL/6 melanoma endogenous retrovirus involved in tumor progression in vivo. AB - Tumor development is a multistep process in which both genetic and epigenetic events cooperate for the emergence of a malignant clone with metastatic properties. The possibility that endogenous retroviruses promote the expansion of a neoplastic clone by subverting immunosurveillance has been proposed and recently demonstrated in the case of the B16 murine melanoma, which spontaneously express the melanoma-associated retrovirus (MelARV). Indeed, knocking down, by RNA interference, this endogenous retrovirus resulted in the rejection of the tumor cells in immunocompetent mice, without any alteration of their transformed phenotype. Here, we characterize the MelARV proviruses present in the B16 melanoma. Complete sequencing of the viral genomic RNA and characterization of the integration sites within both the B16 tumor cells and a subline selected in vivo for increased metastatic activity disclosed mobility of the element with new proviral insertions targeting critical genes and altering their transcriptional profile. The results show that MelARV can act both at the genetic level, inducing mutations by insertion, and at the epigenetic level, promoting immunosuppression of the host. These properties may as well be relevant to human tumors, such as germline tumors and melanoma, where endogenous retroviruses are active. PMID- 16708392 TI - Helicobacter pylori infection is not associated with increased risk of colorectal polyps in Taiwanese. PMID- 16708393 TI - Cooperation and competition between the dorsal hippocampus and lateral amygdala in spatial discrimination learning. AB - The conditioned cue preference (CCP) was used to study how rats discriminate between adjacent arms on a radial maze. Chai and White (Behav Neurosci 2004, 118:770-784) showed that an intact dorsal hippocampus is required to learn this discrimination and that an amygdala-based conditioned approach response that produces an equal tendency to enter both arms is simultaneously acquired. In the present experiments, rats were preexposed to the maze with no food and trained by alternately confining them at the ends of two adjacent arms, one that contained food and one that did not. When given a choice between these arms with no food present, the rats spent more time on their food-paired arms, suggesting they had learned to discriminate their locations. Temporary inactivation of the dorsal hippocampus with muscimol during confinement on the food-paired arm had no effect on the discrimination, but inactivation while on the no-food arm impaired it. This pattern of effects was reversed in rats with amygdala lesions (inactivation on the food-paired arm impaired, but inactivation on the no-food arm had no effect on the discrimination), showing that hippocampus-based and amygdala-based learning interact to influence the behavior of normal rats in this situation. The dorsal hippocampus learns about locations that contain food and about locations that do not contain food. The amygdala-based tendency to enter the food-paired arm cooperates with hippocampus-based foraging for food on the food-paired, but the amygdala-based tendency to enter the no-food arm competes with hippocampus based learning about the absence of food on that arm. PMID- 16708395 TI - Estimation of phospholipophilicity of 1-[3-(arylpiperazin-1-yl)-propyl] pyrrolidin-2-one derivatives on immobilized artificial membrane stationary phase and its correlation with biological data. AB - A molecule library containing 42 1-[3-(arylpiperazin-1-yl)-propyl]-pyrrolidin-2 one derivatives has been designed and synthesized. The phospholipophilicity of the obtained compounds has been determined using immobilized artificial membrane high-performance liquid chromatography (IAM-HPLC). The performed analysis allowed the calculation of log k(we) values for each of the tested compounds. Experimental phospholipophilicity data (log k(we)) has been compared with the affinity of the tested compounds to alpha(2)-adrenoceptors. Performed quantitative structure-activity relationship studies indicated that, for the tested compounds, there are dependences between affinity for alpha(2) adrenoceptors and their log k(we) values. The obtained results confirmed that the applied chromatographic IAM-HPLC method could be useful in fast characterization of the phosholipophilicity of structurally closely related compounds as well as for larger series of compounds, such as drug candidates. It could also be used as a tool for further research into this group of compounds. PMID- 16708394 TI - Tg(Afp-GFP) expression marks primitive and definitive endoderm lineages during mouse development. AB - Alpha-fetoprotein (Afp) is the most abundant serum protein in the developing embryo. It is secreted by the visceral endoderm, its derivative yolk sac endoderm, fetal liver hepatocytes, and the developing gut epithelium. The abundance of this protein suggested that Afp gene regulatory elements might serve to effectively drive reporter gene expression in developing endodermal tissues. To this end, we generated transgenic mouse lines Tg(Afp-GFP) using an Afp promoter/enhancer to drive expression of green fluorescent protein (GFP). Bright GFP fluorescence allowed the visualization, in real time, of visceral endoderm, yolk sac endoderm, fetal liver hepatocytes, and the epithelium of the gut and pancreas. Comparison of the localization of green fluorescence with that of endogenous Afp transcripts and protein indicated that the regulatory elements used to generate these mouse lines directed transgene expression in what appeared to be all Afp-expressing cells of the embryo, but only in a subset of fetal liver cells. The bright GFP signal permitted flow cytometric analysis of fetal liver hepatocytes. These mice represent a valuable resource for live imaging as well as identification, quantitation, and isolation of cells from the primitive and definitive endoderm lineages of the developing mouse embryo. PMID- 16708396 TI - Identification of the commonest cystic fibrosis transmembrane regulator gene DeltaF508 mutation: evaluation of PCR--single-strand conformational polymorphism and polyacrylamide gel electrophoresis. AB - In the present study we investigated whether single-strand conformational polymorphism (SSCP) and polyacrylamide gel electrophoresis (PAGE) could be used for the identification of the CFTR DeltaF508 gene mutation, which is commonest in the Greek population. Using DNA from patients carrying this mutation, the appropriate 98 bp region of the CFTR gene was amplified by PCR and the reaction products were analysed by non-radioactive SSCP-electrophoresis using silver staining for band visualization and non-denaturating PAGE to confirm the results. SSCP electrophoretic analysis has been optimized for several parameters in order to achieve the best resolution. Single-strand DNA fragments gave a reproducible pattern of bands, characteristic for the particular mutation. Comparison of the obtain patterns with control samples allowed the detection of the DeltaF508 mutation in the patients studied by SSCP assay and these results were confirmed by the independent method of PAGE. Although SSCP and PAGE can be used for detection of this mutation, PAGE resulted in more distinct patterns than SSCP. It is, therefore, proposed that PAGE can be reliably used for the detection and identification of such a mutation in patients provided that suitable controls are available. The applicability of PAGE to identification of the mutation in carriers, particularly useful for population screening, is also discussed. PMID- 16708397 TI - Integration of CD28 and CTLA-4 function results in differential responses of T cells to CD80 and CD86. AB - CD80 and CD86 have the capacity to either stimulate or inhibit T cell responses through their receptors CD28 and cytotoxic T lymphocyte-associated antigen 4 (CTLA-4). Blockade of CD80 and CD86 in autoimmune disease settings has revealed distinct outcomes, yet the differential functions of CD80 and CD86 are still unclear. We have studied the ability of individual ligands to stimulate primary responses in human CD4(+) T cells. Our data reveal both quantitative and qualitative differences between the ligands. Both CD80 and CD86 demonstrated the capacity to costimulate T cell proliferation. However, CD80 committed a greater number of T cells to divide with faster kinetics, consistent with it being a superior ligand for CD28. Once cell division had been initiated, all T cells undergoing cell division expressed CTLA-4, irrespective of whether CD80 or CD86 costimulation was used. However, only in the presence of CD80 was evidence of CTLA-4 engagement and inhibitory function observed. Finally, differences between CD80 and CD86 costimulation extended to the T cell phenotype, in particular the levels of CD40 ligand expression. PMID- 16708398 TI - Effects of dexamethazone on LPS-induced activationand migration of mouse dendritic cells revealed by a genome-wide transcriptional analysis. AB - While lipopolysaccharides (LPS) induce dendritic cell (DC) maturation and migration to lymph nodes, glucocorticoids such as dexamethazone (Dex) have a profound suppressive effect on immune response. The mechanisms that might control this suppressive effect of Dex have been extensively investigated in lymphocytes as possible targets. Much less is known on the effects of Dex on DC, although they are recognized to regulate immunity. To get insights into possible combined effects of Dex and LPS on DC functions, we have undertaken a genome-wide analysis of differentially expressed genes of DC treated with Dex alone, LPS alone, or both, using high-density oligonucleotide microarrays. Hierarchical clustering and principal component analysis (PCA) agreed in identifying 24 h as the time point that best discriminated the three treatments. Among the counteracting effects we have observed an inhibition of Dex on the LPS-induced up-regulation of the chemokine receptor CCR7. In vivo, Dex treatment blocked the LPS-induced migration of DC, which lost their ability to reach the draining lymph nodes. In addition, we observed a synergistic effect of Dex and LPS on the expression of the secreted lipocalin 24p3, which has been reported to induce apoptosis in T cells and thus may be related to immune suppression. PMID- 16708399 TI - Efficient induction of antitumor T cell immunity by exosomes derived from heat shocked lymphoma cells. AB - Exosomes secreted by tumor cells could serve as a promising immunotherapeutic tumor vaccine. Heat shock proteins (HSP) induced in tumor cells by heat shock are molecular chaperones with potent adjuvant activity in the induction of antigen specific T cell responses. To improve exosome-based tumor vaccines, we have investigated the efficacy of exosomes derived from heat-shocked mouse B lymphoma cells (HS-Exo) in the induction of antitumor immune responses. We found that HS Exo, compared with control exosomes derived from the same cells (Exo), contain more HSP60 and HSP90 and increased amounts of molecules involved in immunogenicity including MHC class I, MHC class II, CD40, CD86, RANTES and IL 1beta. Furthermore, HS-Exo induce both phenotypic and functional maturation of dendritic cells more efficiently. HS-Exo immunization activates T cell responses more potently. Importantly, HS-Exo induce dramatically increased antitumor immune responses compared to control exosomes from the same cells in prophylaxis and therapeutic in vivo lymphoma models. We further demonstrate that CD8(+) T cells are the predominant T cell subset responsible for the antitumor effect of HS-Exo and that CD4(+) T cells are necessary in the induction phase of tumor rejection in a prophylaxis model. These findings provide a novel strategy to improve the efficacy of exosome-based tumor vaccines. PMID- 16708400 TI - Evidence that a significant number of naive T cells enter non-lymphoid organs as part of a normal migratory pathway. AB - Only activated and effector memory T cells are thought to access non-lymphoid tissues. In contrast, naive T cells are thought to circulate only between the blood, lymph and secondary lymphoid organs. We examined the phenotype of endogenous T cells in various non-lymphoid organs and showed that a subset of cells exhibited an apparently naive phenotype and were functionally inactive. FTY720 treatment selectively depleted this population from the non-lymphoid tissues. In addition, RAG-deficient TCR transgenic CD4 and CD8 T cells were present in non-lymphoid tissues in bone marrow chimeric mice and in situ imaging analysis revealed their location in the parenchymal tissues. Moreover, migration of TCR transgenic T cells to non-lymphoid tissues after adoptive transfer was pertussis-toxin resistant. Overall, the results suggest that naive T cells may circulate through non-lymphoid tissues as part of their normal migratory pathway. PMID- 16708402 TI - Functional impairment of cytotoxic T cells in the lung airways following respiratory virus infections. AB - We investigated the differentiation phenotype and function of virus-specific and non-specific CTL that were recruited to the lung parenchyma and the bronchoalveolar space after respiratory virus infections. Soon after virus elimination, we observed functional impairment of CTL isolated from the airways in their ability to produce IFN-gamma and TNF-alpha and to lyse target cells. Impaired cytotoxicity was due to a reduced content of granzyme B and a reduced ability to mobilize lytic granules. This impairment in effector functions (a) was largely restricted to CTL in the lung airways, (b) affected both CTL specific for the infecting virus as well as those that were recruited non-specifically to the inflamed lung, (c) was independent of contact between CTL and their specific viral antigen, (d) was not restricted to terminally differentiated CTL but also affected resting memory CTL and (e) could be elicited by both respiratory syncytial virus and influenza virus and thus seemed to be largely independent of the infecting virus. These observations suggest that functional impairment of antiviral T cells in the lung is not the consequence of a viral escape strategy. It may rather result from the particular milieu in the bronchoalveolar space and reflect a host mechanism to prevent excessive pulmonary inflammation. PMID- 16708401 TI - Alloiococcus otitidis is a ligand for collectins and Toll-like receptor 2, and its phagocytosis is enhanced by collectins. AB - Alloiococcus otitidis has been found to be associated with otitis media with effusion. In this study we investigated whether TLR2 and collectins, surfactant protein A (SP-A) and mannose-binding lectin (MBL), interacted with A. otitidis. Both SP-A and MBL bound to A. otitidis in a Ca(2+)-dependent manner. A. otitidis induced IL-8 secretion from U937 cells and NF-kappaB activation in TLR2 transfected HEK293 cells. However, the cells transfected with the mutant TLR2(P681H) did not respond to A. otitidis. In addition, A. otitidis co sedimented a recombinant soluble form of the extracellular TLR2 domain, indicating direct binding of the bacterium to TLR2. SP-A and MBL augmented the phagocytosis of A. otitidis by J774A.1 cells. The collectin-stimulated phagocytosis of A. otitidis was significantly attenuated when fucoidan and polyinosinic acid were co-incubated. Immunoblotting analysis revealed that MBL was present in the middle ear effusion from patients with otitis media. These results demonstrate that A. otitidis is a ligand for the collectins and TLR2, and that the collectins enhance the phagocytosis of A. otitidis by macrophages, suggesting important roles of the collectins and TLR2 in the innate immunity of the middle ear against A. otitidis infection. PMID- 16708403 TI - BLIMP-1 is a target of cellular stress and downstream of the unfolded protein response. AB - B lymphocyte-induced maturation protein-1 (BLIMP-1) acts during differentiation of B cells and monocytes, but was originally identified as a repressor of the IFN beta promoter induced during viral infection. A central regulator of the intracellular response to viral infection is the interferon-inducible double stranded RNA activated protein kinase (PKR). PKR belongs to a family of kinases that phosphorylate the eukaryotic translation initiation factor 2-alpha (eIF2alpha) and activate common downstream signaling pathways. PERK, the endoplasmic reticulum resident PKR-homologue, is activated during the unfolded protein response (UPR), a stress response involved in both macrophage activation and terminal B-cell differentiation. This suggested that BLIMP-1 might be a target of stress responses involving PERK. We demonstrate that BLIMP-1 is rapidly up-regulated during the UPR in human myeloid and B-cell lines. This response is conserved in murine B-cells and murine macrophages, in which mimics of physiological stress and classical activation stimuli also induce Blimp-1. During the UPR, BLIMP-1 mRNA is induced at the level of transcription. This response is dependent on an intact PERK signaling pathway, independent of new protein synthesis and blocked by an inhibitor of NF-kappaB. Our data provide evidence for a novel pathway linking cellular stress to BLIMP-1, a regulator of differentiation in macrophages and B cells. PMID- 16708404 TI - Colitogenic and non-colitogenic commensal bacteria differentially trigger DC maturation and Th cell polarization: an important role for IL-6. AB - We investigated whether commensal bacteria modulate activation and maturation of bone marrow-derived DC and their ability to prime CD4(+) T cells. We used Escherichia coli mpk, which induces colitis in gnotobiotic IL-2-deficient (IL-2( /-)) mice, and Bacteroides vulgatus mpk, which prevents E. coli-induced colitis. Stimulation of DC with E. coli induced TNF-alpha, IL-12 and IL-6 secretion and expression of activation markers. Moreover, stimulation of DC with E. coli increased T cell activation and led to Th1 polarization. Stimulation with B. vulgatus led only to secretion of IL-6, and DC were driven into a semi-mature state with low expression of activation markers and did not promote Th1 responses. B. vulgatus-induced semi-mature DC were non-responsive to stimulation by E. coli in terms of maturation, T cell priming and TNF-alpha but not IL-6 production. The non-responsiveness of B. vulgatus-stimulated DC was abrogated by addition of anti-IL-6 mAb or mimicked with rIL-6. These data suggest that B. vulgatus-induced IL-6 drives DC into a semi-mature state in which they are non responsive to proinflammatory activation by E. coli. This in vitro mechanism might contribute to the prevention of E. coli-triggered colitis development by B. vulgatus in vivo; high IL-6 mRNA expression was consistently found in B. vulgatus colonized or B. vulgatus/E. coli co-colonized IL-2(-/-) mice and was associated with absence of colitis. PMID- 16708406 TI - The hidden truth about gene expression in Tregs: is it what you don't see that counts? AB - A population of IL-10 secreting regulatory T cells (PI-Tregs) can be induced in a mouse TCR transgenic model of experimental autoimmune encephalomyelitis (EAE) by intranasal administration of the N-terminal peptide of myelin basic protein. A paper in this issue of the European Journal of Immunology identifies patterns of gene expression that distinguish PI-Treg from naive T cells after activation in vivo. PI-Tregs are anergic and the expression of egr2 may be involved in regulation of the cell cycle. The surprising expression of the Th1 determining gene T-bet in PI-Tregs may be functionally relevant as transfection of T-bet into a hybridoma induced some genes associated with regulatory T cells, such as tlr2 and gzmb. In this commentary, PI-Tregs are considered in comparison to other Tregs, such as the natural CD4(+)CD25(+) and Tr1 cells, and there seems to be considerable overlap in gene expression between all three Treg populations, but also with both Th1 and Th2 cells. It is suggested that regulatory activity may be a common feature of all activated T cells and that a defining principle of a Treg is the lack of effector functions due to a partial or incomplete differentiation to either Th1 or Th2. PMID- 16708407 TI - Pharmacological activity of three commercial Hypericum perforatum preparations in mice. AB - St John's Wort (Hypericum perforatum) is the main herbal species used to treat depression. The products available on the pharmaceutical and dietary supplement markets are obtained by a variety of preparation processes and their pharmacological effects may differ significantly. The purpose of this study therefore was to investigate the effect of different St John's Wort commercial preparations available on the French market. Only one preparation gave significant results in the forced swimming test. PMID- 16708408 TI - Bulk agent Plantago ovata after Milligan-Morgan hemorrhoidectomy with Ligasure. AB - The aim of this study was to determine usefulness of the bulk agent Plantago ovata in reducing postoperative pain and tenesmus after open hemorrhoidectomy (Milligan-Morgan with Ligasure). Ninety-eight patients were randomized into two groups of 49 patients each. In both groups Milligan-Morgan open hemorrhoidectomy with Ligasure was performed. The first group received postoperatively two sachets daily of 3.26 g of the bulk agent, Plantago ovata, for 20 days. The control group was treated postoperatively with glycerin oil. There was no statistically significant difference in age, gender distribution and hemorrhoid grading, between the two groups. The pain score after first defecation (p < 0.001) and after 10 days (p < 0.01) and the global pain score (p < 0.001) was statistically significantly lower in the group treated with Plantago ovata, while there was no statistically significant difference in the pain level after 20 days (p > 0.05). The hospital stay was statistically significantly shorter in the group receiving Plantago ovata (2.6 +/- 0.6 vs 3.9 +/- 0.7 days, p < 0.001). The incidence of tenesmus was higher in the control group (40.8% vs 10.2%, p < 0.01). Treating patients with Plantago ovata after open hemorrhoidectomy, reduces pain, tenesmus rate and shortens postoperative hospital stay. PMID- 16708405 TI - Persistent antigenic stimulation alters the transcription program in T cells, resulting in antigen-specific tolerance. AB - Repetitive antigen stimulation induces peripheral T cell tolerance in vivo. It is not known, however, whether multiple stimulations merely suppress T cell activation or, alternatively, change the transcriptional program to a distinct, tolerant state. In this study, we have discovered that STAT3 and STAT5 were activated in response to antigen stimulation in vivo, in marked contrast to the suppression of AP-1, NF-kappaB and NFAT. In addition, a number of transcription factors were induced in tolerant T cells following antigen challenge in vivo, including T-bet, Irf-1 and Egr-2. The altered transcription program in tolerant cells associates closely with the suppression of cell cycle progression and IL-2 production, as well as with the induction of IL-10. Studies of T-bet and Egr-2 show that the function of T-bet in peptide treatment-induced regulatory T cells is not associated with Th1 differentiation, but correlates with the suppression of IL-2, whereas expression of Egr-2 led to an up-regulation of the cell cycle inhibitors p21(cip1) and p27(kip). Our results demonstrate a balanced transcription program regulated by different transcription factors for T cell activation and/or tolerance during antigen-induced T cell responses. Persistent antigen stimulation can induce T cell tolerance by changing the balance of transcription factors. PMID- 16708409 TI - Rapid identification of potent nonpeptidic serine protease inhibitors. PMID- 16708411 TI - Energetic nitrogen-rich salts and ionic liquids. AB - Energetic salts offer many advantages over conventional energetic molecular compounds. The use of nitrogen containing anions and cations contributes to high heats of formations and high densities. Their low carbon and hydrogen content gives rise to a good oxygen balance. The decomposition of these compounds is predominantly through the generation of dinitrogen which makes them very promising candidates for highly energetic materials for industrial or military applications. PMID- 16708413 TI - Highly enantioselective aza-Baylis-Hillman reaction in a chiral reaction medium. PMID- 16708414 TI - Development of a common fully stereocontrolled access to the medicinally important and promising prostacyclin analogues iloprost, 3-oxa-iloprost and cicaprost. AB - We describe new fully stereocontrolled syntheses of the prostacyclin analogues iloprost (2), the most active component of the drugs Ilomedin and Ventavis, and 3 oxa-iloprost (3), a derivative that is expected to have a significantly higher metabolic stability than 2 perhaps allowing an oral application. The syntheses are based on the same strategy and chiral bicyclic building block as used in the synthesis of cicaprost (4), the third most potent analogue that exhibits, besides prostacyclin-like activities, antimetastatic activities. Reaction of the enantiopure C6-C13 bicyclic aldehyde 17 with Cl(3)CCOOH/Cl(3)CCOONa afforded trichlorocarbinol 24 which was converted via mesylate 25 to the C6-C14 bicyclic alkyne 9. The palladium-catalysed hydrostannylation of alkyne 9 gave with high regio- and stereoselectivity the alkenylstannane 26, Sn/Li exchange of which afforded the E-configured alkenyllithium derivative 8. Coupling of the C6-C14 building block 8 with the enantiopure C15-C20 building block, the N-methoxyamide 7, gave the C6-C20 bicyclic ketone 6 in high yield without epimerisation at C16. The configuration at C15 of iloprost (2) and 3-oxa-iloprost (3) was established through a highly diastereoselective reduction of ketone 6 with catecholborane and the chiral oxazaborolidine 28 which furnished alcohol (15S)-29. The highly stereoselective conversions of alcohol (15S)-29 to iloprost (2) and 3-oxa iloprost (3), which include as key stereoselective steps an olefination with a chiral phosphonoacetate and a copper-mediated allylic alkylation, have already been described. PMID- 16708416 TI - The influence of fetal position on amniotic fluid index and single deepest pocket. AB - OBJECTIVE: To evaluate the effect of fetal position on measurement of amniotic fluid index (AFI) and of the single deepest pocket (SDP). METHODS: This was a prospective observational study, in a university obstetric unit, of women with an uncomplicated singleton pregnancy with longitudinal lie and cephalic presentation at or beyond 28 weeks of gestation. AFI was calculated and SDP measured and the fetal position was characterized in terms of three parameters. These were: the side of the maternal abdomen on which the fetus lay; a numerical representation (laterality score) of the distance that the fetus was from the sagittal midline plane of the maternal abdomen; the orientation of the fetal trunk (ventral anterior, lateral or posterior). RESULTS: Eighty-one women were recruited into the study. There was a significant relationship between the laterality score and the AFI (P = 0.005) but not the SDP (P = 0.23): AFI was on average 4.35 cm higher in fetuses lying centrally compared with those lying laterally inside the uterus. There was no significant difference for either SDP (P = 0.8) or AFI (P = 0.3) between fetuses lying on the right or the left side of the maternal abdomen. Similarly, there was no significant difference in SDP (P = 0.9) or AFI (P = 1.0) for the different orientations of the fetal trunk. CONCLUSION: Fetal position affects the measurement of AFI but not that of SDP. Therefore, SDP may be a more consistent parameter for the estimation of amniotic fluid volume. PMID- 16708417 TI - Vision restoration therapy and raising red flags too early. PMID- 16708418 TI - Blurred vision and dizziness. Ocular flutter. PMID- 16708419 TI - Polypropylene mesh for hernia repair following transplantation. PMID- 16708420 TI - To biopsy or not to biopsy? PMID- 16708421 TI - Acute cellular rejection rates in living-donor and deceased-donor liver transplantation. PMID- 16708422 TI - Counterpoint: Heat-induced membrane depolarization of hypothalamic neurons: an unlikely mechanism of central thermosensitivity. PMID- 16708423 TI - Statement on persistent diarrhea in the returned traveller. An Advisory Committee Statement (ACS). PMID- 16708424 TI - Ultraviolet radiation increases the toxicity of pyrene, 1-aminopyrene and 1 hydroxypyrene to human keratinocytes. AB - Over the past several years, a great deal of interest has been focused on the harmful effects of ultraviolet (UV) radiation to human skin. UV light has been implicated in aging, sunburn and skin cancer. Few studies, however, have been done to determine the effects that UV light, in conjunction with other environmental contaminants, may have on human skin. Polycyclic Aromatic Hydrocarbons (PAHs) are a class of compounds that have been reported to be toxic, mutagenic and carcinogenic to many eukaryotic organisms. UV light is also known to increase the toxicity of PAHs through photo-activation and photo-modification. The purpose of this study was to assess the effects of UV-A irradiated pyrene (Pyr), 1-aminopyrene (1-AP) and 1-hydroxypyrene (1-HP) on human keratinocytes, the skin primary site of UV irradiated PAH exposure. Our findings indicate that simultaneous treatment of human keratinocyte cell line, HaCaT, with 1.0 microg/ml pyrene, 1-AP or 1-HP and 3.9 J/cm2/min UV-A light resulted in significant inhibition of cell proliferation. Approximately 100% of the cells died in the case of UV-A irradiated 1-AP and 1-HP. In the case of UV-A irradiated pyrene, more than 70% of the cells died, indicating that UV-A is able to transform these PAHs into more harmful intermediates. PMID- 16708426 TI - Proceedings of the 17th and 18th Nordic Committee for Veterinary Scientific Cooperation (NKVet) Symposia. PMID- 16708425 TI - Effects of pristane on cytochrome P450 isozyme expression in rat tissues. AB - Chemical carcinogenesis studies are powerful tools to obtain information on potential mechanisms of chemical factors for malignancies. In this study Western blot analyses, using monoclonal antibodies specific for three different cytochrome P450 (CYP) isozymes (CYP1A1, CYP1A2 and CYP2B), were employed to examine the effect(s) of 3-methylcholanthrene and/or pristane (2,6,10,14 tetramethylpentadecane) on the basal and inducible levels of expression of CYP proteins within Copenhagen rat tissues. Pristane exposure led to tissue specific differences in the CYP isozymes expressed and elicited increased CYP protein expression over 3-methylcholanthrene induced levels in microsomes isolated from liver, Peyer's Patches, and thymus. Within the context of the chemical carcinogenesis model employed in this study, these observations correlated with the induction of B-cell malignancies by low doses of 3-methylcholanthrene and of thymic lymphomas by a high 3-methylcholanthrene dose. The data suggest that pristane treatment affects CYP isozyme expression. This pristane-mediated effect clearly could be a contributing factor in the chemical carcinogenesis of the previously observed lymphoid malignancies, and a possible basis for the tumor enhancing effects of pristane. PMID- 16708427 TI - Psychotropic medication in the criminal trial process: the constitutional and therapeutic implications of Riggins v. Nevada. PMID- 16708428 TI - The effect of firearm deaths on life expectancy and insurance premiums in the United States. AB - Despite recent gains, the U.S. remains behind most other affluent countries in life expectancy. Even within the U.S., the gap between the life expectancies of Caucasians and African-Americans remains significant. At the same time, firearm deaths in the U.S. far exceed peer nations, and disproportionately affect African American males. In this Issue Brief, Dr. Lemaire explores whether deaths from firearms explain some of these international and racial disparities in life expectancy. He uses actuarial techniques to calculate the "cost" of firearm deaths in the U.S., both in terms of reduced life expectancy and increased life insurance premiums. PMID- 16708429 TI - Understanding and improving hospice enrollment. AB - Hospice is considered the "gold standard" for end-of-life care, providing dying patients and their families access to a broad array of services across settings. Despite its comprehensive approach, hospice care remains underutilized; many patients who might benefit from hospice do not enroll, or enroll only in the last days of life. This Issue Brief summarizes a series of studies that shed light on the decision making process about hospice, and describes a simple, effective way to improve referrals to hospice among nursing home residents. PMID- 16708430 TI - Preparing for the next natural disaster: learning from Katrina. AB - Hurricane Katrina exposed the nation's ongoing vulnerability to large-scale losses from natural disasters. The catastrophic loss of life and property has lead people to question why the United States is not better prepared for such disasters. In this Issue Brief, a leading expert on decision processes and low probability events discusses the reasons why people do not take protective measures voluntarily. In addition, he suggests ways to reduce losses through a disaster management plan that includes well-enforced building codes and land-use policies, insurance incentives, and long-term loans. PMID- 16708431 TI - Antibiotics, acne, and upper respiratory tract infections. AB - About two million people per year in the U.S. have acne severe enough to require treatment with antibiotics. Treatment frequently lasts for more than six months, prompting concerns about antibiotic resistance and other possible consequences of long-term antibiotic use, such as increased susceptibility to infections. This Issue Brief summarizes a large study that evaluates the risk for upper respiratory and urinary tract infections among adolescents and young adults treated with antibiotics for acne. PMID- 16708432 TI - Man with syncopal episodes and abnormal chest radiograph findings. Plombage therapy. PMID- 16708433 TI - The impact of maternal nutrition on the offspring. Proceedings of the 5th Nestle Pediatric Nutrition Workshop. April 2004. Beijing, Peoples Republic of China. PMID- 16708435 TI - Abstracts of the 151st Meeting of the Acoustical Society of America, Providence, Rhode Island, USA, 5-9 June 2006. PMID- 16708434 TI - Abstracts of Digestive Disease Week, May 20-24, 2006 and the ASGE annual postgraduate course, May 24-25, 2006, Los Angeles, California, USA. PMID- 16708436 TI - Abstracts of the Annual Meeting of the German Society for Cell Biology, March 29 April 1, 2006, Braunschweig, Germany. PMID- 16708437 TI - Reader questions benefit of nebulized opioids. PMID- 16708438 TI - One giant step back. PMID- 16708439 TI - Reader inquires about facilities that ban opioid medications. PMID- 16708440 TI - Proceedings of the Institute of Food Research symposium. June 28-July 1, 2005. Norwich, United Kingdom. PMID- 16708441 TI - Diagnosis and management of the metabolic syndrome. An American Heart Association/National Heart, Lung, and Blood Institute Scientific Statement. Executive summary. PMID- 16708442 TI - Medicaid: Medicaid: benefits and services--2005. End of Year Issue Brief. AB - Medicaid provides health care insurance for low-income children, some parents who meet income thresholds, pregnant women, the elderly and the disabled. In order to receive federal funds for Medicaid, each state must offer coverage for the following mandatory health care services: (1) Inpatient and outpatient hospital services; (2) Physician services; (3) Medical and surgical dental services; {4) Nursing facility services; (5) Home health care services; (6) Family planning services; (7) Rural health clinic services; (8) Laboratory and x-ray services; (9) Pediatric and family nurse practitioner services; (10) Federally qualified health center services; (11) Nurse-midwife services; (12) Early and periodic screening, diagnosis and treatment (EPSDT) services for individuals under age 21 In addition to the required services mentioned above, states can also choose to cover additional services under their Medicaid plans, and these often include prescription drugs, nonmedical or surgical dental services, clinic services, and vision and hearing services. It is up to each state to decide what to include with the mandated services to create their Medicaid benefit package. Because of the variety of people to whom state Medicaid programs offer health insurance, states have gone back and forth on what services to include in their benefit packages. Some states offer extensive benefits with few restrictions on access to services, while others offer a minimal number of optional benefits. Cost, most often, is the deciding factor on whether or not to offer coverage. PMID- 16708443 TI - [Severe refractory pulmonary hypertension after liver transplantation for hepatitis C liver cirrhosis]. PMID- 16708444 TI - [Report of the Working Group "Endoscopic Ultrasonography" on on 16.9.2005 in the framework of the of the 60th meeting of the German Society for Alimentary Metabolic Diseases 2005]. PMID- 16708445 TI - Health care providers and facilities: health facilities--2005. End of Year Issue Brief. AB - The mission of every hospital in America is to serve the health care needs of individuals in their communities, 24 hours a day, seven days a week. Their task, and the task of their medical staff, is to continually care for and to cure their patients. American health facilities are said to provide the best, most sophisticated, and most beneficial health care in the world. However, a hospital's ability to care for each patient who walks through their doors is continuously challenged on numerous fronts--the shortage of key hospital personnel, the increased cost of caring for the uninsured, the continued problem of medical errors, and the growth of niche and specialty hospitals. As of 2002, there were 5,794 registered hospitals in the United States, according to the most recent data available from the American Hospital Association (AHA). The AHA also states that there are 4,927 community hospitals, which includes nongovernmental, non-profit hospitals, investor-owned (for-profit) hospitals, and hospitals owned by state and local governments. The AHA defines community hospitals as all non federal, short-term general and other specialty hospitals. Specialty hospitals include obstetrics and gynecology, rehabilitation, orthopedic and other individually described specialty services. Statistics provided by the AHA indicate that the number of rural and urban community hospitals is approximately equal--2,178 rural hospitals compared to 2,749 urban hospitals. PMID- 16708446 TI - Shades of grey. PMID- 16708447 TI - Another incentive to lose weight. PMID- 16708448 TI - Ductal carcinoma in situ. PMID- 16708449 TI - Research in general practice. PMID- 16708450 TI - Health care providers and facilities: medical malpractice and tort reform--2005. End of Year Issue Brief. AB - As health care professionals continue to feel the crunch of rising malpractice insurance rates and increased jury awards, medical malpractice remains a priority for acute care professionals. Medical associations claim that rapidly increasing premiums and the declining number of insurers often lead physicians to stop practicing medicine or to relocate. This may lead to a shortage of physicians, particularly physicians who practice high-risk specialties such as neurology. The pressure to retain an adequate supply of health care professionals is particularly acute in rural areas. It is difficult to pinpoint the origins of the escalating cost of medical malpractice coverage. Insurers and physicians claim excessive litigation and overly generous jury awards have hardened the market. Trial lawyers and consumer advocacy groups assert insurance premium rates have not reflected increasing medical inflation or the payouts of jury awards during the last 30 years. The majority of states have some form of basic coverage requirement that medical malpractice insurers must offer. However, because of the complexities and variety of coverage plans, physicians often are unaware that gaps in coverage exist. As of May 2005, the American Medical Association (AMA) has declared a state PMID- 16708451 TI - The Brethren discuss genetic engineering. PMID- 16708452 TI - Theological reflections on genetic engineering. PMID- 16708453 TI - Long-term care: nursing home quality and safety--2005. End of Year Issue Brief. AB - In 2002, the Government Accountability Office reported that more than 1.7 million senior citizens resided in over 17,000 nursing homes across the United States. A 2003 Administration on Aging report predicted that number would increase dramatically as the "baby-boom" generation ages. Accordingly, legislators and nursing home administrators have striven to develop facilities that provide safe, high-quality eldercare to the nations' growing senior population. The Health Policy Tracking Service (HPTS) published a study in January--2005 Health Care Priorities Report--that depicts state lawmakers' concern for nursing home quality and safety. To policymakers, nursing home quality and safety is a very high priority, second only to Medicaid. The HPTS survey also indicated that 38 states planned to address senior facility safety in 2005 by adopting more stringent employee background checks, higher staffing standards and strict licensure requirements PMID- 16708454 TI - From meeting presentation to peer-review publication--a UK review. AB - INTRODUCTION: Annual academic surgical meetings provide a forum for the discussion of research. For the wide-spread dissemination of this information, peer-reviewed publication is required. The aim of this study was to compare the amount of presentations which go on to publication from 4 UK-based surgical meetings. MATERIALS AND METHODS: We determined whether a presentation had led to a successful publication using PubMed, a median of 28 months following each meeting. We compared the ASGBI publication rate with the meetings of the Vascular Surgical Society (VSSGBI), the Association of Coloproctology of Great Britain and Ireland (ACPGBI) and the British Transplantation Society (BTS). We also compared the median impact factor of journals used. RESULTS: The ASGBI and BTS had a similar rate of presentations resulting in publication, with 35% and 36% at 2 years, respectively. The VSS had a significantly greater proportion of presentations resulting in publication (54% at 2 years; P = 0.004), whilst the ACPGBI had significant fewer (24% at 2 years; P = 0.006). There was no difference in the median impact factors of the journals used between the meetings (Kruskal Wallis P = 0.883). CONCLUSIONS: There is a significant variation between meetings in terms of turning presentations into publications. However, the majority of abstracts have still not been fully published within 2 years of presentation at the meeting. PMID- 16708455 TI - Pharmaceuticals: pharmaceutical cost controls--2005. End of Year Issue Brief. AB - The enactment of the Omnibus Budget Reconciliation Act of 1990 (OBRA '90) gave states the option of offering pharmaceutical benefits within their Medicaid programs. But the law placed restrictions on states' flexibility to control what prescriptions they would cover and required the states to reimburse outpatient prescription drugs from manufacturers that signed rebate agreements with the U.S. Department of Health and Human Services. Forty-nine states--Arizona is excluded, based on its program structure--and the District of Columbia currently offer prescription drug coverage under the Medicaid Drug Rebate Program. During the past four years, states all over the country have been plagued with revenue shortfalls in their state Medicaid budgets. While the fiscal situation improved for most states in the 2004 legislative session, many states still face budget pressures in 2005. Compounding existing budget pressures are threats from the Bush Administration to shift increased costs of the Medicaid program on to the states. All things considered, the economic pressure of funding Medicaid is at the top of legislative agendas in 2005. As in previous years, states are attempting to reduce costs to their Medicaid programs by seeking savings in their pharmaceutical programs. Prescription drug costs are highly attributed as a contributing factor to the fiscal climate of state Medicaid programs. Currently, prescription drug spending outpaces that of every other category of health care and drug prices are rising faster than inflation. In response, states are instituting a variety of pharmaceutical cost control measures such as creating preferred drug lists (PDLs), negotiating supplemental rebates, forming bulk purchasing pools, promoting generic drug substitution and implementing price controls. As prescription drug cost containment tools have gained acceptance and momentum, they continue to be controversial. This issue brief explores the debate, history, methodology, utilization and 2005 legislative activity surrounding the most commonly used and emerging pharmaceutical cost control measures. PMID- 16708456 TI - Using a culturally relevant theory to recruit African American men for prostate cancer screening. PMID- 16708457 TI - Pharmaceuticals: drug importation--2005. End of Year Issue Brief. PMID- 16708458 TI - [Toxicity of orally ingested asbestos]. PMID- 16708459 TI - [Trans fatty acids]. PMID- 16708460 TI - Oral pathology quiz #50. Case number 4. Lateral periodontal cyst. PMID- 16708461 TI - Clinical update on root canal disinfection. AB - Endodontic disease is mediated by bacteria. Treatment goals should be directed to reducing the critical concentration of microbial irritants to the lowest level possible. Thorough canal instrumentation, with either stainless steel hand files or Ni-Ti rotary files, removes the bulk of tissue and microbial contamination, but adjunctive chemical agents are needed to optimize debridement. Current concepts support the following approaches: lubricating and chelating agents during cleaning and shaping; copious irrigation during all phases of instrumentation with 2.5% sodium hypochlorite, constantly refreshed; deep penetration of a side port, narrow gauge irrigating needle, constantly moving in and out of the canal space during each irrigation; removal of residual smear layer at completion of instrumentation prior to obturation. PMID- 16708462 TI - Trends in dentistry. PMID- 16708464 TI - Restorative treatment options for patients with severe orthodontic resorption. AB - The treatment of 2 patients with severe root resorption after orthodontic treatment is presented. Treatment options were reviewed. The multidisciplinary approach and treatment sequence that were used to achieve esthetic and functional results are discussed. PMID- 16708463 TI - Subepithelial connective tissue graft in a localized, plaque-induced ulcerative gingival recession: a case report. AB - The transposition of connective tissue to increase the zone of keratinized tissue or cover exposed root surfaces has become an integral part of the surgical dental practice. An effort to expand the surgical protocol to gingival recession of ulcerative etiology is presented. Parameters such as pathogenesis, clinical characteristic, and histopathology are analytically discussed. Furthermore, different aspects of the mucogingival therapy phase of the grafting procedure are presented. PMID- 16708466 TI - Esthetics vs evidence-based dentistry. AB - Esthetic procedures that require alteration of healthy dental tissue should be explained to the patient. Our professional obligation is to inform the patient of the risk-reward of a dental treatment that does not correct or enhance dental health. PMID- 16708465 TI - The benefits of glass ionomer self-adhesive materials in restorative dentistry. AB - Bonded, resin-based composite restorative materials have potential advantages. If the dentin bond achieved is not greater than the polymerization stress, loss of retention is likely, resulting in areas of microleakage and postoperative sensitivity. Class 5 lesions restored with no preparation have been used for testing the clinical performance of new adhesive restorative systems. Laboratory studies have demonstrated that bond strength varies according to the depth of dentin and the degree of calcification. Until the later generations of dentin bonding agents, retention rates for bonded, resin-based composites were typically more erratic and lower than glass ionomer (GI) and resin-modified glass ionomer (RMGI) restorative materials. Providing stress relief during setting is inherent in GI and RMGI materials, which helps to explain their good retention rates, despite their low bond strengths. GI and RMGI liners and lightly filled resin bonding agents provide similar stress relief. Current evidence supports the use of both RMGI and composite restorations placed with a liner of lightly filled resin in adhesive Class 5 restorations. Bonded, resin-based composite has the advantage of finishing to a high-gloss surface, making it more acceptable in areas of the mouth that are highly visible. PMID- 16708467 TI - A new bone surgery device: sinus grafting and periodontal surgery. AB - The PiezoSurgery device is a new instrument developed specifically for osseous surgery that has applications in various dental and medical surgical specialties. Using low-frequency piezoelectric ultrasonic vibration, the PiezoSurgery device precisely cuts bone without cutting soft tissue. This article presents 3 cases illustrating the use of the device in sinus grafting and periodontal osseous surgery. PMID- 16708469 TI - Ablation of cytoskeletal filaments and mitochondria in live cells using a femtosecond laser nanoscissor. AB - Analysis of cell regulation requires methods for perturbing molecular processes within living cells with spatial discrimination on the nanometer-scale. We present a technique for ablating molecular structures in living cells using low repetition rate, low-energy femtosecond laser pulses. By tightly focusing these pulses beneath the cell membrane, we ablate cellular material inside the cell through nonlinear processes. We selectively removed sub-micrometer regions of the cytoskeleton and individual mitochondria without altering neighboring structures or compromising cell viability. This nanoscissor technique enables non-invasive manipulation of the structural machinery of living cells with several-hundred nanometer resolution. Using this approach, we unequivocally demonstrate that mitochondria are structurally independent functional units, and do not form a continuous network as suggested by some past studies. PMID- 16708468 TI - Shear force at the cell-matrix interface: enhanced analysis for microfabricated post array detectors. AB - The interplay of mechanical forces between the extracellular environment and the cytoskeleton drives development, repair, and senescence in many tissues. Quantitative definition of these forces is a vital step in understanding cellular mechanosensing. Microfabricated post array detectors (mPADs) provide direct measurements of cell-generated forces during cell adhesion to extracellular matrix. A new approach to mPAD post labeling, volumetric imaging, and an analysis of post bending mechanics determined that cells apply shear forces and not point moments at the matrix interface. In addition, these forces could be accurately resolved from post deflections by using images of post tops and bases. Image analysis tools were then developed to increase the precision and throughput of post centroid location. These studies resulted in an improved method of force measurement with broad applicability and concise execution using a fully automated force analysis system. The new method measures cell-generated forces with less than 5% error and less than 90 seconds of computational time. Using this approach, we demonstrated direct and distinct relationships between cellular traction force and spread cell surface area for fibroblasts, endothelial cells, epithelial cells and smooth muscle cells. PMID- 16708470 TI - Growth, anisotropy, and residual stresses in arteries. AB - A simple phenomenological theory of tissue growth is used in order to demonstrate that volumetric growth combined with material anisotropy can lead to accumulation of residual stresses in arteries. The theory is applied to growth of a cylindrical blood vessel with the anisotropy moduli derived from experiments. It is shown that bending resultants are developed in the ring cross-section of the artery. These resultants may cause the ring opening or closing after cutting the artery in vitro as it is observed in experiments. It is emphasized that the mode of the arterial ring opening is affected by the parameters of anisotropy. PMID- 16708471 TI - The effect of longitudinal pre-stretch and radial constraint on the stress distribution in the vessel wall: a new hypothesis. AB - It is well known that blood vessels shorten axially when excised. This is due to the perivascular tethering constraint by side branches and the existence of pre stretch of blood vessels at the in situ state. Furthermore, vessels are radially constrained to various extents by the surrounding tissues at physiological loading. Our hypothesis is that the axial pre-stretch and radial constraint by the surrounding tissue homogenizes the stress and strain distributions in the vessel wall. A finite element analysis of porcine coronary artery and rabbit thoracic aorta based on measured material properties, geometry, residual strain and physiological loading is used to compute the intramural stresses and strains. We systematically examined the effect of pre-stretch and external radial constraint in both vessels. Our results show that both stretching in the axial direction and compression in the radial direction lead to a more homogeneous strain and stress state in the blood vessel wall. A "uniform biaxial strain" hypothesis is proposed for the blood vessel wall and the ramifications are discussed. PMID- 16708472 TI - Catch bonds: physical models and biological functions. AB - Force can shorten the lifetimes of receptor-ligand bonds by accelerating their dissociation. Perhaps paradoxical at first glance, bond lifetimes can also be prolonged by force. This counterintuitive behavior was named catch bonds, which is in contrast to the ordinary slip bonds that describe the intuitive behavior of lifetimes being shortened by force. Fifteen years after their theoretical proposal, catch bonds have finally been observed. In this article we review recently published data that have demonstrated catch bonds in the selectin system and suggested catch bonds in other systems, the theoretical models for their explanations, and their function as a mechanism for flow-enhanced adhesion. PMID- 16708474 TI - Evaluation of tension in actin bundle of endothelial cells based on preexisting strain and tensile properties measurements. AB - Actin bundles in vascular endothelial cells (ECs) play a critical role in transmitting intracellular forces between separate focal adhesion sites. However, quantitative descriptions of tension level in single actin bundles in a physiological condition are still poorly studied. Here, we evaluated magnitude of preexisting tension in a single actin bundle of ECs on the basis of measurements of its preexisting stretching strain and tensile properties. Cultured ECs expressing fluorescently-labeled actin were treated with detergents to extract acin bundles. One end of an actin bundle was then dislodged from the substrate by using a microneedle, resulting in a shortening of the actin bundle due to a release of preexisting tension. Assuming the shortened actin bundle reached its non-stress state, preexisting stretching strain was determined to be 0.24 on average. A tensile test of the dislodged single acin bundle was conducted with a pair of cantilevers to measure the force required for stretching it up to the original length, yielding an estimate of preexisting tension in the actin bundle. The magnitude of the preexisting tension, 4 nN on average, was comparable to previously reported data of the traction force generated by adherent cells at single adhesion sites to keep cell integrity. The Young's modulus of the isolated actin bundle was estimated to be approximately 300 kPa from the tensile tests together with evaluation of average diameter of the isolated actin bundle based on transmission electron microscopy. These data will contribute to better understanding of intracellular stress transmission mechanism in ECs. PMID- 16708473 TI - Biophysical approaches for studying the integrity and function of tight junctions. AB - Cell-cell adhesion is an extremely important phenomenon as it influences several biologically important processes such as inflammation, cell migration, proliferation, differentiation and even cancer metastasis. Furthermore, proteins involved in cell-cell adhesion are also important from the perspective of facilitating better drug delivery across epithelia. The adhesion forces imparted by proteins involved in cell-cell adhesion have been the focus of research for sometime. However, with the advent of nanotechnological techniques such as the atomic force microscopy (AFM), we can now quantitatively probe these adhesion forces not only at the cellular but also molecular level. Here, we review the structure and function of tight junction proteins, highlighting some mechanistic studies performed to quantify the adhesion occurring between these proteins and where possible their association with human diseases. In particular, we will highlight two important experimental techniques, namely the micropipette step pressure technique and the AFM that allow us to quantify these adhesion forces at both the cellular and molecular levels, respectively. PMID- 16708475 TI - Control of the direction of lamellipodia extension through changes in the balance between Rac and Rho activities. AB - The direction in which cells extend new motile processes, such as lamellipodia and filopodia, can be controlled by altering the geometry of extracellular matrix adhesive islands on which individual cells are cultured, thereby altering mechanical interactions between cells and the adhesive substrate [Parker (2002)]. Here we specifically investigate the intracellular molecular signals that mediate the mechanism by which cells selectively extend these processes from the corners of polygonal-shaped adhesive islands. Constitutive activation of the small GTPase Rac within cells cultured on square-shaped islands of fibronectin resulted in the elimination of preferential extension from corners. This loss of directionality was accompanied by a re-distribution of focal adhesions: the large focal adhesions normally found within the corner regions of square cells were lost and replaced by many smaller focal contacts that were distributed along the entire cell perimeter. Inhibition of the small GTPase, Rho, using C3 exoenzyme blocked lamellipodia extension entirely. However, inhibition of Rho signaling in combination with ectopic Rac activation rescued the corner localization of motile processes and focal adhesions. These results suggest that the ability of cells to sense their physical surroundings and respond by moving in a spatially oriented manner is mediated by a balance between Rho and Rac activities. PMID- 16708477 TI - Don't rob the cradle. PMID- 16708476 TI - Melanoma cell extravasation under flow conditions is modulated by leukocytes and endogenously produced interleukin 8. AB - Attachment of tumor cells to the endothelium (EC) under flow conditions is critical for the migration of tumor cells out of the vascular system to establish metastases. Innate immune system processes can potentially promote tumor progression through inflammation dependant mechanisms. White blood cells, neutrophils (PMN) in particular, are being studied to better understand how the host immune system affects cancer cell adhesion and subsequent migration and metastasis. Melanoma cell interaction with the EC is distinct from PMN-EC adhesion in the circulation. We found PMN increased melanoma cell extravasation, which involved initial PMN tethering on the EC, subsequent PMN capture of melanoma cells and maintaining close proximity to the EC. LFA-1 (CD11a/CD18 integrin) influenced the capture phase of PMN binding to both melanoma cells and the endothelium, while Mac-1 (CD11b/CD18 integrin) affected prolonged PMN melanoma aggregation. Blocking E-selectin or ICAM-1 (intercellular adhesion molecule) on the endothelium or ICAM-1 on the melanoma surface reduced PMN facilitated melanoma extravasation. Results indicated a novel finding that PMN facilitated melanoma cell arrest on the EC could be modulated by endogenously produced interleukin-8 (IL-8). Functional blocking of the IL-8 receptors (CXCR1 and CXCR2) on PMN, or neutralizing soluble IL-8 in cell suspensions, significantly decreased the level of Mac-1 up-regulation on PMN while communicating with melanoma cells and reduced melanoma extravasation. These results provide new evidence for the complex role of hemodynamic forces, secreted chemokines, and PMN-melanoma adhesion in the recruitment of metastatic cancer cells to the endothelium in the microcirculation, which are significant in fostering new approaches to cancer treatment through anti-inflammatory therapeutics. PMID- 16708478 TI - Legislating integrity. PMID- 16708479 TI - Mixing it up. Harmless levels of chemicals prove toxic together. PMID- 16708481 TI - Pumping coal. PMID- 16708480 TI - Chaos in the crater. PMID- 16708482 TI - Enhanced armor. PMID- 16708485 TI - Android science. PMID- 16708483 TI - Light work. Better solar nanotubes to split water for hydrogen. PMID- 16708484 TI - SHAM scam. PMID- 16708486 TI - The first few microseconds. PMID- 16708487 TI - Tapping the computing power of biological molecules gives rise to tiny machines that can speak directly to living cells. PMID- 16708488 TI - The birth of the mighty Amazon. PMID- 16708490 TI - Giant telescopes of the future. PMID- 16708489 TI - Blockbuster dreams. PMID- 16708491 TI - Shutting down Alzheimer's. PMID- 16708492 TI - When slide rules ruled. PMID- 16708493 TI - Robot mowers. Cutting work. PMID- 16708494 TI - What makes a revolution? PMID- 16708495 TI - How do salt and sugar prevent microbial spoilage? PMID- 16708496 TI - Disparities between white and African-American children in immunization coverage. AB - INTRODUCTION: A recent study has shown that the national-scale difference in immunization coverage between non-Hispanic white (abbreviated "white") and non Hispanic African-American (abbreviated "African-American") children aged 19-35 months in the United States has increased by about 1 percentage point annually. We examined how this widening gap differs with geography and income. METHODS: We used data from the National Immunization Survey, 1998-2003, a national telephone survey. We examined differences between white and African-American children in immunization coverage within income groups (at or above versus below the federal poverty level) for each census region (northeast, south, midwest and west). We tested the hypothesis of constant disparity over time. RESULTS: Among households at or above the federal poverty level in the northeast census region, disparity is widening (white coverage minus African-American coverage was -0.5 in 1998 but 15.5 in 2003). Among household at or above the federal poverty level in the midwest census region, disparity is narrowing (white coverage minus African American coverage was 13.9 in 1998 but 2.5 in 2003). We found no significant evidence of a trend in other groups. CONCLUSIONS: Widening national-level disparity in immunization coverage is primarily attributable to trends in the northeast census region. Addressing the widening disparity in coverage requires new strategies that consider current social and economic contexts. PMID- 16708497 TI - The relationship between asthma and overweight in urban minority children. AB - BACKGROUND: This study was performed to determine the relationship between overweight [body mass index (BMI) > or = 85th percentile] and asthma as determined by spirometry. METHOD: Spirometry was performed according to the American Thoracic Society guidelines, and BMI was calculated. Asthma was defined as a forced expiratory volume in 1 second (FEV1) <80% predicted and FEV1/forced vital capacity (FVC) >5% lower than predicted for age and sex. RESULTS: One hundred-nine children (age 14.7 +/- 1.6 years) were enrolled. Eighty children (73%) were African-American, and 29 children (27%) were white. Fifty-eight (53%) children were overweight. Twelve (11%) children, of whom nine (75%) were overweight, met the criteria for asthma. Baseline FEV1 percent predicted (87 +/- 6% vs. 83 +/- 7%, p=0.03), FEV1/FVC (93 +/- 6 vs. 87 +/- 8, p<0.001), and FEV1 percent predicted following albuterol administration (94 +/- 7 vs. 89 +/- 7%, p=0.03) were all lower in overweight children. Children with asthma were almost 1.5 times more likely to be overweight compared with children without asthma (relative risk: 1.49, 95% confidence interval: 1.015-2.17). CONCLUSIONS: Inner city children are more likely to be overweight compared to the general population. Asthma is a risk factor for overweight in these children. PMID- 16708498 TI - Immunomodulation therapy in children with chronic hepatitis B. AB - PURPOSE: The aim of this study is to investigate the effects of HBsAg vaccine and levamisole on virological indicators in naive patients suffering from chronic hepatitis B (CHB) and in healthy carriers of hepatitis B. METHOD: Vaccination and treatment with levamisole were applied to 93 minor patients in total, 43 of them inactive CHB carriers and 50 patients suffering from CHB. RESULTS: 15 (30%) of 50 patients who had high ALT values in the beginning of the study had normal values after treatment. In nine (12%) patients, posttreatment ALT values were higher than pretreatment values, and six (10%) patients showed HBV-DNA loss. In spite of the presence of 50 (54%) HBeAg-positive patients before treatment, 17 (34%) patients proved to be HBeAg-negative after treatment. HBeAg sero-conversion was seen in 10 (20%) cases. In two (2%) patients, HBsAg sero-conversion occurred. CONCLUSION: It was found that treatment with levamisole and vaccine had positive effects on CHB patients and healthy carriers with respect to HBV DNA loss, HBeAg sero-conversion and ALT normalization. The viral load increases and ALT increases that occurred in certain cases were thought to be related to the early immune response. It was determined that combined levamisole and vaccine therapy had no additional positive effect. PMID- 16708499 TI - Parental perspectives on influenza vaccination of children with chronic medical conditions. AB - BACKGROUND: Minorities and those living in the inner city have a higher respiratory disease burden than other groups. Yet, influenza vaccination rates among all children with chronic medical conditions remain low. METHODS: Parents of 2-13-year-old children with high-risk medical conditions from health centers in low-income urban neighborhoods completed a mailed survey. Immunization status from medical records was used to calculate validity measures. Survey data are presented for those whose vaccination status was concordant between parental report and the medical record (n=183). RESULTS: Parent-reported influenza vaccination versus medical record review showed 84.9% sensitivity, 68.7% specificity, 49.1% positive predictive value and 92.7% negative predictive value, with a kappa of 0.43. Vaccination rate was 30.6%. Medical record-verified influenza vaccination was associated with parental beliefs that the doctor recommends a flu shot (OR, 40.9; 95% Cl, 9.0-184.9) and that relatives recommend a flu shot (OR, 4.3; 95% Cl, 1.7-10.5), and was less likely if the parent believed that the child will get the flu if a household member is infected (OR, 0.2; 95% Cl, 0.1-0.6). CONCLUSIONS: The message that influenza vaccination is important to protect children with chronic medical conditions may be relayed through physician recommendation or a relative's suggestion and may be more effective if it addresses vaccine efficacy issues. PMID- 16708500 TI - Racial differences in pediatric renal transplantation-24-year single center experience. AB - Racial disparities in quality of health are major issues affecting healthcare delivery. While substantial studies have been done in adults with end-stage renal disease (ESRD), similar studies are lacking in pediatric patients with ESRD. We retrospectively analyzed our transplant database from 1978-2002 to identify racial trends in a pediatric ESRD population. Significant racial differences were noted in number of pre-emptive transplants, type of graft received, height at transplant and causes of ESRD. PMID- 16708501 TI - Child and adolescent violent deaths: an epidemiologic investigation. AB - OBJECTIVES: An apparent increase in violent deaths among children and adolescents in Detroit, MI in 2002 prompted a coordinated epidemiologic investigation involving federal, state and local organizations. METHODS: A descriptive analysis of cases and violent deaths (homicide, suicide or firearm-related) among juveniles <17 years was conducted, along with a case-control study using records from the medical examiner, police, schools and social service agencies. RESULTS: Twenty-nine cases were identified. Median age was 10 years (range 1 day-16 years), and 15 (52%) were male. There were 25 homicides, two suicides and two unintentional firearm-related deaths. Nine (31%) homicides resulted from child abuse and neglect, and eight (28%) were among bystanders. The most common mechanism of fatal injury was firearm (63%). Victims' families were more likely to have a history of familial violence, prior contact with the state social services agency, > or = 2 adults and > or = 4 persons in the household (P<0.05). CONCLUSIONS: The 2002 deaths did not represent a statistically significant increase from previous years. Several findings were remarkable: the proportion of deaths among bystanders, females and children age <5. PMID- 16708502 TI - Prenatal ultrasound: a tale of two cities. AB - At the beginning of the 21st century, community-based physicians have a technology which is safe, noninvasive and relatively inexpensive. It allows immediate access to visual data inside the body for the evaluation of health and disease. Prenatal ultrasound is vital for the efficient provision of preventive care even in community health centers, where physicians may not be the delivering physicians. To assess the frequency of this diagnostic technology, a longitudinal study of 36 months tabulated prenatal ultrasound scans performed in two community based offices providing generalist healthcare from 2001 through 2003. Between the two offices, 68,938 patients were seen, 679 deliveries were attended and 1,286 medically indicated prenatal ultrasound examinations were performed. Prenatal ultrasound scans--compared to other common office procedures, such as electrocardiograms, chest radiographs, circumcisions, blood sugar checks, Papanicolaous and colposcopies--emerged as one of the most frequently preformed diagnostic tests in the office. Based on these results, medical educators preparing generalist physicians for community medicine may want to consider some training in prenatal ultrasound. Future research can confirm the generalizability of the findings of this pilot study in two urban health centers. PMID- 16708503 TI - Residential racial concentration and birth outcomes by nativity: do neighbors matter? AB - OBJECTIVES: We examined the associations of residential segregation with poor birth outcomes (low birthweight, preterm) and with perinatal risk markers (maternal age, education and marital status, prenatal care and substance use, presence of paternal information on birth certificate) for foreign- and native born black women in the Minnesota seven-county metropolitan area. METHODS: Data were from 1990-1999 Minnesota birth certificates linked to the 1990 U.S. census. We used multivariable logistic regression to examine the association of perinatal risk markers, low birthweight and preterm birth for foreign- and native-born black women by residential black concentration. RESULTS: Native-born black women had a higher prevalence of risk markers and were at almost 1.5 times the risk of foreign-born blacks for delivery of low-birthweight or preterm infants. Risk markers and poor birth outcomes were most prevalent in medium and high-black concentration areas than low-concentration areas. Preterm birth was slightly positively associated with residential black concentration. CONCLUSIONS: Native born black women were at higher risk than foreign-born women for delivery of preterm and low-birthweight infants. Residential black concentration was associated with risk markers and only slightly associated with preterm birth. Further study of why birth outcome differentials exist by nativity and residential black concentration may identify opportunities for community-based public health interventions. PMID- 16708504 TI - Arterial stiffness variations by gender in African-American and Caucasian children. AB - OBJECTIVE: Most arterial stiffness studies have been conducted in adult populations as a part of the aging process in the arterial system. Arterial stiffness is an important early marker of disease identification that may lead to improved cardiovascular health. The aim of this study was to assess the gender and ethnic differences in the arterial stiffness levels among children and adolescents. DESIGN: From a subgroup of schoolchildren who participated in a prospective cohort study in Minnesota, Caucasian and African-American children who completed 16 timed visits were included in this report (n=487). The participants were followed from 1978 (7.68 +/- 0.72 years) to 1987 (16.65 +/- 0.71 years). A surrogate measure of arterial stiffness-arterial pulse pressure (APP in mmHg)--was used. RESULTS: Adjusted APP differences started to appear around 12.67 years and persisted throughout the study. Boys consistently had higher APP levels than the girls. Ethnic differences in adjusted APP levels were observed at an earlier age (7.68 years) but did not persist after age 10. CONCLUSION: APP levels were different between gender and ethnic groups in youth. These early indications of arterial stiffness warrant further exploration of arterial stiffness etiology. PMID- 16708505 TI - The pediatric resident training on tobacco project: interim findings. AB - OBJECTIVES: The Pediatric Residency Training on Tobacco Project is a four-year randomized prospective study of the efficacy of training pediatric residents to intervene on tobacco. At the start of the study (baseline), the pediatric residents uniformly agreed that environmental tobacco smoke (ETS) and tobacco use pose serious threats to the health of young people, and pediatricians should play a leadership role in the antismoking arena. However, very few went beyond advising patients and parents to modify their behavior by providing actual assistance, and many of them lacked necessary tobacco intervention skills and knowledge. We hypothesized that both standard training and special training programs would yield positive changes in intervention skills and activities, although the changes would be greater in residents exposed to the special training condition. In the present report, we present two-year outcome data from the resident tobacco surveys and objective structured clinical examinations (OSCEs) administered to independent waves of third-year residents in each experimental condition at baseline and year 2. METHODS: Fifteen pediatric residency training programs in the New York/New Jersey metropolitan area were assigned randomly to special and standard training conditions (eight to special and seven to standard training). Resident tobacco surveys and OSCEs were administered to third-year residents at the start of the training programs (baseline) and at years 1 and 2 of the study. Comparisons between sequential waves of third-year residents with no (baseline) or two-year exposure to the training programs permitted assessment of changes in resident beliefs, intervention activities and intervention skills within each experimental condition. RESULTS: By year 2, the residents associated with each training condition benefited from the training program, but the annual surveys and OSCEs revealed more significant positive changes for waves of residents in the special training condition. Most important, third-year residents exposed to the special training condition for two years were more likely than comparable residents in the standard training condition to reveal significant increases in the degree to which they provided active assistance for modifying smoking and ETS. CONCLUSIONS: The two-year findings from the pediatric tobacco project are encouraging and suggest that the special training program is efficacious, although aspects of the program in need of improvement were identified. PMID- 16708506 TI - Auditory functions in children at schools for the deaf. AB - OBJECTIVE: To evaluate auditory functions in children at schools for the deaf in Turkey. DESIGN: A total of 218 children who were attending the school for deaf children were involved in the study. Familial and medical histories were obtained, and otoscopic examinations were performed. Immittance audiometry, acoustic reflex testing, pure tone audiometry, otoacoustic emission and auditory brain stem response tests were performed. RESULTS: The mean age of identification of hearing loss was 48 months. Impacted wax was the most common otoscopic finding that was seen in 49 (22.47%) of children. Nontype-A tympanograms were found in 18 (8.25%) of children. One-hundred-eighty-nine (86.69%) children had profound hearing loss, and 29 (10.3%) had severe hearing loss on pure tone audiometry. On auditory brain stem response testing, 192 (88.07%) children had profound hearing loss, and 26 (11.41%) had severe hearing loss. Only one child had auditory neuropathy/dys-synchrony, as his otoacoustic emission results were normal without synchronous auditory brain stem responses. The hearing threshold levels were found >105 dB in 28 children only with pure tone audiometry. CONCLUSION: Early auditory screening is necessary to identify the children at risk. All hearing disorders cannot be detected by subjective or objective audiometric tests only. Pure tone audiometry still has a role in determining hearing threshold levels. The audiological research directions should be directed towards routine pure tone audiometry, otoacoustic emission and auditory brain stem response assessment for all hearing impaired children to enable an successful treatment. PMID- 16708507 TI - Meningitis caused by Salmonella panama in infants. AB - Salmonella panama is group-D nontyphi salmonella strongly associated with invasive infection, including meningitis. So far, no case of S. panama meningitis has been reported from the United States, and none has ever been reported in babies >3.5 months of age. To the best of our knowledge, we are reporting the first such case in English-language literature. PMID- 16708508 TI - Natal teeth: a review. AB - The incidence of natal teeth is approximately 1:2,000 to 1:3,000 live births. The most commonly affected teeth are the lower primary central incisors. Natal teeth usually occur in pairs. The eruption of more than two natal teeth is rare. The majority of natal teeth represent the early eruption of normal primary deciduous dentition. Less than 10% of natal teeth are supernumerary. Natal teeth might resemble normal primary dentition in size and shape; however, the teeth are often smaller, conical and yellowish, and have hypoplastic enamel and dentin with poor or absent root formation. Complications include discomfort during suckling, sublingual ulceration, laceration of the mother's breasts and aspiration of the teeth. A dental roentgenogram is indicated to differentiate the premature eruption of a primary tooth from a supernumerary tooth. Tooth extraction is indicated if the tooth is supernumerary or excessively mobile. If the tooth does not interfere with breastfeeding and is otherwise asymptomatic, no treatment is necessary. PMID- 16708509 TI - Deficits in diagnosis, treatment and continuity of care in African-American children and adolescents with ADHD. AB - Despite the evidence that attention-deficit/hyperactivity disorder (ADHD) is not just a diagnosis of whites, it often goes undiagnosed and is underresearched in the African-American population. There are higher rates of delinquency, incarceration, teen pregnancy and sexually transmitted diseases associated with inadequate or delayed treatment of ADHD. Afrcan Americans generally respond well to treatments, but access to evaluation, medication and psychotherapy is limited or absent for many, The purpose of this research is to compare descriptive characteristics of African-American children with ADHD to age-matched Caucasian children with the same diagnosis. Age at diagnosis, treatment offered, perception of outcome, adherence, comorbid symptoms and frequency of follow-up were collected retrospectively from charts of children treated in the sections of child and adolescent psychiatry and pediatric neurology. PMID- 16708510 TI - Race disparities in childhood asthma: does where you live matter? AB - OBJECTIVE: This study investigates whether racial/ethnic disparities in childhood asthma prevalence can be explained by differences in family and neighborhood socioeconomic position (SEP). METHODS: Data were from the 2001 Rhode Island Health Interview Survey (RI HIS), a statewide representative sample of 2,600 Rhode Island households, and the 2000 U.S. Census. A series of weighted multivariate models were fitted using generalized estimating equations (GEE) for the logistic case to analyze the independent and joint effects of race/ethnicity and SEP on doctor-diagnosed asthma among 1,769 white, black and Hispanic children <18 years old. RESULTS: Compared with white children, black children were at increased odds for asthma and this effect persisted when measures of family and neighborhood SEP were included in multivariate models (AOR: 2.49; 95% Cl: 1.30 4.77). Black children living in poverty neighborhoods had substantially higher odds of asthma than Hispanic and white children in poverty areas and children in moderate- and high-income neighborhoods (AOR: 3.20: 95% Cl: 1.62-6.29). CONCLUSION: The high prevalence of asthma among black children in poor neighborhoods is consistent with previous research on higher-than-average prevalence of childhood asthma in poor urban minority communities. Changing neighborhood social structures that contribute to racial disparities in asthma prevalence remains a challenge. PMID- 16708511 TI - Does a multifaceted environmental intervention alter the impact of asthma on inner-city children? AB - OBJECTIVE: To evaluate the impact of a multifaceted environmental and educational intervention on the indoor environment and health in 5-12-year-old children with asthma living in urban environments. DESIGN: Changes in indoor allergen levels and asthma severity measurements were compared between children who were randomized to intervention and delayed intervention groups in a 14-month prospective field trial. Intervention group households received dust mite covers, a professional house cleaning, and had roach bait and trays placed in their houses. RESULTS: Of 981 eligible children, 410 (42%) were enrolled; 161 (40%) completed baseline activities and were randomized: 84 to intervention and 77 to delayed intervention groups. At the study's end, dust mite levels were 163% higher than at baseline for the delayed intervention group. Overall asthma severity scores did not change. However, the median functional severity score (FSS) component of the severity score improved more in the intervention group (33% vs. 20%) than in the delayed intervention group. At the study's end, the median FSSs for the intervention group improved 25% compared with the delayed intervention group, (p<0.01). Differences between groups for medication use, emergency department (ED) visits or hospitalization were not significant. CONCLUSIONS: Despite low retention, the intervention resulted in decreased dust mite allergen levels and increased FSSs among the intervention group. The interventions probably contributed to the improvements, especially among the more severely affected children. This study highlights the complexities of designing and assessing the outcomes from a multifaceted asthma intervention. PMID- 16708512 TI - Effects of topical emollient therapy on infants at or less than 27 weeks' gestation. AB - OBJECTIVE: To evaluate the effects of topical emollient therapy on fluid intake, urine output, serum electrolytes, glucose, bilirubin and other outcome measures of neonates < or = 27 weeks' gestational age (GA) with birthweight (BW) <1,000 g. METHODS: We reviewed medical records of 18 infants treated with topical emollient Aquaphor, and 36 BW- and GA-matched control infants that were not treated with similar topical emollient. RESULTS: Characteristics of the study and control infants were similar: BW: 698 +/- 144 g vs. 732 +/- 134 g, GA: 25.5 +/- 1.33 weeks vs. 25 +/- 1.6 weeks and Score for Neonatal Acute Physiology (SNAP) 14.3 +/ 5.1 vs. 14.6 +/- 7.8, respectively. Fluid intake was lower and urine output was significantly better in Aquaphor-treated infants during the first two weeks of life. Peak serum potassium and bilirubin values were also lower in the study infants. Insulin use, patent ductus arteriosus (PDA), inltraventricular hemorrhage (IVH), retinopathy of prematurity (ROP), sepsis and duration of ventilator/oxygen use weresimilar among the groups. CONCLUSION: Infants < or = 27 weeks' gestation who had Aquaphor applied to their skin from birth required less fluids and had better urine output. These infants had lower serum potassium and bilirubin values during their first two weeks of life. Therefore, we conclude that topical Aquaphor application to thee skin is beneficial for fluid and electrolyte balance in extreme preterm infants. PMID- 16708513 TI - Eliminating child health disparities: a call to action. PMID- 16708514 TI - Public health and Hurricane Katrina: lessons learned and what we can do now. AB - Keith C. Ferdinand, MD, FACC, presently resides in Atlanta, GA, displaced from his native New Orleans by Hurricane Katrina, September 2005. The hurricane destroyed hiss cardiovascular center and severely damaged his home. In Atlanta, he is director of the Association of Black Cardiologists Hurricane Katrina Relief. PMID- 16708515 TI - Sexually transmitted disease prevention in adolescents and young adults. AB - OBJECTIVE: Sexuality is one of the most pervasive aspects of the human life cycle. It warrants attention in childhood, adolescence and adulthood, and is an integral part of each health maintenance visit. Given this, it is unfortunate that U.S. medical schools do not offer more training in soliciting sexual histories and initiating dialogue about sexually healthy lifestyles. METHODS: Fourth-year medical student completed an adolescent and young-adult sexually transmitted disease elective. RESULTS: This elective allows medical students to confront personal biases and discomfort levels yet remain objective. It is also an opportunity to explore how the provider's body language and facial expressions can discourage information disclosure. Although sexuality is very prevalent in our society today, patients may still be apprehensive about discussing details of their sexual practices. Therefore, it becomes incumbent upon the physician to create an environment free from personal prejudice in order to best serve the patient. CONCLUSIONS: In order to promote sexual health awareness in a society that can be conservative and judgmental in this subject matter, it is essential to train all healthcare providers to lead discussions, educate patients and provide treatment in hopes that sexual health promotion will become as important as other socially accepted healthcare concerns. PMID- 16708516 TI - Danger of multiple magnets beyond the stomach in children. AB - Examples from our recent experience, as well as several reports in surgical and pediatric journals, proclaim the danger in children whenever more than one swallowed magnet travels beyond the stomach. They may attract across the thin walls of small bowel, causing ischemia, necrosis and perforation into the peritoneum. The radiologist or other healthcare worker seeing magnets in the abdomen on radiographs should consider the situation an emergency and recommend surgical treatment as soon as possible. PMID- 16708518 TI - Urethral duplication with a perineal opening in a four-year-old boy. AB - Urethral duplication (UD) is a rare congenital anomaly. In UD, there is in addition to a normally positioned urethra an accessory urethra arising from the bladder, bladder neck or the proximal urethra and terminating anywhere between the base of the penis along the ventral surface and the glans penis. The embryologic development of UD remains to be clearly elucidated and the etiology of the condition is unknown. We present our experience with a 4 year old boy who presented with urethral duplication. PMID- 16708517 TI - An unexpected finding in an eight-year-old child with cerebral palsy and weight loss. AB - Poor weight gain is frequently seen in children with cerebral palsy. This is most commonly due to poor oral-motor coordination, resulting in inadequate caloric intake. We present the case of an eight-year-old girl with Aicardi syndrome who had been an exclusively oral feeder and who was noted to have had a 10-lb weight loss over the previous months when she was admitted to the hospital with seizures. Due to ongoing diminished oral intake, a barium swallow was performed, which revealed a filling defect. The mass that was removed at surgery proved to be a lactobezoar, a very uncommon finding in a child of this age. PMID- 16708519 TI - Sudden deterioration in the renal function of an African child with cyanotic congenital heart disease. AB - Nephrotic syndrome (NS) and deteriorating renal function are associated with congenital cyanotic heart disease (CCHD). We describe a nine-year-old African male child with CCHD, NS and deteriorating renal function. He presented with two weeks' history of progressive generalized body swelling, oliguria and orthopnea. Oliguria did not improve, and the generalized body swelling did not subside with the administration of diuretics. Dipstick urinalysis remained 3+. He was referred to a facility that offers renal replacement therapy in view of worsening renal status. This is the first description known to us of deteriorating renal function in an African and in a child with CCHD, implying that the complication can develop in children or commences in childhood. PMID- 16708520 TI - Analysis of shrinkage of different gutta-percha types using optical measurement methods. AB - The aim of this study was to compare the shrinkage of alpha- and beta-gutta percha of the Multifill-system with commercial gutta-percha. Ten gutta-percha blocks of each of the three types were used. Speckle pattern shearing interferometry was used to prove that no trapped air and material defects were present in the specimens. The optical triangulation method was applied to assess shrinkage. The three gutta-percha types were examined after heating up to 90 degrees C and cooling down to 35 degrees C. Commercial gutta-percha showed less shrinkage (6.5%) than alpha- (7.2%) and beta-gutta-percha (7.3%; p=0.0051 for both comparisons; Wilcoxon's signed rank test). There was a significant difference between alpha- and beta-gutta-percha (p=0.0093; Wilcoxon's signed rank test). In the range of 45 degrees C to 40 degrees C, alpha- and beta-gutta-percha revealed the highest shrinkage (2.2%/5 degrees C and 2.1%/5 degrees C, respectively). Commercial gutta-percha showed the highest shrinkage (1.1%/5 degrees C) in the range of 55 degrees C to 40 degrees C. Overall, shrinkage of all types of gutta-percha was higher than could be assumed from former studies. PMID- 16708522 TI - Production of fluoridated salt. AB - Sodium and potassium fluoride are used for the fluoridation of household salt. Addition of fluoride is carried out either by the wet or the dry process. Qualitatively good fluoridated salt can be produced by using either method. In the wet process, a solution of potassium fluoride is mixed homogeneously with the salt. For the dry mixture, only sodium fluoride with a small granule size is suitable. Apart from the mixing methods used by the larger salt manufacturers, there are low-cost solutions for very small producers. PMID- 16708521 TI - Fluoride excretion of adults living in border regions with either water or salt fluoridation. AB - The canton of Basel-Stadt was the only canton in Switzerland which introduced drinking water fluoridation (DWF) at 1 ppm (mg/l). All other cantons have relied on fluoridated domestic salt at 250 ppm F as the main vehicle for basic fluoride exposure. It has been suggested that persons living and working in the DWF areas or persons commuting to the DWF areas may be exposed to higher than optimal doses of fluoride. The objective of this present study was to determine the urinary fluoride excretion of adults living and or working in neighboring areas of either salt or water fluoridation. In this study, 24-hour urine was collected from 69 healthy subjects and tested for fluoride concentration. The mean fluoride concentration for all participants was 0.55 +/- 0.25 ppm (mg/l) ranging from 1.14 to 0.09 ppm. The mean fluoride excretion was 0.95 +/- 0.47 mg F/d ranging from 0.18 to 2.12 mg F/d. The 33 subjects living in a DWF region showed a mean urine fluoride concentration of 0.64 +/- 0.24 ppm (mg/l) and a mean fluoride excretion of 1.14 +/- 0.48 mg F/d. Those 36 subjects living in a region without DWF showed a mean urine fluoride concentration of 0.47 +/- 0.24 ppm (mg/l) and a mean fluoride excretion of 0.78 +/- 0.40 mg F/d. A significant difference between the two means of the groups living in regions with or without DWF was detected when the Mann-Whitney statistical test was applied (p < 0.005). The combined intake of fluoridated drinking water and fluoridated table salt in the sub-group of 11 subjects who commuted showed an overall increase in fluoride urine concentration. The measured values, however, were not significantly different from the other sub groups. PMID- 16708523 TI - Legal aspects of fluoride in salt, particularly within the EU. AB - In seven European countries there are national legal regulations, or salt producers have obtained individual authorisations, for the production and marketing of fluoridated edible salt. On the basis of EU mutual recognition rules, there are other countries which import fluoridated edible salt. All European countries practise salt fluoridation on a voluntary basis. In the near future, a European regulation is expected to supersede the national conditions. PMID- 16708524 TI - [The "globulomaxillary cyst" a specific entity or a myth?]. AB - The following review investigates the term and concept of the globulomaxillary cyst as a correct clinico-pathological diagnosis to describe a so-called fissural cyst said to be caused by epithelial entrapment between the nasal and maxillary process. After analyzing the available literature it has to be concluded that neither from an embryologic nor from a clinical or pathohistological standpoint the term globulomaxillary cyst represents a real entity by itself. Therefore, globulomaxillary cysts have to be diagnosed alternatively after a thorough clinical, radiological and histological examination as other odontogenic cysts like dentigerous cysts or odontogenic keratocysts, odontogenic tumors like ameloblastoma, central giant cell tumors, solitary bone cysts, etc. PMID- 16708525 TI - [Vertical and horizontal jaw relations in reconstructive dentistry. A critical review]. AB - Despite comprising one of the routine duties of dental practitioners, the methods used for the determination of the vertical and horizontal jaw position differ considerably from dentist to dentist. For the overwhelming majority of patients, however, the techniques generally applied for the maxillomandibular relationship record appear to lead to acceptable results. No method seems to be clearly superior to the other. As far as the determination of the horizontal mandibular position (centric relation interocclusal record) is concerned, bimanual manipulation has shown to be particularly reliable. The decisive factor, apart from any preferred method, is that the chosen therapeutic mandibular position be reproducible for both the treatment provider as well as the patient. PMID- 16708527 TI - Comments on medical qualification of space tourists. PMID- 16708526 TI - Medical qualification of a commercial spaceflight participant: not your average astronaut. AB - BACKGROUND: Candidates for commercial spaceflight may be older than the typical astronaut and more likely to have medical problems that place them at risk during flight. Since the effects of microgravity on many medical conditions are unknown, physicians have little guidance when evaluating and certifying commercial spaceflight participants. This dynamic new era in space exploration may provide important data for evaluating medical conditions, creating appropriate medical standards, and optimizing treatment alternatives for long-duration spaceflight. CASE: A 57-yr-old spaceflight participant for an ISS mission presented with medical conditions that included moderately severe bullous emphysema, previous spontaneous pneumothorax with talc pleurodesis, a lung parenchymal mass, and ventricular and atrial ectopy. The medical evaluation required for certification was extensive and included medical studies and monitoring conducted in analogue spaceflight environments including altitude chambers, high altitude mixed-gas simulation, zero-G aircraft, and high-G centrifuge. To prevent recurrence of pneumothorax, we performed video-assisted thoracoscopic pleurodesis, and to assess lung masses, several percutaneous or direct biopsies. The candidate's 10-d mission was without incident. CONCLUSION: Non-career astronauts applying for commercial suborbital and orbital spaceflight will, at least in the near future, challenge aerospace physicians with unknowns regarding safety during training and flight, and highlight important ethical and risk-assessment problems. The information obtained from this new group of space travelers will provide important data for the evaluation and in-flight treatment of medical problems that space programs have not yet addressed systematically, and may improve the medical preparedness of exploration-class missions. PMID- 16708529 TI - Acute cardiovascular adaptation to 10 consecutive episodes of head-up tilt. AB - BACKGROUND: The cardiovascular system is highly adaptable to sustained +Gz acceleration. Little is known as to whether the cardiovascular system can adapt to acute, repetitive +Gz exposures. This study tested the hypothesis that the cardiovascular system would adapt to repeated orthostatic challenges in a single session. METHODS: Over a 70-min period, 14 subjects were exposed to 10 +75 degrees head-up tilts (HUT). Each tilt involved a 5-min supine period followed by a 2-min HUT. Heart rate (HR), systolic BP (SBP), diastolic BP (DBP), mean arterial pressure (MAP), total peripheral resistance (TPR), stroke volume (SV), and cardiac output (CO) were determined non-invasively. Cardiovascular responses to HUT10 for the final 30 s of the supine period and the first 30 s of the tilt period were compared with those of HUT1. Integrated cardiac baroreflex sensitivity (BRS) was assessed using the Valsalva maneuver. RESULTS: MAP and DBP increased in both supine (MAP p = 0.009, DBP p = 0.002) and tilt periods (MAP p = 0.003, DBP p = 0.009) for HUT10 compared with HUT1. TPR increased during the tilt period only (p = 0.001) during HUT10 compared with HUT1. CO and SV were decreased during the supine period at HUT10 relative to HUT1; however, there were no differences in the tilt period at HUT10 for either CO or SV. There was no change in the response of BRS, HR, or SBP from HUT1 to HUT10. CONCLUSIONS: This study indicates that 10 repetitive HUTs can elicit changes in the cardiovascular responses to orthostasis, reflected by an increased vascular resistance. PMID- 16708528 TI - Inspiratory impedance effects on hemodynamic responses to orthostasis in normal subjects. AB - BACKGROUND: Breathing through an impedance threshold device (ITD) might prove effective as a countermeasure against post-spaceflight orthostatic hypotension since it increased blood pressure (BP) and cardiac output in supine human subjects. OBJECTIVE: We tested the hypothesis that spontaneous breathing through an ITD would attenuate the reduction in stroke volume and BP during orthostasis in human subjects. METHODS: There were 19 volunteers (10 men, 9 women) who completed two 80 degrees head-up tilt (HUT) protocols with (active) and without (sham control) an ITD set to open at -7 cm H2O pressure. Heart rate (HR), stroke volume (SV), cardiac output (CO), mean arterial pressure (MAP), and total peripheral resistance (TPR) were measured non-invasively during transition from supine to HUT. RESULTS: HUT caused significant elevation in HR and reductions in SV, CO, TPR, and MAP. Hemodynamic effects of HUT were similar for sham and active ITD. Further analysis revealed a subset (n = 11) of subjects who demonstrated a > 20% decrease in SV during HUT with the sham ITD. In this subset of subjects, the ITD attenuated (p = 0.004) the %deltaSV (-22.5 +/- 3.0%) during HUT compared with the sham ITD (%deltaSV = -37.4 +/- 2.6%). There was no statistical effect of ITD use in the subgroup who demonstrated < 20% reduction in SV (-16.6 +/- 0.4%). CONCLUSIONS: Use of an ITD may provide significant protection against orthostatic compromise in individuals with greater than 20% reductions in SV, such as astronauts returning from space. PMID- 16708530 TI - Semicircular canal influence on the visually perceived eye level during gondola centrifugation. AB - BACKGROUND: When exposed to an increased gravitoinertial force, a subject, sitting upright, experiences an illusion of being tilted backwards. This so called "G-excess illusion" is generally ascribed to the otolith organs. The present study aimed at elucidating how stimulation of the semicircular canals may influence the development of the G-excess illusion. METHODS: The visually perceived eye level (VPEL) was measured by means of a visual indicator in a large swing-out gondola centrifuge. The roll position of the gondola was controlled so that the subject was always upright with respect to the resultant vector of the Earth gravity force and the centrifugal force. Subjects (n = 8) underwent four centrifuge runs (2 G, 5 min), sitting in different positions, i.e., heading forwards, backwards, centripetally, and centrifugally. RESULTS: At the 2-G plateau there was a depression of the VPEL which was initially small but increased with a time constant of 90 +/- 30 s toward an asymptote of -22.0 +/- 6.9 degrees (mean and 1 SD for all positions). The initial depression was significantly smaller for the centripetal (+2.0 +/- 14.6 degrees) than for the centrifugal position (-14.5 +/- 10.4 degrees). However, there was no difference between the forward (-5.6 +/- 4.8 degrees) and backward (-4.0 +/- 4.5 degrees) positions. Initially after deceleration of the centrifuge to 1 G there was still a significant depression of the VPEL (-13.5 +/- 7.9 degrees), decreasing with a time constant of 100 +/- 46 s. CONCLUSIONS: The considerable delay in the otolith mediated changes in the VPEL is interpreted as due to the absence of adequate canal information for a change in head position. The difference in VPEL between the centripetal and centrifugal positions suggests an influence of canal change in-position information. However, pitch-plane angular velocity, being of considerable magnitude but of opposite sign for the forward and backward positions, did not influence the VPEL. PMID- 16708531 TI - Predicting neck injuries due to head-supported mass. AB - BACKGROUND: Technological advances in military equipment have resulted in more devices being mounted on the helmet to enhance the capability of the soldier. The soldier's neck must bear this head-supported mass (HSM) and the resulting dynamic characteristics of the head and neck system are changed. The purpose of this study was to vary the conditions of impact as well as the design criteria to quantify the effect of HSM on neck injury risk through computational modeling. METHODS: The TNO MADYMO detailed neck model was used for a matrix of 196 simulations designed to vary the impact conditions and HSM properties added to the model. These parameters included seven impact directions, three impact magnitudes, nine mass locations, and three mass magnitudes. The data collected from these simulations were evaluated for injury risk using the lower neck beam criterion equation. RESULTS: The results from these simulations provide detailed information regarding the risk of injury based on a particular HSM configuration and the acceleration of the body. The predominant factor in increasing risk in the lower neck is the increase in pulse magnitude. The effect of pulse magnitude is more dominant in the directions that create a flexion or lateral bending moment. CONCLUSION: HSM increases the level of injury, but the impact level that the subject is exposed to is a more dominating factor in determining injury risk. PMID- 16708532 TI - Prior sleep with zolpidem enhances the effect of caffeine or modafinil during 18 hours continuous work. AB - INTRODUCTION: Continuous military operations may disrupt sleep-wakefulness cycles, resulting in impaired performance and fatigue. We assessed the treatment efficacy of a hypnotic-psychostimulant combination to maintain sleep quality, performance, and alertness during a 42-h simulated military operation. METHODS: A 6-h prophylactic sleep period with zolpidem (ZOL) followed by a 18-h continuous work period with administration at midway of 300 mg of slow release caffeine (CAF) or 200 mg of modafinil (MOD) was performed by eight healthy male subjects. Performance level was assessed with a reaction time test, a memory search test, a dual task, an attention test, and a computerized Stroop test. Cortical activation level was evaluated by the Critical Flicker Frequency test. Subjective sleepiness was evaluated using a visual analog scale and questionnaires. Effects of drugs on prophylactic and recovery sleep were also quantified from EEG recordings. RESULTS: CAF and MOD maintained performance and alertness throughout the 18-h work period. As shown by EEG recordings, ZOL improved prophylactic sleep without any deleterious effect on performance immediately after waking. As a result of its positive effects on prophylactic sleep, a lower pressure for slow wave sleep during recovery sleep was observed; nevertheless, zolpidem did not enhance the effects of either psychostimulant on performance. DISCUSSION: MOD and CAF may be of value in promoting performance and wakefulness during shiftwork or military operations while zolpidem improves prophylactic sleep quality without any deleterious effect after waking. We concluded that a zolpidem/ caffeine or modafinil combination could be useful in a context of environmental conditions not conducive to sleep. PMID- 16708533 TI - CNS toxicity in closed-circuit oxygen diving: symptoms reported from 2527 dives. AB - INTRODUCTION: Oxygen toxicity is a problem in diving and can have fatal consequences in the water. Various aspects of oxygen diving have been studied in dry hyperbaric chambers, but there is a lack of information on in-water diving using closed-circuit oxygen apparatus. METHOD: We collected 2527 dive reports from 473 closed-circuit oxygen divers (a mean of 5.2 reports per diver), and analyzed the relationships between various symptoms and their dependence on depth and diving time. RESULTS: No CNS oxygen toxicity-related symptoms were reported at a depth of 2 m seawater (msw), but their proportion increased at depths from 3 to 6 msw. We found that CNS oxygen toxicity-related symptoms appeared in 2.5% of dives conducted at a Po2 of 119 kPa. The main symptoms and signs reported were headache: 4.5%; nausea: 2.6%; hyperventilation: 2.6%; heavy breathing: 2.4%; dizziness: 1.6%; hiccups: 1.5%; bloody sputum: 1.4%; cold shivering: 1.1%; tinnitus: 0.9%; difficulty maintaining a steady depth: 0.9%; disorientation: 0.6%; tiredness: 0.5%; tingling in the limbs: 0.4%; hearing disturbances: 0.4%; a choking sensation: 0.4%; extreme effort: 0.4%; and loss of consciousness: 0.3%. DISCUSSION: Environmental factors, light vs. dark and temperature, had no effect on symptoms. The number of symptoms increased with diving time. Divers who experienced amnesia, facial twitching, hearing disturbances (p < 0.001), and disorientation (p < 0.014) were prone to suffer loss of consciousness. It was found that some divers are more sensitive to oxygen than others (p < 0.0001). PMID- 16708534 TI - Reproductive disorders and pregnancy outcomes among female flight attendants. AB - BACKGROUND: Occupational exposures in female aircrew may cause adverse pregnancy outcomes and menstrual disturbances. We studied reproductive health among female flight attendants. METHODS: We conducted a cross-sectional health survey among an occupational cohort of current and former flight attendants using a postal questionnaire including items on pregnancy outcome, menstrual characteristics, and infertility. We investigated these factors by occupational status (in service or not). RESULTS: The questionnaire was sent to 3036 women with a response rate of 64% (74% for current and 48% for former flight attendants). Spontaneous abortion rates were similar for pregnancies of women in service and not (12.6 vs. 11.4%; p = 0.58). Induced abortion rates were lower for in-service pregnancies (7.9%) compared with pregnancies of women not in service (21.1%) (p < 0.001). Menstrual irregularities in the year preceding the survey for women under 40 yr were more frequent among current than former flight attendants (20.6% vs. 10.4%, p = 0.02). Fertility problems were reported by 20.6% of respondents. An association between infertility and irregular menstrual cycles was found: odds ratio 1.6; 95% confidence limits 1.1, 2.4. CONCLUSIONS: Active flight attendants had a lower rate of induced abortions than former flight attendants, but the spontaneous abortion rate was similar between the two groups. Active flight attendants reported more menstrual irregularities, which are a risk factor for infertility. Lack of comparison with working women in other occupations precludes a conclusion that flight attendants are at greater occupational risk of reproductive disorders. Future studies of reproductive health in flight attendants should address personal and work-related risk factors in more detail and include comparison occupational groups. PMID- 16708535 TI - Heat stress reduction of helicopter crew wearing a ventilated vest. AB - BACKGROUND: Helicopter pilots are often exposed to periods of high heat strain, especially when wearing survival suits. Therefore, a prototype of a ventilated vest was evaluated on its capability to reduce the heat strain of helicopter pilots during a 2-h simulated flight. HYPOTHESIS: It was hypothesized that the ventilated vest would reduce pilot heat stress. METHODS: Five male and one female helicopter pilots flew for 2 h in a simulator in three different conditions; 15 degrees C wet bulb globe temperature (WBGT) without ventilation, 32 degrees C WBGT without ventilation, and 32 degrees C WBGT with a ventilated vest. RESULTS: Wearing the ventilated vest significantly reduced the increase in rectal temperature and increased thermal comfort. This made it possible for all subjects to complete the 2-h session. CONCLUSION: With the ventilated vest the subjects experienced less heat stress, thereby allowing all subjects to successfully complete the experiment, though two of the six pilots could not complete the 2-h flying task in the hot condition without cooling due to heat-related problems. PMID- 16708536 TI - Plasma glutathione peroxidase activity as a potential indicator of hypoxic stress in breath-hold diving. AB - INTRODUCTION: Diving mammals can cope with oxidants which are produced in excess during the reoxygenation of hypoxic tissues. This study addresses the question of whether antioxidants can adapt and whether it allows humans to tolerate the hypoxic stress induced by a single breath-holding in the course of a dynamic diving exercise and protect them from oxidative insult. METHODS: There were 20 male subjects who performed submaximal apnea dynamic diving (ADD). Nine control subjects stayed out of the water and breathed normally. Venous blood samples were collected 1 h before and immediatly after ADD. RESULTS: ADD induced a significant increase in plasma glutathione peroxidase (GPx-3) activity (from 397.5 +/- 44.4 to 410 +/- 43 U x L(-1)), blood reduced glutathione (GSH) (from 1060 +/- 302 to 1292 +/- 213 micromol x L(-1)), and in plasma creatine kinase activity (from 215 +/- 137 to 235 +/- 152 U x L(-1)). The activity of the erythrocyte superoxide dismutase and glutathione peroxidase, as well as the blood oxidized glutathione and the plasma thiobarbituric acid reactive substances concentrations, were maintained at their basal level. The level of training, characterized by the duration and distance of the dive, had no effect on the markers used. CONCLUSION: GPx-3 and GSH could constitute the most readily mobilizable antioxidants that would then contribute to the buffering against a sudden increase in the generation of radical oxygen species. These biomarkers could be used as tools for establishing oxidative stress during hypoxia. The response of GPx-3 to hypoxia could be of physiological relevance. PMID- 16708537 TI - Balloon crash damage and injuries: an analysis of 86 accidents, 2000-2004. AB - BACKGROUND: General aviation accounts for the majority of aviation crashes and casualties in the United States. The role of ballooning in these statistics is not regularly studied. Since 2001, the National Transportation and Safety Board has made its accident reports more readily available, which presents opportunities for further study. OBJECTIVE: This study analyzes and compares a 5 yr period of accident reports and includes an analysis of injuries and balloon damage in hot-air and gas balloon accidents. METHODS: Balloon crash 2-page briefs and 5-page accident reports published by the National Transportation and Safety Board for the 5-yr time period 2000-2004 were examined. Data collected in the investigation of these crashes were analyzed and compared with the epidemiological data collected in earlier research. RESULTS: In 86 crashes during a 5-yr period, there were 4 fatalities and 75 people were seriously injured. Only one accident was reported involving a student pilot. Broken ankles and legs have been the most commonly recorded serious injury, but could not be linked to the severity of damage to the balloon. CONCLUSIONS: The absence of student pilot accidents may be explained by possible stricter supervision. Balloon basket and envelopes appear of sufficient quality to withstand crashes, but improving the protection of passengers during hard landings should help to decrease the number of serious injuries in ballooning. PMID- 16708538 TI - You're the flight surgeon: a retinal hole with operculum. PMID- 16708539 TI - At your service--results of the readership survey. PMID- 16708541 TI - This month in aerospace medicine history--May 2006. PMID- 16708540 TI - Digital signal processing actigraphy. PMID- 16708542 TI - Expression patterns of 5-lipoxygenase in human brain with traumatic injury and astrocytoma. AB - 5-Lipoxygenase (5-LOX) is a key enzyme in the metabolism of arachidonic acid to leukotrienes. The levels of leukotrienes increase after brain injury and when tumors are present. It has been reported that 5-LOX is widely expressed in the brain and that 5-LOX inhibition provides neuroprotection. However, there is still no information available for the expression patterns of 5-LOX in human brain following trauma or with astrocytomas. We investigated its expression patterns by immunohistochemistry. We found that 5-LOX is normally expressed in neurons and glial cells. In neurons, it was expressed in two patterns: in the cytosol and nucleus or only in the cytosol. In traumatic brain injury, 5-LOX expression increased in glial cells and neutrophils. Double-labeling immunohistochemistry showed that part of the 5-LOX-positive glial cells were GFAP positive. No 5-LOX expression was found in brain microvessel endothelia, except in the regenerated endothelia of a patient 8 days following brain trauma. Furthermore, 5-LOX expression increased and showed a granular pattern in high-grade (grade III/IV) astrocytoma. These results indicate that 5-LOX has multiple expression patterns, and can be induced by brain injury, which implies that 5-LOX might have pathophysiological roles in the human brain. PMID- 16708543 TI - Neurofibrillary tangles and deposition of oxidative products in the brain in cases of myotonic dystrophy. AB - Myotonic dystrophy (MyD) is a neuromuscular degenerative disorder that is neuropathologically characterized by minor changes, such as the presence of neurofibrillary tangles (NFT), thalamic inclusions and functional brainstem lesions. In the current study, we conducted an immunohistochemical analysis to examine the distribution of NFT and formation of oxidative products in the brain specimens of 12 patients with MyD. Neurofibrillary tangles were found in the limbic system and/or the brainstem of all the cases examined but there were no senile plaques. The density of distribution of the NFT was not significantly correlated with clinicopathological findings, although cases with fewer NFTin the brain frequently showed sleep disturbances and lack of spontaneity. Nuclear and cytoplasmic immunoreactivities for 8-hydroxy-2'-deoxyguanosine and advanced glycation end products were observed in the glial cells and/or neurons in the brainstem, but not in the cerebral cortex. On the other hand, 10 out of the 12 cases showed cytoplasmic immunoreactivity for 4-hydroxy-2-nonenal-modified protein (4-HNE) in neurons of the temporal cortex and raphe nucleus. Deposition of 4-HNE was also recognized in the hippocampus and mesencephalic central gray matter, but not in the subiculum. The distribution pattern of the immunoreactivity for 4-HNE showed no clear correlation with either the psychological disturbances or the distribution of the NFT. Altered expression of monoaminergic neurons in the brainstem of MyD patients has already been reported, and it is worth noting that most of our cases showed NFT in the brainstem. The selective deposition of 4-HNE in the limbic system and brainstem suggests that lipid peroxidation may be involved in the neurodegenerative process in MyD. Using immunohistochemical analysis to determine the distribution of neurotransmitters in the mesencephalic central gray matter and/or pontine raphe nucleus may help elucidate the relationship between the clinical abnormalities, distribuion of NFT, and 4-HNE deposition in the brain in patients with MyD. PMID- 16708544 TI - Magnesium deficiency over generations in rats with special references to the pathogenesis of the Parkinsonism-dementia complex and amyotrophic lateral sclerosis of Guam. AB - Parkinsonism-dementia complex (PDC) and amyotrophic lateral sclerosis (ALS) are fatal neurological diseases. The incidence on Guam was very high between 1950 and 1965 but decreased dramatically after 1965. It is thought that drinking water containing low levels of calcium (Ca) and magnesium (Mg), and high levels of aluminum and of a plant excitatory neurotoxin are involved in the pathogenesis of these diseases. The present experiment was performed in rats that were exposed to low Ca and/or Mg intake over two generations, thus simulating the conditions of human life on Guam, where several generations live continuously in the same environment. Significant loss of dopaminergic neurons was identified exclusively in the substantia nigra in 1-year-old rats that had been exposed continuously to low Mg intake (one-fifth of the normal level) over generations. The present study suggests that low Mg intake over generations may be involved in the pathogenesis of substantia nigra degeneration in humans. PMID- 16708546 TI - Benign osteoblastoma of the occipital bone: case report and literature review. AB - We present a case of benign osteoblastoma of the occipital bone. Benign osteoblastoma is an uncommon primary bone tumor, which usually involves the vertebrae and the long bones. This tumor rarely develops in the calvaria, showing a preference for the temporal and frontal bones when it does. To the best of our knowledge, this case is only the eighth reported case of benign osteoblastoma confined to the occipital bone. A 20-year-old male presented with a mild tender mass lesion of the occipital area, just below the lambda. Plain X-ray films and CT scans demonstrated an osteolytic mass surrounded by the sclerotic rim within the diploic space. MRI proved to be effective for the evaluation of the intracranial and intraosseous extensions of the tumor. However, it was very difficult to formulate a differential diagnosis against other osteoblastic tumors, or osteoid osteoma, in view of its radiological appearance. The final diagnosis was obtained by careful consideration of the histopathological characteristics of the tumor combined with its clinical and radiological features. Although generally regarded as benign, a complete resection is preferred over conventional curettage as this can guard against possible recurrence and malignant transformation. PMID- 16708545 TI - Brain transplantation of human neural stem cells transduced with tyrosine hydroxylase and GTP cyclohydrolase 1 provides functional improvement in animal models of Parkinson disease. AB - Parkinson disease is a neurodegenerative disease characterized by loss of midbrain dopaminergic neurons resulting in movement disorder. Neural stem cells (NSC) of the CNS have recently aroused a great deal of interest, not only because of their importance in basic research of neural development, but also for their therapeutic potential in neurological disorders. We have recently generated an immortalized human NSC cell line, HB1.F3, via retrovirus-mediated v-myc transfer. This line is capable of self-renewal, is multipotent, and expresses cell specific markers for NSC, ATP-binding cassettes transporter (ABCG2) and nestin. Next, we co-transduced the F3 NSC line with genes encoding tyrosine hydroxylase (TH) and GTP cyclohydrolase 1 (GTPCH1) in order to generate dopamine-producing NSC. The F3.TH.GTPCH human NSC line expresses TH and GTPCH phenotypes as determined by RT PCR, western blotting and immunocytochemistry, and shows a 800 to 2000-fold increase in production of L-dihydroxyphenyl alanine in HPLC analysis. A marked improvement in amphetamine-induced turning behavior was observed in parkinsonian rats implanted with F3.TH.GTPCH cells, but not in control rats receiving F3 NSC. In the animals showing functional improvement, a large number of TH-positive F3.TH.GTPCH NSC were found at injection sites. These results indicate that human NSC, genetically transduced with TH and GTPCH1 genes, have great potential in clinical utility for cell replacement therapy in patients suffering from Parkinson disease. PMID- 16708547 TI - Hypoglycemic encephalopathy with extensive lesions in the cerebral white matter. AB - Here we report an autopsy case of hypoglycemic encephalopathy with prolonged coma. Laboratory data obtained when the patient lapsed into a coma showed that she had a low level of serum glucose (27 mg/dL). Although the level of glucose returned to within the normal range rapidly after glucose infusion, the patient remained in a coma for 22 months. It was presumed that the state of hypoglycemia persisted for about 4 h. There was no evidence of hypotension or hypoxia. Magnetic resonance imaging was performed 3 h after glucose administration; diffusion-weighted images revealed hyperintensity in the cerebral white matter and in the boundary zone between the middle and posterior cerebral arteries. Post mortem examination revealed superficial laminar necrosis throughout the cerebral cortex. Neuronal necrosis was also found in the hippocampus and dentate gyrus, although the CA3 region appeared normal. In addition to these lesions, which are consistent with hypoglycemia-induced brain damage, the cerebral white matter exhibited severe loss of myelin and axons with reactive astrocytosis and macrophage infiltration. Old infarcts were also present in the bilateral occipital lobes. Since the cerebral blood flow is reported to be decreased during severe hypoglycemia, the present findings suggest that white matter lesions and boundary-zone infarctions may develop primarily in uncomplicated hypoglycemia. PMID- 16708548 TI - Pineal parenchymal tumors of intermediate differentiation in adults: case report and literature review. AB - Pineal parenchymal tumors (PPT), including pineoblastomas, are very uncommon, especially in adults. Because of the small number of reported cases, the histological and biological features of these tumors are still being defined, as is their optimal management. Also, the pathological variability of these tumors makes it difficult to draw general conclusions about their behavior. We report the case of a PPT of intermediate differentiation (PPTID) with an excellent outcome. A 3.2 cm tumor in the pineal and tectal region with obstructive hydrocephalus was found in a 28-year-old female complaining of headaches and hypoesthesia of the right side of the face. Stereotactic biopsy revealed a highly cellular tumor composed of small cells that contained little cytoplasm that were arranged in a diffuse pattern. Mitotic activity was low and there was no evidence of necrosis. Immunohistochemical examination demonstrated positive staining for neuron-specific enolase and synaptophysin. There was no expression of neurofilaments or GFAP. Ki-67 proliferation index was 12%. The diagnosis was a PPTID. Treatment consisted of combined neoadjuvant chemotherapy and craniospinal radiotherapy. Six years after treatment, the patient is alive and free of any clinical or radiological signs of relapse. This paper discusses the pathological and biological features of these tumors and the treatment options available. PMID- 16708549 TI - Fine structural study of a cerebral tumor characterized by a honeycomb appearance after a 20-year post-mortem interval. PMID- 16708550 TI - How AIDS changed America. PMID- 16708551 TI - Battling a black epidemic. PMID- 16708552 TI - A challenge for leaders. PMID- 16708553 TI - My quest to improve care. PMID- 16708554 TI - The life of a virus hunter. PMID- 16708555 TI - An HIV survivor's story. PMID- 16708556 TI - What women really need. PMID- 16708557 TI - [The influence of IL-6 polymorphism on efficacy of treatment of rheumatoid arthritis patients with methotrexate and prednisone]. AB - Rheumatoid arthritis (RA) is a chronic inflammatory disease in which cytokines play an important role. The therapy of RA is associated with application of the drugs modulating the immune response via inhibiting the cytokine production. The common drugs used in RA therapy are methotrexate and prednisone. Recent investigations showed the importance of genetically determined differences in cytokine production in RA activity and therapy. The aim of the present study was to examine the influence of - 174 interleukin-6 (IL-6) promoter polymorphism on the efficacy of treatment of RA patients with methotrexate and prednisone. Polymerase chain reaction amplification was used for analysis of the polymorphism at IL-6 gene. Seventy patients with RA diagnosed according to the criteria of the American College of Rheumatology were investigated. The patients were divided into two subgroups. The first subgroup included patients who have obtained remission for at least 6 months after therapy with methotrexate and glucocorticosteroids. The second subgroup included patients with active disease despite at least 6 months of therapy with methotrexate and glucocorticosteroids. It has been shown that the incidence of remission after therapy with methotrexate and glucocorticosteroids was significantly lower in patients with GG genotype as compared with GC and CC genotypes p< 0.05. We suggest that -174 IL-6 promoter polymorphism may be a genetic risk factor determining the effectiveness of RA treatement with methotrexate and glucocorticosteroids. PMID- 16708558 TI - [Patterns of body surface isointegral maps for anteroseptal and inferior myocardial infarction]. AB - A goal of the present paper was to determine the patterns of the QRS isointegral maps for the two location of myocardial infarction: anteroseptal (aMI) and inferior (iMI), using a method of body surface potential mapping (BSPM) with a 87 electrode Fukuda Denshi system. The maps were recorded in the two groups of the patients with previous (6-12 month earlier), clinically documented, Q-wave myocardial infarction. The examined group comprised 36 patients with aMI and 32 patients with iMI. The analysis concerned the isointegral maps of the assigned seven time intervals within the QRS complex (the A-G maps), rendering the patterns of positive and negative potential distribution, likewise the group-mean values of minima and maxima for each of the analyzed maps. The increased area of negative potential, as compared with the corresponding control maps, was observed in the maps of the A, B and D intervals in the aMI group. Contrary, in the iMI group a pathological negative potential was found only in the E maps (the second half of the QRS complex). The comparative analysis of the potential extremes revealed in the aMI group the significantly more negative minima in the A, B, D and E maps and lower maxima in the A and B maps. However, in the iMI group the only statistically significant difference were the lower minima for the E maps. The investigations resulted in creating the patterns of the pathological distribution of the negative potential and the minimum values in the isointegral QRS maps, which are specific for the anteroseptal and inferior myocardial infarction. PMID- 16708559 TI - [Heart rate variability in patients with chronic renal failure treated by hemodialysis]. AB - Heart rate variability (HRV) is a non-invasive method used for the assessement of autonomic modulation of heart rate. Decreased HRV is an indicator of increased cardiovascular risk. The aim of this study was to evaluate the relationship between the heart rate variability and left ventricular hypertrophy and native parathormone (iPTH) serum concentration in patients with chronic renal failure (crf) treated by hemodialysis. 24-hours ECG recording with time domain HRV evaluation, resting, transthoracic echocardiography (ECHO), were measured in 59 crf patients and in 30 healthy volunteers. Creatinine, urea, total protein, albumin, electrolytes, hemoglobin, hematocrite and iPTH serum concentration as well as body mass index (BMI) were assessed in all patients. All crf patients had decreased lower values of HRV. The correlations between SDNN, pNN50, rMSSD and parameters of LVH and with PTH serum level indicated the disturbances of the autonomic function in chronic renal patients. Left ventricular hypertrophy (LVH) in all crf patients was observed. The correlations between iPTH serum level and parameters of LVH suggest the role of PTH in the development of uremic cardiomyopathy. PMID- 16708560 TI - Acetylsalicylic acid (ASA) use among Polish adults with diabetes mellitus. AB - People with diabetes mellitus have higher risk of cardiovascular morbidity and mortality from thrombo-vascular complications than non-diabetics and it is recommended that they should use acetylsalicylic acid (ASA) as anitiplatelet agent regularly. The aim of this study was to examine current physician counseling about ASA therapy and the use of ASA by Polish patients with diabetes. The study involved 464 randomly selected Caucasian diabetic individuals, mainly with type 2 (>98%), who were asked to complete a questionnaire consisting of 16 questions while visiting out-patient medical centres for periodical examination or during hospitalization. We collected data on the prevalence of cardiovascular disease (CVD), frequency of risk factors among diabetic patients and on ASA physician counseling, and on the rate of regular ASA users among the patients. The most self-reported conditions were angina pectoris or prior myocardial infarction (63,6%). Current cigarette smoking was declared by 7,1% of the patients, elevated serum cholesterol levels by 58%, and hypertension by 62,9%. Majority of the patients were overweight, and family history of coronary artery disease (CAD) was reported by 30,2% of the patients. ASA was taken by 67,2% of all the patients and 72,1% of them had taken ASA regularly. The highest percentage (>80%) of regular ASA users was found among diabetic patients with CAD. A minority of the patients who reported stroke (43,9%) or claudication (15,8%) were taking ASA regularly. Number of the patients (32,8%) with CVD risk factor(s) but without diagnosed CVD did not use ASA at all. The majority of the patients (78%) were counseled by their physicians why they should use ASA regularly. IN CONCLUSION: the number of regular users of ASA among diabetic patients in Poland with CVD others than CAD or among patients with at least two CAD risk factors is still unsatisfactory. Therefore, while it seems that there is a remarkable growing of ASA therapy world-wide more effort is needed to make this practice a standard of multifactorial, intensive diabetes treatment. Our results also suggest the need for more aggressive education of both doctors and patients. PMID- 16708562 TI - [Quality of life in patients after the Ross operation for aortic stenosis]. AB - The pulmonary autograft provides ideal conditions regarding hemodynamics. Its additional value is durability and low thromboembolic hazard. We postulated that the Ross technique results in minimal restrictions on patient's lifestyle as compared to normal controls and we expected the patients to have good quality of life (QoL). On the other hand QoL may be affected by the risk of the homograft degeneration and the need of reoperation in the future. The aim of the study was to assess the QoL of patients after Ross operation because of aortic valve stenosis. Postoperative condition of the patients was judged by resting transthoracic echo, 24 hour ECG recording, maximal exercise test on bicycle ergometer. The mean age of patients was 31.1 (N=50; 18 female, 32 male). The mean follow-up was 3.6 years. We applied the Short Form Health Survey Questionnaire (SF-36) for assessment of QoL. The results of psychological tests were compared with age, gender and education matched control group without underlying cardiac disease. Overall QoL (QoL index) did not vary between patients and control group. The only significant difference concerned physical sum score and subtests of physical functioning (scale 1) and general health perception (scale 8). There were no differences in psychological health sum score. We concluded that in general, patients after the Ross operation enjoy a normal life style. The results of our study are consistent with well known advantages of Ross procedure. This study also implies the need for more detailed information concerning patient's lifestyle after the operation. Patient's family and doctors who are to take care of the patient after leaving hospital ought to be instructed as well. Education should specify patient's ability to perform different everyday abilities, especially physical ones. The aim of the education is to eliminate negative self assessment of patient's own physical abilities which probably originate from preoperative period and stress connected with operation. PMID- 16708561 TI - [Telmisartan in monotherapy of essential hypertension in young men--time of drug administration and 24-hours blood pressure and heart rate]. AB - 42 men, aged 19-35 years (mean 26.3), with mild or moderate essential hypertension (DBP 95-110 mmHg), were treated with telmisartan (40 mg or 80 mg ) once daily in the morning (6 and 7 a.m.) for at least 6 weeks and for next 6 weeks and after that administration time was changed to evening dose (6-7 p.m.) 24-hours ABPM (SpaceLabs) was performed in all patients on last day of each period. Obtained recordings were compared in different periods of time: 6.00 a.m. -10.59 p.m., 11.00 p.m-5.59 a.m., 2.00-5.59 a.m., 6.00-12.00 a.m, 3.00-7.00 p.m. The SBP values in all time intervals were comparable (differences statistically not significant). A statistically significant reduction of the DBP in the time interval 6:00-12:00 a.m. was found in patients treated with telmisartan in the morning. The heart rate of the patients who received telmisartan has not been affected by the treatment schedule and remained comparable. CONCLUSIONS: 1.) The time of the day administration does not affect the diurnal and night hypotensive efficacy of telmisartan. 2.) According to the results of this study, patients with the essential hypertension should receive telmisartan as monotherapy in the morning. PMID- 16708563 TI - [Nephrogenic diabetes insipidus induced by colchicine--a case report]. AB - In some cases diabetes insipidus may be induced by taking medications. In this article we report the case of a 58-year-old man with manifest polyuria induced by colchicine administered because of a gout attack. Interestingly, similar symptoms were observed 6 years earlier after treatment with gentamycin. In the described state of the examined patient early detection of the disease, colchicine withdrawal and temporary amiloride (and indometacin) administration led to a full normalisation of the patient's clinical status. A similar reaction to two different drugs responsible for only a small percentage of iatrogenic diabetes insipidus suggests that some patients may exhibit a general "hypersensitivity" to the development of a drug-induced vasopressin resistance and that patients with the history of iatrogenic diabetes insipidus should not be treated or should be treated cautiously with other drugs known to induce vasopressin-resistant polyuria. PMID- 16708564 TI - [Kidney sponge--treatment--case report]. AB - Medullary sponge kidney (MSK) is a benign asymptomatic developmental anomaly of the kidney mostly seen in adult females. Typical for this morphological abnormality is dilation of the collecting ducts. Intravenosus pyelogram shows accumulation of contrast in dilated ducts giving to the papillae the appearance of a bouquet flowers, characteristic for MSK. Urinary tract infections, nephrolithiasis, hematuria and hyperkalciuria are the common complications of the kidney sponge. We present a case of a 29-year-old female who suffers from recurrent urinary tract infection, nephrolithiasis and distal tubular acidosis. This kind of tubular acidosis is specific for kidney sponge clinical picture. PMID- 16708565 TI - [Single nucleotide polymorphism of protein tyrosine phosphatase 1B (PTP-1B) gene. Clinical implications in type 2 diabetes]. PMID- 16708566 TI - [Adrenal cortical carcinoma. Progress in diagnosis, clinical and genetic features]. PMID- 16708567 TI - [Is it recommended simultaneous use of acetylsalicylic acid and angiotensin converting enzyme inhibitors?]. PMID- 16708568 TI - [Fever of unknown origin]. PMID- 16708569 TI - [Mechanisms and medical implications of replicative senescence]. AB - Normal somatic cells divide in vitro only for a limited number of times, and then enter a state of replicative senescence. In most human cells replicative senescence is triggered by critical shortening and uncapping of telomeres, which leads to the up-regulation of p53 and/or p16 suppressor proteins that inhibit cell divisions. Because both reconstruction of telomeres and inactivation of suppressor proteins enables the cells to grow further or even immortalize, it has been hypothesized that replicative senescence acts as a natural barrier against neoplastic transformation. On the other hand, however, recent reports suggest that the accumulation of senescent cells may jeopardize tissue integrity and create a specific microenvironment that promotes tumorogenesis in elderly people. In this review we describe recent advances in our knowledge on the mechanisms of replicative senescence and discuss its medical implications. PMID- 16708570 TI - [Calcific valvular heart disease in patients with aortic and peripheral atherosclerosis]. PMID- 16708571 TI - [The 111th congress of the German Society of Internal Medicine, Wiesbaden, April 2-6, 2005]. PMID- 16708572 TI - Discharge planning and the dilemma of dealing with homeless patients. PMID- 16708573 TI - Network targets barriers for unfunded patients. PMID- 16708574 TI - Approach increases reimbursement, cuts LOS. PMID- 16708576 TI - Case managers help mentally ill avoid hospitalization, remain in the community. PMID- 16708575 TI - Hospital funds, donations provide needed care. PMID- 16708577 TI - Program keeps unfunded patients out of the hospital. PMID- 16708578 TI - Hospital uses bright blue to nurture bottom line. PMID- 16708579 TI - Get a handle on your discharge process. PMID- 16708580 TI - Will centers make money with the move to APCs? PMID- 16708581 TI - [Extracorporeal shock-wave lithotripsy in the treatment of urolithiasis of dystopic kidneys]. AB - The 10-year experience of the urological clinic of M.F. Vladimirsky Research Clinical Institute included treatment of 219 patients (age 21-68 years, mean age 52.3 +/- 12.4; 59.2% females, 40.8% males) with urolithiasis which developed in the presence of anomalies of the kidneys and upper urinary tract. Uroliths were detected in the kidneys with homolateral (n = 51, 23.7%), lumbar (n = 49), iliac (n = 1), pelvic (n = 2) dystopia. Chest and heterolateral dystopia of the kidneys were not detected. Extracorporeal shock-wave lithotripsy (ESWL) was used in 38 (77.6%) patients, 11 (22.4%) patients were treated by other methods. A short-term effect of ESWL consisted in elimination of most of the concrement fragments while the residual fragments had no clinical symptoms being maximally 4-5 mm in size. It was noted in 23 of 38 (60.5%) patients. Six months after ESWL elimination of the concrement from lumbar-dystopic kidney occurred in 27 (71.05%) of 38 patients. Residual fragments 6-10 mm in size were registered in 26.35% patients, more than 10 mm--in 1 (2.6%). Complications after ESWL monotherapy of the concrements of the lumbar-dystopic kidney were seen in 26.3% (acute pyelonephritis 10.6%, stone track--7.9%, total hematuria--2.6%, acute prostatitis -2.6%, renal colic--2.6%). Mean treatment duration was 15.4 +/- 3.3 days. Thus, most effective therapy ofdystopic kidneys urolithiasis is ESWL the total efficacy of which reached 71.05%. PMID- 16708582 TI - [Urolithiasis: current aspects of diagnosis and treatment]. AB - The aim of the study was clinicodiagnostic assessment of current methods of computer visualization (spiral computed tomography, multispiral computed tomography) and their role in specification of indications, contraindications and technical characteristics of current methods of urolithiasis treatment. The results of combined examination and therapy of 759 urolithiasis patients treated in the urological clinic of I.M. Sechenov Moscow Medical Academy in 1999-2003 are presented. PMID- 16708583 TI - [Combined extract of Sabal palm and nettle in the treatment of patients with lower urinary tract symptoms in double blind, placebo-controlled trial]. AB - A multicenter, prospective clinical trial was performed to study efficacy and tolerance of a compound drug PRO 160/120 in the elderly men with lower urinary tract symptoms (LUTS) due to benign prostatic hyperplasia (BPH). A total of 257 patients were randomized into two groups. Group 1 of 129 patients received PRO 160/120; group 2 of 128 patients received placebo. In 2-week induction blind phase of placebo the patients received for 24 weeks 1 capsule of the drug or placebo twice a day in conditions of double blind study. The double blind phase was followed by an open control period for 24 weeks when all the patients received PRO 160/120. Treatment efficacy evaluation was based on I-PSS, quality of life index, urodynamic and ultrasonography evidence. PRO 160/120 was superior to placebo by attenuating LUTS assessed by I-PSS, improved obstructive and irritative symptoms, was effective in patients with moderate and severe symptoms. Tolerance of the plant extract was good. PMID- 16708584 TI - [Fluorescence detection of photosensitizer photoditazin in the tissue of benign prostatic hyperplasia]. AB - The authors present pilot experience in investigation of accumulation, distribution and elimination of photosensitizer photoditazin from hyperplastic tissue of human prostate. Fluorescent spectroscopy showed that 2-24 hours after intravenous injection of photoditazin in a dose 1 mg/kg, it is detected in hyperplastic tissue of human prostate with maximal accumulation 3 hours after introduction. Exogenic fluorescence of photoditazin lowered 12 and 24 hours after injection this indicating its elimination. The results favor use of photoditazin for photodynamic treatment of patients with benign prostatic hyperplasia. PMID- 16708585 TI - [Postoperative urological complications of transurethral electrosurgical interventions on the prostate for adenoma]. AB - The aim of the study was comparison of urological complications after transurethral resection and its low-invasive alternatives: vaporization, rotoresection, vaporization resection, prostatic incision. Case histories were studied of 5401 patients operated endoscopically for prostatic adenoma in 1991 2003. Standard TUR was made in 5003 patients, incision--in 112, vaporization--in 119, vaporizing resection--in 107, rotoresection--in 60 patients. Of early complications after TUR hemorrhage was the most dangerous, hemotransfusion was made in 3.9% cases. No hemorrhagic complications occurred in rotoresection, no hemotransfusions were made in vaporization and vaporizing resection, prostatic incision was complicated by hemorrhage only in 0.9% cases. By infection complications rate (0.9-7.5%), the differences were not significant. Of late complications, sclerosis of the urinary bladder cervis was most frequent (10.7 11.2%) after rotoresection and vaporizing resection, urethral stricture--after TUR (6.9%). By number of complications, incision was least invasive. Thus, basic problem after TUR is hemorrhage. Low-invasive alternative methods resolve this problem. But they have limitations. The best way out is improvement of intraoperative hemostasis during TUR. PMID- 16708586 TI - [Conservative treatment of Peyronie's disease in the light of new pathogenetic data]. AB - The study of pathogenetic mechanisms of Peyronie's disease (PD) in respect of raising efficacy of complex conservative therapy was made in 41 patients with PD. The control group consisted of 266 persons. Immunological and virusological investigations were made using enzyme immunoassay. HLA typing of class I antigens was made, antinuclear antibodies and antibodies to DNA were investigated. Effects of intron A and verapamil on proliferation of penile tunica albuginea were studied in vitro. Conservative therapy was given to 32 patients with acute PD. Patients with PD were found to carry chronic infection with type II herpes significantly more frequently. There was a significant association of PD with HLA antigen B8 with high percentage of diagnostic titers of antinuclear antibodies. In vitro effect of intron A and verapamil was found to be dose- and time dependent. Verapamil has a narrow range of dose-dependence and cytotoxicity in high concentrations. Combined treatment raised the proportion of good results while that of satisfactory outcomes decreased. Viral infection may be involved in pathogenesis of PD. This infection may alter mechanisms of immune regulation and start of autoimmune process in predisposed patients. Combination of magnetolaser therapy with intron A injections is an effective method of acute PD treatment. The addition of specific antiviral therapy raises treatment efficacy by action on one of pathogenetic mechanisms of the disease. PMID- 16708587 TI - [Comparative assessment of combined therapy of neurogenic hyperactivity of detrusor in patients with Parkinson's disease]. AB - We studied clinical characteristics of voiding disorders to improve treatment results in patients with Parkinson's disease (PD) in 64 PD patients (28 women and 36 men aged 45 to 76 years, mean age 62.5 +/- 9.4 years). Duration of PD was 8.0 +/- 3.4 years, history of voiding disorders--2.1 +/- 0.8 years. Hyperactive urinary bladder was the cause of voiding disorders in all the patients. The examinees have undergone physical examination, microscopic and cultural examination of the urine, ultrasound investigation of the kidneys, urinary bladder and prostate with determination of residual urine, urodynamic examination of the lower urinary tracts. All the patients were divided into four groups. Group 1 of 10 patients received antiparkinsonian therapy which corrected voiding disorders. Group 2 (n = 26) received tolterodin in a dose 4 mg/day, group 3 (n = 14) received oxibutinin in a dose 5 mg/day, group 4 (n = 14) were treated with trospium chloride in a dose 30 mg/day. Thus, initial correction of antiparkinsonian therapy was made in 64 patients, after this correction voiding disorders reduced in 10 patients and no further treatment was needed. Voiding disorders reduced in all 3 groups of patients given the drugs. Cystometric capacity of the bladder increased in all 4 groups from 155 (86-450) ml to 220 (105-530) ml, on the average, and unstable contractions of the urinary bladder decreased from 3.1 (1-8) to 1.6 (0-5). Quality of life in the examinees improved considerably. The data obtained evidence for M-cholino-blockers efficacy in PD patients with unstable detrusor after correction of antiparkinsonean therapy. Two month course of tolterodin, oxibutinin and trospium chloride significantly reduced symptoms of urination disorders. PMID- 16708588 TI - [Diagnosis of infravesical obstruction in patients with benign prostatic hyperplasia]. AB - To improve reliability of the diagnosis of infravesical obstruction (IVO) in patients with benign prostatic hyperplasia (BPH), we examined 80 BPH patients (mean age 58.2 +/- 2.1 years). The examination included evaluation of complaints by IPSS, ultrasound investigation with determination of prostatic size and residual urine, and urodynamic tests: uroflowmetry, miction cystometry. We came to the conclusion that symptoms of BPH, size of the gland, amount of residual urine and maximal volumetric miction velocity not always evidence for IVO. To raise reliability of IVO diagnosis we propose the following formula: D1 = 0.818 x S(max) +0.0006 x S(min) +0.215 x Q(max) - 0.478 x Q(aver), where D1 is a discriminant function; S(max) is a maximal linear size of the prostate; S(min) is a minimal linear size of the prostate; Q(max) is maximal volumetric flow rate; Q(aver) is mean volumetric flow rate. If D1 > or = 2.85, IVO is definite. If D1 < or = 2.85, IVO absence is more probable. Thus, the proposed formula provides more reliable diagnosis of IVO in BPH patients than assessment of clinical indices. PMID- 16708589 TI - [Current combat wounds of the scrotum and its organs]. AB - Wounds of the scrotum and its organs are frequent among injuries of the urogenital system in local military conflicts. The scrotum is often wounded in mine explosions which result in severe multitraumas, crushing and abruptions. As a rule, scrotal wounds are not life threatening, but late diagnosis and inadequate surgery may provoke serious complications. Surgical policy should be most sparing and secure preservation of the scrotal organs. PMID- 16708590 TI - [Anaerobic parasitocenoses of the urinary tracts as a risk factor of acute pyelonephritis]. AB - We examined 86 patients with acute gestational pyelonephritis, 72 patients with acute pyelonephritis admitted to hospital, 63 control women with favourable obstetric anamnesis and physiological pregnancy which finished in birth of a healthy child and 57 women with favourable obstetric anamnesis undergoing pregravidal examination. Microbiological factors of acute pyelonephritis risk and acute gestation pyelonephritis were determined. These factors were the presence of anaerobic parasitocenosis in the urinary tracts represented with trichomonades, mycoplasms, ureaplasms, herpes viruses and cytomegaly in various combinations. PMID- 16708591 TI - [Efficacy of levofloxacin in the treatment of recurrent cystitis in women]. PMID- 16708592 TI - [Polyoxidonium in the treatment of calculous pyelonephritis]. AB - Efficacy of polyoxidonium was studied in combined therapy of urolithiasis complicated with secondary pyelonephritis. Of 60 patients with secondary pyelonephritis in urolithiasis, 30 received adjuvant polyoxidonium. Phospholipids and cholesterol in cytomembranes were defined with thin-layer chromatography. Humoral immunity was evaluated by the levels of IgA, IgM, IgG, hemolytic activity of the compliment system, the level of circulating immune complexes, activity of lysozyme. Concentrations of IL-1beta, TNF, IL-4, IL-6 were also estimated. Subpopulations of immunocompetent cells were tested on flow cytofluorimeter by immunofluorescence with monoclonal antibodies. Phagocytic component of the immune system was assessed by phagocytic activity of neutrophils, phagocytic count, NBT test. The trial demonstrates that polyoxidonium has a positive action on the disease course, an antiinflammatory, immunomodulating properties, suppresses peroxidation, promotes recovery of structural-functional characteristics of cellular membranes in patients with urolithiasis and secondary pyelonephritis. Polyoxidonium is recommended in secondary pyelonephritis and urolithiasis as adjuvant to basic therapy for anti-inflammatory, detoxication and immunomodulating effect. PMID- 16708593 TI - [Basic diagnostic algorithm in chronic pelvic pain syndrome]. AB - A basic diagnostic algorithm is proposed to improve efficiency of diagnosis of chronic inflammatory prostatitis. It is the sequence of the most necessary methods of examination which helps to clear a category of disease according to classification NIH (1995). A total of 57 patients with symptoms of prostatitis and quantity of leukocytes in the prostatic secretions < 15 per high power field. Use of this diagnostic algorithm allowed diagnosis of category IIIA in 36.8% patients free of inflammatory changes in the first expressed prostatic secretions and category IIIB in 8.8% patients with leukocytic count > 10, but < 15 per high power field. Thus, use of basic diagnostic algorithm in diagnosis of chronic prostatitis changed a category of chronic prostatitis in 45.6% cases among patients examined conventionally. PMID- 16708594 TI - [Reproductive function in chronic abacterial prostatitis patients]. AB - The results of examination of 325 patients with chronic abacterial prostatitis, varicocele showed marked changes in ejaculate parameters of these patients. Prostatitis of different duration in the same degree causes pathozoospermia, reduces ejaculate fertility. Such condition of spermatogenesis indicates involvement of other sex organs suggested by the literature data and previous experience. This necessitates extended diagnostic examination of patients with chronic prostatitis. PMID- 16708595 TI - [Vitaprost efficacy in patients with chronic abacterial prostatitis]. AB - We studied efficacy of a new cytomedine drug--vitaprost--in 98 patients with chronic abacterial prostatitis. Vitaprost was found to relieve symptoms of the disease, improved blood flow in the prostate, normalized blood rheology and immunological indices, increased mobility of spermatozoa in ejaculate. PMID- 16708596 TI - [Transrectal magnetotherapy of the prostate from Intramag device in prophylaxis of postoperative complications of transurethral resection of prostatic adenoma]. AB - The authors analyse preoperative preparation of 91 patients with benign prostatic hyperplasia (BPH). Two groups of patients received conventional preparation (group 1) and magnetotherapy (group 2) before TUR of the prostate. The examination covered immune system, bacteriological indices of urine and prostatic tissue. Infection of the urinary tract is a main risk factor of complications after TUR. Conventional preoperative preparation fails to correct immunity, to change bacterial urine flora, to improve hemodynamics in the prostate. Transrectal magnetotherapy with running magnetic field eliminates deficiency of T and B-cell immunity, raises functional activity of B-lymphocytes and phagocytic ability of neutrophils, reduces endogenic intoxication, tissue edema, bacterial contamination, number of thrombohemorrhagic complications. This leads to a decrease in the number of postoperative complications. PMID- 16708597 TI - [Fallourethroplasty with a free forearm flap in traumatic loss of the penis]. PMID- 16708598 TI - [Ectopy of the accessory ureter ostium]. PMID- 16708599 TI - [Migration of the intrauterine contraceptive in the urinary bladder with formation of the secondary stone and vesicouterine fistula]. PMID- 16708600 TI - [A giant ureterolith]. PMID- 16708601 TI - [Surgical treatment of Peyronie's disease]. PMID- 16708603 TI - Core measure compliance gets better with CPOE. PMID- 16708602 TI - JCAHO will want evidence that physicians are involved in safety. PMID- 16708604 TI - JCAHO looking closely at medication reconciliation. PMID- 16708605 TI - JCAHO award for stroke measures compliance. PMID- 16708606 TI - New NPSGs for 2007 will be big challenge. PMID- 16708607 TI - Manage staff fatigue to improve patient safety. Part 2 of 2. PMID- 16708608 TI - [Medico-legal and victimology aspects of homicides in the material of Bialystok Institute of Forensic Medicine in the years 1982-2003]. AB - The authors presented an analysis of homicides based on autopsy material collected at Institute of Forensic Medicine in Bialystok in the years 1982-2003. Of 8,780 postmortem examinations, this type of death accounted for 7,4 % of cases, including 31,2 % women and 68,8 % men. Married individuals predominated among females, whereas the majority of male victims were single. Fifty-seven percent of victims died on the crime scene, 5,3 % within 48 hours. In conflicts of long-standing, immediate conflicts and fights, the perpetrators most commonly employed hard, blunt or blunt-edged crime weapons. In cases of murder with robbery and sexual homicides, strangulation was the most frequently used method. No seasonal character in homicide prevalence was observed. PMID- 16708609 TI - [The role of ethanol in complex poisonings with carbon monoxide and hydrogen cyanide in fire victims]. AB - A total of 230 cases of deaths in burning spaces dating from the years 1995-2003 were investigated in Forensic Medicine Department, Silesian University of Medicine, Katowice. HbCO and HCN found in 177 blood samples ranged from 4-95 % (mean, 31,5 %) and 0,5-40,3 microg/ml (mean, 9,98 microg/ml), respectively. Moreover, ethanol was found in 122 blood samples. Its concentration ranged from 0,89-5,0 per thousand (mean, 1,45 per thousand). A comparative analysis of HbCO and HCN levels in the groups with and without ethanol showed that the range and the mean concentration of both these xenobiotics were higher in the group with no alcohol. It was also shown that the increased ethanol caused a drop in HbCO and HCN levels. To evaluate HbCO and HCN levels, the regression and correlation analysis was used. PMID- 16708610 TI - [DNAStat, version 1.0--a software package for processing a genetic profile database and for biostatistical calculations]. AB - The application of DNA studies to the administration of justice has led to the necessity of developing appropriate computer programs. Such programs must address two critical problems, i.e. the broadly understood data processing and archivization, and biostatistical calculations. This paper discusses DNAStat 1.0, a program that enables its user to 1) create and process an individual database; 2) analyze biological evidence by calculating the unconditional f and conditional p(X/X) profile frequency, with the possibility of taking into account the inbreeding (coancestry) coefficient, as well as setting the minimum allele frequency; 3) analyze paternity cases by calculating the paternity index PI and probability of paternity W for full and motherless trios, with the possibility of taking into account the silent allele frequency and prior probability. PMID- 16708611 TI - [Estimating the efficiency of DNA isolation methods in semen, blood and saliva stains using the QuantiBlot system]. AB - The aim of this study was to compare and select the optimal method of DNA isolation from blood, semen and saliva stains, as well as to determine appropriate conditions for employing amplification kits for identification of individual persons [brak w polskim tekscie]. The materials analyzed in this study consisted of stains of blood, semen and saliva samples stored for a year, and stains of blood stored for a month. Seven various methods of isolation were compared: the Fast DNA kit (Qbiogene), phenol/chloroform extraction, Sherlock (DNA II Gdansk), Dneasy (Qiagen), Wizard Genomic Purification Kit (Promega), Chelex 100 (Biorad) and salting out proteins method. After the isolation, the quantity of DNA was measured with QuantiBlot [brak w polskim tekscie]. The highest DNA concentration in bloodstains stored for one year and one month was observed employing the salting out proteins method. The phenol-chloroform extraction method was also found to produce reasonably good results. Isolation from blood and semen with salting-out method appeared to be the most effective. The phenol/chloroform method was dependent on the age and origin of the materials [brak w polskim tekscie]. The Sherlock kit was proven to be effective in blood samples stored for one year. DNA concentration values obtained in semen and saliva samples were very low and characterized by a low repeatability. PMID- 16708612 TI - [Medicolegal investigation system in the state of Connecticut]. AB - The paper explains the organization of medicolegal investigation system conducted in the State of Connecticut and regulated by the law of that state. The author presents issues connected with reportable deaths, possible types of medicolegal investigations, people responsible for deciding whether autopsy should be performed, individuals responsible for conducting autopsy, and other responsibilities of medical examiners in association with medicolegal investigations. The paper does not provide a detailed description of the organizational structure and legal basis of the medical examiner system in Connecticut due to space limitations. PMID- 16708613 TI - [Methods of preliminary evaluation of psychophysical skills of drivers]. AB - Drug and medication use-associated impairment of driving skills is increasingly becoming a serious problem. Detecting such DUI drivers is very difficult due to the diversity of drugs that affect the central nervous system (CNS), or to the possibility of using several drugs simultaneously. In practice there are numerous different simple tests, which do not require special equipment and can be used for preliminary detection of persons, who are suspected of taking drugs causing impairment of driving skills. In order to popularize the knowledge on the subject of testing drivers for drug use, the authors review the most commonly used methods of driving skill impairment evaluation reported in the literature, and present the scope of employing such tests in various countries. PMID- 16708615 TI - [Depression and legal capacity to stand trial]. AB - The criteria for assessment of legal capacity to stand trial have long been a significant issue in the Polish criminal law. The main problem in opinionating practice is the fact that the code of penal procedure and the executive penal code do not provide any univocal criteria of a mental disease that should be met according to the legal regulations in force. Because of their nature and lack of uniformity, depression and affective disorders pose a particularly great problem for experts who should opinionate on the legal capacity of the suspected and the accused to stand trial. On the one hand, it cannot be accepted that a mental illness renders a person unable to substantially understand legal proceedings and to make a rational defense. On the other hand, however, emotional reactions and mild-degree depression disorders that are only natural when an individual violates law should not be allowed to be regarded as a grave disease and to paralyze legal proceedings. In the present study, the authors have attempted to describe the guidelines that should be followed by court-appointed experts in psychiatry that are commissioned by the court to assess the mental state of an individual. The report emphasizes that in such cases, a thorough analysis, based on medical and legal premises and the experience of the involved court experts are necessary. PMID- 16708614 TI - [Murder as fun]. AB - Aggressive behaviors are often accompanied by excitement. Violence is often triggered to as a means of helping a felon reach an emotional thrill. This is also true in the case of homicides. Hedonistic motivation of a murderer derives from a need to feel an emotional 'high' from killing a human being. In such instances, a murderer's pattern of behavior reflects elements of fun and entertainment. PMID- 16708616 TI - [Dog-bite related deaths]. AB - The author presents two cases of fatal dog bites. In both, the casualties were small children: a newborn and a two-month infant. In the first case, the injuries were inflicted by a puppy, in the second, by three blood-related dogs kept at home. PMID- 16708617 TI - Unusual form of accidental mechanical asphyxia. AB - This brief case report shows an unusual form of accidental atypical strangulation. A 46-year-old male became trapped between the rims of an automatically closing door during his hopeless effort to enter a supermarket at night. The victim's neck was violently pressed by the edges of the automatic door. From external observation we found an incomplete ligature mark and numerous small abrasions in the neck. On autopsy we found typical signs of asphyxia and fractures of the hyoid-laryngeal complex and bruising of the soft tissues of the neck. The subsequent toxicology was negative with the exception of alcohol level. We came to the common conclusion of a double form of mechanical asphyxia - atypical violent strangulation and atypical hanging. The third highly likely contributory factor of death was direct stimulation of the baroreceptor nerve endings in the left carotid sinus and left carotid sheath. In recent literature, there have been presented only a few cases describing this manner of asphyxia and death. PMID- 16708618 TI - [Myocarditis in two cases of HIV infection]. AB - The authors present two cases of death in young persons infected with HIV virus and diagnosed with inflammatory changes of the heart muscle rarely seen in forensic medical practice. In one of the cases, only the findings revealed upon autopsy did prompt the examiner to suspect the infection, even though before death, the patient had been in several hospitals. PMID- 16708619 TI - Most EDs 'fail' key tests in mock drills for pediatric trauma cases. AB - Make sure your staff and your facility are well prepared to handle pediatric trauma patients. Emphasize the differences between children and adults, i.e., estimating proper weight to determine fluid and meds requirements; smaller body surface and its implications for blunt trauma victims; and the need for warmer room temperatures. Make sure they are well schooled in the use of intraosseous needles. Conduct drills on a regular basis to improve team readiness and ensure appropriate staffing and proper placement of equipment. PMID- 16708620 TI - ED docs at high risk for suicide. AB - Make your staff aware of the resources that are available to them, and create an atmosphere in which they will feel more comfortable to seek help: State physicians' health programs and employee assistance programs offer confidential services. Hold regular staff inservices to create a dialogue about stressful situations and to discuss signs and symptoms of depression. Approach staff members who appear troubled and let them know you are there to help them. PMID- 16708621 TI - Doctor in triage slices door-to-discharge times. AB - Eliminating time-consuming processes up front and taking a hard look at how you treat nonemergent patients can yield dramatic results: One ED cut pain-to-first meds in half, cut 30 minutes off door-to-discharge times, and reduced left before being seen rates by about one-third. Having a doc in triage can help speed and ensure appropriateness of advance ordering. If your fast-track process is not meeting its turnaround goals, consider other alternatives such as Mid Care. PMID- 16708622 TI - First EDs to undergo regular unannounced surveys warn: be prepared for questions. AB - ED managers undergoing the first regular unannounced accreditation surveys report that surveyors are heavily interested in the ED. Be sure you are notified and notify your staff as soon as surveyors arrive. Use a last-minute checklist. (See p. 3) Expect patients to be traced to the ED and for your staff to be questioned. Be prepared for surveyors to emphasize National Patient Safety Goals, including those on read-back of orders/test results and hand off communication. Interdepartmental teams and quizzes can help your staff be continuously prepared. PMID- 16708623 TI - Systemwide collaborative targets capacity woes. AB - Adopting engineering principles of flow and collaborating with other EDs can lead to the adoption of new and effective strategies for improving your ED's ability to handle capacity challenges. One hospital system achieved a 33% improvement in the number of admissions from the ED completed in fewer than 90 minutes. Innovative tools such as "Top 10 Ways to Make Throughput Worse" result from sharing of ideas. Intranet site and regular meetings can help all participants share challenges as well as successes. PMID- 16708624 TI - 'Flow techs' help ED run smoothly. AB - Creating a new position of flow technicians (flow techs) can relieve physicians of a wide range of responsibilities, while making life easier for nurses and other staff members as well. Flowtechs can handle discharge instructions, locate paperwork, and ensure orders have been followed. Nurses no longer have to wait in line to request information from the ED physician. Having a human interface improves communication among physicians, patients, nurses, and families. PMID- 16708625 TI - [Etiopathogenesis of atherosclerosis--a clinical problem that is still relevant]. AB - Atherosclerosis is a complex dynamic pathological problem. Apart from classic risk factors of atherosclerosis development (age, gender, arterial hypertension, elevated cholesterol concentration particularly LDL fraction, cigarette smoking, diabetes, hyperhomocysteinemia, low physical activity, obesity, mental stress), infectious-inflammatory factors play an important role in the onset of atherosclerotic changes. Vascular endothelium is the main target organ of their activity. Basing on recent literature the study presents pathomechanisms of the development of endothelial dysfunction caused by the risk factors of atherosclerosis, particularly by infectious-inflammatory factors and their mutual interactions which lead to the development of stable and unstable atheromatous plaque. PMID- 16708626 TI - [CD28 and CTLA-4 costimulatory molecules expression on T lymphocytes and NK cells in nonallergic bronchial asthma]. AB - The aim of the study was to assess the role of costimulatory molecules in pathogenesis of nonallergic bronchial asthma. MATERIAL: The studied group consisted of 30 patients with nonallergic asthma, 24 with nonallergic asthma and recurrent respiratory tract infections and 20 healthy controls. RESULTS: In nonstimulated and lipopolysaccharide (LPS) stimulated cultures of control group the mean value of CD16+ + 56+ CD28+ subpopulation were significantly decreased compared to values in both types of asthma. Similar dependence was observed in cultures of asthmatic patients with infection and healthy person, stimulated with IL-15 and in CD4+CD28+, CD8 +CD28+ and NKCD28+ cultures from all studied groups, stimulated with LPS plus IL-15. In nonallergic asthma and infection asthma mean values of CD4 +CD152+ and CD16+ +CD56 +CD152+ from nonstimulated and IL-15 stimulated cultures were significantly increased compared to analogical values in control group. CONCLUSION: The results of our studies suggest that in nonallergic asthma the disturbed expression of costimulatory molecules may play an important role in pathomechanism of disease. PMID- 16708627 TI - [Influence of treatment of inhaled corticosteroids on the function of the larynx in asthmatic patients]. AB - The objective of the study was to examine influence of inhaled glycocorticosteroids on phoniatric function of the larynx in patients suffering from asthma. MATERIAL AND METHODS: Thirty minutes after the administration and long-term therapy effect of the inhaled glycocorticosteroids on local condition of the phoniatric organ was assessed in 15 patients with recently diagnosed bronchial asthma. The diagnostics of asthma included: subjective and objective examination, prick tests, spirometry, total IgE and histamine test for bronchial hyperactivity. Evaluation of voice function was implemented subjectively and objectively with use of videolaryngostroboscopy (VLSS). Acoustic assessment of voice was also performed. RESULTS: Administration of glycocorticosteroids as inhaled discs resulted in incidents of cough, mouth and throat dryness, sensation of polydipsia and skin inflammation around the mouth. Occasionally, hoarseness and discomfort in the larynx area were noted. On long-term administration, dysphonia, hoarseness and voice fatigue due to dysfunction of innermost larynx muscles, particularly vocal fold adductors. After 30 minutes of the glycocorticosteroid administration in patients with recently diagnosed asthma, irritation of the pharynx and larynx mucosa was recorded. CONCLUSION: The long term treatment with inhaled glycocorticosteroids resulted in myopathy of proper muscles of the larynx. PMID- 16708628 TI - [Self-medication with antibiotics in Poland]. AB - Antibiotic resistance, the important public health threat, depends on antibiotic overuse/misuse. Self-medication with antibiotics is of serious medical concern. The aim of the study, as a part of SAR project (Self-medication with antibiotic in Europe) was to survey the incidence of this phenomenon. MATERIAL AND METHODS: In Poland among 938 respondents of the questionnaire 27% adults were treated with antibiotics at least once a year, 13% of the therapies were undertaken without medical consultation. The most frequently prescribed antibiotic was amoxicillin, the most often self-ordered--fusafungine. RESULTS: The main self-medicated symptoms were sore throat and common cold. The duration of self-ordered therapy was less then 5 days. The sources of antibiotics used for self-medication were leftovers and those given by friends or family. CONCLUSION: The results of the study confirmed that antibiotic self-medication is relatively frequent problem in Poland and interventions are required in order to reduce the frequency of antibiotics misuse. PMID- 16708629 TI - [Antibiotics use for acute gastroenteritis in ambulatory care of children before and after implementation the healthcare system reform in Poland]. AB - AIM OF THE STUDY: Our study aimed to investigate the ambulatory antibiotics use for acute gastroenteritis in children admitted to the hospital in 1998-2000. MATERIAL AND METHODS: The retrospective analysis of medical records was performed in 369 children up to two-years of age; including 87 admitted in 1998, 136 - in 1999 and 146 in 2000 year. Results. Antimicrobials use decreased year after year. In 1998 the antibiotics therapy was included in 44.9% of children, in 1999--in 35.3% and in 2000 year in 32.2% (p = 0.02). In 1998 year, the antibiotics were ordered mainly for children diagnosed as bacterial diarrhea (76.9%) compared to viral etiology (18%). In 2000 year the antibiotics were prescribed for viral gastroenteritis twice more often (61.7%) than for bacterial etiology (29.8). Children with acute gastroenteritis and coexistent signs and symptoms were given antibiotics for treatment diarrhea in the similar percentage as those without any: in 1998--54.2 and 45.8%, in 1999--50.8 and 49.2%, in 2000--40.3 and 59.7%, respectively. Microbiologic examination was not performed in any case before treatment. CONCLUSIONS: This study demonstrates the discrepancies between recommendations and ambulatory practice in the antibiotic treatment of acute diarrhea in children both before and after implementation the healthcare system reform. Education programs are needed for optimal management of acute gastroenteritis in children including training in necessity of coproculture before prescribing antimicrobials. PMID- 16708630 TI - [The concentration of plasma anion oxalate in children treatment antibiotics beta lactame]. AB - The aim of work was the assessment of plasma anion oxalate (Ox) concentration in children during antibacterial treatment depending on way and time of antibiotic administration. MATERIAL AND METHODS: The examinations were carried out in 80 children, without nephrolithiasis, aged 10.1 +/- 4.3 years with bronchopneumonia, treated with beta-lactame antibiotics. The children were divided in two groups: I -children treated with oral amoxicillin + clavulanic acid or cefuroxime axetil (n=40), II--children treated with the same antibiotics intravenously (n=40). The Ox concentration in plasma and urine was measured using an enzymatic method with oxalate oxidase, four times. (0)--before treatment, (a)--in third day and (b)--in last day of administration (10 to 14 day), (c)--3 weeks after finishing treatment with antibiotics. RESULTS: The result showed that in children before treatment (0) mean plasma Ox concentration was 2.439 +/- 0.645 micromol/l. In 3rd day (a) the Ox concentration increased to 7.848 +/- 0.999 micromol/l (p < 0.01), in last day of treatment (b) decreased to 5.681 +/- 0.871 micromol/l, and after 3 weeks (c) came back to initial values (p > 0.05). Intravenous antibiotics administration did not influence plasma Ox concentration. CONCLUSIONS: Plasma oxalate concentration increases during oral administration of beta-lactame antibiotics caused by increased intestinal absorption, as a result of saprophytic microflora deterioration. However intravewous administration of the same antibiotics does not change the concentration of plasma oxalate. PMID- 16708631 TI - [Thyroglobulin concentration after introduction of population iodine prophylaxis in random selected group of children and adolescents in Szczecin region]. AB - The analysis from the early 1990s showed the iodine deficiency and endemic goitre in whole country. These studies contributed to introduce in 1997 the obligatory model of iodization the table salt. The risk connected with iodine deficiency causes that the monitoring of iodine balance became a very important thing. One of the most important indicators is thyreoglobulin serum concentration. Aim of the study was determination of thyreoglobulin serum concentration as an assessment the efficacy of current model of iodine prophylaxis in random selected group of children and adolescents in Szczecin region. MATERIAL AND METHODS: The study was undertaken in the years 2003-2004. In random selected group of children and adolescents in Szczecin region. The patients administered vitamins or minerals enriched with iodine were excluded from a study. The study included 151 persons aged 9 to 16 years, 80 boys (53%) and 71 girls (47%). Radioimmunometric and radioimmunological methods were used for the determination of thyreoglobulin (Tg) and thyrotropin (TSH) and free thyroxine (FTJ concentrations. Iodine excretion with urine was determined on the basis of catalytic Sandell-Kolthoff reaction. Collected data were then analyzed using sophisticated computer-based statistic program. RESULTS: In whole study group mean Tg concentration was 11.97 +/- 7.57 ng/ml. In 69 persons (45%) they were normal, in 82 (55%) were increased: 67 (44%) of them has insignificant increase (low iodine deficiency) and in 15 (11%) were in a range 20-40 ng/ml, what is connected with a mean iodine deficiency. In whole study group mean FT4 concentration was 15.59 +/- 4.72 pmol/l. In a group with normal thyreoglobulin concentrations mean FT4 was 17.69 +/- 4.53 pmol/l and was significantly higher than in a group with insignificant and mean Tg increase and were 15.36 +/- 4.43 pmol/l and 13.72 +/- 4.63 pmol/l, respectively. Mean TSH concentration was 2.23 +/- 0.9 microU/ml and mean iodine concentration was 87.2 +/- 37.5 microg/l. Between TSH serum concentrations and ioduria were no significant differences between the groups. There was significant correlation between thyreoglobulin concentration and FT4 (r(s) = -0.24) and thyrotropin (r(s) = 0.21). No correlation was found between concentration of these hormones and ioduria. Similarly, there was no correlation between thyreoglobulin concentration and iodine urinary excretion. CONCLUSIONS: Thyreoglobulin serum concentrations evaluated during the study revealed iodine deficiency in Szczecin region. Correlations between thyreoglobulin concentrations, free thyroxine and thyrotropin make a Tg determination to be more helpful indicator of iodine balance than ioduria. PMID- 16708632 TI - [Influence of gastric juice pH on the metabolism of vitamin C in gastric mucosa and juice]. AB - Vitamin C is present as ascorbic acid (Asc) with antioxidative properties and as its oxidation product dehydroascorbic acid (dAsc). Asc is actively transported from blood to gastric juice where it prevents formation of carcinogenic nitrosamines and protects mucosa from reactive oxygen species. Another important function of gastric mucosa is proton pump-dependent secretion of hydrochloric acid. The pump maintains high pH gradient between gastric juice and plasma. The aim of the work was to analyse relations between gastric juice pH and concentrations of Asc and dAsc in plasma, gastric mucosa and juice as well as other factors modifying metabolism of vitamin C and function of gastric mucosa. MATERIALS AND METHODS: 31 patients were subjected to diagnostic endoscopy due to dyspepsia. Concentrations of Asc and dAsc were measured in plasma, gastric mucosa and juice with HPLC method. pH of gastric juice was determined. Histopathology examination of mucosa and urease test for Helicobacter pylori were performed. Data concerning dyspeptic symptoms and used drugs, including vitamin C preparations, were collected. RESULTS: Patients taking vitamin C preparations had significantly lower gastric juice pH values than the others (median 2.2 vs 5.4; p < 0.01). Treatment with vitamin C preparations was also associated with higher Asc concentrations in gastric juice (median 16.6 vs. 1.8 micromol/L; p < 0.09); the difference was statistically significant in the subgroup of patients with gastritis (median 16.6 vs. 0.1 micromol/L; p < 0.04). Strong negative correlations of pH with Asc (Rs = -0.67; p < 0.001) and dAsc (Rs = -0.48; p < 0.01) concentrations were observed in gastric juice. Positive correlation of pH and relative dAsc content expressed as percent of total vitamin C concentration in gastric juice was also statistically significant (Rs = +0.48; p < 0.05). CONCLUSIONS: Performance of the proton pump and vitamin C metabolism in the stomach are closely mutually connected. Modification of hydrochloric acid secretion may affect gastric juice vitamin C concentration. Potentially disadvantageous influence of antisecretory drugs on the mechanisms of antioxidative protection in the stomach needs further investigation. PMID- 16708633 TI - [Whole blood citrulline levels in patients with coeliac disease]. AB - Citrulline is a non-protein amino acid synthesized by the intestine. Whole blood citrulline concentrations in patients with coeliac disease have not been reported. The aim of the study was determination of the amino acid concentrations in coeliacs on gluten-free diet and gluten-containing diet. MATERIAL AND METHODS: 61 patients with coeliac disease were studied. Whole blood citrulline were determined in dried blood spots by tandem mass spectrometry. One-way analysis of variance and the Bonferroni post hoc test for multiple comparisons were used to determine differences among groups under study. RESULTS: Mean citrulline levels were higher in patients on strict gluten-free diet comparing to those newly diagnosed (32.2 +/- 8.7 vs. 24.9 +/- 5.7 micromol/l; p = 0.025). CONCLUSION: Whole blood citrulline concentration may be a simple biochemical marker for diagnosis and monitoring enterocyte loss in coeliac disease. PMID- 16708634 TI - [The assessment of the influence of natural coffee and its modified form on the level of homocysteine, vitamin B6 and folic acid in healthy volunteers]. AB - The aim of our investigation was the assessment of the influence of natural coffee and that modified by water and pressure extraction (60% less of 2-methyl isoborneol) on the level of homocysteine, folic acid and vitamin B6 in healthy volunteers. MATERIAL AND METHODS: The study was conducted on 36 healthy volunteers. 20 women and 16 men; smokers constituted half of the group. The study was conducted as a double blind trial (coffee without labels) after randomization into two groups. Initially for 4 weeks, one group drank natural coffee and the other a modified one. After four weeks there was a 28-day break in drinking coffee, after which the groups swapped roles and another trial lasted for the subsequent 4 weeks. All people examined drank three servings of coffee a day brewed from 13 g of material in 180 ml of boiling water. Throughout the entire experiment the examined subjects did not change their diets and did not take any vitamin supplements. Blood for analysis was drawn four times and the following analyses were carried out. homocysteine, folic acid, vitamin B6 total, LDL and HDL-cholesterol, triglicerides, Lp(a) and fibrinogen. RESULTS: We found a significant increase level of plasma homocysteine from 9.6 to 11.4 micromol/l (p < 0.001) in persons drinking natural, unfiltered coffee. However drinking modified coffee free from irritants resulted in a tendency towards lowering the level of homocysteine (from 9.1 to 8.7 micromol/l). CONCLUSIONS: From the above study it may be concluded that lowering the content of irritants in coffee results in reducing its undesired influence on the homocysteine level. Reduction of natural coffee consumption or its change on coffee with lowering the content of irritants should be recommended to cardiovascular disease persons. PMID- 16708635 TI - [Anti-inflammatory action of myoinositol in renal insufficiency]. AB - AIM OF THE STUDY: Release of prostaglandin E2 (PGE2) and leukotrien B4 (LTB4) in vitro by resting and PHA-stimulated peripheral blood mononuclear cells (PBMNC) in the presence of three concentrations of myoinositol (30, 300, 600 micromol/l) was investigated. MATERIAL AND METHODS: We examinated 10 uremic patients on regular hemodialysis treatment and 10 healthy subjects (control group). RESULTS: Release of PGE2 and LTB4 by resting and PHA-stimulated PBMNC was significantly lower in the presence of myoinositol in concentrations generally obserwed in the blood serum of chronic uraemic patients on regular hemodialysis treatment (600 micromol/l) in both investigated groups, while it remained unchanged in the presence of myoinositol in the concentration observed in normal blood serum (30 micromol/l). CONCLUSION: The results seem to indicate that myoinositol, in the concentrations found in uremic blood serum, may possibly exert antiinflammatory actions. PMID- 16708636 TI - [Ocular and orbital symptoms in benign sinonasal neopasmas]. AB - Osteoma (0.01-0.4%) and inverted papilloma (0.5-4%) are the most common benign tumours of the nose and paranasal sinuses, whereas heamangioma occurs very rare. Orbital and ocular symptoms are uncommon in these types of neoplasmas. AIM OF THE STUDY: Analysis of symtoms, diagnostic procedures and treatment options in 10 patients with diagnosed orbital and ocular complications of benign tumours of the nose and paranasal sinuses. MATERIAL AND METHODS: There are 4 cases of inverted papilloma, 3 cases of osteoma and 3 cases haemangioma. All patients had laryngological, ophtalmological examination, radiografic evaluation by plain film -4 cases, CT--9 patients and angiography in 1 case. Presenting orbital and ocular symptoms occured in 7 patients and in 3 left person orbital involvement was presented only radiographically and intraoperatively. In 5 cases there were correlation between orbital symptoms and radiologic findings. The most common presenting feature--orbital pain (6 patients) and proptosis (5 patients) were observed in papilloma and haemangioma. In 8 cases there were signs of bone destruction in CT, what was intraoperatively confirmed in 4 cases. RESULTS: Nine of the patients were operated: 4--lateral rhinotomies, 2--Denker's operations, 2- operations of sinus frontalis and/or ethmoidalis by external approach and in 1 case resection of maxilllary sinus's tumor were performed. In 2 patients with inverted papilloma there were recurrences 3 years followed operation. The rest of patients stay without recurrences of tumor. CONCLUSION: Orbital and ocular symptoms in benign tumors of nose and paranasal sinuses do not always correlate with radiologic and intraoperative findings. PMID- 16708637 TI - [Personality and coping with stress in the general practitioner and surgeon's profession]. AB - Personality variables of the professional are an essential determinant of the stress experienced by a professional and the active steps taken to cope with or counter those problems. AIM OF THE STUDY: To verify this hypothesis, an analysis of the relationship between personality variables of specialists of the various fields and their individual patterns in dealing with professional demands was conducted. MATERIAL AND METHODS: 30 each of general practitioners and surgeons constituted the study material, whose personality traits levels and patterns of coping behaviour and work-related attitudes (AVEM) were measured. RESULTS: Comparing the results obtained in the two groups of specialists, a significant differentiation in the levels of their personality traits, as well as within patterns applicable when dealing with professional demands and strains (p < .001). Among surgeons risk pattern A (The 'Excessively-ambitious' pattern), whereas among general practitioners risk pattern B (The Resigned/burnout' pattern). CONCLUSIONS: The results of the study confirm the postulate that allows for personality variables in research in coping behaviour patterns (AVEM) and in prophylactic or remedial programmes created for concrete professional groups. PMID- 16708638 TI - [Thermographic assessment of thermal effects of Er:YAG laser in periodontal surgery]. AB - AIM OF THE STUDY: An assessment of thermal effect of Er:YAG laser (KEYII, KaVo) on oral soft tissues in select procedures. MATERIAL AND METHODS: Experimental researches were carried out on Wistar rats. To measure the temperature changes the thermal imaging camera (ThermaCAM SC3000, FLIR Systems) was used. RESULTS: There has been a significant increase of temperature observed on the end of optical fibre: the mean temperature ranged from 270 to 360 degrees C (at laser energy of 100 mJ and repetition rate of 25 Hz) and from 230 to 290 degrees C (300 mJ, 15 Hz). On the surface of oral mucosa thermal changes at the time of laser frenulectomy was analysed along the line of incision. The temperature above 50 degrees C was recorded on the length of 2 mm (at 100 mJ) and 3 mm (at 300 mJ). The temperature maintained on this level for about 0.4 s. On the surface of tongue during lingual mucosa excision the temperature above 40 degrees C was observed on the length of 1.6 mm (80 mJ, 2 Hz) or 2.5 mm (160 mJ, 2 Hz). The rate of cooling for both cases was lower than 0.5 s. CONCLUSION: To prevent undesirable thermal side effects from an Er:YAG laser optical fibre should be moved very fluently in non-contact mode. PMID- 16708639 TI - [Expression of chemokine receptor CCR5 in patients with type 2 diabetes]. AB - Overexpression of chemokine receptors might contribute to the initiation of atherosclerosis, due to promotion of monocyte migration to the subendothelial space which is considered as an initiating mechanism in the natural history of atherosclerosis. The CCR5 receptor is one of the chemokine receptors present on peripheral T lymphocytes, monocytes and macrophages. OBJECTIVE: The aim of the study was to determine the expression of CCR5 receptor on peripheral blood mononuclear cells (PBMC) in patients with type 2 diabetes. MATERIAL AND METHODS: 14 patients with type 2 diabetes and 6 healthy subjects as a control were included to the study. We evaluated the density of the CCR5 receptor on peripheral blood mononuclear cells by flow cytometric analysis by use of LSR Cytofluorograf System (Becton Dickinson Co.). RESULTS: There was significantly higher density of the CCR5 receptor on PMBC in diabetic subjects when compared to the controls. CONCLUSIONS: (1) Patients with type 2 diabetes present overexpression of the CCR5 receptor on peripheral blood mononuclear cells. (2). Overexpression of the CCR5 receptor should be considered as a factor that promotes the development of atherosclerosis in diabetic patients. PMID- 16708640 TI - [Predisposing factors and results of pharmalogical treatment using ursodeoxycholic acid of gallbladder stones in children]. AB - Gallbladder stones (GBS) in adults is one of the most often disease of the gastrointestinal tract, it occurs in about 0.2% of children. Risk factors of the GBS in children are different that those in adults. Therapeutical standards in GBS depend on the age, clinical symptoms, complications and coexistent diseases. AIM: The estimation and comparison of risk factors of GBS in children in two periods (1989-1996 and 1997-2004), and the estimation of the results of the pharmacological treatment with UDCA. MATERIAL AND METHODS: In 112 children aged from 3 months to 18 yrs. with GBS recognized in the period of 1989-2004 the estimation and comparison of the frequency of risk factors in two periods: 1989 1996 (39 children aged from 4 months to 17 yrs.) and in years 1997-2004 (73 children aged from 3 months to 18 yrs.) was done. In the second step we analyzed results of pharmacological treatment with UDCA in 41 children after completion of the therapy. Only children with noncalcifying stones smaller than <15 mm in diameter were qualified to the treatment. Statistical analysis (chi2-test) was used to estimate results of the analysis. RESULTS: Gallbladder stones was recognized more often in girls (65%), obesity was present in 42% of children, family history of the GBS was positive in 28.6%, prior severe disease (e.g. sepsis) in 15.9%. In 16% of children GBS was idiopathic. In the period of 1997 2004 significantly more often than in the period of 1989-1996 in children with GBS were observed: serum lipid abnormalities (28.4 vs. 10.3%; p < 0.03), prior severe diseases requiring intensive care (21.6 vs. 5.1%; p = 0.02) and intensive reducing of the body weight (9.5 vs. 0%; p < 0.05). Any of the risk factors had influence on the positive result of pharmacological treatment. Dissolution of the small stones (<5 mm) in comparison with stones bigger than 10 mm (p < 0.05) was observed significantly more often. CONCLUSIONS: (1) Gallbladder stones in children was observed more often in girls, children with prior severe disease, and in obese children with the positive family history of GBS. (2) Good results of pharmacological treatment of GBS in children depends on the size of the stones. (3) Risk factors of the GBS have no significant influence to the positive result of the pharmacological treatment using UDCA in children. PMID- 16708641 TI - [In vitro modification of antioxidant response and its influence on cytokine synthesis in children with renal failure]. AB - Recently it was shown that oxidative stress has a negative influence on immunological response. The aim of the study was to investigate the influence of N-acetylcysteine on intracellular oxidative stress and cytokine synthesis in peripheral blood lymphocytes in children with end stage renal disease (ESRD). MATERIAL AND METHODS. We examined 25 children (age 2-16 ys.) with ESRD. To determine oxidative stress we used dihydrorhodamine 123 (DHR). Intracellular oxidation of DHR in T. lymphocytes reflected intracellular oxidative stress within cells. The intracellular synthesis of cytokines (IL-2, IFN-r, IL-4, IL-6) was detected with flow cytometty RESULTS. We found lower intracellular stress and higher cytokines synthesis after incubation with N-acetylcysteine (NA C) in all subpopulations of lymphocytes. CONCLUSIONS. Our results shows that patients with ESRD present increased oxidative stress in T lymphocytes which may lead to decreased cytokines synthesis and abnormal immune response. NAC has favorable influence on these abnormalities. PMID- 16708642 TI - [Latent coeliac disease in a 4.5-year-old girl with type 1 diabetes mellitus- case report]. AB - Four and half year-old girl with type I diabetes mellitus and latent coeliac disease was presented. The typical serological screening at type I diabetes mellitus onset was negative. Three years later serological screening was performed again because of recurrent diarrhoea. At the time serum IgA antiendomysial antibody was positive. The intestinal biopsy showed subtotal villous atrophy with intraepithelial lymphocyte infiltration and elongated crypts. It seems that diabetic children should be periodically checked by screening tests for coeliac disease. PMID- 16708643 TI - [BRAF gene mutation in thyroid cancer]. AB - Mutations of genes coding effectors of signaling pathway RET/PTC-RAS-RAF-MEK-ERK, involved in cell growth and proliferation, are important in papillary thyroid cancer development. To earlier discovered mutations of RAS and RET/PTC genes, BRAF gene mutation has been recently added. Mutation of BRAF gene appears in various types of carcinomas, but most frequently in malignant melanomas (66%) and papillary thyroid cancer (average 44%). The BRAF gene protein product belongs to the serine-threonine kinase family and to the RAF proteins subfamily, among which it is the strongest activator of MAP kinases cascade. The most frequently mutation of BRAF gene is thymine to adenine transversion at nucleotide position 1796 (T1796A). This point mutation causes valine to glutamic acid substitution at residue 599 (V599E), that results in constitutive and oncogenic activation of BRAF kinase. The relation between mutations of BRAF, RAS and RET/PTC genes has not been found, although they together exist in two thirds of papillary thyroid cancers. BRAF(TI796A) oncogene appears in papillary thyroid cancer, whereas it has not been found in follicular thyroid cancer and benign thyroid adenomas. For this reason mutated BRAF gene could be specific molecular marker, with relatively high sensitivity in diagnostics of papillary thyroid cancer. In addition, BRAF gene has been demonstrated as a novel prognostic biomarker, which correlates with unfavorable clinicopathological factors, such as extrathyroidal invasion and distant metastasis. PMID- 16708644 TI - [Thyroid hormone resistance syndrome]. AB - Resistance to thyroid hormones (RTH) is an inherited syndrome characterised by reduced target tissue responsiveness to these hormones. In the recent years, it has become clear that RTH is probably much more common than is generally recognised, and is often misdiagnosed and inaccurately treated. Subjects suffering from RTH have raised serum thyroid hormone levels and raised or inappropriately normal thyrotropin levels. Two major forms of a clinical presentation of this disorder are asymptomatic or slightly symptomatic subjects with generalised resistance and patients with thyrotoxic features suggesting predominant pituitary resistance. Surprisingly, these various clinical situations are determined by the same genetic defect. In this paper, aetiology, symptoms, clinical classification, diagnosis and treatment of RTH are reviewed with putting special emphasis on the results of recently published studies. PMID- 16708646 TI - [Extracellular matrix metalloproteinases. Part II]. AB - Remodeling of extracellular matrix is possible due to activity of specific, skilled proteolytic enzymes present in connective tissue. Metalloproteinases (MMPs) are involved most of physiological and pathological processes in human body. To date 28 enzymes have been identified and 23 in humans. This paper is a systemic review of metalloproteinases family. Identification of enzymes in ethiopathogenesis of various diseases is useful in modern therapy planning. PMID- 16708645 TI - [Matrix metalloproteinases. Part I]. AB - Metalloproteinases (MMPs) are the extracellular matrix enzymes indispensable in physiological and pathological matrix remodeling. They are produced intracellulary as endopeptidases and secreted to the extracellular space. Under normal conditions MMPs are present in tissues at low levels which increases dramatically during any changes. To date 28 enzymes have been identified and 23 in humans. This paper is a review of metalloproteinases structure, their natural and artificial inhibitors and regulation mechanisms. Function and distribution are critical for pathogenesis and therapy of diseases. PMID- 16708647 TI - [Matrix metalloproteinases and tissue inhibitors of metalloproteinases in the pathogenesis of rheumatoid arthritis]. AB - Rheumatoid arthritis (RA) is a chronic inflammatory disorder of joints characterized by the accumulation of mononuclear cells and the proliferation of the synovium-lining layer. The role of lymphocytes, macrophages and fibroblasts infiltrating the synovium is not fully understood. These cells are supposed to be involved in the tissue destruction by several mechanisms, including the production of proinflammatory cytokines and matrix metalloproteinases. Matrix metalloproteinases (MMPs) are the enzymes that participate in the proteolytic degradation and remodelling of the extracellular matrix. Their action is controlled by tissue inhibitors of metalloproteinases (TIMPs). In this review we describe the role of metalloproteinases and their tissue inhibitors in the pathogenesis of rheumatoid arthritis. PMID- 16708648 TI - [Hypercortisolism in pregnancy]. AB - Hipercortisolemia in pregnancy occurs rarely because of menstrual disturbances and infertility in subjects with Cushing syndrome. The diagnosis of pathological hipercorisolemia in pregnant women is often difficult because some symptoms of the disease may be associated with complicated pregnancy. Hipercortisolemia causes weight gain, especially trunk obesity with thin extremities, arterial hypertension, glucose intolerance, purple-blue skin striae and hirsutism. Hormonal and radiological diagnostic in pregnancy is limited. Results of hormonal estimations and of dynamic tests are difficult to interpret because of physiological changes in the hypothalamo-pituitary-adrenal axis connected with pregnancy. Interpretation of magnetic resonance imaging of pituitary gland should include hypertrophy of pituitary gland connected with pregnancy. The choice of optimal time and method of treatment should be cautious and individualized because of the possibility of maternal and fetal complications. PMID- 16708649 TI - [Congenital adrenal hyperplasia due to steroid 21-hydroxylase deficiency]. AB - Congenital adrenal hyperplasia is a general term applied to a group of several inherited enzymatic defects of cortisol biosynthesis. The most frequent cause of this disease is by far 21-hydroxylase deficiency which is considered one of the commonest metabolic disorders. The degree to which the activity of this enzyme is diminished correlates with the severity of congenital adrenal hyperplasia and therefore the clinical presentation of 21-hydroxylase deficiency has a wide spectrum of clinical and laboratory abnomalities. The recent developments have improved prenatal diagnosis and treatment of affected females to minimise genital virilisation. Despite progress made in its recognition and treatment, diagnosis and management of 21-hydroxylase deficiency is still the subject of many debates and controversies. In this paper, aetiology, symptoms, diagnosis and treatment of 21-hydroxylase deficiency in various groups are reviewed with putting special emphasis on the results of recently published studies. PMID- 16708650 TI - [Vascular endothelial growth factor in head and neck cancer]. AB - Angiogenesis plays a major role in tumor progression and metastasis. The family of vascular endothelial growth factors (VEGF) and their receptors with tyrosine kinase activity are main mediators of angiogenesis. Some researchers interested in angiogenesis assessed a value of determination of VEGF and his receptors in tissues and serum of patients with head and neck cancer. The correlation between clinical and pathological parameters and the level of the proangiogenic factor, their prognostic value and the influence on tumor radioresistance was evaluated. PMID- 16708652 TI - [Standard operating procedures in ethic committees]. AB - Polish ethic committees should have to work together in order to maintain and develop high quality standards in the protection of human subjects. Exchanging knowledge, know-how and information polish ethic committees should have to implement standard operating procedures. Procedures should improve quality and proficiency of all types of ethic committee's activities. Standard operating procedures should cover as important problems as conflict of interest, trial's insurance or elections of ethic committees. The opinions of experts who have been reviewing medical research projects for several years may prove to be especially valuable in this setting. Governmental initiatives and creation of forum for polish ethic committees are essential in the effective standardisation, coordination and implementation of procedures in regional ethic committees. These projects need support via public funding from our authorities. PMID- 16708651 TI - [Coercion in the treatment of anorexia nervosa]. AB - Anorexia nervosa (AN) is a complex disorder of unclear aetiology, nowadays one of the most serious chronic diseases of adolescent girls and young women. Severe weight loss and associated behaviours may endanger anorexic patients' lives. A characteristic for the disorder the reluctance in accepting treatment can lead in such situations to the necessity of involuntary treatment. The authors have presented indications for hospitalization as well as have discussed consequences, short- and longterm effectiveness of coercion measures in therapy of anorexia. The necessity of further investigations on this problem has been stresssed, particulary connected with good prognosis in the involuntary treatment. The lack of one-valued legal bases for involuntary treatment of anorexia in polish legislation has been mentioned as well as the lack of proper medical outposts providing them complex treatment. PMID- 16708653 TI - [Sepsis--actual clinical problem]. AB - An incidence of sepsis, septic shock, systemic inflammatory response syndrome (SIRS) and multiple organ dysfunction syndrome (MODS) is still actual clinical problem. Inducing factors and clinical pictures are similar to recently reported data from comparable populations in North America, Europe and Australia. The most important complication of severe sepsis is organ dysfunction observed in over 30% of sepsis patients hospitalized in intensive care unit. Applied intensive therapy including new generations of antibiotics gives an increase in clinical recovery. However, a hospital mortality of sepsis patients is over 30%. PMID- 16708654 TI - [The role of the maxillary incisors in the development of the base of the nose. Applications in dento-facial orthopedics]. AB - The neonatal respiratory distress observed in the event of a solitary median maxillary central incisor compels us to reconsider some of the traditional concepts relative to the transverse growth of the nasal level of the face. The "container-contents" connections associating maxillary incisor odontogenesis with the development of the premaxillary and facial envelopes draw the attention to the significant geometrical and mechanical expression of this morphogenesis. They require attributing to the maxillary incisors an important motor role in this development. They lead to granting the ontogenetic bonds, between malocclusions and disturbed nasal breathing, the place they deserve, taking into account the morphological integration combining them. They eventually open a new therapeutic prospect: the optimization of the development of the growing face, with regard to the various tissue mechanics and physiologies, becoming the best guarantor for the prevention of relapse after dentofacial orthopedics. PMID- 16708655 TI - [Growth of the skull and bony kinetics interfering with facial morphogenesis. Conceptual bases of success in orthopedic treatments before the age of 6]. AB - In order to perform an orthopedic treatment without relapse, one needs to identify properly the cranial patterns responsible for the malocclusion and act on them before the age of 6. PMID- 16708656 TI - [Genetics and the dento-facial complex]. AB - Recently mutations in single genes have been found to cause dentofacial deviations like hypodontia, cleft lip and palate, as well as some craniofacial malformation syndromes like Aperts/Crouzon syndromes, cleidocranial dysostosis... These findings may lead to an overestimation and oversimplification of the role of genes in our daily life. The "one gene, one disease" principle has been applied too often, while most complex human characteristics are determined by a large number of genes, each leading to an increased liability for a certain condition. Whether this condition is expressed or not depends upon the environment the individual is exposed to as well as upon the position of a certain gene in the genetic network. The question is not whether a condition is genetically or environmentally determined, but to what extent environment and genes play a role and how they interact with each other. As early as 1870 Galton with his classical twin studies initiated the Nature-Nurture debate, whose immediate result was a great belief in the primacy of "Nature". Later, after Darwin proposed his theory of evolution (1871), the pendulum swung too far in the direction of "Nurture". At present, the dominance of the Nature camp has returned because of exceptional genetic scientific progress. Again new insights are expected that will clarify the mutual interaction of genes and between genes and the environment. After deciphering DNA, scientists need to carry out more research to explain how it is translated into the human phenotype, in health and in disease. Therefore, analysing the phenotypic variance with detailed clinical data--especially in twins--will contribute equally as much as DNA analysis to elucidating the functioning of our genes. More than their concordance, the discordance of monozygotic twins will be extremely instructive for the task of determining the genotype-phenotype relationship as well as for explaining the role of epigenetic phenomena. PMID- 16708657 TI - [Cranial asymmetries and their dento-facial and occlusal effects]. AB - Cases of occlusal asymmetry detected in the temporary dentition are frequently associated with skeletal asymmetries of the cranial base. They should be corrected as soon as possible. Symmetry of these dental groups is achieved with the use of mini-screws implanted in designated sectors of the hard palate. When a new occlusal plane derived from the corrected maxilla is obtained, the mandible will re-center itself in relation to it. As a result cranial bones can remodel and the possibility of induced temporo-mandibular disorders is greatly reduced. PMID- 16708658 TI - [Influence of feeding method on maxillofacial development]. AB - The functional differences between breast-feeding and bottle-feeding may have a significant effect on dento-facial development and on the genesis of some severe malocclusions. Orthodontists should be aware of this sometimes under-estimated aspect of development. They should, in conformity with their role as professional health care providers, advise long-term breast feeding as a way of preventing certain disorders or, at least, reducing their severity. PMID- 16708659 TI - [Tongue disfunction and abnormal development]. AB - Almost all the disorders seen in the mixed and adult dentitions were already present in the temporary dentition. We should ask ourselves what postural and dynamic role the tongue plays in defective development of the dental arches. PMID- 16708660 TI - [Interception in the primary dentition: mastication and neuro-occlusal rehabilitation]. AB - After evoking the characteristics of masticatory movements, their control and their adaptability, a review of the literature illustrates how the masticatory work and its variations can influence the dentoalveolar, as well as the maxillary growth, by adapting the morphological structures to the functional context. The conceptions of Planas, and especially his "neuro-occlusal rehabilitation", are widely explained: unilateral alternate mastication, "Planas' Functional Masticatory Angles (PFMA) and the vertical dimension, the "atrophies" of maxillary development resulting from reduced mastication, the "dominant unilateral mastication syndrome" and, of course, all the original treatments introduced by Planas. These specific orthodontic treatments can be applied as interceptive procedure as early as during the deciduous dentition. The selective occlusal grinding, implemented to reduce and balance the "PFMA", and the expansion devices using "running track plates" are described in concepts and clinical uses. Planas' Class II treatments of Class II, also using "direct occlusal bonded tracks" and several mandibular propulsion auxiliaries are explained as well as the early treatments of lateral cross-bite and the use of the "Equiplan". Mastication, the first of the functions assigned and carried out by the masticatory apparatus, is a genuine "functional matrix" able to stimulate the growth of the dental arches and of the maxillaries. When perfectly understood and applied, the "neuro-occlusal rehabilitation" according to Planas allows to restore a functional balance for the masticatory apparatus from an early age, and subsequently, reorientates growth to a morphological normalization. PMID- 16708661 TI - [Class II interceptive therapy in the mixed dentition: intra-oral distalization appliances]. AB - Treatment of Class II malocclusion during the mixed dentition may sometimes require upper molar distalization. To achieve this, many devices have been suggested. The purpose of this article is to classify them and to point out their effects on the dentition and upon skeletal structures. PMID- 16708662 TI - [Orthopedic activators for growth and treatment of Class II malocclusion]. AB - Activators are functional jaw orthopedic appliances which provide various mandibular position, but always different from habitual positions. Mechanisms and modus operandi of activators explain therapeutic objectives of clinicians. When we analyse different determinants of modus operandi of activators (skeletal, muscular and articular), it is possible to classify these appliances in rigid monobloc appliances, composite or elastic activators and thrusting activators. Many prospective or retrospective clinical studies analyse dentoalveolar or skeletal effects of the activators. PMID- 16708663 TI - [Optimal time for treatment with the Herbst appliance]. AB - This article endeavours to answer the question of whether treatment at an early or a later age is the ideal time for Herbst therapy. We discuss this topic by summarizing and evaluating the short and long-term effects of the Herbst appliance on the dentofacial complex. We place special emphasis on treatment outcomes in relation to the patient's somatic and skeletal maturity at the time of therapy. We also address the long-term stability of the treatment results. The available knowledge in the literature demonstrates that the ideal period for Herbst treatment is in the permanent dentition at or just after the peak of pubertal growth. PMID- 16708664 TI - Poaching ban needed to save Africa. PMID- 16708665 TI - Safe on the home watch. AB - For some children with heart conditions anticoagulation is mandatory. A pioneering, nurse-led service allows families to monitor blood at home, saving them repeated hospital visits. Home monitoring requires careful family selection and training. Warfarin doses are adjusted by the hospital team when families phone in their results. PMID- 16708666 TI - Promoting evidence-based practice in stroke care in Australia. AB - AIM: To explore approaches to the promotion of evidence-based practice from academic and clinical perspectives by visiting acute stroke units and collaborating centres of the Joanna Briggs Institute, an international network of academic centres. METHOD: A semi-structured interview schedule was developed, piloted and used to guide interviews with academic and clinical staff in five state capital cities in Australia. Data were analysed and findings reviewed by clinical and academic participants. FINDINGS: Four distinct but not mutually exclusive models and common but variously applied pathways for translation of evidence into clinical practice were identified. Key influential factors included context and local culture, the nature of evidence and role of clinical expertise. Implementation and change management strategies were recognised as emerging priorities. CONCLUSION: A range of methods to advance research synthesis, dissemination and knowledge transfer into clinical practice were demonstrated and may warrant consideration for the UK. PMID- 16708667 TI - The role of student ambassadors in university recruitment. AB - Recruitment to nursing programmes is becoming increasingly difficult. At Staffordshire University we applied to a project called Higher Education Full Circle for the funding of paid student ambassadors to help with our nurse recruitment programmes. Two nursing students took on the role while continuing with their training. This article discusses the work of the student ambassadors. PMID- 16708669 TI - Clinical governance. PMID- 16708668 TI - Malaria in UK travellers: assessment, prevention and treatment. AB - Malaria is the most serious tropical disease. Increasing numbers of people are travelling to tropical destinations where they are at risk of malaria. Nurses need to be aware of the disease risk, prevention of mosquito bites and appropriate chemoprophylaxis to protect the health of travellers. This article describes the malaria lifecycle, bite prevention, chemoprophylaxis, diagnosis and prevention strategies for people travelling to malarious areas. Additional resources are supplied for nurses who want further information. PMID- 16708670 TI - Getting personal. PMID- 16708671 TI - Adventure and altruism. PMID- 16708672 TI - The bump sum. PMID- 16708673 TI - iPLEDGE: protecting patients or prohibiting access to care? PMID- 16708675 TI - Acne scarring: current treatment options. AB - Acne vulgaris is one of the most common skin disorders treated in dermatology. Subsequent scarring that may result from acne can lead to devastating consequences. There are currently a wide variety of treatment modalities available for acne scarring. The most widely used options are reviewed. PMID- 16708674 TI - Allergic contact dermatitis to preservatives. AB - In summary, a wide variety of skin care products contain preservatives. Patients who are allergic to one of these preservatives may have either localized or widespread dermatitis. Affected patients may find it difficult to avoid thimerosal without the help of the health care provider because the use of these allergens is so widespread. Patch testing is an invaluable tool for patients who struggle with dermatitis. Antigen-avoidance lists that facilitate patient education about what products to avoid are available from the manufacturers of patch test allergens (for example, TRUE Test or Chemotechnique). These lists are helpful starting points for patients in that they provide general categories (for example, shampoos, soaps, or creams) of products that the patient should avoid. With these printed guidelines alone, patients must read skin care product labels carefully, looking for the names of their allergens as identified by patch tests as well as for any synonyms and cross-reactors of these allergens. Thus, patients may feel overwhelmed by hearing the names of allergens that are long and complex. After an allergen has been identified, the nurse can play a key role in helping patients understand their dermatitis and its management. Nurses are in a unique position to spend time educating patients about how to uncover the sources of specific allergens and, subsequently, how to avoid them. The Contact Allergen Replacement Database can help in this educational process by giving patients a shopping list of specific items that are free of the specific allergens causing their allergic contact dermatitis. PMID- 16708676 TI - What's your assessment? Basal cell carcinoma. PMID- 16708677 TI - Assessing phytophotodermatitis: boy with erythema and blisters on both hands. AB - Phytophotodermatitis (PPD) is a phototoxic reaction which occurs when the skin comes in contact with a photosensitizer and is subsequently exposed to radiation. PPD is often seen in people handling furocoumarin-containing products, such as agricultural workers, bartenders, florists, and gardeners. It may also be seen in beachgoers, athletes, and children. The pattern of the lesions usually resembles streaks; the hands and mouth are most commonly affected due to eating and handling of the offending furocoumarin-containing agents. PMID- 16708678 TI - Skin breakdown in acute care pediatrics. AB - The purpose of this study was to develop a simple, single-page measurement tool that evaluates risk of skin breakdown in the peadiatric population and apply it to the acutely hospitalized child. Data were collected over a 15-month period from 347 patients on four in-patient units (PICU, medical-surgical, oncology, and adolescents) on skin breakdown using the AHCPR staging guidelines and compared to the total score on the Starkid SkinScale in order to determine its ability to predict skin breakdown. The inter-rater reliability of the Starkid Skin Scale was r2 = 0.85 with an internal reliablity of 0.71. The sensitivity of the total score was low (17.5%) but highly specific (98.5%). The prevalence of skin breakdown in the acutely hospitalized child was 23%, the majority (77.5%) occurring as erythema of the skin. Buttocks, perineum, and occiput were the most common locations of breakdown. Occiput breakdown was more common in critically ill (PICU) patients while diaper dermatitis was more common in the general medical surgical population. PMID- 16708679 TI - Neuropathic ulcers: diabetes and wounds, part II. Differential diagnosis and treatment. PMID- 16708680 TI - Connective tissue nevus. PMID- 16708681 TI - Tinea versicolor. PMID- 16708683 TI - How to recruit, interview, and retain employees. AB - Decide how you will recruit new employees and place your advertisements. Screen the resumes and select the applicants to interview. Prepare yourself for the interview by reviewing the job description and determining the qualities you are seeking and the questions required to gain the information you seek. Conduct the interview and followup. Select the final candidate, extend an offer, and establish a start date. Don't forget to notify unsuccessful candidates. Once the employee starts, orient her to the practice and set up a training schedule. Monitor her progress and develop a system of ongoing evaluation of her performance. Make sure you take the necessary steps to retain employees. Remember, hiring and firing employees is an expensive endeavor. PMID- 16708682 TI - Skin rashes in a patient with antibodies to Ro/SS-A. PMID- 16708684 TI - The keratinocyte. PMID- 16708685 TI - Cutaneous malignancies: BCC, SCC, and MM. PMID- 16708686 TI - The Johnny Appleseeds of oranizational change. AB - Change cannot be executed by merely announcing a new program. Just as Johnny Appleseed carried and planted apple seeds that sprouted throughout the Northwest Territory, there are seed carriers in every organization who are carrying the seeds of positive change or a negative reaction to change. Performative ties can carry the advantages of message integrity, reducing the cost of knowledge transfer, and rapid transfer of knowledge. The future of leadership needs to move from focusing on extraordinary individuals to extraordinary combinations of people that will create extraordinary collaboration. PMID- 16708687 TI - Influence of hospice use on hospital inpatient mortality: a state-level analysis. AB - This study tests the hypothesis that high hospice enrollment is associated with lower Medicare inpatient mortality. The results show that Medicare inpatient mortality in a state can be explained by hospice enrollment and a host of demographic and market environment variables. An increase in hospice population by 100 individuals is associated with a reduction of 28 inpatient deaths, ceteris paribus. The results suggest, among other things, that opportunities exist for greater expansion of hospice capacity in low-use states to reduce deaths in the expensive hospital setting and improve the quality of end-of-life care for terminally ill patients. PMID- 16708688 TI - The applicability of Bass's model of transformational, transactional, and laissez faire leadership in the hospital administrative environment. AB - The purpose of this study is to evaluate empirically in the hospital administrative environment the relationship of leadership behaviors to subordinate manager's perceived outcomes, through examination of B. M. Bass's (1985) model of transformational, transactional, and laissez-faire leadership. The author measured leadership orientation and outcome factors through subordinate managers' ratings of hospital CEOs using a questionnaire, which asked: Is there a relationship between the leadership styles of hospital CEOs and subordinate managers' self-reported willingness to exert extra effort, perception of leader effectiveness and satisfaction with their leader? Findings revealed that the relationship between transformational leadership and the outcome factors were stronger and more positive than were the transactional and laissez-faire styles. These findings are consistent with the hierarchal patterns reported and support the universality of the model. PMID- 16708689 TI - Does money really matter? A review of the literature on the relationships between healthcare organization finances and quality. AB - The authors review empirical literature from 1980 to 2005 on relationships between healthcare organizations' finances and quality of care. They found only 16 studies of this topic that employed statistical methods. This research indicates cumulatively that expenses, fiscal margin, and asset and liability management all affect healthcare outcome quality. There is less evidence about how organizational finance factors affect structural or process quality, and there is no information about how structural or process quality mediates between finances and outcomes. The authors note what patterns have emerged from previous studies and make specific suggestions about what future research is necessary and why. PMID- 16708690 TI - Katrina: lessons from the aftermath. PMID- 16708691 TI - Employee evaluation strategies for healthcare organizations--a general guide. PMID- 16708692 TI - Agricultural chemical safety assessment: A multisector approach to the modernization of human safety requirements. AB - Better understanding of toxicological mechanisms, enhanced testing capabilities, and demands for more sophisticated data for safety and health risk assessment have generated international interest in improving the current testing paradigm for agricultural chemicals. To address this need, the ILSI Health and Environmental Sciences Institute convened a large and diverse group of international experts to develop a credible and viable testing approach that includes scientifically appropriate studies that are necessary without being redundant, and that emphasize toxicological endpoints and exposure durations that are relevant for risk assessment. Benefits of the proposed approach include improved data for risk assessment, greater efficiency, use of fewer animals, and better use of resources. From the outset of this endeavor, it was unanimously agreed that a tiered approach should be designed to incorporate existing knowledge on the chemistry, toxicology, and actual human exposure scenarios of the compound, with integration of studies on metabolism/kinetics, life stages, and systemic toxicities. Three international task forces were charged with designing study types and endpoints on metabolism/kinetics, life stages, and systemic toxicities to be used in the tiered approach. This tiered testing proposal departs from the current standardized list of hazard studies used by many national authorities, and represents the first comprehensive effort of its kind to scientifically redesign the testing framework for agricultural chemicals. PMID- 16708693 TI - The acquisition and application of absorption, distribution, metabolism, and excretion (ADME) data in agricultural chemical safety assessments. AB - A proposal has been developed by the Agricultural Chemical Safety Assessment (ACSA) Technical Committee of the ILSI Health and Environmental Sciences Institute (HESI) for an improved approach to assessing the safety of crop protection chemicals. The goal is to ensure that studies are scientifically appropriate and necessary without being redundant, and that tests emphasize toxicological endpoints and exposure durations that are relevant for risk assessment. Incorporation of pharmacokinetic studies describing absorption, distribution, metabolism, and excretion is an essential tool for improving the design and interpretation of toxicity studies and their application for safety assessment. A tiered approach is described in which basic pharmacokinetic studies, similar to those for pharmaceuticals, are conducted for regulatory submission. Subsequent tiers provide additional information in an iterative manner, depending on pharmacokinetic properties, toxicity study results, and the intended uses of the compound. PMID- 16708694 TI - A tiered approach to systemic toxicity testing for agricultural chemical safety assessment. AB - A proposal has been developed by the Agricultural Chemical Safety Assessment (ACSA) Technical Committee of the ILSI Health and Environmental Sciences Institute (HESI) for an improved approach to assessing the safety of crop protection chemicals. The goal is to ensure that studies are scientifically appropriate and necessary without being redundant, and that tests emphasize toxicological endpoints and exposure durations that are relevant for risk assessment. The ACSA Systemic Toxicity Task Force proposes an approach to systemic toxicity testing as one part of the overall assessment of a compound's potential to cause adverse effects on health. The approach is designed to provide more relevant data for deriving reference doses for shorter time periods of human exposure, and includes fewer studies for deriving longer term reference doses that is, neither a 12-month dog study nor a mouse carcinogenicity study is recommended. All available data, including toxicokinetics and metabolism data and life stages information, are taken into account. The proposed tiered testing approach has the potential to provide new risk assessment information for shorter human exposure durations while reducing the number of animals used and without compromising the sensitivity of the determination of longer term reference doses. PMID- 16708695 TI - A tiered approach to life stages testing for agricultural chemical safety assessment. AB - A proposal has been developed by the Agricultural Chemical Safety Assessment (ACSA) Technical Committee of the ILSI Health and Environmental Sciences Institute (HESI) for an improved approach to assessing the safety of crop protection chemicals. The goal is to ensure that studies are scientifically appropriate and necessary without being redundant, and that tests emphasize toxicological endpoints and exposure durations that are relevant for risk assessment. The ACSA Life Stages Task Force proposes a tiered approach to toxicity testing that assesses a compound's potential to cause adverse effects on reproduction, and that assesses the nature and severity of effects during development and adolescence, with consideration of the sensitivity of the elderly. While incorporating many features from current guideline studies, the proposed approach includes a novel rat reproduction and developmental study with enhanced endpoints and a rabbit development study. All available data, including toxicokinetics, ADME data, and systemic toxicity information, are considered in the design and interpretation of studies. Compared to existing testing strategies, the proposed approach uses fewer animals, provides information on the young animal, and includes an estimation of human exposure potential for making decisions about the extent of testing required. PMID- 16708696 TI - Cancer nursing--1986 or 2006? PMID- 16708697 TI - The role of the data safety monitoring board: why was the Avastin phase III clinical trial stopped? AB - In early February 2006, Genentech Inc. issued a joint press release with Roche Holding Corporation explaining that their phase III early-stage colon cancer trial of Avastin (bevacizumab, Genentech Inc., South San Francisco, CA) would be stopped after the deaths of four patients. This announcement was the result of preliminary review of the data by the Data Safety Monitoring Board (DSMB) that was formed to monitor the study. What is the DSMB? What criteria does it use to determine whether a study should be stopped early? The purpose of this article is to (a) explain the role of the DSMB, (b) give an overview of the Avastin Adjuvant (AVANT) study, and (c) provide information and resources about clinical trials for practicing RNs in oncology. PMID- 16708698 TI - Abdominal complaints: diverticular disease. AB - CASE PRESENTATION: A patient with a history of follicular large cell lymphoma schedules an urgent office visit for vague complaints of intermittent abdominal pain for the past month. The pain is present in the lower quadrants and is associated with cramping and alternating periods of constipation and diarrhea. The patient denies nausea, vomiting, or fevers. A review of systems is negative for any other complaints, and the patient's weight is stable. The patient is concerned that the pain represents a recurrence of lymphoma, which originally was diagnosed eight years prior. The patient was treated for relapse more than two years ago with no evidence of disease recurrence. A computed tomography (CT) scan completed five months prior was negative for lymphadenopathy. A previously enlarged right inguinal node had resolved completely. However, mild diverticulosis was observed in the sigmoid colon. The following information represents a general review of diverticular disease. PMID- 16708699 TI - Assessment and pharmacotherapy of depression. AB - Depressive disorders are experienced by a significant number of patients with cancer, with reported rates of 0%-58% (Massie, 2004). Numerous studies have demonstrated that depression in patients with cancer is underdiagnosed and inadequately treated as a result of a number of factors (Schwartz, Lander, & Chochinov, 2002). Although more severe or complex situations involving depression or other mental illness necessitate specialist referral, sometimes professionals treating patients with cancer may find the need to initiate treatment. Thus, clinicians need to be aware of basic principles related to the assessment and treatment of depression. PMID- 16708700 TI - Care of patients with neutropenia. AB - Neutropenia can be a significant problem in the oncology setting. Awareness of potential risks, management of neutropenia, and preventive measures guide nurses in providing comprehensive care that can make the difference between life and death. PMID- 16708701 TI - Cognitive impairment in patients with brain tumors: assessment and intervention in the clinic setting. AB - Cognitive impairment is the most common neurologic problem associated with brain tumors and is present in many people with brain tumors from the time of diagnosis. Treatment of primary brain tumors with surgery, radiation, chemotherapy, and adjunctive medications such as corticosteroids results in further adverse effects on cognitive function. To plan the best care for patients with brain tumors, healthcare providers must initiate systematic and accurate assessment of cognitive functioning at the first clinic visit and extend assessment throughout the course of illness. This article outlines the range of cognitive dysfunction that may be seen in patients with primary brain tumors and offers information for clinicians seeking to develop their skills and implement a systematic approach to cognitive screening. The use of cognitive screening to guide timely intervention, such as referral to a neuropsychologist and the provision of anticipatory guidance to people with brain tumors and their families, is discussed. PMID- 16708703 TI - Advances in the management of low- to intermediate-risk myelodysplastic syndrome: Integrating the National Comprehensive Cancer Network guidelines. AB - Myelodysplastic syndromes (MDSs) are a group of hematologic diseases that present unique challenges for oncology nurses, especially because patients with the disorders are being seen more often in oncology practices. An increasing array of therapeutic options are available, and the National Comprehensive Cancer Network published its first clinical practice guidelines for MDSs in 2004. This article provides oncology nurses with the most recent data on supportive care as well as emerging therapies for patients with low- to intermediate-risk MDS. PMID- 16708704 TI - Fertility considerations for female adolescent and young adult patients following cancer therapy: A guide for counseling patients and their families. AB - During the past 20 years, an increase in survival among pediatric patients with cancer has occurred, and a need exists to be proactive prior to cancer therapy regarding issues related to fertility. Chemotherapy and radiation can cause a number of deleterious side effects to female patients, including early menopause and its associated side effects of osteoporosis and heart disease as well as the inability to carry a pregnancy to term or conceive a child. Many drugs and fields of radiotherapy are associated with an increased incidence of female reproductive complications. Options are available for the preservation of female fertility, but many are experimental. Of highest importance is the need to counsel female adolescents and young adults before beginning induction chemotherapy or radiation. Nurses play a large part in the information about future fertility that female patients receive before the initiation of cancer therapy. After reading this article, nurses will have a better understanding of the impact of cancer therapy on the female reproductive system and be more comfortable discussing the topic with their patients. PMID- 16708705 TI - Ocular changes with oxaliplatin. AB - Ocular toxicity, although uncommon, can occur with many chemotherapeutic agents. Platinum compounds have been documented to produce a variety of ocular side effects, and reports have been made of ocular toxicity with oxaliplatin. This article reports on four patients who experienced ocular symptoms while receiving oxaliplatin. The symptoms included tunnel vision and visual loss with postural changes. One patient had objective findings that included papilledema. All of the changes were reversible. Oxaliplatin will continue to be used widely, so clinicians treating patients with it must be alert for unusual toxicities such as those described in this article. PMID- 16708706 TI - Hormone-refractory prostate cancer: a shifting paradigm in treatment. AB - Prostate cancer, the most common male cancer, affects one in eight American men. Risk factors for the disease include increased age, race, and family history of prostate cancer. To date, surgery, radiation, and hormonal therapy have been the mainstays of treatment. In the past, chemotherapy served only a palliative role for men with prostate cancer and failed to produce a survival advantage or any significant measurable disease response. However, for the first time, docetaxel based regimens have demonstrated improved survival in men with hormone-refractory prostate cancer in two different, large, phase III studies. Additionally, a number of novel agents are being developed with the hope that treatment for men with hormone-refractory prostate cancer will be improved. Oncology nurses provide critical symptom management strategies as well as education to men with prostate cancer and their partners. Therefore, maintaining current state of the knowledge about best practices and treatment for prostate cancer is crucial. This, in turn, directs efforts to educate patients and family members about treatments and management of side effects. PMID- 16708707 TI - Before the teaching begins: Managing patient anxiety prior to providing education. AB - Patients experiencing cancer also can experience anxiety. Moderate to severe levels of anxiety can interfere with patients' ability to concentrate and comprehend new information. The condition is particularly troublesome when trying to present educational material related to recommended treatment interventions. Patients' understanding of the material is critical to ensure informed consent. Informed consent can be compromised if patients are unable to understand the information being provided. Nurses must be cognizant of the impact that anxiety has on patient education and assess patients prior to initiating patient teaching. By managing anxiety before beginning education, nurses can provide an environment more conducive to learning. PMID- 16708708 TI - Preventing vincristine administration errors: Does evidence support minibag infusions? AB - Although vincristine sulfate is indicated for IV use only, it has been inadvertently administered intrathecally. Accidental vincristine administration via the spinal route (intrathecally via a lumbar puncture or intraventricularly via an Ommaya reservoir) causes rapid sensory and motor dysfunction, usually followed by encephalopathy, coma, and death (Schulmeister, 2004). Autopsy findings include grossly edematous and congested brain and spinal cord tissue, with axonal degeneration and myelin loss of the spinal nerves (Kwack et al., 1999; Williams et al., 1983). PMID- 16708709 TI - Oral mucositis. AB - CASE STUDY: Ms. J is a 55-year-old woman who was diagnosed with stage IIIA colon cancer three months ago. Prior to her diagnosis, she experienced an unintentional 17-pound weight loss. She underwent an exploratory laparotomy with colon resection. Following recovery from her surgery, she began an adjuvant chemotherapy regimen of 5-fluorouracil, leucovorin, and oxaliplatin. She presents to the clinic today for her third chemotherapy treatment. PMID- 16708710 TI - Surrogate markers are not 'one-size-fits-all'. PMID- 16708711 TI - ERCP: current uses and less-invasive options. AB - Endoscopic retrograde cholangiopancreatography (ERCP) is well suited for the evaluation and treatment of diseases of the bile ducts and pancreas, but it carries the risk of inducing pancreatitis. Magnetic resonance cholangiopancreatography (MRCP) and endoscopic ultrasonography have exceptional value in imaging the gallbladder, common hepatic duct, common bile duct, and pancreas. These imaging studies have replaced ERCP for diagnostic purposes in patients with a low pretest probability of finding lesions amenable to endoscopic therapy, such as bile duct stones. PMID- 16708712 TI - When do common symptoms indicate normal pressure hydrocephalus? AB - The Adams triad (gait impairment, dementia, and urinary incontinence) of normal pressure hydrocephalus (NPH) is well known, but other illnesses present with similar symptoms. Accurate diagnosis of this rare disorder depends on careful evaluation. A ventriculoperitoneal shunt is the only effective therapy, and deciding whether the patient will benefit is the final challenge in the evaluation process. PMID- 16708713 TI - A tropical souvenir not worth picking up. PMID- 16708714 TI - Heel pain: diagnosis and treatment, step by step. AB - The differential diagnosis of heel pain is broad and can be overwhelming if a systematic approach is not used. Focused questions and physical examination can help identify heel pain as Achilles tendinopathy or plantar fasciitis, or as due to a less common cause such as gout, spondyloarthropathy, or hypercholesterolemia. PMID- 16708715 TI - Diagnosing primary osteoporosis: it's more than a T score. AB - Although densitometry has contributed immensely to detecting primary osteoporosis, it is only a tool that generates some useful numbers to guide diagnosis. The T score, a leading diagnostic marker for primary osteoporosis, must be put in its proper context. It is but one measurement that is quite useful in one cohort of patients, namely, postmenopausal women older than 60, but it can be misleading in others. The z score is a more descriptive measurement of bone loss in younger patients. However, both the T score and z score are limited in their diagnostic potential and must be incorporated with other diagnostic aspects, such as family history, laboratory results, and genetic influences. In the end, physicians diagnose osteoporosis, not densitometry. PMID- 16708716 TI - Exenatide and pramlintide: new glucose-lowering agents for treating diabetes mellitus. AB - Insulin is not the only hormone that regulates plasma glucose levels. Glucagon like peptide 1 (GLP-1), produced in the small intestine, and amylin, produced by beta cells in the pancreas, also have glucose-lowering effects. Synthetic analogues of these hormones are now available for clinical use. PMID- 16708717 TI - Addressing the challenges of cardiorenal syndrome. AB - In heart failure, as the heart gets worse, often so do the kidneys, complicating the treatment of heart failure and worsening the prognosis. This article addresses challenges in the use of diuretics, angiotensin-converting enzyme (ACE) inhibitors, and other therapies in the cardiorenal syndrome, as well as novel therapies that hold promise, such as arginine vasopressin antagonists, adenosine A1 receptor antagonists, and ultrafiltration. PMID- 16708718 TI - Does the acidity of self-etching primers affect bond strength and surface morphology of enamel? AB - PURPOSE: This study examined the ultrastructure and microtensile bond strengths (TBS) of self-etching (with different acidity) and conventional adhesive systems bonded to unground enamel. MATERIALS AND METHODS: Resin composite (Filtek Z250) buildups were bonded to unground enamel surfaces of third molars after adhesive application with the following materials: Clearfil SE Bond (CSE); Optibond Solo Plus Self-Etch (OP); Tyrian Self Priming Etching (TY), and the controls Scotchbond Multi-Purpose Plus (SBMP) and Single Bond (SB). Six teeth were assigned to each material. After storage in waterfor 24 h at 37 degrees C, the bonded specimens were sectioned into beams of approximately 0.8 mm2 and subsequently subjected to microTBS testing at a crosshead speed of 0.5 mm/min. The average values were subjected to one-way ANOVA (alpha = 0.05). The effect of surface conditioning of each material was observed under scanning electron microscopy (SEM). RESULTS: The highest resin-enamel bond strength was observed for SBMP (22.7 +/- 5.2) and SB (26.7 +/- 5.2 MPa). The lowest mean bond strengths were 10.9 +/- 3.2 and 7.8 +/- 1.5 MPa for TY and OP, respectively. CSE showed an intermediate performance (18.7 +/- 4.6 MPa). An overall increase in porosity was evident along the entire enamel surface treated with the self-etching primers; however, no selective demineralization similar to that with 35% phosphoric acid was observed. CONCLUSION: The highest bond strength means and the more retentive etching pattern were observed for the two-step etch-and-rinse adhesives. Among the self-etching systems studied, Clearfil SE Bond should be preferred. PMID- 16708719 TI - Microtensile bond strengths of composite to dentin treated with desensitizer products. AB - PURPOSE: This study was designed to analyze the influence of desensitizing procedures on dentin bond strength. MATERIALS AND METHODS: Forty bovine incisors were used, divided into four groups (n = 10): G1: control; G2: Gluma Desensitizer (Heraeus Kulzer); G3: Oxa-Gel (Art-Dent); G4: low-intensity laser (MMOptics). The buccal surface was wet ground flat with 180-, 400- and 600-grit silicon carbide abrasive paper to expose midcoronal dentin and create a uniform surface. After the application of the desensitizing agents to the exposed dentin, the specimens were etched with 35% phosphoric acid for 30 s, and an adhesive (Single Bond) was applied and light cured. A 4-mm high crown of composite resin (Filtek Z250) was then built up. Specimens were trimmed to an hourglass shape with cross sections of 1 mm2. Each specimen was individually fractured by a microtensile testing machine at a crosshead speed of 0.5 mm/min. The data, recorded in MPa, were analyzed with one-way ANOVA and the Duncan test (p = 0.05). RESULTS: Specimens treated with dentin desensitizers (except Gluma) yielded significantly lower mean bond strengths than nontreated control specimens. The mean values in MPa (+/- SD) were: G1: 13.4 (6.2); G2: 13.2 (4.8); G3: 7.15 (4.3); G4: 7.21 (4.6). CONCLUSIONS: Among the desensitizing agents studied, only Gluma Desensitizer did not detrimentally influence the bond strength values. It is a useful material for dentin desensitization. PMID- 16708720 TI - Fracture resistance of fragmented incisal edges restored with fiber-reinforced composite. AB - PURPOSE: The aim of this study was to determine the static load-bearing capacity of fractured incisors restored with the conventional adhesive-composite technique or by using fiber-reinforced composites (FRC). MATERIALS AND METHODS: Twelve extracted sound maxillary incisors per group were prepared by cutting the incisal (one-third) part of the crown horizontally. Restorations were made using three techniques. Group A (control group) was restored by reattaching the original incisal edge to the tooth. Group B was restored using particulate filler composite (PFC). Group C was restored with PFC and FRC by adding a thin layer of FRC on the palatal surface of tooth. The bonding system used was the conventional etch system with primer and adhesive. All restored teeth were stored in water at room temperature for 24 h before they were statically loaded until fracture in a universal testing machine. Data were analyzed using ANOVA (p = 0.05). Failure modes were visually examined. RESULTS: Group A (reattaching fractured incisal edge) revealed the lowest load-bearing values, whereas preparation of the new incisal part with PFC revealed 148% higher load-bearing values compared to Group A. Group C (teeth restored with FRC) revealed a 254% higher load-bearing capacity than the control group. ANOVA revealed that the restoration technique significantly affected load-bearing capacity (p < 0.001). The failure mode in Groups A and B was debonding of the restoration from the adhesive interface, while in group C, 50% of the teeth fractured below the cementoenamel junction. CONCLUSION: These results suggest that an incisally fractured tooth restored with a combination of PFC and FRC provide the highest load-bearing capacity. PMID- 16708721 TI - Fatigue resistance of the bond of a glass-infiltrated alumina ceramic to human dentin. AB - PURPOSE: To evaluate the fatigue resistance of the bond between dentin and glass infiltrated alumina ceramic, using different luting protocols. MATERIALS AND METHODS: The null hypothesis is that the fatigue resistance varies with the luting strategy. Forty blocks of In-Ceram Alumina were prepared, and one surface of each block was abraded with 110-microm aluminum oxide particles. Then, the blocks were luted to flat dentin surfaces of 40 human third molars, using 4 different luting strategies (luting system [LS]/ceramic surface conditioning [CSC]) (n=10): (G1) [LS] RelyX-Unicem/[CSC] airborne abrasion with 110-microm Al2O3 particles; (G2) [LS] One-Step + Duo-Link (bis-GMA-based resin)/[CSC] etching with 4% hydrofluoric acid + silane agent; (G3) [LS] ED-Primer + Panavia F (MDP-based resin)/[CSC] Al2O3; (G4) [LS] Scotchbond1+RelyX-ARC (bis-GMA-based resin)/[CSC] chairside tribochemical silica coating (air abrasion with 30-microm SiOx particles + silane). After 24 h of water storage at 37 degrees C, the specimens were subjected to 106 fatigue cycles in shear with a sinusoidal load (0 to 21 N, 8 Hz frequency, 37 degrees C water). A fatigue survivor score was given, considering the number of the fatigue cycles until fracture. The failure modes of failed specimens were observed in a SEM. RESULTS: G3 (score = 5.9, 1 failure) and G4 (score = 6, no failures) were statistically similar (p = 0.33) and had significantly higher fatigue resistance than G1 (score = 3.9, 5 failures) and G2 (score = 3.7, 6 failures) (p < 0.03). SEM analysis of fractured specimens of G1 and G2 showed that almost all the failures were between ceramic and cement. CONCLUSION: The MDP-based resin cement + sandblasting with Al2O3 particles (G3) and bis-GMA-based resin cement + tribochemical silica coating (G4), both using the respective dentin bonding systems, were the best luting protocols for the alumina ceramic. The null hypothesis was confirmed. PMID- 16708722 TI - Microtensile bond strength between a quartz fiber post and a resin cement: effect of post surface conditioning. AB - PURPOSE: To test the bond strength between a quartz-fiber-reinforced composite post (FRC) and a resin cement. The null hypothesis was that the bond strength can be increased by using a chairside tribochemical silica-coating system. MATERIALS AND METHODS: Thirty quartz-FRCs (Light-Post) were divided into 3 groups according to the post surface treatment: G1) Conditioning with 32% phosphoric acid (1 min), applying a silane coupling agent; G2) etching with 10% hydrofluoric acid (1 min), silane application; G3) chairside tribochemical silica coating method (CoJet System): air abrasion with 30-microm SiOx-modified Al2O3 particles, silane application. Thereafter, the posts were cemented into a cylinder (5 mm diameter, 15 mm height) with a resin cement (Duo-Link). After cementation, the specimens were stored in distilled water (37 degrees C/24 h) and sectioned along the x and y axes with a diamond wheel under cooling (Lab-cut 1010) to create nontrimmed bar specimens. Each specimen was attached with cyanoacrylate to an apparatus adapted for the microtensile test. Microtensile testing was conducted on a universal testing machine (1 mm/min). The data obtained were submitted to the one-way ANOVA and Tukey test (alpha = 0.05). RESULTS: A significant influence of the conditioning methods was observed (p < 0.0001). The bond strength of G3 (15.14 +/ 3.3) was significantly higher than the bond strengths of G1 (6.9 +/- 2.3) and G2 (12.60 +/- 2.8) (p = 0.000106 and p = 0.002631, respectively). Notwithstanding the groups, all the tested specimens showed adhesive failure between the resin cement and FRC. CONCLUSION: The chairside tribochemical system yielded the highest bond strength between resin cement and quartz-fiber post. The null hypothesis was accepted (p < 0.0001). PMID- 16708723 TI - Adhesion between prefabricated fiber-reinforced posts and different composite resin cores: a microtensile bond strength evaluation. AB - PURPOSE: The aim of this study was to evaluate the bond strengths between various resin composites used as core materials (Multicore Flow, Ivoclar-Vivadent; Tetric Flow, Ivoclar-Vivadent; Filtek Flow, 3M-ESPE; Tetric Ceram, Ivoclar-Vivadent; Filtek Z250, 3M-ESPE), and an FRC post (FRC Postec Plus, Ivoclar-Vivadent) by means of the microtensile nontrimming technique. MATERIALS AND METHODS: Five experimental groups were used. For the microtensile nontrimming technique, 45 to 50 beam-shaped specimens per group were obtained from cylinders of core material, which had been built up around the post by progressively adding small increments of composite resin. Each specimen was loaded in tension until failure at either one of the two post/core interfaces present in each stick. The differences in interfacial bond strength amongthe groups were tested for statistical significance with the one-way ANOVA test, followed by the Dunnett test for post hoc comparisons. RESULTS: The measured bond strengths in MPa were 17.29 +/- 6.02 for FRC+MultiCore Flow, 16.37 +/- 6.92 for FRC+Tetric Flow, 13.14 +/- 5.35 for FRC + Filtek Flow, 12.38 +/- 4.34 for FRC + Tetric Ceram, and 10.75 +/- 5.43 for FRC + Filtek Z250. The statistical analysis revealed that MultiCore Flow achieved significantly higher bond strengths than Filtek Flow (p = 0.03), Tetric Ceram (p < 0.001), and Filtek Z250 (p < 0.001). The bond strength of Tetric Flow was significantly higher than that of Filtek Z250 (p = 0.003). CONCLUSION: For core buildup on a fiber post, dual-cure composites appear to be preferable to light curing composites. PMID- 16708724 TI - Two-year clinical evaluation of ormocer and nanofill composite with and without a flowable liner. AB - PURPOSE: The aim of this clinical follow-up study was to determine the clinical performance of ormocer and a new nanofill composite material, lined or not lined with flowable composites, after 2 years. MATERIALS AND METHODS: Occlusal restorations (n = 108) were placed in 54 patients with two symmetric restorations per patient. In one of the two restorations in each patient, cavities were first lined with the flowable composite material Admira Flow or Filtek Flow after applying the two-step total-etch one-bottle adhesive (Admira Bond) or single bond adhesive (Single Bond). Admira or Filtek Supreme was then applied. Restorations were examined using the USPHS modified Ryge criteria for retention, color matching, cavosurface marginal discoloration, anatomic form, marginal adaptation, surface texture, and secondary caries by two previously calibrated dentists. The overall performance of the restorations was evaluated by Wilcoxon signed ranks test. The chi-square test was used to determine differences in the performance of the restorations in which the flowable composite was applied first. RESULTS: There was no secondary caries or postoperative sensitivity in any of the restorations at the 6-month, 1-year, and 2-year follow-up examinations. After 2 years, only one Admira restoration had failed; all other restorations were available for evaluation. There were no statistically significant differences among the materials regardless of the prior use of flowable material (p > 0.05). There was no statistically significant difference between the restorations with and without the prior application of flowable materials (p > 0.05). CONCLUSION: The clinical performance of occlusal restorations using ormocer or nanofill composite did not benefit from the additional use of the flowable composite. Both of the restorative materials exhibited ideal clinical performance. PMID- 16708727 TI - Depressive illness--the need for a paradigm shift in its understanding and management. PMID- 16708725 TI - In vivo and in vitro evaluation of Class II composite resin restorations with different matrix systems. AB - PURPOSE: To investigate in vivo and in vitro Class II composite restorations performed with two matrix and wedge systems. MATERIALS AND METHODS: One hundred nine Class II restorations were performed in 23 patients, 59 with metallic matrices and wooden wedges (group 1) and 50 with polyester matrices and reflective wedges (group 2). All cavities were restored using Single Bond and P 60 (3M ESPE). In the metal matrix group, polymerization was performed from the occlusal, and in the polyester group through the reflective wedge. To assess microleakage, 40 proximal standard slot cavities were prepared in 20 noncarious human third molars. In the mesial cavity, the gingival margin was located at the enamel level, and in the distal cavity at the cementum/dentin. Specimens were randomly divided into two groups (n = 20) and restored with Single Bond, Z-250 (3M ESPE), and the same techniques used in the in vivo study: metal matrix/wooden wedge (group 1) and polyester matrix/reflective wedge (group 2). Specimens were thermocycled (500 times, 5 degrees C to 55 degrees C), then isolated with nail varnish and immersed in fuchsin for 8 h. Specimens were sectioned longitudinally and microleakage was assessed under magnification (40X) using a standard scoring system. RESULTS: Data were subjected to the chi-square test, Mann-Whitney U-test, and Kruskal-Wallis H-test. Matrix systems presented similar results in the clinical evaluation and the in vitro microleakage test of Class II composite restorations. Dye leakage was minimal at enamel margins, and statistically lower (p < 0.05) than at cementum/dentin margins. CONCLUSION: The different matrix systems had no influence on clinical performance or in vitro sealing ability of Class II composite restorations. PMID- 16708726 TI - Dengue fever in Malaysia: time for review? PMID- 16708728 TI - Tetanus in adults--a continuing problem: an analysis of 217 patients over 3 years from Delhi, India, with special emphasis on predictors of mortality. AB - Tetanus is still widely prevalent in many parts of the world especially in the developing countries. This study was conducted to assess the demographic and clinical features, treatment and outcome of tetanus in adults with a special emphasis on identification of predictors of mortality. This was a retrospective study of all adult patients admitted with tetanus over a three-year period from 1998 to 2000 at Lok Nayak Hospital, New Delhi, India. The demography, clinical features, treatment and complications of the patients were assessed and were evaluated against two possible outcomes- survival or death, to identify the predictors of mortality. There were 217 adults (148 males, 69 females) admitted with generalized tetanus during the study period. The mean age of the patients was 36.08 +/- 18.84 years. In most instances (63.5%), wounds / injuries served as the source of infection. Overall 31.8% had mild, 21.7% had moderate and 46.5% had severe disease at presentation. Respiratory failure and dysautonomia were the frequently observed complications. Of the 217 patients, 82 died - a mortality rate of 37.78%. Parameters that were significantly associated with increased mortality were: age (especially > 60 years); shorter duration of symptoms - trismus, rigidity and dysphagia; severe disease at presentation and shorter period of onset. Tetanus is a preventable disease associated with a high fatality rate. Universal coverage of immunization programme and appropriate wound prophylaxis is the corner stone of tetanus eradication. The treatment of tetanus is difficult and requires specialized and intensive care. The presence of parameters indicating an adverse outcome at the beginning in the patients will help identify the subset of patients that need more intensive management especially in resource poor nations. PMID- 16708729 TI - The prevalence of depression among elderly warded in a tertiary care centre in Wilayah Persekutuan. AB - Depression is characterized by symptoms like disturbance in behavior, cognition and mood. This is commonly known to affect people aged 60 years and above especially those who are also afflicted with illnesses. The objectives of this study were to determine the prevalence of depression and its associated factors among the elderly in a tertiary care centre in Wilayah Persekutuan. A cross sectional study design was done. Stratified cluster sampling method was used to select the respondents. All elderly patients were selected from the orthopaedic, surgical, gynaecology and medical wards in a government hospital in Wilayah Persekutuan. A 30-item Geriatric Depression Scale questionnaire was used as a screening instrument. Out of the 246 elderly subjects, 198 were interviewed giving a response rate of 80.5%. The results showed that 54% of the elderly respondents were found to have depressive symptoms. Age (p=0.022), sex (p=0.008), ethnicity (p=0.022) and functional disabilities in bathing (p=0.001), grooming (p=0.007), dressing (p= 0.007), using the toilet (p=0.002), transferring from bed to chair and back (p=0.000), mobility (p=0.000) and climbing stairs (p=0.000) were all found to be significantly associated with depression among the elderly respondents. The outcome of this study will have an important impact on the implementation of the health policy for the elderly patients admitted to hospitals. PMID- 16708730 TI - Repeated tourniquet testing as a diagnostic tool in dengue infection. AB - Prospective evaluation of repeated standard tourniquet testing as a diagnostic indicator of dengue infection was done. Included were patients admitted to a children's hospital in Kuala Lumpur on a clinical suspicion of dengue infection based on the World Health Organization (WHO) criteria. A standard method of tourniquet was performed on 79 patients on a daily basis following admission. subjects and negative in the remaining 14 subjects. Fifty-eight subjects were serologically confirmed cases, 4 indeterminate and the remaining 17 subjects had negative serology. For diagnostic classification, 13 had dengue fever, 49 with dengue haemmorhagic fever (DHF) while 17 had non-dengue infection. The sensitivity and specificity of the tourniquet test was 82.8% and 23.5% respectively. The positive predictive value (PPV) was 78.7% while the negative predictive value (NPV) was 28.6%. In addition, the tourniquet test aided in the diagnosis of one-fifth of patients with DHF, who presented with a positive tourniquet test as the only bleeding manifestation. It seems that in a hospital setting, the tourniquet test adds little to the diagnosis of dengue infection/DHF. A positive tourniquet test, repeatedly performed, was found clinically useful as a preliminary screening test in dengue infection as recommended by WHO. However, it was not very specific and had a high false positive rate. PMID- 16708731 TI - An open label comparative study of glimepiride versus repaglinide in type 2 diabetes mellitus Muslim subjects during the month of Ramadan. AB - This study was conducted to compare the treatment efficacy between a prandial glucose regulator, repaglinide and a new sulphonylurea, glimepiride in Muslim Type 2 diabetic patients who practice Ramadan fasting. Forty-one patients, previously treated with a sulphonylurea or metformin, were divided to receive either repaglinide (n=20, preprandially three-times daily) or glimepiride (n=21, preprandially once daily) 3 months before the month of Ramadan. During Ramadan, patients modified their eating pattern to two meals daily, and the triple doses of repaglinide were redistributed to two preprandial doses. Four point blood glucose monitoring were performed weekly during the month of Ramadan and the subsequent month. Measurements of the 4-point blood glucose were significantly lower in the glimepiride group compared to the repaglinide group both during and after Ramadan. The glycaemic excursion was better in the morning for the repaglinide group and better in the afternoon and evening for the glimepiride group during the Ramadan period. There was no statistically significant difference in the incidence of hypoglycaemia between the two groups during and after Ramadan. There was no difference in the glycaemic excursion post-Ramadan. The longer duration of action of glimepiride may offer an advantage over repaglinide during the 13.5 hours of fast in Ramadan for diabetic patients. PMID- 16708732 TI - Neuroimaging features of tuberculous meningitis. AB - Tuberculous meningitis leads to a high mortality rate. However, it responds well to chemotherapy if the treatment is started early. Neuroimaging is one of the most important initial investigations. There were 42 patients diagnosed with tuberculous meningitis in Kuala Lumpur Hospital based on clinical criteria, cerebrospinal fluid analysis and response to anti-tuberculous treatment over a 7 year period. Relevant information was obtained from patients' medical case notes and neuroimaging findings were evaluated. Male to female ratio was 3:1. The three major ethnics and the immigrant groups in Malaysia were represented in this study. The majority of the cases involved the Malays followed by immigrants, Chinese and Indians. The patients' age ranged from 18 to 62 years old with the mean age of 34.4 years. There were 95.2% (n = 40) of patients who presented with various neuroimaging abnormalities and only 2 (4.8%) patients had normal neuroimaging findings. Hydrocephalus and meningeal enhancement were the two commonest neuroimaging features. Other features include infarction, enhancing lesion, tuberculoma, abcess, oedema and calcification. Contrasted CT scan is an adequate neuroimaging tool to unmask abnormal findings in tuberculous meningitis. PMID- 16708733 TI - Prevalence of smoking and drinking habits among Form Six students in Petaling District, Selangor. AB - Smoking and alcohol intake patterns may persist from adolescence to adulthood. The aims of this study are to determine the prevalence and factors associated with smoking and drinking habits among Form Six students. This was a cross sectional study conducted in January 2003 among Form Six students from government schools in the Petaling District, Selangor. A hundred and thirty six self administered questionnaires were distributed to students selected through multistage stratified sampling. Response rate in this study was 90.1% (136/151). The respondents were mainly Chinese 60 (44.1%) and female 88 (64.7%). The prevalence of smoking was 22.8%, whereas the prevalence of alcohol intake was 47.8%. Only 33.1% of the respondents practiced undetermined activities and 1.5% have undetermined characters. There were significant associations between smoking and males (Adjusted OR 2.56, 95% CI 1.02-6.43) and smoking and alcohol intake (Adjusted OR 2.74, 95% CI 1.11-6.78). Alcohol intake has significant negative association with Malays (Adjusted OR 0.83, 95% CI 0.03-0.27). Smoking habits among adolescents were associated with males and alcohol intake. However, only alcohol intake was negatively associated with Malays. Program interventions to reduce behavioral problems, particularly smoking and alcohol intake should be emphasized. PMID- 16708734 TI - Estimates and distribution of body mass index in a sample of Malaysian adolescents. AB - The purpose of this study was to report on the estimates and distribution of body mass index in a sample of Malaysian adolescents. The study utilized a cross sectional design and multi-stage random sampling of secondary schools to select 5 urban and 9 rural schools in Kedah and Penang. A total of 6555 male and female adolescents (11-15 years old) of Malay, Chinese and Indian ethnic groups were measured for weights and heights for body mass index calculation. Information on household demographic and socioeconomic were obtained from parents through self administered questionnaires. Analyses of body mass index distribution by location, ethnicity, gender and age were conducted using Chi-square test of SPSS 11.5. More of the rural (12.1%) and urban (19.4%) adolescents were underweight and overweight, respectively. While in all ethnic, gender and age groups, rural adolescents were more likely to be underweight, more of the urban adolescents were overweight. The prevalence of underweight was highest among the Indians (19.2%) and lowest in Chinese (7.2%). The prevalence of overweight in the three ethnic groups was in the range of 18-19%. More male than female adolescents were underweight (15% vs 7.8%) and overweight (19.5% vs 16.7%). Consistent patterns were also observed across location, ethnic and age groups. As age increased, the prevalence of overweight decreased across the ethnic and gender groups. The reported findings can serve as current reference on body mass index distribution of Malaysian adolescents and a basis for future efforts in health and nutrition interventions for Malaysian children and adolescents. PMID- 16708735 TI - A prospective comparison of percutaneous endoscopic gastrostomy and nasogastric tube feeding in patients with acute dysphagic stroke. AB - Dysphagia following stroke is common problem and is of particular concern because of its potental for malnutrition. Nasogastric (NG) and percutaneous endoscopic gastrostomy (PEG) tube feeding are recognized methods for nutritional support for patients with persistent neurologic dysphagia. However, the former is associated with tube dislodgement and blockage that might compromise the patients' nutritional status. There have been few randomized prospective studies to date comparing the efficacy and safety of these 2 modes of dysphagia management in stroke patients. The objective of this study was to compare PEG with NG tube feeding after acute dysphagic stroke in terms of nutritional status and treatment failure. This was a randomized prospective clinical trial. A total of 23 consecutive patients who fulfilled the criteria were recruited from the medical wards in Hospital Universiti Kebangsaan Malaysia. The diagnosis of stroke (acute cerebral infarct) was based on clinical and brain computed tomographic (CT scan) findings; and the diagnosis of dysphagia was done clinically by using the 'swallowing test'. At recruitment, upper-arm skin fold thickness (triceps and biceps) and mid-arm circumference were measured; and blood was drawn for serum albumin level. They were then followed up at 4 weeks where the above tests were repeated. A total of 22 patients completed the study (12 patients in the NG group and 10 patients in the PEG group). Serum albumin levels (p = 0.045) were significantly higher in the PEG as compared to the NG group at 4 weeks post intervention. There were statistically significant improvements in serum albumin level (p = 0.024) in the PEG group; and statistically significant reductions in serum albumin level (p = 0.047) in the NG group 4 weeks after the intervention. However, there were no significant differences in anthropometric parameters between the two groups and no significant changes in these parameters for each group 4 weeks after the intervention. Treatment failure occurred in 5 out of 10 patients (50.0%) in the NG group, but none in PEG group (p = 0.036). PEG tube feeding is more effective than NG tube feeding in improving the nutritional status (in terms of the serum albumin level) of patients with dysphagic stroke. NG tube feeding, in fact, reduced the nutritional status (in terms of the serum albumin level) of the patients. PMID- 16708736 TI - Correlation between fasting plasma glucose, post prandial glucose and glycated haemoglobin and fructosamine. AB - This study was done to determine the correlation between glucose monitoring by fasting blood glucose or 2 hours postprandial blood glucose with HbA1c and fructosamine in type 2 diabetic patients. A total of 82 patients from the Primary Care Clinic were enrolled in the study. Fasting blood was drawn for fasting plasma glucose (FPG), glycated haemoglobin (HbA1c) and fructosamine. Two hours after a standard breakfast, blood was again drawn for prandial plasma glucose (PPG). Both PPG and FPG significantly correlated with both HbA1c and fructosamine but PPG showed better correlation to HbA1c than FPG (r= 0.604 vs.0.575) whereas that of FPG and PPG were equally correlated to fructosamine (r= 0.566 vs. 0.551). In predicting good glycaemic control (HbA1c < 7.0%), the sensitivity, specificity and positive predictive value of PPG were 75.0%, 80.6% and 82.5% whereas FPG were 81.8%, 58.3% and 70.6% respectively. These results show that PPG correlated better than FPG to HbA1c and both equally correlated to fructosamine levels. Thus, PPG predicted overall glycaemic control better than FPG. Compared to HbA1c, fructosamine correlated least well with mean glucose profiles. Hence, using HbAlc in monitoring overall glycaemic control is better than fructosamine. PMID- 16708737 TI - The translabyrinthine approach for acoustic neuroma and its common complications. AB - A retrospective analysis of 15 cases intracanalicular acoustic neuroma that undergone tumour excision by translabyrinthine approach spanning from August 1996 until December 2002 is presented. The main presenting complaints are unilateral hearing loss (100%) and tinnitus (86.7%). The mean age of presentation was 48.5 years old. Magnetic resonance imaging is the most important investigation tool to diagnose acoustic neuroma. At six months post operatively, the facial nerve was normal or near normal (grade I and II) in 46.6%, grade III to IV in 46.6% and grade V to VI in 6.7% of the cases respectively. There were also four cases of post operative cerebrospinal fluid leak, which was successfully managed with conservative measures. The translabyrinthine approach is the most familiar surgical technique employed by otologist. It is the most direct route to the cerebellopontine angle and internal auditory canal. It requires minimum cerebellar retraction. However, it sacrifices any residual hearing in the operated ear. PMID- 16708738 TI - Postnatal depression and socio-cultural practices among postnatal mothers in Kota Bahru, Kelantan, Malaysia. AB - This is a cross sectional study to determine the relationship of postnatal depression (PND) and socio-cultural practices post-delivery among women in Kota Bharu, Kelantan. Four hundred and twenty one pregnant women were screened for depression between 36 - 42 weeks of pregnancy, 1 week and 4 - 6 weeks postpartum using Edinburgh Postnatal Depression Scale (EPDS). The women also completed questionnaires on socio-demography, psychosocial support and traditional postnatal care. The prevalence of PND at 4-6 weeks postpartum was 20.7%. Depressive symptoms at the end of pregnancy (p<0.05) and one week postpartum (p<0.05), worry about the baby (p<0.05), use of traditional medication (p<0.05) and traditional massage (p<0.05) were significantly associated with PND. PMID- 16708739 TI - Massive epistaxis secondary to pseudoaneurysm of internal carotid artery. AB - Post-traumatic pseudoaneurysms of internal carotid arteries are uncommon. The patients may present with massive epistaxis due to rupture of the aneurysm into the sphenoid sinus. Early diagnosis and treatment is mandatory as the likelihood of exsanguinations increases with each subsequent episode of epistaxis. The clinical features of unilateral blindness and massive epistaxis after head injury should indicate the diagnosis. The high mortality of this entity underlines the importance of early angiography in these patients to confirm this diagnosis. We present 3 cases of post-traumatic aneurysm of the ICA. PMID- 16708740 TI - Mucinous adenocarcinoma arising from chronic perianal fistula. AB - Perianal mucinous adenocarcinoma is a rare tumor which may be associated with long-standing chronic perianal sepsis. Early diagnosis is challenging and is based on a high index of clinical suspicion and specific histological features. Definitive treatment is surgical, in the form of an abdomino-perineal resection. We hereby describe a case of a perianal mucinous adenocarcinoma arising from long standing recurrent perianal fistula and complement this with a brief review of the literature pertaining in particular to the management of this condition. PMID- 16708741 TI - Haemophagocytic syndrome presenting as pyrexia of unknown origin. AB - Haemophagocytic syndrome (HPS) should be included in the differential diagnosis of pyrexia of unknown origin (PUO). The hallmark of HPS is the accumulation of activated macrophages that engulf haematopoietic cells in the reticuloendothelial system. We describe a patient with unexplained fever in which a final diagnosis of HPS was established in a bone marrow study. PMID- 16708742 TI - Acquired nasal posterior choanal atresia: postradiotherapy. AB - Reported cases of acquired posterior choanal atresia are very few in the English literature. A case of acquired posterior choanal atresia post radiotherapy is reported which was treated by endonasal endoscopic repair using microdebrider with untoward effect. PMID- 16708743 TI - A case of Hodgkin's disease presenting with recurrent laryngeal nerve palsy and tracheoesophageal fistula. AB - Malignancy is the most common cause of tracheoesophageal fistulas. The malignancies commonly implicated in the development of tracheoesophageal fistulas are primary bronchial or esophageal carcinomas. Hodgkins disease rarely leads to such a fistula. We report a case of Hodgkin's disease with a tracheoesophageal fistula as well as a left recurrent nerve palsy at presentation. This presentation has no precedent in literature. The patient was treated with systemic chemotherapy and involved field radiotherapy. On follow up 1 year after the completion of treatment, he was clinically and radiologically disease free. PMID- 16708744 TI - Neonatal posterior fossa haemorrhage associated with vacuum extractor. AB - We report one case of posterior fossa intracranial haemorrhage in a full-term Malay baby boy following vacuum assisted delivery. The patient, a term baby boy was delivered by a vacuum extraction and later developed signs of increased intracranial pressure 72 hours after birth. Computed tomography (CT) of the brain showed a posterior fossa intracranial haemorrhage with acute obstructive hydrocephalus. He was initially treated with isolated ventricular shunting which later caused an upward cerebellar herniation. An immediate suboccipital craniectomy for evacuation of cerebellar haematoma was performed which resulted in a gradual recovery. PMID- 16708745 TI - Rare bone disorder affecting the temporal bone. AB - Fibrous dysplasia is an uncommon benign disorder of unknown etiology. Rarely, it presents isolated in the temporal bone. We present three cases of monostotic fibrous dysplasia that involved the entire temporal bone. PMID- 16708746 TI - Rhino-orbito-cerebral mucormycosis: a treatment dilemma. AB - A case of rhino-orbito-cerebral mucormycosis is presented showing its aggressive nature and progression of disease. The typical clinical features, neuroimaging and histological findings are highlighted in this report. Amphotericin B and surgical debridement remain the mainstay of treatment. However, associated co morbidities need to be addressed. PMID- 16708747 TI - High-flux haemodialysis treatment as treatment for carbamazepine intoxication. AB - Acute severe intoxication with carbamazepine is associated with seizures, coma and respiratory depression. Traditionally, charcoal haemoperfusion is used to remove the drug. We present a case of carbamazepine intoxication, successfully treated with three hours of high-flux haemodialysis. Thus, haemodialysis using high-flux membranes is a feasible and effective therapeutic option for carbamazepine intoxication. PMID- 16708748 TI - Intranasal use of QuickClot in a patient with uncontrollable epistaxis. AB - A 60-year-old man who presented with nasopharyngitis developed uncontrollable epistaxis following a punch biopsy of the nasopharynx. QuickClot was successfully used to arrest the haemorrhage under general anaesthesia after the usual methods employed to secure haemostasis failed. The haemostatic plug was successfully removed a week later after control of the infection. This case represents the first reported intranasal use of QuickClot. We describe our experience and a literature review on this haemostatic agent. PMID- 16708750 TI - Clearing the air about "the haze". PMID- 16708749 TI - Technique of anaesthesia in pulmonary hypertension and thrombophilia in early pregnancy. AB - Pulmonary hypertension in pregnancy is a rare condition but is associated with a high mortality. We report the case of a 29 year old female in early pregnancy with Protein C and S deficiency with recurrent deep venous thrombosis and pulmonary embolism and subsequent secondary pulmonary hypertension. The patient was counselled and consented for termination of pregnancy with tubal sterilization. She was administered continuous spinal anaesthesia with invasive monitoring. The successful anaesthetic management of this condition is described. PMID- 16708751 TI - Factors influencing post-operative hyperglycaemia in type 2 diabetes mellitus patients managed without intra-operative insulin. AB - Surgery induces a 'stress' state leading to post-operative hyperglycaemia. To investigate this effect on patients with Type 2 Diabetes Mellitus, we reviewed the records of 50 diabetic patients who underwent surgery without intraoperative insulin. Demographic features together with pre-operative and post-operative blood glucose readings were noted. 27.3% of patients with well controlled pre operative blood glucose levels developed post-operative hyperglycaemia. In contrast, 84.6% of patients with poorly controlled levels developed the same. Poor control of blood glucose and duration of operation were the only significant predictors of post-operative hyperglycaemia. PMID- 16708752 TI - Use of complementary medicine amongst asthmatic patients in primary care. AB - Complementary Medicine (CM) usage amongst asthmatic patients was studied. Eighty eight patients, selected by systematic random sampling in two public polyclinics in April/May 2004, were interviewed. They completed a structured pre-tested questionnaire. Forty-one percent were using CM, majority (64%) together with conventional therapy. Eighty-one percent did not inform their physicians of their CM usage. More Malays were using CM which included nutritional supplements, herbs, yoga, homoeopathy, reflexology and massage. PMID- 16708753 TI - Community-acquired pneumonia in Malaysian patients: addition of macrolide and the use of BTS "curb" index to assess severity. PMID- 16708754 TI - Ovulation induction and ovarian cancer: is there a link? PMID- 16708755 TI - Following your passions. PMID- 16708756 TI - Utility of cell-permeable peptides for enhancement of virus-mediated gene transfer to human tumor cells. PMID- 16708757 TI - Software to perform automated comparisons of pair-wise percent identities for microbial species. PMID- 16708758 TI - Differential recovery of biotinylated microbial proteins using monomeric or polymeric avidin. PMID- 16708759 TI - Affinity-capture protease reactor for single-step production and purification of antiangiogenic plasminogen fragment from human plasma. PMID- 16708761 TI - Application of in situ ductal perfusion to facilitate isolation of high-quality RNA from mouse pancreas. AB - A technique to isolate high-quality intact RNA from murine pancreas is described. This technique involves in situ ductal perfusion of the pancreas with an RNase inhibitor prior to removal of the organ for RNA extraction. In this way, the pancreatic RNases are inhibited in situ allowing good yields of intact RNA, suitable for studies on pancreatic gene transcription by real-time PCR or microarray analysis, to be obtained in a reliable way. PMID- 16708760 TI - Functional reconstitution of Beta2-adrenergic receptors utilizing self-assembling Nanodisc technology. AB - Integral membrane G protein-coupled receptors (GPCRs) compose the single most prolific class of drug targets, yet significant functional and structural questions remain unanswered for this superfamily. A primary reason for this gap in understanding arises from the difficulty of forming soluble, monodisperse receptor membrane preparations that maintain the transmembrane signaling activity of the receptor and provide robust biophysical and biochemical assay systems. Here we report a technique for self-assembling functional beta2-adrenergic receptor (beta2AR) into a nanoscale phospholipid bilayer system (Nanodisc) that is highly soluble in aqueous solution. The approximately 10-nm nanobilayer particles contain beta2AR in a native-like phospholipid bilayer domain of approximately 100 phospholipid molecules circumferentially bound by a membrane scaffold protein (MSP). The resulting construct allows for access to the physiologically intracellular and extracellular faces of the receptor and thus allows unrestricted access of antagonists, agonists, and G proteins. These Nanodisc-solubilized GPCRs can be directly purified by normal chromatographic procedures. We define the resultant Nanodisc-embedded monomeric beta2AR by antagonist and agonist binding isotherms and demonstrate faithful G protein coupling. PMID- 16708762 TI - Yeast-based functional genomics and proteomics technologies: the first 15 years and beyond. AB - Yeast-based functional genomics and proteomics technologies developed over the past decade have contributed greatly to our understanding of bacterial, yeast, fly, worm, and human gene functions. In this review, we highlight some of these yeast-based functional genomic and proteomic technologies that are advancing the utility of yeast as a model organism in molecular biology and speculate on their future uses. Such technologies include use of the yeast deletion strain collection, large-scale determination of protein localization in vivo, synthetic genetic array analysis, variations of the yeast two-hybrid system, protein microarrays, and tandem affinity purification (TAP)-tagging approaches. The integration of these advances with established technologies is invaluable in the drive toward a comprehensive understanding of protein structure and function in the cellular milieu. PMID- 16708763 TI - Reverse transcription using random pentadecamer primers increases yield and quality of resulting cDNA. AB - Reverse transcription of RNA is an invaluable method for gene expression analysis by real-time PCR or microarray methods. Random primers of varying lengths were compared with respect to their efficiency of priming reverse transcription reactions. The results showed that 15-nucleotide-long random oligonucleotides (pentadecamers) consistently yielded at least 2-fold as much cDNA as did random hexamers using either poly(A) RNA or an amplified version of messenger RNA (aRNA) as a template. The cDNA generated using pentadecamers did not differ in size distribution or the amount of incorporated label compared with cDNA generated with random hexamers. The increased efficiency of priming using random pentadecamers resulted in reverse transcription of >80% of the template aRNA, while random hexamers induced reverse transcription of only 40% of the template aRNA. This suggests a better coverage of the transcriptome when using random pentadecamers over random hexamers. Using the same amount of aRNA as starting material, random pentadecamer-primed reactions resulted in 11-fold more genes being detected in whole transcriptome DNA microarray experiments than random hexamer-primed reactions. The results indicate that random pentadecamers can replace random hexamers in reverse transcription reactions on both poly(A) RNA and amplified RNA, resulting in higher cDNA yields and quality. PMID- 16708764 TI - Multiplex dosage pyrophosphorolysis-activated polymerization: application to the detection of heterozygous deletions. AB - Large heterozygous chromosomal deletions and gene duplications are important classes of mutations that are generally missed by standard PCR amplification and sequencing. Multiplex dosage pyrophosphorolysis-activated polymerization (MD PAP), a derivative of PAP, was utilized to detect these types of mutations. PAP is a method for nucleic acid amplification in which 3' blocked oligonucleotides (P*) are activated by pyrophosphorolysis when annealed to the target template and subsequently extended. A key advantage to this technology is that PAP reactions produce little or no primer-dimer or false priming. As a result of this enhanced specificity, MD-PAP is easy to optimize. Herein, we utilize MD-PAP to determine gene dosage of each exon of the human factor IX gene by comparison with one endogenous internal control from the ATM gene. Estimated dosage is proportional to the actual template copy number over a minimum dynamic range from 1 to 16 copies. A blinded analysis detected 100% of 43 heterozygous deletions of exons in the human factor IX gene. PMID- 16708765 TI - The evolution of research: meeting the needs of systems medicine, part 1. PMID- 16708766 TI - Reduction of tinnitus by spinal manipulation in a patient with presumptive rotational vertebral artery occlusion syndrome: a case report. PMID- 16708767 TI - Essential fatty acids. PMID- 16708768 TI - The add-on effects of Gynostemma pentaphyllum on nonalcoholic fatty liver disease. AB - CONTEXT: Other than weight reduction by dieting or physical activity, there are no well-documented medical treatments for fatty liver disease. OBJECTIVE: To evaluate the efficacy of the add-on Gynostemma pentaphyllum (GP) in research subjects with nonalcoholic fatty liver disease. DESIGN: A randomized, single blind, controlled clinical trial. SETTING: Hospital-based clinic. PATIENTS: Fifty six research subjects who were diagnosed with nonalcoholic fatty liver by abdominal ultrasound scanning. INTERVENTIONS: The treatment group and the control group followed a controlled diet for 2 months. After 2 months, the treatment group continued to diet and received 80 mL GP extraction for 4 months; the control group continued to diet and received a placebo capsule for 4 months. MAIN OUTCOME MEASURES: Body mass index (BMI), biochemistry data, and fatty liver score were measured at baseline, at 2 months, and at 6 months. RESULTS: After 2 months of dieting, BMI and most biochemistry data decreased in both study groups. There were no significant differences in BMI or biochemistry data at month 2 between the 2 study groups. At month 6, BMI, triglyceride, aspartate aminotransferase (AST), alanine aminotransferase, alkaline phosphatase, insulin (ALP), insulin resistance index (HOMA-IR), and fatty liver score were reduced in both groups. The treatment group saw significant reductions in BMI, AST, ALP, insulin, and HOMA-IR, however. Changes in uric acid levels in the 2 groups from month 2 to month 6 were statistically significant (P = .028) CONCLUSION: GP is an effective adjunct treatment to diet therapy for patients with nonalcoholic fatty liver disease. PMID- 16708769 TI - Mercury elimination with oral DMPS, DMSA, vitamin C, and glutathione: an observational clinical review. AB - Tissue mercury levels in humans have increased during the past 50 years to an alarming concentration, with possible deleterious effects that may involve neurological, cardiovascular, and immunological pathology. This article reviews the protocol for the use of oral 2,3-dimercaptopropane-1-sulfonate (DMPS) and oral meso-2, 3-dimercaptosuccinic acid (DMSA) in combination with intravenous glutathione and high-dose vitamin C for treatment of high-level mercury. This protocol yielded an average 69% reduction of urine mercury by provocation analysis. PMID- 16708770 TI - Are health services research methods a match for CAM? AB - BACKGROUND: Many complementary and alternative medicine (CAM) researchers believe that a new set of research methods is needed to study CAM. Health services research (HSR) is listed as a new research goal in the National Center for Complementary and Alternative Medicine 2005-2009 strategic plan. Does HSR offer solutions to the research needs of CAM? This article presents an overview of HSR, a summary of the HSR studies that have been published in CAM, and discussions of what HSR might offer CAM and what studies of CAM might offer conventional HSR. DISCUSSION: Our literature search, which was performed at the University of Arizona, found 84 published CAM HSR studies. A comparison of the search results mostly surveys of CAM users or providers and economic outcome studies-with the breadth of topics that may be studied using HSR, showed that research in this field has not reached its potential. More work, including studies of interactions between patient, provider, and the system as well as patient-centered outcomes studies, is needed. Several areas in which CAM could benefit from HSR were identified and discussed. These included studies supporting the integration of CAM and conventional medicine, insurance coverage for CAM therapies, the development practice guidelines, and studies of the effectiveness and cost effectiveness of CAM therapies and whole medical systems. The study of CAM, especially studies focusing on patients' attraction to CAM, may also help expand the topics and methods of conventional HSR. SUMMARY: HSR methods may address many of CAM's methodological challenges and improve clinical, patient-centered, and economic outcomes across all systems of care-both conventional and CAM. PMID- 16708771 TI - Rachel Naomi Remen, MD: recovering the soul of medicine. Interview by Karolyn A. Gazella and Suzanne Snyder. PMID- 16708772 TI - An early "Atkins' Diet": RA Fisher analyses a medical "experiment". AB - A study on vitamin absorption which RA Fisher analysed for WRG Atkins and co authored with him is critically examined. The historical background as well as correspondence between Atkins and Fisher is presented. PMID- 16708773 TI - On Bayesian inference for the K function. AB - The K function is a summary of spatial dependence in spatial point processes. In practice one observes a realization of the spatial point process, called a spatial point pattern. Although the K function of a spatial point process is typically unknown, several estimators of the process K function have been put forth. These estimators, however, are based upon empirical averages; the complicated distributional properties of the estimators unfortunately complicates interval estimation. In this paper, we propose a Bayesian inferential framework, allowing inference for the K function of the spatial point process (including interval estimation). Of particular interest is the unique use of the posterior predictive distribution to (efficiently) enable such inferences. To demonstrate our technique, the well known Swedish pine sapling data (Strand, 1972) is analyzed, including a discussion on evaluating model fit. PMID- 16708774 TI - Bayesian analysis for the meiosis I non-disjunction fraction in numerical chromosomal anomalies. AB - The main causes of numerical chromosomal anomalies, including trisomies, arise from an error in the chromosomal segregation during the meiotic process, named a non-disjunction. One of the most used techniques to analyze chromosomal anomalies nowadays is the polymerase chain reaction (PCR), which counts the number of peaks or alleles in a polymorphic microsatellite locus. It was shown in previous works that the number of peaks has a multinomial distribution whose probabilities depend on the non-disjunction fraction F. In this work, we propose a Bayesian approach for estimating the meiosis I non-disjunction fraction F. in the absence of the parental information. Since samples of trisomic patients are, in general, small, the Bayesian approach can be a good alternative for solving this problem. We consider the sampling/importance resampling technique and the Simpson rule to extract information from the posterior distribution of F. Bayes and maximum likelihood estimators are compared through a Monte Carlo simulation, focusing on the influence of different sample sizes and prior specifications in the estimates. We apply the proposed method to estimate F. for patients with trisomy of chromosome 21 providing a sensitivity analysis for the method. The results obtained show that Bayes estimators are better in almost all situations. PMID- 16708776 TI - A renewed approach to the nonparametric analysis of replicated microarray experiments. AB - DNA-microarrays find broad employment in biochemical research. This technology allows the monitoring of the expression levels of thousands of genes at the same time. Often, the goal of a microarray study is to find differentially expressed genes in two different types of tissue, for example normal and cancerous. Multiple hypothesis testing is a useful statistical tool for such studies. One approach using multiple hypothesis testing is nonparametric analysis for replicated microarray experiments. In this paper we present an improved version of this method. We also show how p-values are calculated for all significant genes detected with this testing procedure. All algorithms were implemented in an R-package, and instructions on it's use are included. The package can be downloaded at http://www.statistik.unidortmund.de/de/content/einrichtungen/lehrstuehle/personen jung.html PMID- 16708775 TI - Bayesian inference for a correlated 2 x 2 table with a structural zero. AB - We develop a new Bayesian approach to interval estimation for both the risk difference and the risk ratio for a 2 x 2 table with a structural zero using Markov chain Monte Carlo (MCMC) methods. We also derive a normal approximation for the risk difference and a gamma approximation for the risk ratio. We then compare the coverage and interval width of our new intervals to the score-based intervals over various parameter and sample-size configurations. Finally, we consider a Bayesian method for sample-size determination. PMID- 16708777 TI - Regression as a method to predict copy numbers in comparative genomic hybridization studies on bacteria. AB - Comparative genomic hybridizations (CGH) using microarrays are performed with bacteria in order to determine the level of genomic similarity between various strains. The microarrays applied in CGH experiments are constructed on the basis of the genome sequence of one strain, which is used as a control, or reference, in each experiment. A strain being compared with the known strain is called the unknown strain. The ratios of fluorescent intensities obtained from the spots on the microarrays can be used to determine which genes are divergent in the unknown strain, as well as to predict the copy number of actual genes in the unknown strain. In this paper, we focus on the prediction of gene copy number based on data from CGH experiments. We assumed a linear connection between the log2 of the copy number and the observed log2-ratios, then predictors based on the factor analysis model and the linear random model were proposed in an attempt to identify the copy numbers. These predictors were compared to using the ratio of the intensities directly. Simulations indicated that the proposed predictors improved the prediction of the copy number in most situations. The predictors were applied on CGH data obtained from experiments with Enterococcus faecalis strains in order to determine copy number of relevant genes in five different strains. PMID- 16708778 TI - Assessing the amount of heterogeneity in random-effects meta-analysis. AB - In a random-effects meta-analysis, a new confidence interval for the heterogeneity parameter is proposed. With this interval, the amount of heterogeneity in a meta-analysis can be assessed so that it can be judged whether the pooling of the estimates is meaningful. Through suitable corrections of the lower bound, based on the treatment effect measure of interest, the resulting interval yields satisfactory results with respect to the predefined confidence coefficient. Lower and upper bound of the interval can be used for one-sided hypothesis testing on the amount of the underlying between-trial variability. PMID- 16708779 TI - Estimating correlation coefficient between two variables with repeated observations using mixed effects model. AB - We estimate the correlation coefficient between two variables with repeated observations on each variable, using linear mixed effects (LME) model. The solution to this problem has been studied by many authors. Bland and Altman (1995) considered the problem in many ad hoc methods. Lam, Webb and O'Donnell (1999) solved the problem by considering different correlation structures on the repeated measures. They assumed that the repeated measures are linked over time but their method needs specialized software. However, they never addressed the question of how to choose the correlation structure on the repeated measures for a particular data set. Hamlett et al. (2003) generalized this model and used Proc Mixed of SAS to solve the problem. Unfortunately, their method also cannot implement the correlation structure on the repeated measures that is present in the data. We also assume that the repeated measures are linked over time and generalize all the previous models, and can account for the correlation structure on the repeated measures that is present in the data. We study how the correlation coefficient between the variables gets affected by incorrect assumption of the correlation structure on the repeated measures itself by using Proc Mixed of SAS, and describe how to select the correlation structure on the repeated measures. We also extend the model by including random intercept and random slope over time for each subject. Our model will also be useful when some of the repeated measures are missing at random. PMID- 16708780 TI - A simple imputation method for longitudinal studies with non-ignorable non responses. AB - Missing data are a common problem in longitudinal studies in the health sciences. Motivated by data from the Muscatine Coronary Risk Factor (MCRF) study, a longitudinal study of obesity, we propose a simple imputation method for handling non-ignorable non-responses (i.e., when non-response is related to the specific values that should have been obtained) in longitudinal studies with either discrete or continuous outcomes. In the proposed approach, two regression models are specified; one for the marginal mean of the response, the other for the conditional mean of the response given non-response patterns. Statistical inference for the model parameters is based on the generalized estimating equations (GEE) approach. An appealing feature of the proposed method is that it can be readily implemented using existing, widely-available statistical software. The method is illustrated using longitudinal data on obesity from the MCRF study. PMID- 16708781 TI - Analysis of longitudinal multinomial outcome data. AB - Analysis of categorical outcomes in a longitudinal study has been an important statistical issue. Continuous outcome in a similar study design is commonly handled by the mixed effects model. The longitudinal binary or Poisson-like outcome analysis is often handled by the generalized estimation equation (GEE) method. Neither method is appropriate for analyzing a multinomial outcome in a longitudinal study, although the cross-sectional multinomial outcome is often analyzed by generalized linear models. One reason that these methods are not used is that the correlation structure of two multinomial variables can not be easily specified. In addition, methods that rely upon GEE or mixed effects models are unsuitable in instances when the focus of a longitudinal study is on the rate of moving from one category to another. In this research, a longitudinal model that has three categories in the outcome variable will be examined. A continuous-time Markov chain model will be used to examine the transition from one category to another. This model permits an unbalanced number of measurements collected on individuals and an uneven duration between pairs of consecutive measurements. In this study, the explicit expression for the transition probability is derived that provides an algebraic form of the likelihood function and hence allows the implementation of the maximum likelihood method. Using this approach, the instantaneous transition rate that is assumed to be a function of the linear combination of independent variables can be estimated. For a comparison between two groups, the odds ratios of occurrence at a particular category and their confidence intervals can be calculated. Empirical studies will be performed to compare the goodness of fit of the proposed method with other available methods. An example will also be used to demonstrate the application of this method. PMID- 16708782 TI - Rethinking prevention. AB - The purpose of this paper was to present information that underscores: (1) the need for understanding the fundamental nature of dental caries; and (2) efforts to minimize its consequences in light of available scientific evidence, recent population trends, and persistent pressures to extend improvements in oral health to all children in the most cost-effective manner. The primary emphasis was placed on the importance of appreciating caries as a common, complex, chronic disease whose deleterious effects can be mitigated best with ongoing use of appropriate, risk-based protective measures. Additional attention was given to current and emerging challenges along with considerations for aligning oral health promotion and delivery system capacity with children's oral health care needs. PMID- 16708783 TI - Examining the cost-effectiveness of early dental visits. AB - The subject of early dental visits as an integral dimension of anticipatory guidance and the related supporting scientific evidence for this concept is a critical and timely issue for the dental profession. The purpose of this paper was to review the scientific evidence and rationale for early dental visits. In theory, early dental visits can prevent disease and reduce costs. During the age 1 dental visit, there is strong emphasis on prevention and parents are given: (1) counseling on infant oral hygiene; (2) home and office-based fluoride therapies; (3) dietary counseling; and (4) information relative to oral habits and dental injury prevention. There is evidence that the early preventive visits can reduce the need for restorative and emergency care, therefore reducing dentally related costs among high-risk children. Preschool Medicaid children who had an early preventive dental visit by age 1 were more likely to use subsequent preventive services and experienced less dentally related costs. These finding have significant policy implications, and more research is needed to examine this effect in a low-risk population. PMID- 16708784 TI - Mutans streptococci: acquisition and transmission. AB - Dental caries is an infectious and transmissible disease. The mutans streptococci (MS) are infectious agents most strongly associated with dental caries. Earlier studies demonstrated that infants acquire MS from their mothers and only after the eruption of primary teeth. More recent studies indicate that MS can colonize the mouths of predentate infants and that horizontal as well as vertical transmission does occur. The purpose of this paper was to demonstrate that these findings will likely facilitate the development of strategies to prevent or delay infant infection by these microbes, thereby reducing the prevalence of dental caries. PMID- 16708785 TI - In defense of the oral cavity: the protective role of the salivary secretions. AB - Saliva performs important protective roles in the oral cavity. Debate in the 1970s over the "specific" or "non-specific" action of salivary components has given way to current attempts to identify the full complement of all proteins in saliva that are now considered to act in concert. At the same time, more fundamental protective qualities of saliva water and pH control are receiving less attention. These qualities may be among saliva's most important. This presentation will review recent advances in the genomics and proteomics of saliva, as well as saliva's roles in tissue coating, alimentation, and regulation of the oral flora. PMID- 16708786 TI - The impact of research and development on the prevention of oral diseases in children and adolescents: an industry perspective. AB - Significant progress has been made globally in reducing dental caries and periodontal disease through adoption of safe and effective methods of prevention. Nonetheless, there are profound oral health disparities in the population and this necessitates further improvement. This review provides an industry perspective of the impact of research and development on the prevention of oral disease in children and adolescents. It also provides an overview and discussion of the key routes that are currently under investigation in academic research, together with an outline of what it will take to develop a major new therapeutic and bring it into clinical practice and individual self care. The key challenges and opportunities are summarized. PMID- 16708787 TI - Caries prevention and reversal based on the caries balance. AB - The science behind caries prevention and reversal is well understood. A recent clinical trial has confirmed that reducing caries risk results in a reduction in dental decay. Dental caries progression or reversal depends upon the balance between demineralization and remineralization and can be visualized for clinical purposes as the "caries balance." This balance is determined by the relative weights of the sums of pathological factors and protective factors. A structured caries risk assessment should be carried out based upon the concept of the caries balance. Following the risk assessment, a treatment plan is devised which leads to the control of dental caries for the patient. The balance between pathological and preventive factors can be swung in the direction of caries intervention and prevention by the active role of the dentist and his/her auxiliary staff. PMID- 16708789 TI - The use of pit and fissure sealants. AB - This paper reviews key issues of sealant use and methodology and poses recommendations to inform the discussion toward a consensus statement by participants. A comprehensive review of sealant literature, including policy recommendations from previous conferences that reviewed best practices for sealant use, was completed. Building on previous review papers and on previous policy statements by dental and public health groups, this paper discusses key questions about sealant use in light of contemporary caries data and cost-benefit analyses. In addition, newest material advancements are reviewed to establish the next step in sealant improvement for young patients. PMID- 16708788 TI - Evidence-based use of fluoride in contemporary pediatric dental practice. AB - Fluoride is an important and effective means of reducing the caries incidence in children. Multiple fluoride products are available to dentists for use with their patients at risk for dental caries. The purposes of this paper are to: (1) review clinically salient evidence, primarily systematic reviews and meta-analyses, for the effectiveness of fluoride options and, where possible, combinations of fluoride exposures; and (2) make recommendations to dental practitioners based on the available evidence for the use of these various approaches in contemporary practice, particularly regarding the use of multiple fluoride sources. The available data suggest that therapeutic use of fluoride for children should focus on regimens that maximize topical contact, preferably in lower-dose, higher frequency approaches. Current best practice includes recommending twice-daily use of a fluoridated dentifrice for children in optimally fluoridated and fluoride deficient communities, coupled with professional application of topical fluoride gel, foam, or varnish. The addition of other fluoride regimens should be based on periodic caries risk assessments, recognizing that the additive effects of multiple fluoride modalities exhibit diminishing returns. PMID- 16708790 TI - A probiotic approach to caries management. AB - The surgical approach has been the predominate mode of caries management for the past 150 years. Dentistry has, however, in recent years moved toward an antibiotic/antimicrobial model of disease management. This approach, however, raises serious questions: (1) do the antibiotic/antimicrobial agents (chlorhexidine, povidone iodine, fluoride, etc) kill all offending organisms?; (2) if so, do the agents preclude the re-entry of the same organisms from external sources?; and (3) if the agents do kill all the offending organisms, do any remaining pathogenic organisms have selective advantage in repopulating the tooth surfaces? To overcome the problems inherent in an antibiotic/antimicrobial approach, probiotic methods are currently under study as means of caries management. This paper discusses probiotic approaches, such as genetically modified Streptococcus mutans and targeted antimicrobials in the management of dental caries. Implications for this approach in the management of other diseases are also presented. PMID- 16708791 TI - Xylitol, sweeteners, and dental caries. AB - The purpose of this report was to provide an overview of xylitol and other polyol sweeteners and dental caries for clinicians and to discuss current applications for dental practice and potential community-based public health interventions. Xylitol, like other polyol sweeteners, is a naturally occurring sugar alcohol. Studies suggest polyols are noncariogenic. Furthermore, studies indicate that xylitol can decrease mutans streptococci levels in plaque and saliva and can reduce dental caries in young children, mothers, and in children via their mothers. Food products containing xylitol are now available and have the potential to be widely accessible to consumers to help control rampant decay. Determining whether products contain adequate xylitol amounts for practical use towards prevention is challenging, however, because xylitol content is not clearly labeled. Sufficient evidence exists to support the use of xylitol to reduce caries. Clinicians and dental associations should push for clear recommendations of efficacious dose and frequency of xylitol use and for clear labeling of xylitol content in products to help consumers choose appropriately. PMID- 16708792 TI - Diet quality, added sugar, and dietary fiber intakes in American preschoolers. AB - PURPOSE: The purpose of this cross-sectional study was to examine the relationship between meeting the intake recommendations for added sugar (AS) and dietary fiber and overall diet quality in American preschoolers from different ethnic backgrounds. METHODS: Data from a nationally representative sample of preschoolers participating in the Continuing Survey of Food Intake by Individuals (CSFII) 1994-1996, and 1998 (n=5,437) were categorized into 3 groups: (1) meeting the recommendation for AS intake (< or = 10% of energy) and dietary fiber (14 g and 20 g in 2- to 3-year-olds and 4- to 5-year-olds, respectively; n=236); (2) meeting only 1 recommendation (n=1,502); and (3) meeting neither (n=3,699). Dietary AS and dietary fiber sources were ascertained. Mean energy intake, nutrient intake, and food group density were established within each category, and student's t test was employed to determine significant differences. Nutrient and food group density was also examined stratified by 4 distinct ethnic groups. RESULTS: Children meeting both recommendations had better diet quality but also higher energy intakes than children meeting only 1 or neither recommendation. CONCLUSION: Most American preschoolers do not meet the dietary intake recommendations for AS and dietary fiber. Health professionals should work together to increase dietary fiber density and reduction of added sugar in the diets of preschoolers to improve diet quality and oral health. PMID- 16708793 TI - Provider versus patient-centered approaches to health promotion with parents of young children: what works/does not work and why. AB - The purpose of this paper was to provide insight into why the health promotion approach now in general use does not work and to describe the rationale and specific steps that can be taken to enhance the effectiveness of preventive communications. PMID- 16708794 TI - Tobacco use by adolescents: the role of the oral health professional in evidence based cessation programs. AB - The use of tobacco products, especially cigarette smoking, represents the leading cause of preventable illness and death in the developed world. In the United States, major gains have been made to reduce smoking among adults. Similar gains, however, have not been realized with adolescents. In recent years, substantial interest has been directed to tobacco cessation studies with adolescents. The previously limited interest in adolescent cessation programs was attributable in large part to the mistaken assumptions that: (1) adolescent tobacco users were not dependent on nicotine and could stop at any time; (2) adolescents did not want to quit; and (3) adult tobacco cessation programs would be effective with adolescents. The need for programs to increase adolescent cessation attempts is underscored by the Healthy People 2010 goal that calls for an increase in tobacco use cessation attempts by adolescent smokers to 84%. Dental providers need to take steps to prevent tobacco use by adolescent patients. For those who are already addicted, they need to provide cessation counseling services or referral for appropriate treatment. The purpose of this paper was to provide dental clinicians with information on: (1) tobacco and health; (2) the epidemiology of adolescent tobacco use; and (3) tobacco cessation programs for parents and adolescents that can be implemented in the dental office setting. PMID- 16708795 TI - Medical approach to dental caries: fight the disease, not the lesion. AB - Dental caries is one of the most prevalent and costly diseases in the United States and throughout the world. Although the manifestation of the disease is the dissolution of tooth structure, the biological nature of the disease is a microbial infection. Traditional dentistry has been heavily focused on repairing damaged tooth structure via surgical approaches. The advancement of microbiology and engineering is bringing a medical revolution to dentistry. The on-going exciting progress in caries research has offered us great opportunities to better understand, detect, and monitor the disease. Most importantly, these new biological discoveries and technological developments allow us to address the etiological cause of the disease (microbial infection), which can lead to more effective treatments and prevention of dental caries. PMID- 16708796 TI - [Expression and location of UBAP1 protein associated with nasopharyngeal carcinoma]. AB - OBJECTIVE: To analyze the expression and location of coding protein of UBAP1 gene and to understand the relationship between the expression pattern of the protein and cell carcinogenesis. METHODS: Bioinformatics was used to analyze the protein character to provide an available clue of subsequent research. The codon frame cDNA was amplified by PCR, and subcloned into enhance green fluorescence protein (EGFP) of pEGFP-C2. The recombinant plasmid was transfected into HNE1 cells. The expression of coding protein was observed by fluorescence microscopy. RESULTS: The expressed GFP-fusion protein generated striking green fluorescence in the cytoplasm in HNE1 cells. EGFP/UBAP1 was expressed and existed mainly in the nuclear, especially accumulated on the nuclear envelope. CONCLUSION: The expression difference in HNE1 might be related to the carcinogenesis of NPC. PMID- 16708798 TI - [Regulation of hepatitis C virus core protein on the activity of signal transducers and activators of transcription 3]. AB - OBJECTIVE: To investigate the regulation of hepatitis C virus (HCV) core protein on the activity of signal transducers and activators of transcription 3 (stat3). METHODS: A cell line expressing stable HCV core protein-QSG7701-core was constructed by transfecting the pcDNA3. 1-core (expressing HCV core protein) into the human immortalized hepatocyte line QSG7701. The phosphorylation and DNA binding activity of stat3 were detected by immunocytochemistry, Western blot, and electrophoretic mobility shift assay (EMSA). RESULTS: The expression level of phosphorylated stat3 in QSG7701-core cells was significantly lower than that in QSG7701-pcDNA3. 1 cells and untransfected QSG7701 cells, but there were no significant differences in the expression levels of total stat3 among the 3 groups. The positive signal of phosphorylated stat3 in nucleus of QSG7701-core cells was obviously weaker than that in QSG7701-pcDNA3. 1 cells and untransfected QSG7701 cells. EMSA showed that DNA binding activity of stat3 in QSG7701-core cells significantly decreased. CONCLUSION: The expressionof HCV core protein in human hepatocyte line may suppress the phosphorylation and DNA binding activity of stat3, which may be one of the causes for resistance against interferon. PMID- 16708797 TI - [Identification of aging related proteins in human normal colonic epithelium]. AB - OBJECTIVE: To explore the molecular mechanisms of colonic epithelial aging related proteins and aged colonic epithelial susceptibility to tumor. METHODS: The proteins of normal human colonic epithelial tissue from young and old people were separated by 2-dimensional gel electrophoresis (2DGE), respectively. Then gels were stained by silver, scanned by imagescanner and analyzed with PDQuest software. The differentially expressed protein spots of colonic epithelium between the old and the young groups were identified by peptide mass fingerprint based on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and database searching. RESULTS: Well-resolved and reproducible 2DGE maps of normal human colonic epithelium from the young and the old were acquired. Nineteen more than 2 fold differentially expressed protein spots were identified representing 17 different proteins by MALDI-TOF-MS. The functions of these proteins involve in metabolism, energy generation, transportation, antioxidation, translation and protein folding. CONCLUSION: Seventeen aging related proteins of human colonic epithelium identified indicate that injury of mitochondrial function and decline of antioxidant capability are important reasons for the aging of human colonic epithelium. These data provided useful clues for elucidating the mechanisms of colonic epithelial aging and aged colonic epithelial susceptibility to cancer. PMID- 16708799 TI - [Construction of two hepatocellular carcinoma cell models for the expression of HBV X gene with different selection characteristics]. AB - OBJECTIVE: To construct 2 hepatocellular carcinoma (HCC) cell models for the expression of HBV X gene with different selection characteristics. METHODS: HepG2 HCC cells were infected with eukaryotic expression vectors with HBV X gene, pCEP4 X, and pcDNA3. 1 (+)-X. Single cell clone was selected by hygromycin and neomycin. After propagating culture for certain periods, the HBV X gene expression was identified by PCR, RT-PCR, and Western blot. RESULTS: Single HCC cell clone with HBV X gene transferred resistant to hygromycin and neomycin was selectively cultured, and the cells could be propagated for certain periods. PCR, RT-PCR, and Western blot identified the expression of HBV X gene. CONCLUSION: Two HCC cell models for the expression of HBV X gene with different selection characteristics have been successfully constructed. PMID- 16708800 TI - [Construction of the eukaryotic expression vector of MJD1 and its expression in SH-SY5Y cells]. AB - OBJECTIVE: To construct the eukaryotic expression vector of MJD1 with normal copies of CAG trinucleotide repetition and MJD1 with CAG trinucleotide repetition expansion mutation respectively, and to determine whether the polyglutamine expansion in ataxin-3 could lead to the formation of intranuclear aggregation. METHODS: The coding sequence of wild-type MJD1 and mutant MJD1 was amplified by PCR from pAS2-1-MJD20Q and pAS2-1-MJD68Q respectively. After being digested with BamH I and Hind III, the PCR products were inserted into pcDNA3. 1-Myc-His(-) B. The recombinant plasmids pcDNA3.1-Myc-His(-) B-MJD20Q and pcDNA3.1-Myc-His(-) B MJD68Q were identified by enzyme digestion analysis and DNA sequencing. The recombinant plasmid was transfected into SH-SYSY cells and the expression of MJD1 in the transfected cells was analyzed by Western blot. The immunofluorescence of the transfected cells was examined using a confocal microscope to observe the formation of intranuclear aggregation. RESULTS: Enzyme digestion analysis and DNA sequencing showed that the target gene was cloned into pcDNA3. 1-Myc-His(-) B. The expression of MJD1 in the transfected cells was confirmed by Western blot; The SH-SY5Y cells transfected with pcDNA3. 1-Myc-His(-) B-MJD68Q showed the formation of intranuclear aggregation, but the cells transfected with pcDNA3.1 Myc-His(-) B-MJD20Q did not show such phenomenon. CONCLUSION: The eukaryotic expression vectors of MJD1 has been successfully constructed; The polyglutamine expansion in ataxin-3 could lead to the formation of intranuclear aggregation. PMID- 16708801 TI - [Effects of vasoactive intestinal peptide on LPS-induced MMP-9 expression by alveolar macrophages in rats]. AB - OBJECTIVE: To explore the role of vasoactive intestinal peptide (VIP) on LPS induced MMP-9 expression by alveolar macrophages (AM) in rats. METHODS: LPS induced cultured Wistar rats AMs were treated with different concentrations of VIP (10(-10) to approximately 10(-6) mol/L) for 24 h. AMs and the supernatant were collected to measure the MMP-9 expression and activity by RT-PCR and gelatin zymography, respectively. Results The MMP-9 activity and expression of LPS induced AMs were significantly higher than those in the control group (P < 0.01). VIP (10(-9) to approximately 10(-6) mol/L) down-regulated LPS-induced MMP-9 activity and its expression. The effects were diminished by H-7 and W-7, an antagonist of protein kinase C (PKC) and calmodulin (CaM) (P < 0.01). CONCLUSION: VIP can decrease LPS-induced MMP-9 activity and its expression, which may be related to protein kinase C and calmodulin pathway. VIP may have protective roles in the lung injury. PMID- 16708802 TI - [Expression of matrix metalloproteinase 2 in oral verruvous carcinoma and squamous cell carcinoma]. AB - OBJECTIVE: To determine the expression of MMP2 mRNA in oral verruvous carcinoma and squamous cell carcinoma. METHODS: Thirty cases were divided into 3 groups: verruvous carcinoma (n = 10), well-differentiated squamous cell carcinoma (n = 15) and moderately or poorly differentiated squamous cell carcinoma (n = 5). Reverse transcription polymerase chain reaction (RT-PCR) was used to test the expression of MMP2 mRNA in the carcinoma tissues and matched normal tissues from 3 groups above. RESULTS: The expression of MMP2 mRNA in the carcinoma tissues was significantly higher than that in their matched normal tissues (P < 0.05). The expression of MMP2 mRNA in verruvous carcinoma was significantly higher than that in well-differentiated and moderately or poorly differentiated squamous cell carcinoma (P < 0.05). However, the expression of MMP2 mRNA was not obviously different between well-differentiated and moderately or poorly differentiated squamous cell carcinoma (P > 0.05). CONCLUSION: The expression of MMP2 mRNA in oral verruvous carcinoma and squamous cell carcinoma tissues was significantly higher than that in their matched normal tissues. The expression of MMP2 mRNA in verruvous carcinoma was significantly higher than that in squamous cell carcinoma. PMID- 16708803 TI - [Confirmation of the extra small chromosome in abnormality karyotype by PCR and FISH]. AB - OBJECTIVE: To investigate the source of the extra small chromosome in a patient with karyotype 45,X[115]/46,X + mar[45]/46,XY[29]. METHODS: The SRYgene was detected by PCR, and the chromosome Y probe that labeled with biotin was detected by fluorescence in situ hybridization. RESULTS: SRY gene is detected positive and the mar chromosome showed positive signal with FISH in human chromosome Y probe pool. CONCLUSION: The extra small chromosome is part of the chromosome Y. PMID- 16708804 TI - [Changes of nerve growth factor and its receptors in the lung tissues in asthmatic rats and their effects on the airway inflammation]. AB - OBJECTIVE: To determine the expression of nerve growth factor (NGF), tyrosine kinase receptor A (trkA), and pan-neurotrophin receptor (p75) in the lung tissues in asthmatic rats, and to explore their effects on the airway inflammation. METHODS: Thirty-two SD rats were randomly divided into 4 groups: the control, asthma, NGF and anti-NGF groups. The asthmatic model was established by the inhalation and injection of ovalbumin. The total cell count and differential cell count in the bronchoalveolar lavage fluid (BALF) were performed. The pathologic changes in the lung tissues of the 4 groups was detected by HE staining. The NGF mRNA expression in the lung tissues of the asthma and control groups was determined by reverse transcription-polymerase chain reaction (RT-PCR). The changes of trkA and p75 mRNA expressions in the lung tissues in the 4 groups were also investigated by RT-PCR. RESULTS: Compared with the control group, the BALF total cell, the BALF eosinophils (Eos), and the BALF lymphocytes (Lyms) significantly increased (All P <0. 001) in the asthma group; and the lung tissues of the asthma group had more infiltrating inflammatory cells. Not only the expression of NGF mRNA, but also its receptors trkA and p75 mRNA in the lung tissues were significantly higher in the asthma group than those in the control group (All P < 0.01). Positive correlation was found between the expression of NGF mRNA and the BALF total cell, the BALF Lyms in the asthma group. Compared with the asthma group, the total cell, the Eos, and the lyms in BALF in the NGF group significantly increased (All P < 0.01), and the lungs of the NGF group had apparent inflammatory changes. The expre-ssions of p75 and trkA mRNA were enhanced significantly (All P < 0.05). Compared with the asthma group, the total cell, the Eos, and the lyms in BALF in the anti-NGF group significantly decreased (All P < 0.001), and the lungs of the anti-NGF group showed alleviative inflammatory changes. The expre-ssions of p75 and trkA mRNA significantly decreased (All P < 0.01). CONCLUSION: In lungs of asthmatic rats, the elevated expression of NGF mRNA is closely related to the airway inflammation. NGF can upregulate the expressions of p75 and trkA mRNA in asthmatic rats, and then may promote their role in the airway neuronal inflammation in asthma. PMID- 16708805 TI - [Effects of nasal continuous positive airway pressure short-term treatment on C reactive protein and intercellular adhesion molecule-1 in patients with obstructive sleep apnea-hypopnea syndrome]. AB - OBJECTIVE: To investgate the effects of nasal continuous positive airway pressure (nC-PAP) short-term treatment on the serum level of C-reactive protein (CRP) and intercellular adhesion molecule-1 in obstructive sleep apnea-hypopnea syndrome (OSAHS) patients. METHODS: Twenty patients with OSAHS ( AHI > or = 15 ) and 15 controls (AHI < 15) were recruited. After polysomnography (PSG), the venous blood was collected from all subjects to investigate CRP and ICAM-1. The effects of short-term treatment (4 d) of nCPAP on the serum levels of CRP and ICAM-1 were studied in patients with moderate and severe OSAHS. RESULTS: Before the treatment, there was a significant positive correlation between CRP and AHI in all subjects (r = 0.615, P < 0.001), a significant negative correlation between CRP and the mean nocturnal oxygen saturation (r = -0.682, P < 0.001), and a significant negative correlation between CRP and the lowest nocturnal SaO2 (r = 0.61, P < 0.001). There was a significant positive correlation between ICAM-1 and AHI in all subjects (r = 0.751, P < 0.001), a significant negative correlation between ICAM-1 and the mean nocturnal oxygen saturation (r = -0.68, P < 0.001), and a significant negative correlation between ICAM-1 and the lowest nocturnal oxygen saturation (r = -0.656, P < 0.001). There was a significant positive correlation between CRP and ICAM-1 (r = 0.437, P = 0.009). The levels of CRP and ICAM-1 were significantly higher in patients with moderate and severe OSAHS than in the controls (P < 0.01). nCPAP decreased the levels of CRP and ICAM-1 in patients with moderate and severe OSAHS (P < 0.01). CONCLUSION: Inflammatory reaction exists in OSAHS and can be palliated after nCPAP short-term treatment. PMID- 16708806 TI - [Protective effects of ulinastatin on the lung injury during cardiopulmonary bypass]. AB - OBJECTIVE: To determine the protective effects and mechanism of ulinastatin on the lung injury during cardiopulmonary bypass (CPB). METHODS: Thirty patients with rheumatic heart disease (RHD) were divided into 2 groups randomly. The ulinastatin group (Group U, n = 15) received 1 x 10(4)U/kg ulinastatin intravenously before the CPB and the same amount of ulinastatin was added into the primary solution. The control group (Group C,n = 15) received normal saline instead of ulinastatin. A brochioalveolar lavage was performed at 2 h after the cardiopulmonary bypass. Polymorphonuclear neutrophil elastase, tumor necrosis factor-alpha and MDA contents in the brochioalveolar lavage fluids were measured, and the lung oxygenate index was measured preoperatively and at 1 and 4 h after CPB termination. RESULTS: Polymorphonuclear neutrophil elastase, tumor necrosis factor-alpha and MDA contents of Group U in the brochioalveolar lavage fulids were significantly lower than those of Group C (P < 0.05), and the lung oxygenate index of Group U at 1 and 4 h after CPB termination was also significantly lower than that of Group C. A significant increase of lung oxygenate index occurred in both groups at 1 and 4 h after CPB when compared with the same group at the baseline before CPB (P < 0.05). CONCLUSION: Ulinastatin has the protective effects on the lung injury during CPB by decreasing polymorphonuclear neutrophil elastase, alleviating lung inflammatory reaction and reducing oxygen free radicals. PMID- 16708807 TI - [Effect of simvastatin on the proliferation of human omental preadipocytes and caspase-3 expression]. AB - OBJECTIVE: To explore the effect of simvastatin on the cell cycle and caspase-3 expression in human omental preadipocytes. METHODS: The preadipocytes were randomly divided into a blank group, a 10(-5) mol/L simvastatin group, and a 10( 4) mol/L simvastatin group. Each group was incubated with different concentrations of simvastatin for 48 hours. MTT method was used to analyze the effect of simvastatin on the proliferation. Distribution of the cell cycle was measured by flow cytometric. Caspase-3 expression was examined by cyto- immunochemistry. RESULTS: After being induced to differentiate for 16 days the human omental preadipocytes developed to mature adipocyte penotypes with lipid droplet. Simvastatin 10(-4) mol/L had significant anti-proliferation effect. Flow cytometric analysis showed the cell cycle was blocked in G0/G1 phase and caspase 3 positive cells increased dramatically. CONCLUSION: Simvastatin may block the cells in G0/G1 phase, and induce caspase-3 expression, which may trigger apoptosis. PMID- 16708808 TI - [Human endostatin gene transfected adult skin melanoma cells]. AB - OBJECTIVE: To study the biological characteristics of human endostatin (hEndo) gene transfected adult skin melanoma cells in vitro and in vivo. METHODS: The plasmid pcDNA3.1 (-)-hEndo was transfected into adult skin melanoma cells by electroporation, and then the stable clones were selected with G418. The transcription and expression of hEndo gene in the transfected melanoma cells were verified by RT-PCR and agarose gel electrophoresis analysis and Western blot. The biological activities of hEndo protein were investigated by MTT in vitro. Stable clones expressing endostatin were subcutaneously injected into the right flank of BALB/c-nu/nu mice of 4 to approximately 6 weeks old. Then the growth of transduced tumors in vivo was investigated. RESULTS: The bands of 624 bp and 5.4 kb were identified from digested plasmid pcDNA3.1 (-)-hEndo. The stable clones were selected with G418 after the eletroporation, the expression of hEndo mRNA was verified by RT-PCR, and Western blot displayed the expression product of hEndo was about 20 kD in the transfected melanoma cells. MTT showed that the conditioned medium of melanoma cells transduced with recombination human endostatin expression vector could inhibit the proliferation of human umbilical vein endothelial cells in vitro. The growth of transduced cells in vivo showed that transfected melanoma cells grew in vivo at a slower rate than the control cells (P < 0.05). RT-PCR showed that endostatin expressed in the transduced tumors. CONCLUSION: Adult skin melanoma cells in vitro transfected with exogenetic hEndo gene can express and secrete active hEndo, and inhibit the growth of transduced tumors in vivo. PMID- 16708809 TI - [Diagnostic value of the detection of aromatase cytochrome P450 and CA125 for endometriosis]. AB - OBJECTIVE: To evaluate the diagnostic value of examining endometrial biopsy specimens for aromatase cytochrome P450 and CA125 for endometriosis. METHODS: The expression of aromatase cytochrome P450 in endometrium was detected by immunohistochemical analysis for 36 endometriosis women and 22 non-endometriosis but subfertility or pelvic pain women served as controls; the serum CA125 levels were measured by chemical radiation method. RESULTS: By measuring serum CA125, the sensitivity of the serum CA125 assay was 44%, the specificity was 82%, the positive predictive value was 80%, and the negative predictive value was 47%; the sensitivity of aromatase cytochrome P450 in biopsy endometrium of endometriosis was 82%, the specificity was 59%, the positive predictive value was 76%, and the negative predictive value was 67%. CONCLUSION: The combination assay of aromatase cytochrome P450 in eutopic endometrium and CA125 can be used as a diagnostic test for endometriosis, especially for the early stage of endometriosis, which is superior to the assay of CA125. PMID- 16708810 TI - [Correlation between dynamic contrast-enhanced MRI and expression of microvascular density in the glioma]. AB - OBJECTIVE: To explore the correlation between microvascular density (MVD) and dynamic contrast-enhanced MRI in the glioma. METHODS: We examined 35 patients with histologically verified glioma. Gadolinium-enhanced dynamic TurboFLASH imaging was performed preoperatively in all patients followed by conventional MRI. The steepest slope (SSmax) of curve and corresponding Tm1 in "first-pass" phase were obtained by analyzing time-signal curve. All specimens were immunostained with anti-human Factor VIII relative antigen monoclonal antibody postoperatively by streptavidin-peroxidase method to obtain the MVD. The correlation between SSmax, Tm1, and MVD was analyzed. RESULTS: SSmax was positively correlated with MVD (r = 0.640, P < 0.01). Tml was negatively correlated with MVD (r = -0.671, P < 0.01). CONCLUSION: The MVD correlates obviously with SSmax and Tml in the glioma. Analyzing the time-signal curve of dynamic contrast-enhanced MRI is helpful to predict the angiogenesis in the glioma. PMID- 16708811 TI - [Clinical observation of young, middle-aged and elderly women with endometrial carcinoma]. AB - OBJECTIVE: To explore the clinical features, diagnosis, treatment and prognosis of endometrial carcinoma in young, middle-aged and elderly women. METHODS: We retrospectively analyzed the clinical data of 82 cases of endometrial carcinoma in young, middle-aged women and 33 cases of endometrial cacinoma in elderly women. RESULTS: The rates of adenocarcinoma in young, middle-aged and elderly groups were 74.4% and 75.5%, respectively. The young,middle-aged and elderly patients with Stage I endometrial cancer were 64.6% and 69.7%, and those with Stage III and IV were 15.9% and 15.2%, respectively. The histological Grade 1 carcinoma of endometrium in young,middle-aged and elderly women were 70.7% and 60.6%, respectively. The young, middle-aged women without myometrial invasion were more than the elderly women (42.8% vs 15.6%, P < 0.01). The young, middle aged women with myometrial invasion more than half of myometrial wall were less than the elderly women (10.4% vs 40.6%, P < 0.01). The rate of chemical treatment after the surgery in the elderly women was more than that of the young, middle aged women (P < 0.05). The 5-year survival rate of the young, middle-aged women was obviously higher than that of the elderly women (92.79% vs 72.21%, P < 0.05). CONCLUSION: Adenocarcinoma and well-differentiated cells are the main pathological characteristics of endometrial carcinoma both in the young, middle aged and the elderly women. Most young, middle-aged and el-derly patients can be diagnosed and treated in the early stage. Early diagnosis and reasonable treatment can improve the prognosis. The prognosis of the young, middle-aged patients is obviously better than that of the elderly patients, and the myometrial invasion depth may be the main difference. PMID- 16708812 TI - [Application of ultrasound-guided percutaneous biopsy in the preoperative diagnosis and treatment of bone tumors]. AB - OBJECTIVE: To investigate the value of color Doppler ultrasonography (CDUS) guided percutaneous biopsy in the diagnosis and treatment of bone tumors. METHODS: Ultrasound-guided percutaneous biopsy was performed in 64 patients who was diagnosed by CDUS. Puncture instruments included automatic core biopsy device and 14 to approximately 18G crude needle. All cases were identified by the operation and pathological examination. RESULTS: Tissue-obtained rate was 97% (62/64). Comparing preoperative pathological diagnosis with postoperative pathological diagnosis, we found the diagnostic correct rate was 97%. No complications resulted from this procedure. According to their pathological results, 33 patients were treated with preoperative chemotherapy. The tumor sizes decreased, and 26 patients saved their limbs and had normal functions. CONCLUSION: CDUS-guided percutaneous biopsy is safe and quick for the preoperative diagnosis and selecting appropriate surgical procedure for bone tumor treatment. PMID- 16708813 TI - [Surgical treatment of thoracic vertebral angioma with extraosseous extension causing spinal cord compression]. AB - OBJECTIVE: To evaluate the therapeutical effect of surgical treatment for thoracic angioma with extraosseous extension which causes spinal cord compression and neurological dysfunction. METHODS: We retrospectively analyzed 5 cases of thoracic vertebral body angioma with extraosseous extension and spinal cord compression. The operation were performed through anterior approach to resect the involved vertebral body and intracanal angioma in 5 patients. Bone graft and internal fixation were also completed in the meantime. Two patients accepted radiotherapy for 2 periods of treatment postoperatively, and the other 3 patients didn't accept the other supportive therapy. RESULTS: The blood loss during the operation was 1000 mL to 1500 mL. All the patients recovered their neurological function rapidly after the operation. The patients were followed up for 1 to 4 years, and no patient recurred. The bone graft was solidly fused in all patients. No hardware failure occurred. CONCLUSION: The surgical treatment for thoracic vertebral body and extraosseous angioma with spinal cord compression is effective, reliable, and workable. Good long-term result can be obtained. PMID- 16708814 TI - [Cervical pedicle screw in the treatment of cervical fracture complicating ankylosing spondylitis]. AB - OBJECTIVE: To determine the effect of cervical pedicle screw in the treatment of cervical fracture complicating ankylosing spondylitis. METHODS: A retrospective study was done in 5 patients of cervical fracture complicating ankylosing spondylitis who were treated with cervical pedicle screw. According to Frankel's standard, 1 patient was in Grade A, 2 patients in Grade B and 1 patient in Grade C and 1 patient in Grade D. All patients were treated with decompression, fusion and posterior fixation with cervical pedicle screw. RESULTS: The average bony healing time was 3.6 months (3.1 to approximately 4.5 months). The instrumentation was not loose, pulled-out or ruptured. Only one C6 pedide was perforated and the vertebral artery was broken, but there was no new postoperative symptom. Three of the patients with neurological deficits improved postoperatively but the other two who were in Grade A were remained unchanged. CONCLUSION: The good biomechanical characteristics of cervical pedicle screw help the successful treatment in the application of cervical fracture complicating ankylosing spondylitis. PMID- 16708815 TI - [Advanced oxidation protein products in diabetic nephropathy patients and its relation with superoxide dismutase, glutathione peroxidase, neopterin]. AB - OBJECTIVE: To determine the changes of advanced oxidation protein products (AOPP) in diabetic nephropathy (DN) patients, as well as its relationship with superoxide dismutase (SOD), glutathione peroxidase (GPx) and neopterin (NPT). METHODS: By the concentration of urinary albumin excretion rate (UAER) and creatinine (Cr), 85 diabetes mellitus (DM) patients were divided into 4 groups as non-DN group (DM), early-staged DN group (DN3), significant DN group (DN4) and end-staged DN group (DN5). The concentration of the serum AOPP was measured by ameliorated method introduced by Wikto-Sarsat, while SOD by Xanthine oxidase test, GPx by [5,5'Dithio-bis (2-Nitrobenzoic aicd) ] (DTNB) reaction test and NPT by ELISA. RESULTS: AOPP in Group DN5 [(117.8 +/- 64.8) [micromol/L] and Group DN4 [ (80.0 +/- 23.0) micromol/L] were significantly higher than those in Group DM [(58.2 +/- 17.7) micromol/L]. There was no significant difference of AOPP between Group DN3 [(72.7 +/-17.2) micromol/L] and Group DM. Serum AOPP negatively correlated with SOD and GPx (r = -0.217 and -0.374 respectively, P < 0.05), while positively correlated with NPT (r = 0.499, P < 0.01). CONCLUSION: DN patient has enhanced protein oxidation than DM patient, which is related to oxidative stress and chronic inflammation in DN. PMID- 16708817 TI - [Effect of nutrient support on severe infant pneumonia]. AB - OBJECTIVE: To explore the effect of nutrient support on severe infant pneumonia. METHODS: Prospective study was conducted on the outcome of 567 inpatients suffering from severe pneumonia in 13 hospitals randomly selected in Hunan. Twelve factors were surveyed and data analyzed by multiple logistic regression. RESULTS: Malnnutrition, anemia and rickets were risk factors in severe pneumonia, and nutrient support had protective effect on severe pneumonia. CONCLUSION: Nutrient support contributes to the positive outcome of severe infant pneumonia. PMID- 16708816 TI - [Therapeutic effect of harnal and proscar in treating benign prostatic hyperplasia]. AB - OBJECTIVE: To evaluate the therapeutic effect of harnal and proscar in treating benign prostatic hyperplasia (BPH). METHODS: Two hundreds and twenty-two patients with BPH were randomly assigned into 2 groups:harnal (0.2 mg/d) group (n = 112) and proscar (5 mg/d) group (n = 108). American Urologic Association Symptom Index (AUA-SI) scores, the maximal urinary flow rate (Qmax) and prostatic volume were analyzed in the 2 groups. RESULTS: After 12 weeks of treatment, 54.5% of the harnal group improved in AUA-SI score; 54.6% of the proscar group improved in AUA SI score; and there was no significant difference between the 2 groups (P > 0.05). After 24 weeks of treatment, the proscar group (79.6%) had a greater improvement in AUA-SI score than the harnal group (64.3%) (P < 0.05); Qmax in the 2 groups was significantly increased than that before the treatment, and it is proportional to the therapeutic time. The prostatic volume in 2 groups had no significant change. CONCLUSION: Harnal and proscar can significantly improve BPH symptoms and the urinary flow rate. The therapeutic effect of harnal and proscar has their own characteristics. PMID- 16708818 TI - [Therapeutic effects of the combination of traditional Chinese medicine and western medicine on patients with peptic ulcers]. AB - OBJECTIVE: To explore the therapeutic effects and mechanisms of the combination of traditional Chinese medicine and western medicine on patients with peptic ulcers. METHODS: One hundred and twenty patients were randomly divided into 6 groups. Another 10 patients as the control group were confirmed with no peptic ulcers by endoscope, but had digestive tract symptoms. The clinical effects were compared among each group after the one month treatment. RESULTS: The clinical effects of the combination of Jianweiyuyang granules and ranitidine capsules were better than those of western medicine, with improvement in symptoms and syndrome (P < 0.01 to 0.05), but there was not significant difference with the rate of ulcer healing and the Hp clearance among the combination of Jianweiyuyang granules and ranitidine capsules, Jianweiyuyang granules, and ranitidine capsules (P > 0.05). The combination of Jianweiyuyang granules and ranitidine capsules could significantly upregulate the expression of MUCSAC mRNA (P < 0.01), while downregulate the expression of ETAR mRNA (P < 0.01). CONCLUSION: There is obvious advantage in treating peptic ulcers by the combination of Jianweiyuyang granules and ranitidine capsules, and its mechanisms may be to protect the gastric mucosal barrier by up-regulating the expression of MUCSAC mRNA and to improve the gastric mucosal blood flow by down-regulating the expression of ETAR mRNA. PMID- 16708820 TI - [Progress of the immune escape mechanisms of chlamydiae]. PMID- 16708819 TI - [Antihypertensive therapy in 357 inpatients of elderly isolated systolic hypertension]. AB - OBJECTIVE: To determine the therapeutic status and rate of reaching target blood pressure in elderly isolated systolic hypertension (EISH) patients who were hospitalized and to discuss the rationality of the drug therapy. METHODS: Three hundred and fifty-seven EISH inpatients were investigated retrospectively. The frequencies of using antihypertensive drugs and the strategy of drug therapy programs were calculated. The drug efficacies were assessed among various drug therapy groups. RESULTS: The frequencies of using antihypertensive medicine categories were calcium channel blocker (CCB) 64. 15%, angiotensin-converting enzyme inhibitor (ACEI) 32.77%, diuretics 26.33%, beta-blocker (BB) 25.77%, angiotensin receptor blocker (ARB) 23.81%, and alpha-blocker 4.20%, respectively. Among the 357 cases 42.86% were treated with monotherapy while 57.14% with combined therapy. Among the combination therapy groups, the diuretic-based multiple therapy occupied 16.53%, and the non-diuretic-based multiple therapy held 40.62%. The systolic blood pressure control rate was 67.79%. The rate of diastolic blood pressure < or = 70 mmHg was 26.89%, 8 cases of them occurred myocardial ischemia. CONCLUSION: How to select the optimal antihypertensive drug therapy for EISH patients is important in achieving the blood pressure goal. While thinking over intervention at lower blood pressure levels to achieve target goals, physicians should prevent from excessively lowering the diastolic blood pressure. PMID- 16708822 TI - [Cathepsin B expression and its significance in non-small cell lung cancer tissues]. PMID- 16708821 TI - [Stimulation of bone marrow fibroblst conditioned medium on megakaryocyte in vitro and platelet in vivo]. PMID- 16708823 TI - [Measurements of interleukin-18 in peritoneal fluid and serum of patients with endometriosis]. PMID- 16708824 TI - [Analysis of clinical data and pathological types in 1 211 adult patients with renal biopsy]. PMID- 16708825 TI - [Application of percutaneous dilatational tracheostomy in severe brain injury]. PMID- 16708826 TI - [A case of intracranial multiple aneurysms secondary to left atrial myxoma]. PMID- 16708827 TI - Individual differences: signal, noise--or both? PMID- 16708828 TI - Increased pain sensitivity to intraoral capsaicin in patients with atypical odontalgia. AB - AIMS: To use 2 well-characterized stimuli, the intraoral capsaicin model and the "nociceptive-specific" electrode, to compare superficial nociceptive function between patients with atypical odontalgia (AO) and matched healthy controls. Furthermore, the authors aimed to describe the sensitivity, specificity, and positive predictive values (PPV) of the techniques if group differences could be established. METHODS: Thirty-eight patients with AO and 27 matched healthy controls participated in this study. Thirty microliters of 5% capsaicin was applied to the gingiva on the left and right sides of all participants as a pain provocation test. The participants scored the capsaicin-evoked pain continuously on a 0-to-10 visual analog scale (VAS). Furthermore, individual electrical sensory and pain thresholds to stimulation with a "nociceptive-specific" electrode on the facial skin above the infraorbital or mental nerve were determined. RESULTS: AO patients had higher VAS pain scores for capsaicin application than healthy controls (ANOVA: F > 4.88; P < .029). No differences between the painful sides and the nonpainful sides of the patients were found (ANOVA: F < 1.26; P > .262). No main effects of group or stimulation side on the electrical sensory and pain thresholds were detected (ANOVA: F < 0.309; P > .579). Sensitivity was 0.51; specificity, 0.81; and PPV, 0.77 when a VAS value of > or = 8 for capsaicin-evoked pain was used. CONCLUSION: AO patients show increased sensitivity to intraoral capsaicin but normal sensitivity to "nociceptive-specific" electrical stimulation of the face in an area proximal to the painful site. The use of the intraoral pain-provocation test with capsaicin as a possible adjunct to the diagnostic workup is hampered by the only moderately good sensitivity and specificity. PMID- 16708829 TI - Self-efficacy is associated with pain, functioning, and coping in patients with chronic temporomandibular disorder pain. AB - AIMS: To examine the psychometric characteristics of a measure of self-efficacy for managing temporomandibular disorders (TMD) and to determine whether scores on this measure were related to pain, disability, and psychological distress in patients with chronic TMD pain. METHODS: Patients seeking treatment for chronic TMD pain (n = 156, 87% female, mean age = 37 years) completed measures assessing pain, disability, mental health, pain-coping strategies, and self-efficacy for managing their pain. RESULTS: The self-efficacy measure, which was adapted from arthritis research, demonstrated good psychometric characteristics (Cronbach's alpha = 0.91, minimal floor and ceiling effects, and validity). Greater self efficacy was associated with significantly (P < .05) lower levels of pain, disability, and psychological distress. Self-efficacy remained significantly associated with disability and mental health measures even after controlling for demographic variables and pain intensity. In addition, patients with higher self efficacy reported significantly (P < .05) greater use of an active, adaptive chronic pain-coping strategy (task persistence) and less use of a passive, maladaptive chronic pain-coping strategy (rest). CONCLUSION: Self-efficacy for managing pain appears to be important in the adjustment of patients with chronic TMD pain. Research is needed to determine whether treatments designed to increase self-efficacy improve TMD patient outcomes. PMID- 16708830 TI - Daily pain coping among patients with chronic temporomandibular disorder pain: an electronic diary study. AB - AIMS: To describe patients' daily coping with the pain of chronic temporomandibular disorders (TMD), the conservative treatment received, and the self-care strategies used, and to examine the relationships between these strategies and daily pain intensity, activity interference, and jaw use limitations. METHODS: TMD clinic patients (n = 137, 88% women) completed electronic diary measures of pain, interference, jaw use limitations, and use of 20 strategies 3 times daily for 2 weeks. RESULTS: Reliability and validity were demonstrated for 4 scales of related coping items: cognitive coping, relaxation, activity reduction, and emotional support. Average scores were higher on the relaxation and activity reduction scales than on the cognitive coping and emotional support scales. Among the coping items not included in the scales, "did something to try to reduce pain" (direct action) was endorsed most frequently (reported in a median of 74% of interviews). Heat, cold, and seeking spiritual support were used least (< or = 5%). At times of increased pain, patients were more likely to use almost all types of strategies. Pain intensity was associated strongly with concurrent activity interference and jaw use limitations. When the design controlled for pain intensity, activity reduction and seeking emotional support were associated positively within-subjects with interference and jaw use limitations. CONCLUSION: TMD clinic patients use a variety of treatment, self care, and coping strategies to contend with daily pain. Inquiring about a broad range of strategies might help clinicians better understand how individual patients approach pain management. Research is needed to examine whether decreasing activity reduction and emotional support coping results in improved outcomes. PMID- 16708831 TI - The reliability and validity of self-reported temporomandibular disorder pain in adolescents. AB - AIMS: To evaluate the reliability and validity of self-reported pain associated with temporomandibular disorders (TMD) in adolescents and to determine how this validity may change over time. The authors' hypothesis was that self-reported pain can be used to reliably and accurately detect adolescents with TMD pain. METHODS: One hundred twenty adolescents, 60 with self-reported TMD pain and 60 age- and gender-matched controls without TMD pain, were examined twice. At the first examination at a Public Dental Service clinic, self-reported TMD pain was recorded for each patient. At the second examination, a clinical examination was completed, blind to the patients' self-report of pain symptoms, after which self reported TMD pain was again recorded. The clinical examination was based upon the Research Diagnostic Criteria for TMD (RDC/TMD). Self-reported TMD pain in this investigation was based upon the subjects' responses to 2 questions: (1) Do you have pain in your temples, face, temporomandibular joint (TMJ), or jaws once a week or more? and (2) Do you have pain when you open your mouth wide or chew once a week or more? RESULTS: Test-retest reliability of .83 (kappa) was found for the 2 questions. The sensitivity was .98 (95% CI, .90 to 1.0) and specificity was .90 (95% CI, .81 to .95) for comparison of assessments made on the same day. Sensitivity was .96 (95% CI, .85 to .99) and specificity .83 (95% CI, .72 to .90) for assessments made 2 to 4 weeks apart. CONCLUSION: Very good reliability and high validity were found for the self-reported pain questions. A short time interval between the screening question and examination slightly increased the accuracy of the measure. In adolescent populations, the questions in this study can be used to screen for TMD pain. PMID- 16708832 TI - The excitability of the trigeminal motor system in sleep bruxism: a transcranial magnetic stimulation and brainstem reflex study. AB - AIMS: Since sleep bruxism (SB) is characterized by grinding and clenching of the teeth during sleep and could be an exaggerated manifestation of normal spontaneous rhythmic masticatory muscle activity, the aim of this study was to obtain a neurophysiological assessment of the excitability of the central jaw motor pathways in patients with signs and symptoms suggestive of SB. METHODS: A total of 30 subjects diagnosed with SB on the basis of self-report of tooth grinding were studied using the "recovery cycle" of the masseter inhibitory reflex (MIR) elicited by electric and magnetic stimulation of the mental nerves and by recording the motor potentials evoked in masseter muscles by transcranial magnetic stimulation. Tests were done during daytime, when the subjects were awake. The data obtained were compared with data from a population of normal subjects. RESULTS: In the putative SB patients and in normal subjects, the MIRs evoked by single electric and magnetic stimuli were similar. With paired stimuli, the degree of suppression of the late silent period was significantly lower (P < .01) in the patients compared to normal subjects, particularly for magnetic stimuli, at various interstimulus intervals. No significant differences were found between the 2 groups of subjects in the masseter motor potentials evoked by transcranial magnetic stimulation. CONCLUSION: Although the data were only obtained during wakefulness in patients self-reporting signs and symptoms suggestive of SB, the findings suggest that an abnormal excitability of the central jaw motor pathways may be present in SB subjects. This increased excitability could derive from an impaired modulation of brainstem inhibitory circuits and not from altered cortical mechanisms. These results support the view that bruxism is mainly centrally mediated and that it involves subcortical structures. The study also indicates that use of the MIR elicited by the double shock technique could be valuable in the evaluation of bruxism. PMID- 16708833 TI - The effects of age and sex on the expression of aromatase in the rat temporomandibular joint. AB - AIMS: To investigate the expression of aromatase in temporomandibular joints(TMJs) of male and female rats of different ages. METHODS: The expression of aromatase in terms of protein and mRNA was examined by immunohistochemistry, Western blot, and in situ hybridization in the TMJs of Sprague-Dawley rats. The expression level of aromatase protein was also quantified by the density of aromatase-positive chondrocytes in TMJ condylar cartilage. A tritiated water assay was used to analyze the activity of aromatase in cartilage chondrocytes. RESULTS: Strong aromatase protein and mRNA hybridization signals were detected in hypertrophic and mature chondrocytes in all the cases examined. The density of aromatase-positive chondrocytes was relatively stable at higher levels before 8 weeks of age. The density decreased gradually in females after 8 weeks of age but not in males. CONCLUSION: These results demonstrate that aromatase is expressed intensely in condylar cartilage and suggest that it may play an important role in pathophysiological mechanisms in condylar cartilage. PMID- 16708834 TI - An animal model for inducing anterior disc displacement of the temporomandibular joint. AB - AIMS: To develop an animal model of anterior disc displacement (ADD) without the need for opening the temporomandibular joint (TMI) capsule. METHODS: Thirty-two healthy adult Japanese white rabbits were used in this study. Four rabbits were dissected to familiarize the investigators with the anatomy of the TMJ. Sixteen animals were subjected to surgical ADD in the right TMJ, 8 animals had a sham operation, and 4 animals were not operated (normal controls). Four rabbits from the experimental ADD group and 2 from the sham group were sacrificed 1, 2, 4, and 8 weeks, respectively. The rabbits in normal control group were sacrificed at the beginning of the experiment. Animal behaviors as well as macro- and microchanges in the TMJs were investigated. RESULTS: Fifteen right TMJ discs in the 16 experimental rabbits were successfully displaced anteriorly, and the degree of ADD in the experimental group was similar. The mandible of each ADD rabbit deviated to the left side with inclined attrition of the incisors. Some histologic changes appeared in the experimental TMJs. CONCLUSION: This ADD technique without the need for opening the TMJ capsule is effective, and the model is suitable for studying ADD of the TMJ. PMID- 16708835 TI - 4th International Conference on Orofacial Pain and Temporomandibular Disorders, August 26-28, 2005, Sydney, Australia. PMID- 16708836 TI - [The influence of mesenchymal stem cells on bone tissue regeneration upon implantation of demineralized bone matrix]. AB - Mesenchymal stem cells (MSC) are resident pluripotent cells of bone marrow stroma. MSC are able to differentiate into chondroblasts, adipocytes, neurons, glia, cardiomyocytes, or osteoblasts. The problem of MSC usage in cell therapy of bone defects is widely discussed at present. The experiments were carried out using rats of inbred line Wistar-Kyoto. MSC were isolated from bone marrow and cultivated in vitro. Demineralized bone matrices (DBM) were obtained from parietal bones of rats and hens. Part of DBM was loaded with MSC. Bone defects were made in cranium parietal regions. DBM with or without MSC or metal plates were transplanted in these regions. It was shown that the application of MSC increased angiogenesis and osteogenesis in the damaged bone. The implantation of rat's DBM with MSC led to the formation of a full value bone. MSC suppressed inflammation, when transplantation of hen's DBM was carried out. The application of MSC always improved bone tissue regeneration. PMID- 16708837 TI - A primary culture of haemocytes isolated from Gryllus bimaculatus (Orthoptera, Gryllidae) and their interactions with two intracellular parasites--Paranosema grylli (Microsporidia) and Adelina grylli (Coccidia). AB - Cricket haemocytes were derived from either haemolymph or haemopoietic organs (lymph glands) of insects and introduced to a primary culture. Varied isolation protocols, tissue culture vessels, media compositions and cell densities were tested to determine the optimal conditions for in vitro maintenance of haemocytes, and for subsequent light and electron microscopic analysis of monolayers. Freshly prepared Mitsuhashi and Maramorosh (MM;Sigma, Steinheim, Germany) insect medium (420 mOsm), buffered with sodium bicarbonate (pH 7.2) and supplemented with 10 % FCS, was found to be most appropriate for haemocyte maintenance. All tested tissue culture vessels (FLEXiperm units, multiwell plates and Thermanox slides, with the exception of Melineux agar plates), were suitable for cell attachment and haemocyte monolayers formation. Viability of cultured cells was confirmed by LIVE/DEAD Viability/Cytotoxity Kit for Eukaryotic Cells. Free circulating haemocytes were cultivated up to 27 days and then degraded. Infection with the microsporidian Paranosema grylli or the coccidian Adelina grylli caused noticeable swelling of host lymph glands (haemopoietic tissue) and increase in the number of cells comprising the glands. The cells derived from haemopoietic tissue were maintained for maximum 5 days; thereafter multiplication of bacteria normally inhabiting cricket lymph glands destroyed monolayers and killed the cells. Microsporidian and coccidian invasive stages (spores and sporozoites, respectively) were isolated from infected tissues, resuspended in MM medium and added to haemocyte monolayers in ratios 1 zoite per haemocytes or 10 spores per 1 haemocyte. Actively moving zoites contacted and penetrated the cultured cells. Unlike coccidian zoites, microsporidian spores were phagocytized by haemocytes. Application of fluorescent LIVE/DEAD kit allowed to visualize internalized parasites inside host cells as clearly shaped dark areas. The present study has demonstrated that 1) cricket haemocytes from both circulating haemolymph and lymph glands can be short-term cultivated on tissue culture vessel surfaces which made possible their further light and electron microscopic analysis; 2) short-term haemocyte cultures may be employed to study host-parasite interactions, in particular, to follow the initial steps of parasite internalization inside host cell; 3) Fluorescent assay with Viability/Cytotoxity Kit for Eukaryotic Cells (Molecular Probes, Oregon) allows to observe penetration of these parasites into cultured cells. PMID- 16708838 TI - [Ultrastructural characteristics of the reticular thalamic nucleus neurons in WAG/Rij rats]. AB - The aim of this study was to investigate the ultrastructure of the reticular thalamic nucleus (RTN) in rats of WAG/Rij strain, an established model for human absence epilepsy. Most RTN neurons are medium-to large-sized and have either dark or light appearance, depending on their functional state. Moreover, small-sized neurons with short axons are present, their characteristics being described for the first time. PMID- 16708839 TI - Effect of lactoferrin on the viability of Arn8 and HaCaT cell lines. AB - The results on the effect of LF from different sources on the viability of two cell lines (Arn8 and HaCaT) are presented in the work. It is shown that both hLF and bLF have an effect on the viability of these two cell lines, and that bLF decreases the cell viability in much more extend than hLF. Possible action mechanisms of LF are discussed. PMID- 16708840 TI - [Comparative study of the influence of renal endogenic factors on the proliferation activity of epithelial cells]. AB - The proliferation activity of monolayer culture of Madin Darby Canine Kidney (MDSK) cells is suppressed by a thermostable protein factor of renal tissue of white rats and of humans. Under the influence of renal factors (RF), a decrease in cell number, and suppression of DNA synthesis and mitotic activity in MDCK cells occur. The inhibition of proliferative activity of cultured cells under the influence of RF was substantiated also by MTT assay. It was established that the inhibitory influence of RF is stipulated by suppression of RNA synthesis. It follows that RF may inhibit division of MDCK cells via suppression of gene expression in G1-phase. Similar factors were obtained from renal cells of different systematic groups of organisms (snail, frog, fish, pigeon, guinea pig, swine). PMID- 16708841 TI - [Kinetic features of a white rat hepatocyte population under hormonal disbalance before and after partial hepatectomy]. AB - Bilateral adrenalectomy, followed in 4 days by a partial hepatectomy, was performed using white rats weighing as much as 120-140 g. Under hormonal disbalance caused by bilateral adrenalectomy, the number of polyploid (4c, 4c x 2, and 8c) hepatocytes significantly increased, compared to that in non-operated control rats. Six hours after a partial hepatectomy, the share of highly ploid hepatocytes falls, being accompanied by a 9-fold increase in mitotic index. It is supposed that under hormonal disbalance condition, a partial hepatectomy may induce "early" mitoses in hepatocytes blocked in G2-phase of the cell cycle. PMID- 16708843 TI - [Electron dense substance of thylakoids in chloroplasts of tansy Tanacetum vulgare L]. AB - An intrathylakoid electron opaque substance, further referred to as loculin, is found in 80-90 % of thylakoids of tansy leaf mesophyll chloroplasts at the stage of flower bud formation and flowering. Upon conventional isolation of chloroplasts in aqueous solution, and fixation in osmium solution alone, loculin is not retained in thylakoids. Preliminary fixation of leaves in glutaraldehyde makes it possible to isolate chloroplasts without injuring the envelope and stroma (glutar chloroplasts), and loculin is retained in thylakoids under these conditions. Upon prolonged incubation of glutar chloroplasts (for 24 h), loculin leaves thylakoids in the form of drops concentrating on the chloroplast envelope. Upon crossing the thylakoid membrane and chloroplast, loculin properties remain unchanged. It is assumed that loculin is an important metabolite necessary for active growth. PMID- 16708842 TI - [Induction of meiotic disturbances in spermatocytes I by pheromones as an inhibiting mechanism of male reproductive function in house mice]. AB - The influence of pheromons on reproduction and other important physiological characteristics has been reported for many mammalian species. However, mechanisms of this action at the level of target cells still remain unclear. A study was made of the influence of non-identified pheromones from adult males and a female pheromone 2,5-dimethylpyrazine on germ cells of CBA inbred strain mice. Cytogenetic analysis shows a significant increase in such meiotic disturbances as multivalent associations and autsomal univalents 24 h after exposure to pheromonal cues. Results of in situ hybridization show that the level of c-fos and c-jun expression is significantly higher 3.5 h after exposure to pheromones of adult males. It is likely that destabilization of chromosomal apparatus in dividing meiotic cells forms the basis of some reproductive effects of murine pheromones. Possible mechanisms of pheromone influence on reproduction are discussed. PMID- 16708844 TI - [Temperature impact on localization of "free" phenolic compounds in the root tissues and deformation of root hairs in pea seedlings inoculated by Rhizobium]. AB - The study was focused on localization of "free" phenolic compounds in pea Pisum sativum L. seedling roots grown at 22 and 8 degrees C 24 h after their inoculation with Rhizobium leguminosarum bv. viceae bacteria. A comparison of phenolic compound distribution along the root in root tissues, and results of observation of root hair development on the root surface, response of root hairs to inoculation, manifesting itself in various deformation degree (bends, twists, ect.) enabled us to reveal differences between roots grown at different temperatures. These differences are basically referred to a sector localized 0-5 mm away from the root tip containing meristematic and extending cells. A distribution of phenolic compounds in sectors with root hairs responding to inoculation by various degrees of contortion practically did not depend on the temperature of plant growth. The evidence provided in the course of this work enabled us to suggest that inhibition of pea root infection at low temperature is caused by decelerated growth processes characteristic of both the root itself and root hairs, as well by a slow increase in the root hair zone. PMID- 16708845 TI - [Scoring criteria of aneuploidy frequency in interphase nuclei by fish-analysis]. AB - Principles and approaches of assessment of numerical chromosome abnormalities in interphase nuclei by fluorescent in situ hybridization technology are reviewed. The authors' own scoring criteria of results of hybridization of centromere specific DNA-probes with chromosomal targets in somatic cells by dual color FISH analysis are suggested. Use of these scoring criteria allows to reduce the level of artificial hypoploidy, whose its frequency becomes similar to that of hyperploidy. This fact indicates that chromosomal nondisjunction is a major mechanism of aneuploidy induction, rather than of chromosome loss. PMID- 16708846 TI - [Multifunctional nuclear protein NAP57 specifically interacts with dead RNA helicase p68]. AB - NAP57 has been found as a component of nuclear matrix protein complex with ability to specifically bind alphoid DNA. Polyclonal antibodies against NAP57 were raised in order to investigate intranuclear localization and interactions of the protein. Two types of localization were observed: a) nucleoplasmic and b) nucleolar. A bulk of nucleoplasmic fraction is present in splicing factors compartments (SFC). The type of localization pattern does not depend on the cell cycle phase, but we revealed changes in NAP57 localization pattern during S phase. According to immunoprecipitation and immunofluorescence assays, NAP57 specifically interacts with DEAD RNA helicase p68 in vitro and co-localizes with helicase p68 in the nucleus of cultured cells. We suppose participation of both proteins in processing of small nuclear RNA on the SFC periphery, and positioning of the nucleolus according to centromere regions of chromosomes. PMID- 16708847 TI - [Molecular causes of changes in sensitivity of adenylyl cyclase signaling system to biogenic amines in the heart muscle during experimental streptozotocin diabetes]. AB - Changes in hormonal sensitivity of the adenylyl cyclase signaling system (ACS) and their possible molecular causes in the heart muscle of rats with experimental streptozotocin diabetes (type I diabetes) are investigated. An increase in stimulating effects of noradrenaline and isoproterenol on adenylyl cyclase (AC) activity have been shown. In the case of noradrenaline, this increase is due to suppression of Gi-protein function and Gi-coupled inhibitory AC signaling pathway. Meanwhile, in diabetic rats the influence of C-terminal peptide 346-355 of alphai2-subunit on hormonal activation of AC and GTP-binding is diminished. In the case of isoproterenol, along with its stimulating effect, at micromolar concentrations this hormone exerts inhibitory action, realized, presu- mably, through beta3-adrenergic receptors. Effect of isoproterenol on AC and GTP-binding in the heart of diabetic animals is modified by peptide 385-394 alphas, blocking Gs-coupled signaling pathways, and by peptide 346-355 alphai2, blocking transduction of inhibitory signals. In addition, a decrease in serotonin stimulating effect on components of ACS in diabetic animals was shown. The data obtained provide evidence for changes in ACS function in diabetes, which can be detected mainly at the G-protein level. The proposed peptide strategy is a new and perspective approach for studying molecular causes of functional violations in hormonal signaling systems arising at endocrine pathology. PMID- 16708848 TI - [Nitroblue tetrazolium reduction by human blood neutrophils. I. The influence of pH]. AB - Activity of blood neutrophils of 14 healthy volunteers (8 women, 6 men, aged from 21 to 37 years) was investigated in the Nitroblue Tetrazolium (NBT) reduction reaction at pH 7.0, 7.3, and 7.6. The dependence of the percentage of NBT positive neutrophils on the reaction time was described by a kinetic equation with constants k1 and k2, characterizing, accordingly, speeds of NBT absorption and reduction by neutrophils. A strong positive correlation (r = 0.99; P < 0.0001), was found between k1 and k2, which pointed to dependence of the speed of NBT restoring by neutrophils on the speed of its entry to the cell. Both the constants and speeds of successive process stages have maximum values at pH 7.3 (corresponding to physiological value of blood pH), and decrease at medium acidification or alkalization. The k1/k2 ratio does not depend on the time (0-6 h) or pH (7.0-7.6) of blood incubation before the reaction. The most active neutrophils (potential pH agocytes) have a lesser membrane permeability than do neutrophils of another class, and their activity more increases at medium acidification in vitro and, probably, in vivo. The activation of neutrophils leads to an increase in medium pH that alongside with an increase in the number of active potential phagocytes, evidently has a physiological meaning. PMID- 16708849 TI - [Nitroblue tetrazolium reduction by human blood neutrophils. II. The influence of sodium and potassium ions]. AB - Activity of blood neutrophils of 18 healthy volunteers (4 men and 14 women aged from 20 to 32 years) was investigated in the Nitroblue Tetrazolium (NBT) reduction reaction in Na- and K-phosphate buffers (0.1 M, pH 7.3) after a preliminary incubation of heparinized blood (37 degrees C, 0-7 h). It was found that the percentage of NBT-positive neutrophils in the presence of Na+ is higher than in the presence of K+. The dependence of percentage of NBT-positive neutrophils on the reaction time is described by a kinetic equation with constants k1 and k2, characterizing accordingly, speeds of NBT absorption and reduction by neutrophils. It has been shown that k1, k2 and k1/k2 values differ in potential phagocytes and in neutrophils, which are not capable of adherence. The kinetics constants of the latter cells do not depend on both cations and incubation time, while the percentage of NBT-positive potential phagocytes, and their k1 and k2 in the presence of Na+ are higher than in the presence of K+, and the values of described parameters are changed during incubation. During incubation the percentage of NBT-positive neutrophils increases to result in a partial aggregation of thrombocytes, as a consequence of excretion in the blood of products involved in this process. PMID- 16708850 TI - A retrospective long term study of teeth restored with prefabricated carbon fiber reinforced epoxy resin posts. AB - The Composipost endodontic post, made of stretched aligned carbon fibers embedded in an epoxy-resin matrix, has since the beginning of the nineties been widely used for the restoration of endodontically treated teeth. The aim of this retrospective study was to evaluate the treatment outcome of the Composipost system up to 7 years. In a study published 1998, 236 patients treated during 1992 93 by seven Swedish general dental private practitioners were studied. Five of the former seven private practitionersi consented to participate in this follow up of that study. Thus the material was reduced to 138 patients. Thirty-nine of these were excluded due to insufficient data. For the remaining 99 patients, data were collected from dental records. All patients were offered a clinical examination but only 25 accepted. Data were collected from dental records for the remaining 74 patients. The mean follow up time was 6,7 years with a range from 1 month to 10 years (median 7.6 years, SD 2.5 years), (five teeth were extracted during the previous study). The outcome was considered successful if the post and core was in situ and showed no clinical or radiographic signs of technical failures. Sixty-four teeth (65%) restored with the Composipost system were successful after a mean time of 6.7 years. Thirty-two teeth were extracted due to fractures, periapical lesions and periodontitis. Dislodgment of post was observed in three cases. In conclusion, within the limitations of this study, after a mean time of 6.7 years, the Composipost restored teeth had shorter survival times than those of previously documented cast posts. PMID- 16708851 TI - Impaired positioning of the gape in whiplash-associated disorders. AB - We have previously introduced a new concept for natural jaw function suggesting that "functional jaw movements" are the result of coordinated jaw and neck muscle activation, leading to simultaneous movements in the temporomandibular, atlanto occipital and cervical spine joints. Thus, jaw function requires a healthy state of both the jaw and the neck motor systems. The aim of this study was to examine the positioning of the gape in space during maximal jaw opening at fast and slow speed in healthy as well as whiplash-associated disorders (WAD) individuals. A wireless optoelectronic technique for three-dimensional movement recording was used. Subjects were seated in an upright position, with back support up to the mid-scapular level without headrest. The position of the gape in space was defined as the vertical midpoint position of the gape at maximal jaw opening (MP). In healthy, the MP generally coincided with the reference position at the start of jaw opening. In the WAD group, the MP was significantly lower than the reference position. No sex or speed related differences were found. The results suggest that both the width and orientation of the gape in space relies on coordinated jaw and neck muscle activation and mandibular and head-neck movements. This study also suggests an association between neck pain and dysfunction following trauma, and reduced width and impaired positioning of the gape in space. Finally, the MP seems to be a useful marker in evaluation of the functional state of the jaw-neck motor system. PMID- 16708852 TI - Analysis of the interleukin-1 and interleukin-6 polymorphisms in patients with chronic periodontitis. A pilot study. AB - The aim of this study was to analyse whether the interleukin-1 (IL-1) and IL-6 gene polymorphisms were associated with the susceptibility of chronic periodontitis. Genomic DNA was obtained from 20 patients with chronic periodontitis and 31 periodontally healthy subjects. All subjects were of North European heritage. The test subjects were kept in a maintenance program after periodontal treatment but yet showing signs of recurrent disease. Genotyping of the IL-1alpha [+4845C>T], IL-1beta [-3954C>T] and IL-6 [-174G>C] polymorphisms was carried out using an allelic discrimination Assay-by-Design method on ABI PRISM 7900 Sequence Detection System. All genotypes were analyzed using the GeneMapper 2.0 software. A similar distribution of Single Nucleotide Polymorphism (SNP) was seen in both groups. Analysis by logistic regression including gender, IL-1alpha [+4845C>T], IL-1beta [-3954C>T], IL-6 [-174G>C] genotypes, the composite IL-1 genotype, the combination of the composite IL-1 genotype and the IL-6 -174G>C genotype and adjusting for smoking did not result in any statistically significant difference. SNPs in IL-1alpha [+4845C>T], IL-1beta [ 3954C>T] and IL-6 [-174G>C] do not seem to increase the susceptibility to chronic periodontitis in this group of subjects. PMID- 16708854 TI - A 20-year study of dentists' and dental hygienists' assessment of dental caries lesions in bite-wing radiographs. AB - Epidemiological data reveal that the prevalence of dental caries in western countries has decreased in recent decades. The aim of this study was to investigate how dentists and dental hygienists assess dental caries lesions in bite-wing radiographs between 1983 and 2003. All dentists and dental hygienists in Public Dental Health in Uppsala County were offered to take part in the study. The participants assessed manifest and initial caries lesions in eight bite-wing radiographs from three patients individually. An X-ray viewer and binoculars were used. The assessments were repeated in the same radiographs every five years, a total of five times, between 1983 and 2003. In the different test occasions 80 103 dentists and 11-48 dental hygienists participated. The registration of dental caries changed between 1983 and 2003. The number of manifest lesions registered by dentists decreased between 1983 and 1988, but were stable after 1988. Dental hygienists showed no changes in the registration of manifest lesions during the study. Initial lesions registered by dentists and dental hygienists increased between 1988 and 1998. Assessments of initial caries lesions displayed a wider range than manifest lesions. Increasing age and more years in the profession resulted in fewer registered initial caries lesions. Dental hygienists had a tendency to register less caries than dentists. In conclusion, the result of the study indicate that inclusion of initial caries lesions in epidemiological reports should lead to a reduction in reliability. The changes in assessments of manifest caries lesions that took place in the 19805s should be considered when epidemiological data are evaluated. PMID- 16708853 TI - Periodontal conditions in a Swedish city population of adolescents: a cross sectional study. AB - The aim of this epidemiological survey was to analyze the periodontal conditions of 19-year old individuals in an urban area of Sweden, with special reference to gender and socioeconomic factors. A randomized sample of 272 individuals living in Goteborg, Sweden, was clinically examined with regard to oral hygiene, gingivitis, periodontal pockets, probing attachment loss (PAL) and gingival recession. Bitewing radiographs were used for assessments of alveolar bone level (ABL) and dental calculus. A questionnaire-based interview regarding oral hygiene habits was included. Data were analyzed with regard to differences between gender and socioeconomic grouping. The subjects showed a mean plaque score of 59% and a gingivitis score of 44%. 70% of the adolescents had a plaque score of > or = 50%, whereas corresponding figure for gingivitis was 37%. 27% of the subjects had at least one tooth with gingival recession. The mean prevalence of sites with probing depth of > or = 6 mm was 0.5, and the prevalence of PAL > or = 2 mm was 0.7. A radiographic bone level of > or = 2 mm was observed at on average 0.8 teeth per subject. Females had significantly less plaque and gingivitis than males and significantly higher number of teeth with gingival recession. There were no clinically significant differences in periodontal conditions between socioeconomic groups. In conclusion,the survey revealed higher prevalence of plaque and gingivitis among male than female adolescents but no differences between socioeconomic groups. PMID- 16708855 TI - [New era for MS treatment. Severe setback when it comes to enthusiasm--highly effective agent caused serious adverse effects]. PMID- 16708857 TI - [Treatment of chronic pain--a systematic literature review. SBU's summary and conclusions]. PMID- 16708856 TI - [BNP or NT-proBNP should be analyzed in suspected heart failure. Guidelines for analysis and interpretation]. PMID- 16708858 TI - [Stereochemistry and drug effects--a neglected field of knowledge]. PMID- 16708859 TI - [Unclear respiratory symptoms can be pulmonary sequestration]. PMID- 16708860 TI - [Access to health care for asylum seekers in the European Union. A comparative study]. PMID- 16708861 TI - [Neonatal seizure and epileptic attacks in newborn infants. Proposal to a national care program]. PMID- 16708862 TI - [Ambitious measles vaccination program in Vietnam]. PMID- 16708863 TI - [Pain relief in patients treated with Waran]. PMID- 16708864 TI - [WHO survey shows big health status differences between European Union and Eastern Europe]. PMID- 16708865 TI - [Citalopram should not be the first choice]. PMID- 16708866 TI - Evaluation of certain food contaminants. AB - This report represents the conclusions of a Joint FAO/WHO Expert Committee convened to evaluate the safety of various food contaminants, with the aim to advise on risk management options for the purpose of public health protection. The first part of the report contains a general discussion of the principles governing the toxicological evaluation of contaminants and assessments of intake. A summary follows of the Committee's evaluations of technical, toxicological and intake data for certain food contaminants (acrylamide, ethyl carbamate, inorganic tin, polybrominated diphenyl ethers and polycyclic aromatic hydrocarbons). Cadmium was assessed to determine the impact of different maximum limits on intake. Annexed to the report are tables summarizing the Committee's recommendations for intakes and toxicological evaluations of the food contaminants considered, and a description of the statistical methods for dose response modelling as applied at this meeting. PMID- 16708868 TI - Nested ideation and the problem of reality: dreams and works of art in works of art. AB - This paper continues a line of investigation begun in a previous paper on nested dreams and works of art in dreams (Balter 2005). Part I of the present paper seeks to establish that works of art with nested dreams and works of art within them display certain phenomena also observed in comparable dreams: (i) they unsuccessfully deny a painful reality represented in the nested element; (2) they present an antithetical view of that reality (both denying and affirming); and (3) they are consistently associated with the problem of reality (the problem of deciding what is real or true). Part II of this paper seeks to establish the heuristic value of this line of investigation in dreams and art to elucidate the origin of reality testing. PMID- 16708867 TI - Memories of Jacob A. Arlow: A special tribute by the Psychoanalytic Quarterly. PMID- 16708869 TI - The wish for revenge. AB - The wish for revenge is a ubiquitous response to narcissistic injury, and particularly to the narcissistic injury that accompanies oedipal defeat. Vengeful fantasy serves to represent and manage rage and to restore the disrupted sense of self and internalized imagining audience that have resulted from injury. Clinical and literary examples demonstrate the split within the representation of the self and the imagining other that underlies the wish for revenge, and the way that this split operates differently in the psychic economy of the transiently and the chronically vengeful. PMID- 16708870 TI - Unity of analysis: similarities and differences in the analysis of children and grown-ups. AB - When comparing child analysis with that of grown-ups, we are confronted with a substantial unity of psychoanalytic method, beyond the apparent differences. The more we consider the analysis as a here-and-now interaction between analyst and patient with transformative potential, the more age-dependent differences become blurred, and the specificity of a particular analytic couple acquires more significance. At the same time, what seems to be all the same--the actual patient before us--is quite different, with several components (the child, adolescent, and adult parts), implying a need to recognize the part we should address at any given point. PMID- 16708871 TI - What the psychoanalyst learns from his own interpretations. AB - The processes by which the psychoanalyst acquires knowledge of his or her patient exceed the traditional sequence of careful listening and reflection on the meaning of associations. This paper focuses on the value of the analyst's utterances as a source of information about what he or she thinks and is in the process of considering. Movement of the thought process from one subject to another, and the accompanying visual phenomena (among analysts who tend to envision memories and associations), supply valuable data. The author presents several clinical vignettes to illustrate how the analyst discovers ideas and words in the process of giving interpretations. PMID- 16708872 TI - Catching the wrong leopard: courage and masochism in the psychoanalytic situation. AB - This paper introduces the subject of courage into the psychoanalytic discourse about masochism and also demonstrates that ordinary ethical and axiological concerns can and should be included in our psychoanalytic language and practice. At each stage of a psychoanalysis, it may be helpful to consider whether the patient's experience might be that taking a step deeper into the psychoanalytic relationship is both courageous and masochistic. This consideration can open the door to exploration of conscious beliefs and how they are related to unconscious fantasies and assumptions. Considering the possibility that even a sadomasochistic enactment may simultaneously represent a courageous attempt to rework conflict or trauma can enrich the way analysts listen to both manifest and latent material. PMID- 16708873 TI - The misunderstanding (le malentendu). AB - The idea that misunderstanding is simply a problem in both the psychoanalytic situation, between analyst and patient, and in translation between one language and another, is turned on its head in this paper. Originally written in French, this paper addresses both Francophone and Anglophone psychoanalytic communities in the realization that misunderstanding is the ground from which we start in analysis. The author portrays two patients who illustrate that, even when analyst and patient speak the same language, the patient may speak in a style that the analyst must first learn to hear as a kind of dialect, to hear his own misunderstanding, in order for meaning to appear. PMID- 16708874 TI - Reference values in laboratory medicine. PMID- 16708875 TI - Reference values for serum protein and electrolyte study from Rwanda. AB - OBJECTIVE: To estimate reference values of serum proteins and electrolytes in a student population in Butare, Rwanda (altitude: 1768 m; barometric pressure: 629 mm Hg). DESIGN: A laboratory based cross-sectional study. SETTING: The units of physiology and clinical chemistry, department of medical biology, Butare University Hospital and Faculty of Medicine, National University of Rwanda, from February 2002 to May 2003. SUBJECTS: Young healthy adults were selected randomly from the students of the National University of Rwanda, using Epi-Info 6.04 software. RESULTS: The results mean and reference range (2.5th-97.5th percentile) in brackets - are as follows: total proteins: male: 7.3 (6.3-8.4) g/dL, female: 7.3 (6.5-8.5) g/dL; albumin: male: 4.3 (3.1-5.2) g/dL, female: 4.1 (3.2-5.0) g/dL; globulins: male: 3.1 (2.0-4.2) g/dL. female: 3.2 (2.1-4.2) g/dL; sodium: male: 138.5 (130-147) mmol/L, female: 139.8 (132-153) mmol/L; potassium: 4.0 (3.1 5.0) mmol/L both in male and in female; chloride: males: 104.4 (96-112) mmol/L, females: 106.1 (98-114) mmol/L; phosphates: male: 1.13 (0.65-1.59) mmol/L, female: 1.17 (0.71-1.52) mmol/L. CONCLUSION: The values of serum proteins and electrolytes are comparable to classical sea level values; however there is a slight increase in serum chloride. PMID- 16708876 TI - Estimation of the minimum number of Leishmania major amastigotes required for infecting Phlebotomus duboscqi (Diptera: Psychodidae). AB - BACKGROUND: Leishmaniasis is a vector-borne disease in which Leishmania parasites are transmitted by the bite of phlebotomine sand flies. Amastigotes are ingested by the sand fly vector with a blood meal taken from an infected host. This is followed by their differentiation into metacyclic promastigotes which are selectively released and permitted to migrate interiorly so as to make them available for transmission by bite. However, the actual number of amastigotes ingested by the sand fly in the blood meal is not known. OBJECTIVE: To investigate the minimum number of Leishmania major amastigotes required to cause an infection in Phlebotomus duboscqi following an infective blood meal. DESIGN: A laboratory based study. SETTING: Centre for Biotechnology Research and Development, Kenya Medical Research institute, Nairobi. RESULTS: Dissection of all fed sand flies at six days post-infective blood meal revealed that blood containing one amastigote per 0.3 microl in a total volume of 0.5 ml was able to cause an infection in the sand flies, but very few sand flies got infected (7.6% and 9.6% respectively). Concentrations of ten amastigotes per 0.3 microl in 0.5 ml gave infection rates of 35.4% and 26.3% respectively, suggesting that even when the concentration of amastigotes in a bloodmeal was high, not all sand flies feeding on it were able to pick up the parasites. CONCLUSIONS: These observations suggests that one amastigote is sufficient to cause an infection to a sand fly and as a result of multiplication in the gut and the existence of mechanisms that increase the number of infective bites delivered by a female sand fly they are able to sustain the transmission of leishmaniasis in an area. PMID- 16708877 TI - Effects of anti-Leishmania monoclonal antibodies on the development of Leishmania major in Phlebotomus duboscqi (Diptera: Psychodidae). AB - BACKGROUND: Research in our laboratory has previously shown that immune-mediated transmission blocking may be applied to Leishmania infections and that the LPG molecule and anti-LPG monoclonal antibodies was found to be an excellent candidate against L. major infections. OBJECTIVE: To test the effect of monoclonal antibodies (MABs) raised against different species of Leishmania for their ability to inhibit development of Leishmania major in Phlebotomus duboscqi sand flies. DESIGN: A laboratory based study. SETTING: Centre for Biotechnology Research and Development, Kenya Medical Research Institute, Nairobi. RESULTS: Sand fly dissections on days two, four and six post-feeding showed that monoclonal antibodies against L. donovani (Ld2cb and Ld3A3) were the most effective at inhibiting L. major development than those raised against L. aethiopica, L. major or L. tropica. Ld2cb inhibited L. major development by 82% in sand flies fed on 1 x 10(6) amastigotes while Ld3A3 inhibited by 72%; 58% and 74% in those fed on 1 x 10(5) amastigotes respectively. Monoclonal antibodies against L. aethiopica (Lae 3c6) inhibited L. major development by 28% and 40% for sand flies fed on 1 x 10(6) and 1 x 10(5) amastigotes respectively. Anti-L. major monoclonal antibody (Lm5A5) inhibited L. major development by 16% in sand flies fed on 1 x 10(6) amastigotes and 25% in sand flies fed on 1 x 10(5) amastigotes. Anti-L. tropica antibody (Lt2c8) inhibited L. major development in P. duboscqi fed on 1 x 10(6) by 28 %and 33% in those fed on 1 x 10(5) amastigotes. Most of the parasites seen in sand flies which fed on L. donovani mABs (Ld2cb and Ld3A3) were nectomonads and a few haptomonads. In all the control groups, parasite development followed the normal developmental stages up to the metacyclic stage. In sand fly groups fed on monoclonal antibodies raised against L. aethiopica, L. major or L. tropica there was limited parasite development inhibition, and the promastigotes developed and migrated forward in a normal pattern as observed in the controls. CONCLUSIONS: These results suggests a possible role of humoral mechanisms in protection against leishmaniasis and potentially useful in reducing parasite development in the sand fly. PMID- 16708878 TI - Iatrogenic ureteric and bladder injuries in obstetric and gynaecologic surgeries. AB - OBJECTIVE: To review iatrogenic ureteric and urinary bladder injuries from obstetric and gynaecological surgeries treated in the urology department analysing; ureteric anatomy, aetiologic factors, diagnosis, treatment and outcomes. DESIGN: A retrospective study. SETTING: Institute of Urology, Kilimanjaro Christian Medical Center (KCMC), Moshi, Tanzania. SUBJECTS: Twenty three women were treated for iatrogenic ureteric and bladder injuries secondary to obstetric and gynaecological procedures in the department of urology between June 1994 and July 2004. RESULTS: Hysterectomy was the leading cause of ureteric injuries contributing to nine (47.4%) of the 19 ureteric injuries. Caesarian sections were the second leading cause and contributed 6(31.6%) of 19 ureteric injuries. Vesical vaginal fistula (VVF) repairs lead to two (10.5%) ureteric injuries one of which was bilateral. There was a case each (5.3%) from ovarian cystectomy and forceps delivery. There were only three cases of intra-operative diagnosis of ureteric injuries. There were four bladder injuries half of which came from hysterectomy. The leading definitive urological treatment for ureteric injury was ureteric re implantation and all kidneys were saved. CONCLUSION: Iatrogenic ureteric and bladder injuries from gynaecologic surgeries are globally rare but are liable to occur due to the inherent ureteric anatomic factors in the pelvis. Intra-operative diagnosis of injury is a rare feature. The practical principles to prevent and repair ureteric injuries have been presented and discussed. The true risk to the patient lies in delayed, missed diagnosis and inadequate treatment. Endourologic techniques offer an alternative diagnostic and treatment method for women previously injured in open pelvic surgeries. PMID- 16708879 TI - Hepatitis A seroepidemiology in Eastern Turkey. AB - BACKGROUND: It has been reported that there are great deal of changes in the epidemiology of hepatitis A at the present. The epidemiologic studies regarding to hepatitis A virus (HAV) infection are not sufficient both in the country and study region. OBJECTIVE: To investigate HAV seroprevalence and association with socio-demographic variables. DESIGN: Cross-sectional and community based study. SETTING: Erzurum Province, Eastern Turkey between April and June 1998. SUBJECTS: Four hundred and fifty persons under the age of 30 years. RESULTS: The rate of the study involvement was 87.1% (392 persons) and ratio for seropositivity 84.2%. Although seropositivity is elevated by age, it has been high within the younger age group. Epidemiological changes could not be interpreted, since the study results were first for the group of people aged under 30 years. Statistical significance was found between hepatitis A seroprevalence and age, the number of households and localisation of toilet within the house. There was no correlation between hepatitis A and sex, jaundice history, family type, tap water availability in the residential area. CONCLUSION: The frequency of hepatitis A infection appearance in the region was found out to be high as compared to other regions of Turkey. PMID- 16708880 TI - Seroprevalence of hepatitis B and C viruses among women of childbearing age in Moshi Urban, Tanzania. AB - OBJECTIVES: To describe the seroprevalence of hepatitis C and B viruses and their association with HIV and other sexually transmitted diseases (STDs) among women aged 15-49 years, attending primary health care clinics in Moshi urban, Tanzania. DESIGN: A cross-sectional study. SETTING: Three primary health care clinics in Moshi, Tanzania. SUBJECTS: A total of 382 consenting women attending reproductive and child health clinics between September-December 1999. RESULTS: The seroprevalence of anti-HCV was 1.0%, for HBsAg 4.2% and for HIV 11.5%. HIV infection and other sexually transmitted diseases were not associated with anti HCV or HBsAg. There was no interrelation between HCV and HBV markers. CONCLUSION: Hepatitis C Virus infection is infrequent among women in urban Tanzania. HCV and HBsAg were not more prevalent in HIV-infected women. Public preventive efforts should thus focus on hepatitis B virus immunisation. PMID- 16708881 TI - Congenital malformations in the North-Eastern Democratic Republic of Congo during Civil War. AB - OBJECTIVE: To report on changes in the incidence of congenital malformations in a context of an ongoing civil conflict. DESIGN: Retrospective study. SETTING: Centre Medical Evangelique (CME) of Nyankunde, a 250 bed referral hospital, in the North Eastern Democratic Republic of Congo (DRC) during the period 1993 to 2001. SUBJECTS: A total of 8824 babies were delivered alive at our maternity; and 36 of them (0.41%) were born with a clinically diagnosed congenital malformations. RESULTS: The breakdown of the observed malformations was as follow: clubbed foot nine, congenital hydrocephalus eight, spina bifida six, cleft lip four, encephalocele two, syndactyly two, imperforated anus two, Anencephaly one, lymphangioma one, bladder exstrophy one. There was a significant increase in the annual incidence of congenital malformations (p<0.001). CONCLUSION: Ongoing civil conflicts or wars ultimately have a negative impact on the incidence of congenital malformations. This is an indirect and multifactorial consequence. We conclude that ongoing civil conflict is a contributing factor to the increasing incidence of congenital malformations seen in the North-Eastern DRC. PMID- 16708882 TI - Oral health status among Kenyans in a rural arid setting: dental caries experience and knowledge on its causes. AB - OBJECTIVE: To determine the dental caries experience and knowledge on the causes and preventive measures for dental diseases. DESIGN: A community based cross sectional descriptive study. SETTING: Elwak sub-district hospital, North Eastern Province, Kenya. SUBJECTS: One hundred and forty one adults who presented themselves during a dental check up at a sub-district hospital and gave written consent. MAIN OUTCOME MEASURES: Dental caries status and knowledge on its causes and preventive measures. The importance of outreach programmes in obtaining information as well as helping to alleviate the pain and suffering caused by dental diseases among communities living in remote areas is also revealed. RESULTS: Of the one hundred and forty one individuals, who were included in the study, 63.1% were women and 36.9% were men. Their ages ranged between 18 and over 65 years. 41.1% were in the 18-24-year age bracket. Regarding the oral health knowledge, 43% did not know any causes of dental diseases while 36%, 17% and 12% knew that diet, "dirt" on teeth and bacteria were possible causes, respectively. Fifty percent did not know any preventive measures for dental diseases while the rest indicated abstention from the consumption of sugary foods; and only 0.8% mentioned use of fluoridated toothpaste as a preventive measure for dental caries. 56.7% of the subjects were caries free. The mean DMFT for all ages was 3.4. Of those with caries 72.1% were women. CONCLUSION: There is a low level of oral health awareness and a moderately high level of dental caries experience in this community with women apparently carrying the biggest burden of dental caries. PMID- 16708883 TI - Evidence for expression of endogenous retroviral sequences on primate reproductive tissues and detection of cross-reactive ERVS antigens in the baboon ovary: a review. AB - OBJECTIVE: To review recent research findings on the specific expression of endogenous retroviral sequences (ERVS) in reproductive tissues and their possible physiological roles. ERVS have been implicated in several biological events such as induction of resistance to exogenous retrovirus invasion, involvement in placental trophoblast formation, sperm maturation and differentiation; and stimulation of local immunosuppression to protect the foetus from immunological attack. DATA SOURCES: Critical review of relevant articles and abstracts cited in international and local journals, literature searches on Medline and Medchem up to 2005. DATA SYNTHESIS: Retroviruses have been implicated in the induction of tumour and immunological disorders. Over the years, endogenous retroviruses (ERVs) and retroviral elements have been detected in the genome of many vertebrate species, including primates. The evidence for the presence of retroviruses in the primate tissues such as the placenta, ovary, breast, testis and epididymis has been documented using electron microscopic studies. Retrovirus like particles were found budding from the basal membrane of syncytiotrophoblasts, as well as in tumour cell lines in embryonic carcinoma or teratocarcinomas. Apart from their pathological effects, recent evidence suggests that these ERVs may play useful roles in normal physiological events. RESULTS: Recent studies indicate the expression of endogenous retroviruses in the testis, epididymis, placenta and breast. However, limited data exist on the detection of ERVs in the ovary. Overall, the precise functions for ERVs in these tissues are not well understood. In the testis and epididymis, speculative functions may include among others spermatogenesis and/or sperm maturation (differentiation) whereas in placenta they are possibly associated with trophoblast fusion and locally induced immunosuppression to protect the foetus from immunological attack. Experiments in our laboratory have indicated restricted expression of retroviral antigens including baboon endogenous retroviral proteins (BERV), ERV 3, HIV-1 gp41 and HERV-K env in the baboon ovary. CONCLUSION: ERVs are specifically expressed in different mammalian reproductive tissues and may have unique physiological roles. PMID- 16708884 TI - Health professionals' readiness for infant hearing screening in Lagos, Nigeria. AB - The authors set-out to ascertain the readiness of professional healthcare workers in Nigeria for the implementation of the country's new national health policy, which for the first time, seeks to address the impact of infant hearing loss on early childhood development through early detection and timely intervention. A cross-sectional survey of nurses and doctors showed that health professionals had positive attitudes towards early hearing detection but were limited in their knowledge about current possibilities with infant screening. The need for relevant/on-going professional education is indicated to effectively promote hearing screening services in early childhood. PMID- 16708885 TI - Re: Acute colonic pseudo obstruction (Ogilvie's syndrome): a case report. PMID- 16708886 TI - Lipid peroxidation in systemic lupus erythematosus. AB - Lipid peroxidation is an important process in oxygen toxicity. Free radicals inflict this damage by attacking polyunsaturated fatty acids, thus setting off a deleterious chain reaction that ultimately results in their disintegration into malondialdehye, 4 hydroxy-2-nonenal and other harmful by-products. Peroxidation of lipids has been implicated in several diseases including systemic lupus erythematosus (SLE). SLE is an autoimmune disorder with unknown aetiology, characterized by the presence of autoantibodies to self-antigens. There is a significant increase in the production of free radicals like superoxide and hydroxyl radicals in SLE. Indices of lipid peroxidation, like conjugated dienes, malondialdehyde, 8-isoprostaglandin F2 alpha are significantly elevated in SLE. Increased ceruloplasmin levels and decreased transferrin levels in the sera of SLE patients have also been described. The activities of the antioxidant enzymes superoxide dismutase, catalase and glutathione peroxidase and the amounts of the antioxidant reduced glutathione are also significantly altered in this disease. In addition, there are significant changes in the essential fatty acid profile in the sera of those affected with the disease. In animal models of the disease, immunization of mice with peptides derived from autoantigens induces SLE like disease. Immunization with an oxidatively modified autoantigen led to the rapid development of autoimmunity compared to immunization with the unmodified autoantigen. Thus, oxidative damage appears to play an important role in SLE pathogenesis. PMID- 16708888 TI - Secretory factors of human neuroblastoma (IMR-32) and human glioblastoma (U87MG) cell lines induce neurite outgrowths in PC12 cells. AB - Neurite outgrowth is essential for the communication of the nervous system. The rat Pheochromocytoma (PC12) cells are commonly used in the neuronal cell study. It is well known that exogenous stimuli such as Nerve Growth Factor (NGF) induce neurite outgrowth. In the present study it has been investigated whether or not the conditioned medium from human neuroblastoma cell line (IMR-32) and human glioblastoma cell line (U87MG) may augment neurite outgrowth in PC12 cells. PC12 were cultured with and without conditioned media of IMR-32 and U87MG. The result showed that both the conditioned media induce neurite outgrowth within 48 hr and stops further proliferation of PC12 cells. However no outgrowth was noted in PC12 cells incubated without conditioned medium. In conclusion, it is shown that both the conditioned media (IMR-32 and U87MG) have the potential to induce the neurite outgrowth in the PC12 cells. PMID- 16708887 TI - Nanotechnology based drug delivery system(s) for the management of tuberculosis. AB - The era of nanotechnology has allowed new research strategies to flourish in the field of drug delivery. Nanoparticle-based drug delivery systems are suitable for targeting chronic intracellular infections such as tuberculosis. Polymeric nanoparticles employing poly lactide-co-glycolide have shown promise as far as intermittent chemotherapy in experimental tuberculosis is concerned. It has distinct advantages over the more traditional drug carriers, i.e. liposomes and microparticles. Although the experience with natural carriers, e.g. solid lipid nanoparticles and alginate nanoparticles is in its infancy, future research may rely heavily on these carrier systems. Given the options for oral as well as parenteral therapy, the very nature of the disease and its complex treatment urges one to emphasize on the oral route for controlled drug delivery. Pending the discovery of more potent antitubercular drugs, nanotechnology-based intermittent chemotherapy provides a novel and sound platform for an onslaught against tuberculosis. PMID- 16708889 TI - Beta-adrenoceptor agonist treatment reverses denervation atrophy with augmentation of collagen proliferation in denervated mice gastrocnemius muscle. AB - Daily oral administration of isoproterenol hydrochloride (60 mg/kg body weight; for 30 days) a beta-receptor agonist to normal innervated and denervated adult male Swiss albino mice confirmed its ability to induce skeletal muscle hypertrophy and reverse denervation atrophy respectively. Measurement of total tissue proteins and dry muscle mass showed 15-17% increase with 6% rise of hypertrophy index in gastrocnemius muscle. Hydroxyproline assay employed to measure the total tissue collagen exhibited 45% increase in collagen in normal innervated gastrocnemius muscle in response to beta agonist treatment. beta adrenoceptor agonist ameliorated denervation atrophy along with further increase in collagen content of denervated gastrocnemius muscle. PMID- 16708890 TI - Hydrogen peroxide commences copper induced DNA damage isolated from human blood: in vitro study. AB - The present study revealed the damaging effects of copper and hydrogen peroxide on DNA isolated from human blood in in vitro. Ultra violet spectral studies showed that copper and H2O2 alone (at 20 mM) caused destabilization of DNA structure. Notwithstanding, the effect was more prominent in combination of copper and H2O2. Further, agarose gel electrophoretic studies revealed that neither copper nor H2O2 alone had DNA fragmentation (up to 40 mM concentration), while copper and H2O2 together caused massive DNA fragmentation even at lower concentrations (4 mM copper + 4 mM H2O2). Therefore, it was concluded from the present study that the observed destabilization of DNA associated with alterations in configuration and subsequently massive DNA fragmentation was in response to copper and H2O2. Further, fluorescence spectroscopy and TUNNEL assay will address destabilization and fragmentation of naked DNA more precisely. PMID- 16708891 TI - Differential acetylcholinesterase activity in rat cerebrum, cerebellum and hypothalamus. AB - Acetylcholinesterase (AChE) has been purified from three different regions of rat brain using Sephadex G 200 column. SDS PAGE (6%) showed single band for the purified AChE fractions. Purified and lyophilized AChE from different (NH4)2SO4 precipitated fractions of three brain parts were utilized for in vitro enzyme kinetics using Dimethoate (Dmt) as inhibitor. K(m) values for cerebellum and hypothalamus were almost similar whereas cerebrum showed a different K(m) value compared to other two regions. With the drug Rivastigmine it was found that % G1 and G4 forms of AChE in three different parts of brain are different. PMID- 16708892 TI - Comparative evaluation of possible ocular photochemical toxicity of fluoroquinolones meant for ocular use in experimental models. AB - Fluoroquinolones (FQs) are extensively used in bacterial keratitis and other intraocular infections. Since eye is constantly exposed to light, incidence of ocular phototoxicity due to commonly used FQs is of great interest for their safe use. Phototoxicity of commonly used FQs (ciprofloxacin, lomefloxacin, pefloxacin, ofloxacin, sparfloxacin and gatifloxacin), has been evaluated by using HET-CAM-UV model (Photo Hen Egg Test-C Chorioallantoic Membrane model). This study was further extended by adding lomefloxacin dissolved in bovine vitreous (0.5 ml) on the chorioallantoic membrane (CAM). Using a standard scale, the phototoxic damage was assessed at different time intervals. Respective controls were kept in dark to distinguish the toxicity of the drugs per se. The results showed that the phototoxicity induced by lomefloxacin was very high followed by gatifloxacin and sparfloxacin and least for other drugs studied. Interestingly, lomefloxacin along with vitreous showed significantly low phototoxicity. This could be due to the antioxidant property of ascorbic acid present in the vitreous. PMID- 16708893 TI - Development of biosensor based on immobilized L-glutamate oxidase for determination of monosodium glutamate in food. AB - A monosodium glutamate (MSG) biosensor with immobilized L-glutamate oxidase (L GLOD) has been developed and studied for analysis of MSG in sauces, soup etc. The immobilized enzymatic membrane was attached with oxygen electrode with a push cap system. The detection limit of the sensor was 1 mg/dl and the standard curve was found to be linear upto 20 mg/dl. Response time of the sensor was 2 min. Cross linking with glutaraldehyde in presence of Bovine Serum Albumin (BSA) as a spacer molecule has been used for immobilization. Optimization of the sensor was done with an increase in L-GLOD concentration (6.3-31.5 IU) and also with increase in loading volume of enzyme solution (5-20 microl). Optimization of pH and temperature was also studied. The permeability of O2 through different membrane was studied with and without immobilized L-GLOD. The enzymatic membrane was used for over 20 measurements and stability of the membrane was observed. PMID- 16708894 TI - Hepatoprotective activity of Hemidesmus indicus R. br. in rats. AB - Treatment of rats with paracetamol and CCl4 produced a significant increase in the levels of serum glutamate pyruvate transaminase (SGPT), serum glutamate oxaloacetate transaminase (SGOT), alkaline phosphatase (ALP), total and direct bilirubin. Rats pretreated with methanolic extract of roots of H. indicus (100 500 mg/kg body weight, po) exhibited rise in the levels of these enzymes but it was significantly less as compared to those treated with paracetamol or CCl4 alone. The results of methanolic extract of H. indicus were comparable with the standard hepatoprotective agent silymarin (100 mg/kg). Maximum hepatoprotective effect was found to be at the dose of 250 mg/kg body weight in case of CCl4 induced hepatic damage while 500 mg/kg body weight in case of paracetamol induced hepatic damage. The results suggest that methanolic extract of H. indicus roots possesses a potential antihepatotoxic activity. PMID- 16708895 TI - Antioxidant activity of Arthritin--a polyherbal formulation. AB - In complete freund's adjuvant induced arthritis in male albino rats, a significant increase in serum lipid peroxidase besides increase in paw swelling and a significant decrease in superoxide dismutase, glutathione peroxidase and total reduced glutathione levels were observed. Arthritin produced a marked reversal of these enzyme levels, besides a significant reduction in paw swelling. The results suggest that, the polyherbal formulation 'Arthritin' exerts its effects by modulating lipid peroxidation and enhancing anti-oxidant and detoxifying enzyme systems. PMID- 16708896 TI - Photosynthetic acclimation to elevated CO2 in relation to Rubisco gene expression in three C3 species. AB - Wheat (Triticum aestivum L. var. DL 1266-5), sunflower (Helianthus annuus L. var. MSFH 17) and mungbean [Vigna radiata (L.) Wilczek var. P 9072] were grown in field under atmospheric (360 +/- 10 cm3 m(-3), AC) and elevated (650 +/- 50 cm3 m(-3), EC) CO2 concentrations in open top chambers for entire period of growth and development. Photosynthetic acclimation to elevated CO2 was examined by comparing photosynthesis rate (Pn), Pn/Ci curves, leaf contents of RuBP carboxylase/oxygenase (Rubisco), change in the transcripts of Rubisco small subunit (SSU) gene and leaf carbohydrate constituents in AC and EC grown plants. The study indicated that photosynthetic acclimation to elevated CO2 concentration in wheat occurred because of down regulation of Rubisco, through limitation imposed on Rubisco SSU gene expression, as a consequence of sugar accumulation in the leaves. Leaf starch accumulators, sunflower and mungbean, showed no down regulation of Pn under EC. The Rubisco contents (%) in leaf soluble protein and rbcS transcript levels were not significantly affected in EC plants compared to AC plants of sunflower and mungbean. The study indicated that accumulation of excess assimilates in the leaves as starch was less inhibitory to Pn and would, therefore, be an important trait for sustenance of Pn not only under EC, but also under AC, where Pn inhibited by end products. PMID- 16708897 TI - Modification of biochemical parameters of gentamicin nephrotoxicity by coenzyme Q10 and green tea in rats. AB - The present study was designed to investigate the possible potential protective role of coenzymeQ10 (CoQ10; 10 mg/kg/day, ip) and/or green tea (GT; 25mg/kg/day, po) against gentamicin (GM) nephrotoxicity. Marked increase in the level of serum urea. creatinine and lipid peroxidation (LPO) content was found after administration of gentamicin (80 mg/kg/day, ip) for eight days along with significant decrease in the antioxidant enzymes, superoxide dismutase (SOD), reduced glutathione (GSH), catalase (CAT) as well as brush border enzymes (Na+/K+ ATPase, Mg(+2)ATPase and Ca2+ ATPase). Treatment with CoQ10 or green tea alone with GM showed significant decrease in serum urea, creatinine and tissue LPO content and significant increase in antioxidant and membrane bound enzymes. Combined treatment with CoQ10 and green tea was more effective in mitigating adverse effect of GM nephrotoxicity. The present work indicated that CoQ10 and green tea due to their antioxidant activity modified the biochemical changes occurred during gentamicin nephrotoxicity and thus had a potential protective effect. PMID- 16708898 TI - Cross infectivity of baculovirus, Spilarctia obliqua nuclear polyhedrosis virus against mulberry pest, Porthesia xanthorrhoea Kollar. AB - Spilarctia obliqua nucleopolyhedrosis virus (SoNPV) has been found effective against S. obliqua (Walker) as well as Porthesia xanthorrhoea Kollar. LC50 concentration of S. obliqua SoNPV against S. obliqua and P. xanthorrhoea was 2.5 x 10(4) and 3.7 x 10(4) POBs/ml, respectively. LT50 of SoNPV against S. obliqua was 5.73 days, while it was 6.98 days for P. xanthorrhoea. Cross infectivity of SoNPV against P. xanthorrhoea and ease of its mass production may make it an ideal biopesticide. PMID- 16708899 TI - Analgesic activity of leaf extracts of Culcasia scandens P. Beauv. AB - Analgesic activity of methanol leaf extract of C. scandens obtained by column chromatography and its graded solvent fractions, was evaluated in mice using acetic acid-induced abdominal writhing and formalin-induced paw licking. The extract and fractions significantly inhibited abdominal writhing and two phases of formalin-induced paw licking in mice, indicating that antinociceptive activity may involve inhibition of pain by peripheral and central mechanisms. PMID- 16708900 TI - Individual account retirement plans: an analysis of the 2004 survey of consumer finances. PMID- 16708901 TI - Classical fever of unknown origin (FUO): current causes in Mexico. AB - BACKGROUND: Causes of FUO change according to medical innovations, modifications of social circumstances, and emerging health risks. Aim. To describe the epidemiology of classical FUO, the time and procedures to achieve a definitive diagnosis, and to underline the variables useful in distinguishing FUO categories. Setting. A third-referral center in Mexico City. METHODS: Patients admitted with prolonged fever were evaluated. Clinical charts of patients with classical FUO were assessed; comparisons between classical FUO categories were made. RESULTS: 45 patients with 44.9 +/- 17.2 years of age, previous fever duration of 51.2 +/- 51.5 days, and 88.9% referred from other hospitals were evaluated. Nineteen patients had infectious causes; eight, neoplastic conditions; 12, inflammatory non-infectious diseases; one had another cause, and five were discharged with no etiologic diagnosis. Age, LDH levels, length of fever, and weight loss greater than 10 kg may be used to classify patients into a definite category. CONCLUSIONS: Classical FUO is an unusual presentation of frequent infectious diseases; SLE is the main cause within the inflammatory non-infectious conditions, and non-Hodgkin's lymphoma is the first cause of cancer. Some clinical and laboratory clues may be used to guide the study work up of patients with classical FUO. PMID- 16708902 TI - Incidence and risk factors for cutaneous adverse drug reactions in an intensive care unit. AB - OBJECTIVE: To evaluate the incidence of adverse cutaneous drug reactions in intensive care unit patients. DESIGN: Cohort study. SETTING: General adult intensive care unit of an institutional tertiary care hospital. PATIENTS: Patients in the intensive care unit during a consecutive 8-month period were examined for adverse cutaneous drug reactions. RESULTS: Patients in the intensive care unit have an incidence of 11.6% of adverse cutaneous drug reactions. Associated risk factors were female gender, obesity, age over 60 and immune dysregulation (systemic lupus erythematosus, dysthyroidism, and antiphospholipid antibodies syndrome). Few patients had previous history of adverse cutaneous drug reactions. Antimicrobials were the main drug involved. Morbilliform rash followed by urticary were the most frequently observed reactions. Interestingly, over 50% of patients with massive edema -independent of etiology- died. CONCLUSIONS: Intensive care unit patients are particularly at risk for developing an adverse cutaneous drug reaction. PMID- 16708903 TI - [Critical reading skill goes along with the proximity to health research?]. AB - OBJECTIVE: The evaluation of a reading skill to critically read theoretical texts, interpretation; at different groups linked to investigation and, explore if a closer, longer experience with this, can influence this skill development. MATERIAL AND METHODS: An instrument that comprises a text on epistemologic aspects and the relation of science with society, was validated. It was applied on four natural groups, selected by a degree of involvement they had shown in health research: students at "pre-degree" (n = 23); students at post-degree (n = 19), academic administrative assessors (n = 17), and professional researchers (n = 20). RESULTS: Reliability coeficients of the instrument were 0.91 y 0.94. From 74 as maximum score expected, medians of groups were 19 for pre-grade and post grade students, 17 for assessors, and 21 for researchers (p = 0.50). CONCLUSIONS: The not sufficient ability to critically read a text, that we supposed was more or lest related to all studied groups, come to be highly relevant, when a need for the researcher to acquire plain understanding, of the importance of his actions is acknowledged. And on the other hand, the fact that students on upper levels, be conscious of the reason why science is beforehand included in their education. PMID- 16708904 TI - [Developing undergraduate medical boarding's clinical aptitude in anemic deficiencies with a participation promotive educational strategy]. AB - OBJECTIVE: To compare the profit of a participation promotive educational strategy versus traditional teaching on development of the undergraduate medical boarding's clinical aptitude. METHODS: A quasiexperimental design was made. An instrument was built and validated in order to explore clinical aptitude. It contains three actual clinical cases about iron, folic acid or cobalamin deficiency, and a questionnaire with 164 items to explore eight clinical aptitude indicators; experts attending and teaching physicians validated it. A Kuder Richardson coefficient of 0.80 was gotten when we estimated the internal reliability. The instrument was applied simultaneously to 55 undergraduate medical boarding from two hospitals of the Instituto Mexicano del Seguro Social the starting year of undergraduate boarding (initial measuring) and at finish once again (end measuring). The experimental group (GE, n = 29) was exposed to a participation promotive educational strategy, and control group (GC, n = 26) to a traditional one. RESULTS: The overall scores of initial measuring in the GE and GC were low: 57 y 42 respectively (maximum: 164); at end measuring an increase in overall scores was observed: in the GE 68 (p < 0.02) and GC 47 (NS). CONCLUSIONS: The participation promotive educational strategy is better than traditional teaching for developing the undergraduate medical boarding's clinical aptitude. It must be considerated to propitiate changes to develop a more reflexive and critical education. PMID- 16708905 TI - [Ultra structure of retinopathy induced by hyperoxia in developing rats]. AB - In order to evaluate the effect of postnatal hyperoxia on retinal structure, newborn rats were exposed to different oxygenation intervals (80 +/- 1%) with three interruptions of 21% (30 min each). Four groups of rats were exposed from birth to the 6th, 9th, 12th and 14th postnatal day, respectively and another group was placed under normoxia. After this period all oxygenated groups and the controls remained under normoxia until they were 30 days old for the structural analysis of retina. Retinal histology was carried out using conventional techniques for transmission electron microscopy (TEM). In the ganglion cell layer of the retina from rats exposed for 9 days to hyperoxia, capillaries with large projections toward the lumen, were observed as a possible consequence of cellular edema of endothelium. The most severe damage was observed in rats exposed to hyperoxia during 12 and 14 days, showing mitochondrias swollen up and without crests in the areas surrounding the capillaries, necrosis and apoptosis processes, dense bodies, cells with swollen cytoplasms and rupture of the plasmatic membrane. The results suggest that postnatal hyperoxia causes severe damages to the retina in developing rats with a direct relationship between the time exposed to oxygen and ultra structural damages. PMID- 16708906 TI - [High molecular weight kininogen in inflammation and angiogenesis: a review of its properties and therapeutic applications]. AB - The plasma kallikrein-kinin system (KKS) participates in the pathogenesis of inflammatory reactions involved in cellular injury, coagulation, fibrinolysis, kinin formation, complement activation, cytokine secretion and release of proteases. It has been shown that KKS activation in the systemic inflammatory response syndrome results in decrease of its component plasma proteins. Similar changes have been documented in diabetes, sepsis, children with vasculitis, allograft rejection, disseminated intravascular coagulation, patients with recurrent pregnancy losses, hereditary angioedema, adult respiratory distress syndrome and coronary artery disease. Direct involvement of the KKS in the pathogenesis of experimental acute arthritis and acute and chronic enterocolitis has been documented by previous studies from our laboratory using experimental animal models. It has been found that in HK deficient Lewis rats, experimental IBD was much less severe. We showed a genetic difference in kininogen structure between resistant Buffalo and susceptible Lewis rats, which results in accelerated cleavage of HK and it is responsible for the susceptibility to the inflammatory process in the Lewis rats. It has been demostrated that therapy with a specific plasma kallikrein inhibitor (P8720) modulated the experimental enterocolitis, arthritis and systemic inflammation. Furthermore, it has been shown that a bradykinin 2 receptor (B2R) antagonist attenuates the inflammatory changes in the same animal model. We have showed that a monoclonal antibody targeting HK decreases angiogenesis and arrests tumor growth in a syngeneic animal model. In summary, these results indicate that the plasma KKS plays a central role in the pathogenesis of chronic intestinal inflammation, arthritis and angiogenesis. PMID- 16708907 TI - [The carcinoembryonic antigen: apropos of an old friend]. AB - The carcinoembryonic antigen (CEA) is glycoprotein localized in the apical surface of mature enterocytes. The members of the CEA gene family are clustered on chromosome 19q13.2. It is formed by 29 genes, of which 18 are expressed. Many functions of CEA have been known in healthy indiuiduals, however its role as cell adhesion molecule is the most studied. Besides the colon, CEA is expressed in the stomach, tongue, oesophagus, cervix, and prostate. The most important clinical function is in colorectal, gastric and ovary cancer. It is used as prognosis marker, staging system, recurrence, treatment response and liver metastases. There are many non-neoplasic diseases that enhance CEA value. Actually, CEA is being studying as target of immunotherapy. PMID- 16708908 TI - [Basal homocysteine blood levels in patients with venous thromboembolic disease from a rural area of southern Extremadura, Spain]. PMID- 16708909 TI - [Comments to the manuscript: Does cranberry juice have bacteriostatic activity?]. PMID- 16708910 TI - [Treatment strategies for chronic myeloid leukemia]. PMID- 16708911 TI - [Childhood acute lymphoblastic leukemia with t(1;19) lacking E2A-pBX1 chimeric transcripts]. AB - We present a pediatric case of acute lymphoblastic leukemia (ALL) with chromosomal translocation 1;19 lacking E2A-PBX1 chimeric transcripts. On admission, the patient showed remarkable splenomegaly. Laboratory findings demonstrated that WBC was 12900/microl with blasts 61.5%. Bone marrow examination revealed 1282 X 10(3)/microl of the nucleated cell count with 95.5% lymphoblasts. Surface marker analysis showed an early pre-B lineage immunophenotype (CD10+, CD19+, CD34+, surface Ig-). Although G-banding chromosomal analysis showed 46,XY,der(19)t(1;19)(q23;p13), E2A-PBX1 chimeric transcripts and E2A gene rearrangement were not detected with the polymerase chain reaction method and Southern blot analysis, respectively. The patient was assigned to high-risk ALL according to the criteria of the Japan Association of Childhood Leukemia Study. His clinical response to prednisolone monotherapy for the initial 7 days and subsequent multidrug chemotherapy was excellent, and he achieved complete remission on day 15, which has lasted for more than 30 months. We reviewed the bibliography of the clinical and biological features of 17 children with t(1;19)+E2A-PBX1- ALL including this case. The two prominent characteristics included an early pre-B immunophenotype (11/13) and hyperdiploid (>50 chromosomes) chromosome abnormality (8/14). However, there was substantial heterogeneity in the demographic features and prognosis. Further accumulation of such patients will facilitate the determination of the appropriate treatment for childhood t(1;19)+E2A-PBX1- ALL. PMID- 16708912 TI - [Mantle cell lymphoma mimicking chronic lymphocytic leukemia]. AB - A 62-year-old male was admitted to our hospital complaining of dyspnea in March, 2002. He had remarkable bone marrow invasion with a significant number of leukemic cells, anemia and thrombocytopenia. In addition he had generalized lymphadenopathy including a bulky mass in the left cervix. Surface marker analysis of abnormal cells showed CD 5+, 10-, 19+, 20+, 23+, and kappa+, and immunohistochemistry revealed cyclin D1-positive cells. Chromosome analysis showed del(11q). The patient was diagnosed as having mantle cell lymphoma, stage IVB, and received combination chemotherapy. He could not obtain complete remission and died after 29 months. We found it very difficult in this case to make a differential diagnosis between mantle cell lymphoma and chronic lymphocytic leukemia. We report on this case and summarize the problem of the differential diagnosis. PMID- 16708913 TI - [Clinical significance of anti-phosphatidylserine-prothrombin complex antibodies in antiphospholipid syndrome]. AB - We clarified the clinical significance of IgG anti-phosphatidylserine-prothrombin complex (PS-PT) antibodies in the antiphospholipid syndrome (APS). The study population consisted of 122 patients with SLE and lupus-like disease. IgG anti-PS PT antibodies were detected in 44% of 59 patients according to the diagnostic criteria by Harris and Hughes. This frequency was significantly (p < 0.005) higher than the 14% seen in patients without APS. IgG anti-PS-PT antibodies were strongly (p < 0.005) associated with thrombosis. In addition, IgG anti-PS-PT antibodies were positive in 64% of IgG beta2-GPI dependent anti-cardiolipin antibody negative APS patients under the Sapporo criteria. The above findings indicate that IgG anti-PS-PT antibodies as well as beta2,-GPI dependent anti cardiolipin antibodies should be examined in the diagnosis of APS. PMID- 16708914 TI - Poincare's criterion in room acoustics. PMID- 16708917 TI - Fast calculation system specialized for head-related transfer function based on boundary element method. AB - Recently, development of a numerical calculation of the head-related transfer function (HRTF) has been conducted using a computer model of a human head and the boundary element method. The reciprocity theorem is incorporated into the computational process in order to shorten the computational time, which is otherwise very long. On the other hand, another fast HRTF calculation method for any source position, which is realized by calculating factors independent of the source position in advance, has been suggested by the authors. Using this algorithm, the HRTF for any source position can be obtained in a few seconds with a common PC. The resulting HRTFs are more precise and are calculated faster than those by using the reciprocity theorem. However, speeding the process up even further is required in order to respond to a head movement and rotation or to moving sources during binaural sound reproduction. In this paper, a faster calculation method by incorporating a time domain operation into the authors' previous algorithm is proposed. Additionally, the new formulation, which eliminates the extra computational time in the preprocess, is proposed. This method is shown to be faster than the previous ones, but there are some discrepancies at higher frequencies. PMID- 16708919 TI - Paranasal sinus ventilation by humming. AB - The gas nitric oxide (NO) is generated in the human paranasal sinuses and can be measured in nasally exhaled air. During humming, a marked increase in exhaled NO content has been observed. The acoustic phenomenon responsible for this evacuation of NO gas from the sinuses was analyzed. A tube model was constructed with a syringe containing NO gas attached radially. This tube was excited with an air stream modulated by a sine wave. Increased evacuation was observed whenever the syringe was not located at a pressure node of the exciting sine wave. A computer model of the system showed a good matching of observed pressure versus frequency data in the syringe resonator. The results thus suggest that the alternating pressure in the nasal cavity forces the air plug in the ostium of the paranasal sinus resonators to vibrate, thus expelling from the cavity NO gas, which is transported to free air by the exhalatory air stream. PMID- 16708929 TI - An active headrest for personal audio. AB - There is an increasing need for personal audio systems, which generate sounds that are clearly audible to one listener but are not audible to other listeners nearby. Of particular interest in this paper are listeners sitting in adjacent seats in aircraft or land vehicles. Although personal audio could then be achieved with headsets, it would be safer and more comfortable if loudspeakers in the seat headrests could be actively controlled to generate an acceptable level of acoustic isolation. In this paper a number of approaches to this problem are investigated, but the most successful involves a pair of loudspeakers on one side of the headrest, driven together to reproduce an audio signal for a listener in that seat and also to attenuate the pressures in the adjacent seat. The performance of this technique is investigated using simple analytic models and also with a practical implementation, tested in an anechoic chamber and a small room. It is found that significant attenuations, of between 5 and 25 dB, can be achieved in the crosstalk between the seats for frequencies up to about 2 kHz. PMID- 16708935 TI - Aircraft noise effects on sleep: application of the results of a large polysomnographic field study. AB - The Institute of Aerospace Medicine at the German Aerospace Center (DLR) investigated the influence of nocturnal aircraft noise on sleep in polysomnographic laboratory and field studies between 1999 and 2004. The results of the field studies were used by the Regional Council of Leipzig (Germany) for the establishment of a noise protection plan in the official approval process for the expansion of Leipzig/Halle airport. Methods and results of the DLR field study are described in detail. Special attention is given to the dose-response relationship between the maximum sound pressure level of an aircraft noise event and the probability to wake up, which was used to establish noise protection zones directly related to the effects of noise on sleep. These protection zones differ qualitatively and quantitatively from zones that are solely based on acoustical criteria. The noise protection plan for Leipzig/Halle airport is presented and substantiated: (1) on average, there should be less than one additional awakening induced by aircraft noise, (2) awakenings recalled in the morning should be avoided as much as possible, and (3) aircraft noise should interfere as little as possible with the process of falling asleep again. Issues concerned with the representativeness of the study sample are discussed. PMID- 16708941 TI - Accuracy of an acoustic location system for monitoring the position of duetting songbirds in tropical forest. AB - A field test was conducted on the accuracy of an eight-microphone acoustic location system designed to triangulate the position of duetting rufous-and-white wrens (Thryothorus rufalbus) in Costa Rica's humid evergreen forest. Eight microphones were set up in the breeding territories of 20 pairs of wrens, with an average intermicrophone distance of 75.2+/-2.6 m. The array of microphones was used to record antiphonal duets broadcast through stereo loudspeakers. The positions of the loudspeakers were then estimated by evaluating the delay with which the eight microphones recorded the broadcast sounds. Position estimates were compared to coordinates surveyed with a global-positioning system (GPS). The acoustic location system estimated the position of loudspeakers with an error of 2.82+/-0.26 m and calculated the distance between the "male" and "female" loudspeakers with an error of 2.12+/-0.42 m. Given the large range of distances between duetting birds, this relatively low level of error demonstrates that the acoustic location system is a useful tool for studying avian duets. Location error was influenced partly by the difficulties inherent in collecting high accuracy GPS coordinates of microphone positions underneath a lush tropical canopy and partly by the complicating influence of irregular topography and thick vegetation on sound transmission. PMID- 16708943 TI - Middle ear ossicles motion at hearing thresholds with air conduction and bone conduction stimulation. AB - Hearing threshold data with bone conduction and air conduction stimulation are combined with physiological and mechanical measurements of the middle ear ossicles vibration to compute the vibration level of the ossicles at threshold stimulation. By comparing the displacements of the stapes footplate with the two stimulation modalities and assuming the vibration of the stapes footplate to be the input to the cochlea when stimulation is by air conduction, the importance of middle ear ossicles inertia with bone conduction stimulation is evaluated. Given the limitations of the analysis, the results indicate that the inertia of the middle ear is not an important contribution to the perception of BC sound for frequencies below 1.5 kHz; it seems to contribute to perception of bone conducted sound between the frequencies 1.5 and 3.5 kHz. At frequencies above 4 kHz, the analysis failed since the input to the cochlea is probably not through the oval window with bone conduction stimulation. Comparison of basilar membrane vibration data verified the calculations for frequencies between 0.8 and 3.5 kHz. It was also found that the fluid flow at the round window, rather than at the oval window, reflects the stimulation of the basilar membrane with bone conduction stimulation. PMID- 16708944 TI - A geometrically nonlinear finite-element model of the cat eardrum. AB - Current finite-element (FE) models of the eardrum are limited to low pressures because of the assumption of linearity. Our objective is to investigate the effects of geometric nonlinearity in FE models of the cat eardrum with an approximately immobile malleus for pressures up to +/-2.2 kPa, which are within the range of pressures used in clinical tympanometry. Displacements computed with nonlinear models increased less than in proportion to applied pressure, similar to what is seen in measured data. In both simulations and experiments, there is a shift inferiorly in the location of maximum displacement in response to increasingly negative middle-ear pressures. Displacement patterns computed for small pressures and for large positive pressures differed from measured patterns in the position of the maximum pars-tensa displacement. Increasing the thickness of the postero-superior pars tensa in the models shifted the location of the computed maximum toward the measured location. The largest computed pars-tensa strains were mostly less than 2%, implying that a linearized material model is a reasonable approximation. Geometric nonlinearity must be considered when simulating eardrum response to high pressures because purely linear models cannot take into account the effects of changing geometry. At higher pressures, material nonlinearity may become more important. PMID- 16708945 TI - Simultaneous latency estimations for distortion product otoacoustic emissions and envelope following responses. AB - The purpose of this research was to simultaneously estimate processing delays in the cochlea and brainstem using the same acoustic stimuli. Apparent latencies were estimated from ear canal measurements of 2f1-f2 distortion product otoacoustic emissions (DPOAEs), and scalp recordings of the f2-f1 envelope following response (EFR). The stimuli were equal level tone pairs (65 dB SPL) with the upper tone f2 set at either 900 or 1800 Hz to fix the initiation site of the DPOAE and EFR. The frequency of f1 was swept continuously between frequency limits chosen to keep the EFR response between 150 and 170 Hz. The average DPOAE latencies were 9.6 and 6.2 ms for f2 =900 and 1800 Hz, and the corresponding EFR latencies were 12.4 and 8.8 ms. In a control condition, a third (suppressor) tone was added near the DPOAE response frequency to evaluate whether the potential source at fdp was contributing significantly to the measured emission. DPOAE latency is the sum of both inward and outward cochlear delays. The EFR apparent latency is the sum of inward cochlear delay and neural processing delay. Neural delay was estimated as approximately 5.3 ms for both frequencies of stimulation. PMID- 16708946 TI - Signal to noise ratio analysis of maximum length sequence deconvolution of overlapping evoked potentials. AB - In this study a general formula for the signal to noise ratio (SNR) of the maximum length sequence (MLS) deconvolution averaging is developed using the frequency domain framework of the generalized continuous loop averaging deconvolution procedure [Ozdamar and Bohorquez, J. Acoust. Soc. Am. 119, 429-438 (2006)]. This formulation takes advantage of the well known equivalency of energies in the time and frequency domains (Parseval's theorem) to show that in MLS deconvolution, SNR increases with the square root of half of the number of stimuli in the sweep. This increase is less than that of conventional averaging which is the square root of the number of sweeps averaged. Unlike arbitrary stimulus sequences that can attenuate or amplify phase unlocked noise depending on the frequency characteristics, the MLS deconvolution attenuates noise in all frequencies consistently. Furthermore, MLS and its zero-padded variations present optimal attenuation of noise at all frequencies yet they present a highly jittered stimulus sequence. In real recordings of evoked potentials, the time advantage gained by noise attenuation could be lost by the signal amplitude attenuation due to neural adaptation at high stimulus rates. PMID- 16708948 TI - Asynchrony and the grouping of vowel components: captor tones revisited. AB - Asynchrony is an important grouping cue for separating sound mixtures. A harmonic incremented in level makes a reduced contribution to vowel timbre when it begins before the other components. This contribution can be partly restored by adding a captor tone in synchrony with, and one octave above, the leading portion of the incremented harmonic [Darwin and Sutherland, Q. J. Exp. Psychol. A 36, 193-208 (1984)]. The captor is too remote to evoke adaptation in peripheral channels tuned to the incremented harmonic, and so the restoration effect is usually attributed to the grouping of the leading portion with the captor. However, results are presented that contradict this interpretation. Captor efficacy does not depend on a common onset, or harmonic relations, with the leading component. Rather, captor efficacy is influenced by frequency separation, and extends to about 1.5 oct above the leading component. Below this cutoff, the captor effect is equivalent to attenuating the leading portion of the incremented harmonic by about 6 dB. These results indicate that high-level grouping does not govern the captor effect. Instead, it is proposed that the partial restoration of the contribution of an asynchronous component to vowel timbre depends on broadband inhibition within the central auditory system. PMID- 16708947 TI - Time-course of the human medial olivocochlear reflex. AB - The time-course of the human medial olivocochlear reflex (MOCR) was measured via its suppression of stimulus-frequency otoacoustic emissions (SFOAEs) in nine ears. MOCR effects were elicited by contralateral, ipsilateral or bilateral wideband acoustic stimulation. As a first approximation, MOCR effects increased like a saturating exponential with a time constant of 277+/-62 ms, and decayed exponentially with a time constant of 159+/-54 ms. However, in ears with the highest signal-to-noise ratios (4/9), onset time constants could be separated into "fast," tau= approximately 70 ms, "medium," tau = approximately 330 ms, and "slow," tau = approximately 25 s components, and there was an overshoot in the decay like an under-damped sinusoid. Both the buildup and decay could be modeled as a second order differential equation and the differences between the buildup and decay could be accounted for by decreasing one coefficient by a factor of 2. The reflex onset and offset delays were both approximately 25 ms. Although changing elicitor level over a 20 dB SPL range produced a consistent systematic change in response amplitude, the time course did not show a consistent dependence on elictor level, nor did the time-courses of ipsilaterally, contralaterally, and bilaterally activated MOCR responses differ significantly. Given the MOCR's time-course, it is best suited to operate on acoustic changes that persist for 100's of milliseconds. PMID- 16708949 TI - Spectral loudness summation for short and long signals as a function of level. AB - The equal-loudness-ratio hypothesis (ELRH) assumes that the loudness ratio between equal-level long and short signals with the same spectrum is independent of level and spectrum. Thus, it predicts the same amount of spectral loudness summation for equal-level short and long signals. To investigate how duration and level affect spectral loudness summation, loudness functions for 10- and 1000-ms bandpass-filtered noise centered at 2 kHz were obtained for normal-hearing listeners using categorical loudness scaling. The bandwidth was varied from 200 to 6400 Hz. From the loudness functions the level difference between the 3200-Hz wide reference signal and the equally loud test signals was determined for reference levels in the range from 25 to 105 dB SPL. In addition, for two reference levels (45 and 65 dB SPL), this level difference was measured using a loudness-matching procedure. Both procedures showed similar results for the two reference levels. Over a wide level range spectral loudness summation is larger for the short than for the equal-level long signals. This is in contrast to the predictions of the ELRH. However, the data at moderate levels are consistent with the expectations of a modified ELRH which assumes a smaller loudness ratio for broadband than for narrow-band signals. PMID- 16708950 TI - Discrimination of nonlinear frequency glides. AB - Discrimination thresholds for short duration nonlinear tone glides that differed in glide rate were measured in order to determine whether cues related to rate of frequency change alone were sufficient for discrimination. Thresholds for rising and falling nonlinear glides of 50-ms and 400-ms duration, spanning three frequency excursions (0.5, 1, and 2 ERBs) at three center frequencies (0.5, 2.0, and 6.0 kHz) were measured. Results showed that glide discrimination was possible when duration and initial and final frequencies were identical. Thresholds were of a different order to those found in previous studies using linear frequency glides where endpoint frequency or duration information is available as added cues. The pattern of results was suggestive of a mechanism sensitive to spectral changes in time. Thresholds increased as the rate of transition span increased, particularly above spans of 1 ERB. The Weber fraction associated with these changes was 0.6-0.7. Overall, the results were consistent with an excitation pattern model of nonlinear glide detection that has difficulty in tracking signals with rapid frequency changes that exceed the width of an auditory filter and are of short duration. PMID- 16708951 TI - Effect of modulation maskers on the detection of second-order amplitude modulation with and without notched noise. AB - The mechanisms underlying the detection of second-order amplitude modulation (AM) were explored. The detectability of second-order AM (fixed depth for each subject) was measured for first- and second-order modulation rates of 16 and 2 Hz, respectively (slow-rate pair), and 50 and 10 Hz, respectively (fast-rate pair), with no masker, a low-band modulation masker (centered at 2 or 10 Hz), and a high-band modulation masker (centered at 16 or 50 Hz). This was done in the absence and presence of an audio-frequency notched noise centered at the carrier frequency of 4000 Hz. Both modulation maskers were "low-noise" noises, to prevent overmodulation. In the absence of notched noise, both modulation maskers impaired performance for the slow-rate pair, but only the low-band masker impaired performance for the fast-rate pair. When notched noise was present, the low-band masker had no significant effect for either rate pair and the high-band masker had an effect only for the slow-rate pair. These results suggest that second order AM detection is mediated both by an envelope distortion component at the second-order rate and by slow fluctuations in the output of a modulation filter tuned to the first-order rate. When notched noise is present, the distortion component plays little role. PMID- 16708952 TI - The influence of spectral, temporal, and interaural stimulus variations on the precedence effect. AB - The precedence effect describes phenomena that are believed to aid localization of sounds in reverberant environments. These phenomena relate to the emphasis given to the first-arriving or preceding sound. In this paper, experiments are described which study precedence using stimulus parametrizations spanning temporal, spectral, and interaural dimensions. Subjects report the sidedness of headphone stimuli comprising a source and single reflection placed symmetrically with respect to midline. Most of the experiments use long-duration noises with the onset and offset time-of-arrival differences windowed out from the combined lead and lag stimulus, thus requiring the subject to lateralize using cues in the ongoing portion of the stimuli where the lead and lag overlap completely. A similar experiment using click stimuli is included for comparison. The influence of spectral content is studied by varying either the bandwidth or the center frequency. Dependence on interaural cues is investigated by using either ITDs or IIDs to induce laterality in the individual lead and lag components. Results indicate that precedence continues into the ongoing portion of long-duration stimuli and is robust to the removal of initial onsets, to reduction of bandwidth, and to the choice of interaural cue used to induce laterality in the lead and lag. PMID- 16708953 TI - Directional loudness in an anechoic sound field, head-related transfer functions, and binaural summation. AB - The effect of sound incidence angle on loudness was investigated using real sound sources positioned in an anechoic chamber. Eight normal-hearing listeners produced loudness matches between a frontal reference location and seven sources placed at other directions, both in the horizontal and median planes. Matches were obtained via a two-interval, adaptive forced-choice (2AFC) procedure for three center frequencies (0.4, 1, and 5 kHz) and two overall levels (45 and 65 dB SPL). The results showed that loudness is not constant over sound incidence angles, with directional sensitivity varying over a range of up to 10 dB, exhibiting considerable frequency dependence, but only minor effects of overall level. The pattern of results varied substantially between subjects, but was largely accounted for by variations in individual head-related transfer functions. Modeling of binaural loudness based on the at-ear signals favored a sound-power summation model, according to which the maximum binaural gain is only 3 dB, over competing models based on larger gains, or on the summation of monaural loudness indices. PMID- 16708954 TI - Binaural detection of 500-Hz tones in broadband and in narrowband masking noise: effects of signal/masker duration and forward masking fringes. AB - NoSpi thresholds for a 500-Hz tonal signal were measured with broadband and with narrowband maskers using a single-interval adaptive matrix procedure [C. Kaernbach, J Acoust. Soc. Am. 88, 2645-2655 (1990)]. The purpose of the study was to investigate and to account for the effects on thresholds of varying the durations of the signals and maskers and the durations of forward masking fringes that preceded the occurrence of signal-plus-noise. For detection in both broadband and narrowband noise, the addition of brief forward fringes of masking noise resulted in elevations in threshold for the shortest signal durations. Longer forward fringes led to larger decreases in threshold when the masker was broadband as compared to when the masker was narrowband. The complex patterning of the data was explained by the operation of: (1) "predetection" temporal integration associated with peripheral auditory filtering; (2) duration dependent, across-frequency influences that differentially affect broadband and narrowband NoSpi thresholds, (3) "post-detection" temporal integration associated with the central binaural mechanism, and (4) consideration of the detection thresholds in terms of changes in interaural correlation rather than in terms of signal level or signal-to-noise ratio, per se. PMID- 16708955 TI - Effect of electrode configuration on psychophysical forward masking in cochlear implant listeners. AB - Bipolar stimulation has been thought to be more beneficial than monopolar stimulation for speech coding in cochlear implants, on the basis of its more restricted current flow. The present study examined whether bipolar stimulation would indeed lead to reduced channel interaction in a behavioral forward masking experiment tested in four Nucleus 24 users. The masker was fixed on one channel and three masker levels that were balanced for loudness between the configurations were chosen. As expected, masking was maximal when the masker and probe channels were spatially close and decreased as they were separated. However, overall masking patterns did not consistently demonstrate sharper tuning with bipolar stimulation than monopolar. This implies that the spatial extent of a bipolar current field is not consistently narrower than that of an equally loud monopolar stimulus; therefore, it should not be assumed that bipolar stimulation leads to reduced channel interaction. Notably, bipolar masking patterns appeared to display more variations across channels, possibly influenced more by anatomical and neural irregularities near electrode contacts than monopolar masking patterns. The present psychophysical results provide a theoretical basis regarding the widespread use (and success) of monopolar configurations by implant users. PMID- 16708956 TI - Numerical study of the effects of inferior and superior vocal fold surface angles on vocal fold pressure distributions. AB - Vocal fold geometry plays an important role in human phonation. A wide range of inferior and superior vocal fold surface angles has been shown to be present during phonation [Nanayakkara, Master's thesis, Bowling Green State University, Bowling Green, OH (2005)]. This study explored how these angles affect pressure distributions on the vocal folds, and thus how they may affect phonation. The computational code FLUENT was used to obtain pressure distributions for laminar, incompressible flow. Eighteen inferior vocal fold surface angles and nineteen superior vocal fold surface angles were selected for three specific glottal angles, uniform 0 degrees, convergent 10 degrees, and divergent 10 degrees. Minimal glottal diameter (0.01 cm), transglottal pressure (8 cm H2O), and glottal entrance radius (0.15 cm) were held constant, and the glottal exit radius was constant for each glottal angle. Results suggest that the vocal fold surface pressures are independent of the inferior and superior vocal fold surface angles realistic for human phonation. These results suggest that, in contrast to the important effects of glottal entrance and exit radii, minimal diameter, and angle on intraglottal pressures, the inferior and superior vocal fold surface angles (excluding possible interactive effects with the false vocal folds) do not have an influence on the intraglottal pressures. PMID- 16708957 TI - An experimental analysis of the pressures and flows within a driven mechanical model of phonation. AB - The production of voice is related to the flow of air through the glottis, whose time-dependent shape is defined by the motion of the vocal folds and the translaryngeal pressure. A scaled dynamically similar experimental apparatus that mimics the motion of the vocal folds was designed and built, such that both the glottal diameter and glottal angle change during a motion cycle. This motion is more realistic than in other reported dynamic models. The motion of the folds can be driven at different frequencies. The glottal flow takes place at a constant inlet pressure, mimicking the lung pressure. The transglottal pressure difference and flow rate were measured over the motion cycle. Satisfactory agreement was obtained for identical cases by numerically solving the two-dimensional, incompressible Navier-Stokes equations. Both experimental and numerical data showed that the glottal flow rate and transglottal pressure were affected by the oscillation frequency of the vocal folds. Flow visualization showed that the glottal flow patterns, which are a potential source of aero-acoustic sound, are influenced by the oscillation frequency. However, glottal flow resistance depended to a lesser extent on vocal fold oscillation frequency for the portion of the cycle when the glottis was divergent. PMID- 16708958 TI - Enhanced contrast for vowels in utterance focus: a cross-language study. AB - The present study examined several potential distinctiveness-enhancing correlates of vowels produced in utterance focus by talkers of American English, French, and German. These correlates included possible increases in vowel space size, in formant movement within individual vowels, and in duration variance among vowels. Each language group enhanced the distinctiveness of vowels in [+focus] context but used somewhat differing means to achieve this. All three groups used spectral differences, but only German talkers used durational differences, to enhance distinctiveness. The results suggest that the amount of distinctiveness enhancement of a vowel property in [+focus] context is positively related to the between-category variation of that property in [-focus] context. Thus, consistent with the theory of adaptive dispersion, utterance clarity appears to vary directly with information content. PMID- 16708959 TI - Comparing the rhythm and melody of speech and music: the case of British English and French. AB - For over half a century, musicologists and linguists have suggested that the prosody of a culture's native language is reflected in the rhythms and melodies of its instrumental music. Testing this idea requires quantitative methods for comparing musical and spoken rhythm and melody. This study applies such methods to the speech and music of England and France. The results reveal that music reflects patterns of durational contrast between successive vowels in spoken sentences, as well as patterns of pitch interval variability in speech. The methods presented here are suitable for studying speech-music relations in a broad range of cultures. PMID- 16708960 TI - Language redundancy predicts syllabic duration and the spectral characteristics of vocalic syllable nuclei. AB - The language redundancy of a syllable, measured by its predictability given its context and inherent frequency, has been shown to have a strong inverse relationship with syllabic duration. This relationship is predicted by the smooth signal redundancy hypothesis, which proposes that robust communication in a noisy environment can be achieved with an inverse relationship between language redundancy and the predictability given acoustic observations (acoustic redundancy). A general version of the hypothesis predicts similar relationships between the spectral characteristics of speech and language redundancy. However, investigating this claim is hampered by difficulties in measuring the spectral characteristics of speech within large conversational corpora, and difficulties in forming models of acoustic redundancy based on these spectral characteristics. This paper addresses these difficulties by testing the smooth signal redundancy hypothesis with a very high-quality corpus collected for speech synthesis, and presents both durational and spectral data from vowel nuclei on a vowel-by-vowel basis. Results confirm the duration/language redundancy results achieved in previous work, and show a significant relationship between language redundancy factors and the first two formants, although these results vary considerably by vowel. In general, however, vowels show increased centralization with increased language redundancy. PMID- 16708961 TI - Cue weighting in auditory categorization: implications for first and second language acquisition. AB - The ability to integrate and weight information across dimensions is central to perception and is particularly important for speech categorization. The present experiments investigate cue weighting by training participants to categorize sounds drawn from a two-dimensional acoustic space defined by the center frequency (CF) and modulation frequency (MF) of frequency-modulated sine waves. These dimensions were psychophysically matched to be equally discriminable and, in the first experiment, were equally informative for accurate categorization. Nevertheless, listeners' category responses reflected a bias for use of CF. This bias remained even when the informativeness of CF was decreased by shifting distributions to create more overlap in CF. A reversal of weighting (MF over CF) was obtained when distribution variance was increased for CF. These results demonstrate that even when equally informative and discriminable, acoustic cues are not necessarily equally weighted in categorization; listeners exhibit biases when integrating multiple acoustic dimensions. Moreover, changes in weighting strategies can be affected by changes in input distribution parameters. This methodology provides potential insights into acquisition of speech sound categories, particularly second language categories. One implication is that ineffective cue weighting strategies for phonetic categories may be alleviated by manipulating variance of uninformative dimensions in training stimuli. PMID- 16708962 TI - Early learners' discrimination of second-language vowels. AB - It is uncertain from previous research to what extent the perceptual system retains plasticity after attunement to the native language (L1) sound system. This study evaluated second-language (L2) vowel discrimination by individuals who began learning the L2 as children ("early learners"). Experiment 1 identified procedures that lowered discrimination scores for foreign vowel contrasts in an AXB test (with three physically different stimuli per trial, where "X" was drawn from the same vowel category as "A" or "B"). Experiment 2 examined the AXB discrimination of English vowels by native Spanish early learners and monolingual speakers of Spanish and English (20 per group) at interstimulus intervals (ISIs) of 1000 and 0 ms. The Spanish monolinguals obtained near-chance scores for three difficult vowel contrasts, presumably because they did not perceive the vowels as distinct phonemes and because the experimental design hindered low-level encoding strategies. Like the English monolinguals, the early learners obtained high scores, indicating they had shown considerable perceptual learning. However, statistically significant differences between early learners and English monolinguals for two of three difficult contrasts at the 0-ms ISI suggested that their underlying perceptual systems were not identical. Implications for claims regarding perceptual plasticity following L1 attunement are discussed. PMID- 16708963 TI - Phonological versus phonetic cues in native and non-native listening: Korean and Dutch listeners' perception of Dutch and English consonants. AB - We investigated how listeners of two unrelated languages, Korean and Dutch, process phonologically viable and nonviable consonants spoken in Dutch and American English. To Korean listeners, released final stops are nonviable because word-final stops in Korean are never released in words spoken in isolation, but to Dutch listeners, unreleased word-final stops are nonviable because word-final stops in Dutch are generally released in words spoken in isolation. Two phoneme monitoring experiments showed a phonological effect on both Dutch and English stimuli: Korean listeners detected the unreleased stops more rapidly whereas Dutch listeners detected the released stops more rapidly and/or more accurately. The Koreans, however, detected released stops more accurately than unreleased stops, but only in the non-native language they were familiar with (English). The results suggest that, in non-native speech perception, phonological legitimacy in the native language can be more important than the richness of phonetic information, though familiarity with phonetic detail in the non-native language can also improve listening performance. PMID- 16708964 TI - Speech perception in gated noise: the effects of temporal resolution. AB - Previous investigations have suggested that hearing-impaired (HI) listeners have reduced masking release (MR) compared to normal hearing listeners (NH) when they listen in modulated noise. The current study examined the following questions that have not been clearly answered: First, when HI listeners are amplified so that their performance is equal to that of NH listeners in quiet and in steady noise, do HI listeners still show reduced MR with modulated noise when compared to NH listeners? Second, is the masking release the same for sentences and CV syllables? Third, does forward masking significantly contribute to the variability in performance among HI listeners? To compensate for reduced hearing sensitivity for HI listeners, the spectrum levels of both speech and noise were adjusted based on the individual hearing loss. There was no significant difference between the performance of NH listeners and that of HI listeners in steady noise and in quiet. However, the amount of MR for sentences and for CV syllables was significantly reduced for HI listeners. For sentence recognition, the amount of MR seemed to be more related to hearing sensitivity for low-to-mid frequencies than to forward masking. In contrast, forward masking thresholds appear to be a major contributor to the amount of MR for syllable recognition. PMID- 16708965 TI - A rule-based emotion-dependent feature extraction method for emotion analysis from speech. AB - This paper presents a rule-based method to determine emotion-dependent features, which are defined from high-level features derived from the statistical measurements of prosodic parameters of speech. Emotion-dependent features are selected from high-level features using extraction rules. The ratio of emotional expression similarity between two speakers is defined by calculating the number and values of the emotion-dependent features that are present for the two speakers being compared. Emotional speech from Interface databases is used for evaluation of the proposed method, which was used to analyze emotional speech from five male and four female speakers in order to find any similarities and differences among individual speakers. The speakers are actors that have interpreted six emotions in four different languages. The results show that all the speakers share some universal signs regarding certain emotion-dependent features of emotional expression. Further analysis revealed that almost all speakers in the analysis used unique sets of emotion-dependent features and each speaker used unique values for the defined emotion-dependent features. The comparison among speakers shows that the expressed emotions can be analyzed according to two criteria. The first criterion is a defined set of emotion dependent features and the second is an emotion-dependent feature value. PMID- 16708968 TI - Using self-organizing maps to recognize acoustic units associated with information content in animal vocalizations. AB - Kohonen self-organizing neural networks, also called self-organizing maps (SOMs), have been used successfully to recognize human phonemes and in this way to aid in human speech recognition. This paper describes how SOMS also can be used to associate specific information content with animal vocalizations. A SOM was used to identify acoustic units in Gunnison's prairie dog alarm calls that were vocalized in the presence of three different predator species. Some of these acoustic units and their combinations were found exclusively in the alarm calls associated with a particular predator species and were used to associate predator species information with individual alarm calls. This methodology allowed individual alarm calls to be classified by predator species with an average of 91% accuracy. Furthermore, the topological structure of the SOM used in these experiments provided additional insights about the acoustic units and their combinations that were used to classify the target alarm calls. An important benefit of the methodology developed in this paper is that it could be used to search for groups of sounds associated with information content for any animal whose vocalizations are composed of multiple simultaneous frequency components. PMID- 16708967 TI - Measurements of the anisotropy of ultrasonic attenuation in freshly excised myocardium. AB - Echocardiography requires imaging of the heart with sound propagating at varying angles relative to the predominant direction of the myofibers. The degree of anisotropy of attenuation can significantly influence ultrasonic imaging and tissue characterization measurements in vivo. This study quantifies the anisotropy of attenuation of freshly excised myocardium at frequencies typical of echocardiographic imaging. Results show a significantly larger anisotropy than previously reported in specimens of locally unidirectional myofibers. Through transmission radio frequency-based measurements were performed on specimens from 12 ovine and 12 bovine hearts. Although ovine hearts are closer in size to human, the larger size of bovine hearts offers the potential for specimens in which myofibers are more nearly unidirectionally aligned. The attenuation coefficient increased approximately linearly with frequency. The mean slope of attenuation with frequency was 3-4 times larger for propagation parallel than for perpendicular to the myofibers. At perpendicular insonification, slopes between ovine and bovine myocardium were approximately equal. However, attenuation in bovine specimens was larger for angles approaching parallel. The difference in results for parallel appears consistent with what might be expected from increased myofiber curvature associated with smaller lamb hearts. Quantitative knowledge of anisotropy of attenuation may be useful in understanding mechanisms underlying the interaction of ultrasound with myocardium. PMID- 16708969 TI - Tissue deformation induced by radiation force from Gaussian transducers. AB - Imaging techniques based upon the tissue mechanical response to an acoustic radiation force are being actively researched. In this paper a model for predicting steady-state tissue displacement induced by a radiation force arising from the absorption of Gaussian ultrasound beams is presented. A simple analytic expression is derived that agrees closely with the numerical quadrature of the displacement convolution integrals. The analytic result reveals the dependence of the steady-state axial displacement upon the operational parameters, e.g., an inverse proportional relationship to the tissue shear modulus. The derivation requires that the transducer radius be small compared to the focal length, but accurate results were obtained for transducer radii comparable to the focal length. Favorable comparisons with displacement predictions for non-Gaussian transducers indicate that the theory is also useful for a broader range of transducer intensity profiles. PMID- 16708970 TI - Acoustic structures in the alarm calls of Gunnison's prairie dogs. AB - Acoustic structures of sound in Gunnison's prairie dog alarm calls are described, showing how these acoustic structures may encode information about three different predator species (red-tailed hawk-Buteo jamaicensis; domestic dog-Canis familaris; and coyote-Canis latrans). By dividing each alarm call into 25 equal sized partitions and using resonant frequencies within each partition, commonly occurring acoustic structures were identified as components of alarm calls for the three predators. Although most of the acoustic structures appeared in alarm calls elicited by all three predator species, the frequency of occurrence of these acoustic structures varied among the alarm calls for the different predators, suggesting that these structures encode identifying information for each of the predators. A classification analysis of alarm calls elicited by each of the three predators showed that acoustic structures could correctly classify 67% of the calls elicited by domestic dogs, 73% of the calls elicited by coyotes, and 99% of the calls elicited by red-tailed hawks. The different distributions of acoustic structures associated with alarm calls for the three predator species suggest a duality of function, one of the design elements of language listed by Hockett [in Animal Sounds and Communication, edited by W. E. Lanyon and W. N. Tavolga (American Institute of Biological Sciences, Washington, DC, 1960), pp. 392-430]. PMID- 16708971 TI - Using an artificial neural network to classify black-capped chickadee (Poecile atricapillus) call note types. AB - The "chick-a-dee" call of the black-capped chickadee (Poecile atricapillus) contains four note types, A, B, C, and D that have important functional roles. This provides strong motivation for studying the classification of acoustic components of the call into different note types. In this paper, the spectrograms from a sample of A, B, and C notes (370 in total) were each described as a set of 9 summary features. An artificial neural network was trained to identify note type on the basis of these features, and was able to obtain better than 98% accuracy. An internal analysis of this network revealed a distributed code in which different hidden units generated high activities to different subsets of notes. By combining these different sensitivities, the network could discriminate all three types of notes. The performance of this network was compared to a discriminant analysis of the same data. This analysis also achieved a high level of performance (95%). A comparison between the two approaches revealed some striking similarities, but also some intriguing differences. These results are discussed in terms of two related issues: developing a research tool for note classification, and developing a theory of how birds themselves might classify notes. PMID- 16708972 TI - Nonconstant quality of auditory filters in the porpoises, Phocoena phocoena and Neophocaena phocaenoides (Cetacea, Phocoenidae). AB - Simultaneous tone-tone masking in conjunction with the envelope-following response (EFR) recording was used to obtain tuning curves in porpoises Phocoena phocoena and Neophocaena phocaenoides asiaeorientalis. The EFR was evoked by amplitude-modulated probes with a modulation rate of 1000 Hz and carrier frequencies from 22.5 to 140 kHz. Equivalent rectangular quality QERB of the obtained tuning curves varied from 8.3-8.6 at lower (22.5-32 kHz) probe frequencies to 44.8-47.4 at high (128-140 kHz) frequencies. The QERB dependence on probe frequency could be approximated by regression lines with a slope of 0.83 to 0.86 in log-log scale, which corresponded to almost frequency-proportional quality and almost constant bandwidth of 3-4 kHz. Thus, the frequency representation in the porpoise auditory system is much closer to a constant bandwidth rather that to a constant-quality manner. PMID- 16708973 TI - Comparison of in-air evoked potential and underwater behavioral hearing thresholds in four bottlenose dolphins (Tursiops truncatus). AB - Traditional behavioral techniques for hearing assessment in marine mammals are limited by the time and access required to train subjects. Electrophysiological methods, where passive electrodes are used to measure auditory evoked potentials (AEPs), are attractive alternatives to behavioral techniques; however, there have been few attempts to compare AEP and behavioral results for the same subject. In this study, behavioral and AEP hearing thresholds were compared in four bottlenose dolphins. AEP thresholds were measured in-air using a piezoelectric sound projector embedded in a suction cup to deliver amplitude modulated tones to the dolphin through the lower jaw. Evoked potentials were recorded noninvasively using surface electrodes. Adaptive procedures allowed AEP hearing thresholds to be estimated from 10 to 150 kHz in a single ear in about 45 min. Behavioral thresholds were measured in a quiet pool and in San Diego Bay. AEP and behavioral threshold estimates agreed closely as to the upper cutoff frequency beyond which thresholds increased sharply. AEP thresholds were strongly correlated with pool behavioral thresholds across the range of hearing; differences between AEP and pool behavioral thresholds increased with threshold magnitude and ranged from 0 to + 18 dB. PMID- 16708974 TI - AARP skews younger. PMID- 16708975 TI - How the VA healed itself. PMID- 16708976 TI - The ultimate Bing diet. PMID- 16708977 TI - The club. One nurse's role in a social day care program. AB - Social day care is an important component in the health care continuum for older adults. At present there are no regulations mandating staff composition, therefore each program may have a different staff configuration. One such social day program, The Club, provides services to individuals with dementia and has had a nurse as part of its interdisciplinary team since its inception in 1984. Nursing in a social day care environment is challenging and draws on all of one's nursing skills. The nurse is an important member of the day care staff: The nurse helps individuals with dementia achieve success in the program and assists the caregiver through education and counseling to maintain the person in the community. This Spotlight features an overview of the multiple and interdisciplinary roles Ellen Tolle, RN, MPS plays as Program Director at The Club. These include the nurse as administrator, clinician, educator, and counselor in a dementia-specific social day program. PMID- 16708978 TI - Respect and dignity: Enduring concepts, enduring challenges. PMID- 16708979 TI - The medical futility debate: treatment at any cost? PMID- 16708980 TI - Caring for dying patients and those facing death in an acute-care hospital in Singapore: a nurse's perspective. AB - Nurses are at the forefront of caring for dying patients in hospices, nursing homes, acute-care hospitals, and patients' homes. This study was conducted to explore the emotional and practical experience as well as attitudes of nurses caring for the dying and the deceased in an acute-care hospital in Singapore. The authors explored differences in opinion among the various ethnic groups (Malay, Chinese, Indian, Filipino) that make up the nurse population in this particular hospital. A structured questionnaire was self-administered by the participants to explore nurses' views, opinions, and experiences in caring for dying patients from a quantitative aspect. It specifically addressed attitudinal, ethical, and communicational issues involved in caring for dying patients. Where applicable, a Likert scale ranging from Always to Never or from Strongly Agree to Strongly Disagree was used. One hundred and eighty of 246 questionnaires were returned (73%). Most respondents never or only occasionally felt uncomfortable caring for dying patients, and felt it reminded them of their own mortality, made them treasure life more, and made them ask questions about life and death. In this descriptive study, some difference in attitudes and views among the various ethnic groups was observed. PMID- 16708981 TI - Careful hand feeding: a reasonable alternative to PEG tube placement in individuals with dementia. AB - Individuals with dementia inevitably experience decreased oral intake when they are no longer able to recognize food, feed themselves independently, or experience swallowing difficulties. There is mounting evidence that routine insertion of a percutaneous endoscopic gastrostomy (PEG) does not significantly affect survival, nor does it reduce the risk of aspiration pneumonia and pressure ulcer formation. Although there is relatively little published on assisted oral feeding, its advantages include allowing the patient to enjoy the gratification of eating and socialization that accompanies mealtime, as well as avoiding restraint use that often accompanies PEG placement. Nurses can play a pivotal role in assessment of feeding difficulties, implementation of innovative mealtime programs, educating families about specific strategies to optimize oral intake, and participating in research to evaluate patient outcomes. PMID- 16708982 TI - Positive gain: rethinking the outcomes of dependency. AB - While independent behavior by older adults is routinely encouraged, some individuals can benefit from engaging in dependent behavior. In traditional views of dependency, dependent behaviors are considered maladaptive and dysfunctional. The purpose of this article is to consider the interactions between health professionals and older patients, the ways that dependent behaviors can lead to both positive outcomes for older adults, as well as serve as barriers to self care and examine the role of nurses and institutions in interactions with older adults. An improved understanding of dependency and its relevance to nursing practice will contribute to a heightened awareness of the control dynamics occurring in everyday nursing care activities. An understanding of potential gains attained through dependency can add another dimension to assessment of a patient's progress and capability for present and future self-care. PMID- 16708983 TI - Care with dignity: a concept analysis. AB - The attribute of dignity depends on the perspective of the individual and can be affected by interactions with others. Dignity involves the day-to-day choices by which individuals create who they will become and how they will be perceived in the context of human culture. Dignified care requires a caregiver-patient relationship that values autonomy, individual diversity, truth, justice, rights, and responsibilities. Because authoritative practice does not acknowledge the rights and dignity of others, those who have power must constrain themselves and use caution in applying judgement. PMID- 16708984 TI - Perceptions of empowerment and respect: effect on nurses' organizational commitment in nursing homes. AB - This study tested Kanter's theory of structural empowerment in a sample of nurses employed in nursing homes throughout Ontario by examining relationships between nurses' perceptions of structural and psychological empowerment, respect, and organizational commitment. A random sample of 79 RNs (response rate 64%) and 75 RPNs (response rate 60%) were used. Both groups reported moderate levels of empowerment, respect, and commitment. RNs perceived higher levels of empowerment and respect than RPNs. Access to opportunity was the most empowering factor for nurses in this study, and access to resources the least empowering. Structural empowerment, psychological empowerment, and respect explained 48% of the variance in affective commitment for RNs and 40% for RPNs. Results of this study provide support for the use of Kanter's theory in LTC nurse populations. PMID- 16708985 TI - [To learn by heart]. PMID- 16708986 TI - Basic research and the practice of medicine. PMID- 16708987 TI - Cardiologists are living through exciting times. The example of postconditioning to protect the human heart during revascularization. AB - The exciting work the group of Jennings, published in Circulation in 1986, has established a new and powerful form of cardioprotection, whereby exposure to short bouts of repeated ischemia interspersed by reperfusion protects from a subsequent prolonged and potentially very serious major ischemic attack. Similar protection by postconditioning can be achieved by short bursts of ischemia just after the onset of mechanical reperfusion in acute myocardial infarction. Of interest, it has taken almost 20 years of intense work to bring all the important preconditioning ideas to clinical reality in the form of postconditioning. Once cardiologists are fully awoken to the idea of attacking reperfusion induced myocardial cell death, then this area will be set to develop as a major and novel target in the therapy of acute myocardial infarction. PMID- 16708988 TI - [Postconditioning: lethal reperfusion injury as a therapeutic target]. AB - Acute myocardial infarction is the leading cause of morbidity and mortality in western countries. Ischemic postconditioning, that consists of repeated brief episodes of ischemia-reperfusion performed just after reflow following a prolonged ischemic insult, dramatically reduces infarct size. Recent data indicate that it might involve the activation of the PI3-kinase-Akt-eNOS signalling pathway and inhibition of the opening of the permeability transition pore. A recent clinical study demonstrated that postconditioning protects the human heart. Further research is needed to find new pharmacological agents that would mimick postconditioning in order to treat all patients with ongoing acute myocardial infarction. PMID- 16708989 TI - [Hypoxic preconditioning: role of transcription factor HIF-1alpha]. AB - The delayed form of myocardial preconditioning is of particular interest because of its large window of protection. It involves many signalisation pathways who, along with transcription factors, activate cardioprotective genes. Amongst the latter, the hypoxia inducible factor 1 (HIF-1) whose a subunit is stabilized by hypoxia, appears to play a pivotal role in the delayed preconditioning induced by hypoxia. The stabilisation of HIF-1alpha by inhibitors of prolyl-4-hydroxylases, the enzymes responsible for its degradation in normoxia, reproduces the cardioprotective effects of hypoxia. These enzymes represent promising therapeutic targets for the treatment of various cardiovascular diseases. PMID- 16708990 TI - [The toxic aggregation of proteins: a kind of "molecular delinquency" actively fought in the cell by molecular chaperones and proteases]. AB - Under various stresses, mutation-sensitised proteins may spontaneously convert into inactive, aggregation-prone structures, which may be cytotoxic and infectious. In the cell, this new kind of "molecular criminality" is actively fought against by a network of molecular chaperones that can specifically identify, isolate and unfold damaged (delinquent) proteins and favour their subsequent native refolding. Irreversibly damaged molecules unable to natively refold are preferentially "executed" and recycled by proteases. Failing that, they are "imprisoned" within compact amyloids, or "evicted" from the cell. Thus, striking parallels, although of questionable ethical value, exist between protein and human criminality, and between the cellular and social responses to these different types of criminality. Fundamental differences also exist. Whereas programmed death (apoptosis) is the preferred solution chosen by aged and aggregation-stressed cells, collective suicide is seldom an option chosen by lawless human societies. More significantly, there is no clear cellular equivalent for the role of the family and the education system, which are so essential to the proper shaping of functional individuals in the society, and give rise to humanism, that favours crime prevention, reeducation and reinsertion programs over capital punishment. To the cardiologist and transplantation surgeon, the interest of molecular chaperones, in particular of Hsp70, Hsp90 and Hsp27, lays in their ability to inhibit the signalling pathway of programmed cell death. Their induction before and during ischemia, by various treatments and drugs could significantly reduce damages from the post ischemic reperfusion of organs. PMID- 16708991 TI - Physiopathology of the embryonic heart (with special emphasis on hypoxia and reoxygenation). AB - The adaptative response of the developing heart to adverse intrauterine environment such as reduced O2 delivery can result in alteration of gene expression with short- and long-term consequences including adult cardiovascular diseases. The tolerance of the developing heart of acute or chronic oxygen deprivation, its capacity to recover during reperfusion and the mechanisms involved in reoxygenation injury are still under debate. Indeed, the pattern of response of the immature myocardium to hypoxia-reoxygenation differs from that of the adult. This review deals with the structural and metabolic characteristics of the embryonic heart and the functional consequences of hypoxia and reoxygenation. The relative contribution of calcium and sodium overload, pH disturbances and oxidant stress to the hypoxia-induced cardiac dysfunction is examined, as well as various cellular signaling pathways (e.g. MAP kinases) involved in cell survival or death. In the context of the recent advances in developmental cardiology and fetal cardiac surgery, a better understanding of the physiopathology of the stressed developing heart is required. PMID- 16708992 TI - [Development of cardiac physiopathological models from cultured cardiomyocytes]. AB - The cultures of neonatal rat cardiomyocytes represent a very useful tool for the observation and the understanding of the cellular aspects of the electrophysiological, contractile, morphological, metabolic and molecular properties of the myocardium. This model is characterized by a homogeneous population of cardiac muscular cells and by vast possibilities of control of the chemical and physical environment of the cells, allowing the in vitro mimicry of a wide range of cardiac pathological situations. The cardiomyocyte cultures are thus suited to very varied experimental protocols, allowing multiparametric analysis of the cardiocellular effects of different stress such as hypoxia reoxygenation, of ischemia-reperfusion, of the free radical attack and of thermal shock. These investigations can be combined with the study of the effects and of the cytotoxicity of pharmacological agents, not limited to the putatively cardioactive drugs. The present review proposes an outline of the procedures for the isolation, the culture and the use of neonatal cardiomyocytes. To illustrate the potentialities of this preparation, we describe more specifically the protocols and the various consequences at the cellular scale of an in vitro model of myocardial ischemia reperfusion. PMID- 16708993 TI - [Diabetes and peripheral arterial occlusive disease: therapeutic potential and pro-angiogenic strategies]. AB - Cardiovascular complications are the leading cause of morbidity and mortality in patients with diabetes mellitus; up to 80% of deaths in patients with diabetes are closely associated with vascular disease. The ability of the organism to form a collateral network of blood vessels constitutes an important response to vascular occlusive disease and determines to a large part the clinical consequences and severity of tissue ischemia. The development of new vessels is significantly reduced in diabetic patients with coronary or peripheral artery disease. This probably contributes to the severe course of limb ischemia in diabetic patients, in which peripheral artery disease often results in foot ulceration and lower extremity amputation. Diabetic retinopathy remains one of the major causes of acquired blindness in developed nations. This is true despite the development of laser treatment, which can prevent blindness in the majority of those who develop macular edema or proliferative diabetic retinopathy. The hallmark of diabetic retinopathy is the lack of microvessels in the macula, leading to hypoxia, associated with peripheral retinal neovascularization that may ultimately cause severe vitreous cavity bleeding and/or retinal detachment. The factors that stimulate retinal blood vessel growth have not been fully defined, but there is accumulating evidence that the renin-angiotensin-bradykinin system may be involved in a number of retinal vascular disorders, including retinopathy of prematurity and proliferative diabetic retinopathy. Only a few studies have specifically evaluated the effect of diabetes on angiogenesis in ischemic vascular disease and in the retina. Moreover, the mechanisms by which diabetes could both limit the formation of new blood vessels in most organs and simultaneously induce proliferative diabetic retinopathy remain largely undefined. In the present review, we aimed to briefly describe the main molecular mechanisms involved in the ischemia-induced angiogenesis, and their alterations in diabetes. Possible therapeutic strategies to restore angiogenesis in diabetic patients are also listed. PMID- 16708994 TI - [Preventing the cardiotoxic effects of anthracyclins. From basic concepts to clinical data]. AB - The chronic cardiotoxicity of the cytotoxic agents such as anthracyclines is one of the main factors, which limits their prolonged use. Clinically, this cardiotoxicity results in a cardiomyopathy with irreversible congestive heart failure, with high mortality. The molecular mechanisms, which could explain this cardiac toxicity, are complex but it seems distinct from the anticancer mechanism. Several hypotheses were advanced but it appears that the induction of an oxidative stress within myocardial tissue constitutes the common denominator. The prevention of this cardiotoxicity lies on:--a rigorous cardiac monitoring- the use of anthracyclines analogues with lower cardiotoxicity,--modifications of the protocols of administration. The myocardial protection, with cardioprotective agents targeting oxidative stress during chemotherapy would be of great interest for an optimal use of the anthracyclines. PMID- 16708995 TI - Chemogenomics in drug discovery. AB - Chemogenomics is a new strategy in drug discovery which, in principle, searches for all molecules that are capable of interacting with any biological target. Because of the almost infinite number of drug-like organic molecules, this is an impossible task. Therefore chemogenomics has been defined as the investigation of classes of compounds (libraries) against families of functionally related proteins. In this definition, chemogenomics deals with the systematic analysis of chemical-biological interactions. Congeneric series of chemical analogs are probes to investigate their action on specific target classes, e.g., GPCRs, kinases, phosphodiesterases, ion channels, serine proteases, and others. Whereas such a strategy developed in pharmaceutical industry almost 20 years ago, it is now more systematically applied in the search for target- and subtype-specific ligands. The term "privileged structures" has been defined for scaffolds, such as the benzodiazepines, which very often produce biologically active analogs in a target family, in this case in the class of G-protein-coupled receptors. The SOSA approach is a strategy to modify the selectivity of biologically active compounds, generating new drug candidates from the side activities of therapeutically used drugs. PMID- 16708996 TI - Chemogenomics strategies for G-protein coupled receptor hit finding. AB - Targeting protein superfamilies via chemogenomics is based on a similarity clustering of gene sequences and molecular structures of ligands. Both target and ligand clusters are linked by generating binding affinity profiles of chemotypes vs a target panel. The application of this multidimensional similarity paradigm will be described in the context of Lead Generation to identify novel chemical hit classes for G-protein coupled receptors. PMID- 16708997 TI - Chemogenomics approaches to G-protein coupled receptor lead finding. AB - G-protein coupled receptors (GPCRs) are promising targets for the discovery of novel drugs. In order to identify novel chemical series, high-throughput screening (HTS) is often complemented by rational chemogenomics lead finding approaches. We have compiled a GPCR directed screening set by ligand-based virtual screening of our corporate compound database. This set of compounds is supplemented with novel libraries synthesized around proprietary scaffolds. These target-directed libraries are designed using the knowledge of privileged fragments and pharmacophores to address specific GPCR subfamilies (e.g., purinergic or chemokine-binding GPCRs). Experimental testing of the GPCR collection has provided novel chemical series for several GPCR targets including the adenosine A1, the P2Y12, and the chemokine CCR1 receptor. In addition, GPCR sequence motifs linked to the recognition of GPCR ligands (termed chemoprints) are identified using homology modeling, molecular docking, and experimental profiling. These chemoprints can support the design and synthesis of compound libraries tailor-made for a novel GPCR target. PMID- 16708998 TI - Diversity oriented synthesis: a challenge for synthetic chemists. AB - This article covers the diversity-oriented synthesis (DOS) of small molecules in order to generate a collection of pure compounds that are attractive for lead generation in a phenotypic, high-throughput screening approach useful for chemical genetics and drug discovery programmes. Nature synthesizes a rich structural diversity of small molecules, however, unfortunately, there are some disadvantages with using natural product sources for diverse small-molecule discovery. Nevertheless we have a lot to learn from nature. The efficient chemical synthesis of structural diversity (and complexity) is the aim of DOS. Highlights of this article include a discussion of nature's and synthetic chemists' strategies to obtain structural diversity and an analysis of molecular descriptors used to classify compounds. The assessment of how successful one diversity-oriented synthesis is vs another is subjective; therefore we use freely available software (www.cheminformatics.org/diversity) to assess structural diversity in any combinatorial synthesis. PMID- 16708999 TI - Probing protein function with small molecules. AB - The interface of chemistry and biology offers many opportunities to explore different aspects of cell biology. The emerging field of chemical genetics is providing the chemical means to understand biological systems not easily accessible using classical genetic manipulations. In this article, we will discuss how natural product mode of action studies and novel bio-organic manipulation of intracellular protein levels are proving useful in the exploration of cell biology. PMID- 16709001 TI - Protein structure similarity clustering and natural product structure as guiding principles for chemical genomics. AB - The majority of all proteins are modularly built from a limited set of approximately 1,000 structural domains. The knowledge of a common protein fold topology in the ligand-sensing cores of protein domains can be exploited for the design of small-molecule libraries in the development of inhibitors and ligands. Thus, a novel strategy of clustering protein domain cores based exclusively on structure similarity considerations (protein structure similarity clustering, PSSC) has been successfully applied to the development of small-molecule inhibitors of acetylcholinesterase and the 11beta-hydroxysteroid dehydrogenases based on the structure of a naturally occurring Cdc25 inhibitor. The efficiency of making use of the scaffolds of natural products as biologically prevalidated starting points for the design of compound libraries is further highlighted by the development of benzopyran-based FXR ligands. PMID- 16709000 TI - Tamoxifen-based probes for the study of estrogen receptor-mediated transcription. AB - The nuclear receptors are ideal targets to control the expression of specific genes with small molecules. Estrogen receptor can activate or repress transcription though a number of different pathways. As part of an effort to develop reagents that selectively target specific transcriptional regulatory pathways, analogs of 4-hydroxytamoxifen were synthesized with variations in the basic side chain. In vitro binding assays and cell-based luciferase reporter gene assays confirm that all the derivatives have high affinity for the receptor and high potency at repressing direct estrogen receptor-mediated transcription. PMID- 16709002 TI - Tackling the chemogenomic space by novel screening technologies. AB - Drug discovery in the chemogenomic space has seen some tremendous changes over the last decade. Compared to previous times, not only the number of available chemical compounds for screening, but also the number of molecular targets used for screening has increased significantly. This has triggered the need for very fast, efficient, and effective novel readout technologies for compound testing. Novartis has developed two novel high-throughput screening (HTS) technologies for that purpose--NanoScreen and SpeedScreen. NanoScreen is a highly miniaturized and fully automated HTS/uHTS test system with confocal single-molecule as well as non confocal detection capabilities and is used for functional screening in the range of 1-5 microl per sample. The integration of the single-molecule readout technologies into the system enables highly sophisticated biochemical test systems with multi-parameter readout for very high data quality. SpeedScreen is a highly miniaturized and automated screening system for high-throughput affinity selection of compounds. In practice, pools of compounds are incubated with the target protein and the unbound chemical compounds are removed from the target compound complex via very fast, multiparallel size-exclusion-chromatography. The holoenzyme is disintegrated and analyzed via microbore reversed-phase high performance liquid chromatography (microbore RP-HPLC). Both systems have been developed and implemented with great success at the Novartis Lead Discovery Center (LDC) in Basel. These technologies have enabled us to access targets that would otherwise not have been possible, e.g., very expensive targets, "orphan" drug targets, or targets that are "non-tractable" by conventional screening technologies. Taken together, these novel screening technologies enable novel approaches for chemogenomic research that would have not been possible in the past. PMID- 16709003 TI - When parents are the threat. PMID- 16709004 TI - Your drug was discontinued. PMID- 16709005 TI - [Drug-induced renal calculi]. AB - Drug-induced renal calculi represent 1-2% of all renal calculi. They include two categories: those resulting from the urinary crystallisation of a highly excreted, poorly soluble drug or metabolite, and those due to the metabolic effects of a drug. Indinavir, used in HIV-infected patients, sulfonamides, especially sulfadiazine, and triamterene, which is less prescribed today, are the most frequent. Besides these drugs, about twenty other molecules, among them silicate-containing drugs and some antibiotics have been reported in patients receiving high doses or long-term treatments. Calculi analysis by physical methods such as infrared spectroscopy or x-ray diffraction can demonstrate the presence of the drug or its metabolites inside the calculi. In those calculi due to the metabolic effects of a drug, diagnosis relies on both stone analysis and clinical inquiry. Incidence of such calculi is probably underestimated, especially those due to calcium/vitamin D supplements or carbonic anhydrase inhibitors. Drug-induced calculi occur more often during high-dose or long term treatments, but there are also patient-related risk factors in relation to urine pH, urine output and other parameters, which can provide a basis for preventive treatment of such calculi. A better knowledge of these lithogenic complications of treatments and of solubility characteristics of drugs should reduce the incidence of drug-induced nephrolithiasis, especially in patients with identified risk factors. PMID- 16709006 TI - [Imaging of urinary lithiasis: "all in one"]. AB - Urinary lithiasis is very common among the general population, with a high prevalence level. In rich countries it is mainly located inside the upper urinary tract. Helical CToperated with newer devices is the most accurate modality to provide all needed information: diagnosis of stone without contrast medium injection, morphology (size, number) and localization, diagnosis of urinary obstruction, urinary tract aspect and all kind of differential diagnosis in emergency. It must be used during follow up to diagnose residual fragments. Multiplanar reconstructions are essential for the clinicians; but diagnosis is interpreted by scrolling axial views with dynamic analysis on computer screen. Low-dose helical CT is today available. Helical CT provides an "all-in-one" examination. It should soon replace combined plain film-ultrasonography performed in an emergency context of renal colic and intravenous urography for pre- and post-treatment assessments. PMID- 16709007 TI - [Management of T1G3 tumours of the bladder]. AB - T1G3 tumours are the most aggressive superficial tumours of the bladder, with a high risk of recurrence and progression. Complete endoscopic resection of the tumour is the first diagnostic and therapeutic step in T1G3 management. A second resection should be done at 1 month to avoid residual tumour and misdiagnosis of a muscle infiltrative cancer. As a result of treatment by instillations of Calmette and Guerin bacillus following endoscopic resection, a 5-year survival rate of 80% has been reported, with 50 to 60% of bladder preservation. BCG is the only conservative treatment that has proven effectiveness on both tumour recurrence and progression. Long term protocols seem to give the best results. Endovesical chemotherapy is not commonly used as its impact on progression has not been demonstrated. Radical cystectomy can be chosen as first line treatment in patients with particularly aggressive tumours. Long term and close surveillance should be achieved in every patient. PMID- 16709008 TI - [Screening for prostate cancer: arguments "against"]. AB - Prostate cancer screening is controversial since PSA assay has been made available. Screening supporters consider that early diagnosis allows better and less aggressive treatment. These arguments lie on longitudinal cohort studies without controls. Randomized studies are required to assess the correlation between screening and mortality lowering. Two studies are being performed and their results will be available within three or four years. Consequently, the validity of screening is unknown. Nevertheless, the analysis of various parameters demonstrates that the reduction of cancer mortality related to screening is low. According to these data and as recommended by health institutions prostate cancer screening is not required. At the present time, detection asked by the patient himself remains the good attitude between negligence and excessive attitude. PMID- 16709009 TI - [Screening for prostate cancer: synthesis]. AB - Screening for prostate cancer (PCa) remains a matter of debate. Although the results of two ongoing randomised trials will be available in 2008, mass screening of PCa is already effective in the United States where PCa-related mortality has very significantly decreased. However, such screening is strongly suspected to induce "over-diagnosis". This results in the treatment of insignificant PCa, but in a significant complication rate. This situation finds an explanation in the low specificity of PSA, the screening tool for aggressive PCa. ALL of the debate is based on a misunderstanding between the pros and the cons of screening. On one hand, if diagnosis is considered a search for knowledge, then advocate an "over-diagnosis" would mean advocate an "over knowledge", which is nonsense. On the other hand, if treatment is considered a search for action on the natural history of a disease, then advocate an "over treatment" would mean advocate an "over-action", which is meaningful. The problem induced by screening is not to search for a disease, but what we do with it when we find it! Before recommending mass screening for PCa, we have to re-educate our patients in their understanding of the disease and re-educate ourselves in our treatment decision process. Waiting for a governmental decision about PCa screening policy, one should actually consider individual screening, which allows for personal education of patients. PMID- 16709010 TI - [Active follow-up of prostate cancers]. AB - Today, earlier diagnosis of prostate cancer has allowed including, beside standard curative treatments, active follow-up of the disease. Such therapeutic management is based on a better knowledge of the natural history of prostate cancer and on possible detection of cases with slow progression. However, all criteria of selection of tumours that are likely to benefit from such surveillance are not always reliable and in some cases invasive treatment should replace surveillance. Analysis of active follow-up series shows that patients who benefit from such treatment have an unchanged prognosis. Active surveillance of prostate cancers may be considered a valuable option provided very strict limits are established regarding patients selection and follow-up. PMID- 16709011 TI - [Scrotal trauma: management strategy]. AB - Scrotal traumas are rare. Most are blunt traumas caused by a direct blow on the scrotum. The testicle is projected against the pubic arch. Early surgical investigation has considerably improved the prognosis of testicular trauma, and reduced orchidectomy rate. ULtrasonography has also improved the management of scrotal trauma. But there is a controversy about accuracy of ultrasonography in predicting presence or absence of testicular disruption. ULtrasonography should not challenge the dogma regarding systematic surgical investigation of hematocele and enlarged scrotum. Long term outcomes (testicular atrophy, infertility) may be more frequent as previously thought and should be detected. PMID- 16709012 TI - [Balanitis: diagnosis and treatment]. AB - Balanitis refers to a variety of unrelated conditions. It results from infective, irritative, allergic, traumatic, or inflammatory causes; pre-malignant lesions have been also identified. All these causes are successively reviewed regarding their positive diagnosis and their treatment. Normal aspects of the balanopreputial area will be explained because they may cause some anxiety in certain patients. The difference between the circumcised and uncircumcised penile skin that cause differences in the incidence and appearance of dermatoses of the glans and corona will be studied. Pre-malignant lesions of the balanopreputial area, although not frequent, represent a difficult diagnosis and therapeutic challenge. The major problem is the earliness of the diagnosis that may avoid the occurrence of squamous cell carcinoma. Moreover, lichen sclerosus may develop into squamous cell carcinoma and therefore a lifelong follow-up must be implemented. Thus, any fixed, chronic or suspicious Lesion must be rapidly assessed by a biopsy. PMID- 16709013 TI - [How to operate kidney puncture in percutaneous nephrolithotomy?]. AB - Obtaining a precise percutaneous calyceal puncture gave way to the development of percutaneous nephrolithotomy, one of the first micro-invasive techniques described in urology. Both radiologist and urologist can perform puncture, sometimes in a collaborative effort. However, being followed by a true surgical procedure, it should be done in the O.R; perfect knowledge of the procedure is mandatory for every urologist. Standard guidance uses a fluoroscopic C-arm device, only able to guide the needle precisely towards the apex of the chosen calyx. Moving the C-arm with cephalad tilting will provide 3-D imaging. Ultrasound guidance is an alternative, but might be difficult with non dilated upper tract. CT guidance and retrograde puncture are rarely used. The access is to be adapted according to the patient (adult or child), type of stone (single or multiple access), or kidney position (eutopic or ectopic). Direct ad stable puncture entering the apex of the chosen calyx is a pre-requisite for easy and efficient subsequent nephrolithotomy. PMID- 16709014 TI - Trends in brain cancer incidence and survival in the United States: Surveillance, Epidemiology, and End Results Program, 1973 to 2001. AB - OBJECT: An increasing incidence of brain cancer has been reported for the last three decades. In this study of brain cancer incidence and patient survival in the US, the authors attempt to update information on trends by examining data provided by the Surveillance, Epidemiology, and End Results (SEER) Program. METHODS: Population-based data from the SEER Program were used to calculate the incidence of and survival rates for people with brain cancer. The approximate Poisson method was used to calculate relative risks for brain cancer and to determine a 95% confidence interval. Annual age-standardized incidence rates were calculated, and time-trend analysis was conducted using joinpoint regression analysis. The relative risks of brain cancer were 1.48 for men compared with women, 3.18 for elderly persons compared with young adults, 1.86 for Caucasian patients compared with African-American patients, and 1.35 for those in metropolitan counties compared with those in nonmetropolitan counties. The incidence of brain cancer increased until 1987, when the annual percentage of change reversed direction, decreasing from 1.68 to 20.44%. The elderly experienced an increase until 1985, but their rates were stable thereafter. Rising trends were noticed for glioblastoma multiforme (GBM), oligodendroglioma, anaplastic astrocytoma, medulloblastoma, and mixed glioma, and falling trends were observed for astrocytoma not otherwise specified and malignant glioma. The survival rate for patients with GBM has not shown improvement in the last two decades. CONCLUSIONS: Increased risk of brain cancer is associated with being male, Caucasian, elderly, and residing in a metropolitan county. The incidence rate of brain cancer in the US is gradually declining, but the rising trend of GBM combined with its poor survival rate is disconcerting and needs further exploration. PMID- 16709015 TI - Convection-enhanced delivery: targeted toxin treatment of malignant glioma. AB - Historically, malignant gliomas are perhaps the most difficult intracranial neoplasms to treat. Surgery, radiation therapy, and traditional chemotherapy have not been able to significantly alter the course of this disease. By definition, these tumors are located in the protected space of the cranial vault, where the blood-brain barrier prevents most therapies from gaining access. Because of the difficulty in treating this disease, new, innovative treatments and alternative delivery techniques for those therapies are needed. Targeted toxins are fusion proteins that represent a novel medical treatment for these cancers that is under development. However, the efficacy of these agents is dependent on the method of delivery to the tumor. The administration of targeted toxins requires image guided placement of catheters, either within the tumor or into the adjacent infiltrated brain, and positive pressure infusion. The term that has been applied to this microinfusion technique is convection-enhanced delivery (CED). This infusion method was first attempted via direct intratumoral infusion in nude mouse flank tumor models of human malignant glioma. After significant development of this delivery technique in animal models, the successful demonstration of in vivo efficacy of targeted toxins in Phase I and II clinical trials was reported. Currently, ongoing targeted toxin trials are being conducted at academic health centers to define the best clinical practice for CED. This work involves refining the details of delivery such as infusion rate, duration of treatment, and drug dosing. The early results of CED of targeted toxins supports their continued investigation, as few other treatment modalities have produced durable results in the fight against gliomas. PMID- 16709016 TI - The IL-4 and IL-13 pseudomonas exotoxins: new hope for brain tumor therapy. AB - Targeting cell surface receptors with cytotoxins or immunotoxins provides a unique opportunity for brain tumor therapy. The authors have discovered that receptors for two cytokines, interleukin (IL)-4 and IL-13, are overexpressed on tumor biopsy samples and on cell lines derived from a variety of human tumors, including brain tumors. These investigators have demonstrated that the structure of these cytokine receptors on tumor cells is different from that found on normal immune cells. In human solid tumor cells, IL-4 binds to two chains (IL-4Ra and IL 13Ra1), whereas IL- 13 binds to three chains in many solid tumor cells, including glioma cells (to IL-4Ra, IL-13Ra1, and IL-13Ra2). To target IL-4Rs and IL-13Rs, the authors generated two recombinant fusion cytotoxins composed of IL-4 or IL-13 and a mutated form of pseudomonas exotoxin (PE), which for simplicity are called IL4-PE and IL13-PE in this paper. These chimeric cytotoxins are highly toxic in vitro to human tumor cell lines and primary cell cultures, including glioma cells, and in vivo to animal models of human tumors, including gliomas. In contrast, normal cells, including immune, endothelial, and brain cells, are spared from their cytotoxic effects. Based on numerous preclinical studies, IL13 PE (also known as IL13-PE38QQR or cintredekin besudotox) has been tested in four Phase I/II clinical trials. The agent IL13-PE was administered intracranially by using convection-enhanced delivery (CED). The drug was delivered through catheters placed either directly into the tumor bed or in the peritumoral region after resection of the lesion. The CED of IL13-PE was fairly well tolerated, with a reasonable benefit/risk profile for treatment of patients with glioma. Based on Phase I/II clinical trials, the Phase III Randomized Evaluation of CED of IL13-PE Compared to Gliadel Wafer with Survival Endpoint Trial (also known as the PRECISE Trial) in patients with initial recurrence of glioblastoma multiforme has recently been completed. Patients are being monitored for safety of the agents, duration of overall survival, and quality of life. PMID- 16709017 TI - Convection-enhanced delivery of therapeutics for brain disease, and its optimization. AB - Convection-enhanced delivery (CED) is the continuous injection under positive pressure of a fluid containing a therapeutic agent. This technique was proposed and introduced by researchers from the US National Institutes of Health (NIH) by the early 1990s to deliver drugs that would otherwise not cross the blood-brain barrier into the parenchyma and that would be too large to diffuse effectively over the required distances were they simply deposited into the tissue. Despite the many years that have elapsed, this technique remains experimental because of both the absence of approved drugs for intraparenchymal delivery and the difficulty of guaranteed delivery to delineated regions of the brain. During the first decade after the NIH researchers founded this analytical model of drug distribution, the results of several computer simulations that had been conducted according to more realistic assumptions were also published, revealing encouraging results. In the late 1990s, one of the authors of the present paper proposed the development of a computer model that would predict the distribution specific to a particular patient (brain) based on obtainable data from radiological images. Several key developments in imaging technology and, in particular, the relationships between image-obtained quantities and other parameters that enter models of the CED process have been required to implement this model. Note that delivery devices need further development. In the present paper we review key features of CED as well as modeling of the procedure and indulge in informed speculation on optimizing the direct delivery of therapeutic agents into brain tissue. PMID- 16709018 TI - Convection-enhanced delivery of immunotoxins and radioisotopes for treatment of malignant gliomas. AB - The treatment of malignant gliomas has advanced significantly in the past 15 years. The simultaneous development of new targeting agents and techniques to deliver these high-molecular-weight compounds has led to improved efficacy and promising results in Phase III trials. Convection-enhanced delivery (CED) of macromolecules has emerged as the leading delivery technique for the treatment of malignant gliomas. A summary of the basic principles of CED and a review of the current human trials of protein targeting agents are provided. PMID- 16709019 TI - Comparison of intratumoral bolus injection and convection-enhanced delivery of radiolabeled antitenascin monoclonal antibodies. AB - OBJECTIVES: Convection-enhanced delivery (CED) is a novel technique used to deliver agents to the brain parenchyma for treatment of neoplastic, infectious, and degenerative conditions. The purpose of this study was to determine if CED would provide a larger volume of distribution (Vd) of a radiolabeled monoclonal antibody (mAb) than a bolus injection. METHODS: Patients harboring a recurrent glioblastoma multiforme that reacted with the antitenascin mAb 81C6 during immunohistochemical analysis were randomized to receive an intratumoral injection of the human-murine chimeric mAb Ch81C6, which had been labeled with the 123I tracer. The mAb was administered by either a bolus injection or CED via a stereotactically placed catheter; between 48 and 72 hours later the mAb was again administered using the other technique. Injections of escalating doses of a 131I labeled therapeutic mAb were then delivered using the technique shown to produce the largest Vd by single-photon emission computerized tomography. CONCLUSIONS: Convection-enhanced delivery has enormous potential for administering drugs to sites within the central nervous system. For the relatively small volumes injected in this study, however, CED did not provide a significant increase in the Vd when compared with the bolus injection. Nevertheless, a clear cross-over effect was seen, which was probably related to the temporal proximity of the two infusions. PMID- 16709020 TI - Safety of intraparenchymal convection-enhanced delivery of cintredekin besudotox in early-phase studies. AB - OBJECT: Convection-enhanced delivery (CED) is an increasingly used novel local/regional delivery method targeted directly to tissue. It relies on a continuous pressure gradient for distribution of therapeutic agents into the interstitial space, with administration of the infusate over a few days. Cintredekin besudotox (also known as IL13- PE38QQR) is a recombinant chimeric cytotoxin consisting of interleukin-13 and a truncated exotoxin produced by the Pseudomonas aeruginosa bacterium, which targets malignant glioma cells. METHODS: Cintredekin besudotox was administered via intraparenchymal CED after resection of supratentorial recurrent malignant glioma. The safety and toxicity profile was reviewed for 53 patients in whom infusion catheters had been placed; 51 of them received CED of the study drug. Adverse events were categorized based on time of onset in relation to CED, and the causal relationship with catheter placement or delivery of cintredekin besudotox. Catheters were placed in 53 patients, although only 51 of them received cintredekin besudotox. Most adverse events related to catheter placement or the study drug originated from the central nervous system. Three symptomatic windows were defined: the first one was between surgical procedure and CED; the second was during CED and up to 1 week after its completion; and the third window was 2 to 10 weeks after treatment. Those windows generally reflected adverse events related to surgical procedures, mass effect from infusate, and drug effect on tumor-infiltrated and normal brain parenchyma, respectively. CONCLUSIONS: The symptomatic windows identified in this study apply to any CED clinical trials, particularly those in which chimeric cytotoxins are used, and will help to determine the most likely underlying pathophysiological process causing symptoms. This information, in turn, will help to prevent adverse events or minimize their severity. Those events also have implications for dose escalation and outcome measures. PMID- 16709021 TI - Image-guided resection of high-grade glioma: patient selection factors and outcome. AB - OBJECT: In patients with glioma, image-guided surgery helps to define the radiographic limits of the tumor to maximize safety and the extent of resection while minimizing damage to eloquent brain tissue. The authors hypothesize that image-guided resection (IGR) techniques are associated with improved outcomes in patients with malignant glioma. METHODS: Data recorded in 486 patients enrolled in the Glioma Outcomes Project were analyzed in this study. Demographic data and outcomes in patients who underwent IGR were compared with those in patients who underwent resection without IGR. Univariate analysis performed with chi-square testing was used to compare patient presentation, tumor characteristics, and death rates. Multivariate logistic regression was used to predict various outcome parameters. Patients who underwent IGR were younger and had smaller, lower-grade tumors than those in whom IGR was not performed. They were more likely to present with seizure and normal consciousness. Unexpectedly, gross-total resection was performed in significantly fewer patients with IGR than in individuals without IGR. Patients with IGR were more likely to be discharged home with the ability to live independently, and they had a shorter duration of hospital stay than patients without IGR. Survival was significantly longer in patients who underwent IGR, but multivariate analysis showed that glioblastoma multiforme (GBM) and age accounted for these observations. CONCLUSIONS: Selection bias occurs regarding patients who receive IGR; these biases include younger age, presentation with seizure and normal level of consciousness, tumor diameter less than 4 cm, and non GBM on histopathological studies. Outcome appears to be improved in patients who undergo IGRs of high-grade gliomas. It is unclear if these improved outcomes are due to the selection of a more favorable patient population or to the IGR techniques themselves. It is likely that the full potential of image guidance in glioma surgery will not be realized until it is applied to a wider range of patients. PMID- 16709022 TI - Gamma knife surgery for glioblastoma multiforme. AB - Despite the implementation of increasingly aggressive surgery, chemotherapy, and fractionated radiotherapy for the treatment of glioblastoma multiforme (GBM), most therapeutic regimens have resulted in only modest improvements in patient survival. Gamma knife surgery (GKS) has become an indispensable tool in the primary and adjuvant management of many intracranial pathologies, including meningiomas, pituitary tumors, and arteriovenous malformations. Although it would seem that radiosurgical techniques, which produce steep radiation dose fall-off around the target, would not be well suited to treat these infiltrative lesions, a limited number of institutional series suggest that GKS might provide a survival benefit when used as part of the comprehensive management of GBM. This may largely be attributed to the observation that tumors typically recur within a 2-cm margin of the tumor resection cavity. Despite these encouraging results, enthusiasm for radiosurgery as a primary treatment for GBM is significantly tempered by the failure of the only randomized trial that has been conducted to yield any benefit for patients with GBM who were treated with radiosurgery. In this paper, the authors review the pathophysiological mechanisms of GKS and its applications for GBM management. PMID- 16709023 TI - Gene therapy for glioblastoma. AB - Established treatments such as surgery, radiation, and chemotherapy have only minimally altered the median survival time of patients with glioblastoma multiforme, the most common malignant brain tumor. These failures reflect the highly invasive nature of the disease, as well as the fact that few cells are actively dividing at any given time. As a result, therapies need to act in areas of the brain that are spatially separated from the site of tumor origin and over extended periods of time temporally separated from their introduction. Over the past decade, laboratory studies and early clinical trials have raised the hope that these therapeutic requirements may be fulfilled by gene therapy in which nonreplicating transgene-bearing viruses, oncolytic viruses, or migratory stem cells are used to deliver tumoricidal transgenes. The authors review the principles behind these approaches and their initial results. PMID- 16709024 TI - Oncolytic adenoviral therapy for glioblastoma multiforme. AB - Adenoviruses historically have been one of the main vectors used in human gene therapy. To date, the majority of brain tumor trials of these vectors have used replication-defective viruses. The relative lack of success obtained with replication-defective vectors has prompted a search for new and improved therapies. In this context, oncolytic (conditionally replicative) adenoviruses, which selectively bind and replicate only in tumor cells, have gained increasing importance. These adenoviruses, once they are rendered conditionally replicative by transductional and transcriptional modifications, offer significant promise for patients with malignant glioma. In this review, the authors discuss the genetic approaches to adenoviral modification and their applications in the field of neurooncology. PMID- 16709025 TI - Treatment options for glioblastoma. AB - Following the seminal trial conducted by the European Organisation for Research and Treatment of Cancer (EORTC) and the National Cancer Institute of Canada (NCIC), concurrent temozolomide and radiotherapy has become the new standard of care for patients with newly diagnosed glioblastoma multiforme (GBM). Investigation of emerging therapies (which are now used as salvage therapy) such as small-molecule inhibitors (for example, epidermal growth factor receptor inhibitors) and convection-enhanced delivery (CED) of targeted toxins (for example, interleukin-13/pseudomonas exotoxin) is likely to build on the EORTC/NCIC treatment platform and will, it is hoped, improve survival rates in patients with GBM. The majority of adjuvant Phase I and II trials being conducted by the brain tumor consortia are based on the EORTC/NCIC treatment platform and have added a targeted therapy in an effort to find a promising synergistic treatment. Furthermore, researchers in the consortia are continuing to explore treatments for recurrent GBM, not otherwise eligible for local therapies, such as CED. The treatments under study include novel cytotoxic chemotherapy as well as small-molecule inhibitors; these are being assessed in a variety of Phase I or II trials. PMID- 16709026 TI - Enhancement of glioblastoma cell killing by combination treatment with temozolomide and tamoxifen or hypericin. AB - OBJECT: The chemotherapeutic agent temozolomide has demonstrated antitumor activity in patients with recurrent malignant glioma. Because responses are not enduring and recurrence is nearly universal, further improvements are urgently needed. METHODS: In an effort to increase the clinical activity of temozolomide, the authors investigated whether its antitumor activity could be enhanced by adding tamoxifen or hypericin, two drugs that are known to inhibit the activity of protein kinase C. Human glioblastoma multiforme cell lines A172 and LA567 were treated with combinations of temozolomide and tamoxifen or hypericin in vitro, and cell survival was analyzed using various methods. Tamoxifen and hypericin were able to greatly increase the growth-inhibitory and apoptosis-stimulatory potency of temozolomide via the downregulation of critical cell cycle-regulatory and prosurvival components. Furthermore, with the use of an in vivo xenograft mouse model, the authors demonstrated that hypericin was able to enhance the antiglioma effects of temozolomide in the in vivo setting as well. CONCLUSIONS: Taken together, analysis of the results indicated that combination therapy involving temozolomide and tamoxifen or hypericin potently inhibited tumor growth by inducing apoptosis and provided an effective means of treating malignant glioma. PMID- 16709027 TI - Antitumor properties of dimethyl-celecoxib, a derivative of celecoxib that does not inhibit cyclooxygenase-2: implications for glioma therapy. AB - Celecoxib (Celebrex) appears to be unique among the class of selective COX-2 inhibitors (coxibs), because this particular compound exerts a second function that is independent of its celebrated ability to inhibit COX-2. This second function is the potential to inhibit cell proliferation and stimulate apoptotic cell death at much lower concentrations than any other coxibs. Intriguingly, these two functions are mediated by different moieties of the celecoxib molecule and can be separated. The author, as well as others, have generated and investigated analogs of celecoxib that retain only one of these two functions. One derivative, 2,5-dimethyl-celecoxib (DMC), which retains the antiproliferative and apoptosis-inducing function, but completely lacks the COX-2 inhibitory activity, is able to mimic faithfully all of the numerous antitumor effects of celecoxib that have been investigated so far, including reduction of neovascularization and inhibition of experimental tumor growth in various in vivo tumor models. In view of the controversy that has recently arisen regarding the life-threatening side effects of this class of coxibs, it may be worthwhile to pursue further the potential benefits of drugs such as DMC for anticancer therapy. Because DMC is not a coxib yet potently maintains celecoxib's antitumor potential, one may be inclined to speculate that this novel compound could potentially be advantageous in the management of COX-2-independent cancers. In this summary, the implications of recent findings with DMC will be presented and discussed. PMID- 16709028 TI - Characteristics of tumor-associated endothelial cells derived from glioblastoma multiforme. AB - Glioblastomas multiforme (GBMs) are highly vascular brain tumors characterized by abnormal vessel structures in vivo. This finding supports the theory that glioma associated endothelial cells (ECs) have intrinsically different properties from ECs in normal human brain. Therefore, identification of the functional and phenotypic characteristics of tumor-associated ECs is essential for designing a rational antiangiogenic therapy. The GBM-associated ECs have a large, flat, and veil-like appearance, in contrast to normal ones, which are small and plump. Although the tumor ECs have the typical markers, they proliferate more slowly than these cell types in normal brain. The GBM-associated ECs are resistant to cytotoxic drugs, and they undergo less apoptosis than control cells. Also, GBM associated ECs migrate faster than controls and constitutively produce high levels of growth factors such as endothelin-1, interleukin-8, and vascular endothelial growth factor. An understanding of these unique characteristics of glioma-associated ECs is important for the development of novel antiangiogenic agents that specifically target tumor-associated ECs in gliomas. PMID- 16709030 TI - Hypoxia in the tumorigenesis of gliomas and as a potential target for therapeutic measures. AB - In this article, the author provides a brief description of the role of hypoxia in the tumorigenesis of gliomas and suggests potential ways of exploiting this role to design treatment modalities. Tumor hypoxia predicts the likelihood of metastases, tumor recurrence, resistance to chemotherapy and radiation therapy, invasive potential, and decreased patient survival for many human malignancies. Various methods of measurement of tumor hypoxia are discussed, including direct measurement and imaging methods. The role of hypoxia-responsive molecules, especially hypoxia-inducible factor-1 (HIF-1), in glioma tumorigenesis is explored. Treatment modalities regulated by hypoxia are proposed and some potential strategies reviewed. The progression of a low-grade astrocytoma to a glioblastoma multiforme may be mediated by hypoxia-induced phenotypic changes and subsequent clonal selection of cells that overexpress hypoxia-responsive molecules, such as HIF-1. In this model, intratumoral hypoxia causes genetic changes that produce a microenvironment that selects for cells of a more aggressive phenotype. PMID- 16709029 TI - Efficacy of antiangiogenic targeted toxins against glioblastoma multiforme. AB - OBJECT: Because the prognosis for patients with glioblastoma multiforme (GBM) remains poor, investigators have focused on developing new and more effective treatment modalities. Targeted toxins represent a new class of compounds composed of a potent protein toxin and a carrier ligand that will recognize cell surface antigens located on target tissue. A recombinant fusion protein was created that contains the translocation and catalytic portions of diphtheria toxin that are responsible for cell entry and killing, respectively, fused to the noninternalizing aminoterminal fragment portion of human plasminogen activator. This diptheria toxin-uPA fusion protein (DTAT) has the advantage over other fusion proteins of targeting malignant glioma cells and the endothelial cells of the neovasculature that express the urokinase-type plasminogen activator receptor (uPAR). Another protein, DTAT13, was synthesized to target uPAR on the neovasculature and the uPAR and interleukin-13 receptor-expressing GBM cells. The authors describe the in vitro and in vivo efficacy of DTAT and DTAT13 against GBM. METHODS: The in vitro cytotoxicity of DTAT and DTAT13 was measured using cell proliferation assays. In vivo studies were performed in which DTAT, DTAT13, or a control protein was injected directly into GBM flank tumors in athymic nude mice. Tumor volume was assessed over time and analyzed using the Student t-test. The systemic organ effects of DTAT and DTAT13 were examined functionally and histologically in tumor-free C57BL/6 mice. In vitro, DTAT and DTAT13 were found to be highly potent and selective against U118MG, U87MG, and U373MG GBM cell lines and human umbilical vein endothelial cells. In vivo, DTAT and DTAT13 both caused a statistically significant (p < 0.05) regression of U87MG GBM flank tumors when administered every other day at 10 mg/day for five doses. No tumor regression was seen in control animals. Both DTAT and DTAT13 had little effect on histological findings in the liver, kidney, spleen, and lungs. Serum analysis did not demonstrate an effect on blood urea nitrogen levels, but liver alanine aminotransferase levels rose to statistically significant (p = 0.046) but not life-threatening levels. Also, DTAT13 was less toxic than DTAT in studies of mortality rates. CONCLUSIONS: Both DTAT and DTAT13 might have potential for clinical application against GBM because of their ability to target both the tumor cells and neovasculature simultaneously with an absence of serious systemic side effects. The discovery that DTAT13 was less toxic than DTAT indicated that the bispecific fusion protein might target a broader subset of antigenetically diverse patients with tumors while reducing the systemic exposure to toxin that would be necessary if two agents were administered separately. PMID- 16709032 TI - Fibroblast growth factor-2-retargeted adenoviral vector for selective transduction of primary glioblastoma multiforme endothelial cells. AB - OBJECT: Adenovirus transduction in gene therapy is dependent on the expression of the coxsackie virus-adenovirus receptor (CAR) for initial binding and on the integrin receptors (avb3, avb5) for viral internalization. Low and variable expression of CAR may be responsible for the low transduction rates seen with native adenoviral vectors. The goal of this study was to demonstrate increased transduction efficiency by retargeting the adenovirus with a fibroblast growth factor (FGF) ligand, FGF-2. METHODS: The retargeted adenoviruses were used to transduce human glioblastoma multiforme (GBM)-derived ECs (tumor-associated brain endothelial cells [TuBECs]), in which there is minimal CAR expression but a high expression of FGF receptor (FGFR). The results demonstrate that the transduction efficiency of TuBECs can reach as high as 80% when one uses an FGF2-conjugated adenovirus containing green fluorescent protein (FGF2-AdGFP) yet be only 5% when one uses the native adenovirus (AdGFP). The TuBECs were transduced with either a native adenovirus (AdHSV-TK) or a retargeted adenovirus (FGF2-AdHSV-TK), both of which carry the suicide herpes simplex virus-thymidine kinase (HSV-TK) gene. Administered as a cytotoxic prodrug, ganciclovir induced a significant decline in the proliferation rate and increased apoptosis in TuBECs treated with the retargeted adenovirus, compared with its effect on TuBECs treated with the native adenovirus. Increased transduction efficiency was determined by performing GFP based flow cytometry, and the expression of the TK protein by the retargeted adenovirus was assessed by performing an immunohistochemical analysis focused on HSV-TK. The mechanism of cytotoxicity was determined to be apoptosis by performing a terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling assay. CONCLUSIONS: Fibroblast growth factor-2 retargeted adenoviral vectors may be used to increase the transduction of GBM derived endothelial cells, enabling a new and efficient antiangiogenesis strategy for the treatment of malignant gliomas. PMID- 16709031 TI - Use of replication-competent retroviral vectors in an immunocompetent intracranial glioma model. AB - OBJECT: The authors had previously reported on a replication-competent retrovirus (RCR) that has been demonstrated to be stable, capable of effective transduction, and able to prolong survival in an intracranial tumor model in nude mice. The purpose of this study was further investigation of this gene therapy option. METHODS: The transduction efficiency of RCR in RG2, an immunocompetent intracranial tumor model, was tested in Fischer 344 rats. The immune response to the RCR vector was expressed by the quantification of CD4, CD8, and CD11/b in tumors. The pharmaceutical efficacy of the suicide gene CD in converting prodrug 5-fluorocytosine (5-FC) to 5-fluorouracil (5-FU) was measured using fluorine-19 nuclear magnetic resonance (19F-NMR) spectroscopy. Animal survival data were plotted on Kaplan-Meier survival curves. Finally, the biodistribution of RCR was determined using quantitative real-time polymerase chain reaction (RT-PCR) for the detection of retroviral env gene. There was no evidence of viral transduction in normal brain cells. Neither severe inflammation nor immunoreaction occurred after intracranial injection of RCR-green fluorescent protein compared with phosphate-buffered saline (PBS). The 19F-NMR spectroscopy studies demonstrated that RCR-CD was able to convert 5-FC to 5-FU effectively in vitro. The infection of RG2 brain tumors with RCR-CD and their subsequent treatment with 5-FC significantly prolonged survival compared with that in animals with RG2 transduced tumors treated with PBS. In contrast to the nude mouse model, evidence of virus dissemination to the systemic organs after intracranial injection was not detected using RT-PCR. CONCLUSIONS: The RCR-mediated suicide gene therapy described in this paper effectively transduced malignant gliomas in an immunocompetent in vivo rodent model, prolonging survival, without evidence of severe intracranial inflammation, and without local transduction of normal brain cells or systemic organs. PMID- 16709033 TI - Brain tumor stem cells: new targets for clinical treatments? AB - The observation of similarities between the self-renewal mechanisms of stem cells and cancer cells has led to the new concept of the cancer stem cell. In cases of leukemia, multiple myeloma, and breast cancer, cells with a high selfrenewal potential have been identified. Furthermore, investigators have shown these cells' ability to drive the formation and growth of the tumor. Brain tumors have also been reported to possess a subpopulation of cancer stemlike cells that have the ability to proliferate, self-renew, and be multipotent. When grafted into mice, these cells are also able to generate a tumor that recapitulates that of the patient from whom the cells were derived. The identification and characterization of this new category of cells call for new therapies capable of selectively targeting and killing these multifaceted cells. PMID- 16709034 TI - Management of malignant glioma: steady progress with multimodal approaches. AB - Despite recent successes in the treatment of cancer with multidisciplinary multimodal treatment approaches, the duration of survival for patients with malignant glioma remains limited. Malignant gliomas represent a class of infiltrative, aggressive neoplasms that are generally resistant to combination therapies. The basic approach to treatment has involved a combination of surgery and radiotherapy. The use of chemotherapy has been met with skepticism because of its limited efficacy and the significant side effects demonstrated in clinical trials. Nevertheless, based on findings in randomized trials of new agents, it has been suggested that further evaluation of the role of chemotherapy is warranted. Temozolomide and Gliadel (carmustine wafers) are generally well tolerated due to their limited systemic toxicity. These agents appear particularly well suited for incorporation into multimodal treatment strategies. Proposed investigations and ongoing clinical trials will be conducted to assess the use of these agents in novel combination therapies. Future treatment strategies may include a wide variety of biological response modifiers, but will need to continue to address local control with surgery, radiation, and adjuvant chemotherapy. PMID- 16709035 TI - Standard treatment and experimental targeted drug therapy for recurrent glioblastoma multiforme. AB - Glioblastoma multiforme (GBM) tumors almost invariably recur despite initial treatments. Correct diagnosis using a variety of imaging techniques and the involvement of a multidisciplinary tumor board are critical for evaluating each stage of a patient's progression and determining optimal management. Standard therapies for recurrence generally include repeated resection, radiation therapy, chemotherapy, and supportive care; however, salvage therapy must be highly individualized, and not all patients are eligible for every type of standard therapy. Factors such as the size and location of the tumor, previous treatment, and general health of the patient must be taken into consideration. Although standard therapies can prolong a patient's duration of survival, the median survival time for patients with recurrent GBM is usually less than 1 year. Experimental targeted drug therapies have been developed to inhibit aberrant cell signaling pathways involved in tumorigenesis, and enrolling patients in clinical trials using these therapies is another option for treatment of recurrent GBM. The use of these novel therapies is often confined to large research institutions, but the severe limitations of standard treatment options make it important to highlight the potential of experimental therapies. In this paper the authors outline standard therapies and review the emerging role of targeted drug therapy in the treatment of recurrent GBM. PMID- 16709036 TI - Recurrent glioblastoma multiforme: a review of natural history and management options. AB - Glioblastoma multiforme (GBM) is one of the most aggressive primary brain tumors, with a grim prognosis despite maximal treatment. Advancements in the past decades have not significantly increased the overall survival of patients with this disease. The recurrence of GBM is inevitable, its management often unclear and case dependent. In this report, the authors summarize the current literature regarding the natural history, surveillance algorithms, and treatment options of recurrent GBM. Furthermore, they provide brief discussions regarding current novel efforts in basic and clinical research. They conclude that although recurrent GBM remains a fatal disease, the literature suggests that a subset of patients may benefit from maximal treatment efforts. Nevertheless, further research effort in all aspects of GBM diagnosis and treatment remains essential to improve the overall prognosis of this disease. PMID- 16709037 TI - New trends in the medical management of glioblastoma multiforme: the role of temozolomide chemotherapy. AB - Standard care for newly diagnosed glioblastoma multiforme (GBM) previously consisted of resection to the greatest extent feasible, followed by radiotherapy. The role of chemotherapy was controversial and its efficacy was marginal at best. Five years ago temozolomide (TMZ) was approved specifically for the treatment of recurrent malignant glioma. The role of TMZ chemotherapy administered alone or as an adjuvant therapy for newly diagnosed GBM has been evaluated in a large randomized trial whose results suggested a significant prolongation of survival following treatment. Findings of correlative molecular studies have indicated that methylguanine methyltransferase promoter methylation may be used as a predictive factor in selecting patients most likely to benefit from such treatment. In this short review the authors summarize the current role of TMZ chemotherapy in the management of GBM, with an emphasis on approved indications and practical aspects. PMID- 16709038 TI - Autophagy, the Trojan horse to combat glioblastomas. AB - Malignant gliomas, among which glioblastomas constitute the largest group, are characterized by a dramatically diffuse infiltration into the brain parenchyma with, as a consequence, the fact that no patient with glioblastoma multiforme (GBM) has been cured to date. Migrating GBM cells are resistant to apoptosis (Type I programmed cell death), and thus to radiotherapy and conventional chemotherapy, because of the constitutive activation of several intracellular signaling pathways, of which the most important identified to date are the pathways controlled by phosphatidylinositol 3-kinase, Akt, and the mammalian target of rapamycin (mTOR). Migrating GBM cells seem to be less prone to resist autophagy (Type II programmed cell death), and disruption of the pathway controlled by mTOR induces marked autophagic processes in GBM cells. Temozolomide is the most efficacious cytotoxic drug employed today to combat glioblastoma, and this drug exerts its cytotoxic activity through proautophagic processes. Thus, autophagy represents a kind of Trojan horse that can be used to bypass, at least partly, the dramatic resistance of glioblastoma to radiotherapy and proapoptotic related chemotherapy. PMID- 16709039 TI - Cellular signaling molecules as therapeutic targets in glioblastoma multiforme. AB - Despite recent advances in operative techniques, chemotherapy, and radiotherapy, the prognosis in patients with glioblastoma multiforme (GBM) remains poor; the majority die within a year of diagnosis. Although often effective at reducing mass effect and tumor burden, surgical debulking and cytotoxic therapies have never demonstrated an unequivocally significant benefit in treating patients with GBM. This shortcoming has led to the development of molecules that target specific steps in the transduction pathways of high-grade glioma cells. In this article the authors review various cellular and extracellular signaling pathways that may prove promising in the treatment of patients with malignant glioma. PMID- 16709041 TI - Studies on the reactive species in fluoride-mediated carbon-carbon bond-forming reactions: carbanion formation by desilylation with fluoride and enolates. AB - The reactive species in fluoride-mediated carbon-carbon bond-forming reactions was investigated. The regio- and diastereoselectivities of silanes reacting with cyclohexenone in the presence of a catalytic amount of fluoride was compared to the reactivity of analogous solvent-separated lithium ion pairs. Closely analogous behavior was observed, showing that carbanions and not siliconate complexes are the reactive species in the fluoride-catalyzed reactions. Spectroscopic investigations unambiguously show that phenylthiobenzyl anion will form by reaction of silane with tris(dimethylamino)sulfonium difluorotrimethylsilicate (TASF) or crypt[2.1.1]-solvated lithium enolates. The catalytic cycle runs smoothly with the crypt[2.1.1] complex of alpha (phenylthio)benzyllithium as the initiator and enolate as the carrier of the desilylation reaction. PMID- 16709040 TI - Novel therapies for malignant gliomas: a local affair? AB - Advances in medical and surgical treatments in the last few decades have resulted in quantum leaps in the overall survival of patients with malignant disease outside the central nervous system, whereas survival of patients with malignant gliomas (World Health Organization Grades III and IV) has remained essentially unchanged. Resection and external-beam fractionated radiotherapy remain the pillars of therapy for malignant gliomas and have shown significant beneficial effects on outcome in many clinical studies. On the other hand, numerous human trials with adjuvant agents, most of them administered systemically and causing serious complications and side effects, have not succeeded in achieving a noteworthy additional extension of survival duration, or have done so only with a considerable deterioration in the quality of life of the treated patients. The concept of local invasiveness of gliomas is not new, but only in the last one to two decades has significant attention been focused on the cell biology and molecular genetics of gliomas. Improved understanding of the fundamental features of tumor cells has resulted in the introduction and increasing clinical use of local therapies in which practitioners opt for spatially defined delivery methods and tumor-selective agents specifically designed to be used in the environment of a brain invaded by glioma. In this review, the authors summarize the key findings in some of the most important clinical studies of locally administered treatments for malignant glioma. A few such therapies have emerged in the last decade, and have shown considerable antitumor activity and a favorable profile of local and systemic side effects. These include biodegradable polymers for interstitial chemotherapy, targeted toxins administered by convection-enhanced delivery, and intra- and peritumorally injected genetically modified viruses conferring glioma selective toxicity. In addition, areas of possible improvement of these therapies and essential further developments are outlined. PMID- 16709042 TI - Computational study on the 1,2-rearrangement in beta-(nitroxy)vinyl and beta (acetoxy)vinyl radicals. AB - The 1,2-nitroxyl and 1,2-acetoxyl rearrangement in beta-(nitroxy)vinyl and beta (acetoxy)vinyl radicals 13a and 13b, respectively, has been studied for the gas phase with various ab initio and density functional methods. The energetically most favorable pathway for 13a is calculated to proceed via reversible fragmentation/radical addition through transition state I-19a. In the case of 13b, rearrangement through a five-membered ring transition state III-16b and the fragmentation/radical addition pathway via transition state I-19b are competing processes. Mulliken and natural population analysis reveal a certain degree of charge separation in III-16a/b that may indicate a potential solvent effect on the rearrangement rate. A stepwise group migration through a cyclic radical intermediate V-18a/b or rearrangement through a three-membered ring transition state II-15a/b can be ruled out for both vinyl radicals. A comparison of the results of the calculations with experimental findings provides important insights into the kinetics of "self-terminating radical oxygenations". A significant method dependence on the outcome of the calculations was observed, which revealed the unsuitability of the UHF, MP2, B3LYP, and mPW1PW91 methods for computing these radical rearrangement processes. The results from BHandHLYP/cc pVDZ calculations showed the best agreement with single-point energy calculations performed at the QCISD and CCSD(T) levels of theory. PMID- 16709043 TI - Nitrenes, diradicals, and ylides. Ring expansion and ring opening in 2 quinazolylnitrenes. AB - Tetrazolo[1,5-a]quinazoline (9) is converted to 2-azidoquinazoline (10) on sublimation at 200 degrees C and above, and the azide-tetrazole equilibrium is governed by entropy. 2-Quinazolylnitrenes 11 and 27 and/or their ring expansion products 14 and 29 can undergo type I (ylidic) and type II (diradicaloid) ring opening. Argon matrix photolysis of 9/10 affords 2-quinazolylnitrene (11), which has been characterized by ESR, UV, and IR spectroscopy. A minor amount of a second nitrene, formed by rearrangement or ring opening, is also observed. A diradical (19) is formed rapidly by type II ring opening and characterized by ESR spectroscopy; it decays thermally at 15 K with a half-life of ca. 47 min, in agreement with its calculated facile intersystem crossing (19T --> 19OSS) followed by facile cyclization/rearrangement to 1-cyanoindazole (21) (calculated activation barrier 1-2 kcal/mol) and N-cyanoanthranilonitrile (22). 21and 22 are the isolated end products of photolysis. 21 is also the end product of flash vacuum thermolysis. An excellent linear correlation between the zero-field splitting parameter D (cm(-1)) and the spin density rho on the nitrene N calculated at the B3LYP/EPRIII level is reported (R2 = 0.993 for over 100 nitrenes). Matrix photolysis of 3-phenyltetrazolo[1,5-a]quinazoline (25) affords the benzotriazacycloheptatetraene 29, which can be photochemically interconverted with the type I ring opening product 2-isocyano-alpha-diazo-alpha-phenyltoluene (33) as determined by IR and UV spectroscopy. The corresponding carbene 37, obtained by photolysis of 33, was detected by matrix ESR spectroscopy. PMID- 16709044 TI - Ipso-chlorosulfonylation of calixarenes: a powerful tool for the selective functionalization of the large rim. AB - In our quest for the elaboration of supramolecular models of metallo-enzyme active sites, we became interested in developing new methodologies for the selective functionalization of the large rim of calix[6]arenes. Here, we describe a novel reaction, i.e. the ipso-chlorosulfonylation of calixarene derivatives. The process has been found to be highly efficient, selective and versatile. The regioselectivity is controlled by the nature of the O-substituents at the small rim. Indeed, when O-alkylated by a protonable imidazole group, the aromatic rings are deactivated toward an electrophilic attack and the anisol units can be selectively ipso-chlorosulfonylated under mild conditions (rt). Performing the reaction at a higher temperature allowed the per-chlorosulfonylation to take place. Hence, the synthesis of various sulfonate and sulfonamide derivatives is reported. Finally, a combination of ipso-nitration and chlorosulfonylation allows the per-functionalization of the aromatic units at the large rim in selective alternate positions. Overall, this novel methodology opens new routes to a variety of calixarenes, allowing the tuning of their physical properties without drastically altering their hydrophobic conic cavities. PMID- 16709045 TI - Cyclopropenes in the 1,3-dipolar cycloaddition with carbonyl ylides: experimental and theoretical evidence for the enhancement of sigma-withdrawal in 3-substituted cyclopropenes. AB - The carbonyl ylide dipoles generated by the dirhodium tetra-acetate-catalyzed decomposition of diazocarbonyl precursors 1, 5, and 8 cycloadd to 3-substituted 1,2-diphenylcyclopropenes 3a-e and 3,3-disubstituted cyclopropenes 13, 14, 19, and 20 to give polycyclic compounds with 8-oxatricyclo[3.2.1.0(2,4)]octane and 9 oxatricyclo[3.3.1.0(2,4)]nonane frameworks. Generally, reactions proceed stereoselectively to give adducts of exo stereochemistry with the approach of the carbonyl ylide dipoles from the less-hindered face of cyclopropenes. The electronic properties of the substituent at the C3 position of cyclopropenes play an important role in governing the reactivity of cyclopropenes: when the C3 position is substituted by electron-acceptors such as the methoxycarbonyl or cyano groups, the yields of adducts are decreased significantly or no adducts can be detected at all. Relative reactivities of cyclopropenes were quantified by competition experiments to give the best correlation with sigmaF-Taft constants. Both measured photoelectron spectra and ground-state calculations of a series of 1,2-diphenylcyclopropenes indicate considerable lowering of cyclopropene pi-HOMO energies by substitution with an acceptor group. Such changes in electronic structures of cyclopropenes may cause the inversion of frontier molecular orbital (FMO) interactions from HOMO(cyclopropene)-LUMO(ylide) to LUMO(cyclopropene) HOMO(ylide) type. In terms of philicity, nucleophilic properties of acceptor substituted cyclopropenes are diminished to such an extent that these species are no longer good nucleophiles in the reaction with carbonyl ylides, and neither are they good electrophiles, being unreactive. This was shown by the B3LYP calculations of addends. PMID- 16709046 TI - Chemical reactivity as a tool to study carcinogenicity: reaction between chloroethylene oxide and guanine. AB - Chloroethylene oxide, an ultimate carcinogen of vinyl chloride, reacts with DNA giving rise to 7-(2-oxyethyl)guanine adduct in a nearly quantitative yield. This reaction represents an initial step of carcinogenesis associated with vinyl chloride. From experimental data for this reaction we calculated the second-order rate constant of 0.049 s(-1) M(-1), which corresponds to the activation free energy of 19.5 kcal/mol. We also performed a series of medium high ab initio and density functional theory simulations. Effects of hydration were considered in the framework of the Langevine dipoles solvation model and the solvent reaction field method of Tomasi and co-workers. In silico calculated activation free energies are in a good agreement with the experimental value. This fact presents strong evidence in favor of the validity of the proposed reaction mechanism and points to the applicability of quantum-chemical methods to studies of other reactions associated with carcinogenesis. Insignificant stereoselectivity of the studied reaction was also predicted. PMID- 16709047 TI - Synthesis and photophysical properties of dihydroheptacenes: new blue-emitting materials. AB - 7,16-Dihydroheptacenes (1-3) substituted at the 6, 8, 15, and 17 positions are synthesized as blue emitters potentially useful in organic light emitting diodes (OLEDs). The photophysical properties of 1-3 (lambda(max) = 424-428 nm, phi(F) = 0.15-0.21, tau(F) = 2.35-2.67 ns in CH2Cl2) are discussed. They are shown to be stable and efficient blue emitters in the solid state (phi(F) = 0.37-0.44). The X ray crystal structure of 1 is reported. PMID- 16709048 TI - Synthetic chlorins bearing auxochromes at the 3- and 13-positions. AB - Synthetic chlorins bearing diverse auxochromes at the 3- and 13-positions of the macrocycle are valuable targets given their resemblance to chlorophylls a and b, which bear 3-vinyl and 13-keto groups. A de novo route has been exploited to construct nine zinc chlorins bearing substituents at the 3- and 13-positions and two benchmark zinc chlorins lacking such substituents. The chlorins are sterically uncongested and bear (1) a geminal dimethyl group in the reduced pyrroline ring, (2) a H, an acetyl, a triisopropylsilylethynyl (TIPS-ethynyl), or a vinyl at the 3-position, (3) a H, an acetyl, or TIPS-ethynyl at the 13 position, and (4) a H or a mesityl at the 10-position. The synthesis of the 13 substituted chlorins relied on p-TsOH x H2O-catalyzed condensation of an 8,9 dibromo-1-formyldipyrromethane (eastern half) and 2,3,4,5-tetrahydro-1,3,3 trimethyldipyrrin (western half), followed by metal-mediated oxidative cyclization, affording the 13-bromochlorin. Similar use of a bromo- or TIPS ethynyl-substituted western half provided access to 3-substituted chlorins. A 3 bromo, 13-bromo, or 3,13-dibromochlorin was further transformed by Pd-coupling to introduce the vinyl group (via tributylvinyltin), TIPS-ethynyl group (via TIPS acetylene), or acetyl group (via tributyl(1-ethoxyvinyl)tin, followed by acidic hydrolysis). In the 10-mesityl-substituted zinc chlorins, the series of substituents, 3-vinyl, 13-TIPS-ethynyl, 3-TIPS-ethynyl, 13-acetyl, 3,13-bis(TIPS ethynyl), 3-TIPS-ethynyl-13-acetyl, or 3,13-diacetyl, progressively causes (1) a redshift in the absorption maximum of the B band (405-436 nm) and the Q(y) band (606-662 nm), (2) a relative increase in the intensity of the Q(y) band (I(B)/I(Q) = 4.2-1.5), and (3) an increase in the fluorescence quantum yield phi(f) (0.059-0.29). The zinc chlorins bearing a 3-TIPS-ethynyl-13-acetyl or a 3,13-diacetyl group exhibit a number of spectral properties resembling those of chlorophyll a or its zinc analogue. Taken together, this study provides access to finely tuned chlorins for spectroscopic studies and diverse applications. PMID- 16709049 TI - Optimization of diffusion-filtered NMR experiments for selective suppression of residual nondeuterated solvent and water signals from 1H NMR spectra of organic compounds. AB - Interpretation of 1H NMR spectra of organic compounds is sometimes hampered by the presence of strong peaks arising from residual nondeuterated solvent and water that obscure compound signals. Classical solvent suppression techniques such as presaturation or those based on pulsed field gradients are not effective in this regard because they also remove the compound resonances that overlap with the solvent signal being suppressed. Here, we propose an alternative scheme by using an optimized NMR diffusion filter that eliminates the nondesired peaks while retaining the signals of interest. This strategy has proved to be useful in three common deuterated solvents, namely, CDCl3, DMSO-d6, and CD3OD, resulting in clean spectra with no interference from solvent or water peaks. PMID- 16709050 TI - Water in oil microemulsions as reaction media for a Diels-Alder reaction between N-ethylmaleimide and cyclopentadiene. AB - The Diels-Alder reaction between N-ethylmaleimide and cyclopentadiene in water/AOT/isooctane microemulsions, where AOT denotes sodium bis(2 ethylhexyl)sulfosuccinate, was studied. The rate of the reaction was found to be higher than that obtained in pure isooctane, irrespective of the particular microemulsion composition used. The efficiency of this catalytic action ranged from a factor of 3 at low water contents (viz., W = [H2O]/[AOT] = 2) to 15 at W = 35. On the basis of these results, the reaction takes place simultaneously in the continuous medium and at the microemulsion interface. The favorable arrangement of the reactants at the interface results in more than 95% of the reaction occurring in this microenvironment. The kinetic analysis revealed the rate constant at the microemulsion interface to change with the water content. For small W values a bimolecular rate constant at the interface close to that observed in hexane was obtained. This value increases with W and for W > 20, a value close to that obtained in ethanol was found. This can be ascribed to the absence of hydrogen bonding at the microemulsion interface as well as the accelerating effects due to enforced hydrophobic interactions. PMID- 16709051 TI - Synthesis of (Z)-alkene and (E)-fluoroalkene-containing diketopiperazine mimetics utilizing organocopper-mediated reduction-alkylation and diastereoselectivity examination using DFT calculations. AB - We have carefully examined the organocopper-mediated reduction-alkylation of gamma-acetoxy or gamma,gamma-difluoro-alpha,beta-unsaturated-delta-lactams for the synthesis of (Z)-alkene- or (E)-fluoroalkene-containing diketopiperazine mimetics. Reduction of acetates 2, 12, 14, and difluorolactam 18 with higher order cuprate reagents (Me3CuLi2 x LiI x 3 LiBr), followed by trapping the resulting metal dienolate with an electrophile in a one-pot procedure gave alpha alkylated-beta,gamma-unsaturated-delta-lactams in good yields. Because of side chain steric repulsion, we found that alkylation using relatively large electrophiles such as BnBr gave mostly 3,6-trans isomers by kinetic trapping of metal enolates. On the other hand, MeI-mediated alkylations predominantly provided the unexpected 3,6-cis isomers despite the presence of a bulky benzyl side chain. Based on density functional theory calculations, we concluded that formation of the 3,6-cis isomers was due to the occurrence of oxa-pi-allyllithium complexes 29 and 31. PMID- 16709052 TI - Hydroxy-1-aminoindans and derivatives: preparation, stability, and reactivity. AB - The chemical stability and reactivity of hydroxy-1-aminoindans and their N propargyl derivatives are strongly affected by the position of the OH group and its orientation relative to that of the amino moiety. Thus, the 4- and 6-OH regioisomers were found to be stable, while the 5-OH analogues were found to be inherently unstable as the free bases. The latter, having a para orientation between the OH and the amino moieties, could be isolated only as their hydrochloride salts. 7-Hydroxy-1-aminoindans and 7-hydroxy-1-propargylaminoindans represent an intermediate case; while sufficiently stable even as free bases, they exhibit, under certain experimental conditions, unexpected reactivity. The instability of the 5- and 7-hydroxy-aminoindans is attributed to their facile conversion to the corresponding, reactive quinone methide (QM) intermediates. The o-QM obtained from 7-hydroxy-aminoindans was successfully trapped with ethyl vinyl ether via a Diels-Alder reaction to give tricyclic acetals 32a,b. PMID- 16709053 TI - Highly enantio- and diastereoselective Brassard type hetero-Diels-Alder approach to 5-methyl-containing alpha,beta-unsaturated delta-lactones. AB - Two efficient new chiral copper (II) Schiff base complexes were developed for the highly enantio- and diastereoselective HDA reaction of Brassard type diene 1b with aldehydes, to afford the corresponding 5-methyl-containing alpha,beta unsaturated delta-lactone derivatives in moderate yields, high enantioselectivities (up to 99% ee) and excellent diastereoselectivities (up to 99:1 anti/syn). On the basis of the absolute configuration of 4a-4j disclosed by X-ray diffraction and CD analysis, a possible transition-state model for the enantio- and diastereoselective catalytic reaction has been proposed. PMID- 16709054 TI - N,N'-carbonyldiimidazole-mediated cyclization of amino alcohols to substituted azetidines and other N-heterocycles. AB - Amino alcohols are important synthons for N-heterocycles. We have developed an efficient method to activate hydroxyl groups, which avoids the use of toxic reagents and tolerates a wide variety of functional groups. Our strategy has been applied to the synthesis of functionalized p-methoxyphenyl-protected azetidines, pyrrolidines, and piperidines. The required amino alcohols were synthesized according to an optimized proline-catalyzed Mannich protocol. An azetidine analogue of ezetimibe was synthesized to demonstrate the potential for the synthesis of drug-like molecules. PMID- 16709055 TI - Self-assembly of supramolecular platinum complexes with bis-4-pyridyl cavitands. AB - The design and self-assembly of six new supramolecular complexes (four triangles and two 2+2 assemblies) are described. These assemblies incorporate two new bispyridyl cavitand building blocks and were prepared in excellent yields (85 95%). The assemblies and building blocks were characterized with multinuclear NMR spectroscopy, electrospray ionization mass spectrometry, and elemental analysis. Isotopically resolved mass spectrometry along with NMR data confirms the existence of the six assemblies. PMID- 16709056 TI - Substrate-dependent dihydroxylation of substituted cyclopentenes: toward the syntheses of carbocyclic sinefungin and noraristeromycin. AB - Carbocyclic nucleosides are of considerable interest for the development of new therapeutic agents. A key reaction in the preparation of many such nucleoside analogues is dihydroxylation of appropriately substituted cyclopentenes. Although often considered a routine reaction, in this paper, we report the dramatic influence of substituents on the facial selectivity of dihydroxylations. The substituted cyclopentene substrates are derived from acylnitroso cycloaddition reactions of cyclopentadiene, followed by N-O reduction and efficient enzymatic resolution. The results are directly utilized in a very efficient asymmetric synthesis of an antiviral carbocyclic nucleoside, noraristeromycin 5. Extensions toward the synthesis of carbocyclic sinefungin 7 document the importance of realizing the substituent dependence of the dihydroxylation reaction. PMID- 16709058 TI - Distinguishing the early and late transition states and exploring the validity of sigma --> sigma*#, sigma# --> sigma*, and sigma --> pi*(C=O) concepts in diastereoselection from NBO analysis. AB - Natural bond orbital (NBO) analysis of several early TSs does not support the sigma --> sigma*# hypothesis. The sigma --> pi*(C=O) interaction controls the carbonyl pyramidalization that, in turn, controls the pi-selectivity of a nucleophilic addition. In contrast, late TSs are devoid of sigma --> pi*(C=O) interactions, and they benefit from sigma --> sigma*# interactions that control pi-selectivity. The evidence in favor of Anh-Felkin's sigma# --> sigma* hypothesis is weak. The electron-withdrawing sigma(C-F) in the 2-fluoropropanal LiCN TS did not align anti to the incipient bond even though there was complete conformational freedom. The initial guess for the TS in which sigma(C-F) was held anti to sigma# optimized to what had lost the said geometrical relationship. Furthermore, in the TS for axial addition of LiCN to 2-ax-F-cyclohexanone, the net sigma --> sigma*# interaction was considerably larger than the net sigma# --> sigma* interaction. The relative TS energies require that the equatorial addition of LiCN to 2-ax-F-cyclohexanone be favored over the axial addition in good compliance with the available experimental results. PMID- 16709057 TI - Copper(II)-catalyzed amidations of alkynyl bromides as a general synthesis of ynamides and Z-enamides. An intramolecular amidation for the synthesis of macrocyclic ynamides. AB - A general and efficient method for the coupling of a wide range of amides with alkynyl bromides is described here. This novel amidation reaction involves a catalytic protocol using copper(II) sulfate-pentahydrate and 1,10-phenanthroline to direct the sp-C-N bond formation, leading to a structurally diverse array of ynamides including macrocyclic ynamides via an intramolecular amidation. Given the surging interest in ynamide chemistry, this atom economical synthesis of ynamides should invoke further attention from the synthetic organic community. PMID- 16709059 TI - Improved synthesis of the A-G ring segment of brevetoxin B. AB - An efficient synthesis of the A-G ring segment 2, a key intermediate for the total synthesis of brevetoxin B (1), was achieved in 37 steps and 5.0% overall yield. The intramolecular allylation of the O,S-acetal 22, prepared from the ABC ring segment 15 and the FG ring segment 17, was carried out using AgOTf as a Lewis acid to give the desired compound 23, predominantly. Ring-closing metathesis of 23 with the Grubbs catalyst 12 afforded the heptacyclic ether 25. Selective hydrogenation of the E ring olefin of 25 was performed by diimide reduction to afford 2. PMID- 16709060 TI - Synthesis and hybridization studies of 2'-amino-alpha-L-LNA and tetracyclic "locked LNA". AB - A convergent route to a new class of locked nucleic acids, i.e., 2'-amino-alpha-L LNA, has been developed. The optimized synthetic route to the corresponding phosphoramidite building block of thymine proceeds in 4% overall yield over 15 steps from the starting diol. Crucial synthetic steps include (a) introduction of a C2-azido group prior to nucleobase coupling, (b) Vorbruggen glycosylation primarily affording the desired alpha-anomer, (c) separation of alpha-L-ribo- and beta-L-ribo-configured bicyclic nucleosides, and (d) selection of a suitable protecting group to avoid intramolecular Michael addition of the C2'-amino group onto the C6-position. Incorporation of a 2'-amino-alpha-L-LNA monomer into oligodeoxyribonucleotides results in modest changes in thermal stability with complementary DNA, whereas significant increases in thermal stability are observed with RNA complements along with excellent Watson-Crick discrimination. These results, along with the flexibility of the synthetic strategy allowing chemoselective N2'-functionalization at a late stage, render 2'-amino-alpha-L-LNA a promising building block for nucleic acid based nanobiotechnology and therapeutics. A slight modification in strategy facilitated the synthesis of the corresponding phosphoramidite building blocks of Michael adducts, which due to their tetracyclic skeletons exhibit a conformationally restricted furanose ring and glycosidic torsion angle (anti-range). Incorporation of such a "locked LNA" monomer into oligodeoxyribonucleotides results in large decreases in thermal affinity toward DNA/RNA complements. PMID- 16709061 TI - Stereoselective enzymatic synthesis of chiral alcohols with the use of a carbonyl reductase from Candida magnoliae with anti-Prelog enantioselectivity. AB - In our effort to search for carbonyl reductases with anti-Prelog enantioselectivity, the activity and enantioselectivity of a carbonyl reductase from Candida magnoliae have been examined with various ketones of diverse structures. This carbonyl reductase catalyzed the reduction of a series of ketones, alpha- and beta-ketoesters, to anti-Prelog configurated alcohols in excellent optical purity. The usefulness of this carbonyl reductase has been demonstrated by synthesis of several chiral alcohol intermediates of pharmaceutical importance. PMID- 16709062 TI - Photochemical cleavage and release of para-substituted phenols from alpha-keto amides. AB - In aqueous media alpha-keto amides 4-YC6H4OCH2COCON(R)CH(R')CH3 (5a, R = Et, R' = H; 5b, R = iPr, R' = Me) with para-substituted phenolic substituents (Y = CN, CF3, H) undergo photocleavage and release of 4-YC6H4OH with formation of 5 methyleneoxazolidin-4-ones 7a,b. For both 5a,b quantum yields range from 0.2 to 0.3. The proposed mechanism involves transfer of hydrogen from an N-alkyl group to the keto oxygen to produce zwitterionic intermediates 8a-c that eliminate the para-substituted phenolate leaving groups. The resultant imminium ions H2C=C(OH)CON+(R)=C(R')CH3 9a,b cyclize intramolecularly to give 7a,b. The quantum yields for photoelimination decrease in CH3CN, CH2Cl2, or C6H6 due to competing cyclization of 8a,b to give oxazolidin-4-one products which retain the leaving group 4-YC6H4O- (Y = H, CN). A greater tendency to undergo cyclization in nonaqueous media is observed for the N,N-diethyl amides 5a than the N,N diisopropyl amides 5b. With para electron releasing groups Y = CH3 and OCH3 quantum yields for photoelimination significantly decrease and 1,3 photorearrangment of the phenolic group is observed. The 1,3-rearrangement involves excited state ArO-C bond homolysis to give para-substituted phenoxyl radicals, which can be observed directly in laser flash photolysis experiments. PMID- 16709063 TI - Structure and synthesis of 6-(substituted-imidazol-1-yl)purines: versatile substrates for regiospecific alkylation and glycosylation at N9. AB - X-ray crystal structures of several 6-(azolyl)purine base and nucleoside derivatives show essentially coplanar conformations of the purine and appended 6 (azolyl) rings. However, the planes of the purine and imidazole rings are twisted approximately 57 degrees in a 2-chloro-6-(4,5-diphenylimidazol-1-yl)purine nucleoside, and a twist angle of approximately 61 degrees was measured between the planes of the purine and pyrrole rings in the structure of a 6-(2,5 dimethylpyrrol-1-yl)purine nucleoside derivative. Shielding "above" N7 of the purine ring by a proximal C-H on the 6-azolyl moiety is apparent with the coplanar compounds, but this effect is diminished in those without coplanarity. Syntheses of 6-(azolyl)purines from both base and nucleoside starting materials are described. Treatment of 2,6-dichloropurine with imidazole gave 2-chloro-6 (imidazol-1-yl)purine. Modified Appel reactions at C6 of trityl-protected hypoxanthine and guanine derivatives followed by detritylation gave 6-(imidazol-1 yl)- and 2-amino-6-(imidazol-1-yl)purines. Imidazole was introduced at C6 of 2',3',5'-tri-O-acetylinosine by a modified Appel reaction, and solvolysis of the glycosyl linkage gave 6-(imidazol-1-yl)purine. Guanosine triacetate was transformed into the protected 2,6-dichloropurine nucleoside, which was subjected to S(N)Ar displacement with imidazoles at C6 followed by glycosyl solvolysis to provide 2-chloro-6-(substituted-imidazol-1-yl)purines. Potential applications of these purine derivatives are outlined. PMID- 16709064 TI - Asymmetric synthesis of (-)-nupharamine and (-)-(5S,8R,9S)-5-(3-furyl)-8 methyloctahydroindolizidine from beta-amino ketones and the intramolecular Mannich reaction. AB - The asymmetric synthesis of the 2,3,6-trisubstituted piperidine core of the antitumor Nuphar alkaloids was readily achieved by using the intramolecular Mannich reaction and a sulfinimine-derived beta-amino ketone. PMID- 16709065 TI - A highly stereocontrolled total synthesis of (-)-neodysiherbaine A. AB - Neodysiherbaine A, a neuroexitotoxin occurring in a Micronesian marine sponge Dysidea herbacea, was synthesized from tri-O-acetyl-D-glucal with excellent stereocontrol. The method established enables us to obtain gram quantities of neodysiherbaine A and its related compounds. PMID- 16709066 TI - Synthesis of 1-arylmethyl-2-(cyanomethyl)aziridines and their ring transformation into methyl N-(2-cyanocyclopropyl)benzimidates. AB - 1-Arylmethyl-2-(cyanomethyl)aziridines were prepared in high yields from the corresponding 2-(bromomethyl)aziridines upon treatment with potassium cyanide in DMSO. Ring opening of the aziridine moiety with N-chlorosuccinimide in CCl4 and subsequent treatment of the thus formed 4-chloro-3-(N-chloro-N-(alpha,alpha dichlorobenzyl)amino)butanenitriles with sodium methoxide in methanol resulted in novel methyl N-(2-chloro-1-(cyanomethyl)ethyl)benzimidates, although in low yields. The latter gamma-chloro nitriles were smoothly converted into methyl N-(2 cyanocyclopropyl)benzimidates as precursors of biologically relevant beta-ACC derivatives through a 1,3-cyclization protocol by reaction with potassium tert butoxide in THF. PMID- 16709067 TI - A synthetic approach toward nitiol: construction of two 1,22-dihydroxynitianes. AB - Synthetic work toward the total synthesis of nitiol has culminated in the construction of two epimeric hydroxylated derivatives, the 1,22 dihydroxynitianes. Key stereodefining steps in the construction of the A-ring fragment (13) were the use of a siloxy-epoxide rearrangement reaction, a Pauson Khand reaction, a Norrish 1 photochemical cleavage reaction, and a highly regio- and stereoselective hydrostannylation reaction of an ynoate. The stereochemistry of the synthetically challenging C-ring fragment (20) was established using an Ireland-Claisen reaction and a Grubbs ring-closing metathesis process as key steps. The 12-membered B-ring of the nitiane skeleton was constructed using a copper-promoted Stille cross-coupling and a Kishi-Hiyama-Nozaki carbonyl addition reaction. Unfortunately, the carbonyl addition reaction produced hydroxyl functionality that could not be selectively removed. Consequently, a synthesis of epimeric 1,22-dihydroxynitianes, which are compounds that are structural hybrids of two natural products, nitiol and variculanol, was completed. PMID- 16709069 TI - Development of isomerization and cycloisomerization with use of a ruthenium hydride with N-heterocyclic carbene and its application to the synthesis of heterocycles. AB - A pure ruthenium hydride complex with N-heterocyclic carbene (NHC) ligand was efficiently generated from the reaction of a second-generation Grubbs ruthenium catalyst with vinyloxytrimethylsilane and unambiguously characterized. This ruthenium hydride complex showed high catalytic activity for the selective isomerization of terminal olefin and for the cycloisomerization of 1,6-dienes. These reactions of N-allyl-o-vinylaniline lead to novel synthetic methods for heterocycles such as indoles and 3-methylene-2,3-dihydroindoles, which are useful synthons for bioactive natural products. These procedures address an important issue in diversity-oriented synthesis. PMID- 16709068 TI - Acid-promoted reaction of the stilbene antioxidant resveratrol with nitrite ions: mild phenolic oxidation at the 4'-hydroxystiryl sector triggering nitration, dimerization, and aldehyde-forming routes. AB - In 0.1 M phosphate buffer, pH 3.0, and at 37 degrees C, resveratrol ((E)-3,4',5 trihydroxystilbene, 1a), an antioxidant and cancer chemopreventive phytoalexin, reacted smoothly at 25 microM or 1 mM concentration with excess nitrite ions (NO2(-)) to give a complex pattern of products, including two novel regioisomeric alpha-nitro (3a) and 3'-nitro (4) derivatives along with some (E)-3,4',5 trihydroxy-2,3'-dinitrostilbene (5), four oxidative breakdown products, 4 hydroxybenzaldehyde, 4-hydroxy-3-nitrobenzaldehyde, 3,5 dihydroxyphenylnitromethane, and 3,5-dihydroxybenzaldehyde, two dimers, the resveratrol (E)-dehydrodimer 6 and restrytisol B (7), and the partially cleaved dimer 2. The same products were formed in the absence of oxygen. 1H,15N HMBC and LC/MS analysis of the crude mixture obtained by reaction of 1a with Na (15)NO2 suggested the presence of 3,4',5,beta-tetrahydroxy-alpha-nitro-alpha,beta dihydrostilbene (8) as unstable intermediate which escaped isolation. Under similar conditions, the structurally related catecholic stilbene piceatannol ((E) 3,3',4,5'-tetrahydroxystilbene, 1b) gave, besides (E)-3,3',4,5'-tetrahydroxy-beta nitrostilbene (3b), 3,4-dihydroxybenzaldehyde and small amounts of 3,5 dihydroxybenzaldehyde. Mechanistic experiments were consistent with the initial generation of the phenoxyl radical of 1a at 4'-OH, which may undergo free radical coupling with NO2 at the alpha- or 3'-position, to give eventually nitrated derivatives and/or oxidative double bond fission products, or self-coupling, to give dimers. The oxygen-independent, NO2(-)-mediated oxidative fission of the double bond under mild, physiologically relevant conditions is unprecedented in stilbene chemistry and is proposed to involve breakdown of hydroxynitro(so) intermediates of the type 8. PMID- 16709070 TI - Solid-phase synthesis of oligodeoxyribonucleoside boranophosphates by the boranophosphotriester method. AB - Oligodeoxyribonucleoside boranophosphates (BH3-ODNs), containing four kinds of nucleobases, were synthesized by the solid-phase boranophosphotriester method. The 2'-deoxyribonucleoside 3'-boranophosphate monomers having 2-cyanoethyl (CE) groups as the phosphorus protecting groups were synthesized in good yields. A new condensing reagent, 1,3-dimethyl-2-(3-nitro-1,2,4-triazol-1-yl)-2-pyrrolidin-1-yl 1,3,2-diazaphospholidinium hexafluorophosphate, was found to be highly effective for the condensation reaction on the solid support. We also found that 1,8 bis(N,N-dimethylamino)naphthalene could accelerate the condensation reaction without causing beta-elimination of the CE groups from the boranophosphate triesters. The internucleotidic CE groups were selectively removed by treatment with 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) under anhydrous conditions. The acetylation of the terminal 5'-hydroxy group was found to be effective to suppress the decomposition of the BH3-ODNs during the DBU treatment on the solid support. Under optimized conditions for the solid-phase synthesis and the deprotection reactions, BH3-ODNs (4mers and 12mers) containing four kinds of nucleobases were synthesized in good yields. The hybridization properties of the BH3-ODN 12mers with the complementary native DNAs and RNAs were determined by the thermal denaturing studies. In contrast to the low thermal melting (Tm) value of the duplex composed of T((PB)T)11 and native dA12 (12.8 degrees C), the duplex consisting of d(C(PB)A(PB)G(PB)T)3 and d(ACTG)3 showed a higher Tm value (44.7 degrees C) under high-salt conditions. Furthermore, d(C(PB)A(PB)G(PB)T)3 formed a more stable duplex with the complementary RNA, r(ACUG)3 with a Tm value of 50.5 degrees C. Thus, we first demonstrated that the binding affinity of BH3-ODN to a complementary DNA or RNA is dramatically increased, owing to the inclusion of the four kinds of nucleobases. PMID- 16709071 TI - Palladium-catalyzed intramolecular asymmetric hydroamination, hydroalkoxylation, and hydrocarbonation of alkynes. AB - A conceptually novel approach for asymmetric intramolecular hydroamination, hydroalkoxylation and hydrocarbonation of alkynes using chiral palladium catalysts are described. The reactions of the aminoalkynes 5, alkynols 7, and alkynylmethines 9 in the presence of Pd2(dba)3 x CHCl3/PhCOOH/renorphos 4 in benzene (or benzene-hexane) at 100 degrees C gave the corresponding cyclization products (nitrogen heterocycles 6, oxygen heterocycles 8, and carbocycles 10) in good yields with good enantioselectivities. The origins of enantioselectivities in the hydroamination reaction are discussed based on DFT computations. PMID- 16709072 TI - A new reaction pathway in the ester aminolysis catalyzed by glymes and crown ethers. AB - Butylaminolysis of p-nitrophenyl acetate in chlorobenzene in the presence of different kinds of phase-transfer catalysts (crown ethers and glymes) supports the existence of a reaction pathway exhibiting a first-order dependence on the concentration of the phase transfer catalyst and a second-order dependence on the concentration of butylamine. This novel reaction pathway must be included in the mechanism traditionally accepted for the catalysis by phase-transfer agents of aminolysis reactions in aprotic solvents. PMID- 16709073 TI - Optical spectra of protected diamine 10-bond-bridged intervalence radical cations related to N,N,N'N'-tetraalkylbenzidine. AB - The optical absorption spectra of the delocalized intervalence radical cations of seven o,o'-linked benzidine derivatives that have the nitrogens protected as 9-(9 aza-bicyclo[3.3.1]nonan-3-one) derivatives are discussed and compared with that of the p-phenylene radical cation. The linking units are CH2, CH2CH2, NMe, S, SO2, and C=O, and we also studied H,H (the unlinked benzidine). The lowest-energy absorption band is assigned as the transition from the antibonding combination of symmetrical N and aromatic orbitals to the antibonding combination of the antisymmetric N and aromatic orbitals using TD-DFT calculations, and a good correlation between the observed transition energies and those calculated using the simple Koopmans theorem-based "neutral in-cation geometry" calculations on the UB3LYP/6-31G* structures is found. The use of the two-state model that equates the electronic interaction through the bridge between the amino groups with half of the lowest transition energy is seriously incorrect for these and other delocalized intervalence compounds. The problem of extracting the electronic interactions that actually are involved from calculated transition energies is discussed. PMID- 16709074 TI - An expeditious enantioselective synthesis of antimycin A3b. AB - A straightforward enantioselective route to (+)-antimycin A3b is presented, which used a TiCl4-mediated asymmetric aldolization to construct C-7/C-8 and BnOH/DMAP to remove the chiral auxiliary with concurrent protection of the carboxylic group, respectively. Closing the dilactone ring was achieved in 62% yield (previously 0.8%, 13.4%, or 20%) in the presence of the C-8 ester functionality. The overall yield (34.5%) was significantly higher than that (0.019-3.6%) of the earlier routes. PMID- 16709075 TI - Simple method for the preparation of esters from Grignard reagents and alkyl 1 imidazolecarboxylates. AB - The reaction of Grignard reagents with alkyl imidazolecarboxylates, which were prepared from alcohols with carbonyl diimidazole, gave the corresponding esters in good to excellent yields. This method conveniently provides esters from alkyl halides and alcohols by C1-carbon chain extension. PMID- 16709076 TI - Chiral seleno-amines from indium selenolates. A straightforward synthesis of selenocysteine derivatives. AB - A simple and efficient procedure for the synthesis of chiral beta-seleno-amines derivatives from a one-pot indium(I) iodide-mediated aziridine ring opening with diorganoyl diselenides has been developed. As an application, the synthesis of selenocysteine and selenotreonine derivatives has been accomplished. PMID- 16709077 TI - Detecting Hg2+ ions with an ICT fluorescent sensor molecule: remarkable emission spectra shift and unique selectivity. AB - A fluorescent ratiometric Hg2+ ion sensor RMS, based on a coumarin platform coupled with a tetraamide receptor, is presented. This sensor, employing the ICT mechanism, could be used to specifically detect Hg2+ ions in a neutral buffered water solution with an approximately 100-nm blue shift in emission spectra. PMID- 16709078 TI - Microwave-assisted one-pot synthesis of 1-indanones from arenes and alpha,beta unsaturated acyl chlorides. AB - A series of 1-indanones were synthesized in good yields via tandem Friedel-Crafts acylation and Nazarov cyclization of arenes and alpha,beta-unsaturated acyl chlorides in the presence of aluminum chloride under microwave irradiation. PMID- 16709079 TI - First stereospecific synthesis of (E)- or (Z)-alpha-fluoroenones via a kinetically controlled Negishi coupling reaction. AB - A highly stereospecific synthesis of (E)- or (Z)-alpha-fluoro-alpha,beta unsaturated ketones 4, via a kinetically controlled Negishi palladium-catalyzed coupling reaction, was developed, providing an easy and general access to valuable fluorinated intermediates (pharmaceutical, peptide mimic, and so on). The synthesis involved a reaction between E/Z gem-bromofluoroolefins 2 and alkoxyvinylzinc species 6 under controlled reaction temperature. At 10 degrees C, (Z)-4 (70 to 99% yields) was obtained along with unreacted (Z)-2 (66 to 99% yields). At THF reflux, the recovered olefin was transformed into (E)-4 (up to 98% yield). PMID- 16709080 TI - Diastereoselective arylithium addition to an alpha-trifluoromethyl imine. Practical synthesis of a potent cathepsin K inhibitor. AB - A practical, chromatography-free synthesis of potent cathepsin K inhibitor 1 is described. The addition of 4-bromophenyllithium to an alpha-trifluoromethylimine derived from commercially available (S)-leucinol was accomplished in a highly diastereoselective manner (97.6% de, 91% yield). Subsequent Suzuki cross-coupling afforded biaryl 7. Oxidation of the alcohol and sulfide functionalities led to the formation of carboxylic acid 8. Crystallization of 7 and acid 8 as its dicyclohexylamine salt gave excellent impurity rejection. The final amide coupling with commercially available aminoacetonitrile hydrochloride afforded 1 in excellent purity (99.6A% by HPLC, 100% de, <3 ppm Pd, W, Cr). PMID- 16709081 TI - Novel ambiphilic dichlorocarbenoid equivalent in alkene cyclopropanation and carbonyl olefination. AB - The Ti-Mg-dichloromethylene complexes derived from the oxidative addition of CCl4 to the Mg-TiCl4 bimetallic species serve as a novel class of ambiphilic dichlorocarbenoid equivalents. Not only is Ti-Mg-dichlorocarbenoid highly selective but also it seems highly reactive in both alkene cyclopropanations and carbonyl dichloromethylenations. PMID- 16709082 TI - Synthesis of 1-formyldipyrromethanes. AB - 1-Formyldipyrromethanes are versatile precursors to porphyrins and chlorins. Two methods of synthesis of 1-formyldipyrromethanes have been investigated: (1) Vilsmeier formylation followed by selective removal of the unwanted 1,9 diformyldipyrromethane by dialkyltin complexation and (2) reaction with mesitylmagnesium bromide (MesMgBr) followed by formylation with phenyl formate. The two approaches are complementary (acidic versus basic conditions; statistical versus selective formylation). The latter was found to be more efficient for the preparation of 1-formyldipyrromethanes. PMID- 16709083 TI - Synthesis of fluorene-based oligomeric organoboron reagents via Kumada, Heck, and Stille cross-coupling reactions. AB - Boronic pinacol ester group is not reactive in Kumada, Heck and Stille coupling reaction conditions. Fluorene-based sophisticated organoboron compounds were synthesized by means of Palladium catalyzed Kumada, Heck and Stille cross coupling reactions from halofluorenyl boronic esters. PMID- 16709084 TI - Efficient and practical synthesis of (R)-2-methylpyrrolidine. AB - An efficient, practical, and high yielding synthesis of (R)-2-methylpyrrolidine is described. The sequence allows for the scalable preparation of the target compound in just four synthetic steps and proceeds in 83% overall yield and >99% optical purity from readily available starting materials. PMID- 16709085 TI - Catalysis and regioselectivity of the aqueous Heck reaction by Pd(0) nanoparticles under ultrasonic irradiation. AB - An aqueous Heck reaction carried out under ultrasonic irradiation at the ambient temperature (25 degrees C) has been shown in this study to afford high yields of corresponding products. It was found that as a catalyst for the reaction palladium forms nanoparticles in-situ, characterized by transmission electron microscopy (TEM) and X-ray powder diffraction (XRD) analyses, and can be recycled. Furthermore, the Heck reaction under such mild and environmentally friendly conditions offers excellent regioselectivity of para- over ortho substitution in phenyl iodides especially with electron-donating groups. PMID- 16709086 TI - KO(t)Bu-promoted three-component coupling of aldehydes and alkynes: highly efficient synthesis of 1-en-4-yn-3-ols and 2-en-4-yn-1-ols. AB - A KO(t)Bu-promoted three-component coupling reaction of aldehydes and alkynes without a transition-metal catalyst was developed in which the sequential addition/isomerization/addition processes take place in one pot. This reaction could be developed into a straightforward and effective method for rapid access to stereodefined (E)-1-en-4-yn-3-ol and (Z)-2-en-4-yn-1-ol compounds. PMID- 16709087 TI - Novel cycloaddition reaction of [60]fullerene with carbonyl ylides generated from epoxides. AB - The thermal reaction of [60]fullerene (C60) with the carbonyl ylides generated in situ from trans-epoxides to give C60-fused tetrahydrofuran derivatives has been investigated. The reaction of C60 with trans-2-benzoyl-3-aryloxiranes afforded only cis-products, while the reaction of C60 with 2-cyano-2-ethoxycarbonyl-3 aryloxiranes gave exclusively or predominantly cis isomers. The isomeric distributions of the latter reactions were drastically affected by the substituent on the phenyl ring. PMID- 16709088 TI - One-pot preparation of arylalkynes by a tandem catalytic iodination of arenes and palladium-catalyzed coupling of iodoarenes with terminal alkynes. AB - Iodination of activated arenes by air-oxidation is carried out in the presence of catalytic bismuth salts at room temperature. Subsequently, the formed iodoarenes react with terminal alkynes to give arylalkynes under a selected palladium catalyzed coupling condition in the same pot. PMID- 16709090 TI - Nanotube-metal junctions: 2- and 3-terminal electrical transport. AB - We address the quality of electrical contact between carbon nanotubes and metallic electrodes by performing first-principles calculations for the electron transmission through ideal 2- and 3-terminal junctions, thus revealing the physical limit of tube-metal conduction. The structural model constructed involves surrounding the tube by the metal atoms of the electrode as in most experiments; we consider metallic (5,5) and n-doped semiconducting (10,0) tubes surrounded by Au or Pd. In the case of metallic tubes, the contact conductance is shown to approach the ideal 4e2/h in the limit of large contact area. For three terminals, the division of flux among the different transmission channels depends strongly on the metal material. A Pd electrode has nearly perfect tube-electrode transmission and therefore turns off the straight transport along the tube. Our results are in good agreement with some recent experimental reports and clarify a fundamental discrepancy between theory and experiment. PMID- 16709089 TI - Selective and reversible control of a chemical reaction with narrow-band infrared radiation: HXeCC radical in solid xenon. AB - The light-induced H + XeC2 <--> HXeCC reaction is studied in solid Xe, and the full optical control of this reaction is demonstrated. By narrow-band excitation in the IR spectral region, HXeCC radicals can be decomposed to a local metastable configuration and then selectively recovered by resonant excitation of the XeC2 vibrations. The novel recovery process is explained by short-range mobility of the reagents promoted by vibrational energy redistribution near the absorbing XeC2 molecule. This means that a chemical reaction can be selectively promoted in a desired place where the chosen absorber locates. The obtained results make a strong case of solid-state reactive vibrational excitation spectroscopy of weak radiationless transitions. PMID- 16709091 TI - Coarse-graining limits in open and wall-bounded dissipative particle dynamics systems. AB - Coarse graining of dense liquid-state systems can potentially lead to fast simulation times, thus providing an effective bridge between atomistic and continuum descriptions. Dissipative particle dynamics (DPD) is a stochastic Lagrangian method that provides a simple formal procedure for coarse graining. Here we analyze some of the fundamental modeling ideas of DPD and identify three factors that limit its application at high coarse-graining levels: interparticle force magnitude, compressibility, and geometric confinement. These artifacts lead to erroneous transport properties of highly coarse-grained DPD systems and thus incorrect dynamics in simulating complex fluids, e.g., colloids and polymers. PMID- 16709092 TI - Is the uniform electron gas limit important for small Ag clusters? Assessment of different density functionals for Ag(n) (n < or = 4). AB - Twenty-three density functional theory (DFT) methods, including the second- and the third-generation functionals, are tested in conjunction with two basis sets (LANL2DZ and SDD) for studying the properties of neutral and ionic silver clusters. We find that DFT methods incorporating the uniform electron gas limit in the correlation functional, namely, those with Perdew's correlation functionals (PW91, PBE, P86, and TPSS), Becke's B95, and the Van Voorhis-Scuseria functional VSXC, generally perform better than the other group of functionals, e.g., those incorporating the LYP correlation functional and variations of the B97 functional. Strikingly, these two groups of functionals can produce qualitatively different results for the Ag3 and Ag4 clusters. The energetic properties and vibrational frequencies of Ag(n) are also evaluated by the different functionals. The present study shows that the choice of DFT methods for heavy metals may be critical. It is found that the exact-exchange-incorporated PBE functional (PBE1PBE) is among the best for predicting the range of properties. PMID- 16709094 TI - Nose-Hoover dynamics in a shaker. AB - We introduce a new class of systems based on the Nose-Hoover equations. We show that we can add time-dependent terms without destroying the measure and energy conservation properties of the initial system. These "shakers" are typically pseudoperiodic in time, i.e., depend on a collection of harmonic oscillators. We show by numerical examples that it strengthens the sampling properties of the initial system with respect to the Gibbs measure and helps the computation of averages in the canonical ensemble. PMID- 16709093 TI - A polarizable continuum approach for the study of heterogeneous dielectric environments. AB - We present a computational method, exploiting some features of the polarizable continuum model (PCM) to describe heterogeneous media; it belongs to the family of electrostatic embedding mixed methods, such as the more common quantum mechanical (QM)/molecular mechanics approaches, with the electrostatic long range effects accounted for by a polarized continuum instead of atomic point charges. Provided effective dielectric constants are determined for the various parts of the system, the method is much faster than its atomistic counterpart, and allows for high-level QM calculations on the fragment of interest, using all the highly efficient computational tools developed for homogeneous PCM. Two case studies (the calculation of the pKa of solvent exposed acidic residues in a model protein, and the calculation of the electron spin resonance spectrum of a typical spin probe partially embedded in a membrane) are analyzed in some detail, to illustrate the application of the method to complex systems. PMID- 16709095 TI - Investigation of the dynamics of two coupled oscillators with mixed quantum classical methods. AB - The dynamics of two coupled oscillators can become quite complex if anharmonic potential energy functions are employed. This type of system therefore represents a good model for an investigation of the performance of mixed quantum-classical methods. In this work, the motion of two coupled particles with a mass ratio of one to ten is studied with three different mixed quantum-classical methods in the presence of anharmonic potential terms for a comparison with exact quantum mechanical calculations. The mixed quantum-classical approaches include the multitrajectory Ehrenfest, the mixed quantum-classical Bohmian (MQCB), and the so called coupled Schrodinger equations (CSE) formalisms. The analysis shows that while the description of a weakly anharmonic system by the Ehrenfest and MQCB schemes is accurate if proper sampling techniques are applied, both approximations break down rapidly if the anharmonic terms are increased. The performance of the simple CSE prescription, which corresponds to a reduction of the full two-dimensional wave function to two one-dimensional wave functions representing two quantum oscillators coupled via the potential energy in a classical fashion, decreases if the width of the initial wave packet is enlarged. The dependence of the CSE method on the diffuseness of the initial wave packet is therefore opposite to that of the MQCB method, which is more accurate for wide wave packets. Overall, the multitrajectory Ehrenfest ansatz is found to be most successful in reproducing the exact quantum results. PMID- 16709096 TI - Direct calculation of solid-vapor coexistence points by thermodynamic integration: application to single component and binary systems. AB - We present a new thermodynamic integration method that directly connects the vapor and solid phases by a reversible path. The thermodynamic integration in the isothermal-isobaric ensemble yields the Gibbs free energy difference between the two phases, from which the sublimation temperature can be easily calculated. The method extends to the binary mixture without any modification to the integration path simply by employing the isothermal-isobaric semigrand ensemble. The thermodynamic integration, in this case, yields the chemical potential difference between the solid and vapor phases for one of the components, from which the binary sublimation temperature can be calculated. The coexistence temperatures predicted by our method agree well with those in the literature for single component and binary Lennard-Jones systems. PMID- 16709097 TI - Efficient partitioning technique for computing the dynamics of intramolecular processes: radiationless transitions in pyrazine. AB - An efficient QP partitioning algorithm to compute the eigenvalues, eigenvectors, and the dynamics of large molecular systems of a particular type is presented. Compared to straightforward diagonalization, the algorithm displays favorable scaling (proportional to N(T)2) as a function of N(T), the size of the Hamiltonian matrix. In addition, the algorithm is trivially parallelizable, necessitating no "cross-talk" between nodes, thus enjoying the full linear speedup of parallelization. Moreover, the method requires very modest storage space, even for extremely large matrices. The method has also been enhanced through the development of a coarse-grained approximation, enabling an increase of the basis set size to unprecedented levels (10(8)-10(10) in the current application). The QP algorithm is applied to the dynamics of electronic internal conversion in a 24 vibrational-mode model of pyrazine. A performance comparison with other dynamical methods is presented, along with results for the decay dynamics of pyrazine and a discussion of resonance line shapes. PMID- 16709098 TI - A dual-level approach to density-functional theory. AB - An efficient approximate scheme for density-functional theory (DFT) calculations, which eliminates the time-consuming self-consistent-field (SCF) procedure, is proposed using a dual-level DFT approach. In this approach, dual levels of basis sets and exchange-correlation functionals are adopted. The dual-level DFT approach is based on the idea that the total electron density in the ground state can be represented in terms of the density evaluated using the low-quality basis set and the low-cost exchange-correlation functional. Since the SCF procedure is avoided in the total energy evaluation, the dual-level DFT approach drastically reduces the computational cost. The applications of several dual-level DFT calculations to molecular systems show that our approach is more efficient than the self-consistent DFT approach with a moderate accuracy. PMID- 16709099 TI - The midpoint method for parallelization of particle simulations. AB - The evaluation of interactions between nearby particles constitutes the majority of the computational workload involved in classical molecular dynamics (MD) simulations. In this paper, we introduce a new method for the parallelization of range-limited particle interactions that proves particularly suitable to MD applications. Because it applies not only to pairwise interactions but also to interactions involving three or more particles, the method can be used for evaluation of both nonbonded and bonded forces in a MD simulation. It requires less interprocessor data transfer than traditional spatial decomposition methods at all but the lowest levels of parallelism. It gains an additional practical advantage in certain commonly used interprocessor communication networks by distributing the communication burden more evenly across network links and by decreasing the associated latency. When used to parallelize MD, it further reduces communication requirements by allowing the computations associated with short-range nonbonded interactions, long-range electrostatics, bonded interactions, and particle migration to use much of the same communicated data. We also introduce certain variants of this method that can significantly improve the balance of computational load across processors. PMID- 16709100 TI - Hydration of ion-biomolecule complexes: ab initio calculations and gas-phase vibrational spectroscopy of K+ (indole)m(H2O)n. AB - In recent years neutral indole(H2O)n clusters have been used to model the hydration of biomolecules containing an indole moiety. Both experimental and theoretical studies of the binary indole...OH2 system show NH...OH sigma hydrogen bonding. By introducing a cation to the indole...OH2 model, cation...pi and ion...dipole electrostatic interactions are placed in direct competition with conventional indole...OH2 hydrogen-bonding. The effects, arising from a monovalent potassium cation on (indole)m(H2O)n clusters, were investigated using infrared photodissociation spectroscopy in the OH and NH stretching regions. In K+ (indole)1(H2O)(n < or = 4) and K+ (indole)2(H2O)(m < or = 3) clusters, the electrostatic ion...ligand interaction inhibits the formation of an indole NH...OH2 sigma hydrogen-bond. However, indole...H2O pi hydrogen-bonding via the five-membered indole ring is observed with three or more ligands around the ion. PMID- 16709101 TI - Excited states in the multireference state-specific coupled-cluster theory with the complete active space reference. AB - The recently proposed multireference state-specific coupled-cluster theory with the complete active space reference has been used to study electronically excited states with different spatial and spin symmetries. The algorithm for the method has been obtained using the computerized approach for automatic generation of coupled-cluster diagrams with an arbitrary level of the electronic excitation from a formal reference determinant. The formal reference is also used to generate the genuine reference state in the form of a linear combination of determinants contracted to a configuration with the spin and spatial symmetries of the target state. The natural-orbital expansions of the one-electron configuration inferaction density matrix allowed us to obtain the most compact orbital space for the expansion of the reference function. We applied our approach in the calculations of singlet and triplet states of different spatial symmetries of the water molecule. The comparisons of the results with values obtained using other many-particle methods and with the full configuration interaction results demonstrate good ability of the approach to deal with electronic excited states. PMID- 16709102 TI - A study of conical intersections for the H3(+) system. AB - A parametrization of the three asymptotic conical intersections between the energies of the H3(+) ground state and the first excited singlet state is presented. The influence of an additional, fourth conical intersection between the first and second excited states at the equilateral geometry on the connection between the three conical regions is studied, for both diatomics-in-molecules and ab initio molecular data. PMID- 16709103 TI - The Na2 2 3pi(g) state: new observations and hyperfine structure. AB - Many more Na2 2 3pi(g) v = 0-43, omega = 0, 1, 2 levels have been observed by sub Doppler continuous wave perturbation facilitated optical-optical double resonance fluorescence excitation spectroscopy and the hyperfine structure of the omega = 0 and 2 levels has been resolved. New molecular constants for the less perturbed v = 0-43 levels have been obtained with these new and the previously reported data. The hyperfine coupling scheme of the observed 2 3pi(g) levels is close to Hund's case a(beta) with a Fermi contact constant b(F) = 160+/-5 MHz, which is smaller than the Fermi contact constants of other Na2 triplet Rydberg states, b(F) = 200 245 MHz. PMID- 16709104 TI - Ab initio calculations of nonlinear optical rotation by several small chiral molecules and by uridine stereoisomers. AB - Expressions for nonlinear optical rotation are presented based on the quantum theory of optical birefringence of Atkins and Barron [Proc. R. Soc. London, Ser. A 304, 303 (1968); 306, 119 (1968)]. As concrete examples, the ordinary and nonlinear optical rotations are calculated with density functional theory (DFT) methodology for some simple single-ring molecules, namely, oxaziridine, diaziridine, and their derivatives, and for two, somewhat more complicated, conformations of uridine. For the single-ring molecules, (1) the angles of the ordinary optical rotation are mostly positive and (2) the contributions of the nonlinear effect to the total optical rotation depend both on the nature of the substituted species and of the host atom located on the ring. For the two conformations of uridine, (1) the signs of nonlinear optical rotation differ even though their ordinary optical rotations have the same sign and (2) whether the molecular structures are geometrically optimized with Hartree-Fock or DFT methodologies has no significant effect on the calculated nonlinear optical rotation when gauge-including atomic orbitals were used, even though the basis sets are small. These studies are expected to be helpful for interpretation of experimental results on nonlinear optical rotation by molecules underway in our research group. PMID- 16709105 TI - Electronic structures and chemical bonding in transition metal monosilicides MSi (M = 3d, 4d, 5d elements). AB - Bond distances, vibrational frequencies, electron affinities, ionization potentials, dissociation energies, and dipole moments of the title molecules in neutral, positively, and negatively charged ions were studied using the density functional method. Ground state was assigned for each species. The bonding patterns were analyzed and compared with both the available data and across the series. It was found that besides an ionic component, covalent bonds are formed between the metal s,d orbitals and the silicon 3p orbital. The covalent character increases from ScSi (YSi) to NiSi (PdSi) for 3d (4d) metal monosilicides, then decreases. For 5d metal monosilicides, the covalent character increases from LaSi to OsSi, then decreases. For the dissociation of cations, the dissociation channel depends on the magnitude of the ionization potential between metal and silicon. If the ionization potential of the metal is smaller than that of silicon, channel MSi+ --> M+ + Si is favored. Otherwise, MSi+ --> M + Si+ will be favored. A similar behavior was observed for anions, in which the dissociation channel depends on the magnitude of electron affinity. PMID- 16709106 TI - A molecular-beam optical Stark study of lines in the (1,0) band of the F4delta(7/2)-X4delta(7/2) transition of iron monohydride, FeH. AB - A supersonic molecular beam of iron monohydride, FeH, has been generated using a laser ablation/chemical reaction scheme and probed at near-natural linewidth resolution by optical Stark spectroscopy utilizing laser-induced fluorescence detection. The observed Stark splitting in Q(3.5) and R(3.5) lines of the F4delta(7/2) <-- X4delta(7/2) (1,0) transition were analyzed to determine values for the magnitudes of the permanent electric dipole moments, absolute value(mu), which were found to be 2.63(3) and 1.29(3) D for the X4delta (v = 0) and F4delta (v = 1) states, respectively. A comparison with ab initio theoretical predictions is made. The lambda doubling in the low-J levels of the F4delta(7/2) (v = 1) state is also modeled. PMID- 16709107 TI - The quadrupole moment of the Sb nucleus from molecular microwave data and calculated relativistic electric-field gradients. AB - The recently determined accurate values of the nuclear quadrupole coupling constant of the Sb nucleus in SbN, SbP, SbF, and SbCl and the calculated electric field gradients at Sb in these molecules are used to obtain the nuclear quadrupole moment of 121Sb and 123Sb. The calculation of the electric-field gradient has been carried out by using the infinite-order two-component relativistic method in the scalar approximation. The accompanying change of picture of the electric-field gradient operator has been accounted for by employing the shifted nucleus model of nuclear quadrupoles. The electron correlation effects are calculated at the level of the coupled cluster approximation. The present calculations give the "molecular" value of the nuclear quadrupole moment of 121Sb equal to -556+/-24 mb which is considerably different from the old "recommended" value of -360+/-40 mb and also differs from the recent "solid-state" result (-669+/-15 mb). The validation of the present data is comprehensively discussed. PMID- 16709108 TI - Search for global-minimum geometries of medium-sized germanium clusters. II. Motif-based low-lying clusters Ge21-Ge29. AB - We performed a constrained search for the geometries of low-lying neutral germanium clusters Ge(N) in the size range of 21 < or = N < or = 29. The basin hopping global optimization method is employed for the search. The potential energy surface is computed based on the plane-wave pseudopotential density functional theory. A new series of low-lying clusters is found on the basis of several generic structural motifs identified previously for silicon clusters [S. Yoo and X. C. Zeng, J. Chem. Phys. 124, 054304 (2006)] as well as for smaller sized germanium clusters [S. Bulusu et al., J. Chem. Phys. 122, 164305 (2005)]. Among the generic motifs examined, we found that two motifs stand out in producing most low-lying clusters, namely, the six/nine motif, a puckered hexagonal-ring Ge6 unit attached to a tricapped trigonal prism Ge9, and the six/ten motif, a puckered-hexagonal-ring Ge6 unit attached to a bicapped antiprism Ge10. The low-lying clusters obtained are all prolate in shape and their energies are appreciably lower than the near-spherical low-energy clusters. This result is consistent with the ion-mobility measurement in that medium-sized germanium clusters detected are all prolate in shape until the size N approximately 65. PMID- 16709109 TI - Converged vibrational energy levels and quantum mechanical vibrational partition function of ethane. AB - The vibrational partition function of ethane is calculated in the temperature range of 200-600 K using well-converged energy levels that were calculated by vibrational configuration interaction, and the results are compared to the harmonic oscillator partition function. This provides the first test of the harmonic oscillator approximation for a molecule with more than five atoms. The absolute free energies computed by the harmonic oscillator approximation are in error by 0.59-0.62 kcal/mol over the 200-600 K temperature range. PMID- 16709110 TI - Energetics of the manganese oxide cluster cations Mn(N)O+ (N = 2-5): role of oxygen in the binding of manganese atoms. AB - The photodissociation of manganese oxide cluster cations Mn(N)O+ (N = 2-5), into Mn(N-1)O+ (one-atom loss) and Mn(N-2)O+ (two-atom), was investigated in the photon-energy range of 1.08-2.76 eV. The bond-dissociation energies D0(Mn(N 1)O+...Mn) for N = 3, 4, and 5 were determined to be 1.84+/-0.03, 0.99+/-0.05, and 1.25+/-0.14 eV, respectively, from the threshold energies for the one- and two-atom losses. As Mn2O+ did not dissociate even at the highest photon energy used, the bond dissociation energy of Mn2O+, D0(Mn+...MnO), was obtained from a density-functional-theory calculation to be 3.04 eV. The present findings imply that the core ion Mn2O+ is bound weakly with the rest of the manganese atoms in Mn(N)O+. PMID- 16709111 TI - Infrared multiphoton ionization of superhot C60: experiment and model calculations. AB - We address, both experimentally and theoretically, the issue of infrared (IR) resonance enhanced multiphoton ionization (IR-REMPI) and thermally induced redshifts of IR absorption lines in a very large and highly vibrationally excited molecular system. Isolated superhot C60 molecules with well defined and variable average vibrational energy in the range of 9-19 eV, effusing out of a constant flux thermal source, are excited and ionized after the absorption of multiple (500-800) infrared photons in the 450-1800 cm(-1) spectral energy range. Recording the mass-selected ion signal as a function of IR wavelength gives well resolved IR-REMPI spectra, with zero off-resonance background signal. An enhancement of the ion signal of about a factor of 10 is observed when the temperature is increased from 1200 to 1800 K under otherwise identical conditions. A pronounced temperature dependent redshift of some of the IR absorption lines is observed. The observations are found to be in good agreement with a model which is based on the sequential absorption of single photons, always followed by instantaneous vibrational energy redistribution. The mass spectra (C60(+) fragmentation pattern) are found to be strongly excitation wavelength dependent. Extensive fragmentation down to C32(+) is observed following the absorption of 1350-1400 cm(-1) as well as 1500-1530 cm(-1) photons while negligible fragmentation is observed when exciting around 520 cm(-1). PMID- 16709112 TI - Observation of dihalide elimination upon electron attachment to oxalyl chloride and oxalyl bromide, 300-550 K. AB - Rate coefficients have been measured for electron attachment to oxalyl chloride [ClC(O)C(O)Cl] and oxalyl bromide [BrC(O)C(O)Br] in He gas at 133 Pa pressure over the temperature range of 300-550 K. With oxalyl chloride, the major ion product of attachment is Cl2(-) at all temperatures (66% at 300 K); its importance increases slightly as temperature increases. Two other product ions formed are Cl- (18% at 300 K) and the phosgene anion CCl2O- (16% at 300 K) and appear to arise from a common mechanism. With oxalyl bromide, the Br2(-) channel represents almost half of the ion product of attachment, independent of temperature. Br- accounts for the remainder. For oxalyl chloride, the attachment rate coefficient is small [(1.8 +/- 0.5) x 10(-8) cm3 s(-1) at 300 K], and increases with temperature. The attachment rate coefficient for oxalyl bromide [(1.3 +/- 0.4) x 10(-7) cm3 s(-1) at 300 K] is nearly collisional and increases only slightly with temperature. Stable parent anions C2Cl2O2(-) and C2Br2O2(-) and adduct anions Cl- (C2Cl2O2) and Br- (C2Br3O2) were observed but are not primary attachment products. G2 and G3 theories were applied to determine geometries of products and energetics of the electron attachment and ion-molecule reactions studied. Electron attachment to both oxalyl halide molecules leads to a shorter C-C bond and longer C-Cl bond in the anions formed. Trans and gauche conformers of the neutral and anionic oxalyl halide species have similar energies and are more stable than the cis conformer, which lies 100-200 meV higher in energy. For C2Cl2O2, C2Cl2O2(-), and C2Br2O2(-), the trans conformer is the most stable conformation. The calculations are ambiguous as to the oxalyl bromide geometry (trans or gauche), the result depending on the theoretical method and basis set. The cis conformers for C2Cl2O2 and C2Br2O2 are transition states. In contrast, the cis conformers of the anionic oxalyl halide molecules are stable, lying 131 meV above trans-C2Cl2O2(-) and 179 meV above trans-C2Br2O2(-). Chien et al. [J. Phys. Chem. A 103, 7918 (1999)] and Kim et al. [J. Chem. Phys. 122, 234313 (2005)] found that the potential energy surface for rotation about the C-C bond in C2Cl2O2 is "extremely flat." Our computational data indicate that the analogous torsional surfaces for C2Br2O2, C2Cl2O2(-), and C2Br2O2(-) are similarly flat. The electron affinity of oxalyl chloride, oxalyl bromide, and phosgene were calculated to be 1.91 eV (G3), and 2.00 eV (G2), and 1.17 eV (G3), respectively. PMID- 16709113 TI - Fragmentation dynamics of argon clusters (Ar(n), n = 2 to 11) following electron impact ionization: modeling and comparison with experiment. AB - The fragmentation dynamics of argon clusters ionized by electron impact is investigated for initial cluster sizes up to n = 11 atoms. The dynamics of the argon atoms is modeled using a mixed quantum-classical method in which the nuclei are treated classically and the transitions between electronic states quantum mechanically. The potential-energy surfaces are derived from a diatomics-in molecules model with the addition of the induced dipole-induced dipole and spin orbit interactions. The results show extensive and fast fragmentation. The dimer is the most abundant ionic fragment, with a proportion increasing from 66% for n = 2 to a maximum of 95% for n = 6 and then decreasing down to 67% for n = 11. The next abundant fragment is the monomer for n < 7 and the trimer otherwise. The parent ion dissociation lifetimes are all in the range of 1 ps. Long-lived trajectories appear for initial cluster sizes of seven and higher, and favor the formation of the larger fragments (trimers and tetramers). Our results show quantitative agreement with available experimental results concerning the extensive character of the fragmentation: Ar+ and Ar2(+) are the only ionic fragments for sizes up to five atoms; their overall proportion is in quantitative agreement for all the studied sizes; Ar2(+) is the main fragment for all sizes; stable Ar3(+) fragments only appear for n > or = 5, and their proportion increases smoothly with cluster size from there. However, the individual ionic monomer and dimer fragment proportions differ. The experimental ones exhibit oscillations with initial cluster size, with a slight tendency to decrease on average for the monomer. In contrast our results show a monotonic, systematic evolution, similar to what was found in our earlier studies on neon and krypton clusters. Several hypotheses are discussed in order to find the origin of this discrepancy. Finally, the metastable II(1/2)u and II(1/2)g states of Ar2(+) are found to decay with a lifetime of 3.5 and 0.1 ps, respectively, due to spin-orbit coupling. The difference with the commonly accepted microsecond range value for rare-gas dimer ions could originate from the role of autoionizing states in the formation of the parent ions. PMID- 16709114 TI - Quasiclassical trajectory study of O(1D) + N2O --> NO + NO: classification of reaction paths and vibrational distribution. AB - Quasiclassical trajectory calculations for the planar reaction of O(1D) + N2O --> NO + NO are performed on a newly constructed ab initio potential energy surface. In spite of the reduced dimension approximation, the agreement between the computational and experimental results is largely satisfactory, especially on the similar amount of excitation of the two kinds of NO products found by Akagi et al. [J. Chem. Phys. 111, 115 (1999)]. Analyzing the initial condition dependence of the trajectories, we find that the trajectories of this reaction can be classified into four reaction paths, which correspond to respective areas in the space of initial condition. In one of the four paths, a long-lived stable complex is formed in the course of reaction, whereas the other three paths have direct mechanism. Contradictory to conventional understanding of the chemical reaction dynamics, the direct paths show more efficient energy exchange between the NO stretching modes than that with a long-lived intermediate. This indicates that the vibrational mode coupling along the short-lived paths is considerably stronger than expected. PMID- 16709115 TI - Theoretical study of structure and photoelectron spectroscopy of In(x)P(y)- and In(x)P(y) (x + y < or = 6) clusters. AB - The geometries and vibrational frequencies of In(x)P(y)- and In(x)P(y) are investigated by hybrid B3LYP functional for x + y < or = 6 and CCSD(T) method for x + y < or = 3. As for the small clusters having two to three atoms, the geometrical and electronic structures and vibrational frequencies at the B3LYP level are in good agreement with those at the CCSD(T) level. Among the most stable structures of In(x)P(y)- and In(x)P(y) (x + y < or = 6) clusters the P rich clusters are more stable than In-rich clusters. Moreover, we found that those P atoms in In(x)P(y)- and In(x)P(y) (x + y < or = 6) clusters prefer to form a P-P bond, triangle, quadrangle, and pentagon for y = 2, 3, 4, and 5, respectively. Also, the vertical detachment energies of In(x)P(y)- (x + y < or = 6) and electron affinities of In(x)P(y) (x + y < or = 6) clusters obtained by B3LYP are in good agreement with the experimental values available. Theoretically, we show that the electron affinity of In3P3 is very low because, as observed in the experiment, there is a formation of a new P-P bond after an electron is lost from In(3)P(3)-, and we find that the similar phenomena exhibit in In2P4(-) cluster as well. PMID- 16709116 TI - Structure of low-lying electronic states of NdO: quantum chemical calculations. AB - Low-lying states of the NdO molecule have been predicted from quantum mechanical complete active-space self-consistent field/multireference configuration interaction/spin-orbit calculations. 54 states labeled through the quantum number Omega(+/-) have been determined in the excitation energy range of approximately 1 eV. For each state molecular constants T(e), T(v), omega(e), deltaG(v), R(e), B(e), and B(v) have been calculated. All these states display nearly identical principal structural characteristics: equilibrium internuclear distance and vibrational frequency. Calculated values of T(v), deltaG(v), and B(v) agree satisfactorily with experimental values available for nine electronic states among the 54 considered. The feasibility of a statistical representation of the low-lying states of NdO is considered. PMID- 16709117 TI - Experimental and theoretical studies of lambda doublings and permanent electric dipoles in the low-lying 1pi states of NaCs. AB - We present experimental data on the electric permanent dipole moments d(v',J') and lambda splittings (q factors) in the quasidegenerate (3) 1pi(e/f) state of the NaCs molecule over a wide range of the vibrational (v') and rotational (J') quantum numbers by using the combination of dc Stark mixing and electric radio frequency-optical double resonance methods. Within the experimental (3) 1pi state v' ranged from v' = 0 to 34, q values exhibited a pronounced decrease from 7.91x10(-6) to 0.47x10(-6) cm(-1), while absolute value(d) values varied between 8.0 and 5.0 D. Experimental evaluation yielded small d values about 1 D for D2 1pi state v' < 3 levels. The experiment is supported by ab initio electronic structure calculations performed for the (1-3) 1pi states of NaCs by means of the many-body multipartitioning perturbation theory of potential energy curves, permanent dipole, and angular coupling matrix elements for the lowest singlet states. The predicted d values reproduce their experimental counterparts within the measurement errors while theoretical q factors reproduce the measured v' dependence being, however, systematically overestimated by ca. 1x10(-6) cm(-1). The present NaCs data are compared with those of the NaK and NaRb molecules. PMID- 16709118 TI - The geometric, electronic, and magnetic properties of Ag5X+ (X = Sc, Ti, V, Cr, Mn, Fe, Co, and Ni) clusters. AB - Density functional theory calculations are performed on small cationic transition metal doped silver clusters, Ag5X+ (X = Sc, Ti, V, Cr, Mn, Fe, Co, and Ni) using the B3LYP and BP86 functionals. Several two-dimensional and three-dimensional isomers with the dopant at a high coordinated site are found to be close in energy. The relative energy of the isomers is checked with CCSD(T). The interaction between the dopant 3d electrons and the host is discussed by considering the density of states and the shape of the molecular orbitals. A large local spin magnetic moment on the dopant atom is predicted. PMID- 16709119 TI - The spectrum of CH2 near 1.36 and 0.92 microm: reevaluation of rotational level structure and perturbations in a(010). AB - The spectrum of methylene in the 1.3-1.4 and 0.89-0.94 microm wavelength regions has been recorded in absorption using frequency-modulated cw diode and Ti:sapphire laser sources. The spectral lines have Doppler-limited resolution and have been assigned to bands in the b(1)B1 <-- a(1)A1 electronic spectrum of the radical. In three of the four bands studied, the lower state is the bend excited, v2" = 1, level of the a state and two of the upper levels lie below the energy of the degenerate linear configuration of the b/a pair. Together with previously measured data pertaining to v2" = 1, the data have been used to refine the precision of the experimentally determined rotational structure in this level. Although several K" = 1 levels do show shifts of more than 0.1-0.2 cm(-1), multiple strong perturbations due to near-resonant background X(3)B1 rovibrational levels, such as are known to occur in the a(1)A1, v2" = 0 level have not been found in v2" = 1. Absorption lines due to the predominantly triplet X(040) 4(14) level, responsible for most of the perturbation of a(010) 5(15), have been identified in the spectra. The data also fix the energies of the b(0,0,0)2, a(0,7,0)1, b(0,2,0)3, and a(0,10,0)2 upper vibronic levels, where the numbers in parentheses are the vibrational quantum numbers with superscript K, the projection of the total angular momentum on the a-inertial axis. PMID- 16709120 TI - Neutron reflectivity measurements of the translational motion of tris(naphthylbenzene) at the glass transition temperature. AB - The translational dynamics of the low molecular weight glass-former tris(naphthylbenzene) have been studied on the length scale of a few nanometers at the glass transition temperature Tg. Neutron reflectivity was used to measure isotopic interdiffusion of multilayer samples created by physical vapor deposition. Deposition with the substrate held at Tg-6 K allows observation of dynamics characterizing the equilibrium supercooled liquid. The diffusion coefficient measured at q = 0.03 A(-1) was determined to be 1x10(-17) cm2/s at 342 K (Tg). The self-part of the intermediate scattering function I(s)(q,t) decays exponentially. Samples deposited well below Tg show a substantial thermal history effect during subsequent translational motion at Tg. PMID- 16709121 TI - An ab initio study of the fcc and hcp structures of helium. AB - The hexagonal close packed (hcp) and face centered cubic (fcc) structures of helium are studied by using a new ab initio computational model for large complexes comprising small subsystems. The new model is formulated within the framework of the energy incremental scheme. In the calculation of intra- and intersystem energies, model systems are introduced. To each subsystem associated is a set of partner subsystems defined by a vicinity criterion. In the independent calculations of intra- and intersystem energies, the calculations are performed on model subsystems defined by the subsystems considered and their partner subsystems. A small and a large basis set are associated with each subsystem. For partner subsystems in a model system, the small basis set is adopted. By introducing a particular decomposition scheme, the intermolecular potential is written as a sum of effective one-body potentials. The binding energy per atom in an infinite crystal of atoms is the negative value of this one body potential. The one-body potentials for hcp and fcc structures are calculated for the following nearest neighbor distances (d0): 4.6, 5.1, 5.4, 5.435, 5.5, 5.61, and 6.1 a.u. The equilibrium distance is 5.44 a.u. for both structures. The equilibrium dimer distance is 5.61 a.u. For the larger distances, i.e., d0 > 5.4 a.u., the difference of the effective one-body potentials for the two structures is less than 0.2 microE(h). However, the hcp structure has the lowest effective one-body potential for all the distances considered. For the smallest distance the difference in the effective one-body potential is 3.9 microE(h). Hence, for solid helium, i.e., helium under high pressure, the hcp structure is the preferred one. The error in the calculated effective one-body potential for the distance d0 = 5.61 a.u. is of the order of 1 microE(h) (approximately 0.5%). PMID- 16709122 TI - Excitation wavelength dependence of the Raman-Stokes shift of N,N-dimethyl-p nitroaniline. AB - Raman spectra of N,N-dimethly-p-nitroaniline have been measured in various solvents. The Raman-Stokes shift of the band assigned to the NO2 stretching mode excited at 488 nm was found to be linearly dependent on the pi-pi* absorption band center. Furthermore, it is found that the Raman-Stokes shift of the NO2 stretching mode is dependent upon the excitation wavelength. The extent of the shift when excited at 355 versus 488 nm is almost linearly dependent on the vibrational bandwidth of the NO2 mode. The phenomenon is interpreted as the result of the solvation state selective excitation of the vibrational mode as in the case of phenol blue [Yamaguchi et al., J. Chem. Phys. 109, 9075 (1998); 109, 9084 (1998)]. PMID- 16709123 TI - Development of a new polarizable potential model of hydrogen fluoride and comparison with other effective models in liquid and supercritical states. AB - Development of a new polarizable potential of hydrogen fluoride through the reparametrization of the JV-P model is presented: The length of the H-F bond has been shortened and the other parameters of the model have been readjusted accordingly. The structural, thermodynamic, and liquid-vapor equilibrium properties of the new model are compared with those of other effective potential models of HF as well as with experimental data in a broad range of thermodynamic states, from near-freezing to supercritical conditions. It is found that although the reparametrization does not change the structural properties of the HF model noticeably at the level of the pair correlations, it improves the reproduction of the thermodynamic properties of hydrogen fluoride over the entire range of existence of a thermodynamically stable liquid phase and also that of the vapor liquid coexistence curve. However, the new model, which still overestimates the close-contact separation of the HF molecules, underestimates the density of the coexisting liquid phase and overestimates the saturation pressure, probably due to the too steep repulsion of the potential function. PMID- 16709124 TI - Contribution of the asymmetric stretch, nu3B1, to the fundamental Raman spectrum of water. AB - The OD-stretching overtone from liquid D2O, 2nu, and the fundamental OD stretch from dilute HDO, both display high-frequency depolarization ratio minima, but the fundamental OD stretch from neat D2O displays a maximum, at the equivalent position. The rhoL minima arises from the decreased depolarization ratio produced by the absence of B1 modes. The fundamentals of HDO are of A species, and the 2nu overtone of D2O only involves A1 species, e.g., 2nu3B1 has A1 species via B1 x B1 = A1. A and A1 modes display small rhoL values which produce minima in rhoL near 2665 cm(-1) for HDO, and near 5250 cm(-1) for the D2O overtone. These minima give way to a depolarization ratio maximum when the depolarized, rhoL = 34, nu3B1 fundamental, makes its appearance in D2O at 2650 cm(-1). Fundamental and overtone depolarization ratios were used to determine the nu3B1 contribution to the depolarization ratio of the fundamental OD stretch; a value of approximately 28% resulted at 2655 cm(-1). Liquid H2O displays completely analogous features; a value of approximately 20% resulted for it at 3660 cm(-1). Nonhydrogen-bonded nu3B1, and more strongly hydrogen-bonded nu3B1, modes are also indicated for D2O and H2O. A rigorous test of the current results can be accomplished by measuring the depolarization ratio of the extraordinarily weak second Raman overtone, 3nu, recently detected for D2O. PMID- 16709125 TI - Molecular dynamics simulations of nanoparticles in dense isotropic nematogens: the role of matrix-induced long-range repulsive interactions. AB - We have carried out molecular dynamics simulation studies of binary mixtures of spherical nanoparticles (NPs) in a matrix of dense isotropic rod-shaped nematogens, with the size of the nematogen length being similar to that of the NP diameter. NPs at even low concentrations were found to shift the isotropic nematic (I-N) transition significantly to higher pressure at a given temperature, indicative of long-range perturbation of the nematogenic matrix by the NPs. The NPs were found to be dispersed in the dense isotropic nematogenic matrix over a wide range of NP concentrations due to long-range (compared with the molecular size of the nematogens) repulsion caused by NP-induced local order fluctuations and reduced local orientational correlation in the isotropic nematogenic matrix, in contrast to the phase separation predicted and observed in other studies where the particles were much larger or smaller than the nematogens. Furthermore, since the repulsion observed in the NP-nematogen mixtures is only microscopically long range (on the order of about ten molecular lengths of the nematogens), globally ordered clustering observed in mixtures of colloidal particles in nematic matrices resulting from macroscopically long-range interaction is not observed in our simulations. PMID- 16709126 TI - Excitation of nanoscale vapor bubbles at the surface of gold nanoparticles in water. AB - Intense nonequilibrium femtosecond laser excitation of gold nanoparticles in water leads to a transient heating of the nanoparticles, which decays via heat transfer to the water phase. It is shown that the water temperature rises to near the critical temperature and the water undergoes an explosive evaporation in the subnanosecond range. The formation of vapor bubbles shows a threshold dependence on laser fluence. The nascent nanoscale vapor bubbles change the heat dissipation drastically. The nanoscale structure is resolved directly with a combination of x ray scattering methods sensitive to the particle lattice expansion and the change in the water structure factor. PMID- 16709127 TI - The generalized molecular fractionation with conjugate caps/molecular mechanics method for direct calculation of protein energy. AB - A generalized molecular fractionation with conjugate caps/molecular mechanics (GMFCC/MM) scheme is developed for efficient linear-scaling quantum mechanical calculation of protein energy. In this GMFCC/MM scheme, the interaction energy between neighboring residues as well as between non-neighboring residues that are spatially in close contact are computed by quantum mechanics while the rest of the interaction energy is computed by molecular mechanics. Numerical studies are carried out to calculate torsional energies of six polypeptides using the GMFCC/MM approach and the energies are shown to be in general good agreement with the full system quantum calculation. Among those we tested is a polypeptide containing 396 atoms whose energies are computed at the MP26-31G* level. Our study shows that using GMFCC/MM, it is possible to perform high level ab initio calculation such as MP2 for applications such as structural optimization of protein complex and molecular dynamics simulation. PMID- 16709128 TI - Density functional theory study of water dissociation in a double water bilayer with or without coadsorption of CO on Pt(111). AB - Using density functional theory, we investigate the structure of the double water bilayer with or without coadsorption of CO on Pt(111). The double water bilayer consists of two bilayers. Each bilayer is buckled with every second water molecule being closer to the surface than every other water molecule. CO is found to adsorb most strongly when substituting in the first bilayer, the water molecule closest to the surface. Dissociation of H2O in the water bilayer (with or without CO) is further considered. A great number of pathways for the dissociation are studied. These include homolytic pathways where both dissociation products end up adsorbed on the Pt surface and heterolytic pathways where only the OH is adsorbed, while a proton is transferred to the water adlayers. We find that the heterolytic dissociation pathways are energetically more favorable than the homolytic ones, yet they are all rather endothermic. The most favorable pathways found have reaction energies of 0.60 and 0.52 eV without and with CO present. The corresponding activation energies are 0.99 and 0.53 eV, respectively. PMID- 16709129 TI - Monitoring the chemical vapor deposition growth of multiwalled carbon nanotubes by tapered element oscillating microbalance. AB - The growth of multiwalled carbon nanotubes (MWCNTs) produced by a catalytic chemical vapor deposition (CCVD) process has been monitored using a tapered element oscillating microbalance (TEOM) probe. This technique displays a high sensitivity (<1 microg). Growths in the TEOM microreactor are investigated with catalytic particles (Fe, Ni) dispersed on different supports. First, high surface area FeAl2O3 or Fe (Ni) exchanged on zeolite powders is used. Second, growths are performed on array of nickel dots or FeSi-nc particles dispersed on large holes patterned on Si(100) substrates. An accurate monitoring of the early stages of growth permits a precise evaluation of the growth rates and shows substantial differences between these samples which greatly differ by the surface area. On catalysts dispersed on Si(100) the mass uptake is linear throughout the process. On high surface area catalysts, however, a saturation of the mass uptake is indifferently observed. This saturation is explained either by diffusion limitation by the growing MWCNTs or by internal diffusion through the pores or external diffusion through the grains of the catalyst. The kinetic dependence with partial pressure of the incoming C2H6:H2 gas mixture is then explored on the FeAl2O3 catalyst. A linear dependence of the MWCNT growth an (P(C2H6)/P(H2))(1/2) is found. A simple model is then developed that accounts for this dependence only if an associative and competitive adsorption of ethane is the rate determining step of the overall process. These results thus bring insight to improve and control the CCVD growth kinetics of MWCNTs. PMID- 16709130 TI - Anisotropic optical properties in electroluminescent conjugated polymers based on grazing angle photoluminescence measurements. AB - Grazing angle photoluminescence (GPL) originates from a waveguided light emitted at grazing angle to the substrate due to the total internal reflections, and the light emission is polarized with enhanced intensity at selective mode wavelength. GPL measurements reveal the optical anisotropy of luminescent conjugated polymers, in particular, the alignment of emitting dipoles from which emission occurs, in contrast to spectroscopic ellipsometry measurements that give the anisotropy in the absorption. Based on the GPL emission intensities and spectra, we investigate the anisotropic optical properties in electroluminescent poly(9,9' di-n-octylfluorene-alt-benzothiadiazole) (F8BT) conjugated polymer thin films of different molecular weights (M(n) = 9-255 kg/mol), both in the pristine and annealed states. The optical anisotropy in F8BT films generally increases with molecular weight, suggesting that higher molecular weight polymers with longer chains are more likely to lie in-plane to the substrate. Upon annealing, high molecular weight F8BT films show even a higher degree of anisotropy, in contrast to low molecular weight F8BT films that become more isotropic. Annealing causes the polymer chains to rearrange and adopt a configuration in which the interchain exciton migration to better ordered low energy (LE) emissive states is strongly suppressed. We observe that the emissive states in F8BT are strongly affected by the local polymer chain arrangement, producing the less ordered high energy (HE) emissive states near the substrate interface where there is a higher degree of chain disorder and the LE states in the bulk of the film. When spin coated onto a quartz substrate precoated with a poly(styrenesulfonate)-doped poly(3,4 ethylenedioxythiophene) (PEDOT:PSS) layer, films of F8BT show severe luminescence quenching near the PEDOT:PSS interface for both the LE and HE emissive states, but a selective quenching of the LE states in the bulk of the film. These observations have important implications for fabricating efficient electronic devices using conjugated polymers as an active material, since the performance of these devices will strongly depend on anisotropic optical properties of electroluminescent conjugated polymers. PMID- 16709131 TI - Evolution of electronic structure and spectral evaluation in single-crystal Mn3O4 nanorods. AB - Single-crystal Mn3O4 nanorods with tetragonal structure have been successfully prepared by a chemical reaction route. Transmission electron microscopy (TEM) and high-resolution TEM studies prove that the single-crystal Mn3O4 nanorod is smooth and straight, and that the geometrical shape is structurally perfect. We investigated the electronic characteristics of Mn3O4 nanorods by various spectral evaluations. The present study confirms that the hybridization between oxygen 2p and manganese 3d orbits plays an important role when considering electronic structures of Mn3O4 nanorods. PMID- 16709132 TI - Simulated annealing study of asymmetric diblock copolymer thin films. AB - We report a simulated annealing study of the morphology of asymmetric diblock copolymer thin films confined between two homogeneous and identical surfaces. We have focused on copolymers that form a gyroidal morphology in the bulk. The morphological dependence of the confined films on the film thickness and the surface-polymer interaction has been systematically investigated. From the simulations it is found that much richer morphologies can form for the gyroid forming asymmetric diblock copolymer thin films, in contrast to the lamella forming symmetric and cylinder-forming asymmetric diblock copolymer films. Multiple morphological transitions induced by changing the film thickness and polymer-surface interactions are observed. PMID- 16709133 TI - A molecular dynamics study of free energy of micelle formation for sodium dodecyl sulfate in water and its size distribution. AB - Free energy of micelle formation has been evaluated for spherical sodium dodecyl sulfate (SDS) in water by a thermodynamic integration method combined with a series of large-scale molecular dynamics calculations following the chemical species model. In particular, free energy change delta mu(n+1)0 with respect to the addition of one surfactant molecule to the spherical micelle of size n was obtained as a function of n. The free energy profile showed a minimum followed by a maximum, which corresponds to a peak in the size distribution. The calculated peak size n = 57 near its critical micelle concentration is in good agreement with the experimental averaged aggregation number n = 55-75 of the SDS micelle. The distribution showed a rather sharp peak, indicating that the size is almost a monodisperse one. The size is likely to be insensitive to the total concentration of the surfactant. PMID- 16709134 TI - Molecular dynamics simulation of polymer electrolytes based on poly(ethylene oxide) and ionic liquids. I. Structural properties. AB - Molecular dynamics (MD) simulations have been performed for prototype models of polymer electrolytes in which the salt is an ionic liquid based on 1-alkyl-3 methylimidazolium cations and the polymer is poly(ethylene oxide), PEO. The MD simulations were performed by combining the previously proposed models for pure ionic liquids and polymer electrolytes containing simple inorganic ions. A systematic investigation of ionic liquid concentration, temperature, and the 1 alkyl- chain length, [1,3-dimethylimidazolium]PF6, and [1-butyl-3 methylimidazolium]PF6, effects on resulting equilibrium structure is provided. It is shown that the ionic liquid is dispersed in the polymeric matrix, but ionic pairs remain in the polymer electrolyte. Imidazolium cations are coordinated by both the anions and the oxygen atoms of PEO chains. Probability density maps of occurrences of nearest neighbors around imidazolium cations give a detailed physical picture of the environment experienced by cations. Conformational changes on PEO chains upon addition of the ionic liquid are identified. The equilibrium structure of simulated systems is also analyzed in reciprocal space by using the static structure factor, S(k). Calculated S(k) display a low wave vector peak, indicating that spatial correlation in an extended-range order prevail in the ionic liquid polymer electrolytes. Long-range correlations are assigned to nonuniform distribution of ionic species within the simulation box. PMID- 16709135 TI - A coarse-grained model for the formation of alpha helix with a noninteger period on simple cubic lattices. AB - Periodicity is an important parameter in the characterization of a helix in proteins. In this work, a coarse-grained model for a homopolypeptide in simple cubic lattices has been extended to build an alpha helix with a noninteger period. The lattice model is based on the bond fluctuation algorithm in which bond lengths and orientations are altered in a quasicontinuous way, and the simulation is performed via dynamic Monte Carlo simulation. Hydrogen bonds are assumed to be formed between a virtual-carbonyl group in a residue and a downstream virtual-imino group in another residue. Consequently, the period of the formed alpha helix is a noninteger. An improved spatial correlation function has been suggested to quantitatively describe the periodicity of the helical conformation, by which helical period and helical persistent length can be calculated by statistics. The resultant periods are very close to 3.6 residues; the persistent length based upon the improved definition can be larger or smaller than the chain length and reflect the inherent regularity of a chain including one or multiple helical blocks. The simulation outputs agree with the calculation of the Zimm-Bragg theory based upon the associated nucleation parameter and propagation parameter as well. PMID- 16709136 TI - Simple model of membrane proteins including solvent. AB - We report a numerical simulation for the phase diagram of a simple two dimensional model, similar to the one proposed by Noro and Frenkel [J. Chem. Phys. 114, 2477 (2001)] for membrane proteins, but one that includes the role of the solvent. We first use Gibbs ensemble Monte Carlo simulations to determine the phase behavior of particles interacting via a square-well potential in two dimensions for various values of the interaction range. A phenomenological model for the solute-solvent interactions is then studied to understand how the fluid fluid coexistence curve is modified by solute-solvent interactions. It is shown that such a model can yield systems with liquid-liquid phase separation curves that have both upper and lower critical points, as well as closed loop phase diagrams, as is the case with the corresponding three-dimensional model. PMID- 16709137 TI - Equilibrium conformational dynamics of a polymer in a solvent. AB - Molecular dynamics simulations were used to study the conformational dynamics of a bead-spring model polymer in an explicit solvent under good solvent conditions. The dynamics of the polymer chain were investigated using an analysis of the time autocorrelation functions of the Rouse coordinates of the polymer chain. We have investigated the variation of the correlation functions with polymer chain length N, solvent density rho, and system size. The measured initial decay rates gamma(p) of the correlation functions were compared with the predictions from a theory of polymer dynamics which uses the Oseen tensor to describe hydrodynamic interactions between monomers. Over the range of chain lengths considered (N = 30 60 monomers), the predicted scaling of gamma(p) proportional to N(-3nu) was observed at high rho, where nu is the polymer scaling exponent. The predicted gamma(p) are generally higher than the measured values. This discrepancy increases with decreasing rho, as a result in the breakdown in the conditions required for the Oseen approximation. The agreement between theory and simulation at high rho improves considerably if the theoretical expression for gamma(p) is modified to avoid sum-to-integral approximations, and if the values of (R(p)2), which are used in the theory, are taken directly from the simulation rather than being calculated using approximate scaling relations. The observed finite-size scaling of gamma(p) is not quantitatively consistent with the theoretical predictions. PMID- 16709138 TI - Is there a higher-order mode coupling transition in polymer blends? AB - We present simulations on a binary blend of bead-spring polymer chains. The introduction of monomer size disparity yields very different relaxation times for each component of the blend. Competition between two different arrest mechanisms, namely, bulklike dynamics and confinement, leads to an anomalous relaxation scenario for the fast component, characterized by sublinear time dependence for mean squared displacements, or logarithmic decay and convex-to-concave crossover for density-density correlators. These anomalous dynamic features, which are observed over time intervals extending up to 4 decades, strongly resemble predictions of mode coupling theory for nearby higher-order transitions. Chain connectivity extends anomalous relaxation over a wide range of blend compositions. PMID- 16709139 TI - Comment on "On the dielectric susceptibility spectra of supercooled o-terphenyl" [J. Chem. Phys. 123, 154502 (2005)]. PMID- 16709143 TI - Ultraviolet B light stimulates interleukin-20 expression by human epithelial keratinocytes. AB - The proinflammatory cytokine interleukin-20 (IL-20) may exert the majority of its activity in the skin. We examined the effect of various treatments including several forms of phototherapy on IL-20 expression using cultured normal human epithelial keratinocytes (NHEK). Broadband UVB light, recombinant (r) IL-1 and rIL-8 increased, while hydrocortisone reduced, NHEK supernatant IL-20 levels. Elevation of NHEK IL-20 mRNA and maximal supernatant IL-20 levels occurred with a UVB light dose (40 mJ cm(-2)) that reduced cell viability by approximately 50%. While this UVB light dose also elevated supernatant IL-1 alpha and IL-8 levels, antibody neutralization studies indicated that neither of these cytokines was directly responsible for this increase in IL-20 expression. However, the elevation in IL-20 levels was fully inhibited by the p38 mitogen-activated protein kinase (MAPK) inhibitor SB-203580, suggesting involvement of this stress signaling pathway in this UVB light response. Photodynamic therapy (PDT) with the photosensitizer lemuteporfin, UVA light, cisplatin, lipopolysaccharide (LPS), tumor necrosis factor-alpha (TNF-alpha) or recombinant interferon-gamma (rIFN gamma) either had little effect or decreased NHEK supernatant IL-20 levels. Reduced IL-20 levels paralleled the cytotoxic actions of PDT, UVA light or cisplatin and the antiproliferative effect of rIFN-gamma. Neither rIL-20 supplementation nor anti-IL-20 antibody treatments affected cell viability indicating that soluble IL-20 did not affect the short-term survival of UVB light irradiated NHEK. Stimulation of IL-20 expression in keratinocytes by UVB light suggests that this cytokine might participate in skin responses to this ever present environmental factor and potentially has a role in UV light-associated dermatoses. PMID- 16709144 TI - Analogue chromophore study of the influence of electronic perturbation on color regulation of photoactive yellow protein. AB - We report a unique lambdamax shift of the absorption maximum of a photoactive yellow protein (PYP) analogue reconstituted with a fluorinated chromophore (F PYP). The difference in lambdamax between the free chromophore and the protein was significantly larger than that with the native chromophore. We concluded that the unusual lambdamax shift is caused by the electronegative character of the fluorine atom and not by steric hindrance. This result suggests that formation of a hydrogen bond between the fluorine atom and one or more amino acid residues could neutralize its electron-withdrawing character. The properties of analogues of PYP with brominated and methylated chromophore could be explained as an effect of steric hindrance. PMID- 16709145 TI - New strategies of photoprotection. AB - Adequate photoprotection is essential to control UV-related disorders, including sunburn, photoaging and photocarcinogenisis. Sun avoidance, protection of skin with clothing, and sunscreens are presently the best way of photoprotection, assuming that they are used properly. However, new strategies, which are based on or make use of the endogenous protective response to UV light, may further improve currently used photoprotective means. The addition of repair enzymes and/or antioxidants has a positive effect on skin's recovery from UV-induced DNA damage. Several botanical agents, mainly vitamins and polyphenols, have shown to influence signal transduction pathways leading to photoprotective effects. Also stimulation of endogenous UV-response pathways via irradiation with a low UV dose or via simulation of UV-induced DNA-damage results in photoprotective effects. Future research in this field and combination of different photoprotective strategies will hopefully lead to improved photoprotection. PMID- 16709150 TI - Agrobacterium tumefaciens and plant cell interactions and activities required for interkingdom macromolecular transfer. AB - Host recognition and macromolecular transfer of virulence-mediating effectors represent critical steps in the successful transformation of plant cells by Agrobacterium tumefaciens. This review focuses on bacterial and plant-encoded components that interact to mediate these two processes. First, we examine the means by which Agrobacterium recognizes the host, via both diffusible plant derived chemicals and cell-cell contact, with emphasis on the mechanisms by which multiple host signals are recognized and activate the virulence process. Second, we characterize the recognition and transfer of protein and protein-DNA complexes through the bacterial and plant cell membrane and wall barriers, emphasizing the central role of a type IV secretion system-the VirB complex-in this process. PMID- 16709151 TI - Saccharomyces cerevisiae cells have three Omega class glutathione S-transferases acting as 1-Cys thiol transferases. AB - The Saccharomyces cerevisiae genome encodes three proteins that display similarities with human GSTOs (Omega class glutathione S-transferases) hGSTO1-1 and hGSTO2-2. The three yeast proteins have been named Gto1, Gto2 and Gto3, and their purified recombinant forms are active as thiol transferases (glutaredoxins) against HED (beta-hydroxyethyl disulphide), as dehydroascorbate reductases and as dimethylarsinic acid reductases, while they are not active against the standard GST substrate CDNB (1-chloro-2,4-dinitrobenzene). Their glutaredoxin activity is also detectable in yeast cell extracts. The enzyme activity characteristics of the Gto proteins contrast with those of another yeast GST, Gtt1. The latter is active against CDNB and also displays glutathione peroxidase activity against organic hydroperoxides such as cumene hydroperoxide, but is not active as a thiol transferase. Analysis of point mutants derived from wild-type Gto2 indicates that, among the three cysteine residues of the molecule, only the residue at position 46 is required for the glutaredoxin activity. This indicates that the thiol transferase acts through a monothiol mechanism. Replacing the active site of the yeast monothiol glutaredoxin Grx5 with the proposed Gto2 active site containing Cys46 allows Grx5 to retain some activity against HED. Therefore the residues adjacent to the respective active cysteine residues in Gto2 and Grx5 are important determinants for the thiol transferase activity against small disulphide-containing molecules. PMID- 16709152 TI - Direct comparison of relaxation and cGMP production in human coronary by-pass grafts in response to stimulation with natriuretic peptides and a nitric oxide donor. AB - In the present study, we investigated the vasodilator properties of A-type, B type and C-type natriuretic peptides (ANP, BNP and CNP respectively) and the NO (nitric oxide) donor sin-1 (3-morpholino-sydnonimine) in human by-pass grafts. In contrast with previous studies, the same vessel was used to demonstrate a direct link between cGMP production and functional relaxation. Remnants of the IMA (internal mammary artery) and SV (saphenous vein) were obtained from 82 patients undergoing coronary artery by-pass grafting. The responses to cumulative concentrations of ANP, BNP, CNP and sin-1 in vessel rings pre-contracted with a thromboxane A2 agonist (U46619) were measured in an organ bath. Additionally, intracellular cGMP production after single submaximal dose application of these drugs to vessel rings was determined by a RIA. ANP (P=0.001) and sin-1 (P<0.001) caused significant concentration-dependent relaxation of the IMA. In the SV, only sin-1 (P<0.001) induced marked concentration-dependent relaxation. At a single submaximal concentration, significant relaxation as well as intracellular cGMP production were found in response to ANP, BNP and sin-1 in the IMA. In contrast, in the SV, only sin-1 significantly induced cGMP production and relaxation. There was a moderate, but significant, correlation between intracellular cGMP net production and net relaxation in the IMA. In conclusion, ANP, as the most powerful relaxant of all the natriuretic peptides tested on the IMA, may be a possible alternative vasorelaxant to overcome peri-operative vasospasm in this artery. In contrast with sin-1, ANP and BNP were not effective vasorelaxants of the SV. Net relaxation in response to natriuretic peptides correlated with cGMP net concentrations in the IMA. PMID- 16709154 TI - WITHDRAWN: Constitutive expression and characterization of Hepatitis B surface antigen purified by metal affinity precipitation. AB - The paper entitled "Constitutive expression and characterization of Hepatitis B surface antigen purified by metal affinity precipitation", which was published online on 19 May 2006, was withdrawn at the author's request. PMID- 16709155 TI - Sterol and lipid trafficking in mammalian cells. AB - The pathways involved in the intracellular transport and distribution of lipids in general, and sterols in particular, are poorly understood. Cholesterol plays a major role in modulating membrane bilayer structure and important cellular functions, including signal transduction and membrane trafficking. Both the overall cholesterol content of a cell, as well as its distribution in specific organellar membranes are stringently regulated. Several diseases, many of which are incurable at present, have been characterized as results of impaired cholesterol transport and/or storage in the cells. Despite their importance, many fundamental aspects of intracellular sterol transport and distribution are not well understood. For instance, the relative roles of vesicular and non-vesicular transport of cholesterol have not yet been fully determined, nor are the non vesicular transport mechanisms well characterized. Similarly, whether cholesterol is asymmetrically distributed between the two leaflets of biological membranes, and if so, how this asymmetry is maintained, is poorly understood. In this review, we present a summary of the current understanding of these aspects of intracellular trafficking and distribution of lipids, and more specifically, of sterols. PMID- 16709153 TI - Ras activation in response to phorbol ester proceeds independently of the EGFR via an unconventional nucleotide-exchange factor system in COS-7 cells. AB - Ras is a major mediator of PE (phorbol ester) effects in mammalian cells. Various mechanisms for PE activation of Ras have been reported [Downward, Graves, Warne, Rayter and Cantrell (1990) Nature (London) 346, 719-723; Shu, Wu, Mosteller and Broek (2002) Mol. Cell. Biol. 22, 7758-7768; Roose, Mollenauer, Gupta, Stone and Weiss (2005) Mol. Cell. Biol. 25, 4426-4441; Grosse, Roelle, Herrlich, Hohn and Gudermann (2000) J. Biol. Chem. 275, 12251-12260], including pathways that target GAPs (GTPase-activating proteins) for inactivation and those that result in activation of GEFs (guanine nucleotide-exchange factors) Sos (son of sevenless homologue) or RasGRP (RAS guanyl releasing protein). However, a biochemical link between PE and GAP inactivation is missing and GEF stimulation is hard to reconcile with the observation that dominant-negative S17N-Ras does not compromise Ras-dependent ERK (extracellular-signal-regulated kinase) activation by PE. We have addressed this controversy and carried out an in-depth biochemical study of PE-induced Ras activation in COS-7 cells. Using a cell-permeabilization approach to monitor nucleotide exchange on Ras, we demonstrate that PE-induced Ras-GTP accumulation results from GEF stimulation. Nucleotide exchange stimulation by PE is prevented by PKC (protein kinase C) inhibition but not by EGFR [EGF (epidermal growth factor) receptor] blockade, despite the fact that EGFR inhibition aborts basal and PE-induced Shc (Src homology and collagen homology) phosphorylation and Shc-Grb2 (growth-factor-receptor-bound protein 2) association. In fact, EGFR inhibition ablates basal nucleotide exchange on Ras in growth-arrested COS-7 cells. These data disclose the existence of two separate GEF systems that operate independently from each other to accomplish PE-dependent formation of Ras-GTP and to maintain resting Ras-GTP levels respectively. We document that COS-7 cells do not express RasGRP and present evidence that the PE responsive GEF system may involve PKC-dependent phosphorylation of Sos. More fundamentally, these observations shed new light on enigmatic issues such as the inefficacy of S17N-Ras in blocking PE action or the role of the EGFR in heterologous agonist activation of the Ras/ERK pathway. PMID- 16709156 TI - Nucleus-vacuole junctions in yeast: anatomy of a membrane contact site. AB - NV junctions (nucleus-vacuole junctions) in Saccharomyces cerevisiae are MCSs (membrane contact sites) formed through specific interactions between Vac8p on the vacuole membrane and Nvj1p in the outer nuclear membrane, which is continuous with the perinuclear ER (endoplasmic reticulum). NV junctions mediate a unique autophagic process that degrades portions of the yeast nucleus through a process called 'piecemeal microautophagy of the nucleus' (PMN). Our studies suggest that the lipid composition of NV junctions plays an important role in the biogenesis of PMN structures. NV junctions represent a unique model system for studying the biology of ER MCSs, as well as the molecular mechanism of selective microautophagy. PMID- 16709157 TI - MLN64 and MENTHO, two mediators of endosomal cholesterol transport. AB - MLN64 (metastatic lymph node 64) and MENTHO (MLN64 N-terminal homologue) are two late-endosomal proteins that share a conserved region of four transmembrane helices with three short intervening loops called the MENTAL domain (MLN64 N terminal domain). This domain mediates MLN64 and MENTHO homo- and hetero interactions, targets both proteins to late endosomes and binds cholesterol in vivo. In addition to the MENTAL domain, MLN64 contains a cholesterol-specific START domain [StAR (steroidogenic acute regulatory protein)-related lipid transfer domain]. The START domain is a protein module of approx. 210 residues that binds lipids, including sterols, and is present in 15 distinct proteins in mammals. Thus MLN64 and MENTHO define discrete cholesterol-containing subdomains within the membrane of late endosomes where they may function in cholesterol transport. The MENTAL domain might serve to maintain cholesterol at the membrane of late endosomes prior to its shuttle to cytoplasmic acceptor(s) through the START domain. PMID- 16709158 TI - Sec14p-like proteins regulate phosphoinositide homoeostasis and intracellular protein and lipid trafficking in yeast. AB - The major PI (phosphatidylinositol)/PC (phosphatidylcholine)-transfer protein in yeast, Sec14p, co-ordinates lipid metabolism with protein transport from the Golgi complex. Yeast also express five additional gene products that share 24-65% primary sequence identity with Sec14p. These Sec14p-like proteins are termed SFH (Sec Fourteen Homologue) proteins, and overexpression of certain individual SFH gene products rescues sec14-1(ts)-associated growth and secretory defects. SFH proteins are atypical in that these stimulate the transfer of PI, but not PC, between distinct membrane bilayer systems in vitro. Further analysis reveals that SFH proteins functionally interact with the Stt4p phosphoinositide 4-kinase to stimulate PtdIns(4,5)P(2) synthesis which in turn activates phospholipase D. Finally, genetic analyses indicate that Sfh5p interfaces with the function of specific subunits of the exocyst complex as well as the yeast SNAP-25 (25 kDa synaptosome-associated protein) homologue, Sec9p. Our current view is that Sfh5p regulates PtdIns(4,5)P(2) homoeostasis at the plasma membrane, and that Sec9p responds to that regulation. Thus SFH proteins individually regulate specific aspects of lipid metabolism that couple, with exquisite specificity, with key cellular functions. PMID- 16709159 TI - Moving Ca2+ from the endoplasmic reticulum to mitochondria: is spatial intimacy enough? AB - A number of studies in recent years have demonstrated that the ER (endoplasmic reticulum) makes intimate contacts with mitochondria, the latter organelles existing both as individual organelles and occasionally as a more extensive interconnected network. Demonstrations that mitochondria take up Ca(2+) more avidly upon its mobilization from the ER than when delivered to permeabilized cells as a buffered solution also indicate that a shielded conduit for Ca(2+) may exist between the two organelle types, perhaps comprising the inositol 1,4,5 trisphosphate receptor and mitochondrial outer membrane proteins including the VDAC (voltage-dependent anion channel). Although the existence of such intracellular ER-mitochondria 'synapses', or of an ER-mitochondria Ca(2+) 'translocon', is an exciting idea, more definitive experiments are needed to test this possibility. PMID- 16709160 TI - Sterol trafficking between the endoplasmic reticulum and plasma membrane in yeast. AB - We recently showed that transport of ergosterol from the ER (endoplasmic reticulum) to the sterol-enriched PM (plasma membrane) in yeast occurs by a non vesicular (Sec18p-independent) mechanism that results in the equilibration of sterol pools in the two organelles [Baumann, Sullivan, Ohvo-Rekila, Simonot, Pottekat, Klaassen, Beh and Menon (2005) Biochemistry 44, 5816-5826]. To explore how this occurs, we tested the role of proteins that might act as sterol transporters. We chose to study oxysterol-binding protein homologues (Osh proteins), a family of seven proteins in yeast, all of which contain a putative sterol-binding pocket. Recent structural analyses of one of the Osh proteins [Im, Raychaudhuri, Prinz and Hurley (2005) Nature (London) 437, 154-158] suggested a possible transport cycle in which Osh proteins could act to equilibrate ER and PM pools of sterol. Our results indicate that the transport of newly synthesized ergosterol from the ER to the PM in an OSH deletion mutant lacking all seven Osh proteins is slowed only 5-fold relative to the isogenic wild-type strain. Our results suggest that the Osh proteins are not sterol transporters themselves, but affect sterol transport in vivo indirectly by affecting the ability of the PM to sequester sterols. PMID- 16709161 TI - Uptake and trafficking of exogenous sterols in Saccharomyces cerevisiae. AB - The proper distribution of sterols among organelles is critical for numerous cellular functions. How sterols are sorted and moved among membranes remains poorly understood, but they are transported not only in vesicles but also by non vesicular pathways. One of these pathways moves exogenous sterols from the plasma membrane to the endoplasmic reticulum in the yeast Saccharomyces cerevisiae. We have found that two classes of proteins play critical roles in this transport, ABC transporters (ATP-binding-cassette transporters) and oxysterol-binding protein-related proteins. Transport is also regulated by phosphoinositides and the interactions of sterols with other lipids. Here, we summarize these findings and speculate on the role of non-vesicular sterol transfer in determining intracellular sterol distribution and membrane function. PMID- 16709162 TI - Lipid homoeostasis and Golgi secretory function. AB - The unique lipid composition of the Golgi membranes is critical for maintaining their structural and functional identity, and is regulated by local lipid metabolism, a variety of lipid-binding, -modifying, -sensing and -transfer proteins, and by selective lipid sorting mechanisms. A growing body of evidence suggests that certain lipids, such as phosphoinositides and diacylglycerol, regulate Golgi-mediated transport events. However, their exact role in this process, and the underlying mechanisms that maintain their critical levels in specific membrane domains of the Golgi apparatus, remain poorly understood. Nevertheless, recent advances have revealed key regulators of lipid homoeostasis in the Golgi complex and have demonstrated their role in Golgi secretory function. PMID- 16709163 TI - Organization and functions of sphingolipid biosynthesis in yeast. AB - Sphingolipids are important signalling molecules and thus it is very important to understand how they are generated. Sphingolipid biosynthesis shows a conserved compartmentalization in eukaryotic cells. Their synthesis begins in the endoplasmic reticulum and is completed in the Golgi apparatus. We now know quite a bit about the topology of the reactions in the pathway, but certain critical steps, such as ceramide synthesis, are still poorly understood. In the present paper, we discuss the latest views on this subject. PMID- 16709164 TI - Interaction of the smooth endoplasmic reticulum and mitochondria. AB - The ER (endoplasmic reticulum) is composed of multiple domains including the nuclear envelope, ribosome-studded rough ER and the SER (smooth ER). The SER can also be functionally segregated into domains that regulate ER-Golgi traffic (transitional ER), ERAD (ER-associated degradation), sterol and lipid biosynthesis and calcium sequestration. The last two, as well as apoptosis, are critically regulated by the close association of the SER with mitochondria. Studies with AMFR (autocrine motility factor receptor) have defined an SER domain whose integrity and mitochondrial association can be modulated by ilimaquinone as well as by free cytosolic calcium levels in the normal physiological range. AMFR is an E3 ubiquitin ligase that targets its ligand directly to the SER via a caveolae/raft-dependent pathway. In the present review, we will address the relationship between the calcium-dependent morphology and mitochondrial association of the SER and its various functional roles in the cell. PMID- 16709165 TI - Regulation of ecto-5'-nucleotidase activity via Ca2+-dependent, annexin 2 mediated membrane rearrangement? AB - The spatial segregation of the plasma membrane plays a prominent role in distinguishing and sorting a large number of signals a cell receives simultaneously. The plasma membrane comprises regions known as lipid rafts, which serve as signal-transduction hubs and platforms for sorting membrane-associated proteins. Ca(2+)-binding proteins of the annexin family have been ascribed a role in the regulation of raft dynamics. Glycosylphosphatidylinositol-anchored 5' nucleotidase is an extracellular, raft-associated enzyme responsible for conversion of extracellular ATP into adenosine. Our results point to a regulation of ecto-5'-nucleotidase activity by Ca(2+)-dependent, annexin-mediated stabilization of membrane rafts. PMID- 16709166 TI - Phosphatidylinositol-transfer protein and its homologues in yeast. AB - Yeast Sec14p acts as a phosphatidylinositol/phosphatidylcholine-transfer protein in vitro. In vivo, it is essential in promoting Golgi secretory function. Products of five genes named SFH1-SFH5 (Sec Fourteen Homologues 1-5) exhibit significant sequence homology to Sec14p and together they form the Sec14p family of lipid-transfer proteins. It is a diverse group of proteins with distinct subcellular localizations and varied physiological functions related to lipid metabolism and membrane trafficking. PMID- 16709167 TI - Movement of calcium signals and calcium-binding proteins: firewalls, traps and tunnels. AB - In the board game 'Snakes and Ladders', placed on the image of a pancreatic acinar cell, calcium ions have to move from release sites in the secretory region to the nucleus. There is another important contraflow - from calcium entry channels in the basal part of the cell to ER (endoplasmic reticulum) terminals in the secretory granule region. Both transport routes are perilous as the messenger can disappear in any place on the game board. It can be grabbed by calcium ATPases of the ER (masquerading as a snake but functioning like a ladder) and tunnelled through its low buffering environment, it can be lured into the whirlpools of mitochondria uniporters and forced to regulate the tricarboxylic acid cycle, and it can be permanently placed inside the matrix of secretory granules and released only outside the cell. The organelles could trade calcium (e.g. from the ER to mitochondria and vice versa) almost depriving this ion the light of the cytosol and noble company of cytosolic calcium buffers. Altogether it is a rich and colourful story. PMID- 16709168 TI - Endoplasmic reticulum generates calcium signalling microdomains around the nucleus and spindle in syncytial Drosophila embryos. AB - Cell cycle calcium signals are generated by inositol trisphosphate-mediated release of calcium from internal stores [Ciapa, Pesando, Wilding and Whitaker (1994) Nature (London) 368, 875-878; Groigno and Whitaker (1998) Cell 92, 193 204]. The major internal calcium store is the ER (endoplasmic reticulum): the spatial organization of the ER during mitosis is important in defining a microdomain around the nucleus and mitotic spindle in early Drosophila embryos [Parry, McDougall and Whitaker (2005) J. Cell Biol. 171, 47-59]. Nuclear divisions in syncytial Drosophila embryos are accompanied by both cortical and nuclear localized calcium transients. Mitosis is prevented by the InsP(3) antagonists Xestospongin C and heparin. Nuclear-localized transients and cortical transients rely on extraembryonic calcium, suggesting that ER calcium levels are maintained by calcium influx. PMID- 16709169 TI - The OSBP-related proteins (ORPs): global sterol sensors for co-ordination of cellular lipid metabolism, membrane trafficking and signalling processes? AB - Protein families related to OSBP (oxysterol-binding protein) are present in eukaryotes from yeast to human. The functions of the ORPs (OSBP-related proteins) have remained largely enigmatic. Even though they have been implicated in the function of ERJs (endoplasmic reticulum junctions), it is evident that any single model for their mechanism of action is insufficient. The existing evidence points in many different directions, such as integration of sterol and sphingomyelin metabolism, regulation of neutral lipid metabolism, control of signalling cascades, regulation of secretory vesicle generation, and function in the microtubule-based motility of endo/lysosomes. Some of these functions could involve ERJ and non-vesicular transport of lipids, but this is unlikely to be the unifying feature. We believe, rather, that the common denominator for ORP function is acting as sterol sensors that relay information to a spectrum of cellular processes. PMID- 16709170 TI - Endosomal cholesterol traffic: vesicular and non-vesicular mechanisms meet. AB - The endoplasmic reticulum is traditionally perceived as the key compartment for regulating intracellular cholesterol metabolism. Increasing evidence suggests that the endocytic pathway provides an additional regulatory level governing intracellular cholesterol trafficking and homoeostasis. Sterols can enter, and apparently also exit, endosomal compartments via both vesicular and non-vesicular mechanisms. A number of studies have focused on endosomal sterol removal as its defects lead to cholesterol storage diseases. So far, the bulk of evidence on endosomal sterol egress describes the involvement of membrane trafficking machineries. Interestingly, two late endosomal sterol-binding proteins were recently shown to regulate the movement of late endosomes along cytoskeletal tracks. These studies provide the first indications of how non-vesicular and vesicular mechanisms may co-operate in endosomal sterol trafficking. PMID- 16709171 TI - Lipid trafficking between the endoplasmic reticulum and the chloroplast. AB - The photosynthetic (thylakoid) membrane of plants is one of the most extensive biological cell membrane systems found in Nature. It harbours the photosynthetic apparatus, which is essential to life on Earth as carbon dioxide is fixed and atmospheric oxygen released by photosynthesis. Lipid biosynthetic enzymes of different subcellular compartments participate in the biogenesis of the thylakoid membrane system. This process requires the extensive exchange of lipid precursors between the chloroplast and the ER (endoplasmic reticulum). The underlying lipid trafficking phenomena are not yet understood at the mechanistic level, but genetic mutants of the model plant Arabidopsis thaliana with disruptions in lipid trafficking between the ER and the chloroplast have recently become available. Their study has led to the identification of components of the lipid transfer machinery at the inner chloroplast envelope. PMID- 16709172 TI - Lipid dynamics in neurons. AB - Compared with other organs, the brain is highly enriched in cholesterol. Essentially all cholesterol in the brain is synthesized within the brain; the blood-brain barrier prevents the import of plasma lipoproteins into the brain. Consequently, the brain operates an independent lipoprotein transport system in which glial cells produce ApoE (apolipoprotein E)-containing lipoproteins that are thought to deliver cholesterol to neurons for axonal growth and repair. We have shown that ApoE-containing lipoproteins generated by glial cells stimulate axon extension. ApoE associated with lipoprotein particles, and a receptor of the low-density lipoprotein receptor family, are required for stimulation of axon growth. NPC (Niemann-Pick type C) disease is a severe neurological disorder caused by mutations in the NPC1 or NPC2 gene. A hallmark of this disease is impaired transport of cholesterol out of late endosomes/lysosomes and the accumulation of cholesterol in these organelles. Although cholesterol accumulates in cell bodies of neurons from NPC1-deficient mice, the cholesterol content of axons is reduced. The presence of NPC1 in endosomal structures in nerve terminals, and the finding of aberrant synaptic vesicles, suggest that defects in synaptic vesicle recycling contribute to neurological abnormalities characteristic of NPC disease. We have also shown that ApoE-containing lipoproteins produced by glial cells from NCP1-deficient mice are of normal composition and stimulate axon extension. PMID- 16709174 TI - Cytokines and proteoglycans: an introductory overview. AB - The defining characteristic of the glycoproteins known as proteoglycans is the presence of O-linked acidic polysaccharides known as GAGs (glycosaminoglycans). The backbone of these linear polysaccharides is a repeating disaccharide, comprising N-acetyl hexosamine alternating with beta-D-glucuronic acid, alpha-L iduronic acid, or galactose. For some GAGs, partial deacetylation, epimerization of glucuronic acid, and substitution with N- and O-sulphates result in highly complex, heterogeneous structures. The interactions with proteins through which GAGs exert their biological effects depend on the resulting sequences. Some proteins, for example antithrombin, have highly specific sequence requirements for their GAG ligand [in this case heparin or HS (heparan sulphate)]; others, for example the fibroblast growth factors, are less demanding. GAGs, in particular HS, play a role as co-receptors for some cytokines. In addition, HS is thought to be important for the localization of cytokines, acting both as a tissue store and as a mediator of morphogen gradient formation in development. The structural determinants of GAG-cytokine interactions are therefore clearly important to understanding the biology of development, wound healing and the immune system. No single paradigm has been identified for such interactions, and the search for general principles underlying involvement of GAGs in cytokine function is at an early stage. PMID- 16709173 TI - Macromolecular assemblies regulate nonvesicular phosphatidylserine traffic in yeast. AB - PtdSer (phosphatidylserine) is synthesized in the endoplasmic reticulum and the related MAM (mitochondria-associated membrane), and transported to the PtdSer decarboxylases, Pds1p in the mitochondria, and Psd2p in the Golgi. Genetic and biochemical analyses of PtdSer transport are now revealing the role of specific protein and lipid assemblies on different organelles that regulate non-vesicular PtdSer transport. The transport of PtdSer from MAM to mitochondria is regulated by at least three genes: MET30 (encoding a ubiquitin ligase), MET4 (encoding a transcription factor), and one or more unknown genes whose transcription is regulated by MET4. MET30-dependent ubiquitination is required for the MAM to function as a competent donor membrane and for the mitochondria to function as a competent acceptor membrane. Non-vesicular transport of PtdSer to the locus of Psd2p is under the control of at least three genes, STT4 [encoding Stt4p (phosphatidylinositol 4-kinase)], PSTB2 (encoding the lipid-binding protein PstB2p) and PSD2 (encoding Psd2p). Stt4p is proposed to produce a pool of PtdIns4P that is necessary for lipid transport. PstB2p and Psd2p must be present on the acceptor membrane for PtdSer transport to occur. Psd2p contains a C2 (Ca(2+) and phospholipid binding sequence) domain that is required for lipid transport. Reconstitution studies with chemically defined donor membranes demonstrate that membrane domains rich in the anionic lipids, PtdSer, PtdIns4P and phosphatidic acid function as the most efficient donors of PtdSer to Psd2p. The emerging view is that macromolecular complexes dependent on protein-protein and protein-lipid interactions form between donor and acceptor membranes and serve to dock the compartments and facilitate phospholipid transport. PMID- 16709175 TI - Signalling by HGF/SF and Met: the role of heparan sulphate co-receptors. AB - The receptor tyrosine kinase Met and its ligand HGF/SF (hepatocyte growth factor/scatter factor) are essential in the signalling pathways required for embryogenesis and tissue regeneration. Aberrant signalling of this complex is also a feature of many tumours and appears to contribute to the growth, invasiveness and metastasis of both carcinomas and sarcomas. HGF/SF, like many other angiogenic growth factors, employs heparan sulphate as co-receptor. The role of this interaction has not been completely defined but appears to be physiologically relevant. Thus the presence of heparin increases the potency of HGF/SF in experiments with cells in culture leading to elevated downstream signalling effects and, although not vital for the Met-HGF/SF interaction, heparin or heparan sulphate is essential for the activity of certain isoforms of HGF/SF, such as NK1 and NK2. Here, we summarize the progress made in understanding the interaction between heparin and heparan sulphate and NK1, NK2 and HGF/SF and we discuss their role in HGF/SF-Met signalling. PMID- 16709176 TI - Interaction of the guidance molecule Slit with cellular receptors. AB - Slits are large secreted glycoproteins characterized by an unusual tandem of four LRR (leucine-rich repeat) domains in their N-terminal half. Slit proteins were initially described as repulsive guidance cues in neural development, but it has become clear that they have additional important functions, for instance in the vasculature and immune system. Genetic studies have identified two types of cellular receptors for Slits: Robos (Roundabout) and the HS (heparan sulphate) proteoglycan syndecan. The intracellular signalling cascade downstream of Robo activation is slowly being elucidated, but the mechanism of transmembrane signalling by Robo has remained obscure. No active signalling role for syndecan has yet been demonstrated. Slit-HS interactions may be important for shaping the presumed Slit gradient or presenting Slit at its target cell surface. Recent studies have mapped the binding sites for Robos and HS/heparin to discrete Slit domains. Robos bind to the second LRR domain of Slit, whereas HS/heparin binds with very high affinity to the C-terminal portion of Slit. Slit activity is likely to be modulated by physiological proteolytic cleavage in the region separating the Robo and HS/heparin-binding sites. PMID- 16709177 TI - The biological relevance of chemokine-proteoglycan interactions. AB - Chemokines exert their biological activity through high-affinity interactions with cell-surface receptors, thereby activating specific signalling pathways, and a second low-affinity interaction with proteoglycans. Proteoglycans consist of a protein core, to which GAG (glycosaminoglycan) chains are attached. The GAGs are long, linear, sulphated and highly charged heterogeneous polysaccharides that are expressed throughout the body in different forms depending on the developmental or pathological state of the organ/organism. Mechanistically, the GAG interaction is thought to facilitate the retention of chemokines on cell surfaces, thereby forming a high local concentration required for cell activation. Recently, we demonstrated that certain chemokines require interactions with GAGs for their in vivo function. Additionally we have shown that chemokines oligomerize on immobilized GAGs, and this ability to form higher order oligomers has also been shown to be essential for the activity of certain chemokines in vivo. We believe that interference with the chemokine-GAG interaction provides a novel anti inflammatory strategy, exemplified by a variant of RANTES (regulated upon activation, normal T-cell expressed and secreted) that has abrogated GAG binding and oligomerization properties. PMID- 16709178 TI - Protein-GAG interactions: new surface-based techniques, spectroscopies and nanotechnology probes. AB - New approaches, rooted in the physical sciences, have been developed to gain a more fundamental understanding of protein-GAG (glycosaminoglycan) interactions. DPI (dual polarization interferometry) is an optical technique, which measures real-time changes in the mass of molecules bound at a surface and the geometry of the bound molecules. QCM-D (quartz crystal microbalance-dissipation), an acoustic technique, measures the mass and the viscoelastic properties of adsorbates. The FTIR (Fourier-transform IR) amide bands I, II and III, resulting from the peptide bond, provide insight into protein secondary structure. Synchrotron radiation CD goes to much shorter wavelengths than laboratory CD, allowing access to chromophores that provide insights into the conformation of the GAG chain and of beta-strand structures of proteins. To tackle the diversity of GAG structure, we are developing noble metal nanoparticle probes, which can be detected at the level of single particles and so enable single molecule biochemistry and analytical chemistry. These new approaches are enabling new insights into structure-function relationships in GAGs and together they will resolve many of the outstanding problems in this field. PMID- 16709179 TI - Computational approaches to the identification of heparin-binding sites on the surfaces of proteins. AB - The identification of heparin-binding sites is important for understanding the physiological function of many secreted proteins. Most of the experimental techniques for mapping these sites do not define them to atomic resolution. The use of automated docking methods can aid this process by facilitating both the design of experiments and visualization of their results. A method designed for a systematic search over the whole protein surface for heparin-binding sites, using heparin oligosaccharide structures as ligands, is described, with its validation and details of several published applications. The scope and limitations of this crude but effective computational chemistry method are discussed. PMID- 16709180 TI - Developing chemokine mutants with improved proteoglycan affinity and knocked-out GPCR activity as anti-inflammatory recombinant drugs. AB - The interaction of chemokines and GAGs (glycosaminoglycans) on endothelial surfaces is a crucial step for establishing a chemotactic gradient which leads to the functional presentation of chemokines to their GPCRs (G-protein-coupled receptors) and thus to activation of approaching leucocytes. Based on molecular modelling, biophysical investigations, cell-based and in vivo experiments, we have developed a novel concept for therapeutically interfering with chemokine-GAG interactions, namely dominant-negative chemokine mutants with improved GAG binding affinity and knocked-out GPCR activity. These recombinant proteins displace their wild-type chemokine counterparts from the natural proteoglycan co receptors without being able to activate leucocytes via GPCRs. Our mutant chemokines therefore represent the first protein-based GAG antagonists with high therapeutic potential in inflammatory diseases. PMID- 16709181 TI - Multiprotein signalling complexes: regional assembly on heparan sulphate. AB - Heparan sulphate (HS) is an abundant component of cell surfaces and the extracellular matrix. It binds to a wide variety of peptide growth factors, morphogens, chemokines and extracellular matrix proteins (e.g. fibronectin) and many of these interactions are essential for these effector proteins to transduce signals across the plasma membrane. The unique molecular design and flexibility of HS are essential for its ability to exert control over the cellular response to proteinaceous ligands. The clustering of sulphated sugar residues in a series of complex domains with variable sulphation patterns generates considerable diversity in the molecular fine structure of HS. This diversity reflects a high degree of selectivity in protein recognition and in the assembly of functional multiprotein complexes on the HS polymer chain. PMID- 16709182 TI - Insights into the role of heparan sulphate in fibroblast growth factor signalling. AB - Signalling from the FGFs (fibroblast growth factors) is crucial for the correct development and homoeostasis of a wide range of cells and tissues. The FGF/FGFR (FGF receptor) signalling system forms an important paradigm for HS (heparan sulphate)-binding proteins, as both the growth factor and receptor bind to HS, and HS or heparin is an absolute requirement for full signalling. The FGF signalling system has been extremely well structurally characterized, and details of each interaction involved in forming a ternary complex of FGF-FGFR-heparin have been elucidated. Recent work has focused on a more thorough understanding of the nature of the FGF-heparin complex in particular, demonstrating that FGFs preferentially bind to similar sites on the co-receptor, and that FGF-FGFR pairs show greater specificity for heparin sulphation patterns than individual FGFs. Further work has suggested that FGF-FGFR-heparin signalling complexes contain one molecule of heparin only, and that when longer fragments of heparin are used to form FGF-FGFR-heparin complexes, multiple complexes form upon the saccharide. These observations form the basis of a model where the range of interactions that FGFs and FGFRs can form with one another and with HS may lead to the formation of complexes with more than two FGFR units. Therefore HS will be crucial to FGF signalling from the initial signalling event to the formation of large receptor clusters. PMID- 16709183 TI - TSG-6: a pluripotent inflammatory mediator? AB - TSG-6 is a multifunctional protein that is up-regulated in many pathological and physiological contexts, where it plays important roles in inflammation and tissue remodelling. For example, it is a potent inhibitor of neutrophil migration and can modulate the protease network through inhibition of plasmin. TSG-6 binds a wide range of GAGs (glycosaminoglycans) [i.e. HA (hyaluronan), chondroitin 4 sulphate, dermatan sulphate, heparin and heparan sulphate] as well as a variety of protein ligands, where these interactions can influence the activities of TSG 6. For example, through its association with HA, TSG-6 can mediate HA cross linking via several different mechanisms, some of which promote leucocyte adhesion. Binding to heparin, however, enhances the ability of TSG-6 to potentiate the anti-plasmin activity of inter-alpha-inhibitor, which binds non covalently to TSG-6 via its bikunin chain. Furthermore, although HA and heparin interact with distinct sites on the Link module, the binding of heparin can inhibit subsequent interaction with HA. In addition, the interactions of TSG-6 with HA, heparin and at least some of its protein ligands are sensitive to pH. Therefore it seems that in different tissue micro-environments (characterized, for example, by pH and GAG content), TSG-6 could be partitioned into functional pools with distinct activities. PMID- 16709184 TI - The role of heparan sulphate proteoglycans in angiogenesis. AB - The presence of HS (heparan sulphate) proteoglycans on the cell surface and in the extracellular environment is critical to many physiological processes including the growth of new blood vessels from pre-existing vasculature (angiogenesis). A plethora of growth factors and their receptors, extracellular matrix molecules and enzymes bind to specific sites on the HS sugar chain. For example, HS proteoglycans have profound effects on the bioactivity of the key angiogenic factor VEGF (vascular endothelial growth factor) (VEGF(165)), affecting its diffusion, half-life and interaction with its tyrosine kinase receptors. A number of HS structural features that mediate the specific binding of VEGF(165), including sulphation requirements, have been determined. In parallel, zebrafish embryos were used as a vertebrate model system to study the role in vascular development of the biosynthetic enzymes that create these specific binding sites on HS. It was discovered that knockdown of one of the HS 6 O-sulphotransferases in zebrafish with morpholino antisense oligonucleotides reduced vascular branching and corresponded to changes in the HS structure. The roles of the extracellular 6-O-sulphatase enzymes, the sulfs, in vascular development are now being investigated. Both oligosaccharides and small molecule biosynthetic enzyme inhibitors could be valuable HS-based strategies for controlling aberrant angiogenesis in diseases as diverse as cancer and heart disease. PMID- 16709185 TI - Heparan sulphate requirement in platelet-derived growth factor B-mediated pericyte recruitment. AB - HS (heparan sulphate) plays a key role in angiogenesis, by interacting with growth factors required in the process. It has been proposed that HS controls the diffusion, and thus the availability, of platelet-derived growth factor B that is needed for pericyte recruitment around newly formed capillaries. The present paper summarizes our studies on the importance of HS structure in this regulatory process. PMID- 16709186 TI - Basic fibroblast growth factor: an extracellular mechanotransducer in articular cartilage? AB - Mechanical stimuli are important signals in articular cartilage, but what mediates them is unknown. We have shown that extracellular-signal-regulated kinase was activated on cutting and loading articular cartilage, and deduced that this was due to the release of bFGF (basic fibroblast growth factor) from the tissue. bFGF was shown to be extracellular, and by immunohistochemistry, was present in the pericellular matrix of articular chondrocytes attached to the heparan sulphate proteoglycan perlecan. We propose a novel mechanotransduction model, whereby pericellular bFGF, a short distance from the cell surface, becomes available to the cell surface tyrosine kinase receptors when articular cartilage is loaded. PMID- 16709187 TI - Heparin/heparan sulphate binding in the TGF-beta cytokine superfamily. AB - The TGF-beta (transforming growth factor-beta) cytokine superfamily in mammals contains some 30 members. These dimeric proteins are characterized by a strongly conserved cystine knot-based structure. They regulate the proliferation, differentiation and migration of many cell types, and therefore have important roles in morphogenesis, organogenesis, tissue maintenance and wound healing. Thus far, around one-quarter of these cytokines have been shown to bind to heparin and heparan sulphate. Well-established examples are the TGF-beta isoforms 1 and 2, and the BMPs (bone morphogenetic proteins) -2 and -4. In studies in my laboratory, we have shown that GDNF (glial-cell-line-derived neurotrophic factor) and its close relatives neurturin and artemin bind to heparin and heparan sulphate with high affinity. We have reported previously that binding of GDNF is highly dependent on the presence of 2-O-sulphate groups. More recently, we and others have been investigating the heparin/heparan sulphate-binding properties of BMP-7, which is a representative of a distinct BMP subgroup from that of BMPs -2 and -4. Interestingly, several of the various specific BMP antagonist proteins also bind to heparin and heparan sulphate. Much remains to be learnt about the nature and role of glycosaminoglycan interactions in the TGF-beta superfamily, but current work suggests that these cytokines do not share a single highly conserved heparin/heparan sulphate-binding site. PMID- 16709188 TI - Interferon and heparan sulphate. AB - In 1954, substances that protected cells from viral infection were discovered and named IFN (interferon). This family of cytokines, which were the first to be used in clinical therapy, is classified into type I and II IFNs. Type I mainly consists of IFNalpha and IFNbeta subtypes, which are structurally related and bind to a common receptor. IFNgamma, the sole type II IFN, is structurally unrelated, binds to a different receptor and, as a dimer, strongly interacts with HS (heparan sulphate). In addition to its antiviral activity, it modulates nearly all phases of immune and inflammatory responses. IFNgamma binding to HS controls the blood clearance, the subsequent tissue targeting and the local accumulation of the cytokine. It also regulates IFNgamma activity by a unique mechanism involving a controlled processing of the C-terminal peptide. The binding site encompasses an N-acetylated glucosamine-rich domain separating two highly sulphated sequences that each binds to one IFNgamma monomer. Based on this template, a set of glycoconjugate mimetics that would mimic the IFNgamma binding site has been synthesized. One of these molecules displays high affinity for the cytokine and inhibits binding to both HS and IFNgammaR (IFNgamma receptor), the cell-surface receptor. These results validate the HS structural determinants for IFNgamma recognition, and provide a new strategy to inhibit IFNgamma in a number of diseases in which the cytokine has been identified as a target. PMID- 16709189 TI - Gene duplication, exon gain and neofunctionalization of OEP16-related genes in land plants. AB - OEP16, a channel protein of the outer membrane of chloroplasts, has been implicated in amino acid transport and in the substrate-dependent import of protochlorophyllide oxidoreductase A. Two major clades of OEP16-related sequences were identified in land plants (OEP16-L and OEP16-S), which arose by a gene duplication event predating the divergence of seed plants and bryophytes. Remarkably, in angiosperms, OEP16-S genes evolved by gaining an additional exon that extends an interhelical loop domain in the pore-forming region of the protein. We analysed the sequence, structure and expression of the corresponding Arabidopsis genes (atOEP16-S and atOEP16-L) and demonstrated that following duplication, both genes diverged in terms of expression patterns and coding sequence. AtOEP16-S, which contains multiple G-box ABA-responsive elements (ABREs) in the promoter region, is regulated by ABI3 and ABI5 and is strongly expressed during the maturation phase in seeds and pollen grains, both desiccation-tolerant tissues. In contrast, atOEP-L, which lacks promoter ABREs, is expressed predominantly in leaves, is induced strongly by low-temperature stress and shows weak induction in response to osmotic stress, salicylic acid and exogenous ABA. Our results indicate that gene duplication, exon gain and regulatory sequence evolution each played a role in the divergence of OEP16 homologues in plants. PMID- 16709190 TI - The Arabidopsis SPA1 gene is required for circadian clock function and photoperiodic flowering. AB - Arabidopsis phytochrome A (phyA) regulates not only seed germination and seedling de-etiolation but also circadian rhythms and flowering time in adult plants. The SUPPRESSOR OF PHYA-105 (SPA1) acts as a negative regulator of phyA-mediated de etiolation of young seedlings, but its roles in adult plants have not yet been described. Here, we show that SPA1 is involved in regulating circadian rhythms and flowering time in plants. Under constant light, the abundance of SPA1 protein exhibited circadian regulation, whereas under constant darkness, SPA1 protein levels remained unchanged. These results indicate that the SPA1 protein is controlled by the circadian clock and light signals. In addition, the spa1-3 mutation slightly shortened the circadian period of CCA1, TOC1/PRR1 and SPA1 transcript accumulation under constant light. Phenotypic analysis showed that the spa1-3 mutant flowers early under short-day (SD) but not long-day (LD) conditions. Consistent with this finding, transcripts encoding flowering locus T (FT), which promotes flowering, increased in spa1-3 under only SD conditions, although the CONSTANS (CO) transcript level was not affected under either SD nor LD conditions. Our results indicate that SPA1 not only negatively controls phyA mediated signaling in seedlings, but also regulates circadian rhythms and flowering time in plants. PMID- 16709191 TI - Maize DBF1-interactor protein 1 containing an R3H domain is a potential regulator of DBF1 activity in stress responses. AB - The maize dehydration-responsive element (DRE)-binding factor, DBF1, is a member of the Apetala 2/Ethylene Response Factor transcription factors family and is involved in the regulation of the ABA-responsive gene rab17 through the DRE in an ABA-dependent pathway. In this study we analysed the functionality of DBF1 in abiotic stress responses and found that Arabidopsis plants over-expressing DBF1 were more tolerant to osmotic stress than control plants. In yeast two-hybrid analyses, DBF1 interacted with DBF1-interactor protein 1 (DIP1), a protein containing a conserved R3H single-strand DNA-binding domain. Subcellular localization of DIP1 showed that the protein fusion DIP1-Red Flourescent Protein (RFP) was mainly localized in the cytoplasm. However, after co-transformation of DBF1-GFP and DIP1-RFP, both proteins co-localized in the nucleus. Interestingly, when the N-terminal DBF1-GFP was co-expressed with DIP1-RFP, both proteins co localized predominantly in the cytoplasmic speckles observed for N-terminal DBF1 GFP fusion protein. These results clearly show in vivo interaction of DBF1 with DIP1 in the cell and that this interaction is necessary for the nuclear localization of DIP1 protein. Analysis of the regulatory effect of the DBF1 and DIP1 interaction on the maize rab17 promoter activity indicated that co transfection of DBF1 with DIP1 enhances promoter activity in the absence of ABA treatment. We suggest that the regulated association of DBF1 and DIP1 may control the levels of target gene expression during stress conditions. PMID- 16709192 TI - Altering glucosinolate profiles modulates disease resistance in plants. AB - Plant diseases are major contributing factors for crop loss in agriculture. Here, we show that Arabidopsis plants with high levels of novel glucosinolates (GSs) as a result of the introduction of single CYP79 genes exhibit altered disease resistance. Arabidopsis expressing CYP79D2 from cassava accumulated aliphatic isopropyl and methylpropyl GS, and showed enhanced resistance against the bacterial soft-rot pathogen Erwinia carotovora, whereas Arabidopsis expressing the sorghum CYP79A1 or over-expressing the endogenous CYP79A2 accumulated p hydroxybenzyl or benzyl GS, respectively, and showed increased resistance towards the bacterial pathogen Pseudomonas syringae. In addition to the direct toxic effects of GS breakdown products, increased accumulation of aromatic GSs was shown to stimulate salicylic acid-mediated defenses while suppressing jasmonate dependent defenses, as manifested in enhanced susceptibility to the fungus Alternaria brassicicola. Arabidopsis with modified GS profiles provide important tools for evaluating the biological effects of individual GSs and thereby show potential as biotechnological tools for the generation of plants with tailor-made disease resistance. PMID- 16709193 TI - TT8 controls its own expression in a feedback regulation involving TTG1 and homologous MYB and bHLH factors, allowing a strong and cell-specific accumulation of flavonoids in Arabidopsis thaliana. AB - The control of TT8 expression was investigated in this study, and it was demonstrated that it constitutes a major regulatory step in the specific activation of the expression of flavonoid structural genes. First, the GUS activity generated in planta from a TT8::uidA construct revealed cell-specific activation of the TT8 promoter consistent with the known involvement of the TT8 bHLH factor in proanthocyanidin, anthocyanin and mucilage biosynthesis. Moreover, the activity of this reporter construct was strongly affected in ttg1, TT2 overexpressers (OE), and PAP1-OE, suggesting interplay between TT2, PAP1, TTG1 and the activation of the TT8 promoter in planta. To further investigate the mechanisms involved, we used 35S::TT2-GR and 35S::TTG1-GR transgenic plants (expressing fusion proteins with the glucocorticoid receptor), as well as one hybrid experiments, to determine the direct effect of these factors on TT8 expression. Interestingly, in vivo binding of TT2 and PAP1 to the TT8 promoter was dependent on the simultaneous expression of TT8 or the homologous bHLH factors GL3 and EGL3. Consistent with these results, the activity of the TT8::uidA reporter was strongly affected in the seed endothelium of a tt8 mutant. Similarly, a strong decrease in the level of TT8 mRNA was detected in the siliques of a gl3 x egl3 mutant and in plants that express a dominant negative form of the PAP1 protein, suggesting that TT8 expression is controlled by different combinations of MYB and bHLH factors in planta. The importance of this positive feedback mechanism in the strong and specific induction of proanthocyanidin biosynthesis in the seed coat of Arabidopsis thaliana is discussed. PMID- 16709194 TI - AhSSK1, a novel SKP1-like protein that interacts with the S-locus F-box protein SLF. AB - The S-locus F-box (SLF/SFB) protein, recently identified as the pollen determinant of S-RNase-based self-incompatibility (SI) in Solanaceae, Scrophulariaceae and Rosaceae, has been proposed to serve as the subunit of an SCF (SKP1-CUL1-F-box) ubiquitin ligase and to target its pistil counterpart S RNase during the SI response. However, the underlying mechanism is still in dispute, and the putative SLF-binding SKP1-equivalent protein remains unknown. Here, we report the identification of AhSSK1, Antirrhinum hispanicumSLF interacting SKP1-like1, using a yeast two-hybrid screen against a pollen cDNA library. GST pull-down assays confirmed the SSK1-SLF interaction, and showed that AhSSK1 could connect AhSLF to a CUL1-like protein. AhSSK1, despite having a similar secondary structure to other SKP1-like proteins, appeared quite distinctive in sequence and unique in a phylogenetic analysis, in which no SSK1 ortholog could be predicted in the sequenced genomes of Arabidopsis and rice. Thus, our results suggest that the pollen-specific SSK1 could be recruited exclusively as the adaptor of putative SCF(SLF) in those plants with S-RNase based SI, providing an important clue to dissecting the function of the pollen determinant. PMID- 16709195 TI - A B-lectin receptor kinase gene conferring rice blast resistance. AB - Rice blast, caused by the fungal pathogen Magnaporthe grisea, is one of the most devastating diseases in rice worldwide. The dominant resistance gene, Pi-d2 [previously named Pi-d(t)2], present in the rice variety Digu, confers gene-for gene resistance to the Chinese blast strain, ZB15. Pi-d2 was previously mapped close to the centromere of chromosome 6. In this study, the Pi-d2 gene was isolated by a map-based cloning strategy. Pi-d2 encodes a receptor-like kinase protein with a predicted extracellular domain of a bulb-type mannose specific binding lectin (B-lectin) and an intracellular serine-threonine kinase domain. Pi d2 is a single-copy gene that is constitutively expressed in the rice variety Digu. Transgenic plants carrying the Pi-d2 transgene confer race-specific resistance to the M. grisea strain, ZB15. The Pi-d2 protein is plasma membrane localized. A single amino acid difference at position 441 of Pi-d2 distinguishes resistant and susceptible alleles of rice blast resistance gene Pi-d2. Because of its novel extracellular domain, Pi-d2 represents a new class of plant resistance genes. PMID- 16709196 TI - Gene expression profiles of Arabidopsis Cvi seeds during dormancy cycling indicate a common underlying dormancy control mechanism. AB - Physiologically dormant seeds, like those of Arabidopsis, will cycle through dormant states as seasons change until the environment is favourable for seedling establishment. This phenomenon is widespread in the plant kingdom, but has not been studied at the molecular level. Full-genome microarrays were used for a global transcript analysis of Arabidopsis thaliana (accession Cvi) seeds in a range of dormant and dry after-ripened states during cycling. Principal component analysis of the expression patterns observed showed that they differed in newly imbibed primary dormant seeds, as commonly used in experimental studies, compared with those in the maintained primary and secondary dormant states that exist during cycling. Dormant and after-ripened seeds appear to have equally active although distinct gene expression programmes, dormant seeds having greatly reduced gene expression associated with protein synthesis, potentially controlling the completion of germination. A core set of 442 genes were identified that had higher expression in all dormant states compared with after ripened states. Abscisic acid (ABA) responsive elements were significantly over represented in this set of genes the expression of which was enhanced when multiple copies of the elements were present. ABA regulation of dormancy was further supported by expression patterns of key genes in ABA synthesis/catabolism, and dormancy loss in the presence of fluridone. The data support an ABA-gibberelic acid hormone balance mechanism controlling cycling through dormant states that depends on synthetic and catabolic pathways of both hormones. Many of the most highly expressed genes in dormant states were stress related even in the absence of abiotic stress, indicating that ABA, stress and dormancy responses overlap significantly at the transcriptome level. PMID- 16709197 TI - RFI2, a RING-domain zinc finger protein, negatively regulates CONSTANS expression and photoperiodic flowering. AB - The red and far-red light-absorbing phytochromes interact with the circadian clock, a central oscillator that sustains a 24-h period, to measure accurately seasonal changes in day-length and regulate the expression of several key flowering genes. The interactions and subsequent signalling steps upstream of the flowering genes such as CONSTANS (CO) and FLOWERING LOCUS T (FT) remain largely unknown. We report here that a photomorphogenic mutant, red and far-red insensitive 2-1 ( rfi2-1), flowered early particularly under long days. The rfi2 1 mutation also enhanced the expression of CO and FT under day/night cycles or constant light. Both co-2 and gigantea-2 (gi-2) were epistatic to rfi2-1 in their flowering responses. The gi-2 mutation was also epistatic to the rfi2-1 mutation in the expression of CO and hypocotyl elongation. However, the rfi2-1 mutation did not affect the expression of GI, a gene that mediates between the circadian clock and the expression of CO. Like many other flowering genes, the expression of RFI2 oscillated under day/night cycles and was rhythmic under constant light. The amplitude of the rhythmic expression of RFI2 was significantly reduced in phyB-9 or lhy-20 plants, and was also affected by the gi-2 mutation. As previously reported, the gi-2 mutation affects the period length and amplitude of CCA1 and LHY expression, and GI may act through a feedback loop to maintain a proper circadian function. We propose a regulatory step in which RFI2 represses the expression of CO, whereas GI may maintain the proper expression of RFI2 through its positive action on the circadian clock. The regulatory step serves to tune the circadian outputs that control the expression of CO and photoperiodic flowering. PMID- 16709199 TI - The proteasome-dependent degradation of CKB4 is regulated by the Arabidopsis biological clock. AB - Most organisms have evolved an internal timing mechanism, the circadian clock, that is able to generate and maintain 24 h rhythmic oscillation in molecular, biochemical and metabolic activities. In Arabidopsis, the clock-dependent synchronization of physiology with the environment is essential for successful growth and development. The mechanisms of the Arabidopsis clockwork have been described as transcriptional feedback loops at the core of the oscillator. However, an increasing body of evidence points towards a key role of post translational regulation of clock components as an essential mechanism of circadian function. Here, we identify CKB4, a CK2 regulatory subunit, as a component of the Arabidopsis circadian system. We demonstrate that the nuclear localized CKB4 protein exists in vivo as different isoforms, resulting from phosphorylation on serine residues. Our findings show that the phosphorylated isoforms are the preferred substrate for ubiquitination and degradation by the proteasome pathway. We provide evidence of the involvement of the biological clock in the circadian regulation of CKB4 protein abundance, which itself is important for an accurate control of circadian period by the clock. Overexpression of CKB4 results in elevated CK2 overall activity and period shortening of clock-controlled genes peaking at different phase angles. Restriction of CKB4 protein phosphorylation and/or degradation to specific phases within the circadian cycle might provide the cell with a fine-tuning mechanism to selectively regulate the CK2 phosphorylation activity on specific substrates. PMID- 16709198 TI - Over-expression of SOB5 suggests the involvement of a novel plant protein in cytokinin-mediated development. AB - Cytokinins are a class of phytohormones that play a critical role in plant growth and development. sob5-D, an activation-tagging mutant, shows phenotypes typical of transgenic plants expressing the Agrobacterium tumefaciens isopentenyltransferase (ipt) gene that encodes the enzyme catalyzing the first step of cytokinin biosynthesis. The sob5-D mutant phenotypes are caused by over expression of a novel gene, SOB5. Sequence analysis places SOB5 in a previously uncharacterized family of plant-specific proteins. A translational fusion between SOB5 and the green fluorescent protein reporter was localized in the cytoplasm as well as associated with the plasma membrane when transiently expressed in onion epidermal cells. Analysis of transgenic plants harboring an SOB5:SOB5-beta glucuronidase (GUS) translational fusion under the control of the SOB5 promoter region showed GUS activity in vegetative tissues (hydathodes and trichomes of leaves, shoot meristems and roots) as well as in floral tissues (pistil tips, developing anthers and sepal vasculature). Cytokinin quantification analysis revealed that adult sob5-D plants accumulated higher levels of trans-zeatin riboside, trans-zeatin riboside monophosphate and isopentenyladenine 9-glucoside when compared to the wild-type. Consistent with this result, AtIPT3 and AtIPT7 were found to be up-regulated in a tissue-specific manner in sob5-D mutants. Physiological analysis of the sob5-D mutant demonstrated reduced responsiveness to exogenous cytokinin in both root-elongation and callus-formation assays. Taken together, our data suggest a role for the novel gene SOB5 in cytokinin-mediated plant development. PMID- 16709200 TI - The Arabidopsis metal tolerance protein AtMTP3 maintains metal homeostasis by mediating Zn exclusion from the shoot under Fe deficiency and Zn oversupply. AB - Zinc ions are required to maintain the biological activity of numerous proteins. However, when mislocalized or accumulated in excess, Zn(2+) ions are toxic because of adventitious binding to proteins and displacement of other metal ions, among them Fe(2+), from their binding sites. Heterologous expression of a previously uncharacterized Arabidopsis thaliana metal tolerance protein, MTP3, in the zrc1 cot1 mutant of budding yeast restores tolerance to, and cellular accumulation of, zinc and cobalt. An MTP3-GFP fusion protein localizes to the vacuolar membrane when expressed in Arabidopsis. Ectopic over-expression of MTP3 increases Zn accumulation in both roots and rosette leaves of A. thaliana, and enhances Zn tolerance. Exposure of wild-type plants to high but non-toxic concentrations of Zn or Co, or Fe deficiency, strongly induce MTP3 expression specifically in epidermal and cortex cells of the root hair zone. Silencing of MTP3 by RNA interference causes Zn hypersensitivity and enhances Zn accumulation in above-ground organs of soil-grown plants and of seedlings exposed to excess Zn or to Fe deficiency. Our data indicate that, in wild-type A. thaliana, the AtMTP3 protein contributes to basic cellular Zn tolerance and controls Zn partitioning, particularly under conditions of high rates of Zn influx into the root symplasm. PMID- 16709201 TI - Identification and characterization of Arabidopsis gibberellin receptors. AB - Three gibberellin (GA) receptor genes (AtGID1a, AtGID1b and AtGID1c), each an ortholog of the rice GA receptor gene (OsGID1), were cloned from Arabidopsis, and the characteristics of their recombinant proteins were examined. The GA-binding activities of the three recombinant proteins were confirmed by an in vitro assay. Biochemical analyses revealed similar ligand selectivity among the recombinants, and all recombinants showed higher affinity to GA(4) than to other GAs. AtGID1b was unique in its binding affinity to GA(4) and in its pH dependence when compared with the other two, by only showing binding in a narrow pH range (pH 6.4 7.5) with 10-fold higher affinity (apparent K(d) for GA(4) = 3 x 10(-8) m) than AtGID1a and AtGID1c. A two-hybrid yeast system only showed in vivo interaction in the presence of GA(4) between each AtGID1 and the Arabidopsis DELLA proteins (AtDELLAs), negative regulators of GA signaling. For this interaction with AtDELLAs, AtGID1b required only one-tenth of the amount of GA(4) that was necessary for interaction between the other AtGID1s and AtDELLAs, reflecting its lower K(d) value. AtDELLA boosted the GA-binding activity of AtGID1 in vitro, which suggests the formation of a complex between AtDELLA and AtGID1-GA that binds AtGID1 to GA more tightly. The expression of each AtGID1 clone in the rice gid1-1 mutant rescued the GA-insensitive dwarf phenotype. These results demonstrate that all three AtGID1s functioned as GA receptors in Arabidopsis. PMID- 16709203 TI - Isolation of precise plastid deletion mutants by homology-based excision: a resource for site-directed mutagenesis, multi-gene changes and high-throughput plastid transformation. AB - We describe a simple and efficient homology-based excision method to delete plastid genes. The procedure allows one or more adjacent plastid genes to be deleted without the retention of a marker gene. We used aadA-based transformation to duplicate a 649 bp region of plastid DNA corresponding to the atpB promoter region. Efficient recombination between atpB repeats deletes the intervening foreign genes and 1,984 bp of plastid DNA (co-ordinates 57,424-59,317) containing the rbcL gene. Only five foreign bases are present in DeltarbcL plants illustrating the precision of homology-based excision. Sequence analysis of non functional rbcL-related sequences in DeltarbcL plants indicated an extra plastidic origin. Mutant DeltarbcL plants were heterotrophic, pale-green and contained round plastids with reduced amounts of thylakoids. Restoration of autotrophy and leaf pigmentation following aadA-based transformation with the wild-type rbcL gene ruled out mutations in other genes. Excision and re-use of aadA shows that, despite the multiplicity of plastid genomes, homology-based excision ensures complete removal of functional aadA genes. Rescue of the DeltarbcL mutation and autotrophic growth stabilizes transgenic plastids in heteroplasmic transformants following antibiotic withdrawal, enhancing the overall efficiency of plastid transformation. Unlike the available set of homoplasmic knockout mutants in 25 plastid genes, the rbcL deletion mutant isolated here is readily transformed with the efficient aadA marker gene. This improvement in deletion design facilitates advanced studies that require the isolation of double mutants in distant plastid genes and the replacement of the deleted locus with site-directed mutant alleles and is not easily achieved using other methods. PMID- 16709202 TI - Two-hybrid protein-protein interaction analysis in Arabidopsis protoplasts: establishment of a heterodimerization map of group C and group S bZIP transcription factors. AB - In vivo protein-protein interactions are frequently studied by means of yeast two hybrid analysis. However, interactions detected in yeast might differ considerably in the plant system. Based on GAL4 DNA-binding (BD) and activation domains (AD) we established an Arabidopsis protoplast two-hybrid (P2H) system. The use of Gateway-compatible vectors enables the high-throughput screening of protein-protein interactions in plant cells. The efficiency of the system was tested by examining the homo- and heterodimerization properties of basic leucine zipper (bZIP) transcription factors. A comprehensive heterodimerization matrix of Arabidopsis thaliana group C and group S bZIP transcription factors was generated by comparing the results of yeast and protoplast two-hybrid experiments. Surprisingly, almost no homodimerization but rather specific and selective heterodimerization was detected. Heterodimers were preferentially formed between group C members (AtbZIP9, -10, -25, -63) and members of group S1 (AtbZIP1, -2, 11, -44, -53). In addition, significant but low-affinity interactions were detected inside group S1, S2 or C AtbZIPs, respectively. As a quantitative approach, P2H identified weak heterodimerization events which were not detected in the yeast system. Thus, in addition to cell biological techniques, P2H is a valuable tool for studying protein-protein interaction in living plant cells. PMID- 16709205 TI - Late miscarriage and preterm birth after treatment with clindamycin: a randomised consent design study according to Zelen. AB - OBJECTIVE: To screen for bacterial vaginosis (BV) and to investigate the effect of treatment with vaginal clindamycin in order to observe the effect on late miscarriage and delivery prior to 37 completed weeks (primary outcome). DESIGN: Randomised consent design for clinical trials according to Zelen. SETTING: Southeast region of Sweden. POPULATION: A total of 9025 women were screened in early pregnancy. METHODS: A total of 819 women with a Nugent score of 6 and above were considered to have BV and treated according to Zelen allocation. The incidence of late miscarriage and spontaneous (noniatrogenic) preterm birth was assessed. MAIN OUTCOME MEASURES: Late miscarriage and spontaneous preterm delivery before 37 weeks. RESULTS: Therapy with vaginal clindamycin had no significant impact on the incidence of spontaneous preterm delivery prior to 37 completed weeks; OR 0.90, 95% CI 0.40-2.02 (primary outcome variable). However, only 1 of 11 women in the treatment group versus 5 of 12 in the control group delivered prior to 33 completed weeks; OR 0.14, 95% CI 0.02-0.95. Treatment was associated with 32 days longer gestation for the 23 participants who had late miscarriage or spontaneous preterm birth (P= 0.024, Mann-Whitney U test) and significantly fewer infants had a birthweight below 2,500 g (secondary outcome). A follow up of infants born preterm 4 years postnatally indicated that extending gestational age did not increase the number of sequelae. CONCLUSIONS: Clindamycin vaginal cream therapy was associated with significantly prolonged gestation and reduced cost of neonatal care in women with BV. Early screening for BV and treatment with clindamycin saved approximately 27 euro per woman. PMID- 16709204 TI - Regimens of misoprostol with mifepristone for early medical abortion: a randomised trial. AB - OBJECTIVE: To compare the efficacy, adverse effects and acceptability of the three most common misoprostol regimens used with mifepristone for medical abortion. DESIGN: Randomised nonblinded trial. SETTING: Three clinics associated with major research universities in Canada; two in major urban areas and one in a periurban area. POPULATION: Women of reproductive age. METHODS: Consenting women presenting for abortion services with gestations less than 56 days and who met inclusion criteria were given 200 mg mifepristone orally and then randomised into three misoprostol study groups: (group I) 400 micrograms of oral misoprostol, (group II) 600 micrograms of oral misoprostol, and (group III) 800 micrograms of vaginal misoprostol. Misoprostol was self-administered at home 24-48 hours following mifepristone, and participants were instructed to take a second similar misoprostol dose at 24 hours after the initial dose if bleeding was less than a normal menstrual period. MAIN OUTCOME MEASURES: Successful abortion without surgery was 94.1%, with no significant differences across the three study groups (94.7% in group I, 93.4% in group II, and 94.3% in group III; P= 0.975). RESULTS: Efficacy and adverse effects did not differ significantly across the three study groups. Pain increased significantly across the study and the gestational age groups and was associated with lower acceptability. CONCLUSIONS: There appears to be a range of safe and effective options for early medical abortion with mifepristone including a choice between oral and vaginal administration of misoprostol. PMID- 16709206 TI - Antenatal fear of childbirth and its association with subsequent caesarean section and experience of childbirth. AB - OBJECTIVE: To investigate the prevalence of fear of childbirth in a nationwide sample and its association with subsequent rates of caesarean section and overall experience of childbirth. DESIGN: A prospective study using between-group comparisons. SETTING: About 600 antenatal clinics in Sweden. SAMPLE: A total of 2,662 women recruited at their first visit to an antenatal clinic during three predetermined weeks spread over 1 year. METHODS: Postal questionnaires at 16 weeks of gestation (mean) and 2 months postpartum. Women with fear of childbirth, defined as 'very negative' feelings when thinking about the delivery in second trimester and/or having undergone counselling because of fear of childbirth later in pregnancy, were compared with those in the reference group without these characteristics. MAIN OUTCOME MEASURES: Elective and emergency caesarean section and overall childbirth experience. RESULTS: In total 97 women (3.6%) had very negative feelings and about half of them subsequently underwent counselling. In addition, 193 women (7.2%) who initially had more positive feelings underwent counselling later in pregnancy. In women who underwent counselling, fear of childbirth was associated with a three to six times higher rate of elective caesarean sections but not with higher rates of emergency caesarean section or negative childbirth experience. Very negative feelings without counselling were not associated with an increased caesarean section rate but were associated with a negative birth experience. CONCLUSIONS: At least 10% of pregnant women in Sweden suffer from fear of childbirth. Fear of childbirth in combination with counselling may increase the rate of elective caesarean sections, whereas fear without treatment may have a negative impact on the subsequent experience of childbirth. PMID- 16709208 TI - Maternal haemodynamics in pre-eclampsia compared with normal pregnancy during caesarean delivery. AB - OBJECTIVE: To determine how pre-eclampsia modifies maternal haemodynamics during caesarean delivery. DESIGN: Prospective study. SETTING: Tampere University Hospital, Finland. POPULATION: Ten pre-eclamptic parturients and ten healthy parturients with uncomplicated pregnancies scheduled for elective caesarean section under spinal anaesthesia. METHODS: Haemodynamic parameters were assessed by whole-body impedance cardiography noninvasively. MAIN OUTCOME MEASURES: Stroke index (SI), heart rate (HR), cardiac index (CI), systemic vascular resistance index (SVRI) and mean arterial pressure (MAP) were recorded before operation, continuously during caesarean section, during the period of dissipation of anaesthesia and on the second to fifth postpartum day. RESULTS: Baseline haemodynamics in women with pre-eclampsia differed significantly from healthy women in higher SVRI and MAP and lower SI and CI. In women with pre-eclampsia, preload infusion increased both SI and HR, causing a significant rise in CI, while in healthy parturients, only HR rose. In both the groups, spinal blockade reduced SVRI but CI remained stable. At the moment of delivery, CI increased in both groups. In uncomplicated pregnancies, both SI and HR increased, but in women with pre-eclampsia, SI was not altered and the rise in CI was due to an increase in HR only. After the reversal of anaesthesia, haemodynamics in the control group returned to baseline values, whereas in women with pre-eclampsia, SI and CI fell to levels that were significantly lower than the levels observed before surgery. CONCLUSIONS: In women with pre-eclampsia, inability to increase SI at the moment of delivery may suggest dysfunction of the left ventricle to adapt to volume load caused by delivery and prompts concern for the increased risk of pulmonary oedema. PMID- 16709207 TI - Caesarean section on request: a comparison of obstetricians' attitudes in eight European countries. AB - OBJECTIVE: To explore the attitudes of obstetricians to perform a caesarean section on maternal request in the absence of medical indication. DESIGN: Cluster sampling cross-sectional survey. SETTING: Neonatal Intensive Care Unit (NICU) associated maternity units in eight European countries. POPULATION: Obstetricians with at least 6 months clinical experience. METHODS: NICU-associated maternity units were chosen by census in Luxembourg, Netherlands and Sweden and by geographically stratified random sampling in France, Germany, Italy, Spain and UK. An anonymous, self-administered questionnaire was used for data collection. MAIN OUTCOME MEASURES: Obstetricians' willingness to perform a caesarean section on maternal request. RESULTS: One hundred and five units and 1,530 obstetricians participated in the study (response rates of 70 and 77%, respectively). Compliance with a hypothetical woman's request for elective caesarean section simply because it was 'her choice' was lowest in Spain (15%), France (19%) and Netherlands (22%); highest in Germany (75%) and UK (79%) and intermediate in the remaining countries. Using weighted multivariate logistic regression, country of practice (P<0.001), fear of litigation (P= 0.004) and working in a university affiliated hospital (P= 0.001) were associated with physicians' likelihood to agree to patient's request. The subset of female doctors with children was less likely to agree (OR 0.29, 95% CI 0.20-0.42). CONCLUSIONS: The differences in obstetricians' attitudes are not founded on concrete medical evidence. Cultural factors, legal liability and variables linked to the specific perinatal care organisation of the various countries play a role. Greater emphasis should be placed on understanding the motivation, values and fears underlying a woman's request for elective caesarean delivery. PMID- 16709209 TI - Detection of malformations in chromosomally normal fetuses by routine ultrasound at 12 or 18 weeks of gestation-a randomised controlled trial in 39,572 pregnancies. AB - OBJECTIVE: To compare the antenatal detection rate of malformations in chromosomally normal fetuses between a strategy of offering one routine ultrasound examination at 12 gestational weeks (gws) and a strategy of offering one routine examination at 18 gws. DESIGN: Randomised controlled trial. SETTING: Multicentre trial including eight hospitals. POPULATION: A total of 39,572 unselected pregnant women. METHODS: Women were randomised either to one routine ultrasound scan at 12 (12-14) gws including nuchal translucency (NT) measurement or to one routine scan at 18 (15-22) gws. Anomaly screening was performed in both groups following a check-list. A repeat scan was offered in the 12-week scan group if the fetal anatomy could not be adequately seen at 12-14 gws or if NT was >or=3.5 mm in a fetus with normal or unknown chromosomes. MAIN OUTCOME MEASURES: Antenatal detection rate of malformed fetuses. RESULTS: The antenatal detection rate of fetuses with a major malformation was 38% (66/176) in the 12-week scan group and 47% (72/152) in the 18-week scan group (P= 0.06). The corresponding figures for detection at <22 gws were 30% (53/176) and 40% (61/152) (P= 0.07). In the 12-week scan group, 69% of fetuses with a lethal anomaly were detected at a scan at 12-14 gws. CONCLUSIONS: None of the two strategies for prenatal diagnosis is clearly superior to the other. The 12-week strategy has the advantage that most lethal malformations will be detected at <15 gws, enabling earlier pregnancy termination. The 18-week strategy seems to be associated with a slightly higher detection rate of major malformations, although the difference was not statistically significant. PMID- 16709210 TI - Routine ultrasound examination at 12 or 18 gestational weeks for prenatal detection of major congenital heart malformations? A randomised controlled trial comprising 36,299 fetuses. AB - OBJECTIVE: To compare the rate of prenatal diagnosis of heart malformations between two policies of screening for heart malformations. DESIGN: Randomised controlled trial. SETTING: Six university hospitals, two district general hospitals. SAMPLE: A total of 39 572 unselected pregnancies randomised to either policy. METHODS: The 12-week policy implied one routine scan at 12 weeks including measurement of nuchal translucency (NT), and the 18-week policy implied one routine scan at 18 weeks. Fetal anatomy was scrutinised using the same check list in both groups, and in both groups, indications for fetal echocardiography were ultrasound findings of any fetal anomaly, including abnormal four-chamber view, or other risk factors for heart malformation. In the 12-week scan group, NT >or=3.5 mm was also an indication for fetal echocardiography. MAIN OUTCOME MEASURE: Prenatal diagnosis of major congenital heart malformation. RESULTS: In the 12-week scan group, 7 (11%) of 61 major heart malformations were prenatally diagnosed versus 9 (15%) of 60 in the 18-week scan group (P= 0.60). In four (6.6%) women in the 12-week scan group, the routine scan was the starting point for investigations resulting in a prenatal diagnosis versus in 9 (15%) women in the 18-week scan group (P=0.15). The diagnosis was made 2) moles, particularly in the absence of a positive family history, may be androgenetic rather than biparental. These data suggest that some women have a specific liability for having AnCHM. Making the distinction between a biparental or an androgenetic origin of recurrent moles is of relevance for counselling and when considering therapeutic options. Therefore, we propose that all recurrent moles should be investigated using molecular techniques. PMID- 16709218 TI - Uterine rupture and labour after a previous low transverse caesarean section. AB - This study reviewed all cases of complete uterine rupture (UR) in pregnancy during the decade 1993-2002. In 69,412 deliveries, 5,320 women had a single prior caesarean section. Of these, 4,021 had a trial of labour and 3,129 (77.8%) delivered vaginally. In nine (0.22%) cases, the previous transverse scar ruptured during labour. None of these nine ruptures resulted in maternal or fetal death, peripartum hysterectomy or fetal morbidity. In our practice, a trial of labour in women with a previous low transverse caesarean is associated with a high rate of vaginal delivery and a low rate of UR. PMID- 16709220 TI - Predicting the impact of in vitro fertilisation and other forms of assisted conception on perinatal and infant mortality in England and Wales: examining the role of multiplicity. AB - The increased risk of perinatal and infant mortality observed among in vitro fertilisation (IVF) births and other assisted conception births is thought to be largely attributable to multiplicity. Using mortality statistics and estimates of the proportion of births following infertility treatment, we predicted the excess stillbirths and infant deaths associated with twins and higher order births resulting from assisted conception in England and Wales. According to our results, approximately 73 deaths could have been avoided in 2001 if all IVF infants had been born as singletons or as naturally occurring monozygotic twins, equating to a population attributable risk fraction of around 1% for perinatal and infant deaths. If we include all types of assisted conception, this figure was estimated to be around 4% of deaths-more than 220 perinatal and infant deaths in 2001. We confirm the public health importance of multiple births associated with assisted conception. PMID- 16709219 TI - The fetal sex ratio and metabolic, biochemical, haematological and clinical indicators of severity of hyperemesis gravidarum. AB - The association between female fetal sex and hyperemesis gravidarum is well established in European and North American populations. The association between female fetuses and severity of hyperemesis remains uncertain. A retrospective study based on case notes review of 166 Asian women hospitalised for hyperemesis was performed. Female fetuses were significantly associated with hyperemesis in our population (P= 0.004, OR 1.6, 95% CI 1.2-2.2) as well as associated with severe ketonuria and high urea. When both severe ketonuria and high urea were present at initial hospital admission for hyperemesis, 83% (95% CI 66-93) of the fetuses were female. PMID- 16709221 TI - Priapism of the clitoris. PMID- 16709222 TI - Hook-wire insertion facilitates the excision of scar endometriosis. PMID- 16709223 TI - Follow up of a case of extreme birth asphyxia. PMID- 16709226 TI - Adrenomedullin regulates expressions of transforming growth factor-beta1 and beta1-induced matrix metalloproteinase-2 in hepatic stellate cells. AB - Adrenomedullin (AM), a peptide isolated from human pheochromocytoma, can be produced and secreted by various types of cells including hepatic stellate cells (HSCs), and its possible role in HSCs is not clear now. In the present study, the interactive regulation between transforming growth factor (TGF)-beta1 and AM and the effect of AM on TGF-beta1-induced matrix metalloproteinase (MMP)-2 expression in HSCs were investigated. TGF-beta1 and AM inhibited gene transcript level mutually (real-time reverse transcription-polymerase chain reaction). AM suppressed the protein expression level of TGF-beta1 (Western blot), but TGF beta1 might have no effect on AM secretion level. MMP-2 protein expression in HSCs was increased in response to TGF-beta1, and upregulation of MMP-2 expression stimulated with TGF-beta1 was suppressed by AM in dose-dependent manner (Western blot). AM decreased the phosphorylation level of extracellular signal-regulated kinase (ERK) in HSCs treated with TGF-beta1, and TGF-beta1-induced MMP-2 expression was suppressed by adding Mitogen-activated protein Kinase/ERK (MEK) inhibitor U(0126) (Western blot)(.) Our results suggest that AM may intervene the activation of HSCs by inhibiting TGF-beta1 production and TGF-beta1-induced MMP-2 expression; AM may suppress the upregulation of MMP-2 expression induced by TGF beta1 partially through ERK pathway. PMID- 16709225 TI - Tumour-associated macrophages and melanoma tumourigenesis: integrating the complexity. AB - When the body discovers a tumour cell (foreign antigen), several kinds of mechanisms and cells operate in what is called an immune response. The latter has evolved into two mechanisms: non-specific immunity and specific immunity, which are closely linked to and influence each other. The former represents the first line of defence against neoplastic cells. The adaptive (specific) immunity is orchestrated by antigen-specific T and B lymphocytes. The effector cells of innate immunity include granulocytes, macrophages and natural killer cells. Among these cells, macrophages represent the most important part of innate immunity against tumours. Tumour-associated macrophages (TAMs) are important antigen presenting cells and as such an understanding of their interactions with tumour cells gives insights into novel therapeutic strategies. In tumours, the effect of TAMs is the outcome of their two concomitantly competing interactions: tumour growth reduction and tumour growth promotion. The macrophage (TAMs) content of melanoma ranges from 0 to 30% and their density increases with increasing tumour thickness. The melanoma cells and TAMs seem to interact with each other through the release of soluble factors that either prevent or enhance tumour growth. For instance, syngeneic macrophages from tumour-bearing mice can inhibit melanoma growth in the nude mice more than the control macrophages. Alternatively, metastatic B16 melanoma cells can produce some macrophage cytotoxic substances that help tumour cells not only escape the host immunosurveillance system but also prevent distant metastasis. Together, these observations suggest opposing effects for these soluble factors in melanoma. To date, little is available in the literature about the interactions between TAMs and melanoma cells. This viewpoint not only tries to examine these interactions but also provides relevant speculations. PMID- 16709227 TI - Effects of transforming growth factor-beta in the development of inflammatory pseudotumour-like lesions in a murine model. AB - Alterations in transforming growth factor (TGF)-beta signalling have been frequently implicated in human cancer, and an important mechanism underlying its pro-oncogenic nature is suppression of the host antitumour immune response. Considering the immunosuppressive effect of TGF-beta, we asked whether human tumour cells, known to secrete TGF-beta in culture, would survive and grow when implanted into the peritoneal cavity of immunocompetent mice. Therefore, we developed a xenogeneic model where mice were intraperitoneally (i.p.) injected with a TGF-beta-secreting human colorectal adenocarcinoma cell line, LISP-A10. Although animals did not develop macroscopic tumours, the recovery and isolation of human tumour cells was achieved when an inflammatory environment was locally induced by the administration of complete Freund's adjuvant (CFA). This procedure significantly increased TGF-beta concentrations in the peritoneal fluid and was accompanied by impaired activation of the host-specific immune response against LISP-A10 cells. Furthermore, inflammatory lesions resembling human inflammatory pseudotumours (IPTs) were observed on the surface of i.p. organs. These lesions could be induced by either injection of LISP-A10 cells, cells-conditioned medium or recombinant TGF-beta but only after administration of CFA. In addition, host cyclooxygenase-2 and kinin receptors played an important role in the induction of TGF-beta-mediated IPT-like lesions in our experimental model. PMID- 16709228 TI - TGFbeta1 induces epithelial-mesenchymal transition, but not myofibroblast transdifferentiation of human kidney tubular epithelial cells in primary culture. AB - The origin and fate of renal interstitial myofibroblasts (MFs), the effector cells of renal fibrosis, are still debated. Experimental evidence suggests that renal MFs derive from tubular epithelial cells throughout the epithelial mesenchymal transition (EMT) process. Primary human tubular epithelial cells (HUTECs) were cultured for 4 and 6 days on plastic or type I collagen-coated plates with 1, 5, 10 and 50 ng/ml of transforming growth factor beta1 (TGFbeta1). The EMT process was monitored by morphology and immunophenotyping for alphaSMA, cytokeratin 8-18, E-cadherin, vimentin and collagen III. Quantitative comparative RT/PCR and real-time PCR were used to evaluate the expression of collagen III and IV, fibronectin, tenascin, MMP-2, CTGF, E-cadherin and cadherin 11 genes, as well as those of the Smad signalling pathway. TGFbeta1 was found capable of reactivating the mesenchymal programme switched off during tubulogenesis, but it induced no de novo expression of alphaSMA gene or myofibroblast phenotype. We demonstrate that the EMT process is conditioned by the extracellular matrix and characterized by TGFbeta1-driven Smad3 downregulation. Our study results suggest that TGFbeta1 could function as a classic embryonal inducer, initiating a cascade of de-differentiating events that might be further controlled by other factors in the cellular environment. PMID- 16709229 TI - Trypanosoma cruzi: clones isolated from the Colombian strain, reproduce the parental strain characteristics, with ubiquitous histotropism. AB - Clonal histotropism and biological characters of five clones isolated during the early acute phase of the infection of Swiss mice with the Colombian strain of Trypanosoma cruzi (T. cruzi I), Biodeme Type III, were investigated. Clones were isolated from mice at the 10th and the 30th day of infection with the Colombian strain. Isolation was performed by micromanipulation and injection of one trypomatigote blood form into newborn mice, followed by passages into suckling mice for obtaining the inocula for the experimental groups. Mice infected with parental strain were also studied. All the clones have shown the basic characteristics of Biodeme Type III, with the same patterns of parasitemia, tissue tropism, morphological characters and isoenzymic profiles, such as the parental strain. Histotropism was most intense to myocardium and skeletal muscles, with intense lesions found in the advanced phase (20th to 30th day of infection). Both parental strain and the clones were seen to parasitize several organs and tissues; amastigote nests were identified in the cytoplasm of macrophages, adipose cells, smooth muscle of intestinal wall and Auerbach's neuronal plexus. The findings of the present study confirm the homology of the clones isolated from the Colombian strain, with predominance of a 'principal clone' and an ubiquitous distribution of parasites belonging to a same clone. PMID- 16709230 TI - Heart failure alters MyoD and MRF4 expressions in rat skeletal muscle. AB - Heart failure (HF) is characterized by a skeletal muscle myopathy with increased expression of fast myosin heavy chains (MHCs). The skeletal muscle-specific molecular regulatory mechanisms controlling MHC expression during HF have not been described. Myogenic regulatory factors (MRFs), a family of transcriptional factors that control the expression of several skeletal muscle-specific genes, may be related to these alterations. This investigation was undertaken in order to examine potential relationships between MRF mRNA expression and MHC protein isoforms in Wistar rat skeletal muscle with monocrotaline-induced HF. We studied soleus (Sol) and extensor digitorum longus (EDL) muscles from both HF and control Wistar rats. MyoD, myogenin and MRF4 contents were determined using reverse transcription-polymerase chain reaction while MHC isoforms were separated using polyacrylamide gel electrophoresis. Despite no change in MHC composition of Wistar rat skeletal muscles with HF, the mRNA relative expression of MyoD in Sol and EDL muscles and that of MRF4 in Sol muscle were significantly reduced, whereas myogenin was not changed in both muscles. This down-regulation in the mRNA relative expression of MRF4 in Sol was associated with atrophy in response to HF while these alterations were not present in EDL muscle. Taken together, our results show a potential role for MRFs in skeletal muscle myopathy during HF. PMID- 16709231 TI - Developmental abnormalities in multiple proliferative tissues of Apc(Min/+) mice. AB - Germ-line mutation of the Apc gene has been linked to familial adenomatous polyposis (FAP) that predisposes to colon cancer. Apc(Min/+) mice, heterozygous for the Apc gene mutation, progressively develop small intestinal tumours in a manner that is analogous to that observed in the colon of patients with FAP (Su et al. 1992; Fodde et al. 1994; Moser et al. 1995). We have studied the effects of Apc gene mutation on murine intestinal and extra-intestinal, proliferatively active tissues. We have contrasted the histology to that of the age- and sex matched wild-type C57BL/6 mice. Histological assessment of the normal appearing intestinal mucosa demonstrates minimal change in size of crypts. In contrast, villi are longer in the ileum of Apc(Min/+) mice relative to C57BL/6 mice at 12 and 15 weeks of age. Vigorous splenic haematopoiesis in Apc(Min/+) mice was seen at 12 and 15 weeks of age, as reflected by marked splenomegaly, increased splenic haematopoietic cells and megakaryocytes. Peripheral blood counts, however, did not differ between C57BL/6 and Apc(Min/+) mice at 15 weeks of age. Lymphoid depletion in Apc(Min/+) mice was characterized by diminished numbers of splenic lymphoid follicles and small intestinal Peyer's patches. The ovaries of 12- and 15-week-old Apc(Min/+) mice exhibited increased numbers of atretic follicles, and estrous cycling by serial vaginal smears showed tendency of elongation in the mutant mice during these age ranges. The testicles of 10-week-old Apc(Min/+) mice showed increased numbers of underdeveloped seminiferous tubules. Collectively, these data suggest that, in addition to its obvious effects upon intestinal adenoma formation, Apc gene mutation causes impairment of developmental and apparent differentiation blockade in proliferative tissues, including those of the haematopoietic system, lymphoid and reproductive tract. PMID- 16709234 TI - Abstracts for the 20th EFI (European Federation for Immunogenetics) Conference, Oslo, Norway, 8-11 June, 2006. PMID- 16709232 TI - Morphological evaluation of the radioprotective effects of melatonin against X ray-induced early and acute testis damage in Albino rats: an animal model. AB - Irradiation has profound effects on the reproductive function. Our knowledge about radioprotective effects of melatonin against X-ray-induced testis damage is rudimentary. In this investigation, we hypothesized that melatonin can minimize germ-cell depletion and morphological features of cell damage in testis following X-ray irradiation (XRI). To examine these effects, and to test our hypothesis, an animal model comprised of 60 Albino rats was established. The animals were divided into five groups: Group 1, non-irradiated; Group 2, X-ray irradiated (XRI, 8 Grays); Group 3, XRI pretreated with solvent (ethanol and phosphate buffered saline); Group 4, non-irradiated group treated with melatonin and Group 5, XRI pretreated with melatonin. The testes were evaluated for both histological (light microscopy) and ultrastructural changes (transmission electron microscopy). Histologically, there were marked depletions (66%) of the germinal epithelial cells, in XRI group (Groups 2 and 3), whereas these changes were almost absent in XRI testis of animals pretreated with melatonin (Group 5). The number of spermatogenic cells in XRI testis of animals pretreated with melatonin (Group 5) was comparable (95%) to that of non-irradiated group (Groups 1 and 4) but significantly (P < 0.05) higher than those in XRI testis (34%, Groups 2 and 3). Ultrastructurally, XRI testis (Groups 2 and 3) showed features of apoptosis (condensation of the nuclei, vacuolization of the cytoplasm, increased cytoplasmic density and apoptotic bodies). These features were absent in XRI testis of animals pretreated with melatonin (Group 5). Also, this Group showed features of an increased metabolic activity (large acrosomal vesicle, prominent Golgi, increased mitotic activity, increased complement of cytoplasmic organelles and appearance of nucleoli-like bodies). There was a minimal depletion of the Sertoli and Leydig cells following XRI. Also, morphological features of apoptosis were infrequent in these cells. Administration of melatonin (MEL) prior to irradiation can protect testis against its destructive effects. The protective effects include amelioration of germ-cell depletion and apoptotic changes. The clinical ramifications of these observations mandate further studies. PMID- 16709240 TI - Use of a multi-way method to analyze the amino acid composition of a conserved group of orthologous proteins in prokaryotes. AB - BACKGROUND: Amino acids in proteins are not used equally. Some of the differences in the amino acid composition of proteins are between species (mainly due to nucleotide composition and lifestyle) and some are between proteins from the same species (related to protein function, expression or subcellular localization, for example). As several factors contribute to the different amino acid usage in proteins, it is difficult both to analyze these differences and to separate the contributions made by each factor. RESULTS: Using a multi-way method called Tucker3, we have analyzed the amino composition of a set of 64 orthologous groups of proteins present in 62 archaea and bacteria. This dataset corresponds to essential proteins such as ribosomal proteins, tRNA synthetases and translational initiation or elongation factors, which are common to all the species analyzed. The Tucker3 model can be used to study the amino acid variability within and between species by taking into consideration the tridimensionality of the data set. We found that the main factor behind the amino acid composition of proteins is independent of the organism or protein function analyzed. This factor must be related to the biochemical characteristics of each amino acid. The difference between the non-ribosomal proteins and the ribosomal proteins (which are rich in arginine and lysine) is the main factor behind the differences in amino acid composition within species, while G+C content and optimal growth temperature are the main factors behind the differences in amino acid usage between species. CONCLUSION: We show that a multi-way method is useful for comparing the amino acid composition of several groups of orthologous proteins from the same group of species. This kind of dataset is extremely useful for detecting differences between and within species. PMID- 16709241 TI - Cellular response to 5-fluorouracil (5-FU) in 5-FU-resistant colon cancer cell lines during treatment and recovery. AB - BACKGROUND: Treatment of cells with the anti-cancer drug 5-fluorouracil (5-FU) causes DNA damage, which in turn affects cell proliferation and survival. Two stable wild-type TP53 5-FU-resistant cell lines, ContinB and ContinD, generated from the HCT116 colon cancer cell line, demonstrate moderate and strong resistance to 5-FU, respectively, markedly-reduced levels of 5-FU-induced apoptosis, and alterations in expression levels of a number of key cell cycle- and apoptosis-regulatory genes as a result of resistance development. The aim of the present study was to determine potential differential responses to 8 and 24 hour 5-FU treatment in these resistant cell lines. We assessed levels of 5-FU uptake into DNA, cell cycle effects and apoptosis induction throughout treatment and recovery periods for each cell line, and alterations in expression levels of DNA damage response-, cell cycle- and apoptosis-regulatory genes in response to short-term drug exposure. RESULTS: 5-FU treatment for 24 hours resulted in S phase arrests, p53 accumulation, up-regulation of p53-target genes on DNA damage response (ATF3, GADD34, GADD45A, PCNA), cell cycle-regulatory (CDKN1A), and apoptosis-regulatory pathways (FAS), and apoptosis induction in the parental and resistant cell lines. Levels of 5-FU incorporation into DNA were similar for the cell lines. The pattern of cell cycle progression during recovery demonstrated consistently that the 5-FU-resistant cell lines had the smallest S phase fractions and the largest G2(/M) fractions. The strongly 5-FU-resistant ContinD cell line had the smallest S phase arrests, the lowest CDKN1A levels, and the lowest levels of 5-FU-induced apoptosis throughout the treatment and recovery periods, and the fastest recovery of exponential growth (10 days) compared to the other two cell lines. The moderately 5-FU-resistant ContinB cell line had comparatively lower apoptotic levels than the parental cells during treatment and recovery periods and a recovery time of 22 days. Mitotic activity ceased in response to drug treatment for all cell lines, consistent with down-regulation of mitosis-regulatory genes. Differential expression in response to 5-FU treatment was demonstrated for genes involved in regulation of nucleotide binding/metabolism (ATAD2, GNL2, GNL3, MATR3), amino acid metabolism (AHCY, GSS, IVD, OAT), cytoskeleton organization (KRT7, KRT8, KRT19, MAST1), transport (MTCH1, NCBP1, SNAPAP, VPS52), and oxygen metabolism (COX5A, COX7C). CONCLUSION: Our gene expression data suggest that altered regulation of nucleotide metabolism, amino acid metabolism, cytoskeleton organization, transport, and oxygen metabolism may underlie the differential resistance to 5-FU seen in these cell lines. The contributory roles to 5-FU resistance of some of the affected genes on these pathways will be assessed in future studies. PMID- 16709243 TI - A controlled trial of the effectiveness of a diabetes education programme in a multi-ethnic community in Glasgow [ISRCTN28317455]. AB - BACKGROUND: Epidemiologic data have shown that the prevalence of Type 2 diabetes varies with ethnic origin. Type 2 diabetes is up to four times more common in British South Asians than in the indigenous white population. The aim of this study was to develop a culturally appropriate educational intervention programme for South Asians with Type 2 diabetes. We then investigated whether this intervention could produce an improvement, and finally whether any improvement was greater than background changes in knowledge in comparison groups. METHODS: A multi-site prospective, randomised controlled study was conducted in all day care centres and three general practice registers with high proportion patients from different ethnic minority groups in Glasgow, Scotland. The intervention consisted of 18 educational sessions in 6 separate programmes. A modified questionnaire was used to measure the knowledge, attitudes, and practice of diabetes before and after intervention. RESULTS: Baseline assessment showed that Indian and Pakistani subjects had less knowledge about diabetes, regarded the disease less seriously, and had a lesser understanding of the relationship between control and complications than the white population. No differences in initial responses were found between those who completed the second assessment and those who did not. The intervention group showed significant improvements in scores for Knowledge (+12.5%); Attitudes toward seriousness (+13.5%), complications (+8.1%), Practice (+20.0%). However there were also changes in the ethnic control group scores; respectively +5.0%, +16.3% (significant P < 0.001), +1.5%, +1.7%. The single white control group also showed some improvements; respectively +12.2%, +12.4% (P = 0.04), +6.0%, +25.0% (P = 0.007), but the differences in improvement between these two control groups were not significant. Overall, the improvement seen was similar in both intervention and ethnic control groups and there was no significant difference in the amount of change (P = 0.36 CI -0.9 to +2.6). CONCLUSION: This study has shown that conducting a culturally-competent educational intervention in patients with Type 2 diabetes from ethnic minority groups is feasible and can improve their knowledge and attitudes and practice. However there was no net benefit compared with the control group. PMID- 16709242 TI - Microarray-based identification of antigenic variants of foot-and-mouth disease virus: a bioinformatics quality assessment. AB - BACKGROUND: The evolution of viral quasispecies can influence viral pathogenesis and the response to antiviral treatments. Mutant clouds in infected organisms represent the first stage in the genetic and antigenic diversification of RNA viruses, such as foot and mouth disease virus (FMDV), an important animal pathogen. Antigenic variants of FMDV have been classically diagnosed by immunological or RT-PCR-based methods. DNA microarrays are becoming increasingly useful for the analysis of gene expression and single nucleotide polymorphisms (SNPs). Recently, a FMDV microarray was described to detect simultaneously the seven FMDV serotypes. These results encourage the development of new oligonucleotide microarrays to probe the fine genetic and antigenic composition of FMDV for diagnosis, vaccine design, and to gain insight into the molecular epidemiology of this pathogen. RESULTS: A FMDV microarray was designed and optimized to detect SNPs at a major antigenic site of the virus. A screening of point mutants of the genomic region encoding antigenic site A of FMDV C-S8c1 was achieved. The hybridization pattern of a mutant includes specific positive and negative signals as well as crosshybridization signals, which are of different intensity depending on the thermodynamic stability of each probe-target pair. Moreover, an array bioinformatic classification method was developed to evaluate the hybridization signals. This statistical analysis shows that the procedure allows a very accurate classification per variant genome. CONCLUSION: A specific approach based on a microarray platform aimed at distinguishing point mutants within an important determinant of antigenicity and host cell tropism, namely the G-H loop of capsid protein VP1, was developed. The procedure is of general applicability as a test for specificity and discriminatory power of microarray based diagnostic procedures using multiple oligonucleotide probes. PMID- 16709244 TI - An active form of Vav1 induces migration of mammary epithelial cells by stimulating secretion of an epidermal growth factor receptor ligand. AB - BACKGROUND: Vav proteins are guanine nucleotide exchange factors (GEF) for Rho family GTPases and are activated following engagement of membrane receptors. Overexpression of Vav proteins enhances lamellipodium and ruffle formation, migration, and cell spreading, and augments activation of many downstream signaling proteins like Rac, ERK and Akt. Vav proteins are composed of multiple structural domains that mediate their GEF function and binding interactions with many cellular proteins. In this report we examine the mechanisms responsible for stimulation of cell migration by an activated variant of Vav1 and identify the domains of Vav1 required for this activity. RESULTS: We found that expression of an active form of Vav1, Vav1Y3F, in MCF-10A mammary epithelial cells increases cell migration in the absence or presence of EGF. Vav1Y3F was also able to drive Rac1 activation and PAK and ERK phosphorylation in MCF-10A cells in the absence of EGF stimulation. Mutations in the Dbl homology, pleckstrin homology, or cysteine-rich domains of Vav1Y3F abolished Rac1 or ERK activation in the absence of EGF and blocked the migration-promoting activity of Vav1Y3F. In contrast, mutations in the SH2 and C-SH3 domains did not affect Rac activation by Vav1Y3F, but reduced the ability of Vav1Y3F to induce EGF-independent migration and constitutive ERK phosphorylation. EGF-independent migration of MCF-10A cells expressing Vav1Y3F was abolished by treatment of cells with an antibody that prevents ligand binding to the EGF receptor. In addition, conditioned media collected from Vav1Y3F expressing cells stimulated migration of parental MCF-10A cells. Lastly, treatment of cells with the EGF receptor inhibitory antibody blocked the Vav1Y3F-induced, EGF-independent stimulation of ERK phosphorylation, but had no effect on Rac1 activation or PAK phosphorylation. CONCLUSION: Our results indicate that increased migration of active Vav1 expressing cells is dependent on Vav1 GEF activity and secretion of an EGF receptor ligand. In addition, activation of ERK downstream of Vav1 is dependent on autocrine EGF receptor stimulation while active Vav1 can stimulate Rac1 and PAK activation independent of ligand binding to the EGF receptor. Thus, stimulation of migration by activated Vav1 involves both EGF receptor-dependent and independent activities induced through the Rho GEF domain of Vav1. PMID- 16709246 TI - The continuing HIV vaccine saga: is a paradigm shift necessary? AB - As pointed out in previous editorials, the development of an effective vaccine for the Human Immunodeficiency Virus capable of preventing infection, or even one capable of preventing the Acquired Immunodeficiency Disease Syndrome, has eluded investigators for the past 20 years. Now Reche and Keskin and their co-workers have provided evidence that an entirely new approach, based upon modern bioinformatics methods and skillful in vitro immunological experiments, may result in an effective way to prime the T cell immune response of normal individuals against conserved peptide epitopes. PMID- 16709245 TI - Treatment of primary glioblastoma multiforme with cetuximab, radiotherapy and temozolomide (GERT)--phase I/II trial: study protocol. AB - BACKGROUND: The implementation of combined radiochemotherapy (RCHT) with temozolomide (TMZ) has lead to a significant increase in overall survival times in patients with Glioblastoma multiforme (GBM), however, outcome still remains unsatisfactory. The majority of GBMs show an overexpression and/or amplification of the epidermal growth factor receptor (EGFR). Therefore, addition of EGFR inhibition with cetuximab to the current standard treatment approach with radiotherapy and TMZ seems promising. METHODS/DESIGN: GERT is a one-armed single center phase I/II trial. In a first step, dose-escalation of TMZ from 50 mg/m2 to 75 mg/m2 together with radiotherapy and cetuximab will be performed. Should safety be proven, the phase II trial will be initiated with the standard dose of 75 mg/m2 of TMZ. Cetuximab will be applied in the standard application dose of 400 mg/m2 in week 1, thereafter at a dose of 250 mg/m2 weekly. A total of 46 patients will be included into this phase I/II trial. Primary endpoints are feasibility and toxicity, secondary endpoints are overall and progression-free survival. An interim analysis will be performed after inclusion of 15 patients into the main study. Patients' enrollment will be performed over a period of 2 years. The observation time will end 2 years after inclusion of the last patient. DISCUSSION: The goal of this study is to evaluate the safety and efficacy of combined RCHT-immunotherapy with TMZ and cetuximab as first-line treatment for patients with primary GBM. PMID- 16709247 TI - Variation in prescribing for anxiety and depression: a reflection of health inequalities, cultural differences or variations in access to care? AB - BACKGROUND: There are large variations in mental health prescribing in UK populations. However the underlying reasons for these differences, which may be related to differences in prevalence, cultural expectations or practical difficulties in access to treatment, remain uncertain. METHODS: Linear modelling was used to investigate whether population characteristics or access to primary care account for variations in mental health prescribing across 39 deprived neighbourhoods. RESULTS: The proportion of sampled respondents whose first language was not English and the ratio of general practitioners to population explained 61% of variation. Deprivation and mental health status were not significant predictors of prescribing in these relatively deprived communities. CONCLUSION: These findings suggest that mental health prescribing, within deprived areas, as well as reflecting cultural and social differences in prescribing, may also be a proxy measure of access to care. PMID- 16709248 TI - Haplotype-based quantitative trait mapping using a clustering algorithm. AB - BACKGROUND: With the availability of large-scale, high-density single-nucleotide polymorphism (SNP) markers, substantial effort has been made in identifying disease-causing genes using linkage disequilibrium (LD) mapping by haplotype analysis of unrelated individuals. In addition to complex diseases, many continuously distributed quantitative traits are of primary clinical and health significance. However the development of association mapping methods using unrelated individuals for quantitative traits has received relatively less attention. RESULTS: We recently developed an association mapping method for complex diseases by mining the sharing of haplotype segments (i.e., phased genotype pairs) in affected individuals that are rarely present in normal individuals. In this paper, we extend our previous work to address the problem of quantitative trait mapping from unrelated individuals. The method is non parametric in nature, and statistical significance can be obtained by a permutation test. It can also be incorporated into the one-way ANCOVA (analysis of covariance) framework so that other factors and covariates can be easily incorporated. The effectiveness of the approach is demonstrated by extensive experimental studies using both simulated and real data sets. The results show that our haplotype-based approach is more robust than two statistical methods based on single markers: a single SNP association test (SSA) and the Mann-Whitney U-test (MWU). The algorithm has been incorporated into our existing software package called HapMiner, which is available from our website at http://www.eecs.case.edu/~jxl175/HapMiner.html. CONCLUSION: For QTL (quantitative trait loci) fine mapping, to identify QTNs (quantitative trait nucleotides) with realistic effects (the contribution of each QTN less than 10% of total variance of the trait), large samples sizes (>or= 500) are needed for all the methods. The overall performance of HapMiner is better than that of the other two methods. Its effectiveness further depends on other factors such as recombination rates and the density of typed SNPs. Haplotype-based methods might provide higher power than methods based on a single SNP when using tag SNPs selected from a small number of samples or some other sources (such as HapMap data). Rank-based statistics usually have much lower power, as shown in our study. PMID- 16709249 TI - Linkage disequilibrium blocks, haplotype structure, and htSNPs of human CYP7A1 gene. AB - BACKGROUND: Cholesterol 7-alpha-hydroxylase (CYP7A1) is the rate limiting enzyme for converting cholesterol into bile acids. Genetic variations in the CYP7A1 gene have been associated with metabolic disorders of cholesterol and bile acids, including hypercholesterolemia, hypertriglyceridemia, arteriosclerosis, and gallstone disease. Current genetic studies are focused mainly on analysis of a single nucleotide polymorphism (SNP) at A-278C in the promoter region of the CYP7A1 gene. Here we report a genetic approach for an extensive analysis on linkage disequilibrium (LD) blocks and haplotype structures of the entire CYP7A1 gene and its surrounding sequences in Africans, Caucasians, Asians, Mexican Americans, and African-Americans. RESULT: The LD patterns and haplotype blocks of CYP7A1 gene were defined in Africans, Caucasians, and Asians using genotyping data downloaded from the HapMap database to select a set of haplotype-tagging SNPs (htSNP). A low cost, microarray-based platform on thin-film biosensor chips was then developed for high-throughput genotyping to study transferability of the HapMap htSNPs to Mexican-American and African-American populations. Comparative LD patterns and haplotype block structure was defined across all test populations. CONCLUSION: A constant genetic structure in CYP7A1 gene and its surrounding sequences was found that may lead to a better design for association studies of genetic variations in CYP7A1 gene with cholesterol and bile acid metabolism. PMID- 16709250 TI - Gene expression profiling of lymphoblastoid cell lines from monozygotic twins discordant in severity of autism reveals differential regulation of neurologically relevant genes. AB - BACKGROUND: The autism spectrum encompasses a set of complex multigenic developmental disorders that severely impact the development of language, non verbal communication, and social skills, and are associated with odd, stereotyped, repetitive behavior and restricted interests. To date, diagnosis of these neurologically based disorders relies predominantly upon behavioral observations often prompted by delayed speech or aberrant behavior, and there are no known genes that can serve as definitive biomarkers for the disorders. RESULTS: Here we demonstrate, for the first time, that lymphoblastoid cell lines from monozygotic twins discordant with respect to severity of autism and/or language impairment exhibit differential gene expression patterns on DNA microarrays. Furthermore, we show that genes important to the development, structure, and/or function of the nervous system are among the most differentially expressed genes, and that many of these genes map closely in silico to chromosomal regions containing previously reported autism candidate genes or quantitative trait loci. CONCLUSION: Our results provide evidence that novel candidate genes for autism may be differentially expressed in lymphoid cell lines from individuals with autism spectrum disorders. This finding further suggests the possibility of developing a molecular screen for autism based on expressed biomarkers in peripheral blood lymphocytes, an easily accessible tissue. In addition, gene networks are identified that may play a role in the pathophysiology of autism. PMID- 16709251 TI - Caloric restriction in C57BL/6J mice mimics therapeutic fasting in humans. AB - BACKGROUND: Caloric restriction (CR) has long been recognized as a dietary therapy that improves health and increases longevity. Little is known about the persistent effects of CR on plasma biomarkers (glucose, ketone bodies, and lipids) following re-feeding in mice. It is also unclear how these biomarker changes in calorically restricted mice relate to those observed previously in calorically restricted humans. RESULTS: Three groups of individually housed adult female C57BL/6J (B6) mice (n = 4/group) were fed a standard rodent chow diet either: (1) unrestricted (UR); (2) restricted for three weeks to reduce body weight by approximately 15-20% (R); or (3) restricted for three weeks and then re fed unrestricted (ad libitum) for an additional three weeks (R-RF). Body weight and food intake were measured throughout the study, while plasma lipids and levels of glucose and ketone bodies (beta-hydroxybutyrate) were measured at the termination of the study. Plasma glucose, phosphatidylcholine, cholesterol, and triglycerides were significantly lower in the R mice than in the UR mice. In contrast, plasma fatty acids and beta-hydroxybutyrate were significantly higher in the R mice than in the UR mice. CR had no effect on plasma phosphatidylinositol levels. While body weight and plasma lipids of the R-RF mice returned to unrestricted levels upon re-feeding, food intake and glucose levels remained significantly lower than those prior to the initiation of CR. CONCLUSION: CR establishes a new homeostatic state in B6 mice that persists for at least three weeks following ad libitum re-feeding. Moreover, the plasma biomarker changes observed in B6 mice during CR mimic those reported in humans on very low calorie diets or during therapeutic fasting. PMID- 16709252 TI - Capsaicin-resistant arterial baroreceptors. AB - BACKGROUND: Aortic baroreceptors (BRs) comprise a class of cranial afferents arising from major arteries closest to the heart whose axons form the aortic depressor nerve. BRs are mechanoreceptors that are largely devoted to cardiovascular autonomic reflexes. Such cranial afferents have either lightly myelinated (A-type) or non-myelinated (C-type) axons and share remarkable cellular similarities to spinal primary afferent neurons. Our goal was to test whether vanilloid receptor (TRPV1) agonists, capsaicin (CAP) and resiniferatoxin (RTX), altered the pressure-discharge properties of peripheral aortic BRs. RESULTS: Periaxonal application of 1 microM CAP decreased the amplitude of the C wave in the compound action potential conducting at <1 m/sec along the aortic depressor nerve. 10 microM CAP eliminated the C-wave while leaving intact the A wave conducting in the A-delta range (<12 m/sec). These whole nerve results suggest that TRPV1 receptors are expressed along the axons of C- but not A conducting BR axons. In an aortic arch--aortic nerve preparation, intralumenal perfusion with 1 microM CAP had no effect on the pressure-discharge relations of regularly discharging, single fiber BRs (A-type)--including the pressure threshold, sensitivity, frequency at threshold, or maximum discharge frequency (n = 8, p > 0.50) but completely inhibited discharge of an irregularly discharging BR (C-type). CAP at high concentrations (10-100 microM) depressed BR sensitivity in regularly discharging BRs, an effect attributed to non-specific actions. RTX (< or = 10 microM) did not affect the discharge properties of regularly discharging BRs (n = 7, p > 0.18). A CAP-sensitive BR had significantly lower discharge regularity expressed as the coefficient of variation than the CAP resistant fibers (p < 0.002). CONCLUSION: We conclude that functional TRPV1 channels are present in C-type but not A-type (A-delta) myelinated aortic arch BRs. CAP has nonspecific inhibitory actions that are unlikely to be related to TRV1 binding since such effects were absent with the highly specific TRPV1 agonist RTX. Thus, CAP must be used with caution at very high concentrations. PMID- 16709253 TI - Apoptosis of viral-infected airway epithelial cells limit viral production and is altered by corticosteroid exposure. AB - BACKGROUND: Effects of respiratory viral infection on airway epithelium include airway hyper-responsiveness and inflammation. Both features may contribute to the development of asthma. Excessive damage and loss of epithelial cells are characteristic in asthma and may result from viral infection. OBJECTIVE: To investigate apoptosis in Adenoviral-infected Guinea pigs and determine the role of death receptor and ligand expression in the airway epithelial response to limit viral infection. METHODS: Animal models included both an Acute and a Chronic Adeno-infection with ovalbumin-induced airway inflammation with/without corticosteroid treatment. Isolated airway epithelial cells were cultured to study viral production after infection under similar conditions. Immunohistochemistry, western blots and viral DNA detection were used to assess apoptosis, death receptor and TRAIL expression and viral release. RESULTS: In vivo and in vitro Adeno-infection demonstrated different apoptotic and death receptors (DR) 4 and 5 expression in response to corticosteroid exposure. In the Acute Adeno-infection model, apoptosis and DR4/5 expression was coordinated and were time-dependent. However, in vitro Acute viral infection in the presence of corticosteroids demonstrated delayed apoptosis and prolonged viral particle production. This reduction in apoptosis in Adeno-infected epithelial cells by corticosteroids exposure induced a prolonged virus production via both DR4 and TRAIL protein suppression. In the Chronic model where animals were ovalbumin sensitized/challenged and were treated with corticosteroids, apoptosis was reduced relative to adenovirus-infected or corticosteroid alone. CONCLUSION: Our data suggests that apoptosis of infected cells limits viral production and may be mediated by DR4/5 and TRAIL expression. In the Acute model of Adeno-infection, corticosteroid exposure may prolong viral particle production by altering this apoptotic response of the infected cells. This results from decreased DR4 and TRAIL expression. In the Chronic model treated with corticosteroids, a similar decreased apoptosis was observed. This data suggests that DR and TRAIL modulation by corticosteroids may be important in viral infection of airway epithelium. The prolonged virus release in the setting of corticosteroids may result from reduced apoptosis and suppressed DR4/TRAIL expression by the infected cells. PMID- 16709254 TI - The shortened disabilities of the arm, shoulder and hand questionnaire (QuickDASH): validity and reliability based on responses within the full-length DASH. AB - BACKGROUND: The 30-item disabilities of the arm, shoulder and hand (DASH) questionnaire is increasingly used in clinical research involving upper extremity musculoskeletal disorders. From the original DASH a shorter version, the 11-item QuickDASH, has been developed. Little is known about the discriminant ability of score changes for the QuickDASH compared to the DASH. The aim of this study was to assess the performance of the QuickDASH and its cross-sectional and longitudinal validity and reliability. METHODS: The study was based on extracting QuickDASH item responses from the responses to the full-length DASH questionnaire completed by 105 patients with a variety of upper extremity disorders before surgery and at follow-up 6 to 21 months after surgery. The DASH and QuickDASH scores were compared for the whole population and for different diagnostic groups. For longitudinal construct validity the effect size and standardized response mean were calculated. Analyses with ROC curves were performed to compare the ability of the DASH and QuickDASH to discriminate among patients classified according to the magnitude of self-rated improvement. Cross-sectional and test retest reliability was assessed. RESULTS: The mean DASH score was 34 (SD 22) and the mean QuickDASH score was 39 (SD 24) at baseline. For the different diagnostic groups the mean and median QuickDASH scores were higher than the corresponding DASH scores. For the whole population, the mean difference between the QuickDASH and DASH baseline scores was 4.2 (95% CI 3.2-5.3), follow-up scores was 2.6 (1.7 3.4), and change scores was 1.7 (0.6-2.8). The overall effect size and standardized response mean measured with the DASH and the QuickDASH were similar. In the ROC analysis of change scores among patients who rated their arm status as somewhat or much better and those who rated it as unchanged the difference in the area under the ROC curve for the DASH and QuickDASH was 0.01 (95% CI -0.05-0.07) indicating similar discriminant ability.Cross-sectional and test-retest reliability of the DASH and QuickDASH were similar. CONCLUSION: The results indicate that the QuickDASH can be used instead of the DASH with similar precision in upper extremity disorders. PMID- 16709256 TI - BAC-FISH refutes report of an 8p22-8p23.1 inversion or duplication in 8 patients with Kabuki syndrome. AB - BACKGROUND: Kabuki syndrome is a multiple congenital anomaly/mental retardation syndrome. The syndrome is characterized by varying degrees of mental retardation, postnatal growth retardation, distinct facial characteristics resembling the Kabuki actor's make-up, cleft or high-arched palate, brachydactyly, scoliosis, and persistence of finger pads. The multiple organ involvement suggests that this is a contiguous gene syndrome but no chromosomal anomalies have been isolated as an etiology. Recent studies have focused on possible duplications in the 8p22 8p23.1 region but no consensus has been reached. METHODS: We used bacterial artificial chromosome-fluorescent in-situ hybridization (BAC-FISH) and G-band analysis to study eight patients with Kabuki syndrome. RESULTS: Metaphase analysis revealed no deletions or duplications with any of the BAC probes. Interphase studies of the Kabuki patients yielded no evidence of inversions when using three-color FISH across the region. These results agree with other research groups' findings but disagree with the findings of Milunsky and Huang. CONCLUSION: It seems likely that Kabuki syndrome is not a contiguous gene syndrome of the 8p region studied. PMID- 16709255 TI - Procalcitonin is not sufficiently reliable to be the sole marker of neonatal sepsis of nosocomial origin. AB - BACKGROUND: It has recently been suggested that serum procalcitonin (PCT) is of value in the diagnosis of neonatal sepsis, with varying results. The aim of this prospective multicenter study was to assess the usefulness of PCT as a marker of neonatal sepsis of nosocomial origin. METHODS: One hundred infants aged between 4 and 28 days of life admitted to the Neonatology Services of 13 acute-care teaching hospitals in Spain over 1-year with clinical suspicion of neonatal sepsis of nosocomial origin were included in the study. Serum PCT concentrations were determined by a specific immunoluminometric assay. The reliability of PCT for the diagnosis of nosocomial neonatal sepsis at the time of suspicion of infection and at 12-24 h and 36-48 h after the onset of symptoms was calculated by receiver-operating characteristics (ROC) curves. The Youden's index (sensitivity + specificity - 1) was used for determination of optimal cutoff values of the diagnostic tests in the different postnatal periods. Sensitivity, specificity, and the likelihood ratio of a positive and negative result with the 95% confidence interval (CI) were calculated. RESULTS: The diagnosis of nosocomial sepsis was confirmed in 61 neonates. Serum PCT concentrations were significantly higher at initial suspicion and at 12-24 h and 36-48 h after the onset of symptoms in neonates with confirmed sepsis than in neonates with clinically suspected but not confirmed sepsis. Optimal PCT thresholds according to ROC curves were 0.59 ng/mL at the time of suspicion of sepsis (sensitivity 81.4%, specificity 80.6%); 1.34 ng/mL within 12-24 h of birth (sensitivity 73.7%, specificity 80.6%), and 0.69 ng/mL within 36-48 h of birth (sensitivity 86.5%, specificity 72.7%). CONCLUSION: Serum PCT concentrations showed a moderate diagnostic reliability for the detection of nosocomial neonatal sepsis from the time of suspicion of infection. PCT is not sufficiently reliable to be the sole marker of sepsis, but would be useful as part of a full sepsis evaluation. PMID- 16709257 TI - Microarray analysis identifies a set of CXCR3 and CCR2 ligand chemokines as early IFNbeta-responsive genes in peripheral blood lymphocytes in vitro: an implication for IFNbeta-related adverse effects in multiple sclerosis. AB - BACKGROUND: A substantial proportion of multiple sclerosis (MS) patients discontinue interferon-beta (IFNbeta) treatment due to various adverse effects, most of which emerge at the early phase after initiation of the treatment and then diminish with time. At present, the molecular mechanism underlying IFNbeta related adverse effects remains largely unknown. The aim of this study is to identify a comprehensive list of early IFNbeta-responsive genes (IRGs) in peripheral blood mononuclear cells (PBMC) that may play a key role in induction of adverse effects. METHODS: Total RNA of PBMC exposed to 50 ng/ml recombinant human IFNbeta for 3 to 24 hours in vitro was processed for cDNA microarray analysis, followed by quantitative real-time RT-PCR analysis. RESULTS: Among 1,258 genes on the array, IFNbeta elevated the expression of 107 and 87 genes, while it reduced the expression of 22 and 23 genes at 3 and 24 hours, respectively. Upregulated IRGs were categorized into conventional IFN-response markers, components of IFN-signaling pathways, chemokines, cytokines, growth factors, and their receptors, regulators of apoptosis, DNA damage, and cell cycle, heat shock proteins, and costimulatory and adhesion molecules. IFNbeta markedly upregulated CXCR3 ligand chemokines (SCYB11, SCYB10 and SCYB9) chiefly active on effector T helper type 1 (Th1) T cells, and CCR2 ligand chemokines (SCYA8 and SCYA2) effective on monocytes, whereas it downregulated CXCR2 ligand chemokines (SCYB2, SCYB1 and IL8) primarily active on neutrophils. CONCLUSION: IFNbeta immediately induces a burst of gene expression of proinflammatory chemokines in vitro that have potential relevance to IFNbeta-related early adverse effects in MS patients in vivo. PMID- 16709259 TI - The effects of multiple features of alternatively spliced exons on the K(A)/K(S) ratio test. AB - BACKGROUND: The evolution of alternatively spliced exons (ASEs) is of primary interest because these exons are suggested to be a major source of functional diversity of proteins. Many exon features have been suggested to affect the evolution of ASEs. However, previous studies have relied on the KA/KS ratio test without taking into consideration information sufficiency (i.e., exon length > 75 bp, cross-species divergence > 5%) of the studied exons, leading to potentially biased interpretations. Furthermore, which exon feature dominates the results of the KA/KS ratio test and whether multiple exon features have additive effects have remained unexplored. RESULTS: In this study, we collect two different datasets for analysis - the ASE dataset (which includes lineage-specific ASEs and conserved ASEs) and the ACE dataset (which includes only conserved ASEs). We first show that information sufficiency can significantly affect the interpretation of relationship between exons features and the KA/KS ratio test results. After discarding exons with insufficient information, we use a Boolean method to analyze the relationship between test results and four exon features (namely length, protein domain overlapping, inclusion level, and exonic splicing enhancer (ESE) frequency) for the ASE dataset. We demonstrate that length and protein domain overlapping are dominant factors, and they have similar impacts on test results of ASEs. In addition, despite the weak impacts of inclusion level and ESE motif frequency when considered individually, combination of these two factors still have minor additive effects on test results. However, the ACE dataset shows a slightly different result in that inclusion level has a marginally significant effect on test results. Lineage-specific ASEs may have contributed to the difference. Overall, in both ASEs and ACEs, protein domain overlapping is the most dominant exon feature while ESE frequency is the weakest one in affecting test results. CONCLUSION: The proposed method can easily find additive effects of individual or multiple factors on the KA/KS ratio test results of exons. Therefore, the system can analyze complex conditions in evolution where multiple features are involved. More factors can also be added into the system to extend the scope of evolutionary analysis of exons. In addition, our method may be useful when orthologous exons can not be found for the KA/KS ratio test. PMID- 16709258 TI - Lumbar segmental mobility disorders: comparison of two methods of defining abnormal displacement kinematics in a cohort of patients with non-specific mechanical low back pain. AB - BACKGROUND: Lumbar segmental rigidity (LSR) and lumbar segmental instability (LSI) are believed to be associated with low back pain (LBP), and identification of these disorders is believed to be useful for directing intervention choices. Previous studies have focussed on lumbar segmental rotation and translation, but have used widely varying methodologies. Cut-off points for the diagnosis of LSR & LSI are largely arbitrary. Prevalence of these lumbar segmental mobility disorders (LSMDs) in a non-surgical, primary care LBP population has not been established. METHODS: A cohort of 138 consecutive patients with recurrent or chronic low back pain (RCLBP) were recruited in this prospective, pragmatic, multi-centre study. Consenting patients completed pain and disability rating instruments, and were referred for flexion-extension radiographs. Sagittal angular rotation and sagittal translation of each lumbar spinal motion segment was measured from the radiographs, and compared to a reference range derived from a study of 30 asymptomatic volunteers. In order to define reference intervals for normal motion, and define LSR and LSI, we approached the kinematic data using two different models. The first model used a conventional Gaussian definition, with motion beyond two standard deviations (2sd) from the reference mean at each segment considered diagnostic of rotational LSMD and translational LSMD. The second model used a novel normalised within-subjects approach, based on mean normalised contribution-to-total-lumbar-motion. An LSMD was then defined as present in any segment that contributed motion beyond 2sd from the reference mean contribution-to-normalised-total-lumbar-motion. We described reference intervals for normal segmental mobility, prevalence of LSMDs under each model, and the association of LSMDs with pain and disability. RESULTS: With the exception of the conventional Gaussian definition of rotational LSI, LSMDs were found in statistically significant prevalences in patients with RCLBP. Prevalences at both the segmental and patient level were generally higher using the normalised within subjects model (2.8 to 16.8% of segments; 23.3 to 35.5% of individuals) compared to the conventional Gaussian model (0 to 15.8%; 4.7 to 19.6%). LSMDs are associated with presence of LBP, however LSMDs do not appear to be strongly associated with higher levels of pain or disability compared to other forms of non-specific LBP. CONCLUSION: LSMDs are a valid means of defining sub-groups within non-specific LBP, in a conservative care population of patients with RCLBP. Prevalence was higher using the normalised within-subjects contribution-to total-lumbar-motion approach. PMID- 16709266 TI - Abstracts of the 41st Annual Meeting of the Association for European Paediatric Cardiology, Basel, Switzerland, 24-27 May, 2006. PMID- 16709267 TI - Can heterotopic gastric mucosa present as globus pharyngeus? AB - We present a series of four patients presenting with symptoms leading to a provisional diagnosis of globus pharyngeus. On further investigation, they were all found to have small areas of abnormal mucosa in either their hypopharynx or proximal oesophagus, which on biopsy were reported as containing gastric-type mucosa. Following anti-reflux treatment, all patients eventually reported a cessation of symptoms. All but one patient was followed up with repeat endoscopies. We highlight the fact that biopsies reporting gastric epithelium at this anatomical site are in keeping with the diagnosis of heterotopic gastric mucosa. We review the literature and report the high incidence and natural history of this condition. Heterotopic gastric mucosa can only be diagnosed on endoscopy and is easily missed, even during such an investigation. We raise the question of whether this condition may be one of a number of factors in the aetiology of globus pharyngeus. PMID- 16709260 TI - Large-scale and high-confidence proteomic analysis of human seminal plasma. AB - BACKGROUND: The development of mass spectrometric (MS) techniques now allows the investigation of very complex protein mixtures ranging from subcellular structures to tissues. Body fluids are also popular targets of proteomic analysis because of their potential for biomarker discovery. Seminal plasma has not yet received much attention from the proteomics community but its characterization could provide a future reference for virtually all studies involving human sperm. The fluid is essential for the survival of spermatozoa and their successful journey through the female reproductive tract. RESULTS: Here we report the high confidence identification of 923 proteins in seminal fluid from a single individual. Fourier transform MS enabled parts per million mass accuracy, and two consecutive stages of MS fragmentation allowed confident identification of proteins even by single peptides. Analysis with GoMiner annotated two-thirds of the seminal fluid proteome and revealed a large number of extracellular proteins including many proteases. Other proteins originated from male accessory glands and have important roles in spermatozoan survival. CONCLUSION: This high confidence characterization of seminal plasma content provides an inventory of proteins with potential roles in fertilization. When combined with quantitative proteomics methodologies, it should be useful for studies of fertilization, male infertility, and prostatic and testicular cancers. PMID- 16709268 TI - Observational study of bone-anchored hearing aid infection rates using different post-operative dressings. AB - INTRODUCTION: Avoidance of infection at the implant site is a crucial element to the success of bone-anchored hearing aid (BAHA) implantation. However, little evidence exists to suggest the best post-operative wound dressing to use. MATERIAL AND METHODS: We report our experience with 160 consecutive BAHA patients, using four types of post-operative BAHA wound dressing (Tri-adcortyl, Mepitel, Allevyn, and Acticoat 7 with Allevyn). Patients were reviewed at week one and week two post-operatively. Infection was defined as a positive wound swab culture or evidence of cellulitis at the BAHA site. RESULTS: Post-operative BAHA infection rates were 16 per cent, 50 per cent, 10 per cent and 5 per cent, for each of the four dressings respectively, and the mean number of additional visits to wound care clinic were 1.5, 3.7, one and 0.4, respectively. DISCUSSION: Acticoat 7 with Allevyn produced the lowest infection rate and thus became the dressing of choice for our BAHA programme. PMID- 16709269 TI - Foreign body mimicking a nasal bone fracture. AB - Nasal foreign bodies are common in young children and are typically the result of intranasal placement by the child. The authors report a case of an extranasal foreign body in an adult, which presented as a nasal fracture following trauma to the nose. This uncommon presentation, previously unreported in the literature, highlights the importance of careful history taking in cases of nasal trauma and of thorough wound exploration if any penetrative injury is found. PMID- 16709270 TI - Tracheoinnominate fistula: surgical management of an iatrogenic disaster. AB - Tracheoinnominate fistula (TIF) is a rare condition with significant potential for mortality if surgical intervention is not immediate. We present two cases of successfully managed TIF. Both cases involve ligation and resection of the innominate artery at the TIF followed by a pectoralis major muscle flap. In both cases, success was largely due to a high index of suspicion and immediate control of the bleeding with transport to the operating room for surgical repair. The history, aetiology, and pathogenesis of TIF are reviewed, yielding an algorithm for recommended management of TIF. PMID- 16709271 TI - Surgical debridement of craniocervical necrotizing fasciitis: the window of opportunity. AB - Craniocervical necrotizing fasciitis (CCNF) has a potentially high morbidity and mortality. Late presentation, shock and disseminated intravascular coagulation are associated with a particularly poor prognosis. Early recognition and aggressive treatment is advised. A case report is presented of a late presentation of necrotizing fasciitis of the face, neck and anterior thoracic wall. Despite adverse prognostic indicators, surgical debridement was performed. Intensive medical treatment included continuous renal replacement therapy and intravenous immunoglobulin. Survival in this case illustrates that the window of opportunity for surgically treating CCNF extends to the advanced stages of the disease, albeit with increased morbidity. PMID- 16709272 TI - Comparative gene mapping in Arabidopsis lyrata chromosomes 1 and 2 and the corresponding A. thaliana chromosome 1: recombination rates, rearrangements and centromere location. AB - To add detail to the genetic map of Arabidopsis lyrata, and compare it with that of A. thaliana, we have developed many additional markers in the A. lyrata linkage groups, LG1 and LG2, corresponding to A. thaliana chromosome 1. We used a newly developed method for marker development for single nucleotide polymorphisms present in gene sequences, plus length differences, to map genes in an A. lyrata family, including variants in several genes close to the A. thaliana centromere 1, providing the first data on the location of an A. lyrata centromere; we discuss the implications for the evolution of chromosome 1 of A. thaliana. With our larger marker density, large rearrangements between the two Arabidopsis species are excluded, except for a large inversion on LG2. This was previously known in Capsella; its presence in A. lyrata suggests that, like most other rearrangements, it probably arose in the A. thaliana lineage. Knowing that marker orders are similar, we can now compare homologous, non-rearranged map distances to test the prediction of more frequent crossing-over in the more inbreeding species. Our results support the previous conclusion of similar distances in the two species for A. lyrata LG1 markers. For LG2 markers, the distances were consistently, but non-significantly, larger in A. lyrata. Given the two species' large DNA content difference, the similarity of map lengths, particularly for LG1, suggests that crossing-over is more frequent across comparable physical distances in the inbreeder, A. thaliana, as predicted. PMID- 16709273 TI - Genetic architecture of variation in sex-comb tooth number in Drosophila simulans. AB - The sex comb on the forelegs of Drosophila males is a secondary sexual trait, and the number of teeth on these combs varies greatly within and between species. To understand the relationship between the intra- and interspecific variation, we performed quantitative trait locus (QTL) analyses of the intraspecific variation in sex-comb tooth number. We used five mapping populations derived from two inbred Drosophila simulans strains that were divergent in the number of sex-comb teeth. Although no QTLs were detected on the X chromosome, we identified four QTLs on the second chromosome and three QTLs on the third chromosome. While identification and estimated effects of the second-chromosome QTLs depend on genetic backgrounds, significant and consistent effects of the two third chromosome QTLs were found in two genetic backgrounds. There were significant epistatic interactions between a second-chromosome QTL and a third-chromosome QTL, as well as between two second-chromosome QTLs. The third-chromosome QTLs are concordant with the locations of the QTLs responsible for the previously observed differences in sex-comb tooth number between D. simulans and D. mauritiana. PMID- 16709274 TI - Microsatellites reveal extensive geographical, ecological and genetic contacts between invasive and indigenous whitefly biotypes in an insular environment. AB - Human-mediated bioinvasions provide the opportunity to study the early stages of contact between formerly allopatric, divergent populations of a species. However, when invasive and resident populations are morphologically similar, it may be very difficult to assess their distribution in the field, as well as the extent of ecological overlap and genetic exchanges between invasive and resident populations. We here illustrate the use of data obtained from a set of eight microsatellite markers together with Bayesian clustering methods to document invasions in a group of major tropical pests, Bemisia tabaci, which comprises several morphologically indistinguishable biotypes with different agronomic impacts. We focus on the island of La Reunion, where an invasive biotype (B) has recently been introduced and now interacts with the resident biotype (Ms). The temporal and spatial distribution, host-plant range and genetic structure of both biotypes are investigated. We showed (i) that, without prior information, clustering methods separate two groups of individuals that can safely be identified as the B and Ms biotypes; (ii) that the B biotype has invaded all regions of the island, and showed no signs of genetic founder effect relative to the Ms biotype; (iii) that the B and Ms biotypes coexist in sympatry throughout most of their geographical ranges, although they tend to segregate into different host plants; and finally (iv) that asymmetrical and locus-specific introgression occurs between the two biotypes when they are in syntopy. PMID- 16709275 TI - Quantifying the genomic decay paradox due to Muller's ratchet in human mitochondrial DNA. AB - The observation of high mitochondrial mutation rates in human pedigrees has led to the question of how such an asexual genetic system can survive the accumulation of slightly deleterious mutations caused by Muller's ratchet. I define a null model to quantify in unprecedented detail the threat from extinction caused by Muller's ratchet. This model is general enough to explore the biological significance of Muller's ratchet in various species where its operation has been suspected. For increased precision over a wide range of parameter space I employ individual-based simulations run by evolution@home, the first global computing system for evolutionary biology. After compiling realistic values for the key parameters in human mitochondrial DNA (mtDNA) I find that a surprisingly large range of biologically realistic parameter combinations would lead to the extinction of the human line over a period of 20 million years - if accepted wisdom about mtDNA and Muller's ratchet is correct. The resulting genomic decay paradox complements a similar threat from extinction due to mutation accumulation in nuclear DNA and suggests evaluation of unconventional explanations for long-term persistence. A substantial list of potential solutions is given, including compensatory back mutations, mutation rate heterogeneity and occasional recombination in mtDNA. Future work will have to explore which of these actually solves the paradox. Nonetheless, the results presented here provide yet another reason to minimize anthropogenic increase of mutation rates. PMID- 16709277 TI - K-means clustering: a half-century synthesis. AB - This paper synthesizes the results, methodology, and research conducted concerning the K-means clustering method over the last fifty years. The K-means method is first introduced, various formulations of the minimum variance loss function and alternative loss functions within the same class are outlined, and different methods of choosing the number of clusters and initialization, variable preprocessing, and data reduction schemes are discussed. Theoretic statistical results are provided and various extensions of K-means using different metrics or modifications of the original algorithm are given, leading to a unifying treatment of K-means and some of its extensions. Finally, several future studies are outlined that could enhance the understanding of numerous subtleties affecting the performance of the K-means method. PMID- 16709278 TI - Bootstrap scree tests: a Monte Carlo simulation and applications to published data. AB - A non-parametric procedure for Cattell's scree test is proposed, using the bootstrap method. Bentler and Yuan developed parametric tests for the linear trend of scree eigenvalues in principal component analysis. The proposed method is for cases where parametric assumptions are not realistic. We define the break in the scree trend in several ways, based on linear slopes defined with two or three consecutive eigenvalues, or all eigenvalues after the k largest. The resulting scree test statistics are evaluated under various data conditions, among which Gorsuch and Nelson's bootstrap CNG performs best and is reasonably consistent and efficient under leptokurtic and skewed conditions. We also examine the bias-corrected and accelerated bootstrap method for these statistics, and the bias correction is found to be too unstable to be useful. Using seven published data sets which Bentler and Yuan analysed, we compare the bootstrap approach to the scree test with the parametric linear trend test. PMID- 16709279 TI - The use of configural frequency analysis for explorative data analysis. AB - Configural frequency analysis (CFA) is a widely used method of explorative data analysis. It tries to detect patterns in the data that occur significantly more or significantly less often than expected by chance. Patterns which occur more often than expected by chance are called CFA types, while those which occur less often than expected by chance are called CFA antitypes. The patterns detected are used to generate knowledge about the mechanisms underlying the data. We investigate the ability of CFA to detect adequate types and antitypes in a number of simulation studies. The basic idea of these studies is to predefine sets of types and antitypes and a mechanism which uses them to create a simulated data set. This simulated data set is then analysed with CFA and the detected types and antitypes are compared to the predefined ones. The predefined types and antitypes together with the method to generate the data are called a data generation model. The results of the simulation studies show that CFA can be used in quite different research contexts to detect structural dependencies in observed data. In addition, we can learn from these simulation studies how much data is necessary to enable CFA to reconstruct the predefined types and antitypes with sufficient accuracy. For one of the data generation models investigated, implicitly underlying knowledge space theory, it was shown that zero-order CFA can be used to reconstruct the predefined types (which can be interpreted in this context as knowledge states) with sufficient accuracy. Theoretical considerations show that first-order CFA cannot be used for this data generation model. Thus, it is wrong to consider first-order CFA, as is done in many publications, as the standard or even only method of CFA. PMID- 16709280 TI - Maximal reliability and power in covariance structure models. AB - In covariance structure modelling, the non-centrality parameter of the asymptotic chi-squared distribution is typically used as an indicator of asymptotic power for hypothesis tests. When a latent linear regression is of interest, the contribution to power by the maximal reliability coefficient, which is associated with used latent variable indicators, is examined and this relationship is further explicated in the case of congeneric measures. It is also shown that item parcelling may reduce power of tests of latent regression parameters. Recommendations on weights for parcelling to avoid power loss are provided, which are found to be those of optimal linear composites with maximal reliability. PMID- 16709281 TI - Interval estimates of weighted effect sizes in the one-way heteroscedastic ANOVA. AB - A framework for comparing normal population means in the presence of heteroscedasticity and outliers is provided. A single number called the weighted effect size summarizes the differences in population means after weighting each according to the difficulty of estimating their respective means, whether the difficulty is due to unknown population variances, unequal sample sizes or the presence of outliers. For an ANOVA weighted for unequal variances, we find interval estimates for the weighted effect size. In addition, the weighted effect size is shown to be a monotone function of a suitably defined weighted coefficient of determination, which means that interval estimates of the former are readily transformed into interval estimates of the latter. Extensive simulations demonstrate the accuracy of the nominal 95% coverage of these intervals for a wide range of parameters. PMID- 16709282 TI - Generalized reliability estimation using repeated measurements. AB - Reliability can be studied in a generalized way using repeated measurements. Linear mixed models are used to derive generalized test-retest reliability measures. The method allows for repeated measures with a different mean structure due to correction for covariate effects. Furthermore, different variance covariance structures between measurements can be implemented. When the variance structure reduces to a random intercept (compound symmetry), classical methods are recovered. With more complex variance structures (e.g. including random slopes of time and/or serial correlation), time-dependent reliability functions are obtained. The effect of time lag between measurements on reliability estimates can be evaluated. The methodology is applied to a psychiatric scale for schizophrenia. PMID- 16709283 TI - Selecting among three-mode principal component models of different types and complexities: a numerical convex hull based method. AB - Several three-mode principal component models can be considered for the modelling of three-way, three-mode data, including the Candecomp/Parafac, Tucker3, Tucker2, and Tucker1 models. The following question then may be raised: given a specific data set, which of these models should be selected, and at what complexity (i.e. with how many components)? We address this question by proposing a numerical model selection heuristic based on a convex hull. Simulation results show that this heuristic performs almost perfectly, except for Tucker3 data arrays with at least one small mode and a relatively large amount of error. PMID- 16709284 TI - Bayesian analysis of structural equation models with mixed exponential family and ordered categorical data. AB - Structural equation models are very popular for studying relationships among observed and latent variables. However, the existing theory and computer packages are developed mainly under the assumption of normality, and hence cannot be satisfactorily applied to non-normal and ordered categorical data that are common in behavioural, social and psychological research. In this paper, we develop a Bayesian approach to the analysis of structural equation models in which the manifest variables are ordered categorical and/or from an exponential family. In this framework, models with a mixture of binomial, ordered categorical and normal variables can be analysed. Bayesian estimates of the unknown parameters are obtained by a computational procedure that combines the Gibbs sampler and the Metropolis-Hastings algorithm. Some goodness-of-fit statistics are proposed to evaluate the fit of the posited model. The methodology is illustrated by results obtained from a simulation study and analysis of a real data set about non adherence of hypertension patients in a medical treatment scheme. PMID- 16709285 TI - Limited-information goodness-of-fit testing of item response theory models for sparse 2 tables. AB - Bartholomew and Leung proposed a limited-information goodness-of-fit test statistic (Y) for models fitted to sparse 2(P ) contingency tables. The null distribution of Y was approximated using a chi-squared distribution by matching moments. The moments were derived under the assumption that the model parameters were known in advance and it was conjectured that the approximation would also be appropriate when the parameters were to be estimated. Using maximum likelihood estimation of the two-parameter logistic item response theory model, we show that the effect of parameter estimation on the distribution of Y is too large to be ignored. Consequently, we derive the asymptotic moments of Y for maximum likelihood estimation. We show using a simulation study that when the null distribution of Y is approximated using moments that take into account the effect of estimation, Y becomes a very useful statistic to assess the overall goodness of fit of models fitted to sparse 2(P) tables. PMID- 16709286 TI - Analysing multitrait-multimethod data with structural equation models for ordinal variables applying the WLSMV estimator: what sample size is needed for valid results? AB - Convergent and discriminant validity of psychological constructs can best be examined in the framework of multitrait-multimethod (MTMM) analysis. To gain information at the level of single items, MTMM models for categorical variables have to be applied. The CTC(M-1) model is presented as an example of an MTMM model for ordinal variables. Based on an empirical application of the CTC(M-1) model, a complex simulation study was conducted to examine the sample size requirements of the robust weighted least squares mean- and variance-adjusted chi(2) test of model fit (WLSMV estimator) implemented in Mplus. In particular, the simulation study analysed the chi(2) approximation, the parameter estimation bias, the standard error bias, and the reliability of the WLSMV estimator depending on the varying number of items per trait-method unit (ranging from 2 to 8) and varying sample sizes (250, 500, 750, and 1000 observations). The results showed that the WLSMV estimator provided a good -- albeit slightly liberal -- chi(2) approximation and stable and reliable parameter estimates for models of reasonable complexity (2-4 items) and small sample sizes (at least 250 observations). When more complex models with 5 or more items were analysed, larger sample sizes of at least 500 observations were needed. The most complex model with 9 trait-method units and 8 items (72 observed variables) requires sample sizes of at least 1000 observations. PMID- 16709303 TI - Maintenance plus reliever budesonide/formoterol compared with a higher maintenance dose of budesonide/formoterol plus formoterol as reliever in asthma: an efficacy and cost-effectiveness study. AB - OBJECTIVE: To evaluate efficacy and cost-effectiveness of budesonide/formoterol (Symbicort) maintenance (one dose once or twice daily) plus additional doses as needed (Symbicort Maintenance And Reliever Therapy, SMART) compared with a higher fixed dose of budesonide/formoterol with formoterol as needed in patients with persistent asthma. STUDY DESIGN AND METHODS: 6-month, open, randomised study of 465 patients either not well controlled on an inhaled corticosteroid (ICS), or well controlled on a combination of ICS and a long-acting beta(2)-agonist (LABA). TREATMENTS: budesonide/formoterol 160/4.5 microg, one inhalation, once or twice daily maintenance plus additional doses as-needed (1 x SMART or 2 x SMART), or budesonide/formoterol 160/4.5 microg two inhalations twice daily plus formoterol 4.5 microg as needed (2 x 2 FIX + F). Children 6-11 years old used an 80/4.5 microg dose strength. Primary variables of efficacy were the changes in the Asthma Control Questionnaire (ACQ(5)) and morning peak expiratory flow (PEF). RESULTS: Mean age of patients 40 years (range 6-82 years); 53% female. No differences between the groups were found in ACQ(5) scores or asthma exacerbation rates. Morning PEF was higher in the 2 x 2 FIX + F group vs. the 1 x SMART and 2 x SMART groups (differences 13 L/min and 9 L/min, respectively; p < 0.002). The 1 x SMART group showed a significant decrease in asthma controlled days compared with the two other groups. No difference was seen between the 2 x SMART group and the 2 x 2 FIX + F group. Treatment costs were significantly lower in the SMART groups compared with the 2 x 2 FIX + F group. CONCLUSION: Compared with the 2 x 2 FIX + F treatment the use of budesonide/formoterol was 30-40% lower in the SMART groups while maintaining equal ACQ(5) scores. Daily asthma control improved equally with 2 x SMART compared to 2 x 2 FIX + F with a reduction in asthma medication cost. The one dose once daily maintenance treatment (1 x SMART) resulted in a low level of treatment failure (exacerbations) but led to more days with symptoms. Therefore, a daily dose of two inhalations seems to be the lowest appropriate dose in patients with moderate persistent asthma. PMID- 16709304 TI - Consistency of lipid-altering effects of ezetimibe/simvastatin across gender, race, age, baseline low density lipoprotein cholesterol levels, and coronary heart disease status: results of a pooled retrospective analysis. AB - BACKGROUND: The combination tablet containing ezetimibe and simvastatin (EZE/SIMVA), inhibits both the intestinal absorption and endogenous production of cholesterol, providing significantly greater low-density lipoprotein cholesterol (LDL-C) lowering than EZE or SIMVA alone. The purpose of this pooled analysis was to evaluate the consistency of efficacy (i.e., between-treatment difference) of EZE/SIMVA versus SIMVA within several selected subgroups of patients with primary hypercholesterolemia. METHODS: For the present analysis, data were pooled from three similarly designed, 12-week, randomized, double-blind, placebo-controlled factorial studies consisting of 3083 patients with primary hypercholesterolemia (n = 311 in placebo group; n = 302 in EZE group; n = 1234 in pooled SIMVA group; n = 1236 in pooled EZE/SIMVA group). In these clinical studies, primary hypercholesterolemia was defined as an LDL-C value between 145 and 250 mg/dL inclusive and a triglyceride (TG) level of less than 350 mg/dL. The results for the pooled SIMVA and pooled EZE/SIMVA groups were used for the present analyses. The pooled analyses focused on the consistency of the between-treatment differences (i.e., incremental effect) for EZE/SIMVA (pooled across doses) versus SIMVA (pooled across doses) on various lipid and non-lipid parameters within different patient subgroups defined according to gender, race (Caucasian, Non Caucasian), baseline age (< 65, > or = 65 years), baseline LDL-C (< 160, > or = 160 mg/dL), and coronary heart disease (CHD) history. Tolerability was also examined for pooled EZE/SIMVA and pooled SIMVA within these selected subgroups. In a modified intention-to-treat analysis, an ANOVA model was used for testing the consistency of pooled treatment effects on lipid and non-lipid parameters within each selected subgroup. RESULTS: For the entire cohort, baseline lipid profiles were similar for the patients in the pooled EZE/SIMVA group compared with those in the pooled SIMVA group. Treatment with EZE/SIMVA led to significant (p < 0.001) incremental improvements in LDL-C, non-high density lipoprotein cholesterol (non-HDL-C), apolipoprotein B, TG and high sensitivity C-reactive protein compared to SIMVA, across the entire cohort. These changes were consistent within each of the selected subgroups. Moreover, more patients attained LDL-C goal levels < 100 mg/dL with EZE/SIMVA than with SIMVA in the entire cohort and this was consistent across all subgroups, except baseline LDL C. In this pooled retrospective analysis, treatment with EZE/SIMVA was generally well tolerated across subgroups, with a safety profile similar to SIMVA monotherapy. Although this pooled analysis was performed on a large cohort of patients with primary hypercholesterolemia, the results of this analysis were specific for this select patient population and generalizations to other populations should be applied with caution. CONCLUSION: The enhanced lipid altering effects of EZE/SIMVA versus those of SIMVA observed in the entire cohort were consistent within all subgroups examined. EZE/SIMVA represents an effective and well-tolerated therapeutic option for the treatment of a wide range of patient subgroups with primary hypercholesterolemia. PMID- 16709305 TI - Dosing patterns, hematologic outcomes, and costs of erythropoietic agents in predialysis chronic kidney disease patients with anemia. AB - OBJECTIVE: Few observational studies have evaluated the use of epoetin alfa (EPO) and darbepoetin alfa (DARB) in chronic kidney disease (CKD) patients with anemia. The objective of this study was to investigate dosing patterns, hematologic outcomes, and intervention costs with EPO and DARB in anemic CKD patients treated in an ambulatory care setting. METHODS: This was a multicenter, retrospective, chart review of predialysis CKD patients with anemia treated with EPO or DARB. Charts were sequentially selected from 435 EPO and 432 DARB patients naive to erythropoietic therapy and treated for > or = 24 weeks. Hemoglobin (Hb) levels, dates, and EPO/DARB doses were recorded. Drug costs using 2005 wholesale acquisition costs (WAC) and Federal Supply Schedule (FSS) pricing were based on the mean cumulative drug dose over the 24-week study period. RESULTS: A total of 393 EPO and 396 DARB charts met all criteria with predominantly male subjects (EPO: 94%; DARB: 96%). Mean baseline GFR and Hb levels were similar. Once-weekly and extended dosing (> or = Q2W) was common in both groups. At Weeks 4, 8, and 12 following initiation of therapy, a greater proportion of EPO than DARB patients reached target Hb levels (> or = 11 g/dL) (p < 0.0001); at Week 24, all patients reached target Hb levels. Mean 24-week cumulative doses were EPO 279 336 +/- 68 302 units and DARB 1084 +/- 246 microg. Drug cost was higher for DARB independent of pricing utilized (WAC: EPO = 3400 US dollars, DARB = 4726 US dollars; FSS: EPO = 1528 US dollars, DARB = 2379 US dollars). CONCLUSIONS: Extended dosing (Q2W) was common in EPO- and DARB-treated patients with CKD-related anemia, with EPO treated patients experiencing a significantly greater hematologic response (at Weeks 4, 8, and 12). In addition, drug cost was 39-56% higher in the DARB group. The male predominance may limit generalizability, warranting further research in other populations. PMID- 16709306 TI - Pneumonia due to Pseudomonas aeruginosa: the levofloxacin clinical trials experience. AB - Respiratory infections caused by Pseudomonas aeruginosa present significant treatment challenges, including that of overcoming intrinsic and adaptive resistance by these organisms. The fluoroquinolones may provide an effective option for treating these infections. In this analysis, we report on the efficacy of levofloxacin in the treatment of community-acquired pneumonia (CAP) and nosocomial pneumonia caused by P. aeruginosa using information from nine clinical studies supported by Johnson & Johnson Pharmaceutical Research and Development (Raritan, NJ) or Ortho-McNeil Pharmaceutical (Raritan, NJ). From these studies, a total of 36 patients were identified with pneumonia caused by P. aeruginosa and treated with levofloxacin (750 mg or 500 mg). For patients diagnosed with nosocomial pneumonia, levofloxacin treatment achieved a 64.7% (11/17) clinical success rate, compared with 41.2% (7/17) with comparator treatment (imipenem/cilastatin followed by ciprofloxacin) in the microbiologically evaluable population. Eradication rates were 58.8% with levofloxacin treatment vs. 29.4% with comparator (95% CI, -64.2 to 5.4). For levofloxacin-treated CAP patients with P. aeruginosa infections (n = 19), clinical success and microbiological eradication rates in the microbiologically evaluable population were 89.5% and 78.9%, respectively. Several limitations of this analysis exist including that this was a retrospective evaluation that pooled data from multiple studies with varying protocols, the number of patients included was limited, and the nosocomial pneumonia patients used adjunctive therapy with an antipseudomonal beta-lactam in most cases. Nonetheless, these findings suggest that levofloxacin may play a role in the treatment of these difficult respiratory infections. PMID- 16709307 TI - Improving the global management of the neurogenic bladder patient: part II. Future treatment strategies. AB - BACKGROUND: Patients with neurogenic bladder represent a small fraction of the total overactive bladder population. As a consequence, development of new therapies in this area has largely focused on idiopathic urinary incontinence. The absence of data for patients with neurological disease has far-reaching implications, affecting reimbursement and physicians' willingness to prescribe therapies, and limiting access of potential valuable treatments to patients whose lives are significantly impaired by inadequately managed bladder symptoms. SCOPE: The range of new therapies is increasing. Although many reviews of the overall safety, efficacy and mode of action of such treatments are available, there is limited information on how these treatments will best be used in clinical practice. We considered the current benefits and limitations of the various new licensed and unlicensed therapies and what role each would have in the future management of neurogenic urinary incontinence. CONCLUSIONS: A wide range of new treatments have been investigated for the management of overactive bladder; few, however, have been evaluated extensively in neurogenic urinary incontinence. Further studies are required to determine the optimal dosing regimes and formulations for individual sub-populations of neurogenic bladder patients and to determine the cost-effectiveness of these interventions. With the current experience available, two treatment algorithms for a subset of patients with neurological disease have also been proposed, which suggest at which stage of management and in which patients individual therapies for neurogenic urinary incontinence could be used. PMID- 16709308 TI - Cost-utility analysis in a UK setting of self-monitoring of blood glucose in patients with type 2 diabetes. AB - BACKGROUND: Self-monitoring of blood glucose (SMBG) in type 2 diabetes patients has been shown in meta-analyses of randomized trials to improve HbA(1c) by approximately 0.4% when compared to no SMBG. However, the cost of testing supplies is high, improvements in health utility due to improved glycaemic control may be possible and cost-effectiveness has not been evaluated. METHODS: A peer-reviewed validated model projected improvements in lifetime quality-adjusted life years (QALYs), long-term costs and cost-effectiveness of SMBG versus no SMBG. Markov/Monte Carlo modelling simulated the progression of complications (cardiovascular, neuropathy, renal and eye disease). Transition probabilities and HbA(1c)-dependent adjustments came from the United Kingdom Prospective Diabetes Study (UKPDS) and other major studies. Effects of SMBG on HbA(1c) came from clinical studies, meta-analyses and population studies, but can only be considered 'moderate' levels of evidence. Costs of complications were retrieved from published sources. Direct costs of diabetes complications and SMBG were projected over patient lifetimes from a UK National Health Service perspective. Outcomes were discounted at 3.5% annually. Extensive sensitivity analyses were performed. RESULTS: Depending on the type of diabetes treatment (diet and exercise/oral medications/insulin), improvements in glycaemic control with SMBG improved discounted QALYs anywhere from 0.165 to 0.255 years, with increased total costs of 1013 pounds sterlings- 2564 pounds sterlings/patient, giving incremental cost-effectiveness ratios of 4508 pounds sterlings: 15,515 pounds sterlings/QALY gained, well within current UK willingness-to-pay limits. Results were robust under a wide range of plausible assumptions. CONCLUSIONS: Based on the moderate level of clinical evidence available to date, improvements in glycaemic control with interventions, including SMBG, can improve patient outcomes, with acceptable cost-effectiveness ratios in the UK setting. PMID- 16709309 TI - Effect of multifactorial treatment on non-alcoholic fatty liver disease in metabolic syndrome: a randomised study. AB - BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is the hepatic manifestation of the metabolic syndrome (MetS). There is no established treatment for NAFLD. AIM: To evaluate a multifactorial intervention in the treatment of NAFLD. METHODS: A prospective, open-label, randomised study in non-diabetic patients (n = 186) with MetS (follow-up: 54 weeks). All patients had both biochemical and ultrasonographic evidence of NAFLD at baseline. Other causes of liver disease were excluded. Patients received lifestyle advice and treatment for hypertension (mainly inhibitors of the renin-angiotensin system), impaired fasting glucose (metformin), obesity (orlistat) and dyslipidaemia [randomly allocated to atorvastatin 20 mg/day (n = 63) or micronised fenofibrate 200 mg/day (n = 62) or both drugs (n = 61)]. Liver ultrasonography was assessed at baseline and at the end of the study. RESULTS: At the end of treatment, 67% of patients on atorvastatin, 42% on fenofibrate and 70% on combination treatment no longer had biochemical plus ultrasonographic evidence of NAFLD (p < 0.05 vs. baseline for all comparisons). The percentage of patients who no longer had evidence of NAFLD was significantly higher (p < 0.009) in the atorvastatin and combination groups compared with the fenofibrate group. This effect was independently related to drug treatment, as well as to reductions in high-sensitivity C-reactive protein, waist circumference, body weight, triglycerides, low-density lipoprotein cholesterol, total cholesterol, systolic blood pressure and glucose. Four patients discontinued treatment because of adverse effects. CONCLUSIONS: Multifactorial intervention in MetS patients with both biochemical and ultrasonographic evidence of NAFLD offsets surrogate markers of NAFLD (i.e. elevated aminotransferase plus echogenic liver). PMID- 16709311 TI - The cost-effectiveness of bimatoprost, latanoprost and timolol in treatment of primary open angle glaucoma in five European countries. AB - OBJECTIVE: The objective of this cost-effectiveness analysis is to evaluate cost effectiveness ratios of the intraocular pressure (IOP)-lowering agents bimatoprost, latanoprost and timolol in five major European countries: France, Germany, Italy, Spain and the UK. METHODS: The cost-effectiveness analysis is based on achievement of IOP targets between 13 and 18 mm Hg. Thus, the cost effectiveness ratios express the costs of having one patient successfully achieving IOP target. The perspective of the analysis is that of the health care sector payer, including costs of medicine and costs of ophthalmologist visits. The time frame is first year of glaucoma treatment. Four treatment strategies are analysed: Timolol as first line with add-on latanoprost or bimatoprost if IOP targets are not met, and latanoprost and bimatoprost as first line with add-on timolol. RESULTS: In the UK, Spain, Italy and Germany the timolol first with add on of bimatoprost is the least expensive treatment. This strategy dominates both strategies involving latanoprost (as add-on to timolol or as first line) in these four countries. The incremental cost-effectiveness ratio of bimatoprost first line therapy versus timolol with add-on bimatoprost varies from each country and target (from 305 pounds sterlings to 43,720 euros per patient). In France the timolol first line and latanoprost add-on is not dominated and is the cheapest alternative. The incremental cost-effectiveness ratio of timolol with add-on bimatoprost versus add-on latanoprost lies between 71 euros and 355 euros per patient depending on target (18 and 13 mm Hg, respectively). CONCLUSION: First line treatment of latanoprost is dominated in all countries. In four out of five countries the timolol first-line therapy with add-on latanoprost is also dominated. Based on this pharmacoeconomic analysis, the most cost-effective strategy seems to be timolol first line with add-on bimatoprost if target is not met after 3 months. PMID- 16709310 TI - Experience with starting tacrolimus postoperatively after transsternal extended thymectomy in patients with myasthenia gravis. AB - BACKGROUND: Thymectomy is a standard treatment of myasthenia gravis (MG). Immunomodulating agents are frequently given during the post-thymectomy latency period until complete remission is fully consolidated, but serious side effects is a relevant clinical problem for patients on long-term immunomodulating treatment. OBJECTIVE: To assess the effectiveness of starting tacrolimus in the immediate postoperative period in MG patients undergoing transsternal extended thymectomy, with complete stable remission (CSR) as the primary outcome of the study. METHODS: Forty-eight MG patients received tacrolimus, 0.1 mg/kg per day b.i.d. (started 24 h after thymectomy) and prednisone 1.5 mg/kg/day. Histologically, 34 patients had hyperplasia, 20 thymic involution, and 14 thymoma. Of the 48 patients, 40 completed 1 year of tacrolimus therapy, 38 completed 2 years, 27 completed 3 years, 21 completed 4 years, and 9 more than 5 years. Mean dose of tacrolimus was 4.9 mg/day (range 2-8 mg/day) with a mean plasma drug concentration of 7.6 ng/mL (range 7-9 ng/mL). Prednisone could be withdrawn after the first year in 93.7% of patients and at 2 years in 100%. RESULTS: The mean follow-up was 24.4 months, SD 17.3 (range 6-60 months). Improvement of muscular strength and decrease of anti-AChR antibodies were statistically significant (p < 0.001) shortly after operation. CSR was obtained in 33.4% of patients, pharmacological remission in 62.6%; 4% of patients had minimal symptoms. None of the patients with thymoma achieved CSR. The estimated median follow-up to obtain a CSR was 37.9 months (95% confidence interval [CI] 26.4-49.5 months). The overall crude CSR rate was 33.4%, with 47% for non-thymoma patients. The probability to achieve CSR at 3 years was 67% for the non thymomatous group. CONCLUSIONS: Long-term immune-directed treatment with tacrolimus to improve the effectiveness of thymectomy in MG is feasible and was associated with a high rate of CSR in patients without thymoma. PMID- 16709312 TI - Predictive factors and a corresponding treatment algorithm for controlled ovarian stimulation in patients treated with recombinant human follicle stimulating hormone (follitropin alfa) during assisted reproduction technology (ART) procedures. An analysis of 1378 patients. AB - BACKGROUND: Identifying parameters that can accurately predict the response to controlled ovarian stimulation (COS) would be of great benefit in assisted reproductive technology (ART) procedures. An analysis was undertaken with the objective of determining whether specific factors could optimally predict a response to stimulation in ART, and to then develop a corresponding treatment algorithm that could be used to calculate the optimal starting dose of recombinant human follicle stimulating hormone (r-hFSH; follitropin alfa) for selected patients. METHODS: The overall population consisted of 2280 normo ovulatory ART patients from 11 randomised clinical trials. However, for the final analysis population, only patients less than 35 years of age who received r-hFSH monotherapy (N = 1378) were included. RESULTS: Backwards stepwise regression modelling indicated that predictive factors for ovarian response included basal FSH, BMI, age and number of follicles < 11 mm at baseline screening. The concordance probability index was 59.5% for this model. CONCLUSIONS: In the largest data series so far analysed to determine predictive factors of ovarian response, basal FSH, BMI, age and number of follicles < 11 mm at screening were the most important variables in ART patients less than 35 years of age who were treated with r-hFSH monotherapy. Using these four predictive factors, a follitropin alfa starting dose calculator was developed that can be used to select the FSH starting dose required for an optimal response. The relevance of this dose calculator will be evaluated in a prospective clinical trial. PMID- 16709313 TI - Efficacy and safety of alendronate and risedronate for postmenopausal osteoporosis. AB - INTRODUCTION: This paper discusses the efficacy and safety of alendronate and risedronate in the treatment of postmenopausal osteoporosis. METHODS: The literature was searched with the PubMed from 1996 to the present, with respect to strictly conducted systematic reviews with homogeneity, meta-analyses with homogeneity, and randomized controlled trials (RCTs) with narrow Confidence Interval. RESULTS: According to the results of large randomized controlled trials (RCTs), bisphosphonates (alendronate, risedronate, and ibandronate), raloxifene, calcitonin, parathyroid hormone (PTH), and strontium ranelate effectively prevent vertebral fractures in postmenopausal women with osteoporosis. Because raloxifene has been shown to be effective in preventing the initial vertebral fracture in postmenopausal osteoporotic women without prevalent vertebral fractures, it is considered in the treatment of postmenopausal women with mild osteoporosis or osteopenia with some risk factors for fractures. RCTs have also demonstrated that alendronate, risedronate, PTH, and strontium are useful to prevent non-vertebral fractures and that alendronate and risedronate prevent hip fractures, thus alendronate or risedronate are primarily considered as the first-line drugs in the treatment of elderly women with osteoporosis having some risk factors for falls. While it has been suggested that PTH may be considered in patients with severe osteoporosis, the use of PTH in the treatment for osteoporosis is limited to 2 years or less, and it may be appropriate to use other anti-resorptive drugs after the completion of PTH treatment to maintain the skeletal effects gained during the treatment. RCTs have demonstrated that the incidence of gastrointestinal tract adverse events in postmenopausal osteoporotic women treated with bisphosphonates and placebo are similar, and also the long-term efficacy and safety of alendronate and risedronate. CONCLUSION: The evidence derived from the literature, based on strict evidence-based medicine guidelines, suggests that there is long-term efficacy and safety with alendronate and risedronate in the treatment of osteoporosis in postmenopausal women. PMID- 16709314 TI - A comparison of teriparatide and calcitonin therapy in postmenopausal Asian women with osteoporosis: a 6-month study. AB - OBJECTIVE: The number of hip fractures is expected to double in the next 20 years, with current estimates that Asia will account for 37% of these cases. As bone mineral density (BMD) may be used as a measure of fracture risk, we sought to compare the effects of teriparatide with salmon calcitonin treatment on changes in BMD, biochemical bone markers, and safety in postmenopausal Asian women with osteoporosis. METHODOLOGY: A total of 104 patients (n = 47 teriparatide [20 g/day subcutaneously] and n = 57 calcitonin [100 IU/day subcutaneously]) were enrolled in Hong Kong, Singapore, Philippines, Malaysia, and Thailand. Calcium (> or = 500 mg/day) and vitamin D (200-400 IU/day) supplements were taken throughout the 6-month controlled, randomized study. RESULTS: Teriparatide was associated with a 5.03 +/- 4.77% increase in lumbar spine BMD (p < 0.0001, mean +/- SD change from baseline), whereas changes in lumbar spine BMD for patients on calcitonin were not statistically significant (mean change of 0.36 +/- 4.12%, p = 0.16). Comparison of the two groups indicated that teriparatide treatment improved lumbar spine BMD statistically significantly more than calcitonin (p < 0.0001). No statistically significant changes were observed for total hip or femoral neck BMD. Serum bone-specific alkaline phosphatase (BSAP) increased by 55.9% (median change from baseline, p < 0.0001) in the teriparatide group, and remained stable with calcitonin (5.0% change, p = 0.24); osteocalcin increased by 156.15% (median change from baseline, p < 0.0001) with teriparatide, and decreased with calcitonin (-15.25%, p = 0.03). Similar rates of adverse events were observed, with nausea and dizziness the most commonly reported for both groups (teriparatide versus calcitonin, 13.0% versus 23.2% p = 0.21, 10.9% versus 21.4% p = 0.19, respectively). There were no clinically relevant changes observed in laboratory parameters. CONCLUSIONS: Both treatments were similarly tolerated, however teriparatide was associated with greater increases in lumbar spine BMD and bone formation markers, demonstrating the unique mechanism of action and safety of this treatment for osteoporosis in these Asian women. PMID- 16709315 TI - Correlation of improved erectile function and rate of successful intercourse with improved emotional well-being assessed with the Self-Esteem And Relationship questionnaire in men treated with sildenafil for erectile dysfunction and stratified by age. AB - BACKGROUND: The quality of life consequences of erectile dysfunction (ED) include depression, anxiety, and loss of self-esteem. The Self-Esteem And Relationship (SEAR) questionnaire is a validated, patient-administered, psychometric instrument specific to ED. OBJECTIVE: To determine correlations between erectile function (EF), intercourse success, and emotional well-being measured with the SEAR questionnaire in men treated with sildenafil citrate for ED and stratified by age (< 50 years, 50-65 years, and > 65 years). RESEARCH DESIGN AND METHODS: This was an open-label, flexible-dose trial of sildenafil (25, 50 and 100 mg) administered for 10 weeks to 382 men with ED (mean +/- SD age, 55 +/- 13 years; mean ED duration, 4 years), which was conducted at 62 centers in the United States. MAIN OUTCOME MEASURES: Analysis (by intent-to-treat, n = 368) of the change from baseline to the week-10 endpoint in the SEAR questionnaire Self Esteem subscale, the intercourse success rate (percent of occasions at which an erection that lasted long enough for successful intercourse was achieved), and their correlation. RESULTS: For the overall population, there was mean +/- SD improvement (p < 0.0001, paired t-tests) in the Self-Esteem subscale (56 +/- 25 to 79 +/- 22) and intercourse success rate (21 +/- 30% to 70 +/- 36%), which showed positive correlation (p < 0.0001). Secondary outcomes (i.e., EF domain of the International Index of Erectile Function; event log frequency of erection hard enough for sexual intercourse and of ejaculation/orgasm) also improved (p < 0.0001) and correlated positively with the SEAR Self-Esteem subscale and Sexual Relationship domain (p < 0.05 for all correlations). All 10 correlations were positive (p < 0.05) in men aged 50 to 65 years, eight were positive in men aged > 65 years, and six were positive in men aged < 50 years. The most common treatment related adverse events were mild-to-moderate headache (12% of patients), vasodilatation (7%), and rhinitis (4%). CONCLUSIONS: Men treated with sildenafil for ED demonstrated improved erectile function and an increased intercourse success rate, which correlated positively with improvement in SEAR measures of self-esteem and sexual relationship. PMID- 16709316 TI - An evaluation of patient preferences for osteoporosis medication attributes: results from the PREFER-US study. AB - OBJECTIVE: To evaluate preferences for eight medication attributes that women may consider when evaluating prescription osteoporosis medications. RESEARCH DESIGN AND METHODS: The eligible sample consisted of women aged 50 years or older who responded to the 2003 or 2004 Internet-based National Health and Wellness Survey as being diagnosed with osteoporosis, considering themselves at risk, or having a family history of osteoporosis. In this Internet survey (the PREFER survey), respondents were asked to: (1) force-rank order the eight attributes from one to eight according to their preferences and (2) separately rate the importance of each attribute on a Likert-type scale from 1 (extremely unimportant) to 7 (extremely important). RESULTS: We collected 999 responses across 3 days from a sample of 3368 women and stopped compiling responses after achieving sample size targets. Drug effectiveness (e.g., ability to reduce the risk of fractures) was force ranked as the No. 1 preferred osteoporosis medication attribute by 37% of the sample. Side effects were force ranked as the No. 1 preferred medication attribute by 36% of the sample. Dosing frequency, dosing procedure, and formulation (i.e., how the drug is taken) were each force ranked as No. 1 by 2% or less of the sample. Drug effectiveness had the highest percentage of 'extremely important' responses (59%) followed by drug interactions (53%). Drug effectiveness was the highest-rated attribute (mean [S.D.] = 6.1 [1.6], median = 7), while dosing frequency was the lowest-rated attribute (mean [S.D.] = 4.7 [1.8], median = 5). CONCLUSIONS: In our sample of women with a diagnosis of osteoporosis or at risk for osteoporosis, drug effectiveness was the most highly ranked and rated of eight osteoporosis medication attributes. Side effects and drug interactions were also highly ranked and rated. Healthcare providers should consider incorporating patient preferences for key medication attributes into their therapeutic decision-making processes. PMID- 16709317 TI - Open-label olanzapine treatment in bipolar I disorder: clinical and work functional outcomes. AB - OBJECTIVES: The objective of this study was to describe clinical and work functional outcomes associated with 6-month open-label olanzapine treatment for bipolar I disorder. METHODS: The study consisted of 249 patients entering a 6 month open label phase after 12 weeks of acute double-blind haloperidol or olanzapine treatment. Baseline for analysis was defined as the beginning of open label treatment. The clinical outcomes were symptomatic remission defined by a Y MRS total score < or = 12 and a HAM-D total score < or = 8 at the end of 6 months of treatment. The work functional outcomes included work functional scores, the proportion of patients who reported to 'work' as employee, volunteers, students, or house workers and the proportion of patients who specifically reported to 'work for pay'. RESULTS: A total of 240 patients reported work functional outcomes post open-label baseline. Among them, 15.4% patients moved into a 'work group' from a 'no-work group' at baseline, while 7.1% did the opposite (p = 0.0065) and 13.3% reported an improvement to 'work for pay' status from a 'not working for pay' status at baseline, while there was 4.2% of worsening in employment status (p = 0.0007). Overall, improvement in the work functional score was found at all post-baseline time points, beginning at month two (p = 0.003). LIMITATIONS: Results of this study need to be confirmed by double-blind randomized controlled studies. There was a lack of detailed information on work functioning from the questionnaire. CONCLUSIONS: Open-label olanzapine treatment for 6 months was associated with improvements in work functional outcomes in patients with bipolar disorder. PMID- 16709318 TI - Comments on Long-term clinical and cost outcomes of treatment with biphasic insulin aspart 30/70 versus insulin glargine in insulin-naive type 2 diabetes patients: cost-effectiveness analysis in the UK setting. PMID- 16709319 TI - Adjunctive glaucoma therapy use associated with travoprost, bimatoprost, and latanoprost. AB - PURPOSE: This study contrasts the utilization of adjunctive medication associated with travoprost, bimatoprost, or latanoprost, as primary glaucoma therapies. METHODS: Patients in the Medco Health database who initiated prostaglandin analog therapy on travoprost, bimatoprost or latanoprost between January 1, 2002 and July 31, 2002 were selected if they had no prostaglandin analog use in the prior 6 months. Patients were also required to have 12 months of prostaglandin therapy subsequent to the initial prescription. Data were available through July 31, 2003. The t-test and chi-square were used where appropriate to calculate p-values and assess significant differences. RESULTS: A total of 13 171 benefit-eligible subjects were identified of which 8381 (64%), 2637 (20%), and 2153 (16%) patients were treated with latanoprost, bimatoprost, and travoprost, respectively. There were no significant differences in mean age or gender between the three study groups with the exception that latanoprost patients were statistically older than travoprost patients (69.0 vs. 68.0). This was not considered a clinically meaningful difference. Overall, patients using travoprost or bimatoprost had a significantly lower rate of adjunctive medication use compared to patients starting on latanoprost monotherapy (22.5%, 23.2%, and 30.2 %, respectively). Therefore, for every 14 patients treated with latanoprost instead of travoprost or bimatoprost, one additional patient would be expected to need adjunctive therapy with another agent. The difference between travoprost and bimatoprost patients was not significant. CONCLUSIONS: The use of adjunctive medications to control intraocular pressure was significantly higher for latanoprost patients compared to travoprost and bimatoprost patients. This finding should be interpreted in the context that this study was based only on prescription claims data. It is important to simplify ophthalmic medical regimen as it is more cost effective, better for the patient, and minimizes the washout effect from administering two eye medications within 5 min. Decreasing the complexity of the patients' drug regimen may lead to increased adherence to prescribed therapy and a decreased risk of the incidence of blindness. PMID- 16709320 TI - Efficacy and safety of aceclofenac in the treatment of osteoarthritis: a randomized double-blind comparative clinical trial versus diclofenac - an Indian experience. AB - OBJECTIVE: Osteoarthritis is one of the most common forms of arthritis seen in primary care. Non-steroidal anti-inflammatory drugs (NSAIDs) play an important role in the management of osteoarthritis. However, gastrointestinal (GI) side effects limit their use. Cyclooxygenase-2 (COX-2) selective inhibitors exhibit better GI tolerability than conventional NSAIDs, but their cardiovascular safety is controversial. An NSAID with high efficacy, high GI tolerability and devoid of adverse cardiovascular effects is therefore a profile preferred by physicians. Aceclofenac is an anti-inflammatory and analgesic drug with preferential COX-2 inhibition. The objective of this study was to assess the efficacy and safety of aceclofenac in the treatment of osteoarthritis in an Indian population. RESEARCH DESIGN AND METHODS: The trial was controlled, comparative, randomized, and double blind. The study included 247 patients (82 males and 165 females, 40-82 years), suffering from osteoarthritis. Patients were randomized to receive either aceclofenac (100 mg twice daily) or diclofenac (75 mg twice daily). MAIN OUTCOME MEASURES: Clinical assessment was done at screening, randomization, and at 2 weeks, 4 weeks and 8 weeks of treatment by calculating Western Ontario MacMaster (WOMAC) scores, time taken to walk 100 feet, visual analogue scores for pain, investigator's assessment on a Likert scale and joint tenderness. Tolerability assessment was based on adverse events. Patient compliance was also assessed. RESULTS: Aceclofenac was found to be statistically superior to diclofenac in efficacy parameters of WOMAC scores, investigator's assessment and joint tenderness. Aceclofenac was found to be statistically superior to diclofenac in terms of epigastric discomfort, dyspepsia and abdominal pain. Compliance was also better with aceclofenac. The overall response of patients' osteoarthritis to aceclofenac was found to be statistically superior to diclofenac by both physician and patient. CONCLUSIONS: Aceclofenac is an effective and well tolerated drug in osteoarthritis in the Indian setting. PMID- 16709321 TI - Co-occurrence and comorbidities in patients with immune-mediated inflammatory disorders: an exploration using US healthcare claims data, 2001-2002. AB - OBJECTIVE: Research in immune-mediated inflammatory disorders (IMIDs) suggests that several diseases share disruptions in key cytokines. A common pathogenesis may present as similar patterns of disease co-occurrence and comorbidity, which could be observed through the analysis of healthcare claims datasets. METHODS: Adult patients continuously enrolled from 2001-2002 were identified in two US healthcare datasets containing medical and drug claims from health plans and self insured employers. Patients with treatment records indicating an IMID were selected (e.g., rheumatoid arthritis, psoriasis, Crohn's disease); controls for each disorder were matched 3:1 based on age, gender, region, and previous insurance coverage. IMID cohorts and comorbidities were identified using International Classification of Diseases, 9th revision codes. Prevalence relative risk was used to assess co-occurrence and comorbidity rates in IMID cohorts and controls. Medical and drug utilization patterns were also explored. RESULTS: Findings were similar across the two datasets. IMID patients represented about 4% of the population; specific IMID prevalence matched the epidemiology literature. Patients with at least one IMID had a higher risk for another IMID when compared to controls. The risk for infectious, renal, liver, and ulcerative comorbidities was also elevated. Selected drug utilization patterns confirmed comorbidity findings. IMID patients used more healthcare resources compared to controls; findings were robust under sensitivity analyses. CONCLUSIONS: IMID patients were generally more likely than controls to have another IMID, supporting the concept that the diseases are related. These patients also had higher comorbidity rates. Findings may be limited by the nature of claims datasets and the confounding effect of current treatments. Prospective studies are needed to determine whether IMIDs have a common pathogenesis. PMID- 16709322 TI - International INtegrated Database for the Evaluation of severe sePsis and drotrecogin alfa (activated) THerapy: component trials and statistical methods for INDEPTH. AB - OBJECTIVES: To better understand the effects of drotrecogin alfa (activated) (DrotAA) in severe sepsis patients, and the natural progression of severe sepsis, by creating a database of severe sepsis patients using the appropriate statistical analysis methods to integrate data from various trials. PATIENTS AND METHODS: Patient-level data from five severe sepsis trials, conducted by the same sponsor (Eli Lilly and Company, Indianapolis, IN, USA), were combined in an integrated database. Patients from various studies were included and received either DrotAA at 24 microg/kg/h for 96 hours (n = 3228) or placebo (n = 1231), in addition to standard supportive care. The following adjustments to the analyses were made to allow for the combined, and thus non-randomized, nature of the data: (1) differences in observed outcomes between studies were investigated to assess the extent of study-to-study variation before combining study-level data across trials for statistical analysis; (2) random study effects were included in models for patient-level data to capture potential extraneous study-to-study variation; and (3) propensity scores were computed and included as covariates in models for patient-level data to adjust for the nonrandomized nature of the data. RESULTS: Baseline characteristics were similar across the studies, supporting the combination of study-level data across trials. Comparing aggregate event rates between the two treatment arms yielded a relative risk for mortality (DrotAA versus placebo) of 0.79 (95% confidence interval [CI] 0.71-0.88), p < 0.0001. For patient-level analyses, after adjustment for 13 independent variables and random study effects, the odds ratio for mortality in the DrotAA versus placebo patients was 0.71 (95% CI 0.59-0.86), p = 0.0003. With adjustment for 13 independent variables and propensity score, the odds ratio was 0.79 (95% CI 0.67-0.93), p = 0.006. Limitations of this integrated database include the modest total number of the trials in the database and the fact that only one component trial in the database contributed data from both placebo and DrotAA-treated patients. SUMMARY: A robust severe sepsis database was developed which will be suitable for future studies on the progression of severe sepsis and the mechanism of action of DrotAA. Initial analysis of data from INDEPTH provides additional evidence that treatment of severe sepsis patients with DrotAA is associated with a sustained survival advantage throughout 28-day follow-up. PMID- 16709323 TI - Type 2 diabetes in youth from the Western Pacific region: glycaemic control, diabetes care and complications. AB - OBJECTIVE: To describe the glycaemic control, diabetes care and prevalence of complications in youth with type 2 diabetes from the Western Pacific Region. RESEARCH DESIGN AND METHODS: Cross-sectional, clinic-based audit of 331 patients aged < 18 years from 56 centres in Australia, China-Beijing, China-Shanghai, China-Hong Kong, Indonesia, Japan, South Korea, Malaysia, Philippines, Singapore, Taiwan and Thailand. Clinical and management data were recorded along with glycated haemoglobin (HbA(1c)), lipids and complication rates. MAIN OUTCOME MEASURES: Glycaemic control, complications, diabetes management. RESULTS: Median age was 14.9 years (interquartile range 13.2-16.4 years) and median diabetes duration 2.3 years (1.4-3.6 years). Median HbA(1c) was 7% (5.9-9.9%) and HbA(1c) was > 7.5% in 40% of patients. In multiple regression analysis, glycaemic control varied significantly between countries (p = 0.02); higher HbA(1c) was associated with fewer home blood glucose measurements (p = 0.005) and higher insulin dose/kg (p < 0.0001). Blood glucose monitoring was performed by 65% of patients (range 33 96% by country). In 25% of patients, management consisted of diet alone or no treatment (range 0-53% by country); oral anti-diabetic drugs alone were used in 49%, insulin alone in 11% and both in 15%. Microalbuminuria was found in 8% and hypertension in 24%. The risk of hypertension increased with higher BMI (OR 1.16, 95% CI 1.09-1.24, p < 0.0001); antihypertensive agents were used in 4% of patients. CONCLUSIONS: The management of type 2 diabetes in youth from the Western Pacific Region varies widely. Hypertension and microalbuminuria were frequent, but not commonly treated. Further investigation into the natural history and risk factors for complications in youth with type 2 diabetes is required to assist in developing evidence based management guidelines. PMID- 16709324 TI - Functional foods and paediatric gastro-intestinal health and disease. AB - The application of molecular methods to gastro-intestinal diseases is giving insight into the way in which the resident intestinal microbiota interacts with the mucosal immune system. Using traditional culture techniques, the importance of mucosally-associated bacterial biofilms in maintaining mucosal integrity has been demonstrated in ways previously impossible. Changes in the balance of organisms at initiation of and during disease provide a rationale for interventions with functional foods which facilitate re-establishment of the homeostasis of healthy gut. PMID- 16709325 TI - The role of Chlamydia pneumoniae in acute respiratory tract infections in young children in The Gambia, West Africa. AB - BACKGROUND: Little is known about the role of Chlamydia pneumoniae in the aetiology of acute respiratory tract infections (ARI) in children in developing countries. AIMS: To obtain better information, we studied the presence of C. pneumoniae and its association with clinical signs and symptoms of ARI in children under 5 years of age in The Gambia. METHODS: C. pneumoniae was sought by polymerase chain reaction in nasopharyngeal secretions and/or lung puncture aspirates from 324 infants under 3 months of age and 325 children between 3 months and 5 years of age with malnutrition, with or without pneumonia, and in control children. Clinical signs and symptoms for ARI and the spectrum of other viral and bacterial organisms were compared between those positive for C. pneumoniae and those negative. RESULTS: Of 324 young infants, ten (3.1%) showed the presence of C. pneumoniae whereas in the older children 50 of 325 (15%) were positive for C. pneumoniae. There was no significant association between clinical signs and symptoms of ARI and C. pneumoniae positivity in the young infants. Among older infants and children, there was a trend to more frequent lobar alveolar changes in those positive for C. pneumoniae. No bacterial pathogens were found to be significantly associated with C. pneumoniae infection. However, there was an association with measles in the malnutrition group and with RSV in the young infants group. CONCLUSIONS: In this study, C. pneumoniae was not associated with any particular clinical syndrome. We found no evidence that the organism plays a major role in ARI in young children in developing countries such as The Gambia. PMID- 16709327 TI - Tremors and thrombocytosis during treatment of megaloblastic anaemia. AB - In developing countries, a deficiency of cobalamine and folate contributes significantly to megaloblastic anaemia. Neurological observations in infants and young children with megaloblastic anaemia have included hypotonia, developmental regression, tremors and other abnormal movements. Following therapy with vitamin B12, coarse tremors occurred in six of 51 patients (12%) with megaloblastic anaemia. The tremors, which were noticed initially in the hands and feet, gradually became generalised and disappeared during sleep. They subsided within 5 11 days. Thirteen of 25 (52%) patients developed thrombocytosis between day 3 and week 5 of follow-up. In one child, the platelet count increased to >1300 x 10(9)/L. The importance of recognising these clinical findings during treatment of megaloblastic anaemia is emphasised. PMID- 16709326 TI - Reference values for oxygen saturation by pulse oximetry in healthy children at sea level in Chennai. AB - There is little information on oxygen saturation (SaO2) values in children in developing countries. AIM: To determine the reference values for oxygen saturation by pulse oximetry in healthy children living at sea level in Chennai and aged between 1 mth and 5 yrs. DESIGN AND SETTING: A prospective study was conducted in Kanchi Kamakoti CHILDS Trust Hospital from February to May 2005. METHODS: A total of 626 healthy children aged between 1 mth and 5 yrs were examined for heart rate, respiratory rate and SaO2. RESULTS: The mean SaO2 levels for children in the age groups 1-3 mths, 3 mths to 1 yr, 1-3 years and 3-5 years were 98.5%, 98.8%, 98.9% and 99.1%, respectively. The overall mean and median SaO(2) values for the children in the different age groups were 99%. The mean -2 SD values of oxygen saturation in the age groups were 96.5%, 96.4%, 96.3% and 97.1%, respectively, with an overall mean of 96.6%. CONCLUSION: The reference value for mean SaO2 in healthy children aged between 1 mth and 5 yrs and living at sea level in Chennai city was 98.5% or more, but the -2SD values had a mean of 96.6%. PMID- 16709328 TI - Polymorphisms in genes involved in folate metabolism as risk factors for oedematous severe childhood malnutrition: a hypothesis-generating study. AB - BACKGROUND: Severe childhood malnutrition (SCM) occurs as both oedematous and non oedematous syndromes. The reasons why some children develop oedematous SCM (OSCM) have remained elusive but differences in clinical presentation among malnourished children from similar backgrounds suggests that there might be inter-individual variation in susceptibility to OSCM. AIM: To estimate the strength of the association between variants of three genes involved in folate/methyl group metabolism [methylenetetrahydrofolate reductase (MTHFR), methionine synthase (MTR) and cystathionine beta-synthase (CBS)] and risk of OSCM. METHODS: Patients previously admitted to the Tropical Metabolism Research Unit (TMRU) for treatment of either OSCM (cases, n = 74) or non-oedematous SCM (NOSCM, controls, n = 50) were recruited. Genotypes at four sites within the three genes (MTHFR C677T, MTHFR A1298C, MTR A2756G and CBS 844ins68) were determined using PCR-based assays. RESULTS: The MTHFR 677T [odds ratio (OR) 0.63, 95% CI 0.2-1.7] and MTR 2756G (OR 0.74, 95% CI 0.4-1.4) alleles were associated with moderate reduction in risk of OSCM whereas the CBS 844ins68 allele (OR 1.4, 0.7-2.4) was associated with an increased risk. None of these risks was significant at the 5% level. CONCLUSIONS: Genetic variation within folate/methyl group metabolic pathways might have a small but potentially important influence on risk of OSCM. Additional, larger data-sets will be required to test the specific hypotheses (about the putative effect size and direction of association) generated in this preliminary study. Such observations have the potential to improve our understanding of the pathogenesis of clinical heterogeneity in severe malnutrition. PMID- 16709329 TI - Correlation of CD4 count, CD4% and HIV viral load with clinical manifestations of HIV in infected Indian children. AB - AIM: To correlate the absolute CD4 count, CD4% and HIV viral load with different clinical manifestations of HIV in antiretroviral-naive children. SETTING: The paediatric and perinatal HIV clinic in a tertiary care hospital over a period of 4 years, from January 1999 to December 2003. MATERIALS AND METHODS: A total of 92 highly active antiretroviral-naive, HIV-1-infected children were enrolled in a cross-sectional study. The clinical manifestations, age, sex and CDC classification of each patient were determined. CD4 count, CD4% and HIV-1 viral load were estimated at presentation and correlated with various clinical manifestations of HIV disease. RESULTS: CD4% was higher in infants (p < 0.001) and lower in children over 5 years of age (p = 0.01). Boys had a higher absolute CD4 count than girls (769 +/- 517 vs 532 +/- 430 cells/mm3, p = 0.02). Patients with lymphadenopathy (n = 43) had a high CD4 count (840 +/- 487 cells/mm3, p = 0.01) whereas patients with HIV cardiomyopathy (n = 4) had low CD4 counts (mean 182 cells/mm3, p = 0.04). In patients with failure to thrive (n = 29), the CD4% was low (14 +/- 9%, p = 0.02) and HIV-1 viral load was high (mean 4.5 x 10(5) copies/ml, p = 0.03). CD4 count, CD4% and HIV viral load did not correlate with the stage of the disease as per the CDC classification. CONCLUSION: HIV viral load, CD4 cell count and CD4% vary with age and disease complications in HIV infected children. However, CD4 count, CD4% and viral load did not correlate with CDC classification. PMID- 16709330 TI - Haematological abnormalities associated with paediatric HIV/AIDS in Lagos. AB - INTRODUCTION: In HIV-1-infected children, haematological disturbances include bone marrow abnormalities and peripheral cytopenias. All three major cell lineages can be depressed. METHODS: A cross-sectional study of baseline haematological parameters was undertaken in 68 children with confirmed HIV infection. In all cases, a complete blood count was done and some had CD4+ counts and HIV RNA PCR. The CD4+ count was analysed by the Coulter manual latex particle monoclonal antibody method and HIV RNA PCR by Roche Amplicor Monitor, version 1.5. RESULTS: Anaemia (< 100 g/L) was present in 77.9%, severe (< 60 g/L) in 5.9%, moderate (60-70 g/L) in 32.3% and mild (80-99 g/L) in 39.7%. The mean haemoglobin concentration decreased as disease progressed (p < 0.05); 6% had leucopenia, 17.5% had neutropenia and 2.5% (one case) had thrombocytopenia; also, the four (6%) subjects with leucopenia were in clinical stages B and C. Neutropenia, lymphocytopenia and thrombocytopenia were seen more in clinical stages B and C, though this relationship was not statistically significant. CONCLUSION: Both the erythroid and other cells lines are affected by HIV/AIDS and other associated factors. Anaemia is the most common haematological abnormality. The severity of peripheral cytopenias is related to the disease burden. PMID- 16709331 TI - Early-onset neonatal seizures: types, risk factors and short-term outcome. AB - In a large majority of term newborns, early-onset neonatal seizures (EONS) are believed to relate to perinatal risk factors. AIM: To identify risk factors for EONS. METHODS: Among a cohort of 1293 newborns admitted over a period of 2 years to the neonatal intensive care unit of Tikur Anbasa Hospital, Addis Ababa, 93 had seizures. The case control study method was used to identify risk factors associated with EONS. Univariate analysis was used to further examine risk factors after adjusting for the effect of severe perinatal asphyxia (Apgar < or = 3). RESULTS: A total of 78 (85%) term newborns had EONS. Hypoxic-ischaemic encephalopathy (OR 3.46, 95% CI 2.74-7.42) and shock (OR 2.53, 95% CI 1.51-4.76) were significantly associated with EONS. Multifocal clonic (66%) followed by focal clonic (22%) were the most common types of EONS. Nine (11%) of the newborns with EONS died. During follow-up, 37 (53%) of the 69 surviving newborns with EONS had psychomotor delay with or without neurological deficit. CONCLUSION: Hypoxic ischaemic encephalopathy and shock are important causes of EONS. PMID- 16709332 TI - Persistent umbilical discharge in infants and children. AB - BACKGROUND: Umbilical discharge is not an unusual presentation in infants and children. However, the evaluation and management are rarely discussed. METHOD: A retrospective review of 18 patients presenting with persistent umbilical discharge over a period of 22 years, excluding neonates with omphalitis. RESULTS: The patients' ages ranged between 11 days and 9 years. Presentation was usually discharge of clear, serous or purulent fluid or faecal matter from the umbilicus, sometimes dating back to the neonatal period. Evaluation consisted of bacteriology, fistulogram (one patient) and histopathology. The cause of discharge was patent vitello-intestinal duct in 11 patients, umbilical granuloma in three, umbilical sinus in two and patent urachus and osteomyelitis of the ischium in one each. Definitive treatment was exploration and complete excision of the lesion and antibiotics. Two patients with patent vitello-intestinal duct died, one before surgery from hypokalaemia and sepsis, and the other from anaesthetic complications. CONCLUSION: There is a need for early referral and treatment in patients with persistent umbilical discharge. Conservative treatment is usually not successful as the common causes would normally require complete excision. PMID- 16709333 TI - Lingual harmatoma in an infant. AB - Lingual tumours are uncommon in infants and harmatomas are rarely reported. A 5 month-old boy presented with a growth on the dorsum of the tongue which had been present from birth. The growth produced no respiratory or feeding difficulties. It was confirmed to be a harmatoma after complete excision. There has been no recurrence. PMID- 16709334 TI - Cerebral malaria owing to Plasmodium vivax: case report. AB - Cerebral malaria is a diffuse encephalopathy associated with seizures and status epilepticus which can occur in up to one-third of children with severe malaria, particularly that caused by Plasmodium falciparum. We report a unique case of P. vivax infection complicated by status epilepticus. A 4-year-old boy was admitted to the emergency outpatient clinic with intractable seizures for the previous hour. During resuscitation, two episodes of left-focal tonic convulsions occurred and he received phenytoin. Because of a history of P. vivax infection in the family 3 months previously, a blood slide for malaria was undertaken. Thin blood smears demonstrated P. vivax. He was treated with chloroquine and primaquine. An initial EEG was normal but after 3 months of follow-up a second EEG showed generalised epileptic activity. Rarely, cerebral malaria is a presenting complication or occurs during the course of P. vivax infection. In endemic areas such as Turkey, the possibility of cerebral malaria should be considered. PMID- 16709336 TI - Blueberry muffin syndrome owing to congenital rubella: case report. AB - A case of congenital rubella is reported in a 22-day-old boy presenting with a 'blueberry muffin' rash. Late-onset blueberry muffin syndrome following congenital rubella is very rare. PMID- 16709335 TI - Tubulo-interstitial nephritis and uveitis syndrome in a 6-year-old boy: case report. AB - A 6-year-old boy presented with a 4-month history of anorexia, weight loss and fatigue and was found to have tubulo-interstitial nephritis and uveitis (TINU) syndrome. After 2 months of corticosteroid therapy, resolution of the uveitis was almost complete but renal function showed only partial improvement. He was still in renal failure 6 months after diagnosis, but there was no relapse of the uveitis. Although the prognosis of TINU in children and adults is reported to be excellent, the outcome in young children can be less favourable, probably owing to delayed diagnosis and initiation of treatment. PMID- 16709337 TI - Recurrent epididymo-orchitis in an 8-year-old child with Kostmann syndrome (severe congenital neutropenia). AB - Underlying congenital genito-urinary tract anomalies are the most common cause of recurrent epididymo-orchitis in prepubertal boys. An 8-year-old boy was admitted with recurrent pulmonary and skin infections, was diagnosed as Kostmann syndrome and developed epididymo-orchitis. This appears to be the first case of Kostmann syndrome associated with epididymo-orchitis. PMID- 16709338 TI - Prevalence and clinical features of cystic fibrosis with pseudo-Bartter syndrome. PMID- 16709339 TI - Contact lens fitting in keratoconus. AB - The treatment of keratoconus can be implemented by the use of spectacle lenses, contact lenses of various kinds, and surgery. In this article, we deal with the procedures of contact lens fitting in patients diagnosed with keratoconus. The different types of specific contact lenses and the methodology of fitting these contact lenses for keratoconus are described. To select a contact lens for the individual patient depends upon the severity of keratoconus and the amount of corneal ectasia. Pending these criteria, a contact lens is chosen, which will give the best visual acuity and tolerance. At present, many designs of special contact lenses exist in oxygen-permeable materials; in addition, other systems, such as piggy-back, SoftPerm, semi-scleral, and scleral contact lens types are used. Contact lens fitting on a conical cornea will smooth out the highly irregular optical surface of the cornea and improve visual acuity considerably. Contact lenses are one of the better solutions to correct refractive errors induced by pathology, and they should be prescribed as soon as the keratoconus is detected to avoid development of amblyopia. The quality and quantity of vision is far better than with spectacle lens correction. PMID- 16709340 TI - "Special" intraocular lens designs: why do they sometimes fail? AB - A variety of new "special" intraocular lens models to optimize the visual performance after cataract surgery have been introduced. The majority of these new intraocular lens designs require a good centration within the capsular bag and/or capsular elasticity. Today, the main post-operative complications after cataract surgery still arise from retained lens epithelial cells within the capsular bag, resulting mainly in anterior and posterior capsule opacification. Some special intraocular lenses are not designed to be effective against proliferating lens epithelial cells. Although lens epithelial cells can never be completely removed from the capsular bag, surgical quality is a major factor for success or failure of these intraocular lens designs. PMID- 16709341 TI - Comments on: "special" intraocular lens designs: why do they sometimes fail? PMID- 16709343 TI - Comments on: screening for refractive errors in children. PMID- 16709342 TI - Screening for refractive errors in children. AB - It is well known that refractive anomalies play a prominent role in the genesis of amblyopia. The current aim of early visual screening is to look for these refractive anomalies along with strabismus and media opacities. This update reviews the frequency of refractive anomalies in the preschool-aged population, their amblyogenic potential, and the different methods of refractive screening, including the required equipment, costs, and diagnostic performance. PMID- 16709344 TI - Congenital nasolacrimal duct obstruction. AB - Congenital nasolacrimal duct obstruction is a common disorder in infants, with up to 20% of all newborns suffering from epiphora during the first year of life. This paper reviews the current evidence regarding the incidence, natural history, and management of congenital nasolacrimal duct obstruction. It highlights the role of nasendoscopy in improving the understanding of the condition, which has resulted in better overall management. PMID- 16709345 TI - Comments on: congenital nasolacrimal duct obstruction. PMID- 16709347 TI - Vision loss in one eye. PMID- 16709348 TI - The use of a "hybrid" trainer in an established laparoscopic skills program. AB - OBJECTIVES: Tabletop inanimate trainers have proven to be a safe, inexpensive, and convenient platform for developing laparoscopic skills. Historically, programs that utilize these trainers rely on subjective evaluation of errors and time as the only measures of performance. Virtual reality simulators offer more extensive data collection capability, but they are expensive and lack realism. This study reviews a new electronic proctor (EP), and its performance within the Rosser Top Gun Laparoscopic Skills and Suturing Program. This "hybrid" training device seeks to capture the strengths of both platforms by providing an affordable, reliable, realistic training arena with metrics to objectively evaluate performance. METHODS: An electronic proctor was designed for use in conjunction with drills from the Top Gun Program. The tabletop trainers used were outfitted with an automated electromechanically monitored task arena. Subjects performed 10 repetitions of each of 3 drills: "Cup Drop," "Triangle Transfer," and "Intracorporeal Suturing." In real time, this device evaluates for instrument targeting accuracy, economy of motion, and adherence to the rules of the exercises. A buzzer and flashing light serve to alert the student to inaccuracies and breaches of the defined skill transference parameters. RESULTS: Between July 2001 and June 2003, 117 subjects participated in courses. Seventy-three who met data evaluation criteria were assessed and compared with 744 surgeons who had previously taken the course. The total time to complete each task was significantly longer with the EP in place. The Cup Drop drill with the EP had a mean total time of 1661 seconds (average, 166.10) with 54.49 errors (average, 5.45) vs. 1252 seconds (average, 125.2) without the EP (P = 0.000, t = 6.735, df = 814). The Triangle Transfer drill mean total time was 556 seconds (average, 55.63) and 167.57 errors (average. 16.75) (EP) vs. 454 seconds (non-EP) (average. 45.4) (P = 0.000, t = 4.447, df = 814). The mean total times of the suturing task was 1777 seconds (average, 177.73) and 90.46 errors (average. 9.04) (EP) vs. 1682 seconds (non-EP) (average, 168.2) (P = 0.040, t = 1.150, df = 814). When compared with surgeons who had participated in the Top Gun course prior to EP, the participants in the study collectively scored in the 18.3th percentile with the Cup Drop drill, 22.6th percentile with the Triangle Transfer drill, and 36.7th percentile with the Intracorporeal Suturing exercise. When penalizing for errors recorded by the EP, participants scored collectively in the 9.9th, 0.1th, and 17.7th percentile, respectively. No equipment failures occurred, and the agenda of the course did not have to be modified to accommodate the new platform. CONCLUSIONS: The EP utilized during the Top Gun Course was introduced without modification of the core curriculum and experienced no device failures. This hybrid trainer offers a cost-effective inanimate simulator that brings quality performance monitoring to traditional inanimate trainers. It appears that the EP influenced student performance by alerting them to errors made, thus causing an increased awareness of and focus on precision and accuracy. This suggests that the EP could have internal guidance capabilities. However, validation studies must be done in the future. PMID- 16709349 TI - Laparoscopy training in United States obstetric and gynecology residency programs. AB - OBJECTIVES: To assess laparoscopic training curriculums in US Obstetrics and Gynecology residency programs. METHODS: A list of E-mail addresses was obtained for the accredited Obstetrics and Gynecology residency programs in the US from the CREOG Directory of Obstetric-Gynecologic Residency Programs and Directors. An E-mail survey containing 8 questions regarding laparoscopy training was sent to all residency directors with current E-mail addresses. RESULTS: Seventy-four residency directors responded to the survey for a response rate of 41%. Residency programs from all sections of the US were included in the study. Results of the survey indicate that 69% of residency programs had implemented a formal laparoscopy training program. At least half of the program directors surveyed stated that lack of faculty time and funds were the main barriers to laparoscopic surgery training. Seventy-two percent of those surveyed thought that in the future the health-care industry would demand proof of competency in laparoscopy as standard of care. CONCLUSIONS: Most US Obstetrics and Gynecology residency programs have implemented a formal laparoscopy training curriculum, use more than one method to train their residents, and involve almost half of their faculty on average in training residents to perform laparoscopic surgery. PMID- 16709350 TI - Core competency in laparoendoscopic surgery. AB - BACKGROUND: Concern about patient safety and physician competence was highlighted by the Institute of Medicine report, revealing the prevalence of fatal medical errors. There is also awareness that technical difficulties specific to laparoendoscopic surgery can cause medical errors. Reported herein is a review of the evidence pertaining to objective assessment of core competency components in laparoendoscopic surgery: cognitive skills, technical skills, surgical performance, and judgment. METHODS: PubMed and MedLine searches were performed to identify articles with combinations of the following key words: core competency, competency, laparoscopy, training, assessment, and curriculum. Further articles were obtained by searching reference lists of identified papers and through personal communication. CONCLUSIONS: The available evidence suggests that it is currently possible to objectively assess core competency components in laparoendoscopic surgery: knowledge and clinical judgment with well-established tests and innate technical abilities with computer-based simulators with embedded metrics. Simulation training is conducted to a proficiency criterion regardless of the number of repetitions or practice hours. Reports indicate that skills learned on a simulator transfer to the operating room. However, to date, objective assessment of surgical performance can be obtained only through review of unedited video tapes of surgical procedures by disinterested experts as recently demonstrated by our Japanese colleagues in urology. PMID- 16709351 TI - Video consent: a pilot study of informed consent in laparoscopic urology and its impact on patient satisfaction. AB - OBJECTIVES: In the current climate of increasing awareness, patients are demanding more knowledge of the operative process. We report a new protocol for consenting patients. In addition to the normal consent process, patients are invited to watch a video of the operation to gain a perspective of what is involved. We applied this novel method of consent and assessed its impact on patient satisfaction. METHODS: As part of postoperative follow-up, prospective data from 43 consecutive laparoscopic patients was obtained in the form of a self constructed, patient-directed questionnaire regarding the consenting process. Patients were also invited to complete the Client Satisfaction Questionnaire (CSQ 8) 4 weeks after their operation. Laparoscopic workload included nephrectomy (n = 27), deroofing of cysts (n = 2), pyeloplasty (n = 6), exploration for undescended testis (n = 6), lymph node dissection (n = 1), and nephropexy (n = 1). To reduce bias, an individual independent of the team treating the patient conducted these surveys. RESULTS: All study participants read the information leaflet before laparoscopic surgery, and 81% thought we should give patients the option to watch a video before their surgery. The leaflet information was understood by 92% of patients. All patients who opted to watch a video found it helpful to their understanding, and 75% requested a copy of the video of their own operation. The mean patient satisfaction (CSQ-8) score was 29.8 of a possible maximum score of 32. CONCLUSIONS: This novel approach to informed consent has had a positive impact on the patient journey as is evident from high satisfaction scores. Additional randomized, controlled trials need to be conducted to evaluate video consenting methods in laparoscopic urology. PMID- 16709352 TI - Classic intrafascial supracervical hysterectomy (CISH): 10-year experience. AB - OBJECTIVE: We report and review herein our 10-year experience with classic intrafascial supracervical hysterectomy focusing on our long-term experience, evolution of the operative technique, and increased use of this technique. METHOD: We performed a parallel, observational study with retrospective data to evaluate classic intrafascial supracervical hysterectomy, a laparoscopic hysterectomy technique, at Fayette Medical Center, a community hospital in Northwestern Alabama, USA. Patients comprised a consecutive series of 579 over a 10-year period from November 1992 through November 2002. RESULTS: The classic intrafascial supracervical hysterectomy technique, similar to standard supracervical hysterectomy, leaves the cardinal ligament, uterosacral ligament, vascular supply, and innervation to the upper vagina and cervix intact, but unlike supracervical hysterectomy removes the transition zone and endocervical canal. For 579 patients, the average age was 45.4 years (range, 22 to 92), follow up was 75.3 months (range, 17 to 137), operating room time was 69 minutes (range, 44 to 370), blood loss was 72 mL (range, 10 to 765), length of hospital stay was 23.2 hours (range, 14 hours to 5 days), time to return to work was 13.2 days (range, 3 to 28). Complications include 11 cervical bleedings, 1 uterine artery bleeding, 1 pelvic hematoma, 1 postoperative ileus, and 16 mucoceles of the cervical stump. Three patients were converted from a laparoscopic to an open procedure (0.52%). Long-term follow-up of up to 137 months shows no adverse events thus far. CONCLUSIONS: Classic intrafascial supracervical hysterectomy is a safe procedure with a low short- and long-term complication rate. It has a short recuperation period and high patient satisfaction. It is the procedure of choice when hysterectomy is indicated for benign disease. PMID- 16709353 TI - The treatment of mild endometriosis with laser laparoscopy: a two-step treatment analysis of patients whose primary therapy was successful. AB - OBJECTIVES: The most advantageous treatment for pelvic endometriosis that is not extensive has long been the subject of debate. In recent years, the ability to detect atypical presentations has allowed the gynecological surgeon to treat this entity more readily. The treatment in the past has been concerned with the singular treatment being applied at the time, not on the prior treatment effects on current therapy. The purpose of the current study was to see whether previous successful treatment modalities affected the success of subsequent laparoscopic laser fulguration treatment of endometriosis. METHODS: Patients who were previously treated for their endometriosis (minimal and mild) and had success in achieving pregnancy were treated for their disease after failing to become pregnant after delivery. They were treated with laparoscopic laser fulguration of the disease and the results were examined by chi-square (chi2) analysis. RESULTS: There appears to be no difference in current pregnancy rates in patients who were successfully treated in the original treatment for endometriosis, no matter which of the therapies had been used. CONCLUSION: If endometriosis is diagnosed at the time of laparoscopy and is easily amenable to treatment, it should be treated at the time of surgery regardless of prior treatment and results. PMID- 16709354 TI - Laparoscopic subtotal gastric resection for chronic gastric ulcers. AB - OBJECTIVES: We analyzed our experience with the laparoscopic approach for treating benign gastric lesions. METHODS: Between June 1998 and June 2002, we performed 18 gastric resections with the laparoscopic approach for 7 pyloric stenoses, 8 recurrent duodenal ulcers, and 3 chronic gastric ulcers. RESULTS: In our series, we performed Billroth II laparoscopic distal gastrectomy with no morbidity and mortality. CONCLUSIONS: Billroth II laparoscopic distal gastrectomy is safe in cases of benign gastric or duodenal lesions. PMID- 16709355 TI - Postoperative gastrointestinal complaints after laparoscopic Nissen fundoplication. AB - OBJECTIVES: Approximately 80% of patients complain of various symptoms immediately after laparoscopic Nissen fundoplication. These symptoms typically are treated medically without an extensive evaluation to identify the cause. We reviewed our experience of laparoscopic Nissen fundoplication to determine the course of postoperative symptomatology in our patient population, and present a rational approach to this problem. METHODS: Over a 10-year period, 628 patients underwent primary laparoscopic Nissen fundoplication for gastroesophageal reflux disease; patients were evaluated with a standard set of questions for postoperative gastrointestinal complaints. Three- and 6-month follow-up data were compared by using the chi square test. RESULTS: One-year follow-up data were available for 615 patients (98%). All of these patients had symptoms during the first 3 postoperative months. Early satiety (88%), bloating/flatulence (64%), and dysphagia (34%) were the most common; however, 94% of patients had resolution of their symptoms by the 1-year follow-up visit, and most had resolved after 3 months. Patients with persistent reflux or dysphagia after 3 months typically had an anatomic failure of the operation. CONCLUSIONS: Most patients who have undergone laparoscopic Nissen fundoplication for gastroesophageal reflux disease will have gastrointestinal complaints during the initial 3 postoperative months. Nearly all of these patients will have resolved their symptomatology after 3 months. Those with persistent symptoms after 3 months warrant evaluation for operative failure. PMID- 16709356 TI - Needlescopic cholecystectomy: lessons learned in 10 years of experience. AB - OBJECTIVES: Needlescopic cholecystectomy (NC) is a refinement of laparoscopic cholecystectomy (LC) using 2-mm instruments compared with the standard 5-mm and 10-mm ports. We review our experience with needlescopic cholecystectomy. METHODS: From 1994 to 2004, 303 patients underwent NC. All patients were operated on using 2-mm instruments and one 10-mm trocar for the laparoscope. The characteristics of patients, total operation time, complications, postoperative pain, and hospital course were documented. RESULTS: Patients' average age was 41.86 years; 262 were female and 41 were male. Mean BMI was 25.7. Mean length of surgery was 59.33 minutes. Intraoperative cholangiography was performed in all cases. Mean blood loss was 14.88 mL. One intraoperative complication occurred. Mean hospital stay was 22.68 hours. Postoperative pain was measured on a 0-10 pain scale; on day 0 it was 4.4 and on the first day it was 1.7. Analgesic doses required were 0 doses in 6.89%, 1 in 20.68%, 2 in 24.13%, 3 in 34.48%, 4 in 13.79%, and > 4 doses was not required. No postoperative complications occurred. At 3-month follow-up, patient satisfaction was 100%, and in 99% of patients scars were imperceptible. CONCLUSIONS: NC is safe and feasible without increased operative risk, with better cosmetic results, less pain, and good acceptance among patients. PMID- 16709357 TI - Laparoscopic cholecystectomy and management of biliary tract stones in a freestanding ambulatory surgery center. AB - OBJECTIVE: We conducted a retrospective 4-year study of patients undergoing laparoscopic cholecystectomy at a freestanding ambulatory surgery center. Data on rates of hospital admission, conversion to open surgery, bile duct injury, postoperative bile leakage, and incidence of choledocholithiasis were analyzed. The success rate for dynamic fluoroscopic intraoperative cholangiography was computed, and outpatient laparoscopic common bile duct exploration and anesthetic management were reviewed. METHODS: Patient charts from the ambulatory surgery center, office, and hospital were reviewed over a 4-year period commencing in October 1999. All cases were performed by 1 of 3 surgeons who are experienced with outpatient laparoscopic cholecystectomy and practice routine dynamic fluoroscopic intraoperative cholangiography. RESULTS: A total of 338 laparoscopic cholecystectomies were performed. Dynamic fluoroscopic intraoperative cholangiography was successfully performed in 89% (n = 302). No instances of bile duct injury or conversions to open surgery were reported. A 0.89% (n = 3) incidence of postoperative bile leak occurred. Six patients were admitted for inpatient care for a rate of 1.78%. Choledocholithiasis occurred in 2.0% and was managed successfully in the ambulatory setting. CONCLUSION: Laparoscopic cholecystectomy can be adapted to the freestanding ambulatory surgery environment with very high standards of care and very low complication rates. PMID- 16709358 TI - Minilaparoscopic appendectomy for acute appendicitis. AB - BACKGROUND: Minilaparoscopic appendectomy for appendicitis is not a well established procedure. This approach provides less abdominal wall trauma, fewer complications, and excellent cosmetic results. Our aim was to show the feasibility and safety of the minilaparoscopic approach. METHODS: Minilaparoscopic appendectomy was performed in 37 patients. Two 2.2-mm trocars were used to manipulate a 2.2-mm, 0-degree laparoscope and for grasper access. A 5-mm trocar was used for the ultrasonic scalpel. RESULTS: No deaths occurred. In 3 patients (8%), appendectomy was aborted due to pathology of the ovary. Conversion to the open approach occurred in 2.7% of patients. The average operating time was 34 minutes (range, 15 to 80), and the median length of hospital stay was 1.2 day (range, 1 to 5). CONCLUSIONS: The minilaparoscopic approach a) has the same advantages as the conventional laparoscopic approach in terms of better diagnostic accuracy and safety; b) a low incidence of complications; and c) yields excellent cosmetic results. PMID- 16709360 TI - Laparoscopic repair of umbilical hernias in conjunction with other laparoscopic procedures. AB - BACKGROUND: This study evaluates the feasibility of laparoscopic transfascial suture repair of umbilical hernias when combined with another laparoscopic procedure that potentially contaminates the peritoneal cavity. METHOD: From August 1997 to November 2001, 32 patients underwent laparoscopic umbilical suture repair in association with another laparoscopic procedure. The repair was performed with the Carter-Thomason suture passer. RESULTS: Of the 32, 26 patients with more than 1-year follow-up were included in the study. The mean diameter of the umbilical hernia defect was 1.67 cm (range, 0.5 to 3). At a mean follow-up of 34 months (range, 12 to 60), there were only 2 recurrences (7.7%) both of which happened in patients with hernia defects larger than 2 cm in diameter. Apart from 2 wound infections, no other complications occurred. CONCLUSION: Laparoscopic suture repair of umbilical hernias with the suture passer method is effective and durable even when combined with other laparoscopic procedures that potentially contaminate the peritoneal cavity with bile or enteric contents. PMID- 16709359 TI - Laparoscopic management of hepatic hydatid disease. AB - BACKGROUND: Hydatid disease is an endemic condition in several parts of the world. Owing to ease of travel, even surgeons in nonendemic areas encounter the disease and should be aware of its optimum treatment. A safe, new method of laparoscopic management of hepatic hydatid disease is described along with a review of the relevant literature. METHODS: Sixty-six cases of hepatic hydatid disease were operated on laparoscopically using the Palanivelu Hydatid System. The special trocar-cannula system used and the technique of operation are described. RESULTS: The majority of the patients presented with pain. Most of the patients had only a single cyst. The right lobe of the liver was most commonly involved. Cysts were bilateral in 4 patients. In 83.3%, simply evacuation of the hydatid cyst by the Palanivelu Hydatid System was done. In 13.7%, this was followed by a left lobectomy, as the cysts were large occupying almost the entire left lobe of the liver. The remnant cavity was dealt with by omentoplasty. The average follow-up period is 5.8 years. There have been no recurrences to date. CONCLUSION: We recommend Palanivelu Hydatid System for management of hepatic hydatid disease. We have found its efficacy to be optimum for preventing spillage, evacuating hydatid cyst contents, performing transcystic fenestration, and for dealing with cyst-biliary communications. PMID- 16709361 TI - Elective laparoscopic-assisted colectomy for sigmoid diverticulitis. AB - OBJECTIVES: The aim of this study was to evaluate the safety and effectiveness of laparoscopic-assisted sigmoid colectomy for diverticulitis and to assess its postoperative advantages. METHODS: From 1999 to 2001, 5 patients were selectively operated on with a laparoscopic-assisted procedure for uncomplicated sigmoid diverticulitis. In the preceding period (September 1997 through December 1998), 4 patients underwent open procedures for the same pathology. The surgical indication with the same criteria was restrictive: at least 2 acute episodes had occurred that were treated with hospital admission and that were separated by an adequate period (2 months) of medical therapy. RESULTS: No conversions of laparoscopy to an open procedure were necessary. Age, sex, weight, morbidity, and mortality were similar between the 2 groups. Operative time was 180 minutes for laparoscopy and 120 minutes for laparotomy. Postoperative resumption of peristalsis was 24 hours versus 4 days, resumption of alimentation was on the second postoperative day versus the fifth postoperative day, and hospital stay was 7 days versus 12 days for laparoscopy and laparotomy, respectively. CONCLUSION: This study shows the feasibility and the advantages of elective laparoscopic-assisted colonic resection for uncomplicated sigmoid diverticulitis. The advantages of the laparoscopic approach are the lower need for analgesics and the more precocious ambulation, canalization, resumption of alimentation, and the shorter hospital stay. PMID- 16709362 TI - Primary ovarian pregnancy after donor embryo transfer: early diagnosis and laparoscopic treatment. AB - Primary ovarian pregnancy is a rare type of ectopic pregnancy, particularly following in vitro fertilization. Although there have been a few reported cases of primary ovarian pregnancy following in vitro fertilization embryo transfer, we believe this is the first report involving donor embryo transfer. A high index of clinical suspicion, abnormal human chorionic gonadotropin levels, and early ultrasound evaluation may aid timely diagnosis and appropriate management. This report provides a reminder to practitioners to advise patients undergoing embryo transfer of the primary ovarian pregnancy risk. Information is provided herein regarding the diagnosis and management of primary ovarian pregnancy in women treated with in vitro fertilization. We review the criteria for early diagnosis and treatment options. PMID- 16709363 TI - Nodular adenomyosis of the uterus causing severe groin pain. AB - OBJECTIVES: To determine the cause of severe, disabling, groin pain. METHODS: We describe in this case report 2 patients with severe groin pain. Two myoma-like masses were found on the uterus near the right round ligament. Both masses were excised via laparoscopy. RESULTS: The pathology report of these 2 cases indicated adenomyosis. Both patients were asymptomatic after the procedure. Adenomyosis usually propagates in the myometrium, and the nodular shape of this pathology is rare. In patients with severe groin pain (menstrual or continuous), the gynecologist should think about both endometriosis of the round ligament and adenomyosis near the round ligament. CONCLUSION: Severe groin pain is rare but disabling. Endometriosis of the external part of the round ligament is described as a cause. In this case report, we explain that nodular adenomyosis can also be a cause. PMID- 16709364 TI - The incidence of retained fetal bone revealed in 2000 diagnostic hysteroscopies. AB - Three of 2000 diagnostic hysteroscopies revealed residual fetal bony fragments in women with abnormal uterine bleeding. Removal of bony fragments by hysteroscopy is associated with therapeutic success. PMID- 16709365 TI - Laparoscopic common bile duct exploration in pregnancy with acute gallstone pancreatitis. AB - BACKGROUND: We present a case in which a laparoscopic common bile duct exploration was performed safely in a pregnant patient with acute gallstone pancreatitis. CASE REPORT: A 25-year-old female, gravida 4 para 3, at 14-weeks gestation presented to her obstetrician with complaints of epigastric pain radiating to the back. She was otherwise healthy with no past medical or surgical history. A physical examination revealed a healthy young female with no evidence of jaundice and in no acute distress. An abdominal examination was remarkable for a gravid abdomen with mild tenderness to palpation in her epigastrium and negative Murphy's sign. The patient safely underwent a laparoscopic common bile duct exploration after a laparoscopic cholecystectomy was performed. CONCLUSION: This case illustrates the role of laparoscopic common bile duct exploration in the diagnosis and treatment of possible choledocholithiasis in a pregnant patient. PMID- 16709366 TI - Use of decellularized human skin to repair esophageal anastomotic leak in humans. AB - Multiple surgical techniques have been described for repair of esophageal leaks. None of the repairs are optimal, and the morbidity and mortality of an esophageal leak remains high. To our knowledge, this is the first case report of use of AlloDerm (Lifecell Corp, Brachburg, NJ) to successfully repair a thoracic anastomotic esophageal leak. PMID- 16709367 TI - Laparoscopic sleeve gastrectomy: an alternative for recurrent paraesophageal hernias in obese patients. AB - BACKGROUND: Recurrent paraesophageal hernias in obese patients are technically challenging and have a high recurrence rate. We sought to develop an alternative to the traditional approaches for this problem. This article describes the use of a sleeve gastrectomy in an obese patient with a large recurrent paraesophageal hernia. CASE REPORT: A morbidly obese 70-year-old woman presented with a 1-year history of chest pain, cough, dysphagia, and dyspnea. She had undergone an open paraesophageal hernia repair 8 years earlier. Diagnostic workup revealed a recurrent large paraesophageal hernia. Laparoscopically, we took down all adhesions, excised the hernia sac, reduced the stomach and distal esophagus into the abdomen, and closed the hiatus. We then resected the greater curvature and fundus of the stomach, leaving the lesser curve in a sleeve configuration. Eighteen months after the operation, the patient's chest pain, cough, dyspnea, and dysphagia were resolved. In addition, she has lost 57 pounds (255 to 198). CONCLUSION: A sleeve gastrectomy is a potentially useful alternative to fundoplication or gastropexy, or both of these, in the treatment of obese patients with complex paraesophageal hernias. PMID- 16709368 TI - Diagnostic laparoscopy and laparoscopic repair of a left paraduodenal hernia can shorten hospital stay. AB - Paraduodenal hernias are the most common forms of intraabdominal hernias, accounting for 53% of all internal hernias. However, these account for only 0.2% to 0.9% of all small intestinal obstructions overall. Patients usually report vague abdominal pains and discomfort lasting for many years. Furthermore, in patient diagnosis and management can last up to several weeks due to its rarity and unusual presentation. We report a case of a left paraduodenal hernia in an 18 year-old male who presented with abrupt onset of abdominal pain, nausea, and vomiting. He was subsequently managed by diagnostic laparoscopy and laparoscopic repair, which decreased the overall in-patient care to 2 days. PMID- 16709369 TI - Dumping syndrome as a complication of laparoscopic Nissen fundoplication in an adult. AB - BACKGROUND: Dumping syndrome is a recognized complication of various gastric surgical procedures, such as vagotomy, pyloroplasty, and gastrojejunostomy. However, it has not to date been reported following laparoscopic Nissen fundoplication in adults. CASE REPORT: We describe the case of a 34-year-old woman who developed late dumping syndrome following an uneventful laparoscopic Nissen fundoplication for gastroesophageal reflux disease. CONCLUSION: This condition should be considered in patients who develop hypoglycemic symptoms following laparoscopic fundoplication. PMID- 16709370 TI - Laparoscopic pancreaticoduodenectomy to treat cancer of the ampulla of Vater. AB - OBJECTIVES: We explored the feasibility, difficulty, and indications for laparoscopic pancreaticoduodenectomy. METHODS: Since November 11, 2002, we have successfully completed 5 laparoscopic pancreaticoduodenectomies. Patients included 4 males and 1 female, average age 43 years. Three patients had duodenal papillary cancer, one had cancer of the head of the pancreas, and one had pancreatic mixed cancer (duodenal papillary cancer, hepatobiliary ductal adenocarcinoma). The average mass size was 1.5/1.8 cm to 2.6/2.5 cm. RESULTS: The pathology diagnosis was well-differentiated duodenum papillary adenocarcinoma in 3 patients, head of pancreas endocrine small cell carcinoma in 1, and duodenum papillary adenoma with malignancy ductal intermediate differentiation adenocarcinoma in 1. During surgery, average blood loss was 770 mL. Operation time averaged 528 minutes. The main difficulties during surgery were estimation and identification of pancreatoduodenal tumor resection and hepatoduodenal ligament venation changes. After surgery, 1 patient had a small amount of pancreatic leakage, another developed stress ulcer bleeding; both patients became normal after appropriate treatment. The fourth patient developed severe recurrence of pancreatitis with pneumonia and on the 39th day after surgery developed stress ulcer bleeding. This patient died during the second operation. CONCLUSION: Laparoscopic pancreaticoduodenectomy is a very difficult and risky operation. It requires ample clinical experience in traditional pancreaticoduodenectomy, perfect laparoscopic surgery technique, consultation and cooperate with the surgical team, updated laparoscopy equipment, and very strict surgical indications. For hospitals that meet the above conditions and requirements, laparoscopic pancreaticoduodenectomy is very safe and feasible. PMID- 16709371 TI - Subphrenic and pleural abscess due to spilled gallstones. AB - BACKGROUND: A 70-year-old male approximately 3 years after laparoscopic cholecystectomy presented to his primary care physician with a 4-month history of generalized malaise. METHODS: A workup included magnetic resonance imaging that revealed a perihepatic abscess. The patient underwent ultrasound-guided drainage, with the removal of 1400 mL of purulent fluid and placement of 2 drains. Computed tomographic scanning showed resolution, and he was discharged home on oral antibiotics. At 2-month follow-up, the patient was asymptomatic, denying any constitutional symptoms. However, abdominal computed tomographic scanning revealed recurrence of the abscess, which measured approximately 18 x 9 x 7.5 cm, with mass effect on the liver. The patient was placed on intravenous antibiotics and scheduled for operative drainage. The abdomen was entered with a right subcostal incision, and 900 mL of purulent fluid was drained. We also noted abscess erosion through the inferolateral aspect of the right diaphragm into the pleural space. The pleural abscess was loculated and isolated from the lung parenchyma. Palpation within the abscess cavity revealed 9 large gallstones. Following copious irrigation and debridement of necrotic tissue, 3 drains were placed and the incision was closed. RESULTS: The patient had an uneventful recovery and was discharged home on postoperative day number 6. Follow-up imaging at 3 months demonstrated resolution of the collection. CONCLUSION: Spillage of gallstones is a complication of laparoscopic cholecystectomy, occurring in 6% to 16% of all cases. Retained stones rarely result in a problem, but when complications arise, aggressive surgical intervention is usually necessary. PMID- 16709372 TI - Abdominal wall abscess formation two years after laparoscopic cholecystectomy. AB - BACKGROUND: Spillage of gallstones within the subcutaneous tissue during laparoscopic cholecystecomy may lead to considerable morbidity. METHODS: We describe an abdominal wall abscess formation in a 50-year-old female that developed 24 months after a laparoscopic cholecystectomy. RESULTS: Spilled gallstones at the umbilical port site went undetected. Subsequently, an umbilical port-site abscess formed and was treated 2 years later. CONCLUSION: Any patient with a foreign body in the subcutaneous tissues after a laparoscopic cholecystectomy should be considered to have a retained stone. Use careful dissection, copious irrigation, and a retrieval device to avoid stone spillage. If spillage does occurs, percutaneous drainage and antibiotics followed by open retrieval of the stones should achieve adequate results during those delayed presentations of abdominal wall abscesses. PMID- 16709373 TI - Interval laparoscopic appendectomy for appendicitis complicated by pylephlebitis. AB - BACKGROUND: Although rare, portal mesenteric venous thrombosis and pylephlebitis remain potential life-threatening sequelae of ruptured appendicitis in children. Treatment recommendations from recent reports have included urgent exploratory laparotomy with appendectomy, prolonged intravenous antibiotic therapy, and anticoagulation for up to a year. METHODS: This report describes successful management of pylephlebitis and mesenteric venous thrombosis complicating ruptured appendicitis with intravenous antibiotics and anticoagulation followed by interval laparoscopic appendectomy. RESULTS: A previously healthy 5-year-old girl was diagnosed with ruptured appendicitis complicated by pylephlebitis and mesenteric venous thrombosis at the time of presentation. She was treated with intravenous antibiotics and anticoagulated for 3 months. She subsequently underwent interval laparoscopic appendectomy. At 3-year follow-up, she is healthy without evidence of adverse sequelae. DISCUSSION: This is the first reported case of successful, minimally invasive management of ruptured appendicitis complicated by mesenteric venous thrombosis and pylephlebitis. CONCLUSION: Similar treatment of other children with this rare presentation seems reasonable. PMID- 16709374 TI - Rare complications following colonoscopy: case reports of splenic rupture and appendicitis. AB - BACKGROUND: Appendicitis and splenic rupture are 2 rare complications of colonoscopy reported in the literature. To our knowledge splenic rupture following colonoscopy has been reported 17 times in the English-language literature and is associated with excess traction on the splenocolic ligament. Appendicitis after colonoscopy has been reported only 9 times and is usually associated with obstruction of the appendiceal lumen with fecal matter during colonoscopy. METHODS: We present the case reports of 2 patients: a 76-year-old woman who presented in consultation 24 hours after a routine colonoscopy with massive hemoperitoneum secondary to splenic rupture, seen on computed tomographic (CT) scan, who then underwent splenectomy; the second, a 60-year-old male who presented to the emergency room 16 hours after colonoscopy with clinical and computed tomographic scan findings of acute appendicitis who underwent a laparoscopic appendectomy. RESULTS: Treatment of both patients resulted in resolution of their complications, splenic rupture and appendicitis. They both had an uneventful postoperative course and are doing well several months postoperatively. CONCLUSIONS: We report 2 rare complications of colonoscopy, splenic rupture and appendicitis. In the setting of a recent colonoscopy and abdominal pain, a high index of suspicion is needed for their diagnosis. PMID- 16709375 TI - Laparoendoscopic removal of a benign gastric stromal tumor at the cardia. AB - OBJECTIVE: Gastrointestinal stromal tumors are the most common submucosal masses in the stomach and are mostly benign. Minimally invasive surgery is being increasingly used for their excision. Tumors close to the cardia often require a stapled resection of stomach. We report a technique for enucleating a 4-cm, well circumscribed gastric submucosal tumor at the cardia, avoiding gastric transection. METHODS: A gastroscope was introduced to distend the stomach. A laparoscope was inserted through the umbilicus after pneumoperitoneum was created. Two 5-mm metal trocars were inserted into the stomach under vision. A 10 mm trocar was passed through the umbilical incision into the stomach for the camera. Dilute epinephrine was injected submucosally. The tumor was enucleated after incising the overlying mucosa. A gastroscope snare helped in grasping the tumor for retraction and final removal in a plastic bag. The submucosal defect and gastric port-site defects were sutured laparoscopically with Vicryl 3-0. RESULTS: Contrast studies showed no leakage. Final histology indicated a benign leiomyoma. The patient was discharged on the sixth day. No recurrence was noted at 6-month follow-up. CONCLUSION: Benign stromal tumors at the cardia can be safely enucleated by this method. This technique is cost-effective as it avoids expensive staplers or self-retaining gastric balloon ports. PMID- 16709377 TI - Management of the retained percutaneous nephrostomy catheter. AB - Percutaneous nephrolithotomy is a widely accepted treatment for urinary calculi, but it is not without complications. We present the case of a 76-year-old male with a retained council tip catheter after percutaneous nephrolithotomy. Fluoroscopic guidance was used to perform percutaneous puncture of the catheter balloon, and the catheter was removed without complication. Advantages of various nephrostomy tube designs and additional measures to prevent this type of complication are discussed. PMID- 16709376 TI - Laparoscopic guidance or revision of ventriculoperitoneal shunts in children. AB - BACKGROUND: Ventriculoperitoneal shunt is the preferred treatment for hydrocephalus. Known complications include infection, obstruction, and disconnection with the fractured fragment migrating in the peritoneal cavity. We report 17 cases of laparoscopic evaluation and revision of ventriculoperitoneal shunts in children. METHODS: From January 2000 through October 2002, we retrospectively reviewed our experience with laparoscopy and ventriculoperitoneal shunts. RESULTS: Laparoscopy was performed in 17 children with a malfunctioning shunt, presumed shunt dislodgment or disconnection, reinsertion of a shunt after externalization, and primary shunt placement. Six patients (35%) were converted to an open laparotomy due to dense adhesions. Eleven patients (65%) underwent successful laparoscopic-assisted ventriculoperitoneal shunt placement: 5/11 (45%) had lysis of adhesions or pseudocyst marsupialization with repositioning of a functional shunt, or both; 3/11 (27%) had successful retrieval of a disconnected catheter with reinsertion of a new catheter; 2/11 (18%) had laparoscopic confirmation of satisfactory placement and function, requiring no revision; 1/11 (9%) had an initial shunt placed with laparoscopic guidance due to the obesity. Operative time for the laparoscopic procedure ranged from 30 minutes to 60 minutes. All laparoscopic procedures used 1-mm or two 5-mm ports. Perioperatively, no adverse neurological sequelae occurred due to the pneumoperitoneum. CONCLUSIONS: Laparoscopic guidance or revision of ventriculoperitoneal shunts permits (1) direct visualization of catheter insertion within the peritoneal cavity, (2) satisfactory positioning, (3) lysis of adhesions or marsupialization with catheter repositioning, or both, and (4) retrieval of fractured catheters. PMID- 16709378 TI - The Greenberg hysteroscopy speculum: a new instrument for hysteroscopy. AB - OBJECTIVES: Although the ubiquitous Graves speculum (open-sided or closed) is versatile and familiar, its shortest length is often too long for comfortable manipulation of the newer, pliable, flexible hysteroscopes. The Greenberg speculum is a modification of a Graves speculum with an open side to allow for the removal of instruments while they are still in the vagina. Whereas a standard mediumsized, open-sided Graves speculum measures 110 mm in maximum length, the Greenberg speculum measures only 74 mm. Although the difference between the speculums is only 36 mm in maximum length, this alteration can add considerable ease to placement of both flexible and rigid hysteroscopes in the office or operating room. In this study, we sought to quantify the benefit of this new device by demonstrating that this speculum brings the patient's cervix closer to the provider and reduces the length of unsupported hysteroscope shaft entering the cervical canal. METHODS: We measured the intraspeculum distance from the operator-side opening of the speculum to the external cervical os by using first a standard medium open-sided Graves speculum and then a Greenberg speculum. RESULTS: Use of the Greenberg speculum reduced the mean minimum intraspeculum distance from the extended fingertips grasping a hysteroscope to the external cervical os by an average of 28 mm or 34% compared with a Graves speculum. CONCLUSIONS: We believe this new speculum offers a subtle but distinct advantage over standard speculums for hysteroscopy. PMID- 16709379 TI - Gastric electrical stimulation: a novel treatment for gastroparesis. PMID- 16709380 TI - [From clinical trial to registry: the PRIAMHO II registry]. PMID- 16709381 TI - [Cardiac complications of non-cardiac surgery: initial approaches in the general low-risk population in Spain]. PMID- 16709382 TI - [Levosimendan in acute heart failure: past, present and future]. PMID- 16709383 TI - [Effect of combined beta-blocker and angiotensin-converting enzyme inhibitor treatment on 1-year survival after acute myocardial infarction: findings of the PRIAMHO-II registry]. AB - INTRODUCTION AND OBJECTIVES: Clinical trials have shown that combining beta blockers and angiotensin-converting enzyme (ACE) inhibitors has an additive effect in reducing mortality in patients with left ventricular dysfunction following acute myocardial infarction. Whether this additive effect also occurs in unselected post-myocardial infarction patients is unknown. METHODS: In total, 5397 patients who were discharged from hospital after suffering an acute myocardial infarction were followed for 1 year. The primary endpoint was all cause mortality. The effects of the medications on 1-year survival were analyzed using a Cox regression model, which included propensity scores for beta-blocker and ACE inhibitor use to take account of any potential imbalance in drug prescription rates. RESULTS: At hospital discharge, 55.9% of patients were receiving beta-blockers and 45.1%, ACE inhibitors. The 1-year mortality rate was 5.5%. Overall, combination of the two medications significantly reduced the 1 year mortality rate (hazard ratio [HR]=0.51; 95% confidence interval [IC], 0.32 0.82); P<.005) to a greater extent than ACE inhibitors alone (HR=0.78; 95% CI, 0.54-1.12; P=.2) or beta-blockers alone (HR=0.67; 95% CI, 0.43-1.05; P=.08). The same trend was also observed in low-risk patients without acute heart failure who had an ejection fraction > or =40%. CONCLUSIONS: In unselected post-myocardial infarction patients, combined prescription of beta-blockers and ACE inhibitors had an additive effect on the 1-year survival rate. PMID- 16709384 TI - [Invasive strategy in patients with advanced diabetes and non-ST-segment elevation acute coronary syndrome. Angiographic findings and clinical follow-up. PREDICAR study results]. AB - INTRODUCTION AND OBJECTIVES: Advanced diabetes can be associated with diffuse coronary artery disease that is difficult to treat by revascularization. We studied angiographic findings and disease progression in patients with advanced diabetes (either insulin-dependent or taking antidiabetic drugs for >5 years) and non-ST-elevation acute coronary syndrome who were being treated using an invasive strategy. METHODS: The study included 141 patients. The extent of the coronary artery disease was quantified using a score derived from a 29-segment coronary angiogram. The composite endpoint was death, myocardial infarction, or readmission for unstable angina within one year of follow-up. RESULTS: The extent of coronary disease was associated with Killip class >1 at admission (P=.02), previous coronary surgery (P=.003), ST-segment depression (P=.01), and a poor ejection fraction (P=.0001). The more of these factors present (i.e., 0, 1, 2 or 3 factors), the greater the extent of the coronary disease (i.e., 12 [7], 15 [7], 21 [6] and 23 [7] points, respectively; P=.0001). There was a significant difference between patients with > or =2 factors and those with P=.02), even after adjustment using a revascularization propensity score (C-index 0.80). CONCLUSIONS: In patients with non-ST-elevation acute coronary syndrome and advanced diabetes being managed using an invasive strategy, a history of coronary surgery, ST-segment depression and poor left ventricular function were all associated with the presence of diffuse coronary artery disease. Clinical follow up indicated that revascularization during hospital admission improved prognosis. PMID- 16709385 TI - [Cardiac complications of major elective non-cardiac surgery: incidence and risk factors]. AB - INTRODUCTION AND OBJECTIVES: Previous estimates of the incidence of cardiac complications following major elective surgery have all been obtained in populations with specific risk factors or in contexts that differ from those found in Spain. Our aims were to estimate the frequency of postoperative cardiac complications following major elective non-cardiac surgery in patients aged 40 years or more, and to identify associated risk factors. METHODS: Prospective observational study of consecutive patients undergoing surgery between November 1997 and October 1999. RESULTS: The overall incidence of postoperative cardiac complications was 9.6 (95% CI, 5.3-16): it was 1.9 (95% CI, 0.5-7.5) for superficial surgery, 36.2 (95% CI, 15.1-87) for deep surgery, and 150 (95% CI, 67.4-334) for peripheral vascular surgery. The risk of cardiac complications was greater in patients aged 75 years or more (relative risk [RR]=9.5; 95% CI, 2.6 34.9), in those with type-I diabetes (RR=7.1; 95% CI, 2.1-24.1), systolic blood pressure > or =180 mmHg (RR=5.8; 95% CI, 1.3-25.4), or electrocardiographic signs of ischemia (RR=25.3; 95% CI, 6-106.8), in those who had undergone deep or peripheral vascular surgery (RR=21; 95% CI, 5.4-81.6), and in those who experienced hypotensive episodes during surgery (RR=8.9; 95% CI, 2-39.1). CONCLUSIONS: Cardiac complications occurred in 1% of consecutive patients aged 40 years or more in the three months following major elective non-cardiac surgery. PMID- 16709386 TI - [Hemodynamic effects of levosimendan compared with dobutamine in patients with low cardiac output after cardiac surgery]. AB - INTRODUCTION AND OBJECTIVES: Levosimendan is an inotropic agent that is effective in the treatment of heart failure. However, experience with levosimendan in patients with reduced cardiac output following cardiopulmonary bypass is limited. The objective of this study was to compare the short-term hemodynamic effects of levosimendan with those of dobutamine in managing low cardiac output after cardiac surgery. METHODS: Forty-one patients who had low cardiac output after cardiopulmonary bypass were randomly assigned to dobutamine (n=20), 24-hour infusion of 7.5 microg/kg per min, or levosimendan (n=21), at a loading dose of 12 microg/kg followed by 24-hour infusion of 0.2 microg/kg per min. The following parameters were determined during a 48-hour observation period: arterial, central venous, pulmonary arterial and pulmonary capillary wedge pressure, cardiac index, heart rate, stroke volume, and systemic and pulmonary vascular resistance. RESULTS: Although both dobutamine and levosimendan improved the cardiac index, the increase was significantly greater with levosimendan (2.4 [0.2] l/min per m2 vs 2.9 [0.3] l/min per m2, respectively, at 24 h; P<.05). Moreover, levosimendan significantly reduced systemic and pulmonary vascular resistance, and significantly decreased systemic arterial, pulmonary arterial, pulmonary capillary wedge, and central venous pressure. CONCLUSIONS: Both dobutamine and levosimendan are effective in managing postoperative low cardiac output. However, levosimendan induces non-specific systemic, venous and pulmonary vasodilation which can result in hypotension as a adverse event. In these patients, it is advisable to omit or reduce the loading dose. PMID- 16709387 TI - [Heart failure with preserved ejection fraction. Effect of etiology on prognosis]. AB - INTRODUCTION AND OBJECTIVES: Heart failure with preserved systolic function accounts for almost 40% of heart failure cases. Prognosis is similar to that in patients with a low left ventricular ejection fraction (LVEF). However, it is not clear whether the etiology of heart failure with preserved systolic function has an effect on prognosis. METHODS: We assessed 95 consecutive patients admitted to our hospital with heart failure and a LVEF>45%. Twenty-five (26%) had an ischemic etiology and 70 (74%), a non-ischemic etiology. RESULT: The patients' mean age was 73 (6) years, 60% were female, and their mean LVEF was 61 (7)%. These characteristics were similar in the two etiological groups. After a mean follow up period of 53 (8) months (4-69 months; median 46 months), mortality was higher in ischemic patients (17.88 vs 2.37/100 patient-years; P<.0001), as was the rate of cardiovascular admissions (24.58 vs 4.14/100 patient-years; P<.0001). The rates of mortality due to heart failure and sudden death were also higher in ischemic patients, at 7.82 vs 0.59/100 patient-years, and 7.82 vs 0.30/100 patient-years, respectively (P<.0001). The higher overall admission rate found in the ischemic group was due to higher rates of admission for heart failure (14.53 vs 0.89/100 patient-years; P<.0001) and acute coronary syndrome (8.94 vs 1.78/100 patient-years; P=.003). CONCLUSIONS: In terms of prognosis, heart failure with preserved systolic function is not a homogeneous disease entity. Morbidity and mortality rates are higher in patients with an ischemic etiology. Moreover, different mechanisms are involved. PMID- 16709389 TI - [Congenital coronary artery anomalies with origin in the contralateral sinus of Valsalva: which approach should we take?]. AB - Recent years have witnessed a change in our perception of congenital coronary artery anomalies. From being regarded as simple coronary angiographic observations of little clinical significance, they have come to be seen as potential causes of sudden death in young people and of myocardial ischemia in adults. Diagnostic difficulties, a lack of knowledge about the mechanisms through which they produce myocardial ischemia, and their highly variable incidence have made congenital coronary artery anomalies of great interest to cardiologists. This article focuses on the group of coronary artery anomalies that is responsible for most clinical events: anomalies that have their origin in the contralateral sinus of Valsalva. Coronary artery anomalies are defined, their incidence is discussed, diagnostic criteria are given, and finally a treatment algorithm based on current knowledge is proposed. PMID- 16709388 TI - [Value of coronary blood flow pattern as a predictor of functional recovery and short-term left ventricular remodeling after primary coronary angioplasty. A transthoracic Doppler study]. AB - INTRODUCTION AND OBJECTIVES: Coronary blood flow measurement using a Doppler guidewire is the most sensitive way of detecting the no-reflow phenomenon following reperfusion of a myocardial infarction (MI). New high-frequency Doppler probes enable coronary blood flow velocity to be measured noninvasively. Our aims were to study the different patterns of left anterior coronary artery blood flow observed by transthoracic Doppler echocardiography, and to describe their association with functional recovery following reperfusion of an anterior MI. METHODS: The study included 57 patients with a mean age of 60 years (range 30-85 years). An abnormal coronary blo:d flow pattern was defined as one in which there was a high peak diastolic velocity and a short deceleration time (i.e., < or = 500 ms). We compared the regional contractility, ventricular volumes, and left ventricular ejection fraction (LVEF) measured after 72 hours with those measured 1 month after MI. RESULTS: Overall, 31 patients (54%) had a normal coronary blood flow pattern (Group 1) and 26 (46%), an abnormal pattern (Group 2). After one month, regional contractility improved in Group-1 patients, as did LVEF, from 46.8 (8.6) to 52.6 (8.8)% (P=.002). In these patients, left ventricular volumes were unchanged. In contrast, regional contractility and LVEF remained unchanged in Group-2 patients whereas ventricular volumes increased, from 55.8 (12.9) to 62.9 (16.8) ml/m2 (P=.05), and from 32.2 (9.5) to 37.1 (14.9) ml/m2 (P< .05). Coronary blood flow pattern was the most important independent predictor of left ventricular remodeling, odds ratio =6.14 (95% CI, 1.56-24.17). CONCLUSIONS: Transthoracic Doppler echocardiographic assessment of coronary blood flow following reperfusion of an anterior myocardial infarction can be used to identify patients with microvascular damage who are progressing towards ventricular dilatation without recovery of myocardial function. PMID- 16709390 TI - [Ischemic heart disease in women: clinical presentation, non-invasive testing and management of acute coronary syndromes]. AB - Cardiovascular disease is the leading cause of death in women. Because of the protective effects of estrogens, ischemic heart disease occurs predominantly in the postmenopausal period and, therefore, much later than in men. The proportion of women with acute coronary syndromes who present with typical chest pain is the same as in men. Nevertheless, atypical chest pain, which is usually associated with a lower likelihood of significant coronary disease, occurs more frequently in women. It is within this context that the sensitivity and specificity of the non-invasive tests used for diagnosing ischemic heart disease are reviewed. In addition, the clinical characteristics, treatment and prognosis of acute coronary syndromes in women are described, while highlighting the contributions made by age and disease severity at presentation to the higher short-term mortality observed in female patients after acute myocardial infarction. PMID- 16709391 TI - [Images in cardiology. CT angiography with three-dimensional reconstruction of vascular rings]. PMID- 16709392 TI - [Influence of diabetes mellitus on the management and prognosis of non-ST elevation acute coronary syndrome]. AB - The presence of diabetes mellitus worsens prognosis in acute coronary syndromes. The aim of our study was to analyze retrospectively the influence of diabetes mellitus on the management and prognosis of patients with non-ST-segment elevation acute coronary syndrome. We compared the baseline clinical characteristics of 273 patients (93 diabetic and 180 non-diabetic) admitted consecutively to our department with the diagnosis of non-ST-segment elevation acute coronary syndrome. In both groups, we assessed the medical treatment given during hospitalization and the use of coronary angiography, percutaneous coronary intervention, and coronary artery bypass grafting. Finally, we determined the incidence of heart failure during hospitalization and mortality at 28 days and 6 months in both groups. Multifactorial analysis revealed that diabetes was an independent risk factor for mortality during the study period. Data from our registry indicate that these findings were not associated with more extensive use of interventions in diabetic patients. PMID- 16709393 TI - [Kawasaki disease: heart disease during childhood]. AB - The aim of this study was to determine risk factors for coronary lesions, the type of heart lesion present, and long-term outcome in Kawasaki disease. We studied 150 children, aged 3 months to 9.5 years, who met the criteria for a diagnosis of Kawasaki disease. Of the 18% who were diagnosed with heart disease, 100% had coronary artery abnormalities, 11.1% had pancarditis, and 3.7%, mitral insufficiency. The coronary abnormalities were classified according severity as follows: diffuse ectasia in 40.7%; a small-to-medium-sized solitary coronary artery aneurysm in 33.3%; numerous small-to-medium-sized aneurysms in 11.1%; giant aneurysms in 11.1%; and coronary artery stenosis in 3.7%. The presence of pancarditis was a predictor of a giant coronary artery aneurysm. Mortality was 3.7%. Coronary aneurysm was the predominant heart lesion. The risk factors for coronary aneurysm in Kawasaki disease included age less than 27 months, fever lasting more than 8 days, erythrocyte sedimentation greater than 70 mm, and pancarditis. PMID- 16709394 TI - [Catheterization and vascular infection]. AB - The infectious complications that occur following vascular access for catheterization are frequently associated with an unfavourable outcome, mainly because diagnosis is delayed. Although its incidence is low, clinicians should be aware of this complication as the use of invasive procedures is increasing. Here, we report our experience with five patients. PMID- 16709395 TI - [Infective endarteritis in patent ductus arteriosus and septic pulmonary embolism]. PMID- 16709396 TI - [Hypertensive heart disease: a proposed clinical classification]. PMID- 16709397 TI - [Contrast echocardiography and clinical safety]. PMID- 16709398 TI - [Underdiagnosed cardiac cephalgia?]. PMID- 16709400 TI - Prestin is expressed on the whole outer hair cell basolateral surface. AB - Prestin has been identified as a motor protein responsible for outer hair cell (OHC) electromotility. Previous experiments revealed that OHC electromotility and its associated nonlinear capacitance resided in the OHC lateral wall and was not detected at the apical cuticular plate and basal region. In this experiment, the distribution of prestin in adult mouse, rat, and guinea pig OHCs was re-examined by use of immunofluorescent staining and confocal microscopy. We found that prestin labeling was located at the whole OHC basolateral wall, including the basal plasma membrane. However, staining at the basal membrane was weak. As compared with the intensity at the lateral wall, the intensities of prestin labeling at the membrane at the nuclear level and basal pole were 80.5% and 61.1%, respectively. Prestin labeling was not found at the cuticular plate and stereocilia. The prestin labeling was also absent in the cytoplasm and nuclei. The OHC lateral wall above the nuclear level is composed of the plasma membrane, cortical lattice, and subsurface cisternae. By co-staining with di-8-ANEPPS, prestin labeling was found at the outer layer of the OHC lateral wall, which was further evidenced by use of a hypotonic challenge to separate the plasma membrane from the underlying subsurface cisternae. The data revealed that prestin is expressed at the whole OHC basolateral membrane. Prestin in the basal plasma membrane may provide a reservoir on the OHC surface for prestin-recycling and may also facilitate performing its hypothesized transporter function. PMID- 16709401 TI - Verbal and spatial working memory in older individuals: A positron emission tomography study. AB - Recent reviews of a substantial number of studies have partially resolved questions concerning the brain regions used by working memory for manipulation and representation. We report a large single experiment in middle-aged to older adults (n = 89), classified by hypertensive status. Our design addresses the question of regions related to manipulation and representation, most particularly comparing spatial and verbal working memory. A control, memory search, and 2-back running memory task were performed with identical stimuli and responses during whole-brain 15O water positron emission tomography (PET) scans. Letter or spatial position instructions created verbal or spatial working memory versions of the tasks. We assessed agreement with the literature using regions of interest that were defined by clusters of activation empirically derived from the literature by Wager and Smith (Wager, T.D. and Smith, E.E., Neuroimaging studies of working memory: a meta-analysis, Cognitive, Affective and Behavioral Neuroscience, 3 (2003) 255-274). Our results largely confirmed conclusions from the review on the organization of working memory into dorsal prefrontal manipulation and ventrolateral prefrontal maintenance areas and representation in dorsal and ventral paths. Specific verbal versus spatial comparisons were also concordant with prior work establishing posterior lateralized representation for different contents by working memory. The similarity of results between this older sample and results derived by others from younger participants is notable. PMID- 16709402 TI - Neuroprotection by pyrroloquinoline quinone (PQQ) in reversible middle cerebral artery occlusion in the adult rat. AB - Pyrroloquinoline quinone (PQQ) is a naturally occurring redox cofactor that acts as an essential nutrient, antioxidant, and redox modulator. It has previously been reported to reduce infarct size in 7-day-old rat pups with an in vivo cerebral hypoxia/ischemia model (Jensen et al., 1994). In this study, we tested whether improvement is found in both behavioral measures of protection and by histological measures of infarcted tissue at 72 h after reversible middle cerebral artery occlusion (rMCAo) in adult rats. Two-hour rMCAo was induced in adult rats using the intraluminal suture technique. PQQ (10, 3, and 1 mg/kg) was given once by intravenous injection at the initiation, or 3 h after the initiation, of 2 h rMCAo. Neurobehavioral deficits were evaluated daily for 3 days followed by infarct volumes measurements by 2,3,5-triphenyltetrazolium chloride (TTC) staining. PQQ at 10 mg/kg infused at the initiation, or 3 h after the initiation, of rMCAo was effective in reducing cerebral infarct volumes measured 72 h later. At 3 h after ischemia, a dose of 3 mg/kg significantly reduced infarct volume compared to vehicle-treated animals, but 1 mg/kg was ineffective. Neurobehavioral scores were also significantly better in the PQQ treated group compared to the vehicle controls when PQQ was given at 10 and 3 mg/kg, but not at 1 mg/kg. Thus, PQQ is neuroprotective when given as a single administration at least 3 h after initiation of rMCAo. These data indicate that PQQ may be a useful neuroprotectant in stroke therapy. PMID- 16709403 TI - NR3 protein expression in trigeminal neurons during postnatal development. AB - The N-methyl-d-aspartate (NMDA) receptor plays an important role in the generation of rhythmical oral motor activities. To compliment our previous studies, we examined the developmental regulation of NR3A and NR3B expression in trigeminal motoneurons (Mo5) and mesencephalic trigeminal neurons (Me5). NR3A immunoreactive neurons were observed at all ages in both nuclei, decreasing in Mo5 and caudal Me5 after P14, and increasing in rostral Me5. NR3B protein expression only emerged in Mo5 after P21-23. Results indicate that NR3A and NR3B expression is differentially regulated between Mo5 and Me5 coincident with the transition from suckling to chewing. PMID- 16709404 TI - Synthesis, nucleic acid hybridization properties and molecular modelling studies of conformationally restricted 3'-O,4'-C-methylene-linked alpha-L ribonucleotides. AB - Nucleotides with conformationally restricted carbohydrate rings such as locked nucleic acid (LNA), alpha-L-LNA or 2',5'-linked 3'-O,4'-C-methyleribonucleotides exhibit significant potential as building blocks for antigene and antisense strategies. 2',5'-Linked alpha-L-ribo configured monomer X (termed alpha-L-ONA) was designed as a potential structural mimic of alpha-L-LNA. The corresponding phosphoramidite building block of monomer X was obtained in five steps (10% overall yield) from the easily obtainable thymine derivative 1. Incorporation of monomer X into oligodeoxyribonucleotides (ONs) results in dramatically decreased thermal stabilities with DNA/RNA complements (DeltaTm/mod=-11.5 to -17.0 degrees C) compared to unmodified reference ONs. Less pronounced decreases (DeltaTm/mod= 4.5 to -8.5 degrees C) are observed when monomer X is incorporated into triplex forming ONs and targeted against double-stranded DNA (parallel orientation, pyrimidine motif). This biophysical data, together with modelling studies, suggest that 2',5'-linked alpha-L-ONA is a poor structural mimic of alpha-L-LNA. PMID- 16709406 TI - On the emergence of modern humans. AB - The emergence of modern humans with their extraordinary cognitive capacities is ascribed to a novel type of cognitive computational process (sustained non routine multi-level operations) required for abstract projectuality, held to be the common denominator of the cognitive capacities specific to modern humans. A brain operation (latching) that allows this novel computational process is proposed as well as a physics-inspired mechanism that could explain its rather recent emergence without invoking unlikely genetic or structural changes. PMID- 16709405 TI - Glass transition and enthalpy relaxation of amorphous lactose glass. AB - The glass transition temperature, T(g), and enthalpy relaxation of amorphous lactose glass were investigated by differential scanning calorimetry (DSC) for isothermal aging periods at various temperatures (25, 60, 75, and 90 degrees C) below T(g). Both T(g) and enthalpy relaxation were found to increase with increasing aging time and temperature. The enthalpy relaxation increased approximately exponentially with aging time at a temperature (90 degrees C) close to T(g) (102 degrees C). There was no significant change observed in the enthalpy relaxation around room temperature (25 degrees C) over an aging period of 1month. The Kohlrausch-Williams-Watts (KWW) model was able to fit the experimental enthalpy relaxation data well. The relaxation distribution parameter (beta) was determined to be in the range 0.81-0.89. The enthalpy relaxation time constant (tau) increased with decreasing aging temperature. The observed enthalpy relaxation data showed that molecular mobility in amorphous lactose glass was higher at temperatures closer to T(g). Lactose glass was stable for a long time at 25 degrees C. These findings should be helpful for improving the processing and storage stability of amorphous lactose and lactose containing food and pharmaceutical products. PMID- 16709407 TI - Drosophila homologue of Eps15 is essential for synaptic vesicle recycling. AB - The mammalian protein Eps15 is phosphorylated by EGF receptor tyrosine kinase and has been shown to interact with several components of the endocytic machinery. We have identified a hypomorphic Eps15 mutant in Drosophila which shows reversible paralysis and an altered physiology at restrictive temperatures. In addition, the temperature-sensitive paralytic defect of shibire mutant is enhanced by this mutant. Eps15 is enriched in the larval neuromuscular junction in endocytic 'hot spots' in a pattern similar to Dynamin. Eps15 mutants show a decrease in the alpha-Adaptin levels at the larval neuromuscular junction synapse. Genetic and biochemical studies of interactions with components of the endocytic machinery suggest that Eps15 has an important role in synaptic vesicle recycling and regulates recruitment of alpha-Adaptin. PMID- 16709408 TI - Toxoplasma gondii: differential protection rates by two strains against cyst formation in a rat model. AB - A previous infection with the ME-49 strain of Toxoplasma gondii (of low pathogenicity for mice), protected 17 of 20 rats against formation of brain cysts, following challenge with 10(3) oocysts of the high pathogenicity M3 strain, as determined by bioassay of rat brains in mice. The low pathogenic KSU strain did not afford comparable protection. Protection was further tested in rats that were orally or subcutaneously immunized with cysts or oocysts of the ME 49 strain, and later challenged with 2 x 10(2) cysts or 10(2) oocysts of the highly pathogenic strains M3, M-7741 and C. Protection ranged from 43 to 100%, compared to non immunized control rats and was independent of the stage of ME-49 strain and of the routes used to immunize the rats. The results obtained encourage further investigation into prevention of toxoplasmosis in humans and food animals. PMID- 16709409 TI - Folding free-energy landscape of a 10-residue mini-protein, chignolin. AB - Chignolin is an artificial mini-protein composed of 10 residues (GYDPETGTWG) that has been shown to cooperatively fold into a beta-hairpin structure in water. We extensively explored the conformational space of chignolin using a 180-ns multicanonical molecular dynamics (MD) simulation and analyzed its folding free energy landscape. In the MD trajectory, we found structures that satisfy 99% of the experimental restraints and are quite close to the experimentally determined structures with C(alpha) root-mean-square-deviations of less than 0.5 Angstroms. These structures formed a large cluster in the conformational space with the largest probability of existence, agreeing well with the experiment. PMID- 16709410 TI - Brain alpha-dystroglycan displays unique glycoepitopes and preferential binding to laminin-10/11. AB - alpha-Dystroglycan was quantitatively enriched from mammalian brain based on its uniform reactivity with Vicia villosa agglutinin and resolved into sub populations possessing or lacking the sulfated glucuronic acid epitope recognized by monoclonal antibody HNK-1. We generated a new monoclonal antibody specific for a glycoepitope on brain alpha-dystroglycan but absent from alpha-dystroglycan expressed in all other tissues examined. Finally, we found that laminin-10/11 preferentially bound to brain alpha-dystroglycan compared to skeletal muscle alpha-dystroglycan. Our results suggest that tissue-specific glycosylation modifies the laminin binding specificity of alpha-dystroglycan. PMID- 16709411 TI - Upregulation of thromboxane synthase in human colorectal carcinoma and the cancer cell proliferation by thromboxane A2. AB - Tumor growth of colorectal cancers accompanies upregulation of cyclooxygenase-2, which catalyzes a conversion step from arachidonic acid to prostaglandin H(2) (PGH(2)). Here, we compared the expression levels of thromboxane synthase (TXS), which catalyzes the conversion of PGH(2) to thromboxane A(2) (TXA(2)), between human colorectal cancer tissue and its accompanying normal mucosa. It was found that TXS protein was consistently upregulated in the cancer tissues from different patients. TXS was also highly expressed in human colonic cancer cell lines. Depletion of TXS protein by the antisense oligonucleotide inhibited proliferation of the cancer cells. This inhibition was rescued by the direct addition of a stable analogue of TXA(2). The present results suggest that overexpression of TXS and subsequent excess production of TXA(2) in the cancer cells may be involved in the tumor growth of human colorectum. PMID- 16709412 TI - The neural cell adhesion molecule binds to fibroblast growth factor receptor 2. AB - The neural cell adhesion molecule (NCAM) can bind to and activate fibroblast growth factor receptor 1 (FGFR1). However, there are four major FGFR isoforms (FGFR1-FGFR4), and it is not known whether NCAM also interacts directly with the other three FGFR isoforms. In this study, we show by surface plasmon resonance analysis that NCAM can bind to FGFR2 with an affinity similar to that for the NCAM-FGFR1 interaction. However, the kinetic parameters for the NCAM-FGFR2 binding are different from those of the NCAM-FGFR1 binding. Both receptors were shown to cycle relatively fast between the NCAM bound and unbound states, although FGFR2 cycling was clearly faster (13 times) than the FGFR1 cycling. Moreover, ATP was more effective in inhibiting the binding of NCAM to FGFR1 than to FGFR2, indicating that the binding sites in NCAM for the two receptors are similar, but not identical. PMID- 16709413 TI - Role of the leader sequence in tobacco pectin methylesterase secretion. AB - We report that unprocessed tobacco pectin methylesterase (PME) contains N terminal pro-sequence including the transmembrane (TM) domain and spacer segment preceding the mature PME. The mature portion of PME was replaced by green fluorescent protein (GFP) gene and various deletion mutants of pro-sequence fused to GFP were cloned into binary vectors and agroinjected in Nicotiana benthamiana leaves. The PME pro-sequence delivered GFP to the cell wall (CW). We showed that a transient binding of PME TM domain to endoplasmic reticulum membranes occurs upon its transport to CW. The CW targeting was abolished by various deletions in the TM domain, i.e., anchor domain was essential for secretion of GFP to CW. By contrast, even entire deletion of the spacer segment had no influence on GFP targeting. PMID- 16709414 TI - Impact of organizational structure on nurses' job satisfaction: a questionnaire survey. AB - BACKGROUND: Nurses' job satisfaction is an important issue because of its impact on the quality of the nursing job. Therefore, it receives a lot of attention in the international nursing literature but insight into the sources of nurses' job satisfaction is yet insufficient, in particular for sources related to organizational structure. OBJECTIVES: We contribute by investigating the relationship between the organizational structure variables, formalization, centralization and specialization, and nurses' job satisfaction. This allows us to learn whether structural changes can help to improve satisfaction, and therefore nurses' work quality. DESIGN: Data were collected by questionnaires in a random sample of 764 non-managing nurses in three Belgian general care hospitals. We measured satisfaction by Stamps and Piedmont's work satisfaction index. Structure was also measured by an existing scale. RESULTS: The results support the negative effect of centralization and the clearly positive effects of specialization and formalization on nurses' job satisfaction. These effects differ according to the different dimensions of satisfaction. Furthermore, pay is the most important dimension of nurses job satisfaction but the dimension least influenced by organizational structure. CONCLUSIONS: The importance of the dimension pay in nurses' job satisfaction, which is not a function of organizational structure, is limiting hospitals in improving nurses' job satisfaction. However, organizational structure is related to the other dimensions of satisfaction. Especially, the fact that specialization and formalization are, contrary to our hypotheses, only positively related to satisfaction is important from a practice perspective and for further research. Furthermore, our data indicates that there is a need to refine one of the dimensions of Stamps and Piedmont index. PMID- 16709415 TI - Retention modelling in ternary solvent-strength gradient elution reversed-phase chromatography using 30 mm columns. AB - An optimization strategy for ternary solvent-strength gradient elution RP chromatography is described in which a two-dimensional model of gradient time (2 levels) against ternary proportions of organic modifiers (4 levels) was constructed. From the resolution surface the optimum ratio of organic modifiers could be selected. Excellent retention time and acceptable peak width and resolution simulations were obtained. The separation could be further optimized from the same input data by using a standard one-dimensional model in order to optimize for gradient slope, duration and shape. Excellent retention time and acceptable peak width and resolution simulations were obtained (< 1, 2 and 6% error, respectively). PMID- 16709417 TI - Separation of Am3+ and Eu3+ using an extraction chromatographic resin containing bis(2,4,4-trimethylpentyl)dithiophosphinic acid as the stationary phase. AB - Sorption of Am3+ and Eu3+ onto an extraction chromatographic resin material, prepared by impregnating purified bis(2,4,4-trimethylpentyl)dithiophosphinic acid (commercially available as Cyanex 301) into Chromosorb W, was investigated. Separation factor (S.F. = K(d,Am)/K(d,Eu)) values tend to increase in the presence of complexing agents such as thiocyanate and nitrate. In presence of 1 M NaNO3 in the aqueous phase, a S.F. value of approximately 1000 was obtained. The nature of extracted species was ascertained by carrying out the sorption studies at different aqueous phase pH. A column made from 300 mg of the resin material was used for the separation of Am3+ and Eu3+. A synthetic solution containing 1x10(5) cpm each of Am3+ and Eu3+ when loaded on to the column, >99% Eu was eluted out in 60 mL of 1 M NaNO3 at a pH of 3.2 while Am was eluted out using 5 mL of 1 M HNO3. The performance of the resin material was found to be reasonably good even after three cycles. PMID- 16709418 TI - Aspects of ionic diffusion through thick matrices of charged particles. AB - A review of the literature on ionic diffusion through matrices of charged particles shows that many workers have reported higher diffusivity of co-ions than of counterions. If these observations are correct then the requirement of electroneutrality of bulk solutions is violated. In those experiments other, not looked for, ions must have been taking part. It therefore appears that complete chemical analyses of solutions of both up- and downstream sides should be carried out, otherwise misleading inferences may be drawn, with practical consequences. Some researchers have studied transport of water under osmotic pressure differentials across clay membranes. However, a diffusing ion, during its transport from higher concentration to lower concentration, always carries its water of hydration. This second water transport process has not received attention. This suggests that at a critical concentration two transport processes will cancel each other. Attention has been drawn to this second type of water flow and its consequences. In ionic diffusion, each side of the membrane-outside solution interface is subject to the effects of the Nernst layer and Donnan zone. However, these layers and zones are seldom treated quantitatively. This has been carried out in this paper by two different methods. Both methods give identical thicknesses of the Nernst layer and Donnan zone. Finally, the effects of the presence of soluble bivalent salts on the diffusivity of different types of ions have been commented up on. In these cases some of the co-ions diffuse to the upstream side. PMID- 16709416 TI - Critical aspects of the determination of pentafluorobenzyl derivatives of aldehydes by gas chromatography with electron-capture or mass spectrometric detection: Validation of an optimized strategy for the determination of oxygen related odor-active aldehydes in wine. AB - This work presents a thorough study of some aspects critical to the quantitative performance of methods for the determination of volatile aldehydes previously derivatized to pentafluorobenzyl hydroxylamine oximes. The conclusions of the study are further applied to the validation of an optimized procedure for the determination of oxidation-related aldehydes from wine. Aspects studied include the chromatographic injection, the analytical performance of electron-capture detection (ECD) or MS detection, and the way in which the derivatization is carried out. Different injection techniques have been optimized and compared (classical splitless-hot injection; cold splitless; and large volume solvent split injection). All of them were carried out in a programmed temperature vaporization (PTV) injector, with a 350 microL internal volume liner (3.4 mm internal diameter). Classical splitless injection of hexane extracts is troublesome and requires large carrier gas flows (>10 mL min(-1)). Cold splitless injection was clearly superior. Large volume solvent split injection has been also optimized. ECD has been found to lack the necessary selectivity for the determination of unsaturated aldehydes in wine, although the quantitation of several minor aldehydes is possible. MS detection has proven to be superior for the determination of these compounds in wine. The way in which the reagent is applied is also critical and for the case of wine is important to ensure that the reagent is applied after some of the major wine aldehydes have been eliminated. The finally proposed method is extremely sensitive. Method detection limits ranged from 0.002 microg L(-1) (for unsaturated aldehydes) to 0.73 microg L(-1) (for phenylacetaldehyde) and precision (measured as relative standard deviation) is < or =6% in all cases. The method makes it possible to determine quantitatively in a single run the wine aldehydes with sensory significance (isobutyraldehyde, 2-methylbutanal, isovaleraldehyde, (E)-2-hexenal, (E)-2 heptenal, (E)-2-octenal, (E)-2-nonenal, methional and phenylacetaldehyde). PMID- 16709419 TI - Disseminated microsporidiosis due to Encephalitozoon hellem in an Egyptian fruit bat (Rousettus aegyptiacus). AB - Disseminated microsporidiosis was diagnosed in an adult female Egyptian fruit bat that died unexpectedly in a zoo. Gross findings, which were minimal, included poor body condition, bilateral renomegaly, and mottling of the liver. Histopathological lesions, which were particularly pronounced in the urogenital tract and liver, consisted primarily of inflammation associated with intracytoplasmic microsporidian spores. Polymerase chain reaction -based methods were used to establish the identity of the microsporidian as Encephalitozoon hellem. E. hellem is an emerging cause of human and avian disease, manifested mainly as opportunistic infection in immunosuppressed patients. This report describes the first documented case of E. hellem in a non-human mammalian species. PMID- 16709421 TI - Anomalous origin of the left coronary artery from the right side of the aortic valve in Syrian hamsters (Mesocricetus auratus). AB - This study describes the coronary artery distribution patterns associated with the anomalous origin of the left coronary artery from the right side of the aortic valve in Syrian hamsters. The hearts of 15 affected animals were examined by means of a corrosion-cast technique, histology and scanning electron microscopy. The hamsters belonged to a laboratory inbred colony with a high incidence of coronary artery anomalies and bicuspid aortic valves. The aortic valve was tricuspid in eight hamsters and bicuspid in the other seven. In all cases, the right coronary artery was normal, whereas the left main coronary artery trunk arose from the right aortic sinus or from the right side of the ventral aortic sinus when the aortic valve was bicuspid. In 12 specimens, the left main trunk crossed the infundibular septum and then divided into the left circumflex branch and the obtuse marginal branch. In another specimen, the course of the left main trunk was ventral to the right ventricular outflow tract; in the remaining two, it surrounded the aorta dorsally. In man, some of these distribution patterns may cause myocardial ischaemia and sudden death. The present findings prove that the origin of the left coronary artery from the right aortic sinus occurs in primitive mammals such as the Syrian hamster, suggesting that the defect may occur in other mammalian species. Its possible occurrence should be borne in mind in domestic animals, especially in those with signs of myocardial ischaemia after strenuous activity. PMID- 16709420 TI - Pancreatitis associated with hyperlipoproteinaemia type I in mink (Mustela vison): earliest detectable changes occur in mitochondria of exocrine cells. AB - Pancreatic tissue from young mink homozygous for a mutation in the lipoprotein lipase gene was studied by light and electron microscopy, with the aim of describing the earliest detectable changes in a process which rapidly progresses into overt pancreatitis. The mutation leads to hyperlipoproteinaemia, corresponding to hyperlipoproteinaemia type I in man. Assessment of relevant hepatic and pancreatic enzymes were included in the investigation. The earliest detectable changes consisted of widespread swelling and vacuolation of exocrine cells, arising mainly from swollen mitochondria. To a lesser extent, vesiculation of endoplasmic reticulum occurred. Mitochondria exhibited various changes, including cavitation and dilution of the matrix, with shortened and disorganized cristae displaced towards the periphery. Lamellar figures that developed within mitochondria were numerous. Acinar lumina were somewhat dilated, while plasma membranes were relatively well preserved and secretory granules seemed unchanged. Exfoliative processes progressively occurred, resulting in total necrosis of groups of parenchymal cells, while intercalated ducts were spared. The necrosis was rapidly followed by inflammatory reactions. The activity of the mitochondrial enzyme carnitine O-palmitoyltransferase, essential for the transport of fatty acids into the mitochondria, was lower in the pancreas than in the liver. The activity of the peroxisomal fatty acid beta-oxidation was high in the liver and low in the pancreas of both lipoprotein lipase-deficient and control mink. It is concluded that pancreatic lesions associated with hyperlipoproteinaemia start in exocrine cells, and are most probably the result of a metabolic disturbance, possibly a toxic effect of an excess of free fatty acids. PMID- 16709422 TI - Detection of maedi-visna virus in the kidneys of naturally infected sheep. AB - Infections with maedi-visna virus (MVV) cause progressive inflammation in different organs, mainly the lung, mammary gland, brain and joints. The aim of the present study was to investigate whether the kidney represents a viral target in natural MVV infection. For this, kidney samples from 13 sheep naturally infected with MVV were examined by histology, polymerase chain reaction (PCR), and immunohistochemistry. The kidneys of nine animals showed membranoproliferative glomerulonephritis and interstitial nephritis. The inflammatory infiltrate consisted of lymphocytes, plasma cells and macrophages. Interestingly, lymphoid follicles resembling those known to occur in other MVV infected tissues were observed. Lung tissue from the same animals had typical MVV lesions, such as lymphofollicular hyperplasia and interstitial pneumonia. Maedi visna proviral DNA sequences were detected in renal and lung tissue samples from these nine sheep by PCR, and the specificity of the amplified products was further verified by DNA sequencing. Moreover, MVV-specific immunohistochemistry revealed viral antigen in affected kidneys and lungs. These results suggest that the kidney may be a common target in natural MVV infection, and raise the issue of the role of this organ in the disease. PMID- 16709423 TI - Hydroponic uptake of atrazine and lambda-cyhalothrin in Juncus effusus and Ludwigia peploides. AB - Phytoremediation encompasses an array of plant-associated processes known to mitigate contaminants from soil, sediment, and water. Modification of pesticides associated with agricultural runoff includes processes directly associated with aquatic macrophytes in addition to changes in soil geochemistry and associated rhizospheric degradation. Remediation attributes of two vegetative species common to agricultural drainages in the Mississippi Delta, USA, were assessed using atrazine and lambda-cyhalothrin. Concentrations used in 8-d hydroponic exposures were calculated using recommended field applications and a 5% runoff model from a 0.65-cm rainfall event on a 2.02-ha field. While greater atrazine uptake was measured in Juncus effusus, greater lambda-cyhalothrin uptake occurred in Ludwigia peploides. Maximum pesticide uptake was reached within 48h for each exposure and subsequent translocation of pesticides to upper plant biomass occurred in macrophytes exposed to atrazine. Sequestration of 98.2% of lambda cyhalothrin in roots of L. peploides was measured after 8d. Translocation of lambda-cyhalothrin in J. effusus resulted in 25.4% of pesticide uptake partitioned to upper plant biomass. These individual macrophyte remediation studies measured species- and pesticide-specific uptake rates, indicating that seasonality of pesticide applications and macrophyte emergence might interact strongly to enhance mitigation capabilities in edge-of-field conveyance structures. PMID- 16709424 TI - Influences of macroalga-derived dissolved organic carbon on the aquatic toxicity of copper and cadmium. AB - In this study, the effect of dissolved organic carbon (DOC) derived from macroalga (Sargassum) on the acute toxicity of copper (Cu) and cadmium (Cd) to a freshwater cladoceran (Daphnia magna) was investigated. Potassium-loaded macroalga was incubated with ultrapure water to extract macroalgal DOC, which was then spiked with the constituents of the Elendt M7 hard water media. The 48 h median lethal concentration of Cu increased linearly with DOC levels but that of Cd was relatively independent of DOC levels (0-44 mg l(-1)). The independence of Cd toxicity on DOC level might be due to the competitive effect of high calcium concentrations in the media with Cd for the binding sites of DOC. The decreased Cu toxicity was a result of reduced Cu uptake as evidenced in a separate accumulation test. Also, the capability of the macroalgal DOC on reducing Cu toxicity was found to be comparable to DOC tested in other studies. Therefore, the present study suggested that the biosorption treatment process using macroalgae should consider the effect of DOC release from the biomass as a step of modifying the metal toxicity as well as influencing metal biosorption capacity. PMID- 16709425 TI - Evaluation of diffusive gradients in thin film (DGT) samplers for measuring contaminants in the Antarctic marine environment. AB - This work has been the first application of DGT samplers for measuring metals in water and sediment porewater in the Antarctic environment, and whilst DGT water sampling was restricted to quantification of Cd, Fe and Ni, preconcentration using Empore chelating disks provided results for an additional nine elements (Sn, Pb, Al, Cr, Mn, Co, Cu, Zn, As). Although higher concentrations were measured for some metals (Cd, Ni, Pb) using the Empore technique, most likely due to particulate-bound or colloidal species becoming entrapped in the Empore chelating disks, heavy metal concentrations in the impacted Brown Bay were found to be comparable with the non-impacted O'Brien Bay. Sediment porewater sampling using DGT also indicated little difference between Brown Bay and O'Brien Bay for many metals (Cd, Al, Cr, Co, Ni, Cu), however, greater amounts of Pb, Mn, Fe and As were accumulated in DGT probes deployed in Brown Bay compared with O'Brien Bay, and a higher accumulation of Sn was observed in Brown Bay inner than any of the other three sites sampled. Comparison of DGT derived porewater concentrations with actual porewater concentrations showed limited resupply of Cd, Pb, Al, Cr, Mn, Co, Ni, Cu, Zn and As from the solid phase to porewater, with these metals appearing to be strongly bound to the sediment, however, resupply of Fe and Sn was apparent. Based upon our observations here, we suggest that Sn, and to a lesser extent Pb, are critical contaminants. PMID- 16709426 TI - Residues of dioxins (PCDD/Fs) and PCBs in eggs, fat and livers of laying hens following consumption of contaminated feed. AB - Laying hens were fed with feed from the Belgian dioxin incident diluted ten-fold with non-contaminated feed, resulting in concentrations of 61 ngTEQkg(-1) PCDD/Fs, 23 ngTEQkg(-1) non-ortho PCBs, 116 ngTEQkg(-1) mono-ortho PCBs and 3.2 mgkg(-1) of the seven indicator PCBs. Following exposure for seven days, feed was replaced by non-contaminated feed for a period up to six weeks. Concentrations of PCDD/Fs in eggs showed a maximum of 214 pgTEQg(-1) fat after nine days and decreased to 44 pgTEQg(-1) after seven weeks. Dioxin concentrations in abdominal fat of chickens killed just after the last treatment, or after 1, 3 or 6 weeks on clean feed were 69, 84, 54 and 41 pgTEQg(-1) fat, respectively. Concentrations in livers decreased more rapidly, being 35, 7, 4 and 3 pgTEQg(-1) tissue, respectively. In both eggs and tissues, total TEQ concentrations were 3-4 times higher. Concentrations of the seven indicator PCBs in egg fat showed a stronger decrease with concentrations of 16.5 microgg(-1) at the peak (day 9) and 2.2 microgg(-1) after seven weeks. Corresponding concentrations in abdominal fat were, respectively, 4.6 and 2.6 microgg(-1) fat, and in livers 0.77 and 0.14 microgg(-1) tissue. The ratio of indicator PCBs to PCDD/Fs in feed was 52200. In eggs this ratio was initially higher (85000), but decreased towards 50000 after six weeks on clean feed. In abdominal fat the ratio varied between 49000 and 67000. In livers, the ratio was initially low (22000) but increased to 45000 towards the end of the study. It is concluded that the behaviour of PCDD/Fs and PCBs in laying hens is comparable and that the use of indicator PCBs appears to be a good alternative for PCDD/Fs but only in the case of co-exposure to both PCBs and PCDD/Fs, such as in incidents with PCB oil. PMID- 16709427 TI - Changes in the "stages of change" as outcome measures of a smoking cessation intervention: a randomized controlled trial. AB - OBJECTIVE: Employing changes in the "stages of change" as alternative outcome measures of a smoking cessation intervention. METHOD: RCT carried out between 2002 and 2004 in Germany with 485 daily cigarette smokers recruited from a general population survey. The intervention was conceptually based on the transtheoretical model, involving individualized feedback letters generated by computerized expert-system technology. RESULTS: Latent transition analysis was applied to test models for patterns of change, to evaluate stage of change distributions and transitions over time, and to compare the intervention and control group. The patterns of change across three time points could best be described by a model including stability, one-stage regressions, and one-to-four stage progressions. There were no significant differences between the intervention and control group in the stage of change distributions at each time point or in the amount of stage transitions over time. CONCLUSION: The efficacy of the intervention could not be demonstrated. Reasons may include oversampling of smokers who are interested in health-related topics but do not intend to quit, and power problems due to substantial attrition. PMID- 16709428 TI - The Prestige oil spill: a laboratory study about the toxicity of the water soluble fraction of the fuel oil. AB - The Prestige oil spill caused severe effects on the coastal fauna and flora due to direct contact of organisms with the fuel oil. However, the water soluble fraction (WSF) of the fuel oil can also provoke deleterious effects in the long term and even in regions not directly affected by the spill. Our objective was to determine the toxicity of the WSF using a battery of laboratory toxicity tests. To obtain a WSF in the laboratory, a sample of the spilled fuel was mixed with adequate medium, sonicated, agitated and filtered. No cytotoxic effects were detected in RTG-2 cells exposed to the WSF. In an algae growth inhibition test (OECD test guideline 201) the WSF did not affect the growth of Chlorella vulgaris. Furthermore, acute and reproductive toxicity tests (OECD test guideline 202) carried out using Daphnia magna did not indicate any deleterious effect of the WSF. In a bioassay designed in our laboratory, D. magna were fed with algae previously exposed to the fuel, but no toxic effects were detected. However, the WSF was able to induce a dose-dependent increase of ethoxyresorufin-O-deethylase activity in RTG-2 cells, indicating the presence of chemicals that could cause sub-lethal effects to organisms. After chemical analyses it was established that the final total quantity of polyaromatic hydrocarbons dissolved in medium was approximately 70 ng/ml. These low concentrations explain the observed lack of toxicity. PMID- 16709429 TI - Uptake, metabolism, mutant frequencies and mutational spectra in lambda transgenic medaka embryos exposed to benzo[alpha]pyrene dosed sediments. AB - The goal of this study was to provide data supporting the use of lambda transgenic medaka (Oryzias latipes) embryos to evaluate mutagens in sediments. Embryos incubated directly on sediments dosed with the reference mutagen, benzo[alpha]pyrene (BaP), were examined for BaP uptake and metabolism. Mutant frequency and mutational spectrum were assessed in the cII transgene recovered from adult medaka livers exposed as embryos. Embryos rapidly accumulated 14C-BaP and metabolized BaP to polar metabolites, indicating sediment-sorbed BaP is available for bioaccumulation and medaka embryos are capable of bioactivating this mutagen. Exposure of embryos to BaP dosed sediments significantly induced cII transgene mutant frequencies with mutations predominantly being in G:C base pairs, consistent with known mechanisms of BaP mutagenesis in transgenic mice and fish. PMID- 16709430 TI - Urogenital papilla feminization in male Pomatoschistus minutus from two estuaries in northwestern Iberian Peninsula. AB - Recently, male urogenital papilla feminization (UGPF) in the sand goby Pomatoschistus minutus was reported in several UK estuaries with high levels of estrogenic compounds. The fact that this species is also common in southern European estuaries, together with its life-cycle characteristics, prompted us to investigate P. minutus UGPF in the northwestern of Iberian Peninsula. Specimens of P. minutus were periodically sampled during 2004 at several locations in two estuaries (Minho and Lima). Evidence for UGPF was recorded in both estuaries, the highest incidence being observed in the Lima estuary (50%). Estrogen levels (17beta-estradiol and estrone) above 100 pg/l were observed in both estuaries, the higher concentrations being found in the Lima estuary. Overall, the results suggest a relationship between the feminization of male P. minutus urogenital papilla and the presence of estrogenic compounds. In the future, more in-depth studies are required in order to use P. minutus as a sentinel species for estrogenic compound monitoring in estuaries. PMID- 16709431 TI - An index of biotic integrity based on the summer polyhaline zooplankton community of the Chesapeake Bay. AB - A zooplankton index of biotic integrity was developed for the polyhaline waters of the Chesapeake Bay using data from a long-term environmental assessment program in which both zooplankton and water quality were regularly monitored. Summer (July to September) sampling events were classified as either coming from impaired or reference (least-impaired) conditions based on water quality conditions. Seventeen zooplankton community metrics were evaluated under these criteria and nine were chosen for a composite index. These were the Simpson diversity index, and abundance of barnacle larvae, rotifers, cladocerans, copepods, total mesozooplankton, and predators. The composite index of biotic integrity correctly classified about 94% of the impaired samples and about 82% of the reference samples. Average classification efficiency was 88%. This index appears to be an effective measure of eutrophication for the summer polyhaline waters of the Chesapeake Bay ecosystem. PMID- 16709432 TI - Effect of tributyltin on reproduction in Japanese whiting, Sillago japonica. AB - We examined the effect of tributyltin (TBT) on reproduction in the Japanese whiting, Sillago japonica. Mature fish were placed in indoor 500-L polyethylene tanks (five males and three females per tank) with a flow-through system and received dietary exposure to tributyltin oxide at concentrations of 2, 20, or 200 microg/g for 30 days during the active spawning period. Eggs spawned from the fish were collected daily, and the floating egg rate, larval deformity, hatchability, and viable hatch were monitored. TBT concentration in eggs of 200 microg/g group ranged from 85.0 to 159.6 ng/g in the evaluation period (days 5 30). In this period, the floating egg rate (83.2%), viable hatchability (82.2%), and total number of viable larvae (422,000 larvae per 100 g of female) were all significantly decreased in the 200-microg/g group compared with the control group (93.0%, 91.9%, and 709,000 larvae, respectively). The rate of deformity (2.6%) in the 200-microg/g group was about three times that in the control group (0.8%), although this difference was not significant. From these results, the lowest observed effect concentration of TBT in eggs on reproduction in Japanese whiting was estimated to be less than 159.6 ng/g-eggs. PMID- 16709433 TI - Toxicity of an engineered nanoparticle (fullerene, C60) in two aquatic species, Daphnia and fathead minnow. AB - Water-soluble fullerene (nC60) has been shown to induce lipid peroxidation (LPO) in brain of juvenile largemouth bass (LMB, Micropterus salmoides) [Oberdorster, E., 2004. Manufactured nanomaterials (fullerenes, c60) induce oxidative stress in brain of juvenile largemouth bass. Environ. Health Persp. 112, 1058-1062]; and upregulate genes related to the inflammatory response and metabolism, most notably CYP2K4 [. Nanotoxicology: an emerging discipline evolving from 116 studies of ultrafine particles. Environ. Health Persp. 113, 823-839]. The initial study in LMB was performed using tetrahydrofuran (THF)-solubilized nC60, although C60 can also be solubilized by stirring in water. The current study investigates differences in acute toxicity to Daphnia magna between THF-solubilized and water stirred-nC60 as a range-find for further assays in adult male fathead minnow (FHM, Pimephales promelas). The daphnia 48-h LC50 for THF-nC60 was at least one order of magnitude less (0.8 ppm) than that for water-stirred-nC60 (> 35 ppm). FHM were dosed with either 0.5 ppm of THF- or water-stirred-nC60 for 48 h. There was 100% mortality in the THF-nC60-exposed fish between 6 and 18 h, while the water-stirred-nC60-exposed fish showed no obvious physical effects after 48 h. Water-stirred-nC60 elevated LPO in brain, significantly increased LPO in gill, and significantly increased expression of CYP2 family isozymes in liver as compared to control fish. PMID- 16709434 TI - Environmental induction of CYP1A-, CYP2M1- and CYP2K1-like proteins in tropical fish species by produced formation water on the northwest shelf of Australia. AB - Normal operation of oil well platforms results in the discharge of "produced formation water" (PFW). The expression of CYP1A, CYP2M1- and 2K1-like proteins was examined for use as possible biomarkers of PFW exposure. A pilot study at the Harriet A production platform, on the Northwest Shelf of Australia, had indicated that PFW contamination possibly contributes to induction of CYP1A- and 2K1/2M1 like proteins in Gold-Spotted Trevally (Carangoides fulvoguttatus). In a subsequent caged fish study in the same location, Stripey seaperch (Lutjanus carponotatus) were caught at a clean site, then distributed to three caging sites: A (near-field), B (far-field) and C (a non-impacted reference site). Fish were sampled at time T = 0, 3 and 10 days. Significant increases of CYP1A, one CYP2K1- and two CYP2M1-like proteins were noted at Site A at T = 10. For another CYP2K1-like protein, a significant increase was observed at Site A only at T = 3. Prevailing winds changed between days 6 and 8 of sampling, moving the surface water plume directly west, possibly impacting in situ PFW exposure. The results indicate that tropical fish CYP1A protein is sensitive to PFW exposure. Importantly, statistically significant environmental induction of both CYP2M1- and CYP2K1-like proteins in tropical fish due to PFW exposure had not previously been described and CYP2 family induction may represent possible new biomarkers (other than CYP1A) of PFW exposure. In addition, the novel fraction-specific response of CYP2K-like proteins requires further verification but offers promise for improved monitoring of sub-lethal responses in marine organisms. (Supported by Australian Institute of Marine Science, Apache Energy Ltd. and the Environmental Toxicology Research Program at The University of Mississippi). PMID- 16709435 TI - Cloning and expression pattern of peroxisomal enzymes in the mussel Mytilus galloprovincialis and in the thicklip grey mullet Chelon labrosus: generation of new tools to study peroxisome proliferation. AB - Aquatic organisms living in coastal/estuarine areas show peroxisome proliferation after exposure to different environmentally relevant pollutants. In order to generate new tools to assess peroxisome proliferation in aquatic animals, peroxisomal enzymes were cloned using degenerate primers in the mussel Mytilus galloprovincialis and in the thicklip grey mullet Chelon labrosus. Fragments of catalase (CAT), thiolase (THIO), polyamine oxidase (POX) and xanthine oxidoreductase (XOR) were cloned and their expression pattern studied in different tissues by semi-quantitative RT-PCR. In mussels, CAT, THIO, POX and XOR were expressed in digestive gland, mantle and gills while in mullets CAT, THIO and POX were expressed in liver, spleen, brain, heart, muscle and gills. XOR was mainly expressed in liver and heart. Mature mullets showed the highest expression of peroxisomal enzymes in liver, spleen and brain, while in juveniles expression was mainly found in muscle tissues, liver and gills. Laboratory experiments of exposure to organic pollutants are being performed to study the usefulness of these tools to study peroxisome proliferation in pollution biomonitoring programmes. PMID- 16709436 TI - Autophagy: role in surviving environmental stress. AB - This conceptual paper addresses the role of lysosomal autophagy in cellular defence against oxidative stress. A hypothesis is proposed that autophagic removal of oxidatively damaged organelles and proteins provides a second tier of defence against oxidative stress. Age pigment or lipofuscin is a product of oxidative attack on proteins and lipids and can accumulate in lysosomes, where it can generate reactive oxygen species (ROS) and inhibit lysosomal function, resulting in autophagic failure. It is further hypothesised that repeated triggering of augmented autophagy can protectively minimise lipofuscin generation; and that animals living in fluctuating environments, where autophagy is repeatedly stimulated by natural stressors, will be generically more tolerant of pollutant stress. Data for resistance to pollutant stress is presented, together with evidence for a correlation between lysosomal stability and macrobenthic diversity. Finally, we speculate that organisms making up functional ecological assemblages in fluctuating environments, where up-regulation of autophagy should provide a selective advantage, may be pre-selected to be tolerant of pollutant-induced oxidative stress. PMID- 16709437 TI - The support of adenosine release from adenosine kinase deficient ES cells by silk substrates. AB - Adenosine kinase deficient (Adk-/-) embryonic stem cells (ESCs) encapsulated in synthetic polymers have previously been shown to provide therapeutic adenosine release and transient seizure suppression in epileptic rats. Here we explored the utility of biopolymer-substrates to promote long-term adenosine release from Adk /- ESCs. Three different substrates were studied: (1) type I collagen (Col-1), (2) silk-fibroin (SF), and (3) poly(L-ornithine) (PO) coated tissue culture plastic. Adk-/- or wild type (wt) ESC-derived glial precursor cells were seeded on the substrates and cultured either in proliferation medium containing growth factors or in differentiation medium devoid of growth factors. In proliferation medium cell proliferation was higher and metabolic activity lower on Col-1 and PO substrates as compared to SF. Cells from both genotypes readily differentiated into astrocytes after growth factor removal on all substrates. Adk-/- cells cultured on biopolymers released significantly more adenosine than their wt counterparts at all developmental stages. Adenosine release was similar on SF and PO substrates and the amounts released from Adk-/- cells (>20 ng/ml) were considered to be of therapeutic relevance. Taken together, these results suggest that silk matrices are particularly suitable biomaterials for ESC encapsulation and for the design of adenosine releasing bioincubators for the treatment of epilepsy. PMID- 16709438 TI - The effect of bariatric surgery on adipocytokines, renal parameters and other cardiovascular risk factors in severe and very severe obesity: 1-year follow-up. AB - AIMS: To evaluate the effect of weight loss after bariatric surgery (BS) on peripheral adipocytokines, renal parameters and other cardiovascular risk factors (CVRFs). METHODS: A total of 70 (41 women) extremely obese adults were prospectively studied before and 12 months after surgery. CONTROLS: 24 (15 women) normal-weight adults. Anthropometric, biochemical and renal parameters were recorded. RESULTS: Presurgery, adiponectin (ADPN) was lower, whereas leptin, insulin resistance, C-reactive protein, creatinine clearance and albuminuria were higher in patients than controls (P<0.001). All parameters improved postsurgery. Changes in ADPN correlated negatively with leptin, insulin resistance, albumin, C reactive protein, and creatinine clearance. Multiple regression analysis: using changes in ADPN as the dependent variable, only changes in insulin resistance (P=0.005) and albumin (P=0.019) were significant independent determinants for changes in ADPN. No statistical differences were found in relation to the degree of obesity. CONCLUSION: Patients changed to obesity type I after surgery. This implies a substantial improvement of CVRFs including ADPN, creatinine clearance and albuminuria. Changes in plasma ADPN correlated negatively with insulin resistance and with albuminemia but not with renal parameters. The lack of differences between different degrees of obesity suggests that the relationship between weight and CVRFs no longer exists when obesity becomes very extreme. PMID- 16709439 TI - The longitudinal effects of social support and hostility on depressive tendencies. AB - This 14-year longitudinal study examined the independent association between perceived social support and the 5-year progression of depressive tendencies while taking into account the potential effects of childhood/adolescent anger and later hostility. From the on-going population based study of "Cardiovascular Risk in Young Finns", 553 male and 860 female participants responded to a revised version of Beck's Depression Inventory in 1992 and 1997. Hostility and perceived social support were assessed by self-rated questionnaires in 1992. Childhood/adolescent anger was assessed by parent-reports in 1983. Our results showed that higher levels of perceived social support were associated with the decrease of depressive tendencies after 5 years and lower levels of depressive tendencies prospectively and after 5 years. This association persisted after adjusting for childhood/adolescent anger and later hostility. In addition, hostility was strongly related to the 5-year increase of depressive tendencies and higher levels of depressive tendencies. Social support may therefore be a long-term protective factor from depression irrespective of personality characteristics, such as hostility and anger. PMID- 16709441 TI - Interleukin-1beta regulation of N-type Ca2+ channels in cortical neurons. AB - Interleukin-1beta (IL-1beta) has been found to play an important role in various diseases in the central nervous system (CNS) and exhibit neuroprotective effects in some conditions. The transmitter release in brain is controlled by voltage gated Ca(2+) channels (VGCCs), predominantly N-type Ca(2+) channels (NCCs). Although both IL-1beta and NCCs are implicated regulating excitotoxicity and Ca(2+) homeostasis, it is not known whether IL-1beta modulates NCCs directly. In present study, we examined the effects of IL-1beta treatment (10 ng/ml, 24 h) on NCCs in cultured cortical neurons using patch-clamp recording and immunoblot assay. Our results showed that IL-1beta decreased NCC currents by approximately 50%, which made up 40% of the whole-cell Ca(2+) current demonstrated by omega conotoxin-GVIA, and also significantly downregulated the expression of NCC protein. The residual Ca(2+) currents except L-type Ca(2+) channel currents and NCC currents were not affected by IL-1beta. Our finding, IL-1beta inhibits the activity of NCC via suppressing NCC protein expression provides new insight into the neuroprotective role of IL-1beta in CNS. PMID- 16709442 TI - A joint-centred model accounts for movement curvature and spatial variability. AB - Hand reaching toward a visual target needs the central nervous system (CNS) to encode target location and initial limb posture. Once these sensory inputs are encoded within a common frame of reference, the motor system builds a motor command to drive the limb towards the target. In order to face the controversy concerning the variables (in task-space versus joint-space) the motor system may manipulate to build a motor command, we propose to compare recorded hand pointing movements to a task-space vector model and to a simplified joint-centred model. Subjects repeated large 3D free movements toward 12 different targets lying on a pointing table. Corresponding endpoint confidence ellipses were then computed. The main directions of these ellipses were finally compared to those predicted by the two models. The present work differs from earlier related studies in the fundamental approach data were recorded. Indeed, we put our emphasis on isolating the motor planning component from other online sensorimotor processes and higher level cognitive processes. In particular, we did not impose cognitively controlled movement features such as movement shape or planar constraint. Furthermore, a precise control of online feedbacks allowed us dissociating motor planning from online feedback processing. The obtained data better fitted the joint-centred model than the task-space vector model. Moreover, the recorded data exhibited curved trajectories very similar to the simulated values obtained from the joint-centred model. PMID- 16709443 TI - Induction of CB1 cannabinoid receptor by inflammation in primary afferent neurons facilitates antihyperalgesic effect of peripheral CB1 agonist. AB - Cannabinoids act on various regions in the nervous system to modulate neuronal activity including nociception. Here, we investigated CB1 receptor expression in primary afferent neurons in the dorsal root ganglion (DRG) and the efficacy of a local (intraplantar) application of the selective CB1 agonist, 2-arachidonyl-2 chloroethylamide (ACEA), on inflammatory thermal hyperalgesia. In situ hybridization showed normal CB1 mRNA expression in 28% of DRG neurons. Peripheral inflammation by CFA (complete Freund's adjuvant) significantly increased the ratio of CB1 mRNA-positive neurons to 43%, primarily with increase in NF200 negative C-fiber nociceptors. Furthermore, CB1 and TRPV1 (transient potential receptor vanilloid subtype-1) co-localization was increased from 41% before inflammation to 67% two days after inflammation. Inflammation also increased CB1 immunoreactivity in DRG neurons and in nerve fibers of the hindpaw dermis, indicating increased CB1 transport from the cell body to the peripheral nerve. The intraplantar application of ACEA attenuated CFA-induced thermal hyperalgesia. The antinociceptive properties of ACEA became more prominent at 2 days after inflammation, compared with those in non-inflamed and inflamed animals at 8 h. These results suggest that CB1 expression in primary afferent neurons is increased by inflammation and that the subsequent increase in CB1 transport to peripheral axons contributes to the increased antihyperalgesic efficacy of locally administered CB1 agonist. PMID- 16709440 TI - The enigmatic effects of caffeine in cell cycle and cancer. AB - Caffeine may very well be the most frequently ingested neuroactive drug in the world. Mechanistically, caffeine has been reported to affect cell cycle function, induce programmed cell death or apoptosis and perturb key cell cycle regulatory proteins. Although the effects of caffeine have been heavily investigated, much of the research data regarding caffeine's effects on cell cycle and proliferation seem ambiguous. One important factor may be that caffeine has been used experimentally in numerous cell types under a variety of conditions at concentrations ranging from micromolar to high millimolar. Physiologically, achieving experimental blood levels of caffeine would be extremely difficult without adverse side effects. Therefore, the relevance of experimental data obtained by using high concentrations of caffeine is not clear and may account for some of the discrepancies in the literature. This review attempts to reconcile data regarding the cellular effects of caffeine by examining reported effects on cell cycle, proliferation and apoptosis with careful attention to differences in experimental conditions and caffeine concentration utilized. PMID- 16709444 TI - Cholesterol is critical to the integrity of neuronal porosome/fusion pore. AB - Secretion is one of the most fundamental cellular processes. Porosomes have been demonstrated as the universal secretory machinery in cells. Earlier studies determine the presence of a number of proteins in porosomes, among them the N- and P/Q-type calcium channels, actin, syntaxin-1, synaptotagmin-1, vimentin, the N-ethylmaleimide-sensitive factor (NSF), the chloride channel CLC-3, and the alpha subunit of the heterotrimeric GTP-binding protein G(o). Studies demonstrate that t-SNAREs localize at the base of porosomes, and directly interact with calcium channels. In the present study, we demonstrate that Syntaxin-1 co localizes with cholesterol in solubilized synaptosomal membrane preparations. Depletion of cholesterol, results in the dissociation of both Syntaxin-1 and N type calcium channel from neuronal porosomes. Thus, cholesterol participates as an integral component of the neuronal porosome complex, and is required for its stability. PMID- 16709445 TI - Isolation and characterization of a Geobacillus thermoleovorans strain from an ultra-deep South African gold mine. AB - A thermophilic facultative bacterial isolate was recovered from 3.2km depth in a gold mine in South Africa. This isolate, designated GE-7, was cultivated from pH 8.0, 50 degrees C water from a dripping fracture near the top of an exploration tunnel. GE-7 grows optimally at 65 degrees C and pH 6.5 on a wide range of carbon substrates including cellobiose, hydrocarbons and lactate. In addition to O(2), GE-7 also utilizes nitrate as an electron acceptor. GE-7 is a long rod-shaped bacterium (4-6microm longx0.5microm wide) with terminal endospores and flagella. Phylogenetic analysis of GE-7 16S rDNA sequence revealed high sequence similarity with G. thermoleovorans DSM 5366(T) (99.6%), however, certain phenotypic characteristics of GE-7 were distinct from this and other previously described strains of G. thermoleovorans. PMID- 16709446 TI - General aspects of immunotoxicology including validation issues. AB - Histologic examination of lymphoid organs has revealed immunotoxic effects of a broad range of substances. The thymus has proven especially sensitive. The relative lack of sensitivity of mucosa-associated lymphoid cells and tissues may be due to shortcomings in the way they are examined. Validation of ways to examine mucosal lymphoid tissues and cells and development of histopathological tools to flag a compound as a potential inducer of autoimmune disease or allergy are challenges for the future. PMID- 16709447 TI - Histopathology and histomorphometry of the urogenital tract in 15-month old male and female rats treated neonatally with SERMs and estrogens. AB - In this study, two selective estrogen receptor modulators (SERMs), tamoxifen (TAM) and toremifene (TOR) or two estrogens, ethinylestradiol (EE) and diethylstilbestrol (DES) were administered to newborn male and female Sprague Dawley rats (days 1-5) to investigate the occurrence of developmental abnormalities in the adulthood. The compounds were dosed (s.c.) at an equimolar dose of 24.9 micromol/kg. During the follow-up period, mortality occurred mainly in DES-treated male rats (3/4), associated with obstructive urinary calculi and suppurative renal inflammation in 2/3 rats. Similar lesions were not evident in other groups. At the age of 15 months, the animals were necropsied and organs were collected for histopathology and histomorphometry. Treatment-related abnormalities were restricted to the reproductive organs. Chronic prostatitis and epithelial abnormalities in the vas deferens were observed in all treatment groups. The columnar epithelium of vas deferens showed hyperplasia and development of subepithelial glandular structures resembling epididymal cysts reported in humans exposed in utero to DES. Testicular atrophy was observed especially in estrogen-treated rats. Mainly in SERM-treated female rats, the uterus showed luminal dilation or obstruction, loss of endometrial glands and myometrium disorganization including foci of muscular disruption. TOR-treated female rats showed polyp-like nodules (incidence 4/15) and a high incidence (9/15) of a simple cuboidal epithelium in cervical regions normally occupied by multilayered epithelia. In conclusion, the vas deferens is a main target organ following neonatal administration of SERMs and estrogens. In addition, female rats were significantly more susceptible to SERM treatment than to treatment with estrogens. PMID- 16709448 TI - Design and evaluation of immunotoxicity studies. AB - The evaluation of potential adverse effects of pharmaceuticals on the immune system is part of the standard drug development procedures and needs to be available prior to the start of phase III clinical trials. Although the histopathological assessment of lymphoid organs/tissues is considered fundamental for the identification and characterization of immunotoxic reactions, additional investigations are now recommended by the European guidelines for repeated-dose toxicity testing of medicinal products in order to achieve an accurate assessment of immune system functionality with regard to immunomodulation. In the present paper, we describe and discuss a study design which permits the investigation of the immune function in a 4-week study in rats following immunization by subcutaneous administration of the T-dependent antigen Keyhole Limpet Hemocynin (KLH). This includes assessment of hematology parameters, titration of KLH specific antibodies in serum, lymphocyte immunophenotyping in blood, thymus, spleen and lymph nodes, macroscopic pathology and histopathological evaluation of the lymphatic organs/tissues and of the injection sites. PMID- 16709449 TI - The program for phenotyping of genetically modified animals at AstraZeneca. AB - Genetically modified mice offer a wide range of possibilities in preclinical drug discovery, e.g. for use in target identification, target validation and disease model generation. However, genomic modification and alteration in gene expression may cause unpredicted phenotypic alterations in the organism other than the intended ones. The aim of this study was to determine the importance of establishing the phenotype of transgenic and knockout mice models for use in pharmaceutical research. A total number of 51 mouse models (transgenic and knockout) produced at AstraZeneca during a 4 year period were subjected to a thorough phenotyping package covering clinical as well as morphological aspects. Phenotype abnormalities were recorded in 36 (70.6%) of the mouse models. The majority of findings were considered to be minor in magnitude. Histopathological changes related to the genotype of the animals were observed in 33% of the mouse models, underlining the importance of pathology in the phenotyping program. PMID- 16709450 TI - The effect of extracts from ginger rhizome on inflammatory mediator production. AB - Compounds from rhizomes of Zingiber officinale, commonly called ginger, have been purported to have anti-inflammatory actions. We have used an in vitro test system to test the anti-inflammatory activity of compounds isolated from ginger rhizome. U937 cells were differentiated and exposed to lipopolysaccharide (LPS) from Escherichia coli (1 microg/ml) in the presence or absence of organic extracts or standard compounds found in ginger (6-, 8-, 10-gingerol or 6-shogaol) for 24 h. Supernatants were collected and analyzed for the production of prostaglandin E(2) (PGE(2)) and tumor necrosis factor alpha (TNF-alpha) by standard ELISA assays. Predominant compounds in the organic extracts were identified as 6-, 8- 10 gingerols and 6-, 8-, 10-shogaols. Organic extracts or standards containing gingerols were not cytotoxic, while extracts or standards containing predominantly shogaols were cytotoxic at concentrations above 20 microg/ml. Crude organic extracts of ginger were capable of inhibiting LPS induced PGE(2) (IC(50)<0.1 microg/ml) production. However, extracts were not nearly as effective at inhibiting TNF-alpha (IC(50)>30 microg/ml). Thirty three fractions and subfractions, prepared by column chromatography, were analyzed for bioactivity. Extracts containing either predominantly gingerols or shogaols (identified by HPLC) were both highly active at inhibiting LPS-induced PGE(2) production (IC(50)<0.1 microg/ml), while extracts that contained unknown compounds were less effective (IC(50)<3.2 microg/ml). Extracts or standards containing predominantly gingerols were capable of inhibiting LPS-induced COX-2 expression while shogaol containing extracts had no effect on COX-2 expression. These data demonstrate that compounds found in ginger are capable of inhibiting PGE(2) production and that the compounds may act at several sites. PMID- 16709451 TI - Intestinal spasmolytic effects of STW 5 (Iberogast) and its components. AB - Functional gastro-intestinal diseases as the irritable bowel syndrome are very common in the population and are characterized by a broad spectrum of symptoms which mostly are related to spastic or paralytic intestinal function without defined histopathological changes of the tissue. Due to the multifactorial pathogenesis a multifactorial therapy with multi-target action seems to be reasonable. STW 5 (Iberogast), its constituent herbal extracts and some isolated compounds were used in an in vitro model provided by intestinal samples from guinea pig in order to test their activity on histamine-induced contractions and spontaneous motility, respectively. For comparison the known spasmolyticum papaverine was used. The results show that the lytic effect of the phytotherapeuticum on histamine-induced contraction represents additively the actions of the different components and corresponds to approx. 10 microM of papaverine. Spontaneous peristaltic motion was differently modulated by the various constituent extracts. The experiments with silibinin, glycyrrhicine, chelidonine, and protopine showed that the effects of the extracts were not comparable to those of the respective chemical constituents. PMID- 16709452 TI - Stenting allows weaning and extubation in ventilator- or tracheostomy dependency secondary to benign airway disease. AB - Central airway obstruction can cause severe respiratory insufficiency leading to mechanical ventilation (MV) or artificial airway (AA) dependency. Interventional bronchoscopic procedures have been reported to be of help in weaning patients with malignant airway stenoses from mechanical ventilation, whereas their use in benign disease is only anecdotal. The objectives of this study are to evaluate early, intermediate and long-term outcome of interventional bronchoscopy and stent placement in the treatment of MV/AA dependency due to benign airway obstruction. In a retrospective cohort analysis for the period 1999-2004, we evaluated 15 consecutive ICU patients with documented benign central airway obstruction, who were referred for bronchoscopic management of their condition after multiple failed attempts at weaning from MV or decannulation of the AA. Indications for bronchoscopic treatment were surgery refusal, medical or surgical inoperability, or absence of alternative treatment options. Malacia, post intubation stenosis and goiter were the main causes of airway obstruction and MV/AA dependency. All patients were treated by means of rigid bronchoscopy, dilatation procedures and stent insertion. All but one patient (93.3%) were successfully and permanently extubated/decanulated immediately after the bronchoscopy. Minor complications occurred in 6 patients (40%) leading to a second intervention. All complications could be managed endoscopically and long term follow up was uneventful. Interventional bronchoscopy with stent insertion can allow successful withdrawal from MV/AA and can offer longstanding airway patency in selected ventilator- or tracheostomy-dependant individuals with benign airway stenoses,when surgery in not feasable or contra-indicated. PMID- 16709453 TI - Binding of methoxy-substituted N1-benzenesulfonylindole analogs at human 5-HT6 serotonin receptors. AB - Comparison of several amine-substituted and methoxy-substituted analogs of N1-(4 aminobenzene)sulfonylindole suggests that these substituents might contribute to the 5-HT6 serotonin receptor affinity of these agents via their electronic effect on the indolic nucleus. Their 1,2,3,4-tetrahydrocarbazole counterparts behave differently. PMID- 16709454 TI - Discovery of thiochroman derivatives bearing a carboxy-containing side chain as orally active pure antiestrogens. AB - In order to search for alternatives to the sulfoxide moiety in the long side chain of pure antiestrogens, several molecules that may interact with water in a fashion similar to ICI164,384 were designed and it was found that compounds with the carboxy, the sulfamide, or the sulfonamide instead of the sulfoxide moiety also functioned as pure antiestrogens. Interestingly, the compound possessing the carboxy moiety showed superior antiestrogen activity compared to ICI182,780 when dosed orally. Results of the pharmacokinetic evaluation indicated that the potent antiestrogen activity at oral dosing attributed to both the improved absorption from the intestinal wall and the metabolic stability of the compound in liver. PMID- 16709455 TI - Magnetic resonance imaging of tumor cells by targeting the amino acid transport system. AB - An early diagnosis of cancer is crucial in the battle against this disease and the in vivo visualization of tumors at cellular level is still the most challenging goal. In order to target tumor cells, we took into account their increased metabolism and amino acid nutrients or pseudo-nutrients, which are actively transported through the cell membrane, have been chosen as vectors for new MRI contrast agents. For this reason new gadolinium complexes conjugated to agmatine, arginine, and glutamine have been synthesized and studied. PMID- 16709456 TI - Synthesis, structure analysis, and antitumor activity of 3,6-disubstituted-1,4 dihydro-1,2,4,5-tetrazine derivatives. AB - Fourteen compounds of 3,6-disubstituted-1,4-dihydro-1,2,4,5-tetrazine derivatives were prepared and their structures were confirmed by single-crystal X-ray diffraction and the semi-empirical calculation of PM3 method. This reaction yields the 1,4-dihydro derivatives rather than the 1,2-dihydro derivatives. The central six-membered ring of 1,4-dihydro-1,2,4,5-tetrazine has a chair conformation and therefore is not homoaromatic. Their antitumor activities were evaluated in vitro by SRB method for A-549 and BEL-7402 cells, and MTT method for P-388 and HL-60 cells. The results show that there is one compound which is highly effective against P-388 cells and one compound which is highly effective against HL-60 cells. So it is a kind of compound which possesses potential antitumor activities and is worth to research further. PMID- 16709457 TI - Improving anti-trypanosomal activity of 3-aminoquinoxaline-2-carbonitrile N1,N4 dioxide derivatives by complexation with vanadium. AB - New vanadium complexes of the type [V(IV)O(L)(2)], where L are 3-aminoquinoxaline 2-carbonitrile N(1),N(4)-dioxide derivatives, were prepared as an effort to obtain new anti-trypanosomal agents improving the bioactivity of the free ligands. Complexation to vanadium of the quinoxaline ligands leads to excellent antiprotozoal activity, similar to that of the reference drugs nifurtimox and benznidazole and in all cases higher than that of the corresponding free ligands. In addition, it is for the first time that the V((IV))O-quinoxaline complexes are reported as a family of anti-Trypanosoma cruzi agents. Finally, the anti trypanosomal activity of these vanadium complexes could be explained on the basis of their lipophilicity and the electronic characteristics of the quinoxaline substituents. PMID- 16709458 TI - Effects on erythroid differentiation of platinum(II) complexes of synthetic bile acid derivatives. AB - In this study, we compared some bile acid derivatives and their platinum(II) complexes with respect to their ability to induce erythroid differentiation of human leukemic K562 cells. The complexes analyzed were cis-[(3 dehydrocholanoyliden-L-tartrate)-diammineplatinum(II)] (compound 1) and cis [di(dehydrocholanoate)-bis(triphenylphosphine)-platinum(II)] (compound 3), together with their free ligands, respectively, 3-dehydrocholanoyliden-L-tartaric acid (compound 2) and dehydrocholanoic acid (4), and their parent compounds, respectively, cisplatin and cis-[dichloride-bis(triphenylphosphine)-platinum(II)] (5). We found that compound 1 stimulates erythroid differentiation of K562 cells and an increase of fetal hemoglobin (HbF) production in erythroid precursor cells isolated from peripheral blood of human subjects. This increase is similar to that obtained by hydroxyurea, a potent inducer of HbF production both in vitro and in vivo. Another important conclusion of this study is related to the evaluation of the effects of compound 1 on production of gamma-globin mRNA in human erythroid precursors grown in the two-stage liquid culture system. We demonstrated that compound 1 induces preferential accumulation of gamma-globin mRNA. The results presented in this manuscript could have practical impact, since it is well known that an increase in HbF production could ameliorate the clinical status of patients with beta-thalassemia and sickle cell anemia. PMID- 16709460 TI - Correlation of average muscle fiber conduction velocity measured during cycling exercise with myosin heavy chain composition, lactate threshold, and VO2max. AB - The aim of the study was to investigate the correlation between myosin heavy chain (MHC) composition, lactate threshold (LT), maximal oxygen uptake VO2max, and average muscle fiber conduction velocity (MFCV) measured from surface electromyographic (EMG) signals during cycling exercise. Ten healthy male subjects participated in the study. MHC isoforms were identified from a sample of the vastus lateralis muscle and characterized as type I, IIA, and IIX. At least three days after a measure of LT and VO2max, the subjects performed a 2-min cycling exercise at 90 revolutions per minute and power output corresponding to LT, during which surface EMG signals were recorded from the vastus lateralis muscle with an adhesive electrode array. MFCV and instantaneous mean power spectral frequency of the surface EMG were estimated at the maximal instantaneous knee angular speed. Output power corresponding to LT and VO2max were correlated with percentage of MHC I (R2=0.77; and 0.42, respectively; P<0.05). MFCV was positively correlated with percentage of MHC I, power corresponding to LT and to VO2max (R2=0.84; 0.74; 0.53, respectively; P<0.05). Instantaneous mean power spectral frequency was not correlated with any of these variables or with MFCV, thus questioning the use of surface EMG spectral analysis for indirect estimation of MFCV in dynamic contractions. PMID- 16709459 TI - Subcellular localization pattern of protoporphyrin IX is an important determinant for its photodynamic efficiency of human carcinoma and normal cell lines. AB - Photodynamic therapy (PDT) is a combination of light with a lesion-localizing photosensitizer or its precursor to destroy the lesion tissue. PDT has recently become an established modality for several malignant and non-malignant conditions, but it can be further improved through a better understanding of the determinants affecting its therapeutic efficiency. In the present investigation, protoporphyrin IX (PpIX), an efficient photosensitizer either endogenously induced by 5-aminolevulinic acid (ALA) or exogenously administered, was used to correlate its subcellular localization pattern with photodynamic efficiency of human oesophageal carcinoma (KYSE-450, KYSE-70) and normal (Het-1A) cell lines. By means of fluorescence microscopy ALA-induced PpIX was initially localized in the mitochondria, whereas exogenous PpIX was mainly distributed in cell membranes. At a similar amount of cellular PpIX PDT with ALA was significantly more efficient than photodynamic treatment with exogenous PpIX at killing all the 3 cell lines. Measurements of mitochondrial membrane potential and intracellular ATP content, and electron microscopy showed that the mitochondria were initially targeted by ALA-PDT, consistent with intracellular localization pattern of ALA induced endogenous PpIX. This indicates that subcellular localization pattern of PpIX is an important determinant for its PDT efficiency in the 3 cell lines. Our finding suggests that future new photosensitizers with mitochondrially localizing properties may be designed for effective PDT. PMID- 16709461 TI - HACA, the transcriptional activator of the unfolded protein response (UPR) in Aspergillus niger, binds to partly palindromic UPR elements of the consensus sequence 5'-CAN(G/A)NTGT/GCCT-3'. AB - The promoters of UPR target genes contain an unfolded protein response element (UPRE), which confers the stress inducibility to the gene, via an interaction with the transcription activator HACA. In the promoters of the Aspergillus ER stress responsive genes bipA, cypB, pdiA, prpA, tigA, and hacA, a consensus sequence was identified, which was located close to the transcription start site of the gene (<81 bp), and corresponds to the sequence CAN(G/A)NTGT/GCCT. The UPRE is a partly palindromic sequence around a dispensable spacer nucleotide, followed by four highly conserved bases. By an in vitro selection procedure, an optimal binding site for HACA was isolated. This sequence, ACACGTGTCCT, resembles the UPRE but lacks the spacer nucleotide. It has a much higher binding affinity than the identified UPREs, and in vivo it behaves as a more powerful cis-acting element. PMID- 16709462 TI - ELISE NMR: experimental liquid sealing of NMR samples. AB - We present a simple, generally applicable approach to prevent sample evaporation when working at elevated temperatures in high resolution NMR. It consists of experimentally sealing the NMR sample by a second liquid (Experimental Liquid Sealing, ELISE). For aqueous samples, we identified the mineral oil commonly used in PCR application as the best candidate, because it contains only a very limited amount of water-soluble contaminants, is stable over time and heat resistant. The procedure does not interfere with shim settings, and is compatible with a wide variety of samples, including oligosaccharides and proteins. For chloroform samples, a simple drop of water allows to efficiently seal the sample, avoiding solvent evaporation even over lengthy time periods. PMID- 16709463 TI - A comprehensive shape model of the heart. AB - Domain knowledge about the geometrical properties of cardiac structures is an important ingredient for the segmentation of these structures in medical images or for the simulation of cardiac physiology. So far, a strong focus was put on the left ventricle due to its importance for the general pumping performance of the heart and related functional indices. However, other cardiac structures are of similar importance, e.g., the coronary arteries with respect to diagnosis and treatment of arteriosclerosis or the left atrium with respect to the treatment of atrial fibrillation. In this paper we describe the generation of a geometric cardiac model including the four cardiac chambers and the trunks of the connected vasculature, as well as the coronary arteries and a set of cardiac landmarks. A mean geometric model for the end-diastolic heart has been built based on 27 cardiac CT datasets and has been evaluated with respect to its capability to estimate the position of cardiac structures. Allowing a similarity transformation to adapt the model to image data, cardiac surface positions can be predicted with an accuracy of below 5mm. PMID- 16709465 TI - Meeting the information needs of women with breast cancer: Piloting a nurse-led intervention. AB - This study evaluated a telephone intervention, administered by specialist breast care nurses, that aimed to meet the information needs of women with breast cancer. The intervention was developed from previous work that examined priority information needs. Participants were allocated to a telephone intervention (n = 67) or control group (n = 68). Data were collected by means of semi-structured interviews with participants at two time points (3 months and 8-12 months post diagnosis) and focused on patient satisfaction with sources of information, information needs and psychological morbidity. Interviews were also conducted with breast care nurses to ascertain their views on administering the intervention. The intervention group reported fewer physical problems at Time 2 and were more likely to have had their information needs met than women in the control group. The control group were more likely to utilise media sources of information at Time 2 whereas women in the intervention group reported breast care nurses as their most prominent source of information. There were no significant differences in psychological morbidity between the two study groups. The findings indicate that the intervention is a feasible and acceptable approach to meeting the information needs of women with breast cancer. Further research is required to evaluate the intervention in a randomised controlled trial. PMID- 16709464 TI - Effects of ramelteon on patient-reported sleep latency in older adults with chronic insomnia. AB - BACKGROUND AND PURPOSE: To assess the efficacy and safety of ramelteon, a selective MT(1)/MT(2) receptor agonist, for chronic insomnia treatment. PATIENTS AND METHODS: Randomized, double-blind, placebo-controlled 35-night outpatient trial with weekly clinic visits at multiple centers. Patients include older adults (>or=65 years; N=829) with chronic insomnia. Placebo, ramelteon 4mg, or ramelteon 8mg were taken nightly for five weeks, and patient-reported sleep data were collected using sleep diaries. Primary efficacy was sleep latency at week 1. Sustained efficacy was examined at weeks 3 and 5. Rebound insomnia and withdrawal effects were evaluated during a 7-day placebo run-out. RESULTS: Both doses of ramelteon produced statistically significant reductions in sleep latency vs. placebo at week 1 (ramelteon 4mg: 70.2 vs. 78.5min, P=.008; ramelteon 8mg: 70.2 vs. 78.5 min, P=.008). Patients continued to report reduced sleep latency at week 3 with ramelteon 8mg (60.3 vs. 69.3min, P=.003), and at week 5 with ramelteon 4 mg (63.4 vs. 70.6 min, P=.028) and ramelteon 8 mg (57.7 vs. 70.6 min; P<.001). Statistically significant increases in total sleep time were observed with ramelteon 4 mg at week 1 (324.6 vs. 313.9 min, P=.004) and week 3 (336.0 vs. 324.3min, P=.007) compared with placebo. There was no evidence of significant rebound insomnia or withdrawal effects following treatment discontinuation. The incidence of adverse events was similar among all treatment groups; most were mild or moderate. CONCLUSIONS: In older adults with chronic insomnia, ramelteon significantly reduced patient reports of sleep latency over five weeks of treatment with no significant rebound insomnia or withdrawal effects. PMID- 16709466 TI - New technologies for the control of human hookworm infection. AB - Since the 1990s, the major approach to hookworm control has been morbidity reduction in school-aged children by periodic deworming with benzimidazoles. Now, efforts are underway to determine the feasibility of integrating deworming with control programs that target other neglected tropical diseases. However, the sustainability of benzimidazole deworming for hookworm is of concern because of the variable efficacy of mebendazole, high rates of post-treatment reinfection and possible development of drug resistance. This requires parallel efforts to develop new and complementary hookworm control tools, such as new anthelmintic drugs (e.g. tribendimidine) and a recombinant hookworm vaccine. It is hoped that, ultimately, anthelmintic vaccination will be linked to deworming as part of an expanded control package. PMID- 16709467 TI - First LC-MS/MS electrospray ionization validated method for the quantification of perindopril and its metabolite perindoprilat in human plasma and its application to bioequivalence study. AB - A high throughput bioanalytical method based on solid phase extraction and liquid chromatography-tandem mass spectrometry (LC-MS/MS), has been developed for the estimation of perindopril and its metabolite perindoprilat, an angiotensin converting enzyme inhibitor in human plasma. Ramipril was used as internal standard (IS). The extraction of perindopril, perindoprilat and ramipril from the plasma involved treatment with phosphoric acid followed by solid phase extraction (SPE) using hydrophilic lipophilic balance HLB cartridge. The SPE eluate without drying were analyzed by LC-MS/MS, equipped with turbo ion spray (TIS) source, operating in the negative ion and selective reaction monitoring (SRM) acquisition mode to quantify perindopril and perindoprilat in human plasma. The total chromatographic run time was 1.5 min with retention time for perindopril, perindoprilat and ramipril at 0.33, 0.35 and 0.30 min. The developed method was validated in human plasma matrix, with a sensitivity of 0.5 ng/ml (CV, 7.67%) for perindopril and 0.3 ng/ml (CV, 4.94%) for perindoprilat. This method was extensively validated for its accuracy, precision, recovery, stability studies and matrix effect especially because the pattern of elution of all the analytes appears as flow injection elution. Sample preparation by this method yielded extremely clean extracts with very good and consistent mean recoveries; 78.29% for perindopril, 76.32% for perindoprilat and 77.72% for IS. The response of the LC-MS/MS method for perindopril and perindoprilat was linear over the range 0.5 350.0 ng/ml for perindopril and 0.3-40 ng/ml for perindoprilat with correlation coefficient, r>/=0.9998 and 0.9996, respectively. The method was successfully applied for bioequivalence studies in human subjects samples with 4 mg immediate release (IR) formulations. PMID- 16709468 TI - Enhanced extracellular matrix production and differentiation of human embryonic germ cell derivatives in biodegradable poly(epsilon-caprolactone-co-ethyl ethylene phosphate) scaffold. AB - Extracellular environment regulates cell behavior and also influences the differentiation of stem cells. Two cell lines of pluriopotent human embryonic germ cell derivatives (EBD cells) were cultured on a biodegradable poly(epsilon caprolactone-co-ethyl ethylene phosphate) (PCLEEP) and non-degradable cellulose acetate scaffold. Their cell behaviors including proliferation, differentiation, cell distribution and extracellular matrix production were studied for 4 weeks and 10 months. The proliferation of the EBD cells was enhanced in both of the three-dimensional scaffolds in the first 5 weeks of culture, regardless of the material difference, compared to monolayer culture. While the gene expression profile remained multilineage for the EBD cells cultured in the cellulose acetate fibrous scaffold, much of the neuronal lineage markers were down-regulated in EBD cells cultured in the PCLEEP scaffold. On the other hand, extracellular matrix production was significantly enhanced in the PCLEEP scaffold. The study showed that the polymer substrate could influence the differentiation and growth of pluripotent stem cells in the absence of exogenous biochemical signals. PMID- 16709470 TI - Introduction: women's occupational and environmental health. PMID- 16709469 TI - Correlates of bone quality in older persons. AB - PURPOSE OF THE STUDY: In a population-based sample of older persons, we studied the relationship between tibial bone density and geometry and factors potentially affecting osteoporosis. METHODS: Of the 1260 participants aged 65 years or older eligible for the InCHIANTI study, 1155 received an interview and 915 (79.2%) had complete data on tibial QCT scans and other variables used in the analysis presented here. The final study population included 807 persons (372 men and 435 women, age range 65-96 years) after exclusion of participants affected by bone diseases or treated with drugs that interfere with bone metabolism. RESULTS: In both sexes, calf cross-sectional muscle area (CSMA) was significantly and independently associated with total bone cross-sectional area (tCSA) and cortical bone cross-sectional area (cCSA) but not with trabecular or cortical volumetric bone mineral density (vBMD). Bioavailable testosterone (Bio-T) was independently associated with both trabecular and cortical vBMD in both sexes. In women, independently of confounders, 25(OH)-vitamin D was positively associated with tCSA and cortical vBMD, while PTH was negatively associated with cortical vBMD. IL-1 beta was negatively correlated with cortical vBMD in women, while TNF-alpha was associated with enhanced bone geometrical adaptation in men. CONCLUSIONS: Physiological parameters that are generically considered risk factors for osteoporosis were associated with specific bone parameters assessed by tibial QCT. Factors known to be associated with increased bone reabsorption, such as 25(OH)-vitamin D, PTH and Bio-T, affected mainly volumetric BMD, while factors associated with bone mechanical stimulation, such as CSMA, affected primarily bone geometry. Our results also suggested that pro-inflammatory cytokines might be considered as markers of bone resorption. PMID- 16709471 TI - Sex, gender and women's occupational health: the importance of considering mechanism. AB - A number of researchers have pointed out that less is known about occupational determinants of health in women than in men. The authors examine inventories of ongoing Canadian research and of recent scientific publications in order to identify trends in the approaches used to study women's occupational health (WOH). We also consider conceptual issues in the treatment of the sex and gender of subjects. We observe that women have been the subject of relatively few investigations of occupational health in the natural or biomedical sciences and that studies of WOH have concentrated on the health care professions and on psychosocial stressors, with a deficit in toxicological and physiological studies. We use recent studies of mercury exposure in chloralkali process plants and of musculoskeletal disorders among office workers to provide specific examples of problems in conceptualizing WOH. We propose that WOH be studied more often, especially by researchers in the natural and biomedical sciences, and that such studies include both women and men, where possible, and consider the complex relationships of gender and sex to the pathways involved. More interdisciplinary research would facilitate this process, since social researchers have tended to focus more on gender/sex issues. Our findings demonstrate that it is necessary to explore the implications of using sex routinely as an explanatory variable in occupational health research and to increase emphasis on the mechanisms involved in any sex or gender differences sought or found. From an equity perspective, it is also important to situate biological sex differences so as to prevent them from being used erroneously to justify job segregation or inequitable health promotion measures. PMID- 16709472 TI - Gender differences in respiratory symptoms-does occupation matter? AB - Little attention has been given to gender differences in respiratory health, particularly in occupational settings. The purpose of this paper was to evaluate gender differences in respiratory morbidity based on surveys of hospitality workers, radiographers, and respiratory therapists. Data were available from mail surveys of 850 hospitality industry workers (participation rate 73.9%; 52.6% female), 586 radiographers (participation rate 63.6%; 85% female), and 275 respiratory therapists (participation rate 64.1%; 58.6% female). Cross tabulations by gender were evaluated by chi(2) analysis and logistic regression with adjustment for personal and work characteristics. Women consistently had greater respiratory morbidity for symptoms associated with shortness of breath, whereas men usually had a higher prevalence of phlegm. There were few differences in work exposures apart from perception of exposure to ETS among hospitality workers. Gender differences in symptoms were often reduced after adjustment for personal and work characteristics but for respiratory therapists there were even greater gender disparities for asthma attack and breathing trouble. Population health findings of elevated symptoms among women were only partially supported by these occupational respiratory health surveys. The influence of differential exposures and personal factors should be considered when interpreting gender differences in health outcomes. PMID- 16709473 TI - Epidemiology of venous thromboembolism in neonates and children. AB - Venous thromboembolism is an increasingly recognised problem in paediatric practice, particularly in the context of tertiary care paediatric services. In recent years, several national and international registries have helped to define the epidemiology of venous thromboembolism in both neonates and older children. These studies have generated information on the incidence and risk factors associated with venous thromboembolism in different age groups. Data from these and other studies have demonstrated important differences between paediatric and adult practice and highlight the need for specific evidence based guidelines for the prevention and management of venous thromboembolism in neonates and children. PMID- 16709474 TI - Perinatal and neonatal ischaemic stroke: a review. PMID- 16709475 TI - Hyperlipidemia in children. AB - Hyperlipidemia in children has emerged as an increasingly prevalent risk factor in children, concomitant with the worldwide epidemic of obesity. Hyperlipidemia can alter vascular endothelial function and impair some of its pro-fibrinolytic and anti-thrombotic regulatory properties, as well as initiate the atherosclerotic process. There are strong links between vascular changes and hyperlipidemia in children, both from pathologic and non-invasive assessment studies. More severe lipid abnormalities in children are related to primary familial dyslipidemias. Current recommendations for screening begin with assessment of family history for cardiovascular disease or events or parental hyperlipidemia. High-risk individuals merit more intensive investigation and intervention. While fat-restricted diets have been shown to be safe in children, lipid-lowering is modest. Those with more severe lipid abnormalities may meet criteria for drug therapy, and the statin agents commonly used in adults are increasingly being used in high-risk children, with similar efficacy and safety, although long-term concerns remain. PMID- 16709476 TI - Antiplatelet therapy in children. AB - Platelets are essential for the maintenance of vascular integrity and control of bleeding at sites of injury, but they are also implicated in the progression of atherosclerotic lesions and arterial vascular thrombosis. The use of antiplatelet drugs for the primary and secondary prevention of cardiovascular and cerebrovascular thromboses in adult populations has been extensively evaluated, resulting in defined management strategies. Much less is known about the appropriate use of antiplatelet drugs (primarily aspirin) in infants and children for secondary prevention in ischemic stroke, for prevention of coronary artery thrombosis in Kawasaki disease, or for prevention of thromboembolism following surgery for congenital cardiac disease. Additional studies will be required to evaluate the relative benefits of aspirin and anticoagulants in these settings. A role for newer antiplatelet drugs in the management of pediatric arterial thrombosis is as yet unexplored. PMID- 16709477 TI - Oral anticoagulation therapy in children. AB - Treatment of thromboembolic complications in children has been the subject of considerable research in the last decade. Recommendations for oral anticoagulant therapy in children have been extrapolated from adult clinical trials. Coumarin derivatives are the preeminent oral antithrombotic agents used in children. Warfarin, acenocoumarol and phenprocoumon are the vitamin K antagonists used in children with thrombotic complications in different countries according to their experience and familiarity within a country or region. Prospective studies from Canada and Argentina propose guidelines for administering and monitoring warfarin and acenocoumarol therapy in children. These studies highlight the difficulty of their use in pediatric patients. Infants younger than 12 months of age require increased doses to achieve and maintain the therapeutic target INR, adjustments of loading dose to achieve the target INR faster with no overshooting, more frequent INR testing and dose adjustments, and fewer INR in the target range. The current indications for oral anticoagulants in children with thrombotic complications, the side effects of these agents and the reversal of the anticoagulant effect are discussed. PMID- 16709478 TI - Thrombolytic therapy in children. AB - Thrombolysis is increasingly considered a treatment option in newborns and children with arterial and venous thromboembolic events, or occluded central venous lines. However, no uniform recommendations are available with regard to indications, drug of choice, route of administration, and dosing regimen. Thus, several protocols are used for the different thrombolytic agents, leading to differing outcome with respect to the effectiveness of therapy and bleeding complications. This article will summarize the available information on the use of thrombolytic agents in newborns and children, focussing on the potential indications, efficacy and safety profiles, and evidence supporting dosing schedules. PMID- 16709479 TI - Antithrombin replacement in neonates: is there any indication? AB - INTRODUCTION: Activation of the coagulation system and severe acquired antithrombin (AT) deficiency are common and prognostically important findings in sick and preterm neonates. It has been hypothesised that treatment of the acquired AT deficiency with AT concentrate may improve the outcome of conditions such as the neonatal respiratory distress syndrome (RDS), intracranial hemorrhage (ICH) and sepsis. MATERIALS AND METHODS: We performed a systematic review of randomised controlled trials (RCTs) of AT replacement therapy in newborn infants. RESULTS: Two full-length trial reports were found. Both were placebo-controlled. The first RCT examined the effects of AT therapy in 122 preterm infants with RDS. Administration of AT prolonged rather than shortened the duration of mechanical ventilation and oxygen therapy. The second RCT determined whether AT replacement decreased the incidence of ICH in 60 preterm infants who were born before 30 weeks of gestation. No beneficial effect on ICH was found. CONCLUSIONS: Preterm infants with RDS do not benefit from therapy with AT concentrate and may be harmed. There is also little evidence that the administration of AT reduces the risk of ICH. The role of AT replacement during neonatal sepsis remains uncertain. PMID- 16709480 TI - Point-of-care antithrombotic monitoring in children. AB - INTRODUCTION: The use of oral anticoagulant therapy is increasing in children. Managing anticoagulant therapy in children presents unique challenges, including poor venous access. The advent of point-of-care (POC) monitoring of anticoagulant therapy offers a potential solution to this challenge. This paper reviews the published literature relating to POC monitoring of oral anticoagulant therapy in children. MATERIALS AND METHODS: A Medline search was conducted and identified key publications. Papers were reviewed with respect to their objectives, populations and POC device investigated. Study limitations were identified. RESULTS: Five publications and one abstract were identified, reporting studies using five different POC monitors. Three studies had a strong clinical management focus. Outcome measures assessed included target therapeutic range achievement and frequency of adverse events. Correlation between POC and laboratory-based results ranged from 0.83 to 0.96. Home monitoring and self-management using POC monitors were both reported to be preferred compared to standard laboratory testing. CONCLUSIONS: POC monitoring of oral anticoagulant therapy in children offers considerable advantages. The reviewed literature would suggest such monitoring can be performed accurately and reliably. The impact of quality control issues, such as calibration of thromboplastin ISI in POC devices, has not been explored in a paediatric population. Further studies are needed to clarify such issues and confirm the safety, reliability and efficacy of POC monitoring of oral anticoagulant therapy in children, including its home monitoring and self management programs. PMID- 16709481 TI - Diagnosis and treatment of heparin-induced thrombocytopenia in neonates and children. AB - Heparin-induced thrombocytopenia (HIT), a well-known side effect of heparin therapy, occurs with an incidence of 1-2% in certain pediatric patient groups. In affected children, HIT markedly increases the risk of venous and arterial thromboembolism. The use of alternative anticoagulation with danaparoid, lepirudin and argatroban in adults and children has demonstrated to be safe and could reduce morbidity and mortality also in affected pediatric patients. Thus, in children and neonates, an early diagnosis and accurate management is crucial to avoid the deleterious consequences of HIT. This review article will focus on the presentation of HIT in neonates and children. It reviews the pathophysiology of HIT and it summarizes epidemiological data. Finally important diagnostic and therapeutic issues are discussed. PMID- 16709482 TI - Spitz nevus versus melanoma: limitation of the diagnostic methodology exposed. PMID- 16709483 TI - Highlighting the rim of the perifollicular epidermal unit. AB - The perifollicular and interfollicular areas of normal skin may look similar. However, some physiological and pathological processes may specifically involve a thin perifollicular rim. This review illustrates some of the methods available for highlighting the rim of the perifollicular epidermal unit. Non invasive methods rely on dermoscopy, ultraviolet light enhanced visualization (ULEV), skin capacitance imaging and cyanoacrylate skin surface strippings (CSSS). Conventional histology and immunohistochemistry may also show specific perifollicular features without, however, revealing the aspects highlighted by the specific non invasive methods. The clinically relevant modifications consist of pigmentary and hyperkeratotic perifollicular changes. PMID- 16709484 TI - The porphyrias: clinical presentation, diagnosis and treatment. AB - The porphyrias comprise a clinically and genetically heterogeneous group of diseases mostly arising from a genetically determined dysfunction of specific enzymes along the pathway of heme biosynthesis. Based on the occurrence or absence of cutaneous symptoms and life-threatening acute neurological attacks, the different types of porphyrias can either be classified into cutaneous and non cutaneous forms or acute and non-acute forms. Establishing an accurate diagnosis might be difficult for two reasons: i) the porphyrias can manifest with a broad but unspecific spectrum of clinical symptoms mimicking several other disorders, and ii) biochemical examination of urine, feces, and blood can reveal overlapping findings. Fortunately, however, the advances in the fields of molecular genetics during recent years have provided us with the possibility of overcoming these diagnostic pitfalls. Therefore, in controversial cases the correct diagnosis can finally be made using molecular biological techniques. Due to the various facets of the porphyrias, diagnosis and treatment should always imply a close interdisciplinary collaboration to counsel and help patients and their families most efficiently. PMID- 16709486 TI - Linkage of a locus determining familial progressive hyperpigmentation (FPH) to chromosome 19p13.1-pter in a Chinese family. AB - Familial progressive hyperpigmentation (FPH) is a rare autosomal dominantly inherited disorder characterized by patches of hyperpigmentation in the skin which are present at birth or in early infancy and increase in size and number with age. Although previous studies showed that FPH is a monogenic trait, the genetic basis for this disease is unknown. Using a genome screening with 182 STR markers from autosomes in a three-generation Chinese family with 17 members, including 6 affected individuals, we identified a locus linked to chromosome 19p13.1-pter responsible for FPH, spanning 45.48 cM between D19S593 and 19pter. Interestingly, this region harbors the LKB1 gene, in which germline mutations were shown to be associated with Peutz-Jeghers Syndrome (PJS). PJS and FPH share the disorder of hyperpigmentation, the fine mapping of the FPH gene is expected to lead to a better understanding of the etiology for both FPH and PJS. The linkage of FPH locus to human chromosome 19p13.1-pter provides a genetic basis for further fine mapping. PMID- 16709485 TI - A novel GJA 1 mutation in oculo-dento-digital dysplasia with curly hair and hyperkeratosis. AB - Oculo-dento-digital dysplasia (ODDD) is a rare autosomal dominant congenital disorder mainly affecting the development of the face, eyes, skeletal system, heart and dentition. ODDD has been mapped to chromosome 6q22-q24 and germline mutations have been identified in the connexin 43 gene, GJA1. Abnormalities of the skin, hair, and nails have been recognized in ODDD but are often easily overlooked. We report an ODDD patient with curly hair, early trichorrhexis nodosa and discrete keratoderma. Molecular genetic studies revealed a novel GJA1 mutation affecting the amino terminus of the gap junction protein alpha-1 (Cx43). In the light of the cutaneous findings in our patient and based on recent ectodermal dysplasia classification systems, we propose to include ODDD in the group of ectodermal dysplasias. PMID- 16709487 TI - Higher body mass index is a significant risk factor for acne formation in schoolchildren. AB - Obesity is frequently accompanied by peripheral hyperandrogenism, which may be associated with increased sebum production and the development of severe acne. Body mass index (BMI) is one of the most accurate ways to measure and determine obesity. The aim of the present study was to study the correlation between obesity and the point prevalence and patterns of acne in schoolchildren. A total of 3,274 children (aged 6-11 years) from Magong Township were examined by two board-certified dermatologists. The acne prevalence was 7.3%, with more girls affected than boys (ratio = 1.5). Comedones were more commonly observed than inflammatory acne (10.4% vs. 6.9%). The mean of BMI in non-acne students (18.2 +/ 3.4) was significantly lower than that in acne subjects (19.5 +/- 3.7), without gender difference. Overall schoolchildren with a BMI < 18.5 had less prevalence rate of acne, especially the inflammatory lesions, while those with a BMI-for-age > or = 95% had a significantly higher rate of acne development. PMID- 16709488 TI - Human papillomavirus type 59 identified in a verrucous cyst of the flank. AB - The verrucous cyst is a non-plantar epidermoid cyst with histopathological features of human papillomavirus (HPV) infection, including papillomatosis and hypergranulosis of the cyst lining. We report the first case of a verrucous cyst demonstrating not only the histopathological and immunohistochemical features of HPV infection, but also homology with HPV type 59 on HPV genotyping. A 28-year old male developed a palpable mass in his right flank. Histological examination revealed an intradermal cyst lined by an acanthotic and papillomatous squamous epithelium with prominent keratohyaline granules and squamous eddies. The keratinocyte nuclei were positive for papillomavirus antigens on immunohistochemistry and HPV genotyping demonstrated a homology to HPV type 59, a high-risk genital type. Although we only experienced a single case with such a finding, we suggest that it may be necessary to subject patients with verrucous cysts to a closer follow up for better characterization of their clinical behavior. PMID- 16709489 TI - Unstabilized DNA breaks in lymphocytes of patients with systemic sclerosis. AB - The clastogenic effects on DNA, proven by the presence of micronuclei (MN), and the protective cellular mechanisms normally used to stabilize DNA breaks were investigated in patients with systemic sclerosis (SSc). The frequency of micronucleated cells found in cultures of peripheral lymphocytes in patients was significantly higher than in the control group. The patient group with anti centromere antibodies showed a significantly higher frequency of micronucleated cells than that observed in the patients with anti-topoisomerase I antibodies (4.22% versus 2.34%, p < 0.001). Moreover, we attempted to characterize MN for the presence or absence of DNA fragments with free 3'-OH ends by digoxigenin-dUTP (DIG-dUTP) using terminal deoxynucleotidil transferase. It was found that the frequency of MN containing DNA fragments with 3'-OH free ends (unstable fragments) increased in SSc patients compared to that observed in the control group. Moreover, this increase was significantly higher in lymphocytes of the patients with anti-centromere antibodies than in those with anti-topoisomerase I antibodies (35% versus 20.08%, p < 0.001). Our results indicate that in SSc patients there is an interference in the protective cellular mechanisms, normally stabilizing DNA breaks. PMID- 16709490 TI - Overexpression of phosphorylated-STAT3 and phosphorylated-ERK protein in dermatofibrosarcoma protuberans. AB - The overexpression of phosphorylated signal transducer and activator of transcription-3 (p-STAT3) and phosphorylated extracellular signal-regulated kinase (p-ERK) have recently been shown to play an important role in the pathogenesis of various human tumors. However, the role of these two major signal transduction pathways in dermatofibrosarcoma protuberans (DFSP) remains unknown. This study was designed to investigate the significance of p-STAT3 and p-ERK expression in DFSP. The expressions of p-STAT3 and p-ERK were analyzed by immunohistochemical staining in formalin-fixed, paraffin-embedded tissue sections of human DFSP and dermatofibroma. Ten cases were positive for p-STAT3 expression in 14 cases of DFSP, however, only 5 cases were positive in 20 cases of dermatofibroma. Eleven out of 14 cases of DFSP expressed p-ERK, but only four cases were positive in 20 cases of dermatofibroma. The expressions of p-STAT3 and p-ERK were significantly higher than those in dermatofibroma (both p < 0.01). This study suggests that the overexpression of p-STAT3 and p-ERK may play a pivotal role in the oncogenesis of DFSP. PMID- 16709491 TI - Delayed response of oral pemphigus vulgaris to rituximab treatment. AB - Rituximab is a monoclonal antibody directed against the CD20 antigen of B cells, which has been developed for the treatment of lymphomas and has been successfully used in the treatment of recalcitrant pemphigus vulgaris. Here, we report on a 26 year-old patient with an 18 month history of pemphigus vulgaris of the oral mucosa where treatment with rituximab led to a delayed but sustained therapeutic response. This case is of interest since most of the previous reports have described a more rapid clinical improvement of refractory pemphigus by rituximab treatment. The delayed clinical response to rituximab of the present case may be explained by the persistence of long-lived plasma cells that continued to produce pathogenic autoantibodies against desmoglein 3. PMID- 16709492 TI - Coping with exacerbation in psoriasis and eczema prior to admission in a dermatological ward. AB - Chronic dermatologic diseases, such as psoriasis and eczema, may cause significant psycho-social problems and stress. Our objectives were to characterize how hospitalised patients coped with psoriasis and eczema, and to investigate the relationship between coping and quality of life. Data are based on survey forms completed upon admission to the dermatology ward from 212 patients with chronic dermatological diseases, 146 with psoriasis and 66 with eczema. 108 were men, average age 48 years. The Norwegian versions of the standardized survey questionnaires, Jalowiec Coping Scale and Dermatological Life Quality Index, were used to evaluate coping and quality of life. We found that optimism, belief-in-oneself and confrontational coping strategies were most frequently used. Long duration of the disease was correlated to the belief-in oneself strategy, while short duration was related to supportive strategies. More frequent use of confrontational and optimistic modes was significantly related to better quality of life. More frequent use of emotional and evasive modes was significantly related to poorer quality of life. There was no significant difference between the psoriasis and eczema groups in terms of use of coping strategies, with exception of emotional strategies. Knowledge of coping strategies and quality of life among patients with chronic dermatological diseases is important for improvement in health services for these patients. PMID- 16709493 TI - Spitzoid melanoma in childhood. AB - Spitzoid melanoma is a rare variant of melanoma. It has morphological features similar to those of Spitz's nevus. In this study, the histologic and immunohistochemical features of both Spitzoid melanoma and Spitz's nevus are emphasized. We report two cases of melanoma with spitzoid features occurring in the extremities of 9 and 8-year-old girls. Histologically both lesions had typical features of Spitzoid melanoma. We conclude that the differential diagnosis of Spitzoid melanoma and Spitz's nevus is at times problematic in childhood, in that distant metastasis may be the only diagnostic criteria for some cases to be distinguished from Spitz's nevus if strict criteria are followed. Spitzoid melanoma must be treated as other types of melanoma. PMID- 16709494 TI - Drug-induced cutaneous lupus erythematosus after 5 years of treatment with carbamazepine. AB - Drug-induced lupus is a disease, inducible by several drugs, that shares symptoms and laboratory characteristics with idiopathic lupus erythematosus. We report a case of carbamazepine-induced cutaneous lupus erythematosus which developed after 5 years of treatment in a patient who was on this medication because of an epileptic crisis after cranial trauma. Carbamazepine is a medication rarely implicated in drug-induced cutaneous lupus, moreover there are very few reports of such long periods between the start of therapy and the presentation of the clinical symptoms. We describe this case to emphasize the possibility of cutaneous lupus induction by carbamazepine even after many years of therapy. PMID- 16709495 TI - Two giant orf lesions in a heart/lung transplant patient. AB - Orf is an infectious ulcerative stomatitis of sheep and goats. The responsible pathogen, parapoxvirus, may be transmitted to humans. Orf lesions are often atypical in immunocompromised individuals. The present report describes two very large exophytic lesions in a 31-year-old transplant patient receiving oral tacrolimus, mycophenolate mofetil and prednisone. Early surgical excision was successful, with no relapse after 14 months. PMID- 16709496 TI - Peeling skin syndrome. AB - Peeling skin syndrome is a rare disease characterized by widespread painless peeling of the skin. To date, several cases have been described with different clinical features called peeling skin syndrome. Previous reports describe two types (type A and type B) of peeling skin syndrome, both of which show generalized desquamation, sparing palms and soles. We report a 23-year old man who has been classified as neither type A nor type B, and whose history, clinical features and histopathological findings led to a diagnosis of peeling skin syndrome. In addition, the desquamation pattern in our patient was different from that of both types because our case's palms and soles were involved too. PMID- 16709498 TI - Rosai-Dorfman disease with cutaneous manifestation (sinus histiocytosis with massive lymphadenopathy). AB - We report a 73-year-old woman with the typical findings of "Rosai-Dorfman disease" (RDD) including massive, painless lymphadenopathy, fever and multiple, asymptomatic cutaneous lesions on her face, collum and neck. Histologically, the lesions were marked by characteristic large macrophages exhibiting emperipolesis. PMID- 16709497 TI - Angiosarcoma of the abdominal wall after dermolipectomy in a morbidly obese man. A rare form of presentation of Stewart-Treves syndrome. AB - Angiosarcoma is a rare malignant tumor, with a predilection for skin in the head and neck region, although it has been described in many other locations. Its association with chronic lymphedema is well known, mainly in the setting of postmastectomy lymphedema of the arm in breast cancer patients (termed Stewart Treves syndrome). However, angiosarcoma can appear in lower limbs with chronic lymphedema and rarely in other locations such as the abdominal wall. Herein, we present a unique case of angiosarcoma developing in the abdominal wall of a morbidly obese patient after extensive dermolipectomy. PMID- 16709499 TI - Fusariosis occurring in an ulcerated cutaneous CD8+ T cell lymphoma tumor. AB - Fusarium species have recently emerged as the second most common pathogenic mold in immunocompromised patients, and they are moderately resistant to most antifungal agents. The skin lesions of disseminated fusariosis typically manifest as multiple red or violaceous macules or nodules, often ulcerated and covered by a black eschar. We report a case of cutaneous fusariosis in a patient with long standing hypopigmented mycosis fungoides. The infection was successfully treated with a 3-month course of oral voriconazole. The present case is unusual in that the infection occurred within a pre-existing, ulcerated lesion of cutaneous CD8+ lymphoma, resulting clinically in confusion with pyoderma gangrenosum and necrosis of lymphoma. A high index of suspicion will prompt a timely biopsy as well as isolation of the fungus, and early institution of systemic antifungal therapy. PMID- 16709500 TI - Subacute cutaneous lupus erythematosus on the lines of Blaschko. AB - We describe a 42-year-old woman who had erythemateous plaques with sharply demarcated margins and fine scaling on her right trunk and leg. The lesions had a linear distribution following the lines of Blaschko. Histological findings supported the diagnosis of subacute cutaneous lupus erythematosus (SCLE) on the lines of Blaschko. Treatment with systemic corticosteroids and anti-malarial agents resulted in remission. Up to now, eighteen cases of linear lupus erythematosus have been described. Linear lupus erythematosus mostly affects children presenting with lesions on the scalp that were classified as discoid lupus erythematosus. To our knowledge, this is the first case of subacute cutaneous lupus erythematosus on the lines of Blaschko. PMID- 16709501 TI - Generalized morphea-like lesions arising in mechanically-compressed areas by underclothes. AB - We report two patients with generalized morphea-like eruptions, the distribution of which was confined to areas mechanically compressed by underclothes. One patient had Raynaud's phenomenon and anti-centromere antibodies, and the other patient had a long history of occupational exposure to organic solvents. We believe that the Koebner phenomenon, resulting from local compression by underclothes, might be responsible for the development of morphea-like lesions in patients with subclinical systemic sclerosis. PMID- 16709503 TI - Brachioradial pruritus: report of a new case responding to gabapentin. PMID- 16709502 TI - Cutaneous leishmaniasis in two siblings. PMID- 16709504 TI - Livedo racemosa as a cutaneous manifestation of polycythemia vera. PMID- 16709505 TI - Meibomian gland carcinoma. PMID- 16709506 TI - Penile syringoma in a 14-year-old boy. PMID- 16709507 TI - Excision and split thickness skin grafting in the treatment of nodular primary localized cutaneous amyloidosis. PMID- 16709508 TI - Psoriasis exacerbation after a flu-like syndrome during anti-TNF-alpha therapy. PMID- 16709509 TI - Acute generalized exanthematous pustulosis induced by hydroxychloroquine. PMID- 16709510 TI - Acrodermatitis acidemica with an eating disorder. PMID- 16709511 TI - Retinitis pigmentosa associated with vitiligo. PMID- 16709512 TI - Does cutaneous silica granuloma develop mainly in predisposed patients? PMID- 16709513 TI - Environmental hazards and the skin. PMID- 16709514 TI - Occupational skin diseases. PMID- 16709515 TI - Three cases of renal relapse after allogeneic hematopoietic stem cell transplantation for childhood acute lymphoblastic leukemia. AB - Isolated renal relapse after allogeneic hematopoietic stem cell transplantation (alloHSCT) in children with acute lymphoblastic leukemia (ALL) is a rare condition. Generally, in ALL, the sites most frequently affected by extramedullary relapse are the central nervous system (CNS) and the testicles. Here we report on three young boys with relapsed B-precursor ALL, who underwent alloHSCT from HLA-identical siblings and suffered a histopathologically proven isolated unilateral renal relapse (two patients) or a combined renal and testicular relapse (one patient) 6, 10 and 12 months post alloHSCT. In all patients at the time of relapse bone marrow showed complete remission with complete donor hematopoiesis. They all received total body irradiation with partial shielding of the kidneys as part of their conditioning therapy, such that renal shielding could be an explanation for the observed accumulation of renal relapses. Moreover, during the past few years so called immune privilege has been postulated for frequent relapse sites such as the CNS, the testicles and the anterior chamber of the eye. Impaired accessability of these organs by cytotoxic T-cells (CTLs) with a reduced graft-versus-leukemia (GvL) effect after alloHSCT is based on a number of different molecular and cellular mechanisms. Similar mechanisms have been shown to be effective in the tubulointerstitial space of the kidney, rendering the kidney a potentially immune privileged site. Due to these observations we advocate sufficient treatment of the kidneys during conditioning therapy. PMID- 16709516 TI - Spontaneous splenic rupture in a healthy allogeneic donor of peripheral-blood stem cell following the administration of granulocyte colony-stimulating factor (g-csf). A case report and review of the literature. AB - Human granulocyte colony-stimulating factor (G-CSF) is a hematopoietic hormone promoting the growth, proliferation, differentiation and maturation of myeloid and leukocytic lineages. G-csfs have been used to improve granulocyte count in neutropenic patients, reduce the incidence and duration of neutropenia in patients receiving cytotoxic chemotherapy and to mobilize peripheral blood stem cells prior to leukapheresis for using in both autologous and allogeneic hematopoietic cell transplantation. In general, side-effects are mild to moderate and life threatening side-effects like splenic rupture are very rare. We herein, report a case of spontaneous splenic rupture secondary to high-dose G-CSF use (20 mcg/kg/day), in a healthy female allogeneic donor of peripheral-blood stem cell (PBSC) . PMID- 16709517 TI - Two consecutive immunophenotypic switches in a child with MLL-rearranged acute lymphoblastic leukemia. AB - An 18-month-old girl was diagnosed with pre-pre-B ALL/t(4;11) leukemia, which during the treatment and after matched bone marrow transplantation (BMT), underwent two consecutive switches from lymphoid to myeloid lineage and vice versa. The high expression of HOXA9 and FLT3 genes remaining genotypically stable in a leukemia throughout phenotypic switches, suggests that this leukemia may have originated as a common B/myeloid progenitors. PMID- 16709518 TI - Polyneuropathy, organomegaly, endocrinopathy, M protein, skin changes: not always a POEMS syndrome. PMID- 16709519 TI - Exchange blood transfusions for the treatment of leg ulcerations in thalassemia intermedia. AB - Thalassemia intermedia is a heterogeneous, transfusion-independent form of b thalassemia, with a clinical course dominated by multi-organ effects of chronic tissue hypoxia, in which hemoglobin F percentage seems to play an important role. We describe the case of a transfusion-independent thalassemia intermedia patient (total hemoglobin 10.7 g/dl) with high hemoglobin F percentage (70%), who presented with persistent leg ulcerations. The patient was successfully treated with one-year exchange blood transfusions, which reduced hemoglobin F percentage to 35%. PMID- 16709520 TI - Mixed autoimmune haemolysis in a SLE patient due to aspecific and anti-Jka autoantibodies; case report and review of the literature. PMID- 16709521 TI - Alemtuzumab induced complete remission of autoimmune hemolytic anemia refractory to corticosteroids, splenectomy and rituximab. AB - A 58-year-old man with warm-antibody-mediated autoimmune hemolytic anemia (AIHA) refractory to prednisolone, azathioprine, splenectomy, rituximab and combination chemotherapy, and with unacceptably high transfusion requirement, was treated with alemtuzumab. After a cumulative dose of 883 mg of alemtuzumab, the AIHA remitted completely, with normalization of hemoglobin and transfusion independence. The major side effect was reactivation of cytomegalovirus, which was controlled with intravenous and oral ganciclovir. This case showed that alemtuzumab might be of use in therapy-refractory AIHA. PMID- 16709522 TI - Bone marrow amyloidosis. PMID- 16709525 TI - Non-tuberculous mycobacterial infections. AB - Non-tuberculous mycobacteria (NTM) are ubiquitous environmental organisms in contrast to those belonging to the M. tuberculosi complex (MTB). NTM infects and causes disease only in hosts with local or general predisposing factors. Lung infection following inhalation of NTM is the most common NTM disease but soft tissue infections may occur in connection with contaminated trauma or surgery. Microbiological diagnosis is obtained by microscopy for acid-fast bacteria (AFB) on secretions or biopsies, and by culture on special media. With the high specificity of MTB- polymerase chain reaction (PCR), a positive AFB smear combined with negative MTB-PCR denotes infection with NTM. Sophisticated species diagnosis of cultured NTM is attained by various molecular methods, where 16S rDNA-sequencing remains the gold standard. The panorama of infection with different rapidly growing (RGM) or slowly growing mycobacteria (SGM) in Sweden is described. Sensitivity testing in vitro to antimycobacterial drugs against NTM does not always preclude the in vivo efficacy. Standard antimycobacterial treatment regimens have been defined for infection with several NTM species. Sensitivity testing should be performed in selected cases only, as in case of relapse or suspected development of resistance of the NTM strain. The spectrum of disease caused by NTM species that display a very low pathogenic potential is likely to widen over time as severe immunosuppression will continue to be prevalent in several patient categories. PMID- 16709526 TI - Management of pain in acute otitis media in Finnish primary care. AB - Most treatment recommendations for acute otitis media favour active use of pain relief medication. These data comprised 3059 Finnish primary care acute otitis media patients. We found that 10.4% of the patients were prescribed or recommended analgesics, which is in contrast to treatment recommendations. PMID- 16709527 TI - 'Break-point Checkerboard Plate' for screening of appropriate antibiotic combinations against multidrug-resistant Pseudomonas aeruginosa. AB - Increase of multiple drug resistant Pseudomonas aeruginosa (MDRP) is becoming a serious problem in the clinical setting. Although the checkerboard method to determine FIC index and synergistic effects of antibiotic combinations is useful, it is not well adapted to a routine test, mainly because of its time-consuming and labor-intensive nature. Here we report 'Break-point Checkerboard Plate', in which breakpoint concentrations, such as 'S' (sensitive) and 'I' (intermediate), were combined in a microtiter plate with 8 antibiotics, including carbapenem, aminoglycoside and fluoroquinolone. The results obtained from 12 strains of MDRP demonstrated a strong synergistic effect of some antibiotic combinations at clinically relevant concentrations. Our data suggest a usefulness of 'Break-point Checkerboard Plate' to screen appropriate antibiotic combinations against drug resistant organisms, including MDRP. PMID- 16709528 TI - Antigenic diversity of Neisseria meningitidis isolated in Taiwan between 1995 and 2002. AB - The geographic distribution of N. meningitidis is diverse. Information on the antigenic variation of N. meningitidis is important for the development of an outer membrane protein-based vaccine. As a first step towards vaccine development, serological typing was performed to determine the antigenic properties of 127 invasive N. meningitidis isolates collected in Taiwan between 1995 and 2002. With 31.5% non-serotypeable and 32.3% non-serosubtypeable, the 127 isolates fell into 51 phenotypes, with W135:NT:P1.5,2:L3,7,9, Y:14P1.5,2, and B:1:NST:L3,7,9 being the 3 most prevalent. Among the 37 serogroup B isolates, 15 serosubtypes were found, with P1.5,2 and P1.12,13 being the most prevalent. The high diversity of Por A among serogroup B isolates circulating in Taiwan poses a great challenge for the development of a PorA-based vaccine. Because 85% of the serogroup B isolates had the L3,7,9 immunotype, inclusion of L3,7,9 lipooligosaccharides in a PorA-based vaccine may be a promising approach. In addition, based on the phenotypic characterization, we suggest that both serogroup B and W135 isolates were endemic and that serogroup A, C, and Y isolates were imported, which may reflect increased international travel. PMID- 16709529 TI - Daptomycin for the treatment of vancomycin resistant Enterococcus faecium bacteremia. AB - The best therapeutic options for serious infections due to vancomyci resistant Enterococcus (VRE) remain unclear. We describe the successful treatment of vancomycin resistant Enterococcus faecium bacteremia in 2 patients using daptomycin. We also briefly review the literature on antibiotic options for VRE infection including the use of daptomycin. PMID- 16709530 TI - Lemierre's syndrome: still an important clinical entity. AB - We report a case of Lemierre's syndrome caused by Fusobacterium necrophorum and discuss characteristics of this potentially fatal condition which, though rare, may have a rising frequency. Familiarity with the signs and symptoms of Lemierre's syndrome is therefore of great importance. PMID- 16709531 TI - Isolation of Cardiobacterium hominis from the peritoneal fluid of a patient on continuous ambulatory peritoneal dialysis. AB - Cardiobacterium hominis, an uncommon cause of bacterial endocarditis, is rarely implicated in infections outside the vascular system. We report the isolation of C. hominis from the peritoneal fluid of a patient on continuous ambulatory peritoneal dialysis with a presentation suggestive of peritonitis but no evidence of infective endocarditis. PMID- 16709532 TI - Disseminated cytomegalovirus infection associated with Pneumocystis carinii pneumonia in a previously normal infant. AB - We reported a rare case of dual opportunistic infections of Pneumocystis carinii pneumonia and disseminated cytomegalovirus (CMV) infection in a previously normal female infant. Transient T-cell dysfunction was demonstrated and returned to normal after treatment of CMV infection. PMID- 16709533 TI - Probiotic lactobacilli and bacteraemia in Stockholm. AB - The objective of the present study was to determine the incidence of lactobacilli induced bacteraemia in Stockholm, Sweden, between January 1998 and March 2004, and to identify the possible presence of probiotic strains. Isolated strains were checked for growth on selective lactobacilli-agar and the biochemical profiles were determined. The lactobacilli strains were further analysed with molecular methodologies to ascertain eventual similarities with the probiotic strains Lactobacillus paracasei subsp. paracasei F19 (LMG P 17806), Lactobacillus acidophilus NCFB 1748 and Lactobacillus rhamnosus GG (ATCC 53103). The minimum inhibitory concentrations of selected antimicrobial agents were determined by the agar dilution method. The incidence of bacteraemia cases caused by lactobacilli remained at the same level during the study period and constituted to <1% of the total number of bacteraemia cases each y. L. paracasei subsp. paracasei F19, L. acidophilus NCFB 1748 and L. rhamnosus GG were not identified in any of the samples. PMID- 16709534 TI - Genotypic analysis of Acinetobacter bloodstream infection isolates in a Turkish university hospital. AB - Acinetobacter baumannii is a significant pathogen of bloodstream infections in hospital patients that frequently causes single clone outbreaks. We aimed to evaluate the genetic relatedness and antimicrobial susceptibility of Acinetobacter spp. bloodstream isolates, in order to obtain insight into their cross-transmission. This prospective study was conducted at the Erciyes University Hospital. During a 1-y period, all patients with nosocomial BSI caused by Acinetobacter spp. were included in the study. All data with regard to the patients, underlying diseases and risk factors for BSI and the severity of disease were collected. Blood culture isolates of Acinetobacter spp. were identified according to their morphology and biochemical reactions. The antimicrobial susceptibility was determined using the Kirby-Bauer disk diffusion test according to the NCCLS; the genetic relatedness of isolates was determined by RAPD-PCR analysis and pulsed-field gel electrophoresis (PFGE). 41 patients acquired a nosocomial bloodstream infection caused by A. baumanii during this period. 88% of these infections (36 of 41) occurred while the patients were treated in the intensive care unit. Nearly 80% of the isolates belonged to 3 genotypes, suggesting cross-transmission in ICU settings where infection control practices are poor. All Acinetobacter isolates were multidrug-resistant and the crude mortality of patients infected with A. baumanii was 80.5%. We concluded that the genetic relatedness of Acinetobacter spp. causing BSI was very high, indicating cross-transmission within the ICU setting. Essential components of an infection control programme to prevent nosocomial transmission of A. baumannii are early detection of colonized patients, followed by strict attention to standard precautions and contact isolation. PMID- 16709535 TI - Suction drainage culture as a guide to effectively treat musculoskeletal infection. AB - Following surgery for musculoskeletal infection, a positive suction drainage culture (SDC) is consistent with persistent sepsis. Our objective was to determine the effect of a negative SDC obtained in subsequent operations on the outcome of a musculoskeletal infection. 99 patients were prospectively enrolled, all treated surgically for musculoskeletal infection utilizing suction drainage and appropriate antimicrobial therapy. Surgery consisted of irrigation, debridement, and prosthetic exchange or implant removal. SDC was considered negative if all bottles resulted in negative cultures. Following SDC results, patients were placed into 1 of 2 treatment groups: 1) Negative SDC, and no new operation; or 2) Positive SDC, and new operation(s) until SDC was negative. Antibiotic therapy ranged from 6-12 weeks (osteomyelitis) to 10-21 d (soft tissue). Both groups were similar with regard to baseline characteristics. Cure was obtained in 91.8% of patients (56/61) in group 1 and 91.6% of patients (22/24) in group 2. Similar results were obtained in patients with an infection in the presence of an implant. In conclusion, a negative SDC following surgery for a musculoskeletal infection is a strong indication of eventual outcome. PMID- 16709536 TI - Eosinophil cationic protein in patients with fascioliasis: its probable effects on symptoms and signs. AB - Eosinophilia is 1 of the most common signs of fascioliasis especially during acute stage. In this study, our aim was to determine eosinophil cationic protein (ECP) in the sera of patients with fascioliasis and to investigate the likely relation between ECP and symptoms and signs of fascioliasis. Presence of high liver enzyme levels, eosinophilia, abdominal pain or urticaria was not found to be significantly associated with ECP positivity (p>0.05), but statistical analyses revealed that ECP positivity was significantly related to weight loss status of patients (p<0.005). In conclusion, ECP may be 1 of the likely causes of the symptoms and signs of fascioliasis. PMID- 16709537 TI - Safety and immunogenicity of a primary course and booster dose of a combined diphtheria, tetanus, acellular pertussis, hepatitis B and inactivated poliovirus vaccine. AB - Primary immunization at 3, 4.5, and 6 months and boosting between 15 and 27 months of age with combined diphtheria-tetanus-acellular pertussis-hepatitis B inactivated poliovirus (DTPa-HBV-IPV) vaccine was compared with separate administration of DTPa-HBV and IPV to healthy children (trials DTPa-HBV-IPV 019/033). Antibody titres were measured before and 1 month after primary and booster courses. Solicited local and general symptoms were recorded using diary cards. One month after primary vaccination, all children in both groups developed antibody titres above the assay cut-off for all vaccine components. Significantly higher anti-diphtheria, anti-pertactin (PRN) and anti-polio GMTs were measured following DTPa-HBV-IPV than DTPa-HBV plus IPV. Prior to boosting similar seroprotection/seropositivity rates were recorded in both groups. After boosting all children had seroprotective levels of diphtheria, tetanus, polio and HBV. Criteria for pertussis vaccine response were fulfilled in most children. Significantly higher anti-PRN GMTs were measured following DTPa-HBV-IPV than DTPa HBV plus IPV. There was no difference between groups in the incidence or intensity of local and general symptoms after primary or booster vaccination, except for fever which was more frequent after the booster dose in the combined vaccine group. Both vaccine regimens were well tolerated and immunogenic, however the combined administration has the advantage of being administered as a single injection. PMID- 16709538 TI - Minimal change in HSV-2 seroreactivity: a cross-sectional Swedish population study. AB - The prevalence of antibodies to herpes simplex virus type 2 (HSV-2) was determined in sera collected from 2 Swedish cross-sectional populations during 1990/91 and 1996/97. A glycoprotein G2 (gG-2) peptide ELISA was used to analyse 2899 sera from adults. We found a prevalence of 13.0% in 18-70-y-olds in 1990/91 and 16.4% in 31-55-y-olds in 1996/97. Seropositivity to HSV-2 was somewhat higher in women than in men; 14.4% compared to 11.6% in 1990/91 and 18.9% compared to 13.6% in 1996/97. Overall there was a significant difference in female prevalence of HSV-2 compared to prevalence in males of the same age groups (p=0.013). Total antibody prevalence to HSV-1 of 88% in HSV-2 negative individuals above 18 y of age was found. HSV-2 positive individuals were found seropositive to HSV-1 in approximately 95% of the cases. 12% were negative for both HSV-1 and HSV-2 antibodies. In addition, there was no serological sign that the reported increase of HSV-1 as an early sexual disease had reduced the frequency of HSV-2 seropositivity in the sexually most active age groups. In conclusion, our investigation has shown no significant increase of HSV-2 seroprevalence in randomly selected Swedish subjects during the defined time period. PMID- 16709539 TI - Current status of inpatient HIV care at a tertiary care center in Nepal. AB - The success of the WHO's '3 by 5' programme will depend on the ability of developing countries to provide antiretroviral therapy. The purpose of this study was to determine the current status of HIV care at a major teaching hospital in eastern Nepal. A retrospective cohort study was conducted of admitted HIV infected patients at BPKIHS between 1993 and 2003, evaluating reasons for admission and the care received. There were 2, 2, 2, 3, 5 and 18 admissions, respectively, from 1998 to 2003. Only 4 were female patients. Two were children, both admitted in the last y studied. 10 admissions (31% of the total) were for opportunistic infections (OIs). Seven patients were prescribed OI prophylaxis, with incorrect dosages in 6. Four patients were prescribed antiretroviral therapy, all in inappropriate dosages or combinations. 13 patients were rapidly discharged without further care as soon as a diagnosis of HIV was made. Hospital admissions of HIV-infected individuals are increasing rapidly. Important segments of the HIV-infected population such as women and children are not receiving medical services. Most admissions are not due to OIs. The care of HIV-infected patients even at a major tertiary care teaching hospital in Nepal is sub-optimal. PMID- 16709540 TI - Efficacy of nelfinavir-based treatment in the central nervous system of HIV-1 infected patients. AB - We studied the HIV-1 load and nelfinavir (NFV) concentrations in cerebrospinal fluid (CSF) after long-term successful NFV-based therapy, using ultrasensitive methods of detection. 19 patients without virological failure in plasma, who had been treated with 2 nucleoside analogue reverse transcripaste inhibitors (NRTI) and NFV for a minimum of 18 months were included. HIV-RNA was determined in plasma and CSF using an ultrasensitive method (<2 copies/ml). Total and free concentrations of NFV were analysed using high liquid chromatography with UV light detection. 12 out of 19 (63%) patients had <2 copies HIV-RNA/ml in CSF. Seven subjects ranged between 3 and 39 copies/ml, 2 of whom had a slightly higher viral load in CSF than in plasma. NFV was detected in CSF in 16 out of 18 patients analysed and was quantifiable in 8 patients, at concentrations ranging from 6 to 29 nM. There was no correlation between NFV concentration and HIV-RNA levels. Long-term therapy with NFV + 2 NRTI showed no increased rate of virological treatment failure within the central nervous system (CNS) in compliant patients, despite earlier reports of lack of NFV penetration to CNS. Using a highly sensitive method, NFV was detected and quantified in the CSF, although at low values, which could have contributed to the high anti-HIV-1 efficacy of the therapy seen in our subjects. PMID- 16709541 TI - Meningitis due to Capnocytophaga canimorsus: contribution of 16S RNA ribosomal sequencing for species identification. AB - We report a case of Capnocytophaga canimorsus meningitis, which is distinguishable from previous reports by the use of 16S rRNA sequencing for species identification. Our case report highlights the interest of molecular new tools for bacteriological diagnosis of human infections. PMID- 16709542 TI - Actinomycosis or tuberculosis? A diagnostic dilemma. AB - Actinomycosis of spine is a very rare disease. Very few cases have been studied and reported in the past. The dilemma of distinguishing the condition from other disorders relies on the competency of the treating physician and a proper knowledge of the subtle radiological differences between these disorders especially in underdeveloped and developing countries where tuberculosis still has a very strong foothold. A rare atypical case of actinomycosis of spine resembling tuberculosis is presented. PMID- 16709543 TI - Effectiveness of linezolid and imipenem association in the treatment of severe community-acquired pneumonia in children: two case reports. AB - Two children with severe pneumonia, purulent pleural effusions, and abscess formation unresponsive to appropriate antibiotic therapy recovered promptly after the introduction of linezolid and imipenem association. Linezolid is a new antibiotic with high bioavailability and an outstanding safety profile, synergistic with imipenem, which may deserve a place in the armamentarium for severe pneumonia in children. PMID- 16709544 TI - First report of a linezolid- and vancomycin-resistant Enterococcus faecium strain in Korea. AB - We report the first clinical isolate of linezolid- and vancomycin-resistant Enterococcus faecium (VREF) in Korea. The minimum inhibitory concentration of linezolid was in direct proportion to the number of copies of the G2576U mutation according to sequencing and restriction fragment length polymorphism analysis of the 23S rRNA gene. PMID- 16709545 TI - Paragonimiasis presenting with unilateral pseudochylothorax: case report and literature review. AB - A 64-y-old female previously treated for lung fluke presented with shortness of breath and moderate left pleural effusion. Thoracentesis was performed and milky pleural effusion was obtained. Paragonimus ova and rhomboidal cholesterol crystals were found. Paragonimiasis was diagnosed, and the patient was treated with praziquantel 25 mg/kg/d. PMID- 16709546 TI - Long-term home-based parenteral antibiotic treatment of a prosthetic vascular graft infection caused by Pseudomonas aeruginosa. AB - Survival for more than 5 y was obtained with home-based parenteral antibiotic therapy in a 68-y-old male with Pseudomonas aeruginosa infection of an aortic bifemoral prosthetic graft. Surgical intervention was deemed to carry to high a risk and we suggest that lifelong suppressive antibiotic therapy may be an option in selected patients. PMID- 16709547 TI - Human metapneumovirus as a causative agent of lower respiratory tract infection in four patients: the first report of human metapneumovirus infection confirmed by RNA sequences in Taiwan. AB - Human metapneumovirus (HMPV) has been described as a human pathogen associated with respiratory disease in most parts of the world from all age groups. This report presents the first 4 cases of HMPV disease confirmed by the sequences of PCR products in Taiwan. PMID- 16709548 TI - Primary meningococcal arthritis in a child: case report and literature review. AB - We present a case of primary meningococcal arthritis in an 8-month-old immunocompetent female. We also review 18 additional paediatric cases and characterize this unique form of meningococcal disease. PMID- 16709550 TI - The Female Sexual Function Index (FSFI) and the Female Sexual Distress Scale (FSDS): psychometric properties within a Dutch population. AB - The aim of the present study was to investigate the psychometric properties of the Female Sexual Function Index (FSFI; Rosen et al., 2000) and the Female Sexual Distress Scale (FSDS; Derogatis, Rosen, Leiblum, Burnett, & Heiman, 2002) within a Dutch population of approximately 350 women with and without sexual complaints. The main conclusions of this study are that the multidimensional structure of the FSFI and the unidimensional structure of the FSDS are fairly well replicated in a Dutch sample. The amount of variance explained by confirmatory and exploratory factor analyses was good. Internal consistency and stability of the FSFI and its subscales and the FSDS are satisfactory to good, and the subscales are reasonably stable across demographic variables. The discriminant validity and the ability of the scales to predict the presence or absence of sexual complaints was excellent. The convergent and divergent construct validity of the FSFI and the FSDS was good. These results support the reliability and psychometric validity of the FSFI and the FSDS in the assessment of dimensions of female sexual functioning and sexual distress in clinical and nonclinical samples. PMID- 16709549 TI - Convergent and discriminant validity of clitoral color Doppler ultrasonography as a measure of female sexual arousal. AB - We examined the convergent and discriminant validity of clitoral ultrasonography as a measure of female sexual arousal by assessing the following: (a) its ability to discriminate between sexual and other forms of arousal; and (b) its correlation with subjective ratings of arousal. Results from 63 healthy premenopausal women indicate that ultrasonography was not successful in differentiating sexual arousal from a humor control condition. Furthermore, there were no significant correlations between clitoral blood-flow measures and subjective sexual arousal. Additional research is required to establish the specificity of ultrasonography as a measure of sexual arousal. PMID- 16709551 TI - Hormonal contraception and sexual desire: a critical review. AB - This paper reviews the medical literature from the past thirty years to examine the relationship between hormonal contraception and sexual desire. Although sexual side effects have been noted in various subgroups of women using hormonal contraception, no consistent pattern of effect exists to suggest a hormonal or biological determinant. Effects on sexual desire most likely represent a complex and idiosyncratic combination of biological, psychological, and social effects. Further research is required to identify which factors may have the greatest effect. PMID- 16709553 TI - Genital anaesthesia persisting six years after sertraline discontinuation. AB - Sexual side-effects, in general, are common with selective serotonin-reuptake inhibitors (SSRIs). Genital anaesthesia is a rare side-effect previously described with sertraline and fluoxetine use. With SSRI discontinuation, the sexual side-effects are expected to resolve. We report a case of a 26-year-old male who experienced genital anaesthesia during sertraline treatment and continued to be symptomatic despite medication discontinuation 6 years previously. To date, there have been no published reports of SSRI-induced sexual side-effects persisting beyond SSRI discontinuation. This case highlights the complex interplay of psychopharmacologic and psychodynamic factors that can occur in patients with sexual dysfunction. PMID- 16709552 TI - A randomized open-label comparison of the impact of olanzapine versus risperidone on sexual functioning. AB - The objective of this study was to compare sexual functioning in patients treated with olanzapine or risperidone. This open-label trial included 46 patients randomized to olanzapine (5-15 mg/d) or risperidone (1-6 mg/d) for 6 weeks. We used sexual dysfunction was assessed by a semistructured interview based on the items of the UKU side effect rating scale. Three olanzapine-treated patients (12.0%), compared with 11 risperidone-treated patients (52.4%), reported sexual dysfunctions (p = .008) in the semistructured interview. Only 4 patients (8.7%) spontaneously reported sexual dysfunction. The mean dose was 9.4 mg/d for olanzapine and 3.4 mg/d for risperidone. The mean (+/-SD) prolactin levels (ng/mL) in olanzapine-and risperidone-treated patients were 25.1 (+/- 23.5) and 43.5 (+/- 26.1), respectively. Less sexual dysfunction occurred in the group treated with olanzapine compared with the risperidone group. Direct questioning about sexual functioning is necessary to avoid underestimating the frequency of sexual side effects in patients with schizophrenia and related psychotic disorders. PMID- 16709554 TI - Sexual activity, sexual disorders and associated help-seeking behavior among mature adults in five Anglophone countries from the Global Survey of Sexual Attitudes and Behaviors (GSSAB). AB - We conducted a telephone survey during 2001-2002 to investigate sexual behavior, sexual dysfunction, and related help-seeking behavior in five Anglophone countries. In total, 5,998 individuals (2,992 men and 3,006 women) aged 40-80 years in the United States, Canada, the United Kingdom, Australia, and New Zealand completed the survey. The majority of subjects were sexually active, and although many men and women had experienced sexual dysfunctions, few had sought medical care for these dysfunctions. Further investigation is required to determine whether national differences in the prevalence of sexual dysfunctions are due to genetic predisposition, health factors, cultural perception or socioeconomic situation. PMID- 16709558 TI - One-to-one and polyrhythmic temporal coordination in bimanual circle tracing. AB - The authors manipulated movement amplitude in a bimanual circle-tracing task to alter the natural tracing frequency of the arms. Participants (N = 14) traced different-diameter circles simultaneously with the two arms in either in-phase (0 degrees) or antiphase (180 degrees) coordination, using the index fingers or plastic styli. Movement amplitude altered the natural tracing frequency of the arms, as demonstrated by the following 2 findings: (a) The larger the difference in circle diameter, the larger was the shift from the fixed-point values of 0 degrees and 180 degrees, and the shift increased as movement frequency increased. Those results are consistent with the manipulation of delta omega in the bimanual pendulum paradigm. (b) Increasing movement frequency induced transitions from 1:1 to non-1:1 coordination, contrary to findings in previous investigations of polyrhythmic coordination. Tactile feedback played a minimal role in stabilizing bimanual coordination in the current tasks. PMID- 16709559 TI - Observational modeling effects for movement dynamics and movement outcome measures across differing task constraints: a meta-analysis. AB - The authors conducted a meta-analysis of the observational modeling literature to quantify overall between-participants treatment effects obtained when movement behaviors are acquired. Effects were obtained and reported separately for movement dynamics (MD) and movement outcome (MO) measures. The overall mean observational modeling treatment effects (delta(u)Bi) were 0.77 and 0.17, respectively, for MD and MO measures. The effects reflected a significant advantage of observational modeling over practice-only control conditions. Most important, the magnitude of the obtained effects was far stronger for MD than for MO measures, confirming a distinctive response to observational modeling during motor learning. The advantage for MD measures over observational modeling measures was replicated for different types of tasks. Observational modeling was particularly beneficial for serial tasks (delta(u)Bi = 1.62 and 0.61, respectively, for MD and MO). There were slightly reduced effects for continuous tasks (delta(u)Bi = 1.01 and 0.51, respectively, for MD and MO) and smaller to medium-sized effects for discrete tasks (delta(u)Bi = 0.56 and 0.10, respectively, for MD and MO). The authors discuss those findings with reference to the visual perception perspective on observational modeling, i.e., that demonstrations primarily convey relative motions required to approximate modeled movement behaviors. PMID- 16709560 TI - Discovery of the pendulum and spring dynamics in the early stages of walking. AB - The authors investigated the self-selected, overground walking patterns of 7 children (aged 11 months to 1 year, 5 months) at the initiation of walking (brand new walkers [BNWs]) and for the next 6 months at 1-month intervals. Walking speed, stride length, and stride frequency increased significantly between the first 2 visits without significant changes in height and weight. The authors calculated sagittal plane angular accelerations of the center of mass over the foot for each step as an indicator of the escapement pulse. Results for the acceleration profiles changed after the 1st visit to positive, single-peaked accelerations that occurred < 0.20 s after initial foot contact. Increases in sagittal plane hip angular displacement and decreases in frontal plane pelvic angular displacement were observed. The pattern changes suggest that children quickly discover appropriately timed and directed escapements that initiate and support the conservative sagittal plane pendulum and spring dynamics observed in older children. PMID- 16709561 TI - Influence of endogenous and exogenous orientations of attention on inhibition of return in a cross-modal target-target aiming task. AB - The authors conducted 2 experiments in which participants (N = 16 in each) executed successive unimanual aiming movements to target locations that were indicated by the onset of either an auditory or a visual stimulus. In Experiment 1 (exogenous orientation), inhibition of return (IOR) effects were observed, with reliable reaction time (RT) costs associated with movements returning to the same target and a trend toward larger IOR effects in left than in right space. There was no influence of stimulus modality on the magnitude of IOR. IOR was also observed in Experiment 2 (endogenous orientation), except the influence of stimulus modality reliably mediated those effect. In that case, IOR was evident only when the previous modality was visual and the current modality was auditory. Together, the results of those 2 experiments suggest that in situations in which 2 paired movements constitute the response criteria, IOR is both supramodal and lateralized to contralateral space. PMID- 16709562 TI - Early stage of walking: development of control in mediolateral and anteroposterior directions. AB - The authors examined the changes in bipedal gait of toddlers in the anteroposterior (AP) and mediolateral (ML) directions, as a set, at the onset of independent gait and 1 month after onset. Two groups with distinctly different dynamic resources were studied: 8 toddlers with typical development (TD) and 8 toddlers with Down syndrome (DS). Three-dimensional kinematic data were collected, and gait parameters, such as walking speed, stride length, and stride frequency, as well as the ratio of exchange between potential energy and kinetic energy of the center of mass (COM), were calculated. Displacement of the COM in the AP and ML directions were also analyzed. For some gait variables, toddlers with DS seemed to show more mature values at walking onset than their peers with TD. Those group differences reversed and increased by Visit 2. When the authors considered the motion of the COM of the system, it became clear that the qualitative differences between those groups were characterized primarily by constraints in the ML direction. The authors propose that establishment of coupling between AP and ML oscillations is a key component for the emergence of independent bipedal walking for both populations. PMID- 16709563 TI - Oral kinesthetic deficit in adults who stutter: a target-accuracy study. AB - The current study was based on the hypothesis that chronic developmental stuttering in adults involves a deficiency in oral kinesthesia. The authors used a target-accuracy task to compare oral kinesthesia in adults who stutter (n = 17) and in normal speakers (n = 17). During the task, participants were instructed to make accurate jaw-opening movements in visual and nonvisual feedback conditions. The authors further contrasted oral movement control in a normal response time condition with that in a reaction time condition. Overall, the adults who stutter consistently made significantly less accurate and more variable movements than the control participants in the nonvisual condition, but particularly in the reaction time condition. In general, the present findings suggest that chronic developmental stuttering involves an oral kinesthetic deficiency, although without direct measures of somatosensory function, one cannot exclude a motor deficit interpretation. PMID- 16709564 TI - Identification and validation of mannose 6-phosphate glycoproteins in human plasma reveal a wide range of lysosomal and non-lysosomal proteins. AB - Acid hydrolase activities are normally confined within the cell to the lysosome, a membrane-delimited cytoplasmic organelle primarily responsible for the degradation of macromolecules. However, lysosomal proteins are also present in human plasma, and a proportion of these retain mannose 6-phosphate (Man-6-P), a modification on N-linked glycans that is recognized by Man-6-P receptors (MPRs) that normally direct the targeting of these proteins to the lysosome. In this study, we purified the Man-6-P glycoforms of proteins from human plasma by affinity chromatography on immobilized MPRs and characterized this subproteome by two-dimensional gel electrophoresis and by tandem mass spectrometry. As expected, we identified many known and potential candidate lysosomal proteins. In addition, we also identified a number of abundant classical plasma proteins that were retained even after two consecutive rounds of affinity purification. Given their abundance in plasma, we initially considered these proteins to be likely contaminants, but a mass spectrometric study of Man-6-phosphorylation sites using MPR-purified glycopeptides revealed that some proportion of these classical plasma proteins contained the Man-6-P modification. We propose that these glycoproteins are phosphorylated at low levels by the lysosomal enzyme phosphotransferase, but their high abundance results in detection of Man-6-P glycoforms in plasma. These results may provide useful insights into the molecular processes underlying Man-6-phosphorylation and highlight circumstances under which the presence of Man-6-P may not be indicative of lysosomal function. In addition, characterization of the plasma Man-6-P glycoproteome should facilitate development of mass spectrometry-based tools for the diagnosis of lysosomal storage diseases and for investigating the involvement of Man-6-P containing glycoproteins in more widespread human diseases and their potential utility as biomarkers. PMID- 16709565 TI - The Brichos domain-containing C-terminal part of pro-surfactant protein C binds to an unfolded poly-val transmembrane segment. AB - Native lung surfactant protein C (SP-C) is a 4.2-kDa acylpeptide that associates with alveolar surfactant phospholipids via a transmembrane alpha-helix. This helix contains mainly Val, although poly-Val is inefficient in helix formation, and helical SP-C can spontaneously convert to beta-sheet aggregates and amyloid like fibrils. SP-C is cleaved out from a 21-kDa integral membrane protein, proSP C, in the alveolar type II cell. Recently several mutations localized in the endoplasmic reticulum-lumenal (C-terminal) part of proSP-C (CTproSP-C) have been associated with intracellular accumulation of toxic forms of proSP-C, low levels of mature SP-C, and development of interstitial lung disease. CTproSP-C contains a approximately 100-residue Brichos domain of unknown function that is also found in other membrane proteins associated with amyloid formation, dementia, and cancer. Here we find that recombinant CTproSP-C binds lipid-associated SP-C, which is in beta-strand conformation, and that this interaction results in an increased helical content. In contrast, CTproSP-C does not bind alpha-helical SP C. Recombinant CTproSP-C(L188Q), a mutation associated with interstitial lung disease, shows secondary and quaternary structures similar to those of wild type CTproSP-C but is unable to bind lipid-associated beta-strand SP-C. Transfection of CTproSP-C into HEK293 cells that express proSP-C(L188Q) increases the amount of proSP-C protein, whereas no effect is seen on cells expressing wild type proSP C. These findings suggest that CTproSP-C binds nonhelical SP-C and thereby prevents beta-sheet aggregation and that mutations in CTproSP-C can interfere with this function. PMID- 16709566 TI - Mathematical modeling of polyamine metabolism in mammals. AB - Polyamines are considered as essential compounds in living cells, since they are involved in cell proliferation, transcription, and translation processes. Furthermore, polyamine homeostasis is necessary to cell survival, and its deregulation is involved in relevant processes, such as cancer and neurodegenerative disorders. Great efforts have been made to elucidate the nature of polyamine homeostasis, giving rise to relevant information concerning the behavior of the different components of polyamine metabolism, and a great amount of information has been generated. However, a complex regulation at transcriptional, translational, and metabolic levels as well as the strong relationship between polyamines and essential cell processes make it difficult to discriminate the role of polyamine regulation itself from the whole cell response when an experimental approach is given in vivo. To overcome this limitation, a bottom-up approach to model mathematically metabolic pathways could allow us to elucidate the systemic behavior from individual kinetic and molecular properties. In this paper, we propose a mathematical model of polyamine metabolism from kinetic constants and both metabolite and enzyme levels extracted from bibliographic sources. This model captures the tendencies observed in transgenic mice for the so-called key enzymes of polyamine metabolism, ornithine decarboxylase, S-adenosylmethionine decarboxylase and spermine spermidine N acetyl transferase. Furthermore, the model shows a relevant role of S adenosylmethionine and acetyl-CoA availability in polyamine homeostasis, which are not usually considered in systemic experimental studies. PMID- 16709567 TI - Staphylococcus aureus protein A activates TNFR1 signaling through conserved IgG binding domains. AB - Staphylococcus aureus continues to be a major cause of infection in normal as well as immunocompromised hosts, and the increasing prevalence of highly virulent community-acquired methicillin-resistant strains is a public health concern. A highly expressed surface component of S. aureus, protein A (SpA), contributes to its success as a pathogen by both activating inflammation and by interfering with immune clearance. SpA is known to bind to IgG Fc, which impedes phagocytosis. SpA is also a potent activator of tumor necrosis factor alpha (TNF-alpha) receptor 1 (TNFR1) signaling, inducing both chemokine expression and TNF-converting enzyme dependent soluble TNFR1 (sTNFR1) shedding, which has anti-inflammatory consequences, particularly in the lung. Using a collection of glutathione S transferase fusions to the intact IgG binding region of SpA and to each of the individual binding domains, we found that the SpA IgG binding domains also mediate binding to human airway cells. TNFR1-dependent CXCL8 production could be elicited by any one of the individual SpA IgG binding domains as efficiently as by either the entire SpA or the intact IgG binding region. SpA induction of sTNFR1 shedding required the entire IgG binding region and tolerated fewer substitutions in residues known to interact with IgG. Each of the repeated domains of the IgG binding domain can affect multiple immune responses independently, activating inflammation through TNFR1 and thwarting opsonization by trapping IgG Fc domains, while the intact IgG binding region can limit further signaling through sTNFR1 shedding. PMID- 16709568 TI - Transition from dimers to higher oligomeric forms occurs during the ATPase cycle of the ABCA1 transporter. AB - Fluorescence resonance energy transfer and native PAGE analytical techniques were employed to assess the quaternary structure of ABCA1, an ATP binding cassette transporter playing a crucial role in cellular lipid handling. These experimental approaches support the conclusion that ABCA1 is associated in dimeric structures that undergo transition into higher order structures, i.e. tetramers, during the ATP catalytic cycle. Our data hence underline molecular assembly as a crucial parameter in ABCA1 function and the advantage of native PAGE as analytical tool for intractable membrane proteins. PMID- 16709569 TI - Protective signaling by activated protein C is mechanistically linked to protein C activation on endothelial cells. AB - Activated protein C (APC) has endothelial barrier protective effects that require binding to endothelial protein C receptor (EPCR) and cleavage of protease activated receptor-1 (PAR1) and that may play a role in the anti-inflammatory action of APC. In this study we investigated whether protein C (PC) activation by thrombin on the endothelial cell surface may be linked to efficient protective signaling. To minimize direct thrombin effects on endothelial permeability we used the anticoagulant double mutant thrombin W215A/E217A (WE). Activation of PC by WE on the endothelial cell surface generated APC with high barrier protective activity. Comparable barrier protective effects by exogenous APC required a 4 fold higher concentration of APC. To demonstrate conclusively that protective effects in the presence of WE are mediated by APC generation and not direct signaling by WE, we used a PC variant with a substitution of the active site serine with alanine (PC S360A). Barrier protective effects of a low concentration of exogenous APC were blocked by both wildtype PC and PC S360A, consistent with their expected role as competitive inhibitors for APC binding to EPCR. WE induced protective signaling only in the presence of wild type PC but not PC S360A and PAR1 cleavage was required for these protective effects. These data demonstrate that the endogenous PC activation pathway on the endothelial cell surface is mechanistically linked to PAR1-dependent autocrine barrier protective signaling by the generated APC. WE may have powerful protective effects in systemic inflammation through signaling by the endogenously generated APC. PMID- 16709570 TI - Induction and function of the phage shock protein extracytoplasmic stress response in Escherichia coli. AB - The phage shock protein (Psp) F regulon response in Escherichia coli is thought to be induced by impaired inner membrane integrity and an associated decrease in proton motive force (pmf). Mechanisms by which the Psp system detects the stress signal and responds have so far remained undetermined. Here we demonstrate that PspA and PspG directly confront a variety of inducing stimuli by switching the cell to anaerobic respiration and fermentation and by down-regulating motility, thereby subtly adjusting and maintaining energy usage and pmf. Additionally, PspG controls iron usage. We show that the Psp-inducing protein IV secretin stress, in the absence of Psp proteins, decreases the pmf in an ArcB-dependent manner and that ArcB is required for amplifying and transducing the stress signal to the PspF regulon. The requirement of the ArcB signal transduction protein for induction of psp provides clear evidence for a direct link between the physiological redox state of the cell, the electron transport chain, and induction of the Psp response. Under normal growth conditions PspA and PspD control the level of activity of ArcB/ArcA system that senses the redox/metabolic state of the cell, whereas under stress conditions PspA, PspD, and PspG deliver their effector functions at least in part by activating ArcB/ArcA through positive feedback. PMID- 16709571 TI - K+-independent actions of diazoxide question the role of inner membrane KATP channels in mitochondrial cytoprotective signaling. AB - Activation by diazoxide and inhibition by 5-hydroxydecanoate are the hallmarks of mitochondrial ATP-sensitive K+ (K(ATP)) channels. Opening of these channels is thought to trigger cytoprotection (preconditioning) through the generation of reactive oxygen species. However, we found that diazoxide-induced oxidation of the widely used reactive oxygen species indicator 2',7' dichlorodihydrofluorescein in isolated liver and heart mitochondria was observed in the absence of ATP or K+ and therefore independent of K(ATP) channels. The response was blocked by stigmatellin, implying a role for the cytochrome bc1 complex (complex III). Diazoxide, though, did not increase hydrogen peroxide (H2O2) production (quantitatively measured with Amplex Red) in intact mitochondria, submitochondrial particles, or purified cytochrome bc1 complex. We confirmed that diazoxide inhibited succinate oxidation, but it also weakly stimulated state 4 respiration even in K+-free buffer, excluding a role for K(ATP) channels. Furthermore, we have shown previously that 5-hydroxydecanoate is partially metabolized, and we hypothesized that fatty acid metabolism may explain the ability of this putative mitochondrial K(ATP) channel blocker to inhibit diazoxide-induced flavoprotein fluorescence, commonly used as an assay of K(ATP) channel activity. Indeed, consistent with our hypothesis, we found that decanoate inhibited diazoxide-induced flavoprotein oxidation. Taken together, our data question the "mitochondrial K(ATP) channel" hypothesis of preconditioning. Diazoxide did not evoke superoxide (which dismutates to H2O2) from the respiratory chain by a direct mechanism, and the stimulatory effects of this compound on mitochondrial respiration and 2',7'-dichlorodihydrofluorescein oxidation were not due to the opening of K(ATP) channels. PMID- 16709572 TI - Ca2+ influx induced by protease-activated receptor-1 activates a feed-forward mechanism of TRPC1 expression via nuclear factor-kappaB activation in endothelial cells. AB - Thrombin activation of protease-activated receptor-1 induces Ca(2+) influx through store-operated cation channel TRPC1 in endothelial cells. We examined the role of Ca(2+) influx induced by the depletion of Ca(2+) stores in signaling TRPC1 expression in endothelial cells. Both thrombin and a protease-activated receptor-1-specific agonist peptide induced TRPC1 expression in human umbilical vein endothelial cells, which was coupled to an augmented store-operated Ca(2+) influx and increase in endothelial permeability. To delineate the mechanisms of thrombin-induced TRPC1 expression, we transfected in endothelial cells TRPC1 promoter-luciferase (TRPC1-Pro-Luc) construct containing multiple nuclear factor kappaB (NF-kappaB) binding sites. Co-expression of dominant negative IkappaBalpha mutant prevented the thrombin-induced increase in TRPC1 expression, indicating the key role of NF-kappaB activation in mediating the response. Using TRPC1 promoter-deletion mutant constructs, we showed that NF-kappaB binding sites located between -1623 and -871 in the TRPC1 5'-regulatory region were required for thrombin-induced TRPC1 expression. Electrophoretic mobility shift assay utilizing TRPC1 promoter-specific oligonucleotides identified that the DNA binding activities of NF-kappaB to NF-kappaB consensus sites were located in this domain. Supershift assays using NF-kappaB protein-specific antibodies demonstrated the binding of p65 homodimer to the TRPC1 promoter. Inhibition of store Ca(2+) depletion, buffering of intracellular Ca(2+), or down-regulation of protein kinase Calpha downstream of Ca(2+) influx all blocked thrombin-induced NF kappaB activation and the resultant TRPC1 expression in endothelial cells. Thus, Ca(2+) influx via TRPC1 is a critical feed-forward pathway responsible for TRPC1 expression. The NF-kappaB-regulated TRPC1 expression may be an essential mechanism of vascular inflammation and, hence, a novel therapeutic target. PMID- 16709573 TI - Oligomerization of the yeast alpha-factor receptor: implications for dominant negative effects of mutant receptors. AB - Oligomerization of G protein-coupled receptors is commonly observed, but the functional significance of oligomerization for this diverse family of receptors remains poorly understood. We used bioluminescence resonance energy transfer (BRET) to examine oligomerization of Ste2p, a G protein-coupled receptor that serves as the receptor for the alpha-mating pheromone in the yeast Saccharomyces cerevisiae, under conditions where the functional effects of oligomerization could be examined. Consistent with previous results from fluorescence resonance energy transfer (Overton, M. C., and Blumer, K. J. (2000) Curr. Biol. 10, 341 344), we detected efficient energy transfer between Renilla luciferase and a modified green fluorescent protein individually fused to truncated alpha-factor receptors lacking the cytoplasmic C-terminal tail. In addition, the low background of the BRET system allowed detection of significant, but less efficient, energy transfer between full-length receptors. The reduced efficiency of energy transfer between full-length receptors does not appear to result from different levels of receptor expression. Instead, attachment of fluorescent reporter proteins to the full-length receptors appears to significantly increase the distance between reporters. Mutations that were previously reported to block dimerization of truncated alpha-factor receptors reduce but do not completely eliminate BRET transfer between receptors. Dominant negative effects of mutant alleles of alpha-factor receptors appear to be mediated by receptor oligomerization since these effects are abrogated by introduction of additional mutations that reduce oligomerization. We find that heterodimers of normal and dominant negative receptors are defective in their ability to signal. Thus, signal transduction by oligomeric receptors appears to be a cooperative process requiring an interaction between functional monomers. PMID- 16709574 TI - JNK- and p38 kinase-mediated phosphorylation of Bax leads to its activation and mitochondrial translocation and to apoptosis of human hepatoma HepG2 cells. AB - Mitochondrial translocation of pro-apoptotic Bax prior to apoptosis is well established after treatment with many cell death stimulants or under apoptosis inducing conditions. The mechanism of mitochondrial translocation of Bax is, however, still unknown. The aim of this work was to investigate the mechanism of Bax activation and mitochondrial translocation to initiate apoptosis of human hepatoma HepG2 and porcine kidney LLC-PK1 cells exposed to various cell death agonists. Phosphorylation of Bax by JNK and p38 kinase activated after treatment with staurosporine, H(2)O(2), etoposide, and UV light was demonstrated by the shift in the pI value of Bax on two-dimensional gels and confirmed by metabolic labeling with inorganic [(32)P]phosphate in HepG2 cells. Specific inhibitors of JNK and p38 kinase significantly inhibited Bax phosphorylation and mitochondrial translocation and apoptosis of HepG2 cells. A specific small interfering RNA to MAPKK4 (the upstream protein kinase of JNK and p38 kinase) markedly decreased the levels of MAPKK4 and MAPKK3/6, blocked the activation of JNK or p38 kinase, and inhibited Bax phosphorylation. However, the negative control small interfering RNA did not cause these changes. Confocal microscopy of various Bax mutants showed differential rates of mitochondrial translocation of Bax before and after staurosporine treatment. Among the Bax mutants, T167D did not translocate to mitochondria after staurosporine exposure, suggesting that Thr(167) is a potential phosphorylation site. In conclusion, our results demonstrate, for the first time, that Bax is phosphorylated by stress-activated JNK and/or p38 kinase and that phosphorylation of Bax leads to mitochondrial translocation prior to apoptosis. PMID- 16709575 TI - Dealing with innovation and costs in orthopedics: a conversation with Dane Miller. Interview by Lawton R Burns. AB - Rob Burns talks with Dane Miller, former CEO of Biomet, about challenges posed by new technology in the orthopedic devices area. One key challenge is the rising cost and use of orthopedic devices at a time when providers are facing decreased profitability and reimbursement for orthopedic services. Another challenge is the long-term time horizon needed to gauge product success that contrasts with payers' and providers' short-term horizon. A third challenge is heightened governmental scrutiny of device makers' relationships with orthopedic surgeons. This interview was conducted before Miller left Biomet in March 2006. PMID- 16709576 TI - Shoot development and extension of Quercus serrata saplings in response to insect damage and nutrient conditions. AB - BACKGROUND AND AIMS: Plants have the ability to compensate for damage caused by herbivores. This is important to plant growth, because a plant cannot always avoid damage, even if it has developed defence mechanisms against herbivores. In previous work, we elucidated the herbivory-induced compensatory response of Quercus (at both the individual shoot and whole sapling levels) in both low- and high-nutrient conditions throughout one growing season. In this study, we determine how the compensatory growth of Quercus serrata saplings is achieved at different nutrient levels. METHODS: Quercus serrata saplings were grown under controlled conditions. Length, number of leaves and percentage of leaf area lost on all extension units (EUs) were measured. KEY RESULTS: Both the probability of flushing and the length of subsequent EUs significantly increased with an increase in the length of the parent EU. The probability of flushing increased with an increase in leaf damage of the parent EU, but the length of subsequent EUs decreased. This indicates that EU growth is fundamentally regulated at the individual EU level. The probabilities of a second and third flush were significantly higher in plants in high-nutrient soil than those in low-nutrient soil. The subsequent EUs of damaged saplings were also significantly longer at high-nutrient conditions. CONCLUSIONS: An increase in the probability of flushes in response to herbivore damage is important for damaged saplings to produce new EUs; further, shortening the length of EUs helps to effectively reproduce foliage lost by herbivory. The probability of flushing also varied according to soil nutrient levels, suggesting that the compensatory growth of individual EUs in response to local damage levels is affected by the nutrients available to the whole sapling. PMID- 16709577 TI - Cancer prevention: strategy for the future. PMID- 16709578 TI - Dietary effects of soy isoflavones daidzein and genistein on 7,12 dimethylbenz[a]anthracene-induced mammary mutagenesis and carcinogenesis in ovariectomized Big Blue transgenic rats. AB - The major constituents of isoflavones daidzein (DZ) and genistein (GE) interact with the and estrogen receptors in several tissues including mammary tissues. In this study, we used ovariectomy (OVX) to model menopause and determined the effects of DZ, GE or 17beta-estradiol (E(2)) exposures on chemically induced mutagenesis and carcinogenesis in the mammary glands of female Big Blue transgenic rats. The rats were fed control diet containing the isoflavones and E(2) and treated with a single oral dose of 7,12-dimethylbenz[a]anthracene (DMBA) at PND50. Animals were euthanized at 16 or 20 weeks post-carcinogen treatment to assess mutant frequencies (MFs) and histopathological parameters, respectively. The isoflavones or E(2) supplementation alone resulted in the lac I MFs that were not significantly different from the MFs measured in rats fed the control diet alone. DMBA exposure, however, induced significant increases in the lac I MFs in the mammary tissues of both OVX and INT rats and Hprt MFs in spleen lymphocytes (P < 0.01). In general, feeding the isoflavones or E(2) did not cause any significant changes in DMBA-induced mutagenicity in the mammary tissues. However, feeding the isoflavone mixture (daidzein + genistein; DZG) resulted in a significant reduction in the DMBA-induced lac I MFs (P < 0.05). Cell proliferation as measured by PCNA immunohistochemistry was increased in both OVX and INT rats exposed to DMBA as compared with rats fed control diet (P < 0.05). Mammary histology indicated that hyperplasia was induced in most of the treatment groups including control. Although DMBA did not induce mammary tumors in the OVX rats, adenoma and adenocarcinoma were detected in the mammary glands of INT rats. PMID- 16709579 TI - Subarachnoid haemorrhage in patients > or = 75 years: clinical course, treatment and outcome. PMID- 16709580 TI - Schooling mediates brain reserve in Alzheimer's disease: findings of fluoro-deoxy glucose-positron emission tomography. AB - BACKGROUND: Functional imaging studies report that higher education is associated with more severe pathology in patients with Alzheimer's disease, controlling for disease severity. Therefore, schooling seems to provide brain reserve against neurodegeneration. OBJECTIVE: To provide further evidence for brain reserve in a large sample, using a sensitive technique for the indirect assessment of brain abnormality (18F-fluoro-deoxy-glucose-positron emission tomography (FDG-PET)), a comprehensive measure of global cognitive impairment to control for disease severity (total score of the Consortium to Establish a Registry for Alzheimer's Disease Neuropsychological Battery) and an approach unbiased by predefined regions of interest for the statistical analysis (statistical parametric mapping (SPM)). METHODS: 93 patients with mild Alzheimer's disease and 16 healthy controls underwent 18F-FDG-PET imaging of the brain. A linear regression analysis with education as independent and glucose utilisation as dependent variables, adjusted for global cognitive status and demographic variables, was conducted in SPM2. RESULTS: The regression analysis showed a marked inverse association between years of schooling and glucose metabolism in the posterior temporo occipital association cortex and the precuneus in the left hemisphere. CONCLUSIONS: In line with previous reports, the findings suggest that education is associated with brain reserve and that people with higher education can cope with brain damage for a longer time. PMID- 16709581 TI - Polyneuropathy dysfunction scores. PMID- 16709582 TI - Neuromyelitis optica and anti-aquaporin-4 antibodies: widening the clinical phenotype. PMID- 16709584 TI - First Latin American position paper on the pharmacological treatment of rheumatoid arthritis. AB - BACKGROUND: Rheumatoid arthritis (RA) is a chronic, systemic inflammatory disease that involves synovial joints, resulting in severe dysfunction or burden for individual patients, families and society. Latin American Rheumatology Associations have acknowledged its relevance and recognized multiple limitations for its diagnosis and treatment in Latin America and the Caribbean. This document underscores issues regarding the impact and relevance of this disease in these countries. OBJECTIVES: To develop a consensus document that may unify and guide the pharmacological management of RA in Latin America and the Caribbean. METHODS: An Executive Committee appointed by the Epidemiology, Rheumatoid Arthritis and Radiology Committees of Pan-American League of Association for Rheumatology (PANLAR), held a meeting at Lisbon in May 2003. The goal was to establish a task force for the development of a Latin American consensus on the management of RA. Efforts focused on the problems encountered in the region regarding the availability of appropriate treatment for RA and the development of treatment guidelines for clinical practice. A secondary objective was the diffusion of the consensus conclusions and recommendations in participating countries. RESULTS: Six major issues were identified for discussion by six working groups. All Latin American Rheumatology Associations registered in PANLAR were invited to participate in the consensus. PANLAR members were well-represented in each group. Coordinators identified essential literature to be reviewed, analysed, and electronically discussed before the consensus meeting. CONCLUSIONS: The consensus' results and recommendations of this effort to delineate RA management in Latin America are contained in this article, which has been reviewed by participant societies and authors during 2004/2005 and endorsed by PANLAR. PMID- 16709585 TI - Comparative gene marker selection suite. AB - MOTIVATION: An important step in analyzing expression profiles from microarray data is to identify genes that can discriminate between distinct classes of samples. Many statistical approaches for assigning significance values to genes have been developed. The Comparative Marker Selection suite consists of three modules that allow users to apply and compare different methods of computing significance for each marker gene, a viewer to assess the results, and a tool to create derivative datasets and marker lists based on user-defined significance criteria. AVAILABILITY: The Comparative Marker Selection application suite is freely available as a GenePattern module. The GenePattern analysis environment is freely available at http://www.broad.mit.edu/genepattern. PMID- 16709586 TI - A model diagram layout extension for SBML. AB - MOTIVATION: Since the knowledge about processes in living cells is increasing, modelling and simulation techniques are used to get new insights into these complex processes. During the last few years, the SBML file format has gained in popularity and support as a means of exchanging model data between the different modelling and simulation tools. In addition to specifying the model as a set of equations, many modern modelling tools allow the user to create and to interact with the model in the form of a reaction graph. Unfortunately, the SBML file format does not provide for the storage of this graph data along with the mathematical description of the model. RESULTS: Therefore, we developed an extension to the SBML file format that makes it possible to store such layout information which describes position and size of objects in the graphical representation. AVAILABILITY: The complete specification can be found on (http://projects.villa-bosch.de/bcb/sbml/ (SBML Layout Extension documentation, 2005). Additionally, a complete implementation exists as part of libSBML (2006, http://www.sbml.org/software/libsbml/). PMID- 16709587 TI - INFO-RNA--a fast approach to inverse RNA folding. AB - MOTIVATION: The structure of RNA molecules is often crucial for their function. Therefore, secondary structure prediction has gained much interest. Here, we consider the inverse RNA folding problem, which means designing RNA sequences that fold into a given structure. RESULTS: We introduce a new algorithm for the inverse folding problem (INFO-RNA) that consists of two parts; a dynamic programming method for good initial sequences and a following improved stochastic local search that uses an effective neighbor selection method. During the initialization, we design a sequence that among all sequences adopts the given structure with the lowest possible energy. For the selection of neighbors during the search, we use a kind of look-ahead of one selection step applying an additional energy-based criterion. Afterwards, the pre-ordered neighbors are tested using the actual optimization criterion of minimizing the structure distance between the target structure and the mfe structure of the considered neighbor. We compared our algorithm to RNAinverse and RNA-SSD for artificial and biological test sets. Using INFO-RNA, we performed better than RNAinverse and in most cases, we gained better results than RNA-SSD, the probably best inverse RNA folding tool on the market. AVAILABILITY: www.bioinf.uni freiburg.de?Subpages/software.html. PMID- 16709588 TI - Combo: a whole genome comparative browser. AB - SUMMARY: Combo is a comparative genome browser that provides a dynamic view of whole genome alignments along with their associated annotations. Combo provides two different visualization perspectives. The perpendicular (dot plot) view provides a dot plot of genome alignments synchronized with a display of genome annotations along each axis. The parallel view displays two genome annotations horizontally, synchronized through a panel displaying local alignments as trapezoids. Users can zoom to any resolution, from whole chromosomes to individual bases. They can select, highlight and view detailed information from specific alignments and annotations. Combo is an organism agnostic and can import data from a variety of file formats. AVAILABILITY: Combo is integrated as part of the Argo Genome Browser which also provides single-genome browsing and editing capabilities. Argo is written in Java, runs on multiple platforms and is freely available for download at http://www.broad.mit.edu/annotation/argo/. PMID- 16709589 TI - Independent component analysis-based penalized discriminant method for tumor classification using gene expression data. AB - MOTIVATION: Microarrays are capable of determining the expression levels of thousands of genes simultaneously. One important application of gene expression data is classification of samples into categories. In combination with classification methods, this technology can be useful to support clinical management decisions for individual patients, e.g. in oncology. Standard statistic methodologies in classification or prediction do not work well when the number of variables p (genes) far too exceeds the number of samples n. So, modification of existing statistical methodologies or development of new methodologies is needed for the analysis of microarray data. RESULTS: This paper proposes a new method for tumor classification using gene expression data. In this method, we first employ independent component analysis to model the gene expression data, then apply optimal scoring algorithm to classify them. Further speaking, this approach can first make full use of the high-order statistical information contained in the gene expression data. Second, this approach also employs regularized regression models to handle the situation of large numbers of correlated predictor variables. Finally, the predictive models are developed for classifying tumors based on the entire gene expression profile. To show the validity of the proposed method, we apply it to classify four DNA microarray datasets involving various human normal and tumor tissue samples. The experimental results show that the method is efficient and feasible. AVAILABILITY: Matlab scripts are available on request. PMID- 16709590 TI - TRFMA: a web-based tool for terminal restriction fragment length polymorphism analysis based on molecular weight. AB - TRFMA provides a Web environment for analyzing T-RFLP results based on molecular weights of the fragments, rather than the numbers of nucleotides, to increase accuracy. The 16S rRNA data are saved as an XML file containing around 650 sequences (light version) and a MySQL database containing around 50 000 sequences (full version), which are connected to Web server via PHP5 and manipulated on an Internet browser. AVAILABILITY: TRFMA is freely available at http://myamagu.dent.kyushu-u.ac.jp/bioinformatics/trfma/index.html and can be downloaded from the same site. PMID- 16709592 TI - Toxicity, stability and pharmacokinetics of amphotericin B in immunomodulator tuftsin-bearing liposomes in a murine model. AB - OBJECTIVES: In the present study we evaluated the pharmacokinetics and toxicity of amphotericin B in immunomodulator tuftsin-loaded liposomes in a murine model. METHODS: Stability of amphotericin B liposomes was tested by incubating one volume of liposomal formulations of amphotericin B with nine volumes of serum. The pharmacokinetics of amphotericin B in Candida albicans-infected mice treated with conventional and tuftsin-loaded amphotericin B liposomes was evaluated over a period of 24 h. In vitro toxicity of amphotericin B deoxycholate, as well as amphotericin B liposomes, was tested by incubation with human erythrocytes for 1 h at 37 degrees C. To assess amphotericin B-induced in vivo toxicity, BALB/c mice were injected with three doses of amphotericin B deoxycholate, as well as amphotericin B liposomal formulations on days 1, 2 and 3 post C. albicans infection. Blood from treated mice was taken by retro-orbital puncture to test renal function parameters such as serum creatinine and urea. RESULTS: In vitro stability studies revealed that tuftsin-bearing amphotericin B liposomes released only 11% of the total liposomal amphotericin B in the serum, while it was found to be 19% from identical tuftsin-free amphotericin B liposomes. Both tuftsin loaded as well as tuftsin-free liposomal formulations of amphotericin B induced approximately 20% haemolysis of erythrocytes at a dose of 40 mg/L, while the same amount of drug in amphotericin B deoxycholate caused 100% lysis of the erythrocytes. Pharmacokinetic studies revealed that subsequent to administration of various formulations of amphotericin B, there was 32 mg/L amphotericin B in the systemic circulation of mice treated with tuftsin-bearing amphotericin B liposomes, while it was 25 mg/L for amphotericin B liposomes, 4 h post drug administration. In vivo toxicity studies demonstrated that the amphotericin B deoxycholate formulation induced elevations in serum creatinine (approximately 300% of control) and blood urea (approximately 380% of control) values, while these values were substantially less (blood urea approximately 150% of control and serum creatinine approximately 210% of control) in the animals treated with the tuftsin-loaded amphotericin B liposomal formulation. Further, the administration of amphotericin B deoxycholate (1 mg/kg) in BALB/c mice at a dose of 1 mg/kg body weight led to the accumulation of 18.6 +/- 5.25 g/kg (of amphotericin B) in kidneys. On the other hand, administration of liposomal amphotericin B and tuftsin-bearing liposomal amphotericin B at a dose of 5 mg/kg body weight resulted in accumulation of 8.8 +/- 2.0 and 4.0 +/- 1.6 g/kg of amphotericin B, respectively, in the kidneys of treated animals. CONCLUSIONS: Co administration of immunomodulator tuftsin along with liposomal formulations of amphotericin B successfully minimizes toxicity, as well as other side effects of the drug. Interestingly, tuftsin also increased the stability of liposomal amphotericin B. Superior efficacy, reliable safety and favourable pharmacodynamics of tuftsin-loaded amphotericin B liposomes suggest their potential therapeutic value in the management of fungal infections. PMID- 16709591 TI - Context-specific infinite mixtures for clustering gene expression profiles across diverse microarray dataset. AB - MOTIVATION: Identifying groups of co-regulated genes by monitoring their expression over various experimental conditions is complicated by the fact that such co-regulation is condition-specific. Ignoring the context-specific nature of co-regulation significantly reduces the ability of clustering procedures to detect co-expressed genes due to additional 'noise' introduced by non-informative measurements. RESULTS: We have developed a novel Bayesian hierarchical model and corresponding computational algorithms for clustering gene expression profiles across diverse experimental conditions and studies that accounts for context specificity of gene expression patterns. The model is based on the Bayesian infinite mixtures framework and does not require a priori specification of the number of clusters. We demonstrate that explicit modeling of context-specificity results in increased accuracy of the cluster analysis by examining the specificity and sensitivity of clusters in microarray data. We also demonstrate that probabilities of co-expression derived from the posterior distribution of clusterings are valid estimates of statistical significance of created clusters. AVAILABILITY: The open-source package gimm is available at http://eh3.uc.edu/gimm. PMID- 16709593 TI - Antifungal and antimycobacterial activity of new imidazole and triazole derivatives. A combined experimental and computational approach. AB - OBJECTIVES: To synthesize new antimycobacterial and antifungal drugs that act by binding to sterol 14alpha-demethylase (14DM) and to characterize the drug-target protein interactions using computer-based molecular simulations. METHODS: Different series of imidazole and triazole derivatives having an azomethine linkage to pyridine 2-carboxamidrazone were designed and synthesized. Molecular dynamic simulations of the sterol 14DM (a mixed-function oxidase involved in sterol synthesis in eukaryotic and prokaryotic organisms) complexed with new azole derivatives have been performed to both qualify and quantify the protein ligand interactions. MICs of the compounds were evaluated by reference assay and by the recently developed Microdilution Resazurin Assay (MRA). RESULTS: Halogenated derivatives showed good activity, with an MIC90 of 1 mg/L against 33 Candida spp. clinical strains; most compounds also had inhibitory activity against Mycobacterium tuberculosis reference and clinical strains, with MICs in the range 4-64 mg/L. Molecular modelling investigations showed that the active new compounds may interact at the active site of both the fungal and the mycobacterial cytochrome P450-dependent sterol-14alpha-demethylase and that the calculated binding free energy values are in agreement with the corresponding MIC values. CONCLUSIONS: The combined experimental and computational approach can be helpful in targeted drug design, thus yielding valuable information for the synthesis and prediction of activity of a second generation of inhibitors. PMID- 16709594 TI - Correlation between apramycin and gentamicin use in pigs and an increasing reservoir of gentamicin-resistant Escherichia coli. AB - OBJECTIVES: Resistance towards the veterinary drug apramycin can be caused by the aac(3)-IV gene, which also confers resistance towards the important human antibiotic gentamicin. The objectives of this study were to investigate the temporal occurrence and the genetic background of apramycin and gentamicin resistance in Escherichia coli strains from pork, healthy pigs and diagnostic submissions from pigs and to investigate potential relationships to the use of apramycin and gentamicin at farm and national levels. METHODS: Data on Danish E. coli isolates from healthy pigs (indicator bacteria), diagnostic submissions from pigs (clinical isolates) and pork were obtained from the national surveillance of antimicrobial resistance and from routine diagnostic laboratories. Antimicrobial consumption data were obtained from the Danish Medicines Agency (1997-2000) and from the VetStat database (2001-2004). The genetic background for gentamicin resistance was investigated by PCR. Relationships between antimicrobial usage and resistance were analysed by chi2 test and logistic regression. RESULTS: At the farm level, the occurrence of apramycin/gentamicin cross-resistance was correlated to the use of apramycin (P < 0.001). At the national level, occurrence of apramycin/gentamicin cross-resistance in clinical E. coli O149 isolates was significantly correlated with the amounts and duration of apramycin use. The aac(3)-IV gene was detected in all tested cross-resistant isolates. CONCLUSIONS: Apramycin consumption at farm level is most probably driving the increasing occurrence of apramycin/gentamicin cross-resistant [aac(3)-IV positive] E. coli in diseased pigs and healthy finishers at slaughter. The duration of use and amounts used both had a significant effect on the prevalence of apramycin/gentamicin cross-resistance in diseased weaning pigs at the national level. PMID- 16709596 TI - Gene regulation of neurokinin B and its receptor NK3 in late pregnancy and pre eclampsia. AB - Elevated circulating levels of the tachykinin, neurokinin B (NKB), have been observed in women with pre-eclampsia during the third trimester of pregnancy. Currently, the molecular mechanisms responsible for these increased levels remain unknown. To understand the molecular regulation, we have compared the differences in gene expression of the tachykinins and their receptors in control and pre eclamptic placentae and the responses of the TAC3 gene encoding NKB to proposed physiological triggers of pre-eclampsia including hypoxia and oxidative stress using real-time quantitative PCR. We have determined the placenta to be the main site of TAC3 expression with levels 2.6-fold higher than the brain. TAC3 expression was found to be significantly higher in pre-eclamptic placenta (1.7 fold, P < 0.05) than in normal controls. No evidence was found that hypoxia and oxidative stress were responsible for increases in TAC3 expression. In rat placenta, a longitudinal study in normal late pregnancy was associated with a significant down-regulation of the NKB/NK3 ligand-receptor pair (P < 0.05). The present data suggest that the increased placental expression of TAC3 is part of the mechanism leading to the increased circulating levels of NKB in pre eclampsia. PMID- 16709595 TI - Heart failure, chronic diuretic use, and increase in mortality and hospitalization: an observational study using propensity score methods. AB - AIMS: Non-potassium-sparing diuretics are commonly used in heart failure (HF). They activate the neurohormonal system, and are potentially harmful. Yet, the long-term effects of chronic diuretic use in HF are largely unknown. We retrospectively analysed the Digitalis Investigation Group (DIG) data to determine the effects of diuretics on HF outcomes. METHODS AND RESULTS: Propensity scores for diuretic use were calculated for each of the 7788 DIG participants using a non-parsimonious multivariable logistic regression model, and were used to match 1391 (81%) no-diuretic patients with 1391 diuretic patients. Effects of diuretics on mortality and hospitalization at 40 months of median follow-up were assessed using matched Cox regression models. All-cause mortality was 21% for no-diuretic patients and 29% for diuretic patients [hazard ratio (HR) 1.31; 95% confidence interval (CI) 1.11-1.55; P = 0.002]. HF hospitalizations occurred in 18% of no-diuretic patients and 23% of diuretic patients (HR 1.37; 95% CI 1.13-1.65; P = 0.001). CONCLUSION: Chronic diuretic use was associated with increased long-term mortality and hospitalizations in a wide spectrum of ambulatory chronic systolic and diastolic HF patients. The findings of the current study challenge the wisdom of routine chronic use of diuretics in HF patients who are asymptomatic or minimally symptomatic without fluid retention, and are on complete neurohormonal blockade. These findings, based on a non-randomized design, need to be further studied in randomized trials. PMID- 16709597 TI - Signal transducer and activator of transcription (stat) binding sites but not stat3 are required for fasting-induced transcription of agouti-related protein messenger ribonucleic acid. AB - Energy homeostasis depends on the regulation of hypothalamic neurons by leptin, an adipocyte hormone whose circulating levels communicate body energy stores. Leptin activates the transcription factor signal transducer and activator of transcription 3 (Stat3) in hypothalamic neurons, including neuronal subtypes producing Agouti-related protein (Agrp), a neuropeptide that stimulates feeding. Previous studies have suggested a model in which high levels of Agrp transcription during fasting represent a default state that is actively repressed by phospho-Stat3 induced by leptin signaling in the fed state. We identify putative Stat3 binding elements in the Agrp promoter that have been highly conserved during vertebrate evolution. Using a reporter assay in transgenic mice that faithfully recapitulates normal regulation of Agrp, we show that these sites are required, but in a way opposite to that predicted by the existing model: mutation of the sites leads to a default state characterized by a low level of Agrp transcription and insensitivity to fasting. We also find that removing activatable Stat3 from Agrp neurons has no detectable effect on steady-state levels of Agrp mRNA in the fed or fasted state. These results suggest a new model for transcriptional regulation of orexigenic neuropeptides in which the default level of expression is low in the fed state, and transcriptional activation in response to fasting is mediated by factors other than Stat3. PMID- 16709598 TI - Role and regulation of nodal/activin receptor-like kinase 7 signaling pathway in the control of ovarian follicular atresia. AB - Although the role of the TGF beta superfamily members in the regulation of ovarian folliculogenesis has been extensively studied, their involvement in follicular atresia is not well understood. In the present study, we have demonstrated for the first time that Nodal, a member of the TGF beta superfamily, is involved in promoting follicular atresia as evidenced by the following: 1) colocalization of Nodal and its type I receptor Activin receptor-like kinase 7 (ALK7) proteins in the granulosa cells was only observed in atretic antral follicles, whereas they were present in theca cells and granulosa cells of healthy follicles, respectively; 2) addition of recombinant Nodal or overexpression of Nodal by adenoviral infection induced apoptosis of otherwise healthy granulosa cells; 3) constitutively active ALK7 (ALK7-ca) overexpression mimicked the function of Nodal in the induction of granulosa cell apoptosis. Furthermore, overexpression of Nodal or ALK7-ca increased phosphorylation and nuclear translocation of Smad2, decreased X-linked inhibitor of apoptotic proteins (Xiap) expression at both mRNA and protein level and phospho-Akt content, as well as triggered mitochondrial release of death proteins Smac/DIABLO, Omi/HtrA2, and cytochrome c in the granulosa cells. Dominant negative Smad2 significantly attenuated ALK7-ca-induced down-regulation of Xiap and thus rescued granulosa cells from undergoing apoptosis. In addition, whereas up-regulation of Xiap significantly attenuated ALK7-ca-induced apoptosis, down regulation of Xiap sensitized granulosa cells to ALK7-ca-induced apoptosis. Furthermore, ALK7-ca-induced apoptosis was significantly attenuated by forced expression of activated Akt, and Akt rescued granulosa cells from undergoing apoptosis via proteasome-mediated ALK7 degradation. Taken together, Nodal plays an atretogenic role in the ovary where it induces granulosa cell apoptosis through activation of Smad2, down-regulation of the key survival molecules Xiap and phospho-Akt, as well as the activation of mitochondrial death pathway. PMID- 16709599 TI - Dosage-sensitive sex reversal adrenal hypoplasia congenita critical region on the X chromosome, gene 1 (DAX1) (NR0B1) and small heterodimer partner (SHP) (NR0B2) form homodimers individually, as well as DAX1-SHP heterodimers. AB - Dosage-sensitive sex reversal adrenal hypoplasia congenita critical region on the X chromosome, gene 1 (DAX1) (NR0B1), and small heterodimer partner (SHP) (NR0B2) are atypical nuclear receptor superfamily members that function primarily as corepressors through heterodimeric interactions with other nuclear receptors. Mutations in DAX1 cause adrenal hypoplasia congenita, and mutations in SHP lead to mild obesity and insulin resistance, but the mechanisms are unclear. We investigated the existence and subcellular localization of DAX1 and SHP homodimers and the dynamics of homodimerization. We demonstrated DAX1 homodimerization in the nucleus and cytoplasm, and dissociation of DAX1 homodimers upon heterodimerization with steroidogenic factor 1 (SF1) or ligand activated estrogen receptor-alpha (ERalpha). DAX1 homodimerization involved an interaction between its amino and carboxy termini involving its LXXLL motifs and activation function (AF)-2 domain. We observed SHP homodimerization in the nucleus of mammalian cells and showed dissociation of SHP homodimers upon heterodimerization with ligand-activated ERalpha. We observed DAX1-SHP heterodimerization in the nucleus of mammalian cells and demonstrated the involvement of the LXXLL motifs and AF-2 domain of DAX1 in this interaction. We further demonstrate heterodimerization of DAX1 with its alternatively spliced isoform, DAX1A. This is the first evidence of homodimerization of individual members of the unusual NR0B nuclear receptor family and heterodimerization between its members. Our results suggest that DAX1 forms antiparallel homodimers through the LXXLL motifs and AF-2 domain. These homodimers may function as holding reservoirs in the absence of heterodimeric partners. The formation of DAX1 and SHP homodimers and DAX1-SHP and DAX1-DAX1A heterodimers suggests the possibility of novel functions independent of their coregulator roles, suggesting additional complexity in the molecular mechanisms of DAX1 and SHP action. PMID- 16709600 TI - Differential expression of FOXO1 and FOXO3a confers resistance to oxidative cell death upon endometrial decidualization. AB - The integrity of the feto-maternal interface is critical for survival of the conceptus. This interface, consisting of the maternal decidua and the invading placental trophoblast, is exposed to profound changes in oxygen tension during pregnancy. We demonstrate that human endometrial stromal cells become extraordinarily resistant to oxidative stress-induced apoptosis upon decidualization in response to cAMP and progesterone signaling. This differentiation process is associated with the induction of the forkhead transcription factor FOXO1, which in turn increases the expression of the mitochondrial antioxidant manganese superoxide dismutase. However, silencing of FOXO1 did not increase the susceptibility of decidualized cells to oxidative cell death. Comparative analysis demonstrated that hydrogen peroxide, a source of free radicals, strongly induces FOXO3a mRNA and protein expression in undifferentiated human endometrial stromal cells but not in decidualized cells. Expression of a constitutively active FOXO3a mutant elicited apoptosis in decidualized cells. Furthermore, silencing of endogenous FOXO3a in undifferentiated cells abrogated apoptosis induced by hydrogen peroxide. These results suggest that the induction of FOXO1 may enhance the ability of decidualized cells to prevent oxidative damage while the simultaneous repression of FOXO3a expression disables the signaling pathway responsible for oxidative cell death. The differential regulation of FOXO expression provides the decidua with a robust system capable of coping with prolonged episodes of oxidative stress during pregnancy. PMID- 16709601 TI - Biological effect of a novel mutation in the third leucine-rich repeat of human luteinizing hormone receptor. AB - A novel heterozygous mutation A340T leading to the substitution of Phe for the conserved amino acid Ile114 was identified by nucleotide sequencing of the human LH/chorionic gonadotropin receptor (hLHR) of a patient with Leydig cell hypoplasia. This mutation is located in the third leucine-rich repeat in the ectodomain of the hLHR. In vitro expression studies demonstrated that this mutation results in reduced ligand binding and signal transduction of the receptor. Studies of hLHR constructs in which various amino acids were substituted for the conserved Ile114 showed that receptor activity is sensitive to changes in size, shape, and charge of the side chain. A homology model of the wild-type hLHR ectodomain was made, illustrating the packing of conserved hydrophobic side chains in the protein core. Substitution of Ile114 by Phe might disrupt intermolecular contacts between hormone and receptor. This mutation might also affect an LHR-dimer interaction. Thus, the I114F mutation reduces ligand binding and signal transduction by the hLHR, and it is partially responsible for Leydig cell hypoplasia in the patient. PMID- 16709602 TI - A novel follicle-stimulating hormone-induced G alpha h/phospholipase C-delta1 signaling pathway mediating rat sertoli cell Ca2+-influx. AB - FSH is known to activate Gs/cAMP signaling pathway in Sertoli cells (SCs) to support spermatogenesis. However, the molecular mechanism of FSH-induced Gs/cAMP independent Ca2+-influx in SCs is not clear. In this study, FSH indeed induced an immediate and dose-dependent intracellular Ca2+-elevation in rat SCs. In the presence of EDTA (2.5 mm) or in the absence of extracellular Ca2+, the FSH induced intracellular Ca2+-elevation was abolished. The confocal microscopic observation of Ca2+ image revealed that the SC cellular Ca2+ level was gradually increased after 50 sec of FSH treatment. Dantrolene, a blocker of intracellular Ca2+ release, did not affect this FSH-induced intracellular Ca2+ elevation. The pretreatment of rat SCs with phosphatidylinositol-phospholipase C (PLC)-specific inhibitor, U73122 (3 and 10 microm), inhibited the FSH-induced Ca2+-influx in a dose-dependent manner, but treatment with Gs-specific inhibitor, NF449 (0.1 and 0.3 microm), did not. On the other hand, the activation of G alpha h was immediately induced by FSH in the rat SCs within 5 sec of treatment. The translocation of PLC-delta1 from cytosol to cell membrane and the formation of G alpha h /PLC-delta1 complexes occurred within 5 and 10 sec, respectively, of FSH exposure. The intracellular inositol 1,4,5-triphosphate (IP3) production was also detected after 30 sec of FSH treatment. The synthetic peptide of PLC-delta1 (TIPWNSLKQGYRHVHLL), not Gs inhibitor, predominantly inhibited the FSH-induced PLC-delta1 translocation, formation of G alpha h /PLC-delta1 complex, intracellular IP3 production, and Ca2+ influx. In contrast, the peptide did not interfere with FSH-induced intracellular cAMP accumulation. In conclusion, the FSH-induced immediate Ca2+ influx is unambiguously mediated by an alternative G alpha h /PLC-delta1/IP3 pathway that is distinct from the Gs/cAMP pathway in rat SCs. PMID- 16709603 TI - The role of three-dimensional ultrasonography in polycystic ovary syndrome. AB - The 2003 Rotterdam diagnostic criteria for polycystic ovary syndrome (PCOS) support the objective role of ultrasound in defining the appearance of the PCO, but there are significant limitations of these new guidelines from an ultrasound perspective that must be considered. Three-dimensional (3D) ultrasound provides a new method for the objective quantitative assessment of follicle count, ovarian volume, stromal volume and blood flow within the ovary as a whole. Since the introduction of 3D ultrasonography, there have been increasing publications on PCOS, each addressing different areas and reporting different results. This review critically examines these studies in an attempt to clarify the evidence to date and thereby establish the current role of 3D ultrasonography in PCOS. PMID- 16709604 TI - Decrease of both stem cell factor and clusterin mRNA levels in testicular biopsies of azoospermic patients with constitutive or idiopathic but not acquired spermatogenic failure. AB - BACKGROUND: Sertoli cells nurse germ cells during spermatogenesis, and alterations of Sertoli cell functions have been suggested in cases of spermatogenic failures. METHODS: In this work, we measured stem cell factor (SCF) and clusterin mRNA levels, by quantitative RT-PCR, in RNA extracted from testicular biopsies of 49 azoospermic patients classified according to testicular histology as having normal spermatogenesis or spermatogenic failure. RESULTS: When related to the percentage of Sertoli cells counted on a histological section of a neighbouring tissue sample, SCF and clusterin mRNA levels were significantly lower in the 'spermatogenic failure' group compared with the control group (P = 0.0297 and P = 0.0043, respectively). These levels were also significantly lower in the cases of 'constitutive' (cryptorchidism and Yq microdeletion) and 'idiopathic' spermatogenic failures when compared with the control group; conversely, they were not significantly decreased in the group with 'acquired spermatogenic failure' (orchitis, testicular traumatism, chemoradiotherapy and varicocele). CONCLUSIONS: These data further demonstrate an alteration of Sertoli cell functions in some human spermatogenic failures and suggest that a lack of Sertoli cell maturation may be involved in cases of constitutive or idiopathic spermatogenic failures. PMID- 16709606 TI - Do carers' needs assessments make a difference? Results from the Forget me not study. PMID- 16709605 TI - Clinical relevance of low serum vitamin B12 concentrations in older people: the Banbury B12 study. AB - BACKGROUND: low vitamin B12 concentrations are common in older people, but the clinical relevance of biochemical evidence of vitamin B12 deficiency in the absence of anaemia is uncertain. OBJECTIVE: to examine associations of cognitive impairment, depression and neuropathy with blood measurements of vitamin B12 and folate status in older people. DESIGN: cross-sectional study in general practice in Banbury, England. PARTICIPANTS: a total of 1,000 individuals aged 75 years or older living in the community. RESULTS: low vitamin B12 concentrations were identified in 13% of older people and were associated with memory impairment and depression. After adjustment for age, sex and smoking, individuals with vitamin B12 or holotranscobalamin (holoTC) in the bottom compared with top quartiles had a 2-fold risk (OR = 2.17; 95% CI 1.11-4.27) and a 3-fold risk (OR = 3.02; 95% CI 1.31-6.98) of cognitive impairment, respectively. Low vitamin B12 status was also associated with missing ankle tendon jerks but not with depression. Treatment with vitamin B12 for 3 months corrected the biochemical abnormalities but had no effect on any of the clinical measurements. CONCLUSIONS: low vitamin B12 concentrations are associated with cognitive impairment and missing ankle tendon jerks in older people in the absence of anaemia. Large-scale trials of vitamin B12 supplementation are required to assess the clinical significance of these associations. PMID- 16709607 TI - Endogenous interleukin-10 protects against hepatic steatosis but does not improve insulin sensitivity during high-fat feeding in mice. AB - Several studies have demonstrated an association in humans between plasma levels or production capacity of the antiinflammatory cytokine IL-10 and insulin sensitivity. The aim of our study was to investigate the protective role of endogenous IL-10 availability in the development of diet-induced insulin resistance. We compared parameters of glucose and lipid metabolism between IL-10( /-) mice and wild-type (wt) mice fed a high-fat diet for 6 wk. This diet has previously been shown to induce steatosis and insulin resistance. After 6 wk on the high-fat diet, no differences in body weight, basal metabolism (measured by indirect calorimetry), or plasma levels of glucose, triglycerides, or cholesterol were observed between IL-10(-/-) and wt mice. Nonetheless, in IL-10(-/-) mice, plasma free fatty acid levels were 75% increased compared with wt mice after overnight fasting (P < 0.05). In addition, hepatic triglyceride content was 54% increased in IL-10(-/-) mice (P < 0.05). During a hyperinsulinemic euglycemic clamp, no differences were observed in whole-body or hepatic insulin sensitivity between both groups. We conclude that basal IL-10 production protects against hepatic steatosis but does not improve hepatic or whole-body insulin sensitivity, during high-fat feeding. PMID- 16709608 TI - Tissue-specific thyroid hormone deprivation and excess in monocarboxylate transporter (mct) 8-deficient mice. AB - Mutations of the X-linked thyroid hormone (TH) transporter (monocarboxylate transporter, MCT8) produce in humans unusual abnormalities of thyroid function characterized by high serum T3 and low T4 and rT3. The mechanism of these changes remains obscure and raises questions regarding the regulation of intracellular availability and metabolism of TH. To study the pathophysiology of MCT8 deficiency, we generated Mct8 knockout mice. Male mice deficient in Mct8 (Mct8( /y)) replicate the thyroid abnormalities observed in affected men. TH deprivation and replacement with L-T3 showed that suppression of TSH required higher serum levels T3 in Mct8(-/y) than wild-type (WT) littermates, indicating hypothalamus and/or thyrotroph resistance to T3. Furthermore, T4 is required to maintain the high serum T3 level because the latter was not different between the two genotypes during administration of T3. Mct8(-/y) mice have 2.3-fold higher T3 content in liver associated with 6.1- and 3.1-fold increase in deiodinase 1 mRNA and enzymatic activity, respectively. The relative T3 excess in liver of Mct8( /y) mice produced a decrease in serum cholesterol (79 +/- 18 vs. 137 +/- 38 mg/dl in WT) and an increase in alkaline phosphatase (107 +/- 23 vs. 58 +/- 3 U/liter in WT) levels. In contrast, T3 content in cerebrum was 1.8-fold lower in Mct8( /y) mice, associated with a 1.6- and 10.6-fold increase in D2 mRNA and enzymatic activity, respectively, as previously observed in TH-deprived WT mice. We conclude that cell-specific differences in intracellular TH content due to differences in contribution of the various TH transporters are responsible for the unusual clinical presentation of this defect, in contrast to TH deficiency. PMID- 16709609 TI - Derivatives of Z-bisdehydrodoisynolic acid provide a new description of the binding-activity paradox and selective estrogen receptor modulator activity. AB - Z-Bisdehydrodoisynolic acid [(+/-)-Z-BDDA], an estrogenic carboxylic acid, is highly active in vivo yet binds poorly to estrogen receptors (ERs). Studies of Z BDDA and its enantiomers demonstrate therapeutic potential as selective ER modulators; however, the activity vs. binding paradox has remained. One possible explanation is that the carboxylic acid group of Z-BDDA may be modified in vivo to an ester or amide. Synthesis of these derivatives showed the relative binding affinity (RBA) of the methyl ester for ERalpha and ERbeta was increased approximately 14- and 20-fold, respectively, relative to the parent compound. Yet, this increased affinity did not result in increased reporter gene expression. In contrast, the amide showed an unexpected approximately 4-fold decrease in RBA to both ERs compared with the parent. The relationship among the RBAs of ester, acid, and amide is consistent with their predicted polarity, suggesting the carboxylic acid, and not the carboxylate of BDDA, binds to ERs. Studies at pH 6.5, 7.4, and 8.0 were consistent with a simple acid-base equilibrium model, with BDDA binding as the undissociated acid and with affinity equal to or exceeding that of estradiol, consistent with high in vivo potency. Furthermore, the alcohol BDD-OH also demonstrated high affinity and increased activity in gene expression assays. In addition to suggesting a resolution to the decades-old binding/activity paradox, these studies may provide a direction for definitive in vivo metabolic and pharmacokinetic studies and provide additional insight into the chemical and metabolic determinants of BBDA's unique tissue selectivity and selective ER modulator activities. PMID- 16709610 TI - SIM1 overexpression partially rescues agouti yellow and diet-induced obesity by normalizing food intake. AB - Single-minded 1 (SIM1) mutations are associated with obesity in mice and humans. Haploinsufficiency of mouse Sim1 causes hyperphagic obesity with increased linear growth and enhanced sensitivity to a high-fat diet, a phenotype similar to that of agouti yellow and melanocortin 4 receptor knockout mice. To investigate the effects of increased Sim1 dosage, we generated transgenic mice that overexpress human SIM1 and examined their phenotype. Compared with wild-type mice, SIM1 transgenic mice had no obvious phenotype on a low-fat chow diet but were resistant to diet-induced obesity on a high-fat diet due to reduced food intake with no change in energy expenditure. The SIM1 transgene also completely rescued the hyperphagia and partially rescued the obesity of agouti yellow mice, in which melanocortin signaling is abrogated. Our results indicate that the melanocortin 4 receptor signals through Sim1 or its transcriptional targets in controlling food intake but not energy expenditure. PMID- 16709611 TI - Klotho protein promotes adipocyte differentiation. AB - Mice with homozygous disruption of the klotho exhibit multiple age-related disorders and have barely detectable amounts of white adipose tissue. Although klotho expression in cultured adipocytes has been reported, little is known about its function in adipocytes. In the present study, we investigated the role of klotho on adipocyte differentiation. Adipocyte differentiation was induced by incubation of confluent 3T3-L1 cells with insulin, dexamethasone, and 1-methyl-3 isobutyl-xanthin. Klotho-siRNA and expression vector were produced for klotho suppression and overexpression, respectively. Klotho protein was purified for determination of the hormonal effect of klotho. Klotho mRNA and protein expression increased up to the 3rd d of differentiation. A peroxisome proliferator-activated receptor-gamma agonist increased klotho expression during the early period of adipocyte differentiation. The mRNA expression of adipocyte differentiation markers, such as CCAAT/enhancer-binding protein (C/EBP)alpha, C/EBPbeta, C/EBPdelta, peroxisome proliferator-activated receptor-gamma, and fatty acid binding protein 4, was decreased by klotho suppression, and increased 1.9- to 3.8-fold by klotho overexpression. The results of Oil Red O staining also suggested that klotho overexpression promoted adipocyte differentiation. Klotho protein stimulation resulted in a 2.4- to 4.6-fold increase in mRNA expression of differentiation markers compared with control, and the time course depended on adipocyte induction status. Western blot analysis showed that protein levels of C/EBPalpha and C/EBPdelta were increased by Klotho protein stimulation. These results suggest that klotho works as a hormonal factor to promote adipocyte differentiation in the early days, during the period of transient proliferation in the differentiation process, and that klotho may play an essential role in adipocyte differentiation. PMID- 16709612 TI - Molecular cloning, tissue distribution, and ontogenic thyroidal expression of the chicken thyrotropin receptor. AB - TSH and the interaction with its receptor (TSHR) in the thyroid gland play a crucial role in the pituitary-thyroid axis of all vertebrates. Released upon stimulation by TSH, thyroid hormones influence numerous processes in the body and are extremely important during the last week of chicken embryonic development. In this study, we have cloned and functionally characterized the chicken TSHR (cTSHR), which was found to be a G protein-coupled receptor consisting of 10 exons. Besides the full-length cDNA, we detected two splice variants lacking either exon 3, or exons 2 and 3, both part of the extracellular domain of the receptor. Bovine TSH increased intracellular cAMP levels in HEK-239 cells transiently expressing the full-length cTSHR (EC50 = 1.43 nm). In situ hybridization showed the expression of cTSHR mRNA in the thyroidal follicular cells. cTSHR mRNA expression, as determined by real-time PCR, was also found in several other tissues such as brain, pituitary, pineal gland, and retina, suggesting that the TSH-TSHR interaction is not only important in regulating thyroid function. TSHR mRNA expression in the thyroid gland did not change significantly during the last week of embryonic development, which suggests that an increased thyroidal sensitivity is not part of the cause of the concomitant increasing T4 levels. PMID- 16709613 TI - Interleukin 11 signaling components signal transducer and activator of transcription 3 (STAT3) and suppressor of cytokine signaling 3 (SOCS3) regulate human endometrial stromal cell differentiation. AB - The differentiation of endometrial stromal cells into decidual cells (decidualization) is critical for embryo implantation, but the mechanisms remain poorly defined. Numerous paracrine agents including IL-11 promote human endometrial stromal cell (HESC) decidualization. IL-11 signaling is transduced by the signal transducers and activators of transcription (STAT) proteins. Suppressors of cytokine signaling (SOCS) proteins are stimulated in response to cytokine-inducible STAT phosphorylation, acting in a negative-feedback mechanism to hinder cytokine receptor activity. This study examined the role of IL-11 signal transduction components in HESC decidualization in an ex vivo model. Cells were induced to differentiate with estrogen plus medroxyprogesterone acetate (E+P) or cAMP (assessed by prolactin secretion) and resulted in increased STAT3 and SOCS3. E+P maximally stimulated STAT3, whereas cAMP maximally stimulated SOCS3 during decidualization, suggesting E+P and cAMP differentially regulated the signaling components. IL-11 stimulated the phosphorylation (p) of STAT3 and SOCS3 mRNA and protein. Antiprogestin (onapristone) added to decidualizing cells attenuated STAT3 protein but increased SOCS3 mRNA and protein, suggesting regulation via both ligand-dependent and -independent progesterone-receptor pathways. SOCS3 overexpression in HESC reduced IL-11-induced pSTAT3 and retarded decidualization, indicating that SOCS3 is a critical regulator of differentiation. Immunoreactive pSTAT3 and SOCS3 were all present in decidualized stromal cells, epithelial cells, and leukocytes in human endometrium. These data support a role for IL-11 via pSTAT3 and SOCS3 in initiating and progressing decidualization. PMID- 16709614 TI - Functional expression of mouse relaxin and mouse relaxin-3 in the lung from an Ebola virus glycoprotein-pseudotyped lentivirus via tracheal delivery. AB - The peptide hormone relaxin is a known modulator of connective tissue and the extracellular matrix by virtue of its ability to regulate matrix metalloproteinases (MMPs). Relaxin knockout mice exhibit age-related pulmonary fibrosis, and delivery of recombinant human H2 relaxin ameliorates fibrotic-like conditions in the mouse lung. We investigated whether lentiviral vectors (LVs) engineering the expression of murine relaxins could induce MMP activity in the mouse lung. Mouse relaxin and mouse relaxin-3 peptides engineered by recombinant LVs were biologically active as shown by stimulation of cAMP from both THP-1 and 293T cells stably expressing relaxin receptor LGR7 and by up-regulation of MMP-2 activity from primary C57BL/6 lung cell cultures. To provide the virions with enhanced tropism for the lung, LVs were pseudotyped with the Zaire strain of the Ebola virus glycoprotein (EboZ GP) and delivered by endotracheal intubation. LVs engineering luciferase pseudotyped with EboZ GP, but not with vesicular stomatitis virus glycoprotein resulted in successful LV transduction and transgene expression in C57BL/6 mouse lung by as early as d 4. Mice treated via tracheal delivery with EboZ GP pseudotyped LVs that engineered expression of mouse relaxins exhibited increased MMP-2 and MMP-9 activity in lung tissue up until the end of our study at d 21. Taken together, this study provides proof-of- principle that relaxin gene expression targeted to the mouse lungs can result in enhanced MMP activity offering potential for alleviating disease conditions characterized by dysregulation of extracellular matrix protein accumulation. PMID- 16709616 TI - Multiple-imputation for measurement-error correction. AB - BACKGROUND: There are many methods for measurement-error correction. These methods remain rarely used despite the ubiquity of measurement error. METHODS: Treating measurement error as a missing-data problem, the authors show how multiple-imputation for measurement-error (MIME) correction can be done using SAS software and evaluate the approach with a simulation experiment. RESULTS: Based on hypothetical data from a planned cohort study of 600 children with chronic kidney disease, the estimated hazard ratio for end-stage renal disease from the complete data was 2.0 [95% confidence limits (95% CL) 1.4, 2.8] and was reduced to 1.5 (95% CL 1.1, 2.1) using a misclassified exposure of low glomerular filtration rate at study entry (sensitivity of 0.9 and specificity of 0.7). The MIME correction hazard ratio was 2.0 (95% CL 1.2, 3.3), the regression calibration (RC) hazard ratio was 2.0 (95% CL 1.1, 3.7), and restriction to a 25% validation substudy yielded a hazard ratio of 2.0 (95% CL 1.0, 3.7). Based on Monte Carlo simulations across eight scenarios, MIME was approximately unbiased, had approximately correct coverage, and was sometimes more powerful than misclassified or RC analyses. Using root mean squared error as a criterion, the MIME bias correction is sometimes outweighed by added imprecision. CONCLUSION: The choice between MIME and RC depends on performance, ease, and objectives. The usefulness of MIME correction in specific applications will depend upon the sample size or the proportion validated. MIME correction may be valuable in interpreting imperfectly measured epidemiological data. PMID- 16709615 TI - Liver-specific overexpression of the insulin-like growth factor-I enhances somatic growth and partially prevents the effects of growth hormone deficiency. AB - The precise role of circulating IGF-I in somatic growth under normal and GH deficient conditions remains unclear. To define the contribution of circulating IGF-I to the endocrine regulation of somatic growth and the GH/IGF-I axis, we constructed a transgene with the transthyretin (TTR) enhancer/promoter and the mouse IGF-I cDNA and generated TTR-IGF-I transgenic mice. The transgene was exclusively expressed in the liver, which resulted in a 50-60% increase in serum IGF-I, a decrease in serum GH, and an improved tolerance to glucose challenge. The body weight and lean mass of TTR-IGF-I mice were heavier compared with wild type (WT) mice. The increase in lean mass was a result of increase in both number and thickness of skeletal muscle fibers. The femur, tibia, and body lengths of TTR-IGF-I mice also were longer. In WT mice, the GH antagonist pegvisomant (Peg) suppressed the transcription of endogenous IGF-I and acid-labile subunit (ALS) genes with no effect on IGF-binding protein 3 (IGFBP-3) mRNA. Consequently, Peg induced GH deficiency in WT mice severely reduced ALS, IGF-I, and IGFBP-3 in the circulation and caused a severe growth deficit. In TTR-IGF-I mice, Peg reduced the mRNA of the endogenous IGF-I gene with no effect on the TTR-IGF-I transgene expression, leading to a blunted decrease in serum IGF-I levels. Interestingly, IGFBP-3 mRNA was elevated and circulating IGFBP-3 was less reduced in Peg-treated TTR-IGF-I mice. Peg-treated TTR-IGF-I mice also exhibited heavier body weight and longer body length than Peg-treated WT mice. Therefore, liver-expressed IGF-I can stimulate IGFBP-3 mRNA expression and stabilize IGFBP-3 under GH deficiency, leading to a better maintenance of IGF-I levels in the circulation. Higher circulating levels of IGF-I can stimulate somatic growth and lean mass and improve glucose tolerance. PMID- 16709617 TI - Causes underlying the birth-cohort phenomenon of peptic ulcer: analysis of mortality data 1911-2000, England and Wales. AB - BACKGROUND: Since humans have been infected with Helicobacter pylori for millennia, it has remained an enigma why the occurrence of gastric and duodenal ulcer rose suddenly during 19th century. The study aim is to present a mathematical model of H. pylori epidemiology that explains the peculiar long-term trends of ulcer disease. METHODS: Gastric and duodenal ulcer mortality data from England and Wales between 1911 and 2000 were used to validate a model based on two simple and straightforward assumptions about H. pylori infection. First, the infection rate fell in the general population between 1,800 and 2,000. Second, gastric ulcer was caused by H. pylori infection contracted between the ages 5 and 15 and duodenal ulcer was caused by H. pylori infection contracted after the age of 15. As the infection receded in the general population, the two fractions of subjects who became infected between the ages 5 and 15 or after the age of 15 increased among consecutive birth cohorts. RESULTS: The analysis of the actual long-term mortality from gastric and duodenal ulcer indicates an underlying birth cohort pattern. These birth-cohort patterns of gastric and duodenal ulcer could be simulated by the interaction of two opposing time trends, namely a declining infection rate and a rising fraction of individuals acquiring their infection at increasingly older ages. The superimposition of a declining and a rising trend resulted in a bell-shaped curve of ulcer occurrence affecting consecutive birth cohorts born between 1830 and 1970. Similar to the real data, the modelled cohort pattern of gastric ulcer preceded that of duodenal ulcer by 20 years. CONCLUSION: The birth-cohort phenomenon of ulcer disease can be explained by a receding H. pylori infection accompanied by a simultaneous shift in its age of acquisition. PMID- 16709618 TI - One for every 113 inhabitants: cigarette vending machines in Germany. PMID- 16709619 TI - Trends and disparities in socioeconomic and behavioural characteristics, life expectancy, and cause-specific mortality of native-born and foreign-born populations in the United States, 1979-2003. AB - BACKGROUND: Immigrants are a growing segment of the US population. In 2003, there were 33.5 million immigrants, accounting for 12% of the total US population. Despite a rapid increase in their numbers, little information exists as to how immigrants' health and mortality profile has changed over time. In this study, we analysed trends in social and behavioural characteristics, life expectancy, and mortality patterns of immigrants and the US-born from 1979 to 2003. METHODS: We used national mortality and census data (1979-2003) and 1993 and 2003 National Health Interview Surveys to examine nativity differentials over time in health and social characteristics. Life tables, age-adjusted death rates, and logistic regression were used to examine nativity differentials. RESULTS: During 1979-81, immigrants had 2.3 years longer life expectancy than the US-born (76.2 vs 73.9 years). The difference increased to 3.4 years in 1999-2001 (80.0 vs 76.6 years). Nativity differentials in mortality increased over time for major cancers, cardiovascular diseases, diabetes, respiratory diseases, unintentional injuries, and suicide, with immigrants experiencing generally lower mortality than the US born in each period. Specifically, in 1999-2001, immigrants had at least 30% lower mortality from lung and oesophageal cancer, COPD, suicide, and HIV/AIDS, but at least 50% higher mortality from stomach and liver cancer than the US-born. Nativity differentials in mortality, health, and behavioural characteristics varied substantially by ethnicity. CONCLUSIONS: Growing ethnic heterogeneity of the immigrant population, and its migration selectivity and continuing advantages in behavioural characteristics may partly explain the overall widening health gaps between immigrants and the US-born. PMID- 16709620 TI - Work and leisure time physical activity assessed using a simple, pragmatic, validated questionnaire and incident cardiovascular disease and all-cause mortality in men and women: The European Prospective Investigation into Cancer in Norfolk prospective population study. AB - BACKGROUND: The health benefits of physical activity are well established, but the overall amount of physical activity associated with cardiovascular and other health outcomes, and whether the relationships are similar in men and women and at different ages is still debated. This may be partly related to different methods for assessing physical activity. Most studies have focused on leisure time physical activity. METHODS: We examined the prospective relationship between usual physical activity, taking into account both leisure and work activity, using a simple, pragmatic, four-point rating scale validated against heart rate monitoring, and cardiovascular disease incidence and total mortality after an average 8 years follow-up in 22,191 community living men and women aged 45-79 years with no known cardiovascular disease or cancer at baseline. RESULTS: The relative risks (95% confidence interval) for all-cause mortality (1,553 deaths) for men and women who were moderately inactive, moderately active, and active compared with those who were inactive were 0.83 (0.73-0.95), 0.68 (0.58-0.80), and 0.68 (0.57-0.81), respectively, after adjusting for age, sex, systolic blood pressure, blood cholesterol, cigarette smoking, alcohol intake, diabetes, body mass index, and social class. The relationships were also consistent for cardiovascular disease incidence (3,079 events), in subgroups stratified by age, sex, body mass index, smoking status and social class, and after excluding deaths in the first 2 years. The combined scale was more consistently associated with mortality than the individual work and leisure time components separately. CONCLUSIONS: When both work and leisure time physical activity patterns are taken into account, using a simple, pragmatic, validated questionnaire feasible for use in clinical and public health practice, even very moderate levels of usual physical activity are associated with significantly reduced risk of mortality and cardiovascular disease in men and women in the general population and potential population attributable impact of 14% for inactive compared with active levels. These findings may encourage efforts to increase physical activity levels not only in leisure time but also in usual daily working life. PMID- 16709621 TI - Diagnosing inborn errors of lipid metabolism with proton nuclear magnetic resonance spectroscopy. AB - BACKGROUND: Many severe diseases are caused by defects in lipid metabolism. As a result, patients often accumulate unusual lipids in their blood and tissues, and proper identification of these lipids is essential for correct diagnosis. In this study, we investigated the potential use of proton nuclear magnetic resonance (1H NMR) spectroscopy to simultaneously identify and quantify (un)usual lipids present in the blood of patients with different inborn errors of lipid metabolism. METHODS: We extracted blood plasma or serum lipids in chloroform methanol (2:1 by volume). After addition of the nonvolatile chemical shift and concentration reference compound octamethylcyclotetrasiloxane, we performed 1H NMR measurements on a 500-MHz spectrometer. Assignments were based on the literature, computer simulations, and reference spectra of relevant authentic standards. RESULTS: Spectra of normal plasma samples allowed the identification of 9 lipid species. We found good correlation between conventional methods and 1H NMR for cholesterol and triglyceride concentrations. We also investigated 4 inborn errors of lipid metabolism (3 in sterol metabolism and 1 in fatty acid metabolism). NMR analysis led to a correct diagnosis for all 4 diseases, whereas the concentration of the diagnostic metabolite could be determined for 3. CONCLUSIONS: 1H-NMR spectroscopy of blood plasma or serum lipid extracts can be used to accurately identify and quantify lipids. The method can also identify unusual lipids in the blood of patients with inborn errors of lipid metabolism. This technique may therefore be applicable in clinical diagnosis and follow-up. PMID- 16709622 TI - High lipoprotein-associated phospholipase A2 is a risk factor for recurrent coronary events in postinfarction patients. AB - BACKGROUND: Recent studies demonstrate that lipoprotein-associated phospholipase A2 (Lp-PLA2) is a risk factor for cardiovascular disease presumably deriving from generation of proinflammatory and proatherogenic species through its hydrolytic activity on lipoprotein-associated phospholipids. The goal of this study was to assess the relationship of Lp-PLA2 with a set of thrombogenic, lipid, inflammatory, and metabolic blood markers and to determine whether plasma Lp-PLA2 is a risk factor for recurrent coronary events in postinfarction patients. METHODS: Factor analysis on the set of blood markers and Lp-PLA2 was performed for 766 patients of the Thrombogenic Factors and Recurrent Coronary Events (THROMBO) postinfarction study. Recurrent coronary event risk was assessed as a function of blood marker concentrations and Lp-PLA2 by Cox proportional hazards multivariable regression adjusted for significant clinical covariates. RESULTS: Factor analysis revealed that Lp-PLA2 was associated with one factor dominated by cholesterol and apolipoprotein B and another factor dominated by HDL-cholesterol and triglycerides, with little association with an inflammatory factor dominated by C-reactive protein. Multivariable analysis demonstrated as significant and independent predictors of risk of secondary coronary events only apolipoprotein B in a model without Lp-PLA2 (hazard ratio, 1.66; 95% confidence interval, 1.14 2.40) and only Lp-PLA2 in a model with Lp-PLA2 included [1.90 (1.31-2.75)]. CONCLUSIONS: Lp-PLA2 is a significant and independent predictor of risk for recurrent coronary events in postinfarction patients, and Lp-PLA2 is related to both hypercholesterolemia and high triglyceride-low HDL dyslipidemia in this study population. PMID- 16709623 TI - Nanodiagnostics: a new frontier for clinical laboratory medicine. AB - BACKGROUND: The use of nanotechnologies for diagnostic applications shows great promise to meet the rigorous demands of the clinical laboratory for sensitivity and cost-effectiveness. New nanodiagnostic tools include quantum dots (QDs), gold nanoparticles, and cantilevers. QDs, which are the most promising nanostructures for diagnostic applications, are semiconductor nanocrystals characterized by high photostability, single-wavelength excitation, and size-tunable emission. QDs and magnetic nanoparticles can be used for barcoding of specific analytes. Gold and magnetic nanoparticles are key components of the bio-barcode assay, which has been proposed as a future alternative to the PCR. METHODS: We examined articles published over the past 10 years investigating the use of QDs, gold nanoparticles, cantilevers, and other nanotechnologies in promising diagnostic applications. RESULTS: Several nanodiagnostic assays have been developed, including a QD-based assay capable of detecting biotinylated prostate-specific antigen (PSA) at 0.38 ng/L, a bio-barcode assay capable of detecting 30 amol/L PSA in a 10-microL sample, and another able to detect 50 molecules of the Alzheimer marker amyloid beta-derived diffusible ligand in 10 microL of cerebrospinal fluid. CONCLUSIONS: Nanodiagnostics promise increased sensitivity, multiplexing capabilities, and reduced cost for many diagnostic applications as well as intracellular imaging. Further work is needed to fully optimize these diagnostic nanotechnologies for clinical laboratory setting and to address the potential health and environmental risks related to QDs. PMID- 16709624 TI - Gc-globulin: roles in response to injury. AB - BACKGROUND: Gc-globulin (vitamin D-binding protein) appears to have important functions in addition to its role as a carrier of vitamin D. APPROACH: We reviewed recent studies focusing on the pathophysiologic functions and clinical significance of Gc-globulin. RESULTS: Serum concentrations of Gc-globulin, as determined by immunoassay techniques, are decreased in severe injury. The extent of the decrease may have prognostic significance for patient outcomes. Clinical studies and animal models have shown that Gc-globulin has an important role in the clearance of procoagulant actin from the circulation after its release during cell necrosis and tissue injury. Gc-globulin has other potential roles in responses to acute tissue injury through conversion to a macrophage-activating factor, neutrophil chemotactic activity, and enhancement of C5a-mediated signaling. CONCLUSION: Considering the important physiologic roles of Gc-globulin in responses to tissue injury, such as clearance of actin, measurement of Gc globulin may have value in directing the care of patients in many clinical disorders. PMID- 16709625 TI - Liquid chromatography-tandem mass spectrometry for detection of low concentrations of 21 benzodiazepines, metabolites, and analogs in urine: method with forensic applications. AB - BACKGROUND: Commonly used methods for detecting benzodiazepines (BZPs) and BZP like substances, such as zolpidem and zopiclone, may not detect low concentrations of these drugs. We developed a liquid chromatographic-tandem mass spectrometric method for identifying these drugs and their relevant metabolites. METHODS: We extracted BZPs from urine by solid-phase extraction with a mixed-mode phase (OASIS HLB cartridges). Chromatographic separation was performed with a Waters XTerra MS C18 [150 x 2.1 mm (i.d.); bead size, 5 microm] reversed-phase column with deuterated analogs of the analytes as internal standards (IS). Detection was performed with a triple-quadruple mass spectrometer that monitored 2 specific transitions per compound in the electrospray, positive-ion selected reaction monitoring mode. We tested this technique on urine samples from 12 healthy volunteers and 1 forensic sample obtained in a case of alleged drug facilitated sexual assault. RESULTS: Chromatographic separation was achieved within 18 min. The linear dynamic ranges extended from 0.02 or 0.1 microg/L (depending on the drug or metabolite) to 50 microg/L. Extraction recovery (range) was 77%-110%. Limits of detection were < or = 0.05 microg/L. No ion suppression was seen except for alprazolam, for which baseline decreased by almost 20%. In the forensic urine sample, the method detected alprazolam (3.5 microg/L) and its characteristic metabolite, alpha-hydroxyalprazolam (0.17 microg/L). CONCLUSION: This method measured low concentrations of BZPs and BZP-like substances and might be useful for analyses of urine in suspected drug-facilitated sexual assault cases. PMID- 16709626 TI - Sensitive and quantitative measurement of gene expression directly from a small amount of whole blood. AB - BACKGROUND: Accurate and precise quantification of mRNA in whole blood is made difficult by gene expression changes during blood processing, and by variations and biases introduced by sample preparations. We sought to develop a quantitative whole-blood mRNA assay that eliminates blood purification, RNA isolation, reverse transcription, and target amplification while providing high-quality data in an easy assay format. METHODS: We performed single- and multiplex gene expression analysis with multiple hybridization probes to capture mRNA directly from blood lysate and used branched DNA to amplify the signal. The 96-well plate singleplex assay uses chemiluminescence detection, and the multiplex assay combines Luminex encoded beads with fluorescent detection. RESULTS: The single- and multiplex assays could quantitatively measure as few as 6000 and 24,000 mRNA target molecules (0.01 and 0.04 amoles), respectively, in up to 25 microL of whole blood. Both formats had CVs < 10% and dynamic ranges of 3-4 logs. Assay sensitivities allowed quantitative measurement of gene expression in the minority of cells in whole blood. The signals from whole-blood lysate correlated well with signals from purified RNA of the same sample, and absolute mRNA quantification results from the assay were similar to those obtained by quantitative reverse transcription-PCR. Both single- and multiplex assay formats were compatible with common anticoagulants and PAXgene-treated samples; however, PAXgene preparations induced expression of known antiapoptotic genes in whole blood. CONCLUSIONS: Both the singleplex and the multiplex branched DNA assays can quantitatively measure mRNA expression directly from small volumes of whole blood. The assay offers an alternative to current technologies that depend on RNA isolation and is amenable to high-throughput gene expression analysis of whole blood. PMID- 16709627 TI - AMP-activated protein kinase: function and dysfunction in health and disease. PMID- 16709628 TI - Queer channels in hippocampal basket cells: h-current without sag. PMID- 16709630 TI - Single-channel analysis of a point mutation of a conserved serine residue in the S2 ligand-binding domain of the NR2A NMDA receptor subunit. AB - We have examined the function of a conserved serine residue (Ser670) in the S2 ligand-binding region of the NR2A N-methyl-d-aspartate (NMDA) receptor subunit, using recombinant NR1/NR2A receptors expressed in Xenopus laevis oocytes. Mutation of Ser670 to glycine (S670G) in NR2A reduced the potency of glutamate by 124-fold. Single-channel conductance and the duration of apparent open periods of NR2A(S670G) receptor mutants were, however, indistinguishable from wild-type NMDA receptors. NR1/NR2A(S670G) shut-time distributions were best described by a mixture of six exponential components, and the four shortest shut intervals of each distribution were considered to occur within a channel activation (burst). Bursts of single-channel openings were fitted with a mixture of four exponential components. The longest two components carried the majority of the charge transfer and had mean durations of 9.6 +/- 0.5 and 29.6 +/- 1.5 ms. The overall channel open probability during a burst was high (mean, 0.83 +/- 0.06). Consistent with a shortening of NMDA receptor-channel burst lengths was the observation of an increased deactivation rate of macroscopic currents evoked by brief applications of glutamate to outside-out membrane patches. Correlations between shut times and adjacent open times were observed in all data records. Noticeably, shorter than average openings tended to occur next to long closed periods, whereas longer than average openings tended to occur next to short closings. Our single-channel data, together with modelling using a kinetic scheme to describe channel activations, support our hypothesis that the S670G point mutation reduces the dwell time of glutamate in its binding site. PMID- 16709629 TI - AMPK integrates nutrient and hormonal signals to regulate food intake and energy balance through effects in the hypothalamus and peripheral tissues. AB - The evolutionarily conserved serine/threonine kinase, AMP-activated protein kinase (AMPK), functions as a cellular fuel gauge that regulates metabolic pathways in glucose and fatty acid metabolism and protein synthesis. Recent data strongly implicate the AMPK-acetyl CoA carboxylase (ACC)-malonyl CoA pathway in the hypothalamus in the regulation of food intake, body weight and hepatic glucose production. Furthermore, data indicate that AMPK is a mediator of the effects of adipocyte-derived and gut-derived hormones and peptides on fatty acid oxidation and glucose uptake in peripheral tissues. Studies are now elucidating the potential role of kinases upstream of AMPK in these metabolic effects. In addition, recently, several novel downstream effectors of AMPK have been identified. The AMPK pathway in the hypothalamus and peripheral tissues coordinately integrates inputs from multiple hormones, peptides and nutrients to maintain energy homeostasis. PMID- 16709631 TI - Temperature change does not affect force between regulated actin filaments and heavy meromyosin in single-molecule experiments. AB - The temperature dependence of sliding velocity, force and the number of cross bridges was studied on regulated actin filaments (reconstituted thin filaments) when they were placed on heavy meromyosin (HMM) attached to a glass surface. The regulated actin filaments were used because our previous study on muscle fibres demonstrated that the temperature effect was much reduced in the absence of regulatory proteins. A fluorescently labelled thin filament was attached to the gelsolin-coated surface of a polystyrene bead. The bead was trapped by optical tweezers, and HMM-thin filament interaction was performed at 20-35 degrees C to study the temperature dependence of force at the single-molecule level. Our experiments showed that there was a small increase in force with temperature (Q10 = 1.43) and sliding velocity (Q10 = 1.46). The small increase in force was correlated with the small increase in the number of cross-bridges (Q10 = 1.49), and when force was divided by the number of cross-bridges, the result did not depend on the temperature (Q(10) = 1.03). These results demonstrate that the force each cross-bridge generates is fixed and independent of temperature. Our additional experiments demonstrate that tropomyosin (Tm) in the presence of troponin (Tn) and Ca2+ enhances both force and velocity, and a truncated mutant, Delta23Tm, diminishes force and velocity. These results are consistent with the hypothesis that Tm in the presence of Tn and Ca2+ exerts a positive allosteric effect on actin to make actomyosin linkage more secure so that larger forces can be generated. PMID- 16709632 TI - Functions of AMP-activated protein kinase in adipose tissue. AB - AMP-activated protein kinase (AMPK) is involved in cellular energy homeostasis. Its functions have been extensively studied in muscles and liver. AMPK stimulates pathways which increase energy production (glucose transport, fatty acid oxidation) and switches off pathways which consume energy (lipogenesis, protein synthesis, gluconeogenesis). This has led to the concept that AMPK has an interesting pharmaceutical potential in situations of insulin resistance and it is indeed the target of existing drugs and hormones which improve insulin sensitivity. Adipose tissue is a key player in energy metabolism through the release of substrates and hormones involved in metabolism and insulin sensitivity. Activation of AMPK in adipose tissue can be achieved through situations such as fasting and exercise. Leptin and adiponectin as well as hypoglycaemic drugs are activators of adipose tissue AMPK. This activation probably involves changes in the AMP/ATP ratio and the upstream kinase LKB1. When activated, AMPK limits fatty acid efflux from adipocytes and favours local fatty acid oxidation. Since fatty acids have a key role in insulin resistance, especially in muscles, activating AMPK in adipose tissue might be found to be beneficial in insulin-resistant states, particularly as AMPK activation also reduces cytokine secretion in adipocytes. PMID- 16709633 TI - BDNF increases release probability and the size of a rapidly recycling vesicle pool within rat hippocampal excitatory synapses. AB - Exerting its actions pre-, post- and peri-synaptically, brain-derived neurotrophic factor (BDNF) is one of the most potent modulators of hippocampal synaptic function. Here, we examined the effects of BDNF on a rapidly recycling pool (RRP) of vesicles within excitatory synapses. First, we estimated vesicular release in hippocampal cultures by performing FM4-64 imaging in terminals impinging on enhanced green fluorescent protein (eGFP)-labelled dendritic spines a hallmark of excitatory synapses. Consistent with a modulation of the RRP, BDNF increased the evoked destaining rate of FM4-64 only during the initial phase of field stimulation. Multiphoton microscopy in acute hippocampal slices confirmed these observations by selectively imaging the RRP, which was loaded with FM1-43 by hyperosmotic shock. Slices exposed to BDNF showed an increase in the evoked and spontaneous rates of FM1-43 destaining from terminals in CA1 stratum radiatum, mostly representing excitatory terminals of Schaffer collaterals. Variance-mean analysis of evoked EPSCs in CA1 pyramidal neurons further confirmed that release probability is increased in BDNF-treated slices, without changes in the number of independent release sites or average postsynaptic quantal amplitude. Because BDNF was absent during dye loading, imaging, destaining and whole-cell recordings, these results demonstrate that BDNF induces a long-lasting enhancement in the probability of transmitter release at hippocampal excitatory synapses by modulating the RRP. Since the endogenous BDNF scavenger TrkB-IgG prevented the enhancement of FM1-43 destaining rate caused by induction of long term potentiation in acute hippocampal slices, the modulation of a rapidly recycling vesicle pool may underlie the role of BDNF in hippocampal long-term synaptic plasticity. PMID- 16709634 TI - Delayed synchronization of activity in cortex and subthalamic nucleus following cortical stimulation in the rat. AB - Oscillations may play a role in the functional organization of cortico-basal ganglia-thalamocortical circuits, and it is important to understand their underlying mechanisms. The cortex often drives basal ganglia (BG) activity, and particularly, oscillatory activity in the subthalamic nucleus (STN). However, the STN may also indirectly influence cortex. The aim of this study was to characterize the delayed (>200 ms) responses of STN neurons to synchronized cortical inputs, focusing on their relationship with oscillatory cortical activity. We recorded the short-latency and delayed responses of STN units and frontal electrocorticogram (ECoG) to cortical stimulation in anaesthetized rats. Similar to previous studies, stimulation of ipsilateral frontal cortex, but not temporal cortex, evoked a short-latency triphasic response, followed by a sustained reduction or pause in firing, in rostral STN units. Caudal STN units did not show the short-latency triphasic response but often displayed a prolonged firing reduction. Oscillations in STN unit activity and ECoG were common after this sustained firing reduction, particularly between 200 and 600 ms after frontal cortical stimulation. These delayed oscillations were significantly coherent in a broad frequency band of 5-30 Hz. Coherence with ECoG at 5-15 Hz was observed throughout STN, though coherence at 15-30 Hz was largely restricted to rostral STN. Furthermore, oscillatory responses at 5-30 Hz in rostral STN predominantly led those in cortex (mean latency of 29 ms) after frontal cortical stimulation. These findings suggest that STN neurons responding to corticosubthalamic inputs may provide a delayed input to cortex, via BG output nuclei, and thence, thalamocortical pathways. PMID- 16709635 TI - 'Simplification' of responses of complex cells in cat striate cortex: suppressive surrounds and 'feedback' inactivation. AB - In mammalian striate cortex (V1), two distinct functional classes of neurones, the so-called simple and complex cells, are routinely distinguished. They can be quantitatively differentiated from each other on the basis of the ratio between the phase-variant (F1) component and the mean firing rate (F0) of spike responses to luminance-modulated sinusoidal gratings (simple, F1/F0 > 1; complex, F1/F0 < 1). We investigated how recurrent cortico-cortical connections affect the spatial phase-variance of responses of V1 cells in the cat. F1/F0 ratios of the responses to optimally oriented drifting sine-wave gratings covering the classical receptive field (CRF) of single V1 cells were compared to those of: (1) responses to gratings covering the CRFs combined with gratings of different orientations presented to the 'silent' surrounds; and (2) responses to CRF stimulation during reversible inactivation of postero-temporal visual (PTV) cortex. For complex cells, the relative strength of the silent surround suppression on CRF-driven responses was positively correlated with the extent of increases in F1/F0 ratios. Inactivation of PTV cortex increased F1/F0 ratios of CRF-driven responses of complex cells only. Overall, activation of suppressive surrounds or inactivation of PTV 'converted' substantial proportions (50 and 30%, respectively) of complex cells into simple-like cells (F1/F0 > 1). Thus, the simple-complex distinction depends, at least partly, on information coming from the silent surrounds and/or feedback from 'higher-order' cortices. These results support the idea that simple and complex cells belong to the same basic cortical circuit and the spatial phase variance of their responses depends on the relative strength of different synaptic inputs. PMID- 16709636 TI - Hypersensitivity of pulmonary chemosensitive neurons induced by activation of protease-activated receptor-2 in rats. AB - This study was carried out to determine the effect of protease-activated receptor 2 (PAR2) activation on the pulmonary chemoreflex responses and on the sensitivity of isolated rat vagal pulmonary chemosensitive neurons. In anaesthetized, spontaneously breathing rats, intratracheal instillation of trypsin (0.8 mg ml( 1), 0.1 ml), an endogenous agonist of PAR2, significantly amplified the capsaicin induced pulmonary chemoreflex responses. The enhanced responses were completely abolished by perineural capsaicin treatment of both cervical vagi, suggesting the involvement of pulmonary C-fibre afferents. In patch-clamp recording experiments, pretreatment with trypsin (0.1 microM, 2 min) potentiated the capsaicin-induced whole-cell inward current in isolated pulmonary sensory neurons. The potentiating effect of trypsin was mimicked by PAR2-activating peptide (PAR2-AP) in a concentration-dependent manner. PAR2-AP pretreatment (100 microM, 2 min) also markedly enhanced the acid-evoked inward currents in these sensory neurons. Furthermore, the sensitizing effect of PAR2 was completely abolished by pretreatment with either U73122 (1 microM, 4 min), a phospholipase C inhibitor, or chelerythrine (10 microM, 4 min), a protein kinase C (PKC) inhibitor. In summary, our results have demonstrated that activation of PAR2 upregulates the pulmonary chemoreflex sensitivity in vivo and the excitability of isolated pulmonary chemosensitive neurons in vitro, and this effect of PAR2 activation was mediated through the PKC-dependent transduction pathway. These results further suggest that the hypersensitivity of these neurons may play a part in the development of airway hyper-responsiveness resulting from PAR2 activation. PMID- 16709637 TI - The role of AMP-activated protein kinase in mitochondrial biogenesis. AB - While it has been known for more than 75 years that physical activity is associated with increased mitochondrial content in muscle, the molecular mechanism for this adaptive process has only recently been elucidated. This brief review examines existing studies that have identified AMPK-activated protein kinase (AMPK) and several other key regulators of mitochondrial biogenesis, including peroxisome proliferator-activated receptor-gamma coactivator-1alpha and -1beta, calcium/calmodulin-dependent protein kinase IV, and nitric oxide. In addition, the potential role of mitochondrial dysfunction in the pathogenesis of insulin resistance associated with ageing and type 2 diabetes mellitus is also discussed. PMID- 16709638 TI - Effect of repetitive hypoxic apnoeas on baroreflex function in humans. AB - Baroreflex function is impaired in patients with obstructive sleep apnoea. We tested the hypothesis that short-term exposure to repetitive hypoxic apnoeas (RHA) produces prolonged impairment in baroreflex function. Baroreflex function was determined using the modified Oxford technique in 14 subjects (26 +/- 1 years). Baroreflex sensitivity (BRS) was quantified from the R-R interval systolic blood pressure (BP; cardiovagal BRS), heart rate-systolic BP (HR BRS) and muscle sympathetic nerve activity (MSNA)-diastolic BP (sympathetic BRS) relations. RHA involved subjects performing repetitive end-expiratory apnoeas (20 s) every minute for 30 min during intermittent hypoxia to accentuate oxygen desaturation. After RHA, BP and MSNA at rest were elevated. BRS was measured approximately 7 (Post 1), approximately 30 (Post 2) and approximately 50 min (Post 3) after RHA to provide insight into the temporal pattern of responses. Cardiovagal BRS (16.8 +/- 1.3, 16.5 +/- 1.6, 17.6 +/- 2.0 and 17.4 +/- 1.5 ms mmHg(-1) for Pre, Post 1, Post 2 and Post 3, respectively), HR BRS (-1.1 +/- 0.1, -1.1 +/- 0.1, -1.3 +/- 0.1 and -1.4 +/- 0.1 beats min(-1) mmHg(-1)) and sympathetic BRS (-4.5 +/- 0.6, -4.4 +/- 0.7, -3.7 +/- 0.5 and -4.7 +/- 1.0 arbitrary units (au) beat(-1) mmHg(-1)) were unchanged by RHA. In contrast, the operating points of the baroreflexes were shifted rightward (to higher levels of BP) and upward (to higher levels of heart rate and MSNA) after RHA (P < 0.05). Time control studies performed in five additional subjects showed no change in any of the measured variables over time. Collectively, these data indicate that short-term exposure to RHA shifts ('resets') the baroreflex stimulus-response curve to higher levels of BP without influencing BRS for extended periods of time. PMID- 16709639 TI - AMP-activated protein kinase and the regulation of Ca2+ signalling in O2-sensing cells. AB - All cells respond to metabolic stress. However, a variety of specialized cells, commonly referred to as O2-sensing cells, are acutely sensitive to relatively small changes in PO2. Within a variety of organisms such O2-sensing cells have evolved as vital homeostatic mechanisms that monitor O2 supply and alter respiratory and circulatory function, as well as the capacity of the blood to transport O2. Thereby, arterial PO2 may be maintained within physiological limits. In mammals, for example, two key tissues that contribute to this process are the pulmonary arteries and the carotid bodies. Constriction of pulmonary arteries by hypoxia optimizes ventilation-perfusion matching in the lung, whilst carotid body excitation by hypoxia initiates corrective changes in breathing patterns via increased sensory afferent discharge to the brain stem. Despite extensive investigation, the precise mechanism(s) by which hypoxia mediates these responses has remained elusive. It is clear, however, that hypoxia inhibits mitochondrial function in O2-sensing cells over a range of PO2 that has no such effect on other cell types. This raised the possibility that AMP-activated protein kinase might function to couple mitochondrial oxidative phosphorylation to Ca2+ signalling mechanisms in O2-sensing cells and thereby underpin pulmonary artery constriction and carotid body excitation by hypoxia. Our recent investigations have provided significant evidence in support of this view. PMID- 16709640 TI - Different responses of rat cerebellar Purkinje cells and Golgi cells evoked by widespread convergent sensory inputs. AB - While the synaptic properties of Golgi cell-mediated inhibition of granule cells are well studied, less is known of the afferent inputs to Golgi cells so their role in information processing remains unclear. We investigated the responses of cerebellar cortical Golgi cells and Purkinje cells in Crus I and II of the posterior lobe cerebellar hemisphere to activation of peripheral afferents in vivo, using anaesthetized rats. Recordings were made from 70 Golgi cells and 76 Purkinje cells. Purkinje cells were identified by the presence of climbing fibre responses. Golgi cells were identified by both spontaneous firing pattern and response properties, and identification was confirmed using juxtacellular labelling of single neurones (n = 16). Purkinje cells in Crus II showed continuous firing at relatively high rates (25-60 Hz) and stimulation of peripheral afferents rarely evoked substantial responses. The most common response was a modest, long-latency, long-lasting increase in simple spike output. By comparison, the most common response evoked in Golgi cells by the same stimuli was a long-latency, long-lasting depression of firing, found in approximately 70% of the Golgi cells tested. The onsets of Golgi cell depressions had shorter latencies than the Purkinje cell excitations. Brief, short-latency excitations and reductions in firing were also evoked in some Golgi cells, and rarely in Purkinje cells, but in most cases long-lasting depressions were the only significant change in spike firing. Golgi cell responses could be evoked using air puff or tactile stimuli and under four different anaesthetic regimens. Long-lasting responses in both neurone types could be evoked from wide receptive fields, in many cases including distal afferents from all four limbs, as well as from trigeminal afferents. These Golgi cell responses are not consistent with the conventional feedback inhibition or 'gain control' models of Golgi cell function. They suggest instead that cerebellar cortical activity can be powerfully modulated by the general level of peripheral afferent activation from much of the body. On this basis, Golgi cells may act as a context-specific gate on transmission through the mossy fibre-granule cell pathway. PMID- 16709641 TI - Residual force enhancement in skeletal muscle. AB - Residual force enhancement has been observed consistently in skeletal muscles following active stretching. However, its underlying mechanism(s) remain elusive, and it cannot be explained readily within the framework of the cross-bridge theory. Traditionally, residual force enhancement has been attributed to the development of sarcomere length non-uniformities. However, recent evidence suggests that this might not be the case. Rather, it appears that residual force enhancement has an active and a passive component. The active component is tentatively associated with changes in the cross-bridge kinetics that might be reflected in decreased detachment rates following active muscle stretching, while the passive component possibly originates from a structural protein, such as titin, whose stiffness might be regulated by calcium. PMID- 16709642 TI - T-tubule disorganization and reduced synchrony of Ca2+ release in murine cardiomyocytes following myocardial infarction. AB - In cardiac myocytes, initiation of excitation-contraction coupling is highly localized near the T-tubule network. Myocytes with a dense T-tubule network exhibit rapid and homogeneous sarcoplasmic reticulum (SR) Ca(2+) release throughout the cell. We examined whether progressive changes in T-tubule organization and Ca(2+) release synchrony occur in a murine model of congestive heart failure (CHF). Myocardial infarction (MI) was induced by ligation of the left coronary artery, and CHF was diagnosed by echocardiography (left atrial diameter >2.0 mm). CHF mice were killed at 1 or 3 weeks following MI (1-week CHF, 3-week CHF) and cardiomyocytes were isolated from viable regions of the septum, excluding the MI border zone. Septal myocytes from SHAM-operated mice served as controls. T-tubules were visualized by confocal microscopy in cells stained with di-8-ANEPPS. SHAM cells exhibited a regular striated T-tubule pattern. However, 1 week CHF cells showed slightly disorganized T-tubule structure, and more profound disorganization occurred in 3-week CHF with irregular gaps between adjacent T tubules. Line-scan images of Ca(2+) transients (fluo-4 AM, 1 Hz) showed that regions of delayed Ca(2+) release occurred at these gaps. Three-week CHF cells exhibited an increased number of delayed release regions, and increased overall dyssynchrony of Ca(2+) release. A common pattern of Ca(2+) release in 3-week CHF was maintained between consecutive transients, and was not altered by forskolin application. Thus, progressive T-tubule disorganization during CHF promotes dyssynchrony of SR Ca(2+) release which may contribute to the slowing of SR Ca(2+) release in this condition. PMID- 16709643 TI - Role of endothelin receptor activation in secondary pulmonary hypertension in awake swine after myocardial infarction. AB - We previously observed that pulmonary hypertension secondary to myocardial infarction (MI) in swine is characterized by elevated plasma endothelin (ET) levels and pulmonary vascular resistance (PVR). Consequently, we tested the hypothesis that an increased ET-mediated vasoconstrictor influence contributes to secondary pulmonary hypertension after MI and investigated the involvement of ET(A) and ET(B) receptor subtypes. Chronically instrumented swine with (MI swine; n = 25) or without (normal swine; n = 19) MI were studied at rest and during treadmill exercise (up to 4 km h(-1)), in the absence and presence of the ET(A) antagonist EMD 122946 or the mixed ET(A)/ET(B) antagonist tezosentan. In normal swine, exercise caused a small decrease in PVR. ET(A) blockade had no effect on PVR at rest or during exercise. Conversely, ET(A)/ET(B) blockade decreased PVR but only during exercise (at 4 km h(-1), from 3.0 +/- 0.1 to 2.3 +/- 0.1 mmHg min l(-1); P 30, 60 min > fatigue), Ca(2+) uptake (0 > 30 > 60 min, fatigue), and Ca(2+) release, both phase 1 (0, 30 > 60 min, fatigue) and phase 2 (0 > 30, 60 min, fatigue; 30 min > fatigue) in Norm CHO. Exercise was without effect in altering the Hill slope (n(H)), defined as the slope of relationship between Ca(2+)-ATPase activity and Ca(2+) concentration. No differences were observed between Norm CHO and Ex+Norm CHO. Compared with Norm CHO, Lo CHO resulted in a lower (P < 0.05) Ca(2+) uptake, phase 1 Ca(2+) release (30 min), and n(H). Ex + Lo CHO resulted in a greater (P < 0.05) Ca(2+) uptake and n(H) compared with Lo CHO. The results demonstrate that Lo CHO alone can disrupt SR Ca(2+) cycling and that, with the exception of Ca(2+) release, a glycogen-depleting session of exercise before Lo CHO can reverse the effects. PMID- 16709651 TI - Preserved autonomic function in amenorrheic athletes. AB - Reproductive hormones such as estradiol and progesterone are known to influence autonomic cardiovascular regulation. The purpose of this study was to determine whether amenorrheic athletes (AA) have impaired autonomic cardiovascular regulation compared with eumenorrheic athletes (EA). Thirty-five athletes were tested: 13 AA (19 +/- 1 yr), 13 EA (21 +/- 1 yr), and 9 EA (23 +/- 1 yr) on oral contraceptives (EA-OC). Multiple indexes of autonomic cardiovascular regulation were assessed: respiratory sinus arrhythmia (RSA), cardiovagal baroreflex sensitivity (BRS) via phase IV and phase II of the Valsalva maneuver, a spontaneous index of BRS, and the heart rate and blood pressure responses to orthostatic stress (20-min 60 degrees head-up tilt). RSA was not different among the groups. There were no group differences in the spontaneous index of BRS (AA = 30 +/- 6, EA = 24 +/- 3, EA-OC = 29 +/- 5 ms/mmHg) or in phase II (AA = 8 +/- 2, EA = 7 +/- 1, EA-OC = 8 +/- 1 ms/mmHg) of the Valsalva. There was a difference in BRS during phase IV (AA = 21 +/- 3, EA = 15 +/- 1, EA-OC = 26 +/- 6 ms/mmHg; ANOVA P = 0.04). Tukey's post hoc test indicated that BRS was greater in the EA OC group compared with the EA group (P = 0.04). There were no differences in cardiovascular responses to orthostatic stress among the groups. In conclusion, AA do not display signs of impaired autonomic function and orthostatic responses compared with EA or EA-OC during the follicular phase of the menstrual cycle. PMID- 16709649 TI - Contractile properties of EDL and soleus muscles of myostatin-deficient mice. AB - Myostatin is a negative regulator of muscle mass. The impact of myostatin deficiency on the contractile properties of healthy muscles has not been determined. We hypothesized that myostatin deficiency would increase the maximum tetanic force (P(o)), but decrease the specific P(o) (sP(o)) of muscles and increase the susceptibility to contraction-induced injury. The in vitro contractile properties of extensor digitorum longus (EDL) and soleus muscles from wild-type (MSTN(+/+)), heterozygous-null (MSTN(+/-)), and homozygous-null (MSTN( /-)) adult male mice were determined. For EDL muscles, the P(o) of both MSTN(+/-) and MSTN(-/-) mice were greater than the P(o) of MSTN(+/+) mice. For soleus muscles, the P(o) of MSTN(-/-) mice was greater than that of MSTN(+/+) mice. The sP(o) of EDL muscles of MSTN(-/-) mice was less than that of MSTN(+/+) mice. For soleus muscles, however, no difference in sP(o) was observed. Following two lengthening contractions, EDL muscles from MSTN(-/-) mice had a greater force deficit than that of MSTN(+/+) or MSTN(+/-) mice, whereas no differences were observed for the force deficits of soleus muscles. Myostatin-deficient EDL muscles had less hydroxyproline, and myostatin directly increased type I collagen mRNA expression and protein content. The difference in the response of EDL and soleus muscles to myostatin may arise from differences in the levels of a myostatin receptor, activin type IIB. Compared with the soleus, the amount of activin type IIB receptor was approximately twofold greater in EDL muscles. The results support a significant role for myostatin not only in the mass of muscles but also in the contractility and the composition of the extracellular matrix of muscles. PMID- 16709652 TI - Pharyngeal critical pressure in children with mild sleep-disordered breathing. AB - There is evidence that narrowing or collapse of the pharynx can contribute to obstructive sleep-disordered breathing (SDB) in adults and children. However, studies in children have focused on those with relatively severe SDB who generally were recruited from sleep clinics. It is unclear whether children with mild SDB who primarily have hypopneas, and not frank apnea, also have more collapsible airways. We estimated airway collapsibility in 10 control subjects (9.4 +/- 0.5 yr old; 1.9 +/- 0.2 hypopneas/h) and 7 children with mild SDB (10.6 +/- 0.5 yr old; 11.5 +/- 0.1 hypopneas/h) during stable, non-rapid eye movement sleep. None of the subjects had clinically significant enlargement of the tonsils or adenoids, nor had any undergone previous tonsillectomy or adenoidectomy. Airway collapsibility was measured by brief (2-breath duration) and sudden reductions in pharyngeal pressure by connecting the breathing mask to a negative pressure source. Negative pressure applications ranging from -1 to -20 cmH(2)O were randomly applied in each subject while respiratory airflow and mask pressure were measured. Flow-pressure curves were constructed for each subject, and the x intercept gave the pressure at zero flow, the so-called critical pressure of the upper airway (Pcrit). Pcrit was significantly higher in children with SDB than in controls (-10.8 +/- 2.8 vs. -15.7 +/- 1.2 cmH(2)O; P < 0.05). There were no significant differences in the slopes of the pressure-flow relations or in baseline airflow resistance. These data support the concept that intrinsic pharyngeal collapsibility contributes to mild SDB in children. PMID- 16709653 TI - Of balance and unbalance. PMID- 16709655 TI - The Last Word: Point:Counterpoint authors respond to commentaries on "the parafacial respiratory group (pFRG)/pre-Botzinger complex (preBotC) is the primary site of respiratory rhythm generation in the mammal". PMID- 16709656 TI - Effect of aging on the recovery following contraction-induced injury in muscles of female mice. AB - By the age of 80 yr, the skeletal muscles of men and women decrease in mass and maximum force by approximately 30%. Severe contraction-induced injury may contribute to these age-related declines. One to two months after a 225 lengthening contraction protocol (LCP), muscles of young/adult male mice recovered completely, whereas those of old male mice sustained deficits of approximately 15% in mass and approximately 25% in maximum force. Although gender related differences in the early events of contraction-induced injury have been reported, the recovery phase of muscles in old female animals has not been investigated. The hypothesis tested was that 2 mo after a severe LCP to the plantar flexor muscle group, the magnitude of recovery of mass and force for old female mice is less than that for adult female mice. The LCP was administered to muscles of adult and old, female C57BL/6 mice. At 3 days, 1 mo, and 2 mo following the LCP, maximum isometric force was measured, and muscles were removed and weighed. Two months following the LCP, the muscles of adult female mice recovered mass and force. In contrast, for old female mice, even after 2 mo, muscle masses were decreased by 11% and maximum forces by 38%. We conclude that, as reported previously for old male mice, a severe contraction-induced injury to muscles of old female mice results in prolonged deficits in mass and force. PMID- 16709657 TI - Influence of body heat content on hand function during prolonged cold exposures. AB - We examined the influence of 1) prior increase [preheating (PHT)], 2) increase throughout [heating (HT)], and 3) no increase [control (Con)] of body heat content (H(b)) on neuromuscular function and manual dexterity of the hands during a 130-min exposure to -20 degrees C (coldEx). Ten volunteers randomly underwent three passive coldEx, incorporating a 10-min moderate-exercise period at the 65th min while wearing a liquid conditioning garment (LCG) and military arctic clothing. In PHT, 50 degrees C water was circulated in the LCG before coldEx until core temperature was increased by 0.5 degrees C. In HT, participants regulated the inlet LCG water temperature throughout coldEx to subjective comfort, while the LCG was not operating in Con. Thermal comfort, rectal temperature, mean skin temperature, mean finger temperature (T(fing)), change in H(b) (DeltaH(b)), rate of body heat storage, Purdue pegboard test, finger tapping, handgrip, maximum voluntary contraction, and evoked twitch force of the first dorsal interosseus muscle were recorded. Results demonstrated that, unlike in HT and PHT, thermal comfort, rectal temperature, mean skin temperature, twitch force, maximum voluntary contraction, and finger tapping declined significantly in Con. In contrast, T(fing) and Purdue pegboard test remained constant only in HT. Generalized estimating equations demonstrated that DeltaH(b) and T(fing) were associated over time with hand function, whereas no significant association was detected for rate of body heat storage. It is concluded that increasing H(b) not only throughout but also before a coldEx is effective in maintaining hand function. In addition, we found that the best indicator of hand function is DeltaH(b) followed by T(fing). PMID- 16709658 TI - Spectrum of gluten-sensitive enteropathy in first-degree relatives of patients with coeliac disease: clinical relevance of lymphocytic enteritis. AB - BACKGROUND: Limited data on a short series of patients suggest that lymphocytic enteritis (classically considered as latent coeliac disease) may produce symptoms of malabsorption, although the true prevalence of this situation is unknown. Serological markers of coeliac disease are of little diagnostic value in identifying these patients. AIMS: To evaluate the usefulness of human leucocyte antigen-DQ2 genotyping followed by duodenal biopsy for the detection of gluten sensitive enteropathy in first-degree relatives of patients with coeliac disease and to assess the clinical relevance of lymphocytic enteritis diagnosed with this screening strategy. PATIENTS AND METHODS: 221 first-degree relatives of 82 DQ2+ patients with coeliac disease were consecutively included. Duodenal biopsy (for histological examination and tissue transglutaminase antibody assay in culture supernatant) was carried out on all DQ2+ relatives. Clinical features, biochemical parameters and bone mineral density were recorded. RESULTS: 130 relatives (58.8%) were DQ2+, showing the following histological stages: 64 (49.2%) Marsh 0; 32 (24.6%) Marsh I; 1 (0.8%) Marsh II; 13 (10.0%) Marsh III; 15.4% refused the biopsy. 49 relatives showed gluten sensitive enteropathy, 46 with histological abnormalities and 3 with Marsh 0 but positive tissue transglutaminase antibody in culture supernatant. Only 17 of 221 relatives had positive serological markers. Differences in the diagnostic yield between the proposed strategy and serology were significant (22.2% v 7.2%, p<0.001). Relatives with Marsh I and Marsh II-III were more often symptomatic (56.3% and 53.8%, respectively) than relatives with normal mucosa (21.1%; p = 0.002). Marsh I relatives had more severe abdominal pain (p = 0.006), severe distension (p = 0.047) and anaemia (p = 0.038) than those with Marsh 0. The prevalence of abnormal bone mineral density was similar in relatives with Marsh I (37%) and Marsh III (44.4%). CONCLUSIONS: The high number of symptomatic patients with lymphocytic enteritis (Marsh I) supports the need for a strategy based on human leucocyte antigen-DQ2 genotyping followed by duodenal biopsy in relatives of patients with coeliac disease and modifies the current concept that villous atrophy is required to prescribe a gluten-free diet. PMID- 16709659 TI - Treatment of oesophageal ulcerations using endoscopic transplantation of tissue engineered autologous oral mucosal epithelial cell sheets in a canine model. AB - BACKGROUND: With the recent development of endoscopic submucosal dissection (ESD), large oesophageal cancers can be removed with a single procedure, with few limits on the resectable range. However, after aggressive ESD, a major complication that arises is postoperative inflammation and stenosis that can considerably affect the patient's quality of life. AIMS: To examine a novel treatment combining ESD and the endoscopic transplantation of tissue-engineered cell sheets created using autologous oral mucosal epithelial cells, in a clinically relevant large animal model. METHODS: Oral mucosal epithelial cells, harvested from beagle dogs, were cultured under normal conditions at 37 degrees C, on temperature-responsive dishes. After ESD (5 cm in length, 180 degrees in range), cell sheets were harvested by a simple reduction in temperature to 20 degrees C, and transplanted by endoscopy. RESULTS: The transplanted cell sheets were able to adhere to and survive on the underlying muscle layers in the ulcer sites, providing an intact, stratified epithelium. Four weeks after surgery, complete wound healing, with no observable stenosis, was seen in the animals receiving autologous cell sheet transplantation. By contrast, noticeable fibrin mesh and host inflammation, consistent with the intermediate stages of wound healing, were observed in the control animals that received only ESD. CONCLUSIONS: These findings in a clinically relevant canine model show the effectiveness of a novel combined endoscopic approach for the potential treatment of oesophageal cancers that can effectively enhance wound healing and possibly prevent postoperative oesophageal stenosis. PMID- 16709660 TI - Clinical presentation, diagnosis and long-term outcome of Wilson's disease: a cohort study. AB - BACKGROUND: Wilson's disease is a rare inborn disease related to copper storage, leading to liver cirrhosis and neuropsychological deterioration. Clinical data on larger cohorts are limited owing to low disease frequency. OBJECTIVE AND METHODS: We performed a retrospective analysis of 163 patients with Wilson's disease, examined at the University of Heidelberg, Heidelberg, Germany, to determine clinical presentation, diagnostic course and long-term outcome. RESULTS: Diagnostic criteria for non-caeruloplasmin-bound serum copper, serum caeruloplasmin, 24-h urinary copper excretion, liver copper content, presence of Kayser-Fleischer rings and histological signs of chronic liver damage were reached in 86.6%, 88.2%, 87.1%, 92.7%, 66.3% and 73% of patients, respectively. By analysis of the coding region of ATP7B (except exons 2, 3 and 21), disease causing mutations were detected in 57% and 29% of patients with Wilson's disease on both chromosomes and on one chromosome, respectively. No mutations were detected in 15% of patients with Wilson's disease. No significant differences were found in clinical parameters or initial presentation between patients grouped according to their mutations. The patients with neurological symptoms were significantly older at the onset of symptoms than patients with hepatitic symptoms (20.2 v 15.5 years of age, p<0.05), and the neurological symptoms were associated with a significantly longer time from onset to diagnosis than hepatic symptoms (44.4 v 14.4 months, p<0.05). After initiating treatment, 76.1% of the patients had a stable or improved course of the disease. Disease progression under treatment was more likely for neuropsychiatric than for hepatic symptoms. Side effects of treatment occurred in 74.4% of patients. CONCLUSIONS: Patients with Wilson's disease having predominantly neuropsychiatric symptoms manifest symptoms later, have a longer time delay from onset of symptoms until definitive diagnosis and have a poorer outcome than patients with hepatic symptoms. PMID- 16709661 TI - Peginterferon alfa-2a (40KD) plus ribavirin in chronic hepatitis C patients who failed previous interferon therapy. AB - BACKGROUND: The management of patients with chronic hepatitis C who have relapsed or failed to respond to interferon based therapies is an important issue facing hepatologists. AIMS: We evaluated the efficacy and safety of peginterferon alfa 2a (40KD) plus ribavirin in this population by conducting a multicentre open label study. PATIENTS: Data from adults with detectable serum hepatitis C virus (HCV) RNA who had not responded or had relapsed after previous conventional interferon or conventional interferon/ribavirin combination therapy were analysed. METHODS: Patients were retreated with peginterferon alfa-2a (40KD) 180 microg/week plus ribavirin 800 mg/day for 24 or 48 weeks at the investigators' discretion. The study was conceived before the optimal dose of ribavirin (1000/1200 mg/day) for patients with genotype 1 was known. The primary endpoint was sustained virological response (SVR), defined as undetectable HCV RNA (<50 IU/ml) after 24 weeks of follow up. The analysis was conducted by intention to treat. RESULTS: A total of 312 patients (212 non-responders, 100 relapsers) were included. Of these, 28 patients were treated for 24 weeks and 284 for 48 weeks. Baseline characteristics between non-responders and relapsers were similar although more non-responders had genotype 1 infection (87% v 69%). Overall SVR rates were 23% (48/212) for non-responders and 41% (41/100) for relapsers. When data were analysed by genotype, SVR rates were 24% (61/253) in genotype 1 and 47% (28/59) in genotype 2/3. CONCLUSIONS: These results in a large patient cohort demonstrate that it is possible to cure a proportion of previous non-responders and relapsers by retreating with peginterferon alfa-2a (40KD) plus ribavirin. PMID- 16709663 TI - The German Bitumen Forum--cooperation in partnership. PMID- 16709662 TI - Soluble galectin-3 is a strong, colonic epithelial-cell-derived, lamina propria fibroblast-stimulating factor. AB - BACKGROUND: Colonic lamina propria fibroblasts (CLPFs) play an important role in the pathogenesis of fibrosis and strictures in Crohn's disease. AIM: To identify colonic epithelial cell (CEC)-derived factors that activate CLPFs. METHODS: Primary human CECs and CLPFs were isolated from control mucosa and interleukin 8 (IL8) of CLPF cultures was quantified by ELISA. Activation of nuclear factor kappaB (NF-kappaB) was shown, and translocation of NF-kappaB was inhibited by a dominant-negative IkappaB-expressing adenovirus. The major CLPF-activating and IL8 inducing protein was purified using fast-performance liquid chromatography (HiPrep 16/60 Sephacryl S-200 High Resolution Column) and sodium dodecyl sulphate gel electrophoresis. RESULTS: A considerable increase in IL8 secretion by CLPFs cultured in CEC-conditioned media compared with that in unconditioned media (155.00 (10.00) pg/microg v 1.434 (0.695) pg/microg) was found. The effect of CEC conditioned media on CLPF IL8 secretion was NF-kappaB dependent. A protein or DNA array confirmed the involvement of NF-kappaB and activator protein-1. Purification of a candidate band isolated with the use of sodium dodecyl sulphate polyacrylamide gel electrophoresis and subsequent sequencing showed soluble galectin-3 to be a strong CLPF-activating factor. Depletion of galectin-3 from conditioned media by immunoprecipitation abolished the CLPF stimulatory effect. CONCLUSIONS: Using a classical biochemical approach, soluble galectin-3 was identified as a strong activator of CLPFs produced by CEC. Galectin-3 induced NF kappaB activation and IL8 secretion in these cells and may be a target for future therapeutic approaches to reduce or avoid stricture formation. PMID- 16709664 TI - WNK1 kinase isoform switch regulates renal potassium excretion. AB - Members of the WNK family of serine/threonine kinases have been implicated as important modulators of salt homeostasis, regulating the balance between renal sodium reabsorption and potassium excretion. Gain-of-expression mutations in the WNK1 gene uncouple Na(+) and K(+) balance and cause a familial disorder of diminished renal potassium excretion, excessive sodium retention, and hypertension (pseudohypoaldosteronism type II or Gordon's syndrome). Alternative splicing of the WNK1 gene produces a kidney-specific short form of WNK1 (KS-WNK1) and a more ubiquitous long form (L-WNK1), but it is not clear how either of these isoforms influence renal potassium excretion. Here we demonstrate that KS-WNK1 and L-WNK1 converge in a pathway to regulate the renal outer-medullary K(+) channel, Kir1.1. Reconstitution studies in Xenopus oocytes reveal that L-WNK1 significantly inhibits Kir1.1 by reducing cell surface localization of the channel. A catalytically inactive L-WNK1 mutant has no inhibitory effect on Kir1.1, indicating that channel inhibition depends on kinase activity. KS-WNK1, lacking an intact kinase domain, does not directly alter Kir1.1. Instead, KS-WNK1 negatively regulates L-WNK1 to release Kir1.1 from inhibition. Acute dietary potassium loading increases the relative abundance of KS-WNK1 to L-WNK1 transcript and protein in the kidney, indicating that physiologic up-regulation of Kir1.1 activity involves a WNK1 isoform switch and KS-WNK1-mediated release from L-WNK1 inhibition. Thus, these observations provide evidence for the physiological regulation of Na(+) and K(+) balance by a kinase isoform switch mechanism. PMID- 16709665 TI - Foxp3+ CD25+ regulatory T cells specific for a neo-self-antigen develop at the double-positive thymic stage. AB - Thymus-derived regulatory T cells (Tregs) expressing CD4, CD25, and the transcription factor Foxp3 play major roles in preventing autoimmunity. The Treg population is enriched in T cells expressing high-avidity self-reactive T cell receptors, and thymic epithelial cells expressing self-antigens (Ag) have been implicated in their induction and/or selection. However, the thymic selection events leading to Treg lineage commitment remain unclear. We followed the thymic development of self-Ag-specific Tregs in double-transgenic mice coexpressing a neo-self-Ag, hemagglutinin (HA) under the control of a neural tissue-specific promoter, and a transgenic class II-restricted T cell antigen receptor specific for HA111-119. Our data show that the promiscuous expression of the HA transgene in thymic epithelial cells is involved in the selective induction and/or expansion of HA-specific Foxp3(+) Treg thymic precursors as early as the double positive stage. PMID- 16709666 TI - Anomalous levels of Cl- transporters in the hippocampal subiculum from temporal lobe epilepsy patients make GABA excitatory. AB - The mRNA levels of NKCC1, an inwardly directed Na(+), K(+)-2Cl(-) cotransporter that facilitates the accumulation of intracellular Cl(-), and of KCC2, an outwardly directed K(+)-Cl(-) cotransporter that extrudes Cl(-), were studied in surgically resected brain specimens from drug-resistant temporal lobe (TL) epilepsy (TLE) patients. Quantitative RT-PCR analyses of the mRNAs extracted from the human TLE-associated brain regions revealed an up-regulation of NKCC1 mRNA and a down-regulation of KCC2 mRNA in the hippocampal subiculum, compared with the hippocampus proper or the TL neocortex, suggesting an abnormal transcription of Cl(-) transporters in the TLE subiculum. In parallel experiments, cell membranes isolated from the same TLE-associated brain regions were injected into Xenopus oocytes that rapidly incorporated human GABA(A) receptors into their surface membrane. The GABA currents elicited in oocytes injected with membranes from the subiculum had a more depolarized reversal potential (E(GABA)) compared with the hippocampus proper or the neocortex. The NKCC1 blocker bumetanide or a temperature decrease of 10 degrees C shifted the GABA-current E(GABA) more negative in oocytes injected with membranes from TLE hippocampal subiculum, matching the E(GABA) of TL neocortex-injected oocytes. We conclude that the anomalous expression of both Cl(-) transporters, NKCC1 and KCC2 [corrected] in TLE hippocampal subiculum probably causes altered Cl(-) transport in the "epileptic" neurons, as revealed in the microtransplanted Xenopus oocytes, and renders GABA aberrantly "exciting," a feature that may contribute to the precipitation of epileptic seizures. PMID- 16709667 TI - Efficient gene delivery to pancreatic islets with ultrasonic microbubble destruction technology. AB - This study describes a method of gene delivery to pancreatic islets of adult, living animals by ultrasound targeted microbubble destruction (UTMD). The technique involves incorporation of plasmids into the phospholipid shell of gas filled microbubbles, which are then infused into rats and destroyed within the pancreatic microcirculation with ultrasound. Specific delivery of genes to islet beta cells by UTMD was achieved by using a plasmid containing a rat insulin 1 promoter (RIP), and reporter gene expression was regulated appropriately by glucose in animals that received a RIP-luciferase plasmid. To demonstrate biological efficacy, we used UTMD to deliver RIP-human insulin and RIP-hexokinase I plasmids to islets of adult rats. Delivery of the former plasmid resulted in clear increases in circulating human C-peptide and decreased blood glucose levels, whereas delivery of the latter plasmid resulted in a clear increase in hexokinase I protein expression in islets, increased insulin levels in blood, and decreased circulating glucose levels. We conclude that UTMD allows relatively noninvasive delivery of genes to pancreatic islets with an efficiency sufficient to modulate beta cell function in adult animals. PMID- 16709668 TI - The AAA ATPase p97 links peptide N-glycanase to the endoplasmic reticulum associated E3 ligase autocrine motility factor receptor. AB - Mouse peptide N-glycanase (mPNGase) cleaves the N-glycan chain from misfolded glycoproteins and glycopeptides. Previously, several proteins were found to directly interact with mPNGase; among them, both mHR23B and mS4 were found to link mPNGase to the proteasome. In this study, we found that the cytoplasmic protein mp97 participates in the formation of a ternary complex containing mouse autocrine motility factor receptor (mAMFR), mp97, and mPNGase. This assemblage recruits the cytosolic mPNGase close to the endoplasmic reticulum (ER) membrane, where the retrotranslocation of misfolded glycoproteins is thought to occur. In addition to the ER membrane-associated E3 ligase mAMFR, a cytosolic protein mY33K, containing both UBA and UBX domains, was found to also directly interact with mp97. Thus, a complex containing five proteins, mAMFR, mY33K, mp97, mPNGase, and mHR23B, is formed in close proximity to the ER membrane and serves to couple the activities of retrotranslocation, ubiquitination, and deglycosylation and, thereby, route misfolded glycoproteins to the proteasome. PMID- 16709669 TI - Processes of copy-number change in human DNA: the dynamics of {alpha}-globin gene deletion. AB - Ectopic recombination between locally repeated DNA sequences is of fundamental importance in the evolution of gene families, generating copy-number variation in human DNA and often leading to pathological rearrangements. Despite its importance, little is known about the dynamics and processes of these unequal crossovers and the degree to which meiotic recombination plays a role in instability. We address this issue by using as a highly informative system the duplicated alpha-globin genes in which ectopic recombination can lead to gene deletions, often very prevalent in populations affected by malaria, as well as reduplications. Here we show that spontaneous deletions can be accessed directly in genomic DNA by using single-DNA-molecule methods. These deletions proved to be remarkably common in both blood and sperm. Somatic deletions arise by a strictly intrachromosomal pathway of homologous exchange that also operates in the germ line and can generate mutational mosaicism, whereas sperm deletions frequently involve recombinational interactions between homologous chromosomes that most likely occur at meiosis. Ectopic recombination frequencies show surprisingly little requirement for long, identical homology blocks shared by paralogous sequences, and exchanges can occur even between short regions of sequence identity. Finally, direct knowledge of germ-line deletion rates can give insights into the fitness of individuals with these alpha-globin gene deletions, providing a new approach to investigating historical levels of selection operating in human populations. PMID- 16709670 TI - Structural and functional recovery from early monocular deprivation in adult rats. AB - Visual deficits caused by abnormal visual experience during development are hard to recover in adult animals. Removal of chondroitin sulfate proteoglycans from the mature extracellular matrix with chondroitinase ABC promotes plasticity in the adult visual cortex. We tested whether chondroitinase ABC treatment of adult rats facilitates anatomical, functional, and behavioral recovery from the effects of a period of monocular deprivation initiated during the critical period for monocular deprivation. We found that chondroitinase ABC treatment coupled with reverse lid-suturing causes a complete recovery of ocular dominance, visual acuity, and dendritic spine density in adult rats. Thus, manipulations of the extracellular matrix can be used to promote functional recovery in the adult cortex. PMID- 16709671 TI - Conformation of the synaptobrevin transmembrane domain. AB - The synaptic vesicle protein synaptobrevin (also called VAMP, vesicle-associated membrane protein) forms part of the SNARE (soluble N-ethylmaleimide sensitive factor attachment protein receptor) complex, which is essential for vesicle fusion. Additionally, the synaptobrevin transmembrane domain can promote lipid mixing independently of complex formation. Here, the conformation of the transmembrane domain was studied by using circular dichroism and attenuated total reflection Fourier-transform infrared spectroscopy. The synaptobrevin transmembrane domain has an alpha-helical structure that breaks in the juxtamembrane region, leaving the cytoplasmic domain unstructured. In phospholipid bilayers, infrared dichroism data indicate that the transmembrane domain adopts a 36 degrees angle with respect to the membrane normal, similar to that reported for viral fusion peptides. A conserved aromatic/basic motif in the juxtamembrane region may be causing this relatively high insertion angle. PMID- 16709673 TI - Dynamic fragility of oceanic coral reef ecosystems. AB - As one of the most diverse and productive ecosystems known, and one of the first ecosystems to exhibit major climate-warming impacts (coral bleaching), coral reefs have drawn much scientific attention to what may prove to be their Achilles heel, the thermal sensitivity of reef-building corals. Here we show that climate change-driven loss of live coral, and ultimately structural complexity, in the Seychelles results in local extinctions, substantial reductions in species richness, reduced taxonomic distinctness, and a loss of species within key functional groups of reef fish. The importance of deteriorating physical structure to these patterns demonstrates the longer-term impacts of bleaching on reefs and raises questions over the potential for recovery. We suggest that isolated reef systems may be more susceptible to climate change, despite escaping many of the stressors impacting continental reefs. PMID- 16709672 TI - Effects of surface tethering on protein folding mechanisms. AB - The folding mechanisms of proteins are increasingly being probed through single molecule experiments in which the protein is immobilized on a surface. Nevertheless, a clear understanding of how the surface might affect folding, and whether or not it changes folding from its bulk behavior, is lacking. In this work, we use molecular dynamics simulations of a model beta-barrel protein tethered to a surface to systematically investigate how the surface impacts folding. In the bulk, this protein folds in a three-state manner through a compact intermediate state, and its transition state (TS) has a well formed hydrophobic core. Upon tethering, we find that folding rates and stability are impacted differently by the surface, with dependencies on both the length and location of the tether. Significant changes in folding times are observed for tether points that do not alter the folding temperature. Tethering also locally enhances the formation of structure for residues proximal to the tether point. We find that neither the folding mechanism nor the TS of this protein are altered if the tether is in a fully structured or completely unstructured region of the TS. By contrast, tethering in a partially structured region of the TS leads to dramatic changes. For one such tether point, the intermediate present in bulk folding is eliminated, leading to a two-state folding process with a heterogeneous, highly unstructured TS ensemble. These results have implications for both the design of single-molecule experiments and biotechnological applications of tethered proteins. PMID- 16709674 TI - Postsynaptic inositol 1,4,5-trisphosphate signaling maintains presynaptic function of parallel fiber-Purkinje cell synapses via BDNF. AB - The maintenance of synaptic functions is essential for neuronal information processing, but cellular mechanisms that maintain synapses in the adult brain are not well understood. Here, we report an activity-dependent maintenance mechanism of parallel fiber (PF)-Purkinje cell (PC) synapses in the cerebellum. When postsynaptic metabotropic glutamate receptor (mGluR) or inositol 1,4,5 trisphosphate (IP(3)) signaling was chronically inhibited in vivo, PF-PC synaptic strength decreased because of a decreased transmitter release probability. The same effects were observed when PF activity was inhibited in vivo by the suppression of NMDA receptor-mediated inputs to granule cells. PF-PC synaptic strength similarly decreased after the in vivo application of an antibody against brain-derived neurotrophic factor (BDNF). Furthermore, the weakening of synaptic connection caused by the blockade of mGluR-IP(3) signaling was reversed by the in vivo application of BDNF. These results indicate that a signaling cascade comprising PF activity, postsynaptic mGluR-IP(3) signaling and subsequent BDNF signaling maintains presynaptic functions in the mature cerebellum. PMID- 16709675 TI - Assessing implicit models for nonpolar mean solvation forces: the importance of dispersion and volume terms. AB - Continuum solvation models provide appealing alternatives to explicit solvent methods because of their ability to reproduce solvation effects while alleviating the need for expensive sampling. Our previous work has demonstrated that Poisson Boltzmann methods are capable of faithfully reproducing polar explicit solvent forces for dilute protein systems; however, the popular solvent-accessible surface area model was shown to be incapable of accurately describing nonpolar solvation forces at atomic-length scales. Therefore, alternate continuum methods are needed to reproduce nonpolar interactions at the atomic scale. In the present work, we address this issue by supplementing the solvent-accessible surface area model with additional volume and dispersion integral terms suggested by scaled particle models and Weeks-Chandler-Andersen theory, respectively. This more complete nonpolar implicit solvent model shows very good agreement with explicit solvent results and suggests that, although often overlooked, the inclusion of appropriate dispersion and volume terms are essential for an accurate implicit solvent description of atomic-scale nonpolar forces. PMID- 16709676 TI - The nonredundant roles of two 4'-phosphopantetheinyl transferases in vital processes of Mycobacteria. AB - Mycobacterium tuberculosis contains >20 enzymes that require activation by transfer of the 4'-phosphopantetheine moiety of CoA onto a conserved serine residue, a posttranslational modification catalyzed by 4'-phosphopantetheinyl transferases (PPTases). The modified proteins are involved in key metabolic processes such as cell envelope biogenesis and the production of virulence factors. We show that two PPTases conserved in all Mycobacterium spp. and in related genera activate two different subsets of proteins and are not functionally redundant. One enzyme, AcpS, activates the two fatty acid synthase systems of mycobacteria, whereas the other PPTase, PptT, acts on type-I polyketide synthases and nonribosomal peptide synthases, both of which are involved in the biosynthesis of virulence factors. We demonstrate that both PPTases are essential for Mycobacterium smegmatis viability and that PptT is required for the survival of Mycobacterium bovis bacillus Calmette-Guerin. These enzymes are thus central to the biology of mycobacteria and for mycobacterial pathogenesis and represent promising targets for new antituberculosis drugs. PMID- 16709678 TI - Arundic Acid ameliorates cerebral amyloidosis and gliosis in Alzheimer transgenic mice. AB - Like microglia, reactive astrocytes produce a myriad of neurotoxic substances in various brain pathologies, such as Alzheimer's disease (AD), trauma, and cerebral ischemia. Among the numerous products of reactive astrocytes, attention has recently been directed toward the possible detrimental role of S100B, because the protein has been shown to be highly expressed along with the progression of brain damage and to exert neurotoxic effects at high concentrations. The present study aimed to examine the possible role of astrocyte-derived S100B in the progression of cerebral amyloidosis and gliosis in transgenic mice overproducing mutant amyloid precursor protein (Tg APP(sw) mice, line 2576). For this purpose, arundic acid (Ono Pharmaceutical Co., Ltd., Mishima, Osaka, Japan), which is known to negatively regulate astrocyte synthesis of S100B, was orally administered to Tg APP(sw) mice for 6 months from 12 months of age, and the effects of the agent on the above parameters were examined. Here, we report that beta-amyloid deposits along with amyloid-beta peptide/S100B levels, as well as beta-amyloid plaque associated reactive gliosis (astrocytosis and microgliosis), were significantly ameliorated in arundic acid-treated Tg APP(sw) mice relative to vehicle-treated Tg APP(sw) mice at 19 months of age. Based on the above results, arundic acid is considered to deserve further exploration as a promising therapeutic agent for AD. PMID- 16709677 TI - A unique strategy for mRNA cap methylation used by vesicular stomatitis virus. AB - Nonsegmented negative-sense (nsNS) RNA viruses typically replicate within the host cell cytoplasm and do not have access to the host mRNA capping machinery. These viruses have evolved a unique mechanism for mRNA cap formation in that the guanylyltransferase transfers GDP rather than GMP onto the 5' end of the RNA. Working with vesicular stomatitis virus (VSV), a prototype nsNS RNA virus, we now provide genetic and biochemical evidence that its mRNA cap methylase activities are also unique. Using recombinant VSV, we determined the function in mRNA cap methylation of a predicted binding site in the polymerase for the methyl donor, S adenosyl-l-methionine. We found that amino acid substitutions to this site disrupted methylation at the guanine-N-7 (G-N-7) position or at both the G-N-7 and ribose-2'-O (2'-O) positions of the mRNA cap. These studies provide genetic evidence that the two methylase activities share an S-adenosyl-l-methionine binding site and show that, in contrast to other cap methylation reactions, methylation of the G-N-7 position is not required for 2'-O methylation. These findings suggest that VSV evolved an unusual strategy of mRNA cap methylation that may be shared by other nsNS RNA viruses. PMID- 16709679 TI - Diadenosine polyphosphate analog controls postsynaptic excitation in CA3-CA1 synapses via a nitric oxide-dependent mechanism. AB - Previously, we have described the modulatory effect of diadenosine polyphosphates Ap4A and Ap5A on synaptic transmission in the rat hippocampal slices mediated by presynaptic receptors (Klishin et al., 1994). In contrast, we now describe how nonhydrolyzable Ap4A analog diadenosine-5',5'''-P1,P4-[beta,beta' methylene]tetraphosphate (AppCH2ppA) at low micromolar concentrations exerts strong nondesensitizing inhibition of orthodromically evoked field potentials (OFPs) without affecting the amplitude of excitatory postsynaptic currents and antidromically evoked field potentials, as recorded in hippocampal CA1 zone. The effects of AppCH2ppA on OFPs are eliminated by a P2 receptor antagonist pyridoxal phosphate-6-azophenyl-2',4'-disulfonic acid (PPADS) but not mimicked by purinoceptor agonists alpha,beta-methylene-ATP and adenosine 5'-O-(3-thio) triphosphate, indicating that a P2-like receptor is involved but not one belonging to the conventional P2X/P2Y receptor classes. Diadenosine polyphosphate receptor (P4) antagonist Ip4I (diinosine tetraphosphate) was unable to modulate AppCH2ppA effects. Thus, the PPADS-sensitive P2-like receptor for AppCH2ppA seems to control selectively dendritic excitation of the CA1 neurons. The specific nitric oxide (NO)-scavenger 2-phenyl-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3 oxide is shown to significantly attenuate AppCH2ppA-mediated inhibitory effects, indicating that NO is involved in the cascade of events initiated by AppCH2ppA. Further downstream mediation by adenosine A1 receptors is also demonstrated. Hence, AppCH2ppA-mediated effects involve PPADS-sensitive P2-like receptor activation leading to the production of NO that stimulates intracellular synthesis of adenosine, causing in turn postsynaptic A1 receptor activation and subsequent postsynaptic CA1 dendritic inhibition. Such spatially selective postsynaptic dendritic inhibition may influence dendritic electrogenesis in pyramidal neurons and consequently mediate control of neuronal network activity. PMID- 16709680 TI - Carpal box and open cup radiography. PMID- 16709681 TI - Back to the basics. PMID- 16709683 TI - Advanced CT technologist needed. PMID- 16709682 TI - X-ray technique in CR. PMID- 16709685 TI - Admission requirements for radiography programs. AB - CONTEXT: Radiography program applicants far outnumber available class capacity in many programs. Educational accountability requires that programs justify their admission practices, work to retain all qualified students and evaluate their attrition rates. Thus, radiography programs are interested in identifying the best candidates for admission. PURPOSE: The purpose of this small survey study was to document admission requirements of entry-level programs in radiography accredited by the Joint Review Committee on Education in Radiologic Technology (JRCERT) in Oklahoma and Texas. METHODS: A 14-question survey was sent to 45 JRCERT-accredited entry-level radiography programs in Oklahoma and Texas. The survey included questions concerning program demographics, admission criteria and admission practices. RESULTS: Cumulative grade point average, interviews, successful completion of math and science courses, and performance on standardized tests were the most commonly used admission criteria, although no criteria were used by all of the programs surveyed. CONCLUSION: The current study is the first step toward identifying the most effective admission criteria for radiography programs. Future research should examine the correlation of specific admission criteria with program success (ie, program retention, program completion and professional certification). PMID- 16709684 TI - The magic-angle effect of the supraspinatus tendon. AB - CONTEXT: Rotator cuff injuries are common and result in either a tear or tendinopathy within the distal portion of the supraspinatus tendon. Magnetic resonance (MR) imaging shows the injury as a high signal region within the distal tendon. Similarly, a magic-angle effect also appears within healthy patients in this same region on T1- or proton density (PD)-weighted MR sequences. OBJECTIVE: This study was conducted to evaluate the prevalence of the magic-angle effect found with MR imaging within the supraspinatus tendon. METHODS: In this study, 300 consecutive patients were selected from those symptomatic patients referred for MR evaluation of the shoulder using a 1.5T unit and both PD and T2-weighted sequences. To eliminate patients with a real injury, the written reports were reviewed; those with positive findings for injury to the supraspinatus tendon were eliminated from the sample group, leaving 164 patients. Two radiographers independently examined the oblique coronal images nearest the center of the supraspinatus tendon and compared the signal on PD and T2-weighted images. Images found to have a higher signal with the PD sequence were classified as having the magic-angle effect since a real injury would be more likely to have a strong signal on T2-weighted images. RESULTS: Of the 164 patients evaluated, both reviewers found the same 8 patients to have the magic-angle effect. SUMMARY: Based on these results, our study suggests the artifact will appear in 5% of healthy patients and may lead to false-positive results on oblique coronal PD weighted images through the supraspinatus tendon. In comparison with previous studies that have shown a much greater incidence rate, our findings also suggest that external rotation of the arm greatly reduces the incidence of the magic angle effect. PMID- 16709686 TI - Diagnostic reference levels in radiology. PMID- 16709687 TI - Pelvic organ prolapse. AB - Pelvic organ prolapse, a condition in which the ligaments and muscles that suspend the vagina within the pelvic cavity weaken or break, is a frequent cause of urinary and fecal incontinence. Stigma, embarrassment and the belief that pelvic organ prolapse is a natural part of aging prevents many women from seeking treatment. Medical imaging modalities such as defecography, dynamic magnetic resonance imaging and ultrasound help health care providers make effective treatment decisions. PMID- 16709689 TI - Uncommon findings on ultrasound. Rib fractures. PMID- 16709691 TI - Patient page. Upper GI series. PMID- 16709692 TI - Mitral regurgitation and left ventricular dyssynchrony: implications for treatment. AB - Cardiac resynchronisation therapy may provide an alternative treatment for mitral regurgitation in patients unsuitable for surgery. PMID- 16709693 TI - Hospital volume of throughput and periprocedural and medium-term adverse events after percutaneous coronary intervention: retrospective cohort study of all 17,417 procedures undertaken in Scotland, 1997-2003. AB - OBJECTIVE: To determine whether percutaneous coronary intervention (PCI) hospital volume of throughput is associated with periprocedural and medium-term events, and whether any associations are independent of differences in case mix. DESIGN: Retrospective cohort study of all PCIs undertaken in Scottish National Health Service hospitals over a six-year period. METHODS: All PCIs in Scotland during 1997-2003 were examined. Linkage to administrative databases identified events over two years' follow up. The risk of events by hospital volume at 30 days and two years was compared by using logistic regression and Cox proportional hazards models. RESULTS: Of the 17,417 PCIs, 4900 (28%) were in low-volume hospitals and 3242 (19%) in high-volume hospitals. After adjustment for case mix, there were no significant differences in risk of death or myocardial infarction. Patients treated in high-volume hospitals were less likely to require emergency surgery (adjusted odds ratio 0.18, 95% confidence interval (CI) 0.07 to 0.54, p = 0.002). Over two years, patients in high-volume hospitals were less likely to undergo surgery (adjusted hazard ratio 0.52, 95% CI 0.35 to 0.75, p = 0.001), but this was offset by an increased likelihood of further PCI. There was no net difference in coronary revascularisation or in overall events. CONCLUSION: Death and myocardial infarction were infrequent complications of PCI and did not differ significantly by volume. Emergency surgery was less common in high-volume hospitals. Over two years, patients treated in high-volume centres were as likely to undergo some form of revascularisation but less likely to undergo surgery. PMID- 16709694 TI - Distribution of SPARC during neovascularisation of degenerative aortic stenosis. AB - OBJECTIVE: To examine the hypothesis that degenerative aortic stenosis (AS) is associated with the development of blood vessels and the expression of the secreted protein, acidic and rich in cysteine/osteonectin (SPARC), a matricellular protein that is involved in ossification, the modulation of angiogenesis and the production of metalloproteinases. METHODS: 30 surgically excised AS valves and 20 normal aortic valves were studied. RESULTS: Blood vessels were detected in the aortic valves from patients with degenerative AS, whereas normal valves were avascular structures. Blood vessels in AS valves expressed endothelial nitric oxide synthase, CD34 and von Willebrand factor (vWF). Blood vessels were located in three distinct regions: near calcified nodules, under the leaflet border and in rich cellular areas forming cell islands. Blood vessels were predominantly present in early and intermediate grades of calcification. Cell islands were densely populated by CD45-positive cells where endothelial cells (CD34+, vWF+) forming cord-like structures were present. Immunoblotting detected SPARC only in AS valves and immunohistological analysis located SPARC in mature blood vessels. The proportion of blood vessels positive for SPARC was higher in valves with a lower grade of calcification. In cell islands, SPARC was distributed to mature blood vessels and to macrophages, where it co-located with matrix metalloproteinase-9, whereas no expression was detected in endothelial cells forming cord-like structures. CONCLUSION: The localisation of SPARC to mature blood vessels and its predominant expression in AS valves with a lower calcification grade suggest that the spatial and temporal distribution of this matricellular protein is tightly controlled to participate in the neovascularisation of AS valves. PMID- 16709695 TI - Assessment of cardiac risk before non-cardiac surgery: brain natriuretic peptide in 1590 patients. AB - OBJECTIVE: To evaluate the predictive value of brain natriuretic peptide (BNP) for assessment of cardiac risk before non-cardiac surgery. METHODS: Consecutively treated patients (947 men, 643 women) whose BNP was measured before non-cardiac surgery were studied. Clinical and ECG variables were evaluated to identify predictors of postoperative cardiac events. RESULTS: Events occurred in 6% of patients: 21 cardiac deaths, 20 non-fatal myocardial infarctions, 41 episodes of pulmonary oedema and 14 patients with ventricular tachycardia. All of these patients had raised plasma BNP concentrations (best cut-off point 189 pg/ml). The only independent predictor of postoperative events was BNP (odds ratio 34.52, 95% confidence interval (CI) 17.08 to 68.62, p < 0.0001). Clinical variables of Goldman's multifactorial index identified 18% of patients in class I, 40% in class II, 24% in class III and 18% in class IV preoperatively; postoperative event rates were 2%, 3%, 7% and 14%, respectively. BNP identified 60% of patients as having zero risk (BNP 0-100 pg/ml), 22% low risk (101-200 pg/ml), 14% intermediate risk (201-300 pg/ml) and 4% high risk (> 300 pg/ml); postoperative event rates were 0%, 5%, 12% and 81%, respectively. CONCLUSIONS: In this population of patients evaluated before non-cardiac surgery, BNP is an independent predictor of postoperative cardiac events. BNP > 189 pg/ml identified patients at highest risk. PMID- 16709696 TI - Prolonged mechanical systole and increased arterial wave reflections in diastolic dysfunction. AB - OBJECTIVE: To evaluate whether left ventricular ejection time indexed for heart rate (left ventricular ejection time index (LVETI)) and arterial wave reflections (augmented pressure (AP)) are increased in patients with diastolic dysfunction (DD). DESIGN: Prospective observational study. SETTING: University teaching hospital providing primary and tertiary care. SUBJECTS: 235 consecutive patients undergoing left heart catheterisation were categorised as having definite DD, possible DD or no DD (controls) on the basis of their left ventricular end diastolic pressures and N-terminal brain natriuretic peptide concentrations. MAIN OUTCOME MEASURES: LVETI and AP were prospectively assessed non-invasively by radial applanation tonometry. In addition, all patients underwent comprehensive echocardiography, including tissue Doppler imaging of mitral annulus velocity in early diastole (E'). RESULTS: LVETI was longer in patients with definite DD than in patients with possible DD and in controls (433.6 (SD 17.2), 425.9 (17.9) and 414.3 (13.6) ms, respectively, p < 0.000001). Arterial wave reflections were higher in definite DD than in possible DD and control groups (AP was 19.4 (SD 8.9), 15.2 (8.0) and 10.7 (6.8) mm Hg, respectively, p < 0.000001). In receiver operating characteristic curve analysis, LVETI detected DD as well as echocardiography (E:E'). Area under the curve for LVETI to differentiate patients with definite DD from normal controls was 0.81 (95% CI 0.72 to 0.89, p < 0.0001). In multivariable logistic regression analysis, LVETI added significant independent power to clinical and echocardiographic variables for prediction of DD. CONCLUSIONS: Mechanical systole is prolonged and arterial wave reflections are increased in most patients with DD. Rapid non-invasive assessment of these parameters may aid in confirming or excluding DD. PMID- 16709698 TI - Haemodynamic effects of changes in atrioventricular and interventricular delay in cardiac resynchronisation therapy show a consistent pattern: analysis of shape, magnitude and relative importance of atrioventricular and interventricular delay. AB - OBJECTIVE: To assess the haemodynamic effect of simultaneously adjusting atrioventricular (AV) and interventricular (VV) delays. METHOD: 35 different combinations of AV and VV delay were tested by using digital photoplethysmography (Finometer) with repeated alternations to measure relative change in systolic blood pressure (SBP(rel)) in 15 patients with cardiac resynchronisation devices for heart failure. RESULTS: Changing AV delay had a larger effect than changing VV delay (range of SBP(rel) 21 v 4.2 mm Hg, p < 0.001). Each had a curvilinear effect. The curve of response to AV delay fitted extremely closely to a parabola (average R2 = 0.99, average residual variance 0.8 mm Hg2). The response to VV delay was significantly less curved (quadratic coefficient 67 v 1194 mm Hg/s2, p = 0.003) and therefore, although the residual variance was equally small (0.8 mm Hg2), the R2 value was 0.7. Reproducibility at two months was good, with the SD of the difference between two measurements of SBP(rel) being 2.5 mm Hg for AV delay (2% of mean systolic blood pressure) and 1.5 mm Hg for VV delay (1% of mean systolic blood pressure). CONCLUSIONS: Changing AV and VV delays results in a curvilinear acute blood pressure response. This shape fits very closely to a parabola, which may be valuable information in developing a streamlined clinical protocol. VV delay adjustment provides an additional, albeit smaller, haemodynamic benefit to AV optimisation. PMID- 16709697 TI - Short-term statin treatment improves endothelial function and neurohormonal imbalance in normocholesterolaemic patients with non-ischaemic heart failure. AB - OBJECTIVES: To investigate the effect of short-term statin treatment on impaired endothelium-dependent vasodilatation and haemodynamic abnormalities typically occurring in chronic heart failure (CHF). METHODS: In a double-blind, crossover study endothelium-dependent vasodilatation was measured in conduit and resistance vessels of 23 patients with non-ischaemic CHF after 6 weeks of placebo and 40 mg atorvastatin. The haemodynamic impact was assessed by cardioendocrine hormones, echocardiography and clinical indicators of CHF. RESULTS: Cholesterol concentrations were population average (low density lipoprotein 3.56 (SEM 0.16) mmol/l, triglycerides 1.70 (0.20) mmol/l and high density lipoprotein 1.17 (0.07) mmol/l). In resistance vessels, the area under the curve ratio during acetylcholine infusion increased from 9.2 (1.9) with placebo to 12.2 (2.1) with statin (p < 0.01). This improvement was reversed during co-infusion with the nitric oxide antagonist N(G)-monomethyl-L-arginine. In conduit arteries, flow mediated dilatation increased from 5.64 (SEM 0.88)% with placebo to 6.83 (0.97)% with statin (p < 0.05). Endothelium-independent vasodilatation did not change (p = 0.68 for conduit and p = 0.45 for resistance vessels). Endothelin 1 and atrial natriuretic peptide (ANP) decreased from 1.57 (0.08) and 51.3 (1.0) with placebo to 1.42 (0.09) pg/ml (p < 0.05) and 42.1 (7.5) pmol/l (p < 0.05), respectively, with statin. CONCLUSIONS: In patients with non-ischaemic CHF and population average cholesterol concentrations, short-term statin treatment improves endothelial function in conduit and resistance vessels and lowers plasma endothelin 1 and ANP concentrations. PMID- 16709699 TI - Predictors of patients' preferences for treatments to prevent heart disease. AB - OBJECTIVES: To determine the relationship between expressed preferences for drug treatment to prevent coronary disease and several participant and general practitioner characteristics among patients attending coronary risk screening. DESIGN: Face-to-face interviews with patients. At the first interview, a researcher asked participants to imagine six scenarios representing different levels of pretreatment five-year coronary risk. In each case they were asked whether they would choose treatment that would reduce their coronary risk by 30% of pretreatment risk. At the second interview participants were told their coronary risk and asked whether they would choose treatment. Sociodemographic variables were collected to investigate their relationship to patients' treatment preferences. PARTICIPANTS: Patients identified as likely to be at high coronary risk were invited to attend for risk screening and to participate in the study. SETTING: 13 practices in the West Midlands. RESULTS: Participants' preferences varied widely: at the first interview 112 (55.2%) of 203 participants preferred treatment at 3% five-year coronary risk but 31 (15.3%) preferred no treatment even at 30% five-year coronary risk. Age, sex, education and drug treatment history did not affect preferences, but lower social class was associated with preferring treatment at lower risk. Preferences expressed at the second interview were generally consistent with preferences at the first interview (kappa = 0.510, 95% CI 0.380 to 0.639). CONCLUSIONS: Patients attending for coronary risk screening express stable preferences for drug treatment to prevent coronary heart disease. Their preferences vary widely and may be associated with social class. PMID- 16709700 TI - Promotion of proinflammatory interactions between platelets and monocytes by unfractionated heparin. AB - OBJECTIVES: To determine the in vitro effects of unfractionated heparin, fractionated heparin and direct thrombin inhibition on platelet-monocyte aggregation, and to establish the in vivo effects of unfractionated heparin and direct thrombin inhibition on platelet-monocyte aggregates in patients scheduled for percutaneous coronary intervention (PCI). DESIGN: Platelet-monocyte aggregates were assessed in whole blood from 18 healthy volunteers after the addition of unfractionated heparin (1 U/ml), enoxaparin (0.8 U/ml) or lepirudin (5.6 microg/ml), and in 28 patients scheduled for elective PCI before and after administration of 100 U/kg of unfractionated heparin or 0.75 mg/kg bivalirudin. The influence of P-selectin-mediated platelet-monocyte aggregation was assessed with specific blocking antibodies. RESULTS: Addition of unfractionated heparin in vitro was associated with a higher level of platelet-monocyte aggregates than in controls (20.1 (1.9)% v 16.2 (1.6)%, respectively, p < 0.001). However, platelet monocyte aggregation was not affected by enoxaparin or lepirudin (16.9 (2.0)% and 17.0 (2.2)%, respectively, NS). Intravenous unfractionated heparin in vivo also resulted in an increase in platelet-monocyte aggregates (absolute Delta 7.1 (2.7)%, p < 0.01), whereas intravenous bivalirudin had no effect (absolute Delta 1.5 (2.4)%, NS). The addition of P-selectin blockade abolished any increase in platelet-monocyte aggregates associated with heparin. CONCLUSIONS: In vitro and in vivo unfractionated heparin is associated with increased platelet-monocyte aggregation through a P-selectin-dependent mechanism. These findings provide a potential explanation for the superior cardiovascular outcomes associated with fractionated heparins and direct thrombin inhibitors. PMID- 16709701 TI - Physical capacity in relation to low back, neck, or shoulder pain in a working population. AB - AIMS: To investigate the longitudinal relation between physical capacity (isokinetic lifting strength, static endurance of the back, neck, and shoulder muscles, and mobility of the spine) and low back, neck, and shoulder pain. METHODS: In this prospective cohort study, 1789 Dutch workers participated. At baseline, isokinetic lifting strength, static endurance of the back, neck, and shoulder muscles, and mobility of the spine were measured in the pain free workers, as well as potential confounders, including physical workload. Low back, neck, and shoulder pain were self-reported annually at baseline and three times during follow up. RESULTS: After adjustment for confounders, Poisson generalised estimation equations showed an increased risk of low back pain among workers in the lowest sex specific tertile of performance in the static back endurance tests compared to workers in the reference category (RR = 1.42; 95% CI 1.19 to 1.71), but this was not found for isokinetic trunk lifting strength or mobility of the spine. An increased risk of neck pain was shown for workers with low performance in tests of isokinetic neck/shoulder lifting strength (RR = 1.31; 95% CI 1.03 to 1.67) and static neck endurance (RR = 1.22; 95% CI 1.00 to 1.49). Among workers in the lowest tertiles of isokinetic neck/shoulder lifting strength or endurance of the shoulder muscles, no increased risk of shoulder pain was found. CONCLUSIONS: The findings of this study suggest that low back or neck endurance were independent predictors of low back or neck pain, respectively, and that low lifting neck/shoulder strength was an independent predictor of neck pain. No association was found between lifting trunk strength, or mobility of the spine and the risk of low back pain, nor between lifting neck/shoulder strength or endurance of the shoulder muscles and the risk of shoulder pain. PMID- 16709702 TI - The triad of shift work, occupational noise, and physical workload and risk of coronary heart disease. AB - BACKGROUND: Shift work, noise, and physical workload are very common occupational exposures and they tend to cluster in the same groups of workers. OBJECTIVES: To study the short and long term effects of these exposures on risk of coronary heart disease (CHD) and to estimate the joint effects of these factors. METHODS: The study population in this prospective 13 year follow up study of 1804 middle aged industrially employed men was collected at the first screening for the Helsinki Heart Study. The CHD end points (ICD-9 codes 410-414 and ICD-10 codes I20-I25) were obtained from official Finnish registers. The Finnish job-exposure matrix FINJEM provided information on occupational exposures. Relative risks (RR) of CHD for the exposures were estimated using Cox's proportional hazard models adjusting for classical risk factors of CHD. RESULTS: The RR in the five year follow up for continuous noise combined with impulse noise was 1.28; for shift work it was 1.59, and for physical workload 1.18, while in the 13 year follow up the RRs were 1.58, 1.34, and 1.31, respectively. When adjusted for white collar/blue-collar status the RRs decreased markedly. The RR in the 13 year follow up for those exposed to two risk factors was close to 1.7 and for those exposed to all three, 1.87. CONCLUSION: Shift work and continuous noise entailed an excess risk for CHD in the shortest follow up with only a few retired workers but a decreasing risk during the longer follow up. For physical workload and impulse noise the trend was opposite: the CHD risk was increasing with increasing follow up time despite increasing numbers of retired workers. PMID- 16709703 TI - The risk for multiple sclerosis in female nurse anaesthetists: a register based study. AB - BACKGROUND: Previous studies have suggested that exposure to organic solvents, including volatile anaesthetic agents, may be a risk factor for multiple sclerosis (MS), possibly in combination with genetic and other environmental factors. AIMS: To further investigate the role of volatile anaesthetic agents having similar acute toxic effects to other organic solvents. METHODS: Female nurse anaesthetists, other female nurses, and female teachers from middle and upper compulsory school levels were identified and retrieved from the 1985 census, Statistics Sweden. By means of the unique personal identity number in Sweden, these individuals were linked with the disability pension registers at The National Social Insurance Board and also with data on hospital care 1985-2000 at The National Board of Health and Welfare. RESULTS: The cumulative incidence rate ratio of MS was found to be increased in female nurse anaesthetists in relation to other nurses (statistically not significant) and teachers (statistically significant), respectively. CONCLUSIONS: These findings give some support to previous findings of an increased risk for MS in nurse anaesthetists. This is interesting in the context of previous observations of organic solvents in general as a potential risk factor in MS. PMID- 16709704 TI - Global quality control perspective for the physical and technical aspects of screen-film mammography--image quality and radiation dose. AB - The systematic monitoring of image quality and radiation dose is an ultimate solution to ensuring the continuously high quality of mammography examination. At present several protocols exist around the world, and different test objects are used for quality control (QC) of the physical and technical aspects of screen film mammography. This situation may lead to differences in radiation image quality and dose reported. This article reviews the global QC perspective for the physical and technical aspects of screen-film mammography with regard to image quality and radiation dose. It points out issues that must be resolved in terms of radiation dose and that also affect the comparison. PMID- 16709705 TI - Space radiation dosimetry by combination of integrating dosemeters. AB - Generally, individual monitoring is performed with a small personal dosemeter which responds to radiation by giving a necessary operational quantity such as personal dose equivalent. In space, however, such a personal dosemeter has not yet been available and a dose equivalent is estimated through elaborate work using etched track detectors. It would be advantageous if we could simply measure the personal dose of an astronaut using conventional integrating dosemeters only. This possibility is discussed based on the concept of efficiency combination. PMID- 16709706 TI - Application of advanced Monte Carlo Methods in numerical dosimetry. AB - Many tasks in different sectors of dosimetry are very complex and highly sensitive to changes in the radiation field. Often, only the simulation of radiation transport is capable of describing the radiation field completely. Down to sub-cellular dimensions the energy deposition by cascades of secondary electrons is the main pathway for damage induction in matter. A large number of interactions take place until such electrons are slowed down to thermal energies. Also for some problems of photon transport a large number of photon histories need to be processed. Thus the efficient non-analogue Monte Carlo program, AMOS, has been developed for photon and electron transport. Various applications and benchmarks are presented showing its ability. For radiotherapy purposes the radiation field of a brachytherapy source is calculated according to the American Association of Physicists in Medicine Task Group Report 43 (AAPM/TG43). As additional examples, results for the detector efficiency of a high-purity germanium (HPGe) detector and a dose estimation for an X-ray shielding for radiation protection are shown. PMID- 16709707 TI - Neutron dosimetry with 6LiF-rich TL sheet. AB - A (6)LiF-rich thermoluminescent sheet-type dosemeter ((6)LiF-rich NTL sheet) was developed for neutron 2D dosimetry. The dosemeter utilises the (6)Li(n, alpha)(3)H reaction to detect thermal neutrons. Responses of the (6)LiF-rich NTL sheet to neutrons were measured at the neutron beam irradiation facility for BNCT in JRR-4 Research Reactor at the Japan Atomic Energy Research Institute. Placement of a multi-leaf collimator at the output port of the neutron (beam) irradiation facility, produced either stripe- or round-shaped neutron distributions; the spatial distribution was measured using the developed NTL sheets. Direct measurements of neutron attenuation in water were also carried out using the developed NTL sheet, submersed in a water phantom. In each experiment, NTL sheets having natural abundance (7.9%) of LiF, and (6)LiF-enriched NTL (18.94%) sheet were irradiated under the same conditions. The ratio of thermoluminescence intensities of the (6)LiF-rich NTL sheet to that of the normal NTL sheet was compared to a theoretically calculated value. The experimental measurements are shown to be in good agreement with the calculations. PMID- 16709708 TI - Defect centres and thermoluminescence in CaSO4:Dy,Ag phosphor. AB - The defect centres formed in the TL phosphor CaSO4:Dy,Ag are studied using the technique of Electron Spin Resonance. The Ag co-doped phosphor exhibits three glow peaks around 130, 220 and 375 degrees C in contrast with the two glow peaks observed in the CaSO4:Dy phosphor at 130 and 220 degrees C at a gamma ray dose of 1Gy. ESR studies show that the additional peak at 375 degrees C correlates with a Ag2+ centre formed due to gamma irradiation and observable only below -170 degrees C. The Ag2+ centre is characterised by an axial g-tensor with principal values g(parallel) = 2.38 and g(perpendicular) = 2.41. ESR studies further indicate that the precursor to a centre observable at low temperature (-170 degrees C) appears to act as the recombination centre for the TL peak at 375 degrees C; this radical is characterised by the g-values g(parallel) = 2.0023 and g(perpendicular) = 2.0038 and is assigned to SO3- radical. It is observed that there is more incorporation of Ag in the CaSO4:Dy system as compared with that in pure CaSO4 system. PMID- 16709709 TI - Efficiency-corrected dose verification with thermoluminescence dosemeters in heavy-ion beams. AB - One of the most essential difficulties in heavy-ion dosimetry by means of thermoluminescence dosemeters (TLDs)--often seen as a serious disadvantage of TLD utilisation--regards the changing TL-efficiency with increasing linear energy transfer (LET) of the particle. This behaviour leads to a falsification of absorbed dose that can be significant for many applications, e.g. in space or radiotherapeutic dosimetry. The high-temperature TL emission of LiF:Mg,Ti TL detectors can be exploited to obtain information about the LET of the heavy-ion radiation field under study. The high-temperature ratio (HTR) is used as a parameter to determine average LET. To correct the absorbed dose according to the TL-efficiency, the detailed dependence of HTR- and TL-efficiency on LET was recorded. These investigations were accomplished at the Heavy Ion Medical Accelerator (HIMAC) in Chiba, Japan, with a variety of high-energy ion beams (helium, carbon, neon, silicon and iron) ranging in LET from 2.2 to 393 keV microm(-1). The obtained relationships HTR vs. LET and TL-efficiency vs. LET were combined into a TL efficieny vs. HTR relationship. This enables correction of the absorbed dose (HTR-B method). The methodology is demonstrated by means of TLD 700 ((7)LiF:Mg,Ti) measurements in carbon beams of 290 and 400 MeV n(-1) available from HIMAC. PMID- 16709710 TI - Inter-comparison among different TLD-based techniques in a standard multisphere assembly for the characterisation of neutron fields. AB - In the framework of collaboration among the ENEA Radiation Protection Institute (Bologna), the ENEA Fusion Department (Frascati) and the INFN-LNF-Radiation Protection Group (Frascati), an experimental campaign was organised on the usage of thermoluminescence dosemeters (TLDs) for the dosimetric and spectrometric characterisation of neutron fields. Commercially available TLDs of different material and different sensitivity to photons and thermal neutrons were selected, namely TLD600H and TLD700H from Harshaw, GR206 and GR207 from SSDML (China), MCP 6s from TLD Poland. The detectors were first calibrated in standard fields of photons ((60)Co) and thermal neutrons at the ENEA-IRP Secondary Standard Calibration Laboratory of Bologna, then exposed in fast neutron standard fields of different energy, using a standard multisphere moderating assembly. The paper compares the dosimetric characteristics of the studied TL detectors, underlining the (n-gamma) discrimination capability, and discusses their spectrometric performances addressed to radiation protection applications. PMID- 16709711 TI - Commercial optical fibre as TLD material. AB - This work presents a study of commercial SiO2 optical fibre thermoluminescence (TL) properties as part of the efforts within the Dosimetric Application Project at the Physics Institute of the University of Mexico to develop new radiation detection materials and technologies. The SiO2 commercial optical fibre studied demonstrates useful TL properties and is an excellent candidate for use in TL dosimetry of ionising radiation. The optical fibre's glow curve was observed between 30 and 400 degrees C after exposure to 60Co gamma radiation. One very well-defined glow peak has a maximum at 230 degrees C. The TL response between 100 and 350 degrees C increases monotonically over a wide dose range, from 0.1 Gy to several kGy. It is linear in the range 0.1-3 Gy, which is important for clinical high dose or accident dosimetry. The optical fibre demonstrated high data reproducibility, low residual signal and almost no fading in our study. Moreover, the optical fibre can be re-used several times, after thermal annealing, without any detriment in the dose-response. All these TL characteristics, plus the small size of the 150 microm diameter SiO2 optical fibre, the high flexibility, easy handling and low cost compared with other TL materials, make the commercial optical fibre a very promising TL material for use in research, medicine, industry, reactors, and a variety of other applications. PMID- 16709712 TI - Comparative study of LiF:Mg,Cu,Na,Si and Li2B4O7:Cu,Ag,P TL detectors. AB - Recently, two new types of 'tissue equivalent' thermoluminescent detectors (TLDs) have aroused attention: LiF:Mg,Cu,Na,Si and Li2B4O7:Cu,Ag,P. In this work the characteristics of both detectors were compared with the characteristics of the well-known type LiF:Mg,Ti detector, TLD-100. The following properties were investigated: the glow curve structures, relative sensitivity, batch homogeneity and uniformity, detection threshold, reproducibility of the response, linearity in the wide dose range and fading. Also, the energy dependence for medium and low energy X rays was determined in the range of mean energies between 33 and 116 keV. The results confirmed 'tissue equivalency' of both new types in the investigated range of photon energies. LiF:Mg,Cu,Na,Si detector has very high sensitivity (approximately 75 times higher than that of TLD-100) and is convenient for use in a very low range of doses. Li2B4O7:Cu,Ag,P detector shows some improvements in comparison with the previously prepared types of lithium borate. The most important is the five times higher sensitivity than that of TLD 100. This detector is also very promising, especially in medical dosimetry. PMID- 16709713 TI - Evolution of the trapped charge distribution due to trap emptying processes in a natural aluminosilicate. AB - The evolution of the thermoluminescence glow curve of a natural Ca-Be rich aluminosilicate after annealing treatments at different temperatures has been studied in order to evaluate the changes in the trapped charge distribution. The glow curve consists of a single broad peak that continuously shifts toward higher temperatures when the sample is preheated up to increasing temperatures, thus indicating the presence of a continuous trap distribution. The glow curve fitting assuming different distribution functions shows how a gaussian distribution becomes a nearly exponential distribution owing to the thermal leakage of charge carriers from trapping centres. PMID- 16709714 TI - A comparison of quartz OSL and isothermal TL measurements on Chinese loess. AB - The reliability of equivalent doses (De) from Chinese loess, measured using isothermal thermoluminescence (ITL) is tested. Dose calculations use the single aliquot regenerative-dose (SAR) procedure. Despite good reproducibility of laboratory-induced signals and negligible response at zero dose, a significant overestimation of De is observed, compared with OSL measurements. Measurement of a known laboratory dose administered after optical bleaching, but before any heating, demonstrates that the first heating during measurement of the natural signal causes a significant sensitivity change, undetected by SAR. Using the single-aliquot regeneration and added (SARA) dose procedure, which allows for initial sensitivity change, good agreement with OSL is obtained after allowance is made for initial incomplete bleaching. It is concluded that SAR-ITL, in its present form, is not a suitable method for dating Chinese loess; it is very important to undertake a dose recovery test before any TL procedure is used to date sediments. PMID- 16709715 TI - Determination of quality parameters from statistical analysis of routine TLD dosimetry data. AB - Following the as low as reasonably achievable (ALARA) practice, there is a need to measure very low doses, of the same order of magnitude as the natural background, and the limits of detection of the dosimetry systems. The different contributions of the background signals to the total zero dose reading of thermoluminescence dosemeter (TLD) cards were analysed by using the common basic definitions of statistical indicators: the critical level (L(C)), the detection limit (L(D)) and the determination limit (L(Q)). These key statistical parameters for the system operated at NRC-Negev were quantified, based on the history of readings of the calibration cards in use. The electronic noise seems to play a minor role, but the reading of the Teflon coating (without the presence of a TLD crystal) gave a significant contribution. PMID- 16709716 TI - Study of gel materials as radioactive 222Rn gas detectors. AB - Commercial hair gel material (polyvinyl pyrolydone triethanolamine carbopol in water) and bacteriological agar (phycocolloid extracted from a group of red purple algae, usually Gelidium sp.) have been studied as radioactive radon gas detectors. The detection method is based on the diffusion of the radioactive gas in the gel material, and the subsequent measurement of trapped products of the natural decay of radon by gamma spectrometry. From the several radon daughters with gamma radiation emission (214Pb, 214Bi, 214Po, 210Pb, 210Po), two elements, 214Pb (0.352 MeV) and 214Bi (0.609 MeV), were chosen for the analysis in this work; in order to determine the best sensitivity, corrections were made for the short half-life of the analysed isotopes. For the gamma spectrometry analysis, a hyperpure germanium solid state detector was used, associated with a PC multichannel analyser card with Maestro and Microsoft Excel software. The results show the viability of the method: a linear response in a wide radon concentration range (450-10,000 Bq m(-3)), reproducibility of data, easy handling and low cost of the gel material. This detection methodology opens new possibilities for measurements of radon and other radioactive gases. PMID- 16709717 TI - On-board TLD measurements on MIR and ISS. AB - This paper presents results from dosimetric measurements made aboard the Mir space station and the International Space Station (ISS) using the Pille portable thermoluminescent dosemeter (TLD) system. This paper includes the dosimetry mapping and automatic readout (trapped and untrapped components) results from Mir and ISS. The mean dose rate in 2001-2003 was 7 microGy h(-1). Using the hourly measuring period in automatic mode, doses from both galactic (independent of South Atlantic Anomaly--SAA) and SAA components were determined during Euromir'95 experiment. The mean total dose rate was 12.5 microGy h(-1), while the SAA contribution was 6.2 microGy h(-1). A similar measurement was performed on ISS in 2001 and in 2003. Both the manual and automatic measurements show a significant decrease in dose rate in 2001 in comparison to 1995-1997 due to the change in solar activity. For determination of the high linear energy transfer contribution from the radiation field during the ISS mapping experiment, three CR-39 plastic nuclear track detectors (PNTDs) were co-located with each TL detector. Analysis of the combined TLD and PNTD measurements showed a typical mean TLD efficiency of 84%, a dose contribution <10 keV microm(-1) of 17%, and an average quality factor of 1.95. PMID- 16709718 TI - Comparison on characteristics of radiophotoluminescent glass dosemeters and thermoluminescent dosemeters. AB - The radiophotoluminescent glass dosemeter (RPLGD) system is applicable for measurement of radiation dose of X rays and gamma rays by using radiophotoluminescent glass (silver-activated phosphate glass). When the radiophotoluminescent glass is exposed to ionizing radiation, stable luminescent centres are created. During pulsed ultraviolet laser excitation (337.1 nm) in the reader, the centres emit a radiation induced orange fluorescent light (600-700 nm). This phenomenon is called radiophotoluminescence. This study compared the RPLGD system with lithium fluoride (LiF) thermoluminescence dosimetry system and the results of the study revealed that the RPLGD had not only good basic characteristics for reproducibility of readout value, dose linearity, energy dependence and fading, but also infinite repeatable measurements and could be one of the most important radiation dose measurement instruments. PMID- 16709719 TI - Effect of texture of plastic and elastic model foods on the parameters of mastication. AB - Mastication is continually modified throughout the chewing sequence in response to the texture of the food. The aim of this work was to compare the effects of an increase in hardness of two model food types, presenting either elastic or plastic rheological properties, on mastication. Each model food type consisted of four products of different hardness. Sensory testing experiments conducted with one group of 14 subjects showed significant perceived differences between products in terms of their increasing hardness. Fifteen other volunteers were asked to chew three replicates of each elastic and plastic product during two sessions. EMGs of masseter and temporalis muscles were recorded simultaneously with jaw movement during chewing. Numerous variables were analyzed from these masticatory recordings. Multiple linear regression analyses were used to assess the respective effects of food hardness and rheological properties on variables characterizing either the whole masticatory sequence or different stages of the sequence. Muscle activities were significantly affected by an increase in hardness regardless of the food type, whereas the shape of the cycles depended on the rheological properties. The masticatory frequency was affected by hardness at the initial stage of the sequence but overall frequency adaptation was better explained by a change in rheological behavior, with plastic products being chewed at a slower frequency. A dual hypothesis was proposed, implicating first a cortical-brain stem preprogrammed mechanism to adapt the shape of the jaw movements to the rheological properties of the food, and second, a brain stem mechanism with mainly sensory feedback from the mouth to adapt muscle force to the food hardness. PMID- 16709720 TI - Long-term potentiation is mediated by multiple kinase cascades involving CaMKII or either PKA or p42/44 MAPK in the adult rat dentate gyrus in vitro. AB - The induction of NMDA-receptor-dependent long-term potentiation (LTP) in adult CA1 is contingent on activation of Ca/calmodulin-dependent protein kinase II (CaMKII). However, little is known about kinase mediation of LTP in the dentate gyrus. In the present study, the involvement of the kinases CaMKII, PKA, and MAPK in the induction of LTP was studied in the dentate gyrus of adult rats. Individual application of selective inhibitors of CaMKII, MEK, or PKA did not inhibit induction of LTP. In contrast, coapplication of a CaMKII inhibitor with either a PKA or MEK inhibitor resulted in a strong block of LTP. Induction of LTP was blocked by the coapplication of the inhibitors CaMKII and PKA or MEK, both when they were applied 1 h before the induction stimulus and also when they were applied after the induction stimulus. Thus LTP is mediated by either of two parallel cascades, one involving CaMKII and the other PKA or MAPK. Moreover, these cascades are active for a certain period after the induction stimulus. PMID- 16709721 TI - Role of the Na+-K+-2Cl- cotransporter in the development of capsaicin-induced neurogenic inflammation. AB - Recent behavioral and electrophysiological studies have attributed an important role to dorsal root reflexes (DRRs) in the initiation and development of neurogenic inflammation produced by intradermal capsaicin (CAP). The DRRs can occur in peptidergic fibers, resulting in peripheral release of neuromediators that produce vasodilation, plasma extravasation and subsequently hyperalgesia and allodynia. In this study, we have evaluated the effect of spinal administration of bumetanide (a blocker of the Na+-K+-2Cl- cotransporter, NKCC) on DRR activity, changes in cutaneous blood flow (vasodilation), hindpaw edema, mechanical allodynia, and hyperalgesia induced by intradermal injection of 1% CAP in Sprague Dawley rats. Vasodilation was monitored using laser Doppler flowmetry, neurogenic edema was evaluated by measurements of hindpaw volume, and secondary mechanical allodynia and hyperalesia were tested using von Frey filaments (10 and 200 mN) applied to the plantar surface of the paw. Changes in the blood flow were blocked significantly by intrathecal bumetanide at 10 and 100 microM in both pre- and posttreatment studies. Spinal bumetanide at 10 and 100 microM blocked neurogenic edema when it was administered before CAP injection, but only bumetanide at 100 microM administered after CAP injection reduced the paw edema significantly. Furthermore, the administration of bumetanide onto the spinal cord reduced the increment in DRR activity produced by CAP. Finally, both secondary mechanical allodynia and hyperalesia were reduced by bumetanide at 1, 10, and 100 microM. Taken together these results suggest that NKCC is involved in the increases in DRR activity, neurogenic inflammation and hyperalgesia and allodynia induced by intradermal CAP. PMID- 16709722 TI - Sound localization by barn owls in a simulated echoic environment. AB - We examined the accuracy and precision with which the barn owl (Tyto alba) turns its head toward sound sources under conditions that evoke the precedence effect (PE) in humans. Stimuli consisted of 25-ms noise bursts emitted from two sources, separated horizontally by 40 degrees, and temporally by 3-50 ms. At delays from 3 to 10 ms, head turns were always directed at the leading source, and were nearly as accurate and precise as turns toward single sources, indicating that the leading source dominates perception. This lead dominance is particularly remarkable, first, because on some trials, the lagging source was significantly higher in amplitude than the lead, arising from the directionality of the owl's ears, and second, because the temporal overlap of the two sounds can degrade the binaural cues with which the owl localizes sounds. With increasing delays, the influence of the lagging source became apparent as the head saccades became increasingly biased toward the lagging source. Furthermore, on some of the trials at delays > or = 20 ms, the owl turned its head, first, in the direction of one source, and then the other, suggesting that it was able to resolve two separately localizable sources. At all delays <50 ms, response latencies were longer for paired sources than for single sources. With the possible exception of response latency, these findings demonstrate that the owl exhibits precedence phenomena in sound localization similar to those in humans and cats, and provide a basis for comparison with neurophysiological data. PMID- 16709723 TI - Presynaptic GABA(B) receptors regulate retinohypothalamic tract synaptic transmission by inhibiting voltage-gated Ca2+ channels. AB - Presynaptic GABA(B) receptor activation inhibits glutamate release from retinohypothalamic tract (RHT) terminals in the suprachiasmatic nucleus (SCN). Voltage-clamp whole cell recordings from rat SCN neurons and optical recordings of Ca2+-sensitive fluorescent probes within RHT terminals were used to examine GABA(B)-receptor modulation of RHT transmission. Baclofen inhibited evoked excitatory postsynaptic currents (EPSCs) in a concentration-dependent manner equally during the day and night. Blockers of N-, P/Q-, T-, and R-type voltage dependent Ca2+ channels, but not L-type, reduced the EPSC amplitude by 66, 36, 32, and 18% of control, respectively. Joint application of multiple Ca2+ channel blockers inhibited the EPSCs less than that predicted, consistent with a model in which multiple Ca2+ channels overlap in the regulation of transmitter release. Presynaptic inhibition of EPSCs by baclofen was occluded by omega-conotoxin GVIA (< or = 72%), mibefradil (< or = 52%), and omega-agatoxin TK (< or = 15%), but not by SNX-482 or nimodipine. Baclofen reduced both evoked presynaptic Ca2+ influx and resting Ca2+ concentration in RHT terminals. Tertiapin did not alter the evoked EPSC and baclofen-induced inhibition, indicating that baclofen does not inhibit glutamate release by activation of Kir3 channels. Neither Ba2+ nor high extracellular K+ modified the baclofen-induced inhibition. 4-Aminopyridine (4-AP) significantly increased the EPSC amplitude and the charge transfer, and dramatically reduced the baclofen effect. These data indicate that baclofen inhibits glutamate release from RHT terminals by blocking N-, T-, and P/Q-type Ca2+ channels, and possibly by activation of 4-AP-sensitive K+ channels, but not by inhibition of R- and L-type Ca2+ channels or by Kir3 channel activation. PMID- 16709726 TI - Powerful motion illusion caused by temporal asymmetries in ON and OFF visual pathways. AB - Successive presentations of Glass patterns (randomly positioned pairs of dots oriented in a coherent pattern) create a strong sense of global motion along the orientation of the pattern, but ambiguous in direction. Here we report that dynamic "anti-Glass" patterns, created by successive pairs of globally structured pairs of opposite polarity, create an even more powerful motion illusion that is unambiguous in direction: the dark dots always move toward the light. The motion can be cancelled and reversed by introducing a real delay in the presentation of the light dots, suggesting that the effective stimulation of the light is about 3 ms faster than the dark dots. The most plausible explanation for this is that human on channels are faster than off channels, as has been shown in the macaque. PMID- 16709725 TI - Afferent encoding of central oscillations in the monkey arm. AB - We have investigated whether peripheral afferent fibers could encode the central oscillations that are commonly seen in the primate motor system. We analyzed 52 single afferent recordings from the C8/T1 dorsal root ganglia of two monkeys performing an isometric wrist flexion-extension task. Coherence and directed coherence were calculated between the afferent spikes and forearm EMG. Seven of 52 cells were identified as Group Ia afferents by the production of narrow postspike facilitation in spike-triggered averages of rectified EMG. These identified afferents showed significant coherence, and directed coherence, with EMG over a wide frequency range. By contrast, coherence was weak for a population that showed little directional preference for flexion or extension movements during task performance, and probably contained mainly cutaneous afferents. Oscillations are known to appear in muscle activity; their presence in afferent firing as well implies that central oscillations pass around a peripheral feedback loop and may be involved in sensorimotor integration. PMID- 16709724 TI - Signal transmission between gap-junctionally coupled passive cables is most effective at an optimal diameter. AB - We analyze simple morphological configurations that represent gap-junctional coupling between neuronal processes or between muscle fibers. Specifically, we use cable theory and simulations to examine the consequences of current flow from one cable to other gap-junctionally coupled passive cables. When the proximal end of the first cable is voltage clamped, the amplitude of the electrical signal in distal portions of the second cable depends on the cable diameter. However, this amplitude does not simply increase if cable diameter is increased, as expected from the larger length constant; instead, an optimal diameter exists. The optimal diameter arises because the dependency of voltage attenuation along the second cable on cable diameter follows two opposing rules. As cable diameter increases, the attenuation decreases because of a larger length constant yet increases because of a reduction in current density arising from the limiting effect of the gap junction on current flow into the second cable. The optimal diameter depends on the gap junction resistance and cable parameters. In branched cables, dependency on diameter is local and thus may serve to functionally compartmentalize branches that are coupled to other cells. Such compartmentalization may be important when periodic signals or action potentials cause the current flow across gap junctions. PMID- 16709727 TI - Breast cancer resistance protein (BCRP/ABCG2) is expressed by progenitor cells/reactive ductules and hepatocytes and its expression pattern is influenced by disease etiology and species type: possible functional consequences. AB - Breast cancer resistance protein (BCRP/ABCG2) is an ATP-binding cassette transport protein that is expressed in several organs including the liver. Previous studies have shown that ABC transport proteins play an important pathophysiological role in several liver diseases. However, to date, expression pattern and possible role of BCRP in human liver diseases and animal models have not been studied in detail. Here we investigated the expression pattern of BCRP in normal liver, chronic parenchymal and biliary human liver diseases, and parallel in different rat models of liver diseases. Expression was studied by immunohistochemistry and additionally by RT-PCR analysis in Thy-1-positive rat oval cells. Bile ducts, hepatic progenitor cells, reactive bile ductules, and blood vessel endothelium were immunoreactive for BCRP in normal liver and all types of human liver diseases and in rat models. BCRP was expressed by the canalicular membrane of hepatocytes in normal and diseased human liver, but never in rat liver. Remarkably, there was also expression of BCRP at the basolateral pole of human hepatocytes, and this was most pronounced in chronic biliary diseases. In conclusion, BCRP positivity in the progenitor cells/reactive ductules could contribute to the resistance of these cells to cytotoxic agents and xenotoxins. Basolateral hepatocytic expression in chronic biliary diseases may be an adaptive mechanism to pump bile constituents back into the sinusoidal blood. Strong differences between human and rat liver must be taken into account in future studies with animal models. PMID- 16709728 TI - A novel fluorescent probe that is brain permeable and selectively binds to myelin. AB - Myelin is a multilayered glial cell membrane that forms segmented sheaths around large-caliber axons of both the central nervous system (CNS) and peripheral nervous system (PNS). Myelin covering insures rapid and efficient transmission of nerve impulses. Direct visual assessment of local changes of myelin content in vivo could greatly facilitate diagnosis and therapeutic treatments of myelin related diseases. Current histologic probes for the visualization of myelin are based on antibodies or charged histochemical reagents that do not enter the brain. We have developed a series of chemical compounds including (E,E)-1,4 bis(4'-aminostyryl)-2-dimethoxy-benzene termed BDB and the subject of this report, which readily penetrates the blood-brain barrier and selectively binds to the myelin sheath in brain. BDB selectively stains intact myelinated regions in wild-type mouse brain, which allows for delineation of cuprizone-induced demyelinating lesions in mouse brain. BDB can be injected IV into the brain and selectively detect demyelinating lesions in cuprizone-treated mice in situ. These studies justified further investigation of BDB as a potential myelin-imaging probe to monitor myelin pathology in vivo. PMID- 16709729 TI - Protease analysis by neoepitope approach reveals the activation of MMP-9 is achieved proteolytically in a test tissue cartilage model involved in bone formation. AB - A principle of regulation of matrix metalloproteinase (MMP) activity has been introduced as the cysteine-switch mechanism of activation (Springman et al. 1990). According to this mechanism, a critical Cys residue found in the auto inhibitory propeptide domain of latent proenzyme is important to determine whether or not activation is turned on or off. The mechanism further allows for multiple modes of activation. To determine whether or not activation is accomplished proteolytically within a rat test cartilage model, protease analysis by the neoepitope approach, which relies upon a set of antibodies, was applied. One is used to identify the MMP-9 proenzyme bearing the critical cysteine residue, the other to identify any enzyme present bearing a new NH2-terminus 89FQTFD. This is indicative of MMP-9 lacking the cysteine switch. The antibody set has been applied to frozen tissue sections and analyzed by light and electron microscopic methods. Results reveal that activation of the MMP-9 protease involves limited proteolysis resulting in propeptide domain release. Here we report the observed changes of protease form to indigenous cells and extracellular matrix, thereby making it possible to uncover the features of MMP-9 activation within a specified set of tissue circumstances where a cartilage model is transformed into definitive bone. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials. PMID- 16709730 TI - Expression of the human copper influx transporter 1 in normal and malignant human tissues. AB - The major copper influx transporter, copper transporter 1 (hCTR1), controls the cellular accumulation of cisplatin in mammalian cells. The goal of this study was to determine the pattern of hCTR1 expression in normal and malignant human tissues. Tissue arrays were stained with an antibody specific for hCTR1 using standard immunohistochemical techniques. Particularly strong staining was noted in the alpha cells of the pancreatic islets, enteroendocrine cells of the gastric mucosa and bronchioles, C cells of the thyroid, and a subset of cells in the anterior pituitary. Frequency and intensity of hCTR1 staining in malignant tissues reflected the levels found in their normal tissue counterparts. For example, neither normal prostate nor prostate cancers expressed hCTR1, whereas it was commonly expressed in both normal colonic epithelium and in colon carcinomas. Strong staining was observed in a limited number of cases of carcinoid tumors, Ewing's sarcoma, and undifferentiated carcinomas. Although all tissues require copper, expression of hCTR1 was highly variable among normal tissues and among the major human malignancies, with the highest levels found in enteroendocrine cells. No hCTR1 expression was found in several common types of cancer, suggesting that hCTR1 expression is not commonly enhanced by transformation. PMID- 16709731 TI - Histochemical and electron microscopic analysis of spiculogenesis in the demosponge Suberites domuncula. AB - The skeleton of demosponges is built of spicules consisting of biosilica. Using the primmorph system from Suberites domuncula, we demonstrate that silicatein, the biosilica-synthesizing enzyme, and silicase, the catabolic enzyme, are colocalized at the surface of growing spicules as well as in the axial filament located in the axial canal. It is assumed that these two enzymes are responsible for the deposition of biosilica. In search of additional potential structural molecules that might guide the mineralization process during spiculogenesis to species-specific spicules, electron microscopic studies with antibodies against galectin and silicatein were performed. These studies showed that silicatein forms a complex with galectin; the strings/bundles of this complex are intimately associated with the surface of the spicules and arranged concentrically around them. Collagen fibers are near the silactein/galectin complexes. The strings/bundles formed from silicatein/galectin display a lower degree of orientation than the collagen fibers arranged in a highly ordered pattern around the spicules. These data indicate that species-specific formation of spicules involves a network of (diffusible) regulatory factor(s) controlling enzymatic silica deposition; this mineralization process proceeds on a galectin/collagen organic matrix. PMID- 16709733 TI - Angelo Mosso and muscular fatigue: 116 years after the first Congress of Physiologists: IUPS commemoration. AB - At the first International Congress of Physiologists in Basel, Switzerland, the Italian physiologist Angelo Mosso (1846-1910) discussed his findings on muscular fatigue while demonstrating the functioning of an ergograph (work recorder). One hundred sixteen years later, Mosso's career, scientific accomplishments, and legacy in the study of muscular fatigue were commemorated at the 2005 International Congress of Physiological Sciences. After receiving his degree in Medicine and Surgery from Turin, Italy, in 1870, Mosso was able to study and interact with renowned physiologists as Wilhelm Ludwig, Du Bois-Reymond, Hugo Kronecker, and Etienne Marey. By 1879, he was Professor of Physiology at the University in Turin, where he conducted research pertaining to blood circulation, respiration, physical education, high-altitude physiology, and muscular fatigue. Using tracings from the ergograph (concentric contractions of the flexor muscles of the middle finger that were volitionally or electrically stimulated), he was able to characterize muscle fatigue and to associate its occurrence with central or peripheral influences. He demonstrated that exercise would increase muscular strength and endurance while prolonging the occurrence of fatigue, which he postulated was a chemical process that involved the production of toxic substances such as carbonic acid. The phenomenon of contracture was described, and his collective studies led to the formulation of laws pertaining to exhaustion and to the 1891 publication of La Fatica (Fatigue). Besides La Fatica, Mosso will be remembered as a scientist with a love for physiology, a concern for the social welfare of his countrymen, and as one who sought to integrate physiological, philosophical, and psychological concepts in his experimental studies. PMID- 16709732 TI - Predicting RNA pseudoknot folding thermodynamics. AB - Based on the experimentally determined atomic coordinates for RNA helices and the self-avoiding walks of the P (phosphate) and C4 (carbon) atoms in the diamond lattice for the polynucleotide loop conformations, we derive a set of conformational entropy parameters for RNA pseudoknots. Based on the entropy parameters, we develop a folding thermodynamics model that enables us to compute the sequence-specific RNA pseudoknot folding free energy landscape and thermodynamics. The model is validated through extensive experimental tests both for the native structures and for the folding thermodynamics. The model predicts strong sequence-dependent helix-loop competitions in the pseudoknot stability and the resultant conformational switches between different hairpin and pseudoknot structures. For instance, for the pseudoknot domain of human telomerase RNA, a native-like and a misfolded hairpin intermediates are found to coexist on the (equilibrium) folding pathways, and the interplay between the stabilities of these intermediates causes the conformational switch that may underlie a human telomerase disease. PMID- 16709734 TI - A classic learning opportunity from Fenn, Rahn, and Otis (1946): the alveolar gas equation. AB - The alveolar gas equation, the focus of a classic paper by Fenn, Rahn, and Otis, provides a framework for understanding the mechanisms involved in pulmonary gas exchange as well as the limits of human performance. The classic 1946 paper by Fehn, Rahn, and Otis gives your students an opportunity to learn about the alveolar gas equation from the physiologists who pioneered it and demonstrates that mathematics and data graphics are fundamental tools with which to learn respiratory physiology. In this essay, I outline avenues of discovery by which your students can explore the alveolar gas equation. Meaningful learning stems from inspiration: to learn, you must be inspired to learn. If anyone can inspire learning in respiratory physiology, it is Wallace Fenn, Hermann Rahn, and Arthur Otis. PMID- 16709735 TI - Using a classic paper by Gottschalk and Mylle to teach the countercurrent model of urinary concentration. AB - Most undergraduates lack the scientific background to read and appreciate much of the primary literature in physiology. Even when the underlying concepts are elegantly simple, the inherent complexity of contemporary papers often makes the work inaccessible to them. However, with a little help, they can be guided to an understanding of the creative thought processes that underlie the research and to appreciate its significance. This is especially true of many classic papers in physiology that often rely on easily comprehensible techniques. Moreover, the American Physiological Society (APS) has invited prominent scientists to select important papers in their fields and to write essays that both put the work into historical context and explain why it is scientifically important. The APS Legacy Project makes these classic papers freely available online. One such paper by Gottschalk and Mylle presents data from a series of micropuncture studies that confirm all of the predictions of the countercurrent exchange model of concentrated urine production (2). The included handout of questions for discovery learning and teaching points suggest ways to use the paper as an instructional resource. PMID- 16709737 TI - The Opinion Editorial: teaching physiology outside the box. AB - Improving the public understanding of science is an important challenge for the future professional scientists who are our current undergraduates. In this paper, we present a conceptual model that explores the role of mass media as community gatekeepers of new scientific findings. This model frames the benefits for undergraduate science students to learn about media genres so that they can learn to communicate science more effectively to nonprofessional audiences. Informed by this Media Role model, we then detail a novel writing task for undergraduate physiology students, the Opinion Editorial (Op-Ed), and an accompanying Peer Review. The Op-Ed genre was directly taught to the students by a professional journalist. As an assessment task, students presented a recent, highly technical paper as an Op-Ed. This was assessed by both faculty members and peers using a detailed assessment rubric. Most students were able to replicate the features of Op-Eds and attained high grades on their writing tasks. Survey data from final year physiology students (n = 230) were collected before and after the implementation of the Op-Ed/Peer Review. These indicated that most students were aware of the importance of scientists to effectively communicate their knowledge to nonprofessional audiences, that the Op-Ed writing task was challenging, and that they believed that their ability to write to nonprofessional audiences was improved after explicit teaching and feedback. PMID- 16709736 TI - Teaching from classic papers: Hill's model of muscle contraction. AB - A. V. Hill's 1938 paper "The heat of shortening and the dynamic constants of muscle" is an enduring classic, presenting detailed methods, meticulous experiments, and the model of muscle contraction that now bears Hill's name. Pairing a simulation based on Hill's model with a reading of his paper allows students to follow his thought process to discover key principles of muscle physiology and gain insight into how to develop quantitative models of physiological processes. In this article, the experience of the author using this approach in a graduate biomedical engineering course is outlined, along with suggestions for adapting this approach to other audiences. PMID- 16709738 TI - A "how to" guide for developing a publishable Scholarship of Teaching project. AB - In order for the efforts of scholarly teaching to be recognized, the work must become public and presented to peers for review. Scholarship of Teaching is not only improving instruction and learning but also methodically assessing whether specific teaching interventions have had the desired effect. In this paper, the author presents a step-by-step guide for how to develop a Scholarship of Teaching project that is well thought out and worthy of publication. Factors to consider before performing such pedagogical research include developing a clear research hypothesis for the classroom, reviewing the background literature, obtaining Institutional Review Board (Human Subjects) approval, and determining which methods of assessment may be used. This "how to" guide discusses how to handle all of these factors and prepare the data for publication and introduces the reader to references related to the Scholarship of Teaching and learning as well as educational research and theory. PMID- 16709739 TI - Pizza and pasta help students learn metabolism. AB - In this article, we report on an experiment designed to improve the learning of metabolic biochemistry by nutrition and medical undergraduate students. Twelve students participated in a monitored lunch and had their blood extracted for analysis 1) before lunch, 2) 30 min after lunch, and 3) 3 h after lunch. The subjects were divided in two groups. One group had a hyperglicidic meal [pasta plus orange juice: 80% carbohydrate, 10% protein, and 10% lipid (estimated values)] and the other group had a hyperlipidic meal (calabresi pizza plus diet soda: 36% carbohydrate, 18% protein, and 46% lipid). Individual quantities of food were based on body mass index, age, and sex. The blood parameters analyzed were glucose, triglycerides (TG), and urea. Glucose remained constant in the three measurements in both groups. The TG concentration in the pasta group was constant before and after lunch but increased significantly during the evening. In the pizza group, TG increased after lunch and remained constant in the evening. Levels of urea increased only in the evening, specially in the pizza group. These results were used for the final biochemistry exam. With the maximum score set as 10, the average score was 6.0 +/- 2.4 (n = 102). We considered this activity a unique way of evaluating important issues on metabolism, because students had several hours to work on the final exam (with free access to a bibliography). It was also a good didactic experience (problem-based learning like) for the subject students, because they had to work in all phases of the experiment (idealization, realization, and analysis) and participated actively in the elaboration and correction of the exam. PMID- 16709740 TI - Medical physiology and experimentation: reconsidering the undergraduate examination structure. PMID- 16709742 TI - The blanket method: a novel method of teaching peritoneal relations of female reproductive organs. PMID- 16709743 TI - The rapid-response: a break during lecture. PMID- 16709746 TI - The structure of an infectious P22 virion shows the signal for headful DNA packaging. AB - Bacteriophages, herpesviruses, and other large double-stranded DNA (dsDNA) viruses contain molecular machines that pump DNA into preassembled procapsids, generating internal capsid pressures exceeding, by 10-fold, that of bottled champagne. A 17 angstrom resolution asymmetric reconstruction of the infectious P22 virion reveals that tightly spooled DNA about the portal dodecamer forces a conformation that is significantly different from that observed in isolated portals assembled from ectopically expressed protein. We propose that the tight dsDNA spooling activates the switch that signals the headful chromosome packing density to the particle exterior. PMID- 16709745 TI - Variable very-high-energy gamma-ray emission from the microquasar LS I +61 303. AB - Microquasars are binary star systems with relativistic radio-emitting jets. They are potential sources of cosmic rays and can be used to elucidate the physics of relativistic jets. We report the detection of variable gamma-ray emission above 100 gigaelectron volts from the microquasar LS I 61 + 303. Six orbital cycles were recorded. Several detections occur at a similar orbital phase, which suggests that the emission is periodic. The strongest gamma-ray emission is not observed when the two stars are closest to one another, implying a strong orbital modulation of the emission or absorption processes. PMID- 16709747 TI - Food-caching western scrub-jays keep track of who was watching when. AB - Western scrub-jays (Aphelocoma californica) hide food caches for future consumption, steal others' caches, and engage in tactics to minimize the chance that their own caches will be stolen. We show that scrub-jays remember which individual watched them during particular caching events and alter their recaching behavior accordingly. We found no evidence to suggest that a storer's use of cache protection tactics is cued by the observer's behavior. PMID- 16709749 TI - Science as smoke screen. PMID- 16709748 TI - Autoreactive B cell responses to RNA-related antigens due to TLR7 gene duplication. AB - Antibodies against nuclear self-antigens are characteristic of systemic autoimmunity, although mechanisms promoting their generation and selection are unclear. Here, we report that B cells containing the Y-linked autoimmune accelerator (Yaa) locus are intrinsically biased toward nucleolar antigens because of increased expression of TLR7, a single-stranded RNA-binding innate immune receptor. The TLR7 gene is duplicated in Yaa mice because of a 4-Megabase expansion of the pseudoautosomal region. These results reveal high divergence in mouse Y chromosomes and represent a good example of gene copy number qualitatively altering a polygenic disease manifestation. PMID- 16709750 TI - Korean cloning scandal. Prosecutors allege elaborate deception and missing funds. PMID- 16709751 TI - U.S. mathematics education. Well-balanced panel to tackle algebra reform. PMID- 16709752 TI - U.S. patent policy. PTO wants to tap experts to help patent examiners. PMID- 16709753 TI - Paleoanthropology. How the hobbit shrugged: tiny hominid's story takes new turn. PMID- 16709754 TI - Paleoanthropology. But is it pathological? PMID- 16709755 TI - Genetic testing. U.K. embryos may be screened for cancer risk. PMID- 16709756 TI - Human evolution. Genomes throw kinks in timing of chimp-human split. PMID- 16709758 TI - Scientific misconduct. Invention of China's homegrown DSP chip dismissed as a hoax. PMID- 16709757 TI - infectious disease. RU-486-linked deaths open debate about risky bacteria. PMID- 16709761 TI - Energy alternatives. Waiting for ITER, fusion jocks look EAST. PMID- 16709760 TI - Astronomy. After a tough year, ALMA's star begins to rise at last. PMID- 16709759 TI - Education. Finding common ground in the U.S. math wars. PMID- 16709762 TI - Energy alternatives. Asian fusion. PMID- 16709763 TI - Scientific openness. Should academics self-censor their findings on terrorism? PMID- 16709764 TI - Translation research and drug development. PMID- 16709765 TI - Extinct or possibly extinct? PMID- 16709766 TI - Incorporating evolution into medical education. PMID- 16709767 TI - Benzene exposure and hematotoxicity. PMID- 16709768 TI - Comment on "The Brain of LB1, Homo floresiensis". AB - Endocast analysis of the brain Homo floresiensis by Falk et al. (Reports, 8 April 2005, p. 242) implies that the hominid is an insular dwarf derived from H. erectus, but its tiny cranial capacity cannot result from normal dwarfing. Consideration of more appropriate microcephalic syndromes and specimens supports the hypothesis of modern human microcephaly. PMID- 16709769 TI - Environment. Investments toward sustainable development. PMID- 16709770 TI - Materials science. Making high-flux membranes with carbon nanotubes. PMID- 16709771 TI - Chemistry. Mode-selective control of surface reactions. PMID- 16709772 TI - Applied physics. Assembly and probing of spin chains of finite size. PMID- 16709773 TI - Behavior. Foresight and evolution of the human mind. PMID- 16709774 TI - Neuroscience. Gatekeeper at the synapse. PMID- 16709775 TI - Chemistry. Dangerously seeking linear carbon. PMID- 16709776 TI - Genomics and the irreducible nature of eukaryote cells. AB - Large-scale comparative genomics in harness with proteomics has substantiated fundamental features of eukaryote cellular evolution. The evolutionary trajectory of modern eukaryotes is distinct from that of prokaryotes. Data from many sources give no direct evidence that eukaryotes evolved by genome fusion between archaea and bacteria. Comparative genomics shows that, under certain ecological settings, sequence loss and cellular simplification are common modes of evolution. Subcellular architecture of eukaryote cells is in part a physical-chemical consequence of molecular crowding; subcellular compartmentation with specialized proteomes is required for the efficient functioning of proteins. PMID- 16709777 TI - Dispersal limitations matter for microbial morphospecies. AB - In contrast with macroorganisms, whose geographical ranges are typically restricted, many microbial species appear to have cosmopolitan distributions. This observation has been explained as a consequence of ubiquitous dispersal caused by the enormous population sizes of microbial species. Recently, this "everything is everywhere, but, the environment selects" theory has been challenged by the detection of considerable regional genetic variability within microbial morphospecies. We demonstrate that, contrary to what is expected under ubiquitous dispersal, evidence of regional-scale metacommunity processes can be detected in microbial morphospecies. Our results imply that the microbial and macrobial world are structured by analogous processes. PMID- 16709778 TI - Desorption of H from Si(111) by resonant excitation of the Si-H vibrational stretch mode. AB - Past efforts to achieve selective bond scission by vibrational excitation have been thwarted by energy thermalization. Here we report resonant photodesorption of hydrogen from a Si(111) surface using tunable infrared radiation. The wavelength dependence of the desorption yield peaks at 0.26 electron volt: the energy of the Si-H vibrational stretch mode. The desorption yield is quadratic in the infrared intensity. A strong H/D isotope effect rules out thermal desorption mechanisms, and electronic effects are not applicable in this low-energy regime. A molecular mechanism accounting for the desorption event remains elusive. PMID- 16709779 TI - Oligonucleotide-modified gold nanoparticles for intracellular gene regulation. AB - We describe the use of gold nanoparticle-oligonucleotide complexes as intracellular gene regulation agents for the control of protein expression in cells. These oligonucleotide-modified nanoparticles have affinity constants for complementary nucleic acids that are higher than their unmodified oligonucleotide counterparts, are less susceptible to degradation by nuclease activity, exhibit greater than 99% cellular uptake, can introduce oligonucleotides at a higher effective concentration than conventional transfection agents, and are nontoxic to the cells under the conditions studied. By chemically tailoring the density of DNA bound to the surface of gold nanoparticles, we demonstrated a tunable gene knockdown. PMID- 16709780 TI - Preparation of poly(diiododiacetylene), an ordered conjugated polymer of carbon and iodine. AB - Conjugated organic polymers generally must include large substituents for stability, either contained within or appended to the polymer chain. In polydiacetylenes, the substituents fulfill another important role: During topochemical polymerization, they control the spacing between the diyne monomers to produce an ordered polymer. By using a co-crystal scaffolding, we have prepared poly(diiododiacetylene), or PIDA, a nearly unadorned carbon chain substituted with only single-atom iodine side groups. The monomer, diiodobutadiyne, forms co-crystals with bis(nitrile) oxalamides, aligned by hydrogen bonds between oxalamide groups and weak Lewis acid-base interactions between nitriles and iodoalkynes. In co-crystals with one oxalamide host, the diyne undergoes spontaneous topochemical polymerization to form PIDA. The structure of the dark blue crystals, which look copper-colored under reflected light, has been confirmed by single-crystal x-ray diffraction, ultraviolet visible absorption spectroscopy, and scanning electron microscopy. PMID- 16709781 TI - Fast mass transport through sub-2-nanometer carbon nanotubes. AB - We report gas and water flow measurements through microfabricated membranes in which aligned carbon nanotubes with diameters of less than 2 nanometers serve as pores. The measured gas flow exceeds predictions of the Knudsen diffusion model by more than an order of magnitude. The measured water flow exceeds values calculated from continuum hydrodynamics models by more than three orders of magnitude and is comparable to flow rates extrapolated from molecular dynamics simulations. The gas and water permeabilities of these nanotube-based membranes are several orders of magnitude higher than those of commercial polycarbonate membranes, despite having pore sizes an order of magnitude smaller. These membranes enable fundamental studies of mass transport in confined environments, as well as more energy-efficient nanoscale filtration. PMID- 16709782 TI - Apes save tools for future use. AB - Planning for future needs, not just current ones, is one of the most formidable human cognitive achievements. Whether this skill is a uniquely human adaptation is a controversial issue. In a study we conducted, bonobos and orangutans selected, transported, and saved appropriate tools above baseline levels to use them 1 hour later (experiment 1). Experiment 2 extended these results to a 14 hour delay between collecting and using the tools. Experiment 3 showed that seeing the apparatus during tool selection was not necessary to succeed. These findings suggest that the precursor skills for planning for the future evolved in great apes before 14 million years ago, when all extant great ape species shared a common ancestor. PMID- 16709783 TI - Agent-specific responses in the cingulate cortex during economic exchanges. AB - Interactions with other responsive agents lie at the core of all social exchange. During a social exchange with a partner, one fundamental variable that must be computed correctly is who gets credit for a shared outcome; this assignment is crucial for deciding on an optimal level of cooperation that avoids simple exploitation. We carried out an iterated, two-person economic exchange and made simultaneous hemodynamic measurements from each player's brain. These joint measurements revealed agent-specific responses in the social domain ("me" and "not me") arranged in a systematic spatial pattern along the cingulate cortex. This systematic response pattern did not depend on metrical aspects of the exchange, and it disappeared completely in the absence of a responding partner. PMID- 16709785 TI - Biogeographic evolution of Madagascar's microendemic biota. AB - The endemic species richness on Madagascar, relative to landmass area, is unparalleled in the world. Many organisms on the island have restricted geographical ranges. A comprehensive hypothesis explaining the evolution of this microendemism has yet to be developed. Using an analysis of watersheds in the context of Quaternary climatic shifts, we provide a new mechanistic model to explain the process of explosive speciation on the island. River catchments with sources at relatively low elevations were zones of isolation and hence led to the speciation of locally endemic taxa, whereas those at higher elevations were zones of retreat and dispersion and hence contain proportionately lower levels of microendemism. These results provide a framework for biogeographic and phylogeographic studies, as well as a basis for prioritizing conservation actions of the remaining natural forest habitats on the island. PMID- 16709789 TI - Infants with perinatal hypoxic ischemia: feasibility of fiber tracking at birth and 3 months. AB - The purpose of this study was to retrospectively investigate the feasibility of fiber tracking at birth and 3 months in infants with hypoxic ischemia to detect disturbances in white matter development. This retrospective study did not require institutional review board approval. All parents gave informed consent. Diffusion-tensor magnetic resonance (MR) images were obtained in full-term newborns: seven with standard MR imaging findings and 10 with perinatal hypoxic ischemia-related abnormalities. Visualization of white matter tracts was investigated by using a volume-tracing and quantification technique. Fiber tracking was useful for studying the neonatal brain. Abnormalities resulted in fiber patterns that were different from the fiber patterns of normal brain tissue. The corona radiata fibers were frequently affected. PMID- 16709784 TI - A systems approach to mapping DNA damage response pathways. AB - Failure of cells to respond to DNA damage is a primary event associated with mutagenesis and environmental toxicity. To map the transcriptional network controlling the damage response, we measured genomewide binding locations for 30 damage-related transcription factors (TFs) after exposure of yeast to methyl methanesulfonate (MMS). The resulting 5272 TF-target interactions revealed extensive changes in the pattern of promoter binding and identified damage specific binding motifs. As systematic functional validation, we identified interactions for which the target changed expression in wild-type cells in response to MMS but was nonresponsive in cells lacking the TF. Validated interactions were assembled into causal pathway models that provide global hypotheses of how signaling, transcription, and phenotype are integrated after damage. PMID- 16709790 TI - Indirect MR arthrography of the shoulder: use of abduction and external rotation to detect full- and partial-thickness tears of the supraspinatus tendon. AB - PURPOSE: To evaluate prospectively the accuracy of indirect magnetic resonance (MR) arthrography for supraspinatus tendon tears during neutral positioning or abduction and external rotation (ABER) and neutral positioning. MATERIALS AND METHODS: Informed consent was obtained in all patients, and the study was approved by the institutional review board. Indirect MR arthrography of the shoulder was performed in 51 symptomatic patients (14 female, 37 male; mean age, 47 years) in the neutral position (set 1) and in the neutral and ABER positions (set 2). Two readers independently interpreted both sets, and diagnoses were compared with arthroscopic findings. Diagnostic accuracy was calculated, and 95% confidence intervals were used to detect significant differences between sets. Diagnostic confidence was recorded by using a three-level confidence score. Differences between sets were evaluated by using the Wilcoxon signed rank test. Interobserver agreement was determined separately for each set and for all diagnoses, full-thickness tears, and partial-thickness tears. RESULTS: For full thickness tears, there was no benefit to reading set 2. For reader 1, sensitivity and specificity were 95% and 100%, respectively, for set 1 and 100% and 100%, respectively, for set 2. For reader 2, sensitivity and specificity were 80% and 100%, respectively, for set 1 and 100% and 100%, respectively, for set 2. For partial-thickness tears, sensitivity was significantly higher after reading set 2. For reader 1, sensitivity and specificity were 71% and 88%, respectively, for set 1 and 93% and 100%, respectively, for set 2. For reader 2, sensitivity and specificity were 50% and 88%, respectively, for set 1 and 86% and 94%, respectively, for set 2. For both readers, diagnostic confidence for partial thickness tears was significantly higher after reading set 2. After the interpretation of set 2, kappa values increased from 0.35 to 1.00 for full thickness tears and from 0.12 to 0.63 for partial-thickness tears. CONCLUSION: Indirect MR arthrography with supplementary images obtained with patients in the ABER position significantly improved sensitivity and increased diagnostic confidence for partial-thickness tears of the supraspinatus tendon. Interobserver agreement was improved for both full- and partial-thickness tears. PMID- 16709791 TI - Aortic stenosis: comparative evaluation of 16-detector row CT and echocardiography. AB - PURPOSE: To prospectively evaluate whether planimetric measurements of aortic valve area (AVA) with 16-detector row computed tomography (CT) allow classification of aortic stenosis (AS). MATERIALS AND METHODS: The study had institutional review board approval; patients gave informed consent. Twenty patients (11 men, nine women; mean age, 63 years) with AS and 20 patients (10 men, 10 women; mean age, 65 years) without underwent transthoracic echocardiography (TTE), transesophageal echocardiography (TEE), and retrospectively electrocardiographically gated 16-detector row CT. Twenty CT data sets were reconstructed in 5% steps of R-R interval; data analysis was performed with four-dimensional software. Maximum AVA in systole planimetrically measured with CT (AVA(CT)) was compared with AVA planimetrically measured with TEE (AVA(TEE)), AVA calculated with the continuity equation and TTE (AVA(TTE)), and transvalvular pressure gradients determined with the Bernoulli equation and TTE. Correlations among AVA(CT), AVA(TTE), AVA(TEE), and transvalvular pressure gradients were tested with bivariate regression analysis; agreement between methods was assessed with the Bland-Altman method. RESULTS: In patients without AS, mean AVA(CT) was 3.56 cm2 +/- 0.66 and mean AVA(TEE) was 3.43 cm2 +/- 0.69. In patients with AS, mean AVA(CT) was 0.89 cm2 +/- 0.35; mean AVA(TEE), 0.86 cm2 +/- 0.35; and mean AVA(TTE), 0.83 cm2 +/- 0.33. Mean transvalvular pressure gradient was 51 mm Hg +/- 22. Significant correlations were present between AVA(CT) and AVA(TEE) (r = 0.99, P < .001), AVA(CT) and AVA(TTE) (r = 0.95, P < .001), and AVA(CT) and transvalvular pressure gradients (r = -0.74, P < .01). Mean differences were -0.08 cm2 (limits of agreement: -0.32, 0.16) for AVA(CT) versus AVA(TEE) and 0.06 cm2 (limits of agreement: -0.15, 0.26) for AVA(CT) versus AVA(TTE). CONCLUSION: Planimetric measurements of AVA with retrospectively electrocardiographically gated 16-detector row CT allow classification of AS that is similar to that achieved with measurements by using echocardiographic methods. PMID- 16709792 TI - Accuracy of stepping-table lower extremity MR angiography with dual-level bolus timing and separate calf acquisition: hybrid peripheral MR angiography. AB - PURPOSE: To retrospectively evaluate the accuracy of hybrid peripheral magnetic resonance (MR) angiography by using conventional digital subtraction angiography (DSA) as the reference standard. MATERIALS AND METHODS: This retrospective study protocol received approval from the Office of Sponsored Research at Northwestern University, which included review by the Office for the Protection of Research Subjects. Informed consent was waived for this HIPAA-compliant study. One hundred twenty-one consecutive patients (67 men: mean age, 66 years +/- 12 [standard deviation]; 54 women: mean age, 69 years +/- 14), who were referred for evaluation of peripheral vascular disease, underwent peripheral contrast material enhanced MR angiography. By using a hybrid technique, two independent timing measurements were performed in the pelvis and calves followed by MR angiography of the calves and, subsequently, a pelvis-thigh stepping-table acquisition. Images were evaluated for extent of disease, on the basis of degree of stenosis; for venous contamination, on the basis of venous signal intensity; and for diagnostic quality, on the basis of diagnostic confidence of the observer. DSA correlation of the extent of vascular disease was available in 45 of these patients, which was used to evaluate the diagnostic power of the hybrid technique. RESULTS: For detection of stenosis greater than 50%, the hybrid technique had 95% sensitivity (P < .05), 95% specificity (P < .05), and 95% accuracy (P < .05). There was no significant venous contamination in any of the examinations performed with this technique. CONCLUSION: The hybrid peripheral MR angiography technique provides diagnostic-quality examinations and virtually eliminates venous contamination. PMID- 16709793 TI - Renal masses in the adult patient: the role of percutaneous biopsy. AB - Although percutaneous renal mass biopsy with cross-sectional imaging guidance has long been considered to be safe and accurate, there have been recent advances in imaging, interventional, and cytologic techniques that have increased the role of percutaneous biopsy in the diagnosis of renal masses. Today, biopsy plays a fundamental role in the care of patients with a renal mass. Biopsy results are used to confirm the diagnosis of renal cancers, metastases, and infections, and there is increasing evidence to suggest that biopsy can help subtype and grade many primary renal cancers. Because a considerable fraction of small solid renal masses are benign and do not need treatment, there is an increasing need to diagnose them. Biopsy after a full imaging work-up can help prevent unnecessary and potentially morbid surgical and ablation procedures in a substantial number of patients. Although more data are needed to understand the overall accuracy of biopsy for the diagnosis of benign lesions, many can be diagnosed with the aid of biopsy findings. This article reviews reported experience with percutaneous renal mass biopsy, discusses the technical factors that contribute to results, and details seven specific clinical settings that should prompt the clinician to consider percutaneous biopsy when encountering a renal mass. PMID- 16709794 TI - The divergence of lymphocyte development pathways. PMID- 16709795 TI - The functions of the thymus system and the bursa system in the chicken. 1966. PMID- 16709796 TI - Space, selection, and surveillance: setting boundaries with BLyS. AB - The BLyS family of ligands and receptors governs B cell homeostasis by controlling survival, differentiation, and lifespan. This family consists of multiple receptors and ligands, allowing independent regulation of different B cell subsets by varying the combination and levels of receptors expressed. Multiple downstream signaling pathways are implicated in these activities, reflecting this receptor complexity as well as cross-talk with other B cell signaling systems. BLyS levels are associated with multiple forms of humoral autoimmunity and can modulate tolerogenic elimination at the transitional checkpoint. BLyS responsiveness thus balances peripheral selection against cell numbers, providing an elastic system that varies selective stringency based on homeostatic demands. PMID- 16709797 TI - Cutting edge: T cell Ig mucin-3 reduces inflammatory heart disease by increasing CTLA-4 during innate immunity. AB - Autoimmune diseases can be reduced or even prevented if proinflammatory immune responses are appropriately down-regulated. Receptors (such as CTLA-4), cytokines (such as TGF-beta), and specialized cells (such as CD4+CD25+ T regulatory cells) work together to keep immune responses in check. T cell Ig mucin (Tim) family proteins are key regulators of inflammation, providing an inhibitory signal that dampens proinflammatory responses and thereby reducing autoimmune and allergic responses. We show in this study that reducing Tim-3 signaling during the innate immune response to viral infection in BALB/c mice reduces CD80 costimulatory molecule expression on mast cells and macrophages and reduces innate CTLA-4 levels in CD4+ T cells, resulting in decreased T regulatory cell populations and increased inflammatory heart disease. These results indicate that regulation of inflammation in the heart begins during innate immunity and that Tim-3 signaling on cells of the innate immune system critically influences regulation of the adaptive immune response. PMID- 16709798 TI - Translocation of active heparanase to cell surface regulates degradation of extracellular matrix heparan sulfate upon transmigration of mature monocyte derived dendritic cells. AB - After Ag capture and exposure to danger stimuli, maturing dendritic cells (DCs) migrate to regional lymph nodes, where antigenic peptides are presented to T lymphocytes. To migrate from peripheral tissue such as the epidermis to regional lymph nodes, Ag-bearing epidermal Langerhans cells must move through an extracellular matrix (ECM) of various compositions. The nature of their capacity to transmigrate via ECM is not well understood, although MIP-3beta and CCR7 play critical roles. We were interested in verifying whether heparanase, a heparan sulfate-degrading endo-beta-d-glucuronidase that participates in ECM degradation and remodeling, is expressed and functional in monocyte-derived DCs. Using immunohistochemistry, confocal microscopy, RT-PCR, Western blot analysis, assays for heparanase activity, and Matrigel transmigration, we show that heparanase is expressed in both nuclei and cytoplasm of immature DCs, and that gene expression and synthesis take place mainly in monocytes and early immature DCs. We also found that both nuclear and cytoplasm fractions show heparanase activity, and upon LPS-induced maturation, heparanase translocates to the cell surface and degrades ECM heparan sulfate. Matrigel transmigration assays showed a MIP-3beta comparable role for heparanase. Because heparan sulfate glycosaminoglycans play a key role in the self-assembly, insolubility, and barrier properties of the ECM, the results of this study suggest that heparanase is a key enzyme in DC transmigration through the ECM. PMID- 16709799 TI - Monocyte/macrophage dysfunctions do not impair the promotion of myelofibrosis by high levels of thrombopoietin. AB - Several lines of evidence indicate that the megakaryocyte/platelet lineage is crucial in myelofibrosis induction. The demonstration that NOD/SCID mice with functionally deficient monocytes do not develop fibrotic changes when exposed to thrombopoietin (TPO) also suggests an important role for monocyte/macrophages. However, in this animal model, the development of myelofibrosis is dependent on the level of TPO. This study was conducted to investigate whether NOD/SCID mice exposed to high TPO levels mediated by a retroviral vector would be refractory to the development of bone marrow fibrosis. We show that TPO and TGF-beta1 in plasma from NOD/SCID and SCID mice engrafted with TPO-overexpressing hemopoietic cells reach levels similar to the ones reached in immunocompetent mice, and all animals develop a myeloproliferative disease associated with a dense myelofibrosis at 8 wk posttransplantation. Monocytes in NOD/SCID mice are functionally deficient to secrete cytokines such as IL-1alpha in response to stimuli, even under TPO expression. Surprisingly, the plasma of these mice displays high levels of IL alpha, which was demonstrated to originate from platelets. Together, these data suggest that completely functional monocytes are not required to develop myelofibrosis and that platelets are able, under TPO stimulation, to synthesize inflammatory cytokines, which may be involved in the pathogenesis of myelofibrosis and osteosclerosis. PMID- 16709800 TI - Glucocorticoid-induced TNF receptor family related gene activation overcomes tolerance/ignorance to melanoma differentiation antigens and enhances antitumor immunity. AB - Glucocorticoid-induced TNF receptor family related protein (GITR) is present on many different cell types. Previous studies have shown that in vivo administration of an anti-GITR agonist mAb (DTA-1) inhibits regulatory T cells (Treg)-dependent suppression and enhances T cell responses. In this study, we show that administration of DTA-1 induces >85% tumor rejection in mice challenged with B16 melanoma. Rejection requires CD4+, CD8+, and NK1.1+ cells and is dependent on IFN-gamma and Fas ligand and independent of perforin. Depletion of Treg via anti-CD25 treatment does not induce B16 rejection, whereas 100% of the mice depleted of CD25+ cells and treated with DTA-1 reject tumors, indicating a predominant role of GITR on effector T cell costimulation rather than on Treg modulation. T cells isolated from DTA-1-treated mice challenged with B16 are specific against B16 and several melanoma differentiation Ags. These mice develop memory against B16, and a small proportion of them develop mild hypopigmentation. Consistent with previous studies showing that GITR stimulation increases Treg proliferation in vitro, we found in our model that GITR stimulation expanded the absolute number of FoxP3+ cells in vivo. Thus, we conclude that overall, GITR stimulation overcomes self-tolerance/ignorance and enhances T cell-mediated antitumor activity with minimal autoimmunity. PMID- 16709801 TI - Suppression of alveolar macrophage apoptosis prolongs survival of rats and mice with pneumocystis pneumonia. AB - The number of alveolar macrophages is decreased in patients or animals with Pneumocystis pneumonia (Pcp). This loss of alveolar macrophages is in part due to apoptosis caused by Pneumocystis infection. The mechanism of apoptosis induction is unknown. Cell-free bronchoalveolar lavage fluids from Pneumocystis-infected rats or mice have the ability to induce apoptosis in normal alveolar macrophages. To characterize the mechanisms by which apoptosis proceeds in alveolar macrophages during Pcp, specific caspase inhibitors are tested for their ability to suppress the apoptosis. In vitro induction of apoptosis can be inhibited by the caspase-9 inhibitor (Z-LEHD-FMK) but not by the inhibitor to caspase-8 or 10. The caspase-9 inhibitor can also inhibit apoptosis of alveolar macrophages in vivo when it is intranasally instilled into dexamethasone-immunosuppressed, Pneumocystis-infected rats or L3T4 cell-depleted, Pneumocystis-infected mice. The number of alveolar macrophages rebounds in caspase-9 inhibitor-treated Pcp animals. Phagocytic activity of alveolar macrophages in treated animals is also recovered, and organism burden in these animals is reduced. Administration of caspase-9 inhibitor also clears the exudate that normally fills the alveoli during Pcp and decreases lung inflammation. Furthermore, caspase-9-treated Pcp animals survive for the entire 70-day period of the study, whereas nontreated Pcp animals die 40-60 days after initiation of infection. Depletion of recovered alveolar macrophages by intranasal administration of clodronate-containing liposomes in caspase-9 inhibitor-treated animals abrogates the effects of the inhibitor. Together, these results indicate that immunomodulation of the host response may be an alternative to current treatments for Pcp. PMID- 16709802 TI - NK lytic-associated molecule, involved in NK cytotoxic function, is an E3 ligase. AB - NK lytic-associated molecule (NKLAM) is a protein involved in the cytolytic function of NK cells and CTLs. It has been localized to the cytolytic granules in NK cells and is up-regulated when cells are exposed to cytokines IL-2 or IFN beta. We report in this study that NKLAM contains a cysteine-rich really interesting new gene (RING) in between RING-RING domain, and that this domain possesses strong homology to the RING domain of the known E3 ubiquitin ligase, Dorfin. To determine whether NKLAM functions as an E3 ligase, we performed coimmunoprecipitation binding assays with ubiquitin conjugates (Ubcs) UbcH7, UbcH8, and UbcH10. We demonstrated that both UbcH7 and UbcH8 bind to full-length NKLAM. We then performed a similar binding assay using endogenous NKLAM and UbcH8 expressed by human NK clone NK3.3 to show that the protein interaction occurs in vivo. Using the yeast two-hybrid system, we identified uridine kinase like-1 (URKL-1) protein as a substrate for NKLAM. We confirmed that NKLAM and URKL-1 interact in mammalian cells by using both immunoprecipitation and confocal microscopy. We demonstrated decreased protein expression and enhanced ubiquitination of URKL-1 in the presence of NKLAM. These data indicate that NKLAM is a RING finger protein that binds Ubcs and has as one of its substrates, URKL 1, thus defining this cytolytic protein as an E3 ubiquitin ligase. PMID- 16709803 TI - Selective export of HLA-F by its cytoplasmic tail. AB - MHC class I molecules exit the endoplasmic reticulum (ER) by an unknown mechanism. Although a selective export mechanism has been proposed for the anterograde transport of class I, a motif responsible for export has never been identified. Although classical class I molecules lacking their cytoplasmic tail are expressed on the cell surface, we found that HLA-F was entirely dependent on its cytoplasmic tail for export from the ER. Two known export motifs were recognizable in HLA-F. A C-terminal valine residue functioned in ER export and interacted with coat complex (COP)II, while an RxR motif also played an important role in anterograde transport and bound to 14-3-3 proteins. This divergent trafficking of HLA-F implicates an alternative function for HLA-F, independent of loading with peptides in the ER. PMID- 16709804 TI - Functional and molecular comparison of anergic and regulatory T lymphocytes. AB - Tolerance in vivo is maintained by multiple mechanisms that function to prevent autoimmunity. An encounter of CD4+ T cells with a circulating self-Ag leads to partial thymic deletion, the development of CD25+ regulatory T cells (Tregs), and functional anergy in the surviving CD25- population. We have compared anergic and regulatory T cells of the same Ag specificity generated in vivo by the systemic self-Ag. Anergic cells are unresponsive to the self-Ag that induces tolerance, but upon transfer into a new host and immunization, anergic cells can induce a pathologic autoimmune reaction against tissue expressing the same Ag. Tregs, in contrast, are incapable of mediating harmful reactions. To define the basis of this functional difference, we have compared gene expression profiles of anergic and regulatory T cells. These analyses show that Tregs express a distinct molecular signature, but anergic cells largely lack such a profile. Anergic cells express transcripts that are associated with effector differentiation, e.g., the effector cytokines IL-4 and IFN-gamma. Anergic cells do not produce these cytokines in response to self-Ag, because the cells exhibit a proximal signaling block in response to TCR engagement. Thus, anergy reflects an aborted activation pathway that can readily be reversed, resulting in pathologic effector cell responses, whereas Treg development follows a distinct developmental pathway that extinguishes effector functions. PMID- 16709805 TI - Cervical thymus in the mouse. AB - Although thymic ectopy has long been recognized in humans, the functional activity or potential immunological significance of this thymic tissue is unknown. In this study, we describe murine thymic ectopy, cervical thymic tissue that possesses the same general organization as the thoracic thymus, that is able to support T cell differentiation, and that can export T cells to the periphery. Unexpectedly, the pattern of autoantigen expression by ectopic thymic tissue differs from that of the thoracic thymus, raising the possibility that these two thymic environments may project different versions of self. PMID- 16709806 TI - Exogenous and endogenous TLR ligands activate anti-chromatin and polyreactive B cells. AB - Autoreactive B cells may become activated in a T-independent manner via synergistic engagement of the BCR and TLRs. Using the VH3H9 Ig H chain transgene to track anti-chromatin B cells, we demonstrate that VH3H9/Vlambda1 anti chromatin B cells proliferate in response to stimulatory oligodeoxynucleotides containing CpG motifs, suggesting that these autoreactive B cells are responsive to TLR9 signaling. Strikingly, some VH3H9 B cells, but not the well-characterized VH3H9/Vlambda1 B cells, proliferate spontaneously in culture medium. This proliferation is blocked by inhibitory CpG oligodeoxynucleotides, implicating the TLR9 (or possibly TLR7) pathway. Most hybridomas generated from the proliferating cells are polyreactive, and one exhibits binding to nuclear Ags but not to the other Ags tested. Thus, B cells carrying autoreactive and/or polyreactive specificities may be susceptible to T cell-independent activation via dual engagement of the BCR and TLRs. PMID- 16709807 TI - Conditional ablation of MHC-II suggests an indirect role for MHC-II in regulatory CD4 T cell maintenance. AB - Although the importance of MHC class II (MHC-II) in acute homeostatic proliferation of regulatory T (Treg) cells has been established, we considered here the maintenance and state of Treg cells in mice that are almost completely devoid of MHC-II in their periphery but still make their own CD4 T cells and Treg cells. The latter was accomplished by conditional deletion of a loxP-flanked MHC II beta-chain allele using a TIE2Cre transgene, which causes a very high degree of deletion in hemopoietic/endothelial progenitor cells but without deletion among thymic epithelial cells. Such conditional MHC-II-deficient mice possess their own relatively stable levels of CD4+CD25+ cells, with a normal fraction of Foxp3+ Treg cells therein, but at a level approximately 2-fold lower than in control mice. Thus, both Foxp3low/- CD4+CD25+ cells, said to be a major source of IL-2, and IL-2-dependent Foxp3+ Treg cells are reduced in number. Furthermore, CD25 expression is marginally reduced among Foxp3+ Treg cells in conditional MHC II-deficient mice, indicative of a lack of MHC-II-dependent TCR stimulation and/or IL-2 availability, and IL-2 administration in vivo caused greatly increased cell division among adoptively transferred Treg cells. This is not to say that IL-2 can cause Treg cell division in the complete absence of MHC-II as small numbers of MHC-II-bearing cells do remain in conditional MHC-II-deficient mice. Rather, this suggests only that IL-2 was limiting. Thus, our findings lend support to the proposal that Treg cell homeostasis depends on a delicate balance with a population of self-reactive IL-2-producing CD4+CD25+ cells which are themselves at least in part MHC-II-dependent. PMID- 16709809 TI - Splenic atrophy in experimental stroke is accompanied by increased regulatory T cells and circulating macrophages. AB - Induction of stroke not only produces local ischemia and brain damage, but also has profound effects on peripheral immune responses. In the current study, we evaluated effects on spleen and blood cells 4 days after stroke induction. Surprisingly, there was a less inflammatory cytokine profile in the middle cerebral artery occlusion-affected right brain hemisphere at 96 h compared with earlier time points. Moreover, our results demonstrate that stroke leads to splenic atrophy characterized by a reduction in organ size, a drastic loss of splenocyte numbers, and induction of annexin V+ and TUNEL+ cells within the spleen that are in the late stages of apoptosis. The consequence of this process was to reduce T cell proliferation responses and secretion of inflammatory cytokines, resulting in a state of profound immunosuppression. These changes produced a drastic reduction in B cell numbers in spleen and blood, and a novel increase in CD4+FoxP3+ regulatory T cells. Moreover, we detected a striking increase in the percentage of nonapoptotic CD11b+ VLA-4-negative macrophages/monocytes in blood. Immunosuppression in response to brain injury may account for the reduction of inflammatory factors in the stroke-affected brain, but also potentially could curtail protective immune responses in the periphery. These findings provide new evidence to support the contention that damage to the brain caused by cerebral ischemia provides a powerful negative signal to the peripheral immune system that ultimately induces a drastic state of immunosuppression caused by cell death as well as an increased presence of CD4+FoxP3+ regulatory T cells. PMID- 16709808 TI - Function of liver activation-regulated chemokine/CC chemokine ligand 20 is differently affected by cathepsin B and cathepsin D processing. AB - Chemokine processing by proteases is emerging as an important regulatory mechanism of leukocyte functions and possibly also of cancer progression. We screened a large panel of chemokines for degradation by cathepsins B and D, two proteases involved in tumor progression. Among the few substrates processed by both proteases, we focused on CCL20, the unique chemokine ligand of CCR6 that is expressed on immature dendritic cells and subtypes of memory lymphocytes. Analysis of the cleavage sites demonstrate that cathepsin B specifically cleaves off four C-terminally located amino acids and generates a CCL20(1-66) isoform with full functional activity. By contrast, cathepsin D totally inactivates the chemotactic potency of CCL20 by generating CCL20(1-55), CCL20(1-52), and a 12-aa C-terminal peptide CCL20(59-70). Proteolytic cleavage of CCL20 occurs also with chemokine bound to glycosaminoglycans. In addition, we characterized human melanoma cells as a novel CCL20 source and as cathepsin producers. CCL20 production was up-regulated by IL-1alpha and TNF-alpha in all cell lines tested, and in human metastatic melanoma cells. Whereas cathepsin D is secreted in the extracellular milieu, cathepsin B activity is confined to cytosol and cellular membranes. Our studies suggest that CCL20 processing in the extracellular environment of melanoma cells is exclusively mediated by cathepsin D. Thus, we propose a model where cathepsin D inactivates CCL20 and possibly prevents the establishment of an effective antitumoral immune response in melanomas. PMID- 16709810 TI - T cells in cryptopatch aggregates share TCR gamma variable region junctional sequences with gamma delta T cells in the small intestinal epithelium of mice. AB - The role of cryptopatch aggregates in the development of intestinal intraepithelial lymphocytes (IEL) is a matter of controversy. Therefore, an important question is whether T cells in cryptopatch aggregates are lineally related to IEL. We hypothesized that if gammadelta+ IEL derive from T cells in cryptopatch aggregates, then a clonal relationship would exist between the two populations. To test this hypothesis, we compared the sequence of rearranged TCR gamma variable region 5 genes in gammadelta+ IEL and cryptopatch cells. We purified IEL by FACS and cryptopatch cells were isolated from frozen sections of the intestine by laser-assisted microdissection. PCR showed that TCR gamma variable region 5 was rearranged in gammadelta+ IEL and in CD3+ cryptopatch cells, but not in CD3- cryptopatch cells. DNA sequence analysis showed that the frequency of in-frame junctions in cryptopatch aggregates was at a level consistent with positive selection in both wild-type and athymic nude mice. In addition, the predicted amino acid sequences of V-J junctions present in gammadelta+ IEL and cryptopatch cells were encoded by identical nucleotide sequences. By contrast, the frequency of in-frame joints was significantly reduced in cryptopatch cells isolated from TCR delta-deficient mice, indicating that the enrichment of in-frame joints in cryptopatch cells must normally depend on expression of surface gammadelta TCR. Our results are consistent with the hypothesis that a subset of gammadelta+ IEL are related to T cells in cryptopatch aggregates. The precise role of cryptopatch aggregates in intestinal gammadelta+ T cell homeostasis still needs to be determined. PMID- 16709811 TI - Stimulation through CD40 on mouse and human renal cell carcinomas triggers cytokine production, leukocyte recruitment, and antitumor responses that can be independent of host CD40 expression. AB - CD40, a member of the TNFR superfamily, is expressed on a variety of host immune cells, as well as some tumors. In this study, we show that stimulation of CD40 expressed on both mouse and human renal carcinoma cells (RCCs) triggers biological effects in vitro and in vivo. Treatment of the CD40+ Renca mouse RCC tumor cells in vitro with an agonistic anti-CD40 Ab induced strong expression of the genes and proteins for GM-CSF and MCP-1, and induced potent chemotactic activity. Similarly, administration of alphaCD40 to both wild-type and CD40-/- mice bearing Renca tumors resulted in substantial amounts of TNF-alpha and MCP-1 in the serum, increased the number of total splenocytes and MHC class II+ CD11c+ leukocytes, and when combined with IFN-gamma, inhibited the progression of established Renca tumors in vivo in both wild-type and CD40-/- mice. Similarly, treatment of CD40+ A704 and ACHN human RCC lines with mouse anti-human CD40 Ab induced strong expression of genes and proteins for MCP-1, IL-8, and GM-CSF in vitro and in vivo. Finally, in SCID mice, the numbers of ACHN pulmonary metastases were dramatically reduced by treatment with species-specific human CD40 Ab. These results show that CD40 stimulation of CD40+ tumor cells can enhance immune responses and result in antitumor activity. PMID- 16709812 TI - TCR gamma delta intraepithelial lymphocytes are required for self-tolerance. AB - Neonatal thymectomy (NTX) impairs T cell regulation and leads to organ-specific autoimmune disease in susceptible mouse strains. In the NOD mouse model of spontaneous type 1 diabetes, we observed that NTX dramatically accelerated autoimmune pancreatic beta cell destruction and diabetes. NTX had only a minor effect in NOD mice protected from diabetes by transgenic expression of the beta cell autoantigen proinsulin in APCs, inferring that accelerated diabetes after NTX is largely due to failure to regulate proinsulin-specific T cells. NTX markedly impaired the development of intraepithelial lymphocytes (IEL), the number of which was already reduced in euthymic NOD mice compared with control strains. IEL purified from euthymic NOD mice, specifically CD8alphaalpha TCRgammadelta IEL, when transferred into NTX-NOD mice, trafficked to the small intestinal epithelium and prevented diabetes. Transfer of prototypic CD4+CD25+ regulatory T cells also prevented diabetes in NTX-NOD mice; however, the induction of these cells by oral insulin in euthymic mice depended on the integrity of TCRgammadelta IEL. We conclude that TCRgammadelta IEL at the mucosal interface between self and nonself play a key role in maintaining peripheral tolerance both physiologically and during oral tolerance induction. PMID- 16709813 TI - Impact of orthologous melan-A peptide immunizations on the anti-self melan-A/HLA A2 T cell cross-reactivity. AB - In HLA-A2 individuals, the CD8 T cell response against the differentiation Ag Melan-A is mainly directed toward the peptide Melan-A26-35. The murine Melan-A24 33 sequence encodes a peptide that is identical with the human Melan-A26-35 decamer, except for a Thr-to-Ile substitution at the penultimate position. Here, we show that the murine Melan-A24-33 is naturally processed and presented by HLA A2 molecules. Based on these findings, we compared the CD8 T cell response to human and murine Melan-A peptide by immunizing HLA-A2 transgenic mice. Even though the magnitude of the CTL response elicited by the murine Melan-A peptide was lower than the one elicited by the human Melan-A peptide, both populations of CTL recognized the corresponding immunizing peptide with the same functional avidity. Interestingly, CTL specific for the murine Melan-A peptide were completely cross-reactive against the orthologous human peptide, whereas anti human Melan-A CTL recognized the murine Melan-A peptide with lower avidity. Structurally, this discrepancy could be explained by the fact that Ile32 of murine Melan-A24-33 created a larger TCR contact area than Thr34 of human Melan A26-35. These data indicate that, even if immunizations with orthologous peptides can induce strong specific T cell responses, the quality of this response against syngeneic targets might be suboptimal due to the structure of the peptide-TCR contact surface. PMID- 16709814 TI - Abrogation of functional selectin-ligand expression reduces migration of pathogenic CD8+ T cells into heart. AB - CD8+ T cells are involved in autoimmune and infectious myocarditis and cardiac allograft rejection. The role of selectins in cardiac recruitment of CD8+ T cells is not understood. In this study, the contribution of T cell selectin ligands to effector CD8+ T cell recruitment into the heart was examined using a model of myocarditis, which depends on transfer of OVA peptide-specific CD8+ T cells (OT I) into mice (CMy-mOva) that express OVA in the heart. alpha-(1,3) Fucosyltransferase (FucT)-VII-deficient OT-I cells displayed over a 95% reduction in their ability to interact with P-selectin under flow conditions in vitro, compared with wild-type OT-I cells. Interaction of FucT-VII-deficient OT-I cells with E-selectin was reduced approximately 50%. FucT-VII-deficient OT-I cells were also less efficiently recruited into a dermal site of Ag and adjuvant injection. Significantly, FucT-VII-deficient OT-I cells were also impaired in their ability to migrate into CMy-mOva hearts, compared with wild-type OT-I cells. Transfer of FucT-VII-deficient T cells caused less severe early myocarditis and myocyte damage than transfer of wild-type T cells. Combined FucT-IV/VII-deficient OT-I cells displayed a more profound reduction in E-selectin interactions in vitro compared with FucT-VII-deficient T cells, and the FucT-IV/VII-deficient T cells also showed less early recruitment and pathogenicity in the CMy-mOva myocarditis model. These results identify a prominent role for selectin ligands in contributing to effector CD8+ T cell recruitment into the myocardium and indicate that selectin-dependent T cell recruitment is relevant to other tissues besides the skin. PMID- 16709815 TI - A complex of Wiskott-Aldrich syndrome protein with mammalian verprolins plays an important role in monocyte chemotaxis. AB - The Wiskott-Aldrich syndrome protein (WASP) is a product of the gene defective in an Xid disorder, Wiskott-Aldrich syndrome. WASP expression is limited to hemopoietic cells, and WASP regulates the actin cytoskeleton. It has been reported that monocytes/macrophages from WASP-deficient Wiskott-Aldrich syndrome patients are severely defective in chemotaxis, resulting in recurrent infection. However, the molecular basis of such chemotactic defects is not understood. Recently, the WASP N-terminal region was found to bind to the three mammalian verprolin homologs: WASP interacting protein (WIP); WIP and CR16 homologous protein (WICH)/WIP-related protein (WIRE); and CR16. Verprolin was originally found to play an important role in the regulation of actin cytoskeleton in yeast. We have shown that WASP, WIP, and WICH/WIRE are expressed predominantly in the human monocyte cell line THP-1 and that WIP and WICH/WIRE are involved in monocyte chemotaxis. When WASP binding to verprolins was blocked, chemotactic migration of monocytes was impaired in both THP-1 cells and primary human monocytes. Increased expression of WASP and WIP enhanced monocyte chemotaxis. Blocking WASP binding to verprolins impaired cell polarization but not actin polymerization. These results indicate that a complex of WASP with mammalian verprolins plays an important role in chemotaxis of monocytes. Our results suggest that WASP and mammalian verprolins function as a unit in monocyte chemotaxis and that the activity of this unit is critical to establish cell polarization. In addition, our results also indicate that the WASP-verprolin complex is involved in other functions such as podosome formation and phagocytosis. PMID- 16709816 TI - CD4+CD25+Foxp3+ T cells and CD4+CD25-Foxp3+ T cells in aged mice. AB - Aging is associated with a progressive decline in T cell-mediated immune responses. However, it has been unknown whether regulatory/suppressive CD4 T cells are involved in this decline. Our in vitro analyses revealed that CD4+CD25+ T cells, the well-characterized naturally occurring regulatory/suppressive CD4 T cells, in aged mice are functionally comparable to those in young mice (i.e., anergic and suppressive), although slightly increased in number. In contrast, functional changes to whole CD4+CD25- T cells were pronounced in aged mice, i.e., the majority of aged CD4+CD25- T cells exhibited a significant hyporesponsiveness, and the remaining cells maintained a normal responsiveness. Furthermore, we identified Foxp3 (a transcription factor critical in conferring the regulatory/suppressive function to CD4 T cells)-positive suppressive CD4 T cells among aged hyporesponsive CD4+CD25- T cells. These results suggest that the age-related decline in T cell-mediated immune responses is ascribable to changes in the CD4+CD25- T cell population and not to a functional augmentation of suppressive CD4+CD25+ T cells. PMID- 16709817 TI - Macrophage-colony-stimulating factor-induced proliferation and lipopolysaccharide dependent activation of macrophages requires Raf-1 phosphorylation to induce mitogen kinase phosphatase-1 expression. AB - Macrophages are key regulators of immune responses. In the absence of an activating signal, murine bone marrow-derived macrophages undergo proliferation in response to their specific growth factor, namely M-CSF. The addition of bacterial LPS results in macrophage growth arrest and their engagement in a proinflammatory response. Although participation of ERKs is required for both macrophage proliferation and activation, ERK phosphorylation follows a more delayed pattern in response to activating agents. In primary macrophages, mitogen kinase phosphatase-1 (MKP-1) is a key regulator of the time course of MAPK activity. Here we showed that MKP-1 expression is dependent on Raf-1 activation. The time course of Raf-1 activation correlated with that of ERK-1/2. However, whereas ERK phosphorylation in response to M-CSF is Raf-1 dependent, in response to LPS, an alternative pathway directs the activation of these kinases. Inhibition of Raf-1 activity increased the expression of cyclin-dependent kinase inhibitors and growth arrest. In contrast, no effect was observed in the expression of proinflammatory cytokines and inducible NO synthase following LPS stimulation. The data reported here reveal new insights into how signaling determines opposing macrophage functions. PMID- 16709818 TI - B and T lymphocyte attenuator-mediated signal transduction provides a potent inhibitory signal to primary human CD4 T cells that can be initiated by multiple phosphotyrosine motifs. AB - The B and T lymphocyte attenuator (BTLA) is a recently identified member of the CD28 family of cell receptors. Initial reports demonstrated that mice deficient in BTLA expression were more susceptible to experimental autoimmune encephalomyelitis, indicating that BTLA was likely to function as a negative regulator of T cell activation. However, cross-linking of BTLA only resulted in a 2-fold reduction of IL-2 production, questioning the potency with which BTLA engagement blocks T cell activation. We established a model in which BTLA signaling could be studied in primary human CD4 T cells. We observed that cross linking of a chimeric receptor consisting of the murine CD28 extracellular domain and human BTLA cytoplasmic tail potently inhibits IL-2 production and completely suppresses T cell expansion. Mutation of any BTLA tyrosine motifs had no effect on the ability of BTLA to block T cell activation. Only mutation of all four tyrosines rendered the BTLA cytoplasmic tail nonfunctional. We performed structure-function studies to determine which factors recruited to the BTLA cytoplasmic tail correlated with BTLA function. Using pervanadate as a means to phosphorylate the BTLA cytoplasmic tail, we observed both Src homology protein (SHP)-1 and SHP-2 recruitment. However, upon receptor engagement, we observed only SHP-1 recruitment, and mutations that abrogated SHP-1 recruitment did not impair BTLA function. These studies question whether SHP-1 or SHP-2 have any role in BTLA function and caution against the use of pervanadate as means to initiate signal transduction cascades in primary cells. PMID- 16709819 TI - Regulation of Ly49D/DAP12 signal transduction by Src-family kinases and CD45. AB - Activating, DAP12-coupled members of the Ly-49 family of NK cell receptors help control viral infections in mice. However, the kinases and/or phosphatases mediating tyrosine phosphorylation of Ly-49D-associated DAP12 have not been elucidated. In this study, we show for the first time that Src family tyrosine kinases are physically and functionally associated with Ly-49D/DAP12 signaling in murine NK cells. Specifically, we demonstrate the following: 1) inhibition of Src family kinases suppresses DAP12 phosphorylation and downstream DAP12 signals; 2) both Fyn and Lck are capable of phosphorylating DAP12; and 3) both kinases coimmunoprecipitate with the Ly-49D/DAP12 complex in NK cells. Although we detect enhanced phosphorylation of Fyn upon Ly-49D cross-linking in NK cells, Ly-49D mediated events in both Fyn-/- and Fyn/Lck-/- mice appear normal, reinforcing the theme of redundancy in the ability of Src family kinases to initiate activation events. In contrast to disruption of specific Src family enzymes, Ly-49D/DAP12 mediated calcium mobilization and cytokine production by CD45 null NK cells are defective. Although others have ascribed the effects of CD45 mutation solely on the suppression of Src family activity, we demonstrate in this study that DAP12 is hyperphosphorylated in CD45 null NK cells, resulting in uncoordinated tyrosine mediated signaling upon Ly-49D ligation. Therefore, although our data are consistent with a Src kinase activity proximally within DAP12 signaling, DAP12 also appears to be a substrate of CD45, suggesting a more complex role for this phosphatase than has been reported previously. PMID- 16709820 TI - Antitumor activity of the TLR-5 ligand flagellin in mouse models of cancer. AB - Flagellin, the structural protein subunit of the bacterial flagellum, is specifically recognized by TLR-5 and has potent immunomodulatory effects. The antitumor effects of purified Salmonella typhimurium flagellin were evaluated in mice transplanted s.c. with a weakly immunogenic murine tumor or with its variant stably transfected to express the highly antigenic human HER-2 oncoprotein. Peritumoral administration of flagellin 8-10 days after tumor implantation did not affect the growth rate of the weakly immunogenic tumor but significantly inhibited growth of the antigenic variant tumor. In contrast, flagellin administered at the time of implantation of the antigenic tumor led to accelerated tumor growth. These contrasting effects of flagellin on tumor growth correlated with the type of immune response induced; i.e., late flagellin administration was associated with an increased IFN-gamma:IL-4 ratio and the decreased frequency of CD4+CD25+ T regulatory cells, whereas flagellin treatment at the time of tumor implantation decreased the IFN-gamma:IL-4 ratio and increased CD4+CD25+ T cell frequency. When the early flagellin treatment was combined with administration of CpG-containing oligodeoxynucleotides, tumor growth was completely suppressed, indicating synergy between flagellin and CpG containing oligodeoxynucleotides. Together, these data provide evidence that flagellin can have contrasting effects on tumor growth. PMID- 16709821 TI - Functional regulatory T cells are collected in stem cell autografts by mobilization with high-dose cyclophosphamide and granulocyte colony-stimulating factor. AB - High-dose cyclophosphamide (Cy) and G-CSF are widely used to mobilize hemopoietic stem cells for treating patients with high-dose chemotherapy and autologous stem cell transplantation (ASCT). Because lymphocyte count in the graft collected after Cy-G-CSF treatment is an independent survival factor after ASCT for patients with multiple myeloma, our purpose was to study how Cy-G-CSF treatment affects the phenotype and function of T cells in patients with multiple myeloma. Cy induced a 3-fold decrease of T cell counts with a slow and partial T cell recovery of one-third at the time of hemopoietic stem cell collection. Cy-G-CSF treatment did not affect the relative ratios of central memory, effector memory, and late effector CD4+ or CD8+ T cells, but a decrease in the percentage of naive CD4+ cells was observed. The percentages of CD25+ cells increased 2- to 3-fold in CD4+ and CD8+ T cells, the former including both activated CD25low and CD25high cells. CD4+CD25high cells were regulatory T cells (Treg) that expressed high levels of FOXP3, CTLA-4, and GITR and displayed in vitro suppressive properties. The recovery of Treg absolute counts after Cy-G-CSF treatment was higher than the recovery of other lymphocyte subpopulations. In conclusion, Cy-G-CSF treatment induces a severe T cell count decrease without deleting Treg, which are potent inhibitors of antitumor response. The present data encourage novel therapeutic strategies to improve T cell recovery following ASCT while limiting Treg expansion. PMID- 16709822 TI - Differential signaling of T cell generation of IL-4 by wild-type and short deletion variant of type 2 G protein-coupled receptor for vasoactive intestinal peptide (VPAC2). AB - Vasoactive intestinal peptide (VIP) released from some neurons and T cells affects T cell migration, cytokine generation, and other functions by binding to constitutively expressed type 1 G protein-coupled receptor (VPAC1) or activation induced type 2 G protein-coupled receptor (VPAC2). Recently, a short-deletion (SD) splice variant of mouse VPAC2 that lacks 14 amino acids at the end of the last transmembrane domain has been identified in T cells and shown to resemble wild-type (WT) VPAC2 in affinity of VIP binding but to differ by lack of signaling of T cell adenylyl cyclase, migration, and IL-2 secretion. As Th2 cells are the principal source of immune VIP and have the greatest functional responses to VIP, the differences in signals transduced by WT and SD VPAC2 were studied in VPAC2-low D10G4.1 model Th2 cell transfectants individually expressing the respective types of VPAC2 equally. WT and SD VPAC2 Th2 cell transfectants secreted equal amounts of VIP. WT VPAC2 transfectants generated more IL-4 than did SD VPAC2 transfectants, and this increment was dependent on endogenous VIP. Exogenous VIP further increased IL-4 production by WT VPAC2 transfectants but decreased IL-4 production by SD VPAC2 transfectants. Cotransfection of the two constructs diminished VIP enhancement of IL-4 production seen with WT VPAC2 alone by preventing increases in nuclear levels of the requisite transcription factors c-Maf and Jun B. Thus the ratio of two forms of T cell VPAC2 determines the net effect of VIP on IL-4 generation by activated Th2 cells. PMID- 16709823 TI - Lipopolysaccharide-induced IL-1 beta production by human uterine macrophages up regulates uterine epithelial cell expression of human beta-defensin 2. AB - The uterine endometrium coordinates a wide spectrum of physiologic and immunologic functions, including endometrial receptivity and implantation as well as defense against sexually transmitted pathogens. Macrophages and epithelial cells cooperatively mediate innate host defense against bacterial invasion through the generation of immunologic effectors, including cytokines and antimicrobial peptides. In this study, we demonstrate that stimulation of peripheral blood monocytes and uterine macrophages with bacterial LPS induces the production of biologically active proinflammatory IL-1beta. High doses of estradiol enhance LPS-induced IL-1beta expression in an estrogen receptor dependent manner. Furthermore, both peripheral blood monocyte- and uterine macrophage-derived IL-1beta induce secretion of antimicrobial human beta-defensin 2 by uterine epithelial cells. These data indicate dynamic immunologic interaction between uterine macrophages and epithelial cells and implicate a role for estradiol in the modulation of the immune response. PMID- 16709824 TI - Macrophages induce neutrophil apoptosis through membrane TNF, a process amplified by Leishmania major. AB - Neutrophils are recruited to the site of parasite inoculation within a few hours of infection with the protozoan parasite Leishmania major. In C57BL/6 mice, which are resistant to infection, neutrophils are cleared from the site of s.c. infection within 3 days, whereas they persist for at least 10 days in susceptible BALB/c mice. In the present study, we investigated the role of macrophages (MPhi) in regulating neutrophil number. Inflammatory cells were recruited by i.p. injection of either 2% starch or L. major promastigotes. Neutrophils were isolated and cultured in the presence of increasing numbers of MPhi. Extent of neutrophil apoptosis positively correlated with the number of MPhi added. This process was strictly dependent on TNF because MPhi from TNF-deficient mice failed to induce neutrophil apoptosis. Assays using MPhi derived from membrane TNF knock in mice or cultures in Transwell chambers revealed that contact with MPhi was necessary to induce neutrophil apoptosis, a process requiring expression of membrane TNF. L. major was shown to exacerbate MPhi-induced apoptosis of neutrophils, but BALB/c MPhi were not as potent as C57BL/6 MPhi in this induction. Our results emphasize the importance of MPhi-induced neutrophil apoptosis, and membrane TNF in the early control of inflammation. PMID- 16709825 TI - An altered T cell repertoire in MECL-1-deficient mice. AB - Immunoproteasome subunits low-molecular mass polypeptide (LMP)2 and LMP7 affect Ag presentation by MHC class I molecules. In the present study, we investigated the function of the third immunosubunit LMP10/multicatalytic endopeptidase complex-like (MECL)-1 (beta2i) in MECL-1 gene-targeted mice. The number of CD8+ splenocytes in MECL-1-/- mice was 20% lower than in wild-type mice. Infection with lymphocytic choriomeningitis virus (LCMV) elicited a markedly reduced cytotoxic T cell (CTL) response to the LCMV epitopes GP276-286/Db and NP205 212/Kb in MECL-1-/- mice. The weak CTL response to GP276-286/Db was not due to an impaired generation of this epitope but was attributed to a decreased precursor frequency of GP276-286/Db-specific T cells. The expansion of TCR-Vbeta10+ T cells, which contain GP276-286/Db-specific cells, was reduced in LCMV-infected MECL-1-/- mice. Taken together, our data reveal an in vivo function of MECL-1 in codetermining the T cell repertoire for an antiviral CTL response. PMID- 16709826 TI - Clustering class I MHC modulates sensitivity of T cell recognition. AB - T cell recognition of peptide-MHC is highly specific and is sensitive to very low levels of agonist peptide; however, it is unclear how this effect is achieved or regulated. In this study we show that clustering class I MHC molecules on the cell surface of B lymphoblasts enhances their recognition by mouse and human T cells. We increased clustering of MHC I molecules by two methods, cholesterol depletion and direct cross-linking of a dimerizable MHC construct. Imaging showed that both treatments increased the size and intensity of MHC clusters on the cell surface. Enlarged clusters correlated with enhanced lysis and T cell effector function. Enhancements were peptide-specific and greatest at low concentrations of peptide. Clustering MHC class I enhanced recognition of both strong and weak agonists but not null peptide. Our results indicate that the lateral organization of MHC class I on the cell surface can modulate the sensitivity of T cell recognition of agonist peptide. PMID- 16709827 TI - Adhesion- and degranulation-promoting adapter protein is required for efficient thymocyte development and selection. AB - Adhesion- and degranulation-promoting adapter protein (ADAP) is required in TCR induced activation and proliferation of peripheral T cells. Loss of ADAP also impairs TCR-initiated inside-out activation of the integrin LFA-1 (CD11a/CD18, alphaLbeta2). In this study, we demonstrate that ADAP-deficient CD4/CD8 double positive (DP) cells have a diminished ability to proliferate, and that these DP thymocytes up-regulate CD69 poorly in vivo. Moreover, in both MHC class I- and class II-restricted TCR transgenic models, loss of ADAP interferes with both positive and negative selection. ADAP deficiency also impairs the ability of transgene-bearing DP thymocytes to form conjugates with Ag-loaded presenting cells. These findings suggest that ADAP is critical for thymocyte development and selection. PMID- 16709828 TI - Degeneracy and repertoire of the human HIV-1 Gag p17(77-85) CTL response. AB - CD8+ CTL responses are important for the control of HIV-1 infection. The immunodominant HLA-A2-restricted Gag epitope, SLYNTVATL (SL9), is considered to be a poor immunogen because reactivity to it is rare in acute infection despite its paradoxical dominance in patients with chronic infection. We have previously reported SL9 to be a help-independent epitope in that it primes highly activated CTLs ex vivo from CD8+ T cells of seronegative healthy donors. These CTLs produce sufficient cytokines for extended autocrine proliferation but are sensitive to activation-induced cell death, which may cause them to be eliminated by a proinflammatory cytokine storm. Here we identified an agonist variant of the SL9 peptide, p41 (SLYNTVAAL), by screening a large synthetic combinatorial nonapeptide library with ex vivo-primed SL9-specific T cells. p41 invariably immunized SL9-cross-reactive CTLs from other donors ex vivo and H-2Db beta2m double knockout mice expressing a chimeric HLA-A*0201/H2-Db MHC class I molecule. Parallel human T cell cultures showed p41-specific CTLs to be less fastidious than SL9-CTLs in the level of costimulation required from APCs and the need for exogenous IL-2 to proliferate (help dependent). TCR sequencing revealed that the same clonotype can develop into either help-independent or help-dependent CTLs depending on the peptide used to activate the precursor CD8+ T cells. Although Ag experienced SL9-T cells from two patients were also sensitive to IL-2-mediated cell death upon restimulation in vitro, the loss of SL9 T cells was minimized with p41. This study suggests that agonist sequences can replace aberrantly immunogenic native epitopes for the rational design of vaccines targeting HIV-1. PMID- 16709829 TI - IL-7R alpha gene expression is inversely correlated with cell cycle progression in IL-7-stimulated T lymphocytes. AB - IL-7 plays a major role in T lymphocyte homeostasis and has been proposed as an immune adjuvant for lymphopenic patients. This prospect is based, at least in part, on the short-term expansion of peripheral T cells in rIL7-treated mice and primates. Nevertheless, in vivo, following initial increases in T cell proliferation and numbers, lymphocytes return to a quiescent state. As the bases for this cell cycle exit have not yet been elucidated, it is important to assess the long-term biological effects of IL-7 on quiescent human T lymphocyte subsets. In this study, we find that IL-7-stimulated CD4+ naive lymphocytes enter into cell cycle with significantly delayed kinetics as compared with the memory population. Importantly though, these lymphocytes exit from the cell cycle despite the continuous replenishment of rIL-7. This response is distinct in memory and naive CD4+ lymphocytes with memory cells starting to exit from cycle by day 10 vs day 18 for naive cells. Return to quiescence is associated with a cessation in IL-7R signaling as demonstrated by an abrogation of STAT-5 phosphorylation, despite an up-regulation of surface IL-7Ralpha. Indeed, an initial 10-fold decrease in IL-7Ralpha mRNA levels is followed by increased IL 7Ralpha expression in naive as well as memory T cells, with kinetics paralleling cell cycle exit. Altogether, our data demonstrate that IL-7 promotes the extended survival of both naive and memory CD4+ T cells, whereas cycling of these two subsets is distinct and transient. Thus, IL-7 therapy should be designed to allow optimal responsiveness of naive and memory T cell subsets. PMID- 16709830 TI - Protein kinase C-theta-mediated signals enhance CD4+ T cell survival by up regulating Bcl-xL. AB - Productive engagement of TCR results in delivering signals required for T cell proliferation as well as T cell survival. Blocking TCR-mediated survival signals, T cells undergo apoptosis instead of proliferation upon TCR stimulation. During the activation process, T cells produce IL-2, which acts as an extrinsic survival factor. In addition, TCR stimulation results in up-regulation of Bcl-xL to enhance T cell survival intrinsically. We show in this study that protein kinase C (PKC)-theta is required for enhancing the survival of activated CD4+ T cells by up-regulating Bcl-xL. In response to TCR stimulation, CD4+ PKC-theta-/- T cells failed to up-regulate Bcl-xL, and underwent accelerated apoptosis via a caspase- and mitochondria-dependent pathway. Similar to PKC-theta-deficient primary CD4+ T cells, small interfering RNA-mediated knockdown of PKC-theta in Jurkat cells also resulted in apoptosis upon TCR stimulation. Forced expression of Bcl-xL was sufficient to inhibit apoptosis observed in PKC-theta knockdown cells. Furthermore, ectopic expression of PKC-theta stimulated a reporter gene driven by a mouse Bcl-xL promoter. Whereas an inactive form of PKC-theta or knockdown of endogenous PKC-theta led to inhibition of Bcl-xL reporter. PKC-theta-mediated activation of Bcl-xL reporter was inhibited by dominant-negative IkappaB kinase beta or dominant-negative AP-1. Thus, the PKC-theta-mediated signals may function not only in the initial activation of naive CD4+ T cells, but also in their survival during T cell activation by regulating Bcl-xL levels through NF-kappaB and AP-1 pathways. PMID- 16709831 TI - Lymphopenia-induced homeostatic proliferation of CD8+ T cells is a mechanism for effective allogeneic skin graft rejection following burn injury. AB - Burn patients are immunocompromised yet paradoxically are able to effectively reject allogeneic skin grafts. Failure to close a massive burn wound leads to sepsis and multiple system organ failure. Immune suppression early (3 days) after burn injury is associated with glucocorticoid-mediated T cell apoptosis and anti inflammatory cytokine responses. Using a mouse model of burn injury, we show CD8+ T cell hyperresponsiveness late (14 days) after burn injury. This is associated with a CD8+ T cell pro- and anti-inflammatory cytokine secretion profile, peripheral lymphopenia, and accumulation of a rapidly cycling, hyperresponsive memory-like CD8+CD44+ IL-7R- T cells which do not require costimulation for effective Ag response. Adoptive transfer of allospecific CD8+ T cells purified 14 days postburn results in enhanced allogeneic skin graft rejection in unburned recipient mice. Chemical blockade of glucocorticoid-induced lymphocyte apoptosis early after burn injury abolishes both the late homeostatic accumulation of CD8+ memory-like T cells and the associated enhanced proinflammatory CD8+ T cell response, but not the late enhanced CD8+ anti-inflammatory response. These data suggest a mechanism for the dynamic CD8+ T cell response following injury involving an interaction between activation, apoptosis, and cellular regeneration with broad clinical implications for allogeneic skin grafting and sepsis. PMID- 16709832 TI - CD86 stimulation on a B cell activates the phosphatidylinositol 3-kinase/Akt and phospholipase C gamma 2/protein kinase C alpha beta signaling pathways. AB - Stimulation of CD86 on a CD40L/IL-4-activated murine B cell increases the rate of mature IgG1 transcription by increasing the level of NF-kappaB activation, as well as Oct-2 expression and binding to the 3'-IgH enhancer. The signal transduction pathway activated by CD86 proximal to NF-kappaB activation is unknown. In this study, we show that CD86 stimulation on an activated B cell increases the activity of PI3K and the phosphorylation of phosphoinositide dependent kinase 1, Akt, and IkappaB kinase alphabeta. In addition, CD86 stimulation induces an increase in the phosphorylation of phospholipase Cgamma2 and protein kinase C alphabeta. CD86-mediated activation of these two signaling pathways leads to increased Oct-2 expression, increased gene activity mediated by NF-kappaB and 3'-IgH enhancer increased activity. These results identify a previously unknown signaling pathway induced by CD86 to regulate the level of B cell gene expression and activity. PMID- 16709833 TI - APRIL and BAFF promote increased viability of replicating human B2 cells via mechanism involving cyclooxygenase 2. AB - Of relevance to both protective and pathogenic responses to Ag is the recent finding that soluble molecules of the innate immune system, i.e., IL-4, B cell activation factor of the TNF family (BAFF), and C3, exhibit significant synergy in promoting the clonal expansion of human B2 cells following low-level BCR ligation. Although IL-4, BAFF, and C3dg each contribute to early cell cycle entry and progression to S phase, only BAFF promotes later sustained viability of progeny needed for continued cycling. The present study sought to further clarify the mechanisms for BAFF's multiple functions. By comparing BAFF and a proliferation-inducing ligand (APRIL) efficacy at different stages in the response (only BAFF binds BR3; both bind transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI) and B cell maturation Ag, the early role was attributed to BR3, while the later role was attributed to TACI/B cell maturation Ag. Importantly, BAFF- and APRIL-promoted viability of cycling lymphoblasts was associated with sustained expression of cyclooxygenase 2 (COX 2), the rate-limiting enzyme for PGE2 synthesis, within replicating cells. Supernatants of cultures with BAFF and APRIL contained elevated PGE2. Although COX-2 inhibitors diminished daughter cell viability, exogenous PGE2 (1-1000 nM) increased the viability and recovery of lymphoblasts. Increased yield of viable progeny was associated with elevated Mcl-1, suggesting that a BAFF/APRIL --> TACI --> COX-2 --> PGE2--> Mcl-1 pathway reduces activation-related, mitochondrial apoptosis in replicating human B2 cell clones. PMID- 16709834 TI - The combined effects of tryptophan starvation and tryptophan catabolites down regulate T cell receptor zeta-chain and induce a regulatory phenotype in naive T cells. AB - Tryptophan catabolism is a tolerogenic effector system in regulatory T cell function, yet the general mechanisms whereby tryptophan catabolism affects T cell responses remain unclear. We provide evidence that the short-term, combined effects of tryptophan deprivation and tryptophan catabolites result in GCN2 kinase-dependent down-regulation of the TCR zeta-chain in murine CD8+ T cells. TCR zeta down-regulation can be demonstrated in vivo and is associated with an impaired cytotoxic effector function in vitro. The longer-term effects of tryptophan catabolism include the emergence of a regulatory phenotype in naive CD4+CD25- T cells via TGF-beta induction of the forkhead transcription factor Foxp3. Such converted cells appear to be CD25+, CD69-, CD45RBlow, CD62L+, CTLA 4+, BTLAlow and GITR+, and are capable of effective control of diabetogenic T cells when transferred in vivo. Thus, both tryptophan starvation and tryptophan catabolites contribute to establishing a regulatory environment affecting CD8+ as well as CD4+ T cell function, and not only is tryptophan catabolism an effector mechanism of tolerance, but it also results in GCN2-dependent generation of autoimmune-preventive regulatory T cells. PMID- 16709836 TI - The mannose receptor mediates uptake of soluble but not of cell-associated antigen for cross-presentation. AB - The mannose receptor (MR) has been implicated in the recognition and clearance of microorganisms and serum glycoproteins. Its endocytic function has been studied extensively using macrophages, although it is expressed by a variety of cell types, including dendritic cells (DC). In this study, we investigated its role in Ag presentation by DC using MR-/- mice. Uptake of the model Ag, soluble OVA, by bone marrow-derived DC and in vitro activation of OVA-specific CD8 T cells (OT-I cells) strictly depended on the MR. In vivo, MR deficiency impaired endocytosis of soluble OVA by DC and concomitant OT-I cell activation. No alterations in the DC subtype composition in MR-/- mice were accountable. Uptake of cell-associated OVA was unaffected by MR deficiency, resulting in unchanged activation of OT-I cells. These findings demonstrate that DC use the MR for endocytosis of a particular Ag type intended for cross-presentation. PMID- 16709835 TI - Reduced KIR2DL1 recognition of MHC class I molecules presenting phosphorylated peptides. AB - As initially described by K. Karre and colleagues in the missing self hypothesis, cells expressing self-MHC class I proteins are protected from NK cells attack. In contrast, reduction in the expression of MHC class I molecules due to viral infection or tumor transformation result in the killing of these "abnormal" cells by NK cells via NK-activating receptors. Thus, NK killing of target cells is determined by both negative signals coming from MHC class I proteins and by positive signals derived from the activating ligands. The bound peptide in MHC class I play an important role in the balanced recognition of NK cells. The peptide stabilizes the MHC complex and interacts directly with the NK inhibitory receptors, thus participating in the determination of the fate of the target cells. In this study we demonstrate that posttranslational modifications such as phosphorylation of the presented peptide altered the ability of NK cells to recognize MHC class I molecules. By using a consensus peptide (QYDDAVYKL) that binds HLA-Cw4 in which different positions in the bound peptide were modified by serine phosphorylation, we observed a reduction in KIR2DL1 binding that led to decreased protection from NK killing. Therefore, it might be possible that alteration in the phosphorylation pattern during tumor transformation or viral infection may result in less inhibition and, consequently, improved NK cell killing. PMID- 16709837 TI - Deletion of exon I of SMAD7 in mice results in altered B cell responses. AB - The members of the TGF-beta superfamily, i.e., TGF-beta isoforms, activins, and bone morphogenetic proteins, regulate growth, differentiation, and apoptosis, both during embryonic development and during postnatal life. Smad7 is induced by the TGF-beta superfamily members and negatively modulates their signaling, thus acting in a negative, autocrine feedback manner. In addition, Smad7 is induced by other stimuli. Thus, it can fine-tune and integrate TGF-beta signaling with other signaling pathways. To investigate the functional role(s) of Smad7 in vivo, we generated mice deficient in exon I of Smad7, leading to a partial loss of Smad7 function. Mutant animals are viable, but significantly smaller on the outbred CD 1 mouse strain background. Mutant B cells showed an overactive TGF-beta signaling measured as increase of phosphorylated Smad2-positive B cells compared with B cells from wild-type mice. In agreement with this expected increase in TGF-beta signaling, several changes in B cell responses were observed. Mutant B cells exhibited increased Ig class switch recombination to IgA, significantly enhanced spontaneous apoptosis in B cells, and a markedly reduced proliferative response to LPS stimulation. Interestingly, LPS treatment reverted the apoptotic phenotype in the mutant cells. Taken together, the observed phenotype highlights a prominent role for Smad7 in development and in regulating the immune system's response to TGF-beta. PMID- 16709838 TI - Phosphatidylinositol 3-kinase/Akt positively regulates Fas (CD95)-mediated apoptosis in epidermal Cl41 cells. AB - Fas (CD95)-mediated apoptosis is an essential mechanism for the maintenance of homeostasis, and disruption of this death pathway contributes to many human diseases. The cell survival protein kinase Akt/protein kinase B (PKB) is a known regulator of apoptosis, but its role in Fas-mediated cell death and its regulatory mechanisms are unclear. In this study, we show that stimulation of the Fas receptor by its ligand (FasL) induces rapid phosphorylation of Akt/PKB and a parallel increase in cell apoptosis in epidermal Cl41 cells. Inhibition of PI3K/Akt by dominant-negative overexpression of PI3K (Deltap85) and Akt (Akt T308A/S473A) protects the cells from apoptosis, indicating an unexpected proapoptotic role of PI3K/Akt in the Fas signaling process. Treatment of the cells with pharmacological inhibitors of PI3K, wortmannin and 2-(4-morpholinyl)-8 phenyl-4H-1-benzopyran-4-1 (LY294002), similarly inhibits FasL-induced apoptosis and Akt/PKB phosphorylation, indicating that PI3K is an upstream mediator of Akt/PKB and is involved in Fas-mediated cell death. Electron spin resonance studies show that FasL treatment induces rapid generation of reactive oxygen species, and inhibition of ROS by antioxidants effectively inhibits Akt/PKB signaling, suggesting that FasL activation of Akt/PKB is redox sensitive. In cells transfected with dominant-negative PI3K/Akt, Fas expression is down regulated, but FLIP expression is unaffected. Reporter gene and mRNA expression assays show that FasL activates fas transcriptional activity and this effect is inhibited by PI3K/Akt suppression. Together, our results indicate that the PI3K/Akt, in addition to its normal prosurvival role, also plays an apoptotic role in Fas-mediated cell death through a mechanism that involves transcriptional activation of Fas receptor. PMID- 16709839 TI - Helicobacter pylori CagA-dependent macrophage migration inhibitory factor produced by gastric epithelial cells binds to CD74 and stimulates procarcinogenic events. AB - Macrophage migration inhibitory factor (MIF) is a proinflammatory cytokine that has recently been implicated in carcinogenesis. Helicobacter pylori, which is closely linked to gastric cancer, induces the gastric epithelium to produce proinflammatory cytokines, including MIF. MIF can bind to CD74, which we have previously shown to be highly expressed on the surface of gastric epithelial cells (GEC) during H. pylori infection. In this study, we sought to investigate the role of the H. pylori-induced MIF on epithelial proliferation and procarcinogenic events. Upon establishing a role for the H. pylori CagA virulence factor in MIF production, MIF binding to CD74 on GEC was confirmed. rMIF and H. pylori were shown to increase GEC proliferation, which was decreased when cagA- strains were used and when CD74 was blocked by mAbs. Apoptosis was also decreased by MIF, but increased by cagA- strains that induced much lower amounts of MIF than the wild-type bacteria. Furthermore, MIF binding to CD74 was also shown to decrease p53 phosphorylation and up-regulate Bcl-2 expression. This data describes a novel system in which an H. pylori virulence factor contributes to the production of a host factor that in turn up-regulates procarcinogenic events by the gastric epithelium. PMID- 16709840 TI - Central role for MyD88 in the responses of microglia to pathogen-associated molecular patterns. AB - Microglia, the innate immune effector cells of the CNS parenchyma, express TLR that recognize conserved motifs of microorganisms referred to as pathogen associated molecular patterns (PAMP). All TLRs identified to date, with the exception of TLR3, use a common adaptor protein, MyD88, to transduce activation signals. Recently, we reported that microglial activation in response to the Gram positive bacterium Staphylococcus aureus was not completely attenuated following TLR2 ablation, suggesting the involvement of additional receptors. To assess the functional role of alternative TLRs in microglial responses to S. aureus and its cell wall product peptidoglycan as well as the Gram-negative PAMP LPS, we evaluated primary microglia from MyD88 knockout (KO) and wild-type mice. The induction of TNF-alpha, IL-12 p40, and MIP-2 (CXCL2) expression by S. aureus- and peptidoglycan-stimulated microglia was MyD88 dependent, as revealed by the complete inhibition of cytokine production in MyD88 KO cells. In addition, the expression of additional pattern recognition receptors, including TLR9, pentraxin 3, and lectin-like oxidized LDL receptor-1, was regulated, in part, via a MyD88 dependent manner as demonstrated by the attenuated expression of these receptors in MyD88 KO microglia. Microglial activation was only partially inhibited in LPS stimulated MyD88 KO cells, suggesting the involvement of MyD88-independent pathways. Collectively, these findings reveal the complex mechanisms for microglia to respond to diverse bacterial pathogens, which occur via both MyD88 dependent and -independent pathways. PMID- 16709842 TI - A role for MAPK in feedback inhibition of Tcrb recombination. AB - The Tcrb locus is subject to a host of regulatory mechanisms that impart a strict cell and developmental stage-specific order to variable (V), diversity (D), and joining (J) gene segment recombination. The Tcrb locus is also regulated by allelic exclusion mechanisms, which restrict functional rearrangements to a single allele. The production of a functional rearrangement in CD4-CD8- double negative (DN) thymocytes leads to the assembly of a pre-TCR and initiates signaling cascades that allow for DN to CD4+CD8+ double-positive (DP) differentiation, proliferation, and feedback inhibition of further Vbeta to DJbeta rearrangement. Feedback inhibition is believed to be controlled, in part, by the loss of Vbeta gene segment accessibility during the DN to DP transition. However, the pre-TCR signaling pathways that lead to the inactivation of Vbeta chromatin have not been determined. Because activation of the MAPK pathway is documented to promote DP differentiation in the absence of allelic exclusion, we characterized the properties of Vbeta chromatin within DP thymocytes generated by a constitutively active Raf1 (Raf-CAAX) transgene. Consistent with previous reports, we show that the Raf-CAAX transgene does not inhibit Tcrb recombination in DN thymocytes. Nevertheless, DP thymocytes generated by Raf-CAAX signals display normal down-regulation of Vbeta segment accessibility and normal feedback inhibition of the Vbeta to DJbeta rearrangement. Therefore, our results emphasize the distinct requirements for feedback inhibition in the DN and DP compartments. Although MAPK activation cannot impose feedback in DN thymocytes, it contributes to feedback inhibition through developmental changes that are tightly linked to DN to DP differentiation. PMID- 16709841 TI - The TCR C beta FG loop regulates alpha beta T cell development. AB - The TCRbeta chain constant domain contains an unusually elongated, solvent exposed FG loop. This structural element forms one component of an alphabeta TCR cavity against which CD3epsilongamma may abut to facilitate Ag-specific signaling. Consistent with this notion, in the present study we show that N15alphabeta TCR transfectants expressing a FG loop-deleted chain (betaDeltaFG) stimulate less tyrosine protein phosphorylation than those bearing a wild-type beta-chain (betawt) upon TCR cross-linking. Furthermore, coimmunoprecipitation studies suggest a weakened association between the CD3epsilongamma heterodimer and the beta-chain in TCR complexes containing the betaDeltaFG variant. To further investigate the biologic role of the Cbeta FG loop in development, we competitively reconstituted the thymus of Ly5 congenic or RAG-2-/- mice using bone marrow cells from betawt or betaDeltaFG transgenic C57BL/6 (B6) mice. Both betawt and betaDeltaFG precursor cells generate thymocytes representative of all maturational stages. However, betaDeltaFG-expressing thymocytes dominate during subsequent development, resulting in an excess of betaDeltaFG-expressing peripheral T cells with reduced proliferative and cytokine production abilities upon TCR stimulation. Collectively, our results show that the unique Cbeta FG loop appendage primarily controls alphabeta T cell development through selection processes. PMID- 16709843 TI - Growth, differentiation, and malignant transformation of pre-B cells mediated by inducible activation of v-Abl oncogene. AB - The nonreceptor tyrosine kinase, encoded by the v-Abl oncogene of Abelson murine leukemia virus induces transformation of progenitor B cells. The v-Abl oncogene promotes cell cycle progression and inhibits pre-B cell differentiation. The temperature-sensitive form of Abelson murine leukemia virus offers a reversible model to study the role of v-Abl in regulating growth and differentiation. Inactivation of v-Abl elevates p27 and Foxo3a levels and activates NF-kappaB/Rel, which leads to G1 arrest and induction of Ig L chain gene rearrangement, respectively. In turn, v-Abl reactivation reduces p27 and Foxo3a levels, thus permitting G1-arrested cells to reenter the cell cycle. However, the cell lines derived from SCID mice that are defective in the catalytic subunit of DNA dependent protein kinase retain elevated levels of p27 and Foxo3a proteins despite reactivation of v-Abl. Consequently, these cells are locked in the G1 phase for an extended period of time. The few cells that manage to bypass the G1 arrest become tumorigenic and fail to undergo pre-B cell differentiation induced by v-Abl inactivation. Deregulation of p27, Foxo3a, c-myc, and NF-kappaB/Rel was found to be associated with the malignant transformation of SCID temperature sensitive form of Abelson murine leukemia virus pre-B cells. PMID- 16709844 TI - Identification of a candidate regulatory element within the 5' flanking region of the mouse Igh locus defined by pro-B cell-specific hypersensitivity associated with binding of PU.1, Pax5, and E2A. AB - The Igh locus is controlled by cis-acting elements, including Emu and the 3' IgH regulatory region which flank the C region genes within the well-studied 3' part of the locus. Although the presence of additional control elements has been postulated to regulate rearrangements of the VH gene array that extends to the 5' end of the locus, the 5' border of Igh and its flanking region have not been characterized. To facilitate the analysis of this unexplored region and to identify potential novel control elements, we physically mapped the most D-distal VH segments and scanned 46 kb of the immediate 5' flanking region for DNase I hypersensitive sites. Our studies revealed a cluster of hypersensitive sites 30 kb upstream of the most 5' VH gene. Detection of one site, HS1, is restricted to pro-B cell lines and HS1 is accessible to restriction enzyme digestion exclusively in normal pro-B cells, the stage defined by actively rearranging Igh V loci. Sequence motifs within HS1 for PU.1, Pax5, and E2A bind these proteins in vitro and these factors are recruited to HS1 sequence only in pro-B cells. Transient transfection assays indicate that the Pax5 binding site is required for the repression of transcriptional activity of HS1-containing constructs. Thus, our characterization of the region 5' of the VH gene cluster demonstrated the presence of a single cluster of DNase I hypersensitive sites within the 5' flanking region, and identified a candidate Igh regulatory region defined by pro B cell-specific hypersensitivity and interaction with factors implicated in regulating VDJ recombination. PMID- 16709847 TI - Evidence for a domain-swapped CD4 dimer as the coreceptor for binding to class II MHC. AB - CD4 is a coreceptor for binding of T cells to APC and the primary receptor for HIV. The disulfide bond in the second extracellular domain (D2) of CD4 is reduced on the cell surface, which leads to formation of disulfide-linked homodimers. A large conformational change must take place in D2 to allow for formation of the disulfide-linked dimer. Domain swapping of D2 is the most likely candidate for the conformational change leading to formation of two disulfide-bonds between Cys130 in one monomer and Cys159 in the other one. Mild reduction of the extracellular part of CD4 resulted in formation of disulfide-linked dimers, which supports the domain-swapped model. The functional significance of dimer formation for coreceptor function was tested using cells expressing wild-type or disulfide bond mutant CD4. Eliminating the D2 disulfide bond markedly impaired CD4's coreceptor function. Modeling of the complex of the TCR and domain-swapped CD4 dimer bound to class II MHC and Ag supports the domain-swapped dimer as the immune coreceptor. The known involvement of D4 residues Lys318 and Gln344 in dimer formation is also accommodated by this model. These findings imply that disulfide-linked dimeric CD4 is the preferred coreceptor for binding to APC. PMID- 16709845 TI - The Fas-associated death domain protein is required in apoptosis and TLR-induced proliferative responses in B cells. AB - The Fas-associated death domain protein (FADD)/Mort1 is a signaling adaptor protein which mediates the activation of caspase 8 during death receptor-induced apoptosis. Disruption of FADD in germ cells results in death receptor-independent embryonic lethality in mice. Previous studies indicated that in addition to its function in apoptosis, FADD is also required in peripheral T cell homeostasis and TCR-induced proliferative responses. In this report, we generated B cell-specific FADD-deficient mice and showed that deletion of FADD at the pro-B cell stage had minor effects on B cell development in the bone marrow, and resulted in increased splenic and lymph node B cell numbers and decreased peritoneal B1 cell numbers. As in T cells, a FADD deficiency inhibited Fas-induced apoptosis in B cells. However, B cell-proliferative responses induced by stimulation of the BCR and CD40 using anti-IgM or anti-CD40 Abs were unaffected by the absence of FADD. Further analyses revealed that FADD-deficient B cells were defective in proliferative responses induced by treatments with dsRNA and LPS which stimulate TLR3 and TLR4, respectively. Therefore, in addition to its apoptotic function, FADD also plays a role in TLR3- and TLR4-induced proliferative responses in B cells. PMID- 16709846 TI - Precursor B cell receptor signaling activity can be uncoupled from surface expression. AB - Signals from the precursor BCR (preBCR) cause proliferation and differentiation of progenitor (pro-) B cells into pre-B cells. Given the very low amounts of surface preBCRs and the demonstrated cell autonomy of preBCR signaling, we examined the possible occurrence of preBCR signal propagation from intracellular membranes such as the endoplasmic reticulum (ER) and the trans-Golgi network (TGN) in transformed and primary pro-B cells. PreBCRs composed of normal Ig mu or truncated Dmu heavy chains (HCs) were redirected to intracellular sites via localization sequences appended to the HC cytoplasmic tail. PreBCR complexes retained in the TGN or shunted from the TGN to lysosomes were as or 50% as active as the corresponding wild-type preBCRs in directing preBCR-dependent events, including CD2 and CD22 expression and proliferation in primary pro-B cells. This occurred despite their low to undetectable surface expression in transformed cells, which otherwise allowed significant surface accumulation of wild-type preBCRs. In contrast, ER-retained preBCRs were inactive. These results suggest that preBCR signaling is remarkably tolerant of dramatic changes in its subcellular distribution within post-ER compartments and support the possibility that the preBCR can activate signaling pathways in the TGN as well as the plasma membrane. PMID- 16709848 TI - Development and selection of edited B cells in B6.56R mice. AB - Tolerance to dsDNA is broken in mice with a high-affinity anti-DNA H chain transgene, 56R, on the C57BL/6 background (B6.56R). B6.56R produce more anti dsDNA Abs than BALBc.56R. To investigate how anti-DNA Abs are regulated on the B6 background, phenotypic and genetic studies were performed. B6.56R have reduced numbers of B cells and phenotypically altered B cell subsets, including relative increases in the proportions of IgM-negative bone marrow B cells, cells with a marginal zone phenotype, and cells with a transitional T3 phenotype. The peripheral B cell repertoire in B6.56R is restricted: most B cells express the 56R H chain and use a similar, limited subset of editor L chains. DNA binding is more common in B6.56R because the repertoire is shifted toward L chains that are more permissive for DNA binding. H chain editing is also observed and is increased in spontaneous as compared with LPS hybridomas. A subset of spontaneous hybridomas appears to lack H chain expression. PMID- 16709849 TI - Immunologic consequences of Francisella tularensis live vaccine strain infection: role of the innate immune response in infection and immunity. AB - Francisella tularensis (Ft), a Gram-negative intracellular bacterium, is the etiologic agent of tularemia. Although attenuated for humans, i.p. infection of mice with <10 Ft live vaccine strain (LVS) organisms causes lethal infection that resembles human tularemia, whereas the LD50 for an intradermal infection is >10(6) organisms. To examine the immunological consequences of Ft LVS infection on the innate immune response, the inflammatory responses of mice infected i.p. or intradermally were compared. Mice infected i.p. displayed greater bacterial burden and increased expression of proinflammatory genes, particularly in the liver. In contrast to most LPS, highly purified Ft LVS LPS (10 microg/ml) was found to be only minimally stimulatory in primary murine macrophages and in HEK293T cells transiently transfected with TLR4/MD-2/CD14, whereas live Ft LVS bacteria were highly stimulatory for macrophages and TLR2-expressing HEK293T cells. Despite the poor stimulatory activity of Ft LVS LPS in vitro, administration of 100 ng of Ft LVS LPS 2 days before Ft LVS challenge severely limited both bacterial burden and cytokine mRNA and protein expression in the absence of detectable Ab at the time of bacterial challenge, yet these mice developed a robust IgM Ab response within 2 days of infection and survived. These data suggest that prior administration of Ft LVS LPS protects the host by diminishing bacterial burden and blunting an otherwise overwhelming inflammatory response, while priming the adaptive immune response for development of a strong Ab response. PMID- 16709850 TI - HIV-1 adapts to a retrocyclin with cationic amino acid substitutions that reduce fusion efficiency of gp41. AB - Retrocyclin (RC)-101 is a cationic theta-defensin that inhibits HIV-1 entry. Passaging HIV-1(BAL) under selective pressure by this cyclic minidefensin resulted in only a 5- to 10-fold decrease in viral susceptibility to RC-101. Emergent viral isolates had three amino acid substitutions in their envelope glycoprotein. One was in a CD4-binding region of gp120, and the others were in the heptad repeat (HR) domains of gp41 (HR1 and HR2). Each mutation replaced an electroneutral or electronegative residue with one that was positively charged. These mutations were evaluated either alone or in combination in a single-round viral entry assay. Although the mutation in gp120 did not affect viral entry, the mutation in HR1 of gp41 conferred relative resistance to RC-101. Interestingly, the envelope with the HR2 mutation was less efficient and became codependent on the presence of RC-101 for entry. The adaptive response of HIV-1 to this cationic host defense peptide resembles the responses of bacteria that modulate their surface or membrane charge to evade analogous host defense peptides. These findings also suggest that interactions between theta-defensins and gp41 may contribute to the ability of these cyclic minidefensins to prevent HIV-1 entry into target cells. PMID- 16709851 TI - PU.1 and a TTTAAA element in the myeloid defensin-1 promoter create an operational TATA box that can impose cell specificity onto TFIID function. AB - Defensins are major components of a peptide-based, antimicrobial system in human neutrophils. While packed with peptide, circulating cells contain no defensin-1 (def1) transcripts, except in some leukemia patients and in derivative promyelocytic leukemia cell lines. Expression is modulated by serum factors, mediators of inflammation, and kinase activators and inhibitors, but the underlying mechanisms are not fully understood. A minimal def1 promoter drives transcription in HL-60 cells under control of PU.1 and a def1-binding protein ("D1BP"), acting through, respectively, proximal (-22/-19) and distal (-62/-59) GGAA elements. In this study, we identify D1BP, biochemically and functionally, as GA-binding protein (GABP)alpha/GABPbeta. Whereas GABP operates as an essential upstream activator, PU.1 assists the flanking "TTTAAA" element (-32/-27), a "weak" but essential TATA box, to bring TBP/TFIID to the transcription start site. PU.1 thus imparts a degree of cell specificity to the minimal promoter and provides a potential link between a number of signaling pathways and TFIID. However, a "strong" TATA box ("TATAAA") eliminates the need for the PU.1 binding site and for PU.1, but not for GABP. As GABP is widely expressed, a strong TATA box thus alleviates promyelocytic cell specificity of the def1 promoter. These findings suggest how the myeloid def1 promoter may have evolutionarily acquired its current properties. PMID- 16709852 TI - F4/80+ alternatively activated macrophages control CD4+ T cell hyporesponsiveness at sites peripheral to filarial infection. AB - Both T cells and APC have been strongly implicated in the immune suppression observed during filarial nematode infections, but their relative roles are poorly understood, particularly in regard to timing and locality of action. Using Litomosoides sigmodontis infection of susceptible BALB/c mice, we have studied the progression of filarial immunosuppression leading to patent infection with blood microfilaremia. Patent infection is associated with decreased immune responsiveness in the draining thoracic lymph nodes (tLN) and intrinsically hyporesponsive CD4+ T cells at the infection site. We now show that we are able to separate, both in time and space, different suppressive mechanisms and cell populations that contribute to filarial hyporesponsiveness. L. sigmodontis infection recruited a F4/80+ population of alternatively activated macrophages that potently inhibited Ag-specific CD4+ T cell proliferative responses even in the presence of competent naive APC. T cell responsiveness was partially restored by neutralizing TGF-beta, but not by blocking IL-10 or CTLA-4 signaling. During prepatent infection, the macrophage population was restricted to the infection site. However, once infection became patent with systemic release of microfilariae, the suppressive macrophage activity extended peripherally into the tLN. In contrast, the hyporesponsive CD4+ T cell phenotype remained localized at the infection site, and the tLN CD4+ T cell population recovered full Ag responsiveness in the absence of suppressive macrophages. Filarial immunosuppression, therefore, evolves over time at sites increasingly distal to infection, and the mechanisms of filarial down-regulation are dependent on proximity to the infection site. PMID- 16709853 TI - Epitopes derived by incidental translational frameshifting give rise to a protective CTL response. AB - Aberrant gene expression can be caused by several different mechanisms at the transcriptional, RNA processing, and translational level. Although most of the resulting proteins may have no significant biological function, they can be meaningful for the immune system, which is sensitive to extremely low levels of Ag. We have tested this possibility by investigating the ability of CD8+ T cells (TCD8+) to respond to an epitope whose expression results from incidental ribosomal frameshifting at a sequence element within the HSV thymidine kinase gene. This element, with no apparent functional significance, has been identified due to its ability to facilitate escape from the antiviral compound acyclovir. Using a recombinant vaccinia virus expression system, we find that in vitro and in vivo TCD8+ responses to the frameshift-dependent epitope are easily discernible. Furthermore, the in vivo response is at a sufficient level to mediate protection from a tumor challenge. Thus, the targets of immune responses to infectious agents can extend beyond the products of conventional open reading frames. On a per-cell basis, responses to such minimally expressed epitopes may be exceedingly effective due to the selective expansion of high avidity TCD8+. PMID- 16709854 TI - Identification of HLA-A*0201-presented T cell epitopes derived from the oncofetal antigen-immature laminin receptor protein in patients with hematological malignancies. AB - The oncofetal Ag immature laminin receptor (OFA-iLR) is a potential target molecule for immunotherapeutic studies in several tumor entities, including hematological malignancies. In the present study, we characterize two HLA-A*0201 presented epitopes eliciting strong OFA-iLR peptide-specific human cytotoxic T cell (CTLs) responses in vitro. Both allogeneic HLA-A*0201-matched and autologous CTLs recognized and killed endogenously OFA-iLR-expressing tumor cell lines and primary malignant cells from patients with hemopoietic malignancies in an MHC restricted fashion but spared nonmalignant hemopoietic cells. Spontaneous OFA-iLR peptide-specific T cell reactivity was detectable in a significant proportion of leukemia patients. Interestingly, in patients with chronic lymphocytic leukemia and multiple myeloma but not in those with acute myeloid leukemia, significant frequencies of OFA peptide-specific CTLs could be detected in an early stage of disease but disappeared in patients with progressive disease. The identification of OFA-iLR-derived peptide epitopes provides a basis for tumor immunological studies and therapeutic vaccination strategies in patients with OFA-iLR expressing malignancies. PMID- 16709855 TI - Mature hepatocyte growth factor/scatter factor on the surface of human granulocytes is released by a mechanism involving activated factor Xa. AB - Serum hepatocyte growth factor (HGF) is rapidly increased in patients suffering from various tissue injuries including arterial occlusive diseases. However, the cellular sources of the HGF increase remain largely unknown. In the present study, we showed that bioactive mature HGF is constitutively present on the surface of granulocytes in human peripheral blood. Exogenously added 125I-labeled iodo-HGF efficiently bound to granulocyte surface, whereas only a scarce amount of HGF mRNA was detected in granulocytes, indicating that the mature HGF on granulocytes is likely to be derived from other cell types. Interestingly, treatment of granulocytes with human serum rapidly induced the release of the cell surface-associated HGF. In vivo, thromboplastin injection into mice increased HGF release from transplanted human granulocytes, which was inhibited by the pretreatment with DX9065a, a specific inhibitor of factor Xa. Furthermore, DX9065a also inhibited the serum-induced HGF release from human granulocytes in vitro, suggesting that the HGF-releasing factor(s) in serum is associated with factor Xa activation. Thus, human granulocytes may function as a transporter of HGF in the peripheral blood, releasing HGF at the injured sites caused by blood coagulation, where HGF may promote tissue repair. PMID- 16709856 TI - Sequencing, expression, and functional analyses support the candidacy of Ncf2 in susceptibility to Salmonella typhimurium infection in wild-derived mice. AB - A recessive Salmonella Typhimurium susceptibility locus (immunity to Typhimurium (Ity3) was reported previously on distal mouse chromosome 1 using a cross between C57BL/6J and wild-derived MOLF/Ei mice. This quantitative trait locus is located in a genomic region spanning 84 Mb, rich in candidate genes for which a role in host resistance to Salmonella infection is either known or can be envisioned. In this study, we report the evaluation of neutrophil cytosolic factor 2 (Ncf2) as a candidate Salmonella susceptibility gene for Ity3. Ncf2 encodes p67phox, a subunit of the multiprotein enzyme complex NADPH oxidase, known to be responsible for the generation of superoxides. Congenic mice carrying the Ity3 region from MOLF/Ei, B6.MOLF-Ity/Ity3 were more susceptible to infection compared with control mice heterozygous at Ity3, B6.MOLF-Ity/Ity3(MOLF/B6), confirming the existence of a recessive Salmonella susceptibility locus on distal chromosome 1. Spleen Ncf2 expression levels were lower in infected congenic mice homozygous for the MOLF/Ei allele at Ity3 compared with mice heterozygous at Ity3. C57BL/6J and MOLF/Ei Ncf2 sequence comparisons revealed one nonconservative amino acid change (R394Q) in the functional and highly conserved Phox and Bem1 domain of the protein. Functional analysis revealed that the MOLF/Ei allele had reduced PMA- and Salmonella-induced superoxide induction as compared with their wild-type counterparts ex vivo. The R394Q substitution seems to occur on an amino acid involved in electrostatic interactions with p40phox, crucial in its activation. Moreover, a human mutation in the corresponding R395W, resulting in chronic granulatomous disease, is known to lead to reduced superoxide levels. These results support the candidacy of Ncf2 as the gene underlying Ity3. PMID- 16709857 TI - Innate defense against influenza A virus: activity of human neutrophil defensins and interactions of defensins with surfactant protein D. AB - Surfactant protein D (SP-D) plays important roles in innate host defense against influenza A virus (IAV) infection, in part by modifying interactions with neutrophils. Human neutrophil defensins (HNPs) inhibit infectivity of enveloped viruses, including IAV. Our goal in this study was to characterize antiviral interactions between SP-D and HNPs. Recombinant and/or natural forms of SP-D and related collectins and HNPs were tested for antiviral activity against two different strains of IAV. HNPs 1 and 2 did not inhibit viral hemagglutination activity, but they interfered with the hemagglutination-inhibiting activity of SP D. HNPs had significant viral neutralizing activity against divergent IAV strains. However, the HNPs generally had competitive effects when combined with SP-D in assays using an SP-D-sensitive IAV strain. In contrast, cooperative antiviral effects were noted in some instances when relatively SP-D-resistant strains were treated with SP-D and HNPs. HNPs were found to bind to the neck and/or carbohydrate recognition domain of SP-D. This binding was specific because no, or minimal, binding to other collectins was found. HNPs precipitated SP-D from bronchoalveolar lavage fluid and reduced the antiviral activity of bronchoalveolar lavage fluid. HNP-1 and -2 differed somewhat in their independent antiviral activity and their binding to SP-D. These results are relevant to the early phase of host defense against IAV, and suggest a complex interplay between SP-D and HNPs at sites of active inflammation. PMID- 16709858 TI - Maintenance of HIV-specific CD4+ T cell help distinguishes HIV-2 from HIV-1 infection. AB - Unlike HIV-1-infected people, most HIV-2-infected subjects maintain a healthy CD4+ T cell count and a strong HIV-specific CD4+ T cell response. To define the cellular immunological correlates of good prognosis in HIV-2 infection, we conducted a cross-sectional study of HIV Gag-specific T cell function in HIV-1- and HIV-2-infected Gambians. Using cytokine flow cytometry and lymphoproliferation assays, we show that HIV-specific CD4+ T cells from HIV-2 infected individuals maintained proliferative capacity, were not terminally differentiated (CD57-), and more frequently produced IFN-gamma or IL-2 than CD4+ T cells from HIV-1-infected donors. Polyfunctional (IFN-gamma+/IL-2+) HIV specific CD4+ T cells were found exclusively in HIV-2+ donors. The disparity in CD4+ T cell responses between asymptomatic HIV-1- and HIV-2-infected subjects was not associated with differences in the proliferative capacity of HIV-specific CD8+ T cells. This study demonstrates that HIV-2-infected donors have a well preserved and functionally heterogeneous HIV-specific memory CD4+ T cell response that is associated with delayed disease progression in the majority of infected people. PMID- 16709859 TI - STAT1 regulates IFN-alpha beta- and IFN-gamma-dependent control of infection with Chlamydia pneumoniae by nonhemopoietic cells. AB - STAT1 mediates signaling in response to IFN-alpha, -beta, and -gamma, cytokines required for protective immunity against several viral, bacterial, and eukaryotic pathogens. The protective role of STAT1 in the control of intranasal infection with the obligate intracellular bacterium Chlamydia pneumoniae was analyzed. IFN gamma-/- or IFN-gamma receptor (R)-/- mice were highly susceptible to infection with C. pneumoniae. We found that STAT1-/- mice were even more susceptible to C. pneumoniae than IFN-gamma-/- or IFN-gammaR-/- mice. Phosphorylation of STAT1 was detected in the lungs of C. pneumoniae-infected wild-type, IFN-gammaR-/-, and IFN alphabetaR-/- mice, but not in mice lacking both IFN-alphabetaR and IFN-gammaR. In line with this, IFN-alphabetaR-/-/IFN-gammaR-/- mice showed increased susceptibility to infection compared with IFN-gammaR-/- mice. However, C. pneumoniae-infected IFN-alphabetaR-/- or IFN regulatory factor 3-/- mice showed no increased susceptibility and similar IFN-gamma expression compared with wild type mice. CD4+ or CD8+ cells released IFN-gamma in vivo and conferred protection against C. pneumoniae in a STAT1-independent manner. In contrast, STAT1 mediated a nonredundant protective role of nonhemopoietic cells but not of hemopoietic cells. Nonhemopoietic cells accounted for the expression of STAT1-mediated indoleamine 2, 3-dioxygenase and the p47 GTPase LRG-47, but not inducible NO synthase mRNA. In summary, we demonstrate that STAT1 mediates a cooperative effect of IFN-alphabeta and IFN-gamma on nonhemopoietic cells, resulting in protection against C. pneumoniae. PMID- 16709860 TI - Leukocyte recruitment to the inflamed glomerulus: a critical role for platelet derived P-selectin in the absence of rolling. AB - The renal glomerulus is one of the few sites within the microvasculature in which leukocyte recruitment occurs in capillaries. However, due to the difficulty of directly visualizing the glomerulus, the mechanisms of leukocyte recruitment to glomerular capillaries are poorly understood. To overcome this, we rendered murine kidneys hydronephrotic to allow the visualization of the functional glomerular microvasculature during an inflammatory response. These experiments demonstrated that following infusion of anti-glomerular basement membrane (GBM) Ab, leukocytes became adherent in glomerular capillaries via a process of immediate arrest, without undergoing prior detectable rolling. However, despite the absence of rolling, this recruitment involved nonredundant roles for the P selectin/P-selectin glycoprotein ligand-1 and beta2 integrin/ICAM-1 pathways, suggesting that a novel form of the multistep leukocyte adhesion cascade occurs in these vessels. Anti-GBM Ab also increased glomerular P-selectin expression and induced a P-selectin-independent increase in platelet accumulation. Moreover, platelet depletion prevented both the increase in glomerular P-selectin, and the leukocyte recruitment induced by anti-GBM Ab. Furthermore, depletion of neutrophils and platelets also prevented the increase in urinary protein excretion induced by anti-GBM Ab, indicating that their accumulation in glomeruli contributed to the development of renal injury. Finally, infusion of wild-type platelets into P-selectin-deficient mice restored the ability of glomeruli in these mice to support leukocyte adhesion. Together, these data indicate that anti GBM Ab-induced leukocyte adhesion in glomeruli occurs via a novel pathway involving a nonrolling interaction mediated by platelet-derived P-selectin. PMID- 16709861 TI - Antifibrotic activity of an inhibitor of group IIA secretory phospholipase A2 in young spontaneously hypertensive rats. AB - The development of fibrosis in the chronically hypertensive heart is associated with infiltration of inflammatory cells and cardiac hypertrophy. In this study, an inhibitor of the proinflammatory enzyme, group IIA human secretory phospholipase A2 (sPLA2-IIA), has been found to prevent collagen deposition as an important component of cardiovascular remodeling in a rat model of developing chronic hypertension. Daily treatment of young male spontaneously hypertensive rats (SHR) with an sPLA2-IIA inhibitor (KH064, 5-(4-benzyloxyphenyl)-4S-(phenyl heptanoylamino)-pentanoic acid, 5 mg/kg/day p.o.) prevented increases in the content of perivascular (SHR 20.6 +/- 0.9%, n = 5; SHR+KH064 14.0 +/- 1.2%, n = 5) and interstitial (SHR 7.9 +/- 0.3%, n = 6; SHR+KH064 5.4 +/- 0.7%, n = 6) collagen in the left ventricle of rat hearts, but did not affect numbers of infiltrating monocytes/macrophages, left ventricular hypertrophy (SHR 2.88 +/- 0.08, n = 12; SHR+KH064 3.09 +/- 0.08 mg/g body weight, n = 9), increased systolic blood pressure, or thoracic aortic responses. This selective antifibrotic activity suggests that sPLA2-IIA may have an important but specific role in cardiac fibrosis, and that its inhibitors could be useful in dissecting molecular pathways leading to fibrotic conditions. PMID- 16709862 TI - Different stabilities of the structurally related receptors for IgE and IgG on the cell surface are determined by length of the stalk region in their alpha chains. AB - A variant of the high affinity IgE receptor FcepsilonRI, which is composed of alpha- and gamma-chains without the beta-chain, is expressed on human APC, such as dendritic cells, and has been suggested to facilitate Ag uptake through IgE and hence to facilitate Ag presentation to T cells. The level of FcepsilonRI on these cells is correlated with the serum IgE concentration, suggesting IgE mediates the up-regulation of the alphagamma2-type FcepsilonRI. The IgE-mediated FcepsilonRI up-regulation on mast cells and basophils has been shown to enhance the ability of these cells to release chemical mediators and cytokines that are responsible for allergic inflammatory reactions. Here, to elucidate the mechanism controlling FcepsilonRI expression, we compared two structurally related Ig receptors, human FcepsilonRI and FcgammaRIIIA, which carry different alpha-chains but the same gamma-chains. The half-life of FcepsilonRI on the cell surface was short unless it bound IgE, whereas FcgammaRIIIA was stably expressed without IgG binding. Shuffling of the non Ig-binding portions of the FcepsilonRIalpha and FcgammaRIIIAalpha chains revealed that the stalk region was critical in determining the difference in their stability and ligand-induced up-regulation. Unexpectedly, analyses with added or deleted amino acids in the stalk region strongly suggested that the length rather than the amino acid sequence of the stalk region was of major importance in determining the different stabilities of FcepsilonRI and FcgammaRIIIA on the cell surface. This finding provides new insights into the mechanism regulating surface FcepsilonRI expression. PMID- 16709863 TI - Decreased capacity of immune cells to cause tissue injury mediates kidney ischemic preconditioning. AB - Ischemic preconditioning (IP) is a well-established phenomenon, and the underlying mechanisms of IP are thought to involve adaptive changes within the injured tissue. Because one of the main functions of immune cells is to harbor memory, we hypothesized that circulating immune cells could mediate IP by responding to an initial ischemia reperfusion injury (IRI) and then mediate decreased injury after a second IRI event. C57BL/6 mice underwent 30 min of bilateral renal clamping or sham operation. At 5 days after ischemia, purified leukocytes from spleen were adoptively transferred into T cell-deficient (nu/nu) mice. After 1 wk, these mice underwent 30 min of renal IRI. The nu/nu mice receiving leukocytes from ischemic wild-type mice had significantly reduced renal injury compared with nu/nu mice receiving leukocytes from sham-operated, wild type mice. Infiltration of neutrophil and macrophage in postischemic kidney did not correlate with the protection. No difference in kidney C3d or IgG deposition was detected between groups. Given that inducible NO synthase (iNOS) has been implicated in IP, leukocytes from ischemic or sham-operated, iNOS-deficient mice were transferred into nu/nu mice. Effects similar to those of wild-type transfer of ischemic leukocytes were demonstrated; thus, iNOS was not mediating the IP effect of leukocytes. This is the first evidence that immune cells are primed after renal IRI and thereby lose the capacity to cause kidney injury during a second episode of IRI. This finding may also be relevant for elucidating the mechanisms underlying cross-talk between injured kidney and distant organs. PMID- 16709864 TI - Identification of small heat shock protein B8 (HSP22) as a novel TLR4 ligand and potential involvement in the pathogenesis of rheumatoid arthritis. AB - Dendritic cells (DCs) are specialized APCs that can be activated upon pathogen recognition as well as recognition of endogenous ligands, which are released during inflammation and cell stress. The recognition of exogenous and endogenous ligands depends on TLRs, which are abundantly expressed in synovial tissue from rheumatoid arthritis (RA) patients. Furthermore TLR ligands are found to be present in RA serum and synovial fluid and are significantly increased, compared with serum and synovial fluid from healthy volunteers and patients with systemic sclerosis and systemic lupus erythematosus. Identification of novel endogenous TLR ligands might contribute to the elucidation of the role of TLRs in RA and other autoimmune diseases. In this study, we investigated whether five members of the small heat shock protein (HSP) family were involved in TLR4-mediated DC activation and whether these small HSPs were present in RA synovial tissue. In vitro, monocyte-derived DCs were stimulated with recombinant alphaA crystallin, alphaB crystallin, HSP20, HSPB8, and HSP27. Using flow cytometry and multiplex cytokine assays, we showed that both alphaA crystallin and HSPB8 were able to activate DCs and that this activation was TLR4 dependent. Furthermore, Western blot and immunohistochemistry showed that HSPB8 was abundantly expressed in synovial tissue from patients with RA. With these experiments, we identified sHSP alphaA crystallin and HSPB8 as two new endogenous TLR4 ligands from which HSPB8 is abundantly expressed in RA synovial tissue. These findings suggest a role for HSPB8 during the inflammatory process in autoimmune diseases such as RA. PMID- 16709865 TI - Cytokine-induced hepatic apoptosis is dependent on FGL2/fibroleukin: the role of Sp1/Sp3 and STAT1/PU.1 composite cis elements. AB - Previous studies from our laboratory have shown that fulminant hepatitis caused by the mouse hepatitis virus, MHV-3, is dependent on production of the novel immune coagulant fgl2/fibroleukin. In this study, we investigate the role of IFN gamma and TNF-alpha in the induction of fgl2 expression and fgl2-dependent hepatic apoptosis. Infusion of IFN-gamma in combination with TNF-alpha through the portal vein of fgl2+/+ mice led to widespread hepatic apoptosis and fibrin deposition. Livers from fgl2-/- mice were normal, although strong expression of the fgl2 knockout reporter gene Lac Z was seen in both resident hepatic macrophages and endothelial cells. In vitro, IFN-gamma and TNF-alpha induced fgl2 expression in a macrophage and endothelial cell-specific manner. In macrophages (peritoneal and RAW 264.7 cells), IFN-gamma, but not IFN-alpha, LPS, TNF-alpha, or IL-1 induced fgl2 mRNA transcription and protein expression, while in endothelial cells TNF-alpha, but not IFN-gamma, induced fgl2 transcription. In addition, while TNF-alpha enhanced IFN-gamma-induced macrophage fgl2 transcription, IFN-gamma also enhanced TNF-alpha-induced endothelial cell fgl2 transcription. The induction of fgl2 by IFN-gamma in macrophages involved a STAT1 dependent pathway, involving the composite cis elements Sp1/Sp3 and GAS/PU.1. The latter interacted with IFN-gamma-dependent Sp1/Sp3, STAT1, and the ETS family of transcription factors member PU.1. The interaction of PU.1 with the IFN-gamma activated sequence/ETS family of transcription factors site determined the macrophage-specific induction of fgl2 by IFN-gamma. Overall, this study demonstrates that IFN-gamma and TNF-alpha induce hepatocyte apoptosis in vivo, which is dependent on induction of fgl2, and defines the molecular basis of transcription of fgl2 in vitro. PMID- 16709866 TI - Platelet-activating factor-induced clathrin-mediated endocytosis requires beta arrestin-1 recruitment and activation of the p38 MAPK signalosome at the plasma membrane for actin bundle formation. AB - Clathrin-mediated endocytosis (CME) is a common pathway used by G protein-linked receptors to transduce extracellular signals. We hypothesize that platelet activating factor (PAF) receptor (PAFR) ligation requires CME and causes engagement of beta-arrestin-1 and recruitment of a p38 MAPK signalosome that elicits distinct actin rearrangement at the receptor before endosomal scission. Polymorphonuclear neutrophils were stimulated with buffer or 2 microM PAF (1 min), and whole cell lysates or subcellular fractions were immunoprecipitated or slides prepared for colocalization and fluorescent resonance energy transfer analysis. In select experiments, beta-arrestin-1 or dynamin-2 were neutralized by intracellular introduction of specific Abs. PAFR ligation caused 1) coprecipitation of the PAFR and clathrin with beta-arrestin-1, 2) fluorescent resonance energy transfer-positive interactions among the PAFR, beta-arrestin-1, and clathrin, 3) recruitment and activation of the apoptosis signal-regulating kinase-1/MAPK kinase-3/p38 MAPK (ASK1/MKK3/p38 MAPK) signalosome, 4) cell polarization, and 5) distinct actin bundle formation at the PAFR. Neutralization of beta-arrestin-1 inhibited all of these cellular events, including PAFR internalization; conversely, dynamin-2 inhibition only affected receptor internalization. Selective p38 MAPK inhibition globally abrogated actin rearrangement; however, inhibition of MAPK-activated protein kinase-2 and its downstream kinase leukocyte-specific protein-1 inhibited only actin bundle formation and PAFR internalization. In addition, ASK1/MKK3/p38 MAPK signalosome assembly appears to occur in a novel manner such that the ASK1/p38 MAPK heterodimer is recruited to a beta-arrestin-1 bound MKK3. In polymorphonuclear neutrophils, leukocyte-specific protein-1 may play a role similar to fascin for actin bundle formation. We conclude that PAF signaling requires CME, beta arrestin-1 recruitment of a p38 MAPK signalosome, and specific actin bundle formation at the PAFR for transduction before endosomal scission. PMID- 16709867 TI - Leukotriene A4 hydrolase expression in PEL cells is regulated at the transcriptional level and leads to increased leukotriene B4 production. AB - Primary effusion lymphoma (PEL) is a herpesvirus-8-associated lymphoproliferative disease characterized by migration of tumor cells to serous body cavities. PEL cells originate from postgerminal center B cells and share a remarkable alteration in B cell transcription factor expression and/or activation with classical Hodgkin's disease cells. Comparative analysis of gene expression by cDNA microarray of BCBL-1 cells (PEL), L-428 (classical Hodgkin's disease), and BJAB cells revealed a subset of genes that were differentially expressed in BCBL 1 cells. Among these, four genes involved in cell migration and chemotaxis were strongly up-regulated in PEL cells: leukotriene A4 (LTA4) hydrolase (LTA4H), IL 16, thrombospondin-1 (TSP-1), and selectin-P ligand (PSGL-1). Up-regulation of LTA4H was investigated at the transcriptional level. Full-length LTA4H promoter exhibited 50% higher activity in BCBL-1 cells than in BJAB or L-428 cells. Deletion analysis of the LTA4H promoter revealed a positive cis-regulatory element active only in BCBL-1 cells in the promoter proximal region located between -76 and -40 bp. Formation of a specific DNA-protein complex in this region was confirmed by EMSA. Coculture of ionophore-stimulated primary neutrophils with BCBL-1 cells leads to an increased production of LTB4 compared with coculture with BJAB and L-428 cells as measured by enzyme immunoassay, demonstrating the functional significance of LTA4H up-regulation. PMID- 16709868 TI - The histamine H4 receptor mediates allergic airway inflammation by regulating the activation of CD4+ T cells. AB - Histamine is an important inflammatory mediator that is released in airways during an asthmatic response. However, current antihistamine drugs are not effective in controlling the disease. The discovery of the histamine H4 receptor (H4R) prompted us to reinvestigate the role of histamine in pulmonary allergic responses. H4R-deficient mice and mice treated with H4R antagonists exhibited decreased allergic lung inflammation, with decreases in infiltrating lung eosinophils and lymphocytes and decreases in Th2 responses. Ex vivo restimulation of T cells showed decreases in IL-4, IL-5, IL-13, IL-6, and IL-17 levels, suggesting that T cell functions were disrupted. In vitro studies indicated that blockade of the H4R on dendritic cells leads to decreases in cytokine and chemokine production and limits their ability to induce Th2 responses in T cells. This work suggests that the H4R can modulate allergic responses via its influence on T cell activation. The study expands the known influences of histamine on the immune system and highlights the therapeutic potential of H4R antagonists in allergic conditions. PMID- 16709869 TI - Diagnostic and therapeutic potential of amyloid-reactive IgG antibodies contained in human sera. AB - Passive immunotherapy using fibril-reactive mAbs has been shown experimentally to reduce amyloid formation and also accelerate amyloidolysis. We now report that human sera, as well as various sources of pooled human IgG, including pharmacologic formulations of immune globulin i.v. (IGIV), contain Abs that specifically recognize fibrils formed from light chains and other amyloidogenic precursor proteins, including serum amyloid A, transthyretin, islet amyloid polypeptide, and amyloid beta 1-40 peptide, but notably, do not react with these molecules in their native nonfibrillar forms. After isolation of the Abs from IGIV via fibril-conjugated affinity column chromatography, the EC50-binding value for light chains and amyloid beta 1-40 peptide fibrils was approximately 15 nM-a magnitude approximately 200 and 70 times less than that of the unbound fraction and unfractionated product, respectively. Comparable reactivity was found in the case of those formed from serum amyloid A, transthyretin, and islet amyloid polypeptide. The purified Abs immunostained human amyloid tissue deposits and could inhibit fibrillogenesis, as shown in fibril formation and extension assays. Most importantly, in vivo reactivity was evidenced in a murine model when the enriched Abs were used to image amyloid, as well as expedite its removal. These promising experimental results suggest that fibril affinity-purified IGIV has potential as a diagnostic and therapeutic agent for patients with amyloid associated disease. PMID- 16709870 TI - The role of noradrenergic nerves in the development of the lymphoproliferative disease in Fas-deficient, lpr/lpr mice. AB - Lpr/lpr mice develop a lymphoproliferative, autoimmune, lupus-like disease. These mice lack functional Fas (CD95) expression and are resistant to Fas ligand (CD178)-mediated apoptosis, a critical mechanism for the maintenance of peripheral tolerance. In this study, we show that noradrenaline (NA), the main sympathetic neurotransmitter, can induce apoptosis of lymphoid cells independently of functional Fas. Based on this finding, we used lpr/lpr mice as model to study the role of noradrenergic nerves in the expression of a lymphoproliferative disease. Early in ontogeny, the concentration of NA was significantly increased in the spleen of lpr/lpr mice, compared with normal littermates. However, splenic sympathetic innervation gradually declined as the disease progressed, and IgM blood levels and splenic NA concentration inversely correlated when the disease was overtly manifested. When the loss of noradrenergic fibers that occurred naturally during adult life in lpr/lpr mice was experimentally advanced by neonatal sympathectomy, the concentration of IgM and IgG2a in blood was markedly higher than that of control lpr/lpr mice, and the appearance of lymphadenopathy was accelerated. Furthermore, although neonatal denervation did not affect the life span of normal animals, it shortened significantly the survival time of lpr/lpr mice. These data show that, in addition to defects in the Fas pathway, an altered sympathetic innervation in lpr/lpr mice also contributes to the pathogenesis of the autoimmune disease, and strongly support the hypothesis that the sympathetic nervous system can modulate the expression of lymphoproliferative diseases. PMID- 16709871 TI - CXCR3 and its ligands in a murine model of obliterative bronchiolitis: regulation and function. AB - Lung transplantation remains the only effective therapy for patients with end stage lung disease, but survival is limited by the development of obliterative bronchiolitis (OB). The chemokine receptor CXCR3 and two of its ligands, CXCL9 and CXCL10, have been identified as important mediators of OB. However, the relative contribution of CXCL9 and CXCL10 to the development of OB and the mechanism of regulation of these chemokines has not been well defined. In this study, we demonstrate that CXCL9 and CXCL10 are up-regulated in unique patterns following tracheal transplantation in mice. In these experiments, CXCL9 expression peaked 7 days posttransplant, while CXCL10 expression peaked at 1 day and then again 7 days posttransplant. Expression of CXCL10 was also up-regulated in a novel murine model of lung ischemia, and in bronchoalveolar lavage fluid taken from human lungs 24 h after lung transplantation. In further analysis, we found that 3 h after transplantation CXCL10 is donor tissue derived and not dependent on IFN-gamma or STAT1, while 24 h after transplantation CXCL10 is from recipient tissue and regulated by IFN-gamma and STAT1. Expression of both CXCL9 and CXCL10 7 days posttransplant is regulated by IFN-gamma and STAT1. Finally, we demonstrate that deletion of CXCR3 in recipients reduces airway obliteration. However, deletion of either CXCL9 or CXCL10 did not affect airway obliteration. These data show that in this murine model of obliterative bronchiolitis, these chemokines are differentially regulated following transplantation, and that deletion of either chemokine alone does not affect the development of airway obliteration. PMID- 16709872 TI - Patients with chronic granulomatous disease have a reduced peripheral blood memory B cell compartment. AB - In this study, we have identified an altered B cell compartment in patients with chronic granulomatous disease (CGD), a disorder of phagocyte function, characterized by pyogenic infections and granuloma formation caused by defects in NADPH activity. This is characterized by an expansion of CD5-expressing B cells, and profound reduction in B cells expressing the memory B cell marker, CD27. Both findings were independent of the age, genotype, and clinical status of the patients, and were not accompanied by altered CD5 and CD27 expression on T cells. Focusing on CD27-positive B cells, considered to be memory cells based on somatically mutated Ig genes, we found that the reduction was not caused by CD27 shedding or abnormal retention of CD27 protein inside the cell. Rather, it was determined that CD27-negative B cells were, appropriately, CD27 mRNA negative, consistent with a naive phenotype, whereas CD27-positive B cells contained abundant CD27 mRNA and displayed somatic mutations, consistent with a memory B cell phenotype. Thus, it appears that CGD is associated with a significant reduction in the peripheral blood memory B cell compartment, but that the basic processes of somatic mutation and expression of CD27 are intact. X-linked carriers of CGD revealed a significant correlation between the percentage of CD27 positive B cells and the percentage of neutrophils with normal NADPH activity, reflective of the degree of X chromosome lyonization. These results suggest a role for NADPH in the process of memory B cell formation, inviting further exploration of secondary Ab responses in CGD patients. PMID- 16709873 TI - Identical TCR beta-chain rearrangements in streptococcal angina and skin lesions of patients with psoriasis vulgaris. AB - Tonsillar infection with Streptococcus pyogenes may induce several nonsuppurative autoimmune sequelae. The precise pathogenetic mechanisms behind this clinically well-established association are still unresolved. Using TCR analysis, we sought to identify a link between streptococcal tonsillitis and the T cell-mediated autoimmune response in psoriasis. Three patients with streptococcal-induced psoriasis underwent tonsillectomy. Using size spectratyping and sequencing of TCR beta-chain variable region gene (TCRBV) rearrangements, we compared the TCR usage of psoriatic skin lesions, blood, tonsils, and tonsillar T cells fractionated according to the expression of the skin address in "cutaneous lymphocyte associated Ag" (CLA). TCRBV-size spectratype analysis of the blood lymphocytes, tonsils, and the CLA-negative tonsillar T cells revealed largely unselected T cell populations. Instead, TCRBV gene families of the psoriatic lesions and skin homing CLA-positive tonsillar T cells displayed highly restricted spectratypes. Sequencing of TCRBV cDNA identified various clonal TCRBV rearrangements within the psoriatic lesions that indicated Ag-driven T cell expansion. Several of these clonotypes were also detected within the tonsils and, in one of the patients, within the small subset of CLA-positive tonsillar T cells, suggesting that T cells from the same T cell clones were simultaneously present within skin and tonsillar tissue. Because after tonsillectomy psoriasis cleared in all three patients our observations indicate that T cells may connect psoriatic inflammation to streptococcal angina. They suggest that the chronic streptococcal immune stimulus within the tonsils could act as a source for pathogenic T cells in poststreptococcal disorders, and they may help to explain why eliminating this source with tonsillectomy may improve streptococcal-induced sequelae. PMID- 16709874 TI - Carriage of DRB1*13 is associated with increased posttreatment IgE levels against Schistosoma mansoni antigens and lower long-term reinfection levels. AB - Praziquantel treatment for Schistosoma mansoni infection enhances Th2 responsiveness against parasite Ags, but also increases the variance in Ab isotype levels. This effect may arise partly from genetic heterogeneity. In this study, associations between HLA polymorphisms at three loci (HLA-DQB1, HLA-DQA1, and HLA-DRB1) and posttreatment Ig responses to S. mansoni Ags were assessed in 199 individuals aged 7-50 years from Uganda. Blood samples were assayed for IgG1, IgG4, and IgE levels against soluble worm Ag (SWA), soluble egg Ag, tegument Ag, and a recombinant tegumental Ag (rSm 22.6) 7 wk after treatment. Multivariate ANOVA analysis initially revealed associations between carriage of DRB1*13 and increased levels of IgG1, IgG4, and IgE against SWA, tegument Ag, and rSM22.6. Subsequent analysis of covariance, which controlled for correlations between isotype levels and also included pretreatment IL-4, IL-5, and IL-13 responsiveness against SWA as covariates, revealed an independent association only between DRB1*13 and a factor score summarizing IgE levels to worm-derived Ags, which was strongest in adults. A post hoc age- and sex-stratified analysis revealed lower reinfection intensities at 1 year, 22 mo, and 6 years after the first round of treatment among carriers of DRB1*13. These results indicate that genetic background has a prominent influence on the posttreatment Th2 immune response to S. mansoni Ags, as well as a downstream association with long-term reinfection levels. PMID- 16709875 TI - Therapeutic induction of regulatory, cytotoxic CD8+ T cells in multiple sclerosis. AB - In the setting of autoimmunity, one of the goals of successful therapeutic immune modulation is the induction of peripheral tolerance, a large part of which is mediated by regulatory/suppressor T cells. In this report, we demonstrate a novel immunomodulatory mechanism by an FDA-approved, exogenous peptide-based therapy that incites an HLA class I-restricted, cytotoxic suppressor CD8+ T cell response. We have shown previously that treatment of multiple sclerosis (MS) with glatiramer acetate (GA; Copaxone) induces differential up-regulation of GA reactive CD8+ T cell responses. We now show that these GA-induced CD8+ T cells are regulatory/suppressor in nature. Untreated patients show overall deficit in CD8+ T cell-mediated suppression, compared with healthy subjects. GA therapy significantly enhances this suppressive ability, which is mediated by cell contact-dependent mechanisms. CD8+ T cells from GA-treated patients and healthy subjects, but not those from untreated patients with MS, exhibit potent, HLA class I-restricted, GA-specific cytotoxicity. We further show that these GA induced cytotoxic CD8+ T cells can directly kill CD4+ T cells in a GA-specific manner. Killing is enhanced by preactivation of target CD4+ T cells and may depend on presentation of GA through HLA-E. Thus, we demonstrate that GA therapy induces a suppressor/cytotoxic CD8+ T cell response, which is capable of modulating in vivo immune responses during ongoing therapy. These studies not only explain several prior observations relating to the mechanism of this drug but also provide important insights into the natural immune interplay underlying this human immune-mediated disease. PMID- 16709877 TI - Treatment and transfer of emphysema by a new bone marrow transplantation method from normal mice to Tsk mice and vice versa. AB - We have recently established a new bone marrow transplantation (BMT) method in which bone marrow cells are injected into the intrabone marrow (IBM). In the present study, we used an animal model for emphysema (tight-skin [Tsk] mouse) to examine whether IBM-BMT could be used to treat emphysema in Tsk mice. IBM-BMT was carried out from C3H mice into Tsk mice (8-10 weeks old) that had already shown emphysema. Six months after transplantation, the lungs of all the Tsk mice treated with IBM-BMT [C3H-->Tsk] showed similar structures to those of normal mice, whereas the [Tsk-->Tsk] mice showed emphysema, as seen in age-matched Tsk mice. Next, we attempted to transfer emphysema from Tsk mice to C3H mice by IBM BMT. Six months after IBM-BMT, the [Tsk-->C3H] mice showed emphysema. These results strongly suggest that emphysema in Tsk mice originates from defects of stem cells in the bone marrow. PMID- 16709878 TI - Confronting infectious diseases is a perpetual struggle, top official says. PMID- 16709880 TI - USP chapter 797 enforceable but not often enforced. PMID- 16709876 TI - Reprogramming efficiency following somatic cell nuclear transfer is influenced by the differentiation and methylation state of the donor nucleus. AB - Reprogramming of a differentiated cell nucleus by somatic cell nuclear transplantation is an inefficient process. Following nuclear transfer, the donor nucleus often fails to express early embryonic genes and establish a normal embryonic pattern of chromatin modifications. These defects correlate with the low number of cloned embryos able to produce embryonic stem cells or develop into adult animals. Here, we show that the differentiation and methylation state of the donor cell influence the efficiency of genomic reprogramming. First, neural stem cells, when used as donors for nuclear transplantation, produce embryonic stem cells at a higher efficiency than blastocysts derived from terminally differentiated neuronal donor cells, demonstrating a correlation between the state of differentiation and cloning efficiency. Second, using a hypomorphic allele of DNA methyltransferase-1, we found that global hypomethylation of a differentiated cell genome improved cloning efficiency. Our results provide functional evidence that the differentiation and epigenetic state of the donor nucleus influences reprogramming efficiency. PMID- 16709881 TI - Hospitals must improve infrastructure, surge capacity, officials say. PMID- 16709882 TI - CMS targets pharmacy quality. PMID- 16709884 TI - Pharmacists' attitudes toward ACLS provider training program. PMID- 16709883 TI - Enhancing quality and safety through physician-pharmacist collaboration. PMID- 16709885 TI - Pharmacist Current Procedural Terminology codes and medication therapy management. PMID- 16709886 TI - New ASHP pharmacy residency accreditation standards. PMID- 16709887 TI - Using preprinted medication order forms to improve the safety of investigational drug use. PMID- 16709888 TI - Experiential rotations for pharmacy students. PMID- 16709889 TI - Reflections on international pharmacy. PMID- 16709890 TI - Drug therapy during labor and delivery, part 1. AB - PURPOSE: The drug therapy of common conditions and complications during labor and delivery and the fetal and neonatal effects of this therapy are examined. SUMMARY: The pharmacologic therapy of common conditions that occur in labor and delivery primarily involves oxytocin and prostaglandins for cervical ripening and labor induction and systemic and regional narcotic analgesics for pain. Because most medications used in women during labor and delivery do not have Food and Drug Administration-approved labeling, pharmacists should understand the benefits and limitations of medications used in the mother. Although induction and augmentation of labor and the control of pain often require drug therapy, other, less frequent, complications may occur in labor. Drug therapies for these complications include anti-infective agents to treat maternal infection and prevent neonatal diseases; antiretrovirals to reduce perinatal HIV-1 transmission from the mother to the fetus; corticosteroids to prevent fetal lung immaturity; antihypertensives to treat preeclampsia; anticonvulsants to treat eclampsia; antibiotics to prolong pregnancy and improve neonatal outcomes after premature rupture of the membranes; tocolytics for premature labor; and oxytocin, ergot alkaloids, and prostaglandin analogues for postpartum hemorrhage. The fetal and neonatal effects of therapy for the conditions that occur during labor and delivery are usually benign, but significant morbidity and mortality involving the mother, the fetus, and the newborn are ever-present risks. CONCLUSION: Awareness of the conditions and complications requiring drug therapy during labor and delivery will allow hospital pharmacists to make knowledgeable decisions about the rapid accessibility of critical medications in the labor and delivery unit. PMID- 16709892 TI - Geometric probability distribution for modeling of error risk during prescription dispensing. AB - PURPOSE: The relationship between the number of prescriptions dispensed by individual pharmacy staff during a single workday and the probability of committing at least one dispensing error during that same workday period was evaluated using a geometric probability distribution. SUMMARY: A cross-sectional descriptive study involving 50 pharmacies located in six cities across the United States was conducted. A pharmacist trained to detect dispensing errors recorded the number of prescriptions filled by each pharmacy staff member and noted which prescription represented the staff member's first dispensing error (defined as any deviation from the prescriber's order) made during the observation period. The Kolmogorov-Smirnov tests for discrete distributions revealed that the observed cumulative distribution of dispensing errors could have come from a geometric probability distribution that assumed dispensing error rates of 2-3%. In terms of risk analysis, this study's findings suggest that there can be a quantifiable statistical relationship between a measure of workload and the risk of committing at least one dispensing error. The ability to model dispensing errors using a geometric probability distribution enables the safety and health care practitioner to directly assess dispensing error risk as a function of a pharmacy's accuracy rate and the number of prescriptions a pharmacy staff member should dispense during a work shift. CONCLUSION: A geometric probability distribution effectively modeled the relationship between the number of prescriptions filled and the occurrence of the first dispensing errors. PMID- 16709891 TI - Misunderstanding of prescription drug warning labels among patients with low literacy. AB - PURPOSE: The common causes for misunderstanding prescription drug warning labels (PWLs) among adults with low literacy were studied. METHODS: A total of 74 patients reading at or below the sixth-grade level and receiving care at the primary care clinic at the Louisiana State University Health Sciences Center in Shreveport were recruited to participate in structured interviews. Patients were asked to interpret and comment on eight commonly used warning labels found on prescription medications. Correct interpretation was determined by expert panel review of patients' verbatim responses. Qualitative methods were employed to code responses and generate themes regarding the misunderstanding of these PWLs. RESULTS: Among this sample of patients with low literacy skills, rates of correct interpretation for the eight warning labels ranged from 0% to 78.7%. With the exception of the most basic label, less than half of all patients were able to provide adequate interpretations of the warning label messages. Five themes were derived to describe the common causes for misunderstanding the labels: single step versus multiple-step instructions, reading difficulty of text, use of icons, use of color, and message clarity. Labels were at greater risk for being misunderstood if they included multiple instructions, had a greater reading difficulty, included unfamiliar terms, or used confusing icons that were discordant with text messages. Participants also frequently imposed an incorrect meaning on label colors, which led to further confusion. CONCLUSION: Patients with low literacy skills demonstrated a lower rate of correct interpretation of the eight most commonly used PWLs than did those with higher literacy skills. Multiple-step instructions, reading difficulty of text, the use of icons, the use of color, and message clarity were the common causes of label misinterpretation. PMID- 16709893 TI - Evaluating the off-label use of medications in children. PMID- 16709894 TI - Therapeutic conversion from fosinopril to benazepril at a Veterans Affairs medical center. PMID- 16709895 TI - Affirmation of states' authority to define "legitimate medical purpose". PMID- 16709896 TI - ASHP Therapeutic Position Statement on the Treatment of Hypertension. PMID- 16709897 TI - Selectivity of connexin 43 channels is regulated through protein kinase C dependent phosphorylation. AB - Coordinated contractile activation of the heart and resistance to ischemic injury depend, in part, on the intercellular communication mediated by Cx43-composed gap junctions. The function of these junctions is regulated at multiple levels (assembly to degradation) through phosphorylation at specific sites in the carboxyl terminus (CT) of the Cx43 protein. We show here that the selective permeability of Cx43 junctions is regulated through protein kinase C (PKC) dependent phosphorylation at serine 368 (S368). Selective permeability was measured in several Cx43-expressing cell lines as the rate constant for intercellular dye diffusion relative to junctional conductance. The selective permeability of Cx43 junctions under control conditions was quite variable, as was the open-state behavior of the comprising channels. Coexpression of the CT of Cx43 as a distinct protein, treatment with a PKC inhibitor, or mutation of S368 to alanine, all reduced (or eliminated) phosphorylation at S368, reduced the incidence of 55- to 70-pS channels, and reduced by 10-fold the selective permeability of the junctions for a small cationic dye. Because PKC activation during preischemic conditioning is cardioprotective during subsequent ischemic episodes, we examined no-flow, ischemic hearts for Cx43 phosphorylated at S368 (pS368). Consistent with early activation of PKC, pS368-Cx43 was increased in ischemic hearts; despite extensive lateralization of total Cx43, pS368-Cx43 remained predominantly at intercalated disks. Our data suggest that the selectivity of gap junction channels at intercalated disks is increased early in ischemia. PMID- 16709898 TI - Formation of the venous pole of the heart from an Nkx2-5-negative precursor population requires Tbx18. AB - The venous pole of the mammalian heart is a structurally and electrically complex region, yet the lineage and molecular mechanisms underlying its formation have remained largely unexplored. In contrast to classical studies that attribute the origin of the myocardial sinus horns to the embryonic venous pole, we find that the sinus horns form only after heart looping by differentiation of mesenchymal cells of the septum transversum region into myocardium. The myocardial sinus horns and their mesenchymal precursor cells never express Nkx2-5, a transcription factor critical for heart development. In addition, lineage studies show that the sinus horns do not derive from cells previously positive for Nkx2-5. In contrast, the sinus horns express the T-box transcription factor gene Tbx18. Mice deficient for Tbx18 fail to form sinus horns from the pericardial mesenchyme and have defective caval veins, whereas the pulmonary vein and atrial structures are unaffected. Our studies define a novel heart precursor population that contributes exclusively to the myocardium surrounding the sinus horns or systemic venous tributaries of the developing heart, which are a source of congenital malformation and cardiac arrhythmias. PMID- 16709899 TI - Hypoxia inducible factor 1 mediates hypoxia-induced TRPC expression and elevated intracellular Ca2+ in pulmonary arterial smooth muscle cells. AB - Chronic hypoxia (CH) causes pulmonary vasoconstriction because of increased pulmonary arterial smooth muscle cell (PASMC) contraction and proliferation. We previously demonstrated that intracellular Ca(2+) concentration ([Ca(2+)](i)) was elevated in PASMCs from chronically hypoxic rats because of Ca(2+) influx through pathways other than L-type Ca(2+) channels and that development of hypoxic pulmonary hypertension required full expression of the transcription factor hypoxia inducible factor 1 (HIF-1). In this study, we examined the effect of CH on the activity and expression of store-operated Ca(2+) channels (SOCCs) and the regulation of these channels by HIF-1. Capacitative Ca(2+) entry (CCE) was enhanced in PASMCs from intrapulmonary arteries of rats exposed to CH (10% O(2); 21 days), and exposure to Ca(2+)-free extracellular solution or SOCC antagonists (SKF96365 or NiCl(2)) decreased resting [Ca(2+)](i) in these cells. Expression of TRPC1 and TRPC6, but not TRPC4, mRNA and protein was increased in PASMCs from rats and wild-type mice exposed to CH, in PASMCs from normoxic animals cultured under hypoxic conditions (4% O(2); 60 hours), and in PASMCs in which HIF-1 was overexpressed under nonhypoxic conditions. Hypoxia-induced increases in basal [Ca(2+)](i) and TRPC expression were absent in mice partially deficient for HIF 1. These results suggest that increased TRPC expression, leading to enhanced CCE through SOCCs, may contribute to hypoxic pulmonary hypertension by facilitating Ca(2+) influx and increasing basal [Ca(2+)](i) in PASMCs and that this response is mediated by HIF-1. PMID- 16709900 TI - An osteopontin-NADPH oxidase signaling cascade promotes pro-matrix metalloproteinase 9 activation in aortic mesenchymal cells. AB - Osteopontin (OPN) is a cytokine upregulated in diabetic vascular disease. To better understand its role in vascular remodeling, we assessed how OPN controls metalloproteinase (MMP) activation in aortic adventitial myofibroblasts (AMFs) and A7r5 vascular smooth muscle cells (VSMCs). By zymography, OPN and tumor necrosis factor (TNF)-alpha preferentially upregulate pro-matrix metalloproteinase 9 (pro-MMP9) activity. TNF-alpha upregulated pro-MMP9 in AMFs isolated from wild-type (OPN(+/+)) mice, but pro-MMP9 induction was abrogated in AMFs from OPN(-/-) mice. OPN treatment of VSMCs enhanced pro-MMP9 activity, and TNF-alpha induction of pro-MMP9 was inhibited by anti-OPN antibody and apocynin. Superoxide and the oxylipid product 8-isoprostaglandin F(2) alpha-isoprostane (8 IsoP) were increased by OPN treatment, and anti-OPN antibody suppressed 8-IsoP production. Like OPN and TNF-alpha, 8-IsoP preferentially activated pro-MMP9. Superoxide, 8-IsoP, and NADPH oxidase 2 (Nox2) subunits were reduced in OPN(-/-) AMFs. Treatment of A7r5 VSMCs with OPN upregulated NADPH oxidase subunit accumulation. OPN structure/function studies mapped these activities to the SVVYGLR heptapeptide motif in the thrombin-liberated human OPN N-terminal domain (SLAYGLR in mouse OPN). Treatment of aortic VSMCs with SVVYGLR upregulated pro MMP9 activity and restored TNF-alpha activation of pro-MMP9 in OPN(-/-) AMFs. Injection of OPN-deficient OPN(+/-) mice with SVVYGLR peptide upregulated pro MMP9 activity, 8-IsoP levels, and Nox2 protein levels in aorta and increased panmural superoxide production (dihydroethidium staining). At equivalent hyperglycemia and dyslipidemia, 8-IsoP levels and aortic pro-MMP9 were reduced with complete OPN deficiency in a model of diet-induced diabetes, achieved by comparing OPN(-/-)/LDLR(-/-) versus OPN(+/-)/LDLR(-/-) siblings. Thus, OPN provides a paracrine signal that augments vascular pro-MMP9 activity, mediated in part via superoxide generation and oxylipid formation. PMID- 16709901 TI - Maximum phosphorylation of the cardiac ryanodine receptor at serine-2809 by protein kinase a produces unique modifications to channel gating and conductance not observed at lower levels of phosphorylation. AB - It is suggested that protein kinase A (PKA)-dependent phosphorylation of cardiac ryanodine receptors (RyR2) is linked to the development of heart failure and the generation of fatal cardiac arrhythmias. It is also suggested that RyR2 is phosphorylated to 75% of maximum levels in heart failure resulting in leaky, unregulated channels gating in subconductance states. We now demonstrate that this is unlikely, as RyR2 isolated from nonfailing cardiac muscle is phosphorylated to 75% of maximum at serine-2809, and in this situation, RyR2 displays low open probability (P(o)) (0.059+/-0.010 [SEM]; n=30) and normal regulation of gating by Ca(2+) and other ligands. However, when serine-2809 is PKA phosphorylated to maximum levels on RyR2, unique changes in channel behavior are observed. The channel displays enhanced single-channel conductance, very long open states causing large increases in P(o), and no evidence of subconductance states. Dephosphorylation of channels by protein phosphatase 1 (from 75% to near 0% at serine-2809) also enhances RyR2 channel activity through abbreviation of closed lifetimes. We propose that channels phosphorylated to 75% of maximum at serine-2809 occupy a natural low point in the RyR2 activity landscape. This optimizes channel control, which can be accomplished either by enhanced or decreased phosphorylation, making the channel particularly sensitive to the kinase:phosphatase balance. Pathological situations such as heart failure might upset this balance and thereby permit prolonged stoichiometric phosphorylation of serine-2809, which would be required for dysregulation of SR Ca(2+) release. PMID- 16709902 TI - Neural crest cells retain multipotential characteristics in the developing valves and label the cardiac conduction system. AB - Multipotent neural crest cells (NCCs) are a major extracardiac component of cardiovascular development. Although recognized as contributing cells to the arterial valves at early developmental stages, NCC persistence in the valves at later times or in the adult heart is controversial. We analyzed NCC persistence and contributions to both semilunar and atrioventricular (AV) valves in the mature heart. Two NCC-specific promoters driving Cre recombinase, Wnt1-Cre and P0 Cre, were mated with floxed reporter mice, R26R or CAG-CAT-EGFP, to map NCC fate. Hearts were analyzed before aorticopulmonary (AP) septation through adult stages. As previously demonstrated, strong NCC labeling was detected in ventral and dorsal outflow cushions before AP septation. In contrast to previous reports, we found that substantial numbers of labeled cells persisted in the semilunar valves in late fetal, neonatal, and adult hearts. Furthermore, NCCs were also found in the AV valves, almost exclusively in the septal leaflets. NCCs in the AV valves expressed melanocytic and neurogenic markers. However, cells labeled in the proximal cardiac conduction system exhibited neurogenic and gliagenic markers, whereas some NCCs expressed no differentiation specific markers. These results suggest that cardiac NCCs contribute to the mature valves and the cardiac conduction system and retain multipotent characteristics late in development. PMID- 16709903 TI - Deciphering the organization and modulation of spinal locomotor central pattern generators. AB - Networks within our spinal cord generate the basic pattern underlying walking. Over the past decade, much progress has been made in our understanding of their function in a variety of vertebrate species. A significant hurdle has been the identification of candidate populations of neurons that are involved in pattern generation in the spinal cord. Recently, systems neuroscientists in collaboration with molecular biologists have begun to dissect the circuitry underlying spinal locomotor networks. These advances have combined genetic and electrophysiological techniques using in vitro preparations of the mouse spinal cord. This review will discuss new advances in the field of spinal locomotor networks with emphasis on the mouse. Many of the behaviors fundamental to animal life, such as breathing, chewing and locomotion, are rhythmic activities controlled by neuronal networks. Discerning which neurons are members of these networks, their synaptic connectivity and their individual electrophysiological properties is essential to our understanding of how rhythmic motor behaviors are produced. It is well known that the spinal cord contains the basic circuitry to produce locomotion. However, identifying neurons and connections within spinal networks is challenging because cells that comprise the locomotor network form part of a heterogeneous mix of interneurons within the ventral spinal cord. Recently, the merging of electrophysiological and genetic approaches has provided new tools to identify classes of interneurons within the spinal cord that contribute to network function. These new findings will be discussed in this review. PMID- 16709904 TI - Activation of a sensorimotor pathway in response to a water temperature drop in a teleost fish. AB - When common carp, Cyprinus carpio L., experience a rapid temperature drop, the cerebral blood volume is strongly reduced to dampen the temperature drop in the brain. Simultaneously, the preoptic area and pituitary gland are activated to launch whole-body adaptive responses. However, the preferred reaction of fish to a temperature change is an escape reaction, which implies activation of a sensorimotor pathway. Here, we used blood oxygenation level-dependent (BOLD)- and cerebral blood volume (CBV)-weighted functional magnetic resonance imaging (fMRI) to identify a sensorimotor pathway, during a 10 degrees C temperature drop in common carp. Transient activation was observed in the region where the sensory root of the trigeminal nerve enters the brain, and in the valvula cerebelli. In both regions, metabolic activity increased (increased deoxyhemoglobin content demonstrated by a decreased BOLD signal) within 30 s after the onset of the temperature drop, peaked after 2-3 min, and then decreased, even though the temperature continued to drop for another 2 min. These brain structures appear to respond to temperature change, rather than to the absolute temperature. Thus, during a temperature drop, the sensorimotor pathway consisting of the trigeminal nerve, the primary sensory trigeminal nucleus, the valvula cerebelli and some motornuclei, is active, in line with perception of temperature change in the buccal cavity, leading to motor activity for escape. This pathway operates in parallel to an acclimation pathway, which involves the preoptic area to pituitary gland pathway. PMID- 16709905 TI - Fast-swimming hydromedusae exploit velar kinematics to form an optimal vortex wake. AB - Fast-swimming hydromedusan jellyfish possess a characteristic funnel-shaped velum at the exit of their oral cavity that interacts with the pulsed jets of water ejected during swimming motions. It has been previously assumed that the velum primarily serves to augment swimming thrust by constricting the ejected flow in order to produce higher jet velocities. This paper presents high-speed video and dye-flow visualizations of free-swimming Nemopsis bachei hydromedusae, which instead indicate that the time-dependent velar kinematics observed during the swimming cycle primarily serve to optimize vortices formed by the ejected water rather than to affect the speed of the ejected flow. Optimal vortex formation is favorable in fast-swimming jellyfish because, unlike the jet funnelling mechanism, it allows for the minimization of energy costs while maximizing thrust forces. However, the vortex ;formation number' corresponding to optimality in N. bachei is substantially greater than the value of 4 found in previous engineering studies of pulsed jets from rigid tubes. The increased optimal vortex formation number is attributable to the transient velar kinematics exhibited by the animals. A recently developed model for instantaneous forces generated during swimming motions is implemented to demonstrate that transient velar kinematics are required in order to achieve the measured swimming trajectories. The presence of velar structures in fast-swimming jellyfish and the occurrence of similar jet regulating mechanisms in other jet-propelled swimmers (e.g. the funnel of squid) appear to be a primary factor contributing to success of fast-swimming jetters, despite their primitive body plans. PMID- 16709906 TI - Spectral sensitivity of the two-spotted goby Gobiusculus flavescens (Fabricius): a physiological and behavioural study. AB - Microspectrophotometry of Gobiusculus flavescens photoreceptors revealed a single rod visual pigment (lambdamaxat 508 nm) and the three cone pigments (lambdamax 456, 531 and 553 nm). The cone population was dominated by identical double cones containing the middle-wave-sensitive (MWS) pigment, but with a small number of non-identical MWS/LWS (long-wave-sensitive) and identical LWS double cones. Small populations of large single cones also contained either the MWS or LWS pigment. The short-wave-sensitive (SWS) pigment was found in small single cones. Lens transmission was great reduced below 410 nm. The spectral sensitivity of the behaviourally determined reaction distance (RD) to prey at a high irradiance level (0.5 micromol m-2 s-1) correlated with the maximum sensitivity of the MWS cones, both peaking around 530 nm. However, at a lower irradiance level (0.015 micromol m-2 s-1) such a correlation was not so apparent. The RD was greatly reduced, though still maintaining a peak around 530-550 nm, but with a relatively smaller reduction in RD at shorter wavelengths. Optomotor behaviour displayed a somewhat similar spectral sensitivity to the RD responses at the higher light intensity. However, the peak was at slightly longer wavelengths at 550 nm, suggesting a greater input from LWS cones to the optomotor response. PMID- 16709907 TI - Adaptive value of ambling gaits in primates and other mammals. AB - At speeds between the walk and the gallop, most mammals trot. Primates almost never trot, and it has been claimed that they transition directly from a walk to a gallop without any distinctive mid-speed running gait. If true, this would be another characteristic difference between the locomotion of primates and that of most other quadrupedal mammals. Presently, however, few data exist concerning the actual presence or absence of intermediate-speed gaits (i.e. gaits that are used between a walk and a gallop) in primates. Video records of running in twelve primate species reveal that, unlike most other mammals, all the primates studied almost exclusively adopt an 'amble'--an intermediate-speed running gait with no whole-body aerial phase--rather than trot. Ambling is also common in elephants and some horses, raising the question of why ambling is preferred over trotting in these diverse groups of animals. Mathematical analyses presented here show that ambling ensures continuous contact of the body with the substrate while dramatically reducing vertical oscillations of the center of mass. This may explain why ambling appears to be preferable to trotting for extremely large terrestrial mammals such as elephants and for arboreal mammals like primates that move on unstable branches. These findings allow us to better understand the mechanics of these unusual running gaits and shed new light on primate locomotor evolution. PMID- 16709908 TI - The energetic costs of trunk and distal-limb loading during walking and running in guinea fowl Numida meleagris: I. Organismal metabolism and biomechanics. AB - We examined the energetic cost of loading the trunk or distal portion of the leg in walking and running guinea fowl (Numida meleagris). These different loading regimes were designed to separately influence the energy use by muscles used during the stance and swing phases of the stride. Metabolic rate, estimated from oxygen consumption, was measured while birds locomoted on a motorized treadmill at speeds from 0.5 to 2.0 m s-1, either unloaded, or with a mass equivalent to 23% of their body mass carried on their backs, or with masses equal to approximately 2.5% of their body mass attached to each tarsometatarsal segment. In separate experiments, we also measured the duration of stance and swing in unloaded, trunk-loaded, or limb-loaded birds. In the unloaded and limb-loaded birds, we also calculated the mechanical energy of the tarsometatarsal segment throughout the stride. Trunk and limb loads caused similar increases in metabolic rate. During trunk loading, the net metabolic rate (gross metabolic rate-resting metabolic rate) increased by 17% above the unloaded value across all speeds. This percentage increase is less than has been found in most studies of humans and other mammals. The economical load carriage of guinea fowl is consistent with predictions based on the relative cost of the stance and swing phases of the stride in this species. However, the available comparative data and considerations of the factors that determine the cost of carrying extra mass lead us to the conclusion that the cost of load carrying is unlikely to be a reliable indicator of the distribution of energy use in stance and swing. Both loading regimes caused small changes in the swing and/or stance durations, but these changes were less than 10%. Loading the tarsometatarsal segment increased its segmental energy by 4.1 times and the segmental mechanical power averaged over the stride by 3.8 times. The increases in metabolism associated with limb loading appear to be linked to the increases in mechanical power. The delta efficiency (change in mechanical power divided by the change in metabolic power) of producing this power increased from 11% in walking to approximately 25% in running. Although tarsometatarsal loading was designed to increase the mechanical energy during swing phase, 40% of the increase in segmental energy occurred during late stance. Thus, the increased energy demand of distal limb loading in guinea fowl is predicted to cause increases in energy use by both stance- and swing-phase muscles. PMID- 16709909 TI - The energetic costs of trunk and distal-limb loading during walking and running in guinea fowl Numida meleagris: II. Muscle energy use as indicated by blood flow. AB - We examined the changes in muscle energy use in guinea fowl running at 1.5 m s-1 either unloaded, or carrying trunk loads equal to 23% of body mass, or loads on their distal legs equal to a total of 5% of body mass. We estimated muscle energy use by measuring blood flow to all of the leg muscles using injected microspheres. Total blood flow to the leg muscles increased by approximately 15% under both loading conditions, which matched the percentage increase in net organismal metabolic rate. Significant increases in energy use (inferred from blood flow) above that found in unloaded birds were found in 12 muscles in trunk loaded birds, with most of the increases restricted to stance-phase muscles, as predicted. Just three of these muscles, the femerotibialis, the iliotibialis lateralis pars postacetabularis and the fibularis longus accounted for 70% of the increased energy use. Noticeably absent from the group of muscles that increased energy use during trunk loading were several large biarticular muscles that have extensor actions at the hip or ankle, but flexor actions at the knee. We concluded that the low energetic cost of carrying trunk loads in guinea fowl may rely on the activation of a group of muscles that together provide support and propulsion across all the major joints, without producing opposing moments at other joints that could potentially waste energy. The specific leg muscles responsible for the increase in metabolism during trunk loading also suggest that the energy cost of producing mechanical work may be an important determinant of the cost of carrying extra mass on the trunk. During distal-limb loading, eleven leg muscles had significant increases in energy use, but unlike during trunk loading, both stance- and swing-phase muscles had large increases in energy use. This distribution of energy use between stance and swing agrees with the prediction that increased mechanical work determines the cost of limb loading, because a substantial fraction of the increased segmental work during distal-limb loading in guinea fowl has been found to occur during stance. PMID- 16709910 TI - Nutrient regulation in relation to diet breadth: a comparison of Heliothis sister species and a hybrid. AB - We examined the nutritional correlates of diet breadth in insect herbivores by comparing patterns of diet selection, nutrient balancing, post-ingestive utilization and development in two sister species of caterpillar and a hybrid between them. One species, Heliothis virescens (HV) has a broad host range, feeding on plants in at least 14 families. The other, Heliothis subflexa (HS), is a specialist on plants in the genus Physalis (Solanaceae). Experiments using synthetic foods showed that when the caterpillars were allowed to mix their diet, the generalist self-selected a higher-protein diet whereas the specialist ate almost equal amounts of protein and carbohydrate, which accords with differences between the two species in the nutrient content of their natural diets. When confined to nutritionally imbalanced diets, the generalist showed a propensity to over-eat high protein:carbohydrate (P:C) diets to a greater degree than did the specialist and maintained higher rates of development. The generalist did not, however, over-eat low P:C diets to the same degree as the specialist. The hybrid selected a diet composition that was indistinguishable to that of its generalist father (HV), while its response to imbalanced diets was closely similar to that of the specialist mother (HS). The generalist converted ingested nutrient to growth with lower efficiency than did the specialist and the hybrid. Our findings imply that different behavioural and physiological traits linked to nutrient regulation are under genetic control and are explicable in terms of the different life-histories, feeding ecologies and presumed levels of nutritional heterogeneity in the environments of the two insects. PMID- 16709911 TI - In vivo cranial suture function and suture morphology in the extant fish Polypterus: implications for inferring skull function in living and fossil fish. AB - This study describes the mechanical role that cranial sutures play in fish during feeding. The long-term goal of our work is to establish relationships between suture form and function, so that functional inferences can be made from suture morphology in fossil taxa. To this end, strain gauges were surgically implanted across selected sutures in the skull roof of four individuals of Polypterus endlicherii. After surgery, bone and suture strains during feeding were recorded along with high-speed video of the feeding events. Each trial was designated as a suction feeding or biting on prey trial, and neurocranial elevation, hyoid position and gape were quantified to aid in interpreting the strain data. The strains due to suction feeding are different from those observed during biting. Suction feeding results in a fairly stereotyped strain pattern, with the interfrontal and frontoparietal sutures experiencing tension, while the interparietal suture is compressed. Biting causes much more variable strain patterns. However, both suction and biting result in compression in the back of the skull, and tension between the frontals. Peak strains, and the time at which they occur in the feeding cycle, were compared between suction and biting. In general, peak suture strains are higher during suction than during biting, but not all of these differences are significant. Peak suture and bone strains occur at or near maximum gape during both suction and biting, suggesting that these strains are caused by muscle contraction involved in mouth opening and closing. Micro-computed tomography (microCT) scans of the experimental specimens indicate that the interfrontal and frontoparietal sutures, typically loaded in tension, are less interdigitated in cross section than the interparietal suture, which experiences compression. This is consistent with published correlations of suture form and function in mammals, where interdigitated sutures indicate compression and lack of interdigitation is associated with tension. PMID- 16709912 TI - Balancing the competing requirements of saltatorial and fossorial specialisation: burrowing costs in the spinifex hopping mouse, Notomys alexis. AB - Semi-fossorial animals (burrowing surface foragers) need to balance the competing morphological requirements of terrestrial and burrowing locomotion. These species rarely show the same degree of claw, forelimb and pectoral girdle structural development that fully fossorial forms (burrowing subterranean foragers) do, but nevertheless invest considerable amounts of energy in burrow systems. The compromise between terrestrial and burrowing locomotion was investigated by measuring net costs of burrowing and pedestrian transport in the spinifex hopping mouse, Notomys alexis, a species that forages in open areas in arid environments and is adapted for saltatorial locomotion. The net cost of transport by burrowing of hopping mice was found to be more expensive than for specialised fossorial species, and burrows were estimated to represent an energy investment equivalent to the terrestrial locomotion expected to be incurred in 17-100 days. A phylogenetically independent-contrasts approach revealed that morphological specialisation for burrowing was associated with low maximum running speeds in fossorial mammals and, for non-fossorial rodents and marsupials, maximum running speed was positively correlated with an index of habitat structure that ranged from arboreal to open desert. The high terrestrial speeds attainable by this semi fossorial species by saltatory locomotion apparently outweigh the energetic savings that would be associated with burrowing specialisation. PMID- 16709913 TI - Steroid-induced cardiac contractility requires exogenous glucose, glycolysis and the sarcoplasmic reticulum in rainbow trout. AB - Recent data from our laboratory suggest that sex steroids promote contractile function in cardiac muscle of rainbow trout (Oncorhynchus mykiss Walbaum), and there are sex differences in hormone signaling and cardiac function. The current study investigated whether steroid-induced inotropism in electrically paced (0.5 Hz, 14 degrees C) ventricle strips at 90% Lmax (1) has a metabolic requirement for exogenous glucose and (2) is associated with enhanced intracellular Ca2+ storage and release from the sarcoplasmic reticulum (SR). We also explored whether sex differences exist in extracellular Ca2+ (Ca2+o) or cardiac sensitivity to Ca2+o. In the absence or at low concentrations (1 or 2 mmol l-)) of exogenous glucose, resting tension and relaxation time were increased selectively in cardiac tissue from females. Increasing glucose promoted twitch force in a bell-shaped manner, with 5 mmol l-1 representing the optimal concentration for both sexes. The positive inotropic effects of physiological concentrations of testosterone (T) and 17beta-estradiol (E2) in male and female trout ventricle strips, respectively, developed slowly (10-45 min) and were not apparent in glucose-free medium, in medium containing iodoacetate (IAA), an inhibitor of glycolysis, or medium containing 5 mmol l-) lactate or pyruvate. Male ventricle strips had increased inotropic responses to glucose and T compared with female strips exposed to glucose and E2. Furthermore, sexually maturing males showed a greater inotropic response than immature males or females. Pretreatment with ryanodine (a specific blocker of SR Ca2+ release) also eliminated the inotropic effects of sex steroids and exogenous glucose and reduced the post-rest potentiation of contractile force (a marker of SR Ca2+ storage). By contrast, the inotropic effects of epinephrine (Epi) or elevated Ca2+o were faster (developing within 1-3 min) and were not diminished by the presence or absence of glucose or by pretreatment with IAA or ryanodine. Sex differences were also found in responsiveness to caffeine (males>females) and the relationship between Ca2+ concentration and force development above baseline. The Ca2+50 was lower in female cardiac tissue than males, suggesting greater Ca2+ sensitivity, and although plasma albumin was higher in females, total and ionized plasma Ca2+ did not differ between the sexes. For the first time, our study highlights the importance of extracellular glucose, glycolytic activity and SR Ca2+ storage and release for sex steroid-induced inotropism in the trout ventricle. Conversely, the inotropes Epi and elevated [Ca2+o] do not require the presence or metabolism of exogenous glucose or the SR for signaling their positive effects on contractility. These results also demonstrate novel sex related differences in cardiac reliance on exogenous glucose, Ca2+ sensitivity and SR function and thus should be considered in future studies. PMID- 16709914 TI - Hypoxia induces a complex response of globin expression in zebrafish (Danio rerio). AB - Unlike most mammals, many fish species live and survive in environments with low or changing levels of oxygen. Respiratory proteins like hemoglobin or myoglobin bind or store oxygen, thus enhancing its availability to the respiratory chain in the mitochondria. Here we investigate by means of quantitative real-time PCR the changes of hemoglobin, myoglobin, neuroglobin, cytoglobin and globin X mRNA in zebrafish (Danio rerio) exposed to mild (PO2=approximately 8.6 kPa) or severe (PO2=approximately 4.1 kPa) hypoxia. Neuroglobin and myoglobin protein levels were investigated by western blotting. Whereas mild hypoxia caused only minor changes of mRNA levels, strong hypoxia enhanced mRNA levels of the control genes (lactate dehydrogenase A and phosphoglycerate kinase 1). Surprisingly, levels of hemoglobin alpha and beta mRNA were significantly reduced under severe hypoxia. Myoglobin mRNA and protein in heart mildly increased, in line with its proposed oxygen supply function. Likewise, neuroglobin mRNA and protein significantly increased in brain (up to 5.7-fold at the protein level), but not in eye. This observation, firstly, suggests physiological differences of zebrafish eye and brain under hypoxia, and secondly, indicates an important role of neuroglobin in oxidative metabolism, probably oxygen supply within neurons. There was little change in the expression of the two cytoglobin genes. Globin X mRNA significantly decreased under hypoxia, pointing to a functional linkage to oxygen-dependent metabolism. PMID- 16709915 TI - Walking by Ixodes ricinus ticks: intrinsic and extrinsic factors determine the attraction of moisture or host odour. AB - Spontaneous walking by Ixodes ricinus nymphs in the horizontal plane was investigated within choice arenas to test whether these ticks move preferentially up a humidity gradient, and whether they are attracted towards host odour, specifically volatile substances from the back of a dog's ear. Responses by ticks with high and low fat contents and in different states of hydration were compared. Ticks with higher energy reserves were more likely to walk horizontally over short distances. Only if they were slightly dehydrated were they more likely to walk towards fully saturated air than drier air, and only if the atmosphere was sufficiently wet were they likely to walk towards odour secreted by host skin. It seems that, under certain circumstances, ticks of this ambushing species will move short distances towards host odour. PMID- 16709916 TI - The human gluteus maximus and its role in running. AB - The human gluteus maximus is a distinctive muscle in terms of size, anatomy and function compared to apes and other non-human primates. Here we employ electromyographic and kinematic analyses of human subjects to test the hypothesis that the human gluteus maximus plays a more important role in running than walking. The results indicate that the gluteus maximus is mostly quiescent with low levels of activity during level and uphill walking, but increases substantially in activity and alters its timing with respect to speed during running. The major functions of the gluteus maximus during running are to control flexion of the trunk on the stance-side and to decelerate the swing leg; contractions of the stance-side gluteus maximus may also help to control flexion of the hip and to extend the thigh. Evidence for when the gluteus maximus became enlarged in human evolution is equivocal, but the muscle's minimal functional role during walking supports the hypothesis that enlargement of the gluteus maximus was likely important in the evolution of hominid running capabilities. PMID- 16709917 TI - Involvement of ryanodine-operated channels in tert-butylhydroperoxide-evoked Ca2+ mobilisation in pancreatic acinar cells. AB - Reactive oxygen species and related oxidative damage have been implicated in the initiation of acute pancreatitis, a disease characterised in its earliest stages by disruption of intracellular Ca2+ homeostasis. The present study was carried out in order to establish the effect of the organic pro-oxidant, tert butylhydroperoxide (tBHP), on the mobilisation of intracellular Ca2+ stores in isolated rat pancreatic acinar cells and the mechanisms underlying this effect. Cytosolic free Ca2+ concentrations ([Ca2+]c) were monitored using a digital microspectrofluorimetric system in fura-2 loaded cells. In the presence of normal extracellular Ca2+ concentrations ([Ca2+]o), perfusion of pancreatic acinar cells with 1 mmol l-1 tBHP caused a slow sustained increase in [Ca2+]c. This increase was also observed in a nominally Ca2+-free medium, indicating a release of Ca2+ from intracellular stores. Pretreatment of cells with tBHP abolished the typical Ca2+ response of both the physiological agonist CCK-8 (1 nmol l-1) and thapsigargin (TPS, 1 micromol l-1), an inhibitor of the SERCA pump, in the absence of extracellular Ca2+. Similar results were observed with carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP, 0.5 micromol l-1), a mitochondrial uncoupler. In addition, depletion of either agonist-sensitive Ca2+ pools by CCK-8 or TPS or mitochondrial Ca2+ pools by FCCP were unable to prevent the tBHP-induced Ca2+ release. By contrast, simultaneous administration of TPS and FCCP clearly abolished the tBHP-induced Ca2+ release. These results show that tBHP releases Ca2+ from agonist-sensitive intracellular stores and from mitochondria. On the other hand, simultaneous application of FCCP and of 2 aminoethoxydiphenylborane (2-APB), a blocker of IP3-mediated Ca2+ release, was unable to suppress the increase in [Ca2+]c induced by tBHP, while the application of 50 micromol l-1 of ryanodine (which is able to block the ryanodine channels) inhibits tBHP-evoked Ca2+ mobilisation. These findings indicate that tBHP releases Ca2+ from non-mitochondrial Ca2+ pools through ryanodine channels. PMID- 16709918 TI - Evidence that blue petrel, Halobaena caerulea, fledglings can detect and orient to dimethyl sulfide. AB - Procellariiform seabirds (the petrels, albatrosses and shearwaters) are recognized for their acute sense of smell. These pelagic seabirds forage over thousands of miles of ocean to find patchily distributed prey resources. Over the past decade, much headway has been made in unravelling the variety of olfactory foraging strategies that Antarctic species employ, and it is becoming clearer that olfaction plays a key role in foraging, particularly for burrow nesting species. Now we are beginning to explore how these behaviours develop in chicks. Procellariiform chicks fledge and survive the open seas without aid or instruction from a parent, but how they are able to accomplish this task is unknown. Here we explore whether chicks leave the nest pre-tuned to olfactory cues necessary for foraging. In this study, we tested the hypothesis that blue petrel chicks (Halobaena caerulea) are able to detect and orient to a foraging cue (dimethyl sulphide, DMS) used by adults without ever having experienced this odour at sea. We first established that chicks could detect DMS at a biologically relevant concentration that they will later naturally encounter at sea (<10 pmol l-1). We then performed preference tests in a Y-maze on a group of birds 1-6 days before they fledged. Sixteen out of 20 fledglings preferred DMS (e.g. DMS+propylene glycol) to a ;control' odour (propylene glycol alone). Our results suggest that chicks can detect and may already recognize DMS as an orientation cue even before they leave the nest to forage for the first time. PMID- 16709919 TI - Hydrodynamic consequences of flexural stiffness and buoyancy for seaweeds: a study using physical models. AB - An upright posture in the water column may expose benthic marine organisms to faster flow higher in the water column than near the substratum, potentially increasing rates of mass exchange while also exposing the organisms to higher hydrodynamic forces. Benthic organisms maintain upright postures in the water column by one of two mechanisms, stiffness or buoyancy. Turbinaria ornata is a tropical macroalga that uses either buoyancy or flexural stiffness (EI), depending on its flow habitat. This study used physical models of T. ornata to compare the effect of different magnitudes of these two mechanisms on relative water velocity and hydrodynamic forces in both unidirectional and wavy flow. Models of the alga were constructed to span and exceed natural levels of T. ornata's EI and buoyancy. Models with high EI and high buoyancy maintained upright postures in both unidirectional flow and waves, and experienced higher forces than models with low EI and moderate or low buoyancy that deflected in the direction of water motion. In waves, buoyant models that were deflected by high velocity rebounded back into upright positions when the flow slowed. Non-buoyant, flexible models were also pushed over by flow but lacked the ability to rebound upright, which led to decreased force in unidirectional flow, but high force in waves. PMID- 16709920 TI - Time-domain signal divergence and discrimination without receptor modification in sympatric morphs of electric fishes. AB - Polymorphism in an animal communication channel provides a framework for studying proximate rules of signal design as well as ultimate mechanisms of signal diversification. Reproductively isolated mormyrid fishes from Gabon's Brienomyrus species flock emit distinctive electric organ discharges (EODs) thought to function in species and sex recognition. Species boundaries and EODs appear congruent in these fishes, with the notable exception of three morphs designated types I, II and III. Within the species flock, these morphs compose a monophyletic group that has recently been called the magnostipes complex. Co occurring morphs of this complex express distinctive EODs, yet they appear genetically indistinguishable at several nuclear loci. In this study, we investigated EOD discrimination by these morphs using both behavioral and physiological experiments. During the breeding season, wild-caught type I and type II males showed evidence that they can discriminate their own morph's EOD waveform from that of a sympatric and genetically distinct reference species. However, we found that type I and type II males exhibited an asymmetry in unconditioned responses to paired playback of EODs recorded from type I versus type II females. Males of the type II morph responded preferentially to EODs of type II females, whereas type I males did not appear to discriminate homotypic and heterotypic EODs in our experimental paradigm. Part of this behavioral asymmetry may have resulted from a previously undetected difference in adult size, which may have enhanced apparent discrimination by the smaller morph (type II) due to a relatively higher risk of injury from the larger morph (type I). Knollenorgan receptors, which mediate electrical communication in mormyrids, showed similar spectral tuning in type I and type II. These electroreceptors coded temporal features of any single magnostipes-complex EOD with similar patterns of time-locked spikes in both morphs. By contrast, Knollenorgans exhibited distinctive responses to different EOD waveforms. These results suggest that discrete EOD variation in this rapidly diversifying complex is functional in terms of morph-specific advertisement and recognition. Time-domain signal divergence has outpaced frequency-domain divergence between sympatric morphs, requiring little to no change in receptor response properties. We discuss our findings in light of a model for EOD time-coding by the Knollenorgan pathway, as well as evolutionary hypotheses concerning sympatric signal diversification in the magnostipes complex. PMID- 16709921 TI - Perturbation of leg protraction causes context-dependent modulation of inter-leg coordination, but not of avoidance reflexes. AB - All animals capable of legged locomotion execute fast, adaptive compensatory movements in response to perturbation of a step cycle. In terms of motor control, such adaptive behaviour typically involves changes in the kinematics of the perturbed limb as well as changes in coordination between legs. Moreover, the unpredictable variety of real life situations implies that compensatory responses should be sensitive to the behavioural context of the animal. We have investigated the extent to which the compensatory response of a walking stick insect (Carausius morosus) adapts in parallel to strong context-dependent adaptation of step kinematics and inter-leg coordination. The behavioural contexts we chose were straight walking and visually induced curve walking, for both of which the steady state limb kinematics and inter-leg coupling strengths were known. In case of curve walking, we further distinguished contexts according to whether the inner or the outer leg was perturbed. The three contexts differed strongly with respect to the set of joint actions before perturbation. Upon mechanical perturbation of front leg protraction, we studied context-dependent differences in a local avoidance reflex of the perturbed leg, as well as in coordination mechanisms that couple the step cycles of the perturbed leg to its unperturbed neighbours. In all three walking contexts, obstacle contact caused an avoidance movement of the front leg that deviated from the unperturbed swing trajectory. Swing duration was increased while step distance was decreased; however, both effects vanished in the subsequent unperturbed step. The prevailing immediate reaction of the three leg joints were retraction of the coxa (>76%), levation of the femur (>80%), and flexion of the tibia (>80%), regardless of the behavioural context and, therefore, joint action prior to perturbation. Moreover, activation of each one of these joint actions was shown to be independent of the other two. Thus, local avoidance reflexes are not modulated by the descending visual information that causes transition from straight to curve walking, but are composed of context-independent joint actions. Perturbation of the front leg also caused significant shifts of the touch-down position of the perturbed leg and of its unperturbed neighbours. If the inner front leg was perturbed, this shift could persist until the subsequent step. Perturbation affected both the spatial location and the timing of touch-down and lift-off transitions in unperturbed neighbouring legs. These effects on inter-leg coordination were context dependent. For example, time delay to lift-off of the contralateral neighbour was shortened in inner and straight walking legs, but not in outer legs. Finally, a targeting mechanism that determines foot placement in stick insects was shown to be affected by perturbation in a context-dependent manner. We conclude that the immediate compensatory response of the perturbed leg is not adapted to the behavioural context in spite of strongly differing step kinematics, whereas the compensatory effect on inter-limb coupling is context-dependent. PMID- 16709922 TI - The regenerative nidi of the locust midgut as a model to study epithelial cell differentiation from stem cells. AB - A better knowledge of the regulatory mechanisms involved in stem cell proliferation and/or differentiation could reveal new methods for the treatment of some diseases. Most previous studies in the field of stem cell biology have been carried out on cultured isolated cells. In the case of adult tissue stem cells, mesenchymal bone marrow derived cells have been most widely studied, while the undifferentiated stem cells present in the epithelial tissues are less known. In order to advance further our understanding of epithelial tissue stem cells, new in vivo models are required. The present study focuses on the dynamics of a new and simple model of intestinal epithelial regeneration found in the midgut of the migratory locust, Locusta migratoria (Linnaeus 1758). The locust midgut consists of three cell types: columnar cells, endocrine cells and undifferentiated regenerative clustered cells. The undifferentiated epithelial midgut cells give rise to two other cell types and are located in a nest of regenerative cells known as regenerative niche. We have performed single and continuous bromodeoxyuridine (BrdU) administration experiments to study regeneration niches and their cellular dynamics. Immunocytochemistry and immunofluorescence techniques were used to detect the incorporation of BrdU into regenerative niches and the presence of FMRFamide-like immunoreactivity, as a marker for endocrine cell differentiation. Some isolated label retaining cells (LRC) were observed at the niche base 10-15 days after the final BrdU administration. We propose that these cells are the stem cells of this epithelial tissue. We also calculated the length of the cell cycle phases for a subpopulation of transit amplifying cells within the regenerative niche: G1, 2.5+/-0.5 h; S, 5.5+/-0.5 h; G2, 0.75+/-0.25 h; M, 2.5+/-0.5 h. These amplifying cells will give rise to the columnar epithelial non-endocrine lineage. The differentiation of an endocrine cell from a niche stem cell occurs less frequently and thus leads to a lower proportion of endocrine cells as compared with epithelial columnar digestive cells (up to three endocrine cells per niche). Endocrine cell commitment seems to occur very early in the differentiation process within the niche, as double-labelled BrdU and FMRF endocrine cells have never been found. The only exception is the endocrine cells located in the ampullar region of the midgut, some of which show double immunostaining after long-term chronic BrdU injection. In summary, we have characterized a new and simple animal model of epithelial stem cell regeneration that may be useful for understanding the complex biological process that drives tissue renewal from undifferentiated and uncommitted progenitor cells. PMID- 16709923 TI - Locomotor function of the pectoral girdle 'muscular sling' in trotting dogs. AB - In therian mammals, gravitational and locomotor forces are transferred between the forelimb and trunk primarily, or entirely, through the muscles that connect the limb and trunk. Our understanding of this force transmission is based on analyses of shoulder anatomy and on a handful of descriptive electromyographic studies. To improve our understanding, we manipulated the locomotor forces of trotting dogs and monitored the resulting change in recruitment of five extrinsic muscles of the forelimb: m. serratus ventralis thoracis, m. serratus ventralis cervicis, m. pectoralis superficialis transversus, the anterior portion of the m. pectoralis profundus, and m. rhomboideus thoracis. Locomotor forces were modified as the dogs trotted at constant speed on a motorized treadmill by (1) adding mass to the trunk, (2) inclining the treadmill so that the dogs ran up and down hill, (3) adding mass to the wrists and (4) applying horizontally directed force to the trunk through a leash. These experiments indicate that the thoracic portion of the serratus ventralis muscle is the main antigravity muscle of the shoulder during trotting in dogs. Its activity increased when we added mass to the trunk and also when we ran the subjects downhill. In contrast, the cervical portion of the serratus ventralis did not show a consistent increase in activity in response to added mass. Instead, its activity increased when we ran the subjects up hill and added mass to their wrists, suggesting that it functions to stabilize the fulcrum of the forelimb in the cranial-caudal direction during active retraction of the forelimb. The thoracic portion of the rhomboideus muscle also appears to provide this cranial-caudal stabilization during active retraction of the forelimb. The force manipulations indicate that the transverse pectoralis muscle acts to both protract and retract the forelimb, depending on the position of the limb. In contrast, the anterior portion of the pectoralis profundus muscle acts as a retractor of the forelimb during the end of swing phase and the beginning of support phase. We found that adding mass to the trunk did not increase the activity of forelimb retractor muscles, suggesting that the ground reaction force vector passes through, or very near, the fulcrum of the shoulder during a trotting step. Whether or not the functions of these extrinsic appendicular muscles in dogs characterize therian mammals or represent specializations for high-speed, economical running remains to be determined. PMID- 16709924 TI - Suppression of T-cell functions by human granulocyte arginase. AB - Chronic inflammation is accompanied by impaired T-cell immunity. In the mouse, myeloid cell-associated arginase accounts for the suppression of immune reactivity in various models of tumor growth and chronic infections. Here we show that arginase I is liberated from human granulocytes, and very high activities accumulate extracellularly during purulent inflammatory reactions. Human granulocyte arginase induces a profound suppression of T-cell proliferation and cytokine synthesis. This T-cell phenotype is due to arginase-mediated depletion of arginine in the T-cell environment, which leads to CD3zeta chain down regulation but does not alter T-cell viability. Our study therefore demonstrates that human granulocytes possess a previously unanticipated immunosuppressive effector function. Human granulocyte arginase is a promising pharmacologic target to reverse unwanted immunosuppression. PMID- 16709925 TI - Induction of Fc gammaRIIA expression in myeloid PLB cells during differentiation depends on cytosolic phospholipase A2 activity and is regulated via activation of CREB by PGE2. AB - Fc gammaRIIA expressed on neutrophils and monocytes has a fundamental role in combating bacterial infections. In the present study, the requirement of cytosolic phospholipase A2 (cPLA2) for induction of Fc gammaRIIA expression was studied in a model of cPLA2-deficient PLB-985 cells (PLB-D cells). Fc gammaRIIA was acquired only during differentiation of PLB but not of PLB-D cells induced by either 1,25-dihydroxyvitamin D3, retinoic acid, or interferon gamma. Addition of prostaglandin E2 (PGE2) to PLB-D cells undergoing differentiation restored the expression of Fc gammaRIIA protein, whereas addition of indomethacin to PLB cells during differentiation inhibited both the production of PGE2 and the expression of Fc gammaRIIA. Inhibition of PKA during PLB differentiation prevented Fc gammaRIIA expression, whereas dibutyryl cAMP (dbcAMP) induced its expression in both PLB and PLB-D cells. CREB phosphorylation and CREB-CRE interaction were detected only in differentiated PLB cells and not PLB-D cells and were inhibited by indomethacin. A reporter gene containing a Fc gammaRIIA gene promoter fragment with the CRE element was sufficient for CREB activation. Our results are the first to show that CREB activation is involved in up-regulation of Fc gammaRIIA expression in myeloid lineages. PGE2 formed via cPLA2 activates CREB through PKA and this process is dependent on development of PGE2 receptor 4. PMID- 16709926 TI - Aire-deficient mice develop hematopoetic irregularities and marginal zone B-cell lymphoma. AB - Autoimmune polyendocrine syndrome type I (APS I) is an inherited recessive disorder with a progressive immunological destruction of many tissues including the adrenal cortex, the parathyroid glands, and the gonads. APS I is caused by mutations in the AIRE gene (autoimmune regulator), expressed in cells of the thymus and spleen, suggesting a role in central and peripheral tolerance. Aire(-/ ) mice replicate the autoimmune features of APS I patients with the presence of multiple autoantibodies and lymphocytic infiltrates in various tissues, but young mice appear clinically healthy. We here report the investigation of 15- to 24 month-old Aire(-/-) mice. We did not observe any endocrinological abnormalities, nor did sera from these mice recognize known APS I autoantigens. Interestingly, however, there was a high frequency of marginal zone B-cell lymphoma in Aire(-/-) mice and liver infiltrates of B cells, suggesting chronic antigen exposure and exaggerated activation. Furthermore, increased numbers of monocytes in blood were identified as well as augmented numbers of metallophilic macrophages in the spleen. We propose that Aire, in addition to its function in the thymus, also has a peripheral regulatory role by controlling the development of antigen-presenting cells (APCs) and marginal zone B-cell activation. PMID- 16709927 TI - Signaling of vascular endothelial growth factor receptor-1 tyrosine kinase promotes rheumatoid arthritis through activation of monocytes/macrophages. AB - Vascular endothelial growth factor (VEGF) and VEGF receptor-1 (VEGFR-1/Flt-1) were shown to be involved in pathological angiogenesis, particularly rheumatoid arthritis (RA). However, the molecular basis of their actions is not fully understood. Here we report that in a murine model of RA, deletion of the tyrosine kinase (TK) domain of VEGFR-1 decreased the incidence and clinical symptoms of RA. Pathological symptoms, such as synovial hyperplasia, inflammatory infiltrates, pannus formation, and cartilage/bone destruction, became milder in Vegfr-1 tk(-/-) mice compared with wild-type (Wt) mice in the human T-cell leukemia virus-1 (HTLV-1) pX-induced chronic models. VEGFR-1 TK-deficient bone marrow cells showed a suppression of multilineage colony formation. Furthermore, macrophages induced to differentiate in vitro showed a decrease in immunologic reactions such as phagocytosis and the secretion of interleukin-6 (IL-6) and VEGF A. Treatment of this RA model with a small molecule inhibitor for VEGFR TK, KRN951, also attenuated the arthritis. These results indicate that the VEGFR-1 TK signaling modulates the proliferation of bone marrow hematopoietic cells and immunity of monocytes/macrophages and promotes chronic inflammation, which may be a new target in the treatment of RA. PMID- 16709928 TI - Expression of the outcome predictor in acute leukemia 1 (OPAL1) gene is not an independent prognostic factor in patients treated according to COALL or St Jude protocols. AB - New prognostic factors may result in better risk classification and improved treatment of children with acute lymphoblastic leukemia (ALL). Recently, high expression of a gene named OPAL1 (outcome predictor in acute leukemia) was reported to be associated with favorable prognosis in ALL. Therefore, we investigated whether OPAL1 expression was of prognostic importance in 2 independent cohorts of children with ALL treated on Cooperative Study Group for Childhood Acute Lymphoblastic Leukemia (COALL)-92/97 (n = 180) and St Jude Total 13 protocols (n = 257). We observed a consistently higher (2.8-fold) expression of OPAL1 in TEL-AML1-positive ALL compared with TEL-AML1-negative ALL in both cohorts, but higher OPAL1 expression was not consistently associated with other favorable prognostic indicators such as age and white blood cell count, or ALL genetic subtype. Lower OPAL1 expression was also not associated with increased in vitro drug resistance. Multivariate analyses including known risk factors showed that OPAL1 expression was not independently related to prognosis in either the COALL or St Jude cohorts. In conclusion, OPAL1 expression may not be an independent prognostic feature in childhood ALL, and its previously reported prognostic impact appears to be treatment dependent. PMID- 16709929 TI - High molecular response rate of polycythemia vera patients treated with pegylated interferon alpha-2a. AB - V617F JAK2 mutation is a reliable molecular marker of polycythemia vera (PV), potentially useful to monitor the effect of treatments in this disease. In a phase 2 study of pegylated (peg) IFN-alpha-2a in PV, we performed prospective sequential quantitative evaluation of the percentage of mutated JAK2 allele (%V617F) by real-time polymerase chain reaction (PCR). The %V617F decreased in 24 (89%) of 27 treated patients, from a mean of 49% to a mean of 27% (mean decrease of 44%; P < .001), and no evidence for a plateau was observed. In one patient, mutant JAK2 was no longer detectable after 12 months. In 3 patients homozygous for the mutation, reappearance of 50% of wild-type allele was observed during treatment. The results seem to confirm the hypothesis that IFN-alpha preferentially targets the malignant clone in PV and show that %V617F assessment using a quantitative method may provide the first tool to monitor minimal residual disease in PV. This trial was registered at www.clinicaltrials.gov as #NCT00241241. PMID- 16709930 TI - Evolving concepts in the management of chronic myeloid leukemia: recommendations from an expert panel on behalf of the European LeukemiaNet. AB - The introduction of imatinib mesylate (IM) has revolutionized the treatment of chronic myeloid leukemia (CML). Although experience is too limited to permit evidence-based evaluation of survival, the available data fully justify critical reassessment of CML management. The panel therefore reviewed treatment of CML since 1998. It confirmed the value of IM (400 mg/day) and of conventional allogeneic hematopoietic stem cell transplantation (alloHSCT). It recommended that the preferred initial treatment for most patients newly diagnosed in chronic phase should now be 400 mg IM daily. A dose increase of IM, alloHSCT, or investigational treatments were recommended in case of failure, and could be considered in case of suboptimal response. Failure was defined at 3 months (no hematologic response [HR]), 6 months (incomplete HR or no cytogenetic response [CgR]), 12 months (less than partial CgR [Philadelphia chromosome-positive (Ph(+)) > 35%]), 18 months (less than complete CgR), and in case of HR or CgR loss, or appearance of highly IM-resistant BCR-ABL mutations. Suboptimal response was defined at 3 months (incomplete HR), 6 months (less than partial CgR), 12 months (less than complete CgR), 18 months (less than major molecular response [MMolR]), and, in case of MMolR loss, other mutations or other chromosomal abnormalities. The importance of regular monitoring at experienced centers was highlighted. PMID- 16709932 TI - Inhibition of RhoA GTPase activity enhances hematopoietic stem and progenitor cell proliferation and engraftment. AB - Ras-related Rho GTPases regulate actin cytoskeletal organization, adhesion, gene transcription, and cell-cycle progression. The Rac subfamily of Rho GTPases and Cdc42 has been shown to play essential roles in hematopoietic stem cell (HSC) engraftment and mobilization. Here, we study the role of RhoA, a related Rho GTPase, in HSC functions. Using retrovirus-mediated gene transfer of a dominant negative (DN) mutant of RhoA (RhoAN19), we demonstrate that down-regulation of RhoA activity resulted in increased HSC engraftment and self-renewal as measured by competitive repopulation and serial transplantation assays. However, overexpression of RhoAN19 resulted in decreased migration toward SDF-1alpha and alpha(4)beta(1)- and alpha(5)beta(2)-integrin-mediated adhesion of hematopoietic progenitor cells in vitro. Low RhoA activity was associated with higher proliferation rate of hematopoietic progenitor cells and increased cells in active phases of cell cycle, most likely via decreasing p21Cip/Waf expression and increasing cyclin D1 levels. Thus, reducing RhoA activity by optimizing the balance between adhesion/migration and proliferation/self-renewal results in a net increase in HSC engraftment. This mechanism could provide a novel therapeutic target to enhance HSC therapies. PMID- 16709931 TI - Survival benefit with imatinib mesylate versus interferon-alpha-based regimens in newly diagnosed chronic-phase chronic myelogenous leukemia. AB - A survival benefit for imatinib mesylate versus interferon-alpha therapy could not be demonstrated in the randomized study in newly diagnosed Philadelphia chromosome (Ph)-positive chronic-phase chronic myelogenous leukemia (CML) due to the high rate of crossover (90%) from interferon-alpha to imatinib mesylate within a year of study entry. We compared survival in 279 patients with newly diagnosed CML treated with imatinib mesylate at our institution (2000-2004) to 650 patients treated with interferon-alpha (1982-1997). The complete cytogenetic response rates were 87% with imatinib mesylate and 28% with interferon-alpha (P < .001). The estimated 3-year survival rates were 96% with imatinib mesylate and 81% with interferon-alpha (P < .01). Survival rates with imatinib mesylate were significantly better than with interferon-alpha within each of the CML prognostic risks groups. By multivariate analysis, imatinib mesylate therapy was identified as an independent favorable prognostic factor, after accounting for the impact of pretreatment factors (hazard ratio, 0.44; P < .01). By landmark analysis at 12 months, survival within each cytogenetic response category was similar with imatinib mesylate or interferon-alpha, suggesting that the survival benefit of imatinib mesylate (versus interferon-alpha in newly diagnosed CML) is through improving cytogenetic response. PMID- 16709933 TI - NPM-ALK-dependent expression of the transcription factor CCAAT/enhancer binding protein beta in ALK-positive anaplastic large cell lymphoma. AB - CCAAT/enhancer binding protein beta (C/EBPbeta) is one of a 6-member family of C/EBPs. These transcription factors are involved in the regulation of various aspects of cellular growth and differentiation. Although C/EBPbeta has important functions in B- and T-cell differentiation, its expression has not been well studied in lymphoid tissues. We, therefore, analyzed its expression by immunohistochemistry and Western blot in normal lymphoid tissues and in 248 well characterized lymphomas and lymphoma cell lines. Nonneoplastic lymphoid tissues and most B-cell, T-cell, and Hodgkin lymphomas lacked detectable levels of C/EBPbeta. In contrast, most (40 of 45; 88%) cases of ALK-positive anaplastic large cell lymphoma (ALCL) strongly expressed C/EBPbeta. Western blot analysis confirmed C/EBPbeta expression in the ALK-positive ALCLs and demonstrated elevated levels of the LIP isoform, which has been associated with increased proliferation and aggressiveness in carcinomas. Transfection of Ba/F3 and 32D cells with NPM-ALK and a kinase-inhibitable modified NPM-ALK resulted in the induction of C/EBPbeta and demonstrated dependence on NPM-ALK kinase activity. In conclusion, we report the constitutive expression of C/EBPbeta in ALK-positive ALCL and show its relationship to NPM-ALK. We suggest that C/EBPbeta is likely to play an important role in the pathogenesis and unique phenotype of this lymphoma. PMID- 16709934 TI - Clusterin decreases oxidative stress in lung fibroblasts exposed to cigarette smoke. AB - RATIONALE: Cigarette smoke causes injury to lung fibroblasts, partly by means of oxidative stress, and oxidative stress can lead to various lung diseases, such as chronic obstructive pulmonary disease. Clusterin is a widely distributed protein with many functions, including cellular protection in response to oxidative stress. OBJECTIVES: To determine whether clusterin is involved in the defense of the lung against cigarette smoke, we investigated the effects of cigarette smoke extract on clusterin expression and its protective effect, if any, against oxidative stress. METHODS: Fibroblasts were coincubated with conditioned medium and cigarette smoke extract, and bronchial biopsy specimens obtained from nonsmokers, smokers, and ex-smokers were analyzed by immunohistochemistry. MEASUREMENTS AND MAIN RESULTS: At concentrations of 2.5 and 5.0%, cigarette smoke extract induced oxidative stress. It also markedly increased the expression of two clusterin isoforms (60 and 76-80 kD) and the 76-80-kD isoform was secreted in the incubation medium. Coincubation of fibroblasts with conditioned medium significantly decreased the cellular oxidation caused by the cigarette smoke extract. Immunohistochemical analysis of clusterin on bronchial biopsy specimens obtained from smokers and ex-smokers showed localization of clusterin mainly in the submucosa. CONCLUSIONS: We conclude that clusterin may have a protective effect against cigarette smoke-induced oxidative stress in lung fibroblasts. PMID- 16709935 TI - Lack of weight gain and relapse risk in a large tuberculosis treatment trial. AB - BACKGROUND: Readily identified markers of tuberculosis relapse risk are needed, particularly in resource-limited settings. The association between weight gain or loss during antituberculosis therapy and relapse has not been well studied. METHODS: Subjects in the Tuberculosis Trials Consortium Study 22 were studied. Underweight was defined as 10% or more below ideal body weight at diagnosis. Weight change was assessed between (1) diagnosis and completion of induction phase therapy, (2) diagnosis and end of continuation phase therapy, and (3) completion of induction to completion of continuation phase therapy. RESULTS: A total of 857 subjects were monitored for 2 yr, and 61 of 857 (7.1%) relapsed. Relapse risk was high among persons who were underweight at diagnosis (19.1 vs. 4.8%; p < 0.001) or who had a body mass index of less than 18.5 kg/m(2) (19.5 vs. 5.8%; p < 0.001). Among persons who were underweight at diagnosis, weight gain of 5% or less between diagnosis and completion of 2-mo intensive phase therapy was moderately associated with an increased relapse risk (18.4 vs. 10.3%; relative risk, 1.79, 95% confidence interval, 0.96-3.32; p = 0.06). In a multivariate logistic regression model that was adjusted for other risk factors, a weight gain of 5% or less between diagnosis and completion of 2-mo intensive phase therapy among persons underweight at diagnosis was significantly associated with relapse risk (odds ratio, 2.4; p = 0.03). CONCLUSIONS: Among persons underweight at diagnosis, weight gain of 5% or less during the first 2 mo of treatment is associated with an increased relapse risk. Such high-risk patients can be easily identified, even in resource-poor settings. Additional studies are warranted to identify interventions to decrease risk of relapse in such patients. PMID- 16709936 TI - Extracellular matrix regulates enhanced eotaxin expression in asthmatic airway smooth muscle cells. AB - RATIONALE: Altered airway smooth muscle (ASM) function and enrichment of the extracellular matrix (ECM) with fibronectin and collagen are key features of asthma. Previously, we have reported these ECM proteins enhance ASM synthetic function. OBJECTIVE: We compared ASM cultured from endobronchial biopsies from subjects with and without asthma to assess if asthmatic cells were hypersecretory and determined whether the underlying mechanism involved autocrine ECM production. METHODS AND MEASUREMENTS: Cells from subjects with and without asthma were cultured on plastic or in plates precoated with ECM proteins. Cytokine production was evaluated by enzyme-linked immunosorbent assay and by reverse transcriptase-polymerase chain reaction. Function-blocking integrin antibodies were used to identify integrin involvement. RESULTS: Baseline eotaxin and its production after stimulation with interleukin (IL)-13, IL-1beta, or tumor necrosis factor-alpha was increased (2.5- to 6.0-fold) in ASM cells cultured from subjects with asthma compared with healthy subjects. When seeded on ECM from asthmatic ASM, IL-13-dependent eotaxin release from healthy or asthmatic ASM was enhanced compared with culture on healthy ECM. The ECM substrates fibronectin and type I collagen each enhanced IL-13-dependent eotaxin release, and Western immunoblot indicated that fibronectin expression was higher in asthmatic ASM cells. Integrin-blocking antibodies revealed that alpha5beta1 was required for more than 50% of the enhanced IL-13-dependent eotaxin release by ASM cells from subjects with asthma, whereas alpha2beta1 or alphavbeta3 neutralization lacked effect. CONCLUSION: The data indicate that ASM cells cultured from subjects with asthma are hypersecretory compared with cells from healthy donors and that autocrine fibronectin secretion acting via alpha5beta1 in part underlies this effect. PMID- 16709937 TI - Environmental tobacco smoke suppresses nuclear factor-kappaB signaling to increase apoptosis in infant monkey lungs. AB - RATIONALE: Exposure to environmental tobacco smoke in early life has adverse effects on lung development. Apoptosis plays an essential role in development; however, the molecular mechanisms of pulmonary apoptosis induced by environmental tobacco smoke is unknown. OBJECTIVES: To investigate the mechanistic role of nuclear factor (NF)-kappaB, a critical cell survival pathway, in the developing lungs exposed to environmental tobacco smoke. METHODS: Timed-pregnant rhesus monkeys and their offspring were exposed to filtered air or to aged and diluted sidestream cigarette smoke as a surrogate to environmental tobacco smoke (a total suspended particulate concentration of 0.99 mg/m(3) for 6 h/d, 5 d/wk) from 45-50 d gestational age to 72-77 d postnatal age (n = 4/group). MEASUREMENTS AND MAIN RESULTS: NF-kappaB-DNA binding activity, regulated anti-apoptotic genes, and apoptosis were measured in lung tissues. Exposure to environmental tobacco smoke significantly suppressed NF-kappaB activation pathway and activity. Environmental tobacco smoke further down-regulated NF-kappaB-dependent anti-apoptotic genes and induced activation of caspases, cleavage of cellular death substrates (poly(ADP) ribose polymerase and caspase-activated DNase) and an increase in the rate of apoptosis in the lung parenchyma. No significant alterations were observed for activator protein 1, p53 or Akt activity. CONCLUSIONS: Our results indicate that exposure to low levels of environmental tobacco smoke during a critical window of maturation in the neonatal nonhuman primate may compromise lung development with potential implications for future lung growth and function. These findings support our hypothesis that NF-kappaB plays a key role in the regulation of the apoptotic process. PMID- 16709939 TI - Variation in the human matrix metalloproteinase-9 gene is associated with arterial stiffness in healthy individuals. AB - BACKGROUND: Arterial stiffness is an important determinant of cardiovascular risk. Elastin is the main elastic component of the arterial wall and can be degraded by a number of enzymes including serine proteases and matrix metalloproteinases (MMPs). Serum MMP-9 levels correlate with arterial stiffness and predict cardiovascular risk. Polymorphisms in the MMP-9 gene are also associated with large artery function in subjects with coronary artery disease. Therefore, we investigated the influence of known MMP-9 (-1562C>T, R279Q) polymorphisms on arterial stiffness in a large cohort of healthy individuals (n=865). METHODS AND RESULTS: Aortic pulse wave velocity (PWV) and augmentation index were assessed. Supine blood pressure, biochemical markers, MMP-9 levels, and serum elastase activity (SEA) were also determined. Genomic DNA was extracted and genotyping performed. Aortic PWV, serum MMP-9, and SEA were higher in carriers of the rare alleles for the -1562C>T and R279Q polymorphisms. These polymorphisms were also associated with aortic PWV after correction for other confounding factors. Stepwise regression models with known or likely determinants of arterial stiffness revealed that approximately 60% of the variability in aortic PWV was attributable to age, mean arterial pressure, and genetic variants (P<0.001). CONCLUSIONS: We have demonstrated for the first time that aortic stiffness and elastase activity are influenced by MMP-9 gene polymorphisms. This suggests that the genetic variation in this protein may be involved in the process of large artery stiffening. PMID- 16709938 TI - Follow-up on metabolic markers in children treated for obstructive sleep apnea. AB - RATIONALE: In adults, obstructive sleep apnea (OSA) is associated with metabolic dysfunction that improves with treatment of OSA. No equivalent studies exist in children. OBJECTIVE: To examine the relationship between metabolic markers and OSA with time and treatment in children. METHODS: Metabolic markers measured on a fasting morning blood sample at diagnostic polysomnography and follow-up 1.3 +/- 0.6 yr later. MEASUREMENTS AND MAIN RESULTS: Forty-five children (34 males), aged 6.9 +/- 3.5 yr, and including 12 obese subjects, were in the final analysis. There were no differences in metabolic markers between children with and without OSA at initial study; however, obese children had significantly higher insulin (106.1 +/- 72.1 vs. 66.7 +/- 37.6 pmol/L; p = 0.028), insulin/glucose ratio (23.7 +/- 14.3 vs. 14.7 +/- 8.0; p = 0.02), and significantly lower high-density lipoprotein cholesterol (1.3 +/- 0.2 vs. 1.6 +/- 0.4 nmol/L; p = 0.005) than nonobese children. Twenty children underwent surgical removal of adenotonsillar tissue, whereas 12 children with OSA elected not to have treatment. OSA persisted after treatment in five children, and resolved in 27. Thirteen children did not have OSA on initial or follow-up studies. At follow-up, there was a small but significant improvement in total cholesterol in those children whose OSA was resolved (4.8 +/- 0.8 to 4.7 +/- 0.6 nmol/L; p = 0.005) and a trend for obese children with persisting OSA to have elevated insulin levels compared with obese children without OSA (p = 0.07). CONCLUSION: Obesity appears to be the major influence on metabolic dysfunction in children with OSA, but these preliminary data also suggest that resolution or persistence of OSA may affect changes in metabolic function over time. PMID- 16709941 TI - Augmentation of proliferation of vascular smooth muscle cells by plasminogen activator inhibitor type 1. AB - OBJECTIVE: Proliferation of vascular smooth muscle cells (VSMCs) contributes to restenosis after coronary intervention. We have shown previously that increased expression of plasminogen activator inhibitor type 1 (PAI-1) limits VSMC apoptosis. Because apoptosis and proliferation appear to be linked, we sought to determine whether increased PAI-1 would affect VSMC proliferation. METHODS AND RESULTS: VSMCs were explanted from control and transgenic mice (SM22-PAI+) in which VSMC expression of PAI-1 was increased. Increased growth of SM22-PAI+-VSMCs (2.3+/-0.4-fold) reflected, at least partially, increased proliferation. Greater expression of FLICE-like inhibitory protein (FLIP; 2.7-fold) and its cleaved active form were seen in SM22-PAI+-VSMCs. The balance between caspase-8 and FLIP favored proliferation in SM22-PAI+-VSMCs. Increased expression of NF-kappaB and activation of extracellular signal-regulated kinase (ERK) were demonstrated in SM22-PAI+-VSMCs (fold=NF-kappaB=2.2+/-0.1, fold=phosphorylated-ERK=1.6+/-0.1). Results were confirmed when expression of PAI-1 was increased by transfection. Inhibition of NF-kappaB and ERK attenuated proliferation in SM22-PAI+-VSMCs. Increased expression of PAI-1 promoted proliferation when VSMCs were exposed to tumor necrosis factor (TNF). CONCLUSIONS: Increased expression of PAI-1 is associated with greater activity of FLIP that promotes VSMC proliferation through NF-kappaB and ERK. Thus, when vascular wall expression of PAI-1 is increased, restenosis after coronary intervention is likely to be potentiated by greater proliferation of VSMC and resistance to apoptosis. PMID- 16709940 TI - Inhibition of fibroblast growth factor receptor signaling attenuates atherosclerosis in apolipoprotein E-deficient mice. AB - OBJECTIVE: To determine the significance of fibroblast growth factor receptor (FGFR) expression for the development of atherosclerotic lesions in apoE deficient (apoE-/-) mice. METHODS AND RESULTS: ApoE-/- mice fed a high-fat diet were administered the FGFR tyrosine kinase inhibitor SU5402 (25 mg/kg/d sc), which inhibited neointima growth by 85%. We measured its effects on lesion size at the aortic sinus, macrophage and smooth muscle cell (SMC) accumulation, the expression of monocyte chemotactic and retention factors, as well as its effects on FGFR expression/phosphorylation. FGFR tyrosine kinase inhibition reduced phosphorylated FGFRs in lesions by 90%, associated with a 65% reduction in lesion size measured using Oil Red O. Macrophages and SMCs within lesions were reduced by 58% and 78%, respectively. Monocyte chemotactic protein-1 (MCP-1) expression was also reduced, as was the expression of hyaluronan synthase, cyclooxygenase-2, CD36, and endothelial monocyte-activating polypeptide-II. Although 3 FGFR types were expressed in lesions, the effects of SU5402 could be attributed largely to inhibition of FGFR-1 phosphorylation. CONCLUSIONS: Atherosclerotic lesions in apoE-/- mice express multiple FGFRs and an active FGF:FGFR-1 signaling system that promotes atherosclerosis development via increased SMC proliferation, and by augmenting macrophage accumulation via increased expression of MCP-1 and factors promoting macrophage retention in lesions. PMID- 16709942 TI - Pathogenesis of calcific aortic valve disease: a disease process comes of age (and a good deal more). AB - BACKGROUND: Over the past 10 to 15 years, calcific aortic valve disease, which includes aortic sclerosis and aortic stenosis, has come to be recognized as an active process, based on: (1) epidemiologic studies demonstrating associations of specific risk factors with increased prevalence or rate of progression of aortic valve disease; (2) identification, in valve lesions, of histopathologic features of chronic inflammation, lipoprotein deposition, renin-angiotensin system components, and molecular mediators of calcification; and (3) identification of cell-signaling pathways and genetic factors that may participate in valve disease pathogenesis. These studies will be reviewed and organized into a proposed global hypothesis for the pathogenesis of calcific aortic valve disease. PMID- 16709943 TI - Familial elevated factor VIII in children with symptomatic venous thrombosis and post-thrombotic syndrome: results of a multicenter study. AB - OBJECTIVE: To evaluate the role of factor (F) VIII in children with non-cancer related venous thrombosis (DVT), post-thrombotic syndrome (PTS) or recurrent DVT. METHODS AND RESULTS: FVIII levels were measured in White patients and age- and gender-matched healthy controls. Heritability of factor VIII was estimated in 99 pedigrees by the variance component method implemented in SOLAR. The group of 103 patients showed higher median values of FVIII than 206 controls [FVIII:Ag, 115 versus 96 IU/dL, P<0.0001; FVIII:C, 119 versus 106 IU/dL, P=0.0009], and had a significantly increased odds ratio (OR) for fibrinogen-adjusted elevated FVIII levels [FVIII >90th percentile versus values below the cut-off: FVIII:Ag, OR 4.3, 95% confidence interval (CI) 1.5 to 12.1; FVIII:C, OR 5.5, CI 2.03 to 15.06]. PTS occurred in 19 of 59 children and persisted in 5 individuals. Recurrent DVT was seen in 8 patients. The heritable(h2)/household(c2) components were calculated for FVIII:Ag levels (h2, 0.48+/-0.15, P=0.0008; c2, 0.21), and FVIII:C (h2, 0.61+/-0.15, P<0.0001; c2, 0.41). When incorporating h2 and c2 in the estimate, the phenotypic variance for FVIII:Ag levels is predominantly explained by h2, whereas c2 stayed significant in the model for FVIII:C (P=0.00002). CONCLUSIONS: Elevated FVIII levels increase the DVT-risk in children. PMID- 16709944 TI - Reciprocal and coordinate regulation of serum amyloid A versus apolipoprotein A-I and paraoxonase-1 by inflammation in murine hepatocytes. AB - OBJECTIVE: During inflammation, the serum amyloid A (SAA) content of HDL increases, whereas apolipoprotein A-I (apoA-I) and paraoxonase-1 (PON-1) decrease. It remains unclear whether SAA physically displaces apoA-I or if these changes derive from coordinated but inverse transcriptional regulation of the HDL apolipoprotein genes. Because cytokines stimulate the hepatic expression of inflammatory markers, we investigated their role in regulating SAA, apoA-I, and PON-1 expression. METHODS AND RESULTS: A cytokine mixture (tumor necrosis factor [TNF]-alpha, interleukin [IL]-1beta, and IL-6) simultaneously induced SAA and repressed apoA-I and PON-1 expression levels. These effects were partially inhibited in cells pretreated with either nuclear factor kappaB (NF-kappaB) inhibitors (pyrrolidine dithiocarbamate, SN50, and overexpression of super repressor inhibitor kappaB) or after exposure to the peroxisome proliferator activated receptor-alpha (PPARalpha) ligands (WY-14643 and fenofibrate). Consistent with these findings, the basal level of SAA was increased, whereas apoA-I and PON-1 decreased in primary hepatocytes from PPARalpha-deficient mice as compared with wild-type mice. Moreover, neither WY-14643 nor fenofibrate had any effect on SAA, apoA-I, or PON-1 expression in the absence of PPARalpha. CONCLUSIONS: These results suggest that cytokines increase the expression of SAA through NF-kappaB transactivation, while simultaneously decreasing the expression of apoA-I and PON-1 by inhibiting PPARalpha activation. Inflammation may convert HDL de novo into a more proatherogenic form by coordinate but inverse transcriptional regulation in the liver, rather than by physical displacement of apoA-I by SAA. PMID- 16709945 TI - Laminar shear stress inhibits cathepsin L activity in endothelial cells. AB - OBJECTIVE: The cysteine proteases, cathepsins, have been implicated in vascular remodeling and atherosclerosis, processes known to be regulated by shear stress. It is not known, however, whether shear regulates cathepsins. We examined the hypothesis that shear stress regulates cathepsin activity in endothelial cells. METHODS AND RESULTS: Mouse aortic endothelial cells (MAECs) exposed to atheroprotective, unidirectional laminar shear (LS) degraded significantly less BODIPY-labeled elastin and gelatin in comparison to static and proatherogenic oscillatory shear (OS). The cathepsin inhibitor E64 also reduced this activity. Gelatin zymography showed that cathepsin activity of MAECs was blunted by LS exposure and by a cathepsin L inhibitor but not by cathepsin B and S inhibitors, whereas a cathepsin K inhibitor had a minor effect. Cathepsin L siRNA knocked down cathepsin L expression, gelatinase, and elastase activity in OS and static MAECs. A partial reduction of cathepsin B protein raised the possibility that the siRNA effect on the matrix protease activity could have been attributable to cathepsin L or B. Cathepsin B activity study using the synthetic peptide showed it was not regulated by shear. CONCLUSIONS: These results suggest that cathepsin L is a shear-sensitive matrix protease and that it may play an important role in flow-mediated vascular remodeling and atherogenic responses. PMID- 16709946 TI - Role of oxidative stress in remodeling of the myocardial microcirculation in hypertension. AB - OBJECTIVE: We tested the hypothesis that in early hypertension (HT), increased oxidative stress leads to myocardial microvascular remodeling. METHODS AND RESULTS: Pigs were studied after a 12-week observation: normal (n=8), untreated renovascular HT (n=8), or HT+chronic antioxidant supplementation (HT+A, n=6). Left ventricular muscle mass (LVMM) and myocardial blood flow (MBF) reserve were determined using electron beam computer tomography (CT), and the spatial density and tortuousity of myocardial microvessels (<500 microm) was then measured in myocardial samples with micro-CT. Myocardial microvascular morphology, oxidative stress, inflammation, and growth factor expression were determined in vitro. HT and HT+A had similarly increased arterial pressure and LVMM, but only HT showed impaired MBF response to adenosine. Compared with normal, HT had increased spatial density of myocardial microvessels, which was preserved in HT+A (111.8+/ 7.8, 166.3+/-15.7, and 106.4+/-6.1 vessels per cm2, respectively). HT also showed microvascular wall thickening, increased systemic and tissue oxidative stress, inflammation, and expression of vascular endothelial growth factor and its receptor Flk-1, most of which were attenuated by antioxidants. CONCLUSIONS: Myocardial microvascular remodeling in early HT is accompanied by tissue oxidative stress, inflammation, and altered growth factor expression, and attenuated by antioxidant intervention. This study underscores a role of increased oxidative stress in modulating myocardial microvascular architecture in early HT. PMID- 16709947 TI - Longitudinal differences in familial combined hyperlipidemia quantitative trait loci. PMID- 16709948 TI - Longitudinal differences in familial combined hyperlipidemia quantitative trait loci. PMID- 16709949 TI - Ruptures of delight? A new mouse model of plaque rupture. PMID- 16709950 TI - Plasma fibronectin concentration: a risk factor for arterial thrombosis? PMID- 16709951 TI - Maintenance of vascular homeostasis by bone marrow-derived cells. PMID- 16709952 TI - Insight into ABCG1-mediated cholesterol efflux. PMID- 16709953 TI - Lipoprotein metabolism: a well-tried tool to characterize dyslipidemic mechanisms. PMID- 16709954 TI - Lipoxygenase pathways as mediators of early inflammatory events in atherosclerosis. PMID- 16709957 TI - Rosiglitazone cools down inflammation in the metabolic syndrome. PMID- 16709958 TI - Activation of alveolar macrophages via the alternative pathway in herpesvirus induced lung fibrosis. AB - The etiology of idiopathic pulmonary fibrosis (IPF) is unknown. Because viral pathogenesis of IPF has been suggested, we have established a murine model of progressive pulmonary fibrosis by infecting IFN-gammaR-deficient mice (IFN gammaR(-/-)) with the murine gamma-herpesvirus 68. Because alveolar macrophages in humans with IPF have been implicated in driving the profibrotic response, we studied their role in our model. Chronic herpesvirus infection of the lung was associated with recruitment of alveolar macrophages to areas with epithelial hyperplasia and fibrosis in infected lungs. Using immunohistochemistry, Western blot, and RT-PCR techniques, we demonstrated that recruited alveolar macrophages showed high levels of expression of the proteins Ym1/2, FIZZ1 (found in inflammatory zone 1), insulin-like growth factor-1, and arginase I, and also active transcription of fibronectin, indicative of activation of macrophages by an alternative pathway. Arginase I expression was also evident in interstitial fibroblasts, and increased arginase activity was found in lungs of infected animals. Lung tissue from patients with IPF showed increased expression of arginase I in epithelial cells, fibroblast foci, and alveolar macrophages compared with normal lung. These results suggest that virus-induced upregulation of arginase I could be a mechanism driving lung fibrogenesis. PMID- 16709960 TI - Effects of nitric oxide synthases in chronic allergic airway inflammation and remodeling. AB - The precise role of each nitric oxide (NO) synthase (NOS) isoform in the pathobiology of asthma is not well established. Our objective was to investigate the contribution of constitutive NO synthase (cNOS) and inducible NOS (iNOS) isoforms to lung mechanics and inflammatory and remodeling responses in an experimental model of chronic allergic pulmonary inflammation. Guinea pigs were submitted to seven ovalbumin exposures with increasing doses (1 approximately 5 mg/ml) for 4 wk. The animals received either chronic L-NAME (N-nitro-L-arginine methyl ester, in drinking water) or 1,400 W (iNOS-specific inhibitor, intraperitoneal) treatments. At 72 h after the seventh inhalation of ovalbumin solution, animals were anesthetized, mechanically ventilated, exhaled NO was collected, and lung mechanical responses were evaluated before and after antigen challenge. Both L-NAME and 1,400 W treatments increased baseline resistance and decreased elastance of the respiratory system in nonsensitized animals. After challenge, L-NAME increased resistance of the respiratory system and collagen deposition on airways, and decreased peribronchial edema and mononuclear cell recruitment. Administration of 1,400 W reduced resistance of the respiratory system response, eosinophilic and mononuclear cell recruitment, and collagen and elastic fibers content in airways. L-NAME treatment reduced both iNOS- and neuronal NOS-positive eosinophils, and 1,400 W diminished only the number of eosinophils expressing iNOS. In this experimental model, inhibition of NOS derived NO by L-NAME treatment amplifies bronchoconstriction and increases collagen deposition. However, blockage of only iNOS attenuates bronchoconstriction and inflammatory and remodeling processes. PMID- 16709959 TI - Hypoxia-induced mitogenic factor promotes vascular adhesion molecule-1 expression via the PI-3K/Akt-NF-kappaB signaling pathway. AB - Hypoxia-induced mitogenic factor (HIMF), also known as FIZZ1 (found in inflammatory zone 1), is an important player in lung inflammation. However, the effects of HIMF on cell adhesion molecules involved in lung inflammation remain largely unknown. In the present work, we tested whether HIMF modulates vascular adhesion molecule (VCAM)-1 expression, and dissected the possible signaling pathways that link HIMF to VCAM-1 upregulation. Recombinant HIMF protein, instilled intratracheally into adult mouse lungs, results in a significant increase of VCAM-1 production in vascular endothelial, alveolar type II, and airway epithelial cells. In cultured mouse endothelial SVEC 4-10 and lung epithelial MLE-12 cells, we demonstrated that HIMF induces VCAM-1 expression via the phosphatidylinositol-3 kinase (PI-3K)/Akt-nuclear factor (NF)-kappaB signaling pathway. Knockdown of HIMF expression by small interference RNA attenuated LPS-induced VCAM-1 expression in vitro. We showed that HIMF induced phosphorylation of the IkappaB kinase signalsome and, subsequently, IkappaBalpha, leading to activation of NF-kappaB. Meanwhile, VCAM-1 production was correspondingly upregulated. Blocking NF-kappaB signaling pathway by expression of dominant-negative mutants of IkappaB kinase and IkappaBalpha suppressed HIMF induced VCAM-1 upregulation. HIMF also strongly induced phosphorylation of Akt. A dominant-negative mutant of PI-3K, Deltap85, as well as PI-3K inhibitor, LY294002, also blocked HIMF-induced NF-kappaB activation and attenuated VCAM-1 production. Furthermore, LY294002 pretreatment abolished HIMF-enhanced mononuclear cells adhesion to endothelial and epithelial cells. Our findings connect HIMF to signaling pathways that regulate inflammation, and thus reveal the critical roles that HIMF plays in lung inflammation. PMID- 16709964 TI - Ethnicity and adverse drug reactions. PMID- 16709965 TI - A new human genotype prone to variant Creutzfeldt-Jakob disease. PMID- 16709961 TI - Adenosine A1 receptors mediate mobilization of calcium in human bronchial smooth muscle cells. AB - Adenosine stimulates contraction of airway smooth muscle, but the mechanism is widely considered indirect, depending on release of contractile agonists from mast cells and nerves. The goal was to determine whether adenosine, by itself, directly regulates calcium signaling in human bronchial smooth muscle cells (HBSMC). Primary cultures of HBSMC from normal subjects were loaded with fura 2 AM, and cytosolic calcium concentrations ([Ca(2+)](i)) were determined ratiometrically by imaging single cells. The nonselective adenosine receptor agonist, 5'-N-ethylcarboxamidoadenosine (NECA), and the adenosine A(1) receptor agonist, N(6)-cyclopentyladenosine (CPA), both stimulated rapid, transient increases in [Ca(2+)](i). In contrast, there were no calcium responses to 2-p-(2 carboxyethyl)phenethylamino-5'-N-ethylcarboxamido-adenosine (100 nM) or N(6)-(3 iodobenzyl)-adenosine-5'-N-methyluronamide (100 nM), selective agonists at adenosine A(2A) receptors and adenosine A(3) receptors, respectively. Calcium responses to NECA and CPA were inhibited by 8-cyclopentyl-1,3-dipropylxanthine, an adenosine A(1) receptor antagonist, and by pertussis toxin (PTX). In other experiments, NECA stimulated calcium transients in the absence of extracellular calcium, but not when cells were preincubated in cyclopiazonic acid or thapsigargin to empty intracellular calcium stores. Calcium responses were attenuated by xestospongin C and 2-aminoethoxydiphenylborane, inhibitors of inositol trisphosphate (IP(3)) receptors, and by U73122, an inhibitor of phospholipase C. It was concluded that stimulation of adenosine A(1) receptors on HBSMC rapidly mobilizes intracellular calcium stores by a mechanism dependent on PTX-sensitive G proteins, and IP(3) signaling. These findings suggest that, in addition to its well-established indirect effects on HBSMC, adenosine also has direct effects on contractile signaling pathways. PMID- 16709966 TI - Improved food intake in frail older people. PMID- 16709967 TI - Decent health care for older people. PMID- 16709969 TI - UK House of Lords rejects physician assisted suicide. PMID- 16709971 TI - Charity warns of climate impact on poorer nations. PMID- 16709972 TI - Poor checks for bedside blood transfusion put patients at risk. PMID- 16709973 TI - Aid withdrawal is bringing health service in Gaza to brink of collapse. PMID- 16709974 TI - Can GPs compete with big business? PMID- 16709977 TI - Switzerland launches naked poster campaign to stop AIDS. PMID- 16709979 TI - HFEA widens its criteria for preimplantation genetic diagnosis. PMID- 16709980 TI - Roche denies claims it sacked employee for "whistle blowing". PMID- 16709983 TI - Manufacturer admits increase in suicidal behaviour in patients taking paroxetine. PMID- 16709984 TI - Body consults to balance public health against private rights. PMID- 16709987 TI - Indian pressure group challenges company's right to patent AIDS drug. PMID- 16709988 TI - WHO ultimatum on artemisinin monotherapy is showing results. PMID- 16709989 TI - Tomsk--a glimpse of one Russian region's experience with HIV/AIDS. PMID- 16709990 TI - AIDS in Russia--glasnost arrives at last. PMID- 16709991 TI - Underdosing of antiretrovirals in UK and Irish children with HIV as an example of problems in prescribing medicines to children, 1997-2005: cohort study. AB - OBJECTIVE: To measure the extent of underdosing of antiretroviral drugs in children. DESIGN: Multicentre cohort study. SETTING: Clinical centres in hospitals in the United Kingdom and Ireland in the collaborative HIV paediatric study (CHIPS). PARTICIPANTS: 615 HIV infected children aged 2-12 years receiving antiretrovirals. MAIN OUTCOME MEASURES: Doses relative to weight and height compared with current recommended doses in 2004 European guidelines. RESULTS: The CHIPS cohort of 934 children comprises 80% of diagnosed HIV infected children in the UK and Ireland between January 1997 and March 2005, of which 66% (615) aged 2 12 years were prescribed antiretrovirals. Actual doses standardised to weight or surface area varied widely across individual drugs, antiretroviral class, and calendar time, with children underdosed (prescribed less than 90% of current recommended doses) from 6-62% child time at risk. Three serious issues in prescribing antiretrovirals, which may also be relevant to paediatric prescribing in general, were identified. Firstly, dosing was inadequate before incorrect recommendations at licensing were later revised when important pharmacokinetic results emerged. Secondly, guidelines stating dosage alternatives (by weight/surface area) for the same drug led to different and inconsistent doses. And, thirdly, ongoing growth was not adjusted for. CONCLUSIONS: Largely inadvertently, HIV infected children in the United Kingdom and Ireland have been underdosed with antiretrovirals, highlighting problems applicable throughout paediatric prescribing. PMID- 16709992 TI - Gabapentin may cause reversible visual field constriction. PMID- 16709993 TI - Pulmonary tuberculosis: diagnosis and treatment. PMID- 16709994 TI - 70 year old man with scrotal swelling, abdominal aortic aneurysm, and renal lesions: case outcome. PMID- 16710002 TI - How social marketing works in health care. PMID- 16710004 TI - Putting social marketing into practice. PMID- 16710005 TI - Pneumonia: macrolides or amoxicillin for community acquired pneumonia? PMID- 16710006 TI - Pneumonia: are we putting the CURB score into practice? PMID- 16710007 TI - Peripheral arterial disease: still on the periphery? PMID- 16710008 TI - How should we rate research? Counting number of publications may be best research performance measure. PMID- 16710009 TI - Pneumonia: let's avoid confusion of secondary and primary care issues in pneumonia. PMID- 16710010 TI - Pneumonia: ... and to tuberculosis as differential diagnosis in community acquired pneumonia. PMID- 16710011 TI - Pneumonia: be alert to underlying malignancy in follow up of pneumonia... PMID- 16710012 TI - Self harm in Goth youth subculture: study merely reinforces popular stereotypes. PMID- 16710013 TI - How should we rate research? Pedigree: paternity and progeny. PMID- 16710014 TI - How should we rate research? Cost of research should be disclosed publicly. PMID- 16710015 TI - Time for evidence based rewards? PMID- 16710016 TI - New term may be needed for data categorisation. PMID- 16710017 TI - Rectal quinine for malaria: risk of hypoglycaemia may be higher by rectal route. PMID- 16710018 TI - Self harm in Goth youth subculture: conclusion relates only to small sample. PMID- 16710019 TI - Is there a role for the irreversible epidermal growth factor receptor inhibitor EKB-569 in the treatment of cancer? A mutation-driven question. PMID- 16710020 TI - Is tumor immunity the same thing as autoimmunity? Implications for cancer immunotherapy. PMID- 16710021 TI - Epoetin alfa and darbepoetin alfa go head to head. PMID- 16710022 TI - Never-smokers with lung cancer: epidemiologic evidence of a distinct disease entity. AB - PURPOSE: Tobacco smoke is a definite causative agent for lung cancer. It is increasingly being recognized that never-smokers can be afflicted with non-small cell lung cancer (NSCLC). We aim to assess survival differences between smokers and never-smokers with NSCLC. PATIENTS AND METHODS: We analyzed 975 NSCLC patients who presented from January 1999 to December 2002. Clinical characteristics among current-, former- and never-smokers were tested using chi2 or Kruskal-Wallis test. The hazard ratio (HR) for death and its 95% CI were calculated by Cox regression. RESULTS: Of 975 patients, 59 had no smoking history and 33 had no quit time recorded. Of 883 patients analyzed, 286 patients (32.4%) were never-smokers. One hundred ninety-six never-smokers (68.5%) were females compared with 12% among current- and 13% among former-smokers (P < .001). There was a significant difference in histologic subtype between never-smokers and smokers: 69.9% with adenocarcinoma versus 39.9% (current-smokers) versus 47.3% (former-smokers); 5.9% with squamous cell carcinoma versus 35.7% (current smokers) versus 28% (former-smokers; P < .001). Smokers had significantly poorer performance status (P = .002) and higher median age at diagnosis (P < .001) while more never-smokers presented with advanced disease (P = .002). Eight hundred and five patients (82.6%) died by May 30, 2005. The HR for smokers was significantly higher on both univariate and multivariate analysis (HR, 1.297; 95% CI, 1.040 to 1.619). CONCLUSION: Never-smokers comprised a high proportion of NSCLC patients in Singapore. Definite epidemiologic differences exist between never-smokers and smokers. Differences in survival outcome further suggest that the biology underlying the pathogenesis and behavior of the disease may be different for never-smokers. PMID- 16710023 TI - Phase I study of EKB-569, an irreversible inhibitor of the epidermal growth factor receptor, in patients with advanced solid tumors. AB - PURPOSE: The maximum tolerated dose (MTD) and the dose-limiting toxicities of EKB 569, a selective, irreversible inhibitor of the epidermal growth factor receptor (EGFR), when administered orally once daily on an intermittent-dose schedule (14 days of a 28-day cycle) or on a continuous-dose schedule (each day of a 28-day cycle), were determined in patients with advanced solid tumors. PATIENTS AND METHODS: Planned dose escalation was 25, 50, 75, 125, 175, and 225 mg. Pharmacokinetic sampling was performed on days 1 and 14 for the intermittent-dose cohort and on days 1 and 15 for the continuous-dose cohort. RESULTS: Thirty patients received a median of two cycles (range, one to 10 cycles) in the intermittent-dose cohort; 29 patients received a median of three cycles (range, one to eight cycles) in the continuous-dose cohort. Dose-limiting toxicity was grade 3 diarrhea, and the MTD was 75 mg EKB-569 per day for both cohorts. Other common toxicities included rash, nausea, and asthenia. Exposure to EKB-569 was dose proportional. At the MTD, the mean +/- standard deviation terminal half-life was 21.7 +/- 4.2 hours and peak concentration increased 1.2-fold from day 1 to day 14/15. No major antitumor responses were observed. However, one patient with non-small-cell lung cancer and one with cutaneous squamous cell carcinoma had stable disease for 33 and 24 weeks, respectively. CONCLUSION: The MTD of once daily oral EKB-569 is 75 mg. The tolerable toxicity profile and long half-life of this irreversible EGFR inhibitor warrant its further evaluation as a single agent and in combination with other drugs. PMID- 16710024 TI - Indium-111-labeled trastuzumab scintigraphy in patients with human epidermal growth factor receptor 2-positive metastatic breast cancer. AB - PURPOSE: The cardiac and antineoplastic effects of trastuzumab may be related to specific uptake of trastuzumab in myocardium and tumor tissue, respectively. We evaluated whether indium-111 (111In)-labeled trastuzumab scintigraphy can predict cardiotoxicity and identify tumor lesions. In addition, we evaluated whether plasma markers for cardiac dysfunction can be used to predict cardiotoxicity. PATIENTS AND METHODS: Patients with human epidermal growth factor receptor 2 (HER2) -positive metastatic breast cancer underwent gamma camera imaging from 15 minutes to 7 days after injection of 150 MBq 111In-diethylenetriamine penta acetic acid anhydride (DTPA) -trastuzumab, after loading-dose trastuzumab, and after once-a-week trastuzumab doses for 11 weeks, and concomitant paclitaxel once every 3 weeks. Cardiac assessments were performed before treatment, and after four and six cycles. Plasma N-terminal probrain natriuretic peptide (NT-proBNP) and serum troponin I were measured with immunoassay. RESULTS: Fifteen of the 17 patients were available for cardiac and tumor uptake analysis. On the first scan, myocardial 111In-DTPA-trastuzumab uptake was observed in one patient with pre existing cardiac arrhythmias, who did not develop heart failure during treatment. Severe cardiotoxicity occurred in three patients, without initial myocardial uptake, whereas one showed weak myocardial uptake after four cycles. The detection rate of single tumor lesions was 45%. New tumor lesions were discovered in 13 of 15 patients. Pretreatment plasma NT-proBNP levels were higher in patients with than without heart failure (mean, 534 [standard deviation, 236] v 105 [standard deviation, 79] ng/L; P = .009). CONCLUSION: Radiolabeled trastuzumab scintigraphy was not valuable in predicting trastuzumab-related cardiotoxicity in metastatic breast cancer patients, but can identify HER2 positive tumors. Measurement of plasma NT-proBNP is promising regarding prediction of trastuzumab-related cardiotoxicity. PMID- 16710026 TI - Randomized comparison of every-2-week darbepoetin alfa and weekly epoetin alfa for the treatment of chemotherapy-induced anemia: the 20030125 Study Group Trial. AB - PURPOSE: Chemotherapy-induced anemia is widely treated in the United States with darbepoetin alfa (DA) or epoetin alfa (EA). This noninferiority study systematically compares efficacy and safety of DA and EA using common doses and schedules used in clinical practice. METHODS: Patients had a diagnosis of nonmyeloid malignancy with > or = 8 weeks of planned chemotherapy, age >or = 18 years, and anemia (hemoglobin < or = 11 g/dL). Patients were randomly assigned 1:1 to DA 200 microg every two weeks (Q2W) or EA 40,000 units every week (QW) for up to 16 weeks with identical dose adjustment rules. Efficacy was assessed by the incidence of RBC transfusion (Kaplan-Meier estimate). The definition of noninferiority was that the upper 95% CI limit of the observed difference in RBC transfusions between groups was less than 11.5%; this noninferiority margin was based on the treatment effect observed in placebo-controlled EA studies. RESULTS: Of 1,220 patients randomly assigned, 1,209 received > or = one dose of the study drug. Common tumor types were lung (26%), breast (21%), and gastrointestinal (18%). Transfusion incidence from week 5 to the end of the treatment phase (the primary end point) was 21% in the DA group and 16% in the EA group; noninferiority was concluded because the upper 95% CI limit of the difference between groups (10.8%) was below the prespecified noninferiority margin. Sensitivity analyses using alternate statistical methods and analysis sets yielded similar results. Hemoglobin, quality of life, and safety end points further support equivalency of the erythropoietic therapies. CONCLUSION: This large, phase III study demonstrates comparable efficacy of DA Q2W and EA QW. Less frequent dosing offers potential benefits for patients, caregivers and health care providers. PMID- 16710025 TI - Enterocolitis in patients with cancer after antibody blockade of cytotoxic T lymphocyte-associated antigen 4. AB - PURPOSE: Cytotoxic T-lymphocyte-associated antigen 4 (CTLA4) is an inhibitory receptor on T cells. Knocking out CTLA4 in mice causes lethal lymphoproliferation, and polymorphisms in human CTLA4 are associated with autoimmune disease. Trials of the anti-CTLA4 antibody ipilimumab (MDX-010) have resulted in durable cancer regression and immune-mediated toxicities. A report on the diagnosis, pathology, treatment, clinical outcome, and significance of the immune-mediated enterocolitis seen with ipilimumab is presented. PATIENTS AND METHODS: We treated 198 patients with metastatic melanoma (MM) or renal cell carcinoma (RCC) with ipilimumab. RESULTS: The overall objective tumor response rate was 14%. We observed several immune mediated toxicities including dermatitis, enterocolitis, hypophysitis, uveitis, hepatitis, and nephritis. Enterocolitis, defined by grade 3/4 clinical presentation and/or biopsy documentation, was the most common major toxicity (21% of patients). It presented with diarrhea, and biopsies showed both neutrophilic and lymphocytic inflammation. Most patients who developed enterocolitis responded to high-dose systemic corticosteroids. There was no evidence that steroid administration affected tumor responses. Five patients developed perforation or required colectomy. Four other patients with steroid-refractory enterocolitis appeared to respond promptly to tumor necrosis factor alpha blockade with infliximab. Objective tumor response rates in patients with enterocolitis were 36% for MM and 35% for RCC, compared with 11% and 2% in patients without enterocolitis, respectively (P = .0065 for MM and P = .0016 for RCC). CONCLUSION: CTLA4 seems to be a significant component of tolerance to tumor and in protection against immune mediated enterocolitis and these phenomena are significantly associated in cancer patients. PMID- 16710027 TI - Geriatric syndromes in elderly patients admitted to an oncology-acute care for elders unit. AB - PURPOSE: The goal of this study was to characterize an elderly population admitted to a novel Oncology-Acute Care for Elders (OACE) unit, determine the prevalence of functional dependencies and geriatric syndromes, and examine their suitability for an interdisciplinary model of care. PATIENTS AND METHODS: We conducted a retrospective review of 119 patients age 65 years or older who had a primary oncologic or hematologic diagnosis and were admitted to the OACE Unit. Standard geriatric screens were administered to assess mood, functional, and cognitive status. Demographic and medical data were compiled by review of patients' medical records. RESULTS: The mean age of the patients was 74.1 years (standard deviation, 5.9 years). The sample was predominantly white, of equal sex, had limitations in instrumental and basic activities of daily living, and a mean length of stay of 6 days. Geriatric syndromes detected by the OACE interdisciplinary team included cognitive impairment (dementia and/or delirium), depression, weight loss, and use of high-risk medications. Adverse events such as falls, restraint use, and pressure sores were rare. CONCLUSION: In this descriptive study, many older cancer patients were found to have geriatric syndromes by the OACE team and these patients were considered appropriate for an interdisciplinary model of care. Additional studies are needed to compare the outcomes of hospitalized older oncology patients receiving an OACE intervention with those patients receiving usual care. PMID- 16710028 TI - Comorbidity, disability, and geriatric syndromes in elderly cancer patients receiving home health care. AB - PURPOSE: To assess the prevalence of comorbidity, disability, and geriatric syndromes, or a combination thereof, in elders with cancer receiving home health care (HHC). PATIENTS AND METHODS: Using the Ohio Cancer Incidence Surveillance System, we identified Ohio residents 65 years of age or older who were diagnosed with incident breast (n = 952), prostate (n = 324), or colorectal cancer (n = 1,276) during the 28-month study period, August 1999 through November 2001. We used the Outcome and Assessment Information Set, a database compiling comprehensive assessment forms completed for all HHC patients, to group individuals in independent and overlapping categories of comorbidity, disability, and geriatric syndromes on the basis of the patients' clinical condition 14 days before the date of the assessment. RESULTS: The proportion with no comorbidity, disability, or geriatric syndromes was 26.4% in breast cancer patients, 12.0% in prostate cancer patients, and 14.0% in colorectal cancer patients. The proportion of patients presenting all three entities at once was 11.7%, 24.7%, and 15.7%, respectively, in three cancer sites. As expected, the proportion of patients with no comorbidity, disability, or geriatric syndromes declined gradually with increasing age, and that of patients with all three entities was highest among patients 85 years or older. CONCLUSION: The proposed taxonomy will help us gain a more nuanced understanding of older cancer patients' clinical presentation and may lead to a more accurate identification of older patients who might benefit from standard cancer treatment, and those who might experience adverse outcomes. PMID- 16710029 TI - Physical and emotional health effects and social consequences after participation in a low-fat, high-carbohydrate dietary trial for more than 5 years. AB - PURPOSE: Little is known about the potential adverse effects of interventions to reduce dietary fat. We examined the physical and emotional health effects, and social consequences experienced by women at high risk for breast cancer who had participated in a low-fat diet intervention, randomized, controlled trial for at least 5 years. METHODS: Participants in the Canadian Diet and Breast Cancer Prevention Trial from British Columbia were mailed a survey questionnaire that included the validated Medical Outcomes Study 36-item Short Form Health Survey (SF-36) and Women's Health Questionnaire (WHQ), and a series of questions on health-related and social constructs. Responses were compared between the diet intervention and control groups by menopausal status. RESULTS: Completed questionnaires were returned by 359 women in the diet intervention group and 382 in the control group. No significant differences were found between these groups for SF-36 and WHQ health outcomes, hair/nail changes, physical activity levels, family/friend support levels, and doctor visits. Significantly more women in the intervention group reported taking products for arthritis (other than pain medication), greater difficulty in maintaining eating habits in social situations and at work, greater stress, and guilt related to personal eating habits. These findings persisted for both premenopausal and postmenopausal women. CONCLUSION: Changes resulting from a low-fat diet intervention can be incorporated into women's daily lives with limited long-term negative effects. PMID- 16710030 TI - Radiochemotherapy after transurethral resection for high-risk T1 bladder cancer: an alternative to intravesical therapy or early cystectomy? AB - PURPOSE: For high-risk T1 bladder cancer, the most important issue is how to restrict radical cystectomy to selective patients with a high likelihood of tumor progression and to choose an initial bladder-sparing approach in others without affecting survival. Radiotherapy or radiochemotherapy (RT/RCT) may help to strike a balance between intravesical treatment and early cystectomy. PATIENTS AND METHODS: Between 1982 and 2004, 141 patients with high-risk T1 bladder cancer (84 patients with T1 grade 3 [T1G3]; others with T1G1/2 and associated carcinoma-in situ, multifocality, tumor diameter > 5 cm, or multiple recurrences) were treated with RT (n = 28) or platinum-based RCT (n = 113) after transurethral resection of bladder tumor (TURBT). Six weeks after RT/RCT, response was evaluated by restaging TURBT. Salvage cystectomy was recommended for patients with persistent disease and for tumor progression after initial complete response (CR). Median follow-up was 62 months; 65 patients have been observed for 5 years or more. RESULTS: CR was achieved in 121 of 137 patients (88%; four patients without restaging TURBT). Tumor progression for the entire group of 141 patients was 19% and 30% at 5 and 10 years, respectively (for 121 patients with CR, 15% and 29%; for 84 patients with T1G3, 13% and 29%, respectively). Disease-specific survival rates were 82% and 73% at 5 and 10 years (CR, 89% and 79%; T1G3, 80% and 71%, respectively). More than 80% of survivors preserved their bladder; 70.4% were "delighted" or "pleased" with their urinary function. CONCLUSION: RT/RCT after TURBT with selective bladder preservation is a reasonable alternative to intravesical treatment or early cystectomy for high-risk T1 bladder cancer. PMID- 16710031 TI - Surgical management of advanced gastrointestinal stromal tumors after treatment with targeted systemic therapy using kinase inhibitors. AB - PURPOSE: While targeted inhibitors of tyrosine kinase activity demonstrate dramatic efficacy in the majority of patients with advanced gastrointestinal stromal tumors (GISTs), cure remains elusive and resistance to systemic therapy is a challenge. To assess the role of surgery in multimodality management of GISTs, we studied postoperative outcomes in patients treated with targeted kinase inhibitors for advanced GIST. METHODS: We evaluated outcomes in a single institution series of 69 consecutive patients who underwent surgery for advanced GISTs while receiving kinase inhibitors. Patients were categorized based on extent of disease before surgery (stable disease, limited disease progression, generalized disease progression) and surgical result (no evidence of disease, minimal residual disease, bulky residual disease). RESULTS: Disease status before surgery was associated with surgical result (P < .0001; median follow-up, 14.6 months). After surgery, there was no evidence of disease in 78%, 25%, and 7% of patients with stable disease, limited progression, and generalized progression, respectively. Bulky residual disease remained after surgery in 4%, 16%, and 43% of the patients with stable disease, limited progression, and generalized progression. Twelve-month progression-free survival was 80%, 33%, and 0% for patients with stable disease, limited progression, and generalized progression (P < .0001). Twelve-month overall survival was 95%, 86%, and 0% for patients with stable disease, limited progression, and generalized progression (P < .0001). CONCLUSION: Patients with advanced GISTs exhibiting stable disease or limited progression on kinase inhibitor therapy have prolonged overall survival after debulking procedures. Surgery has little to offer in the setting of generalized progression. PMID- 16710032 TI - Prognostic significance of blasts in the cerebrospinal fluid without pleiocytosis or a traumatic lumbar puncture in children with acute lymphoblastic leukemia: experience of the Dutch Childhood Oncology Group. AB - PURPOSE: To determine the significance of blasts in the CSF without pleiocytosis and a traumatic lumbar puncture in children with acute lymphoblastic leukemia (ALL). PATIENTS AND METHODS: We retrospectively studied a cohort of 526 patients treated in accordance with the virtually identical Dutch protocols ALL-7 and ALL 8. Patients were classified into five groups: CNS1, no blasts in the CSF cytospin; CNS2, blasts present in the cytospin, but leukocytes less than 5/microL; CNS3, blasts present and leukocytes more than 5/microL. Patients with a traumatic lumbar puncture (TLP; > 10 erythrocytes/mL) were classified as TLP+ (blasts present in the cytospin) or TLP- (no blasts). RESULTS: Median duration of follow-up was 13.2 years (range, 6.9 to 15.5 years). Event-free survival (EFS) was 72.6% (SE, 2.5%) for CNS1 patients (n = 304), 70.3% (SE, 4.7%) for CNS2 patients (n = 111), and 66.7% (SE, 19%) for CNS3 patients (n = 10; no significant difference in EFS between the groups). EFS was 58% (SE, 7.6%) for TLP+ patients (n = 62) and 82% (SE, 5.2%) for TLP- patients (n = 39; P < .01). Cox regression analysis identified TLP+ status as an independent prognostic factor (risk ratio, 3.5; 95% CI, 1.4 to 8.8; P = .007). Cumulative incidence of CNS relapses was 0.05 and 0.07 in CNS1 and CNS2 patients, respectively (not statistically significant). CONCLUSION: In our experience, the presence of a low number of blasts in the CSF without pleiocytosis has no prognostic significance. In contrast, a traumatic lumbar puncture with blasts in the CSF specimen is associated with an inferior outcome. PMID- 16710033 TI - Clinical outcomes and prognostic factors in patients with Richter's syndrome treated with chemotherapy or chemoimmunotherapy with or without stem-cell transplantation. AB - PURPOSE: The purpose of this study was to assess the incidence, presenting characteristics, and treatment outcomes of Richter's syndrome (RS) and factors predicting response and survival. PATIENTS AND METHODS: An electronic database search of patients with chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma (SLL) who presented at The University of Texas M.D. Anderson Cancer Center (Houston, TX) between January 1975 and June 2005 was performed, and patient medical records were reviewed. RESULTS: Of the 3,986 patients with CLL/SLL, 204 patients (5.1%) had possible RS, and 148 patients (3.7%) had biopsy- or fine-needle aspiration-proven RS. Treatment included chemotherapy alone and chemoimmunotherapy with rituximab. The overall response rate for the 130 assessable patients was 39% (chemotherapy, 34%; chemoimmunotherapy, 47%; P = .2). In multivariate analysis, factors predicting prolonged survival were Zubrod performance status 0-1 (P = .006), lactate dehydrogenase < or = 1.5x the upper limit of normal (P = .003), platelet count > or = 100,000 (P = .01), tumor size < or = 5 cm (P = .02), and fewer than two prior therapies (P = .02). The five adverse factors predicting shorter survival were used to design a model to predict an individual patient's risk of death: the RS score. A total of 20 patients underwent stem-cell transplantation (SCT). Patients who underwent allogeneic SCT as postremission therapy had longer survival than patients who achieved remission and received no additional therapy or patients who underwent allogeneic or autologous SCT as salvage therapy (P = .019). CONCLUSION: A score to predict an individual patient's risk of death is proposed. Chemotherapy and rituximab combinations are effective in RS. Patients with available donors may be considered for allogeneic SCT as postremission therapy. PMID- 16710034 TI - Treatment of anaplastic histology Wilms' tumor: results from the fifth National Wilms' Tumor Study. AB - PURPOSE: An objective of the fifth National Wilms' Tumor Study (NWTS-5) was to evaluate the efficacy of treatment regimens for anaplastic histology Wilms' tumor (AH). PATIENTS AND METHODS: Prospective single-arm studies were conducted. Patients with stage I AH were treated with vincristine and dactinomycin for 18 weeks. Patients with stages II to IV diffuse AH were treated with vincristine, doxorubicin, cyclophosphamide, and etoposide for 24 weeks plus flank/abdominal radiation. RESULTS: A total of 2,596 patients with Wilms' tumor were enrolled onto NWTS-5, of whom 281 (10.8%) had AH. Four-year event-free survival (EFS) and overall survival (OS) estimates for assessable patients with stage I AH (n = 29) were 69.5% (95% CI, 46.9 to 84.0) and 82.6% (95% CI, 63.1 to 92.4). In comparison, 4-year EFS and OS estimates for patients with stage I favorable histology (FH; n = 473) were 92.4% (95% CI, 89.5 to 94.5) and 98.3% (95% CI, 96.4 to 99.2). Four-year EFS estimates for patients who underwent immediate nephrectomy with stages II (n = 23), III (n = 43), and IV (n = 15) diffuse AH were 82.6% (95% CI, 60.1 to 93.1), 64.7% (95% CI, 48.3 to 77.7), and 33.3% (95% CI, 12.2 to 56.4), respectively. OS was similar to EFS for these groups. There were no local recurrences among patients with stage II AH. Four-year EFS and OS estimates for patients with bilateral AH (n = 29) were 43.8% (95% CI, 24.2 to 61.8) and 55.2% (95% CI, 34.8 to 71.7), respectively. CONCLUSION: The prognosis for patients with stage I AH is worse than that for patients with stage I FH. Novel treatment strategies are needed to improve outcomes for patients with AH, especially those with stage III to V disease. PMID- 16710035 TI - Immunohistochemical test for MLH1 and MSH2 expression predicts clinical outcome in stage II and III colorectal cancer patients. AB - PURPOSE: To evaluate the prognostic significance of DNA mismatch repair (MMR) status in a large series of stage II and III colorectal cancer patients. The relationship among MMR status, adjuvant chemotherapy, and clinical outcome was also investigated. PATIENTS AND METHODS: The study included 718 patients with colorectal adenocarcinoma (393 stage II and 325 stage III) who underwent curative surgical resection. MMR status was determined by immunohistochemical analysis of MLH1 and MSH2 expression. Microsatellite instability (MSI) was assessed in 363 patients using mononucleotide and dinucleotide markers. RESULTS: One hundred fourteen (15.9%) carcinomas showed abnormal MMR protein (MMRP) expression (96 MLH1 negative and 18 MSH2 negative) and were classified as MMRP negative, whereas 604 tumors demonstrated normal MLH1/MSH2 immunoreactivity (MMRP positive). MLH1/MSH2 expression was closely related to MSI status (P < .001) and several clinicopathologic features. Patients with MMRP-negative carcinomas demonstrated a marked reduction in the risk of cancer-related death with respect to patients with MMRP-positive tumors (hazard ratio, 0.2579; 95% CI, 0.1289 to 0.5159). A better clinical outcome for patients with MMRP-negative tumors was observed in both stage II (P = .0006) and stage III (P = .0052) disease. In stage III disease, the survival advantage conferred by MMRP-negative tumors was more evident among patients treated with surgery alone than among patients who received adjuvant chemotherapy. A nonsignificant trend for survival benefit from adjuvant chemotherapy was observed among patients with MMRP-positive carcinomas but not among those with MMRP-negative carcinomas. CONCLUSION: Immunohistochemical testing for MLH1/MSH2 expression provides useful prognostic information for the management of stage II and III colorectal cancer patients. PMID- 16710036 TI - HER-2 is an independent prognostic factor in endometrial cancer: association with outcome in a large cohort of surgically staged patients. AB - PURPOSE: To evaluate HER-2 expression and amplification in a large cohort of endometrial cancer with complete surgical staging and outcome data. PATIENTS AND METHODS: A tissue microarray was constructed of 483 patients with endometrial cancer of diverse histologic type and stage and tested for HER-2 expression and amplification using current standards of practice. There was outcome data for 83% of all patients and 81% with complete surgical staging. RESULTS: Both expression and amplification of HER-2 was associated with high-grade (P = .0001) and high stage (P = .0001) endometrial cancer. The highest rate of HER-2 expression and amplification was seen in serous carcinoma (43% and 29%), while grade 1 endometrioid adenocarcinoma showed the lowest levels (3% and 1%). For all histologic types, the rate of HER-2 expression and amplification was remarkably different (P < .0001) for grade 3 cancers (31% and 15%) versus grade 2 (7% and 3%) and grade 1 cancers (3% and 1%), with similar results for endometrioid type (P < .0001). Both HER-2 expression and amplification correlated with disease specific survival and progression-free survival in univariate analyses. By multivariate analysis HER-2 expression in the presence of amplification (P = .012) correlated with overall survival, but not expression in the absence of amplification. Overall survival was significantly shorter (P = .0001) in patients who overexpressed (median, 5.2 years) and/or showed amplification of HER-2 (median, 3.5 years) versus those that did not (median of all cases, 13 years). CONCLUSION: Our results would suggest that HER-2 is an important oncogene in high grade and stage endometrial cancer, but plays only a minor role in the much more common low grade and stage tumors that encompass the majority of clinical practice. PMID- 16710038 TI - Intravascular hemolysis as a complication of clostridium perfringens sepsis. PMID- 16710037 TI - Metachronous intracranial germinoma in a patient with a previous primary mediastinal seminoma. PMID- 16710039 TI - Paraneoplastic erythropoietin-induced polycythemia associated with small lymphocytic lymphoma. PMID- 16710040 TI - Tumor lysis syndrome after treatment with docetaxel for non-small-cell lung cancer. PMID- 16710041 TI - Myeloid toxicity in breast cancer patients receiving adjuvant chemotherapy with pegfilgrastim support. PMID- 16710042 TI - Effect of tamoxifen after chemotherapy in hormone receptor-positive, node negative breast cancer. PMID- 16710043 TI - Aggressive surgery and ovarian cancer. PMID- 16710044 TI - Importance of surgical aggressiveness in advanced ovarian cancer. PMID- 16710045 TI - Acute chemotherapy-induced cardiovascular changes in patients with testicular cancer: are there implications for blood pressure management in patients receiving chemotherapy? PMID- 16710046 TI - Expressions of concern. PMID- 16710048 TI - The scarlet letters. PMID- 16710047 TI - Absence of VLDL secretion does not affect alpha-tocopherol content in peripheral tissues. AB - alpha-Tocopherol is a lipid-soluble antioxidant that helps to prevent oxidative damage to cellular lipids. alpha-Tocopherol is absorbed by the intestine and is taken up and retained by the liver; it is widely presumed that alpha-tocopherol is then delivered to peripheral tissues by the secretion of VLDL. To determine whether VLDL secretion is truly important for the delivery of alpha-tocopherol to peripheral tissues, we examined alpha-tocopherol metabolism in mice that lack microsomal triglyceride transfer protein (Mttp) expression in the liver and therefore cannot secrete VLDL (Mttp(Delta/Delta) mice). Mttp(Delta/Delta) mice have low plasma lipid levels and increased stores of lipids in the liver. Similarly, alpha-tocopherol levels in the plasma were lower in Mttp(Delta/Delta) mice than in controls, whereas hepatic alpha-tocopherol stores were higher. However, alpha-tocopherol levels in the peripheral tissues of Mttp(Delta/Delta) mice were nearly identical to those of control mice, suggesting that VLDL secretion is not critical for the delivery of alpha-tocopherol to peripheral tissues. When fed a diet containing deuterated alpha-tocopherol, Mttp(Delta/Delta) and control mice had similar incorporation of deuterated alpha tocopherol into plasma and various peripheral tissues. We conclude that the absence of VLDL secretion has little effect on the stores of alpha-tocopherol in peripheral tissues, at least in the mouse. PMID- 16710049 TI - Osteochondral autograft transplantation in the porcine knee. AB - BACKGROUND: Knee articular cartilage defects are not an uncommon problem. Because articular cartilage is limited in its ability to heal, these defects are difficult to manage. HYPOTHESIS: Osteochondral autografts will provide less of a cavitary defect and more viable hyaline articular cartilage than will control knees. STUDY DESIGN: Controlled laboratory study. METHODS: Osteochondral autografts were grossly and microscopically evaluated in the porcine knee and compared with a control at 6 weeks, 3 months, and 6 months. In 18 porcine specimens, a 1-stage surgical procedure was performed to harvest an osteochondral graft from a nonweightbearing articular cartilage surface, and the graft was transplanted into a defect created in the weight-bearing region of the medial femoral condyle. In the opposite control knee, a similar defect was created in the medial femoral condyle; an osteochondral transplant was not performed. Six pigs each were sacrificed at 6 weeks, 3 months, and 6 months. RESULTS: Gross inspection of the control knees showed a cavitary defect. The defect grossly decreased in size with fibrous ingrowth seen on microscopic analysis. An increasing amount of fibrous tissue and fibrocartilage was present at the 3 time periods. Gross inspection of the graft knee showed a healed osteochondral plug with no obvious displacement, cavitary defects, or surrounding necrotic tissue at each time interval. Microscopic analysis revealed the graft knee contained viable hyaline cartilage and healed viable subchondral bone. At all time intervals, 75% to 100% of the hyaline cartilage was viable in all specimens. In 6-month specimens, bridging cartilage at the autograft-host junction was incomplete in 50%, partial in 33%, and complete in 17%. CONCLUSION: Osteochondral autografts in the porcine knee resulted in viable hyaline cartilage for up to 6 months; there was inconsistent bridging hyaline cartilage at the periphery. Grafts appeared to heal into existing subchondral bone without displacement or evidence of necrosis. CLINICAL RELEVANCE: This type of osteochondral transplant can be used as a reliable reconstructive alternative for osteochondral defects. PMID- 16710050 TI - Bile acids stimulate PKCalpha autophosphorylation and activation: role in the attenuation of prostaglandin E1-induced cAMP production in human dermal fibroblasts. AB - The aim was to identify the specific PKC isoform(s) and their mechanism of activation responsible for the modulation of cAMP production by bile acids in human dermal fibroblasts. Stimulation of fibroblasts with 25-100 microM of chenodeoxycholic acid (CDCA) and ursodeoxycholic acid (UDCA) led to YFP-PKCalpha and YFP-PKCdelta translocation in 30-60 min followed by a transient 24- to 48-h downregulation of the total PKCalpha, PKCdelta, and PKCepsilon protein expression by 30-50%, without affecting that of PKCzeta. Increased plasma membrane translocation of PKCalpha was associated with an increased PKCalpha phosphorylation, whereas increased PKCdelta translocation to the perinuclear domain was associated with an increased accumulation of phospho-PKCdelta Thr505 and Tyr311 in the nucleus. The PKCalpha specificity on the attenuation of cAMP production by CDCA was demonstrated with PKC downregulation or inhibition, as well as PKC isoform dominant-negative mutants. Under these same conditions, neither phosphatidylinositol 3-kinase, p38 MAP kinase, p42/44 MAP kinase, nor PKA inhibitors had any significant effect on the CDCA-induced cAMP production attenuation. CDCA concentrations as low as 10 microM stimulated PKCalpha autophosphorylation in vitro. This bile acid effect required phosphatidylserine and was completely abolished by the presence of Go6976. CDCA at concentrations less than 50 microM enhanced the PKCalpha activation induced by PMA, whereas greater CDCA concentrations reduced the PMA-induced PKCalpha activation. CDCA alone did not affect PKCalpha activity in vitro. In conclusion, although CDCA and UDCA activate different PKC isoforms, PKCalpha plays a major role in the bile acid-induced inhibition of cAMP synthesis in fibroblasts. This study emphasizes potential consequences of increased systemic bile acid concentrations and cellular bile acid accumulation in extrahepatic tissues during cholestatic liver diseases. PMID- 16710051 TI - Role of mTOR signaling in intestinal cell migration. AB - An early signaling event activated by amino acids and growth factors in many cell types is the phosphorylation of the mammalian target of rapamycin (mTOR; FRAP), which is functionally linked to ribosomal protein s6 kinase (p70(s6k)), a kinase that plays a critical regulatory role in the translation of mRNAs and protein synthesis. We previously showed that intestinal cell migration, the initial event in epithelial restitution, is enhanced by l-arginine (ARG). In this study, we used amino acids as prototypic activators of mTOR and ARG, IGF-1, or serum as recognized stimulators of intestinal cell migration. We found that 1) protein synthesis is required for intestinal cell migration, 2) mTOR/p70(s6k) pathway inhibitors (rapamycin, wortmannin, and intracellular Ca(2+) chelation) inhibit cell migration, 3) ARG activates migration and mTOR/p70(s6k) (but not ERK-2) in migrating enterocytes, and 4) immunocytochemistry reveals abundant p70(s6k) staining in cytoplasm, whereas phospho-p70(s6k) is virtually all intranuclear in resting cells but redistributes to the periphery on activation by ARG. We conclude that mTOR/p70(s6k) signaling is essential to intestinal cell migration, is activated by ARG, involves both nuclear and cytoplasmic events, and may play a role in intestinal repair. PMID- 16710052 TI - Evidence for epithelial-mesenchymal transitions in adult liver cells. AB - Both myofibroblastic hepatic stellate cells (HSC) and hepatic epithelial progenitors accumulate in damaged livers. In some injured organs, the ability to distinguish between fibroblastic and epithelial cells is sometimes difficult because cells undergo epithelial-mesenchymal transitions (EMT). During EMT, cells coexpress epithelial and mesenchymal cell markers. To determine whether EMT occurs in adult liver cells, we analyzed the expression profile of primary HSC, two HSC lines, and hepatic epithelial progenitors. As expected, all HSC expressed HSC markers. Surprisingly, these markers were also expressed by epithelial progenitors. In addition, one HSC line expressed typical epithelial progenitor mRNAs, and these epithelial markers were inducible in the second HSC line. In normal and damaged livers, small ductular-type cells stained positive for an HSC marker. In conclusion, HSC and hepatic epithelial progenitors both coexpress epithelial and mesenchymal markers, providing evidence that EMT occurs in adult liver cells. PMID- 16710053 TI - Taste receptors in the gastrointestinal tract. I. Bitter taste receptors and alpha-gustducin in the mammalian gut. AB - Molecular sensing by gastrointestinal (GI) cells plays a critical role in the control of multiple fundamental functions in digestion and also initiates hormonal and/or neural pathways leading to the regulation of caloric intake, pancreatic insulin secretion, and metabolism. Molecular sensing in the GI tract is also responsible for the detection of ingested harmful drugs and toxins, thereby initiating responses critical for survival. The initial recognition events and mechanism(s) involved remain incompletely understood. The notion to be discussed in this article is that there are important similarities between the chemosensory machinery elucidated in specialized neuroepithelial taste receptor cells of the lingual epithelium and the molecular transducers localized recently in enteroendocrine open GI cells that sense the chemical composition of the luminal contents of the gut. PMID- 16710054 TI - Distinct mechanisms of acid-induced HCO3- secretion in normal and slightly permeable stomachs. AB - We investigated the regulatory mechanism of acid-induced HCO(3)(-) secretion in the slightly permeable rat stomach after an exposure to hyperosmolar NaCl. Under urethane anesthesia, a rat stomach was mounted on a chamber and perfused with saline, and the secretion of HCO(3)(-) was measured at pH 7.0 using a pH-stat method and by adding 2 mM HCl. Acidification of the normal stomach with 100 mM HCl increased HCO(3)(-) secretion, and this response was totally inhibited by pretreatment with indomethacin but not N(G)-nitro-l-arginine methyl ester (l NAME) or chemical ablation of capsaicin-sensitive afferent neurons. Exposure of the stomach to 0.5 M NaCl deranged the unstirred mucus gel layer without damaging the surface epithelial cells. The stomach responded to 0.5 M NaCl by secreting slightly more HCO(3)(-), in an indomethacin-inhibitable manner, and responded to even 10 mM HCl with a marked rise in HCO(3)(-) secretion, although 10 mM HCl did not have an effect in the normal stomach. The acid-induced HCO(3)(-) response in the NaCl-treated stomach was significantly but partially attenuated by indomethacin, l-NAME, or sensory deafferentation and was totally abolished when these treatments were combined. These results suggest that gastric HCO(3)(-) secretion in response to acid is regulated by two independent mechanisms, one mediated by prostaglandins (PGs) and the other by sensory neurons and nitric oxide (NO). The acid-induced HCO(3)(-) secretion in the normal stomach is totally mediated by endogenous PGs, but, when the stomach is made slightly permeable to acid, the response is markedly facilitated by sensory neurons and NO. PMID- 16710056 TI - On the mechanism of some first-order enantiotropic solid-state phase transitions: from Simon through Ubbelohde to Mnyukh. AB - The first (so-called) lambda transition in solids was found in the specific heat measurements for NH(4)Cl at 242 K by F. Simon in 1922 [Simon (1922). Ann. Phys. 68, 241-280]. Analogous phenomena found in many other solids gave rise to doubts (expressed most clearly by A. R. Ubbelohde some 50 years ago) about the applicability of classical thermodynamics to some phase transitions [Ubbelohde (1956). Brit. J. Appl. Phys. 7, 313-321]. However, Y. Mnyukh's studies of enantiotropic phase transitions in eight organic crystals showed that all proceed by a nucleation-and-growth mechanism [summarized in Mnyukh (2001), Fundamentals of Solid State Phase Transitions, Ferromagnetism and Ferroelectricity. 1st Books]. Nucleation is localized at defects in the parent phase; growth can be epitaxic and oriented if parent and daughter phases have closely similar structures, or random (not oriented) if there are substantial structural differences. This conclusion is supported by a critical review of Mnyukh's eight examples and other results published in the interim. It seems that Ubbelohde's invocation of "hybrid crystals" and "smeared transitions" can mostly be accounted for by lack of equilibrium in the phase-transition studies cited by him. However, the intermediate phase in 4,4'-dichlorobenzophenone appears to have structural resemblances to Ubbelohde's' "hybrid crystal". PMID- 16710055 TI - Calcium waves in intact guinea pig gallbladder smooth muscle cells. AB - Intracellular Ca(2+) waves and spontaneous transient depolarizations were investigated in gallbladder smooth muscle (GBSM) whole mount preparations with intact mucosal layer [full thickness (FT)] by laser confocal imaging of intracellular Ca(2+) and voltage recordings with microelectrodes, respectively. Spontaneous Ca(2+) waves arose most often near the center, but sometimes from the extremities, of GBSM cells. They propagated regeneratively by Ca(2+)-induced Ca(2+) release involving inositol 1,4,5-trisphosphate [Ins(1,4,5)P(3)] receptors and were not affected by TTX and atropine (ATS). Spontaneous Ca(2+) waves and spontaneous transient depolarizations were more prevalent in FT than in isolated muscularis layer preparations and occurred with similar pattern in GBSM bundles. Ca(2+) waves were abolished by the Ins(1,4,5)P(3) receptor inhibitors 2 aminoethoxydiphenyl borate and xestospongin C and by caffeine and cyclopiazonic acid. These events were reduced by voltage-dependent calcium channels (VDCCs) inhibitors diltiazem and nifedipine, by PLC inhibitor U-73122, and by thapsigargin and ryanodine. ACh, CCK, and carbachol augmented Ca(2+) waves and induced Ca(2+) flashes. The actions of these agonists were inhibited by U-73122. These results indicate that in GBSM, discharge and propagation of Ca(2+) waves depend on sarco(endo)plasmic reticulum (SR) Ca(2+) release via Ins(1,4,5)P(3) receptors, PLC activity, Ca(2+) influx via VDCCs, and SR Ca(2+) concentration. Neurohormonal enhancement of GBSM excitability involves PLC-dependent augmentation and synchronization of SR Ca(2+) release via Ins(1,4,5)P(3) receptors. Ca(2+) waves likely reflect the activity of a fundamental unit of spontaneous activity and play an important role in the excitability of GBSM. PMID- 16710057 TI - Structure determination of A2M3+TaO6 and A2M3+NbO6 ordered perovskites: octahedral tilting and pseudosymmetry. AB - The room-temperature crystal structures of six A(2)M(3+)M(5+)O(6) ordered perovskites have been determined from neutron and X-ray powder diffraction data. Ba(2)YNbO(6) adopts the aristotype high-symmetry cubic structure (space group Fm?overline 3m, Z = 4). The symmetries of the remaining five compounds were lowered by octahedral tilting distortions. Out-of-phase rotations of the octahedra about the c axis were observed in Sr(2)CrTaO(6) and Sr(2)GaTaO(6), which lowers the symmetry to tetragonal (space group = I4/m, Z = 2, Glazer tilt system = a(0)a(0)c(-)). Octahedral tilting analogous to that seen in GdFeO(3) occurs in Sr(2)ScNbO(6), Ca(2)AlNbO(6) and Ca(2)CrTaO(6), which lowers the symmetry to monoclinic (space group P2(1)/n, Z = 2, Glazer tilt system = a(-)a( )c(+)). The Sr(2)MTaO(6) (M = Cr, Ga, Sc) compounds have unit-cell dimensions that are highly pseudo-cubic. Ca(2)AlNbO(6) and Ca(2)CrTaO(6) have unit-cell dimensions that are strongly pseudo-orthorhombic. This high degree of pseudosymmetry complicates the space-group assignment and structure determination. The space-group symmetries, unit-cell dimensions and cation ordering characteristics of an additional 13 compositions, as determined from X ray powder diffraction data, are also reported. An analysis of the crystal structures of 32 A(2)MTaO(6) and A(2)MNbO(6) perovskites shows that in general the octahedral tilt system strongly correlates with the tolerance factor. PMID- 16710058 TI - Structure prediction of ordered and disordered multiple octahedral cation perovskites using SPuDS. AB - The software package SPuDS has previously been shown to accurately predict crystal structures of AMX(3) and A(1 - x)A'(x)MX(3) perovskites that have undergone octahedral tilting distortions. This paper describes the extension of this technique and its accuracy for A(2)MM'X(6) ordered double perovskites with the aristotype Fm?overline 3m cubic structure, as well as those that have undergone octahedral tilting distortions. A survey of the literature shows that roughly 70% of all ordered double perovskites undergo octahedral tilting distortions. Of the 11 distinct types of octahedral tilting that can occur in ordered perovskites, five tilt systems account for approximately 97% of the reported structures. SPuDS can calculate structures for the five dominant tilt systems, Fm?overline 3m (a(0)a(0)a(0)), I4/m (a(0)a(0)c(-)), R?overline 3 (a(-)a( )a(-)), I2/m (a(0)b(-)b(-)) and P2(1)/n (a(-)a(-)b(+)), as well as two additional tilt systems, Pn?overline 3 (a(+)a(+)a(+)) and P4/mnc (a(0)a(0)c(+)). Comparison with reported crystal structures shows that SPuDS is quite accurate at predicting distortions driven by octahedral tilting. The favored modes of octahedral tilting in ordered double perovskites are compared and contrasted with those in AMX(3) perovskites. Unit-cell pseudosymmetry in Sr- and Ca-containing double perovskites is also examined. Experimentally, Sr(2)MM'O(6) compounds show a much stronger tendency toward pseudosymmetry than do Ca(2)MM'O(6) compounds with similar tolerance factors. PMID- 16710059 TI - "Ionic" size differences from bond-valence parameters and from ionic radii. AB - Expectation values for size differences between pairs of "ions" (atoms in oxidation states =/= 0) embedded into similar coordination environments are calculated from tables of bond-valence parameters and ionic radii. PMID- 16710060 TI - Experimental versus expected halide-ion size differences; structural changes in three series of isotypic bismuth chalcogenide halides. AB - Experimentally determined halide-ion size differences are compared with expected size differences in the three series of isotypic bismuth chalcogenide halide compounds, KBi(6)O(9)X (X = Cl, Br and I), BiOX (X = F, Cl, Br and I) and BiSX (X = Cl, Br and I). The strong deviations observed can be assigned to steric strain caused by the heterogeneity of the bond-valence pattern and (for BiOX) to anion anion repulsion and a change in the connectivity scheme. Some special features of the BiOF structure and the question of "isotypism" within the BiOX series are briefly discussed. Structural changes within the BiSX series are analysed. PMID- 16710062 TI - Estimation of polyhedral compressibilities and structural evolution of GdFeO3 type perovskites at high pressures. AB - A new approach based on the bond-valence matching relation is developed to predict the detailed structural evolution of GdFeO(3)-type perovskites at high pressure from knowledge of the room-pressure structure and the high-pressure unit cell parameters alone. The evolution of perovskite structures estimated in this way is in good agreement with the structure refinements available from high pressure single-crystal diffraction measurements of a number of perovskites. The method is then extended to predict the structure of MgSiO(3) perovskite at pressures for which no single-crystal structural data are available and the results are compared to ab initio quantum calculations of MgSiO(3) perovskite up to 120 GPa. PMID- 16710061 TI - Compressibility of the nitridosilicate SrYb[Si4N7] and the oxonitridoaluminosilicates MYb[Si4-xAlxOxN7-x] (x = 2; M = Sr, Ba). AB - The compressibilities of the nitridosilicate SrYb[Si(4)N(7)] and the oxonitridoaluminosilicates MYb[Si(4-x)Al(x)O(x)N(7-x)] (x = 2; M = Sr, Ba) were investigated by in situ high-pressure X-ray powder diffraction. Pressures up to 42 GPa were generated using the diamond-anvil cell technique. The title compounds are structurally stable to the highest pressure obtained. A fit of a third-order Birch-Murnaghan equation-of-state to the p-V data results in V(0) = 302.91 (6) A(3), B(0) = 176 (2) GPa and B' = 4.4 (2) for SrYb[Si(4)N(7)]; V(0) = 310.4 (1) A(3), B(0) = 161 (2) GPa and B' = 4.6 (2) for SrYb[Si(4-x)Al(x)O(x)N(7-x)]; and V(0) = 317.3 (5) A(3), B(0) = 168 (2) GPa and B' = 4.7 (2) for BaYb[Si(4 x)Al(x)O(x)N(7-x)]. While the linear compressibilities of the a and c axes of BaYb[Si(4-x)Al(x)O(x)N(7-x)] are very similar up to 30 GPa, distinct differences were observed for SrYb[Si(4)N(7)] and SrYb[Si(4-x)Al(x)O(x)N(7-x)], with the c axis being the most compressible axis. In all of the investigated compounds the bulk compressibility is dominated by the compression behaviour of the tetrahedral network, while the size of the substituted cation plays a minor role. PMID- 16710063 TI - Na2Si3O7: an incommensurate structure with crenel-type modulation functions, refined from a twinned crystal. AB - The structure of metastable, incommensurately modulated Na(2)Si(3)O(7) has been determined from single-crystal X-ray diffraction data. In contrast to previous investigations which stated that the compound crystallizes in an orthorhombic space group, this study shows that the compound is monoclinic with a pseudo orthorhombic cell and is affected by twinning. The structure is described in the (3 + 1)-dimensional superspace. Crenel-type modulation functions are used to account for an aperiodic sequence of right- and left-handed zweier single chains of silicate tetrahedra. The modulation mainly affects one of the two symmetrically independent tetrahedral chains, which are connected to build up [Si(3)O(7)](2-) layers. Sodium cations are coordinated by five oxygen ligands and provide linkage between adjacent tetrahedral sheets. Distortions of the silicate tetrahedra and crystal chemical relationships of the title compound to sodium and lithium di- and metasilicates are discussed in detail. PMID- 16710064 TI - Structural complexity in minerals: twinning, polytypism and disorder in the crystal structure of polybasite, (Ag,Cu)16(Sb,As)2S11. AB - The crystal structures of 222- and 221-polybasite [(Ag,Cu)(16)(Sb,As)(2)S(11)] crystals have been solved and refined by means of X-ray diffraction data (collected at 100 and 120 K, respectively) from twinned crystals. Both structures consist of the stacking of [(Ag,Cu)(6)Sb(2)S(7)](2-) and [Ag(9)CuS(4)](2+) module layers in which Sb forms isolated SbS(3) pyramids typically occurring in sulfosalts; copper links two S atoms in a linear coordination and silver occupies sites with coordination ranging from quasi-linear to almost tetrahedral. An Ag - > Cu substitution in the [(Ag,Cu)(6)Sb(2)S(7)](2-) module layer is observed in both structures, the substitution amount being larger in the 221- than in the 222 polybasite. A pattern of the possible mechanism regulating the type of unit cell that is stabilized is proposed: starting from the hypothetical stoichiometric and fully ordered Ag(15)CuSb(2)S(11) 222-polybasite structure, with a low C2/c monoclinic symmetry and a large 222 supercell, the disorder introduced by the substitution of Cu for Ag increases the symmetry with a cell reduction along the c axis yielding the 221 supercell and a trigonal crystal system. A further increase of the substitution gives rise to a folding of the cell along the a and b axes and the 111-pearceite structure, space group P(bar)3m1. PMID- 16710065 TI - A comparative crystallochemical analysis of binary compounds and simple anhydrous salts containing pyramidal anions LO3 (L = S, Se, Te, Cl, Br, I). AB - A comparative analysis of binary compounds and 61 simple anhydrous salts M(y)(LO(3))(z) (L = S, Se, Te, Cl, Br, I) was performed using the crystallochemical program package TOPOS. A topological similarity was found between the salts and six types of binary compounds (NaCl, NiAs, PoCl(2), Tl(2)S(2), ZnTe, rutile). It is notable that these structure relationships are typical for other groups of inorganic salts: borates, carbonates, nitrates, orthophosphates, orthoarsenates, sulfates, selenates, perchlorates, molybdates and halogenides of d-metals. For all the M(y)(LO(3))(z) compounds the topology and uniformity of the ion arrays were investigated. It has been established that in 36 out of the 61 salts at least one ion array has the topology of close packing or the body-centred cubic lattice. The results obtained have allowed us to come to conclusions about the structure-forming role of the arrays of various chemical composition. PMID- 16710066 TI - Crystal structures of the trifluoromethyl sulfonates M(SO3CF3)2 (M = Mg, Ca, Ba, Zn, Cu) from synchrotron X-ray powder diffraction data. AB - The crystal structures of divalent metal salts of trifluoromethyl sulfonic acid ("trifluoromethyl sulfonates") M(SO(3)CF(3))(2) (M = Mg, Ca, Ba, Zn, Cu) were determined from high-resolution X-ray powder diffraction data. Magnesium, calcium and zinc trifluoromethyl sulfonate crystallize in the rhombohedral space group R(bar)3. Barium trifluoromethyl sulfonate crystallizes in the monoclinic space group I2/a(C2/c) and copper trifluoromethyl sulfonate crystallizes in the triclinic group P(bar)1. Within the crystal structures the trifluoromethyl sulfonate anions are arranged in double layers with the apolar CF(3) groups pointing towards each other. The cations are located next to the SO(3) groups. The symmetry relations between the different crystal structures have been analysed. PMID- 16710067 TI - Cis/trans isomers of PtX2L2 (X = halogen, L = neutral ligand); the crystal structure of trans-dichlorobis(dimethyl sulfide)platinum(II) and the pressure dependence of its unit-cell dimensions. AB - trans-PtCl(2)(dms)(2) (dms is dimethyl sulfide) crystallizes in the space group P2(1)/n and adopts the molecular point group C(i), which is the most frequently occurring point group for trans-PtX(2)L(2) complexes (78%), as observed in the Cambridge Structural Database (CSD; 2005 release), followed by C(1) (16%). Density functional theory calculations show that the observed geometry for trans PtCl(2)(dms)(2) has slightly higher energy than the most favorable geometry in the point group C(2h), but this geometry would require a space group that hampers close packing, thus showing that intermolecular forces determine the point group for the title compound. High-pressure powder diffraction studies of trans PtCl(2)(dms)(2) show no phase transformation up to 8.0 GPa. The bulk modulus is 8.1 (6) GPa and the pressure derivative 8.1 (4). In the CSD, the number of cis- and trans-PtX(2)L(2) compounds are almost equal, viz. 156 cis and 160 trans compounds, showing no preference for either isomer in the solid state. PMID- 16710068 TI - X-ray crystallographic structures of enamine and amine Schiff bases of pyridoxal and its 1:1 hydrogen-bonded complexes with benzoic acid derivatives: evidence for coupled inter- and intramolecular proton transfer. AB - Crystal structures of Schiff bases containing pyridoxal (PL), N-(pyridoxylidene) tolylamine, C(15)H(16)N(2)O(2) (I), N-(pyridoxylidene)-methylamine, C(9)H(12)N(2)O(2) (III), and their 1:1 adduct with 2-nitrobenzoic acid, (I)(+) C(7)H(4)NO_4;- (II), and 4-nitrobenzoic acid, (III)(+) C(7)H(4)NO_4;- (IV), serve as models for the coenzyme pyridoxal-5'-phosphate (PLP) in its PLP-dependent enzymes. These models allow the study of the intramolecular OHN hydrogen bond of PL/PLP Schiff bases and the H-acceptor properties of their pyridine rings. The free base (I) forms hydrogen-bonded chains involving the hydroxyl side groups and the rings of adjacent molecules, whereas (III) forms related hydrogen-bonded cyclic dimers. The adducts (II)/(IV) consist of 1:1 hydrogen-bonded complexes, exhibiting strong intermolecular bonds between the carboxylic groups of the acids and the pyridine rings of (I)/(III). In conclusion, the proton in the intramolecular O-H...N hydrogen bond of (I)/(III) is located close to oxygen (enolamine form). The added acids protonate the pyridine ring in (II)/(IV), but only in the latter case does this protonation lead to a shift of the intramolecular proton towards the nitrogen (ketoimine form). All crystallographic structures were observed in the open form. In contrast, the formation of the pyridinium salt by dissolving (IV) leads to the cyclic aminal form. PMID- 16710069 TI - S,S-1,2-Dicyclohexylethane-1,2-diol and its racemic compound: a striking exception to Wallach's rule. AB - The structures of enantiopure S,S-1,2-dicyclohexylethane-1,2-diol and its racemic compound (rac-S,S-1,2-dicyclohexylethane-1,2-diol) have been determined at 295 and 173 K. The crystals of the enantiopure material are more than 4% denser than the crystals of the racemic compound, but the melting points indicate that the crystals of the less dense racemic compound are considerably more stable than those of the racemic conglomerate. This apparent exception to the correlation of crystal density and melting point is explained. The enantiopure crystals have four molecules in the asymmetric unit (Z' = 4). Two of the molecules have the conformation observed for the one independent molecule of the racemic compound and two have a higher energy conformation; the overall P2(1) structure is a perturbed version of a P2(1)2(1)2(1) structure with Z' = 2. The enantiopure and racemic crystals have the same hydrogen-bonding motif, but the motif in the former appears to be significantly strained. A reason why crystals of enantiopure material might be systematically less dense than crystals of its racemic compound and to be more likely to have Z' > 1 is suggested. PMID- 16710070 TI - 17 salts of ephedrine: crystal structures and packing analysis. AB - The structures of two neutral and 17 salt forms of the base (1R, 2S)-(-) ephedrine are reported. These structures are discussed in the light of the conformers of the ephedrine moiety, the existence of bilayers and the structure determining role of the counterions. Overall, most of the salt structures are essentially derived from the observed packing of the neutral base and are dominated by the amphiphilic nature of the ephedrine molecular structure. In a few cases the size and hydrophobicity of the counterion disrupts this behaviour. PMID- 16710071 TI - The incommensurately modulated structure of a tricyclic natural-product-like compound of empirical formula C22H20O3. AB - 3,4-Diphenyl-2a,5a,6,7,8,8a,8b-heptahydro-furo[4,3,2-de]chromen-2-one (1) was prepared as part of a project aimed at the synthesis of polycyclic natural product-like scaffolds. X-ray analysis of crystals grown from ethanol revealed an incommensurately modulated structure. Data include main and satellite reflections up to second order. The modulation vector was refined using the NADA program. The modulated character of the structure of the organic compound C(22)H(20)O(3) is interpreted in terms of the intermolecular C-H...O hydrogen bonds and close contact approximation. PMID- 16710072 TI - Chemical bonding in pentaerythritol at very low temperature or at high pressure: an experimental and theoretical study. AB - Chemical bonding in the pentaerythritol crystal based on the experimental electron density at 15 (1) K, and theoretical calculations at the experimental molecular geometries obtained at room and low (15 K) temperatures have been analyzed and compared in terms of the topological analysis. Topological electron density features corresponding to the high-pressure (1.15 GPa) geometry are also reported. In addition to the bond critical points (CPs) within the molecular layers, CPs between the atoms of different molecular layers have been located and the bonding character of these relatively weak interactions discussed. Atomic charges and energies have been integrated over the atomic basins delimited by the zero-flux surfaces, and the intermolecular interaction energies have been calculated. The interaction between molecular layers in the crystal becomes stronger both at very low temperature and high pressure, as demonstrated by the more negative intermolecular interaction energies, higher electron density and energy density values at the CPs, and sharper electronic-energy density profiles. PMID- 16710073 TI - Safety and efficacy of low-dose pioglitazone (7.5 mg/day) vs. standard-dose pioglitazone (15 mg/day) in Japanese women with type 2 diabetes mellitus. AB - It is well known that pioglitazone, a potent thiazolidinedione, improves metabolic control. However, weight gain or peripheral edema may be of major clinical concern when using this agent. The purpose of our study was to prospectively evaluate the effects of low-dose pioglitazone (7.5 mg/day) on metabolic control, weight gain and the incidence of edema compared with a standard dose of pioglitazone (15.0 mg/day) in patients with type 2 diabetes mellitus (T2DM). Ninety-five Japanese female patients (mean age 58.4 +/- 10.4 years) with newly diagnosed T2DM were selected for this study. They were randomly divided into the following 2 groups according to therapy regimens, and examined every month for 6 months after diagnosis. Group A consisted of 54 patients treated with low-dose pioglitazone orally; Group B, the control-group, consisted of 41 patients treated with standard-dose pioglitazone orally. The incidence of peripheral edema was significantly much lower in group A (2/54) than in group B (11/41) (p = 0.0014). In addition, % change of body weight during the 6-month treatment in group A was significantly less than that in group B (p < 0.0001). On the other hand, the % change of biochemical parameters including HbA1c did not differ significantly between group A and group B, although glucose and lipid control significantly improved from baseline in both groups. Our results demonstrate the safety and efficacy of low-dose pioglitazone, suggesting that it could be another good choice of treatment for Japanese women with T2DM. PMID- 16710074 TI - Expression of calcitonin receptor in rat mammary gland during lactation. AB - Calcitonin (CT) and calcitonin receptor (CTR) have been reported to play an important role in mammary tissue during pregnancy, lactation, and involution. In the present study, the expression and distribution of CTR mRNA in rat mammary tissue during pregnancy and lactation were investigated. As measured by real-time RT-PCR, CTR mRNA levels were increased only slightly during pregnancy, but increased markedly immediately postpartum and remained elevated through lactation, with the highest levels observed 14 days postpartum. In situ hybridization analysis showed that intense CTR mRNA signals were detected in the whole mammary gland. We performed immunohistochemistry to determine distribution of CTR in the mammary epithelium. CTR has been reported to act as an amylin receptor when heterodimerized with receptor activity modifying protein-1 (RAMP1) or RAMP3. mRNA expression of RAMP1 and RAMP3 in mammary tissue decreased during pregnancy and lactation, and amylin mRNA was undetectable, suggesting that up regulated CTR in lactating mammary tissues binds CT rather than amylin. In primary cultures of mammary cells isolated from rat dams 14 days postpartum, CT produced a statistically significant decrease in thymidine incorporation. These results suggest that up-regulation of CTR during lactation may contribute to inhibition of mammary epithelial cell proliferation. PMID- 16710075 TI - An instructive case suggesting cyclical primary hyperparathyroidism. AB - We report an instructive case of primary hyperparathyroidism in which cyclical secretion of PTH may have caused repeated hypercalcemic crises followed by temporary remission with a spontaneous drop in PTH. A 64-year-old man was admitted to our hospital twice with severe hypercalcemic crisis (corrected calcium (cCa) 15.0 mg/dl and 16.7 mg/dl) accompanied by an increase in intact PTH (220 pg/ml and 470 pg/ml). During both events, the serum PTH values spontaneously dropped followed by remission of the hypercalcemia. The tumor, detected at the left-upper side, showed neither vascularity on ultrasound examination nor washout delay on MIBI scintigraphy, suggesting that two parathyroid adenoma infarctions had occurred. Cervical exploration was undertaken. The histopathological examinations confirmed that the tumor was parathyroid adenoma. Contrary to our expectation, however, it did not reveal necrotic tissue that would indicate recent infarction. The findings in this case may be explained by cyclical secretion of PTH from a parathyroid adenoma. Although cyclical Cushing's syndrome is well known, to our knowledge this is the first documented case suggesting cyclical primary hyperparathyroidism. PMID- 16710076 TI - Hormone profiles after intramuscular injection of testosterone enanthate in patients with hypogonadism. AB - To examine hormone levels after androgen replacement therapy (ART) in Japanese male patients with hypogonadism, nine Japanese male patients with hypogonadism (serum total testosterone (tT) or free testosterone (fT) levels of < or = 2.7 ng/mL or < or = 10 pg/mL, respectively; average age, 59 years) were enrolled. They were treated with 125 mg of testosterone enanthate by single intramuscular injection. Blood samples were collected on the morning of the day of treatment, pre-ART, as well as on days 1 to 7 and day 14 after administration. Serum levels of tT, fT, estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and sex hormone-binding globulin (SHBG) were determined. On day 1 after administration, the mean serum levels of tT and fT were 7.62 ng/mL and 23.22 pg/mL, respectively. Serum levels of tT and fT on day 14 after administration were lower than their pre-ART values. One patient exhibited abnormally high serum tT and fT levels of 19.6 ng/mL and 44.4 pg/mL, respectively. Serum levels of LH and FSH began to decrease gradually on day 5 after administration. Serum levels of SHBG did not change throughout the observation period. Serum levels of E2 increased 1.7 times on day 1 after administration but returned to its pre-ART value by day 14 after administration. The dose of testosterone enanthate for male patients with hypogonadism requiring ART should be determined carefully because some patients exhibited high serum levels of androgen beyond the physiological range and gonadotropin was suppressed in all treated patients. PMID- 16710077 TI - Evidence-based healthcare and health informatics: derivations and extension of epidemiology. AB - Epidemiology provides extremely valid information and evidence regarding human health. Epidemiologic findings with regard to major illnesses must be amassed, enhanced, and expanded further into related areas as a foundation for evidence based medicine that is based on clinical practice, as well as for evidence-based healthcare that includes public health-related issues. Epidemiology should be recognized not only by epidemiologists but also by a variety of people, including specialists in other areas for healthcare and medicine, people in law and media, policy makers, and the general public. A system is needed that can create information for facilitating appropriate decision-making with issues related to clinical medicine and public health. The principles and methodology of epidemiology are used as a base for developing a field of health informatics. The objective of health informatics is to establish a system for facilitating the flow and circulation of health and medical information. Health informatics has potential applications for the creation, communication, and use of information, and the discipline is being expanded as a practical applied science in search of solutions. This report represents an effort to expand the scope of health informatics and extend the applications of epidemiology by working with individuals in other disciplines and the public. PMID- 16710078 TI - Difference of progression to AIDS according to CD4 cell count, plasma HIV RNA level and the use of antiretroviral therapy among HIV patients infected through blood products in japan. AB - BACKGROUND: It is important to examine progression to acquired immunodeficiency syndrome (AIDS) or death and its predictors among human immunodeficiency virus (HIV) infected persons before and after the introduction of the highly active antiretroviral therapy (HAART) available in Japan since 1997. METHODS: The data used were from a survey of persons with HIV infected through blood coagulation factor products in Japan. Progression to AIDS or death during two periods, between January 1994 and March 1997, and between April 1997 and March 2002, were observed. RESULTS: The AIDS-free proportion after 3 years was 74% among 417 participants for the earlier period and 94% among 605 participants in the later one. The hazard ratio of low CD4 cell count (less than 200 cells/microL) was 50.8 for the earlier period and 4.7 for the later one compared with that of 500 cells/microL or more. After adjustment by plasma HIV RNA levels and use of antiretroviral therapy, the hazard ratios of the low CD4 cell count for the later period were still significant. CONCLUSION: The AIDS-free proportion among people with HIV infected through blood products in Japan largely increased after the introduction of HAART. The CD4 cell count remains an important predictor of future progression, but its importance might be less because of HAART. PMID- 16710079 TI - Impact of the revision of a nutrient database on the validity of a self administered food frequency questionnaire (FFQ). AB - BACKGROUND: Revision of the national nutrient database in 2000 had a strong impact on the absolute level of estimated nutrient intake in dietary assessments. However, whether it influenced the ranking of individuals by estimated intake, a more important function in epidemiologic studies, has not been investigated. Here, we investigated the effect of this revision of the nutrient database on the validity of a food frequency questionnaire (FFQ) used to estimate nutrient intake in the Japan Public Health Center-based prospective Study (JPHC Study). METHODS: Subjects were a subsample of the JPHC Study who volunteered to participate in the validation study of the FFQ. Validity of the FFQ was evaluated by reference to the 28-day weighed dietary records as a gold standard. Nutrient intake according to the FFQ was recalculated using the revised database, and the results were compared to those using the previous database. Spearman's rank correlation coefficients (CCs) between intakes estimated by the FFQ and dietary records were computed using the revised database, and were compared to CCs computed using the previous database. RESULTS: For most of the nutrients, mean intake increased or decreased significantly using the revised database. However, no notable change was seen for the CC between estimated intake according to dietary records and FFQ when the revised database was used for calculation. Differences in the point estimates of the CCs ranged from -0.14 to 0.15. Likewise, CCs between biomarkers and estimated intake according to FFQ were similar for the two databases. CONCLUSION: Despite changes in intake levels for many nutrients, the validity of our FFQ using rank correlation by nutrient intake was not influenced by revision of the nutrient database in Japan. PMID- 16710080 TI - Eating fast leads to obesity: findings based on self-administered questionnaires among middle-aged Japanese men and women. AB - BACKGROUND: Few epidemiologic studies have examined the association between the rate of eating and obesity. In this study, we cross-sectionally examined the association of the self-reported rate of eating with current Body Mass Index (BMI), and BMI-change from 20 years of age to the current age. METHODS: Subjects were 3737 male (mean age +/- standard deviation and mean BMI +/- standard deviation: 48.2 +/- 7.1 years and 23.3 +/- 2.7 kg/m(2)) and 1005 female (46.3 +/- 7.0 years and 21.8 +/- 2.8 kg/m(2)) Japanese civil servants. We measured self reported categorical rate of eating, current BMI, BMI at age 20, and BMI-change from age 20. Energy intake was assessed over a 1-month period with a brief-type diet history questionnaire. RESULTS: The multiple regression analysis in which the current BMI was regressed by categorical rate of eating, energy intake, age, and lifestyle factors showed that current BMI steadily increased by -0.99, -0.67, 0.81, and 1.47 kg/m(2) along with the progress of categorical rate of eating from the 'medium' group to 'very slow', 'relatively slow', 'relatively fast', and 'very fast' groups, respectively, in men. In women, the corresponding values were -1.06, -0.35, 0.50, and 1.34 kg/m(2). When the BMI increment from age 20 to current age was regressed in the same manner, the increment was -0.63, -0.34, 0.57, and 1.05 kg/m(2) in men and -0.71, -0.32, 0.34, and 1.14 kg/m(2) in women, respectively. Additionally, both BMI at age 20 and current height were positively associated with rate of eating. CONCLUSIONS: Our results among middle-aged men and women suggest that eating fast would lead to obesity. PMID- 16710081 TI - Dietary risk factors for colon and rectal cancers: a comparative case-control study. AB - BACKGROUND: In Japan, the incidence rate of colon cancer has more rapidly increased than that of rectal cancer. The differential secular trends may be due to different dietary factors in the development of colon and rectal cancers. METHODS: To compare dietary risk factors between colon and rectal cancers, we undertook a case-control study at Aichi Cancer Center Hospital, Japan. Subjects were 507 patients with newly diagnosed colon (n = 265) and rectal (n = 242) cancers, and 2,535 cancer-free outpatients (controls). Intakes of nutrients and food groups were assessed with a food frequency questionnaire, and multivariate adjusted odds ratios (ORs) were estimated using unconditional logistic models. RESULTS: We found a decreasing risk of colon cancer with increasing intakes of calcium and insoluble dietary fiber; the multivariate ORs across quartiles of intake were 1.00, 0.90, 0.80, and 0.67 (trend p = 0.040), and 1.00, 0.69, 0.64, and 0.65 (trend p = 0.027), respectively. For rectal cancer, a higher consumption of carotene and meat was associated with a reduced risk; the corresponding ORs were 1.00, 1.10, 0.71, and 0.70 for carotene (trend p = 0.028), and 1.00, 0.99, 0.68, and 0.72 for meat (trend p = 0.036). Carbohydrate intake was positively correlated with the risk of rectal cancer (ORs over quartiles: 1.00, 1.14, 1.42, and 1.54; trend p = 0.048). This association was stronger in women, while fat consumption was inversely correlated with the risk of female colon and rectal cancers. CONCLUSIONS: Dietary risk factors appear to considerably differ between colon and rectal cancers. PMID- 16710082 TI - Aspirin or anticoagulants in stenosis of the middle cerebral artery: A randomized trial. AB - BACKGROUND: We report the results of an open, randomized, multicenter trial that compared the efficacy of aspirin to oral anticoagulants (OA) for the prevention of vascular events in patients with symptomatic stenosis of the middle cerebral artery (MCA). METHODS: Participants were randomly assigned to receive 300 mg/day of aspirin or a dose of OA (target INR 2-3). The MCA stenosis was demonstrated by conventional angiography or by at least two noninvasive examinations. Patients had either transient ischemic attack or cerebral infarct (CI) attributable to the MCA stenosis within 90 days before inclusion. The primary endpoint was: nonfatal CI, nonfatal acute myocardial infarct, vascular death and major hemorrhage. The patients were followed-up for a minimum of 1 year and a maximum of 3 years. RESULTS: The study included 28 patients (14 in each treatment group); the average age was 67 +/- 9.9 years. Men constituted 68% of the patients. After a mean follow-up of 23.1 +/- 10.9 months, there were no recurrences of CI in both groups. No endpoint was reported in the aspirin group, but 2 patients in the OA group (14.3%) exhibited vascular events: 1 acute myocardial infarct and 1 intracerebral hemorrhage). However, this difference was not statistically significant (p = 0.48). CONCLUSIONS: Our study suggests that aspirin is the treatment of choice for the prevention of vascular events in patients with symptomatic MCA stenosis. PMID- 16710083 TI - Accuracy of conventional plus transoral carotid ultrasonography in distinguishing pseudo-occlusion from total occlusion of the internal carotid artery. AB - BACKGROUND: To investigate the accuracy of conventional carotid ultrasonography (CCU) combined with transoral carotid ultrasonography (TOCU) for distinguishing pseudo-occlusion from total occlusion of the internal carotid artery (ICA). METHODS: This study included 95 patients who were suspected of having an occlusion of the ICA on magnetic resonance angiography (MRA) and underwent both CCU and conventional digital subtraction angiography (DSA) in order to confirm the diagnosis. TOCU was also performed to observe the cervical portion of the ICA distal to the stenosis. We compared the ultrasonographic findings with the DSA findings. RESULTS: Twelve of the 95 patients were defined as having an ICA pseudo occlusion on DSA. On B-mode images with CCU color Doppler, slight residual flow signals in the ICA lumen were shown in 20 patients. Among them, 2 patients had a pulsed Doppler waveform of the distal ICA occlusion pattern. Among the remaining 18 patients, 4 had a pulsed Doppler waveform of the to and fro flow pattern, and 14 had a weak antegrade flow pattern in the ICA lumen. The conventional ultrasonographic method showed 100% sensitivity with 93% specificity for diagnosing an ICA pseudo-occlusion. The addition of TOCU findings increased the specificity to 98%. In 2 patients, who were overdiagnosed as having an ICA pseudo occlusion even using TOCU, DSA revealed an occlusion of the ICA distal to the ophthalmic artery with a severe stenosis of the proximal ICA. CONCLUSIONS: Using conventional and transoral carotid ultrasonography, an ICA pseudo-occlusion can be diagnosed with higher accuracy. PMID- 16710084 TI - Changes in brain volume 2 years after extracranial-intracranial bypass surgery: A preliminary subanalysis of the Japanese EC-IC trial. AB - BACKGROUND AND METHODS: We studied changes in brain volume on magnetic resonance imaging in 10 patients with and without extracranial-intracranial (EC-IC) bypass surgery who met the inclusion criteria for the Japanese EC-IC Bypass Trial, a multicenter, randomized, prospective study of patients with hemodynamic brain ischemia due to cerebral artery occlusive disease. We also examined the association of cerebral hemodynamics on single-photon emission computed tomography with the changes in brain volume. RESULTS: The affected/unaffected ratio of the percent brain volume declined in patients without EC-IC bypass surgery (p < 0.02, n = 4), and the affected/unaffected percent regional cerebral blood flow ratio increased in patients with the surgery (p < 0.03, n = 6). Acetazolamide reactivity increased in the affected hemisphere of patients with surgery (p < 0.01). Two-year increase (decrease) in acetazolamide reactivity of the affected hemisphere showed a significant positive correlation with 2-year changes in the affected/unaffected percent brain volume ratio (R2 = 0.737, p = 0.0007). CONCLUSIONS: Change in acetazolamide reactivity might be a good predictor for brain atrophy in cerebral artery occlusive disease. PMID- 16710085 TI - Early mobilisation after stroke: Review of the literature. AB - BACKGROUND: Mobilisation of acute stroke patients--in bed and out of bed as early as possible--is currently recommended to prevent general and neurological complications. However, mobilisation protocols are poorly defined and need to be standardised in order to evaluate their clinical benefits. The purpose of this study is to highlight recent advances in recommendations for mobilisation of acute stroke and early neurological rehabilitation as found in the recent literature. METHODS: A Medline research was conducted from January 1950 to August 2005. RESULTS: Data synthesis of the observational studies and meta-analyses suggests that for most patients with acute stroke, early mobilisation is not harmful and may well be beneficial. However, no controlled study exists comparing early (e.g. 1-3 days) versus late (e.g. 1-2 weeks) mobilisation. Based on the available data and on pathophysiological considerations, we have developed a partially individualised procedure with precise entry and exit points for early mobilisation according to clinical status. CONCLUSIONS: Insufficient data are available to prove the beneficial effects of early mobilisation after stroke, although early neurological rehabilitation as part of routine stroke unit care seems to contribute to good long-term outcome. This article presents a standardised procedure for early mobilisation that may be tested in a randomised study between early mobilisation out of bed (3 days) compared to late mobilisation (6 days). PMID- 16710086 TI - Effect of simvastatin on coronary lesion site remodeling: a serial intravascular ultrasound study. AB - BACKGROUND: Direct evidence of coronary artery remodeling can be derived only from serial changes in the external elastic membrane (EEM) and plaque area. The aim of the study was to assess the effect of simvastatin on coronary remodeling in serial intravascular ultrasound (IVUS) studies. METHODS: In 39 male patients ECG-triggered transducer pullback IVUS was performed at baseline, after 3 months on a lipid-lowering diet (control period), and after another 12 months of simvastatin 40 mg/day. The lesion site was the image slice with maximum plaque burden at 3 months. RESULTS: Absolute changes in the EEM area correlated significantly with changes in plaque area during the control period [B = 0.966, r = 0.792 (95% CI 0.71-1.22); p < 0.001] and during simvastatin treatment [B = 0.945, r = 0.822 (95% CI 0.73-1.16); p < 0.001], but there was no significant difference in the slope (delta EEM/delta plaque) between the two time intervals. After 12 months of simvastatin, there was a significant reduction in the lesion EEM area of 4.6% (p = 0.006) and in the lesion plaque area of 5.9% (p < 0.001), but there was no change in reference measurements. As a result, the remodeling index was reduced by simvastatin from 1.01 +/- 0.12 to 0.95 +/- 0.09 (p < 0.001). CONCLUSION: Simvastatin decreases the remodeling index by reducing lesion, but not reference plaque and EEM area. However, simvastatin does not affect direct evidence of remodeling (delta EEM/delta plaque) obtained using serial IVUS studies. PMID- 16710087 TI - Comparison of the test of everyday functional abilities with a direct measure of daily function. AB - OBJECTIVES: To compare scores for dementia patients on a newly developed direct test of instrumental activities of daily living (IADLs) to scores on a widely accepted functional measure. METHODS: In 27 community-dwelling dementia patients, scores for the Test of Everyday Functional Ability (TEFA) were compared with scores on a widely used direct test of daily function (Independent Living Scale; ILS). RESULTS: Mini-Mental State Examination scores ranged from 5 to 28 (mean +/- SD = 20.7 +/- 6.37). There was a highly significant correlation between total TEFA and total ILS scores (r(25) = 0.892, p < 0.001) as well as many of the comparable subscales. CONCLUSION: The TEFA is a valid measure of IADL competence for dementia patients. PMID- 16710088 TI - Dementia in subjects aged 75 years or over within the PAQUID cohort: prevalence and burden by severity. AB - BACKGROUND/AIMS: To analyze the prevalence of dementia by severity and to describe the sociodemographic characteristics of people with dementia in the community, as well as the consequences of this disease in terms of disability and institutionalization. METHODS: This study was based on the PAQUID community-based cohort study of 1,461 subjects aged 75 years or over. Severity of dementia was assessed using the Mini-Mental State Examination (MMSE). RESULTS: The prevalence of dementia was estimated to be 17.8%, with about 43% at a moderately severe or more severe stage of the disease (MMSE 40 years, pathological laughing crying, unemployment. CONCLUSIONS: Our brief battery is an easy and reproducible tool. Completed with warning signs indicating the need for neuropsychological screening, this tool provides the practitioner with a global means of assessing disease activity and potentially therapeutic efficacy. PMID- 16710127 TI - [Analysis of reasons for the late diagnosis of amyotrophic lateral sclerosis]. AB - INTRODUCTION: The mean diagnostic delay of amyotrophic lateral sclerosis (ALS) is greater than one year. Its causes are multiple, related to the affection, the patient, or medical practices. METHODS: An investigation was carried out in 77 consecutive patients, to describe their medical course since the date of the first symptoms until the diagnosis. Interrogation of the private practitioners potentially implied in the diagnostic procedure enabled an evaluation of the degree and origin of their knowledge of the disease. RESULTS: In the majority of patients, the first consultation was conducted by a general practitioner. Referral to a neurologist occurred on average 7 months after this consultation. A first-intention electrophysiological examination was prescribed in one-third of patients. Practitioners were unknowledgeable about certain clinical signs. Their knowledge on the disease came primarily from medical school training and for two thirds, their satisfaction concerning their degree of training was low. CONCLUSION: This observational study shows that medical practices contribute to delayed diagnosis of ALS, particularly delayed referral to a neurologist. To enable earlier diagnosis, general practitioners should be informed of the usefulness of early referral for multidisciplinary care of patients. PMID- 16710128 TI - [The presentation of viral hepatitis C infection without cryoglobulinemia with a peripheral neuropathy. Five case reports]. AB - INTRODUCTION: Peripheral neuropathies are the most common neurological complication of viral hepatitis C infection with mixed cryoglobulinemia. CASES REPORT: We report five cases (three men, two women) of peripheral neuropathies revealing viral hepatitis C infection without cryoglobulinemia; the patients' mean age was 56 years. Paresthesias were the most frequent symptom. Electroneuromyographic examination found one case of polyneuropathy and four cases of multiplex mononeuropathies; the complement level was normal in all patients and the rheumatoid factor positive in two cases. Etiological investigations for peripheral neuropathy remained negative. Treatment and outcome were variable. DISCUSSION: Negative cryoglobulinemia in cases of VHC infection with neurological features has been described in the last few years, suggesting the possibility of other mechanisms such as direct action of the virus on the nervous system. There is no consensus on the treatment and outcome is variable. CONCLUSION: Peripheral neuropathy may reveal VHC infection, underscoring the need for VHC serology testing in etiological investigations for peripheral neuropathies. PMID- 16710129 TI - [Identification errors and delusions of false identification in Alzheimer's disease: a regional survey]. AB - INTRODUCTION: Misidentification syndromes are the main symptoms in Alzheimer's disease. Underlain by complex cognitive, agnosic, and amnesic disturbances of degenerative etiology, they can be expressed by misidentification delusions, as in psychoses. To date, research has focused on identification disturbances of persons selected according to various definitions. OBJECTIVE: Our main objective was to evaluate the frequency of identification disturbances among patients suffering from Alzheimer's disease within the current conditions of diagnosis and treatment. The secondary objective aimed to establish a detailed analysis of symptoms and clinical correlations, and evaluate the effects of the troubles on the caregiver. METHODS: We conducted a regional survey using a questionnaire designed for the caregiver, proposed to the 60 geriatric doctors and neurologists in the Poitou-Charentes region of France, for all patients suffering from Alzheimer's disease (defined according to DSM IV criteria) seen between June 1st and August 31st of 2003. Statview software was used for statistical analysis. RESULTS: The survey was completed for 104 patients: 69.5 percent women and 30.5 percent men, with a mean age of 79 years. The majority of the patients were seen by a neurologist and presented an average cognitive deficiency (MMS ranging from 11 to 20). An identification disturbance, whether or not it was delusional, all domains included, was found in 81.6 percent of the patients and was related to a more severe cognitive deficiency and greater hardship on the part of the caregiver. DISCUSSION: We observed a high prevalence of identification disturbance, which can be explained by the systematic and exhaustive search for identification disturbance. The most frequent disturbance concerned the identification of places, whereas self-identification was less often affected; the authentic Capgras delusion was found less often. CONCLUSION: The study of identification disturbances in Alzheimer's disease can contribute to a better understanding of the cognitive, psychopathological, and physiopathological aspects of the disease as well as to a better knowledge and better care for the patient. PMID- 16710130 TI - [Acute bilateral deafness and facial diplegia as a presentation of occlusion of the basilar artery]. AB - INTRODUCTION: Sudden bilateral deafness and facial weakness are unusual presentations of brain stem stroke. OBSERVATION: We report the case of a patient who presented successively sudden bilateral deafness and facial diplegia in correlation with a brain stem stroke but without any ischemic pontine lesion. DISCUSSION: Unlike our case, all of the earlier publications, have reported the presence of ischemic pontine lesions in patients with bilateral deafness and facial diplegia. Selective vulnerability of inner ear to ischemia has been hypothesized but cannot explain the facial diplegia. CONCLUSION: Our case would suggest extra-neuraxis failure by ischemia of the acoustico-facial nerve. PMID- 16710131 TI - [Symptomatic periodic paralysis secondary to primary Sjogren's syndrome]. AB - INTRODUCTION: Hypokalaemic periodic paralysis can be primitive or secondary to potassium deficiency which can arise from several causes. Primary Sjogren's syndrome is a rare cause related to kidney involvement. CASE REPORT: A 50-year old woman has been admitted for hypotonic tetraparesis which had appeared a few days earlier. History taking revealed three previous similar episodes with a notion of oral and lacrimal dryness. Laboratory tests revealed severe hypokalaemia, hyperchloremia, alkaline urinary pH and a minima 24h proteinuria. Additional investigations led to the diagnosis of a primary Sjogren's syndrome defined on the basis of international criteria. Kidney biopsy revealed tubular interstitial nephritis. Oral corticosteroid therapy and potassium supplementation led to symptom improvement. A recurrent episode also responded to treatment. Additional urinary alkalinisation has prevented further relapse. DISCUSSION: Primary Sjogren's syndrome is an exocrine disease causing systemic disorders. Tubular-interstitial nephropathy may occur in 25 percent of patients leading to distal tubular acidosis defined by the association of hypokalaemia, hyperchloremia and alkaline urinary pH. When hypokalaemia is severe, periodic paralysis may occur. CONCLUSION: Primary Sjogren's syndrome can lead to nephropathy and subsequent hypokalaemic periodic paralysis. Urinary alkalinisation is essential to prevent this catastrophic presentation from recurring. PMID- 16710132 TI - [Pseudo-tumoral neuro-Behcet's disease]. AB - INTRODUCTION: Clinical involvement of the central nervous system occurs in about 10 to 30 percent of patients with Behcet's disease. Neurological pseudo tumoral presentation is rare. CASE REPORT: We report a case of pseudo tumoral neuro Behcet disease in a 38-year-old man, with past history of facial and dorsal folliculitis, who presented suddenly headache, right hemiplegia, aphasia and disturbed consciousness. Neuroradiological investigations showed a pseudo tumoral lesion in the left capsulo-thalamic region extending to the homolateral peduncle. The patient improved with steroid and immunosuppresseur therapy. CONCLUSION: The clinical and radiological presentation of neuro-Behcet's disease can mimic a brain tumor. PMID- 16710133 TI - [Coeliac disease an spastic paraplegia]. AB - INTRODUCTION: Celiac disease (CD) is an immune-mediated disease triggered by the ingestion of gluten in genetically susceptible individuals. Neurological manifestations are rare and severe and must be sought systematically. CLINICAL CASES: Two non related patients each from a consanguineous marriage developed progressive spastic paraplegia 2 and 8 years respectively after onset of CD. The radiological and biological findings were normal except for the presence of abnormalities related to CD. CONCLUSION: The relationship between spastic paraplegia and CD is not well established. Autoimmune, metabolic and genetic mechanisms could be considered but the probability of a fortuitous association should not be ruled out. PMID- 16710134 TI - [Epileptic seizures and vardenafil]. AB - INTRODUCTION: Epileptic seizures complicating treatment with selective inhibitors of phosphodiesterase type 5 are scarcely reported. CASE REPORT: A previously non epileptic 78-year-old patient presented with a partial epileptic seizure following oral intake, for the second time, of 10mg of vardenafil (Levitra). The brain MRI failed to show any preexisting lesion. To our knowledge, only 2 cases of generalized tonic-clonic seizures induced by sildenafil (Viagra) use have been reported. In our patient, the seizure could be due to the epileptogenic potential of the drug or to its vascular complications. CONCLUSION: Further studies are needed to elucidate the association of phosphodiesterase inhibitors use and epileptic seizures. PMID- 16710135 TI - [Report from the 58th annual meeting of the American Academy of Neurology]. PMID- 16710136 TI - [On the trivialization of cannabis and our responsibility as physicians]. PMID- 16710137 TI - [Suicide prevention: screening in the emergency department]. PMID- 16710138 TI - [Hospital-acquired pressure ulcers: avoidable disease or quality of care indicator?]. PMID- 16710139 TI - [Spontaneous pneumothorax: moving towards simplification]. PMID- 16710140 TI - [Peer visits to community-based general practitioners and pediatricians as part of the "Antibiotics only when necessary" campaign in the Alpes-Maritimes district. Methodology, feasibility, and interest]. AB - OBJECTIVES: As part of a public health campaign promoting prudent antibiotic use to combat bacterial resistance in southeastern France, a multidisciplinary group organized peer-conducted educational outreach (academic detailing) visits to all the community-based general practitioners and pediatricians in the district. The visits, which took place in 2000 and were repeated in 2003, were intended to provide epidemiological data and professional guidelines to these doctors. METHOD: A group of local experts created special academic detailing sheets based on French and international recommendations. Peers with similar experience received special training in the relevant scientific material and in communications skills and then visited the local physicians and discussed the detailing sheets. RESULTS: The project contacted 95% of the target doctors (1079/1174 in 2000 and 1119/1135 in 2003), 94% (1024/1079 and 1042/1119, respectively) of whom agreed to be visited. The visitors reported positive feedback in over 80% of cases. Many of the visited doctors showed keen interest that led to ongoing dialogue, including participation in a regional influenza surveillance program. A sample of the visited doctors received a questionnaire to evaluate the method in 2001: 93% of the responders endorsed the method and 92% requested for more recommendations and guidelines. DISCUSSION: Carefully prepared individual academic detailing visits conducted by peers succeeded in visiting nearly all the physicians in the district twice, two years apart, and provided them with useful support for implementing important guidelines in everyday practice. This method can be used to improve cooperation between healthcare providers and improve the quality of health care in France. PMID- 16710141 TI - [Chemical content of street cannabis]. AB - INTRODUCTION: There is little information available about the real content of cannabis consumed in France. OBJECTIVE: To assess the chemical content, including adulterants and contaminants, and potency of cannabis samples obtained from end users. METHODS: Samples of resins considered by users to produce "unusual" effects were collected from habitual consumers and chemically analyzed. RESULTS: Thirty-seven samples were obtained. Average THC content was 8.8%. The effects reported by users were consistent with the THC content. Apart from an unidentified fatty substance found in two samples, no adulteration was found. DISCUSSION: This study, which relied on volunteers, enabled us to collect samples considered abnormal by experienced users. It did not confirm the numerous anecdotes of adulteration. CONCLUSION: We suggest that this study should be widened to monitor the current chemical contents of cannabis derivatives available in France. The impact of providing users with the results remains to be evaluated. PMID- 16710142 TI - [Reduction in short-term repetition of attempted suicide associated with new organization of psychiatric management of suicide attempters. Two cohorts]. AB - OBJECTIVES: To investigate the impact of the reorganization of psychiatric care in the Accident and Emergency (A&E) unit on repeated suicide attempts. METHOD: The study includes suicide attempters admitted to the Rennes University Hospital (Brittany, France) A&E unit in 1994 (n=1003) and 2000 (n=1018). We compared the clinical data (age, gender, marital status, suicide history, methods, and outcome at discharge from A&E) and psychiatric management of the two cohorts. The rate of repeated suicide attempts was measured according to the same criterion in 1994 and 2000 (re-admission to the A&E unit). RESULTS: Both populations of suicide attempters were very similar from both a clinical and demographic point of view. The repetition rate at 1 month was significantly higher (OR=4.16, p=0.0001) in 1994 than in 2000. The difference between the cohorts continued at three and six months, but became statistically insignificant at one year. CONCLUSION: By 2000, organization of psychiatric care in the A&E unit had been substantially strengthened. The presence of a complete psychiatric team (nurses, junior psychiatrist, and senior psychiatrist) in the A&E unit, 24 hours a day, seven days a week, was associated with a lower rate of repeated suicide attempts. PMID- 16710143 TI - Needle aspiration as first-line management of primary spontaneous pneumothorax. AB - BACKGROUND: Initial management of primary spontaneous pneumothorax (PSP) remains a topic of debate. Recent guidelines recommend needle aspiration as the initial strategy for large PSP, but chest tube drainage is still widely used. METHODS: Over a six-year period, we used needle aspiration in all cases of large PSP at our center to assess this technique's efficacy in real-life clinical practice. Our prospective study in the pulmonary intensive care unit of a 680-bed university hospital included patients with a PSP greater than 20%, as estimated by the Light's index. Needle aspiration, performed after local anesthesia, used a plastic intravenous catheter. If it failed, we performed thoracic drainage. RESULTS: The study included 35 patients. The initial success rate of needle aspiration was 69% (n=24/35), the one-week success rate 63% (n=22/35), and the one-year rate 51% (n=18/35). Tolerance of needle aspiration was good except for transient vagal reaction in four patients. No risk factors (age, body mass index, delay before hospitalization, previous pneumothorax, or Light index at inclusion) predicted initial failure of needle aspiration. CONCLUSION: Our results confirmed that needle aspiration is an attractive therapeutic option for patients with large PSP: success was observed in approximately two thirds of cases. PMID- 16710144 TI - [Pressure sores in a university hospital]. AB - OBJECTIVES: To determine the prevalence of pressure sores, their risk factors, and the responsible microbial agents in an acute-care hospital and to evaluate their management. METHOD: A prevalence survey was conducted from 5 July through 9 July 2004. Investigators completed a standardized questionnaire for each hospitalized patient, including demographic data (age, sex, previous hospitalizations, etc.) and Braden scale risk factors (sensory perception, humidity, activity, mobility, nutrition, and friction and shear). Two experts in skin care detected pressure sores by physical examination of the patients. Each pressure sore was swabbed and inoculated on selective media. Management was evaluated by reviewing the clinical charts of each patient with a pressure sore. RESULTS: The study included 535 adult patients (aged 59 +/- 19 years): 75 ulcer sores were observed in 37 patients (prevalence=6.9%). Stage I sores accounted for 24% of the total, stage II for 29%, stage III 31%, and stage IV 16%. The most frequent site was the heel (41%), followed by the sacrum (20%), elbow (11%), back (7%) and ischial tuberosities (7%). Sixty (80%) were acquired while hospitalized. Age-adjusted multivariate analyses found that the risk factors significantly associated with pressure sores were Braden score< or =15 (OR=5.9, 95% CI: 2.4 13.7, p<0.0001) and previous pressure sores (OR=5.0 95% CI: 2.2-11.6, p<0.0001). Eleven sores (24.5%), mostly stage III and IV, were colonized by multiple-drug resistant bacteria (i.e., methicillin resistant Staphylococcus aureus, extended spectrum beta-lactamase Enterobacteriaceae). Seven (9.3%) of the 75 ulcers were diagnosed only during the survey, by the experts; of the 68 diagnosed before the survey, 57 (83.8%) had been under treatment. Treatment was considered inappropriate according to French guidelines in 31.6% of the cases. CONCLUSION: This prospective prevalence study resulted in better awareness of the patients at risk for pressure sores. It also made the recently created mobile geriatrics unit better known within the hospital. PMID- 16710145 TI - [Ocular manifestations of pseudoxanthoma elasticum]. AB - BACKGROUND: Pseudoxanthoma elasticum (PXE) is a hereditary disease characterized by skin, ocular, and cardiovascular lesions. Its prognosis depends mainly on the ocular and vascular lesions. METHODS: This study included 11 patients with PXE. All patients had an ophthalmologic examination. RESULTS: All patients had ocular lesions. Seven (63%) had angioid streaks and two, macular lesions. One patient had bilateral intrapapillary drusen (of the optic nerve head). CONCLUSION: Ocular lesions in PXE had substantial diagnostic value. Ocular complications affect functional prognosis. These patients must undergo routine ophthalmologic examinations to look for ocular manifestations and their complications. PMID- 16710146 TI - [Vertical maternal fetal transmission of the chikungunya virus. Ten cases among 84 pregnant women]. AB - INTRODUCTION: In March 2005, an epidemic of chikungunya virus began in the southern portion of Reunion Island (French overseas district in the Indian Ocean) and spread to the northern part of the island at the end of 2005. The Reunion South Hospital Group observed the first cases of pregnant women infected with the virus in June 2005. We report here for the first time maternal-fetal transmission of this virus. CASES: From June 2005 through the end of January 2006, 84 pregnant women had acute chikungunya infections during pregnancy. In 88% of these cases (n=74)--all involving infections relatively distant from delivery--the newborns appeared asymptomatic. Conversely, 10 newborns had severe attacks (4 with meningoencephalitis and 3 with intravascular coagulations) after birth and required prolonged neonatal hospitalization (6 in the neonatal intensive care unit with intubation and assisted ventilation). No infants died, but there was one case of severe intracerebral hemorrhage after severe thrombocytopenia. These cases were confirmed by specific serology testing or PCR or both for mothers and newborns. We note that all severe cases involved women with viremia and fever in the intrapartum period. PMID- 16710147 TI - [Spectacular suicide associated with mefloquine]. AB - INTRODUCTION: We present a case in which suicide was a severe neuropsychiatric reaction to treatment with mefloquine. Physicians must be aware of these serious psychiatric complications and bear them in mind when faced with atypical behavior or suspected suicide. CASE REPORT: The body of a 27-year-old man was discovered at his home, covered with multiple knife wounds. The autopsy report concluded that death was due to a craniocerebral wound from a violent blow. Homicide was initially suspected. Suicide during acute psychosis associated with mefloquine was suggested, and toxicologic analyses confirmed this hypothesis. DISCUSSION: Serious neurologic and psychiatric adverse events associated with mefloquine (Lariam) have been reported since its introduction in 1985. Mefloquine prophylaxis is recommended for travelers to high-risk areas of chloroquine resistant plasmodium falciparum. The risk of malarial infection and the proven efficacy of mefloquine to prevent malaria should be weighed against the risk of drug-associated adverse events. Physicians must nonetheless be aware of these serious psychiatric complications, especially when faced with atypical behavior and atypical suicides. The patient's' family and friends should be asked about a possible trips abroad that might have entailed antimalaria treatment, even several months earlier. Testing for mefloquine during toxicological examinations is then essential. The World Health Organization recommendations and contraindications must be followed in prescribing mefloquine. PMID- 16710148 TI - [Atypical course of falciparum malaria in an asplenic patient]. AB - INTRODUCTION: The aim of treatment for falciparum malaria therapy is to achieve adequate clinical and parasitologic response within 7 days of starting treatment, with no subsequent relapse. We report and discuss the atypical course of a falciparum malaria attack observed in an asplenic patient returning from Burkina Faso. CASE: This 34-year-old man was hospitalized for severe malaria and treated for 7 days with quinine, as an inpatient. Adequate early clinical response was observed. Twenty days after the end of the quinine course, he was readmitted for uncomplicated falciparum malaria. Inpatient treatment used mefloquine this time. Clinical response was adequate, but the blood smear was still positive 10 days later. It was decided not to administer further treatment at that time. Subsequent clinical and parasitologic assessments 1 month and 3 months later showed the patient was cured. CONCLUSION: This report illustrates the critical role played by the spleen in parasite clearance. Clinical and parasitologic assessments are essential after treatment of asplenic patients for falciparum malaria. PMID- 16710150 TI - [Behavioral and cognitive therapy for social phobia in a smoker]. AB - INTRODUCTION: Social phobias are associated with nicotine addiction, and treatment for them can enable the patient to stop smoking. CASE: A patient seeking treatment to stop smoking was found to have a social phobia, which was treated with behavioral and cognitive therapy. At the end of this treatment, the patient spontaneously and without aids stopped smoking. DISCUSSION: Social phobias may be a factor in the onset of smoking and in the difficulties and relapses associated with cessation. They must be treated. PMID- 16710149 TI - [Ophthalmic manifestations of lipoid proteinosis]. AB - INTRODUCTION: Lipoid proteinosis (LP), also known as hyalinosis (or lipoidosis) cutis et mucosae or Urbach-Wiethe disease, is a rare autosomal recessive disorder. It is associated with deposits of protein-lipid complexes in various tissues including the skin and mucous membranes. Ophthalmologic manifestations are frequent and can affect visual prognosis. CASE: This 28-year-old patient presented vesiculobullous lesions of the face that developed into varioloid scars associated with hoarseness. Ophthalmologic examination revealed unilateral lesions including hyaline deposits on the palpebral margins, iris, and trabecular zone, complicated by uveitis, cataract, and glaucoma, which caused the functional loss of the left eye. Histological examination of a cutaneous biopsy confirmed the diagnosis. DISCUSSION: Involvement of the eyelids is characteristic, and moniliform blepharosis is pathognomonic and frequent. This case featured a rare intraocular form (uveitis). Deposits may be found on the conjunctiva, cornea, trabecula and Bruch membrane. Conjunctival or cutaneous biopsy confirms the diagnosis. Available treatment is quite limited. PMID- 16710151 TI - [Efficacy of rituximab in hairy cell leukemia]. AB - INTRODUCTION: Rituximab is an alternative treatment for hairy cell leukemia, when the standard treatments are unavailable. CASE: An 82-year-old woman was diagnosed with hairy cell leukemia. The severity of her neutropenia ruled out purine analogs, while heart disease and age both contraindicated use of interferon. Rituximab, in four weekly treatments, was effective from the first treatment and the positive response was sustained thereafter. DISCUSSION: Rituximab may be used to treat hairy cell leukemia, especially for patients refractory to purine analogs or after relapse or in the case of severe neutropenia, when interferon is contraindicated. PMID- 16710153 TI - [Mental health in the workplace: a human and economic issue]. PMID- 16710152 TI - [Atypical crowned dens syndrome]. PMID- 16710154 TI - [Depression in the workplace]. AB - Depression is the object of a dense literature, and synthesizing it is more of a utopian ideal rather than a concrete possibility. Several specific risk factors for mental health are found in the workplace: work overloads, defective communications, role conflicts, competitive climate, and tolerance of violence. At the same time, few preventive measures have been implemented against mental disorders at work, nor are many protective factors present. One worker in ten suffers from depression, anxiety, stress, or overwork. To be distinguished from "burnout", depressive symptoms must induce clinically significant suffering with substantial deterioration in functioning at work. For depression to be recognized as a workplace accident, the employee must show that it was triggered by an unforeseen and sudden event (or at least one certainly) due to or at work. The causal link between an event at work and the depression must be shown (in particular by expert medical testimony about stress factors and indicators of vulnerability to depression). Its recognition as an occupational disease can be based on the presence of psychosocial factors described by models of workplace stress and on its description by the occupational physician. PMID- 16710155 TI - ["Drinking less is better". Combining early identification and brief intervention for patients at risk]. AB - Above 210 grams a week in men and 140 grams a week in women, alcohol consumption is a risk factor for avoidable mortality in the general population. Beyond specific risk situations (for example, pregnancy, medication that interferes with alcohol, operating machinery, or a history of alcohol-dependence) in which abstinence is recommended, consumption of levels below these thresholds (which represent respectively an average of 3 and 2 drinks a day) involves little risk. Above these thresholds, the frequency of secondary disease (principally cancers and cardiovascular, neurologic, hepatologic, and gastroenterologic disorders) contributes to reducing life expectancy in drinkers. Early identification of excessive but not dependent alcohol consumption is the only means of avoiding the morbidity and mortality associated with drinking. Health providers too often confound alcoholism with alcohol-related problems. Half of the deaths associated with alcohol, however, concern people who are not dependent on it. Excessive drinkers must be identified early if they are to be counseled and helped to reduce their consumption. The brief intervention is a counseling practice easy to learn. When practiced wisely (in people who drink to excess but are not alcohol dependent), this brief intervention takes 10 minutes and provides information, motivational and behavioral counseling. It can be learned in two evenings and is immediately transposable into daily practice. The brief intervention is effective. It leads to a reduction in consumption below the risk thresholds in 10 50% of cases. Any trained care giver in primary care, hospital, or preventive medicine can provide it. Tools for identification and intervention are available. Two screening questionnaires have been validated in French, the AUDIT (a self administered questionnaire) and the FACE (a questionnaire completed by the doctor). The procedures and philosophy of interventions are defined and validated, and training is available for all providers who want to acquire this practice. This original research activity leads to a new public health effort. WHO, the national association for prevention of alcoholism and addiction, and various public health agencies have developed the experimental program 'Drinking less is better', intended to adapt WHO tools for early identification and brief intervention (EIBI) to French medical practice. This research-activity was conducted in close association with its targets (especially general practitioners) and has contributed to defining the conditions for the diffusion of this EIBI in France. Based on its conclusions, the health authorities have launched a national training strategy. PMID- 16710156 TI - [Schizophrenia: on the genetic trail]. AB - Data from genetic epidemiology studies demonstrate the existence of hereditary factors in the genesis of schizophrenia. Most authors now think that a multigenic multifactorial model will explain the transmission of this disease. Several genes may be involved, either additively (their effects added together, each gene having its own contribution), or epistatically (expression of one gene effective only if another gene is also expressed). Other risk factors - environmental, biological, and neurodevelopmental - may also play a role. There is likely to be a threshold effect above which the sum total of these factors makes the disease clinically detectable. New study strategies have therefore sought to characterize candidate regions or genes based on various biochemical or neurodevelopmental hypotheses. This research collides, however, with the heterogeneity of symptoms as well as the diversity of the disease development. Other avenues of research therefore look towards the study of endophenotypes or 'markers of vulnerability' and towards the identification of genetically homogeneous subgroups of patients and families, among whom susceptibility genes may be detected. Current genetic studies agree about the complexity of the transmission of this disease with its heterogeneous polygenic characteristics. PMID- 16710157 TI - [Takayasu arteritis]. AB - Takayasu arteritis is an inflammatory arteritis affecting large vessels, predominantly the aorta and its main branches. Thickening of the vessel wall is an early hallmark of the disease and leads to stenosis, thrombosis, and sometimes aneurysm formation. Reported incidence ranges from 1.2 to 2.6/million/year. Women aged 20 to 40 are most likely to suffer from the disease than men. Manifestations are very polymorphous, with presentations ranging from asymptomatic to neurologic catastrophes. Prognosis depends essentially on complications (retinopathy, hypertension, aneurysm, aortic insufficiency) and initial disease aggressivity. Diagnosis is based on imaging methods. Doppler ultrasound, computed tomography, and magnetic resonance imaging are fast and reliable methods for assessing vessel anatomy and luminal status. Positron emission tomography with fluorodeoxyglucose appears to be a highly sensitive and effective method for detecting disease activity, especially since standard inflammatory markers seem ineffective. Until now, corticosteroids have been the treatment of choice. If remission does not occur, methotrexate is added. Percutaneous transluminal angioplasty and sometimes vascular surgery is necessary in cases of critical ischemia or threatening aneurysm. Duration of treatment, choice of second-line treatment, and protocol for tapering medication currently depend more on experience than on evidence based medicine. Multicenter studies are needed to guide future practice. PMID- 16710158 TI - [Primary immune deficiencies: also found in adults. A diagnosis to be aware of]. PMID- 16710159 TI - [Complement component deficiencies in human disease]. AB - The complement system, composed of several plasma and membrane proteins, is an integral part of the innate immune system and plays a role in inflammatory response, destruction of infectious agents, elimination of immune complexes, and control of the specific (adaptive) immune response. Hereditary deficiencies of complement components are relatively rare and associated with susceptibility to a wide variety of clinical diseases. Complement components may be target of antibodies (anti-C1q, factor H, C3 alternative convertase, or C3NeF autoantibodies or anti-C1 inhibitor antibodies) that lead to acquired deficiencies. Testing the complement system is especially necessary in patients with autoimmune diseases, some kidney diseases, recurrent infections (especially meningococcal), and angioedema. Precise clinical descriptions of the phenotypes associated with these deficiencies and their molecular diagnosis are necessary to improve our understanding of the role that the complement system plays in the physiopathological mechanisms of these diseases and to propose the most specific treatment for them. PMID- 16710160 TI - [Primary immune deficiencies in neutrophil functioning]. AB - Neutrophils are polynuclear cells essential to innate immunity. They are the first cells to migrate from the blood to the inflammatory site where they kill pathogens and secrete various mediators that regulate innate and adaptive immunity. Functional steps required for their microbicidal activity include: transendothelial migration, migration towards the invading pathogens, and then recognition, adhesion, engulfment, and killing of the target. Primary deficiencies of these stages are expressed by repeated and/or severe bacterial and fungal infections. These deficiencies include granule abnormalities and leukocyte adhesion deficiencies Type I and II, defective pathogen recognition and the defective oxidative burst that characterizes chronic granulomatous disease. PMID- 16710161 TI - [Mendelian susceptibility to mycobacterial disease: defects in the IL-12/IFNgamma pathway]. AB - Mendelian susceptibility to mycobacterial disease is a recently described entity, responsible for disseminated disease due to nonvirulent mycobacteria and, to a lesser extent, non-typhoid salmonella in otherwise healthy patients. Different mutations in 5 genes and allelic heterogeneity accounts for 12 different diseases. The proteins encoded by the mutated alleles all belong to the interferon gamma/interleukin 12 loop, a hallmark of granulomatous immune response. Patients with defects in the IFNgamma pathway are predisposed to mycobacterial diseases, while those with defects in the IL-12 pathway are threatened more often by non-typhoid (systemic) salmonellosis. Tuberculosis has been described in both of these signaling pathway defects. Genetic dissection of the IL-12/IFNgamma pathway should improve our understanding of the human immune response to mycobacteria and help us begin to elucidate the genetic bases of tuberculosis. PMID- 16710162 TI - [Adult-onset primary hypogammaglobulinemia]. AB - Hypogammaglobulinemia during adulthood can lead to recurrent infectious diseases, particularly pneumonia, otitis, and sinusitis, mainly due to Streptococcus pneumoniae and Haemophilus influenzae. Physicians must discriminate between primary and secondary hypogammaglobulinemia to provide appropriate treatment. Common variable immunodeficiency (CVID) is the most common cause of primary hypogammaglobulinemia diagnosed in adulthood. Clinical features include various combinations of infectious diseases, autoimmune diseases, lymphoproliferative disorders, and gastrointestinal diseases. Mutations in genes responsible for primary hypogammaglobulinemia in children are occasionally described in adults with CVID. ICOS and TACI deficiencies have recently been reported in a few CVID patients, although most cases have no proven genetic defects. The DefI cohort is recruiting adults with CVID throughout France. It will make it possible to describe the clinical, immunological, and genetic features of French patients with hypogammaglobulinemia and will help us to understand this immune deficiency better. PMID- 16710163 TI - [Diagnosis of lymphocytopenia]. AB - Lymphopenia is defined as a peripheral lymphocyte count lower than 1500/mm3 in adults and 4500/mm3 in children younger than eight months of age. We propose a classification of lymphopenia according to the mechanism involved: lymphocyte production defects, including primary immune deficiencies and immune deficiencies secondary to malnutrition or zinc deprivation; excess catabolism, due to causes including radiotherapy, chemotherapy, immunosuppressive therapy, HIV infection, and systemic lupus erythematosus; abnormal lymphocyte trapping, including mainly splenomegaly, certain viral infections, septic shock, extended burns, systemic granulomatosis, and corticosteroids; other causes of lymphocytopenia, with mechanisms that remain poorly understood: ethnicity (Ethiopians), lymphoma, renal insufficiency, and idiopathic CD4 lymphocytopenia. PMID- 16710164 TI - [Diagnosis of primary immunodeficiency in adult patients]. AB - The molecular bases of approximately one hundred primary immune deficiencies (PID) have been identified over the last 15 years. In adults, the diagnosis of PID cannot be evoked before ruling out acquired immunodeficiencies, which are far more frequent. The search for specific PIDs may be oriented by the type of agent responsible for severe and/or recurrent infection. More rarely, other clinical manifestations such as granulomatosis, autoimmune manifestations, hemophagocytic syndrome, lymphoproliferation, or solid tumors may also lead to the identification of PID. PMID- 16710165 TI - [Cardiac magnetic resonance imaging of tuberculous constrictive pericarditis]. PMID- 16710166 TI - Gene expression and discovery during lens regeneration in mouse: regulation of epithelial to mesenchymal transition and lens differentiation. AB - PURPOSE: It has been shown that after extracapsular lens removal by anterior capsulotomy in the mouse, the lens can be regenerated. However, as the capsular bag is filled with fibers, epithelial to mesenchymal transition (EMT), an event which is common after cataract surgery as well, takes place during early stages. This study, using a unique mouse model, was undertaken to identify novel regulators and networks in order to more clearly understand secondary cataracts at the molecular level. METHODS: We examined global gene expression via microarray analysis of mouse lens regeneration after extracapsular surgery. Gene expression at different times after surgery was correlated with the processes of EMT, which is seen in the initial stages of regeneration, and lens fiber differentiation, which occurs later. RESULTS: Several notable patterns were observed from the gene clustering data. It was obvious from the analysis that initially there is a response to injury, extensive matrix remodeling, and severe downregulation of genes encoding lens structural proteins. The patterns returned gradually to normal three weeks after surgery. New genes were identified from the clustering results that might be potential regulators of EMT and lens differentiation. CONCLUSIONS: With this approach, we demonstrated the utility of a mouse model to study secondary cataracts at the molecular level. Extension of these studies in mice with known mutations affecting EMT or lens differentiation should allow the identification of the crucial molecular players that could lead to better treatments of secondary cataracts. PMID- 16710167 TI - Screening of the MERTK gene for mutations in Japanese patients with autosomal recessive retinitis pigmentosa. AB - PURPOSE: To determine whether mutations in the MERTK gene are present in Japanese patients with autosomal recessive retinitis pigmentosa (arRP). METHODS: The coding sequence of all 19 exons and the adjacent flanking intron sequences of the MERTK gene were directly sequenced in 96 unrelated Japanese patients with arRP. RESULTS: Seventeen sequence variants were found; six missense changes, three isocoding changes, and eight intron changes were also observed. One arRP patient had a novel homozygous Leu12Pro missense mutation in the MERTK gene. CONCLUSIONS: Mutations in the MERTK gene are relatively rare in Japanese patients with arRP. PMID- 16710168 TI - Expression of Hsp70 and Hsp27 in lens epithelial cells in contused eye of rat modulated by thermotolerance or quercetin. AB - PURPOSE: To investigate the dynamic expression of heat shock protein (Hsp)70 and Hsp27 in lens epithelial cells (LECs) of contused eyes and the effects of heat shock and quercetin. METHODS: Rat eyes were randomized into three groups: contusion, heat shock, and quercetin. Eye contusions were induced by dropping a 20 g steel ball from a height of 20 cm for 100 times. Heat shock was induced by raising the core body temperature to 40.5-41.5 degrees C for 8 min, 2-3 h before eye contusion. Quercetin, an inhibitor of Hsp expression, was given via gavage (100 mg/kg) 2-3 h before eye contusion. Experimental eyes were enucleated at 0, 1, 3, 6, and 24 h following each group treatment for the detection of Hsp70 and Hsp27 expression in LECs using RT-PCR. Lens sections were examined by immunohistochemistry with antibodies to Hsp70. RESULTS: Basic expression of Hsp70 and Hsp27 was observed in LECs. Expression of Hsp70 in LECs was increased after contusion in a time-dependent manner. Preconditioning hyperthemia (45 degrees C, 8 min) resulted in a significant increase of Hsp70 expression compared to the base level, and enhanced the expression in each subgroup in contrast to the corresponding subgroup in the contusion group. However, Hsp70 levels in the 1-6 h subgroup in the quercetin group was significantly lower. There was no notable change of expression of Hsp27 between each group. Immunostaining of Hsp70 was prominent in LECs of contusion eyes and heat shock eyes compared with those in normal eyes. Immunostaining of Hsp70 was faint in LECs of quercetin eyes. CONCLUSIONS: Increased expression of Hsp70 in LECs of contused eyes may play a protective role against degeneration of lens protein. Thermal preconditioning possibly protects against lens injury by increasing the expression of Hsp70. Endogenous protective mechanisms may be important in eye contusion. PMID- 16710169 TI - The human cornea proteome: bioinformatic analyses indicate import of plasma proteins into the cornea. AB - Increased biochemical knowledge of normal and diseased corneas is essential for the understanding of corneal homeostasis and pathophysiology. In a recent study, we characterized the proteome of the normal human cornea and identified 141 distinct proteins. This dataset represents the most comprehensive protein study of the cornea to date and provides a useful reference for further studies of normal and diseased human corneas. The list of identified proteins is available at the Cornea Protein Database. In the present paper, we review the utilized procedures for extraction and fractionation of corneal proteins and discuss the potential roles of the identified proteins in relation to homeostasis, diseases, and wound-healing of the cornea. In addition, we compare the list of identified proteins with high quality gene expression libraries (cDNA libraries) and Serial Analysis of Gene Expression (SAGE) data. Of the 141 proteins, 86 (61%) were recognized in cDNA libraries from the corneas of dogs and rabbits, or humans with keratoconus, and 98 (69.5%) were recognized in SAGE data of mouse and human corneas. However, the percentages of identified genes in each of the protein functional groups differed markedly. Thus, exceptionally few of the traditional blood/plasma proteins and immune defense proteins that were identified in the human cornea were recognized in the gene expression libraries of the cornea. This observation strongly indicates that these abundant corneal proteins are not expressed in the cornea but originate from the surrounding pericorneal tissue. PMID- 16710170 TI - Systemic investigation of keratoepithelin deposits in TGFBI/BIGH3-related corneal dystrophy. AB - PURPOSE: To investigate the location and tissue-specificity of the pathologic keratoepithelin (KE) deposition in a patient with a keratoepithelinopathy (KEP), TGFBI/BIGH3-related corneal dystrophy. METHODS: An autopsy was performed in a patient with lattice type I corneal dystrophy (LCDI) after authorization was obtained from the family. Mutation screening in TGFBI/BIGH3 was done on the patient several years ago. Eighteen different tissues or organs, including brain, heart, lung, kidney, liver, lymph nodes, spleen, aorta, esophagus, bone marrow, urinary bladder (including a papillary urothelial carcinoma), samples of a metastatic squamous cell carcinoma, adrenal gland, parathyroid gland, muscle, prostate, and cornea were investigated, and sections from the tissues were labeled with KE2 rabbit TGFBI/BIGH3 antiserum. RESULTS: The patient, diagnosed with LCDI and Alzheimer's disease, died at 79 years of age from a complicated chronic obstructive lung disease. Mutation analysis showed the classical Arg124Cys mutation in exon 4 of TGFBI/BIGH3, associated with LCDI. Except for the cornea, immunostaining with KE2 antisera did not reveal any deposits in any of the 17 other organs analyzed. CONCLUSIONS: Pathologic deposits caused by KE accumulation were only observed in the cornea and in no other tissue or organ in this patient. These results suggest a cornea-specific mechanism in the aggregation of KE. Further studies need to be done to investigate whether the degradation of mutated KE generates cornea-specific fragments that aggregate or whether the clearing of normal fragments is different in affected corneas, which then leads to aggregation. PMID- 16710171 TI - Microglia and macrophages are increased in response to ischemia-induced retinopathy in the mouse retina. AB - PURPOSE: The ability of microglial cells (MG) and macrophages (MAC) to release cytokines, induce apoptosis, as well as perform phagocytic functions suggests a possible role in wound healing following oxygen-induced injury. This study was performed to determine the temporal and spatial expression of F4/80 (F4/80+) positive microglia/macrophages (MG/MAC) in areas of retinal damage in the mouse model of oxygen-induced retinopathy. METHODS: C57BL/6 postnatal day 7 (P7) mice were exposed to 75% O2 for 5 days (P12) then allowed to recover in room air. Hyperoxia-exposed (O2) mice (O2 refers to hyperoxia exposure from P7 to P12 only) were sacrificed on P12, P14, P17, and P21 and their eyes were examined. Localization of F4/80+ cells in FITC-dextran-perfused retinas allowed coordinate visualization of retinal vessels and MG/MAC via fluorescence microscopy. BrdU, a cellular proliferation marker, was injected intraperitoneally 1 h prior to sacrifice. Immunostaining was performed for a microglia and macrophage-specific antigen (F4/80) and BrdU. CCL2 (monocyte chemoattractant protein-1; MCP-1) expression was examined by quantitative real time reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: There was a marked increase (>500%) in MG/MAC in hyperoxia-exposed retinas on P17O2 and P21O2 compared to control retinas. At P17O2, MG/MAC were localized in areas of neovascularization (NV), revealing an intimate relationship between MG/MAC and neovascular tufts. However, P21O2 retinas demonstrated MG/MAC associated with avascular regions in the outer layers of the retina. Immunostaining for F4/80 and BrdU revealed rare co localization in hyperoxia-exposed retinas. Real time RT-PCR results demonstrated increased expression of CCL2 in P14O2- and P17O2- exposed retinas. CONCLUSIONS: Our results suggest that resident retinal microglia proliferation occurs at a low frequency in response to injury in this model. The substantial increase in total F4/80+ cells in hyperoxia-exposed retinas in conjunction with the upregulation of CCL2 is consistent with recruitment of hematogenous macrophages into the retina. The temporal and spatial localization of MG/MAC adjacent to neovascular tufts suggests these cells are modulating the retinal response to ischemia-induced retinopathy. PMID- 16710172 TI - Proteomic analysis of soluble factors secreted by limbal fibroblasts. AB - PURPOSE: To identify soluble factors selectively secreted by limbal fibroblasts as possible regulators of limbal basal epithelium. METHODS: Limbal, corneal, and conjunctival fibroblasts were first expanded in vitro in Dulbecco's modified Eagle medium containing 10% fetal bovine serum, and then maintained in serum-free medium for two weeks. Proteomic analysis of culture supernatants was done to compare differences in secreted matricellular proteins. Real time PCR and western blots were done to confirm the expression of secreted protein acid and rich in cysteine (SPARC), a protein found in abundance in extracellular proteins secreted by limbal fibroblasts. Immunohistochemistry of SPARC was done in human limbal tissue to show the spatial distribution of the protein. An adhesion assay was designed to demonstrate the effects of SPARC on an SV40 immortalized human corneal epithelial cell line (HCEC). RESULTS: Proteomic analysis revealed several proteins selectively secreted by limbal fibroblasts. The particular spots were identified as SPARC, vimentin, serine protease, collagen alpha 2 precursor, tissue inhibitor of metalloproteinase 2 (TIMP-2), and 5,10 methlenetetrahdrofolate reductase (FADH2). The expression of SPARC was confirmed by western blot analysis, and mRNA expression was significantly higher in limbal fibroblasts compared to central corneal fibroblasts when analyzed by real time PCR. Immunohistochemistry revealed higher distribution of SPARC in the subepithelial stroma of the limbus compared to the central cornea. The addition of 10 microg/ml murine SPARC in HCEC significantly reduced cell spreading at three h. CONCLUSIONS: The matricellular protein SPARC is preferentially secreted by limbal fibroblasts, and may modulate intercellular adhesion of basal limbal epithelial cells. PMID- 16710173 TI - [Morphological radioautographic study of the palatine tonsils in chronic tonsillitis and healthy subjects]. AB - A pilot morphological (radioautographic) study of healthy and affected palatine tonsils (in hypertrophy and toxic-allergic form of chronic tonsillitis) in children and adults has shown that intensity of biosynthetic processes (protein synthetic -- RNA synthesis and proliferative activity -- DNA synthesis) in the cells of palatine tonsils and structural changes of their tissues depend on characteristics of pathological process. Microbial penetration into the tonsils with formation of an active focus of chronic infection is traced. PMID- 16710174 TI - [Clinico-anatomic rationale of chondroplastic laterofixation of the vocal cordin the treatment of median laryngostenoses]. AB - Stereotopometry used in morphology for imaging three-dimentional organ structure is applied to study individual changeability and spatial relationships of laryngeal structures in adults depending on the type of constitution and form of the neck. Fifty preparations from male corpses with three types of constitution were studied. An original stereotopometer able to localize 106 points on external and internal laryngeal samples to each of three coordinate planes was introduced. The coordinates of certain points were pooled in variational series for characteristics of which the following parameters were determined: arithmetic middling (X), error in arithmetic middling (m), standard deviation (S), correlation coefficient (r). To characterize a constitution type and form of the neck, the following measures were used: length of the body, acromial diameter, chest perimeter, anterior length of the neck, cervical circumference. Morphometric data obtained were investigated with paired correlation analysis. The studies show a definite correlation between spatial relation of laryngeal structures and constitutional type and shape of the neck. This is essential in choice of reconstructive laryngoplasty regarding anthropometric data. PMID- 16710175 TI - [Vertigo and peripheral ischemic cochleovestibular syndrome caused by circulatory insufficiency in the vertebrobasilar system]. AB - We studied characteristics of vestibular and acoustic disorders in patients with arterial hypertension, atherosclerosis and vascular dystonia; correlation between peripheral cochleovestibular syndromes (PCVS) and circulation in the major and intracranial arteries, central hemodynamics, organic changes in the brain. We discovered that anomalies and asymmetry in vertebral arteries diameters, stenoses, reduced stroke and minute blood volumes play an essential role in development of PCVS. The latter are rarely accompanied with ischemic foci in the brain. Medication of vascular vertigo consists in administration of an adequate drug. We believe that betaserk is most effective. PMID- 16710176 TI - [Therapeutic algorithm in stenosis of the larynx and cervical trachea of various etiology]. AB - An algorithm of the treatment of laryngo- tracheal stenosis (LTS) is proposed to raise efficacy of surgical treatment of LTS and to reduce the number of postoperative complications. The algorithm comprises adequate preoperative preparation of the patients including pharmacological prophylaxis of inflammation in the operative zone; effective use of laryngotracheoplasty; prophylaxis of postoperative scarring including reconstruction of created laryngotracheal passage and administration of medicines influencing general and local immunity, tissue metabolism, repair in the region of postoperative inflammation. The efficacy of the algorithm was followed up un 54 (33.7%) patients with laryngostenosis and 106 (66.3%) patients with laryngo-tracheal stenosis. The algorithm application help rehabilitate all the patients with laryngeal stenosis and 92 (86.8%) patients with laryngo-tracheal stenosis. PMID- 16710177 TI - [Most frequent errors in diagnosis and treatment of vocal dysfunction]. AB - Vocal disturbances can be a symptom as laryngeal diseases, and a primary clinical manifestation of an endocrine, neurological or systemic disease, tumors located in the base of the skull, neck, chest. Main causes of the misdiagnosis of vocal disturbances lie in misinterpretation of the results of acoustic study of voice, incomplete case history and inadequately made laryngoscopy. Efficacy of vocal disturbance treatment depends on staged, etiopathogenetic approach with consideration of individual features of the patient. Patients with recurrent and lingering laryngeal diseases should be referred to specialized clinics. PMID- 16710178 TI - [Role of echography in diagnosis of diseases of cervical lymph nodes]. AB - We studied echograms of lymph nodes and clinically similar tumors of the neck in 194 patients. A complex of echographic signs providing the best diagnostic indicators: a minimal size of the node, volume, maximal to minimal size ratio and form. Accuracy of differential diagnosis of benign and malignant lymphadenopathies was 80.4%, sensitivity -- 94.3%, specificity -- 69.4%. In duration of the disease less than 1 month use of computer processing of echograms raised diagnostic accuracy by 9.5%. PMID- 16710179 TI - [The activity of chronic inflammation of palatine tonsils mucosa in chronic tonsillitis before and after treatment using UZOL-01-Ch device]. AB - A low-frequency ultrasound device UZOL-01-Ch was applied for therapy of chronic tonsillitis (CT). The activity of chronic inflammation was followed up under action of 0.05% dioxidine solution coming through the device UZOL-01-Ch on the tissues of palatine tonsils in CT at a clinical remission phase in 34 CT patients. CT activity degrees were proposed which were defined by the presence and distribution of polymorphonuclear neutrophils in inflammatory cellular infiltrate of palatine tonsil mucosa in CT. These degrees help assessed the results of therapeutic effect of the device UZOL-01-Ch on tonsil tissue in CT. Use of the device provided depression of inflammation activity in tonsil mucosa in CT patients at a clinical remission phase. PMID- 16710180 TI - [Postnasal drip syndrome as a cause of persistent cough]. AB - The authors describe postnasal drip syndrome (PNDS) which is a frequent cause of persistent cough. Etiology and pathogenesis of this disease, approaches to diagnosis and treatment of PNDS associated cough are considered. PMID- 16710181 TI - [Early one-stage plastic repair of tracheal lesions in the treatment of flegmon of the neck and acute contact mediastinitis]. AB - Early plastic repair of severe tracheal defects after previous decannulation of the latter in x-ray confirmed efficacy of aspiration flow system of fat spaces of the neck and mediastinum and stability of the patient's condition. This therapeutic policy stops cross-over contamination of fat spaces of the neck, mediastinum, tracheobronchial tree; reduces cost of therapy; facilitates cleansing of foci of purulent surgical infection; shortens time to reconstruction for soft tissue defect of the neck. PMID- 16710182 TI - [Use of autotransplants and implants in ossiculoplasty]. AB - The paper presents a comparative analysis of efficacy of ossiculoplasty (OP) using different transplants. A total of 202 operations were performed. Prostheses made of the fragments of the auditory ossicles, or a cortical layer of the temporal bone (n=81), of nail plate (n=56), titanium implants (n=65) were applied. A satisfactory result was achieved in 72-87% ossiculoplasties. Functional outcomes of OP did not vary significantly with type and material of the prosthesis. Main causes of poor OP results were fixation or displacement of the prosthesis with fibrous tissue; in case of titanium prostheses -- perforation of the tympanic membrane with prosthesis extrusion. PMID- 16710183 TI - [Erespal in combined therapy of acute rhinosinusitis]. PMID- 16710184 TI - [Clinicofunctional parallels in the treatment of allergic rhinitis with mometasone furoate]. PMID- 16710185 TI - [Otogenic abscess of the temporal brain with a break on brain surface and into the lateral ventricle]. PMID- 16710186 TI - [Cases of late diagnosis of acoustic nerve neurinoma]. PMID- 16710187 TI - [Osteoma spongiosum of the maxillary sinus]. PMID- 16710188 TI - [Sphenochoanal polyp in a 3-year-old child]. PMID- 16710189 TI - [A combined trauma of the larynx and trachea: a case report]. PMID- 16710190 TI - [Effective complex homeopatic outpatient treatment of a child with allergic rhinitis and neutropenia]. PMID- 16710191 TI - [Standardization of otorhinolaryngological care]. PMID- 16710192 TI - [Disputable aspects of the problem of exudative otitis media]. PMID- 16710193 TI - [Comparative analysis of efficacy of endovascular interventions in native vascular bed and grafts in patients with ischemic heart disease after coronary artery bypass surgery]. AB - AIM: Investigation of efficacy of endovascular interventions in native vascular bed and grafts, elucidation of factors, affecting immediate and remote prognosis after endovascular treatment. MATERIAL AND METHODS Coronary stenting was carried out in 212 patients who had previously undergone coronary bypass surgery. Stents were implanted into native vessels and grafts in 116 (native vessels group) and 96 (grafts group) patients, respectively. Frequency of angina recurrence and development of complications were assessed during hospitalization and after 1 year. Coronary angiography was repeated after 1 year in 47 and 36 patients in native vessels and grafts groups, respectively. Multifactorial analysis of predictors of complications and angina recurrence was performed with the use of logistic regression. RESULTS AND CONCLUSION: In grafts group signs of distal embolism were observed in 9 patients (9.4%), shunt thrombosis occurred in 2 of these patients. Risk factors of embolism in grafts group were complicated lesions (type C) and length of stenosis >20 mm. There was 1 non-Q wave myocardial infarction after stenting of native vascular bed (0.8%). Angina recurrence was observed after 1 year in 9 (7.8%) and 26 (27.1%), myocardial infarction developed during 1 year in 2 (1.7%) and 3 (3.1%) patients of native vessels and grafts groups, respectively. Risk factors of recurrence of clinic of ischemic heart disease (IHD) after stenting of grafts were time interval between stenting and bypass surgery >5 years and the use of stents without drug coating. More frequent recurrence of clinic of IHD in patients of grafts group was a consequence of higher level of restenosis in stented segments and more frequent progression of atherosclerosis in previously unaffected segments. PMID- 16710194 TI - [Activity of angiotensin converting enzyme in blood serum of patients with stable effort angina]. AB - AIM: To study activity of angiotensin converting enzyme (ACE) in blood serum of patients with stable effort angina. MATERIAL AND METHODS: ACE activity was measured in 11 patients (age 35-45, mean 46.5+/-1.6 years) with typical class II III effort angina without hypertension or contractile insufficiency of the myocardium previously not treated with ACE inhibitors. Diagnosis was verified by angiography (7 patients), data of 24 hour ECG monitoring and exercise tests. Control group comprised 13 practically healthy subjects (age 36-54, mean 48.3+/ 2.09 years). RESULTS: ACE activity in controls was 32.5-57 micromol/min/l (mean 46.2+/-2.0 micromol/min/l), in patients -- 48-110 micromol/min/l (mean 73.5+/-5.9 micromol/min/l), p<0.001. More than 2/3 of patients (7 of 11) had ACE activity above upper limit of norm. PMID- 16710195 TI - [Spectral parameters of heart rate variability and frequency of detection of autoantibodies to beta(1)-adrenoreceptors in patients with tachyarrhythmias: idiopathic and at the background of primary myocardial diseases]. AB - In order to assess parameters of heart rate variability (HRV) and prevalence of autoantibodies against the beta(1)-adrenoreceptors in patients with cardiac arrhythmias we studied 42 patients with arrhythmias and 20 healthy control subjects. Thirty one patients with idiopathic arrhythmias were included in group I: with paroxysmal atrial fibrillation or flutter (n=13), paroxysmal atrial tachycardia (n=2) and paroxysmal ventricular tachycardia (n=16). Group II was formed of 11 patients with paroxysmal ventricular tachycardia and dilated cardiomyopathy or chronic myocarditis. ab1-AR were determined in blood serum by direct immunoassay. Synthetic fragment containing 26 amino acids of ab1-AR second loop was used as antigen. Groups I (54.8%) and II (63.6%) showed similar prevalence of ab1-AR, which was significantly higher than in control subjects (10%) (p<0.005). HRV parameters in I group were lower in ab1-AR-positive compared with ab1-AR-negative patients. At the same time HRV parameters in ab1-AR-positive patients were significantly different from those in controls (p<0.05). In group II HRV parameters of ab1-AR-positive and ab1-AR-negative patients were significantly lower than in control subjects (p<0.05). We suppose, that ab1-AR could participate in dysfunction of chronotropic heart regulation and contribute to development of arrhythmias in patients with structurally normal hearts. PMID- 16710196 TI - [Complex analysis of risk factors of hypertension in subjects involved in managerial activity]. AB - We performed complex analysis of genetic and nongenetic factors facilitating development of arterial hypertension in subjects involved in managerial activity and revealed peculiarities of dermatoglyphic phenotype of susceptibility to this type of pathology. By means of methods of pattern recognition we assessed power and character of effect of each of investigated risk factors. Using mathematical model we elaborated prognosis of efficacy of various preventive procedures. Basing on results of this study we created system of criteria for formation of a group of elevated risk of hypertension and system of measures aimed at prevention of cardiovascular diseases in subjects involved in neuro-emotional labor. PMID- 16710197 TI - [Clinical value of detection of insulin resistance in woman with metabolic syndrome]. AB - Hyperinsulinemic and euglycemic clamp studies were done in 17 women selected from 64 postmenopausal female patients aged 46-67 years with clinical picture of metabolic syndrome, and in 7 control women. Clamp test made it possible to follow dynamics of insulin and other hormones in the process of regulation of glucose level. This method allowed to prove quantitatively the presence of resistance to insulin and hormonal disintegration in patients with metabolic syndrome. Elevation of activity of sympathoadrenal system plays an important role in the pathogenesis of hypertension in metabolic syndrome at the account of augmented sensitivity of receptor apparatus. Lowering of somatotropic hormone level in patients with metabolic syndrome is associated with disturbed somatotropic function of the hypophysis which regulates blood pressure. PMID- 16710198 TI - [Hyperhomocysteinemia in men of Novosibirsk]. AB - Prevalence of hyperhomocysteinemia (HHC) in a sample of male population (n=84, age 50-64 year) of Novosibirsk assessed in the framework of the international project "Determinants of cardio-vascular diseases in the Eastern Europe: multicentral cohort research" during winter-spring period of vitamin deficiency was 50%. In 90.5% of cases HHC was moderate (15-30 micromol/l) and in 9.5% of cases -- medium (30-100 micromol/l). No correlations or independent associations were found between homocysteine blood level and CHD, as well as main risk factors (hyperlipidemia, hypertension, smoking and excessive body weight). No cases of CHD were registered among men with medium HHC. Homocysteinemia correlated positively with age, history of stroke, and negatively -- with alpha-tocopherol concentration in LDL. Men with medium HHC compared with those with normohomocysteinemia had higher systolic and diastolic blood pressure and 29% lower alpha-tocopherol concentration in LDL. PMID- 16710200 TI - [Reversible inhibitory action of adenosine diphosphate ribose on isolated rat heart and rat heart in vivo]. AB - Cardiotropic effects of endogenous regulator of intracellular ionic balance adenosine diphosphate ribose (ADP-ribose) was studied on isolated rat heart using standard retrograde perfusion and on rat heart in vivo by means of implanted arterial catheters. Parameters registered were heart rate, pressure developed in left ventricle of isolated heart and blood pressure of rats in vivo. It was shown that ADP-ribose exerted cardiotropic action on isolated rat heart and that its effects did not depend on effects of its metabolite AMP. In concentrations 10-70 microM ADP-ribose caused negative dose dependent chronotropic effect. Amplitude of contraction of isolated heart changed equivocally. In concentration 70 microM ADP-ribose in 50% of cases produced biphasic action: negative inotropic effect was preceded by positive inotropic effect. In other cases this concentration exerted negative inotropic effect. In concentration 10 microM only negative inotropic effect was observed. Contrary to ADP-ribose its metabolite AMP exerted only monophasic positive inotropic effect on isolated heart. The preparations studied differed in dynamics of cardiotropic action. In experiments in vivo ADP ribose and AMP caused monophasic negative chronotropic and monophasic positive inotropic effects. PMID- 16710199 TI - [Controlled reperfusion improves the metabolic and functional recovery of the isolated heart in rats after total ischemia]. AB - Metabolic and functional effects of a hypocalcium reperfusion solution (RS) with low oxygen content containing d-glucose, trisamine, d-mannitol and I-aspartic acid have been studied in isolated rat hearts. The hearts were initially perfused for 20 min with the Krebs solution under constant left atrial filling pressure of 15 mm Hg and aortic perfusion pressure of 60 mm Hg. Then they were subjected to 30-min total normothermic ischemia followed by 30-min of reperfusion. The Krebs solution (control, n=16) or RS (n=11) were infused in retrograde mode with a rate of 4 ml/min during first 5 min of reperfusion. After that the hearts of both groups were reperfused in antegrade mode with the Krebs solution for 25 min under initial conditions. Short-term infusion of RS markedly improved postischemic functional recovery of cardiac function. After 30-min reperfusion coronary flow, an index of contractile function intensity, expressed as the left ventricle developed pressure-heart rate product, and cardiac work, calculated as the minute volume-aortic perfusion pressure product, recovered up to 92+/-1%, 77+/-1% and 61+/-1% of baseline values, respectively. In the control group the same indices were significantly lower and were 74+/-3%, 48+/-5% and 33+/-2%, respectively (p<0.001). At the end of reperfusion hearts treated with RS compared with the control hearts showed higher myocardial levels of ATP, phosphocreatine (PCr) and total creatine (SCr). These metabolic findings indicate better recovery of energy state and lesser sarcolemmal damage of postischemic cardiomyocytes after RS infusion. Thus, optimization of administration mode and composition of reperfusion solutions is a promising tool to attenuate functional and metabolic disturbances of the postischemic heart. PMID- 16710201 TI - [Disturbances of thyroid function after coronary angiography]. AB - Frequency of thyroid gland functional disturbances after introduction of iodine containing radiographic contrast agents during coronary angiography was studied in 146 patients, residents of the zone of borderline iodine deficit. Frequency of thyroid pathology was high at baseline (39%). Functional state of the thyroid gland was assessed before and 1, 3, 6, 12 months after investigation. During first month after coronary angiography there were several new cases of thyrotoxicosis and hypothyroidism, as well as deterioration of preexisting functional disturbances of the thyroid. Patients with baseline pathology and functional disturbances of thyroid gland before conduction of coronary angiography should be included into risk group of development of iodine induced states. PMID- 16710202 TI - [Singnificance of sex of patients for mortality during 28 days after first myocardial infarction: relation to the presence of Q-wave]. PMID- 16710203 TI - [Dynamics of acute myocardial infarction morbidity and risk of acute ischemic syndrome among population of Kaunas in 1986-2001]. PMID- 16710204 TI - [Neurohumoral mechanisms of pathogenesis of metabolic syndrome]. PMID- 16710207 TI - [Self-monitoring of the effect from indirect anticoagulant agents allows for the decrease of overall mortality, incidence of thromboembolic and hemorrhagic complications]. PMID- 16710208 TI - [Pulmonary hypertension and right ventricular failure. Part III. Patients with congenital heart disease]. AB - In a series of articles the authors discuss literature data concerning epidemiology of pulmonary hypertension, its modern classification, peculiarities of its pathogenesis and treatment in various diseases and conditions. In the third lecture they consider peculiarities of pathogenesis of pulmonary hypertension and right ventricular failure in patients with congenital heart defects, and present literature data on drug treatment and other curative interventions in pulmonary hypertension in patients with Eisenmenger syndrome. PMID- 16710209 TI - [Clinical efficacy and safety of administration of loop diuretic torasemide]. PMID- 16710210 TI - [Mechanism of action and clinical use of ticlopidine]. AB - The literature review contains date on mechanisms of action, pharmacokinetics of ticlopidine, its antithrombotic efficacy in various clinical conditions (ischemic heart and brain diseases, diabetic microangiopathy, occlusive disease of arteries of lower extremities) as well as main adverse effects. PMID- 16710211 TI - [Contemporary approaches to the treatment of arterial hypertension and ischemic heart disease]. AB - Prognostic advantage of a combination of calcium antagonist and angiotensin converting inhibitor (ACEI) over combination of beta-adrenoblocker with thiazide diuretic was proved in a large randomized study of long term treatment of arterial hypertension. In another randomized study administration of ACEI for 12 months caused substantial retardation of cardiac remodeling in elderly patients with normal left ventricular function in postinfarction period. These results are important for a practical physicians as they concern treatment of wide circle of in- out-patients. PMID- 16710212 TI - [Arterial hypertension in the elderly and the status of higher mental functions]. PMID- 16710213 TI - [Current views about the role of purin metabolism disorders in patients with arterial hypertension and losartan potentialities]. AB - The analysis of correlation between cardiovascular diseases and hyperuricaemia as a component of methabolic syndrome has been performed; the emphasis has been done on predominant mechanism of its development -- reduction of renal clearance. The need for primary prophylaxis of stroke with a help of safe metabolically antihypertensive therapy has been substantiated. The results of secondary analysis of LIFE study represented its results on effective primary prophylaxis of stroke depending on the dynamic of serum uric acid levels taking into account uricosuric effect of losartan have been discussed. PMID- 16710214 TI - [Contemporary approaches to the treatment of chronic heart failure in adults (after materials of recommendations of American College of Cardiology and American Heart Association, 2005)]. AB - Detailed and critical analysis of a novel version of the "Updated Guideline for the Diagnosis and Management of Chronic Heart Failure (CHF) in the Adult" prepared by experts of American College of Cardiology and American Heart Association is given. The novel version contains somewhat modified recommendations on the management of patients with CHF. In particular this relates to the place of various classes of drugs in the treatment of CHF due to systolic left ventricular dysfunction. For the first time recommendations on the treatment of patients with CHF and normal left ventricular ejection fraction are presented in detail. Among statements of the updated guideline the following are considered controversial or deserving special discussion. Perindopril is mentioned among recommended ACE inhibitors despite the fact that it has never been studied in long term trials. Results of SENIORS trial are ignored and nebivolol is not included in the number of beta-blockers with proven efficacy. Despite multiple proofs of beneficial effects of aldosterone receptor blockers on clinical course of CHF wide use of spironolactone and eplerenone is not recommended because of multiple communications about life threatening hyperkalemia. Inherent dangers of digoxin therapy are disregarded and the use of cardiac glycosides in patients with sinus rhythm is not prohibited. PMID- 16710215 TI - [Albumin surgical glue application during surgical grafting of thoracic aorta]. AB - Primary results of albumin surgical glue application during surgical grafting of thoracic aorta are analyzed. It is demonstrated that the glue is an effective method of hemostasis, permits the surgeon to decrease time of surgery, intra- and postoperative hemorrhage. PMID- 16710216 TI - [Temporary hemostasis in the injuries of great vessels of different location]. AB - Temporary hemostasis is one of the main points in emergency surgery of patients with trauma of major vessels. Folley catheters were used for temporary hemostasis in bleedings from stab-cut wounds with a narrow wound canal. The method of temporary clamping of the abdominal aorta above the visceral branches during profuse abdominal bleedings of unknown etiology is described in detail. PMID- 16710217 TI - [Injury of inferior vena cava]. AB - Results of treatment of 102 patients with injuries of inferior vena cava (IVC) were analyzed. The causes of injury were penetrating stab-incised wounds of abdomen (67.5% patients), close trauma (28.2%), gunshot wounds (4.3%). Zone of confluence of iliac veins was injured in 7% patients, infrarenal and renal segments -- in 51.9%, suprarenal -- in 21.5%, supra- and retrohepatic -- in 19.6% patients. Injury of one wall of vein was revealed in 63.7% patients, both walls - in 14.7%, complete disruption of vessel -- in 7.9%, avulsion or fissure of hepatic veins -- in 13.7%. In 96.2% patients trauma of IVC were associated with injuries of liver (37.2% cases), small intestine (26.4%), stomach (15.6%), pancreas (12.7%), duodenum (10.7%), large intestine (6.8%), and other organs (10.7%). The side suture (83.7%), circular one (5%), ligation of vein (6.2%), grafting (3.8%), and ligation of left hepatic vein (1.3%) were performed. Reconstructive surgery was carried out by general surgeon in 30% patients, and by vascular surgeon -- in 70%. Lethality was 53.9% (55 patients). Lethality after injuries of supra- and retrohepatic segments of IVC was 100%, suprarenal and renal segments -- 60.6%, infrarenal segment -- 30.6%. PMID- 16710218 TI - [Postoperative electrostimulation in abdominal surgery]. AB - A new method of electric stimulation of the gastrointestinal tract in surgical patients is analyzed. The study group consisted of 201 patients after different abdominal surgeries who had undergone electrostimulation procedures in postoperative period. Conventional methods of paresis prophylaxis and treatment were used in 419 similar patients (control group). Electrostimulation was performed during 20-30 min two times a day from the first day after surgery till complete resolution of paresis. It is concluded that direct electrostimulation in postoperative period reduces duration of paresis, number of postoperative pneumonias ( 6-fold and more) and other complications. Postoperative lethality was 3.98% in the study group and 7.87% in the control group. PMID- 16710220 TI - [Hernioplasty in the treatment of complex and recurrent inguinal hernias]. AB - 20-year (1984-2004) experience in surgical treatment of recurrent and complex hernias in 220 patients after reconstruction of the posterior wall of inguinal canal with sewn-in filamentous frame is analyzed. Results of this treatment are compared with those after standard methods of hernioplasty: Bassini's, Martynov's, Postempsky's, Kukudzhanov's, Bobrov-Girard-Spasokukotsky operations (control group). PMID- 16710219 TI - [Abdominoplasty during operations for ventral hernias]. AB - Anatomic cadavers studies have shown the necessity of one-stage abdominoplasty during herniotomy for ventral hernias. An original method of hernioplasty was developed. It is demonstrated that abdominoplasty in all the cases reduces tension of tissues with maximal effect in defects of the anterior abdominal wall from 32 to 150 cm(2) and subcutaneous fat thickness more than 3 cm. PMID- 16710221 TI - [Endoscopy of bleeding gastroduodenal ulcers]. AB - A new device -- "low-temperature washer" has been developed for better endoscopic visualization of bleeding gastroduodenal ulcers. Endoscopic classification of gastroduodenal bleedings by Forrest has been modified, a new classification of endoscopic hemostasis methods is proposed. Results of diagnosis and treatment of 246 patients are analyzed. Application of the new device increased diagnostic and treatment efficacy of urgent endoscopy, decreased the number of surgeries performed on the peak of the bleeding. PMID- 16710222 TI - [Acute intestinal obstruction after resection of the stomach and pylorus destroying surgeries]. AB - Results of examination of 55 patients with acute intestinal obstruction (AIO) who had undergone resection of the stomach (51) and trunk vagotomy with pyloroplasty (4) were analyzed. Among 29 operated patients AIO were caused by phytobezoars in 21 (72.4%) and comissures in 8 (27.6%) patients. Twenty-two were hospitalized in autumn. Twenty-six patients were treated conservatively, 12 of them were hospitalized in autumn. Small-intestine AIO due to phytobezoars is seen more often in patients with resected stomach. This pathology is characterized by triad of signs: the history of stomach resection, autumn (the season of haw), clinical picture of small-intestine obstruction. PMID- 16710223 TI - [Diagnosis and surgical treatment of liver injuries]. AB - Results of surgical treatment of 214 patients with trauma of the liver are analyzed. The diagnostic value of laparoscopy and urgent dynamic ultrasound investigation is emphasized. Prognostic scale ISS was used for assessment of injury severity. Structure of hepatic trauma and posttraumatic complications is analyzed. The ways of optimization of diagnosis and treatment are determined. PMID- 16710224 TI - [Operations in injuries and strictures of bile ducts]. AB - Analysis of surgical treatment of 336 patients with cicatricial strictures and iatrogenic external fistulas of extrahepatic bile ducts is presented. The causes of these complications were intrasurgical injuries during cholecystectomy (n=302), resection of the stomach (n=21) and echinococcectomy (n=13). In all the cases correction of water-electrolytic balance, treatment and prophylaxis of hepatic and renal insufficiency, control of anaerobic infection were carried out before and after surgery. Restorative operations were performed in 31 (9.2%) patients, reconstructive surgeries -- in 305 (90.7%). Recurrence of the stricture required surgical intervention in 38 (11,3%) patients. Various postoperative complications were diagnosed in 119 (35.4%) patients, 26 (7.7%) patients died. The main causes of death were insufficiency of anastomotic sutures, intoxication, hepatic and renal insufficiency due to purulent cholangitis and cholangiolytic abscesses of the liver. PMID- 16710225 TI - [Recurrence of echinococcosis: pathogenesis, prophylaxis, early diagnosis and treatment]. AB - Complex of measures for prophylaxis of echinococcosis recurrence was developed basing on experimental and clinical studies. Results of surgical treatment of echinococcosis of different location in 983 patients were analyzed. Complex approach to the treatment reduced the rate of recurrences to 0.6% (p=0,0002). Criteria of recovery and algorithm of early diagnosis of echinococcosis recurrence are presented. PMID- 16710226 TI - [Results of combined surgeries for cancer of the colon complicated with stomach and duodenal fistulas]. AB - Positive results of combined surgeries in locally advanced cancer of the colon complicated with gastric fistula (3 patients) and duodenal fistula (1 patient) are presented. Effective treatment of these patients requires complex examination, good preoperative preparation, perfect anesthetic management, adequate operation and intensive postoperative (including antibacterial) therapy. PMID- 16710227 TI - [Aseptic primary-delayed T-type large intestine anastomosis in urgent surgery of the colon]. AB - Experimental studies were carried out on 18 mongrel dogs and 20 cadaver intestinal organ complexes. Possibility of creation of aseptic primary-delayed T type large intestine end-to-side anastomosis with nickel-titan implants is demonstrated. This closed anastomosis creation was used in surgical treatment of 14 patients with obturation large-intestinal tumor obstruction and demonstrated good short-term clinical results. PMID- 16710228 TI - [Course and outcome of surgical infections complicated with sepsis]. AB - A retrospective study of 594 medical histories of patients with surgical infections suggested diagnosis of sepsis in 18.2% cases according to ACCP/SCCM criteria (1992). Pulmonary and abdominal sepsis prevailed and led to higher lethality. Sepsis developed most often as nosocomial infection caused primarily by Acinetobacter spp. and Ps.aeruginosa. Most patients were males aged from 40 to 60 years. The main causes of lethality were septic shock (49.3%) and multiple organ failure syndrome (21%). Time of surgical sanation, antibacterial therapy and its adequacy correlated with the disease outcome. PMID- 16710229 TI - [Wound cover gelepran in the treatment of venous ulcer]. PMID- 16710230 TI - [Pancreaticoduodenal resection for destructive tumor of the pancreatic head with massive bleeding]. PMID- 16710231 TI - [Effective surgical treatment of urethral obstruction]. PMID- 16710232 TI - [Evolution of physical methods of hemostasis in surgery]. PMID- 16710233 TI - [Resection of the stomach with Roux anastomosis]. AB - Results of surgical treatment of patients with gastric ulcer and cancer of the stomach using resection of the stomach with Roux technique were analyzed. Acid producing and evacuatory functions of the stomach were assessed in different periods after surgery. Dynamic scintigraphy provided evaluation of indices of duodenogastric reflux and state of gastric mucous barrier after various types of surgeries. Quality of life was studied late after surgery with original questionnaire. It is concluded that Roux resection of the stomach is not inferior to Bilrot-I resection by functional results, but it has advantages by quality life and rate of operated stomach diseases compared with Bilrot-I and Bilrot-II techniques. PMID- 16710234 TI - [Minimally-invasive surgical treatment of perforated gastroduodenal ulcers]. AB - The results of treatment of 249 patients aged 15 to 78 years with perforated gastroduodenal ulcers were analyzed. The study group consisted of 128 patients in whom video-laparoscopic procedure was used for suturing of ulcer, control group - of 107 patients treated conventionally. Conversion of laparoscopic procedure to open surgery was in 14 cases. It is demonstrated that the intensity of pain syndrome, hospital stay and the number of postoperative complications in the study group were less that in the control group. Indications and contraindications for video-laparoscopic method in these patients were formulated. PMID- 16710235 TI - [Reconstructive jejuno-gastroplasty in the treatment of operated stomach disease]. AB - The results of treatment of 205 patients with operated stomach disease were analyzed. Conservative therapy was effective in 71.7% patients. Surgical treatment was used in 58 (28.3%) patients. Reoperation was performed less than a year after primary surgery in 5.2% patients, 1 to 3 years later -- in 41.4%, 3 to 5 years -- in 36.2%, more than 5 years later - in 17.2% patients. Early after operation complications were observed in 24.1% patients. Clinical and device examinations demonstrate that surgical reconstruction significantly reduces intensity of pathological symptoms in all the patients. PMID- 16710236 TI - [Feasibility of correction of insulin insufficiency after autotransplantation of splenic tissue in duodenopancreatectomy]. AB - Autotransplantation of splenic tissue in two patients after total duodenopancreatectomy and splenectomy for trauma of the duodenum and pancreas has demonstrated positive results and stable compensation of insulin insufficiency in two cases. Experimental studies of autotransplantation of splenic tissue carried out on 20 cats and 100 rats with pancreatectomy- and alloxan-induced diabetes demonstrated good results too. This method may be regarded as the variant of repair therapy with stromal stem-cells of the spleen. PMID- 16710237 TI - [Laparoscopic operation for Hirschsprung's disease in children]. AB - Experience in the treatment of 5 children aged 6 months to 14 years with different types of aganglionosis of the colon using laparoscopic endorectal bringing down of the colon (like Coabe surgery) is analyzed. In all the cases this method permitted mobilization and resection of the intestine (to 60 cm), bringing down intact intestine on the perineum with simultaneous coloanal anastomosis. Duration of the surgery ranged from 120 to 240 min, postoperative hospital stay did not exceed 8 days. Functional and cosmetic results were excellent. The method is characterized by combination of advantages of classical Coabe operation with minimal surgical trauma of laparoscopic mobilization of the colon. PMID- 16710239 TI - [Efficacy of spiral computed tomography in diagnosis of thorax injuries due to combined trauma]. AB - Spiral computed tomography of the thorax was used in 112 patients with mechanical trauma. High efficacy of this method in diagnosis of fracture of posterior and lateral costal parts (69.6%), pneumo- and hydrothorax (100%), traumatic infiltration and gas-filled cysts of the lung (100%), pulmonary hematoma (90%) was demonstrated. Localization of bone fragments in pleural cavity (100%) had special significance. It is demonstrated that the method has low efficacy in the diagnosis of injuries of anterior costal part and sternum (49.3 and 20%). Indications for this method were formulated. The results of the study recommend spiral computed tomography as a priority diagnostic method in combined trauma of the chest. PMID- 16710238 TI - [Endotoxin aggression in the development of hemostatic disorders in children with surgical diseases]. AB - The role of endotoxin aggression in the development of hemostatic disorders was analyzed in 62 children who had undergone urgent and elective surgeries. It is demonstrated that indices of endotoxin-antiendotoxin system and hemostasis before surgery may be regarded as prognostic criteria for development of complications during and after surgery. PMID- 16710240 TI - [Choice of the method of surgical treatment of spleen disruption in combined and isolated abdominal trauma in terms of endosurgery]. AB - The results of the treatment of 378 patients with severe combined (n=270) and isolated (n=108) close abdominal trauma with an injury of the spleen are analyzed. Overall lethality was 11.9% including 7.41% during the first 24 hours. Spleen-preserving surgery was performed in 150 patients, 98% of them had severe injuries of this organ. Splenectomy was performed in 228 patients. Ligation of splenic artery is the indispensable condition of spleen-saving surgery. The level of ligation was validated by the data of own 102 x-ray and 73 anatomic examinations. Optimal level of ligation is located in proximal and middle part of the splenic artery. Spleen-preserving surgeries (especially laparoscopic procedures) significantly reduced lethality, number of postoperative complications and hospital stay. PMID- 16710242 TI - [Surgical treatment of inguinal hernias]. AB - Experience with 1633 surgeries performed in 1592 patients with inguinal hernia is analyzed. In the first period (1979-1987) choice of plastic surgery was empirical, recurrences were diagnosed in 19.6% of examined patients. In the second period (1988-1993) anterior wall was strengthened in simple inguinal hernias, posterior wall -- in difficult, recurrences were seen in 4.9%. In the third period (1994-2000) strengthening of the posterior wall with fascial aponeurotic methods was performed in all types of hernias. Choice of the method depended on height of inguinal space. Recurrences were diagnosed in 0.5% of examined patients. PMID- 16710241 TI - [Surgical rehabilitation of patients with intestinal stoma]. AB - The results of treatment of 145 patients with different type of intestinal stoma created due to urgent abdominal diseases were analyzed. The analysis of rate and nature of postoperative complications in the control group (67 patients) revealed risk factors of postoperative complications and suggested prophylactic measures during all stages of stoma creation and closure. Individual surgical policy reduced number of complications from 34.3 to 12.8%. PMID- 16710243 TI - [Argon plasma flowing the complex treatment of different forms of erysipelas]. AB - The results of complex treatment of 113 patients with different forms of erysipelas were analyzed. In 73 (64.6%) of them argon plasma using "Argon scalpel SP-CPT" in therapeutic regimen was applied as daily local exposure. In 40 (35,4%) patients with complicated form of erysipelas regimen "coagulation" was applied for additional sanitation of wound surface after surgical treatment of purulent focus. The results of treatment in the study group were better than in the control group. Argon plasma may be regarded as effective method of prophylaxis of necrotic and purulent complications particularly in patients with high risk of these complications. PMID- 16710244 TI - [Efficacy of halidor treatment for chronic ischemia of the lower extremities]. PMID- 16710245 TI - [Surgical treatment of leiomyosarcoma of the pulmonary artery under artificial circulation]. PMID- 16710246 TI - [Atypical course of hemorrhoidal thrombosis]. PMID- 16710247 TI - [Use of infected cadaveric donors for liver transplantation]. PMID- 16710248 TI - [Immediate and remote results of stenting of left coronary artery trunk in patients with ischemic heart disease]. AB - The aim of this investigation was assessment of efficacy of stenting in patients with ischemic heart disease with lesions of left coronary artery (LCA) trunk. In the A.N. Bakulev Scientific Center of Cardiovascular Surgery between June 1997 and March 2005 stenting of LCA trunk was carried out in 50 patients (33 with stable effort angina and 17 with acute coronary syndrome). Immediate success rate was 100% in patients with stable angina. In a group of patients with acute coronary syndrome angiographic success rate was 100%. Total lethality in this group was (3 cases) 17.7%. In remote period (6 to 60 months) 33 of 39 patients were examined and recurrence of angina was noted in 7 of them (21.1%). Control angiography was carried out in 16 patients and restenosis of LCA was revealed in 18.75% of cases. The authors believe that stenting of LCA trunk is an effective and safe method of treatment of patients with stable angina and sufficiently safe method in patients with acute coronary syndrome. Roentgenoendovascular treatment may serve as an alternative to aortocoronary bypass surgery especially in isolated lesions of LCA trunk. Application of stents with drug coating allows to cardinally improve long term results of stenting. PMID- 16710249 TI - [Causes and outcomes of coronary arteries perforations]. AB - Coronary artery perforation is a rare but serious complication of percutaneous coronary interventions. Aim of this study--to assess inhospital and long term outcomes in patients in whom perforation occurred during coronary intervention and elucidation of predictors of coronary artery perforation. Between May 1997 and October 2002 perforations were formed in the course of percutaneous interventions in 127 patients what amounted 1.08% of 11,793 patients, subjected to coronary interventions, and 0.77% of 16,494 treated coronary segments. Causes of perforations were complex stenoses, chronic occlusions, calcified lesions, small predicted and minimal vessel lumen, high percent stenosis, use of excimer laser or thromboextrator. Rates of arterial perforations and subsequent adverse events including cardiac tamponade and urgent coronary artery bypass surgery as well as mortality had been declining throughout observation period. PMID- 16710250 TI - [Problems of adaptation in clinical cardiology (quantitative evaluation of adaptation reserve based on data of preconditioning)]. PMID- 16710251 TI - [Cytoarchitectonics and parameters of aggregation of erythrocytes in patients with acute myocardial infarction]. AB - AIM: To assess cytoarchitectonics and parameters of aggregation of erythrocytes in patients with acute myocardial infarction, to elucidate their relationships with risk factors of ischemic heart disease. MATERIAL AND METHODS: We examined 80 patients with acute myocardial infarction, 20 patients with stable angina pectoris comprised control group. Patients were divided into subgroups according to depth of damage of the myocardium and presence of statins in the treatment scheme. Examination included assessment of cytoarchitectonics and parameters of aggregation of erythrocytes, determination of blood plasma lipid spectrum and concentration of fibrinogen. RESULTS AND CONCLUSIONS: Pronounced pathological changes of cytoarchitectonics of erythrocytes and parameters of their aggregation develop in acute myocardial infarction. These changes increase with increase of depth of myocardial damage. Inclusion of statins in the scheme of treatment leads to significant improvement of the parameters studied. This improvement is more pronounced in patients with non-Q-wave myocardial infarction. Presence of significant interrelationships between cytoarchitectonics of erythrocytes and their aggregation and levels of low density lipoprotein cholesterol, concentration of fibrinogen. PMID- 16710252 TI - [Effect of twelve-month combined therapy with perindopril and indapamide on the level of blood pressure and left ventricular hypertrophy in patients with hypertensive disease]. AB - Left ventricular hypertrophy (LVH) in patients with hypertensive disease is associated with unfavorable prognosis. Long term and effective antihypertensive therapy is capable to cause reverse development of LVH. We included in this study 72 patients (27 men, 45 women, age from 34 to 72 years) with untreated 1st and 2nd degree arterial hypertension (systolic blood pressure 140-179 mm Hg and/or diastolic blood pressure 99-109 mm Hg) and echocardiographical signs of LVH (left ventricular myocardial mass index >120 g/m2 in men and >100 g/m2 in women). After that the patients were randomized into 2 groups: the study group (39 patients, mean age 53.0+/-11.6 years) received combination of perindopril and indapamide in initial dose of 2 mg/0.625 mg, and comparison group (33 patients, mean age 54.4+/ 8.2 years) received monotherapy with enalapril (10 mg). Once daily dosing of the preparation provided high level of compliance of patients with treatment. During 12-month therapy with combination of perindopril and indapamide target blood pressure (<140/90) was achieved in 74.4% of patients, during monotherapy with enalapril--in 27.3% of patients. Significant decrease of left ventricular myocardial mass index (LVMMI) with combination therapy was observed by 6th month of treatment (from 260 to 234 g), with monotherapy within same period of time- from 267 to 260 g. As a result of 12 months therapy of patients with I-II degree of hypertension with perindopril and indapamide LVMMI decreased by 17.5% while monotherapy with enalapril was associated with 5.6% decrease of LVMMI. Lowering of LVMMI occurred mainly at the account of decrease of left ventricular wall thickness. PMID- 16710253 TI - [Advantages of long-term controlled stepwise therapy of arterial hypertension with the use of angiotensin converting enzyme inhibitor spirapril]. AB - AIM: Achievement of long-term effective control of blood pressure (BP) in patients suffering from arterial hypertension by means of elaboration of stage by stage program of antihypertensive therapy and proof of its advantage over prescription of hypotensive drugs in real clinical practice. MATERIAL AND METHODS: Effect of spirapril on parameters of vasomotor function of the endothelium were assessed. In randomized comparative prospective study with 1 year duration we included patients with mild and moderate arterial hypertension. The study group received stepwise antihypertensive therapy based on angiotensin converting enzyme inhibitor spirapril. Comparison group continued to take hypotensive drugs prescribed in conditions of outpatients clinic. RESULTS: Sixty patients (79.6% women) were included in the study (30--in the study group, 30--in comparison group). Lowering of BP was observed in the process of therapy in both groups. Among patients of spirapril group lowering of BP was more pronounced and occurred more quickly with achievement of target level of systolic BP <140 mm Hg and diastolic BP <80 1 month after beginning of treatment. Under the influence of therapy with spirapril for 6 months improvement of function of the endothelium was noted. CONCLUSION: In the process of stepwise therapy with the use of spirapril effective long-term control of BP is more quickly achieved compared with standard treatment of arterial hypertension, and improvement of vasomotor function of endothelium also occurs. PMID- 16710254 TI - [The study of organoprotective properties of enalapril in the treatment of arterial hypertension persisting after delivery]. PMID- 16710255 TI - [Levels of fatty acid binding protein before and after direct current cardioversion of atrial fibrillation or flutter in patients without acute coronary syndrome]. AB - It is not known whether FABP suggested as an early marker of myocardial necrosis increases after direct current cardioversion (DCC). Moreover there are no data on FABP levels in patients with atrial fibrillation (AF) and flutter. AIM: To find out whether DCC induces acute changes of FABP levels in patients with AF or flutter without acute coronary syndrome. METHODS: Serum FABP and troponin I (TnI) were measured in 63 pts treated with DCC (monophasic shocks) because of AF (n=45, 71.4%) or atrial flutter (n=18, 28.6%). Initial energy was 50J for atrial flutter and 200J for AF. Median total energy was 200J, max 660J. Blood was sampled before DCC and in 30, 60 min, 3, 6 h after first shock. TnI and FABP were measured with single-step sandwich method ("Hytest" Finland). Upper limit of normal range (ULN) for TnI was 0.4 ng/ml (recommended by manufacturer). Serum FABP was measured in healthy volunteers and 95th percentile was used as ULN (4.67 ng/ml). RESULTS: Most pts (n=40, 63.5%) had baseline FABP > ULN (median for all pts--5.55 ng/ml). In 11 pts (17.4%) baseline FABP was > 2 ULN. No relationship was found between baseline FABP and age, sex, duration of arrhythmia, concomitant ECG changes, and presence of overt coronary heart disease or clinical signs of heart failure. Median FABP level increased after DCC (p for trend 0.00014). FABP levels after DCC correlated with total delivered energy. Delivery of > or = 2 shocks compared with 1 shock was associated with higher FABP (median 60 min--9.65 and 5.24 ng/ml, p=0.009; 3 h--12.41 and 5.84 ng/ml, p=0.01, respectively). Median TnI levels were below ULN at each study point. After DCC TnI did not exceed ULN in 61 pts and remained unchanged in 2 pts with baseline TnI above ULN. CONCLUSION: Unexpectedly most pts with AF and atrial flutter had elevated FABP at baseline. After DCC FABP increased in proportion with total delivered energy. Elevations of FABP levels were not associated with rapid increases of TnI so skeletal muscle damage can be a likely cause of elevated FABP. AF, atrial flutter and DCC for these arrhythmias should be considered as sources of false positive results when FABP is used for diagnosis of acute myocardial infarction. PMID- 16710256 TI - [Glycoprotein IIb/IIIa antagonist eptifibatide in acute coronary syndrome without elevation of ST segment]. AB - Clinical effects of glycoprotein (GP) IIb-IIIa antagonist eptifibatide (Integrilin) and its inhibitory effects on platelet aggregation were studied upon administration of eptifibatide for the treatment of acute coronary syndrome (ACS) without ST segment elevation. Eptifibatide was introduced to 25 patients with unstable angina and non-Q-wave myocardial infarction (MI) according to the following scheme: two boluses with 10 min interval at the dose of 180 mg/kg followed by supporting infusion of 2 mg/kg per min for the first 24 hours and of 1.3 mg/kg for the next 48 hours. Comparative group in which GP IIb-IIIa antagonists were not used upon therapeutic treatment also included 25 patients. All patients received standard basic therapy. 11 patients from the control group and 13 patients from the group with administration of eptifibatide underwent coronary angioplasty during in hospital period. Eptifibatide completely inhibited ADP-induced platelet aggregation within the whole infusion period but after that platelet aggregating activity was quickly recovered--for more than 50% within 6 hours, and completely within 12 hours after the end of infusion. Eptifibatide administration in none of patients was accompanied with the development of dangerous side effects. Thrombocytopenia (50,000 platelets per mm3) was registered in one, and minor bleeding events--in 3 patients. The rate of unfavourable outcomes (MI, refractory or recurrent angina) within first 30 days was almost the same in eptifibatide and control group--32% (8 out of 25) and 36% (9 out of 25) respectively. Thus, despite the complete inhibition of platelet aggregation for 72 hours, eptifibatide administration failed to decrease the amount of adverse events upon treatment of patients with ACS without ST segment elevation. PMID- 16710257 TI - [Asymmetric hypertrophy of interventricular septum in ischemic heart disease]. PMID- 16710258 TI - [Antiatherogenic effects of angiotensin converting enzyme inhibitors from positions (point of view) of evidence based medicine. Part I]. AB - In a series of two articles the authors present data from controlled trials in which cardio-, cerebro-, and vasoprotective effects of angiotensin converting enzyme inhibitors (ACEI) were assessed. The first communication deals with results of randomized studies in most of which no objective proof of antiischemic (antianginal) effects of ACEI was obtained. In seven placebo controlled studies effects of ACEI were evaluated by means of quantitative angiography (or intravascular ultrasound) and in none of them favorable effect of long-term therapy with ACEI on progression of coronary atherosclerosis and restenosis rate after balloon coronary angioplasty was revealed. Retardation of progression of carotid artery atherosclerosis under influence of ACEI was noted in 2 of 4 trials. But diagnostic value of noninvasive methods which were used in these trials is considerably lower than that of angiography or intravascular ultrasound. PMID- 16710259 TI - [Serum markers of fibrosis in patients with arterial hypertension]. PMID- 16710260 TI - [Hypertrophy of left ventricular myocardium as modifiable risk factor: novel possibilities of correction]. AB - Hypertrophy of left ventricular myocardium observed in 15-20% of patients with arterial hypertension is an independent factor which elevates considerably risk of complications of hypertension (ischemic heart disease, chronic heart failure, ventricular arrhythmias). Regression of left ventricular hypertrophy (LVH) at the background of hypotensive therapy is associated with additional lowering of cardiovascular risk. This should be taken into consideration in selection of a hypotensive preparation. Angiotensin converting enzyme inhibitors (ACEI) and angiotensin II receptor blockers (ARB) are believed to have most pronounced ability to cause reverse development of LVH. It has been shown in recently finished PIXEL trial that administration of fixed combination of ACEI perindopril and diuretic indapamide reduced LVH more effectively than monotherapy with high doses of ACEI enalapril and provided better blood pressure control. Therefore in a patient with hypertension and LVH it is expedient to consider combined therapy with ACEI and diuretic including use of their fixed combinations as treatment of choice. PMID- 16710261 TI - [Intensive diabetes treatment and cardiovascular disease in patients with type 1 diabetes. Results of the DCCT/EDIC study]. PMID- 16710262 TI - [Implantable cardioverter defibrillator does not eliminate need for antiarrhythmic drugs in secondary prevention of life threatening ventricular arrhythmias. Results of OPTIC trial]. PMID- 16710263 TI - [Aspirin in primary prevention of cardiovascular events in women and men (a meta analysis of randomized controlled trials)]. PMID- 16710264 TI - [The use of statins in "old" randomized controlled trials was not associated with either increased or decreased risk of cancer or cancer deaths]. PMID- 16710265 TI - [Pulmonary hypertension and right ventricular failure. Part II. Patients with left ventricular involvement]. AB - In a series of articles the authors discuss literature data concerning epidemiology of pulmonary hypertension, modern classification; peculiarities of its pathogenesis and treatment in various diseases and conditions. In the second part they consider peculiarities of pathogenesis of pulmonary hypertension and right ventricular failure in patients with predominant involvement of left cardiac chambers. Effects of various classes of drugs used in the treatment of left ventricular heart failure including their influence on pulmonary circulation and functional state of the right ventricle are also discussed. PMID- 16710266 TI - [Investigation of efficacy of cardiovascular drugs in women]. PMID- 16710267 TI - [Peculiarities of lipid disorders in patients with type II diabetes mellitus: in which cases we should administer statins?]. AB - Cardiovascular complications are a principal cause of death rate in the patients with type 2 diabetes, that in turn contribute to cardiovascular diseases and complications in adult population. According to UKPDS data high LDL Ch level is strong predictor of CHD development in patients with type 2 diabetes and its 1 mmol/l level increase, due the coronary risk increase on 57%. Patients with type 2 diabetes have the target levels of lipids enough rigid: the total Ch < 4.5 mmol/l, LDL Ch < 2.5 mmol/l, triglycerides < 1.7 mmol/l and HDL Ch > 1 mmol/l. The results of CARDS study have shown, that the treatment of the type 2 diabetes patients with mild hypercholesterolemia without CHD by atorvastatin in a doze of 10 mg during 5 years decreased cardiovascular risk on 37%, an stroke on 48% and the total death rate on 27%. In practice, 89% of the patients with type 2 diabetes comes to light deviations in Ch level. The lipid spectrum assay of should join to the obligatory diagnostic procedures in all type 2 diabetes patients despite of the age and sex. Statin therapy in the diabetes patients is as necessary as the antidiabetic treatment. PMID- 16710268 TI - [Comparative in vitro analysis of physico-mechanical properties of materials used for cervical defects filling]. PMID- 16710269 TI - [Histamine-releasing activity of dental materials employed as root canal filling]. AB - Four samples of dental materials (DM) estd for root canal filling, Septodent (S), Zinkoxide (Z), Foredent (F) and Cialite Regular (CR), were studied for their ability in vitro to release histamine (HR) employing the computerized fiber glass based histamine release assay using whole blood from allergic patients and healthy donors. It was found that F induced high and S low levels of HR from blood basophils from both allergic and non-allergic subjects. Z and CR did not induce HR from blood basophils similar to the negative control. The HR assay may be a useful method for preliminary investigation of DM employed for root canal fillings for safety as well as for selection of specific DM for individual patients. PMID- 16710271 TI - [Distribution of some biologically active substances in human gingiva]. PMID- 16710272 TI - [Influence of mandibular fracture upon intensity and prevailing direction of carbonate and citrate transport between blood and bones]. AB - Using new coefficient of the difference relative radioactivity (CD(RRA)) bone/blood there was studied [(14)C]-carbonate and [3-(14)C]-citrate in 1-month and 6-months rats after right-side fracture of the mandible in prevailing direction between blood and bone. Animals were taken out from the experiment 20 min after intraperitoneal injection of [(14)C]-carbonate and [3-(14)C]-citrate on 7th day after (stage of cell-fibrous callosity), on 14th day after (stage of chondroid callosity) and on 28th day after (stage of the primary bone callosity) the experiment start. Changes after injections of marked citrate and carbonate were similar. Input % (ratio pulse/min in bone and blood serum) and RRA in 1 week increased sharply, decreased in 2 weeks and less changing in 4 weeks. After fracture CD(RRA) in traumatized mandible and non traumatized femur in rats were changing in the opposite directions that testifies to the different prevailing directions of marked substances transport between blood and bones. PMID- 16710273 TI - [Influence of course use of chewing gum on alveolar bone density]. AB - Efficacy of course use of chewing gum for alveolar bone reinforcement was studied with the use of digital X-ray-graphy at three levels of interalveolar bone in 93 patients with clinically different periodontal tissue conditions. It was established that optical density of alveolar bone is higher on the working side than on the inactive side and it depended upon clinical status of periodontal tissues. Top of the interalveolar bone had the lowest density, the middle part - the highest. Course use of chewing gum in the optimal regimen increased interalveolar bone optical density, at that to a greater extent on the inactive side and in the upper part of it. PMID- 16710274 TI - [Comparative study of homeopathic remedies clinical efficacy in comprehensive treatment of inflammatory periodontal diseases in patients with burdened allergic status]. AB - Comparative analysis of clinical efficacy of 3 antihomotoxic homeopathic preparations (Traumeel S, Engistol and Echinacea compositum S) with non-specific immunostimulating and anti-inflammatory effects was performed. The study showed that Traumeel S had maximal anti-inflammatory effect. In proportion as destructive process weighting the efficacy of homeopathic preparations was decreased. The preparations are indicated for comprehensive treatment of inflammatory periodontal diseases in patients with burdened allergic status or heavy concomitant pathology. PMID- 16710275 TI - [Study of local immunostimulating effect of the use of different pharmaceutical forms of the remedy Traumeel S in treatment of inflammatory parodontal diseases]. AB - In order to optimize the way of the remedy "Traumeel S" administration there was studied by the luminol-depending chemiluminescent method immunostimulating effect of its different pharmaceutical forms on functional status of gingival blood granulocytes of patients with inflammatory periodontal diseases. The immunostimulating effect of Traumeel S depended upon only the inflammatory periodontal disease severity and not the ways of its introduction. But the use of the preparation in the pellet form gave some additional advantage: simplicity of use, painlessness, dosage accuracy and possibility not to attend doctor. PMID- 16710276 TI - [Efficacy study of immunomodulator Gepon in treatment of oral membrane diseases in patients with complications in the form of filling material extrusion into mandibular canal]. AB - Experience in new domestic immunomodulator Gepon application in practice of treatment of virus diseases and their manifestations in oral cavity of patients with complications in the form of filling material extrusion into mandibular canal is submitted. In a combination to traditional techniques of the treatment the preparation has shown high efficacy. PMID- 16710277 TI - [Operative treatment tactics in cases of filling material extrusion into mandibular canal]. AB - The way of the surgical help to the patient with endodontic treatment complications in the form of filling material extrusion into mandibular canal is submitted. The technique of operative intervention was developed in experiment and on cadaveric material. PMID- 16710278 TI - [Fungi lesions of maxillary sinus and comparative estimation of chlorhexedine bigluconate and Biopag antimicrobial action]. AB - Having bacteriological assay of the material from maxillary sinuses the authors confirmed the presence of fungus infection. There were picked out and identified fungi Aspergillus niger, Candida albicans and Actinomyces israelii. Aspergillus niger can imitate on roentgenograms the picture of foreign body presence in maxillary sinus. Comparison of bactericidal activity of chlorhexedine bigluconate and Biopag showed the advantages of Biopag preparation. PMID- 16710280 TI - [Method of tongue size reduction with maintenance of taste and other types of sensitivity]. AB - All known operations for tongue size reduction in cases of macroglossia foresee end and part of back tongue excision. In this case patient will get rid of gustatory and other types of sensitivity and it is considered inevitable. Method is described to reduce tongue size allowing preserve all types of sensitivity. During the operation of end tongue excision narrow part of its edge is left in the form of mucous-muscle flap on feeding stem. Scheme of the operation is described. PMID- 16710279 TI - [Hypoxic training -- effective method of raising skin vitality in cases of local plastic operations]. AB - In spite of the complications after expander use the advantages of local plastics predetermine constant return to the method. Systemic use of hypoxic training produces stimulating effect on the skin microcirculation and can be used widely in clinical surgical practice in cases of local plastic operations to prevent possible complications reducing consequences of blood supply disturbance and tissue oxygen provision. The method of hypoxic adaptation let us receive as non medicamentous mean to prepare for shorter period of time more vital flap. PMID- 16710281 TI - [New technologies and ways of endodontic-endoossal implantation improving]. AB - There were developed, tested and used in practice new transdental implants (TDI) considerably widening the indications to endodontic-endoossal implantation. Universal TDI has a number of advantages over the known analogs and meant for osseointegration and fibro-osseointegration. The implant provides full tooth fixation in small bone volume, on maxilla including. PMID- 16710283 TI - [Eru somparative analysis of efficiency of various methods of operative access in cases of complications after filling material extrusion into mandibular channel]. AB - Comparative analysis of efficiency of various methods of operative access to mandibular channel is presented in the framework of the surgical help to the patients with complications endodontic treatment in the form of some filling material extrusion into mandibular channel. PMID- 16710282 TI - [Prosthetic treatment of patients with the use of one-stage intraosseous implants according to the Biomal-implant system]. AB - There was developed and patented new one-stage intraosseous titanium monolithic implant. Detailed lab-clinical analysis of long-term results of 185 patients with the use of intraosseous implants according to the Biomal-implant system was presented. Based on their own clinical experience the authors gave recommendations of shortcut method of prosthetic treatment of patients with different types of adentia. PMID- 16710284 TI - [Frequency-resonance analysis of dental implants stability]. PMID- 16710285 TI - [Possibilities and prospects of the dental apparatus Optodan use for the magnetic laser treatment of stomatologic diseases]. PMID- 16710286 TI - Neonatal arterial hypertension in nephro-urological malformations in a tertiary care hospital. AB - An increase in the survival of neonates with antenatal diagnosis of malformations was achieved by the recent technical advances in neonatal intensive care units. The aim of this article is to describe the experience with neonatal arterial hypertension, in newborns with nephro-urological malformations, in a tertiary care referral Nursery, in a period of 4 years. Newborn medical records from the Nursery Annex to the Maternity of Hospital das Clinicas, School of Medicine, University of Sao Paulo, with the diagnosis of nephro-urological malformations and systemic arterial hypertension (SAH) at hospital discharge, in a period from January 1999 to January 2003, were retrospectively analysed. Among 10.278 live newborns in the studied period, 15 (0.15%) newborns were compatible with our inclusion criteria. Of these 15 newborns, 12 (80%) were male and three were premature (20%). In relation to aetiology, 13 (87%) showed urological malformations, 1 (6%) chronic renal insufficiency secondary to kidney dysplasia and one (6%) autosomal recessive polycystic kidney disease. SAH control was achieved with monotherapy in eight patients (53%), five patients (33%) needed an association of two drugs (calcium-channel blocker and angiotensin converting enzyme (ACE) inhibitor), one child used three types of antihypertensive drugs (calcium-channel blocker, ACE inhibitor and hydrochlorothiazide) for pressoric control and one child's blood pressure (BP) was controlled exclusively by peritoneal dialysis. The incidence of nephro-urological malformations in our service during the studied period was 0.89%. SAH incidence among these newborns was 19%. Our data reinforce previous studies pointing to the necessity to consider children with nephro-urological malformations as a risk group for SAH, who should have the BP evaluated since the neonatal period. PMID- 16710287 TI - Rationale and design of a study to evaluate management of proteinuria in patients at high risk for vascular events: the IMPROVE trial. AB - Declining kidney function predicts increasing cardiovascular risk in people with hypertension. Microalbuminuria is a marker for cardiovascular risk and declining kidney function. Agents that block the renin-angiotensin-aldosterone system (RAAS), notably angiotensin-converting enzyme (ACE) inhibitors and angiotensin receptor blockers (ARBs), reduce proteinuria and microalbuminuria, lower blood pressure and slow the progression of proteinuric kidney disease. Evidence is accumulating that the combination of an ACE inhibitor and an ARB is the optimal means of RAAS blockade in this setting, slowing the progression of nephropathy independently of blood pressure lowering to a greater degree than can be achieved using maximum approved doses of either agent alone. However, the emerging therapeutic potential of ACE inhibitor/ARB combination therapy in hypertensive kidney disease requires further characterization. The Irbesartan in the Management of PROteinuric patients at high risk for Vascular Events trial aims to determine definitively whether the combination therapy of an ARB, irbesartan and an ACE inhibitor, ramipril, is more effective than ramipril alone in reducing the urinary albumin excretion rate in patients at high cardiovascular risk with hypertension and proteinuria or microalbuminuria. PMID- 16710288 TI - Self-monitoring of home blood pressure with estimation of daily salt intake using a new electrical device. AB - We investigated a simple device to monitor daily salt intake at home and examined the relationship between salt excretion and morning blood pressure in order to enable patients to better manage daily salt intake and hypertension. The correlation between 24-h urinary salt excretion and measured value with salt monitor from overnight urine was significant (n = 224, r = 0.72, P < 0.001). A total of 46 volunteers participated for more than 3 weeks by measuring daily salt intake and morning blood pressure. The relationship between predicted daily salt excretion and blood pressure was examined with use of 3-day moving average. Mean salt excretion and systolic blood pressure (SBP) significantly decreased by the end of the trial (i.e., salt excretion decreased from 158+/-31 to 149+/-30 mmol/day and SBP from 137+/-17 to 133+/-16 mm Hg). Of 46 participants, 18 (39%) had a significant correlation between predicted daily salt excretion and blood pressure (r > 0.4, P < 0.05, n > 21), indicating sodium sensitivity. An additional 17% had a positive correlation that did not reach statistical significance (0.2 < r < or = 0.4), and the remaining 44% had no correlation (r < or = 0.2). Mean decrease in blood pressure per decrease in salt (g) (17 mmol) intake in the 18 participants with a significant correlation was 3.3 mm Hg (SBP) and 1.5 mm Hg (diastolic blood pressure), which was higher than that reported for other studies. Hypertensive patients not using medication showed the largest decrease. We conclude that daily monitoring of salt intake and morning blood pressure will be useful for management of hypertension. PMID- 16710289 TI - C-reactive protein and echocardiography have little impact on risk stratification in never-treated hypertensive patients. AB - The aim of this study was to assess the contribution of increased concentrations of high-sensitivity C-reactive protein (hsCRP) and echocardiography to risk stratification according to the 2003 European guidelines for the management of arterial hypertension in patients with untreated hypertension. A total of 207 consecutive medical outpatients with untreated hypertension were included. History and clinical examination, electrocardiography, laboratory analyses including the measurement of hsCRP and echocardiography were performed in all patients. Patients were classified into four risk groups with and without using echocardiography and hsCRP concentrations of at least 10 mg/l according to the 2003 guidelines for the management of hypertension. The majority of the 207 patients (81%) were at moderate or high cardiovascular risk before adding echocardiography and/or hsCRP to the risk stratification process. When echocardiography was included, only three patients were reclassified from the moderate added risk to the high added risk group. Adding hsCRP concentrations of at least 10 mg/l had no impact on risk stratification. Using an hsCRP cutoff level of 3 mg/l, one patient was at moderate instead of low added risk, eight patients were at high instead of moderate added risk and one patient was at very high instead of high added risk. We conclude that hsCRP at the proposed cutoff level of 10 mg/l has no impact on risk stratification in outpatients with untreated hypertension. An hsCRP cutoff level of 3 mg/l may be more suitable for risk stratification. Finally, our data suggest that depending on the population studied, there is minimal impact of echocardiography on risk stratification. PMID- 16710290 TI - Does it matter whether patients take their antihypertensive medication as prescribed? The complex relationship between adherence and blood pressure control. PMID- 16710291 TI - Spontaneous renal artery dissection: three cases and clinical algorithms. AB - Spontaneous renal artery dissection (SRAD) is rare. Clinical manifestations vary from minimal symptoms to life-threatening hypertension. We analysed three cases from our institution and conducted a literature review in order to design diagnostic and treatment algorithms for SRAD. PMID- 16710292 TI - The role of ischaemia and pain in the blood pressure response to exercise stress testing in patients with coronary heart disease. AB - Silent myocardial ischaemia is a common phenomenon in patients with coronary heart disease. However, very little is known about the underlying mechanisms of silent ischaemia. One potential pathway that may contribute to this absence of pain is increased blood pressure. The main aim of the current study was to assess the associations among blood pressure, pain and ischaemia in patients undergoing a standard exercise stress test. We hypothesized that patients who experienced chest pain during exercise would have lower baseline and peak blood pressures compared to those who did not experience chest pain. A total of 1,355 patients (418 women) who underwent a single-photon emission computed tomography treadmill exercise stress test and had not experienced a cardiac event in the past 2 weeks participated in the current study. Myocardial perfusion defects were assessed at rest and during the stress challenge. Systolic blood pressure (SBP), diastolic blood pressure, heart rate (HR) and rate pressure product (RPP) were assessed during rest and at peak exercise. There were no main effects of either pain or ischaemia on the baseline cardiovascular variables. Peak exercise data revealed main effects of pain on SBP, RPP and HR, and main effects of ischaemia on SBP and RPP, controlling for age, sex, baseline level, medication status and cardiac history. These findings suggest that acute rather than chronic increases in blood pressure may be one mechanism to explain the phenomena of silent myocardial ischaemia in cardiac patients, and may potentially provide a target for future treatment strategies. PMID- 16710293 TI - NR2B tyrosine phosphorylation modulates fear learning as well as amygdaloid synaptic plasticity. AB - Phosphorylation of neural proteins in response to a diverse array of external stimuli is one of the main mechanisms underlying dynamic changes in neural circuitry. The NR2B subunit of the NMDA receptor is tyrosine-phosphorylated in the brain, with Tyr-1472 its major phosphorylation site. Here, we generate mice with a knockin mutation of the Tyr-1472 site to phenylalanine (Y1472F) and show that Tyr-1472 phosphorylation is essential for fear learning and amygdaloid synaptic plasticity. The knockin mice show impaired fear-related learning and reduced amygdaloid long-term potentiation. NMDA receptor-mediated CaMKII signaling is impaired in YF/YF mice. Electron microscopic analyses reveal that the Y1472F mutant of the NR2B subunit shows improper localization at synapses in the amygdala. We thus identify Tyr-1472 phosphorylation as a key mediator of fear learning and amygdaloid synaptic plasticity. PMID- 16710294 TI - C-terminal-binding protein directly activates and represses Wnt transcriptional targets in Drosophila. AB - Regulation of Wnt transcriptional targets is thought to occur by a transcriptional switch. In the absence of Wnt signaling, sequence-specific DNA binding proteins of the TCF family repress Wnt target genes. Upon Wnt stimulation, stabilized beta-catenin binds to TCFs, converting them into transcriptional activators. C-terminal-binding protein (CtBP) is a transcriptional corepressor that has been reported to inhibit Wnt signaling by binding to TCFs or by preventing beta-catenin from binding to TCF. Here, we show that CtBP is also required for the activation of some Wnt targets in Drosophila. CtBP is recruited to Wnt-regulated enhancers in a Wnt-dependent manner, where it augments Armadillo (the fly beta-catenin) transcriptional activation. We also found that CtBP is required for repression of a subset of Wnt targets in the absence of Wnt stimulation, but in a manner distinct from previously reported mechanisms. CtBP binds to Wnt-regulated enhancers in a TCF-independent manner and represses target genes in parallel with TCF. Our data indicate dual roles for CtBP as a gene-specific activator and repressor of Wnt target gene transcription. PMID- 16710295 TI - Protonation-mediated structural flexibility in the F conjugation regulatory protein, TraM. AB - TraM is essential for F plasmid-mediated bacterial conjugation, where it binds to the plasmid DNA near the origin of transfer, and recognizes a component of the transmembrane DNA transfer complex, TraD. Here we report the 1.40 A crystal structure of the TraM core tetramer (TraM58-127). TraM58-127 is a compact eight helical bundle, in which the N-terminal helices from each protomer interact to form a central, parallel four-stranded coiled-coil, whereas each C-terminal helix packs in an antiparallel arrangement around the outside of the structure. Four protonated glutamic acid residues (Glu88) are packed in a hydrogen-bonded arrangement within the central four-helix bundle. Mutational and biophysical analyses indicate that this protonated state is in equilibrium with a deprotonated tetrameric form characterized by a lower helical content at physiological pH and temperature. Comparison of TraM to its Glu88 mutants predicted to stabilize the helical structure suggests that the protonated state is the active form for binding TraD in conjugation. PMID- 16710296 TI - UBP43 is a novel regulator of interferon signaling independent of its ISG15 isopeptidase activity. AB - Interferons (IFNs) regulate diverse cellular functions through activation of the Janus kinase-signal transducer and activator of transcription (JAK-STAT) pathway. Lack of Ubp43, an IFN-inducible ISG15 deconjugating enzyme, leads to IFN hypersensitivity in ubp43-/- mice, suggesting an important function of Ubp43 in downregulation of IFN responses. Here, we show that Ubp43 negatively regulates IFN signaling independent of its isopeptidase activity towards ISG15. Ubp43 functions specifically for type I IFN signaling by downregulating the JAK-STAT pathway at the level of the IFN receptor. Using molecular, biochemical, and genetic approaches, we demonstrate that Ubp43 specifically binds to the IFNAR2 receptor subunit and inhibits the activity of receptor-associated JAK1 by blocking the interaction between JAK and the IFN receptor. These data implicate Ubp43 as a novel in vivo inhibitor of signal transduction pathways that are specifically triggered by type I IFN. PMID- 16710297 TI - Crystal structure of CHP2 complexed with NHE1-cytosolic region and an implication for pH regulation. AB - The plasma membrane Na+/H+ exchangers (NHE) require calcineurin B homologous protein (CHP) as an obligatory binding partner for ion transport. Here, we report the first crystal structure of CHP (CHP2 isoform) in complex with its binding domain in NHE1. We show that the cytoplasmic alpha-helix of NHE1 is inserted into the hydrophobic cleft formed by N- and C-lobes of CHP2 and that the size and shape of this crevice together with hydrogen bond formation at multiple positions assure a high degree of specificity for interaction with NHE members. Structure based mutagenesis revealed the importance of hydrophobic interactions between CHP/NHE1 for the function of NHE1. Furthermore, the crystal structure shows the existence of a protruding CHP-unique region, and deletion of this region in CHP2 inhibited the NHE1 activity by inducing the acidic shift of intracellular pH dependence, while preserving interaction with NHE1. These findings suggest that CHP serves as an obligatory subunit that is required both for supporting the basic activity and regulating the pH-sensing of NHE1 via interactions between distinct parts of these proteins. PMID- 16710298 TI - Structure and E3-ligase activity of the Ring-Ring complex of polycomb proteins Bmi1 and Ring1b. AB - Polycomb group proteins Ring1b and Bmi1 (B-cell-specific Moloney murine leukaemia virus integration site 1) are critical components of the chromatin modulating PRC1 complex. Histone H2A ubiquitination by the PRC1 complex strongly depends on the Ring1b protein. Here we show that the E3-ligase activity of Ring1b on histone H2A is enhanced by Bmi1 in vitro. The N-terminal Ring-domains are sufficient for this activity and Ring1a can replace Ring1b. E2 enzymes UbcH5a, b, c or UbcH6 support this activity with varying processivity and selectivity. All four E2s promote autoubiquitination of Ring1b without affecting E3-ligase activity. We solved the crystal structure of the Ring-Ring heterodimeric complex of Ring1b and Bmi1. In the structure the arrangement of the Ring-domains is similar to another H2A E3 ligase, the BRCA1/BARD1 complex, but complex formation depends on an N terminal arm of Ring1b that embraces the Bmi1 Ring-domain. Mutation of a critical residue in the E2/E3 interface shows that catalytic activity resides in Ring1b and not in Bmi1. These data provide a foundation for understanding the critical enzymatic activity at the core of the PRC1 polycomb complex, which is implicated in stem cell maintenance and cancer. PMID- 16710299 TI - Topoisomerase II, not topoisomerase I, is the proficient relaxase of nucleosomal DNA. AB - Eukaryotic topoisomerases I and II efficiently remove helical tension in naked DNA molecules. However, this activity has not been examined in nucleosomal DNA, their natural substrate. Here, we obtained yeast minichromosomes holding DNA under (+) helical tension, and incubated them with topoisomerases. We show that DNA supercoiling density can rise above +0.04 without displacement of the histones and that the typical nucleosome topology is restored upon DNA relaxation. However, in contrast to what is observed in naked DNA, topoisomerase II relaxes nucleosomal DNA much faster than topoisomerase I. The same effect occurs in cell extracts containing physiological dosages of topoisomeraseI and II. Apparently, the DNA strand-rotation mechanism of topoisomerase I does not efficiently relax chromatin, which imposes barriers for DNA twist diffusion. Conversely, the DNA cross-inversion mechanism of topoisomerase II is facilitated in chromatin, which favor the juxtaposition of DNA segments. We conclude that topoisomerase II is the main modulator of DNA topology in chromatin fibers. The nonessential topoisomerase I then assists DNA relaxation where chromatin structure impairs DNA juxtaposition but allows twist diffusion. PMID- 16710300 TI - Sws1 is a conserved regulator of homologous recombination in eukaryotic cells. AB - Rad52-dependent homologous recombination (HR) is regulated by the antirecombinase activities of Srs2 and Rqh1/Sgs1 DNA helicases in fission yeast and budding yeast. Functional analysis of Srs2 in Schizosaccharomyces pombe led us to the discovery of Sws1, a novel HR protein with a SWIM-type Zn finger. Inactivation of Sws1 suppresses the genotoxic sensitivity of srs2Delta and rqh1Delta mutants and rescues the inviability of srs2Delta rqh1Delta cells. Sws1 functions at an early step of recombination in a pro-recombinogenic complex with Rlp1 and Rdl1, two RecA-like proteins that are most closely related to the human Rad51 paralogs XRCC2 and RAD51D, respectively. This finding indicates that the XRCC2-RAD51D complex is conserved in lower eukaryotes. A SWS1 homolog exists in human cells. It associates with RAD51D and ablating its expression reduces the number of RAD51 foci. These studies unveil a conserved pathway for the initiation and control of HR in eukaryotic cells. PMID- 16710302 TI - Molecular basis for temperature sensing by an RNA thermometer. AB - Regulatory RNA elements, like riboswitches, respond to intracellular signals by three-dimensional (3D) conformational changes. RNA thermometers employ a similar strategy to sense temperature changes in the cell and regulate the translational machinery. We present here the first 3D NMR structure of the functional domain of a highly conserved bacterial RNA thermometer containing the ribosome binding site that remains occluded at normal temperatures (30 degrees C). We identified a region adjacent to the Shine-Dalgarno sequence that has a network of weak hydrogen bonds within the RNA helix. With the onset of heat shock at 42 degrees C, destabilisation of the RNA structure initiates at this region and favours the release of the ribosome binding site and of the start codon. Deletion of a highly conserved G residue leads to the formation of a stable regular RNA helix that loses thermosensing ability. Our results indicate that RNA thermometers are able to sense temperature changes without the aid of accessory factors. PMID- 16710301 TI - Modulatory and catalytic modes of ATP binding by the calcium pump. AB - We present crystal structures of the calcium-free E2 state of the sarcoplasmic reticulum Ca2+ -ATPase, stabilized by the inhibitor thapsigargin and the ATP analog AMPPCP. The structures allow us to describe the ATP binding site in a modulatory mode uncoupled from the Asp351 phosphorylation site. The Glu439 side chain interacts with AMPPCP via an Mg2+ ion in accordance with previous Fe2+ cleavage studies implicating this residue in the ATPase cycle and in magnesium binding. Functional data on Ca2+ mediated activation indicate that the crystallized state represents an initial stage of ATP modulated deprotonation of E2, preceding the binding of Ca2+ ions in the membrane from the cytoplasmic side. We propose a mechanism of Ca2+ activation of phosphorylation leading directly from the compact E2-ATP form to the Ca2E1-ATP state. In addition, a role of Glu439 in ATP modulation of other steps of the functional cycle is suggested. PMID- 16710303 TI - The BCL11AXL transcription factor: its distribution in normal and malignant tissues and use as a marker for plasmacytoid dendritic cells. PMID- 16710304 TI - JAK2 mutation in a boy with polycythemia vera, but not in other pediatric hematologic Disorders. PMID- 16710305 TI - Hypoxia suppresses BCR/Abl and selects imatinib-insensitive progenitors within clonal CML populations. PMID- 16710306 TI - Genetic evidence for complex speciation of humans and chimpanzees. AB - The genetic divergence time between two species varies substantially across the genome, conveying important information about the timing and process of speciation. Here we develop a framework for studying this variation and apply it to about 20 million base pairs of aligned sequence from humans, chimpanzees, gorillas and more distantly related primates. Human-chimpanzee genetic divergence varies from less than 84% to more than 147% of the average, a range of more than 4 million years. Our analysis also shows that human-chimpanzee speciation occurred less than 6.3 million years ago and probably more recently, conflicting with some interpretations of ancient fossils. Most strikingly, chromosome X shows an extremely young genetic divergence time, close to the genome minimum along nearly its entire length. These unexpected features would be explained if the human and chimpanzee lineages initially diverged, then later exchanged genes before separating permanently. PMID- 16710307 TI - Multiphoton microscopy for the investigation of dermal penetration of nanoparticle-borne drugs. AB - Multiphoton microscopy (MPM) of a dually fluorescence-labeled model system in excised human skin is employed for high-resolution three-dimensional (3D) visualization in order to study the release, accumulation, and penetration properties of drugs released from nanoscale carrier particles in dermal administration. Polymer particles were covalently labeled with fluorescein, whereas Texas Red as a drug-model was dissolved in the particles to be released to the formulation matrix. Single nanoparticles on skin could easily be localized and imaged with diffraction-limited resolution. The temporal evolution of the fluorescent drug-model concentration in various skin compartments over more than 5 hours was investigated by multiphoton spectral imaging of the same area of the specimen. The 3D penetration profile of the drug model in correlation with skin morphology and particle localization information is obtained by multiple laser line excitation experiments. MPM combined with spectral imaging was found to allow noninvasive long-term studies of particle-borne drug-model penetration into skin with subcellular resolution. By dual color labeling, a clear discrimination between particle-bound and released drug model was possible. The introduced technique was shown to be a powerful tool in revealing the dermal penetration properties and pathways of drugs and nanoscale drug vehicles on microscopic level. PMID- 16710308 TI - Caged-iron chelators a novel approach towards protecting skin cells against UVA induced necrotic cell death. AB - Exposure of human skin cells to solar UVA radiation leads to an immediate dose dependent increase of labile iron that subsequently promotes oxidative damage and necrotic cell death. Strong iron chelators have been shown to suppress cell damage and necrotic cell death by moderating the amount of labile iron pool (LIP), but chronic use would cause severe side effects owing to systemic iron depletion. Prodrugs that become activated in skin cells at physiologically relevant doses of UVA, such as "caged-iron chelators", may provide dose- and context-dependent release. Herein, we describe prototypical iron chelator compounds derived from salicylaldehyde isonicotinoyl hydrazone and pyridoxal isonicotinoyl hydrazone and demonstrate that the intracellular LIP and subsequent necrotic cell death of human skin fibroblasts is significantly decreased upon exposure to a combination of the prototypical compounds and physiologically relevant UVA doses. Iron regulatory protein bandshift and calcein fluorescence assays reveal decreased intracellular LIP following irradiation of caged-chelator treated cells, but not in control samples where either UVA light, or caged chelator is absent. Furthermore, flow cytometry shows that these compounds have no significant toxicity in the skin fibroblasts. This novel light-activated prodrug strategy may therefore be used to protect skin cells against the deleterious effects of sunlight. PMID- 16710309 TI - Neuropeptide (calcitonin gene-related peptide) induction of nitric oxide in human keratinocytes in vitro. AB - Nitric oxide (NO) is an important signaling molecule in both the central nervous system and the periphery, where it is involved in neurotransmission, vascular and bronchial tone, inflammation, and cutaneous immune function. More recently, NO has been implicated in intracellular signaling and may have a role in cellular differentiation, cytokine expression, and apoptosis. The experiments described herein examined the effect of calcitonin gene-related protein (CGRP), a cutaneous nerve neuropeptide, on NO production in human keratinocytes in vitro. CGRP stimulated two distinct increases in NO production: one within 30 minutes and a second at 24 hours. CGRP stimulated a modest increase in inducible nitric oxide synthase (iNOS) at 3-6 hours. Experimental evidence suggested that CGRP stimulated both constitutive NOS activity and generation of NO via nitrosothiol degradation within the first hour. Production of NO was paralleled by a decrease in nitrosothiol levels for 2 hour, suggesting that immediate NO release may originate from pre-existing stores. Nitrosothiols are ubiquitous molecules that comprise an important NO pool and have intracellular regulatory roles, particularly linked to oxidative stress. The present data indicate that, in addition to its known cAMP signaling pathway, CGRP may act to regulate keratinocyte biology through intracellular NO by modulation of S-nitrosothiol stores and stimulation of NOS activity. PMID- 16710310 TI - Complete maternal isodisomy of chromosome 3 in a child with recessive dystrophic epidermolysis bullosa but no other phenotypic abnormalities. AB - The mechanobullous disease Hallopeau-Siemens recessive dystrophic epidermolysis bullosa (HS-RDEB) results from mutations in the type VII collagen gene (COL7A1) on chromosome 3p21.31. Typically, there are frameshift, splice site, or nonsense mutations on both alleles. In this report, we describe a patient with HS-RDEB, who was homozygous for a new frameshift mutation, 345insG, in exon 3 of COL7A1. However, sequencing of parental DNA showed that although the patient's mother was a heterozygous carrier of this mutation, the father's DNA contained only wild type sequence. Microsatellite marker analysis confirmed paternity and genotyping of 28 microsatellites spanning chromosome 3 revealed that the affected child was homozygous for every marker tested with all alleles originating from a single maternal chromosome 3. Thus, the HS-RDEB phenotype in this patient is due to complete maternal isodisomy of chromosome 3 and reduction to homozygosity of the mutant COL7A1 gene locus. To our knowledge, there are no published reports of uniparental disomy (UPD) in HS-RDEB; moreover, this case represents only the third example of UPD of chromosome 3 to be reported. The severity of the HS-RDEB in this case was similar to other affected individuals and no additional phenotypic abnormalities were observed, suggesting an absence of maternally imprinted genes on chromosome 3. PMID- 16710311 TI - Heteromeric nicotinic acetylcholine-dopamine autoreceptor complexes modulate striatal dopamine release. AB - In the striatum, dopamine and acetylcholine (ACh) modulate dopamine release by acting, respectively, on dopamine D(2) autoreceptors and nicotinic ACh (nACh) heteroreceptors localized on dopaminergic nerve terminals. The possibility that functional interactions exist between striatal D(2) autoreceptors and nACh receptors was studied with in vivo microdialysis in freely moving rats. Local perfusion of nicotine in the ventral striatum (shell of the nucleus accumbens) produced a marked increase in the extracellular levels of dopamine, which was completely counteracted by co-perfusion with either the non-alpha(7) nACh receptor antagonist dihydro-beta-erythroidine or the D(2-3) receptor agonist quinpirole. Local perfusion of the D(2-3) receptor antagonist raclopride produced an increase in the extracellular levels of dopamine, which was partially, but significantly, counteracted by coperfusion with dihydro-beta-erythroidine. These findings demonstrate a potent crosstalk between G protein-coupled receptors and ligand-gated ion channels in dopaminergic nerve terminals, with the D(2) autoreceptor modulating the efficacy of non-alpha(7) nACh receptor-mediated modulation of dopamine release. We further demonstrate physical interactions between beta(2) subunits of non-alpha(7) nicotinic acetylcholine receptors and D(2) autoreceptors in co-immunoprecipitation experiments with membrane preparations from co-transfected mammalian cells and rat striatum. These results reveal that striatal non-alpha(7) nicotinic acetylcholine receptors form part of heteromeric dopamine autoreceptor complexes that modulate dopamine release. PMID- 16710312 TI - GABA(B) receptor-positive modulation decreases selective molecular and behavioral effects of cocaine. AB - Exposure to cocaine induces selective behavioral and molecular adaptations. In rodents, acute cocaine induces increased locomotor activity, whereas prolonged drug exposure results in behavioral locomotor sensitization, which is thought to be a consequence of drug-induced neuroadaptive changes. Recent attention has been given to compounds activating GABA(B) receptors as potential antiaddictive therapies. In particular, the principle of allosteric positive GABA(B) receptor modulators is very promising in this respect, as positive modulators lack the sedative and muscle relaxant properties of full GABA(B) receptor agonists such as baclofen. Here, we investigated the effects of systemic application of the GABA(B) receptor-positive modulator GS39783 (N,N'-dicyclopentyl-2-methylsulfanyl 5-nitro-pyrimidine-4, 6-diamine) in animals treated with acute and chronic cocaine administration. Both GS39783 and baclofen dose dependently attenuated acute cocaine-induced hyperlocomotion. Furthermore, both compounds also efficiently blocked cocaine-induced Fos induction in the striatal complex. In chronic studies, GS39783 induced a modest attenuation of cocaine-induced locomotor sensitization. Chronic cocaine induces the accumulation of the transcription factor deltaFosB and upregulates cAMP-response-element-binding protein (CREB) and dopamine- and cAMP-regulated phosphoprotein of 32 kDa (DARPP 32). GS39783 blocked the induction/activation of DARPP-32 and CREB in the nucleus accumbens and dorsal striatum and partially inhibited deltaFosB accumulation in the dorsal striatum. In summary, our data provide evidence that GS39783 attenuates the acute behavioral effects of cocaine exposure in rodents and in addition prevents the induction of selective long-term adaptive changes in dopaminergic signaling pathways. Further investigation of GABA(B) receptor positive modulation as a novel therapeutic strategy for the treatment of cocaine dependence and possibly other drugs of abuse is therefore warranted. PMID- 16710314 TI - SLV313 (1-(2,3-dihydro-benzo[1,4]dioxin-5-yl)-4- [5-(4-fluoro-phenyl)-pyridin-3 ylmethyl]-piperazine monohydrochloride): a novel dopamine D2 receptor antagonist and 5-HT1A receptor agonist potential antipsychotic drug. AB - Combined dopamine D(2) receptor antagonism and serotonin (5-HT)(1A) receptor agonism may improve efficacy and alleviate some side effects associated with classical antipsychotics. The present study describes the in vitro and in vivo characterization of 1-(2,3-dihydro-benzo[1,4]dioxin-5-yl)-4-[5-(4-fluoro-phenyl) pyridin-3-ylmethyl]-piperazine monohydrochloride (SLV313), a D(2/3) antagonist and 5-HT(1A) agonist. SLV313 possessed high affinity at human recombinant D(2), D(3), D(4), 5-HT(2B), and 5-HT(1A) receptors, moderate affinity at 5-HT(7) and weak affinity at 5-HT(2A) receptors, with little-no affinity at 5-HT(4), 5-HT(6), alpha(1), and alpha(2) (rat), H(1) (guinea pig), M(1), M(4), 5-HT(3) receptors, and the 5-HT transporter. SLV313 had full agonist activity at cloned h5-HT(1A) receptors (pEC(50)=9.0) and full antagonist activity at hD(2) (pA(2)=9.3) and hD(3) (pA(2)=8.9) receptors. In vivo, SLV313 antagonized apomorphine-induced climbing and induced 5-HT(1A) syndrome behaviors and hypothermia, the latter behaviors being antagonized by the 5-HT(1A) antagonist WAY100635. In a drug discrimination procedure SLV313 induced full generalization to the training drug flesinoxan and was also antagonized by WAY100635. In the nucleus accumbens SLV313 reduced extracellular 5-HT and increased dopamine levels in the same dose range. Acetylcholine and dopamine were elevated in the hippocampus and mPFCx, the latter antagonized by WAY100635, suggesting possible 5-HT(1A)-dependent efficacy for the treatment of cognitive and attentional processes. SLV313 did not possess cataleptogenic potential (up to 60 mg/kg p.o.). The number of spontaneously active dopamine cells in the ventral tegmental area was reduced by SLV313 and clozapine, while no such changes were seen in the substantia nigra zona compacta following chronic administration. These results suggest that SLV313 is a full 5 HT(1A) receptor agonist and full D(2/3) receptor antagonist possessing characteristics of an atypical antipsychotic, representing a potential novel treatment for schizophrenia. PMID- 16710313 TI - Identifying genes in monoamine nuclei that may determine stress vulnerability and depressive behavior in Wistar-Kyoto rats. AB - The Wistar-Kyoto (WKY) rat is stress sensitive and exhibits depressive-like behavior. The locus coeruleus (LC)-norepinephrine and dorsal raphe (DR)-serotonin systems mediate certain aspects of the stress response and have been implicated in depression. Microarray technology was used to identify gene expression differences in the LC and DR between WKY vs Sprague-Dawley (SD) rats that might account for the WKY phenotype. RNA was isolated from microdissected LC and DR, amplified, and hybridized to microarrays (1 array/subject, n = 4/group). Significance of microarray (SAM) analysis revealed increased expression of 66 genes in the LC and 19 genes in the DR and decreased expression of 33 genes in the DR of WKY rats. Hierarchical clustering identified differences in gene expression profiles of WKY vs SD rats that generally concurred with SAM. Notably, genes that encoded for enzymes involved in norepinephrine turnover, amino-acid receptors, and certain G-protein-coupled receptors were elevated in the LC of WKY rats. The DR of WKY rats showed decreased expression of genes encoding several potassium channels and neurofilament genes. The chromosomal locations of 15 genes that were differentially expressed in WKY rats were near loci identified as contributing to depressive-like behaviors in the rat. The specific genes revealed by the present analysis as being differentially expressed in WKY rats may contribute to their unique behavioral profile and suggest targets that confer susceptibility to stress-related psychiatric disorders. PMID- 16710315 TI - Dopamine and benzodiazepine-dependent mechanisms regulate the EtOH-enhanced locomotor stimulation in the GABAA alpha1 subunit null mutant mice. AB - The present study investigated the role of the alpha1-containing GABA(A) receptors in the neurobehavioral actions of alcohol. In Experiment 1, mice lacking the alpha1 subunit (alpha1 (-/-)) were tested for their capacity to initiate operant-lever press responding for alcohol or sucrose. Alcohol intake in the home cage was also measured. In Experiment 2, the alpha1 (-/-) mice were injected with a range of alcohol doses (0.875-4.0 g/kg; i.p.) to evaluate the significance of the alpha1 subunit in alcohol's stimulant actions. In Experiment 3, we determined if the alcohol-induced stimulant effects were regulated via dopaminergic (DA) or benzodiazepine (BDZ)-dependent mechanisms. To accomplish this, we investigated the capacity of DA (eticlopride, SCH 23390) and BDZ (flumazenil, betaCCt) receptor antagonists to attenuate the alcohol-induced stimulant actions. Compared with wild-type mice (alpha1 (+/+)), the null mutants showed marked reductions in both EtOH and sucrose-maintained responding, and home cage alcohol drinking. The null mutants also showed significant increases in locomotor behaviors after injections of low-moderate alcohol doses (1.75-3.0 g/kg). betaCCt, flumazenil, eticlopride, and SCH 23390 were able to attenuate the alcohol-induced stimulation in mutant mice, in the absence of intrinsic effects. These data suggest the alpha1 receptor plays an important role in alcohol motivated behaviors; however, it also appears crucial in regulating the reinforcing properties associated with normal ingestive behaviors. Deleting the alpha1 subunit of the GABA(A) receptor appears to unmask alcohol's stimulatory effects; these effects appear to be regulated via an interaction of both DA- and GABA(A) BDZ-dependent mechanisms. PMID- 16710316 TI - Olanzapine-induced weight gain and increased visceral adiposity is blocked by melatonin replacement therapy in rats. AB - The atypical antipsychotic drug olanzapine increases body weight and visceral adiposity in schizophrenia. In rats, aging-associated increased body weight and visceral adiposity are reversed by administration of the pineal hormone melatonin. We asked if melatonin similarly would reverse olanzapine-induced increased weight and visceral adiposity in rats. Four groups (n=11/group) of female rats (240-250 g) were treated for 8 weeks with olanzapine, melatonin, olanzapine+melatonin, or vehicle alone in drinking water. Body weight and food and water consumption were determined weekly, locomotor activity at weeks 3 and 6, and nocturnal plasma melatonin concentration at week 7. At week 8, the rats were killed and visceral (perirenal, retroperitoneal, omental, and mesenteric) fat pads dissected and weighed. Olanzapine treatment reduced nocturnal plasma melatonin by 55% (p<0.001), which was restored to control levels by olanzapine+melatonin. Body weight increased 18% in rats treated with olanzapine alone, but only 10% with olanzapine+melatonin, 5% with melatonin alone, and 7% with vehicle control. Body weight and visceral fat pad weight increases in rats treated with olanzapine alone were greater than in each of the other three groups (all p<0.01), which were not significantly different. These results suggest that olanzapine-induced increases in body weight and visceral adiposity may be at least in part secondary to olanzapine-induced reduction of plasma melatonin levels, and that melatonin may be useful for the management of olanzapine-induced weight gain in humans. PMID- 16710317 TI - Involvement of the endocannabinoid system in the ability of long-term tricyclic antidepressant treatment to suppress stress-induced activation of the hypothalamic-pituitary-adrenal axis. AB - The efficacy of antidepressants has been linked in part to their ability to reduce activity of the hypothalamic-pituitary-adrenal (HPA) axis; however, the mechanism by which antidepressants regulate the HPA axis is largely unknown. Given that recent research has demonstrated that endocannabinoids can regulate the HPA axis and exhibit antidepressant potential, we examined the hypothesis that the endocannabinoid system is regulated by long-term antidepressant treatment. Three-week administration of the tricyclic antidepressant desipramine (10 mg/kg/day) resulted in a significant increase in the density of the cannabinoid CB(1) receptor in the hippocampus and hypothalamus, without significantly altering endocannabinoid content in any brain structure examined. Furthermore, chronic desipramine treatment resulted in a reduction in both secretion of corticosterone and the induction of the immediate early gene c-fos in the medial dorsal parvocellular region of the paraventricular nucleus of the hypothalamus (PVN) following a 5 min exposure to swim stress. Acute treatment with the CB(1) receptor antagonist, AM251 (1 mg/kg), before exposure to swim stress, completely occluded the ability of desipramine to reduce both corticosterone secretion and induction of c-fos expression in the PVN. Collectively, these data demonstrate that CB(1) receptor density in the hippocampus and hypothalamus is increased by chronic tricyclic antidepressant treatment, and suggest that this upregulation could contribute to the ability of tricyclic antidepressants to suppress stress-induced activation of the HPA axis. PMID- 16710318 TI - Sex- and histamine-dependent long-term cognitive effects of methamphetamine exposure. AB - As prenatal methamphetamine (MA) exposure results in long-term hippocampus dependent cognitive deficits, the increased MA use in women of childbearing age is of great concern. As mice are most commonly used in genetic models, we started to study the potential effects of neonatal MA exposure in female and male mice on brain function 3 months later. As histamine (HA) might mediate some effects of MA in adulthood, we also tested whether in neonates HA might mediate the long-term effects of MA using HA H(3) receptor agonists and antagonists. Stimulation of HA H(3) receptors by H(3) agonists inhibits HA synthesis and release, whereas inhibition of H(3) receptors by H(3) receptor antagonists increases HA release. MA (5 mg/kg), the H(3) receptor antagonist thioperamide (5 mg/kg), and the H(3) receptor agonist immepip (5 mg/kg) alone or in the presence of MA (5 mg/kg) were administered once daily from postnatal days 11 to 20 and the mice were tested at 3 months of age. Here we show that in mice exposure to MA early in life causes sex-dependent impairments in object recognition, spatial learning, and memory in the water maze, and pre-pulse inhibition in adulthood. HA mediates these impairments. Increasing HA release mimicked, whereas inhibiting HA release blocked the long-term detrimental MA effects. This model could be used to determine the role of genetic and environmental factors in MA-dependent cognitive impairments and to develop therapeutic strategies to inhibit them. PMID- 16710319 TI - Serotonin 2A -1438 G/A and G-protein Beta3 subunit C825T polymorphisms in patients with depression and SSRI-associated sexual side-effects. AB - The occurrence of sexual side-effects from antidepressants is thought to be mediated through serotonin 2A (5HT2A) receptors. It is currently unknown if functional polymorphisms in the 5HT2A receptor or its G-protein second messenger complex are related to sexual dysfunction in patients taking an selective serotonin reuptake inhibitor (SSRI) for depression. The purpose of this study was to determine the relationship of the 5HT2A -1438 G/A and GNB3 C825T single nucleotide polymorphisms with overall sexual well-being and individual components of sexual health as measured by the Changes in Sexual Functioning Questionnaire (CSFQ). We evaluated 89 outpatients (18-40 years of age) at low risk for other causes of sexual dysfunction who were being treated for depression with an SSRI and did not have sexual difficulties before taking the antidepressant. Outcome measures were stratified by 5HT2A and GNB3 genotypes. After controlling for age, gender, anxiety scale scores, and depression scale scores, persons with a GG genotype of the 5HT2A -1438 single nucleotide polymorphisms (SNP) were significantly more likely to be categorized as having sexual dysfunction than persons with a GA or AA genotype (OR=3.6; 95% CI 1.03, 12.6; p=0.046). Furthermore, the 5HT2A -1438 GG genotype was a significant predictor of lower arousal scores (p=0.022) after accounting for other measures. There was no significant relationship between any outcome measure and GNB3 genotype. PMID- 16710321 TI - Linking animal models of psychosis to computational models of dopamine function. AB - Psychosis is linked to dysregulation of the neuromodulator dopamine and antipsychotic drugs (APDs) work by blocking dopamine receptors. Dopamine modulated disruption of latent inhibition (LI) and conditioned avoidance response (CAR) have served as standard animal models of psychosis and antipsychotic action, respectively. Meanwhile, the 'temporal difference' algorithm (TD) has emerged as the leading computational model of dopamine neuron firing. In this report TD is extended to include action at the level of dopamine receptors in order to explain a number of behavioral phenomena including the dose-dependent disruption of CAR by APDs, the temporal dissociation of the effects of APDs on receptors vs behavior, the facilitation of LI by APDs, and the disruption of LI by amphetamine. The model also predicts an APD-induced change to the latency profile of CAR--a novel prediction that is verified experimentally. The model's primary contribution is to link dopamine neuron firing, receptor manipulation, and behavior within a common formal framework that may offer insights into clinical observations. PMID- 16710320 TI - Distinctive profiles of gene expression in the human nucleus accumbens associated with cocaine and heroin abuse. AB - Drug abuse is thought to induce long-term cellular and behavioral adaptations as a result of alterations in gene expression. Understanding the molecular consequences of addiction may contribute to the development of better treatment strategies. This study utilized high-throughput Affymetrix microarrays to identify gene expression changes in the post-mortem nucleus accumbens of chronic heroin abusers. These data were analyzed independently and in relation to our previously reported data involving human cocaine abusers, in order to determine which expression changes were drug specific and which may be common to the phenomenon of addiction. A significant decrease in the expression of numerous genes encoding proteins involved in presynaptic release of neurotransmitter was seen in heroin abusers, a finding not seen in the cocaine-abusing cohort. Conversely, the striking decrease in myelin-related genes observed in cocaine abusers was not evident in our cohort of heroin subjects. Overall, little overlap in gene expression profiles was seen between the two drug-abusing cohorts: out of the approximately 39,000 transcripts investigated, the abundance of only 25 was significantly changed in both cocaine and heroin abusers, with nearly one-half of these being altered in opposite directions. These data suggest that the profiles of nucleus accumbens gene expression associated with chronic heroin or cocaine abuse are largely unique, despite what are thought to be common effects of these drugs on dopamine neurotransmission in this brain region. A re-examination of our current assumptions about the commonality of molecular mechanisms associated with substance abuse seems warranted. PMID- 16710322 TI - Facilitation of cardiac vagal activity by CRF-R1 antagonists during swim stress in rats. AB - Exposure to stressors that elicit fear and feelings of hopelessness can cause severe vagal activation leading to bradycardia, syncope, and sudden death. These phenomena though documented, are difficult to diagnose, treat clinically, and prevent. Therefore, an animal model incorporating these cardiovascular conditions could be useful. The present study examined 'sinking' during a 2-h swim stress, a phenomenon that occurs in 50% of rats during 25 degrees C water exposure. Concurrent measurements of body temperature, immobility, heart rate (HR), and PR interval (a measure of vagal activity) were made. Neither decreases in immobility nor variations in hypothermia during swim were correlated with sinking. Bradycardia was more severe in sinking rats (average minimum HR+/-SEM; 143+/-13 vs 247+/-14; p<0.01), and PR interval was elevated (p<0.0001). To examine potential modulation of vagal activity during stress, corticotropin-relasing factor (CRF) receptor antagonists (antalarmin, R121919 and astressin B), a glucocorticoid receptor antagonist (RU486), and a peripherally acting cholinergic antagonist (methylatropine nitrate) were administered. The centrally acting CRF antagonist, antalarmin (32 mg/kg), produced elongation of the PR interval (p<0.0001), robust bradycardia (135+/-18; p<0.001), and increased sinking (92%; p<0.05), and methylatropine nitrate (3.2 mg/kg) blocked these effects. Corroborating these data, two different CRF antagonists, R121919 (30 mg/kg) and astressin B (intracerebroventricular (i.c.v.), 0.03 mug/rat) increased sinking to 100%. RU486 (20 mg/kg) blocked HPA axis negative feedback and decreased percent sinking to 25%. From these studies, we concluded that sinking during a 2-h water exposure was a result of extreme vagal hyperactivity. Furthermore, stress-induced CRF release may serve to protect against elevated cardiac vagal activity. PMID- 16710323 TI - Listeria monocytogenes: a multifaceted model. AB - The opportunistic intracellular pathogen Listeria monocytogenes has become a paradigm for the study of host-pathogen interactions and bacterial adaptation to mammalian hosts. Analysis of L. monocytogenes infection has provided considerable insight into how bacteria invade cells, move intracellularly, and disseminate in tissues, as well as tools to address fundamental processes in cell biology. Moreover, the vast amount of knowledge that has been gathered through in-depth comparative genomic analyses and in vivo studies makes L. monocytogenes one of the most well-studied bacterial pathogens. PMID- 16710324 TI - Signalling and oxidant adaptation in Candida albicans and Aspergillus fumigatus. AB - Candida species and Aspergillus fumigatus were once thought to be relatively benign organisms. However, it is now known that this is not the case - Candida species rank among the top four causes of nosocomial infectious diseases in humans and A. fumigatus is the most deadly mould, often having a 90% mortality rate in immunocompromised transplant recipients. Adaptation to stress, including oxidative stress, is a necessary requisite for survival of these organisms during infection. Here, we describe the latest information on the signalling pathways and target proteins that contribute to oxidant adaptation in C. albicans and A. fumigatus, which has been obtained primarily through the analysis of mutants or inference from genome annotation. PMID- 16710325 TI - The interaction of bacterial pathogens with platelets. AB - In recent years, the frequency of serious cardiovascular infections such as endocarditis has increased, particularly in association with nosocomially acquired antibiotic-resistant pathogens. Growing evidence suggests a crucial role for the interaction of bacteria with human platelets in the pathogenesis of cardiovascular infections. Here, we review the nature of the interactions between platelets and bacteria, and the role of these interactions in the pathogenesis of endocarditis and other cardiovascular diseases. PMID- 16710326 TI - Advances in tuberculosis vaccine strategies. AB - Tuberculosis (TB), an ancient human scourge, is a growing health problem in the developing world. Approximately two million deaths each year are caused by TB, which is the leading cause of death in HIV-infected individuals. Clearly, an improved TB vaccine is desperately needed. Heterologous prime-boost regimens probably represent the best hope for an improved vaccine regimen to prevent TB. This first generation of new vaccines might also complement drug treatment regimens and be effective against reactivation of TB from the latent state, which would significantly enhance their usefulness. PMID- 16710327 TI - The trypanolytic factor of human serum. AB - African trypanosomes (the prototype of which is Trypanosoma brucei brucei) are protozoan parasites that infect a wide range of mammals. Human blood, unlike the blood of other mammals, has efficient trypanolytic activity, and this needs to be counteracted by these parasites. Resistance to this activity has arisen in two subspecies of Trypanosoma brucei - Trypanosoma brucei rhodesiense and Trypanosoma brucei gambiense - allowing these parasites to infect humans, and this results in sleeping sickness in East Africa and West Africa, respectively. Study of the mechanism by which T. b. rhodesiense escapes lysis by human serum led to the identification of an ionic-pore-forming apolipoprotein - known as apolipoprotein L1 - that is associated with high-density-lipoprotein particles in human blood. In this Opinion article, we argue that apolipoprotein L1 is the factor that is responsible for the trypanolytic activity of human serum. PMID- 16710329 TI - Our paper anniversary. PMID- 16710330 TI - Making carbon-nitrogen bonds in biological and chemical synthesis. AB - The function of many biologically active molecules requires the presence of carbon-nitrogen bonds in strategic positions. The biosynthetic pathways leading to such bonds can be bypassed through chemical synthesis to synthesize natural products more efficiently and also to generate the molecular diversity unavailable in nature. PMID- 16710331 TI - A century at the chemistry-biology interface. AB - The field of chemical biology is now hitting its stride. Chemical biologists have developed essential tools that are being used to illuminate complex cellular events. The application of chemical principles to biological phenomena has revealed new opportunities for drug discovery. This report highlights recent progress and exciting new directions in chemical genetics and drug discovery. PMID- 16710332 TI - Jeremy Knowles. PMID- 16710333 TI - Snapshots of three intermediates at the active site of pyruvate oxidase. PMID- 16710334 TI - Signalomic signatures enlighten drug profiling. PMID- 16710335 TI - Nucleation of huntingtin aggregation in cells. PMID- 16710336 TI - Wanting contact: how to pick up a channel. PMID- 16710337 TI - Chemical knockout of C-reactive protein in cardiovascular disease. PMID- 16710339 TI - Modular approaches to expanding the functions of living matter. AB - The synthesis of increasingly complex unnatural networks embedded in living matter is an emerging theme in synthetic biology. Synthetic networks have allowed the creation of organisms endowed with toggle switches, logic gates, pattern forming systems, oscillators, cellular sensors, new modes of gene regulation and expanded genetic codes. A common challenge of this work is the addition of specific new functions to complex living organisms. This requires spatial and temporal control of molecular interactions and fluxes to achieve the desired outcomes. Here we review recent successes in this emerging field and discuss strategies for addressing the challenges of increasing network complexity. PMID- 16710344 TI - Oncolytic adenoviral vector carrying the cytosine deaminase gene for melanoma gene therapy. AB - We constructed an oncolytic adenoviral vector Ad.HE1HCD3, in which the adenoviral E1A promoter was replaced by a human tyrosinase enhancer (HTE)/promoter. The RGD 4C peptide was inserted into the HI loop of the fiber knob domain to increase the transduction efficiency of this vector for tumor cell lines. We also inserted the prodrug activating cytosine deaminase gene driven by the HTE/promoter into the E3 region of the Ad.HE1HCD3 vector. The in vitro cytotoxic effect of the Ad.HE1HCD3 vector with 5-fluorocytosine (5-FC) was greater than that of a wild-type adenovirus or that of the Ad.HE1HCD3 vector alone in tyrosinase-positive melanoma cell lines at low multiplicity of infection. Intratumoral injection of low doses of the Ad.HE1HCD3 vector into xenotransplanted human melanoma cell lines followed by the intraperitoneal injection of 5-FC led to a greater degree of tumor regression in vivo than did the intratumoral injection of the same dose of the Ad.HE1HCD3 vector alone. This oncolytic vector with a melanoma-specific prodrug activation therapeutic transcription unit and a RGD targeted fiber protein offers a potent therapeutic combination for the gene therapy of melanoma. PMID- 16710346 TI - Prevention of radiation-induced pneumonitis by recombinant adenovirus-mediated transferring of soluble TGF-beta type II receptor gene. AB - To investigate whether radiation-induced pneumonitis in the mouse-irradiated lung could be prevented by recombinant adenovirus-mediated soluble transforming growth factor-beta (TGF-beta) type II receptor gene therapy. Radiation fibrosis-prone mice (C57BL/6J) were randomly divided into four groups consisting of a (1) control group (sham-irradiated); (2) radiation (RT)-alone group; (3) RT+AdCMVsTbetaR group and (4) RT+AdCMVluc group. The RT-alone and sham-irradiated mice were killed at several time points after thoracic irradiation with a single dose of 9 Gy, and then the TGF-beta1 concentrations in serum and broncho-alveolar lavage fluid (BALF) were quantified by enzyme-linked immunosorbent assay (ELISA). We used an adenoviral vector expressing a soluble TGF-beta type II receptor (AdCMVsTbetaR), which can bind to TGF-beta and then block the TGF-beta receptor mediated signal transduction. The C57BL/6J mice were intraperitoneally (i.p.) injected with either 5 x 10(8) plaque-forming units of AdCMVsTbetaR or AdCMVluc, a control adenovirus-expressing luciferase, a week preceding and a week following the X-ray thoracic irradiation. Four weeks after irradiation, the mice were killed and the concentration of TGF-beta1 in the serum and BALF were then measured using ELISA and the lung tissue specimens were examined histopathologically. Following thoracic irradiation with a single dose of 9 Gy, radiation-induced TGF-beta1 release in the serum reached the first peak concentration at 12 h and then declined. It reached a maximal value at 2 weeks after irradiation. In the BALF, the TGF-beta1 concentration was appreciable within the first hour and thereafter declined. It reached a maximal value at 3 days after irradiation. A one-time i.p. injection of AdCMVsTbetaR 1 week before irradiation could not completely suppress the two peaks of the radiation-induced TGF-beta1 increase, whereas an injection a week preceding and a week following thoracic irradiation was able to suppress those two peaks thoroughly. The TGF beta1 was completely suppressed in the AdCMVsTbetaR-treated mouse serum and BALF; however, no statistical difference was observed in the serum and BALF between the AdCMVluc-infected mice and the control mice at 4 weeks after irradiation (P < 0.05). A histopathological examination showed only mild radiation pneumonitis in the irradiated lungs of AdCMVsTbetaR-treated mice in comparison to the AdCMVluc infected and RT-alone mice. Our results demonstrated that TGF-beta1 plays an important role in radiation pneumonitis, thus suggesting that the adenovirus mediated overexpression in soluble TGF-beta type II receptor gene therapy may be a potentially feasible and effective strategy for the prevention of radiation pneumonitis. PMID- 16710345 TI - Phase I/II trial of melanoma therapy with dendritic cells transfected with autologous tumor-mRNA. AB - We have developed an individualized melanoma vaccine based on transfection of autologous dendritic cells (DCs) with autologous tumor-mRNA. Dendritic cells loaded with complete tumor-mRNA may generate an immune response against a broad repertoire of antigens, including unique patient-specific antigens. The purpose of the present phase I/II trial was to evaluate the feasibility and safety of the vaccine, and the ability of the DCs to elicit T-cell responses in melanoma patients. Further, we compared intradermal (i.d.) and intranodal (i.n.) vaccine administration. Twenty-two patients with advanced malignant melanoma were included, each receiving four weekly vaccines. Monocyte-derived DCs were transfected with tumor-mRNA by electroporation, matured and cryopreserved. We obtained successful vaccine production for all patients elected. No serious adverse effects were observed. A vaccine-specific immune response was demonstrated in 9/19 patients evaluable by T-cell assays (T-cell proliferation/interferon-gamma ELISPOT) and in 8/18 patients evaluable by delayed type hypersensitivity (DTH) reaction. The response was demonstrated in 7/10 patients vaccinated intradermally and in 3/12 patients vaccinated intranodally. We conclude that immuno-gene-therapy with the described DC-vaccine is feasible and safe, and that the vaccine can elicit in vivo T-cell responses against antigens encoded by the transfected tumor-mRNA. The response rates do not suggest an advantage in applying i.n. vaccination. PMID- 16710347 TI - Transcriptional targeting of adenovirus vectors with the squamous cell carcinoma specific antigen-2 promoter for selective apoptosis induction in lung cancer. AB - Squamous cell carcinoma antigens SCCA1 and SCCA2 are highly homologous serine proteinase inhibitors which have been widely utilized as serological markers for squamous cell cancers, but it has recently been demonstrated that only SCCA2 is truly specific for certain forms of lung cancer. Using a construct containing the 5'-flanking region of the SCCA2 gene between -460 and +0 bp and the luciferase reporter gene, SCCA2 promoter activity was detected in SCCA2-producing SCC cell lines (LK-2, LC-1), but not in SCCA2-nonproducing lung adenocarcinoma cell lines (A549, ABC-1, and RERF-LC-MS) or normal cells (WI-38, SAEC, and NHEK-Adult). Infection with a recombinant adenovirus vector, Ad-SCCA2-DsRed, resulted in cell specific expression of the SCCA2 promoter-driven DsRed marker gene only in LK-2 and LC-1 cells. The same strategy was used for SCCA2-driven expression of a proapoptotic gene, (KLAKLAK)2, which can cause mitochondrial disruption by triggering mitochondrial permeabilization and swelling, resulting in the release of cytochrome c and induction of apoptosis. Infection with Ad-SCCA2-KLAKLAK2 specifically reduced the growth of the two human lung SCC cell lines compared to the SCCA2 nonproducing cell lines both in vitro and in vivo, suggesting that the SCCA2 promoter had a tumor-specific effect. These results suggest that transduction of SCCA2 promoter-controlled suicide genes by adenoviral vectors can confer transcriptionally targeted cytotoxicity in SCCA2-producing lung SCC cells, and represents a novel strategy for gene transfer specifically targeted to SCC in the lung. PMID- 16710348 TI - Urinary IL-18 is an early predictive biomarker of acute kidney injury after cardiac surgery. AB - Acute kidney injury (AKI) is a frequent complication of cardiopulmonary bypass (CPB). The lack of early biomarkers for AKI has impaired our ability to intervene in a timely manner. Urinary neutrophil gelatinase-associated lipocalin (NGAL) is recently demonstrated as an early biomarker of AKI after CPB, increasing 25-fold within 2 h and declining 6 h after surgery. In the present study, we tested whether interleukin-18 (IL-18) is a predictive biomarker for AKI in the same group of patients following CPB. Exclusion criteria included pre-existing renal insufficiency and nephrotoxin use. Serial urine samples were analyzed by enzyme linked immunosorbent assay for IL-18 in 20 patients who developed AKI (defined as a 50% or greater increase in serum creatinine after CPB) and 35 controls (age, race, and gender-matched patients who did not develop AKI after CPB). Using serum creatinine, AKI was detected only 48-72 h after CPB. In contrast, urine IL-18 increased at 4-6 h after CPB, peaked at over 25-fold at 12 h, and remained markedly elevated up to 48 h after CPB. The performance of IL-18 as demonstrated by area under the receiver operating characteristics curve for diagnosis of AKI at 4, 12, and 24 h after CPB was 61, 75, and 73% respectively. Also, on multivariate analysis, both IL-18 and NGAL were independently associated with number of days in AKI among cases. Our results indicate that IL-18 is an early, predictive biomarker of AKI after CPB, and that NGAL and IL-18 are increased in tandem after CPB. The combination of these two biomarkers may allow for the reliable early diagnosis and prognosis of AKI at all times after CPB, much before the rise in serum creatinine. PMID- 16710349 TI - Simvastatin maintains steady patterns of GFR and improves AER and expression of slit diaphragm proteins in type II diabetes. AB - The factors determining the course of glomerular filtration rate (GFR) and albumin excretion rate (AER) and the expression of mRNA of slit diaphragm (SD) and podocyte proteins in microalbuminuric, hypertensive type II diabetic patients are not fully understood. GFR, AER, and SD protein mRNA were studied in 86 microalbuminuric, hypertensive, type II diabetics at baseline and after 4-year random double-blind treatment either with 40 mg simvastatin (Group 1) or with 30 g cholestyramine (Group 2) per day. Both groups had at baseline a GFR decay per year in the previous 2-4 years of 3 ml/min/1.73 m(2). Both Groups 1 and 2 showed a significant decrease of low-density lipoprotein cholesterol levels after simvastatin and cholestyramine treatment (P<0.01). No change from base line values was observed as for hs-C-reactive protein and interleukin-6. A significant decrease of 8-hydroxydeoxyguanosine urinary excretion was observed after simvastatin treatment. GFR did not change from baseline with simvstatin, whereas a decrease was observed with cholestyramine treatment (simvastatin vs cholestyramine: -0.21 vs -2.75 ml/min/1.73 m(2), P<0.01). AER decreased in Group 1 (P<0.01), but not in Group 2 patients. Real-time polymerase chain reaction measurement of mRNA SD proteins (CD2AP, FAT, Actn 4, NPHS1, and NPHS2) significantly increased in kidney biopsy specimens after simvastatin, but not cholestyramine treatment. Simvastatin, but not cholestyramine, 4-year treatment maintains steady patterns of GFR, and improves AER and expression of SD proteins in type II diabetes, despite similar hypocholesterolemic effects in circulation. PMID- 16710350 TI - Effect of spironolactone and captopril on nitric oxide and S-nitrosothiol formation in kidney of L-NAME-treated rats. AB - Although angiotensin-converting enzyme (ACE) inhibitors are well-established drugs in the treatment of hypertension, they are not supposed to be sufficient in the inhibition of aldosterone formation. The present study analyzes the effect of aldosterone receptor antagonist, spironolactone and ACE inhibitor, captopril on nitric oxide (NO) and S-nitrosothiol formation in the kidney of N(G)-nitro-L arginine methyl ester (L-NAME)-treated rats. Male Wistar rats were divided into six groups: (1) controls, (2) L-NAME (40 mg/kg/day), (3) spironolactone (200 mg/kg/day), (4) captopril (100 mg/kg/day), (5) L-NAME+spironolactone, and (6) L NAME+captopril. After 4 weeks, NO synthase (NOS) activity, protein expression of endothelial NOS, inducible NOS and concentration of thiol and S-nitrosothiol groups were determined in the kidney. Besides the increase in systolic blood pressure (by 32%) and the decrease in NOS activity (by 37%), L-NAME treatment lowered the concentration of thiols (by 32%) and S-nitrosothiols (by 36%) in the renal tissue. Simultaneous treatment with spironolactone preserved NOS activity and S-nitrosothiols on the control level, whereas captopril did not affect these parameters modified by L-NAME treatment. Moreover, spironolactone increased expression of endothelial NOS protein without affecting inducible NOS protein expression. In conclusion, both captopril and spironolactone prevented L-NAME induced hypertension and the decline of the antioxidant potential of the kidney tissue. However, only spironolactone improved NOS activity which led to the S nitrosothiols formation. Both NO itself and S-nitrosothiols may contribute to the preventive effect of spironolactone against development of L-NAME-induced hypertension. PMID- 16710351 TI - Laser Doppler flowmetry detection of endothelial dysfunction in end-stage renal disease patients: correlation with cardiovascular risk. AB - Prediction of cardiovascular (CV) complications represents the Achilles' heel of end-stage renal disease. Surrogate markers of endothelial dysfunction have been advocated as predictors of CV risk in this cohort of patients. We have recently adapted a noninvasive laser Doppler flowmetry (LDF) functional testing of endothelium-dependent microvascular reactivity and demonstrated that end-stage renal disease patients are characterized by profound alterations in thermal hyperemic responsiveness. We hypothesized that such functional assessment of the cutaneous microcirculation may offer a valid, noninvasive test of the severity of endothelial dysfunction and CV risk. To test this hypothesis, we performed a cross-sectional study, in which we compared LDF measurements to conventional risk factors, and performed a pilot longitudinal study. LDF studies were performed in 70 patients and 33 controls. Framingham and Cardiorisk scores were near equivalent for low-risk patients, but more divergent as risk increased. C reactive protein (CRP) levels and LDF parameters (amplitude of thermal hyperemia (TH), area under the curve of TH) showed significant abnormality in high-risk vs low-risk patients calculated using either Framingham or Cardiorisk scores. Patients who had abnormal LDF parameters showed increased CV mortality, however, had similar risk assessments (Framingham, Cardiorisk, CRP, and homocysteine) to those with unimpaired LDF tracings. In conclusion, LDF parameters of microvascular reactivity offer a sensitive characterization of endothelial dysfunction, which may improve CV risk assessment through incorporation into the Framingham or Cardiorisk algorithm. PMID- 16710352 TI - HGF gene therapy attenuates renal allograft scarring by preventing the profibrotic inflammatory-induced mechanisms. AB - Inflammatory processes and tissue scarring are characteristic features of chronic allograft nephropathy. Hepatocyte growth factor (HGF) has beneficial effects on renal fibrosis and it also ameliorates renal interstitial inflammation as it has been recently described. Contrarily to protein administration, intramuscular gene electrotransfer allows sustained release of HGF. So, here we hypothesized that gene therapy with human HGF would diminish the characteristic scarring of chronic allograft nephropathy either by antagonizing tissue fibrosis mechanisms or by reducing inflammation. Lewis rats transplanted with cold preserved Fischer kidneys received vehicle (NoHGF) or intramuscular plasmid DNA encoding HGF plus electroporation either before transplantation (IniHGF, early post-transplant cytoprotection of tubular cells) or 8/10 weeks after transplantation (DelHGF, delayed prevention of chronic mechanisms). Serum creatinine and proteinuria were measured every 4 weeks for 24 weeks. Grafts at 12 or 24 weeks were evaluated for glomerulosclerosis, fibrosis inflammatory cells and mediators, cell regeneration and tubulo-interstitial damage. Nontreated animals developed renal insufficiency, progressive proteinuria and fibrosis among other characteristic histological features of chronic allograft nephropathy. Treatment with human HGF, especially when delayed until the onset of fibrogenic mechanisms, reduced renal failure and mortality, diminished tubule-interstitial damage, induced cell regeneration, decreased inflammation, NF-kappaB activation, and profibrotic markers at 12 weeks and prevented late interstitial fibrosis and glomerulosclerosis. The effectiveness of HGF-gene therapy in the prevention of renal allograft scarring is related with the halt of profibrotic inflammatory-induced mechanisms. PMID- 16710353 TI - Developmental expression of ACE2 in the SHR kidney: a role in hypertension? AB - The abnormal development of the intrarenal renin-angiotensin system (RAS) is thought contribute to adult-onset hypertension in the spontaneously hypertensive rat (SHR). Angiotensin-converting enzyme 2 (ACE2) is a novel enzyme with complementary actions to that of ACE. Recent studies have shown that ACE2 expression is reduced in the adult SHR. However, its regulation in pre hypertensive animals is unknown. In this study, we examine the developmental expression of ACE2 in the rodent kidney and its temporal expression, as it relates to the development of hypertension in the SHR model. Kidneys from SHR and normotensive Wistar Kyoto (WKY) rats (n=8-12/group) at birth, 6 weeks of age, and adulthood (80 days) were examined. Gene expression and activity of ACE2 were determined by real-time reverse transcription-polymerase chain reaction and quenched fluorescence assays, respectively. Renal expression was localized by in situ hybridization and immunohistochemistry. The expression and ACE2 activity are significantly increased in the SHR kidney at birth. With the onset of hypertension, the tubular expression of ACE2 falls in SHR compared to WKY and remains reduced in the adult SHR kidney. Glomerular expression is paradoxically increased in the SHR glomerulus. The overall developmental pattern of ACE2 expression in the SHR kidney is also modified, with declining expression over the course of renal development. The developmental pattern of ACE2 expression in the SHR kidney is altered before the onset of hypertension, consistent with the key role of the RAS in the pathogenesis of adult-onset hypertension. Further research is required to distinguish the contribution of these changes to the development and progression of hypertension in this model. PMID- 16710354 TI - Hypoxia-inducible factors and tubular cell survival in isolated perfused kidneys. AB - Adaptation to hypoxic environment is conferred through hypoxia-inducible transcription factors (HIFs). We have previously shown that the HIF system is transiently activated in vivo in radiocontrast-induced acute renal failure, associated with profound hypoxia in the renal medulla. Medullary thick ascending limbs (mTALs), the most affected nephron segments in this model, were virtually unable to mount an adaptive HIF response. Here, we study correlations between oxygenation, HIF activation, and cell viability in a related ex vivo model, the isolated perfused rat kidney (IPK). In IPKs perfused with cell-free oxygenated medium, severe medullary hypoxic damage developed, affecting 42+/-9% of mTALs in the mid-inner stripe. HIF-1alpha tubular immunostaining was noted with a zonal and tubular pattern largely similar to our findings in vivo: in 34+/-3% of collecting ducts (CDs) within the mid-inner stripe and extensively in the papillary tip, whereas mTALs were all HIF-negative. In IPKs supplemented with RBCs (improved oxygen supply), mTAL damage was totally prevented and CDs' HIF expression was attenuated (22+/-4%). By contrast, although measures designed to reduce medullary hypoxia by decreasing tubular reabsorptive activity (furosemide, ouabain, or high-albumin-non-filtering system) reduced mTAL damage, all paradoxically resulted in increased HIF expression in CDs (51+/-4%), and 17+/-3% of mTALs became immunostained as well. Our data confirm that CDs and mTALs have markedly different HIF responses, which correlate with their viability under hypoxic stress. mTALs transcriptional adaptation occurs within a narrow hypoxic range, and it appears that workload reduction can shift mTALs into this window of opportunity for HIF activation and survival. PMID- 16710355 TI - Maxi-K channels contribute to urinary potassium excretion in the ROMK-deficient mouse model of Type II Bartter's syndrome and in adaptation to a high-K diet. AB - Type II Bartter's syndrome is a hereditary hypokalemic renal salt-wasting disorder caused by mutations in the ROMK channel (Kir1.1; Kcnj1), mediating potassium recycling in the thick ascending limb of Henle's loop (TAL) and potassium secretion in the distal tubule and cortical collecting duct (CCT). Newborns with Type II Bartter are transiently hyperkalemic, consistent with loss of ROMK channel function in potassium secretion in distal convoluted tubule and CCT. Yet, these infants rapidly develop persistent hypokalemia owing to increased renal potassium excretion mediated by unknown mechanisms. Here, we used free-flow micropuncture and stationary microperfusion of the late distal tubule to explore the mechanism of renal potassium wasting in the Romk-deficient, Type II Bartter's mouse. We show that potassium absorption in the loop of Henle is reduced in Romk deficient mice and can account for a significant fraction of renal potassium loss. In addition, we show that iberiotoxin (IBTX)-sensitive, flow-stimulated maxi-K channels account for sustained potassium secretion in the late distal tubule, despite loss of ROMK function. IBTX-sensitive potassium secretion is also increased in high-potassium-adapted wild-type mice. Thus, renal potassium wasting in Type II Bartter is due to both reduced reabsorption in the TAL and K secretion by max-K channels in the late distal tubule. PMID- 16710356 TI - Comparison between valsartan and valsartan plus cilnidipine in type II diabetics with normo- and microalbuminuria. AB - Cilnidipine, an L-/N-type calcium channel blocker, dilates the efferent glomerular arterioles in an experimental model and shows a renoprotective effect, but its clinical benefits and safety have not yet been assessed in type II diabetics with albuminuria. The objective of this trial was to evaluate the effect of reducing albuminuria in type II diabetic patients with a combination therapy consisting of valsartan plus cilnidipine versus monotherapy with valsartan. An open-label, randomized controlled trial was conducted from April 2002 to October 2003 in 87 Japanese patients aged 31-90 years with type II diabetes showing albuminuria (urinary albumin/creatinine ratio: 10-300 mg/g). The patients were randomized to receive either valsartan (n=41) or valsartan plus cilnidipine (n=46) once daily for 1 year. The primary end point was the percent change in the albumin/creatinine ratio. The secondary end points were the progression/regression of albuminuria, blood pressure (BP), renal function, and safety. After 1 year, the albumin/creatinine ratio was found to have decreased more markedly in the valsartan plus cilnidipine group than in the valsartan group (reduction rate -44+/-11% (s.e.) versus -9+/-7% (s.e.); P=0.014 by analysis of covariance). Although a significant reduction was observed in the systolic and diastolic BP of both groups from baseline to 1 year (P<0.0001, respectively), there was no significant difference in the change in the BP between the two groups (systolic BP, P=0.066; diastolic BP, P=0.391). There were also no significant differences in the side effects between the two groups. Cilnidipine was thus found to show an additive effect with valsartan and thereby caused a reduction in albuminuria in type II diabetics. PMID- 16710357 TI - Renal crystal deposits and histopathology in patients with cystine stones. AB - We have biopsied the papillae of patients who have cystine stones asking if this stone type is associated with specific tissue changes. We studied seven cystine stone formers (SF) treated with percutaneous nephrolithotomy using digital video imaging of renal papillae for mapping and obtained papillary biopsies. Biopsies were analyzed by routine light and electron microscopy, infrared spectroscopy, electron diffraction, and micro-CT. Many ducts of Bellini (BD) had an enlarged ostium, and all such were plugged with cystine crystals, and had injured or absent lining cells with a surrounding interstitium that was inflamed to fibrotic. Crystal plugs often projected into the urinary space. Many inner medullary collecting ducts (IMCD) were dilated with or without crystal plugging. Apatite crystals were identified in the lumens of loops of Henle and IMCD. Abundance of interstitial Randall's plaque was equivalent in amount to that of non-SF. In the cortex, glomerular obsolescence and interstitial fibrosis exceeded normal. Cystine crystallizes in BD with the probable result of cell injury, interstitial reaction, nephron obstruction, and with the potential of inducing cortical change and loss of IMCD tubular fluid pH regulation, resulting in apatite formation. The pattern of IMCD dilation, and loss of medullary structures is most compatible with such obstruction, either from BD lumen plugs or urinary tract obstruction from stones themselves. PMID- 16710358 TI - Unraveling MCL-1 degradation. PMID- 16710359 TI - A novel cellular survival factor--the B2 subunit of vacuolar H+-ATPase inhibits apoptosis. AB - The ubiquitous vacuolar H(+)-ATPase, a multisubunit proton pump, is essential for intraorganellar acidification. Disruption of its function leads to disturbances of organelle function and cell death. Here, we report that overexpression of the B2 subunit of the H(+)-ATPase inhibits apoptosis. This antiapoptotic effect is not mediated by an increase in H(+)-ATPase activity but through activation of the Ras-mitogen-activated protein kinase (MAPK)-signaling pathway that results in the serine phosphorylation of Bad at residues 112 and 155. Increased Bad phosphorylation reduces its translocation to mitochondria, limits the release of mitochondrial cytochrome c and apoptosis-inducing factor and increases the resistance of the B2 overexpressing cells to apoptosis. Screening experiments of kinase inhibitors, including inhibitors of cAMP-activated protein kinase, protein kinase C, protein kinase B, (MAPK/extracellular signal-regulated (ERK) kinase) MEK and Ste-MEK1(13), a cell permeable ERK activation inhibitor peptide, revealed that the B2 subunit of H(+)-ATPase acts upstream of MEK activation in the MEK/ERK pathway to ameliorate apoptosis. PMID- 16710360 TI - Activation of extracellular signal-regulated protein kinase 5 downregulates FasL upon osmotic stress. AB - Extracellular signal-regulated protein kinase (ERK) 5 is a mitogen-activated protein kinase (MAPK) that is activated by dual phosphorylation via a unique MAPK/ERK kinase 5, MEK5. The physiological importance of this signaling cascade is underscored by the early embryonic death caused by the targeted deletion of the erk5 or the mek5 genes in mice. Here, we have found that ERK5 is required for mediating the survival of fibroblasts under basal conditions and in response to sorbitol treatment. Increased Fas ligand (FasL) expression acts as a positive feedback loop to enhance apoptosis of ERK5- or MEK5-deficient cells under conditions of osmotic stress. Compared to wild-type cells, erk5-/- and mek5-/- fibroblasts treated with sorbitol display a reduced protein kinase B (PKB) activity associated with increased Forkhead box O3a (Foxo3a) activity. Based on these results, we conclude that the ERK5 signaling pathway promotes cell survival by downregulating FasL expression via a mechanism that implicates PKB-dependent inhibition of Foxo3a downstream of phosphoinositide 3 kinase. PMID- 16710361 TI - FADD self-association is required for stable interaction with an activated death receptor. AB - Receptor-mediated programmed cell death proceeds through an activated receptor to which the death adaptor FADD and the initiator procaspases 8 and/or 10 are recruited following receptor stimulation. The adaptor FADD is responsible for both receptor binding and recruitment of the procaspases into the death-inducing signaling complex. Biochemical dissection of the FADD death effector domain and functional replacement with a coiled-coil motif demonstrates that there is an obligatory FADD self-association via the DED during assembly of the death inducing signaling complex. Using engineered oligomerization motifs with defined stoichiometries, the requirement for FADD self-association through the DED can be separated from the caspase-recruitment function of the domain. Disruption of FADD self-association precludes formation of a competent signaling complex. On this basis, we propose an alternative architecture for the FADD signaling complex in which FADD acts as a molecular bridge to stitch together an array of activated death receptors. PMID- 16710362 TI - Mitochondrial outer membrane permeabilization during apoptosis: the innocent bystander scenario. AB - Mitochondrial outer membrane permeabilization (MOMP) is considered the 'point of no return' as this event is responsible for engaging the apoptotic cascade in numerous cell death pathways. MOMP is directly governed by a subset of the BCL-2 family of proapoptotic proteins, which induce disruptions in the outer mitochondrial membrane (OMM) and subsequent release of death-promoting proteins like cytochrome c. The proposal here is centered on our hypothesis that MOMP is dictated by an interaction between the cytosol and the OMM, and although proteins of the OMM may be important in the process, the 'decision' to undergo apoptosis originates within the cytosol with no participation (in terms of yes, no and when) by mitochondria. PMID- 16710364 TI - Regulated targeting of Bax and Bak to intracellular membranes during apoptosis. PMID- 16710363 TI - The p53-Bcl-2 connection. PMID- 16710365 TI - Multifunctional reaper: sixty-five amino acids of fury. PMID- 16710366 TI - Identification of the death zone: a spatially restricted region for programmed cell death that sculpts the fly eye. AB - Programmed cell death (PCD) sculpts many developing tissues. The final patterning step of the Drosophila retina is the elimination, through PCD, of a subset of interommatidial lattice cells during pupation. It is not understood how this process is spatially regulated to ensure that cells die in the proper positions. To address this, we observed PCD of lattice cells in the pupal retina in real time. This live-visualization method demonstrates that lattice cell apoptosis is a highly specific process. In all, 85% of lattice cells die in exclusive 'death zone' positions between adjacent ommatidia. In contrast, cells that make specific contacts with primary pigment cells are protected from death. Two signaling pathways, Drosophila epidermal growth factor receptor (dEgfr) and Notch, that are thought to be central to the regulation of lattice cell survival and death, are not sufficient to establish the death zone. Thus, application of live visualization to the fly eye gives new insight into a dynamic developmental process. PMID- 16710367 TI - Bcl2a1 serves as a switch in death of motor neurons in amyotrophic lateral sclerosis. PMID- 16710373 TI - Infection biology. PMID- 16710369 TI - Immunology: Toll-like receptors and antibody responses. AB - Microbial components, such as lipopolysaccharides, augment immune responses by activating Toll-like receptors (TLRs). Some have interpreted this to mean that TLR signalling might not only help to initiate the adaptive immune response, but may also be required for it. The expanded view is shared by Pasare and Medzhitov, who conclude from an analysis of mice deficient in MyD88 (a TLR-signalling adaptor protein) that the generation of T-dependent antigen-specific antibody responses requires activation of TLRs in B cells. However, we show here that robust antibody responses can be elicited even in the absence of TLR signals. This appreciable TLR-independence of immune responses should be taken into account in the rational design of immunogenic and toleragenic vaccines. PMID- 16710374 TI - No easy answers. PMID- 16710375 TI - Patents for the people. PMID- 16710377 TI - Neanderthal DNA yields to genome foray. PMID- 16710378 TI - Antibiotic faces uncertain future. PMID- 16710379 TI - How does a painkiller harm the heart? PMID- 16710380 TI - Blogger reveals China's migratory goose farms near site of flu outbreak. PMID- 16710381 TI - Economists claim carbon cuts won't break the world's bank. PMID- 16710383 TI - Top five in physics. PMID- 16710384 TI - Who is Mark Myers? PMID- 16710386 TI - Visual neuroscience: look and learn. PMID- 16710387 TI - Microbiology: batteries not included. What can't bacteria do? PMID- 16710388 TI - Microbiology: batteries not included. Circuits of slime. PMID- 16710389 TI - Japanese spin-offs face struggle for survival. PMID- 16710392 TI - Supporting the use of artemisinin in combination. PMID- 16710393 TI - ID paper wasn't impartial enough for committee. PMID- 16710394 TI - Drug giants hamstrung by timid middle management. PMID- 16710395 TI - TGN1412: scrutinizing preclinical trials of antibody-based medicines. PMID- 16710396 TI - Neglected tests for neglected patients. PMID- 16710401 TI - Time for a change. PMID- 16710402 TI - Sociobiology: the Phoenix effect. PMID- 16710403 TI - Extrasolar planets: a neptunian triplet. PMID- 16710405 TI - Solid-state physics: when is a metal not a metal? PMID- 16710404 TI - Microbiology: antibiotic stops 'ping-pong' match. PMID- 16710407 TI - Plasma physics: cool vibes. PMID- 16710406 TI - Cell biology: skin care by keratins. PMID- 16710408 TI - Evolution: experiments in botany. PMID- 16710409 TI - Device physics: a bug-beating diode. PMID- 16710410 TI - Microbial sciences: the superficial life of microbes. PMID- 16710411 TI - Language evolution: semantic combinations in primate calls. AB - Syntax sets human language apart from other natural communication systems, although its evolutionary origins are obscure. Here we show that free-ranging putty-nosed monkeys combine two vocalizations into different call sequences that are linked to specific external events, such as the presence of a predator and the imminent movement of the group. Our findings indicate that non-human primates can combine calls into higher-order sequences that have a particular meaning. PMID- 16710412 TI - An extrasolar planetary system with three Neptune-mass planets. AB - Over the past two years, the search for low-mass extrasolar planets has led to the detection of seven so-called 'hot Neptunes' or 'super-Earths' around Sun-like stars. These planets have masses 5-20 times larger than the Earth and are mainly found on close-in orbits with periods of 2-15 days. Here we report a system of three Neptune-mass planets with periods of 8.67, 31.6 and 197 days, orbiting the nearby star HD 69830. This star was already known to show an infrared excess possibly caused by an asteroid belt within 1 au (the Sun-Earth distance). Simulations show that the system is in a dynamically stable configuration. Theoretical calculations favour a mainly rocky composition for both inner planets, while the outer planet probably has a significant gaseous envelope surrounding its rocky/icy core; the outer planet orbits within the habitable zone of this star. PMID- 16710413 TI - Evolution of an obligate social cheater to a superior cooperator. AB - Obligate relationships have evolved many times and can be parasitic or mutualistic. Obligate organisms rely on others to survive and thus coevolve with their host or partner. An important but little explored question is whether obligate status is an evolutionarily terminal condition or whether obligate lineages can evolve back to an autonomous lifestyle. The bacterium Myxococcus xanthus survives starvation by the social development of spore-bearing fruiting bodies. Some M. xanthus genotypes defective at fruiting body development in isolation can nonetheless exploit proficient genotypes in chimaeric groups. Here we report an evolutionary transition from obligate dependence on an altruistic host to an autonomous mode of social cooperation. This restoration of social independence was caused by a single mutation of large effect that confers fitness superiority over both ancestral genotypes, including immunity from exploitation by the ancestral cheater. Thus, a temporary state of obligate cheating served as an evolutionary stepping-stone to a novel state of autonomous social dominance. PMID- 16710415 TI - Suppression of dwarf galaxy formation by cosmic reionization. AB - A large number of faint galaxies, born less than a billion years after the Big Bang, have recently been discovered. Fluctuations in the distribution of these galaxies contributed to a scatter in the ionization fraction of cosmic hydrogen on scales of tens of megaparsecs, as observed along the lines of sight to the earliest known quasars. Theoretical simulations predict that the formation of dwarf galaxies should have been suppressed after cosmic hydrogen was reionized, leading to a drop in the cosmic star-formation rate. Here we report evidence for this suppression. We show that the post-reionization galaxies that produced most of the ionizing radiation at a redshift z approximately 5.5 must have had a mass in excess of approximately 10(10.9 +/- 0.5) solar masses (M(o)) or else the aforementioned scatter would have been smaller than observed. This limiting mass is two orders of magnitude larger than the galaxy mass that is thought to have dominated the reionization of cosmic hydrogen (approximately 10(8) M(o)). We predict that future surveys with space-based infrared telescopes will detect a population of smaller galaxies that reionized the Universe at an earlier time, before the epoch of dwarf galaxy suppression. PMID- 16710416 TI - An aluminium nitride light-emitting diode with a wavelength of 210 nanometres. AB - Compact high-efficiency ultraviolet solid-state light sources--such as light emitting diodes (LEDs) and laser diodes--are of considerable technological interest as alternatives to large, toxic, low-efficiency gas lasers and mercury lamps. Microelectronic fabrication technologies and the environmental sciences both require light sources with shorter emission wavelengths: the former for improved resolution in photolithography and the latter for sensors that can detect minute hazardous particles. In addition, ultraviolet solid-state light sources are also attracting attention for potential applications in high-density optical data storage, biomedical research, water and air purification, and sterilization. Wide-bandgap materials, such as diamond and III-V nitride semiconductors (GaN, AlGaN and AlN; refs 3-10), are potential materials for ultraviolet LEDs and laser diodes, but suffer from difficulties in controlling electrical conduction. Here we report the successful control of both n-type and p type doping in aluminium nitride (AlN), which has a very wide direct bandgap of 6 eV. This doping strategy allows us to develop an AlN PIN (p-type/intrinsic/n type) homojunction LED with an emission wavelength of 210 nm, which is the shortest reported to date for any kind of LED. The emission is attributed to an exciton transition, and represents an important step towards achieving exciton related light-emitting devices as well as replacing gas light sources with solid state light sources. PMID- 16710417 TI - Links between annual, Milankovitch and continuum temperature variability. AB - Climate variability exists at all timescales-and climatic processes are intimately coupled, so that understanding variability at any one timescale requires some understanding of the whole. Records of the Earth's surface temperature illustrate this interdependence, having a continuum of variability following a power-law scaling. But although specific modes of interannual variability are relatively well understood, the general controls on continuum variability are uncertain and usually described as purely stochastic processes. Here we show that power-law relationships of surface temperature variability scale with annual and Milankovitch-period (23,000- and 41,000-year) cycles. The annual cycle corresponds to scaling at monthly to decadal periods, while millennial and longer periods are tied to the Milankovitch cycles. Thus the annual, Milankovitch and continuum temperature variability together represent the response to deterministic insolation forcing. The identification of a deterministic control on the continuum provides insight into the mechanisms governing interannual and longer-period climate variability. PMID- 16710418 TI - Seismic detection of folded, subducted lithosphere at the core-mantle boundary. AB - Seismic tomography has been used to infer that some descending slabs of oceanic lithosphere plunge deep into the Earth's lower mantle. The fate of these slabs has remained unresolved, but it has been postulated that their ultimate destination is the lowermost few hundred kilometres of the mantle, known as the D'' region. Relatively cold slab material may account for high seismic velocities imaged in D'' beneath areas of long-lived plate subduction, and for reflections from a seismic velocity discontinuity just above the anomalously high wave speed regions. The D'' discontinuity itself is probably the result of a phase change in relatively low-temperature magnesium silicate perovskite. Here, we present images of the D'' region beneath the Cocos plate using Kirchhoff migration of horizontally polarized shear waves, and find a 100-km vertical step occurring over less than 100 km laterally in an otherwise flat D'' shear velocity discontinuity. Folding and piling of a cold slab that has reached the core-mantle boundary, as observed in numerical and experimental models, can account for the step by a 100-km elevation of the post-perovskite phase boundary due to a 700 degrees C lateral temperature reduction in the folded slab. We detect localized low velocities at the edge of the slab material, which may result from upwellings caused by the slab laterally displacing a thin hot thermal boundary layer. PMID- 16710414 TI - The DNA sequence and biological annotation of human chromosome 1. AB - The reference sequence for each human chromosome provides the framework for understanding genome function, variation and evolution. Here we report the finished sequence and biological annotation of human chromosome 1. Chromosome 1 is gene-dense, with 3,141 genes and 991 pseudogenes, and many coding sequences overlap. Rearrangements and mutations of chromosome 1 are prevalent in cancer and many other diseases. Patterns of sequence variation reveal signals of recent selection in specific genes that may contribute to human fitness, and also in regions where no function is evident. Fine-scale recombination occurs in hotspots of varying intensity along the sequence, and is enriched near genes. These and other studies of human biology and disease encoded within chromosome 1 are made possible with the highly accurate annotated sequence, as part of the completed set of chromosome sequences that comprise the reference human genome. PMID- 16710419 TI - Embryological evidence for developmental lability during early angiosperm evolution. AB - Recent advances in angiosperm phylogeny reconstruction, palaeobotany and comparative organismic biology have provided the impetus for a major re evaluation of the earliest phases of the diversification of flowering plants. We now know that within the first fifteen million years of angiosperm history, three major lineages of flowering plants-monocotyledons, eumagnoliids and eudicotyledons-were established, and that within this window of time, tremendous variation in vegetative and floral characteristics evolved. Here I report on a novel type of embryo sac (angiosperm female gametophyte or haploid egg-producing structure) in Amborella trichopoda, the sole member of the most ancient extant angiosperm lineage. This is the first new pattern of embryo sac structure to be discovered among angiosperms in well over half a century. This discovery also supports the emerging view that the earliest phases of angiosperm evolution were characterized by an extensive degree of developmental experimentation and structural lability, and may provide evidence of a critical link to the gymnospermous ancestors of flowering plants. PMID- 16710420 TI - Horizontal endosymbiont transmission in hydrothermal vent tubeworms. AB - Transmission of obligate bacterial symbionts between generations is vital for the survival of the host. Although the larvae of certain hydrothermal vent tubeworms (Vestimentifera, Siboglinidae) are symbiont-free and possess a transient digestive system, these structures are lost during development, resulting in adult animals that are nutritionally dependent on their bacterial symbionts. Thus, each generation of tubeworms must be newly colonized with its specific symbiont. Here we present a model for tubeworm symbiont acquisition and the development of the symbiont-housing organ, the trophosome. Our data indicate that the bacterial symbionts colonize the developing tube of the settled larvae and enter the host through the skin, a process that continues through the early juvenile stages during which the trophosome is established from mesodermal tissue. In later juvenile stages we observed massive apoptosis of host epidermis, muscles and undifferentiated mesodermal tissue, which was coincident with the cessation of the colonization process. Characterizing the symbiont transmission process in this finely tuned mutualistic symbiosis provides another model of symbiont acquisition and additional insights into underlying mechanisms common to both pathogenic infections and beneficial host-symbiont interactions. PMID- 16710422 TI - A keratin cytoskeletal protein regulates protein synthesis and epithelial cell growth. AB - Cell growth, an increase in mass and size, is a highly regulated cellular event. The Akt/mTOR (mammalian target of rapamycin) signalling pathway has a central role in the control of protein synthesis and thus the growth of cells, tissues and organisms. A striking example of a physiological context requiring rapid cell growth is tissue repair in response to injury. Here we show that keratin 17, an intermediate filament protein rapidly induced in wounded stratified epithelia, regulates cell growth through binding to the adaptor protein 14-3-3sigma. Mouse skin keratinocytes lacking keratin 17 (ref. 4) show depressed protein translation and are of smaller size, correlating with decreased Akt/mTOR signalling activity. Other signalling kinases have normal activity, pointing to the specificity of this defect. Two amino acid residues located in the amino-terminal head domain of keratin 17 are required for the serum-dependent relocalization of 14-3-3sigma from the nucleus to the cytoplasm, and for the concomitant stimulation of mTOR activity and cell growth. These findings reveal a new and unexpected role for the intermediate filament cytoskeleton in influencing cell growth and size by regulating protein synthesis. PMID- 16710421 TI - Platensimycin is a selective FabF inhibitor with potent antibiotic properties. AB - Bacterial infection remains a serious threat to human lives because of emerging resistance to existing antibiotics. Although the scientific community has avidly pursued the discovery of new antibiotics that interact with new targets, these efforts have met with limited success since the early 1960s. Here we report the discovery of platensimycin, a previously unknown class of antibiotics produced by Streptomyces platensis. Platensimycin demonstrates strong, broad-spectrum Gram positive antibacterial activity by selectively inhibiting cellular lipid biosynthesis. We show that this anti-bacterial effect is exerted through the selective targeting of beta-ketoacyl-(acyl-carrier-protein (ACP)) synthase I/II (FabF/B) in the synthetic pathway of fatty acids. Direct binding assays show that platensimycin interacts specifically with the acyl-enzyme intermediate of the target protein, and X-ray crystallographic studies reveal that a specific conformational change that occurs on acylation must take place before the inhibitor can bind. Treatment with platensimycin eradicates Staphylococcus aureus infection in mice. Because of its unique mode of action, platensimycin shows no cross-resistance to other key antibiotic-resistant strains tested, including methicillin-resistant S. aureus, vancomycin-intermediate S. aureus and vancomycin resistant enterococci. Platensimycin is the most potent inhibitor reported for the FabF/B condensing enzymes, and is the only inhibitor of these targets that shows broad-spectrum activity, in vivo efficacy and no observed toxicity. PMID- 16710423 TI - The mechanism of cell differentiation in Bacillus subtilis. AB - Sporulation in Bacillus subtilis serves as a model for the development of two different cell types from a single cell. Although much information has been accumulated about the mechanisms that initiate the developmental programmes, important questions remain that can be answered only by quantitative analysis. Here we develop, with the help of existing and new experimental results, a mathematical model that reproduces published in vitro experiments and explains how the activation of the key transcription factor is regulated. The model identifies the difference in volume between the two cell types as the primary trigger for determining cell fate. It shows that this effect depends on the allosteric behaviour of a key protein kinase and on a low rate of dephosphorylation by the corresponding phosphatase; both predicted effects are confirmed experimentally. PMID- 16710424 TI - Cleavage of pre-tRNAs by the splicing endonuclease requires a composite active site. AB - Splicing is required for the removal of introns from a subset of transfer RNAs in all eukaryotic organisms. The first step of splicing, intron recognition and cleavage, is performed by the tRNA-splicing endonuclease, a tetrameric enzyme composed of the protein subunits Sen54, Sen2, Sen34 and Sen15. It has previously been demonstrated that the active sites for cleavage at the 5' and 3' splice sites of precursor tRNA are contained within Sen2 and Sen34, respectively. A recent structure of an archaeal endonuclease complexed with a bulge-helix-bulge RNA has led to the unexpected hypothesis that catalysis requires a critical 'cation-pi sandwich' composed of two arginine residues that serve to position the RNA substrate within the active site. This motif is derived from a cross-subunit interaction between the two catalytic subunits. Here we test the role of this interaction within the eukaryotic endonuclease and show that catalysis at the 5' splice site requires the conserved cation-pi sandwich derived from the Sen34 subunit in addition to the catalytic triad of Sen2. The catalysis of pre-tRNA by the eukaryotic tRNA-splicing endonuclease therefore requires a previously unrecognized composite active site. PMID- 16710426 TI - Hospital Episode Statistics and changing trends in glaucoma surgery. AB - BACKGROUND: A number of authors have documented the decrease in the rate of glaucoma drainage surgery, felt to be owing to increased medical therapy options. There has also been an increase in cataract extraction. The aim of this study was, using the NHS Hospital Episode Statistics (HES), to attempt to confirm these trends and to examine the possibility that these trends are not necessarily independent. METHODS: The HES was accessed for the main glaucoma procedures between 1998 and 2004 and for cataract operations performed over the same time period. Diagnostic data were also extracted from the HES data for glaucoma. Figures for the changes in glaucoma medications were obtained from pharmaceutical data. RESULTS: Over the period that the data are available, trabeculectomy numbers have reduced by 51%, laser trabeculoplasty by 60%, and laser peripheral iridectomy (PI) by 30%. Cataract extractions have increased by 52%. Medical treatments, especially prostaglandin use, have increased. The number of admissions for acute glaucoma did not increase. CONCLUSIONS: The figures indicated a decrease in glaucoma drainage surgery and an increase in cataract extraction-consistent with other studies. More surprisingly, there was no increase in diagnosis of angle closure glaucoma or laser PIs within the figures and we suggest this may in part be related to the increase in cataract extraction. We speculate that this may also partly be responsible for the reduction in trabeculectomies. Although the HES data are a potentially rich source of information, there are potential inaccuracies in the data, which means interpretations must be made with caution. PMID- 16710427 TI - Flow cytometric analysis of HLA-DR antigen in conjunctival epithelial cells of patients with cystic fibrosis. AB - PURPOSE: Cystic fibrosis (CF) is an autosomal-recessive genetic disorder. The disease affects all secretory epithelia including the eye and belongs to the group of ocular surface epithelial diseases, termed keratoconjunctivitis sicca that develop in dry eye. In the pathogenesis of dry eye, inflammation plays a crucial role. The aim of this study was to investigate the expression of HLA-DR on conjunctival epithelial cells from patients with CF. MATERIALS AND METHODS: Twenty-five patients with CF and 25 normal subjects underwent ocular examination. Tear film break-up time (TBUT), Schirmer test, lissamine green staining, and conjunctival impression cytology were carried out. Cells were processed for flow cytometry, by using monoclonal antibodies to HLA-DR. RESULTS: The Schirmer test and TBUT scores were significantly lower in CF patients compared with controls. A significant increase of HLA-DR expression on epithelial cells was found in patients with CF compared with normal eyes. The Schirmer and TBUT test were positively correlated with HLA-DR expression for the percentage of cells. CONCLUSION: These results suggest that conjunctival epithelial cells play an important proinflammatory role in ocular changes in CF patients. Our findings confirm the presence of an inflammatory background and the immune nature of this disease. HLA-DR measurement might be a useful method for monitoring of inflammatory processes in the conjunctiva and could be helpful in the use of anti inflammatory drugs in the treatment of ocular findings in CF patients. PMID- 16710428 TI - Current opinion on treatment of asymptomatic retinal detachments. AB - AIMS: To determine the current opinion of vitreoretinal consultants in the UK on the treatment of asymptomatic retinal detachments. METHODS: A questionnaire survey of UK and Eire vitreoretinal consultants on recommended therapy for eight different case scenarios of asymptomatic retinal detachment. RESULTS: A questionnaire was sent to 115 consultants in UK and Eire. Fifty-four per cent were returned. The majority of surgeons favoured active intervention for most types of asymptomatic retinal detachment, with surgery being the most popular. Laser demarcation of the detached retina was the second most favoured approach followed by observation alone. Discharge with advice to seek attention if symptoms appeared was the least frequently recommended. Recommended management varied considerably for each scenario. The presence or absence of a demarcation line had the greatest influence on the management of each case. Age of patient had a much smaller effect, and degree of myopia had very little effect. CONCLUSIONS: Surgery remains the most common recommended management for an asymptomatic retinal detachment. The striking outcome of the survey is the variability in responses for any given scenario, ranging from surgical intervention to the patient being discharged with advice. This reflects the absence of hard clinical evidence to support any particular management option in such cases. As patient involvement in decision making over management is essential, the opinions expressed in this survey may not be a true reflection of actual practice, although it does highlight the need for further research in this field to provide guidance for treatment of this important condition. PMID- 16710429 TI - Cystoid macular oedema following pneumatic retinopexy vs scleral buckling. AB - AIMS: To determine the incidence of angiographic cystoid macular oedema (CMO) following pneumatic retinopexy (PR) and scleral buckling (SB) in consecutive case series. METHODS: Patients who had successful anatomical attachment following PR and SB were included in our study; 132 patients had PR and 121 patients had SB. We evaluated the demographic characteristics, visual acuity, lens status, macular status, and previous ocular history in all patients. CMO was evaluated by fluorescein angiography (FA) in a masked pattern, 6 and 12 weeks after surgery in all cases. We analysed the CMO incidence and its correlation with preoperative ocular status and visual outcome. Chi2 and Fisher's exact tests were used in statistical analysis. RESULTS: CMO was present in 15 of 132 (11%) PR, and 35 of 121 (29%) SB patients at 6 weeks (P=0.0005); the oedema was persistent in eight of 132 (6%) PR and 21 of 121 (17%) SB patients at 12 weeks (P=0.0005). Eight of 106 (8%) phakic and seven of 26 (27%) pseudophakic patients developed CMO following PR (P=0.02). In the SB group, 26 of 72 (36%) patients who had preoperative macular detachment developed CMO (P=0.03). Visual improvement was limited in patients who developed angiographic CMO despite anatomical re attachment of the retina. CONCLUSIONS: CMO may occur following both PR and SB and deteriorate the visual outcome. Previous cataract surgery and macular detachment may increase the CMO rates following PR and SB, respectively. PMID- 16710430 TI - Ocular coherence tomographic examination of postoperative foveal architecture after scleral buckling vs vitrectomy for macular off retinal detachment. AB - AIMS: This pilot study uses Optical Coherence Tomography (OCT) imaging to compare the difference in foveal architecture after successful retinal detachment (RD) surgery by scleral buckling or pars plana vitrectomy (PPV). METHODS: Prospective recruitment of patients with macular off RDs. Detachment surgery was undertaken by scleral buckling, external drainage, and air injection (group 1) or by PPV (group 2). Postoperatively patients had clinical examinations and OCT at 1, 3, 6, and 12 months. If abnormalities persisted, a further OCT was obtained at 18 months. RESULTS: Retinal reattachment, including clinical macular reattachment, was achieved in all cases within 24 h postoperatively. In group 1 (n=22), postoperative OCT showed persistent foveal detachment in 63% of cases (n=14) at 1 and 3 months. At 6 and 12 months, 36% (n=8) and 9% (n=2) had a persistent foveal detachment, respectively, and at 18 months, foveal detachment eventually. In group 2 (n=21), postoperative OCT showed an attached fovea in all cases; however, foveal thickening suggesting intraretinal oedema was present in all cases. The oedematous appearance of retina on OCT settled in 1-3 months. No foveal abnormality was seen at 6 and 12 months postoperatively. CONCLUSIONS: A high proportion of patients with successful retinal reattachment surgery by scleral buckling had foveal detachments postoperatively. No cases who had PPV had foveal detachments; however, transient retinal oedema was evident in all cases. The aetiology of these changes is unknown and warrants further investigation, as there is the potential of a long-term effect on vision. PMID- 16710431 TI - Patients' expectation and experience of visual sensations during phacoemulsification under topical anaesthesia. AB - PURPOSE: To describe what patients expect to see and the visual sensations they actually experience during phacoemulsification under topical anaesthesia. We also sought to determine if patients find their intraoperative visual experience frightening and the factors associated with this. MATERIALS AND METHODS: Ninety eight patients who underwent phacoemulsification and intraocular lens implantation under topical anaesthesia were interviewed preoperatively on what they expected to see with their operated eye during surgery and again postoperatively on what they actually saw. No patient received counselling about possible intraoperative visual sensations. A logistic (multivariate) regression model was used for statistical analysis. RESULTS: Preoperatively, 36 patients (36.7%) expected at least light perception, 38 (38.8%) expected no light perception, and 24 (24.5%) were unsure what to expect. Some patients also expected a variety of different visual sensations. Postoperatively, all patients (100%) reported seeing light intraoperatively and many experienced various other visual sensations. Nineteen patients (19.4%) found their visual experience frightening. The following factors were statistically associated with a frightening visual experience: preoperative anxiety, previous cataract surgery in the fellow eye, experiencing an intraoperative increase in clarity, not seeing movement intraoperatively, and not knowing what to expect. CONCLUSIONS: The majority of patients in this study either expected that they would see nothing at all during the surgery or were unsure of what to expect. All patients subsequently saw at least some light, and many perceived various other visual sensations that were frightening to nearly one in five patients. Preoperative counselling should inform about possible intraoperative visual experience. PMID- 16710432 TI - Successful treatment of macular holes associated with peripheral retinal vascular tumours. AB - BACKGROUND: Macular holes have been described in association with benign vascular tumours of the retina. However, their successful treatment has not been reported. PATIENTS AND METHODS: Three patients with macular holes associated with retinal vascular tumours had vitrectomy, peeling of the internal limiting membrane, and intraocular gas tamponade. RESULTS: In all three cases, the macular holes were successfully closed, and the visual acuity improved. CONCLUSION: Macular holes associated with benign retinal vascular tumours respond to conventional macular hole surgical techniques. PMID- 16710433 TI - Ocular morbidity associated with intravitreal triamcinolone acetonide. AB - AIM: To report on the complications associated with the use of intravitreal triamcinolone acetonide (IVTA) in a tertiary referral hospital setting. MATERIALS AND METHODS: A retrospective case series review of all IVTA injections carried out over a period of 30 months. RESULTS: One hundred and thirty IVTA injections were performed; nine with limited local follow-up were excluded. Thus, 121 injections (108 patients, 114 eyes) were included in the study. Triamcinolone (4 mg) was used in all cases. Indications were diabetic macular oedema (n=41 eyes), retinal vein occlusions (n=27), postoperative cystoid macular oedema (n=24), exudative age-related macular degeneration (n=16), and others (n=6). No intraoperative complications were recorded. Postoperative intraocular pressure (IOP) readings of 22, 28, 35, and 40 mmHg or higher were recorded in 46.5, 29.8, 12.3, and 7.9% of eyes, respectively. IOP elevation was treated with antiglaucoma medication in all but one eye (0.9%) that required trabeculectomy and one (0.9%) that required vitrectomy with cataract extraction for suspected phacoanaphylactic glaucoma. Two eyes (1.8%) developed retinal detachment; both had previously been treated for retinal breaks. One eye (0.9%) developed culture-positive endophthalmitis. CONCLUSIONS: Significant morbidity is associated with IVTA injection; clinicians should be aware when considering treatment options. PMID- 16710434 TI - Lamellar corneal dissection for visualization of the anterior chamber before triple procedure. AB - PURPOSE: Open-sky cataract extraction during triple procedure can be associated with higher risk of complications owing to the missing counterbalance by the cornea. Herein, we present a fast and easy technique for visualization of the anterior chamber and the lens in eyes with opaque corneas planed for triple procedure. MATERIALS AND METHODS: Four patients with corneal oedema due to Fuchs' endothelial dystrophy and cataract underwent triple procedure. As the anterior chamber view was limited, the central 7.0 mm of the cornea was marked. Then, 60 80% of the corneal thickness was removed by lamellar dissection and filled with hydroxypropylmethylcellulose. Continuous curvilinear capsulorhexis (CCC) and phacoemulsification with intraocular lens implantation through a corneoscleral tunnel were then performed and at the end the remaining corneal tissue was excised and the donor tissue fixed with a single running continuous suture. RESULTS: Lamellar corneal dissection enhances the anterior chamber view and CCC can be performed under stable anterior chamber condition. Phacoemulsification via sclerocorneal tunnel could be easily performed under good anterior chamber view in all cases. The operation time was 60-75 min in all cases. CONCLUSION: Lamellar corneal dissection in opaque corneas before cataract extraction is a useful technique for enhancing anterior chamber view in cases of triple procedure. PMID- 16710435 TI - A simple confocal fibre-optic laser method for intraocular lens power measurement. AB - PURPOSE: To develop novel confocal fibre-optic laser method (CFOLM) for accurate and objective measuring of the dioptric power of both positive and negative intraocular lenses (IOLs). METHODS: The CFOLM principle of operation is based on a simple apertureless single-mode fibre laser confocal design. The key element is a single-mode fibre coupler that serves simultaneously as a point light source (3 5 microm fibre diameter) used for the formation of a collimated Gaussian beam, and as a confocal point receiver that is highly sensitive to spatial displacements of the focused backreflectance laser emission. The basic CFOLM systems include IOL testing set-ups for the measurement of both positive and negative IOLs. RESULTS: The CFOLM designs provide high accuracy (or=8 mmHg rise from baseline with iridocorneal touch was considered as a positive test. RESULTS: Of the 149 family members examined, 55 (36.9%) were found to have PACG. Forty (72.7%) of these had subacute PACG and 15 (27.3%) were found to have chronic PACG. Thirty-nine (70.3%) of the affected members showed a positive DRPPT. Mean anterior chamber depth (ACD) was 2.03+/ 0.3, 2.3+/-0.4, 2.7+/-0.3 mm (P=0.0001) and mean lens thickness was 4.41+/-0.39, 3.99+/-0.5, 3.93+/-0.4 mm (P=0.0001) in DRPPT positives, borderlines and negatives respectively. ROC curve (ACD) plotted showed cutoff value of 2.07 mm (sensitivity 88.57%) for screening. CONCLUSION: Anterior chamber is shallowest, lens is thickest and axial length is shortest in affected and DRPPT positive, family members of PACG patients. PMID- 16710439 TI - Microincision bimanual phacoemulsification and Thinoptx implantation through a 1.70 mm incision. AB - PURPOSE: To prospectively assess the efficacy of bimanual phacoemulsification and implantation of Thinoptx, an injectable intraocular lens (IOL), inserted through 1.70 mm clear corneal incision. SETTING: Department of Ophthalmology, Southend Hospital NHS Trust, UK. METHODS: A total of 50 eyes of 49 randomly selected patients with cataracts had microincision clear corneal bimanual phacoemulsification (MICS) with implantation of Thinoptx IOL in the capsular bag. All patients underwent full preoperative assessment. Postoperative assessment was carried out at 3 and 6 weeks and at 15 months. RESULTS: In all 50 cases the IOL was inserted through 1.70 mm clear corneal incision. The mean best-corrected visual acuity was 0.02 (6/6-1) at 6 weeks and was 0.17 (6/10) at the final follow up. The mean final surgically induced astigmatism at 6 weeks was 0.0106. Coloured haloes around artificial lights were perceived by 69.23% of patients at 6 weeks and by 61.29% at the final follow-up. One patient underwent IOL exchange for this. Posterior capsular opacification was noticed in 31.26% at 6 weeks and in 64.51% at 15 months. Anterior capsular opacification was noticed in 5.26% at 6 weeks and in 16.12% at 15 months. In one patient the IOL had to be exchanged because of tilt and displacement of the IOL due to anterior capsular phimosis. CONCLUSIONS: We conclude Thinoptx can be safely inserted through 1.70 mm incision used for bimanual phacoemulsification. Distance and near visual acuity achieved with this IOL is satisfactory. There is no significant change in keratometric astigmatism following this procedure. However, posterior capsular opacification rate was significantly higher with this IOL. Haloes around light sources were significant. PMID- 16710438 TI - Laser-induced chorioretinal venous anastomosis using combined lasers with different wavelengths. AB - AIM: To evaluate the effectiveness of combined lasers with different wavelengths in creating chorioretinal venous anastomosis (CRVA) in nonischaemic central retinal vein occlusion (CRVO). METHODS: In this retrospective, noncomparative, consecutive case series, records of all patients with a diagnosis of nonischaemic CRVO who underwent combined laser treatment to induce CRVA and who were followed up for 6 months or longer were reviewed. RESULTS: Records of 85 patients (85 eyes) were analysed. These patients were divided into two groups. In group 1 (24 cases, 24 eyes), combined lasers with two different wavelengths (argon blue-green laser combined with Nd-YAG laser were used. In group 2 (61 cases, 61 eyes), combined lasers with three different wavelengths (argon blue-green laser combined with krypton red laser and Nd-YAG laser) were used. Functional anastomosis formed in 13 out of 24 eyes (54%) in group 1 and 47 out of 61 eyes (77%) in group 2. In patients with successful anastomosis, visual acuity improved for two or more lines in 49 out of 60 eyes (82%), with the remaining eyes undergoing no change. There were no serious complications. CONCLUSION: The use of three laser wavelengths results in the creation of laser anastomosis and the success rate may be higher than when using two wavelengths alone. PMID- 16710440 TI - Femtosecond Time-Resolved Stimulated Raman Spectroscopy: Application to the Ultrafast Internal Conversion in beta-Carotene. AB - We have developed the technique of femtosecond stimulated Raman spectroscopy (FSRS), which allows the rapid collection of high-resolution vibrational spectra on the femtosecond time scale. FSRS combines a sub-50 fs actinic pump pulse with a two-pulse stimulated Raman probe to obtain vibrational spectra whose frequency resolution limits are uncoupled from the time resolution. This allows the acquisition of spectra with <100 fs time resolution and <30 cm(-1) frequency resolution. Additionally, FSRS is unaffected by background fluorescence, provides rapid (100 ms) acquisition times, and exhibits traditional spontaneous Raman line shapes. FSRS is used here to study the relaxation dynamics of beta-carotene. Following optical excitation to S(2) (1B(u) (+)) the molecule relaxes in 160 fs to S(1) (2A(g) (-)) and then undergoes two distinct stages of intramolecular vibrational energy redistribution (IVR) with 200 and 450 fs time constants. These processes are attributed to rapid (200 fs) distribution of the internal conversion energy from the S(1) C=C modes into a restricted bath of anharmonically coupled modes followed by complete IVR in 450 fs. FSRS is a valuable new technique for studying the vibrational structure of chemical reaction intermediates and transition states. PMID- 16710442 TI - Stereoselective construction of cis-2,6-disubstituted tetrahydropyrans via an intramolecular bismuth-mediated oxa-conjugate addition reaction. AB - The intramolecular oxa-conjugate addition of tethered triethylsilyloxy substituted alpha,beta-unsaturated ketones mediated by bismuth(III) nitrate pentahydrate provides a mild and efficient method for the stereoselective construction of cis-2,6-disubstituted tetrahydropyrans. PMID- 16710441 TI - Detection of temporal changes in the spatial distribution of cancer rates using local Moran's I and geostatistically simulated spatial neutral models. AB - This paper presents the first application of spatially correlated neutral models to the detection of changes in mortality rates across space and time using the local Moran's I statistic. Sequential Gaussian simulation is used to generate realizations of the spatial distribution of mortality rates under increasingly stringent conditions: 1) reproduction of the sample histogram, 2) reproduction of the pattern of spatial autocorrelation modeled from the data, 3) incorporation of regional background obtained by geostatistical smoothing of observed mortality rates, and 4) incorporation of smooth regional background observed at a prior time interval. The simulated neutral models are then processed using two new spatio-temporal variants of the Morany's I statistic, which allow one to identify significant changes in mortality rates above and beyond past spatial patterns. Last, the results are displayed using an original classification of clusters/outliers tailored to the space-time nature of the data. Using this new methodology the space-time distribution of cervix cancer mortality rates recorded over all US State Economic Areas (SEA) is explored for 9 time periods of 5 years each. Incorporation of spatial autocorrelation leads to fewer significant SEA units than obtained under the traditional assumption of spatial independence, confirming earlier claims that Type I errors may increase when tests using the assumption of independence are applied to spatially correlated data. Integration of regional background into the neutral models yields substantially different spatial clusters and outliers, highlighting local patterns which were blurred when local Moran's I was applied under the null hypothesis of constant risk. PMID- 16710444 TI - Nonparametric Methods in Reliability. AB - Probabilistic and statistical models for the occurrence of a recurrent event over time are described. These models have applicability in the reliability, engineering, biomedical and other areas where a series of events occurs for an experimental unit as time progresses. Nonparametric inference methods, in particular, the estimation of a relevant distribution function, are described. PMID- 16710445 TI - Epigenetic telomere protection by Drosophila DNA damage response pathways. AB - Analysis of terminal deletion chromosomes indicates that a sequence-independent mechanism regulates protection of Drosophila telomeres. Mutations in Drosophila DNA damage response genes such as atm/tefu, mre11, or rad50 disrupt telomere protection and localization of the telomere-associated proteins HP1 and HOAP, suggesting that recognition of chromosome ends contributes to telomere protection. However, the partial telomere protection phenotype of these mutations limits the ability to test if they act in the epigenetic telomere protection mechanism. We examined the roles of the Drosophila atm and atr-atrip DNA damage response pathways and the nbs homolog in DNA damage responses and telomere protection. As in other organisms, the atm and atr-atrip pathways act in parallel to promote telomere protection. Cells lacking both pathways exhibit severe defects in telomere protection and fail to localize the protection protein HOAP to telomeres. Drosophila nbs is required for both atm- and atr-dependent DNA damage responses and acts in these pathways during DNA repair. The telomere fusion phenotype of nbs is consistent with defects in each of these activities. Cells defective in both the atm and atr pathways were used to examine if DNA damage response pathways regulate telomere protection without affecting telomere specific sequences. In these cells, chromosome fusion sites retain telomere specific sequences, demonstrating that loss of these sequences is not responsible for loss of protection. Furthermore, terminally deleted chromosomes also fuse in these cells, directly implicating DNA damage response pathways in the epigenetic protection of telomeres. We propose that recognition of chromosome ends and recruitment of HP1 and HOAP by DNA damage response proteins is essential for the epigenetic protection of Drosophila telomeres. Given the conserved roles of DNA damage response proteins in telomere function, related mechanisms may act at the telomeres of other organisms. PMID- 16710446 TI - Genome-wide mapping of susceptibility to coronary artery disease identifies a novel replicated locus on chromosome 17. AB - Coronary artery disease (CAD) is a leading cause of death world-wide, and most cases have a complex, multifactorial aetiology that includes a substantial heritable component. Identification of new genes involved in CAD may inform pathogenesis and provide new therapeutic targets. The PROCARDIS study recruited 2,658 affected sibling pairs (ASPs) with onset of CAD before age 66 y from four European countries to map susceptibility loci for CAD. ASPs were defined as having CAD phenotype if both had CAD, or myocardial infarction (MI) phenotype if both had a MI. In a first study, involving a genome-wide linkage screen, tentative loci were mapped to Chromosomes 3 and 11 with the CAD phenotype (1,464 ASPs), and to Chromosome 17 with the MI phenotype (739 ASPs). In a second study, these loci were examined with a dense panel of grid-tightening markers in an independent set of families (1,194 CAD and 344 MI ASPs). This replication study showed a significant result on Chromosome 17 (MI phenotype; p = 0.009 after adjustment for three independent replication tests). An exclusion analysis suggests that further genes of effect size lambda(sib) > 1.24 are unlikely to exist in these populations of European ancestry. To our knowledge, this is the first genome-wide linkage analysis to map, and replicate, a CAD locus. The region on Chromosome 17 provides a compelling target within which to identify novel genes underlying CAD. Understanding the genetic aetiology of CAD may lead to novel preventative and/or therapeutic strategies. PMID- 16710447 TI - Diverse chromatin remodeling genes antagonize the Rb-involved SynMuv pathways in C. elegans. AB - In Caenorhabditis elegans, vulval cell-fate specification involves the activities of multiple signal transduction and regulatory pathways that include a receptor tyrosine kinase/Ras/mitogen-activated protein kinase pathway and synthetic multivulva (SynMuv) pathways. Many genes in the SynMuv pathways encode transcription factors including the homologs of mammalian Rb, E2F, and components of the nucleosome-remodeling deacetylase complex. To further elucidate the functions of the SynMuv genes, we performed a genome-wide RNA interference (RNAi) screen to search for genes that antagonize the SynMuv gene activities. Among those that displayed a varying degree of suppression of the SynMuv phenotype, 32 genes are potentially involved in chromatin remodeling (called SynMuv suppressor genes herein). Genetic mutations of two representative genes (zfp-1 and mes-4) were used to further characterize their positive roles in vulval induction and relationships with Ras function. Our analysis revealed antagonistic roles of the SynMuv suppressor genes and the SynMuv B genes in germline-soma distinction, RNAi, somatic transgene silencing, and tissue specific expression of pgl-1 and the lag-2/Delta genes. The opposite roles of these SynMuv B and SynMuv suppressor genes on transcriptional regulation were confirmed in somatic transgene silencing. We also report the identifications of ten new genes in the RNAi pathway and six new genes in germline silencing. Among the ten new RNAi genes, three encode homologs of proteins involved in both protein degradation and chromatin remodeling. Our findings suggest that multiple chromatin remodeling complexes are involved in regulating the expression of specific genes that play critical roles in developmental decisions. PMID- 16710448 TI - Statistics of knots, geometry of conformations, and evolution of proteins. AB - Like shoelaces, the backbones of proteins may get entangled and form knots. However, only a few knots in native proteins have been identified so far. To more quantitatively assess the rarity of knots in proteins, we make an explicit comparison between the knotting probabilities in native proteins and in random compact loops. We identify knots in proteins statistically, applying the mathematics of knot invariants to the loops obtained by complementing the protein backbone with an ensemble of random closures, and assigning a certain knot type to a given protein if and only if this knot dominates the closure statistics (which tells us that the knot is determined by the protein and not by a particular method of closure). We also examine the local fractal or geometrical properties of proteins via computational measurements of the end-to-end distance and the degree of interpenetration of its subchains. Although we did identify some rather complex knots, we show that native conformations of proteins have statistically fewer knots than random compact loops, and that the local geometrical properties, such as the crumpled character of the conformations at a certain range of scales, are consistent with the rarity of knots. From these, we may conclude that the known "protein universe" (set of native conformations) avoids knots. However, the precise reason for this is unknown--for instance, if knots were removed by evolution due to their unfavorable effect on protein folding or function or due to some other unidentified property of protein evolution. PMID- 16710449 TI - Reverse engineering the gap gene network of Drosophila melanogaster. AB - A fundamental problem in functional genomics is to determine the structure and dynamics of genetic networks based on expression data. We describe a new strategy for solving this problem and apply it to recently published data on early Drosophila melanogaster development. Our method is orders of magnitude faster than current fitting methods and allows us to fit different types of rules for expressing regulatory relationships. Specifically, we use our approach to fit models using a smooth nonlinear formalism for modeling gene regulation (gene circuits) as well as models using logical rules based on activation and repression thresholds for transcription factors. Our technique also allows us to infer regulatory relationships de novo or to test network structures suggested by the literature. We fit a series of models to test several outstanding questions about gap gene regulation, including regulation of and by hunchback and the role of autoactivation. Based on our modeling results and validation against the experimental literature, we propose a revised network structure for the gap gene system. Interestingly, some relationships in standard textbook models of gap gene regulation appear to be unnecessary for or even inconsistent with the details of gap gene expression during wild-type development. PMID- 16710450 TI - Iterative reconstruction of transcriptional regulatory networks: an algorithmic approach. AB - The number of complete, publicly available genome sequences is now greater than 200, and this number is expected to rapidly grow in the near future as metagenomic and environmental sequencing efforts escalate and the cost of sequencing drops. In order to make use of this data for understanding particular organisms and for discerning general principles about how organisms function, it will be necessary to reconstruct their various biochemical reaction networks. Principal among these will be transcriptional regulatory networks. Given the physical and logical complexity of these networks, the various sources of (often noisy) data that can be utilized for their elucidation, the monetary costs involved, and the huge number of potential experiments approximately 10(12)) that can be performed, experiment design algorithms will be necessary for synthesizing the various computational and experimental data to maximize the efficiency of regulatory network reconstruction. This paper presents an algorithm for experimental design to systematically and efficiently reconstruct transcriptional regulatory networks. It is meant to be applied iteratively in conjunction with an experimental laboratory component. The algorithm is presented here in the context of reconstructing transcriptional regulation for metabolism in Escherichia coli, and, through a retrospective analysis with previously performed experiments, we show that the produced experiment designs conform to how a human would design experiments. The algorithm is able to utilize probability estimates based on a wide range of computational and experimental sources to suggest experiments with the highest potential of discovering the greatest amount of new regulatory knowledge. PMID- 16710451 TI - Predicting Shine-Dalgarno sequence locations exposes genome annotation errors. AB - In prokaryotes, Shine-Dalgarno (SD) sequences, nucleotides upstream from start codons on messenger RNAs (mRNAs) that are complementary to ribosomal RNA (rRNA), facilitate the initiation of protein synthesis. The location of SD sequences relative to start codons and the stability of the hybridization between the mRNA and the rRNA correlate with the rate of synthesis. Thus, accurate characterization of SD sequences enhances our understanding of how an organism's transcriptome relates to its cellular proteome. We implemented the Individual Nearest Neighbor Hydrogen Bond model for oligo-oligo hybridization and created a new metric, relative spacing (RS), to identify both the location and the hybridization potential of SD sequences by simulating the binding between mRNAs and single-stranded 16S rRNA 3' tails. In 18 prokaryote genomes, we identified 2,420 genes out of 58,550 where the strongest binding in the translation initiation region included the start codon, deviating from the expected location for the SD sequence of five to ten bases upstream. We designated these as RS+1 genes. Additional analysis uncovered an unusual bias of the start codon in that the majority of the RS+1 genes used GUG, not AUG. Furthermore, of the 624 RS+1 genes whose SD sequence was associated with a free energy release of less than 8.4 kcal/mol (strong RS+1 genes), 384 were within 12 nucleotides upstream of in frame initiation codons. The most likely explanation for the unexpected location of the SD sequence for these 384 genes is mis-annotation of the start codon. In this way, the new RS metric provides an improved method for gene sequence annotation. The remaining strong RS+1 genes appear to have their SD sequences in an unexpected location that includes the start codon. Thus, our RS metric provides a new way to explore the role of rRNA-mRNA nucleotide hybridization in translation initiation. PMID- 16710452 TI - Effect of DNA repair protein Rad18 on viral infection. AB - Host factors belonging to the DNA repair machineries are assumed to aid retroviruses in the obligatory step of integration. Here we describe the effect of DNA repair molecule Rad18, a component of the post-replication repair pathway, on viral infection. Contrary to our expectations, cells lacking Rad18 were consistently more permissive to viral transduction as compared to Rad18(+/+) controls. Remarkably, such susceptibility was integration independent, since retroviruses devoid of integration activity also showed enhancement of the initial steps of infection. Moreover, the elevated sensitivity of the Rad18(-/-) cells was also observed with adenovirus. These data indicate that Rad18 suppresses viral infection in a non-specific fashion, probably by targeting incoming DNA. Furthermore, considering data published recently, it appears that the interactions between DNA repair components with incoming viruses, often result in inhibition of the infection rather than cooperation toward its establishment. PMID- 16710453 TI - The tripartite associations between bacteriophage, Wolbachia, and arthropods. AB - By manipulating arthropod reproduction worldwide, the heritable endosymbiont Wolbachia has spread to pandemic levels. Little is known about the microbial basis of cytoplasmic incompatibility (CI) except that bacterial densities and percentages of infected sperm cysts associate with incompatibility strength. The recent discovery of a temperate bacteriophage (WO-B) of Wolbachia containing ankyrin-encoding genes and virulence factors has led to intensifying debate that bacteriophage WO-B induces CI. However, current hypotheses have not considered the separate roles that lytic and lysogenic phage might have on bacterial fitness and phenotype. Here we describe a set of quantitative approaches to characterize phage densities and its associations with bacterial densities and CI. We enumerated genome copy number of phage WO-B and Wolbachia and CI penetrance in supergroup A- and B-infected males of the parasitoid wasp Nasonia vitripennis. We report several findings: (1) variability in CI strength for A-infected males is positively associated with bacterial densities, as expected under the bacterial density model of CI, (2) phage and bacterial densities have a significant inverse association, as expected for an active lytic infection, and (3) CI strength and phage densities are inversely related in A-infected males; similarly, males expressing incomplete CI have significantly higher phage densities than males expressing complete CI. Ultrastructural analyses indicate that approximately 12% of the A Wolbachia have phage particles, and aggregations of these particles can putatively occur outside the Wolbachia cell. Physical interactions were observed between approximately 16% of the Wolbachia cells and spermatid tails. The results support a low to moderate frequency of lytic development in Wolbachia and an overall negative density relationship between bacteriophage and Wolbachia. The findings motivate a novel phage density model of CI in which lytic phage repress Wolbachia densities and therefore reproductive parasitism. We conclude that phage, Wolbachia, and arthropods form a tripartite symbiotic association in which all three are integral to understanding the biology of this widespread endosymbiosis. Clarifying the roles of lytic and lysogenic phage development in Wolbachia biology will effectively structure inquiries into this research topic. PMID- 16710454 TI - Recruitment of BAD by the Chlamydia trachomatis vacuole correlates with host-cell survival. AB - Chlamydiae replicate intracellularly in a vacuole called an inclusion. Chlamydial infected host cells are protected from mitochondrion-dependent apoptosis, partly due to degradation of BH3-only proteins. The host-cell adapter protein 14-3-3beta can interact with host-cell apoptotic signaling pathways in a phosphorylation dependent manner. In Chlamydia trachomatis-infected cells, 14-3-3beta co localizes to the inclusion via direct interaction with a C. trachomatis-encoded inclusion membrane protein. We therefore explored the possibility that the phosphatidylinositol-3 kinase (PI3K) pathway may contribute to resistance of infected cells to apoptosis. We found that inhibition of PI3K renders C. trachomatis-infected cells sensitive to staurosporine-induced apoptosis, which is accompanied by mitochondrial cytochrome c release. 14-3-3beta does not associate with the Chlamydia pneumoniae inclusion, and inhibition of PI3K does not affect protection against apoptosis of C. pneumoniae-infected cells. In C. trachomatis infected cells, the PI3K pathway activates AKT/protein kinase B, which leads to maintenance of the pro-apoptotic protein BAD in a phosphorylated state. Phosphorylated BAD is sequestered via 14-3-3beta to the inclusion, but it is released when PI3K is inhibited. Depletion of AKT through short-interfering RNA reverses the resistance to apoptosis of C. trachomatis-infected cells. BAD phosphorylation is not maintained and it is not recruited to the inclusion of Chlamydia muridarum, which protects poorly against apoptosis. Thus, sequestration of BAD away from mitochondria provides C. trachomatis with a mechanism to protect the host cell from apoptosis via the interaction of a C. trachomatis-encoded inclusion protein with a host-cell phosphoserine-binding protein. PMID- 16710455 TI - Legionella pneumophila exploits PI(4)P to anchor secreted effector proteins to the replicative vacuole. AB - The causative agent of Legionnaires' disease, Legionella pneumophila, employs the intracellular multiplication (Icm)/defective organelle trafficking (Dot) type IV secretion system (T4SS) to upregulate phagocytosis and to establish a replicative vacuole in amoebae and macrophages. Legionella-containing vacuoles (LCVs) do not fuse with endosomes but recruit early secretory vesicles. Here we analyze the role of host cell phosphoinositide (PI) metabolism during uptake and intracellular replication of L. pneumophila. Genetic and pharmacological evidence suggests that class I phosphatidylinositol(3) kinases (PI3Ks) are dispensable for phagocytosis of wild-type L. pneumophila but inhibit intracellular replication of the bacteria and participate in the modulation of the LCV. Uptake and degradation of an icmT mutant strain lacking a functional Icm/Dot transporter was promoted by PI3Ks. We identified Icm/Dot-secreted proteins which specifically bind to phosphatidylinositol(4) phosphate (PI(4)P) in vitro and preferentially localize to LCVs in the absence of functional PI3Ks. PI(4)P was found to be present on LCVs using as a probe either an antibody against PI(4)P or the PH domain of the PI(4)P-binding protein FAPP1 (phosphatidylinositol(4) phosphate adaptor protein 1). Moreover, the presence of PI(4)P on LCVs required a functional Icm/Dot T4SS. Our results indicate that L. pneumophila modulates host cell PI metabolism and exploits the Golgi lipid second messenger PI(4)P to anchor secreted effector proteins to the LCV. PMID- 16710456 TI - Mapping the literature of emergency nursing. AB - PURPOSE: Emergency nursing covers a broad spectrum of health care from trauma surgery support to preventive health care. The purpose of this study is to identify the core literature of emergency nursing and to determine which databases provide the most thorough indexing access to the literature cited in emergency nursing journals. This study is part of the Medical Library Association's Nursing and Allied Health Resources Section's project to map the nursing literature. METHODS: Four key emergency nursing journals were selected and subjected to citation analysis based on Bradford's Law of Scattering. RESULTS: A group of 12 journals made up 33.3% of the 7,119 citations, another 33.3% of the citations appeared in 92 journals, with the remaining 33.3% scattered across 822 journals. Three of the core 12 journals were emergency medicine titles, and 2 were emergency nursing titles from the selected source journals. Government publications constituted 7.5% of the literature cited. CONCLUSIONS: PubMed/MEDLINE provided the best overall indexing coverage for the journals, followed by CINAHL. However, CINAHL provided the most complete coverage for the source journals and the majority of the nursing and emergency medical technology publications and should be consulted by librarians and nurses seeking emergency nursing literature. PMID- 16710457 TI - Mapping the literature of nurse practitioners. AB - OBJECTIVES: This study was designed to identify core journals for the nurse practitioner specialty and to determine the extent of their indexing in bibliographic databases. METHODS: As part of a larger project for mapping the literature of nursing, this study followed a common methodology based on citation analysis. Four journals designated by nurse practitioners as sources for their practice information were selected. All cited references were analyzed to determine format types and publication years. Bradford's Law of Scattering was applied to identify core journals. Nine bibliographic databases were searched to estimate the index coverage of the core titles. RESULTS: The findings indicate that nurse practitioners rely primarily on journals (72.0%) followed by books (20.4%) for their professional knowledge. The majority of the identified core journals belong to non-nursing disciplines. This is reflected in the indexing coverage results: PubMed/MEDLINE more comprehensively indexes the core titles than CINAHL does. CONCLUSION: Nurse practitioners, as primary care providers, consult medical as well as nursing sources for their information. The implications of the citation analysis findings are significant for collection development librarians and indexing services. PMID- 16710458 TI - Mapping the literature of nursing education. AB - OBJECTIVES: As part of a project to map the literature of nursing, sponsored by the Nursing and Allied Health Resources Section of the Medical Library Association, this study identifies core journals cited in nursing education journals and the indexing services that cover the cited journals. METHODS: Three nursing education source journals were subjected to a citation analysis of articles from 1997 to 1999, followed by an analysis of database access to the most frequently cited journal titles. RESULTS: Cited formats included journals (62.4%), books (31.3%), government documents (1.4%), Internet (0.3%), and miscellaneous (4.6%). Cited references were relatively older than other studies, with just 58.6% published in the 1990s. One-third of the citations were found in a core of just 6 journal titles; one-third were dispersed among a middle zone of 53 titles; the remaining third were scattered in a larger zone of 762 titles. Indexing coverage for the core titles was most comprehensive in CINAHL, followed by PubMed/MEDLINE and Social Sciences Citation Index. CONCLUSIONS: Citation patterns in nursing education show more reliance on nursing and education literature than biomedicine. Literature searches need to include CINAHL and PubMed/MEDLINE, as well as education and social sciences databases. Likewise, library collections need to include education and social sciences resources to complement works developed for nurse educators. PMID- 16710459 TI - Mapping the literature of pediatric nursing. AB - BACKGROUND: Pediatric nurses work in an interdisciplinary field and face ever increasing demands on their time and knowledge. Selection tools for librarians serving this group are available, but only one bibliometric analysis has examined citations to aid collection development. METHOD: The "Mapping the Literature of Nursing Project" protocol was used. Three source journals were selected, and a citation analysis of articles from 1998 to 2000 was conducted. RESULTS: The frequency of journal citation was tabulated, and a list of the most frequently cited journals was created. Just over 1% of the cited journals produced 33% of the citations. PubMed/MEDLINE, Science Citation Index, and Social Sciences Citation Index provided the most complete indexing coverage of all types of the journals, while CINAHL providing the most complete coverage of nursing journals. Books were the second-most frequently cited format. CONCLUSIONS: Citation analysis of journal articles from pediatric nursing journals may be helpful in selecting journals for libraries serving pediatric nurses and those who conduct pediatric nursing research. Librarians should consider adding indexes to their collection in addition to PubMed/MEDLINE to access the broad range of journals useful to this specialty. PMID- 16710460 TI - Mapping the literature of rehabilitation nursing. AB - OBJECTIVE: This paper describes a citation analysis of the literature of rehabilitation nursing, conducted as part of the Medical Library Association's Nursing and Allied Health Section's the "Mapping the Literature of Nursing Project." METHODS: One core journal, Rehabilitation Nursing, was selected, being both the official journal of the Association of Rehabilitation Nurses and the only journal devoted exclusively to rehabilitation nursing. Citations were analyzed according to format and date and stratified according to Bradford's Law of Scattering. RESULTS: The nineteen journals that constitute Zone 1 contribute the same number of citations as the eighty-six journals that make up Zone 2. OCLC ArticleFirst, PubMed/MEDLINE, and CINAHL provide the most inclusive coverage of the rehabilitation literature. The source journal, Rehabilitation Nursing, is the most important journal in Zone 1 and thus the most influential rehabilitation nursing journal. Relative degrees of database coverage do not change between Zones 1 and 2. CONCLUSION: The journals in Zones 1 and 2 collectively represent most of the important subspecialties of rehabilitation nursing, such as the physiological, sociopsychological, and community reintegration issues involved in the long-term rehabilitation process. PMID- 16710461 TI - Mapping the literature of transcultural nursing. AB - OVERVIEW: No bibliometric studies of the literature of the field of transcultural nursing have been published. This paper describes a citation analysis as part of the project undertaken by the Nursing and Allied Health Resources Section of the Medical Library Association to map the literature of nursing. OBJECTIVE: The purpose of this study was to identify the core literature and determine which databases provided the most complete access to the transcultural nursing literature. METHODS: Cited references from essential source journals were analyzed for a three-year period. Eight major databases were compared for indexing coverage of the identified core list of journals. RESULTS: This study identifies 138 core journals. Transcultural nursing relies on journal literature from associated health sciences fields in addition to nursing. Books provide an important format. Nearly all cited references were from the previous 18 years. In comparing indexing coverage among 8 major databases, 3 databases rose to the top. CONCLUSIONS: No single database can claim comprehensive indexing coverage for this broad field. It is essential to search multiple databases. Based on this study, PubMed/MEDLINE, Social Sciences Citation Index, and CINAHL provide the best coverage. Collections supporting transcultural nursing require robust access to literature beyond nursing publications. PMID- 16710462 TI - Mapping the general literature of American nursing. AB - OBJECTIVES: As part of a project to map the literature of nursing, sponsored by the Nursing and Allied Health Resources Section of the Medical Library Association, this study identifies core journals cited by general or "popular" US nursing journals and the indexing services that cover the cited journals. METHODS: Three journals were selected for analysis: American Journal of Nursing, Nursing 96-98, and RN. The source journals were subjected to a citation analysis of articles from 1996 to 1998, followed by an analysis of database access to the most frequently cited journal titles. RESULTS: Cited formats included journals (63.7%), books (26.6%), government documents (3.0%), Internet (0.5%), and miscellaneous (6.2%). Cited references were relatively current; most (86.6%) were published in the current decade. One-third of the citations were found in a core of 24 journal titles; one-third were dispersed among a middle zone of 94 titles; and the remaining third were scattered in a larger zone of 694 titles. Indexing coverage for the core titles was most comprehensive in PubMed/MEDLINE, followed by CINAHL and Science Citation Index. CONCLUSIONS: Results support the popular (not scholarly) nature of these titles. While not a good source for original research, they fulfill a key role of disseminating nursing knowledge with their relevantly current citations to a broad variety of sources. PMID- 16710463 TI - Mapping the literature of home health nursing. AB - OBJECTIVES: The purpose of this study was to identify core journals in home health nursing and to determine how well these journals were covered by indexing and abstracting services. The study was part of the project for mapping the nursing literature of the Medical Library Association's Nursing and Allied Health Resource Section. METHODS: A citation analysis of two core journals was done to determine distribution of references by format types and age of citations and dispersion of the literature, according to Bradford's Law of Scattering. The analysis of indexing coverage for Zone 1 and 2 was also provided. RESULTS: The study showed that 64.2% of citations came from journals, versus 22.9% from books and 12.9% from other publications. PubMed/ MEDLINE rated highest in average indexing coverage of Zone 1 and 2 journals, followed by CINAHL. PsycINFO, SocioAbstracts, and EBSCO Health Business FullTEXT showed practically no coverage for the home health nursing literature. CONCLUSION: As expected, journal articles were found to be the primary source for referencing and books, the secondary source. In regard to bibliographic control, no databases provided full coverage of the journals in the field of home health nursing. PubMed/MEDLINE and CINAHL gave better results in combination, because CINAHL tended to cover more nursing journals, while PubMed/MEDLINE did better with medical titles. PMID- 16710464 TI - Mapping the literature of maternal-child/gynecologic nursing. AB - OBJECTIVES: As part of a project to map the literature of nursing, sponsored by the Nursing and Allied Health Resources Section of the Medical Library Association, this study identifies core journals cited in maternal child/gynecologic nursing and the indexing services that access the cited journals. METHODS: Three source journals were selected and subjected to a citation analysis of articles from 1996 to 1998. RESULTS: Journals were the most frequently cited format (74.1%), followed by books (19.7%), miscellaneous (4.2%), and government documents (1.9%). Bradford's Law of Scattering was applied to the results, ranking cited journal references in descending order. One-third of the citations were found in a core of 14 journal titles; one-third were dispersed among a middle zone of 100 titles; and the remaining third were scattered in a larger zone of 1,194 titles. Indexing coverage for the core titles was most comprehensive in PubMed/MEDLINE, followed by Science Citation Index and CINAHL. CONCLUSION: The core of journals cited in this nursing specialty revealed a large number of medical titles, thus, the biomedical databases provide the best access. The interdisciplinary nature of maternal-child/ gynecologic nursing topics dictates that social sciences databases are an important adjunct. The study results will assist librarians in collection development, provide end users with guidelines for selecting databases, and influence database producers to consider extending coverage to identified titles. PMID- 16710465 TI - Mapping the literature of medical-surgical nursing. AB - BACKGROUND: Medical-surgical or adult health nursing is a complex specialty that requires a wide-ranging literature to inform its research and practice. Several excellent qualitative aids exist for collection development for this field, but quantitative studies are few. While one bibliometric study of journals exists, no recent work had been done in this area. METHOD: The Mapping the Literature of Nursing Project protocol was used. Four source journals were selected, and a citation analysis of articles from 1996 to 1998 was conducted. RESULTS: A list of the most frequently cited journals was created, using Bradford's Law of Scattering. The list demonstrates that 1.2% of the cited medical-surgical nursing journals produced just over 33% of the citations. PubMed/MEDLINE, CINAHL, and Science Citation Index provided the most complete indexing coverage of all of the journals, with CINAHL providing the most complete coverage of nursing journals. Books were the second-most cited format. CONCLUSIONS: Citation analysis of journal articles is a useful aid for selecting journals for medical-surgical nursing collections, but it did not prove to be as useful for selecting materials in other formats. Indexes in addition to PubMed/MEDLINE are necessary to provide access to the journal literature serving this specialty. PMID- 16710467 TI - Mapping the literature of nurse-midwifery. AB - OBJECTIVE: This article is part of a project for mapping the literature of nursing. The purpose is to identify the core journals in nurse-midwifery and to determine the extent to which these titles are covered by standard indexing sources. METHODS: Cited references from two source journals were analyzed to discover the most frequently cited publications, including their format, age, and amount of dispersion, as well as the indexes that offer the most complete coverage. RESULTS: A study of the literature of nurse-midwifery reveals that the field is diverse, including such topics as women's health, obstetrics, gynecology, and parent-child relations, in addition to the practice of nurse midwifery itself. Journals were the most heavily cited format, and analysis revealed that ten journals provided one-third of all the references in the study. CONCLUSIONS: A combination of Science Citation Index and Social Sciences Citation Index or PubMed/MEDLINE provided the best overall coverage because of the field's reliance on medical journals. CINAHL had the most complete coverage for the nurse midwifery journals as well as nursing journals in general. PMID- 16710466 TI - Mapping the literature of nephrology nursing. AB - OBJECTIVES: A bibliometric investigation was done to identify characteristics of the literature that nephrology nurses utilize. It is one component of a broader study, "Mapping the Literature of Nursing," by the Medical Library Association's Nursing and Allied Health Resources Section Task Force to Map the Literature of Nursing. METHODS: Following a standard protocol, this project utilized Bradford's Law of Scattering to analyze the literature of nephrology nursing. Citation analysis was done on articles that were published from 1996 to 1998 in a source journal. Cited journal titles were divided into three zones, and coverage in major article databases were scored for Zones 1 and 2. RESULTS: During the three year period, journals were the most frequently cited format type. Eighty-one journals were cited in Zones 1 and 2. As Bradford's Law of Scattering predicted, a small number of the cited journals accounted for the most use. Coverage is most comprehensive for cited journals in Science Citation Index, PubMed/ MEDLINE, and EMBASE. When looking just at cited nursing journals, CINAHL and PubMed/MEDLINE provide the best indexing coverage. CONCLUSION: This study offers understanding of and insights into the types of information that nephrology nurses use for research. It is a valuable tool for anyone involved with providing nephrology nursing literature. PMID- 16710468 TI - Mapping the literature of nursing administration. AB - OBJECTIVES: As part of Phase I of a project to map the literature of nursing, sponsored by the Nursing and Allied Health Resources Section of the Medical Library Association, this study identifies the core literature cited in nursing administration and the indexing services that provide access to the core journals. The results of this study will assist librarians and end users searching for information related to this nursing discipline, as well as database producers who might consider adding specific titles to their indexing services. METHODS: Using the common methodology described in the overview article, five source journals for nursing administration were identified and selected for citation analysis over a three-year period, 1996 to 1998, to identify the most frequently cited titles according to Bradford's Law of Scattering. From this core of most productive journal titles, the bibliographic databases that provide the best access to these titles were identified. RESULTS: Results reveal that nursing administration literature relies most heavily on journal articles and on those titles identified as core nursing administrative titles. When the indexing coverage of nine services is compared, PubMed/MEDLINE and CINAHL provide the most comprehensive coverage of this nursing discipline. CONCLUSIONS: No one indexing service adequately covers this nursing discipline. Researchers needing comprehensive coverage in this area must search more than one database to effectively research their projects. While PubMed/MEDLINE and CINAHL provide more coverage for this discipline than the other indexing services, none is sufficiently broad in scope to provide indexing of nursing, health care management, and medical literature in a single file. Nurse administrators using the literature to research current work issues need to review not only the nursing titles covered by CINAHL but should also include the major weekly medical titles, core titles in health care administration, and general business sources if they wish to adequately cover the many aspects of nursing administration. PMID- 16710469 TI - Mapping the literature of nursing informatics. AB - OBJECTIVE: This study was part of the Medical Library Association's Nursing and Allied Health Resources Section's project to map the nursing literature. It identified core journals in nursing informatics and the journals referenced in them and analyzed coverage of those journals in selected indexes. METHOD: Five core journals were chosen and analyzed for 1996, 1997, and 1998. The references in the core journal articles were examined for type and number of formats cited during the selected time period. Bradford's Law of Scattering divided the journals into frequency zones. RESULTS: The time interval, 1990 to 1998, produced 71% of the references. Internet references could not be tracked by date before 1990. Twelve journals were the most productive, 119 journals were somewhat productive, and 897 journals were the least productive. Journal of the American Medical Informatics Association was the most prolific core journal. The 1998 journal references were compared in CINAHL, PubMed/MEDLINE, Science Citation Index, and OCLC Article First. PubMed/MEDLINE had the highest indexing score. PMID- 16710470 TI - Mapping the literature of case management nursing. AB - OBJECTIVES: Nursing case management provides a continuum of health care services for defined groups of patients. Its literature is multidisciplinary, emphasizing clinical specialties, case management methodology, and the health care system. This study is part of a project to map the literature of nursing, sponsored by the Nursing and Allied Health Resources Section of the Medical Library Association. The study identifies core journals cited in case management literature and indexing services that access those journals. METHODS: Three source journals were identified based on established criteria, and cited references from each article published from 1997 to 1999 were analyzed. RESULTS: Nearly two-thirds of the cited references were from journals; others were from books, monographs, reports, government documents, and the Internet. Cited journal references were ranked in descending order, and Bradford's Law of Scattering was applied. The many journals constituting the top two zones reflect the diversity of this field. Zone 1 included journals from nursing administration, case management, general medicine, medical specialties, and social work. Two databases, PubMed/MEDLINE and OCLC ArticleFirst, provided the best indexing coverage. CONCLUSION: Collections that support case management require a relatively small group of core journals. Students and health care professionals will need to search across disciplines to identify appropriate literature. PMID- 16710472 TI - Ectopic pancreas formation in Hes1 -knockout mice reveals plasticity of endodermal progenitors of the gut, bile duct, and pancreas. AB - Ectopic pancreas is a developmental anomaly occasionally found in humans. Hes1, a main effector of Notch signaling, regulates the fate and differentiation of many cell types during development. To gain insights into the role of the Notch pathway in pancreatic fate determination, we combined the use of Hes1-knockout mice and lineage tracing employing the Cre/loxP system to specifically mark pancreatic precursor cells and their progeny in Ptf1a-cre and Rosa26 reporter mice. We show that inactivation of Hes1 induces misexpression of Ptf1a in discrete regions of the primitive stomach and duodenum and throughout the common bile duct. All ectopic Ptf1a-expressing cells were reprogrammed, or transcommitted, to multipotent pancreatic progenitor status and subsequently differentiated into mature pancreatic exocrine, endocrine, and duct cells. This process recapitulated normal pancreatogenesis in terms of morphological and genetic features. Furthermore, analysis of Hes1/Ptf1a double mutants revealed that ectopic Ptf1a-cre lineage-labeled cells adopted the fate of region appropriate gut epithelium or endocrine cells similarly to Ptf1a-inactivated cells in the native pancreatic buds. Our data demonstrate that the Hes1-mediated Notch pathway is required for region-appropriate specification of pancreas in the developing foregut endoderm through regulation of Ptf1a expression, providing novel insight into the pathogenesis of ectopic pancreas development in a mouse model. PMID- 16710471 TI - SOCS-3 negatively regulates innate and adaptive immune mechanisms in acute IL-1 dependent inflammatory arthritis. AB - RA is an autoimmune disease characterized by sustained imbalance between pro- and antiinflammatory immune mechanisms. The SOCS proteins are negative regulators of cytokine signaling, but to date there has been little information on their function in disease. The generation of Socs3(-/Delta vav) mice, which lack SOCS-3 in the hematopoietic and endothelial cell compartment, allowed us to explore the role of endogenous SOCS-3 during acute inflammatory arthritis. Joint inflammation in Socs3(-/Delta vav) mice was particularly severe and was characterized by increased numbers of neutrophils in the inflamed synovium, bone marrow, peripheral blood, and spleen. These features were most likely due to increased production of and enhanced responsiveness to G-CSF and IL-6 during arthritis in these mice. Local osteoclast generation and bone destruction were also dramatically increased in the absence of SOCS-3, as was macrophage activation. Finally, SOCS-3 was found to negatively regulate CD4+ T lymphocyte activation, including production of the pleiotropic cytokine IL-17. The absence of SOCS-3 therefore had dramatic effects in this disease model, with a broader impact on cellular responses than SOCS-1 deficiency. These findings provide direct in vivo evidence that endogenous SOCS-3 is a critical negative regulator of multiple cell types orchestrating inflammatory joint disease. PMID- 16710473 TI - Epimorphin(-/-) mice have increased intestinal growth, decreased susceptibility to dextran sodium sulfate colitis, and impaired spermatogenesis. AB - Dynamic and reciprocal epithelial-mesenchymal interactions are critical for the normal morphogenesis and maintenance of epithelia. Epimorphin has been identified as a unique molecule expressed by mesenchymal cells and myofibroblasts and has putative morphogenetic effects in multiple epithelial tissues, including intestine, skin, mammary gland, lung, gallbladder, and liver. To define the in vivo role of epimorphin, we created epimorphin-null mice by targeted inactivation of the epimorphin gene. Male epimorphin-/- mice are sterile due to abnormal testicular development and impaired spermatogenesis. Intestinal growth is increased in epimorphin-/- mice due to augmented crypt cell proliferation and crypt fission during the neonatal (suckling) period, mediated at least in part by changes in bone morphogenetic protein (Bmp) and Wnt/beta-catenin signaling pathways. Colonic mucosal injury and colitis induced by dextran sodium sulfate (DSS) are ameliorated in epimorphin-/- mice, probably due to the increased proliferative capacity of the epimorphin-/- colon. These in vivo findings support the notion that epimorphin is a key stromal regulator of epithelial cell proliferation and growth in the intestine. In addition, our studies demonstrate a novel and critical role for epimorphin in regulating testicular development and growth as well as spermatogenesis. PMID- 16710474 TI - Longitudinal noninvasive PET-based beta cell mass estimates in a spontaneous diabetes rat model. AB - Diabetes results from an absolute or relative reduction in pancreatic beta cell mass (BCM) leading to insufficient insulin secretion and hyperglycemia. Measurement of insulin secretory capacity is currently used as a surrogate measure of BCM. However, serum insulin concentrations provide an imprecise index of BCM, and no reliable noninvasive measure of BCM is currently available. Type 2 vesicular monoamine transporters (VMAT2) are expressed in human islet beta cells, as well as in tissues of the CNS. [11C]Dihydrotetrabenazine ([11C]DTBZ) binds specifically to VMAT2 and is a radioligand currently used in clinical imaging of the brain. Here we report the use of [11C]DTBZ to estimate BCM in a rodent model of spontaneous type 1 diabetes (the BB-DP rat). In longitudinal PET studies of the BB-DP rat, we found a significant decline in pancreatic uptake of [11C]DTBZ that anticipated the loss of glycemic control. Based on comparison of standardized uptake values (SUVs) of [11C]DTBZ and blood glucose concentrations, loss of more than 65% of the original SUV correlated significantly with the development of persistent hyperglycemia. These studies suggest that PET-based quantitation of VMAT2 receptors provides a noninvasive measurement of BCM that could be used to study the pathogenesis of diabetes and to monitor therapeutic interventions. PMID- 16710475 TI - Neutrophils and their Fc gamma receptors are essential in a mouse model of transfusion-related acute lung injury. AB - Transfusion-related acute lung injury (TRALI) is the most common cause of transfusion-related mortality. To explore the pathogenesis of TRALI, we developed an in vivo mouse model based on the passive transfusion of an MHC class I (MHC I) mAb (H2Kd) to mice with the cognate antigen. Transfusion of the MHC I mAb to BALB/c mice produced acute lung injury with increased excess lung water, increased lung vascular and lung epithelial permeability to protein, and decreased alveolar fluid clearance. There was 50% mortality at a 2-hour time point after Ab administration. Pulmonary histology and immunohistochemistry revealed prominent neutrophil sequestration in the lung microvasculature that occurred concomitantly with acute peripheral blood neutropenia, all within 2 hours of administration of the mAb. Depletion of neutrophils by injection of anti granulocyte mAb Gr-1 protected mice from lung injury following MHC I mAb challenge. FcRgamma-/- mice were resistant to MHC I mAb-induced lung injury, while adoptive transfer of wild-type neutrophils into the FcRgamma-/- animals restored lung injury following MHC I mAb challenge. In conclusion, in a clinically relevant in vivo mouse model of TRALI using an MHC I mAb, the mechanism of lung injury was dependent on neutrophils and their Fc gamma receptors. PMID- 16710476 TI - Met-regulated expression signature defines a subset of human hepatocellular carcinomas with poor prognosis and aggressive phenotype. AB - Identification of specific gene expression signatures characteristic of oncogenic pathways is an important step toward molecular classification of human malignancies. Aberrant activation of the Met signaling pathway is frequently associated with tumor progression and metastasis. In this study, we defined the Met-dependent gene expression signature using global gene expression profiling of WT and Met-deficient primary mouse hepatocytes. Newly identified transcriptional targets of the Met pathway included genes involved in the regulation of oxidative stress responses as well as cell motility, cytoskeletal organization, and angiogenesis. To assess the importance of a Met-regulated gene expression signature, a comparative functional genomic approach was applied to 242 human hepatocellular carcinomas (HCCs) and 7 metastatic liver lesions. Cluster analysis revealed that a subset of human HCCs and all liver metastases shared the Met induced expression signature. Furthermore, the presence of the Met signature showed significant correlation with increased vascular invasion rate and microvessel density as well as with decreased mean survival time of HCC patients. We conclude that the genetically defined gene expression signatures in combination with comparative functional genomics constitute an attractive paradigm for defining both the function of oncogenic pathways and the clinically relevant subgroups of human cancers. PMID- 16710477 TI - Ligation of protease-activated receptor 1 enhances alpha(v)beta6 integrin dependent TGF-beta activation and promotes acute lung injury. AB - Activation of latent TGF-beta by the alpha(v)beta6 integrin is a critical step in the development of acute lung injury. However, the mechanism by which alpha(v)beta6-mediated TGF-beta activation is regulated has not been identified. We show that thrombin, and other agonists of protease-activated receptor 1 (PAR1), activate TGF-beta in an alpha(v)beta6 integrin-specific manner. This effect is PAR1 specific and is mediated by RhoA and Rho kinase. Intratracheal instillation of the PAR1-specific peptide TFLLRN increases lung edema during high tidal-volume ventilation, and this effect is completely inhibited by a blocking antibody against the alpha(v)beta6 integrin. Instillation of TFLLRN during high tidal-volume ventilation is associated with increased pulmonary TGF-beta activation; however, this is not observed in Itgb6-/- mice. Furthermore, Itgb6-/- mice are also protected from ventilator-induced lung edema. We also demonstrate that pulmonary edema and TGF-beta activity are similarly reduced in Par1-/- mice following bleomycin-induced lung injury. These results suggest that PAR1-mediated enhancement of alpha(v)beta6-dependent TGF-beta activation could be one mechanism by which activation of the coagulation cascade contributes to the development of acute lung injury, and they identify PAR1 and the alpha(v)beta6 integrin as potential therapeutic targets in this condition. PMID- 16710478 TI - Immune sensing of Candida albicans requires cooperative recognition of mannans and glucans by lectin and Toll-like receptors. AB - The fungal pathogen Candida albicans has a multilayered cell wall composed of an outer layer of proteins glycosylated with N- or O-linked mannosyl residues and an inner skeletal layer of beta-glucans and chitin. We demonstrate that cytokine production by human mononuclear cells or murine macrophages was markedly reduced when stimulated by C. albicans mutants defective in mannosylation. Recognition of mannosyl residues was mediated by mannose receptor binding to N-linked mannosyl residues and by TLR4 binding to O-linked mannosyl residues. Residual cytokine production was mediated by recognition of beta-glucan by the dectin-1/TLR2 receptor complex. C. albicans mutants with a cell wall defective in mannosyl residues were less virulent in experimental disseminated candidiasis and elicited reduced cytokine production in vivo. We concluded that recognition of C. albicans by monocytes/macrophages is mediated by 3 recognition systems of differing importance, each of which senses specific layers of the C. albicans cell wall. PMID- 16710481 TI - High field ENDOR as a characterization tool for functional sites in microporous materials. AB - The determination of the details of the spatial and electronic structure of functional sites (centers) in any system, be it in materials chemistry or in biology, is the first step towards understanding their function. When such sites happen to be paramagnetic in any point of their activity cycle, the tool box offered by a variety of high resolution electron paramagnetic resonance (EPR) spectroscopic techniques becomes very attractive for their characterization. This tool box has been considerably expanded by the developments in high field (HF) EPR in general, and HF electron nuclear double resonance (ENDOR), in particular. These have led to numerous new applications in the fields of biology, physics, chemistry and materials sciences. This overview focuses specifically on recent applications of pulsed HF ENDOR spectroscopy to microporous materials, such as zeotype materials, presenting the new opportunities it offers. First, a brief description of the theoretical basis required for the analysis of the HF ENDOR spectrum is given, followed by a description of the pulsed techniques used to record spectra and assign the signals, along with a brief presentation of the required instrumentation. Next, specific applications are given, including transition metal ions and complexes exchanged into zeolite cages, transition metal substitution into frameworks of zeolites, aluminophosphate molecular sieves, and silicious mesoporous materials, the interaction of NO with Lewis sites in zeolite cages and trapped S. We end with a discussion of the advantages and the shortcomings of the method and conclude with a future outlook. PMID- 16710479 TI - Reprogramming of antiviral T cells prevents inactivation and restores T cell activity during persistent viral infection. AB - Failure to clear persistent viral infections results from the early loss of T cell activity. A pertinent question is whether the immune response is programmed to fail or if nonresponsive T cells can specifically be fixed to eliminate infection. Although evidence indicates that T cell expansion is permanently programmed during the initial priming events, the mechanisms that determine the acquisition of T cell function are less clear. Herein we show that in contrast to expansion, the functional programming of T cell effector and memory responses in vivo in mice is not hardwired during priming but is alterable and responsive to continuous instruction from the antigenic environment. As a direct consequence, dysfunctional T cells can be functionally reactivated during persistent infection even after an initial program of inactivation has been instituted. We also show that early therapeutic reductions in viral replication facilitate the preservation of antiviral CD4+ T cell activity, enabling the long-term control of viral replication. Thus, dysfunctional antiviral T cells can regain activity, providing a basis for future therapeutic strategies to treat persistent viral infections. PMID- 16710482 TI - Proton tunneling estimates for malonaldehyde vibrations from supersonic jet and matrix quenching experiments. AB - FTIR tunneling splittings for a range of fundamental excitations of malonaldehyde are determined from adiabatic cooling and symmetry breaking experiments in three different molecular states (isolated, coated by Ar layers and embedded in bulk Ar matrices), showing that there is room for improvement in available theoretical models. PMID- 16710483 TI - Comparison of frozen-density embedding and discrete reaction field solvent models for molecular properties. AB - We investigate the performance of two discrete solvent models in connection with density functional theory (DFT) for the calculation of molecular properties. In our comparison we include the discrete reaction field (DRF) model, a combined quantum mechanics and molecular mechanics (QM/MM) model using a polarizable force field, and the frozen-density embedding (FDE) scheme. We employ these solvent models for ground state properties (dipole and quadrupole moments) and response properties (electronic excitation energies and frequency-dependent polarizabilities) of a water molecule in the liquid phase. It is found that both solvent models agree for ground state properties, while there are significant differences in the description of response properties. The origin of these differences is analyzed in detail and it is found that they are mainly caused by a different description of the ground state molecular orbitals of the solute. In addition, for the calculation of the polarizabilities, the inclusion of the response of the solvent to the polarization of the solute becomes important. This effect is included in the DRF model, but is missing in the FDE scheme. A way of including it in FDE calculations of the polarizabilities using finite field calculations is demonstrated. PMID- 16710480 TI - Mast cells can promote the development of multiple features of chronic asthma in mice. AB - Bronchial asthma, the most prevalent cause of significant respiratory morbidity in the developed world, typically is a chronic disorder associated with long-term changes in the airways. We developed a mouse model of chronic asthma that results in markedly increased numbers of airway mast cells, enhanced airway responses to methacholine or antigen, chronic inflammation including infiltration with eosinophils and lymphocytes, airway epithelial goblet cell hyperplasia, enhanced expression of the mucin genes Muc5ac and Muc5b, and increased levels of lung collagen. Using mast cell-deficient (Kit(W-sh/W-sh) and/or Kit(W/W-v)) mice engrafted with FcRgamma+/+ or FcRgamma-/- mast cells, we found that mast cells were required for the full development of each of these features of the model. However, some features also were expressed, although usually at less than wild type levels, in mice whose mast cells lacked FcRgamma and therefore could not be activated by either antigen- and IgE-dependent aggregation of Fc epsilonRI or the binding of antigen-IgG1 immune complexes to Fc gammaRIII. These findings demonstrate that mast cells can contribute to the development of multiple features of chronic asthma in mice and identify both Fc Rgamma-dependent and Fc Rgamma-independent pathways of mast cell activation as important for the expression of key features of this asthma model. PMID- 16710484 TI - The absorption spectra of anisole-h8, anisole-d3 and anisole-d8. The assignment of fundamental vibrations in the S0 and the S1 states. AB - The REMPI spectra of anisole-h8, anisole-d3 and anisole-d8 have been measured. The assignment of the fundamental vibrations of anisole in the S(1) state is supported by quantum chemical model calculations, the isotopic shifts, the comparison with the frequencies of corresponding vibrations in other monosubstituted benzenes, especially phenol, and the overtones, combinations and progressions observed in the spectrum of anisole-h8. The frequencies of the 42 fundamental vibrations of anisole in the S(1) state are evaluated and compared with the frequencies of the corresponding vibrations in the electronic ground state. Some assignments given earlier in the literature have been revised. PMID- 16710485 TI - Acid sites and oxidation center in molybdena supported on tin oxide as studied by solid-state NMR spectroscopy and theoretical calculation. AB - Solid-state NMR spectroscopy and density functional theory (DFT) calculations were employed to study the structure and properties, especially the solid acidity, of molybdenum oxide supported on tin oxide. As demonstrated by solid state NMR experiments, Mo species are mainly dispersed on the surface of SnO(2) support rather than significantly dissolved into the SnO(2) structure and Bronsted as well as Lewis acid sites are present on the MoO(3)/SnO(2) catalyst. Acid strength of the supported metal oxide is stronger than those of zeolites, e.g., HY and HZSM-5, though the concentration of acid sites is relatively lower. The DFT calculated (13)C chemical shift for acetone adsorbed on MoO(3)/SnO(2) is in good agreement with the experimental value, which confirms our proposed structure of -Mo-(OH)-Sn- for the Bronsted acid site. Reducibility of the supported metal oxide is also demonstrated by solid-state NMR experiments and an active oxidation center of this catalyst is proposed as well. PMID- 16710486 TI - Hydrogen adsorption kinetics on Pd/Ce0.8Zr0.2O2. AB - Hydrogen adsorption on Pd/Ce(0.8)Zr(0.2)O(2) was studied by temperature programmed reduction, volumetric measurements and IR spectroscopy. Hydrogen uptake and reduction rate at 353 K are strongly dependent on the hydrogen pressure. At relatively high hydrogen partial pressure, reduction involves PdO, the surface and a significant fraction of the bulk of the ceria based oxide. Formation of oxygen vacancies even at low temperature (<373 K) is observed. The hydrogen adsorption process is mainly irreversible, as is shown by an increase in the (2)F(5/2)-->(2)F(7/2) electronic transition of Ce(3+) with hydrogen pressure and surface dehydroxylation. This "severe" reduction has a negative effect on the subsequent hydrogen adsorption capability. The decrease of hydrogen uptake capacity and rate during adsorption can be associated with the partial loss of superficial OH and the presence of Ce(3+), which deactivates Pd electronically. PMID- 16710487 TI - C-O bond scission of methoxide on Pd nanoparticles: a density functional study. AB - C-O bond scission of methoxide species adsorbed at the surface of Pd nanoparticle was studied by DF calculations for the example of cuboctahedral Pd(79). To investigate different locations of adsorbed intermediates as well as the transition state of C-O bond scission, a substrate model was used, which allows one to consider adsorbates without any local geometry restrictions. In contrast to reaction sites on the flat Pd(111) surface and on extended facets, scission of the C-O bond of methoxide at cluster edges is exothermic by approximately 40 kJ mol(-1) and the decomposition product CH(3) is found to be stabilized there. However, the high calculated activation barrier, approximately 140 mol(-1), implies only a very slow reaction compared to dehydrogenation of CH(3)O. PMID- 16710488 TI - Origin of the bathochromically shifted optical spectra of meso-tetrathien-2'- and 3'-ylporphyrins as compared to meso-tetraphenylporphyrin. AB - The UV-Vis and fluorescence spectra of free base and diprotonated meso-tetrathien 2'-ylporphyrins are, when compared to the spectra of meso-tetra-phenyl- or even thien-3'-ylporphyrins, characterized by surprisingly large red-shifts. A comparison of the optical spectra and the computed rotational barriers for these meso-aryl-substituted porphyrins and a detailed conformational analysis of the single crystal X-ray structure of a diprotonated meso-tetrathien-2'-ylporphyrin suggest that the origin of the altered electronic properties of meso-tetrathien 2'-ylporphyrins are mainly due to the contribution of conformations in which the thienyl groups adopt idealized co-planar arrangements with the porphyrin ring. These conformations allow an efficient extension of the porphyrinic pi-system through conjugation. We synthesized a meso-tetrathien-2'-ylporphyrin with methyl groups in the o-position, thus preventing the formation of conformers with co planar thienyl groups and a corresponding thien-2'-ylporphyrin with methyl substituents in a distal position that possesses the same steric requirements for thienyl group rotation as the parent compound, to conclusively deduce the influence of the conformers on the electronic structure. A MNDO-PSDCI computation of their optical spectra further supports our key hypothesis. DFT computations of the total energies of the hypothetical diprotonated thien-2'-ylporphyrin conformer with perpendicular thienyl groups and the conformer containing near-co planar thienyl groups quantify the resonance stabilization energy. Our results support and complement recent photophysical and theoretical studies by Gupta and Ravikanth and Friedlein et al. on thien-2'-yl-substituted core-modified porphyrins and [meso-tetra(5'-bromothien-2'-yl)porphyrinato]Zn(ii), respectively. PMID- 16710489 TI - Dealing with a local heating effect when measuring catalytic solids in a reactor with Raman spectroscopy. AB - In continuation of our previous work on the applicability of the G(R(infinity)) correction factor for the quantification of Raman spectra of coke during propane dehydrogenation experiments (Phys. Chem. Chem. Phys., 2005, 7, 211), research has been carried out on the potential of this correction factor for the quantification of supported metal oxides during reduction experiments. For this purpose, supported chromium oxide catalysts have been studied by combined in situ Raman and UV-Vis spectroscopy during temperature programmed reduction experiments with hydrogen as reducing agent. The goal was to quantify on-line the amount of Cr(6+) in a reactor based on the measured in situ Raman spectra. During these experiments, a significant temperature effect was observed, which has been investigated in more detail with a thermal imaging technique. The results revealed a temperature 'on the spot' that can exceed 100 degrees C. It implies that Raman spectroscopy can have a considerable effect on the local reaction conditions and explains observed inconsistencies between the in situ UV-Vis and Raman data. In order to minimize this heating effect, reduction of the laser power, mathematical matching of the spectroscopic data, a different cell design and a change in reaction conditions has been evaluated. It is demonstrated that increasing the reactor temperature is the most feasible method to solve the heating problem. Next, it allows the application of in situ Raman spectroscopy in a reliable quantitative way without the need of an internal standard. PMID- 16710491 TI - Commercial renal transplantation-- body parts for sale. PMID- 16710490 TI - Influence of Sn4+ on the structural and electronic properties of Ti1-xSnxO2 nanoparticles used as photocatalysts. AB - Ti(1-x)Sn(x)O(2) nanocrystalline materials employed for photocatalysis have been characterised by means of X-ray diffraction, Raman, X-ray absorption (XANES and EXAFS) and UV-Vis spectroscopy and high resolution transmission electron microscopy. Single-phase samples with anatase or rutile type structures and similar tin contents permitted a separate study of the effect of Sn(4+) ions on these crystalline forms, whereas materials composed of anatase and rutile mixtures were used to investigate the distribution of the dopant cations when both phases coexist. The results obtained from the single-phase doped TiO(2) samples indicate that the presence of tin causes a different effect when doping anatase or rutile in both their structural and electronic properties. While a random substitution of Sn(4+) for Ti(4+) seems plausible for the rutile phase, some kind of gradient in Sn(4+) concentration is possible in anatase. On the other hand, when anatase and rutile coexist, effects of doping are visible in both phases. Regarding chemical composition, a homogeneous distribution of tin was found in both calcined and hydrothermal multiphase samples. Photocatalytic experiments show that both tin-doping and coexistence of different phases have a beneficial effect on the activity of the catalysts. PMID- 16710492 TI - Rising trends of STIs and HIV infection in Singapore-- a review of epidemiology over the last 10 years (1994 to 2003). AB - INTRODUCTION: This is a review of the epidemiology and trends of sexually transmitted infections (STIs) in Singapore from 1994 to 2003. MATERIALS AND METHODS: Data collated for both notifiable and non-notifiable STIs from 1994 to 2003 were analysed. This data consisted of STI notifications from medical practitioners in Singapore as well as from the Department of STI Control clinic itself. RESULTS: There was a decline in the overall STI incidence in Singapore in the first half of the last decade from 215 cases per 100,000 population (7,200 cases) in 1994 to 162 cases per 100,000 population (6,318 cases) in 1999, followed by an increasing trend in the number of acute STIs (both bacterial and viral) over the past 5 years to 195 cases per 100,000 population (8,175 cases) in 2003 (P <0.001). The incidence of HIV has risen sharply over the last decade whilst that of other viral STIs has not decreased. Singaporeans are becoming sexually active at a younger age, with casual partners constituting the main primary contacts. CONCLUSIONS: Although there has been a significant decline in the overall incidence of STIs in Singapore over the last decade, a rise in acute STIs over the last 5 years has resulted in the need to identify the causal factors, and to intensify existing as well as develop new STI/HIV prevention programmes for the general population and certain core groups. PMID- 16710493 TI - Modelling the utility of body temperature readings from primary care consults for SARS surveillance in an army medical centre. AB - INTRODUCTION: There is interest in surveillance systems for outbreak detection at stages where clinical presentation would still be undifferentiated. Such systems focus on detecting clusters of syndromes in excess of baseline levels, which may indicate an outbreak. We model the detection limits of a potential system based on primary care consults for the detection of an outbreak of severe acute respiratory syndrome (SARS). MATERIALS AND METHODS: Data from an averaged-sized medical centre were extracted from the Patient Care Enhancement System (PACES) [the electronic medical records system serving the Singapore Armed Forces (SAF)]. Thresholds were set to 3 or more cases presenting with particular syndromes and a temperature reading of >or=38oC (T >or=38). Monte Carlo simulation was used to insert simulated SARS outbreaks of various sizes onto the background incidence of febrile cases, accounting for distribution of SARS incubation period, delay from onset to first consult, and likelihood of presenting with T >or=38 to the SAF medical centre. RESULTS: Valid temperature data was available for 2,012 out of 2,305 eligible syndromic consults (87.2%). T >or=38 was observed in 166 consults (8.3%). Simulated outbreaks would peak 7 days after exposure, but, on average, signals at their peak would consist of 10.9% of entire outbreak size. Under baseline assumptions, the system has a higher than 90% chance of detecting an outbreak only with 20 or more cases. CONCLUSIONS: Surveillance based on clusters of cases with T >or=38 helps reduce background noise in primary care data, but the major limitation of such systems is that they are still only able to confidently detect large outbreaks. PMID- 16710494 TI - Accuracy of fine needle aspiration cytology and frozen section histopathology for lesions of the major salivary glands. AB - INTRODUCTION: Identifying malignancy either preoperatively or intraoperatively can have a significant impact on the management of salivary gland tumours. We review our experience with fine needle aspiration cytology (FNAC) and frozen section (FS) for salivary gland lesions. We analyse the accuracy of both modalities and their influence on management. MATERIALS AND METHODS: Retrospective review of 114 patients who underwent salivary gland surgery, 91 with intraoperative FS and 68 with preoperative FNAC. Both sets of results were compared against each other and the final histopathological diagnosis. RESULTS: The accuracy of FS was 92.3%, with a sensitivity and specificity of 62.5% and 100%. Histologic concordance was 92.4% for benign lesions, and 100% for malignant tumours. The accuracy of FNAC was 89.7%, with a sensitivity and specificity of 100%. The non-diagnostic rate was 10.3%. Histologic concordance for FNAC was inferior to that for FS, with only 64.2% of benign lesions and 50% of malignant tumours correctly identified. FNAC did not alter the management of benign disease even when a correct diagnosis was obtained. CONCLUSION: Our results suggest that FNAC and FS are complementary in usefulness for malignant tumours. However, FNAC does not influence the management of benign lesions and routine FNAC for every patient may not be cost-effective. PMID- 16710495 TI - Clinical assessment of absence of the palmaris longus and its association with other anatomical anomalies-- a Chinese population study. AB - INTRODUCTION: Ethnic variations in the prevalence of absence of the palmaris longus (PL) are well known. Many techniques for clinically determining the presence of the PL have been described. Studies have also attempted to correlate its absence with other anatomical anomalies. However, most studies have been done in Caucasian populations. MATERIALS AND METHODS: The presence of the PL was clinically determined in 329 normal Chinese men and women using the standard technique. In subjects with an absent PL, 4 other tests were performed to confirm absence and an Allen's test was done to assess the palmar arches. All subjects were examined for the presence of the flexor digitorum superficialis (FDS) to the little finger. RESULTS: The overall prevalence of absence of the PL was 4.6%. All techniques were equally effective in determining the absence of the PL. There was no significant difference in its absence with regard to the body side or sex. Absolute deficiency of FDS to the little finger was seen in 6.4%. No correlation could be detected between the absence of the PL and FDS of the little finger. CONCLUSIONS: The prevalence of absence of the PL and absence of FDS to the little finger in a Chinese population is much lower compared to previous reports in the literature. There is no association between absence of the PL and absence of the FDS to the little finger. Although all techniques of examining for the absence of the PL are equally effective, the method suggested by Mishra seems the best as it was easily understood by subjects and can be used even when thumb abduction is not possible. PMID- 16710496 TI - Are sensory and cognitive declines associated in older persons seeking aged care services? Findings from a pilot study. AB - INTRODUCTION: To assess the association between sensory and cognitive function, and the potential influence of visual function on cognitive function assessment, in a sample of clients accessing aged care services. MATERIALS AND METHODS: We recruited 260 non-institutionalised, frail, older individuals who sought aged care services. Visual acuity was assessed using a LogMAR chart and hearing function was measured using a portable pure-tone air conduction audiometer. Visual impairment was defined as visual acuity (VA) <6/12 (<39 letters read correctly in the better eye), moderate-to-severe hearing impairment as hearing thresholds >40 decibels (better ear) and cognitive impairment as Mini-Mental State Examination (MMSE) score <24. RESULTS: Vision and hearing assessments were randomly performed in 168 and 164 aged care clients, respectively. Visual acuity correlated weakly with MMSE scores, either including (r = 0.27, P <0.001) or excluding (r = 0.21, P = 0.006) vision-related MMSE items. After partialling out the effect of age, the association remained (r = 0.23, P = 0.013 including, or r = 0.18, P = 0.044 excluding vision-related items). No correlation was found between MMSE scores and hearing thresholds (r = -0.07, P = 0.375). After adjusting for age, sex and stroke, mean MMSE scores were lower in persons with visual impairment than those with normal vision (25.2 + 0.5 versus 26.8 + 0.4 including, or 18.2 + 0.5 versus 19.2 + 0.3 excluding vision-related items), but were similar between subjects with none or mild and those with moderate-to-severe hearing loss (26.3 + 0.4 versus 26.0 + 0.4). CONCLUSIONS: In this study sample, visual and cognitive functions were modestly associated, after excluding the influence of visual impairment on the MMSE assessment and adjusting for age. Hearing thresholds were not found to be associated with cognitive function. PMID- 16710497 TI - Retrobulbar alcohol injection for orbital pain relief under difficult circumstances: a case report. AB - INTRODUCTION: A case is presented of a patient with severe and chronic pain in the orbital region, which was relieved by retrobulbar injection of absolute alcohol. The management of chronic pain in the orbital region has received little attention and the literature is reviewed. CLINICAL PICTURE: A 52-year-old man with adenoid cystic carcinoma of the maxillary sinus was suffering from severe pain, especially in the left orbital region. There was also pain from ocular exposure and compression caused by the tumour. Magnetic resonance imaging (MRI) revealed spread of tumour to both orbits, particularly on the left. Multiple debulking surgery and various treatment modalities offered no relief from his pain. TREATMENT: A single retrobulbar injection of 2 mL of absolute alcohol, was placed into the putative orbital apex. OUTCOME: As a consequence of the injection, he had complete resolution of his pain in the 6 months prior to his death. DISCUSSION: Retrobulbar injection of alcohol offers effective pain relief in certain specific conditions characterised by chronic orbital pain when other treatments do not help. CONCLUSION: We have demonstrated that the retrobulbar technique still has a place in our armamentarium. PMID- 16710498 TI - Concurrent intermediate uveitis and an enhancing intracranial lesion as the initial manifestation of sarcoidosis. AB - INTRODUCTION: Posterior segment involvement has been described to be associated with central nervous system involvement in sarcoidosis as a result of direct sarcoid tissue infiltration or mass effect of a cerebral lesion. However, isolated intermediate uveitis occurring concurrently with central nervous system involvement prior to extensive systemic disease is rare. CLINICAL PICTURE: We describe a patient with neuro-ophthalmic manifestations of intermediate uveitis and an enhancing basal ganglia lesion at initial presentation, in the absence of extensive systemic disease. TREATMENT: He was treated with high-dose systemic steroids which was progressively tailed down over 6 months. OUTCOME: There was prompt resolution of vitritis with good preservation of visual acuity. CONCLUSION: The difficulties of the initial diagnosis of sarcoidosis and the indications for initiation of steroid therapy are illustrated. We use this case to emphasise the need for a high clinical suspicion of sarcoidosis in the presence of similar unusual and seemingly unrelated combinations of neurological manifestations so as to facilitate the prompt institution of appropriate treatment when indicated. PMID- 16710499 TI - Necrotising fasciitis and traditional medical therapy-- a dangerous liaison. AB - INTRODUCTION: Necrotising fasciitis is a disease associated with high morbidity and mortality, and multi-focal necrotising fasciitis is uncommon. We present 2 cases of concurrent necrotising fasciitis of contralateral upper and lower limbs. CLINICAL PICTURE: Both presented with pain, swelling, bruising or necrosis of the affected extremities. Traditional medical therapy was sought prior to their presentation. TREATMENT: After initial debridement, one patient subsequently underwent amputation of the contralateral forearm and leg. The other underwent a forearm amputation, but refused a below-knee amputation. OUTCOME: The first patient survived, while the second died. CONCLUSION: Traditional medical therapy can cause bacterial inoculation, leading to necrotising fasciitis, and also leads to delay in appropriate treatment. Radical surgery is needed to optimise patient survival. PMID- 16710500 TI - Ethambutol-associated optic neuropathy. AB - INTRODUCTION: Ethambutol is used in the treatment of tuberculosis, which is still prevalent in Southeast Asia, and can be associated with permanent visual loss. We report 3 cases which presented with bitemporal hemianopia. CLINICAL PICTURE: Three patients with ethambutol-associated toxic optic neuropathy are described. All 3 patients had loss of central visual acuity, colour vision (Ishihara) and visual field. The visual field loss had a bitemporal flavour, suggesting involvement of the optic chiasm. TREATMENT: Despite stopping ethambutol on diagnosis, visual function continued to deteriorate for a few months. Subsequent improvement was mild in 2 cases. In the third case, visual acuity and colour vision normalised but the optic discs were pale. OUTCOME: All 3 patients had some permanent loss of visual function. CONCLUSIONS: Ethambutol usage is associated with permanent visual loss and should be avoided if possible or used with caution and proper ophthalmological follow-up. The author postulates that in cases of ethambutol associated chiasmopathy, ethambutol may initially affect the optic nerves and subsequently progress to involve the optic chiasm. PMID- 16710501 TI - Delayed hypersensitivity reaction after intravenous glucagon administered for a barium enema: a case report. AB - INTRODUCTION: Few reports have documented allergic hypersensitivity reactions after barium gastrointestinal studies. Of these, the barium suspension, its additives or intravenous glucagon given for bowel relaxation has been implicated as possible allergens. We report a patient with delayed hypersensitivity reaction after barium enema and discuss the reasons supporting glucagon as the possible allergen. CLINICAL PICTURE: A 74-year-old Chinese woman presented with pruritic rashes, 1 day after a barium enema. Intravenous glucagon (GlucaGen, Novo Nordisk, Denmark) was administered during the barium enema. Physical examination revealed palpable purpuric rashes on the legs with erythematous papules and plaques on the arms and trunk. Skin biopsy demonstrated superficial perivascular infiltrates of lymphocytes and eosinophils, consistent with a drug eruption. TREATMENT AND OUTCOME: The rashes resolved with antihistamines and topical corticosteroids. CONCLUSION: This report highlights the potential of glucagon to cause hypersensitivity reactions. Awareness of this entity is important for the prevention and recognition of complications during barium gastrointestinal studies. PMID- 16710503 TI - [Blockade of iodine in thyroid cancer in known cases. Principles, effectiveness and actual cases]. PMID- 16710502 TI - Methodologies for interventional myopia studies. AB - Myopia studies are notoriously difficult to carry out. Past studies on intervention in myopia progression have given conflicting results. Beside inaccurate and inadequate measurements, the most important cause for this is the very variable nature of myopia, which makes it difficult to achieve baseline comparability between the control and the study group. Although there were inclusion criteria in these studies, for age, sex, race, degree of myopia and stigmatism, the most important variate-- the rate of myopia progression-- was not included. Randomisation can achieve baseline comparability of the myopia progression rate, provided the sample sizes are large enough. Unfortunately, past studies have been limited to 100 to 200 children only. Studies on twins are more reliable than random groups because myopia progression rates are more likely to be the same in a pair of twins. Studies on the same subject, comparing the right eye and the left eye would be even better, but this method is practicable for some studies only (e.g., we cannot have a spectacle lens for one eye and a contact lens on the fellow eye). There is another method of doing an interventional study on myopia. Because myopia progression is linear in its early stage until the early teenage years, it is possible to observe what happens to the linear progression upon intervention. In this way, we avoid the problem of trying to compare "apples with apples" but use the "same apple" instead. PMID- 16710504 TI - [Radioiodine therapy for combined disseminated and nodular thyroid autonomy. Results after using a correction term for the disseminated part]. AB - AIM: In combined focal and disseminated thyroid autonomy a variety of concepts in the treatment with radioiodine are used. The difference lies mainly in the calculation of the autonomous volume. This retrospective study shows a new method of calculating the autonomous volume. PATIENTS AND METHODS: In 398 patients with combined thyroid autonomy and good correlation of scintigraphically hot nodules and lesions defined by ultrasound the volume of the nodules is ascertained from scintigraphic and ultrasound parameters and the volume of the disseminated autonomous tissue is assessed with a weighting factor (VF). This factor is the ratio of impulse density in a ROI over the disseminated volume divided by the corresponding impulse density over the nodular volume of the thyroid scintigraphy. The sum of nodular volume and weighted perinodular volume gives the total autonomous volume. A standard radioiodine test gives the maximum iodine-131 uptake and effective half-life to calculate the activity to obtain a treatment dose of 400 Gy. RESULTS: The rate of success with and without thyrostatic medication was 97% with an 18.6% rate of hypothyroidism observed from 4 months post therapy onwards. CONCLUSION: The use of the weighting factor VF in the treatment of combined autonomy leads to an excellent rate of success in patients with good correlation of functional imaging and ultrasound findings. PMID- 16710505 TI - (18)F-FDG PET and conventional imaging for assessment of Hodgkin's disease and non Hodgkin's lymphoma. An analysis of 193 patient studies. AB - The AIM of this study was to assess the diagnostic value of FDG-PET and conventional imaging (CI) in a large series of patient with Hodgkin's disease (HD) or non-Hodgkin's lymphoma (NHL) at three time points during their course of disease. PATIENTS, METHODS: 169 consecutive lymphoma patients (69 HD; 100 NHL) were included. 193 FDG-PET studies were performed for staging at baseline in 42 cases, for post-therapeutic monitoring in 103, and for diagnosis of recurrence in 48 cases. Performance indices of sensitivity, specificity, positive (PPV) and negative predictive value (NPV), and accuracy of metabolic FDG-PET and morphological CI were calculated. Differences in staging and diagnosis of residual or recurrent lymphoma were compared. RESULTS: FDG-PET changed staging in 36% of cases for staging at baseline, in 52% of cases for monitoring response to treatment, and in 29% for diagnosis of recurrence. FDG-PET staging results were confirmed in 80% for staging at baseline, in 74% for monitoring response to treatment, and in 50% for diagnosis of recurrence. FDGPET and CI differed significantly at monitoring response to treatment for sensitivity (0.91 versus 0.69; p < 0.02), specificity (0.90 versus 0.38; p < 0.00001), PPV (0.77 versus 0.42; p < 0.001), and accuracy (0.83 versus 0.55; p < 0.02). CONCLUSION: FDG-PET should be considered as the diagnostic modality of choice for post-therapeutic assessment of lymphoma patients and may be a reliable alternative to CI for staging at baseline and diagnosis of recurrence. PMID- 16710506 TI - Myocardial perfusion SPECT with dipyridamole stress test in cardiac syndrome X. AB - Cardiac syndrome X defines patients with typical anginal chest pain, a positive exercise ECG stress test and angiographically normal coronary arteries. AIM of this study was to evaluate the role of myocardial perfusion SPECT with dipyridamole stress in the diagnosis of cardiac syndrome X. PATIENTS, METHODS: 68 patients with syndrome X aged 32 to 60 years were subjected to myocardial imaging using (99m)Tc-MIBI according to the two-days protocol: at rest and after dipyridamole infusion. Semiquantitative evaluation of the images was based on the assessment of (99m)Tc-MIBI uptake in 17 myocardial segments using a 5-points scale (0 point -- normal uptake, 4 points -- no uptake). Scores obtained in each segment were summed up, constituting the summed rest score (SRS) and summed stress score (SSS). RESULTS: Mean SRS was 7.9 +/- 4.8 and mean SSS was 7.2 +/- 4.4 (non-significant difference). Individual comparison of SRS and SSS values revealed three patterns of scintigraphic images: 1) in 25 patients (36.8%), a paradoxical improvement of perfusion at stress images was found, 2) in 23 patients (33.8%), the myocardial perfusion deteriorated after dipyridamole, 3) in 20 patients (29.4%), no significant change of the myocardial perfusion between rest and stress images occurred. CONCLUSIONS: In cardiac syndrome X, myocardial SPECT with dipyridamole stress shows different patterns of myocardial perfusion that reflects heterogeneity of this pathology. PMID- 16710507 TI - [Localisation of parathyroid glands using planar (99m)Tc-sestamibi scintigraphy. Comparison between subtraction- and dual-phase technique]. AB - AIM: In the context of presurgical localisation of parathyroid adenomas in primary hyper-parathyreoidism (pHPT) using (99m)Tc-sestamibi scintigraphy, subtraction- and dualphase technique are compared with each other and with the surgical findings. PATIENTS, METHODS: Prospectively, 126 patients with pHPT were investigated presurgically. For visualisation of parathyroid adenomas, an image of the thyroid ((99m)Tc-pertechnetat) was subtracted from a perfusion image ((99m)Tc-sestamibi) and 2 h p. i. another image was acquired for identification of retention of activity. Considering both techniques the clinical findings were reported promptly. Retrospectively, the evaluations were presented separately to four experienced raters. RESULTS: In clinical routine for 109 patients correct findings were reported presurgically (87%). From 129 resected parathyroid adenomas 118 were localised correctly (sensitivity 91%, positive predictive value 94%). Concerning the retrospective analysis, in 75% of the cases both techniques provided the correct site, in 14% only the dual-phase technique and in 7% only the subtraction-technique was correct. With the help of the dual-phase technique significantly more investigations were correctly rated than with the help of the subtraction technique (88.7 +/- 3.2% vs. 81.6 +/- 1.2%, p < 0.01, two-sided t test). CONCLUSION: The presurgical scintigraphic localisation of hyperactive parathyroid glands in pHPT assists minimal invasive surgery serving a high rate of correct findings. According to our data the dual-phase technique seems to be more sensitive than the subtraction technique. In some cases, however, the correct site may only be found using the subtraction technique. For an optimal surgical strategy we suggest the combination of both techniques. PMID- 16710508 TI - Decrease of (99m)Tc-uptake in autonomous thyroid tissue in Germany since the 1970s. Clinical implications for radioiodine therapy. AB - AIM: The clinical relevance of thyroidal autonomy, i.e. the risk of a patient to become hyperthyroid after exposure to iodine, can be estimated by measurement of the thyroidal (99m)Tc uptake under suppression of TSH (TcTUs). The upper tolerable limit has been set to 2% some 25 years ago. Considering the increase in nutritional iodine uptake over the last 15 years, we wanted to find out if the TcTUs per ml of autonomous volume may have changed. PATIENTS, METHODS: We performed a pilot study in 1166 randomly chosen patients from 1980-2003 with different kinds of benign thyroid disorders to determine changes in TcTU or TcTUs over time. A second analysis was performed in 1063 patients from 1987-2004 with unifocal autonomy (UFA). In these patients, the volume of the autonomous tissue can be determined precisely thus allowing for exact determination of TcTUs per ml of autonomous volume. RESULTS: The pilot study demonstrated that the TcTUs or the TcTU has been falling over the last 25 years in all benign thyroid disorders (p < 0.01). The total thyroid volume has also been decreasing in all disorders. In the second analysis of UFA only, 500 from the 1063 patients fulfilled the inclusion criteria. In these patients, the TcTUs per ml of autonomous volume has fallen from an average of 0.48% to an average of 0.28%. These results are statistically significant as determined by ANOVA testing (p = 0.032). CONCLUSION: As the TcTUs in relation to autonomous volume has dropped by approximately 40% over the last 25 years, the upper limit for a normal TcTUs should be reduced to 1-1.4%, dependent on regional factors. PMID- 16710509 TI - PET/CT Biograph Sensation 16. Performance improvement using faster electronics. AB - AIM: The new PET/CT Biograph Sensation 16 (BS16) tomographs have faster detector electronics which allow a reduced timing coincidence window and an increased lower energy threshold (from 350 to 400 keV). This paper evaluates the performance of the BS16 PET scanner before and after the Pico-3D electronics upgrade. METHODS: Four NEMA NU 2-2001 protocols, (i) spatial resolution, (ii) scatter fraction, count losses and random measurement, (iii) sensitivity, and (iv) image quality, have been performed. RESULTS: A considerable change in both PET count-rate performance and image quality is observed after electronics upgrade. The new scatter fraction obtained using Pico-3D electronics showed a 14% decrease compared to that obtained with the previous electronics. At the typical patient background activity (5.3 kBq/ml), the new scatter fraction was approximately 0.42. The noise equivalent count-rate (R(NEC)) performance was also improved. The value at which the R(NEC) curve peaked, increased from 3.7 x 10(4) s(-1) at 14 kBq/ml to 6.4 x 10(4) s(-1) at 21 kBq/ml (2R-NEC rate). Likewise, the peak true count-rate value increased from 1.9 x 10(5) s(-1) at 22 kBq/ml to 3.4 x 10(5) s(-1) at 33 kBq/ml. An average increase of 45% in contrast was observed for hot spheres when using AW-OSEM (4ix8s) as the reconstruction algorithm. For cold spheres, the average increase was 12%. CONCLUSION: The performance of the PET scanners in the BS16 tomographs is improved by the optimization of the signal processing. The narrower energy and timing coincidence windows lead to a considerable increase of signal- to-noise ratio. The existing combination of fast detectors and adapted electronics in the BS16 tomographs allow imaging protocols with reduced acquisition time, providing higher patient throughput. PMID- 16710511 TI - [Integration of PACS and HIS into the workflow of a nuclear medicine department. Experience in Regensburg]. AB - AIM: The development of new diagnostic techniques and the implementation of a modern quality control management system requires the continuous adaptation of existing data processing tools to the nuclear medicine diagnostic workflow. Furthermore, PACS connected to HIS facilitates and enhances the transfer of data and pictures, and satisfies the legal requirements for data retention as regulated by law. Therefore, the aim of this work is to present the architecture, structure and results of such a system newly installed in a department of nuclear medicine. METHODS: Initially, the nuclear medicine workflow was carefully analyzed and each step was correlated to the corresponding module. The standard SAP R/3 and IS-H/IS-H(*)med based software used for patient administration at the University of Regensburg Hospital was adapted to the needs of the Nuclear Medicine Department. The networking of the imaging systems was done by integration of a PACS. Finally, the PACS was connected to the HIS to allow the attachment of images to the medical report. RESULTS, CONCLUSION: By connecting the HIS to the nuclear medicine PACS, the workflow was significantly improved. The data management sequence starting at the reception desk, continuing through the nuclear medical examination, to the physician's final written and image report is clearly structured. Although high demands exist on technical support and administration the integration of PACS and HIS into the nuclear medicine workflow leads to enhanced efficiency and reduction in hospital costs. Patient and data management are considerably improved in this way. PMID- 16710510 TI - Dosimetry with (188)Re-labelled monoclonal anti-CD66 antibodies. A simplified approach based on a single measurement 3 h p.i. AB - AIM: For the therapeutic application of radiopharmaceuticals the activity is determined on an individual basis. Here we investigated the accuracy for a simplified assessment of the residence times for a (188)Re-labelled anti-CD66 monoclonal antibody. PATIENTS, METHODS: For 49 patients with high risk leukaemia (24 men, 25 women, age: 44 +/- 12 years) the residence times were determined for the injected (188)Re-labelled anti-CD66 antibodies (1.3 +/- 0.4 GBq, 5-7 GBq/mg protein, >95% (188)Re bound to the antibody) based on 5 measurements (1.5, 3, 20, 26, and 44 h p.i.) using planar conjugate view gamma camera images (complete method). In a simplified method the residence times were calculated based on a single measurement 3 h p.i. RESULTS: The residence times for kidneys, liver, red bone marrow, spleen and remainder of body for the complete method were 0.4 +/- 0.2 h, 1.9 +/- 0.8 h, 7.8 +/- 2.1 h, 0.6 +/- 0.3 h and 8.6 +/- 2.1 h, respectively. For all organs a linear correlation exists between the residence times of the complete method and the simplified method with the slopes (correlation coefficients R > 0.89) of 0.89, 0.99, 1.23, 1.13 and 1.09 for kidneys, liver, red bone marrow, spleen and remainder of body, respectively. CONCLUSION: The proposed approach allows reliable prediction of biokinetics of (188)Re-labelled anti-CD66 monoclonal antibody biodistribution with a single study. Efficient pretherapeutic estimation of organ absorbed dose may be possible, provided that a more stable anti-CD66 antibody preparation is available. PMID- 16710512 TI - Cytogenetics of two sympatric Corydoras species (pisces, siluriformes, challichtyidae) of Southern Brazil. AB - Karyotypic data are presented for two sympatric Corydoras species of the Lagoa Dourada, namely, C. ehrhadti and C. paleatus, which are found in the upper Tibagi river basin (Ponta Grossa, State of Parana, Brazil). The same diploid number and karyotypic formula were observed in both species/populations. A great similarity in the constitutive heterochromatin distribution and in the activity of nucleolar organizer regions was also found. The use of in situ hybridization with a fluorescent 18S rDNA probe allowed for the identification of the species/populations through the location of ribosomal sites. PMID- 16710513 TI - Behavior of Angiostrongylus costaricensis in planorbids. AB - Some terrestrial mollusks are natural hosts of Angiostrongylus costaricensis. In the laboratory, this nematode can be maintained in certain planorbids, which are aquatic mollusks and intermediate hosts of Schistosoma mansoni. Mollusks can be infected with Angiostrongylus costaricensis by ingestion of or active penetration by the first-stage larvae. In this work we assessed the ability of Biomphalaria glabrata to attract first-stage larvae of A. costaricensis. Movement of the nematode larvae towards the mollusks was observed after 15 min, 30 min and 1 h. B. glabrata did not attract the first-stage larvae of A. costaricensis in any of the three intervals. The susceptibility of two populations of Biomphalaria tenagophila to infection by A. costaricensis was also determined. One population was genetically selected for the susceptibility to S. mansoni while the other was not. Third-stage larvae were recovered from the snails 30 days after exposure of the two populations to 120 first-stage larvae. All the mollusks were infected. However, a significantly higher number of third-stage larvae were recovered in mollusks not genetically selected. PMID- 16710514 TI - A new sympatric region for distinct karyotypic forms of Hoplias malabaricus (Pisces, Erythrinidae). AB - Specimens of Hoplias malabaricus from Lagoa Carioca, an isolated lake of the Rio Doce State Park (state of Minas Gerais, Brazil), were cytogenetically studied. The diploid number was found to be constant, i.e., 2n = 42 chromosomes, although two karyotypic forms were found: karyotype A, characterized by 22M + 20SM chromosomes, observed only in a male specimen, and karyotype B, characterized by 24M + 16SM + 2ST and 24M + 17SM + 1ST chromosomes in female and male specimens, respectively. This sex difference found in karyotype B is related to an XX/XY sex chromosome system. Another female specimen of H. malabaricus, also carrying karyotype A, had previously been found in the same lake. The available data indicate that two sympatric cytotypes of H. malabaricus exist in the Lagoa Carioca, with cytotype A occurring at a lower frequency and differing from cytotype B by undifferentiated sex chromosomes. PMID- 16710515 TI - Occurrence of toxin-producing cyanobacteria blooms in a Brazilian semiarid reservoir. AB - We report the occurrence of cyanobacterial blooms and the presence of cyanotoxins in water samples from the Armando Ribeiro Goncalves reservoir (06 degrees 08 S and 37 degrees 07 W), located in the state of Rio Grande do Norte, in the semiarid region of northeastern Brazil. The cyanobacterial species were identified and quantified during the rainy and dry seasons in the year 2000. Cyanotoxins such as microcystins, saxitoxins and cylindrospermopsins were analyzed and quantified using HPLC and ELISA methods. The mixed toxic blooms of Cylindrospermopsis raciborskii, Microcystis spp (M. panniformis, M. protocystis, M. novacekii) and Aphanizomenon spp (Aphanizomenon gracile, A. cf. manguinii, A. cf. issastschenkoi) were persistent and represented 90-100% of the total phytoplankton species. Toxic cyanobacterial blooms from the Armando Ribeiro Goncalves reservoir were analyzed and found to have three phases in relation to the annual cycle. During the rainy season, an intense toxic bloom of Cylindrospermopsis raciborskii was recorded along with saxitoxins (3.14 microg.L( 1)). During the transition period, between the rainy and dry seasons, different species of Microscytis occurred and microcystin as high as 8.8 microg.L(-1) was recorded. In the dry season, co-dominance of Cylindrospermopsis raciborskii, Microcystis spp and Aphanizomenon spp occurred and the concentrations of saxitoxin remained very low. Our results indicate the presence of microcystins (8.8 microg.L(-1)) and saxitoxins (3.14 microg.L(-1)) into the crude water, with increasing concentrations from the second fortnight of April to late May 2000. The occurrence of toxic blooms in this reservoir points to a permanent risk of cyanotoxins in supply waters, indicating the need for the implementation of bloom control measures to improve the water quality. Exposure of the local population to cyanotoxins through their potential accumulation in fish muscle must also be considered. PMID- 16710516 TI - Seasonal dynamics of microzooplankton in the Sao Sebastiao Channel (SP, Brazil). AB - The qualitative composition, numerical abundance and seasonal distribution of microzooplankton in the Sao Sebastiao Channel were studied in different seasonal periods. Data are presented for 43 Protozoa, 4 rotifer and 30 copepod nauplii specimens. Tintinnids represented the most abundant group within the microzooplankton community, with 31 species recorded, among which Amphorellopsis acuta, Codonellopsis morchella, Eutintinnus tenuis, Favella ehrenbergii, Leprotintinnus nodqvisti, Tintinnopsis buetschlil, T. lobiancoi and T. radix stood out, followed in abundance by copepod nauplii. Our findings indicated that water masses play a fundamental role in the seasonal composition of microzooplankton assemblages in the Sao Sebastiao Channel. PMID- 16710518 TI - Annual and short-term variability in primary productivity by phytoplankton and correlated abiotic factors in the Jurumirim Reservoir (Sao Paulo, Brazil). AB - The annual variability of the photosynthetic production (PP) by phytoplankton in the lacustrine zone of the Jurumirim Reservoir (Sao Paulo, Brazil) was evaluated in a three-year study to identify recurrent patterns and their causes. Variability in PP was measured daily during two periods of the year (the dry and rainy seasons). An analysis of the PP data failed to identify a recurrent pattern, since the PP values showed no correlation with hydrological factors (rainfall, water level and discharge, and washout) nor, apparently, with the waters nutritional conditions. A principal component analysis revealed that the PP and assimilation ratio were higher when the PO4(3-) and N-NH(4+) contents were low and the ZEU/ZMIX ratios were at their highest. Areal primary productivity can be predicted based on the ratio between the maximum volumetric productivity and the coefficient of vertical extinction of light. However, the biomass integrated for ZEU was a poor predictor of areal primary productivity. No correlation was found between water temperature and areal and maximum volumetric productivity. Thus, the three-year PP study indicated that the variability pattern is typically chaotic. As for the short-term measurements, the PP was found to be higher in the dry season than in the rainy, although both seasons showed an areal PP variability of 35 to 40%. This pattern was attributed to the daily variation in the nutritional conditions and the magnitude of light penetrating through the water, combined with the mixing of phytoplanktonic cells. A comment about the relationship between primary production by phytoplankton and fish yield is also briefly discussed here. PMID- 16710517 TI - Social isolation and aggressiveness in the Amazonian juvenile fish Astronotus ocellatus. AB - We tested the effect of social isolation on the aggressiveness of an Amazonian fish: Astronotus ocellatus. Ten juvenile fishes were transferred from a group aquarium (60 x 60 x 40 cm) containing 15 individuals (without distinguishing sex) to an isolation aquarium (50 x 40 x 40 cm). Aggressiveness was tested by means of attacks on and displays toward the mirror image. The behavior was video-recorded for 10 min at a time on 4 occasions: at 30 min, 1 day, 5 days and 15 days after isolation. The aggressive drive was analyzed in three ways: latency to display agonistic behavior, frequency of attacks and specific attacks toward the mirror image. The latency to attack decreased during isolation, but the frequency of mouth fighting (a high aggressive attack) tended to increase, indicating an augmented aggressive drive. Our findings are congruent with the behavior of the juvenile cichlid, Haplochromis burtoni but differ from the behavior observed in another cichlid, Pterophylum scalare. Increased aggressiveness in A. ocellatus may be mediated by means of the primer effect, the effect of prior residence or processes involving recognition of a conspecific. PMID- 16710519 TI - Stock assessment and fishery management of the pink shrimp Farfantepenaeus brasiliensis Latreille, 1970 and F. paulensis Perez-Farfante, 1967 in Southeastern Brazil (23 degrees to 28 degrees S). AB - Quantitative analyses of the pink shrimp (Farfantepenaeus brasiliensis and Farfantepenaeus paulensis) fisheries were carried out using data collected from July 1999 to July 2001 from the trawling operations of the fishing fleet based in Santos/Guaruja, SP. According to classical models, the fishery is at its maximum sustainable yield. Therefore, reduction of the fishing effort and adequate season and area closures seem to be the best management actions for the pink shrimp fishery. PMID- 16710520 TI - Reproductive relationships and degree of synapsis in the polytene chromosomes of the Drosophila buzzatii species cluster. AB - The process of speciation occurs through the evolution of any of several forms of reproductive isolation between taxa, including inviability of hybrids. In this work, strains derived from allopatric populations of Drosophila buzzatii cluster species were experimentally crossed in order to evaluate their reproductive and cytogenetic relationships, and to contribute toward understanding the reproductive isolation in this group of sibling species. Although intrastrain crosses were highly fertile, we consider it relevant to discuss the differences in intra- and interspecific fertility and fecundity here. Among 30 interspecific crosses, about 63% were partially or completely sterile. Fifty three percent of interspecific F1 crosses (female and male F1 crossed) were also partially or completely sterile, in contrast to only one out of 24 intraspecific F1 crosses that was partially sterile. An analysis of hybrid polytene chromosomes revealed complete synapsis, except in the microchromosomes (VI) and in the proximal region of the X chromosome. The intraspecific divergence observed in this study and the variable degree of chromosome pairing shown here reveal part of the complexity of the speciation process pertinent to Drosophila buzzatii cluster, which is consistent with different traits studied in this cluster. PMID- 16710521 TI - First record of the bat Mimon crenulatum (E. Geoffroy, 1801) (Mammalia: Chiroptera) in the state of Rio de Janeiro, Southeastern Brazil. AB - The present study reports an extension of the geographic range of the phyllostomid bat Mimon crenulatum. This is the first record of this species in the state of Rio de Janeiro, Southeastern Brazil. Bats were captured in two conservation units of the Atlantic Forest. Data on the ecology and morphometry of the individuals are presented and compared with data recorded for other localities. The occurrence of this bat species in the region, though new, is consistent with information on its natural history found in the literature. PMID- 16710522 TI - Colleters in Caryocar brasiliense (Caryocaraceae) ontogenesis, ultrastructure and secretion. AB - Colleters are widely distributed and have been reported in several taxa. However, the only report of colleters in the Caryocaraceae family is in Caryocar brasiliense. This study purports to describe the distribution, structure and ultrastructure of colleters in C. brasiliense. Vegetative shoot apices with stipules in different stages of development were collected, fixed, and processed by light, transmission and scanning electron microscopy. Colleters occur on the stipule adaxial face and are differentiated on younger stipules, remaining functional until senescence. Stipules are deciduous, falling before the beginning of leaf expansion. Colleter secretion is abundant, filling all the space delimited by the stipule. Colleter origin is mixed, involving the protoderm and ground meristem. In this species, colleters are finger-shaped, consisting of a column of non-secretory central cells and a secretory epithelium whose cells are arranged in palisades. Secretory epithelium cells show dense cytoplasm, rough endoplasmic reticulum (RER) segments, free ribosomes, mitochondria, and abundant well developed dictyosomes. The colleters central cell shows a flocculated cytoplasm and developed vacuole. These cells have a small lobated nucleus with an inconspicuous nucleolus. Mitochondria and RER stand out in the organelles. There is evidence of granulocrine secretion and colleter involvement with protection of the apical meristem and leaves in the initial stages of development. PMID- 16710523 TI - Ultrastructural aspects of the intercellular bridges between female bee germ cells. AB - The germline cells in the ovary of the female bee are interconnected by intercellular bridges kept open by cytoskeletal reinforcements in the plasmic membrane. These bridges among the germline cells display a dynamic behavior and probably act in the determination of the oocyte among the cells of the clone formed by the premeiotic mitoses, subsequently forming a pathway that enables the products synthesized by the nurse cells to reach the oocyte during its maturation. The cytoskeletal elements in the intercellular bridges of bee gonads are basically microfilaments and microtubules, but another type of filament (thick, of non-defined nature, associated with elements of the endoplasmic reticulum) is present in the bridges between the premeiotic cystocytes. This filament crosses the bridge, using microfilaments to fasten itself to the plasmic membrane. These filaments appear to control the span of the bridge. Upon completion of the proliferation phase the cystocytes take on a rosette shape, and a fusome formed by the convergence of the bridges appears at their center. The thick filaments are not present in this conformation. The differentiation of the oocyte and the nurse cells leads to a new change, in which the bridges are reoriented to convey the content of the future nurse cells to the oocyte. PMID- 16710524 TI - First gonadal maturation of Pinirampus pirinampu (Siluriformes: Pimelodidae) in the Pantanal, Mato Grosso do Sul State, Brazil. AB - Information regarding the reproductive cycle of commercially exploited fish species is extremely important for their appropriate management. The reproductive size of Pinirampus pirinampu (Spix, 1829) was determined for a population studied in the Pantanal wetlands of western Brazil. Samples were taken in September, October, and December 1997, and in February and March 1998. Gonadal stages were macroscopically determined and a monthly gonadossomatic index (GSI) was used to determine the breeding period. The length of the first gonadal maturation was determined by classifying male and female adults and immature fish into length classes. The data on the adult individuals were plotted on graphs and the median values estimated to find the length at which 50% of the individuals reached maturity (L50). L100 was also determined. Males and females in the gonadal maturation process were recorded from October on, and the highest frequency was found to occur in February. Individuals with empty gonads occurred from February onwards. The GSI showed that gonadal development began in September and reached its peak in February. The L50 was 574 mm for females and 536 mm for males, while the L100 was 590 mm for females and 580 mm for males. PMID- 16710525 TI - Physiological rates in different classes of sizes of Perna perna (Linnaeus, 1758) submitted to experimental laboratory conditions. AB - Physiological studies of the mussel Perna perna in Brazil are almost 30 years behind those of other, more exhaustively investigated species, such as Mytilus edulis. Little is known about the variations in physiological rates due to size and the consequences of maintaining P. perna in laboratory conditions. This work investigated the variations in respiration, clearance, excretion and absorption efficiency rates of P. perna, classified by size and acclimatized in a laboratory, monitoring the mussels respiration rates and biometry over a period of 30 days, in laboratory conditions. The respiration, clearance and excretion rates presented an allometric relation with the dry weight of the organisms, with b values of 0.66, 0.48 and 0.91 respectively. On the other hand, these same rates, when considered by weight (specific rates) showed a relationship that was inverse to the size of the organisms. Only the absorption efficiency was independent of the weight of the mussel. In terms of acclimatization, it was observed that it takes 10 days for the respiration rate of the mussel P. perna to stabilize in laboratory conditions, after which it follows a routine metabolism. PMID- 16710526 TI - Zooplankton biomass of reservoirs in different trophic conditions in the State of Sao Paulo, Brazil. AB - This paper reports on a study involving an estimate of the biomass of rotifers, cladocerans, and cyclopoid and calanoid copepods found in reservoirs in different trophic conditions, comparing and relating numerical density data and pointing out differences between the dry and rainy seasons. In terms of numerical densities, both reservoirs were dominated by rotifers, although cladocerans represented a higher biomass in the oligotrophic environment in both seasons. In the eutrophic environment, higher biomass values were found for cyclopoids during the dry season and for cladocerans during the rainy one. Different biomass patterns were observed relating to both the trophic conditions and the rainy and dry seasons. PMID- 16710527 TI - Microhabitat use by two rocky shore gastropods in an intertidal sandy substrate with rocky fragments. AB - Sandy beaches in some areas of the Sao Sebastiao Channel in southeastern Brazil have unremittingly undergone a variety of impacts, including the deposition of rock fragments in the intertidal region. Consequently, these environments support a rich fauna comprising both sandy beach and rocky shore organisms. Two rocky shore gastropods, Tegula viridula and Morula nodulosa, are particularly abundant in such environments. An evaluation of the use of microhabitats by these two species revealed that they occupy the available microhabitats in different proportions and the presence of one species is associated with the absence of the other. Morula nodulosa is randomly dispersed, occupying mostly areas with rock fragments covered with sediment and branching brown algae. Tegula viridula shows a clumped dispersion associated with the patchiness of the microhabitats used: the presence of encrusting green algae and absence of sediment and branching brown algae covering the rocks. These findings suggest T. viridula has a lower tolerance than M. nodulosa to sand inundation of the rocky fragments, a stochastic event common to the environment in question. PMID- 16710528 TI - Galls and gall makers in plants from the Pe-de-Gigante Cerrado Reserve, Santa Rita do Passa Quatro, SP, Brazil. AB - Thirty-six morphologically different types of galls were obtained in leaves, leaflets, veins, petioles, stems, tendrils and flower buds from twenty-five species of plants in the Pe-de-Gigante Reserve, municipality of Santa Rita do Passa Quatro, state of Sao Paulo, Brazil. The host plant species belong to the closely related families Anacardiaceae, Annonaceae, Asteraceae, Bignoniaceae, Caryocaraceae, Erythroxylaceae, Fabaceae, Malpighiaceae, Melastomataceae, Myrtaceae, Ochnaceae, Polygalaceae, Sapindaceae, Sapotaceae, and Smilacaceae. The most common gall makers included Cecidomyiidae (Diptera), Pteromalidae (Hymenoptera) and Diaspididae (Sternorrhyncha-Hemiptera). This is the first report of galls found in the following plant genera: Gochnatia (Asteraceae), Distictela (Bignoniaceae), Banisteriopsis (Malpighiaceae), Ouratea (Ochnaceae), and Bredemeyera (Polygalaceae). The results of this work contribute to the body of knowledge about the relationship among host plants, gall makers, and the gall morphology of Pe-de-Gigante Cerrado Reserve. PMID- 16710529 TI - Capybaras in an anthropogenic habitat in Southeastern Brazil. AB - Capybaras (Hydrochoerus hydrochaeris) are widely distributed in the Neotropics, living in both natural and anthropogenic habitats. In the present study, we monitored the fluctuating capybara population at the Campus "Luiz de Queiroz", University of Sao Paulo, in Piracicaba, Southeastern Brazil from 1998 to 2000. To assess population fluctuation and growth rates, we used an abundance index based on direct weekly daylight counts. Population monitoring was carried out in an anthropogenic wetland associated with an agroecosystem. The observers bias was established as directly related to their distance from the animals. The capybara population density and biomass in the anthropogenic wetland in question were found to be significantly higher than in pristine habitats. The species seems to present a seasonal pattern of fluctuation, with a peak in late spring (Oct to Dec). Unlike young capybaras, adults and juveniles present similar seasonal fluctuation patterns. In this anthropogenic wetland, the carrying capacity may be as high as 195 individuals/Km2. At a conservative exploitation rate (17%) in anthropogenic wetlands of Southeastern Brazil, capybara productivity may reach approximately 630 Kg/Km2/year. PMID- 16710530 TI - Occurrence of helminths in bristle-spined porcupine (Chaetomys subspinosus) (Olfers, 1818), Salvador, Brazil. PMID- 16710531 TI - Handling technique to increase the hyacinth macaw population (Anodorhynchus hyacinthinus) (Lalham, 1720): report of an experience in Pantanal, Brazil. PMID- 16710532 TI - Public health, innovation and intellectual property rights: unfinished business. PMID- 16710533 TI - The impact of open access upon public health. PMID- 16710534 TI - Intellectual property and public health. PMID- 16710535 TI - Public health ethics and intellectual property policy. PMID- 16710536 TI - Access to AIDS medicines stumbles on trade rules. PMID- 16710537 TI - Rich and poor countries divided on patent treaty. PMID- 16710538 TI - Protecting traditional knowledge: the San and hoodia. PMID- 16710539 TI - Meeting the need for treatment: the initiatives. PMID- 16710540 TI - Do patents work for public health? PMID- 16710542 TI - CIPIH report: main recommendations. PMID- 16710543 TI - A clearing house for diagnostic testing: the solution to ensure access to and use of patented genetic inventions? AB - In genetic diagnostics, the emergence of a so-called "patent thicket" is imminent. Such an overlapping set of patent rights may have restrictive effects on further research and development of diagnostic tests, and the provision of clinical diagnostic services. Currently, two models that may facilitate access to and use of patented genetic inventions are attracting much debate in various national and international fora: patent pools and clearing houses. In this article, we explore the concept of clearing houses. Several types of clearing houses are identified. First, we describe and discuss two types that would provide access to information on the patented inventions: the information clearing house and the technology exchange clearing house. Second, three types of clearing houses are analysed that not only offer access to information but also provide an instrument to facilitate the use of the patented inventions: the open access clearing house, the standardized licences clearing house and the royalty collection clearing house. A royalty collection clearing house for genetic diagnostic testing would be the most comprehensive as it would serve several functions: identifying patents and patent claims essential to diagnostic testing, matching licensees with licensors, developing and supplying standardized licences, collecting royalties, monitoring whether users respect licensing conditions, and providing dispute resolution services such as mediation and arbitration. In this way, it might function as an effective model for users to facilitate access to and use of the patented inventions. However, it remains to be seen whether patent holders with a strong patent portfolio will be convinced by the advantages of the royalty collection clearing house and be willing to participate. PMID- 16710544 TI - Managing the effect of TRIPS on availability of priority vaccines. AB - The stated purpose of intellectual property protection is to stimulate innovation. The Agreement on Trade-Related Aspects of Intellectual Property Rights (TRIPS) requires all Members of the World Trade Organization (WTO) to enact national laws conferring minimum standards of intellectual property protection by certain deadlines. Critics of the Agreement fear that such action is inconsistent with ensuring access to medicines in the developing world. A WHO convened meeting on intellectual property rights and vaccines in developing countries, on which this paper is based, found no evidence that TRIPS has stimulated innovation in developing market vaccine development (where markets are weak) or that protection of intellectual property rights has had a negative effect on access to vaccines. However, access to future vaccines in the developing world could be threatened by compliance with TRIPS. The management of such threats requires adherence of all countries to the Doha Declaration on TRIPS, and the protections guaranteed by the Agreement itself, vigilance on TRIPS plus elements of free trade agreements, developing frameworks for licensing and technology transfer, and promoting innovative vaccine development in developing countries. The role of international organizations in defining best practices, dissemination of information, and monitoring TRIPS impact will be crucial to ensuring optimal access to priority new vaccines for the developing world. PMID- 16710545 TI - Benchmarking progress in tackling the challenges of intellectual property, and access to medicines in developing countries. AB - The impact of intellectual property protection in the pharmaceutical sector on developing countries has been a central issue in the fierce debate during the past 10 years in a number of international fora, particularly the World Trade Organization (WTO) and WHO. The debate centres on whether the intellectual property system is: (1) providing sufficient incentives for research and development into medicines for diseases that disproportionately affect developing countries; and (2) restricting access to existing medicines for these countries. The Doha Declaration was adopted at WTO in 2001 and the Commission on Intellectual Property, Innovation and Public Health was established at WHO in 2004, but their respective contributions to tackling intellectual property related challenges are disputed. Objective parameters are needed to measure whether a particular series of actions, events, decisions or processes contribute to progress in this area. This article proposes six possible benchmarks for intellectual property-related challenges with regard to the development of medicines and ensuring access to medicines in developing countries. PMID- 16710546 TI - Essential medicines and human rights: what can they learn from each other? AB - Most countries have acceded to at least one global or regional covenant or treaty confirming the right to health. After years of international discussions on human rights, many governments are now moving towards practical implementation of their commitments. A practical example may be of help to those governments who aim to translate their international treaty obligations into practice. WHO's Essential Medicines Programme is an example of how this transition from legal principles to practical implementation may be achieved. This programme has been consistent with human rights principles since its inception in the early 1980s, through its focus on equitable access to essential medicines. This paper provides a brief overview of what the international human rights instruments mention about access to essential medicines, and proposes five assessment questions and practical recommendations for governments. These recommendations cover the selection of essential medicines, participation in programme development, mechanisms for transparency and accountability, equitable access by vulnerable groups, and redress mechanisms. PMID- 16710547 TI - Choosing the right incentive strategy for research and development in neglected diseases. AB - For the first time in history, worldwide neglected disease budgets may be large enough to deliver a new drug every few years. That said, sponsors will only succeed if they extract maximum value from every dollar spent. This paper reviews possible cost-containment strategies and provides an evidence-based framework for choosing between them. Current proposals can be categorized as "end-to-end" proposals which require the sponsor to set a single reward for companies that complete the entire drug discovery process or "pay-as-you-go" schemes in which sponsors offer repeated rewards as drug candidates progress through the pipeline. A generic weakness of end-to-end proposals is that rewards are likely to be 20 30% higher than they would be in an equivalent pay-as-you-go programme. However, the benefits of pay-as-you-go programmes may be lost if commercial pharmaceutical companies are substantially better at choosing successful programmes than are their non-profit counterparts. The efficiency of pay-as-you-go methods depends on sponsors' willingness to withdraw funding from failed drug discovery programmes. PMID- 16710549 TI - DNA patenting: implications for public health research. AB - I weigh the arguments for and against the patenting of functional DNA sequences including genes, and find the objections to be compelling. Is an outright ban on DNA patenting the right policy response? Not necessarily. Governments may wish to consider options ranging from patent law reforms to the creation of new rights. There are alternative ways to protect DNA sequences that industry may choose if DNA patenting is restricted or banned. Some of these alternatives may be more harmful than patents. Such unintended consequences of patent bans mean that we should think hard before concluding that prohibition is the only response to legitimate concerns about the appropriateness of patents in the field of human genomics. PMID- 16710548 TI - Data sharing and intellectual property in a genomic epidemiology network: policies for large-scale research collaboration. AB - Genomic epidemiology is a field of research that seeks to improve the prevention and management of common diseases through an understanding of their molecular origins. It involves studying thousands of individuals, often from different populations, with exacting techniques. The scale and complexity of such research has required the formation of research consortia. Members of these consortia need to agree on policies for managing shared resources and handling genetic data. Here we consider data-sharing and intellectual property policies for an international research consortium working on the genomic epidemiology of malaria. We outline specific guidelines governing how samples and data are transferred among its members; how results are released into the public domain; when to seek protection for intellectual property; and how intellectual property should be managed. We outline some pragmatic solutions founded on the basic principles of promoting innovation and access. PMID- 16710550 TI - The pricing and procurement of antiretroviral drugs: an observational study of data from the Global Fund. AB - The Purchase price report released in August 2004 by the Global Fund to Fight AIDS, Tuberculosis, and Malaria (Global Fund) was the first publication of a significant amount of real transaction purchase data for antiretrovirals (ARVs). We did an observational study of the ARV transaction data in the Purchase price report to examine the procurement behaviour of principal recipients of Global Fund grants in developing countries. We found that, with a few exceptions for specific products (e.g. lamivudine) and regions (e.g. eastern Europe), prices in low-income countries were broadly consistent or lower than the lowest differential prices quoted by the research and development sector of the pharmaceutical industry. In lower middle-income countries, prices were more varied and in several instances (lopinavir/ritonavir, didanosine, and zidovudine/lamivudine) were very high compared with the per capita income of the country. In all low- and lower middle-income countries, ARV prices were still significantly high given limited local purchasing power and economic strength, thus reaffirming the need for donor support to achieve rapid scale-up of antiretroviral therapy. However, the price of ARVs will have to decrease to render scale-up financially sustainable for donors and eventually for governments themselves. An important first step in reducing prices will be to make available in the public domain as much ARV transaction data as possible to provide a factual basis for discussions on pricing. The price of ARVs has considerable implications for the sustainability of human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) treatment in the developing world. PMID- 16710552 TI - A human rights approach to the WHO Model List of Essential Medicines. AB - Since the first WHO Model List of Essential Medicines was adopted in 1977, it has become a popular tool among health professionals and Member States. WHO's joint effort with the United Nations Committee on Economic, Social and Cultural Rights has resulted in the inclusion of access to essential medicines in the core content of the right to health. The Committee states that the right to health contains a series of elements, such as availability, accessibility, acceptability and quality of health goods, services and programmes, which are in line with the WHO statement that essential medicines are intended to be available within the context of health systems in adequate amounts at all times, in the appropriate dosage forms, with assured quality and information, and at a price that the individual and the community can afford. The author considers another perspective by looking at the obligations to respect, protect and fulfil the right to health undertaken by the states adhering to the International Covenant of Economic, Social and Cultural Rights (ICESCR) and explores the relationship between access to medicines, the protection of intellectual property, and human rights. PMID- 16710551 TI - Implications of bilateral free trade agreements on access to medicines. AB - The TRIPS Agreement of the World Trade Organization (WTO) mandated the introduction of protection of intellectual property rights, notably patents, for pharmaceutical products. While the implications for the access to medicines contained in the terms of this Agreement raised significant concerns, a recent new wave of free trade agreements, negotiated outside the WTO, requires even higher levels of intellectual property protection for medicines than those mandated by that Agreement. The measures involved include the extension of the patent term beyond 20 years; prohibition of use of test data on drug efficacy and safety for certain periods for the approval of generic products; the linkage between drug registration and patent protection; in some cases, limitations to the grounds for granting compulsory licences. This article reviews some of these measures that further limit the competition of generic products and discusses their possible implication for access to medicines. PMID- 16710554 TI - Staking claims in the biotechnology Klondike. PMID- 16710555 TI - Expanding global research and development for neglected diseases. PMID- 16710556 TI - Patent policy and public health in developing countries: lessons from Japan. PMID- 16710557 TI - Report of the Commission on Intellectual Property Rights, Innovation and Public Health: an industry perspective. PMID- 16710558 TI - Report of the Commission on Intellectual Property Rights, Innovation and Public Health: a call to governments. PMID- 16710559 TI - The structure of weakly stable minimal hypersurfaces. AB - In this short communication, we announce results from our research on the structure of complete noncompact oriented weakly stable minimal hypersurfaces in a manifold of nonnegative sectional curvature. In particular, a complete oriented weakly stable minimal hypersurface in Rm, m > or = 4, must have only one end; any complete noncompact oriented weakly stable minimal hypersurface has only one end if the complete oriented ambient manifold Nm, m > or = 7, has nonnegative sectional curvature and Ricci curvature bounded below by a positive constant; a complete oriented weakly stable minimal hypersurface in Rm, m > or = 4, with finite total scalar curvature is a hyperplane. PMID- 16710560 TI - Aedeagus morphology as a discriminant marker in two closely related cactophilic species of Drosophila (Diptera; Drosophilidae) in South America. AB - Drosophila serido and D. antonietae are sibling species belonging to the Drosophila buzzatii cluster. Morphologically, they can only be discriminated by quantitative traits. In this paper we analyze the length and equalized average curvature of four regions of the aedeagus of D. antonietae and D. serido. Specimens of D. serido and D. antonietae were classified correctly 96.74% of the time. Based only on the variable that most contributed to the discrimination of the groups (equalized average curvature of the arch IV of the aedeagus), we observed significant intraspecific morphological divergence in D. serido in relation to the D. antonietae, in agreement with other markers. The high morphological divergence in equalized average curvature of the arch IV of the aedeagus shows that this region evolved faster than others, since the divergence of the two species. The importance of the present study to the understanding of the genetic basis that controls the formation of the aedeagus, in the species of the Drosophila buzzatii cluster, is discussed. PMID- 16710561 TI - Rhizophores in Rhizophora mangle L: an alternative interpretation of so-called ''aerial roots''. AB - Rhizophora mangle L., one of the most common mangrove species, has an aerial structure system that gives it stability in permanently swampy soils. In fact, these structures, known as "aerial roots" or "stilt roots", have proven to be peculiar branches with positive geotropism, which form a large number of roots when in contact with swampy soils. These organs have a sympodial branching system, wide pith, slightly thickened cortex, collateral vascular bundles, polyarch stele and endarch protoxylem, as in the stem, and a periderm produced by a phellogen at the apex similar to a root cap. They also have the same type of trichosclereid that occurs in the stem, with negative geotropism, unlike true Rhizophora roots, which do not form trichosclereids at all. On the other hand, these branches do not form leaves and in this respect they are similar to roots. These peculiar branches are rhizophores or special root-bearing branches, analogous to those found in Lepidodendrales and other Carboniferous tree ferns that grew in swampy soils. PMID- 16710562 TI - Paleoenvironment interpretation of a 1760 years B.P. old sediment in a mangrove area of the Bay of Guanabara, using pollen analysis. AB - A sediment sample was obtained at 122 cm from the top of a drilling core in the Guapimirim mangrove, Bay of Guanabara, and analyzed using pollen analysis. This muddy core reached a sandy ground at 133 cm. 14C datation got the age of 1760 +/- 50 years B.P. The most frequent pollen grains were mangrove species of Rhizophora mangle, Laguncularia racemosa and Avicennia schaueriana. "Restinga" and tropical rain forest vegetation was recognized behind the mangrove. After the last sea transgression at 2500 years B.P., the water level lowered to its actual size, allowing the installation of this mangrove. PMID- 16710564 TI - The structural molecular biology network of the State of Sao Paulo, Brazil. AB - This article describes the achievements of the Structural Molecular Biology Network (SMolBNet), a collaborative program of structural molecular biology, centered in the State of Sao Paulo, Brazil, and supported by Sao Paulo State Funding Agency (FAPESP). It gathers twenty scientific groups and is coordinated by the scientific staff of the Center of Structural Molecular Biology, at the National Laboratory of Synchrotron Light (LNLS), in Campinas. The SMolBNet program has been aimed at 1) solving the structure of proteins of interest related to the research projects of the groups. In some cases, the choice has been to select proteins of unknown function or of possible novel structure obtained from the sequenced genomes of the FAPESP genomic program; 2) providing the groups with training in all the steps of the protein structure determination: gene cloning, protein expression, protein purification, protein crystallization and structure determination. Having begun in 2001, the program has been successful in both aims. Here, four groups reveal their participation in the program and describe the structural aspects of the proteins they have selected to study. PMID- 16710563 TI - c-Ki-ras oncogene amplification and FGF2 signaling pathways in the mouse Y1 adrenocortical cell line. AB - The mouse Y1 adrenocortical tumor cell line is highly responsive to FGF2 (Fibroblast Growth Factor 2) and possesses amplified and over-expressed c-Ki-ras proto-oncogene. We previously reported that this genetic lesion leads to high constitutive levels of activation of the c-Ki-Ras-GTP-->PI3K-->Akt signaling pathway (Forti et al. 2002). On the other hand, activation levels of another important pathway downstream of c-Ki-Ras-GTP, namely, Raf-->MEK-->ERK, remain strictly dependent on FGF2 stimulation (Rocha et al. 2003). Here we show that, first, FGF2 transiently up-regulates the c-Ki-Ras-GTP-->PI3K-->Akt pathway, in spite of its high basal levels. Second, c-Ki-Ras-GTP transient up-regulation likely underlies activation of the ERK1/2 pathway by FGF2. Third, c-Ki-Ras-GTP high basal levels suppress activation of the c-H-Ras onco-protein. But, Y1 cells, expressing dominant negative mutant RasN17, display a rapid and transient up regulation of c-H-Ras-GTP upon FGF2 treatment. Elucidation of FGF2-signaling pathways in Y1 tumor cells can uncover new targets for drug development of interest in cancer therapy. PMID- 16710565 TI - Plasma membranes from insect midgut cells. AB - Plasma membranes from insect midgut cells are separated into apical and basolateral domains. The apical domain is usually modified into microvilli with a molecular structure similar to other animals. Nevertheless, the microvillar structure should differ in some insects to permit the traffic inside them of secretory vesicles that may budd laterally or pinch-off from the tips of microvilli. Other microvillar modifications are associated with proton-pumping or with the interplay with an ensheathing lipid membrane (the perimicrovilllar membrane) observed in the midgut cells of hemipterans (aphids and bugs). The perimicrovillar membranes are thought to be involved in amino acid absorption from diluted diets. The microvillar and perimicrovillar membranes have densities (and protein content) that depend on the insect taxon. The role played by the microvillar and perimicrovillar proteins in insect midgut physiology is reviewed here trying to provide a coherent picture of data and highlighting further research areas. PMID- 16710566 TI - Secretory organelles of pathogenic protozoa. AB - Secretory processes play an important role on the biology and life cycles of parasitic protozoa. This review focus on basic aspects, from a cell biology perspective, of the secretion of (a) micronemes, rhoptries and dense granules in members of the Apicomplexa group, where these organelles are involved in the process of protozoan penetration into the host cell, survival within the parasitophorous vacuole and subsequent egress from the host cell, (b) the Maurer's cleft in Plasmodium, a structure involved in the secretion of proteins synthesized by the intravacuolar parasite and transported through vesicles to the erythrocyte surface, (c) the secretion of macromolecules into the flagellar pocket of trypanosomatids, and (d) the secretion of proteins which make the cyst wall of Giardia and Entamoeba, with the formation of encystation vesicles. PMID- 16710567 TI - Sporothrix schenckii and sporotrichosis. AB - For a long time sporotrichosis has been regarded to have a low incidence in Brazil; however, recent studies demonstrate that not only the number of reported cases but also the incidence of more severe or atypical clinical forms of the disease are increasing. Recent data indicate that these more severe forms occur in about 10% of patients with confirmed diagnosis. The less frequent forms, mainly osteoarticular sporotrichosis, might be associated both with patient immunodepression and zoonotic transmission of the disease. The extracutaneous form and the atypical forms are a challenge to a newly developed serological test, introduced as an auxiliary tool for the diagnosis of unusual clinical forms of sporotrichosis. PMID- 16710568 TI - Garnets from the Camafuca-Camazambo kimberlite (Angola). AB - This work presents a geochemical study of a set of garnets, selected by their colors, from the Camafuca-Camazambo kimberlite, located on northeast Angola. Mantle-derived garnets were classified according to the scheme proposed by Grutter et al. (2004) and belong to the G1, G4, G9 and G10 groups. Both sub calcic (G10) and Ca-saturated (G9) garnets, typical, respectively, of harzburgites and lherzolites, were identified. The solubility limit of knorringite molecule in G10D garnets suggests they have crystallized at a minimum pressure of about 40 to 45 kbar (4-4.5 GPa). The occurrence of diamond stability field garnets (G10D) is a clear indicator of the potential of this kimberlite for diamond. The chemistry of the garnets suggests that the source for the kimberlite was a lherzolite that has suffered a partial melting that formed basaltic magma, leaving a harzburgite as a residue. PMID- 16710569 TI - Surface modification of a granite building stone in central Rio de Janeiro. AB - In order to evaluate environmental controls on the soiling formation and decay of building stones a set of mapping and physical and chemical analyses were carried out on granite from a historical church in the polluted centre of Rio de Janeiro. These techniques highlight the increasing of threatening damage on generally perceived as a durable building material, caused by granular disaggregation and contour scaling in areas close to ground level. Mapping also indicated the formation of black crusts over entire building facades, concentrated on areas sheltered from rain-wash. Analyses demonstrated the influence of marine aerosols, rock and mortar composition and mostly of the atmospheric pollutants on the decay and soiling of the granite. Much of the decay is associated specifically with the presence of halite (NaCl) and gypsum ((CaS04)2H2O). The fact that black, gypsum crusts are able to develop over entire facades in a humid subtropical environment is testimony to the high levels of local pollution, especially particulate deposition. Reduced rainwash, in sheltered micro-environments of narrow, canyon like streets, overcomes the gypsum tendency to be washed away from buildings facades. These observations further highlight that decay processes are primarily controlled by microclimatic conditions. PMID- 16710570 TI - Positive inversion of extensional footwalls in the southern Serra do Espinhaco, Brazil--insights from sandbox laboratory experiments. AB - Analogue experiments were carried out to get insights into the processes governing positive inversion during the foreland propagating thrust tectonics in the southern Serra do Espinhaco, a Brasiliano/Panafrican foldthrust belt in southeast Brazil. In particular, model listric half-grabens were inverted by applying contractional displacement to the footwall blocks. We investigated two different inversion conditions in listric half-grabens: (i) extensional and contractional detachments at the same level and (ii) at different positions. The models revealed that the development of a forward-breaking thrust system occurs in the basin synrift deposits, by contractional translation of the extensional footwall block when the extensional and contractional master faults do not coincide. Our experiments show the tectonic imbrication between basement and synrift sequences which characterizes the southern Serra do Espinhaco, and support the location in the eastern mountain range domain of the Espinhaco rift master fault system, which is not exposed at the surface. PMID- 16710571 TI - Contamination in mafic mineral-rich calc-alkaline granites: a geochemical and Sr Nd isotope study of the Neoproterozoic Piedade Granite, SE Brazil. AB - The Piedade Granite (approximately 600 Ma) was emplaced shortly after the main phase of granite magmatism in the Agudos Grandes batholith, Apiai-Guaxupe Terrane, SE Brazil. Its main units are: mafic mineral-rich porphyritic granites forming the border (peraluminous muscovite-biotite granodiorite-monzogranite MBmg unit) and core (metaluminous titanite-bearing biotite monzogranite BmgT unit) and felsic pink inequigranular granite (Bmg unit) between them. Bmg has high LaN/YbN (up to 100), Th/U (> 10) and low Rb, Nb and Ta, and can be a crustal melt derived from deep-seated sources with residual garnet and biotite. The core BmgT unit derived from oxidized magmas with high Mg# (approximately 45), Ba and Sr, fractionated REE patterns (LaN/YbN = 45), 87Sr/86Sr(t) approximately 0.710, epsilonNd(t) approximately -12 to -14, interpreted as being high-K calc-alkaline magmas contaminated with metasedimentary rocks that had upper-crust signature (high U, Cs, Ta). The mafic-rich peraluminous granites show a more evolved isotope signature (87Sr/86Sr(t) = 0.713-0.714; epsilonNd(t) = -14 to -16), similar to Bmg, and Mg# and incompatible trace-element concentrations intermediate between Bmg and BmgT. A model is presented in whichMBmgis envisaged as the product of contamination between a mafic mineral-rich magma consanguineous with BmgT and pure crustal melts akin to Bmg. PMID- 16710572 TI - Kinematics and geometry of structures in the southern limb of the Paraiba do Sul divergent structural fan, SE Brazil: a true transtensional shear. AB - Shear zones geometry in the Paraiba do Sul belt, southeastern Brazil, delineates a NE-trending fan-like structure. Shear zones dip towards SE in the northern limb, and towards NW in the southern one. This geometry has been interpreted either due to transpression or to late folding of flat-lying thrust surfaces. Stretching lineation plunges to ENE-ESE in the northern limb and towards NNE-NE in the southern one. Structural data in the southern limb of the divergent fan suggest a two stage kinematic evolution in high-temperature conditions: an earlier stage with top-to-SSW/SW sinistral thrusting and orogenic-parallel tangential motion, and a later stage with top-down to NNE/NE transtensional deformation. We propose a heterogeneous deformation model to explain the observed shear reversal, and suggest that the imposed transpressional displacement gradient may change during progressive deformation due to transient rheological inhomogeneities in bulk pure shear strain. In the earlier stage, the partially molten material could easily accommodate the imposed strain rates, giving rise firstly to the SW-directed shearing. As the thermal disturbance tended to vanish and the convergence increased, the NNE-directed transtensional shearing developed. We propose that the transtensional deformation characterized in this paper could be related to extrusion processes during regional transpressional strain. PMID- 16710573 TI - To follow or not to follow dermatological treatment--a review of the literature. AB - Creams, ointments and solutions applied to the skin surface by patients as part of a daily routine might be expected to provide a more variable dosage than do standard tablets. However, adherence to treatment in dermatology has been little studied. This article reviews recent publications in the field. These are dominated by questionnaire-based studies, which tend to over-estimate adherence. Reduced adherence to dermatological treatment is noted in 34-45% of patients. It is likely that the percentage of patients who practice truly optimal treatment in their daily life is even lower considering the variable practice of self treatment. Self-reported psychiatric morbidity contributes to poor adherence to dermatological treatment, while a well-functioning doctor-patient interaction is a major determinant of good adherence, as is patient satisfaction. In conclusion, adherence to dermatological treatment is unsatisfactory and there is a need for intervention and change in clinical routines. The therapeutic and economic benefits may be considerable. The immediate challenge is to stimulate a change in patient behaviour and improve self-treatment at home. PMID- 16710574 TI - Changes in quality of life in persons with eczema and psoriasis after treatment in departments of dermatology. AB - The objective of this study was to assess the impact of chronic dermatological diseases on quality of life (QoL) of Norwegian patients following in-patient management. QoL was measured by the Norwegian version of the Dermatology Life Quality Index, a validated, self-administered questionnaire. Adult in-patients with psoriasis and eczema were selected for one year from the dermatological departments in Norway. A total of 212 patients were included, and 126 patients (50% men, 85 with psoriasis and 41 with eczema, mean age 46 years) completed the questionnaires at time of hospital admission and one week after discharge. The patients reported adverse impact on QoL, but no differences between the psoriasis and eczema groups could be demonstrated. Patients with psoriasis improved from 18.3 (7.6) (mean (SD)) to 12.1 (8.2) (p < 0.01), and those with eczema improved from 20.0 (6.0) to 14.4 (7.8) (p < 0.01). Seventy percent of the patients showed improvement, 20% remained unchanged and 10% worsened. The parameters for which the most improvement was seen were those that were of most concern to the patients, i.e. their symptoms and embarrassment. In conclusion, the results are consistent with previous international studies. PMID- 16710575 TI - The role of dermatologists, nurses and pharmacists in chronic dermatological treatment: patient and provider views and experiences. AB - Effectively co-ordinated treatment support from healthcare providers (doctors, nurses and pharmacists) may improve patients' adherence to treatment. The objective of this study was to identify patients' and providers' perceptions of the roles of different healthcare providers in dermatological treatment. Focus groups were used in two types of fora: patients with chronic dermatological diseases (n =2x6) and healthcare providers (n =2x6), including doctors, nurses and pharmacists working in dermatological care. Data were analysed according to the Consensual Qualitative Research approach. The respondents viewed the roles of the providers as complementary, but poorly co-ordinated. Treatment support is provided mainly by the nurse. During the doctor's appointment, diagnosis and treatment decisions are often prioritized, leaving limited time for treatment support. The pharmacist's provision of support is constrained by the lack of privacy and clinical history of individual patients. The most apparent "gap" in the chain of treatment support was between the pharmacist and the other providers. There was a wish for improved interprofessional collaboration to avoid giving conflicting advice. There is a need to improve interprofessional collaboration in dermatology, in order to optimize treatment support in clinical practice. PMID- 16710576 TI - Autoimmune diseases and vitamin D receptor Apa-I polymorphism are associated with vitiligo in a small inbred Romanian community. AB - Vitiligo has been associated with the host's genetic profile, metabolic abnormality and immunostatus. The purpose of this study was to investigate the association of vitiligo with autoimmune diseases for 31 out of 39 subjects with vitiligo and their first-degree relatives living in a small Caucasian inbred rural community. They were compared with healthy individuals. A 2.28% prevalence of vitiligo was calculated and the presence of consanguine marriages (72.3%) was noted for this community. Our results indicate an increased prevalence of thyroidopathies, diabetes mellitus and rheumatoid arthritis in families with vitiligo. We also show that the Apa-I polymorphism of the vitamin D receptor gene is associated with vitiligo. This is the first study of its kind performed in Romania suggesting that the vitamin D receptor gene might play a role in the aetiopathogenesis of skin depigmentation. PMID- 16710577 TI - Diagnostic role of soluble fas ligand secretion by peripheral blood mononuclear cells from patients with previous drug-induced blistering disease: a pilot study. AB - Toxic epidermal necrolysis and Stevens-Johnson syndrome are severe blistering diseases generally considered to be hypersensitivity reaction to drugs. The aim of this study is to explore the diagnostic role of soluble Fas ligand secretion by peripheral blood mononuclear cells (PBMCs) in patients with previous drug induced blistering diseases with or without stimulation with the offending drug. The results revealed that PBMCs from the patient group showed significant soluble Fas ligand secretion after stimulation with 100 microg/ml culprit drug (p<0.05). No significant change was seen in the control groups (p>0.05). Taken as a group, PBMCs from the patient group secreted more soluble Fas ligand than the normal control group regardless of drug stimulation. These results indicate that individuals whose PBMCs are high secretors of soluble Fas ligand are prone to severe toxic epidermal necrolysis/Stevens-Johnson syndrome. The possibility of using soluble Fas ligand secretion as a tool for retrospective determination of culprit drugs in patients with previous drug-induced blistering disease awaits further investigation. PMID- 16710578 TI - Mortality and clinicopathological features of cutaneous squamous cell carcinoma in organ transplant recipients: a study of the Swedish cohort. AB - Solid organ transplant recipients have a high incidence of cutaneous squamous cell carcinoma and often develop multiple and aggressive tumours. This retrospective study based on the Swedish organ transplant cohort, focuses on the deaths caused by cutaneous squamous cell carcinoma and aims to elucidate the clinicopathological features of these tumours. The cohort comprised 5931 patients who underwent organ transplantation during the period 1970 to 1997 and were registered in the Swedish In-patient Registry, Cancer Registry and Causes-of Death Registry. A total of 544 cutaneous squamous cell carcinomas in 201 patients were re-examined. The dominating size of the tumours was 5-10 mm and one-third of the tumours were removed by methods other than excision surgery. Well differentiated tumours and Clark level IV were predominant. Seven patients died from their tumours, all of which were localized on the head. The principal site of metastasis was the parotid gland. The mean duration between date of transplantation and death was 10.4 years (range 6-17 years). Mortality from cutaneous cell carcinoma was compared with that of the general population. There was a highly increased risk; standardized mortality ratio 52.2; 95% confidence interval 21.0-107.6. However, the mortality rate in the Swedish cohort appears to be lower than what has been reported previously from other countries. PMID- 16710579 TI - 18S rDNA polymerase chain reaction and sequencing in onychomycosis diagnostics. AB - Diagnostic approaches to onychomycosis have traditionally been based on a combination of culture and microscopy. In the present study clinical specimens from 346 patients with suspected onychomycosis were analysed by 18S polymerase chain reaction (detection) followed by sequencing and subsequent database search (identification) in parallel with routine culture on agar (detection and identification). In 49 samples Trichophyton rubrum was identified by culture and sequencing. In 67 additional culture negative samples, a positive dermatophyte sequence was obtained (T. rubrum in 54, T. mentagrophytes in 5, and T. species in 8 samples). Fifteen samples cultured positive while no sequence was obtained. Two hundred and seven samples were negative by culture as well as by sequencing. Nails from 10 healthy controls were negative by culture and sequencing. In conclusion, the number of specimens that were positive by polymerase chain reaction was more than double the number that were positive by culture alone. PMID- 16710580 TI - A comparison of treatment of oral lichen planus with topical tacrolimus and triamcinolone acetonide ointment. AB - Treatment of symptomatic oral lichen planus remains a challenging problem. This study compared the efficacy of topical tacrolimus ointment with triamcinolone acetonide ointment in patients with oral lichen planus. Twenty patients (group I) were treated with topical tacrolimus 0.1% ointment 4 times daily, and 20 (group II) were treated with triamcinolone acetonide 0.1% ointment 4 times daily. The clinical effect was graded after 6 weeks. In group I, 6 patients healed, 12 showed improvement and 2 showed no improvement. In group II, 2 patients healed, 7 improved and 11 showed no improvement. The most commonly reported side-effect in both groups was temporary burning or stinging at the site of application. Unfortunately, oral lesions recurred within 3-9 weeks of cessation of treatment in 13 of the 18 patients who had initially shown an improvement or were healed in group I and in 7 of the 9 patients in group II. Topical tacrolimus 0.1% ointment induced a better initial therapeutic response than triamcinolone acetonide 0.1% ointment. However, relapses occurred frequently within 3-9 weeks of the cessation of treatment. PMID- 16710581 TI - Clonal spread of Staphylococcus aureus with reduced susceptibility to oxacillin in a dermatological hospital unit. AB - In November 2000, we became aware of isolates of Staphylococcus aureus with borderline resistance to oxacillin (BORSA) from patients in the Department of Dermatology, Aarhus University Hospital. The objective was to describe the isolates phenotypically and genotypically and to assess possible transmission routes in order to intervene and prevent further spread. Clonality of the isolates was confirmed by pulsed field gel electrophoresis. Several breaches in infection control procedures were revealed suggesting both direct and indirect transmission between patients. Defective skin barriers, high carrier rates of S. aureus in dermatological patients and high consumption rates of dicloxacillin in the department might facilitate transmission. Following improvement of the general infection control measures, and after reassessment of the antibiotic policy in the department, the outbreak has disappeared. PMID- 16710583 TI - Treatment of osmidrosis using the ultrasonic surgical aspirator. AB - Numerous surgical methods have been introduced to treat osmidrosis. However, many problems with wound healing have been reported, including haematoma, seroma, skin necrosis and severe scarring. Ultrasound energy liquefies sweat glands via cavitation, but minimally affects blood vessels and nerves at the same energy level. Our patient was treated with ultrasonic surgical aspiration, which allowed the effective removal of the apocrine glands with little scarring. The ultrasonic surgical aspirator offers a safe and effective method to treat osmidrosis with few complications. This is the operation of first choice, especially for the patients who are sensitive about their surgical scars. PMID- 16710582 TI - Ruby laser treatment of congenital melanocytic naevi--a pessimistic view. AB - Congenital melanocytic naevi may be disfiguring and potentially malignant. Treatment with lasers is a relatively new option, and promising results have been published. Most studies include few patients, and describe the effect of different lasers and different techniques. The aim of this report is to evaluate the results of ruby laser treatment on facial congenital naevi in children. Fourteen children (age range 2-16 years) with medium-sized congenital naevi were included. All naevi had a facial distribution, and were considered unsuitable for surgical excision. Ten children (age range 2-5 years) were treated under general anaesthesia and 4 under local anaesthesia. Q-switched and normal mode ruby lasers were used in 12 children and normal mode only in 2. Treatments were performed at intervals of 2-20 months (median 3 months), and the number of treatments varied between 2 and 8 (median 3). The results were monitored by clinical photographs. None of the patients showed a satisfactory result. Based on these results, laser treatment with ruby laser in children with congenital naevi is not recommended outside of clinical studies. Laser treatment may have a malignant potential that in our opinion is not balanced by good clinical results. PMID- 16710584 TI - Drug rash with eosinophilia and systemic symptoms induced by valproate and carbamazepine: formation of circulating auto-antibody against 190-kDa antigen. AB - Drug rash with eosinophilia and systemic symptoms (DRESS) is characterized by fever, rash and internal organ involvement after exposure to certain drugs. Most of the aromatic anticonvulsants, such as phenytoin, phenobarbital, and carbamazepine, can induce DRESS. Cross-sensitivity between not only the aromatic anticonvulsants, but also the aromatic and non-aromatic anticonvulsants, is possible. We report here a case of a Korean woman who had previously tolerated valproate, but developed DRESS due to valproate after carbamazepine intake, which indicates cross-sensitivity. Her skin lesions displayed diffuse oedematous patches on the entire body associated with tense bullae on her arms and legs. Circulating auto-antibody to 190-kDa antigen was detected in the patient's serum by indirect immunofluorescence and immunoblotting, which might contribute to a pathogenic role in DRESS. PMID- 16710586 TI - Persistent post-occupational dermatitis: report of five cases. AB - Persistent post-occupational dermatitis is a phenomenon that is well-recognized by occupational dermatologists, but there have been few studies on it. In view of this, we proposed to assess the prevalence of this phenomenon in an English setting and ascertain the characteristics of the patients affected. Using modified criteria adapted from previous studies, details of 1100 patients seen in a contact dermatitis clinic were screened. Persistent post-occupational dermatitis was diagnosed in 5 patients out of 1100 seen over a 35-month period in a contact clinic (4 women, 1 man; age of onset 19-52 years). All had hand dermatitis that persisted despite removal of the apparent causative agents. Four patients were nickel-allergic on patch testing, though nickel was thought to be a potential causative agent in only one case and 2 patients were allergic to thiuram-mix on patch testing, and in both thiuram had a possible causative role. In all 5 cases, irritant exposure seemed important, with allergic factors contributing in 3 cases. Two patients had had eczema in childhood. Persistent post-occupational dermatitis is uncommon, affecting less than 0.5% of patients seen in a contact dermatitis clinic, but when diagnosed it has major implications for the future employment prospects of the individuals concerned. PMID- 16710585 TI - A fatal case of malignant atrophic papulosis (Degos' disease) in a man with factor V Leinden mutation and lupus anticoagulant. AB - Malignant atrophic papulosis (Degos' disease) is a very rare condition characterized by atrophic papular skin lesions and variable association of systemic involvement. We describe a 33-year-old man who presented with a widespread skin eruption consistent with malignant atrophic papulosis. During the course of the disease he even developed penile ulcerations, a symptom that has been reported only a few times previously. He subsequently died of multiple perforations of the small bowel 2.5 years after onset of the disease. Laboratory investigations revealed a mutation of factor V Leiden and the presence of lupus anticoagulant, but no anti-cardiolipin antibodies. The patient was treated with narrow-band ultraviolet (UV)B, prednisolone and, later, aspirin, pentoxifyllin and warfarin. Despite this very intensive anticoagulant and anti-platelet therapy, the treatment had no effect on the skin lesions and could not prevent systemic involvement. PMID- 16710587 TI - Scoring of skin lesions with the perioral dermatitis severity index (PODSI). PMID- 16710588 TI - Oesophageal involvement in familial benign chronic pemphigus. PMID- 16710589 TI - Treatment of recalcitrant dermatomyositis with efalizumab. PMID- 16710590 TI - Severe acne successfully treated with etanercept. PMID- 16710591 TI - Rosacea fulminans related to pegylated interferon alpha-2b and ribavirin therapy. PMID- 16710593 TI - Subungual basal cell carcinoma in an elderly Japanese woman. PMID- 16710592 TI - Vegetating darier's disease during pregnancy. PMID- 16710594 TI - Early diagnosed but late treated subungual melanoma. PMID- 16710595 TI - Morpheaform sarcoidosis. PMID- 16710597 TI - Scalp epithelioid glomus tumor: a rare location. PMID- 16710596 TI - A neurofibroma with unusual morphology. PMID- 16710598 TI - Primary cutaneous blastic natural killer-cell lymphoma. PMID- 16710599 TI - Multiple haemangiomas in a psoriatic arthritis patient treated with cyclosporine. PMID- 16710600 TI - Hepatocellular focal nodular hyperplasia after danazol treatment for hereditary angio-oedema. PMID- 16710601 TI - Extracutaneous tumour mass following bexarotene and interferon therapy of primary cutaneous CD30+ large cell lymphoma. PMID- 16710602 TI - Enhanced expression of p53 protein in pityriasis rubra pilaris. PMID- 16710603 TI - An umbilical nodule due to endometriosis. PMID- 16710604 TI - Haemochromatosis gene mutations and response to chloroquine in sporadic porphyria cutanea tarda. PMID- 16710605 TI - Addition of a topical corticosteroid in the early phase of alefacept treatment for psoriasis. PMID- 16710606 TI - Necrotizing livedo related to neoplastic microvascular embolism. PMID- 16710607 TI - Dimensionality of the dermatology life quality index (DLQI): a commentary. PMID- 16710608 TI - Human beta-defensins. AB - The last decade led to the discovery and characterization of several human beta defensins. Analysis of genomic information indicates that the number of beta defensin-like molecules encoded by the human genome may number in the tens. Growing interest in beta-defensins steadily enhances our knowledge about various aspects of their gene location, expression patterns and the transcription factors involved in their regulation in vivo. The hallmark property of beta-defensins, their antimicrobial activity, is clearly only the tip of the iceberg in the extensive network of inter-relations within the immune system in which these peptides function. Structural studies of beta-defensins provide the molecular basis for a better understanding of their properties, functions and their potential for practical applications. In this review, we present some recent advances in the studies of human beta-defensins, with an emphasis on possible correlations between their structural and functional properties. PMID- 16710609 TI - Role of dystrophin and utrophin for assembly and function of the dystrophin glycoprotein complex in non-muscle tissue. AB - The dystrophin glycoprotein complex (DGC) is a multimeric protein assembly associated with either the X-linked cytoskeletal protein dystrophin or its autosomal homologue utrophin. In striated muscle cells, the DGC links the extracellular matrix to the actin cytoskeleton and mediates three major functions: structural stability of the plasma membrane, ion homeostasis, and transmembrane signaling. Mutations affecting the DGC underlie major forms of congenital muscle dystrophies. The DGC is prominent also in the central and peripheral nervous system and in tissues with a secretory function or which form barriers between functional compartments, such as the blood-brain barrier, choroid plexus, or kidney. A considerable molecular heterogeneity arises from cell-specific expression of its constituent proteins, notably short C-terminal isoforms of dystrophin. Experimentally, the generation of mice carrying targeted gene deletions affecting the DGC has clarified the interdependence of DGC proteins for assembly of the complex and revealed its importance for brain development and regulation of the 'milieu interieur. Here, we focus on recent studies of the DGC in brain, blood-brain barrier and choroid plexus, retina, and kidney and discuss the role of dystrophin isoforms and utrophin for assembly of the complex in these tissues. PMID- 16710614 TI - Hemoglobin-based oxygen carriers in trauma care: scientific rationale for the US multicenter prehosptial trial. AB - BACKGROUND: The greatest need for blood substitutes worldwide is in patients with unanticipated acute blood loss, and trauma is the most likely scenario. The blood substitutes reaching advanced clinical trials today are red blood cell (RBC) substitutes derived from hemoglobin. The hemoglobin-based oxygen carriers (HBOCs) tested currently in advanced clinical trials are polymerized hemoglobin solutions. METHODS: In the USA, the standard approach to restoring oxygen delivery for hemorrhagic shock has been crystalloid administration to expand intravascular volume, followed by stored RBCs for critical anemia. Allogeneic RBCs, however, may have adverse immunoinflammatory effects that increase the risk of postinjury multiple organ failure (MOF). Phase II in hospital clinical trials, as well as in vitro and in vivo work, suggest that resuscitation with an HBOC--in lieu of stored RBCs--attenuates the systemic inflammatory response invoked in the pathogenesis of MOF. Specifically, an HBOC has been shown to obviate stored RBC provoked polymorphonuclear neutrophil (PMN) priming, endothelial activation, and systemic release of interleukins (IL) 6, 8, and 10. In a 2-event rodent study of shock-induced PMN-mediated acute respiratory distress syndrome (ARDS), the simulated prehospital administration of an HBOC markedly attenuated lung injury. RESULTS: Based on this background and work by others, we have initiated a US multicenter prehospital trial in which severely injured patients with major blood loss [systolic blood pressure (SBP)500 injured patients have been enrolled in this multicenter trial, and the final interim analyses support the original target of 720. PMID- 16710628 TI - Incarceration of umbilical hernia during pregnancy due to a sessile fibroid. AB - The incidence of both umbilical hernias and fibroids during pregnancy is reported to be rare. Another rare entity is the incarceration of fibroids in pregnancy. We report here the case of a 30-year-old woman in her 32nd gestational week with an incarcerated umbilical hernia. She was operated on an emergency basis, the sessile fibroid was mobilized and the hernia defect was repaired with the Mayo technique. No complications were observed during the post-operative period. She gave birth to a baby girl during the 38th gestational week by means of a cesarean section. The possibility of a fibroid entrapped in the hernial sac should always be taken into consideration at the evaluation of incarcerated hernias during pregnancy. If the contents of the hernial sac cannot be pushed into the abdominal cavity easily, an emergency operation should be considered as a means to prevent further complications related to pregnancy. PMID- 16710627 TI - Pain and functional impairment 6 years after inguinal herniorrhaphy. AB - Pain impairing daily activities following inguinal herniorrhaphy is reported by about 10% of patients, when asked 1-2 years postoperatively. However, the time course and consequences of postherniorrhaphy pain is not known in detail. A nationwide follow-up questionnaire study was undertaken 6.5 years postoperatively in 335 well-described patients reporting pain 1 year after inguinal herniorrhaphy in a previous questionnaire study. Three hundred and three patients, who were alive and could be contacted, received a questionnaire 6.5 years after the herniorrhaphy. Response rate was 88%. Of 267 patients responding, 57 were analyzed separately due to subsequent inguinal herniorrhaphy or other major surgery in the observation period, leaving 210 patients (69.3%) for primary analysis. Pain from the previous hernia site was reported by 72 patients (34.3%), and 52 patients (24.8%) reported that pain affected daily activities. Less pain, compared to the 1-year follow-up, was reported by 75.8%, while 16.7% had the same intensity level and 7.5% reported increased pain severity. In the subgroup of patients operated for a recurrence during the observation period and not included in primary analysis, 22 of 44 (50%) still experienced pain at 6.5 years, and 17 (38.6%) reported that pain affected daily activities (mean observation period 4.5 years). Pain after inguinal herniorrhaphy decreased from about 11% 1 year after surgery, but still affects daily activities in about 6% after 6.5 years. Patients operated for a recurrence are at higher risk for persistent pain. PMID- 16710631 TI - E7 proteins from high- and low-risk human papillomaviruses bind to TGF-beta regulated Smad proteins and inhibit their transcriptional activity. AB - Human papillomaviruses (HPV) infect keratinocytes of skin and mucosa. Persistent infection can lead to the formation of benign tumors. In cases of high-risk HPV, such as HPV16 or 18, these may further progress to cancer. In order to support viral replication in suprabasal keratinocytes, the HPV E7 protein employs various strategies to keep keratinocytes in cycle and counteracts anti-proliferative signals from outside. HPV16 E7 can directly interfere with transforming growth factor-beta (TGF-beta) signalling by binding to Smad proteins mediating growth arrest. It has been speculated that this property of HPV16 E7 contributes to HPV associated carcinogenesis. Here, we show that E7 proteins from different low- and high-risk HPV types bind to Smad 1 to 4. The E7 protein from HPV1, a low-risk HPV causing plantar warts, efficiently inhibited Smad 3-induced transcription. Our data strongly indicate that the Smad-binding capacity of E7 proteins from different HPVs may preserve keratinocyte proliferation required for the productive viral life cycle rather than promoting carcinogenesis. PMID- 16710632 TI - Bone grafts impregnated with antibiotics as a tool for treating infected implants in orthopedic surgery - one stage revision results. AB - Infection of an orthopedic implant is considered a devastating complication, necessitating its complete removal and thorough debridement of the site. Osseous defects are common in such conditions and need to be addressed before a new implant may be inserted. So far bone grafting has been contraindicated in bacterially contaminated areas and could only be performed as soon as all signs of infection have ceased. Usually long term antibiotic treatment and a multitude of surgical interventions within a period of several months is required until a definitive supply can be achieved. Allograft bone may be impregnated with high loads of antibiotics using special incubation techniques. Based on this technology 48 exchange procedures of infected orthopaedic implants were performed in a single stage, all of them without the use of bone cement. There were 37 infected hips, 8 knees and 3 infected osteosyntheses. Two hips required re revision because of persisting infection, the remaining 46 patients stayed infect free for a period between 1 and 7 years after surgery. No adverse side effects could be found. Incorporation appeared as after grafting with unimpregnated bone grafts. Antibiotic loaded allograft bone is a powerful tool in septic revision surgery, enabling restoration of bone stock, insertion of a new implant and control of infection in a single operation. PMID- 16710633 TI - Salt-activated endoglucanase of a strain of alkaliphilic Bacillus agaradhaerens. AB - An endoglucanase was purified to homogeneity from an alkaline culture broth of a strain isolated from seawater and identified here as Bacillus agaradhaerens JAM KU023. The molecular mass was around 38-kDa and the N-terminal 19 amino acids of the purified enzyme exhibited 100% sequence identity to Cel5A of B. agaradhaerens DSM8721(T). The enzyme activity increased around 4-fold by the addition of 0.2 2.0 M NaCl in 0.1 M glycine-NaOH buffer (pH 9.0). KCl, Na2SO4, NaBr, NaNO3, CH3COONa, LiCl, NH4NO3, and NH4Cl also activated the enzyme up to 2- to 4-fold. The optimal pH and temperature values were pH 7-9.4 and 60 degrees C with 0.2 M NaCl, but pH 6.5-7 and 50 degrees C without NaCl; enzyme activity increased approximately 6-fold at 60 degrees C with 0.2 M NaCl compared to that at 50 degrees C without NaCl in 0.1 M glycine-NaOH buffer (pH 9.0). The thermostability and pH stability of the enzyme were not affected by NaCl. The enzyme was very stable to several chemical compounds, surfactants and metal ions (except for Fe2+ and Hg2+ ions), regardless whether NaCl was present or not. PMID- 16710634 TI - Global transcriptomic analysis of Desulfovibrio vulgaris on different electron donors. AB - Whole-genome microarrays of Desulfovibrio vulgaris were used to determine relative transcript levels in cells grown to exponential or stationary phase on a medium containing either lactate or formate as electron donor. The results showed that 158 and 477 genes were differentially expressed when comparing exponential to stationary phase in lactate- or formate-based media, respectively; and 505 and 355 genes were responsive to the electron donor used at exponential or stationary phase, respectively. Functional analyses suggested that the differentially regulated genes were involved in almost every aspect of cellular metabolism, with genes involved in protein synthesis, carbon, and energy metabolism being the most regulated. The results suggested that HynBA-1 might function as a primary periplasmic hydrogenase responsible for oxidation of H2 linked to the proton gradient in lactate-based medium, while several periplasmic hydrogenases including HynBA-1 and Hyd might carry out this role in formate-based medium. The results also indicated that the alcohol dehydrogenase and heterodisulfide reductase catalyzed pathway for proton gradient formation might be actively functioning for ATP synthesis in D. vulgaris. In addition, hierarchical clustering analysis using expression data across different electron donors and growth phases allowed the identification of the common electron donor independent changes in gene expression specifically associated with the exponential to stationary phase transition, and those specifically associated with the different electron donors independent of growth phase. The study provides the first global description and functional interpretation of transcriptomic response to growth phase and electron donor in D. vulgaris. PMID- 16710635 TI - Occurrence and pathogenic potential of Bacillus cereus group bacteria in a sandy loam. AB - The major part (94%) of the Bacillus cereus-like isolates from a Danish sandy loam are psychrotolerant Bacillus weihenstephanensis according to their ability to grow at temperatures below 7 degrees C and/or two PCR-based methods, while the remaining 6% are B. cereus. The Bacillus mycoides-like isolates could also be divided into psychrotolerant and mesophilic isolates. The psychrotolerant isolates of B. mycoides could be discriminated from the mesophilic by the two PCR based methods used to characterize B. weihenstephanensis. It is likely that the mesophilic B. mycoides strains are synonymous with Bacillus pseudomycoides, while psychrotolerant B. weihenstephanensis, like B. mycoides, are B. mycoides senso stricto. B. cereus is known to produce a number of factors, which are involved in its ability to cause gastrointestinal and somatic diseases. All the B. cereus like and B. mycoides like isolates from the sandy loam were investigated by PCR for the presence of 12 genes encoding toxins. Genes for the enterotoxins (hemolysin BL and nonhemolytic enterotoxin) and the two of the enzymes (cereolysin AB) were present in the major part of the isolates, while genes for phospolipase C and hemolysin III were present in fewer isolates, especially among B. mycoides like isolates. Genes for cytotoxin K and the hemolysin II were only present in isolates affiliated to B. cereus. Most of the mesophilic B. mycoides isolates did not possess the genes for the nonhemolytic enterotoxin and the cereolysin AB. The presence of multiple genes coding for virulence factors in all the isolates from the B. cereus group suggests that all the isolates from the sandy loam are potential pathogens. PMID- 16710636 TI - Identification and characterization of a germination operon from Bacillus thuringiensis. AB - Spore cortex-lytic enzymes are essential for germination in Bacillus. A homologue of the cwlJ gene involved in spore germination was isolated from Bacillus thuringiensis. The deduced product of this gene exhibits striking sequence similarity to CwlJ of Bacillus subtilis. Another open reading frame (ORF) was found 27 bp downstream of cwlJ and its deduced product shows high similarity to YwdL of B. subtilis. Reverse transcription polymerase chain reaction analysis indicated that cwlJ and ywdL formed a bicistronic operon in B. thuringiensis. Disruption of this operon did not affect sporulation and the spores of corresponding mutant showed the same refractility as the wild-type strain. In contrast, the fall of optical density at 600 nm in the mutant culture was slower than that of the wild-type strain during the spore germination response to different germinants such as L: -alanine, inosine or CaDPA. The spore germination of the cwlJ mutant was restored by introducing this operon into the disruption mutant. These results suggest this operon is essential for normal spore germination in B. thuringiensis. The expression of cwlJ was observed in the sporulating cells through Western blot experiments. PMID- 16710637 TI - Bensingtonia pseudonaganoensis sp. nov., a novel ballistoconidium-forming yeast species isolated from plant leaves. AB - Among the basidiomycetous yeasts isolated from plant leaves collected in different regions of China, two ballistoconidium-forming strains were revealed to represent an undescribed species of the genus Bensingtonia by conventional, chemotaxonomic and molecular phylogenetic characterization. Sequence analysis of the 26S rDNA D1/D2 domains and the internal transcribed spacer (ITS) region indicated that the novel species was located in the Agaricostilbum lineage and closely related to Bensingtonia naganoensis and Bensingtonia ciliata, with the former as its closest relative. The name Bensingtonia pseudonaganoensis sp. nov. is proposed (type strain: AS 2.2601T = CBS 10121T). PMID- 16710638 TI - Quorum-sensing system influences root colonization and biological control ability in Pseudomonas fluorescens 2P24. AB - Pseudomonas fluorescens 2P24 is a biocontrol agent isolated from a wheat take-all decline soil in China. This strain produces several antifungal compounds, such as 2,4-diacetylphloroglucinol (2,4-DAPG), hydrogen cyanide and siderophore(s). Our recent work revealed that strain 2P24 employs a quorum-sensing system to regulate its biocontrol activity. In this study, we identified a quorum-sensing system consisting of PcoR and PcoI of the LuxR-LuxI family from strain 2P24. Deletion of pcoI from 2P24 abolishes the production of the quorum-sensing signals, but does not detectably affect the production of antifungal metabolites. However, the mutant is significantly defective in biofilm formation, colonization on wheat rhizosphere and biocontrol ability against wheat take-all, whilst complementation of pcoI restores the biocontrol activity to the wild-type level. Our data indicate that quorum sensing is involved in regulation of biocontrol activity in P. fluorescens 2P24. PMID- 16710640 TI - Variation between freshwater and terrestrial fungal communities on decaying bamboo culms. AB - Fungal communities on decaying culms of a bamboo host (Phyllostachys bambusoides) from freshwater and adjacent terrestrial habitats were identified. Collections were made at Xiao Bai Long Mountain, Yiliang, Yunnan, China in the winter and summer. In each collection, 100 similar-sized bamboo culms were collected, comprising 50 submerged samples from a stream and 50 terrestrial samples from adjacent riparian vegetation. A total of 82 fungal taxa were recorded from the samples, including 30 ascomycetes and 52 anamorphic fungi. The frequency of occurrence of these fungi were recorded and the Shannon-Weiner indices (H') were applied to evaluate fungal diversity. The results showed that variation of the fungal diversity between the summer and winter collections was insignificant (0.2 < p < 0.5). Fungal diversity on submerged bamboo however, was significantly higher than that on terrestrial bamboo (p < 0.001). Further findings were that: (1) some commonly recorded freshwater and terrestrial taxa were found in both habitats, but overall there were only 15 overlapping species between the two habitats; (2) the dominant species in each habitat were considerably different, and (3) only a few fungi were dominant, while most species were rare, being recorded only once or twice. Factors responsible for the distribution patterns and variations in composition of the fungal communities are discussed. PMID- 16710639 TI - Isolation of mutations synthetic-lethal to prohibitin 2 null mutants of Saccharomyces cerevisiae. AB - Prohibitins are ubiquitous, abundant proteins found in a wide range of organisms and that have a high degree of sequence conservation. In yeast it has previously been demonstrated that prohibitin proteins form a complex and are involved in maintaining the morphological and functional integrity of mitochondria. We have used a colony-sectoring assay as a screen for mutants that are dependent upon the presence of functional Phb2p in the cell. Two classes of prohibitin dependent mutation (pbd1 and pbd2) were isolated and characterised. The effect of these mutations on replicative lifespan was determined, demonstrating that the pbd1 mutant slightly extended lifespan, whereas in contrast, the pbd2 mutation resulted in a shortening in both the mean- and the maximum-lifespan. The pbd1 mutation was also found to reduce chronological lifespan. Reducing the expression of the PHB2 gene in the pbd mutants was found to retard the rate of growth and to affect replicative lifespan. As the two mutants behave in a different manner they probably affect different aspects of prohibitin function. PMID- 16710641 TI - Abstracts of the Sixteenth Meeting of the European Neurological Society. May 27 31, 2006. Lausanne, Switzerland. PMID- 16710642 TI - The association of serum total sialic acid/total protein ratio with diabetic parameters in young type 1 diabetic patients. AB - Sialic acid is a terminal component of the non-reducing end of carbohydrate chains of glycoproteins and glycolipids. The purpose of this study was to estimate serum total sialic acid (TSA) concentrations and serum TSA/serum total protein (TP) ratios in young type 1 diabetic subjects and to investigate their association with diabetes-related parameters in that population. Twentyfour young type 1 diabetic patients and 20 healthy controls were enrolled in this study. Serum TSA and serum TSA/TP ratio were measured in both groups. Moreover, we looked for correlation among serum TSA, serum TSA/TP ratio and clinically relevant parameters such as urinary albumin excretion, blood pressure, diabetes duration, HbA1c, daily insulin dose, serum lipids and magnesium in type 1 diabetic patients. Serum TSA concentrations and serum TSA/TP ratio showed no statistical difference between patients and controls (p>0.05). While serum TSA concentrations only correlated with urinary albumin excretion (r=0.44, p=0.028), serum TSA/TP ratio correlated with diastolic blood pressure (r=0.48, p=0.015), diabetes duration (r=0.46, p=0.022) and urinary albumin excretion (r=0.53, p=0.007) in the diabetic subjects. We concluded that serum TSA/TP ratio might be a better indicator than serum TSA as an index of diabetic complications. PMID- 16710643 TI - Safety profile of repaglinide as used in general practice in England: results of a prescription-event monitoring study. AB - Repaglinide is a prandial glucose regulator indicated for management of type 2 diabetes. This post-marketing study used the observational cohort technique of prescription-event monitoring (PEM) to monitor safety of repaglinide prescribed in primary care in England. Patients were identified from dispensed prescriptions issued by general practitioners (GPs) between December 1998 and January 2001. Demographic and clinical event data were collected from questionnaires posted to GPs at least six months after the date of first prescription for each patient. The cohort consisted of 5731 patients [median age 60 (IQR 51-68), 49.9% male]. Event incidence densities (IDs) [no. 1st reports/1000 patient-months of exposure] were calculated for all events reported. The most frequently recorded clinical events in the first month were diarrhoea (ID(1) 10.3), malaise/lassitude (ID(1) 8.1) and nausea/vomiting (ID(1) 7.9). The most frequently reported reason for stopping was 'not effective' (647), with the most common clinical reasons being diarrhoea (60), malaise/lassitude (55) and intolerance (54). One hundred and thirteen adverse drug reactions (ADRs) were reported, with the most frequently specified being diarrhoea (10), abdominal pain (10) and nausea/vomiting (9). We concluded that repaglinide is generally well tolerated when used in general practice in England and did not identify any serious unrecognised adverse events. PMID- 16710644 TI - Thyroid dysfunction and thyroid autoimmunity in Saudi type 2 diabetics. AB - Diabetes mellitus and thyroid disease are common endocrine disorders in the general population. To investigate the association between thyroid dysfunction, thyroid autoimmunity and Saudi type 2 diabetics, a random sample of 100 Saudi type 2 diabetics and 100 age- and sex-matched controls were studied. The mean age was 54 years for diabetics and 55 years for controls while the male:female ratios were 1:1.6 and 1:14 respectively. GAD65ab were found in 26% diabetics and 2% controls (p=0.001). Thyroid autoimmunity were detected in 10% diabetics vs. 5% controls (p=0.05), while thyroid dysfunction was found in 16% and 7% respectively (p=0.03). In GAD65ab-positive diabetics, thyroid autoimmunity was observed in 27% vs. 4% GAD65ab-negative diabetics (p=0.02) and thyroid dysfunction was reported in 42% and 7% respectively. We conclude that thyroid dysfunction and autoimmunity are common in Saudi type 2 diabetics. Further studies are needed on the cost effectiveness of thyroid screening in diabetics. PMID- 16710645 TI - Simultaneous, painless, homolateral oculomotor and trochlear nerve palsies in a patient with type 2 diabetes mellitus. Neuropathy or brainstem infarction? AB - Combined palsies of cranial nerves, especially of the oculomotor nerves, are distinctly uncommon, even in patients with diabetes mellitus. We present a patient with type 2 diabetes mellitus, arterial hypertension and hyperlipidaemia, who had simultaneous oculomotor and trochlear nerve palsies. An MRI scan showed multiple brainstem ischaemic infarcts. The patient was treated with intensified insulin regimen and clopidogrel. Symptoms gradually improved, and at 9 months there was no further improvement. PMID- 16710646 TI - How to start or change a daily insulin regimen without a specialist in diabetes. Effectiveness and RAND assessment of practical schemes for in-hospital or hospital-based home care services use. AB - When the diabetic patient is hospitalised or assisted by a hospital-based home care service, as a rule he/she is not cared for by a diabetologist all day long. The aim of the present work was to perform a RAND assessment of practical schemes to start or change a daily insulin regimen without a diabetologist. We created an expert panel of seven internists and two diabetologists. They judge the appropriateness of each practical scheme using the RAND method. We produced 21 clinical scenarios divided into two groups. The first group of 9 clinical scenarios-i.e., for diabetic patients who 'eat/not-always-eat' and are 'normal weight/obese/underweight' or with 'renal failure' or 'defedated/end-of-life' or 'vomiting' or on 'parenteral nutrition'-useful for starting a daily insulin regimen without a diabetologist had a median RAND score of 8 (range 7-9). The second one-formed by 12 clinical scenarios useful to change the daily insulin dosage without a diabetologist based on low or high capillary glucose level monitoring-had a median RAND score of 9 (range 7-9). There was a high level of agreement between panellists in judging 'appropriate' the practical schemes to start or change a daily insulin regimen without a diabetologist. PMID- 16710647 TI - The restorative effects of pulsed infrared light therapy on significant loss of peripheral protective sensation in patients with long-term type 1 and type 2 diabetes mellitus. AB - Pulsed infrared light therapy (PILT) has been shown to increase peripheral sensation in diabetic patients with diabetic peripheral neuropathy (DPN). However, most studies last for very short periods, with the subjects receiving only 6-20 treatments. The purpose of this study was to evaluate the effectiveness of an eight-week course of PILT in reversing long-standing, profound DPN in patients with type 1 and type 2 diabetes. Twenty-two subjects with a diagnosis of type 1 (n=2) or type 2 (n=20) diabetes participated in the study. PILT was administered to one foot chosen at random with the other foot serving as a within subject control (no treatment). Patients underwent 24 treatments (3 times/week, for eight weeks) for 30 min per treatment. Changes in peripheral protective sensation (PPS) were measured using Semmes-Weinstein monofilaments (SWM) ranging from 3.7 to 6.48. PILT improved PPS even in patients with long-standing chronic neuropathies whose initial pre-study sensation was not measurable with a 200-g SWM. PILT significantly improves PPS. While the exact mechanism of action is not understood, infrared light may improve peripheral neuropathies by improving foot perfusion by stimulating nitric oxide production. PMID- 16710648 TI - Large decreases in insulin requirement occurred repeatedly in two pregnancies in a type 1 diabetic woman. AB - Whereas the increase in insulin requirement in late gestation is well described in diabetic pregnancy, a decrease in insulin dosage in the end of the third trimester has also been noted in clinical experience and the prevention of hypoglycaemia is important. Here we report a large decrease (over 50%) in the final few weeks of her two gestations in a woman with type 1 diabetes mellitus. PMID- 16710650 TI - Where do we belong? Urban adolescents' struggle for place and voice. AB - Through a telling of key events in the history of the "Teen Action Center" (TAC), a drop-in youth center located in downtown "Unionville," this story demonstrates how 'youth' is an important diversity category. The community conflict highlighted in this story centers around the 1997 arrest of TAC's Executive Director and two youth leaders (all Puerto Rican) because a small group of Latino and African-American youth was smoking cigarettes on the sidewalk in front of the Center. This conflict brings into focus divergent views on where Unionville's youth of color belong in the city, both physically and figuratively. The lessons learned in this story have wide application as Unionville, and other cities undergoing demographic transformation and economic decline, are likely to continue to experience these types of clashes, where the dominant paradigm of economic development overrides the realities, rights, and interests of marginalized groups. PMID- 16710651 TI - [Structural quality of rheumatology clinics for children and adolescents. Paper by a task force of the "Society of Pediatric and Adolescent Rheumatology" and of the "Association of Rheumatology Clinics in Germany"]. AB - Rheumatic diseases in childhood and adolescence differ from those of adulthood according to type, manifestation, treatment and course. A specialized therapy, starting as early as possible, improves the prognosis, can prevent long-term damage and saves the costs of long-term care. Only a specialized pediatric care system can guarantee optimum quality of the processes involved and the results for rheumatology in childhood and adolescence within a global financial system. This requires adequate structural quality of the specialized clinics and departments for pediatric rheumatology. The management of rheumatic diseases in childhood and adolescence is comprehensive and requires a multidisciplinary, specialized and engaged team which can cover the whole spectrum of rheumatic diseases with their various age-dependent aspects. In order to guarantee an adequate, cost-efficient routine, a specialized center which concentrates on inpatient care should treat at least 300 patients with pediatric rheumatic diseases per year. The diagnoses should be divided among the various disease categories with at least 70% of them involving inflammatory rheumatic diseases. For the inpatient care of small children, an accompanying person (parent) is necessary, requiring adequate structures and services. Patient rooms as well as diagnostic (radiography, sonography, etc.) and therapeutic services (physiotherapy, occupational therapy, pool, etc.) must be adequate for small children and school children as well as adolescents. Suitable mother-child units must also be provided and a school for patients is required within the clinic. A pediatric rheumatologist must be available 24 h a day, and it must be possible to reach other specialists within a short time. For painful therapeutic procedures, age-appropriate pain management is obligatory. A continuous adjustment of these recommendations to changing conditions in health politics is intended. PMID- 16710652 TI - The relationship between function, quality of life and coping in patients with low-grade gliomas. AB - OBJECTIVES: The principal aim of the study was to describe function, quality of life and coping with illness-related problems in patients with low-grade gliomas (LGG), to evaluate the need of support. A second aim was to investigate how function, quality of life and coping were related. MATERIALS AND METHODS: Thirty nine patients with a diagnosis of LGG answered the European Organization for Research and Treatment of Cancer Core Quality of Life Questionnaire (EORTC-QLQ C30) and the Ways of Coping Questionnaire (WCQ). The patients' level of function was assessed in accordance with the WHO performance status scale. RESULTS AND DISCUSSION: Nearly all patients were capable of self-care, but less than half were able to carry out normal activities without restriction. Problems with fatigue, sleep disturbances and pain were most frequent. Most difficulties were reported in the domains, Role, Cognitive and Emotional functioning. Seventeen patients (45%) had scores indicating low overall quality of life. Ratings of overall quality of life and fatigue had the strongest relationship. The trend in the results suggested that mental problems have a stronger impact on quality of life than physical ones. Emotion-focused coping dominated. There was a significant relationship between coping by escape-avoidance and lower level of emotional functioning. CONCLUSIONS: The results of this study indicate that a multidisciplinary team is needed for assessment and treatment of the different problems in patients with LGG. Furthermore, special attention should be paid to patients who use coping by escape-avoidance, as this is associated with emotional distress. PMID- 16710657 TI - Putaminal involvement in Rasmussen encephalitis. AB - BACKGROUND: Rasmussen encephalitis (RE) is a rare devastating disease of childhood causing progressive neurological deficits and intractable seizures, typically affecting one hemisphere. Characteristic MRI features include progressive unihemispheric focal cortical atrophy and grey- or white-matter high signal changes and basal ganglion involvement, particularly of the caudate nucleus. OBJECTIVE: To analyse the pattern of involvement of different brain structures in a series of patients with RE and to attempt clinical correlation. MATERIALS AND METHODS: We reviewed the medical records and neuroimaging data of 12 patients diagnosed with RE satisfying the European Consensus Statement diagnostic criteria. RESULTS: The disease manifested as seizures in all patients and was refractory; epilepsia partialis continua was a notable feature (nine patients). Hemiparesis of varying grades was noted in all but one patient; none had extrapyramidal signs. Neuroimaging showed cortical involvement in the insular/periinsular regions in 11 patients. Caudate atrophy was noted in ten patients. Putaminal atrophy was seen in nine patients, six of whom had additional hyperintense signal changes. CONCLUSIONS: Our study highlights frequent putaminal atrophy and signal changes in RE, which suggests a more extensive basal ganglion involvement than emphasized previously. Recognition of putaminal changes may be a useful additional tool in the radiological diagnosis of RE. PMID- 16710653 TI - Supportive care needs of people with brain tumours and their carers. AB - GOALS OF WORK: The diagnosis and treatment of a brain tumour may result in long term changes in a patient's functional and social abilities and/or in a greatly reduced life span. A qualitative investigation was conducted to examine the supportive care needs of patients with brain tumour and their carers. MATERIALS AND METHODS: Overall, 18 patients and 18 carers participated in focus groups or telephone interviews, following a structured interview guide to elicit supportive care services of importance to these patients and carers. MAIN RESULTS: Six major themes were identified using the framework analysis method, including needs for information and coping with uncertainty, practical support, support to return to pretreatment responsibilities or prepare for long-term care, support to deal with social isolation and organize respite care, support to overcome stigma/discrimination and support to discuss potentially reduced life expectancy. CONCLUSIONS: Five recommendations to improve service delivery include: assignment of a dedicated member of the care team or case manager; proactive dissemination of information, education and psychosocial support; access to objective assessment of neuropsychological functioning; facilitating easier access to welfare payments; and services facilitating communication about difficult illness related topics. Provision of services along these recommendations could improve supportive care of brain tumour patients and their carers. PMID- 16710659 TI - Dispersal strategies in sponge larvae: integrating the life history of larvae and the hydrologic component. AB - While known to be uniformly non-feeding, short-lived, and potentially short dispersing, sponge larvae display different behaviours (swimming ability and taxis). Our aim was to show whether sponge larvae with different behaviours exhibit different dispersal strategies under variable intensity of water movements. We first assessed the distribution of larvae of six taxa: Dictyoceratida spp., Dysidea avara, Crambe crambe, Phorbas tenacior, Scopalina lophyropoda, and Cliona viridis, collected through plankton sampling, and the abundance of the corresponding adult sponges across three hard bottom communities and a sandy bottom from a north-west Mediterranean rocky shore. We then tested adult-larvae couplings (abundance of larvae vs abundance of adults) under increasing levels of water movements (surge) to assess the importance of this environmental factor in driving differences in dispersal strategies. Adults of Dictyoceratida spp., D. avara, and P. tenacior were most abundant in semi-dark caves (SDC), C. crambe and C. viridis in communities of sciaphilic algae (SA), whereas the distribution of S. lophyropoda was extremely patchy, being present almost only in the SA community of one of the five stations studied. Larvae of Dictyoceratida spp. and P. tenacior were more abundant in the SDC, whereas D. avara and C. crambe were homogeneously distributed across the communities. The larvae of C. viridis were more abundant in the SA communities and the S. lophyropoda larvae were mostly present in one station and one community (SA). Increased water movement did not modify the adult-larvae coupling for Dictyoceratida spp., D. avara, and C. crambe, whereas it broke up the positive association for P. tenacior and to some extent S. lophyropoda. For C. viridis, possible variability in adult-larvae coupling was not tested because the larvae were collected on only one day under calm sea conditions. We confirm that efficient-swimming larvae with some cue response can actively counteract hydrodynamic forces and highlight the importance of both larval behaviour and environmental conditions in determining small-scale patterns of dispersal. PMID- 16710660 TI - Changes in resource concentration and defence during leaf development in a tough leaved (Nothofagus moorei) and soft-leaved (Toona ciliata) species. AB - Developing leaves that are soft, with high concentrations of resources, can be particularly vulnerable to herbivore damage. Since a developing leaf cannot be very tough, given the constraints of cell expansion, the major form of protection is likely to be chemical defence. We investigated changes in concentration of herbivore resources (protein, carbohydrates and water) and putative defences (total phenolics, tannin activity, cyanogenic glycosides, alkaloids, cell wall, and leaf mechanics) across five leaf development stages of the soft-leaved Toona ciliata M. Roem. and the tough-leaved Nothofagus moorei (F. Muell.) Krasser. Chemical defences were predicted to be more highly developed in young than expanded leaves of both species, and to decline more in expanded leaves of N. moorei, which become tough and strong at maturity, than in the softer expanded leaves of T. ciliata. Resources and defences were dynamic within the developing leaves. Highest concentrations of protein were recorded in young leaves in both species, and highest levels of non-structural carbohydrate were recorded in young leaves of T. ciliata. Allocation to defence varied in both amount and type across leaf stages. In T. ciliata, there was an increase in chemical defence in expanded leaves (tannin activity, alkaloids). However, in N. moorei, increasing strength and toughness of developing leaves coincided with decreasing chemical defence, consistent with our hypothesis. For phenolics, this decrease was partly due to dilution by cell wall, but cyanogenic glycosides were present in young leaves and absent in fully mature leaves. These results are consistent with leaf toughness acting as an effective anti-herbivore defence, thereby reducing the need for investment in chemical defence. PMID- 16710663 TI - Localized nasopharyngeal amyloidosis with remarkable early enhancement on dynamic contrast-enhanced MR imaging. PMID- 16710664 TI - Time-resolved contrast-enhanced magnetic resonance angiography of the hand with parallel imaging and view sharing: initial experience. AB - We sought to compare a three-dimensional, contrast-enhanced, magnetic resonance angiogram (3D CE MRA) sequence combining parallel-imaging (generalised autocalibrating partially parallel acquisitions (GRAPPA)) with a time-resolved echo-shared angiographic technique (TREAT) in an intraindividual comparison to a standard 3D MRA sequence. Four healthy volunteers (27-32 years), and 11 patients (11-82 years) with vascular pathologies of the hand were examined on a 1.5-Tesla (T) MR system (Magnetom Avanto, Siemens, Erlangen, Germany) using two multichannel receiver coils. Following automatic injection (flow rate 2.5 cc/s) of 0.1 mmol/kg gadoterate (Dotarem, Guerbet, Roissy, France), 32 consecutive 3D data sets were collected with the TREAT sequence (TR/TE: 4.02/1.31 ms, FA: 10 degrees, GRAPPA acceleration factor: R=2, TREAT factor: 5, voxel size: 1.0 x 0.7 x 1.3 mm(3)) and a T1-wwighted 3D gradient-echo sequence (TR/TE: 5.3/1.57 ms, FA: 30 degrees, GRAPPA acceleration factor: 2, voxel size: 0.71 x 0.71 x 0.71 mm(3,)). MR data sets were evaluated and compared for image quality and visualisation of vascular details. In the volunteer group, all MR imaging was successful while technical problems prevented acquisition of the standard protocol in two patients. For the corresponding segments, the number of visible segments was equal on both sequences. Overall image quality was significantly better on the standard protocol than on the TREAT protocol. TREAT MRA provided functional information in lesions with rapid blood flow, e.g. detection of feeding and draining vessels in an haemangioma. TREAT-MRA is a robust technique that combines morphological and functional information of the hand vasculature and deals with the very special physiological demands of vascular lesions, such as quick arteriovenous transit time. PMID- 16710665 TI - Preoperative volume calculation of the hepatic venous draining areas with multi detector row CT in adult living donor liver transplantation: Impact on surgical procedure. AB - OBJECTIVE: The purpose was to assess the volumes of the different hepatic territories and especially the drainage of the right paramedian sector in adult living donor liver transplantation (ALDLT). METHODS: CT was performed in 40 potential donors of whom 28 underwent partial living donation. Data sets of all potential donors were postprocessed using dedicated software for segmentation, volumetric analysis and visualization of liver territories. During an initial period, volumes and shapes of liver parts were calculated based on the individual portal venous perfusion areas. After partial hepatic congestion occurring in three grafts, drainage territories with special regard to MHV tributaries from the right paramedian sector, and the IRHV were calculated additionally. Results were visualized three-dimensionally and compared to the intraoperative findings. RESULTS: Calculated graft volumes based on hepatic venous drainage and graft weights correlated significantly (r = 0.86, P < 0.001). Mean virtual graft volume was 930 ml and drained as follows: RHV: 680 ml, IRHV: 170 ml (n = 11); segment 5 MHV tributaries: 100 ml (n = 16); segment 8 MHV tributaries: 110 ml (n = 20). When present, the mean aberrant venous drainage fraction of the right liver lobe was 28%. CONCLUSION: The evaluated protocol allowed a reliable calculation of the hepatic venous draining areas and led to a change in the hepatic venous reconstruction strategy at our institution. PMID- 16710667 TI - Postoperative retinal break after 25-gauge transconjunctival sutureless vitrectomy: report of four cases. AB - BACKGROUND: A 25-gauge transconjunctival sutureless vitrectomy (TSV) has been reported effective. However, complications such as postoperative retinal detachment have been reported. In this study, we report four cases of retinal breaks found after 25-gauge TSV. In this study, we investigated factors contributing to occurrence of postoperative complications. METHODS: Seventy-five patients (75 eyes) underwent 25-gauge TSV surgery at Kanazawa University hospital between April 2004 and September 2005. Postoperative follow-up monitoring was done for at least 3 months. The surgical charts were reviewed. RESULTS: Retinal breaks not accompanied by retinal detachment were noted postoperatively in four patients. All four of these patients had preoperative idiopathic macular holes. In all cases, there was no vitreous traction around the retinal break and photo coagulation was performed. One eye with age-related macular degeneration developed intraoperative rhegmatogenous retinal detachment. No other complications were observed during the intraoperative and postoperative periods. CONCLUSION: Upon performing 25-gauge TSV for macular hole repair, care should be taken to detect retinal breaks and retinal detachment intraoperatively and postoperatively. PMID- 16710666 TI - Is microbubble-enhanced ultrasonography sufficient for assessment of response to percutaneous treatment in patients with early hepatocellular carcinoma? AB - The objective of this study was to assess the efficacy of contrast-enhanced ultrasonography (CEUS) with SonoVue to evaluate the response to percutaneous treatment (ethanol injection/radiofrequency) of hepatocellular carcinoma in comparison with spiral computed tomography (CT) immediately and 1 month after treatment. Forty-one consecutive cirrhotic patients with early stage tumor (not suitable for resection) were included. Spiral CT and CEUS were performed in all patients before treatment, in the following 24 h, and 1 month later. The results of each examination were compared with the 1-month spiral CT, considered the gold standard technique. The 24-h CEUS and the 24-h spiral CT sensitivity to detect residual disease were 27% and 20%, respectively. The 24-h CEUS and the 24-h spiral CT positive predictive value of persistent vascularization detection were 75% and 66%, respectively. The 1-month CEUS detected partial responses in ten out of 11 cases (91% sensitivity, 97% specificity, 95% accuracy). Spiral CT and CEUS performed in the 24 h following treatment are slightly useful to evaluate therapeutic efficacy. The 1-month CEUS has a high diagnostic accuracy compared with spiral-CT in the usual assessment of percutaneous treatment response. PMID- 16710668 TI - Other therapies for BPH patients: desmopressin, anti-cholinergic, anti inflammatory drugs, and botulinum toxin. AB - The usual treatments of benign prostate hyperplasia (BPH) including the alpha blockers, the inhibitors of the 5-alpha reductase and the phytotherapy drugs allow significant improvements of the lower urinary tracts symptoms (LUTS). However, some patients are not responders or have side effects due to the treatments. Other therapeutic approaches described in the literature are possible in order to alleviate the LUTS. The anti-cholinergic drugs seem to be efficient against the irritating symptoms even if they are supposed to be contra-indicated when there is BPH. Anti-diuretic hormone could be useful to treat nocturia due to diuresis reversal. Inflammation is a part of the underlying mechanisms of BPH and as such the role of the anti-inflammatory drugs has to be revised. Eventually, botulinum toxin is more and more used for patients with neurological bladder and could also have a role in LUTS. If the coming clinical studies on those different treatments confirm the preliminary results, the learning societies in charge of the guidelines would have to update the decision trees by adding these new therapeutic approaches. PMID- 16710669 TI - First-line treatment for symptomatic benign prostatic hyperplasia: is there a particular patient profile for a particular treatment? AB - When the range of treatment options for benign prostatic hyperplasia (BPH) is as broad as the BPH spectrum of symptoms, how should urologists and their patients choose the best initial treatment? Treatment goals should include reducing both lower urinary tract symptoms and relieving associated morbidities, such as urinary retention, persistent gross hematuria, recurrent infections, bladder stones, or renal insufficiency-which are all indications for surgery. However, if one views BPH as a chronic and progressive disease, should a primary goal of BPH management be the prevention of the BPH-associated morbidities? The goal of this paper is to provide a literature update regarding various treatment options in the setting of initial treatment for symptomatic BPH. Novel approaches to BPH management are discussed. In all cases, the risks and benefits of each treatment need to be considered and discussed with the patient; the decision is ultimately up to the patient and his urologist. PMID- 16710671 TI - Nasal NO measurement by direct sampling from the nose during breathhold: Aspiration flow, nasal resistance and reproducibility. AB - The objective of this study was assessment of the effect of aspiration flow, the nasal cycle, and time on nasal nitric oxide (nNO) concentrations in air sampled from one nostril during breathhold. nNO was measured in 45 healthy subjects (19 males, aged 18-45 years) from one nostril during breathholding. We compared nNO values and time to plateau in both nostrils with 3 aspiration flows (280, 700, 1,200 ml/min) and assessed the short-term and long-term reproducibility. Mean nNO values at flows of 280, 700 and 1,200 ml/min differed significantly (P < 0.01): 854, 474, 380 ppb, respectively. The (median) plateau was reached after 6, 4 and 3 s for the different flows. The within-subject coefficient of variability was always < 5%. We found no difference in nNO between left-, right-, largest or smallest nostril (P > 0.10). nNO values after 6, 24 h and 7 days were not significantly different from baseline (P > 0.10) and showed fair reproducibility. The highest aspiration flow was experienced as unpleasant. nNO can be measured in either nostril and shows no diurnal variation. The measurement is quick, reproducible, feasible and best accepted with an aspiration flow of 700 ml/min during breathhold for 10 s. PMID- 16710672 TI - Association of cigarette smoking and tar and nicotine intake with development of type 2 diabetes mellitus in men and women from the general population: the MONICA/KORA Augsburg Cohort Study. AB - AIMS/HYPOTHESIS: We examined sex-specific associations between cigarette smoking and incident type 2 diabetes mellitus in Germany. SUBJECTS, MATERIALS AND METHODS: The study was based on 5,470 men and 5,422 women (aged 25-74 years) without diabetes who participated in one of the three population-based MONICA Augsburg surveys between 1984 and 1995. Incident cases of type 2 diabetes were assessed using follow-up questionnaires. Hazard ratios (HRs) were estimated from Cox proportional hazard models. RESULTS: Up to 31 December 2002 a total of 409 cases of incident type 2 diabetes among men and 263 among women were registered. The number of cigarettes and the nicotine and tar consumption per day were associated with a significantly increased risk of type 2 diabetes among men, but not among women; this could be due to the low power of the study in women. After multivariable adjustment, the HRs for type 2 diabetes compared with never-smokers were 1.48, 2.03 and 2.10 for men smoking 1 to 14, 15 to 19 and > or =20 cigarettes/day (p for trend <0.0001) and 1.25, 1.34 and 1.37 for women smoking 1 to 9, 10 to 19 and > or =20 cigarettes/day (p for trend 0.0985). Compared with never-smokers, the HRs for increasing tar intake in men (1-167, 168-259 and > or =260 mg/day) were 1.45, 2.32 and 2.07 (p for trend <0.0001); the respective HRs in women (1-89, 90-194 and > or =195 mg/day) were 1.18, 1.57 and 1.24 (p for trend 0.1159). CONCLUSIONS/INTERPRETATION: Cigarette smoking is an important modifiable risk factor of type 2 diabetes particularly in men from the general population. PMID- 16710673 TI - Long-term, low-maintenance storage of Acanthamoeba strains. AB - A simple, low cost method is described for the long-term storage of the potential human pathogen Acanthamoeba. Amoebae are placed in dried, sterile clay pellets within which they form cysts that remain viable and pure for at least 5 years. This method compares favorably with other storage methods and requires no specialist equipment. We have demonstrated that amoeba stored by this method retain both their pathogenic potential and their small subunit ribosomal DNA sequence type. PMID- 16710676 TI - [Future of chemotherapy]. AB - The future development of chemotherapy is derived based upon recent advances. With regard to adjuvant therapy a trend towards standardized prediction of recurrence risk using all available prognostic markers (e.g., nomograms) is observed. Furthermore, in some tumors major progress has been made regarding the development of "molecular classifiers" defining tumor biology (predicting clinical outcome) by analysis of molecular changes. In adjuvant therapy considerable advances may be achieved by use of new chemotherapeutic agents as well as sequential, dose-dense and dose-intensified regimens. However, in metastatic disease no breakthrough can be expected at least with regard to survival suggesting that quality of life needs to be addressed with more emphasis. Using targeted drugs alone or in combination with chemotherapy advances concerning adjuvant therapy as well as metastatic disease are observed. Further targeted drugs have entered clinical development. However, clarification of the relation between detection of the target(s) and drug activity will fundamentally change current treatment concepts. PMID- 16710674 TI - Analysis of P-glycoprotein expression in purified parasite plasma membrane and food vacuole from Plasmodium falciparum. AB - A P-glycoprotein homologue (Pgh1) is believed to play a role in modulating levels of chloroquine resistance in Plasmodium falciparum. To study the role of Pgh1 in the mechanism of chloroquine (CQ) resistance, antisera were raised against this protein. There was no direct association between the level of Pgh1 expression and chloroquine sensitivity. We also failed to detect phosphorylation of Pgh1 in the food vacuole (FV), suggesting that other mechanisms regulate the chloroquine resistant (CQR) phenotype. Therefore, high levels of expression of Pgh1 or phosphorylation of this protein in the FV could not account for CQ sensitivity. In addition, the lack of inhibition of CQ accumulation by anti-Pgh1 antibodies suggests that Pgh1 is not involved as a CQ transporter in the plasma membrane of P. falciparum. Furthermore, resistance reversers do not appear to act at the plasma membrane level. PMID- 16710677 TI - [Therapeutic options for hormone-refractory prostate cancer]. AB - For a long time, hormone-refractory prostate cancer was regarded as a chemoresistant tumor. The introduction of taxanes has prompted a change in this opinion. For the first time treatment with 75 mg/m(2) docetaxel every 3 weeks has evidenced a survival benefit in a phase III trial (median survival of 18.9 months versus 16.5 months with mitoxantrone). Further advantages were improved pain reduction and quality of life. Neutropenia was foremost among the side effects. Docetaxel is currently the standard treatment for hormone-refractory prostate cancer. The morbidity of metastatic hormone-refractory prostate cancer is influenced by bone metastases. Pain is a prominent feature. Skeletal complications are frequent. Therapy with 4 mg zoledronic acid reduced skeletal complications significantly in comparison to placebo. The most pronounced effect is the reduction of pathological fractures. Side effects include flu-like symptoms, muscle pain, and edemas. Zoledronic acid also belongs to the standard treatment of hormone-refractory prostate cancer with bone metastases. PMID- 16710678 TI - [Value of systemic chemotherapy in bladder cancer]. AB - Almost half of the patients with muscle invasive disease already harbor at the time of their first diagnosis occult or distant metastases. Systemic disease has a poor prognosis with a long term survival of less than 10%. The administration of systemic chemotherapy aims to improve the course of locally advanced or metastatic disease.A survival benefit of 5% for patients receiving neoadjuvant and 9-11% using adjuvant chemotherapy is in the first scenario minimal, in the adjuvant setting to be noteworthy. The MVAC-schedule and the Gemcitabine/Cisplatin-combination chemotherapy have to be regarded as standard for induction chemotherapy. However, the 5-year survival rates with 15 or 13% are disappointing.Thus, prognostic factors gain importance since with their consideration significant differences in survival rates can be found. Hope is provided by a novel class of substances, the target-specific drugs, which selectively interfere with the cascade of steps involved in tumorigenesis. PMID- 16710679 TI - [Metastases with CUP syndrome]. AB - Carcinoma of unknown primary is common, accounting for 2-6% of all cancer patients. The primary site is found in less than 25% of patients before death and frequently goes undiscovered at postmortem examination. At the time point of first diagnosis of CUP syndrome, usually more than 80% of the patients present a disseminated situation. Prognosis depends on the involved site and is unaffected by whether or not the primary site is ever found. For patients presenting with metastasis to peripheral lymph nodes, node dissection may be curative. In patients with small cell malignancies, peritoneal carcinomatosis (in women), poorly differentiated carcinomas involving external lymph nodes, mediastinum, or retroperitoneum, but without metastases to viscera or bone, objective long-term responses are possible with combination chemotherapy. For all other patients, toxic therapies are recommended only for patients with good functional status, for palliation of symptoms when they develop, and for continuous support of the quality of life. PMID- 16710680 TI - [Effect mechanism of intravesical BCG immunotherapy of superficial bladder cancer]. AB - Immunotherapy for treatment of solid cancer mostly is an experimental treatment. In contrast, intravesical immunotherapy of superficial bladder cancer with bacille Calmette-Guerin (BCG) is clinically well established and accepted worldwide because of better results compared to topical chemotherapy. BCG is currently regarded as the most successful immunotherapy of cancer. Unfortunately the mechanism of action has not yet been fully clarified. This article gives an overview on the complex research on the mechanisms of actionhighly successful therapy. PMID- 16710681 TI - Muscle synergies during voluntary body sway: combining across-trials and within-a trial analyses. AB - We investigated co-varied changes in muscle activity during voluntary sway tasks that required a quick shift of the center of pressure (COP). We hypothesized that multi-muscle synergies (defined as task-specific covariation of elemental variables, muscle modes) stabilize a COP location in the anterior-posterior direction prior to a voluntary COP shift and that during the shift the synergies would weaken. Standing subjects performed two tasks, a cyclic COP shift over a range corresponding to 80% of the maximal amplitude of voluntary COP shift at 1 Hz and a unidirectional quick COP shift over the same nominal amplitude. The cyclic sway task was used to define muscle modes (M-modes, leg and trunk muscle groups with parallel scaling of muscle activation level within a group) and the relations between small changes in the magnitudes of M-modes [in the principal component analysis (PCA), the M-mode magnitudes are equivalent to PC scores] and COP shifts. A novel approach was used involving PCA applied to indices of muscle integrated activity measured both within a trial and across trials. The unidirectional sway task was performed in a self-paced (SP) manner and under a typical simple reaction time (RT) instruction. M-modes were also defined along trials at those tasks; they have been shown to be similar across tasks. Integrated indices of muscle activity in the SP-sway and RT-sway tasks were transformed into the M-modes. Variance in the M-mode space was partitioned into two components, one that did not affect the average value of COP shift (V (UCM)) and the other that did (V (ORT)). An index (DeltaV) corresponding to the normalized difference between V (UCM) and V (ORT) was computed. During steady state posture, DeltaV was positive corresponding to most M-mode variance lying in a sub-space corresponding to a stable COP location across trials. Positive DeltaV values have been interpreted as reflecting a multi-M-mode synergy stabilizing the COP location. The magnitude of DeltaV was larger in SP trials than in RT trials. During voluntary COP shifts, the DeltaV magnitude dropped to zero or even became negative. We conclude that M-mode synergies stabilize COP location during quiet standing, while these synergies weaken or disappear during fast voluntary COP shifts. Under RT conditions, the COP stabilizing synergies were weaker supposedly to facilitate a quick COP shift without time for preparation. The suggested method of M-mode identification may potentially be applied to analysis of postural synergies in persons with impaired postural control such as elderly persons, persons with atypical development, or in the course of rehabilitation after an injury. PMID- 16710682 TI - Increased cannabinoid receptor density in the posterior cingulate cortex in schizophrenia. AB - The posterior cingulate cortex (PCC) has recently been implicated in the pathophysiology of schizophrenia, through both animal and human studies. We have recently shown abnormal glutamate, GABA, and muscarinic receptor binding in the PCC in schizophrenia. In addition, there is evidence for an abnormal endogenous cannabinoid system in schizophrenia. The endogenous cannabinoid system, including CB1 receptors, is proposed to play a role in modulating neurotransmission via affecting the release of a variety of neurotransmitters, (e.g. GABA). In the present study, we used quantitative autoradiography to investigate the binding of [(3)H]CP-55940 to CB1 receptors in the PCC in schizophrenia subjects compared to controls. A significant 25% increase in CB1 binding was found in the superficial layers (layer I, II) of the PCC of schizophrenia subjects compared to controls, none of whom had recently used cannabis. There was no statistical difference in CB1 binding in the deeper layers (layers III-VI) between the two groups. There were no significant correlations between CB1 binding density and age, PMI, pH, brain weight, freezer storage time, or final recorded antipsychotic drug dose. These results show an increase in CB1 receptor density in the PCC in schizophrenia, and therefore provide support for a role of the endogenous cannabinoid system in schizophrenia. PMID- 16710683 TI - The impact of restricted diet on brain function using BOLD-fMRI. AB - We investigated the effect of a restricted diet model on activity in the human motor cortex using functional magnetic resonance imaging (fMRI). Two series of blood oxygenation level-dependent (BOLD)-fMRI measurements were made in healthy subjects performing simple motor tasks using their right hands. The first series was done 5-10 days prior to the restricted diet schedule (controls), and the second series was performed after 25-28 days of restricted diet, in the form of a religious fast (Ramadan). The size and intensity of the activated area in the motor cortex increased during the time of restricted diet versus the controls. We conclude that restricted diet has a significant effect on cerebral activity, as shown by BOLD-fMRI, although the exact relationship between the images and neuronal activity due to the restricted diet is still to be determined. PMID- 16710685 TI - Grasp effects of the Ebbinghaus illusion are ambiguous. AB - The assumption that the Ebbinghaus/Titchener illusion deceives perception but not grasping, which would confirm the two-visual-systems hypothesis (TVSH) as proposed by Milner and Goodale (The visual brain in action, 1995), has recently been challenged. Franz et al. (Exp Brain Res 149:470-477, 2003) found that the illusion affects both perception and grasping, and showed that the effect of the illusion on the peak grip aperture (PGA) cannot be accounted for by different sizes of the gap that separates the central target disk from the surrounding flankers. However, it is not yet clear if the presence of flankers per se influences grasping. We therefore compared kinematic parameters of prehension, using the Ebbinghaus illusion, and a neutral control condition where normal subjects grasped a disk without any flankers. In accordance with the well-known effects of the illusion on perceived size, the PGA was smaller when the target disk was surrounded by large flankers, and larger when it was encircled by small flankers. However, the largest PGA values were reached in the neutral control condition. Hence the presence of flankers leads to a general reduction of the PGA, possibly because the flankers are regarded as obstacles. This 'reduction effect' casts doubts on how appropriate it is to directly compare perceptual measures and PGA values when using the Ebbinghaus illusion. Even smaller effects of the illusion on the PGA compared to larger perceptual effects cannot be unequivocally interpreted. PMID- 16710684 TI - The role of prefrontal cortex in visuo-spatial planning: A repetitive TMS study. AB - The visuo-spatial planning process is based on an "opportunistic" combination of heuristics and strategies, carried out in small units during the execution of plans. In order to investigate the functional role of the prefrontal cortex in heuristic switching, 42 healthy controls performed a labyrinth crossing task (the Maps Test). During this computerized version of the Travelling Salesperson Problem, subjects had to decide which order of locations optimizes total travel time and distance. This task was performed with and without 1 Hz repetitive transcranial magnetic stimulation (rTMS), which exerts an inhibitory action on the targeted area, applied during the task over bilateral frontal sites (active stimulation) and parieto-occipital site (sham stimulation). Only repetitive bilateral rTMS over F3 and F4 significantly decreased the number of strategies with changes of heuristics, and increased the number of movements required to solve the task. This behaviour contrasts with the performance of healthy subjects in the planning task, but is consistent with the performance of frontal traumatic brain injury patients. The results indicate that, in a visuo-spatial problem solving task, the prefrontal cortex is involved in the switching between heuristics during the execution of a plan. PMID- 16710693 TI - Environmental analysis of fluorinated alkyl substances by liquid chromatography (tandem) mass spectrometry: a review. AB - Fluorinated alkyl substances (FASs) are widely distributed contaminants that have been found in many environmental, human and biological samples throughout the world. Perfluorochemicals are used in many industry and consumer products, such as polymers and surfactants, because they have unique and useful properties (they are stable, chemically inert and generally unreactive). However, these compounds have also been found to be toxic, persistent and bioaccumulative. In recent years various analytical methods have been developed for the analysis of FASs in environmental samples. Most of these methods are based on liquid chromatography coupled to mass spectrometry (LC-MS) or tandem mass spectrometry (LC-MS/MS), since this is considered to be the technique of choice. This article reviews the various LC-(tandem)MS methods described so far for the analysis of FASs in water, sediment, sludge and biota samples. It discusses the main experimental conditions used for sample pretreatment and for analysis as well as the most relevant problems encountered and the limits of detection achieved. PMID- 16710695 TI - Optimization of an analytical methodology for the determination of alkyl- and methoxy-phenolic compounds by HS-SPME in biomass smoke. AB - A sampling and analysis method for the determination of 21 phenolic compounds in smoke samples from biomass combustion has been developed. The smoke is used to make smoked foods, following an artisanal procedure used in some parts of the Canary Islands. The sampling system consists of a Bravo H air sampler, two impingers, each one containing an aqueous solution of sodium hydroxide 0.1 mol L( 1), followed by a silica gel trap. The variables optimized to reach the best sampling conditions were volume of absorbent solution and sampling flow. Under the optimum conditions, 100 mL of absorbent solution of NaOH 0.10 mol L(-1) and 2 L min(-1) for the sampling flow, sampling efficiencies are higher than 80%. Analysis of phenolic compounds was carried out by headspace solid-phase microextraction (HS-SPME) coupled to gas chromatography-mass spectrometry (GC MS). Five different fiber coatings were employed in this study. By means of a central composite design, extraction time, salt concentration, and pH of the solution were optimized: 65-microm carbowax-divinylbenzene, extraction time 90 min, concentration in NaCl of 35% (m/v), and pH 2 yielded the highest response. Detection limits of phenol and their alkyl derivatives, guaiacol and eugenol, are between 1.13 and 4.60 ng mL(-1). 3-Methoxyphenol, 2,6-dimethoxyphenol, and vanillin have detection limits considerably higher. Good linearity (R2 > or = 0.98) was observed for all calibration curves in the established ranges. The reproducibility of the method (RSD, relative standard deviation) was found to oscillate between 7 and 18% (generally close or lower than 10%). PMID- 16710696 TI - Repeated 28-day oral toxicity study of ketoconazole in rats based on the draft protocol for the "Enhanced OECD Test Guideline No. 407" to detect endocrine effects. AB - We performed a 28-day repeated-dose toxicity study of ketoconazole, a widely used an antimycotic drug, based on the draft protocol of the "Enhanced OECD Test Guideline 407" (Enhanced TG407) to investigate whether ketoconazole has endocrine mediated properties according to this assay. Seven-week-old SD rats were administered with ketoconazole daily by oral gavage at doses of 0, 6.25, 25 or 100 mg kg(-1) day(-1) for at least 28 days. The ketoconazole-treated male rats showed reduction of epididymis and accessory sex organ weights, spermatid retention in the seminiferous tubules, decrease of testosterone and increases of estradiol, luteinizing hormone (LH) and follicular stimulating hormone (FSH). A prolongation of the estrous cycle and increases of estradiol, LH and FSH were observed in the treated female rats. Thyroxin and triiodothyronine were decreased and thyroid-stimulating hormone was increased in both sexes; however, there were no compound-related microscopic lesions in the thyroid gland or changes in the thyroid weight. The endocrine-related effects of ketoconazole could be detected by the parameters examined in the present study based on the Organization for Economic Cooperation and Development (OECD) protocol, suggesting that the Enhanced TG407 protocol should be a suitable screening test for detection of endocrine-mediated effects of chemicals. PMID- 16710698 TI - In ovo exposure quail assay for risk assessment of endocrine disrupting chemicals. AB - Although there are in vivo assays using various organisms for the risk assessment of chemicals with endocrine disrupting properties, effective experimental methods for avian species are still under debate. We have developed an in ovo exposure assay using Japanese quail eggs, aimed at assessing disrupting effects on avian reproductive development and function. Hybrid eggs from Brazilian Brown male and White Egg female quails, which can be genetically sexed by their plumage color after hatching, were prepared, and test materials dissolved in olive oil were injected into the air-chamber on day 10 of incubation. After sexual maturation of hatched chicks, we observed egg production by females and the egg quality and male-typical reproductive behavior, and then examined reproductive system morphology and serum steroid concentrations in both sexes. Treatment with a synthetic estrogen, diethylstilbestrol (DES, 0.5-50 ng/g egg), dose-dependently reduced the eggshell thickness and strength of eggs. A few females treated with 5 ng/g DES per egg produced soft-shelled/ unmarked eggs, and all laying females treated with 50 ng/g egg produced eggs completely lacking shells. DES also induced shortening of the left oviduct and abnormal development of the right oviduct in a dose-dependent manner, while testis weight was reduced symmetrically. In addition, 2,2',4',6'-tetrachlorobiphenyl-4-ol (10-1,000 ng/g egg), which previously showed relatively high estrogenic activity in vitro, caused dose-dependent shortening of the left oviduct and reduction in testis weight. The methods for evaluating endocrine disrupting effects and preparing experimental birds proposed in the present study are expected to facilitate assays for avian reproductive toxicology. PMID- 16710697 TI - Irritative effects of fumes and aerosols of bitumen on the airways: results of a cross-shift study. AB - Possible health hazards of fumes and aerosols of bitumen are in discussion, and data on their adverse effects on human airways under current exposure conditions are limited. To assess the irritative effects of exposure to fumes and aerosols of bitumen on the airways, a cross-sectional cross-shift study was conducted including external and internal exposure measurements, spirometry and especially non-invasive methods like nasal lavage collection and induction of sputum in order to identify and evaluate more precisely inflammatory process in the upper and lower airways. The cross-shift study comprised 74 mastic asphalt workers who were exposed to fumes and aerosols of bitumen and 49 construction workers without this exposure as reference group. Questionnaire, spirometry, ambient monitoring and urinary analysis were performed. Humoral and cellular parameters were measured in nasal lavage fluid (NALF) and induced sputum. For data analysis, a mixed linear model was performed on the different outcomes with exposure group, time of measurement (pre-, post-shift), current smoking, German nationality and age as fixed factors and subjects as random factor. Based on personal exposure measurements during shift, mastic asphalt workers were classified into a low (< or =10 mg/m(3); n = 46) and a high (>10 mg/m(3); n = 28) exposure group. High exposure was accompanied by significant higher urinary post-shift concentrations of 1-hydroxypyrene and the sum of hydroxyphenanthrenes. Acute respiratory symptoms were reported more frequently in the high exposure group after shift. Significant cross-shift declines in lung function parameters (forced expiratory volume in 1 s [FEV(1) (% predicted)] and forced vital capacity [FVC (% predicted)]) were measured in mastic asphalt workers. Pre-shift FEV(1) (% predicted) and FVC (% predicted) were higher in the low exposure group. In pre- and post-shift NALF samples, interleukin (IL)-1beta-, IL-8- and total protein concentrations were lower in the low exposure group compared to the reference and the high exposure group. Pre- and post-shift neutrophil percentages in both nasal and sputum samples were also lower in the low exposure group. Significantly higher pre- and post-shift sputum concentrations of IL-8, IL-6, nitrogen oxide (NO) derivatives and total protein were detected especially in highly exposed workers. Irritative effects of exposure to fumes and aerosols of bitumen on the upper and lower airways were apparent, especially in mastic asphalt workers with exposure above 10 mg/m(3). PMID- 16710699 TI - Temperature-dependent oxygen extraction from the ventilatory current and the costs of ventilation in the cephalopod Sepia officinalis. AB - Earlier work found cuttlefish (Sepia officinalis) ventilatory muscle tissue to progressively switch to an anaerobic mode of energy production at critical temperatures (T (c)) of 7.0 and 26.8 degrees C. These findings suggested that oxygen availability limits thermal tolerance. The present study was designed to elucidate whether it is the ventilatory apparatus that sets critical temperature thresholds during acute thermal stress. Routine metabolic rate (rmr) rose exponentially between 11 and 23 degrees C, while below (8 degrees C) and above (26 degrees C) this temperature range, rmr was significantly depressed. Ventilation frequency (f (V)) and mean mantle cavity pressure (MMP) followed an exponential relationship within the entire investigated temperature range (8-26 degrees C). Oxygen extraction from the ventilatory current (EO(2)) decreased in a sigmoidal fashion with temperature, falling from > 90% at 8 degrees C to 32% at 26 degrees C. Consequently, ventilatory minute volume (MV(V)) increased by a factor of 20 from 7 to 150% body weight min(-1) in the same temperature interval. Increases in MMP and MV(V) resulted in ventilatory muscle power output (P (out)) increasing by a factor of > 80 from 0.03 to 2.4 mW kg(-1) animal. Nonetheless, costs for ventilatory mechanics remain below 1.5% rmr in the natural thermal window of the population (English Channel, 9-17 degrees C), owing to very low MMPs of < 0.05 kPa driving the ventilatory stream, and may maximally rise to 8.6% rmr at 26 degrees C. Model calculations suggest that the ventilatory system can maintain high arterial PO(2) values of > 14 kPa over the entire temperature interval. We therefore conclude that the cuttlefish ventilation system is probably not limiting oxygen transfer during acute thermal stress. Depression of rmr, well before critical temperatures are being reached, is likely caused by circulatory capacity limitations and not by fatigue of ventilatory muscle fibres. PMID- 16710700 TI - Unchanged expression of the sodium-dependent phosphate cotransporter NaPi-IIa despite diurnal changes in renal phosphate excretion. AB - Renal phosphate excretion is subjected to circadian rhythmicity. The bulk of filtered inorganic phosphate (P(i)) is reabsorbed by the sodium-dependent phosphate cotransporter NaPi-IIa. The regulation of proximal tubular phosphate reabsorptive capacity is largely attributed to the altered abundance of NaPi-IIa residing in the brush border membrane (BBM) of proximal tubular cells. Therefore, we examined if the diurnal rise in renal phosphate excretion is accompanied by a corresponding change in NaPi-IIa expression. Renal phosphate excretion, creatinine clearance, and serum phosphate were determined at consecutive time points in rats, starting from 8 a.m. until 5 p.m. During this period, renal phosphate excretion (fractional P(i) excretion) increased more than eightfold until 5 p.m. compared to the morning values at 8 a.m. In addition, serum phosphate and creatinine clearance as well as the calculated tubular phosphate threshold increased. Neither immunoblot analysis of BBMs nor immunohistochemical staining for NaPi-IIa yielded evidence for a lower abundance of NaPi-IIa in kidneys collected in the afternoon compared to those in the morning. However, kidneys sampled in the afternoon showed a small decrease (14%) in (32)P uptakes into BBM vesicles (BBMVs). Thus, the diurnal rise in renal phosphate excretion was associated with a mild reduction in the sodium-dependent phosphate transport rate in proximal tubular BBMs. There was no apparent downregulation of NaPi-IIa abundance and only a small reduction in Na(+)-dependent Pi-transport activity. Thus, the diurnal changes in urinary phosphate excretion appear to be mainly related to changes in serum phosphate and tubular threshold but not to NaPi-IIa expression. PMID- 16710702 TI - No association between complement factor H gene polymorphism and exudative age related macular degeneration in Japanese. AB - Age-related macular degeneration (ARMD) is the leading cause of blindness in the elderly population not only Western but also Asian industrial countries. In Caucasian, a polymorphism of the complement factor H gene (CFH), the C allele of rs1061170 (Y402H), was established as the first strong genetic factor for excursively exudative type of ARMD. In this study, we performed an extensive sequencing of the 22 exons in the CFH gene by recruiting 146 exudative ARMD patients and 105 normal controls of Japanese origin and identified 61 polymorphisms. We found that the frequency of the C allele of rs1061170 (Y402H) is much lower (0.04) in Japanese controls than in Caucasians (0.45). No case disease susceptibility to exudative ARMD was noted for rs1061170 (Y402H) (chi (2) = 3.19, P (corr) = 0.423), or other 12 single nucleotide polymorphisms (SNPs) whose frequency is greater than 0.05. When haplotypes were inferred for 13 SNPs (these 12 SNPs with a frequency greater than 0.05 and rs1061170), three haplotypes whose pattern was similar to those in Caucasians were identified but with substantial difference in frequency. Again we failed to identify genetic association between Japanese exudative ARMD and any of the haplotypes including the J1 haplotype which was shown to be susceptible to ARMD in Caucasians (chi (2 )=( )3.92, P (corr) = 0.157). CFH does not appear to be a primary hereditary contributor to ARMD in Japanese. The absence of CFH contribution to ARMD in Japanese may correlate with the findings in ethnic differences of ARMD phenotypes. PMID- 16710701 TI - Intracellular peptide transporters in human--compartmentalization of the "peptidome". AB - In the human genome, the five adenosine triphosphate (ATP)-binding cassette (ABC) half transporters ABCB2 (TAP1), ABCB3 (TAP2), ABCB9 (TAP-like), and in part, also ABCB8 and ABCB10 are closely related with regard to their structural and functional properties. Although targeted to different cellular compartments such as the endoplasmic reticulum (ER), lysosomes, and mitochondria, they are involved in intracellular peptide trafficking across membranes. The transporter associated with antigen processing (TAP1 and TAP2) constitute a key machinery in the major histocompatibility complex (MHC) class I-mediated cellular immune defense against infected or malignantly transformed cells. TAP translocates the cellular "peptidome" derived primarily from cytosolic proteasomal degradation into the ER lumen for presentation by MHC class I molecules. The homodimeric ABCB9 (TAP-like) complex located in lysosomal compartments shares structural and functional similarities to TAP; however, its biological role seems to be different from the MHC I antigen processing. ABCB8 and ABCB10 are targeted to the inner mitochondrial membrane. MDL1, the yeast homologue of ABCB10, is involved in the export of peptides derived from proteolysis of inner-membrane proteins into the intermembrane space. As such peptides are presented as minor histocompatibility antigens on the surface of mammalian cells, a physiological role of ABCB10 in the antigen processing can be accounted. PMID- 16710708 TI - Comparison of dose conversion factors for radon progeny from the ICRP 66 regional model and an airway tube model of tracheo-bronchial tree. AB - Current epidemiological approaches to radon dosimetry yield a dose conversion factor (DCF) of 4 mSv WLM(-1) while the dosimetric approaches give a value closer to 13 mSv WLM(-1). The present study investigated whether the application of compartment models for the bronchial (BB) and bronchiolar (bb) regions, rather than more anatomically realistic airway tube models, has brought the dosimetric DCF to the higher values. The airway tube model of the tracheo-bronchial tree was used to calculate the effective dose per unit radon exposure. All other elements of the human respiratory tract from the reports of the ICRP or NRC were adopted. A dosimetric derivation of the radon DCF using the airway tube model yielded a value of 14.2 mSv WLM(-1). This value is slightly larger than, but not significantly different from, the result obtained through the ICRP 66 approach. It is concluded that utilization of the airway tube model instead of the regional ICRP 66 compartmental model cannot reconcile the gap between dose conversion factors derived from epidemiological and dosimetric approaches. PMID- 16710703 TI - Improved Agrobacterium-mediated transformation of three maize inbred lines using MS salts. AB - Transformation technology as a research or breeding tool to improve maize is routinely used in most industrial and some specialized public laboratories. However, transformation of many inbred lines remains a challenging task, especially when using Agrobacterium tumefaciens as the delivery method. Here we report success in generating transgenic plants and progeny from three maize inbred lines using an Agrobacterium-mediated standard binary vector system to target maize immature embryos. Eleven maize inbred lines were pre-screened for transformation frequency using N6 salts. A subset of three maize inbred lines was then systematically evaluated for frequency of post-infection embryogenic callus induction and transformation on four media regimes: N6 or MS salts in each of two distinct media backgrounds. Transgenic plants recovered from inbred lines B104, B114, and Ky21 were analyzed for transgene integration, expression, and transmission. Average transformation frequencies of 6.4% (for B104), 2.8% (for B114), and 8% (for Ky21) were achieved using MS salts. Availability of Agrobacterium-mediated maize inbred line transformation will improve future opportunities for maize genetic and functional genomic studies. PMID- 16710709 TI - Do work-related physical factors predict neck and upper limb symptoms in office workers? AB - OBJECTIVES: The objective of this study was to examine the influence of physical exposure at work on neck and upper limb symptoms in office workers. METHODS: Data were used from a prospective cohort study with a follow-up period of 3 years. Independent variables were physical exposure at work, observed and self-reported. Outcome measures were neck-shoulder symptoms and elbow-wrist-hand symptoms. Data were analyzed with the generalized estimating equation (GEE) method, with adjustment for age, gender, psychosocial work characteristics and the outcome at baseline. RESULTS: Neck rotation was associated with neck-shoulder symptoms in the analyses with observed data (OR: 1.57; CI: 0.99-2.50) as well as those with self-reported data (OR: 1.43; CI: 1.02-2.01). Neck extension was also statistically significantly associated with neck-shoulder symptoms (OR: 2.42; CI: 1.22-4.80), but only self-reported data were available. Neck flexion, self reported wrist pronation, self-reported arm elevation and self-reported duration of computer work, were not associated with symptoms. An indication was found of an adverse effect on neck-shoulder symptoms of long working days and on elbow wrist-hand symptoms of self-reported wrist flexion and full-time work or longer compared to part-time work. CONCLUSIONS: Only a limited number of work-related physical factors were related to neck and upper limb symptoms in office workers. Only neck rotation and self-reported neck extension were identified as risk factors for neck-shoulder symptoms. PMID- 16710710 TI - Sick leave as a predictor of job loss in patients with chronic arthritis. AB - OBJECTIVES: To study the occurrence and duration of sick leave as potential risk factors for permanent job loss after 24 months among 112 individuals with chronic arthritis and a disease related problem at work. METHODS: Data collection was embedded in a multicentre randomised controlled trial in which the cost effectiveness of a multidisciplinary job retention vocational rehabilitation programme for employees with chronic arthritis and a disease related problem at work was compared to usual outpatient care. Sick leave (complete or partial) was defined as absenteeism reported to the employer and permanent job loss as receiving a full work disability pension or unemployment. The association between sick leave at baseline and job loss after 24 months was investigated by multivariate logistic regression analysis, including those variables that were univariately significantly associated with job loss after 24 months. RESULTS: At baseline, 60 of the 112 subjects (54%) were on sick leave, with a mean duration of 18.7 weeks, in half of these patients the sick leave was complete. After 24 months, 26 of the 112 patients (23%) had lost their job. The presence of complete sick leave (OR 4.74, 95% CI 1.86-12.07) and the depression score of the hospital anxiety and depression scale (OR 1.18, 95% CI 1.02-1.36) were significantly and independently associated with job loss after 2 years follow-up. CONCLUSION: The occurrence of complete sick leave was found to be an independent risk factor for job loss in patients with chronic arthritis who have a disease related problem at work. PMID- 16710712 TI - Psychosocial work environment and burnout among emergency medical and nursing staff. AB - OBJECTIVES: The prevalence of burnout syndrome is increasing among doctors and nurses. The aim of this study was to analyse the relationship between the psychosocial work environment and burnout syndrome among emergency medical and nursing staff in Spain. A secondary aim was to determine if the effect of this psychosocial work environment on burnout was different for doctors and nurses. METHODS: A cross-sectional survey was carried out by means of a mail questionnaire among 945 emergency doctors and nursing staff of Spain. The outcome variable was three dimensions of burnout syndrome [emotional exhaustion (EE), personal accomplishment (PA), depersonalisation (DP)]. The explanatory variable was that psychosocial work environment evaluated according to Karasek and Johnson's demand-control model. The adjusted odds ratios (OR) and their 95% confidence intervals were calculated by logistical regression. RESULTS: The probability of high EE was greater among those exposed to high psychological demands, OR 4.66 (2.75-7.90), low job control, OR 1.65 (1.04-2.63), and low supervisors' social support, OR 1.64 (1.01-2.59). Emotional exhaustion dimension was negatively influenced by low control only among doctors. Those exposed to low job control had a higher risk of low PA, OR 2.55 (1.66-3.94). There was no evidence of negative effect of psychosocial risk factors on the DP. Prevalence of EE and PA was higher among doctors and nurses. CONCLUSIONS: The presence of risk factors derived from work organisation within the work place (psychosocial risk factors) increases the probability of presenting burnout syndrome and, above all, EE. PMID- 16710711 TI - The role of antioxidant supplementation in occupational exposure to waste anaesthetic gases. AB - OBJECTIVES: Although the genotoxicity related to waste anaesthetic gases is controversial, a consistent number of observations have provided evidence for an increased level of DNA strand breaks. The goal of the research was to investigate this hypothesis and estimate the genoprotective role of antioxidant supplementation in technical anaesthesiology staff working in operating theatres. METHODS: Heparinized venous blood samples were collected from 17 exposed technical anaesthesiology staff (mean age 34.3 +/- 3.5 years) and non-exposed control group (mean age 32.2 +/- 3.4 years) and examined in the alkaline comet assay for DNA strand breakage. Vitamin E (300 mg/day) plus vitamin C (500 mg/day) were supplemented to the technical anaesthesiology staff for 12 weeks and blood samples were retaken and evaluated by comet assay. RESULTS: The DNA breakage observed in the lymphocytes of the technical anaesthesiology staff was 21.5 +/- 5.0, as calculated by total comet score (TCS). This score was significantly higher (P<0.001) than in the controls (8.6 +/- 4.7) before antioxidant treatment. Supplementation of vitamins E plus C significantly (P<0.01) reduced the mean TCS as 14.2 +/- 6.1. CONCLUSION: The results of our study indicate that occupational exposure to anaesthetic gases induces oxidative DNA damage. Supplementation of the diet for 12 weeks with vitamin C and vitamin E resulted in a significant decrease in the DNA damage. PMID- 16710713 TI - The effect of recurrent involuntary job loss on the depressive symptoms of older US workers. AB - OBJECTIVES: The objective of this study was to assess whether recurrent involuntary job loss among US workers nearing retirement resulted in increasingly less severe changes in depressive symptoms with successive job losses. METHODS: With data drawn from the US Health and Retirement Survey (HRS), we used repeated measures longitudinal analysis to investigate the effect of recurrent job loss on follow-up depressive symptoms, measured up to 2 years following job loss. Study participants include 617 individuals, aged 51-61 years at the 1992 study baseline, who had at least one job loss between 1990 and 2000. Our primary outcome variable was a continuous measure of depressive symptoms, constructed from the 8-item Center for Epidemiologic Studies-Depression (CES-D) battery administered at every HRS wave. A second, dichotomous outcome, derived from the continuous measure, measured clinically relevant depressive symptoms. The exposure (recurrent job loss) was defined by binary dummy variables representing two and three/four job losses. All job losses were the result of either plant closing or layoff. RESULTS: Our main finding indicates that, after relevant covariates are controlled, compared to one job loss, two job losses result in a modest increase in the level depressive symptoms (not significant) at two-year follow-up. Three or more job losses result, on average, in a decline in depressive symptoms to a level near pre-displacement assessment (not significant). Somewhat in contrast, two job losses were found to be associated with increased risk of clinically relevant depressive symptoms. CONCLUSIONS: The principal finding confirms our hypothesis that, among US workers nearing retirement, repeated exposure to job separation results in diminished effects on mental health. Adaptation to the job loss stressor may underlie the observed response, although other explanations, including macroeconomic developments, are possible. PMID- 16710714 TI - Evaluation of factors influencing medication compliance in inpatient treatment of psychotic disorders. AB - RATIONALE: Short- and long-term compliance to prescribed antipsychotic drugs is of particular concern in regard to medication choice and treatment outcome in the care of psychotic disorders. OBJECTIVE: We evaluated patient-related and treatment-related factors associated with medication compliance in inpatients with a diagnosis of schizophrenia, schizoaffective disorder, or other psychotic disorder. METHODS: Within a naturalistic study in seven psychiatric hospitals, individuals with a psychotic disorder were assessed weekly on mental state, social functioning, side effects, and medication compliance. Logistic regression analyses were computed to assess patient and clinical predictors of medication compliance. RESULTS: We found a significant association between medication compliance and substance abuse (OR 0.52, CI 0.32-0.85), involuntary admission (OR 0.60, CI 0.41-0.89), history of aggressive behavior (OR 0.57, CI 0.38-0.85), and no school graduation (OR 0.59, CI 0.41-0.86). Individuals with pronounced paranoid or negative symptoms were also less compliant in taking their prescribed medication. There was no association between the initial inpatient antipsychotic medication regime and patients' compliance. Individuals who switched from a typical to an atypical antipsychotic drug were more compliant than those with their typical antipsychotic drug maintained. Those with higher medication compliance showed significantly greater improvement of their psychiatric symptoms during the inpatient stay. CONCLUSION: Patient-related in addition to disease related factors may strongly influence medication compliance. Besides more compliance with atypicals supposed by the literature, there may be a higher propensity for atypical drugs to be prescribed to those assumed to be more compliant. PMID- 16710716 TI - Idiopathic thrombocytopenic purpura in pregnancy: Analysis of 43 consecutive cases followed at a single Italian institution. PMID- 16710717 TI - HLA-B67 may be a male-specific HLA marker of susceptibility to relapsed childhood ALL in Hong Kong Chinese and HLA-A33 or HLA-B17 signifies a higher presentation leukocytosis: A retrospective analysis on 53 transplant candidates (1989-2003). AB - We performed a retrospective analysis on the human leukocyte antigen (HLA) data of 53 consecutive Chinese patients with high-risk childhood acute lymphoblastic leukemia (ALL) diagnosed from 1989 to 2003. A significantly higher frequency of HLA-B67 in the male relapse group of patients [OR, 23.08; 95% CI, 5.31-100.36; p = 0.0042; for statistical significance after Bonferroni correction (Bc) p (Bc) < 0.0083] was identified after Bonferroni correction. Although not surviving the Bonferroni correction, gender effects on the association were also observed with HLA-A11, HLA-A32, HLA-A33, and HLA-B22, which were however more prevalent in the female patients and particularly those developing relapse. Two patients with HLA A29 and HLA-B7 revealed significantly shortened survivals, suggestive of their potential prognostic impacts. Notably, for the first time, we found a significant correlation of leukocyte count with HLA types, where HLA-A33 (p = 0.006) or HLA B17 (p < 0.001) signifies higher leukocytosis at presentation. Taken together, our findings support the involvement of HLA in Chinese high-risk childhood ALL. PMID- 16710715 TI - The effect of acute tryptophan depletion on the BOLD response during performance monitoring and response inhibition in healthy male volunteers. AB - RATIONALE: Serotonin (5-HT) was implicated in both clinical and experimental studies in flexible, goal-directed behavior. However, the way in which 5-HT manipulations affect brain activation patterns underlying different subprocesses of cognitive flexibility remains largely unknown. OBJECTIVES: The aim of this study was to investigate the effect of a transient lowering of 5-HT on brain activation during performance monitoring and response inhibition. MATERIALS AND METHODS: We used acute tryptophan depletion (ATD), a well-known method to reduce central 5-HT, to investigate the effect of a transient lowering of 5-HT on the blood-oxygen-level dependent (BOLD) response in an event-related functional MRI study. Thirteen healthy male volunteers performed a modified Go/NoGo task in a counterbalanced, placebo-controlled, within-subject design. RESULTS: ATD significantly lowered plasma tryptophan but did not affect mood and cognitive performance. ATD decreased the BOLD response in the dorsomedial prefrontal cortex (BA 8) during performance monitoring. ATD did not affect the BOLD response during response inhibition. CONCLUSIONS: This study provides more evidence for the suggested role of 5-HT in performance monitoring. Because ATD studies have revealed inconsistent effects of ATD on performance and on brain activation, it was suggested that gender and personality traits are important variables to take into account for future research. PMID- 16710718 TI - Unusual presentation of mesenteric vasculitis as isolated dissection of the superior mesenteric artery. PMID- 16710719 TI - Human interleukin 24 (MDA-7/IL-24) protein kills breast cancer cells via the IL 20 receptor and is antagonized by IL-10. AB - The melanoma differentiation-associated gene-7 (mda-7/IL-24) is a unique member of the interleukin 10 (IL-10) family of cytokines, with ubiquitous tumor cell pro apoptotic activity. Recent data have shown that IL-24 is secreted as a glycosylated protein and functions as a pro-Th1 cytokine and as a potent anti angiogenic molecule. In this study, we analyzed the activity of Ad-mda7 and its protein product, secreted IL-24, against human breast cancer cells. We show that Ad-mda7 transduction of human breast cancer cells results in G(2)/M phase cell cycle arrest and apoptotic cell death, which correlates with secretion of IL-24 protein. Neutralizing antibody against IL-24 significantly inhibited Ad-mda7 cytotoxicity. IL-24 and IL-10 both engage their cognate receptors on breast cancer cells resulting in phosphorylation and activation of STAT3, however, IL-10 receptor binding failed to induce cell killing, indicating that tumor cell killing by IL-24 is independent of STAT3 phosphorylation. Treatment with exogenous IL-24 induced apoptosis in breast cancer cells and this effect was abolished by addition of anti-IL-24 antibody or anti-IL-20R1, indicating that bystander cell killing is mediated via IL-24 binding to the IL-20R1/IL-20R2 heterodimeric receptor complex. Co-administration of the related cytokine IL-10 inhibited killing mediated by IL-24 and concomitantly inhibited IL-24 mediated up regulation of the tumor suppressor proteins, p53 and p27(Kip1). In summary, we have defined a tumor-selective cytotoxic bystander role for secreted IL-24 protein and identified a novel receptor-mediated death pathway in breast cancer cells, wherein the related cytokines IL-24 and IL-10 exhibit antagonistic activity. PMID- 16710720 TI - Sequence effects in a spatial cueing task: endogenous orienting is sensitive to orienting in the preceding trial. AB - In a spatial cueing paradigm it was investigated whether endogenous orienting is sensitive to orienting processes in the previous trial. Specifically, the effect of the previous cue direction, the previous trial type (valid, invalid, neutral, catch) and target alternation effects were studied. Strategic effects were shown as attentional costs and benefits were larger after a valid than after an invalid trial. Following catch trials, an overall response slowing was observed, but costs and benefits were unaffected. This was interpreted as a reduction in alertness and as support for the dissociation between spatial and temporal attentional mechanisms. Repetition of target position per se had no effect, but in neutral trials responses were slower to targets appearing at the location that was cued in the previous trial, independent of validity of the preceding trial. This suggests that long-term inhibition-of-return can occur between trials when attention is controlled endogenously. PMID- 16710721 TI - [Minimally invasive approach to implantation of a hip endoprosthesis]. AB - Early experiences with the two-incision approach at the hip joint are reported. We started in July 2002 and until now 272 procedures have been performed. Blood loss (292 ml) and the need for rehabilitation are reduced, the patients will achieve a Harris Hip Score of more than 90 points within 6 or 8 weeks. Complications related to the approach primarily include irritation of the lateral femoral cutaneous nerve. During the learning curve when the initial shaft model was used, shaft fractures or fissures occurred in 4% of the cases. This complication was no longer observed with the ML Taper shaft model now employed (n=102). The development and establishment of minimally invasive approaches constitute a valuable and seminal option for implantation of hip endoprostheses. In the medium term, the time spent in rehabilitation could conceivably be reduced. Conventional approaches still represent the gold standard. PMID- 16710722 TI - Intra-osseous pseudoaneurysm following curettage of an aneurysmal bone cyst. AB - BACKGROUND: Pseudoaneurysms secondary to bone tumours are rare and most of the reported cases are related to osteochondromas, either due to direct pressure or following surgery. Aneurysmal bone cysts are relatively common bony lesions usually treated by curettage. DISCUSSION: We describe an unusual case of pseudoaneurysm of the anterior tibial artery complicating curettage of an aneurysmal bone cyst which presented as a rapidly enlarging mass clinically thought to be rapid recurrence of the tumour. This was successfully treated by embolisation. PMID- 16710723 TI - The AAPC case, with an early onset of colorectal cancer. AB - INTRODUCTION: Attenuated adenomatous polyposis coli (AAPC) is a variant of the familial adenomatous polyposis (FAP) characterized by the occurrence of sparse polyps in the colon, stomach, and duodenum with a late onset of colorectal cancer. The AAPC syndrome is associated with mutations at the 5' region of the APC gene. Until recently, the fragment encompassing codons 157 and 170 was considered as boundary for the described cases of AAPC and FAP syndromes. MATERIALS AND METHODS: This study describes a case of the AAPC syndrome caused by a CCTT deletion at codon 173, with polyps diagnosed at the age of 17. The father and grandfather of the proband died of colorectal cancer (CRC), which developed from untreated polyps, at the age 35 and 40, respectively. RESULTS AND DISCUSSIONS: In the case of the proband's father, the untreated polyps led to death after 12 years. The proband revealed a low number of polyps and an extra colon feature characteristic of AAPC, but the polyps onset and the death of CRC of two family members, who refused colectomy, was very early and characteristic for FAP. An atypical course of AAPC must be taken into consideration both in genetic counseling and in qualifying the patients with AAPC for the surgical treatment. PMID- 16710727 TI - [Social anxiety disorders]. AB - A main feature of social anxiety disorders is the fear or avoidance of social situations in which one is the center of attention and fears criticism or negative judgement. According to current diagnostic systems, three types of these disorders can be classified: specific social phobia, generalised social phobia, and avoidant personality disorder. Social anxiety disorders are prevalent mental disorders (lifetime prevalence up to 13%). Other mental disorders often appear together with them. Following the criteria for evidence-based medicine, psychotherapeutic and pharmacologic interventions are described. Regarding psychotherapeutic approaches, the best results have been seen with cognitive behavioural methods. Among psychopharmacologic treatments, selective serotonin reuptake inhibitors and the selective noradrenalin reuptake inhibitor Venlafaxin are the first-choice treatments. PMID- 16710729 TI - Accessory lateral discoid meniscus. AB - The lateral meniscus tends to have more developmental variation than the medial counterpart. This is a report of an accessory discoid layer of lateral meniscus. All arthroscopic, magnetic resonance imaging and histopathological views are presented. PMID- 16710728 TI - Arthroscopic treatment of patients with moderate arthrofibrosis after total knee replacement. AB - The purpose of this study was to document the effect of arthroscopic management in patients with knee stiffness after total knee replacement. We present a case series study, in which 32 patients have been treated for moderate arthrofibrosis of the knee after total knee replacement, with the same regimen. We have excluded all cases of stiffness, because of infection, mechanical mal-alignment, loosening of the implants and other obvious reasons of stiffness of the knee, rather than pure arthrofibrosis. All patients first underwent a trial of conservative treatment before going for arthroscopic management. A pain catheter for femoral nerve block was inserted just before anesthesia for post-operative pain management. Arthroscopic arthrolysis of the intra-articular pathology was performed in a standardized technique with release of all fibrous bands in the suprapatellar pouch, reestablishing the medial and lateral gutter, release of the patella, resection of the remaining meniscal tissue or an anterior cyclops, if needed. Intensive physiotherapy and continuous passive motion were to start immediately post-operatively. All the patients were available for the follow up and they were evaluated using the knee society rating system. A total of 25 of the 32 procedures resulted in an improvement of the patients knee score. All the knees operated upon had intra-articular fibrous bands, hypertrophic synovitis and peri-patellar adhesions. A total of eight patients suffered from an anterior cyclops lesion and six patients showed pseudomenicus. In 19 cases a medial and lateral relapse of the patella was performed; only 5 patients got an isolated lateral release. The mean knee flexion was 119 degrees (100-130) at the end of arthroscopy and was 97 degrees (75-115) at the last follow up. The eight patients with extension lags decreased from 27 degrees (10 degrees-35 degrees) pre operatively to 4 degrees (0-10) at time of follow up. The average knee society ratings increased from 70 points prior to the arthroscopy to 86 at time of follow up, which was found to be statistically significant (P < 0.01, student's t test). The average function score also showed improvement from 68 points pre-operatively to 85 at the time of final follow up. The average pain scores improved from 30 points pre-operatively to 41 at the time of final follow up. Our results showed that arthroscopic management of knee stiffness following total knee replacement is a safe and efficient method of treatment. Pain and functional knee scores can improve markedly. PMID- 16710730 TI - Rapid chondrolysis after arthroscopic partial lateral meniscectomy in athletes: a case report. AB - We present a patient with a severe chondrolysis after arthroscopic partial lateral meniscectomy in a 17-year-old high school basketball player. This is a rare but severe complication after arthroscopic partial lateral meniscectomy. At 7 months after the first operation, a second-look arthroscopy showed numerous cartilaginous debris floating in the knee and a high-grade cartilage damage on the lateral compartment of the tibia. This unexpected complication and a consideration of its etiology are shown. PMID- 16710732 TI - Theoretical study of the intermolecular H-bonding and intermolecular proton transfer between isocytosine tautomeric forms and R,S-lactic acid. AB - Eight H-bonded complexes between isocytosine (isoC) tautomeric forms and R/S lactic acid (LA) have been studied at the B3LYP and HF levels of theory using 6 31+G(d) basis set. The energy barriers of the intermolecular proton transfers were also estimated as the results showed that they are several times lower than those of the intramolecular proton transfers of isoC in the gas phase. Furthermore, the energy barriers of the tautomerizations in which the carboxylic H-atom takes part are several times lower than those in which the LA OH group assists the proton transfer. PMID- 16710731 TI - Soft tissue graft interference fit fixation: observations on graft insertion site healing and tunnel remodeling 2 years after ACL reconstruction in sheep. AB - Using soft tissue grafts for anterior cruciate ligament (ACL) reconstruction, insertion site healing plays a crucial role in the long-term fate of the graft. It has been shown in an experimental animal study that using a soft tissue graft and anatomic graft fixation, a direct ligamentous insertion alike the native ACL developed 24 weeks postoperatively. Yet there are no reports on the long-term insertion site healing of anatomically fixed soft tissue grafts. The objective of this study was to evaluate graft insertion site healing, the intra-tunnel fate of the graft and its osseous replacement 2 years after ACL reconstruction in sheep. The left ACLs of six sheep were replaced by an autologous flexor tendon split graft and anatomically fixed with biodegradable poly-(D, L-lactide) interference screws. Animals received polychromic sequential labeling at different points in time to determine bone apposition per period. For evaluation of the insertion site healing and intra-tunnel changes, MRI scans were taken in vivo. Following sacrifice, radiographic imaging, conventional histology and fluorescence microscopy was undertaken. Most of the specimens showed a wide direct ligamentous insertion. It showed patterns alike the direct ligament insertion seen in intact ACLs. The intra-tunnel part of the graft had completely lost its tendon-like structure and in two cases, it was separated from the graft insertion by a thick bony layer. The biodegradable interference screw was fully degraded in all specimens. Ossification of the former drill tunnels was intense, showing only partial-length tunnel remnants in one femoral and three tibial specimens. As the graft heals to the joint surface and the aperture site is closed with soft tissue, mechanical stress of the intra-tunnel part of the graft is eliminated and the bone tunnel is protected from synovial fluid, resulting in osseous bridging of the tunnel aperture site, accelerated intra-tunnel graft resorption and its osseous replacement. PMID- 16710734 TI - Gentamicin negatively influenced osteogenic function in vitro. AB - Local delivery of gentamicin is an accepted method of infection prophylaxis in the surgery of open fractures. However, the few reports of studies into the effect of locally applied gentamicin on osteoblasts used inadequate methods. In our study, we used the well-characterised C2C12 cell line with reproducible differentiation pathway into the osteoblast lineage. We investigated the viability, cell number, alkaline phosphatase activity, and the expression of osteogenic genes of C2C12 cells after exposure to gentamicin at concentrations of 12.5-800 microg/ml for 48 h. Exposure of C2C12 cells to gentamicin (12.5-800 mg/ml) for 48 h showed no significant changes in the cell number, but cell viability was decreased by one-third at the tested concentrations of 200-800 microg/ml. The alkaline phosphatase activity was significantly decreased by one third to one-half at any tested concentration (12.5-800 microg/ml) of gentamicin. Any tested concentration of gentamicin up to 800 microg/ml for 48 h did not inhibit or decrease the osteogenic gene expression of osterix and alkaline phosphatase of the C2C12 cells. In conclusion, gentamicin at high concentrations as achieved by local application reduced cellular viability and alkaline phosphatase activity in vitro and therefore may be detrimental for bone healing and repair in vivo. PMID- 16710733 TI - Protein oxidation in obesity and insulin resistance. AB - INTRODUCTION: Advanced oxidation protein products (AOPP) are considered reliable markers to estimate the degree of oxidant-mediated protein damage. Data on oxidative stress in childhood obesity and insulin resistance are limited. OBJECTIVE: The aim of this study was to investigate the AOPP level as an oxidative stress marker in obesity and insulin resistance. The study included 57 pubertal obese children and adolescents (30 girls and 27 boys) and 20 healthy pubertal children and adolescents (11 girls and 9 boys). MATERIALS AND METHODS: All participants in the obesity group underwent an oral glucose tolerance test (OGTT) and two separate groups were formed according to the existence of insulin resistance. RESULTS: AOPP levels were measured in the obesity and control groups spectrophotometrically. The obesity group consisted of 25 children and adolescents with insulin resistance and 32 subjects without insulin resistance. AOPP levels in the obesity group were found to be significantly higher than those in the control group. Although AOPP levels in the subjects with insulin resistance were higher than the subjects without insulin resistance, there was no significant difference between AOPP levels of subgroups with insulin resistance and without insulin resistance. CONCLUSION: This study showed protein oxidation in obesity with a novel oxidative stress marker and it also suggests that insulin resistance may play an important role as a source of oxidative stress in the development of other diseases after pubertal years. PMID- 16710736 TI - Population study and evaluation of 20 Y-chromosome STR loci in Germans. AB - The nine European minimal haplotype (EMH) loci, the two SWGDAM loci and five further single-copy Y-chromosomal short tandem repeats (Y-STRs) DYS446, DYS447, DYS448, DYS449, DYS463, and the multicopy loci DYS464 were evaluated in the German population groups Dresden, Hamburg, Rostock, Munich, and the Sorbs who are a Slavic-speaking minority in Lusatia. Highest gene diversities in all populations were shown for DYS464, DYS385, and DYS449 (D=0.8559-0.9486). The haplotype diversity for the European minimal haplotype loci ranged between 0.9852 for Sorbs and 0.9983 for the Hamburg population showing that there is a significant portion of haplotypes which could not be resolved. Advanced typing using DYS446, DYS447, DYS448, DYS449, DYS463, and DYS464 discriminated all non related individuals of the Dresden, Hamburg, and Rostock populations. Evaluation of the Y-STRs was accomplished by sequence analysis of all allelic fragments of the allelic ladders and microvariant alleles of DYS385 and the determination of the amounts of stutter products of the loci. PMID- 16710737 TI - Depth-dependent compressive equilibrium properties of articular cartilage explained by its composition. AB - For this study, we hypothesized that the depth-dependent compressive equilibrium properties of articular cartilage are the inherent consequence of its depth dependent composition, and not the result of depth-dependent material properties. To test this hypothesis, our recently developed fibril-reinforced poroviscoelastic swelling model was expanded to include the influence of intra- and extra-fibrillar water content, and the influence of the solid fraction on the compressive properties of the tissue. With this model, the depth-dependent compressive equilibrium properties of articular cartilage were determined, and compared with experimental data from the literature. The typical depth-dependent behavior of articular cartilage was predicted by this model. The effective aggregate modulus was highly strain-dependent. It decreased with increasing strain for low strains, and increases with increasing strain for high strains. This effect was more pronounced with increasing distance from the articular surface. The main insight from this study is that the depth-dependent material behavior of articular cartilage can be obtained from its depth-dependent composition only. This eliminates the need for the assumption that the material properties of the different constituents themselves vary with depth. Such insights are important for understanding cartilage mechanical behavior, cartilage damage mechanisms and tissue engineering studies. PMID- 16710735 TI - Mast cell tryptase in postmortem serum-reference values and confounders. AB - We have investigated the effects of some factors suspected of inducing spuriously increased tryptase concentrations, specifically sampling site, conjunctival petechial bleeding and prone position at the time of death as indicators of premortem asphyxia, and resuscitation efforts by external cardiac massage. Tryptase was measured in blood from the femoral vein in 60 deaths: 39 control cases who died rapidly (within minutes) from natural causes (sudden cardiac death and acute aortic dissection), 16 with death caused by prolonged asphyxia (traumatic compression of the chest and suffocation due to body position or smothering), and five anaphylactic deaths. In 44 of these cases, tryptase was measured in both heart and femoral blood. Mast cell tryptase was analyzed with a commercial FEIA method (Pharmacia Diagnostics AB, Uppsala, Sweden) measuring both alpha- and beta-tryptase. Assuming that tryptase values in the control group were gamma distributed, we calculated the upper normal limits for tryptase concentrations in femoral blood. It was found that 95% of the controls had values below 44.3 mug/l (femoral blood), SD 5.27 mug/l. All but one of the anaphylactic deaths had tryptase concentrations exceeding that limit. Tryptase was significantly elevated in femoral blood from anaphylactic deaths (p<0.007), compared with the controls. Also, in the cases where death had occurred due to asphyxia tryptase was elevated in femoral blood (p<0.04). A significant difference in tryptase concentrations was seen between blood from the heart and the femoral vessels (p<0.02) in the whole material (n=44). Tryptase concentrations in femoral blood were not influenced by prone position at death, or resuscitation efforts. It is concluded that asphyxia premortem seems to affect tryptase concentrations, that postmortem tryptase measurements should be done in serum from femoral blood, and that the normal upper limit, covering 95%, is 44.3 microg/l. PMID- 16710738 TI - In memoriam Hannes Schuepp 1931-2006. PMID- 16710740 TI - Scleroderma-like remodeling induced by type V collagen. AB - Recently, we discovered that New Zealand rabbits immunized with human type V collagen plus Freund's adjuvant present fibrosis and vasculitis of organs usually affected by systemic sclerosis. In this way, we studied the fibrillogenesis process to identify possible factors involved in altered remodeling observed in this scleroderma-like model. Additionally, we have done a very preliminary comparison with human skins obtained from scleroderma patients (n=3). Female New Zealand rabbits (n=10) were immunized subcutaneously with two doses of 1 mg collagen V (COL V) plus complete Freund's adjuvant for a 30-day interval, followed by two additional intramuscular booster immunizations in incomplete Freund's adjuvant for a 15-day interval. Animals from control group (n=10), were only inoculated with complete and incomplete Freund's adjuvant given at same conditions of COL V. Histological analysis of skins from animals and patients were done by Masson's trichrome staining, and immunofluorescence method to detect collagen fibers and interactions of types I, III and V collagen in the remodeling process. The analysis of animal skins showed collagen fibril deposits in the dermis after 7 days of sensibilization and an increase in these deposits after 75 and 120 days, respectively. Skin thickness and atrophy of sebaceous and sweat glands were progressively more intense in late sacrificed animals and correlated with increased amount of collagen deposition. Surprisingly, type V collagen was overexpressed both in animals and patients, forming dense and atypical collagen fibers in the dermis. We suggest that this anomalous expression of morphologically different type V collagen could justify the remodeling observed in scleroderma plaque. PMID- 16710741 TI - Recombined DNA vaccines encoding calreticulin linked to HPV6bE7 enhance immune response and inhibit angiogenic activity in B16 melanoma mouse model expressing HPV 6bE7 antigen. AB - Calreticulin (CRT) has been reported to have an effect of upregulating MHC class I presentation as well as inhibiting angiogenesis in vitro and in vivo. Combination of dual mechanisms of enhanced immunogenicity of human papillomavirus (HPV) 6bE7 antigen and antiangiogenesis may be introduced in the strategy of vaccines against condyloma acuminatum (CA) resulting from HPV infection. Therefore, we constructed DNA vaccines by employing different lengths of CRT chimerically linked to a model antigen HPV6bE7 and investigated the immunological and antiangiogenic effects of these vaccines in a B16 melanoma model that express HPV6bE7 antigen. Our results showed that vaccination with CRT180/HPV6bE7 or CRT120/HPV6bE7 enhanced the presence of CD8(+) T cells and TCRgammadelta T cells in vivo, increased the specific lysis activity against E7-expressing cells and secretion levels of IL-2 and IFN-gamma by activating T cells in vitro significantly. Moreover, recombined CRT180 or CRT120 with HPV6bE7 vaccines could elicit a more efficient E7-specific immune response than HPV6bE7 alone. The similarity of immunological enhancement of CRT180/HPV6bE7 and CRT120/HPV6bE7 implies that the immunologically active region mainly exist in fragment 1-120 aa. Furthermore, CRT180/HPV6bE7 and CRT180 displayed remarkable superiority over CRT120/HPV6bE7 in vivo angiogenesis assay, suggesting that the antiangiogenic activity of CRT resides in a domain between aa 120 and 180. Vaccination with CRT180/HPV6bE7 generated the best protective effect of delaying tumor formation and reduction of tumor size in tumor growth inhibition experiment among all DNA constructs. Therefore, CRT180/HPV6bE7 vaccine may enhance the immunological response to HPV6bE7 and inhibit angiogenesis. This construct may be useful in preventing HPV-associated dermatosis and may be developed as a promising strategy to control CA. PMID- 16710742 TI - Expression of lysosome-associated membrane protein 1 (Lamp-1) and galectins in human keratinocytes is regulated by differentiation. AB - Lysosomes and their components are suspected to be involved in epidermal differentiation. In this study, lysosomal enzyme activities, expression of the lysosome-associated membrane protein 1 (Lamp-1) and expression of the epidermal galectins-1, -3 and -7 were investigated in human keratinocytes cultured at different cell densities (subconfluence, confluence and postconfluence) in order to induce differentiation. Detected by Western blot and immunofluorescence, Lamp 1 expression is transiently upregulated at culture confluence, but reduced at postconfluence. Northern blot analyses performed on subconfluent, confluent and post-confluent cultures of keratinocytes show that Lamp-1 mRNA expression is also upregulated at culture confluence, but downregulated at postconfluence. Measurements of lysosomal enzyme activities indicate a transient upregulation at culture confluence, whereas cathepsins B, C and L are particularly downregulated at postconfluence. Cell density and differentiation of epidermal cells also differentially regulates galectin expression in autocrine cultures. As the expression of galectin-1 mRNA is high in subconfluent cells, it is assumed to be associated with their proliferative state. On the other hand, as the mRNA levels for galectins-3 and -7 are notably upregulated at culture confluence (galectin-7) or at postconfluence (galectin-3), their expression is thought to be related to the differentiated state of keratinocytes. However, we collected evidence by confocal microscopy that galectin-3 and Lamp-1 do not colocalize in vitro in keratinocytes. Altogether, our results suggest that the upregulated Lamp-1 expression at confluence could be involved in keratinocyte differentiation, but apparently not through interaction with galectin-3. PMID- 16710743 TI - Effects of various oxidants and antioxidants on the p38-MAPK signalling pathway in the perfused amphibian heart. AB - We investigated the effects of different antioxidants such as L-ascorbic acid, catalase, and superoxide dismutase (SOD), on the p38-MAPK activation induced by oxidative stress in the isolated perfused amphibian heart. Oxidative stress was exemplified by perfusing hearts with 30 microM H(2)O(2) for 5 min or with the enzymatic system of xanthine/xanthine oxidase (200 microM/10 mU/ml, respectively) for 10 min. H(2)O(2)-induced activation of p38-MAPK (7.04 +/- 0.20-fold relative to control values) was totally attenuated by L-ascorbic acid (100 microM) or catalase (150 U/ml). These results were confirmed by immunohistochemical studies in which the phosphorylated form of p38-MAPK was localised in the perinuclear region and dispersedly in the cytoplasm of the ventricular cells during H(2)O(2) treatment, a pattern that was abolished by catalase or L-ascorbic acid. p38-MAPK was also activated (2.34 +/- 0.17-fold) by perfusing amphibian hearts with the reactive oxygen species (ROS)-generating system of xanthine/xanthine oxidase and this activation sustained in the presence of 150 U/ml catalase (2.16 +/- 0.26 fold), 50 U/ml SOD (2.02 +/- 0.07) or 100 microM L-ascorbic acid (2.18 +/- 0.10), but was suppressed by the combination of 150 U/ml catalase and 50 U/ml SOD. Finally, our studies showed that xanthine/xanthine oxidase induced the phosphorylation of the potent p38-MAPK substrates MAPKAPK2 (3.14 +/- 0.27-fold) and HSP27 (5.32 +/- 0.83-fold), which are implicated in cell protection, and this activation was reduced by the simultaneous use of catalase and SOD. PMID- 16710745 TI - Activity of lysosomal exoglycosidases in human gliomas. AB - There is a lot of data suggesting that modifications of cell glycoconjugates may be important in progression of cancer. In the present work we studied activities of lysosomal exoglycosidases: beta-hexosaminidase and its isoenzymes A and B, beta-galactosidase and alpha-mannosidase, in human gliomas. Enzyme activity was determined spectrophotometrically based on the release of p-nitrophenol from p nitrophenyl-derivative of appropriate sugars. The activities of the exoglycosidases tested were significantly higher in malignant glial tumors than in control tissue (normal brain tissue) and non-glial tumors. The highest activities of exoglycosidases were observed in high-grade gliomas, and a positive correlation of enzyme activities and degree of malignancy was noted. Our results suggest that lysosomal exoglycosidases may participate in the progression and dynamical development of glial tumors. PMID- 16710746 TI - 1p/19q chromosome deletions in metastatic oligodendroglioma. AB - Extracranial metastasis of primary brain tumors is a rare phenomenon. Of the few cases reported of metastatic oligodendroglioma, only two have evaluated genetic alterations, specifically deletions of chromosomes 1p and 19q. Herein, we report two additional patients with metastatic anaplastic oligodendroglioma to bone, both followed until death. All available pathology specimens were reviewed and genetic analysis was performed in one of the cases. Although the bone metastasis was non-informative, the primary intracranial tumor revealed codeletions of the 1p and 19q chromosomal arms, commonly recognized as the genetically favorable profile of oligodendrogliomas. Both patients died of complications related to their systemic disease and did not have any radiologic evidence of intracranial progression at the time of their last MRI studies. Along with the reported literature, our data suggest that despite their generally favorable behavioral profiles, oligodendroglial tumors with 1p/19q deletions may be more prone to metastasis as they progress. Genetic analysis serves a valuable ancillary role in the diagnostic workup of such cases. PMID- 16710744 TI - Prolonged AMPK activation increases the expression of fatty acid transporters in cardiac myocytes and perfused hearts. AB - Recently, fatty acid transport across the plasma membrane has been shown to be a key process that contributes to the regulation of fatty acid metabolism in the heart. Since AMP kinase activation by 5-aminoimidazole-4-carboxamide-1-beta-D: ribofuranoside (AICAR) stimulates fatty acid oxidation, as well as the expression of selected proteins involved with energy provision, we examined (a) whether AICAR induced the expression of the fatty acid transporters FABPpm and FAT/CD36 in cardiac myocytes and in perfused hearts and (b) the signaling pathway involved. Incubation of cardiac myocytes with AICAR increased the protein expression of the fatty acid transporter FABPpm after 90 min (+27%, P < 0.05) and this protein remained stably overexpressed until 180 min. Similarly, FAT/CD36 protein expression was increased after 60 min (+38%, P < 0.05) and remained overexpressed thereafter. Protein overexpression, which occurred via transcriptional mechanisms, was dependent on the AICAR concentration, with optimal induction occurring at AICAR concentrations 1-5 mM for FABPpm and at 2-8 mM for FAT/CD36. The AICAR (2 h, 2 mM AICAR) effects on FABPpm and FAT/CD36 protein expression were similar in perfused hearts and in cardiac myocytes. AICAR also induced the plasmalemmal content of FAT/CD36 (+49%) and FABPpm (+42%) (P < 0.05). This was accompanied by a marked increase in the rate of palmitate transport (2.5 fold) into giant sarcolemmal vesicles, as well as by increased rates of palmitate oxidation in cardiac myocytes. When the AICAR-induced AMPK phosphorylation was blocked, neither FAT/CD36 nor FABPpm were overexpressed, nor were palmitate uptake and oxidation increased. This study has revealed that AMPK activation stimulates the protein expression of both fatty acid transporters, FAT/CD36 and FABPpm in (a) time- and (b) dose-dependent manner via (c) the AMPK signaling pathway. AICAR also (d) increased the plasmalemmal content of FAT/CD36 and FABPm, thereby (e) increasing the rates of fatty acid transport. Thus, activation of AMPK is a key mechanism regulating the expression as well as the plasmalemmal localization of fatty acid transporters. PMID- 16710747 TI - Radiation induced adult medulloblastoma: a case report. AB - Adult medulloblastoma is a rare intracranial tumor. Our patient is a 61 year old woman treated with cranial irradiation 15 years previously for a low grade astrocytoma in the left posterior temporal lobe that was recently diagnosed with medulloblastoma in the right cerebellum. This is the first reported case of radiation induced adult medulloblastoma. PMID- 16710748 TI - A phase II study of carboplatin and chronic high-dose tamoxifen in patients with recurrent malignant glioma. AB - PURPOSE: To determine the response rate, time to disease progression, survival, and toxicity of intravenous carboplatin and chronic oral high-dose tamoxifen in patients with recurrent malignant gliomas. PATIENTS AND METHODS: Patients with histological confirmation of recurrent malignant gliomas were eligible for this multicenter phase II trial. Treatment consisted of 400 mg/m2 carboplatin intravenously every 4 weeks and oral high dose chronic tamoxifen (80 mg bid in women and 100 mg bid in men). RESULTS: Twenty seven patients met the eligibility criteria and were evaluable for response. The histological subtypes were: 16 (59%) glioblastoma multiforme (GBM), malignant astrocytoma (5 patients), malignant mixed glioma (5 patients), and glioblastoma/gliosarcoma (1 patient). Twenty-two patients (82%) had an ECOG performance status of 0 or 1. No complete responses were observed, 4 patients (15%) achieved a partial response, and 14 patients (52%) had stable disease. Median time to progression was 3.65 months (95%CI 2.56, 4.83). Median overall survival was 14.09 months (95%CI 7.06, 19.91). One patient with a recurrent GBM had a sustained partial response and is progression free 81 months since starting treatment. Another patient with mixed malignant oligoastrocytoma also had a prolonged partial response (lasting 63 months) and is alive 84 months after treatment for recurrence. The most frequently reported grade 3 or 4 toxicities were fatigue (19%), nausea (11%) and anorexia (11%). CONCLUSIONS: Carboplatin and high dose tamoxifen has similar response rates to other regimens for recurrent malignant gliomas and are probably equivalent to those found using tamoxifen as monotherapy. Long-lasting periods of disease free survival in some patients (particularly those with malignant mixed oligo astrocytomas) were found. PMID- 16710750 TI - Endometrial preparation for in vitro oocyte maturation: early use of estrogen increases endometrial tissue and requires lower daily dosage: a cross over trial in 'mock' cycles. AB - PURPOSE: Determine if estrogen used since the beginning of the menstrual flow could improve endometrial tissue compared to standard endometrial preparation for in vitro maturation cycles. METHODS: Twenty polycystic ovary syndrome women were submitted to two estrogen therapy schedules: standard schedule; estrogen began on the day of planned egg retrieval (dosage was based on endometrial thickness); and long schedule; estrogen began on the first day of menstruation. No oocyte retrieval or embryo transfer was performed. Three-dimensional ultrasound was performed on the day of planned egg retrieval and one week later for endometrial evaluation. RESULTS: A higher endometrial thickness and volume was found in long schedule on both evaluations. The number of patients that have used 10 mg/day of estradiol was significant higher in the standard schedule (65%x0%). No other significant difference was found. CONCLUSIONS: The early use of estrogen improves endometrial tissue and requires lower daily dosage. PMID- 16710754 TI - Pathway of oxidative folding of a 3-disulfide alpha-lactalbumin may resemble either BPTI model or hirudin model. AB - Pathways of oxidative folding of disulfide proteins display a high degree of diversity and vary among two extreme models. The BPTI model is defined by limited species of folding intermediates adopting mainly native disulfide bonds. The hirudin model is characterized by highly heterogeneous folding intermediates containing mostly non-native disulfide bonds. alphaLA-IIIA is a 3-disulfide variant of alpha-lactalbumin (alphaLA) with a 3-D conformation essentially identical to that of intact alphaLA. alphaLA-IIIA contains 3 native disulfide bonds of alphaLA, two of them are located at the calcium binding beta-subdomain (Cys61-Cys77 and Cys73-Cys91) and the third bridge is located within the alpha helical domain of the molecule (Cys28-Cys111). We investigate here the pathway of oxidative folding of fully reduced alphaLA-IIIA with and without stabilization of its beta-subdomain by calcium binding. In the absence of calcium, the folding pathway of alphaLA-IIIA was shown to resemble that of hirudin model. Upon stabilization of beta-sheet domain by calcium binding, the folding pathway of alphaLA-IIIA exhibits a striking similarity to that of BPTI model. Three predominant folding intermediates of alphaLA-IIIA containing exclusively native disulfide bonds were isolated and structurally characterized. Our results further demonstrate that stabilization of subdomains in a protein may dictate its folding pathway and represent a major cause for the existing diversity in the folding pathways of the disulfide-containing proteins. PMID- 16710755 TI - Directing DNA methylation to inhibit gene expression. AB - : 1. DNA methylation is a critical epigenetic modification that silences gene transcription, participates in X-chromosome inactivation in females, and regulates genomic imprinting. 2. We have devised a method to inhibit transcriptional initiation by constructing short methylated oligonucleotides which induce DNA methylation at specific loci. 3. The methodology by which we devise these oligonucleotides is described, using oligonucleotides directed against the oncogene, Bcl-2.4. The human Bcl-2 gene contains two promoters, each of which contains a CpG island in its core region. Oligonucleotides are designed which can inhibit Bcl-2 transcription and lead to decreased mRNA and protein in vitro. When compared to standard anti-sense oligonucleotide action, these methylated oligonucleotides are far more sensitive and potentially, longer acting. 5. In principle, using this methodology, it should be possible to design methylated oligonucleotides that can methylate CpG islands and thereby downregulate any gene. PMID- 16710756 TI - Identification of endophilins 1 and 3 as selective binding partners for VGLUT1 and their co-localization in neocortical glutamatergic synapses: implications for vesicular glutamate transporter trafficking and excitatory vesicle formation. AB - 1. Selective protein-protein interactions between neurotransmitter transporters and their synaptic targets play important roles in regulating chemical neurotransmission. We screened a yeast two-hybrid library with bait containing the C-terminal amino acids of VGLUT1 and obtained clones that encode endophilin 1 and endophilin 3, proteins considered to play an integral role in glutamatergic vesicle formation. 2. Using a modified yeast plasmid vector to enable more cost effective screens, we analyzed the selectivity and specificity of this interaction. Endophilins 1 and 3 selectively recognize only VGLUT1 as the C terminus of VGLUT2 and VGLUT3 do not interact with either endophilin isoform. We mutagenized four conserved stretches of primary sequence in VGLUT1 that includes two polyproline motifs (Pro1, PPAPPP, and Pro2, PPRPPPP), found only in VGLUT1, and two conserved stretches (SEEK, SYGAT), found also in VGLUT2 and VGLUT3. The absence of the VGLUT conserved regions does not affect VGLUT1-endophilin association. Of the two polyproline stretches, only one (Pro2) is required for binding specificity to both endophilin 1 and endophilin 3. 3. We also show that endophilin 1 and endophilin 3 co-localize with VGLUT1 in synaptic terminals of differentiated rat neocortical neurons in primary culture. These results indicate that VGLUT1 and both endophilins are enriched in a class of excitatory synaptic terminals in cortical neurons and there, may interact to play an important role affecting the vesicular sequestration and synaptic release of glutamate. PMID- 16710757 TI - Biochemical and clinical improvement of cytotoxic state by amantadine sulphate. AB - 1. The main idea of the open clinical trial was to compare the income and outcome clinical picture and the evolution of the biochemical markers in the defined intervals in closed head injury group patients. 2. In the group of 32 patients, mean age 40.78+/-15.36 years suffering from closed traumatic brain injury the following markers were measured: glycaemia, malondialdehyde (MDA) as marker of lipid peroxidation, beta-caroten, total SH groups as marker of protein oxidation in the following intervals: between the 1st and the 3rd, between the 3rd and the 7th, between the 1st and the 7th day respectively. 3. Glycaemia significantly decreased since the 1st day till the 3rd day (p < 0.05) and since the 1st day till the 7th day (p < 0.05) but it was not significantly changed since the 3rd day till the 7th day (p > 0.05). 4. MDA 1st x MDA 3rd p > 0.05 insignificant change, MDA 1st x MDA 7th p < 0.001--high significant decrease, MDA 3rd x MDA 7th -p < 0.0001--very high significant decrease. 5. Beta-caroten the 1st x beta caroten the 3rd day was insignificantly changed-p > 0.05, the 3rd x the 7th day beta-caroten increased significantly--p < 0.0002, the 1st day x 7th day beta caroten significantly increased--p < 0.0001. 6. We examined the SH groups only in nine patients, due to technical problems and SH groups decrease on the 3rd day (p < 0.005). 7. In 18 amantadine sulphate subgroups (randomly selected), there was 5.5% lethality and mean outcome GCS (outGCS) 9.83+/-3.8, while lethality of the control subgroup (n=14) was 42.9%, mean outGCS 6.28+/-3.5. PMID- 16710758 TI - Common human 5' dopamine transporter (SLC6A3) haplotypes yield varying expression levels in vivo. AB - 1. Individuals display significant differences in their levels of expression of the dopamine transporter (DAT; SLC6A3). These differences in DAT are strong candidates to contribute to individual differences in motor, mnemonic and reward functions. To identify "cis"-acting genetic mechanisms for these individual differences, we have sought variants in 5' aspects of the human DAT gene and identified the haplotypes that these variants define. 2. We report (i) significant relationships between 5' DAT haplotypes and human individual differences in ventral striatal DAT expression assessed in vivo using [(11)C] cocaine PET and (ii) apparent confirmation of these results in studies of DAT expression in postmortem striatum using [(3)H] carboxyflurotropane binding. 3. These observations support the idea that cis-acting variation in 5' aspects of the human DAT/SLC6A3 locus contributes to individual differences in levels of DAT expression in vivo. 5' DAT variation is thus a good candidate to contribute to individual differences in a number of human phenotypes. PMID- 16710761 TI - Return to work and adjustment latitude among employees on long-term sickness absence. AB - INTRODUCTION: The aim was to study whether return to work (RTW) after long-term sickness absence is affected by adjustment latitude i.e. opportunities to adjust one's work to one's state of health by e.g. choosing among work tasks and deciding about work pace and working hours. We also studied whether the effect of adjustment latitude differed between those returning full-time and those returning part-time. METHODS: Differences between men and women were also studied. A questionnaire was sent to 5,590 salaried employees who had been on sick leave for at least 90 days in 2000. The year after, 2001, they received a questionnaire which included questions about work status, working conditions, adjustment latitude and health. RESULTS: The questionnaire was returned from 3056 persons. Among women 32% were fully back to work, 34% were partly back and 34% were still on sick leave. Comparable figures for men were 33%, 32% and 36%. CONCLUSION: For both men and women the likelihood to RTW increased with increasing number of opportunities to adjust. Adjustment latitude increased returning to part-time as well as full-time work. The study indicates that work organisation is important for RTW. PMID- 16710762 TI - Prevalence of risk factors for adverse pregnancy outcomes during pregnancy and the preconception period--United States, 2002-2004. AB - OBJECTIVES: To assess the prevalence of risk factors for adverse pregnancy outcome during the preconception stage and during pregnancy, and to assess differences between women in preconception and pregnancy. METHODS: Data from the 2002 and 2004 Behavioral Risk Factor Surveillance System, United States, were used to estimate the prevalence of selected risk factors among women 18-44 in the preconception period (women who wanted a baby in the next 12 months, and were not using contraception, not sterile and not already pregnant) with women who reported that they were pregnant at the time of interview. RESULTS: Major health risks were reported by substantial proportions of women in the preconceptional period and were also reported by many pregnant women, although pregnant women tended to report lower levels of risk than preconception women. For example, 54.5% of preconception women reported one or more of 3 risk factors (frequent drinking, current smoking, and absence of an HIV test), compared with 32.0% of pregnant women (p < .05). The difference in the prevalence of these three risk factors between preconception and pregnancy was significant for women with health insurance (52.5% in preconception vs. 29.4% in pregnancy, p < .05), but not for women without insurance (63.4% vs. 52.7%, p > .05). CONCLUSIONS: Women appear to be responding to messages regarding behaviors that directly affect pregnancy such as smoking, alcohol consumption and taking folic acid, but many remain unaware of the benefits of available interventions to prevent HIV transmission and birth defects. Although it appears that some women reduce their risk for adverse pregnancy outcomes after learning of their pregnancy, the data suggest that a substantial proportion of women do not. Furthermore, if such change occurs it is often too late to affect outcomes, such as birth defects resulting from alcohol consumption during the periconception period. Preconception interventions are recommended to achieve a more significant reduction in risk and further improvement in perinatal outcomes. PMID- 16710763 TI - Preconception care in international settings. AB - OBJECTIVES: This literature review briefly describes international programs, policies, and activities related to preconception care and resulting pregnancy outcomes. METHODS: Electronic databases were searched and findings supplemented with secondary references cited in the original articles as well as textbook chapters, declarations, reports, and recommendations. RESULTS: Forty-two articles, book chapters, declarations, and other published materials were reviewed. Policies, programs, and recommendations related to preconceptional health promotion exist worldwide and comprise a readily identifiable component of historic and modern initiatives pertaining to women's health, reproductive freedom, and child survival. CONCLUSIONS: The integration of preconception care services within a larger maternal and child health continuum of care is well aligned with a prevention-based approach to enhancing global health. PMID- 16710759 TI - Pathophysiology and therapy of experimental stroke. AB - 1. Stroke is the neurological evidence of a critical reduction of cerebral blood flow in a circumscribed part of the brain, resulting from the sudden or gradually progressing obstruction of a large brain artery. Treatment of stroke requires the solid understanding of stroke pathophysiology and involves a broad range of hemodynamic and molecular interventions. This review summarizes research that has been carried out in many laboratories over a long period of time, but the main focus will be on own experimental research. 2. The first chapter deals with the hemodynamics of focal ischemia with particular emphasis on the collateral circulation of the brain, the regulation of blood flow and the microcirculation. In the second chapter the penumbra concept of ischemia is discussed, providing a detailed list of the physiological, biochemical and structural viability thresholds of ischemia and examples of how these thresholds can be applied for imaging the penumbra. The third chapter summarizes the pathophysiology of infarct progression, focusing on the role of peri-infarct depolarisation, the multitude of putative molecular injury pathways, brain edema and inflammation. Finally, the fourth chapter provides an overview of currently discussed therapeutic approaches, notably the effect of mechanical or thrombolytic reperfusion, arteriogenesis, pharmacological neuroprotection, ischemic preconditioning and regeneration. 3. The main emphasis of the review is placed on the balanced differentiation between hemodynamic and molecular factors contributing to the manifestation of ischemic injury in order to provide a rational basis for future therapeutic interventions. PMID- 16710764 TI - The history of preconception care: evolving guidelines and standards. AB - This article explores the history of the preconception movement in the United States and the current status of professional practice guidelines and standards. Professionals with varying backgrounds (nurses, nurse practitioners, family practice physicians, pediatricians, nurse midwives, obstetricians/gynecologists) are in a position to provide preconception health services; standards and guidelines for numerous professional organizations, therefore, are explored. The professional nursing organization with the most highly developed preconception health standards is the American Academy of Nurse Midwives (ACNM); for physicians, it is the American College of Obstetricians and Gynecologists (ACOG). These guidelines and standards are discussed in detail. PMID- 16710765 TI - Birth characteristics associated with early intervention referral, evaluation for eligibility, and program eligibility in the first year of life. AB - OBJECTIVES: The Individual with Disabilities Education Act mandates Early Intervention (EI) services for infants and toddlers with developmental delay. We assessed the percentage of infants at risk for developmental delay due to characteristics present at birth who were referred to Massachusetts EI within 1 year of birth, evaluated for eligibility, and eligible for services. In addition, we identified birth characteristics that independently predicted 0-1 year program referral, evaluation, and eligibility. METHODS: The Pregnancy to Early Life Longitudinal (PELL) data system linked birth certificate, hospital discharge, and EI program data of 219,037 infants born in Massachusetts, 1998-2000. Multivariate logistic regressions identified independent infant predictors of referral, evaluation, and eligibility. RESULTS: Of 219,037 births, 14,852 (6.8%) were referred to EI within 1 year. Birthweight<1200 g (OR=9.7, 95% CI 3.3-12.9) and birthweight 1200-1499 g (OR=7.4, 95% CI 5.8-9.5) strongly predicted referral. Referral was high (88%) among infants with two or more birth risks. Among referrals, 88% were evaluated for eligibility. The strongest predictor of evaluation was triplet birth (OR=4.3, 95% CI 1.6-11.8). Among infants evaluated, 85% were determined to be eligible. CONCLUSION: EI referral and evaluation are high among infants born at risk for delay in Massachusetts. Some characteristics not included in the state's high-risk definition (e.g. birthweight 1200-1499 g) were identified that predicted referral. Most referrals were eligible for services. Results demonstrate the value to states of using linked population and program data for program evaluation. PMID- 16710766 TI - Individualized performance feedback to increase prenatal domestic violence screening. AB - OBJECTIVES: Universal domestic violence (DV) screening once per trimester of pregnancy is recommended but rarely accomplished. Clinical leaders in this setting sought to improve adherence with this protocol. This prospective study used medical record audit and individualized performance feedback with peer comparison (IPF) to improve DV screening among first and second year obstetrics and gynecology (ob/gyn) residents. METHODS: The setting is a northeastern, urban, hospital-based, prenatal clinic serving low-income women. Most patients are Latina (75%); 11% are black and 9% are white. Few begin care in the first trimester (8.5%). We gave all residents DV training. Next we gave IPF-four reports at seven-week intervals. We reviewed medical record notes on patient visits corresponding to the first medical encounter and week 16 and week 28 of pregnancy. We used this data to compare screening immediately before IPF and following each IPF report. RESULTS: Screening increased steadily over time, from 60% of appropriate visits before IPF to 91% after the fourth report (Chi Square 28.4, p<.001). Adjusting for key factors, the odds of screening after the last IPF report were seven and a half times greater than the odds of screening before IPF (Odds Ratio: 7.6; 95% Confidence Interval: 3.0, 19.0). CONCLUSIONS: IPF was associated with increased DV screening among first and second year ob/gyn residents in this setting. Increased screening improved compliance with the clinic protocol and increased opportunities for patient disclosure, education, and treatment, critical public health objectives. PMID- 16710767 TI - Cloning and characterization of a novel human prefoldin and SPEC domain protein gene (PFD6L) from the fetal brain. PMID- 16710769 TI - Pheromone-induced priming of a defensive response in Western flower thrips. AB - The Western flower thrips Frankliniella occidentalis produces conspicuous anal droplets that function as a direct defense against various predators. These droplets also function in pheromonal communication in that they contain a mixture of decyl acetate and dodecyl acetate, which acts as an alarm. Exposure of thrips to synthetic pheromone is known to promote takeoff or refuge seeking, but the effect of the natural pheromone has not yet been studied. Here, we not only studied the response to natural pheromone, but also tested the new hypothesis that the alarm pheromone primes a defensive response in thrips. This test was carried out by measuring the reaction time to a simulated predator attack after exposure to synthetic or natural alarm pheromone (against a control with no pheromone at all). The reaction was quantified in terms of the time it takes a thrips larva to produce a droplet after attack. We found that thrips larvae produce droplets of alarm pheromone faster when cues associated with danger are present. There were no significant differences in reaction times of responses to synthetic pheromone, natural pheromone, or odors from a patch with a predator attacking a thrips larva. This implies that the synthetic pheromone mimics the natural pheromone, and that other cues emanating from the predator play a minor role. We conclude that the alarm pheromone increases the vigilance of the thrips, and this may promote its survival. PMID- 16710775 TI - Assortative mating for cigarette smoking and for alcohol consumption in female Australian twins and their spouses. AB - BACKGROUND: Non-random mating affects population variation for substance use and dependence. Developmentally, mate selection leading to positive spousal correlations for genetic similarity may result in increased risk for substance use and misuse in offspring. Mate selection varies by cohort and thus, assortative mating in one generation may produce marked changes in rates of substance use in the next. We aim to clarify the mechanisms contributing to spousal similarity for cigarette smoking and alcohol consumption. METHODS: Using data from female twins and their male spouses, we fit univariate and bivariate twin models to examine the contribution of primary assortative mating and reciprocal marital interaction to spousal resemblance for regular cigarette smoking and nicotine dependence, and for regular alcohol use and alcohol dependence. RESULTS: We found that assortative mating significantly influenced regular smoking, regular alcohol use, nicotine dependence and alcohol dependence. The bivariate models for cigarette smoking and alcohol consumption also highlighted the importance of primary assortative mating on all stages of cigarette smoking and alcohol consumption, with additional evidence for assortative mating across the two stages of alcohol consumption. CONCLUSIONS: Women who regularly used, and subsequently were dependent on cigarettes or alcohol were more likely to marry men with similar behaviors. After mate selection had occurred, one partner's cigarette or alcohol involvement did not significantly modify the other partner's involvement with these psychoactive substances. PMID- 16710776 TI - Genetic and environmental influences on age at sexual initiation in the Colorado Adoption Project. AB - Whereas the majority of research on adolescent sexual initiation has focused solely on environmental factors, the present study used behavioral genetic analyses to investigate the relative contributions of genetic and environmental influences. Structural equation models were fitted to data from adoptive and non adoptive sibling pairs (231 biologically related pairs and 169 unrelated pairs) from the Colorado Adoption Project. Information from censored individuals who had not yet experienced sexual initiation was maximized by adapting the twin survival analysis method of Pickles et al. (Behav Genet 24(5):457-468, 1994) to accommodate adoptive and non-adoptive siblings. Point estimates of variance components from an ACE model, including additive genetic (A), shared environmental (C), and non-shared environmental (E) influences were 28%, 24%, and 48%, respectively. Despite the lower point estimate for shared environmental effects than additive genetic effects, a CE model provided the best fit to the data. However, because adoptive siblings provide a direct estimate of shared environmental influences there is greater power to detect shared environmental effects in adoption designs. Evidence for genetic influences from our data were somewhat lower than those obtained in previous twin studies, possibly reflecting a return to more socially conservative sexual attitudes, changing sexual behaviors, or ambiguities in the wording of questions commonly used in research on adolescent sexuality. PMID- 16710777 TI - Identification of informative strains and provisional QTL mapping of amphetamine (AMPH)-induced locomotion in recombinant congenic strains (RCS) of mice. AB - Amphetamine (AMPH)-induced locomotor activity is a rodent behavioral trait that reflects mesolimbic dopaminergic activity. To identify potential quantitative trait loci (QTL) associated with this behavior, we used 34 recombinant congenic strains (RCSs) of mice derived from A/J (A strains) and C57BL/6J (B strains) and measured AMPH-induced total distance traveled (AMPH-TDIST). Two strains in the A panel (A52 and A63) showed significantly elevated AMPH-TDIST compared to the parental A/J strain and behaved similarly to C57BL/6J. Simple sequence length polymorphism (SSLP) markers on chromosomes 1, 2, 3, 5, 6, 8, 9, 10 and 20 were significantly associated with AMPH-TDIST in the A strains. Within the B panel, two strains (B81 and B74) had significantly higher and two strains (B69 and B75) had significantly lower AMPH-TDIST than C57BL/6J. Markers associated with AMPH TDIST in the B strains appeared on chromosomes 5, 17 and 20. Combining data from this approach and other genetic (mapping data in humans) and functional (cDNA expression) sources may help to identify suitable candidate genes relevant to human disorders where mesolimbic dopamine dysregulation has been postulated. PMID- 16710778 TI - In vivo characterization of brain morphometric and metabolic endophenotypes in three inbred strains of mice using magnetic resonance techniques. AB - C57BL6J, FVB/N and 129/SvJ mice are commonly used as background strains to engineer genetic models of brain pathologies and psychiatric disorders. Magnetic resonance imaging (MRI) and spectroscopy provide alternative approaches to neuroanatomy, histology and neurohistochemistry for investigating the correlation between genes and brain neuroanatomy and neurometabolism in vivo. We used these techniques to non-invasively characterize the cerebral morphologic and metabolic endophenotypes of inbred mouse strains commonly used in neurological and behavioral research. We observed a great variability in the volume of ventricles and of structures involved in cognitive function (cerebellum and hippocampus) among these strains. In addition, distinct metabolic profiles were evidenced with variable levels of N-acetylaspartate, a neuronal marker, and of choline, a compound found in membranes and myelin. Besides, significant differences in high energy phosphates and phospholipids were detected. Our findings demonstrate the great morphologic and metabolic heterogeneity among C57BL/ 6J, FVB/N and 129/SvJ mice. They emphasize the importance of selecting the appropriate genetic background for over-expressing or silencing a gene and provide some directions for modeling symptoms that characterize psychiatric disorders such as autism, schizophrenia and depression. PMID- 16710779 TI - Challenges in genetic studies of the etiology of substance use and substance use disorders: introduction to the special issue. AB - Misuse of psychoactive substances is associated with substantial costs to users and to society. A growing literature suggests individual differences in vulnerability to develop substance related problems are influenced to a large degree by genetic factors. We review the evidence from genetic epidemiologic and molecular genetic studies of problematic use of alcohol, tobacco, and other drugs, then discuss the challenges for the next generation of studies of genetic influences on substance use. These challenges are addressed in the remaining papers of this special issue. The papers cover a variety of approaches, substances, and non-human as well as human studies, but are united by their focus on going beyond heritability estimates to address the mechanisms and processes underlying the development of substance use and substance related problems, including measurement, precursors of substance abuse, stages of substance involvement, and specificity of genetic influences. PMID- 16710780 TI - Frequentist model-averaged estimators and tests for univariate twin models. AB - Parameter estimates from analyses of univariate twin data usually do not reflect the uncertainty due to the model selection phase of the data analysis. To address the effect of model selection uncertainty on parameter estimates, we introduce frequentist model-averaged estimators for univariate twin data analysis that use information-theoretic criteria to assign model weights. We conduct simulation studies to examine the performance of model-averaged estimators of additive genetic variance, and for tests for additive genetic variance based on model averaged estimators. In simulation studies with small or moderate sample sizes, model-averaged estimators of additive genetic variance typically have lower mean squared error than either (i) estimators from individual twin models, or (ii) estimators obtained from a decision procedure where the best-fitting model from likelihood-ratio testing is used to estimate additive genetic variance. For each sample size simulated, bootstrap tests based on model-averaged estimators have higher power to detect additive genetic variance than currently-used tests in most cases. PMID- 16710788 TI - The effects of attribution of responsibility and work history on perceptions of reasonable accommodations. AB - Under the Americans with Disabilities Act (ADA), employers must provide employees with disabilities reasonable accommodations that will enable them to perform job duties, as long as the accommodations do not financially burden the organization. Two studies were conducted to investigate whether disability origin and/or prior work history impermissibly influence the granting of reasonable accommodations under the ADA. In both studies, participants granted more accommodations for employees whose disability was caused by some external factor than for those whose disability was caused by the employee's own behavior. In Study Two, participants also granted more and costlier accommodations for an employee with an excellent work history than for an employee with an average work history. Implications of the use of extralegal factors in accommodation decisions are discussed. PMID- 16710790 TI - Early self-reported impairments in arm functioning of primary breast cancer patients predict late side effects of axillary lymph node dissection: results from a population-based cohort study. AB - OBJECTIVES: Improvements in the life expectancy of women with breast cancer raise important questions how to improve quality of life (QoL) for women sustaining complications and side effects of cancer treatment. The presented study examined the prevalence of arm morbidity in a cohort of primary breast cancer patients over time as a result of the extent of axillary lymph node dissection. Of particular interest is the question of using a recognized QoL assessment instrument at defined assessment points as an endpoint criteria of oncological treatment. METHODS: A prospective, population-based, longitudinal cohort study of patients with primary breast cancer was performed (n = 389). QoL data (EORTC QLQ C30 + BR23) and clinical data were assessed at designated time points. Primary endpoint of this analysis was patient reported arm morbidity assessed with the three-idem scale in the BR 23 (swelling, moving, pain). RESULTS: 20% of the patients evidenced considerable impairments in arm functioning. Arm morbidity was significantly related to the number of lymph nodes dissected (P < 0.002 entire cohort, P < 0.001 lymph node negatives) and was independent of age, stage of the disease, kind of breast surgery and radiation treatment. Early impairments in arm functioning (below 50 score values) assessed within 6 months after axillary surgery was a good predictor for late arm morbidity at 12 months RR 11.5 (CI 95% 4.7-28.4), 24 months RR 6.0 (CI95% 2.8-13.3) and 36 months RR 3.8 (CI 95% 1.8 7.9). CONCLUSIONS: Arm morbidity after axillary surgery is a severe and chronic condition affecting many breast cancer patients. The recognized QoL assessment instrument depict patients with severe impairments in arm functioning after axillary lymph node dissection and predict late arm morbidity. To increase patients' quality of life it thus may serve as a valid assessment tool for screening, allowing early referral for treatment and monitoring. PMID- 16710791 TI - Watering, fertilization, and slurry inoculation promote recovery of biological crust function in degraded soils. AB - Biological soil crusts are very sensitive to human-induced disturbances and are in a degraded state in many areas throughout their range. Given their importance in the functioning of arid and semiarid ecosystems, restoring these crusts may contribute to the recovery of ecosystem functionality in degraded areas. We conducted a factorial microcosm experiment to evaluate the effects of inoculation type (discrete fragments vs slurry), fertilization (control vs addition of composted sewage sludge), and watering frequency (two vs five times per week) on the cyanobacterial composition, nitrogen fixation, chlorophyll content, and net CO2 exchange rate of biological soil crusts inoculated on a semiarid degraded soil from SE Spain. Six months after the inoculation, the highest rates of nitrogen fixation and chlorophyll a content were found when the biological crusts were inoculated as slurry, composted sewage sludge was added, and the microcosms were watered five times per week. Net CO2 exchange rate increased when biological crusts were inoculated as slurry and the microcosms were watered five times per week. Denaturing gradient gel electrophoresis fingerprints and phylogenetic analyses indicated that most of the cyanobacterial species already present in the inoculated crust had the capability to spread and colonize the surface of the surrounding soil. These analyses showed that cyanobacterial communities were less diverse when the microcosms were watered five times per week, and that watering frequency (followed in importance by the addition of composted sewage sludge and inoculation type) was the treatment that most strongly influenced their composition. Our results suggest that the inoculation of biological soil crusts in the form of slurry combined with the addition of composted sewage sludge could be a suitable technique to accelerate the recovery of the composition and functioning of biological soil crusts in drylands. PMID- 16710792 TI - Zinc phosphate transformations by the Paxillus involutus/pine ectomycorrhizal association. AB - In this research, we investigate zinc phosphate transformations by Paxillus involutus/pine ectomycorrhizas using zinc-resistant and zinc-sensitive strains of the ectomycorrhizal fungus under high- and low-phosphorus conditions to further understand fungal roles in the transformation of toxic metal minerals in the mycorrhizosphere. Mesocosm experiments with ectomycorrhizas were performed under sterile conditions with zinc phosphate localized in cellophane bags: zinc and phosphorus mobilization and uptake by the ectomycorrhizal biomass were analyzed. In the presence of a phosphorus source, an ectomycorrhizal association with a zinc-resistant strain accumulated the least zinc compared to a zinc-sensitive ectomycorrhizal association and non-mycorrhizal plants. Under low-phosphorus conditions, mycorrhizal seedlings infected with the zinc-resistant strain increased the dissolution of zinc phosphate and zinc accumulation by the plant. Extended X-ray absorption fine structure analysis of both mycorrhizal and nonmycorrhizal roots showed octahedral coordination of zinc by oxygen-containing ligands such as carboxylates or phosphate. We conclude that zinc phosphate solubilization and zinc and phosphorus uptake by the association depend on ectomycorrhizal infection, strain of the mycobiont, and the phosphorus status of the matrix. PMID- 16710796 TI - Recognition and prevention of computed radiography image artifacts. AB - Initiated by complaints of image artifacts, a thorough visual and radiographic investigation of 197 Fuji, 35 Agfa, and 37 Kodak computed radiography (CR) cassettes with imaging plates (IPs) in clinical use at four radiology departments was performed. The investigation revealed that the physical deterioration of the cassettes and IPs was more extensive than previously believed. It appeared that many of the image artifacts were the direct result of premature wear of the cassettes and imaging plates. The results indicate that a quality control program for CR cassettes and IPs is essential and should include not only cleaning of the cassettes and imaging plates on a regular basis, but also visual and radiographic image inspection to limit the occurrence of image artifacts and to prolong the life cycle of the CR equipment. PMID- 16710797 TI - Developing a medical image content repository for e-learning. AB - The integration of medical informatics and e-learning systems could provide many advanced applications including training, knowledge management, telemedicine, etc. Currently, both the domains of e-learning and medical image have sophisticated specifications and standards. It is a great challenge to bring about integration. In this paper, we describe the development of a Web interface for searching and viewing medical images that are stored in standard medical image servers. With the creation of a Web solution, we have reduced the overheads of integration. We have packaged Digital Imaging and Communications in Medicine (DICOM) network services as a component that can be used via a Web server. The Web server constitutes a content repository for searching, editing, and storing Web-based medical image content. This is a simple method by which the use of Picture Archiving and Communication System (PACS) can be extended. We show that the content repository can easily interact and integrate with a learning system. With the integration, the user can easily generate and assign medical image content for e-learning. A Web solution might be the simplest way for system integration. The demonstration in this paper should be useful as a method of expanding the usage of medical information. The construction of a Web-based repository and integrated with a learning system may be also applicable to other domains. PMID- 16710798 TI - A multisite telemammography system for remote management of screening mammography: an assessment of technical, operational, and clinical issues. AB - OBJECTIVE: This paper describes a high-quality, multisite telemammography system to enable "almost real-time" remote patient management while the patient remains in the clinic. One goal is to reduce the number of women who would physically need to return to the clinic for additional imaging procedures (termed "recall") to supplement "routine" imaging of screening mammography. MATERIALS AND METHODS: Mammography films from current and prior (when available) examinations are digitized at three remote sites and transmitted along with other pertinent information across low-level communication systems to the central site. Images are automatically cropped, wavelet compressed, and encrypted prior to transmission to the central site. At the central site, radiologists review and rate examinations on a high-resolution workstation that displays the images, computer-assisted detection results, and the technologist's communication. Intersite communication is provided instantly via a messaging "chat" window. RESULTS: The technologists recommended additional procedures at 2.7 times the actual clinical recall rate for the same cases. Using the telemammography system during a series of "off-line" clinically simulated studies, radiologists recommended additional procedures at 1.3 times the actual clinical recall rate. Percent agreement and kappa between the study and actual clinical interpretations were 66.1% and 0.315, respectively. For every physical recall potentially avoided using the telemammography system, approximately one presumed "unnecessary" imaging procedure was recommended. CONCLUSION: Remote patient management can reduce the number of women recalled by as much as 50% without performing an unreasonable number of presumed "unnecessary" procedures. PMID- 16710801 TI - Detection of guinea pig macrophages by a new CD68 monoclonal antibody, PM-1K. AB - A new monoclonal antibody, PM-1K, was raised against 24-h cultured human peritoneal macrophages. In immunohistochemical assays, PM-1K recognized freshly isolated blood monocytes and most tissue macrophages as well as myeloid dendritic cells such as Langerhans cells and interdigitating cells. The molecular size of the antigen recognized by PM-1K was determined to be 110 kD by means of immunoaffinity purification. Because this affinity-purified antigen recognized by PM-1K was also recognized by anti-CD68 antibodies, it is believed to be one of the heterogeneous molecules of the CD68 antigen. Analysis showed interspecies reactivity of PM-1K with macrophages from guinea pigs, pigs, bovine species, and monkeys. Among these macrophages, those of the guinea pig reacted strongly with PM-1K. Patterns of PM-1K immunostaining in guinea pig tissues were similar to those found in human tissues. Studies with the immunoelectron microscope revealed reaction products of PM-1K in the cytoplasm, especially around endosomes. Since only a few antibodies are available to label guinea pig macrophages, PM-1K is considered to be one of the most suitable antibodies to examine macrophages in experimental guinea pig models. PMID- 16710803 TI - Four new species of the feather mite genus Vanginyssus Mironov, 2001 (Astigmata: Pteronyssidae) from the vangas (Passeriformes: Vangidae) in Madagascar. AB - Four new feather mite species of Vanginyssus Mironov, 2001 are described from vangas (Passeriformes: Vangidae), an endemic passerine family from Madagascar: Vanginyssus euryceros n. sp. from the helmet vanga Euryceros prevostii Lesson, V. madagascarinus n. sp. from the blue vanga Cyanolanius madagascarinus (Linnaeus), V. mystacornis n. sp. from the Crossley's babbler Mystacornis crossleyi (Grandidier) and V. orioliae n. sp. from the Bernier's vanga Oriolia bernieri Geoffroy Saint-Hilaire. A key to the five known species of Vanginyssus is provided. PMID- 16710806 TI - A simple homemade reaction station for use in parallel solution-phase synthesis. Optimization of a regioselective one-step deprotective o-formylation reaction mediated by the Vilsmeier-Haack reagent POCl3.DMF. AB - We report herein the fabrication of a simple and price-affordable portable reaction station for use in parallel solution-phase synthesis. This homemade device uses currently available laboratory components and equipment. Specifically designed to fit standard magnetic hotplates/stirrers, it can simultaneously hold up to 24 heated and magnetically stirred glass reactors of both 10 and 50 mL capacities. Glass reactors are connected by flexible 16-gauge metal needles to a central gas manifold equipped with an inlet/outlet for vacuum and inert gases. Reaction temperatures can be optimally varied from -78 ( composite function)C to 150 degrees C. Using a statistical screening DOE method, this parallel array reactor station has been successfully operated to optimize the one-step deprotective O-formylation of a sterically hindered bis-O-tert-butyldiphenylsilyl (O-TBDPS) aromatic diol. The latter transformation was mediated by the Vilsmeier Haack reagent POCl3.DMF using a range of Lewis acid and metal salt promoters, including their binary combinations. PMID- 16710807 TI - Toxicity of aliphatic ethers: a comparative study. AB - The CROMRsel procedure was used to model the toxicity of aliphatic ethers against mice. The best model obtained is based on three molecular descriptors and is a better model than other QSAR models from the literature. The only comparable model is one by Ren, based on four descriptors. PMID- 16710808 TI - A topological substructural molecular design to predict soil sorption coefficients for pesticides. AB - A TOPological Sub-structural MOlecular DEsign (TOPS-MODE) approach was used to predict the soil sorption coefficients for a set of pesticide compounds. The obtained model accounted for more than 85% of the data variance and demonstrated the importance of the dipole moment, the standard distance, the polarizability, and the hydrophobicity in describing the property under study. In addition, we compared this new model to a previous one using different descriptors such as WHIM and molecular connectivity indices. Finally, the TOPS-MODE was used to calculate the contribution of different fragments to the soil sorption coefficient of the compounds studied. The present approximation proved to be a good method for studying the soil sorption coefficient for pesticides, but it could also be extended to other series of chemicals. PMID- 16710809 TI - Modeling the bioconcentration factors and bioaccumulation factors of polychlorinated biphenyls with posetic quantitative super-structure/activity relationships (QSSAR). AB - During bioconcentration, chemical pollutants from water are absorbed by aquatic animals via the skin or a respiratory surface, while the entry routes of chemicals during bioaccumulation are both directly from the environment (skin or a respiratory surface) and indirectly from food. The bioconcentration factor (BCF) and the bioaccumulation factor (BAF) for a particular chemical compound are defined as the ratio of the concentration of a chemical inside an organism to the concentration in the surrounding environment. Because the experimental determination of BAF and BCF is time-consuming and expensive, it is efficacious to develop models to provide reliable activity predictions for a large number of chemical compounds. Polychlorinated biphenyls (PCBs) released from industrial activities are persistent pollutants of the environment that produce widespread contamination of water and soil. PCBs can bioaccumulate in the food chain, constituting a potential source of exposure for the general population. To predict the bioconcentration and bioaccumulation factors for PCBs we make use of the biphenyl substitution-reaction network for the sequential substitution of H atoms by Cl-atoms. Each PCB structure then occurs as a node of this reaction network, which is some sort of super-structure, turning out mathematically to be a partially ordered set (poset). Rather than dealing with the molecular structure via ordinary QSAR we use only this poset, making different quantitative super structure/activity relationships (QSSAR). Thence we developed cluster expansion and splinoid QSSARs for PCB bioconcentration and bioaccumulation factors. The predictive ability of the BAF and BCF models generated for 20 data sets (representing different conditions and fish species) was evaluated with the leave one-out cross-validation, which shows that the splinoid QSSAR (r between 0.903 and 0.935) are better than models computed with the cluster expansion (r between 0.745 and 0.887). The splinoid QSSAR models for BAF and BCF yield predictions for the missing PCBs in the investigated data sets. PMID- 16710810 TI - Structure-toxicity relationships of nitroaromatic compounds. AB - The toxicity data of 28 nitroaromatic compounds (nitrobenzenes and, for comparison, benzene and toluene) related to a 50% lethal dose concentration for rats (LD50) were used to develop quantitative structure-activity relationships (QSARs).A genetic algorithm and multiple regression analysis were applied to select the descriptors and to generate the correlation models. The obtained equations consist of one to three descriptors. A number of molecular descriptors was obtained from HF/6-31G(d) and DFT (B3LYP/6-311+G(d, p)) level calculations. The calculated molecular geometry and electronic properties were evaluated by comparison with the available experimental data (where applicable). All parameters obtained at the B3LYP/6-311+G(d, p) level and the topological descriptors derived from this geometry were found to be reliable, except for dipole moment, due to the large uncertainty of its estimation. Satisfactory relationships were observed for the one-parameter structure-toxicity models between topological (X5Av, Ms) and quantum-chemical (ELUMO) descriptors. For better predictability two- and three-parameter QSAR analyses were performed. These analyses resulted in much better equations with correlation coefficient values r = 0.872-0.924. These models have been obtained with a set of topological, fragment and quantum-chemical descriptors (Ms, PCR, PCD, BELe1, C 026 and ELUMO). The toxicity of nitroaromatic compounds appears to be governed by a number of factors, such as the number of nitrogroups, the electrotopological state, the presence of certain fragments and the electrophilicity/reactivity parameter (ELUMO). Nitrobenzenes exhibited electrophilic reactivity (as was shown by correlation of the toxicity with the energy of the lowest unoccupied orbital, ELUMO). The toxicity LD50 parameter for rats has been utilized for the first time for QSAR analysis of nitrobenzenes. The predictive ability of the models is determined by a cross-validation "leave-one-out" method. PMID- 16710811 TI - Successful computer guided planned synthesis of (4R)-thiazolidine carboxylic acid and its 2-substituted analogues as urease inhibitors. AB - By using internal combinatorial library we were able to identify (4R) thiazolidines carboxylic acid and its 2-substituted analogs as active inhibitors of urease. Molecular modeling and virtual screening were utilized to find out potential compounds. Computational techniques were employed at database of 90,000 ligands and selected the structure representing the low energy conformations, Grid and FlexX docking algorithms were used and the top binding ligands were synthesized and screened in wet-lab. PMID- 16710813 TI - Selective contribution of diabetes and other cardiovascular risk factors to cardiac autonomic dysfunction in the general population. AB - Both cardiac autonomic dysfunction adn cardiovascular risk factors are related to and excess risk of mortality. We sought to determine whether the major cardiovascular risk factors are associated with diminished heart rate variability (HRV), prolonged QTc interval, or increased QT dispersion (QTD). Male (n = 1030) and female (n = 957) subjects, aged 55-74 years, who participated in the population-based MONICA Augsburg survey 1989/90 were assessed for the presence of cardiovascular risk factors such as diabetes, hypertension, obesity, dyslipidemia, smoking, and low physical activity. Lowest quartiles for time domain indexes of HRV (SD of R-R intervals [SDNN], max-min difference), QTc > 440 ms, and QTD > 60 ms determined from 12-lead resting ECG were used as cutpoints. In men, after adjustment for age and alcohol consumption, significant independent determinants for the lowest quartiles of SDNN were diabetes, obesity, and smoking. Independent contributors to prolonged QTc were hypertension, obesity, smoking, and low physical activity, whereas for increased QTD it was only hypertension. In women, diabetes was the only contributor to low SDNN, and hypertension was the only determinant of prolonged QTc. In conclusion, diabetes is the primary determinant of reduced HRV in the general population, while hypertension is the primary contributor to prolonged QTc in both sexes. However, obesity and smoking contribute to autonomic dysfunction in men but not women. Thus, a selectivity and sex-related differences exist among the various cardiovascular risk factors as to their influence on autonomic dysfunction. PMID- 16710814 TI - [Non-haemorrhagic subependymal pseudocysts: ultrasonographic, histological and pathogenetic variability]. AB - PURPOSE: Differentiation of non-haemorrhagic subependymal pseudocysts from subependymal pseudocysts after cerebral haemorrhage in the preterm infant. MATERIALS AND METHODS: Selective ultrasonographic screening in 2200 neonates and retrospective analysis in 72 infants with subependymal pseudocysts, the full-term infants being analysed from birth, and the preterm infants after the second week of life, thus avoiding the gestational age at which cerebral haemorrhage occurs in the preterm infant. RESULTS: Three variants of pseudocysts were identified: the caudothalamic germinolysis is a leukomalacic and pseudocystic gliosis. Pseudocysts in the anterior choroid plexus of the lateral ventricle could be distinguished from caudothalamic germinolysis by their location, form and movement. Pseudocysts lateral of the frontal horns are the result of regression of germinal matrix remains. CONCLUSION: Differentiation of non-haemorrhagic from post-haemorrhagic germinolysis is necessary to clarify the aetiology and pathogenesis of non-haemorrhagic pseudocysts. Caudothalamic germinolysis possibly is the result of infection with stenotic intima proliferation following vasculitis. The results are thalamostriatal vasculopathy and germinal necrosis. Anterior plexus cysts might be the result of folding faults of the ependyma in the growth period of the choroid plexus. Pseudocysts lateral of the frontal horns should not be mistaken for ventricular ligaments. PMID- 16710815 TI - [Scope and limitations of diagnostics in breast disease--focussing on large core biopsy]. AB - Percutaneous high speed large core biopsy (LCB) using a 14-gauge-needle, supported by sonographic procedures, is a safe and reliable method for the diagnosis of benign and malignant breast lesions in daily practice. All malignant diagnoses (B5)--comprising 44 % (267/604) of cases--were confirmed by subsequent surgical biopsy. This indicates a positive predictive value of 100 %. Evidence for uncertain dignity in LCB (categories B3 and B4) was found in 3 % of our cases (20/604), requiring open breast surgery for further clarification. This is strongly supported by the fact that 22 % of B3 and 82 % of B4 cases were identified as carcinomas in subsequent surgery. In 9 % of our cases (53/604), histology was only questionably representative (category B1) without cancer in the follow-up. In the B2-category--which comprised nearly 44 % of all cases (n = 263)--the positive predictive value ranged at 2 %. For clinical investigation, mammography, breast ultrasound and LCB the sensitivity were at 79 %, 92 %, 92 % and 98 %, respectively. The specificity of clinical findings, mammography, breast ultrasound and LCB ranged at 88 %, 67 %, 79 % and 99 %, respectively. The combination of the 3 methods: clinical findings, mammography and breast ultrasound (leading to a "total diagnostic score" provided a sensitivity and specificity of 97 % and 52 %. LCB revealed an excellent specificity of 99 %. Therefore, large core biopsy takes up a definite and reliable place in procedures of breast diagnostics, thereby preventing a high number of unnecessary open surgical interventions. PMID- 16710816 TI - Clinical application of grey scale and colour Doppler sonography during abnormal third stage of labour. AB - AIM: The purpose of the study was to investigate whether colour Doppler sonography is helpful in the surveillance of abnormal third stage of labour. MATERIALS AND METHODS: 20 patients were enrolled in the prospective study. Inclusion criteria were third stage of labour > 15 min and/or clinical suspicion of retained placenta. By means of grey scale and colour Doppler sonography the length of distinct phases of third stage of labour and length of visualisation of blood flow between myometrium and placenta were measured. These data were compared with previously published normal values. RESULTS: The patients were allocated into four groups: 1. Patients with prolonged third stage of labour, but normal vaginal delivery of the placenta (Group 1, 8 cases). 2. Patients with clinically suspected retained placental parts (Group 2, 4 cases). 3. Patients with manual removal of the placenta without confirmation of placenta accreta (Group 3, 4 cases). 4. Patients with manual removal of the placenta with confirmation of placenta accreta (Group 4, 4 cases). A significant longer latent phase was responsible for the prolonged third stage of labour in Group 1 (p < 0.05). Blood flow between myometrium and placenta was significantly longer visible in Group 4 than in the normal cohort (p < 0.0001). CONCLUSION: Grey scale sonography can help to distinguish between uncomplicated and complicated prolonged third stage of labour. Colour Doppler sonography can detect persistent blood flow between myometrium and placenta during third stage of labour in cases of placenta accreta. In these instances, the patient may benefit from colour Doppler sonography-guided curettage. PMID- 16710817 TI - Real time 3D (4D) ultrasound-guided percutaneous biopsy of solid tumours. AB - PURPOSE: The aim of the current study was to investigate the feasibility of real time 3D (4D) ultrasound-guided biopsy. MATERIALS AND METHODS: Between 2003 and 2005, 53 patients with suspicious lymph nodes or solid tumours underwent 4D ultrasound-guided core needle biopsy using a 4D US-System (Voluson 730, GE/Kretztechnik, Austria). The results were documented prospectively and compared to a matched series of 53 conventional 2D ultrasound-guided core biopsies. RESULTS: 4D ultrasound-guided biopsy can be performed using multiplanar views or a combination of cross-sectional and rendered images. Assessment of additional 3D images during the biopsy procedure was helpful in 34 % of the cases. In the differentiation of malignant and benign lesions, 4D ultrasound guided biopsy yielded 35 true-positive results, 12 true-negative results and 5 false-negative results. The sensitivity and specificity for the diagnosis of malignancy was 87.5 % and 94.4 %, respectively (control group: 89.5 % and 94.4 %). There was one case of insufficient tissue sampling in the 4D targeting group and 2 cases in the control group. No complications were observed related to the biopsy procedure. CONCLUSIONS: 4D ultrasound-guided biopsy is feasible and can improve visualisation of biopsy procedures by multiplanar display or 3D rendering. Although the accuracy is comparable to that of conventional 2D biopsy, this technique may be helpful for biopsy of lesions with close relation to relevant anatomical structures. PMID- 16710818 TI - [Is there any role predictable for bone morphogenetic protein-7 in nephrology?]. AB - The existence of proteins with bone morphogenetic properties has been suggested for many years to explain the calcifying effect observed, with bone extracts, in ectopic tissues in vivo. Subsequently, at least thirty different proteins (identified as Bone Morphogenetic Proteins, BMPs) with this capacity have been discovered. The osteogenic activity of these substances is evident from the early phases of embryogenesis. However, besides bone, these proteins have been shown to affect other tissues too, where they affect cellular functions like proliferation, differentiation and apoptosis. They act through a specific receptor system partly shared with TGF-beta; in fact, they have been shown to inhibit TGF-beta actions. BMP-7 in particular is of vital importance in embryogenesis through its actions on bone, kidney and eyes, while in adulthood it is mainly expressed in the kidney, where specific receptors have been localized. BMP-7 has been successfully employed, for therapeutic purposes, in several animal models of nephropathies, where the most striking result is the constant reduction of fibrosis. Moreover, BMP-7 has been used successfully in experimental models of renal osteodystrophy and of vascular calcification. To date the only available human data are those obtained in orthopedy where BMP-7 has been successfully employed to induce the healing of pathologic fractures. Clinical data in nephropathic patients are therefore warranted to rule out the true value of this, very interesting, new molecule. PMID- 16710819 TI - [New insights in the treatment of ANCA-associated systemic vasculitis]. AB - The prognosis of untreated ANCA-associated systemic vasculitis (AASV) is poor with up to 90% of patients dying within 2 years. The combination of prednisone and cyclophosphamide is now established as the treatment of choice and leads to control of the disease in 80-90% of the patients. Such treatment has turned these acutely fatal diseases into chronic relapsing disorders with accumulating drug related morbidity in over 50% of the patients, including diabetes, bladder and lympho-proliferative malignancy and infertility. Treatment of AASV can be divided into three phases: therapy for induction of remission, for maintenance of remission and therapy for refractory and relapsing disease. In addition, the treatment must be tailored to the stage and severity of the disease and a new classification of AASV was introduced: localized vasculitis, early systemic vasculitis, generalized vasculitis, severe renal vasculitis and refractory vasculitis. Different randomized clinical trials have been performed with the aim of optimizing the existing therapeutic regimens: some of these have been concluded, others are still ongoing. Newer therapeutic approaches are currently being tested and have involved the use of mycophenolate mofetil, anti tumour necrosis factor (TNF) drugs (anti TNF antibody infliximab and humanized soluble TNF receptor etanercept), monoclonal anti- lymphocyte antibody (anti-CD20). These new alternative therapies could be used in patients with frequent relapses, in patients who fail to achieve remission with standard induction therapy and in those with severe side effects due to cumulative doses of cyclophosphamide. PMID- 16710820 TI - [Diagnostic and therapeutic criteria of arterial hypertension in childhood]. AB - Arterial hypertension (AH), either primary or secondary, is an important issue in childhood for its short- and long-term cardiovascular morbidity. Renal diseases are the most frequent causes of AH in children, but essential hypertension can also be detected early in life. It is important for blood pressure (BP) to be checked regularly (at least once every 5 years) in healthy children and adolescents and every year in those belonging to at-risk categories (family history of AH, low birth weight, obesity, etc). In children, AH is defined as BP recorded in three non-consecutive measurements with an appropriate device and cuff size > or = 95th centile for age, gender and height. Ambulatory BP monitoring is a valuable diagnostic tool and once AH is confirmed, a specific primary cause should always be ruled out (renovascular, cardiac, vascular, endocrine, pharmacologic, other). In case of border-line or significant AH (between 90th and 99th centile) a non-pharmacological treatment should be considered, whereas severe hypertension (>99th centile for height and age) will require pharmacological treatment (Diuretics, Angiotensin Converting Enzyme Inhibitors, Angiotensin Receptor Blockers, beta - and Calcium blockers). PMID- 16710821 TI - [Clinical relevance of study results]. AB - The critical appraisal process of available evidence includes the evaluation of clinical importance of the study findings. This should coincide with the minimum worthwhile effect expected by the investigators in the study design and planning. In hard outcome studies it can be quantified by the absolute risk difference between groups and its reciprocal, known as number needed to treat to avoid one adverse event, or benefit (NNTB). The number needed to treat to produce harmful consequences of treatment (NNTH) should also be taken into account. Finally these effect measures, like risks and incidence proportions, are usually rough estimates of the true effects, due to non-complete follow-up of the observations under study. Underlying assumptions and design issues are especially important to assess the clinical relevance of any results. PMID- 16710822 TI - [The Italian Society of Nephrology Guidelines (3rd Edition): principles and methods]. AB - Scientific Societies at both a local and international level are making big effort to prepare their clinical practice guidelines. The Italian Society of Nephrology has already published in two previous editions a series of guidelines relating to various aspects of management and diagnosis of different renal diseases. In this review we present the criteria of the 3(rd) edition of the Italian Society of Nephrology guidelines. This 3(rd) edition of guidelines will be based on the availability of scientific evidence in different areas of nephrology, dialysis and transplantation. Ten key intervention questions have been identified, based on the availability of systematic reviews of randomized trials or individual randomized address them. Systematic reviews and randomized trials are the optimal study design to address intervention questions. These have been summarized based upon rigid methodological criteria and strictly reflect the evidence basis. The different phases of development and publication of the 3(rd) edition of the Italian Society of Nephrology guidelines are presented. PMID- 16710824 TI - [Census of the Italian Nephrology and Dialysis Units. Comparison between Lombardy and Piedmont]. AB - The Italian Society of Nephrology promoted a national survey to obtain detailed information from all the Renal and/or Dialysis Units through an on-line questionnaire concerning structural, technological and human resources, as well as organisation characteristics and activities. The purpose of this initiative was to obtain regional reference benchmarks for each Nephrology Unit. In this paper we compare two northwestern Italian Regions: Lombardy and Piedmont. As far as epidemiology is concerned, the prevalence of dialysis patients is quite similar in the two Regions: for haemodialysis 616 pmp (patients per million population) in Lombardy and 595 in Piedmont, for peritoneal dialysis 104 pmp vs. 114 pmp, while the incidence of dialysis patients is 169 vs. 166 pmp. The gross mortality for dialysis patients is 12.4% vs. 13.7% and 0.9% vs. 2.0% in transplanted patients. The distribution of vascular access is also quite similar in the two Regions: prevalent arteriovenous fistula 83% vs. 74%, central venous catheter 11% vs. 18%, vascular grafts 7% vs. 8%. Structural resources: the hospital beds (49 pmp in the two Regions) and the dialysis places (161 vs. 166 pmp) do not differ between the two Regions. Personnel resources: physicians 37 pmp in Lombardy and 44 pmp in Piedmont, renal nurses 167 pmp vs. 186, respectively. Activity: hospital admission 1722 pmp vs. 1507 pmp, renal biopsies 131 pmp vs. 109 pmp. Although the two regions examined are numerically different, both have a high standard of quality, making Italy a model of nephrology organisation. This initiative to take a census of the Italian Nephrology and Dialysis Units provides an interesting tool to describe the present status of the operational structures, to identify precise benchmarking values, at both the regional and national level, and to act as a prelude for further rationalization and growth of the nephrology network in Italy. PMID- 16710823 TI - [Census 2004 of the Italian Renal and Dialysis Units. Veneto, Friuli Venezia Giulia and Trentino Alto Adige]. AB - The Italian Society of Nephrology (SIN) promoted a national survey of the Renal and Dialysis Units using an online questionnaire on some aspects of structural, technological and personnel resources, as well as organisation and activity. The major aim of this initiative was to obtain a reference benchmark on a national and regional basis. In this paper the data of the northestern regions of Italy (Veneto, Friuli-Venezia Giulia and Trentino-Alto Adige) are reported and compared with the recently published results of the northwestern regions (Piedmont, Liguria and Valle d'Aosta). From an epidemiologic point of view, the prevalence of dialysis patients was 534 pmp (per million population) in Veneto, 667 pmp in Friuli VG and 545 in Trentino AA, the prevalence of transplanted patients was 265, 294 and 404 pmp, respectively; the incidence of dialysis patients was 137, 182 and 130 pmp; gross mortality was 12.5, 14.3 and 16.5%; the distribution of vascular accesse in prevalently dialysis patients was: arteriovenous fistulas = 84.5, 70.4 and 80.9%, central venous catheters = 10.6, 20.0 and 10.2%, vascular graft = 4.9, 9.6 and 8.8%. Regarding structural resources, the distribution of hospital bed numbers was 38, 42 and 43 pmp; dialysis places were 137, 181 and 172 pmp. Human resources were given by renal physicians = 28.3, 38.2 and 23.6 pmp and renal nurses = 138, 200 and 172 pmp; each renal physician took care of 19, 17 and 23 dialysis patients and each renal nurse cared for 3.9, 3.3 and 3.2 dialysis patients. Activity data showed 1436, 1328 and 974 pmp hospital admissions, kidney biopsies were 106, 114 and 31 pmp. Overall, the Italian Northeast shows a significantly lower prevalence and incidence of end-stage renal disease patients than the Northwest; on the contrary, the incidence of patients with acute renal failure is significantly higher. In the Italian Northeast a significantly lower number of hospital beds devoted to renal patients is observed, while dialysis places are more frequent. In the Northeast fewer renal physicians are present than in the Northwest, whereas renal nurses are equivalent if related to the number of dialysis patients. Activity indexes, intended as amount of hospital admissions and renal biopsies standardised per population, are less significant in the Northeast. The results of the survey in Veneto, Friuli VG and Trentino AA show some discrepancies in the treatment of chronic kidney disease between the three regions and even more among different areas of Italy. Despite similar health care models, a relevant inequality in health care resources is evident. PMID- 16710825 TI - [Census 2004 of the Italian Renal and Dialysis Units. Emilia-Romagna, Toscana]. AB - The 2004 SIN census of the Italian nephrology and dialysis centres showed many interesting data about the epidemiology and the organization in the Regions of Emilia-Romagna (ER) and Tuscany (T). A) Epidemiology: incidence of dialysis patients 169 pmp (patients per million population) in ER, 147 ppm in T; prevalence of dialysis patients 639 pmp and 665 pmp, respectively; prevalence of transplanted patients 325 ppm in ER and 233 pmp in T; gross mortality of dialysis patients 16.3% and 13.4%, respectively; B) Type of vascular access in prevalently dialysis patients: arteriovenous fistula 83% and 78%; central venous catheter 13% and 12%; vascular graft 5% and 9%. C) Structural resources: nephrology beds 44 mp (per million population) and 50 mp; dialysis places 157 and 146 mp. D) Personnel resources : renal physicians 29 and 41 mp; renal nurses 171 and 202 mp ; each renal physician cares for 22 and 16 dialysis patients, and each renal nurse takes care of 3.7 and 3.3 dialysis patients. E) Activity: hospital admissions 1572, 1769 pmp; renal biopsies 115 and 166 pmp. PMID- 16710826 TI - [Historical Archives of Italian Nephrology. The form, evocative of function. A short historical review about the discovery of drugs from the most remote times to new trends in pharmacological sciences]. AB - The image, the imaginary and the imagined are more than ever the guide in pharmacological research, and predominate in the expectations of discovering, designing and producing a potential drug. The concept of ""form"" is now corroborated and implemented by the most recent and astounding acquisitions on the existing relationship in biology between form, structure and function. This article is a brief historical reconstruction of how the ""form"" has represented, from the most remote times, a fundamental guiding criterion in the choice of potential pharmacological remedies, and how in times of biotechnology and informatics the design and development of new drugs is based on the assumption that the form-function requisite is among the most promising trends in pharmacological sciences and drug discovery, in the nephrology field too. PMID- 16710827 TI - [From symptom to diagnosis: but not always!]. AB - The classic presentation of acute renal colic is the onset of severe flank pain associated to homolateral groin irradiation. Very often these patients present typical clinical diagnosis patterns. We present a case of nephrolithiasis with atypical characteristics. The exact diagnosis has been possible only after the release of the stone. PMID- 16710828 TI - [From the SIN Mailing List: the use of hemodialysis after radiocontrast media administration in patients with chronic renal failure]. AB - Over the last few years there has been a progressive increase in percutaneous endovascular procedures in patients with chronic renal disease, due to the high incidence of vascular disease, particularly coronary artery disease, in this population. The use of contrast medium may further worsen renal function in such patients, in some cases even accelerating the progression towards end-stage renal failure, and increase patients morbidity and mortality. In this review we discuss the role of dialysis in preventing contrast-induced nephropathy and indications to its use in patients already on dialysis treatment undergoing diagnostic or therapeutic procedures with contrast medium injection. PMID- 16710829 TI - [Resistant hypertension and paradox effect in patient with hemodialysis treatment]. AB - This is a case study of a 65-year-old female, on regular haemodialysis treatment, with resistant hypertension and paradoxical blood pressure (BP) elevation during dialysis. This phenomenon occurs in a small number of patients, since in most patients an acceptable BP is usually reached by adequate control of fluid and volume status with dialysis, sometimes associated with pharmacologic intervention. Since in our patient hypertension persisted despite apparent achievement of dry weight and maintenance of antihypertensive medications, we did some extensive investigations to disclose any secondary causes of hypertension (other than ESRD); we also evaluated whether the optimal dry weight was really achieved and maintained, and if the pharmacokinetics of the antihypertensive drugs was influenced by dialysis. We found no secondary cause of hypertension; by contrast, we detected the presence of a mild cardiac dilatation, and realized that some antihypertensive drugs, taken by our patient, were removed by dialysis. Since both these are known to precipitate the paradoxical BP rise during dialysis, we successfully modified once again our dialysis strategy and changed the antihypertensive therapy, adding a calcium antagonist to both losartan and low-dose minoxidil. PMID- 16710830 TI - Repression of CDC28 reduces the expression of the morphology-related transcription factors, Efg1p, Nrg1p, Rbf1p, Rim101p, Fkh2p and Tec1p and induces cell elongation in Candida albicans. AB - The ability of the human fungal pathogen Candida albicans to transit its cell shape is important for its pathogenicity. To obtain additional evidence that the cell cycle of C. albicans is associated with its morphology, we generated and characterized a conditional mutant of C. albicans CDC28, a cyclin-dependent kinase. In the constructed strain, the expression of CDC28 was regulated by the MET3 promoter and could be repressed in the presence of methionine and cysteine. Cdc28p-depleted cells demonstrated highly polarized growth and wider filaments than serum-induced hyphae. Hyphae-specific genes, HWP1, RBT4 and ECE1, were activated in the elongated filaments caused by the Cdc28p depletion. Furthermore, the protein expression levels of the transcription factors involved in morphological transition, Efg1p, Nrg1p, Rbf1p, Rim101p, Fkh2p and Tec1p, decreased under conditions that repress CDC28 expression. Taken together, these data indicate that repression of CDC28 affected the protein levels of the morphology-related transcription factors, the regulation of hyphae-specific genes and cell shape in C. albicans. PMID- 16710831 TI - Mak16p is required for the maturation of 25S and 5.8S rRNAs in the yeast Saccharomyces cerevisiae. AB - The nucleolar Mak16p protein of Saccharomyces cerevisiae has been implicated in 60S ribosome biogenesis. To learn more about the role of Mak16p in this process, ribosomal RNA processing was examined in a mak16-1 temperature-sensitive yeast strain. Steady-state levels of the 25S and 5.8S mature rRNA species dropped dramatically over a 4 h period in the mak16-1 yeast after a shift to the non permissive temperature, while 18S and 5S rRNA levels decreased only moderately. Ribosomal RNA processing (rRNA) analyses showed that the most prominent defect at the non-permissive temperature was a dramatic decrease in 27SB precursor RNA levels, with no significant increase in the levels of any precursor. These data indicate an essential role for Mak16p in the stability of the 27SB precursor rRNA. Association of Mak16p with the 66S preribosomal complex does not appear to be sufficient for its function, because the mutant Mak16-1p protein was detected in sucrose density gradient fractions corresponding to the 66S pre-RNP complex. PMID- 16710832 TI - Annotation of unknown yeast ORFs by correlation analysis of microarray data and extensive literature searches. AB - Changes in the expression of genes were used to elucidate the metabolic pathways and regulatory mechanisms that respond to environmental or genetic modifications. Results from previously published chemostat datasets were merged with novel data generated in the present study. ORFs displaying significant changes in expression that correlated with those of other ORFs were analysed using GO mapping tools and supplemented by literature information. The strategy developed was used to propose annotations for ORFs of unknown function. The following ORFs were assigned functions as a result of this study: YMR090w, YGL157w, YGR243w, YLR327c, YER121w, YFR017c, YGR067c, YKL187c, YGR236c (SPG1), YMR107w (SPG4), YMR206w, YER067w, YJL103c, YNL175C (NOP13) YJL200C, YDL070C (FMP16) and YGR173W. PMID- 16710833 TI - Prednisolone promotes remission and gastric mucosal regeneration in experimental autoimmune gastritis. AB - A cardinal feature of organ-specific autoimmunity is destructive pathology in the target organ. In human and experimental models of autoimmune gastritis, mononuclear cell infiltration and cellular destruction in the gastric mucosa are disease hallmarks. Strategies to cure autoimmune disease must not only establish immunological tolerance to autoantigen, but also rid the organ of pathogenic autoreactive cells. The present study has assessed the effect of prednisolone treatment in clearing the inflammatory infiltrate in experimental autoimmune gastritis and in preventing disease relapse in athymic compared with euthymic mice. Experimental autoimmune gastritis was induced by neonatal thymectomy or by transgenic expression of GM-CSF (PC-GMCSF mice). Groups of mice were treated with prednisolone (10 mg/kg per day) for 10 weeks or with prednisolone for 10 weeks followed by 10 weeks without prednisolone. Stomachs were examined for gross morphological changes, and by histology and immunohistochemistry for composition of inflammatory infiltrate and gastric mucosal integrity. Autoantibody to gastric H+/K+ ATPase was determined by ELISA. Prednisolone promoted remission of gastritis in both mouse models of experimental autoimmune gastritis, evident by reduction in stomach size, clearing of gastric inflammatory infiltrate, and regeneration of the gastric mucosa. Prednisolone withdrawal resulted in disease relapse in all PC-GMCSF mice, whereas approximately 40% of neonatal thymectomy mice retained normal stomach morphology and remained free of gastric pathology. It is concluded that prednisolone promotes remission and gastric mucosal regeneration in experimental autoimmune gastritis. Prolonged remission of autoimmune gastritis in some athymic mice suggests a role for the thymus in disease relapse. PMID- 16710834 TI - Near-infrared spectroscopic monitoring of biomass, glucose, ethanol and protein content in a high cell density baker's yeast fed-batch bioprocess. AB - The use of at-line NIRS to monitor a high cell density fed-batch baker's yeast bioprocess was investigated. Quantification of the key analytes (biomass, ethanol and glucose) and the product quality indicator (percentage protein content) was studied. Biomass was quantitatively modelled using whole matrix samples (as was percentage protein content). The dominance of the whole matrix spectrum by biomass, and its associated light scattering effects, were overcome by use of filtrate samples and adapted (semi-synthetic) filtrate samples, which allowed successful ethanol and glucose modelling, respectively. Calibrations were rigorously challenged via external validation with large sample sets relative to the calibration sample size, ensuring model robustness and potential practical utility. The standard errors of calibration for biomass, glucose, ethanol and total intracellular protein were (g/l) 1.79, 0.19, 0.79 and 0.91, respectively, comparable to those of the primary assays. The calibration strategies necessary to generate quantitative models for this range of analytes in such a complex high cell density bioprocess fluid are discussed. PMID- 16710835 TI - Towards treatment of viral pathogenesis. PMID- 16710836 TI - Human papillomavirus vaccines. AB - A wealth of epidemiological and molecular evidence has led to the conclusion that virtually all cases of cervical cancer and its precursor intra-epithelial lesions are a result of infection with one or other of a subset of genital human papillomaviruses (HPVs) suggesting that prevention of infection by prophylactic vaccination would be an effective anti-cancer strategy. The papillomaviruses cannot be grown in large amounts in culture in vitro, but the ability to generate HPV virus like particles (VLPs) by the synthesis and self-assembly in vitro of the major virus capsid protein L1 provides for a potentially effective sub unit vaccine. HPV L1 VLP vaccines are immunogenic and have a good safety profile. Published data from proof of principle trials and preliminary reports from large Phase III efficacy trials suggest strongly that they will protect against persistent HPV infection and cervical intra epithelial neoplasia. However, the duration of protection provided by these vaccines is not known, the antibody responses induced are probably HPV type specific and immunisation should occur pre-exposure to the virus. Second generation vaccines could include an early antigen for protection post-exposure and alternative delivery systems may be needed for the developing world. PMID- 16710837 TI - Adenovirus: from foe to friend. AB - Human adenoviruses (HAdVs) can cause mild respiratory, gastrointestinal, urogenital and ocular disease. Knowledge about HAdVs has been expanding for more than five decades putting them amongst the most-studied viruses. This continued interest stems, to a great extent, from the fact that these double-stranded DNA viruses have proven to be a versatile tool to probe the basic phenomena of eukaryotic cells. HAdV research has led to the discovery of, for instance, RNA splicing and greatly contributed to our knowledge of processes as fundamental as replication, transcription and translation. Moreover, the transformation of rodent cells by HAdVs has provided a system to unravel the molecular pathways that control cell proliferation. As a result, the genetic organisation of these agents is known in great detail allowing the straightforward manipulation of their genomes. In addition, the virus itself became renowned for its ability to produce large amounts of progeny and to efficiently infect mammalian cells regardless of their cell cycle status. These features contributed to the broad use of recombinant HAdVs as gene carriers particularly in in vivo settings where the vast majority of target cells are post-mitotic. The most advanced type of HAdV vectors can accommodate up to 37 kb of foreign DNA and are devoid of viral genes. With the aid of these high-capacity HAdV vectors large physiologically responsive transcriptional elements and/or genes can be efficiently introduced into target cells while minimising adaptive immune responses against the transduced cells. This article provides information on HAdV especially on the aspects pertinent to the design, production and performance of its recombinant forms. The development and characteristics of the main HAdV-based vector types are also briefly reviewed. PMID- 16710839 TI - Kyasanur forest disease: an epidemiological view in India. AB - Kyasanur forest disease (KFD) was first recognised as a febrile illness in the Shimoga district of Karnataka state of India. The causative agent, KFD virus (KFDV), is a highly pathogenic member in the family Flaviviridae, producing a haemorrhagic disease in infected human beings. KFD is a zoonotic disease and has so far been localised only in a southern part of India. The exact cause of its emergence in the mid 1950s is not known. A variant of KFDV, characterised serologically and genetically as Alkhurma haemorrhagic fever virus (AHFV), has been recently identified in Saudi Arabia. KFDV and AHFV share 89% sequence homology, suggesting common ancestral origin. Homology modelling of KFDV envelope (E) protein exhibited a structure similar to those of other flaviviruses, suggesting a common mechanism of virus-cell fusion. The possible mechanism of receptor-ligand interaction involved in infection by KFDV may resemble that of other flavivirses. Present understanding is that KFDV may be persisting silently in several regions of India and that antigenic and structural differences from other tick borne viruses may be related to the unique host specificity and pathogenicity of KFDV. From January 1999 through January 2005, an increasing number of KFD cases have been detected in Karnataka state of Indian subcontinent despite routine vaccination, suggesting insufficient efficacy of the current vaccine protocol. However, the exact cause of the increase of KFD cases needs further investigation. Considering the requirement of safer and more effective vaccines in general, there is clearly a need for developing an alternative vaccine as well as a rapid diagnostic system for KFD. The changing ecology of the prime focus of the KFD also warrants attention, as it may lead to establishment of the disease in newer localities, never reported before. PMID- 16710840 TI - Vaccinia virus proteolysis--a review. AB - It is well known that viruses, as obligate intracellular parasites, must use their hosts' metabolic machinery in order to replicate their genomes and form infectious progeny virions. What is less well known are the details of how viruses make sure that once all the necessary proteins are made, that they assume the correct configuration at the proper time in order to catalyse the efficient assembly of infectious virions. One of the methods employed by viruses to regulate this process is the proteolytic cleavage of viral proteins. Over the past several decades, studies in numerous laboratories have demonstrated that morphogenic proteolysis plays a major and essential role during the assembly and maturation of infectious poxvirus virions. In this review we describe the history of vaccinia virus proteolysis as a prototypic viral system. PMID- 16710841 TI - MMTV-like sequences in human breast cancer: a fluorescent PCR/laser microdissection approach. AB - The hypothesis that a retrovirus homologous to the mouse mammary tumour virus (MMTV) is involved in human breast cancer aetiology has fascinated scientists from many years, but it has never been convincingly demonstrated. Renewed interest in this hypothesis developed when an MMTV env gene-like sequence was found in 38% of human breast cancer tissues. Whereas some subsequent studies confirmed these findings, others did not. The main reasons for this discrepancy, among others, are the different sensitivities and technical details of current molecular approaches to the detection of these sequences. This study is an attempt to find sensitive and reproducible conditions capable of detecting MMTV env-like sequence in human samples. To this end, we first developed a fluorescence nested-PCR (FN-PCR) method that was able to detect very low copies of the viral genome, and then screened a panel of 45 frozen breast cancer samples obtained by laser microdissection. The MMTV env gene-like sequence was found in 15 (33%) of the human breast cancers analysed, whereas the same sequence was detectable neither in normal tissues nor in other types of tumour. Sequence analysis revealed 96% homology with the MMTV genome, but no other significant similarities with the human genome. The combined use of frozen material, microdissected cell populations and FN-PCR provides a novel, sensitive, robust, non-radioactive and fast methodology for the molecular detection of human-MTV. This approach might be successfully used in large molecular studies that aim to investigate the hypothesis of a retroviral aetiology of human breast cancer. PMID- 16710842 TI - The antibody 2B4 directed against the Epstein-Barr virus (EBV)-encoded nuclear antigen 1 (EBNA1) detects MAGE-4: implications for studies on the EBV association of human cancers. AB - We have previously developed two monoclonal antibodies against the Epstein-Barr Virus (EBV) nuclear antigen 1 (EBNA1), designated 1H4 and 2B4. Both detect EBNA1 by in situ staining in established EBV-positive tumours, e.g. Hodgkin's lymphoma and nasopharyngeal carcinoma. An association of EBV with other tumours, notably breast carcinomas, has been reported but remains controversial. Using the antibody 2B4, a nuclear protein has been detected in breast carcinomas that were EBV-negative by other methods, suggesting cross-reactivity with a cellular protein. Furthermore, an association of EBV with various other carcinomas has been reported on the basis of 2B4 immunohistochemistry. Here we show that 2B4 also binds to MAGE-4, a cancer testis antigen expressed in a variety of tumour cells, including breast carcinoma, seminoma and EBV-negative cases of Hodgkin's lymphoma. We conclude that the 2B4 antibody is not suitable for the detection of EBV infection but that additional techniques, particularly in situ hybridization for the detection of the EBV-encoded RNAs (EBERs), should be employed to confirm the presence of EBV. Our results add to the evidence indicating that breast cancer is not an EBV-associated disease. PMID- 16710843 TI - Transcriptional response to nitrosative stress in Saccharomyces cerevisiae. AB - Nitric oxide and NO-derived species (RNS) are defence molecules with broad antimicrobial activity. Microorganisms have developed strategies to sense RNS and counteract their damaging effects. We used Saccharomyces cerevisiae, harbouring a deletion of YHB1 that encodes the main NO scavenger enzyme, to study consequences of RNS exposure on whole-genome transcriptional response. The expression of > 700 genes was altered on RNS treatment. No major role for ROS-scavenging enzymes was found, and the respiratory chain, the main site of ROS production, had only minor involvement in the RNS-induced stress. The changes were generally transient and also found after treatment with the respiratory inhibitor myxothiazol. However, 117 genes showed a persistent response that was not observed after myxothiazol treatment. Of these, genes of the glutathione and DNA repair systems, iron homeostasis and transport were found to be upregulated. Severe repression of genes of respiratory chain enzymes was observed. Many of these genes are known to be regulated by the transcription factor Hap1p, suggesting that RNS might interfere with Hap1p activity. We showed also that Msn2/4p and Yap1p, key regulators of the response to general stress and oxidative stress, respectively, played a role in mediating the RNS-induced response. PMID- 16710844 TI - Jaw muscle functional anatomy in northern grasshopper mouse, Onychomys leucogaster, a carnivorous murid. AB - The jaw muscle anatomy of the northern grasshopper mouse, Onychomys leucogaster, was observed and the mechanical basis of the insectivorous/carnivorous adaptations were examined. Compared with Peromyscus maniculatus, a granivorous relative of Onychomys, there is a reduction of some aponeuroses within the masseter deep layer. This characteristic indicates that shearing meat or crushing arthropod exoskeletons requires less occlusal pressure than does grinding plant material. In Onychomys both the anterior and posterior portions of the masseter deep layer are more anterodorsally inclined, so that the line of action of the masseter lies further from the jaw joint than in Peromyscus. A strong incisal bite for killing vertebrates such as other rodents can be produced by a jaw mechanism with the high lever advantage of this muscle, which compensates for the decline in muscle mass. Our quantitative analysis suggests that the disappearance of an aponeurosis along the zygomatic plate in Onychomys decreases the stretch of the corresponding muscle, i.e., the anterior fibers of the masseter deep layer, accompanying jaw opening, and increases the maximum gape necessary for hunting large prey. PMID- 16710845 TI - Description of a cranial endocast from a fossil platypus, Obdurodon dicksoni (Monotremata, Ornithorhynchidae), and the relevance of endocranial characters to monotreme monophyly. AB - A digital cranial endocast of the Miocene platypus Obdurodon dicksoni was extracted from high-resolution X-ray computed tomography scans. This endocast represents the oldest from an unequivocal member of either extant monotreme lineage and is therefore important for inferring character support for Monotremata, a clade that is not well diagnosed. We describe the Obdurodon endocast with reference to endocasts extracted from skulls of the three species of extant monotremes, particularly Ornithorhynchus anatinus, the duckbill platypus. We consulted published descriptions and illustrations of whole and sectioned brains of monotremes to determine which external features of the nervous system are represented on the endocasts. Similar to Ornithorhynchus, well developed parafloccular casts and reduced olfactory bulb casts are present in the Obdurodon endocast. Reduction of the olfactory bulbs in comparison with tachyglossids and therian mammals is a potential apomorphy for Ornithorhynchidae. The trigeminal nuclei, ganglia, and nerves (i.e., trigeminal complex) are enlarged in Obdurodon, as evidenced by their casts on the endocast, as is the case in the extant platypus. The visibility of enlarged trigeminal nucleus casts on the endocasts of Obdurodon and Ornithorhynchus is a possible synapomorphy of Ornithorhynchidae. Electroreception and enlargement of the trigeminal complex are possible synapomorphies for Monotremata. PMID- 16710846 TI - Nitric oxide donor molsidomine attenuates psychotomimetic effects of the NMDA receptor antagonist MK-801. AB - There is experimental evidence indicating that the non-competitive NMDA receptor antagonist MK-801 impairs cognition and produces a series of schizophrenia-like symptoms in rodents (hypermotility, stereotypies, and ataxia). The present study was designed to investigate the efficacy of the nitric oxide (NO) donor molsidomine in counteracting these MK-801-induced behavioral effects in the rat. In a first study, post-training administration of molsidomine (at 4 but not 2 mg/kg) successfully antagonized MK-801-induced performance deficits in a recognition memory test. In a subsequent study, molsidomine (2 and 4 mg/kg) was shown to be unable to reverse MK-801-induced hypermotility but attenuated stereotypies (continuous movement whole cage, body sway, and head weaving) produced by MK-801. Moreover, at 4 mg/kg this NO donor counteracted MK-801 induced ataxia. Our findings indicate that molsidomine attenuates behavioral effects related to the hypofunction of the NMDA receptor suggesting that NO might be involved in the psychotomimetic effects of non-competitive NMDA receptor antagonists. PMID- 16710847 TI - Rewiring enervated: thinking LARGEr than myodystrophy. AB - LARGE is a glycosyltransferase known to glycosylate alpha-dystroglycan, a component of the dystrophin-associated glycoprotein complex. Spontaneous deletions in the Large gene (Large(myd) and Large(vls)) result in muscular dystrophy accompanied by heart, brain, and eye defects. Another Large mouse mutant, enervated (Large(enr)), is the result of a transgene integration event that disrupts Large gene expression. In addition to myodystrophy, enr mice have been shown to display peripheral nerve abnormalities, including altered axonal sorting resulting from Schwann cell defects, poor regeneration after nerve injury, and abnormal neuromuscular junctions. These data have provided new insight into our understanding of the function of LARGE and have suggested the possibility of involvement of substrates in addition to alpha-dystroglycan in the generation of the LARGE phenotype. The Large mutants are excellent models for addressing the importance of glycosylation in neuromuscular disease. PMID- 16710849 TI - Substitution and protonation effects on spin-spin coupling constants in prototypical aromatic rings: C6H6, C5H5N and C5H5P. AB - Ab initio equation-of-motion coupled cluster calculations have been carried out to evaluate one-, two-, and three-bond 13C-13C, 15N-13C, 31P-13C coupling constants in benzene, pyridine, pyridinium, phosphinine, and phosphininium. The introduction of N or P heteroatoms into the aromatic ring not only changes the magnitudes of the corresponding X-C coupling constants (J, for X = C, N, or P) but also the signs and magnitudes of corresponding reduced coupling constants (K). Protonation of the heteroatoms also produces dramatic changes in coupling constants and, by removing the lone pair of electrons from the sigma-electron framework, leads to the same signs for corresponding reduced coupling constants for benzene, pyridinium, and phosphininium. C-C coupling constants are rather insensitive to the presence of the heteroatoms and protonation. All terms that contribute to the total coupling constant (except for the diamagnetic spin-orbit (DSO) term) must be computed if good agreement with experimental data is to be obtained. PMID- 16710848 TI - Chronic systemic D-galactose exposure induces memory loss, neurodegeneration, and oxidative damage in mice: protective effects of R-alpha-lipoic acid. AB - Chronic systemic exposure of mice, rats, and Drosophila to D-galactose causes the acceleration of senescence and has been used as an aging model. The underlying mechanism is yet unclear. To investigate the mechanisms of neurodegeneration in this model, we studied cognitive function, hippocampal neuronal apoptosis and neurogenesis, and peripheral oxidative stress biomarkers, and also the protective effects of the antioxidant R-alpha-lipoic acid. Chronic systemic exposure of D galactose (100 mg/kg, s.c., 7 weeks) to mice induced a spatial memory deficit, an increase in cell karyopyknosis, apoptosis and caspase-3 protein levels in hippocampal neurons, a decrease in the number of new neurons in the subgranular zone in the dentate gyrus, a reduction of migration of neural progenitor cells, and an increase in death of newly formed neurons in granular cell layer. The D galactose exposure also induced an increase in peripheral oxidative stress, including an increase in malondialdehyde, a decrease in total anti-oxidative capabilities (T-AOC), total superoxide dismutase (T-SOD), and glutathione peroxidase (GSH-Px) activities. A concomitant treatment with lipoic acid ameliorated cognitive dysfunction and neurodegeneration in the hippocampus, and also reduced peripheral oxidative damage by decreasing malondialdehyde and increasing T-AOC and T-SOD, without an effect on GSH-Px. These findings suggest that chronic D-galactose exposure induces neurodegeneration by enhancing caspase mediated apoptosis and inhibiting neurogenesis and neuron migration, as well as increasing oxidative damage. In addition, D-galactose-induced toxicity in mice is a useful model for studying the mechanisms of neurodegeneration and neuroprotective drugs and agents. PMID- 16710850 TI - Epidermal growth factor receptor expression regulates proliferation in the postnatal rat retina. AB - Epidermal growth factor (EGF) is known to promote proliferation of both retinal progenitors and Muller glia in vitro, but several questions remain concerning an in vivo role for this factor. In this study, we investigated whether the EGF receptor (EGFR) is necessary for the maintenance of normal levels of progenitor and Muller glial proliferation in vivo. Here, we show that (1) mice with homozygous deletion of the Egfr gene have reduced proliferation in late stages of retinal histogenesis, (2) EGF is mitogenic for Muller glia in vivo during the first two postnatal weeks in the rodent retina, (3) the effectiveness of EGF as a Muller glial mitogen declines in parallel with the decline in EGFR expression as the retina matures, and (4) following damage to the retina from continuous light exposure, EGFR expression is up-regulated in Muller glia to levels close to those in the neonatal retina, resulting in a renewed mitotic response to EGF. Together with previous results from other studies, these data indicate that the downregulation of a growth factor receptor is one mechanism by which glial cells maintain mitotic quiescence in the mature nervous system. PMID- 16710851 TI - Effects of cold storage on the function and morphology of isolated urinary bladder in rat. AB - AIMS: We investigated the effects of 24- and 48-hr storage at 4 degrees C in Krebs solution on the function and morphology of isolated, rat urinary bladders. METHODS: Strips of bladder were obtained from eight male Sprague-Dawley rats. Six strips were harvested from each bladder and randomized to storage for 24 or 48 hr at 4 degrees C in Krebs solution or examination immediately after harvest. Contractile responses of the strips to potassium chloride (KCl), electric field stimulation (EFS), adenosine 5'-triphosphate (ATP) and carbamylcholine (CCh) were assessed. Histological examination of the bladder strips was performed. The pO(2), pCO(2), and pH of the solution in each storage container were measured at each storage time point. RESULTS: Cold storage induced a significant decrease in the amplitude of contraction in response to KCl and EFS after 24 or 48 hr of storage compared with control. The response of the bladder strips to ATP and CCh was significantly reduced after 48-hr storage compared with control, but not 24 hr storage. The pO(2) and pCO(2) decreased after cold storage. The pH increased after 24 hr of storage and remained stable between 24 and 48 hr of storage. Histological evaluation of the strips showed tissue swelling after 24 and 48 hr of storage. CONCLUSIONS: These results suggest that the morphology and function of bladder strips stored for 24 to 48 hr at 4 degrees C in Krebs solution undergo significant changes. Further studies are needed to assess the allowable time for storage of bladder tissue. PMID- 16710853 TI - A randomized, double-blind comparison of olanzapine/fluoxetine combination, olanzapine, fluoxetine, and venlafaxine in treatment-resistant depression. AB - Based on preliminary evidence of its usefulness in treatment-resistant depression (TRD), an olanzapine/fluoxetine combination (OFC) was examined in comparison with olanzapine, fluoxetine, and venlafaxine in a TRD population. In this 12-week double-blind study, 483 subjects with unipolar, nonpsychotic TRD, with historic failure on a selective serotonin reuptake inhibitor (SSRI) and prospective failure on open-label venlafaxine, were randomized to an OFC or to an olanzapine, fluoxetine, or venlafaxine monotherapy group. Venlafaxine was continued randomly in the double-blind acute phase to explore the benefits of continuation versus switching therapy. The Montgomery-Asberg Depression Rating Scale (MADRS) total change score at end point was the primary outcome measure. The OFC group had significantly greater improvement in depressive symptoms by week 1 of treatment (MADRS mean change =-7.2, baseline =29.6), in comparison to olanzapine (-4.8, P=.03), fluoxetine (-4.7, P=.03), or venlafaxine (-3.7, P=.002) groups and maintained its statistical separation from all three monotherapy groups through week 6. At end point, the OFC group was significantly different only from the olanzapine group (-14.1 vs. -7.7, P<.001). Analysis of a subgroup of subjects who had an SSRI failure in their current depressive episode (n=334) revealed statistical separation from both olanzapine and fluoxetine (but not venlafaxine) at end point: OFC (-14.6) versus olanzapine (-9.4, P<.001) versus fluoxetine ( 10.7, P=.006) versus venlafaxine (-14.7, P=.98). The OFC had a safety profile comparable to its component monotherapies (i.e., olanzapine and fluoxetine), showed a rapid onset of antidepressant effect, and was effective in this TRD sample. At the study end point, OFC, fluoxetine, venlafaxine, and low-dose OFC all appeared to be similarly effective. PMID- 16710854 TI - Quantitative survival model for short-term survival after adult-to-adult living donor liver transplantation. AB - Adult-to-adult living donor liver transplantation (ALDLT) has been accepted as an important option for end-stage liver disease, but information regarding the risk factors remains fragmentary. We aimed to establish a predictive model for 90-day survival. In the first step, a total of 286 cases who had received primary ALDLT using a right lobe graft between 1998 and 2004 were randomly divided into 2 cohorts at a ratio of 2:1 (191 vs. 95 recipients). The larger cohort of patients was used to develop a model. The outcome was defined as 90-day survival, and a total of 39 preoperative and operative variables, including the period of surgery (1998-2001 vs. 2002-2004), were included using Cox's proportional hazard regression model. Two mismatches of human leukocyte antigen (HLA) type DR (hazard ratio [HR] = 4.45; confidence interval [CI] = 1.96-10.1), log(e)[blood loss volume] (HR = 2.43; CI = 1.64-3.60), period of surgery (1998-2001 vs. 2002-2004) (HR = 2.41; CI = 1.04-5.57), and log(e)[serum C-reactive protein or CRP] (HR = 1.64; CI = 1.13-2.38) were found to be independent risk factors. In the second step, we tried to establish a realistic survival model. In this step, we created 2 models, 1 that used all 4 variables (model 1) and 1 (model 2) in which blood loss volume was replaced with the past history of upper abdominal surgery and Model for End-Stage Liver Disease (MELD) score (> or =25), both of which showed associations with blood loss volume. These models were applied to the smaller cohort of 95 patients. Receiver operating characteristic analyses demonstrated that both models showed similar significant c-statistics (0.63 and 0.62, respectively). In conclusion, model 2 can provide a rough estimation of the 90 day survival after ALDLT. PMID- 16710855 TI - Histological recurrent hepatitis C after liver transplantation: Outcome and role of retransplantation. AB - Impact of hepatitis C virus (HCV) recurrence on long-term outcome after orthotopic liver transplantation (OLT) is highly variable, and the role of retransplantation is still debated. From 1996 to 2003, 131 OLT with histologically proven HCV recurrence and 6 months of follow-up were retrospectively reviewed. One and 5-yr overall survivals were 90.7 and 81.3%, respectively. The mean time of HCV recurrence was 10.1 +/- 6.2 months in patients whose donor's age was less than 70 yr old, and 6.6 +/- 4.7 in patients whose donor's age was more than 70 (P < 0.01). The mean time between OLT and HCV recurrence was 10.7 +/- 8.2 months among patients still alive, and 5 +/- 4.2 among the 20 who died (P = 0.02). In 16 (12.2%) patients, retransplantation was required for severe HCV recurrence; 5 are still alive and 11 (68.7%) died. The mean survival time was 16.2 +/- 6 months if re-OLT was performed within 12 months from first OLT, and it was 45.9 +/- 10 months if re-OLT was performed later (P < 0.01). In conclusion, donors older than 70 yr are at high risk of early HCV recurrence; expectancy of life is significantly reduced in case of histologically proven recurrence within 6 months. Outcome is quite dismal in patients with early HCV recurrence requiring retransplantation within 1 yr of first OLT. PMID- 16710856 TI - Isolated liver transplantation in infants with end-stage liver disease due to short bowel syndrome. AB - Infants with short bowel syndrome (SBS) and associated liver failure are often referred for combined liver/intestinal transplantation. We speculated that in some young children, nutritional autonomy would be possible with restoration of normal liver function. Features we believed to predict nutritional autonomy include history of at least 50% enteral tolerance, age less than 2 yr, and no underlying intestinal disease. This report documents our experience with liver transplantation alone in children with liver failure associated with SBS. Twenty three children with SBS and end-stage liver disease, considered to have good prognostic features for eventual full enteral adaptation, underwent isolated liver transplantation. Median age was 11 months (range, 6.5 to 48 months). Median pretransplant weight was 7.4 kg (range, 5.2 to 15 kg). All had growth retardation and advanced liver disease. Bowel length ranged from 25 to 100 cm. Twenty-three children underwent 28 isolated liver transplants. There were 14 whole livers and 14 partial grafts (five living donors). Seventeen patients are alive at a median follow-up of 57 months (range, 6 to 121 months). Actuarial patient and graft survival rates at 1 yr are 82% and 75% and at 5 yr are 72% and 60%, respectively. Four deaths resulted from sepsis, all within 4 months of transplantation, and 1 death resulted from progressive liver failure. Two allografts developed chronic rejection; both children were successfully retransplanted with isolated livers. Of 17 surviving patients, three require supplemental intravenous support; the remaining 14 have achieved enteral autonomy, at a median of 3 months (range, 1 to 72 months) after transplantation. Linear growth is maintained and, in many, catch up growth is evident. Median change in z score for height is 0.57 (range, -4.47 to 2.68), and median change in z score for weight is 0.42 (range, -1.65 to 3.05). In conclusion, Isolated liver transplantation in children with liver failure as a result of SBS, who have favorable prognostic features for full enteral adaptation, is feasible with satisfactory long-term survival. PMID- 16710857 TI - Three-dimensional reconstruction of the nose epidermal cells in the microbial feeding nematode, Acrobeles complexus (Nematoda: Rhabditida). AB - The epidermis of the anterior end (nose) plays an important role in the evolution, development, and functional feeding morphology in nematodes, but information on this complex organ system is limited. Here, we produce a 3D model of 13 of the cells making up this organ system reconstructed from serial transmission electron micrographs of the microbial feeding nematode, Acrobeles complexus. Nose epidermal cells were found to be broadly similar to those of the distantly related model organism Caenorhabditis elegans in the number and arrangement of nuclei in these largely syncytial cells; this similarity demonstrates striking evolutionary conservation that allows for robust statements of homology between the taxa. Examining details of cell shape, however, revealed surprisingly complex subcellular specialization, which differed markedly from C. elegans in the number and arrangement of cell processes. Anterior toroid processes of the anterior arcade, posterior arcade, and HypB syncytia form a nested complex at the base of the labial probolae. Anterior toroid processes of HypC and the inner labial socket cells are associated with the base of the cephalic probolae and radial ridge processes. Extracellular filaments (tendon organs) and radiating cytoskeletal filaments of the posterior arcade syncytium form a connection between the body wall muscle cells and the pharynx. An epidermal cell with no known homolog in other nematodes is identified. Findings provide a basis to propose hypotheses related to the development and evolutionary origin of specialized feeding appendages (probolae) in the Cephalobinae (including Acrobeles), and hypotheses of homology are revised for epidermal cells in the nose of the closely related and primarily plant parasitic group, Tylenchida. PMID- 16710858 TI - Impact of preoperative steroids administration on ischemia-reperfusion injury and systemic responses in liver surgery: a prospective randomized study. AB - Hepatic injury secondary to warm ischemia-reperfusion (I/R) injury and alterations in haemostatic parameters are often unavoidable events after major hepatic resection. The release of inflammatory mediator is believed to play a significant role in the genesis of these events. It has been suggested that preoperative steroid administration may reduce I/R injury and improve several aspects of the surgical stress response. The aim of this prospective randomized study was to investigate the clinical benefits on I/R injury and systemic responses of preoperatively administered corticosteroids. Seventy-six patients undergoing liver resection were randomized either to a steroid group or to a control group. Patients in the steroid group received preoperatively 500 mg of methylprednisolone. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin, coagulation parameters, and inflammatory mediators, interleukin 6 and tumor necrosis factor alpha were compared between the 2 groups. Length of stay, and type and number of complications were recorded as well. Postoperative serum levels of ALT, AST, total bilirubin, and inflammatory cytokines were significantly lower in the steroid than in the control group at postoperative days 1 and 2. Changes in hemostatic parameters were also significantly attenuated in the steroid group. In conclusion, the incidence of postoperative complications in the steroid group tended to be significantly lower than the control group. It is of clinical interest that preoperative steroids administration before major surgery may reduce I/R injury, maintain coagulant/anticoagulant homeostasis, and reduce postoperative complications by modulating the inflammatory response. PMID- 16710859 TI - Therapeutic potential of sulfamides as enzyme inhibitors. AB - Sulfamide, a quite simple molecule incorporating the sulfonamide functionality, widely used by medicinal chemists for the design of a host of biologically active derivatives with pharmacological applications, may give rise to at least five types of derivatives, by substituting one to four hydrogen atoms present in it, which show specific biological activities. Recently, some of these compounds started to be exploited for the design of many types of therapeutic agents. Among the enzymes for which sulfamide-based inhibitors were designed, are the carbonic anhydrases (CAs), a large number of proteases belonging to the aspartic protease (HIV-1 protease, gamma-secretase), serine protease (elastase, chymase, tryptase, and thrombin among others), and metalloprotease (carboxypeptidase A (CPA) and matrix metalloproteinases (MMP)) families. Some steroid sulfatase (STS) and protein tyrosine phosphatase inhibitors belonging to the sulfamide class of derivatives have also been reported. In all these compounds, many of which show low nanomolar affinity for the target enzymes for which they have been designed, the free or substituted sulfamide moiety plays important roles for the binding of the inhibitor to the active site cavity, either by directly coordinating to a metal ion found in some metalloenzymes (CAs, CPA, STS), usually by means of one of the nitrogen atoms present in the sulfamide motif, or as in the case of the cyclic sulfamides acting as HIV protease inhibitors, interacting with the catalytically critical aspartic acid residues of the active site by means of an oxygen atom belonging to the HN-SO2-NH motif, which substitutes a catalytically essential water molecule. In other cases, the sulfamide moiety is important for inducing desired physico-chemical properties to the drug-like compounds incorporating it, such as enhanced water solubility, better bioavailability, etc., because of the intrinsic properties of this highly polarized moiety when attached to an organic scaffold. This interesting motif is thus of great value for the design of pharmacological agents with a lot of applications. PMID- 16710860 TI - New insights on the anticancer properties of dietary polyphenols. AB - Cancer, one of the major causes of death across the world, has shown to be a largely preventable disease, highly susceptible to modulation by dietary factors. Phenolic compounds, abundant in vegetables and fruits ubiquitous in diet, were described to play an important role as chemopreventive agents. Since conventional therapeutic and surgical approaches have not been able to control the incidence of most cancer types, the development of chemopreventive strategies is an urgent priority in public health. The current diet phenolic intake is often insufficient to protect from mutagens (either exogenous or endogenous), which leads to the need for dietary supplementation as an alternative approach. Research efforts are placing increasing emphasis on identifying the biological mechanisms and in particular the signal transduction pathways related to the chemopreventive activities of these compounds. These effects are believed to occur by the regulation of signaling pathways such as nuclear factor-kappaB (NF-kappaB), activator protein-1 (AP-1) or mitogen-activated protein kinases (MAPK). Dietary polyphenols can exert their effects on these pathways separately or sequentially and in addition the occurrence of crosstalk between these pathways cannot be overlooked. By modulating cell signaling pathways, polyphenols activate cell death signals and induce apoptosis in precancerous or malignant cells resulting in the inhibition of cancer development or progression. However, regulation of cell signaling pathways by dietary polyphenols can also lead to cell proliferation/survival or inflammatory responses due to increased expression of several genes. The present review summarizes the most recent advances providing new insights into the molecular mechanisms underlying the promising anticarcinogenic activity of dietary polyphenols. PMID- 16710861 TI - Matrix metalloproteinase 11 (MMP-11; stromelysin-3) and synthetic inhibitors. AB - Matrix metalloproteinase (MMP)-11, or Stromelysin 3, is a particular member of MMP family, a group of zinc-dependent endopeptidases involved in matrix degradation and tissue remodeling. Despite intense efforts since its first characterization 15 years ago, its role and target substrates in different diseases remain largely unknown. While mice with MMP-11 deficiency display no particular phenotype, analysis of different tumorigenesis models with these mice lead to the conclusion that MMP-11 promotes tumor development. In contrast with other MMPs, MMP-11 is unable to degrade any major extracellular matrix component and unlike most of other MMPs that are secreted as inactive proenzymes and activated extracellularly, MMP-11 is secreted under active form. MMP-11 may thus play a unique role in tissue remodeling processes, including those associated with tumor progression. Although MMP-11 and other MMPs have been considered as promising targets to combat cancer, a first series of clinical trials using broad spectrum MMP inhibitors have not led to significant therapeutic benefits. These disappointing results highlight the need for better understanding of the exact role played by each MMP during the different stages of tumor progression. Among the different strategies to fill this gap, highly specific MMP inhibitors would be of great value. This review provides an update on the selectivity profile of phosphinic MMP-11 synthetic inhibitors developed and discusses the opportunities and limitations to identify inhibitors able to fully discriminate MMP-11 from the other MMPs. PMID- 16710862 TI - Endometrial receptivity: clinical assessment in relation to fertility, infertility, and antifertility. AB - Fertility in humans and other mammalian species depends absolutely on synchronous events that render the developing blastocyst and the receiving uterus competent for implantation. Endometrial receptivity is defined as the period during which the endometrial epithelium acquires functional, but transient, ovarian steroid dependent status supportive to blastocyst acceptance and implantation. Once inside the uterus, the blastocyst is surrounded by an intact luminal epithelium, which is considered to act as barrier to its attachment, except for this short period of high endometrial receptivity to blastocyst signal(s). Its transport and permeability properties, in conjunction with cellular action of the endometrium and the embryo, have been suggested to influence creation and maintenance of informational and nutritional status of uterine luminal milieu. This period, also termed as the 'window of implantation,' is limited to days 20-24 of menstrual cycle in humans. However, establishment of endometrial receptivity is still a biological mystery that remains unsolved despite marked advances in our understanding of endometrial physiology following extensive research associated with its development and function. This review deals with various structural, biochemical, and molecular events in the endometrium coordinated within the implantation window that constitute essential elements in the repertoire that signifies endometrial receptivity and is aimed to achieve a better understanding of its relationship to fertility, infertility, and for the development of targeted antifertility agents for human use and welfare. PMID- 16710863 TI - The reaction of o-alkynylarene and heteroarene carboxaldehyde derivatives with iodonium ions and nucleophiles: a versatile and regioselective synthesis of 1H isochromene, naphthalene, indole, benzofuran, and benzothiophene compounds. AB - The reaction of o-alkynylbenzaldehydes 1 with different alcohols, silylated nucleophiles 5, electron-rich arenes 10, and heteroarenes 12 in the presence of the reagent IPy(2)BF(4), at room temperature, gave functionalized 4-iodo-1H isochromenes 2, 6, 11, and 13 in a regioselective manner. When alkynes 16 and alkenes 19 and 20 were used as nucleophiles, a regioselective benzannulation reaction took place to form 1-iodonaphthalenes 17 and 1-naphthyl ketones 18, respectively. Moreover, the latter process has been adapted to accomplish the synthesis of indole, benzofuran, and benzothiophene derivatives (23, 27, and 28, respectively). The three patterns of reactivity observed for the o alkynylbenzaldehyde derivatives with IPy(2)BF(4) stem from a common iodinated isobenzopyrylium ion intermediate, A, that evolves in a different way depending on the nucleophile present in the reaction medium. A mechanism is proposed and the different reaction pathways observed as a function of the type of nucleophile are discussed. Furthermore, the reaction of the o-hexynylbenzaldehyde 1 b with styrene was monitored by NMR spectroscopy. Compound III, a resting state for the common intermediate in the absence of acid, has been isolated. Its evolution in acid media has been also tested, thereby providing support to the proposed mechanism. PMID- 16710864 TI - A QM/MM study of cisplatin-DNA oligonucleotides: from simple models to realistic systems. AB - QM/MM calculations were employed to investigate the role of hydrogen bonding and pi stacking in several single- and double-stranded cisplatin-DNA structures. Computed geometrical parameters reproduce experimental structures of cisplatin and its complex with guanine-phosphate-guanine. Following QM/MM optimisation, single-point DFT calculations allowed estimation of intermolecular forces through atoms in molecules (AIM) analysis. Binding energies of platinated single-strand DNA qualitatively agree with myriad experimental and theoretical studies showing that complexes of guanine are stronger than those of adenine. The topology of all studied complexes confirms that platination strongly affects the stability of both single- and double-stranded DNAs: Pt-N-H...X (X = N or O) interactions are ubiquitous in these complexes and account for over 70 % of all H-bonding interactions. The pi stacking is greatly reduced by both mono- and bifunctional complexation: the former causes a loss of about 3-4 kcal mol(-1), whereas the latter leads to more drastic disruption. The effect of platination on Watson Crick GC is similar to that found in previous studies: major redistribution of energy occurs, but the overall stability is barely affected. The BH&H/AMBER/AIM approach was also used to study platination of a double-stranded DNA octamer d(CCTG*G*TCC)d(GGACCAGG), for which an experimental structure is available. Comparison between theory and experiment is satisfactory, and also reproduces previous DFT-based studies of analogous structures. The effect of platination is similar to that seen in model systems, although the effect on GC pairing was more pronounced. These calculations also reveal weaker, secondary interactions of the form Pt...O and Pt...N, detected in several single- and double-stranded DNA. PMID- 16710865 TI - Triply fused Zn(II)-porphyrin oligomers: synthesis, properties, and supramolecular interactions with single-walled carbon nanotubes (SWNTs). AB - The photophysical, electrochemical, and self-assembly properties of a novel triply fused Zn(II)-porphyrin trimer were investigated and compared to the properties of a triply fused porphyrin dimer and the analogous monomer. The trimer exhibited significantly red-shifted absorption bands relative to the corresponding monomer and dimer. Electrochemical investigations indicated a clear trend in redox properties amongst the three porphyrin structures, with the lowest oxidation potential and the lowest HOMO-LUMO gap exhibited by the triply fused trimer. This electrochemical behavior is attributed to the extensive pi-electron delocalization in the trimeric structure relative to the monomer and dimer. Additionally, it was found that the trimer forms extremely strong and nearly irreversible supramolecular interactions with single-walled carbon nanotubes (SWNTs), resulting in stable solutions of porphyrin-nanotube complexes in THF. Formation of these complexes required the addition of trifluoroacetic acid (TFA) to the solvent. This allowed the oligomers to make close contact with the nanotubes, enabling the formation of stable supramolecular assemblies. Atomic force microscopy (AFM) was used to observe the supramolecular porphyrin-nanotube complexes and revealed that the porphyrin trimer formed a uniform coating on the SWNTs. Height profiles indicated that nanotube bundles could be exfoliated into either individual tubes or very small bundles by exposure to the porphyrin trimer during sonication. PMID- 16710866 TI - Modular approach to tridentate N,O,N' ligands using pyrazolylborate chemistry. AB - Two anionic tridentate N,O,N' chelators, [pz(Ph)B(mu-pz)(mu-O)B(Ph)pz](-) (3(-)) and [pz(Ph)(Ph)B(mu-pz)(mu-O)B(Ph)pz(Ph)](-) (4(-)), as well as the corresponding complexes [Fe(3)(py)Cl], [Fe(3)Cl(2)] and [Cu(3)Cl], have been synthesised and structurally characterised by X-ray crystallography (pz: pyrazolyl, pz(Ph): 3 phenylpyrazolyl, py: pyridine). Since our synthesis approach takes advantage of the highly modular pyrazolylborate chemistry, inexpensive and relatively resistant N,O,N' ligands of varying steric demand are readily accessible. The complexes [Fe(3)(py)Cl] and [Fe(3)Cl(2)] possess a distorted trigonal-bipyramidal configuration with the pyrazolyl rings occupying equatorial positions and the oxygen donor being located at an apical position. The complex [Cu(3)Cl] crystallises as chloro-bridged dimers featuring Cu(II) ions with ligand environments that are intermediate between a square-planar and a trigonal bipyramidal geometry. PMID- 16710867 TI - Toward the synthesis of the antibiotic branimycin: novel approaches to highly substituted cis-decalin systems. AB - A variety of highly functionalized cis-decalin systems have been prepared by means of the stereoselective transannular Diels-Alder (TADA) reaction of a (Z,E,Z,Z)-tetraene macrolide, and by means of intramolecular nitrile oxide olefin (INOC) or ring-closing metathesis (RCM) annulations to quinic acid derivatives. PMID- 16710869 TI - Cyclic oligomers of oxetane-based dipeptide isosteres derived from L-rhamnose. AB - Two new cyclic oligomers, cyclo-tetra-[2,4-anhydro-3-O-tert-butyldimethylsilyl-5 deoxy-L-rhamnonamido-(N-->5)] and the corresponding 6-deoxy-D-gulonate cyclic "tetramer", have been synthesised from linear tetrameric oligomers, using TBTU- and pentafluorophenyl ester-based methodologies, respectively. These two compounds constitute a novel class of cyclic oligomers derived from oxetane-based sugar amino acids. PMID- 16710868 TI - Synthesis of size-controlled acid-resistant hybrid calcium carbonate microparticles as templates for fabricating "micelles-enhanced" polyelectrolyte capsules by the LBL technique. AB - Size-controlled, low-dispersed calcium carbonate microparticles were synthesized in the presence of the amphiphilic block copolymer polystyrene-b-poly(acrylic acid) (PS-b-PAA) by modulating the concentration of block copolymer in the reactive system. This type of hybrid microparticles have acid-resistant properties. By investigating the aggregation behaviors of PS-b-PAA micelles by transmission electron microscopy (TEM), the mechanism of hybrid calcium carbonate formation illustrated that the block copolymer served not only as "pseudonuclei" for the growth of calcium carbonate nanocrystals, but also forms the supramicelle congeries, a spherical framework, as templates for calcium carbonate nanocrystal growth into hybrid CaCO(3) particles. Moreover, this pilot study shows that the hybrid microparticle is a novel candidate as a template for fabricating multilayer polyelectrolyte capsules, in which the block copolymer is retained within the capsule interior after core removal under soft conditions. This not only facilitates the encapsulation of special materials, but also provides "micelles-enhanced" polyelectrolyte capsules. PMID- 16710870 TI - Efficient methodology for the cyclization of linear peptide libraries via intramolecular S-alkylation using Multipin solid phase peptide synthesis. AB - Methodology is described here for the efficient parallel synthesis and cyclization of linear peptide libraries using intramolecular S-alkylation chemistry in combination with Multipin solid phase peptide synthesis (Multipin SPPS). The effective use of this methodology was demonstrated with the synthesis of a 72-member combinatorial library of cyclic thioether peptide derivatives of the conserved four-residue structural motif DD/EXK found in the active sites of the five crystallographically defined orthodox type II restriction endonucleases, EcoRV, EcoRI, PvuII, BamHI and BglI. PMID- 16710871 TI - Peptide vector for gene delivery with high affinity for phosphatidylserine. AB - Since phosphatidylserine (PS) is known to translocate to the external face of the plasma membrane when the cell membrane becomes disordered, we decided to focus our attention on PS as a target molecule for gene delivery. In this paper, the novel peptide Td3701 was designed, synthesized, and characterized for its physico chemico-biological properties. Td3701 simultaneously exhibited both characters as a DNA carrier and a sensor probe for active targeting, which seemed to be triggered by structural changes in the presence of PS. This is a very unique character among nonviral vectors, and it is believed that Td3701 could be used for selective gene delivery. PMID- 16710873 TI - Greener media in chemical synthesis and processing. PMID- 16710872 TI - Peptide chemistry at Oxford before the Second World War. AB - The largely unnoticed and unpublished peptide science interests of Nobel Laureate Sir Robert Robinson at Oxford during the period 1936-1939 are outlined. PMID- 16710874 TI - Building complex glycopeptides: Development of a cysteine-free native chemical ligation protocol. PMID- 16710875 TI - Solid-phase oligosaccharide tagging (SPOT): Validation on glycolipid-derived structures. PMID- 16710876 TI - Pericyclic cascade reactions of (bicyclo[1.1.0]butylmethyl)amines. PMID- 16710877 TI - Normal fetal motility: an overview. AB - After 35 years of real-time two-dimensional sonography, and now that 4D sonography is within our grasp, this article presents an overview of present-day knowledge of normal fetal motility. A literature search was carried out on articles from 1970, using the keywords: 'fetal', 'movements', 'motility', 'movement patterns', 'ultrasound' and 'sonography'. Inclusion criteria were human studies and use of real-time sonography. Articles were screened for type of motor assessment procedure, in terms of whether they: specified movements for participating body parts (specific movement pattern, SMP), were qualitative (performance in terms of speed and amplitude), were quantitative, identified behavioral states, stated the duration of observation, and specified gestational age. We noted developmental milestones obtained for each study aim. One of four aims was identified for each article, depending on whether it focused on emergence, development, or continuity after birth of the movement patterns, or on the relationship of various motor aspects to other parameters that evaluate fetal condition, such as blood flow and fetal heart rate. A total of 109 relevant articles was identified, examining 9862 fetuses. Assessment was performed primarily with analysis of SMPs (89%); 52% also included non-SMPs (NSMPs), 78% included quantification, 24% assessment of quality, and 32% behavioral states. The duration of observation was 1 h or longer in 50% of the studies. The focus in 28 studies was on emergence, in 44 it was on development, in five it was on continuity and in 32 it was on relationship of the movements with other parameters of fetal well-being. A few milestones identified were determination of the strictly age-related emergence of SMPs and behavioral states, the highly reproducible quality of SMPs throughout gestation, the age-related trends in quantified SMPs, the continuity in quality and quantity after birth, and the close relationship between motility and heart-rate variability, flow parameters, and behavioral states. Periods of longest inactivity recorded before 20 weeks were 13 min; after 30 weeks they were 45 min. Much insight was obtained into the development of motility and its relationship to other parameters from those articles applying comparable assessment procedures. An assessment procedure with well-defined SMPs, qualitative and quantitative aspects of SMPs and NSMPs, and an observation period dependent on age are advocated for future research. PMID- 16710878 TI - Risk factors for tumour present in a circumferential line of resection after excision of rectal cancer. AB - BACKGROUND: Transected tumour in a circumferential line of resection after excision of rectal cancer carries a high likelihood of local recurrence. The aim of this study was to identify independent risk factors for transected tumour and to examine their temporal variability. METHODS: Data were drawn from a comprehensive, prospective hospital registry of all resections for rectal cancer from January 1971 to July 2004. Transected tumour was defined as tumour present histologically in a line of resection and was assessed in all specimens. RESULTS: Transection occurred in 129 of 1613 patients (8.0 (95 per cent confidence interval 6.7 to 9.4) per cent). The following variables were independently associated with transected tumour: tumour perforation, a non-restorative operation, tumour adherence, non-standardized operative technique, preoperative radiotherapy, male sex, histological involvement of an adjacent organ or tissue, high-grade tumour and venous invasion. The mean number of risk factors per patient per year and the annual percentage of patients with transection varied distinctly over the history of the database. CONCLUSION: The varying prevalence of risk factors, both within and between hospitals and patient series, should be taken into account if the rate of transection is to be regarded as an index of the quality of surgery. PMID- 16710879 TI - Ultrasonographic characteristics and cystoscopic correlates of bladder wall invasion by endophytic cervical cancer. AB - OBJECTIVES: To present the ultrasonographic findings and explore cystoscopic correlates of endophytic cervical cancer invading the bladder wall. METHODS: We retrospectively reviewed the imaging files in 19 cases of cervical cancer, comprising six cases of endophytic lesions invading the bladder wall (study group) and 13 cases of endo- or exophytic, clinical stage IB1 lesions without bladder wall invasion (controls). Bladder wall infiltration or invasion was confirmed by cystoscopic biopsy or surgical findings or both. All 19 patients had undergone transvaginal ultrasound examination to evaluate the cervix and lower urinary tract. The main measures included tumor volume of the primary cervical neoplasm, tumor protuberance, mobility of the cervix over the lower bladder wall, continuity of the endopelvic fascia and echogenicity and morphological texture of the bladder wall and its thickness. RESULTS: A tumor protuberance emerging from the cervicocorporeal junction and invading the bladder in the supratrigonal area was seen in the study group but not in the controls. Disruption of the endopelvic fascia, a thickened bladder wall, changes in the bladder mucosa and interruption of the entire bladder wall were ultrasonographic characteristics demonstrating the sequential stages of bladder wall invasion. The morphological changes in the bladder wall on ultrasound examination were categorized into four stages. Based on the morphological classification, two of the six patients in the study group belonged to stage I, one to stage II, one to stage III and two to stage IV of bladder wall invasion. CONCLUSIONS: In cervical cancer, transvaginal ultrasonography helps to explore sequential changes seen in bladder wall invasion, information that is not as readily available from cystoscopic examination. PMID- 16710880 TI - Systematic review of postoperative complications in patients with inflammatory bowel disease treated with immunomodulators. AB - BACKGROUND: This systematic review examined the use of immunomodulators and the risk of postoperative complications after abdominal surgery in patients with inflammatory bowel disease. METHODS: Electronic databases (PubMed, Embase, Ingenta, Zetoc and Ovid) were searched and the reference lists in all articles identified were hand-searched for further relevant papers. Studies were included if they evaluated postoperative complications and defined exposure to individual immunomodulators. RESULTS: All 11 studies that met the inclusion criteria were observational studies; two were reported only in abstract form. Five studies reported risks associated with azathioprine, five reported risks associated with cyclosporin and three reported risks associated with infliximab. None showed an increased risk of either total or infectious complications associated with immunomodulator use. However, subgroup analysis in one study, published as an abstract, suggested increased rates of anastomotic complications and reoperation associated with azathioprine. CONCLUSION: Available evidence does not suggest an increased rate of postoperative complications associated with immunomodulator use. PMID- 16710881 TI - Prenatal echocardiography in a case of Uhl's anomaly. PMID- 16710882 TI - Three-dimensional ultrasound volume calculations of human embryos and young fetuses: a study on the volumetry of compound structures and its reproducibility. AB - OBJECTIVE: To evaluate volumetry with three-dimensional (3D) ultrasonography in the assessment of the size of human embryos and fetuses. METHODS: Forty-four healthy embryos/fetuses with crown-rump length (CRL) ranging from 9 mm to 58 mm were studied using a 7.5-MHz annular array transvaginal 3D probe. EchoPAC 3D software was used to calculate the volumes of the head, body and limbs in the same data set by two observers working independently of each other. Regression analysis was used to assess the relationship between estimated volumes and CRL. RESULTS: The embryonic and fetal volume estimates of both observers ranged from a mean of 93 mm3 at 10 mm CRL to a mean of 11 169 mm3 at 55 mm CRL. The volume of the limbs as a proportion of the mean whole-body volume increased from 4.7% at a CRL of 15 mm to 9.3% at a CRL of 55 mm. Limits of agreement between the observers were calculated to be -0.12 +/- 9.2%. CONCLUSION: It is possible to reconstruct complex small anatomic structures and calculate the volumes of human embryos and fetuses in vivo by using dedicated 3D ultrasound equipment. The reproducibility of whole-body volume estimates seems to be high. The limbs represent a significant proportion of the size of the embryonic/fetal body. PMID- 16710883 TI - Visibility of the polypropylene tape after tension-free vaginal tape (TVT) procedure in women with stress urinary incontinence: comparison of introital ultrasound and magnetic resonance imaging in vitro and in vivo. AB - OBJECTIVE: To determine whether introital sonography and magnetic resonance imaging (MRI) after TVT (tension-free vaginal tape) insertion can depict the polypropylene tape, and thus be used for patient follow-up. METHODS: The study comprised an experimental part, which investigated in-vitro visualization of the polypropylene tape in a model (phantom), and a clinical part, in which 20 women (mean age, 53.4 years) with clinically and urodynamically proven stress urinary incontinence without prolapse were investigated by introital ultrasound and MRI before and 13 months after the TVT procedure. RESULTS: In the phantom, the polypropylene tape was depicted with a low signal intensity by MRI and as a highly echogenic structure by ultrasound. In the clinical study, introital ultrasound in a mediosagittal orientation depicted the vaginal tape in all patients: it was located under either the midurethra (n = 16) or the lower urethra (n = 4), and in either the muscular coat of the urethra (n = 8) or in the urethrovaginal space (n = 12), the tape was either flat (n = 6) or curled up (n = 14), and there was no retropubic visualization of the tape. Overall, depiction by MRI was limited, and was poorer in comparison with ultrasound, especially when the tape had a sub- or paraurethral location. Retropubically, however, MRI identified the tape near the periosteum of the pubic bone (55% of cases), in the retropubic space (37.5% of cases), or near the bladder wall (7.5% of cases). CONCLUSION: Sonography is recommended for evaluation of the suburethral and paraurethral tape portions, while MRI is suitable for retropubic evaluation after the TVT procedure. PMID- 16710884 TI - Sequential triage in the first trimester may enhance advanced ultrasound scanning in population screening for trisomy 21. AB - OBJECTIVE: To design a trisomy 21 screening protocol for sequential triage in the first trimester, and to evaluate whether it reduces the need for advanced ultrasound scanning to such an extent that this could be dealt with by a limited number of well-trained sonographers only. METHODS: Screening results of 31 trisomy 21 affected pregnancies and 16 096 unaffected pregnancies from the first trimester screening program of Algemeen Medisch Laboratorium in Antwerp, Belgium, were used to define high-risk, intermediate-risk and low-risk groups. A serum screening result (age, pregnancy-associated plasma protein-A (PAPP-A) and free beta-human chorionic gonadotropin (beta-hCG)) of >or=1 : 30 and/or a nuchal translucency thickness (NT) measurement of >or= 3.5 mm were classified as high risk. A serum screening result of < 1 : 1000 together with an NT of < 3.5 mm were classified as low risk. Other results were considered intermediate risk, for which further advanced ultrasound screening would be indicated. This protocol was then evaluated prospectively in another population of 13 493 first-trimester pregnancies. RESULTS: Of the total population, 1.9% was identified as being high risk (14 trisomy 21 pregnancies and 222 unaffected pregnancies; prevalence, 1 : 17), 59.6% was identified as being low risk (three trisomy 21 pregnancies and 9615 unaffected pregnancies; prevalence, 1 : 3206) and 38.4% was identified as being intermediate risk (10 trisomy 21 pregnancies and 6190 unaffected pregnancies; prevalence, 1 : 620). A similar distribution was found in the prospective arm of the study. There was no reduction of overall screening performance compared with our current first-trimester combined screening program. The number of intermediate-risk pregnancies was sufficiently low as to enable advanced ultrasound scanning by well-trained sonographers only. CONCLUSION: In population screening for fetal trisomy 21, sequential triage in the first trimester can be achieved using very simple methods. Pregnancies at high or at low risk can be identified easily and the number of pregnancies at intermediate risk can be reduced sufficiently to enable advanced ultrasound scanning by well trained sonographers only. A prospective study is needed to evaluate the performance of this approach and to compare its results with current combined or integrated screening algorithms. PMID- 16710885 TI - Two- and three-dimensional ultrasonographic findings in urethral stenosis with bladder wall trabeculation: case report. AB - Female bladder outlet obstruction is uncommon. We report a case of bladder outlet obstruction secondary to urethral stenosis leading to bladder wall trabeculation. The patient presented at our clinic because of lower urinary tract symptoms including nocturia, urgency, bed wetting, hesitancy, straining to void, and incomplete emptying. Urodynamic study revealed a low maximum free uroflow rate, high residual urine volume, and low compliance on filling phase cystometry. Introital ultrasonography with two- and three-dimensional (2D and 3D) scanning displayed a constriction in the echolucent part of the lower-mid urethra with hyperechogenicity and a lattice-like appearance of the bladder wall. Cystourethroscopy confirmed urethral stenosis and bladder wall trabeculation. The voiding symptoms subsided after the urethral stenosis was relieved by urethral dilatation, but, despite some improvement, the irritative symptoms persisted. Introital ultrasonography with 2D and 3D scanning may help to clarify the cause of female bladder outlet obstruction. PMID- 16710886 TI - Changes in levels of serum insulin, C-Peptide and glucose after electroacupuncture and diet therapy in obese women. AB - Our purpose was to investigate the effects of electroacupuncture (EA) therapy on body weight and on levels of serum insulin, c-peptide and glucose in obese women. 52 healthy women were included in this study and were allocated into three groups: 1) Placebo EA group (n = 15; mean age = 41.8 +/- 4.6 and mean body mass index {BMI} = 33.2 +/- 3.5); 2) EA group (n = 20; mean age = 42.1 +/- 4.4 and BMI = 35.9 +/- 3.6) and 3) Diet restriction group (n = 20; mean age = 42.9 +/- 4.3 and BMI = 34.7 +/- 2.7). EA was applied to the ear points Hunger and Shen Men on alternating days and to the body points LI 4, LI 11, St 36 and St 44 once a day for 30 minutes over 20 days. Diet restriction that entailed a 1450 kilocalorie (kcal) diet program was applied to the three groups for 20 days. An increase in weight loss was observed when weight loss in the EA group (p < 0.000) was compared to that in the diet restricted and placebo EA groups using the Tukey HSD test. There were increases in the serum insulin (p < 0.001) and c-peptide levels (p < 0.000) in the women treated with EA compared to those in the women treated with the placebo EA and diet restriction groups. A decrease was observed in the glucose levels (p < 0.01) in both the EA and diet restriction groups compared to those in the placebo EA group. Our results suggest that EA therapy is an effective method in treating obesity. EA therapy also helps serum glucose levels to decrease through the increase of serum insulin and c-peptide levels. PMID- 16710887 TI - Differential gene expression in hemodialysis patients with "cold" zheng. AB - The aim of the present study was to search for the differential gene expression and measure the serum level of a number of biochemical parameters in the cold zheng (CZ) and non-cold zheng (NCZ) in patients receiving hemodialysis. Hemodialysis (HD) patients were randomly selected from the CZ and NCZ groups. The between-group differences in gene expression were assessed using complementary DNA (cDNA) microarray. Differential gene expression was further validated by real time reverse transcriptase polymerase chain reaction (RT-PCR). Our results demonstrated that the up-regulation of the inflammation-associated genes, ALOX5AP, S100A8 and S100A12, down-regulation of the genes related to immunity (DEFA4), metabolism (GNG11, PYGB, PRKAR2B), and growth/proliferation (HSF2, DDR2, TK1) were found in the CZ group. Furthermore, the CZ HD patients had significantly lower serum albumin levels compared with their NCZ counterparts (3.31 +/- 0.08 g/dL versus 4.18 +/- 0.12 g/dL). It appears reasonable to conclude that up-regulated inflammatory-gene expression (ALOX5AP, S100A8 and S100A12) may play an important role in CZ HD patients. PMID- 16710889 TI - Pinelliae Rhizoma herbal-acupuncture solution induced apoptosis in human cervical cancer cells, SNU-17. AB - Pinelliae Rhizoma has been used traditionally in Korea to promote the liver Qi activity and the function of the digestive system. We investigated whether the Pinelliae Rhizoma herbal-acupuncture solution (PRHS) would induce cell-death on SNU-17, human cervical cancer cells. 3-(4,5-Dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay was used to investigate the cytotoxicity of PRHS. The cell death was identified as apoptosis with 4, 6-diamidineo-2 phenylindole (DAPI) staining, and terminal deoxy-nucleotidyl transferase (TdT) mediated dUTP nick end labeling (TUNEL) assay. PRHS could induce apoptosis of SNU 17 via Bax-related caspase-3 activation. The expressions of both Bax, a pro apoptotic gene, and caspase-3, an apoptotic gene, were increased. The results might provide the experimental data for the clinical use of Pinelliae Rhizoma on cervical cancer. PMID- 16710888 TI - Clinical and molecular evaluation of warming and tonic herb treatment for sibling patients of a typical kidney-yang deficiency family. AB - It is essential to explore the molecular therapeutic effect of warm and tonic herb treatment for individuals with typical kidney-yang deficiency. In this report, we have identified members of a family with a history of suffering from cold and kidney-yang deficiency syndromes. First, we have employed the accumulated scores of the 40-items clinical scoring indicators for kidney-yang deficiency and cold syndromes to clinically assess the presence or absence of the deficiency for 15 family members. We then proceeded to compare the gene expression profiles of RNA isolated from blood samples, prior to and post-herbal treatment, of a sibling (brother and younger sister) that are suffering from the deficiency using cDNA microarrays with 18816 genes. Following treatments with the warming and tonic herb, the accumulated clinical scores obtained from the 40 items clinical scoring indicators were compared to those obtained pre-treatment. It was observed that the accumulated clinical scores were reduced by 1/3 for the brother and 2/3 for his younger sister following the treatments. Furthermore, we have demonstrated that the level of gene expression for a total of 33 genes at pre-treatment was modulated after treatments with the warming and tonic herb and correlated well with the clinical improvements of their syndromes. These results suggest that the combination of gene profiling and the accumulated clinical scores obtained from the 40-items clinical scoring indicators may provide an accurate clinical assessment and a way to monitor the therapeutic efficacy of the warming and tonic herb treatment. PMID- 16710890 TI - Acupuncture for diagnostic fiberoptic bronchoscopy: a prospective, randomized, placebo-controlled study. AB - Patients usually fear fiberoptic bronchoscopy (FBS) and they report a low level of satisfaction after this examination. We evaluated the efficacy of acupuncture in decreasing patient anxiety before diagnostic FBS and in improving tolerance to the examination. In a prospective double-blind study, we enrolled 48 patients scheduled to undergo diagnostic FBS. Patients were randomly assigned to one of three groups. Group A (16 patients): standard FBS, with airway topic anesthesia; Group B (16 patients): standard FBS, with airway topic anesthesia and acupuncture treatment; Group C (16 patients): standard FBS, with airway topic anesthesia and sham acupuncture. EKG, non-invasive arterial pressure, and pulse oximetry were monitored on a routine basis. We evaluated patient anxiety before and after acupuncture and, at the end of FBS, the discomfort suffered during the examination by a 100-mm Visual Analog Scale (VAS). Patient satisfaction in Group A was 50% worse than in Group B (p = 0.04). We observed a strong, even if not statistically significant, tendency toward a lower pre-FBS anxiety in Group B. Patients in group C had values very close to those recorded in group A. We observed no adverse event and no differences in cardio-respiratory parameters in these three groups; in particular, we did not observe a respiratory depression in Group B. Acupuncture seems an effective resource for a Thoracic Endoscopic Room to improve patient tolerance to FBS. PMID- 16710891 TI - Effects of electroacupuncture with different frequencies on the glycoconjugate alterations in articular cartilage in the ankle joints of complete Freund's adjuvant-injected rats. AB - The aim of this study was to investigate the effects of electroacupuncture (EA) on the glycoconjugate (GC) changes in articular cartilage in the ankle of an arthritic model. Arthritis was induced by an intraplantar injection of complete Freund's adjuvant (CFA) into the hindpaw of male Sprague-Dawley rats. Bilateral EA stimulation at 2 Hz, 15 Hz and 120 Hz was applied at those acupoints corresponding to Zusanli and Sanyinjiao in man, using needles for 3-day intervals for 30 days. To determine the presence of arthritis, paw edema was measured by a water displacement plethysmometer. Edema of the hindpaw induced by CFA-injection was strongly inhibited by EA stimulation throughout the experimental period. At 30 days after CFA-injection, GC changes of articular cartilage of the ankle joint were observed using conventional and lectin histochemistry. The CFA-injected rat revealed general reduction of staining abilities and lectin affinities for GC in comparison with normal rats. Significant reductions of neutral and acidic GC were observed in interterritorial matrix and chondrocyte capsules, respectively. All lectin affinities examined except DBA were also decreased in CFA-injected rats compared to normal ones. However, EA-treated rats, showed similar staining patterns and lectin affinities for GC as to normal ones, especially neutral GC in interterritorial matrix and sWGA and RCA-1 affinities in chondrocytes. It is concluded that EA in all frequencies examined, especially 2 Hz, can attenuate inflammatory edema in CFA-injected rats through alleviation of alterations of GC components in articular cartilage. PMID- 16710892 TI - Neurons in the NTS of rat response to gastric distention stimulation and acupuncture at body surface points. AB - The nucleus of the solitary tract (NTS) is a primary center, not only for receiving visceral afferents, but also for somatic afferents. Previous experiments demonstrated that NTS might be a primary afferent center for connecting the Stomach Meridian (ST Medirian) and the Stomach. In this study, an extracellular recording method was used to analyze the firing neurons. The variety of NTS' neurons' responses to gastric distention (GD) and different body surface points were compared. In 52 rats, 104 firing neurons of NTS were recorded, in which there were 47 GD related neurons. For the 47 GD related neurons, there were 33 neurons responsive to acupuncture at sibai (ST 2, belonging to ST Meridian). Therefore, in the 47 GD related neurons, the response neuron occurring rate (RNOR) for acupuncture at ST 2 was 70.21%. Meanwhile in the 47 GD related neurons, when acupuncture was performed at Quanliao (SI 18, belonging to Small Intestine Meridian) and non-acupoint respectively, there were 18 and 17 firing neurons, and the RNOR were 38.30% and 36.17% respectively. The experiment demonstrated that there were some varieties in the response of NTS neurons to GD stimuli and acupuncture at different body surface points, which suggested that the ST Meridian was more intimately linked to the stomach and that the different acupoints do have some specificity. PMID- 16710893 TI - Changes of metallothionein 1 and 3 mRNA levels with age in brain of senescence accelerated mice and the effects of acupuncture. AB - The effects of aging and acupuncture on brain MT1 and MT3 mRNA levels in senescence-accelerated mice (SAMP10) and accelerated senescence resistant mice (SAMR1) were analyzed by Northern blot analysis. Both MT1 and MT3 mRNA levels in SAMR1 were increased significantly from birth to month 4 and decreased gradually thereafter. In SAMP10, the MT3 mRNA level followed the same pattern as in SAMR1 before month 4, then decreased from month 4 to 6, but was over-expressed and exceeded the previous level at month 8. The MT1 mRNA expression in SAMP10 showed a zigzag pattern. Of two groups of SAMP10 mice treated with acupuncture, the xingnao group (PC6 and Du26 as acupoints) and the zibuganshen group (BL18 and BL23 as acupoints), both showed a higher MT1 mRNA level and a lower MT3 mRNA level than the age-matched control group. Meanwhile, in both of the acupuncture groups, the ratios of MT3 to MT1 were down-regulated to the normal range. Overall, these results suggested that over-expression of MT3 mRNA and the increase in MT3 to MT1 ratios in SAMP10 were correlated with aging, and could be an important physiological and pathological event in the aging process. Acupuncture altered the expression levels of MT1 and MT3 mRNA and differences between the effects of the two stimulated acupoints were seen. Therefore, maintenance of the balance between MTs mRNA expression and correct MTs concentrations is crucial for brain-endocrine-immune response and normal aging. Acupuncture could improve zinc ion bioavailability, by maintaining the balance between MT1 and MT3 mRNA expression levels and might explain one of the mechanisms by which acupuncture treatments defer aging and treat some age-related neurodegenerative diseases. PMID- 16710894 TI - The study of dynamic response to acute hemorrhage by pulse spectrum analysis. AB - Traditional Chinese Medicine (TCM) holds that blood and qi are fundamental substances in the human body for sustaining normal vital activity. The theory of qi, blood and zang-fu contribute the most important theoretical basis of human physiology in TCM. An animal model using conscious rats was employed in this study to further comprehend how organisms survive during acute hemorrhage by maintaining the functionalities of qi and blood through dynamically regulating visceral physiological conditions. Pulse waves of arterial blood pressure before and after the hemorrhage were taken in parallel to pulse spectrum analysis. Percentage differences of mean arterial blood pressure and harmonics were recorded in subsequent 5-minute intervals following the hemorrhage. Data were analyzed using a one-way analysis of variance (ANOVA) with Duncan's test for pairwise comparisons. Results showed that, within 30 minutes following the onset of acute hemorrhage,the reduction of mean arterial blood pressure was improved from 62% to 20%. Throughout the process, changes to the pulse spectrum appeared to result in a new balance over time. The percentage differences of the second and third harmonics, which were related to kidney and spleen, both increased significantly than baseline and towards another steady state. Apart from the steady state resulting from the previous stage, the percentage difference of the 4th harmonic decreased significantly to another steady state. The observed change could be attributed to the induction of functional qi, and is a result of qi blood balancing activity that organisms hold to survive against acute bleeding. PMID- 16710895 TI - Pharmacological effects of ba-wei-xi-lei powder on ulcerative colitis in rats with enema application. AB - Ba-wei-xi-lei powder is a classical herbal mixture, and is widely used for the treatment of oral ulcer and ulcerative colitis. This study aimed to explore the effect of Ba-wei-xi-lei powder with enema application on ulcerative colitis in rats. Ulcerative colitis was induced by immunization with rabbit's colonic mucosal protein emulsified with Completely Freund's Adjuvant. The mucosal inflammatory reaction and ulcer have been observed in the model rats. Characteristic changes of ulceractive colitis include that CD4 lymphocyte increased in peripheral blood while CD8 lymphocyte decreased; CD8 lymphocyte and TNF-alpha expression area increased in colonic mucosa, while CD4 lymphocyte decreased. Ba-wei-xi-lei powder and sulfasalazine with enema application could alleviate the pathological changes in the model rats. The results suggest that the pharmacological effects of Ba-wei-xi-lei powder on ulcerative colitis in rats are similar to the effect of sulfasalazine. PMID- 16710896 TI - Hepatoprotective effect of Phyllanthus in Taiwan on acute liver damage induced by carbon tetrachloride. AB - The effect of oral administration of Phyllanthus methanolic extracts (PME) (i.e. P. acidus, P. emblica, P. myrtifolius, P. multiflorus, P. amarus, P. debilis, P. embergeri, P. hookeri, P. tenellus, P. urinaria L.s. nudicarpus, P. urinaria L.s. urinaria) or gallic acid (GA) on the progression of acute liver damage induced by CCl(4) in rats was examined by morphological and biochemical methods. P. acidus, P. urinaria L.s. urinaria, GA at a dose of 0.5 g/kg, and P. emblica, P. urinaria L.s. nudicarpus at a dose of 1.0 g/kg attenuated CCl(4)-induced increase in serum glutamate-oxalate-transaminase (GOT). P. acidus, P. urinaria L.s. nudicarpus, P. urinaria L.s. urinaria, GA at a dose of 0.5 g/kg, and P. emblica, P. amarus, P. hookeri, P. tenellus at a dose of 1.0 g/kg attenuated CCl(4)-induced increase in serum glutamate-pyruvate-transaminase (GPT). Concurrently, P. acidus, P. multiflorus, P. embergeri, P. hookeri, P. tenellus and P. urinaria L.s. urinaria elevated the activity of liver reduced glutathione peroxidase (GSH-Px). Since the protective effects of P. acidus, P. emblica, P. myrtifolius, P. embergeri, P. urinaria L.s. nudicarpus, P. urinaria L.s. urinaria and GA correlate with a reduction in liver infiltration and focal necrosis observed using histological methods, these data demonstrate that P. acidus and P. urinaria L.s. urinaria are hepatoprotective and antioxidant agents. PMID- 16710897 TI - Protective effect of total flavones of rhododendra on ischemic myocardial injury in rabbits. AB - This study was to investigate the effect of total flavones of rhododendra (TFR) on ischemic myocardial injury in rabbits. Rabbit ischemic myocardial injury was induced by occluding the anterior descent of the left artery (LAD). The ECG was recorded; the plasma creatine kinase (CK), nitric oxide (NO) and endothelin-1 (ET 1) levels were measured using spectrophotometry, Griess method and radioimmunoassay, respectively. The myocardial ischemic size and infarction size were determined by dual staining with Evan's blue and Nitroblue tetrazolium reductionest (N-BT). A typical ECG S-T segment elevation and an increase of plasma CK activity were observed 6 and 24 hours after the induction of ischemia. These changes were inhibited in rabbits treated with either TFR (30, 60 mg/kg) or ginkgo biloba extract (EGB) for 7 days, indicating a protective effect of TFR on ischemic myocardial injury. The myocardial ischemic size and infarction size were 40.7 +/- 3.6% and 36.8 +/- 3.6% respectively in the control group, while TFR (60 mg/kg) pretreatment for 7 days significantly reduced both myocardial ischemic size (32.40 +/- 5.38%, p < 0.05) and infarction size (28.7 +/- 5.8%, p < 0.05). In addition, the occlusion of LAD resulted in an increase of ET-1 and a decrease of NO levels in the plasma, effects that were inhibited by TFR treatment, suggesting a possible mechanism for the protective effect of TFR against myocardial ischemic injury. PMID- 16710898 TI - Cardiac protective effect of Astragalus on viral myocarditis mice: comparison with Perindopril. AB - In clinical practice, Astragali Radix (Astragalus), the root of Astragalus membranaceus Bunge, has been widely applied to treat patients with viral diseases, including viral myocarditis in China. The present study was designed to evaluate the protective effects of Astragalus on the function of sarcoplasmic reticulum calcium ATPase (SERCA2) activity and endothelin system at acute and chronic periods of myocarditis mice induced by CVB(3) infection. Astragalus feeding (2.2 mg/kg/day) could significantly increase the survival rate, alleviate pathological alterations and serum cardiac troponin I (cTnI), as well as restore impaired SERCA activity at the acute stage. Low affinity and capacity of ETR were reversed with Astragalus after the first CVB(3) inoculation up to 7 days and after the second virus inoculation up to 150 days. In the meantime, the contents of cardiac ET-1 and ANP were reduced. Comparison the myocarditis mice treated with Perindopril (0.44 mg/kg/day), an ACE inhibitor, shows that Astragalus achieved a similar effect on survival rate, SERCA2 and ET system. These results indicated that the beneficial effects of Astragalus and Perindopril for treating viral myocarditis might be partly mediated by preserving the functions of SERCA 2 activity and ET system. PMID- 16710899 TI - An extract of Morinda citrifolia interferes with the serum-induced formation of filamentous structures in Candida albicans and inhibits germination of Aspergillus nidulans. AB - An aqueous extract of Morinda citrifolia was shown to interfere with the serum induced morphological conversion of Candida albicans from a cellular yeast to a filamentous form in vitro. The conversion of C. albicans from a cellular yeast to a filamentous form in vivo is associated with pathogenicity. No significant effect on growth in serum-free media was seen at the concentrations used to interfere with the morphological change. The same extract also inhibited the germination of Apergillus nidulans spores. These results demonstrate that M. citrifolia contains a water-soluble component or components that interfere with the morphological conversion of C. albicans and the germination of A. nidulans and may have potential therapeutic value with regard to candidiasis and aspergillosis. PMID- 16710900 TI - Antimicrobial activities of cinnamon oil and cinnamaldehyde from the Chinese medicinal herb Cinnamomum cassia Blume. AB - Both Cinnamomum verum J.S. Presl. and Cinnamomum cassia Blume are collectively called Cortex Cinnamonmi for their medicinal cinnamon bark. Cinnamomum verum is more popular elsewhere in the world, whereas C. cassia is a well known traditional Chinese medicine. An analysis of hydro-distilled Chinese cinnamon oil and pure cinnamaldehyde by gas chromatography/mass spectrometry revealed that cinnamaldehyde is the major component comprising 85% in the essential oil and the purity of cinnamaldehyde in use is high (> 98%). Both oil and pure cinnamaldehyde of C. cassia were equally effective in inhibiting the growth of various isolates of bacteria including Gram-positive (1 isolate, Staphylococcus aureus), and Gram negative (7 isolates, E. coli, Enterobacter aerogenes, Proteus vulgaris, Pseudomonas aeruginosa, Vibrio cholerae, Vibrio parahaemolyticus and Samonella typhymurium), and fungi including yeasts (four species of Candida, C. albicans, C. tropicalis, C. glabrata, and C. krusei), filamentous molds (4 isolates, three Aspergillus spp. and one Fusarium sp.) and dermatophytes (three isolates, Microsporum gypseum, Trichophyton rubrum and T. mentagraphytes). Their minimum inhibition concentrations (MIC) as determined by agar dilution method varied only slightly. The MICs of both oil and cinnamaldehyde for bacteria ranged from 75 microg/ml to 600 microg/ml, for yeasts from 100 microg/ml to 450 microg/ml, for filamentous fungi from 75 microg/ml to 150 microg/ml, and for dermatophytes from 18.8 microg/ml to 37.5 microg/ml. The antimicrobial effectiveness of C. cassia oil and its major constituent is comparable and almost equivalent, which suggests that the broad-spectrum antibiotic activities of C. cassia oil are due to cinnamaldehyde. The relationship between structure and function of the main components of cinnamon oil is also discussed. PMID- 16710901 TI - Inhibition of melanogenesis in murine B16/F10 melanoma cells by Ligusticum sinensis Oliv. AB - Ligusticum sinensis Oliv. (LSO) is an herbal drug commonly used as a topical treatment of epidermal hyperdepigmentation in Chinese medicine. However, the mechanism underlying the depigmentation by LSO is still unclear. The purpose of this study was to investigate the effects of LSO on the process of melanogenesis and its possible underlying mechanism. Suppressed DOPA oxidase activity of mushroom tyrosinase was first noted when incubated with aqueous extracts of LSO, demonstrating the direct inhibitory effect of LSO on mushroom tyrosinase. Further experiments were carried out in murine B16/F10 melanoma cells and the effects of LSO extract on melanin formation, tyrosinase activity and tyrosinase gene expression were tested. Under conditions without affecting the viability of murine B16/F10 melanoma cells, LSO extract significantly reduced the cellular melanin content in a dose-dependent manner. The DOPA oxidase activity of tyrosinase in B16/F10 cells was dose-dependently inhibited by LSO treatment, possibly mediated by the suppressed tyrosinase mRNA expression in LSO-treated B16/F10 cells. In conclusion, the inhibitory effect of LSO on melanogenesis is likely associated with decreased DOPA oxidase activity of tyrosinase that is most likely the result of the down-regulation of tyrosinase mRNA expression. PMID- 16710902 TI - IBM opens the IP vaults. PMID- 16710904 TI - Abstracts of the 4th International Symposium on Myelodysplastic Syndromes in Childhood, April 24-26, 2006, Freiburg, Germany. PMID- 16710903 TI - Why media relations matter. AB - There is a common misperception that only medium- and large-cap biotechs need to understand and interact with the media. As investor demands increase and partnering dynamics evolve, startups need to understand the art of media relations more now than ever before. PMID- 16710905 TI - Glucosamine and chondroitin sulfate for knee osteoarthritis. PMID- 16710906 TI - Glucosamine and chondroitin sulfate for knee osteoarthritis. PMID- 16710907 TI - Cetuximab plus radiotherapy for head and neck cancer. PMID- 16710908 TI - Cetuximab plus radiotherapy for head and neck cancer. PMID- 16710909 TI - Cetuximab plus radiotherapy for head and neck cancer. PMID- 16710911 TI - Mortality after the hospitalization of a spouse. PMID- 16710910 TI - Selective serotonin-reuptake inhibitors and persistent pulmonary hypertension of the newborn. PMID- 16710912 TI - Medicaid: Medicaid: provider reimbursement--2005. End of Year Issue Brief. AB - Since Title XIX of the Social Security Act was enacted in 1965, state Medicaid programs have operated under tight budget constraints. States have recognized that reimbursement rates, whether traditional fee-for-service rates or capitation rates for managed care providers, must be sufficient in order to ensure that Medicaid programs have enough providers to deliver care. However, states have often looked to save money by lowering payments to providers who deliver health care services to Medicaid beneficiaries. This cost crunch has resulted in provider payment rates that are often substantially below market rates. State legislatures, program administrators and providers have sought to find the proper balance between adequate levels of reimbursement and cost control measures. However, dissatisfaction with low reimbursement levels has caused some providers to cease participating in the Medicaid program. This has had a detrimental affect on Medicaid recipients' access to health services. States have become aware of the problem and have tried to revise their rates to find the elusive balance between adequate reimbursement and fiscal control. PMID- 16710913 TI - Drug-related hepatotoxicity. PMID- 16710914 TI - Drug-related hepatotoxicity. PMID- 16710915 TI - Drug-related hepatotoxicity. PMID- 16710916 TI - Drug-related hepatotoxicity. PMID- 16710917 TI - Medicaid: Medicaid: eligibility--2005. End of Year Issue Brief. AB - Medicaid, a joint federal-state program started in 1965 as Title XIX of the Social Security Act, provides health insurance coverage to low-income children, parents meeting specific income thresholds, pregnant women, the elderly and people with disabilities. In 2003, Medicaid provided health care insurance to approximately 40.2 million low-income Americans. That same year, 45 million Americans had no health insurance at all. In order to reduce the number of uninsured people in the United States, state legislators have introduced hundreds of bills over the past decade to expand health insurance coverage to low-income citizens. Health insurance has been extended for low-income children through Medicaid expansions, the State Children's Health Insurance Program (SCHIP) and other medical assistance programs. States, in partnership with the federal government, have also made it easier for disabled individuals to qualify for Medicaid through Ticket-to-Work legislation (which allows individuals with disabilities to remain in the workforce and keep their Medicaid coverage). They also have expanded coverage for senior citizens, especially for their prescription drug needs through Medicaid Pharmacy Plus waivers. In fact, from 1997 to 2003, monthly Medicaid enrollment rose from 31.2 million to 40.6 million recipients. PMID- 16710918 TI - Medicaid: Medicaid: waivers--2005. End of Year Issue Brief. AB - In 1965, President Lyndon Johnson signed the Social Security Act into law, which was designed to help the elderly and the poor receive needed health insurance. This landmark legislation ushered in a new chapter of federal assistance in providing health care, authorizing the creation of both Medicare under Title XVIII and Medicaid under Title XIX. Since January 1966, states have administered medical assistance and medical assistance to the aged programs under Title XIX, with the federal government paying from 50 percent to 85 percent of the cost. When Medicaid was created in 1965 as a joint federal-state medical assistance program, states were initially under strict guidelines to comply with federal requirements in order to receive matching funds from the federal government. However, after years of experience with the program, and with different states having different needs and challenges due to demographics such as size and population, an abundance or paucity of urban areas, age of the population, quantity of providers, and other factors, the federal government eventually allowed "waivers" to the Social Security Act to help states best meet the needs of their Medicaid recipients and to contain costs in their Medicaid programs. In short, the term waiver is used to describe the statutory authority under which the federal government, through the Centers for Medicare and Medicaid Services (CMS), allows states to receive federal matching funds for its programs even though the state is no longer in compliance with technical requirements of the Social Security Act. PMID- 16710919 TI - Commentary: mass media interventions for promoting HIV testing. PMID- 16710920 TI - Health care providers and facilities: medical errors and patient safety--2005. End of Year Issue Brief. PMID- 16710921 TI - Commentary: mass media interventions for promoting HIV testing. PMID- 16710923 TI - You are invited. By Caveman. PMID- 16710922 TI - Health care providers and facilities: nurse workforce--2005. End of Year Issue Brief. PMID- 16710924 TI - Long-term care: assisted living--2005. End of Year Issue Brief. PMID- 16710925 TI - Long-term care: home- and community-based services--2005. End of Year Issue Brief. PMID- 16710926 TI - Long-term care: end of life issues--2005. End of Year Issue Brief. AB - While end-of-life care encompasses many issues, assisted suicide, chronic pain management and advance health care directives have been key aspects of recent legislation. Assisted suicide poses many ethical issues, while advance care directives gained tremendous attention in the wake of the Terri Schiavo case. In addition, states have worked throughout 2005 to evaluate the best means of promoting policies that assist persons who suffer from chronic pain. PMID- 16710927 TI - Pharmaceuticals: pharmacy benefit management regulations--2005. End of Year Issue Brief. PMID- 16710928 TI - Pharmaceuticals: Medicare Modernization Act--2005. End of Year Issue Brief. AB - The enactment of the landmark Medicare Modernization Act of 2003 (MMA) marked the first structural change to Medicare since its inception in 1965. The MMA established the Medicare prescription drug benefit with an intermediate Medicare approved drug discount card program to be implemented six months after enactment and full implementation in January 2006. The MMA marks the first time the federal government would be providing access to prescription drugs for seniors. Though the merits and cost of the MMA continue to be debated, the impending change to how seniors and the low-income access prescription drugs is on the horizon. Historically, state lawmakers believed it was appropriate to provide assistance to the elderly and, in some states, the disabled and indigent, for the purchase of much needed prescription drugs. Since 1975, states have been creating, terminating and redesigning state pharmaceutical assistance programs (SPAPs) that either provides a prescription drug subsidy, discount card program or full benefit design. Currently, 33 states operate pharmaceutical assistance programs that provide coverage for seniors. Now, due to the MMA, the states' role as provider of prescription drug benefits to seniors is being re-evaluated. The MMA is intended to save costs for the states. Medicare beneficiaries, the bulk of enrollees in state pharmaceutical assistance programs, will now access prescription drugs through the new Medicare prescription drug benefit. The MMA mandates a state to federal government cost shift when the financial burden of those who are eligible for both Medicaid and Medicare (dual eligibles) switch to receiving prescription drug coverage through Medicare, with Medicaid only being an option for drugs not covered by Medicare. Employers, many of which have been cutting prescription drug benefits to retirees because of rising health care costs, will now receive subsidies from the federal government to continue providing prescription drug benefits to retirees. Initially, the MMA appeared to be a fiscal reprieve for states and the need for SPAPs limited. As state policymakers began to analyze the MMA's benefit design and regulations, the cost benefit to the states became questionable. There is a great deal of uncertainty for the states about how they should respond in implementing the MMA. As states review final regulations and full implementation draws closer, the financial impact of the Medicare prescription drug benefit and actual benefits offered by Medicare prescription drug plans are unknown. The states' ability to cut costs as a result of the enactment of the Medicare Modernization Act is questionable. This issue brief describes challenges of MMA implementation for the states and provides an overview of current state pharmaceutical assistance programs and 2005 state legislative activity. PMID- 16710929 TI - Health insurance oversight: managed care--2005. End of Year Issue Brief. PMID- 16710930 TI - Oral pathology quiz #50. Case number 2. Dentigerous cyst. PMID- 16710931 TI - Oral pathology quiz #50. Case number 3. Ameloblastoma. PMID- 16710932 TI - Human mesenchymal precursors derived from embryonic stem cells have specialized differentiation potential and are not tumorigenic. PMID- 16710933 TI - High resolution SNP arrays identify amplified regions in lung cancer. PMID- 16710934 TI - Cutting edge matrix assembly: a new high density tissue microarray technology. PMID- 16710935 TI - Regulating ETS transcription factor function by altering a flexible domain. PMID- 16710936 TI - Are p53 post-translational modifications required for its function in vivo? PMID- 16710938 TI - A genome-wide map of active gene promoters. PMID- 16710937 TI - Identification of an amplified transcription factor in melanoma. PMID- 16710939 TI - PGE2 produced by lung cancer suppresses immune function through T-regulatory cells and can be blocked by the COX2 inhibitor Celebrex. PMID- 16710940 TI - Interview with the President-Elect: Dr. L. Stephen Ortego. PMID- 16710941 TI - Your options through the LDA/LDS Endorsed Program. PMID- 16710942 TI - Charting a course to recovery. PMID- 16710943 TI - A view of Hurricanes Katrina and Rita from within the Louisiana dental community. PMID- 16710944 TI - Dental identification after Hurricane Katrina. PMID- 16710945 TI - Statement on tick-borne encephalitis. An Advisory Committee Statement (ACS). PMID- 16710946 TI - Update on influenza vaccination for the 2005-2006 season. An Advisory Committee Statement (ACS). PMID- 16710947 TI - Oxygen delivery comparison of two constant-flow masks during flight to 6863 m. AB - OBJECTIVE: Differences in mask design may alter the oxygen flow required to gain similar oxygenation at a particular altitude. The selection of the most efficient mask would be advantageous for general aviation and other applications where oxygen supply is limited. METHODS: We compared a basic mask (BAS) without valves or oxygen reservoir bag to a test mask (TST) with three valves and a reservoir designed to deliver up to 80% oxygen for critically ill patients. Randomly assigned parachutists (n = 31) used the masks during a flight with a gradual climb to 6863 m (22,500 ft). The oxygen flow was individually controlled to produce oxygen saturation (SpO2) of 95-97% as determined by pulse oximetry. Oxygen flow and SpO2 were obtained every 305 m (1000 ft). RESULTS: Baseline age, lung function indices, and SpO2 were comparable for the two groups. Mean in flight Spo2 values were 95.3 +/- 0.5% for the BAS and 96.2 +/- 1.1 % for the TST, respectively. Above 3965 m (13,000 ft) the TST required significantly less oxygen flow than the BAS to maintain the target SpO2. At 6863 m (22,500 ft), mean oxygen flow was 5.5 +/- 3.5 L x min(-1) for the BAS vs. 3.4 +/- 2.3 L x min(-1) for the TST (p = 0.029). No adverse reactions were reported from either group. CONCLUSION: The TST required significantly less oxygen flow compared with the BAS at high altitudes and may, therefore, reduce total oxygen use, resulting in reduced costs and longer oxygen availability during a flight. PMID- 16710948 TI - [Gene transfection efficiency and destination of recombinant adenovirus gene transfer through internal jugular vein injection in rats]. AB - OBJECTIVE: To investigate the adenovirus-mediated LacZ gene expression and the destination in different organs of SD rats after the intravenous injection in rats. METHODS: Recombinant adenovirus vector containing LacZ was transferred to SD rats by injecting into the internal jugular vein. To identify the sites and periods of LacZ gene expression, X-gal staining was used to detect beta-gal level and period of LacZ gene expression of different organs in the transfected and non transfected rats at different time intervals. RESULTS: On the 1st day after the injection, the lung, liver, kidney, and spleen expressed some beta-gal; on the 3rd day after the injection, the lung, liver, kidney, and spleen expressed beta gal obviously; their peak levels were on the 7th day; the beta-gal level decreased on the 14th day; beta-gal expression disappeared in the most organs except the lungs on the 28th day. In all animals, the brain did not express any beta-gal. CONCLUSION: The adenovirus-mediated exogenous gene transfer in the internal jugular vein may be an effective approach of gene therapy in some diseases in the lung, liver, and kidney. PMID- 16710949 TI - Outbreak news. Cholera, Angola. PMID- 16710950 TI - Tetanus vaccine. PMID- 16710951 TI - Sedation in the intensive care unit: a call for evidence. PMID- 16710953 TI - Bullous lung disease or bullous emphysema? PMID- 16710952 TI - Recommendations of the 6th long-term oxygen therapy consensus conference. PMID- 16710954 TI - Novel recombinant norovirus in China. PMID- 16710955 TI - Rifampin-resistant Neisseria meningitidis. PMID- 16710956 TI - Vaccination-related Mycobacterium bovis BCG infection. PMID- 16710957 TI - Human Bocavirus in children. PMID- 16710958 TI - Extended-spectrum beta-lactamase-producing Enterobacteriaceae, Central African Republic. PMID- 16710959 TI - Novel recombinant sapovirus, Japan. PMID- 16710960 TI - Postmortem confirmation of human rabies source. PMID- 16710961 TI - Potential for zoonotic transmission of Brachyspira pilosicoli. PMID- 16710962 TI - Drug-resistant Mycobacterium tuberculosis, Taiwan. PMID- 16710963 TI - Enrofloxacin in poultry and human health. PMID- 16710965 TI - Philosophical and methodological foundations of psychiatric diagnosis. Introduction. PMID- 16710964 TI - Biodefense shield and avian influenza. PMID- 16710966 TI - Philosophical and methodological foundations of psychiatric diagnosis. Epilogue. PMID- 16710967 TI - Surgical management of acute epidural hematomas. AB - INDICATIONS FOR SURGERY: An epidural hematoma (EDH) greater than 30 cm3 should be surgically evacuated regardless of the patient's Glasgow Coma Scale (GCS) score. An EDH less than 30 cm3 and with less than a 15-mm thickness and with less than a 5-mm midline shift (MLS) in patients with a GCS score greater than 8 without focal deficit can be managed nonoperatively with serial computed tomographic (CT) scanning and close neurological observation in a neurosurgical center. TIMING: It is strongly recommended that patients with an acute EDH in coma (GCS score < 9) with anisocoria undergo surgical evacuation as soon as possible. METHODS: There are insufficient data to support one surgical treatment method. However, craniotomy provides a more complete evacuation of the hematoma. PMID- 16710968 TI - Surgical management of acute subdural hematomas. AB - INDICATIONS FOR SURGERY: An acute subdural hematoma (SDH) with a thickness greater than 10 mm or a midline shift greater than 5 mm on computed tomographic (CT) scan should be surgically evacuated, regardless of the patient's Glasgow Coma Scale (GCS) score. All patients with acute SDH in coma (GCS score less than 9) should undergo intracranial pressure (ICP) monitoring. A comatose patient (GCS score less than 9) with an SDH less than 10-mm thick and a midline shift less than 5 mm should undergo surgical evacuation of the lesion if the GCS score decreased between the time of injury and hospital admission by 2 or more points on the GCS and/or the patient presents with asymmetric or fixed and dilated pupils and/or the ICP exceeds 20 mm Hg. TIMING: In patients with acute SDH and indications for surgery, surgical evacuation should be performed as soon as possible. METHODS: If surgical evacuation of an acute SDH in a comatose patient (GCS < 9) is indicated, it should be performed using a craniotomy with or without bone flap removal and duraplasty. PMID- 16710969 TI - Website navigation and surgical education. AB - We evaluated the way in which medical students interacted with a surgical education Website. Ten medical students on their paediatric surgery rotation were asked to study online a chapter on intussusception in young children, for a period of 1 h. The computer was connected by video link to a video-recorder that recorded their navigation movements, i.e. navigation time, time spent in front of each page, stepwise, progressive exploration of the surgical contents, and use of interactive links. The students were asked to complete a questionnaire to identify their overall competency in computers and Internet navigation capabilities. The mean time spent viewing the entire Website was 48 min (range 31 63) and the mean time spent on the assigned chapter was 38 min (range 8-53). Each page out of 14 within the chapter was visited a mean of 1.8 times (range 0-10). Access to video selections occurred after a mean of 10 min after beginning the session. A connection was made to the video a mean of 2.3 times per session. The mean time spent viewing video footage was 9 min (24% of the time spent viewing the chapter). The questionnaire results showed that few students regularly accessed the Internet for educational purposes. PMID- 16710970 TI - Hospital consolidation: investigating causes and consequences. PMID- 16710971 TI - Prognosis for the uninsured: state lawmakers are still trying to do something about the millions of Americas [sic] without health care coverage, but the solution remains elusive. PMID- 16710972 TI - Power to the patient: can high-deductible plans and health savings accounts lower premium costs and replace the traditional goal of comprehensive insurance? PMID- 16710973 TI - [Relationship between the status of complementary alternative medicine and medical expenses on an isolated island in Okinawa--an analytical epidemiological study based on the details of medical expenses]. PMID- 16710974 TI - [Experimental study on CD40Ig gene transfer using adenovirus vector: a modified clamp technique in a rat model with liver transplantation]. PMID- 16710975 TI - [Relationship between medical expenses for the aged residing in farming villages and a social support network--documentation using details of medial expenses]. PMID- 16710976 TI - [Effect of L-carnitine to suppress the development of ischemic reperfusion injury in an excised heart of a diabetic rat]. PMID- 16710977 TI - [MLH1/MSH2 expression and its study as a prognostic factor in non-small cell lung cancer]. PMID- 16710978 TI - Human pythiosis. PMID- 16710979 TI - Rift Valley Fever potential, Arabian Peninsula. PMID- 16710980 TI - Screening and toxigenic corynebacteria spread. PMID- 16710981 TI - Rickettsia slovaca infection, France. PMID- 16710982 TI - Cutaneous anthrax, Belgian traveler. PMID- 16710983 TI - Japanese encephalitis, Singapore. PMID- 16710984 TI - HIV and lacaziosis, Brazil. PMID- 16710985 TI - Hand sanitizer alert. PMID- 16710986 TI - Spleen abscess as malaria complication. PMID- 16710987 TI - Rickettsioses in South Korea, materials and methods. PMID- 16710988 TI - Rickettsioses in South Korea, data analysis. PMID- 16710989 TI - "Mycobacterium tilburgii" infections. PMID- 16710990 TI - Access to health insurance: access to health insurance--2005. End of Year Issue Brief. PMID- 16710991 TI - [Application of navigation system for otorhinolaryngological surgery]. PMID- 16710992 TI - [Views on glaucoma of Czech ophthalmologists (results of the survey "What is your opinion?"]. PMID- 16710993 TI - [Recent progress in the diagnosis of otorhinolaryngological diseases--application of functional imaging of the brain]. PMID- 16710994 TI - Cloning and expression of apolipoprotein E3 and its variant apoE2 and apoE4. AB - In order to obtain three isoforms of apolipoprotein E (apoE), the cDNA encoding apoE3 was obtained by RT-PCR from normal human liver tissue. Site-directed mutagenesis was used to obtain the cDNAs encoding apoE2 and apoE4 isoforms. The 3 cDNAs were subcloned into vector pGEM-3Z and verified by DNA sequencing. The expression recombinant which can express the target protein as a (His) 6-tagged fusion was constructed by subcloning apoE cDNA into vector pT7-PL. The purified proteins were gained by Ni-NTA column. The SDS-PAGE results revealed the 6 His fusion proteins (apoE2, apoE3 and apoE4) were correctly expressed and purified successfully. PMID- 16710995 TI - Construction of shRNA targeted to the rat angiotensin II type 1 receptors and its RNAi in cytoplasma. AB - The expression vector of shRNA targeted to the rat angiotensin II receptor gene was constructed and the efficacy of siRNAs to modulate the expression of target gene in the in vitro cultured mammalian cells was investigated for antihypertensive therapy in spontaneous hypertensive rat (SHR) at post transcriptional level. The sense and antisense RNA oligonucleotides strands targeting angiotensin II receptor mRNA were synthesized individually according to the sequence of the rat angiotensin II receptor. For preparation of duplexes, sense- and antisense-stranded oligonucleotides were mixed and annealed, and the annealed duplexes were cloned into the pGenesil-1 vector. . The rat glioma cells were transfected with constructed pGenesil-1-shRNA plasmid and scrambled plasmid. The cultured cells were collected at different phases. RT-PCR and Western blot were performed. The AT1 mRNA and protein levels behaved ultimately same. Compared to control after 48 h, AT1 mRNA levels were decreased to 35.5% +/- 3.0%, and the levels reached their lowest point after 72 h (20.7% +/- 4% of control). At 24 and 48 h, AT1 protein was reduced to 46.9% +/- 4. 2% and 36.98% +/- 3.7% respectively compared to control and a maximum reduction was observed after 72 h of incubation (28.1% +/- 4% compared to controls). Plasmid-based shRNA expression systems targeted against the rat angiotensin II receptor gene were generated successfully. The shRNAs with a 22-nt stem and a short loop were cleaved into small interfering dsRNA (siRNA) by the Dicer. The in vitro transcribed siRNA enables the effective silencing of gene expression to the target mRNA and leads to effective inhibition of translation of proteins and will be lay the foundation of application of gene silencing technology to hypertensive rats. PMID- 16710996 TI - Construction of rat calcineurin A alpha cDNA recombinant adenovirus vector and its identification. AB - Rat calcineurin (CaN) A alpha isoform (Ppp3ca) cDNA recombinant adenovirus vector was constructed in order to explore the effect of CaN on the myocardium apoptosis induced by ischemia-reperfusion injury. Total RNA was isolated from the heart of the adult Wistar rat, and Ppp3ca CDS segment of approximate 1.59 kb size was amplified by reverse transcriptional PCR method. Ppp3ca cDNA segment was cloned into pMD18-T Simple vector for sequencing, and the right clone was named T Ppp3ca. Ppp3ca cDNA segment obtained from T-Ppp3ca was ligated with pShuttle2 IRES-EGFP to construct a recombinant plasmid pShuttle2-Ppp3ca-IRES-EGFP. Ppp3ca IRES-EGFP expression cassette containing CMV, Ppp3ca-IRES-EGFP and SV40 polyA DNA fragment (3.97 kb) obtained from pShuttle2-Ppp3ca-IRES-EGFP was connected with pAdeno-X backbone sequence to construct a recombinant plasmid pAdeno-Ppp3ca. After being identified by PCR and enzyme digestion, recombinant plasmid pAdeno Ppp3ca was packaged in HEK293 cells. Supernatant of adenovirus from HEK293 cells was collected after a visible cytopathic effect (CPE) appeared. The DNA of the recombinant adenovirus was extracted with the standard method. The presence of the recombinant adenovirus was verified by PCR. The results showed that sequencing results verified that the PCR product of Ppp3ca gene was identical to GenBank. Agarose electrophoresis showed the bands of recombined plasmid pAdeno Ppp3ca and the recombinant adenovirus identified by enzyme digestion and PCR were in the right range corresponding with expectation. It was concluded that the recombinant adenovirus carrying rat calcineurin A alpha (Ppp3ca) cDNA as well as a report gene-enhancer green fluorescent protein gene was successfully constructed in this experiment. PMID- 16710997 TI - Effect of hypoxic preconditioning on neural cell apoptosis and expression of Bcl 2 and Bax in cerebral ischemia-reperfusion in rats. AB - In order to investigate the protective effect of hypoxic preconditioning on the cerebral ischemia-reperfusion injury, the expression of Bcl-2 and Bax was detected by using immunohistochemical staining after 3 h cerebral ischemia followed by 1, 6, 12, 24 and 48 h reperfusion respectively in rats treated with or without hypoxic preconditioning before cerebral ischemia. In addition, the apoptosis of neural cells and the behavioral scores for neurological functions recovery were evaluated by TUNEL staining and "crawling method", respectively. Compared with control group (cerebral ischemia-reperfusion without hypoxic preconditioning), the expression of Bcl-2 was significantly increased, but that of Bax decreased in the hypoxic preconditioning group (cerebral ischemia reperfusion with hypoxic preconditioning), both P < 0.05. The pre-treatment with hypoxic preconditioning could reduce the apoptosis of neural cells and promote the neurological function recovery as compared to control group. It was suggested that hypoxic preconditioning may have protective effects on the cerebral ischemia reperfusion injury by inhibiting the apoptosis of neural cells, increase the expression of Bcl-2 and decrease the expression of Bax. PMID- 16710998 TI - Effects of ischemia and anoxia on cell activation and cell cycle of cultured astrocytes in vitro. AB - To examine the effects of ischemia and anoxia on cell activation and cell cycle of astrocytes in vitro, the cell cycles and the proliferation of astrocytes in different time points after ischemia and anoxia were studied by flow cytometry and BrdU labeling and the expression of GFAP and cyclin D1 was detected by the fluorescence immunochemistry. After ischemia and anoxia in vitro, the astrocytes in S phase were significantly increased as compared with those in the normal group and the proliferating ability of the astrocytes was highest 6 h after the treatment as revealed by BrdU pulse labeling, but the astrocytes in S phase and proliferating ability were decreased after 6 h. At the early stages of ischemia and anoxia, the positive staining intensity of GFAP was increased, peaked at 6th h, while 12 h after the ischemia and anoxia, the positive staining intensity of GFAP became weak, and the expression of cyclin D1 was gradually increased after the ischemic and anoxic damage. It is concluded that astrocytes are activated to proliferate and enter new cycle events by ischemia and anoxia, and cyclin D1 is implicated in the proliferation and repair of astrocytes. The cell cycle events are closely associated with the proliferation and activation of astrocytes. PMID- 16710999 TI - Effect of dexamethasone on expression of glucocorticoid receptor in human monocyte cell line THP-1. AB - The effect of dexamethasone with different concentrations and different stimulating periods on the expression of glucocorticoid receptors (GRalpha, GRbeta) protein was investigated in human monocyte cell line THP-1. The cultured human monocyte line THP-1 cells were stimulated by dexamethasone with different concentrations and different periods. The expression of GRalpha and GRbeta protein was detected by Western blotting. The results showed that the expression of GRalpha and GRbeta was detected in the THP-1 cells. The quantity of GRalpha expression was reduced by dexamethasone under the same concentration with the prolongation of the stimulating periods. The quantity of GRbeta expression was increased by dexamethasone treatment in a time- and dose-dependent manner. It was concluded that dexamethasone stimulation time-dependently reduced the GRalpha expression in THP-1 cells. Dexamethasone stimulation time- and dose-dependently increased the GRbeta expression in THP-1 cells. The expression of GRalpha and GRbeta was regulated by glucocorticoid. PMID- 16711000 TI - Correlation between endotoxin tolerance in human monocyte leukemia cell line THP 1 with glucocorticoid receptor-alpha. AB - Human monocyte leukemia cell line THP-1 was stimulated with lipopolysaccharide (LPS) to simulate the sepsis model and the expression of human glucocorticoid receptor-alpha (GR-alpha) mRNA in montocytes with endotoxin tolerance was investigated. THP-1 cells were cultured in serum-free medium, randomly divided into groups A, B, C, D and E, and stimulated with 0, 10, 10, 100, 0 ng/mL LPS for 24 h followed with 100, 100, 10, 100, 0 ng/mL LPS for another 24 h respectively. The expression of GR-alpha mRNA was detected by semi-quantitative reverse transcriptional polymerase chain reaction. Tumor necrosis factor-alpha (TNF alpha) was determined by enzyme linked immunosorbent assay (ELISA). The results showed that the A values of GR-alpha/beta-actin in groups A, B, C, D and E was 0.607 +/- 0.006, 0.368 +/- 0.005, 0.484 +/- 0.008, 0.509 +/- 0.004 and 0.564 +/- 0.014 respectively with the difference being significant among the groups (P < 0.05). The GR-alpha mRNA expression was negatively correlated with the TNF-alpha expression (P < 0.01). It was concluded that the down-regulation of the expression of GR-alpha mRNA in endotoxin tolerance THP-1 cells might play an important role in the development of endotoxin tolerance in THP-1 cells. PMID- 16711002 TI - Relationship between carbachol hyperstimulation-induced pancreatic acinar cellular injury and trypsinogen or NF-kappaB activation in rats in vitro. AB - The relationship between M3 cholinergic receptor agonist (carbachol) hyperstimulation-induced pancreatic acinar cellular injury and trypsinogen activation or NF-kappaB activation in rats was studied in vitro. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, the active protease inhibitor (pefabloc), and NF-kappaB inhibitor (PDTC) in vitro. Intracellular trypsin activity was measured by using a fluorogenic substrate. The cellular injury was evaluated by measuring the leakage of LDH from pancreatic acinar cells. The results showed that as compared with control group, 10(-3) mol/L carbachol induced a significant increase of the intracellular trypsin activity and the leakage of LDH from pancreatic acinar cells. Pretreatment with 2 mmol/L pefabloc could significantly decrease the activity of trypsin and the leakage of LDH from pancreatic acinar cells (P < 0.01) following the treatment with a high concentration of carbachol (10(-3) mol/L) in vitro. The addition of 10(-2) mol/L PDTC didn't result in a significant decrease in the activity of trypsin and the leakage of LDH from pancreatic acinar cells treated with a high concentration of carbachol (10(-3) mol/L) in vitro (P > 0.05). It was concluded that intracellular trypsinogen activation is likely involved in pancreatic acinar cellular injury induced by carbachol hyperstimulation in vitro. NF-kappaB activation may not be involved in pancreatic acinar cellular injury induced by carbachol hyperstimulation in vitro. PMID- 16711001 TI - Changes of expression of stretch-activated potassium channel TREK-1 mRNA and protein in hypertrophic myocardium. AB - The expression of stretch-activated potassium channel TREK-1 mRNA and protein of hypertrophic myocardium was measured. Using a model of hypertrophy induced by coarctation of abdominal aorta in male Wistar rats, the expression of TREK-1 mRNA and protein was detected by using semi-quantitative RT PCR and Western blot respectively. At 4th and 8th week after constriction of the abdominal aorta, rats developed significant left ventricular hypertrophy. As compared to sham-operated group, stretch-activated potassium channel TREK-1 mRNA was strongly expressed and protein was up-regulated in operation groups (P < 0.05). It was concluded that the expression of TREK-1 was up-regulated in hypertrophic myocardium induced by chronic pressure overload in Wistar rats. PMID- 16711003 TI - Influence of ciglitazone on A549 cells growth in vitro and in vivo and mechanism. AB - The effect and mechanism of the ciglitazone on lung cancer cells A549 growth in vitro and in vivo were studied. Various concentrations of ciglitazone were added to the cultured A549 line, and the proliferation and differentiation of A549 cells were examined by MTT and cytometry analysis. A549 cells (1 x 10(6)/mouse) were inoculated subcutaneously into 20 nude mice, which were randomly divided into two groups: the control group, the ciglitazone treated group. The weights of subcutaneous tumors were measured. The expression of cyclin D1 and P21 in the lung was detected by immohistochemistry and Western blot respectively. The results showed that the proliferation of A549 was inhibited significantly by ciglitazone in a dose- and time-dependent manner. There were more cells arrested in G1 /G0 phase and the expression of PPARgamma was markedly up-regulated in ciglitazone-treated group. Direct injection of ciglitazone into A549-induced tumors could suppress tumor growth in nude mice and the growth inhibitory rate was 36%. The expression of cyclin D1 was decreased and P21 increased significantly in ciglitazone-treated group as compared with control group. It was concluded that ciglitazone could inhibit A549 proliferation dose-dependently and time-dependently and induce differentiation, which might be related to the modulation of cell cycle interfered by PPARgamma. PMID- 16711004 TI - Influence of rmhTNF on the chemotherapy treatment of small cell lung cancer. AB - The efficacy and safety of the recombinant mutant human tumor necrosis factor (rmhTNF) combined with chemotherapy vs chemotherapy alone in the treatment of patients with small cell lung cancer (SCLC) were evaluated in this study. The selected 37 patients with SCLC were divided into experimental group (n =18) and control group (n = 19). Both groups were subjected to EP regimen. While in the experimental group, a regimen of 4 x 10(6) U/m2 rmhTNF intramuscular injection was given once a day from the 1st to 7th day and 11th to 17th day on the chemotherapy cycle. Twenty-one days were as a chemotherapy cycle and all patients received treatment with 2 cycles. The response rate was 83.3% (15/18) in the experimental group and 63.2% (12/19) in the control group respectively (P < 0.05). The KPS score after treatment was 78.4 +/- 9.6 in the experimental group and 71.2 +/- 9.7 in the control group with the difference being significant (P < 0.05). No severe adverse effects occurred in the two groups. It was concluded that the curative effectiveness of the rmhTNF combined with chemotherapy in the treatment of SCLC was more satisfactory than chemotherapy alone. The former could obviously improve the quality of life of the patients with SCLC. PMID- 16711005 TI - Mechanism of arsenic trioxide inhibiting angiogenesis in multiple myeloma. AB - In order to explore the molecular mechanism of arsenic trioxide treating multiple myeloma (MM) via inhibition of angiogenesis, the expression of brain derived neurotrophic factor (BDNF) and its specific receptor TrkB in human MM cell line KM3 and endothelial cell line ECV304 was detected by Western blotting. The angiogenic activity was evaluated by wound migration assay and tubule formation assay in vitro. The results showed that BDNF was detected in the MM cells and TrkB in the endothelial cells. Furthermore, 100 ng/mL BDNF could significantly induced endothelial cell tubule formation and wound migration. As2 O3 depressed the expression of BDNF and TrkB in the dose- and time-dependent manner. As2 O3 inhibited BDNF-induced wound migration and capillary tube formation. It was concluded that BDNF is a novel angiogenic protein as well as VEGF and has a relation with the pathogenesis of MM. As2 O3 interrupts a paracrine loop between MM cells and endothelial cells by down-regulating the TrkB expression in endothelial cells and inhibiting BDNF production in MM cells, finally resulting in inhibition of MM angiogenesis. This is probably one part of the mechanisms of the As2 O3 treating MM via the inhibition of angiogenesis. PMID- 16711006 TI - Allogeneic peripheral blood hematopoietic stem cell transplantation for patients with hematologic malignancies. AB - To investigate the therapeutic effects and associated complications of allogeneic peripheral blood stem cell transplantation (allo-PBSCT). 40 patients with various malignant hematopoietic diseases received allo-PBSCT. The preparative regimens were based on BUCY2 or modified BUCY2. The acute graft-versus host disease (aGVHD) was prevented by cyclosporin A and short-term MTX regimen in all patients. Two patients from donors with one fully mismatched HLA on DRB1 locus and 4 from unrelated donor also administered Zenapox (CD25 MAb) at dosage of 1 mg/kg every day on the day before transplantation and day 4 after transplantation. These 6 patients were also treated with mycophenolate mofetil (MMF). Transfusion of the donor cells: The median of the transfused nucleated cells was 5.38 x 10(8)/kg and that of the CD34+ cells was 7.8 x 10(6)/kg respectively. All the patients gained hematopoietic reconstruction except one who died of infection before engraftment. Seven patients got II degrees-IV degrees aGVHI) and the incidence was 17.5%. Fourteen patients got cGVHD and the incidence was 53.8% in the patients who survived over 6 months. Twenty-eight patients had fever or other characteristics of infection. The median follow-up time was 13.8 months. The incidence of transplantation related mortality (TRM) was 17.5% and 2 patients relapsed (5.0%). It was concluded that allo-PBSCT can reconstruct hematopoiesis quickly and is a favorable therapeutic method for leukemia. PMID- 16711007 TI - Influence of SB203580 on cell apoptosis and P38MAPK in renal ischemia/reperfusion injury. AB - The effects of SB203580 (SB) with different concentrations at different time points on renal function, apoptosis, P38MAPK activity and the expression, as well as the P38MAPK substrates in renal ischemia/reperfusion injury were investigated. Forty-nine rats were divided into 7 groups at random (n = 7 in each group) according to the durations of ischemia/reperfusion injury and the time of medication. Based on the orthogonal Latin side, the rats were injected, by caudal vein, with the same volume but different dosages of SB. BUN and Scr were determined. The apoptosis was detected with TUNEL kit. The protein was assayed qualitatively and semi-quantitatively by Western blot. The results showed that SB could significantly reduce the increased Scr and BUN, the apoptosis of renal tubular epithelia and the activation of P38MAPK all caused by renal ischemia/ reperfusion injury in a dose-dependent manner (P < 0.05). And the effect was most predominant when SB was given 3 h before renal ischemia. This suggested that SB could significantly alleviate renal ischemia/reperfusion injury. Administration of SB 3 h before ischemia at the concentration of 5 micromol/L could obtain an optimal effect. PMID- 16711008 TI - Effects of 17beta-estradiol on the expression of IL-6, IL-11 and NF-kappaB in human MG-63 osteoblast-like cell line. AB - In order to characterize the effects of 17beta-estradiol (17beta-E2) on the expression of IL-6, IL-11 and NF-kappaB in the human MG-63 osteoblast-like cell line, the expression of IL-6 was detected by RT-PCR, Northern blot and Western blot. The expression of IL-11 was determined by RT-PCR, and NF-kappaB by Western blot. The results showed that 17beta-E2 down-regulated the expression of IL-6 mRNA and protein, IL-11 mRNA and NF-kappaB protein in MG-63 cells. It was suggested that the expression of NF-kappaB, IL-6 and IL-11 in MG-63 cells could be suppressed by 17beta-E2, and this might lend support to estrogen replacement therapy in postmenopausal women. PMID- 16711009 TI - Perihematoma damage at different time points in experimental intracerebral hemorrhage. AB - The damage degree of neurons in perilesion at different time points was observed in order to explore the optimal operation occasion. Piglet lobar hematomas were produced by pressure-controlled infusions of 2.5 mL autonomous blood into the right frontal hemispheric white matter over 15 min, and the metabolic changes were ambulatorily detected with MRS at 3rd, 12th, 24th and 48th h after hematoma induction. Brain tissues of perihematoma were also obtained at different time points. The transcription level of Bax gene was detected by in situ hybridization and apoptosis by TUNEL technique, and the pathologic change of neurons was observed under an electron microscope. The results showed that the number of Bax positive cells reached the peak at 24 h (79.00 +/- 4.243/5 fields). There was no significant difference in A values between 3 h and 6 h, 12 h (P > 0.05), but there significant difference between 24 h and 3 h, 6 h, 12 h (P < 0.05). The number of apoptotic cells reached the peak at 24 h (P < 0.001), and there was no significant difference between 3 h and 6 h (P = 0.999). The area of the apoptotic cells showed no significant difference between 3 h and 6 h or among 3 h, 6 h and 6 h (P > 0.05). Lac peak mainly occurred at 24 h and 48 h, while on the healthy side, no Lac peak was detectable. The ratio of NAA/Cr presented a descent tendency, but there was no significant difference among the groups before 12 h (P > 0.05), there was very significant difference between 3, 6 and 24, 48 h (P < 0.01). Under electronic microscopy, the neuronal damage surrounding hematoma in 3 to 6 h was milder than in 24 h to 48 h. It was concluded that the secondary apoptosis, damage and metabolic disturbance of the neurons surrounding hematoma was milder in 3-6 h in acute intracerebral hemorrhage, while obviously aggravated in 24-48 h. An effective intervention is needed to reduce secondary damage as soon as possible. PMID- 16711010 TI - A possible association of Staphylococcus enterotoxin B-induced asthma and sinusitis. AB - In order to gain insight into a possible association between chronic sinusitis and asthma, 85 patients with sinusitis and asthma underwent functional endoscopic sinus surgical treatment and serum antibodies and cytokines were measured. The results showed that 51 out of 85 patients with high serum anti Staphylococcus enterotoxin B (SEB) antibody before treatment obtained satisfactory results for both sinusitis and asthma. The high level of Th2 cytokine IL-4 was down regulated to the levels of normal controls after sinus surgery. Thirty-four out of 85 patients did not show high serum anti-SEB antibody before sinus surgery and did not show much improvement in their asthmatic symptoms although sinusitis symptoms were resolved by sinus surgery. It was concluded that bacterial superantigen SEB (in the sinuses) might play a crucial role in the pathogenesis of lower airway hypersensitivity. PMID- 16711011 TI - Inhibition of metastatic progression of SSTR2 gene transfection mediated by adenovirus in human pancreatic carcinoma cells. AB - The inhibition of metastatic progression of Somatostatin receptor type 2 (SSTR2) gene transfection mediated by adenovirus in human pancreatic carcinoma cells and the mechanisms involved in this effect were studied. The full-length human SSTR2 cDNA was introduced into the pancreatic cancer cell line BXPC-3 by adenovirus mediated transfection. Stable expression of mRNAs and protein of SSTR2 was detected by RT-PCR and Western-blot. The Matrigel-coated Transwell was used to detect the migratory and invasive ability of SSTR2-expressing cells, Adv-GFP control cells and mock control cells. Furthermore, the expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) was detected by RT-PCR in these cells. The stable expression of SSTR2 was detected in BXPC-3 transfected by Adv-GFP-SSTR2. A dramatic decrease of BXPC-3 expressing sst2 cells migrating through a Matrigel-coated filter was observed, as compared with Adv-GFP control and mock control cells (P < 0.01). Moreover, the expression of MMP-2 mRNA was significantly reduced in the SSTR2-expressing cells and conversely the expression of TIMP-2 mRNA was significantly increased in the SSTR2-expressing cells when compared with the Adv-GFP control and mock control (P < 0.01). The expression of reintroduced human SSTR2 gene in BXPC-3 cells by Adv GFP-SSTR2 had the anti-migratory and anti-invasive effects, and the mechanisms involved in this effect may be due to the down-regulated expression of MMP-2 and up-regulated expression of TIMP-2. PMID- 16711012 TI - Transfection of antisense oligodeoxynucleotide inhibits heparanase gene expression and invasive ability of human pancreatic cancer cell in vitro. AB - Extracellular matrix (ECM) degradation is an essential step that allows tumor cells to penetrate a tissue barrier and become metastatic. Heparanase (HPSE) is an endoglycosidase that specifically degrades heparin sulfate proteoglycans (HSPG), a chief component of ECM. HPSE is not expressed in normal epithelial cells but can be detected in a variety of human carcinomas including pancreatic cancer. In the present study, human pancreatic cancer cell line Panc-1 was transfected with HPSE antisense oligodeoxynucleotide (AS-ODN) in vitro, then the inhibitory effect of AS-ODN on HPSE gene expression and invasive ability of Panc 1 cells in vitro was examined. The HPSE mRNA and protein expression of Panc-1 cells transfected with AS-ODN was significantly inhibited. However, there were no marked inhibitory effects in Panc-1 cells treated with nonsense oligodeoxynucleotide (NS-ODN). Moreover, a modified Boyden chamber assay demonstrated that transfection with HPSE AS-ODN significantly inhibited invasive potential of Panc-1 cells in vitro after AS-ODN transfection. This suggests that HPSE AS-ODN may contribute to the inhibition of HPSE mRNA and protein expression, and results in a decrease of the invasive ability of Panc-1 in vitro. PMID- 16711013 TI - FasL expression in colorectal carcinoma and its significance in immune escape of cancer. AB - To study the significance of FasL expression in immune escape of colorectal carcinoma, FasL protein expression and the number of tumor infiltrating lymphocytes (TILs) in 80 specimens of colorectal carcinoma were detected by immunohistochemitry. The mRNA of FasL was measured by in situ hybridization in the consecutive tissue slices of 80 colorectal carcinomas respectively. Using terminal deoxynucleotidyl transferase-mediaed dUTP nick end labeling (TUNEL), apoptotic cells were detected in 80 specimens of colorectal carcinoma. The expression of FasL was detected in all 80 specimens, but it was not even in the same or among different tissues. In the consecutive tissue slices, the location of expression of FasL protein corresponded with that of FasL mRNA. In those with FasL extensive expression, the number of TILs was less than that of FasL weak expression (P < 0.05), and the apoptotic index (AI) of TILs was higher and that of tumor cells was lower than that of FasL with weak expression respectively (P < 0.01). The Al of TILs was correlated with that of tumor cells (r = -0.631, P < 0.01). It was suggested that colorectal carcinoma cells can induce the apoptosis of TILs through the expression of FasL, which can counterattack the immune system. This may be one of the mechanisms of immune evasion in colorectal carcinoma. PMID- 16711014 TI - Total arterial revascularisation in left ventricular dysfunction. AB - The feasibility and safety of total arterial coronary revascularization with 2 arterial conduits in patients with impaired left ventricular function was evaluated. Data were prospectively collected on all patients with multiple vessel discase and moderately or severely impaired left ventricular function, who underwent coronary surgery with the intention of total arterial revascularization with 2 conduits between March 1995 and August 2002. One hundred and seventy-nine patients were included in the study. Acute coronary insufficiency was present in 3 patients and 43 had unstable angina. Severe left ventricular impairment was present in 29 patients. There were 17 redo operations including 3 redo-redo procedures. Eighty-two percent of patients had a Y graft configuration from the left internal mammary artery (right internal mammary artery 40.8%, radial artery 33.5%, other 7.8%). The perioperative mortality was 2.2%, myocardial infarction 1.7% and stroke 0.6%. Total arterial revascularization in patients with ischaemic left ventricular dysfunction can be safely performed with 2 arterial conduits. The radial artery provides conduit length greater than the right internal mammary artery and allows full revascularization despite left ventricular dilatation. PMID- 16711015 TI - Expression of EGFR and LRIG-1 in human trigeminal neurinoma. AB - The expression of epidermal growth factor receptor (EGFR) and leucine-rich repeats and immunoglobulin-like domain 1 (LRIG-1) in human trigeminal neurinoma was investigated and their effect on the origination and development of trigeminal neurinoma, and the relationship between them was studied. By using immunohistochemistry with tissue chip, the expression of EGFR and LRIG-1 was detected in 23 cases of trigeminal neurinoma. It was found that in the 23 cases, the expression rate of EGFR was 21.74%, while that of the LRIG-1 was 78.26%. There was a negative correlation between them. It was suggested that LRIG-1 might inhibit the malignant differentiation and proliferation of the trigeminal neurinoma possibly by the negative feedback loop of EGFR. PMID- 16711016 TI - Treatment and prognosis of pituitary adenomas in children. AB - In order to explore the treatments and prognosis of pituitary adenomas in children, the experience in the treatment of 15 children with pituitary adenomas was summed up. The clinical data of 15 children with pituitary adenomas in Tongji Hospital were retrospectively analyzed from Jan. 1997 to Aug. 2004, including 9 males and 6 females with the age ranging from 9 - 15 years old (mean 12.8). The disease duration was 3 months to 5 years (mean 2 years). Fourteen cases were subjected to the microsurgical operation through oral tanssphenoidal approach and one case to medical treatment. All cases were followed up from 1 to 5 years by imaging studies and endocrine investigation. The results showed that 13 cases obtained total removal of the tumor and one subtotal removal of tumor. No severe complications and operation death occurred. It was concluded that the tanssphenoidal approach was the optimal treatment for children with pituitary adenomas. Regular and long-term follow-up is of great importance. PMID- 16711017 TI - Treatment of floating knee injury in children. AB - The necessity and superiority of the surgical operation on children with floating knee injury and the fracture union and complications were investigated. Twenty eight children with floating knee injury were subjected to open reduction and internal fixation or external fixator. The patients were followed up for 18 months to 7 years. The curative effectiveness was scored by Karlstrom criteria. The results showed that no nonunion or deformity was found. The affected limb was 1.2 cm to 1.5 cm longer in 2 cases, 0.8 to 1.2 cm shorter in 3 cases than the contralateral. No severe dysfunction of knee joint occurred. The excellent-good rate was 92.8% and the curative rate 71.4% respectively. So for children whose age is older than 5 years, it's a good way to treat the fractures of femur and tibia with open reduction and internal fixation or external fixator. The method can be advantageous for the nursing care, early function recovery, shortening of the hospital stay and avoidance of severe complications. PMID- 16711018 TI - Improvements of surgical technique in establishment of rat orthotopic pulmonary transplantation model using cuffs. AB - In order to establish more simple and effective rat orthotopic lung transplantation models, 20 rats were divided into donor and recipient groups. Rat lung transplantation models were established by using improved cuff technique. All the 10 operations were accomplished successfully. The mean operative time of recipients was 45 +/- 4 min. The survival time was over 30 days after lung transplantation. The checks of X-ray were almost normal. There was no significant difference in the blood gas analysis before and after clipping the right hilum (P > .05). This method is more simple, applicable and requires less time. PMID- 16711019 TI - Promoter hypermethylation of DNA repair gene MGMT in laryngeal squamous cell carcinoma. AB - The relationship between hypermethylation of CpG islands in the promoter regions of O6-methylguanine DNA methyltransferase (MGMT) genes and laryngeal squamous cell carcinoma was explored. Methylation-specific PCR and semi-quantitative RT PCR were used to study the promoter methylation and mRNA expression of the MGMT gene in laryngeal carcinoma tissues, tissues adjacent to the tumor and normal laryngeal tissues. Hypermethylation of MGMT gene was detected in 16 samples of 46 (34.8%) laryngeal squamous cell carcinoma samples. However, the MGMT hypermethylation was not detected in all tissues adjacent to the tumors and normal tissues. No significant difference in MGMT gene hypermethylation was found in samples with different histological grades (chi2 = 3.130, P = 0.077) or in samples from patients with different TNM status (chi2 = 3.957, P = 0.138). No expression of MGMT mRNA was detected in all hypermethylated laryngeal carcinoma tissues. The expression of MGMT mRNA was detected in all unmethylated laryngeal carcinoma tissues, tissues adjacent to the tumors and normal tissues. It suggests that MGMT gene promoter hypermethylation is associated with MGMT gene transcription loss in laryngeal carcinoma tissues and possibly plays an important role in carcinogenesis of laryngeal tissues. PMID- 16711020 TI - Expression of vascular endothelial growth factor and cyclooxygenase-2 in laryngeal squamous cell carcinoma and its significance. AB - In order to study the expressions of vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2) in human laryngeal squamous cell carcinoma (LSCC) and its significance, the expression of VEGF mRNA and COX-2 mRNA in 62 cases of LSCC and 54 adjacent noncancerous laryngeal tissues and 9 normal human laryngeal mucous tissues was detected by using techniques of semi-quantitative RT-PCR. It was found that the expression level of VEGF and COX-2 mRNA was significantly increased in LSCC as compared with that in the normal human laryngeal mucous tissues (both P < 0.01), and the expression level of VEGF and COX-2 mRNA were significantly increased in stage Ill + IV tissues of LSCC as compared with the stage I + II tissues of LSCC (P < 0.01). There was a high positive correlation between VEGF and COX-2 expression in LSCC (r = 0.756, P < 0.01). These data raise the possibility that VEGF and COX-2 may play key roles in the growth, invasion and metastasis of LSCC. PMID- 16711021 TI - Partial horizontal laryngectomy and epiglottiplasty. AB - In order to evaluate the availability of the lateral horizontal laryngectomy and anaplasty of epiglottis to treat some patients with specific supraglottic carcinomas and hypopharyngeal carcinomas, 17 cases of laryngeal and hypopharyngeal carcinomas were retrospectively analyzed, whose tumors were located at the lateral margin of epiglottis, aryepiglottic fold, medial wall of piriform fossa and were treated by the lateral horizontal laryngectomy and anaplasty of epiglottis. The results showed that all cases took food by mouth in postoperative 9-14 days and subjected to decannulation in postoperative 9-15 days. Three cases had postoperative hoarse voice. The free-disease survival rate of 3 years was 71.4% in 14 cases followed up after the first surgical therapy, and the overall free-disease survival rate of 3 years was 85.7% after the second surgical therapy. It was concluded that the manipulations of the lateral horizontal laryngectomy and epiglottiplasty were simple. It could alleviate the postoperative symptoms of aspiration and bucking remarkably and shorten their postoperative recovery time, yet does not lower the survival rate of patients if laryngocarcinoma or hypopharyngeal carcinoma cases were properly selected. PMID- 16711022 TI - Effects of antisense oligodeoxynucleotide to follicle-stimulating hormone receptor on the expression of proliferating cell nuclear antigen and vascular endothelial growth factor in primary culture cells derived from human ovarian mucinous cystadenocarcinoma. AB - The effects of antisense oligodeoxynucleotide (antisense ODN) to follicle stimulating hormone receptor (FSHR) and follicle-stimulating hormone (FSH) on the expression of proliferating cell nuclear antigen (PCNA) and vascular endothelial growth factor (VEGF) were studied in primary culture cells derived from human ovarian mucinous cystadenocarcinoma (OMC). The primary OMC cells were cultured with the enzyme digestion method, and the expression of pan Keratin protein and FSHR mRNA was detected for identification of the cells. OMC cells were co cultured with antisense ODN, nonsense ODN and FSH with different concentrations for 48 h and 72 h. The expression of PCNA and VEGF was detected by using SP immunohistochemistry. Compared with that in the control group, the PCNA and VEGF expression was increased obviously in FSH groups (P < 0.05 or P < 0.01), while decreased significantly in antisense ODN groups (P < 0.05 or P < 0.01) and unchanged in nonsense ODN groups, respectively. Meanwhile, antisense ODN could antagonize the increased expression of PCNA and VEGF caused by FSH significantly (P < 0.01). It was suggested that FSH might promote the development of OMC to some extent. Antisense ODN could inhibit the proliferative activity of OMC cells and the promoting proliferative activity enhanced by FSH. PMID- 16711023 TI - Analysis of placental growth factor in placentas of normal pregnant women and women with hypertensive disorders of pregnancy. AB - To investigate the expressions of placental growth factor (PLGF) in placenta with hypertensive disorders of pregnancy (HDP), 45 women with HDP and 20 normally pregnant women were studied. Among 45 women with HDP, there were 23 cases of severe preeclampsia and one case of eclampsia. The location and level of PLGF proteins was determined by immunohistochemistry and Western blot. The expression of PLGF mRNA in placenta was assessed by reverse transcriptional-polymerase chain reaction (RT-PCR). The results showed that: (1) The distribution of PLGF in placenta with HDP was similar to normal one, which was mainly in the cytoplasm of villous syncytiotrophoblast and villous stroma; (2) The expression of PLGF protein was significantly decreased in placentas with mild and severe preeclampsia compared to the normal ones (0.3 +/- 0.4 vs 0.6 +/- 0.4, 0.2 +/- 0.5 vs 0.6 +/- 0.4, P < 0.01). There were no differences between the gestational hypertension placenta and normal one (0.5 +/- 0.6 vs 0.6 +/- 0.4, P > 0.05); (3) The transcription levels of the PLGF mRNA in placentas with preeclampsia were significantly lower than in normal groups (3.33 +/- 0.39 vs 4.87 +/- 0.60, 1.97 +/- 0.29 vs 4.87 +/- 0.60, P < 0.01), and no differences were found between the gestational hypertension placenta and normal groups. These findings suggest that the abnormal expression of PLGF in placentas is related to the pathogenesis of HDP. PMID- 16711024 TI - Expression of Pin1 and Ki67 in cervical cancer and their significance. AB - In order to investigate the expression levels of Pin1 mRNA and protein in cervical cancer and its association with Ki67 and their clinical significance, amplification of Pin1 gene was examined by RT-PCR, and the expression of both Pin1 and Ki67 protein was detected by immunohistochemistry in cervical cancer tissues. It was shown that the expression levels of Pin1 were higher in cervical cancer than in normal cervical tissues (P < 0.05). The expression of Pin1 protein was increased progressively along with the disease process from normal cervix to CIN and to cervical cancer (P < 0.05). No significant difference in the Pin1 expression was found between disease stages (FIGO), pathological grades or pelvic lymph node metastasis status (P > 0.05). The expression of Pin1 was significantly higher in adenocarcinoma than in squamous carcinoma of the uterine cervix (P < 0.05). In cervical cancer, the overexpression of Pin1 was positively correlated with that of Ki67 (P < 0.05). These results suggested that the overexpression of Pin1 was closely related with cancer cell proliferation or progression of cervical cancer and contributed to oncogenesis. Pin1 may serve as a potential marker for cervical cancer diagnosis. PMID- 16711025 TI - Detection of human parvovirus B19 nonstrutural protein DNA by nested-polymerase chain reaction in gravida serum and pregnant tissues. AB - A new nested-polymerase chain reaction (nested-PCR) assay was developed to detect human parvovirus B19 DNA corresponding to the nonstructural protein in clinical specimens in a routine diagnostic laboratory. The sensitivity of this highly specific assay was up to 0.005 fg of B19 DNA. Parvovirus B19 was identified in sera of 20 pregnant women with abnormal pregnant outcome. Among these 20 cases, intrauterine parvovirus infection did exist in 7 pregnant women because parvovirus B19 DNA was detected in the pregnant tissues of them such as placenta tissues, chorionic villi, amniotic fluid, fetal spleen, liver and abdominal fluids. PMID- 16711026 TI - Effect of semen quality on the embryo development. AB - To investigate the influences of sperm quality on the zygotes and embryos development, as the role of the paternal factor in early human embryogenesis is gaining more attention because of the application of techniques such as intracytoplasmic sperm injection (ICSI) for the treatment of men infertility. 136 infertility couples with men factors (Group I ) were included from May 2002 to January 2001. One hundred and seventy-two infertility couples with tube factors (Group II) served as controls. The sperm parameters, gemmates and embryos quality, implantation rate and pregnant rate in both groups were analyzed. It was found that there was no significant differences in the number of oocytes retrieved, the fertilization rate and number of embryos transferred between two groups. Sperm concentration, percentage of motile sperm and percentage of sperm with normal morphology were significantly lower in group I than in group II (P < 0.01). The proportion of good quality zygotes and good quality embryos were significantly lower in the male infertility group than in the tubal disease group (P < 0.05). Implantation rate and pregnancy rate were similar in two groups. It was concluded that spermatozoa is involved in the embryo quality, even in the early stages of development, which limited the treatment potency of IVF procedure. PMID- 16711027 TI - Effects of murine cytomegalovirus infection on sperm viability in mice. AB - In order to explore the effects of testicular infection of murine cytomegalovirus (MCMV) on mature sperm viability at different periods following MCMV inoculation in mice, 91 BALB/c mice without MCMV infection were randomly divided into two groups: an experimental group (n = 56) and a control group (n = 35). The mice in the experimental group were treated by inoculating MCMV intratesticularly, while those in the controlled group were directly inoculated with DMEM without MCMV. The mice in both groups were sacrificed separately on the day 1, 1. 5, 2, 4, 6, 9 and 14 post-inoculation (D1) 1. 5, 2, 4, 6, 9 and 14 PI). The MCMV M83 mRNA gene was detected in the testis by in situ hybridization (ISH) with MCMV late-mRNA probe labeled with digoxin. Sperm viability of mature sperm in the epididymis cauda was measured. The results demonstrated the positive signal of ISH of MCMV was found mainly in the cytoplasm of the testicular interstitial cells and spermatogenic cells in the experimental group. Compared with that in the controlled group, the sperm viability in the experimental group was decreased significantly on D1 PI and D1.5 PI (P < 0.05). No statistically significant difference in the sperm viability was found after D2 PI between two groups (P > 0.05). This suggested that sperm viability in mice might be descended significantly shortly after MCMV infection and might return to normal with time, indicating that MCMV acute infection might temporarily degrade sperm quality and influence procreation transiently. PMID- 16711028 TI - Effect of shRNA inhibiting HiF1alpha gene on TIMP1 expression in RPE cells. AB - Small hairpin RNA (shRNA) was used to silence the HIF1alpha gene in human retinal pigment epithelial cells (RPE) under hypoxia in order to observe the effect of gene silencing on the expression of matrix metalloproteinase tissue inhibitor 1 (TIMP1). By using chemical hypoxic inducer CoCl2, to mimic RPE hypoxic environment, shRNA against the targeting region of HIF1alpha mRNA sequence was synthesized by a method of in vitro transcription, and the HIF1alpha was interfered in RPE cultured under hypoxia (induced by 150 micromol/L CoCl2). RT PCR was employed to detect the expression of HIF1alpha and TIMP1. The expression levels of HIF1alpha and TIMP1 were measured by using Western blotting. The results showed that after the RPE were transfected with specific shRNA against HIF1alpha mRNA, RT-PCR revealed that under hypoxia, the efficacy of HIF1alpha gene silencing in RPE was 83.4%. Western blotting revealed that the expression levels of HIF1alpha protein was dramatically dropped. In addition. RT-PCR results demonstrated that the expression of TIMP1 mRNA was decreased by 28.9%, and the expression levels of TIMP1 protein were also significantly reduced by Western blotting. It was suggested that shRNA targeted against HIF1alpha mRNA could effectively silence the HIF1alpha gene, subsequently effectively inhibit the hypoxia-induced up-regulation of TIMP1. PMID- 16711030 TI - Central corneal thickness in adult Chinese. AB - The central corneal thickness (CCT) in age 48 years or less of Chinese was characterized and its relationship with gender, age, refraction and intraocular pressure (IOP) was investigated. Right eyes of 1669 participants were included in this study (880 men, 52.7% and 789 women, 47.3%). Mean age of the samples was 23.8 +/- 5.9 years. After the examination of corneal topography and refraction, Goldman applanation tonometry was carried out by one physician. Tonometric values were the mean of three consecutive readings. Subsequently, another physician carried out ultrasonic pachymetry with the DGH 2000 AP ultrasonic pachymeter. Six measurements were made at the center of the cornea of each eye. The mean value was used for analysis. The results showed that mean CCT of male participants was 551.33 +/- 34.62 microm, 5.79 microm more than that of female participants. Linear regression analyses revealed that CCT was negatively related with age only in female and no association was found between refractive status and CCT. IOP was positively related to CCT, and there was a difference in IOP of 1.5 mmHg (1 mmHg = 0.133 kPa) per 100 microm difference in CCT. Ocular hypertension group was prone to have thicker cornea than average. The results indicated that in adult Chinese CCT tended to decrease with aging in female only. IOP measured by Goldmann tonometry was positively related with CCT so that CCT should be measured along with IOP. PMID- 16711029 TI - Effect of dexamethasone and aquaporin-1 antisense oligonucleotides on the aquaporin-1 expression in cultured human trabecular meshwork cells. AB - The changes in the expression of aquaporin-1 (AQP1) mRNA and protein in cultured human trabecular meshwork (HTM) cells treated with dexamethasone and transfected with antisense oligonucleotides (AS-ODN) were studied, and the implication of AQP1 regulation in corticosteroid-glaucoma and the possibility of AS-ODN inhibiting the AQP1 expression were evaluated. The cultured HTM cells in vitro were treated with different concentrations of dexamethasone and transfected with oligonucleotides for 5 days respectively. Then, total RNA and protein of HTM cells were extracted. The changes of AQP1 mRNA and protein were demonstrated qualitatively and quantitatively by RT-PCR and Western blot. Band intensities were detected by imaging analysis. There was a parallel relationship between the results of RT-PCR and those of Western blot. The expression levels of AQP1 mRNA and protein in dexamethasone-treated groups were increased initially and decreased later as dexamethasone concentration was stepped up. In the 0. 04 microg/mL and 0.4 microg/mL groups, the levels of AQP1 were higher than in control group (0 microg/mL). In the 4 microg/ mL and 40 microg/mL groups, the AQP1 expression levels were lower than in control group. AS-ODN could down regulate the expression of AQP1 mRNA and protein in a dose-dependent manner. At 5 microg/mL, down-regulation efficiency reached the maximum. There was no statistically significant difference in the expression of AQP1 mRNA and protein between all sense oligonucleotides groups and control group. It was suggested that dexamethasone may induce the changes of the AQP1 expression in HTM cells to be involved in the occurrence of corticosteroid-glaucoma. AS-ODN can down regulate the AQP1 expression in HTM cells to some extent. PMID- 16711031 TI - Tc1/Tc2 imbalance in the peripheral blood of patients with recurrent genital herpes. AB - In order to investigate the IFN-gamma and IL-4 expression of CD8+ T lymphocytes in the peripheral blood from patients with recurrent genital herpes (RGH) at different clinical periods and their relationship with the pathogenesis of RGH, flow cytometry was used to detect the intracellular cytokines (IFN-gamma and IL 4) of CD8+ T lymphocytes in the peripheral blood of 30 patients with RGH at acute period, 20 patients with RGH at recovery period and 15 healthy volunteers. The results showed that RGH patients at acute period had a lower percentage of Tc1 subsets in peripheral blood than that of healthy controls (P < 0.001), especially a remarkable decreased percentage of Tc1 subsets (P < 0.001) among those RGH patients with recurrent number more than 3 in the recent half a year. Tc1/Tc2 ratio in the RGH patients at acute period was significantly decreased as compared with normal control group (P < 0.05). The recurrent number of acute patients in the recent half a year was significantly correlated with the percentage of Tc1 subsets and the ratio of Tc1/Tc2 (P < 0.05). A decreased percentage of Tc1 subsets was found among the RGH patients with recurrent number more than 3 in the recent half a year at recovery period in comparison with healthy volunteers (P < 0.05), and it was significantly correlated with the recurrent number in the recent half a year (P < 0.05). It is concluded that there are Tc1/Tc2 imbalance and a low level of Tc1 subsets in RGH patients who are relapsing repeatedly in the near period. The low level of Tc1 subsets may be an important factor for the recurrence of RGH and the reactivation of latent herpesvirus infection. PMID- 16711032 TI - Real-time three-dimensional color Doppler flow imaging: an improved technique for quantitative analysis of aortic regurgitation. AB - The recently introduced real-time three-dimensional color Doppler flow imaging (RT-3D CDFI) technique provides a quick and accurate calculation of regurgitant jet volume (RJV) and fraction. In order to evaluate RT-3D CDFI in the noninvasive assessment of aortic RJV and regurgitant jet fraction (RJF) in patients with isolated aortic regurgitation, real-time three-dimensional echocardiographic studies were performed on 23 patients with isolated aortic regurgitation to obtain LV end-diastolic volumes (LVEDV), end-systolic volumes (LVESV) and RJV, and then RJF could be calculated. The regurgitant volume (RV) and regurgitant fraction (RF) calculated by two-dimensional pulsed Doppler (2D-PD) method served as reference values. The results showed that aortic RJV measured by the RT-3D CDFI method showed a good correlation with the 2D-PD measurements (r = 0.93, Y = 0.89X + 3.9, SEE = 8.6 mL, P < 0.001); the mean (SD) difference between the two methods was--1.5 (9.8) mL. % RJF estimated by the RT-3D CDFI method was also correlated well with the values obtained by the 2D-PD method (r = 0.88, Y = 0.71X + 14.8, SEE = 6.4%, P < 0.001); the mean (SD) difference between the two methods was--1.2 (7.9) %. It was suggested that the newly developed RT-3D CDFI technique was feasible in the majority of patients. In patients with eccentric aortic regurgitation, this new modality provides additional information to that obtained from the two-dimensional examination, which overcomes the inherent limitations of two-dimensional echocardiography by depicting the full extent of the jet trajectory. In addition, the RT-3D CDFI method is quick and accurate in calculating RJV and RJF. PMID- 16711033 TI - Left ventricular regional systolic function in patient with hypertrophic cardiomyopathy by quantitative tissue velocity imaging. AB - The left ventricular regional systolic functions in patients with hypertrophic cardiomyopathy (HCM) were assessed by using quantitative tissue velocity imaging (QTVI). Left ventricular (LV) regional myocardial velocity along long- and short axis in 31 HCM patients and 20 healthy subjects were analyzed by QTVI, and the regional myocardial systolic peak velocities (MVS) were measured. Mean MVS at each level including mitral annular, basal, middle and apical segments were calculated. The ratio of MVS along long-axis to that along short-axis (Ri) at basal and middle segments of the LV posterior wall and ventricular septum were calculated. The results showed that mean MVS was slower at each level including mitral annular, basal, middle and apical segments in the HCM patients than that in the healthy subjects (P < 0.01). There were no significant differences in mean MVS between obstructive and non-obstructive groups in HCM patients. MVS of all regional myocardial segments along long-axis in the HCM patients were significantly slower than that in the healthy subjects (P < 0.05), but there was no significant difference in MVS of all regional myocardial segments along long axis between hypertrophied and non-hypertrophied group in the HCM patients. Ri was significantly lower in the HCM patients than that in the healthy subjects. The LV regional myocardial contractility along long-axis was impaired not only in the hypertrophied wall but also in the non-hypertrophied one in patients with HCM, suggesting that QTVI can assess accurately LV regional systolic function in patient with HCM and provides a novel means for an early diagnosis before and independent of hypertrophy. PMID- 16711034 TI - Hindcasting nitrogen deposition to determine an ecological critical load. AB - Using an estimated background nitrogen (N) deposition value of 0.5 kg N x ha(-1) x yr(-1) in 1900, and a 19-year record of measured values from Loch Vale (Colorado, USA; NADP site CO98), I reconstructed an N-deposition history using exponential equations that correlated well with EPA-reported NO(x) emissions from Colorado and from the sum of emissions of 11 western states. The mean wet N deposition values for the period 1950-1964 was approximately 1.5 kg N x ha(-1) x yr(-1), corresponding to the reported time of alteration of diatom assemblages attributed to N deposition in alpine lakes in Rocky Mountain National Park (USA). This value becomes the critical load defining the threshold for ecological change from eutrophication. Thus if an N-deposition threshold for ecological change can be identified, and the date at which that threshold was crossed is known, hindcasting can derive the amount of atmospheric deposition at the time of change, at least for alpine lakes. Independent support for the technique and the deposition amount comes from experimental studies, ecosystem modeling, and paleolimnological records from northern Wyoming (USA). PMID- 16711035 TI - The value of small size: loss of forest patches and ecological thresholds in southern Madagascar. AB - Many services generated by forest ecosystems provide essential support for human well-being. However, the vulnerability of these services to environmental change such as forest fragmentation are still poorly understood. We present spatial modeling of the generation of ecosystem services in a human-dominated landscape where forest habitat patches, protected by local taboos, are located in a matrix of cultivated land in southern Madagascar. Two ecosystem services dependent on the forest habitats were addressed: (1) crop pollination services by wild and semidomesticated bees (Apoidea), essential for local crop production of, for example, beans, and (2) seed dispersal services based on the presence of ring tailed lemurs (Lemur catta). We studied the vulnerability of these ecosystem services to a plausible scenario of successive destruction of the smallest habitat patches. Our results indicate that, in spite of the fragmented nature of the landscape, the fraction of the landscape presently covered by both crop pollination and seed dispersal services is surprisingly high. It seems that the taboo system, though indirectly and unintentionally, contributes to upholding the generation of these services by protecting the forest patches. Both services are, however, predicted to be very vulnerable to the successive removal of small patches. For crop pollination, the rate of decrease in cover was significant even when only the smallest habitat patches were removed. The capacity for seed dispersal across the landscape displayed several thresholds with habitat patch removal. Our results suggest that, in order to maintain capacity for seed dispersal across the landscape and crop pollination cover in southern Androy, the geographical location of the remaining forest patches is more crucial than their size. We argue that in heavily fragmented production landscapes, small forest patches should increasingly be viewed as essential for maintaining ecosystem services, such as agricultural production, and also should be considered in the ongoing process of tripling the area of protected habitats in Madagascar. PMID- 16711036 TI - Interactive effects of natural and human disturbances on vegetation dynamics across landscapes. AB - Accurate measures of human effects on landscape processes require consideration of both the direct impacts from human activities and the indirect consequences of the interactions between humans and the landscape. This is particularly evident in systems experiencing regular natural disturbances such as in the mountainous areas of southwestern China, where the remaining population of giant pandas (Ailuropoda melanoleuca) is supported. Here the spatiotemporal patterns of human impacts, forests, and bamboo episodic die-offs combine to determine the distribution of panda habitat. To study the complex interactions of humans and landscapes, we developed an integrated spatiotemporally explicit model of household activities, natural vegetation dynamics, and their impacts on panda habitat. Using this model we examined the direct consequences of local fuelwood collection and household creation on areas of critical giant panda habitat and the indirect impacts when coupled with vegetation dynamics. Through simulations, we found that over the next 30 years household impacts would result in the loss of up to 30% of the habitat relied on by pandas during past bamboo die-offs. The accumulation and spatial distribution of household impacts would also have a considerable indirect influence on the spatial distribution of understory bamboo. While human impacts influence both bamboo die-off and regeneration, over 19% of pre-existing low-elevation bamboo habitat may be lost following an episodic die off depending on the severity of the impacts and timing of the die-offs. Our study showed not only the importance of the spatial distribution of direct household impacts on habitat, but also the far-reaching effects of the indirect interactions between humans and the landscapes they are modifying. PMID- 16711037 TI - Tropical forest restoration: tree islands as recruitment foci in degraded lands of Honduras. AB - Tropical forest recovery in pastures is slowed by a number of biotic and abiotic factors, including a lack of adequate seed dispersal and harsh microclimatic extremes. Accordingly, methods to accelerate forest recovery must address multiple impediments. Here, we evaluated the ability of "tree islands" to serve as "recruitment foci" in a two-year study at three sites in northern Honduras. Islands of three sizes (64, 16, and 4 m2) and at two distances to secondary forest (20 and 50 m) were created by planting 2 m tall vegetative stakes of two native species: Gliricidia sepium (Fabaceae) and Bursera simaruba (Burseraceae), each in monoculture. Open-pasture "islands" of equal sizes served as controls. Tree islands reduced temperature and light (PAR) extremes as compared to open pasture, creating a microenvironment more favorable to seedling establishment. Seed-dispersing birds (quantified at one site only) showed an overwhelming preference for islands; 160 visits were recorded to islands compared with one visit to open pasture. Additionally, frugivores visited large islands more often, and for longer time periods, than small islands, thereby increasing the likelihood of a dispersal event there. In total, 144 140 seeds belonging to 186 species were collected in islands; more than 80% were grasses. Tree islands increased zoochorous tree seed rain; seed density and species richness were greater in tree islands than in open pasture, and large islands had greater seed density than smaller islands (Gliricidia only), suggesting that they are more effective for restoration. Distance to forest did not affect seed rain. A total of 543 seedlings and 41 species established in islands; > 85% were zoochorous. Seedling density did not differ among treatments (mean 0.2 seedlings/m2 for islands vs. 0.1 seedlings/m2 for pasture), although an increasing trend in tree islands over the course of two years suggests that seedling recruitment is accelerated there. Lastly, similar seedling densities were censused in the 1 m perimeter surrounding islands, suggesting that islands can expand outward into pasture. Planting vegetative stakes to create tree islands in pastures accelerates forest recovery by overcoming a number of impediments, and presents a simple, broadly applicable alternative for facilitating forest regeneration in abandoned pastures. PMID- 16711038 TI - Nitrogen spatial heterogeneity influences diversity following restoration in a ponderosa pine forest, Montana. AB - The resource heterogeneity hypothesis (RHH) is frequently cited in the ecological literature as an important mechanism for maintaining species diversity. The RHH has rarely been evaluated in the context of restoration ecology in which a commonly cited goal is to restore diversity. In this study we focused on the spatial heterogeneity of total inorganic nitrogen (TIN) following restoration treatments in a ponderosa pine (Pinus ponderosa)/Douglas-fir (Pseudotsuga menziesii) forest in western Montana, USA. Our objective was to evaluate relationships between understory species richness and TIN heterogeneity following mechanical thinning (thin-only), prescribed burning (burn-only), and mechanical thinning with prescribed burning (thin/burn) to discern the ecological and management implications of these restoration approaches. We employed a randomized block design, with three 9-ha replicates of each treatment and an untreated control. Within each treatment, we randomly established a 20 x 50 m (1000 m2) plot in which we measured species richness across the entire plot and in 12 1 m(2) quadrats randomly placed within each larger plot. Additionally, we measured TIN from a grid consisting of 112 soil samples (0-5 cm) in each plot and computed standard deviations as a measure of heterogeneity. We found a correlation between the net increase in species richness and the TIN standard deviations one and two years following restoration treatments, supporting RHH. Using nonmetric multidimensional scaling ordination and chi-squared analysis, we found that high and low TIN quadrats contained different understory communities in 2003 and 2004, further supporting RHH. A comparison of restoration treatments demonstrated that thin/burn and burn-only treatments created higher N heterogeneity relative to the control. We also found that within prescribed burn treatments, TIN heterogeneity was positively correlated with fine-fuel consumption, a variable reflecting burn severity. These findings may lead to more informed restoration decisions that consider treatment effects on understory diversity in ponderosa pine/Douglas-fir ecosystems. PMID- 16711039 TI - Species and rotation frequency influence soil nitrogen in simplified tropical plant communities. AB - Among the many factors that potentially influence the rate at which nitrogen (N) becomes available to plants in terrestrial ecosystems are the identity and diversity of species composition, frequency of disturbance or stand turnover, and time. Replicated suites of investigator-designed communities afforded an opportunity to examine the effects of those factors on net N mineralization over a 12-year period. The communities consisted of large-stature perennial plants, comprising three tree species (Hyeronima alchorneoides, Cedrela odorata, and Cordia alliodora), a palm (Euterpe oleracea), and a large, perennial herb (Heliconia imbricata). Trees were grown in monoculture and in combination with the other two life-forms; tree monocultures were subjected to rotations of one or four years, or like the three-life-form systems, left uncut. The work was conducted on fertile soil in the humid lowlands of Costa Rica, a site with few abiotic constraints to plant growth. Rates of net N mineralization and nitrification were high, typically in the range of 0.2-0.8 microg x g(1) x d(-1), with net nitrification slightly higher than net mineralization, indicating preferential uptake of ammonium (NH4+) by plants and microbes. Net rates of N mineralization were about 30% lower in stands of one of the three tree species, Hyeronima, than in stands of the other two. Contrary to expectations, short rotation management (one or four years) resulted in higher net rates of N mineralization than in uncut stands, whether the latter were composed of a single tree species or a combination of life-forms. Neither additional species richness nor replenishment of leached N augmented mineralization rates. The net rate at which N was supplied tended to be lowest in stands where demand for N was highest. Careful choice of species, coupled with low frequency of disturbance, can lead to maintenance of N within biomass and steady rates of within-system circulation, whereas pulses, whether caused by cutting and replanting or by the phenological traits of the species selected or combined, subject N supplies to leaching loss. PMID- 16711040 TI - A structural equation model analysis of postfire plant diversity in California shrublands. AB - This study investigates patterns of plant diversity following wildfires in fire prone shrublands of California, seeks to understand those patterns in terms of both local and landscape factors, and considers the implications for fire management. Ninety study sites were established following extensive wildfires in 1993, and 1000-m(2) plots were used to sample a variety of parameters. Data on community responses were collected for five years following fire. Structural equation modeling (SEM) was used to relate plant species richness to plant abundance, fire severity, abiotic conditions, within-plot heterogeneity, stand age, and position in the landscape. Temporal dynamics of average richness response was also modeled. Richness was highest in the first year following fire, indicating postfire enhancement of diversity. A general decline in richness over time was detected, with year-to-year variation attributable to annual variations in precipitation. Peak richness in the landscape was found where (1) plant abundance was moderately high, (2) within-plot heterogeneity was high, (3) soils were moderately low in nitrogen, high in sand content, and with high rock cover, (4) fire severity was low, and (5) stands were young prior to fire. Many of these characteristics were correlated with position in the landscape and associated conditions. We infer from the SEM results that postfire richness in this system is strongly influenced by local conditions and that these conditions are, in turn, predictably related to landscape-level conditions. For example, we observed that older stands of shrubs were characterized by more severe fires, which were associated with a low recovery of plant cover and low richness. These results may have implications for the use of prescribed fire in this system if these findings extrapolate to prescribed burns as we would expect. PMID- 16711041 TI - Fuel breaks affect nonnative species abundance in Californian plant communities. AB - We evaluated the abundance of nonnative plants on fuel breaks and in adjacent untreated areas to determine if fuel treatments promote the invasion of nonnative plant species. Understanding the relationship between fuel treatments and nonnative plants is becoming increasingly important as federal and state agencies are currently implementing large fuel treatment programs throughout the United States to reduce the threat of wildland fire. Our study included 24 fuel breaks located across the State of California. We found that nonnative plant abundance was over 200% higher on fuel breaks than in adjacent wildland areas. Relative nonnative cover was greater on fuel breaks constructed by bulldozers (28%) than on fuel breaks constructed by other methods (7%). Canopy cover, litter cover, and duff depth also were significantly lower on fuel breaks constructed by bulldozers, and these fuel breaks had significantly more exposed bare ground than other types of fuel breaks. There was a significant decline in relative nonnative cover with increasing distance from the fuel break, particularly in areas that had experienced more numerous fires during the past 50 years, and in areas that had been grazed. These data suggest that fuel breaks could provide establishment sites for nonnative plants, and that nonnatives may invade surrounding areas, especially after disturbances such as fire or grazing. Fuel break construction and maintenance methods that leave some overstory canopy and minimize exposure of bare ground may be less likely to promote nonnative plants. PMID- 16711042 TI - Variation in resource availability changes the impact of invasive thistles on native bunchgrasses. AB - The threat posed by invasive nonnative plants to native plant populations is one of the largest challenges facing both conservation biology and restoration ecology. California has been highly impacted by invaders, although many relict stands of native plants are found on shallow, rocky soils with limited resources. The abiotic conditions of these sites may strongly influence the performance of an invasive plant and its effect on resident native species. In addition, the maturity of native plants in these sites may modulate an invader's impact; larger, well-established plants may be better able to resist invaders. In this study we examined how the impact of an invasive thistle (Centaurea solstitialis) on a native perennial bunchgrass (Nassella pulchra) changed in response to variation in soil depth, soil water availability, and bunchgrass maturity. We measured plant performance in terms of survival, growth, reproduction, and predawn water potential. We found that soil depth, water availability, and bunchgrass maturity acted in concert to influence the impact of the invasive thistle on the native bunchgrass. Both species performed better in deep soils, especially during dry years. The combination of shallow soil and low water availability reduced C. solstitialis performance and ameliorated its negative effect on N. pulchra growth and reproduction. Higher water availability resulted in a stronger negative effect of C. solstitialis on N. pulchra in both shallow and deep soils. However, as N. pulchra matured and increased in size, we saw a steady decline in C. solstitialis growth and reproductive output. Higher water availability increased the performance of C. solstitialis in shallow soils. C. solstitialis may thus have a stronger impact on N. pulchra and be more able to invade relict stands of N. pulchra in shallow soils during high-rainfall years. However, established stands of N. pulchra appear to be more resistant to invasion by C. solstitialis as N. pulchra plants grow older and larger. PMID- 16711043 TI - Neighborhood analyses of canopy tree competition along environmental gradients in New England forests. AB - We use permanent-plot data from the USDA Forest Service's Forest Inventory and Analysis (FIA) program for an analysis of the effects of competition on tree growth along environmental gradients for the 14 most abundant tree species in forests of northern New England, USA. Our analysis estimates actual growth for each individual tree of a given species as a function of average potential diameter growth modified by three sets of scalars that quantify the effects on growth of (1) initial target tree size (dbh), (2) local environmental conditions, and (3) crowding by neighboring trees. Potential growth of seven of the 14 species varied along at least one of the two environmental axes identified by an ordination of relative abundance of species in plots. The relative abundances of a number of species were significantly displaced from sites where they showed maximum potential growth. In all of these cases, abundance was displaced to the more resource-poor end of the environmental gradient (either low fertility or low moisture). The pattern was most pronounced among early successional species, whereas late-successional species reached their greatest abundance on sites where they also showed the highest growth in the absence of competition. The analysis also provides empirical estimates of the strength of intraspecific and interspecific competitive effects of neighbors. For all but one of the species, our results led us to reject the hypothesis that all species of competitors have equivalent effects on a target species. Most of the individual pairwise interactions were strongly asymmetric. There was a clear competitive hierarchy among the four most shade-tolerant species, and a separate competitive hierarchy among the shade-intolerant species. Our results suggest that timber yield following selective logging will vary dramatically depending on the configuration of the residual canopy, because of interspecific variation in the magnitude of both the competitive effects of different species of neighbors and the competitive responses of different species of target trees to neighbors. The matrix of competition coefficients suggests that there may be clear benefits in managing for specific mixtures of species within local neighborhoods within stands. PMID- 16711044 TI - Urbanization increases grassland carbon pools: effects of landscaping in Colorado's front range. AB - During the past few decades, urban and suburban developments have grown at unprecedented rates and extents with unknown consequences for ecosystem function. Carbon pools of soil and vegetation on landscaped properties were examined in the Front Range of Colorado, USA, in order to characterize vegetation and soils found in urban green spaces; analyze their aboveground biomass, vegetative C storage, and soil C storage; and compare these suburban ecosystem properties to their counterparts in native grassland and cultivated fields. Anthropogenic activities leave clear signatures on all three C compartments measured. Management level dominates the response of grass production, biomass, and N tissue concentration. This, in turn, influences the amount of C and N both stored in and harvested from sites. The site age dominates the amount of woody biomass as well as soil C and N. Soil texture only secondarily affects total soil carbon and total bulk density. Established urban green spaces harbor larger C pools, more than double in some cases, than native grasslands or agricultural fields on a per-area basis. Lawn grass produces more biomass and stores more C than local prairie or agricultural fields. Introduced woody vegetation comprises a substantial C pool in urban green spaces and represents a new ecosystem feature. After an initial decrease with site development, soil organic carbon (SOC) pools surpass those in grasslands within two decades. In addition to the marked increase of C pools through time, a shift in storage from belowground to aboveground occurs. Whereas grasslands store approximately 90% of C belowground, urban green spaces store a decreasing proportion of the total C belowground in soils through time, reaching approximately 70% 30-40 years after construction. Despite the substantial increase in C pools in this urban area, it is important to recognize that this shift is distinct from C sequestration since it does not account for a total C budget, including increased anthropogenic C emissions from these sites. PMID- 16711045 TI - Pastoralists' responses to variation of rangeland resources in time and space. AB - We explore the response of pastoralists to rangeland resource variation in time and space, focusing on regions where high variation makes it unlikely that an economically viable herd can be maintained on a single management unit. In such regions, the need to move stock to find forage in at least some years has led to the evolution of nomadism and transhumance, and reciprocal grazing agreements among the holders of common-property rangeland. The role of such informal institutions in buffering resource variation is well documented in some Asian and African rangelands, but in societies with formally established private-property regimes, where we focus, such institutions have received little attention. We examine agistment networks, which play an important role in buffering resource variation in modern-day Australia. Agistment is a commercial arrangement between pastoralists who have less forage than they believe they require and pastoralists who believe they have more. Agistment facilitates the movement of livestock via a network based largely on trust. We are concerned exclusively with the link between the characteristics of biophysical variation and human aspects of agistment networks, and we developed a model to test the hypothesis that such a link could exist. Our model builds on game theory literature, which explains cooperation between strangers based on the ability of players to learn whom they can trust. Our game is played on a highly stylized landscape that allows us to control and isolate the degree of spatial variation and spatial covariation. We found that agistment networks are more effective where spatial variation in resource availability is high, and generally more effective when spatial covariation is low. Policy design that seeks to work with existing social networks in rangelands has potential, but this potential varies depending on localized characteristics of the biophysical variability. PMID- 16711046 TI - Demography of northern flying squirrels informs ecosystem management of western interior forests. AB - We studied northern flying squirrel (Glaucomys sabrinus) demography in the eastern Washington Cascade Range to test hypotheses about regional and local abundance patterns and to inform managers of the possible effects of fire and fuels management on flying squirrels. We quantified habitat characteristics and squirrel density, population trends, and demography in three typical forest cover types over a four-year period. We had 2034 captures of flying squirrels over 41 000 trap nights from 1997 through 2000 and marked 879 squirrels for mark recapture population analysis. Ponderosa pine (Pinus ponderosa) forest appeared to be poorer habitat for flying squirrels than young or mature mixed-conifer forest. About 35% fewer individuals were captured in open pine forest than in dry mixed-conifer Douglas-fir (Pseudotsuga menziesii) and grand fir (Abies grandis) forests. Home ranges were 85% larger in pine forest (4.6 ha) than in mixed conifer forests (2.5 ha). Similarly, population density (Huggins estimator) in ponderosa pine forest was half (1.1 squirrels/ha) that of mixed-conifer forest (2.2 squirrels/ha). Tree canopy cover was the single best correlate of squirrel density (r = 0.77), with an apparent threshold of 55% canopy cover separating stands with low- from high-density populations. Pradel estimates of annual recruitment were lower in open pine (0.28) than in young (0.35) and mature (0.37) forest. High recruitment was most strongly associated with high understory plant species richness and truffle biomass. Annual survival rates ranged from 45% to 59% and did not vary among cover types. Survival was most strongly associated with understory species richness and forage lichen biomass. Maximum snow depth had a strong negative effect on survival. Rate of per capita increase showed a density-dependent response. Thinning and prescribed burning in ponderosa pine and dry mixed conifer forests to restore stable fire regimes and forest structure might reduce flying squirrel densities at stand levels by reducing forest canopy, woody debris, and the diversity or biomass of understory plants, truffles, and lichens. Those impacts might be ameliorated by patchy harvesting and the retention of large trees, woody debris, and mistletoe brooms. Negative stand level impacts would be traded for increased resistance and resilience of dry forest landscapes to now-common, large-scale stand replacement fires. PMID- 16711047 TI - Recruitment of Hexagenia mayfly nymphs in western Lake Erie linked to environmental variability. AB - After a 40-year absence caused by pollution and eutrophication, burrowing mayflies (Hexagenia spp.) recolonized western Lake Erie in the mid 1990s as water quality improved. Mayflies are an important food resource for the economically valuable yellow perch fishery and are considered to be major indicator species of the ecological condition of the lake. Since their reappearance, however, mayfly populations have suffered occasional unexplained recruitment failures. In 2002, a failure of fall recruitment followed an unusually warm summer in which western Lake Erie became temporarily stratified, resulting in low dissolved oxygen levels near the lake floor. In the present study, we examined a possible link between Hexagenia recruitment and periods of intermittent stratification for the years 1997 2002. A simple model was developed using surface temperature, wind speed, and water column data from 2003 to predict stratification. The model was then used to detect episodes of stratification in past years for which water column data are unavailable. Low or undetectable mayfly recruitment occurred in 1997 and 2002, years in which there was frequent or extended stratification between June and September. Highest mayfly reproduction in 2000 corresponded to the fewest stratified periods. These results suggest that even relatively brief periods of stratification can result in loss of larval mayfly recruitment, probably through the effects of hypoxia. A trend toward increasing frequency of hot summers in the Great Lakes region could result in recurrent loss of mayfly larvae in western Lake Erie and other shallow areas in the Great Lakes. PMID- 16711048 TI - Emergence cues of a mayfly in a high-altitude stream ecosystem: potential response to climate change. AB - To understand the consequences of human accelerated environmental change, it is important to document the effects on natural populations of an increasing frequency of extreme climatic events. In stream ecosystems, recent climate change has resulted in extreme variation in both thermal and hydrological regimes. From 2001 to 2004, a severe drought in western United States corresponded with earlier emergence of the adult stage of the high-altitude stream mayfly, Baetis bicaudatus. Using a long-term database from a western Colorado stream, the peak emergence date of this mayfly population was predicted by both the magnitude and date of peak stream flow, and by the mean daily water temperature, suggesting that Baetis may respond to declining stream flow or increasing water temperature as proximate cues for early metamorphosis. However, in a one-year survey of multiple streams from the same drainage basin, only water temperature predicted spatial variation in the onset of emergence of this mayfly. To decouple the effects of temperature and flow, we separately manipulated these factors in flow through microcosms and measured the timing of B. bicaudatus metamorphosis to the adult stage. Mayflies emerged sooner in a warmed-water treatment than an ambient water treatment; but reducing flow did not accelerate the onset of mayfly emergence. Nonetheless, using warming temperatures to cue metamorphosis enables mayflies to time their emergence during the descending limb of the hydrograph when oviposition sites (protruding rocks) are becoming available. We speculate that large-scale climate changes involving warming and stream drying could cause significant shifts in the timing of mayfly metamorphosis, thereby having negative effects on populations that play an important role in stream ecosystems. PMID- 16711049 TI - Arthropod food web restoration following removal of an invasive wetland plant. AB - Restoration of habitats impacted by invasive plants is becoming an increasingly important tool in the management of native biodiversity, though most studies do not go beyond monitoring the abundance of particular taxonomic groups, such as the return of native vegetation. Yet, the reestablishment of trophic interactions among organisms in restored habitats is equally important if we are to monitor and understand how ecosystems recover. This study examined whether food web interactions among arthropods (as inferred by abundance of naturally occurring stable isotopes of C [delta13C] and N [delta15N]) were reestablished in the restoration of a coastal Spartina alterniflora salt marsh that had been invaded by Phragmites australis. From patterns of C and N stable isotopes we infer that trophic interactions among arthropods in the native salt marsh habitats are characterized by reliance on the dominant marsh plant Spartina as a basal resource. Herbivores such as delphacid planthoppers and mirid bugs have isotope signatures characteristic of Spartina, and predatory arthropods such as dolicopodid flies and spiders likewise have delta13C and delta15N signatures typical of Spartina-derived resources (approximately -13 per thousand and 10 per thousand, respectively). Stable isotope patterns also suggest that the invasion of Phragmites into salt marshes and displacement of Spartina significantly alter arthropod food web interactions. Arthropods in Phragmites-dominated sites have delta13C isotope values between -18 per thousand and -20 per thousand, suggesting reliance on detritus and/or benthic microalgae as basal resources and not on Phragmites, which has a delta13C approximately -26 per thousand. Since most Phragmites herbivores are either feeding internally or are rare transients from nearby Spartina, these resources do not provide significant prey resources for other arthropod consumers. Rather, predator isotope signatures in the invaded habitats indicate dependence on detritus/algae as basal resources instead of the dominant vegetation. The reestablishment of Spartina after removal of Phragmites, however, not only returned species assemblages typical of reference (uninvaded) Spartina, but stable isotope signatures suggest that the trophic interactions among the arthropods were also similar in reestablished habitats. Specifically, both herbivores and predators showed characteristic Spartina signatures, suggesting the return of the original grazer-based food web structure in the restored habitats. PMID- 16711050 TI - Complex responses within a desert bee guild (Hymenoptera: Apiformes) to urban habitat fragmentation. AB - Urbanization within the Tucson Basin of Arizona during the past 50+ years has fragmented the original desert scrub into patches of different sizes and ages. These remnant patches and the surrounding desert are dominated by Larrea tridentata (creosote bush), a long-lived shrub whose flowers are visited by > 120 native bee species across its range. Twenty-one of these bee species restrict their pollen foraging to L. tridentata. To evaluate the response of this bee fauna to fragmentation, we compared species incidence and abundance patterns for the bee guild visiting L. tridentata at 59 habitat fragments of known size (0.002 5 ha) and age (up to 70 years), and in adjacent desert. The 62 bee species caught during this study responded to fragmentation heterogeneously and not in direct relation to their abundance or incidence in undisturbed desert. Few species found outside the city were entirely absent from urban fragments. Species of ground nesting L. tridentata specialists were underrepresented in smaller fragments and less abundant in the smaller and older fragments. In contrast, cavity-nesting bees (including one L. tridentata specialist) were overrepresented in the habitat fragments, probably due to enhanced nesting opportunities available in the urban matrix. Small-bodied bee species were no more likely than larger bodied species to be absent from the smaller fragments. The introduced European honey bee, Apis mellifera, was a minor faunal element at > 90% of the fragments and exerted little if any influence on the response of native bee species to fragmentation. Overall, bee response to urban habitat fragmentation was best predicted by ecological traits associated with nesting and dietary breadth. Had species been treated as individual units in the analyses, or pooled together into one analysis, these response patterns may not have been apparent. Pollination interactions with this floral host are probably not adversely affected in this system because of its longevity and ability to attract diverse pollinators but will demand careful further study to understand. PMID- 16711051 TI - Community response patterns: evaluating benthic invertebrate composition in metal polluted streams. AB - Human activities are modifying the condition and character of ecosystems at a rapid rate. Because of these rapid changes, questions concerning how ecosystems and their assemblages respond to anthropogenic stressors have been of general interest. Accurate prediction of assemblage composition in ecosystems with anthropogenic degradation requires that we assess both how assemblages respond to stressors and the generality of the responses. We ask whether assemblage composition among stream sites becomes more similar after exposure to a common stressor. Using data from biological monitoring programs in the southern Rocky Mountain ecoregion of Colorado and in West Virginia, we compare benthic invertebrate similarity and assemblage composition among sites having different levels (background, low, medium, and high) of heavy-metal pollution. Invertebrate assemblages were most similar within the background metal category, and similarity was progressively lower in low, medium, and high metal categories. An analysis of the frequency of occurrence of genera within metal categories reveals taxonomic shifts that conform to expectations based on metal tolerance of benthic invertebrates. However, different metal-tolerant genera were found at different metal-impacted sites, suggesting that local abiotic and biotic processes may influence the identity of the metal-tolerant genera that become established in polluted sites. Low community similarity in the medium and high-metal categories suggests that accurate prediction of assemblage composition at impacted sites may be challenging. PMID- 16711052 TI - Population growth rate and carrying capacity for springtails Folsomia candida exposed to ivermectin. AB - Forecasting the effects of stressors on the dynamics of natural populations requires assessment of the joint effects of a stressor and population density on the population response. The effects can be depicted as a contour map in which the population response, here assessed by population growth rate, varies with stress and density in the same way that the height of land above sea level varies with latitude and longitude. We present the first complete map of this type using as our model Folsomia candida exposed to five different concentrations of the widespread anthelmintic veterinary medicine ivermectin in replicated microcosm experiments lasting 49 days. The concentrations of ivermectin in yeast were 0.0, 6.8, 28.8, 66.4, and 210.0 mg/L wet weight. Increasing density and chemical concentration both significantly reduced the population growth rate of Folsomia candida, in part through effects on food consumption and fecundity. The interaction between density and ivermectin concentration was "less-than additive," implying that at high density populations were able to compensate for the effects of the chemical. This result demonstrates that regulatory protocols carried out at low density (as in most past experiments) may seriously overestimate effects in the field, where densities are locally high and populations are resource limited (e.g., in feces of livestock treated with ivermectin). PMID- 16711053 TI - Interactive effects of prey and p,p'-DDE on burrowing owl population dynamics. AB - We used population models to explore the effects of the organochlorine contaminant p,p'-DDE and fluctuations in vole availability on the population dynamics of Burrowing Owls (Athene cunicularia). Previous work indicated an interaction between low biomass of voles in the diet and moderate levels of p,p' DDE in Burrowing Owl eggs that led to reproductive impairment. We constructed periodic and stochastic matrix models that incorporated three vole population states observed in the field: average, peak, and crash years. We modeled varying frequencies of vole crash years and a range of impairment of owl demographic rates in vole crash years. Vole availability had a greater impact on owl population growth rate than did reproductive impairment if vole populations peaked and crashed frequently. However, this difference disappeared as the frequency of vole crash years declined to once per decade. Fecundity, the demographic rate most affected by p,p'-DDE, had less impact on population growth rate than adult or juvenile survival. A life table response experiment of time invariant matrices for average, peak, and crash vole conditions showed that low population growth under vole crash conditions was due to low adult and juvenile survival rates, whereas the extremely high population growth under vole peak conditions was due to increased fecundity. Our results suggest that even simple models can provide useful insights into complex ecological interactions. This is particularly valuable when temporal or spatial scales preclude manipulative experimental work in the field or laboratory. PMID- 16711054 TI - Albatross species demonstrate regional differences in North Pacific marine contamination. AB - Recent concern about negative effects on human health from elevated organochlorine and mercury concentrations in marine foods has highlighted the need to understand temporal and spatial patterns of marine pollution. Seabirds, long-lived pelagic predators with wide foraging ranges, can be used as indicators of regional contaminant patterns across large temporal and spatial scales. Here we evaluate contaminant levels, carbon and nitrogen stable isotope ratios, and satellite telemetry data from two sympatrically breeding North Pacific albatross species to demonstrate that (1) organochlorine and mercury contaminant levels are significantly higher in the California Current compared to levels in the high latitude North Pacific and (2) levels of organochlorine contaminants in the North Pacific are increasing over time. Black-footed Albatrosses (Phoebastria nigripes) had 370-460% higher organochlorine (polychlorinated biphenyls [PCBs], dichlorodiphenyltrichloroethanes [DDTs]) and mercury body burdens than a closely related species, the Laysan Albatross (P. immutabilis), primarily due to regional segregation of their North Pacific foraging areas. PCBs (the sum of the individual PCB congeners analyzed) and DDE concentrations in both albatross species were 130-360% higher than concentrations measured a decade ago. Our results demonstrate dramatically high and increasing contaminant concentrations in the eastern North Pacific Ocean, a finding relevant to other marine predators, including humans. PMID- 16711055 TI - Habitat edge, land management, and rates of brood parasitism in tallgrass prairie. AB - Bird populations in North America's grasslands have declined sharply in recent decades. These declines are traceable, in large part, to habitat loss, but management of tallgrass prairie also has an impact. An indirect source of decline potentially associated with management is brood parasitism by the Brown-headed Cowbird (Molothrus ater), which has had substantial negative impacts on many passerine hosts. Using a novel application of regression trees, we analyzed an extensive five-year set of nest data to test how management of tallgrass prairie affected rates of brood parasitism. We examined seven landscape features that may have been associated with parasitism: presence of edge, burning, or grazing, and distance of the nest from woody vegetation, water, roads, or fences. All five grassland passerines that we included in the analyses exhibited evidence of an edge effect: the Grasshopper Sparrow (Ammodramus savannarum), Henslow's Sparrow (A. henslowii), Dickcissel (Spiza americana), Red-winged Blackbird (Agelaius phoeniceus), and Eastern Meadowlark (Sturnella magna). The edge was represented by narrow strips of woody vegetation occurring along roadsides cut through tallgrass prairie. The sparrows avoided nesting along these woody edges, whereas the other three species experienced significantly higher (1.9-5.3x) rates of parasitism along edges than in prairie. The edge effect could be related directly to increase in parasitism rate with decreased distance from woody vegetation. After accounting for edge effect in these three species, we found evidence for significantly higher (2.5-10.5x) rates of parasitism in grazed plots, particularly those burned in spring to increase forage, than in undisturbed prairie. Regression tree analysis proved to be an important tool for hierarchically parsing various landscape features that affect parasitism rates. We conclude that, on the Great Plains, rates of brood parasitism are strongly associated with relatively recent road cuts, in that edge effects manifest themselves through the presence of trees, a novel habitat component in much of the tallgrass prairie. Grazing is also a key associate of increased parasitism. Areas managed with prescribed fire, used frequently to increase forage for grazing cattle, may experience higher rates of brood parasitism. Regardless, removing trees and shrubs along roadsides and refraining from planting them along new roads may benefit grassland birds. PMID- 16711056 TI - Additive effects of vertebrate predators on insects in a Puerto Rican coffee plantation. AB - A variety of studies have established the value of shaded coffee plantations as habitat for birds. While the value of birds as biological controls in coffee has received some attention, the interactions between birds and other predators of insects have not been tested. We used exclosures to examine the effects of vertebrate predators on the arthropods associated with coffee, in particular the coffee leafminer (Leucoptera coffeella) and the flatid planthopper Petrusa epilepsis, in a shaded coffee plantation in Puerto Rico. We used a 2 x 2 factorial design with four treatments: exclusion of birds, lizards, birds and lizards, and control (no exclusion). Abundance of insects > 5 mm increased when birds or both birds and lizards were removed. Birds and lizards had an additive effect for insects < 5 mm and for all insects combined. Coffee leafminers showed a weak response to removal of predators while planthopper abundance increased significantly in the absence of avian predators. Arthropod predators and parasitoids did not differ significantly between treatments. Our findings suggest that vertebrate insectivores have an additive effect on insects in coffee and may help control abundances of some coffee pests. Equally important, we present evidence suggesting that they do not interfere with other known natural enemies of coffee pests. PMID- 16711057 TI - Associations between the Alaska Steller sea lion decline and commercial fisheries. AB - The Steller sea lion (SSL) population in Alaska was listed as threatened under the Endangered Species Act in 1990. At that time, several procedural restrictions were placed on the commercial fisheries of the region in an effort to reduce the potential for human-induced mortality on sea lions. Several years have elapsed since these restrictions were put into place, and questions about their efficacy remain. In an effort to determine whether or not fisheries management measures have helped the SSL population to recover, estimates of the fishing activity of the Bering Sea/Gulf of Alaska commercial fisheries in the vicinity of individual SSL rookeries and SSL population trends at those rookeries were made using data from the National Marine Fisheries Service (NMFS) Fisheries Observer Program and Steller Sea Lion Adult Count Database. Fisheries data from 1976-2000 were analyzed in relation to SSL population counts from 1956-2001 at 32 rookeries from the endangered western stock. Linear regression on the principal components of the fisheries data show that a positive correlation exists between several metrics of historical fishing activity and the SSL population decline. The relationship is less consistent after 1991, supporting a hypothesis that management measures around some of the rookeries have been effective in moderating the localized effects of fishing activity on SSL. PMID- 16711058 TI - When r-selection may not predict introduced-species proliferation: predation of a nonnative oyster. AB - Predicting outcomes of species introductions may be enhanced by integrating life history theory with results of contained experiments that compare ecological responses of exotic and analogue native species to dominant features of the recipient environment. An Asian oyster under consideration for introduction to the Chesapeake Bay, USA, the rapidly growing Suminoe oyster (Crassostrea ariakensis), may not be as successful an invader as its r-selected life history suggests if the trade-off for rapid growth and maturation is lower investment in defenses against blue crab (Callinectes sapidus) predation than the native Eastern oyster (Crassostrea virginica). In laboratory trials, blue crabs simultaneously offered equal numbers of Suminoe and Eastern oysters consumed more nonnatives, irrespective of whether the crabs had previous experience with Suminoe oysters as prey. Satiated blue crabs consumed nearly three times as many Suminoe oysters as Eastern oysters of 25-mm shell height, and eight times as many of 35-mm shell height. Despite blue crabs consuming small (30 mm) Suminoe oysters at twice the rate of large (40 mm) Suminoe oysters, when 40-mm Suminoe were paired with 30-mm Eastern oysters, seven times as many of the larger (Suminoe) oysters were consumed. The greater susceptibility of C. ariakensis than C. virginica to blue crab predation appears to be based upon the biomechanics of shell strength rather than active selection of a more attractive food. Much less force was required to crush shells of Suminoe than Eastern oysters of similar shell height. Tissue transplant experiments demonstrated greater predation on oyster tissues in weaker C. ariakensis shells independent of tissue identity, and duration of handling time before rejection of C. virginica exceeded the time to crush C. ariakensis. These results, coupled with the present importance of blue crab predation in limiting recovery of native Eastern oysters, imply a role for blue crabs in inhibiting Suminoe oysters, if introduced, from attaining high adult densities required to restore a fishery, provide appreciable reef habitat, and reduce turbidity through filtration. Thus, in high-predation environments, allocation of resources to rapid growth and development rather than to predation defenses reflects a life-history trade-off that may promote early stages of invasion, yet prevent attainment of dense adult populations. PMID- 16711059 TI - Density-dependent habitat selection and performance by a large mobile reef fish. AB - Many exploited reef fish are vulnerable to overfishing because they concentrate over hard-bottom patchy habitats. How mobile reef fish use patchy habitat, and the potential consequences on demographic parameters, must be known for spatially explicit population dynamics modeling, for discriminating essential fish habitat (EFH), and for effectively planning conservation measures (e.g., marine protected areas, stock enhancement, and artificial reefs). Gag, Mycteroperca microlepis, is an ecologically and economically important warm-temperate grouper in the southeastern United States, with behavioral and life history traits conducive to large-scale field experiments. The Suwannee Regional Reef System (SRRS) was built of standard habitat units (SHUs) in 1991-1993 to manipulate and control habitat patchiness and intrinsic habitat quality, and thereby test predictions from habitat selection theory. Colonization of the SRRS by gag over the first six years showed significant interactions of SHU size, spacing, and reef age; with trajectories modeled using a quadratic function for closely spaced SHUs (25 m) and a linear model for widely spaced SHUs (225 m), with larger SHUs (16 standardized cubes) accumulating significantly more gag faster than smaller 4 cube SHUs (mean = 72.5 gag/16-cube SHU at 225-m spacing by year 6, compared to 24.2 gag/4-cube SHU for same spacing and reef age). Residency times (mean = 9.8 mo), indicative of choice and measured by ultrasonic telemetry (1995-1998), showed significant interaction of SHU size and spacing consistent with colonization trajectories. Average relative weight (W(r)) and incremental growth were greater on smaller than larger SHUs (mean W(r) = 104.2 vs. 97.7; incremental growth differed by 15%), contrary to patterns of abundance and residency. Experimental manipulation of shelter on a subset of SRRS sites (2000-2001) confirmed our hypothesis that shelter limits local densities of gag, which, in turn, regulates their growth and condition. Density-dependent habitat selection for shelter and individual growth dynamics were therefore interdependent ecological processes that help to explain how patchy reef habitat sustains gag production. Moreover, gag selected shelter at the expense of maximizing their growth. Thus, mobile reef fishes could experience density-dependent effects on growth, survival, and/or reproduction (i.e., demographic parameters) despite reduced stock sizes as a consequence of fishing. PMID- 16711060 TI - The impact of exploiting grazers (Scaridae) on the dynamics of Caribbean coral reefs. AB - Coral reefs provide a number of ecosystem services including coastal defense from storms, the generation of building materials, and fisheries. It is increasingly clear that the management of reef resources requires an ecosystem approach in which extractive activities are weighed against the needs of the ecosystem and its functions rather than solely those of the fishery. Here, I use a spatially explicit simulation model of a Caribbean coral reef to examine the ecosystem requirements for grazing which is primarily conducted by parrotfishes (Scaridae). The model allows the impact of fishing grazers to be assessed in the wider context of other ecosystem processes including coral-algal competition, hurricanes, and mass extinction of the herbivorous urchin Diadema antillarum. Using a new analytical model of scarid grazing, it is estimated that parrotfishes can only maintain between 10% and 30% of a structurally complex forereef in a grazed state. Predictions from this grazing model were then incorporated into a broader simulation model of the ecosystem. Simulations predict that scarid grazing is unable to maintain high levels of coral cover (> or = 30%) when severe hurricanes occur on a decadal basis, such as occurs in parts of the northern Caribbean. However, reefs can withstand such intense disturbance when grazing is undertaken by both scarids and the urchin Diadema. Scarid grazing is predicted to allow recovery from hurricanes when their incidence falls to 20 years or less (e.g., most of Central and South America). Sensitivity analyses revealed that scarid grazing had the most acute impact on model behavior, and depletion led to the emergence of a stable, algal-dominated community state. Under conditions of heavy grazer depletion, coral cover was predicted to decline rapidly from an initial level of 30% to less than 1% within 40 years, even when hurricane frequency was low at 60 years. Depleted grazers caused a population bottleneck in juvenile corals in which algal overgrowth caused elevated levels of postsettlement mortality and resulted in a bimodal distribution of coral sizes. Several new hypotheses were generated including a region-wide change in the spatial heterogeneity of coral reefs following extinction of Diadema. The management of parrotfishes on Caribbean reefs is usually approached implicitly through no-take marine reserves. The model predicts that depletion of grazers in nonreserve areas can severely limit coral accretion. Other studies have shown that low coral accretion can reduce the structural complexity and therefore quality of the reef habitat for many organisms. A speculative yet rational inference from the model is that failure to manage scarid populations outside reserves will have a profoundly negative impact on the functioning of the reserve system and status of non-reserve reefs. PMID- 16711061 TI - Risk of Myxobolus cerebralis infection to rainbow trout in the Madison River, Montana, USA. AB - Myxobolus cerebralis, the parasite that causes salmonid whirling disease, has had detrimental effects on several salmonid populations in the Intermountain West, including the rainbow trout in the Madison River, Montana, USA. The goal of this study was to examine relationships among characteristics of the environment, Tubifex tubifex (the alternate host) populations, and rainbow trout whirling disease risk in the Madison River. Environmental characteristics were measured in side channels of the Madison River, and differences were described with a principal components analysis. The density of T. tubifex, the prevalence of infection in T. tubifex, and the density of infected T. tubifex were determined for the side channels using benthic core samples and examination of live tubificids for infection. The site-specific contribution to whirling disease risk in the side channels was determined using in situ exposures of sentinel rainbow trout. Regression analyses were used to determine correlations among these characteristics. Side channels differed in site-specific contribution to rainbow trout whirling disease risk, which was positively correlated to the density of infected T. tubifex. Side channels with fine sediments and lower water temperatures made greater site-specific contribution to whirling disease risk and had higher densities of infected T. tubifex than side channels with coarser sediments and higher temperatures. The ability to characterize areas of high whirling disease risk is essential for improving our understanding of the dynamics of M. cerebralis such that appropriate management strategies can be implemented. In addition, this study provides a model of how the disease ecology of complex aquatic parasites can be examined when the influential processes operate on different spatial scales. PMID- 16711062 TI - Did the pre-1980 use of in-stream structures improve streams? A reanalysis of historical data. AB - In the 1930s, after only three years of scientific investigation at the University of Michigan Institute for Fisheries Research, cheap labor and government-sponsored conservation projects spearheaded by the Civilian Conservation Corps allowed the widespread adoption of in-stream structures throughout the United States. From the 1940s through the 1970s, designs of in stream structures remained essentially unchanged, and their use continued. Despite a large investment in the construction of in-stream structures over these four decades, very few studies were undertaken to evaluate the impacts of the structures on the channel and its aquatic populations. The studies that were undertaken to evaluate the impact of the structures were often flawed. The use of habitat structures became an "accepted practice," however, and early evaluation studies were used as proof that the structures were beneficial to aquatic organisms. A review of the literature reveals that, despite published claims to the contrary, little evidence of the successful use of in-stream structures to improve fish populations exists prior to 1980. A total of 79 publications were checked, and 215 statistical analyses were performed. Only seven analyses provide evidence for a benefit of structures on fish populations, and five of these analyses are suspect because data were misclassified by the original authors. Many of the changes in population measures reported in early publications appear to result from changes in fishing pressure that often accompanied channel modifications. Modern evaluations of channel-restoration projects must consider the influence of fishing pressure to ensure that efforts to improve fish habitat achieve the benefits intended. My statistical results show that the traditional use of in-stream structures for channel restoration design does not ensure demonstrable benefits for fish communities, and their ability to increase fish populations should not be presumed. PMID- 16711063 TI - Using life history and persistence criteria to prioritize habitats for management and conservation. AB - In many marine and terrestrial systems, individuals of a focal species may be found in habitats that are neither essential nor of particular concern for conservation. For example, finding fish in a location does not make that location essential. This recognition begs the question of how one identifies the most important habitats for a particular species. We introduce new tools for use when prioritizing habitats for conservation and management, with application to Essential Fish Habitat (EFH). If density-dependent effects can be ignored, then elasticities of population growth rate with respect to adult survival, fecundity, and juvenile survival provide a means of identifying how susceptible the growth rate of the population is to perturbations in vital rates of particular life stages. We develop such a theory and apply it to 16 species of commercially harvested Sebastes rockfishes. We also show that the conclusions may differ significantly depending upon the estimate of mortality rate that is used. This suggests that although estimating mortality rates of fish in the field is difficult, it is crucial to do so. When density-dependent effects are important, we use a stochastic life history model to compute the moderate-term persistence of the stock, an important metric of population biology, as a function of the demographic parameters determined by the habitat. Although developed with fish in mind, the ideas here also apply to other taxa and systems. PMID- 16711064 TI - Should managed populations be monitored every year? AB - We often need to estimate the size of wild populations to determine the appropriate management action, for example, to set a harvest quota. Monitoring is usually planned under the assumption that it must be carried out at fixed intervals in time, typically annually, before the harvest quota is set. However, monitoring can be very expensive, and we should weigh the cost of monitoring against the improvement that it makes in decision making. A less costly alternative to monitoring annually is to predict the population size using a population model and information from previous surveys. In this paper, the problem of monitoring frequency is posed within a decision-theory framework. We discover that a monitoring regime that varies according to the state of the system can outperform fixed-interval monitoring. This idea is illustrated using data for a red kangaroo (Macropus rufus) population in South Australia. Whether or not one should monitor in a given year is dependent on the estimated population density in the previous year, the uncertainty in that population estimate, and past rainfall. We discover that monitoring is important when a model-based prediction of population density is very uncertain. This may occur if monitoring has not taken place for several years, or if rainfall has been above average. Monitoring is also important when prior information suggests that the population is near a critical threshold in population abundance. However, monitoring is less important when the optimal management action would not be altered by new information. PMID- 16711065 TI - Average dispersal success: linking home range, dispersal, and metapopulation dynamics to reserve design. AB - Spatially explicit models for populations are often difficult to tackle mathematically and, in addition, require detailed data on individual movement behavior that are not easily obtained. An approximation known as the "average dispersal success" provides a tool for converting complex models, which may include stage structure and a mechanistic description of dispersal, into a simple matrix model. This simpler matrix model has two key advantages. First, it is easier to parameterize from the types of empirical data typically available to conservation biologists, such as survivorship, fecundity, and the fraction of juveniles produced in a study area that also recruit within the study area. Second, it is more amenable to theoretical investigation. Here, we use the average dispersal success approximation to develop estimates of the critical reserve size for systems comprising single patches or simple metapopulations. The quantitative approach can be used for both plants and animals; however, to provide a concrete example of the technique's utility, we focus on a special case pertinent to animals. Specifically, for territorial animals, we can characterize such an estimate of minimum viable habitat area in terms of the number of home ranges that the reserve contains. Consequently, the average dispersal success framework provides a framework through which home range size, natal dispersal distances, and metapopulation dynamics can be linked to reserve design. We briefly illustrate the approach using empirical data for the swift fox (Vulpes velox). PMID- 16711066 TI - Estimating the abundance of mouse populations of known size: promises and pitfalls of new methods. AB - Knowledge of animal abundance is fundamental to many ecological studies. Frequently, researchers cannot determine true abundance, and so must estimate it using a method such as mark-recapture or distance sampling. Recent advances in abundance estimation allow one to model heterogeneity with individual covariates or mixture distributions and to derive multimodel abundance estimators that explicitly address uncertainty about which model parameterization best represents truth. Further, it is possible to borrow information on detection probability across several populations when data are sparse. While promising, these methods have not been evaluated using mark-recapture data from populations of known abundance, and thus far have largely been overlooked by ecologists. In this paper, we explored the utility of newly developed mark-recapture methods for estimating the abundance of 12 captive populations of wild house mice (Mus musculus). We found that mark-recapture methods employing individual covariates yielded satisfactory abundance estimates for most populations. In contrast, model sets with heterogeneity formulations consisting solely of mixture distributions did not perform well for several of the populations. We show through simulation that a higher number of trapping occasions would have been necessary to achieve good estimator performance in this case. Finally, we show that simultaneous analysis of data from low abundance populations can yield viable abundance estimates. PMID- 16711067 TI - Substrate modulation of osteoblast adhesion strength, focal adhesion kinase activation, and responsiveness to mechanical stimuli. AB - Osteoblast interactions with extracellular matrix (ECM) proteins are known to influence many cell functions, which may ultimately affect osseointegration of implants with the host bone tissue. Some adhesion-mediated events include activation of focal adhesion kinase, and subsequent changes in the cytoskeleton and cell morphology, which may lead to changes in adhesion strength and cell responsiveness to mechanical stimuli. In this study we examined focal adhesion kinase activation (FAK), F-actin cytoskeleton reorganization, adhesion strength, and osteoblast responsiveness to fluid shear when adhered to type I collagen (ColI), glass, poly-L-lysine (PLL), fibronectin (FN), vitronectin (VN), and serum (FBS). In general, surfaces that bind cells through integrins (FN, VN, FBS) elicited the highest adhesion strength, FAK activation, and F-actin stress fiber formation after both 15 and 60 minutes of adhesion. In contrast, cells attached through non-integrin mediated means (PLL, glass) showed the lowest FAK activation, adhesion strength, and little F-actin stress fiber formation. When subjected to steady fluid shear using a parallel plate flow chamber, osteoblasts plated on FN released significantly more PGE2 compared to those on glass. In contrast, PGE2 release of osteoblasts attached to FN or glass was not different in the absence of fluid shear, suggesting that differences in binding alone are insufficient to alter PGE2 secretion. The increased adhesion strength as well as PGE2 secretion of osteoblasts adhered via integrins may be due to increased F actin fiber formation, which leads to increased cell stiffness. PMID- 16711068 TI - Stretching short DNAs in electrolytes. AB - This paper is aimed at a combined theoretical and numerical study of the force extension relation of a short DNA molecule stretched in an electrolyte. A theoretical formula based on a recent discrete wormlike chain (WLC) model of Kierfeld et al. (Eur Phys. J. E, Vol. 14, pp.17-34, 2004) and the classical OSF mean-field theory on electrostatic stiffening of a charged polymer is numerically verified by a set of Brownian dynamics simulations based on a generalized bead rod (GBR) model incorporating long-ranged electrostatic interactions via the Debye-Hueckel potential (DH). The analysis indicates that the stretching of a short DNA can be well described as a WLC with a constant effective persistent length. This contrasts the behavior of long DNA chains that are known to exhibit variable persistent lengths depending on the ion concentration levels and force magnitudes. PMID- 16711069 TI - Regulation of vascular smooth muscle cells and mesenchymal stem cells by mechanical strain. AB - Vascular smooth muscle cells (SMCs) populate in the media of the blood vessel, and play an important role in the control of vasoactivity and the remodeling of the vessel wall. Blood vessels are constantly subjected to hemodynamic stresses, and the pulsatile nature of the blood flow results in a cyclic mechanical strain in the vessel walls. Accumulating evidence in the past two decades indicates that mechanical strain regulates vascular SMC phenotype, function and matrix remodeling. Bone marrow mesenchymal stem cell (MSC) is a potential cell source for vascular regeneration therapy, and may be used to generate SMCs to construct tissue-engineered vascular grafts for blood vessel replacements. In this review, we will focus on the effects of mechanical strain on SMCs and MSCs, e.g., cell phenotype, cell morphology, cytoskeleton organization, gene expression, signal transduction and receptor activation. We will compare the responses of SMCs and MSCs to equiaxial strain, uniaxial strain and mechanical strain in three dimensional culture. Understanding the hemodynamic regulation of SMC and MSC functions will provide a basis for the development of new vascular therapies and for the construction of tissue-engineered vascular grafts. PMID- 16711070 TI - Compressibility of arterial wall in ring-cutting experiments. AB - It is common practice in the arterial wall modeling to assume material incompressibility. This assumption is driven by the observation of the global volume preservation of the artery specimens in some mechanical loading experiments. The global volume preservation, however, does not necessarily imply the local volume preservation - incompressibility. In this work, we suggest to use the arterial ring- cutting experiments for the assessment of the local incompressibility assumption. The idea is to track the local stretches of the marked segments of the arterial ring after the stress-relieving cut. In the particular case of the rabbit thoracic artery, considered in this work, the following criteria for radial stretches come from preliminary analysis. If after the radial cut the marked segments shorten at the inner surface of the wall and lengthen at the outer surface while remaining unchanged in the middle of the wall then material is locally incompressible. If, however, the marked segments remain unchanged at the surfaces while lengthening in the middle of the wall then the material is locally compressible. Any other scenario would be an indication of the improper modeling assumptions, i.e. residual stresses are not relieved or material constants are inaccurate etc. It is believed that the proposed approach can be successfully implemented in experiments shedding new light on the arterial incompressibility issue. PMID- 16711071 TI - A mathematical model of cell reorientation in response to substrate stretching. AB - It is well documented that in response to substrate stretching adhering cells alter their orientation. Generally, the cells reorient away from the direction of the maximum substrate strain, depending upon the magnitude of the substrate strain and the state of cell contractility. Theoretical models from the literature can describe only some aspects of this phenomenon. In the present study, we developed a more comprehensive mathematical model of cell reorientation than the current models. Using the framework of theory of non-linear elasticity, we found that the problem of cell reorientation was a stability problem, with the global (Maxwell's) criterion for stability. For the case of uniaxial substrate stretching, we showed that cells would orient away from the direction of substrate strain such that the angle between the cell long axis and the direction of the substrate strain would increase with increasing magnitude of the strain. We also showed that at a given substrate strain this angle would be greater in cells having greater contractile strain. These results are consistent with experimental observations reported in the literature. PMID- 16711073 TI - [Educating, informing]. PMID- 16711072 TI - [Emergency nurses on all fronts]. PMID- 16711074 TI - [Clinical management. A always current tool]. AB - Clinical reasoning is the fundamental tool every clinician must master. This paper gives a definition of clinical reasoning process using real cases and presents some of the cognitive models underlying it (especially the script concept). We discuss the caveats of clinical reasoning process and its impact on interdisciplinary work, the use of technologies and medical teaching. PMID- 16711075 TI - [Elements of clinical care]. PMID- 16711076 TI - [Evolution from singularity to interdisciplinarism]. PMID- 16711077 TI - [Clinical research in psychiatry: what role for nurses?]. PMID- 16711079 TI - [Clinical research. Problems and organization]. PMID- 16711078 TI - [Research in clinical psychology]. PMID- 16711080 TI - [Clinical research. Empathy and therapy]. PMID- 16711081 TI - [Development of an experimental methodology: hypotheses and variables]. AB - In this methodological paper, we present the elaboration of the experimental design, central stage of the research cycle. First, we focus on the hypothesis elaboration from the research question. Then, we detail the hypothesis operationalisation by the means of the choice of variables. Finally, the experimental controls and the factorial designs are presented. PMID- 16711082 TI - [Clinical nursing research: academic medical center of maternal-child hospital St. Justine]. AB - The progression of nursing research within clinical settings is lingering. Favouring the emer gence of a research culture in these environments requires concerted action; it is effectively the only way real change can occur. It is within this perspective that an association was made between the directions of the Nursing Faculty of the Universite de Montreal, of the Research Center and Nursing Direction of the CHU Sainte-Justine (CHU). This experience is unique and deserves attention for the benefit of nurses and the advancement of the profession. This article describes the strategic plan elaborated to this end and the major accomplishments fulfilled to this day. PMID- 16711083 TI - [Suffering: representations and challenges]. AB - Research that we carried out aimed to analyzing the suffering as an object of the relation of care and common experiment of this relation. In this way, we have conducted 55 interviews with patients and nurses in an oncology unit. The results underline the central role of the relation for giving meaning to the suffering. The knowledge of the suffering representations appears as a tool for understanding the complex realities connected to the experiences of suffering, and allows to imagine more adapted evaluation methods which could be anchored on "practical knowledge". PMID- 16711084 TI - [Expected benefits of a nursing consultation on the compliance of the adolescent renal transplantation patient]. AB - Benefits expected from a nurse consultation on the therapeutic compliance of the student with a kidney transplant The therapeutic non-observance of chronic ill teenagers in general, and of teenagers who had a kidney transplant in particular, is a genuine problem of public health. At the children Hospital of Toulouse, from March 2002 to March 2003, a nurse consultation coupled with the medical consultation, was set up for the follow-up of these teenagers. The object of this research in nurse care was to appraise, at the end of a year's operation, the results of this consultation on the therapeutic observance of these teenagers and its operation within the institution. At the dawn of a deep upheaval for the profession, this study enabled us to valorise the nurse expertise combined with the medical activity for the actual benefit of children. The nurse consultation, in the heart of the news for some time, has remained in the heart of our concern for over half a century. PMID- 16711085 TI - [Clinical approach in psychiatric nursing: study of an advanced practice case]. AB - Nowadays many people suffering from severe and persistent mental disorders are cared for in ambulatory settings by multidisciplinary teams. Nurses take an important part in these teams. What is nursing practice in this type of setting? A case study was developed from a nurse's clinical practice in order to answer this question. The results suggest that these nursing services are of an advanced practice nature as defined by Hamric et al. (2000). PMID- 16711086 TI - Beyond the professional fee: basic legal issues for physicians investing in the technical side of healthcare services, part 2. AB - As reimbursement for professional services shrinks compared with physicians' educational debt and practice expenses, physicians are looking for additional sources of revenue to supplement their practice income. A natural option to consider is investment in healthcare entities or equipment in order to share in the technical fees generated by such entities or equipment. Part 1 of this article reviewed relevant legal authorities and common investment opportunities. Part 2 looks at issues that physician investors face-specifically, the choice of whether to invest in a separate entity or to expand an existing practice to include a technical service, hospital investors, and service agreements. PMID- 16711088 TI - The number one myth of asset protection: why you can lose assets in a medical malpractice lawsuit. AB - The thinking of many physicians around the country, and unfortunately their advisers as well, is that there is little risk of a physician losing personal assets in a malpractice claim, especially with typical dollars 1-3 million malpractice insurance coverage. This article dispels that myth and shows that the benefit in engaging in asset protection planning far outweighs the cost. PMID- 16711087 TI - Liability for unfounded "fear firing" under ADA: a real threat of violence, or just the postal worker metaphor? AB - Employers are sometimes confused with employee conduct that is truly a menace to the welfare of co-workers. Often, unusual behavior is perceived by others as being threatening but is not truly so. Managers must differentiate and act appropriately under the circumstances. This article outlines somes of the situations that may arise and provides guidance for managers' response within the rules set forth by the Americans with Disabilities Act (ADA). A policy of zero tolerance for disruptive acts may serve most practices well. The appropriate use of professional evaluations and employee assistance programs (EAPs) is also discussed. PMID- 16711089 TI - Minimizing medical litigation, part 2. AB - Provider-patient disputes are inevitable in the healthcare sector. Healthcare providers and regulators should recognize this and plan opportunities to enforce alternative dispute resolution (ADR) a early as possible in the care delivery process. Negotiation is often the main dispute resolution method used by local healthcare providers, failing which litigation would usually follow. The role of mediation in resolving malpractice disputes has been minimal. Healthcare providers, administrators, and regulators should therefore look toward a post event communication-cum-mediation framework as the key national strategy to resolving malpractice disputes. PMID- 16711090 TI - Simple ways to improve staff productivity. PMID- 16711091 TI - Improving medical practice outcomes by retaining clinicians. AB - Turnover of medical care providers has become so commonplace that callous disregard or weary resignation are prevailing sentiments among remaining staff members when a colleague leaves. This article analyzes reasons for turnover of caregivers and the consequences. Turnover is particularly detrimental in medical practice because it undermines learning, as well as acquisition of judgment and adaptability. Medical practice managers may be unaware of the magnitude of the hidden costs-financial, strategic, and quality-associated with turnover. Strategies are proposed to assist medical care organizations in retaining clinicians and thereby improving healthcare effectiveness and efficiency. PMID- 16711092 TI - Working with patients with physical disabilities. AB - Medical practices need to make a number of adaptations to ensure that their facilities and staff are accessible to patients and other office visitors with physical disabilities. This article describes 10 specific strategies for preparing your medical practice for patients with disabilities, both inside and outside your office. It describes minimum standards for office doorways, sidewalk inclines, ramps, reception areas, and other adaptations medical offices need to make. The article also describes specific do and don't advice for communicating with patients with physical disabilities. It suggests strategies for preparing written materials for disabled patients, for communicating verbally, and for providing healthcare education. Finally, it suggests an appropriate role for medical practice staff in the delivery of healthcare services to patients with physical disabilities. PMID- 16711093 TI - Benchmarking physician performance, part 2. AB - Part 1 of this article (January-February 2006) reviewed ways of measuring the work of physicians through methods such as data envelopment analysis (DEA) and relative value units (RVUs). These techniques provide insights into: 1. Who are the best-performing physicians? 2. Who are the underperforming physicians? 3. How can underperforming physicians improve? 4. What are the underperformers' performance targets? 5. How do you deal with full- and part-time physicians in a university setting? Part 2 compares the performance of 16 primary care physicians in the same medical specialty using DEA efficiency scores. DEA is capable of modeling multiple criteria and automatically determines the relative weights of each performance measure. This research also provides a preliminary framework for how work measurement and DEA can be used as a basis for a medical team or physician compensation system. PMID- 16711094 TI - Building an emotionally intelligent medical practice. AB - This article reviews each of the five components of emotional intelligence, describes their relevance to the management of contemporary medical practices, and provides some practical steps health professionals and practice managers can take to enhance emotional intelligence. The benefits derived from such a review will necessarily vary from person to person and practice to practice, depending on the baseline from which we begin. Nevertheless, all of us can benefit from learning more about emotional intelligence. PMID- 16711095 TI - Five things patients want. AB - Many in the healthcare industry think it's inevitable that we're heading toward consumer-directed healthcare (CDHC). For CDHC to work, it will require more of a partnership between doctors and patients than ever before. The author offers key ways doctors and their staffs can respect the time and intelligence of their partners in health--their patients. PMID- 16711096 TI - Mass vaccination clinics versus appointments. AB - Influenza vaccine is a safe, effective, and cost-effective intervention that can prevent serious disease in adults. Opinions differ as to the most effective method for delivering the vaccine to the greatest number of high-risk adults. The objective of this article is to compare immunization delivery of influenza vaccine to high-risk adults during two types of clinic visits: routine scheduled appointments versus mass clinics. Data was collected at 15 ambulatory care settings on 599 patients 50 years and over from October 23, 2001, to January 31, 2002. Immunizations given at either routine scheduled visits or at mass influenza immunization clinics were compared for costs and resource requirements (productivity), and completeness of delivery of quality visit components (efficiency). The two visit types presented significantly different strengths on key clinical functions. Routine scheduled appointments promoted more review of patient health history and more of the contact information necessary for reminder/recall and audit functions. In mass immunization clinics, patients were more likely to be vaccinated, with far less time spent in either direct services or in waiting, and it was more likely that the required vaccination information statements (VIS) would be provided. Mass vaccination clinics and routine scheduled appointments are both viable service strategies for delivering influenza vaccines. This study suggests the greatest advantage occurs when both strategies are used in a coordinated manner. PMID- 16711098 TI - Financial independence: a mighty stretch but by no means out of reach. AB - Financial independence refers to your ability to maintain your desired lifestyle without having to work. Attainment of financial independence will provide you with peace of mind, as well as many options for your future. It can allow you to switch careers and accept a lower income, continue working at a less rigorous pace, or simply enjoy an early retirement. This article covers the basics of financial planning, including questions that guide the reader to a foundation on which to build. PMID- 16711097 TI - Educate patients on billing policies to improve your bottom line. AB - In no other industry are the consumers of goods or services so unaware and uninvolved in the reimbursement for the goods or services received. The United States healthcare industry has created and allowed to perpetuate a "hands-off" compensation approach that has created a consumer mentality proving quite difficult to change. In this article, the author describes the origin of this outdated approach and suggests a restructured office and clear communication with patients as starting points to change this deep-rooted mind-set. He describes how an up-front approach to patients' financial responsibilities and proactive staff can help reduce miscommunication between provider, patient, and insurance carrier and improve your practice's cash flow. The author also suggests that the patient's financial burden will continue to increase and offers tips to maintain positive patient relationships, improve your accounts receivable management, and protect you financially. PMID- 16711099 TI - Pay for performance: its influence on the use of IT in physician organizations. AB - The introduction of information technology (IT) in physician organizations and practices is a source of great interest to physician leaders and policy makers. In this article, the authors describe what may be the nation's largest pay-for performance program, its performance metrics, and incentives for the implementation and use of IT in medical groups and independent physician associations (IPAs). Results include the increased use of electronic clinical data, point-of-care technology, and the generation of more actionable reports for quality improvement. Noteworthy are the efforts by physician organizations to enhance data collection to demonstrate improved clinical performance and earn financial incentives. PMID- 16711100 TI - Staff development on one leg: providing training with limited resources. PMID- 16711101 TI - Employer allowed to correct mistake in benefit calculation. PMID- 16711102 TI - [Safety is a dance. Training and information on prevention and safety in construction]. AB - The construction site seems a place of fragmentation and of continuous resettings of operations and plannings, attentions particular and attention to the "final product", relations and deliveries, of interests and of cures. The construction site is prepared true in different spaces and times intermittent: space and times that define unknown contexts and new relations also in presence of operations (relatively) standardized. Around these critical joints--to the good ability to recompose and to pass the deliveries (acquaintances, data, duties...)--it's played the issues of product's quality, the job and those of the emergency. A formative horizon is always in game when they open organizational experiences and of complex job that is introduced with elements of oneness, and when operations are demanded recurrent of construction of acquaintances, remittances to point of times and operations, logons of attentions, activity and various techniques. The interaction of single operators with the context, the redesign itself of the same context, render active and responsible the relation with the companions of job (of the own enterprise and of the others), they make an experience of continuous Coformation, of government of the relations and construction of comunicative pattern. To feel "dancing part" is one action and knowledge modality not only moved from cognitive and logical elements, but also emotional. It is ability to learn to trust itself and to being reliable, without to think about to hold under control the manners, the times, the outcomes of that it is gone making. PMID- 16711103 TI - [Training and information in construction in Italy]. AB - The formation and the information are at the base of the prevention of the accidents and the professional illnesses: this principle is very present in all the European directives that consider the problems regarding safety and health in the places of job, from the mother directive (take in together to other important European milestone with our legislative decree 626/94) to the building site directive and to the machine directive. Focusing the attention on the sector of the constructions, it appears immediate to think about the formation and the information addressed to the workers, that is, to those people that realize the work in first person and that could be object to accidents and job illnesses. Correctly the building site directive (and the correspondent legislative decree 494/96) involve other people of the productive process who, with their choices, their behaviors, their awareness--and therefore their formation--strongly influence the safety conditions: it deals with the coordinators in planning and in executive phase that offer, through their competence, support to the safety during the whole productive process with specific professionalisms (from the planning to the execution, from the maintenance to the demolition), with the person responsible of the firm that has to have the abilities in the job organization and in the formation of his own employees, with the buyer that, extraneous up to the 494/96 emanation to the technical-productive problem list, has to create the fittest conditions to build, to manage and maintain the work under safety conditions. PMID- 16711104 TI - [Role of the ISPESL in quality of focal point of the European Agency for health and safety at work]. AB - The building it re-enters more between the extended fields in Europe, with a advanced volume of transactions to 900 billions of euro to the year and beyond 12 million workers with reference to the single Union to 15. In the region, the implication of the medal is represented from record minimums of emergency and health on job (SSL), for one cost to cargo of the enterprises and the contributors that goes around every year around to 75 billions of euro, without to mention the uneasiness to the persons. Although important progresses caught up in the course of the time, faces to improve standard of SSL of the building industry, thanks to one more effective cooperation between employers, workers and others you leave interested, the job to make it is still very many. This year, European week for the emergency and the health on the job, dedicated to the field of the building, it is proposed to support every interested part of the field in the putting to fire of it upgrades them improvements in order to create job atmospheres surer, you heal and productive. The campaign, directed from the Agency for emergency and the health on the job, in its turn financed from the European Union, it will reunite a wide review of activity times make more aware on the main ones problems and the respective solutions. The present informative package has been created in order to supply information it generates them inherent to such problematic let alone the elements necessary in order to participate to the campaign. PMID- 16711105 TI - [Role of the INAIL]. AB - This report has to describe the activity of INAIL for improve information and training in the field of occupational health and safety. The discussion start from accident and professional illness data and gives evidence about social and economic cost of this phenomenon. INAIL want work together with other institution, social body, professional association for promote the integration of occupational health and safety in businesses management. The principal target are SMEs - Small and Medium size Enterprises. Information and training must follow this approach and sustain it. Information and training have to develop knowledge, competence and awareness like value for the organisations. This report describe models and tools for information and training which INAIL's improving even for the building sector. PMID- 16711106 TI - [Training and information for coordinators]. AB - The formation and the information, announced by the mother directive and recepiti with the legislative decree 626/94, they are essential actions that the employer has to put the prevention of the accidents and the professional illnesses towards the workers into effect. They are fundamental elements for the safety and the health on the places of job, the formation and the information they bring the inherent dangers to the knowledge of the worker in the duty that has to develop, in the useful technologies to the productive trial and the place where the productive trial is in action. Transferring to the sector of the constructions the formation and the information that the employer has to guarantee is immediately thought about the workers that realize the work and that youlthey can result then object of accidents and job illnesses. The directive yards 92/57CEE and subsequently the D.Lgs 494/96 involve the subjects that with their decisions, their actions, consequent to evaluations and choices operational respect the work to be realized they condition the safety. The subjects that they compile the safety plan and coordination with their formation and they manage the safety in the executive phase are the coordinators. Necessary is the formation of subjects that with theirs to act and their choices could condition the safety of other interactive subjects in the phases of job that characterize the work, therefore it is fundamental to individualize the necessary components to their formation. PMID- 16711108 TI - [Training in health and safety: the strategy of the ISPESL]. AB - Activities on OSH training and information carried out by ISPESL--Documentation, Information and Training Department--have been greatly improved by the implementation of the European Directives into the Italian Regulations. Through the planning of a large number of researches, realized by the above mentioned Department in cooperation with Universities, Regions and Social Partners, a correct and efficient training policy has been set up, supported by the realization of innovative and suitable didactic tools. Great attention has been devoted to the improvement of quality of the training process, entirely analyzed in order to transform OSH training into a real investment to companies and workers. PMID- 16711107 TI - [Training and information for competent physicians]. AB - The rule in force and the Guide-Lines concerning the topic point out specific duties for the Occupational Physician; the most important is certainly health survey, which is organized in several formal medical actions; another Occupational Physician's relevant activity is the "collaboration to the risk assessment". Because of the complexity of the construction building industry and because of the central role of the Occupational Physician in health care, the problems which he has to face are two: firstly he has to improve the knowledge about specific risks of the construction building industry; secondly he must adapt his specialistic knowledge, including the methodology acquired during the school training, to the specific organization conditions and risk of the field. PMID- 16711109 TI - [Training courses for coordinators in matters of safety and health]. AB - Bill 528/99 increased the number of technicians who can carry out the tasks of the Safety Coordinators after having attended a specific course. In order to do this and to give precise indications regarding the contents of these 494 Courses, it was decided to issue a special Decree to deal specifically with this matter. In spite of the fact that the deadline agreed on for the issue of this Decree expired almost four years ago, no regulatory act has yet been passed. Although in no way wanting to act as a substitute for those delegated by the Legislator, this contribution will put forward a plan for the 494 Course and will also define a didactic methodology and different contents concerning the qualification and specialization of the technicians to be trained. PMID- 16711110 TI - [Training quality and quality training]. AB - There is a growing need for all organizations to carry out in-house training based on concrete strategies to be able to face the risks involved in the working context. The area that is most in need of this training is the building sector as it has always been penalized by the poor qualification and cultural levels of its operators, which are among the lowest in the working world. The need to determine in-house training which meets this requirement calls for a particularly effective practical and methodological approach. To achieve this aim, not only is it important that the training programme and the trainers fulfil the requirements of the Legislator, but they must also aim at both improving the technical quality and increasing the qualifications and cultural growth of the human resources. The purpose of this contribution is to advance a correct methodological approach aimed at training quality and quality training. PMID- 16711111 TI - [Safety measures to prevent falls are introduced into the training course for people in charge of planning]. AB - For to pursue and to stimulate adequate shapes of prevention of the risk of falls from the high, the ASL of the province of Bergamo has approved of the moderniization of the R.L.I. that it introduces the obligation of the predisposition of structural elements that they favor the successive one rests in work and 1 use of DPI. It has been activated, therefore, a Work group composed from doctors and technicians of the Sanitary Company and graduating of the Faculty of Building Engineering of Bergamo, with the obbiettivo of to create means of support for the been involved figures of system. Us one is prefixed this obbiettivo to useful the aim to predispose and to put into effect course to to answer to the needs formed you of the professional figures of system and not in matter of risk of fall from the high. PMID- 16711112 TI - [Intervention methodology for training and information for workers of the building sector]. AB - Regarding the seriousness of work-accident in Gorizia district, various organizations as ASL 2 "Isontina" (local health agency), INAIL of Friuli Venezia Giulia (National Institute for occupational accident insurance), trade unions and trade associations created a organization called "Observatory for Working accidents and Professional Illness Prevention". The aim of this association is the promotion of safety in working environment. Diffusing importance of safety in building trade was the first projects of Observatory. So, Observatory carried an initiative to make more aware the workers. Than, it organized training courses for building workers. Moreover, the construction of an informative pamphlet on risks in building trade was made. For some experimental investigation, Observatory works with Department of Psychology, University of Trieste. PMID- 16711114 TI - [How young people picture construction]. AB - The aim of this research is to analyze qualitatively and quantitatively the representation of housebuilding filtered by young people. The main instruments used for the qualitative phase were not-structured interviews to adults placed in building field and to young people between 14 and 25 years who are entering this world. So, the results show two visions of housebuilding that, although they largely coincide, from some points of view they diverge. PMID- 16711113 TI - [Risk perception of the building population]. AB - The development of a new measure to investigate risk perception in afield setting as building was the aim of this work. Workers' cognitive representation of risk and relation among ris perception, risk-acceptance and risk taking was the goals of the measure. The methodological approach for the measure development was the psychometric paradigm (Fischhoff, Slovic, Lichtenstein, Read e Combs, 1978). A first data collection in field context demonstrates measure's validity and reliability. PMID- 16711115 TI - [Educational health programs in the construction industry of the Apulia Region, southern Italy]. AB - 31,000 building firms have been registered in the Apulia Region of Southern Italy, and 13.5% are concentrated in the area of the Local Health Bari/3. Especially small and middle-size firms (90% of total number) are assisted with information programs through dedicated local office. Educational and medical audit advanced programs carried out among occupational health professionals are described. A 'pilot project' for the regional health surveillance program in the construction industry is presented. PMID- 16711116 TI - [On the job training at construction sites of the hospital corporation of Careggi (Florence)]. AB - The "Careggi Safety" project has like objective to guarantee the safety execution of Careggi Hospital (Florence) restoration and development works, thanks to an in itinere training program into building sites and through tutor figure using. The project aim is to overcomes traditional indoor training limits, not effective in complex and dynamics reality like as building sites, constrained by contracts deadline and high labour turnover (subcontracts) inside carry out process. Solutions chose are: (a) a training projected in itinere, following site works evolution and safety and coordination plan, and through a constant agreement between customer and operative enterprises; (b) a building site's tutor, standing beside workers during realization phases, contributing to form on respective safety carry out job. In to "Careggi Safety" project training has been chose as preliminary and obligatory condition for labours admission and control into the building site. PMID- 16711117 TI - [Integration and safety at the workplace]. AB - Creation and experimentation of one specifically destined module of formation to emigrated workers buildings employ to you in companies craftsmen. It realizes 4 to you course for to face the topics of the emergency and prevention in the yards (obligation formative D. Lgs. 626/94), supplying to contempo the instruments in order to facilitate the acquaintance of Italian language and integration in the society and the atmosphere of job. PMID- 16711118 TI - [Training individuals of the construction sector in the province of Bergamo: a shared path division and an operative proposal]. AB - With this presentation it will be introduced the experience of the work group native of Bergamo who is taken care to plan the formation course for the professional figures of system in building. The group is formed from the greater agencies present on the territory, their work is to ensure the protection of the health and the emergency of the workers in the field of the constructions. The objective is to use the formation course in order to diffuse one culture of the emergency. In the specific one they will come to describe the various planning phases of the course of formation for employers that mean to elect himself Responsible of the Service of Prevention and Protection (RSPP). PMID- 16711119 TI - [Management of the new first aid service at enterprises and construction sites]. AB - First aid at work organization and management represents a complex and critical aspect of the manifold problems of hygiene and security at work; nevertheless, even in relatively well organized productions, these themes are often neglected, if not completely ignored. In this work the authors analize the laws which regulate the correct first aid at work organization and management and an approach to the problem based on the preliminary risk assessment is suggested. PMID- 16711120 TI - [Ministerial decree 388/03: application problems in the construction sector]. AB - The introduction of D.M. 388/2003 about first aid procedures to be carried out in the place of work has brought specific questions on how to enforce them in the building industry field. The aim of the text is to deal, though not in a specific way, with the relationship between the enterprises and the ASL [Health Authority] in order to prepare First Aid Plans. More specifically, it deals with the duties of the Employer, of the safety supervisor, of the doctor in charge and of all the people responsible for helping. There are also strategies on how to plan first aid measures in the building industry and suggestions to supplement materials in the First Aid kit. PMID- 16711121 TI - [Role of the Services for Prevention Safety at the Working Setting (SPSAL) in the application of the Ministerial Decree of July 15, 2003, No. 388 (regulation on the organization of first aid in the construction sector]. AB - Article 15 of the D.Lgs. 626/94 and D.M. 388/2003, performance regulations, have codified of the new modalities of management and organization of the Service of Ready business Aid. I previewed in the new approach, entrusted main tasks to the employer, they are of organizational and formative type. PMID- 16711122 TI - [Role of the Department of Emergency in the application of the MD 388/03: aid, treatment and prevention]. AB - The aid happens behind call of national the only number for the sanitary emergency: 118. The times of Aid you flavored yourself and indicates to you from the Institutions, regarding the moment of call, previews the arrival of a aid characterized within 8 minuteren for the area to high inhabited density and within 20 minuteren for the areas to lowland density. The Means breakup of Aid near yards with particular infortunistico risk, is possible, and case for case must be estimated: the demand must be placed to the responsible of It centers them Operating 118. Legislator previews that the reported formation, to the practical ones of ready aid, of the attache's to the emergency of the enterprises it is made from the where possible doctor 'in collaboration with the system of emergency-urgency of the sanitary service nazionale'. Also the demands for formation for specific risks must to be agreed with the responsibles of the Structures of the Emergency of the SSN in particular the Directors of the Structure of Ready Aid, of Center them Operating 118, or of the Department of Emergenza-Urgenza. PMID- 16711123 TI - [Health emergencies and first aid at construction sites in the Tuscany Region. The high speed and varying free way of Valico]. AB - From 1996 the territory of Tuscany region and in particular that one of ASL 10 of Florence, has been interested from the realization of great works: the railway line to High Speed and varying of Valico of the A1 freeway between Florence and Bologna. ASL 10 has stipulated specific economic agreements with the been involved companies in the realization of the plans for a plan finalized to the prevention of the industrial accidents and to limit of the gravity with timely aids it adapts to you also in gallery, to guarantee to the workers the same performances sanitary of the city residents, to guarantee and to improve the quality of existing sanitary services on the territories in spite of the increase of the population weighing on the consequent territory to the takeover of the workers it engages to you in the realization of the great works. This plan has been realized with the creation of integrated aid system a to strengthen the territorial sanitary emergency, I extend the access to the base medicine to the workers and has not been able itself to answer to 93% of the question sanitary to the inside of the same yards with the infirmaries of yard without to resort to external structures. PMID- 16711124 TI - [First aid training in collaboration with CPT and CPTA of Bergamo]. AB - In this work it's briefly described the experience in planning and realizing of first aid at work courses for workers of the building trade, realized by the UOOML in collaboration with CPT and CPTA of Bergamo. In particular this is a report of the activity of planning and realization of courses in the months from February till September 2004. PMID- 16711125 TI - [Assessment of the efficiency of the first aid course among construction companies in the Province of Bergamo]. AB - The aim of this project is to value, through a specific questionnaire, the effectiveness and utility of the professional training among the construction workers responsible of the First Aid. 48 construction firms, in Province of Bergamo, were examined and 100 first aid responsible were tested through to the questionnaire. The 21% of the subjects performed a practical first aid intervention. The 84% considers the professional training courses to be globally satisfactory, and one third underlines the necessity of an annual refresher course. This study tends to confirm the validity of this program of professional training, particularly in the construction trade, that is characterized by a high accidents rate. PMID- 16711126 TI - [The formation and the information of the prevention and safety in construction]. AB - The relation takes in consideration puts into effect it them adequacy of the information, the formation and of the training in building. It distinguishes between information, formation and training obbligatori and opportune. It indicates the necessity that the contents of the information and the formation they are: specific, punctual and it adapts to you; adaptable to the various situations, acceptable, effectively feasible; with you specify objects to you to catch up; sostenibili in terms of cost. And above all that they work for objects to it to you that us had been placed. The relation indicates which strategies an employer can follow in order to catch up adequate and a reasonable one level of informazione/formazione and which it is, today, the situation of our enterprises buildings, analyzing like the evolution of the labor market and the use of sub the contracts goes, in consisting way, to modify the situation pre-esistente. It analyzes many of the situations industrial accident researches for which the information, the formation and the training they introduce unsatisfactory. It tries to give answers to following interrogated to you opened: Which duration and which contents must have theformation? To who it goes turned? Who must be supplied? To such aim it distinguishes between: course very defined (from specific norms) for contained and duration, Iter you drained to us, course rendered obbligatori or opportune from the enforced norms, but does not codify to you. It characterizes in the employers of the small enterprises, in the attache's it specializes to you and generic and in the futures professionals of field during the scholastic formation the subjects give to privilege in the formation. It reconstructs the offer of formation, information and information available today and which addresses they can be develops to you. PMID- 16711127 TI - [Prevalence of risk factors and prevention of coronary disease in a population of construction workers of the Province of Bergamo]. AB - Authors describe the prevalence of cardiovascular risk factors among 1.348 construction workers, enrolled in a five years lasted health surveillance program. Our results confirm both the high prevalence of cardiovascular risk factors and the need of further examinations other the clinical, laboratory, electrocardiogram and chest X, in order to point out a correct diagnosis and evaluate the fitness to job. Furthermore, we underline the usefulness of training in health education for the prevention of cardiovascular disease, considering the low extent to which individuals perceive their own risk. PMID- 16711128 TI - [Prevention in the building sites of the District of Sondrio]. AB - Building is one of the most important activities inside the local productive reality; the PSAL Service has therefore had to face the sector intervention prearranging a specific methodology, which would aim at involving all the people in charge of roles and authority in this field, finding out a way that, starting from the epidemiological evaluation of accidents, would become positive making the shared preventive standards, into which the watch activity, the training of the employers, workers and professionals, the effective involvement of public contract giving stations would be concentrated, well-grounded. The first results of such an important engagement are ripening: there are still some mortal accidents but as far as the other cases are concerned we point out that all the points on which the watch activity has been concentrated (fall of people and of gravel from high points, contact with working elements of the installations, etc.) are constantly decreasing. The estimation of the risk priority has been carried out under mandate from the Procura della Repubblica resulting in the establishment of a table in which all the main working conditions with a high risk of people and gravel fall from high points, transformed into a special Criminal Investigation Department Directive on the cases on which to concentrate the intervention and in the presence of which to distrain upon/to operate sequestration. The involvement of various people in charge of prevention in the yard has occurred during the District Commission of Coordination ex art. 27 DLgs626/94 (ex article 27 Law626/94) which, thanks to a proper Building Working Team, has divided the above mentioned Directive of the Procura della Repubblica into guidelines addressed to each component and consequently into the training of the categories involved. Therefore, a guideline for the building constructors has been produced in 2000; such guideline has been adapted to professionals in 2001; a series of enterprises concerning capillary agreement with the Local administration in order to point out the essential problems regarding prevention inside the administrative procedures concerning the management of the public contracts in the field of security are being carried out since 2002. PMID- 16711129 TI - [Safety and training. Collaboration experience among degree courses in Sciences of Education and PSAL Service]. AB - This presentation shows which could be the possible links between the Bergamo University and its area. In particular it treats a cooperation between the "PSAL service of Bergamo ASL" and the degree of Scienze dell'Educazione in regard to safety on work in field of building construction. After a short introduction which shows the essential features of training, we show pragmatically two experiences achieved in different moment, by two students of Scienze dell'Educazione at PSAL service. PMID- 16711130 TI - [Quality assurance to efficacy-effectiveness measuring in occupational medicine]. AB - The occupational medicine was asked to play an important role in quality assurance procedures as demonstrated by the implementation of main principles of prevention in ISO EMAS and SA schemes. Peculiar condition appeared to be the promotion of quality in occupational physician activities as, for example, provided in these last three years the Italian Society of Occupational Medicine and Industrial Hygiene (SIMLII) by its excellence accreditation programme. The next step will be the identification and application of adequate indicators of efficacy-effectiveness of occupational medicine practices to demonstrate need and role of our discipline in guaranteeing and improving health in working places. To these aims has been addressed the meeting held in Bergamo and organized by the Lombard section of SIMLII in December 2005 whose main contributions are published in this GIMLE special issue. PMID- 16711131 TI - [Research and formal demonstration of evidence of efficiency in occupational medicine]. AB - There is an increasing pressure on occupational health professionals to ensure that their practice is based on a quality standard and an evidence of appropriateness. The efficacy-effectiveness of their activity strictly depends on their intellectual and professional integrity, their technical and legal competence, but also on the quality of the relations with the company organisation they are able to establish. We asked to some occupational physician when they considered their interventions in the workplace to be efficacious. Then, accordingto our epexrience, we propose some effectiveness indicators, particular concerning health surveillance and health educttaon, that should be evaluated to verify the health professional agccvity agreement to quality standards. PMID- 16711132 TI - [Occupational epidemiology and efficiency evidence]. AB - Occupational medicine is undergoing major changes and the question of "evidence" seems essential at this turning point. We need to update knowledge, adjust methods and verify the efficacy of interventions. If this is not done, the discipline might be reduced to just passive application of legislation with a consequent reduction of occupational medicine's professional and scientific content. Procuring evidence is objectively more difficult in the field of prevention than in the clinical field; however, those difficulties should be addressed and not simply handled in a trite way. In occupational medicine there is special opportunity to combine a "population" approach (tending to reduce exposure of the entire population) with a "high risk" approach where particularly susceptible subjects are identified and treated with targeted interventions. The contribution of epidemiology to the goal of underpinning a renewed occupational health practice with proves of evidence, mainly rests in suggesting methods able to (1) guarantee the validity of the observation of relationships between exposures and effects; (2) put together observations and studies independently conducted on the same issue and produce a combined quantitative evaluation; (3) set up ad hoc studies for evaluating prevention measures in progress. Health surveillance programs represent a particularly fertile soil for evaluation research and evidence accrual, however they are not sufficiently taken into consideration for this purpose, and relevant studies are few. A renewed cooperation involving occupational medicine and health centres is necessary to achieve that goal. PMID- 16711133 TI - [Evidence-based medical surveillance in occupational health]. AB - One of the major issues in Public Health is the effective allocation of resources to appropriate and efficient interventions. Medical surveillance is a part of the complex management of prevention in the Occupational Health and Safety field. The aim of the study is to present the efficacy and effectiveness of medical surveillance through an overview of articles and reviews (only one systematic review) found on scientific electronic databases (PubMed and Embase). Although the literature reports a lack of data, two main themes were selected and discussed: the quality of preventive actions and the cost-effectiveness of medical surveillance. PMID- 16711134 TI - [Good practices and quality in the practice of occupational physicians]. AB - This contribution describes practice and tools which allow occupational physicians the realization of theirs tasks not only respecting law and ethical codes, but also in view of a continuous improvement of their activities and customers (companies) and consumers (workers) care. PMID- 16711135 TI - [Efficacy in occupational medicine: proposals of the regional sections of SIMLII of Lombardy and Emilia Romagna]. AB - The efficacy in Occupational Medicine is based on the degree of attainment of workers health and safety care. The Occupational Physician activity, attending on business parts, gives a contribution to the mission of the firm leading to the workers health/safety. The Occupational Physician actions are conditioned by workers features and by the other members of prevention system. It is necessary to verify the efficacy, the efficiency and the adequacy of the Occupational Physician activity finding goals and specific indicators. PMID- 16711136 TI - [Evidence based occupational health: from theory to practice]. AB - BACKGROUND: Health care organizations are increasingly aware of the need to apply quality assurance principles to serve their mission and there is an increasing pressure on health professionals to ensure that the practice be based on evidence of appropriateness. Therefore, medical specialists of different disciplines, including occupational health, are presently required to shift from habitual practices to evidence-based practices. OBJECTIVE AND METHODS: This paper aims at illustrating the opportunity offered by the Evidence Based Medicine (EBM), which can be used in the decision making process of the occupational physician according to the Evidence Based Occupational Health (EBOH) paradigm. The EBOH approach consists of 4 steps (i) the formulation of the problem according to the PICO model, (ii) the search for scientific external evidence based on scientific papers and documents available in different database, (iv) the critical appraisal of such evidence, (v) the application of the evidence in the decision making process. RESULTS AND CONCLUSIONS: In spite of some barriers, such as time constraint, which could prevent looking for searchable information, the evidence based decision-making process should be based on the evidence provided by major resources to answer the question emerging during the practice. Acquiring the skill for information managing facilitates searching appropriate solutions to problems usually met during the professional practice and the adoption of behaviours which will improve the practice. PMID- 16711137 TI - [Efficacy of the activities of the Prevention and Safety Services at the Occupational Setting of the Lombardy Region]. AB - The authors describe the programme of the Regione Lombardia about efficacy of the Public Occupational Health Services. The Regional law 12/03 abrogated the visits for under-age workers not exposed to risks. It is established methods to value efficacy considering the number of workers protected by the risk through prevention activity. PMID- 16711138 TI - [Clinical and risk records assessment]. AB - In the collaboration between the Hospital Occupational Health Department and Public Occupational Service of Bergamo, it has been elaborated a plan finalised to the verification of the quality and of the appropriateness in the compilation of the Clinical and Risk Record in the firm. The study has been lead by evaluation of 409 clinical and risk records (containing the first visit and, in 277 records, a periodic visit) collected from different productive areas (wood, building, health, agriculture). The analysis of the clinical and risk records have shown many critical aspects in their compilation, for this reason it is necessary to improve vigilance and attention from Public Occupational Service, Hospital Occupational Health Department and the Scientific Associations. PMID- 16711139 TI - [Health surveillance in noise exposure]. AB - The Occupational Health Department of Bergamo Ospedali Riuniti and Occupational Health Public Services set up a project in order to check the health surveillance quality in noise exposed workers. 887 hearing threshold tests, performed between 1998 and 2002, were examined; the audiograms concerned 304 workers exposed to industrial noise higher than 85dB(A), employed in 22 textile, mechanical, wood, stone, building and paper industries. After examining the 887 audiograms we noticed that the health surveillance of noise exposed workers up to now, 14 years after the issuing of the Legislative Decree 277/91, reveals many critical situations, showing an inadequate accuracy and possibility of interpretation of the audiometric tests. PMID- 16711140 TI - [Study of the health surveillance activities of the C.P.T. of Bergamo]. AB - A "customer satisfaction" research about the approving of the sanitary watching activities provided by the "Comitato paritetico territoriale" of Bergamo and the Department of Occupational Medicine of "Ospedali Riuniti" in Bergamo, comprehensive of the diferent points of view by the different subjects involved in the process, in order to estimate and improve the facilities provided. PMID- 16711141 TI - [Efficacy of periodic health surveillance of construction workers visited at the CPT of Bergamo]. AB - The aim of this work is to expose the results obtained with the application of a model about efficacy in occupational medicine proposed by ALMLII. In particular we discuss the validity of some indicators of efficacy used in a large group of construction workers during health surveillance, in order to verify effectiveness and appropriateness of an occupational medicine service, as a part of the whole prevention and safety system. PMID- 16711142 TI - [Quality and efficacy in occupational medicine: relationship between quality, efficacy, and surveillance -- Experience of public and private facilities in Emilia Romagna]. AB - Italian legislation concerning health and safety at work establishes minimal requirements, according to European regulations. On the other side it's compulsory for Employers to set themselves at a level "above minimal requirements" according only to technicalfeasibility. It's a duty of Control Bodies (in Italy National Health Service's Work Environment Prevention Services) to verify the respect of law's preview minimal requirements. Quality standards and application guidelines, on the contrary, require collaboration and consensus among all actors involved in the implementation of health and safety managing systems on workplaces: Control Bodies, Professional Officers (Occupational Physicians, Safety Consultants), public and private Consultant Companies working on the field of health and safety at work. In this document some Emilia Romagna Region Control Bodies guidelines are discussed, and two experiences concerning public and private Consultant Companies are presented. PMID- 16711143 TI - [Experience of the Parmesan Association of Occupational Physicians]. AB - The paper presents the experience of the Parmesan Association of Occupational Physicians, an important opportunity to establish a dialogue among physicians variously engaged in the promotion of workers' health. PMID- 16711145 TI - [From a wide gap to reciprocal enrichment: a wager for the Swiss Society of Allergy and Immunology]. PMID- 16711144 TI - [Efficacy evidence in occupational medicine]. AB - The author shows an overview of the most relevant features that characterize the development of the so-called "Evidence Based Occupational Health" (EBOH). After a short introduction dealing with the origin of the movement towards an evidence based medicine and health services, attributed to Donabedian and Vuori for WHO during the '80s on one hand, and, of course, fuelled by the most comprehensive Cochrane Collaboration, the most recent developments of EBOH both in Europe and in our country are discussed. PMID- 16711146 TI - [Lake Geneva Region guidelines on management of adult asthma]. AB - Asthma is a major cause of chronic morbidity throughout the world. In Switzerland, 6.9% of the adult population is suffering from asthma. The standards of treatment are unfortunately not met in most western countries, as well as in Switzerland. We put forward a complete guideline on management of adult asthma, inspired from GINA and BTS guidelines, and adapted to the specific needs of general practitioners working in french part of Switzerland. This guideline reflects a consensus between allergy, lung and emergency specialists, working in the 2 university hospitals of the Lake Geneva Region (HUG and CHUV). PMID- 16711147 TI - [Immunologic monitoring: what's new?]. AB - The monitoring of the immune response is not very sensitive. In addition, assays that could allow the quantification of the immune response are missing or not performed by most of routine laboratory. Coming from the research, new approaches and new technologies have revolutionized the monitoring of the immune response. Several of these new approaches are part of the routine to monitor the immune response. Some others, still in development, will also be part of the new assays propose by the laboratories to increase the specificity and the quantification of the immune function. PMID- 16711148 TI - [Specific interferon-gamma assays: a modern tool for tuberculosis diagnosis]. AB - Compared with the tuberculin skin test, the antigen-specific interferon-gamma assays, using a combination of two antigens ESAT-6 and CFP-10, has higher specificity for the diagnosis of latent tuberculosis, better correlation with exposure to M. tuberculosis, no cross-reactivity due to BCG vaccination and less towards nontuberculous mycobacterial infection. Fewer false positive results in uninfected persons avoid the costs of unnecessary therapy and its possible side effects. In low endemic areas, interferon-y assays are useful in addition of diagnostic algorithm for individuals with suspected tuberculosis. Further studies are required to evaluate the utility of the interferon-gamma assays in specialised subgroups of patients (immunocompromised, young children, patients with extrapulmonary disease,...) and as a marker of disease activity. PMID- 16711149 TI - [Diagnosis and follow-up of vasculitis: usefulness of imaging]. AB - Vasculitides are due to inflammation of the vessel wall. There is a definite advantage in visualizing the inflammatory process within blood vessels without resorting to invasive procedures. A variety of non-invasive imaging techniques is now becoming available to investigate patients with vasculitis. These include ultrasonography, MRI coupled to angiographic sequences, PET, single photon emission computed tomography (SPECT). Their role is being evaluated and their characteristics exploited to address issues specific to each vasculitis. Thus, cardiac IRM should be considered in patients with ANCA-negative Churg-Strauss syndrome. Moreover, PET could be useful from investigate a subgroup of patients suffering from giant-cell arteritis. However, to validate these techniques, prospective studies in large cohorts of patients are needed. PMID- 16711150 TI - [Pathogenic mechanisms in systemic sclerosis and their therapeutical consequences. Part 1: pathogenesis]. AB - Systemic sclerosis I(cleroderma) is a connective tissue disease caracterized by an aberrant immune activation, a vasculopathy and a fibrosis of skin and multiple internal organs (lung, kidneys, gut, among others). At present no unifying pathogenetic hypothesis exists to explain all aspects of this disease. The current hypothesis is that in patients with a favourable genetic background, certain environmental factors could produce alterations of cellular and humoral immunity and alterations of the microcirculation resulting in excessive fibrosis. A crucial component in systemic sclerosis pathogenesis is the persistent and unregulated activation of genes encoding the various extracellular matrix proteins. This is in correlation with different cytokines and growth factors produced mainly by T lymphocytes. PMID- 16711151 TI - [Pathogenic mechanisms in systemic sclerosis and their therapeutical consequences. Part 2: treatment]. AB - Systemic sclerosis (scleroderma) is considered as the most severe connective tissue disease. It is characterized by an abnormal immune activation, a vasculopathy and a fibrosis of the skin and of multiple internal organs. Numerous progress in the understanding of the pathogenesis with identification of key molecules have permit to introduce novel treatments that improve the management of some aspects of the disease. ACE inhibitors are effective in resolving renal crisis. Cyclophosphamide is useful for treatment of fibrosing alveolitis. Prostaglandins, endothelin receptor antagonists and phosphodiesterase type 5 inhibitors permit to improve the treatment of the vascular complications (digital ulcerations, pulmonary arterial hypertension) of scleroderma. PMID- 16711152 TI - [Somatoform disorders: diagnosis and treatment]. AB - Somatoform disorders include several diagnoses, the treatment of which concerns somaticians as well as psychiatrists. This paper defines those diagnosis features, differential diagnosis, epidemiological aspects and etiology hypothesis. Regarding treatments, emphasis is stressed upon, the high level of collaboration between somaticians and psychiatrists, regarding those long term follow-up, well known for potential high-costs in terms of individual suffering and financial terms. PMID- 16711154 TI - [Managing the dead in pandemics]. PMID- 16711153 TI - [Blisters unlike others]. PMID- 16711156 TI - [On omega 3's and the new eugenics]. PMID- 16711157 TI - [The first human neobladders produced by tissue engineering]. PMID- 16711158 TI - [Acknowledgements]. PMID- 16711159 TI - Extracardiac tissues and the epigenetic control of myocardial development in vertebrate embryos. AB - During the past few years, research on the developing cardiovascular system has given new insights into the origin and development of the myocardium in vertebrate embryos. In the present paper, a review is given on our current knowledge about two aspects of myocardial development that have been found to depend on signals from extracardiac tissues. These two aspects are, firstly, the development of the so-called heart-forming fields and, secondly, the morphogenesis of the outer compact layer of the myocardial wall. PMID- 16711160 TI - Bridging the gap between anatomy and molecular genetics for an improved understanding of congenital heart disease. AB - Birth defects are the leading cause of infant mortality and malformations in congenital heart disease (CHD) are among the most prevalent and fatal of all birth defects. Yet the molecular mechanisms leading to CHD are complex and the causes of the cardiac malformations observed in humans are still unclear. In recent years, the pivotal role of certain transcription factors in heart development has been demonstrated, and gene targeting of cardiac-specific transcription factor genes in animal models has provided valuable insights into heart anomalies. Nonetheless results in these models can be species specific, and in humans, germline mutations in transcription factor genes can only account for some cases of CHD. Furthermore, most patients do not have family history of CHD. There is, therefore, a need for a better understanding of the mechanisms in both normal cardiac development and the formation of malformations. The combining of expertise in cardiac anatomy, pathology, and molecular genetics is essential to adequately comprehend developmental abnormalities associated with CHD. To help elucidate genetic alterations in affected tissues of malformed hearts, we carried out genetic analysis of cardiac-specific transcription factor genes from the Leipzig collection of formalin-fixed malformed hearts. Working with this morphologically well-characterized archival material not only provided valuable genetic information associated with disease, but enabled us to put forward a hypothesis of somatic mutations as a novel molecular cause of CHD. Knowledge of cause and disease mechanism may allow for intervention that could modify the degree of cardiac malformations or development of new approaches for prevention of CHD. PMID- 16711161 TI - Histomorphology and ultrastructure of pancreatic islet tissue during in vivo maturation of rat pancreas. AB - In this study, we have investigated the structural and ultrastructural features of pancreatic islet tissue during rat postnatal development. For this purpose, we used neonatal (1-2 days old), young (21 days old) and adult (3-4 months old) rats. From a functional point of view, neonatal islet tissue displayed a relatively poor insulin secretory response to glucose stimulation in comparison with the adult ones. Histological analysis showed that neonatal islet cells display a less organized morphology in comparison with the young and adult ones, characterized by a less defined form and the presence of ductal structures within or nearby the islet. Regarding the islet cytoarchitecture, no differences were observed among all animal groups studied. B-cells were always typically detected within the islet core while A-cells occupied the islet periphery area. No marked differences were found during postnatal animal development regarding the ultrastructural aspect of the endocrine cells and their secretory granules. Nevertheless, quantitative analysis showed a lower B-cell/non-B-cell ratio, a higher association with ducts and an increased immunoreaction for proliferating cell nuclear antigen (PCNA) in neonatal islets as compared to young and adults. In conclusion, the acquisition of an adult pattern of insulin secretion may require an appropriate histoarchitecture and B-cell/non-B-cell proportion that may affect crucial regulatory events such as the paracrine and/or the cell-cell interaction or communication within the islet. PMID- 16711162 TI - Arterial calcification in mice after freeze-thaw injury. AB - Vascular calcification is highly correlated with atherosclerosis and cardiovascular disease and is a significant predictor of cardiovascular morbidity and mortality. Studies in mice indicate a genetic contribution to this dystrophic extra osseous calcification. We sought to elaborate a method to induce dystrophic arterial calcification in mice and further examine the pathogenetical mechanisms involved in the phenotype. We established a method of freeze-thaw injury of the infrarenal aorta producing a limited tissue necrosis and histologically investigated the occurrence of dystrophic calcification within the aortic wall 1, 3 and 7 days after injury in C57BL/6 (a mouse strain shown to be resistant to dystrophic cardiac calcification after injury) and C3H/He (susceptible to dystrophic cardiac calcification). C57BL/6 mice exhibited no dystrophic calcification at all within the vessel wall upon injury of the infrarenal aorta (0/5 mice 1 day after injury and 0/10 animals 7 days after injury). By contrast C3H/He mice displayed a remarkable extent of calcification mainly present within the media of the infrarenal aorta which was evident as early as 24 h (three out of five animals 1 day after injury) and reached its maximum extent 7 days after injury (10 out of 10 animals at the seventh postoperative day, p<0.001 compared to C57BL/6 mice). Upon immuno-histological analysis calcification was accompanied by the occurrence of certain bone-matrix associated proteins. Osteopontin and Bone Morphogenetic Protein 2/4 expression was detected co-localized with the calcified lesions. Our results demonstrate that freeze-thaw injury of the infrarenal aorta is a sufficient method to induce dystrophic arterial calcification in mice. We present evidence that the occurrence of arterial calcification in C3H/He mice seems to be actively regulated by certain bone matrix associated proteins. PMID- 16711163 TI - Morphometrical study of arteries and veins in the human sheet-like muscles (pectoralis major, latissimus dorsi, gluteus maximus and trapezius) with special reference to a paradoxical venous merging pattern of the trapezius. AB - The trapezius is one of the critical targets of physical therapy for shoulder functional disorder because this muscle plays a great role in fixation and elevation of the shoulder. Trapezius (n = 63), latissimus dorsi (n = 5), pectoralis major (n = 7) and gluteus maximus (n = 7) muscles were obtained from 43 donated cadavers, and vascular morphologies were compared. An artery without a concomitant vein was found in 35 of 63 trapezius muscles. Moreover, twice as many venous merging sites as arterial branching sites were present in the trapezius. Notably, peripheral or distal venous tributaries were larger in caliber than the proximally located vein in 6.2% of all 404 venous merging sites in the trapezius. Moreover, similar paradoxical venous merging, in which tributaries were relatively thicker than drainage capacity of the venous trunk after merging according to Poisueille's law (peripheral or distal thickness >0.85 x proximal thickness), was observed in 39.1% (158 of 404 merging sites) in the trapezius, compared to 8-9% in the other 3 muscles. Given this vascular morphology, trapezius muscle seems likely to frequently display venous blood retention. Interestingly, venous valves were not observed in the trapezius, compared to 1-7 valves per one of the other 3 muscles. Abundant venous mergings without valves might form a venous network acting as a reservoir. Therefore, for the trapezius, collateral venous routes such as the external vertebral plexus are essential to provide additional peripheral drainage. However, sufficient muscle-pump function does not seem to be expected for trapezius muscle, given the venous courses parallel to muscle fibers. PMID- 16711164 TI - Histochemical evaluation of angiogenesis in endometrial adenocarcinoma. AB - The formation of new vessels from endothelial cells, termed angiogenesis, is a complex process that is controlled by numerous paracrine factors. Vacularization of tumor tissue is a prerequsite for tumor growth and enables the dissemination of tumor cells throughout the body. Angiogenesis is classically assessed by counting the vessels or clusters of endothelial cells after selective immunohistochemical staining. In this study, vascularization of tumor tissue was evaluated quantitatively in 36 cases of stage I endometrial adenocarcinoma. The goal of this study was to evaluate whether the microvessel density correlates with the invasion potential of endometrial adenocarcinoma. The results show an increase in the number of endothelial cells at different stages in endometrial adenocarcinoma stage I and a strong positive correlation between the endothelial to-stromal ratio and tumor grading. PMID- 16711165 TI - The face-physiognomic expressiveness and human identity. AB - The facial structure develops through the harmonious interaction of bones, mandibular and cervical joints, masticatory, facial and cervical muscles, connective and fatty tissue as well as tracts of conduction. The face represents the personal identity. Ten basic types of faces exist, although they are difficult to classify. No face is symmetrical (biological variance). The expressions are generated by muscles and tension of the elastic facial skin and enable the face to be an organ of expression. The cranial position supports and influences the expression. Certain diseases change the expression in such typical ways that diagnosis and prognosis of some diseases may be concluded from it. PMID- 16711166 TI - Development of the protympanum. AB - The protympanum, the only opening of the tympanic cavity, has enormous significance in the pathogenesis of middle ear inflammation. In our study, we analyzed 70 macerated temporal bones stereomicroscopically to understand the development of the protympanum. Details of this embryonic process were attained using histological sections made in frontal and horizontal planes. In contrast to recent descriptions, the present work reveals that almost the entire protympanum develops from different processes of the petrous bone. The tympanic bone takes part in forming only the posterior border of the lateral wall of the protympanum. Consequently the tympanic plate, which forms the inferior half of the mandibular fossa, is formed by two anlages. The anterosuperior quarter is formed by the petrous bone and the remainder by the tympanic bone. The main structures of the medial wall of the protympanum are the carotid canal with the internal carotid artery (ICA). The carotid canal starts to develop as two laminae of the petrous bone in the 18th fetal week. In cases of incomplete closure of these laminae the carotid canal occupies dehiscences increasing the danger of injury to the ICA. The presence of the ICA is necessary for the development of the carotid canal. If the artery takes an aberrant pathway in the tympanic cavity, or is absent, there is no sign of the carotid canal. PMID- 16711167 TI - Morphometric evaluation of the frontal sinus in relation to age. AB - To investigate the development of the frontal sinus size during life, we studied the planar morphometry in 60 frontal radiographs of patients of different age and gender. A professional software (Bersoft Image 4.02) was used to measure the frontal area of the right and left frontal sinuses on radiographic images. A frontal sinus was already evident in 4-year-old children. Unilateral or bilateral absence of the frontal sinus was seen in 5% of cases. The size of the sinusal area increases up to 19-year-old patients, synchronous with general craniofacial growth. In adults, individual differences in size and shape occurred in relation to environmental factors. In some elderly patients, osseous resorption led to an enlargement of the frontal sinus that might complicate surgical procedures performed in this area. PMID- 16711168 TI - Lost and found anatomical terms: Crista choanalis vomeris. AB - Pars cuneiformis vomeris and Crista choanalis are morphological structures on the vomer listed in the current official anatomical nomenclature, Terminologia anatomica, but are either not mentioned or incorrectly described in different anatomical books. The term Crista choanalis vomeris was originally proposed by Drago Perovic in 1958 to describe the vertical crest beginning at the angle of the alae vomeris, running downwards to the posterior free margin of the vomer. Perovic also described the part of the vomer behind this crest was shaped as a cone, and termed it Pars cuneiformis vomeris. Because of their important contributions to the function of the respiratory pathway, Crista choanalis vomeris and Pars cuneiformis vomeris deserve proper definitions in the anatomical terminology. PMID- 16711169 TI - True cost of guideline implementation. PMID- 16711170 TI - Biophysical effects of repetitive removal of adhesive dressings on peri-ulcer skin. AB - OBJECTIVE: To study the effect of repeated removal of four different adhesive dressings on peri-ulcer skin using quantitative non-invasive techniques. METHOD: Forty-five patients with open (n = 29) or healed (n = 16) venous leg ulcers were included. Peri-ulcer skin was treated for 14 days with patches of two different hydrocolloid-based adhesive dressings, one polyurethane adhesive and one soft silicone adhesive dressing. Normal skin of the patients' ventral forearm was also treated identically. Adhesive patches of the dressings were replaced every second day. The skin barrier function was assessed by measuring transepidermal water loss and stratum corneum hydration by measuring electrical conductance. RESULTS: Thirty-nine patients completed the study. The hydrocolloid adhesives increased transepidermal water loss and conductance while the polyurethane and soft silicone adhesives did not influence these parameters significantly compared with adjacent non-treated peri-ulcer skin. For normal forearm skin, similar relative effects among the four adhesives were found. CONCLUSION: Repetitive treatment with hydrocolloid-based adhesive dressings induced major functional alterations of the stratum corneum. In contrast, a polyurethane adhesive and a soft silicone adhesive dressing did not alter transepidermal water loss or conductance of peri ulcer skin. PMID- 16711171 TI - Evaluation of a self-adherent soft silicone dressing for the treatment of hypertrophic postoperative scars. AB - OBJECTIVE: The primary objective was to compare the efficacy of a self-adherent soft silicone dressing (Mepiform) with 'left-alone' management of hypertrophic scars using theVancouver Scar Scale. Secondary objectives were to follow photographs of the scars, patients' opinions of the scars, and doctors' and patients' assessments of the overall dressing performance, safety and tolerance. METHOD: An exploratory open randomised controlled clinical investigation was undertaken on 11 female patients aged 21-43 years with postoperative scars (nine following breast surgery, two following lower abdominal-glutealplasty). Treatment was initiated between two weeks and two months (mean 4.7 weeks) after surgery. Ten patients completed the 12-month investigation; one patient in the treatment group discontinued for personal reasons. RESULTS: All parameters in the Vancouver Scar Scale improved in both groups, although patients treated with the soft silicone dressing showed greater and more rapid improvements compared with the non-treated patients, while their assessments of the condition of the scar were more favourable. Medical staff rated the overall dressing performance as 'very good' or'good'. One adverse event was reported--local skin irritation at the site of the scar. CONCLUSION: The results suggest that patients treated with the soft silicone dressing experienced greater and more rapid improvements compared with non-treated patients. These results concur with those of previous studies. The fact that Mepiform is self-adhesive and causes limited damage to the stratum corneum on removal gives it an added value compared with non-adhesive silicone gel dressings. PMID- 16711172 TI - Non-healing anal ulcers associated with nicorandil. PMID- 16711173 TI - Effect of a sustained silver-releasing dressing on ulcers with delayed healing: the CONTOP study. AB - OBJECTIVE: To compare the effect of the sustained silver-releasing foam dressing Contreet Foam (ColoplastA/S) with local best practice (LBP) on delayed healing ulcers using a real-life setting. METHOD: A total of 619 patients with ulcers of varying aetiologies were treated for four weeks with either the silver foam dressing or LBP. RESULTS: Wound area was reduced by 50% with the silver foam and 34% with LBP Less slough and maceration, a faster reduction in exudate level and more positive wound progress was achieved with the silver foam. In addition, exudate handling, ease of use, odour and pain improved. Less time was spent on dressing changes, and mean wear time was longer for the silver foam (3.1 days) than for LBP (2.1 days). All differences were statistically significant (p < 0.05). The silver foam dressing outperformed all of the other dressing categories including moist wound healing products and other silver dressings. CONCLUSION: This large-scale comparative real-life study shows that the silver foam dressing supports faster healing of delayed healing wounds. PMID- 16711174 TI - Magnet therapy: opening the debate. PMID- 16711175 TI - Metronidazole chemotherapy. PMID- 16711176 TI - Pulse oximetry and sub-bandage pressure. PMID- 16711177 TI - Can near infrared spectroscopy measure the effect of pressure on oxygenation of sacral soft tissue? AB - OBJECTIVE: To test whether near infrared spectroscopy (NIRS) is applicable for the examination of the influence of external pressure on oxygenation of the soft tissues in the sacral area. METHOD: Tissue oxygenation was measured in 33 healthy volunteers in the prone position. A NIRS probe was positioned over the sacrum and external pressure was applied in 10 mmHg increments, from 20 mmHg to 200 mmHg and then decreased. At each level, tissue oxygen saturation (StO2) was measured. To test reproducibility, the protocol was repeated in six volunteers, in whom the thickness of the soft-tissue envelope at different levels of external pressure was assessed using ultrasound. RESULTS: There was wide variability in StO2 courses between the 33 subjects, with a non-linear relationship between pressure and StO2. The only consistent finding was that the StO2 was significantly higher after decreasing pressure than at the initial pressure of 20 mmHg, which is indicative of reactive hyperaemia. Despite the application of high external pressures, reasonable tissue oxygenation was maintained in 19 of 33 subjects. Reproducibility of the measurements was poor. Comparison of soft-tissue thickness with corresponding StO2 values showed that, with increasing pressure, the percentage decrease in tissue thickness was higher than the decrease in tissue oxygenation. CONCLUSION: This study confirms that NIRS is not useful for assessing tissue oxygenation in pressure ulcer research due to unacceptable inter individual variability and poor reproducibility of measurements. PMID- 16711178 TI - Use of a bioengineered human dermis on a large acute foot ulcer. PMID- 16711180 TI - Code of ethics for nurses at a European level. PMID- 16711179 TI - Effect of enzymatic debridement with collagenase on acute and chronic hard-to heal wounds. AB - OBJECTIVE: A large retrospective analysis was performed to assess the outcomes of burns and chronic ulcers treated with collagenase in an outpatient setting. No cost comparison was performed. METHOD: Of 979 patients entered into the analysis, 647 had burns affecting < or = 15% body surface area (BSA) and 332 had chronic ulcers of various aetiologies. AII were treated with collagenase-based ointments once daily (Noruxol or Iruxol, Smith and Nephew). Treatment was continued until complete healing was achieved. RESULTS: In burns patients the overall average healing time was 17.9 days in the paediatric population and 23.6 days in adults. Burn depth and presence of eschar were the main factors affecting healing probability. The chronic ulcers were predominantly of diabetic, venous and mixed aetiology. Average healing time was 15.4 weeks, with ulcers of mixed aetiology showing the shortest average healing time (9.2 weeks). There was a positive correlation between wound area and healing time. The topical application of collagenase-based ointments was well tolerated by patients and caregivers. CONCLUSION: This large retrospective analysis shows that collagenase treatments in outpatient clinics are effective and well accepted in patients with burns affecting < or = 15% BSA or with chronic ulcers of various aetiologies. Implementation of collagenase treatments in outpatient clinics has the potential to improve wound healing and may also decrease the cost of wound care. PMID- 16711181 TI - Is H5N1 like a ghost flu that can cause a pandemic in humans? PMID- 16711182 TI - Cancer patients' perception of being or not being confirmed. AB - The aim of this study was to obtain in-depth knowledge about caring confirmation of patients with cancer, from the patients' point of view. The research topic was: what is the significance for patients of their being confirmed by nursing personnel? Fifteen men and women between 43 and 80 years of age participated in this study. The method of data collection used was qualitative research interviewing. A hermeneutic approach was used to interpret the data, in which Kvale's self-perception, the 'common sense' level, and theoretical levels were applied. The results are summarized in three areas: an outer confirmation, an inner confirmation, and a lack of the latter. Outer confirmation meant being understood and taken seriously; the maintenance of human dignity and worth indicated inner confirmation. A lack of inner confirmation is primarily manifest in terms of patients' mental, spiritual and existential concerns. In relation to the theory of Eriksson, these patients were confirmed at the level of having and being, but seldom at the level of becoming. PMID- 16711183 TI - Patients' dignity in a rehabilitation ward: ethical challenges for nursing staff. AB - The purpose of this study was to explore the challenges met by nursing staff in a rehabilitation ward. The overall design was qualitative: data were derived from focus interviews with groups of nurses and analyzed from a phenomenological hermeneutic perspective. The main finding was that challenges emerge on two levels of ethics and rationality: an economic/administrative level and a level of care. An increase in work-load and the changing potential for patient rehabilitation influence the care that nurses can provide in rehabilitating patients, and therefore also affect patients' feelings of self-worth and dignity. Some patients wish to maintain their independence and autonomy, whereas others seem to 'lose themselves'. Independence and autonomy are associated with dignity, but their lack is contrary to it. PMID- 16711184 TI - Everyday ethics in the care of elderly people. AB - This article analyses the general ethical milieu in a nursing home for elderly residents and provides a decision-making model for analysing the ethical situations that arise. It considers what it means for the residents to live together and for the staff to be in ethically problematic situations when caring for residents. An interpretative phenomenological approach and Sandman's ethical model proved useful for this purpose. Systematic observations were carried out and interpretation of the general ethical milieu was summarized as 'being in the same world without meeting'. Two themes and four subthemes emerged from the analysis. Three different ethical problems were analysed. The outcome of using the decision-making model highlighted the discrepancy between the solutions used and well-founded solutions to these problems. An important conclusion that emerged from this study was the need for a structured tool for reflection. PMID- 16711186 TI - Bridging Western ethics and Japanese local ethics by listening to nurses' concerns. AB - Among Japanese nurses ethics is perceived as being distant and unrelated to their practice, although this is filled with ethical concerns and the making of ethical decisions. The reasons for this dissociation are the primacy of western values in modem Japanese health care systems and the suppression of Japanese nurses' indigenous ethical values because of domination by western ethics. A hermeneutic study was conducted to listen to the ethical voices of Japanese nurses. Seven ethical concerns were revealed. Although some of these concerns may seem to share similar values with western ethical principles, the basis for the concerns was unique and rooted in the Japanese cultural value system. The meanings of each concern are explicated in conjunction with related background meanings. Listening and trying to understand these nurses' voices in their own context suggests a way of bridging the gap between abstract and universal ethics and practical and local ethics. PMID- 16711185 TI - Nurses' attitudes towards people with dementia: the semantic differential technique. AB - One important aspect of the nurse-patient relationship is nurses' attitudes towards their patients. Nurses' attitudes towards people with dementia have been studied from a wide range of approaches, but few authors have focused on the structure of these attitudes. This study aimed to identify a structure in licensed practical nurses' attitudes towards people with dementia. Twenty-one group dwelling units for people with dementia at 11 nursing homes participated in the study. A total of 1577 assessments of 178 patients were sent out to 181 respondents and 1237 answers were returned. The semantic differential technique was used. The scale had 57 bipolar pairs of adjectives that estimate an unknown number of dimensions of nurses' attitudes towards an identified patient. The assessments were analysed using entropy-based measures of association combined with structural plots. The analysis revealed four dimensions, which related to licensed practical nurses' opinions of the patients: an ethical and aesthetic dimension; an ability to understand; an ability to experience; and an ability for social interaction. The results of the study indicated that, on the positive to negative attitude continuum, the nurses' attitudes fell at the positive to neutral end. This is an important finding owing to the personhood perspective, from which it is reasonable to assume that, with a more positive attitude to people with dementia, the prerequisites for person-centred care will improve. PMID- 16711187 TI - Decision making in acute care: a practical framework supporting the 'best interests' principle. AB - The best interests principle is commonly utilized in acute care settings to assist with decision making about life-saving and life-sustaining treatment. This ethical principle demands that the decision maker refers to some conception of quality of life that is relevant to the individual patient. The aim of this article is to describe the factors that are required to be incorporated into an account of quality of life that will provide a morally justifiable basis for making a judgement about the future quality of life, and therefore the best interests, of critically ill patients who are mentally incompetent. This account consists of three major components pain and suffering, body functioning, and autonomy and is applicable in situations where very limited information is available to guide decision making. This framework helps to make decisions about the provision of life-saving treatment that are as consistent as possible in all patient situations. PMID- 16711188 TI - Stem cell and related therapies: nurses and midwives representing all parties. AB - Nurses and midwives are part of health care in all the stages of our lives from preconception to death. Recent scientific advances have introduced new techniques of screening and diagnosis linked to stem cell isolation and therapies. These could affect us at any age and therefore nurses will be involved as carers and patients advocates for these techniques. In this article stem cell techniques and therapies are outlined, as well as some of the ethical challenges faced by various nursing groups, whether in adult, learning difficulties, mental health, paediatric, primary care, public health or health visiting areas. Nurses have to understand the ethical issues and the rights of all parties (donor, recipient and families), which may conflict with each other, to be able to weigh up the benefits and costs to each group involved. Answers have to be found on a case-by case basis within local moral frameworks and law. Nurses represent all parties in these therapies and act as advocates for every patient group. They need to act in an interprofessional environment to promote the best interests of all their clients, both clinically and ethically. PMID- 16711189 TI - Cultural and historical perspectives on nursing and ethics: listening to each other--report of the conference in Taipei, Taiwan, 19 May 2005, organized by ICNE and Nursing Ethics. PMID- 16711190 TI - Passive euthanasia. PMID- 16711191 TI - [Itching, discharge, aches, bleeding. Problems in the bottom area]. PMID- 16711192 TI - [Chronic recurrent diverticulitis. Surgeons in retreat]. PMID- 16711193 TI - [Metabolic syndrome threatens to blow up the health care system. Warning before the great detonation]. PMID- 16711194 TI - [Colic-like abdominal pain and edema in a young woman. Cause of complaints was not in the abdomen]. PMID- 16711195 TI - [Early-summer meningoencephalitis cases increase in Europe. Vaccination time is now!]. PMID- 16711196 TI - [Tuning fork study in deafness. Thus function Rinne and Weber tests]. PMID- 16711197 TI - [Severe incidents in phase I clinical trial. How dangerous is the step from animal to man?]. PMID- 16711198 TI - [Even in 60-year-old atrial fibrillation is not uncommon. Advances in long-term treatment]. PMID- 16711199 TI - [Drug treatment of atrial fibrillation]. AB - Before initiating specific treatment of atrial fibrillation treatable causes must first be reliably ruled out. Furthermore, the chances of maintaining a sinus rhythm must be individually weighed against the potential complications and risks of an anti-arrhythmic therapy, and also alternative strategies considered. Today, rate control is recommended in the case of asymptomatic patients, in particular in the elderly, while rhythm control is the strategy of choice in younger, symptomatic patients. For recurrence prevention, beta blockers are the first choice drugs. In patients with no structural heart disease, class IC antiarrhythmic agents, in those with structural heart disease only amiodarone, may be considered. Promising new additive therapeutic approaches are ACE inhibitors and AT-II receptor antagonists. In the future, combinations of pharmacotherapy and non-drug treatments will help to improve the treatment of atrial fibrillation. PMID- 16711200 TI - [Prevention of thromboembolism in patients with atrial fibrillation]. AB - Provided that account is taken of the criteria discussed in the present article, there is no doubt about the therapeutic benefits of effective anticoagulation in patients with chronic atrial fibrillation. Indeed, it is to be expected that the previously valid therapeutic guidelines are more likely to be expanded to reduce feared thromboembolic complications to a minimum, as is exemplified by the recommendation that the application of anticoagulation treatment with vitamin K antagonists should be continued over the longer term, that is, after the restoration of sinus rhythm. Furthermore, there is hope that effective drugs with a calculable (level of) safety and simplicity of administration may soon become available. PMID- 16711201 TI - [Catheter ablation of atrial fibrillation]. AB - In patients with drug-refractory atrial fibrillation, left-atrial catheter ablation represents a new curative therapeutic option. Segmental ostial or circumferential pulmonary vein isolation can achieve stable sinus rhythm in some 70% of patients with paroxysmal atrial fibrillation but no severe structural heart disease. In patients with chronic atrial fibrillation, complex left-atrial linear, or substrate-oriented ablation strategies may additionally be applied. In patients with cardiac insufficiency or more severe systolic left-ventricular dysfunction, restoration of a stable sinus rhythm through the use of left-atrial catheter ablation can improve the left-ventricular ejection fraction and reduce the severity of cardiac failure. Potential complications of ablation include, in particular, pulmonary veins stenosis, iatrogenic left-atrial tachycardia, thromboembolic events and fatal atrio-esophageal fistulas. PMID- 16711202 TI - [A 15-year-old boy with spontaneous pain in the lower leg]. AB - On kicking the ball during a football tournament, and with no involvement of any other player, a 15-year-old boy experienced a spontaneous fracture of the tibia with no fragment displacement. The X-ray of the region of the fracture revealed a cystic structure measuring approximately 3 cm x 2 cm x 2 cm. After eight weeks of conservative treatment, radiography showed bony consolidation of the cyst, and the patient was pain-free. The differential diagnosis must give consideration to juvenile and aneurysmal bone cysts, giant cell tumors and malignant tumors (e.g. osteosarcoma and Ewing's sarcoma). PMID- 16711203 TI - [Irrespective of their educational status, patients with insulin-dependent type 2 diabetes mellitus benefit from patient-centered medical behavioral training]. PMID- 16711204 TI - [Stretching financial possibilities of practice. Cooperation: as good as 15% more patients]. PMID- 16711205 TI - [To detect iodine deficiency goiter early. What does palpation bring?]. PMID- 16711206 TI - [Every third employed person affected. Screening is federally worthwhile]. PMID- 16711207 TI - [Lower leg edema. Persistent fluid accumulations have always to be clarified!]. PMID- 16711208 TI - [Medicine in literature: who has written it? Comforting preparations for planned suicide]. PMID- 16711209 TI - [Cave: child's large eye]. PMID- 16711210 TI - Toward measuring visualization insight. PMID- 16711211 TI - Teaching communication skills with virtual humans. PMID- 16711212 TI - Computer graphics advances the art of anime. PMID- 16711213 TI - Interactive narrative. PMID- 16711214 TI - From linear story generation to branching story graphs. PMID- 16711215 TI - Declarative optimization-based drama management in interactive fiction. PMID- 16711216 TI - Unscripted narrative for affectively driven characters. PMID- 16711217 TI - Generating comics from 3D interactive computer graphics. PMID- 16711218 TI - Multimodal interaction with a wearable augmented reality system. PMID- 16711219 TI - Cords: geometric curve primitives for modeling contact. PMID- 16711220 TI - Isn't it about time? PMID- 16711221 TI - How to solve a cubic equation, part 1: the shape of the discriminant. PMID- 16711222 TI - Kodak may exit health IT. Changes to healthcare group may make buyers leery. PMID- 16711224 TI - Outlook remains bleak. Earlier bankruptcy seen for Medicare trust funds. PMID- 16711223 TI - Transparency with a catch. AHA urges hospital price disclosure, but wants protection from lawsuits and broad definition of community benefits. AB - The AHA last week added its two cents to the transparency debate, approving a policy that supports a patient's right to know pricing. James Springfield, left, CEO of Valley Baptist Health System, who backs the AHA's policy, says the changes come at a time when "the public is clamoring ... to learn more about the mystical world of healthcare. PMID- 16711225 TI - Rule change: payday or mayday? Uncertainty looms as hospitals scramble to get data. PMID- 16711226 TI - Top healthcare REIT grows, Health Care Property's value to hit dollar 11.5 billion. PMID- 16711227 TI - Tenet spotlights outpatient care. Plans to meet demand, competition under way. PMID- 16711228 TI - Satisfied sans good service. Hospital reviews good, despite low-quality care. PMID- 16711229 TI - In case of emergency. Hospitals weigh pandemic supply options. PMID- 16711230 TI - La. asks for safety network. PMID- 16711231 TI - Illinois joins GPO game. Purchasing unity is goal of new state collaborative. PMID- 16711232 TI - Seeking allies. Not-for-profits partner with groups. PMID- 16711234 TI - The Baron of Healthcare. With 46 million uninsured, it's time for billionaire McGuire to open his coffers. PMID- 16711233 TI - AAFP denies endorsement. Deal with MinuteClinic will be 'advisory' only. PMID- 16711235 TI - No news is not good news for attendees. World Health Care Congress offered everything but breaking headlines. PMID- 16711236 TI - Devising new tactics. Providers and medical device manufacturers struggle to find best ways to work together on recalls, advisories. PMID- 16711238 TI - Disciplinary actions dropped in 2005. Group concerned that numbers don't accurately reflect physicians' records. PMID- 16711237 TI - Hey, where's the junk food? Drug vending machines offer convenience, but safety, security issues raised. PMID- 16711239 TI - [Computed tomography in the evaluation of cochlear structural features in cochlear implantation]. AB - Temporal bone computed tomographic scans were studied in 136 deaf patients of different age and sex, including 56 patients (39 children and 17 adults) rehabilitated with cochlear implantation (CI). The mean values of the cochlear parameters that are important for the results of a surgical CI stage, namely: the diameter of the cochlear base (DCB), the diameter of the basal helix, the height of the cochlea, the angle between the cochlear base and the promontorial wall, were calculated. These parameters were compared in different age groups. A correlation was determined for each of the studied parameters and for the depth of intracochlear insertion of electrodes. There was a significant correlation between DCB and the depth of insertion of the electrodes of CI22M and CI24M cochlear implants. DBC at which the active part of the electrode chain of an implant could be significantly more frequently placed in the tympanic scala of the cochlea was estimated. PMID- 16711240 TI - [Role of spiral computed tomography in the diagnosis of complications due to endodontic treatment]. AB - The paper deals with the diagnosis of some errors in endodontic treatment by spiral computed tomography. PMID- 16711241 TI - [Ultrasound and morphological diagnosis of thyroid diseases in radiation-exposed persons]. AB - The purpose of the present investigation was to enhance the efficiency of diagnosis of thyroid diseases in irradiated persons. Three hundred persons exposed to radiation in doses of 5 to 250 SeV and the postexposure period was 9 to 48 years. A control group comprised 100 military men unexposed to ionizing radiation. Thyroid ultrasound study and morphological diagnosis revealed diffuse nodular goiter transformation in 66% of the irradiated persons and malignant thyroid neoplasms in 3.7% whereas 38% of the control patients were found to have only benign diffuse nodular thyroid alterations and autoimmune thyroiditis (AIT). The investigation identified the echosemiotic features of nodular masses and AIT in the study group patients and established their association with the pathomorphological changes of the thyroid. The authors note the high efficiency of complex clinicoradiation and morphological studies in detecting thyroid pathology and the priority in establishing an accurate diagnosis at the preoperative stage and in determining management tactics. PMID- 16711242 TI - [Pulmonary venous occlusive disease]. AB - The present study deals with the radiation diagnosis of the rare disease- pulmonary venous occlusive disease. The follow-up covered three cases that ended with death. The clinical picture of the disease did not differ from the manifestations of primary pulmonary hypertension. All the patients underwent chest X-ray study in four standard projections. The morphological verification of its diagnosis was made on the basis of autopsy data. X-rat study promoted identification of such signs as peculiar changes in the lung pattern in form of its looping, reticulation, fine-focality along with reticular changes, the presence of Kerley lines, the diameter of root branches, enlargements of the pulmonary trunk without any symptoms of the enlarged left atrium. PMID- 16711243 TI - [Potential use of current X-ray contrast studies in the detection of small intestinal polyps in the Peutz-Jeghers syndrome]. AB - The paper presents the results of studies of the small bowel in 9 patients with the Peutz-Jeghers syndrome. The indications for the study were the clinical picture of gastrointestinal bleeding and the symptoms of ileus. The author's intubation enterographic procedure using the better composition of barium suspension and an infusion system for administering contract substances into the small bowel, an improved fractional contrasting procedure, and a procedure employing the agent Entero-view was applied for contrasting the small bowel. All the procedures revealed an obvious picture of polyposis of the small bowel. The minimum size of detected tumors was 0.3-0.4 cm in diameter. Emphasis is laid on the quantitative image of neoplasms with Entero-view and the therapeutic effect of intubation enterography in evolving small-small intestinal intussusception. PMID- 16711244 TI - [Potential use of radiation diagnosis of esophageal achalasia]. AB - The prime objective of the paper is to attempt to reevaluate the potentialities of current radiation diagnosis of esophageal achalasia. Moreover, its main idea is to assess the procedure of a traditional X-ray section of radiation study. In the authors' opinion, the main motivation of writing this paper was the emergence of new recent potentialities of radiation diagnosis, including its traditional X ray section in gastroenterological pathology, as well as a considerable body of information on esophageal achalasia (144 cases). The tasks to be solved in the paper include the differential diagnosis of esophageal achalasia and cardioesophageal cancer, as well as the basic capacities of radiation diagnosis to help clinicians in characterizing the changes caused by medical and surgical treatments for this disease. The authors consider that the obtained volume of necessary information on this abnormality rather frequently encountered in esophagogastroenterological pathology can be increased by applying the current possibilities of radiation study and mainly its traditional X-ray section. PMID- 16711245 TI - [Densitometric parameters in the presence of risk factors and complications of osteoporosis]. AB - The paper deals with the densitometric study of 3000 persons due to 21 to 85 years. It presents the major common risk factors of osteoporosis and osteopenia and the indices of bone mineral density in relation to age, risk factors, and a history of bone fractures. Groups of the examinees were formed according to the history and densitometric data. Based on the findings, the author has substantiated a methodological approach to using X-ray densitometry in the identified groups. PMID- 16711246 TI - [Current methods of radiation diagnosis and surgical intervention in cervical and upper thoracic spine and spinal cord]. PMID- 16711247 TI - [Clinical and radiological studies in catamnesis of the patients, treated for ovarian cancer (review of literature)]. PMID- 16711248 TI - [The 11-th European Symposium on Urogenital Radiology]. PMID- 16711249 TI - [Modes for enhancing the efficiency of gadolinium chelates as contrast substances]. PMID- 16711250 TI - Screening of carbohydrate-specific phage antibodies against recombinant human erythropoietin (rhuEPO) using a phage display antibody library: preliminary study. AB - This paper is a preliminary report on development of a screening method for carbohydrate-specific phage antibodies against recombinant human erythropoietin (rHuEPO), using a phage display antibody library. rHuEPO was oxidized with sodium periodate or treated with 1,4-dithiothreitol and guanidine hydrochloride for detecting the specificity of obtained phage antibodies. Of 100 phage clones, three initially showed higher carbohydrate-related specificity. One of them (No. 62) bound specifically to the carbohydrate chains of rHuEPO, while the other two (Nos. 63 and 83) might recognize the steric conformation related to both the carbohydrate and the polypeptide chain of rHuEPO. These phage antibodies may serve as useful capture ligands for future development of efficient analytical methods for rHuEPO. PMID- 16711251 TI - Proposed reference material for human free immunoglobulin light chain measurement. AB - Human free immunoglobulin light chain (FLC) kappa and lambda are useful clinical markers for light chain myeloma and AL amyloidosis. With the recent development of specific and reliable FLC immunoassays, the quantitative measurement of FLCs will be widely used in clinical practice. However, researchers have used various calibrators, mainly monoclonal FLCs; thus, no standardization has been performed among the assay methods. This prompted us to purify intact FLCs from the pooled urine specimens of healthy volunteers as the first reference materials for FLC assays. After precipitation with ammonium sulfate, FLCs were purified by the following steps of chromatography; cation exchange, gel filtration, and antibody assisted immunoaffinity. SDS-PAGE and Western blotting analyses showed that the purity of FLC kappa and FLC lambda was more than 98%. These purified FLCs did not contain the other immunological types of light chains. These intact and purified FLCs are suitable as the first reference materials to standardize FLC assays. PMID- 16711252 TI - A new competitive enzyme linked immunosorbent assay (MRP83-CA15-3) for MUC1 measurement in breast cancer. AB - A new competitive enzyme linked immunosorbent assay was developed in this study. Monoclonal antibody (PR81) against the tandem repeat of the core protein was prepared, characterized, purified, and conjugated to HRP. This antibody exhibited no cross reactions with proteins such as bovine serum albumin, keyhole limpet homocyanin, human serum albumin, casein, human milk fat globin (HMFG), and peptone. The native cancerous MUC1 protein was purified from ascites fluid of a patient suffering from small cell lung carcinoma by immunoaffinity chromatography and used as a standard preparation in the assay buffer. The standard curve was constructed following a competitive procedure in the range of 0-200 U/mL. The level of MUC1 in normal and cancerous samples was compared following this procedure and using available CA15-3 EIA (Can Ag), as well as LIAISON CA15-3 commercial kits. The correlation coefficient between the procedure reported in this work (MRP83-CA15-3) and CA15-3 EIA (Can Ag) was 0.68 and was 0.95 with the LIAISON CA15-3 kit. We concluded that the present assay can detect MUC1 in breast cancer patients with great sensitivity and accuracy. PMID- 16711253 TI - Studies on the influence of the structural modifications in the tracer on the immunoassay of progesterone. AB - The main objective of the present study was to examine the influence of different bridges in radioiodinated tracers on the assay performance of progesterone using antibodies. Three homologous and two heterologous immunoassay systems for the measurement of progesterone in human serum are described. Using an antiserum raised against progesterone-11alpha-hemisuccinate-bovine serum albumin (BSA), assays with homologous radioligands, namely progesterone-11alpha-hemisuccinate 125I-tyrosine methyl ester (TME) and progesterone-11alpha-hemisuccinate-125I histamine, heterologous bridge radioligand, namely progesterone-11alpha hemiphthalate-125I-TME, and a heterologous site radioligand namely progesterone-3 (O-carboxymethyl) oxime (CMO)-125I-histamine were optimized. A homologous assay system, using antiserum raised against progesterone-3-carboxymethyl oxime-BSA and progesterone-3-CMO-125I-histamine as the radioligand was also optimized to develop a radio-immunoassay (RIA) for serum progesterone. Amongst the two homologous radioligands, viz., progesterone-11alpha-hemisuccinate-125I-histamine and the corresponding TME conjugate tracer, the former yielded a standard curve with a higher slope (-0.6) as compared to the latter (-0.5). The heterologous bridge system with progesterone-11alpha-hemiphthalate-125I-TME resulted in a more sensitive assay (slope of -0.8) than the homologous tracers, whilst the heterologous site radioligand, viz., progesterone-3-CMO-125I-histamine gave the most sensitive assay (slope of -1.2). The homologous assay with antiserum against progesterone-3-CMO-BSA and progesterone-3-CMO-125I-histamine tracer gave a standard curve having a slope of -0.97. The two antibodies developed against progesterone, viz., progesterone-11alpha-hemisuccinate-BSA and progesterone-3-CMO BSA were characterized for their titre, sensitivity, and specificity. Considering the slope, sensitivity, cross-reactivity, and the quality of tracer, the assay system using antiserum against progesterone-11alpha-hemisuccinate-BSA and progesterone-3-CMO-125I-histamine was found to be suitable for the development of RIA for serum progesterone. The bridges used in an immunogen for production of antibodies, as well as in the preparation of tracer, have a great influence on the assay characteristics. PMID- 16711254 TI - Development of a normal human immunoglobulin G standard curve for enzyme-linked immunosorbent assay: use for comparison of antigen efficacy. AB - Internal standard reference curves are used in enzyme-linked immunosorbent assay (ELISA) plates to control for inter- and intra-assay variance. To compare the diagnostic potential of multiple T. solium antigens on an unbiased, universal scale, we have created a standard curve using normal, human immunoglobulin G (hIgG). The hIgG curve is inexpensive and simple to prepare, and remains stable at 5 degrees C for at least one year, with a coefficient of variance of less than 10%. The hIgG standard curve has proven a critical tool for the comparison of several diagnostic antigens slated for assay development. PMID- 16711255 TI - Optimization, comparison, and application of colorimetric vs. chemiluminescence based indirect sandwich ELISA for measurement of human IL-23. AB - Currently, there is neither a published ELISA method nor it is clear whether chemiluminescence substrates would provide better sensitivity vs. colorimetric substrates for measuring human IL-23-a recently described Type-1 immunity associated cytokine. Initially, we optimized a colorimetric ELISA using p-nitro phenyl phosphate substrate. Subsequently, we compared it with chemiluminescence substrates that provided approximately 5-fold enhanced sensitivity (mean sensitivity; 26.3 pg/mL vs. colorimetric assay, 131 pg/mL; p < 0.01). Both methods were reliable, with <10% inter- and intra-assay variations. We then found that the chemiluminescence method was useful in situations where human IL-23 was not readily measurable by a colorimetric method. PMID- 16711256 TI - Correlation between pharmacological efficacy of cyclosporine A and tacrolimus, evaluated by lymphocyte immunosuppressant-sensitivity test (LIST) with MTT assay procedure in renal transplant recipients. AB - The dose of calcineurin inhibitors in renal transplantation has been adjusted, based on the therapeutic drug monitoring data. However, the data do not always correlate with clinical drug efficacy. In vitro response of peripheral-blood mononuclear cells to immunosuppressive drugs is reported to correlate with the recipient-response to therapeutic efficacy of the drug. We report, here, usefulness of a lymphocyte immunosuppressant sensitivity test for the estimation of individual drug sensitivity in renal transplant recipients. The LIST we have developed includes MTT assay procedures without the use of radioisotope-labeled compounds, which is convenient for general hospital use. Utilizing this procedure, we compared the pharmacological efficacy between cyclosporine A and tacrolimus in 41 renal transplant recipients. PMID- 16711257 TI - [Peripheral arterial disease]. AB - The peripheral arterial disease (PAD) accounts for very significant medical issue in the world. Various clinical disorders due to coronary, cerebrovascular, abdominal and obliterative arteriosclerosis of extremities can be classified as the most important medical challenges. This review summarises the etiopathogenesis of this systemic problem including the function of endothelium as well as the anatomic, physiologic relevance, and major risk factors of atherosclerosis (diabetes mellitus, hypertension, obesity, smoking etc.). The first step is the dysfunction of endothelial monolayer, later on the plaques caused by humoral and cellular reactions can produce abnormal blood flow. The atherosclerosis in relation to plaque rupture with intraarterial thrombosis constitutes acute and chronic circulation disturbances and tissue damage. Diagnostic procedures are also presented regarding not only the symptomatic, but focusing on the early, asymptomatic phase of the disease, as well. By giving full details of the acute and chronic treatment the author emphasizes its basic concepts. He also points out the importance of evidence based medicine. PMID- 16711258 TI - [The role of free radicals in the UV-induced skin damage. Photo-aging]. AB - The natural (intrinsic) ageing of the skin is enhanced by environmental factors (extrinsic ageing). One of the most important exogenous factors is the solar UV exposure, which results in photo-aging. Besides this, epidemiological and experimental data show a rapid increase in the incidence of human skin cancers, which is also in relation to the increased sunlight exposure of the skin. In the background of these processes there are cell biological effects, photochemical reactions, membrane receptor changes, lipid- and protein modifications, DNA damage induced by UV. The qualities and quantities of them are wavelength dependent. The UVB photons are absorbed mostly by the DNA of the epidermal keratinocytes, therefore this spectrum is more relevant for photocarcinogenesis. The effect of UVA-irradiation is mainly manifested in the induction of free radicals, which have not only DNA-damaging, but also immunomodulating effect, which also can influence on tumour development. Furthermore, the free radicals cause dermal connective tissue damage as well via activating transcription factors, inducing matrix metalloproteinases, diminishing the procollagen I and fibrillin-1 synthesis. These processes are augmented by mitochondrial DNA mutations, protein oxidation, apoptosis induction. Therefore the enzymes neutralising free radicals and antioxidant molecules, respectively, have an important role in the defence mechanisms. In the therapy of photo-aging the local retinoids lived up to expectations, but the clinical effectiveness of antioxidant vitamins is lower than expected. The most important factor in the prevention of the photo-aging and photocarcinogenesis is the sun protection at present. PMID- 16711259 TI - [Effects of environmental noises and sounds on the quality of life controlled by the central nervous system]. AB - From the environment surrounding us, there are different sounds, noises as physical waves, which are arriving to the living organism. These dissonant and consonant inputs have a negative or a positive effect on the quality of life, respectively. The noise, as a dissonant sound has always a negative effect, while music, in the majority of cases, a positive effect. Both the professional and the non-professional noise as a pollution by its stress-provoking effect seriously damages the health. Therefore there is an important endeavour fighting against noise pollution to improve the quality of life. Well-selected music, by means of neurochemical transmissions, strengthens the immune system of the brain in all periods, from the intrauterine period to the end of life--it has a positive effect on the quality of life. PMID- 16711260 TI - [Change in location of colorectal cancer in Hungarian patients between 1993 2004]. AB - The incidence of proximal tumours in Western countries has steadily increased while that of distal tumours has shown a corresponding decrease. Our aim was to investigate the prevalence, location and histology of colorectal cancers in the last twelve years in Hungarian patients. PATIENTS AND METHODS: Clinical data of 1738 patients diagnosed with colorectal tumors (M/F: 940/798, mean age at diagnosis: 65.2 +/- 12.5 years) between 1st of January 1993 and 31st of December 2004 at the 1st Internal Medicine and 1st Surgery Department of Semmelweis University were enrolled. Pathology and clinical data were analysed retrospectively. The observed periods were the following 1993-1998 and 1999-2004. RESULTS: 1694 (97.5%) of the patients had adenocarcinoma (CRC), 15 anaplastic cancers, 9 carcinoid, 6 planocellular, 5 GIST, 3 leiomyoma and 2-2 melanoma, lymphoma and shigillocellular cancers were diagnosed. 75.7% (943/1246) of the CRCs were diagnosed at locally advanced stage (T3-T4), and 47.7% (521/1093) of CRC patients had lymph node metastasis at the time of diagnosis. 11.0% of the CRCs were diagnosed in <50 year-olds (<40 years: 2.5%, <30 years: 0.5%). The location of the CRC was distal in 1186 (rectum: 53.9%, sigmoid/descending: 46.1) and proximal in 508 cases. Synchronous cancers were detected in 12 patients (age: 68.8 +/- 11.6 years, gender: 11 male/1 female, location: rectum and transverse in 6, rectum and ascending/caecum in 5 patients). Age at diagnosis was not different according to gender (M/F: 64.8 +/- 12.0 years vs. 65.8 +/- 12.9 years), but it was lower in patients with rectal cancer compared to left or right sided cancers (64.1 years vs. left: 66.1 years, right: 66.0 years, p = 0.02). Rectal CRC was more common in males, while the proportion of proximal cancers was lower (rectum, M/F: 41.2% vs. 33.5%, proximal M/F: 26.8% vs. 33.8%, p = 0.003). The proportion of rectal cancers increased over the observed period (1993-1998: rectal: 31.6% vs. 1999-2004: 42.1%, p = 0.002). CONCLUSIONS: In contrast to Western countries, the proportion of proximal CRC did not become higher in Hungary. Still more than two-third of the patients were diagnosed to have distal cancers. The proportion of male patients was higher in this subset of CRC. The high percentage of locally advanced and metastatic cancers supports the need for colorectal screening program in Hungary. PMID- 16711263 TI - [Remembering Arpad Csapo (1918-1981)]. PMID- 16711261 TI - [Recent changes in concepts of antioxidant treatment]. AB - The promising theoretical possibilities of antioxidant prevention and protection against vascular diseases and neoplasms could not have been realized as yet. The author searches into the causes of this failure by analyzing data of recent literature. Previous preventive trials as well as newly discovered pharmacological and molecular biological effects of antioxidants are reviewed. Results of meta-analyses on prevention trials of vascular disease by vitamin-E and those of gastrointestinal cancers are also included. The lately recognized properties of antioxidants are surveyed with special regard to their capability of modulating apoptosis, inducing gene expressions and their transformation into pro-oxidants. The harmful consequence of high doses of a single antioxidant is emphasized. The retinoids, vitamins D and K possess both pro-apoptotic and antiproliferative activity, while N-acetylcysteine exerts mainly anti-apoptotic effects. Since the effects of the eight vitamin E homologues are different in many respects, alpha-tocopherol can not be regarded as vitamin E of full value. Antioxidant supply from natural sources does not seem to be sufficient for an adequate preventive effect. The author recommends such a combination in which physiological amounts of vitamins C, D, K and B-complex, N-acetylcysteine, vitamin E of natural origin might be complemented by allopurinol, co-enzyme Q-10 and alpha-lipoic acid. A diet rich in flavonoids and carotenoids is essential. Application of appropriate laboratory methods is of great value in the individualization, monitoring and control of antioxidant treatment. PMID- 16711262 TI - [Treatment of colitis ulcerosa and Crohn's disease]. PMID- 16711264 TI - [Aladar Toth M.D., ear-nose-throat specialist, head of the ENT-department (1903 1976)]. PMID- 16711265 TI - [Samuel Kazzay and the first pharmacy in Debrecen]. PMID- 16711267 TI - TMD? TMD? What ever happened to the tops of the teeth? PMID- 16711266 TI - How many years must a profession exist...? PMID- 16711268 TI - Dr. Melis comments on Simmons "Guidelines . . ." which appeared in the October 2005 issue of CRANIO. PMID- 16711269 TI - Asymmetry of the articular eminence in dentate and partially edentulous populations. AB - The purpose of this study was to assess the degree of right-left asymmetry of the glenoid fossa. The specific aims were (1) to determine whether there were relationships between age, number of teeth, slope of the articular eminence, fossa depth, and the degree of right-left asymmetry, and (2) to compare the right left asymmetry of two populations, one characterized by an acceptable occlusion (A-Occ), the other by an unacceptable occlusion (partially edentulous; Un-Occ). A Occ was defined as possessing a minimum of 28 teeth that would allow for hand articulation of the mandibular teeth to the maxillary teeth. Un-Occ had fewer than 17 teeth, which would make it impossible to articulate the mandible with the maxilla. The sample included 20th century dry skulls: 70 African-American (44 male, 26 female) and 64 European-American (49 male, 15 female), ranging in age from 21-105 (mean 47.1 +/- 19.9). The medial (M), central (C), and lateral (Lat) aspects of the right (R) and left (L) slopes of the articular eminence were measured in a sagittal plane. The R and L fossa depth also were measured. The raw absolute differences IR-LI and relative differences [IR-LI/IR+Llx100] of the articular slope angles (M, C, and Lat) and fossa depths were computed. Statistical analysis included paired t-tests, independent t-tests, and Pearson's correlation coefficients, significance at p < or = 0.05. Ninety percent (90%) of the population exhibited right-left asymmetry of the glenoid fossa. The right articular slopes (M, C, and Lat) were significantly steeper than the left articular slopes; the right fossa depths were significantly deeper than the left. There generally were no differences in the articular slope steepness or fossa depths between the partially dentate and the dentate, nor were there statistical differences between the two groups in the raw absolute differences or relative differences of the M, C, and Lat slopes or fossa depths. No significant relationships were found between right-left asymmetry, age, or number of teeth. With only 10% of the subjects exhibiting symmetry of the glenoid fossa depths or articular slope angles, clinicians should consider bilateral asymmetry the norm and not an anomaly. PMID- 16711270 TI - Sequence of treatment in mandibular prognathism patients. AB - Intraoral vertical ramus osteotomy (IVRO) and advancement genioplasty are effective and predictable methods of treating selective individuals with mandibular prognathism. The sequence of performing these procedures does have a clinical effect, but this is not found in the literature. The purpose of this article is to introduce the clinical implication of the sequence of procedures and to recommend the preferred sequence based on experience. A retrospective study was conducted on 75 patients treated by IVRO with or without genioplasty. In 31 patients, IVRO was performed without genioplasty; only one patient suffered from transient hypoesthesia. When IVRO was performed before genioplasty (sequence I) in 20 patients, nine suffered from mental nerve hypoesthesia. However, when IVRO was performed after genioplasty (sequence II) in 24 patients, permanent mental nerve anesthesia occurred in five patients and mental nerve hypoesthesia in 15 patients. Based on these results, it is recommended that IVRO be performed before genioplasty. PMID- 16711271 TI - Body position and jaw posture effects on supra- and infrahyoid electromyographic activity in humans. AB - The purpose of this study was to record the pattern of electromyographic (EMG) activity of supra- and infrahyoid muscles at different body positions and jaw posture tasks. The sample included 22 healthy subjects with natural dentition, bilateral molar support, and absence of posterior occlusal contacts during mandibular protrusion. Bipolar surface electrodes were located on the left supra- and infrahyoid muscles for EMG recordings. In each subject EMG activity was recorded while standing and in the lateral decubitus position, during the following jaw posture tasks: swallowing of saliva, maximal clenching in the intercuspal position, grinding from intercuspal position to protrusive edge-to edge contact position, and grinding from retruded contact position to intercuspal position. Suprahyoid EMG activity was significantly higher in the intercuspal position than in all the other jaw positions studied (mixed model with unstructured covariance matrix). Suprahyoid EMG activity was similar in both body positions studied. Infrahyoid EMG activity in the intercuspal position was significantly higher than grinding from intercuspal position to protrusive edge to-edge contact position, similar to swallowing of saliva, and significantly lower than grinding from the retruded contact position to intercuspal position. Infrahyoid EMG activity in the lateral decubitus position was significantly higher than in the standing position. The EMG pattern observed could be of clinical importance in the presence of parafunctional habits, i.e., clenching and/or grinding. The neurophysiological mechanisms involved are discussed. PMID- 16711272 TI - Headache improvement through TMD stabilization appliance and self-management therapies. AB - The purpose of this study was to assess headache response of unselected neurology clinic chronic headache patients to TMD stabilization appliance and self management therapies, and to identify features of patients whose headaches are more likely to improve from these therapies. Twenty chronic headache patients in a nontreatment control period were provided appliance and self-management therapies, evaluated five weeks after therapy, and those who chose to continue using their appliances were evaluated three months later. The mean pretreatment Headache Disability Inventory (HDI) score of 64.5 suggested the headaches were severe. After five weeks, the mean HDI score decreased by 17 percent (p<0.003), headache medication consumption dropped by 18 percent (p<0.0001), and headache symptoms decreased by 19 percent (p<0.002). Comparing the three months with pretreatment follow-up, the fourteen participants who chose to continue using their appliances had a mean HDI score decrease of 23 percent (p<0.003), headache medication consumption drop of 46 percent (p<0.001), and headache symptom decrease of 39 percent (p<0.001). There was no correlation between response and headache type (p=0.722). These results suggest appliance and self-management therapies can be beneficial for many severe headache patients, irrespective of the headache type (tension-type, migraine without aura, and migraine with aura). PMID- 16711273 TI - Anamnestic index severity and signs and symptoms of TMD. AB - The aim of this study was to determine the frequency and to characterize the symptoms and clinical signs of temporomandibular disorders (TMD) related to each severity category of Fonseca's anamnestic index in a sample of Brazilian young adults (mean age 21.61+/-1.91 years, 87% females and 13% males), by the application of an anamnestic index proposed by Fonseca (1992) and by clinical examination considering mandibular range of motion and tenderness to palpation of stomatognathic system structures. A significant number of participants were classified with mild TMD (43.2%) and moderate TMD (34.8%). Pain frequency during mastication, temporomandibular joint (TMJ) pain, and TMJ sounds were shown to be good predictors of TMD severity. Neck pain, headache, difficulty during mouth opening and lateral deviation, and tenderness to palpation of masticatory sites and during protrusion accompanied the TMD severity but failed to demonstrate differences between moderate and severe groups, showing a poor ability to determine TMD severity progression. This study suggests that not only the frequency of signs and symptoms of TMD should be determined, but also symptom severity and its relationship to the presence of clinical signs in order to discriminate patients with real treatment needs in nonpatient samples. PMID- 16711274 TI - Intra-articular and muscle symptoms and subjective relief during TMJ internal derangement treatment with maxillary anterior repositioning splint or SVED and MORA splints: A comparison with untreated control subjects. AB - Discomfort associated with wearing an intraoral splint represents a problem in the management of temporomandibular joint (TMJ) internal derangement. This study evaluated whether the use of a mandibular splint during the day and a maxillary splint at night could be more comfortable and therefore as efficacious in internal derangement treatment as a maxillary splint (AR splint). Fifty (50) patients (average age 28.8; range 14-63) with confirmed internal derangement in at least one TMJ were divided into three groups: 20 patients treated with AR splint (Group I); 20 patients treated with a SVED (Sagittal Vertical Extrusion Device) and a MORA (Mandibular Anterior Repositioning Splint) (Group II); and 10 patients who underwent no treatment (Control Group). Joint noise, pain intensity and its character (as constant or chewing/biting pain), muscular pain, and subjective relief were evaluated monthly before treatment began (T0) and for six months thereafter. The following results were found: 1. Subjects in Group I and Group II displayed a significant decrease in joint pain (p<0.001), constant pain (p<0.001), chewing/biting pain (p<0.001), joint noise and muscle pain from the beginning through the sixth month follow-ups; 2. At T1 and T2, subjects in Group II reported significantly lower discomfort associated with the devices than subjects in Group I. The use of two splints seems to be as efficacious as the use of an AR maxillary splint; however an AR splint is considered more comfortable by patients, especially during the first months of therapy. PMID- 16711275 TI - Response of temporomandibular joint intermittent closed lock to different treatment modalities: A multicenter survey. AB - This study investigated the clinical picture and different treatment methods and results at a number of institutions with the aim of establishing an effective method of treatment for intermittent closed lock (intermittent lock) of the temporomandibular joint (TMJ). The subjects were 104 patients (29 males, 65 females) diagnosed with intermittent lock among 1787 temporomandibular disorder patients. The cases were classified into two types based on the time and occasion when the intermittent lock occurred. The sudden onset type developed in 69.2%, and the habitually occurring type in 29.8%. The most common treatment was disk repositioning exercises alone (in 41 cases) followed by stabilization splints during sleeping. The highest efficacy rate (60.0%) was obtained with the combination of disk repositioning exercise and a repositioning splint followed by a rate of 52.6% with stabilization splints and 41.7% with disk repositioning exercise alone. PMID- 16711277 TI - Occlusal splints from the beginning to the present. PMID- 16711276 TI - Bilateral bifid mandibular condyle: case report and literature review. AB - Bifid mandibular condyle is an infrequent and normally asymptomatic morphological alteration of the mandibular condyle. Although the underlying cause is not clear, a number of theories have been proposed, including teratogenic effects in the embryo, vascular alterations during condyle development, and condylar remodeling following fracture. Since Schier first described this anomaly in 1948 in live individuals, further cases have been documented in the literature. We present a new case of bilateral bifid condyle. The disorder was asymptomatic and constituted a casual finding in a young male presenting for the surgical extraction of two impacted molars. PMID- 16711278 TI - Nurses cautious about donation rules. PMID- 16711279 TI - We need to re-evaluate the priorities and limits of the NHS. PMID- 16711280 TI - Do you judge your patients? PMID- 16711281 TI - Taking a stand against falls. Interview by Emma Vere_Jones. PMID- 16711282 TI - NICE guidelines to improve TB management and prevention. AB - In the UK although the majority of the population is at very low risk of TB due to vaccination, the number of cases reported is increasing. In response NICE has released fresh guidance on prevention and treatment. Nurses can play a crucial role in adherence due to the extent of their patient contact. PMID- 16711283 TI - Cardiac pacing. PMID- 16711284 TI - Obtaining a catheter specimen of urine. PMID- 16711285 TI - Supporting anxious parents in search of Internet information. AB - Parental uncertainty and anxiety is easily transmitted to sick children and can impede their recovery. Providing information to parents can help them manage anxiety. The National Service Framework for Children, Young People and Maternity Services (DH, 2003) highlights the importance of sharing information with children and parents. The internet is increasingly important as an information resource for nurses in a range of settings. This article explores how nurses can use the Internet to support anxious parents. PMID- 16711286 TI - Oxygen. PMID- 16711287 TI - A project to promote better communication with patients. AB - Good communication skills are essential when interacting with patients, especially when breaking bad news and dealing with difficult situations. National and local recognition of gaps in nurse education in such skills within the Forth Valley Acute Operating Division led to a multidisciplinary project that aimed to improve them. This article describes how this project was piloted. PMID- 16711288 TI - Nurses' smoking behaviour related to cessation practice. AB - AIM: To examine the smoking behaviour, knowledge and attitudes of nurses, their willingness to provide smoking cessation support to patients, the accessibility of training in this area and their willingness to undertake future training in this area. METHOD: A randomised sample of qualified nurses (n = 1,074) in statutory, private and voluntary sectors and across a variety of specialties were surveyed by postal questionnaire. Four focus groups were conducted in various settings before and after the survey. RESULTS: Of those who took part in the survey, 55% had never smoked, 19% were ex-smokers and 26% were smokers. Most agreed that nurses have a responsibility to help those who want to quit smoking. However, nurses who smoked rated their ability to help patients and their effectiveness as a role model lower than nurses who were ex-smokers or non smokers. CONCLUSION: Smoking prevalence among nurses is no greater than in the general female population. Nurses who smoke are less motivated to provide cessation support for patients, have less positive attitudes to the value of smoking cessation, are less likely to have received smoking cessation training and are less likely to want further training. These results have implications for nurses' own smoking status, as well as their attitudes to cessation training, health promotion practice and future research. PMID- 16711289 TI - Paruresis: a secret phobia. PMID- 16711290 TI - Selecting urinals for male patients. AB - Julie Vickerman describes some of the male urinals currently available, and highlights how appropriate assessment and provision of products can improve the quality of life for male patients with continence problems. PMID- 16711291 TI - Multiple sclerosis, body image and sexuality. AB - Janet Hutchinson explores the concept of body image and sexuality in relation to a female client with bladder dysfunction and multiple sclerosis. She also discusses the psychological and social problems that may arise from these problems. PMID- 16711292 TI - A continence pathway for acute stroke care. AB - Clare Gordon and Catherine Weller describe the development of an acute phase continence assessment tool that is to be completed within 24 hours of a patient's admission following a stroke. PMID- 16711293 TI - Urinary incontinence in nulliparous women. AB - Linda Birch and colleagues review the evidence associated with various demographic and behavioural risk factors for the development of urinary incontinence in nulliparous, pre-menopausal women. PMID- 16711294 TI - Coupling sexual health with family planning. Interview by Jaida Miah. PMID- 16711295 TI - "Until the sun of science...the true Apollo of medicine has risen": collective investigation in Britain and America, 1880-1910. PMID- 16711296 TI - Watching paint dry: organic solvent syndrome in late-twentieth-century Britain. PMID- 16711297 TI - Norwegian general hospitals, 1970-2002: county ownership--an interlude between welfare localism and state direction. PMID- 16711298 TI - Fighting venereal diseases: Scandinavian legislation c.1800 to c.1950. PMID- 16711299 TI - Thomas Vicary and the anatomie of mans body. PMID- 16711301 TI - Osteoporosis: we are neglecting our own. PMID- 16711302 TI - Augmentation procedures for osteoporotic vertebral fractures--an ongoing experiment or emerging standard of care? PMID- 16711303 TI - Thrombocytopenia. PMID- 16711304 TI - Establishing causality from case reports: was ephedra to blame? PMID- 16711305 TI - Management of acute duodenal peptic perforations. PMID- 16711306 TI - Pain outcome and vertebral body height restoration in patients undergoing kyphoplasty. AB - INTRODUCTION: Kyphoplasty, a minimally invasive technique, has recently been developed to provide immediate pain relief, biomechanical stabilization, prevention of fracture progression, vertebral height restoration, and prevention or reversal of kyphosis to patients with osteoporotic vertebral compression fractures (VCF). MATERIALS AND METHODS: We retrospectively reviewed 24 patients treated with kyphoplasty. A total of 37 vertebral levels were augmented. Visual analog scale (VAS) scores were documented in the immediate pre- and postoperative period, as well as 4, 12, and 72 weeks after the procedure. Vertebral body height restoration was assessed on postoperative x-rays. RESULTS: Mean preoperative VAS score was 9.3 and improved to 5.4 in the immediate postoperative period. At 4, 12 and 72 weeks post-operatively, mean VAS scores were 5.1, 5.9, and 6.1 respectively. All patients returned to their daily activities within 24 hours. No significant restoration of vertebral body height was observed. CONCLUSION: In regards to pain relief and postoperative functional outcome, kyphoplasty is a safe and effective treatment modality for osteoporotic VCFs, even when no significant restoration of vertebral body height is achieved. PMID- 16711307 TI - Primary care house staff attitudes toward osteoporosis management. AB - OBJECTIVE: This study assessed possible institutional and patient-related factors influencing the delivery of postmenopausal osteoporosis (PMO) care and the diagnostic priority placed on addressing PMO, relative to other common medical conditions, by primary care house staff at our institution. METHODS: A questionnaire was designed and distributed to eligible house staff at our institution. RESULTS: Approximately 50% (n = 52) of the house staff participated. The supervising clinic attending, patients' lack of insurance, accessibility to medical care, comorbid conditions, and university formulary were reported to influence decisions regarding osteoporosis care. Osteoporosis was ranked 6th of 7 medical issues (hypertension, coronary artery disease, diabetes, hypercholesterolemia, adult immunizations, osteoporosis, thyroid disease) to address during a comprehensive medical visit. CONCLUSIONS: Our institution's primary care house staff reported multiple influences on decision making regarding osteoporosis care, and an overall low priority to address this issue. Based on PMO's associated morbidity and mortality, primary care training programs are challenged to put resources toward optimizing house staff delivery of osteoporosis care. PMID- 16711308 TI - Omental plugging for large-sized duodenal peptic perforations: A prospective randomized study of 100 patients. AB - BACKGROUND: Due to friable margins and the moribund state of the patient, managing giant duodenal perforations (>20 mm in diameter) is a challenging task. METHODS: A prospective randomized study of 100 patients with large-sized (> 20 mm) duodenal peptic perforation comparing omental plugging (study group) with omentopexy (control group) was carried out. RESULTS: Size of the perforation varied between 20 to 30 mm. No study group patients developed a postoperative perforation site leak, as compared with 6 patients in the control group. Gastric outlet obstruction was significantly less at 6 weeks and 5 years in the study group as compared with the control group, and mortality was significantly less in the study group. CONCLUSION: It was concluded that omental plugging was a safe and reliable method of treatment for large-sized duodenal peptic perforations. PMID- 16711309 TI - Factors associated with delayed initiation of HIV medical care among infected persons attending a southern HIV/AIDS clinic. AB - BACKGROUND: Despite the proven benefits conferred by early human immunodeficiency virus (HIV) diagnosis and presentation to care, delays in HIV medical care are common; these delays are not fully understood, especially in the southern United States. METHODS: We evaluated the extent of, and characteristics associated with, delayed presentation to HIV care among 1,209 patients at an HIV/AIDS Outpatient Clinic in Birmingham, Alabama between 1996 and 2005. RESULTS: Two out of five (41.2%) patients first engaged care only after they had progressed to CDC-defined AIDS. Among these, 53.6% were diagnosed with HIV in the year preceding entry to care. Recent presentation (2002 - 2005), male sex, age > or = 25, Medicare or Medicaid insurance coverage, and presentation within six months of HIV diagnosis were independently associated with initiating care after progression to AIDS. CONCLUSIONS: A high proportion of patients entered clinical care after experiencing substantial disease progression. Interventions that effectively improve the timing of HIV diagnosis and presentation to care are needed. PMID- 16711310 TI - Sleep disturbance in patients with lichen simplex chronicus and its relationship to nocturnal scratching: A case control study. AB - BACKGROUND: Lichen simplex chronicus (LSC) is a common pruritic disorder resulting from repeated rubbing and scratching. Nighttime pruritus is a common feature in LSC and may disrupt the sleep pattern. The aim of this study is to determine whether there are sleep abnormalities in patients with LSC. PATIENTS AND METHODS: Fifteen patients with LSC and 15 age-, sex- and body mass index matched control subjects were enrolled in the study. No participant had any other medical or psychiatric illness. All subjects were evaluated by overnight polysomnography, scratch electrodes, Epworth sleepiness scale and a general questionnaire for demographic data and sleep problems. RESULTS: There were no significant differences in the groups for total sleep time, sleep efficiency, sleep latency, rapid eye movement (REM) latency, percentage of stage 1 non-REM sleep and REM sleep. The percentage of stage 2 non-REM sleep was higher (P < 0.05) and the percentage of slow wave sleep (stages 3 and 4) was lower in the study group (P < 0.05) than in the controls. The patient group had a mean of 15.9 +/- 7.5 arousal index and 22.8 +/- 14.1 awakenings compared with 9.5 +/- 3.1 and 10.4 +/- 3.9, respectively, in the controls (P < 0.05, P < 0.01, respectively). Arousals of patients were mainly observed in non-REM sleep. The number of scratching bouts ranged from 6 to 20 per night. Scratching episodes were observed frequently during stage 2 non-REM sleep. CONCLUSION: Polysomnographic findings of patients with LSC demonstrated that sleep structure is disturbed by arousals and awakenings related to scratching bouts during sleep. PMID- 16711311 TI - Follow-up study of medication errors reported to the vaccine adverse event reporting system (VAERS). AB - BACKGROUND: A study was done to determine if the apparent medication errors found in the Vaccine Adverse Event Reporting System (VAERS) database are true errors, and if true errors are found, to determine what corrective action was taken. Furthermore, if a true error did not occur, we wanted to determine at what point the misinformation was entered into the system. METHODS: The VAERS database was searched for reports received between July 1, 2001 and June 30, 2002 which had either been classified as "error" or the word "error" appeared in the text of the report. The database was also searched for reports which indicated that the measles-mumps-rubella (MMR), diphtheria-tetanus-pertussis (DTP), diphtheria tetanus-acellular pertussis (DTaP) or diphtheria-tetanus-pertussis-haemophilus (DTPH) vaccinations had been administered at an age outside of the usual recommendation. RESULTS: A total of 119 reports of possible errors were found. Follow-up was successful in 102 (86%) cases. Additional information obtained showed that 26 cases were actual medication errors. Seventy-six cases were not actual medication errors; 9 cases were physician decisions, 37 cases were data entry errors and 30 cases were reporter errors. CONCLUSION: The nature of the actual errors was similar to those reported previously; wrong inoculum, improper interval, wrong route of administration, and overdose. Many errors could have been prevented by more attention to detail. Remedial action usually consisted of retraining. The new requirement that all medications be barcoded, purchasing products from different manufacturers and segregation of vials may help prevent vial confusion. PMID- 16711312 TI - Thrombocytopenia in adults: A practical approach to evaluation and management. AB - With the widespread use of automated cell counters, clinicians in any field of medicine may encounter thrombocytopenia. The symptomatology may vary greatly and the underlying cause may be either inconsequential (pseudothrombocytopenia) or life threatening. It is important to be aware of common conditions leading to thrombocytopenia and have a systematic approach to evaluation and management of these patients. In this review we highlight common etiologies seen in adult patients with thrombocytopenia. A brief description and management approach to common conditions, as well as to conditions that may be less frequent but require emergent intervention, is provided. Often the clinician is called upon to make a decision about platelet transfusions. The review also discusses the various types of platelet products available for transfusion and gives guidance regarding selection of the appropriate product, indications and contraindications, and suggested target platelet counts for various clinical situations. PMID- 16711313 TI - Sudden cardiac death. AB - Sudden cardiac death (SCD) due to ventricular tachyarrhythmias is a leading cause of death in the United States. Various etiologies, including ischemic and nonischemic cardiomyopathies, hypertrophic cardiomyopathy, valvular or congenital heart diseases and other less common disorders, may result in SCD. Beta blockers are the only class of medications that have been shown to be beneficial in the primary prevention of SCD. However, recently, aldosterone antagonism early after myocardial infarction has also been shown to significantly reduce the risk of SCD. Multiple trials have elaborated on the potential benefits of implantable cardioverter defibrillators (ICD) in appropriately selected patients. However, there is still some controversy regarding the optimum period for ICD implantation, and its cost-effectiveness. An evidence-based approach to primary and secondary prevention of SCD is presented. Management of out-of-hospital cardiac arrest is briefly discussed. PMID- 16711314 TI - Transient blindness due to posterior reversible encephalopathy syndrome following ephedra overdose. AB - Posterior reversible encephalopathy syndrome (PRES), also known as reversible posterior leukoencephalopathy syndrome (RPLS), is most often associated with hypertensive emergencies and is characterized by seizures, mental status changes and visual disturbances. We report a case of a previously healthy young man who developed multiorgan failure and transient cortical blindness following ingestion of a performance-enhancing ephedra-based supplement. Neuroimaging findings confirmed the clinical suspicion of PRES. Radiographic abnormalities and neurologic dysfunction subsequently resolved with correction of his systolic blood pressure. This case emphasizes the need for prompt treatment and consideration of toxic ingestions in patients presenting with hypertension related end-organ dysfunction. PMID- 16711315 TI - Cardiogenic shock in hypothyroidism. AB - A patient with minimal coronary artery disease presented in cardiogenic shock when her previously undiagnosed hypothyroid state was complicated by an episode of AV nodal re-entrant tachycardia. She did not respond to multiple pressors, and recovered dramatically after starting thyroid supplementation. Hypothyroidism caused her lack of responsiveness to pressors and perpetuated her hypotension and increased filling pressures long after she reverted to a sinus rhythm. Our case dramatically demonstrates the severe lack of physiologic reserve that can be associated with hypothyroidism. PMID- 16711316 TI - Significant absorption of oral vancomycin in a patient with clostridium difficile colitis and normal renal function. AB - Orally-administered vancomycin is poorly absorbed in most patients, usually producing minimal or subtherapeutic serum concentrations. Bowel inflammation may enhance absorption of oral vancomycin, particularly in those with renal failure. A 77-year-old female with Clostridium difficile (C difficile) colitis and normal renal function was treated with high doses of oral vancomycin and achieved serum concentrations in the therapeutic range. To our knowledge, this is the first report of a patient with C difficile colitis and normal renal function to develop therapeutic serum concentrations following oral administration of vancomycin. PMID- 16711317 TI - Fecal impaction and systemic inflammatory response syndrome in a young male with cerebral palsy. AB - Symptoms of fecal impaction extend from constipation, anorexia, nausea, vomiting and abdominal pain, to full blown sepsis. We present the case of a patient with cerebral palsy and mental retardation, who presented to the Emergency Department with a 3-day history of diffuse abdominal pain and fecal incontinence. Evaluation revealed severe fecal impaction. The patient developed systemic inflammatory response syndrome (SIRS), with negative workup for underlying etiology. He responded well to digital disimpaction and antibiotics. Our case illustrates the serious sequelae of fecal impaction, which should be considered in patients with neurologic disorders and SIRS. PMID- 16711318 TI - Massive fecal impaction presenting with megarectum and perforation of a stercoral ulcer at the rectosigmoid junction. AB - A 25-year-old male with lifelong constipation presented to the emergency department with an acute abdomen. Initial resuscitation was performed, and the patient underwent urgent laparotomy. He was found to have feculent peritonitis with megabowel involving the rectum and sigmoid colon and a stercoral ulcer with full thickness erosion, and perforation was also identified on the anti-mesocolic surface at the rectosigmoid junction. Abdominal irrigation and subtotal colectomy with proximal fecal diversion was performed. This case illustrates that recognition of severe, chronic constipation should lead to interventions including disimpaction and aggressive medical management. When indicated, megabowel can be managed surgically in an elective setting based on anatomic findings and physiologic studies. Peritonitis is an ominous late finding in patients with severe constipation. PMID- 16711319 TI - Acute bowel ischemia following spinal surgery. AB - Acute mesenteric ischemia is a morbid condition that may be difficult to diagnose due to nonspecific nature of its symptoms. To our knowledge, such a complication has not previously been reported after spinal surgery via the posterior approach. We describe the case of a 43-year-old woman who developed acute mesenteric ischemia several days after a surgical procedure for a lumbar spondylolisthesis via the posterior route. This chronic course is suggestive for venous intestinal ischemia. Prone position and hypotension during the procedure may have favored blood stasis and mesenteric vein occlusion in this patient with an inherited hypercoagulable state. PMID- 16711320 TI - Anterior uveitis, inflammatory bowel disease, and ankylosing spondylitis in a HLA B27-positive woman. AB - A woman developed anterior uveitis at age 24, inflammatory bowel disease at age 29, and ankylosing spondylitis at age 45 by history. There were frequent recurrences. An HLA-B27 test was positive at age 53. The literature indicates that all of these conditions together in a HLA-B27-positive woman are uncommon. Physicians should be alert to the possibility that a patient might develop another of these associated diseases years after presentation of the first condition and educate their patients accordingly. PMID- 16711321 TI - Chest and abdominal injuries caused by seat belt wearing. AB - The authors report an original case of seat belt syndrome. Sternal fractures are common in patients with seat belt injuries. Its association with blunt bowel trauma is rarely related in the literature. Distracted injury has contributed to delay the diagnosis of intestine injury. The presence of a seat belt sign must lead to the consideration of occult injuries especially abdominal. Close observation and serial examination should be the rule. PMID- 16711322 TI - Flail chest in a neonate resulting from nonaccidental trauma. AB - The authors present a 21-day-old infant who sustained a flail chest as a result of nonaccidental trauma. Initial treatment included endotracheal intubation and mechanical ventilation for hypoxemic respiratory failure followed by the administration of continuous positive airway pressure by nasal cannula. Further evaluation resulted in the identification of nonaccidental as the mechanism of injury. The pathophysiology of flail chest, its etiology, and treatment options are reviewed. In the absence of a documented history of significant thoracic injury or the presence of metabolic bone disease, nonaccidental trauma is the most likely diagnosis in infants and children with a flail chest. PMID- 16711324 TI - Epstein Barr virus hepatitis: case series and review. AB - Epstein Barr virus (EBV) infection causes asymptomatic liver-associated enzyme abnormalities in 80 to 90% of cases which are often unrecognized. Patients with acute EBV infections may also develop cholestatic hepatitis with associated jaundice and hepatitis with moderate elevations in the transaminase levels. Other gastrointestinal complications associated with EBV may include splenic rupture, liver failure due to acute and/or chronic EBV infection, and perhaps, autoimmune hepatitis and hepatocellular carcinoma. This article presents a case series of EBV infections with clinically significant hepatitis and reviews the literature on the gastrointestinal complications of EBV. PMID- 16711323 TI - Incidental finding of congenital thoracic malformations in adult population. AB - OBJECTIVE: Evaluation of abnormal adult chest computed tomography (CT) scans. STUDY DESIGN: Retrospective series of 3 cases. SETTING: Two University-based hospitals. INTERVENTION: Three adult patients (age range 56 to 61) underwent chest CT scans. Two were trauma patients and one complained of chest pain with a negative cardiac workup. One CT scan revealed a lung mass that was highly suspicious for malignancy and the other two CT scans were suspicious for diaphragmatic hernia. Two patients underwent elective surgery and the third underwent emergent surgery. RESULTS: The CT scan of patient 1 was suspicious for malignancy; however, the pathology of the lung mass revealed only inflammation consistent with an intralobular bronchopulmonary sequestration. Patient 2 had a congenital Morgagni hernia with omentum and colon in the mediastinum, and patient 3 had a diaphragmatic defect combined with a bifid sternum and defective pericardium, fulfilling three criteria of pentalogy of Cantrell. CONCLUSION: Thoracic congenital malformations may go unnoticed and unsuspected until adulthood. They should be kept in mind when reviewing unusual CT scans of the chest in adults. Their surgical management, however, is similar to their counterparts in infancy. PMID- 16711325 TI - A case of occult oropharyngeal dysphagia. PMID- 16711326 TI - Talc pleurodesis for recurrent pleural effusions: is cure worse than the actual insult? PMID- 16711328 TI - Hospitalist physicians as educators in a community hospital: the trainee's view. PMID- 16711327 TI - Unilateral pulmonary edema secondary to idiopathic rupture of mitral chordaea tendinae. PMID- 16711329 TI - Normal chest radiograph in terminal respiratory failure due to amyotrophic lateral sclerosis. PMID- 16711330 TI - Patient's page. May is National Osteoporosis Awareness and Prevention Month. PMID- 16711331 TI - My perspective on the history of emergency medicine. PMID- 16711332 TI - Childhood lymphoma and leukaemia. PMID- 16711333 TI - Colorectal carcinoma, preoperative evaluation by spiral computed tomography. AB - OBJECTIVES: To assess the capability of spiral computed tomography (CT) scan in preoperative evaluation of colorectal carcinoma (CRC). METHODS: A cross sectional study on 52 patients with recent histopathologic diagnosis of CRC was conducted over a period of one year at the Aga Khan University Hospital, Karachi. All these patients underwent spiral CT with oral and intravenous contrast administration. Surgery was subsequently done in all cases and surgical specimens were sent for detailed pathologic analysis. The radiologic findings on the CT scans were compared with the pathological findings. RESULTS: The results proved that spiral CT had 60% sensitivity and 83% specificity for assessment of local spread of disease, 66% sensitivity and 76% specificity for the evaluation of lymph nodal metastases and 89% sensitivity and 94% specificity for hepatic metastases. In all the cases, the visualized tumour growth with wide zone of resection and regional nodal chains were surgically removed. It was however, the distant metastases which made a difference to the type of curative or palliative surgery planned and in this study spiral CT had 92% accuracy for detection of hepatic metastases. CONCLUSION: With technological advances and improvement in imaging protocols the results for local tumor spread are expected to improve, however based on the accuracy in detecting hepatic metastases in clinically unsuspected patients, this study proves that spiral CT has a significant role in preoperative evaluation and subsequent management of CRC. PMID- 16711334 TI - Sentinel lymph node scintigraphy and radioguided dissection in breast carcinoma: an initial experience at Aga Khan University Hospital. AB - OBJECTIVE: To evaluate the efficacy of SLN biopsy using imaging and gamma probe in breast cancer and to establish this technique at Aga Khan University Hospital. METHODS: Thirty two patients (mean age 33-76 yrs) with operable breast carcinoma (4 with post-neo adjuvant therapy) with clinically negative axilla were studied. In 28 patients simultaneous axillary dissection was performed. Sentinel lymph nodes (SLN) scintigraphy was performed a day before surgery by injecting Tc-99m labeled nannocolloid sub-dermally in the peri-areolar region in 24 and peri tumoral in remaining 8. First lymph node (LN) to appear to on the scan was labeled as SLN and marked on the skin. Blue dye was also injected in all patients and blue and hot LN was explored in the axilla using gamma probe. RESULTS: The sentinel LN was identified in 31 patients (96.9% success rate) while in one patient (3.1%) SLN was not visualized on the scan (negative study) and this was a post-neo-adjuvant therapy case. The blue dye successfully localized the sentinel LNs in all 32 cases The gamma probe guided localization was successful in all 31 while in one case with post-neo-adjuvant chemotherapy it failed. In 9 out of 32 (28.1%) cases SLN was positive for metastasis and in 6 out of these 9 (66.6%) the SLN was the only metastatic node. This includes the post-neo-adjuvant case as well. There was no case of skip metastasis, i.e. negative SNL and positive other axillary nodes. In remaining 23 (71.9%) cases SLN was negative for metastasis and in all, axilla was free of disease (NPV of 100%). CONCLUSION: Lymphoscintigraphy with gamma probe guided SNL biopsy is safe, simple and highly reliable technique. With blue dye technique it reduces the blindness of the procedure if performed independently but increases the cost. Finally, this can accurately stage the axilla, possibly allowing axillary dissection to be foregone in patients where the SLN is clear. PMID- 16711335 TI - She may reach a facility but will still die! An analysis of quality of public sector maternal health services, District Multan, Pakistan. AB - OBJECTIVE: To assess the availability, utilization and functional capacity of the public health delivery system in terms of human resources, equipment, supplies and health provider knowledge regarding management of obstetric complications. METHODS: A cross-sectional study was conducted in March 2003 for Multan's public health delivery system. A complete inventory of equipment, supplies, drugs, staff availability and knowledge of obstetric complications management were assessed. RESULTS: The number and distribution of public sector facilities serving a population of 3.2 million was well beyond the minimum requirement for basic obstetric care but fell short for comprehensive Emergency Obstetrical Care (EmOC). Utilization patterns were skewed to the tertiary facility for normal and complicated deliveries. Partograms were not available, magnesium sulphate was only available at the tertiary facility, while basic supplies like iron and folate tablets, broad spectrum antibiotics, oxytocics, gloves and sutures were not available at primary or secondary facilities. Knowledge scores regarding management of obstetric complications were less than average, especially among nurses at basic or comprehensive EmOC facilities. Intensity of labour pains (nurses 59%) at basic EmOC facility and cervical dilatation by doctors at basic and comprehensive facilities were the most frequent responses for monitoring in labour. CONCLUSION: Strengthening the four interlinked health system elements- human resources, access to, use and quality of services is essential though not sufficient. PMID- 16711336 TI - Incidence of acute respiratory infections in children 2 months to 5 years of age in periurban communities in Karachi, Pakistan. AB - OBJECTIVE: To measure the incidence of acute respiratory infections and burden of respiratory pathogens in children aged two months to five years. METHODS: Four periurban communities in Karachi were selected for the study. The children, identified with fever and cough during community surveillance at regular intervals, were referred to especially established study clinics. These children were diagnosed to have "no pneumonia", "pneumonia" and "severe pneumonia" as per IMCI guidelines. To identify the causative organisms, children with pneumonia and severe pneumonia were investigated with oropharyngeal swabs and blood culture. RESULTS: Acute respiratory infection was seen in 5884 children during 1st February 2002 to 31st January 2003. Of these, 1097 children had pneumonia and severe pneumonia, with an incidence 440.3/1000 children per year for Acute Respiratory Infections and 82.1/1000 children per year for pneumonias. Haemophilus influenzae, Streptococcus pneumoniae and Klebsiella pneumoniae were isolated from 10.9%, 3.7% and 8.5% of oropharyngeal swabs respectively. Extrapolating from the results of this study, the total number of cases of pneumonias in children aged less than five years in Pakistan is estimated to be 213,116 per year due to H. influenzae, and 71,864 per year due to S. pneumoniae. CONCLUSION: Incidence of acute respiratory infections in children varies in different communities and is a common cause of morbidity. PMID- 16711337 TI - Role of ICT malaria immunochromatographic test for rapid diagnosis of malaria. AB - OBJECTIVE: To evaluate the sensitivity and specificity of immunochromatographic test (ICT) malaria p.f/p.v using microscopy as the gold standard diagnosis. METHODS: Five hundred and sixty patients of both sexes and all age groups with clinical suspicion of malaria were studied. Venous blood was collected for microscopy and ICT. Thick and thin films prepared and stained with Leishman's stain were examined. ICT malaria test was performed and interpreted according to manufacturer's instructions. Data was analyzed using Epi-6. RESULTS: A total of 560 cases were studied, 339 males and 221 females with age ranges between 2 to 73 years. Seventy two (12.85%) cases had parasitaemia (with or without sexual forms). On microscopy 65 (11.6%) cases had asexual-stage parasitaemia and 7 (1.25%) cases had P. falciparum gametocytes only. Thirty two cases were infected with P. falciparum, 29 with P. vivax and 4 had mixed infection. For P. falciparum the ICT was 97.0% sensitive, 98.3% specific, with positive predictive value (PPV) of 78.0% and a negative predictive value (NPV) of 99.8%. For P. vivax the sensitivity was only 89.7%, specificity 97.9%, PPV was 70.3% and NPV 99.4%. CONCLUSION: Our results are in concordance with previous studies. Rapid tests though expensive are simple to perform and effective diagnostic tools of malaria. They can be used selectively, though microscopy remains the gold standard diagnosis, economical and accurate if performed by skilled technologists. PMID- 16711338 TI - Pre-existing malnutrition and treatment outcome in children with acute lymphoblastic leukaemia. AB - OBJECTIVE: To assess the effects of pre-existing malnutrition on the treatment outcome of children with acute lymphoblastic leukaemia. METHODS: One hundred and sixty three patients with Acute Lymphoblastic Leukaemia (ALL) below the age of 14 years with L1 and L2 FAB morphology were included in this study. Treatment protocol used was FBM. Patients were classified according to Waterlow classifications of malnutrition (1976). Group-I, as Under-Nourished children (UNC) and Group-II as Well-nourished children (WNC). Percentages in both groups were calculated with respect to total expired, relapses and completed treatment. RESULTS: In Group-I (UNC) 46% completed treatment and were alive, 9.8% relapsed and 45% expired. In Group-II (WNC) 59% completed treatment and were alive, 21.3% relapsed and 19% expired. Overall, in WNC group 13.5% completed treatment and were alive, 8% relapsed and 7.3% expired. In UNC group 28.8% completed treatment and were alive, 6% relapsed and 27% expired. CONCLUSION: Pre-Existing malnutrition adversely effects the treatment outcome in children with Acute Lymphoblastic Leukaemia (ALL). PMID- 16711339 TI - Early neonatal mortality: effects of interventions on survival of low birth babies weighing 1000-2000g. AB - OBJECTIVE: In order to improve our newborn care, we instituted several changes along with training of medical and nursing staff and compared survival rates in babies < 2000g before and after these changes. We also measured Perinatal Mortality Rate (PMR) and Neonatal Mortality Rate (NMR) in general, percentage of Low Birth Weight (LBW) babies and causes of early neonatal deaths at Lady Dufferin Hospital (LDH). METHODS: It was an intervention study design. All admissions to NICU between 1998 and 2000 were entered in the register. Data included high risk obstetric factors, gestational age, birth weight, APGAR score, gender, need for resuscitation, diagnosis, complications and outcome. Data of rest of the babies was recorded from operation theatre, labour room and postnatal ward registers in a separate register. RESULTS: Of 783 perinatal deaths, 488 were stillbirths and 295 were early neonatal deaths; 2498/14867 (17%) babies were LBW. The main causes of early neonatal mortality included prematurity and related complications (35%), congenital malformations (23%), sepsis 19%), and birth anoxia (16%). Most (27/295 77%) deaths occurred in babies weighing <2500g. Deaths due to prematurity and related complications in babies weighing between 1000 1499g decreased from 17/33 (51%) in 1997 (to 13/33 (39%) 9/45 (20%), 2/38 (5%) in 1998-99 and 2000 respectively. Sepsis related deaths decreased in babies weighing between 1500 to 1999g from 5/57(8.7%) in year 1997 to 7/77 (9%), 3/76 (4%), 4/96 (4%) in 1998, 99, and 2000 respectively. The PMR/NMR decreased from 58/22.4 in year 1997 to 39/15.6, 44.4/18.6, 38.2/12.3 for year 1998, 1999 and 2000 respectively. CONCLUSION: There was a significant reduction in mortality in LBW babies after training of medical and nursing staff. Reduction in overall PMR & NMR was also due to decrease in mortality in LBW babies. PMID- 16711340 TI - The relationship of periodontal disease to cardiovascular diseases--review of literature. AB - Association of Oral and Systemic diseases has gained importance because the high occurrence of oral diseases is an extremely common source of infection. Epidemiological Studies have presented periodontal diseases as a risk factor for development of cardiovascular diseases. A chronic oral infection such as periodontitis is a constant potential source of infection and has now been considered as a separate risk factor for cardiovascular diseases, cerebrovascular diseases, peripheral arterial disease and respiratory diseases as well as delivery of low-birth-weight infants. The possible pathways linking oral infections to systemic diseases are metastatic infections, bacterial endotoxins, and systemic vascular injury. People with a history of periodontal disease and/or tooth loss were found at higher risk for Peripheral arterial disease (PAD) as compared to those without periodontal disease and/or tooth loss. All studies on the relationship of periodontal diseases to cardiovascular diseases are inconclusive and most of the data is based on epidemiological studies. PMID- 16711341 TI - Surgery for prolactinomas. AB - A large proportion of prolactin secreting tumours of the pituitary gland are treatable by dopamine agonist drugs. However certain subgroups of these patients are better managed by surgical excision. The indications for surgery in prolactinomas remain controversial and this paper attempts to clarify these with illustrative cases. The factors favouring a surgical approach include poor tolerance of long term medical therapy, progression of visual deficits or recurrence while on medical treatment, cystic change in the tumour with pressure effects, CSF fistula secondary to tumour shrinkage and prophylactic surgery in tumours anticipated to cause CSF leak following medical therapy. The size and invasiveness of the tumour and the prolactin level are also important determinants of treatment planning. The indications for surgery in prolactinomas are discussed and illustrative cases are presented. PMID- 16711343 TI - Cerebral embolism: distal subclavian disease as a rare etiology. AB - Subclavian artery occlusive disease is usually secondary to persistent compression caused by thoracic outlet syndrome (TOS) and rarely due to focal atherosclerosis. Emboli from diseased vessel can flow retrograde to the vertebral or carotid arteries to produce ischemic infarct with or without neurological deficit. We are reporting two cases of distal subclavian artery disease presenting with cerebral embolization, an unusual manifestation. Such surgically correctable lesions producing cerebral emboli and stroke needs consideration while evaluating patients with unusual presentation to prevent further occurrence of stroke. PMID- 16711342 TI - Synchronous occurrence of gastrointestinal stromal tumor and gastric adenocarcinoma: a case report. AB - Only a few cases of synchronous occurrence of gastric adenocarcinoma and gastrointestinal stromal tumour are reported in literature. We report a case of 70 years old lady who presented with symptoms of nausea, vomiting, epigastric pain and weight loss. Barium meal examination showed stricture of the body and antrum. Gastric endoscopic biopsy showed poorly differentiated adenocarcinoma. Total gastrectomy revealed a 2cm submucosal nodule of low risk gastrointestinal stromal tumour along with poorly differentiated adenocarcinoma. Patient died 18 months later. PMID- 16711344 TI - Primary mediastinal germ cell tumor--successful curative resection following chemotherapy--a case report. AB - Primary mediastinal germ cell tumours are relatively rare and account for only a small portion of all the mediastinal tumours. We present a case of a primary mediastinal germ cell tumour in a 14 years old Afghani boy. PMID- 16711345 TI - Psoas abscess secondary to Pott's disease--an unusual presentation in a young child. AB - Psoas abscess in neonates and infants are rare. Primary psoas abscesses are said to be more common in young children. Limping, fever and abdominal pain has been described to be the way psoas abscesses usually present. The authors describe the unusual presentation and successful treatment of a young child with a unilateral psoas abscess secondary to advanced spondylodiscitis. PMID- 16711346 TI - Living with globalization: a menace or a chance? PMID- 16711347 TI - Correlates of being diagnosed with depression or anxiety problems in under 18 year olds--United States. PMID- 16711348 TI - How to kill synthetic biology. PMID- 16711349 TI - The end of the Everglades? PMID- 16711350 TI - Disruptive threats. PMID- 16711352 TI - Hard landscape. PMID- 16711351 TI - Trace elements. PMID- 16711353 TI - NASA's reverse thrust. PMID- 16711355 TI - The flipping point. PMID- 16711354 TI - Body blazes. PMID- 16711356 TI - The new geopolitics. PMID- 16711358 TI - The secrets of supervolcanoes. PMID- 16711357 TI - The implicit prejudice. PMID- 16711359 TI - Engineering life: building a fab for biology. PMID- 16711360 TI - Wading in waste. PMID- 16711362 TI - Dependable software by design. PMID- 16711361 TI - Toward better pain control. PMID- 16711363 TI - A new assault on HIV. PMID- 16711364 TI - The science behind Sudoku. PMID- 16711365 TI - Carbon hooch. PMID- 16711366 TI - The ultimate blood test. PMID- 16711367 TI - What are the physical and chemical changes that occur in fireworks? PMID- 16711368 TI - How do antibiotics kill bacterial cells but not human cells? PMID- 16711369 TI - [The use of biomarkers in cancer prevention]. AB - Biomarkers in cancer prevention are increasingly important tools in primary prevention and in intervention by chemopreventive agents. Biomarkers can be utilized as indicators of exposures, effects and individual susceptibility to cancer. Sampling of biomarkers in relation to exposure may have a great impact on the reliability of mechanism of action. The recent developments in genomics provides us with an opportunity to investigate simultaneously numerous oncogenes, tumor-suppressor genes, phenotypic changes in proteins and to utilize the same samples for better understanding of possible ways to disrupt carcinogenesis. Identification of high cancer risk biomarkers is possible also by traditional molecular genetics techniques. Chromosomal studies are based on peripheral blood lymphocyte (PBL) cultures, which may present mutation as a cytogenetic change on chromosome structures. International databases are available for evaluation of high cancer risk among those people who are carrying high incidence of chromosomal aberrations. The biomarkers are suitable to indicate the need for intervention in high risk groups. In those cases, where removal of environmental hazards are not efficient enough in preventive measures since genetic damages are irreversible, it may be possible to introduce chemoprevention at the same time, which includes changes in lifestyle, smoking and drinking habits as well as prescription of vitamins, antioxidants, minerals etc. This paper summarizes the current knowledge on biomarkers indicative of high risk groups and/or individual susceptibility measured by different methodologies, mainly based on Hungarian human genotoxic monitoring studies, which may help to understand the use of biomarkers in everyday medical practice. PMID- 16711370 TI - [New trend in treatment in patients with chronic hepatitis C]. AB - The combination of pegilalt interferon plus ribavirin is significantly more effective than conventional interferon plus ribavirin, and is thus the current therapy of choice for patients with chronic hepatitis C. Forty-eight weeks of treatment with combination of pegilalt interferon plus ribavirin has produced overall sustained virologic response 54-56% in patients with chronic hepatitis C. This article summarize the new therapy of chronic hepatitis C, the antiviral therapy of hepatitis C virus patients with normal aminotransferase levels, the therapy in patients with compensated cirrhosis and who were nonresponders to previous combination therapy, the effect of antiviral therapy on necroinflammation and fibrosis, and the favourable effect of haemopoetic growth factors on results of antiviral therapy. A substantial proportion of patients with chronic hepatitis C have persistently normal alanin-aminotransferase levels. Many studies have indeed provided evidence that although the majority of hepatitis C virus patients with normal alanin-aminotransferase may have active and progressive liver disease. Initiation of antiviral therapy might be decided mainly on the basis of histological findings. Extending the treatment duration from 48 to 72 weeks in patients with hepatitis C virus genotype 1 significantly reduces relapse rates and increases sustained virological rates. Induction phase with a higher dose of pegilalt interferon may be of value in improving outcomes in patients with genotype 1 who were nonresponders to previous combination therapy. The combination of pegilalt interferon and ribavirin was effective and well tolerated in patients with compensated cirrhosis. Interferon-based therapy may reduce hepatic inflammation in the absence of a sustained virologic response. This results suggest that interferon maintenance therapy may slow disease progression. The virologic response rates could be improved by utilizing haemopoetic growth factors, as opposed to reducing the ribavirin and interferon dose. PMID- 16711372 TI - [Informed consent: a pragmatic view]. AB - The overall goal of this study is a short introduction into the Hungarian legal basis, and practical circumstances of informed consent trough the cases of the Data Protection Commissioner of Hungary, the Supreme Court of Hungary, and the Constitutional Court. The author begins by describing the basic law with an introduction to the dogmatic background of informed consent, as a common law category. [corrected]. PMID- 16711371 TI - [Selective bowel decontamination]. AB - Infective complications play major role in mortality of high risk patients demanding intensive care. Selective Bowel Decontamination prevents endogenous infections by reducing the number of potentially pathogen microbes (aerobic bacteria, fungi) in the oropharynx and gastrointestinal tract, saving anaerobic bacteria. It had been used 20 years ago for the first time. Authors survey it's literature ever since. Selective Bowel Decontamination is performed by the mixture of antibiotics and antimycotic drug, administered orally in hydrogel, and suspension form in nasojejunal tube. The number of Gram negative optional aerobic bacteria and fungi decrease significantly in the gut, and the microbial translocation is following this tendency. Foreign authors achieved good results in acute necrotizing pancreatitis, after liver transplant, in polytrauma, in serious burn and in haematological malignancies. According to the literature Selective Bowel Decontamination shows advantages in selected groups of high risk surgical patients. In some studies the administration took few months, but the minimum time was one week. There was no report of increasing MRSA appearance. Regular bacteriological sampling is highly recommended in order to recognize any new antibiotic resistance in time. PMID- 16711373 TI - [Deep freezing of oocytes for preserving the fertility of young female oncology patients treated with aggressive radio- and/or chemotherapy]. AB - The aggressive radiotherapy and chemotherapy used for treatment of oncological patients--through the damage of germ cells--may cause decrease of fertility or complete infertility. There are relatively few effective clinical options for preserving female fertility. The results of clinical experiments connected with ovarian tissue cryopreservation and oocyte freezing are very promising and indicate that we may have tools for female fertility preservation in the future. In this paper, 1) the future role of oocyte cryopreservation in fertility preservation of female oncological patients, and 2) the authors' preliminary results with oocyte cryopreservation are summarized. PMID- 16711374 TI - [The Marikovszkies in the history of the Hungarian medicine]. PMID- 16711375 TI - [Remembering Istvan Kulcsar (1901-1986)]. PMID- 16711376 TI - Infant mortality statistics from the 2003 period linked birth/infant death data set. AB - OBJECTIVES: This report presents 2003 period infant mortality statistics from the linked birth/infant death data file by a variety of maternal and infant characteristics. The linked file differs from the mortality file, which is based entirely on death certificate data. METHODS: Descriptive tabulations of data are presented and interpreted. Excluding rates by cause of death, the infant mortality rate is now published with two decimal places. RESULTS: The U.S. infant mortality rate was 6.84 infant deaths per 1,000 live births in 2003, a return to the rate in 2001, compared with 6.95 in 2002. Infant mortality rates ranged from 4.83 per 1,000 live births for Asian or Pacific Islander mothers to 13.60 for non Hispanic black mothers. Among Hispanics, rates ranged from 4.57 for Cuban mothers to 8.18 for Puerto Rican mothers. Infant mortality rates were higher for those infants whose mothers were born in the 50 States and the District of Columbia, were unmarried, or smoked during pregnancy. Infant mortality was also higher for male infants, multiple births, and infants born preterm or at low birthweight. Infants born at the lowest birthweights and gestational ages have a large impact on overall U.S. infant mortality. Nearly one-half (49 percent) of all infant deaths in the U.S. in 2003 occurred to the 0.8 percent of infants whose birthweight was less than 1,000 grams. The three leading causes of infant death- Congenital malformations, low birthweight, and SIDS--taken together accounted for 45 percent of all infant deaths. For infants of non-Hispanic black mothers, the cause-specific infant mortality rate for low birthweight was nearly four times that for infants of non-Hispanic white mothers. For infants of non-Hispanic black and American Indian mothers, the SIDS rates were more than double the rate for non-Hispanic white mothers. PMID- 16711377 TI - Responding to a trial subpoena. PMID- 16711378 TI - Surfing the Web on company time. PMID- 16711379 TI - The basics of due diligence for buyers. PMID- 16711380 TI - Fraud in dentistry. PMID- 16711381 TI - Toni Talbot's top 10 tips to keep your best employees. PMID- 16711382 TI - Consider giving the "gift of life". PMID- 16711383 TI - [Karyosphere and extrachromosomal nuclear bodies in oocytes of the scorpionfly, Panorpa communis]. AB - Oocyte nuclear structures were studied for the scorpionfly Panorpa communis at different stages of oocyte growth, from pachytene to the first meiotic division. Using immunofluorescent and immunogold microscopy, we analyzed the nuclear distribution of RNA polymerase II, splicing factors and coilin. These factors were revealed in close association with perichromatin fibrils and, later, with some elements of the karyosphere and extrachromosomal nuclear bodies (NBs). Besides, it was shown that large amounts of P. communis oocyte NBs represent Cajal bodies (CBs) and contain CB marker protein, coilin, as well as RNA polymerase II, and in some cases an essential splicing factor, SC35. The presence of SC35 is commonly not characteristic of CBs in somatic cells. CB dynamics was traced in inactivated oocyte nuclei, during a gradual condensation of chromosomes and their final assembling into the karyosphere. It has been shown that coilin, RNA polymerase II and SC35 protein are common compounds shared by CBs and some granular material associated with these condensed chromosomes. CB remnants were demonstrated in the ooplasm after the breakdown of nuclear envelope before the first meiotic division. In inactivated oocyte nuclei, CBs serve presumably as storage compartments for some inactive components essential for gene expression. PMID- 16711384 TI - [Effect of alkaloid sanguinarine and a pharmaceutical preparation ukrain on modulation of vesicular membrane fusion and actin cytoskeleton of macrophages]. AB - A study was made of modulations of lysosome-phagosome fusion process and of fibrillar actin content in mouse peritoneal macrophages by an antitumor alkaloid sanguinarine and a derivative drug Ukrain. In addition, effects of these substances on in vitro polymerization of monomeric globular actin from rabbit muscle were investigated. Sanguinarine and Ukrain stimulated lysosome-phagosome fusion and increased the content of polymerized fibrillar form of actin in mouse macrophages. Effects of these substances were enhanced at their higher concentrations. Both sanguinarine and Ukrain induced in vitro polymerization of globular actin from rabbit muscle. A possible role of sanguinarine and Ukrain in changing vesicular membrane states during intracellular membrane interaction in lysosome-phagosome fusion process was discussed. The influence of these substances on actin polymerization and actin cytoskeleton rearrangement was evaluated. It could be supposed that sanguinarine and Ukrain may alter intracellular membrane transport. PMID- 16711385 TI - [A study of DNA depolyploidization and depolytenization of the heterochromatized gonosomal chromatin bodies in the secondary giant trophoblast cells of the field vole Microtus rossiaemeridionalis using cytophotometry]. AB - A study was made of the distribution of the heterochromatized gonosomal chromatin bodies (GCB) material in the course of nuclear fragmentation of secondary giant trophoblast cells resulting in polykaryocyte formation at the late stage of their differentiation. A simultaneous DNA cytophotometry in GCBs and nuclear fragments showed a progressive GCB DNA content decrease proportional to that of DNA content in nuclear fragments. DNA contents in the nuclear fragments corresponded to 2c, 4c and 8c. In most cases 1-2 GCBs were found in the nuclear fragments of different ploidy levels. Both the total DNA content in GCBs and the DNA content in separate GCBs well correlated with the ploidy levels of fragments. The data obtained demonstrate a regular, whole-genome distribution of chromosomal materials into the nuclear fragments exemplified by sex chromosome distribution in compliance with the ploidy of nuclear fragments. We discuss a possible mechanism of nuclear fragmentation that may ensure substantially a balanced genome of nuclear fragments without leading to mitotic cycle renewal in the giant trophoblast cell population. PMID- 16711386 TI - [Premature assembly of nucleolus-derived foci induced by a reversible hypotonic shock in metaphase CV1 and HeLa cells]. AB - The assembly of nucleolus-derived foci (NDF) in the cytoplasm of telophase cells is an early stage of nucleolus reassembly during mitosis. In current literature, significant attention is paid to the molecular composition of NDF and their participation in reassembly of the mature nucleolus. However, very little is known about mechanisms controlling the NDF formation. The authors have demonstrated for the first time that a reversible action of low ionic strength buffers (lypotonic shock treatment) on living mitotic human HeLa and green monkey CV1 cells triggers a premature assembly of NDF at metaphase. Like the true NDF, i. e., those assembled in telophase mitosis, NDF prematurally induced at metaphase contain RNA and proteins required for rRNA processing (fibrillarin, B23/nucliophosmin, C23/nucleolin), but lack UBF, an auxiliary factor of RNA polymerase I. We have assumed that a reversible action of hypotonic shock on metaphase cells may result in temporal increase in intracellular [Ca2+](i) that, in its turn, may induce a premature assembly of NDF under isotonic conditions. The structural integrity of the mitotic spindle apparently plays an essential role in the response of metaphase cells to hypotonic shock treatments. PMID- 16711387 TI - [Participation of granulosa cells in mediation of prolactin and somatotropin action on bovine oocyte-cumulus complexes in vitro]. AB - Maturation of bovine oocyte-cumulus complexes (OCC) in media derived following granulosa cell culturing with prolactin (PRL, 50 ng/ml) and somatotropin (ST, 10 ng/ml) was studied. A medium conditioned by granulosa cells in the presence of PRL or ST exerted a stimulating effect on the proliferative activity of cumulus cells. ST introduction into the granulosa cell culture also caused a decrease in the rate of cumulus cells with degenerated chromatin at a subsequent OCC culturing. At the same time, the expansion of cumulus did not depend on hormone availability in the culture medium for granulosa cells. When OCC matured in conditioned media, a short-term inhibition of oocyte meiosis reinitiation (after 6 h of culturing) was revealed in both the experimental groups, as compared to the control. Furthermore, the addition of ST and PRL to granulosa cell culture resulted in a subsequent decline in the rate of oocytes with signs of chromosome degeneration, observed as early as by 6 h of incubation and to be retained throughout the whole period of OCC culturing. In this case the earlier resumption of meiosis was associated with a higher rate of degeneration of the nuclear material in oocytes. The results of the present study suggest that granulosa cells may mediate, at least in part, PRL and ST impacts on in vitro maturation of bovine OCC, with no contact between OCC and granulosa cells being required for hormonal signaling. PMID- 16711388 TI - [Dynamics of microtubular cytoskeleton in higher plant meiosis. V. Late prometaphase. The general scheme of anastral spindle formation]. AB - By means of morphological analyses of meiotic abnormalities in pollen mother cells (PMCs) of cereal wide hybrids, haploids and meiotic mutants, the processes involved in cytoskeleton cycle at late prometaphase (a sub-stage of transition from chaotic figure to bipolar spindle) were studied. A significance of the four processes of late prometaphase--axial orientation, lateral association, consolidation and convergance of spindle fibers--is discussed. PMID- 16711389 TI - [Characteristics of p53 protein stabilization in cells of patients with ataxia telangiectasia after gamma-irradiation]. AB - Stabilization of P53 protein in cells isolated from patients with a grave hereditary disease ataxia-telangiectasia (AT), characterized by strongly enhanced sensitivity to ionizing radiation and impairment of cell cycle control after DNA damage, was studied. The level of expression of these reactions by patients may vary, and it tends to be linked with the severity of the disease. In all AT strains studied, both acquired by the authors and obtained from foreign colleagues, we observed the alteration of timing and character of stabilization of P53 protein, after the action of ionizing radiation in sublethal dosage, as compared to that in cells from healthy donor. PMID- 16711390 TI - [Antiapoptotic oncogene bcl-2 induces a program of senescence in E1A + c-Ha-ras transformants treated with adriamycin]. AB - Introduction of bcl-2 gene in EIA + c-Ha-ras-transformed rat embryo fibroblasts, which are unable to be arrested after damaging influences and possess high proapoptotic sensitivity, results not only in suppression of cell death but also in re-establishment of cell cycle block following DNA damage and serum starvation. Flow cytometry showed that E1A + c-Ha-ras + bcl-2-transformants treated with DNA-intercalator adriamycin are capable of being arrested at G1/S boundary for a long time (for less than 5 days). According to the growth curve data, the number of Bcl-2-overexpressing cells remanins constant for a week of cultivation with adriamycin. Clonogenic efficacy of E1A + c-Ha-ras + bcl-2-cells is brought to no already in 16 h after adriamycin addition. Apoptotic death, revealed by oligonucleosomic fragmentation of DNA, as well as cell death, occurring due to mitotic catastrophe, after adriamycin treatment are almost absent in Bcl-2-overexpressing transformants, as compared with parental E1A + c Ha-ras-transformants. Bcl-2 introduction in E1A + c-Ha-ras-transformants is accompanied by a rise of SA beta-Gal (Senescence Associated beta-Galactosidase) activity, which is commonly considered to be a marker of cell senescence. Adriamycin treatment of E1A + c-Ha-ras + bcl-2-transformants results in a much higher rise in SA beta-Gal activity, as compared with untreated cells. Co immunoprecipitation experiments demonstrated the introduction of Bcl-2 to result in formation of Bcl-2 complexes with early region E1A oncoproducts, which are thought to be responsible for proapoptotic susceptibility of E1A-expressing transformants. The data obtained lead to suggestion that bcl-2 transfer to E1A + c-Ha-ras-transformants may induce a switch from the cell death program on the program of senescence after DNA damage, due, presumably, to Bcl-2 interaction with the apoptosis activator the viral oncoprotein E1A. PMID- 16711391 TI - [The lack of cyclin-dependent phosphoprotein kinase Pho85p leads to defects in mitochondrial nucleoid transmission in yeast Saccharomyces cerevisiae]. AB - Cyclin-dependent phosphoprotein kinases (CDK) play essential role in the regulation of the progression through different phases of the cell cycle, being also involved in metabolic regulation and in actin cytoskeleton organization. The Saccharomyces cerevisiae CDK Pho85p is associated with 10 different cyclins and phosphorylates different substrates, among these transcription factors Pho4p and Gcn4p, and a protein involved in actin polymerization being a homologue of amphiphysin 1 of mammalian Rvs167p. It is known that mutations in the PHO85 gene have a multiple pleotropic effects, one of these effects being a rapid accumulation of mitochondrial [rho-] mutations observed on the background of gene PHO85 inactivation. In this study it was shown that the appearance of [rho-] clones is a result of some defects in mitochondrial nucleoid transmission from mother to bud cells. PMID- 16711392 TI - [The effect of immobilized fibronectin on karyotypic variability in the skin fibroblast cell subline of indian muntjac]. AB - The numerical and structural karyotypic variability has been investigated in the Indian muntjac skin fibroblast cell subline MT on cultivating cells on the fibronectin-coated surface. In cell subline MT, cultivated on the fibronectin coated surface for 1 and 2 days, the character of cell distribution for the chromosome number did not change. In 3, 4 and 8 days, the character of cell distribution for the chromosome number changed. These changes involve a significant decrease in frequency of cells with modal numbers of chromosomes, and an increase in frequency of cells with lower chromosome numbers. Many new additional structural variants of the karyotype (SVK) appear. The observed alterations seem to be due to both disturbances of mitotic apparatus and selection of SVK, which are more advantageous to changed culture conditions of the cell population. Detachment of cells from the fibronectin-coated surface, followed by a 1 day cultivation on a hydrophilic surface, commonly used for routine cell cultivation, does not restore the control cell distribution for the chromosome number, but cultivation in these conditions for 5 days restore control distribution. The frequency of chromosomal aberrations on cultivation on the fibronectin-coated surface for 3 and 4 days significantly increases, mainly at the expence of dicentrics (telomeric association). On prolongating the time of cultivation up to 8 days on the fibronectin-coated surface the frequency of chromosomal aberrations approaches the control value. Structural instability of chromosomes at cultivation on the fibronectin-coated surface demonstrates nonspecific reaction of "markerless" cell lines to unfavourable factors of the environment. We discuss possible reasons of differences in the character of karyotypic variability in cell lines of the Indian muntjac skin fibroblasts on cultivating on laminin and fibronectin. PMID- 16711393 TI - [Cytomorphological characteristics of Polypodium hydriforme and problems of myxozoan and cnidarian phylogeny]. AB - The present review analyses cytomorphological characters of the parasitic cnidarian Polypodium hydriforme, discriminating between those of bilateral (triploblastic) animals, common characters shared with the Myxozoa, and the unique characters of this species. Phylogenetic position of the group of parasitic cnidarians and of the class Polypodiozoa is discussed. A conclusion is made that the cytomorphological characters as well as 18S rDNA analysis of P. hydriforme and Myxozoa justify establishment of a new taxonomic group (a clade) of parasitic cnidarians (Endocnidozoa) uniting Polypodiozoa and Myxozoa (Zrzavy, Hypsa, 2003). The unique characters of P. hydriforme suggest that the phylum Cnidaria is more diverse than commonly supposed, and that P. hydriforme is not an aberrant cnidarian species but a relic organism, which might originally belong to the cnidarian class Polypodiozoa, which underwent reduction in the course of adaptation to parasitism. PMID- 16711394 TI - [Interactions in solution between cyclodextrins and statins]. AB - Cyclodextrins (CDs) are cyclic oligosaccharides, capable of forming inclusion complexes with hydrophobic molecules in aqueous solution and therefore, of improving the bioavailability of many drugs. They are biocompatible with living cells and do not cause toxic side effects. In this study, interactions in aqueous solutions between chemically different CDs and lovastatin and simvastatin, respectively, were investigated. It was found that the solubility of both statins can be considerably increased by the CD derivatives. The influence of the chemical structure of CD and of the temperature and pH on the solubility of the statin derivatives is described. Possible ways of increasing the solubility of these cholesterol-lowering drugs and their utilization in practice are also outlined. PMID- 16711395 TI - [Synthesis of thiophene and alpha-terthiophene derivatives with antiproliferative activity]. AB - We have synthesised a series of known alpha-terthiophene lead molecules with PKC (protein kinase C) inhibitory activity and the compounds were tested in cell proliferation assay on EGF-RTK (epidermal growth factor receptor protein tyrosine kinase) over-expressing tumour cell line (A431). We found that two of them had excellent antiproliferative activity. We prepared a focused molecule library around the thiophene and the terthiophene scaffold and examined these compounds in cell proliferation assay on A431. PMID- 16711396 TI - [Effect of tofisopam on CYP3A4 enzyme activity on human recombinant 3A4 supersome]. AB - Tofisopam is an anxiolytic agent of the BZD group, chemically 1(3-4 dimethoxyphenyl)-4methyl-5-ethyl-7,8 dimethoxy-5H-2,3-benzodiazepine. TZP differs from the traditional 1,4-benzodiazepines regarding the positions of the nitrogen atoms. Three clinical cases were reported where tofisopam increased the blood level of immunosuppressive agent leading clinically relevant adverse drug reaction and necessitating reduction of the dose of the drugs or discontinuation of the administration of tofisopam. The administered immunosuppressive agent is a substrate of the CYP3A4 system, so the effect of tofisopam on the CYP3A4 enzyme was investigated in vitro using human recombinant CYP3A4 supersome. Benzyoxy-4 (trifluoromethyl)-coumarin (BFC) was used as substrate. Tofisopam in 0.1, 0.25, 0.5, 0.75, 1 and 5 micromol/l concentrations inhibited dose dependently the enzyme activity. Activity inhibition rates were 4%, 29%, 40%, 56%, 61% and 94%, respectively and the IC50 was 0.8 micromol/l. The IC50 of positive control substance ketoconazole was 0.03 micromol/l. In in vitro experiments the inhibitory effect of tofisopam was lower than that of ketoconazole (potent CYP3A4 inhibitor) with an order of magnitude. According to the in vitro results it could be concluded that tofisopam is an inhibitor of CYP3A4 but to clarify the clinical importance of this inhibition further human clinical data are needed. PMID- 16711397 TI - [Dilemmas of the pharmacist, 7th edition of Hungarian National Formulary in practice]. AB - Most important event of year 2004 of the history of Hungarian pharmacy was the release of the 7th edition of the National Formulary. The general part of the new formulary expanded, remarks on the preparations are more detailed and dispensing technologies are more elaborated. Knowledge base on pharmaceutical substances and incompatibility is inserted as novelty. Following the principles of modern pharmacy practice the new National Formulary excludes pills, though it is still accepted as an alternative dosage form. Usage of tablets, hard gelatine capsules and medication stick as a new dosage forms are introduced. The aim of my study was to prepare and examine some new compositions of the new edition of the National Formulary to help the pharmacist's work. Results confirmed the novel solutions of the new National Formulary. PMID- 16711398 TI - [Investigation of the effects of drugs and UV radiation on biological and artificial membranes]. AB - Investigation of the effects exerted by different physical and chemical agents on biological and model-membranes is of prominent importance. Recently, a general trend can be observed in the formulation of drugs: incorporation of the drugs into liposomes. Knowledge of the molecular interactions between the transporting lipids and the incorporated agents is therefore very important. Nowadays, the increased environmental UV radiation requires investigation of the effects of the UV radiation exerted on biological membranes. Beside of modeling biological effects, studies on the effects of the UV radiation on model membranes can result in new knowledge on the stability of the liposomes containing phototoxic drugs. During my study, three different methods (EPR spectroscopy, DSC and light scattering measurements) have been applied to investigate the molecular interactions between drugs and the lipid molecules. Derivatives of the morphine as well as the (fluoro)quinolones mainly interact with the headgroups of the lipid molecules resulting in an increase of the molecular ordering of the lipids. My observation that drugs with protonable/deprotonable groups can modify the membrane-fluidity due to specific, local interactions with the lipid/stearic acid molecules of a membrane depending on the pH as well, call attention to choose optimal pH-interval for such drug formulations. Investigating the UVA effect on human fibroblast cell line I concluded that a decrease in the membrane fluidity due to UVA radiation can be detected only at doses higher, than 150 kJ/m2, and close to the lipid head groups. PMID- 16711399 TI - [Physical ageing of amorphous polymeric excipients I. physicochemical principles]. AB - Most of the polymeric excipients applied in pharmaceutical technology are amorphous, which, as a result of physical ageing, can lead to changes in the stability of these materials and dosage forms prepared from them. For the tracking of physical aging and understanding its consequences, a complex knowledge of the physicochemical properties and behaviour of amorphous polymers is necessary. In the case of these materials, three single-phase physical states can be distinguished: glassy, rubbery and viscous. The transition from glassy to rubbery state occurs at the glass transition temperature, the change of which as a function of the storage conditions provides information about the physical ageing of the material. The ageing process is usually accompanied by enthalpy and volume relaxation, which are considerably influenced by the presence of different plasticizers, e.g. water. These materials usually change the glass transition temperature of the polymer, which is a result of their effect on the free volume of the system. In view of the hygroscopic behaviour and water-uptake mechanisms of polymers, the probability and extent of structural changes caused by physical aging can be predicted. PMID- 16711400 TI - Problem areas and inconsistencies in the WHO classification of thymoma. AB - The morphologic classification of thymoma has undergone numerous revisions in recent years and has been under continuous debate for the past several decades. With the introduction of the World Health Organization (WHO) schema for the classification of thymic epithelial neoplasms in 1999, a major step was taken toward achieving uniformity in the nomenclature of these tumors. A more recent iteration of the WHO morphologic classification of thymic epithelial neoplasms has now been published. Although these efforts have certainly contributed to clarifying many issues related to the pathology of thymoma, several problem areas and inconsistencies still remain surrounding this proposed schema. The purpose of this review is to address these problem areas, in particular as it relates to terminology, histopathologic criteria for diagnosis, and the prognostic significance for the various categories of the WHO schema. PMID- 16711401 TI - Thymic carcinoma: update of current diagnostic criteria and histologic types. AB - Thymic carcinomas are rare tumors thought to derive from thymic epithelium. Because of the complex embryological origin of the thymus, whose development includes contributions from the third and, to a lesser extent, the fourth pharyngeal pouches, thymic carcinomas are endowed with great morphologic heterogeneity. A large number of histologic types have been described that resemble tumors arising in other organs. Unfortunately, no definitive pathognomonic histological features or immunohistochemical markers are associated with these tumors, making them a real challenge for diagnosis. Because of their close similarity with tumors arising at other organs, the diagnosis of thymic carcinoma must be regarded, for the most part, as a diagnosis of exclusion. This review will focus on current criteria for diagnosis of these tumors, with a review of the various histopathologic appearances that they can adopt. PMID- 16711402 TI - Cytopathology of thymic epithelial neoplasms. AB - A cytologic diagnosis of thymoma is extremely challenging. In part, this is because the tumor is uncommon and aspirates are infrequently encountered, a technically proficient interventional radiologist is needed, epithelial cells may be difficult to recognize in lymphoid rich aspirate smears, and there is inherent sampling error in a tumor that frequently displays heterogeneous histopathology. Critical to the cytologic diagnosis of most WHO Type B thymomas is the recognition of a distinct population of epithelial cells mixed with lymphocytes. This is more easily accomplished using Papanicolaou or H&E stains, and often requires a cytokeratin stain for verification (in the correct clinical-radiologic context) because these cells are cytologically bland and have a varying amount of cohesiveness. WHO Type A thymoma may contain only epithelial cells and thus mimic a spindle cell neoplasm, or mesothelial cell clusters. Limitations of the cytologic method include an unproven ability to definitively separate thymoma into specific WHO subtypes using cytology alone, and to determine capsular invasion. Non-neuroendocrine thymic carcinomas mimic their extra-thymic counterparts in cytologic aspirates, and their malignant nature is usually readily recognizable. Thymic neuroendocrine carcinomas (NEC) are also cytologically identical to their more common pulmonary sites of origin, but identification of moderately-differentiated NEC is generally not possible. PMID- 16711403 TI - Primary neuroendocrine carcinomas of the mediastinum: review of current criteria for histopathologic diagnosis and classification. AB - Primary neuroendocrine neoplasms occurring in the mediastinum are rare. The spectrum of such tumors can range from low- to high-grade neoplasms. The histogenesis of these tumors is varied, and some of them may originate from ectopic tissues in the mediastinum whereas others represent tumors native to the thymus. Primary thymic neuroendocrine carcinomas therefore need to be separated from other neuroendocrine neoplasms of the mediastinum, namely ectopic parathyroid tumors and paragangliomas. The histopathologic classification of primary neuroendocrine carcinomas of the mediastinum is still under debate and continues to be controversial, as is attested to by the different publications on the topic. Some authors continue to use old terms such as "carcinoid" and "atypical carcinoid" to designate these lesions, whereas others favor the use of a more unifying nomenclature that acknowledges the malignant nature of these lesions, namely that of neuroendocrine carcinoma. However, one aspect that all agree on is that, when they occur in the thymic region, these tumors should be considered as aggressive neoplasms capable of local recurrence and distant metastasis. This review will center on the morphologic spectrum of neuroendocrine carcinomas of the thymus, with special emphasis on diagnostic features and criteria for classification. PMID- 16711405 TI - Cystic lesions of the mediastinum. AB - Cysts of the mediastinum comprise a relatively diverse group of lesions that include neoplastic and nonneoplastic constituents, the latter of which are largely congenital in nature. Therefore, a grasp of their clinicopathologic characteristics is bolstered by knowledge of which tumors in the chest may undergo cystic change, as well as the embryologic development of the thoracic organs. That information, as well as radiological and pathologic features of these lesions, is discussed in this review. PMID- 16711404 TI - Mediastinal germ cell tumors. AB - Germ Cell Tumors (GCT) represent an important group of mediastinal tumors. Because of genetic characteristics and behavior have been divided in prepubertal and postpubertal tumors, and their origin remains controversial. The genetic changes are diverse, but the most frequent is i (12p), and can be associated to gonadal disgenesis. Histological classification is similiar to that of gonadal tumors and all histological types have been described in the mediastinum. Teratomas may undergo malignant transformation with the emergence of somatic tumors such as haematological malignancies, sarcomas, carcinomas and neurogenic tumors, and such transformation may occur in the primary site or in the metastases. Prognostic factors in mediastinal germ cell tumors include: age of the patient, histological type, distant metastases, stage, status of resection, level of serum tumor markers and proliferation markers. Immunohistochemistry is especially useful when the primary GCT is occult, to separate types of immature GCT, and to determine the lineage of malignant transformation which may be important to design treatment strategies. PMID- 16711406 TI - Bidding rule prompts trade groups' call to action. PMID- 16711407 TI - Soaring gas prices squeeze HHA, employee budgets. PMID- 16711408 TI - Continuous care probe hits hospices. PMID- 16711409 TI - Court rules VA medical center can not pay PAs more than NPs. PMID- 16711410 TI - Deplorable nursing care results in Medicare disqualification. PMID- 16711411 TI - OH: iodine used to treat wound despite allergy: summary judgment for defendant reversed. PMID- 16711412 TI - 'Whistleblower' fired by nursing home awarded damages. PMID- 16711413 TI - [Survey of study on the extraction, purification and determination methods of glycyrrhizic acid in Licorice]. AB - Extraction, purification and determination methods of glycyrrhizic acid in Licorice are surveyed in this paper. The extracting efficiency of dilute ethanol solvent for glycyrrhizic acid is higher than water extraction, and ammonia appended to ethanol solution can increase the effect of extraction. Ultrasound method, microwave-assisted method and supercritical CO2 fluid extraction are more effective than the conventional techniques, due to the short extraction time, low consumption of solution and energy. The modem analytical methods such as TLCs, HPLC and CE can determine the content of glycyrrhizic acid rapidly and exactly. PMID- 16711414 TI - [Advances in studies on active components and monomes of the traditional Chinese medicine targeted platelet]. AB - These active components and monomes inhibit thrombosis aimed directly at activation, adhesiveness and aggregation of platelet, thus preventing and curing ischemic cardiovascular and cerebrovascular diseases. Here we summarized the effect of active components and monomes of the traditional Chinese medicine targeted platelet on ischemic cardiovascular and cerebrovascular diseases, to provide references for drug investigation and clinical application. PMID- 16711415 TI - [Study on the stigma/pollen vigor and self-compatibility of Platycodon grandiflorum]. AB - OBJECTIVE: To provide the basal data for the breeding and cultivation of Platycodon grandiflorum. METHOD: The field investigation and pollination by bagging were carried out. TTC(2,3,5-triphenyl tetrazolium chloride) solution was used to test the pollen vigor. RESULT: The stigma life-span of P. grandiflorum was about 9 days, however the optimal time for pollination is 4-6 days after the petals opening, with the stigma was splitting lightly or significantly. When the petals opened, the anther began scattering pollen, and finished in the same day. The pollen vigor was about 81.4% at the beginning, and decreases to 27.6% three days later. The pollen vigor still remains 64.4% three days later, when the flower was kept in the desiccant. The natural fructification rate of self-flower pollination was 4.8%. The fructification rate and compatible index was about 62.7% and 54.6, respectively, when the self-plant-pollination performed by hand. They decreased to 12.8% and 6.5 when the pollination was implemented during the flowering period. CONCLUSION: Low fructification percentage of self-flower pollination attributes to the difference of maturing period of pistil and stamen, as well as the short pollen life-span of P. grandiflorum. The compatibility of self-plant-pollination is high during flowering period when pollination performed by hand. The life-span of the pollen can be prolonged significantly when keeping in the dry environment. PMID- 16711416 TI - [Study on tissue culture of Bolbostemma paniculatum]. AB - OBJECTIVE: To study the method of callus induction and culturing of Bolbostemma paniculatum and its relationship with accumulation of tubeimoside. METHOD: Stem and leaf were selected as explants. The effects of growth substances and their combinations at different concentrations on callus formation and propagation on MS and B5 basic medium were studied. RESULT: The optimal compositions of medium that induced calli from stems and leaves were the MS medium supplemented with 2,4 D 2.0 mg x L(-1), NAA 0.5 mg x L(-1) and BA 1.0 mg x L(-1) and the MS medium supplemented with 2,4-D 0.5 mg x L(-1) and NAA 2.0 mg x L(-1), respectively. The calli cultured on the MS medium supplemented with 2,4-D 2.0 mg x L(-1), NAA 0.5 mg x L(-1) and BA 1.0 mg x L(-1) were optimal for the accumulation of tubeimoside. A uniform and stable callus line could be formed from the calli cultured repeatedly on B5 medium supplemented with 2,4-D 2.0 mg x L(-1), NAA 0.5 mg x L(-1) and BA 1.0 mg x L(-1). CONCLUSION: Using stem and leaf as explants, the method of the callus induction and culture of B. paniculatum was established and it could provide some references for the production of tubeimoside by tissue and cell culture. PMID- 16711417 TI - [Effect of source-sink ratio changing on yield formation of Cynanchum bungei]. AB - OBJECTIVE: To study the effect of changing source-sink ratio on dry matter accumulation yield, distribution and interrelated physiological index of Cynanchum bungei. METHOD: Bud, fruit or side tress of C. bungei were picked artificially in the development stage. LAI, chlorophyll content, photosynthesis of single leaf, accumulation and distribution of dry matter at different and treatments were measured. The untreated plant was used as the control. RESULT: Although leaf decrease can be compensated by the increasing photosynthesis rate of single leaf, the LAI was small, chlorophyll decomposes quickly and these leaves showed the signs of early ageing. The matter accumulation was lower than that of control. Picking bud and fruit made photosynthesis rate of single leaf descend, chlorophyll decomposes slowly and its content keeps high, that inhibited ageing of the plant obviously. The LAI keeps larger, photosynthate was abundant. Dry matter accumulation and distribution to the root were higher than that of control and that of side tress picked. So, the yield with this treatment increased. CONCLUSION: Picking bud can increase LAI, postpone the decomposition of chlorophyll, and protract the functional leaves. Consequently, picking bud can add the accumulation of dry matter, increase proportion to radix and improve the yield and economy benefit. PMID- 16711418 TI - [Preparation of solid lipid nanoparticles loaded with Xionggui powder supercritical carbon dioxide fluid extraction and their evaluation in vitro release]. AB - OBJECTIVE: To study the preparation of solid lipid nanoparticles loaded with Xionggui powder-supercritical carbon dioxide fluid extraction and their evaluation in vitro release. METHOD: To prepare solid lipid nanoparticles (SLN) loaded with Xionggui powder-supercritical carbon dioxide fluid extraction (XG-CO2 SFE) using a hot dispersion- ultrasonic technique, establishing the best prescription of XG-CO2-SFE-SLN through orthogonal design methods using entrapment efficiency of nanoparticles as index, and investigating their physicochemical characterizations. The invro release action of SLN was studied in different dissolution mediums using dynamic dialyse method. RESULT: The best prescription was: phospholipid: F-68: stearie acid glyceride = 5: 2 : 1, the entrapment efficiency of nanoparticles was 96.3%, and the results revealed the nanoparticles were sphere like with the mean size of 245.8 nm, the mean Zeta potential was 33.5 mV. The in vitro release meet to Weibull distribution in physiological brine and to single-index model in pH 7.4 phosphate liquid (40% EtOH). CONCLUSION: The preparation method of the XG-CO2-SFE-SLN was appropriate, and the XG-CO2-SFE-SLN was released completely. PMID- 16711419 TI - [Optimization of formulation of Fufang Danshen immediate release tablet by colligation score]. AB - OBJECTIVE: To optimize the formulation of immediate release tablet. METHOD: The immediate release tablet was prepared by using dry granules. The preparation was optimized by using orthogonal design which took the flow property of granules, the hardness, the disintegrating time and the dissolution rate of the tablet as indices. RESULT: The optimized formulation contained 40% microcrystalline cellulose, 10% sodium carboxymethyl starch and 15% dextrin. The hardness disintegrating time and T50 of the tablet were 4.5 kg, 3 min, 5 min respectively. CONCLUSION: It is successful to prepare on immediate release tablet using the optimized formula above. PMID- 16711420 TI - [The content condition of effective composition in Sanhuang Xiexin decoction]. AB - OBJECTIVE: To investigate the content and existing conditions of baicalin, berberine and rhein in Sanhuang Xiexin decoction suspension. METHOD: Three active components (baicalin, berberine and rhein) in different prescriptions of original, absentone ingredient herb, and single herb were determined by HPLC. RESULT: There were baicalin 1,211 mg, berberine 276 mg and rhein 25 mg in 200 mL Sanhuang Xiexin decoction. Even though there will be precipitation if alkaloid is together with baicalin and some anthraquinones, no obvious interaction showed the affecting extract content between Scutellaria baicalensis and Coptis chinensis. The extracted rhein content can be increased obviously by Scutellaria baicalensis. Rheum patmatum and Coptis chinensis can suppress medicinal composition release of each other. CONCLUSION: The acid and basic components can affect each other and the acid components themselves also can affect each other. PMID- 16711421 TI - [Determination of 20 (S)-ginsengnoside Rh2 in the alkali-hydrolysis product of saponins from leaves of Panax qinquefolium by RP-HPLC]. AB - OBJECTIVE: To determine 20(S)-ginsengnoside Rh2 in the hydrolysis product of saponins from leaves of Panax qinquefolium. METHOD: The separation was performed on ZORBAX EXEND C18 column (4.6 mm x 250 mm, 5 microm), eluted with methanol and water (85:15) as mobile phase with the rate of 1.2 mL x min(-1) at 25 degrees C, the wavelength for measurement was 203 nm. RESULT: The calibration curve was linear in the range of 0.5-25 microg for 20(S)-ginsengnoside Rh2(r = 0.9999, n = 7). The average recovery was 99.7% (RSD= 1.0%). CONCLUSION: This method is simple, accurate, reliable and reproducible. The result shows that the transform ratio of 20(S)-ginsengnoside Rh2 is high by this hydrolysis method. PMID- 16711422 TI - [Determination of curcumol in Rhizoma Curcumae by GC]. AB - OBJECTIVE: To establish a method to determine the content of Curcumol in Rhizoma Curcumae. METHOD: The samples were determined by GC on a HP-5 column (0.32 mm x 30 m, 0.25 micron), Inlet temperature 200 degrees C, FID 250 degrees C, flow 1.0 mL x min(-1), splitless. Temperature programming started at 60 degrees C, holding for 4 min, then increased to 210 degrees C at a rate of 3 degrees C x min(-1). RESULT: The calibration curve of curcumol is linear over the range of 40.0-2,000 microg x mL(-1), r = 0.9997. The high and low additive recovery were 95.01% (RSD 2.52%), 96.46% (RSD 2.86%). CONCLUSION: This method was accurate and reliable with a good reproducibility, and the procedure of samples pretreatment is simple. PMID- 16711423 TI - [Studies on flavonoids of Andrographis paniculata]. AB - OBJECTIVE: To study the flavonoids of the aerial parts of Andrographis paniculata. METHOD: Twelve flavonoids from the 85% EtOH extract were isolated by the silica gel column, Sephadex LH-20 column, ODS column chromatography and HPLC, and their structures were identified by the spectal analyses and chemical evidences. RESULT: The 12 compounds were obtained and identified as 7-O methylwogonin (1), 7-O-methyldihydrowogonin (2), 5-hydroxy-7, 8, 2', 5' tetramethoxy-flavone (3), skullcapflavone-2'-methoxylether (4), 5-hydroxy-7, 8, 2', 3'-tetramethoxyflavone (5), 5, 4'-dihydroxy-7, 8, 2', 3'-tetramethoxyflavone (6), dihydroskullcapflavone (7), 5, 7, 8-trimethoxydihydroflavone (8), 5, 2' dihydroxy-7, 8-dimethoxyflvone (9), andrographidine C (10), 5, 7, 4' trihydroxyflavone (11), 5, 7, 3', 4'-tetrahydroxyflavone (12). CONCLUSION: Compound 6 is a new naturally occurring product isolated from A. paniculata, and compound 8, 11 and 12 are obtained for the first time from the Andrographis species. PMID- 16711424 TI - [Studies on the flavones in of Chrozophora sabulosa]. AB - OBJECTIVE: To study the flavone constituents in Chrozophora sabulosa (Xinjiang origin). METHOD: The compounds were extracted with 95% ethyl alcohol, isolated by various column chromatography and identified by spectroscopic methods. RESULT: Seven flavanoids were isolated and identified as quercetin (I), kaempferol (II), apigenin (III), chrysoerid (IV), isoquercitrin (V), chrysoerin-7-O-beta-D glucoside (VI) and quercetin-3-O-alpha-D-arabinfuranoside (VII). CONCLUSION: All of these seven compounds were obtained from this genus for the first time. PMID- 16711425 TI - [Studies on antiviral constituents in stems and leaves of Pithecellibium clypearia]. AB - OBJECTIVE: To study the antiviral constituents in the stems and leaves of Pithecellibium clypearia. METHOD: The constituents of P. clypearia were systematically separated with various chromatographic techniques in combination with antiviral activity monitoring. Their structures were elucidated by physical and chemical properties and spectral data. RESULT: Six compounds were isolated from P. clypearia and were identified as: tricetiflavan (5, 7, 3', 4', 5' pentahydroxylflavan) (1), myricitrin (myricetin-3-O-alpha-L-rhamnopyranoside) (2), quercitrin (quercetin-3-O-alpha-L-rhamnopyranoside) (3), quereetin (4), methyl gallate (5) and gallic acid (6). CONCLUSION: Compound 1 approximately 5 were obtained from this plant for the first time. Compound 4 was found to show an obvious anti-respiratory syncytial virus (RSV) activity. PMID- 16711426 TI - [Studies on involatile constituents of Mentha haplocalyx]. AB - OBJECTIVE: To promote its comprehensive utilization, the involatile constituents of Mentha haplocalyx were studied systematically. METHOD: The chemical components were isolated and purified by silca gel column chromatography and recrystallization. The chemical structures were elucidated on the basis of physico-chemical properties and spectral data. RESULT: Eight compounds were isolated and identified as: acacetin (I), tilianine (II), linarin (III), n-butyl beta-D-fructopyranoside (IV), ursolic acid (V), oleanolic acid (VI), beta sitosterol (VII), daucosterol (VIII). CONCLUSION: Compounds I approximately V were obtained from this plant for the first time. PMID- 16711427 TI - [Effects of Rehmannia glutinosa oligosaccharides on proliferation of 3T3-L1 adipocytes and insulin resistance]. AB - OBJECTIVE: To investigate the influence of Rehmannia glutinosa oligosaccharides (ROS) on the proliferation of 3T3-L1 adipocytes and insulin resistance. METHOD: 3T3-L1 preadipocytes were cultured, the proliferation of 3T3-L1 preadipocytes was detected by MTT method. Insulin resistant 3T3-L1 adipocytes cell model was induced by dexamethasone and the change of glucose concentration in cell culture was determined after ROS treatment. RESULT: In the high glucose DMEM culture media, MTT method showed that the absorbance at 570nm of 3T3-L1 preadipocytes was increased and that of 3T3-L1 adipocytes was decreased. ROS significantly increased glucose consumption in 3T3-L1 preadipocytes and adipocytes culture in a concentration-dependent manner. ROS improved the sensitivity of 3T3-L1 adipocytes to insulin. CONCLUSION: ROS can promote the proliferation of 3T3-L1 preadipocytes, inhibite the proliferation of 3T3-L1 adipocytes, and also, significantly improve insulin resistance induced by dexamethasone. PMID- 16711428 TI - [Effects of Zhikeping on aortic oxyradical and the expression of ICAM-1 in a rat model of experimental early atherosclerosis]. AB - OBJECTIVE: To investigate the effects of Zhikepingon the oxyradical andintercellular adhesion molecule-1 (ICAM-1) of experimental early atherosclerosis. METHOD: The model of SD rat early atherosclerosis was induced by cholesterol diet. The suspension of Zhikeping and simvastatin were administered intragastrically, respectively. After 10 weeks, the serum lipids, superoxide dismutase (SOD) and maleic dialdehyde (MDA) were detected by automatic biochemistry analyzer. ICAM-1 and its expression of mRNA in aortic wall were detected- by immunohistochemistry and RT-PCR, respectively. Aortic histomorphology was cbserved by HE stainning. RESULT: The results showed that the serum lipids, superoxide dismutase (SOD) and maleic dialdehyde (MDA) of treated groups were obviously improved as compared with those of the control group. The tissue pathological damage was improved indifferent degree, and ICAM-1 and its expression of mRNA was decreased obviously. CONCLUSION: It is suggested the mechanism of anti-atherosclerosis of Zhikeping have close relationship with the function of its anti-oxidizing and anti-adhesiveness that can protect aortic endothelial cell. PMID- 16711430 TI - [Effects of Yifuning capsule on blood lipids of ovariectomized hyperlipidemia rats]. AB - OBJECTIVE: To observe the effects of Yifuning (YEN) capsule on blood lipids of ovariectomized hyperlipidemia rats. METHOD: Fifty-six female mature Sprague Dawley rats were randomized into 7 groups: normal control group, model control group, diethylstilbestrol tablets (DT) group, Xuezhikang group, YFN high, middle and low dose groups. The ovariectomized rats were fed on high fat diet and administrated with the drugs for 3 weeks, then were killed and estimated body weight, liver index and five items of blood lipid (TC, TG, HDL-C, LDL-C, VLDL) by test kit. Enzyme (such as HP, LDL, and whole lipase) was detected too. RESULT: The weight and liver index of model control group increased obviously as compared with normal group. YFN could reduce TG, TC, and LDL-C (P < 0.05) of ovariectomized hyperlipidemia rats obviously and increase HP, LDL and whole lipase (P < 0.05) on the other hand. CONCLUSION: YFN can ameliorate blood lipids of ovariectomized hyperlipidemia rats. PMID- 16711429 TI - [Study on the function of decoction for invigorating the kidney and improving blood circulation to thrombosis on rabbit blood stasis model]. AB - OBJECTIVE: To evaluate the effect of decoction for invigorating the kidney and improving blood circulation to thrombosis and pathology on rabbit blood stasis model. METHOD: Thirty rabbits were ramdomly divided into normal group, model group, high dose group, low dose group and Xue Shuan Ning group. Tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor (PAI), fibrinogen (Fbg) and D-dimer (DD) were investigated after those rabbits had been treated. One rot was solected randomly from each group to observe pathological changes. RESULT: There were significant differences in t-PA, PAI, Fbg and DD between normal group and other groups is very obvious (P < 0.01) . Between groups of high dose low dose Xue Shuan Ning and model, the statistical differeces were significant, as well as between groups of high dose, low dose and Xue Shuan Ning groups (P < 0.05). However, there was no statistical difference between high dose group and high dose group (P > 0.05). The pathological changes in model group were most serious, those in Xue Shuan Ning were less serious. There were slight pathological changes in high dose group and low dose group. CONCLUSION: Models ware made successfully. High dose group and low dose group have stronger effect on thrombosis than Xue Shuan Ning group. PMID- 16711431 TI - [Experimental study on the Sanxiansinli capsule against fatigue and viability of mice]. AB - OBJECTIVE: To investigate the effect of the Sanxianxinli capsule against fatigue and viability of mice. METHOD: The mouse shinning, burden swimming, heat resistant, cold resistant and tolerating anoxia were detected by experiment. RESULT: The results showed that the Sanxansinli capsule have function of improveing the stamina, cold resistant, heat-resistant and tolerating anoxia,and prolonging were the survival time in cold, high temperature and anoxia. CONCLUSION: The Sanxianxinli capsule have function against fatigue and can also improve the resistibility of body. PMID- 16711432 TI - [Effects and mechanism of total saponins of Psammosilene tunicoids against rheumatoid arthritis]. AB - OBJECTIVE: To observe the antiarthritic effects and the possible mechanism of total saponins of Psammruosilene tunicoids (TSPT) against rheumatoid arthritis (RA). METHOD: After establishing AA rat model, the TSPT'S antiarthritic effects and mechanism against RA were studied through observing the changes of ankle swelling, arthritis index and levels of IL-1beta and TNF-alpha after medication. RESULT: TSPT could effectively inhibits articular swelling, decrease arthritis index and regulate down the content of IL-1beta and TNF-alpha in the inflammatory tissue soak of AA rats. CONCLUSION: TSPT has good antiarthritic effects and the possible mechanism may be related to its down-regulation of IL-1beta and TNF alpha. PMID- 16711433 TI - [Protection of extract from Cuscuta japonica on human sperm acrosome and ultrastructure]. AB - OBJECTIVE: To study the oxidation damage of active oxygen (ROS) to human sperm acrosome and ultrastructure, and study the function mechanism about Cuscuta japonica treating male's infertility and asthenoospermia. METHOD: By using the Percoll gradient centrifugation, the sperm with normal physiological function were selected for the normal sperm model, and the sperm suspension were divided into the normal group, the model group, the positive control group (Vitamin C group), and the lugh, the median and the low dose gvoups of C. japonica. The ROS made from hypoxanthine-xanzine xanzine(HX-XO) and different content (0.125, 0.25, 0.5 g x mL(-1)) of extract were incubated with sperm in the oxygen environment. The acrosomic integrity rate were calculated and the sperm acrosome and ultrastructure were observed. RESULT: The content (0.125, 0.5 g x mL(-1)) of extract had no obvious difference as compared with Vitamin C (0.25 mg x mL(-1)) in protecting the acrosome and ultrastructure, but the content (0.25 mg x mL(-1)) of extract was significantly better than Vit C (P < 0.001). CONCLUSION: The suitable content of extract from C. japonica can significantly protect the sperm membrane, the acosomic structure and the mitochondrion function from the damage caused by ROS. PMID- 16711435 TI - [Genotypic analysis of familial dilated vestibular aqueduct syndrome]. AB - OBJECTIVE: To analyze clinical manifestation of patients from two families with dilated vestibular aqueduct syndrome (DVAS). Their genotypic patterns were discriminated with the genetic testing methods for PDS gene. METHOD: The twin sisters from pedigree and the brother and sister from pedigree 2 all suffered from sensorineural hearing loss. All patients from these two families were confirmed to have enlarged vestibular aqueduct by temporal bone CT scan. Exon 7+8 and their flanking area sequences were analyzed by direct sequencing. All other exons and their flanking sequences were screened by DHPLC. The amplicons of exons which showed abnormal DHPLC wave pattern were further sequenced. RESULT: The parents of two families are non-consanguineous, hearing normal couples with no other family members with hearing loss. Both families have two offspring with DVAS. The clinical features of all patients were progressive hearing loss, good communication ability but with thickness pronunciation. Both twin sisters from pedigree are homozygous for the splice site mutation (IVS 7-2 A-G) affecting the 3 splice site consensus sequence of intron 7. The brother and sister of pedigree 2 are compound heterozygotes for 1199 ins T mutation and a missense mutation (1229 C-T). CONCLUSION: Familial DVLS is typical hereditary disease. The patients with DVLS have similar clinical manifestation and the same genotypic changes in PDS. PMID- 16711434 TI - [Experimental study on effects of Shenshuai compound medicine on acute renal failure rats and secretion cell factors]. AB - OBJECTIVE: To observe the effect of the experiential compound prescription Shenshuai compound medicine (SSCM) on acute renal failure (ARF) rats, and expose its mechanism. METHOD: Male SD rats were allocated into four groups at random. Except normal group, the others were injected glycerin into the musculi to induce a model of ARF. At the same time, rats in normal and model groups were given a dose (10 mL x kg(-1)) of normal saline; rats in the other two groups were given verapamil (Vp) 40 mg x kg(-1) x d(-1) and SSCM 22.5 mg x kg(-1) x d(-1) respectively by gastric gavages. In this way they were killed at 24 h after injecting glycerin. RESULT: In contrast with model group, SSCM and Vp could ameliorate the renal function of acute tubular necrosis (ATN) rats markedly, in a way protect the renal inherent cell ultrastructure such as tubular epithelial cell etc at ATN early-stage; especially SSCM could enhance NO contents in serum, and was reduce ET levels inplasma, evidently cut down TNF-alpha contents in serum, and was partly superior to Vp. CONCLUSION: It is indicated that SSCM can adjust thebalance of contract and stretch blood vessal active substance and clear away initiate inflammatory medium, consequently alleviate renal pathological changes, prevent and treat ARF. PMID- 16711436 TI - [Multi-channel cochlear implantation in patients with congenital inner ear malformation accompanied middle ear malformation]. AB - OBJECTIVE: To report the multi-channel cochlear implantation in three patients with congenital inner ear malformation accompanied middle ear malformation, including CI surgery and postoperative outcomes of hearing and speech recovery. METHOD: Multi-channel cochlear implantation surgeries have been performed in three cases with congenital inner ear malformation accompanied middle ear malformation in our hospital from May 1995 to May 2002. The transmastoid approach was performed to the cases with congenital inner ear malformation accompanied middle ear malformation. The postoperative sound field hearing tests were taken to the cases three months after the cochlear implantation. RESULT: Multi-channel cochlear implantation was successfully performed in three cases with congenital inner ear malformation accompanied middle ear malformation. The 27, 28, 32 electrodes were inserted respectively in three cases. The intraoperative "Ozze" occurred in three patients. No severe intraoperative and postoperative complications occurred in three cases. All postoperative hearing thresholds were at 35-40 dBHL. CONCLUSION Multi-channel cochlear implantation could be performed in the cases with congenital inner ear malformation accompanied middle ear malformation through the transmastoid approach. The postoperative outcomes were satisfied. PMID- 16711437 TI - [Comparison of Epstein-Barr virus in nasopharynx tissue and desquamatine cell for diagnosis of nasopharyngeal carcinoma]. AB - OBJECTIVE: To develop a method for rapid screening and primary diagnosis of the nasopharyngeal carcinoma (NPC). METHOD: We detected the Epstein Barr virus (EBV) DNA from nasopharynx tissue and desquamative cells in 144 suspicious NPC patients using polymerase chain reaction (PCR). The results were compared with the pathologic diagnosis. RESULT: The sensitivities were 92.16% and 86.27% respectively, the specificities were 80.95% and 71.43% respectively. There was no significant difference between the results determined by the EBV DNA from nasopharynx tissue and desquamative cells using PCR. CONCLUSION: The detection of EBV DNA from nasopharynx tissue and desquamative cells of the NPC by PCR is a simple, safe, rapid, specific and sensitive technique. It is useful in screening, primary diagnosis, evaluation of the curative effect of the NPC. PMID- 16711438 TI - [Analysis of air caloric testing results for patients suffering from tympanic membrane perforation with vertigo]. AB - OBJECTIVE: To analyze characteristic of air caloric testing for patients with tympanic membrane perforation and to assess the availability of this testing for the patients. METHOD: Forty-three cases of chronic otitis media with tympanic membrane perforation and vertigo were investigated, in which there were 37 cases with single ear perforation and 6 cases with bilateral perforation. The patients were performed by air caloric testing, to observe the characteristic of nystagmus,slow phase velocity (SPV) and unilateral weakness (UW). RESULT: Four types of nystagmus, including enhancing, "normal", weakening and inversion, were observed with bilateral caloric air stimulation in ears with tympanic membrane perforation. Nystagmus of perforated ear presented SPV increased exceeding normal range or non-perforated ear and UW > 15% (15 cases, 17 ears), which was called enhancing type; normal nystagmus SPV in bilateral ear and UW < 15% (7 cases) was called "normal" type; weakened nystagmus SPV in perforated ear to non-perforated ear and UW > 15% (9 cases) was called weakening type; normal nystagmus in non perforated ear and inverted caloric nystagmus in perforated ear (12 cases, 14 ears) was called inversion type. In patients with chronic otitis media recurring lately, inverted nystagmus was observed in perforated ears with effusion and humidity in the tympanic cavity, the other types were observed in patients in stillness stage of otitis media. CONCLUSION: Bilateral caloric air became strong stimulation for the perforated ear, nystagmus of perforated ear with normal function of semicircular canal was stronger than that of the non-perforated ear. While the perforated ear with undermined function of semicircular canal, nystagmus could present normal or weakened. In the patients with otitis media in active stage, hot air stimulation could be turned to cool stimulation response so as to cause inverted nystagmus. Air caloric testing can be used to assess vestibular function of the patients with tympanic membrane perforation and vertigo. PMID- 16711439 TI - [Six cases of adenoid cystic carcinoma in external auditory canal]. AB - OBJECTIVE: By investigating the clinical and pathological features of adenoid cystic carcinoma in external auditory canal, we can improve our ability of diagnosis and treatment. METHOD: A retrospective analysis was performed on 6 patients with adenoid cystic carcinoma in external auditory canal, including clinical manifestations, the therapy as well as the follow-up results. All cases underwent subtotal temporal bone resection or other aggressive surgical excision. RESULT: All cases were alive for a period from 8 to 21 months after subtotal temporal bone resection or other aggressive surgical excision, without any signs of regional lymph node metastasis and distal metastasis. CONCLUSION: Adenoid cystic carcinoma in external auditory canal grows slowly, so patients can live with the tumor for a long time. But the long-term prognosis is poor. It is necessary to manipulate aggressive surgical excision at the first time to improve cure rate. PMID- 16711440 TI - [Analysis of auditory function on normal growth children in city and rural areas]. AB - OBJECTIVE: To study the auditory function of Chinese normal growth children, and to get a first hand data for protecting children's hearing. METHOD: The normal growth children (6 to 12 years old) in Qinling mountain area, pasturing area around Qinghai Lake and Nanjing city were included in the study, and their whole frequency auditory thresholds were examined on the spot. The auditory thresholds were analyzed. RESULT: There was significant difference between the boys' and girls' auditory thresholds among nearly the whole frequency in Qinling mountain. And there was significant difference between the boys' and girls' auditory thresholds among high frequency and super high frequency in pasturing area around Qinghai Lake, while the girls' auditory function was better than the boys'. In contrast, in Nanjing city no difference was found between the boys' and girls' auditory thresholds. The auditory thresholds in high frequency and super high frequency of the boys in Nanjing city were lower than those of the boys in mountain area and plateau pasturing area, while there was no difference between the girls except for low frequency. CONCLUSION: Worse auditory function was found in Chinese rural areas, especially among the boys. According to the local residents' living condition, the children in remote area nearly had no chance to receive the modern industrial noise and other strong noise. So the firecrackers' noise perhaps made this situation. And the education to avoid the firecrackers' noise must be put into practice in Chinese rural area since childhood. PMID- 16711441 TI - [The constrastive analysis on distant metastasis before and after radiotherapy of nasopharyngeal carcinoma]. AB - OBJECTIVE: To explore the duration related to distant metastases of nasopharyngeal carcinoma (NPC). METHOD: One hundred and three cases with confirmative diagnosis of NPC, in which the distant metastasis had happened before radiotherapy, were called the natural metastasis group. Another one hundred and thirty-eight cases, in which the distant metastasis had happened after radiotherapy, were called the metastasis after radiotherapy group. The clinical data and appearance time on distant metastasis from the two groups were analyzed. RESULT: It took 10.78 months (95% CI: 8.68-12.88) on average in the natural metastasis group from the primary symptom to the distant metastasis, and 20.77 months on average in the metastasis after radiotherapy group. Comparing the two groups, there was highly remarkable difference (P < 0.01). The distant metastasis happened in 91.26% of the cases in the natural metastasis group in 2 years. And in 1 year and in 2 years, the distal metastasis incidences of the natural metastasis group was higher than those of the metastasis after radiotherapy group by 34.41% and 28.94%, respectively. The distant metastasis happened in all cases in the natural metastasis group in 5 years, while in 2.9% of the cases in the metastasis after radiotherapy group beyond 5 years. There wasn't any obvious difference in the patients' sex and the appearance rate of the primary symptom (P > 0.05). But in the metastasis after radiotherapy group, the patients were younger and the T and N stages were earlier. CONCLUSION: The distal metastasis in the metastasis after radiotherapy group was at least 1-2 years later than that in the natural metastasis group. It was perhaps relative to the younger patients and the earlier stages. PMID- 16711442 TI - [Assessment of adenoid size in children by flexible fibrescope examination]. AB - OBJECTIVE: To assess the adenoid size in children by flexible fibrescope examination and outline an effective therapeutic program according to the examination. METHOD: In this study, the flexible fibrescope examination of the nasal cavity and nasopharynx and a lateral skull radiograph examination were performed in 130 children with snoring and the same examinations were performed in another 30 children without snoring. RESULT: The size of adenoid was negatively associated with the degree of snoring. (P < 0.01). There is a high correlation between the flexible fibrescope examination and the lateral skull radiograph examination (r = 0.435, P < 0.01). CONCLUSION: The application of the flexible fibrescope examination of the nasal cavity and nasopharynx has proved to be of great clinical value, especially in the selection of children for adenoidectomy. It is an examination with minor invasion which can be well tolerated by most children, giving more information than a lateral skull radiograph and avoiding unnecessary radiation. PMID- 16711443 TI - [Inspecting the cochlear scala tympanic with flexible and semi-flexible micro endoscope]. AB - OBJECTIVE: Flexible and semi-flexible micro-endoscopes were used in cochlear scala tympani inspection , to explore their application in inner ear examination. METHOD: Fifteen profound hearing loss patients preparing for cochlear implant were included in this study. During the operation, micro-endoscopy was performed after opening the cochlear scala tympani. And 1 mm diameter semi-flexible micro endoscope could go as deep as 9 mm into the cochlear scala tympani, while 0. 5 mm diameter flexible micro-endoscope could go as deep as 25 mm. The inspecting results were compared with video recording. RESULT: Using 0.5 mm flexible micro endoscope, we canould check cochlear scala tympani with depth range of 15-25 mm, but the video imaging was not clear enough to examine the microstructure in the cochlear. With 1 mm diameter semi-flexible micro-endoscope, we could reach 9 mm deep into the cochlear. During the examination, we found 3 cases with calcification deposit in osseous spiral lamina, l case with granulation tissue in the lateral wall of scala tympani, no abnormal findings in the other 11 cases. CONCLUSION: Inspecting the cochlear scala tympani with 0.5 mm flexible micro endoscope, even though we can reach the second circuit of the cochlear, it is difficult to find the pathology in the cochlear because of the poor video imaging. With 1 mm semi-flexible micro-endoscope, we can identify the microstructure of the cochlear clearly and find the pathologic changes, but the inserting depth was limited to 9 mm with limitation to examine the whole cochlear. PMID- 16711444 TI - [Study on effect of lead exposure to blood lead level and hearing in young offspring through gestation-rat]. AB - OBJECTIVE: To study the effect of lead exposure to the blood lead level and the hearing in young offspring through gestation-rat. METHOD: Experiments were carried out on 42 Wistar rats, half female and half male, randomly divided into five couple groups (A, B, C, D, E). The group A, as a normal control group, was fed with a standard diet. The group B, C, D and E were fed with a standard diet, additionally with 0.03%, 0.090%, 0.27% and 0.81% lead acetate added respectively, and then fed with standard diet after birth. DPOAE, ABR and blood lead measurement were conducted on the parents rats and the neonatal offspring rats at first week and at the third week respectively. RESULT: The blood lead level of neonatal rat reported positively related with that of the parent rats (r = 0. 5817). DPOAE test result: Both parent rats and neonatal rats presented a normal cochlea feedback curve. ABR test result: The period of the group D showed a significant difference from that of the normal control group A. Reaction threshold (dBpeSPL) of the neonatal rat group cl (at the first week) and group c3 (at the third week) showed a notable difference from that of the normal control group al and a3 (c >a). CONCLUSION: The blood lead level of a fetus rat is related with the exposure degree to lead during the pregnant period of mother rat. Low level lead exposure inside uterus during pregnant period will result easily in damage on the hearing nerve center segment and hearing conduct of a fetus rat for they are very sensitive. PMID- 16711445 TI - [Comparison study of hair cell markers in chicken inner ear]. AB - OBJECTIVE: To supply valuable markers for hair cell development and regeneration. METHOD: Cryosections of new hatched chicken inner ear were made. The position and intensity of Hair Cell Antigen, Espin, and Myosin7a expression were detected by indirect immunocytochemistry. RESULT: Hair Cell Antigen, Espin, and Myosin7a were all expressed in both auditory and vestibular hair cell. Hair Cell Antigen was located in the apical part of hair cell and hair cell bundles. Espin was expressed in the hair cell bundles and cuticular plate. Myosin7a immu no-staining resulted in the typical cytoplasm staining. CONCLUSION: Hair Cell Antigen, Espin, and Myosin7a are ideal markers for hair cell and can be used for studying the hair cell development and regeneration. These markers combined with F-actin staining could give a better result for qualitative and quantitative investigation for hair cell development and regeneration. PMID- 16711446 TI - [Experimental study of primary culture methods for spiral ganglion cells of newborn rats]. AB - OBJECTIVE: To optimize culture condition of spiral ganglion cells (SGCs) in vitro and to obtain highly purified SGCs. METHOD: The spiral ganglions from newborn rats were digested in 0.25% trypsin and 0.001% DNase. The SGCs suspension was plated at a density of 10(9) cells/L. The cells were kept in serum free medium (DMEM/F12+B27 supplement). And 5 micromol/L cytosine arabinoside (Ara-C) was added to the medium on the second day and maintained for 48 hours. Serum medium with or without Ara-C was set as a control. The morphology and the purity of SGCs were observed under microscope. RESULT: Highly purified SGCs can be harvested in serum free medium (DMEM/F12+B27 supplement) with Ara-C. The SGCs were identified with mouse anti neurofilament protein antibody by immunohistochemistry methods. CONCLUSION: The method used in this study is an optimal means to culture and purify SGCs that can meet the needs of further study. PMID- 16711448 TI - [Dermatology, a medical/surgical specialty to respond to the increasing incidence of skin tumors]. PMID- 16711447 TI - [An in vitro investigation of the regulation mechanism to the blood-labyrinth barrier permeability of guinea pig inner ear]. AB - OBJECTIVE: To investigate the regulation mechanism of the blood-labyrinth barrier permeability of guinea pig inner ear. METHOD: The cultured endothelial cell was derived from the stria vascularis from guinea pig inner ear. We exposed guinea pig inner ear capillary endothelial cell monolayers to substances known to increase or decrease cyclic AMP and measured their effect on endothelial permeability to albumin and Evan's blue and endothelial cell cyclic AMP and F actin concentrations. RESULT: Cholera toxin (CT) led to a remarkable increase in cyclic AMP concentration and a marked reduction in F-actin concentration which was associated with remarkable reduction in albumin and Evan's blue transfer across endothelial monolayers; In contrast, katlex led to an obvious reduction in cyclic AMP concentration and a marked increase in F-actin concentration which was associated with obvious increase in albumin and Evan's blue transfer across endothelial monolayers. CONCLUSION: The results indicated that cyclic AMP is an important determinant of the blood-labyrinth barrier in vitro. The barrier function changes are probably associated with apparent endothelial shape changes and qualitative and quantitative changes in F-actin. PMID- 16711449 TI - [Mohs micrographic surgery: concept, technic and indications]. AB - Mohs Micrographic Surgery is a technic to treat severe cutaneous carcinoma. By performing a microscopic and topographic analysis of the whole tissue excised, both a reduction of the surgical margin and margins in normal tissue is obtained. The major indications of micrographic surgery are recurrent basalcell carcinoma after surgery or radiotherapy, not fully excised or localized on the nose and the mediofacial region, some spinouscell carcinoma and dermatofibrosarcoma protuberans. PMID- 16711451 TI - [Surgery for ingrowing toenails]. AB - Ingrown toenail is a relatively frequent condition of the great toe affecting usually young adults. Surgery is most often required but the rate of recurrences is high with the risk of definitive functional and esthetical consequences. Among the numerous surgical procedures described, none is unanimously considered as the most effective. As for all dermatological surgery, the procedure must be not only curative but also esthetically acceptable. The surgeon's skills and experience are determining factors. A review of the most common procedures is presented. PMID- 16711450 TI - [The role of surgery in the treatment of acne scars]. AB - Severe acne lead to inesthetic scars on the face of young adults. A new classification based on the morphology of acne scars allows to establish the more appropriate therapeutic strategy. The surgical technics are: punch-excision, punch-elevation, subcission, dermal graft or adipose tissue autotransplantation. Skin resurfacing by dermabrasion, CO2 or Erbium/YAG laser or laser remodeling could be performed as a complement of the surgery. By associating these treatments acne scars are greatly improved. PMID- 16711453 TI - [Pruritus--causes, diagnostics and treatment]. AB - Diagnosis and treatment of pruritus is still a great problem. The pathophysiologic principles are mostly unknown, however the progress in neurosciences also added several new neurophysiologic explanations for pruritus. There exist several different types of pruritus and also the treatment of the different forms varies. The treatment of chronic pruritus is not the same as the treatment of acute pruritus. Usually more than one pathological disorder leads to severe pruritus and therefore the diagnostic of the different forms of pruritus and the treatment needs an interdisciplinary approach. One of the major causes of itch is dry skin, this is true for elderly persons and also for patients with atopic dermatitis. Therefore, the treatment of the dry skin is one of the most important measures against pruritus. PMID- 16711452 TI - [Autologous fat transplantation in dermatology]. AB - The technique of autologous fat transplantation (lipofilling) allows to correct congenital or acquired abnormalities of the cutaneous relief. Autologous fat lobules aspired in one site are transplanted by injection in the zone to treat. This technique is performed ambulatory under local anesthesia and it is possible to perform long term correction of dermal or hypodermal atrophy of one to few centimetres. PMID- 16711454 TI - [Melanoma prevention in Switzerland: where do we stand?]. AB - Over the last 50 years, skin cancer rates (particularly melanoma) have markedly increased in Caucasian populations. Switzerland, with some 1,600 cases of, and 220 deaths from, malignant melanoma per year has among the highest rates in Europe. This public health issue, affecting relatively young people, has led to primary and secondary nationwide prevention campaigns being carried out for nearly 20 years. Observed changes in sun protection knowledge and attitudes have yet to impact on incidence trend. Early detection has resulted in a large increase in rates of thin melanoma with little change in rates of thick melanoma. Mortality has levelled off and a recent decrease, especially in women, cannot be ruled out. The efficiency of prevention campaigns should soon become more blatant if current efforts persist. PMID- 16711455 TI - [Elementary, Dr. Watson...]. PMID- 16711456 TI - [Mrs. D.'s acute dyspnea]. PMID- 16711457 TI - [What happened on March 13th at the Northwick Park Hospital in London?]. PMID- 16711458 TI - [Subacute bronchitis: which approach should be used with a patient who requests an antibiotic?]. PMID- 16711459 TI - [The good and vibrant appeal of the Cambridge St. John's College]. PMID- 16711460 TI - [Fight against smoking? Xavier Bertrand's smoking room!]. PMID- 16711461 TI - [The disappearance of the subject]. PMID- 16711462 TI - The state of dental health in the north. PMID- 16711463 TI - A political economic history of medical and dental care in Nunavut, Canada. AB - OBJECTIVES: To describe the historical development of medical and dental care in the territory of Nunavut, Canada. STUDY DESIGN: Ethnographic case study using political economy as a mode of explanation. METHODS: Participant observations, document reviews and stakeholder interviews, conducted over a four-year period. RESULTS: There is a clear and now long-term movement from state and professionally controlled health care delivery to Indigenous control over care. CONCLUSIONS: Indigenous groups increasingly hold control over health care delivery through a complex form of management that straddles both public and private organisational spheres. PMID- 16711464 TI - Childbirth among the Canadian Inuit: a review of the clinical and cultural literature. AB - BACKGROUND: This study reviews the historical, anthropological and biomedical literature on childbirth among Canadian Inuit resident in the Canadian Arctic. The modern period is characterised by increased tension as southern intervention replaced traditional birthing with a biomedical model and evacuation to metropolitan hospitals for birth. Inuit concern over the erosion of traditional culture has confronted biomedical concern over perinatal outcomes. Recently, community birthing centres have been established in Nunavik and Nunavut in order to integrate traditional birthing techniques with biomedical support. OBJECTIVES: To review the literature on Inuit childbirth in order to suggest avenues for future research. STUDY DESIGN: Material for this review was gathered through combining library searches, database searches in ANTHROPOLOGYPlus, MEDLINE, CINAHL and Science-Direct, and a bibliographic search through the results. RESULTS: Epidemiological studies of Inuit childbirth are outdated, inconclusive, or inseparable from non-Inuit data. Anthropological studies indicate that evacuation for childbirth has deleterious social and cultural effects and that there is considerable support for traditional communal birthing in combination with biomedical techniques and technology. CONCLUSIONS: Investigation of alternative solutions to maintaining acceptable perinatal outcomes among the Inuit seems desirable. Epidemiological and comparative qualitative studies of perinatal outcomes across the Arctic are needed to reconcile the cultural desirability of communal birthing with claims of its medical feasibility. PMID- 16711465 TI - Fetal and neonatal deaths and congenital anomalies associated with open dumpsites in Alaska Native villages. AB - OBJECTIVE: To determine if women living in Alaska Native villages with open dumpsites ranked as higher hazard have higher rates of adverse pregnancy outcomes than women living in villages with sites that have lower hazard rankings. Adverse pregnancy outcomes examined included fetal and neonatal death and congenital anomalies. STUDY DESIGN: A population-based retrospective cohort study. METHODS: Birth records from 1997-2001 were used to identify the 10,360 eligible infants born to mothers who resided in 197 Alaska Native villages with dumpsite rankings. Exposure variables were derived from hazard rankings of dumpsites. Covariates were obtained from both birth certificate information and village-specific characteristics. RESULTS: Neither crude, nor adjusted estimates detected a statistically significant difference in rates between exposure levels, although adjusted estimates were positive in all congenital anomaly categories, except gastrointestinal defects. Infants born to mothers residing in villages with high hazard dumpsite contents were more likely (RR=4.27; 95% CI: 1.76, 10.36) to have anomalies classified as other defects. Other hazard factors were not significant predictors for any of the adverse outcomes examined. CONCLUSIONS: This is the first study to evaluate fetal and neonatal deaths and congenital anomalies associated with open dumpsites in Alaska Native villages. Problems with the study include a population-based exposure measurement, small sample size, and biases related to birth record information. Future studies should include more comprehensive registries of congenital anomalies. PMID- 16711466 TI - Barriers and supports for healthy eating and physical activity for First Nation youths in northern Canada. AB - OBJECTIVES: To investigate barriers and supports for healthy eating and physical activity in youths in a remote sub-arctic community, Fort Albany First Nation, Ontario, Canada. STUDY DESIGN: A qualitative multi-method participatory approach. METHODS: The study included a purposive convenience sample of two adult (n = 22) and three youths (n = 30; students in grades 6 to 8) focus groups, unstructured one-on-one interviews with adult key informants (n = 7), and a scan of the community environment. Data were coded and analysed by hand and using NVivo software. Hurricane thinking and concept mapping were used to illustrate findings and relationships between concepts. RESULTS: Dominant emerging themes included empowerment, trust, resources, barriers and opportunities, while major sub-themes included food security, cost, accessibility/availability, capacity building, community support, programs/training and the school snack/breakfast program. CONCLUSIONS: Numerous barriers to healthy nutrition and physical activity exist in this community and are possibly similar in other remote communities. Empowerment is a core issue that should be considered in the design of public health interventions for First Nations youths in remote sub-arctic communities. PMID- 16711467 TI - Discharge diagnoses of liver diseases in Nuuk Greenland compared to a Danish county hospital. AB - BACKGROUND: It is a frequently held notion that Inuits/Greenlanders are less prone to develop chronic liver disease than Europeans. High alcohol consumption and chronic viral infection are more frequent in Greenland than in Denmark. STUDY DESIGN: A cross-sectional study to examine the incidence and prevalence of liver discharge diagnosis with focus on cirrhosis among hospital-admitted patients in Greenland and Denmark. METHODS: Register-based ICD-10 discharge diagnoses from Queen Ingrid's Hospital, Greenland, (n = 1072) and Randers Central Hospital, Denmark, (n = 4599) were used to compare the incidence and prevalence of cirrhosis in hospitalised patients during 1998. RESULTS: Five patients (0.47%) in Greenland and 36 (0.78%) in Denmark had a liver discharge diagnosis (OR = 0.67, 95% CI: 0.26-1.72). Two patients (0.19%) in Greenland compared to 25 (0.54%) in Denmark had a cirrhosis associated discharge liver diagnosis (OR = 0.34, 95% CI: 0.08-1.45). The number of newly diagnosed discharged patients was smaller in Greenland, 2 (0.19%), vs. Denmark, 14 (0.30%), (OR = 0.61, 95% CI: 0.14-2.70). CONCLUSION: Discharge liver diagnoses were not more frequent in Greenland than in Denmark--if anything, the hospital prevalence and incidence of liver discharge diagnoses were lower. This may reflect fewer cirrhosis cases in Greenland, and/or a shorter survival time, or lack of follow up. PMID- 16711468 TI - Regional differences in the prevalence of decreased work ability among young employees in Finland. AB - OBJECTIVES: The demographic developments of municipality types differ distinctly. In population-based surveys, the Work Ability Index (WAI) is a useful tool in identifying decreased work ability and high-risk groups. The aim of this study was to examine the prevalence of decreased WAI according to municipal settlements. STUDY DESIGN: Cross-sectional analyses on a birth cohort study base. METHODS: Data on work ability, occupation and marital status were collected by postal questionnaire and in a clinical examination. Areas of residence were obtained from the Population Registration Centre and were delimited using the Finnish rural tripartitioning and urban settlements. Altogether, 3725 employees living in northern Finland or in the Helsinki region during 1997-1998, and working in a wide range of occupations, were selected for the analyses. Analyses of the association between low WAI scores (at or below 36 points), major occupational group and place of residence were carried out separately for men and women by means of logistic regression. RESULTS: At an early stage of the working career, decreased work ability is more likely in rural-type municipalities, especially among women. The regional differences were in close linkage to occupation. CONCLUSIONS: In population-based surveys, the WAI questionnaire measures early signs of decreased work ability and it can be utilized in policy making. PMID- 16711469 TI - Blood pressure and heart rate responses in men exposed to arm and leg cold pressor tests and whole-body cold exposure. AB - OBJECTIVES: Comparison of the effects of different types of cold exposure on blood pressure and heart rate. STUDY DESIGN: Controlled laboratory study. METHODS: Twenty healthy men were exposed to three different types of cold exposure: cold pressor tests for hands and feet, and a 2-h cold air exposure at an ambient temperature of +10 degrees C. RESULTS: All types of cold exposure caused a rise in systolic and diastolic blood pressures. Heart rate increased in the foot cold pressor test. In the cold chamber test, heart rate fell, causing a decrease in the rate:pressure product. No statistically significant correlations were found between the blood pressure responses to cold pressor and cold chamber tests. The increased rate:pressure product observed in the cold pressor tests is indicative of an increased oxygen consumption in the heart muscle. CONCLUSIONS: The measured cardiovascular responses indicate that a sudden local exposure to severe cold would be more stressful than a long lasting, milder exposure to cold, even when the latter is applied to the whole body PMID- 16711470 TI - The usefulness of in vitro models to predict the bioavailability of iron and zinc: a consensus statement from the HarvestPlus expert consultation. AB - A combination of dietary and host-related factors determines iron and zinc absorption, and several in vitro methods have been developed as preliminary screening tools for assessing bioavailability. An expert committee has reviewed evidence for their usefulness and reached a consensus. Dialyzability (with and without simulated digestion) gives some useful information but cannot predict the correct magnitude of response and may sometimes predict the wrong direction of response. Caco-2 cell systems (with and without simulated digestion) have been developed for iron availability, but the magnitude of different effects does not always agree with results obtained in human volunteers, and the data for zinc are too limited to draw conclusions about the validity of the method. Caco-2 methodologies vary significantly between laboratories and require experienced technicians and good quality cell culture facilities to obtain reproducible results. Algorithms can provide semi-quantitative information enabling diets to be classified as high, moderate, or low bioavailability. While in vitro methods can be used to generate ideas and develop hypotheses, they cannot be used alone for important decisions concerning food fortification policy, selection of varieties for plant breeding programs, or for new product development in the food industry. Ultimately human studies are required for such determinations. PMID- 16711471 TI - Dietary and physiological factors that affect the absorption and bioavailability of iron. AB - Iron deficiency, a global health problem, impairs reproductive performance, cognitive development, and work capacity. One proposed strategy to address this problem is the improvement of dietary iron bioavailability. Knowledge of the molecular mechanisms of iron absorption is growing rapidly, with identification of mucosal iron transport and regulatory proteins. Both body iron status and dietary characteristics substantially influence iron absorption, with minimal interaction between these two factors. Iron availability can be regarded mainly as a characteristic of the diet, but comparisons between human studies of iron availability for absorption require normalization for the iron status of the subjects. The dietary characteristics that enhance or inhibit iron absorption from foods have been sensitively and quantitatively determined in human studies employing iron isotopes. People with low iron status can substantially increase their iron absorption from diets with moderate to high availability. But while iron supplementation and fortification trials can effectively increase blood indices of iron status, improvements in dietary availability alone have had minimal influence on such indices within several weeks or months. Plentiful, varied diets are the ultimate resolution to iron deficiency. Without these, more modest food-based approaches to human iron deficiency likely will need to be augmented by dietary iron fortification. PMID- 16711472 TI - Measurements of zinc absorption: application and interpretation in research designed to improve human zinc nutriture. AB - The focus of this paper is on the application of measurements of zinc absorption in human research, especially studies designed to assess the efficacy of intervention strategies to prevent and manage zinc deficiency in populations. Emphasis is given to the measurement of quantities of zinc absorbed rather than restricting investigations to measurements of fractional absorption of zinc. This is especially important when determining absorption of zinc from the diet, whether it be the habitual diet or an intervention diet under evaluation. Moreover, measurements should encompass all meals for a minimum of one day with the exception of some pilot studies. Zinc absorption is primarily via an active saturable transport process into the enterocytes of the proximal small intestine. The relationship between quantity of zinc absorbed and the quantity ingested is best characterized by saturable binding models. When applied to human studies that have sufficient data to examine dose-response relationships, efficiency of absorption is high until approximately 50-60% maximal absorption is achieved, even with moderate phytate intakes. This also coincides approximately with the quantity of absorbed zinc necessary to meet physiologic requirements. Efficiency of absorption with intakes that exceed this level is low or very low. These observations have important practical implications for the design and interpretation of intervention studies to prevent zinc deficiency. They also suggest the potential utility of measurements of the quantity of zinc absorbed when evaluating the zinc status of populations. PMID- 16711473 TI - Methods and options in vitro dialyzability; benefits and limitations. AB - In vitro dialyzability methods involve a two-step digestion process simulating the gastric and intestinal phase, and dialysis through a semi-permeable membrane with a selected molecular weight cut-off. Dialyzable iron/zinc is used as an estimation of available mineral. Final pH adjustment and use of a strict time schedule were found to be critical factors for standardization. In addition the selected cut-off of the dialysis membrane and the method used for iron and zinc determination influence the results. For screening purposes, simple solubility or dialyzability methods seem preferable to the more sophisticated computer controlled gastrointestinal model. This is likely more valuable in studies of different transit times and sites of dialyzability. In vitro solubility/dialyzability methods correlate in most cases with human absorption studies in ranking iron and zinc availability from different meals. Exceptions may be that effects of milk, certain proteins, tea, and organic acids cannot be predicted. The dialyzability methods exclude iron bound to large molecules, which in some cases is available and include iron bound to small molecules, which is not always available. In vitro experiments based on solubility/dialyzability are tools to understand factors that may affect subsequent mineral absorption. PMID- 16711474 TI - Algorithms to assess non-heme iron bioavailability. AB - While sufficient information exists on the effect of individual factors on iron absorption, their net effect in a mixed meal is less well characterized, being dependent on the combination and quantity of the factors present in the meal. Over a period of more than 25 years, several models have been developed to estimate non-heme iron bioavailability, either to assess iron absorption from a meal or iron sufficiency in populations. Initially, a model was developed to calculate iron absorption in individuals with varying iron status that included only enhancers. This model was useful in classifying the diets but has limited value for accurale assessment. Later models were modified and improved by including inhibitors in the calculations. However, some included either phytate or tea but not in combination. The models that included all the factors in calculations assumed their effect was independent and additive rather than interactive, which is an important issue in addressing iron bioavailability. Although some of the models correlated estimated bioavailability with iron status of the population, the accuracy of the estimations is of concern due to lack of quantitative measurements of bioavailability modifiers, inability to consider interactive effects, and the use of non-iron status measurements. Recent research has led to the development of refined models to assess iron bioavailability of complex meals by comprehensively taking into consideration the interactive effect among enhancers and inhibitors. However, the models are based on single-meal studies and their application to whole diets at a population level is not clear. Accurate measurements of dietary factors and independent validation are needed before using these models. To date, no single model is applicable to all diets and additional studies are needed to develop new models to predict bioavailability of whole diets accurately, in addition to addressing dietary adequacy in all populations. PMID- 16711475 TI - Methods and options for estimating iron and zinc bioavailability using Caco-2 cell models: benefits and limitations. AB - Micronutrient deficiencies are prevalent worldwide and have detrimental effects on human health. Complex interactions between micronutrients and other dietary components largely determine micronutrient bioavailability, and understanding these interactions is key to improving micronutrient status. A number of in vitro and in vivo methodologies are available for assessing micronutrient bioavailability. The purpose of this review is to highlight the usefulness of one of the in vitro models, the Caco-2 cell, as a predictive tool for human micronutrient bioavailability. The review focuses on current methods used with the Caco-2 cell line, their benefits and limitations, and the possibilities for the future development of this model. PMID- 16711476 TI - Evidence for the usefulness of in vitro dialyzability, Caco-2 cell models, animal models, and algorithms to predict zinc bioavailability in humans. AB - Low bioavailability of zinc in certain diet types may contribute to zinc deficiency and its consequences in populations. As a result, several experimental models including animal models, in vitro dialyzability models, and Caco-2 cells, have been used to study these factors and estimate their impact on human zinc absorption. For the most part, consistency has been observed between the latter models and human absorption studies to identify factors that enhance or inhibit zinc bioavailability. However, dialyzability methods are limited to modeling luminal interactions among the factors as they affect zinc availability while Caco-2 cells can model luminal effects and uptake by intestinal cells. Neither animal nor in vitro methods can predict the magnitude of zinc absorption at the level of the human organism. Caco-2 cells will be useful models for understanding the mechanisms of intestinal zinc absorption. The in vitro methods are also limited to modeling absorption and the interactions that occur. Algorithms to estimate zinc absorption, based on dietary content of zinc absorption modifiers, have been derived from human studies. An algorithm derived from studies of zinc retention from radioactive zinc-labeled test meals underestimates zinc absorption compared to that derived from measurement of true zinc absorption from total diets using isotopic tracer methods. Based on the latter, phytate appears to be the only major inhibitor of zinc absorption from typical diets. Ultimately, population-based studies are needed to determine the impact of dietary factors that modulate zinc absorption on the adequacy of zinc status. PMID- 16711477 TI - The precision of in vitro methods and algorithms for predicting the bioavailability of dietary iron. AB - Three factors determine how much iron will be absorbed from a meal. They are the physiological mechanisms that regulate uptake by and transfer through the enterocytes in the upper small intestine, the quantity of iron in the meal, and its availability to the cellular iron transporters. Established methods exist for predicting the effect of physiological regulation and for measuring or estimating meal iron content. Three approaches to estimating bioavailability have been advocated. Two are in vitro screening procedures: measurement of dialyzable iron and Caco-2 cell uptake, both carried out after in vitro simulated gastric and pancreatic digestion. The third is the use of algorithms based on the predicted effects of specific meal components on absorption derived from isotopic studies in human volunteers. The in vitro procedures have been very useful for identifying and characterizing factors that affect non-heme iron absorption, but direct comparisons between absorption predicted from the in vitro tests and measurements in human volunteers have only been made in a limited number of published studies. The available data indicate that dialysis and Caco-2 cell uptake are useful for ranking meals and single food items in terms of predicted iron bioavailability, but may not reflect the magnitudes of the effects of factors that influence absorption accurately. Algorithms based on estimates of the amounts of heme iron and of enhancers and inhibitors of non-heme iron absorption in foods make it possible to classify meals or diets as being of high, medium, or low bioavailability. The precision with which meal iron bioavailability can be predicted in a population, for which a specific algorithm has been developed, is improved by measuring the content of the most important enhancers and inhibitors. However, the accuracy of such predictions appears to be much lower when the algorithm is applied to meals eaten by different populations. PMID- 16711478 TI - [Rheumatic patients have not only joint disease]. PMID- 16711480 TI - [Wart removed, foot lost]. PMID- 16711479 TI - [When are "biologics" indicated?]. PMID- 16711481 TI - [Occupational dermatoses. Here you must help entirely not in a bureaucratic way]. PMID- 16711482 TI - [Breakthrough in gene therapy? Interview by Friederike Klein]. PMID- 16711483 TI - [Soft tissue tumors are mostly benign, but malignant sarcoma should not be overlooked!]. PMID- 16711484 TI - [Soft tissue sarcoma--epidemiology, diagnosis and treatment]. AB - On account of the considerable variability of the clinical situation, treatment of soft tissue sarcoma requires an individually oriented multimodal approach. In the case of patients with superficial low-grade tumors measuring less than 5 cm in diameter, resection alone is usually adequate. In the event of medium-grade lesions, resection with negative margins, resection in combination with radiotherapy achieves excellent local control rates associated with an overall survival rate of almost 80%. In patients with high-grade sarcomas measuring more than 5 cm in diameter, local control can be achieved with resection and radiotherapy, although every second such patient develops metastases. For patients with local recurrence, further resection should be considered/performed. Radiotherapy is all the more effective, the smaller the postoperative tumor cell burden. PMID- 16711486 TI - [Diagnosis of persistent rhinitis]. AB - A total of 9923 patients with persistent rhinitis were characterized on the basis of their symptoms and in vitro data. The most bothersome symptom in these patients was nasal obstruction. "Post-nasal drip" proved to be untypical for allergic rhinitis. In all the diseases investigated, involvement of eosinophilic granulocytes and mast cells was found to vary. In addition to endoscopy and imaging procedures, the determination of selective in vivo and in vitro parameters represents an essential part of the diagnostic work-up of persistent rhinitis, which thus proves to be a domain of the allergological rhinologist. PMID- 16711485 TI - [Soft tissue sarcoma of the extremities: current state of the art of adjuvant therapy]. AB - Today, most sarcoma patients can be spared an amputation through the use of adjuvant radiotherapy. Treatment by an experienced multidisciplinary team offers the best chance of achieving permanent tumor control. Histopathologically-free resection margins are of the greatest importance. The indication for radiotherapy is determined by the recurrence risk profile of the individual patient. In addition to the well-proven postoperative irradiation, neoadjuvant radiotherapy is also successful. In the event of an unfavorably sited tumor, intra-operative irradiation can be applied in combination with either form. Patients with large G3 tumors can be given adjuvant chemotherapy to reduce the risk of distant metastases. On account of its appreciable toxicity, however, it should be reserved for patients younger than 65 in a good state of health. PMID- 16711487 TI - [What is your diabetic patient allowed to eat]. AB - The Expert Group Diabetes and Nutrition of the European Diabetes Association has compiled evidence-based recommendations for the treatment and prevention of diabetes mellitus. These recommendations have been coordinated with all the major relevant societies in Germany. This has enabled a uniform approach to the dietary treatment of persons with diabetes that is supported by the German Diabetes Association, the German Obesity Association, the German Association of Nutritional Medicine and the German Association for Nutrition. The scientific background to the dietary recommendations have also been incorporated in a nutrition training programme and a patient information guide with the aim of promoting countrywide implementation. PMID- 16711488 TI - [Chronic non-malignant pain. Opioids are also suitable for long-term treatment]. PMID- 16711489 TI - [From when does an ultrasonography equipment pay for itself?]. PMID- 16711491 TI - [Total savings effect up to 3 billions Euro]. PMID- 16711490 TI - [Tamsulosin in the sustained-release capsule -- continuously effective and economical]. PMID- 16711492 TI - [Creatinine increase as accidental finding]. PMID- 16711493 TI - [Gloomy facial expression, giant hands]. PMID- 16711494 TI - [Continuous perineural blocks for postoperative analgesia in ambulatory care]. PMID- 16711495 TI - [Postpartum and early postoperative anemia after gynecological surgery: treatment with intravenous iron]. AB - OBJECTIVE: To assess the efficacy and safety of treatment with intravenous iron for postpartum anemia or postoperative anemia after gynecological surgery. PATIENTS AND METHODS: A prospective study enrolling patients entering our recovery care unit from June through December 2004 with hemoglobin levels less than 10 g x dL(-1) after vaginal childbirth or cesarean section or after another form of gynecological surgery. Three 200 mg doses of intravenous iron sucrose (Venofer) were administered on consecutive days. Fifteen days after the last dose, the patient came for follow-up tests and was asked about side effects. The results were analyzed with a Student t test for matched samples. RESULTS: A total of 250 obstetric and 52 gynecological surgery patients were enrolled; 156 and 33 completed the study in each group, respectively. Hemoglobin increased after treatment by 3.2 g x dL(-1) in the obstetric patients and by 2.7 g x dL(-1) in patients who underwent gynecological surgery. The increase was significant in both groups (P<0.001); the 95% confidence interval was 2.918-3.519 for the obstetric patients and 2.220-3.071 for the gynecological surgery patients. The incidence of side effects was low (13 obstetric patients and 1 gynecological surgery patient). Most side effects were related to pain at the injection site (in 12 of the 14 women). CONCLUSIONS: Intravenous iron sucrose is safe and effective for treating puerperal anemia and following gynecological surgery. The low incidence of serious side effects and the rapid recovery of hemoglobin levels make this a safe, effective drug for treating anemia. PMID- 16711496 TI - [Continuous versus single-dose sciatic nerve block to complement a femoral block after total knee replacement surgery: a randomized clinical trial]. AB - INTRODUCTION: Performing a sciatic nerve block to complement a continuous femoral nerve block for analgesia after total knee arthroplasty is a subject of controversy. We compared the efficacy of a continuous sciatic nerve block to that of a single-dose block of the same nerve combined with a continuous femoral nerve block. MATERIAL AND METHODS: Patients received a continuous femoral nerve block by infusion of 0.2% ropivacaine, 0.4 mL x kg(-1), plus patient controlled analgesia and were randomized to receive a sciatic nerve block either by continuous infusion (0.5% ropivacaine, 20 mL, plus continuous infusion of 5 mh( 1)) or by a single 20 mL dose of 0.5% ropivacaine. The stimulating catheters remained indwelling for 72 hours after the operation. The main outcome measure was assessment of postoperative pain on a verbal numerical scale from 0 (no pain) to 10 (greatest pain) at rest and upon movement. RESULTS: The pain assessments upon movement of the operated joint were significantly greater after 24 hours in the group receiving a single dose of ropivacaine to block the sciatic nerve. The back of the knee was most often named as the location of pain. No adverse effects related to the analgesic technique were recorded, and the level of satisfaction was higher among patients receiving the continuous sciatic nerve block. CONCLUSION: Twenty-four hours after total knee replacement surgery, better analgesia was achieved with a continuous sciatic nerve block than with a single dose block combined with a continuous femoral nerve block. PMID- 16711498 TI - [Techniques to block the sciatic nerve by a lateral approach through the popliteal fossa]. AB - Lateral approaches to the sciatic nerve through the popliteal fossa have recently been described as useful for providing adequate anesthesia and postoperative analgesia for foot and ankle surgery. Numerous publications have appeared on the approach in recent years, proposing new anatomical landmarks to facilitate location of the nerve, reduce the rate of complications, and increase the rate of success. When the lateral popliteal approach has been compared to other approaches to the sciatic nerve, similar success rates have been observed. However, when this technique is used certain factors must be borne in mind because they can influence both latency time and success. This review describes the lateral popliteal approach, its main variations, the factors that can affect latency time or success, and the possibility of providing continuous analgesia. We also sought to compare this approach to other techniques for blocking the sciatic nerve. PMID- 16711497 TI - [Oral analgesia versus posterior tibial nerve block for postoperative pain in ambulatory open hallux valgus surgery: a randomized clinical trial]. AB - INTRODUCTION: This trial assessed the safety and efficacy of a continuous posterior tibial nerve block in the ankle provided in the patient's home by elastomeric pump infusion of 0.375% ropivacaine after ambulatory hallux valgus surgery. MATERIAL AND METHODS: Patients were randomized to 2 groups of 20 each to receive either the conventional oral analgesia prescribed by our team after outpatient surgery (metamizole 575 mg/6 h p.o.) or perineural analgesia with a continuous infusion of 5 mL x h(-1) of 0.375% ropivacaine in the posterior tibial nerve. Surgery was performed under hyperbaric spinal anesthesia with mepivacaine and an injection of 0.25% bupivacaine into the joint. Both groups also received 50 mg/8 h p.o. of tramadol as rescue analgesia. Assessment during visits by the home care team 12, 24, and 48 hours after surgery included the following variables: pain on a visual analog scale (VAS, 0-10), sleep quality, need for rescue analgesia, acceptance of the technique, side effects and adverse events. Descriptive statistics were calculated and comparisons were performed with the Mann-Whitney U test; sleep quality and need for rescue analgesia were compared by applying the chi2 statistic with a test of linear trend. RESULTS: The perineural analgesia group had significantly lower VAS scores at 4, 12, and 24 hours and less need for rescue analgesia. No differences in sleep quality were found (P0.07). The incidence of side effects did not differ, and there were no readmissions. The patients expressed a high level of acceptance of the technique. CONCLUSION: Continuous perineural analgesia in the home setting was found to be effective and safe in our patients. PMID- 16711499 TI - [Complications of peripheral nerve blocks]. AB - Recent years have brought renewed interest in peripheral nerve blocks as new drugs and materials have become available and as the advantages of such blocks over other anesthetic and analgesic techniques have been demonstrated. We conducted a literature search in PubMed, the Cochrane Library Plus (Spanish version), and Healthgate for the period January 1, 2000 through October 15, 2005 using the key words "complication and peripheral nerve block," "complication and peripheral block anaesthesia," and "complication and plexus anaesthesia." Totals of 178, 109, and 145 titles were obtained with each portal. When the search was limited to reviews, clinical trials, and randomized controlled trials, the numbers of articles from each portal became 11, 20, and 10, respectively, for the first string of key words; 0, 8 and 6, respectively, for the second string; and 10, 40 and 16 for the third. A detailed analysis of the results revealed 2 reviews and 1 clinical trial in which complications of peripheral nerve blocks figured in the main topic. Most of these publications agree that the overall incidence of complications related to these techniques is low. If practice guidelines and algorithms from experts are followed the incidence can be reduced. The aim of this review is to give an updated report of complications related to peripheral nerve blocks. PMID- 16711500 TI - [Locoregional anesthesia of the penis: focus on the blockade of the cavernous body]. PMID- 16711501 TI - [Anesthetic management of a boy with sialidosis]. AB - Sialidosis is an autosomal recessive disease caused by neuraminidase deficiency, which causes material rich in sialic acid to accumulate in various tissues and organs, leading to morphological and functional changes. Mainly the osteoarticular, cardiorespiratory, and central nervous systems are affected. We report the case of a 14-year-old boy diagnosed with sialidosis type 2 who was proposed for spinal column arthrodesis to treat severe thoracolumbar scoliosis. Physical examination revealed pectus carinatum; facial deformity and macroglossia; hypertelorism and frontal bossing; short neck; atlantoaxial subluxation; severe arthrogryposis of elbows, knees and wrists, coxa valga, and club foot. Relevant medical history included moderately severe aortic valve insufficiency and myoclonic epilepsy. Surgery was performed under balanced anesthesia. Awakening was delayed and the patient was transferred to the intensive care unit under mechanical ventilation. Anesthetic problems that can present in patients with such deformities include difficult airway, complicated management of associated cardiopathy, and difficult weaning from mechanical ventilation. PMID- 16711502 TI - [Tracheal laceration after intubation and application of continuous airflow through the outer cuff of the tracheal tube]. AB - We report the case of a man diagnosed with cervical spinal stenosis who underwent a C3-C6 bilateral laminectomy. In the immediate postoperative period he developed subcutaneous emphysema and pneumomediastinum caused by tracheal laceration. Continuous airflow (1 L x min(-1)) through the outer cuff was applied during the operation to compensate for air leak when loss of balloon cuff pressure was detected. PMID- 16711503 TI - [Accidental dural puncture during epidural injection of corticosteroids: a different approach?]. AB - We report 6 cases diagnosed with accidental dural puncture after epidural injection of corticosteroids for low back pain. All the patients reported postdural puncture headache during their stay in the postanesthetic recovery unit. For 3 patients, pain resolved with treatment given in the recovery unit. Two other patients also required mild analgesics for 1 week. In the last patient, a blood patch was used to treat incapacitating headache 22 days after the epidural procedure and mild analgesics were needed for 4 more weeks. It is important to establish a protocol for treating postdural puncture headache in pain clinics to facilitate decision making. Good physician-patient communication is necessary to avoid refusals for permission for other epidural techniques and to facilitate management of symptoms. PMID- 16711504 TI - [Anesthesia and vitamin B12]. PMID- 16711505 TI - [Activated recombinant factor VII for bleeding after a kidney transplant: reflections]. PMID- 16711506 TI - [Intraneural sciatic nerve puncture in the popliteal fossa demonstrated by ultrasound]. PMID- 16711507 TI - On the clinical usefulness of the free-to-total prostate-specific antigen ratio. AB - The free-to-total prostate-specific antigen ratio (F/T PSA) is associated with the presence of prostate cancer and is thus used as an indicator for suspicion of prostate cancer and as a determinant for biopsy. We reviewed a recent retrospective series of 966 consecutive prostate biopsies where F/T PSA was blindly determined and did not influence biopsy indication. We simulated the association of F/T PSA with biopsy outcome and its impact as a biopsy determinant. When adopting an F/T PSA cutoff of 10%, 13%, 16% or 20% among random sextant biopsies in the 4-10 ng/mL total PSA range, the sensitivity was 15%, 37%, 55% and 72% and the specificity 89%, 80%, 64% and 44%, respectively. Using F/T PSA as a biopsy determinant, from 1.7 to 2.6 cancer biopsies would have been delayed to avoid 10 benign biopsies. As this balance is not acceptable, F/T PSA has no role as a biopsy indicator and its clinical use is questionable. PMID- 16711508 TI - Levels of tissue inhibitor of metalloproteinases 1 in plasma and urine from patients with bladder cancer. AB - AIM: To assess the potential use of plasma and urine levels of tissue inhibitor of metalloproteinases 1 (TIMP-1) in urothelial cancer. METHODS: TIMP-1 levels were determined in urine and plasma from healthy donors (n=26), patients with bacterial bladder infection (n=24), urothelial bladder adenoma (n=3) or adenocarcinoma (n=7). RESULTS: Free and total TIMP-1 in plasma were weakly but significantly correlated with age; urinary TIMP-1 was not. A strong correlation between free and total TIMP-1 in plasma was observed, with an average ratio of 0.85. No correlation between total TIMP-1 in urine and plasma was found (p=0.55). No significant differences in free or total TIMP-1 in plasma were found between healthy individuals, patients with cystitis or bladder cancer (p=0.4). Urinary TIMP-1 levels were significantly increased in patients with cystitis (p=0.001). No apparent differences in TIMP-1 levels were found in patients with bladder cancer at different stages. CONCLUSION: Our previous observation of a weak but significant correlation between plasma TIMP-1 and age was confirmed. Likewise, an association between free and total TIMP-1 in plasma with a ratio of 0.85 was established. No correlation between plasma and urine TIMP-1 was found. Measurement of TIMP-1 in plasma and/or urine is apparently not useful for the identification of bladder cancer. PMID- 16711509 TI - Phosphorylated Akt1 in human breast cancer measured by direct sandwich enzyme linked immunosorbent assay: Correlation with clinicopathological features and tumor VEGF-signaling system component levels. AB - Protein kinase B (Akt) plays a major role in the regulation of breast cancer growth, survival, hormone, drug and radiosensitivity, but the clinical value of its expression and activation in human tumors is unclear. Activated Akt1 (pAkt1) expression was quantified in a series of 46 breast cancer and adjacent mammary gland samples by a direct Path-Scan PhosphoAkt1 (Ser473) sandwich ELISA kit. VEGF, sVEGFR1 and sVEGFR2 levels were measured simultaneously by standard ELISA kits. Forty-nine percent of the tumors had an increased pAkt1 level as compared to adjacent tissue. pAkt1 levels were significantly higher in stage IIb than in stage I-IIa tumors. The frequency of pAkt1 elevation was positively associated with tumor size and malignancy grade. pAkt1 was also twice as frequently increased in PgR-negative as in PgR-positive tumors, while its mean level was significantly higher in ER-positive than in ER-negative tumors. VEGF, sVEGFR1 and sVEGFR2 were increased in 73-85% of the tumors, but no associations with most clinicobiological factors and pAkt1 level were found. In conclusion, activation of Akt1 is not associated with VEGF signaling protein expression in breast cancer but is related to tumor size, grade of malignancy, and steroid receptor status. PMID- 16711510 TI - Reliability and discriminant validity of HER2 gene quantification and chromosome 17 aneusomy analysis by real-time PCR in primary breast cancer. AB - There is an increasing demand for the evaluation of HER2 status in breast cancer. In this study, sections from fixed tissues and triton extracts of tissue homogenates were obtained from 163 malignant breast tumors and analyzed in parallel using immunohistochemistry combined with fluorescence in situ hybridization, as gold standard tests, and an ELISA test (c-erbB2/c-neu Rapid Format ELISA, Oncogene Research Products, USA). Tumor DNA was employed to evaluate two quantitative PCR methods: the HER2/neu DNA Quantification Kit (Roche Diagnostics GmbH, Germany), which uses the gastrin chromosome 17 reference gene, and our recently developed Oncolab qPCR assay, where both a chromosome 17 gene (somatostatin receptor type II (SSTR2)) and a non-chromosome 17 reference gene (glyceraldehyde-3-phosphate deshydrogenase (GAPDH)) were used to detect an increase in HER2 gene copy number and to evaluate the aneusomy of chromosome 17, respectively. By IHC/FISH and ELISA, HER2 was overexpressed in 27 (16.6%) and 24 (14.7%) samples, respectively. With the Roche and Oncolab qPCR assays, 29 (17.8%) samples showed a ratio of HER2/gastrin > or = 2.0 and 26 (16.0%) showed a ratio of HER2/SSTR2 > or = 2.0, respectively. In samples presenting HER2/SSTR2 <2.0 and HER2/GAPDH > or = 2.0, which was indicative of a chromosome 17 polysomy, we observed a modest increase in HER2 protein expression. Complete agreement between the four methods for HER2 status determination was obtained for 154 (94.5%) samples. Overall, these results demonstrate that quantitative PCR is a reliable method for analyzing HER2 status and chromosome 17 polysomy. PMID- 16711511 TI - Simultaneous quantitative detection of relevant biomarkers in breast cancer by quantitative real-time PCR. AB - The assessment of ERa, PgR and HER2 status is routinely performed today to determine the endocrine responsiveness of breast cancer samples. Such determination is usually accomplished by means of immunohistochemistry and in case of HER2 amplification by means of fluorescent in situ hybridization (FISH). The analysis of these markers can be improved by simultaneous measurements using quantitative real-time PCR (Qrt-PCR). In this study we compared Qrt-PCR results for the assessment of mRNA levels of ERa, PgR, and the members of the human epidermal growth factor receptor family, HER1, HER2, HER3 and HER4. The results were obtained in two independent laboratories using two different methods, SYBR Green I and TaqMan probes, and different primers. By linear regression we demonstrated a good concordance for all six markers. The quantitative mRNA expression levels of ERa, PgR and HER2 also strongly correlated with the respective quantitative protein expression levels prospectively detected by EIA in both laboratories. In addition, HER2 mRNA expression levels correlated well with gene amplification detected by FISH in the same biopsies. Our results indicate that both Qrt-PCR methods were robust and sensitive tools for routine diagnostics and consistent with standard methodologies. The developed simultaneous assessment of several biomarkers is fast and labor effective and allows optimization of the clinical decision-making process in breast cancer tissue and/or core biopsies. PMID- 16711512 TI - Arginase as a useful factor for the diagnosis of colorectal cancer liver metastases. AB - The present work is a continuation of studies on arginase as a marker in the diagnosis of colorectal cancer liver metastases (CRCLM). The purpose of the study was the evaluation of the arginase test in comparison with other colorectal cancer tests such as CEA, CA 19-9 and biochemical markers of liver function such as aspartate aminotransferase (AST) and alanine aminotransferase (ALT). The studies were conducted on blood serum from 85 patients with CRCLM obtained one to two days before tumor resection. The control group comprised 140 healthy blood donors and 81 patients with various non-malignant gastrointestinal diseases. Raised arginase activity was observed in serum of 85% of CRCLM patients, whereas elevated levels of CEA and CA 19-9 were found in 63% and 42% of patients, respectively. The combination of CEA or CA 19-9 with the arginase assay improved their sensitivity, but the sensitivity of the combined parameters was not higher than that of the arginase test itself. AST and ALT activities were increased in about 30% of CRCLM patients. The specificity of the arginase test calculated for 221 control subjects was 76%. It can thus be concluded that the determination of serum arginase activity can be helpful in the diagnosis of patients with colorectal cancer liver metastases. PMID- 16711514 TI - Association of TP53 mutation, p53 overexpression, and p53 codon 72 polymorphism with susceptibility to apoptosis in adult patients with diffuse astrocytomas. AB - Clarification of TP53 alterations is important to understand the mechanisms underlying the development of diffuse astrocytomas. It has been suggested that the alleles of TP53 at codon 72 differ in their ability to induce apoptosis in human cancers. The aim of this study was to analyze the possible association of TP53 mutation, p53 overexpression, and p53 codon 72 polymorphism with susceptibility to apoptosis in adult Brazilian patients with diffuse astrocytomas. We analyzed 56 surgical specimens of diffuse astrocytomas for alterations of TP53, using polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) direct sequencing. p53 and cleaved caspase 3 protein expression were assessed by immunohistochemistry. We found TP53 mutations in 19.6% (11 out of 56) of tumors tested, with the lowest mutation rate found in the cases of glioblastomas (8.8%) (p = 0.03). Only 16.1% of tumors tested showed cleaved caspase 3-positive staining, demonstrating that apoptosis is very inhibited in these tumors. All tumors having TP53 mutation and p53 accumulation had no expression of cleaved caspase 3. Additionally, no association was observed in tumors having proline and arginine alleles and expression of cleaved caspase 3. We concluded that clarification of the TP53 alterations allows a better understanding of the mechanisms involved in the progression of diffuse astrocytomas, and the allele status at codon 72 was not associated with apoptosis in these tumors. PMID- 16711513 TI - Clinical utility of hyperglycosylated hCG in serum taken before hydatidiform mole evacuation to predict persistent trophoblastic disease. AB - OBJECTIVE: Human chorionic gonadotropin (hCG) is widely used in the management of hydatidiform mole and persistent trophoblastic disease (PTD). Studies on hyperglycosylated human chorionic gonadotropin (invasive trophoblast antigen, ITA) in PTD are limited. In serum samples taken before evacuation of molar pregnancies we measured the concentrations of free hCG beta-subunit (free hCGbeta), "total" hCG (hCG+hCGbeta) and ITA, and determined whether ITA, the two other hCG analytes, or the calculated ratios of hCGbeta/hCG+hCGbeta, hCGbeta/ITA and hCG+hCGbeta/ITA could predict the later development of PTD. DESIGN: A retrospective study based on blood specimens collected in the Dutch Central Registry for Hydatidiform Moles. The study group comprised 97 patients with hydatidiform moles who did not develop PTD after mole evacuation and 33 patients who did develop PTD. METHODS: Serum samples from 130 patients with hydatidiform mole with or without PTD were assayed using specific (radio)immunoassays for free hCGbeta, total hCG, and ITA. From these analytes we also calculated the ratios hCGbeta/hCG+hCGbeta, hCGbeta/ITA, and hCG+hCGbeta/ITA. To predict the development of PTD from these analytes and parameters we performed receiver-operating characteristic (ROC) curve analysis, resulting in areas under the curve (AUCs) that represented the diagnostic accuracy which was rated in a range from excellent (AUC >0.9 or <0.1) to poor (AUC 0.4-0.6). RESULTS: The diagnostic accuracy of ITA was moderate (0.618) and not different from that of free hCGbeta (0.610) and hCG+hCGbeta (0.622). CONCLUSIONS: ITA as well as the other analytes and parameters in serum taken prior to evacuation from patients with molar pregnancies cannot be used to predict the subsequent development of persistent trophoblastic disease. PMID- 16711515 TI - Chromogranin A blood concentrations in relation to those of prolactin in hormone refractory metastatic prostate cancer: possible existence of two independent mechanisms responsible for endocrine resistance. PMID- 16711516 TI - Measurement of the effect of a plant-based diet on reactive oxygen metabolite production using the d-Roms test is invalid. PMID- 16711517 TI - Construction success: it's all in the planning. HFMA's annual real estate, design, and construction review. PMID- 16711518 TI - Consumer-directed care: what will it mean to your business office? AB - As healthcare costs continue to escalate, consumer-directed care is emerging as a means for patients to gain greater control over their healthcare decisions- including where their dollars are going. While no one knows what effect this trend ultimately will have, HFMA, with sponsorship from McKesson Corporation, convened a group of five senior financial executives this spring in San Antonio to gain greater perspective on changes the business office is likely to face. This "visioning" session serves to spark others to examine potential repercussions of the trend within their own markets and actions needed to prepare for what may come. PMID- 16711519 TI - Unsafe at any cost: mediocre U.S. health care. PMID- 16711520 TI - The balancing role of balance billing. AB - A recent court case, Prospect Medical Group, Inc. v. Northridge Emergency Medical Group et al., has ruled that emergency physicians are allowed to balance bill health plan subscribers when the plan itself underpays the providers. Partly at issue was whether the providers' contract with the health plan was only implied. The court's advice: Put all contracts in writing. PMID- 16711521 TI - Jurisdictional issues related to PRRB appeals. AB - Protecting your right to appeal a final determination on program reimbursement requires a knowledge of the mandatory criteria that must be met as well as the jurisdictional issues related to Provider Reimbursement Review Board appeals. Common jurisdictional issues include: Final determination. Provider satisfaction with a final determination. Reopenings. Adding issues to an appeal. Expedited judicial review. PMID- 16711522 TI - The big squeeze in search of a sustainable future for suburban community hospitals. AB - Suburban community hospitals face increasing competition from two sides-from larger regional providers and freestanding diagnostic and treatment centers. To effectively meet this challenge, these hospitals require rigorous integrated strategic financial planning. The hospitals should take a close look at their key differentiation strategies with respect to operational efficiency, performance and quality, program and service mix, market focus, physician staff alignment, and system linkage. PMID- 16711523 TI - A CFO's guide to the marketing galaxy. AB - The following questions will help you evaluate your organization's marketing willingness: What is your organization's risk tolerance? Is your organization best suited for acquisition or for retention? Does your organization know the value of a patient? Does your organization have the right marketing infrastructure? PMID- 16711524 TI - Step up to the step-down method. AB - CFOs can use the Medicare cost report step-down method to allocate costs by service line and then develop an income matrix. With an income matrix and tiered expenses by service areas, CFOs can direct corrective action to improve financial performance. The cost data derived from the step-down method can also be used to set prices and to negotiate third-party contracts. PMID- 16711525 TI - Planning your cancer service line three steps to success. AB - One Midwest health system found that by using a three-step approach to bolster the success of its lung cancer program, the program exceeded patient volume targets within its first year of operation. The program now serves as a business model for other investment opportunities in site-specific cancer programs at the health system. PMID- 16711526 TI - Staying on course with strategic metrics. AB - Providers need to understand the various types of metrics, the importance of each type, and how to use them effectively. Providers should use strategic metrics to monitor implementation of their organization's strategic plan. To overcome obstacles to successful use of strategic metrics, providers should communicate the organization's strategic intent, establish accountability for achieving the targets, and ensure buy-in through review of metrics and involvement of appropriate people. PMID- 16711527 TI - 5 facility planning mistakes (and how to avoid them). AB - To avoid some common facility planning mistakes, healthcare financial executives should: Solve problems rather than symptoms. Start with population needs rather than perceived space needs. Focus on accuracy rather than precision. Focus on room quantity rather than room size. Base planning on function rather than preference. PMID- 16711528 TI - Can you bank on philanthropy? AB - CFOs can help to establish a culture of giving within their organizations by: Helping to build a culture of philanthropy within the hospital. Including fund raising performance on the hospital's dashboard measurements. Learning fund raising language and etiquette to enhance interaction with potential donors. Writing articles for hospital newsletters and local newspapers, explaining why philanthropy is so vital to the future of health care in the community. PMID- 16711529 TI - Diversity's next challenge. AB - A healthcare organization's diversity initiative will help the organization identify and eliminate barriers that limit employee and patient satisfaction. The way services are being provided in most hospitals needs to be modified to reflect the changing demographics of the populations being served. Creating a culturally sensitive healthcare organization may require changing recruiting strategies or entering new networks. PMID- 16711530 TI - The monthly reserve valuation process. PMID- 16711531 TI - HIPAA enforcement rule will more teeth equal bigger bite? PMID- 16711532 TI - Who's in charge of your growth agenda? PMID- 16711533 TI - The balance sheet: a snapshot of your financial health. PMID- 16711534 TI - Analyzing the key indicators for total utilization and departmental volumes. PMID- 16711535 TI - Food-dependent exercise-induced anaphylaxis--update and current data. AB - Exercise-induced anaphylaxis (EIA) is defined as the onset of allergic symptoms during, or immediately after, exercise, the clinical signs being various degrees of urticaria, angioedema, respiratory and gastrointestinal signs and even anaphylactic shock. Food-dependent exercise-induced anaphylaxis (FDEIA) introduces food in the syndrome and is revealed by a chronological sequence in which food intake, followed by exercise, induces symptoms after a varying period. When the food intake and the exercise are independent of each other, there are no symptoms. FDEIA is not very frequent. Identifying the culprit food allergen depends on the patient's eating habits. Crustaceans and wheat flour are the two commonest but others foods can be implicated. The patho-physiology of FDEIA has not been clearly established but it appears to result from degranulation of mast cells. As with food allergy, FDEIA diagnosis is based on interview, skin and biological tests and challenge. For the clinical signs of allergy, antihistamines, corticosteroids and epinephrine may be administered. Prophylaxis aims to prevent a recurrence; the patient should be given an emergency kit to deal with any recurrent episode. After the food allergen has been identified, it should be avoided for at least 4 to 5 hours before any exercise. PMID- 16711536 TI - The risk of systemic reactions to skin prick-tests using food allergens: CICBAA data and literature review. AB - Prick-tests to foods are usually carried out as the first step in the diagnosis of food allergy. Severe anaphylaxis accounts for 4.9 % of allergies in children and occurs more frequently in adults, raising the possibility of systemic reactions to prick-tests in highly sensitized people. Several studies published in the literature have used commercial extracts. As for airborne allergens, concentrations causing a skin reaction of 15 mm do not present a risk of systemic reactions. Prick-tests to native foods--prick-in-prick tests--have been less extensively studied. The CICBAA1 data, from 1,138 food allergic patients of all ages, cover 34,905 prick-in-prick tests to foods. The wheal of these prick-tests has been regulary registered. The risk of systemic reactions can be evaluated at 0.008 %. There were no severe reactions and anti-histamine and corticosteroid therapy were sufficient. These results are similar to those of the large study in 2000 carried out by Devenney in neonates (0.005%). A review of the literature reveals only a few severe reactions in adults. The authors draw attention to the necessary precautions: temporary contra-indication for skin prick-tests in children and adults with grade 3 or 4 asthma, with particular attention to such foods as all kinds of nuts, fish, etc. PMID- 16711537 TI - Cell typing of the scabetic lesion and its clinical correlation. AB - Scabies is a pruritic dermatosis due to infestation by a mite that stimulates both humoral and cellular immune responses causing cutaneous lesions as well as pruritus. Changes in immune response pattern with treatment are associated with improvement in symptoms and clinical cure. In this study, we analysed the cellular infiltrate types and patterns in lesions of Scabies. Treated patients were divided into 2 groups as those with and without pruritus. In the pre treatment cases, there was a high incidence of T3, T4, T6 and T8 infiltration in the lesions, with T4 greater than T8. Following treatment, it was noted that T8 density was significantly increased in patients who had resolution of symptoms. It can therefore be concluded that T4 cell dominance is the cause of persistant itching and T8 increase leads to improvement in the pruritus. PMID- 16711538 TI - Wheat flour allergy: an entire diagnostic tool for complex allergy. AB - Wheat proteins are involved in respiratory allergy, contact allergy and food allergy. Wheat allergens involve in these pathologies are well-known. However, establishment of wheat allergy diagnostic can be sometimes difficult on account of the complex allergenic composition of skin prick test (SPT) solutions of wheat flour. Therefore, we have studied specific IgE reactivity from patient sera with wheat food allergy, and characterized allergenic composition of wheat SPT solutions by specific antibodies directed to wheat allergens. The results showed that 20 of the 25 sera analyzed contained specific IgE to at least one wheat protein fraction. Among positive sera, 75% have specific IgE to water/salt soluble fraction, 85% to native gluten fractions and 65% to wheat isolate fraction. The results showed also that SPT solutions of wheat flour contained major food allergens from each allergenic fraction. These results highlighted the importance of using fractions, which constitute the whole wheat allergenic pattern, during specific IgE reactivity analyses. Moreover, we have observed that wheat isolate extract (results of food industrial process) contained not only modified allergens (neo-allergens) involve of specific food allergy to wheat isolate but also some native allergens involve in wheat food allergy. Thus, these results showed the importance to use, for wheat in vivo diagnosis together wheat SPT solutions (gluten extract and wheat isolate) in order to differentiate wheat food allergy to specific wheat isolate allergy. PMID- 16711539 TI - A comparative study between viral isolation and indirect immunofluorescence in the diagnosis of herpes simplex virus. AB - Fifty patients with oral ulcers were studied clinically and investigated for the detections of Herpes Simplex Virus (HSV) through virus isolation from their lesions (vesicles and ulcers) and detection of the presence of antiviral antibodies (both, IgM and IgG) in their sera using the indirect immunofluorescene (IIF) technique. The results of this study proved that virus isolation is the most reliable method for diagnosis, though the use of antibody serological tests could be a useful adjunct to virus isolation in situations where a rapid laboratory diagnosis is needed. Oral Herpes Simplex virus infection can be viewed, in the main, as a trivial disorder causing patients minor physical discomfort. The prevalence of HSV may be high in innocent infections, as high as 1/3 of the population. However, HSV infection and its complications with troublesome recurrences may make the problem worse. The apparent increase in HSV infection over recent years may be partly due to increased publicity about the disease, the current antiviral treatment, the inclusion of both primary and recurrent cases in clinic follow up and the increased use of viral cultures for diagnosis. The aim of this work is to share in the study of the detection of HSV through virus isolation and detection of antiviral antibodies using IIF technique, as well as the evaluation of the diagnosis by the above mentioned methods. PMID- 16711540 TI - [Concomitant treatment with gemcitabine and radiotherapy in patients with advanced stages of epidermoid carcinoma originating in the head and neck. A phase II sutdy]. AB - BACKGROUND: Surgery, radiotherapy or radiotherapy alone (RTA) constitute conventional treatment regimes for advanced stages of squamous cell carcinoma originating in the head and neck mucosa. Nevertheless, the results in advanced regional carcinoma (ARC) are disappointing. The chemotherapy-radiotherapy (CHT RT) association has shown to be superior to RTA in irresectable disease and in resectable disease it could substitute initial surgery as a rescue alternative. OBJECTIVE: Our objective is to report the response rate and toxicity of concurrent treatment with Gemcitabine and Radiotherapy (GRT) in patients with ARC. In a prospective design, patients with ARC received concurrent GRT; the global, complete and partial response rate as well as toxicity were assessed. MATERIAL AND METHODS: 15 patients were included, 5 women and 10 men, 73% in stage IVa; 13/15 showed a global response (87%), a complete response was observed in 9 cases (60%) (RC) and 2 showed progress. RESULTS: All patients included showed toxicity, the most frequent one was level 4 mucositis in 46%, of this 40% required nutritional support by catheter or gastrostomy. One patient in RC died due to sepsis. None of them suspended treatment. CONCLUSION: The GRT association offers a complete response rate of 60%; nevertheless morbidity was not insignificant; randomized studies with a larger number of patients will be required to allow us to outline the optimal therapeutic scheme. PMID- 16711541 TI - [Assessment of five thrombophilic genetic polymorphisms among couples with habitual abortion]. AB - An association between thrombophilic genes and obstetric conditions with early pregnancy termination has been previously proposed. In the present study we attempted to evaluate the possible association between thrombophilic genetic polymorphisms and habitual abortion (HA). Samples from two groups of volunteers were analyzed. The experimental group (n>100) was conformed by women attending the Centro Medico de Occidente, IMSS and their male couples, with a reproductive history ofat least three miscarriages. The reference group (n > 200) was composed by male and female healthy adults living in the state of Jalisco, Mexico. DNA was extracted from peripheral blood, and polymorphisms FII G20210A , FVG1691A, MTHFR C677T, ECA IID y TNF G-308A were typed by PCR-RFLP or -SSP. Genotype proportions in the reference group were in agreement with the HardyWeinberg expectations. Allele, genotype, and phenotype proportion inter-group comparisons did not show statistically significant differences. The present results could not demonstrate that thrombophilic polymorphisms constitute risk factors for HA in Jalisco. PMID- 16711542 TI - [Treatment of non-Hodgkin's lymphoma associated with acquired immnodeficiency syndrome (AIDS) at the Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran]. AB - BACKGROUND: Survival in patients with acquired immunodeficiency syndrome (AIDS) related non-Hodgkin's Lymphoma has improved with the use of High Active Antiretroviral Therapy (HAART) and less toxic chemotherapy. MATERIAL AND METHODS: Clinical characteristics and outcome among patients treated for AIDS related non Hodgkin's Lymphoma are described. Nine patients were studied retrospectively. Overall survival (OS) and Free Disease Survival (FDS) using a Kaplan-Meier model were analyzed. RESULTS: Patients received (DA-EPOCH) etoposide, prednisone, vincristine, doxorubicin and cyclophosphamide. The overall Survival was 18 months and 13 month Free Disease Survival with a median follow-up of 16 months showing full response in 8/9 patients was observed. CONCLUSIONS: A very satisfactory treatment response in this group of patients expressed as an increased Overall Survival was noted. PMID- 16711543 TI - [Poisoning caused by Latrodectus Mactans (Black Widow) spider bite among children. Clinical features and therapy]. AB - INTRODUCTION: Poisoning caused by "black widow" spider bite, is not commonly observed during childhood, it is not exempt from complications, and must be treated in time or the patient can die. In Mexico we do not have accurate data on the exact number of accidents due to this condition. OBJECTIVE: Identify the signs and characteristic symptoms of children treated at the Children's Hospital from the state of Sonora with the aim of starting early treatment. MATERIAL AND METHODS: We reviewed 43 clinical records of hospitalized patients admitted for "black widow " spider bite between 1978 - 2004. Demographic variables were: age, gender, place of origin, place where accident took place, treatment given prior to hospital admittance, degree of envenomation measured by signs and symptoms, treatment and clinical evolution. RESULTS: 69.7% of cases were male, we did not find differences in the number of patients between infant, pre-school, elementary school children and teenagers. During summer and afternoon hours we registered 44% and 37% cases respectively; 74% patients were poisoned by the spider at home; the anatomical areas most affected were: arms, hands and thorax. Signs and symptoms in infants were: irritability, constanty cry, sialorrea, agitation erythema on wounded areas and seizures. Among elementary school children and teenagers signs included pain in wounded area, abdominal and thoracic, muscle spasms, fine tremor and residual signs. Patients who received fabotherapic treatment had better evolution and less time of hospitalization. No deaths were reported. CONCLUSIONS: The different signs and symptoms between infants, preschool, elementary school children and teenagers allowed us to clearly identify evenommation and differentiate it form confounding diseases. In our study, specific fabotherapy constituted the best treatment choice. PMID- 16711544 TI - [Malingering among workers seeking disability insurance]. AB - OBJECTIVE: Describe the frequency and characteristics of Mexican Social Security workers with malingering disorder that request disability pension. MATERIAL AND METHODS: Comparative survey made among 136 workers seen during 2001, which were divided into three groups: malingering workers (MW), workers without disability (WOD), and workers with disability (WWD). We administered the Z Test for scaled variables and Z2 Test for nominal variables to identify group differences RESULTS: The incidence ofmalingering was of 2.2/100,000 workers. Mean age was 41.9 +/- 10.1 years, 440 was the average number of days of labor disability; 51 (37%) workers were malingerers. 35 (26%) workers were work disabled and 50 (37%) without disability. Malingerers had higher level of schooling compared with WOD and WWD (p < 0.02); most worked at the Social Security (p < 0.05), with lower number of previous jobs (p < 0.05), presented longer work disability (p < 0.05). Depressive symptoms were not frequently noted (p < 0.025) and all had normal electroencephalogram and brain computed tomography studies. CONCLUSIONS: Malingering workers who request work disability pensions follow a particular pattern that differs from other workers that request disability assessment at the Social Security Institute of Mexico. PMID- 16711545 TI - [Impact of the duration of untreated psychosis among patients with a first episode: a one year follow-up study]. AB - BACKGROUND: The delay on the onset of specific treatment has several prognostic implications for psychotic patients. Duration of untreated psychosis (DUP) has been considered a prognostic variable. A longer DUP has been associated with a poor clinical outcome. OBJECTIVE: To determine the DUP in a sample of first episode psychotic patients and its relationship with recovery and relapse on a one-year follow-up study. MATERIAL AND METHODS: A sample of 66 first-episode psychotic patients was recruited and divided according to their diagnoses in affective and non-affective psychoses. Diagnoses were obtained with SCAN and DUP was registered for each patient. A clinical assessment for psychotic and affective symptoms was performed using standardized instruments. RESULTS: All patient studied showed clinical improvement during follow-up. Patients with a longer DUP showed poorer psychosocial adjustment and lower recovery indexes. Treatment suspension was the main variable associated with relapse. CONCLUSIONS: DUP is an important prognostic variable. Early detection programs are required to decrease the period between illness onset, diagnosis and treatment in first episode psychotic patients. PMID- 16711547 TI - [Mexican medical journals]. PMID- 16711546 TI - [Comparative effectiveness of 0.1% metilprednisolone aceponate and 0.05% betamethasone dipropionate among children with nonretractable prepuce]. AB - OBJECTIVE: To compare clinical improvement between treatment with metilprednisolone aceponate vs. betamethasone dipropionate among children with nonretractable prepuce. MATERIAL AND METHODS: Between August 2001 and November 2002, we carried out a double blind and controlled clinical trial in 34 children with a diagnosis of nonretracable prepuce. Children were randomly assigned to one of the following groups and topical treatment was administered: Group A; metilprednisolone aceponate 0. I 1% and Group B; betamethasone dipropionate 0.05%. RESULTS: Improvement was noted in 88.2% of our sample studied; (n= 15) children from group A and 76.4% (n= 13) childrenfrom group B; however, we did not observe a significant difference when comparing percentages between the two groups (chi2 = 0.2; p = 0.6). CONCLUSIONS: The percentage of clinical improvement was similar between the two groups of topical steroid treatment administered. PMID- 16711548 TI - [The role of eotaxins in bronchial asthma and nasal polyposis]. AB - Over the last few years, three specific eosinophil activating peptides, eotaxin 1, -2 and -3, members of the chemokine family have been identified. These cytokines exert a number of functions on eosinophils including chemotaxis, transendothelial migration and induction of the release of reactive oxygen species. Eosinophils are considered to play an important role in allergic disease by causing tissue damage through the release of toxic proteases, lipid mediators, cytokines and oxygen free radicals. This article reviews the role of eotaxins in asthma and nasal polyps. Discussion focuses on therapeutic guidelines, particularly as it has been shown that CCR3, the major chemokine receptor in eosinophils, serves as a eotaxin receptor. PMID- 16711549 TI - [Molecular hemato-oncology and new specific treatment strategies for leukemia]. AB - Leukemia-associated fusion genes are detected in a significant proportion of newly diagnosed cases, where genes encoding transcription factors are usually found at one of the breakpoints. Activated fusion proteins such as Pml-Raralpha have been shown to inhibit cellular differentiation by recruitment of nuclear corepressor complexes, which maintain local histone deacetylase (HDAC) in a variety of hematologic lineage-specific gene promoters. This HDAC-dependent transcriptional repression appears as a common pathway in the development of leukemia and could constitute an important target for new therapeutic agents. Alternatively, the Bcr-Abl oncoprotein shows high tyrosine kinase activity and deregulates signal transduction pathways normally involved in both apoptosis and proliferation. This aberrant activity is affected by signal transduction inhibitors (STIs), which block or prevent the oncogenic pathway. In this review, we shed some light on our understanding of both the reversible transcriptional repression controlled by HDAC and the deregulated Bcr-Abl signal transduction pathway. In addition, the administration of low molecular weight drugs for human leukemia treatment based on this knowledge brings about a significant long-term clinical remission and an acceptable risk of toxic effects that should increase the cure rate. PMID- 16711551 TI - [Thoracic wall metastasis in liposarcoma]. AB - Myxoid liposarcoma often metastasizes to extrapulmonar sites; however, osseous metastases are rare. We report the case of a 52-year-old male with a single histologically proven bone metastasis of the thoracic wall from a primary myxoid liposarcoma with a round cell component of the right thigh. PMID- 16711550 TI - [Nasal cavity angiosarcoma: a case report and literature review]. AB - Sarcomas constitute 5% of all of head and neck neoplasms. Angiosarcomas comprise 2% of all soft tissue tumors, 60% appear in skin and soft tissue, 50% in the head and neck and only 4% are present in the upper aerodigestive tract. We report a case of a 52 year old healthy woman with headache, weeping, foreign body sensation in the left nostril and epixtasis. The TAC identified a tumor in the left nostril that invaded the osseous lamina of the skull. The histopathologic diagnosis was angiosarcoma. We carried out a literature review and analysis from 1976 to 2004. Agiosarcomas of the upper aerodigestive tract have a better prognosis and long term survival free of disease than their soft tissue and skin counterparts. Recurrence is strongly correlated with positive resection margins. Complete surgery and radiotherapy are considered the optimal treatment schemes. PMID- 16711552 TI - [The concept of chronicity and degeneration in neurology]. AB - Chronic degenerative diseases hold a special place in current medicine due to the impact they have in the patient, the familiar and medical environment, and by its social, financial and work related repercussions. Even though medical literature includes numerous scientific writings that approach diverse aspects of these diseases, just a few of them deal with historical or conceptual questions. Following the historical evolution that medical language has undergone will allow us to approach and to study in depth both scientific and medical knowledge. The aim of this study was analyze, from a historical perspective, how two concepts: chronic and degeneration, that have a single origin, are used differently and have a different meaning, nowadays have been combined to describe diseases with an underlying biological process, generally known as degenerative, and which in addition share a temporal condition. In this sense chronic degenerative diseases offer medicine new challenges and different ways to approach them. PMID- 16711553 TI - [Oral cholangiography and duodenal atresia]. AB - A newborn male patient with trisomy-21 presented with bilious hemesis. The patient was icteric with slight hepatomegaly. Simple abdominal X-ray and upper gastrointestinal series with barium showed a dilated duodenal loop and inflammatory changes involving the duodenal mucosa. This image known as "double bubble" is characteristic of congenital duodenal obstruction. Simultaneously the gallbladder and choledochus were visualized. The former X-ray finding is very unusual. An uneventful Kimura procedure was performed. PMID- 16711554 TI - [Tamoxifen and endometrial cancer]. PMID- 16711555 TI - The demise of "surgical dentistry". PMID- 16711556 TI - Denis/SADA dilemma. PMID- 16711557 TI - Dietary intakes and caries experience in children in Limpopo Province, South Africa. AB - INTRODUCTION: The identification of a rural village with a very low incidence of dental caries among the children provided the opportunity to study the diets of children apparently not exposed to risk factors for dental caries and compare them with children who were at risk. OBJECTIVE: To compare the diet, nutritional status and dental caries experience of 10- and 15-year-olds in an urban town (Malamulele) and a rural village (Mahonisi) in the Limpopo Province, South Africa. METHODS: Dental examinations and anthropometric measurements were done according to standard procedures. Dietary intakes were assessed by four 24-hour recalls. RESULTS: Mean dft/DMFT (decayed and filled deciduous teeth/Decayed, missing and filled permanent teeth) scores were 1.21 and 0.65 for 10- and 15-year olds in urban Malamulele compared to 0.33 and 0.02 in rural Mahonisi. The percentage caries free in Malamulele was 36% compared to 88% in Mahonisi. The children in Mahonisi were shorter and lighter than those in Malamulele. Estimated fluoride intakes were significantly lower in Malamulele than in Mahonisi (p = 0.01). Mean total added sugar (all sugar not naturally occurring in foods) intake in Malamulele was significantly higher than in Mahonisi (p < 0.05). CONCLUSION: The most significant dietary differences between the two areas were total added sugar and fluoride intakes. The lower added sugar and higher fluoride intake among the rural Mahonisi children contributed to the lower dental caries experience. PMID- 16711558 TI - Posterior composite restorations and post-operative sensitivity. PMID- 16711559 TI - Fordyce granules on the prolabial and oral mucous membranes of a selected population. AB - Fordyce granules are commonly seen on the prolabial and oral mucous membranes of adults. According to Sewerin there is a prevalence of 80% on the prolabial mucous membranes and 95% on the oral mucous membranes and reaches a peak between 20 and 29 years of age. Experienced clinicians find that these figures are too high for the South African population. In order to accept or reject this hypothesis, it was decided to determine the prevalence of Fordyce granules in a selected population of adult South-Africans of all the age groups, but mainly between 20 and 29 years of age. In this study, the prevalence of ectopic sebaceous glands (Fordyce granules) on the prolabial and oral mucous membranes of a selected South African population, was 62.07% and 57.25% in adult males and females, respectively. This is significantly lower than the prevalence reported by Sewerin. This study also revealed that Fordyce granules are mostly seen on the lips (vermillion), labial mucosa and the corresponding parts of the alveolabial sulcus. The buccal mucosa is the second most common site, and there is no significant difference in the prevalence between males and females. PMID- 16711560 TI - Implant removal. PMID- 16711562 TI - General Practitioners radiology. Case 39. Diagnosis. Palatal torus. PMID- 16711561 TI - General practitioner's pathology. Case 2. Diagnosis: Oral Kaposi's sarcoma with secondary ulceration. PMID- 16711563 TI - Sedation corner 7. Apisectomy. PMID- 16711564 TI - Brief motivational interviewing: motivating behaviour change. PMID- 16711565 TI - [Risk factors for clinical signs of PMWS and PDNS in pigs in The Netherlands: a case-control study]. AB - Potential risk factors for clinical signs of post-weaning multisystemic wasting syndrome (PMWS) and porcine dermatitis and nephropathy syndrome (PDNS) in pigs in the Netherlands were investigated in a matched case-control study using a questionnaire (personal interview). Eighty-two pig farmers were questioned about management, hygiene, husbandry systems, disease history, and preventive health care. In this study, 30 pig herds with (cases) and 30 pig herds without (controls) characteristic clinical signs of PMWS were compared. For PDNS, 11 pig herds with (cases) and II pig herds without (controls) characteristic clinical signs of PDNS were compared. Univariate analysis (P < 0.10) showed that the following occurred relatively more often in the PMWS case herds than in the control herds: 1) clinical signs of PDNS, porcine reproductive and respiratory syndrome (PRRS), porcine parvovirus (PPV) infections, meningitis, coccidiosis, and pre-weaning diarrhoea observed by the farmer; 2) vaccination against PRRS and mycoplasma; 3) non-optimal climatic conditions in the nursery rooms, a large variation in weaning age, a high occurrence of cross-fostering of piglets, a large number of sows with lactation problems, poor colostrum intake by piglets; and 4) (historical) use of breeding stock (including semen for artificial insemination) of Anglo-Saxon origin. In the final multivariate statistical model, one variable remained significantly associated with PMWS case herds, namely, the presence of clinical signs of PRRS (and/or the associated use of vaccination against PRRS). It should be noted that in almost all cases animals were vaccinated against PRRS because of clinical signs of PRRS that appeared a few months after the first occurrence of clinical signs of PMWS. This excludes PRRS vaccination as a primary factor in causing PMWS. Analysis of the PDNS case control data showed comparable results with those of the PMWS study. In the final statistical model, the presence of clinical signs of PRRS (and/or the associated use of vaccination against PRRS) was significantly associated with PDNS case herds. PMID- 16711567 TI - [Visibly blind?]. PMID- 16711566 TI - [Mental notes in tracking BVDV carriers]. AB - The detection and elimination of animals persistently infected with bovine viral diarrhoea virus (BVDV) is the key to BVD control. This method has proven to be very efficient in eradicating BVDV in a herd. Several pitfalls in the detection procedure can make that some persistently infected (PI) animals do not get identified or are removed too late, supporting the assumption that circulation of the virus could be possible in absence of PI animals. Furthermore the risk of reintroduction is high since the prevalence of BVD is high in the Netherlands and Belgium. Based on both practical experience and literature, here we review critical control points in order to minimise the risk of a false negative BVDV screening. PMID- 16711568 TI - [Emergency euthanasia and group practice veterinarians]. PMID- 16711569 TI - [Less distance between veterinary communicable diseases 'science' and 'practice' very important. Interview by Marianne M. Sloet van Oldruitenborgh-Oosterbaan]. PMID- 16711570 TI - [Threatening classical swine fever from Germany. Congressman Veerman makes PCR test for classical swine fever possible]. PMID- 16711571 TI - [Ritual slaughter]. PMID- 16711572 TI - [Star clinics AUV]. PMID- 16711573 TI - DoH draws a blank in quest to attract outsiders to SHA posts. PMID- 16711574 TI - Trusts face clinical data penalties. PMID- 16711576 TI - Politics. Why the NHS is on the ballot paper. PMID- 16711575 TI - Debt-hit counties plan trauma cuts. PMID- 16711577 TI - On the last of the great PFIs. PMID- 16711578 TI - Barometer. PCTs April 2006. PMID- 16711579 TI - Primary care. How to find 'frequent fliers'. AB - So-called 'frequent fliers' account for a Large proportion of unplanned admissions. Work in Manchester suggests age is not the best indicator for case management--social deprivation and mental illness may be more useful. PMID- 16711580 TI - On primary care. PMID- 16711581 TI - Diversity. How to break a glass ceiling. PMID- 16711582 TI - Clinical leadership. Structural instability. PMID- 16711584 TI - Medicaid funding may be non-issue this year. PMID- 16711583 TI - Going the extra mile for patients with special needs. PMID- 16711585 TI - DEC mandates use of amalgam separators. PMID- 16711586 TI - Laser treatment of atypical oral pigmentation. A case report. AB - A 46-year-old female Caucasian patient presented with severe black pigmentation of the maxillary attached gingiva. The discoloration extended from tooth #2 to tooth #15 and was noted only on the buccal aspect of the arch. The patient reported that her gingival tissues appeared normal until age 22, at which time her first child was born. Cosmetic considerations prompted by this child's impending marriage caused the patient to seek treatment. It was important to determine the cause of the pigmentation so that we would be reasonably assured that it would not recur. A CO2 laser was used to remove the pigmented tissue in such a manner that the postoperative period was both painless and rapid. PMID- 16711587 TI - Task force on special dentistry makes meaningful improvement. AB - This article explores the formation of a task force to address the specific issues that confront dentists who treat patients with disabilities. From the first meeting, this all-volunteer task force, with representatives from clinical dentistry, public health groups, hospitals and dental schools, set out to recognize the specific issues that arise with special needs patients and offer workable solutions. This article enumerates these issues, and can serve as a reference for dentists and other health professionals. PMID- 16711588 TI - New York State Office of Mental Retardation and Developmental Disabilities. A brief history and timeline. PMID- 16711589 TI - Providing dental care to patients with developmental disabilities. An introduction for the private practitioner. AB - During the past 20 to 25 years, in New York State, most individuals with developmental disabilities who had previously been housed in large institutions have been mainstreamed into community-based residences. This shift has created a need for dental services in the community. The article presented here provides information and tips to assist the general practitioner with integrating these patients into a private practice setting. PMID- 16711590 TI - Dental considerations for individuals with Down syndrome. AB - Down syndrome is one of the most frequently encountered and easily recognizable of all developmental disabilities. Patients with Down syndrome exhibit mental retardation and often present with associated medical conditions, such as cardiac defects, immune deficiencies and musculoskeletal disorders. It is important for the dental practitioner to be familiar with the medical aspects of Down syndrome, as well as the potential behavioral issues. We present a brief review of Down syndrome for the dental provider and suggest that with a little knowledge and some patience, most private practitioners can easily accommodate these patients in their practice. PMID- 16711591 TI - Providing dental care to patients with developmental disabilities: medical/legal issues. AB - Dentists providing treatment to individuals with developmental disabilities are often faced with unique medical/legal issues. Obtaining informed consent when a patient does not have capacity can be an involved process. Issues regarding therapeutic aids used for immobilization during treatment may further complicate the situation. PMID- 16711592 TI - Effect of preventive dentistry training program for caregivers in community facilities on caregiver and client behavior and client oral hygiene. AB - The influx of persons with developmental disabilities into community-based programs has required the development, expansion and improved integration of community-based services. The role of caregivers in these community settings has become pivotal. However, with regard to daily activities, oral hygiene is often a low priority. Few attempts have been made to study caregiver participation in oral disease prevention and practices. This study evaluated the effects of an indirect training program, where caregivers were trained but the ultimate effects were demonstrated in adult clients. In addition, these effects were demonstrated in community care settings. A multiple baseline design, across three group homes, included 11 adult clients with developmental disabilities. This study evaluated the effects of training, instruction to use training and coaching on the presence of caregivers during oral hygiene sessions, the duration of toothbrushing and plaque scores of the clients. The results of this investigation demonstrated that there was an increase in caregiver presence and duration of brushing with a concurrent decrease in plaque scores. Caregivers responding to social validation questionnaires believed that 8 out of 11 clients had increased skills. This study demonstrates that caregiver training, combined with specific instructions to use training information and coaching of caregivers, can have a positive impact on the oral health of individuals with developmental disabilities living in community settings. PMID- 16711594 TI - In service to others. PMID- 16711593 TI - Slipping through the cracks. Dental care for older persons with intellectual disabilities. AB - An increasing population of men and women with intellectual disabilities and other developmental disabilities is reaching older ages. As our country continues the process of deinstitutionalization, these individuals will require treatment in communities. Dental practitioners increasingly will be involved in the needs of this population, many of whom are members of families being treated in private dental practices. A review of these developments and the particular dental needs of these patients is considered. PMID- 16711596 TI - Effects of geographic source, rearing system, and season on the nutritional quality of wild and farmed Perca fluviatilis. AB - The effects of season, geographic source (Lake Geneva, Rhine River), and rearing system (extensive, semiextensive, and intensive systems) on the lipid content and FA composition of fillets of Perca fluviatilis were studied. Significant differences in the total lipid content were found between fish coming from the Rhine River and Lake Geneva (1.21 and 1.48%, respectively). Seasonal effects were investigated quarterly for perch sampled in the Rhine River. Intensively reared perch displayed a higher lipid content (1.48%) than the other farmed perch, i.e., 1.26% for a semiextensive system and 1.16% for an extensive system. No significant difference in lipid content was found (i) between lacustrine fish and intensively reared fish or (ii) among fish from the Rhine River and the semiextensive or extensive rearing systems. The main FA were 22:6n-3 (DHA, 21.3 37.1% of total FA), 16:0 (17.7-20.2%), 20:5n-3 (EPA, 9.2-13.2%), 18:1 (8.0 11.5%), 20:4n-6 [arachidonic acid (ARA), 1.9-10.7%], 16:1 (4.3-6.0%), and 18:2n-6 (2.1-6.0%). In comparison with perch coming from the Rhine River, the lacustrine fish were characterized by higher total n-6 PUFA and a lower proportion of both total monounsaturated FA (MUFA) and total n-3 PUFA. Among rearing systems, extensively farmed fish had higher n-6 PUFA and lower n-3 PUFA contents. Wild fish showed higher ARA and 18:2n-6 than farmed fish. They also had significantly more EPA (12.5-13.2%) than farmed perch (9.2-10.9%). For DHA, no difference existed between (i) the lacustrine fish (31.9% of total FA) and the intensively reared fish (33.0%) and (ii) the Rhine (37.1%) and semiextensively reared fish (36%). Effects of size, diet composition, and environmental conditions on the total lipid contents and FA composition are discussed. PMID- 16711595 TI - Vitamin A deficiency enhances docosahexaenoic and osbond acids in liver of rats fed an alpha linolenic acid-adequate diet. AB - The synthesis of docosahexaenoic (DHA, 22:6n-3) and Osbond acid (OA, 22:5n-6) is regulated by the heterodimer of peroxisome proliferator-activated receptor and retinoid X receptor (RXR). 9-Cis retinoic acid, a metabolite of vitamin A, is the most potent ligand of RXR. We tested whether vitamin A deficiency impairs DHA and OA synthesis in rats fed a vitamin A- and alpha-linolenic acid (ALA)-sufficient (VASALAS), vitamin A-sufficient and ALA-deficient (VASALAD), vitamin A-deficient and ALA-sufficient (VADALAS), or vitamin A- and ALA-deficient (VADALAD) diet. After 7 wk of feeding, liver and colon choline (CPG) and ethanolamine (EPG) phosphoglyceride FA were analyzed. The VADALAS compared with the VASALAS rats had elevated levels of both DHA (P< 0.05) and OA (P < 0.005) in liver CPG and EPG. In contrast, the VADALAD group had a lower DHA (P < 0.01) and higher OA (P < 0.005) level in CPG and EPG of both tissues than their VASALAD counterparts. ALA deficiency reduced DHA and enhariced OA levels in liver and colon CPG and EPG in both the vitamin A-sufficient (VASALAS vs. VASALAD) and -deficient (VADALAS vs. VADALAD) rats (P < 0.005). The study demonstrates that ALA deficiency reduced DHA and enhanced OA levels in tissue membranes, and dietary vitamin A deficiency has a profound effect on membrane DHA and OA in rat tissues. Both vitamin A and DHA are involved in a myriad of vital physiological functions pertaining to growth and development and health. Hence, there is a need for a further study to unravel the mechanism by which vitamin A influences membrane DHA and OA. PMID- 16711597 TI - Fatty acid profiles and relative mobilization during fasting in adipose tissue depots of the American marten (Martes americana). AB - The American marten (Martes americana) is a boreal forest marten with low body adiposity but high metabolic rate. The study describes the FA composition in white adipose tissue depots of the species and the influence of food deprivation on them. American marten (n = 8) were fasted for 2 d with 7 control animals. Fasting resulted in a 13.4% weight loss, while the relative fat mass was >25% lower in the fasted animals. The FA composition of the fat depots of the trunk was quite similar to other previously studied mustelids with 14:0, 16:0, 18:0, 16:1 n-7, 18:1 n-9, and 18:2n-6 as the most abundant FA. In the extremities, there were higher proportions of monounsaturated FA (MUFA) and PUFA. Food deprivation decreased the proportions of 16:0 and 16:1 n-7, while the proportion of long-chain MUFA increased in the trunk. The mobilization of FA was selective, as 16:1 n-7, 18:1 n-9, and particular n-3 PUFA were preferentially mobilized. Relative mobilization correlated negatively with the carbon chain length in saturated FA (SFA) and n-9 MUFA. The delta9-desaturation of SFA enhanced the mobilization of the corresponding MUFA, but the positional isomerism of the first double bond did not correlate consistently with relative mobilization in MUFA or PUFA. In the marten, the FA composition of the extremities was highly resistant to fasting, and the tail tip and the paws contained more long-chain PUFA to prevent the solidification of lipids and to maintain cell membrane fluidity during cooling. PMID- 16711599 TI - Olive oil consumption and weight change: the SUN prospective cohort study. AB - The aim of this dynamic prospective follow-up study was to assess the association between olive oil consumption and the likelihood of weight gain or the incidence of overweight or obesity in a large Mediterranean cohort of 7,368 male and female Spanish university graduates (the SUN Project) who were followed for a median period of 28.5 mon. A validated Food Frequency Questionnaire was administered at baseline, and respondents also completed a follow-up questionnaire after 28.5 mon. Changes in participants' consumption of olive oil and their weight were assessed during follow-up. A higher baseline consumption of olive oil was associated with a lower likelihood of weight gain, although the differences were not statistically significant. The adjusted difference in weight gain (kg) was 0.16 [95% confidence interval (CI): -0.42 to +0.11] for participants in the upper quintile of olive oil consumption (median: 46 g/d) compared with those in the lowest quintile (median: 6 g/d). For participants with a high baseline consumption of olive oil whose olive oil consumption also increased during follow up, we found a slightly increased but nonsignificant risk of incidence of overweight or obesity (adjusted odds ratio = 1.19, 95% CI: 0.73 to 1.95). Our study, carried out in a sample of free-living people, shows that a high amount of olive oil consumption is not associated with higher weight gain or a significantly higher risk of developing overweight or obesity in the context of the Mediterranean food pattern. PMID- 16711598 TI - Effects of conjugated linoleic acid and troglitazone on lipid accumulation and composition in lean and Zucker diabetic fatty (fa/fa) rats. AB - Dietary CLA has been shown to enhance glucose tolerance in several animal models, but in mice it induces insulin resistance and lipodystrophy. In this study, the effects of 2 wk of diet supplementation with either 1.5% CLA or 0.2% troglitazone (TZD), an insulin-sensitizing thiazolidinedione, on glucose tolerance, lipid accumulation, and composition of both lean and Zucker diabetic fatty (fa/fa; ZDF) rats were examined. Compared with lean rats, which maintained normal glucose tolerances after 2 wk of feeding regardless of diet, ZDF rats fed a control diet (CON) had significantly worsened glucose tolerance. ZDF rats fed CLA and TZD diets, however, maintained normal glucose tolerances. In contrast to the significantly elevated lipid levels in ZDF rats fed the CON diet, concentrations of plasma FFA and TG in ZDF rats fed CLA and TZD diets were normalized. A similar reduction of plasma lipid levels was observed in lean rats fed CLA and TZD compared with lean rats fed the CON diet. Although ZDF CON rats developed significant hepatic steatosis, both CLA- and TZD-fed rats had hepatic TG levels similar to those of lean rats. Both lean and ZDF rats fed the CLA diet had reduced adipose mass compared with respective genotype controls; however, TZD had no effect. Ratios of 16:1/16:0 and 18:1/18:0 FA, surrogate markers for stearoyl CoA desaturase-1 (SCD-1) activity, were reduced in livers of ZDF rats fed CLA and TZD diets. These results show that, like TZD, CLA normalizes glucose tolerance and plasma lipids and also improves hepatic steatosis and FA composition in ZDF rats. The effects of CLA and TZD on hepatic lipid composition suggest that the effects of these two agents on glucose tolerance may be associated with a reduction in SCD-1. PMID- 16711601 TI - Oleate-induced formation of fat cells with impaired insulin sensitivity. AB - Exogenous FA cause lipid accumulation in pre-adipocytes. We investigated whether the fat cells thus formed are metabolically distinct from adipocytes differentiated with standard methylisobutylxanthine, dexamethasone, and insulin (MDI) hormonal cocktail by comparing their expression of adipogenic genes, accumulation of TAG, lipogenesis, lipolysis, glucose uptake, and the effects of insulin on selected metabolic activities. Cells exposed to oleate began to accumulate TAG in parallel or prior to the induction of adipogenic genes, whereas cells treated with MDI expressed adipogenic genes before TAG accumulation. Oleate treated fat cells also showed exaggerated basal lipolysis and weak response to insulin in both lipolysis regulation and glucose uptake. These findings were associated with increased basal phosphorylation of perilipin, increased Glut-1 but decreased Glut-4 expression, and reduced insulin-induced Akt phosphorylation. We suggest that this unique fat cell phenotype might be a mimetic of what can happen to fat cells formed in vivo under the influence of circulating FA and might be a useful model for in vitro studies of obesity-related insulin resistance in adipocytes. PMID- 16711600 TI - Effects of soy or milk protein during a high-fat feeding challenge on oxidative stress, inflammation, and lipids in healthy men. AB - Soy isoflavones may impede atherogenic processes associated with cardiovascular disease. Research suggests that the postprandial generation of TG-rich remnants contributes to the development of atherosclerosis. The purpose of the current study was to determine if 39 g soy (85 mg aglycone isoflavones, treatment) compared with 40 g milk protein (0 mg aglycone isoflavones, control) in combination with a high-fat meal can modify postprandial, atherogenic-associated events and biomarkers for oxidative stress, inflammation, and thrombosis. Fifteen healthy men (20-47 yr) participated in a double-blind cross-over meal-challenge study occurring on two nonconsecutive days. The study meals consisted of two high fat apple muffins consumed with either a soy or milk shake (229 mL, 41% fat, 41% carbohydrate, and 18% protein). Blood samples were obtained at baseline (fasted) and hours two, four, and six postprandial. Plasma TG significantly increased in both treatment and control meal challenges compared with baseline. There were no significant differences (P > 0.05) between treatment (soy) and control (milk) for ex vivo copper-induced LDL oxidation, serum C-reactive protein, serum interleukin 6 (IL-6), serum fibrinogen, or plasma lipids (total cholesterol, HDL, LDL, TG). IL-6-concentrations significantly decreased as a function of time during either meal challenge (P = 0.005). These data suggest that consumption of soy or milk protein in conjunction with a high-fat meal does not acutely modify postprandial oxidative stress, inflammation, or plasma lipid concentrations in young, healthy men. PMID- 16711602 TI - Coexistence of phosphatidylcholine-specific phospholipase C and phospholipase D activities in rat cerebral cortex synaptosomes. AB - DAG derived from phosphatidylcholine (PtdCho) acts as a lipid second messenger. It can be generated by the activation of phospholipase D (PLD) and the phosphatidic acid phosphohydrolase type 2 (PAP2) pathway or by a PtdCho-specific phospholipase C (PtdCho-PLC). Our purpose was to study PtdCho-PLC activity in rat cerebral cortex synaptosomes (CC Syn). DAG production was highly stimulated by detergents such as Triton X-100 and sodium deoxycholate. Ethanol and tricyclodecan-9-yl-xanthate potassium salt decreased DAG generation by 42 and 61%, respectively, at 20 min of incubation. These data demonstrate that both the PLD/PAP2 pathway and PtdCho-PLC contribute to DAG generation in CC Syn. PtdCho PLC activity remained located mainly in the synaptosomal plasma membrane fraction. Kinetic studies showed Km and Vmax values of 350 microM and 3.7 nmol DAG x (mg protein x h)(-1), respectively. Western blot analysis with anti-PtdCho PLC antibody showed a band of 66 KDa in CC Syn. Our results indicate the presence of a novel DAG-generating pathway in CC Syn in addition to the known PLD/PAP2 pathway. PMID- 16711603 TI - Diacylglycerol acyltransferase activity and triacylglycerol synthesis in germinating castor seed cotyledons. AB - The central importance of storage lipid breakdown in providing carbon and energy during seed germination has been demonstrated by isolating the genes encoding the enzymes involved in FA beta-oxidation. In contrast, little is known about the ability of germinating seeds to synthesize TAG. We report that castor cotyledons are capable of TAG synthesis. The rate of incorporation of ricinoleic acid into TAG reached a peak at 7 d after imbibition (DAI) (1.14 nmol/h/mg) and decreased rapidly thereafter, but was sustained at 20 DAI in cotyledons and true leaves. The castor DAG acyltransferase (RcDGAT) mRNA and protein were expressed throughout seed germination at levels considerably enhanced from that in the dormant seed, thus indicating new expression. Significant degradation of the RcDGAT protein was observed after 7 DAI. The DGAT activity was found to be predominantly a function of the level of the intact RcDGAT protein, with the rate of TAG synthesis decreasing as degradation of the RcDGAT protein proceeded. A possible mechanism for the degradation of the RcDGAT protein is discussed. The induction of DGAT mRNA and protein, the capacity for TAG synthesis in vitro and in tissue slices, and the differing TAG composition of dormant seed TAG vs. cotyledonary TAG provide strong circumstantial evidence for active TAG synthesis by cotyledons. However, we have not yet determined the physiological significance of this capability. PMID- 16711604 TI - Prediction of adipose tissue composition using Raman spectroscopy: average properties and individual fatty acids. AB - Raman spectroscopy has been used for the first time to predict the FA composition of unextracted adipose tissue of pork, beef, lamb, and chicken. It was found that the bulk unsaturation parameters could be predicted successfully [R2 = 0.97, root mean square error of prediction (RMSEP) = 4.6% of 4 sigma], with cis unsaturation, which accounted for the majority of the unsaturation, giving similar correlations. The combined abundance of all measured PUFA (> or = 2 double bonds per chain) was also well predicted with R2 = 0.97 and RMSEP = 4.0% of 4 sigma. Trans unsaturation was not as well modeled (R2 = 0.52, RMSEP = 18% of 4 sigma); this reduced prediction ability can be attributed to the low levels of trans FA found in adipose tissue (0.035 times the cis unsaturation level). For the individual FA, the average partial least squares (PLS) regression coefficient of the 18 most abundant FA (relative abundances ranging from 0.1 to 38.6% of the total FA content) was R2 = 0.73; the average RMSEP = 11.9% of 4 sigma. Regression coefficients and prediction errors for the five most abundant FA were all better than the average value (in some cases as low as RMSEP = 4.7% of 4 sigma). Cross correlation between the abundances of the minor FA and more abundant acids could be determined by principal component analysis methods, and the resulting groups of correlated compounds were also well-predicted using PLS. The accuracy of the prediction of individual FA was at least as good as other spectroscopic methods, and the extremely straightforward sampling method meant that very rapid analysis of samples at ambient temperature was easily achieved. This work shows that Raman profiling of hundreds of samples per day is easily achievable with an automated sampling system. PMID- 16711605 TI - Fluorescent image analysis of lipid hydroperoxides in fish muscle with 3-perylene diphenylphosphine. AB - A fluorescent image analysis method was developed to evaluate lipid hydroperoxide formation in fish muscle. The lipid hydroperoxides generated in white and dark fish muscles during storage at 5-6 degrees C oxidized 3-perylene diphenylphosphine located in the tissue to yield the fluorescent derivative, 3 perylene diphenylphosphine oxide (3-PeDPPO). 3-PeDPPO thus obtained was determined by digital fluorescent image analysis. The 3-PeDPPO fluorescence intensity of white and dark muscle increased during low-temperature storage (0-24 h) and was clearly correlated with total lipid hydroperoxide levels in muscle extracts, which were determined by using HPLC based on a triphenylphosphine oxidation method (R2 = 0.954). These results suggest that 3-PeDPPO fluorescence, coupled with fluorescent image analysis, is a novel tool for direct determination of lipid hydroperoxides in fish muscle without a need for extraction of lipid. PMID- 16711606 TI - Acyl migration during debenzylation of 1,3-di-O-benzyl-2-O-acylglycerols. AB - In investigating the preparation of 2-acylglycerol by debenzylation of 1,3-di-O benzyl-2-O-acylglycerol, twelve 1,3-di-O-benzyl-2-O-acylglycerols were synthesized. 2-O-Acyl moieties with one or more protons at the carbon atom adjacent to carbonyl group were found to migrate from C-2 to C-1 of the glycerol moiety during hydrogenation under Pd/C at room temperature. PMID- 16711607 TI - Very long chain fatty acid methyl esters in transesterified palm oil. AB - Some unidentified minor compounds have been observed in the residue from short path distillation of transesterified palm oil that are not detected in the original palm oil. A method combining short-path distillation to enrich the unknowns with fractionation using solid-phase extraction is described. The fractionated components were identified using GC coupled with MS. The transesterified palm oil was found to contain methyl esters of up to C32 carbon atoms. In the very long chain FAME with carbon numbers > or = 20, both even and odd carbon numbers accounted for 0.26 wt%, with C24 and C26 being the major ones present in the residue after short-path distillation of transesterified palm oil. PMID- 16711608 TI - Radiographic techniques of the nasal cavity and sinuses. AB - Diseases of the nasal cavity and sinuses can be frustrating problems for owners and veterinarians alike. Initial physical examination and routine blood work are often unrewarding in directing the clinician at a diagnosis. Diagnostic imaging can help characterize the disease process and direct treatment or further diagnostics. Radiographs of the skull are an excellent first-line diagnostic step in the analysis of nasal disease. The following article reviews basic techniques, positioning, and interpretation of radiographs of the nasal cavity and paranasal sinuses. PMID- 16711609 TI - Nasal computed tomography. AB - Chronic nasal disease is often a challenge to diagnose. Computed tomography greatly enhances the ability to diagnose chronic nasal disease in dogs and cats. Nasal computed tomography provides detailed information regarding the extent of disease, accurate discrimination of neoplastic versus nonneoplastic diseases, and identification of areas of the nose to examine rhinoscopically and suspicious regions to target for biopsy. PMID- 16711610 TI - Basics in canine and feline rhinoscopy. AB - Patients suffering from upper respiratory disease such as chronic nasal congestion, sneezing, nasal discharge, and epistaxis invite complete evaluation of their paired nasal cavities. Thorough assessment of these cavities employs sundry diagnostic procedures that enable the investigating clinician to characterize the internal structures of the nasal cavities. After the conscious patient undergoes a complete physical examination, a gross assessment of its external nasal structures is established and areas of physical asymmetry are noted. A working anatomic knowledge of these asymmetric foci helps to guide the next diagnostic steps. The patient is then placed under general anesthesia, during which, in list order, imaging studies, rhinoscopy, and nasal biopsy or foreign body retrieval, are performed. PMID- 16711611 TI - Surgical considerations of nasal discharge. AB - Surgery of the nasal cavity may be exposed primarily via a dorsal or ventral approach. Surgical treatment of lesions of the nasal cavity usually is limited to benign lesions or in combination with adjunctive therapy such as radiation therapy. Caution must be exercised with a dorsal approach to the nasal cavity to avoid complications of inadvertent penetration of the brain case. Gentle tissue handling and careful closure of the mucoperiosteum must be exercised following a ventral approach to minimize the risk of oronasal fistula formation. PMID- 16711612 TI - Chronic rhinitis in cats. AB - Chronic rhinitis is a common and important problem in cats potentially resulting from a number of intranasal or systemic disorders. Idiopathic chronic rhinosinusitis and nasal neoplasia are the most common causes of chronic nasal disease in cats. For most cats with chronic rhinitis, diagnostic imaging, endoscopic studies, and nasal biopsy will be required to establish a diagnosis. A discussion of some of the more common causes of chronic nasal disease is presented. PMID- 16711613 TI - Canine chronic inflammatory rhinitis. AB - Chronic inflammatory rhinitis is commonly found in dogs with chronic nasal disease and is characterized by lymphoplasmacytic infiltrates in the nasal mucosa in the absence of an obvious etiologic process. The pathogenesis of lymphoplasmacytic rhinitis remains unknown. Animals respond poorly to antibiotics, oral glucocorticoids, and antihistamines, making primary infectious, immune-mediated, or allergic etiologies unlikely. Aberrant immune response to inhaled organisms or allergens may induce inflammation in some animals. Common clinical signs include nasal discharge, sneezing, coughing, epistaxis, and stertor. Diagnosis is made by performing a thorough history, physical examination, radiography or advanced imaging (via computed tomography or magnetic resonance imaging), rhinoscopy, and nasal mucosal biopsy to rule out primary etiologies of nasal discharge. Treatment strategies have included various antibiotics, antihistamines, oral and inhalant steroids, nonsteroidal antiinflammatories, and antifungal medications. Some dogs may respond partially to doxycycline or azithromycin, although it is unclear whether response is related to antimicrobial or antiinflammatory properties of these drugs. Hydration of the nasal cavity through nasal drops or aerosols may limit nasal discharge, and some animals may improve with inhalant (but rarely oral) glucocorticoids. PMID- 16711614 TI - Canine nasal aspergillosis. AB - Chronic nasal discharge is a common clinical sign of disease in dogs. Canine sinonasal aspergillosis is a relatively common disease in dogs. The three hallmarks of canine nasal aspergillosis are a profuse mucoid to hemorrhagic chronic nasal discharge that may alternate with periods of epistaxis, ulceration of the external nares with crusting, and pain or discomfort in the facial region. Diagnostic imaging (preferably computed tomography, CT) of the nasal cavity and paranasal sinuses is an important component of the evaluation of dogs with signs of nasal disease. Rhinoscopy is an important part of both the diagnosis and the therapy for nasal aspergillosis. Therapeutic recommendations for sinonasal aspergillosis have included surgery and the use of several systemic and topical antifungal drugs. PMID- 16711615 TI - Canine and feline nasal neoplasia. AB - Dogs and cats of our society have outgrown their status as merely pets and are now considered our close companions and even family members. This shift in their roles has led to pet owners seeking improved preventative medicine for their four legged friends. Subsequently, dogs and cats are living longer lives than ever before and developing more old-age-related diseases. One of the most devastating diseases of older animals is cancer. Once a veterinarian has detected cancer in a pet, pet owners seek advice on their next course of action. This article is intended to provide concise information regarding the diagnosis and treatment of intranasal tumors of the dog and cat. This article outlines the forms of nasal tumors that are the most common, the recommended imaging and biopsy techniques to diagnose the tumor, and the most appropriate treatments of them. PMID- 16711616 TI - Linear parietal skull fracture in a three-month-old without a history of injury- abuse or not? The issue of a corroborated history. AB - When the clinician is faced with an infant with a skull fracture and there is no history of injury, or the history is provided only by a caretaker, the available data would indicate that a simple linear parietal skull fracture without any accompanying neurologic injury is possible from a fall of at least three feet onto a hard surface. Any more serious, multiple or complex injuries should be treated as suspect and investigated by social services and law enforcement. The clinician should refrain from making any dogmatic statements about what may or may not have occurred. We simply do not have the data to say anything further. It is up to social services and the legal system to investigate and decide if child abuse has occurred. PMID- 16711617 TI - Assisted reproductive technology (ART) in the upstate: reducing the risks of multiple births. AB - Multiple pregnancies from ART procedures are all too common. Efforts to reduce high-order multiple pregnancies will require both social and medical reforms. In lieu of a legislative agenda to reduce multiple pregnancies in the United States, efforts must come from the personnel within the individual ART clinics. The approach to the problem includes accepting a lower pregnancy rate than otherwise might be obtainable, if the observed rate of high-order multiple pregnancies is above a certain threshold. Expanding the opportunities for ART reimbursement will ultimately address this problem in the United States, but until then, it is a problem that will not go away without commitment and resolve. PMID- 16711618 TI - How to guide patients for online information: focus on chronic disease. AB - The purpose of this study was to evaluate the accessibility and utility of a common Internet search strategy so physicians might provide this guidance to their patients seeking health information on chronic disease. We selected three common chronic diseases: diabetes, hypertension, and osteoarthritis. We then formulated basic questions from a patient perspective about diagnosis, treatment, and prognosis. A non-medical professional attempted to answer these questions by typing the disease name into the Google search engine. Focusing on the first ten web sites on the Google list was a successful strategy for finding sites that could answer the study questions (average of 61% of the time). The average number of clicks required to answer any question was 1.6. Web sites sponsored by government agencies (89% success) and hospitals (100% success) yielded answers to the questions more often than other sites. In conclusion, despite the multitude of web sites that appear when the name of a chronic disease is entered into a search engine, the top ten results are usually adequate to provide relevant information on common clinical questions to the average patient. Physicians can guide patients to relevant information by instructing them to type the name of the disease itself into a search engine and focusing on government and hospital sponsored Web sites. PMID- 16711619 TI - Computers and medical coding G codes are coming (from CMS). PMID- 16711620 TI - Cardiology standards. PMID- 16711621 TI - Polydoreiphilia: a disease affecting health care financing. AB - In our country, thinking on the subject of health care financing is confused to the point of constituting a disease, termed polydoreiphilia. Though the disease has multiple manifestations, it is caused by one underlying abnormal mechanism: the irrational love of multiple free gifts. The name of the disease is derived from that mechanism. Practical, realistic, non-ideological measures may help manage the disorder and lead to provision of effective health care for our citizens. PMID- 16711622 TI - The seven basic virtues in medicine II. Justice. PMID- 16711623 TI - Intensive care ventilators. AB - This study presents ECRI's evaluation findings for four newly tested intensive care ventilators, as well as updated ratings for six previously evaluated models. Intensive care ventilators provide temporary support for critically ill patients who cannot breathe on their own or who require assistance to maintain adequate gas exchange in the lungs. Most of today's models offer a variety of capabilities to help clinicians tailor treatments to each patient, to alert users to potentially dangerous conditions, and to protect the patient when problems arise. We found that all the evaluated models can satisfactorily ventilate patients. However, not all the models would be an appropriate choice for the typical intensive care unit (ICU). Some of the units we tested lack capabilities that we desire for intensive care applications, and some include noteworthy deficiencies, particularly related to their alarm systems. In fact, we rate one unit Unacceptable because its alarm limits default to unsafe values. Of the remaining units, we rate five models Preferred because they offer the functionality to meet the needs of a broad range of ICU patients. Two additional units offer more limited capabilities, but they perform well and are inexpensive; these models, which we rate Acceptable, may be the best choice for some facilities. The final two units are Not Recommended for most new purchases because they lack features that aid in patient-ventilator synchrony; such features can improve patient comfort and thus are highly desirable. ECRI's recommended purchasing strategy, which is detailed in the Conclusions section, will help healthcare facilities select from among the evaluated models. PMID- 16711624 TI - Trailing zeros on bedside monitor drug dose calculators contribute to dosing errors. PMID- 16711625 TI - Longitudinal study of non-cavitated carious lesion progression in the primary dentition. AB - OBJECTIVES: This study reports changes in non-cavitated tooth surface diagnoses after a 4-year period. METHODS: Dental examinations were conducted for Iowa Fluoride Study cohort children who had non-cavitated lesions in the primary dentition and were also examined an average of 4 years later in the mixed dentition. Comparison of fluoride exposures, socioeconomic factors, and beverage consumption patterns were made between children who had lesions progress and those who did not. RESULTS: Of 129 non-cavitated pit and fissure lesions in the first exams, 40 (31%) progressed to either frank decay or filled status, while among 132 noncavitated smooth surface lesions, 7 (5%) were filled and none had frank decay in the second exam. No fluoride, socioeconomic status or beverage variables were significantly associated with lesion progression. CONCLUSIONS: Non cavitated smooth surface lesions rarely progressed in this age group, but nearly one-third of pit and fissure lesions progressed. PMID- 16711626 TI - Dental caries experience and factors among preschoolers in southeastern Mexico: a brief communication. AB - OBJECTIVE: To examine the association between dental caries prevalence and selected variables in preschool children. METHODS: A cross-sectional study was carried out with 1,303 preschoolers (ages 3-6 years old), and the mothers completed questionnaires. The children were examined by one of three standardized dental examiners. Logistic regression was performed to identify associations between dental caries and other factors. RESULTS: Mean dmft was 1.54+2.47, with 44.1% of children having dmft>0. Caries prevalence was associated with older children (OR=1.39); medium (OR=1.66) and low (OR=2.41) socioeconomic levels; mediocre (OR=1.71) and inadequate (OR=2.25) hygiene; negative attitude toward oral health (OR=1.51); and the presence of enamel defects (OR=1.74). CONCLUSION: Both overall caries prevalence and dmft index were relatively low. The results of this study substantiate previous reports in the international literature for clinical, behavior, socio-demographic, and socio-economic variables that contribute to dental caries in Mexican children. PMID- 16711628 TI - Determining dental utilization rates for children: an analysis of data from the Iowa Medicaid and SCHIP programs. AB - OBJECTIVE: Accessing dental care is a significant problem for children in Medicaid and SCHIP. Evaluating the extent of the access problem is affected by the differential dental utilization rates as calculated by unique approaches used by national organizations. The problem is related to the intermittent enrollment in Medicaid and SCHIP for many children during the year. The objective of this study is to evaluate the effect of four different approaches for calculating dental utilization rates for children in Medicaid and SCHIP programs. METHODS: All Iowa Medicaid and S-SCHIP dental claims and enrollment files for CY 2001 were used to characterize the populations, calculate dental utilization rates and evaluate the types of services received by dental utilizers. Dental utilization rates were calculated four ways using the following different rate denominators: 1) any child enrolled during the year (Center for Medicare and Medicaid Services method), 2) children enrolled for 11-12 months (National Committee for Quality Assurance method), 3) a full-time equivalents (FTE) method, 4) only newly enrolled children. RESULTS: The methodology employed greatly affected the dental utilization rates. Rates varied from 18% for newly enrolled children in Medicaid to 58% for S-SCHIP-enrolled children using the FTE method. Methods that included children who were more likely to be in for more months during the year, such as the NCQA approach, produced the highest rates. CONCLUSIONS: The method used to determine the dental utilization rates for Medicaid and SCHIP enrollees should be clearly stated when these rates are being presented. This will allow the reader to be able to make a careful and appropriate interpretation of the results. PMID- 16711627 TI - Use of the fluorosis risk index in a cohort study: the Iowa fluoride study. AB - OBJECTIVES: Several studies have used the Fluorosis Risk Index (FRI) to assess risk factors for dental fluorosis. This study reports fluorosis prevalence estimates in a birth cohort using the FRI in different ways. METHODS: Subjects (n=443) were participants in the Iowa Fluoride Study, a birth cohort study of fluoride exposures and intake, fluorosis, and caries. Early-erupting permanent teeth were assessed for fluorosis using the FRI at approximately age 9. Fluorosis prevalence rates were calculated separately for specific teeth and different combinations of teeth, with varied FRI zones included and using different thresholds. Mean numbers of teeth with fluorosis were calculated. RESULTS: Tooth specific fluorosis prevalence varied, with maxillary central incisor fluorosis most prevalent and mandibular incisors least affected. Considering three zones of each tooth (cervical zones excluded), 40.6% overall had at least one tooth with mild or more involved fluorosis, 30.2% were questionable fluorosis, and 29.1% had no fluorosis. When only FRI zone I areas were considered, the corresponding percentages were 33.2%, 29.3%, and 37.5%, respectively. When different combinations of teeth were used to define fluorosis cases, the prevalences estimated using three zones were generally 1 to 9 percentage points higher than those estimated from FRI zone 1 only. Most fluorosis was mild, with only 7 individuals (1.6%) having FRI severe fluorosis. CONCLUSIONS: The FRI has advantages for use in analytical epidemiologic studies of dental fluorosis. However, the population prevalence estimates vary, depending on the index and case definition used. Therefore, it is recommended that consideration be given to concurrent use of another index (i.e., TSIF, TF, Dean's) if prevalence estimates are an important study outcome. PMID- 16711629 TI - Adult oral health inequalities described using area-based and household-based socioeconomic status measures. AB - OBJECTIVES: To describe adult oral health inequalities using an area-based and household-based measure of socioeconomic status (SES). METHODS: Self-report questionnaires (seeking information on sociodemographic, oral health and oral self-care) were sent to a random sample of adults from the Dunedin South Electorate, New Zealand. Household- and area-based SES measures were collected. The main outcome measures were edentulism prevalence, average-poor self-rated oral health and not having visited a dentist for 2+ years. Data were weighted to produce population-based estimates. RESULTS: The response rate was 78.2%; the sample mean age was 47 years (sd, 17; range 18-92 years) and females comprised 54.0%. Edentulism was most prevalent among those from low-SES households who were resident in high-deprivation areas (P<0.0001). Poor self-rated oral health (P<0.0001) and 2+ years since the last dental visit (P<0.0001) were also most prevalent among these same individuals. In contrast, respondents from high-SES households located in the least deprived areas had the lowest prevalence of edentulism, poor self-reported oral health or 2+ years since their last dental visit. Those from the other household/area SES combinations occupied intermediate positions. CONCLUSIONS: There may be added value to dental public health in using a dual socio-economic measurement approach to population research, with greater oral health gains perhaps being possible by concentrating resources and clinical effort on people living in low-SES households in highly-deprived areas, rather than those living in low-SES households in areas that are not deprived. PMID- 16711630 TI - Tooth loss in a young population from south Brazil. AB - OBJECTIVES: To assess the prevalence, extent, and risk indicators of tooth loss in a representative young urban population from south Brazil. METHODS: A representative sample was drawn using a multi-stage probability cluster sampling strategy, and consisted of 612 subjects 14-29 years of age in the metropolitan area of Porto Alegre, Brazil. A clinical examination was carried out by 4 calibrated examiners in a mobile examination center. RESULTS: The prevalence of tooth loss was 44.8%, 26%, and 60%, and the mean tooth loss was 1.4, 0.6, and 2.4 teeth in the age groups 14-29, 14-19 and 25-29 years, respectively. First molars were the most frequently missing teeth, and the mandibular incisors and canines were the least missing teeth. Tooth loss increased sharply with age, and was similar in males and females. Having > or =4 missing teeth was significantly associated with low socioeconomic status and heavy smoking, and was significantly more likely in persons who had > or =2 teeth with caries/fillings and/or > or =5 mm attachment loss. CONCLUSION: Tooth loss is a dental health concern in this young Brazilian population. Community-based oral diseases prevention programs targeting groups having these risk factors should be implemented to reduce tooth loss. PMID- 16711632 TI - The role of location in indigenous and non-indigenous child oral health. AB - OBJECTIVE: To examine the role of location in Indigenous and non-indigenous child oral health in three Australian states and territories. The association of Indigenous status and residential location with caries prevalence, severity and unmet treatment need was examined. METHODS: Data were collected as part of a national monitoring survey of 4-14-year-old children enrolled in school dental services in New South Wales, South Australia and the Northern Territory, Australia. RESULTS: Of the 326,099 children examined, 10,473 (3.2%) were Indigenous. Fewer 4-10-year-old rural Indigenous children were caries-free in the deciduous dentition than their non-indigenous counterparts and rural Indigenous children had almost twice the mean number of decayed, missing and filled teeth (dmft) of rural non-indigenous children. The % d/dmft was higher among rural Indigenous children than rural nonIndigenous children. Fewer 6-14-year-old rural Indigenous children were caries-free in the permanent dentition than their non indigenous counterparts and rural Indigenous children had almost twice the mean DMFT of rural non-Indigenous children. The % D/DMFT was higher in rural Indigenous than rural non-indigenous children. Living in a rural location was the strongest indicator of caries prevalence, severity and unmet treatment need in the deciduous dentition of Indigenous 4-10-year-olds while being socially disadvantaged was the strongest indicator of poor oral health outcomes among older Indigenous and all non-Indigenous children. CONCLUSIONS: Living in a rural location exhibited the strongest association with poor oral health outcomes for young Indigenous children but was also associated with poorer oral health among older Indigenous and non-Indigenous children. PMID- 16711631 TI - Is regular visiting associated with lower costs? Analyzing service utilization patterns in the first nations population in Canada. AB - OBJECTIVES: Using an administrative database of dental service records from the Non-Insured Health Benefits (NIHB) program of Health Canada for 1994-2001, the authors set out to test whether regular visitors had lower program expenditures. METHODS: The age-specific mean expenditures per client were compared among those with regular examinations in 8, 7 and fewer years. The study further examined the effect of regular visiting over the first 6 years on expenditures in the last 2 years. "Continuity of care" was measured by the numbers of consecutive years prior to 2000 in which clients had a regular examination. In a "gap analysis" individuals were classified according to the number of years prior to 2000 since they last had an initial or recall examination. Mean expenditures per client were analyzed by age group and type of service. FINDINGS: Over the 8-year period, clients with regular visits had the highest expenditures. In both the continuity of care and gap analyses, the findings were generally consistent; the more that clients visited over the first 6 years, the higher the expenditures in the final 2 years. Clients with more "regular" (initial and recall) examinations received a relatively standard, age-specific, pattern of service but incurred greater expenditures compared to clients with fewer regular, or longer gaps in, examinations. CONCLUSION: The observations of the authors in this client group do not support the thesis that regular visiting is associated with lower expenditures on dental care. PMID- 16711633 TI - Oral health status of mississippi delta 3- to 5-year-olds in child care: an exploratory study of dental health status and risk factors for dental disease and treatment needs. AB - OBJECTIVES: For poor and minority young children, disparities exist in dental health and treatment. In rural impoverished areas, institutions that reach young children and potentially offer access to care are limited. In the current Mississippi Delta study, child care centers were examined as potential venues for oral health intervention and research, and potential risk factors for dental caries and treatment urgency in high-risk preschool children were explored. METHODS: Child care centers were selected and attending children recruited. Data on oral health practices were collected from surveys of center directors and parents/caregivers. Children were examined for caries and treatment urgency at centers by dentists. Bivariate and multivariate analyses with a 0. 05 alpha were used to examine data. RESULTS: A total of 346 preschool children at 15 participating centers were examined: 46% were female, 68% minority. Minority children and those with public insurance were more than twice as likely to have caries and urgent treatment needs as non-minorities or those with private insurance. The odds of children having caries were half as great if parents reported using floss and nearly twice as great if the parent had experienced a dental abscess. For every soft drink the parent consumed daily, the odds of dental caries for children increased by 44%. CONCLUSIONS: Conducting oral health exams and research in child care venues was possible, yet presented challenges. The combined use of two parental variables, reported soft drink consumption and abscess history, appears promising for caries prediction. Implementation of oral health programs and research in child care venues merits further exploration. PMID- 16711634 TI - Role of african-american fathers in child-rearing and oral health practices in an inner city environment--a brief communication. AB - OBJECTIVE: This study assesses parenting behaviors, health behaviors and attitudes in a sample of disadvantaged African-American fathers and evaluates the potential of fathers as recipients of oral health promotion interventions. METHODS: Participants were 60 African-American fathers whose children were enrolled in a childhood caries prevention study at Women and Infant Child Program (WIC) (n=25) or who were members of a Young Fathers' Program (n=35) at an urban health center. Demographic factors, oral health knowledge, dental self-efficacy, parenting behaviors, readiness to change, and health promotion behaviors were assessed. RESULTS: Participants averaged 30.8 years of age (sd=8.6); children averaged 2.1 years (sd=0. 9). Half of the fathers always have lived with their own children and 92% had regular physical contact with their children. Almost half of the sample reported 'fair/poor' oral health status, while less than 25% reported 'fair/poor' general health. Fathers had high oral health knowledge scores (x=7.7 correct of 9 items), and high readiness for change scores at the action stage (X=12.2 of 15; sd=1.9). dental self-efficacy scores were moderate (X=15.7 of 24; sd=5.5). Half of the fathers shared or had sole responsibility for brushing their children's teeth; 90% report children's teeth were brushed daily. CONCLUSIONS: African- American fathers participated in childrearing and oral health promotion behaviors. PMID- 16711635 TI - Prevalence and causes of oral injuries in a population of Canadian adults aged 18 to 50 years--a brief communication. AB - OBJECTIVE: To determine the prevalence and causes of injuries to the mouth and teeth among adults aged 18 to 50 years in Ontario, Canada. METHODS: 2001 individuals in the target age range participated in a telephone interview survey based on random digit dialling. RESULTS: Overall, 15.5% of subjects reported a history of injury with 5.4% having multiple episodes of injury. One third of the injuries occurred in adulthood. The majority (85.0%) with a history of trauma reported damage to the teeth and of these, 26.0% broke one or more teeth, 25.4% reported one or more teeth being knocked out and 6.5% reported displacement of teeth. The most common cause of injury was playing sports, followed by collisions and falls. Violence accounted for 10.0% of the injuries reported. The causes of injury varied according to gender, education and the age at which the injury occurred. CONCLUSION: Oral and dental injuries occur throughout the lifespan, although the causes appear to vary according to the age at which they happen. PMID- 16711636 TI - Dental anxiety among young Israeli male adults as related to treatment received during childhood. AB - OBJECTIVES: To evaluate dental fear and anxiety among young male adults treated during childhood either by a certified pediatric dentist or a general dental practitioner. METHODS: A structured questionnaire that included information regarding issues related to dental fear and anxiety (Intake Interview Questionnaire and the Dental Anxiety Scale) was completed by 497 young male army recruits (18 years old). RESULTS: During childhood, 59 (11.9%) of the participants were treated on a regular basis by a certified pediatric dentist and 344 (69.2%) by a general dental practitioner. Sporadic care (not regular care) by general dental practitioners was given to 94 participants (18.9%). Average level of dental anxiety among participants treated by certified pediatric dentists was 7.9 compared to 8.2 (general dentists) and 8.7 (sporadic care by general dentists). There were no significant differences between the groups. However, significant differences were found in the prevalence of high dental anxiety between participants treated by a regular dentist in childhood and those treated sporadically (15.4% vs. 23.4%, respectively). Negative emotions were common. Environmental etiologic factors were infrequently reported. CONCLUSIONS: The general level of dental anxiety among young Israeli males was normal. No relation was found between level of dental anxiety and the professional education of the treating dentist in childhood. Regular dental care in childhood has more effect on preventing the development of high dental anxiety than the professional education of the early caregiver. PMID- 16711637 TI - Prevalence of xerostomia in population-based samples: a systematic review. AB - The consequences of xerostomia on oral health have been studied for decades; however, the actual prevalence of this disorder on the general population remains controversial. The purpose of this systematic review was to determine the prevalence of xerostomia in population-based samples. Electronic databases were screened for relevant articles and reference lists of pertinent articles were also hand-searched. Thirteen articles meeting the final inclusion criteria were identified. Based on the definition of xerostomia used in this review, only publications including a subjective diagnosis of the disorder were included. All of the self-reported diagnoses were achieved through a questionnaire either by mail, telephone, interview or self-administered by the patient. There was a variation across papers regarding number and content of the questions as well as guidelines for the diagnosis of xerostomia (i.e. answer yes to one or more than one question). The prevalence of xerostomia in the selected articles ranged from 0.9% to 64.8%. The majority of these studies were performed in Scandinavia. In 9 of the articles, the studied samples were 50 years and older. None of the studies evaluated the prevalence xerostomia among individuals younger than 18 years. Based on these observations, it can be concluded that there is a need for population-based studies on prevalence of xerostomia in regions other than Scandinavia. A standardized protocol to diagnose xerostomia needs to be developed. Patient-perceived treatment needs and impact on quality of life should be included to have a complete picture of public health implications of the disease. PMID- 16711638 TI - [Ethical analysis of the decision-making process in occupational health practice]. AB - BACKGROUND: Changes in workplaces and work organizations represent a challenge for governments, social partners and occupational health professionals whose aim is to appropriately satisfy emerging requirements and needs. An increasing number of occupational health problems requires a high-quality standard practice supported by ethically consistent decisions. The ethical aspect of the practice is strictly linked to that of appropriateness, involving requirements of effectiveness, efficiency and respect of ethical principles of the individual, community and society. OBJECTIVES AND METHODS: The paper aims at focusing the ethical components of the quality of an occupational health program by taking a systematic approach to the ethical problems. The approach consists of a 2-step process. The first step consists of appraising the basic ethical principles of the dilemma ("to do good": to prevent or to remove evil and to do or to promote good; "not to do evil", implying not to inflict evil; to do no harm; autonomy implying respect of other's freedom and self-determination; justice implying equity, solidarity and non discrimination). The second step consists of detecting the stakeholders involved or interested in the decision. The alternatives are discussed according to the assessment of ethical costs (violating the consistentprinciple) and ethical benefits (fulfilment of the consistent principle) for the stakeholders. RESULTS: Systematic analysis of the ethical components of the dilemma according to ethical principles and their discussion within a framework involving different stakeholders makes it possible to recognise ethical costs and ethical benefits of the alternative decisions. The decisions may have different costs and benefits, which should be considered and weighed to take a proper decision. Although there is no certainty about the suitability of the decision, the assessment of the ethical components may be a valuable tool in decision-making based on the awareness that any ethical aspect has been considered. CONCLUSION: Occupational health professionals are requested to act with respect for general ethical principles and preferences of the individual, groups and setting. A high-quality practice will take into account the ethical content and the ethical conduct and any intervention will include analysis of ethical principles to compare alternative decisions and their consequences for the different stakeholders. This approach guarantees a practice based on, in addition to scientific evidence, the assessment of ethical costs and ethical benefits to favour decisions preventing conflicts. PMID- 16711639 TI - [Fifty years of impact factor: pros and cons]. AB - BACKGROUND: The Impact Factor (IF) was thought up 50 years ago, and it is currently the best available bibliometric index. Despite its shortcomings, the IF is widely used and offers, at present, the best simple tool to help libraries decide which journals to purchase. DISCUSSION: The use of the term "impact factor" has gradually evolved, especially in Europe, to include both journal and author impact. With this meaning, it has become the most used tool in evaluating scientific quality in decisions regarding funding and academic career. This ambiguity and use of IF to compare authors may be misleading. There is poor correlation between the citation frequency of a certain article and the IF of the journal in which it is published. There is a marked bias towards the English language journals compared with those in other languages. Even within the field of Occupational Medicine, the distribution of IF exhibits a wide spread between specialty subsets. CONCLUSION: The IF is a flawed measure of quality of academic staff performance. Searching for the highest IF is not equal to searching for the truth. There is still much work to be done to develop truly objective measures of scientific quality. Like all measures, the use of IF has to be tempered with knowledge of its limitations and common sense. PMID- 16711640 TI - [Environmental and biological monitoring of exposure to polycyclic aromatic hydrocarbons in workers of an electric steel foundry]. AB - AIMS: To assess and classify exposure to Polycyclic Aromatic Hydrocarbons (PAHs) in some specific working areas of a steel foundry operating with a continuous casting process and evaluate biomonitoring data in different job tasks. METHODS: Exposure to dusts and six PAHs classified as carcinogenic by EU directives was studied in a cohort of 35 male foundry workers (aged 41.1 +/- 6.9 years), who were examined both prior to and at the end of the work-shift (06:00 a.m.-02:00 p.m.) in two different periods. The urinary excretion of 1-hydroxypyrene (1-OH-P) was measured as a biomarker of exposure to pyrene. RESULTS: PAHs concentrations ranged from 461.8 to 935.6 ng/m3 near the continuous casting area, whereas lower values were measured near the ladle furnace. End of shift 1-OH-P values were higher in 11 non-smoking workers involved in continuous casting process as compared to those employed in mantenance and furnace areas (median of the second determination: 5.70 microg/g creatinine--range: 1.24-21.24 vs 1.17 microg/g creatinine--range: 0.23-4.49; p< 0.001). 1-OH-P excretion was significantly correlated with both the sum of six carcinogenic PAHs and pyrene airborne concentrations. In two biomonitoring sessions, 9.1% and 34.3% of the workers respectively showed end-of-shift 1-OH-P values exceeding the occupational exposure limit (OEL) (4.4 microg/g creatinine or 2.3 micromol/mol(-1) creatinine) recommended for coke-oven workers. CONCLUSIONS: 1-OH-P is a useful biomarker in assessing PAH exposure and is associated with job category at a Steelplant. Due to exposure variability, to assess risk associated with PAHs exposure, biological monitoring should be carried out periodically. PMID- 16711641 TI - [Educational and training issues in occupational first aid: the requisites for the teachers of the workers responsible for first aid]. AB - BACKGROUND: The inter-ministerial decree (D.I.) 388/03 (the Italian law on first aid in the workplace) attaches particular importance to the organization of first aid and to the content of the teaching and training of designated workers, specifying teaching objectives, programme and duration of courses. However it provides less details on the requirements and qualifications of the teachers and trainers. OBJECTIVES: To discuss this topic in the light of the guidelines contained in the literature and the existing legislation. METHODS: We used the specific references of literature and legislation concerning first aid training of designated workers. RESULTS: The objectives and the content of the training courses call for a high level of professional competence and approach. Therefore, training of designated workers should be planned and carried out in accordance with the training standards used for the teaching and training of the so-called "lay rescuers". CONCLUSIONS: The need to achieve high training standards for workers responsible for first aid at the workplace is to a large extent justified by the difficulty of the role of employees. This need follows the current tendency in regulations to guarantee adequate and up-dated training also for other agents operating within the system of workplace prevention. In this context the occupational physician plays a significant role in the training programmes for designated workers. PMID- 16711642 TI - [Occupational exposure to nickel in Tuscany: analysis of a biological monitoring register for the period 1991-1998]. AB - OBJECTIVE: The study assesses the time trend in exposure to nickel among factory workers in Florence, via data on biological monitoring. A data-base of nickel in urinary samples (Ni-U mg/l) was created for the period 1991 to 1998. METHODS: The data-base contained 2.138 samples, measured by atomic absorption (GF-AAS),from 893 workers. Subjects came from 157 factories in various manufacturing sectors, especially electroplating, mechanical workshops, jewellery. RESULTS: Ni-U levels were correlated with manufacturing sector. The highest levels were found among workers from electroplating industries, where exposure was mainly due to water soluble nickel compounds. The eight-year time trend showed a statistically significant decrease in Ni-U values, with a sharper drop during the last two-year period. Age, sex and number of samples per subject were not statistically related to this trend. CONCLUSION: The observed Ni-U decrease could be related to the efficacy of new legislation introduced in Italy during the study period (Law 626/94 and subsequent laws), but also to the intense labour inspection activities that officials of National Health Service performed, which were rightly focused on nickel exposure in different manufacturing sectors. This study confirms the usefulness for occupational risk evaluation of a biological monitoring data-base of routinely collected data. PMID- 16711643 TI - [Photoaging of the skin and occupation: correlation between clinical ultrasound and histological findings]. AB - OBJECTIVES: We described the ultrasound (US) signs that can define occupational photoaging of the skin. We also studied the clinical importance of these signs, so as to aid the specialist in the diagnosis and evaluation of "anti-aging" treatments. PATIENTS AND METHODS: The dermis and hypodermis of 180 subjects (6 groups of 15 men and 15 women each), were studied with US test (echograph Sonora 400 MD). We used a 10 MHz probe and recorded the thickness and echogenicity of the dermis and hypodermis. US images were correlated with the histological findings of 60 patients. RESULTS: In skin not exposed to sunlight aging produces an increase in thickness and a reduction in echogenicity, whereas in photoaging there is a reduction in thickness and an increase in echogenicity of the skin. CONCLUSION: Photoaging produces an increase in elastic fibres and consequently hyperechogenicity of the dermis, while, in the advanced phase, the loss of collagen and elastic fibres justify the reduction in thickness and echogenicity of the dermis. These observations allow as to classify the US signs of the photoaging of the skin into four clinical/ultrasound stages. PMID- 16711644 TI - [Two cases of pleural mesothelioma following unusual and unrecognized exposure to asbestos. The role of Occupational Health and Safety Service in identifying past occupational exposure]. AB - BACKGROUND: In Italy there was a wide use of asbestos in various manufacturing sectors and for many different uses, some of which are still partly or completely unknown. A detailed reconstruction of the work histories of mesothelioma patients made it possible, in some cases, to identify ignored circumstances of asbestos exposure. Moreover, the identification of cluster of cases takes on special significance in suggesting a possible previous asbestos exposure, where the information collected on single cases do not imply as much. OBJECTIVES: This report concerns two cases of malignant mesothelioma that occurred in two workers employed in the same processes in a small factory that manufactured and repaired electric motors for hand tools. METHODS AND RESULTS: In the Province of Brescia (one million inhabitants) a Mesothelioma Register is in operation. The first case was classified, according to Re.Na.M.1996 criteria (National Mesothelioma Register) as "unknown" occupational exposure. The identification of a second case, that was discovered thanks to the surveillance system of the Mesothelioma Register, encouraged the local Occupational Health and Safety Service to perform a more detailed investigation that revealed, for both subjects, previously unknown occupational exposure. This consisted of grinding, in a damp setting, electric motor parts bushed with phenolic thermosetting resins reinforced with chrysotile asbestos. Moreover, weekly cleaning of the plants could have been an occasion for dust dispersion. It is likely that this exposure did not last long and was limited in extent. Other similar reports of such circumstances of occupational exposure were not available in the literature. CONCLUSIONS: The results confirm the high information value of systematic collection of incidental cases in the population, which is feasible thanks to the disease register, and the significant role of the local Occupational Health Services in demonstrating past asbestos exposure. PMID- 16711645 TI - [Psychiatric aspects in workers subject to mobbing]. AB - BACKGROUND: Mobbing in the language of ethologists means a particular behaviour of a group of animals consisting of several members forming an alliance against a single animal, in order to eliminate this animal away from the group. This term has been adopted in current usage to mean a kind of relationship at the workplace that, by aggressive and underhand means, aims at influencing or expelling the person. The term "bossing" is used to indicate a negative action, under the meaning described above, made by one or more senior persons, or possibly by the employer. In the Italian Parliament, under the current legislature, several bills are pending regarding this problem. Besides the aspects of topical interest in the problem, there are first and foremost the different kinds of reaction adaptation to mobbing, according to the type of individual internal working models in action, that can be anxious/avoiding, anxious/resistant or disorganised. OBJECTIVES: The purpose of our study was a clinical evaluation of the different ways of adaptation-reaction to a mobbing-type relationship at the workplace, and of the type of internal working models found among the examined patients. METHODS: A clinical interview was carried out with 40 patients who, over a period of 12 months of observation, came to request a clinical assessment, for medical-legal reasons, of their psycho-pathological state and ways to react to adverse situations at the workplace, such as a mobbing or bossing. RESULTS: Besides the psychological and relational effects of mobbing, the reactions of the examined subjects to mobbing are also described, considering also the different cognitive self-organization possibilities, according to the individual internal working model, regarding patterns of attachment in adult age. The most frequent internal working model seems to be the one related to resistant attachment, followed by the avoidance and lastly--rarely--the disorganized model. Furthermore we observed among these subjects a certain deficit in meta-cognitive capacity of self reflection and decentralization. CONCLUSIONS: Cognitive therapy is proposed as the most effective of therapeutic approaches, associated with symptomatic psycho-pharmacological therapy as needed. Initial clinical results on cognitive therapy of these patients are reported. The individual type of internal working model seems to be the critical element in mobbing reaction and, besides specific intervention plans regarding relationship in the workplace, individual psychotherapy appears to be the best instrument of treatment and prevention of the psychopathologic effects caused by mobbing. PMID- 16711646 TI - [Cluster cases of malignant pleural mesothelioma in an oil factory]. AB - BACKGROUND: The traditional occupational hazards of the productive cycle of oils are attributable to chemicals (use of solvents, pesticides and other agents), dusts, labour accidents (trauma, ignition, explosion), noise, manual lifting, work organization and hot-wet microclimate. The latest risk is due to the use of high temperatures (from 50 up to 250 degrees C) during the processes of extraction with solvent and refining. No cases are reported in literature of asbestos related disease in subjects who worked in oil factories. Nevertheless the structure and organization of the workplace, which is similar to that of sugar refineries, where cases of malignant mesothelioma have been described (moreover in workers employed in running and maintenance of the plants), led to the assumption that even in oil factories asbestos for the insulation of pipes and boilers could be present. OBJECTIVES: To describe 3 cases of Malignant Mesothelioma that occurred in workers of the same oil factory. METHODS: Since this occupational sector is not conventionally known for asbestos exposure the Local Health Unit and the Lombardy Mesothelioma Registry decided to investigate this industrial plant. RESULTS: Following examination of the archives of the Local Health Unit and inspection of the plant, an environmental asbestos contamination (pipes and boilers) was found. The 3 cases were defined as occupational disease and the required legal procedures were initiated. This underlines the importance of close cooperation with Local Health Units of occupational medicine and the Regional Mesothelioma Registry in the study and acknowledgment of cases which would otherwise not have been recognized, with consequent loss of precious information. PMID- 16711647 TI - [A methodological question concerning the evaluation of hearing loss due to occupational noise]. PMID- 16711648 TI - Commentary to the paper by P. Bernardini et al "Malattie mieloproliferative da uso di benzina come solvente: descrizione di tre casi" Med Lav 2005; 96: 119-125. PMID- 16711649 TI - [Profiles of university occupational medicine teachers. Document of the Italian Society of Occupational Medicine and Industrial Hygiene, the B. Ramazzini National University Section]. PMID- 16711650 TI - Vitamins A and E status in an urban Lebanese population: a case study at Dar Al Fatwa area, Beirut. AB - Vitamin A and E status are widely studied in various populations because of their association with several diseases. Fasting plasma vitamin A and E status of 857 Lebanese adults residing in Dar Al-Fatwa, Beirut were assessed using high performance liquid chromatography (HPLC). Mean retinol and alpha-tocopherol concentrations were 59.8 +/- 29 microg/dL and 1.0 +/- 0.5 mg/dL, respectively in which only 0.2% were retinol-deficient, while 0.7% were alpha-tocopherol deficient. Vitamin A and E correlated positively with plasma concentrations of total cholesterol, low-density lipoprotein (LDL)-cholesterol, and triglycerides. Vitamin E, but not vitamin A, correlated positively with blood pressure and glucose. A good status of vitamins A and E was found among the studied Lebanese sample and the elevation in vitamin A and E levels was associated with unfavorable lipid profile. PMID- 16711651 TI - Cloning of the bovine beta-carotene-15,15'-oxygenase and expression in gonadal tissues. AB - Beta-carotene-15,15'-oxygenase (betaCO), found mainly in intestinal mucosa and liver, is the enzyme responsible for cleaving beta-carotene into retinal, which can be used or stored at these sites or carried by the bloodstream to different target cells within the body. We isolated the cDNA for bovine betaCO and demonstrated its expression in gonadal tissues. A cDNA of 2130 base pairs (bp) was obtained by reverse transcriptase-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE), using degenerate oligonucleotides; the deduced protein shared an identity of 75% with its homologues from other mammalian species. In order to evaluate the expression of this enzyme, we performed RT-PCR and in situ hybridizations in the ovary and testis of bovines. RT-PCR showed the expression of betaCO in testis, ovary, and cultured granulosa cells. In situ hybridization of complete ovary and testis revealed expression in granulosa cells and the corpus luteum in the ovary and in germinal and interstitial cells in the testis. These results suggest that beta-carotene could act as a local source of retinoids, which have been shown to be important during proliferation, differentiation, and maturation of both female and male germinal cells. PMID- 16711652 TI - Alpha-linolenic acid content of commonly available nuts in Hangzhou. AB - The total lipid content of eight species of nuts available in Hangzhou ranged from 49.5 g/100 g weight in Cannabis sativa to 75.4 g/100 g in walnut. The predominant content of lipid is triacylglycerol, ranging from 91.1% in Cannabis sativa to 98.4% in macadamia. There were two polyunsaturated fatty acids (PUFA) in all nuts analyzed; 18:2n-6 and 18:3n-3. The content of 18:3n-3 ranging from 0.2% in almond to 15.2% in Cannabis sativa, 18:2n-6 ranged from 2.5% in macadamia to 61.6% in pine nut. The proportion of total PUFA in analyzed eight nut species ranging from 2.8% in macadamia to 71.7% in walnut (p < 0.001). Monounsaturated fatty acid composition ranged from 18.0% in Cannabis sativa to 82.6% in macadamia (p < 0.001). The proportion of saturated fatty acid ranged from 7.4% in filbert to 14.7% of total fatty acids in macadamia (p < 0.001). No C20 fatty acids were detected in any of the samples in the present study. The lipids content and fatty acid compositions in analyzed samples were varied between nut species. Cannabis sativa and walnut contained relatively high 18:3n-3, consumption of several these nuts each day can contribute to n-3 PUFA intake, especially for the vegetarian population. PMID- 16711653 TI - In vivo antioxidant activity of procyanidin-rich extracts from grape seed and pine (Pinus maritima) bark in rats. AB - BACKGROUND: In vitro evidence exists for the potential antioxidant benefits of procyanidin-rich extracts, but in vivo studies are scarce. We have evaluated the effects of selected procyanidin-rich extracts on oxidative stress in rats in condition of prolonged consumption of these compounds and also after single administration i.e. in postprandial conditions. METHODS: Rats were fed for 8 weeks with diets supplemented with either a grape seed extract (GE), a pine bark extract (PE), or a high-degree polymerized pine bark extract (HPE). An additional study was performed in order to assess the postprandial effect of these extracts on plasma antioxidant capacity. The ferric-reducing antioxidant power (FRAP) and thiobarbituric acid-reactive substances (TBARS) were determined in plasma. For lipid peroxidation study of heart tissue, homogenates were prepared and TBARS were measured after lipid peroxidation induced by FeSO4-ascorbate. RESULTS: After 8 weeks of dietary treatment, total antioxidant capacity in plasma was significantly higher in the GE and PE groups as compared with the other two groups. Plasma TBARS concentrations and heart susceptibility to peroxidation were not significantly different between the groups. In the postprandial state, by comparing plasma antioxidant capacity 2 hours after ingestion of the different procyanidin-rich extracts (500 mg/kg body weight), we observed that FRAP values were higher in the procyanidin-rich extracts groups as compared with the control group. Moreover, plasma FRAP concentration was significantly higher in the GE group as compared with the other groups. CONCLUSION: The results of the present experiment constitute positive evidence for an in vivo antioxidant effect at the plasma level of procyanidin-containing plant extracts. PMID- 16711654 TI - Tryptophan-niacin metabolism in rat with puromycin aminonucleoside-induced nephrosis. AB - We investigated the change of tryptophan-niacin metabolism in rats with puromycin aminonucleoside PAN-induced nephrosis, the mechanisms responsible for their change of urinary excretion of nicotinamide and its metabolites, and the role of the kidney in tryptophan-niacin conversion. PAN-treated rats were intraperitoneally injected once with a 1.0% (w/v) solution of PAN at a dose of 100 mg/kg body weight. The collection of 24-hour urine was conducted 8 days after PAN injection. Daily urinary excretion of nicotinamide and its metabolites, liver and blood NAD, and key enzyme activities of tryptophan-niacin metabolism were determined. In PAN-treated rats, the sum of urinary excretion of nicotinamide and its metabolites was significantly lower compared with controls. The kidney alpha amino-beta-carboxymuconate-epsilon-semialdehyde decarboxylase (ACMSD) activity in the PAN-treated group was significantly decreased by 50%, compared with the control group. Although kidney ACMSD activity was reduced, the conversion of tryptophan to niacin tended to be lower in the PAN-treated rats. A decrease in urinary excretion of niacin and the conversion of tryptophan to niacin in nephrotic rats may contribute to a low level of blood tryptophan. The role of kidney ACMSD activity may be minimal concerning tryptophan-niacin conversion under this experimental condition. PMID- 16711655 TI - Effects of short-term supplementation with ascorbate, folate, and vitamins B6 and B12 on inflammatory factors and estrogen levels in obese postmenopausal women. AB - Little is known about the effects of commonly used vitamins on serum inflammatory markers and the hormonal balance in obese postmenopausal women. We studied the effects of an 8-week open-label supplementation with vitamins C (500 mg), B6 (25 mg), B12 (1 mg), and folate (5 mg) on C-reactive protein, interleukin-6, and estradiol levels in 20 obese (body mass index > or = 30) postmenopausal women. Outcomes were assessed in a blinded fashion. Folate and vitamin B12 levels rose significantly, suggesting that the supplement was well absorbed and that participants adhered to the protocol. Weight, blood pressure, and serum lipids remained stable. C-reactive protein, interleukin-6, and leptin levels remained unchanged. Estradiol levels rose from a median of 22.0 pg/mL (IQR = 15.9-25.8) at baseline to a median of 27.8 pg/mL (IQR = 23.1-33.9) at follow-up (p = 0.003). Increments in serum estradiol caused by vitamin supplementation in postmenopausal women have not been previously described and probably merit further investigation. PMID- 16711656 TI - Influence of training status on glycemic index. AB - The glycemic index (GI) represents the relative postprandial blood glucose response to the ingestion of a food containing carbohydrate. Although regular physical exercise may influence glucose metabolism, it is not yet known if chronically performed exercise also affects the GI. The objective of this study was, therefore, to determine the GI of common meals (three breakfast cereals: B, C, D) in healthy, nonsmoking young males 2 who were either endurance-trained (n = 12) or sedentary (n = 11). Glucose was used as the reference food. The GI value between the endurance-trained and sedentary subjects differed significantly (p < 0.01). Pair-wise comparisons between endurance-trained and sedentary subjects within the different test meals were significant for test meal D (p = 0.002), marginally non-significant for meal C (p = 0.052) and not significant for meal B (p = 0.204). These results suggest that the GI of some complex foods may depend on the training status of healthy young subjects. PMID- 16711657 TI - Unilateral intraputaminal glial cell line-derived neurotrophic factor in patients with Parkinson disease: response to 1 year each of treatment and withdrawal. AB - OBJECT: Glial cell line-derived neurotrophic factor (GDNF) infused unilaterally into the putamen for 6 months was previously shown to improve motor functions and quality of life measures significantly in 10 patients with Parkinson disease (PD) in a Phase I trial. In this study the authors report the safety and efficacy of continuous treatment for 1 year or more. After the trial was halted by the sponsor, the patients were monitored for an additional year to evaluate the effects of drug withdrawal. METHODS: During the extended study, patients received unilateral intraputaminal infusion of 30 mg/day GDNF at a basal infusion rate supplemented with pulsed boluses every 6 hours at a convection-enhanced delivery rate to increase tissue penetration of the protein. When the study was stopped, the delivery system was reprogrammed to deliver sterile saline at the basal infusion rate of 2 ml/hour. The Unified PD Rating Scale (UPDRS) total scores after 1 year of therapy were improved by 42 and 38%, respectively, in the "off" and "on" states. Motor UPDRS scores were also improved: 45 and 39% in the off and on conditions, respectively. Benefits from treatment were lost by 9 to 12 months after GDNF infusion was halted. At that time, the patients had returned to their baseline UPDRS scores and required higher levels of conventional antiparkinsonian drugs to treat symptoms. After 11 months of treatment, the delivery system had to be removed in one patient because of the risk of infection. In seven patients antibodies to GDNF developed, with no evidence of clinical sequelae. There was also no evidence of GDNF-induced cerebellar toxicity, as evaluated using magnetic resonance imaging analysis and clinical testing. CONCLUSIONS: Unilateral administration of GDNF results in significant, sustained bilateral benefits. These improvements are lost within 9 months after drug withdrawal. Safety concerns with GDNF therapy can be closely monitored and managed. PMID- 16711658 TI - Olivecrona on the development of neurosurgery in the middle of the twentieth century: reflections with the wisdom of today. AB - Herbert Olivecrona was one of the pioneers of modern neurosurgery. In one of his articles, published more than half a century ago, Olivecrona shared his thoughts on the development of neurosurgery in the first half of the twentieth century and his vision regarding the future of neurosurgery and neurosurgical training. In the present paper, the authors communicate their reflections and thoughts on Herbert Olivecrona's visionary article. PMID- 16711659 TI - The BION devices: injectable interfaces with peripheral nerves and muscles. AB - OBJECT: The purpose of this study was to describe a novel technology for implantable neuromuscular stimulation to treat complications of paralysis and disuse atrophy, including shoulder subluxation, hand contractures, drop foot, and osteoarthritis. The authors review the results so far of several pilot clinical studies of these muscle stimulation devices. METHODS: Miniature wireless stimulators received power and individually addressed command signals from an external radiofrequency transmission coil. One or more implants were injected through a 12-gauge hypodermic insertion tool into muscles or adjacent to motor nerves, where they provided the means to activate the muscles in any desired pattern of intensity and frequency. Randomized controlled studies in small numbers of patients are underway to identify efficacy, acceptability, best methods of practice, and any design changes that may be required to improve the technology. Fifty patients have been enrolled in five studies; 35 patients have undergone implantation of a total of 79 BION1 devices. Comparisons with surface stimulation in patients who have suffered a stroke with shoulder subluxation and hand contractures show similar improvements in objective measures of efficacy but higher comfort levels for stimulation by implants. CONCLUSIONS: Injected microstimulators represent a promising new class of technology for the rehabilitation of patients with upper motor neuropathies. As the technology evolves, practitioners may be able to use it to facilitate functional reanimation of paralyzed limbs. PMID- 16711660 TI - On the design of robotic hands for brain-machine interface. AB - Brain-machine interface (BMI) is the latest solution to a lack of control for paralyzed or prosthetic limbs. In this paper the authors focus on the design of anatomical robotic hands that use BMI as a critical intervention in restorative neurosurgery and they justify the requirement for lower-level neuromusculoskeletal details (relating to biomechanics, muscles, peripheral nerves, and some aspects of the spinal cord) in both mechanical and control systems. A person uses his or her hands for intimate contact and dexterous interactions with objects that require the user to control not only the finger endpoint locations but also the forces and the stiffness of the fingers. To recreate all of these human properties in a robotic hand, the most direct and perhaps the optimal approach is to duplicate the anatomical musculoskeletal structure. When a prosthetic hand is anatomically correct, the input to the device can come from the same neural signals that used to arrive at the muscles in the original hand. The more similar the mechanical structure of a prosthetic hand is to a human hand, the less learning time is required for the user to recreate dexterous behavior. In addition, removing some of the nonlinearity from the relationship between the cortical signals and the finger movements into the peripheral controls and hardware vastly simplifies the needed BMI algorithms. (Nonlinearity refers to a system of equations in which effects are not proportional to their causes. Such a system could be difficult or impossible to model.) Finally, if a prosthetic hand can be built so that it is anatomically correct, subcomponents could be integrated back into remaining portions of the user's hand at any transitional locations. In the near future, anatomically correct prosthetic hands could be used in restorative neurosurgery to satisfy the user's needs for both aesthetics and ease of control while also providing the highest possible degree of dexterity. PMID- 16711661 TI - Acute microelectrode array implantation into human neocortex: preliminary technique and histological considerations. AB - OBJECT: Researchers at The Center for Neural Interfaces at the University of Utah have designed and produced a silicon-based high-density microelectrode array that has been used successfully in mammalian models. The authors investigate the ability to transfer array insertion techniques to humans and examine the acute response of human cortical tissue to array implantation. METHODS: Six patients who were scheduled to undergo temporal lobectomy surgery were enrolled in an Institutional Review Board-approved protocol. Before the patients underwent lateral temporal cortical resection, one or two high-density microelectrode arrays were implanted in each individual by using a pneumatic insertion device. Cortical tissue was then excised and preserved in formalin. The specimens were sectioned and stained for histological examination. Pneumatic insertion of a microelectrode array into human cortex in the operating room was feasible. There were no clinical complications associated with implantation and no evidence of significant insertion-related hemorrhage. Tissue responses ranged from mild cortical deformity to small focal hemorrhages several millimeters below the electrode tines. Based on initial results, the insertion device was modified. A footplate that mechanically isolates a small area of cortex and a calibrated micromanipulator were added to improve the reproducibility of insertion. CONCLUSIONS: A high-density microelectrode array designed to function as a direct cortical interface device can be implanted into human cortical tissue without acute clinical complications. Further modifications to the insertion device and array design are ongoing and future work will assess the functional significance of the tissue reactions observed. PMID- 16711662 TI - Platform technologies to support brain-computer interfaces. AB - There is a lack of adequate and cost-effective treatment options for many neurodegenerative diseases. The number of affected patients is in the millions, and this number will only increase as the population ages. The developing areas of neuromimetics and stimulative implants provide hope for treatment, as evidenced by the currently available, but limited, implants. New technologies are emerging that are leading to the development of highly intelligent, implantable sensors, activators, and mobile robots that will provide in vivo diagnosis, therapeutic interventions, and functional replacement. Two key platform technologies that are required to facilitate the development of these neuromimetic and stimulative implants are data communication channels and the devices' power supplies. In the research reported in this paper, investigators have examined the use of novel concepts that address these two needs. These concepts are based on ionic volume conduction (VC) to provide a natural communication channel to support the functioning of these devices, and on biofuel cells to provide a continuously rechargeable power supply that obtains electrons from the natural metabolic pathways. The fundamental principles of the VC communication channels, including novel antenna design, are demonstrated. These principles include the basic mechanisms, device sensitivity, bidirectionality of communication, and signal recovery. The demonstrations are conducted using mathematical and finite element analysis, physical experiments, and animal experiments. The fundamental concepts of the biofuel cells are presented, and three versions of the cells that have been studied are discussed, including bacteria-based cells and two white cell-based experiments. In this paper the authors summarize the proof or principal experiments for both a biomimetic data channel communication method and a biofuel cell approach, which promise to provide innovative platform technologies to support complex devices that will be ready for implantation in the human nervous system in the next decade. PMID- 16711663 TI - Limits of brain-computer interface. Case report. AB - Most patients who are candidates for brain-computer interface studies have an injury to their central nervous system and therefore may not be ideal for rigorous testing of the full abilities and limits of the interface. This is a report on a quadriplegic patient who appeared to be a reasonable candidate for intracranial implantation of neurotrophic electrodes. He had significant cortical atrophy in both the motor and parietal cortical areas but was able to generate signal changes on functional magnetic resonance images by thinking about hand movements. Only a few low-amplitude action potentials were obtained, however, and he was unable to achieve single-unit control. Despite this failure, the use of field potentials offered an alternative method of control and allowed him some limited computer interactions. There are clearly limits to what can be achieved with brain-computer interfaces, and the presence of cortical atrophy should serve as a warning for future investigators that less invasive techniques may be a more prudent approach for this type of patient. PMID- 16711664 TI - Spinal arteriovenous malformations: new classification and surgical treatment. AB - OBJECT: Spinal vascular malformations represent rare and insufficiently studied pathological entities characterized by considerable variation. Insufficient study of this disease is connected with the complexity of its diagnosis, which restricts the development of surgical treatments that are differentiated according to the type of malformation. Great difficulties are caused by the lack of a clear structural-hemodynamic classification of spinal arteriovenous malformations (AVMs). At present the classification created between 1991 and 1998 by the combined efforts of different authors is the most widely used one. According to this classification, four categories are distinguishable: Type I, dural arteriovenous fistulas (AVFs); Type II, intramedullary glomus AVMs; Type III, juvenile or combined AVMs; and Type IV, intradural perimedullary AVFs. Vascular tumors are also classified, as follows: hemangiomas, hemangioblastomas, angiosarcomas, hemangiopericytomas, angiofibromas, angiolipomas, and hemangioendotheliomas, as well as cavernous malformations. METHODS: In this study the authors analyze the diagnostic data and results of treatment in 91 patients with AVMs and AVFs who were treated at the Institute of Neurosurgery between 1995 and 2005. The patients' ages ranged from 9 to 83 years; the mean age was 42.9 years. For spinal vascular malformations we devised a classification that took into account the aforementioned features of AVMs: the anatomical characteristics of a malformation and its angiostructural and hemodynamic features. In all patients the neuroimaging modalities used in the investigation of their lesions included magnetic resonance (MR) imaging and selective spinal angiography. Three dimensional computerized tomography angiography studies were obtained in 14 patients, and MR angiography was used in 17. CONCLUSIONS: For successful surgical treatment of spinal AVMs it is necessary to obtain data about their localization, vascular structure, and hemodynamics that are as complete as possible. This information will promote the use of optimum surgical procedures and the latest methods of microsurgical and endovascular interventions, with treatments differentiated according to the type of malformation. One should try to use the least invasive endovascular approach in these cases,where possible, to occlude the AVM or reduce the intensity of blood flow by means of embolization. To perform an AVM resection or occlusion, one should use a direct approach to the malformation, blocking only the vessels supplying blood to the malformation and preserving the vessels feeding the spinal cord. PMID- 16711665 TI - A technique for minimally altering anatomically based subthalamic electrode targeting by microelectrode recording. AB - OBJECT: Implantation of a subthalamic nucleus (STN) deep brain stimulation (DBS) electrode is increasingly recognized as an effective treatment for advanced Parkinson disease (PD). Despite widespread use of microelectrode recording (MER) to delineate the boundaries of the STN prior to stimulator implantation, it remains unclear to what extent MER improves the clinical efficacy of this procedure. In this report, the authors analyze a series of patients who were treated at one surgical center to determine to what degree final electrode placement was altered, based on readings obtained with MER, from the calculated anatomical target. METHODS: Subthalamic DBS devices were placed bilaterally in nine patients with advanced PD. Frame-based volumetric magnetic resonance images were acquired and then transferred to a stereotactic workstation to determine the anterior and posterior commissure coordinates and plane. The initial anatomical target was 4 mm anterior, 4 mm deep, and 12 mm lateral to the midcommissural point. The MERs defined the STN boundaries along one or more parallel tracks, refining the final electrode placement by comparison of results with illustrations in a stereotactic atlas. In eight of 18 electrodes, the MER results did not prompt an alteration in the anatomically derived target. In another eight placements, MER altered the target by less than 1 mm and two of 18 electrode positions differed by less than 2 mm. The anterior-posterior difference was 0.53 +/- 0.65 mm, whereas the medial-lateral direction differed by 0.25 +/- 0.43 mm. The ventral boundary of the STN defined by MER was 2 +/- 0.72 mm below the calculated target (all values are the means +/- standard deviation). All patients attained clinical improvement, similar to previous reports. CONCLUSIONS: In this series of patients, microelectrode mapping of the STN altered the anatomically based target only slightly. Because it is not clear whether such minor adjustments improve clinical efficacy, a prospective clinical comparison of MER refined and anatomical targeting may be warranted. PMID- 16711669 TI - Platinum(II) diimine diacetylides: metallacyclization enhances photophysical properties. AB - The synthesis, structural characterization, and photoluminescence properties of a new platinum(II) diimine complex bearing the bidentate diacetylide ligand tolan 2,2'-diacetylide (tda), Pt(dbbpy)(tda) [dbbpy = 4,4'-di-tert-butyl-2,2' bipyridine], are described. In CH2Cl2, Pt(dbbpy)(tda) exhibits a strong visible charge-transfer absorption and broad emission centered at 562 nm. The photoluminescence quantum yield and excited-state lifetime are 0.52 and 2.56 mus, respectively, at room temperature. These parameters indicate that the planarization and rigidity introduced by the cyclic diacetylide leads to a lower energy-absorbing species displaying enhanced photophysics relative to the analogous Pt(dbbpy)(CCPh)2. Time-dependent density functional theory calculations, which include solvation by CH2Cl2 via the polarizable continuum model, are used to reveal the nature of the excited states in these molecules that are responsible for the charge-transfer transitions. The 77 K emission spectra of the two compounds in EtOH/MeOH glasses are compared, uncovering tda based ligand-localized phosphorescence in the title compound. PMID- 16711666 TI - Potentiation of neurite outgrowth and reduction of apoptosis by immunosuppressive agents: implications for neuronal injury and transplantation. AB - OBJECT: Immunosuppressive agents are believed to play a role in recovery from spinal cord injury, but the underlying mechanisms by which neuronal function is improved by these agents are poorly understood. In this study, the authors evaluate the effect of immunosuppressive medications on neurite outgrowth and cell survival after a pharmacologically induced injury. METHODS: Differentiated human neuroblastoma SH-SY5Y cells were injured using the calcium agonist thapsigargin. After cellular injury, neurite outgrowth in the presence or absence of immunosuppressive agents was measured. Apoptosis was quantified with the aid of a terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling assay. Neurite outgrowth was severely restricted following thapsigargin injury. Outgrowth was potentiated, however, by the addition of concentrations of 1 and 10 mM cyclosporin A in a dose-dependent fashion. Similarly, addition of 10 nM FK506 increased the percentage of neurites in the 20 to 40-micron range. A low dose (1 mM) of dexamethasone did not have a significant effect on neurite outgrowth, but a higher dose (10 mM) increased the percentage of neurites in the 10- to 45-micron range. These agents also lessened the degree of thapsigargin-induced apoptosis. CONCLUSIONS: Immunosuppressive agents such as cyclosporin A, FK506, and dexamethasone can potentiate neurite outgrowth and protect against apoptotic cell death in a human postmitotic neuronal cell line. Such effects may have implications for lessening neuronal injury after neurotrauma, stroke, or neurodegeneration. PMID- 16711670 TI - A two-dimensional octacyanomolybdate(V)-based ferrimagnet: {[Mn(II)(DMF)4]3[Mo(V)(CN)8]2}n. AB - Treatment of [HNBu3]3[Mo(V)(CN)8] with manganese(II) p-toluenesulfonate in N,N' dimethylformamide (DMF) affords {[Mn(II)(DMF)4]3[Mo(V)(CN)8]2}n (1) as a two dimensional network. The structure of 1 consists of [cis-Mn(II)(DMF)4(mu-NC)2]2+ and [trans-Mn(II)(DMF)4(mu-NC)2]2+ units that are linked via cyanides to three connected [Mo(V)(CN)5(mu-CN)3]3- centers in a 4:2:6 ratio, forming 12-membered rings. Magnetic measurements indicate that 1 is a ferrimagnet (TN = 8 K) that exhibits frequency-dependent behavior in chi". Heating of 1 affords an additional magnetic phase (TN = 21 K) that is absent of linkage isomerism. PMID- 16711671 TI - Supramolecular framework with two different kinds of channels via self-assembly of 1D coordination polymers in a prismatic manner. AB - Slow diffusion of Cu(ClO4)2 with p-C6H4(SiMe2(4-Py))2 yields a supramolecular framework consisting of two kinds of channels via self-assembly of a 1D double stranded chain in a prismatic fashion. The channel has a 11 x 11 A2 cross section (Cu...C = 11.13 A) with a 6 x 6 A2 square pore. The channels are infinitely arranged, resulting in another kind of channel with similar pores. Thus, two different kinds of channels with different solvate molecules coexist in a 1:2 ratio. Weak C-H...pi interaction may be one of the driving forces in the assembly of the prismatic channel structure. PMID- 16711672 TI - Synthesis, characterization, and crystal structures of two novel high-nuclear nickel-substituted dimeric polyoxometalates. AB - Two novel high-nuclear Ni-substituted polyoxometalates Na9K[Ni7(OH)4(H2O)(CO3)2(HCO3)(A-alpha-SiW9O34)(beta-SiW10O37)].5H3O.18H2O (1) and K6Na4[{Ni6(H2O)4(mu2-H2O)4(mu3-OH)2}(x-SiW9O34)2].17.5H2O (2) have been obtained by reaction of K(8)[gamma-SiW10O36] with NiCl2.2H2O in basic conditions. Compounds 1 and 2 all represent the first polyoxometalates substituted by seven and six Ni centers, respectively. Also, compound 1 exhibits a novel asymmetric dimeric structure, composed of [A-alpha-SiW9O34]10- and [beta-SiW10O37]10- units linked by a Ni7 cluster and three carbonates, which exhibit tri- and bidentate coordination modes in 1. PMID- 16711673 TI - Unexpected photoisomerization of a pincer-type amido ligand leads to facial coordination at Pt(IV). AB - The divalent complex (BQA)PtMe undergoes oxidative addition with MeI to afford the octahedral complex cis-(mer-BQA)PtMe2I {(BQA)- = bis(8-quinolinyl)amide}. When this molecule is irradiated with visible light, it isomerizes to (fac BQA)PtMe2I, where the BQA ligand adopts an unexpected facial coordination mode. The amide nitrogen in this molecule is sp3 hybridized and can be easily quarternized with HBF4, resulting in [H(fac-BQA)PtMe2I][BF4], with only minor perturbation to the coordination sphere. PMID- 16711674 TI - Photoinduced electron-transfer reactions of platinum(II) terpyridyl acetylide complexes: reductive quenching in a hydrogen-generating system. AB - The one-electron reduction of [(mpt)Pt(CCPhCl)] ClO4 [mpt = 4'-(4-methylphenyl) 2,2',6',2' '-terpyridyl; CCPhCl = chlorophenyl acetylide] by a series of amine donors was investigated in a CH3CN solution. The excited state has a lifetime of 550 ns in the absence of quencher in degassed CH3CN. Quenching rate constants were >10(8) M(-1) s(-1) even for trialkylamines. Transient absorption spectroscopy provided convincing evidence for the production of the one-electron reduced Pt complex, even with quenchers that could potentially serve as H-atom donors. The transient spectrum obtained in the presence of triethylamine exhibits, in addition to bleaching of the Pt(II) complex absorption, a maximum at approximately 360 nm and broad absorption from 500 to 800 nm, indicating that the reduced complex has a complex electronic structure and is not easily assigned as a terpyridyl anion radical. PMID- 16711675 TI - Stepwise guest adsorption with large hysteresis in a coordination polymer {[Cu(bhnq)(THF)2](THF)}n constructed from a flexible hingelike ligand. AB - Reversible vapochromic behavior of a porous copper(II) coordination polymer {[Cu(bhnq)(THF)2](THF)}n (1; THF = tetrahydrofuran) constructed from a flexible hingelike ligand H2bhnq [2,2'-bis(3-hydroxy-1,4-naphthoquinone)] has been investigated by adsorption measurements. The isotherms show large hysteretic and stepwise profiles, suggesting the occurrence of the guest-induced framework transformation. The dynamic coil-like behavior of 1 can be controlled through the change of the hydrogen-bonding interactions caused by the reversible and selective incorporation of guest molecules. PMID- 16711676 TI - Structure and magnetic properties of a hydroxo-bridged copper(II) distorted cubane stabilized via supramolecular hydrogen bonding with an ionic hexafluoroacetylacetonate. AB - Tetranuclear [Cu(II)4(OH)4(aib)4)](hfac)4 (1; aib = 2-methyl-2-amino-4 iminopentane; hfac = hexafluoroacetylacetonate) forms from the reaction of aqueous ammonia and Cu(hfac)2.2H2O in acetone. The structure of 1 reveals that four noncoordinating hfac- counterions stabilize the distorted cubane complex via multiple H-bonding contacts. Magnetic susceptibility studies reveal that cubane like 1 is best described as a pair of independent antiferromagnetically coupled dimers with g = 2.10 and J/kB = -298 K (207 cm(-1)) (H = -2JS1.S2). PMID- 16711677 TI - A diamagnetic dititanium(III) paddlewheel complex with no direct metal-metal bond. AB - Reaction of Ti[N(But)Ar]3 (Ar = 3,5-C6H3Me2 or Ar' = C6H5) with CO2 at -40 degrees C produces diamagmetic Ti(III) paddlewheel complexes with long Ti-Ti separations (>3.4 Angstrom), thus excluding direct Ti-Ti bonding. 1H NMR spectroscopy shows that the compounds are diamagnetic in solution in the temperature range of -65 to +70 degrees C. In the solid state, the diamagnetism was found to persist between 2 and 300 K. Calculations at the density functional theory level suggest that the diamagnetism results from antiferromagnetic coupling by superexchange through the ligand pi system. PMID- 16711678 TI - p-Carborane: a new cage spacer for photoactive metal polypyridine dyads. AB - The first example of a binuclear ruthenium complex involving the p-carborane framework in the bridging ligand is reported. The bridging ligand is a symmetric linear array comprising a central p-carborane unit, two p-phenylene spacers, and two 5-yl-2,2'-bipyridine coordinating units. A homobinuclear Ru(II) complex, with 2,2'-bipyridine as peripheral ligands, was synthesized and characterized. The Ru(II)-Ru(III) mixed-valence species, obtained by partial oxidation, has been investigated with steady-state and time-resolved techniques in CH3CN. The rate of photoinduced electron transfer is 2.3 x 10(8) s(-1). PMID- 16711679 TI - [Ni(xbsms)Ru(CO)2Cl2]: a bioinspired nickel-ruthenium functional model of [NiFe] hydrogenase. AB - As a model of the active site of [NiFe] hydrogenases, a dinuclear nickel ruthenium complex [Ni(xbsms)Ru(CO)2Cl2] was synthesized and fully characterized. The three-dimensional structure reveals a nickel center in a square-planar dithioether-dithiolate environment connected to a ruthenium moiety via a Ni(mu SR)2Ru bridge. This complex catalyzes hydrogen evolution by electroreduction of the weakly acidic Et3NH+ ions in N,N-dimethylformamide and is therefore the first functional bioinspired model of [NiFe] hydrogenases. PMID- 16711680 TI - New N,N,N-heteroscorpionates based on 2,2'-bis(pyrazolyl)ethanamine and its derivatives. Ligands designed for probing supramolecular interactions. AB - The successful design and synthesis of the new bis(pyrazolyl)ethanamine ligand and its copper(I) triphenylphosphine complex is reported. The ligand coordinates to the copper(I) center in a fac tridentate fashion, through both the pyrazolyl rings and the nitrogen atom from the NH2 group. In the solid state, the compound is organized in a 2D noncovalent network by N-H...pi and C-H...pi interactions and hydrogen bonds. The analogous ligand with a benzyl group substituted on the amine forms a complex with the same copper(I) center that has a similar 2D supramolecular structure and, in addition, is organized by the benzyl synthon into a 3D architecture. PMID- 16711681 TI - Multinuclear luminescent Schiff-base Zn-Nd sandwich complexes. AB - Multinuclear 3d-4f complexes with sandwichlike molecular structures are formed with the Schiff-base ligand bis(3-methoxysalicylidene)ethylene-1,2 phenylenediamine(H(2)L). The stoichiometry and structures are dependent on the Zn:Nd ratio and counteranions present. They are trinuclear [Nd(ZnL)2(NO3)2(H2O)2].NO3.EtOH.H2O (1), [Nd(ZnL)2Cl2(H2O)3].Cl.2MeOH.5H2O (2), and tetranuclear [Nd2(ZnL)2Cl6(MeOH)2].MeOH (3). Dinuclear complex [NdZnL(NO3)3MeCN].MeCN (4) was also characterized. Near-infrared (NIR) lanthanide luminescence is observed in these complexes. PMID- 16711682 TI - The room-temperature superstructure of ZrP2O7 is orthorhombic: there are no unusual 180 degrees P-O-P bond angles. AB - The structure of room-temperature ZrP2O7 is shown to be orthorhombic by a combination of high-resolution synchrotron powder diffraction and single-crystal synchrotron diffraction data. Small nontwinned single crystals were obtained by synthesizing the compound using solvothermal methods at temperatures below the cubic to orthorhombic phase transition. The average P-O-P angle is 146 degrees. DFT calculations (B3LYP/AUG-cc-pVDZ) on the isolated P2O7(4-) anion yield a P-O-P angle of 153.42 degrees and indicate that the barrier to inversion is of the order 3.6 kJ mol(-1). PMID- 16711683 TI - The 136-atom structure of ZrP2O7 and HfP2O7 from powder diffraction data. AB - There has been considerable debate in the literature about the true room temperature structure of ZrP2O7 and related materials. In this article we describe how a combination of information from solid-state 31P NMR and powder diffraction data can be used to determine the structure of this 136 unique-atom material. The structure has been solved using a combination of simulated annealing and Rietveld refinement performed simultaneously on X-ray and neutron diffraction data. Despite the close to cubic metric symmetry of the material, we show how its true orthorhombic structure (space group Pbca) can be refined to a high degree of precision. PMID- 16711684 TI - Structure of anhydrous alkali-metal formates. AB - Single-crystal X-ray diffraction has yielded new crystal structures for cesium formate (CsOOCH) and rubidium formate (RbOOCH). The cesium formate structure has the same unit cell and space group as that published from powder X-ray diffraction data but differs radically in the placement and orientation of the formate ions. The new crystal structure has been successfully modeled with an empirical force field based on pair potentials, whereas it proved impossible to develop a force field that gave an adequate description of the powder structure. For rubidium formate, the gross structure is similar to that previously published, but the space group includes a mirror plane (Pbcm rather than Pbc2(1)). From this information, we have been able to analyze the effect of the cation size on the crystal structure for alkali-metal formates. PMID- 16711685 TI - Intercalation and photophysical properties of the tetra-(8-hydroxyquinolinato) boron complex and 3,3',4,4'-benzophenone tetracarboxylic anion into Mg-Al layered double hydroxides. AB - The tetra-(8-hydroxyquinolinato) boron complex (Bq) and 3,3',4,4'-benzophenone tetracarboxylic anion (BPTC) were successfully intercalated into layered double hydroxides with an ion-exchange method. Elemental analyses, powder X-ray diffraction, FTIR spectra, and thermogravimetric analyses indicated that the hydrothermal treatment could efficiently improve the reaction process, the purity and crystallinity of products, and that Bq and BPTC molecules steadily arranged with C2 axis perpendicular to the layer plane, and the J-type aggregate in the host interlayer, respectively. The fluorescence studies showed that, compared with those observed in the dilute solutions of the guests, the spectra of Mg3Al Bq and Mg3Al-BPTC evolved to the high-energy side and the low-energy side with increasing their loading, respectively, which were attributed to various arrangements of guests and the increased intermolecular interaction. Furthermore, their fluorescence intensity gradually decreased with increasing the intercalated guest content due to the concentration quenching, and Mg3Al-Bq exhibits enhanced solid-state blue luminescence due to a more rigid and constrained environment of the host. PMID- 16711686 TI - Synthesis and characterization of a tripodal amide ligand and its binding with anions of different dimensionality. AB - Synthesis and crystal structure of a tren-based amide, L(1), N,N',N''-tris[(2 amino-ethyl)-3-nitro-benzamide] is reported. The crystallographic results show intramolecular hydrogen bonding and aromatic pi...pi stacking among tripodal arms which prevent opening of the receptor cavity. Intermolecular hydrogen bonding in L1 generates the sheetlike network in the solid state. The structural aspects of binding halides (1 and 2), nitrate (3), perchlorate (4), and hexafluorosilicate (5) with the protonated L1 are examined crystallographically. Protonation at the apical nitrogen of L1 in the presence of anions shows a structural transformation from sheet to bilayer network. Anion binding with multiple receptor units is observed via amide N-H...anion and aryl C-H...anion hydrogen-bonding interactions in all the complexes. The aryl group having nitro functionality that contributes to anion binding in complexes 1-5 through CH...anion interactions (either para or meta to nitro C-H) is noteworthy. These studies also show higher anion coordination of chloride (8) and hexafluorosilicate (14) with L1H+. PMID- 16711687 TI - From an S(T) = 3 single-molecule magnet to diamagnetic ground state depending on the molecular packing of Mn(III)salen-type dimers decorated by N,N'-dicyano-1,4 naphthoquinonediiminate radicals. AB - Two manganese(III) tetradentate Schiff-base dimers to which N,N'-dicyano-1,4 naphthoquinonediiminate (DCNNQI) radicals are attached have been selectively synthesized by varying the solvents used in the reactions: [Mn2(5 MeOsaltmen)2(DCNNQI)2].MeOH (1) and [Mn2(5-MeOsaltmen)(2)(DCNNQI)(2)] x 2CH2Cl2.2CH3CN (2) [5-MeOsaltmen2- = N,N'-(1,1,2,2-tetramethylethylene)bis(5 methoxysalicylideneiminate)]. These two complexes share the same molecular core, [(DCNNQI.-)-Mn(III)-(O)2-Mn(III)-(DCNNQI.-)], where -(O)2- is a biphenolate bridge in the out-of-plane dimerized [Mn(2)(5-MeOsaltmen)2]2+ moiety. However, their packing arrangements are completely different. Whereas complex 1 is found to be relatively isolated, strong intermolecular dimerization of the DCNNQI moieties (with the nearest contact being approximately 3.0 A) is observed in 2, forming a one-dimensional chain of [-Mn(III)-(O)2-Mn(III)-(DCNNQI.-)2 ](infinity). The magnetic susceptibility of 1 can be modeled with an [S = 1/2, 2, 2, 1/2] four-spin system including strong antiferromagnetic Mn(III)/DCNNQI radical coupling (J(Mn/rad)/kB = -23 K) and ferromagnetic Mn(III)/Mn(III) coupling through the biphenolate bridge (J(Mn/Mn)/kB = +2.0 K). These interactions lead to an ST = 3 ground state that possesses significant uniaxial anisotropy (D(S=3)/kB = -2.1 K). Low-temperature ac and dc magnetic data of 1 reveal its single-molecule magnet behavior with quantum tunneling of the magnetization. By contrast, 2 possesses the diamagnetic ground state induced by dominating Mn(III)-Mn(III) antiferromagnetic interactions mediated by the diamagnetic DCNNQI dimers and/or pi-pi contact along the b axis. PMID- 16711688 TI - Single-crystal structure of the 2H-related perovskites (A(3-x)Nax)NaBO6 (A = La, Pr, Nd; B = Rh, Pt). AB - Single crystals of La(2.47)Na(1.53)RhO6, Pr(2.45)Na(1.55)RhO6, Nd(2.45)Na(1.55)RhO6, La2Na2PtO6, and Nd2Na2PtO6 were grown from carbonate and "wet" hydroxide fluxes. All were found to crystallize in the trigonal space group R3c and adopt the K4CdCl6 structure. PMID- 16711689 TI - Metal-metal bonding in mixed valence Ni2(5+) complexes and spectroscopic evidence for a Ni2(6+) species. AB - Dinickel(II) complexes of the ligands N,N'-di-p-anisylformamidinate (DAniF) and N,N',N''-triphenylguanidinate (TPG) have been synthesized and crystallographically characterized, along with their one-electron-oxidized analogues. In both systems, the Ni-Ni distances become shorter by approximately 0.1 A upon oxidation, in accord with the proposal that the resulting Ni2(5+) complexes are appropriately described as having one electron removed from a metal based sigma orbital and an overall Ni-Ni bond order of 1/2. Although conventional DFT calculations on the model compounds Ni2(HNCHNH)4 and [Ni2(HNCHNH)4]+ appear to predict that the lowest energy state of the latter species would have one unpaired electron in an essentially ligand-based orbital. A single-point calculation of Ni2(DAniF)4 employing the geometry of its crystal structure with the full ligand included reveals a reversal of the previously predicted order of the HOMO and HOMO-1, and suggests that the unpaired electron in [Ni2(DAniF)4]+ is in a metal-based orbital of sigma symmetry. This is verified by the axial EPR spectrum of the compound in solution. The compound Ni2(DAniF)4 shows an unexpectedly rich cyclic voltammogram with four stepwise reversible oxidation waves. Coulometric experiments show that the doubly oxidized species has a significant lifetime at -25 degrees C, and by spectroelectrochemistry, its UV-vis spectrum was recorded. We propose that this species contains a Ni2(6+) core with a single Ni-Ni sigma bond. PMID- 16711690 TI - Molecular precursor route to bulk and silica-supported Nb2Mo3O14 using water soluble oxo-oxalato complexes. AB - The catalytically relevant Nb2Mo3O14 phase has been prepared in bulk and silica supported forms via the so-called "multiple molecular precursors method" from water-soluble oxo-oxalato complexes of Nb and Mo, (NH4)3[NbO(ox)3].H2O, and (NH4)2[MoO3(ox)].H2O. Thermal treatment of the mixed Nb-Mo precursor has been optimized for the formation of the pure Nb2Mo3O14 phase, either as bulk oxide or a silica-supported phase with high specific surface area. A characterization of the bulk phase obtained via the conventional ceramic route has also been carried out and a comparison has been made with the precursors route. According to this route, the Nb2Mo3O14 phase is shown to be formed in a pure form at 700 degrees C (i.e., 100 degrees C below the lowest temperature reported so far for the formation of the phase by the ceramic method). The supported samples have appreciable specific surface areas of 60-70 m(2) g(-1), much larger than those reached in the previous attempts under vacuum in sealed vials. The SEM and EDX analyses reveal a high dispersion of the desired phase on the silica support. PMID- 16711691 TI - Influence of the counterion and the solvent molecules in the spin crossover system [Co(4-terpyridone)2]Xp.nH2O. AB - A series of new complexes belonging to the [Co(4-terpyridone)2]X(p) x nS family (4-terpyridone = 2,6-bis(2-pyridyl)-4(1H)-pyridone) have been synthesized and characterized, using X-ray single crystal determination and magnetic susceptibility studies, to be X = [BF4]- (p = 2) and S = H2O for polymorphs 1 and 2, X = [BF4]- (p = 1) and [SiF6]2- (p = 0.5) and S = CH(3)OH for 3, X = [SiF6]2- (p = 1) and S = 3CH3OH and H2O for 4, X = [Co(NCS)4]2- (p = 1) and S = 0.5CH3OH for 5, X = I- (p = 2) and S = 5H2O for 6, X = [PF6]- (p = 1) for 7, and X = [NO3] (p = 2) for 8. Compounds 1-7 can be grouped in three sets according to the space group in which they crystallize: (i) P1 triclinic (1, 3), (ii) P2(1) monoclinic (2), and (iii) P2(1)/c monoclinic (4-7). The tridentate 4-terpyridone ligands coordinate the Co(II) ions in a mer fashion defining essentially tetragonally compressed [CoN6] octahedrons. The Co-N axial bonds involving the pyridone rings are markedly shorter than the Co-N equatorial bonds collectively denoted as Co N(central) and Co-N(distal), respectively. The differences in the average Co N(central) or Co-N(distal) distances observed for 1-7 reflect the different spin states of Co(II). Complexes 7 and 4' are fully high spin (HS), while 5 and 6 are low spin (LS). However, the counterion [Co(NCS)4]2- in complex 5 is high spin. Complexes 1, 2, 3, and 8 exhibit spin-crossover behavior in the 400-100 K temperature region. Compounds 1 and 2 are polymorphs, and interestingly, 1 irreversibly transforms into 2 above 340 K because of a crystallographic phase transition which involves a drastic modification of the crystal packing. The relevant thermodynamic parameters associated with the spin transition of polymorph 2 have been estimated using the regular solution theory leading to DeltaH = 3.04 kJ mol(-1), DeltaS = 20 J K(-1) mol(-1), and Gamma = 0.95 kJ mol( 1). PMID- 16711692 TI - Self-assembly, structures, and solution dynamics of emissive silver metallacycles and helices. AB - Reactions of 9,10-bis(diphenylphosphino)anthracene (PAnP) and AgX (X = OTf-, ClO4 , PF6-, and BF4-) led to luminescent Ag-PAnP complexes with rich structural diversity. Helical polymers [Ag(mu-PAnP)(CH3CN)X]n (X = OTf-, ClO4-, and PF6-) and discrete binuclear [Ag2(mu-PAnP)2(CH3CN)4](PF6)2, trinuclear [Ag3(mu-PAnP)3 supersetBF4](BF4)2, and tetranuclear [Ag4(mu-PAnP)4 superset(ClO4)2](ClO4)2 metallacycles were isolated from different solvents. The tri- and tetranuclear metallacycles exhibited novel puckered-ring and saddlelike structures. Variable temperature (VT) 31P{1H}-NMR spectroscopy of the complexes was solvent dependent. The dynamics in CD3CN involve two species, but the exchange processes in CD2Cl2 are more complicated. A ring-opening polymerization was proposed for the exchange mechanism in CD3CN. PMID- 16711693 TI - Homochiral porous lanthanide phosphonates with 1D triple-strand helical chains: synthesis, photoluminescence, and adsorption properties. AB - Four homochiral porous lanthanide phosphonates, [Ln(H2L)3].2H2O, (H3L = (S) HO3PCH2-NHC4H7-CO2H, Ln = Tb (1), Dy (2), Eu (3), Gd (4)), have been synthesized under hydrothermal conditions. These compounds are isostructural, and they possess a 3D supramolecular framework built up from 1D triple-strand helical chains. Each of the helical chain consists of phosphonate groups bridging adjacent Ln(III) ions. The helical chains are stacked through hydrogen bonds to form 1D tubular channels along the c axis. Moreover, helical water chains are located in the 1D channels, and after removal of these water chains, the compounds exhibit selective adsorption capacities for N2, H2O, and CH3OH molecules. Compounds 1 and 3 show strong green and red fluorescent emissions, respectively, in the solid state at room temperature. Crystal data for 1: TbP3O17N3C18H37, tetragonal (No.76), space group P4(1), a = 12.4643(3) Angstrom, b = 12.4643(3) Angstrom, c = 18.7577(5) Angstrom, V = 2914.17(13) Angstrom(3), and Z = 4. For 2: DyP3O17N3C18H37, a = 12.4486(3) Angstrom, b = 12.4486(3) Angstrom, c = 18.7626(5) Angstrom, V = 2907.60(13) Angstrom(3), and Z = 4. For 3, EuP3O17N3C18H37, a = 12.4799(3) Angstrom, b = 12.4799(3) Angstrom, c = 18.8239(5) Angstrom, V = 2931.78(13) Angstrom(3), and Z = 4. For 4: GdP3O17N3C18H37, a = 12.4877(18) Angstrom, b = 12.4877(18) Angstrom, c = 18.824(4) Angstrom, V = 2935.5(8) Angstrom(3), and Z = 4. PMID- 16711694 TI - Importance of the O-M-O bridges (M = V5+, Mo6+) for the spin-exchange interactions in the magnetic oxides of Cu2+ ions bridged by MO4 tetrahedra: spin lattice models of Rb2Cu2(MoO4)3, BaCu2V2O8, and KBa3Ca4Cu3V7O28. AB - The spin-lattice models relevant for the magnetic oxides Rb2Cu2(MoO4)3, BaCu2V2O8, and KBa3Ca4Cu3V7O28 were determined by evaluating the relative strengths of the spin-exchange interactions between their Cu2+ ions on the basis of spin dimer analysis. Our study shows that the O-M-O bridges (M = V5+, Mo6+) between the magnetic ions Cu2+, provided by the MO4 tetrahedra, are crucial for the spin-exchange interactions and hence for deducing the spin-lattice models needed to interpret the magnetic properties of these oxides. The spin-lattice model of Rb2Cu2(MoO4)3 is not a uniform chain but two interpenetrating spin ladders that interact weakly with geometric spin frustration. The spin-lattice model of BaCu2V2O8 is an alternating chain as expected, but the spin-exchange paths responsible for it differ from those expected. With respect to the strongest spin exchange of BaCu2V2O8, the spin exchange of KBa3Ca4Cu3V7O28 is only slightly weaker, but the strongest spin exchange of Rb2Cu2(MoO4)3 is much weaker. This difference in the spin-exchange strengths is caused by the difference in the bridging modes of the MO4 tetrahedra leading to these spin exchange interactions. PMID- 16711695 TI - Iron porphyrin-cyclodextrin supramolecular complex as a functional model of myoglobin in aqueous solution. AB - The 1:1 inclusion complex of 5,10,15,20-tetrakis(4-sulfonatophenyl)porphinato iron(II) (Fe(II)TPPS) and an O-methylated beta-cyclodextrin dimer having a pyridine linker (1) binds dioxygen reversibly in aqueous solution. The O2 adduct was very stable (t(1/2) = 30.1 h) at pH 7.0 and 25 degrees C. ESI-MS and NMR spectroscopic measurements and molecular mechanics (MM) calculations indicated the inclusion of the sulfonatophenyl groups at the 5- and 15-positions of Fe(III)TPPS or Fe(II)TPPS into two cyclodextrin moieties of 1 to form a supramolecular 1:1 complex (hemoCD1 for the Fe(II)TPPS complex), whose iron center is completely covered by two cyclodextrin moieties. Equilibrium measurements and laser flash photolysis provided the affinities ( and ) and rate constants for O2 and CO binding of hemoCD1 (k(O2)(on), k(O2)(off), k(CO)(on), and k(CO)(off)). The CO affinity relative to the O2 affinity of hemoCD1 was abnormally high. Although resonance Raman spectra suggested weak back-bonding of d(pi)(Fe) --> pi(CO) and hence a weak CO-Fe bond, the CO adduct of hemoCD1 was very stable. The hydrophobic CO molecule dissociated from CO-hemoCD1 hardly breaks free from a shallow cleft in hemoCD1 surrounded by an aqueous bulk phase leading to fast rebinding of CO to hemoCD1. Isothermal titration calorimetry furnished the association constant (K(O2)), DeltaH degrees , and DeltaS degrees for O2 association to be (2.71 +/- 0.51) x 10(4) M(-1), -65.2 +/- 4.4 kJ mol(-1), and -133.9 +/- 16.1 J mol(-1) K(-1), respectively. The autoxidation of oxy hemoCD1 was accelerated by H+ and OH-. The inorganic anions also accelerated the autoxidation of oxy-hemoCD1. The O2-Fe(II) bond is equivalent to the O2.--Fe(III) bond, which is attacked by the inorganic anions or the water molecule to produce met-hemoCD1 and a superoxide anion. PMID- 16711696 TI - Testing bridge-mediated differences in dinuclear valence tautomeric behavior. AB - Two structurally characterized dinuclear valence tautomers are described. Cobalt ions are bridged by p- and m-phenylene units connected to 2,2'-bipyridines. X-ray crystal structures show that the molecules are in the [(Co(III))(Co(III))] forms at ca. 125 K, while spectroscopic studies show that both molecules can achieve the [(Co(II))(Co(II))] form above 400 K and confirm the [(Co(III))(Co(III))] form below 10 K. Magnetic susceptibility studies are also included. Our results highlight the necessity of studying both crystalline and amorphous samples to distinguish the effects of intrinsic electronic structure and intermolecular forces on valence tautomeric behavior. PMID- 16711697 TI - Metal and ligand K-edge XAS of titanium-TEMPO complexes: determination of oxidation states and insights into Ti-O bond homolysis. AB - Ti-TEMPO (TEMPO = 2,2,6,6-tetramethylpiperidine-N-oxyl) provides a means for generating Ti(III) complexes by homolysis of the Ti-O bond. It has been determined that bis-Cp-Ti-TEMPO complexes readily undergo homolytic cleavage while the mono-Cp-Ti-TEMPO complexes do not. Here Ti K- and Cl K-edge XAS are applied to directly determine the oxidation state of TiCl3TEMPO, TiCpCl2TEMPO, and TiCp2ClTEMPO, with reference to Ti(III) and Ti(IV) complexes of known oxidation state. The Ti K-edge data show that Ti(III) complexes exhibit a pre edge feature approximately 1 eV lower than any of the Ti(IV) complexes; while the Cl K-edges show that Ti(III) complexes have a Cl K- pre-edge feature to approximately 1 eV higher energy than any of the Ti(IV) complexes. Taken together, the Ti and Cl K-edge data indicate that the Ti-TEMPO complexes are best described as Ti(IV)-TEMPO anions (rather than Ti(III)-nitroxyl radicals). In addition, the Cl K-edges indicate that replacement of Cl by Cp weakens the bonding with the remaining ligands, with the Cl 3p covalency decreasing from 25% to 21% to 17% on going from TiCl3TEMPO to TiCpCl2TEMPO to TiCp2ClTEMPO. DFT calculations also show that the electronic structures of the Ti-TEMPO complexes are modulated by the replacement of chloride by Cp. The effect of the Cp on the ancillary ligation is one factor that contributes to facile Ti-O bond homolysis in TiCp2ClTEMPO. However, the results indicate the primary contribution to the energetics of Ti-O bond homolysis in TiCp2ClTEMPO is stabilization of the three coordinate product by Cp. PMID- 16711698 TI - In search of benzene-like Sn6(6-): synthesis of Na4CaSn6 with interconnected cyclohexane-like Sn6(6-). AB - The title compound was synthesized in an attempt to produce stacked benzene-like Sn6(6-) rings separated by alkaline-earth cations in analogy with the recently reported stacks of aromatic cyclopentadienyl-like Sn5(6-) in Na8BaSn6 (in addition to isolated Sn4- anions). The resulting compound, synthesized from a stoichiometric mixture of the elements at high temperature, has the "correct" stoichiometry with six tin atoms and six positive charges. However, the rings of Sn6(6-) are puckered into chair-type cyclohexanes that are interconnected into isolated cylindrical tubes stuffed with Ca2+ between the rings. Such tubes, if fused to each other, form the hexagonal diamond structure. The new compound is electronically balanced according to magnetic and four-probe resistivity measurements. Reported are also the synthesis and properties of Na10EuSn12 and Na10YbSn12 which are isostructural with the known Na10CaSn12. PMID- 16711699 TI - Lanthanide(III) complexes with a tetrapyridine pendant-armed macrocyclic ligand: 1H NMR structural determination in solution, X-ray diffraction, and density functional theory calculations. AB - Complexes between the tetrapyridyl pendant-armed macrocyclic ligand (L) and the trivalent lanthanide ions have been synthesized, and structural studies have been made both in the solid state and in aqueous solution. The crystal structures of the La, Ce, Pr, Gd, Tb, Er, and Tm complexes have been determined by single crystal X-ray crystallography. In the solid state, all the cation complexes show a 10-coordinated geometry close to a distorted bicapped antiprism, with the pyridine pendants situated alternatively above and below the main plane of the macrocycle. The conformations of the two five-membered chelate rings present in the complexes change along the lanthanide series. The La(III) and Ce(III) complexes show a lambdadelta (or deltalambda) conformation, while the complexes of the heavier lanthanide ions present lambdalambda (or deltadelta) conformation. The cationic [Ln(L)]3+ complexes (Ln = La, Pr, Eu, Tb, and Tm) were also characterized by theoretical calculations at the density-functional theory (DFT) B3LYP level. The theoretical calculations predict a stabilization of the lambdalambda (or deltadelta) conformation on decreasing the ionic radius of the Ln(III) ion, in agreement with the experimental evidence. The solution structures show a good agreement with the calculated ones, as demonstrated by paramagnetic NMR measurements (lanthanide induced shifts and relaxation rate enhancements). The 1H NMR spectra indicate an effective D2 symmetry of the complexes in D2O solution. The 1H lanthanide induced shifts (LIS) observed for the Ce(III), Tm(III), and Yb(III) complexes can be fit to a theoretical model assuming that dipolar contributions are dominant for all protons. The resulting calculated values are consistent with highly rhombic magnetic susceptibility tensors with the magnetic axes being coincident with the symmetry axes of the molecule. In contrast with the solid-state structure, the analysis of the LIS data indicates that the Ce(III) complexes present a lambdalambda (or deltadelta) conformation in solution. PMID- 16711701 TI - Mass spectrometric and quantum chemical studies of the thermodynamics and bonding of neutral and ionized LnCl, LnCl2, and LnCl3 species (Ln = Ce, Lu). AB - The mass spectral patterns of CeCl3(g) and LuCl3(g) and appearance energies for the identified ions were measured using a Nier-type mass spectrometer coupled with a Knudsen cell. The molecular ion CeCl3+ was found to be considerably less stable in comparison to LuCl3+. Partial pressures and sublimation enthalpies of LnCl3(s) to monomeric LnCl3(g) and dimeric Ln2Cl6(g) species were obtained in the ranges of 882-1028 (Ln = Ce) and 850-1004 K (Ln = Lu). The contribution of dimeric Ce2Cl6(g) species to equilibrium vapors of CeCl3(s) is considerably smaller than the Lu2Cl6(g) contribution in LuCl3(s) vapors. The measurements were supplemented by quantum chemical ab initio studies of structures, energetics, and vibrational frequencies of neutral and singly ionized LnCl, LnCl2, and LnCl3 species (Ln = Ce, Lu). The theoretical appearance energies of different ions, calculated from the energies of the gaseous species, are in good agreement with experimental data. The fragmentation energies of LnCl, LnCl2, and LnCl3 were also computed and compared with the mass spectral patterns of respective vapor species. The Mulliken and natural bond orbital electron population methods were applied for the systematic analysis of the bonding scheme in molecules and cations. PMID- 16711700 TI - Di-, tri-, and tetranuclear zinc hydroxamate complexes as structural models for the inhibition of zinc hydrolases by hydroxamic acids. AB - Attempts to produce Zn analogues of the structural model complexes [M2(mu O2CR)2(O2CR)2(mu-H2O)(tmen)2] (M = Ni, Co, Mn; R = CH(3), C(CH3)3, CF3) by the reaction of a series of zinc carboxylates with N,N,N',N' tetramethylethylenediamine (tmen), resulted in the mononuclear complexes [Zn(OAc)(2)(tmen)] (1) and [Zn(crot)2(tmen)].(0.5)H2O (2) for R = CH3 and (CH)2CH3, respectively, and the dinuclear complexes [Zn(2)(mu-piv)(2)(piv)(2)(mu H2O)(tmen)2] (3) and [Zn2(mu-OAc(F))2(OAc(F))2(mu-H2O)(tmen)2] (4) for R = C(CH3)3 and CF3, respectively. In contrast to the analogous imidazole series, i.e., [M2(mu-O2CR)2(O2CR)2(mu-H2O)(Im)4] (M = Ni, Co, Mn; R = CH3, C(CH3)3, CF3), zinc carboxylates react with imidazole to give only the mononuclear complexes [Zn(OAc)2(Im)2] (5), [Zn(crot)2(Im)2].H2O (6), [Zn(piv)2(Im)2].(0.5)H2O (7), and [Zn(OAc(F))2(Im)2] (8). Reaction of 1, 2, and 3 with either acetohydroxamic acid (AHA) or benzohydroxamic acid (BHA) gives the dinuclear complexes [Zn2(O2CR)3(R'A)(tmen)], where R'A = acetohydroxamate (AA) (9, 10, 11) or benzohydroxamate (BA) (13, 14, 15). In these complexes, the zinc atoms are bridged by a single hydroxamate and two carboxylates, with a capping tmen ligand on one zinc and a monodentate carboxylate bonded to the second zinc atom. This composition models closely the observed structure of the active site of the p iodo-d-phenylalanine hydroxamic acid inhibited Aeromonas proteolyticaaminopeptidase enzyme. In contrast, 4 reacts with AHA to give [Zn2(OAc(F))3(tmen)2(AA)] (12) with an additional tmen ligand so that both Zn atoms are 6-coordinate, whereas reaction with BHA gives the trinuclear complex [Zn3(OAc(F))4(tmen)2(BA)2] (16). Reactions of 3 and 4 with glutarodihydroxamic acid (GluH2A2) produce the tetranuclear complexes [Zn4(piv)6(tmen)4(GluA2)] (18) and [Zn4(OAc(F))6(tmen)4(GluA2)] (19). PMID- 16711702 TI - X-ray crystallographic and Mossbauer spectroscopic applications in dependence of partial quadrupole splitting, [R], on the C-Sn-C angle in seven-coordinated diorganotin(IV) complexes. AB - The synthesis and the IR, NMR (1H, 13C, and 119Sn), and Mossbauer spectroscopies and single-crystal X-ray diffraction studies of seven-coordinated diorganotin(IV) complexes, namely, [Ph2Sn(Hdapsc)]Cl.H2O.DMF [7; H(2)dapsc = 2,6-diacetylpyridine bis(semicarbazone)], [Me(2)Sn(H2,6Achexim)]Br.H2O [8; H(2)2,6Achexim = 2,6 diacetylpyridine bis(3-hexamethyleneiminylthiosemicarbazone)], [Me(2)Sn(dapmts)] [9; H(2)dapmts = 2,6-diacetylpyridine bis(4-methythiosemicarbazone)], and [nBu2Sn(dapmdtc)] [10; H(2)dapmdtc = 2,6-diacetylpyridine bis(S methydithiocarbazate)], were done. The determination of the structures of [Ph(2)Sn(Hdapsc)]+, [Me2Sn(H2,6Achexim)]+ and [Me2Sn(dapmts)], [nBu2Sn(dapmdtc)] revealed the presence of monocationic and neutral complexes, respectively. The structures consist of monomeric units in which the Sn(IV) ions exhibit distorted pentagonal-bipyramidal geometries, with the X,N,N,N,X-donor (X = O, S) systems of the ligands lying in the equatorial plane and the organic groups in the apical positions. The C-Sn-C angle in the seven-coordinated diorganotin(IV) complexes was estimated using a correlation between Mossbauer and X-ray data based on the point-charge model and using new values obtained in this work for [alkyl] = -1.00 mm s(-1) and [aryl] = -0.80 mm s(-1) for complexes containing O,N,N,N,O pentadentate ligands and new values for [alkyl] = -0.87 mm s(-1) and [aryl] = 0.75 mm s(-1) for complexes containing S,N,N,N,S-pentadentate ligands. PMID- 16711703 TI - Spectral and structural characterization of amidate-bridged platinum-thallium complexes with strong metal-metal bonds. AB - The reactions of [Pt(NH3)2(NHCOtBu)2] and TlX3 (X = NO3-, Cl-, CF3CO2-) yielded dinuclear [{Pt(ONO2)(NH3)2(NHCOtBu)}Tl(ONO2)2(MeOH)] (2) and trinuclear complexes [{PtX(RNH2)2(NHCOtBu)2}2Tl]+ [X = NO3- (3), Cl- (5), CF3CO2- (6)], which were spectroscopically and structurally characterized. Strong Pt-Tl interaction in the complexes in solutions was indicated by both 195Pt and 205Tl NMR spectra, which exhibit very large one-bond spin-spin coupling constants between the heteronuclei (1J(PtTl)), 146.8 and 88.84 kHz for 2 and 3, respectively. Both the X-ray photoelectron spectra and the 195Pt chemical shifts reveal that the complexes have Pt centers whose oxidation states are close to that of Pt(III). Characterization of these complexes by X-ray diffraction analysis confirms that the Pt and Tl atoms are held together by very short Pt-Tl bonds and are supported by the bridging amidate ligands. The Pt-Tl bonds are shorter than 2.6 Angstrom, indicating a strong metal-metal attraction between these two metals. Compound 2 was found to activate the C-H bond of acetone to yield a platinum(IV) acetonate complex. This reactivity corresponds to the property of Pt(III) complexes. Density functional theory calculations were able to reproduce the large magnitude of the metal-metal spin-spin coupling constants. The couplings are sensitive to the computational model because of a delicate balance of metal 6s contributions in the frontier orbitals. The computational analysis reveals the role of the axial ligands in the magnitude of the coupling constants. PMID- 16711704 TI - Novel stoichiometrically erbium-ytterbium cocrystalline complex exhibiting enhanced near-infrared luminescence. AB - Three beta-diketonate complexes of erbium and ytterbium, Er(HFA)3(TPPO)3, Yb(HFA)3(TPPO)3, and Er(1/2)Yb(1/)(2)(HFA)3(TPPO)2 (HFA = hexafluoroacetylacetonate, TPPO = triphenylphosphine oxide) are synthesized and structurally characterized, and their luminescent properties are examined in detail. Compared with monometallic complexes Er(HFA)3(TPPO)3 and Er(HFA)3(TPPO)3, the stoichiometrically Er-Yb cocrystalline complex Er(1/2)Yb(1/2)(HFA)3(TPPO)2 exhibits special indirect near-infrared (NIR) luminescence with the acceptor and sensitizer (donor) coexisting in a single crystal. Of interest is that the NIR luminescence of the erbium ion in Er(1/2)Yb(1/2)(HFA)3(TPPO)2 is significantly enhanced because of the coexistence of the ytterbium ion through the energy transfer between the ytterbium and erbium ions. The estimated donor-acceptor energy-transfer critical radii R(C,DA), obtained from the Forster model, are in good agreement with those determined from the single X-ray crystal structure, confirming the dipole-dipole character for such a Yb-Er energy-transfer process. PMID- 16711706 TI - Molybdenum oxo-imido aryloxide complexes: oxo analogues of olefin metathesis catalysts. AB - The novel 16-electron molybdenum oxo-imido bis(aryloxide) complexes [Mo(NtBu)(O)(2,6-Me2C6H3O)2(py)] (1) and [Mo(NtBu)(O)(2,6-iPr2C6H3O)2(py)] (2) have been prepared by the salt elimination reactions of [Mo(NtBu)(O)Cl2(DME)] with the appropriate lithium aryloxide and from the cycloaddition reactions of tert-butyl isocyanate with the appropriate molybdenum dioxo bis(aryloxide) complex [Mo(O)2(OAr)2(py)n]. Complexes 1 and 2 are the first isolable and crystallographically characterized molybdenum oxo-imido aryloxide complexes. The geometry around the metal in complexes 1 and 2 is best described as a distorted trigonal bipyramid, with the imido and pyridine ligands occupying the axial positions and the oxo and aryloxide ligands in the equatorial plane. X-ray and IR data have confirmed that the imido ligand is the dominant pi donor in the complexes, resulting in an Mo-O bond order of less than 2.5. Reaction of [Mo(NtBu)(O)Cl2(DME)] with Li(OCH2tBu) instead gave the novel complex [Mo(NtBu)(OCH2tBu)3Cl(py)] (3). PMID- 16711705 TI - Probing the electronic structure of platinum(II) chromophores: crystal structures, NMR structures, and photophysical properties of six new bis- and di- phenolate/thiolate Pt(II)diimine chromophores. AB - A general route for synthesis of six structurally similar Pt(II) diimine thiolate/phenolates chromophores possessing bulky phenolate or thiolate ligands is reported. The Pt chromophores were characterized using an array of techniques including 1H, 13C, and 195Pt NMR, absorption, emission, (spectro)electrochemistry, and EPR spectroscopy. Systematic variation of the electronic structure of the Pt(II) chromophores studied was achieved by (i) changing solvent polarity; (ii) substituting oxygen for sulfur in the donor ligand; (iii) alternating donor ligands from bis- to di-coordination; and (iv) changing the electron donating/withdrawing properties of the ligand(s). The lowest excited state in these new chromophores was assigned to a [charge-transfer to-diimine] transition from the HOMO of mixed Pt/S (or Pt/O) character on the basis of absorption and emission spectroscopy, UV/vis (spectro)electrochemistry, and EPR spectroscopy. One of the chromophores, Pt(dpphen)(3,5-di-tert-butyl catecholate) represents an example of a Pt(II) diimine phenolate chromophore that possesses a reversible oxidation centered predominantly on the donor ligand. Results from EPR spectroscopy indicate participation of the Pt(II) orbitals in the HOMO. There is a dramatic difference in the photophysical properties of carborane complexes compared to other mixed-ligand Pt(II) compounds, which includes room-temperature emission and photostability. The charge-transfer character of the lowest excited state in this series of chromophores is maintained throughout. Moreover, the absorption and emission energies and the redox properties of the excited state can be significantly tuned. PMID- 16711707 TI - Rh(I) and Ir(i) derivatives of a P(S),N-substituted indene ligand: synthetic, structural, and catalytic alkene hydrosilylation studies. AB - Treatment of 1-PiPr2-indene or 1-PiPr2-2-NMe2-indene (1a) with elemental sulfur afforded 3-iPr2P(S)-indene or 1-iPr2P(S)-2-NMe2-indene (4a) in 81% and 85% isolated yield, respectively. Addition of 4a to [(COD)M(THF)2]+BF4- afforded the corresponding [(COD)M(kappa2-N,S-4a)]+BF4- complexes (M = Rh, 5a, 76%; M = Ir, 5b, 59%; COD = eta4-1,5-cyclooctadiene), which were found to exhibit temperature dependent NMR spectral features that were rationalized in terms of a dynamic process involving M-NMe2 dissociation, rotation about the indenyl-NMe2 bond, inversion at nitrogen, and re-coordination to M. Analysis of variable-temperature NMR data collected for 5a and 5b each yielded a value for DeltaG(double dagger) of ca. 14 kcal/mol for this process. Exposure of 5a or 5b to NaN(SiMe3)2 generated the corresponding (COD)M(kappa2-C,S-1-iPr2P(S)-2-NMe2-(C1-indenyl)) complex (M = Rh, 6a, 70%; M = Ir, 6b, 86%) in which the metal is incorporated into an M-C-P-S ring via coordination to the indenyl ring in an eta1-fashion, as well as to sulfur. Alternatively, complex 6b was prepared cleanly via lithiation of 4a followed by treatment with 0.5 equiv of [(COD)IrCl]2. The ability of 5a,b and 6a,b to mediate the addition of triethylsilane to styrene was also explored, and their performance was compared with that of Wilkinson's Catalyst ((PPh3)3RhCl) and Crabtree's catalyst ([(COD)Ir(PCy3)(Py)]+PF6-; Cy = cyclohexyl; Py = pyridine). Single-crystal X-ray diffraction data are provided for 4a, 2-NMe2 3-iPr2P(S)-indene (4b), 6a, and 6b. PMID- 16711712 TI - Bringing chemical data onto the Semantic Web. AB - Present chemical data storage methodologies place many restrictions on the use of the stored data. The absence of sufficient high-quality metadata prevents intelligent computer access to the data without human intervention. This creates barriers to the automation of data mining in activities such as quantitative structure-activity relationship modelling. The application of Semantic Web technologies to chemical data is shown to reduce these limitations. The use of unique identifiers and relationships (represented as uniform resource identifiers, URIs, and resource description framework, RDF) held in a triplestore provides for greater detail and flexibility in the sharing and storage of molecular structures and properties. PMID- 16711708 TI - Thiolate alkylation in tripod zinc complexes: a comparative kinetic study. AB - The biologically relevant alkylations of the thiolate ligands in tripod zinc thiolates by methyl iodide were studied kinetically. Five tripod ligands of the pyrazolyl/thioimidazolyl borate type were employed, offering N3, N2S, NS2, and S3 donor sets. For each of them, the ethyl-, benzyl-, phenyl-, and p nitrophenylthiolate zinc complexes were investigated, yielding a total of 20 second-order rate constants. The comparison of these rate constants shows three effects: (1) the electronic effect among the thiolates, i.e., the ethanethiolates react about 3 orders of magnitude faster than the p-nitrophenylthiolates; (2) the steric effect among the pyrazolylborates, i.e., the phenyl-substituted ones react about 2 orders of magnitude faster than the tert-butyl-substituted ones; and (3) the strong acceleration by the sulfur donors in the tripods, reaching 4 orders of magnitude between the reaction times of the (N3)Zn-SR and (S3)Zn-SR complexes. PMID- 16711713 TI - Open computing grid for molecular science and engineering. AB - Grid is an emerging infrastructure for distributed computing that provides secure and scalable mechanisms for discovering and accessing remote software and data resources. Applications built on this infrastructure have great potential for addressing and solving large scale chemical, pharmaceutical, and material science problems. The article describes the concept behind grid computing and will present the OpenMolGRID system that is an open computing grid for molecular science and engineering. This system provides grid enabled components, such as a data warehouse for chemical data, software for building QSPR/QSAR models, and molecular engineering tools for generating compounds with predefined chemical properties or biological activities. The article also provides an overview about the availability of chemical applications in the grid. PMID- 16711714 TI - A computer-aided drug discovery system for chemistry teaching. AB - The Schools Malaria Project (http://emalaria.soton.ac.uk/) brings together school students with university researchers in the hunt for a new antimalaria drug. The design challenge being offered to students is to use a distributed drug search and selection system to design potential antimalaria drugs. The system is accessed via a Web interface. This e-science project displays the results of the trials in an accessible manner, giving students an opportunity for discussion and debate both with peers and with the university contacts. The project has been implemented by using distributed computing techniques, spreading computer load over a network of machines that cross institutional boundaries, forming a grid. This provides access to greater computing power and allows a much more complex and detailed formulation of the drug design problem to be tackled for research, teaching, and learning. PMID- 16711715 TI - Computational Science and Engineering Online (CSE-Online): a cyber-infrastructure for scientific computing. AB - With the expansion of the Internet and World Wide Web (or the Web), research environments have changed dramatically. As a result, the need to be able to efficiently and securely access information and resources from remote computer systems is becoming even more critical. This paper describes the development of an extendable integrated Web-accessible simulation environment for computational science and engineering called Computational Science and Engineering Online (CSE Online; http://cse-online.net). CSE-Online is based on a unique client-server software architecture that can distribute the workload between the client and server computers in such a way as to minimize the communication between the client and server, thus making the environment less-sensitive to network instability. Furthermore, the new software architecture allows the user to access data and resources on one or more remote servers as well as on the computing grid while having the full capability of the Web-services collaborative environment. It can be accessed anytime and anywhere from a Web browser connected to the network by either a wired or wireless connection. It has different modes of operations to support different working environments and styles. CSE-Online is evolving into middleware that can provide a framework for accessing and managing remote data and resources including the computing grid for any domain, not necessarily just within computational science and engineering. PMID- 16711716 TI - ChemSem: an extensible and scalable RSS-based seminar alerting system for scientific collaboration. AB - A seminar announcement system based on the extensive use of XML-based data structures, CML/MathML for carrying more domain-specific molecular content, and open source software components is described. The output is a resource description framework (RDF) site summary (RSS) feed, which potentially carries many advantages over conventional announcement mechanisms, including the ability to aggregate and then sort multiple and diverse RSS feeds on the basis of declared metadata and to feed into RDF-based mechanisms for establishing links between different subject areas. PMID- 16711717 TI - The Blue Obelisk-interoperability in chemical informatics. AB - The Blue Obelisk Movement (http://www.blueobelisk.org/) is the name used by a diverse Internet group promoting reusable chemistry via open source software development, consistent and complimentary chemoinformatics research, open data, and open standards. We outline recent examples of cooperation in the Blue Obelisk group: a shared dictionary of algorithms and implementations in chemoinformatics algorithms drawing from our various software projects; a shared repository of chemoinformatics data including elemental properties, atomic radii, isotopes, atom typing rules, and so forth; and Web services for the platform-independent use of chemoinformatics programs. PMID- 16711718 TI - High-throughput calculation of protein-ligand binding affinities: modification and adaptation of the MM-PBSA protocol to enterprise grid computing. AB - We have developed a system for performing computations on an enterprise grid using a freely available package for grid computing that allows us to harvest unused CPU cycles off of employee desktop computers. By modifying the traditional formulation of Molecular Mechanics with Poisson-Boltzmann Surface Area (MM-PBSA) methodology, in combination with a coarse-grain parallelized implementation suitable for deployment onto our enterprise grid, we show that it is possible to produce rapid physics-based estimates of protein-ligand binding affinities that have good correlation to experimental data. This is demonstrated by examining the correlation of our calculated binding affinities to experimental data and also by comparison to the correlation obtained from the binding-affinity calculations using traditional MM-PBSA that are reported in the literature. PMID- 16711719 TI - An e-science environment for service crystallography--from submission to dissemination. AB - The U.K. National Crystallography Service (NCS) has developed a prototype e science infrastructure for the provision of a small molecule crystallography service from sample receipt to results dissemination. This paper outlines the two strands of this service, which (a) enable a user to contribute in the conduction of an experiment and (b) provide an effective route for the archival and dissemination of the arising results. Access to use the NCS facilities and expertise and a mechanism to submit samples is granted through a secure Grid infrastructure, which seamlessly provides instantaneous feedback and the ability to remotely monitor and guide diffraction experiments and stage the diffraction data to a securely accessible location. Publication of all the data and results generated during the course of the experiment, from processed data to analyzed structures, is then enabled by means of an open access data repository. The repository publishes its content through established digital libraries' protocols, which enable harvester and aggregator services to make the data searchable and accessible. PMID- 16711720 TI - Instrument monitoring, data sharing, and archiving using Common Instrument Middleware Architecture (CIMA). AB - The Common Instrument Middleware Architecture (CIMA) aims at Grid-enabling a wide range of scientific instruments and sensors to enable easy access to and sharing and storage of data produced by these instruments and sensors. This paper describes the implementation of CIMA applied to the field of single-crystal X-ray crystallography. To allow the researchers to easily view the current and past data streams from the instruments or sensors in a laboratory, a crystallography portal and associated portlets were developed for this application. The CIMA based crystallography system provides an opportunity for anyone with Web access to observe and use crystallographic and other data from laboratories that previously had only limited access. PMID- 16711721 TI - An integrated instrument control and informatics system for combinatorial materials research. AB - The use of high-throughput synthesis and characterization techniques is increasingly prevalent in materials science research. We describe the London University Search Instrument, a research apparatus designed for the high throughput synthesis and characterization of thick-film sample libraries of ceramic compounds. The instrument is constructed largely from commodity components, which pose particular engineering challenges for achieving the automated operation required for efficient high-throughput experimentation. This paper describes the architecture and implementation of the software system that provides integrated instrument control and data management functions. PMID- 16711722 TI - A database-centric virtual chemistry system. AB - We describe an Oracle database application for general use within virtual chemistry. The application functions as a central hub and repository for chemical data with interfaces to external calculators. It deals with the general problems of merging data from disparate sources and with scheduling of computational tasks for parallel or sequential execution in a mixed environment. The central database is used for the storage of input, intermediary, and final data as well as for job control. A calculation job is split into distinct tasks, or units of work, which are put in a queue. Tasks are dequeued and handled by specialized calculators. These calculators are in-house or commercial programs for which adaptor modules for connection to the database must be written. Tasks are handled in a transactional fashion, so that uncompleted or failed tasks are left in the queue. This makes the system stable to many types of disturbances. Sorting, filtering, and merging operations are handled by the database itself. Usage is very general, but some specific examples are (1) as a back end for a chemical property calculator Web page, (2) in an automated quantitative structure-activity relationship system, and (3) in virtual screens. PMID- 16711723 TI - COSMOsim: bioisosteric similarity based on COSMO-RS sigma profiles. AB - A novel approach for the quantification of drug similarity is proposed, which makes use of the surface polarities, that is, conductor surface polarization charge densities sigma, as defined in the quantum chemically based conductor-like screening model for realistic solvation(COSMO-RS). The histogram of these surface polarities, the so-called sigma profiles, have been proven to be the key for the calculation of all kinds of partition and adsorption coefficients and, therefore, of relevant absorption, distribution, metabolism, and excretion parameters such as solubility, pKa, log BB, and many others. They also carry a large part of the information required for the estimation of desolvation and binding processes responsible for receptor binding and enzyme inhibition of drug molecules. Thus, a large degree of similarity with respect to the sigma profiles appears to be a necessary condition for drugs of similar physiological action. Driven by this insight, we propose a sigma-profile-based drug similarity measure COSMOsim for the detection of new bioisosteric drug candidates. In several examples, we demonstrate its statistical and pharmaceutical plausibility, its practicability for real drug research projects, and its unique independence from the chemical structure, which enables scaffold hopping in a natural way. PMID- 16711724 TI - Visualization of large-scale aqueous solubility data using a novel hierarchical data visualization technique. AB - It is a difficult task to recognize the trends in molecular physical properties relevant to a specific chemical class and find a way to optimize potential compounds. We present here a novel hierarchical data visualization technique, named "HeiankyoView", to visualize large-scale multidimensional chemical information. HeiankyoView represents hierarchically organized data objects by mapping leaf nodes as colored square icons and nonleaf nodes as rectangular borders. In this way, data objects can be expressed as equishaped icons without overlapping one another in the two-dimensional display space. HeiankyoView has been applied to visualize aqueous solubility data for 908 compounds collected from the published literature. When the results of a recursive partitioning analysis and hierarchical clustering analysis were visualized, the trends hidden in the solubility data could be effectively displayed as intuitively understandable visual images. Most interestingly, the data visualization technique, without any statistical computations, was able to assist us in extracting from such large-scale data meaningful information establishing that ClogP and the molecular weight are critical factors in determining aqueous solubility. Thus, HeiankyoView is a powerful tool to help us understand structure activity relationships intuitively from a large-scale data set. PMID- 16711725 TI - An empirical process for the design of high-throughput screening deck filters. AB - A process for objective identification and filtering of undesirable compounds that contribute to high-throughput screening (HTS) deck promiscuity is described. Two methods of mapping hit promiscuity have been developed linking SMARTS-based structural queries with historical primary HTS data. The first compares an expected assay hit rate to actual hit rates. The second examines the propensity of an individual compound to hit multiple assays. Statistical evaluation of the data indicates a correlation between the resultant functional group filters and compound promiscuity. These data corroborate a number of commonly applied filters as well as producing some unexpected results. Application of these models to HTS collection triage reduced the number of in-house compounds considered for screening by 12%. The implications of these findings are further discussed in the context of the HTS screening set and combinatorial library design as well as compound acquisition. PMID- 16711726 TI - A family of ring system-based structural fragments for use in structure-activity studies: database mining and recursive partitioning. AB - In earlier work from our laboratory, we have described the use of the ring system and ring scaffold as descriptors. We showed that these descriptors could be used for fast compound clustering, novelty determination, compound acquisition, and combinatorial library design. Here we extend the concept to a whole family of structural descriptors with the ring system as the centerpiece. We show how this simple idea can be used to build powerful search tools for mining chemical databases in useful ways. We have also built recursive partition trees using these fragments as descriptors. We will discuss how these trees can help in analyzing complex structure-activity data. PMID- 16711727 TI - Analysis of activity space by fragment fingerprints, 2D descriptors, and multitarget dependent transformation of 2D descriptors. AB - The effect of multitarget dependent descriptor transformation on classification performance is explored in this work. To this end decision trees as well as neural net QSAR in combination with PLS were applied to predict the activity class of 5HT3 ligands, angiotensin converting enzyme inhibitors, 3-hydroxyl-3 methyl glutaryl coenzyme A reductase inhibitors, platelet activating factor antagonists, and thromboxane A2 antagonists. Physicochemical descriptors calculated by MOE and fragment-based descriptors (MOLPRINT 2D) were employed to generate descriptor vectors. In a subsequent step the physicochemical descriptor vectors were transformed to a lower dimensional space using multitarget dependent descriptor transformation. Cross-validation of the original physicochemical descriptors in combination with decision trees and neural net QSAR as well as cross-validation of PLS multitarget transformed descriptors with neural net QSAR were performed. For comparison this was repeated using fragment-based descriptors in combination with decision trees. PMID- 16711728 TI - Root mean square deviation probability analysis of molecular dynamics trajectories on DNA. AB - The comparison and detection of the commonalities and differences in multiple structural ensembles is an important step in the use of molecular simulations to gain insight into the conformation and dynamics of complex biomacromolecules. While the average structure is often employed as the representative of an ensemble of structures in such comparisons, dynamic molecular systems with multiple conformational substates call for a more accurate representation that captures the complete dynamical range of the ensemble. We present a probability analysis procedure based on the root-mean-square differences among the structural ensembles that efficiently and accurately performs the relevant comparison. PMID- 16711729 TI - A distance function for retrieval of active molecules from complex chemical space representations. AB - The concept of chemical space is of fundamental importance for chemoinformatics research. It is generally thought that high-dimensional space representations are too complex for the successful application of many compound classification or virtual screening methods. Here, we show that a simple "activity-centered" distance function is capable of accurately detecting molecular similarity relationships in "raw" chemical spaces of high dimensionality. PMID- 16711730 TI - Assessing different classification methods for virtual screening. AB - How well do different classification methods perform in selecting the ligands of a protein target out of large compound collections not used to train the model? Support vector machines, random forest, artificial neural networks, k-nearest neighbor classification with genetic-algorithm-optimized feature selection, trend vectors, naive Bayesian classification, and decision tree were used to divide databases into molecules predicted to be active and those predicted to be inactive. Training and predicted activities were treated as binary. The database was generated for the ligands of five different biological targets which have been the object of intense drug discovery efforts: HIV-reverse transcriptase, COX2, dihydrofolate reductase, estrogen receptor, and thrombin. We report significant differences in the performance of the methods independent of the biological target and compound class. Different methods can have different applications; some provide particularly high enrichment, others are strong in retrieving the maximum number of actives. We also show that these methods do surprisingly well in predicting recently published ligands of a target on the basis of initial leads and that a combination of the results of different methods in certain cases can improve results compared to the most consistent method. PMID- 16711731 TI - 2D structure depiction. AB - A comprehensive algorithm for the depiction of 2D coordinates of chemical structures is described. The methods used represent a significant improvement to the state of the art with regard to molecular connection graphs which pose particular difficulty to most layout efforts. Resulting coordinates are consistently of publication quality for a large subset of chemistry. The algorithm is discussed in detail, and measurements of its overall success are presented. PMID- 16711732 TI - Prediction of biological targets for compounds using multiple-category Bayesian models trained on chemogenomics databases. AB - Target identification is a critical step following the discovery of small molecules that elicit a biological phenotype. The present work seeks to provide an in silico correlate of experimental target fishing technologies in order to rapidly fish out potential targets for compounds on the basis of chemical structure alone. A multiple-category Laplacian-modified naive Bayesian model was trained on extended-connectivity fingerprints of compounds from 964 target classes in the WOMBAT (World Of Molecular BioAcTivity) chemogenomics database. The model was employed to predict the top three most likely protein targets for all MDDR (MDL Drug Database Report) database compounds. On average, the correct target was found 77% of the time for compounds from 10 MDDR activity classes with known targets. For MDDR compounds annotated with only therapeutic or generic activities such as "antineoplastic", "kinase inhibitor", or "anti-inflammatory", the model was able to systematically deconvolute the generic activities to specific targets associated with the therapeutic effect. Examples of successful deconvolution are given, demonstrating the usefulness of the tool for improving knowledge in chemogenomics databases and for predicting new targets for orphan compounds. PMID- 16711733 TI - The influence of variations of ligand protonation and tautomerism on protein ligand recognition and binding energy landscape. AB - We investigate the influence of variations of ligand protonation and tautomeric states on the protein-ligand binding energy landscape by applying the concept of structural consensus. In docking simulations, allowing full flexibility of the ligand, we explore whether the native binding mode could be successfully recovered using a non-native ligand protonation state. Here, we consider three proteins, dihydrofolate reductase, transketolase, and alpha-trichosanthin, complexed with ligands having multiple tautomeric forms. We find that for the majority of protonation and tautomeric states the native binding mode can be recovered without a great loss of accuracy. PMID- 16711734 TI - Hard modeling methods for the curve resolution of data from liquid chromatography with a diode array detector and on-flow liquid chromatography with nuclear magnetic resonance spectroscopy. AB - Hard modeling methods have been performed on data from high-performance liquid chromatography with a diode array detector (LC-DAD) and on-flow liquid chromatography with 1H nuclear magnetic spectroscopy (LC-NMR). Four methods have been used to optimize parameters to model concentration profiles, three of which belong to classical optimization methods (the simplex method of Nelder-Mead, sequential quadratic programming approach, and Levenberg-Marquardt method), and the fourth is the application of genetic algorithms using real-value encoding. Only classical methods worked well for LC-DAD data, while all of the methods produced good results when LC-NMR data were divided into small spectral windows of peak clusters and parameters were optimized over each window. PMID- 16711735 TI - Hierarchical PLS modeling for predicting the binding of a comprehensive set of structurally diverse protein-ligand complexes. AB - A new approach is presented for predicting ligand binding to proteins using hierarchical partial-least-squares regression to latent structures (Hi-PLS). Models were based on information from the 2002 release of the PDBbind database containing (after in-house refinement) high-resolution X-ray crystallography and binding affinity (Kd or Ki) data for 612 protein-ligand complexes. The complexes were characterized by four different descriptor blocks: three-dimensional (3D) structural descriptors of the proteins, protein-ligand interactions according to the Validate scoring function, binding site surface areas, and ligand 2D and 3D descriptors. These descriptor blocks were used in Hi-PLS models, generated using both linear and nonlinear terms, to relate the characterizations to pKd/i. The results show that each of the four descriptor blocks contributed to the model, and the predictions of pKd/i of the internal test set gave a root-mean-square error of prediction (RMSEP) of 1.65. The data were further divided according to the structural classification of the proteins, and Hi-PLS models were constructed for the resulting subclasses. The models for the four subclasses differed considerably in terms of both their ability to predict pKd/i (with RMSEPs ranging from 0.8 to 1.56) and the descriptor block that had the strongest influence. The models were validated with an external test set of 174 complexes from the 2003 release of the PDBbind database. The overall results show that the presented Hi PLS methodology could facilitate the difficult task of predicting binding affinity. PMID- 16711736 TI - Exhaustive de novo design of low-molecular-weight fragments against the ATP binding site of DNA-gyrase. AB - We present a de novo design approach to generating small fragments in the DNA gyrase ATP-binding site using the computational drug design platform SkelGen. We have generated an exhaustive number of structural possibilities, which were subsequently filtered for site complementarity and synthetic tractability. A number of known active fragments are found, but most of the species created are potentially novel and could be valuable for further elaboration and development into lead-like structures. PMID- 16711737 TI - A QXP-based multistep docking procedure for accurate prediction of protein-ligand complexes. AB - The two great challenges of the docking process are the prediction of ligand poses in a protein binding site and the scoring of the docked poses. Ligands that are composed of extended chains in their molecular structure display the most difficulties, predominantly because of the torsional flexibility. On the basis of the molecular docking program QXP-Flo+0802, we have developed a procedure particularly for ligands with a high degree of rotational freedom that allows the accurate prediction of the orientation and conformation of ligands in protein binding sites. Starting from an initial full Monte Carlo docking experiment, this was achieved by performing a series of successive multistep docking runs using a local Monte Carlo search with a restricted rotational angle, by which the conformational search space is limited. The method was established by using a highly flexible acetylcholinesterase inhibitor and has been applied to a number of challenging protein-ligand complexes known from the literature. PMID- 16711738 TI - Structural unit analysis identifies lead series and facilitates scaffold hopping in combinatorial chemistry. AB - Combinatorial chemistry and high-throughput screening technologies produce huge amounts of data on a regular basis. Sieving through these libraries of compounds and their associated assay data to identify appropriate series for follow-up is a daunting task, which has created a need for computational techniques that can find coherent islands of structure-activity relationships in this sea. Structural unit analysis (SUA) examines an entire data set so as to identify the molecular substructures or fragments that distinguish compounds with high activity from those with average activity. The algorithm is iterative and follows set heuristics in order to generate the structural units. It produces graphs that represent a set of units, which become SUA rules. Finding all of the input structures that match these graphs generates clusters. The Apriori algorithm for association rule mining is adapted to explore all of the combinations of structural units that define useful series. User-defined constraints are applied toward series selection and the refinement of rules. The significance of a series is determined by applying statistical methods appropriate to each data set. Application to the NCI-H23 (DTP Human Tumor Cell Line Screen) database serves to illustrate the process by which structural series are identified. An application of the method to scaffold hopping is then discussed in connection with proprietary screening data from a lead optimization project directed toward the treatment of respiratory tract infections at Bayer Healthcare. SUA was able to successfully identify promising alternative core structures in addition to identifying compounds with above-average activity and selectivity. PMID- 16711740 TI - Multifingerprint based similarity searches for targeted class compound selection. AB - Molecular fingerprints are widely used for similarity-based virtual screening in drug discovery projects. In this paper we discuss the performance and the complementarity of nine two-dimensional fingerprints (Daylight, Unity, AlFi, Hologram, CATS, TRUST, Molprint 2D, ChemGPS, and ALOGP) in retrieving active molecules by similarity searching against a set of query compounds. For this purpose, we used biological data from HTS screening campaigns of four protein families (GPCRs, kinases, ion channels, and proteases). We have established threshold values for the similarity index (Tanimoto index) to be used as starting points for similarity searches. Based on the complementarities between the selections made by using different fingerprints we propose a multifingerprint approach as an efficient tool to balance the strengths and weaknesses of various fingerprints. PMID- 16711739 TI - Gas-phase and solution conformations of the alpha-L-iduronic acid structural unit of heparin. AB - The IdoA2S structural unit of heparin (subunit G) may oscillate among the three conformations (4C1, 1C4, and 2So). Only the twisted boat conformation allowed the biologically active pentasaccharide unit of heparin (DEFGH) to bind to antithrombin. Our work reports, in detail, the results of systematic large-scale theoretical investigations of the three basic conformations (4C1, 1C4, and 2So) of the IdoA2S structural unit of heparin, its anionic forms, and its sodium salt using the B3LYP/6-311++G(d, p) and B3LYP/6-31+G(d) model chemistries. According to our calculations, the most stable structure of these molecules corresponds to the 2So skew-boat conformation. This form is also the most stable in a water solution. The 2So conformation of neutral molecules is not maintained in the anionic species. With anions, both 1C4 and 4C1 conformations are present. The relative stability of individual species of the substituted iduronic acid affects extra stabilization by means of intramolecular hydrogen bonds. The calculated macroscopic pKa of 1,4-DiOMe IdoA2S are as follows: pKa = 0.25 for the terminal C(2)-OSO3H group, pKa = 3.67 for the terminal C(5)-CO2H group, and pKa = 14.00 for the C(3)-OH hydroxyl group. The computed Gibbs interaction energies, DeltaGdegrees , for the reaction 1,4-DiOMe IdoA2S(2-) + 2Na+ <==> 1,4-DiOMe IdoA2SNa2 (4C1, 1C4, and 2So conformations) are negative and span a rather small energy interval (from -1244 to -1290 kJ mol(-1)). PMID- 16711741 TI - Can 'Bacterial-Metabolite-Likeness' model improve odds of 'in silico' antibiotic discovery? AB - 'Inductive' QSAR descriptors have been used to develop the series of QSAR models enabling 'in silico' distinguishing between antimicrobial compounds, conventional drugs, and druglike substances. The constructed neural network-based models operating by 30 'inductive' parameters have been validated on an extensive set of 2686 chemical structures and resulted in up to 97% accurate separation of the three types of molecular activities. The demonstrated ability of 'inductive' parameters to adequately capture molecular features determining 'antibiotic-like' and 'druglike' potentials have been further utilized to construct a model of 'Bacterial-Metabolite-Likeness' (BML). The same 'inductive' descriptors have been used to train a neural network that could very accurately recognize substances involved into bacterial metabolism (that have been experimentally identified). When the developed model has been applied to the mixed set of antimicrobials, drugs, and druglike chemicals (not used for training the BML model), it exhibited a 2-5-fold recognition preference toward antimicrobial compounds compared to general drugs and an 18- to 45-fold preference when compared to a druglike substance (depending on the model stringency). These results illustrate immanent similarity between conventional antimicrobials and native bacterial metabolites and suggest that the developed BML model can be an effective classification tool for 'in silico' antibiotic studies. PMID- 16711742 TI - Modeling the structural basis of human CCR5 chemokine receptor function: from homology model building and molecular dynamics validation to agonist and antagonist docking. AB - This article describes the construction and validation of a three-dimensional model of the human CCR5 receptor using a homology-based approach starting from the X-ray structure of the bovine rhodopsin receptor. The reliability of the model is assessed through molecular dynamics and docking simulations using both natural agonists and a synthetic antagonist. Some important structural and functional features of the receptor cavity and the extracellular loops are identified, in agreement with data available from site-directed mutagenesis. The results of this study help to explain the structural basis for the recognition, activation, and inhibition processes of CCR5 and may provide fresh insights for the design of HIV-1 entry blockers. PMID- 16711743 TI - Enhancing specificity and sensitivity of pharmacophore-based virtual screening by incorporating chemical and shape features--a case study of HIV protease inhibitors. AB - Virtual screening (VS), if applied appropriately, could significantly shorten the hit identification and hit-to-lead processes in drug discovery. Recently, the version of VS that is based upon similarity to a pharmacophore has received increased attention. This is due to two major factors: first, the public availability of the ZINC1 conformational database has provided a large selection pool with high-quality and purchasable small molecules; second, new technology has enabled a more accurate and flexible definition of pharmacophore models coupled with an efficient search speed. The major goal of this study was to achieve improved specificity and sensitivity of pharmacophore-based VS by optimizing the variables used to generate conformations of small molecules and those used to construct pharmacophore models from known inhibitors or from inhibitor-protein complex structures. By using human immunodeficiency virus protease and its inhibitors (PIs) as a case study, the impact of the key variables, including the selection of chemical features, involvement of excluded volumes (EV), the tolerance radius of excluded volumes, energy windows, and the maximum number of conformers in conformation generation, was explored. Protein flexibility was simulated by adjusting the sizes of EV. Our best pharmacophore model, combining both chemical features and excluded volumes, was able to correctly identify 60 out of 75 structurally diverse known PIs, while misclassifying only 5 out of 75 similar compounds that are not inhibitors. To evaluate the specificity of the model, 1193 oral drugs on the market were screened, and 25 original hits were identified, including 5 out of 6 known PI drugs. PMID- 16711744 TI - Combinatorial QSAR modeling of P-glycoprotein substrates. AB - Quantitative structure-activity (property) relationship (QSAR/QSPR) models are typically generated with a single modeling technique using one type of molecular descriptors. Recently, we have begun to explore a combinatorial QSAR approach which employs various combinations of optimization methods and descriptor types and includes rigorous and consistent model validation (Kovatcheva, A.; Golbraikh, A.; Oloff, S.; Xiao, Y.; Zheng, W.; Wolschann, P.; Buchbauer, G.; Tropsha, A. Combinatorial QSAR of Ambergris Fragrance Compounds. J. Chem. Inf. Comput. Sci. 2004, 44, 582-95). Herein, we have applied this approach to a data set of 195 diverse substrates and nonsubstrates of P-glycoprotein (P-gp) that plays a crucial role in drug resistance. Modeling methods included k-nearest neighbors classification, decision tree, binary QSAR, and support vector machines (SVM). Descriptor sets included molecular connectivity indices, atom pair (AP) descriptors, VolSurf descriptors, and molecular operation environment descriptors. Each descriptor type was used with every QSAR modeling technique; so, in total, 16 combinations of techniques and descriptor types have been considered. Although all combinations resulted in models with a high correct classification rate for the training set (CCR(train)), not all of them had high classification accuracy for the test set (CCR(test)). Thus, predictive models have been generated only for some combinations of the methods and descriptor types, and the best models were obtained using SVM classification with either AP or VolSurf descriptors; they were characterized by CCR(train) = 0.94 and 0.88 and CCR(test) = 0.81 and 0.81, respectively. The combinatorial QSAR approach identified models with higher predictive accuracy than those reported previously for the same data set. We suggest that, in the absence of any universally applicable "one-for-all" QSAR methodology, the combinatorial QSAR approach should become the standard practice in QSPR/QSAR modeling. PMID- 16711745 TI - Amino Acid Sequence Autocorrelation vectors and ensembles of Bayesian-Regularized Genetic Neural Networks for prediction of conformational stability of human lysozyme mutants. AB - Development of novel computational approaches for modeling protein properties from their primary structure is a main goal in applied proteomics. In this work, we reported the extension of the autocorrelation vector formalism to amino acid sequences for encoding protein structural information with modeling purposes. Amino Acid Sequence Autocorrelation (AASA) vectors were calculated by measuring the autocorrelations at sequence lags ranging from 1 to 15 on the protein primary structure of 48 amino acid/residue properties selected from the AAindex database. A total of 720 AASA descriptors were tested for building predictive models of the thermal unfolding Gibbs free energy change of human lysozyme mutants. In this sense, ensembles of Bayesian-Regularized Genetic Neural Networks (BRGNNs) were used for obtaining an optimum nonlinear model for the conformational stability. The ensemble predictor described about 88% and 68% variance of the data in training and test sets, respectively. Furthermore, the optimum AASA vector subset was shown not only to successfully model unfolding thermal stability but also to distribute wild-type and mutant lysozymes on a stability Self-organized Map (SOM) when used for unsupervised training of competitive neurons. PMID- 16711746 TI - A quantum and deductive chemical study for all congeners of polybromo/chlorodibenzo-p-dioxin and polybromo/chlorodibenzofuran. AB - We have performed semiempirical quantum chemical calculations to obtain the optimized structure and the free energy (DeltaG) for all congeners (1701) of polybromo/chlorodibenzo-p-dioxins, which include all the isomers of all the homologues, and those for (3321) polybromo/chlorodibenzofurans. Then, to apply the "Quantum and Deductive Chemistry Approach" on the dioxin chemistry, we have carried out the multiple linear regression (MLR) as functions of temperature and the substituted numbers and positions of chlorine and bromine. The optimized structure of dibenzo-p-dioxin and the dibenzofuran ring is significantly influenced by the substitutions of the peri and lateral halogen atoms. The bond length between the aromatic ring and halogen atom also is influenced by the neighboring atoms. The bromine substitution at the 2 and 8 positions of dibenzofuran reduces the steric repulsion between the chlorine atoms at the 1 and 9 positions. The coefficients of the predicting equation of DeltaG derived by MLR suggest that the probabilities of chlorine elimination from the peri and lateral positions for polychlorodibenzo-p-dioxin are nearly equal. PMID- 16711747 TI - A quantum chemical study of the catalysis for cytidine deaminase: contribution of the extra water molecule. AB - Cytidine deaminase is known as an important enzyme responsible for the hydrolytic deamination of cytidine, which is applied as a key step to the conversion of the precursor of the cancer drug to an active form in the living body. Cytidine with water is efficiently converted to uridine with ammonia in the cleft of cytidine deaminase. In this work, the catalysis of cytidine deaminase for the hydrolytic deamination was examined using cytosine as a model of cytidine and the model molecules for the active site of cytidine deaminase by means of the quantum chemical method. We especially investigated the contribution of the water molecule from the solvent to the catalysis, because the X-ray diffraction analysis of a crystal structure has revealed the existence of the water molecule in the vicinity of the substrate bound to the active site inside the cleft. Our computations showed that the extra water molecule from the solvent has a possibility to support the catalysis of cytidine deaminase. PMID- 16711748 TI - WIN55212-2 docking to the CB1 cannabinoid receptor and multiple pathways for conformational induction. AB - Key pharmacophoric elements for the (aminoalkyl)indole (AAI) CB1 cannabinoid receptor agonists are the aminoalkyl moiety, the lipophilic aroyl group, and the heterocyclic indole ring. In the present study, the docking space allowed for (R) [2,3-dihydro-5-methyl-3-[(4-morpholinyl)methyl]pyrrolo[1,2,3-de]-1,4-benzoxazin-6 yl](1-naphthalenyl)methanone (WIN55212-2; 1) within the CB1 receptor was extensively explored by a docking approach that combines Monte Carlo (MC) and molecular dynamics (MD) simulations. The goals were to understand the key binding interactions of AAIs within the CB1 receptor and to examine the role of the ligand in inducing a receptor conformational change. From the findings of extensive SAR studies on the cannabinoid compounds and correlation between AAI binding affinity data and calculated binding energies, we proposed two alternative binding conformations, aroyl-up1 and aroyl-up2. These denote the directionality of the ligand naphthyl ring within the receptor upward with respect to the extracellular side. A comprehensive structural analysis of 1 demonstrated that the aroyl ring moiety could be important as the steric trigger for inducing CB1 receptor conformational change. Thus, it appears that aromatic aromatic interactions are important not only for the binding of 1 but also for inducing receptor conformational change. It is possible that differences in the nature of the ligand binding could contribute to ligand-specific conformational changes in the receptor. PMID- 16711749 TI - Pharmacophore modeling and in silico screening for new P450 19 (aromatase) inhibitors. AB - Cytochrome P450 19 (P450 19, aromatase) constitutes a successful target for the treatment of breast cancer. This study analyzes chemical features common to P450 19 inhibitors to develop ligand-based, selective pharmacophore models for this enzyme. The HipHop and HypoRefine algorithms implemented in the Catalyst software package were employed to create both common feature and quantitative models. The common feature model for P450 19 includes two ring aromatic features in its core and two hydrogen bond acceptors at the ends. The models were used as database search queries to identify active compounds from the NCI database. PMID- 16711750 TI - In silico renal clearance model using classical Volsurf approach. AB - A data set of 130 diverse compounds containing both central nervous system (CNS) and non-CNS drugs was used to generate a renal clearance model using a classical Volsurf approach. Percentage renal clearance data was used as a biological input. The score plots obtained from principal component analysis and partial least squares (PLS) analysis clearly separated high-clearance compounds from low clearance compounds. PLS models were used to predict the renal clearance of the data set. Categorical statistical methods such as SIMCA and recursive partitioning techniques were used for classifying the compounds into low- and high-clearance categories. PLS coefficient plots, Volsurf descriptor profiles, 3D Grid maps, and RP decision trees were used to explain the important descriptors separating low and high renal clearance compounds. For comparative purposes, topological descriptors such as Molconn-Z were also examined. All the models were validated by an external test set of 20 compounds. These models can be used as efficient tools in the classification and prediction of the renal clearance of unknown compounds, the knowledge of which is helpful in understanding their bioavailability behavior. PMID- 16711751 TI - Exploration of the conformational space of a polymeric material that inhibits human immunodeficiency virus. AB - Baertschi et al. (Antiviral Chem. Chemother. 1997, 8, 353-362) clarified the nature of a polymeric degradation product formed from the cephalosporin ceftazidime. Interest in the polymeric material arises from its ability to inhibit the RNase H and polymerase activities of HIV-1 reverse transcriptase (RT). To shed light on the structure of the polymeric material like that which forms from degradation of third-generation cephalosporins, we apply molecular modeling and other computational chemistry techniques. Aminothiazole methoxime (2 amino-4-thiazolyl-methoxyimino; ATMO) is the parent structure related to the isolated degradation product of ceftazidime. The MMFF94 force field and Monte Carlo multiple minimum method as implemented in MacroModel are used to generate low-energy conformers. We built up oligomeric models starting from the trimer to the 16-mer and performed distribution analyses on the dihedral angles from the Monte Carlo runs to analyze the three-dimensional shapes of the oligomers. Although the larger oligomers are too long for a complete search of conformational space, the low-energy conformers examined do not show secondary structure or repetitive conformations. Polymeric ATMO material may, therefore, exhibit only random coil conformations. Topological similarity of ATMO structures to other reported RT inhibitors is also examined. PMID- 16711753 TI - A combination of molecular dynamics and docking calculations to explore the binding mode of ADS-J1, a polyanionic compound endowed with anti-HIV-1 activity. AB - The HIV-1 entry process is an important target for the design of new pharmaceuticals for the multidrug therapy of AIDS. A lot of polyanionic compounds, such as polysulfonated and polysulfated, are reported in the literature for their ability to block early stages of HIV-1 replication. Several studies have been performed to elucidate the mechanism of the anti-HIV-1 activity of sulfated polysaccharides and polyanions in general, including binding to cell surface CD4 and interfering with the gp120-coreceptor interaction. Here, we show molecular modeling investigations on ADS-J1, a polyanionic compound with anti-HIV activity that is able to interfere with gp120-coreceptor interactions. Agreeing with experimental data, computer simulations suggested that the V3 loop of gp120 was the preferential binding site for ADS-J1 onto HIV-1. Moreover, mutations induced by the inhibitor significantly changed the stereoelectronic properties of the gp120 surface, justifying a marked drop in the affinity of ADS-J1 toward an ADS-J1-resistant HIV-1 strain. PMID- 16711752 TI - Ligand bias of scoring functions in structure-based virtual screening. AB - A total of 945 known actives and roughly 10 000 decoy compounds were docked to eight different targets, and the resulting poses were scored using 10 different scoring functions. Three different score postprocessing methods were evaluated with respect to improvement of the enrichment in virtual screening. The three procedures were (i) multiple active site correction (MASC) as has been proposed by Vigers and Rizzi, (ii) a variation of MASC where corrections terms are predicted from simple molecular descriptors through PLS, PLS MASC, and (iii) size normalization. It was found that MASC did not generally improve the enrichment factors when compared to uncorrected scoring functions. For some combinations of scoring functions and targets, the enrichment was improved, for others not. However, by excluding the standard deviation from the MASC equation and transforming the scores for each target to a mean of 0 and a standard deviation of 1 (unit variance normalization), the performance was improved as compared to the original MASC method for most combinations of targets and scoring functions. Furthermore, when the molecular descriptors were fit to the mean scores over all targets and the resulting PLS models were used to predict mean scores, the enrichment as compared to the raw score was improved more often than by straightforward MASC. A high to intermediate linear correlation between the score and the number of heavy atoms was found for all scoring functions except FlexX. There seems to be a correlation between the size dependence of a scoring function and the effectiveness of PLS MASC in increasing the enrichment for that scoring function. Finally, normalization by molecular weight or heavy atom count was sometimes successful in increasing the enrichment. Dividing by the square or cubic root of the molecular weight or heavy atom count instead was often more successful. These results taken together suggest that ligand bias in scoring functions is a source of false positives in structure-based virtual screening. The number of false positives caused by ligand bias may be decreased using, for example, the PLS MASC procedure proposed in this study. PMID- 16711754 TI - PharmID: pharmacophore identification using Gibbs sampling. AB - The binding of a small molecule to a protein is inherently a 3D matching problem. As crystal structures are not available for most drug targets, there is a need to be able to infer from bioassay data the key binding features of small molecules and their disposition in space, the pharmacophore. Fingerprints of 3D features and a modification of Gibbs sampling to align a set of known flexible ligands, where all compounds are active, are used to discern possible pharmacophores. A clique detection method is used to map the features back onto the binding conformations. The complete algorithm is described in detail, and it is shown that the method can find common superimposition for several test data sets. The method reproduces answers very close to the crystal structure and literature pharmacophores in the examples presented. The basic algorithm is relatively fast and can easily deal with up to 100 compounds and tens of thousands of conformations. The algorithm is also able to handle multiple binding mode problems, which means it can superimpose molecules within the same data set according to two different sets of binding features. We demonstrate the successful use of this algorithm for multiple binding modes for a set of D2 and D4 ligands. PMID- 16711755 TI - Comparative protein modeling of 1-deoxy-D-xylulose-5-phosphate reductoisomerase enzyme from Plasmodium falciparum: a potential target for antimalarial drug discovery. AB - Plasmodium falciparum 1-deoxy-D-xylulose-5-phosphate reductoisomerase (Pf-DXR) is a potential target for antimalarial chemotherapy. The three-dimensional model (3D) of this enzyme was determined by means of comparative modeling through multiple alignment followed by intensive optimization, minimization, and validation. The resulting model demonstrates a reasonable topology as gauged from the Ramachandran plot and acceptable three-dimensional structure compatibility as assessed by the Profiles-3D score. The modeled monomeric subunit consists of three domains: (1) N-terminal NADPH binding domain, (2) connective or linker domain (with most of the active site residues located in this domain), and (3) a C-terminal domain. This structure proved to be consistent with known DXR crystal structures from other species. The predicted active site compared favorably with those of the templates and appears to have an active site with a highly conserved architecture. Additionally, the model explains several site-directed mutagenesis data. Besides using several protein structure-checking programs to validate the model, a set of known inhibitors of DXR were also docked into the active site of the modeled Pf-DXR. The docked scores correlated reasonably well with experimental pIC50 values with a regression coefficient (R2) equal to 0.84. Results of the current study should prove useful in the early design and development of inhibitors by either de novo drug design or virtual screening of large small-molecule databases leading to development of new antimalarial agents. PMID- 16711756 TI - Quantitative structure-activity relationship studies on inhibition of HERG potassium channels. AB - The human ether-a-go-go-related gene (HERG) protein forms the ion channel responsible for the rapidly acting delayed rectifier potassium current, I(Kr), and its blockade is a significant contributor to prolongation of the QT interval. Using descriptors which have clear physicochemical meanings and are familiar to medicinal chemists, we have carried out 2D-quantitative structure-activity relationship (2D-QSAR) studies on 104 HERG channel blockers with diverse structures collected from the literature, and we have formulated interpretable models to guide chemical-modification studies and virtual screening. Statistically significant descriptors were selected by a genetic algorithm, and the final model included the octanol/water partition coefficient, topological polar surface area, diameter, summed surface area of atoms with partial charges from -0.25 to -0.20, and an indicator variable representing the experimental conditions. The statistics were r = 0.839, r2 = 0.704, q2 = 0.671, s = 0.763, and F = 46.6. The correspondence of the molecular determinants derived from the 2D QSAR models with the 3D structural characteristics of the putative binding site in a homology-modeled HERG channel is also discussed. PMID- 16711757 TI - logD7.4 modeling using Bayesian Regularized Neural Networks. Assessment and correction of the errors of prediction. AB - Bayesian Regularized Neural Networks (BRNNs) employing Automatic Relevance Determination (ARD) are used to construct a predictive model for the distribution coefficient logD7.4 from an in-house data set of 5000 compounds with experimental endpoints. A method for assessing the accuracy of prediction is established based upon a query compound's distance to the training set. logD7.4 predictions are also dynamically corrected with an associated library of compounds of continuously updated, experimentally measured logD7.4 values. A comparison of local models and associated libraries comprising separate ionization class subsets of compounds to compounds of a homogeneous ionization class reveals in this case that local models and libraries have no advantage over global models and libraries. PMID- 16711758 TI - Molecular Quantum Similarity Measures from Fermi hole densities: modeling Hammett sigma constants. AB - A new approach, based on the use of fragment Quantum Self-Similarity Measures (MQS-SM) as descriptors of electronic substituent effect in aromatic series, was proposed. The novelty of this approach consists of the fact that the corresponding MQS-SM are not derived, as usual, from ordinary density functions (DF) but from the so-called domain averaged Fermi holes. This approach was applied to the study of substituent effects on the acidobasic dissociation constants in 6 series of para-substituted aromatic carboxylic acids. It has been shown that MQS-SM calculated for each particular set of acids correlate with the Hammett substituent constants. As a consequence, the corresponding similarity measures can be used as new efficient descriptors of the substituent effect, which hopefully could replace empirical sigma constants in QSAR models. PMID- 16711759 TI - A simple approach for indexing the oral druglikeness of a compound: discriminating druglike compounds from nondruglike ones. AB - A knowledge-based simple score has been developed for indexing the oral druglikeness of compounds based on the concept that oral druglikeness should be independent of the drug targets and, thus, are closely related to the global absorption, distribution, metabolism, and excretion related properties. We have considered several simple molecular descriptors as the key determinants of druglikeness. The patterns of the distributions of these molecular descriptors for a set of drug molecules have been extracted using a nonlinear neural network method. We assumed direct correlations of these patterns to the expectation values that a given compound may behave like a drug. On the basis of this assumption, we have defined a simple druglike index or score (DLS) combining the contributions coming from the descriptors considered. This index scales the druglikeness of a compound in the range 0.0-1.0, 1.0 being the highest druglikeness. The index applied for a drug data set, a mixed data set, and three different bioactive databases produced expected features and indicated that even the marketed drugs have druglike scores varying over a considerable range. A total of 73.3% of the drugs considered showed DLS > 0.5, while it is only 44.7% for the HIC-Up compounds (unbiased ligand database). For the ChemBank, Asinex Gold collection, and NCI databases 61.2%, 76.0%, and 79.1% of the compounds have DLS > 0.5. PMID- 16711760 TI - Spatial sign preprocessing: a simple way to impart moderate robustness to multivariate estimators. AB - The spatial sign is a multivariate extension of the concept of sign. Recently multivariate estimators of covariance structures based on spatial signs have been examined by various authors. These new estimators are found to be robust to outlying observations. From a computational point of view, estimators based on spatial sign are very easy to implement as they boil down to a transformation of the data to their spatial signs, from which the classical estimator is then computed. Hence, one can also consider the transformation to spatial signs to be a preprocessing technique, which ensures that the calibration procedure as a whole is robust. In this paper, we examine the special case of spatial sign preprocessing in combination with partial least squares regression as the latter technique is frequently applied in the context of chemical data analysis. In a simulation study, we compare the performance of the spatial sign transformation to nontransformed data as well as to two robust counterparts of partial least squares regression. It turns out that the spatial sign transform is fairly efficient but has some undesirable bias properties. The method is applied to a recently published data set in the field of quantitative structure-activity relationships, where it is seen to perform equally well as the previously described best linear model for these data. PMID- 16711761 TI - Classification tree models for the prediction of blood-brain barrier passage of drugs. AB - The use of classification trees for modeling and predicting the passage of molecules through the blood-brain barrier was evaluated. The models were built and evaluated using a data set of 147 molecules extracted from the literature. In the first step, single classification trees were built and evaluated for their predictive abilities. In the second step, attempts were made to improve the predictive abilities using a set of 150 classification trees in a boosting approach. Two boosting algorithms, discrete and real adaptive boosting, were used and compared. High-predictive classification trees were obtained for the data set used, and the models could be improved with boosting. In the context of this research, discrete adaptive boosting gives slightly better results than real adaptive boosting. PMID- 16711762 TI - 3-D QSAR studies on histone deacetylase inhibitors. A GOLPE/GRID approach on different series of compounds. AB - Docking simulation and three-dimensional quantitative structure-activity relationships (3D-QSARs) analyses were conducted on four series of HDAC inhibitors. The studies were performed using the GRID/GOLPE combination using structure-based alignment. Twelve 3-D QSAR models were derived and discussed. Compared to previous studies on similar inhibitors, the present 3-D QSAR investigation proved to be of higher statistical value, displaying for the best global model r2, q2, and cross-validated SDEP values of 0.94, 0.83, and 0.41, respectively. A comparison of the 3-D QSAR maps with the structural features of the binding site showed good correlation. The results of 3D-QSAR and docking studies validated each other and provided insight into the structural requirements for anti-HDAC activity. To our knowledge this is the first 3-D QSAR application on a broad molecular diversity training set of HDACIs. PMID- 16711763 TI - Development of a chirality-sensitive flexibility descriptor for 3+3D-QSAR. AB - Multidimensional QSAR methodologies can be used to predict the active conformation encoded in the conformational preferences of molecules in an active series by utilizing conformational sampling. In the 3+3D-QSAR approach, the conformational free energy loss is modeled with internal coordinate-based flexibility descriptors. While the pharmacophore point pair distance descriptors introduced earlier proved useful in the construction of QSAR models and in the prediction of important features of the active conformation, they are inherently incapable of describing the chiral arrangement of the pharmacophores. As an improvement, a chirality-sensitive flexibility (CSF) descriptor is now introduced, which is based on the distance between a pharmacophore point and a plane defined by three pharmacophore points. The performance of the CSF descriptor was tested on two active series: 37 endomorphin analogues with opiate activity and 38 PGF2alpha analogues with antinidatory activity. The newly devised descriptor resulted in improved QSAR models in terms of both prediction accuracy and precision of the chiral geometric features of the predicted active conformations. PMID- 16711764 TI - Influence of the conditions in pharmacophore generation, scoring, and 3D database search for chemical feature-based pharmacophore models: one application study of ETA- and ETB-selective antagonists. AB - Using the commercial pharmacophore modeling suite Catalyst, we have studied the influence of the compare.scaledMultiBlobFeatureErrors . Catalyst parameter. The influence of this parameter has been studied in pharmacophore generation, hypothesis scoring, and database searching. This parameter, introduced in Catalyst 4.7, changed its default value in Catalyst 4.8, and it strongly influences the statistical quality of pharmacophore generation, scoring of the hypotheses, and database searching. Two different pharmacophore models have been constructed for the ETA and ETB receptor antagonists. Both models contain one positive ionizable, one negative ionizable, one hydrogen-bond acceptor, one hydrophobic aromatic, and one hydrophobic aliphatic feature. The models have been compared, and some differences in the position of the hydrogen-bond acceptor in the putative binding pocket have been highlighted. PMID- 16711765 TI - Assessing the discriminatory power of scoring functions for virtual screening. AB - The efficiency of scoring functions for hit identification is usually quantified in terms of enrichment factors and enrichment curves. Close inspection of simulated and real score distributions from virtual screening, however, suggests that 'analysis of variance' (ANOVA) is a more reliable method for assessing their performance. Using ANOVA to quantify the discriminatory power of scoring functions with respect to ligands, decoys, and a reproducible reference database has the potential to facilitate the advancement of scoring functions significantly. PMID- 16711766 TI - A genetic-function-approximation-based QSAR model for the affinity of arylpiperazines toward alpha1 adrenoceptors. AB - The genetic function approximation (GFA) algorithm has been used to derive a three-term QSAR equation able to correlate the structural properties of arylpiperazine derivatives with their affinity toward the alpha1 adrenoceptor (alpha1-AR). The number of rotatable bonds, the hydrogen-bond properties, and a variable belonging to a topological family of descriptors (chi) showed significant roles in the binding process toward alpha1-AR. The new model was also compared to a previous pharmacophore for alpha1-AR antagonists and a QSAR model for alpha2-AR antagonists with the aim of finding common or different key determinants influencing both affinity and selectivity toward alpha1- and alpha2 AR. PMID- 16711767 TI - Alignment-free classification of G-protein-coupled receptors using self organizing maps. AB - Proteins are classified mainly on the basis of alignments of amino acid sequences. Drug discovery processes based on pharmacologically important proteins such as G-protein-coupled receptors (GPCRs) may be facilitated if more information is extracted directly from the primary sequences. Here, we investigate an alignment-free approach to protein classification using self organizing maps (SOMs), a kind of artificial neural network, which needs only primary sequences of proteins and determines their relative locations in a two dimensional lattice of neurons through an adaptive process. We first showed that a set of 1397 aligned samples of Class A GPCRs can be classified by our SOM program into 15 conventional categories with 99.2% accuracy. Similarly, a nonaligned raw sequence data set of 4116 samples was categorized into 15 conventional families with 97.8% accuracy in a cross-validation test. Orphan GPCRs were also classified appropriately using the result of the SOM learning. A supposedly diverse family of olfactory receptors formed the most distinctive cluster in the map, whereas amine and peptide families exhibited diffuse distributions. A feature of this kind in the map can be interpreted to reflect hierarchical family composition. Interestingly, some orphan receptors that were categorized as olfactory were somatosensory chemoreceptors. These results suggest the applicability and potential of the SOM program to classification prediction and knowledge discovery from protein sequences. PMID- 16711768 TI - Toward prediction of class II mouse major histocompatibility complex peptide binding affinity: in silico bioinformatic evaluation using partial least squares, a robust multivariate statistical technique. AB - The accurate identification of T-cell epitopes remains a principal goal of bioinformatics within immunology. As the immunogenicity of peptide epitopes is dependent on their binding to major histocompatibility complex (MHC) molecules, the prediction of binding affinity is a prerequisite to the reliable prediction of epitopes. The iterative self-consistent (ISC) partial-least-squares (PLS) based additive method is a recently developed bioinformatic approach for predicting class II peptide-MHC binding affinity. The ISC-PLS method overcomes many of the conceptual difficulties inherent in the prediction of class II peptide-MHC affinity, such as the binding of a mixed population of peptide lengths due to the open-ended class II binding site. The method has applications in both the accurate prediction of class II epitopes and the manipulation of affinity for heteroclitic and competitor peptides. The method is applied here to six class II mouse alleles (I-Ab, I-Ad, I-Ak, I-As, I-Ed, and I-Ek) and included peptides up to 25 amino acids in length. A series of regression equations highlighting the quantitative contributions of individual amino acids at each peptide position was established. The initial model for each allele exhibited only moderate predictivity. Once the set of selected peptide subsequences had converged, the final models exhibited a satisfactory predictive power. Convergence was reached between the 4th and 17th iterations, and the leave-one out cross-validation statistical terms--q2, SEP, and NC--ranged between 0.732 and 0.925, 0.418 and 0.816, and 1 and 6, respectively. The non-cross-validated statistical terms r2 and SEE ranged between 0.98 and 0.995 and 0.089 and 0.180, respectively. The peptides used in this study are available from the AntiJen database (http://www.jenner.ac.uk/AntiJen). The PLS method is available commercially in the SYBYL molecular modeling software package. The resulting models, which can be used for accurate T-cell epitope prediction, will be made freely available online (http://www.jenner.ac.uk/MHCPred). PMID- 16711769 TI - A statistical method for predicting protein unfolding rates from amino acid sequence. AB - The prediction of protein unfolding rates from amino acid sequences is one of the most important challenges in computational biology and chemistry. The analysis on the relationship between protein unfolding rates and physical-chemical, energetic, and conformational properties of amino acid residues provides valuable information to understand and predict the unfolding rates of two- and three-state proteins. We found that the classification of proteins into different structural classes shows an excellent correlation between amino acid properties and unfolding rates of two- and three-state proteins, indicating the importance of native-state topology in determining the protein unfolding rates. We have formulated three independent linear regression equations to different structural classes of proteins for predicting their unfolding rates from amino acid sequences and obtained an excellent agreement between predicted and experimentally observed unfolding rates of proteins; the correlation coefficients are 0.999, 0.990, and 0.992, respectively, for all-alpha, all-beta, and mixed class proteins. Further, we have derived a general equation applicable to all structural classes of proteins, which can be used for predicting the unfolding rates for proteins of an unknown structural class. We observed a correlation of 0.987 and 0.930, respectively, for back-check and jack-knife tests. These accuracy levels are better than those of other methods in the literature. PMID- 16711770 TI - In silico analysis of the thermodynamic stability changes of psychrophilic and mesophilic alpha-amylases upon exhaustive single-site mutations. AB - Identifying sequence modifications that distinguish psychrophilic from mesophilic proteins is important for designing enzymes with different thermodynamic stabilities and to understand the underlying mechanisms. The PoPMuSiC algorithm is used to introduce, in silico, all the single-site mutations in four mesophilic and one psychrophilic chloride-dependent alpha-amylases and to evaluate the changes in thermodynamic stability. The analysis of the distribution of the sequence positions that could be stabilized upon mutation shows a clear difference between the three domains of psychrophilic and mesophilic alpha amylases. Most of the mutations stabilizing the psychrophilic enzyme are found in domains B and C, contrary to the mesophilic proteins where they are preferentially situated in the catalytic domain A. Moreover, the calculations show that the environment of some residues responsible for the activity of the psychrophilic protein has evolved to reinforce favorable interactions with these residues. In the second part, these results are exploited to propose rationally designed mutations that are predicted to confer to the psychrophilic enzyme mesophilic-like thermodynamic properties. Interestingly, most of the mutations found in domain C strengthen the interactions with domain A, in agreement with suggestions made on the basis of structural analyses. Although this study focuses on single-site mutations, the thermodynamic effects of the recommended mutations should be additive if the mutated residues are not close in space. PMID- 16711771 TI - CRAACK: consensus program for NMR amino acid type assignment. AB - Protein peak spectrum assignment is a prerequisite of the nuclear magnetic resonance study of a molecule. We present here a computer tool which proposes the determination of the amino acid type from the values of the chemical shifts. This tool is based on two consensus algorithms based on several published typing algorithms and was trained and extensively tested against the Biological Magnetic Resonance Bank chemical shift data bank. The first one accomplishes the analysis with support vector machine technology, grouping related amino acids together, and presents a mean rate of success above 90% on the test set. The second one uses a classical consensus algorithm of vote. Furthermore, secondary structural prediction is available. This tool can be used for assisting manual assignment of peptides and proteins and can also be used as a step in an automated approach to assignment. This program has been called CRAACK and is publicly available at the following URL: http://abcis.cbs.cnrs.fr/craack. PMID- 16711772 TI - Diagnostic tools to determine the quality of "transparent" regression-based QSARs: the "modelling power" plot. AB - A bivariate plot is presented for comparing two or more QSAR models. It is based on two new statistics associated with a regression model, the "descriptive power" (Dp), which is estimated through the relative uncertainty of model coefficients, and the "predictive power" (Pp), which is estimated through both the fitted and cross-validated explained variance of the response variable (i.e., biological activity). An algorithm was developed for performing equivalent multiple linear regression and partial-least-squares calculations. The results were validated by comparison with widely accepted commercial software. Dp and Pp statistics are defined to vary from 0 to 100%, so the modeler has a intuitive impression of the descriptive (i.e., global importance of the selected descriptors) and predictive (i.e., possibility of performing QSAR or just SAR estimations) power. These statistics represent a point in the Dp versus Pp "modelling power" plot, which facilitates visual multiple models comparison, but also could be used to substitute classical statistics and could even be combined to obtain a unique parameter to define (or compare) the model's quality. PMID- 16711773 TI - Scaling of ballistic deposition from a Langevin equation. AB - An exact lattice Langevin equation is derived for the ballistic deposition model of surface growth. The continuum limit of this equation is dominated by the Kardar-Parisi-Zhang (KPZ) equation at all length and time scales. For a one dimensional substrate the solution of the exact lattice Langevin equation yields the KPZ scaling exponents without any extrapolation. For a two-dimensional substrate the scaling exponents are different from those found from computer simulations. This discrepancy is discussed in relation to analytic approaches to the KPZ equation in higher dimensions. PMID- 16711774 TI - Glass transition of glycerol in the volume-temperature plane. AB - We assess the relative importance of spatial congestion and lowered temperature in the slowing dynamics of supercooled glycerol near the glass transition. We independently vary both volume V and temperature T by applying high pressure and monitor the dynamics by measuring the dielectric susceptibility. Our results demonstrate that both variables are control variables of comparable importance. However, a generalization of the concept of fragility of a glass-former shows that the dynamics are quantitatively more sensitive to fractional changes in V than T. We identify a connection between the fragility and a recently proposed density-temperature scaling which indicates that this conclusion holds for other liquids and polymers. PMID- 16711775 TI - Entropy-based measure of structural order in water. AB - We analyze the nature of the structural order established in liquid TIP4P water in the framework provided by the multiparticle correlation expansion of the statistical entropy. Different regimes are mapped onto the phase diagram of the model upon resolving the pair entropy into its translational and orientational components. These parameters are used to quantify the relative amounts of positional and angular order in a given thermodynamic state, thus allowing a structurally unbiased definition of low-density and high-density water. As a result, the structurally anomalous region--within which both types of order are simultaneously disrupted by an increase of pressure at constant temperature--is clearly identified through extensive molecular-dynamics simulations. PMID- 16711776 TI - Defect-mediated stripe reordering in wrinkles upon gradual changes in compression direction. AB - We study the stripe rearrangements in microwrinkles subjected to gradual changes in the compression direction. The spatially averaged stripe orientation follows the changing compression direction with a deviation of 3-4 degrees. Meanwhile, the local stripe orientation stays constant for a certain period and a dislocation gliding nearby causes it to jump suddenly in the direction of compression. This sequence repeatedly occurs over the surface. The regular pattern transiently appears in the stripe orientation as a result of the organized glide motions, indicating the interaction between dislocations. PMID- 16711777 TI - Electrically tuned phase transition and band structure in a liquid-crystal infilled photonic crystal. AB - We studied a nematic liquid crystal (LC) cylinder under the action of an axial electric field E(0). Elaborate modeling of the free energy leads to the conclusion that the configuration of the molecules is "escaped radial" for low E(0); a phase transition, however, occurs for a critical value E(c), the configuration becoming axial for E(0) >E(c). From these results, the position dependent dielectric tensor is determined and the photonic band (PB) structure is calculated for a photonic crystal of LC cylinders. It is shown that by varying E(0) a PB gap can be fully tuned from open to closed. Also, switching to a supercritical field can give rise to interesting polarization and directional effects in the propagation of light. PMID- 16711778 TI - Anomalously slow domain growth in fluid membranes with asymmetric transbilayer lipid distribution. AB - The effect of asymmetry in the transbilayer lipid distribution on the dynamics of phase separation in fluid vesicles is investigated numerically. This asymmetry is shown to set a spontaneous curvature for the domains that alter the morphology and dynamics considerably. For moderate tension, the domains are capped and the spontaneous curvature leads to anomalously slow dynamics, as compared to the case of symmetric bilayers. In contrast, in the limiting cases of high and low tensions, the dynamics proceeds toward full phase separation. PMID- 16711779 TI - Short-time dynamics of the helix-coil transition in polypeptides. AB - We study the critical relaxation of the helix-coil transition in all-atom models of polyalanine chains. We show that at the critical temperature the decay of a completely helical conformation can be described by scaling relations that allow us estimating the pertinent critical exponents. The present approach opens a new way for characterizing transitions in proteins and may lead to a better understanding of their folding mechanism. An application of the technique to the 34-residue human parathyroid fragment PTH(1-34) supports universality of the helix-coil transition in homopolymers and (helical) proteins. PMID- 16711780 TI - Fluctuations and correlations in lattice models for predator-prey interaction. AB - Including spatial structure and stochastic noise invalidates the classical Lotka Volterra picture of stable regular population cycles emerging in models for predator-prey interactions. Growth-limiting terms for the prey induce a continuous extinction threshold for the predator population whose critical properties are in the directed percolation universality class. We discuss the robustness of this scenario by considering an ecologically inspired stochastic lattice predator-prey model variant where the predation process includes next nearest-neighbor interactions. We find that the corresponding stochastic model reproduces the above scenario in dimensions 1< d < or =4, in contrast with the mean-field theory, which predicts a first-order phase transition. However, the mean-field features are recovered upon allowing for nearest-neighbor particle exchange processes, provided these are sufficiently fast. PMID- 16711781 TI - Multiplicative noise in the longitudinal mode dynamics of a bulk semiconductor laser. AB - We analyze theoretically and experimentally the influence of current noise on the longitudinal mode hopping dynamics of a bulk semiconductor laser. It is shown that the mean residence times on each mode have different sensitivity to external noise added to the bias current. In particular, an increase of the noise level enhances the residence time on the longitudinal mode that dominates at low current, evidencing the multiplicative nature of the stochastic process. A two mode rate equation model for a semiconductor laser is able to reproduce the experimental findings. Under a suitable separation of the involved time scales, the model can be reduced to a one-dimensional bistable potential system with a multiplicative stochastic term related to the current noise strength. The reduced model clarifies the influence of the different noise sources on the hopping dynamics. PMID- 16711782 TI - Analytical approach to sorting in periodic and random potentials. AB - There has been a recent revolution in the ability to manipulate micrometer-sized objects on surfaces patterned by traps or obstacles of controllable configurations and shapes. One application of this technology is to separate particles driven across such a surface by an external force according to some particle characteristic such as size or index of refraction. The surface features cause the trajectories of particles driven across the surface to deviate from the direction of the force by an amount that depends on the particular characteristic, thus leading to sorting. While models of this behavior have provided a good understanding of these observations, the solutions have so far been primarily numerical. In this paper we provide analytic predictions for the dependence of the angle between the direction of motion and the external force on a number of model parameters for periodic as well as random surfaces. We test these predictions against exact numerical simulations. PMID- 16711783 TI - Single-species three-particle reactions in one dimension. AB - Renormalization group calculations for fluctuation-dominated reaction-diffusion systems are generally in agreement with simulations and exact solutions. However, simulations of the single-species reactions 3A -->(O,A,2A) at their upper critical dimension d(c)=1 have found asymptotic densities argued to be inconsistent with renormalization group predictions. We show that this discrepancy is resolved by inclusion of the leading corrections to scaling, which we derive explicitly and show to be universal, a property not shared by the A+A- >(O,A) reactions. Finally, we demonstrate that two previous Smoluchowski approaches to this problem reduce, with various corrections, to a single theory which surprisingly yields the same asymptotic density as the renormalization group. PMID- 16711784 TI - Effect of a slit-shaped trap on depletion kinetics within a microchannel. AB - The diffusion-limited trapping reaction kinetics of the growth of the depletion zone within and around a "slit-shaped" trap in a flat microchannel was studied experimentally and numerically. In the experiment, an ellipse-shaped laser beam acted as a slit trap in a long, flat capillary, and the trapping reaction is photobleaching of fluorescein dye. The parameter studied was the theta distance, i.e., the distance from the trap to the point where the reactant concentration has been locally depleted to the specific survival fraction [theta] of its initial bulk value. When the trap is perfect, then, due to the geometry of the trap and the reactor, as many as three time regimes can be found, with up to two crossover transitions. The number of crossovers is determined by the relative sizes of the trap and the microreactor. In the case of two crossovers, we show that the first crossover relates to the length of the trap, while the second crossover relates to the width of the reactor. When the slit trap is imperfect and its width cannot be neglected, as is the case in the experiments, a nontrivial early behavior is observed, followed by two regions in time, separated by a single crossover only. PMID- 16711786 TI - Evaporation-induced flow in an inviscid liquid line at any contact angle. AB - The problem of potential flow inside an evaporating liquid line, shaped as an infinitely long cylindrical segment lying on a flat surface, is considered and an analytical solution is obtained for any contact angle in (0, pi). In this way, microflow details inside linear liquid bodies evaporating on hydrophilic, hydrophobic, and strongly hydrophobic substrates can now be obtained. The mathematical formulation employs the velocity potential and stream function formulations in bipolar coordinates and the solution is obtained using the technique of Fourier transform. Both pinned and depinned contact lines are considered. The solution is applicable to any evaporation mechanism but for illustration purposes numerical results are presented here for the particular case of kinetically controlled evaporation. For hydrophilic substrates, the flow inside the evaporating liquid line is directed towards the edges for pinned contact lines, thus, promoting a coffee stain effect. The opposite flow direction is observed for depinned contact lines. However, for strongly hydrophobic substrates, flow is directed outwards for both pinned and depinned contact lines, but owing to its low magnitude compared to that on hydrophilic substrates, a craterlike colloidal deposit should be expected rather than a ringlike deposit, in agreement with experimental observations. PMID- 16711785 TI - Hamiltonian theory of stochastic acceleration. AB - Stochastic acceleration, defined in terms of a stochastic equation of motion for the acceleration, is derived from a Hamiltonian model. A free particle is coupled bilinearly to a harmonic bath through the particle's momentum and coordinate. Under appropriate conditions, momentum coupling induces velocity diffusion which is not destroyed by the spatial coupling. Spatial-momentum coupling may induce spatial subdiffusion. The thermodynamic equilibrium theory presented in this paper does not violate the second law of thermodynamics, although the average velocity squared of the particle may increase in time without bound. PMID- 16711787 TI - Diffusion of a granular pulse in a rotating drum. AB - The diffusion of a pulse of small grains in an horizontal rotating drum is studied through discrete elements methods simulations. We present a theoretical analysis of the diffusion process in a one-dimensional confined space in order to elucidate the effect of the confining end-plate of the drum. We then show that the diffusion is neither subdiffusive nor superdiffusive but normal. This is demonstrated by rescaling the concentration profiles obtained at various stages and by studying the time evolution of the mean squared deviation. Finally we study the self-diffusion of both large and small grains, and we show that it is normal and that the diffusion coefficient is independent of the grain size. PMID- 16711788 TI - Shock wave propagation in vibrofluidized granular materials. AB - Shock wave formation and propagation in vertically vibrated quasi-two-dimensional granular materials are studied by digital high speed photography. Steep density and temperature wave fronts form at the bottom of the granular layer when the layer collides with vibrating plate. Then the fronts propagate upwards through the layer. The temperature front is always in the transition region between the upward and downward granular flows. The effects of driving parameters and particle number on the shock are also explored. PMID- 16711789 TI - Dynamics of axial segregation in granular slurries: parallel experiments and influence of aspect ratio and periodic tilting. AB - An efficient technique for conducting rotating tumbler experiments in parallel is introduced and used to study the effect of tumbler length and periodic tilting of the tumbler on axial segregation. When rotated, bidisperse granular slurries segregate into what appear at the surface to be alternating bands of larger and smaller particles. The number of bands increases linearly with tumbler length while the fractional area occupied by each type of band is constant. Periodic tilting of the rotation axis induces a periodic axial flow of particles in the flowing layer. For the range of tilt angle amplitudes investigated (0 degrees 3.5 degrees), the number of bands decreases with increasing angle, but the rate of merging and the fractional area of bands rich in smaller particles are unaffected. PMID- 16711790 TI - Structural signatures of the unjamming transition at zero temperature. AB - We study the pair correlation function g(r) for zero-temperature, disordered, soft-sphere packings just above the onset of jamming. We find distinct signatures of the transition in both the first and split second peaks of this function. As the transition is approached from the jammed side (at higher packing fraction) the first peak diverges and narrows on the small-r side to a delta function. On the high-r side of this peak, g(r) decays as a power law. In the split second peak, the two subpeaks are both singular at the transition, with power-law behavior on their low-r sides and step-function drop-offs on their high-r sides. These singularities at the transition are reminiscent of empirical criteria that have previously been used to distinguish glassy structures from liquid ones. PMID- 16711791 TI - Experimental evidence of shock mitigation in a Hertzian tapered chain. AB - We present an experimental study of the mechanical impulse propagation through a horizontal alignment of elastic spheres of progressively decreasing diameter phi(n): namely, a tapered chain. Experimentally, the diameters of spheres which interact via the Hertz potential are selected to keep as close as possible to an exponential decrease, phi(n+1) = (1-q)phi(n), where the experimental tapering factor is either q(1) approximately equal to 5.60% or q(2) approximately equal to 8.27%. In agreement with recent numerical results, an impulse initiated in a monodisperse chain (a chain of identical beads) propagates without shape changes and progressively transfers its energy and momentum to a propagating tail when it further travels in a tapered chain. As a result, the front pulse of this wave decreases in amplitude and accelerates. Both effects are satisfactorily described by the hard-sphere approximation, and basically, the shock mitigation is due to partial transmissions, from one bead to the next, of momentum and energy of the front pulse. In addition when small dissipation is included, better agreement with experiments is found. A close analysis of the loading part of the experimental pulses demonstrates that the front wave adopts a self-similar solution as it propagates in the tapered chain. Finally, our results corroborate the capability of these chains to thermalize propagating impulses and thereby act as shock absorbing devices. PMID- 16711792 TI - Dynamic self-assembly of magnetic particles on the fluid interface: surface-wave mediated effective magnetic exchange. AB - We report on studies of dynamic self-assembled structures induced by a vertical alternating magnetic field in an ensemble of magnetic particles suspended on a liquid surface. We find the formation of nontrivially ordered dynamic snakelike objects in a certain range of field magnitudes and frequencies. In order to probe the properties of the "snakes," we study their magnetic response to in-plane magnetic field applied at different angles with respect to its axis. The segments of the snake exhibit long-range antiferromagnetic ordering mediated by the surface waves, while each segment is composed of ferromagnetically aligned chains of microparticles. We propose a simple phenomenological model where the effect of surface waves is replaced by an effective exchange interaction to describe the observations. In the framework of the proposed model, the effective exchange constant corresponding to different regimes of magnetic driving was extracted from the experimental data. PMID- 16711793 TI - Rearrangements in a two-dimensional packing of disks. AB - Several aspects of the dynamics of a granular two-dimensional (2D) packing of disks slowly tilted until the system loses stability and an avalanche takes place are discussed. The evolution of the system, constructed with monodisperse disks placed on a thin cell, is studied by image analysis. As in the 3D case (packing of spheres), the system undergoes several rearrangements of different magnitude before the avalanche takes place. For thick systems, not only are small rearrangements detected but also displacements of large clusters of disks are observed in the bulk and on the free surface of the packing. In particular, characteristic angles and the avalanche mass were determined for samples of different heights. On thick systems, velocity fields of large rearrangements are presented and changes in the internal structure of the packing produced by these rearrangements are analyzed. It is found that the main effects of rearrangements is to increase the disorder of the system. Also, as the disorder of the system increases its stability threshold decreases. PMID- 16711794 TI - Devitrification of colloidal glasses in real space. AB - Confocal scanning laser microscopy has been used to quantitatively analyze the structure and dynamics of concentrated suspensions of spherical colloids in which the magnitude of the short-range attractive potential is increased by adding nonadsorbing polymers. These systems undergo a reentrant glass transition upon increasing polymer concentration. We find that melting of the glass is accompanied by significant changes in the displacement distribution and its moments. However, no significant variations have been detected in the shapes of the displacement distributions. Moreover, structural correlation functions and the magnitude of local density fluctuations do not vary significantly between the glass states and the fluid. Considering our experimental setup, these observations imply that local density fluctuations cannot be larger than a few percent of the average density. PMID- 16711795 TI - Magnetic-field-induced orientational order in the isotropic phase of hard colloidal platelets. AB - The magnetic-field-induced orientational order in the isotropic phase of colloidal gibbsite [Al(OH)3] platelets is studied by means of optical birefringence and small-angle x-ray scattering (SAXS) techniques. The suspensions display field-induced ordering at moderate field strengths (a few Tesla), which increases with increasing particle concentration. The gibbsite particles align their normals perpendicular to the magnetic field and hence possess a negative anisotropy of their diamagnetic susceptibility Delta(chi). The results can be described following a simple, Onsager-like approach. A simplified model is derived that allows one to obtain the orientational distribution function directly from the scattering data. However, it leads to an underestimate of the diamagnetic susceptibility anisotropy Delta(chi). This accounts for the difference between the Delta(chi) values provided by the two experimental techniques (SAXS and magneto-optics). The order of magnitude Delta(chi) approximately 10(-22) J/T(2) lies in between that of goethite suspensions and that of suspensions of organic particles. PMID- 16711796 TI - Growth of fractal electrodeposited aggregates under action of electric and magnetic fields using a modified diffusion-limited aggregation algorithm. AB - We have simulated the two-dimensional growth of fractal aggregates produced in electrodeposition experiments with axial symmetry in the presence of electric and magnetic fields. A modified diffusion-limited aggregation algorithm based on the Monte Carlo method is used in order to simulate cluster growth under the action of Coulomb and Lorentz forces, taking into account the thermal energy. The ion particle movement has each step biased by the resultant force; in the algorithm, it is mediated by the Boltzmann term. The electric voltage between the electrodes tends to compact the aggregates and reduce the effect of screening. The Lorentz force provides a spiral form for aggregates which twists according to the magnetic field direction and intensity. A function was defined to measure the chirality of the system. The fractal dimension was also calculated to measure the influence of the electric and magnetic fields as well as the temperature during the growth process. Good agreement with experimental results was observed. PMID- 16711797 TI - Effect of many-body interactions on the bulk and interfacial phase behavior of a model colloid-polymer mixture. AB - We study a model suspension of sterically stabilized colloidal particles and nonadsorbing ideal polymer coils, both in bulk and adsorbed against a planar hard wall. By integrating out the degrees of freedom of the polymer coils, we derive a formal expression for the effective one-component Hamiltonian of the colloids. We employ an efficient Monte Carlo simulation scheme for this mixture based on the exact effective colloid Hamiltonian; i.e., it incorporates all many-body interactions. The many-body character of the polymer-mediated effective interactions between the colloids yields bulk phase behavior and adsorption phenomena that differ substantially from those found for pairwise simple fluids. We determine the phase behavior for size ratios q=sigma(p)/sigma(c)=1, 0.6, and 0.1, where sigma(c) and sigma(p) denote the diameters of the colloids and polymer coils, respectively. For q=1 and 0.6, we find both a fluid-solid and a stable colloidal gas-liquid transition with an anomalously large bulk liquid regime caused by the many-body interactions. We compare the phase diagrams obtained from simulations with the results of the free-volume approach and with direct simulations of the true binary mixture. Although we did not simulate the polymer coils explicitly, we are able to obtain the three partial structure factors and radial distribution functions. We compare our results with those obtained from density functional theory and the Percus-Yevick approximation. We find good agreement between all results for the structure. We also study the mixture in contact with a single hard wall for q=1. Upon approach of the gas-liquid binodal, we find far from the triple point, three layering transitions in the partial wetting regime. PMID- 16711798 TI - Microrheology and structure of a yield-stress polymer gel. AB - The small-scale rheology of Carbopol ETD 2050, a polymer gel with a yield stress, is studied as a function of polymer concentration by measuring the diffusion of submicron-sized spherical fluorescent particles suspended in gel. Dynamic light scattering is used to determine the mean-squared displacement (of the particles as a function of lag time t. Fluorescence microscopy is used to track the particle trajectories directly, from which and the van Hove correlation function are determined. From our results we calculate the microrheological viscous and elastic moduli of the material. The two techniques cover complementary ranges of tau and and give results that agree well. The microrheological moduli are substantially smaller than the bulk values as determined by conventional shear rheometry. The bulk viscoelastic behavior is dominated by the elastic modulus, while at low enough concentrations and high enough frequencies the microrheological response is predominantly viscous. These results will be discussed in the context of the gel structure. PMID- 16711799 TI - Orientation dynamics of weakly Brownian particles in periodic viscous flows. AB - Evolution equations for the orientation distribution of axisymmetric particles in periodic flows are derived in the regime of small but nonzero Brownian rotations. The equations are based on a multiple time scale approach that allows fast computation of the relaxation processes leading to statistical equilibrium. The approach has been applied to the calculation of the effective viscosity of a thin disk suspension in an oscillating strain flow. PMID- 16711800 TI - Phase separation in charge-stabilized colloidal suspensions: influence of nonlinear screening. AB - The phase behavior of charge-stabilized colloidal suspensions is modeled by a combination of response theory for electrostatic interparticle interactions and variational theory for free energies. Integrating out degrees of freedom of the microions (counterions, salt ions), the macroion-microion mixture is mapped onto a one-component system governed by effective macroion interactions. Linear response of microions to the electrostatic potential of the macroions results in a screened-Coulomb (Yukawa) effective pair potential and a one-body volume energy, while nonlinear response modifies the effective interactions [A. R. Denton, Phys. Rev. E 70, 031404 (2004)]. The volume energy and effective pair potential are taken as input to a variational free energy, based on thermodynamic perturbation theory. For both linear and first-order nonlinear effective interactions, a coexistence analysis applied to aqueous suspensions of highly charged macroions and monovalent microions yields bulk separation of macroion rich and macroion-poor phases below a critical salt concentration, in qualitative agreement with predictions of related linearized theories [R. van Roij, M. Dijkstra, and J.-P. Hansen, Phys. Rev. E 59, 2010 (1999); P. B. Warren, J. Chem. Phys. 112, 4683 (2000)]. It is concluded that nonlinear screening can modify phase behavior but does not necessarily suppress bulk phase separation of de ionized suspensions. PMID- 16711801 TI - Phase behavior of mixtures of oppositely charged protein nanoparticles at asymmetric charge ratios. AB - We present experimental and theoretical results for the phase behavior of mixtures of oppositely charged globular protein molecules in aqueous solutions containing monovalent salt. These colloidal mixtures are interesting model systems, on the one hand for electrolyte solutions ("colloidal ionic liquids"), and on the other for mixtures of oppositely charged (bio)macromolecules, colloids, micelles, etc., with the range of the electrostatic interactions (Debye length) easily tunable from much smaller to much larger than the particle size, simply by adding different amounts of monovalent salt. In this paper we investigate the phase behavior of such mixtures in the case that equally sized colloids have a large difference in charge magnitude. This is possible at any mixing ratio because small ions compensate any colloidal charge asymmetry. Our experimental system is based on lysozyme, a positively charged "hard" globular protein molecule, and succinylated lysozyme, a chemical modification of lysozyme which is negatively charged. By changing the solution pH we can adjust the ratio of charge between the two molecules. To describe phase separation into a dilute phase and a dense "complex" phase, a thermodynamic model is set up in which we combine the Carnahan-Starling-van der Waals equation of state with a heterogeneous Poisson-Boltzmann cell model and include the possibility that protein molecules adjust their charge when they move from one phase to the other (charge regulation). The theory uses the nonelectrostatic attraction strength as the only adjustable parameter and reasonably well reproduces the data in that complexation is only possible at intermediate , not too asymmetric mixing ratios, and low enough ionic strength and temperature. PMID- 16711802 TI - Colloid interaction and pair correlation function of one-dimensional colloid polymer systems. AB - The interaction and pair correlation function of weakly charged colloidal particles in quasi-one-dimensional colloid-polymer systems are determined by enhanced video microscopy and digital image analysis. The pair correlation function is found to depend not only on the polymer concentration, but also on the degree of confinement; in particular, it depends on whether the channel width is such that mutual passage of the colloid particles is possible or not. These findings are compared with exact results on short-range order in linear continuous systems. PMID- 16711803 TI - Discrete elastic model for two-dimensional melting. AB - We present a network model for the study of melting and liquid structure in two dimensions, the first in which the presence and energy of topological defects (dislocations and disclinations) and of geometrical defects (elemental voids) can be independently controlled. Interparticle interaction is via harmonic springs and control is achieved by Monte Carlo moves which springs can either be orientationally "flipped" between particles to generate topological defects, or can be "popped" in force-free shape, to generate geometrical defects. With the geometrical defects suppressed the transition to the liquid phase occurs via disclination unbinding, as described by the Kosterlitz-Thouless-Halperin-Nelson Young model and found in soft potential two-dimensional (2D) systems, such as the dipole-dipole potential [H. H. von Grunberg, Phys. Rev. Lett. 93, 255703 (2004)]. By contrast, with topological defects suppressed, a disordering transition, the Glaser-Clark condensation of geometrical defects [M. A. Glaser and N. A. Clark, Adv. Chem. Phys. 83, 543 (1993); M. A. Glaser, (Springer-Verlag, Berlin, 1990), Vol. 52, p. 141], produces a state that accurately characterizes the local liquid structure and first-order melting observed in hard-potential 2D systems, such as hard disk and the Weeks-Chandler-Andersen (WCA) potentials (M. A. Glaser and co workers, see above). Thus both the geometrical and topological defect systems play a role in melting. The present work introduces a system in which the relative roles of topological and geometrical defects and their interactions can be explored. We perform Monte Carlo simulations of this model in the isobaric isothermal ensemble, and present the phase diagram as well as various thermodynamic, statistical, and structural quantities as a function of the relative populations of geometrical and topological defects. The model exhibits a rich phase behavior including hexagonal and square crystals, expanded crystal, dodecagonal quasicrystal, and isotropic liquid phases. In this system the geometrical defects effectively control the melting, reducing the solid-liquid transition temperature by a factor of relative to the topological-only case. The local structure of the dense liquid has been investigated and the results are compared to that from simulations of WCA systems. PMID- 16711804 TI - Derivative-based analysis for temperature and pressure evolution of dielectric relaxation times in vitrifying liquids. AB - The derivative-based analysis for detecting regions of the validity of the Vogel Fulcher-Tammann (VFT) dependence for superpressed and supercooled liquids is discussed. For the temperature (T) path the analysis introduced by Stickel [J. Chem. Phys. 104, 2043 (1996); J. Chem. Phys. 107, 1086 (1997)] is recalled. For the pressure (P) path the derivation based on the counterpart of the VFT dependence proposed in Paluch [J. Phys.: Condens. Mater 10, 4131 (1998)] is presented. The appearance of two ideal glass temperatures (T(0)) or pressures (P(0)), fragility strength coefficients (D(T), D(p)), and prefactors [formula- see text] for VFT equations in following dynamical domains, i.e., high temperature (DeltaT(high)) and low-temperature (DeltaT(low)) or low-pressure (DeltaP(low)), and high-pressure (DeltaP(high)), is stressed. It is noteworthy that the values of T(0)(DeltaT(high)) > T(0)(DeltaT(low)), D(T)(DeltaT(high)) << D(T)(DeltaT(low)), and [formula--see text]. Analogous behavior was noted for isothermal DeltaP(L) and DeltaP(H) dynamic domains. A similar derivative-based approach is also applied to test the validity of the mode coupling theory (MCT) critical-like equation [formula--see text]. It yields the temperature T(X) and the MCT power ("critical") exponent [formula--see text] exclusively from the simple linear regression. The extension of such an analysis for the pressure path is also given. The hardly discussed question of the error of estimations of [formula--see text] and T(X) is emphasized. The relation between the derivative based behavior mentioned above and the apparent activation enthalpy (temperature path) or the apparent activation volume (pressure path) is indicated. The presented analysis was applied to discuss the dynamic crossovers in supercooled and superpressed diethyl phthalate, based on experimental data supplemented by those given in Pawlus [Phys. Rev. E 68, 021503 (2003)]. PMID- 16711805 TI - Structural explanation of the rheology of a colloidal suspension under high dc electric fields. AB - In this work we describe the electrorheology of suspensions consisting of hematite (alpha-Fe2O3) particles dispersed in silicone oil in the presence of large dc electric fields. If an electric field pulse is applied to the systems, it is possible to estimate the time that the electrorheological (ER) fluid takes to reach its final microstructure in the presence of the field. Our results indicate that response times of several seconds are typical, and that this time decreases with the field strength. Conventional shear-rate sweeps indicate the existence of a well-defined dynamic yield stress and a shear-thinning behavior. Interestingly, both the yield stress and the shear-thinning slope alpha [relating the viscosity, eta, and the shear rate, .gamma, as eta=alphagamma(.-b) + eta (infinity)] show a linear dependence on the field strength, E, in disagreement with the E2 dependence often reported. This deviation is associated with changes in the conductivity of the dispersion medium with the field strength. A simple calculation of the interactions present in our ER fluid demonstrates that the ER behavior is entirely controlled by hydrodynamic (proportional to .gamma) and electrical forces (proportional to E). This is confirmed by the collapse of all experimental results in a single master curve when the relative viscosity is plotted against the ratio .gamma/E. Careful attention has been paid in this work to the microstructure of the suspensions in the presence of both shear and electric fields simultaneously: the particles gather themselves on the walls of the electrorheological measurement cell, forming aggregates with cylindrical symmetry, shaped as rings or lamellas of solids. The electric field induced increase in viscosity is the consequence of the balance between two actions: that of the electric field, tending to keep particles together, and that of the shear field, forcing the flow of the liquid phase in the regions between rings or between rings and walls. PMID- 16711806 TI - Dynamics and thermodynamics of the glass transition. AB - The principal theme of this paper is that anomalously slow, super-Arrhenius relaxations in glassy materials may be activated processes involving chains of molecular displacements. As pointed out in a preceding paper with Lemaitre, the entropy of critically long excitation chains can enable them to grow without bound, thus activating stable thermal fluctuations in the local density or molecular coordination of the material. I argue here that the intrinsic molecular scale disorder in a glass plays an essential role in determining the activation rate for such chains, and show that a simple disorder-related correction to the earlier theory recovers the Vogel-Fulcher law in three dimensions. A key feature of this theory is that the spatial extent of critically long excitation chains diverges at the Vogel-Fulcher temperature. I speculate that this diverging length scale implies that, as the temperature decreases, increasingly large regions of the system become frozen and do not contribute to the configurational entropy, and thus ergodicity is partially broken in the super-Arrhenius region above the Kauzmann temperature T(K). This partially broken ergodicity seems to explain the vanishing entropy at T(K) and other observed relations between dynamics and thermodynamics at the glass transition. PMID- 16711807 TI - Molecular dynamics study of orientational cooperativity in water. AB - Recent experiments on liquid water show collective dipole orientation fluctuations dramatically slower than expected (with relaxation time > 50 ns) [D.P. Shelton, Phys. Rev. B 72, 020201(R) (2005)]. Molecular dynamics simulations of extended simple point charge (SPC/E) water show a large vortexlike structure of the dipole field at ambient conditions surviving over [J. Higo, Proc. Natl. Acad. Sci. U.S.A. 98, 5961 (2001)]. Both results disagree with previous results on water dipoles in similar conditions, for which autocorrelation times are a few picoseconds. Motivated by these recent results, we study the water dipole reorientation using molecular dynamics simulations of the SPC/E model in bulk water for temperatures ranging from ambient 300 K down to the deep supercooled region of the phase diagram at 210 K. First, we calculate the dipole autocorrelation function and find that our simulations are well described by a stretched exponential decay, from which we calculate the orientational autocorrelation time t(alpha). Second, we define a second characteristic time, namely, the time required for the randomization of molecular dipole orientation, the self-dipole randomization time t(r), which is an upper limit on t(alpha); we find that t(r) is approximately equal to 5t(alpha). Third, to check if there are correlated domains of dipoles in water which have large relaxation times compared to the individual dipoles, we calculate the randomization time t(box) of the site dipole field, the net dipole moment formed by a set of molecules belonging to a box of edge L(box). We find that the site-dipole randomization time t(box) is approximately equal to 2.5t(alpha) for L(box) approximately equal to 3 A, i.e., it is shorter than the same quantity calculated for the self-dipole. Finally, we find that the orientational correlation length is short even at low T. PMID- 16711808 TI - Effects of oil on the curvature elastic properties of nonionic surfactant films: thermodynamics of balanced microemulsions. AB - The free energy of nonionic balanced microemulsions based on nonionic surfactants are analyzed using experimental data from (i) phase behavior, (ii) osmotic compressibility of the balanced microemulsion structure, which is obtained from small angle neutron scattering (SANS) experiments, and (iii) data on interfacial tensions obtained by T. Sottmann and R. Strey [J. Chem. Phys. 106 8606 (1997)]. The balanced microemulsion, where the spontaneous curvature vanishes at equal volumes of water and oil, has a finite swelling with the solvent with a minimum surfactant volume fraction, Phi*(S). At higher surfactant concentrations the balanced microemulsion phase having the surfactant volume fraction Phi(S1) coexists with a lamellar phase of volume fraction Phi(S2). Under the constraint of Phi(W) = Phi(O), where Phi(W) and Phi(O) are the water and oil volume fractions, respectively, the free energy density can be written as an expansion in the surfactant concentration. While the phase equilibria only depend on relative values of the expansion coefficients, absolute values can be obtained from compressibility and interfacial tension data. The osmotic compressibility of the surfactant film was measured by SANS through contrast matching water and oil. The phase behavior of nonionic surfactant-water-oil systems depends strongly on the chain length of the oil, when comparing a homologous series from octane to hexadecane using the same surfactant, here being pentaethylene oxide dodecyl ether (C(12)E(5)). The three concentrations Phi*(S), Phi(S1), and Phi(S2) increase markedly as the chain length of the oil is increased. However, from the analysis of the surface tension data it is concluded that there are no major changes in the bending rigidities as the oil is changed. The data are analyzed within the model free energy densities [formula--see text] and [formula--see text]. We find that within experimental accuracy, the first of these models provides a quantitatively consistent description of the data. For the second model there is a larger discrepancy between observed and calculated values. PMID- 16711809 TI - Phase transition in two-dimensional dipolar fluids at low densities. AB - Monte Carlo computer simulations of a quasi two-dimensional (2D) dipolar fluid at low and intermediate densities indicate that the structure of the fluid is well described by an ideal mixture of self-assembling clusters. A detailed analysis of the topology of the clusters, of their internal energy and of their size (or mass) distributions is used to obtain approximations to their partition functions. Within the scope of these approximations, the results of this work suggest that the 2D dipolar fluid undergoes a phase transition from a dilute phase characterized by a number of disconnected clusters to a condensed phase characterized by a network or spanning (macroscopic) cluster that includes most of the particles in the system. PMID- 16711810 TI - Minimal model for chaotic shear banding in shear thickening fluids. AB - We present a minimal model for spatiotemporal oscillation and rheochaos in shear thickening complex fluids at zero Reynolds number. In the model, a tendency towards inhomogeneous flows in the form of shear bands combines with a slow structural dynamics, modeled by delayed stress relaxation. Using Fourier-space numerics, we study the nonequilibrium "phase diagram" of the fluid as a function of a steady mean (spatially averaged) stress, and of the relaxation time for structural relaxation. We find several distinct regions of periodic behavior (oscillating bands, traveling bands, and more complex oscillations) and also regions of spatiotemporal rheochaos. A low-dimensional truncation of the model retains the important physical features of the full model (including rheochaos) despite the suppression of sharply defined interfaces between shear bands. Our model maps onto the FitzHugh-Nagumo model for neural network dynamics, with an unusual form of long-range coupling. PMID- 16711811 TI - Size-dependent diffusion in an aging colloidal glass. AB - Colloidal dispersions of Laponite platelets are known to age slowly from viscous sols to colloidal glasses. We follow this aging process by monitoring the diffusion of probe particles embedded in the sample via dynamic light scattering. Our results show that the time-dependent diffusion of the probe particles scales with their size at early times and for the larger particles. This implies that within this regime the measurements can be used to investigate the generalized fluctuation-dissipation theorem for this out-of-equilibrium system, where the bath temperature is replaced with an effective temperature. Simultaneous dynamic rheological measurements reveal that this effective temperature increases as a function of aging time and frequency which suggests the existence of two measured time-scale regimes. In accord with recent work by Abou [Phys. Rev. Lett. 93, 160603 (2004)], our results suggest that at probed time scales longer than the characteristic relaxation time of the Laponite dispersion the system thermalizes with the bath, whereas at shorter time scales the system is out-of-equilibrium with an effective temperature greater than the bath temperature. PMID- 16711812 TI - Ultraslow dielectric relaxation process in supercooled polyhydric alcohols. AB - Complex permittivity was obtained on glycerol, xylitol, sorbitol and sorbitol xylitol mixtures in the supercooled liquid state in the frequency range between 10 microHz and 500 MHz at temperatures near and above the glass transition temperature. For all the materials, a dielectric relaxation process was observed in addition to the well-known structural alpha and Johari-Goldstein beta relaxation process [G. P. Johari and M. Goldstein, J. Chem. Phys. 53, 2372 (1970)]. The relaxation time for the new process is always larger than that for the alpha process. The relaxation time shows non-Arrhenius temperature dependence with correlation to the behavior of the alpha process and it depends on the molecular size systematically. The dielectric relaxation strength for the new process shows the effect of thermal history and decreases exponentially with time at a constant temperature. It can be considered that a nonequilibrium dynamics causes the new process. PMID- 16711813 TI - Gas adsorption and desorption in silica aerogels: a theoretical study of scattering properties. AB - We present a numerical study of the structural correlations associated with gas adsorption and desorption in silica aerogels in order to provide a theoretical interpretation of scattering experiments. Following our earlier work, we use a coarse-grained lattice-gas description and determine the nonequilibrium behavior of the adsorbed gas within a local mean-field analysis. We focus on the differences between the adsorption and desorption mechanisms and their signature in the fluid-fluid and gel-fluid structure factors as a function of temperature. At low temperature, but still in the regime where the isotherms are continuous, we find that the adsorbed fluid density, during both filling and draining, is correlated over distances that may be much larger than the gel correlation length. In particular, extended fractal correlations may occur during desorption, indicating the existence of a ramified cluster of vapor filled cavities. This also induces an important increase of the scattering intensity at small wave vectors. The similarity and differences with the scattering of fluids in other porous solids such as Vycor are discussed. PMID- 16711814 TI - Computing counterion densities at intermediate coupling. AB - By decomposing the Coulomb interaction into a long-distance component appropriate for mean-field theory, and a non-mean-field short distance component, we compute the counterion density near a charged surface for all values of the counterion coupling parameter. A modified strong-coupling expansion that is manifestly finite at all coupling strengths is used to treat the short-distance component. We find a nonperturbative correction related to the lateral counterion correlations that modifies the density at intermediate coupling. PMID- 16711815 TI - Confinement-induced instability of thin elastic film. AB - A confined incompressible elastic film does not deform uniformly when subjected to adhesive interfacial stresses but with undulations which have a characteristic wavelength scaling linearly with the thickness of the film. In the classical peel geometry, undulations appear along the contact line below a critical film thickness or below a critical curvature of the plate. Perturbation analysis of the stress equilibrium equations shows that for a critically confined film the total excess energy indeed attains a minimum for a finite amplitude of the perturbations which grow with further increase in the confinement. PMID- 16711816 TI - Vibration of submillimeter-size supported droplets. AB - We study the fundamental vibration mode of supported submillimeter-size droplets. Using an analogy with a simple oscillator we derive a semianalytical expression for the eigenfrequency and the scaling law of the energy dissipation within the droplet. The experimental results obtained for mercury drops deposited on glass are compared with the model. The agreement is satisfactory for the eigenfrequencies on the whole range of size we considered (from 0.04 to 0.9 mm). The scaling law for the dissipation is recovered for radii larger than 0.1 mm but fails for smaller droplets. We finally discuss possible applications related to the use of vibrations to effectively reduce the hysteresis of the wetting angle and therefore increase the mobility of the supported droplets. PMID- 16711817 TI - Wetting of rings on a nanopatterned surface: a lattice model study. AB - We perform mean-field density functional theory calculations on a lattice model to study the wetting of a solid substrate decorated with a ring pattern of nanoscale dimensions. We have found three different liquid morphologies on the substrate: a ring morphology where the liquid covers the pattern, a bulge morphology where a droplet is forming on one side of the ring, and a morphology where the liquid forms a cap spanning the nonwetting disk inside the pattern. We investigate the relative stability of these morphologies as a function of the ring size, wall-fluid interaction, and temperature. The results found are in very good agreement with experiments and calculations performed on similar systems at a micrometer length scale. The bulge morphology has also been observed in Monte Carlo simulations of the lattice model. Our results show that (i) morphologies of wetting patterns previously observed on a much larger (microm) scale can also form on a nm length scale, (ii) whether or not this happens depends crucially on the size of the wettable pattern, and (iii) the wettable ring may only be partially wet by the bulge morphology of the fluid. This morphology is a result of a spontaneously broken symmetry in the system. PMID- 16711818 TI - Effect of pressure on the phase behavior and structure of water confined between nanoscale hydrophobic and hydrophilic plates. AB - We perform systematic molecular dynamics simulations of water confined between two nanoscale plates at T = 300K. We investigate the effect of pressure (-0.15 GPa< or = P< or =0.2GPa) and plate separation (0.4 nm < or =d < or =1.6 nm) on the phase behavior of water when the plates are either hydrophobic or hydrophilic. When water is confined between hydrophobic plates, capillary evaporation occurs between the plates at low enough P. The threshold value of d at which this transition occurs decreases with P (e.g., 1.6 nm at P approximately equal to -0.05 GPa, 0.5 nm at P approximately equal to 0.1 GPa), until, at high P, no capillary evaporation occurs. For d approximately equal to 0.6 nm and P > or =0.1 GPa, the system crystallizes into a bilayer ice. A P-d phase diagram showing the vapor, liquid, and bilayer ice phases is proposed. When water is confined by hydrophilic (hydroxylated silica) plates, it remains in the liquid phase at all P and d studied. Interestingly, we observe for this case that even at the P at which bulk water cavitates, the confined water remains in the liquid state. We also study systematically the state of hydration at different P for both kinds of plates. For the range of conditions studied here, we find that in the presence of hydrophobic plates the effect of P is to enhance water structure and to push water molecules toward the plates. The average orientation of water molecules next to the hydrophobic plates does not change upon pressurization. In contrast, in the presence of hydrophilic plates, water structure is insensitive to P. Hence, our results suggest that upon pressurization, hydrophobic plates behave as "soft" surfaces (in the sense of accommodating pressure-dependent changes in water structure) while hydrophilic walls behave as "hard" surfaces. PMID- 16711819 TI - Coarse-graining a restricted solid-on-solid model. AB - A procedure suggested by Vvedensky for obtaining continuum equations as the coarse-grained limit of discrete models is applied to the restricted solid-on solid model with both adsorption and desorption. Using an expansion of the master equation, discrete Langevin equations are derived; these agree quantitatively with direct simulation of the model. From these, a continuum differential equation is derived, and the model is found to exhibit either Edwards-Wilkinson or Kardar-Parisi-Zhang exponents, as expected from symmetry arguments. The coefficients of the resulting continuum equation remain well-defined in the coarse-grained limit. PMID- 16711820 TI - Nonlocal hydrodynamic influence on the dynamic contact angle: slip models versus experiment. AB - Experiments reported by Blake [Phys. Fluids., 11, 1995 (1999)] suggest that the dynamic contact angle formed between the free surface of a liquid and a moving solid boundary at a fixed contact-line speed depends on the flow field and geometry near the moving contact line. We examine quantitatively whether or not it is possible to attribute this effect to the bending of the free surface due to hydrodynamic stresses acting upon it and hence interpret the results in terms of the so-called "apparent" contact angle. It is shown that this is not the case. Numerical analysis of the problem demonstrates that, at the spatial resolution reported in the experiments, the variations of the "apparent" contact angle (defined in two different ways) caused by variations in the flow field, at a fixed contact-line speed, are too small to account for the observed effect. The results clearly indicate that the actual (macroscopic) dynamic contact angle- i.e., the one used in fluid mechanics as a boundary condition for the equation determining the free surface shape--must be regarded as dependent not only on the contact-line speed but also on the flow field and geometry in the vicinity of the moving contact line. PMID- 16711821 TI - Quasidivergent nematic surface electroclinic effect. AB - A polyimide coated substrate is treated so that vertical liquid crystal alignment (theta=0) obtains over the temperature range T(NA)< T < T(a), where T(NA) is the nematic-smectic-A transition temperature. When the cell is filled with a chiral liquid crystal whose helical pitch is unwound (surface stabilized), application of an in-plane electric field for T(NA)< T < T(a) induces a nonzero polar tilt theta proportional to E of the liquid crystal director at the surface, where the tilted orientation propagates elastically into the bulk. On heating toward T(a), this surface electroclinic response becomes large, corresponding to the onset of a surface tilt transition at T(a) from theta=0 to nonzero theta. PMID- 16711822 TI - Polarization field effects at liquid-crystal-droplet-polymer interfaces. AB - The influence of confinement (droplet size) and liquid crystal orientational order (smectic-A and nematic) on the interfacial polarization field effects [Maxwell-Wagner-Sillars (MWS) effect] existing in liquid-crystal-droplets-polymer systems is investigated by broadband dielectric spectroscopy and a forward transmittance measurement technique. A relaxation process observed in the low frequency domain of the dielectric spectrum has been associated with a MWS effect for both micron-size and submicron-size droplets. Using electro-optical measurements and numerical simulations of the field inside droplets, it is shown that a depolarization field takes place in the same frequency range as that determined by dielectric spectroscopy. Differential scanning calorimetry measurements allowed to estimate the phase-separated liquid crystal [4,4' octylcyanobiphenyl (8CB)] fraction, which was found in the range of 55% for both micron-size and submicron-size droplets. X-ray diffraction experiments showed that smectic 8CB confined to micron-size cavities adopt bulklike properties, i.e., a partial bilayer structure, whereas in submicron-size droplets the layer spacing of the smectic phase is increased due to the strong bending deformations induced by the high curvature of the cavity walls. PMID- 16711823 TI - Temperature dependence of the Casimir-like force in free-standing smectic films. AB - The thermal Casimir-like force in free-standing liquid crystal films close to the smectic-A-nematic transition temperature is computed using a quadratic functional approach. In the framework of a microscopic mean-field model of free-standing smectic-A films, the temperature dependence of the order parameter profiles is computed and later used to estimate the elastic coupling variability in the vicinity of first- and second-order bulk smectic-A-nematic phase transitions. The strong nonuniformity of the coupling constant profiles promotes a significant increase of the fluctuation-induced force over three orders of magnitude, especially in thin films. This result reinforces the possible predominance of the thermal Casimir force as compared to the standard van der Waals interaction in thin smectic-A liquid crystal films. PMID- 16711824 TI - Surface-induced multiple reentrant transitions. AB - We have studied tilted surface layers in the bulk SmA temperature window of one liquid crystal using null transmission ellipsometry. Five distinct nonplanar surface structures were observed. We present analyses of the tilt and azimuthal profiles of the structures and examine the transitions between the structures. The transitions are identified as a double reentrant synclinic-anticlinic synclinic-anticlinic transition. Meanwhile, the transitions also display reentrant ferroelectric-antiferroelectric-ferroelectric behavior. PMID- 16711825 TI - Random anisotropy nematic model: nematic-non-nematic mixture. AB - The influence of a random-anisotropy- (RA-) type disorder on the phase separation of the nematogen-non-nematogen mixture is studied. A combination of the phenomenological Landau-de Gennes and Flory-Huggins theories is used. We assume that the non-nematogen component (i.e., impurity) enforces the RA disorder to the enclosing thermotropic liquid-crystal (LC) phase. The Imry-Ma argument is used according to which the lower-temperature phase exhibits a domain-type pattern. The disorder strength is measured in terms of the dimensionless parameter Lambda. We consider the case in which the LC molecules and impurities mix in the isotropic phase for Lambda=0. The impurities enforce a finite degree of orientational ordering even in the high-temperature paranematic phase. In the low temperature phase they give rise to a domain-type structure, resulting in the distorted nematic (speronematic) phase. We show that the onset of orientational ordering increases the phase separation tendency. The RA field, however, opposes this tendency. With increasing value of Lambda the difference between the paranematic and speronematic ordering decreases. Consequently the structure of the phase-separated pattern can be much more complex in comparison to the Lambda=0 case. PMID- 16711826 TI - Director configuration and self-organization of inclusions in two-dimensional smectic membranes. AB - The behavior of isolated inclusions (nematic droplets, smectic islands) and formation of chains and clusters from inclusions in oriented smectic membranes have been studied. Investigations of inclusions were performed in membranes in which the molecular ordering was oriented by an external magnetic field. At planar boundary conditions on the interface between the membrane and inclusions different configurations of the c-director field were observed: Coulombic, dipolar, quadrupolar, and mixed. The observed orientation of inclusions and their interactions and self-organization correlate with the predictions of the theory based on the electromagnetic analogy. Chaining and formation of superstructures differ in oriented and nonoriented membranes. PMID- 16711827 TI - Stress-guided self-assembly in Dutcher films. AB - Dutcher films consist of a layer of liquid sandwiched between two solid capping layers and can spontaneously self-assemble to form corrugated surfaces. The interplay between the attractive van der Waals forces across the film, and the elastic forces due to the deformation of the capping layers, produces well defined periodic undulations. We show how computer simulations can capture both the formation of undulations in Dutcher films and the correct periodicity. Furthermore, we simulate Dutcher films which are either compressed or stretched, resulting in the promotion or suppression of undulation growth. In this manner, applied deformations can be shown to guide the self-assembly process in Dutcher films and result in the formation of highly oriented surface corrugations over large distances. PMID- 16711828 TI - Substrate adhesion of a nongrafted flexible polymer in a cavity. AB - In a contact-density chain-growth study we investigate the solubility-temperature pseudophase diagram of a lattice polymer in a cavity with an attractive surface. In addition to the main phases of adsorbed and desorbed conformations we find numerous subphases of collapsed and expanded structures. PMID- 16711829 TI - Micelle formation in statistical copolymers. AB - Formation of microdomain structures in concentrated systems of irregular (stochastic) copolymers with weak composition asymmetry (epsilon>>1) is considered theoretically. The study is focused on the weak segregation regime for infinitely long copolymer chains near the disorder-to-order transition. It is shown that the transition occurs below the spinodal (at chi(0)< chi*) and that it results in the formation of micelles rather than in a superposition pattern of a few harmonic composition waves. The micelle size is inversely proportional to Delta(chi=chi)-(chi0). For small Delta(chi) the micelles are like large nearly uniform droplets with "reflected" composition, -epsilon. As Delta(chi) increases the micelle composition profile develops oscillations. A first-order transition from spherical-wave micelles to micelles with internal bcc structure is predicted at chi/chi*-1 proportional to epsilon(2). Interaction of micelles is also considered. It is shown that micelles always tend to form a fcc superlattice. The micellization in the weak segregation regime is controlled by the so-called nonlocal free energy. The classical fourth order (in composition parameter A) approximation for this energy is significantly generalized in the regime of low volume fraction of micelles. It is shown that the nonlocal energy strongly (exponentially) increases with A near and above a certain critical value A approximately equal to A*. PMID- 16711830 TI - Improving the speed of the genetic toggle switch without sacrificing its dynamic stability. AB - Determinants of the switching speed of the genetic toggle switch remain unknown. Analysis shows that the decay rate of proteins predominantly sets the speed limit, but its modification introduces a trade-off between increased speed and decreased bistability. Incorporating protein-modifying enzymes into the switch gives extra degrees of freedom to address this trade-off. The condition for bistability when such enzymes are incorporated is derived. Under this condition, speed increases with the maximal rate of gene expression. PMID- 16711831 TI - Dynamical response of the Hodgkin-Huxley model in the high-input regime. AB - The response of the Hodgkin-Huxley neuronal model subjected to stochastic uncorrelated spike trains originating from a large number of inhibitory and excitatory post-synaptic potentials is analyzed in detail. The model is examined in its three fundamental dynamical regimes: silence, bistability, and repetitive firing. Its response is characterized in terms of statistical indicators (interspike-interval distributions and their first moments) as well as of dynamical indicators (autocorrelation functions and conditional entropies). In the silent regime, the coexistence of two different coherence resonances is revealed: one occurs at quite low noise and is related to the stimulation of subthreshold oscillations around the rest state; the second one (at intermediate noise variance) is associated with the regularization of the sequence of spikes emitted by the neuron. Bistability in the low noise limit can be interpreted in terms of jumping processes across barriers activated by stochastic fluctuations. In the repetitive firing regime a maximization of incoherence is observed at finite noise variance. Finally, the mechanisms responsible for the different features appearing in the interspike-interval distributions (like multimodality and exponential tails) are clearly identified in the various regimes. PMID- 16711832 TI - Scaling relations in food webs. AB - In the last three decades, researchers have tried to identify universal patterns in the structure of food webs. It was recently proposed that the exponent eta characterizing the efficiency of the transport of energy in large and small food webs might have a universal value (eta = 1.13). In this work we establish lower and upper bounds for this exponent in a general spanning tree with a fixed number of trophic species and levels. When the number of species is large, the lower and upper bounds are equal to 1, implying that the result eta = 1.13 is due to finite size effects and that the value of this exponent depends on the size of the web. We also evaluate analytically and numerically the exponent eta for hierarchical and random networks. In all cases the exponent eta depends on the number of trophic species K, and when K is large we have that eta-->1. Moreover, this result holds for any fixed number M of trophic levels. PMID- 16711833 TI - Patchy propagators, brain dynamics, and the generation of spatially structured gamma oscillations. AB - Propagator theory of brain dynamics is generalized to incorporate a new class of patchy propagators that enable treatment of approximately periodic structures such as are seen in the visual cortex. Complex response fields are also incorporated to allow for features such as orientation preference and wave-number selectivity. The results are applied to the corticothalamic system associated with the primary visual cortex. It is found that this system can generate gamma ( > or = 30 Hz) oscillations during stimulation, whose properties are consistent with experimental findings on gamma frequency and bandwidth, and existence of fine-scale spatial structure. It is found that a potential resonance is associated with each reciprocal lattice vector corresponding to periodic modulations of the propagators. It is found that the lowest resonances are the most likely to give rise to noticeable spectral peaks and increases of correlation amplitude, length, and time, and that these aspects are prominent only if the system is close to marginal stability, in accord with previous measurements and discussions of cortical stability. These features also enable gamma resonances to be stimulus-evoked, with substantial resonance sharpening for relatively small changes in mean neural firing rate. The results also imply dependence of gamma frequency on stimulus features. PMID- 16711834 TI - Spectral filtering of visible light by the cuticle of metallic woodboring beetles and microfabrication of a matching bioinspired material. AB - Samples of the cuticle taken from the body of Buprestidae Chrysochroa vittata have been studied by scanning electron microscopy and optical reflectance measurements, related to numerical simulations. The cause of the metallic coloration of the body of these insects is determined to be the structure of the hard carapace constructed as a stack of thin chitin layers separated by very thin irregular air gaps. In particular the change of color as a function of the observation angle is elucidated in terms of an infinite photonic-crystal model, confirmed by finite multilayer calculations. These mechanisms are used to develop an artificial bioinspired multilayer system which reproduces the visual effects provided by the insect surface. PMID- 16711835 TI - Dynamics of moment neuronal networks. AB - A theoretical framework is developed for moment neuronal networks (MNNs). Within this framework, the behavior of the system of spiking neurons is specified in terms of the first- and second-order statistics of their interspike intervals, i.e., the mean, the variance, and the cross correlations of spike activity. Since neurons emit and receive spike trains which can be described by renewal--but generally non-Poisson--processes, we first derive a suitable diffusion-type approximation of such processes. Two approximation schemes are introduced: the usual approximation scheme (UAS) and the Ornstein-Uhlenbeck scheme. It is found that both schemes approximate well the input-output characteristics of spiking models such as the IF and the Hodgkin-Huxley models. The MNN framework is then developed according to the UAS scheme, and its predictions are tested on a few examples. PMID- 16711836 TI - Large-scale dimension densities for heart rate variability analysis. AB - In this work, we reanalyze the heart rate variability (HRV) data from the 2002 Computers in Cardiology (CiC) Challenge using the concept of large-scale dimension densities and additionally apply this technique to data of healthy persons and of patients with cardiac diseases. The large-scale dimension density (LASDID) is estimated from the time series using a normalized Grassberger Procaccia algorithm, which leads to a suitable correction of systematic errors produced by boundary effects in the rather large scales of a system. This way, it is possible to analyze rather short, nonstationary, and unfiltered data, such as HRV. Moreover, this method allows us to analyze short parts of the data and to look for differences between day and night. The circadian changes in the dimension density enable us to distinguish almost completely between real data and computer-generated data from the CiC 2002 challenge using only one parameter. In the second part we analyzed the data of 15 patients with atrial fibrillation (AF), 15 patients with congestive heart failure (CHF), 15 elderly healthy subjects (EH), as well as 18 young and healthy persons (YH). With our method we are able to separate completely the AF (rho (mu/ls) = 0.97 +/- 0.02) group from the others and, especially during daytime, the CHF patients show significant differences from the young and elderly healthy volunteers (CHF, 0.65 +/- 0.13; EH, 0.54 +/- 0.05; YH, 0.57 +/- 0.05; p < 0.05 for both comparisons). Moreover, for the CHF patients we find no circadian changes in rho (mu/ls) (day, 0.65 +/- 0.13; night, 0.66 +/- 0.12; n.s.) in contrast to healthy controls (day, 0.54 +/- 0.05; night, 0.61 +/- 0.05; p=0.002). Correlation analysis showed no statistical significant relation between standard HRV and circadian LASDID, demonstrating a possibly independent application of our method for clinical risk stratification. PMID- 16711837 TI - Elasticity of peptide omega bonds. AB - We calculated the changes of the free energy profile of the peptidyl-prolyl torsional angle of the dipeptide valine-proline under pulling forces by simulations. Using a dynamic model built on the equilibrium properties of this system and previously studied dynamic properties of cis-trans isomerization of other dipeptides, we calculated the dynamic viscoelasticity of this degree of freedom. The results show significant differences between how thermal and mechanical forces alter the equilibrium and the dynamics of the isomerization transition. The former does not change the barrier heights but changes the prefactor of the kinetics owing to temperature effects, while the latter changes minima and thus the population. The force that is required to "excite" this degree of freedom is small. Compared to other systems, we found that this degree of freedom becomes already quite rigid at several hertz, which is a much lower value due to the high barrier of the cis-trans isomerization. We also found that the tensile elastic modulus of densely packed omega bonds is at the order of GPa, which is comparable to that of polymer materials. These results give mechanical properties of polyproline elasticity of a local nature and provide guidance for future experimental designs. PMID- 16711838 TI - Powering a burnt bridges Brownian ratchet: a model for an extracellular motor driven by proteolysis of collagen. AB - Biased diffusion of collagenase on collagen fibrils may represent the first observed adenosine triphosphate-independent extracellular molecular motor. The magnitude of force generated by the enzyme remains unclear. We propose a propulsion mechanism based on a burnt bridges Brownian ratchet model with a varying degree of coupling of the free energy from collagen proteolysis to the enzyme motion. When constrained by experimental observations, our model predicts 0.1 pN stall force for individual collagenase molecules. A dimer, surprisingly, can generate a force in the range of 5 pN, suggesting that the motor can be of biological significance. PMID- 16711839 TI - Observation of all the components of elastic constants using tetragonal hen egg white lysozyme crystals dehydrated at 42% relative humidity. AB - Success in measuring transverse sound velocity allowed us to determine, for the first time, all six elastic constants of a protein crystal. An ultrasonic pulse echo method was used to perform sound velocity measurements on tetragonal hen egg white (HEW) lysozyme crystals that were partially dehydrated at 42% relative humidity. The measurements were performed using the (110), (101), and (001) crystallographic faces. Thus, all six elastic constants of the dehydrated tetragonal HEW lysozyme crystals were determined: C11=C22=12.44 GPa, C12=7.03 GPa, C13=C23=8.36 GPa, C33=12.79 GPa, C44=C55=2.97 GPa, and C66=2.63 GPa. In addition, for the hydrated crystals, the longitudinal sound velocities along the [110] direction and the direction normal to the (101) face were measured. From these results, all the components of elastic constants in the hydrated crystals were extrapolated. PMID- 16711840 TI - Learning the structure of correlated synaptic subgroups using stable and competitive spike-timing-dependent plasticity. AB - Synaptic plasticity must be both competitive and stable if ongoing learning of the structure of neural inputs is to occur. In this paper, a wide class of spike timing-dependent plasticity (STDP) models is identified that have both of these desirable properties in the case in which the input consists of subgroups of synapses that are correlated within the subgroup through the occurrence of simultaneous input spikes. The process of synaptic structure formation is studied, illustrating one particular class of these models. When the learning rate is small, multiple alternative synaptic structures are possible given the same inputs, with the outcome depending on the initial weight configuration. For large learning rates, the synaptic structure does not stabilize, resulting in neurons without consistent response properties. For learning rates in between, a unique and stable synaptic structure typically forms. When this synaptic structure exhibits a bimodal distribution, the neuron will respond selectively to one or more of the subgroups. The robustness with which this selectivity develops during learning is largely determined by the ratio of the subgroup correlation strength to the number of subgroups. The fraction of potentiated subgroups is primarily determined by the balance between potentiation and depression. PMID- 16711841 TI - Intercellular waves propagation in an array of cells coupled through paracrine signaling: a computer simulation study. AB - A linear chain of cells is considered in which calcium (Ca2+) fluctuations within a cell are described by a simple minimal model. Cells are coupled together by bidirectional paracrine signaling via calcium oscillations. Two typical zones of propagation are observed: a transition zone and a regular zone. The transition zone exhibits the same phenomena that can be observed in single cells, pairs or triplets of cells. Within the regular zone, simple periodic oscillations of calcium propagate and the Ca2+ signal is similar from one cell to another (same amplitude and same frequency). But, the signals are separated by a slight phase shift characterizing the propagation of Ca2+ waves due to the type of coupling used. We also consider the colonization of the lattice by the abnormal oscillations of sick cells. PMID- 16711842 TI - Quasispecies theory for multiple-peak fitness landscapes. AB - We use a path integral representation to solve the Eigen and Crow-Kimura molecular evolution models for the case of multiple fitness peaks with arbitrary fitness and degradation functions. In the general case, we find that the solution to these molecular evolution models can be written as the optimum of a fitness function, with constraints enforced by Lagrange multipliers and with a term accounting for the entropy of the spreading population in sequence space. The results for the Eigen model are applied to consider virus or cancer proliferation under the control of drugs or the immune system. PMID- 16711843 TI - Blocking of an ion channel by a highly charged drug: modeling the effects of applied voltage, electrolyte concentration, and drug concentration. AB - We present a simple physical model to estimate the blocked pore probability of an ion channel that can be blocked by a highly charged drug in solution. The model is inspired by recent experimental work on the blocking of the PA(63) channel, involved in the anthrax toxin infection, by a highly charged drug [Karginov PNAS 102, 15075 (2005)]. The drug binding to the pore is highly specific but the strong dependence of blocking on the applied voltage and electrolyte concentration suggests that long range electrostatic interactions are important. Since basic electrostatic concepts rather than detailed molecular models are considered, the microscopic details of the channel blocking are ignored, although the model captures most of the qualitative characteristics of the problem. PMID- 16711844 TI - Coupling between vesicle shape and lateral distribution of mobile membrane inclusions. AB - Membrane inclusions such as membrane-embedded peptides or proteins exhibit a curvature-dependent interaction with the surrounding lipid matrix due to the mismatch between their intrinsic curvature and the local membrane curvature. This interaction causes an inhomogeneous lateral distribution of the inclusions and a corresponding adjustment of the vesicle shape. We have studied theoretically the axisymmetric equilibrium shapes of lipid vesicles with mobile inclusions, taking into account that the membrane free energy includes the elastic energy of the lipid bilayer and a contribution due to an inclusion-membrane interaction. Equations describing the shape are derived by minimizing the total free energy at fixed membrane area, enclosed volume, and number of inclusions and are then solved numerically. It is shown that vesicle shape may assume a symmetry that differs from that of the vesicle with no inclusions. If the inclusion-membrane interaction exceeds a certain value, there is no axisymmetric solution of the equations with a continuous and derivable lateral density of inclusions over the whole area of the vesicle. When approaching the critical vesicle shape, the shapes obtained differ qualitatively from those described by the area difference elasticity model of the elastic properties of lipid membranes. In general, vesicle shapes adjust to the presence of inclusions by increasing regions with favorable curvature and decreasing regions of unfavorable curvature in a way such that the lateral distribution of inclusions becomes inhomogeneous. PMID- 16711845 TI - Optical properties of the iridescent organ of the comb-jellyfish Beroe cucumis (Ctenophora). AB - Using transmission electron microscopy, analytical modeling, and detailed numerical simulations, the iridescence observed from the comb rows of the ctenophore Beroe cucumis was investigated. It is shown that the changing coloration which accompanies the beating of comb rows as the animal swims can be explained by the weakly-contrasted structure of the refractive index induced by the very coherent packing of locomotory cilia. The colors arising from the narrow band-gap reflection are shown to be highly saturated and, as a function of the incidence angle, cover a wide range of the visible and ultraviolet spectrum. The high transparency of the structure at the maximal bioluminescence wavelength is also explained. PMID- 16711846 TI - Role of repulsive factors in vascularization dynamics. AB - Capillary networks are essential in vertebrates to supply tissues with nutrients. Experiments of in vitro capillary formation show that endothelial cells randomly spread on a gel matrix autonomously organize to form vascular networks with a characteristic length independent of the initial cell density. A mathematical model based on free cell migration and on cell cross-talk mediated by soluble chemical factors has been recently proposed and explains the main dynamical and geometrical properties of the networks. We extend this model introducing the action of repulsive factors and we show that their activity results in a larger degree of reorganization of cellular matter and in more robust control over the size of the growing vascular network. PMID- 16711847 TI - Diffusion in curved fluid membranes. AB - We analyze theoretically the effects of curvature on the diffusion in a fluid membrane, within the Saffman-Delbruck hydrodynamic model. We calculate the effect of curvature on the intrinsic fluidity of a membrane through changes in its thickness, for both static or fluctuating curvature. We treat both thermal curvature fluctuations, and fluctuations due to active processes. Such curvature fluctuations increase the average membrane thickness and diminish the projected area, thereby decreasing the diffusion coefficient. This calculation allows us to predict the effect of shear flow on the membrane diffusion, and to compare to observations on living cells. PMID- 16711848 TI - Intramolecular dynamics of linear macromolecules by fluorescence correlation spectroscopy. AB - A theoretical description of the dynamics of DNA molecules and actin filaments in solution as measured experimentally by fluorescence correlation spectroscopy is provided and compared to recent experimental results. Particular attention is paid to the contribution of the intramolecular dynamics to the fluorescence correlation function. Using a semiflexible chain model, a theoretical expression is presented for the fluorescence correlation spectroscopy correlation function. The dependence of this function on various model parameters, such as chain length, persistence length, and fluorescence label density, is discussed. Our investigations show that the intramolecular dynamics provides a significant contribution or even dominates the correlation function as soon as the longest intramolecular relaxation time significantly exceeds the shortest experimentally accessible time. Correspondingly, the shape of the correlation function changes considerably. Approximate analytical expressions are provided, which are in qualitative agreement with the exact theoretical solutions as well as experimental results, for both DNA and actin filaments. Our approach is in agreement with the predictions of the Zimm model, in the limit of very flexible polymers, as well as the predictions of semiflexible polymer models with respect to the intramolecular dynamics in solution. PMID- 16711849 TI - Classification of epilepsy types through global network analysis of scalp electroencephalograms. AB - Epilepsy is a dynamic disease in which self-organization and emergent structures occur dynamically at multiple levels of neuronal integration. Therefore, the transient relationship within multichannel electroencephalograms (EEGs) is crucial for understanding epileptic processes. In this paper, we show that the global relationship within multichannel EEGs provides us with more useful information in classifying two different epilepsy types than pairwise relationships such as cross correlation. To demonstrate this, we determine the global network structure within channels of the scalp EEG based on the minimum spanning tree method. The topological dissimilarity of the network structures from different types of temporal lobe epilepsy is described in the form of the divergence rate and is computed for 11 patients with left (LTLE) and right temporal lobe epilepsy (RTLE). We find that patients with LTLE and RTLE exhibit different large scale network structures, which emerge at the epoch immediately before the seizure onset, not in the preceding epochs. Our results suggest that patients with the two different epilepsy types display distinct large scale dynamical networks with characteristic epileptic network structures. PMID- 16711850 TI - Monte Carlo simulations of polyelectrolytes inside viral capsids. AB - Structural features of polyelectrolytes as single-stranded RNA or double-stranded DNA confined inside viral capsids and the thermodynamics of the encapsidation of the polyelectrolyte into the viral capsid have been examined for various polyelectrolyte lengths by using a coarse-grained model solved by Monte Carlo simulations. The capsid was modeled as a spherical shell with embedded charges and the genome as a linear jointed chain of oppositely charged beads, and their sizes corresponded to those of a scaled-down T=3 virus. Counterions were explicitly included, but no salt was added. The encapisdated chain was found to be predominantly located at the inner capsid surface, in a disordered manner for flexible chains and in a spool-like structure for stiff chains. The distribution of the small ions was strongly dependent on the polyelectrolyte-capsid charge ratio. The encapsidation enthalpy was negative and its magnitude decreased with increasing polyelectrolyte length, whereas the encapsidation entropy displayed a maximum when the capsid and polyelectrolyte had equal absolute charge. The encapsidation process remained thermodynamically favorable for genome charges ca. 3.5 times the capsid charge. The chain stiffness had only a relatively weak effect on the thermodynamics of the encapsidation. PMID- 16711851 TI - Stochastic model of coliphage lambda regulatory network. AB - The dynamic properties of the regulatory network governing the choice between lytic and lysogenic growths of coliphage lambda is studied using a Markov chain stochastic model. Our computer simulation confirms the finding by Li et al. [Proc. Natl. Acad. Sci. USA 101, 4781 (2004)] on the dynamics of budding yeast: that the biological stationary states are global attractors; the biological pathways of lytic and lysogenic growths are attracting trajectories; and the network functions are robustly designed against structural perturbations. In addition, our model shows the stochastic switch from lysogen to lytic growth, which has been observed in experiments. A definition of pseudoenergy is introduced in the network dynamics to reveal a transitionlike behavior in the system. PMID- 16711852 TI - Enhancement of internal-noise coherence resonance by modulation of external noise in a circadian oscillator. AB - A circadian oscillator driven by external noise and internal noise has been studied by use of the chemical Langevin equation method. When the system is near a Hopf bifurcation and driven by internal noise only, it is found that the coherence resonance phenomenon can be induced by the internal noise. When the system is simultaneously driven by internal and external noise, it is found that external-noise coherence resonance can be suppressed by internal noise, while internal-noise coherence resonance can be enhanced by modulation of the external noise intensity in a certain range of noise intensity. Another interesting result is that the external noise can regulate the optimal system size when the internal noise coherence resonance occurs. PMID- 16711853 TI - Bifurcation analysis of mode-locking structure in a Hodgkin-Huxley neuron under sinusoidal current. AB - Nervous systems under periodic stimuli display rich dynamical states including mode-locking and chaotic responses, which have been a subject of intense studies in neurodynamics. The bifurcation structure of the Hodgkin-Huxley neuron under sinusoidal stimulus is studied in detail. The mechanisms of the firing onset and rich firing dynamics are studied with the help of the codimension-2 bifurcations, which play the role of the organizing center for myriads of saddle-node, period doubling, and inverse-flip bifurcations forming the boundaries of the complex mode-locking structure. This study provides a useful insight into the organization of similar bifurcation structures in excitable systems such as neurons under periodic forcing. PMID- 16711854 TI - Graph theoretic properties of networks formed by the Delaunay tessellation of protein structures. AB - The Delaunay tessellation of several sets of real and simplified model protein structures has been used to explore graph theoretic properties of residue contact networks. The system of contacts defined by residues joined by edges in the Delaunay simplices can be thought of as a graph or network and analyzed using techniques from elementary graph theory and the theory of complex networks. Such analysis indicates that protein contact networks have small world character, but technically are not small world networks. This approach also indicates that networks formed by native structures and by most misfolded decoys can be differentiated by their respective graph properties. The characteristic features of residue contact networks can be used for the detection of structural elements in proteins, such as the ubiquitous closed loops consisting of 22-32 consecutive residues, where terminal residues are Delaunay neighbors. PMID- 16711855 TI - Identification of key residues in proteins by using their physical characters. AB - Key residues in proteins are important to their stability, folding, and functions. They usually are highly conserved and can be identified by sequence or structure alignments. However, these methods can only determine the locations of key residues in sequences and structures and give less information about their physical characters. In this paper, we try to identify key residues by analyzing their inter-residue interactions. The model we study is the protein domain from transducin. We show that the usual Gaussian network analysis and distance-based contact analysis have difficulty identifying the key residues in this protein, but the contact energies can do it well. We find that most key residues can be located by the lowest contact energies. This enables us to predict and analyze the key residues in other proteins. Our results suggest that contact energy analysis may provide an alternative approach to investigating the folding and stability of proteins. PMID- 16711857 TI - Bifurcation control of a seizing human cortex. AB - We consider as a mathematical model of human cortical electrical activity a system of fourteen ordinary differential equations. With appropriate parameters, the model produces activity characteristic of a seizure. To prevent such seizures, we incorporate feedback controllers into the model dynamics. We show that three controllers--a linear feedback controller, a differential controller, and a filter controller--can be used to eliminate seizing activity in the model system. We show how bifurcations induced by the linear controller alter those present in the original dynamics. PMID- 16711856 TI - Monte Carlo simulation of chromatin stretching. AB - We present Monte Carlo (MC) simulations of the stretching of a single chromatin fiber. The model approximates the DNA by a flexible polymer chain with Debye Huckel electrostatics and uses a two-angle zigzag model for the geometry of the linker DNA connecting the nucleosomes. The latter are represented by flat disks interacting via an attractive Gay-Berne potential. Our results show that the stiffness of the chromatin fiber strongly depends on the linker DNA length. Furthermore, changing the twisting angle between nucleosomes from 90 degrees to 130 degrees increases the stiffness significantly. An increase in the opening angle from 22 degrees to 34 degrees leads to softer fibers for small linker lengths. We observe that fibers containing a linker histone at each nucleosome are stiffer compared to those without the linker histone. The simulated persistence lengths and elastic moduli agree with experimental data. Finally, we show that the chromatin fiber does not behave as an isotropic elastic rod, but its rigidity depends on the direction of deformation: Chromatin is much more resistant to stretching than to bending. PMID- 16711858 TI - Stimulus-response curves of a neuronal model for noisy subthreshold oscillations and related spike generation. AB - We investigate the stimulus-dependent tuning properties of a noisy ionic conductance model for intrinsic subthreshold oscillations in membrane potential and associated spike generation. Upon depolarization by an applied current, the model exhibits subthreshold oscillatory activity with an occasional spike generation when oscillations reach the spike threshold. We consider how the amount of applied current, the noise intensity, variation of maximum conductance values, and scaling to different temperature ranges alter the responses of the model with respect to voltage traces, interspike intervals and their statistics, and the mean spike frequency curves. We demonstrate that subthreshold oscillatory neurons in the presence of noise can sensitively and also selectively be tuned by the stimulus-dependent variation of model parameters. PMID- 16711860 TI - Noise-induced oscillatory behavior in field-dependent relaxational dynamics. AB - By considering inertial effects in a field-dependent relaxational model, we show that noise may induce collective oscillatory dynamics. In agreement with the recently introduced idea of noise-induced multistability, we show that there is a region in parameter space where such behavior depends on the initial condition. Moreover, when the coupling term leads to pattern formation by means of a morphological instability a la Swift-Hohenberg, [J. Buceta, M. Ibanes, J. M. Sancho, and K. Lindenberg, Phys. Rev. E 67, 021113 (2003) and K. Wood, J. Buceta, and K. Lindenberg, Phys. Rev. E 73, 022101 (2006)] our numerical simulations reveal that spatio-temporal oscillatory structures develop. PMID- 16711861 TI - Regulating noise-induced spiking using feedback. AB - We report successful manipulation of the noise provoked spiking behavior using delayed feedback control. Experiments were performed in a three electrode electrochemical cell under potentiostatic conditions. The uncontrolled system exhibited noise invoked oscillations whose regularity was quantified using normalized variance (NV). Superimposing delayed feedback, for appropriate values of delay (t), an enhancement in the regularity of the spike sequence was attained. Numerical simulations corroborated experimental observations. PMID- 16711859 TI - Voltage-dependent capacitance of human embryonic kidney cells. AB - We determine membrane capacitance, C as a function of dc voltage for the human embryonic kidney (HEK) cell. C was calculated from the admittance, Y, obtained during a voltage ramp when the HEK cell was held in whole-cell patch-clamp configuration. Y was determined at frequencies of 390.625 and from the measured current, i obtained with a dual-sinusoidal stimulus. We find that the fractional increase in the capacitance, C is small ( < 1%) and grows with the square of the voltage, Psi. C can be described by: C=C(0)(1+alpha(Psi+psi(s))2)[where C(0): Capacitance at 0 volts, psi(s): Difference in surface potential between cytoplasmic and extracellular leaflets and alpha: Proportionality constant]. We find that alpha and psi(s) are 0.120 (+/- 0.01) V(-2) and -0.073 (+/-0.017 V in solutions that contain ion channel blockers and 0.108 (+/- 0.29) V(-2) and -0.023 (+/- 0.009) V when 10 mM sodium salicylate was added to the extracellular solution. This suggests that salicylate does not affect the rate at which C grows with Psi, but reduces the charge asymmetry of the membrane. We also observe an additional linear differential capacitance of about (-46 fFV(-1)) in about 60% of the cells, this additional component acts simultaneously with the quadratic component and was not observed when salicylate was added to the solution. We suggest that the voltage dependent capacitance originates from electromechanical coupling either by electrostriction and/or Maxwell stress effects and estimate that a small electromechanical force (approximately equal to 1 pN) acts at physiological potentials. These results are relevant to understand the electromechanical coupling in outer hair cells (OHCs) of the mammalian cochlea, where an asymmetric bell-shaped C versus Psi relationship is observed upon application of a similar field. Prestin, a membrane protein expressed in OHCs is required to observe this function. When we compare the total charge contributions from HEK cell membrane (7 x 10(4) electrons, 10 pF cell) with that determined for prestin transfected cells (up to 5 x 10(6) electrons) we conclude that the charge contributions from the collective motion of membrane proteins and lipids in the field is dwarfed relative to that when prestin is present. We suggest that the capacitance-voltage relationships should be similar to that observed for HEK cells for OHCs that do not express prestin in their membranes. PMID- 16711862 TI - Topological transitions in two-dimensional lattice spin models. AB - A family of classical statistical-mechanical spin models, by now extensively studied in the literature, involves two-component unit vectors, associated with a two-dimensional lattice, with pair potentials restricted to nearest neighbors and possessing 0(2) symmetry--i.e., defined by some function of the scalar product between the two interacting spins; these studies often show the existence of a topological phase transition. We show here that, for a wide class of interaction models of the above type, available mathematical results entail the existence of a topological (Berezinskii-Kosterlitz-Thouless-like) transition, as well as a rigorous lower bound on the transition temperature. PMID- 16711863 TI - Mass fractal dimension of the ribosome and implication of its dynamic characteristics. AB - Self-similar properties of the ribosome in terms of the mass fractal dimension are investigated. We find that both the 30S subunit and the 16S rRNA have fractal dimensions of 2.58 and 2.82, respectively; while the 50S subunit as well as the 23S rRNA has the mass fractal dimension close to 3, implying a compact three dimensional macromolecule. This finding supports the dynamic and active role of the 30S subunit in the protein synthesis, in contrast to the pass role of the 50S subunit. PMID- 16711864 TI - Heat fluctuations in Brownian transducers. AB - The heat fluctuation probability distribution function in Brownian transducers operating between two heat reservoirs is studied. We find, both analytically and numerically, that the recently proposed fluctuation theorem for heat exchange [C. Jarzynski and D. K. Wojcik, Phys. Rev. Lett. 92, 230602 (2004)] has to be applied carefully when the coupling mechanism between both baths is considered. We also conjecture how to extend such a relation when an external work is present. PMID- 16711865 TI - Predictions of bond percolation thresholds for the kagome and Archimedean (3, 12(2)) lattices. AB - Here we show how the recent exact determination of the bond percolation threshold for the martini lattice can be used to provide approximations to the unsolved kagome and (3, 12(2)) lattices. We present two different methods: one which provides an approximation to the inhomogeneous kagome and bond problems, and the other which gives estimates of for the homogeneous kagome (0.524 408 8...) and (3, 12(2)) (0.740 421 2...) problems that, respectively, agree with numerical results to five and six significant figures. PMID- 16711866 TI - Synchronization and symmetry breaking in mutually coupled fiber lasers. AB - We experimentally study the synchronization and the emergence of leader-follower dynamics in two time-delayed mutually coupled fiber ring lasers. We utilize spatiotemporal representations of time series to establish the roles of leader and follower in the synchronized dynamics. PMID- 16711867 TI - Oscillating droplets by decomposition on the spherical harmonics basis. AB - In the framework of Rayleigh's description, we have investigated the eigenfrequencies of the capillary waves of a nonwetting droplet under forced oscillations (pointlike force). The theoretical model using the spherical harmonics Y(l,m)(theta, phi) as a part of the solution of the Laplace equation, is in good agreement with the experimental results. This model can be generalized for all kinds of excitations with a sitting or a levitating droplet due to the decomposition of the excitation on the spherical harmonics basis. From this study, a different theoretical way of interpreting droplet bouncing is presented motivating a wide range of industrial applications. PMID- 16711868 TI - Surface roughness and hydrodynamic boundary conditions. AB - We report results of investigations of a high-speed drainage of thin aqueous films squeezed between randomly nanorough surfaces. A significant decrease in the hydrodynamic resistance force as compared with that predicted by Taylor's equation is observed. However, this reduction in force does not represent the slippage. The measured force is exactly the same as that between equivalent smooth surfaces obeying no-slip boundary conditions, but located at the intermediate position between peaks and valleys of asperities. The shift in hydrodynamic thickness is shown to be independent of the separation and/or shear rate. Our results disagree with previous literature data reporting very large and shear-dependent boundary slip for similar systems. PMID- 16711869 TI - Fluctuations of local density of states and C0 speckle correlations are equal. AB - We establish a conceptual relation between the fluctuations of the local density of states (LDOS) and the intensity correlations in speckle patterns resulting from the multiple scattering of waves in random media. We show that among the known types of speckle correlations (C1, C2, C3, and C0) only contributes to LDOS fluctuations in the infinite medium. We propose to exploit the equivalence of LDOS fluctuations and the C0 intensity correlation as a "selection rule" for scattering processes contributing to C0. PMID- 16711870 TI - Observation of fast light in Mie scattering processes. AB - We observed -4.9 nsec negative group delay in the optical wave-packet propagation through a fiber taper coupled with a single Mie sphere. The observed fast light under the Mie scattering process agrees excellently with the anomalous dispersion calculated on the basis of a directional coupling theory on the undercoupling condition, which also predicts a positive group delay on the overcoupling condition. PMID- 16711871 TI - Nontopological Raman-Kerr self-induced transparency solitons in photonic crystal fibers. AB - Using analytical and numerical methods we demonstrate that the pulse propagation equations accounting for the quantum description of the Raman transition and for the optical Kerr nonlinearity have a multiparameter family of nontopological solitary wave solutions. We study properties of these solitons and report the transition from stable to unstable regimes of propagation. We also discuss the feasibility of observation of these structures in gas filled hollow-core photonic crystal fibers. PMID- 16711872 TI - Neighborhood properties of complex networks. AB - A concept of neighborhood in complex networks is addressed based on the criterion of the minimal number of steps to reach other vertices. This amounts to, starting from a given network R1, generating a family of networks Rl, l = 2, 3,... such that, the vertices that are l steps apart in the original R1, are only 1 step apart in Rl. The higher order networks are generated using Boolean operations among the adjacency matrices Ml that represent Rl. The families originated by the well known linear and the Erdos-Renyi networks are found to be invariant, in the sense that the spectra of Ml are the same, up to finite size effects. A further family originated from small world network is identified. PMID- 16711873 TI - Distribution of edge load in scale-free trees. AB - Node betweenness has been studied recently by a number of authors, but until now less attention has been paid to edge betweenness. In this paper, we present an exact analytic study of edge betweenness in evolving scale-free and non-scale free trees. We aim at the probability distribution of edge betweenness under the condition that a local property, the in-degree of the "younger" node of a randomly selected edge, is known. En route to the conditional distribution of edge betweenness the exact joint distribution of cluster size and in-degree, and its one-dimensional marginal distributions have been presented in the paper as well. From the derived probability distributions the expectation values of different quantities have been calculated. Our results provide an exact solution not only for infinite, but for finite networks as well. PMID- 16711874 TI - Mean-field theory of localization in a fuse model. AB - We propose a mean-field theory for the localization of damage in a quasistatic fuse model on a cylinder. Depending on the quenched disorder distribution of the fuse thresholds, we show analytically that the system can either stay in a percolation regime up to breakdown, or start at some imposed current, to localize starting from the smallest scale (lattice spacing), or instead go to a diffuse localization regime where damage starts to concentrate in bands of width scaling as the width of the system, but remains diffuse at smaller scales. Depending on the nature of the quenched disorder on the fuse thresholds, we derive analytically the phase diagram of the system separating these regimes and the current levels for the onset of these possible localizations. We compare these predictions to numerical results. PMID- 16711875 TI - Levy-Brownian motion on finite intervals: Mean first passage time analysis. AB - We present the analysis of the first passage time problem on a finite interval for the generalized Wiener process that is driven by Levy stable noises. The complexity of the first passage time statistics (mean first passage time, cumulative first passage time distribution) is elucidated together with a discussion of the proper setup of corresponding boundary conditions that correctly yield the statistics of first passages for these non-Gaussian noises. The validity of the method is tested numerically and compared against analytical formulas when the stability index alpha approaches 2, recovering in this limit the standard results for the Fokker-Planck dynamics driven by Gaussian white noise. PMID- 16711876 TI - Rare events and the convergence of exponentially averaged work values. AB - Equilibrium free energy differences are given by exponential averages of nonequilibrium work values; such averages, however, often converge poorly, as they are dominated by rare realizations. I show that there is a simple and intuitively appealing description of these rare but dominant realizations. This description is expressed as a duality between "forward" and "reverse" processes, and provides both heuristic insights and quantitative estimates regarding the number of realizations needed for convergence of the exponential average. Analogous results apply to the equilibrium perturbation method of estimating free energy differences. The pedagogical example of a piston and gas [R.C. Lua and A.Y. Grosberg, J. Phys. Chem. B 109, 6805 (2005)] is used to illustrate the general discussion. PMID- 16711877 TI - Quantification of cross correlations in complex spatiotemporal systems. AB - We propose a design of the equal time correlation matrix suitable for the analysis of multivariate time series with ill-defined phases. We present the cross-correlation analysis of model data sets taken from coupled stochastic oscillators and compare the concept with the results obtained from a conventional correlation matrix analysis. We show that the concept provides a higher sensitivity combined with a better statistical significance when quantifying weak cross correlations. PMID- 16711878 TI - Microscopic features of moving traffic jams. AB - Empirical and numerical microscopic features of moving traffic jams are presented. Based on a single vehicle data analysis, it is found that within wide moving jams, i.e., between the upstream and downstream jam fronts there is a complex microscopic spatiotemporal structure. This jam structure consists of alternations of regions in which traffic flow is interrupted and flow states of low speeds associated with "moving blanks" within the jam. Moving blanks within a wide moving jam resemble electron holes in the valence band of semiconductors: As the moving blanks that propagate upstream appear due to downstream vehicle motion within the jam, so appearance of electron holes moving with the electric field results from electron motion against the electric field in the valence band of semiconductors. Empirical features of moving blanks are found. Based on microscopic models in the context of the Kerner's three-phase traffic theory, physical reasons for moving blanks emergence within wide moving jams are disclosed. Microscopic nonlinear effects of moving jam emergence, propagation, and dissolution as well as a diverse variety of hysteresis effects in freeway traffic associated with phase transitions and congested traffic propagation are numerically investigated. Microscopic structure of moving jam fronts is numerically studied and compared with empirical results. PMID- 16711879 TI - Efficient routing on complex networks. AB - We propose a routing strategy to improve the transportation efficiency on complex networks. Instead of using the routing strategy for shortest path, we give a generalized routing algorithm to find the so-called efficient path, which considers the possible congestion in the nodes along actual paths. Since the nodes with the largest degree are very susceptible to traffic congestion, an effective way to improve traffic and control congestion, as our strategy, can be redistributing traffic load in central nodes to other noncentral nodes. Simulation results indicate that the network capability in processing traffic is improved more than 10 times by optimizing the efficient path, which is in good agreement with the analysis. PMID- 16711880 TI - Scaling theory of temporal correlations and size-dependent fluctuations in the traded value of stocks. AB - Records of the traded value of fi stocks display fluctuation scaling, a proportionality between the standard deviation sigma(i) and the average : sigma(i) is proportional to alpha, with a strong time scale dependence alpha(Delta(t)). The nontrivial (i.e., neither 0.5 nor 1) value of alpha may have different origins and provides information about the microscopic dynamics. We present a set of stylized facts and then show their connection to such behavior. The functional form alpha(Delta(t)) originates from two aspects of the dynamics: Stocks of larger companies both tend to be traded in larger packages and also display stronger correlations of traded value. The results are integrated into a general framework that can be applied to a wide range of complex systems. PMID- 16711881 TI - Simulation estimates of cloud points of polydisperse fluids. AB - We describe two distinct approaches to obtaining the cloud-point densities and coexistence properties of polydisperse fluid mixtures by Monte Carlo simulation within the grand-canonical ensemble. The first method determines the chemical potential distribution mu(sigma) (with the polydisperse attribute) under the constraint that the ensemble average of the particle density distribution rho(sigma) match a prescribed parent form. Within the region of phase coexistence (delineated by the cloud curve) this leads to a distribution of the fluctuating overall particle density n, p(n), that necessarily has unequal peak weights in order to satisfy a generalized lever rule. A theoretical analysis shows that as a consequence, finite-size corrections to estimates of coexistence properties are power laws in the system size. The second method assigns mu(sigma) such that an equal-peak-weight criterion is satisfied for p(n) for all points within the coexistence region. However, since equal volumes of the coexisting phases cannot satisfy the lever rule for the prescribed parent, their relative contributions must be weighted appropriately when determining mu(sigma). We show how to ascertain the requisite weight factor operationally. A theoretical analysis of the second method suggests that it leads to finite-size corrections to estimates of coexistence properties which are exponentially small in the system size. The scaling predictions for both methods are tested via Monte Carlo simulations of a polydisperse lattice-gas model near its cloud curve, the results showing excellent quantitative agreement with the theory. PMID- 16711882 TI - Two-dimensional structure in a generic model of triangular proteins and protein trimers. AB - Motivated by the diversity and complexity of two-dimensional (2D) crystals formed by triangular proteins and protein trimers, we have investigated the structures and phase behavior of hard-disk trimers. In order to mimic specific binding interactions, each trimer possesses an "attractive" disk which can interact with similar disks on other trimers via an attractive square-well potential. At low density and low temperature, the fluid phase mainly consists of tetramers, pentamers, or hexamers. Hexamers provide the structural motif for a high-density, low-temperature periodic solid phase, but we also identify a metastable periodic structure based on a tetramer motif. At high density there is a transition between orientationally ordered and disordered solid phases. The connections between simulated structures and those of 2D protein crystals--as seen in electron microscopy--are briefly discussed. PMID- 16711883 TI - Local virial relation for self-gravitating system. AB - We demonstrate that the quasi-equilibrium state in a self-gravitating N-body system after cold collapse is uniquely characterized by the local virial relation using numerical simulations. Conversely, assuming the constant local virial ratio and Jeans equation for a spherically steady-state system, we investigate the full solution space of the problem under the constant anisotropy parameter and obtain some relevant solutions. Specifically, the local virial relation always provides a solution which has a power-law density profile in both the asymptotic regions r --> 0 and infinity. This type of solution is commonly observed in many numerical simulations. Only the anisotropic velocity dispersion controls this asymptotic behavior of density profile. PMID- 16711884 TI - Scale-free networks with an exponent less than two. AB - We study scale-free simple graphs with an exponent of the degree distribution gamma less than 2. Generically one expects such extremely skewed networks--which occur very frequently in systems of virtually or logically connected units--to have different properties than those of scale free networks with gamma>2: The number of links grows faster than the number of nodes and they naturally possess the small world property, because the diameter increases by the logarithm of the size of the network and the clustering coefficient is finite. We discuss a simple prototype model of such networks, inspired by real world phenomena, which exhibits these properties and allows for a detailed analytical investigation. PMID- 16711885 TI - Method for numerical simulation of two-term exponentially correlated colored noise. AB - A method for numerical simulation of two-term exponentially correlated colored noise is proposed. The method is an extension of traditional method for one-term exponentially correlated colored noise. The validity of the algorithm is tested by comparing numerical simulations with analytical results in two physical applications. PMID- 16711886 TI - Burridge-Knopoff model: Exploration of dynamic phases. AB - Slider-block models are often used to simulate earthquake dynamics. However, the models' origins are more conceptual than analytical. This study uses Navier's equations of an elastic bulk to derive a one-dimensional slider-block model, the Burridge-Knopoff model. This model exhibits a critical phase transition by varying the friction parameter. Accurate analytical estimates are made of event size limits for the small scale, large scale, and intermediate dynamic phases. The absence of large scale quasiperiodic delocalized events is noted for the parameter set investigated here. The time intervals between large scale events are approximately exponentially distributed for the system in its critical state, in agreement with the theory of nonequilibrium critical systems and earthquake dynamics. PMID- 16711887 TI - Diffusion on Archimedean lattices. AB - We consider random diffusive motion of classical particles over the edges of Archimedean lattices. The diffusion coefficient is obtained by using periodic orbit theory. We also study deterministic motion over a honeycomb lattice without the possibility for an immediate return to the preceding node, controlled by a tent map with the golden ratio slope. Numerical analysis is performed to confirm the theoretical results. PMID- 16711888 TI - Self-organized criticality and coevolution of network structure and dynamics. AB - We investigate, by numerical simulations, how the avalanche dynamics of the Bak Tang-Wiesenfeld sandpile model can induce emergence of scale-free networks and how this emerging structure affects dynamics of the system. PMID- 16711889 TI - Utterance selection model of language change. AB - We present a mathematical formulation of a theory of language change. The theory is evolutionary in nature and has close analogies with theories of population genetics. The mathematical structure we construct similarly has correspondences with the Fisher-Wright model of population genetics, but there are significant differences. The continuous time formulation of the model is expressed in terms of a Fokker-Planck equation. This equation is exactly soluble in the case of a single speaker and can be investigated analytically in the case of multiple speakers who communicate equally with all other speakers and give their utterances equal weight. Whilst the stationary properties of this system have much in common with the single-speaker case, time-dependent properties are richer. In the particular case where linguistic forms can become extinct, we find that the presence of many speakers causes a two-stage relaxation, the first being a common marginal distribution that persists for a long time as a consequence of ultimate extinction being due to rare fluctuations. PMID- 16711890 TI - Local versus global interactions in nonequilibrium transitions: A model of social dynamics. AB - A nonequilibrium system of locally interacting elements in a lattice with an absorbing order-disorder phase transition is studied under the effect of additional interacting fields. These fields are shown to produce interesting effects in the collective behavior of this system. Both for autonomous and external fields, disorder grows in the system when the probability of the elements to interact with the field is increased. There exists a threshold value of this probability beyond which the system is always disordered. The domain of parameters of the ordered regime is larger for nonuniform local fields than for spatially uniform fields. However, the zero field limit is discontinous. In the limit of vanishingly small probability of interaction with the field, autonomous or external fields are able to order a system that would fall in a disordered phase under local interactions of the elements alone. We consider different types of fields which are interpreted as forms of mass media acting on a social system in the context of Axelrod's model for cultural dissemination. PMID- 16711891 TI - Domain motion in the voter model with noise. AB - We study the voter model with noise on one-dimensional chains using Monte Carlo simulations and finite-size scaling techniques. We observe that the system evolution toward consensus is deeply affected by the addition of noise, and that the time to reach complete ordering increases with the noise parameter q. In particular, the simulations show that the average domain size scales as xi approximately q(-1/2) whereas the magnetization scales with the number of nodes as m approximately N(-1/2). PMID- 16711892 TI - Equilibrium statistics of an inelastically bouncing ball, subject to gravity and a random force. AB - We consider a particle moving on the half line x > 0 and subject to a constant force in the -x direction plus a delta-correlated random force. At x = 0 the particle is reflected inelastically. The velocities just after and before the reflection satisfy v(f) = -r v(i), where r is the coefficient of restitution. This simple model is of interest in connection with studies of driven granular matter in a gravitational field. With an exact analytical approach and simulations we study the steady-state distribution function P(x,v). PMID- 16711893 TI - Coarsening of topological defects in oscillating systems with quenched disorder. AB - We use large scale simulations to study interacting particles in two dimensions in the presence of both an ac drive and quenched disorder. As a function of ac amplitude, there is a crossover from a low drive regime where the colloid positions are highly disordered to a higher ac drive regime where the system dynamically reorders. We examine the coarsening of topological defects formed when the system is quenched from a disordered low ac amplitude state to a high ac amplitude state. When the quench is performed close to the disorder-order crossover, the defect density decays with time as a power law with alpha = 1/4 to 1/3. For deep quenches, in which the ac drive is increased to high values such that the dynamical shaking temperature is strongly reduced, we observe a logarithmic decay of the defect density into a grain boundary dominated state. We find a similar logarithmic decay of defect density in systems containing no pinning. We specifically demonstrate these effects for vortices in thin film superconductors, and discuss implications for dynamical reordering transition studies in these systems. PMID- 16711894 TI - Effect of shear on persistence in coarsening systems. AB - We analytically study the effect of a uniform shear flow on the persistence properties of coarsening systems. The study is carried out within the anisotropic Ohta-Jasnow-Kawasaki (OJK) approximation for a system with nonconserved scalar order parameter. We find that the persistence exponent theta has a nontrivial value: theta = 0.5034 in space dimension d = 3, and theta = 0.2406 for d = 2, the latter being exactly twice the value found for the unsheared system in d = 1. We also find that the autocorrelation exponent lambda is affected by shear in d = 3 but not in d = 2. PMID- 16711895 TI - Fluctuation-dissipation relation in an Ising model without detailed balance. AB - We consider the modified Ising model introduced by de Oliveira, Mendes, and Santos [J. Phys. A 26, 2317 (1993)], where the temperature depends locally on the spin configuration and detailed balance and local equilibrium are not obeyed. We derive a relation between the linear response function and correlation functions that generalizes the fluctuation-dissipation theorem. In the stationary states of the model, which are the counterparts of the Ising equilibrium states, the fluctuation-dissipation theorem breaks down due to the lack of time reversal invariance. In the nonstationary phase-ordering kinetics, the parametric plot of the integrated response function chi(t,t(w)) vs the autocorrelation function is different from that of the kinetic Ising model. However, splitting chi(t,t(w)) into a stationary and an aging term chi(t,t(w)) = chi(st)(t-t(w)) + chi(ag)(t,t(w)), we find chi(ag)(t,t(w)) approximately t(w)(-a(chi)) f(t/t(w)), and a numerical value of a(chi) consistent with a(chi)= 1/4, as in the kinetic Ising model. PMID- 16711896 TI - Nonequilibrium statistical physics with fictitious time. AB - Problems in nonequilibrium statistical physics are characterized by the absence of a fluctuation dissipation theorem. The usual analytic route for treating these vast class of problems is to use response fields in addition to the real fields that are pertinent to a given problem. This line of argument was introduced by Martin, Siggia, and Rose. We show that instead of using the response field, one can, following the stochastic quantization of Parisi and Wu, introduce a fictitious time. In this extra dimension a fluctuation dissipation theorem is built in and provides a different outlook to problems in nonequilibrium statistical physics. PMID- 16711897 TI - Equivalence of trans paths in ion channels. AB - We explore stochastic models for the study of ion transport in biological cells. Analysis of these models explains and explores an interesting feature of ion transport observed by biophysicists. Namely, the average time it takes ions to cross certain ion channels is the same in either direction, even if there is an electric potential difference across the channels. It is shown for simple single ion models that the distribution of a path (i.e., the history of location versus time) of an ion crossing the channel in one direction has the same distribution as the time-reversed path of an ion crossing the channel in the reverse direction. Therefore, not only is the mean duration of these paths equal, but other measures, such as the variance of passage time or the mean time a path spends within a specified section of the channel, are also the same for both directions of traversal. The feature is also explored for channels with interacting ions. If a system of interacting ions is in reversible equilibrium (net flux is zero), then the equivalence of the left-to-right trans paths with the time-reversed right-to-left trans paths still holds. However, if the system is in equilibrium, but not reversible equilibrium, then such equivalence need not hold. PMID- 16711898 TI - Wavy front dynamics in a three-component reaction-diffusion system with one activator and two inhibitors. AB - An analytic description for traveling waves in a one-dimensional reaction diffusion system with one activator and two inhibitors and with equal diffusion constants is developed using a piecewise linear approximation for the nonlinear activator reaction term. The case of front waves is examined in more detail, the monotonic and oscillating fronts being separately considered. The corresponding wave profiles are constructed, and the speed equation is obtained and discussed. It is found that the fronts in the three-component model propagate faster than the fronts in the two-component system. The front interaction is studied using numerical calculations. The results show that at head-on collisions two oscillating fronts produce a wavy domain, which spreads in space with time. PMID- 16711899 TI - Breaking of translational symmetry of a traveling planar impulse in a two dimensional two-variable reaction-diffusion model. AB - The stability of a planar impulse in rectangular spatial domains for a two variable excitable reaction-diffusion system is numerically studied. The dependence of the stability on the size of the domain perpendicular to the direction of the propagation of the impulse is shown. The instability results in asymptotic stable curved impulses or an asymptotic spatiotemporal structure, which is generated similarly to the one-dimensional backfiring phenomenon. PMID- 16711900 TI - Fluctuation theorems for quantum master equations. AB - A quantum fluctuation theorem for a driven quantum subsystem interacting with its environment is derived based solely on the assumption that its reduced density matrix obeys a closed evolution equation--i.e., a quantum master equation (QME). Quantum trajectories and their associated entropy, heat, and work appear naturally by transforming the QME to a time-dependent Liouville space basis that diagonalizes the instantaneous reduced density matrix of the subsystem. A quantum integral fluctuation theorem, a steady-state fluctuation theorem, and the Jarzynski relation are derived in a similar way as for classical stochastic dynamics. PMID- 16711901 TI - Temperature dependence of universal fluctuations in the two-dimensional harmonic XY model. AB - We compute exact analytical expressions for the skewness and kurtosis in the two dimensional harmonic XY model. These quantities correspond to the third and fourth normalized moments of the probability density function (PDF) of the magnetization of the model. From their behavior, we conclude that they depend explicitly on the system temperature even in the thermodynamic limit, and hence the PDF itself must depend on it. Our results correct the hypothesis called universal fluctuations, they confirm and extend previous results which showed a T dependence of the PDF, including perturbative expansions within the XY model up to first order in temperature. PMID- 16711902 TI - Epidemic variability in complex networks. AB - We study numerically the variability of the outbreak of diseases on complex networks. We use a susceptible-infected model to simulate the disease spreading at short times in homogeneous and in scale-free networks. In both cases, we study the effect of initial conditions on the epidemic dynamics and its variability. The results display a time regime during which the prevalence exhibits a large sensitivity to noise. We also investigate the dependence of the infection time of a node on its degree and its distance to the seed. In particular, we show that the infection time of hubs have non-negligible fluctuations which limit their reliability as early detection stations. Finally, we discuss the effect of the multiplicity of paths between two nodes on the infection time. In particular, we demonstrate that the existence of even long paths reduces the average infection time. These different results could be of use for the design of time-dependent containment strategies. PMID- 16711903 TI - Fluctuation-dissipation relations in driven dissipative systems. AB - Exact theoretical results for the violation of time-dependent fluctuation dissipation relations in driven dissipative systems are presented. The ratio of the correlation to delayed response in the stochastic model introduced in [Phys. Rev. Lett. 93, 240601 (2004)] is shown to depend on measurement time. The fluctuation temperature defined by this ratio differs both from the temperature of the environment performing the driving, and from other effective temperatures of the system, such as the average energy (or "granular temperature"). General explanations are given for the time independence of the fluctuation temperature for simple measurements or long measurement times. PMID- 16711904 TI - Fractional Fokker-Planck dynamics: Numerical algorithm and simulations. AB - Anomalous transport in a tilted periodic potential is investigated numerically within the framework of the fractional Fokker-Planck dynamics via the underlying continuous-time random walk. An efficient numerical algorithm is developed which is applicable for an arbitrary potential. This algorithm is then applied to investigate the fractional current and the corresponding nonlinear mobility in different washboard potentials. Normal and fractional diffusion are compared through their time evolution of the probability density in state space. Moreover, we discuss the stationary probability density of the fractional current values. PMID- 16711905 TI - Properties of the probability distribution associated with the largest event in an earthquake cluster and their implications to foreshocks. AB - The space-time epidemic-type aftershock sequence model is a stochastic branching process in which earthquake activity is classified into background and clustering components and each earthquake triggers other earthquakes independently according to certain rules. This paper gives the probability distributions associated with the largest event in a cluster and their properties for all three cases when the process is subcritical, critical, and supercritical. One of the direct uses of these probability distributions is to evaluate the probability of an earthquake to be a foreshock, and magnitude distributions of foreshocks and nonforeshock earthquakes. To verify these theoretical results, the Japan Meteorological Agency earthquake catalog is analyzed. The proportion of events that have 1 or more larger descendants in total events is found to be as high as about 15%. When the differences between background events and triggered event in the behavior of triggering children are considered, a background event has a probability about 8% to be a foreshock. This probability decreases when the magnitude of the background event increases. These results, obtained from a complicated clustering model, where the characteristics of background events and triggered events are different, are consistent with the results obtained in [Ogata, Geophys. J. Int. 127, 17 (1996)] by using the conventional single-linked cluster declustering method. PMID- 16711906 TI - Exact finite-size scaling functions for the interfacial tensions of the Ising model on planar lattices. AB - Exact finite-size scaling functions of the interfacial tensions are obtained for the Ising model with isotropic coupling on a set of M x N planar lattices, including square (sq), plane triangular (pt), and honeycomb (hc) lattices. The analyses of transitive behaviors at criticality revise the knowledge of the interfacial tensions as a function of the aspect ratio defined by R = M/N for R approaching to zero gradually. The amplitudes of the interfacial tensions for the sq, pt, and hc lattices are further shown to have relative proportions 1:square root of 3 / 2:square root of 3 which are related to the aspect ratios for the three lattices to have similar domains. PMID- 16711907 TI - Renewal, modulation, and superstatistics in times series. AB - We consider two different approaches, to which we refer to as renewal and modulation, to generate time series with a nonexponential distribution of waiting times. We show that different time series with the same waiting time distribution are not necessarily statistically equivalent, and might generate different physical properties. Renewal generates aging and anomalous scaling, while modulation yields no significant aging and either ordinary or anomalous diffusion, according to the dynamic prescription adopted. We show, in fact, that the physical realization of modulation generates two classes of events. The events of the first class are determined by the persistent use of the same exponential time scale for an extended lapse of time, and consequently are numerous; the events of the second class are identified with the abrupt changes from one to another exponential prescription, and consequently are rare. The events of the second class, although rare, determine the scaling of the diffusion process, and for this reason we term them as crucial events. According to the prescription adopted to produce modulation, the distribution density of the time distances between two consecutive crucial events might have, or not, a diverging second moment. In the former case the resulting diffusion process, although going through a transition regime very extended in time, will eventually become anomalous. In conclusion, modulation rather than ruling out the action of renewal events, produces crucial events hidden by clouds of exponential events, thereby setting the challenge for their identification. PMID- 16711908 TI - Ordering spatiotemporal chaos in complex thermosensitive neuron networks. AB - We have studied the effect of random long-range connections in chaotic thermosensitive neuron networks with each neuron being capable of exhibiting diverse bursting behaviors, and found stochastic synchronization and optimal spatiotemporal patterns. For a given coupling strength, the chaotic burst-firings of the neurons become more and more synchronized as the number of random connections (or randomness) is increased and, rather, the most pronounced spatiotemporal pattern appears for an optimal randomness. As the coupling strength is increased, the optimal randomness shifts towards a smaller strength. This result shows that random long-range connections can tame the chaos in the neural networks and make the neurons more effectively reach synchronization. Since the model studied can be used to account for hypothalamic neurons of dogfish, catfish, etc., this result may reflect the significant role of random connections in transferring biological information. PMID- 16711909 TI - Efficiency of information spreading in a population of diffusing agents. AB - We introduce a model for information spreading among a population of N agents diffusing on a square L x L lattice, starting from an informed agent (Source). Information passing from informed to unaware agents occurs whenever the relative distance is < or = 1. Numerical simulations show that the time required for the information to reach all agents scales as N(-alpha)L(beta), where alpha and beta are noninteger. A decay factor z takes into account the degeneration of information as it passes from one agent to another; the final average degree of information of the population tau(av)(z) is thus history dependent. We find that the behavior of tau(av)(z) is nonmonotonic with respect to N and L and displays a set of minima. Part of the results are recovered with analytical approximations. PMID- 16711910 TI - Cultural propagation on social networks. AB - In this work we present a model for the propagation of culture on networks of different topology and by considering different underlying dynamics. We extend a previous model proposed by Axelrod by letting a majority govern the dynamics of changes. This in turn allows us to define a Lyapunov functional for the system. PMID- 16711911 TI - Duality between quantum and classical dynamics for integrable billiards. AB - We establish a duality between the quantum wave vector spectrum and the eigenmodes of the classical Liouvillian dynamics for integrable billiards. Signatures of the classical eigenmodes appear as peaks in the correlation function of the quantum wave vector spectrum. A semiclassical derivation and numerical calculations are presented in support of the results. These classical eigenmodes can be observed in physical experiments through the autocorrelation of the transmission coefficient of waves in quantum billiards. Exact classical trace formulas of the resolvent are derived for the rectangle, equilateral triangle, and circle billiards. We also establish a correspondence between the classical periodic orbit length spectrum and the quantum spectrum for integrable polygonal billiards. PMID- 16711912 TI - Dynamical singularities for complex initial conditions and the motion at a real separatrix. AB - This work investigates singularities occurring at finite real times in the classical dynamics of one-dimensional double-well systems with complex initial conditions. The objective is to understand the relationship between these singularities and the behavior of the systems for real initial conditions. An analytical treatment establishes that the dynamics of a quartic double well system possesses a doubly infinite sequence of singularities. These are associated with initial conditions that converge to those for the real separatrix as the singularity time becomes infinite. This confluence of singularities is shown to lead to the unstable behavior that characterizes the real motion at the separatrix. Numerical calculations confirm the existence of a large number of singularities converging to the separatrix for this and two additional double well systems. The approach of singularities to the real axis is of particular interest since such behavior has been related to the formation of chaos in nonintegrable systems. The properties of the singular trajectories which cause this convergence to the separatrix are identified. The hyperbolic fixed point corresponding to the potential energy maximum, responsible for the characteristic motion at a separatrix, also plays a critical role in the formation of the complex singularities by delaying trajectories and then deflecting them into asymptotic regions of space from where they are directly repelled to infinity in a finite time. PMID- 16711913 TI - Method for measuring unstable dimension variability from time series. AB - Many of the results in the theory of dynamical systems rely on the assumption of hyperbolicity. One of the possible violations of this condition is the presence of unstable dimension variability (UDV), i.e., the existence in a chaotic attractor of sets of unstable periodic orbits, each with a different number of expanding directions. It has been shown that the presence of UDV poses severe limitations to the length of time for which a numerically generated orbit can be assumed to lie close to a true trajectory of such systems (the shadowing time). In this work we propose a method to detect the presence of UDV in real systems from time series measurements. Variations in the number of expanding directions are detected by determining the local topological dimension of the unstable space for points along a trajectory on the attractor. We show for a physical system of coupled electronic oscillators that with this method it is possible to decompose attractors into subsets with different unstable dimension and from this gain insight into the times a typical trajectory spends in each region. PMID- 16711914 TI - Dephasing representation of quantum fidelity for general pure and mixed states. AB - A general semiclassical expression for quantum fidelity (Loschmidt echo) of arbitrary pure and mixed states is derived. It expresses fidelity as an interference sum of dephasing trajectories weighed by the Wigner function of the initial state, and does not require that the initial state be localized in position or momentum. This general dephasing representation is special in that, counterintuitively, all of fidelity decay is due to dephasing and none is due to the decay of classical overlaps. Surprising accuracy of the approximation is justified by invoking the shadowing theorem: twice--both for physical perturbations and for numerical errors. Beyond justifying the approximation, the shadowing theorem makes the dephasing representation practical: without shadowing it would be impossible to find numerically the precise trajectories needed in a semiclassical approximation. It is shown how the general expression reduces to the previously known special forms for localized states. The superiority of the general over the specialized forms is explained and supported by numerical tests for wave packets, nonlocal pure states, and for simple and random mixed states. The tests are done in nonuniversal regimes in mixed phase space where detailed features of fidelity are important. Although semiclassically motivated, the present approach is valid for abstract systems with a finite Hilbert basis provided that the discrete Wigner transform is used. This makes the method applicable, via a phase-space approach, to problems of quantum computation. PMID- 16711915 TI - Semiconductor laser under resonant feedback from a Fabry-Perot resonator: Stability of continuous-wave operation. AB - We study the continuous-wave (cw) operation of a semiconductor laser subject to optical feedback from a Fabry-Perot resonator in a case where the emission is resonant to a reflection minimum of the resonator. This configuration is treated in the framework of Lang-Kobayashi equations. The nature of bifurcations and the stability of steady state solutions is analyzed in terms of the dependence on magnitude and phase of the feedback. In contrast to conventional optical feedback from a single mirror, the locus of external cavity modes is not elliptic but represents a tilted eight with possible satellite bubbles. Below a critical feedback strength, which is analytically given, only one single mode exists representing the completely unchanged cw emission of the laser. In this weak feedback regime, the feedback phase allows noninvasive control of the cw emission and a tailoring of its small-signal response within wide limits. The results obtained are a prototype for all-optical realizations of delayed feedback control. PMID- 16711916 TI - Chaotic properties of planar elongational flow and planar shear flow: Lyapunov exponents, conjugate-pairing rule, and phase space contraction. AB - The simulation of planar elongational flow in a nonequilibrium steady state for arbitrarily long times has recently been made possible, combining the SLLOD algorithm with periodic boundary conditions for the simulation box. We address the fundamental questions regarding the chaotic behavior of this type of flow, comparing its chaotic properties with those of the well-established SLLOD algorithm for planar shear flow. The spectra of Lyapunov exponents are analyzed for a number of state points where the energy dissipation is the same for both flows, simulating a nonequilibrium steady state for isoenergetic and isokinetic constrained dynamics. We test the conjugate-pairing rule and confirm its validity for planar elongation flow, as is expected from the Hamiltonian nature of the adiabatic equations of motion. Remarks about the chaoticity of the convective part of the flows, the link between Lyapunov exponents and viscosity, and phase space contraction for both flows complete the study. PMID- 16711917 TI - Quantum-classical transition of the open quartic oscillator: The role of the environment. AB - We investigate the classical and quantum dynamics of the open quartic oscillator model. Typically quantum behavior such as collapses and revivals (also squeezing) are induced by the nonlinearity of the model. We show that purely diffusive environments, as expected, attenuate such phenomena. We obtain analytical results in both regimes classical and quantum and discuss the effect of a diffusive reservoir in the two cases. We show that "separation times" as usually defined in the literature are strongly observable (and initial condition) dependent, rendering a solid definition of a unique classical limit rather difficult. In particular, the separation time for the variance can be smaller than that for the expectation value of the position of the centroid of the wave packet. We find a hierarchy of time scales which depends on the observable and the reservoir. PMID- 16711918 TI - Characterization of fluctuations of impedance and scattering matrices in wave chaotic scattering. AB - In wave chaotic scattering, statistical fluctuations of the scattering matrix S and the impedance matrix Z depend both on universal properties and on nonuniversal details of how the scatterer is coupled to external channels. This paper considers the impedance and scattering variance ratios, Xi(z) and Xi(s), where Xi(z) = Var[Z(ij)]/{Var[Z(ii)]Var[Z(jj)]}1/2, Xi(s) = Var[S(ij)]/{Var[S(ii)]Var[S(jj)]}1/2, and Var[.] denotes variance. Xi(z) is shown to be a universal function of distributed losses within the scatterer. That is, Xi(z) is independent of nonuniversal coupling details. This contrasts with s for which universality applies only in the large loss limit. Explicit results are given for Xi(z) for time reversal symmetric and broken time reversal symmetric systems. Experimental tests of the theory are presented using data taken from scattering measurements on a chaotic microwave cavity. PMID- 16711919 TI - Domain chaos puzzle and the calculation of the structure factor and its half width. AB - The disagreement of the scaling of the correlation length xi between experiment and the Ginzburg-Landau (GL) model for domain chaos was resolved. The Swift Hohenberg (SH) domain chaos model was integrated numerically to acquire test images to study the effect of a finite image size on the extraction of from the structure factor (SF). The finite image size had a significant effect on the SF determined with the Fourier-transform (FT) method. The maximum entropy method (MEM) was able to overcome this finite image-size problem and produced fairly accurate SFs for the relatively small image sizes provided by experiments. Correlation lengths often have been determined from the second moment of the SF of chaotic patterns because the functional form of the SF is not known. Integration of several test functions provided analytic results indicating that this may not be a reliable method of extracting xi. For both a Gaussian and a squared SH form, the correlation length xi[triple bond]1/sigma, determined from the variance sigma2 of the SF, has the same dependence on the control parameter epsilon as the length xi contained explicitly in the functional forms. However, for the SH and the Lorentzian forms we find xi approximately xi (1/2). Results for xi determined from new experimental data by fitting the functional forms directly to the experimental SF yielded xi approximately epsilon(-v) with v approximately equal to 1/4 for all four functions in the case of the FT method, but v approximately equal to 1/2, in agreement with the GL prediction, in the case of the MEM. Over a wide range of epsilon and wave number k, the experimental SFs collapsed onto a unique curve when appropriately scaled by xi. PMID- 16711920 TI - Effect of noise on generalized chaotic synchronization. AB - When two characteristically different chaotic oscillators are coupled, generalized synchronization can occur. Motivated by the phenomena that common noise can induce and enhance complete synchronization or phase synchronization in chaotic systems, we investigate the effect of noise on generalized chaotic synchronization. We develop a phase-space analysis, which suggests that the effect can be system dependent in that common noise can either induce/enhance or destroy generalized synchronization. A prototype model consisting of a Lorenz oscillator coupled with a dynamo system is used to illustrate these phenomena. PMID- 16711921 TI - Quasiperiodic and chaotic discrete breathers in a parametrically driven system without linear dispersion. AB - We study a one-dimensional lattice of anharmonic oscillators with only quartic nearest-neighbor interactions, in which discrete breathers (DB's) can be explicitly constructed by an exact separation of their time and space dependence. Introducing parametric periodic driving, we first show how a variety of such DB's can be obtained by selecting spatial profiles from the homoclinic orbits of an invertible map and combining them with initial conditions chosen from the Poincare surface of section of a simple Duffing's equation. Placing then our initial conditions at the center of the islands of a major resonance, we demonstrate how the corresponding DB can be stabilized by varying the amplitude of the driving. We thus discover around elliptic points a large region of quasiperiodic breathers, which are stable for very long times. Starting with initial conditions close to the elliptic point at the origin, we find that as we approach the main chaotic layer, a quasiperiodic breather either destabilizes by delocalization or turns into a chaotic breather, with an evidently broadbanded Fourier spectrum before it collapses. For some breather profiles stable quasiperiodic breathers exist all the way to the separatrix of the Duffing equation, indicating the presence of large regions of tori around the DB solution in the multidimensional phase space. We argue that these strong localization phenomena are due to the absence of phonon resonances, as there are no linear dispersion terms in our lattices. We also show, however, that these phenomena persist in more realistic physical models, in which weak linear dispersion is included in the equations of motion, with a sufficiently small coefficient. PMID- 16711922 TI - Feedback control of subcritical oscillatory instabilities. AB - Feedback control of a subcritical oscillatory instability is investigated in the framework of a globally-controlled complex Ginzburg-Landau equation that describes the nonlinear dynamics near the instability threshold. The control is based on a feedback loop between the system linear growth rate and the maximum of the amplitude of the emerging pattern. It is shown that such control can suppress the blow up and result in the formation of spatially localized pulses similar to oscillons. In the one-dimensional case, depending on the values of the linear and nonlinear dispersion coefficients, several types of the pulse dynamics are possible in which the computational domain contains: (i) a single stationary pulse; (ii) several coexisting stationary pulses; (iii) competing pulses that appear one after another at random locations so that at each moment of time there is only one pulse in the domain; (iv) spatiotemporally chaotic system of short pulses; (v) spatially-synchronized pulses. Similar dynamic behavior is found also in the two-dimensional case. The effect of the feedback delay is also studied. It is shown that the increase of the delay leads to an oscillatory instability of the pulses and the formation of pulses with oscillating amplitude. PMID- 16711923 TI - Statistics of pressure fluctuations in decaying isotropic turbulence. AB - We present results from a systematic direct-numerical simulation study of pressure fluctuations in an unforced, incompressible, homogeneous, and isotropic three-dimensional turbulent fluid. At cascade completion, isosurfaces of low pressure are found to be organized as slender filaments, whereas the predominant isostructures appear sheetlike. We exhibit several results, including plots of probability distributions of the spatial pressure difference, the pressure gradient norm, and the eigenvalues of the pressure-Hessian tensor. Plots of the temporal evolution of the mean pressure-gradient norm, and the mean eigenvalues of the pressure-Hessian tensor are also exhibited. We find the statistically preferred orientations between the eigenvectors of the pressure-Hessian tensor, the pressure gradient, the eigenvectors of the strain-rate tensor, the vorticity, and the velocity. Statistical properties of the nonlocal part of the pressure Hessian tensor are also exhibited. We present numerical tests (in the viscous case) of some conjectures of Ohkitani [Phys. Fluids A 5, 2570 (1993)] and Ohkitani and Kishiba [Phys. Fluids 7, 411 (1995)] concerning the pressure-Hessian and the strain-rate tensors, for the unforced, incompressible, three-dimensional Euler equations. PMID- 16711924 TI - Fluctuations in Saffman-Taylor fingers with quenched disorder. AB - We make an experimental characterization of the effect that static disorder has on the shape of a normal Saffman-Taylor finger. We find that static noise induces a small amplitude and long wavelength instability on the sides of the finger. Fluctuations on the finger sides have a dominant wavelength, indicating that the system acts as a selective amplifier of static noise. The dominant wavelength does not seem to be very sensitive to the intensity of static noise present in the system. On the other hand, at a given flow rate, rms fluctuations of the finger width, decrease with decreasing intensity of static noise. This might explain why the sides of the fingers are flat for typical Saffman-Taylor experiments. Comparison with previous numerical studies of the effect that temporal noise has on the Saffman-Taylor finger, leads to conclude that the effect of temporal noise and static noise are similar. The behavior of fluctuations of the finger width found in our experiments, is qualitatively similar to one recently reported, in the sense that, the magnitude of the width fluctuations decays as a power law of the capillary number, at low flow rates, and increases with capillary number for larger flow rates. PMID- 16711925 TI - Effect of heat flux on differential rotation in turbulent convection. AB - We studied the effect of the turbulent heat flux on the Reynolds stresses in a rotating turbulent convection. To this end we solved a coupled system of dynamical equations which includes the equations for the Reynolds stresses, the entropy fluctuations, and the turbulent heat flux. We used a spectral tau approximation in order to close the system of dynamical equations. We found that the ratio of the contributions to the Reynolds stresses caused by the turbulent heat flux and the anisotropic eddy viscosity is of the order of approximately 10(L rho/l0)2, where l0 is the maximum scale of turbulent motions and L rho is the fluid density variation scale. This effect is crucial for the formation of the differential rotation and should be taken into account in the theories of the differential rotation of the Sun, stars, and planets. In particular, we demonstrated that this effect may cause the differential rotation which is comparable with the typical solar differential rotation. PMID- 16711926 TI - Bifurcation analysis of multiple steady flow patterns for Rayleigh-Benard convection in a cubical cavity at Pr = 130. AB - The bifurcation diagram of steady convective flow patterns inside a cubical cavity with adiabatic lateral walls heated from below and filled with silicone oil (Pr = 130) was determined for values of the Rayleigh number (Ra) up to 1.5 x 10(5). A continuation procedure based on the Galerkin spectral method was used to determine the steady convective solutions as a function of Ra. Bifurcations leading to either new steady or time-dependent solutions were identified and new steady solution branches were also continued. A total of fifteen steady solutions were tracked and the stability analysis predicted that six flow patterns were stable and that two, three, or even four of these patterns coexisted over certain ranges of Ra in the studied domain. Predicted flow patterns and transitions are in agreement with flow visualizations previously reported in the literature. The variation of the Nusselt number (Nu) as a function of Pr was investigated for three of the stable flow patterns identified: a x or y roll, a diagonal oriented roll and a pattern formed by four connected half rolls. It was found that whereas the Nusselt changes within the region 0.71 < or = Pr < or = 10 it tends to an asymptotic value with increasing Pr. PMID- 16711927 TI - Enhanced Faraday pattern stability with three-frequency driving. AB - We report experimental observations of enhanced stability of quasipatterns and superlattice patterns in a vertically oscillated, deep viscous fluid layer with the addition of a third driving frequency. With two-frequency driving in the ratios 4:5 and 6:7, 12-fold quasipatterns and type-I superlattice patterns appear, respectively, as a secondary instability for a range of relative phases and amplitudes. Addition of a small third-frequency component at twice the difference frequency, i.e., 4:5:2 and 6:7:2, shifts the region of stability for these patterns closer to onset. For a range of parameter values the stabilized patterns become the primary instability. The degree of stabilization is sensitive to the amplitude and relative phase of the third-frequency term in qualitative agreement with a recent symmetry based analysis of resonant three-wave interactions. PMID- 16711928 TI - Numerical study of pattern formation in miscible rotating Hele-Shaw flows. AB - The dynamics of the diffusing interface separating two miscible fluids in a rotating Hele-Shaw cell is studied by intensive and highly accurate numerical simulations. We perform numerical experiments in a wide range of parameters, focusing on the influence of viscosity contrast and Korteweg stresses on the shape of the interfacial patterns. A great variety of morphological behaviors is systematically introduced, and a wealth of interesting phenomena related to finger competition dynamics, filament stretching, and interface pinch off are reveal. Our simulations exhibit miscible patterns that bear a strong resemblance to their immiscible counterparts for larger Korteweg stresses. The quantitative equivalence between such stresses and the usual immiscible surface tension is studied. The concept of an effective interfacial tension is considered, allowing the direct and precise calculation of the important fingering properties under miscible circumstances. Our results show excellent agreement with existing experiments and simulations for corresponding immiscible displacements. This agreement refers to a striking similarity between miscible and immiscible pattern morphologies, and also to an accurate prediction for the typical number of miscible fingering structures formed. Our findings suggest that the effective interfacial tension is both qualitatively and quantitatively equivalent to its immiscible counterpart. PMID- 16711929 TI - Robust control for convection suppression in a fluid layer: The effects of boundary properties, actuator lag, and major parameter uncertainties. AB - The robustness of control is a requirement to maintain a fluid layer at conductive equilibrium heated to a highly supercritical condition. Robustness determines how much uncertainties, or design parameter mismatches, can be tolerated. Both linear stability analysis and three-dimensional fully nonlinear simulations are used for the study of the linear quadratic Gaussian (LQG) controller. The parameter mismatches from the nominal conditions are introduced into the plant model, while the LQG compensator assumes nominal conditions. The mismatches arise from boundary properties, actuator lag, sensor level uncertainty, and wall thickness, as well as from the major parameters such as Prandtl number, Rayleigh number, wave number, and truncation number in the reduced-order model. The results suggest that the LQG compensator action can preserve closed-loop stability at over ten times the critical Rayleigh number, provided that the mismatches in the sensor level and wall thickness are small. Mismatches in the Prandtl number and wall material properties have little impact. Mismatches in Rayleigh number and wave number are relatively benign compared with the sensor and thickness parameters. Techniques for measuring the plant output temperature at multiple levels with sufficient accuracy may be an implementation challenge. PMID- 16711930 TI - Dynamics and thermodynamics of axisymmetric flows: Theory. AB - We develop variational principles to study the structure and the stability of equilibrium states of axisymmetric flows. We show that the axisymmetric Euler equations for inviscid flows admit an infinite number of steady state solutions. We find their general form and provide analytical solutions in some special cases. The system can be trapped in one of these steady states as a result of an inviscid violent relaxation. We show that the stable steady states maximize a (nonuniversal) function while conserving energy, helicity, circulation, and angular momentum (robust constraints). This can be viewed as a form of generalized selective decay principle. We derive relaxation equations which can be used as numerical algorithm to construct nonlinearly dynamically stable stationary solutions of axisymmetric flows. We also develop a thermodynamical approach to predict the equilibrium state at some fixed coarse-grained scale. We show that the resulting distribution can be divided in two parts: one universal coming from the conservation of robust invariants and one non-universal determined by the initial conditions through the fragile invariants (for freely evolving systems) or by a prior distribution encoding nonideal effects such as viscosity, small-scale forcing, and dissipation (for forced systems). Finally, we derive a parametrization of inviscid mixing to describe the dynamics of the system at the coarse-grained scale. A conceptual interest of this axisymmetric model is to be intermediate between two-dimensional (2D) and 3D turbulence. PMID- 16711931 TI - Equilibria for the relative motion of three heavy spheres in Stokes fluid flow. AB - Dynamics of three identical solid spheres falling under gravity in low-Reynolds number fluid flow is investigated. Stationary solutions are found. Their stability is discussed. Phase portraits for two types of symmetric motions are calculated. PMID- 16711932 TI - Induction, helicity, and alpha effect in a toroidal screw flow of liquid gallium. AB - We investigate experimentally induction mechanisms in a screw flow of gallium in a toroidal channel. The flow is nonstationary and operated in a spin-down regime: the channel (and fluid) are initially set into solid body rotation; as the channel is stopped the fluid is set into strong helical motion by diverters located inside the channel. In this study, we put a particular emphasis on the induction generated by these helical motions, which are expected to develop over the entire range of turbulent scales. We apply an external magnetic field either perpendicular to the channel axis parallel to it. At large scales the nonlinear induction mechanisms are associated with the Parker stretch and twist effect and with the expulsion due to overall rotation. Induction mechanisms can also originate in the small scale helicity as in the alpha induction effect of mean field magnetohydrodynamics. Our measurements yield an upper bound for the alpha coefficient, significantly lower than estimates based on dimensional analysis. We discuss the consequences of our observations for the engineering of homogeneous dynamos in the laboratory. PMID- 16711933 TI - Ion energy distribution and gas heating in the cathode fall of a direct-current microdischarge. AB - This paper reports on measurements of the ion energy distribution (IED) at the cathode of an argon dc microdischarge using energy-resolved molecular beam mass spectrometry. The measurements are conducted at a fixed pressure-electrode separation product (pd) of 1 cm Torr with a maximum discharge pressure of 20 Torr. The measured IED is compared to the theory of Davis and Vanderslice [W. D. Davis and T. A. Vanderslice, Phys. Rev. 131, 219 (1963)]. A higher pressure in a case of almost constant normalized current densities by pressure (Jp(-2) = 0.080+/-0.006 mA/cm(-2) Torr(-2)) yields a lower ratio of the ion mean free path to the sheath thickness. The results in almost constant Jp(-2) case then indicate that a scaling law of Jp(-2) is no longer applicable for IED of microdischarge. Expected background gaseous temperatures from IEDs with the collisional Child law have reasonable increasing with increased current density (J) in both cases of almost constant Jp(-2) and a constant pressure of 10 Torr. Supported by temperature measurement by laser absorption spectroscopy, it is demonstrated that the expanded theory might be applicable also to microdischarges (Ar approximately 20 Torr) with temperature adjusting. PMID- 16711934 TI - Fast-electron transport and heating of solid targets in high-intensity laser interactions measured by K alpha fluorescence. AB - We present experimental results on fast-electron energy deposition into solid targets in ultrahigh intensity laser-matter interaction. X-ray K alpha emission spectroscopy with absolute photon counting served to diagnose fast-electron propagation in multilayered targets. Target heating was measured from ionization shifted K alpha emission. Data show a 200 microm fast-electron range in solid Al. The relative intensities of spectrally shifted Al K alpha lines imply a mean temperature of a few tens of eV up to a 100 microm depth. Experimental results suggest refluxing of the electron beam at target rear side. They were compared with the predictions of both a collisional Monte Carlo and a collisional electromagnetic, particle-fluid transport code. The validity of the code modeling of heating in such highly transient conditions is discussed. PMID- 16711935 TI - Nonlinear evolution of the plasma beat wave: Compressing the laser beat notes via electromagnetic cascading. AB - The near-resonant beat wave excitation of an electron plasma wave (EPW) can be employed for generating the trains of few-femtosecond electromagnetic (EM) pulses in rarefied plasmas. The EPW produces a comoving index grating that induces a laser phase modulation at the difference frequency. As a result, the cascade of sidebands red and blue shifted by integer multiples of the beat frequency is generated in the laser spectrum. The bandwidth of the phase-modulated laser is proportional to the product of the plasma length, laser wavelength, and amplitude of the electron density perturbation. When the beat frequency is lower than the electron plasma frequency, the redshifted spectral components are advanced in time with respect to the blueshifted ones near the center of each laser beat note. The group velocity dispersion of plasma compresses so chirped beat notes to a few-laser-cycle duration thus creating a train of sharp EM spikes with the beat periodicity. Depending on the plasma and laser parameters, chirping and compression can be implemented either concurrently in the same, or sequentially in different plasmas. Evolution of the laser beat wave and electron density perturbations is described in time and one spatial dimension in a weakly relativistic approximation. Using the compression effect, we demonstrate that the relativistic bistability regime of the EPW excitation [G. Shvets, Phys. Rev. Lett. 93, 195004 (2004)] can be achieved with the initially subthreshold beat wave pulse. PMID- 16711936 TI - Localized gap-edge fields of one-dimensional photonic crystals with an epsilon negative and a mu-negative defect. AB - We study the properties of one-dimensional photonic crystals with an epsilon negative and a mu-negative defect. With suitable parameters, the pair defect is equivalent to a transparent material with zero effective refractive index. This special pair defect has no influence on the spectral gap formed by the interference of propagating waves in positive-refractive-index materials. However, the field distribution is modified noticeably by the decaying wave in the pair defect. Particularly, the gap-edge field can be a highly localized wave instead of the usual standing wave as the size of the pair defect increases. The localized gap-edge field can reduce the switching thresholds for bistability greatly when Kerr-type nonlinear mu-negative material is involved. A nonideal model when the epsilon-negative and mu-negative materials are dispersive and lossy is used to verify the unusual properties. PMID- 16711937 TI - Discrete soliton mobility in two-dimensional waveguide arrays with saturable nonlinearity. AB - We address the issue of mobility of localized modes in two-dimensional nonlinear Schrodinger lattices with saturable nonlinearity. This describes, e.g., discrete spatial solitons in a tight-binding approximation of two-dimensional optical waveguide arrays made from photorefractive crystals. We discuss the numerically obtained exact stationary solutions and their stability, focusing on three different solution families with peaks at one, two, and four neighboring sites, respectively. When varying the power, there is a repeated exchange of stability between these three solutions, with symmetry-broken families of connecting intermediate stationary solutions appearing at the bifurcation points. When the nonlinearity parameter is not too large, we observe good mobility and a well defined Peierls-Nabarro barrier measuring the minimum energy necessary for rendering a stable stationary solution mobile. PMID- 16711938 TI - Governing soliton splitting in one-dimensional lattices. AB - We investigate discrete light dynamics in the presence of a longitudinal defect of arbitrary extension, amplitude and position in a nonlinear waveguide array. We model and discuss the physics of the soliton-defect interaction, showing how to gain complete control over the system outcome for soliton-based data processing. We propose all-optical management in dye-doped liquid crystals. PMID- 16711939 TI - Spontaneous curvature cancellation in forced thin sheets. AB - In this paper we report numerically observed spontaneous vanishing of mean curvature on a developable cone made by pushing a thin elastic sheet into a circular container [E. Cerda, S. Chaieb, F. Melo, and L. Mahadevan, Nature (London) 401 46 (1999)]. We show that this feature is independent of thickness of the sheet, the supporting radius, and the amount of deflection. Several variants of the developable cone are studied to examine the necessary conditions that lead to the vanishing of mean curvature. It is found that the presence of appropriate amount of radial stress is necessary. The developable cone geometry somehow produces the right amount of radial stress to induce just enough radial curvature to cancel the conical azimuthal curvature. In addition, the circular symmetry of supporting container edge plays an important role. With an elliptical supporting edge, the radial curvature overcompensates the azimuthal curvature near the minor axis and undercompensates near the major axis. Our numerical finding is verified by a crude experiment using a reflective plastic sheet. We expect this finding to have broad importance in describing the general geometrical properties of forced crumpling of thin sheets. PMID- 16711940 TI - Inhomogeneous soliton ratchets under two ac forces. AB - We extend our previous work on soliton ratchet devices [L. Morales-Molina, Eur. Phys. J. B 37, 79 (2004)] to consider the joint effect of two ac forces including nonharmonic drivings, as proposed for particle ratchets by Savele'v [Europhys. Lett. 67, 179 (2004); Phys. Rev. E 70, 066109 (2004)]. Current reversals due to the interplay between the phases, frequencies, and amplitudes of the harmonics are obtained. An analysis of the effect of the damping coefficient on the dynamics is presented. We show that solitons give rise to nontrivial differences in the phenomenology reported for particle systems that arise from their extended character. A comparison with soliton ratchets in homogeneous systems with biharmonic forces is also presented. This ratchet device may be an ideal candidate for Josephson junction ratchets with intrinsic large damping. PMID- 16711941 TI - Adiabatic N-soliton interactions of Bose-Einstein condensates in external potentials. AB - A perturbed version of the complex Toda chain (CTC) has been employed to describe adiabatic interactions within an N-soliton train of the nonlinear Schrodinger equation. Perturbations induced by weak quadratic and periodic external potentials are studied by both analytical and numerical means. It is found that the perturbed CTC adequately models the N-soliton train dynamics for both types of potentials. As an application of the developed theory, we consider the dynamics of a train of matter-wave solitons confined to a parabolic trap and optical lattice, as well as tilted periodic potentials. In the last case, we demonstrate that there exist critical values of the strength of the linear potential for which one or more localized states can be extracted from a soliton train. An analytical expression for these critical strengths for expulsion is also derived. PMID- 16711942 TI - Modulational instability and pattern formation in discrete dissipative systems. AB - We report in this paper the study of modulated wave trains in the one-dimensional (1D) discrete Ginzburg-Landau model. The full linear stability analysis of the nonlinear plane wave solutions is performed by considering both the wave vector (q) of the basic states and the wave vector (Q) of the perturbations as free parameters. In particular, it is shown that a threshold exists for the amplitude and above this threshold, the induced modulational instability leads to the formation of ordered and disordered patterns. The theoretical findings have been numerically tested through direct simulations and have been found to be in agreement with the theoretical prediction. We show numerically that modulational instability is also an indicator of the presence of discrete solitons as were early predicted to exist in Ginzburg-Landau lattices. PMID- 16711943 TI - Plane-wave solutions to frequency-domain and time-domain scattering from magnetodielectric slabs. AB - Plane-wave representations are used to formulate the exact solutions to frequency domain and time-domain sources illuminating a magnetodielectric slab with complex permittivity epsilon(omega) and permeability mu(omega). In the special case of a line source at z = 0 a distance d < L in front of an L-wide lossless double negative (DNG) slab with kappa(omega 0) = epsilon(omega 0) / epsilon 0 = mu(omega 0) / mu 0 = (-1), the single-frequency (omega 0) solution exhibits not only "perfectly focused" fields for z > 2L but also divergent infinite fields in the region 2d < z < 2L. In contrast, the solution to the same lossless kappa(omega 0) = (-1) DNG slab illuminated by a sinusoidal wave that begins at some initial time t = 0 (and thus has a nonzero bandwidth, unlike the single-frequency excitation that begins at t = (-infinity) is proven to have imperfectly focused fields and convergent finite fields everywhere for all finite time t. The proof hinges on the variation of kappa(omega) about omega = omega 0 having a lower bound imposed by causality and energy conservation. The minimum time found to produce a given resolution is proportional to the estimate obtained by G. Gomez-Santos, [Phys. Rev. Lett. 90, 077401 (2003)]. Only as t --> infinity do the fields become perfectly focused in the region z > 2L and divergent in the region 2d < z < 2L. These theoretical results, which are confirmed by numerical examples, imply that divergent fields of the single-frequency solution are not caused by an inherent inconsistency in assuming an ideal lossless kappa(omega 0) = (-1) DNG material, but are the result of the continuous single-frequency wave (which contains infinite energy) building up infinite reactive fields during the infinite duration of time from t = (-infinity) to the present time t that the single frequency excitation has been applied. An analogous situation occurs at the resonant frequencies of a lossless cavity. A single-frequency (zero-bandwidth) source inside the cavity produces infinite fields at a resonant frequency, whereas the same source turned on at time t = 0 (so that it has a nonzero bandwidth) produces finite fields. PMID- 16711944 TI - Scaling, self-similar solutions and shock waves for V-shaped field potentials. AB - We investigate a (1+1)-dimensional nonlinear field theoretic model with the field potential V(phi) = absolute value of phi. It can be obtained as the universal small amplitude limit in a class of models with potentials which are symmetrically V-shaped at their minima, or as a continuum limit of certain mechanical system with an infinite number of degrees of freedom. We point out that the model has an interesting scaling symmetry. One consequence of that is the existence of self-similar solutions of the corresponding field equations. We also find discontinuous solutions of shock wave type. PMID- 16711945 TI - Photonic band-gap formation by optical-phase-mask lithography. AB - We demonstrate an approach for fabricating photonic crystals with large three dimensional photonic band gaps (PBG's) using single-exposure, single-beam, optical interference lithography based on diffraction of light through an optical phase mask. The optical phase mask (OPM) consists of two orthogonally oriented binary gratings joined by a thin, solid layer of homogeneous material. Illuminating the phase mask with a normally incident beam produces a five-beam diffraction pattern which can be used to expose a suitable photoresist and produce a photonic crystal template. Optical-phase-mask Lithography (OPML) is a major simplification from the previously considered multibeam holographic lithography of photonic crystals. The diffracted five-beam intensity pattern exhibits isointensity surfaces corresponding to a diamondlike (face-centered cubic) structure, with high intensity contrast. When the isointensity surfaces in the interference patterns define a silicon-air boundary in the resulting photonic crystal, with dielectric contrast 11.9 to 1, the optimized PBG is approximately 24% of the gap center frequency. The ideal index contrast for the OPM is in the range of 1.7-2.3. Below this range, the intensity contrast of the diffraction pattern becomes too weak. Above this range, the diffraction pattern may become too sensitive to structural imperfections of the OPM. When combined with recently demonstrated polymer-to-silicon replication methods, OPML provides a highly efficient approach, of unprecedented simplicity, for the mass production of large scale three-dimensional photonic band-gap materials. PMID- 16711946 TI - Statistical analysis of noise-induced multiple filamentation. AB - The propagation of high-power large-aperture laser beams in a Kerr medium is considered. A statistical approach is developed for the growth of filaments from small-amplitude small-scale initial modulations. Closed-form expressions are derived for the intensity distribution, contrast, and maximal beam intensity, which are valid up to the blowup of the most intense filament. Numerical experiments are found to be in good agreement with theoretical predictions. PMID- 16711947 TI - Nonlocal homogenization model for a periodic array of epsilon-negative rods. AB - We propose an effective permittivity model to homogenize an array of long thin epsilon-negative rods arranged in a periodic lattice. It is proven that the effect of spatial dispersion in this electromagnetic crystal cannot be neglected, and that the medium supports dispersionless modes that guide the energy along the rod axes. It is suggested that this effect may be used to achieve subwavelength imaging at the infrared and optical domains. The reflection problem is studied in detail for the case in which the rods are parallel to the interfaces. Full wave numerical simulations demonstrate the validity and accuracy of the new model. PMID- 16711948 TI - Superluminal propagation of solitary kinklike waves in amplifying media. AB - It is shown that solitary-wave, kinklike structures can propagate superluminally in two- and four-level amplifying media with strongly damped oscillations of coherences. This is done by solving analytically the Maxwell-Bloch equations in the kinetic limit. It is also shown that the true wave fronts--unlike the pseudo wave fronts of the kinks--must propagate with velocity c, so that no violation of special relativity is possible. The conditions of experimental verification are discussed. PMID- 16711949 TI - Multidimensional reactive scattering with quantum trajectories. AB - Ensembles of quantum trajectories are evolved to study time-dependent reaction dynamics in multidimensional systems with up to 25 vibrational modes. The equations of motion are formulated in curvilinear reaction path coordinates and all coupling terms are retained, including those involving curvature of the reaction path. The model potential is a Gaussian barrier along the translational coordinate coupled to M vibrational modes. Spatial derivatives needed to propagate the trajectories are evaluated by least squares fitting in a contracted basis set. Stable propagation of the trajectory ensembles was carried out until complete bifurcation into reflected and reactive subensembles. The reaction probabilities were evaluated by Monte Carlo integration of the multidimensional smooth transmitted densities. Computational results, including trajectory plots and time-dependent reaction probabilities, are presented for M = 1, 5, and 25 vibrational modes. PMID- 16711951 TI - Linear relation on the correlation in complex networks. AB - Correlation in complex networks follows a linear relation between the degree of a node and the total degrees of its neighbors for six different classes of real networks. This general linear relation is an extension of the Aboav-Weaire law in two-dimensional cellular structures and provides a simple and different perspective on the correlation in complex networks, which is complementary to an existing description using Pearson correlation coefficients and a power law fit. Analytical expression for this linear relation for three standard models of complex networks: the Erdos-Renyi, Watts-Strogatz, and Barabasi-Albert networks is provided. The slope and intercept of this linear relation are described by a single parameter a together with the first and second moment of the degree distribution of the network. The assortivity of the network can be related to the sign of the intercept. PMID- 16711950 TI - Quantum Monte Carlo method using a stochastic Poisson solver. AB - The quantum Monte Carlo (QMC) technique is an extremely powerful method to treat many-body systems. Usually the quantum Monte Carlo method has been applied in cases where the interaction potential has a simple analytic form, like the 1/r Coulomb potential. However, in a complicated environment as in a semiconductor heterostructure, the evaluation of the interaction itself becomes a nontrivial problem. Obtaining the potential from any grid-based finite-difference method for every walker and every step is infeasible. We demonstrate an alternative approach of solving the Poisson equation by a classical Monte Carlo calculation within the overall quantum Monte Carlo scheme. We have developed a modified "walk on spheres" algorithm using Green's function techniques, which can efficiently account for the interaction energy of walker configurations, typical of quantum Monte Carlo algorithms. This stochastically obtained potential can be easily incorporated with variational, diffusion, and other Monte Carlo techniques. We demonstrate the validity of this method by studying a simple problem, the polarization of a helium atom in the electric field of an infinite capacitor. PMID- 16711952 TI - Probing criticality with linearly varying external fields: Renormalization group theory of nonequilibrium critical dynamics under driving. AB - A renormalization group theory for the nonlinear and nonequilibrium responses to linearly varying external probes is formulated for the critical dynamics of a time-dependent Ginzburg-Landau equation with a scalar nonconserved order parameter. A series of nonequilibrium dynamic critical exponents and their scaling relations characterizing the time-dependent probes and the nonequilibrium hysteresis induced are derived analytically and systematically and agree well with numerical results. The three-dimensional dynamic critical exponent is accordingly determined as z = 2.036(11). These show that the linearly varying external fields can be applied to probe criticality effectively. PMID- 16711953 TI - Phase transition of triplet reaction-diffusion models. AB - The phase transitions classes of reaction-diffusion systems with multiparticle reactions are an open challenging problem. Large scale simulations are applied for the 3A --> 4A, 3A --> 2A and the 3A --> 4A, 3A --> [formula : see text] triplet reaction models with site occupation restriction in one dimension. Static and dynamic mean-field scaling are observed with signs of logarithmic corrections suggesting d(c) = 1 upper critical dimension for this family of models. PMID- 16711954 TI - Monte Carlo results in time-dependent hierarchical fiber-bundle models of fracture. AB - Is there any time threshold of stability in a time-dependent hierarchical load transfer structure formed by N elements, in the limit N --> infinity? There is no rigorous proof, but the consensus to this question is yes. Here we extend our previous work on these systems up to a size N = 10(6), using a power law breakdown rule and a new more efficient Monte Carlo method. The new results confirm this positive consensus. PMID- 16711955 TI - Synchronization properties of two self-oscillating semiconductor lasers subject to delayed optoelectronic mutual coupling. AB - We theoretically investigate the nonlinear dynamics and synchronization properties between two mutually coupled semiconductor lasers units. Each unit can self-oscillate by means of delayed optoelectronic feedback loops. The mutual optoelectronic interactions between the laser units take into account the finite propagation time of the signals. Under perfectly symmetric conditions, we find different "death by delay" islands that persist for instantaneous coupling. The appearance of (zero lag) isochronous chaotic synchronization, under appropriate driving conditions, is another distinctive feature of the delayed feedback loops in the laser units. For slightly asymmetric operation, we obtain frequency locked bands (Arnold Tongue) whose width periodically changes with the coupling delay time. PMID- 16711956 TI - Wigner surmises and the two-dimensional homogeneous Poisson point process. AB - We derive a set of identities that relate the higher-order interpoint spacing statistics of the two-dimensional homogeneous Poisson point process to the Wigner surmises for the higher-order spacing distributions of eigenvalues from the three classical random matrix ensembles. We also report a remarkable identity that equates the second-nearest-neighbor spacing statistics of the points of the Poisson process and the nearest-neighbor spacing statistics of complex eigenvalues from Ginibre's ensemble of 2 x 2 complex non-Hermitian random matrices. PMID- 16711957 TI - Bivariate- distribution for transition matrix elements in Breit-Wigner to Gaussian domains of interacting particle systems. AB - Interacting many-particle systems with a mean-field one-body part plus a chaos generating random two-body interaction having strength lambda exhibit Poisson to Gaussian orthogonal ensemble and Breit-Wigner (BW) to Gaussian transitions in level fluctuations and strength functions with transition points marked by lambda = lambda c and lambda = lambda F, respectively; lambda F >> lambda c. For these systems a theory for the matrix elements of one-body transition operators is available, as valid in the Gaussian domain, with lambda > lambda F, in terms of orbital occupation numbers, level densities, and an integral involving a bivariate Gaussian in the initial and final energies. Here we show that, using a bivariate-t distribution, the theory extends below from the Gaussian regime to the BW regime up to lambda = lambda c. This is well tested in numerical calculations for 6 spinless fermions in 12 single-particle states. PMID- 16711958 TI - Analytical investigation of the combined effect of fluid inertia and unsteadiness on low-Re particle centrifugation. AB - We analyze the explicit contribution of fluid inertia and fluid unsteadiness to the force acting on a solid sphere moving in a vertical solid-body rotation flow, in the limit of small Reynolds and Taylor numbers. This problem can be thought of as a test case where the flow induced by the particle is both unsteady (in the laboratory frame) and convected by the unperturbed flow. Many authors assume that the contributions of these two effects can be approximately superposed, and postulate that the particle motion equation is composed of the classical Boussinesq-Basset-Oseen equation (obtained by neglecting the fluid inertia) plus an additive lift force. In the present paper the simplicity of the unperturbed flow enables one to calculate analytically the explicit contribution of each term appearing in the perturbed flow equation (by using matched asymptotic expansions). Our results show how the convective terms and the unsteady term do contribute to the particle drag and lift coefficients in a very complex and nonadditive manner. PMID- 16711959 TI - Onset of convection in Soret-driven instability. AB - It is common knowledge that light fluids rise while heavy fluids sink in the gravity field. The most obvious case is the isothermal Rayleigh-Taylor instability when a heavy fluid is placed on top of a light one. In the nonisothermal case, while heating from above, the density stratification is stable in a pure liquid. However, unstable density stratification might be established in a binary mixture with a negative Soret effect in the case of heating from above: the heavier liquid is accumulated on the top of the lighter one. Due to the large differences between viscous, thermal, and diffusion times the system has a tendency to fingering buoyant instabilities. At some moment the flow may be initiated. Near the onset of convection the flow pattern has a columnar convective structure: for a relatively low applied temperature difference Delta T the lighter and colder liquid is drawn up in the central part of the cell and the heavier liquid flows down along the walls. For finite size systems the situation is reversed at higher Delta T. Here we present results of three-dimensional direct numerical simulations of heat and mass transfer in a system with a negative Soret effect. While the development of Soret-induced convection is similar for a wide class of liquids: water based mixtures, colloidal, and polymer solutions, the parameters of the chosen system correspond to a realistic binary mixture of water (90%) and isopropanol (10%) enabling comparison of theoretical predictions with planned experimental studies. PMID- 16711960 TI - Size distribution and energy spectrum in the mixed state induced by Rayleigh Taylor instability. AB - A study--based on simulations and experiments as well as analytical derivations- of the internal structure of the fragmented ("mixed") state induced by the Rayleigh-Taylor instability at the interface between two fluids is presented. The distribution of sizes and the energy spectrum in the fragmented state are derived from the symmetries exhibited by the data and by dimensional analysis. PMID- 16711961 TI - Hugoniot data of plastic foams obtained from laser-driven shocks. AB - In this paper we present Hugoniot data for plastic foams obtained with laser driven shocks. Relative equation-of-state data for foams were obtained using Al as a reference material. The diagnostics consisted in the detection of shock breakout from double layer Al/foam targets. The foams [poly(4-methyl-1-pentene) with density 130 > rho > 60 mg/cm3] were produced at the Institute of Laser Engineering of Osaka University. The experiment was performed using the Prague PALS iodine laser working at 0.44 microm wavelength and irradiances up to a few 10(14) W/cm2. Pressures as high as 3.6 Mbar (previously unreached for such low density materials) where generated in the foams. Samples with four different values of initial density were used, in order to explore a wider region of the phase diagram. Shock acceleration when the shock crosses the Al/foam interface was also measured. PMID- 16711962 TI - Structure of multispecies charged particles in a quadratic trap. AB - Multispecies interacting charged particles in a two-dimensional quadratic trap are studied. The ground-state configurations for different particle and species numbers are obtained by molecular dynamics simulation. It is found that particles with similar mass-to-charge ratio tend to populate a common shell, whose location depends on the particle mass-to-charge ratio, and that the greater the latter, the closer are the particles to the center of the trap. This scaling for the ground-state configuration is independent of the total particle and species numbers in the system. PMID- 16711963 TI - Analysis and reduction of the spurious current in a class of multiphase lattice Boltzmann models. AB - We show that the spurious current present near a curved interface in a class of multiphase lattice Boltzmann (LB) models is due to the insufficient isotropy of the discrete gradient operator. A method of obtaining highly isotropic gradient operators on a lattice is given. Numerical simulations show that both the magnitude and the spatial extent of the spurious current are significantly reduced as gradient operators of increasingly higher order of isotropy is adopted in multiphase LB models. PMID- 16711964 TI - Pressure boundary condition of the lattice Boltzmann method for fully developed periodic flows. AB - The general periodic boundary condition for the lattice Boltzmann method has been modified to incorporate the pressure difference for fully developed periodic flows. The results demonstrated that, unlike other existing pressure boundary treatments, the proposed procedure/treatment does not generate nonphysical inlet and outlet flow disturbances while preserving the system periodicity. This method is readily applicable to a range of lattice Boltzmann simulations for systems with periodic electric potential and temperature fields. PMID- 16711965 TI - Comment on "Low-frequency character of the Casimir force between metallic films". AB - In Phys. Rev. E 70, 047102 (2004), Torgerson and Lamoreaux investigated for the first time the real-frequency spectrum of the finite temperature correction to the Casimir force, for metallic plates of finite conductivity. The very interesting result of this study is that the large correction from the TE mode is dominated by low frequencies, for which the dielectric description of the metal is invalid, and the authors correctly point out that a more realistic description is provided by low-frequency metallic boundary conditions. However, their subsequent analysis uses an incorrect form of metallic boundary conditions for TE modes. After correcting this error, we find that their main conclusion was nevertheless qualitatively correct: contrary to the result of the dielectric model, the thermal TE mode correction leads to an increase in the TE mode force of attraction between the plates. The correction found by us, however, has a magnitude about 20 times larger than that quoted by Torgerson and Lamoreaux. PMID- 16711967 TI - Comment on "Structure of positive decompositions of exponential operators". AB - An elementary proof is shown on the necessary existence of negative coefficients in splitting methods of order p > or = 3. PMID- 16711968 TI - Pfaffian pairing wave functions in electronic-structure quantum Monte Carlo simulations. AB - We investigate the accuracy of trial wave functions for quantum Monte Carlo based on Pfaffian functional form with singlet and triplet pairing. Using a set of first row atoms and molecules we find that these wave functions provide very consistent and systematic behavior in recovering the correlation energies on the level of 95%. In order to get beyond this limit we explore the possibilities of multi-Pfaffian pairing wave functions. We show that a small number of Pfaffians recovers another large fraction of the missing correlation energy comparable to the larger-scale configuration interaction wave functions. We also find that Pfaffians lead to substantial improvements in fermion nodes when compared to Hartree-Fock wave functions. PMID- 16711969 TI - Density profile of a trapped strongly interacting Fermi gas with unbalanced spin populations. AB - We present a theoretical study of the density profile of a trapped strongly interacting Fermi gas with unbalanced spin populations. Making the assumption of the existence of a first order phase transition between an unpolarized superfluid phase and a fully polarized normal phase, we show good agreement with a recent experiment presented by Partridge et al. PMID- 16711970 TI - Momentum transfer from quantum vacuum to magnetoelectric matter. AB - A recent publication [Phys. Rev. Lett. 92, 020404 (2004)] raises the possibility of momentum transfer from zero-point quantum fluctuations to matter, controlled by applied electric and magnetic fields. We present a Lorentz-invariant description using field-theoretical regularization techniques. We find no momentum transfer for homogeneous media, but predict a very small transfer for a Casimir-type geometry. PMID- 16711971 TI - Observations of density fluctuations in an elongated Bose gas: ideal gas and quasicondensate regimes. AB - We report in situ measurements of density fluctuations in a quasi-one-dimensional 87Rb Bose gas at thermal equilibrium in an elongated harmonic trap. We observe an excess of fluctuations compared to the shot-noise level expected for uncorrelated atoms. At low atomic density, the measured excess is in good agreement with the expected "bunching" for an ideal Bose gas. At high density, the measured fluctuations are strongly reduced compared to the ideal gas case. We attribute this reduction to repulsive interatomic interactions. The data are compared with a calculation for an interacting Bose gas in the quasicondensate regime. PMID- 16711972 TI - Noise thermometry with two weakly coupled Bose-Einstein condensates. AB - Here we report on the experimental investigation of thermally induced fluctuations of the relative phase between two Bose-Einstein condensates which are coupled via tunneling. The experimental control over the coupling strength and the temperature of the thermal background allows for the quantitative analysis of the phase fluctuations. Furthermore, we demonstrate the application of these measurements for thermometry in a regime where standard methods fail. With this we confirm that the heat capacity of an ideal Bose gas deviates from that of a classical gas as predicted by the third law of thermodynamics. PMID- 16711973 TI - Entangled photon pairs from semiconductor quantum dots. AB - Tomographic analysis demonstrates that the polarization state of pairs of photons emitted from a biexciton decay cascade becomes entangled when spectral filtering is applied. The measured density matrix of the photon pair satisfies the Peres criterion for entanglement by more than 3 standard deviations of the experimental uncertainty and violates Bell's inequality. We show that the spectral filtering erases the "which path" information contained in the photons' color and that the remanent information in the quantum dot degrees of freedom is negligible. PMID- 16711974 TI - Quantum teleportation with a three-Bell-state analyzer. AB - We present a novel Bell-state analyzer (BSA) for time-bin qubits allowing the detection of three out of four Bell states with linear optics, two detectors, and no auxiliary photons. The theoretical success rate of this scheme is 50%. Our new BSA demonstrates the power of generalized quantum measurements, known as positive operator valued measurements. A teleportation experiment was performed to demonstrate its functionality. We also present a teleportation experiment with a fidelity larger than the cloning limit. PMID- 16711975 TI - Signature of directed chaos in the conductance of a nanowire. AB - We study the conductance of chaotic or disordered wires in a situation where equilibrium transport decomposes into biased diffusion and a countermoving regular current. A possible realization is a semiconductor nanostructure with a transversal magnetic field and suitably patterned surfaces. We find a nontrivial dependence of the conductance on the wire length. It differs qualitatively from Ohm's law by the existence of a characteristic length scale and a finite saturation value. PMID- 16711976 TI - Adiabatic swimming in an ideal quantum gas. AB - Interference effects are important for swimming of mesoscopic systems that are small relative to the coherence length of the surrounding quantum medium. Swimming is geometric for slow swimmers and the distance covered in each stroke is determined, explicitly, in terms of the on-shell scattering matrix. Remarkably, for a one-dimensional Fermi gas at zero temperature we find that slow swimming is topological: the swimming distance covered in one stroke is quantized in half integer multiples of the Fermi wavelength. In addition, a careful choice of the swimming stroke can eliminate dissipation. PMID- 16711977 TI - Entropic transport: kinetics, scaling, and control mechanisms. AB - We show that transport in the presence of entropic barriers exhibits peculiar characteristics which makes it distinctly different from that occurring through energy barriers. The constrained dynamics yields a scaling regime for the particle current and the diffusion coefficient in terms of the ratio between the work done to the particles and available thermal energy. This interesting property, genuine to the entropic nature of the barriers, can be utilized to effectively control transport through quasi-one-dimensional structures in which irregularities or tortuosity of the boundaries cause entropic effects. The accuracy of the kinetic description has been corroborated by simulations. Applications to different dynamic situations involving entropic barriers are outlined. PMID- 16711978 TI - Thermal spectral functions of strongly coupled N = 4 supersymmetric Yang-Mills theory. AB - We use the gauge-gravity duality conjecture to compute spectral functions of the stress-energy tensor in finite-temperature N = 4 supersymmetric Yang-Mills theory in the limit of large N(c) and large 't Hooft coupling. The spectral functions exhibit peaks characteristic of hydrodynamic modes at small frequency, and oscillations at intermediate frequency. The nonperturbative spectral functions differ qualitatively from those obtained in perturbation theory. The results may prove useful for lattice studies of transport processes in thermal gauge theories. PMID- 16711979 TI - Mass and width of the lowest resonance in QCD. AB - We demonstrate that near the threshold, the pi pi scattering amplitude contains a pole with the quantum numbers of the vacuum--commonly referred to as the sigma- and determine its mass and width within small uncertainties. Our derivation does not involve models or parametrizations but relies on a straightforward calculation based on the Roy equation for the isoscalar S wave. PMID- 16711980 TI - Isolated photons in deep inelastic scattering. AB - Photon radiation at large transverse momenta at colliders is a detailed probe of hard interaction dynamics. The isolated photon production cross section in deep inelastic scattering was measured recently by the ZEUS experiment, and found to be considerably larger than theoretical predictions obtained with widely used event generators. To investigate this discrepancy, we perform a dedicated parton level calculation of this observable, including contributions from fragmentation and large-angle radiation. Our results are in good agreement with all aspects of the experimental measurement. PMID- 16711981 TI - Evidence for nucleon-resonance excitation in omega-meson photoproduction. AB - The photoproduction of the omega meson has been studied at GRAAL from threshold up to a photon energy of 1.5 GeV. The differential cross sections and beam asymmetries have been measured precisely at all angles. The total cross section is also obtained. Systematic enhancements of the differential cross section at large angles and nonzero beam asymmetries at intermediate angles provide clear evidence for s- and u-channel resonant processes. The data are compared to the results of hadron and quark models. PMID- 16711982 TI - Matter formed at the BNL Relativistic Heavy Ion Collider. AB - We suggest that the "new form of matter" found just above T(c) by the Relativistic Heavy Ion Collider is made up of tightly bound quark-antiquark pairs, essentially 32 chirally restored (more precisely, nearly massless) mesons of the quantum numbers of pi, sigma, rho, and a1. Taking the results of lattice gauge simulations (LGS) for the color Coulomb potential from the work of the Bielefeld group and feeding this into a relativistic two-body code, after modifying the heavy-quark lattice results so as to include the velocity-velocity interaction, all ground-state eigenvalues of the 32 mesons go to zero at T(c) just as they do from below T(c) as predicted by the vector manifestation of hidden local symmetry. This could explain the rapid rise in entropy up to T(c) found in LGS calculations. We argue that how the dynamics work can be understood from the behavior of the hard and soft glue. PMID- 16711983 TI - Direct empirical correlation between proton-neutron interaction strengths and the growth of collectivity in nuclei. AB - A direct correlation between experimental values of proton-neutron interaction strengths and experimental measures of the growth of collectivity in nuclei is found. In particular, differences in the p-n interaction strengths and differences in growth rates of collectivity in particle-particle (or hole-hole) and particle-hole regions are found to correspond. PMID- 16711984 TI - High-energy cutoff in the spectrum of strong-field nonsequential double ionization. AB - Electron energy distributions of singly and doubly ionized helium in an intense 390 nm laser field have been measured at two intensities (0.8 PW/cm2 and 1.1 PW/cm2, where PW is defined as 10(15) W/cm2). Numerical solutions of the full dimensional time-dependent helium Schrodinger equation show excellent agreement with the experimental measurements. The high-energy portion of the two-electron energy distributions reveals an unexpected 5U(p) cutoff for the double ionization (DI) process and leads to a proposed model for DI below the quasiclassical threshold. PMID- 16711985 TI - Generating controllable atom-light entanglement with a Raman atom laser system. AB - We introduce a scheme for creating continuous variable entanglement between an atomic beam and an optical field, by using squeezed light to outcouple atoms from a Bose-Einstein condensate via a Raman transition. We model the full multimode dynamics of the atom laser beam and the squeezed optical field and show that, with appropriate two-photon detuning and two-photon Rabi frequency, the transmitted light is entangled in amplitude and phase with the outcoupled atom laser beam. The degree of entanglement is controllable via changes in the two photon Rabi frequency of the outcoupling process. PMID- 16711986 TI - Quantum noise minimization in transistor amplifiers. AB - General quantum restrictions on the noise performance of linear transistor amplifiers are used to identify the region in parameter space where the quantum limited performance is achievable and to construct a practical procedure for approaching it experimentally using only the knowledge of directly measurable quantities: the gain, (differential) conductance, and the output noise. A specific example of resonant barrier transistors is discussed. PMID- 16711987 TI - Collapse of optical vortices. AB - We theoretically and experimentally investigate the self-focusing of optical vortices in Kerr media. We observe collapse to a distinct self-similar profile, which becomes unstable to azimuthal perturbations. We analyze the azimuthal modulational instability for ring-shaped vortices and predict the number of azimuthal maxima solely as a function of power and topological charge. In our experiments, the observed multiple-filamentation patterns are in excellent agreement with our theoretical analysis. PMID- 16711988 TI - Fast relaxation of carbon nanotubes in polymer composite actuators. AB - Silicone elastomer composites containing multiwalled carbon nanotubes have been irradiated with near-infrared light to study their mechanical actuation response. We show that the speed of the stimulated response is faster than Debye relaxation, instead following a compressed-exponential law. However, the relaxation after switching off the light source follows the simple-exponential relaxation, as does the stimulated response at very low nanotube concentration. We discuss possible models and explanations to account for the fast photomechanical response. PMID- 16711989 TI - Deterministic ratchets, circle maps, and current reversals. AB - In this work we transform the deterministic dynamics of an overdamped tilting ratchet into a discrete dynamical map by looking stroboscopically at the continuous motion originally ruled by differential equations. We show that, for the simple and widely used case of periodic dichotomous driving forces, the resulting discrete map belongs to the class of circle homeomorphisms. This approach allows us to apply the well-known properties of such maps to derive the necessary and sufficient conditions that the ratchet potential must satisfy in order to have a vanishing current. Furthermore, as a consequence of the above, we show (i) that there is a class of periodic potentials which do not exhibit the rectification phenomenon in spite of their asymmetry and (ii) that current reversals occur in the deterministic case for a large class of ratchet potentials. PMID- 16711990 TI - Dust eruptions by photophoresis and solid state greenhouse effects. AB - We carried out experiments that show a gas pressure dependent ability of light to eject particles from a dust bed. Dust eruptions also occur upon removal of the light source. This can be attributed to a solid state greenhouse effect and photophoretic forces. This ejection mechanism works at light intensities larger than 6 kW/m2 but in extreme cases might work as low as 1 kW/m2. It can be applied to sunlit dust on Mars where it aids or triggers dust lift-off from the surface into the atmosphere. It is of importance for dusty bodies at the inner edge of protoplanetary disks where it leads to light induced erosion. The effect also offers a base for technical applications of dust removal in low pressure environments. PMID- 16711991 TI - Resolving singular forces in cavity flow: multiscale modeling from atomic to millimeter scales. AB - Flow driven by moving a wall that bounds a fluid-filled cavity is a classic example of a multiscale problem. Continuum equations predict that every scale contributes roughly equally to the total force on the moving wall, leading to a logarithmic divergence, and that there is an infinite hierarchy of vortices at the stationary corners. A multiscale approach is developed that retains an atomistic description in key regions. Following the stress over more than six decades in length in systems with characteristic scales of up to millimeters and milliseconds allows us to resolve the singularities and determine the force for the first time. We find a universal dependence on the macroscopic Reynolds number, and large atomistic effects that depend on wall velocity and interactions. PMID- 16711992 TI - Multipoint holographic optical velocimetry in microfluidic systems. AB - We show how holographic optical trapping can be used for the multipoint measurement of fluid flow in microscopic geometries. An array of microprobes can be simultaneously trapped and used to map out the fluid flow in a microfluidic device. The optical traps are alternately turned on and off such that the probe particles are displaced by the flow of the surrounding fluid and then retrapped. The particles' displacements are monitored by digital video microscopy and directly converted into velocity field values. This technique enables the measurement of a two-dimensional flow field at points arbitrarily distributed in a three-dimensional volume. The validity of the technique is demonstrated for the case of the flow around a spinning sphere and the flow at the outlet of a microchannel. PMID- 16711993 TI - Centrifugal forces alter streamline topology and greatly enhance the rate of heat and mass transfer from neutrally buoyant particles to a shear flow. AB - Centrifugal forces break the degenerate closed-streamline configuration that occurs in simple shear flow past a neutrally buoyant torque-free particle in the inertialess limit. The broken symmetry allows heat or mass to be convected away in an efficient manner in sharp contrast to the inertialess diffusion-limited scenario. The dimensionless transfer rate, characterized by the Nusselt number, is found to be Nu = 0.33(RePe)(1/3) + O(1) for small but finite Re when RePe >> 1. Here, the particle Reynolds number (Re) is a dimensionless measure of the inertial forces, while the Peclet number (Pe) measures the relative importance of the convective and the diffusive transfer mechanisms. The symmetry-breaking bifurcation is expected to occur in generic shearing flows, and represents a possible means for heat or mass transfer enhancement from the dispersed phase in multiphase systems. PMID- 16711994 TI - Rayleigh-Taylor turbulence in two dimensions. AB - The first consistent phenomenological theory for two- and three-dimensional Rayleigh-Taylor (RT) turbulence has recently been presented by Chertkov [Phys. Rev. Lett. 91, 115001 (2003)]. By means of direct numerical simulations, we confirm the spatiotemporal prediction of the theory in two dimensions and explore the breakdown of the phenomenological description due to intermittency effects. We show that small-scale statistics of velocity and temperature follow Bolgiano Obukhov scaling. At the level of global observables, we show that the time dependent Nusselt and Reynolds numbers scale as the square root of the Rayleigh number. These results point to the conclusion that RT turbulence in two and three dimensions, thanks to the absence of boundaries, provides a natural physical realization of the Kraichnan scaling regime hitherto associated with the elusive "ultimate state of thermal convection." PMID- 16711995 TI - Selection of two-phase flow patterns at a simple junction in microfluidic devices. AB - We study the behavior of a confined stream made of two immiscible fluids when it reaches a T junction. Two flow patterns are witnessed: the stream is either directed in only one sidearm, yielding a preferential flow pathway for the dispersed phase, or splits between both. We show that the selection of these patterns is not triggered by the shape of the junction nor by capillary effects, but results from confinement. It can be anticipated in terms of the hydrodynamic properties of the flow. A simple model yielding universal behavior in terms of the relevant adimensional parameters of the problem is presented and discussed. PMID- 16711996 TI - Quasiadiabatic, nonfocusing transition-energy crossing. AB - A quasiadiabatic nonfocusing transition-energy crossing is proposed for suppressing any nonadiabatic and undesired features in a longitudinally separated function-type accelerator, in which particles are confined by an radio-frequency voltage with an adiabatic reduction of the amplitude and accelerated by a step voltage. This new method has been examined, both theoretically and experimentally. PMID- 16711997 TI - Coalescence of magnetic islands in the low-resistivity, Hall-MHD regime. AB - The coalescence of magnetic islands in the low-resistivity eta, Hall-MHD regime is studied. The interaction between the ion inertial length d(i) and the dynamically evolving current sheet scale length deltaJ is established. Initially, d(i) << deltaJ. If eta is such that deltaJ dynamically thins down to d(i) prior to the well-known sloshing phenomena, then sloshing is avoided. This results in eta independent peak reconnection rates. However, if d(i) is small enough that deltaJ cannot be thinned down to this scale prior to sloshing, then sloshing proceeds as in the resistive MHD model. PMID- 16711998 TI - Second-sound spectroscopy of a nonequilibrium superfluid-normal interface. AB - An experiment is proposed to test a previously developed theory of the hydrodynamics of a nonequilibrium heat current-induced superfluid-normal interface. It is shown that the interfacial "trapped" second-sound mode predicted by the theory leads to a sharp resonant dip in the reflected signal from an external second-sound pulse propagated toward the interface when its horizontal phase speed matches that of the interface mode. The influence of the interface on thermal fluctuations in the bulk superfluid is shown to lead to slow power dependence of the order parameter, and other quantities, on distance from it. PMID- 16711999 TI - High-frequency dynamics in metallic glasses. AB - Using inelastic x-ray scattering we studied the collective dynamics of the glassy alloy Ni33Zr67 in the first pseudo-Brillouin-zone, an energy-momentum region still unexplored in metallic glasses. We determine key properties such as the momentum transfer dependence of the sound velocity and of the acoustic damping, discussing the results in the general context of recently proposed pictures for acoustic dynamics in glasses. Specifically, we demonstrate the existence in this strong glass of well defined (in the Ioffe-Regel sense) acoustic-like excitations well above the boson peak energy. PMID- 16712000 TI - Do current-density nonlinearities cut off the glass transition? AB - Extended mode-coupling theories for dense fluids predict that nonlinear current density couplings cut off the singular "ideal glass transition" present in the standard mode-coupling theory where such couplings are ignored. We suggest here that, rather than allowing for activated processes as sometimes supposed, contributions from current-density couplings are always negligible close to a glass transition. We discuss in schematic terms how activated processes can nonetheless cut off the transition by causing the memory function to become linear in correlators at late times. PMID- 16712001 TI - Size-dependent pressure-induced amorphization in nanoscale TiO2. AB - We investigated the size-dependent high-pressure phase transition behavior of nanocrystalline anatase TiO2 with synchrotron x-ray diffraction and Raman spectroscopy to 45 GPa at ambient temperature. Pressure-induced amorphization results in a high-density amorphous (HDA) form when the starting crystallite size is < 10 mm. The HDA-TiO2 transforms to a low-density amorphous form at lower pressures. Harnessing the nanometer length scale thus provides a new window for experimental investigation of amorphization in poor glass formers and a synthesis route for new amorphous materials. PMID- 16712002 TI - "Magic melters" have geometrical origin. AB - Recent experimental reports bring out extreme size sensitivity in the heat capacities of gallium and aluminum clusters. In the present work we report results of our extensive ab initio molecular dynamical simulations on Ga30 and Ga31, the pair which has shown rather dramatic size sensitivity. We trace the origin of this size sensitive heat capacities to the relative order in their respective ground state geometries. Such an effect of nature of the ground state on the characteristics of heat capacity is also seen in case of small gallium and sodium clusters, indicating that the observed size sensitivity is a generic feature of small clusters. PMID- 16712003 TI - Local electromigration model for crystal surfaces. AB - The dynamics of crystal surfaces in the presence of electromigration is analyzed. From a phase field model with a migration force which depends on the local geometry, a step model with additional contributions is derived in the kinetic boundary conditions. These contributions trigger various surface instabilities, such as step meandering, bunching, and pairing on vicinal surfaces. Experiments are discussed. PMID- 16712004 TI - Soft phonon, dynamical fluctuations, and a reversible phase transition: indium chains on silicon. AB - The In/Si(111)-(4 x 1) surface is a paradigmatic example of a quasi-one dimensional system showing a reversible structural and electronic (metal insulator) phase transition when the temperature is lowered. In this work, we use first-principles simulation techniques to uncover the atomic and electronic origin of this controversial transition. Our calculations show that the ground state consists of insulating (4 x 2) indium chains with a weak interchain coupling that induces opposite shear distortions in alternate chains. First principles molecular dynamics simulations show that the (4 x 1) <--> (8 x 2) phase transition is due to the "dynamical fluctuations" the system undergoes when, at high temperature, it fluctuates chaotically between degenerate ground states. The metallicity of the In/Si(111)-(4 x 1) surface is related to the low energy cost for the shear distortion. PMID- 16712006 TI - Statistics of crumpled paper. AB - A statistical study of crumpled paper is allowed by a minimal 1D model: a self avoiding line bent at sharp angles--in which the elastic energy resides--put in a confining potential. Many independent equilibrium configurations are generated numerically and their properties are investigated. At small confinement, the distribution of segment lengths is log-normal in agreement with previous predictions and experiments. At high confinement, the system approaches a jammed state with a critical behavior, whereas the length distribution follows a gamma law in which the parameter is predicted as a function of the number of layers in the system. PMID- 16712005 TI - Nonlinear evolution of surface morphology in InAs/AlAs superlattices via surface diffusion. AB - Continuum simulations of self-organized lateral compositional modulation growth in InAs/AlAs short-period superlattices on InP substrate are presented. The results of the simulations correspond quantitatively to the results of synchrotron x-ray diffraction experiments. The time evolution of the compositional modulation during epitaxial growth can be explained only including a nonlinear dependence of the elastic energy of the growing epitaxial layer on its thickness. From the fit of the experimental data to the growth simulations we have determined the parameters of this nonlinear dependence. It was found that the modulation amplitude does not depend on the values of the surface diffusion constants of particular elements. PMID- 16712007 TI - One-dimensional surface reconstruction as an atomic-scale template for the growth of periodically striped Ag films. AB - The role of the In/Si(111)-(4 x 1)-In surface as an atomic-scale geometrical template for the growth of Ag thin films is clarified by scanning tunneling microscopy and low energy electron diffraction. Low-temperature grown Ag films are found to have stripe structures with a transverse periodicity equal to that of indium chains of the In/Si(111)-(4 x 1)-In. The stripes exhibit a structural transformation at the thickness of 6 monolayers (ML); this relaxation allows the stripes to persist up to a thickness as large as 30 ML (approximately = 7 nm) while maintaining their mean periodicity. We attribute this stability to a coincidental matching of the periodicity and the corrugation amplitude between the Ag film and the substrate, which is realized by periodic insertion of stacking faults into a Ag fcc crystal. PMID- 16712008 TI - Enhancement of electron spin coherence by optical preparation of nuclear spins. AB - We study a large ensemble of nuclear spins interacting with a single electron spin in a quantum dot under optical excitation and photon detection. At the two photon resonance between the two electron-spin states, the detection of light scattering from the intermediate exciton state acts as a weak quantum measurement of the effective magnetic (Overhauser) field due to the nuclear spins. In a coherent population trapping state without light scattering, the nuclear state is projected into an eigenstate of the Overhauser field operator, and electron decoherence due to nuclear spins is suppressed: We show that this limit can be approached by adapting the driving frequencies when a photon is detected. We use a Lindblad equation to describe the driven system under photon emission and detection. Numerically, we find an increase of the electron coherence time from 5 to 500 ns after a preparation time of 10 micros. PMID- 16712009 TI - Metamagnetic quantum criticality in Sr3Ru2O7 studied by thermal expansion. AB - We report low-temperature thermal expansion measurements on the bilayer ruthenate Sr3Ru2O7 as a function of magnetic field applied perpendicular to the ruthenium oxide planes. The field dependence of the c-axis expansion coefficient indicates the accumulation of entropy close to 8 T, related to an underlying quantum critical point. The latter is masked by two first-order metamagnetic transitions which bound a regime of enhanced entropy. Outside this region the singular thermal expansion behavior is compatible with the predictions of the itinerant theory for a two-dimensional metamagnetic quantum critical end point. PMID- 16712010 TI - Irreversible dynamics of the phase boundary in U(Ru0.96Rh0.04)2Si2 and implications for ordering. AB - We report measurements and analysis of the specific heat and magnetocaloric effect-induced temperature changes at the phase boundary into the single magnetic field-induced phase (phase II) of U(Ru0.96Rh0.04)2Si2, which yield irreversible properties similar to those at the valence transition of Yb(1-x)Y(x)InCu4. To explain these similarities, we propose a bootstrap mechanism by which lattice parameter changes caused by an electric quadrupolar order parameter within phase II become coupled to the 5f-electron hybridization, giving rise to a valence change at the transition. PMID- 16712011 TI - First-principles description of correlation effects in layered materials. AB - We present a first-principles description of anisotropic materials characterized by having both weak (dispersionlike) and strong covalent bonds, based on the adiabatic-connection fluctuation-dissipation theorem with density functional theory. For hexagonal boron nitride the in-plane and out-of-plane bonding as well as vibrational dynamics are well described both at equilibrium and when the layers are pulled apart. Bonding in covalent and ionic solids is also described. The formalism allows us to ping down the deficiencies of common exchange correlation functionals and provides insight toward the inclusion of dispersion interactions into the correlation functional. PMID- 16712012 TI - Validity of the Franck-Condon principle in the optical spectroscopy: optical conductivity of the Frohlich polaron. AB - The optical absorption of the Frohlich polaron model is obtained by an approximation-free diagrammatic Monte Carlo method and compared with two new approximate approaches that treat lattice relaxation effects in different ways. We show that: (i) a strong coupling expansion, based on the Franck-Condon principle, well describes the optical conductivity for large coupling strengths (alpha > 10); (ii) a memory function formalism with phonon broadened levels reproduces the optical response for weak coupling strengths (alpha < 6) taking the dynamic lattice relaxation into account. In the coupling regime 6 < alpha < 10, the optical conductivity is a rapidly changing superposition of both Franck Condon and dynamic contributions. PMID- 16712013 TI - Spin injection in mesoscopic silver wires: experimental test of resistance mismatch. AB - The spin polarization of current injected from a Permalloy electrode into a mesoscopic Ag wire is measured for samples with very low interface resistance. The observed value of 22.3% +/- 1.6% at 79 K is an order of magnitude larger than values previously reported for low resistance interfaces and about 4 times larger than predictions of the common resistance mismatch model. These results demonstrate that high resistance barriers are not necessary for efficient spin injection. PMID- 16712014 TI - Current-voltage characteristics of charge-ordered organic crystals. AB - The current-voltage characteristics of layered organic crystals theta-(BEDT TTF)2MZn(SCN)4 (M = Cs, Rb) follow the power law with a large exponent (e.g., 8.4 at 0.29 K for M = Cs) over a wide range of currents in the low-temperature insulating state. The power-law characteristics are attributed to electric field induced unbinding of electron-hole pairs that are thermally excited in the background of the two-dimensional charge order. The magnitude of crossover electric fields from Ohmic to the power-law characteristics indicates that the electron-electron Coulomb interaction is significantly long-ranged: The screening length is greater than 10 molecule sites. PMID- 16712015 TI - Time dependence of correlation functions following a quantum quench. AB - We show that the time dependence of correlation functions in an extended quantum system in d dimensions, which is prepared in the ground state of some Hamiltonian and then evolves without dissipation according to some other Hamiltonian, may be extracted using methods of boundary critical phenomena in d + 1 dimensions. For d = 1 particularly powerful results are available using conformal field theory. These are checked against those available from solvable models. They may be explained in terms of a picture, valid more generally, whereby quasiparticles, entangled over regions of the order of the correlation length in the initial state, then propagate classically through the system. PMID- 16712016 TI - Resonant tunneling in the fractional quantum Hall effect: superperiods and braiding statistics. AB - We study theoretically resonant tunneling of composite fermions through their quasibound states around a fractional quantum Hall island, and find a rich set of possible transitions of the island state as a function of the magnetic field or the backgate voltage. These have possible relevance to a recent experimental study, and reveal many subtleties involved in deducing fractional braiding statistics. PMID- 16712017 TI - Exciton formation and annihilation during 1D impact excitation of carbon nanotubes. AB - Near-infrared electroluminescence was recorded from unipolar single-wall carbon nanotube field-effect transistors at high drain-source voltages. High resolution spectra reveal resonant light emission originating from the radiative relaxation of excitons rather than heat dissipation. The electroluminescence is induced by only one carrier type and ascribed to 1D impact excitation. An emission quenching is also observed at high field and attributed to an exciton-exciton annihilation process and free carrier generation. The excitons' binding energy in the order of 270 meV for 1.4 nm SWNTs is inferred from the spectral features. PMID- 16712018 TI - Emission and absorption asymmetry in the quantum noise of a Josephson junction. AB - We measure current fluctuations of mesoscopic devices in the quantum regime, when the frequency is of the order of or higher than the applied voltage or temperature. Detection is designed to probe separately the absorption and emission contributions of current fluctuations, i.e. the positive and negative frequencies of the Fourier transformed nonsymmetrized noise correlator. It relies on measuring the quasiparticles photon assisted tunneling current across a superconductor-insulator-superconductor junction (the detector junction) caused by the excess current fluctuations generated by quasiparticles tunneling across a Josephson junction (the source junction). We demonstrate unambiguously that the negative and positive frequency parts of the nonsymmetrized noise correlator are separately detected and that the excess current fluctuations of a voltage biased Josephson junction present a strong asymmetry between emission and absorption. PMID- 16712019 TI - Crossover from conserving to lossy transport in circular random-matrix ensembles. AB - In a quantum dot with three leads, the transmission matrix t12 between two of these leads is a truncation of a unitary scattering matrix S, which we treat as random. As the number of channels in the third lead is increased, the constraints from the symmetry of S become less stringent and t12 becomes closer to a matrix of complex Gaussian random numbers with no constraints. We consider the distribution of the singular values of t12, which is related to a number of physical quantities. PMID- 16712021 TI - Surface-acoustic-wave-induced transport in a double quantum dot. AB - We report on nonadiabatic transport through a double quantum dot under irradiation of surface acoustic waves generated on chip. At low excitation powers, absorption and emission of single and multiple phonons are observed. At higher power, sequential phonon assisted tunneling processes excite the double dot in a highly nonequilibrium state. The present system is attractive for studying electron-phonon interaction with piezoelectric coupling. PMID- 16712020 TI - Landau-level splitting in graphene in high magnetic fields. AB - The quantum Hall (QH) effect in two-dimensional electrons and holes in high quality graphene samples is studied in strong magnetic fields up to 45 T. QH plateaus at filling factors nu = 0, +/-1, +/-4 are discovered at magnetic fields B > 20 T, indicating the lifting of the fourfold degeneracy of the previously observed QH states at nu = +/-4(absolute value(n) + 1/2), where n is the Landau level index. In particular, the presence of the nu = 0, +/-1 QH plateaus indicates that the Landau level at the charge neutral Dirac point splits into four sublevels, lifting sublattice and spin degeneracy. The QH effect at nu = +/ 4 is investigated in a tilted magnetic field and can be attributed to lifting of the spin degeneracy of the n = 1 Landau level. PMID- 16712022 TI - Temperature square dependence of the low frequency charge noise in the Josephson junction qubits. AB - To verify the hypothesis about the common origin of the low-frequency 1/f noise and the quantum f noise recently measured in the Josephson charge qubits, we study the temperature dependence of the 1/f noise and decay of coherent oscillations. The T2 dependence of the 1/f noise is experimentally demonstrated, which supports the hypothesis. We also show that dephasing in the Josephson charge qubits off the electrostatic energy degeneracy point is consistently explained by the same low-frequency 1/f noise that is observed in the transport measurements. PMID- 16712023 TI - Magnetic field effect on the superconducting magnetic gap of Nd1.85Ce0.15CuO4. AB - Inelastic neutron-scattering measurements on the archetypical electron-doped material Nd1.85Ce0.15CuO4 up to a high relative magnetic-field strength, H/H(c2) approximately 50%, reveal a simple linear magnetic-field effect on the superconducting magnetic gap and the absence of field-induced in-gap states. The extrapolated gap-closing field value is consistent with the upper critical field H(c2), and the high-field response resembles that of the paramagnetic normal state. PMID- 16712024 TI - Phason and amplitudon in the charge-density-wave phase of one-dimensional charge stripes in La(2-x)Sr(x)CuO4. AB - The present systematic Raman scattering experiments reveal the phason and amplitudon of the charge density wave (CDW) mode in the charge stripes of La(2 x)Sr(x)CuO4. Only about 15% of the electronic density of states condenses into the CDW state. The symmetries of the CDW modes change by the rotation of the stripes at the insulator-metal transition. The energy of the phason is finite at 0.06 < or = x < or = 0.1 and zero at x = 0.035 and 0.115 < or = x < or = 0.135, which suggests that the CDW is commensurate at 0.06 < or = x < or = 0.1 and incommensurate otherwise. The zero-energy phason seems to reduce T(c) at x = 1/8. PMID- 16712025 TI - Anomalous spontaneous reversal in magnetic heterostructures. AB - We observe a thermally induced spontaneous magnetization reversal of epitaxial ferromagnet/antiferromagnet heterostructures under a constant applied magnetic field. Unlike any other magnetic system, the magnetization spontaneously reverses, aligning antiparallel to an applied field with decreasing temperature. We show that this unusual phenomenon is caused by the interfacial antiferromagnetic coupling overcoming the Zeeman energy of the ferromagnet. A significant temperature hysteresis exists, whose height and width can be tuned by the field applied during thermal cycling. The hysteresis originates from the intrinsic magnetic anisotropy in the system. The observation of this phenomenon leads to open questions in the general understanding of magnetic heterostructures. Moreover, this shows that in general heterogeneous nanostructured materials may exhibit unexpected phenomena absent in the bulk. PMID- 16712026 TI - Interface free energies in p-spin glass models. AB - The replica method is used to calculate the interface free energy associated with the change from periodic to antiperiodic boundary conditions in finite dimensional p-spin glass models in the phase which at mean-field level has one step replica symmetry breaking (1RSB). In any finite dimension the interface free energy is exponentially small for a large system. This result implies that, in finite dimensions, the 1RSB state does not exist, as it is destroyed by thermal excitation of arbitrarily large droplets. The implications of this for the theory of structural glasses are discussed. PMID- 16712027 TI - Electron correlations and the minority-spin band gap in half-metallic Heusler alloys. AB - Electron-electron correlations affect the band gap of half-metallic ferromagnets by introducing nonquasiparticle states just above the Fermi level. In contrast with the spin-orbit coupling, a large asymmetric nonquasiparticle spectral weight is present in the minority-spin channel, leading to a peculiar finite-temperature spin depolarization effects. Using recently developed first-principle dynamical mean-field theory, we investigate these effects for the half-metallic ferrimagnetic Heusler compound FeMnSb. We discuss depolarization effects in terms of strength of local Coulomb interaction U and temperature in FeMnSb. We propose Ni(1-x)Fe(x)MnSb alloys as a perspective materials to be used in spin-valve structures and for experimental search of nonquasiparticle states in half metallic materials. PMID- 16712028 TI - Scenario for spin-glass phase with infinitely many states. AB - A possible phase in short-range spin glasses exhibiting infinitely many equilibrium states is proposed and characterized in real space. Experimental signatures in equilibrating systems measured with scanning probes are discussed. Some models with correlations in their exchange interactions are argued to exhibit this phase. Questions are raised about more realistic models and related issues. PMID- 16712030 TI - Electron spin precession upon reflecting from ferromagnetic surfaces. AB - Electrons with the polarization vector perpendicular to the magnetization of Fe, Co, and Ni films are spin analyzed after reflecting from the ferromagnet. At low primary electron energies a strong spin motion is found, namely, a precession of the polarization around the magnetization and a change of the angle between the polarization and the magnetization. This observation can be accounted for by the existence of spin-dependent gaps in the electronic band structure of the ferromagnets. PMID- 16712029 TI - Spiral spin order of self-assembled Co nanodisk arrays. AB - Spin order in hexagonal close packed cobalt nanodisk rows is quantitatively determined by off-axis electron holography. Periodic variation in the density of the local magnetic flux shows features of a spiral spin arrangement along the row axis, resulting from a tilted magnetic moment of nanoparticles with respect to the nanodisk axis. PMID- 16712031 TI - Quantum light emission of two lateral tunnel-coupled (In,Ga)As/GaAs quantum dots controlled by a tunable static electric field. AB - Lateral quantum coupling between two self-assembled (In,Ga)As quantum dots has been observed. Photon statistics measurements between the various excitonic and biexcitonic transitions of these lateral quantum dot molecules display strong antibunching confirming the presence of coupling. Furthermore, we observe an anomalous exciton Stark shift with respect to static electric field. A simple model indicates that the lateral coupling is due to electron tunneling between the dots when the ground states are in resonance. The electron probability can then be shifted to either dot and the system can be used to create a wavelength tunable single-photon emitter by simply applying a voltage. PMID- 16712032 TI - Valence band x-ray emission spectra of compressed germanium. AB - We report measurements of the valence band width in compressed Ge determined from x-ray emission spectra below the Ge K edge. The width of the valence band does not show any pressure dependence in the semiconducting diamond-type structure of Ge below 10 GPa. On the other hand, in the metallic beta-Sn phase above 10 GPa the valence band width increases under compression. Density-functional calculations show an increasing valence band width under compression both in the semiconducting phase (contrary to experiment) and in the metallic beta-Sn phase of Ge (in agreement with observed pressure-induced broadening). The pressure independent valence band width in the semiconducting phase of Ge appears to require theoretical advances beyond the density-functional theory or the GW approximation. PMID- 16712033 TI - Ultrafast Fiske effect in semiconductor superlattices. AB - The current flowing across a semiconductor superlattice in tilted electric and magnetic fields is known to exhibit resonant enhancement, when Landau states of neighboring wells align at certain ratios of the field strengths. We show that the ultrafast version of this effect, in which coherent electron wave packets are involved, has a profound analogy to the Fiske effect in superconductor Josephson junctions and superfluid weak links, in that the coupling of the tunneling induced charge oscillations (magneto-Bloch versus Josephson oscillations) to another oscillator (in-plane cyclotron oscillations versus external oscillator modes) opens an elastic rectifying transport channel. We explore the superlattice effect both theoretically and experimentally, and find that the transient self induced current can be adequately modeled if the damping of both types of coupled electron oscillations is properly taken into account. PMID- 16712034 TI - Polarization fluctuations in insulators and metals: new and old theories merge. AB - The ground-state fluctuation of polarization P is finite in insulators and divergent in metals, owing to the SWM sum rule [I. Souza, T. Wilkens, and R. M. Martin, Phys. Rev. B 62, 1666 (2000)]. This is a virtue of periodic (i.e., transverse) boundary conditions. I show that within any other boundary conditions the P fluctuation is finite even in metals, and a generalized sum rule applies. The boundary-condition dependence is a pure correlation effect, not present at the independent-particle level. In the longitudinal case inverted triangle x P = rho, and one equivalently addresses charge fluctuations: the generalized sum rule reduces then to a well-known result of the many-body theory. PMID- 16712035 TI - Electric-field-induced domain evolution in ferroelectric ultrathin films. AB - The electric-field-induced evolution of the recently discovered periodic 180 degree nanostripe domain structure is predicted in epitaxial Pb(Zr0.5Ti0.5)O3 ultrathin films from first principles. This evolution involves (1) the lateral growth of majority dipole domains at the expense of minority domains with the overall stripe periodicity remaining unchanged, (2) the creation of surface avoiding nanobubbles, and (3) the formation of a single monodomain state. Analogies and differences (i) with ferroelectric thin films made of BaTiO3 and (ii) with ferromagnetic thin films under magnetic field are discussed. PMID- 16712036 TI - Interface effects in ferroelectric PbTiO3 ultrathin films on a paraelectric substrate. AB - Interface effects on the ferroelectric behavior of PbTiO3 ultrathin films deposited on a SrTiO3 substrate are investigated using an interatomic potential approach with parameters fitted to first-principles calculations. We find that the correlation of atomic displacements across the film-substrate interface is crucial for the stabilization of the ferroelectric state in films a few unit cells thick. We show that the minimum film thickness for the appearance of a spontaneous polarized domain state is not an intrinsic property of the ferroelectric film but depends on the polarizability of the paraelectric substrate. We also observe that the substrate displays an induced polarization with an unusual oscillatory behavior. PMID- 16712037 TI - Magnetic double resonance in force microscopy. AB - Magnetic-resonance force microscopy is combined with cross-polarization and spin decoupling NMR techniques to obtain double-resonance NMR signals of micrometer scaled objects. The effective one-dimensional spatial resolution obtained in our experiments performed on a KPF6 single crystal sample is approximately 0.5 microm. The spectral linewidth of 900 Hz is sample limited. The described double resonance techniques can introduce new chemical specificity to the magnetic-force sensor. PMID- 16712038 TI - Elliptic phases: a study of the nonlinear elasticity of twist-grain boundaries. AB - We develop an explicit and tractable representation of a twist-grain-boundary phase of a smectic-A liquid crystal. This allows us to calculate the interaction energy between grain boundaries and the relative contributions from the bending and compression deformations. We discuss the special stability of the pi/2 grain boundaries and discuss the relation of this structure to the Schwarz D surface. PMID- 16712039 TI - Fluctuating velocity and momentum transfer in dense granular flows. AB - We show experimentally that in a free-surface granular flow the fluctuating velocity brings about momentum transfer at a considerable rate only very close to the free surface. Away from the free surface, where the flow is dense and stratified (or laminar), the fluctuating velocity plays no prominent dynamic role and stems passively from a kinematic constraint: The strata of particles must shake laterally as they slip past one another in the direction of the mean flow. Based on this insight, we formulate a simple model for the fluctuating velocity of dense granular flows. The predictions of the model agree well with our experimental measurements. PMID- 16712040 TI - Measuring inter-DNA potentials in solution. AB - Interactions between short strands of DNA can be tuned from repulsive to attractive by varying solution conditions and have been quantified using small angle x-ray scattering techniques. The effective DNA interaction charge was extracted by fitting the scattering profiles with the generalized one-component method and inter-DNA Yukawa pair potentials. A significant charge is measured at low to moderate monovalent counterion concentrations, resulting in strong inter DNA repulsion. The charge and repulsion diminish rapidly upon the addition of divalent counterions. An intriguing short range attraction is observed at surprisingly low divalent cation concentrations, approximately 16 mM Mg2+. Quantitative measurements of inter-DNA potentials are essential for improving models of fundamental interactions in biological systems. PMID- 16712041 TI - Stability and growth of single myelin figures. AB - Myelin figures are long thin cylindrical structures that typically grow as a dense tangle when water is added to the concentrated lamellar phase of certain surfactants. We show that, starting from a well-ordered initial state, single myelin figures can be produced in isolation thus allowing a detailed study of their growth and stability. These structures grow with their base at the exposed edges of bilayer stacks from which material is transported into the myelin. Myelins only form and grow in the presence of a driving stress; when the stress is removed, the myelins retract. PMID- 16712042 TI - Yielding and flow in adhesive and nonadhesive concentrated emulsions. AB - The nonlinear rheological response of soft glassy materials is addressed experimentally by focusing on concentrated emulsions where interdroplet attraction is tuned through varying the surfactant content. Velocity profiles are recorded using ultrasonic velocimetry simultaneously to global rheological data in the Couette geometry. Our data show that nonadhesive and adhesive emulsions have radically different flow behaviors in the vicinity of yielding: while the flow remains homogeneous in the nonadhesive emulsion and the Herschel-Bulkley model for a yield stress fluid describes the data very accurately, the adhesive system displays shear localization and does not follow a simple constitutive equation, suggesting that the mechanisms involved in yielding transitions are not universal. PMID- 16712043 TI - Many-body hydrodynamic interactions in charge-stabilized suspensions. AB - In this joint experimental-theoretical work we study hydrodynamic interaction effects in dense suspensions of charged colloidal spheres. Using x-ray photon correlation spectroscopy we have determined the hydrodynamic function H(q), for a varying range of electrosteric repulsion. We show that H(q) can be quantitatively described by means of a novel Stokesian dynamics simulation method for charged Brownian spheres, and by a modification of a many-body theory developed originally by Beenakker and Mazur. Very importantly, we can explain the behavior of H(q) for strongly correlated particles without resorting to the controversial concept of hydrodynamic screening, as was attempted in earlier work by Riese [Phys. Rev. Lett. 85, 5460 (2000)]. PMID- 16712044 TI - Disordered network state in hydrated block-copolymer surfactants. AB - Hydration of poly(butadiene-b-ethylene oxide) diblock copolymers leads to various ordered and disordered phases, analogous to the aqueous phase behavior of surfactants and lipids. Small-angle x-ray scattering measurements corroborated by cryogenic scanning electron microscopy imaging reveal a random network (N) morphology at polymer compositions and water content intermediate to those associated with ordered cylinders (H1) and lamellae (L). This sequence of self assembled structures is strikingly similar to the phase behavior of certain water oil-surfactant microemulsions. PMID- 16712045 TI - Ultrafast vibrational and structural dynamics of the proton in liquid water. AB - The dynamical behavior of excess protons in liquid water is investigated using femtosecond vibrational pump-probe spectroscopy. By resonantly exciting the O-H+ stretching mode of the H9O4(+) (Eigen) hydration structure of the proton and probing the subsequent absorption change over a broad frequency range, the dynamics of the proton is observed in real time. The lifetime of the protonic stretching mode is found to be approximately 120 fs, shorter than for any other vibration in liquid water. We also observe the interconversion between the H9O4(+) (Eigen) and H5O2(+) (Zundel) hydration structures of the proton. This interconversion, which constitutes an essential step of proton transport in water, is found to occur on an extremely fast (< 100 fs) time scale. PMID- 16712046 TI - Confinement-induced novel morphologies of block copolymers. AB - Self-assembly of block copolymers confined in cylindrical nanopores is studied systematically using a simulated annealing technique. For diblock copolymers which form two-dimensional hexagonally packed cylinders with period L0 in the bulk, novel structures such as helices and stacked toroids spontaneously form inside the cylindrical pores. These confinement-induced morphologies have no counterpart in the bulk system and they depend on the pore diameter (D) and the surface-polymer interactions, reflecting the importance of structural frustration and interfacial interactions. On tightening the degree of confinement, transitions from helices to toroids to spheres are observed. Mechanisms of the morphological transitions can be understood based on the degree of structural frustration parametrized by the ratio D/L0. PMID- 16712047 TI - High-frequency stress relaxation in semiflexible polymer solutions and networks. AB - We measure the linear viscoelasticity of sterically entangled and chemically cross-linked networks of actin filaments over more than five decades of frequency. The high-frequency response reveals rich dynamics unique to semiflexible polymers, including a previously unobserved relaxation due to rapid axial tension propagation. For high molecular weight, and for cross-linked gels, we obtain quantitative agreement with predicted shear moduli in both amplitude and frequency dependence. PMID- 16712048 TI - Prediction and observation of crystal structures of oppositely charged colloids. AB - We studied crystal structures in mixtures of large and small oppositely charged spherical colloids with size ratio 0.31 using Monte Carlo simulations and confocal microscopy. We developed an interactive method based on simulated annealing to predict new binary crystal structures with stoichiometries from 1 to 8. Employing these structures in Madelung energy calculations using a screened Coulomb potential, we constructed a ground-state phase diagram, which shows a remarkably rich variety of crystals. Our phase diagram displays colloidal analogs of simple-salt structures and of the doped fullerene C60 structures, but also novel structures that do not have an atomic or molecular analog. We found three of the predicted structures experimentally, which provides confidence that our method yields reliable results. PMID- 16712049 TI - Layered complex networks. AB - Many complex networks are only a part of larger systems, where a number of coexisting topologies interact and depend on each other. We introduce a layered model to facilitate the description and analysis of such systems. As an example of its application, we study the load distribution in three transportation systems, where the lower layer is the physical infrastructure and the upper layer represents the traffic flows. This layered view allows us to capture the fundamental differences between the real load and commonly used load estimators, which explains why these estimators fail to approximate the real load. PMID- 16712050 TI - Comment on "Projective quantum Monte Carlo method for the Anderson impurity model and its application to dynamical mean field theory". PMID- 16712052 TI - Bose-Einstein condensation with an entangled order parameter. AB - We propose a practically accessible non-mean-field ground state of Bose-Einstein condensation, which occurs in an interspecies two-particle entangled state, and is thus described by an entangled order parameter. A suitably defined entanglement entropy is used as the characterization of the non-mean-field nature, and is found to persist in a wide parameter regime. The interspecies entanglement leads to novel interference terms in the dynamical equations governing the single-particle orbital wave function. Experimental feasibility and several methods of probe are discussed. We urge the study of multichannel scattering between different species of atoms. PMID- 16712053 TI - Pair production and optical lasers. AB - Electron-positron pair creation in a standing wave is explored using a parameter free quantum kinetic equation. Field strengths and frequencies corresponding to modern optical lasers induce a material polarization of the QED vacuum, which may be characterized as a plasma of e+e- quasiparticle pairs with a density of approximately 10(20) cm-3. The plasma vanishes almost completely when the laser field is zero, leaving a very small residual pair density n(r) which is the true manifestation of vacuum decay. The average pair density per period is proportional to the laser intensity but independent of the frequency nu. The density of residual pairs also grows with laser intensity but n(r) proportional to nu(2). With optical lasers at the forefront of the current generation, these dynamical QED vacuum effects can plausibly generate 5-10 observable two-photon annihilation events per laser pulse. PMID- 16712054 TI - Achieving peak brightness in an atom laser. AB - In this Letter we present experimental results and a simple analytic theory on the first continuous (long pulse) Raman atom laser. We analyze the flux and brightness of a generic two state atom laser with an analytic model that shows excellent agreement with our experiments. We show that, for the same source size, the brightness achievable with a Raman atom laser is at least 3 orders of magnitude greater than achievable in any other demonstrated continuously outcoupled atom laser. PMID- 16712055 TI - Strong-coupling theory for the superfluidity of Bose-Fermi mixtures. AB - We develop a strong-coupling theory for the superfluidity of fermion pairing phase in a Bose-Fermi mixture. Dynamical screening, self-energy renormalization, and a pairing gap function are included self-consistently within the adiabatic limit (i.e., the phonon velocity is much smaller than the Fermi velocity). An analytical solution for the transition temperature (T(c)) is derived within reasonable approximations. Using typical parameters of a 40K-87Rb mixture, we find that the calculated T(c) is several times larger than that obtained in the weak coupling theory, and can be up to several percent of the Fermi temperature. PMID- 16712056 TI - Change of decoherence scenario and appearance of localization due to reservoir anharmonicity. AB - Although coupling to a super-Ohmic bosonic reservoir leads only to partial dephasing on short time scales, exponential decay of coherence appears in the Markovian limit (for long times) if anharmonicity of the reservoir is taken into account. This effect not only qualitatively changes the decoherence scenario but also leads to localization processes in which superpositions of spatially separated states dephase with a rate that depends on the distance between the localized states. As an example of the latter process, we study the decay of coherence of an electron state delocalized over two semiconductor quantum dots due to anharmonicity of phonon modes. PMID- 16712057 TI - Phase winding in chiral liquid crystalline monolayers due to Lehmann effects. AB - We model recent experimental results by Tabe and Yokoyama on chiral liquid crystalline monolayers driven by the transmembrane transfer of water molecules. We point out that the target patterns found for the orientation of the in-plane preferred direction can be described as phase winding due to Lehmann effects, which are specific to macroscopically chiral systems. The model presented has only one variable and is driven and dissipative. Including the effects of noise leads to an accurate representation of the patterns observed experimentally. Future experimental tests of the model are suggested. PMID- 16712058 TI - Identifying diffusion processes in one-dimensional lattices in thermal equilibrium. AB - In this Letter, I propose that a properly rescaled spatiotemporal correlation function of the energy density fluctuations may be applied to characterize the equilibrium diffusion processes in lattice systems with finite temperature. Applying this function, the diffusion processes in three one-dimensional nonlinear lattices are studied. The diffusion exponent is shown to be related to the diverging exponent of the thermal conductivity of a lattice through the relation , as has been proved based on the Levy walk assumption. The diffusion behavior is explained in terms of solitons and phonons. PMID- 16712059 TI - Universality away from critical points in two-dimensional phase transitions. AB - The p-state clock model in two dimensions is a system of discrete rotors with a quasiliquid phase in a region T14. We show that, for p>4 and above a temperature T(eu), all macroscopic thermal averages become identical to those of the continuous rotor (p=infinity). This collapse of thermodynamic observables creates a regime of extended universality in the phase diagram and an emergent symmetry, not present in the Hamiltonian. For p> or =8, the collapse starts in the quasiliquid phase and makes the transition at T2 identical to the Berezinskii Kosterlitz-Thouless (BKT) transition of the continuous rotor. For p< or =6, the transition at T2 is below T(eu) and no longer a BKT transition. The results generate a range of experimental predictions, such as the motion of magnetic domain walls, and limits on macroscopic distinguishability of different microscopic interactions. PMID- 16712060 TI - Decay of Loschmidt echo enhanced by quantum criticality. AB - We study the transition of a quantum system from a pure state to a mixed one, which is induced by the quantum criticality of the surrounding system E coupled to it. To characterize this transition quantitatively, we carefully examine the behavior of the Loschmidt echo (LE) of E modeled as an Ising model in a transverse field, which behaves as a measuring apparatus in quantum measurement. It is found that the quantum critical behavior of E strongly affects its capability of enhancing the decay of LE: near the critical value of the transverse field entailing the happening of quantum phase transition, the off diagonal elements of the reduced density matrix describing S vanish sharply. PMID- 16712061 TI - Quantum nature of the big bang. AB - Some long-standing issues concerning the quantum nature of the big bang are resolved in the context of homogeneous isotropic models with a scalar field. Specifically, the known results on the resolution of the big-bang singularity in loop quantum cosmology are significantly extended as follows: (i) the scalar field is shown to serve as an internal clock, thereby providing a detailed realization of the "emergent time" idea; (ii) the physical Hilbert space, Dirac observables, and semiclassical states are constructed rigorously; (iii) the Hamiltonian constraint is solved numerically to show that the big bang is replaced by a big bounce. Thanks to the nonperturbative, background independent methods, unlike in other approaches the quantum evolution is deterministic across the deep Planck regime. PMID- 16712062 TI - String splitting and strong coupling meson decay. AB - We study the decay of high spin mesons using the gauge-string theory correspondence. The rate of the process is calculated by studying the splitting of a macroscopic string intersecting a D-brane. The result is applied to the decay of mesons in N=4 super Yang-Mills theory with a small number of flavors and in a gravity dual of large N QCD. In QCD the decay of high spin mesons is found to be heavily suppressed in the regime of validity of the supergravity description. PMID- 16712063 TI - Enhanced electroweak penguin amplitude in B-->VV decays. AB - We discuss a novel electromagnetic penguin contribution to the transverse helicity amplitudes in B decays to two vector mesons, which is enhanced by two powers of mB/Lambda relative to the standard penguin amplitudes. This leads to unique polarization signatures in penguin-dominated decay modes such as B-->rhoK* similar to polarization effects in the radiative decay B-->K*gamma and offers new opportunities to probe the magnitude and chirality of flavor-changing neutral current couplings to photons. PMID- 16712064 TI - Probing MeV dark matter at low-energy e+e- colliders. AB - It has been suggested that the pair annihilation of dark matter particles chi with mass between 0.5 and 20 MeV into e+e- pairs could be responsible for the excess flux (detected by the INTEGRAL satellite) of 511 keV photons coming from the central region of our Galaxy. The simplest way to achieve the required cross section while respecting existing constraints is to introduce a new vector boson U with mass M(U) below a few hundred MeV. We point out that over most of the allowed parameter space, the process e+e--->U(gamma), followed by the decay of U into either an e+e- pair or an invisible (nu(-)nu or chi(-)chi) channel, should lead to signals detectable by current B-factory experiments. A smaller, but still substantial, region of parameter space can also be probed at the Phi factory DAPhiNE. PMID- 16712065 TI - Nuclear quantum optics with x-ray laser pulses. AB - The direct interaction of nuclei with superintense laser fields is studied. We show that present and upcoming high-frequency laser facilities, especially together with a moderate acceleration of the target nuclei to match photon and transition frequency, do allow for resonant laser-nucleus interaction. These direct interactions may be utilized for the model-independent optical measurement of nuclear properties such as the transition frequency and the dipole moment, thus opening the field of nuclear quantum optics. As an ultimate goal, one may hope that direct laser-nucleus interactions could become a versatile tool to enhance preparation, control, and detection in nuclear physics. PMID- 16712066 TI - Neutrino-induced nucleosynthesis of A>64 nuclei: the nu p process. AB - We present a new nucleosynthesis process that we denote as the nu p process, which occurs in supernovae (and possibly gamma-ray bursts) when strong neutrino fluxes create proton-rich ejecta. In this process, antineutrino absorptions in the proton-rich environment produce neutrons that are immediately captured by neutron-deficient nuclei. This allows for the nucleosynthesis of nuclei with mass numbers A>64, , making this process a possible candidate to explain the origin of the solar abundances of (92,94)Mo and (96,98)Ru. This process also offers a natural explanation for the large abundance of Sr seen in a hyper-metal-poor star. PMID- 16712067 TI - Enhancement and control of H2 dissociative ionization by femtosecond VUV laser pulses. AB - We report ab initio calculations of H2 ionization by VUV/fs 10(12) W/cm2 laser pulses including correlation and all electronic and vibrational degrees of freedom (DOF). Inclusion of the nuclear DOF leads to a substantial increase of resonance enhanced multiphoton ionization. By varying pulse duration, it is possible to control the ratio of dissociative to nondissociative ionization as well as the final H+(2) vibrational distribution. For pulses longer than 10 fs and proportional to omega>0.46 a.u., dissociative ionization entirely dominates, which is a very unusual situation in photoionization studies. PMID- 16712068 TI - Direct experimental probe of the on-site Coulomb repulsion in the doubly charged fullerene anion C70 2-. AB - Vibrationally resolved photoelectron spectra were obtained for cold C(70)- and C(70)2-. Accurate values for the first and second electron affinities (EA's) of C(70) were measured as 2.765 +/- 0.010 and 0.002 (+0.01/-0.03)eV, respectively, establishing that C(70)-2 is an electronically stable dianion in the gas phase. The difference between the first and second EA (2.75 eV) provides a direct experimental measure for the on-site Coulomb and exchange interactions between the two excess electrons in C(70)-2. Strong electron correlation effects were also observed between the two excess electrons in C(70)-2. PMID- 16712069 TI - Interference oscillations in the angular distribution of laser-ionized electrons near ionization threshold. AB - We analyze the two-dimensional momentum distribution of electrons ionized by few cycle laser pulses in the transition regime from multiphoton absorption to tunneling by solving the time-dependent Schrodinger equation and by a classical trajectory Monte-Carlo simulation with tunneling (CTMC-T). We find a complex two dimensional interference pattern that resembles above threshold ionization (ATI) rings at higher energies and displays Ramsauer-Townsend-type diffraction oscillations in the angular distribution near threshold. CTMC-T calculations provide a semiclassical explanation for the dominance of selected partial waves. While the present calculation pertains to hydrogen, we find surprising qualitative agreement with recent experimental data for rare gases [A. Rudenko, J. Phys. B 37, L407 (2004)]. PMID- 16712070 TI - Cold molecule spectroscopy for constraining the evolution of the fine structure constant. AB - We report precise measurements of ground-state, Lambda-doublet microwave transitions in the hydroxyl radical molecule (OH). Utilizing slow, cold molecules produced by a Stark decelerator we have improved over the precision of the previous best measurement 25-fold for the F'=2-->F=2 transition, yielding (1 667 358 996 +/- 4)Hz, and by tenfold for the F'=1-->F=1 transition, yielding (1 665 401 803 +/-12)Hz. Comparing these laboratory frequencies to those from OH megamasers in interstellar space will allow a sensitivity of 1 ppm for Delta(alpha/alpha) over approximately 10(10) yr. PMID- 16712071 TI - Laser cooling without repumping: a magneto-optical trap for erbium atoms. AB - We report on a novel mechanism that allows for strong laser cooling of atoms that do not have a closed cycling transition. This mechanism is observed in a magneto optical trap (MOT) for erbium, an atom with a very complex energy level structure with multiple pathways for optical-pumping losses. We observe surprisingly high trap populations of over 10(6) atoms and densities of over 10(11) atoms cm(-3), despite the many potential loss channels. A model based on recycling of metastable and ground state atoms held in the quadrupole magnetic field of the trap explains the high trap population, and agrees well with time-dependent measurements of MOT fluorescence. The demonstration of trapping of a rare-earth atom such as erbium opens a wide range of new possibilities for practical applications and fundamental studies with cold atoms. PMID- 16712073 TI - Dispersion spreading of biphotons in optical fibers and two-photon interference. AB - We present the first observation of two-photon polarization interference structure in the second-order Glauber correlation function of two-photon light generated via type-II spontaneous parametric down-conversion. In order to obtain this result, two-photon light is transmitted through an optical fiber and the coincidence distribution is analyzed by means of the start-stop method. Beyond the experimental demonstration of an interesting effect in quantum optics, these results also have considerable relevance for quantum communications. PMID- 16712072 TI - Saturated ionization of fullerenes in intense laser fields. AB - We investigate the ionization of icosahedral fullerenes (C20, C60, C80, and C180) in an intense laser pulse using the S-matrix theory. The results obtained are in excellent agreement with the recent observations of unexpectedly high saturation intensities of the Buckminster fullerene and its multiply charged ions. Our analysis strongly suggests that the related phenomenon of suppressed ionization of these complex fullerenes is due to the finite cage size and the "multislit" interference effect between partial waves emitted from the different nuclei rather than to a dynamical multielectron polarization effect. PMID- 16712074 TI - Direct observation of optical precursors in a region of anomalous dispersion. AB - We create optical precursors by propagating a step-modulated optical pulse through a linear resonant dielectric absorber. The field emerging from the dielectric consists of a several-nanosecond-long spike with near 100% transmission, which decays to a constant value expected from Beer's law. This high-transmission spike might be useful for imaging applications requiring penetrating optical radiation. We compare our observations to two different theories, revealing that the spike consists of both the Sommerfeld and Brillouin precursors. PMID- 16712075 TI - Longitudinal optical binding of high optical contrast microdroplets in air. AB - Binding along the beam axis (which we shall call "longitudinal optical binding") has been observed between micron-sized oil droplets in a three dimensional optical trap in air. We argue that it is the high optical contrast which is responsible for the exceptionally stable doublet structures observed experimentally. It was also observed that optically bound doublets tend to cling to interference fringes created by the two counterpropagating beams. Our observations are qualitatively discussed in the context of both the ray model (optics) approximation, and in the Rayleigh (dipolar) range. Our observations were consistent with calculations of binding and trapping forces which we carried out by employing an exact multiple-scattering theory. PMID- 16712076 TI - Semiconductor nanowire ring resonator laser. AB - Nanowires of the wide band-gap semiconductor gallium nitride (GaN) have been shown to act as room-temperature uv lasers. Recent advances in nanomanipulation have made it possible to modify the shape of these structures from a linear to a pseudoring conformation. Changes to the optical boundary conditions of the lasing cavity affect the structure's photoluminescence, photon confinement, and lasing as a function of ring diameter. For a given cavity, ring-mode redshifting is observed to increase with decreasing ring diameter. Significant shifts, up to 10 nm for peak emission values, are observed during optical pumping of a ring resonator nanolaser compared to its linear counterpart. The shifting appears to result from conformational changes of the cavity rather than effects such as band gap renormalization, allowing the mode spacing and position to be tuned with the same nanowire gain medium. PMID- 16712077 TI - Spontaneous emergence of periodic patterns in a biologically inspired simulation of photonic structures. AB - We simulate an evolutionary process in the lab for designing a novel high confinement photonic structure, starting with a set of completely random patterns, with no insight on the initial geometrical pattern. We show a spontaneous emergence of periodical patterns as well as previously unseen high confinement subwavelength bowtie regions. The evolved structure has a Q of 300 and an ultrasmall modal volume of 0.112 (lambda/2n)3. The emergence of the periodic patterns in the structure indicates that periodicity is a principal condition for effective control of the distribution of light. PMID- 16712078 TI - Coarse graining the dynamics of coupled oscillators. AB - We present an equation-free computational approach to the study of the coarse grained dynamics of finite assemblies of nonidentical coupled oscillators at and near full synchronization. We use coarse-grained observables which account for the (rapidly developing) correlations between phase angles and natural frequencies. Exploiting short bursts of appropriately initialized detailed simulations, we circumvent the derivation of closures for the long-term dynamics of the assembly statistics. PMID- 16712079 TI - Making forecasts for chaotic physical processes. AB - Making a prediction for a chaotic physical process involves specifying the probability associated with each possible outcome. Ensembles of solutions are frequently used to estimate this probability distribution. However, for a typical chaotic physical system and model of that system, no solution of remains close to for all time. We propose an alternative. This Letter shows how to inflate or systematically perturb the ensemble of solutions of so that some ensemble member remains close to for orders of magnitude longer than unperturbed solutions of . This is true even when the perturbations are significantly smaller than the model error. PMID- 16712080 TI - Monodisperse drop formation in square microchannels. AB - The electrohydrodynamic instability of the interface between two liquids with different physical and electrical properties in plane Poiseuille flow is used to form monodisperse droplets in a square channel. The drop size and formation rate are controlled by simply controlling the flow rates and the amplitude of the electric field applied across the channel. PMID- 16712081 TI - Elastic instabilities of polymer solutions in cross-channel flow. AB - When polymer molecules pass near the hyperbolic point of a microchannel cross flow, they are strongly stretched. As the strain rate is varied at low Reynolds number (< 10(-2)), tracer and particle-tracking experiments show that molecular stretching produces two flow instabilities: one in which the velocity field becomes strongly asymmetric, and a second in which it fluctuates nonperiodically in time. The flow is strongly perturbed even far from the region of instability, and this phenomenon can be used to produce mixing. PMID- 16712082 TI - Fully coherent x-ray pulses from a regenerative-amplifier free-electron laser. AB - We propose and analyze a regenerative-amplifier free-electron laser (FEL) to produce fully coherent, hard x-ray pulses. The method makes use of narrow bandwidth Bragg crystals to form an x-ray feedback loop around a relatively short undulator. Self-amplified spontaneous emission (SASE) from the leading electron bunch in a bunch train is spectrally filtered by the Bragg reflectors and is brought back to the beginning of the undulator to interact repeatedly with subsequent bunches in the bunch train. The FEL interaction with these short bunches regeneratively amplifies the radiation intensity and broadens its spectrum, allowing for effective transmission of the x rays outside the crystal bandwidth. The spectral brightness of these x-ray pulses is about 2 to 3 orders of magnitude higher than that from a single-pass SASE FEL. PMID- 16712083 TI - Role of turbulence on edge momentum redistribution in the TJ-II stellarator. AB - Radial profiles of the parallel-radial Reynolds stress component, proportional to the cross correlation between parallel and radial fluctuating velocities, have been measured in the plasma boundary region of the TJ-II stellarator. Experimental results show the existence of significant parallel turbulent forces at plasma densities above the threshold value to trigger perpendicular sheared flows. This finding provides the first experimental evidence of the role of parallel turbulence forces on edge momentum redistribution in fusion devices. PMID- 16712084 TI - Displacement current and the generation of parallel electric fields. AB - We show for the first time the dynamical relationship between the generation of magnetic field-aligned electric field (E||) and the temporal changes and spatial gradients of magnetic and velocity shears, and the plasma density in Earth's magnetosphere. We predict that the signatures of reconnection and auroral particle acceleration should have a correlation with low plasma density, and a localized voltage drop (V||) should often be associated with a localized magnetic stress concentration. Previous interpretations of the E|| generation are mostly based on the generalized Ohm's law, causing serious confusion in understanding the nature of reconnection and auroral acceleration. PMID- 16712085 TI - Shear viscosity and shear thinning in two-dimensional Yukawa liquids. AB - A two-dimensional Yukawa liquid is studied using two different nonequilibrium molecular dynamics simulation methods. Shear viscosity values in the limit of small shear rates are reported for a wide range of Coulomb coupling parameter and screening lengths. At high shear rates it is demonstrated that this liquid exhibits shear thinning; i.e., the viscosity eta diminishes with increasing shear rate. It is expected that two-dimensional dusty plasmas will exhibit this effect. PMID- 16712086 TI - Cause of sudden magnetic reconnection in a laboratory plasma. AB - The cause for sudden reconnection in reversed field pinch plasmas is determined experimentally for two cases: large reconnection events (the sawtooth crash) and small reconnection events during improved confinement. We measure the term in the MHD equations which represents the driving (or damping) of edge tearing modes due to the axisymmetric magnetic field. The term is negative for large reconnection events (the modes are stable, implying that reconnection may be driven by nonlinear coupling to other modes) and positive for small reconnection events (modes are unstable, reconnection is spontaneous). PMID- 16712087 TI - Numerical simulations of type-III solar radio bursts. AB - The first numerical simulations are presented for type-III solar radio bursts in the inhomogeneous solar corona and interplanetary space, that include microscale quasilinear and nonlinear processes, intermediate-scale driven ambient density fluctuations, and large scale evolution of electron beams, Langmuir and ion sound waves, and fundamental and harmonic electromagnetic emission. Bidirectional coronal emission is asymmetric between the upward and downward directions, and harmonic emission dominates fundamental emission. In interplanetary space, fundamental and/or harmonic emission can be important. Langmuir and ion sound waves are bursty and the statistics of Langmuir wave energy agree well with the predictions of stochastic growth theory. PMID- 16712088 TI - Quasimonoenergetic deuteron bursts produced by ultraintense laser pulses. AB - We report on the generation and laser acceleration of bunches of energetic deuterons with a small energy spread at about 2 MeV. This quasimonoenergetic peak within the ion energy spectrum was observed when heavy-water microdroplets were irradiated with ultrashort laser pulses of about 40 fs duration and high (10(-8)) temporal contrast, at an intensity of 10(19) W/cm(2). The results can be explained by a simple physical model related to spatial separation of two ion species within a finite-volume target. The production of quasimonoenergetic ions is a long-standing goal in laser-particle acceleration; it could have diverse applications such as in medicine or in the development of future compact ion accelerators. PMID- 16712089 TI - Identification of the carbon antisite-vacancy pair in 4H-SiC. AB - The metastability of vacancies was theoretically predicted for several compound semiconductors alongside their transformation into the antisite-vacancy pair counterpart; however, no experiment to date has unambiguously confirmed the existence of antisite-vacancy pairs. Using electron paramagnetic resonance and first principles calculations we identify the S15 center as the carbon antisite vacancy pair in the negative charge state (C(Si)V-(C)) in 4H-SiC. We suggest that this defect is a strong carrier-compensating center in n-type or high-purity semi insulating SiC. PMID- 16712090 TI - Stokes-Einstein relation in supercooled aqueous solutions of glycerol. AB - The diffusion of glycerol molecules decreases with decreasing temperature as its viscosity increases in a manner simply described by the Stokes-Einstein relation. Approaching the glass transition, this relation breaks down as it does with a number of other pure liquid glass formers. We have measured the diffusion coefficient for binary mixtures of glycerol and water and find that the Stokes Einstein relation is restored with increasing water concentration. Our comparison with theory suggests that adding water postpones the formation of frustration domains. PMID- 16712091 TI - Pressure-induced lock-in in an aperiodic nanoporous crystal. AB - This Letter reports on the first observation of a commensurate lock-in inside an aperiodic composite. This result is obtained by neutron diffraction, under hydrostatic pressure, in the prototype self-assembled crystal of hexadecane urea. A selective compressibility of the sublattices is a required condition together with the existence of a lock-in energy term in these supramolecular materials. This measurement, under one-dimensional confinement, opens the way for the use of continuously controlled forces to manipulate molecular properties or functions. PMID- 16712092 TI - Correlation between floppy to rigid transitions and non-Arrhenius conductivity in glasses. AB - Non-Arrhenius behavior is observed for a number of virgin potassium silicate glasses (1-x)SiO2-xK2O with a potassium oxide concentration larger than a certain value x = x(c). Recovering of Arrhenius behavior is provided by the annealing that enhances densification. These various results are the manifestation of the floppy or rigid nature of the network. Compositional effects and saturation with temperature can be analyzed with a combination of constraint theory and a point defect model. They underscore the key role played by network rigidity for the understanding of conduction and saturation effects in glassy electrolytes. PMID- 16712093 TI - Role of interparticle forces in the formation of random loose packing. AB - We present a physical and numerical study of the settling of uniform spheres in liquids and show that interparticle forces play a critical role in forming the so called random loose packing (RLP). Different packing conditions give different interparticle forces and, hence, different RLP. Two types of interparticle forces are identified: process dependent and process independent. The van der Waals force, as the major cohesive force in the present study, plays a critical role in effecting the process-dependent forces such as drag and lift forces. An equation is formulated to describe the relationship between the macroscopic packing fraction and microscopic interparticle forces in a packing. We argue there is no lowest packing fraction for a mechanically stable RLP; hence, the packing fractions of RLP can range from 0 to 0.64 depending on the cohesive and frictional conditions between particles. PMID- 16712094 TI - Mechanical behavior of bulk amorphous alloys reinforced by ductile particles at cryogenic temperatures. AB - The mechanical behavior of Zr-based bulk amorphous alloy composites (BAACs) was investigated at 77 K. The 5 vol. % Ta-BAAC maintained large plastic strains of approximately 13% with a 16% strength increase, when compared with that at 298 K. The interaction between shear bands and particles shows that shear extension in particles has limited penetration, and shear bands build up around particles. In addition to on the failure surface of the amorphous matrix, molten characteristics were also found on the surface of sheared particles. Pair distribution function studies were performed to understand the mechanical behavior. PMID- 16712095 TI - Universality of synchrony: critical behavior in a discrete model of stochastic phase-coupled oscillators. AB - We present the simplest discrete model to date that leads to synchronization of stochastic phase-coupled oscillators. In the mean field limit, the model exhibits a Hopf bifurcation and global oscillatory behavior as coupling crosses a critical value. When coupling between units is strictly local, the model undergoes a continuous phase transition which we characterize numerically using finite-size scaling analysis. In particular, the onset of global synchrony is marked by signatures of the XY universality class, including the appropriate classical exponents beta and nu, a lower critical dimension d(lc) = 2, and an upper critical dimension d(uc) = 4. PMID- 16712096 TI - Signatures of glass formation in a fluidized bed of hard spheres. AB - We demonstrate that a fluidized bed of hard spheres during defluidization displays properties associated with formation of a glass. The final state is rate dependent, and as this state is approached, the bed exhibits heterogeneity with increasing time and length scales. The formation of a glass results in the arrest of macroscopic particle motion and thus the loss of fluidization. Microscopic motion persists in this state, but the bed can be jammed by application of a small increase in flow rate. Thus a fluidized bed can serve as a test system for studies of glass formation and jamming. PMID- 16712097 TI - Revisiting the structure of the p(4 x 4) surface oxide on Ag(111). AB - Scanning tunneling microscopy (STM) and density-functional theory are used to reexamine the structure of the renowned p(4 x 4)-O/Ag(111) surface oxide. The accepted structural model [C. I. Carlisle, Phys. Rev. Lett. 84, 3899 (2000)10.1103/PhysRevLett.84.3899] is incompatible with the enhanced resolution of the current STM measurements. An "Ag6 model" is proposed that is more stable than its predecessor and accounts for the coexistence of the p(4 x 4) and a novel c(3 x 5log3)rect phase. This coexistence is an indication of the dynamic complexity of the system that until now has not been appreciated. PMID- 16712098 TI - Structure of Ag(111)-p(4 x 4)-O: no silver oxide. AB - The structure of the oxygen-induced p(4 x 4) reconstruction of Ag(111) is determined by a combination of scanning tunneling microscopy, surface x-ray diffraction, core level spectroscopy, and density functional theory. We demonstrate that all previous models of this surface structure are incorrect and propose a new model which is able to explain all our experimental findings but has no resemblance to bulk silver oxide. We also shed some light on the limitations of current density functional theories and the potential role of van der Waals interactions in the stabilization of oxygen-induced surface reconstructions of noble metals. PMID- 16712099 TI - Instabilities and coarsening of stressed crystal surfaces in aqueous solution. AB - Strong pattern formation occurs on polished miscut surfaces of sodium chlorate (NaClO3) single crystals that are uniaxially stressed perpendicular to the step edge direction and placed in a saturated aqueous solution. The wavelength lambda of the stress-induced surface instability increased continuously in experiments up to 9 days after placed in the solution. There were three successive regimes of coarsening: (i) one-dimensional step bunching with lambda approximately t(1/4) until an undulation transition was reached, (ii) a two-dimensional coarsening mechanism with lambda approximately t(1/2), and a gradual transition to (iii) Ostwald ripening-like coarsening with lambda approximately t(1/3). The coarsening of the surface patterns towards a stable, flat surface implies the spontaneous formation of a stress-free skin on the surface of the stressed solid. PMID- 16712100 TI - Physisorption-induced C-H bond elongation in methane. AB - Physisorption of methane to a Pt surface was studied by x-ray absorption spectroscopy in combination with density functional theory spectrum calculations. The experiment shows new electronic states appearing upon physisorption. We find that these states are due to orbital mixing causing charge polarization as a means to minimize Pauli repulsion. The results can be explained by elongation of 1 C-H bond by 0.09 A in the physisorbed state even though no covalent chemical bond is formed. PMID- 16712101 TI - Breakdown of dynamic scaling in surface growth under shadowing. AB - Using Monte Carlo simulations and experimental results, we show that for common thin film deposition techniques, such as sputter deposition and chemical vapor deposition, a mound structure can be formed with a characteristic length scale, or "wavelength" lambda, that describes the separation of the mounds. We show that the temporal evolution of lambda is distinctly different from that of the mound size, or lateral correlation length xi. The formation of a mound structure is due to nonlocal growth effects, such as shadowing, that lead to the breakdown of the self-affinity of the morphology described by the well-established dynamic scaling theory. We show that the wavelength grows as a function of time in a power law form, lambda approximately t(p), where p approximately equals 0.5 for a wide range of growth conditions, while the mound size grows as xi approximately t(1 z), where 1/z varies depending on growth conditions. PMID- 16712102 TI - Wettability control of droplet deposition and detachment. AB - The conditions for droplet deposition on plane substrates are studied using electrowetting to continuously modulate the surface wettability. Droplets of controlled volume attached to the tip of a pipette are brought into contact with the surface. During retraction of the pipette the droplets are deposited or detach completely depending on volume and contact angle. The experimental limit of deposition in the contact angle or volume plane is in good agreement with analytical and numerical predictions obtained within the capillary model. PMID- 16712103 TI - Application of van der Waals density functional to an extended system: adsorption of benzene and naphthalene on graphite. AB - It is shown that it is now possible to include van der Waals (vdW) interactions via a nonempirical implementation of density functional (DF) theory to describe the correlation energy in electronic structure calculations on infinite systems of no particular symmetry. The vdW-DF theory [Phys. Rev. Lett. 92, 246401 (2004)] is applied to the adsorption of benzene and naphthalene on an infinite sheet of graphite, as well as the binding between two graphite sheets. A comparison with recent thermal-desorption data [Phys. Rev. B 69, 155406 (2004)] shows great promise for the vdW-DF method. PMID- 16712104 TI - Creation and destruction of a spin gap in weakly coupled quarter-filled ladders. AB - We investigate weakly coupled quarter-filled ladders with model parameters relevant for NaV(2)O(5) using density-matrix renormalization group calculations on an extended Hubbard model coupled to the lattice. NaV(2)O(5) exhibits super antiferroelectric charge order with a zigzag pattern on each ladder. We show that this order causes a spin dimerization along the ladder and is accompanied by a spin gap of the same magnitude as that observed experimentally. The spin gap is destroyed again at large charge order due to a restructuring of the spins. An analysis of an effective spin model predicts a recreation of the gap by interladder singlets when the charge order increases further. PMID- 16712105 TI - High intrachain hole mobility on molecular wires of ladder-type poly(p phenylenes). AB - We have studied the high-frequency (34 GHz) mobility of positive charge carriers on isolated ladder-type polymer chains in dilute solution. We find that the high frequency mobility is limited by the chain ends on chains as long as 35 monomers. The intrachain motion of charge carriers can be described by one-dimensional diffusion between infinitely high reflecting barriers, representing the chain ends. Our data indicate that the intrachain mobility for ladder-type polymer chains is close to 600 cm(2)/V s. With this high mobility the ladder-type polymer is a promising candidate for future use as an interconnecting wire in molecular electronics. PMID- 16712106 TI - Correlation-induced resonances in transport through coupled quantum dots. AB - We investigate the effect of local electron correlations on transport through parallel quantum dots. The linear conductance as a function of gate voltage is strongly affected by the interplay of the interaction U and quantum interference. We find a pair of novel correlation-induced resonances separated by an energy scale that depends exponentially on U. The effect is robust against a small detuning of the dot energy levels and occurs for arbitrary generic tunnel couplings. It should be observable in experiments on the basis of presently existing double-dot setups. PMID- 16712107 TI - Resonance energy and charge pumping through quantum SINIS contacts. AB - We propose a mechanism of quantum pumping mediated by the spectral flow in a voltage-biased superconductor/insulator/normal-metal/insulator/superconductor quantum junction and realized via the sequential closing of the minigaps in the energy spectrum in resonance with the Josephson frequency. We show that the pumped dc current exhibits giant peaks at rational voltages. PMID- 16712108 TI - Mesoscopic resistance fluctuations in cobalt nanoparticles. AB - We present measurements of mesoscopic resistance fluctuations in cobalt nanoparticles and study how the fluctuations with bias voltage, bias fingerprints, respond to magnetization-reversal processes. Bias fingerprints rearrange when domains are nucleated or annihilated. The domain wall causes an electron wave function-phase shift of approximately equal to 5pi. The phase shift is not caused by the Aharonov-Bohm effect; we explain how it arises from the mistracking effect, where electron spins lag in orientation with respect to the moments inside the domain wall. Dephasing time in Co at 0.03 K is short, tau phi approximately 1 ps, which we attribute to the strong magnetocrystalline anisotropy. PMID- 16712109 TI - Binding of single gold atoms on thin MgO(001) films. AB - In the present Letter the first electron paramagnetic resonance spectra of single metal atoms on a single crystalline oxide surface are presented. For Au atoms on a MgO(001) film investigated here an analysis of the angular dependent resonance positions and the hyperfine coupling to (17)O shows that the atoms are bound on top of oxygen ions on the terrace of the film. This result is in perfect agreement with scanning tunneling microscopy measurements at 5 K presented here. The measured hyperfine matrix components allow an experimental verification of the theoretically proposed binding mechanism of Au atoms on MgO. In particular, the large reduction of the isotropic hyperfine coupling constant of supported Au as compared to free atoms is not due to a charge transfer at the interface but a hybridization of orbitals and a resulting polarization of the unpaired electron. PMID- 16712110 TI - Enhancing the superconducting transition temperature of CeRh 1-x IrxIn5 due to the strong-coupling effects of antiferromagnetic spin fluctuations: an 115In nuclear quadrupole resonance study. AB - We report on systematic evolutions of antiferromagnetic (AFM) spin fluctuations and unconventional superconductivity (SC) in heavy-fermion (HF) compounds CeRh(1 x)Ir(x)In(5) via an (115)In nuclear-quadrupole-resonance experiment. The nuclear spin-lattice relaxation rate 1/T(1) has revealed the marked development of AFM spin fluctuations as approaching an AFM ordered state. Concomitantly, the superconducting transition temperature T(c) and the energy gap Delta0 increase drastically from T(c)= 0.4K and 2Delta0/k(B)T(c)=5 in CeIrIn(5) up to T(c) =1.2K and 2Delta0/k(B)T(c) =8.3 in CeRh(0.3)Ir(0.7)In5 , respectively. The present work suggests that the AFM spin fluctuations in close proximity to the AFM quantum critical point are indeed responsible for the strong-coupling unconventional SC in HF compounds. PMID- 16712111 TI - Critical voltage of a mesoscopic superconductor. AB - We study the influence of a voltage-driven nonequilibrium of quasiparticles on the properties of short mesoscopic superconducting wires. We employ a numerical calculation based upon the Usadel equation. Going beyond linear response, we find a nonthermal energy distribution of the quasiparticles caused by the applied bias voltage. It is demonstrated that this nonequilibrium drives the system from the superconducting state to the normal state, at a current density far below the critical depairing current density. PMID- 16712112 TI - Normal-state diamagnetism of charged bosons in cuprate superconductors. AB - Normal-state orbital diamagnetism of charged bosons quantitatively accounts for recent high-resolution magnetometery results near and above the resistive critical temperature T(c) of superconducting cuprates. The parameter-free descriptions of normal-state diamagnetism, T(c), upper critical fields, and specific heat anomalies support the 3D Bose-Einstein condensation of preformed real-space pairs with a zero off-diagonal order parameter above T(c) at variance with phase fluctuation scenarios of cuprates. PMID- 16712113 TI - Resonating plaquette phase of a quantum six-vertex model. AB - The simplest quantum generalization of the six-vertex model describes fluctuations of the order parameter of the d-density wave (DDW), believed to compete with superconductivity in the high-T(c) superconductors. The ground state of this model undergoes a first order transition from the DDW phase to a resonating plaquette phase as the quantum fluctuations are increased, which is explored with the help of quantum Monte Carlo simulations and analytic considerations involving the n-vector (n = 2) model with cubic anisotropy. In addition to finding a new quantum state, we show that the DDW is robust against a class of quantum fluctuations of its order parameter. The inferred finite temperature phase diagram contains unsuspected multicritical points. PMID- 16712114 TI - Giant magnetocrystalline anisotropies of 4d transition-metal monowires. AB - The magnetocrystalline anisotropy energy (MAE) for ferromagnetic and antiferromagnetic freestanding monowires of 4d transition metals is investigated on the basis of first-principles calculations. Across the 4d series, the easy axis of the magnetization oscillates between two directions: perpendicular and along the wire axis. The largest values of the MAE occur at the end of the series. Giant values of 30-60 meV/atom can be obtained upon stretching Ru or Rh wires. Ru and Pd chains change the magnetization direction upon wire stretching, opening new perspectives in controlling the spin-dependent ballistic conductance in these structures. PMID- 16712115 TI - Imaging spin-reorientation transitions in consecutive atomic Co layers on Ru(0001). AB - By means of spin-polarized low-energy electron microscopy, we show that the magnetic easy axis of one to three atomic-layer thick cobalt films on Ru(0001) changes its orientation twice during deposition: One-monolayer and three monolayer thick films are magnetized in plane, while two-monolayer films are magnetized out of plane. The Curie temperatures of films thicker than one monolayer are well above room temperature. Fully relativistic calculations based on the screened Korringa-Kohn-Rostoker method demonstrate that only for two monolayer cobalt films does the interplay between strain, surface, and interface effects lead to perpendicular magnetization. PMID- 16712116 TI - Tomonaga-Luttinger liquid in a quasi-one-dimensional S = 1 antiferromagnet observed by specific heat measurements. AB - Specific-heat experiments on single crystals of the S = 1 quasi-one-dimensional bond-alternating antiferromagnet Ni(C9H24N4)(NO2)ClO2 (NTENP) have been performed in magnetic fields applied both parallel and perpendicular to the spin chains. We have found for the parallel field configuration that the magnetic specific heat (C(mag)) is proportional to temperature (T) above a critical field H(c), at which the energy gap vanishes, in a temperature region above that of the long-range ordered state. The ratio C(mag)/T increases as the magnetic field approaches H(c) from above. The data are in good quantitative agreement with the prediction of the c= 1 conformal field theory in conjunction with the velocity of the excitations calculated by a numerical diagonalization, providing conclusive evidence for a Tomonaga-Luttinger liquid. PMID- 16712117 TI - Thermally activated depinning of a narrow domain wall from a single defect. AB - We describe the field induced depinning process of a magnetic domain wall (DW) from a single bidimensional nanometric defect. The DW propagates in a wire lithographed on a film with strong perpendicular anisotropy. We observe a statistical distribution of the relaxation time consistent with a Neel-Brown picture of magnetization reversal. This indicates that the nanometric DW can be considered as an ideal monodomain particle switching over a single energy barrier. Such a stochastic character of DW depinning has to be taken into account for spintronic applications. PMID- 16712118 TI - Spin-orbital entanglement and violation of the Goodenough-Kanamori rules. AB - We point out that large composite spin-orbital fluctuations in Mott insulators with t(2g) orbital degeneracy are a manifestation of quantum entanglement of spin and orbital variables. This results in a dynamical nature of the spin superexchange interactions, which fluctuate over positive and negative values, and leads to an apparent violation of the Goodenough-Kanamori rules. PMID- 16712119 TI - Near-surface structural phase transition of SrTiO3 studied with zero-field beta detected nuclear spin relaxation and resonance. AB - We demonstrate that zero-field beta-detected nuclear quadrupole resonance and spin relaxation of low energy (8)Li can be used as a sensitive local probe of structural phase transitions near a surface. We find that the transition near the surface of a SrTiO(3) single crystal occurs at T(c) approximately 150K, i.e., approximately 45K higher than T(c)bulk, and that the tetragonal domains formed below T(c) are randomly oriented. PMID- 16712120 TI - Frustration of tilts and A-site driven ferroelectricity in KNbO3-LiNbO3 alloys. AB - Density functional calculations for K(0.5)Li(0.5)NbO(3) show strong A-site driven ferroelectricity, even though the average tolerance factor is significantly smaller than unity and there is no stereochemically active A-site ion. This is due to the frustration of tilt instabilities by A-site disorder. There are very large off centerings of the Li ions, which contribute strongly to the anisotropy between the tetragonal and rhombohedral ferroelectric states, yielding a tetragonal ground state even without strain coupling. PMID- 16712121 TI - Angle-resolved photoemission spectroscopy of the insulating NaxWO3: Anderson localization, polaron formation, and remnant Fermi surface. AB - The electronic structure of the insulating sodium tungsten bronze, Na(0.025)WO(3), is investigated by high-resolution angle-resolved photoemission spectroscopy. We find that near-E(F) states are localized due to the strong disorder arising from random distribution of Na+ ions in the WO(3) lattice, which makes the system insulating. The temperature dependence of photoemission spectra provides direct evidence for polaron formation. The remnant Fermi surface of the insulator is found to be the replica of the real Fermi surface in the metallic system. PMID- 16712122 TI - Modeling multibody effects in ionic solutions with a concentration dependent dielectric permittivity. AB - We report a new strategy to parametrize effective ion-ion potentials for implicit solvent simulations of charged systems. The effective potential includes a pair term and a Coulomb term that by means of a concentration dependent dielectric permittivity takes into account multibody effects. We demonstrate that this approach allows us to accurately reproduce the solution osmotic properties and the ion coordination up to concentrations of 2.8 M aqueous NaCl. PMID- 16712123 TI - Buckling instability in liquid crystalline physical gels. AB - In a nematic gel we observe a low-energy buckling deformation arising from soft and semisoft elastic modes. We prepare the self-assembled gel by dissolving a coil-side-group liquid-crystalline polymer-coil copolymer in a nematic liquid crystal. The gel has long network strands and a precisely tailored structure, making it ideal for studying nematic rubber elasticity. Under polarized optical microscopy we observe a striped texture that forms when gels uniformly aligned at 35 degrees C are cooled to room temperature. We model the instability using the molecular theory of nematic rubber elasticity, and the theory correctly captures the change in pitch length with sample thickness and polymer concentration. This buckling instability is a clear example of a low-energy deformation that arises in materials where polymer network strains are coupled to the director orientation. PMID- 16712124 TI - Mobility of a one-dimensional confined file of water molecules as a function of file length. AB - Confinement of water by pore geometry to a one-dimensional file of molecules interacting with the pore alters the diffusion coefficient D(W). Here we report an exponential dependence of D(W) on the number of water positions in the pore. The result is based on measurements of single channel water permeabilities of structurally similar peptidic nanopores of different length. The inconsistency with predictions from continuum or kinetic models indicates that pore occupancy is reduced in single file transport. In longer pores (e.g., in aquaporins) the presence of charged residues increases D(W). PMID- 16712125 TI - Synchronization of eukaryotic cells by periodic forcing. AB - We study a cell population described by a minimal mathematical model of the eukaryotic cell cycle subject to periodic forcing that simultaneously perturbs the dynamics of the cell cycle engine and cell growth, and we show that the population can be synchronized in a mode-locked regime. By simplifying the model to two variables, for the phase of cell cycle progression and the mass of the cell, we calculate the Lyapunov exponents to obtain the parameter window for synchronization. We also discuss the effects of intrinsic mitotic fluctuations, asymmetric division, and weak mutual coupling on the pace of synchronization. PMID- 16712126 TI - Designing compliant substrates to regulate the motion of vesicles. AB - By integrating mesoscale models for hydrodynamics and micromechanics, we examine fluid-driven motion of vesicles on compliant surfaces. The vesicles, modeled as fluid-filled elastic shells, represent biological cells or polymeric microcapsules. Focusing on nonspecific interactions between these vesicles and synthetic substrates, we isolate mechanically and topographically patterned surfaces that transmit stop and go instructions, causing the vesicles to halt at specific locations, and with an increase in the flow velocity, to resume moving. For surfaces containing arrays of compliant posts, the substrates also affect the vesicles' gait, causing them to "crawl," "walk," or "jump." The latter behavior could promote the intermixing of reactants that are encapsulated within the microcapsules. Such control over vesicle dynamics can facilitate various biological assays and fabrication of arrays of mobile microreactors. PMID- 16712127 TI - Neural circuitry for recognizing interspike interval sequences. AB - Sensory systems present environmental information to central nervous system as sequences of action potentials or spikes. How do animals recognize these sequences carrying information about their world? We present a biologically inspired neural circuit designed to enable spike pattern recognition. This circuit is capable of training itself on a given interspike interval (ISI) sequence and is then able to respond to presentations of the same sequence. The essential ingredients of the recognition circuit are (a) a tunable time delay circuit, (b) a spike selection unit, and (c) a tuning mechanism using spike timing dependent plasticity of inhibitory synapses. We have investigated this circuit using Hodgkin-Huxley neuron models connected by realistic excitatory and inhibitory synapses. It is robust in the presence of noise represented as jitter in the spike times of the ISI sequence. PMID- 16712128 TI - Systematic series expansions for processes on networks. AB - We use series expansions to study dynamics of equilibrium and nonequilibrium systems on networks. This analytical method enables us to include detailed nonuniversal effects of the network structure. We show that even low order calculations produce results which compare accurately to numerical simulation, while the results can be systematically improved. We show that certain commonly accepted analytical results for the critical point on networks with a broad degree distribution need to be modified in certain cases due to disassortativity; the present method is able to take into account the assortativity at sufficiently high order, while previous results correspond to leading and second order approximations in this method. Finally, we apply this method to real-world data. PMID- 16712129 TI - Transport in weighted networks: partition into superhighways and roads. AB - Transport in weighted networks is dominated by the minimum spanning tree (MST), the tree connecting all nodes with the minimum total weight. We find that the MST can be partitioned into two distinct components, having significantly different transport properties, characterized by centrality--the number of times a node (or link) is used by transport paths. One component, superhighways, is the infinite incipient percolation cluster, for which we find that nodes (or links) with high centrality dominate. For the other component, roads, which includes the remaining nodes, low centrality nodes dominate. We find also that the distribution of the centrality for the infinite incipient percolation cluster satisfies a power law, with an exponent smaller than that for the entire MST. The significance of this finding is that one can improve significantly the global transport by improving a tiny fraction of the network, the superhighways. PMID- 16712130 TI - Comment on "Nature of the polyamorphic transition in ice under pressure". PMID- 16712132 TI - Experimental study of the role of atomic interactions on quantum transport. AB - We report an experimental study of quantum transport for atoms confined in a periodic potential and compare between thermal and Bose-Einstein condensation (BEC) initial conditions. We observe ballistic transport for all values of well depth and initial conditions, and the measured expansion velocity for thermal atoms is in excellent agreement with a single-particle model. For weak wells, the expansion of the BEC is also in excellent agreement with single-particle theory, using an effective temperature. We observe a crossover to a new regime for the BEC case as the well depth is increased, indicating the importance of interactions on quantum transport. PMID- 16712133 TI - Nature of the intrinsic relation between Bloch-band tunneling and modulational instability. AB - In an example of Bose-Einstein condensates embedded in two-dimensional optical lattices, we show that in nonlinear periodic systems modulational instability and interband tunneling are intrinsically related phenomena. By direct numerical simulations we find that tunneling results in attenuation or enhancement of instability. On the other hand, instability results in asymmetric nonlinear tunneling. The effect strongly depends on the band gap structure and it is especially significant in the case of the resonant tunneling. The symmetry of the coherent structures emerging from the instability reflects the symmetry of both the stable and the unstable states between which the tunneling occurs. Our results provide evidence of the profound effect of the band structure on the superfluid-insulator transition. PMID- 16712134 TI - Geometric phase induced by a cyclically evolving squeezed vacuum reservoir. AB - We propose a new way to generate an observable geometric phase by means of a completely incoherent phenomenon. We show how to imprint a geometric phase to a system by adiabatically manipulating the environment with which it interacts. As a specific scheme, we analyze a multilevel atom interacting with a broadband squeezed vacuum bosonic bath. As the squeezing parameters are smoothly changed in time along a closed loop, the ground state of the system acquires a geometric phase. We also propose a scheme to measure such a geometric phase by means of a suitable polarization detection. PMID- 16712135 TI - Trapped fermions with density imbalance in the Bose-Einstein condensate limit. AB - We analyze the effects of imbalancing the populations of two-component trapped fermions, in the Bose-Einstein condensate limit of the attractive interaction between different fermions. Starting from the gap equation with two fermionic chemical potentials, we derive a set of coupled equations that describe composite bosons and excess fermions. We include in these equations the processes leading to the correct dimer-dimer and dimer-fermion scattering lengths. The coupled equations are then solved in the Thomas-Fermi approximation to obtain the density profiles for composite bosons and excess fermions, which are relevant to the recent experiments with trapped fermionic atoms. PMID- 16712136 TI - All bipartite entangled states are useful for information processing. AB - The question of whether all entangled states can be used as a nonclassical resource has remained open so far. Here we provide a conclusive answer to this problem for the case of systems shared by two parties. We show that any entangled state can enhance the teleportation power of some other state. This holds even if the state is bound entangled. PMID- 16712137 TI - Inequalities and separations among assisted capacities of quantum channels. AB - We exhibit quantum channels whose classical and quantum capacities, when assisted by classical feedback, exceed their unassisted classical Holevo capacity. These channels are designed to be noisy in a way that can be corrected with the help of the output and a reference system entangled with part of the input. A similar construction yields quantum channels whose classical capacity, when assisted by two-way classical communication independent of the source, exceeds their classical capacity assisted by feedback alone. We give a hierarchy of capacity inequalities and open questions. PMID- 16712138 TI - Optical implementation of quantum orienteering. AB - We present results from an optical implementation of quantum orienteering, a protocol for communicating directions in space using quantum bits. We show how different types of measurements and encodings can be used to increase the communication efficiency. In particular, if Alice and Bob use two spin- particles for communication and employ joint measurements, they do better than is possible with local operations and classical communication. Furthermore, by using oppositely oriented spins, the achievable communication efficiency is further increased. Finally, we discuss the limitations of an optical approach: our results highlight the usually overlooked nonequivalence of different physical encodings of quantum bits. PMID- 16712140 TI - Spontaneous symmetry breaking of population in a nonadiabatically driven atomic trap: an Ising-class phase transition. AB - We have observed spontaneous symmetry breaking of the population of Brownian particles between two moving potentials in the spatiotemporally symmetric system. Cold atoms preferentially occupy one of the dynamic double-well potentials, produced in the parametrically driven dissipative magneto-optical trap far from equilibrium, above a critical number of atoms. We find that the population asymmetry, which may be interpreted as the biased Brownian motion, can be qualitatively described by the mean-field Ising-class phase transition. This in situ study may be useful for investigation of dynamic phase transition or temporal behavior of critical phenomena. PMID- 16712139 TI - Experimental quantum communication without a shared reference frame. AB - We present an experimental realization of a robust quantum communication scheme [Phys. Rev. Lett. 93, 220501 (2004)] using pairs of photons entangled in polarization and time. Our method overcomes errors due to collective rotation of the polarization modes (e.g., birefringence in optical fiber or misalignment), is insensitive to the phase's fluctuation of the interferometer, and does not require any shared reference frame including time reference, except the need to label different photons. The practical robustness of the scheme is further shown by implementing a variation of the Bennett-Brassard 1984 quantum key distribution protocol over 1 km optical fiber. PMID- 16712141 TI - Diffusionless crystal growth in rapidly solidifying eutectic systems. AB - Using a local nonequilibrium model of solidification, experiments on rapid eutectic growth are analyzed. An analytical solution of a problem of rapid lamellar eutectic growth under local nonequilibrium conditions in the solute diffusion field is found. It is shown that solute diffusion-limited growth of a eutectic pattern is completely finished, and diffusionless growth of the chemically homogeneous crystalline phase begins to proceed at a critical point V = V(D), where V is the solid-liquid interface velocity and V(D) is the solute diffusion speed in bulk liquid. A suppression of eutectic decomposition occurs in the range V > or = V(D) that results in a growth of homogeneous crystal phase with the initial (nominal) chemical composition of the binary system. PMID- 16712142 TI - Indication of a cosmological variation of the proton-electron mass ratio based on laboratory measurement and reanalysis of H2 spectra. AB - Based on highly accurate laboratory measurements of Lyman bands of H2 and an updated representation of the structure of the ground X 1sigma(g)+ and excited B 1sigma(u)+ and C 1pi(u) states, a new set of sensitivity coefficients K(i) is derived for all lines in the H2 spectrum, representing the dependence of their transition wavelengths on a possible variation of the proton-electron mass ratio mu = m(p)/m(e). Included are local perturbation effects between B and C levels and adiabatic corrections. The new wavelengths and K(i) factors are used to compare with a recent set of highly accurate H2 spectral lines observed in the Q 0347-383 and Q 0405-443 quasars, yielding a fractional change in the mass ratio of deltamu/mu = (2.4 +/- 0.6) x 10(-5) for a weighted fit and deltamu/mu = (2.0 +/- 0.6) x 10(-5) for an unweighted fit. This result indicates, at a 3.5sigma confidence level, that mu could have decreased in the past 12 Gyr. PMID- 16712143 TI - Stochastic acceleration in turbulent electric fields generated by 3D reconnection. AB - Electron and proton acceleration in three-dimensional electric and magnetic fields is studied through test particle simulations. The fields are obtained by a three-dimensional magnetohydrodynamic simulation of magnetic reconnection in slab geometry. The nonlinear evolution of the system is characterized by the growth of many unstable modes and the initial current sheet is fragmented with formation of small scale structures. We inject at random points inside the evolving current sheet a Maxwellian distribution of particles. In a relatively short time (less than a millisecond) the particles develop a power-law tail. The acceleration is extremely efficient and the electrons absorb a large percentage of the available energy in a small fraction of the characteristic time of the MHD simulation, suggesting that resistive MHD codes are unable to represent the full extent of particle acceleration. PMID- 16712144 TI - Naturalness in an emergent analogue spacetime. AB - Effective field theories (EFTs) have been widely used as a framework in order to place constraints on the Planck suppressed Lorentz violations predicted by various models of quantum gravity. There are, however, technical problems in the EFT framework when it comes to ensuring that small Lorentz violations remain small--this is the essence of the "naturalness" problem. Herein we present an "emergent" spacetime model, based on the "analogue gravity" program, by investigating a specific condensed-matter system. Specifically, we consider the class of two-component BECs subject to laser-induced transitions between the components, and we show that this model is an example for Lorentz invariance violation due to ultraviolet physics. Furthermore, our model explicitly avoids the naturalness problem, and makes specific suggestions regarding how to construct a physically reasonable quantum gravity phenomenology. PMID- 16712145 TI - Hawking radiation from charged black holes via gauge and gravitational anomalies. AB - Extending the method of Robinson and Wolczek, we show that in order to avoid a breakdown of general covariance and gauge invariance at the quantum level the total flux of charge and energy in each outgoing partial wave of a charged quantum field in a Reissner-Nordstrom black hole background must be equal to that of a (1 + 1)-dimensional blackbody at the Hawking temperature with the appropriate chemical potential. PMID- 16712146 TI - Global universe anisotropy probed by the alignment of structures in the cosmic microwave background. AB - We question the global universe isotropy by probing the alignment of local structures in the cosmic microwave background (CMB) radiation. The original method proposed relies on a steerable wavelet decomposition of the CMB signal on the sphere. The analysis of the first-year Wilkinson Microwave Anisotropy Probe data identifies a mean preferred plane with a normal direction close to the CMB dipole axis, and a mean preferred direction in this plane, very close to the ecliptic poles axis. Previous statistical anisotropy results are thereby synthesized, but further analyses are still required to establish their origin. PMID- 16712148 TI - Lower limit on dark matter production at the CERN Large Hadron Collider. AB - We evaluate the prospects for finding evidence of dark matter production at the CERN Large Hadron Collider. We consider weakly interacting massive particles (WIMPs) and superWIMPs and characterize their properties through model independent parametrizations. The observed relic density then implies lower bounds on dark matter production rates as functions of a few parameters. For WIMPs, the resulting signal is indistinguishable from background. For superWIMPs, however, this analysis implies significant production of metastable charged particles. For natural parameters, these rates may far exceed Drell-Yan cross sections and yield spectacular signals. PMID- 16712147 TI - Improved constraints on D0-D0 mixing in D0 --> K+ pi- decays from the Belle detector. AB - We report the results of a search for D0-D0 mixing in D0 --> K+ pi- decays based on 400 fb(-1) of data accumulated by the Belle detector at KEKB. Both assuming CP conservation and allowing for CP violation, we fit the decay-time distribution for the mixing parameters x and y, as well as for the parameter R(D), the ratio of doubly Cabibbo-suppressed decays to Cabibbo-favored decays. The 95% confidence level region in the (x'2,y') plane is obtained using a frequentist method. Assuming CP conservation, we find x'2 < 0.72 x 10(-3) and -9.9 x 10(-3) < y' < 6.8 x 10(-3) at the 95% confidence level; these are the most stringent constraints on the mixing parameters to date. The no-mixing point (0,0) has a confidence level of 3.9%. Assuming no mixing, we measure R(D) = (0.377 +/- 0.008 +/- 0.005)%. PMID- 16712149 TI - Measurements of the exclusive decays of the upsilon(5S) to meson final states and improved B(s)* mass measurement. AB - Using 420 pb(-1) of data collected on the upsilon(5S) resonance with the CLEO III detector, we reconstruct B mesons in 25 exclusive decay channels to measure or set upper limits on the decay rate of upsilon(5S) into B meson final states. We measure the inclusive B cross section to be sigma(upsilon(5S) --> BB(X)) = (0.177 +/- 0.030 +/- 0.016) nb and make the first measurements of the production rates of sigma(upsilon(5S) --> B*B*) = (0.131 +/- 0.025 +/- 0.014) nb and sigma(upsilon(5S) --> BB*) = (0.043 +/- 0.016 +/- 0.006) nb, respectively. We set 90% confidence level limits of sigma(upsilon(5S) -->BB) < 0.038 nb, sigma(upsilon(5S) --> B(*)B(*)pi) < 0.055 nb and sigma(upsilon(5S) --> BBpipi) < 0.024 nb. We also extract the most precise value of the B(s)* mass to date, M(B(s)*) = (5411.7 +/- 1.6 +/- 0.6) MeV/c2. PMID- 16712152 TI - Experiments with thermalized rare isotope beams from projectile fragmentation: a precision mass measurement of the superallowed beta emitter 38Ca. AB - The mass of the short-lived radio nuclide 38Ca (T(1/2) = 440 ms) has been measured with the 9.4-T Penning trap mass spectrometer of the Low-Energy Beam and Ion Trap Facility. A mass uncertainty of deltam = 280 eV has been achieved, corresponding to deltam/m = 8 x 10(-9). The result makes 38Ca, a superallowed beta emitter, a new candidate to test the conserved-vector-current hypothesis. The experiment is also the first demonstration that short-lived radioactive isotopes produced by projectile fragmentation of relativistic heavy-ion beams can be slowed down and prepared such that precision experiments of this kind are possible. PMID- 16712153 TI - High resolution atomic coherent control via spectral phase manipulation of an optical frequency comb. AB - We demonstrate high resolution coherent control of cold atomic rubidium utilizing spectral phase manipulation of a femtosecond optical frequency comb. Transient coherent accumulation is directly manifested by the enhancement of signal amplitude and spectral resolution via the pulse number. The combination of frequency comb technology and spectral phase manipulation enables coherent control techniques to enter a new regime with natural linewidth resolution. PMID- 16712154 TI - Photodouble ionization dynamics for fixed-in-space H2. AB - The Coulomb explosion of the hydrogen molecule, after absorption of a 76 eV photon, has been studied by momentum imaging the two electrons and the two protons. Absolute fully differential cross sections of high statistical quality are obtained. A subset of the overall data, namely, equal electron-energy sharing, is used to investigate the effects of molecular orientation on the photoelectron angular distribution. Departures from the first-order helium-like model are evident in detection geometries where electron-electron correlation is "frozen." PMID- 16712155 TI - High-resolution photoassociation spectroscopy of ultracold ytterbium atoms by using the intercombination transition. AB - We observed high-resolution photoassociation spectra of laser-cooled ytterbium (Yb) atoms in the spin-forbidden 1S0 - 3P1 intercombination line. The rovibrational levels in the 0u+ state were measured for red detunings of the photoassociation laser ranging from 2.9 MHz to 1.97 GHz with respect to the atomic resonance. The rotational splitting of the vibrational levels near the dissociation limit were fully resolved due to the sub-MHz linewidth of the spectra in contrast to previous measurements using the spin-allowed singlet transition. In addition, from a comparison between the spectra of 174Yb and those of 176Yb, a d-wave shape resonance for 174Yb is strongly suggested. PMID- 16712151 TI - Leptonic and charged kaon decay modes of the phi meson measured in heavy-ion collisions at the CERN super proton synchrotron. AB - We report on results of a measurement of meson production in central Pb-Au collisions at E(lab) = 158A GeV. For the first time in the history of high energy heavy-ion collisions, phi mesons were reconstructed both in the K+K- and the dilepton decay channels in the same experiment. This measurement yields rapidity densities near midrapidity, from the two decay channels, of 2.05 +/- 0.14(stat) +/- 0.25(syst) and 2.04 +/- 0.49(stat) +/- 0.32(syst), respectively. The shape of the measured transverse momentum spectrum is also in close agreement in both decay channels. The data rule out a possible enhancement of the phi yield in the leptonic over the hadronic decay channel of a factor 1.6 or larger at the 95% C.L. This rules out the discrepancy reported in the literature between measurements of the hadronic and dimuon decay channels by two different experiments. PMID- 16712156 TI - Dipole induced transparency in drop-filter cavity-waveguide systems. AB - We show that a waveguide that is normally opaque due to interaction with a drop filter cavity can be made transparent when the drop filter is also coupled to a dipole, even when the vacuum Rabi frequency of the dipole is much less than the cavity decay rate. The condition for transparency is simply achieving large Purcell factors. We describe how this effect can be useful for designing quantum repeaters for long distance quantum communication. PMID- 16712157 TI - Three-dimensional polarization control in microscopy. AB - We propose an approach to optical microscopy that enables full control over the three-dimensional polarization vector at the focal spot of a high-numerical aperture lens. The input field to the lens is linearly polarized and no polarization optics are needed. This technique utilizes the azimuthal spatial degree of freedom of the input field. We find that only a small set of low-order azimuthal spatial harmonics contributes to the focused field on axis, and a simple transformation exists between the linear vector space of these harmonics and the three-dimensional polarization-vector space. Controlling the relative complex weights of these azimuthal harmonics produces any desired three dimensional state of polarization. PMID- 16712158 TI - Inverse design for full control of spontaneous emission using light emitting scattering optical elements. AB - Full control of spontaneous emission is essential in various fields of optics. This work presents an inverse designed light-emitting scattering optical element that includes full control of spontaneously emitted photons (i.e., enhancement at a central frequency and suppression at neighboring frequencies) and directionality of the output beam. This is achieved by embedding a one dimensional optical active element inside a cluster of square shaped gallium arsenide dielectric rods whose positions are optimized by a genetic algorithm. Large spontaneous emission enhancement of > 70 is predicted at the transition wavelength if high-quality sources are employed. Moreover, neighboring wavelengths are simultaneously suppressed over 10 times. Finally, the radiated beam is highly collimated to only 6 degrees and contains 30 times the energy emitted by the source placed in free space. PMID- 16712159 TI - Reaction driven convection around a stably stratified chemical front. AB - A vertical stratification of a light and hot fluid over a heavy and cold one is expected to be stable with regard to buoyancy-driven convection. Here we show that chemical reactions can trigger convection around chemical fronts even in cases where concentration and heat both contribute to a stable density stratification. The balance between intrinsic thermal and solutal density gradients initiated by a spatially localized reaction zone and double diffusive mechanisms are at the origin of a new convective instability, the mechanism of which is explained by a displaced particle argument. Linear stability analysis of a reaction-diffusion-convection model confirmed by nonlinear simulations delimits the instability region in the parameter space spanned by the thermal and solutal Rayleigh numbers. Experimental systems in which to test our theoretical predictions are proposed. PMID- 16712160 TI - Self-propelled Leidenfrost droplets. AB - We report that liquids perform self-propelled motion when they are placed in contact with hot surfaces with asymmetric (ratchetlike) topology. The pumping effect is observed when the liquid is in the Leidenfrost regime (the film-boiling regime), for many liquids and over a wide temperature range. We propose that liquid motion is driven by a viscous force exerted by vapor flow between the solid and the liquid. PMID- 16712162 TI - Real-time measurement of bubbling phenomena in a three-dimensional gas-fluidized bed using ultrafast magnetic resonance imaging. AB - Ultrafast magnetic resonance imaging has been applied for the first time to measure simultaneously both the rise velocities and coalescence of bubbles, and the dynamics of the solid phase in a gas-solid two-phase flow. Here, we consider the hydrodynamics within a gas-fluidized bed of particles of diameter 0.5 mm contained within a column of internal diameter 50 mm; gas velocities in the range of 0.18-0.54 m/s were studied. The data are of sufficient temporal and spatial resolution that bubble size and the evolution of bubble size and velocity following coalescence events are determined. PMID- 16712161 TI - Experimental study of parametric autoresonance in Faraday waves. AB - The excitation of large amplitude nonlinear waves is achieved via parametric autoresonance of Faraday waves. We experimentally demonstrate that phase locking to low amplitude driving can generate persistent high-amplitude growth of nonlinear waves in a dissipative system. The experiments presented are in excellent agreement with theory. PMID- 16712163 TI - Giant bubble pinch-off. AB - Self-similarity has been the paradigmatic picture for the pinch-off of a drop. Here we will show through high-speed imaging and boundary integral simulations that the inverse problem, the pinch-off of an air bubble in water, is not self similar in a strict sense: A disk is quickly pulled through a water surface, leading to a giant, cylindrical void which after collapse creates an upward and a downward jet. Only in the limiting case of large Froude numbers does the purely inertial scaling h(-logh)(1/4) proportional tau(1/2) for the neck radius h [J. M. Gordillo et al., Phys. Rev. Lett. 95, 194501 (2005)] become visible. For any finite Froude number the collapse is slower, and a second length scale, the curvature of the void, comes into play. Both length scales are found to exhibit power-law scaling in time, but with different exponents depending on the Froude number, signaling the nonuniversality of the bubble pinch-off. PMID- 16712164 TI - Transverse characteristics of short-pulse laser-produced ion beams: a study of the acceleration dynamics. AB - We report on first measurements of the transverse characteristics of laser produced energetic ion beams in direct comparison to results for laser accelerated proton beams. The experiments show the same low emittance for ion beams as already found for protons. Additionally, we demonstrate that the divergence is influenced by the charge over mass ratio of the accelerated species. From these observations we deduced scaling laws for the divergence of ions as well as the temporal evolution of the ion source size. PMID- 16712165 TI - Dynamical diffraction theory for wave packet propagation in deformed crystals. AB - We develop a theory for the trajectory of an x ray in the presence of a crystal deformation. A set of equations of motion for an x-ray wave packet including the dynamical diffraction is derived, taking into account the Berry phase as a correction to geometrical optics. The trajectory of the wave packet has a shift of the center position due to a crystal deformation. Remarkably, in the vicinity of the Bragg condition, the shift is enhanced by a factor omega/deltaomega (omega: frequency of an x ray, deltaomega: gap frequency induced by the Bragg reflection). Comparison with the conventional dynamical diffraction theory is also made. PMID- 16712166 TI - Laboratory observation of a nonlinear interaction between shear Alfven waves. AB - An experimental investigation of nonlinear interactions between shear Alfven waves in a laboratory plasma is presented. Two Alfven waves, generated by a resonant cavity, are observed to beat together, driving a pseudomode at the beat frequency. The pseudomode then scatters the Alfven waves, generating a series of sidebands. The observed interaction is very strong, with the normalized amplitude of the driven pseudomode comparable to the normalized magnetic field amplitude (deltaB/B) of the interacting Alfven waves. PMID- 16712167 TI - Synthesis of novel transition metal nitrides IrN2 and OsN2. AB - Two new transition metal nitrides, IrN2 and OsN2, were synthesized at high pressures and temperatures using laser-heated diamond-anvil cell techniques. Synchrotron x-ray diffraction was used to determine the structures of novel nitrides and the equations of states of both the parent metals as well as the newly synthesized materials. The compounds have bulk moduli comparable with those of the traditional superhard materials. For IrN2, the measured bulk modulus [K0 = 428(12) GPa] is second only to that of diamond (K0 = 440 GPa). Ab initio calculations indicate that both compounds have a metal:nitrogen stoichiometry of 1:2 and that nitrogen intercalates in the lattice of the parent metal in the form of single-bonded N-N units. PMID- 16712168 TI - Dynamical identification of a threshold instability in Si-doped heterofullerenes. AB - We rationalize the origins of a threshold instability and the mechanism of finite temperature fragmentation in highly Si-doped C(60-m)Si(m) heterofullerenes via a first-principles approach. Cage disruption is driven by enhanced fluctuations of the most internal Si atoms. These are located within fully segregated Si regions neighboring the C-populated part of the cage. The predominance of inner Si atoms over those involved in Si-C bonds marks the transition from thermally stable to unstable C(60-m)Si(m) systems at m = 20. PMID- 16712169 TI - Theory of the phonon Hall effect in paramagnetic dielectrics. AB - Based upon Raman spin-lattice interaction, we propose a theoretical model for the phonon Hall effect in paramagnetic dielectrics, which was discovered recently in an experiment [C. Strohm, G. L. J. A. Rikken, and P. Wyder, Phys. Rev. Lett. 95, 155901 (2005).]. The phonon Hall effect is revealed to be a phonon analogue to the anomalous Hall effect in electron systems. The thermal Hall conductivity is calculated by using the Kubo formula. Our theory reproduces the essential experimental features of the phonon Hall effect, including the sign, magnitude, and linear magnetic field dependence of the thermal Hall conductivity. PMID- 16712170 TI - Cascade of shocks in inertial liquid-liquid dewetting. AB - We study the inertial dewetting of water films (A) (thickness e) deposited on highly hydrophobic liquid substrates (B). On these ideal surfaces, thin films can be made which dewet at large velocities obeying under those conditions the Culick law for the bursting of soap films. The rim collecting the water film can become coupled to the surface waves characterized by a surface tension gamma(B) upstream of the rim (coated substrate) and gamma = gamma(B) downstream, where the water film has dried. Upon decreasing the thickness, we observe a sequence of two hydraulic shocks during the dewetting inducing gravity waves behind the rim, and capillary waves ahead. PMID- 16712171 TI - Selective analysis of molecular states by functionalized scanning tunneling microscopy tips. AB - Selective analysis of molecular states in scanning tunneling microscopy (STM) has so far been achieved in a few cases by tuning the bias range of the STM in high resolution measurements. Correspondingly, perylene adsorbed in a close-packed monolayer on Ag(110) is imaged mainly through the pi states of the molecule. By contrast, functionalizing the STM tip with a perylene molecule leads to a mismatch between the energy levels of the STM tip and the molecule adsorbates and, instead, images only the metal states of the underlying silver surface. The observation opens a route for better energy selectivity in electron transport measurements through organic interfaces. PMID- 16712150 TI - Top-quark mass measurement from dilepton events at CDF II. AB - We report a measurement of the top-quark mass using events collected by the CDF II detector from pp collisions at square root of s = 1.96 TeV at the Fermilab Tevatron. We calculate a likelihood function for the top-quark mass in events that are consistent with tt --> bl(-)nu(l)bl'+ nu'(l) decays. The likelihood is formed as the convolution of the leading-order matrix element and detector resolution functions. The joint likelihood is the product of likelihoods for each of 33 events collected in 340 pb(-1) of integrated luminosity, yielding a top quark mass M(t) = 165.2 +/- 6.1(stat) +/- 3.4(syst) GeV/c2. This first application of a matrix-element technique to tt --> bl+ nu(l)bl'- nu(l') decays gives the most precise single measurement of M(t) in dilepton events. Combined with other CDF run II measurements using dilepton events, we measure M(t) = 167.9 +/- 5.2(stat) +/- 3.7(syst) GeV/c2. PMID- 16712172 TI - Dielectric fluctuations and the origins of noncontact friction. AB - Dielectric fluctuations underlie a wide variety of physical phenomena, from ion mobility in electrolyte solutions and decoherence in quantum systems to dynamics in glass-forming materials and conformational changes in proteins. Here we show that dielectric fluctuations also lead to noncontact friction. Using high sensitivity, custom fabricated, single crystal silicon cantilevers we measure energy losses over poly(methyl methacrylate), poly(vinyl acetate), and polystyrene thin films. A new theoretical analysis, relating noncontact friction to the dielectric response of the film, is consistent with our experimental observations. This work constitutes the first direct, mechanical detection of noncontact friction due to dielectric fluctuations. PMID- 16712173 TI - Metastable structures and recombination pathways for atomic hydrogen on the graphite (0001) surface. AB - We present scanning tunneling microscopy results which reveal the existence of two distinct hydrogen dimer states on graphite basal planes. Density functional theory calculations allow us to identify the atomic structure of these states and to determine their recombination and desorption pathways. Direct recombination is only possible from one of the two dimer states. This results in increased stability of one dimer species and explains the puzzling double peak structure observed in temperature programmed desorption spectra for hydrogen on graphite. PMID- 16712174 TI - Disentanglement time of polymers determines the onset of rim instabilities in dewetting. AB - Molecular relaxations determine the viscoelastic properties of polymers, which, in turn, control macroscopic processes like dewetting. Here, we demonstrate experimentally that the onset of rim instabilities is correlated with the longest relaxation ("reptation") time of the dewetting polymer. Conversely, such experiments allow us to determine the reptation time of polystyrene in thin films as a function of molecular weight. Our approach opens up new possibilities for testing rheological properties of polymers confined in thin films. PMID- 16712175 TI - Conformational molecular switch of the azobenzene molecule: a scanning tunneling microscopy study. AB - We propose to utilize azobenzene as a nanomolecular switch which can be triggered by transmitting electrons above threshold biases. The effect is explained by an electron impact trans-cis conformational change of the isolated azobenzene molecules. The molecular electronic states of both isomers have been measured with spatially resolved scanning tunneling microscopy or spectroscopy, leading to suggested transition pathways of the electron-induced isomerization. PMID- 16712176 TI - Quasicrystalline electronic states of a one-dimensionally modulated Ag film. AB - Ag films on GaAs(110) exhibit a one-dimensional quasiperiodic modulation, resulting in a Fibonacci sequence of parallel stripes with two different widths. Valence level photoemission shows that the Ag electronic states acquire a unique character along the quasiperiodic direction, distinctively manifested by a hierarchy of energy level replicas and avoided crossings at characteristic intervals in reciprocal space. These observations demonstrate the strong influence of the one-dimensional quasiperiodic potential on the Ag film states. PMID- 16712177 TI - Phonon effects on spin-charge separation in one dimension. AB - Phonon effects on spin-charge separation in one dimension are investigated through the calculation of one-electron spectral functions in terms of the recently developed cluster perturbation theory together with an optimized phonon approach. It is found that the retardation effect due to the finiteness of phonon frequency suppresses the spin-charge separation and eventually makes it invisible in the spectral function. By comparing our results with experimental data of TTF TCNQ, it is observed that the electron-phonon interaction must be taken into account when interpreting the angle-resolved photoemission spectroscopy data. PMID- 16712178 TI - Metal to insulator transition in films of molecularly linked gold nanoparticles. AB - We report a metal to insulator transition (MIT) in disordered films of molecularly linked gold nanoparticles (NPs). As the number of carbons (n) of alkanedithiol linker molecules (C(n)S2) is varied, resistance (R) at low temperature (T = 2 K) and at 200 K, as well as trends in R vs T data at intermediate temperatures, all point to an MIT occurring at n = 5. We describe these results in a context of a Mott-Hubbard MIT. We find that all insulating samples (n > or = 5) exhibit a universal scaling behavior R approximately exp[(T0/T)nu] with nu = 0.65, and all metallic samples (n < or = 5) exhibit weaker R-T dependencies than bulk gold. We discuss these observations in terms of competitive thermally activated processes and strong, T-independent elastic scattering, respectively. PMID- 16712179 TI - Specific heat of delta-Pu stabilized by Am. AB - Detailed specific heat C(p) measurements of delta-Pu stabilized by Am (8%-20%) were performed in the temperature range 4.5-300 K. The coefficient of the electronic specific heat gamma, which reflects the quasiparticle density of states at the Fermi level E(F), is smaller than originally assumed and, depending on the estimate of phonon contributions, a value between 35 and 55 mJ/mol K2 can be deduced for Pu-8% Am. For higher Am concentrations, which expand the lattice, gamma decreases slightly with the Am content. An applied magnetic field of 9 T had no effect on C(p). The results strongly suggest that itinerant 5f states at E(F) are not appropriate for describing delta-Pu. PMID- 16712180 TI - Electrically tunable resistance of a metal. AB - Electric field-induced tuning of material properties is usually restricted to nonmetals such as semiconductors and piezoelectric ceramics. We show that variations of the electrical resistance of a metal (Pt) in the range of several percent can be reversibly induced at low charging voltages making use of a nanocrystallite-electrolyte composite. The charge-induced resistance variation is analyzed taking into account the modification of the charge carrier density and scattering rate by surface charging. The contribution arising from the charge induced variation of the lattice constant is found to be small. PMID- 16712181 TI - Spontaneous spin polarization in quantum point contacts. AB - We use spatial spin separation by a magnetic focusing technique to probe the polarization of quantum point contacts. The point contacts are fabricated from p type GaAs/AlGaAs heterostructures. A finite polarization is measured in the low density regime, when the conductance of a point contact is tuned to < 2e2/h. Polarization is stronger in samples with a well-defined "0.7 structure." PMID- 16712182 TI - Electronic effects in the length distribution of atom chains. AB - Gold deposited on Si(553) leads to self-assembly of atomic chains, which are broken into finite segments by defects. Scanning tunneling microscopy is used to investigate the distribution of chain lengths and the correlation between defects separating the chains. The length distribution reveals oscillations that indicate changes in the cohesive energy as a function of chain length. We present a possible interpretation in terms of the electronic scattering vectors at the Fermi surface of the surface states. The pairwise correlation function between defects shows long-range correlations that extend beyond nearest-neighbor defects, indicating coupling between chains. PMID- 16712183 TI - Multipeak Kondo effect in one- and two-electron quantum dots. AB - We have fabricated a few-electron quantum dot that can be tuned down to zero electrons while maintaining strong coupling to the leads. Using a nearby quantum point contact as a charge sensor, we can determine the absolute number of electrons in the quantum dot. We find several sharp peaks in the differential conductance, occurring at both zero and finite source-drain bias, for the one- and two-electron quantum dot. We attribute the peaks at finite bias to a Kondo effect through excited states of the quantum dot and investigate the magnetic field dependence of these Kondo resonances. PMID- 16712184 TI - Evidence for temperature-dependent electron band dispersion in pentacene. AB - Evidence for temperature-dependent electron band dispersion in a pentacene thin film polymorph on graphite is provided by angle- and energy-dependent ultraviolet photoelectron spectroscopy. The bands derived from the highest occupied molecular orbital exhibit dispersion of approximately 190 meV at room temperature, and approximately 240 meV at 120 K. Intermolecular electronic coupling in pentacene thin films is thus confirmed to be dependent on temperature and possibly crystal structure, as suggested by additional infrared absorption measurements. PMID- 16712185 TI - Mesoscopic fluctuations of nonlinear conductance of chaotic quantum dots. AB - The nonlinear dc conductance of a two-terminal chaotic cavity is investigated. The fluctuations of the conductance (anti)symmetric with respect to magnetic-flux inversion through multichannel cavities are found analytically for arbitrary temperature, magnetic field, and interaction strength. For few-channel dots the effect of dephasing is investigated numerically. A comparison with recent experimental data is provided. PMID- 16712186 TI - High-energy spin excitations in the electron-doped superconductor Pr(0.88)LaCe(0.12)CuO(4-delta) with T(c) = 21 K. AB - We use high-resolution inelastic neutron scattering to study the low-temperature magnetic excitations of the electron-doping superconductor Pr(0.88)LaCe(0.12)CuO(4-delta) (T(c) = 21 +/- 1 K) over a wide energy range (4 meV < or = homega < or = 330 meV). The effect of electron doping is to cause a wave vector (Q) broadening in the low-energy (homega < or = 80 meV) commensurate spin fluctuations at (0.5, 0.5) and to suppress the intensity of spin-wave-like excitations at high energies (homega > or = 100 meV). This leads to a substantial redistribution in the spectrum of the local dynamical spin susceptibility chi''(omega), and reveals a new energy scale similar to that of the lightly hole doped YB2Cu3O(6.353) (T(c) = 18 K). PMID- 16712187 TI - Superconducting phase coherent electron transport in proximity conical ferromagnets. AB - We report superconducting phase-periodic conductance oscillations in ferromagnetic wires with interfaces to conventional superconductors. The ferromagnetic wires were made of Ho, a conical ferromagnet. The distance between the interfaces was much larger than the singlet superconducting penetration depth. We explain the observed oscillations as due to the long-range penetration of an unusual helical triplet component of the order parameter that is generated at the superconductor/ferromagnet interfaces and maintained by the intrinsic rotating magnetization of Ho. PMID- 16712188 TI - Doping dependence of the coupling of electrons to bosonic modes in the single layer high-temperature Bi2Sr2CuO6 superconductor. AB - A recent highlight in the study of high-T(c) superconductors is the observation of band renormalization or self-energy effects on the quasiparticles. This is seen in the form of kinks in the quasiparticle dispersions as measured by photoemission and interpreted as signatures of collective bosonic modes coupling to the electrons. Here we compare for the first time the self-energies in an optimally doped and strongly overdoped, nonsuperconducting single-layer Bi cuprate (Bi2Sr2CuO6). In addition to the appearance of a strong overall weakening, we also find that the weight of the self-energy in the overdoped system shifts to higher energies. We present evidence that this is related to a change in the coupling to c-axis phonons due to the rapid change of the c-axis screening in this doping range. PMID- 16712189 TI - Localized and delocalized excitons: resonant inelastic x-ray scattering in La(2 x)Sr(x)NiO4 and La(2-x)Sr(x)CuO4. AB - The dynamics of doped charge in an antiferromagnetic lattice is central to the description of the insulator-metal transition that occurs on doping the parent high T(c) compounds. In this work we use high resolution resonant inelastic x-ray scattering to investigate the dynamics of the charge-transfer exciton by measuring its energy dispersion in two prototype compounds, La2CuO4 and La2NiO4. We show that this behavior is radically different in the cuprate with respect to a system known to exhibit strong polaronic behavior, namely, the nickelate: the exciton is mobile in the cuprate while it is well localized in the nickelate. Using a simple Wannier-Mott model we can estimate the total hole plus electron effective mass in the cuprate to be 3.5 +/- 0.3 m(e) which would exclude strong localization in the undoped cuprate. PMID- 16712190 TI - Stabilizing superconductivity in nanowires by coupling to dissipative environments. AB - We present a theory for a finite-length superconducting nanowire coupled to an environment. We show that in the absence of dissipation quantum phase slips always destroy superconductivity, even at zero temperature. Dissipation stabilizes the superconducting phase. We apply this theory to explain the "antiproximity effect" recently seen by Tian et al. in zinc nanowires. PMID- 16712191 TI - Theory of electron mediated Mn-Mn interactions in quantum dots. AB - We present a theory of interaction of magnetic Mn ions depending strongly on the number (Ne) of electrons in a quantum dot. For closed electronic shells, we derive the RKKY interaction and its dependence on magnetic ion positions, quantum dot energy quantization omega0, and the number of filled shells Ns. For partially filled shells, the many-electron magnetopolaron effect leads to effective carrier mediated ferromagnetic Mn-Mn interactions. The dependence of the magnetopolaron energy on magnetic ion positions, quantum dot energy quantization omega0, and the number of electrons Ne is predicted. PMID- 16712192 TI - Colossal magnetocapacitance and colossal magnetoresistance in HgCr2S4. AB - We present a detailed study of the dielectric and charge transport properties of the antiferromagnetic cubic spinel HgCr2S4. Similar to the findings in ferromagnetic CdCr2S4, the dielectric constant of HgCr2S4 becomes strongly enhanced in the region below 60-80 K, which can be ascribed to polar relaxational dynamics triggered by the onset of ferromagnetic correlations. In addition, the observation of polarization hysteresis curves indicates the development of ferroelectric order below about 70 K. Moreover, our investigations in external magnetic fields up to 5 T reveal the simultaneous occurrence of magnetocapacitance and magnetoresistance of truly colossal magnitudes in this material. PMID- 16712193 TI - Dispersion of surface plasmon polaritons on metal wires in the terahertz frequency range. AB - We report the experimental and theoretical study of the dispersive behavior of surface plasmon polaritons (SPPs) on cylindrical metal surfaces in the terahertz frequency range. Time-domain measurements of terahertz SPPs propagating on metal wires reveal a unique structure that is inconsistent with a simple extrapolation of the high frequency portion of the dispersion diagram for SPPs on a planar metal surface, and also distinct from that of SPPs on metal nanowires observed at visible and near-infrared frequencies. The results are consistent with a numerical solution of Maxwell's equations, showing that the dispersive behavior of SPPs on a cylindrical metal surface at terahertz frequencies is quite different from that of SPPs on a flat surface. These findings indicate the increasing importance of skin effects for SPPs in the terahertz range, as well as the enhancement of such effects on curved surfaces. PMID- 16712194 TI - Enhanced Raman scattering from individual semiconductor nanocones and nanowires. AB - We report strong enhancement (approximately 10(3)) of the spontaneous Raman scattering from individual silicon nanowires and nanocones as compared with bulk Si. The observed enhancement is diameter (d), excitation wavelength (lambda(laser)), and incident polarization state dependent, and is explained in terms of a resonant behavior involving incident electromagnetic radiation and the structural dielectric cross section. The variation of the Raman enhancement with d, lambda(laser), and polarization is shown to be in good agreement with model calculations of scattering from an infinite dielectric cylinder. PMID- 16712195 TI - Electronic origins of structural distortions in post-transition metal oxides: experimental and theoretical evidence for a revision of the lone pair model. AB - Structural distortions in post-transition metal oxides are often explained in terms of the influence of sp hybrid "lone pairs." Evidence is presented here showing that this model must be revised. The electronic structures of prototypically distorted alpha-PbO and alpha-Bi2O3 have been measured by high resolution x-ray photoemission and soft x-ray emission spectroscopies. In contrast with the expectations of the lone pair model, a high density of metal 6s states is observed at the bottom of the valence band. The measurements are consistent with the results of density functional theory calculations. PMID- 16712196 TI - Origin of fine structure in si photoelectron spectra at silicon surfaces and interfaces. AB - Using a first-principles approach, we investigate the origin of the fine structure in Si 2p photoelectron spectra at the Si(100)-(2 x 1) surface and at the Si(100)-SiO2 interface. Calculated and measured shifts show very good agreement for both systems. By using maximally localized Wannier functions, we clearly identify the shifts resulting from the electronegativity of second neighbor atoms. The other shifts are then found to be proportional to the average bond-length variation around the Si atom. Hence, in combination with accurate modeling, photoelectron spectroscopy can provide a direct measure of the strain field at the atomic scale. PMID- 16712197 TI - In-plane and out-of-plane ferroelectric instabilities in epitaxial SrTiO3 films. AB - The in-plane and out-of-plane ferroelectric instabilities in compressed (100) epitaxial SrTiO3 films were examined by infrared reflection spectroscopy. The strongly stiffened in-plane soft mode frequency softened very slowly on cooling. On the other hand, the silent mode appeared at around 150 K, indicating an out-of plane ferroelectric transition. This behavior points to a split of in-plane and out-of-plane ferroelectric instability temperatures due to the lowered symmetry of the SrTiO3 lattice caused by mechanical misfit strain. Infrared spectroscopy provides a possibility to detect such an effect in the strained epitaxial ferroelectric films. PMID- 16712198 TI - Escape configuration lattice near the nematic-isotropic transition: tilt analogue of blue phases. AB - We predict the possible existence of a new phase of liquid crystals near the nematic-isotropic transition. This phase is an achiral, tilt analogue of the blue phase and is composed of a lattice of double-tilt, escape-configuration cylinders. We discuss the structure and the stability of this phase and provide an estimate of the lattice parameter. PMID- 16712199 TI - Dynamics of impact cratering in shallow sand layers. AB - When a solid sphere impacts a shallow layer of sand deposited on a solid surface, a crater can be obtained. The dynamics of the opening of the crater can be followed accurately. During this opening, the radius of the crater can be conveniently modeled by an exponential saturation with a well-defined time constant. The crater then closes up partially once the opening phase is over as the sand avalanches down the slope of the crater. We here present a detailed study of the full dynamics of the crater formation as well as the dynamics of the corrola formed during this process. A simple model accounts for most of our observations. PMID- 16712200 TI - Mesoscopic theory of critical fluctuations in isolated granular gases. AB - Fluctuating hydrodynamics is used to describe the total energy fluctuations of a freely evolving gas of inelastic hard spheres near the threshold of the clustering instability. They are shown to be governed only by vorticity fluctuations that also lead to a renormalization of the average total energy. The theory predicts a power-law divergent behavior of the scaled second moment of the fluctuations, and a scaling property of their probability distribution, both in agreement with simulations results. A more quantitative comparison between theory and simulation for the critical amplitudes and the form of the scaling function is also carried out. PMID- 16712201 TI - Pumping fluids with periodically beating grafted elastic filaments. AB - Using Brownian dynamics simulations, we investigate the pumping efficiency of an array of periodically beating semiflexible filaments that are grafted to a surface. Full hydrodynamic interactions among and within filaments and no slip at the surface are considered. Optimal pumping is obtained for a characteristic ratio of applied forward-backward torques and filament persistence length. For independently driven filaments, phase locking between neighboring filaments occurs autonomously via hydrodynamic coupling, giving rise to significantly enhanced pumping efficiency. PMID- 16712202 TI - Comment on "Reconciling coulomb dissociation and radiative capture measurements". PMID- 16712204 TI - Comment on "Negative refraction at optical frequencies in nonmagnetic two component molecular media". PMID- 16712206 TI - A theory of electromagnetic fluctuations for metallic surfaces and van der waals interactions between metallic bodies. AB - A new general expression is derived for the fluctuating electromagnetic field outside a metal surface in terms of its surface impedance. It provides a generalization to real metals of Lifshitz theory of molecular interactions between dielectric solids. The theory is used to compute the radiative heat transfer between two parallel metal surfaces at different temperatures. It is shown that a measurement of this quantity may provide an experimental resolution of a long-standing controversy about the effect of thermal corrections on the Casimir force between real metal plates. PMID- 16712207 TI - Critical temperature and thermodynamics of attractive fermions at unitarity. AB - The unitarity regime of the BCS-BEC crossover can be realized by diluting a system of two-component lattice fermions with an on-site attractive interaction. We perform a systematic-error-free finite-temperature simulation of this system by diagrammatic determinant Monte Carlo method. The critical temperature in units of Fermi energy is found to be T(C)/epsilonF=0.152(7). We also report the behavior of the thermodynamic functions, and discuss the issues of thermometry of ultracold Fermi gases. PMID- 16712208 TI - High-order quantum resonances observed in a periodically kicked Bose-Einstein condensate. AB - We have observed high-order quantum resonances in a realization of the quantum delta-kicked rotor, using Bose-condensed Na atoms subjected to a pulsed standing wave of laser light. These resonances occur for pulse intervals that are rational fractions of the Talbot time, and are characterized by ballistic momentum transfer to the atoms. The condensate's narrow momentum distribution not only permits the observation of the quantum resonances at 3/4 and 1/3 of the Talbot time, but also allows us to study scaling laws for the resonance width in quasimomentum and pulse interval. PMID- 16712209 TI - Observation of bunching of two bell states. AB - The bunching of two single photons on a beam splitter is a fundamental quantum effect, first observed by Hong, Ou, and Mandel. It is a unique interference effect that relies only on the photons' indistinguishability and not on their relative phase. We generalize this effect by demonstrating the bunching of two Bell states, created in two passes of a nonlinear crystal, each composed of two photons. When the two Bell states are indistinguishable, phase-insensitive destructive interference prevents the outcome of fourfold coincidence between the four spatial-polarization modes. For certain combinations of the two Bell states, we demonstrate the opposite effect of antibunching. We relate this result to the number of distinguishable modes in parametric down-conversion. PMID- 16712210 TI - Formation of supermassive black holes through fragmentation of torodial supermassive stars. AB - We investigate new paths to supermassive black hole formation by considering the general relativistic evolution of a differentially rotating polytrope with a toroidal shape. We find that this polytrope is unstable to nonaxisymmetric modes, which leads to a fragmentation into self-gravitating, collapsing components. In the case of one such fragment, we apply a simplified adaptive mesh refinement technique to follow the evolution to the formation of an apparent horizon centered on the fragment. This is the first study of the onset of nonaxisymmetric dynamical instabilities of supermassive stars in full general relativity. PMID- 16712211 TI - Brane gas-driven bulk expansion as a precursor stage to brane inflation. AB - We propose a new way of obtaining slow-roll inflation in the context of higher dimensional models motivated by string and M theory. In our model, all extra spatial dimensions are orbifolded. The initial conditions are taken to be a hot dense bulk brane gas which drives an initial phase of isotropic bulk expansion. This phase ends when a weak potential between the orbifold fixed planes begins to dominate. For a wide class of potentials, a period during which the bulk dimensions decrease sufficiently slowly to lead to slow-roll inflation of the three dimensions parallel to the orbifold fixed planes will result. Once the separation between the orbifold fixed planes becomes of the string scale, a repulsive potential due to string effects takes over and leads to a stabilization of the radion modes. The conversion of bulk branes into radiation during the phase of bulk contraction leads to reheating. PMID- 16712212 TI - Production of axions by cosmic magnetic helicity. AB - We investigate the effects of an external magnetic helicity production on the evolution of the cosmic axion field. It is shown that a helicity larger than (few x 10(-15) G)2 Mpc, if produced at temperatures above a few GeV, is in contradiction with the existence of the axion, since it would produce too much of an axion relic abundance. PMID- 16712213 TI - Moduli space of non-abelian vortices. AB - We completely determine the moduli space M(N,k) of k vortices in U(N) gauge theory with N Higgs fields in the fundamental representation. Its open subset for separated vortices is found as the symmetric product (CxCP(N-1))k/(see text)k. Orbifold singularities of this space correspond to coincident vortices and are resolved resulting in a smooth moduli manifold. The relation to Kahler quotient construction is discussed. PMID- 16712214 TI - Percolation effects in very-high-energy cosmic rays. AB - Cosmic ray data at high energies present a number of well-known puzzles. At very high energies (E approximately 10(20) eV) there are indications of a discrepancy between ground array experiments and fluorescence detectors. On the other hand, the dependence of the depth of the shower maximum Xmax with the primary energy shows a change in slope (E approximately 10(17) eV) which is usually explained assuming a composition change. Both effects could be accounted for in models predicting that above a certain energy showers would develop deeper in the atmosphere. In this Letter we argue that this can be done naturally by including percolation effects in the description of the shower development, which cause a change in the behavior of the inelasticity K above E approximately 10(17) eV. PMID- 16712219 TI - 15C-15F Charge symmetry and the 14C(n,gamma)15C reaction puzzle. AB - The low-energy reaction 14C(n,gamma)15C provides a rare opportunity to test indirect methods for the determination of neutron capture cross sections by radioactive isotopes versus direct measurements. It is also important for various astrophysical scenarios. Currently, puzzling disagreements exist between the 14C(n,gamma)15C cross sections measured directly, determined indirectly, and calculated theoretically. To solve this puzzle, we offer a strong test based on a novel idea that the amplitudes for the virtual 15C-->14C + n and the real 15F - >14O + p decays are related. Our study of this relation, performed in a microscopic model, shows that existing direct and some indirect measurements strongly contradict charge symmetry in the 15C and 15F mirror pair. This brings into question the experimental determinations of the astrophysically important (n,gamma) cross sections for short-lived radioactive targets. PMID- 16712218 TI - First measurement of the rho spectral function in high-energy nuclear collisions. AB - We report on a precision measurement of low-mass muon pairs in 158 AGeV indium indium collisions at the CERN SPS. A significant excess of pairs is observed above the yield expected from neutral meson decays. The unprecedented sample size of 360,000 dimuons and the good mass resolution of about 2% allow us to isolate the excess by subtraction of the decay sources. The shape of the resulting mass spectrum is consistent with a dominant contribution from pi+pi- -->rho -->mu+mu- annihilation. The associated space-time averaged spectral function shows a strong broadening, but essentially no shift in mass. This may rule out theoretical models linking hadron masses directly to the chiral condensate. PMID- 16712221 TI - Experimental constraints on polarizability corrections to hydrogen hyperfine structure. AB - We present a state-of-the-art evaluation of the polarizability corrections--the inelastic nucleon corrections--to the hydrogen ground-state hyperfine splitting using analytic fits to the most recent data. We find a value Delta(pol)=1.3+/-0.3 ppm. This is 1-2 ppm smaller than the value of Delta(pol) deduced using hyperfine splitting data and elastic nucleon corrections obtained from modern form factor fits. PMID- 16712220 TI - Deep sub-barrier fusion enhancement in the 6He + 206Pb reaction. AB - The fusion of 6He with 206Pb has been studied at energies close to and below the Coulomb barrier. The experiment was carried out at the Dubna Radioactive Ion Beams complex of FLNR, JINR. The 6He beam intensity was about 5 x 10(6) pps, the maximum energy being 60.3+/-0.4 MeV. The yield of the 210Po isotope, produced in the 2n-evaporation channel, demonstrates an extremely large enhancement of the sub-barrier fusion cross section as compared with the 4He+208Pb reaction. This enhancement is most likely due to the mechanism of "sequential fusion" with an intermediate neutron transfer from 6He to the Pb nucleus with positive Q values. PMID- 16712215 TI - Observation of a near-threshold enhancement in the omega(phi) mass spectrum from the doubly OZI-suppressed decay J/psi-->gamma(omega)phi. AB - An enhancement near threshold is observed in the omega(phi) invariant mass spectrum from the doubly Okubo-Zweig-Iizuka-suppressed decays of J/psi- >gamma(omega)phi, based on a sample of 5.8 x 10(7) J/psi events collected with the BESII detector. A partial wave analysis shows that this enhancement favors JP=0+, and its mass and width are M=1812(+19)(-26)(stat)+/-18(syst) MeV/c2 and Gamma=105+/-20(stat)+/-28(syst) MeV/c2. The product branching fraction is determined to be B(J/psi-->gammaX)B(X-->omega(phi))=[2.61+/-0.27(stat)+/ 0.65(syst)]x10(-4). PMID- 16712222 TI - Rapid proton transfer mediated by a strong laser field. AB - Kinetic energy distributions of ejected from a polyatomic molecule, anthraquinone, subjected to 60 fs, 800 nm laser pulses of intensity between 0.2 and 4.0 x 10(14) W x cm(-2), reveal field-driven restructuring of the molecule prior to Coulomb explosion. Calculations demonstrate fast intramolecular proton migration into a field-dressed metastable potential energy minimum. The proton migration occurs in the direction perpendicular to the polarization of the laser field. Rapid field-mediated isomerization is an important new phenomenon in coupling of polyatomic molecules with intense lasers. PMID- 16712217 TI - Double-hadron leptoproduction in the nuclear medium. AB - The first measurements of double-hadron production in deep-inelastic scattering within the nuclear medium were made with the HERMES spectrometer at DESY HERA using a 27.6 GeV positron beam. By comparing data for deuterium, nitrogen, krypton, and xenon nuclei, the influence of the nuclear medium on the ratio of double-hadron to single-hadron yields was investigated. Nuclear effects on the additional hadron are clearly observed, but with little or no difference among nitrogen, krypton, or xenon, and with smaller magnitude than effects seen on previously measured single-hadron multiplicities. The data are compared with models based on partonic energy loss or prehadronic scattering and with a model based on a purely absorptive treatment of the final-state interactions. Thus, the double-hadron ratio provides an additional tool for studying modifications of hadronization in nuclear matter. PMID- 16712216 TI - Charmonium Decays of Y(4260), psi(4160), and psi(4040). AB - Using data collected with the CLEO detector operating at the CESR e+e- collider at sqrt[s]=3.97-4.26 GeV, we investigate 15 charmonium decay modes of the psi(4040), psi(4160), and Y(4260) resonances. We confirm, at 11 sigma significance, the BABAR Y(4260)-->pi+pi- J/psi discovery, make the first observation of Y(4260)--> pi(0)pi(0) J/psi (5.1 sigma), and find the first evidence for Y(4260)-->K+K- J/psi(3.7 sigma). We measure e+e- cross sections at sqrt[s]=4.26 GeV as sigma(pi+pi- J/psi)=58(+12)(-10)+/-4 pb, sigma(pi(0)pi(0) J/psi)=23(+12)(-8)+/-1 pb, and sigma(K+K- J/psi)=9(+9)(-5)+/-1 pb, in which the uncertainties are statistical and systematic, respectively. Upper limits are placed on other decay rates from all three resonances. PMID- 16712223 TI - Relativistic coupled-cluster theory of atomic parity nonconservation: application to 137Ba+. AB - We report the result of our ab initio calculation of the 6s2S1/2-->5d2D3/2 parity nonconserving electric dipole transition amplitude in 137Ba+ based on relativistic coupled-cluster theory. Considering single, double, and partial triple excitations, we have achieved an accuracy of less than 1%. If the accuracy of our calculation can be matched by the proposed parity nonconservation experiment in Ba+ for the above transition, then the combination of the two results would provide an independent nonaccelerator test of the standard model of particle physics. PMID- 16712224 TI - New mass value for 7Li. AB - A high-accuracy mass measurement of 7Li was performed with the SMILETRAP Penning trap mass spectrometer via a cyclotron frequency comparison of 7Li3+ and H2+. A new atomic-mass value of 7Li has been determined to be 7.016 003 425 6(45) u with a relative uncertainty of 0.63 ppb. It has uncovered a discrepancy as large as 14sigma (1.1 microu) deviation relative to the literature value given in the Atomic-Mass Evaluation AME 2003. The importance of the improved and revised 7Li mass value, for calibration purposes in nuclear-charge radii and atomic-mass measurements of the neutron halos 9Li and 11Li, is discussed. PMID- 16712225 TI - Lost topological (Berry) phase factor in electronic structure calculations. Example: the ozone molecule. AB - It is shown that standard computations of electronic structures of polyatomic systems that yield the global minimum configuration and vibrational frequencies may be faulty if the symmetry of this configuration is lower than the highest possible one and the origin of this distortion, which is always due to the Jahn Teller effect, is neglected; this may lead, in particular, to the loss of the Berry phase factor that changes the vibronic energy level spectrum and which we show to be present even when there are no apparent conical intersections. The general case and the ozone molecule are analyzed. PMID- 16712226 TI - One-dimensional Bose chemistry: effects of nonintegrability. AB - Three-body collisions of ultracold identical Bose atoms under tight cylindrical confinement are analyzed. A Feshbach resonance in two-body collisions is described by a two-channel zero-range interaction. Elimination of the closed channel in the three-body problem reduces the interaction to a one-channel zero range one with an energy-dependent strength. The related integrable Lieb-Liniger McGuire (LLMG) model, with an energy-independent strength, forbids all chemical processes, such as three-atom association and diatom dissociation, as well as reflection in atom-diatom collisions. The resonant case is analyzed by a numerical solution of the Faddeev equations. The results demonstrate that as the internal symmetry of the LLMG model is lifted, reflection and chemical reactions become allowed and may be observed in experiments. PMID- 16712227 TI - Violation of the exponential-decay law at long times. AB - First-principles quantum mechanical calculations show that the exponential-decay law for any metastable state is only an approximation and predict an asymptotically algebraic contribution to the decay for sufficiently long times. In this Letter, we measure the luminescence decays of many dissolved organic materials after pulsed laser excitation over more than 20 lifetimes and obtain the first experimental proof of the turnover into the nonexponential decay regime. As theoretically expected, the strength of the nonexponential contributions scales with the energetic width of the excited state density distribution whereas the slope indicates the broadening mechanism. PMID- 16712228 TI - Universal optical amplification without nonlinearity. AB - We propose and experimentally realize a new scheme for universal phase insensitive optical amplification. The presented scheme relies only on linear optics and homodyne detection, thus circumventing the need for nonlinear interaction between a pump field and the signal field. The amplifier demonstrates near optimal quantum noise limited performance for a wide range of amplification factors. PMID- 16712229 TI - Quantum lithography with classical light. AB - We show how to achieve subwavelength diffraction and imaging with classical light, previously thought to require quantum fields. By correlating wave vector and frequency in a narrow band, multiphoton detection process that uses Doppleron type resonances, we show how to achieve arbitrary focal and image plane patterning with classical laser light at submultiples of the Rayleigh limit, with high efficiency, visibility, and spatial coherence. A frequency-selective measurement process thus allows one to simulate, semiclassically, the path-number correlations that distinguish a quantum entangled field. PMID- 16712230 TI - Spectral phase distribution retrieval through coherent control of harmonic generation. AB - The temporal intensity distribution of the third harmonic of a Ti:sapphire laser generated in Xe gas is fully reconstructed from its spectral phase and amplitude distributions. The spectral phases are retrieved by cross correlating the fundamental laser frequency field with that of the third harmonic, in a three laser versus one harmonic photon coupling scheme. The third harmonic spectral amplitude distribution is extracted from its field autocorrelation. The measured pulse duration is found to be in agreement with that expected from lowest order perturbation theory both for unstretched and chirped pulses. PMID- 16712232 TI - Mode softening, ferroelectric transition, and tunable photonic band structures in a point-dipole crystal. AB - We study the photonic band structure of cubic crystals of point dipoles. It is shown that in contrast to earlier claims these systems cannot have an omnidirectional photonic band gap. For sufficiently large plasma frequencies, however, they exhibit softening of photonic bands, leading to (anti)ferroelectric ordering of the dipoles and the possibility to open and tune directional band gaps by external electric fields. The model studied may be realized through lattices of quantum dots. PMID- 16712231 TI - Correlation transfer and diffusion of ultrasound-modulated multiply scattered light. AB - We develop a temporal correlation transfer equation (CTE) and a temporal correlation diffusion equation (CDE) for ultrasound-modulated multiply scattered light. These equations can be applied to an optically scattering medium with embedded optically scattering and absorbing objects to calculate the power spectrum of light modulated by a nonuniform ultrasound field. We present an analytical solution based on the CDE and Monte Carlo simulation results for light modulated by a cylinder of ultrasound in an optically scattering slab. We further validate with experimental measurements the numerical calculations for an actual ultrasound field. The CTE and CDE are valid for moderate ultrasound pressures and on a length scale comparable with the optical transport mean-free path. These equations should be applicable to a wide spectrum of conditions for ultrasound modulated optical tomography of soft biological tissues. PMID- 16712233 TI - Localized polaritons and second-harmonic generation in a resonant medium with quadratic nonlinearity. AB - We derive model equations for the propagation of ultrashort pulses in materials with resonant linear and quadratic nonlinear responses and find approximate soliton solutions describing all-bright and dark-bright polaritons. We report the specific phase matching condition for efficient 2nd harmonic generation, which involves detuning from the resonance. We also demonstrate that the 2nd harmonic emission by the polaritonic pulses can lead to reduction of their group velocity, having zero as a theoretical limit. Our analytical results are supported by numerical simulations. PMID- 16712234 TI - Optical spin-to-orbital angular momentum conversion in inhomogeneous anisotropic media. AB - We demonstrate experimentally an optical process in which the spin angular momentum carried by a circularly polarized light beam is converted into orbital angular momentum, leading to the generation of helical modes with a wave-front helicity controlled by the input polarization. This phenomenon requires the interaction of light with matter that is both optically inhomogeneous and anisotropic. The underlying physics is also associated with the so-called Pancharatnam-Berry geometrical phases involved in any inhomogeneous transformation of the optical polarization. PMID- 16712236 TI - Dynamical weights and enhanced synchronization in adaptive complex networks. AB - Dynamical organization of connection weights is studied in scale-free networks of chaotic oscillators, where the coupling strength of a node from its neighbors develops adaptively according to the local synchronization property between the node and its neighbors. We find that when complete synchronization is achieved, the coupling strength becomes weighted and correlated with the topology due to a hierarchical transition to synchronization in heterogeneous networks. Importantly, such an adaptive process enhances significantly the synchronizability of the networks, which could have meaningful implications in the manipulation of dynamical networks. PMID- 16712235 TI - Quantum accelerator modes from the Farey tree. AB - We show that mode locking finds a purely quantum nondissipative counterpart in atom-optical quantum accelerator modes. These modes are formed by exposing cold atoms to periodic kicks in the direction of the gravitational field. They are anchored to generalized Arnol'd tongues, parameter regions where driven nonlinear classical systems exhibit mode locking. A hierarchy for the rational numbers known as the Farey tree provides an ordering of the Arnol'd tongues and hence of experimentally observed accelerator modes. PMID- 16712237 TI - Lorentz force velocimetry. AB - We describe a noncontact technique for velocity measurement in electrically conducting fluids. The technique, which we term Lorentz force velocimetry (LFV), is based on exposing the fluid to a magnetic field and measuring the drag force acting upon the magnetic field lines. Two series of measurements are reported, one in which the force is determined through the angular velocity of a rotary magnet system and one in which the force on a fixed magnet system is measured directly. Both experiments confirm that the measured signal is a linear function of the flow velocity. We then derive the scaling law that relates the force on a localized distribution of magnetized material to the velocity of an electrically conducting fluid. This law shows that LFV, if properly designed, has a wide range of potential applications in metallurgy, semiconductor crystal growth, and glass manufacturing. PMID- 16712238 TI - Single-point velocity statistics of forced and decaying two-dimensional turbulence. AB - The single-point (SP) velocity statistics are investigated in forced and decaying two-dimensional turbulence in a flowing soap film. It is shown that the probability distribution functions (PDF) in both cases deviate from a Gaussian distribution, which is normally anticipated in turbulent fluid flows. In the forced turbulence case, the tail of the SP velocity PDF decays faster than Gaussian (termed the sub-Gaussian) and can be correlated with the forcing statistics on small scales. In the decaying-turbulence case, the SP velocity PDF evolves from a sub-Gaussian to a super-Gaussian behavior as a function of time. However, in all times, the locally averaged vorticity remains normally distributed. While our forced turbulence data may be explained by a recent theory proposed by Falkovich et al., the decaying-turbulence data remain unexplained. PMID- 16712239 TI - Tailored terahertz pulses from a laser-modulated electron beam. AB - We present a new method to generate steady and tunable, coherent, broadband terahertz radiation from a relativistic electron beam modulated by a femtosecond laser. We have demonstrated this in the electron storage ring at the Advanced Light Source. Interaction of an electron beam with a femtosecond laser pulse copropagating through a wiggler modulates the electron energies within a short slice of the electron bunch with about the same duration of the laser pulse. The bunch develops a longitudinal density perturbation due to the dispersion of electron trajectories, and the resulting hole emits short pulses of temporally and spatially coherent terahertz pulses synchronized to the laser. We present measurements of the intensity and spectra of these pulses. This technique allows tremendous flexibility in shaping the terahertz pulse by appropriate modulation of the laser pulse. PMID- 16712240 TI - Ultrafast dynamics of strongly coupled plasmas. AB - The ultrafast dynamics of a strongly coupled plasma following an energy landscape shift is studied theoretically and with simulation. To lowest order in time, the inertial dynamics on the new landscape can be characterized by the plasma microfield, which, for the randomly ordered case of an ultracold neutral plasma, is dominated by nearest neighbor interactions. Formation of the pair correlation function arises after ballistic overshoot, which leads to oscillations in the effective temperature. Warm dense matter systems are also considered in this context. PMID- 16712241 TI - Nonlinear theory for relativistic plasma wakefields in the blowout regime. AB - We present a theory for nonlinear, multidimensional plasma waves with phase velocities near the speed of light. It is appropriate for describing plasma waves excited when all electrons are expelled out from a finite region by either the space charge of a short electron beam or the radiation pressure of a short intense laser. It works very well for the first bucket before phase mixing occurs. We separate the plasma response into a cavity or blowout region void of all electrons and a sheath of electrons just beyond the cavity. This simple model permits the derivation of a single equation for the boundary of the cavity. It works particularly well for narrow electron bunches and for short lasers with spot sizes matched to the radius of the cavity. It is also used to describe the structure of both the accelerating and focusing fields in the wake. PMID- 16712243 TI - Bose-Einstein condensation of incommensurate solid 4He. AB - It is pointed out that the simulation computation of energy performed so far cannot be used to decide if the ground state of solid 4He has the number of lattice sites equal to the number of atoms (commensurate state) or if it is different (incommensurate state). The best variational wave function, a shadow wave function, gives an incommensurate state, but the equilibrium concentration of vacancies remains to be determined. We have computed the one-body density matrix in solid 4He for the incommensurate state by means of an exact ground state projector method in which incommensurability occurs spontaneously. We find a vacancy induced Bose-Einstein condensation of about 0.23 atoms per vacancy at a pressure of 54 bar. This means that bulk solid 4He is supersolid at low enough temperature if the exact ground state is incommensurate. PMID- 16712242 TI - Observation of a fast electron beam emitted along the surface of a target irradiated by intense femtosecond laser pulses. AB - A novel fast electron beam emitting along the surface of a target irradiated by intense laser pulses is observed. The beam is found to appear only when the plasma density scale length is small. Numerical simulations reveal that the electron beam is formed due to the confinement of the surface quasistatic electromagnetic fields. The results are of interest for potential applications of fast electron beams and deep understanding of the cone-target physics in the fast ignition related experiments. PMID- 16712244 TI - Axial-strain-induced torsion in single-walled carbon nanotubes. AB - Using classical molecular dynamics and empirical potentials, we show that the axial deformation of single-walled carbon nanotubes is coupled to their torsion. The axial-strain-induced torsion is limited to chiral nanotubes-graphite sheets rolled around an axis that breaks its symmetry. Small strain behavior is consistent with chirality and curvature-induced elastic anisotropy (CCIEA)-carbon nanotube rotation is equal and opposite in tension and compression, and decreases with curvature and chirality. The large-strain compressive response is remarkably different. The coupling progressively decreases, in contrast to the tensile case, and changes its sign at a critical compressive strain. Thereafter, it untwists with increasing axial strain and then rotates in the opposite direction, i.e., the same sense as under tension. This suggests that the response is now dictated by a combination of nonlinear elasticity and CCIEA. PMID- 16712245 TI - Local structure of a rolled-up single crystal: an X-ray microdiffraction study of individual semiconductor nanotubes. AB - Crystals with cylindrical symmetry, not existing in nature, are mimicked by the roll-up of single-crystalline and highly strained semiconductor bilayers. Exploiting this, the local structure of such individual rolled-up nanotubes is locally probed and quantified nondestructively by x-ray microbeam diffraction. A comparison to simulations, based on the minimization of the elastic energy, allows us to determine layer thicknesses and lattice parameter distributions within the strongly curved bilayers. PMID- 16712246 TI - Debye screening of dislocations. AB - Debye-like screening by edge dislocations of some externally given stress is studied by means of a variational approach to coarse grained field theory. Explicitly given are the force field and the induced geometrically necessary dislocation (GND) distribution, in the special case of a single glide axis in 2D, for (i) a single edge dislocation and (ii) a dislocation wall. Numerical simulation demonstrates that the correlation in relaxed dislocation configurations is in good agreement with the induced GND in case (i). Furthermore, the result (ii) well predicts the experimentally observed decay length for the GND developing close to grain boundaries. PMID- 16712247 TI - Localization transition of the three-dimensional lorentz model and continuum percolation. AB - The localization transition and the critical properties of the Lorentz model in three dimensions are investigated by computer simulations. We give a coherent and quantitative explanation of the dynamics in terms of continuum percolation theory and obtain an excellent matching of the critical density and exponents. Within a dynamic scaling ansatz incorporating two divergent length scales we achieve data collapse for the mean-square displacements and identify the leading corrections to scaling. We provide evidence for a divergent non-Gaussian parameter close to the transition. PMID- 16712249 TI - Two-dimensional roughening of adsorbate islands in thermodynamic equilibrium. AB - Equilibrium fluctuations of islands of adsorbed O atoms on Ru(0001) were investigated by scanning tunneling microscopy (STM), density functional theory calculations (DFT) and Monte Carlo (MC) simulations. Very ramified (2 x 2)-O islands were observed by high-speed STM that point to complex interactions between the O atoms. The DFT calculations show that, in addition to pairwise attractive interactions between third-nearest neighbors, a repulsive three-body interaction exists between these. MC simulations that include three-body interactions reproduce the observed ordering behavior. PMID- 16712248 TI - Laser-induced microexplosion confined in the bulk of a sapphire crystal: evidence of multimegabar pressures. AB - Extremely high pressures (approximately 10 TPa) and temperatures (5 x 10(5) K) have been produced using a single laser pulse (100 nJ, 800 nm, 200 fs) focused inside a sapphire crystal. The laser pulse creates an intensity over 10(14) W/cm2 converting material within the absorbing volume of approximately 0.2 microm3 into plasma in a few fs. A pressure of approximately 10 TPa, far exceeding the strength of any material, is created generating strong shock and rarefaction waves. This results in the formation of a nanovoid surrounded by a shell of shock affected material inside undamaged crystal. Analysis of the size of the void and the shock-affected zone versus the deposited energy shows that the experimental results can be understood on the basis of conservation laws and be modeled by plasma hydrodynamics. Matter subjected to record heating and cooling rates of 10(18) K/s can, thus, be studied in a well-controlled laboratory environment. PMID- 16712250 TI - Capillary condensation in atomic scale friction: how water acts like a glue. AB - We present atomic-scale friction force measurements that strongly suggest that the capillary condensation of water between a tungsten tip and a graphite surface leads to the formation of ice at room temperature. This phenomenon increases the friction force, introduces a short-term memory in the form of an elastic response against shearing, and allows us to "write" a temporary line of ice on a hydrophobic surface. Rearrangements of the condensate are shown to take place on a surprisingly slow time scale of seconds. PMID- 16712251 TI - Effect of step stiffness and diffusion anisotropy on the meandering of a growing vicinal surface. AB - We study the step meandering instability on a surface characterized by the alternation of terraces with different properties, as in the case of Si(001). The interplay between diffusion anisotropy and step stiffness induces a finite wavelength instability corresponding to a meandering mode. The instability sets in beyond a threshold value which depends on the relative magnitudes of the destabilizing flux and the stabilizing stiffness difference. The meander dynamics is governed by the conserved Kuramoto-Sivashinsky equation, which display spatiotemporal coarsening. PMID- 16712252 TI - Pressure-induced metal-insulator transition in LaMnO3 is not of Mott-Hubbard type. AB - Calculations employing the local density approximation combined with static and dynamical mean field theories (LDA+U and LDA+DMFT) indicate that the metal insulator transition observed at 32 GPa in paramagnetic LaMnO3 at room temperature is not a Mott-Hubbard transition, but is caused by orbital splitting of the majority-spin eg bands. For LaMnO3 to be insulating at pressures below 32 GPa, both on-site Coulomb repulsion and Jahn-Teller distortion are needed. PMID- 16712253 TI - Angle-dependent magnetoresistance in the weakly incoherent interlayer transport regime in a layered organic conductor. AB - We present comparative studies of the orientation effect of a strong magnetic field on the interlayer resistance of alpha-(BEDT-TTF)2KHg(SCN)4 samples characterized by different crystal quality. We find striking differences in their behavior, which is attributed to the breakdown of the coherent charge transport across the layers in the lower quality sample. In the latter case, the nonoscillating magnetoresistance background is essentially a function of only the out-of-plane field component, in contradiction to the existing Fermi-liquid theories. PMID- 16712254 TI - Static dielectric properties of carbon nanotubes from first principles. AB - We characterize the response of isolated single-wall (SWNT) and multiwall (MWNT) carbon nanotubes and nanotube bundles to static electric fields using first principles calculations and density-functional theory. The longitudinal polarizability of SWNTs scales as the inverse square of the band gap, while in MWNTs and bundles it is given by the sum of the polarizabilities of the constituent tubes. The transverse polarizability of SWNTs is insensitive to band gaps and chiralities and is proportional to the square of the effective radius; in MWNTs, the outer layers dominate the response. The transverse response is intermediate between metallic and insulating, and a simple electrostatic model based on a scale-invariance relation captures accurately the first-principles results. The dielectric response of nonchiral SWNTs in both directions remains linear up to very high values of applied field. PMID- 16712255 TI - Adiabatic pumping in interacting systems. AB - A dc current can be pumped through an interacting system by periodically varying two independent parameters such as the magnetic field and a gate potential. We present a general expression for the adiabatic pumping current in interacting systems, written in terms of instantaneous properties of the system at equilibrium, and find the limits of its applicability. This expression generalizes the scattering approach for noninteracting particles. We apply our formula for a quantum critical system that exhibits the two-channel Kondo effect, where single particle excitations are not well defined. We find that if the quantum critical point is contained in the pumping trajectory, the pumped spin between the channels approaches h, and if it is not contained in the trajectory, the spin approaches zero when the temperature T --> 0. We discuss the non-Fermi liquid features of this system at finite T. PMID- 16712256 TI - Molecular transport junctions: current from electronic excitations in the leads. AB - Using a model comprising a two-level bridge connecting free electron reservoirs we show that coupling of a molecular bridge to electron-hole excitations in the leads can markedly effect the source-drain current through a molecular junction. In some cases, e.g., molecules that exhibit strong charge transfer transitions, the contribution from electron-hole excitations can exceed the Landauer elastic current and dominate the observed conduction. PMID- 16712258 TI - Surface segregation and backscattering in doped silicon nanowires. AB - By means of ab initio simulations, we investigate the structural, electronic, and transport properties of boron and phosphorus doped silicon nanowires. We find that impurities always segregate at the surface of unpassivated wires, reducing dramatically the conductance of the surface states. Upon passivation, we show that for wires as large as a few nanometers in diameter, a large proportion of dopants will be trapped and electrically neutralized at surface dangling bond defects, significantly reducing the density of carriers. Important differences between p- and n-type doping are observed. Our results rationalize several experimental observations. PMID- 16712257 TI - Nonlinear spin current and magnetoresistance of molecular tunnel junctions. AB - We report on a theoretical study of spin-polarized quantum transport through a Ni bezenedithiol(BDT)-Ni molecular magnetic tunnel junction (MTJ). Our study is based on carrying out density functional theory within the Keldysh nonequilibrium Green's function formalism, so that microscopic details of the molecular MTJ are taken into account from first principles. A magnetoresistance ratio of approximately 27% is found for the Ni-BDT-Ni MTJ which declines toward zero as bias voltage is increased. The spin currents are nonlinear functions of bias voltage, even changing sign at certain voltages due to specific features of the coupling between molecular states and magnetic leads. PMID- 16712259 TI - Mesoscopic versus macroscopic division of current fluctuations. AB - We investigate the current shot noise at a three terminal node in which one of the branches contains a noise generating source and the correlations are measured between the currents flowing through the other two branches. Interestingly, if the node is macroscopic, the current correlations are positive, whereas for a quantum coherent mesoscopic node antibunching of electrons leads to negative correlations. We present specific predictions which permit the experimental investigation of the crossover from the quantum mechanical noise division to the macroscopic noise division. PMID- 16712260 TI - Bloch inductance in small-capacitance Josephson junctions. AB - We show that the electrical impedance of a small-capacitance Josephson junction also includes, in addition to the capacitive term -i/(omega)CB, an inductive term i(omega)LB. Similar to the known Bloch capacitance CB(q), the Bloch inductance LB(q) also depends periodically on the quasicharge, q, and its maximum value achieved at q=e(mod 2e) always exceeds the value of the Josephson inductance of this junction LJ(phi) at fixed phi=0. The effect of the Bloch inductance on the dynamics of a single junction and a one-dimensional array is described. PMID- 16712261 TI - Spontaneous symmetry-breaking vortex lattice transitions in pure niobium. AB - We report an extensive investigation of magnetic vortex lattice (VL) structures in single crystals of pure niobium with the magnetic field applied parallel to a fourfold symmetry axis, so as to induce frustration between the cubic crystal symmetry and hexagonal VL coordination expected in an isotropic situation. We observe new VL structures and phase transitions; all the VL phases observed (including those with an exactly square unit cell) spontaneously break some crystal symmetry. One phase even has the lowest possible symmetry of a two dimensional Bravais lattice. This is quite unlike the situation in high-Tc or borocarbide superconductors, where VL structures orient along particular directions of high crystal symmetry. The causes of this behavior are discussed. PMID- 16712262 TI - Large anisotropic normal-state magnetoresistance in clean MgB2 thin films. AB - We report a large normal-state magnetoresistance with temperature-dependent anisotropy in very clean epitaxial MgB2 thin films (residual resistivity much smaller than 1 microOmega cm) grown by hybrid physical-chemical vapor deposition. The magnetoresistance shows a complex dependence on the orientation of the applied magnetic field, with a large magnetoresistance (Delta(rho)/(rho)0=136%) observed for the field H perpendicular ab plane. The angular dependence changes dramatically as the temperature is increased, and at high temperatures the magnetoresistance maximum changes to H||ab. We attribute the large magnetoresistance and the evolution of its angular dependence with temperature to the multiple bands with different Fermi surface topology in MgB2 and the relative scattering rates of the sigma and pi bands, which vary with temperature due to stronger electron-phonon coupling for the sigma bands. PMID- 16712263 TI - Supercurrent-induced temperature gradient across a nonequilibrium SNS Josephson junction. AB - Using tunneling spectroscopy, we have measured the local electron energy distribution function in the normal part of a superconductor-normal metal superconductor (SNS) Josephson junction containing an extra lead to a normal reservoir. In the presence of simultaneous supercurrent and injected quasiparticle current, the distribution function exhibits a sharp feature at very low energy. The feature is odd in energy and odd under reversal of either the supercurrent or the quasiparticle current direction. The feature represents an effective temperature gradient across the SNS Josephson junction that is controllable by the supercurrent. PMID- 16712264 TI - Index theoretic characterization of d-wave superconductors in the vortex state. AB - We study analytically the low energy spectrum of a lattice d-wave superconductor in the vortex lattice state. For an inversion symmetric hc/2e vortex lattice and in the presence of particle-hole symmetry we prove an index theorem that imposes a lower bound on the number of zero-energy modes. Generic cases are constructed in which this bound exceeds the number of zero modes of an equivalent lattice of hc/e vortices, despite the identical point group symmetries. The quasiparticle spectrum around the zero modes is doubly degenerate and exhibits a Dirac-like dispersion, with velocities that become universal functions of Delta(0)/t in the limit of low magnetic field. For weak particle-hole symmetry breaking, the gapped state can be characterized by a topological quantum number, related to spin-Hall conductivity, which generally differs in the cases of the hc/2e and hc/e vortex lattices. PMID- 16712265 TI - Observation of a Griffiths-like phase in the magnetocaloric compound Tb5Si2Ge2. AB - The onset of a Griffiths-like phase has been observed in Tb5Si2Ge2 (TC=110 K) by means of magnetic susceptibility and small-angle neutron scattering experiments. We show the growth of a ferromagnetic cluster system characterized by an inverse susceptibility exponent lower than unity at TC rho(+) rho(-) decays with the Belle detector at the KEKB e(+) e(-) collider using 253 Fb(-1) of data. We obtain B = [22.8 +/- 3.8(stat)(+2.3)(-2.6)(syst)] x 10(-6), f(L) = 0.941 (+0.034)(-0.040)(stat) +/- 0.030(syst). A = 0.00 +/- 0.30(stat) +/- 0.09(syst) and S = 0.08 +/- 0.09(syst). These values are used to constrain the Cabibbo Kobayashi-Maskawa phase ; the solution consistent with the standard model is phi(2) = (88 +/- 17) degrees or 59 degrees < phi(2) < 115 degrees at 90% C.L. PMID- 16712293 TI - Coherent control of ultracold molecule dynamics in a magneto-optical trap by use of chirped femtosecond laser pulses. AB - We have studied the effects of chirped femtosecond laser pulses on the formation of ultracold molecules in a Rb magneto-optical trap. We have found that application of chirped femtosecond pulses suppressed the formation of (85)Rb and (87)Rb(2) a(3)sigma(+)(u) molecules in contrast to comparable nonchirped pulses, cw illumination, and background formation rates. Variation of the amount of chirp indicated that this suppression is coherent in nature, suggesting that coherent control is likely to be useful for manipulating the dynamics of ultracold quantum molecular gases. PMID- 16712294 TI - Kinematically complete study of dissociative ionization of by ion impact. AB - We present a kinematically complete study of dissociative ionization of D(2) by 13.6 MeV/u S(15+) ions. The experiment allows us to unravel the competing mechanisms, namely, direct single ionization, autoionization of doubly excited states, ionization excitation, and double ionization, and to analyze the corresponding electron angular distribution from fixed-in-space molecules. The conclusions are supported by theoretical calculations in which the correlated motion of all electrons and nuclei and the interferences between them are described from first principles. PMID- 16712295 TI - Interference swapping in scattering from a nonlocal quantum target. AB - We describe a new and distinctive interferometry in which a probe particle scatters off a superposition of locations of a single free target particle. Probe particles scattering off a single free "mirror" (in one dimension) or a single free "slit" (in two dimensions) can "swap" interference with the superposed target states. The condition for interference is loss of orthogonality of the target states and reduces, in simple examples, to transfer of orthogonality from target to probe states. We analyze experimental parameters and conditions necessary for interference to be observed. PMID- 16712296 TI - High finesse opto-mechanical cavity with a movable thirty-micron-size mirror. AB - We report on the demonstration of a high finesse micro-optomechanical system and identify potential applications ranging from optical cooling to weak force detection to massive quantum superpositions. The system consists of a high quality diameter flat dielectric mirror cut from a larger substrate with a focused ion beam and attached to an atomic force microscope cantilever. Cavity ring-down measurements performed on a 25 mm long Fabry-Perot cavity with the 30 microm mirror at one end show an optical finesse of 2100. Numerical calculations show that the finesse is not diffraction limited and that orders of magnitude higher finesse should be possible. A mechanical quality factor of more than 10(5) at pressures below 10(-3) mbar is demonstrated for the cantilever with a mirror attached. PMID- 16712297 TI - Experimental demonstration of self-collimation inside a three-dimensional photonic crystal. AB - We present our experimental demonstration of self-collimation inside a three dimensional (3D) simple cubic photonic crystal at microwave frequencies. The photonic crystal was designed with unique dispersion property and fabricated by a high precision computer-controlled machine. The self-collimation modes were excited by a grounded waveguide feeding and detected by a scanning monopole. Self collimation of electromagnetic waves in the 3D photonic crystal was demonstrated by measuring the 3D field distribution, which was shown as a narrow collimated beam inside the 3D photonic crystal but a diverged beam in the absence of the photonic crystal. PMID- 16712298 TI - From finite to infinite volumes: removal of boundaries in diffuse wave imaging. AB - In this Letter, we present a method that removes the contribution of the boundaries on the measurements from highly scattering media, transforming the signals captured from a bounded medium to measurements that would have been obtained if no boundary were present. This approach opens new possibilities in tomographic imaging in diffuse media as it eliminates the need for explicitly modeling boundaries and significantly simplifies reconstruction requirements. PMID- 16712300 TI - Edge of chaos in a parallel shear flow. AB - We study the transition between laminar and turbulent states in a Galerkin representation of a parallel shear flow, where a stable laminar flow and a transient turbulent flow state coexist. The regions of initial conditions where the lifetimes show strong fluctuations and a sensitive dependence on initial conditions are separated from the ones with a smooth variation of lifetimes by an object in phase space which we call the "edge of chaos." We describe techniques to identify and follow the edge, and our results indicate that the edge is a surface. For low Reynolds numbers we find that the surface coincides with the stable manifold of a periodic orbit, whereas at higher Reynolds numbers it is the stable set of a higher-dimensional chaotic object. PMID- 16712299 TI - Antenna model for wire lasers. AB - An antenna model is proposed for long (L >> A) lasers with subwavelength cross sections (wire lasers). It is shown that the far-field pattern of the wire lasers is determined by the ratio of the wavelength to the length. The radiation of the wire laser is predicted to be concentrated in a narrow beam theta approximately radical(2lambda/L) for laser modes where the longitudinal phase velocity is in synchronism with the velocity of light in air. Experimental results obtained using a terahertz quantum cascade wire laser are in agreement with the model. PMID- 16712301 TI - Dissimilar electro-osmotic flow and ionic current recirculation patterns in porous media detected by NMR mapping experiments. AB - Random-site percolation clusters were milled into ceramic (polar) and polystyrene (nonpolar) plates as a paradigm for porous media or complex microsystem channel networks. The pore space was filled with electrolyte solutions. Using NMR microscopy techniques, maps of the following quantities were recorded: (i) flow velocity driven by external pressure gradient, (ii) electro-osmotic flow (EOF) velocity, (iii) ionic current density in the presence of EOF, (iv) ionic current density in the absence of EOF. As far as possible, the experiments were supplemented by computational fluid dynamics simulations. It is shown that electro-osmotic flow as well as the electric current density include vortices and recirculation patterns. Remarkably, all transport patterns turned out to be dissimilar, and the occurrence and positions of vortices do not coincide in the different maps. PMID- 16712302 TI - Polygons on a rotating fluid surface. AB - We report a novel and spectacular instability of a fluid surface in a rotating system. In a flow driven by rotating the bottom plate of a partially filled, stationary cylindrical container, the shape of the free surface can spontaneously break the axial symmetry and assume the form of a polygon rotating rigidly with a speed different from that of the plate. With water, we have observed polygons with up to 6 corners. It has been known for many years that such flows are prone to symmetry breaking, but apparently the polygonal surface shapes have never been observed. The creation of rotating internal waves in a similar setup was observed for much lower rotation rates, where the free surface remains essentially flat [J. M. Lopez, J. Fluid Mech. 502, 99 (2004). We speculate that the instability is caused by the strong azimuthal shear due to the stationary walls and that it is triggered by minute wobbling of the rotating plate. PMID- 16712304 TI - Avoided critical behavior in dynamically forced wetting. AB - A solid object can be coated by a nonwetting liquid since a receding contact line cannot exceed a critical speed. In this Letter we study the dynamical wetting transition at which a liquid film gets deposited by withdrawing a vertical plate out of a liquid reservoir. It has recently been predicted that this wetting transition is critical with diverging time scales and coincides with the disappearance of stationary menisci. We demonstrate experimentally and theoretically that the transition is due to the formation of a solitary wave, well below the critical point. As a consequence, relaxation times remain finite at threshold. The structure of the liquid deposited on the plate involves a capillary ridge that does not trivially match the Landau-Levich film. PMID- 16712303 TI - Shock-wave mach-reflection slip-stream instability: a secondary small-scale turbulent mixing phenomenon. AB - Theoretical and experimental research, on the previously unresolved instability occurring along the slip stream of a shock-wave Mach reflection, is presented. Growth rates of the large-scale Kelvin-Helmholtz shear flow instability are used to model the evolution of the slip-stream instability in ideal gas, thus indicating secondary small-scale growth of the Kelvin-Helmholtz instability as the cause for the slip-stream thickening. The model is validated through experiments measuring the instability growth rates for a range of Mach numbers and reflection wedge angles. Good agreement is found for Reynolds numbers of Re 2 x 10(4). This work demonstrates, for the first time, the use of large-scale models of the Kelvin-Helmholtz instability in modeling secondary turbulent mixing in hydrodynamic flows, a methodology which could be further implemented in many important secondary mixing processes. PMID- 16712305 TI - Polarized proton collisions at 205 GeV at RHIC. AB - The Brookhaven Relativistic Heavy Ion Collider (RHIC) has been providing collisions of polarized protons at a beam energy of 100 GeV since 2001. Equipped with two full Siberian snakes in each ring, polarization is preserved during acceleration from injection to 100 GeV. However, the intrinsic spin resonances beyond 100 GeV are about a factor of 2 stronger than those below 100 GeV making it important to examine the impact of these strong intrinsic spin resonances on polarization survival and the tolerance for vertical orbit distortions. Polarized protons were first accelerated to the record energy of 205 GeV in RHIC with a significant polarization measured at top energy in 2005. This Letter presents the results and discusses the sensitivity of the polarization survival to orbit distortions. PMID- 16712306 TI - Nontrapping arrest of Langmuir wave damping near the threshold amplitude. AB - Evolution of a Langmuir wave is studied numerically for finite amplitudes slightly above the threshold which separates damping from nondamping cases. Arrest of linear damping is found to be a second-order effect due to ballistic evolution of perturbations, resonant power transfer between field and particles, and organization of phase space into a positive slope for the average distribution function f(av) around the resonant wave phase speed nu(ph). Near the threshold trapping in the wave potential does not arrest damping or saturate the subsequent growth phase. PMID- 16712307 TI - Kinetics of the collisionless expansion of spherical nanoplasmas. AB - The collisionless expansion of spherical plasmas composed of cold ions and hot electrons is analyzed using a novel kinetic model, with special emphasis on the influence of the electron dynamics. Simple, general laws are found, relating the relevant expansion features to the initial conditions of the plasma, determined from a single dimensionless parameter. A transition is identified in the behavior of the ion energy spectrum, which is monotonic only for high electron temperatures, otherwise exhibiting a local peak far from the cutoff energy. PMID- 16712308 TI - Heating of the solar corona by dissipative Alfven solitons. AB - Solar photospheric convection drives myriads of dissipative Alfven solitons (hereinafter called alfvenons) capable of accelerating electrons and ions to energies of hundreds of keV and producing the x-ray corona. Alfvenons are exact solutions of two-fluid equations for a collisionless plasma and represent natural accelerators for conversion of the electromagnetic energy flux driven by convective flows into kinetic energy of charged particles in space and astrophysical plasmas. Their properties have been experimentally verified in the magnetosphere, where they accelerate auroral electrons to tens of keV. PMID- 16712288 TI - Search for scalar bottom quarks from gluino decays in collisions at. AB - We searched for scalar bottom quarks 156 pb(-1) of pp collisions at radicalS = 1.96 recorded by the Collider Detector at Fermilab II experiment at the Tevatron. Scalar bottom quarks can be produced from gluino decays in -parity conserving models of supersymmetry when the mass of the gluino exceeds that of the scalar bottom quark. Then, a scalar bottom quark can decay into a bottom quark and a neutralino. To search for this scenario, we investigated events with large missing transverse energy and at least three jets, two or more of which were identified as containing a secondary vertex from the hadronization of quarks. We found four candidate events, where 2.6 +/- 0.7 are expected from standard model processes, and placed 95% confidence level lower limits on gluino and scalar bottom quark masses of up to 280 and 240 GeV/c(2), respectively. PMID- 16712309 TI - Coherent structure formation in turbulent thermal superfluids. AB - By means of numerical calculations, we show that in turbulent thermal superfluids the normal fluid induces coherent bundles of quantized line vortices in the superfluid. These filamentary structures are formed in between the normal fluid vortices, acquiring eventually comparable circulation. They are self-stretched and evolve according to self-regulating dynamics. Their spectrum mimics the normal fluid spectrum with the mutual friction force exciting the large scales and damping the small scales. Strongly interacting triads of them merge sporadically into stronger, braided vortex filaments, inducing strong fluctuations in the system's energetics. A theoretical account of the system's statistical mechanics is proposed. PMID- 16712310 TI - Point defect concentrations in metastable Fe-C alloys. AB - Point defect species and concentrations in metastable Fe-C alloys are determined using density functional theory and a constrained free-energy functional. Carbon interstitials dominate unless iron vacancies are in significant excess, whereas excess carbon causes greatly enhanced vacancy concentration. Our predictions are amenable to experimental verification; they provide a baseline for rationalizing complex microstructures known in hardened and tempered steels, and by extension other technological materials created by or subjected to extreme environments. PMID- 16712311 TI - Slow crack propagation in heterogeneous materials. AB - Statistics and thermally activated dynamics of crack nucleation and propagation in a two-dimensional heterogeneous material containing quenched randomly distributed defects are studied theoretically. Using the generalized Griffith criterion we derive the equation of motion for the crack tip position accounting for dissipation, thermal noise, and the random forces arising from the defects. We find that aggregations of defects generating long-range interaction forces (e.g., clouds of dislocations) lead to anomalously slow creep of the crack tip or even to its complete arrest. We demonstrate that heterogeneous materials with frozen defects contain a large number of arrested microcracks and that their fracture toughness is enhanced to the experimentally accessible time scales. PMID- 16712312 TI - Two-step crystallization kinetics in colloidal hard-sphere systems. AB - The crystallization kinetics of colloidal hard spheres was studied using a special Bragg spectrometer with high sensitivity. In contrast with the classical scenario we observe a two-step nucleation process: the number of crystallites increases slowly at early times, followed by a dramatic reduction at intermediate times, prior to undergoing a rapid increase at late times. We explain these results in terms of a polydispersity limited growth of crystallites, where the crystallization at early times is governed by local fractionation processes, leading to a long delay prior to final crystallization. PMID- 16712314 TI - Energetic surface smoothing of complex metal-oxide thin films. AB - A novel energetic smoothing mechanism in the growth of complex metal-oxide thin films is reported from in situ kinetic studies of pulsed laser deposition of on , using x-ray reflectivity. Below 50% monolayer coverage, prompt insertion of energetic impinging species into small-diameter islands causes them to break up to form daughter islands. This smoothing mechanism therefore inhibits the formation of large-diameter 2D islands and the seeding of 3D growth. Above 50% coverage, islands begin to coalesce and their breakup is thereby suppressed. The energy of the incident flux is instead rechanneled into enhanced surface diffusion, which leads to an increase in the effective surface temperature of DeltaT approximately 500 K. These results have important implications on optimal conditions for nanoscale device fabrication using these materials. PMID- 16712313 TI - Oxygen-driven unzipping of graphitic materials. AB - Optical microscope images of graphite oxide (GO) reveal the occurrence of fault lines resulting from the oxidative processes. The fault lines and cracks of GO are also responsible for their much smaller size compared with the starting graphite materials. We propose an unzipping mechanism to explain the formation of cracks on GO and cutting of carbon nanotubes in an oxidizing acid. GO unzipping is initiated by the strain generated by the cooperative alignment of epoxy groups on a carbon lattice. We employ two small GO platelets to show that through the binding of a new epoxy group or the hopping of a nearby existing epoxy group, the unzipping process can be continued during the oxidative process of graphite. The same epoxy group binding pattern is also likely to be present in an oxidized carbon nanotube and cause its breakup. PMID- 16712315 TI - Scaling theory and morphometrics for a coarsening multiscale surface, via a principle of maximal dissipation. AB - We consider the coarsening dynamics of multiscale solutions to a dissipative singularly perturbed partial differential equation which models the evolution of a thermodynamically unstable crystalline surface. The late-time leading-order behavior of solutions is identified, through the asymptotic expansion of a maximal-dissipation principle, with a completely faceted surface governed by an intrinsic dynamical system. The properties of the resulting piecewise-affine dynamic surface predict the scaling law L(Mu) approximately t(1/3), for the growth in time of a characteristic morphological length scale L(Mu). A novel computational geometry tool which directly simulates a million-facet piecewise affine dynamic surface is also introduced. Our computed data are consistent with the dynamic scaling hypothesis, and we report a variety of associated morphometric scaling functions. PMID- 16712316 TI - Quantum degeneracy of microcavity polaritons. AB - We investigate experimentally one of the main features of a quantum fluid constituted by exciton polaritons in a semiconductor microcavity, that is, quantum degeneracy of a macroscopic fraction of the particles. We show that resonant pumping allows us to create a macroscopic population of polaritons in one quantum state. Furthermore, we demonstrate that parametric polariton scattering results in the transfer of a macroscopic population of polariton from one single quantum state into another one. Finally, we briefly outline a simple method which provides direct evidence of the first-order spatial coherence of the transferred population. PMID- 16712317 TI - Drastic change in transport of entropy with quadrupolar ordering in. AB - The antiferroquadrupolar ordering of is explored by probing thermal and thermoelectric transport. The lattice thermal conductivity drastically increases with the ordering, as a consequence of a large drop in carrier concentration and a strong electron-phonon coupling. The low level of carrier density in the ordered state is confirmed by the anomalously large values of the Seebeck and Nernst coefficients. The results are reminiscent of and suggest that both belong to the same class of partial metal-insulator transitions. The magnitude of the Nernst coefficient, larger than in any other metal, indicates a new route for Ettingshausen cooling at Kelvin temperatures. PMID- 16712318 TI - Using a quantum dot as a high-frequency shot noise detector. AB - We present the experimental realization of a quantum dot (QD) operating as a high frequency noise detector. Current fluctuations produced in a nearby quantum point contact (QPC) ionize the QD and induce transport through excited states. The resulting transient current through the QD represents our detector signal. We investigate its dependence on the QPC transmission and voltage bias. We observe and explain a quantum threshold feature and a saturation in the detector signal. This experimental and theoretical study is relevant in understanding the backaction of a QPC used as a charge detector. PMID- 16712319 TI - Orientation-dependent transparency of metallic interfaces. AB - As devices are reduced in size, interfaces start to dominate electrical transport, making it essential to be able to describe reliably how they transmit and reflect electrons. For a number of nearly perfectly lattice-matched materials, we calculate from first principles the dependence of the interface transparency on the crystal orientation. Quite remarkably, the largest anisotropy is predicted for interfaces between the prototype free-electron materials silver and aluminum, for which a massive factor of 2 difference between (111) and (001) interfaces is found. PMID- 16712321 TI - Vibration-induced Kondo tunneling through metal-organic complexes with even electron occupation number. AB - We investigate transport through a mononuclear transition-metal complex with strong tunnel coupling to two electrodes. The ground state of this molecule is a singlet, while the first excited state is a triplet. We show that a modulation of the tunnel-barrier due to a molecular distortion which couples to the tunneling induces a Kondo-effect, provided the discrete vibrational energy compensates the singlet-triplet gap. We discuss the single-phonon and two-phonon-assisted cotunneling and possible experimental realization of the theory. PMID- 16712320 TI - Electrical detection of spin accumulation at a ferromagnet-semiconductor interface. AB - We show that the accumulation of spin-polarized electrons at a forward-biased Schottky tunnel barrier between Fe and -GaAs can be detected electrically. The spin accumulation leads to an additional voltage drop across the barrier that is suppressed by a small transverse magnetic field, which depolarizes the spins in the semiconductor. The dependence of the electrical accumulation signal on magnetic field, bias current, and temperature is in good agreement with the predictions of a drift-diffusion model for spin-polarized transport. PMID- 16712322 TI - Spectroscopy of the Kondo problem in a box. AB - Motivated by experiments on double quantum dots, we study the problem of a single magnetic impurity confined in a finite metallic host. We prove an exact theorem for the ground state spin, and use analytic and numerical arguments to map out the spin structure of the excitation spectrum of the many-body Kondo-correlated state, throughout the weak to strong coupling crossover. These excitations can be probed in a simple tunneling-spectroscopy transport experiment; for that situation we solve rate equations for the conductance. PMID- 16712323 TI - Spin-filtered edge states and quantum Hall effect in graphene. AB - Electron edge states in graphene in the quantum Hall effect regime can carry both charge and spin. We show that spin splitting of the zeroth Landau level gives rise to counterpropagating modes with opposite spin polarization. These chiral spin modes lead to a rich variety of spin current states, depending on the spin flip rate. A method to control the latter locally is proposed. We estimate Zeeman spin splitting enhanced by exchange, and obtain a spin gap of a few hundred Kelvin. PMID- 16712324 TI - Electron transport in a double quantum dot governed by a nuclear magnetic field. AB - We investigate theoretically electron transfer in a double dot in a situation where spin blockade is lifted by nuclear magnetic field: this has been recently achieved in experiment [F. Koppens, Science 309, 1346 (2005)]. We show that for a given realization of nuclear magnetic field spin blockade can be restored by tuning external magnetic field; this may be useful for quantum manipulation of the device. PMID- 16712325 TI - Surface melting of the vortex lattice. AB - We discuss the effect of an (ab) surface on the melting transition of the pancake vortex lattice in a layered superconductor within a density functional theory approach. Both discontinuous and continuous surface melting are predicted for this system, although the latter scenario occupies the major part of the low field phase diagram. The formation of a quasiliquid layer below the bulk melting temperature inhibits the appearance of a superheated solid phase, yielding an asymmetric hysteretic behavior which has been seen in experiments. PMID- 16712326 TI - Effect of irradiation-induced disorder on the conductivity and critical temperature of the organic superconductor kappa-(BEDT-TTF)2Cu(SCN)2. AB - We have introduced defects into clean samples of the organic superconductor kappa (BEDT-TTF)(2)Cu(SCN)(2) in order to determine their effect on the temperature dependence of the interlayer conductivity and the critical temperature T(c). We find a violation of Matthiessen's rule that can be explained by a model of involving a defect-assisted interlayer channel which acts in parallel with the bandlike conductivity. We observe an unusual dependence of T(c) on residual resistivity, inconsistent with the generalized Abrikosov-Gor'kov theory for an order parameter with a single component, providing an important constraint on models of the superconductivity in this material. PMID- 16712327 TI - Enhanced macroscopic quantum tunneling in Bi2Sr2CaCu2O8 + delta intrinsic Josephson-junction stacks. AB - We have investigated macroscopic quantum tunneling in Bi(2)Sr(2)CaCu(2)O(8 + delta) intrinsic Josephson junctions at millikelvin temperatures using microwave irradiation. Measurements show that the escape rate for uniformly switching stacks of Nu junctions is about Nu(2) times higher than that of a single junction having the same plasma frequency. We argue that this gigantic enhancement of the macroscopic quantum tunneling rate in stacks is boosted by current fluctuations which occur in the series array of junctions loaded by the impedance of the environment. PMID- 16712328 TI - Field-induced order and spin waves in the pyrochlore antiferromagnet Tb2Ti207. AB - High resolution time-of-flight neutron scattering measurements on Tb(2)Ti(2)0(7) reveal a rich low temperature phase diagram in the presence of a magnetic field applied along [110]. In zero field at T = 0.4 K, Tb(2)Ti(2)0(7) is a highly correlated cooperative paramagnet with disordered spins residing on a pyrochlore lattice of corner-sharing tetrahedra. Application of a small field condenses much of the magnetic diffuse scattering, characteristic of the disordered spins, into a new Bragg peak characteristic of a polarized paramagnet. At higher fields, a magnetically ordered phase is induced, which supports spin wave excitations indicative of continuous, rather than Ising-like, spin degrees of freedom. PMID- 16712329 TI - Interferometric detection of spin-polarized transport in the depletion layer of a metal-GaAs Schottky barrier. AB - It is shown that the Kerr rotation of spin-polarized electrons is modulated by the distance of the electrons from the sample surface. Time-resolved Kerr rotation of optically excited spin-polarized electrons in the depletion layer of n-doped GaAs displays fast oscillations that originate from interference between the light reflected from the semiconductor surface and from the front of the electron distribution moving into the semiconductor. Using this effect, the dynamics of the photogenerated charge carriers in the depletion layer of the biased Schottky barrier is measured. PMID- 16712330 TI - Local spin polarization at surfaces probed by hollow atoms. AB - The relaxation of hollow atoms produced by slow multiply charged ions impinging on surfaces produces characteristic Auger electron spectra. These spectra, which serve as fingerprints of the interaction, can be used to probe local spin ordering at surfaces by relating changes in the intensities of different spin states to local spin polarization at the surface. The area from which the electrons are captured is of the order of a few Angstrom(2), only. The potential of the method is illustrated by He(2+) and N(6+) ions interacting with a ferromagnetic Ni(110) crystal. From the Auger spectra we determine a spin polarization of approximately 90% at room temperature. PMID- 16712331 TI - Structure and dynamics of liquid crystalline pattern formation in drying droplets of DNA. AB - We investigate the formation of ringlike deposits in drying drops of DNA. In analogy with the colloidal "coffee rings," DNA is transported to the perimeter by the capillary flow. At the droplet edge, however, DNA forms a lyotropic liquid crystal (LC) with concentric chain orientations to minimize the LC elastic energy. During the final stages of drying, the contact line retracts, and the radial stress causes undulations at the rim that propagate inward through the LC and form a periodic zigzag structure. We examine the phenomenon in terms of a simple model based on LC elasticity. PMID- 16712332 TI - Short range attraction between two similarly charged silica surfaces. AB - Using an atomic force microscope we measure the interaction between two identically charged silica surfaces in the presence of a saline solution. For pure NaCl the interaction is always repulsive. Upon addition of cobalt hexamine ions, Co(NH(3))(6)(+3), the repulsion is gradually suppressed and a pronounced attraction develops at distances much shorter than the screening length. Higher concentrations of cobalt hexamine turn the attraction back into repulsion. Measurements of surface charge renormalization by the trivalent cations provide their surface density and their association constant to the negatively charged silica surface. These estimates tend to exclude interaction between two condensed Wigner crystals as an explanation for the attraction. PMID- 16712333 TI - Viscous water meniscus under nanoconfinement. AB - A dramatic transition in the mechanical properties of water is observed at the nanometer scale. For a water meniscus formed between two hydrophilic surfaces in the attractive region, with < or = 1 nm interfacial separation, the measured viscosity is 7 orders of magnitude greater than that of bulk water at room temperature. Grand canonical Monte Carlo simulations reveal enhancement in the tetrahedral structure and in the number of hydrogen bonds to the surfaces as a source for the high viscosity; this results from a cooperative effect of hydrogen bonding of water molecules to both hydrophilic surfaces. PMID- 16712334 TI - Thermodynamics predicts how confinement modifies the dynamics of the equilibrium hard-sphere fluid. AB - We study how confining the equilibrium hard-sphere fluid to restrictive one- and two-dimensional channels with smooth interacting walls modifies its structure, dynamics, and entropy using molecular dynamics and transition-matrix Monte Carlo simulations. Although confinement strongly affects local structuring, the relationships between self-diffusivity, excess entropy, and average fluid density are, to an excellent approximation, independent of channel width or particle-wall interactions. Thus, thermodynamics can be used to predict how confinement impacts dynamics. PMID- 16712335 TI - Universality in structure and elasticity of polymer-nanoparticle gels. AB - We propose a combination of polymer field theory and off-lattice computer simulations to study polymer-bridged gelation in polymer-nanoparticle mixtures. We use this method to study the structure of gels formed in attractive polymer colloid systems. Our results indicate that such gels exhibit a universal structure with a fractal dimension d(f) approximately 2.5 characteristic of random percolation. By mapping to an affine-network model, the enhancement in elastic moduli is predicted to follow a critical exponent nu(eta) approximately 1.8 characteristic of the resistor network percolation. We analyze selected experimental results to suggest the existence of a universality class corresponding to our results. PMID- 16712336 TI - Fluctuation-induced casimir forces in granular fluids. AB - We numerically investigate the behavior of driven noncohesive granular media and find that two fixed large intruder particles, immersed in a sea of small particles, experience, in addition to a short-range depletion force, a long-range repulsive force. The observed long-range interaction is fluctuation-induced and we propose a mechanism similar to the Casimir effect that generates it: The hydrodynamic fluctuations are geometrically confined between the intruders, producing an unbalanced renormalized pressure. An estimation based on computing the possible Fourier modes explains the repulsive force and is in qualitative agreement with the simulations. PMID- 16712337 TI - Triboelectrification and razorbacks: geophysical patterns produced in dry grains. AB - Electrostatic interactions between particles can dramatically affect granular flows, creating industrial safety and handling problems [K. N. Palmer, (Chapman and Hall, London, 1973), pp. 388-389]. We present experimental data demonstrating that charging of grains can also cause spontaneous self-assembly that may generate lasting geological patterns under arid conditions. Paradoxically, we find that grains that tribocharge enough to produce small explosions, ejecting grains meters into the air, leave little net charge on grains. Rather, grains charge into strongly heterogeneous polar clusters. These assemble into stereotyped residual structures that resemble geological features, for example, razorbacks observed on Mars ["The Razorback Mystery," July 16, 2004, http://www.jpl.nasa.gov/missions/mer/images.cfm?id=701]. PMID- 16712338 TI - Maximum-entropy closures for kinetic theories of neuronal network dynamics. AB - We analyze (1 + 1)D kinetic equations for neuronal network dynamics, which are derived via an intuitive closure from a Boltzmann-like equation governing the evolution of a one-particle (i.e., one-neuron) probability density function. We demonstrate that this intuitive closure is a generalization of moment closures based on the maximum-entropy principle. By invoking maximum-entropy closures, we show how to systematically extend this kinetic theory to obtain higher-order, kinetic equations and to include coupled networks of both excitatory and inhibitory neurons. PMID- 16712339 TI - Wringing out DNA. AB - The chiral nature of DNA plays a crucial role in cellular processes. Here we use magnetic tweezers to explore one of the signatures of this chirality, the coupling between stretch and twist deformations. We show that the extension of a stretched DNA molecule increases linearly by 0.42 nm per excess turn applied to the double helix. This result contradicts the intuition that DNA should lengthen as it is unwound and get shorter with overwinding. We then present numerical results of energy minimizations of torsionally restrained DNA that display a behavior similar to the experimental data and shed light on the molecular details of this surprising effect. PMID- 16712340 TI - Dynamics of surfactant-driven fracture of particle rafts. AB - We investigate the dynamic fracture of a close-packed monolayer of particles, or particle raft, floating at a liquid-gas interface induced by the localized addition of surfactant. Unusually for a two-dimensional solid, our experiments show that the speed of crack propagation here is not affected by the elastic properties of the raft. Instead it is controlled by the rate at which surfactant is advected to the crack tip by means of the induced Marangoni flows. Further, the velocity of propagation is not constant in time and the length of the crack scales as t(3/4). More broadly, this surfactant-induced rupture of interfacial rafts suggests ways to manipulate them for applications. PMID- 16712341 TI - Thermal undulations of lipid bilayers relax by intermonolayer friction at submicrometer length scales. AB - The time correlation functions of the thermal undulations of a lipid membrane have been studied by molecular dynamics simulations of a coarse-grained bilayer model. We observe a double-exponential decay, with relaxation rates in good agreement with the theory by Seifert and Langer, [Europhys. Lett. 23, 71 (1993)]. Intermonolayer friction resulting from local velocity differences between the two monolayers is shown to be the dominant dissipative mechanism for fluctuations with wave lengths below approximately -0.1 microm. PMID- 16712342 TI - Thermal effects in the splashing of drops under a reduced pressure Environment. PMID- 16712344 TI - Positronium collapse and the maximum magnetic field in pure QED. AB - A maximum value for the magnetic field is determined, which provides the full compensation of the positronium rest mass by the binding energy in the maximum symmetry state and disappearance of the energy gap separating the electron positron system from the vacuum. The compensation becomes possible owing to the falling to the center phenomenon. The maximum magnetic field may be related to the vacuum and describe its structure. PMID- 16712345 TI - Bose-fermi mixtures in a three-dimensional optical lattice. AB - We have studied mixtures of fermionic (40)K and bosonic (87)Rb quantum gases in a three-dimensional optical lattice. We observe that an increasing admixture of the fermionic species diminishes the phase coherence of the bosonic atoms as measured by studying both the visibility of the matter wave interference pattern and the coherence length of the bosons. Moreover, we find that the attractive interactions between bosons and fermions lead to an increase of the boson density in the lattice which we measure by studying three-body recombination in the lattice. In our data, we do not observe three-body loss of the fermionic atoms. An analysis of the thermodynamics of a noninteracting Bose-Fermi mixture in the lattice suggests a mechanism for sympathetic cooling of the fermions in the lattice. PMID- 16712346 TI - Localization of bosonic atoms by fermionic impurities in a three-dimensional optical lattice. AB - We observe a localized phase of ultracold bosonic quantum gases in a 3 dimensional optical lattice induced by a small contribution of fermionic atoms acting as impurities in a Fermi-Bose quantum gas mixture. In particular, we study the dependence of this transition on the fermionic (40)K impurity concentration by a comparison to the corresponding superfluid to Mott-insulator transition in a pure bosonic (87)Rb gas and find a significant shift in the transition parameter. The observed shift is larger than expected based on a simple mean-field argument, which indicates that disorder-related effects play a significant role. PMID- 16712347 TI - Integrative models for asymmetric fermi superfluids: emergence of a new exotic pairing phase. AB - We introduce an exactly solvable model to study the competition between the Larkin-Ovchinnikov-Fulde-Ferrell (LOFF) and breached-pair superfluid in strongly interacting ultracold asymmetric Fermi gases. One can thus investigate homogeneous and inhomogeneous states on equal footing and establish the quantum phase diagram. For certain values of the filling and the interaction strength, the model exhibits a new stable exotic pairing phase which combines an inhomogeneous state with an interior gap to pair excitations. It is proven that this phase is the exact ground state in the strong-coupling limit, while numerical examples in finite lattices show that also at finite interaction strength it can have lower energy than the breached-pair or LOFF states. PMID- 16712348 TI - Observation of strong quantum depletion in a gaseous Bose-Einstein condensate. AB - We studied quantum depletion in a gaseous Bose-Einstein condensate. An optical lattice enhanced the atomic interactions and modified the dispersion relation resulting in strong quantum depletion. The depleted fraction was directly observed as a diffuse background in the time-of-flight images. Bogoliubov theory provides a semiquantitative description for our observations of depleted fractions in excess of 50%. PMID- 16712349 TI - Vortex-Peierls states in optical lattices. AB - We show that vortices, induced in cold atom superfluids in optical lattices, may order in a novel vortex-Peierls ground state. In such a state vortices do not form a simple lattice but arrange themselves in clusters, within which the vortices are partially delocalized, tunneling between classically degenerate configurations. We demonstrate that this exotic quantum many-body state is selected by an order-from-disorder mechanism for a special combination of the vortex filling and lattice geometry that has a macroscopic number of classically degenerate ground states. PMID- 16712350 TI - High-field fractional quantum Hall effect in optical lattices. AB - We consider interacting bosonic atoms in an optical lattice subject to a large simulated magnetic field. We develop a model similar to a bilayer fractional quantum Hall system valid near simple rational numbers of magnetic flux quanta per lattice cell. Then we calculate its ground state, magnetic lengths, fractional fillings, and find unexpected sign changes in the Hall current. Finally we study methods for detecting these novel features via shot noise and Hall current measurements. PMID- 16712351 TI - Deutsch-jozsa algorithm using triggered single photons from a single quantum dot. AB - We demonstrate a two-qubit Deutsch-Jozsa algorithm with single photons from a single InP quantum dot. The qubits are implemented via the spatial mode and the polarization of a single photon. Our photon source is operated both under continuous and pulsed excitation, the latter allowing deterministic quantum logic by generating photons on demand with a strong suppression of two-photon events. The computation reached a success probability of up to 79%. We also exploit the concept of decoherence-free subspaces that helps to make our experimental setup robust against sources of phase noise. PMID- 16712352 TI - Brownian flights over a fractal nest and first-passage statistics on irregular surfaces. AB - The diffusive motion of Brownian particles near irregular interfaces plays a crucial role in various transport phenomena in nature and industry. Most diffusion-reaction processes in confining interfacial systems involve a sequence of Brownian flights in the bulk, connecting successive hits with the interface (Brownian bridges). The statistics of times and displacements separating two interface encounters are then determinant in the overall transport. We present a theoretical and numerical analysis of this complex first-passage problem. We show that the bridge statistics is directly related to the Minkowski content of the surface within the usual diffusion length. In the case of self-similar or self affine interfaces, we show and check numerically that the bridge statistics follows power laws with exponents depending directly on the surface fractal dimension. PMID- 16712353 TI - Simple model for active nematics: quasi-long-range order and giant fluctuations. AB - We propose a simple microscopic model for active nematic particles similar in spirit to the Vicsek model for self-propelled polar particles. In two dimensions, we show that this model exhibits a Kosterlitz-Thouless-like transition to quasi long-range orientational order and that in this nonequilibrium context, the ordered phase is characterized by giant density fluctuations, in agreement with the predictions of Ramaswamy et al. PMID- 16712354 TI - Loop updates for quantum Monte Carlo simulations in the canonical ensemble. AB - We present a new nonlocal updating scheme for quantum Monte Carlo simulations, which conserves particle number and other symmetries. It allows exact symmetry projection and direct evaluation of the equal-time Green's function and other observables in the canonical ensemble. The method is applicable to a wide variety of systems. We show results for bosonic atoms in optical lattices, neutron pairs in atomic nuclei, and electron pairs in ultrasmall superconducting grains. PMID- 16712355 TI - Zurek-Kibble mechanism for the spontaneous vortex formation in Nb-Al/Al(ox)/Nb Josephson tunnel junctions: new theory and experiment. AB - New scaling behavior has been both predicted and observed in the spontaneous production of fluxons in quenched Nb-Al/Al(ox)/Nb annular Josephson tunnel junctions (JTJs) as a function of the quench time, tau(Q). The probability f(1) to trap a single defect during the normal-metal-superconductor phase transition clearly follows an allometric dependence on tau(Q) with a scaling exponent sigma = 0.5, as predicted from the Zurek-Kibble mechanism for realistic JTJs formed by strongly coupled superconductors. This definitive experiment replaces one reported by us earlier, in which an idealized model was used that predicted sigma = 0.25, commensurate with the then much poorer data. Our experiment remains the only condensed matter experiment to date to have measured a scaling exponent with any reliability. PMID- 16712356 TI - Rayleigh-Plateau and Gregory-Laflamme instabilities of black strings. AB - Many and very general arguments indicate that the event horizon behaves as a stretched membrane. We explore this analogy by associating the Gregory-Laflamme instability of black strings with a classical membrane instability known as the Rayleigh-Plateau instability. We show that the key features of the black string instability can be reproduced using this viewpoint. In particular, we get good agreement for the threshold mode in all dimensions and exact agreement for large spacetime dimensionality. The instability time scale is also well described within this model, as well as the dimensionality dependence. It also predicts that general nonaxisymmetric perturbations are stable. We further argue that the instability of ultraspinning black holes follows from this model. PMID- 16712357 TI - Holographic derivation of entanglement entropy from the anti-de Sitter space/conformal field theory correspondence. AB - A holographic derivation of the entanglement entropy in quantum (conformal) field theories is proposed from anti-de Sitter/conformal field theory (AdS/CFT) correspondence. We argue that the entanglement entropy in d + 1 dimensional conformal field theories can be obtained from the area of d dimensional minimal surfaces in AdS(d+2), analogous to the Bekenstein-Hawking formula for black hole entropy. We show that our proposal agrees perfectly with the entanglement entropy in 2D CFT when applied to AdS(3). We also compare the entropy computed in AdS(5)XS(5) with that of the free N=4 super Yang-Mills theory. PMID- 16712359 TI - Compact multigluonic scattering amplitudes with heavy scalars and fermions. AB - Combining the Berends-Giele and on-shell recursion relations we obtain an extremely compact expression for the scattering amplitude of a complex massive scalar-antiscalar pair and an arbitrary number of positive helicity gluons. This is one of the basic building blocks for constructing other helicity configurations from recursion relations. We also show explicitly that the scattering amplitude of massive fermions to gluons, all with positive helicity, is proportional to the scalar one, confirming in this way the recently advocated SUSY-like Ward identities relating both amplitudes. PMID- 16712362 TI - Multicolored coherent population trapping in a system using phase modulated fields. AB - We demonstrate the simultaneous occurrence of coherent population trapping at a series of frequencies separated by modulation frequency of phase-modulated fields. The two arms of the system are coupled to two phase-modulated fields and the response of the atomic system to such fields is calculated nonperturbatively. A judicious choice of modulation characteristics allows one to selectively switch on or off the occurrence of coherent population trapping at specific frequencies. A new technique is developed to compute two-dimensional tridiagonal matrix equations. This generalized technique provides the vital methodology needed to calculate the response of such systems in the strong modulation regime and for arbitrary field strengths. PMID- 16712361 TI - Vanishing N = 20 shell gap: study of excited states in (27,28)Ne. AB - This Letter reports on the (1)H((28)Ne, (28)Ne) and (1)H((28)Ne, (27)Ne) reactions studied at intermediate energy using a liquid hydrogen target. From the cross section populating the first 2(+) excited state of (28)Ne, and using the previously determined BE(2) value, the neutron quadrupole transition matrix element has been calculated to be M(n)=13.8 +/- 3.7 fm(2). In the neutron knockout reaction, two low-lying excited states were populated in (27)Ne. Only one of them can be interpreted by the sd shell model while the additional state may intrude from the fp shell. These experimental observations are consistent with the presence of fp shell configurations at low excitation energy in (27,28)Ne nuclei caused by a vanishing N=20 shell gap at Z=10. PMID- 16712363 TI - Phase and amplitude retrieval in ghost diffraction from field-correlation measurements. AB - We report the results of experiments about the inversion of ghost diffraction with pseudothermal light. A complete retrieval of the complex transmission function of planar transparencies, illuminated by spatially incoherent, quasimonochromatic light, is achieved. This is obtained by measuring the field (instead of the intensity) correlation function. In particular, the determination of the phase of the correlation function is made particularly easy and robust by the use of a suitably modified Young interferometer. The presented results refer to the cases of a clear slit and a phase step. PMID- 16712364 TI - Extended quasimodes within nominally localized random waveguides. AB - We have measured the spatial and spectral dependence of the microwave field inside an open absorbing waveguide filled with randomly juxtaposed dielectric slabs in the spectral region in which the average level spacing exceeds the typical level width. Whenever lines overlap in the spectrum, the field exhibits multiple peaks within the sample. Only then is substantial energy found beyond the first half of the sample. When the spectrum throughout the sample is decomposed into a sum of Lorentzian lines plus a broad background, their central frequencies and widths are found to be essentially independent of position. Thus, this decomposition provides the electromagnetic quasimodes underlying the extended field in nominally localized samples. These quasimodes may exhibit multiple peaks in space when they overlap spectrally. PMID- 16712358 TI - Improved search for nu(mu) --> nu(e) oscillation in a long-baseline accelerator experiment. AB - We performed an improved search for nu(mu) --> nu(e) oscillation with the KEK to Kamioka (K2K) long-baseline neutrino oscillation experiment, using the full data sample of 9.2 x 10(19) protons on target. No evidence for a nu(e) appearance signal was found, and we set bounds on the nu(mu) --> nu(e) oscillation parameters. At Deltam(2)=2.8 x 10(-3) eV(2), the best-fit value of the K2Knu(mu) disappearance analysis, we set an upper limit of sin(2)2theta(mue) < 0.13 at a 90% confidence level. PMID- 16712360 TI - Observation of Psi(3770)-->gammachi(c1)-->gammagammaJ/Psi. AB - From e(+)e(-) collision data acquired with the CLEO detector at the Cornell Electron Storage Ring, we observe the non-DD(_) decay Psi(3770))-->gammachi(c1) with a statistical significance of 6.6 standard deviations, using the two-photon cascades to J/Psi and J/Psi-->l(+)l(-). We determine sigma(e(=)e(-)- >Psi(3770))xBeta(Psi(3770)-->gammachi(c1))=(18.0 +/- 3.3 +/- 2.5) pb and branching fraction Beta(Psi(3770)-->gammachi(c1)=(2.8 +/- 0.5+/-0.4) x 10(-3). We set 90% C.L. upper limits for the transition to chi(c2) (chi(c0)): sigma x Beta<5.7 pb (<282 pb) and Beta<0.9 x 10(-3) (<44 x 10(-3)). We also determine Gamma(Psi(3770)gammachi(c1))/Gamma(Psi(3770)-->pi(+)pi(-)J/Psi)=1.5 +/- 0.3 +/- 0.3 (>1.0 at 90% C.L.), which bears upon the interpretation of X(3872). PMID- 16712365 TI - Novel heat conduction model for bridging different space and time scales. AB - This exposition describes a novel heat transport model and an underlying unified theory emanating from the physics of the Boltzmann transport equation which acknowledges simultaneously the coexistence of that termed as slow processes (at low energies) and fast processes (at high energies) as heat carriers while describing the evolution of heat transport characteristics spanning both spatial scales (characterizing ballistic to diffusive limits), and also time scales (characterizing finite to infinite heat propagation speeds). PMID- 16712366 TI - Large-scale anisotropy in scalar turbulence. AB - The effect of anisotropy on the statistics of a passive tracer transported by a turbulent flow is investigated. We show that under broad conditions an arbitrarily small amount of anisotropy propagates to the large scales where it eventually dominates the structure of the concentration field. This result is obtained analytically in the framework of an exactly solvable model and confirmed by numerical simulations of scalar transport in two-dimensional turbulence. PMID- 16712367 TI - Evolution of filament structures during edge-localized modes in the MAST Tokamak. AB - Edge-localized modes (ELMs) are repetitive instabilities that occur in the outer region of tokamak plasmas. This Letter provides new information on and the implications of the evolution of the filament structures observed during ELMs in the MAST tokamak. The filaments exist for the time over which particles are being released into the scrape off layer. They start off at the plasma edge rotating at the velocity of the pedestal, and then decelerate toroidally and accelerate radially outwards. As the filaments propagate radially they remain aligned with the local magnetic field line. PMID- 16712368 TI - Radiation transfer effects on the spectra of laser-generated plasmas. AB - Experimental x-ray spectra of the H-like 2p --> 1s (Lyman-alpha) doublet have been obtained using time-integrated high-resolution spectroscopy of a constrained flow, laser-generated aluminum plasma. These spectra show monotonic alteration of the relative intensities of the doublet components with distance from the target surface. Excellent agreement between experiment and theory is found only if the modeling includes both ion collisional rates between the fine-structure components of the level and, more importantly, the radiative pumping of one Lyman alpha component by the other component along the direction of the major velocity gradient (i.e., perpendicular to the direction of spectra observation). Understanding radiation transfer in plasmas with high velocity gradients is important in modeling many astrophysical objects, and this experiment acts as a benchmark for such complex calculations. PMID- 16712369 TI - Heating by an electron Bernstein wave in a spherical tokamak plasma via mode conversion. AB - The first successful high power heating of a high dielectric constant spherical tokamak plasma by an electron Bernstein wave (EBW) is reported. An EBW was excited by mode conversion (MC) of an mode cyclotron wave injected from the low magnetic field side of the TST-2 spherical tokamak. Evidence of electron heating was observed as increases in the stored energy and soft x-ray emission. The increased emission was concentrated in the plasma core region. A heating efficiency of over 50% was achieved, when the density gradient in the MC region was sufficiently steep. PMID- 16712370 TI - Interfacial segregation and electro-diffusion of dopants in superlattices. AB - A first-principles theory of interfacial segregation of dopants and defects in heterostructures is developed and applied to GAN/A1N superlattices. The results indicate that the equilibrium concentrations of a dopant at two sides of an interface may differ by up to a few orders of magnitude, depending on its chemical identity and charge state, and that these cannot be obtained from calculations for bulk constituents alone. In addition, the presence of an internal electric field in polar heterostructures induces electro-migration and accumulation of hydrogen at the appropriate interfaces. PMID- 16712371 TI - Microscopic structure and elasticity of weakly aggregated colloidal gels. AB - We directly probe the microscopic structure, connectivity, and elasticity of colloidal gels using confocal microscopy. We show that the gel is a random network of one-dimensional chains of particles. By measuring thermal fluctuations, we determine the effective spring constant between pairs of particles as a function of separation; this is in agreement with the theory for fractal chains. Long-range attractions between particles lead to freely rotating bonds, and the gel is stabilized by multiple connections among the chains. By contrast, short-range attractions lead to bonds that resist bending, with dramatically suppressed formation of loops of particles. PMID- 16712373 TI - Predicting the long-time dynamic heterogeneity in a supercooled liquid on the basis of short-time heterogeneities. AB - We report that the local Debye-Waller factor in a simulated 2D glass-forming mixture exhibits significant spatial heterogeneities and that these short-time fluctuations provide an excellent predictor of the spatial distribution of the long-time dynamic propensities. In contrast, the potential energy per particle of the inherent structure does not correlate well with the spatially distributed dynamics. PMID- 16712372 TI - Insights into the fracture mechanisms and strength of amorphous and nanocomposite carbon. AB - Tight-binding molecular dynamics simulations shed light into the fracture mechanisms and the ideal strength of tetrahedral amorphous carbon and of nanocomposite carbon containing diamond crystallites, two of the hardest materials. It is found that fracture in the nanocomposites, under tensile or shear load, occurs intergrain and so their ideal strength is similar to the pure amorphous phase. The onset of fracture takes place at weakly bonded sites in the amorphous matrix. On the other hand, the nanodiamond inclusions significantly enhance the elastic moduli, which approach those of diamond. PMID- 16712374 TI - Thermal conductance of hydrophilic and hydrophobic interfaces. AB - Using time-domain thermoreflectance, we have measured the transport of thermally excited vibrational energy across planar interfaces between water and solids that have been chemically functionalized with a self-assembled monolayer (SAM). The Kapitza length--i.e., the thermal conductivity of water divided by the thermal conductance per unit area of the interface--is analogous to the "slip length" for water flowing tangentially past a solid surface. We find that the Kapitza length at hydrophobic interfaces (10-12 nm) is a factor of 2-3 larger than the Kapitza length at hydrophilic interfaces (3-6 nm). If a vapor layer is present at the hydrophobic interface, and this vapor layer has a thermal conductivity that is comparable to bulk water vapor, then our experimental results constrain the thickness of the vapor layer to be less than 0.25 nm. PMID- 16712375 TI - Giant amplification of interfacially driven transport by hydrodynamic slip: diffusio-osmosis and beyond. AB - We demonstrate that "moderate" departures from the no-slip hydrodynamic boundary condition (hydrodynamic slip lengths in the nanometer range) can result in a very large enhancement--up to 2 orders of magnitude--of most interfacially driven transport phenomena. We study analytically and numerically the case of neutral solute diffusio-osmosis in a slab geometry to account for nontrivial couplings between interfacial structure and hydrodynamic slip. Possible outcomes are fast transport of particles in externally applied or self-generated gradient, and flow enhancement in nano- or microfluidic geometries. PMID- 16712376 TI - Stress and morphology evolution during island growth. AB - We performed a series of hybrid molecular-dynamics simulations of island growth on a substrate and monitored island stress evolution for several different island/substrate interfacial energies. Smaller (larger) interfacial energy yields islands with a stronger (weaker) compressive stress-thickness product. We present analytical results that suggest that the stress-thickness product is a linear function of the substrate coverage, with slope equal to minus the substrate surface stress, if the island is in mechanical equilibrium, and verify these results with simulation data. PMID- 16712377 TI - C(60) molecular bearings and the phenomenon of nanomapping. AB - Inspired by suggestions of C(60) "nanobearings," we have measured sliding friction on fixed and rotating C(60) layers to explore whether a lubricating effect is present. We refer to this general phenomenon as "nanomapping," whereby macroscopic attributes are mapped in a one on one fashion to nanoscale entities. Our measurements are the first to directly link friction to a documented molecular rotation state. Friction is, however, observed to be higher for rotating layers, in defiance of the ball-bearing analogy. Thus, no direct mapping of macro- to nanoscale attributes can be established. PMID- 16712378 TI - Dynamical response of nanomechanical oscillators in immiscible viscous fluid for in vitro biomolecular recognition. AB - Dynamical response of nanomechanical cantilever structures immersed in a viscous fluid is important to in vitro single-molecule force spectroscopy, biomolecular recognition of disease-specific proteins, and the study of microscopic protein dynamics. Here we study the stochastic response of biofunctionalized nanomechanical cantilever beams in a viscous fluid. Using the fluctuation dissipation theorem we derive an exact expression for the spectral density of displacement and a linear approximation for resonance frequency shift. We find that in a viscous solution the frequency shift of the nanoscale cantilever is determined by surface stress generated by biomolecular interaction with negligible contributions from mass loading due to the biomolecules. PMID- 16712379 TI - Bosonization in higher dimensions via noncommutative field theory. AB - We propose the bosonization of a many-body fermion theory in D spatial dimensions through a noncommutative field theory on a (2D-1)-dimensional space. This theory leads to a chiral current algebra over the noncommutative space and reproduces the correct perturbative Hilbert space and excitation energies for the fermions. The validity of the method is demonstrated by bosonizing a two-dimensional gas of fermions in a harmonic trap. PMID- 16712380 TI - Separation of orbital contributions to the optical conductivity of BaVS(3). AB - The correlation-driven metal-insulator transition (MIT) of BaVS(3) was studied by polarized infrared spectroscopy. In the metallic state two types of electrons coexist at the Fermi energy: the quasi-1D metallic transport of A(1g) electrons is superimposed on the isotropic hopping conduction of localized E(g) electrons. The "bad-metal" character and the weak anisotropy are the consequences of the large effective mass m(eff) approximately 7 m(e) and scattering rate Gamma > or = 160 meV of the quasiparticles in the A(1g) band. There is a pseudogap above T(MI) = 69 K, and in the insulating phase the gap follows the BCS-like temperature dependence of the structural order parameter with Delta(ch) approximately 42 meV in the ground state. The MIT is described in terms of a weakly coupled two-band model. PMID- 16712381 TI - Anomalous electronic correlations in the ground state momentum density of Al(97)Li(3). AB - We report high resolution Compton scattering measurements on an Al(97)Li(3) disordered alloy single crystal for momentum transfer along the [100], [110], and [111] symmetry directions. The results are interpreted via corresponding Korringa Kohn-Rostoker coherent potential approximation first-principles computations. By comparing spectra for Al(97)Li(3) and Al, we show that the momentum density in the alloy differs significantly from the predictions of the conventional Fermi liquid picture and that the ground state of Al is modified anomalously by the addition of Li. PMID- 16712382 TI - Evidence for two-path recombination of photoinduced small polarons in reduced LiNbO(3). AB - The recombination of photoinduced free Nb(4+)(Nb) and bound Nb(4+)(Li) small polarons toNb(4+)(Li): Nb(4+)(Nb) bipolarons is investigated in nominally pure, reduced LiNbO(3) single crystals by means of excited-state-absorption spectroscopy. We discovered a two-component decay of the light-induced absorption alpha(li)(t) for probe light at lambda=785 nm and moderate pump beam intensities (I(p) << 670 GW/m(2)). These experimental results give strong evidence for the existence of a two-path recombination of the photoinduced polarons. A corresponding model taking into account hopping charge transport and trapping is presented. PMID- 16712383 TI - Decomposition of 1/f noise in Al(x)Ga(1-x)As/GaAs Hall devices. AB - We present a systematic study of the low-frequency noise in micron and submicron Hall devices made from Al(x)Ga(1-x)As/GaAs heterostructures. In a sample with feature size as small as 0.45 microm we observe a nonmonotonic temperature dependence of the noise power spectral densities (PSD's) at temperatures where surface states and deep-level excitations are frozen out. Near the temperature where the noise peaks, the PSD's can be described by a thermally activated two level random telegraph signal, i.e., the 1/f noise originating from switching events in the highly doped Al(x)Ga(1-x) layer is resolved into a single Lorentzian spectrum. PMID- 16712384 TI - Theory of phonon-induced spin relaxation in laterally coupled quantum dots. AB - Phonon-induced spin relaxation in coupled lateral quantum dots in the presence of spin-orbit coupling is calculated. The calculation for single dots is consistent with experiment. Spin relaxation in double dots at useful interdot couplings is dominated by spin-hot spots that are strongly anisotropic. Spin-hot spots are ineffective for a diagonal crystallographic orientation of the dots with a transverse in-plane field. This geometry is proposed for spin-based quantum information processing. PMID- 16712385 TI - Spin torque, tunnel-current spin polarization, and magnetoresistance in MgO magnetic tunnel junctions. AB - We employ the spin-torque response of magnetic tunnel junctions with ultrathin MgO tunnel barrier layers to investigate the relationship between spin transfer and tunnel magnetoresistance (TMR) under finite bias, and find that the spin torque per unit current exerted on the free layer decreases by < 10% over a bias range where the TMR decreases by > 40%. This is inconsistent with free-electron like spin-polarized tunneling and reduced-surface-magnetism models of the TMR bias dependence, but is consistent with magnetic-state-dependent decay lengths in the tunnel barrier. PMID- 16712386 TI - Domain wall creation in nanostructures driven by a spin-polarized current. AB - We report on current-driven magnetization reversal in nanopillars with elements having perpendicular magnetic anisotropy. Whereas only the two uniform magnetization states are available under the action of a magnetic field, we observed current-induced Bloch domain walls in pillars as small as 50 x 100 nm(2). This domain wall state can be further controlled by current to restore the uniform states. The ability to nucleate and manipulate domain walls by a current gives insight into the reversal mechanisms of small nanoelements and provides new prospects for ultrahigh density spintronic devices. PMID- 16712387 TI - Spectral dependence of spin photocurrent and current-induced spin polarization in an InGaAs/InAlAs two-dimensional electron gas. AB - The converse effects of spin photocurrent and current induced spin polarization are experimentally demonstrated in a two-dimensional electron gas system with Rashba spin splitting. Their consistency with the strength of the Rashba coupling as measured for the same system from beating of the Shubnikov-de Haas oscillations reveals a unified picture for the spin photocurrent, current-induced spin-polarization, and spin-orbit coupling. In addition, the observed spectral inversion of the spin photocurrent indicates a system with dominating structure inversion asymmetry. PMID- 16712388 TI - Alkali-induced enhancement of surface electronic polarizibility. AB - From results of ab initio electronic structure calculations based on density functional theory for a set of prototype systems, we find alkali adsorbates to cause a dramatic enhancement of the electronic polarizability of the metal surface extending it several angstroms into the vacuum. This phenomenon is traceable to an unusual feature induced in the surface potential on alkali adsorption. The effect appears to be general, as we find it to be present on metals as varied as Pd and Cu, and helps explain the observed substantial decrease in the vibrational frequency of molecules when coadsorbed with alkalis on metal surfaces. Specifically, for two dissimilar molecules CO and O(2), we trace the softening of the frequencies of their stretching mode when coadsorbed with K on Pd(111) to the enhanced electronic polarizability. PMID- 16712389 TI - Quantum criticality of a Fermi gas with a spherical dispersion minimum. AB - We describe the quantum phase transition of a Fermi gas occurring when the quasiparticle excitation energy has a minimum in momentum space which crosses zero on a sphere of radius k(0) not equal 0. The quasiparticles have a universal interaction which controls the physical properties in the vicinity of the quantum critical point. We discuss possible applications to fermionic superfluids formed by pairing two fermion species, near the point where the densities of the two species become unequal. PMID- 16712390 TI - Continuous-time monitoring of Landau-Zener interference in a cooper-pair box. AB - Landau-Zener (LZ) tunneling can occur with a certain probability when crossing energy levels of a quantum two-level system are swept across the minimum energy separation. Here we present experimental evidence of quantum interference effects in solid-state LZ tunneling. We used a Cooper-pair box qubit where the LZ tunneling occurs at the charge degeneracy. By employing a weak nondemolition monitoring, we observe interference between consecutive LZ-tunneling events; we find that the average level occupancies depend on the dynamical phase. The system's unusually strong linear response is explained by interband relaxation. Our interferometer can be used as a high-resolution Mach-Zehnder-type detector for phase and charge. PMID- 16712391 TI - Entanglement and dynamics of spin chains in periodically pulsed magnetic fields: accelerator modes. AB - We study the dynamics of a single excitation in a Heisenberg spin-chain subjected to a sequence of periodic pulses from an external, parabolic, magnetic field. We show that, for experimentally reasonable parameters, a pair of counterpropagating coherent states is ejected from the center of the chain. We find an illuminating correspondence with the quantum time evolution of the well-known paradigm of quantum chaos, the quantum kicked rotor. From this we can analyze the entanglement production and interpret the ejected coherent states as a manifestation of the so-called "accelerator modes" of a classically chaotic system. PMID- 16712392 TI - Observation of spin-wave soliton fractals in magnetic film active feedback rings. AB - The manifestation of fractals in soliton dynamics has been observed for the first time. The experiment utilized self-generated spin-wave envelope solitons in a magnetic film based active feedback ring. At high ring gain, the soliton that circulates in the ring breathes in a fractal pattern. The corresponding power frequency spectrum shows a comb structure, with each peak in the comb having its own comb, and so on, to finer and finer scales. PMID- 16712393 TI - Time-domain ab initio simulation of electron and hole relaxation dynamics in a single-wall semiconducting carbon nanotube. AB - The electron and hole relaxation in the (7, 0) zigzag carbon nanotube is simulated in time domain using a surface-hopping Kohn-Sham density functional theory. Following a photoexcitation between the second van Hove singularities, the electrons and holes decay to the Fermi level on characteristic subpicosecond time scales. Surprisingly, despite a lower density of states, the electrons relax faster than the holes. The relaxation is primarily mediated by the high-frequency longitudinal optical (LO) phonons. Hole dynamics are more complex than the electron dynamics: in addition to the LO phonons, holes couple to lower frequency breathing modes and decay over multiple time scales. PMID- 16712394 TI - Nonlinear terahertz response of -type GaAs. AB - Excitation of an n-type GaAs layer by intense ultrashort terahertz pulses causes coherent emission at 2 THz. Phase-resolved nonlinear propagation experiments show a picosecond decay of the emitted field, despite the ultrafast carrier-carrier scattering at a sample temperature of 300 K. While the linear THz response is in agreement with the Drude response of free electrons, the nonlinear response is dominated by the super-radiant decay of optically inverted impurity transitions. A quantum mechanical discrete state model using the potential of the disordered impurities accounts for all experimental observations. PMID- 16712395 TI - Nanosecond domain wall dynamics in ferroelectric Pb(Zr, Ti)O(3) thin films. AB - Domain wall motion during polarization switching in ferroelectric thin films is fundamentally important and poses challenges for both experiments and modeling. We have visualized the switching of a Pb(Zr, Ti)O(3) capacitor using time resolved x-ray microdiffraction. The structural signatures of switching include a reversal in the sign of the piezoelectric coefficient and a change in the intensity of x-ray reflections. The propagation of polarization domain walls is highly reproducible from cycle to cycle of the electric field. Domain wall velocities of 40 m s(-1) are consistent with the results of other methods, but are far less than saturation values expected at high electric fields. PMID- 16712396 TI - Importance of second-order piezoelectric effects in zinc-blende semiconductors. AB - We show that the piezoelectric effect that describes the emergence of an electric field in response to a crystal deformation in III-V semiconductors such as GaAs and InAs has strong contributions from second-order effects that have been neglected so far. We calculate the second-order piezoelectric tensors using density-functional theory and obtain the piezoelectric field for [111]-oriented In(x)Ga(1-x)As quantum wells of realistic dimensions and concentration x. We find that the linear and the quadratic piezoelectric coefficients have the opposite effect on the field, and for large strains (large In concentration) the quadratic terms even dominate. Thus, the piezoelectric field turns out to be a rare example of a physical quantity for which the first-order and second-order contributions are of comparable magnitude. PMID- 16712397 TI - Decoupling phenomena in supercooled liquids: signatures in the energy landscape. AB - A significant deviation from the Debye model of rotational diffusion in the dynamics of orientational degrees of freedom in an equimolar mixture of ellipsoids of revolution and spheres is found to begin at a temperature at which the average inherent structure energy of the system starts falling with drop in temperature. We argue that this onset temperature corresponds to the emergence of the process as a distinct mode of orientational relaxation. Further, we find that the coupling between rotational and translational diffusion breaks down at a still lower temperature where a change occurs in the temperature dependence of the average inherent structure energy. PMID- 16712398 TI - Collapse of telechelic star polymers to watermelon structures. AB - Conformational properties of star-shaped polymer aggregates that carry attractive end groups, called telechelic star polymers, are investigated by simulation and analytical variational theory. We focus on the case of low telechelic star polymer functionalities, f < or = 5, a condition which allows aggregation of all attractive monomers on one site. We establish the functionality- and polymerization-number dependence of the transition temperature from the "star burst" to the "watermelon" macroparticle structure. Extensions to telechelic stars featuring partially collapsed configurations are also discussed. PMID- 16712399 TI - Genetic toggle switch without cooperative binding. AB - Genetic switch systems with mutual repression of two transcription factors are studied using deterministic and stochastic methods. Numerous studies have concluded that cooperative binding is a necessary condition for the emergence of bistability in these systems. Here we show that, for a range of biologically relevant conditions, a suitable combination of network structure and stochastic effects gives rise to bistability even without cooperative binding. PMID- 16712400 TI - Mechanically controlled DNA extrusion from a palindromic sequence by single molecule micromanipulation. AB - A magnetic tweezers setup is used to control both the stretching force and the relative linking number DeltaLk of a palindromic DNA molecule. We show here, in absence of divalent ions, that twisting negatively the molecule while stretching it at approximately 1 pN induces the formation of a cruciform DNA structure. Furthermore, once the cruciform DNA structure is formed, the extrusion of several kilo-base pairs of palindromic DNA sequence is directly and reversibly controlled by varying DeltaLk. Indeed the branch point behaves as a nanomechanical gear that links rotation with translation, a feature related to the helicity of DNA. We obtain experimentally a very good linear relationship between the extension of the molecule and DeltaLk. We use then this experiment to obtain a precise measurement of the pitch of B-DNA in solution: 3.61 +/- 0.03 nm/turn. PMID- 16712401 TI - Dynamics and pattern formation in invasive tumor growth. AB - We study the in vitro dynamics of the malignant brain tumor glioblastoma multiforme. The growing tumor consists of a dense proliferating zone and an outer less dense invasive region. Experiments with different types of cells show qualitatively different behavior: one cell line invades in a spherically symmetric manner, but another gives rise to branches. We formulate a model for this sort of growth using two coupled reaction-diffusion equations for the cell and nutrient concentrations. When the ratio of the nutrient and cell diffusion coefficients exceeds some critical value, the plane propagating front becomes unstable with respect to transversal perturbations. The instability threshold and the full phase-plane diagram in the parameter space are determined. The results are in a qualitative agreement with experimental findings for the two types of cells. PMID- 16712402 TI - Evolutionary dynamics on degree-heterogeneous graphs. AB - The evolution of two species with different fitness is investigated on degree heterogeneous graphs. The population evolves either by one individual dying and being replaced by the offspring of a random neighbor (voter model dynamics) or by an individual giving birth to an offspring that takes over a random neighbor node (invasion process dynamics). The fixation probability for one species to take over a population of N individuals depends crucially on the dynamics and on the local environment. Starting with a single fitter mutant at a node of degree k, the fixation probability is proportional to k for voter model dynamics and to 1/k for invasion process dynamics. PMID- 16712403 TI - Dynamics, rectification, and fractionation for colloids on flashing substrates. AB - We show that a rich variety of dynamic phases can be realized for mono- and bidisperse mixtures of interacting colloids under the influence of a symmetric flashing periodic substrate. With the addition of dc or ac drives, phase locking, jamming, and new types of ratchet effects occur. In some regimes we find that the addition of a nonratcheting species increases the velocity of the ratcheting particles. We show that these effects occur due to the collective interactions of the colloids. PMID- 16712404 TI - Star polymers in shear flow. AB - Linear and star polymers in solution are studied in the presence of shear flow. The solvent is described by a particle-based mesoscopic simulation technique, which accounts for hydrodynamic interactions. The scaling properties of the average gyration tensor, the orientation angle, and the rotation frequency are investigated for various arm lengths and arm numbers. With increasing functionality f, star polymers exhibit a crossover in their flow properties from those of linear polymers to a novel behavior, which resembles the tank-treading motion of elastic capsules. PMID- 16712405 TI - Classification scheme for phenomenological universalities in growth problems in physics and other sciences. AB - A classification in universality classes of broad categories of phenomenologies, belonging to physics and other disciplines, may be very useful for a cross fertilization among them and for the purpose of pattern recognition and interpretation of experimental data. We present here a simple scheme for the classification of nonlinear growth problems. The success of the scheme in predicting and characterizing the well known Gompertz, West, and logistic models, suggests to us the study of a hitherto unexplored class of nonlinear growth problems. PMID- 16712406 TI - Comment on "Gaussian quantum Monte Carlo methods for fermions and bosons". PMID- 16712408 TI - Comment on "Theory of current-driven domain wall motion: spin transfer versus momentum transfer". PMID- 16712410 TI - Comment on "Bose-Einstein condensation of magnons in Cs(2)CuCl(4)". PMID- 16712412 TI - Comment on "Theoretical analysis of the transmission phase shift of a quantum dot in the presence of Kondo correlations". PMID- 16712415 TI - An interview with Wayne Bowen, Ph.D. Interview by Vicki Glaser. PMID- 16712417 TI - A high-content screening assay for the Nogo receptor based on cellular Rho activation. AB - Rho family proteins can coordinate multiple signaling pathways through their ability to regulate both gene transcription and the actin cytoskeleton. With respect to the neuronal Nogo receptor (NgR), recent data assign a key role for the GTPase Rho in the control of cellular responses leading to actin cytoskeletal rearrangements and finally resulting in axonal outgrowth inhibition and growth cone collapse in the adult human central nervous system. In order to evaluate potential NgR antagonists, human embryonic kidney 293 cells stably overexpressing RhoA in the absence or presence of NgR have been generated. RhoA activation induced by stimulation with the alkaline phosphatase-tagged NgR ligand Nogo66 (AP Nogo66) was confirmed by affinity-precipitation of the GTPase with the Rho binding domain from Rhotekin. As this pull-down assay is not applicable to a higher-throughput format, a cellular Rho GTPase activation assay strategy based on the ability of Rho to regulate the actin cytoskeleton was developed. Stimulation with L-alpha-lysophosphatidic acid (LPA), a Rho activator acting through the ubiquitiously expressed LPA receptors, induced significant cytoskeletal rearrangement resulting in cell contraction in all RhoA overexpressing cell lines. In contrast, stimulation with AP-Nogo66 resulted in Rho-dependent cell contraction with a similar time course only in the NgR expressing cell line. Moreover, the NgR-induced Rho-dependent morphological changes could be analyzed and quantified with customary image analysis software. In conclusion, the cytoskeletal rearrangement assay is amenable to automated high content screening and has the potential to eliminate major technical bottlenecks of the pull-down assay. The increased throughput of the cellular GTPase activation assay compared with the biochemical method should facilitate the evaluation of compounds that modulate the actin cytoskeleton through Rho. PMID- 16712418 TI - A morphology- and kinetics-based cascade for human neural cell high content screening. AB - The prospect of manipulating endogenous neural stem cells to replace damaged tissue and correct functional deficits represents a novel mechanism for treating a variety of central nervous system disorders. Using human neural precursor cultures and a variety of assays for studying stem cell behavior we have screened two libraries of commercially available compounds using an endpoint high content screening assay. We then performed detailed follow-up mechanistic studies on confirmed hits using endpoint and kinetics assays to characterize and differentiate the mechanisms of action of these compounds. The screening cascade employed successfully identified a number of active compounds with differing mechanisms of action. This approach shows how hits from a phenotypic screen can be prioritized and characterized by high content screening to identify potentially novel mechanisms and druggable targets to take forward into more conventional high-throughput screening approaches. PMID- 16712419 TI - The use of high-content screening for the discovery and characterization of compounds that modulate mitotic index and cell cycle progression by differing mechanisms of action. AB - The advent of high-content screening has expanded the ability of researchers to identify and quantify compound effects on a number of cellular events in a manner that allows for the rapid screening of chemical libraries. We have validated an approach for characterizing inhibitors of Aurora kinase family members using high content screening by determining compound effects on the levels of the mitotic marker phospho-histone H3 (Ser10). Analysis of the data from these experiments led us to the discovery of a series of related compounds that increased the level of cells staining positive for the mitotic marker, indicating a block of cell cycle progression at M-phase. We then reconfigured the Aurora kinase inhibition assay and validated a high-content approach to identify compounds that block progression through M-phase. We were able to take advantage of the flexibility within the high-content screening platform to measure DNA content, activation of apoptosis, and effects on beta-tubulin staining patterns, all of which directly led to the identification of the cellular target of this new class of compounds. PMID- 16712420 TI - An innovative cell-based assay for the detection of modulators of soluble guanylate cyclase. AB - Guanylate cyclase (GC) catalyzes the biosynthesis of cyclic guanosine 3',5'- monophosphate (cGMP) from GTP. GC exists in two isoenzyme forms: soluble and membrane-bound. The soluble GC (sGC) is a heterodimer composed of an alpha and a beta subunit, and it contains heme as a prosthetic group. The most important physiological activator of sGC is nitric oxide, which activates the enzyme upon binding to the heme moiety. By producing the second messenger cGMP, which regulates various effector systems such as phosphodiesterases, ion channels, and protein kinases, sGC plays an important role in different physiological processes, thus representing a very attractive pharmacological target. In fact, the pathogenesis of several diseases, especially those of the cardiovascular system, has been linked to inappropriate regulation of sGC. In order to find new modulators for this important enzyme, an innovative cell-based assay has been developed and optimized for the use in high-throughput screening. This luminescent assay, which is suitable for both 96- and 384-well plate formats, has been achieved by stably expressing the alpha and beta subunits of a mutated form of sGC in Chinese hamster ovary cells. The mutated form synthesizes cyclic adenosine 3',5'-monophosphate instead of cGMP, allowing the detection of enzymatic activity by a reporter gene approach. We demonstrated that this cell line responds to compounds typically used in the field of sGC research and that it represents an innovative and robust assay to screen for sGC modulators with high efficiency and high sensitivity by means of standard luminescence readers. PMID- 16712421 TI - A high-throughput microfluidic assay for SH2 domain-containing inositol 5 phosphatase 2. AB - SH2 domain-containing inositol 5-phosphatase 2 (SHIP2) is a potential drug target for the treatment of type 2 diabetes. This enzyme serves as a negative regulator of insulin-mediated signal transduction by catalyzing the dephosphorylation of the second messenger lipid molecule phosphatidylinositol 3,4,5-triphosphate. Traditionally, assays for phosphoinositide phosphatases such as SHIP2 have relied on radiolabeled phosphatidylinositol-containing lipid membranes and chromatographic separation of labeled phospholipid substrate from product by thin layer chromatography. We have expressed and purified catalytically active phosphatase domain constructs of SHIP2 from Escherichia coli and developed a sensitive and antibody- or binding protein-independent assay for SHIP2 amenable to high-throughput screening of phosphoinositide phosphatases or phosphoinositide kinases. This microfluidic assay, with Z' values approximately 0.8, is based upon the difference in mobility within an electric field between a fluorophore-labeled phosphatidylinositol 3,4,5-triphosphate substrate and the corresponding 3,4 bisphosphate product. High-throughput screening of a 91,060-member compound library in 384-well format resulted in the identification of SHIP2 inhibitors. PMID- 16712422 TI - Overcoming the hurdle of fluorescent compounds in kinase screening: a case study. AB - In the field of screening in general and especially in the kinase area, taking into consideration throughput and cost, fluorescence- and luminescence-based assays have been developed as alternatives to radioactivity-based assays. However, fluorescence-based technologies are not devoid of pitfalls. One of the main problems is interference from autofluorescent compounds and the incidence of false-positives as exemplified here with a fluorescence polarization (FP)-based assay. Using the scintillation proximity assay as the in-house standard, we assessed several alternatives to radioactive methods, namely, the amplified luminescent proximity homogeneous assay screen (ALPHAScreen, Perkin-Elmer Life Sciences, Boston, MA), enzyme fragment complementation, FP, and nanofluidics based fluorescence intensity. Data comparing the sensitivity, robustness, relative sensitivity to autofluorescent compounds, enzyme consumption, and relative costs of each assay for one common kinase are presented. Results obtained seem to favor the nanofluidics mobility shift assay as a method of choice, followed by the direct FP approach, using generic high-molecular-weight phosphate group-binders. PMID- 16712423 TI - Performance characteristics of colorimetric protein microarrays compared to ELISA. AB - Miniaturization and multiplexing are two areas that need improvement in immunoassays. However, it is unclear how associated changes in size, substrate, binding capacity, protein concentration, and other parameters affect sensitivity and quantitative range. In the study here we compared a new protein microarray substrate and system (Zeta-Grip, Miragene Inc., Anaheim, CA) with enzyme-linked immunosorbent assay (ELISA) for the application of immunoassays. We found that the protein microarray system performed better than ELISA in both sensitivity and quantitative range for use in immunoassay. PMID- 16712424 TI - Evaluation of MTT assay for measurement of emodin-induced cytotoxicity. AB - The specificity and sensitivity of the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) tetrazolium assay can be influenced by certain factors, such as colored substances and cell volume. When the MTT assay is applied to measure cytotoxicity induced by emodin, its accuracy might be affected by emodin itself. Vascular smooth muscle cells were cultured in M199 medium. The optical density of emodin or formazan was measured by spectrophotometry. Emodin has a different absorption spectrum in different solvents. The solvents containing water induced a red shift of the absorption curve of emodin, which increased the overlap of the absorption curves of emodin and formazan. Formazan was formed from the MTT tetrazolium salt by emodin in a dose-dependent manner, which was partially suppressed by serum. Cytotoxicity was induced by emodin in a time- and dose-dependent manner in a modified MTT assay. The data suggest that emodin can alter the accuracy of the MTT assay but that a modified MTT assay is still valuable in measuring emodin-induced cytotoxicity. PMID- 16712425 TI - Application of laser-scanning fluorescence microplate cytometry in high content screening. AB - The resolution of cell-based assays down to the cellular level has created new opportunities for the drug discovery process. Aptly named high content analysis, such an approach is enabling new methods of analysis for the broad range of therapeutic targets emerging in the post-genomics era, and offering alternative multiparametric readouts for some traditional analyses. Microplate cytometry is one of the technologies that is being applied to a broad range of assays utilizing fluorescent labeling, at throughputs compatible with primary screening campaigns. Cellular resolution is achieved using scanning laser excitation coupled to photomultiplier detection. This configuration results in area-based scanning across a large field of view, plus simultaneous detection of up to four emission colors for efficient multiplexing. Microplate cytometry is being used most extensively in the field of oncology research because of its usefulness for numerous applications, including protein kinase activity, cell cycle analysis, and cell colony formation. The review focuses on the Acumen Explorer microplate cytometer (TTP LabTech Ltd., Melbourn, Hertfordshire, UK), detailing the principal components of the instrument and providing an overview of its use in high content screening. PMID- 16712427 TI - Predictors of health status in women with fibromyalgia: a prospective study. AB - Although cross-sectional studies have identified correlates of dysfunction in fibromyalgia (FM) patients (e.g., psychological distress and pain), predictors of health status have not been previously investigated using a longitudinal research design. We gathered data from 156 women who met American College of Rheumatology criteria for primary FM recruited from both tertiary care and community settings. Stepwise multiple linear regression analysis indicated that poorer health status (p < .0001) and more comorbidity (p = .0089) at baseline were predictors of poorer health status 6 months later. After controlling for these covariates, psychological distress contributed significantly to the model (p = .01). There was a trend indicating that palliative coping styles (i.e., self-care, energy conservation) altered the impact of pain on 6-month health status (p = .06). These findings highlight the need for multidisciplinary interventions that target psychological distress, coping, and comorbidity in patients with FM. PMID- 16712429 TI - School and leisure activities in adolescents and young adults with chronic digestive disorders: impact of burden of disease. AB - First, we compared the nature of burden of disease (i.e., manifestations of the disease in daily life) in adolescents and young adults with various chronic digestive disorders with controls. After that, we investigated whether burden of disease is associated with difficulties in school and leisure activities of adolescents and young adults with various digestive disorders. For this purpose, we performed a multicenter study in 5 diagnostic groups (total N = 758; ages 12 to 25 years) including inflammatory bowel diseases (IBD), chronic liver diseases, congenital disorders, celiac disease, and food allergy and a population based control group (N = 306) using a self-report questionnaire. Especially adolescents and young adults with a chronic liver disease, IBD, and food allergy were found to experience daily manifestations of their disease. Several disease burden characteristics, of which especially depression, could be identified as important contributors to difficulties in school performance and leisure activities. PMID- 16712428 TI - Violence exposure and cortisol responses in urban youth. AB - I examined the physiological costs associated with exposure to violence in 101 African American youth (55% male; M age = 11.14 years) living in high-violence areas of a midsized southern city in the United States. Salivary cortisol was measured before and after a laboratory task (viewing and discussing a video depicting community violence) and on waking 1 morning in the week following the laboratory assessment. Overall, cortisol levels were low. Analyses controlling for age, gender, negative affect, and major life events revealed that peer victimization was associated with lower basal cortisol values obtained from home assessments. Witnessed violence predicted a cortisol awakening response (CAR) but only in girls; girls with a typical CAR had lower levels of witnessing violence than girls with an atypical pattern. Witnessed violence also was associated with lower baseline cortisol levels measured in the laboratory and with increases in cortisol from baseline to posttask for boys but not girls. Peer victimization was associated with increases in cortisol from pretask to posttask for both genders. I discuss implications for research and prevention. PMID- 16712430 TI - Total workload as related to psychological well-being and symptoms in full-time employed female and male white-collar workers. AB - Most research on the combined effects of paid and unpaid workload has related these factors to stress, depression, and physical symptoms. Thus, comparative knowledge concerning positive aspects of human functioning, such as health and well-being and how they relate to total workload of employed women and men, is limited. Our aim in this study was to investigate how total workload including paid and unpaid work is related to psychological well-being and symptoms in full time employed women and men. We obtained data on workload, general symptoms, and the Ryff scales covering self-acceptance, environmental mastery, positive relations, personal growth, purpose in life, and autonomy from questionnaires mailed to a stratified sample of highly educated white-collar workers aged between 32 and 58 years. Data from women (n = 430) and men (n = 400) living in partner relationships with at least one child showed that increasing hours of unpaid work was associated with decreasing levels of self-acceptance and environmental mastery in women, whereas paid work was associated with increasing levels of personal growth and decreasing levels of purpose in life. For men, paid work was associated with increasing levels of personal growth and more symptoms. We discuss factors underlying the gender-specific relationships between paid and unpaid work, psychological well-being, and symptoms. PMID- 16712431 TI - Trait anger and blood pressure recovery following acute pain: evidence for opioid mediated effects. AB - Previous work has suggested that positive associations between trait anger (TRANG) and pain sensitivity are due to dysfunctional endogenous opioid analgesic systems. In this study, we examined whether TRANG is associated with impaired opioid modulation of blood pressure (BP) recovery. A total of 46 pain-free normotensive controls and 69 normotensive chronic low back pain (LBP) sufferers received opioid blockade (8 mg naloxone i.v.) or placebo in randomized, counterbalanced order in separate sessions. During each, participants underwent a 1-min finger pressure pain task followed by an ischemic forearm pain task. Opioid blockade impaired post-pain BP recovery in controls but not LBP participants (ps < .001). In controls, low TRANG was associated with blockade-induced recovery impairments, with no blockade effect in high TRANG participants. In LBP participants, blockade did not alter recovery regardless of TRANG (interaction ps < .05). Results support dysfunctional opioid modulation of BP recovery in healthy high TRANG controls and further suggest chronic pain-related impairments in opioid-mediated cardiovascular recovery. PMID- 16712432 TI - Dimensions of hostility in Japanese undergraduate students. AB - Japanese undergraduate students (N = 512) completed the Japanese versions of 4 hostility questionnaires: Aggression Questionnaire (Ando et al., 1999); State Trait Anger Expression Inventory (Suzuki & Haruki, 1994); Muller Anger Coping Questionnaire (Otake et al., 2000); and Cynicism Questionnaire (Izawa & Nomura, 2004). Dimensions of hostility, determined by factor analysis for the 12 subscales of the 4 questionnaires, revealed a 4-factor solution: Expressive Hostility, Inhibitive Hostility, Assertiveness, and Hostile Cognition. Prior studies with Western participants had suggested a 2-factor solution of Expression and Experience dimensions of hostility. However, our results forcing a 2-factor solution showed a different pattern: The first factor included Expressive Hostility and Hostile Cognition, and the second factor included Inhibitive Hostility. These results indicate that the dimensions of hostility in Japanese students differed from those in American and European participants. This may be related to characteristics of Japanese society that emphasize cooperative behavior and not expressing angry emotions. PMID- 16712433 TI - Sociodemographic correlates of selected health risk behaviors in a representative sample of Australian young people. AB - To facilitate the development of interventions to reduce health risk behavior among young people, we designed this study to compare risk behavior among young people and older people, to compare risk behavior profiles between young men and women, and to identify sociodemographic correlates of risk behavior among young people. Computer-assisted telephone interviews with a representative sample of 19,307 Australian men and women (response rate 73.1%) assessed alcohol consumption, cigarette smoking, injection drug use, and unprotected intercourse. Respondents aged 16 to 24 reported less healthy behavior than older people. Although men and women aged 16 to 24 had similar profiles of health risk behavior, correlates of these behaviors differed for men and women. There were few consistent sociodemographic correlates of different risk behaviors. The results suggest that young women are now as important a priority as young men for interventions. Young people remain an important target group for health promotion, with nonheterosexual young people a particular high-risk group. PMID- 16712434 TI - Association between hot flashes, sleep complaints, and psychological functioning among healthy menopausal women. AB - Self-report data suggest that sleep hot flashes among menopausal women are associated with sleep problems and in turn impaired psychological functioning. However, few studies have examined these relations with physiologic hot flash measures. A total of 41 perimenopausal and postmenopausal women with daily hot flashes underwent nighttime sternal skin conductance monitoring to quantify hot flashes. Participants completed sleep diaries; the Sleep-Wake Experience List (van Diest, 1990); and depression, anxiety, and daily stress measures. Participants experienced a median of 2 physiologically monitored and 1 reported sleep hot flash nightly. Although sleep complaints were significantly and positively associated with psychological functioning, neither sleep complaints nor psychological functioning was significantly related to frequency of physiologically monitored sleep hot flashes. Conversely, results indicate an association between reported sleep hot flashes and acute sleep problems. The frequency of physiologically monitored sleep hot flashes, as opposed to reported sleep hot flashes, may be independent of problems with sleep and mood among menopausal women. PMID- 16712435 TI - Relations of caregiving stress and health depend on the health indicators used and gender. AB - Extensive research has evaluated relations between stress and health. These studies have varied in the type of stress examined (acute vs. chronic) and in the way in which health has been operationalized. Here we examine relations between chronic stress and 25 indicators of various health dimensions (e.g., physiological indexes, medical records, and self-reports of global health; symptoms, functional status, health service utilization, and psychosocial distress/quality of life). We also assessed whether such relations are moderated by gender, an individual difference variable that is important to health and longevity. Samples included 157 community-residing older adults (M age = 69.4 years, 31.8% men), approximately half of whom were caregivers for a spouse with Alzheimer's disease, and half were demographically similar noncaregiver spouses. Principal component analyses on the 25 health measures resulted in 5 factors that met standard criteria for acceptance. In women, caregivers reported worse physical health and psychological health than noncaregivers, but their physiological risk was similar. In men, caregivers had greater physiological risk, but they reported better physical health than did men noncaregivers. Researchers who study chronic stress and health should consider the possibility that the relation between chronic stress and health may vary for men and women depending on the type of health being assessed. PMID- 16712436 TI - Biochemical aspects of coronavirus replication and virus-host interaction. AB - Infection by different coronaviruses (CoVs) causes alterations in the transcriptional and translational patterns, cell cycle, cytoskeleton, and apoptosis pathways of the host cells. In addition, CoV infection may cause inflammation, alter immune and stress responses, and modify the coagulation pathways. The balance between the up- and downregulated genes could explain the pathogenesis caused by these viruses. We review specific aspects of CoV-host interactions. CoV genome replication takes place in the cytoplasm in a membrane protected microenvironment and may control the cell machinery by locating some of their proteins in the host cell nucleus. CoVs initiate translation by cap dependent and cap-independent mechanisms. CoV transcription involves a discontinuous RNA synthesis (template switching) during the extension of a negative copy of the subgenomic mRNAs. The requirement for base-pairing during transcription has been formally demonstrated in arteriviruses and CoVs. CoV N proteins have RNA chaperone activity that may help initiate template switching. Both viral and cellular proteins are required for replication and transcription, and the role of selected proteins is addressed. PMID- 16712437 TI - Ways to say no: refusal skill strategies among urban adolescents. AB - OBJECTIVES: To examine associations among adolescents' generated verbal strategies (ie, Simple No, Declarative Statements, Excuse, Alternatives) and underlying nonverbal assertiveness in 2 refusal situations: smoking and shoplifting. METHODS: Sixth-grade urban minority students (N = 454) participated in videotaped role-play assessments of peer refusal skills. RESULTS: Differences were found by situation with students demonstrating greater use of Simple No in the smoking refusal and Alternatives in the shoplifting refusal. Nonverbal assertiveness was similar across situations and was associated with Declarative Statements, but only in the smoking refusal. CONCLUSIONS: Prevention programs should tailor refusal skills practice to cover multiple situations. PMID- 16712438 TI - Heterogeneity in the smoking behavior of African American women. AB - OBJECTIVE: To assess the association between sociodemographic variables and smoking behavior patterns of African American women. METHODS: Six years of data (N=14,903) from the National Health Interview Surveys were analyzed using multiple logistic regression. RESULTS: African American women in the South were more likely to never smoke and to start smoking later than women in the Northeast. Positive smoking outcomes (never smoking, initiating smoking at later ages, and quitting) were associated with higher education, higher income, and being married. CONCLUSIONS: Variations among African American women suggest the need for targeting specific subgroups at greater risks to reduce disparities in smoking and smoking-related diseases. PMID- 16712440 TI - Burn injuries among infants and toddlers in the United States, 1997-2002. AB - OBJECTIVES: To determine the magnitude and causes of unintentional burn injuries attributed to hot objects or substances among children aged birth to 4 years in the United States. METHODS: Data were obtained from the National Hospital Ambulatory Medical Care System. RESULTS: Annually, 78,000 infants and toddlers were treated in ambulatory care settings for injuries due to contact with a hot object or substance. Hot foods, curling irons, or clothing irons were the main causes of burns. CONCLUSIONS: Due to the varied causes of burns, a combination of health education and product design interventions are likely necessary to prevent these injuries. PMID- 16712439 TI - Age of smoking onset as a predictor of smoking cessation during pregnancy. AB - OBJECTIVE: To assess age of smoking onset as a predictor of smoking cessation during pregnancy. METHODS: Multivariate logistic regression analysis of cross sectional data for 35,687 women 15-34 years old from the 2000 and 2001 National Household Survey on Drug Abuse. RESULTS: Pregnant women (1805) were twice as likely to abstain as nonpregnant women (47.7% vs 23.3%). Initiation of smoking at >15 years old was associated with increased likelihood of 30-day abstinence during pregnancy (OR=1.75) after adjusting for confounders. CONCLUSIONS: The earlier a woman starts smoking, the more likely it is that she will smoke during pregnancy, underscoring the need for early prevention to protect mothers and the fetuses. PMID- 16712441 TI - Does the type of exposure to a roadside sobriety checkpoint influence driver perceptions regarding drunk driving? AB - OBJECTIVES: To determine if the type of exposure to roadside sobriety checkpoints is related to vulnerability of being stopped by the police for drinking and driving. METHODS: Random household telephone surveys were conducted over 3 years. RESULTS: Personally exposed drivers had lower perceptions of vulnerability than did drivers who knew a family member who had been stopped. Exposed drivers who also had a family member and an acquaintance that had been stopped had higher perceptions of vulnerability. CONCLUSIONS: Direct exposure to a checkpoint appears to lower perceptions of risk than if one has had only indirect exposure (ie, family or friends). PMID- 16712442 TI - Social support and youth physical activity: the role of provider and type. AB - OBJECTIVE: To examine provider and type variation in social support (SS) for activity. METHODS: Three hundred sixty-three fifth to eighth-grade students completed a questionnaire assessing self-reported activity and social support (SS) from 3 providers: mom, dad, and peers. Important covariates of activity were included in the analysis: age, BMI, sex, and maturation. RESULTS: Structural equation modeling indicated peers, transportation, and praise affected activity levels. Boys reported greater SS than girls did. Maturation, age, and BMI exhibited unique affects on SS. CONCLUSIONS: Increasing positive feedback, transportation to places to be active, and peer support may prove advantageous in improving activity levels in this age-group. PMID- 16712443 TI - Stages of change in physical exercise: a test of stage discrimination and nonlinearity. AB - OBJECTIVES: To investigate the utility of the protection motivation theory (PMT), social-cognitive theory (SCT) and transtheoretical model's (TTM) processes of change (POC) by (1) discriminating between TTM's stages with planned comparisons and (2) examining higher-ordered trends across stages. METHODS: 1582 individuals were included in the analyses that tested severity, vulnerability, response efficacy, self-efficacy, POC, pros and cons, social and environmental support. RESULTS: The findings provide evidence for the utility of the PMT and TTM's POC for stage discrimination; stage discontinuity patterns are supported. CONCLUSIONS: Promotion of health behavior should target stage-specific variables, such as threat appraisals for early stage movements. PMID- 16712444 TI - Young adult smoker risk perceptions of traditional cigarettes and nontraditional tobacco products. AB - OBJECTIVE: To explore risk perceptions of traditional and nontraditional tobacco products (NTPs) among young adult smokers. METHODS: Focus groups with African Americans, non-Hispanic whites, and Hispanics. Risk ratings of light, regular, and menthol cigarettes and of NTPs and marijuana and cigarettes were compared. RESULTS: Participants tended to view light cigarettes as safer than regular cigarettes. Shisha and herbal products were rated as safer than traditional cigarettes, but there were differences in ratings by race/ethnicity, related to preferred cigarette variety. CONCLUSIONS: Health communication messages about the use of cigarettes and NTPs should consider risk perceptions about the products and racial/ethnic differences. PMID- 16712445 TI - Examining intentions to use CoQ10 amongst breast cancer patients. AB - OBJECTIVE: To determine factors that influence breast cancer patients' intentions to supplement with CoQ10. METHODS: A survey based upon the expanded rational expectations intentions model was completed by breast cancer outpatients (N=160). RESULTS: A significantly positive relationship existed between referent other (the influence specific people have in terms of an individual's behavior) and subjective norm (subject's perception of how people view a behavior). Beliefs, referent other, attitude, and subjective norm did have a significant effect on intention to use CoQ10. CONCLUSIONS: Health practitioners may address supplementation with breast cancer patients with a better understanding of what factors impact supplement use. PMID- 16712447 TI - Cyclooxygenase-2 inhibitors and cancer. PMID- 16712446 TI - Is smoking related to body image satisfaction, stress, and self-esteem in young adults? AB - OBJECTIVES: To examine the association of smoking and gender with body image satisfaction, perceived stress, and self-esteem in young adults. METHODS: Respondents completed a survey consisting of Perceived Stress Scale, Body-Areas Satisfaction Scale, Rosenberg Self-Esteem Scale, and the Positive and Negative Affect Schedule. Current smokers (n = 483) and never smokers (n = 973) are included. RESULTS: Smoking and female gender were independently associated with higher perceived stress (P < 0.001). Female gender was associated with lower body image satisfaction and lower self-esteem (P < 0.001). Current smoking was associated with lower self-esteem (P = 0.007). CONCLUSION: Smoking treatment should include stress management and self-esteem and body image improvement. PMID- 16712448 TI - Mechanisms leading to COX-2 expression and COX-2 induced tumorigenesis: topical therapeutic strategies targeting COX-2 expression and activity. AB - The biological role of COX-2, the inducible form of cyclooxygenase, is to convert arachidonic acid into prostaglandins (PGs) and thromboxanes (TXs). Overexpressed in many tumors, COX-2 plays a crucial role in cancer through synthesis of PGs which stimulate PGs receptors with subsequent enhancement of cellular proliferation, promotion of angiogenesis, inhibition of apoptosis, stimulation of invasion/motility, and suppression of immune responses. Depending on the tissue specificity and the cell type, several signaling pathways (Kinases, Rho, cGMP and Wnt), and transcription factors such as AP1, NFAT or NF-kappaB, are involved in COX-2 expression. In this review, we will describe mechanisms required by COX-2 metabolites to promote cancer development, and also the signaling pathways leading to COX-2 expression. In order to counteract the negative effects of COX-2 in cancerogenesis, chemicals interfering with COX-2 activity and expression were designed. We will give in the last part of this article, an overview of these potent chemicals interfering with the COX-2 signaling pathways involved in its expression or with its activity. PMID- 16712449 TI - The potential and rationale for COX-2 inhibitors in lung cancer. AB - Cyclooxygenase-2 (COX-2) overexpression is seen in many malignancies including lung cancer. Elevated tumor prostaglandin E2 (PGE2), a major COX-2 metabolite, levels have been implicated in angiogenesis, tumor growth and invasion, apoptosis resistance and suppression of anti-tumor immunity. Recent studies also revealed that PGE2 signaling may confer cells resistant to targeted growth factor receptor therapy by cross-activation of the receptor signaling pathway downstream components. Pre-clinical studies in animal tumor models have shown tumor reduction when animals are treated with COX-2 inhibitors and have demonstrated promising results when COX-2 inhibitors were combined with chemotherapeutic drugs. Based on these observations several ongoing clinical trials are currently evaluating COX-2 inhibitors as adjuvants with chemotherapy or radiation therapy in patients with advanced non-small cell lung cancer. Further understanding of the mechanisms of COX-2 in tumorigenesis and its interaction with other cellular pathways may highlight the new diagnostic, prognostic and therapeutic markers and facilitate future development of targeted strategies for lung cancer treatment and prevention. PMID- 16712450 TI - Interrelationships between cyclooxygenases and aromatase: unraveling the relevance of cyclooxygenase inhibitors in breast cancer. AB - Breast cancer is the most common cancer among women, and ranks second among cancer deaths in women. Approximately 60% of all breast cancer patients have hormone-dependent breast cancer, which contains estrogen receptors and requires estrogen for tumor growth. Estradiol is biosynthesized from androgens by the cytochrome P450 enzyme complex called aromatase. Aromatase is found in several tissues in the body and aromatase (CYP19) gene expression is regulated in a tissue-specific manner via use of alternative promoters. Aromatase transcript expression and activity in breast tumor tissue is greater than that in the normal breast tissue, and prostaglandins can increase CYP19 expression and aromatase activity in breast cancer cells. Cyclooxygenase (COX) is a key enzyme in the production of prostaglandins. Studies have shown higher levels of COX-2 isoform in breast cancer tissue when compared to normal breast tissue, and this is accompanied by high concentrations of prostaglandin E(2) (PGE(2)). Previous studies suggest a strong association between CYP19 gene expression and the expression of COX genes. While studies have shown that nonsteroidal anti inflammatory drugs (NSAIDs) have beneficial effects on breast cancer, the mechanism by which this occurs is still unclear. Studies have shown that COX inhibitors decrease aromatase activity in breast cancer cells and this effect starts at the transcriptional level. Real time PCR data shows that this molecular mechanism involves promoters I.4 and II, the promoters involved in the development of breast cancer. High levels of COX-2 expression result in higher levels of prostaglandin E(2) (PGE(2)), which in turn increases CYP19 expression through increases in intracellular cyclic AMP levels and activation of promoter II. Thus, PGE(2) produced via COX may act locally in paracrine and autocrine fashion to increase the biosynthesis of estrogen by aromatase in hormone dependent breast cancer development. PMID- 16712451 TI - The use of nimesulide and its analogues in cancer chemoprevention. AB - Non-steroidal anti-inflammatory drugs (NSAIDs), which are known to be cyclooxygenase (COX) inhibitors, have been reported to exert anti-proliferative and pro-apoptotic effects on a variety of cancer cells. Since the COX-2 isoform was found to be overexpressed in a many human cancers, a particular attention was paid on the possible use of selective COX-2 inhibitors in cancer chemoprevention. The present review focuses on the state of the art in cancer research developed with COX-2 preferential/selective inhibitors belonging to the family of N arylmethanesulfonamides, in particular nimesulide and NS-398. PMID- 16712452 TI - A computational view of COX-2 inhibition. AB - Selective inhibition of COX-2 provided a new class of anti-inflammatory, analgesic and antipyretic drugs with significantly reduced side effects and could also be an important strategy for preventing or treating a number of cancers. This review illustrates the molecular modeling methods used for the knowledge of the molecular mechanism of inhibition as well as for the design of selective compounds. PMID- 16712453 TI - Promotion of optimized protein therapy by bioconjugation as a polymeric DDS. AB - In recent years, clinical applications of recombinantly produced bioactive proteins such as cytokines have attracted attention. However, since these recombinant proteins are rather unstable in vivo, their clinical use as therapeutic agents requires frequent administration at a high dosage. This regimen disrupts homeostasis and results in severe side effects. To overcome these problems, bioactive proteins have been conjugated with water-soluble synthetic (WSS) polymeric carriers. Chemical modification of a protein with a WSS polymeric carrier (bioconjugation) regulates tissue distribution, resulting in a selective increase in its desirable therapeutic effects and a decrease in undesirable side effects. Among several drug delivery system (DDS) technologies, bioconjugation has been recognized as one of the most efficient methods for improving therapeutic potency of proteins. However, for further enhancement of the therapeutic potency and safety of conjugated bioactive proteins, more precise regulation of the in vivo behavior of each protein is necessary for selective expression of its therapeutic effect. Therefore, alternative WSS polymeric modifiers in which new functions such as targeting and controlled release of drugs can be added are required for further development of bioconjugated drugs. Recently, we have synthesized a novel polymeric drug carrier, poly(vinylpyrrolidone-co-dimethyl maleic anhydride) [PVD], which was a powerful candidate drug carrier for cancer therapy. In this review, we introduce useful information that enabled us to design polymeric drug carriers and their application for protein therapy. PMID- 16712454 TI - Biological effects of curcumin and its role in cancer chemoprevention and therapy. AB - Curcumin, a natural component of the rhizome of curcuma longa has emerged as one of the most powerful chemopreventive and anticancer agents. Its biological effects range from antioxidant, anti-inflammatory to inhibition of angiogenesis and is also shown to possess specific antitumoral activity. The molecular mechanism of its varied cellular effects has been studied in some details and it has been shown to have multiple targets and interacting macromolecules within the cell. Curcumin has been shown to possess anti-angiogenic properties and the angioinhibitory effects of curcumin manifest due to down regulation of proangiogenic genes such as VEGF and angiopoitin and a decrease in migration and invasion of endothelial cells. One of the important factors implicated in chemoresistance and induced chemosensitivity is NFkB and curcumin has been shown to down regulate NFkB and inhibit IKB kinase thereby suppressing proliferation and inducing apoptosis. Cell lines that are resistant to certain apoptotic inducers and radiation become susceptible to apoptosis when treated in conjunction with curcumin. Besides this it can also act as a chemopreventive agent in cancers of colon, stomach and skin by suppressing colonic aberrant crypt foci formation and DNA adduct formation. This review focuses on the various aspects of curcumin as a potential drug for cancer treatment and its implications in a variety of biological and cellular processes vis-a-vis its mechanism of action. PMID- 16712455 TI - Betulinic acid derivatives as anticancer agents: structure activity relationship. AB - Betulinic acid, a pentacyclic triterpene, is widely distributed throughout the tropics. It possesses several biological properties such as anticancer, anti inflammatory, antiviral, antiseptic, antimalarial, spermicidal, antimicrobial, antileshmanial, antihelmentic and antifeedent activities. However, betulinic acid was highly regarded for its anticancer and anti-HIV activities. Anticancer role of betulinic acid appeared by inducing apoptosis in cells irrespective of their p53 status. Due to high order safety in betulinic acid, a number of structural modifications carried out to improve its potency and efficacy. The C-1, C-2, C-3, C-4, C-20 and C-28 positions are the diversity centers in betulinic acid, and the derivatives resulted on various structural modifications at these positions screened for their anticancer activity. This review presents the structure activity relationship carried out on C-1, C-2, C-3, C-4, C-20, C-28, A-ring, D ring and E-ring modified betulinic acid derivatives. We have compiled the most active betulinic acid derivatives along with their activity profile in each series. Structure activity relationship studies revealed that C-28 carboxylic acid was essential for the cytotoxicity. The halo substituent at C-2 position in betulinic acid enhanced the cytotoxicity. Though the relation of the cytotoxicity with the nature of substituents at C-3 position could not be generalized but the ester functionality appeared to be a better substituent for enhancing the cytotoxicity. An interesting observation is that the three rings skeleton (A, B and C rings) had played an important role in eliciting anticancer activity, which could be a new molecular skeleton to design new anticancer drugs. PMID- 16712456 TI - Spectrum of radiopharmaceuticals in nuclear oncology. AB - A major field of interest in nuclear medicine is in vivo tumor characterization and measurement of biological processes at cellular and molecular levels by means of positron emission tomography (PET) or single photon emission computed tomography (SPECT). Functional imaging with radiopharmaceuticals represents a useful noninvasive tool to evaluate the biological status of the tumor and its progression. The properties of radiopharmaceuticals are exploited for initial staging of cancer, assessment of recurrent or residual disease and, more recently, considerable progress has been made in the field of the evaluation of tumor response to treatment. PET and SPECT can both detect changes in tumor activity caused by therapy or disease progression before any detectable change in tumor volume. Measurement of tumor response to therapy using PET and SPECT is the subject of intense investigations because it may result in individualization of treatment and may have a prognostic value for long-term outcome. This review focuses on the various methods used to monitor anticancer therapy with a variety of clinically approved or investigational tracers. We summarize the mechanisms of radiopharmaceutical uptake based on certain physiological activities affected by treatment: proliferation, apoptosis, hypoxia, angiogenesis and multidrug resistance (MDR). PMID- 16712457 TI - gamma-H2AX as a therapeutic target for improving the efficacy of radiation therapy. AB - Exposure to ionizing radiation (IR) results in the formation of DNA double strand breaks, resulting in the activation of phosphatidylinositol 3'-kinase-like kinases ATM, ATR and DNK-PKcs. A physiologically important downstream target is the minor histone H2A variant, H2AX, which is rapidly phosphorylated on Ser 139 of the carboxyl tail after IR. Recent work suggests that phosphorylated H2AX (gamma-H2AX) plays an important role in the recruitment and/or retention of DNA repair and checkpoint proteins such as BRCA1, MRE11/RAD50/NBS1 complex, MDC1 and 53BP1. H2AX-/- mouse embryonic fibroblasts are radiation sensitive and demonstrate deficits in repairing DNA damage compared to their wildtype counterparts. Cells treated with peptide inhibitors of gamma-H2AX demonstrate increased radiosensitivity following radiation compared with untreated irradiated cells. Analysis of the kinetics of gamma-H2AX clearance after IR or other DNA damaging agents reveals a correlation between increased gamma-H2AX persistence and unrepaired DNA damage and cell death. These data highlight the potential of post-translational modifications of chromatin as a therapeutic target for enhancing the efficacy of radiotherapy. Therapies that either block gamma-H2AX foci formation by inhibiting upstream kinase activity or that directly inhibit H2AX function may interfere with DNA damage repair processes and warrant further investigation as potential radiosensitizing agents. Agents that increase persistence of gamma-H2AX after IR are likely to increase unrepaired DNA damage. PMID- 16712458 TI - Molecular targets of ovarian carcinomas with acquired resistance to platinum/taxane chemotherapy. AB - Ovarian cancer of epithelial origin remains one of the most lethal malignancies despite response rates of more than 80% in first-line combination chemotherapy with platinum drugs and taxanes following surgery. Poor overall prognosis is mainly due to acquired resistance of the recurring tumor mass to initially used and other chemotherapeutic agents. Therefore, novel therapeutic approaches are based on concepts to prevent (improvement of tumor exposure to drugs) or circumvent drug resistance, e.g. with new drugs structurally related to the currently used cytotoxic agents, other types of cytotoxic substances, or with target-specific novel drugs interfering with signaling and apoptotic pathways. In addition, acquired molecular characteristics of drug resistant ovarian carcinoma cells can be defined by expression profiling at different stages of therapy and might be used as specific targets for tumor-suppressing drugs and prodrugs containing cytotoxic components. Revelation of mechanistic details of drug resistance also provides the basis for the development of therapies with novel or conventional antitumor drugs in combination with specific inhibitors able to re establish chemosensitivity. In this review, we summarize novel approaches in the treatment of ovarian cancer progressed to drug resistant stages and focus on the discussion of recently reported experimental and early clinical results with potentially useful strategies to overcome or modulate acquired drug resistance. PMID- 16712459 TI - Vitamin D receptor is a novel drug target for ovarian cancer treatment. AB - Ovarian cancer is the leading cause of death among gynecological malignancies in the US and the poor outcome of current treatments necessitates the development of novel therapeutic strategies to fight against it. Epidemiological data indicate a positive association between higher latitude and ovarian cancer incidence and mortality rates, suggesting that vitamin D insufficiency may contribute to ovarian cancer development. Recent studies in the authors' laboratory showed that multiple ovarian cancer cell lines respond to the active form of vitamin D, 1alpha,25-dihydroxyvitamin D(3), for growth suppression. Mechanistic studies identified vitamin D-regulated genes with established functions in ovarian tumorigenesis as mediators for the growth suppression. While increased p27 protein stability and transcriptional up-regulation of GADD45 are responsible for 1alpha,25-dihydroxyvitamin D(3)-induced cell cycle arrest at G1/S and G2/M checkpoints, respectively, the hormone-induced apoptosis in ovarian cancer cells involves the down regulation of the mRNA stability of telomerase catalytic subunit. More importantly, preclinical studies showed that the synthetic vitamin D analog EB1089 effectively suppressed the growth of human ovarian tumor xenografts in mice. The tumor suppression is associated with an increase in apoptotic rate and a decrease in cell proliferation, suggesting that the molecular information can be translated into ovarian tumor suppression in animals. Based on these studies, we conclude that the vitamin D receptor that mediates these anti-tumor effects represents a novel molecular target for the development of new drugs for ovarian cancer. We predict that receptor-based drug discovery will lead to the successful development of more potent and safer vitamin D analogs for the treatment of this deadly disease. PMID- 16712460 TI - Specialisation of the tropomyosin composition of actin filaments provides new potential targets for chemotherapy. AB - The actin microfilament network is important in maintaining cell shape and function in eukaryotic cells. It has a multitude of roles in cellular processes such as cell adhesion, motility, cellular signalling, intracellular trafficking and cytokinesis. Alterations in the organisation of the cytoskeleton and changes in cellular morphology, motility and adhesiveness are characteristic features of transformed cancer cells. For this reason cytoskeletal microfilaments have become promising targets for chemotherapy. In contrast to the microtubules, which have been targeted successfully with anti-tumour drugs such as Taxol-like compounds and the Vinca alkaloids, very few actin targeting drugs have been characterised. To date, no actin targeting drugs have been used in clinical trials due to their severe cytotoxicity. One reason for this cytotoxicity is that drugs such as the cytochalasins and latrunculins disrupt actin microfilaments in both non-tumour and tumour cells. To circumvent this problem, actin filament populations need to be targeted more specifically. Not all actin filaments are the same and there is growing evidence that within a cell there are different populations of actin filaments which are spatially organised into distinct cellular compartments each with a unique function. The structure and function of the actin cytoskeleton is primarily regulated by the associated actin binding proteins. Tropomyosin is an intrinsic component of most actin filaments and over 40 isoforms have been identified in non-muscle cells. Tm isoforms are spatially segregated and current evidence suggests that they can specify the functional capacity of the actin microfilaments. Therefore the composition of these functionally distinct actin filaments may be important in determining their stability and function within the cell. If actin filament populations can be discriminated and targeted based on their tropomyosin composition then this becomes a powerful approach for anticancer therapy. PMID- 16712461 TI - Gonadotropin-releasing hormone (GnRH) receptors in tumors: a new rationale for the therapeutical application of GnRH analogs in cancer patients? AB - Gonadotropin-Releasing Hormone (GnRH) is the hypothalamic decapeptide which plays a key role in the control of reproductive functions. By binding to specific receptors present on the pituitary gonadotropes, GnRH regulates gonadotropin release and, consequently, steroid hormone secretion from the gonads. When given continuously and at high doses, GnRH agonists suppress the pituitary gonadal axis through the down-regulation and desensitization of its own receptors. Based on this rationale, pituitary GnRH receptors represent the target for the successful utilization of GnRH agonists (that are more stable than the native peptide) for the treatment of hormone-dependent tumors (e.g., prostate, breast, endometrial, ovarian cancers). The observation that GnRH receptors are expressed in steroid dependent tumors, and that their activation reduces cell proliferation and metastatic behavior of cancer cell lines, both in vitro and in vivo (when inoculated into nude mice), indicates a possible additional and more direct antitumor activity for these compounds. Interestingly, GnRH receptors have been shown to be expressed also in androgen-independent prostate carcinoma, as well as in tumors that are not classically considered hormone-related (e.g., melanoma), suggesting a clinical utility of the administration of GnRH analogs also in these tumors. More recently, GnRH agonists have been proposed as useful carriers to target cytotoxic drugs or toxins to cancer cells displaying the specific GnRH receptors. A second form of GnRH (designated GnRH-II) has been discovered in most vertebrates, including humans. GnRH-II has been suggested to act through a 'putative' cognate type II GnRH receptor, which is distributed in different tissues, both normal and tumoral. In humans, a full-length functional type II GnRH receptor has not been found. Therefore, its functions as well as its possible utility as a molecular target for a GnRH-II based therapy in oncology still has to be clarified. This review will focus on the role of GnRH receptors in the control of tumor growth, progression and dissemination. It will also be discussed whether the presence of these receptors might represent an additional rationale for the clinical utility of GnRH analogs as anticancer drugs. PMID- 16712462 TI - Development and validation of an in silico P450 profiler based on pharmacophore models. AB - In today's drug discovery process, the very early consideration of ADME properties is aimed at a reduction of drug candidate drop out rate in later development stages. Apart from in vitro testing, in silico methods are evaluated as complementary screening tools for compounds with unfavorable ADME attributes. Especially members of the cytochrome P450 (P450) enzyme superfamily-- e.g. P450 1A2, P450 2C9, P450 2C19, P450 2D6, and P450 3A4-- contribute to xenobiotic metabolism, and compound interaction with one of these enzymes is therefore critically evaluated. Pharmacophore models are widely used to identify common features amongst ligands for any target. In this study, both structure-based and ligand-based models for prominent drug-metabolizing members of the P450 family were generated employing the software packages LigandScout and Catalyst. Essential chemical ligand features for substrate and inhibitor activity for all five P450 enzymes investigated were determined and analyzed. Finally, a collection of 11 pharmacophores for substrates and inhibitors was evaluated as an in silico P450 profiling tool that could be used for early ADME estimation of new chemical entities. PMID- 16712463 TI - Rational design approaches to chemical libraries for hit identification. AB - Sequencing of the human genome along with developments in combinatorial synthesis and high-throughput biological screening provide unparallel opportunities to drug discovery. It has been noted that the increased number of synthesized and annotated compounds did not yield the expected increase in number of viable drug candidates. To address this problem, several novel computation technologies have emerged for making combinatorial library design cost-effective. Of particular interest for the modern drug discovery are the structure-based or target-based methods that use structural information about target proteins and their small molecule ligands. In this work, we provide an overview of selected advances in computational algorithms for the rational selection of molecule libraries for the synthesis, with emphasis on structure-based approaches. These include a fusion of scaffold-linking method and combinatorial library design, pharmacophore matching and informative library design, and search by 3-D tree topological descriptors. PMID- 16712464 TI - Anti-inflammatory and anti-oxidant activity of a new class of phenyl-pyrazolone derivatives. AB - The anti-inflammatory activity of a new class of phenyl-pyrazolone derivatives, structurally related to phenidone, has been evaluated using the Croton oil ear test in mice as model of acute inflammation. Derivative 5h reduces the percentage of oedema similarly to indomethacin and more efficiently than phenylbutazone. The anti-inflammatory activity of these two reference drugs depends on their COX inhibition, but for the synthesized derivatives it has not been demonstrated a significant COX or LOX inhibition, as previously reported. While the anti inflammatory activity of phenidone is correlated to its anti-oxidant properties, the redox potential of these compounds appears not decisive in the inflammatory process inhibition. In order to investigate the mechanism of action for these compounds, we quantified their anti-oxidant activity and the lipophilicity, and a relationship between the calculated logP and the percentage of oedema reduction was found. We hypothesize that the anti-inflammatory activity, recorded in vivo, could be related to lipophilic parameter of these compounds. PMID- 16712465 TI - Fine-grained statistical torsion angle potentials are effective in discriminating native protein structures. AB - Modelling of drug targets requires the reliable selection of an accurate and representative structure from large ensembles of alternate models. Statistical potentials developed to discriminate native protein structures generally represent pairwise interactions between atoms, which are less sensitive to local conformational details. The discrimination of local distortions is therefore particularly difficult. Local interaction preferences, expressed through torsion angles, are rarely used, as some controversy exists in the literature regarding their discrimination power. The present study aims to benchmark the efficiency of different implementations of torsion angle propensities for selecting the native structure from ensembles of well-constructed decoys. Several statistical potentials derived from fine-grained discretisations of torsion angle space are constructed and evaluated. Results from a comparison with nine widely used statistical scoring functions show the torsion angle potentials to be more effective in recognising native structures and to improve with the number of torsion angles considered. These data suggest local structural propensities to be important for estimating the overall quality of native-like models. PMID- 16712466 TI - Advanced glycation end products (AGEs) and their receptor (RAGE) system in diabetic retinopathy. AB - Vascular complications are a leading cause of blindness, end-stage renal failure, a variety of neuropathies and accelerated atherosclerosis, which could account for disabilities and high mortality rates in patients with diabetes. There is a growing body of evidence that formation and accumulation of advanced glycation end products (AGEs) progress during normal aging, and at an extremely accelerated rate in diabetes, thus being involved in the pathogenesis of diabetic vascular complications. Furthermore, the interaction by AGEs of their receptor, RAGE, activates down-stream signaling and evokes inflammatory responses in vascular wall cells. Therefore, inhibition of AGE formation or blockade of the RAGE signaling may be a promising target for therapeutic intervention to prevent diabetic vascular complications. This review discusses the molecular mechanisms of diabetic retinopathy, especially focusing on the AGE-RAGE system. Several types of inhibitors of the AGE-RAGE system and their therapeutic implications are also reviewed here. PMID- 16712468 TI - Human umbilical cord blood biology, transplantation and plasticity. AB - As the significance of hematopoietic stem cell transplantation (HSCT) is constantly rising, the scarcity of matched donors is proving to be a troubling issue. Cord blood (CB) is an important source of stem cells (SC) for transplantation. It has been used in the last two decades for approximately 4500 transplantations. Its collection, cryopreservation, banking and thawing techniques pose unique challenges to clinicians and researchers CB has abundant stem cell with impressive proliferative capacity. On the other hand, CB's immunological system has a naive and more tolerant nature. Except for the biological aspects, few ethical issues have become a concern for transplantation teams who use CB. There are few advantages of CB over bone marrow, especially the lower rates of acute and chronic graft-versus-host disease (GVHD) after transplantation. On the other hand, there are relatively high rates of early treatment related mortality in cord blood transplantation (CBT). This is related to the small nucleated cell (NC) dose infused from each CB unit. The clinical experience in CBT, especially in children, is encouraging. When using adequate number of NC/kg, results in CBT for malignant and non-malignant diseases are comparable to bone marrow transplantation (BMT). In this article, a comprehensive review of the largest scale studies is presented. There is a continuous search for an optimal way to deal with delayed engraftment of CB and its implication. The current investigational, and also first clinical trials using diverse methods to overcome high rates of TRM are reviewed. Almost twenty years after the first CBT was preformed, many advocate for a routine parallel search, BM and CB, for unrelated donor. Future uses of CB might also be in the field of gene transfer and non hematopoietic injured tissues repair. PMID- 16712467 TI - (Q)SAR studies to design new human choline kinase inhibitors as antiproliferative drugs. AB - Most of the signal transduction pathways are mediated by protein kinases regulating every aspect of cell function. Mutations which deregulate their expression or their function or both result in cancers. Therefore, protein kinase inhibitors have become the focus of development of new therapies for cancer. A comprehensive review of Choline kinase (ChoK) was published by us in 2003. Since then, molecular information of ChoK inhibitors has been accumulated. In this review, we intend to summarize the new lines of evidence that will include the design of the most active antiproliferative agents so far described against ChoK. Studies have been aimed at the establishment of structure-activity relationships and the structural parameters that define ChoK inhibitory and antiproliferative activities of a set of twenty-five acyclic biscationic pyridophane and forty acyclic biscationic quinolinephane compounds. The corresponding QSAR equation was obtained for the whole set of bisquinolinium compounds for the antiproliferative activity, taking into consideration the electronic parameter sigma(R) of R(4), the molar refractivity (MR) of R(8), and the lipophilic parameters clog P and pi(linker). The most potent antiproliferative agent shows an IC(50) = 0.45 microM, predicted by the QSAR equation, whilst its experimental value is IC(50) = 0.20 microM. Finally, toxicity assays were performed for the most promising compounds because of their interesting antiproliferative activities [IC(50 HT-29) = 0.70, 0.80, 1.50 and 1.90 microM] and low toxicity [LD(50) = 16.7, 12.5, > 25 and > 20 mg/kg of mouse]. These biological activities justify further analysis for antitumoral assays under in vivo conditions. PMID- 16712469 TI - Recent advances in small molecule antagonists of the corticotropin-releasing factor type-1 receptor-focus on pharmacology and pharmacokinetics. AB - Corticotropin-releasing factor (CRF), a 41-amino acid peptide, has been recognized as an important factor mediating stress. Efforts to discover small molecule antagonists of the CRF type-1 receptor (CRF(1)-R) for potentially novel treatment of anxiety and depression started in the early 1990's. Although highly potent in vitro and efficacious in animal models, early reported compounds such as CP-154,526 and NBI-27914 are highly lipophilic and possess high plasma protein and tissue binding, long elimination half life, and toxicity, likely due in part to accumulation in tissues. Recently, several laboratories have reported potent CRF(1)-R antagonists with improved physicochemical properties. Compounds such as DMP696, NBI-30775/R121919 and R278995/CRA0450 possess at least one additional polar group in their structures and are therefore less lipophilic than the earlier compounds, while still maintaining high potency. For example, DMP696 has a K(i) value of 1.7 nM and a cLogP of 3.2, which is similar to CP-154,526 in potency but about 4-log units lower in partition coefficient. Despite its high plasma protein binding (98.5% in rat), DMP696 occupies over 50% of brain CRF(1)-R at a total plasma concentration above 100 nM, which is consistent with the doses that produce anxiolytic effects in the rat defense withdrawal test of anxiety. This article will review small molecule CRF(1)-R antagonists by focusing on their pharmacological and pharmacokinetic properties. In addition, the pharmacology of small molecules binding to the CRF(1) receptor will be discussed. An orally available compound with desirable properties in these categories will have a good chance to be developed into a novel treatment for anxiety and depression which may be devoid of the side effects of existing antidepressant treatments. PMID- 16712470 TI - Receptor-ligand binding sites and virtual screening. AB - Within the pharmaceutical industry, the ultimate source of continuing profitability is the unremitting process of drug discovery. To be profitable, drugs must be marketable: legally novel, safe and relatively free of side effects, efficacious, and ideally inexpensive to produce. While drug discovery was once typified by a haphazard and empirical process, it is now increasingly driven by both knowledge of the receptor-mediated basis of disease and how drug molecules interact with receptors and the wider physiome. Medicinal chemistry postulates that to understand a congeneric ligand series, or set thereof, is to understand the nature and requirements of a ligand binding site. Likewise, structural molecular biology posits that to understand a binding site is to understand the nature of ligands bound therein. Reality sits somewhere between these extremes, yet subsumes them both. Complementary to rules of ligand design, arising through decades of medicinal chemistry, structural biology and computational chemistry are able to elucidate the nature of binding site-ligand interactions, facilitating, at both pragmatic and conceptual levels, the drug discovery process. PMID- 16712471 TI - Inhibitors of HIV-1 Tat-mediated transactivation. AB - The transactivation responsive (TAR) RNA is the 5'-leader sequence of the HIV-1 mRNA genome and interacts with the Tat protein during transcription. Tat and the positive transcription elongation factor (P-TEFb) complex bind to TAR to promote efficient transcription of the full-length HIV genome. In the absence of the TAR.Tat.P-TEFb interaction, viral transcription is inefficient, which makes this RNA-protein complex an important target for therapeutic intervention of HIV replication. Inhibitors of HIV-1 transactivation mainly target: 1) TAR RNA, 2) Tat protein and 3) Tat.P-TEFb complex. 1) Compounds against TAR RNA are the most numerous: besides cationic peptides, which were initially developed, recent advances in TAR binding inhibitors include oligonucleotide based-agents and small molecules. Specific research efforts are currently underway to increase cellular uptake. 2) By targeting the Tat protein, both transactivation and other Tat mediated intra/extracellular functions are affected. Various biopolymeric drugs are reported to effectively inhibit Tat activity. In addition, Tat-targeted antibodies have recently been developed. 3) Intracellular proteins have been discovered to disrupt Tat.P-TEFb interaction, raising the chance of inhibiting HIV-1 transcription via novel mechanisms. PMID- 16712472 TI - Nuclear translocation during the cross-talk between cellular stress, cell cycle and anticancer agents. AB - The function of many endogenous molecules in all eukaryotic cells depends on their subcellular localisation, being active when localized in one cellular compartment and inactive in another. Translocation or re-localization of mislocalized components in the optimal subcellular site may contribute to the development of novel cancer therapies and to the re-evaluation of conventional treatment. For instance, various agents are able to entrap cytoplasmic anti apoptotic pathways to the nucleus, thus activating apoptosis. Moreover, amongst the factors identified so far, the optimal location of the tumor suppressor p53 for promoting cell arrest and apoptosis seems to be the nucleus, while the nuclear factor kappa B (NFkappaB) is desirable to stay in the cytoplasm. Thus, the mechanisms of nuclear translocation of endogenous signaling components, like p53, NFkappaB and various heat shock proteins (HSPs), may serve as targets for pharmacological intervention, without excluding the possible role of uptake and active transport into the nucleus of extracellular proteins. PMID- 16712473 TI - Current advances in antifungal targets and drug development. AB - Fungi are one of the most neglected pathogens apparent from the fact that the Amphotericin B, a polyene antibiotic, discovered way back in 1956 is still used as a "gold standard" for antifungal therapy. Past two decades have witnessed a dramatic rise in the incidences of life threatening systemic fungal infections. This can be ascribed to the increase in the number of immuno-compromised patients due to rise in HIV infected population, cancer chemotherapy and indiscriminate use of antibiotics. Majority of clinically used antifungals suffer from various drawbacks in terms of toxicity, efficacy and cost, and their frequent use has led to the emergence of resistant strains. Hence, there is a great demand for novel antifungals belonging to wide range of structural classes, selectively acting on novel targets with fewer side effects. This article aims at reviewing recent efforts made towards discovering novel antifungal drug targets and investigational molecules acting on them. PMID- 16712474 TI - Platinum group antitumor chemistry: design and development of new anticancer drugs complementary to cisplatin. AB - In the next two decades, the world is expected to see around 20 million cases of cancer. Moreover, the types of cancer will vary considerably from country to other. Therefore, all efforts will be needed to face such a vast diversity of problems. With current annual sales of about $500 millions, the platinum(II) complex known as cisplatin [cis-(NH3)2PtCl2] is still one of the most effective drugs to treat testicular, ovarian, bladder and neck cancers. Since it was launched in 1978 there has been a rapid expansion in research to find new, more effective metal-based anticancer drugs and to study their interactions with biological systems. This study gives an up to date overview of the anticancer chemistry of the platinum group elements platinum, palladium, and nickel with an emphasis on the new strategies used in the development of new antitumor agents. Methodologies for application of bulky aromatic or aliphatic nitrogen ligands, chiral organic moieties, chelates containing other donor atoms than nitrogen, and biologically active ligands in the design of agents analogous to cisplatin are presented. The review also aims to highlight the class of the unconventional complexes that violate the empirical structure-activity rules (SAR) of platinum compounds and the common features and structural differences between the most successful anticancer complexes that are currently in human clinical trials. PMID- 16712475 TI - From gut homeostasis to cancer. AB - The mammalian intestine has one of the highest turnover rates in the body. The intestinal epithelium is completely renewed in less than a week. It is divided into spatially distinct compartments in the form of finger-like projections and invaginations that are dedicated to specific functions. Intestinal cells are constantly produced from a stem cell reservoir that gives rise to proliferating transient amplifying cells, which subsequently differentiate and migrate to the correct compartment before dying after having fulfilled their physiological function. In recent years, a substantial body of evidence has accumulated to support the concept that signaling pathways known to be crucial for embryonic development of multiple organisms play a critical role in tightly regulating and controlling the self-renewing process of the intestine. Moreover, the same pathways appear to be deregulated in several hereditary and sporadic colorectal cancer syndromes due to activating and/or inactivating mutations of key components of such pathways. In this review we discuss recent findings demonstrating that differentiation and homeostasis of the intestine are controlled by developmental pathways such as Wnt, Notch, TGF-beta and Hedgehog, and illustrate how their deregulation contributes to intestinal neoplasia. PMID- 16712476 TI - Lymphoma and the control of B cell growth and differentiation. AB - It is now widely accepted that lymphomagenesis is a multistep transformation process. A number of genetic changes and environmental and infectious factors contributing to the development and malignant progression of B-cell lymphoproliferative disorders are well documented. Reciprocal chromosomal translocations involving the immunoglobulin loci are a hallmark of most mature B cell lymphomas and lead to dysregulated expression of proto-oncogenes (c-myc) important for cell proliferation or genes involved in cell cycle progression (cyclin D1), differentiation block (bcl-6, PAX5) and cell survival (bcl-2, NF kappaB). In addition, genetic alterations that inactivate tumor suppressor genes (p53, p16) have been frequently detected in some lymphoma tissues. Many of these genes are normally regulated by signals from the B cell antigen receptor. The high prevalence of bacterial and viral infection in lymphoma patients supports the hypothesis that infectious agents may play a contributory role in the development and evolution of B cell lymphoproliferative disorders by either directly inducing polyclonal B cell hyperactivation (EBV, HCV), or providing a chronic antigenic stimulus (EBV, HCV, HBV, H. pylori), or mimicking B cell antigen receptor signaling (EBV, HCV, HHV8), although whether these are causative factors or they are secondary to genetic changes in lymphomagenesis remains to be defined. Stimulatory signals from reactive T cells, local cytokines and growth factors can also contribute, to some extent, to the progression of transformation. Modulation of B cell antigen receptor signaling therefore emerges as a potentially powerful strategy for controlling the growth of certain B cell lymphomas. PMID- 16712477 TI - Malaria: therapy, genes and vaccines. AB - Malaria kills over 3,000 children each day. Modern molecular and biochemical approaches are being used to help understand and control Plasmodium falciparum, the parasite that causes this deadly disease. New drugs are being invented for both chemoprophylaxis and therapeutic treatments and their use is discussed along side that of the more commonly used treatments. Classical genetic crosses coupled with molecular analysis of gene loci are use to explain the genetics behind the development of specific drug resistances that the parasites have naturally developed. Rapid advances in DNA sequencing techniques have allowed the compete sequencing of the P. falciparum and several other rodent malaria parasite genomes. Proteomics and computational analysis of these vast databanks are being used to model and investigate the three-dimensional structure of many key malaria proteins in an attempt to facilitate drug design. Recombinant protein expression in bacteria and yeast coupled with cGMP purification technologies and conditions have lead to the recent availability of several dozen malaria protein antigens for human-use Phase I and Phase II vaccine trials. Drug companies, private foundations, and key government agencies have contributed to the coordinated efforts needed to test these antigens, adjuvants and delivery methods in an effort to find an effective malaria vaccine that will prevent infection and disease. PMID- 16712479 TI - Small RNAs and non-small cell lung cancer. AB - Patients with non-small cell lung cancer (NSCLC) are commonly diagnosed with advanced disease and have limited therapeutic options. Experimental treatment approaches including small molecule targeted therapeutics, gene modified tumor vaccines, and viral-based gene therapy have induced tumor regression in a small proportion of patients, suggesting that advanced NSCLC is susceptible to molecular perturbations. RNA interference (RNAi) has generated considerable excitement as a potential cancer therapeutic application. RNAi is the process by which small, double stranded RNA molecules (small interfering RNA, or siRNA) can initiate sequence-specific, post-transcriptional gene silencing (PTGS). Cancer growth inhibition was attained through siRNA-knockdown of unique or overexpressed cancer oncogenetic messages that are relevant to NSCLC pathophysiology. As with other loss-of-function cancer gene therapy approaches, clinical efficacy of siRNA depends largely on the extent of cell target coverage at the locoregional and/or systemic level. Cationic liposomes as well as viral vectors have been used successfully for siRNA delivery. However, viral delivery may have more immediate relevance due to its wider clinical acceptance in the cancer gene therapy arena. We advocate the use of conditional replicative, oncolytic adenovirus for siRNA delivery, which offers potential benefits of restricted and renewable siRNA expression within the tumor microenvironment, and an additive anti-tumor outcome through viral oncolysis and siRNA-mediated oncogene-silencing, which we have demonstrated with the A549 NSCLC cell line. Several oncolytic adenoviral constructs are potentially applicable clinical platforms with proven infectivity and safety, which are feasible also for the delivery of microRNAs (miRNA), a recently discovered group of endogenous, small RNA with PTGS activity that is downregulated in lung cancer. PMID- 16712480 TI - TAGE (toxic AGEs) theory in diabetic complications. AB - Diabetic complication is a leading cause of acquired blindness, end-stage renal failure, a variety of neuropathies and accelerated atherosclerosis. Chronic hyperglycemia is initially involved in the pathogenesis of diabetic micro- and macro-vascular complications via various metabolic derangements. High glucose increased production of various types of advanced glycation end-products (AGEs). Recently, we found that glyceraldehyde-derived AGEs (AGE-2) play an important role in the pathogenesis of angiopathy in diabetic patients. There is considerable interest in receptor for AGEs (RAGE) found on many cell types, particularly those affected in diabetes. Recent studies suggest that interaction of AGE-2 (predominantly structure of toxic AGEs; TAGE) with RAGE alters intracellular signaling, gene expression, release of pro-inflamatory molecules and production of reactive oxygen species (ROS) that contribute towards the pathology of diabetic complications. We propose three pathways for the in vivo formation of AGE-2 precursor, glyceraldehyde, such as i) glycolytic pathway, ii) polyol pathway, and iii) fructose metabolic pathway. Glyceraldehyde can be transported or can leak passively across the plasma membrane. It can react non enzymatically with proteins to lead to accelerated formation of TAGE at both intracellularly and extracellularly. In this review, we discuss the molecular mechanisms of diabetic complications, especially focusing on toxic AGEs (TAGE) and their receptor (RAGE) system. PMID- 16712482 TI - Mechanisms of ERK1/2 regulation by seven-transmembrane-domain receptors. AB - Control of cell growth and differentiation has long been a focus of intense research interest, particularly in the context of cancer therapeutics. The evolutionarily-conserved extracellular signal-regulated kinases 1 and 2 (ERK1/2) are serine-threonine kinases that respond to a wide range of mitogens and growth factors to initiate changes in cellular proliferation and differentiation, and are the most important members of the mitogen-activated protein kinase (MAPK) family in terms of seven transmembrane-domain receptor (7TMR)-mediated regulation of mitogenic processes. Regulation of the ERK1/2 signaling cascade by 7TMRs is highly complex and cell type-specific. Recent advances in our knowledge of this effector pathway have revealed that its regulation is at least partly independent of traditional G protein-mediated actions arising from the stimulation of 7TMRs. This review summarizes the current position of our knowledge of ERK1/2 regulation, and illustrates the wealth of potential targets available for the development of new strategies for the treatment of proliferative and other ERK related disorders. PMID- 16712478 TI - Toll-like receptors and corneal innate immunity. AB - The ocular surface is constantly exposed to a wide array of microorganisms. The ability of the cornea to recognize pathogens as foreign and eliminate them is critical to retain its transparency, hence preservation of sight. In the eye, as in other parts of the body, the early response against invading pathogens is provided by innate immunity. Corneal innate immune system uses a series of pattern recognition receptors to detect the presence of pathogens thus allowing for rapid host defense responses to invading microbes. A key component of such receptors is the "Toll-like receptors" (TLRs), which have come to occupy the center stage in innate immunity against invading pathogens. An increasing number of studies have shown that TLRs are expressed by a variety of tissues and cells of the eye and play an important role in ocular defense against microbial infection. Here in this review we summarize the current knowledge about TLR expression in human eye with main emphasis on the cornea, and discuss the future directions of the field. PMID- 16712483 TI - G proteins in drug screening: from analysis of receptor-G protein specificity to manipulation of GPCR-mediated signalling pathways. AB - Seven transmembrane G protein coupled receptors (7TM GPCRs) represent one of the largest gene familes in the human genome. Because of the size of the GPCR family, their proven history of being valuable targets for small molecule drug design, the fact that the absolute number of GPCRs that are targets for current medicines represents only a small fraction of the total encoded by the human genome, and that ligands for GPCRs do not have to enter the cell to exert their function, it is very likely that GPCRs will remain major targets for the pharmaceutical industry in the foreseeable future. Despite recent evidence indicating that GPCRs can provide information to cells, that does not require activation of G proteins ("signaling at zero G"), most of the GPCRs known to date function via interaction with and activation of heterotrimeric (alphabetagamma) G proteins. Thus, assay systems translating ligand modulation of GPCRs into G protein-dependent intracellular responses are a key component of both basic research and the drug discovery process. This article will review the current knowledge and recent progress in understanding molecular aspects of specific receptor-G protein recognition. It will also highlight how the knowledge generated by such studies can be transformed into assay systems for GPCR drug discovery. PMID- 16712484 TI - Integrative functional assays, chemical genomics and high throughput screening: harnessing signal transduction pathways to a common HTS readout. AB - Chemical genomics is a drug discovery strategy that relies heavily on high throughput screening (HTS) and therefore benefits from functional assay platforms that allow HTS against all relevant genomic targets. Receptor Selection and Amplification Technology (R-SAT) is a cell-based, high-throughput functional assay where the receptor stimulus is translated into a measurable cellular response through an extensive signaling cascade occurring over several days. The large biological and chronological separation of stimulus from response provides numerous opportunities for enabling assays and increasing assay sensitivity. Here we review strategies for building homogeneous assay platforms across large gene families by redirecting and/or amplifying signal transduction pathways. PMID- 16712485 TI - Glucagon and glucagon-like peptide receptors as drug targets. AB - Glucagon and the glucagon-like peptides are derived from a common proglucagon precursor, and regulate energy homeostasis through interaction with a family of distinct G protein coupled receptors. Three proglucagon-derived peptides, glucagon, GLP-1, and GLP-2, play important roles in energy intake, absorption, and disposal, as elucidated through studies utilizing peptide antagonists and receptor knockout mice. The essential role of glucagon in the control of hepatic glucose production, taken together with data from studies employing glucagon antagonists, glucagon receptor antisense oligonucleotides, and glucagon receptor knockout mice, suggest that reducing glucagon action may be a useful strategy for the treatment of type 2 diabetes. GLP-1 secreted from gut endocrine cells controls glucose homeostasis through glucose-dependent enhancement of beta-cell function and reduction of glucagon secretion and gastric emptying. GLP-1 administration is also associated with reduction of food intake, prevention of weight gain, and expansion of beta-cell mass through stimulation of beta-cell proliferation, and prevention of apoptosis. GLP-1R agonists, as well as enzyme inhibitors that prevent GLP-1 degradation, are in late stage clinical trials for the treatment of type 2 diabetes. Exenatide (Exendin-4) has been approved for the treatment of type 2 diabetes in the United States in April 2005. GLP-2 promotes energy absorption, inhibits gastric acid secretion and gut motility, and preserves mucosal epithelial integrity through enhancement of crypt cell proliferation and reduction of epithelial apoptosis. A GLP-2R agonist is being evaluated in clinical trials for the treatment of inflammatory bowel disease and short bowel syndrome. Taken together, the separate receptors for glucagon, GLP-1, and GLP-2 represent important targets for developing novel therapeutic agents for the treatment of disorders of energy homeostasis. PMID- 16712486 TI - Cannabinoid receptors as therapeutic targets. AB - The cannabinoid receptors CB1 and CB2 are family A, G-protein Coupled Receptors that mediate the effects of cannabinoids, a class of compounds that are so named because the first members were isolates of the cannabis plant. In recent history, there has been much anecdotal evidence that the potent and diverse physiological responses produced by these compounds can be turned to therapeutic benefit for a wide variety of maladies. The remarkable abundance of cannabinoid receptors and the discovery of several endogenous ligands along with enzyme and transporter proteins for which they are substrates, suggests that an endogenous cannabinoid neuromodulatory system is an important mediator of biological function. For these reasons CB1 and CB2 receptors are attractive targets for the design of therapeutic ligands. The action of these receptors, however, may also be modulated by manipulating the enzymes and membrane transporters that regulate the endogenous ligands. Despite the range of physiological processes and activities that are mediated by cannabinoid receptors, it is clear that it is possible to produce ligands that result in differential responses. In this paper, we review the pharmacophoric elements that lead to these differential responses and in order to discuss them in context we present an overview of structural aspects governing cannabinoid receptor function, the cannabinergic system and its physiological functions. PMID- 16712487 TI - A role for information collection, management, and integration in structure function studies of G-protein coupled receptors. AB - Elucidation of protein function is greatly facilitated by the availability of an atomic resolution structure or a reliable molecular model. The difficulty of obtaining atomic resolution structures of membrane proteins in general, and of G protein coupled receptors (GPCRs) in particular, has made the information available from sequence analysis, mutagenesis, and the literature on related GPCRs exceptionally important. Here, we review previous studies of GPCR structure function from the perspectives of sequence analysis, management of mutagenesis and ligand binding data, and literature data mining. The knowledge derived from these information resources not only constitutes the prerequisites for reliable molecular modeling, but also can provide other insights into GPCR functions. Finally, we review approaches for information integration and applying knowledge discovery techniques to structure-function studies of GPCRs, including molecular modeling itself. PMID- 16712488 TI - Screening the receptorome yields validated molecular targets for drug discovery. AB - With the recently completed sequencing and annotation of the human genome, it has become clear that a significant portion of the genome encodes signal-transducing molecules including receptors, protein kinases, ion channels, transporters and coupling proteins. This review focuses on membrane-localized receptors, which represent the largest single group of signal-transducing molecules. Indeed, one can estimate that nearly 10% of the human genome encodes membrane-localized receptors (e.g. G-protein coupled receptors, ligand-gated ion channels and transporters). We have defined that portion of the human genome that encodes 'receptors' the receptorome. In this article, we will demonstrate how the massively parallel screening of the receptorome provides a facile and under utilized screening platform for drug discovery. Using case studies, we will show how receptorome-based screening elucidates the mechanisms responsible for serious side-effects of both approved and investigational medications. Additionally, we will provide evidence that receptorome-based screening provides insights into novel therapeutic indications of approved medications and serves to validate targets for therapeutic drug discovery. PMID- 16712489 TI - Co-ordinated covalent modification of G-protein coupled receptors. AB - The G-protein coupled receptor (GPCR) gene family represents one of the largest families in the mammalian genome. The flexibility of signalling and widespread tissue distribution of these receptors has allowed GPCRs to be employed in the physiological regulation of nearly all biological functions. This, coupled with the fact that it is possible to chemically produce highly specific ligands to these receptors have made GPCRs attractive targets for pharmacological intervention in a wide variety of disease states. When targeting GPCRs in therapeutic drug design it is traditional, and eminently sensible, to focus on ligands that will provide agonism, antagonism or allosteric modulation. However, as more is understood of the mechanisms that regulate GPCRs, and in particular the dynamic covalent modifications that might endow tissue specific functions, then these regulatory processes may provide alternative targets for GPCR drug discovery. In this review we consider three of the covalent modifications which are considered to regulate the function of GPCRs namely; receptor phosphorylation, palmitoylation and ubiquitination. In particular, we will describe the mechanisms of modification, the functional consequences and the relationship between these three covalent modification events. PMID- 16712491 TI - Approaches to palliative therapies for Alzheimer's disease. AB - Alzheimer's disease is a progressive neurodegenerative disease characterized by gradual and increasing loss of cognitive function and behavioral abnormalities. The formation of beta-amyloid plaques and neurofibrillary tangles are recognized as the key pathologies of the disease. Changes in the levels of various key neurotransmitters has been noted in patients with Alzheimer's disease and may represent the earliest biochemical casualty, preceding or signifying the onset of the disease. Over the last 20 years a number of approaches to the palliative treatment of Alzheimer's disease have been scrutinized. The majority of effort has been focused on cognitive dysfunction, as this is the initial and key debilitating symptom of the disease. The identification and commercial development of the acetylcholinesterase inhibitors has, until recently, virtually dominated the field, and although efficacy has been demonstrated, the clinical results suggest alternate approaches are warranted. This review will highlight those palliative approaches that have focused on the improvement of learning and memory and not on the disease-modifying strategies of the beta-amyloid, tau phosphorylation or other neurodegenerative hypotheses. PMID- 16712492 TI - Human beta-secretase (BACE) and BACE inhibitors: progress report. AB - A key step in the processing of the integral membrane protein APP, or Amyloid Precursor Protein is through the proteolytic cleavage by the enzyme beta Secretase (BACE). The proteolysis of APP by BACE, followed by subsequent C terminal cleavage(s) by gamma-secretase, results in the formation of the amyloid beta (Abeta) peptide. The principal component of the neuritic plaque found in the brains of Alzheimer's Disease (AD) patients is Abeta which is a neurotoxic and highly aggregatory peptide segment of APP. The amyloid hypothesis holds that the neuronal dysfunction and clinical manifestation of AD is a consequence of the long term deposition and accumulation of 40-42 amino-acid long Abeta peptides, and that this process leads to the onset and progression of AD. Due to the apparent causal relationship between Abeta and AD, the so-called "secretases" that produce Abeta have been targeted for development of inhibitors that might serve as therapeutic agents for treatment of this dreaded, and ever more prevalent disease. Herein will be discussed our current understanding of BACE, its role in the formation of neuritic plaques and the known inhibitors of the enzyme. PMID- 16712493 TI - Tau therapeutic strategies for the treatment of Alzheimer's disease. AB - The two classical pathological hallmarks of Alzheimer's disease are deposits of aggregated beta-amyloid (Abeta) peptide and neurofibrillary tangles composed of hyperphosphorylated tau protein. In addition to Abeta pathology, an invariant trait of Alzheimer's disease, disruption of tau processing is a necessary event in the neurotoxic cascade which eventually leads to neuronal death and subsequent dementia. Tau is a neuronal, microtubule-bound protein which becomes hyperphosphorylated as a result of an imbalance of the kinase and phosphatase activities which normally tightly regulate its phosphorylation. In addition to this pathogenic hyperphosphorylation, tau dissociates from microtubules and self aggregates to form insoluble oligomers which progress to the macroscopic tangles evident in post mortem Alzheimer's disease tissue. Subsequent toxicity may ensue either as a direct toxic effect of free tau oligomers or as a result of altered microtubule-dependent processes. In order to intervene pharmacologically in this disease process, much effort has been expended in order to identify and inhibit the kinases responsible for pathogenic hyperphosphorylation and many candidate kinases have been investigated including glycogen synthase kinase (GSK-3), cyclin dependant kinase-5 (Cdk-5), MAPK family members (extracellular signal-regulated kinases 1 and 2 [Erk-1 and 2], MEK [MAP kinase kinase], c-Jun NH(2)-terminal kinases (JNKs) and p38), casein kinase, calcium calmodulin-dependant kinase II (CaMK-II), microtubule affinity regulating kinase (MARK), protein kinase A (PKA/cAMP-dependant protein kinase) and others. Focus has also fallen upon the role of the phosphatases responsible for dephosphorylation of tau. This review will describe the tau-related etiology of Alzheimer's disease and other tauopathies as well as the therapeutic strategies to inhibit the hyperphosphorylation of tau. PMID- 16712494 TI - The role of amyloid-beta derived diffusible ligands (ADDLs) in Alzheimer's disease. AB - The amyloid-beta (Abeta) cascade hypothesis of Alzheimer's disease (AD) has dominated research and subsequent therapeutic drug development for over two decades. Central to this hypothesis is the observation that Abeta is elevated in AD patients and that the disease is ultimately characterized by the central deposition of insoluble senile plaques. More recent evidence, however, suggests that the presence or absence of plaque is insufficient to fully account for the deleterious role of elevated Abeta in AD. Such studies support the basis for an alternate interpretation of the Abeta cascade hypothesis. Namely, that soluble oligomers of Abeta (i.e., ADDLs) accumulate and cause functional deficits prior to overt neuronal cell death or plaque deposition. Accordingly, the following review focuses on research describing the preparation and functional activity of ADDLs in vitro and in vivo. These studies provide the basis for an alternate, ADDL-based, view of the Abeta cascade hypothesis and accounts for the disconnect between plaque burden and cognitive deficits. Possible therapeutic approaches aimed at lowering ADDLs in AD patients are also considered. PMID- 16712495 TI - Genetically modified mice models for Alzheimer's disease. AB - Transgenic mice models for Alzheimer's disease (AD) are essential to the understanding of disease pathophysiology, develop robust behavioral models and predict outcomes from pharmacological interventions. In the last 10 years, numerous mice models have been developed particularly focusing on the amyloid precursor protein-processing pathway and Tau pathology since brain amyloid deposits and Tau tangles are some of the primary neuropathological consequences of AD. Current views on the amyloid hypothesis and mice models relating to the role of soluble Abeta oligomers and intracellular Abeta in AD pathophysiology will be reviewed. Several novel transgenic mice models that have recently been developed and their potential impact on understanding disease pathogenesis will also be summarized. PMID- 16712496 TI - The role of MRI and PET/SPECT in Alzheimer's disease. AB - Alzheimer's disease (AD) is difficult to diagnose in its early stages, and even if detected early, there is no preventative treatment. Imaging modalities such as MRI, PET, and SPECT have the potential to contribute to both the diagnosis of Alzheimer's disease, as well as assist in the search for more effective treatments. A number of AD-related biomarkers have been proposed and evaluated. The use of PET imaging to detect alterations in regional brain metabolism using [(18)F]FDG has enabled more sensitive and accurate early diagnosis of AD, especially in conjunction with traditional medical evaluation. Additionally, magnetic resonance imaging and spectroscopy provide a wide range of biomarkers that have been shown to correlate with the progression of AD. Some of these markers have been pursued in clinical trials. Progress has been made toward the evaluation of other more AD-specific biomarkers. However, many questions remain concerning the validity and sensitivity of these imaging biomarkers to aid in the assessment of potential new treatments, especially those related to increased levels of amyloid peptides in the brain. PMID- 16712505 TI - Unfolding during urea denaturation of a low molecular weight phytocystatin (thiol protease inhibitor) purified from Phaseolus mungo (Urd). AB - In the present study, two phytocystatins were purified to homogeneity as peaks I and II with molecular weights of 19 kDa and 17 kDa, respectively, as determined by SDS-PAGE and mass spectrometry. Both PMCs I and II were purified with a greater than 1000-fold purification and overall yield of about 16-18%. The effect of urea on PMC I and II was analysed by fluorescence and Circular Dichroism (CD) spectroscopy. Fluorescence studies suggest a red shift of the maximum emission at higher urea concentrations. PMC I and II are extremely stable protein inhibitors with regards to temperature and pH stability. FTIR studies show predominant alpha helical structure in both the cystatins. CD analysis results show change in urea concentration-dependent loss in ellipticity, as well as in the shape of the CD spectrum compared to the intact phytocystatin. PMID- 16712506 TI - Cyclization of alpha-synuclein derived peptide increases its chaperone-like activity. AB - We have analyzed a series of peptides derived from the C-terminus of alpha synuclein for chaperone-like activity. Specifically, a cyclic peptide generated by introducing a disulfide bond was observed to increase chaperone-like activity. This is the first example of a disulfide-crosslinked peptide that exhibits activity against protein aggregation and activity loss. PMID- 16712507 TI - NMR characterization of recombinant transmembrane protein CB2 fragment CB2(180 233). AB - The expression of membrane proteins has been the bottleneck for their structural studies. Recently, we developed a method to obtain milligram quantities of isotope-labeled seven transmembrane G-protein coupled cannabinoid (CB) receptor fragment in E. coli. In order to verify this method and confirm the recombinant isotope-labeled CB2 fragment, 3D hetero-nuclear NMR techniques were used to analyze the structure of the fragment CB2(180-233) in DMSO-d6 solvent. The sequential assignments of TM5 and intra-cellular loop 3 were accomplished, which confirmed the experimental protocols of isotope-labeled recombinant protein expression, fusion protein cleavage, and membrane protein purification. The obtained structure also showed alpha-helix in the TM5 region, but it was interrupted by a disordered region (Gly204_ILe206). These results further revealed that our established approach is a promising method to express recombinant membrane proteins for their structural studies. PMID- 16712508 TI - Production and purification of recombinant human glucagon overexpressed as intein fusion protein in Escherichia coli. AB - Chemico-enzymatic synthesis and cloning in Esherichia coli of an artificial gene coding human glucagon was performed. Recombinant plasmid containing hybrid glucagons gene and intein Ssp dnaB from Synechocestis sp. was designed. Expression of the obtained hybrid gene in E. coli, properties of the formed hybrid protein, and conditions of its autocatalytic cleavage leading to glucagon formation were studied. PMID- 16712509 TI - The odorant-binding protein from Canis familiaris: purification, characterization and new perspectives in biohazard assessment. AB - In this report we show the purification to homogeneity and a partial characterization of a new odorant-binding protein from Canis familiar (CfOBP) nasal mucosa. In addition, we report preliminary data on the utilization of CfOBP as a probe for the development of a refractive index-based biosensor. PMID- 16712510 TI - Expression of active alpha-N-acetylglucosaminidase/TAT Chimerae in cultured Spodoptera frugiperda cells. AB - We examined the production and secretion of fusion constructs containing alpha-N acetylglucosaminidase, the enzyme deficient in Sanfilippo B, and either wildtype TAT or modified TAT in cultured Spodoptera frugiperda cells. All constructs exhibited successful expression of active enzyme, suggesting the future possibility of utilizing TAT/alpha-N-acetylglucosaminidase chimerae in enzyme replacement therapy. PMID- 16712511 TI - Tyrosine sulfation of arylsulfatase A and its peptide. AB - Purified human liver arylsulfatase A (ASA) as well as an ASA peptide (residues 28 39) were sulfated by tyrosyl protein sulfotransferase in vitro. The media, but not the cell lysate, of normal human fibroblasts contained a tyrosine sulfated protein (pI = 4.5-5.5). This protein was not present in either media or cell lysate of human fibroblasts lacking ASA protein. These results suggest that tyrosine sulfation facilitates secretion of ASA and that this may have pathophysiological consequences. PMID- 16712512 TI - Purification, and properties of a bovine uricase. AB - Uricase from bovine kidney, purified to homogeneity level, had a molecular weight of 70 kDa. The apparent K(m) and V(max) values for uric acid hydrolysis were 0.125 mM and 102 IU mg(-1) protein respectively. The activation energy requirement for uric acid hydrolysis by uricase and inactivation of enzyme were 11.6 and 14.5 kJ/M respectively. Both enthalpy (Delta H*) and entropy of activation (Delta S*) for uricase activity were lower than those reported for some thermostable enzymes. PMID- 16712513 TI - Expression, purification, and characterization of an enzymatically active truncated human rho-kinase I (ROCK I) domain expressed in Sf-9 insect cells. AB - Rho Kinase I (ROCK I) is a serine/threonine kinase that is involved in diverse cellular signaling. To further understand the physiological role of ROCK I and to identify and develop potent and selective inhibitors of ROCK I, we have overexpressed and purified a constitutively active dimeric human ROCK I (3-543) kinase domain using the Sf9-baculovirus expression system. In addition, using a limited proteolysis technique, we have identified a minimal functional subdomain of ROCK I that can be used in crystallization studies. The availability of multimilligram amounts of purified and well characterized functional human ROCK I kinase domains will be useful in screening and structural studies. PMID- 16712514 TI - Fate of influenza A virus proteins. AB - In this review we summarize the current state, history, future, mutation tendency and species susceptibility of influenza A virus proteins based on our probabilistic analyses on amino acid pairs, and compare the current state of influenza A virus proteins with that of proteins which we have studied in the past. PMID- 16712515 TI - The early events of alpha-synuclein oligomerization revealed by photo-induced cross-linking. AB - Assembly of alpha-synuclein (alpha-Syn) into neurotoxic oligomers and fibrils is an important pathogenic feature of Parkinson's disease. Studying the early events of alpha-Syn aggregation, such as oligomerization and nucleation, is indispensable to understanding the complicated process. Here, photo-induced cross linking of unmodified proteins (PICUP) technique is applied to elucidate the early-stage oligomerization of alpha-Syn. Results show that alpha-Syn in solution exhibits a mixture of various species, including at least monomers, dimers and trimers. Aggregation of alpha-Syn probably originates from the dimeric and trimeric seeds. Furthermore, the N-terminal amphipathic region is proposed to be required for the oligomerization (dimerization and trimerization) process. This observation may extend our knowledge on the early events of alpha-Syn aggregation and the neurotoxic aggregation species. PMID- 16712516 TI - Network analysis of the protein chain tertiary structures of heterocomplexes. AB - In this paper, the tertiary structures of protein chains of heterocomplexes were mapped to 2D networks; based on the mapping approach, statistical properties of these networks were systematically studied. Firstly, our experimental results confirmed that the networks derived from protein structures possess small-world properties. Secondly, an interesting relationship between network average degree and the network size was discovered, which was quantified as an empirical function enabling us to estimate the number of residue contacts of the protein chains accurately. Thirdly, by analyzing the average clustering coefficient for nodes having the same degree in the network, it was found that the architectures of the networks and protein structures analyzed are hierarchically organized. Finally, network motifs were detected in the networks which are believed to determine the family or superfamily the networks belong to. The study of protein structures with the new perspective might shed some light on understanding the underlying laws of evolution, function and structures of proteins, and therefore would be complementary to other currently existing methods. PMID- 16712517 TI - Leukocyte function-associated antigen-1: structure, function and application prospects. AB - This review focuses on the structure, function and pathological role of leukocyte function-associated antigen-1 (LFA-1) which is a heterodimeric protein consisting of two subunits. LFA-1 plays a most important role in the immune system including adhesion, extravasation, migration, apoptosis, cytotoxicity, cytokine production, and proliferation of lymphocytes. Therefore, T-cell activation can be suppressed by blocking ICAM-1/LFA-1 interaction in autoimmune diseases and organ transplantation. Many different inhibitors (i.e. antibodies, peptides, small molecules) have been demonstrated to block ICAM-1/LFA-1 interaction, and some of them are promising for medical treatment or have reached clinical trials. PMID- 16712518 TI - Comparative study of apoptosis induced by H(2)O(2) and NO: limitation of apoptosis induced by NO due to slower recovery of activity of NO-modified caspase. AB - Both H(2)O(2) and NO can act as apoptogens, triggering apoptosis in many cells. They are also well known inhibitors of caspases, essential enzymes in apoptosis. The differences between these two agents as apoptosis inducers and how caspases mediate apoptosis with these inhibitory agents is still unclear. Consistent with the previous reports, these two agents induced apoptosis accompanied by caspase activation with limitation of all apoptotic events for NO. It was found that NO modified caspase-3 showed a slower recovery of its activity in the presence of the reducing agents compared to that of H(2)O(2) modification. This is one possible cause of the limited apoptosis in the case of NO. PMID- 16712519 TI - Purification and characterization of arginine kinase from locust. AB - L-Arginine kinase (AK; ATP:L-arginine N-phosphotransferase; EC 2.7.3.3) catalyzes the reversible transphosphorylation between N-phospho-L-arginine (PArg) and ATP thus buffering cellular ATP levels. AK was purified from the leg muscle of the locust Migratoria manilensis by Sephacryl S-200 HR gel filtration chromatography and DEAE Sepharose CL-6B fast flow anion exchange chromatography to an apparent homogeneity with a recovery of 80%. The enzyme behaved as monomeric protein with molecular mass of about 40 kD, and had a pH and temperature optimum of 8.6 and 30 degrees C, respectively, and a pI of about 6.3. The Michaelis constants for synthesis of PArg are 0.936 and 1.290 mM for L-arginine and ATP, respectively and k(cat)/K(m)(Arg) 174. The activity of AK required divalent cations such as Mg(2+) and Mn(2+). In the presence of Cu(2+) and Zn(2+), AK activity was greatly inhibited. The intrinsic protein fluorescence emission maximum at 330 nm using the excitation wavelength at 295 nm suggested that tryptophan residues are below the surface of the protein and not exposed to solvent. PMID- 16712520 TI - Purification and properties of laticeptin, an antimicrobial peptide from skin secretions of the South American frog Leptodactylus laticeps. AB - Norepinephrine-stimulated skin secretions from the Sante Fe frog Leptodactylus laticeps contained high concentrations of a peptide, termed laticeptin, with the primary structure Gly-Val-Val-Asp-Ile-Leu-Lys-Gly-Ala-Ala-Lys-Asp-Leu-Ala-Gly-His Leu-Ala-Thr-Lys-Val-Met-Asn-Lys-Leu.NH(2). Laticeptin inhibited the growth of selected Gram-negative bacteria but the lack of activity against Gram-positive bacteria and the very low hemolytic activity is probably a consequence of the weak amphipathicity of the peptide in its alpha-helical conformation. PMID- 16712521 TI - Crystallization and preliminary X-ray crystallographic analysis of yeast tyrosyl tRNA synthetase complexed with its cognate tRNA. AB - Yeast tyrosyl-tRNA synthetase (yTyrRS) has been crystallized by the vapor diffusion method in the presence of its cognate tRNA(Tyr). The crystals belong to a tetragonal space group P4(1)2(1)2 with cell dimensions of a = b = 63.85 Angstrom, and c = 330.3 Angstrom. The asymmetric unit contains one molecule each of yTyrRS and tRNA(Tyr) (one-half of a 2:2 complex). X-ray diffraction data have been collected up to 2.5 Angstrom resolution. PMID- 16712522 TI - Selection and characterization of an HIV-1 gp120-binding affibody ligand. AB - To evaluate the possibility of generating novel proteins binding to highly glycosylated viral proteins, affibody ligands were selected by bacteriophage display technology to the HIV-1 envelope glycoprotein gp120 (glycoprotein 120), from a combinatorial protein library based on the 58-amino-acid-residue staphylococcal Protein A domain. The predominant variant from the bacteriophage selection was produced in Escherichia coli and characterized by biosensor analyses. Both univalent and bivalent affibody molecules were shown to bind selectively to the gp120 target molecule in a biosensor analysis. The dissociation equilibrium constants (KD) were determined to be approx. 100 nM for the univalent affibody and 10 nM for the bivalent affibody, confirming the stronger gp120 binding of the bivalent affibody ligand. The affibody constructs were further introduced into the Ad5 (adenovirus type 5) fibre gene, and the recombinant fibres were shown to bind selectively to gp120 in a biosensor analysis and to gp160 transiently expressed in African-green-monkey (Cercopithecus aethiops) kidney cells. Neither the affibody ligand nor the Ad5 fibres showed any virus neutralization activity, suggesting that the affibody bound to a non-neutralizing site on gp120. To investigate the binding site for the affibody ligand on gp120, CD4 (cluster of differentiation 4) and a panel of mAbs (monoclonal antibodies) known to bind to gp120 were allowed to compete with the affibody ligand in a biosensor study. Two mAbs, 670-30D and 697-30D, were found to compete with gp120 for overlapping binding sites. Although neutralization effects were not achieved in this initial investigation, the successful selection of a gp120-binding affibody ligand indicates that future affibody-based strategies might evolve to complement antibody-based efforts for HIV-1 therapy. Strategies for directed selection of affibody ligands binding to neutralizing epitopes and the potential of using adenovirus for gene-therapy mediated efforts are discussed. PMID- 16712523 TI - Involvement of the SMRT/NCoR-HDAC3 complex in transcriptional repression by the CNOT2 subunit of the human Ccr4-Not complex. AB - In eukaryotic cells, the Ccr4-Not complex can regulate mRNA metabolism at various levels. Previously, we showed that promoter targeting of the CNOT2 subunit resulted in strong repression of RNA polymerase II transcription, which was sensitive to the HDAC (histone deacetylase) inhibitor, trichostatin A [Zwartjes, Jayne, van den Berg and Timmers (2004) J. Biol. Chem. 279, 10848-10854]. In the present study, the cofactor requirement for CNOT2-mediated repression was investigated. We found that coexpression of SMRT (silencing mediator for retinoic acid receptor and thyroid-hormone receptor) or NCoR (nuclear hormone receptor co repressor) in combination with HDAC3 (or HDAC5 and HDAC6) augmented the repression by CNOT2. This repressive effect is mediated by the conserved Not-Box, which resides at the C-terminus of CNOT2 proteins. We observed physical interactions of CNOT2 with several subunits of the SMRT/NCoR-HDAC3 complex. Our results show that the SMRT/NCoR-HDAC3 complex is a cofactor of CNOT2-mediated repression and suggest that transcriptional regulation by the Ccr4-Not complex involves regulation of chromatin modification. PMID- 16712524 TI - Body composition and the apoB/apoA-I ratio in migrant Asian Indians and white Caucasians in Canada. AB - Migrant and native South Asians appear to be at increased risk of Type II diabetes mellitus and coronary disease. The aim of the present study was to determine the relationship between the most accurate summary index of the lipoprotein-related risk of vascular disease, the apoB (apolipoprotein B 100)/apoA-I (apolipoprotein A-I) ratio, and body composition in established migrant South Asians and white Caucasians living in Canada. Men and women living in Montreal, Canada between the ages of 20-60 years were recruited for participation in the study. Subjects were excluded if they had a history of cardiovascular disease or were taking lipid-lowering medication. Individuals identified themselves as Asian Indian or Caucasian. Anthropometric measurements were collected, including weight, height, waist circumference, hip circumference and body fat percentage. Plasma samples were analysed for total cholesterol, HDL C (high-density lipoprotein-cholesterol), apoA-I and apoB. Indian subjects had a substantially higher WHR (waist-to-hip ratio) than Caucasian subjects [men, 0.93+/-0.01 compared with 0.86+/-0.01 respectively (P<0.001); women, 0.88+/-0.01 compared with 0.77+/-0.01 respectively (P<0.0001)]. WHR correlated strongly with body fat percentage in Caucasians (men, r=0.63, P=0.0002; women, r=0.74, P<0.0001). By contrast, there was no correlation in Indians (men, r=0.22, P value not significant; women, r=0.23, P value not significant). In addition, Indian men and women had a higher apoB/A-I ratio than Caucasians [men, 0.85+/-0.04 compared with 0.66+/-0.04 respectively (P=0.001); women, 0.73+/-0.04 compared with 0.56+/ 0.03 respectively (P=0.0003)]. Of interest, there were also significant correlations between the apoB/apoA-I ratio and WHR in all of the groups, except the Indian women, which were stronger than the correlation of the apoB/apoA-I ratio with BMI. On the other hand, there was no significant relationship between the apoB/apoA-I ratio and the body fat percentage in any of the groups. In conclusion, the present study confirms that, as body fat percentage increases, the distribution of body fat differs between migrant Indians and Caucasians living in Canada. It also relates differences in body fat distribution to differences in the apoB/apoA-I ratio, providing at least part of the answer as to why South Asians may be at increased risk of vascular disease. PMID- 16712525 TI - The tert-butylhydroquinone-mediated activation of the human thioredoxin gene reveals a novel promoter structure. AB - Thioredoxin is a redox-active protein that plays multiple roles in regulating cell growth, cell signalling and apoptosis. Here, we have demonstrated that a complex mechanism involving multiple regulatory elements is involved in the tBHQ [tert-butylhydroquinone or 2,5-di-(t-butyl)-1,4-hydroquinone]-mediated activation of the thioredoxin gene. Luciferase assays, utilizing various wild-type and mutated thioredoxin promoter fragments, revealed roles for the ORE (oxidative stress responsive element), ARE (antioxidant responsive element), three Sp1 (specificity protein 1)-binding sites and the TATA box in the activation of the thioredoxin gene by tBHQ. The ORE required the presence of the ARE to elicit its response, whereas the independent removal of three Sp1-binding sites and the TATA box also decreased activation of the thioredoxin gene, with mutation of the TATA box having the greatest effect. Real-time RT (reverse transcriptase)-PCR analysis also revealed varying roles for two TSSs (transcription start sites) in the activation of the thioredoxin gene by tBHQ. Transcription was initiated from both TSSs; however, different response rates and fold inductions were observed. Together, these results suggest that the thioredoxin gene is controlled by a novel arrangement of two overlapping core promoter regions, one containing a TATA box and the other TATA-less. Altering the intracellular levels of thioredoxin in a breast cancer cell line also influenced the induction of thioredoxin transcription in response to tBHQ. Stable transfections with a redox-inactive thioredoxin mutant produced 3.6 times higher induction levels of thioredoxin transcription compared with control cells, indicating an intrinsic form of control of promoter activity by the thioredoxin system itself. PMID- 16712526 TI - Crystal structure of the SENP1 mutant C603S-SUMO complex reveals the hydrolytic mechanism of SUMO-specific protease. AB - SUMO (small ubiquitin-related modifier)-specific proteases catalyse the maturation and de-conjugation processes of the sumoylation pathway and modulate various cellular responses including nuclear metabolism and cell cycle progression. The active-site cysteine residue is conserved among all known SUMO specific proteases and is not substitutable by serine in the hydrolysis reactions demonstrated previously in yeast. We report here that the catalytic domain of human protease SENP1 (SUMO-specific protease 1) mutant SENP1C(C603S) carrying a mutation of cysteine to serine at the active site is inactive in maturation and de-conjugation reactions. To further understand the hydrolytic mechanism catalysed by SENP1, we have determined, at 2.8 A resolution (1 A = 0.1 nm), the X ray structure of SENP1C(C603S)-SUMO-1 complex. A comparison of the structure of SENP2-SUMO-1 suggests strongly that SUMO-specific proteases require a self conformational change prior to cleavage of peptide or isopeptide bond in the maturation and de-conjugation processes respectively. Moreover, analysis of the interface of SENP1 and SUMO-1 has led to the identification of four unique amino acids in SENP1 that facilitate the binding of SUMO-1. By means of an in vitro assay, we further demonstrate a novel function of SENP1 in hydrolysing the thioester linkage in E1-SUMO and E2-SUMO complexes. The results disclose a new mechanism of regulation of the sumoylation pathway by the SUMO-specific proteases. PMID- 16712529 TI - Thrombolysis for stroke: policy should be based on science, and not on politics, money or fear of malpractice. PMID- 16712527 TI - Identification and characteristics of the structural gene for the Drosophila eye colour mutant sepia, encoding PDA synthase, a member of the omega class glutathione S-transferases. AB - The eye colour mutant sepia (se1) is defective in PDA {6-acetyl-2-amino-3,7,8,9 tetrahydro-4H-pyrimido[4,5-b]-[1,4]diazepin-4-one or pyrimidodiazepine} synthase involved in the conversion of 6-PTP (2-amino-4-oxo-6-pyruvoyl-5,6,7,8 tetrahydropteridine; also known as 6-pyruvoyltetrahydropterin) into PDA, a key intermediate in drosopterin biosynthesis. However, the identity of the gene encoding this enzyme, as well as its molecular properties, have not yet been established. Here, we identify and characterize the gene encoding PDA synthase and show that it is the structural gene for sepia. Based on previously reported information [Wiederrecht, Paton and Brown (1984) J. Biol. Chem. 259, 2195-2200; Wiederrecht and Brown (1984) J. Biol. Chem. 259, 14121-14127; Andres (1945) Drosoph. Inf. Serv. 19, 45; Ingham, Pinchin, Howard and Ish-Horowicz (1985) Genetics 111, 463-486; Howard, Ingham and Rushlow (1988) Genes Dev. 2, 1037 1046], we isolated five candidate genes predicted to encode GSTs (glutathione S transferases) from the presumed sepia locus (region 66D5 on chromosome 3L). All cloned and expressed candidates exhibited relatively high thiol transferase and dehydroascorbate reductase activities and low activity towards 1-chloro-2,4 dinitrobenzene, characteristic of Omega class GSTs, whereas only CG6781 catalysed the synthesis of PDA in vitro. The molecular mass of recombinant CG6781 was estimated to be 28 kDa by SDS/PAGE and 56 kDa by gel filtration, indicating that it is a homodimer under native conditions. Sequencing of the genomic region spanning CG6781 revealed that the se1 allele has a frameshift mutation from 'AAGAA' to 'GTG' at nt 190-194, and that this generates a premature stop codon. Expression of the CG6781 open reading frame in an se1 background rescued the eye colour defect as well as PDA synthase activity and drosopterins content. The extent of rescue was dependent on the dosage of transgenic CG6781. In conclusion, we have discovered a new catalytic activity for an Omega class GST and that CG6781 is the structural gene for sepia which encodes PDA synthase. PMID- 16712530 TI - Evolution of emergency medicine teaching for medical students. PMID- 16712531 TI - Paramedic intubation of patients with severe head injury: a review of current Australian practice and recommendations for change. AB - Secondary brain injury may occur early after severe traumatic brain injury due to hypoxia and/or hypotension. Prehospital care by ambulance paramedics has the goal of preventing and treating these complications and, thus, improving outcomes. In Australia, most ambulance services recommend paramedics attempt endotracheal intubation in patients with severe head injury. Even though most patients with severe head injury retain airway reflexes, most states do not allow the use of appropriate drugs to facilitate intubation. In contrast, recent evidence from trauma registries suggests that this approach may be associated with significantly worse outcomes compared with no intubation. Two states allow intubation facilitated by sedative (but not relaxant) drugs, but this has a low success rate and could worsen brain injury because of a decrease in cerebral perfusion pressure. For road-based paramedics, the role of rapid sequence intubation is uncertain. Given the risks of this procedure and the lack of proven benefit, this procedure should not be introduced without supportive evidence from randomised, controlled trials. In contrast, for safety reasons, comatose patients transported by helicopter should undergo rapid sequence intubation prior to flight. However, this is not authorised in most states, despite good supportive evidence that this can be safely and effectively undertaken by paramedics. Finally, there is evidence that inadvertent hyperventilation is associated with adverse outcome, yet only two ambulance services use waveform capnography in head injury patients who are intubated. Overall, current paramedic airway practice in most states of Australia is not supported by the evidence and is probably associated with worse patient outcomes after severe head injury. For road-based paramedics, rapid transport to hospital without intubation should be regarded as the current standard of care. Rapid sequence intubation should be limited to use within appropriate clinical trials, or patients transported by helicopter. For patients who are intubated, waveform capnography is essential to confirm tracheal placement and to prevent inadvertent hyperventilation. PMID- 16712532 TI - Fingertip injuries. AB - Fingertip injuries are a potential cause of significant morbidity within the workplace and the community. Although they account for a low proportion of ED presentations, importantly they can frequently be managed entirely by emergency staff. A literature review was undertaken to identify the epidemiology, the assessment process and the management options for fingertip injuries as relevant to emergency medicine. PMID- 16712533 TI - Head computed tomography guidelines are being followed. AB - OBJECTIVE: Several guidelines have been developed to direct the ordering of head computed tomography (CT) for patients, but most are clinical presentation specific. Recently, an integrated guideline for ordering emergent head CT for patients who present to the ED of Western Hospital, Footscray, Victoria, Australia, was developed in response to concerns raised regarding perceived over utilization of head CT for ED patients. Our aim was to determine compliance with the guideline. METHODS: This was an explicit retrospective medical record review of patients who presented to the ED of Western Hospital between 1/04/2004 and 17/6/2004 and had a head CT as part of their assessment. Clinical information for these cases was compared with guideline recommendations. Data are described by descriptive statistics. RESULTS: Of the 231 cases that were included in the study, 65 (28.1%, 95% confidence interval 23-35%) had abnormal CT findings. Guidelines were adhered to in 217 (93.9%, 95% confidence interval 91-97%) cases. For the cases where the guidelines were not adhered to (14; [6.1%]), there was only one abnormal scan the clinical significance of which is not clear. CONCLUSION: The study found that compliance with head CT guideline was high. This suggests that the guideline is both clinically relevant and supported by ED doctors or conversely that the guideline is concordant with existing ordering practices of the ED. PMID- 16712534 TI - NexTemp thermometer can be used interchangeably with tympanic or mercury thermometers for emergency department use. AB - OBJECTIVES: To determine the agreement between the chemical dot NexTemp thermometer with mercury and tympanic thermometers and the repeatability of measurements using these devices. METHODS: A prospective study involving a convenience sample of 194 consenting adult patients presenting to the ED, Freemasons Hospital, East Melbourne, Victoria, Australia. A survey of emergency medical staff was conducted to determine what they considered an acceptable level of agreement and repeatability for a putative new thermometer. The NexTemp thermometer's performance was judged against this. For each thermometer, a set of two temperature measurements was made in every patient. The sequence of the set of readings (and hence device) was random between patients and the staff member performing one set was blinded to the results of the other two sets of readings in each patient. The method of Bland and Altman was used for assessing agreement and repeatability. RESULTS: Clinicians considered that a new thermometer should exhibit repeatability of +/- 0.3 degrees C and agree with existing devices within +/- 0.5 degrees C. The tympanic thermometer had 95% limits of repeatability of 0.8-0.5 degrees C compared with the NexTemp (-0.3-0.4 degrees C) and mercury thermometers (-0.3-0.4 degrees C). The NexTemp thermometer agreed with mercury thermometer within -0.6-0.5 degrees C. The tympanic thermometer agreed with the mercury thermometer within -1.0-1.1 degrees C. CONCLUSION: Based on temperature measurement only, the NexTemp thermometer can be used interchangeably with current mercury and tympanic thermometers. PMID- 16712535 TI - Investigation of the variables that impact upon the knowledge of cardiac risk factors. AB - OBJECTIVE: Awareness of cardiac disease risk factors is required before they can be modified. The present study aimed to investigate risk factor knowledge and the variables that impact upon this knowledge. METHODS: We undertook an analytical, cross-sectional survey of 226 patients attending an ED. Patients were asked to recall as many cardiac risk factors as possible and to rate the risk of nine given risk factors. Data relating to sources of risk factor information were collected. Uni- and multivariate (multiple linear regression) analyses determined variables that impacted upon the risk factor knowledge score. RESULTS: Mean patient age was 60.2 +/- 15 years, 55.3% (95% confidence interval 48.6-61.9) were male and 19.9% (95% confidence interval 15.0-25.8) had known cardiac disease. The mean risk factor knowledge score of 2.5 +/- 1.5 out of a possible 12 (median 3) was indicative of poor knowledge. Smoking, poor diet and stress/worry/tension were the most common factors reported. Variables impacting significantly on the knowledge score were English as a first language (P < 0.001), age (negative correlation, P < 0.001) and the receipt of information relating to cardiac health (P < 0.001). The patients' general practitioner and the media were the most important sources of information. CONCLUSIONS: Patients' knowledge of cardiac risk factors is generally poor and education strategies are indicated. At the individual level, the general practitioner is likely to remain as an important influence on knowledge. However, complementary media education programs are indicated at the community level. Patient subgroups at 'high risk' through poor knowledge should be specifically targeted. PMID- 16712537 TI - Procedural skills quality assurance among Australasian College for Emergency Medicine fellows and trainees. AB - OBJECTIVE: Presently, no objective quality control mechanism exists for monitoring procedural skills among Australasian College for Emergency Medicine trainees. The present study examined trainee and fellow procedural experience and perceived competency, participation in accredited training courses and support for a procedural logbook. METHODS: A cross-sectional mail survey of Australasian College for Emergency Medicine advanced trainees and fellows was performed. Experience and perceived competency in 23 common and important ED procedures were examined. RESULTS: In total, 202 fellows and 264 trainees responded (overall response rate 39.0%). Overall, fellow procedural experience and perceived competency were reasonable. However, some fellows had never performed a number of procedures including some common procedures (e.g. nasal packing, fracture reduction) and there were reports of 'very poor' competency for 17 (73.9%) procedures. Trainee experience and perceived competency were less than the fellows but showed similar patterns. Perceived numbers of each procedure required to achieve competency varied considerably between the procedures and among the respondents. However, there were no significant differences in the perceived numbers reported by the trainees and the fellows (P > 0.05). Variable proportions of trainees and fellows had undertaken courses that incorporated procedural skills training. More fellows (75.7%, 95% confidence interval 69.1-81.4) than trainees (59.9%, 95% confidence interval 53.6-65.8) supported the use of a procedural logbook (P = 0.003). CONCLUSIONS: Lack of experience in some procedures among some fellows, especially commonly performed procedures, might represent a deficiency in existing quality assurance mechanisms for procedural skills training. Greater participation in skills courses, to improve experience in difficult and uncommonly encountered procedures, is recommended. Improved quality assurance mechanisms, including a procedural logbook, should be considered. PMID- 16712536 TI - Sutured wounds: factors associated with patient-rated cosmetic scores. AB - OBJECTIVE: To determine the association between wound characteristics, wound management in the ED and patient-rated cosmetic appearance of sutured wounds. Our hypothesis was that practitioner seniority would most strongly predict outcome. METHODS: Prospective recruitment of patients with lacerations sutured at the primary ED visit was performed. Data collected included patient demographics, wound characteristics and wound management. A standardized telephone questionnaire was administered 14 days and 3 months later. Scar appearance was scored using a verbal rating scale from zero to 10. Data were obtained about suture removal, antibiotic compliance, infection and dehiscence rates at 2 weeks. Associations of variables with cosmetic scores were analysed using multivariate linear regression. RESULTS: One hundred and thirty-two patients were evaluated. Mean cosmetic scores were not significantly associated with seniority (P = 0.07). Lacerations repaired by senior practitioners were more likely to result from glass or general trauma (P = 0.007), be shorter (P = 0.03), be cleaned with antiseptic (P = 0.03), not to re-open (P = 0.01) or require re-suturing (P = 0.03). Following multivariate regression factors significantly associated with cosmetic scores at 14 days and 3 months were site of injury (P < 0.003) and time from injury to repair (P < 0.009). Wounds of the torso, leg or foot had lower cosmetic scores at both time-points. An association with age (P = 0.04) was present at 3 months. CONCLUSIONS: Previous research found improvement between short-term doctor-rated cosmesis and training beyond internship. Our study demonstrated a non-significant trend relationship between seniority and patient rated outcome, both short and long-term. However, staff seniority was overshadowed by the site of laceration and time from injury to repair. PMID- 16712538 TI - Medical student education in emergency medicine: new model from South Africa. AB - Emergency medicine is a new specialty in South Africa. Postgraduate training, degrees and diplomas have been introduced and this should make a significant difference, in due course, to emergency care in the country. The University of the Witwatersrand, Johannesburg, South Africa, like many universities in Australia, embarked upon a Graduate Entry Medical Programme. This implied significant curriculum change and gave opportunity for the development of formal emergency medicine training for students, for the first time. After considerable debate over the needs of the graduate in South Africa a new block was developed, called the Acute and Perioperative Care block. This encompasses forensic medicine, emergency medicine, trauma and anaesthetics. The training is integrated and progressive. No similar programme has been detected elsewhere. At the end of the first year there is considerable satisfaction on the part of both teachers and students and many of the students, finding the module exciting, have indicated a wish to become involved in emergency medicine in the future. The structure of the course is laid out and might be of relevance to colleagues with an interest in medical education. PMID- 16712539 TI - National Emergency Department Overcrowding Study tool is not useful in an Australian emergency department. AB - OBJECTIVE: To determine the accuracy and usefulness of the National Emergency Department Overcrowding Study (NEDOCS) tool in an urban hospital ED in Australia by direct comparison with subjective assessment by senior ED staff. METHOD: A sample of simultaneous subjective and objective data pairs were collected six times a day for a period of 3 weeks. All senior medical staff in the ED answered a brief questionnaire along with the senior charge nurse for the ED. Simultaneously, the senior charge nurse also documented the total number of patients in the ED, the number of patients awaiting admission, the number of patients on ventilators, the longest time waited by an ED patient for ward bed, and the waiting time for the last patient from the Waiting Room placed on a trolley. The objective indicators were entered into a Web-based NEDOCS tool and transformed scores were compared with the averaged and transformed subjective scores for each sample time. Bland-Altmann and Kappa statistics were used to test the agreement between the objective and subjective measuring methods. RESULTS: The mean difference between the subjective and objective methods was small (3.5 [95% confidence interval -0.875-7.878] ); however, the 95% limits of agreement was wide (-46.52-53.43). The Kappa statistic used to assess the extent of reproducibility between categorical variables was 0.31 (95% confidence interval 0.17-0.45). CONCLUSION: The present study suggests that NEDOCS method of processing the objective overcrowding data does not accurately reflect the subjective assessment of the senior staff working at that time in the ED. This might be because the assumptions of the original NEDOCS study are flawed. PMID- 16712541 TI - Refuge from the Lord's Resistance Army in Uganda: a report from a Medecins Sans Frontieres team leader. AB - In 2005 the author worked for Medecins Sans Frontieres (MSF, or Doctors without Borders) in northern Uganda. This short report shares some background information on MSF, the situation in northern Uganda as a consequence of the activities of the Lord's Resistance Army (LRA), the work of the MSF project and the personal experience of being there. PMID- 16712542 TI - Acute retrobulbar haemorrhage: an ophthalmic emergency. AB - Acute retrobulbar haemorrhage is a rare complication of blunt eye trauma. It can be sight-threatening but is reversible when recognized and treated promptly. We presented a case of acute retrobulbar haemorrhage with profound reduction of vision. Emergency surgical decompression was performed to preserve the vision. Clinical features and management are discussed. PMID- 16712540 TI - Evolution of health information management and information technology in emergency medicine. AB - The implementation of information technology will continue to have important effects on the practice of emergency medicine. Patient registration and tracking systems, telemedicine electronic order entry systems, and the advent of electronic health records are some examples of applications that influence patient care in emergency departments. We review the literature regarding information technology advances in emergency medicine and outline the important role that emergency physicians play in developing and implementing a national health information technology strategy. PMID- 16712543 TI - Tom Hamilton Oration. Developing future leaders in emergency medicine: pathways in a hazardous world. PMID- 16712545 TI - No proof HIV antibodies are caused by a retroviral infection. PMID- 16712547 TI - Correcting the holiday roadtoll myth: Christmas and Easter holiday periods are actually safer than other times of the year. PMID- 16712548 TI - Community child health in Australia: the road ahead. PMID- 16712549 TI - Evolution of non-operative management for blunt splenic trauma in children. AB - Until the late 1960s, splenectomy was routinely performed in children who had sustained blunt splenic injury. There was based on the ability to perform splenectomy without obvious consequence; the cited 90-100% mortality for splenic trauma and the possibility of delayed rupture of the spleen. In contrast, contemporary findings in immunology and surgery demonstrated that non-operative management was not only feasible but desirable in view of the potential for overwhelming post-plenectomy infection. The history of universal splenectomy following blunt splenic trauma has been reviewed and we outline the findings that have resulted in the current standard of non-operative management following blunt splenic trauma. PMID- 16712550 TI - Intussusception in the Northern Territory: the incidence is low in Aboriginal and Torres Strait Islander children. AB - AIM: To describe the epidemiology, immunisation status and management of children with intussusception in the Northern Territory (NT), 1993-2003. METHODS: Intussusception data were obtained from all NT hospitals using the International Classification of Diseases (ICD 9/10) codes for children under 18 years of age between 1993 and 2003. Medical records of these children were used to collect information on demographics, admission date, clinical symptoms, signs and management. Immunisation data were obtained from the NT immunisation register. The NT mortality database was reviewed for deaths from intussusception in children between 1993 and 2003. One death in an Aboriginal and Torres Strait Islander child was found in the NT mortality database. Medical records for this child were destroyed and so the case definition for intussusception used in this study was not fulfilled and the child was excluded. RESULTS: Intussusception proven by radiological or surgical means was identified in 23 children from hospital records. The incidence for children with intussusception in NT is 0.65/1000 live births. The incidence of intussusception was lower in Aboriginal and Torres Strait Islander children (0.16/1000 live births) than in non Aboriginal and Torres Strait Islander children (0.92/1000 live births) (P < 0.01). CONCLUSION: The incidence of intussusception in the NT is similar to other developed countries but Aboriginal and Torres Strait Islander children have a very low incidence of intussusception. Intussusception is a rare event in the NT and will require a sensitive surveillance system to detect any potential increased risk of intussusception after the introduction of a new rotavirus vaccine. PMID- 16712552 TI - Capsule endoscopy: a new technology for investigating obscure gastrointestinal bleeding. AB - In Australia, Medicare rebate was approved in May 2005 for use of capsule endoscopy in children aged 10 years and over. Approval was granted for an interim period to allow investigation of obscure ongoing or recurrent gastrointestinal bleeding, for which no cause could be found after investigation, including conventional radiology and endoscopy. Strict eligibility criteria for Medicare rebate for the procedure have been developed. These criteria relate to patient selection, type of device, timing of the procedure, qualifications of providers and requirements for prospectively monitoring the use, uefulness, safety and cost effectiveness of the procedure in children. PMID- 16712551 TI - Adrenomedullin and total nitrite levels in children with familial Mediterranean fever. AB - AIM: Familial Mediterranean fever (FMF) is the most frequent periodic syndrome characterised by recurrent attacks of polyserositis. However, recent studies revealed that there might be an ongoing subclinical inflammation between the attacks. As nitric oxide (NO) and adrenomedullin (AM) are both synthesised in the endothelium, and mediates many functions within immune system, we considered them to be an interesting target of investigation in FMF. METHODS: Fifteen children with FMF receiving regular colchicine, ranging in age from 3 to 16 years, were investigated in comparison with 15 healthy age- and sex-matched controls. The mean age of the patients was 9.7 +/- 3.9 years. Total nitrite, a stable product of NO, was quantitated by means of the Griess reaction, while AM was measured by HPLC. RESULTS: Plasma-urinary AM and total nitrite levels were significantly higher in children with FMF. Plasma AM levels (pmol/mL) in patients and controls were 40.95 +/- 5.99 vs. 34.86 +/- 5.24, P < 0.05, and urinary AM excretion (pmol/mg creatinine) was 51.16 +/- 28.15 vs. 37.5 +/- 24.26, P < 0.05 respectively. Plasma total nitrite levels (micromol/L) in patients and controls were 44.80 +/- 10.36 vs. 32.13 +/- 9.28, P < 0.05, and urinary nitrite excretion (micromol/mg creatinine) was 2.24 +/- 1.71 vs. 1.09 +/- 0.96, P < 0.05 respectively. CONCLUSION: This study considered that AM and NO may have a role in the immuno-inflammatory process of FMF, although, whether these act to preserve, or protect against, further inflammatory injury is not clear. Our results further supports the hypothesis that these patients have subclinical inflammation between attacks. PMID- 16712553 TI - Predicting respiratory syncytial virus hospitalisation in Australian children. AB - BACKGROUND: There is limited information on respiratory syncytial virus infections among Australians, particularly those of Indigenous descent. AIM: This study identifies groups of infants at risk of hospitalisation with respiratory syncytial virus-positive lower respiratory tract infection who may be targeted for prevention with palivizumab. METHODS: Case control study: the case notes of 271 children with cases of respiratory syncytial virus-positive lower respiratory tract infection admitted to The Townsville Hospital were studied for risk factors. Controls were chosen randomly from babies born in The Townsville Hospital during that period. Multiple logistic regression analysis and classification and regression tree analysis were used to identify risk factors. RESULTS: Multiple logistic regression analysis identified birthweight <2500 g, maternal parity and marital status to be independent predictors of hospitalisation with respiratory syncytial virus-positive lower respiratory tract infection. Classification and regression tree analysis identified babies born weighing <2500 g who possessed older siblings to be at highest risk. Single mothers and smoking were additional risk factors. Indigenous babies were significantly more likely to be exposed to all of the identified risk factors. CONCLUSION: Babies born weighing <2500 g (especially with siblings) could be targeted for prevention. All Indigenous babies should be considered at high risk because of their exposure to multiple risk factors. PMID- 16712554 TI - Cost-effectiveness of respiratory syncytial virus prophylaxis with palivizumab. AB - BACKGROUND: A monoclonal antibody, palivizumab, directed against respiratory syncytial virus (RSV) has been shown to decrease hospitalisation rates. Because of its expense, the cost-effectiveness of this agent should be determined for high-risk groups. AIM: To determine characteristics of RSV infection in Townsville and the economic feasibility of palivizumab immunoprophylaxis in high risk groups. METHODS: Cases of RSV-positive bronchiolitis were retrospectively identified. Cases were grouped according to recognised risk factors. The hypothetical costs of palivizumab immunoprophylaxis for infants at risk were calculated. RESULTS: The rate of hospitalisation with RSV-positive lower respiratory tract infection was 22 per 1000 live births but increased to 50 per 1000 among Indigenous babies born weighing <2500 g. The cost of preventing an admission in each of the identified high-risk groups, based on drug costs alone, ranged from AD 69,861 to AD 88,547. CONCLUSION: Palivizumab was not cost effective in the prophylaxis of RSV in the high-risk group of infants tested here. PMID- 16712555 TI - Cytomegalovirus-associated protracted diarrhoea in an immunocompetent boy. AB - AIM: Cytomegalvirus (CMV) gastroenteritis is rarely described in immunocompetent children. We describe here an immunocompetent 1-year-old boy with CMV-associated protracted diarrhoea and discuss the pathogenesis of CMV infection of the gastrointestinal tract. METHODS: Immunohistochemical study was performed using a gastric and duodenal tissue of this patient. Furthermore, the detection of CMV DNA in stool and urine samples was performed by polymerase chain reaction method. RESULTS: CMV infection was proven by the detection of CMV-DNA in stool and urine samples and CMV-positive cells in the immunoperoxidase preparation from gastroduodenal biopsy. The biopsy specimens showed severe chronic active gastroduodenitis with numerous CD3+ T cells and CD68+ macrophages. CONCLUSIONS: We speculated from these findings that CMV invasion of the mucosal layer induced an intense immune response characterised by massive infiltration of activated T cells and macrophages in this patient. Although the pathogenesis of CMV associated gastroenteritis in immunocompetent hosts remains generally unknown, immune mechanism may play some roles in generating the chronic inflammatory lesions. PMID- 16712556 TI - Seizures, ataxia, developmental delay and the general paediatrician: glucose transporter 1 deficiency syndrome. AB - AIM: Glucose transporter 1 deficiency syndrome (GLUT1-DS) is an important condition for the general paediatrician's differential armamentarium. We describe a case series of eight patients in order to raise awareness of this treatable neurometabolic condition. The diagnosis of GLUT1-DS is suggested by a decreased absolute cerebrospinal fluid (CSF) glucose value (<2.2 mmol/L) or lowered CSF: plasma glucose ratio (<0.4). METHODS: This is a review of eight Queensland patients with GLUT1-DS. The clinical presentation, clinical course, laboratory investigations and treatment outcomes are discussed. RESULTS: The clinical features noted in our patient cohort include combinations of ataxia, developmental delay and a severe seizure disorder that is refractory to anticonvulsant medications. Seizures are the most common clinical manifestation and may be exacerbated by phenobarbitone. The paired CSF: plasma glucose results ranged from 0.2 to 0.39 (normal <0.6) with an average of 0.33. 3-O-Methyl-D Glucose uptake and GLUT1 Genotyping analysis have been performed on five patients thus far. Rapid and impressive seizure control was observed in 100% of our patients once the ketogenic diet was instituted, with half of the cohort being able to wean completely from anticonvulsants. CONCLUSION: Children presenting with a clinical phenotype consisting of a refractory seizure disorder, ataxia and developmental delay should prompt the consideration of Glucose transporter 1 deficiency syndrome. While the diagnostic test of lumbar puncture is an invasive manoeuvre, the diagnosis provides a viable treatment option, the ketogenic diet. GLUT1-DS displays clinical heterogeneity, but the value of early diagnosis and treatment is demonstrated by our patient cohort. PMID- 16712557 TI - Hepatitis B vaccination is effective for babies weighing less than 1800 g. AB - AIM: This trial studied the effectiveness of early hepatitis B (HepB) immunisation in babies weighing less than 1800 grams, born of HepB surface antigen-negative mothers. METHODS: The first vaccine dose was given once clinical stability was achieved, with second and third doses given 1 and 6 months later, respectively. HepB serology, done using Abbott ElA (phase 1) and Abbott Axsym (phase 2) before and after June 2001, respectively, was checked at birth (Sero1), prior to (Sero2) and 6 months after (Sero3) the third dose. A booster dose was recommended when Sero3 showed a non-immune status (< 10 mIU/mL). RESULTS: Median birth weight and gestational age (n = 118) were 1295 [range 475, 1780] g and 31 [range 24, 37] completed weeks, respectively. Sero1 (median age of 4 [range 1, 34] days) showed 64% (n = 113) to be non-immune. The first dose of vaccine was administered at a median weight of 1268 [range 530, 1790] g, median age of 6 [range 1-63] days and median post-menstrual age of 32 [range 24-37] completed weeks. Sero2 (median age of 179 [range 112-260] days), for 110 babies (93.2%) showed immunity in 48.2% (median titres--Phase 1: 26 [range 10, 150] mIU/mL; Phase 2: 34 [range 10, 1000] mIU/mL). Sero3 revealed seroprotection in 77.8% (median titres--Phase 1: 102 [range 12, 150] mIU/mL; Phase 2: 162 [range 16, 1000] mIU/mL). The more mature the bady at time of first dose, the more likely he is to achieve seroprotection (85% amongst those administered at and beyond 33 weeks; 91% among those administered at and beyond Day 10 at Sero3). CONCLUSIONS: Early HepB immunisation in infants < 1800 g can be safely recommended, with booster doses necessary at 1 year for some infants. PMID- 16712558 TI - Inflammatory measures in children with obstructive sleep apnoea. AB - AIM: To evaluate a range of inflammatory measures in children with obstructive sleep apnoea (OSA). METHODS: In total, 44 children with polysomnographically defined OSA (30 boys; mean age: 7.3 +/- 3.7 years) and 69 control subjects (44 boys; mean age: 7.6 +/- 4 years) were recruited. Controls were screened for symptoms of OSA by questionnaire at the time of elective surgery that was unrelated to the upper airway. Blood samples were analysed for C-reactive protein, and cytokines IL-1beta, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, GM-CSF, IFN-gamma and TNF-alpha. RESULTS: The majority of the children had mild OSA (32/44). Children with OSA (respiratory disturbance index 5.3 +/- 6.5 events/h) had significantly higher IFN-gamma and IL-8 levels than controls (P < 0.001 and 0.003, respectively), although correction for age, sex and body mass index reduced these differences (IFN-gammaP = 0.002, and IL-8 P = 0.051). There were no significant correlations between inflammatory measures and body mass index, respiratory disturbance index, or other sleep, desaturation, or arousal parameters including respiratory or spontaneous arousal indices, desaturation index or severity, sleep efficiency, or apnoea/hypopnoea duration in the OSA group. CONCLUSION: Children with OSA, even of mild severity, have significantly elevated IFN-gamma levels and a trend towards elevated IL-8 levels compared with asymptomatic controls, consistent with a pro-inflammatory effect of OSA. These changes seen in mild OSA may precede changes in other pro-inflammatory cytokines found in studies of adults with more severe and long-standing disease, implying a potential benefit from early disease identification and intervention. PMID- 16712559 TI - Disclosure of HIV/AIDS diagnosis to HIV-infected children in Thailand. AB - BACKGROUND: With the availability of highly active antiretroviral therapy, more HIV-infected children have lived longer. Many children are at the age that they should know the diagnosis. AIM: To identify the prevalence and patterns of disclosure of HIV/AIDS diagnosis to HIV-infected children. METHODS: A cross sectional study was conducted among 103 main care givers of HIV-infected children aged > or =6 years who received highly active antiretroviral therapy at Chiang Mai University and Sanpatong district hospitals, northern Thailand. RESULTS: One third (30.1%) of the children knew their HIV/AIDS status at an average age of 9.2 years. The care givers' understanding of 'knowing' did not always mean that the children were told the name of 'HIV' or 'AIDS'. Many of those care givers (84.3%) who reported that the child did not know their diagnosis had inaccurately explained to the child that he or she had some kind of disease such as allergy, lung, or liver disease. The most common reason for non-disclosure was the fear that disclosure might have negative psychological consequences to the child (53.4%). Almost all (88.7%) agreed that they should tell the children their diagnosis in the future but half needed health-care providers to help them at the event. CONCLUSION: There is a need for the development of disclosure guide-lines and models for health-care providers and care givers as there was a high rate of inaccurate disclosure and, in addition, care givers expressed their need for assistance from health-care providers for the future disclosure. PMID- 16712560 TI - Breastfeeding and obesity at 14 years: a cohort study. AB - AIM: To determine the influence of breastfeeding on overweight and obesity in early adolescence. METHODS: Data about breastfeeding duration, BMI of children at 14 years, and confounding variables, were collected from an ongoing longitudinal study of a birth cohort of 7776 children in Brisbane. Prevalence of overweight and obesity at 14 years was assessed according to duration of breastfeeding, with logistic regression being used to adjust for the influence of confounders. RESULTS: Data were available for 3698 children, and those not included were significantly different in age, educational level, income, race, birthweight, and small-for-gestational-age status. Breastfeeding for longer than six months was protective of obesity (OR 0.6, 95% CI 0.4, 0.96) though not of overweight. When confounding variables were considered the effect size diminished and lost statistical significance OR 0.8 (95% CI 0.5, 1.3). Breastfeeding for less than 6 months had no effect on either obesity or overweight though a trend was found for increased prevalence of overweight at 14 years with shorter periods of breastfeeding. CONCLUSION: This investigation contributes to the gathering body of evidence that breastfeeding for longer than 6 months has a modest protective effect against obesity in adolescence. PMID- 16712561 TI - Community paediatrics: soft science or firm foundations? AB - Community paediatrics strives to integrate the biology of health into the social and psychological worlds within which children grow and develop. Consumer demand for limited community paediatric clinical services is increasing and medico-legal pressures escalate professional and personal concern. Meanwhile, the profession, through training and professional support, has struggled to keep up. Research into community paediatrics and its integration into policy and clinical practice remains limited, raising the perception that it is a 'soft' science. Our viewpoint is that necessary progress in this field requires leadership, apprenticeship and research. We argue that to build firm foundations for the future requires structures to enable clinical specialisation and continuing professional development in this area. PMID- 16712562 TI - Community paediatrics in transition. PMID- 16712563 TI - Growth and development are fundamental dynamics for paediatrics. PMID- 16712565 TI - Community paediatrics and children's health: an idea whose time has come. PMID- 16712564 TI - Pragmatism and idealism in community child health. PMID- 16712566 TI - Withdrawal of neonatal mechanical ventilation against the parents' wishes. AB - Neonatologists and parents usually agree when intensive treatment should be stopped. We describe the management of two babies where there was disagreement between the parents and hospital staff, and discuss the medical, legal and ethical implications of the two cases. PMID- 16712567 TI - Subcutaneous fat necrosis of the newborn: be aware of hypercalcaemia. AB - Subcutaneous fat necrosis of the newborn is an uncommon, self-limiting panniculitis that usually occurs in full-term infants as a consequence of perinatal asphyxia. The cutaneous involvement may be associated with metabolic complications such as hypoglycaemia, thrombocytopenia, hypertriglyceridemia, anemia and hypercalcaemia. The delayed onset of hypercalcaemia, 1-6 months after the development of the skin manifestations, imposes a prolonged follow-up to avoid its acute toxic effects on cardiovascular and renal systems and the more durable metastatic calcifications. PMID- 16712570 TI - Changes for the better: transforming a mother/baby unit. PMID- 16712568 TI - Omenn syndrome: therapeutic effects of cyclosporin. AB - Omenn syndrome is a severe combined immunodeficiency with features of generalised erythroderma alopecia and evidence of Th2 inflammation (eosinophilia and raised IgE). We describe a differential effect of 2 calcineurin inhibitors, cyclosporine A (CsA) and tacrolimus, with CsA rapidly improving the erythroderma and lymphocytosis but tacrolimus having little effect. PMID- 16712571 TI - Hormone replacement therapy. How things have changed. PMID- 16712572 TI - A pleasant reflection. PMID- 16712573 TI - Low-carb diets linked to higher levels of 'bad' cholesterol. PMID- 16712575 TI - Discussing the health risks of douching. PMID- 16712574 TI - Considering contraception. What practitioners are saying when it comes to the pills, patches, rings and more. PMID- 16712576 TI - Multiple-birth children and their families. What nurses need to know. PMID- 16712577 TI - Fighting for life: a journal. The journey to health and wholeness in one small corner of Indonesia. PMID- 16712578 TI - Great leaders are made, not born. Tips for traveling the path to leadership excellence: part 2 of a four-part series. PMID- 16712579 TI - Off-label therapies for newborns. Miracles or mistakes? PMID- 16712580 TI - Responding to the newest evidence about SIDS. PMID- 16712581 TI - USDA launches searchable database of foods. PMID- 16712582 TI - Post-traumatic stress disorder and the violent loss of a child. PMID- 16712583 TI - Fast optical imaging of frontal cortex during active and passive oddball tasks. AB - This study used the high spatial and temporal resolution of the event-related optical signal (EROS) to investigate the timing of neuronal activity in frontal cortex during auditory target detection and passive oddball tasks. Activation in right middle frontal gyrus (MFG) peaked approximately 350 ms following rare target tones. This corresponded closely to the latency of the simultaneously recorded electrical P3 component. In addition, we found activation in left lateral MFG peaking at approximately 130 ms following tone onset for conditions that may have required response inhibition. These results correspond with activation patterns observed in similar fMRI studies, but provide temporal tags for the activated locations. These data may help bridge the gap between electrophysiological and hemodynamic measures of target detection and contribute to our understanding of the spatiotemporal dynamics of brain activity during target processing. PMID- 16712584 TI - Habituation of auditory evoked potentials in intracranial and extracranial recordings. AB - Effects of stimulus repetition are investigated in short-term habituation experiments. In these experiments, trains of stimuli are applied with longer intervals of no stimulation between the trains. In scalp recordings, an amplitude and latency decrease of the auditory N100 is usually observed at the beginning of the train. This contrasts to a recent finding with intracranial recordings, exhibiting an effect on N100 amplitude, but not on its latency. In the current study, P50 and N100 were simultaneously recorded intra- and extracranially in epilepsy patients. The amplitudes of P50 and N100 decreased in both recordings, whereas the P50 latency was not significantly affected. A latency decrease was revealed for the extracranially recorded N100, but not for the intracranial N100. This dissociation between the intracranial and scalp recordings might be explained by a different sensitivity of the two measurements for N100 generators. PMID- 16712585 TI - Task-dependent exogenous cuing effects depend on cue modality. AB - Task-dependent exogenous cuing effects on reaction time in detection and discrimination tasks have been ascribed to delayed withdrawal of attention in discrimination tasks. Alternatively, these differences may be due to cue-induced response inhibition in detection tasks. Unimodal and crossmodal versions of the Posner paradigm were examined with short cue-target intervals. Targets above or below fixation required either detection or discrimination responses. Cuing effects were determined for the target-elicited P1 component and for the lateralized readiness potential (LRP). Task-dependent cuing effects on reaction time were found in the unimodal but not in the crossmodal version, but not for the P1 component. The LRP data indicated that inhibition of return in the unimodal detection task had a premotoric locus. These findings suggest that inhibition in the unimodal detection task resulted from speeded motor inhibition triggered by the visual cue. PMID- 16712586 TI - An event-related potential study of supramodal attentional control and crossmodal attention effects. AB - We conducted two audiovisual experiments to determine whether event-related potential (ERP) components elicited by attention-directing cues reflect supramodal attentional control. Symbolic visual cues were used to direct attention prior to auditory targets in Experiment 1, and symbolic auditory cues were used to direct attention prior to visual targets in Experiment 2. Different patterns of cue ERPs were found in the two experiments. A frontal negativity called the ADAN was absent in Experiment 2, which indicates that this component does not reflect supramodal attentional control. A posterior positivity called the LDAP was observed in both experiments but was focused more posteriorly over the occipital scalp in Experiment 2. This component appears to reflect multiple processes, including visual processes involved in location marking and target preparation as well as supramodal processes involved in attentional control. PMID- 16712587 TI - Preattentive timing of empty intervals is from marker offset to onset. AB - The internal marker hypothesis explains the superior timing performance for empty intervals over filled intervals by assuming that timing an empty interval starts from the offset of the first marker stimulus and stops at the onset of the second marker stimulus. Other models suggest that timing an empty interval is from first marker onset to second marker onset. We used an electrophysiological measure of preattentive change detection, the mismatch negativity (MMN), to examine processing of empty intervals. Participants watched a silent movie while a stream of auditory stimuli demarcating empty intervals was presented in the background. Most intervals were of the same duration (standards), but shorter intervals (deviants) were presented occasionally also. The pattern of MMN amplitudes obtained across five deviant conditions indicated that preattentive timing of empty intervals occurred from marker offset to onset. PMID- 16712588 TI - Probability-independent and -dependent ERPs reflecting visual change detection. AB - In ERP studies, two posterior components with different polarities have been identified as ERP correlates of visual change detection. To compare these components in terms of sensitivity to the preceding stimulus sequence, two peripheral stimuli of different colors (red and blue) were presented with equal (50:50) or different probabilities (20:80 or 80:20), while 12 participants performed shape discrimination at a central location. A posterior positivity at around 90-140 ms was observed with similar amplitude to all stimuli immediately preceded by a different stimulus. In contrast, a posterior negativity at around 140-180 ms was observed to increase in amplitude with increasing number of preceding different stimuli. These results suggest the existence of probability independent and -dependent change processing in the human visual system. The functional significance is discussed in terms of memory-based comparison and stimulus-specific refractoriness. PMID- 16712589 TI - Sex, outcome expectancy, and cardiovascular response to a masculine challenge. AB - Male and female participants were led to believe they could secure a low or high chance of winning a prize by meeting a modest standard on a purportedly masculine task, that is, a task on which men ostensibly had higher ability. As expected, systolic blood pressure responses measured during performance were greater for women than men when the chance of winning was high, but low for both groups when the chance of winning was low. Similar effects were observed for diastolic and mean arterial pressure responses, although analysis of the mean arterial pressure data produced only a main effect for the chance factor. These results conceptually replicate cardiovascular findings obtained in a previous sex difference study. They also confirm the implication of previous ability perception studies that effort-related cardiovascular responses should be low for both sexes when the importance of meeting a gender-relevant challenge is low. PMID- 16712590 TI - Processing of emotional adjectives: Evidence from startle EMG and ERPs. AB - Affective startle modulation in the electromyographic (EMG), auditory startle evoked potentials, and visually evoked potentials (VEPs) were assessed while subjects evaluated pleasant, unpleasant, and neutral adjectives. Acoustic startle probes were presented at random time points 2.5-4.0 s after word onset. The visual P2 and P3 potentials were generally larger during processing of emotional than of neutral adjectives. In contrast, the late positive component was enhanced and was correlated with larger EMG startle responses and auditory startle evoked potential P3 amplitudes for pleasant words only. During internal cognitive activity, the startle reflex represents a measure of "processing interrupt." Thus the startle tone interrupted processing of particularly pleasant adjectives and caused re-alerting to environmental stimuli. Specific effects for pleasant material may arise from a "positivity offset," favoring responses to pleasant material at lower arousal levels. PMID- 16712591 TI - When does size not matter? Effects of stimulus size on affective modulation. AB - Motivationally relevant stimuli have been shown to receive prioritized processing compared to neutral stimuli at distinct processing stages. This effect has been related to the evolutionary importance of rapidly detecting dangers and potential rewards and has been shown to be modulated by the distance between an organism and a faced stimulus. Similarly, recent studies showed degrees of emotional modulation of autonomic responses and subjective arousal ratings depending on stimulus size. In the present study, affective modulation of pictures presented in different sizes was investigated by measuring event-related potentials during a two-choice categorization task. Results showed significant emotional modulation across all sizes at both earlier and later stages of processing. Moreover, affective modulation of earlier processes was reduced in smaller compared to larger sizes, whereas no changes in affective modulation were observed at later stages. PMID- 16712592 TI - Fear but not awareness predicts enhanced sensory processing in fear conditioning. AB - It is not clear whether enhanced cortical processing of reinforced stimuli as reported by neuroimaging studies is due to expectancy of an aversive event alone or to activation of the fear system. The present study investigated cortical and autonomic responses of aware participants using an instructed fear conditioning design. Steady-state visual evoked fields (ssVEF) and heart rate change were recorded to assess sensory processing and activation of the fear system by reinforced (CS+) and nonreinforced stimuli (CS-). Participants who showed heart rate acceleration demonstrated increased ssVEFs in visual and parietal cortex during CS+ in acquisition trials. Heart rate decelerators did not show enhanced cortical activation with respect to the CS+. Participants in both groups reported awareness of CS-US contingencies. Awareness of stimulus contingency in fear conditioning seems not to be sufficient to elicit enhanced visual cortical processing. PMID- 16712594 TI - First pediatric liver transplantation in Ho Chi Minh City, Vietnam. PMID- 16712595 TI - The ways paved for prophylaxis against de novo hepatitis B virus infection after liver transplantation: still many stones left unturned. PMID- 16712596 TI - Management of infants and young children with combined heart and kidney failure. AB - Most infants and children referred for cardiac transplantation have low cardiac output with concurrent renal hypoperfusion leading to renal insufficiency and failure. This article is a review of the literature of and a single center's experience with combined heart and kidney failure in infants and children less than 10 yr of age. While 39 infants less than 10 yr of age were dialyzed pre- or peri-operatively, none required dialysis support at the time of discharge or in 5 10 yr follow-up. Based on our experience we recommend heart transplant alone in infants and young children with primary heart disease even though they have renal dysfunction. PMID- 16712597 TI - Transition of cognitively delayed adolescent organ transplant recipients to adult care. AB - Transition from pediatric to adult health care is a complicated process that goes far beyond transfer of care. The young person who is intellectually impaired or who must struggle to master daily living skills faces more challenges in this process than other adolescents. Physicians, nurses, social workers, occupational therapists, and other health care providers must work with adolescents, their parents, and teachers to assess abilities and transition needs. Communication skills, capacity for decision making, adherence, vocational and educational issues, sexuality, and relationships should be addressed. Interventions may include the creation of an individualized transition plan, skills training, family counseling, a lengthened transition process, involvement in a support group, health promotion, and teaching of self-care skills. The summary for the adult provider should include information about how best to communicate with the young person and an assessment of capacity to make decisions and self-management skills. PMID- 16712599 TI - Reinnervation after heart transplantation in children: results of short-time heart rate variability testing. AB - AIMS: To detect impairment in short-term heart rate variability (HRV) in children after heart and heart-lung transplantation (TX) as reported in adults. To assess vagal and sympathetic influence on the donor heart rate using frequency domain analysis of HRV. METHODS AND RESULTS: Measurement of short-term HRV was performed in 17 patients (age 16.9+/-3.6, 6.1+/-3.7 yr after TX) and 12 healthy controls (age 14.8+/-3.0 yr). Testing consisted of a resting phase of 15 minutes followed by a tilt phase of 45 min. All HRV parameters were significantly impaired in transplanted patients: RR interval (RRI) 717.2+/-122.5 m/s (controls 827+/-139.7, p<0.05), standard deviation of RR interval (RRI-SD) 20.1+/-15.5 (89.9+/-38.4, p<0.001), RRI at tilt 607.9+/-79.7 (654.0+/-104.7, NS), RRI-SD at tilt 21.1+/ 20.0 (60.4+/-31.4, p<0.001). Low-frequency (LF)/High-frequency (HF) ratio of HRV showed prominent sympathetic influence in TX-patients (3.38+/-5.60 vs. 1.18+/ 0.86, NS) increasing during tilting (5.91+/-8.36 vs. 4.74+/-5.27, NS). In subgroup analysis, 4 yr after TX an increasing sympathetic control of heart rate was observed. CONCLUSION: Short-term HRV is severely impaired in children after TX. If changes are observed, they are time-related and show increasing sympathetic influence starting from 4 yr after TX. PMID- 16712600 TI - Value of soluble adhesion molecules and plasma coagulation markers in assessing transplant coronary artery disease in pediatric heart transplant recipients. AB - BACKGROUND: With an increasing number of heart transplantations (HTx) performed in children and an extended long-term survival of these patients, the importance of transplant coronary artery disease (TCAD) rises in this group of transplant recipients. Reliable serum markers for diagnosis or non-invasive monitoring of this disease in pediatric transplant recipients are still missing. We studied the systemic expression of adhesion molecules as well as plasma coagulation markers and the occurrence of TCAD and/or rejection in pediatric heart transplant recipients. METHODS AND RESULTS: The systemic plasma levels of soluble forms of sVCAM-1 and sICAM-1, d-dimer, tissue factor (TF), prothombin fragments F(1+2), and tissue factor pathway inhibitor (TFPI) were assessed in serial venous blood samples (2-4 per patient) in 50 pediatric transplant recipients children and 63 age- and sex-matched non-transplanted controls. TCAD and rejection were diagnosed angiographically or by combined histological, echocardiographic, or clinical signs, respectively. Plasma levels of sICAM-1 and sVCAM-1, d-dimers and prothrombin fragment F(1+2) but not TF and TFPI were significantly increased in children following HTx compared with non-transplanted controls (p<0.001). Among the transplanted patients, sICAM-1 levels were significantly higher in patients with angiographically detectable TCAD than in patients without evidence of TCAD (p<0.005). Plasma sICAM-1 levels above a cutoff value of 1500 ng/mL (95.5 percentile of control values) were indicative of the presence of TCAD (odds ratio 2.7; 95% confidence interval, 1.34-5.56, p = 0.022; Fisher's exact test). Only d dimers were found to be significantly elevated in children with signs of myocardial rejection compared with those without rejection. CONCLUSIONS: Our results suggest that plasma sICAM-1 and d-dimer levels may be potentially useful to non-invasively assess TCAD and rejection, respectively, in pediatric heart transplant recipients. PMID- 16712601 TI - Pediatric renal transplantation in a South African teaching hospital: a 20-year perspective. AB - INTRODUCTION: Renal transplantation is established as the standard of care for end-stage renal failure (ESRF) in the developed world. In emerging nations, the appropriateness of such costly interventions has been questioned. We undertook an analysis of all renal transplants undertaken under the care of the pediatric nephrology service at the Johannesburg Hospital, South Africa, in order to establish the outcomes of a transplantation service in a resource-constrained environment in a developing country. METHODS: This was a retrospective review of renal transplantation undertaken at a single teaching hospital in Johannesburg, part of the University of the Witwatersrand. Two hundred and eighty-two transplants were performed between 1984 and 2003. Demographic characteristics of the transplanted population, diagnosis, morbidity, graft survival, and mortality were recorded. RESULTS: Overall 1-, 5-, and 10-yr graft survival was 82, 44, and 23%. Overall 1-, 5-, and 10-yr patient survival was 97, 84, and 68%. The median graft survival for all transplantation episodes was 4.38 yr; 70% of patients survive 10 yr and 54% survive 20 yr or more. Although early graft survival was good, there was a more rapid rate of graft loss than when compared to results from developed centers with much poorer results at 5 and 10 yr. Causes of ESRF show marked variation between the races, and black patients have significantly worse outcomes than others. Compared with white patients, black recipients received fewer living donor kidneys (26 vs. 10%, p = 0.0019), a greater proportion of totally mismatched organs (56 vs. 36%, p = 0.015), less pre-emptive transplantation (7 vs. 35%, p = 0.0001) and experienced a higher rate of primary non-function (13 vs. 3%, p = 0.004). Surgical complications of transplantation occurred in 9% of recipients, but rarely led to graft loss. CONCLUSION: Pediatric renal transplantation in Johannesburg can be accomplished with low complication rates, but medium and long-term graft survival is poor when compared with contemporary results achieved in developed countries. The difficulties of undertaking such complex, multidisciplinary interventions in a developing nation are daunting, but we believe that renal transplantation should still be the treatment of choice for all children with ESRF. The poorer outcomes in black recipients can be addressed by increasing education in our communities and expanding the pool of appropriate donors. Better institutional support would allow for improved long-term patient care. PMID- 16712602 TI - C4d in pediatric renal allograft biopsies: a marker for negative outcome in steroid-resistant rejection. AB - Recently, deposition of C4d, reflecting complement activation via the classical pathway, has been established as marker of antibody-mediated rejection. As C4d can be detected in paraffin sections, it allows for retrospective analysis in populations with low case loads, such as in pediatric transplantation. In this study we re-evaluated consecutive renal transplant biopsies obtained since 1990 in 36 children (18 boys, 18 girls) who had received their allograft (nine living, 27 cadaveric) at an age of 10.12+/-4.4 yr. Clinical indications for biopsy were 16 acute steroid resistant rejections (ASRs), 11 chronic rejections and nine other diagnoses. Overall, C4d deposition was found in nine cases (25%), eight of them with diagnosed ASR. Six out of these eight allografts were lost during 36 months of clinical follow-up, a significantly higher rate than in C4d-negative biopsies (p<0.05). C4d status therefore turned out to be an excellent predictor for inferior graft survival following ASR. None of the other histopathologic markers were sensitive for humoral rejections. In conclusion, the high prevalence of C4d-positive staining in ASR demonstrates the importance of the humoral part of the immune system in pediatric transplantation. The worse outcome of C4d positive rejections despite massive immunosuppressive therapy clearly indicates the need for innovative therapies in this high-risk population. PMID- 16712603 TI - Effects of lactational cyclosporine A use on rat pups. AB - We aimed to evaluate the structural and functional changes in the thymus and kidneys of rat pups whose mothers were given cyclosporine A (CsA) during lactational period. Six adult nursing Wistar rats and their 30 pups were studied. Rat pups were divided into four groups as follows: 21-day treated group and 21 day placebo group, each including 10 breastfeeding pups sacrificed on the 21st day, whose mothers were given CsA or placebo, respectively (infancy groups) and, 60-day treated group and 60-day placebo group, each including five breastfeeding pups sacrificed on the 60th day, whose mothers were given CsA or placebo, respectively (puberty groups). While CsA levels of mother rats were very high, CsA levels of 21-day treated group pups were zero. There were no renal histomorphometric differences between study and control pups in both age groups. Renal function parameters showed significant differences between study and control pups in the infancy group: the 21-day treated group pups had significantly lower urine volume, proteinuria, FE(Na) and urinary NAG/creatinine ratio. GFR was also lower in the 21-day treated group, but the difference was not significant, and serum creatinine levels were also not different. Renal function differences were not present among the pubertal pups. Thymic corticomedullary ratio of the 21-day treated group was significantly higher than the 21-day placebo group, while there was no difference between the 60-day treated group and 60-day placebo group. There were no significant changes in the number and distribution of CD3+, CD4+, and CD8+ thymocytes between study and control pups in both age groups. In conclusion, breastfeeding by CsA-treated mother rats induced structural alterations in the thymus and functional changes in the kidneys of the rat pups during infancy. Disturbances in the kidneys and thymus mostly improved after CsA exposure was over. PMID- 16712604 TI - Distinctive characteristics of diabetes mellitus after hematopoietic cell transplantation during childhood. AB - We report on six patients who developed diabetes mellitus after hematopoietic cell transplantation (HCT). The prevalence in our cohort of long-term survivors after HCT performed below 18 yr of age was 3%. The median age at onset of diabetes was 22.4 yr (range 11.3-34.4). The median period between HCT and diabetes was 10.1 yr (range 5.6-22.1). Five out of the six patients received total irradiation therapy and five had other endocrinological abnormalities. The onset of diabetes in all patients was insidious and none had diabetic ketoacidosis. Body mass indexes at diabetes onset were within normal levels. The clinical and laboratory features that characterized our patients with diabetes after HCT make it difficult to classify them as having type-1 or type-2 diabetes. The relatively high prevalence of diabetes and its insidious onset in this group of patients, advocate clinicians to evaluate carefully even slight variations in fasting blood glucose, usually included in the routine biochemistry follow-up. These data also suggest that HbA1c and oral glucose-tolerance test should be added to the follow-up program of late complications if fasting blood glucose levels are slightly increased. PMID- 16712605 TI - Mortality risk score in liver transplantation: changes over time in its predicting power. AB - Since the onset of our liver transplantation program in 1992, 362 transplants were performed in 338 children. A risk score for predicting mortality was designed and implemented over time. The description of a method utilized to design the risk score, changes in mortality rate over 12 yr and the analysis of factors that might have influenced these changes are presented and discussed in this paper. PATIENTS AND METHODS: Cox regression analysis was applied to a retrospective sample of 110 patients with liver cirrhosis, transplanted between 1992 and 2000. A risk score was prepared using beta coefficients of the two significant variables related to survival time: age (1.08, p=0.02) and bilirubin levels (0.93, p=0.03), and two groups were identified: low- and high-risk score. The score was applied in two consecutive samples: 2000-2002 and 2002-2004. RESULTS: In the first sample (1992-2000), we found 69 and 41 as low- and high risk patients, with a median survival time of 93.13 and 2.93 months (p=0.0001). In the 2000-2002 sample, a median survival time of 41.7 and 2.33 months (p=0.03) was found for low- and high-risk groups, respectively. In the third sample (2002 2004), there was a remarkable decrease in mortality in the high-risk group (n=29) and the score did not discriminate between high- and low-risk groups (p=0.35). CONCLUSION: A scoring system to identify risk levels in liver transplantation patients is an operative and powerful tool during a given period of time but it has to be updated as risk factors will vary following the team's learning curve. PMID- 16712606 TI - Conversion from calcineurin inhibitor to sirolimus in pediatric chronic allograft nephropathy. AB - Chronic allograft nephropathy is a major cause for allograft loss in renal transplantation. Sirolimus was recently introduced as a potent non-nephrotoxic alternative to calcineurin inhibitors. In the present study, effects of a conversion protocol were investigated in pediatric chronic allograft nephropathy with declining glomerular filtration rate (GFR), defined by a Schwartz formula clearance below 60 mL/1.73 m(2)/min, steadily increasing serum creatinine and allograft biopsy. In eight children with a median age of 12.8 yr, sirolimus was started at median 32 months after transplantation with a loading dose of 0.24 mg/kg bodyweight (BW), followed by 0.2 mg/kgBW/day, aimed at trough levels of 15 20 ng/mL. Calcineurin inhibitors were reduced to 50% at the start of sirolimus and discontinued at median 7 days when target levels of sirolimus were reached. Following conversion, changes of GFR significantly stabilized (-2.9 vs. +0.4 mL/min/1.73 m(2)/month, p = 0.025). Individual GFR increased in five out of eight patients (p = 0.026), and only one child exhibited unaltered progression of graft failure. In the responders, mean serum creatinine improved by 0.3 mg/dL (p = 0.043). Effects were not dependent on GFR at conversion, or on time post transplantation. Blood pressure, hematological parameters and proteinuria remained stable during the observation period, and serum lipids increased transiently. About half of the children suffered from infectious complications. No child had to be taken off sirolimus; there was no graft loss during the observation period. In conclusion, conversion from calcineurin inhibitors to sirolimus is an effective protocol with tolerable side effects to stabilize renal graft function for at least one yr in the majority of children with biopsy-proven chronic allograft nephropathy. PMID- 16712607 TI - Long-term protection against hepatitis B in pediatric liver recipients can be achieved effectively with vaccination after transplantation. AB - Liver recipients who have antibodies to hepatitis B core antigen (anti-HBc) or received an anti-HBc positive liver graft are at risk of acquiring de novo hepatitis B infection so a life long prophylaxis is required. A post-transplant vaccination against hepatitis B virus (HBV) can offer a better alternative than either hepatitis B hyperimmune globulin (HBIG) or lamivudine. This study investigated the course of anti-HBs titer after vaccination and analyzed the factors that influence the response. Between October 1999 and February 2003, 37 pediatric patients were given a post-transplant vaccination when an anti-HBc positive graft was used, the recipient was anti-HBc positive, or when anti-HBs titer was below 20 IU/L irrespective of the serological status. Thirty-three patients responded to the vaccine and did not require further HBIG injections at a mean follow up of 33.6 months. Fifteen patients were good responder and only needed a single set of vaccines, and 18 were poor responder needing additional boosters. Two patients developed de novo hepatitis B infection and two required additional HBIG injections. Preoperative severity of liver disease, serological status of HBV of recipient or donor, use of HBIG or pulse steroid therapy, type of vaccines, and dose or time interval between doses had no influence on response rate. Recipients with a high preoperative anti-HB titer, small graft-recipient weight ratio (GRWR), greater catch up growth, heavier body weight, lower tacrolimus level at the time of vaccination, and longer time interval between transplant or steroid medication and vaccination yielded good response. If well tailored, post-transplant vaccination can be effective and offer patients prophylaxis against de novo hepatitis B infection for a prolonged period of time. PMID- 16712608 TI - Children presenting with end-stage renal disease of unexplained etiology: implications for disease recurrence after transplantation. AB - With longer graft and patient survival, recurrent disease is becoming recognized as an increasingly important contributor to long-term graft loss in renal transplant recipients. However, patients may present for the first time in end stage renal disease (ESRD) leading to uncertainty as to their underlying diagnosis and the risk of recurrence. The purpose of this study was to describe the features of children who presented for the first time in ESRD and to determine the predictive value of investigations in differentiating diseases with and without a recurrence risk. From 7/99 to 11/04, 13 children presented to our center in ESRD. Their median age was 13.3 yr; 77% were male. The majority were hypertensive (77%) and oligoanuric (69%). All had proteinuria (median urine protein to creatinine ratio [Up/c] 7.0), and 92% had microhematuria. Only seven had small kidneys on ultrasound. All children underwent a serologic work-up and six (46%) were biopsied. Of the 13 children, seven had a glomerular disease; in five the diagnosis was made on biopsy, in one on serologic testing and one by family history. Of the remaining six children, three had non-glomerular diseases: obstructive uropathy in one and primary hyperoxaluria type 1 in two, and 3 had an unknown disease. When patients with glomerular diseases were compared with those with non-glomerular diseases, the two predictors for glomerular disease were a lower serum albumin (p = 0.004) and a higher serum bicarbonate level (p = 0.01). Comparing patients with and without a risk of recurrence, there were no differences between the two groups in any of their demographic, clinical, or biochemical parameters by analysis of variance (including serum albumin or proteinuria). In summary, the vast majority of children presenting in ESRD have hematuria and proteinuria, even with non-glomerular diseases. The significant overlap in clinical features between patients with and without a risk of recurrence emphasizes the need for all children presenting in ESRD to be evaluated extensively so that disease recurrence after transplantation can be anticipated or even prevented. PMID- 16712609 TI - Quality of life following allogeneic stem cell transplantation, comparing parents' and children's perspective. AB - There is insufficient knowledge about the quality of life (QoL) among children after allogeneic stem cell transplantation (SCT). We recently reported an overall, good self-assessed QoL and health in 52 children who were three yr or more beyond SCT. The focus of this paper is the QoL as assessed by their parents, of whom 42 participated in the study. Using Swedish child health questionnaire (SCHQ)-PF50, parents rated their children's QoL lower on both the psychosocial (p<0.001) and physical summary scales (p<0.001) than the normative group of parents of children without chronic disease. Although essentially following each other, parent scores tended to be lower than children's own SCHQ-CF87 scores, particularly in the domains 'role socially due to physical limitations' (p<0.01) and 'self-esteem' (p<0.05). In the 'bodily pain' domain, patients' and parents' low scores agreed. The child's condition had a greater impact on parents' emotional situation than in the norm population (p<0.001). The severity of the child's physician-rated late effects (p<0.05) or of self-assessed subjective symptoms (p<0.01-0.05) was associated with a lower parental rating of the child's QoL. High Lansky or Karnofsky scores corresponded, respectively, to higher psychosocial (p<0.05) and physical (p<0.05) summary scores. It is concluded that as children, parents, and clinicians do not necessarily adopt similar views of a child's illness and of its impact on the child's life, clarity with regard to who is responsible for assessing the child's QoL is crucial when interpreting pediatric QoL studies. PMID- 16712610 TI - Unrelated donor cord blood transplantation for childhood severe aplastic anemia after a modified conditioning. AB - Treatment of severe aplastic anemia (SAA) patients who lack human leukocyte antigen (HLA)-matched donors and failed immunosuppressive therapy (IST) is challenging. Recently, umbilical cord blood transplantation (CBT) after non myeloablative therapy has been reported in adult but not in childhood SAA. However, most cases resulted in mixed donor chimerism and incomplete hematological recovery. We reported an 11-yr-old girl with recurred SAA 5 yr after IST who underwent unrelated donor CBT after a modified regimen. This patient had renal and cardiac dysfunction, and lacked suitable bone marrow donors. The 3.9 x 10(7)/kg CB cells from an HLA one-locus mismatched unrelated donor were infused after conditioning with total body irradiation (5 Gy), melphalan (120 mg/m(2)), and fludarabin (120 mg/m(2)). Hematological recovery was favorable in complete chimerism. A major complication was only skin graft-versus host disease (grade I). CB could be an alternate stem cell source for childhood SAA after modified preparative regimen. PMID- 16712611 TI - Successful unrelated mismatched cord blood transplantation in an infant with severe combined immunodeficiency and Mycobacterium bovis bacillus Calmette-Guerin disease. AB - The case reported here of an infant who presented with Pneumocystis carinii pneumonia, CD4+ lymphopenia, and hypogammaglobulinemia attributable to severe combined immunodeficiency (SCID). This report discussed treatment of Mycobacterium bovis bacillus Calmette-Guerin disease with unrelated cord blood transplantation in addition to antituberculous therapy, by adoptively transferring donor immunity with induction of mixed chimerism. Because of the unique nature of umbilical cord blood hematopoietic cells, engraftment without conditioning may be possible in SCID patients without fully matched donors. PMID- 16712612 TI - Central nervous system lymphoproliferative disorder in pediatric kidney transplant recipients. AB - Post-transplant lymphoproliferative disorder (PTLD) is a complication of transplantation resulting from impaired immune surveillance because of pharmacologic immunosuppression. We present two cases of central nervous system (CNS) PTLD in children on calcineurin-inhibitor free immunosuppression with dramatically different presentations and outcomes. One patient had brain and spinal cord lymphoma with a rapid and fatal course. The other patient had brain and ocular PTLD that responded to multimodal therapy with reduction of immunosuppression, high-dose steroids, and rituximab given in a dose-escalation protocol. This protocol may have enhanced the penetration of rituximab into the CNS. We review the literature on CNS and ocular PTLD and elaborate on the treatments available for both diseases. PMID- 16712613 TI - Liver transplant in a four-month-old child with biliary atresia, unilateral pulmonary agenesis, and diaphragmatic hernia: first case report. AB - Bilateral pulmonary agenesis (PA) is a rare embryological defect incompatible with life. Unilateral PA has a wide range of clinical presentations: its prognosis depends on the presence and severity of other associated anomalies. Fetal biliary atresia has been associated with a number of congenital anomalies, but the etiology is still not understood. An unusual case of a child with right PA, right diaphragmatic hernia, and delayed diagnosed biliary atresia leading to liver failure is presented herein. At the age of 4 months the patient was referred to the Transplant Department at Children Healthcare of Atlanta at Egleston with cholestasis and failure to thrive. With a rapidly progressive liver insufficiency, this child was evaluated for liver transplantation. In the absence of any respiratory symptom, the patient received a deceased donor size-matched left lateral segment liver transplant, which covered the diaphragmatic defect, with no further repair required. Twenty-seven months post-transplant, the patient has good graft function, a normal Z-score and is thriving. In spite of the increased physiological and surgical challenges (absence of right lung tissue, hemi-diaphragm, and ectopic position of the liver in the right chest), liver transplantation was performed with positive outcome in this high-risk child. Whether PA, may have developmentally contributed to expression of biliary atresia will need further investigation. PMID- 16712614 TI - The efficacy of liver transplantation in malignant liver tumors associated with tyrosinemia: clinical and laboratory findings of five cases. AB - To evaluate clinical and laboratory findings of these patients and the efficacy of liver transplantation in children with hepatocellular carcinoma (HCC) and hepatoblastoma (HB) associated with tyrosinemia. Among 113 children with liver tumors diagnosed between 1972 and 2004 five patients had HCC or HB associated with tyrosinemia. The age at diagnosis of the HCC or HB ranged from 9.5 to 17 yr and male:female ratio was 1:4. During regular clinic visits for tyrosinemia, elevated alpha-fetoprotein (AFP) was detected in all patients. AFP levels ranged between 13.7 and 29 340 IU/mL. Radiological studies including ultrasound, computed tomography and magnetic resonance imaging showed heterogeneous parenchyma and nodules in the liver. The patients did not have any metastatic disease. The time from diagnosis of tyrosinemia to HCC or HB ranged from 9.25 to 15.25 yr. Histopathologically, four patients have been diagnosed as HCC and one patient had HB. All patients were given chemotherapy including cisplatin and adriamycin. In three patients, living-related liver transplantation was performed. They had no treatment after transplantation. All of them are disease free. One patient was treated with chemotherapy and right hepatectomy. She had no suitable donor for living-related liver transplantation. Three months after completing chemotherapy, she had recurrent tumor in the left lobe of the liver and she died with progressive disease. The last patient whose parents were not suitable as donors for living-related liver transplantation is waiting for a deceased donor graft. All patients had limited disease to liver due to close clinical and radiological follow up for tyrosinemia. In these patients liver transplantation is curative both for liver tumor and tyrosinemia. PMID- 16712615 TI - Living-related liver transplantation with removal of inferior vena cava for unresectable hepatoblastoma. AB - We report a case of a two-yr-old boy with hepatoblastoma resectable only by total hepatectomy including the vena cava. Successful LTx was performed with a living donor segment without vena cava reconstruction. The tumor was located in the bilateral lobe, surrounding the IVC. In spite of the high-dose chemotherapy, the tumor did not become resectable. LTx was performed using left lateral segment after removal of the IVC combined with total hepatectomy. Because the collaterals were well developed, the patient tolerated the procedure well. The serum AFP level decreased from 186 699 to 8 ng/mL in 11 months after LTx without local recurrence or distant metastasis. PMID- 16712616 TI - Propylthiouracil-associated liver failure presenting as probable autoimmune hepatitis in a child with Graves' disease. AB - This case describes a young girl with Graves' disease, who presented with fulminant hepatic failure 9 months into propylthiouracil (PTU) therapy. Her clinical presentation was consistent with 'probable autoimmune hepatitis,' as defined by the International Autoimmune Hepatitis Group scoring system. Despite discontinuation of PTU and high-dose steroid therapy, she required liver transplantation. Subsequent pathology could not definitively rule out autoimmune hepatitis. PTU is an important cause of drug-related liver failure in children, and clinicians should be mindful that it is frequently used in patients who already have an underlying risk of autoimmune liver disease. PMID- 16712617 TI - Food protein sensitivity with partial villous atrophy after pediatric liver transplantation with tacrolimus immunosuppression. AB - We report three pediatric liver transplant recipients receiving tacrolimus immunosuppression presented with vomiting, heme-positive stools and failure to thrive, who had subtotal villous atrophy in their histology because of food protein sensitivity. Case findings and current literature of the casual relationship between tacrolimus and food allergies briefly reviewed. PMID- 16712619 TI - Empathy: a timeless skill for the pain medicine toolbox. PMID- 16712621 TI - The association between psychopathology and placebo analgesia in patients with discogenic low back pain. AB - BACKGROUND: Chronic low back pain patients have a high rate of psychopathology, comprised mainly of depression, anxiety, and high levels of neuroticism. We previously found that psychopathology is associated with increased placebo analgesia in this patient group. OBJECTIVE: To better understand this finding in the context of other known predictors of placebo response (such as expectations for relief), we performed a detailed analysis of expectations and other possible covariates. DESIGN: We conducted a double-blind, placebo-controlled, randomized, crossover-designed trial of intravenous morphine vs. placebo in 60 subjects with chronic low back pain and discogenic abnormalities. Patients were stratified into three groups of psychiatric symptom severity (Low, Moderate, and High), based on composite scores on depression, anxiety for pain, and neuroticism scales. Subjects were given intravenous morphine and placebo in random order on separate visits, and completed serial pain ratings over 3 h at each session. RESULTS: With 20 subjects per group, there were small, but significant differences between groups in baseline pain ratings. No differences were found between groups in age, gender, and radicular pain. Patients with low psychological symptomatology reported 7.7% total pain relief with placebo compared with 23.4% in the Moderate group and 23.5% in the High group (P < 0.05). Expectations were not significantly different between groups, but in the High group expectation levels predicted placebo analgesia (P < 0.001). Neuropathic pain quality was also a predictor (P < 0.05). CONCLUSIONS: This study indicates that high and moderate levels of psychopathology are associated with heightened placebo analgesia in chronic low back pain patients. Expectations were only an influence in the high psychopathology group, and neuropathic pain affects placebo responses. These findings have implications for future research characterizing placebo responders. PMID- 16712622 TI - Cluster analysis of SF-36 scales as a predictor of spinal pain patients response to a multidisciplinary pain management approach beginning with epidural steroid injection. AB - OBJECTIVE: To investigate whether grouping of patients with back pain into similar behavioral patient profiles using SF-36 scores is predictive of outcome following 1-year treatment in a multidisciplinary spine center beginning with referral for epidural steroid injection. DESIGN: A prospective observational study was conducted on 81 consecutive patients selected for epidural steroid injections by independent physicians following common institutional criteria. Each patient completed a baseline SF-36 questionnaire as well as a numerical response pain scale. The initial SF-36 data were used to place each patient into one of three subgroups (Highly Functional, Emotional Adapters, and Dysfunctional). Follow-up SF-36 and numerical response pain scale questionnaires were completed by the patients at 1 month and 12 months following the initial epidural steroid injection. RESULTS: Results revealed significant improvement among all three patient subgroups following multidisciplinary treatment at both 1 month and 12 months. Few differences in outcome occurred among the subgroups. CONCLUSION: The SF-36-determined subgroups did not predict response to a multidisciplinary pain clinic. All three subgroups showed similar improvement following treatment. PMID- 16712624 TI - Postherpetic pain: more than sensory neuralgia? AB - OBJECTIVE: To describe a series of older adult patients with postherpetic myofascial pain, a heretofore rarely described complication of herpes zoster. DESIGN: Case series. SETTING: Outpatient older adult pain clinic. PATIENTS: Five older adults are presented with myofascial pain that developed as a complication of herpes zoster. RESULTS: Pain duration at the time of presentation ranged from 4 months to 7 years. All patients reported functional impairment from pain despite oral analgesics. Myofascial pathology was diagnosed by the presence of taut bands and trigger points in the affected myotome. Upon successful treatment of the myofascial pain with nonpharmacologic modalities (e.g., physical therapy, trigger point injections, dry needling, and/or percutaneous electrical nerve stimulation), all patients reported symptomatic improvement, and four out of five were able to significantly reduce or discontinue their opioids. CONCLUSION: Postherpetic pain is traditionally conceptualized as a purely sensory phenomenon. Identification of the intrusion of a myofascial component may be worthwhile, both from the standpoint of enhanced pain relief and reduction in the need for oral analgesics. Formal exploration of this phenomenon is needed. PMID- 16712623 TI - Comparative effectiveness of cervical transforaminal injections with particulate and nonparticulate corticosteroid preparations for cervical radicular pain. AB - OBJECTIVES: Cervical transforaminal epidural injections of corticosteroids have been used in the treatment of radicular pain. Particulate agents have been associated with rare adverse neurological outcomes. It is unknown whether nonparticulate preparations are any less effective than particulate preparations. Therefore, a study was designed to determine whether there is a basis for promoting a theoretically safer nonparticulate corticosteroid preparation. DESIGN: Volunteer patients were randomized to receive a single cervical transforaminal epidural injection with one of two corticosteroid preparations. SETTING: This study was undertaken in a private practice setting. PATIENTS: Those with single-level, unilateral radicular pain with advanced imaging demonstrating single-level neural compression. INTERVENTIONS: Patients received a single cervical transforaminal epidural injection with either dexamethasone or triamcinolone. OUTCOME MEASURES: Ratings were obtained by an independent unbiased assessor at 4 weeks via a telephone interview. A visual analog pain scale was used preprocedurally and a verbal integer scale was used at 4 weeks to assess the severity of the patient's radicular pain. As a secondary outcome measure, a patient-specified functional outcome measure was obtained. RESULTS: Both groups exhibited statistically and clinically significant improvements in pain at 4 weeks. Although the triamcinolone group exhibited a somewhat greater improvement, the difference between groups was not significantly different. CONCLUSION: The study found that the effectiveness of dexamethasone was slightly less than that of triamcinolone, but the difference was neither statistically nor clinically significant. A theoretically safer nonparticulant agent appears to be a valid alternative to particulate agents that have been used to date, and which have been associated with hazard. PMID- 16712626 TI - A review of intrathecal fentanyl and sufentanil for the treatment of chronic pain. AB - Intrathecal infusion of morphine using implantable pumps is an accepted practice for long-term management of chronic pain. Despite clinical benefit, development of tolerance and side-effects associated with intrathecal morphine has prompted investigators to explore alternative opioids such as the potent anilinopiperidine analogs, fentanyl, and sufentanil. Relevant preclinical and clinical literature from the MEDLINE database was used primarily for this review. In vitro, both compounds are stable in solution, but studies have not been conducted using implantable pumps under simulated use conditions (e.g., long-term stability at body temperature). Preclinical studies of limited duration have demonstrated efficacy, but safety-toxicology studies have been limited to intermittent boluses of sufentanil only. Few clinical reports on the use of intrathecal sufentanil or fentanyl for chronic pain are available. Although results confirm potency and efficacy with intrathecal administration, further studies are needed to support the long-term use of either opioid in chronic pain management. PMID- 16712627 TI - Treatment of refractory, chronic low back pain with botulinum neurotoxin A: an open-label, pilot study. AB - OBJECTIVE: To study the short- and long-term effects of botulinum neurotoxin A (BoNT-A, Botox, Allergan Inc.) on refractory chronic low back pain. DESIGN: The effect of botulinum neurotoxin A on chronic low back pain was prospectively studied in 75 patients with repeated treatments over a period of 14 months. Pain intensity (visual analog scale [VAS]), pain frequency (pain days), and perceived functional status (Oswestry scale) were assessed at baseline, 3 weeks, and at 2, 4, 6, 8, 10, 12, and 14 months. BoNT-A was injected into para-spinal muscles at 4 5 levels (between L1 and S1) unilaterally or bilaterally. The dose per site varied from 40 to 50 units. The total dose per session ranged from 200 to 500 units. Reinjections were performed at 4 months only when pain returned. RESULTS: At 3 weeks, 40 patients (53%) and at 2 months, 39 patients (52%) reported significant pain relief. The change in VAS, Oswestry score, and pain days was significant compared with baseline at 2 months after each injection period (P < 0.005) and remained so over subsequent treatments. Among initial responders, 91% continued responsiveness over the length of the study. Three patients (4%), after the first treatment, had a mild flulike reaction that lasted 2-5 days. CONCLUSION: Botulinum neurotoxin A may be beneficial in patients with chronic low back pain. A favorable initial response predicts subsequent responsiveness. The treatment is well tolerated, and side effects are mild and transient. PMID- 16712628 TI - Empathy in the physician's pain practice: benefits, barriers, and recommendations. PMID- 16712630 TI - Annotation: Tourette syndrome: a relentless drumbeat--driven by misguided brain oscillations. AB - OBJECTIVE: This annotation reviews recent evidence that points to the likely role of aberrant neural oscillations in the pathogenesis of Tourette syndrome (TS). METHODS: The available anatomic and electrophysiological findings in TS are reviewed in the context of an emerging picture of the crucial role that neural oscillations play in maintaining normal central nervous system (CNS) function. RESULTS: Neurons form behavior-dependent oscillating networks of various sizes and frequencies that bias input selection and facilitate synaptic plasticity, mechanisms that cooperatively support temporal representation as well as the transfer and long-term consolidation of information. Coherent network activity is likely to modulate sensorimotor gating as well as focused motor actions. When these networks are dysrhythmic, there may be a loss of control of sensory information and motor action. The known electrophysiological effects of medications and surgical interventions used to treat TS likely have an ameliorative effect on these aberrant oscillations. Similarly, a strong case can be made that successful behavioral treatments involve the willful training regions of the prefrontal cortex to engage in tic suppression and the performance of competing motor responses to unwanted sensory urges such that these prefrontal regions become effective modulators of aberrant thalamocortical rhythms. CONCLUSIONS: A deeper understanding of neural oscillations may illuminate the complex, challenging, enigmatic, internal world that is TS. PMID- 16712631 TI - Fine-motor skill deficits in childhood predict adulthood tic severity and global psychosocial functioning in Tourette's syndrome. AB - BACKGROUND: Most children with Tourette's syndrome (TS) experience a significant decline in tic symptoms during adolescence. Currently no clinical measures have been identified that can predict whose tic symptoms will persist into adulthood. Patients with TS have deficits on neuropsychological tests involving fine-motor coordination and visual-motor integration. We seek to determine if these neuropsychological tests are useful in predicting future symptom severity. METHODS: Thirty-two children, aged 8-14, with TS underwent clinical evaluation and a focused neuropsychological testing battery consisting of the Purdue Pegboard, Beery Visual-Motor Integration (VMI) Test and the Rey-Osterreith Complex Figure Task (RCFT). A follow-up clinical assessment was performed on these children an average of 7.5 years later. Ordinal logistic regression analysis was used to correlate neuropsychological testing at Time 1 with tic severity, OCD severity and global psychosocial functioning at Time 2. RESULTS: Poor performance with the dominant hand on the Purdue Pegboard test predicted worse adulthood tic severity and correlated with tic severity at the time of childhood assessment. Poor performance on the VMI and Purdue Pegboard tests (both dominant and non-dominant hand) also predicted worse adulthood global psychosocial functioning. None of the neuropsychological tests were useful in predicting the future course of OCD symptoms in TS patients. CONCLUSION: Fine motor skill deficits may be a predictor of future tic severity and global psychosocial function in children with TS. We hypothesize that performance on the Purdue Pegboard test may serve as a useful endophenotype in the study of TS and provide a rough measure of the degree of basal ganglia dysfunction present in TS patients. PMID- 16712632 TI - Event rate and event-related potentials in ADHD. AB - BACKGROUND: It has been repeatedly found that performance of children with attention deficit hyperactivity disorder (ADHD) is more impaired when a long inter-stimulus interval (ISI) is used than when a short ISI is used. According to the cognitive-energetic model, this may reflect difficulty in remaining in an optimal motor activation state because of insufficient effort allocation. METHOD: Event-related potentials (ERPs) were evaluated during a Go/No-Go task that incorporates a condition with a fast and a slow presentation rate. RESULTS: ADHD, whether or not comorbid with oppositional defiant/conduct disorder (ODD/CD), was associated with a steeper increase in reaction time (RT) from the fast to the slow condition accompanied by a missing increment of the parietal P3. Speed of responding was found to be correlated with P3 amplitude. In the fast condition, children with ADHD made more errors of commission, accompanied by a smaller No-Go N2, a component thought to be related to inhibition; however, after controlling for ODD/CD these differences disappeared. CONCLUSIONS: The association between the steeper increase in RT and reduced parietal P3s may indicate that the children with ADHD did not allocate enough extra effort to adjust to a potentially under-activated state. However, the event rate effects could not account for all of the differences between groups and also early automatic information processing stages seem disturbed in this disorder as indexed by larger P2 amplitudes. Alternative explanations are discussed. PMID- 16712633 TI - Colour perception in ADHD. AB - Attention-deficit/hyperactivity disorder (ADHD) is associated with unexplained impairments on speeded naming of coloured stimuli. These deficits may reflect hypofunctioning retinal dopaminergic mechanisms impairing particularly blue yellow colour discrimination. Colour perception and rapid colour naming ability were investigated in 14 children with ADHD and 13 healthy peers matched for age, gender, and IQ, using the Farnsworth-Munsell 100 Hue Test (FMT) and the Stroop Colour-Word test. Children with ADHD committed more errors on the FMT, particularly on discrimination of colours along the blue-yellow axis, and were slower on Stroop subtests involving colour naming. However, the latter deficit was accounted for similarly by blue-yellow and red-green discrimination abilities. Blue-yellow colour perception problems in ADHD contribute to but do not fully explain the observed slowed colour naming. PMID- 16712634 TI - Repetitive and stereotyped behaviours in pervasive developmental disorders. AB - BACKGROUND: Repetitive and stereotyped behaviours are a heterogeneous group of behaviours present in many neuropsychiatric disorders. Despite their core significance in PDD, it is not clear whether there are distinct groups of these behaviours with different specificity to autism. METHODS: A two-factor model of the repetitive behaviours, namely sensory/motor (lower-order) and cognitive rigidity (higher-order), was conceptualised. The model's goodness of fit to the data was examined in a clinic population of children with PDD, with and without mental retardation, and of those with only mental retardation (MR). RESULTS: Subjects with PDD had higher rates of all repetitive behaviours compared to those with MR only. The existence of two independent 'lower-order' and 'higher-order' sub-groups of the repetitive behaviours was confirmed only in the MR group. The lower-order behaviours appear to be associated more with global developmental problems, whereas the higher-order behaviours were significantly associated with ruminations in the PDD group. CONCLUSIONS: This study suggests that there may be two distinct sub-groups of repetitive behaviours whereby the sensory and motor repetitive behaviours are generally associated with lower developmental age and less specific to the autistic syndrome whereas the 'higher-order' behaviours may be a more 'autism specific' feature. The co-occurrence of the lower- and higher order behaviours in PDD might reflect the end result of diffuse altered neural networks in these disorders thus being a specific feature of PDD. There is a need for more systematic studies of these behaviours not only in autistic disorders but also in other neuropsychiatric disorders. PMID- 16712635 TI - Investigating the structure of the restricted, repetitive behaviours and interests domain of autism. AB - BACKGROUND: The Restricted, Repetitive Behaviours and Interests (RRBIs) are represented in the DSM-IV and measured by the Autism Diagnostic Interview-Revised (ADI-R) as one of the three homogeneous symptom categories of Pervasive Developmental Disorders. Although this conceptualisation is well accepted in the field, the grouping of symptoms is based primarily on clinical judgment rather than on empirical evidence. METHODS: The objective of this study was to examine the factor structure of the RRBI domain of autism. Eleven items from this domain of the ADI-R were used in a Principal Components Analysis (PCA). Our sample consisted of 339 individuals with a Best Estimate diagnosis of Pervasive Developmental Disorder (PDD). RESULTS: Findings indicate that the RRBI domain is composed of two distinct factors or dimensions: Insistence on Sameness (IS) and Repetitive Sensory and Motor Behaviours (RSMB). RSMB is negatively correlated with adaptive skills; that is, lower functioning individuals tend to have higher levels of repetitive sensory and motor behaviours. On the other hand, IS is positively correlated with autistic symptoms in the communication and language domain. Further analyses suggest moderate familial aggregation among affected sibling pairs within the IS but not the RSMB factor. CONCLUSIONS: These results provide evidence for the heterogeneity of the RRBI domain of the ADI-R in terms of both clinical presentation and other correlates. In addition, the IS factor seems to be under familial (presumably genetic) control, while RSMB appears to simply reflect variation in developmental level. PMID- 16712636 TI - Sensory Experiences Questionnaire: discriminating sensory features in young children with autism, developmental delays, and typical development. AB - BACKGROUND: This study describes a new caregiver-report assessment, the Sensory Experiences Questionnaire (SEQ), and explicates the nature of sensory patterns of hyper- and hyporesponsiveness, their prevalence, and developmental correlates in autism relative to comparison groups. METHOD: Caregivers of 258 children in five diagnostic groups (Autism, PDD, DD/MR, Other DD, Typical) ages 5-80 months completed the SEQ. RESULTS: The SEQ's internal consistency was alpha' = .80. Prevalence of overall sensory symptoms for the Autism group was 69%. Sensory symptoms were inversely related to mental age. The Autism group had significantly higher symptoms than either the Typical or DD groups and presented with a unique pattern of response to sensory stimuli -hyporesponsiveness in both social and nonsocial contexts. A pattern of hyperresponsiveness was similar in the Autism and DD groups, but significantly greater in both clinical groups than in the Typical group. CONCLUSION: The SEQ was able to characterize sensory features in young children with autism, and differentiate their sensory patterns from comparison groups. These unique sensory patterns have etiological implications, as well as relevance for assessment and intervention practices. PMID- 16712638 TI - Joint attention and symbolic play in young children with autism: a randomized controlled intervention study. AB - BACKGROUND: Delays and deficits in joint attention and symbolic play constitute two important developmental problems in young children with autism. These areas of deficit have been well studied in autism but have rarely been the focus of treatment efforts (see Kasari, Freeman, & Paparella, 2001). In this study, we examine the efficacy of targeted interventions of joint attention and symbolic play. METHODS: Participants were 58 children with autism aged 3 and 4 years (46 boys). Children were randomized to a joint attention intervention, a symbolic play intervention, or control group. Interventions were conducted 30 minutes daily for 5-6 weeks. Both structured assessments of joint attention and play skills and mother-child interactions were collected pre and post intervention by independent assessors. RESULTS: Results indicate that both intervention groups improved significantly over the control group on certain behaviors. Children in the joint attention intervention initiated significantly more showing and responsiveness to joint attention on the structured joint attention assessment and more child-initiated joint attention in the mother-child interaction. The children in the play group showed more diverse types of symbolic play in interaction with their mothers and higher play levels on both the play assessment and in interaction with their mothers. CONCLUSIONS: This randomized controlled trial provides promising data on the specificity and generalizability of joint attention and play interventions for young children with autism. Future studies need to examine the long-term effects of these early interventions on children's development. PMID- 16712637 TI - Social behavior and cortisol reactivity in children with fragile X syndrome. AB - OBJECTIVE: To examine the association between limbic-hypothalamic-pituitary adrenal (L-HPA) axis reactivity and social behavior in children with fragile X syndrome (FXS). METHOD: Salivary cortisol changes and concurrent anxiety-related behaviors consistent with the behavioral phenotype of FXS were measured in 90 children with the fragile X full mutation and their 90 unaffected siblings during a social challenge task in the home. RESULTS: Boys and girls with FXS demonstrated more gaze aversion, task avoidance, behavioral signs of distress, and poorer vocal quality than the unaffected siblings. Multiple regression analyses showed that after accounting for effects of IQ, gender, age, quality of the home environment, and basal cortisol level, cortisol reactivity to the task was significantly associated with social gaze in children with FXS. The most gaze aversive children with FXS had cortisol reductions, whereas those with more eye contact demonstrated the most cortisol reactivity. Unaffected siblings demonstrated an opposite pattern in which less eye contact was associated with increased cortisol reactivity. CONCLUSIONS: Results of the study suggest a unique relation between abnormal gaze behavior and L-HPA mediated stress reactivity in FXS. PMID- 16712639 TI - The prevalence of autistic spectrum disorders in adolescents with a history of specific language impairment (SLI). AB - BACKGROUND: Traditionally, autism and specific language impairment (SLI) have been regarded as distinct disorders but, more recently, evidence has been put forward for a closer link between them: a common set of language problems, in particular receptive language difficulties and the existence of intermediate cases including pragmatic language impairment. The present study aimed to examine the prevalence of autism spectrum disorders in a large sample of adolescents with a history of SLI. METHOD: The presence of autism spectrum disorders was examined in seventy-six 14-year-olds with a confirmed history of SLI. A variety of instruments were employed, including the Autism Diagnostic Interview-Revised (ADI R), the Autism Diagnostic Observation Schedule (ADOS) and the Family History Interview (FHI). RESULTS: The prevalence of autism spectrum disorders in young people with SLI was found to be 3.9%, about 10 times what would be expected from the general population. In addition, a much larger number of young people with a history of SLI showed only some autism spectrum symptoms or showed them in a mild form. CONCLUSIONS: Young people with SLI have an increased risk of autism. The magnitude of this risk is considerable. In addition, a larger proportion (a quarter of individuals) present with a number of behaviours consistent with autism spectrum disorders. PMID- 16712640 TI - Development in infants with autism spectrum disorders: a prospective study. AB - BACKGROUND: Autism is rarely diagnosed before three years of age despite evidence suggesting prenatal abnormalities in neurobiological processes. Little is known about when or how development becomes disrupted in the first two years of life in autism. Such information is needed to facilitate early detection and early intervention. METHODS: This prospective study of autism spectrum disorders (ASD) examined development using the Mullen Scales of Early Learning (MSEL) in 87 infants tested at target ages 6, 14, and 24 months. Participants came from infants at high risk (siblings of children with autism) and low risk (no family history of autism) groups. Based on language test scores, Autism Diagnostic Observation Schedule, and clinical judgment at 24 months of age, participants were categorized as: unaffected, ASD, or language delayed (LD). Longitudinal linear regression and ANOVA models were applied to MSEL raw scores, and estimates were compared between the three diagnostic groups. RESULTS: No statistically significant group differences were detected at 6 months. By 14 months of age, the ASD group performed significantly worse than the unaffected group on all scales except Visual Reception. By 24 months of age, the ASD group performed significantly worse than the unaffected group in all domains, and worse than the language delayed group in Gross Motor, Fine Motor, and Receptive Language. The developmental trajectory of the ASD group was slower than the other groups', and showed a significant decrease in development between the first and second birthdays. CONCLUSIONS: Variations from typical and language delayed development are detectable in many children with ASD using a measure of general development by 24 months of age. Unusual slowing in performance occurred between 14 and 24 months of age in ASD. PMID- 16712641 TI - The broader cognitive phenotype of autism in parents: how specific is the tendency for local processing and executive dysfunction? AB - BACKGROUND: The objective of this study was to investigate the tendency for local processing style ('weak central coherence') and executive dysfunction in parents of subjects with an autism spectrum disorder (ASD) compared with parents of individuals with early onset schizophrenia (EOS) and mental retardation (MR). METHOD: Sixty-two parents of subjects with ASD, 36 parents of subjects with EOS and 30 parents of subjects with MR were examined. Data on two scales indicative of local visual processing (Embedded Figures Test, Block Design) and on three executive function tests (Wisconsin Card Sorting Test, Tower of Hanoi, Trailmaking Test) were collected for all participants. RESULTS: Parents of subjects with ASD performed significantly faster on the Embedded Figures Test compared with both control samples. No other substantial group differences were observed. CONCLUSIONS: The findings indicate that an increased tendency for local processing in terms of visual disembedding could be a relatively specific core feature of the broader cognitive phenotype of autism in parents. PMID- 16712642 TI - Pointing the way. PMID- 16712643 TI - Biobanks and registries for HSCT research: potential for future individualized medicine. PMID- 16712645 TI - ELISA methods detect HLA antibodies with variable sensitivity. AB - The accuracy of human leukocyte antigen (HLA) antibody screening results is dependent on the technique employed. In this study, a total of 612 serum samples from patients awaiting kidney transplantation were tested by two different enzyme linked immunosorbent assay (ELISA) methods: LAT-M(R) (One Lambda/BMT) and AbScreen(R) (Biotest). The results of these assays were identical for HLA class I and II antibodies in 524 cases (85.6%), and discrepant in the remaining 88 cases (42 class I, 24 class II, 22 class I and II). More specifically, class I results were in agreement in 26 positive and 522 negative cases and in disagreement in 64 cases. Class II results were the same in 50 positive and 516 negative cases and different in 46 cases. Retesting the samples using LAT 1288 (One Lambda/BMT) and considering previous HLA antibody test results and the history of immunising events, the sensitivity and specificity, respectively, of the two methods were determined as follows: class I: 89.6% and 97.5% (LAT-M) vs. 70.6% and 96.8% (AbScreen), class II: 96.5% and 98.4% (LAT-M) vs. 88.3% and 95.5% (AbScreen). These results indicate that a single ELISA does not invariably prove or exclude the presence of HLA antibodies, and that additional testing is required in some cases. PMID- 16712644 TI - A polymorphism in the promoter region of the CD86 (B7.2) gene is associated with systemic sclerosis. AB - Systemic sclerosis (SSc) is a connective tissue disease of unknown aetiology characterized by fibrosis of the skin and internal organs, vascular abnormalities and humoral autoimmunity. Strong T-cell-dependent autoantibody and HLA associations are found in SSc subsets. The co-stimulatory molecule, CD86, expressed by antigen-presenting cells, plays a crucial role in priming naive lymphocytes. We hypothesized that SSc, or one of the disease subsets, could be associated with single-nucleotide polymorphisms of the CD86 gene. Using sequence specific primer-polymerase chain reaction (SSP-PCR) methodology, we assessed four CD86 polymorphisms in 221 patients with SSc and 227 healthy control subjects from the UK. Haplotypes were constructed by inference and confirmed using PHASE algorithm. We found a strong association between SSc and a specific haplotype (haplotype 5), which was more prevalent in patients than in controls (29% vs 15%, OR = 2.3, chi(2) = 12, P = 0.0005). This association could be attributed to the novel -3479 promoter polymorphism; a significant difference was observed in the distribution of the CD86 -3479 G allele in patients with SSc compared to controls (43.7% vs. 32.4%, OR = 1.7, chi(2) = 12.1, P = 0.0005). TRANSFAC analyses suggest that the CD86-3479T allele contains putative GATA and TBP sites, whereas G allele does not. We assessed the relative DNA protein-binding activity of the -3479 polymorphism in vitro using electromobility gel shift assays (EMSA), which showed that the -3479G allele has less binding affinity compared to the T allele for nuclear proteins. These findings highlight the importance of co-stimulatory pathways in SSc pathogenesis. PMID- 16712646 TI - Inhibitory and activatory KIR gene frequencies in the Polish population. AB - Killer cell immunoglobulin-like receptors (KIRs) present on natural killer cells and minor subpopulations of T cells recognize class I human leukocyte antigen (HLA) molecules on the surface of target cells. Human individuals differ by the presence or absence of some KIR genes on their chromosomes (haplotypic polymorphism). As KIRs (especially two-immunoglobulin-domain-like containing, or KIR2D, molecules) are important for the outcome of tissue (particularly for haematopoietic stem cell) transplantation and possibly for pregnancy, the knowledge of KIR gene distribution in a given human population is of practical value. Therefore, we tested 175 healthy individuals from Poland for the presence or absence of these KIR genes which show haplotypic polymorphism and are expressed. Results were compared with those published for other human populations, showing close relations with other Caucasoids. PMID- 16712647 TI - Genetic diversity at the hinge region of the unique immunoglobulin heavy gamma (IGHG) gene in leporids (Oryctolagus, Sylvilagus and Lepus). AB - Unlike other species, European rabbit (Oryctolagus cuniculus) possesses only one immunoglobulin gamma class. Allelic diversity at the Ig (immunoglobulin) gamma constant region encoded by the unique IGHG (immunoglobulin heavy gamma) gene is moreover much reduced. In the European rabbit, the genetic variation at IGGH hinge region is limited to a single nucleotide substitution, which causes a Met Thr interchange at amino acid position 9 (IMGT hinge numbering). We have analysed the diversity at this region more in-depth by, (1) analysing the allelic variation in 11 breeds of domestic European rabbit (Oryctolagus cuniculus cuniculus), and (2) sequencing the gamma hinge exon in wild specimens of six species of rabbit (Oryctolagus and Sylvilagus) and hares (Lepus), including the two Oryctolagus subspecies (O. cuniculus cuniculus and O. cuniculus algirus). It appeared that among leporid species, amino acid changes occur exclusively at positions 8 and 9. However, while position 8 is occupied by either Pro or Ser residues, four different residues can occur at position 9 (Met, Thr, Pro and Leu). This variation concerns sites of potential O-glycosylation and/or proteolytic cleavage, suggesting that the underlying genetic diversity could be the outcome of selection. Preservation of the gamma hinge polymorphism in domestic stocks could therefore be important. We report here a polymerase chain reaction/restriction fragment length polymorphism protocol that has allowed the monitoring of the heterozygosity levels at the gamma hinge in 11 breeds of domestic European rabbit. PMID- 16712648 TI - Understanding the genetics of autoimmune disease: two loci that regulate late onset Addison's disease in Portuguese Water Dogs. AB - Addison's disease, an immune-mediated disorder caused by destruction of the adrenal glands, is a rare disorder of Western European populations. Studies indicate that the disorder is polygenic in nature, involving specific alleles of the CTLA-4, DRB1*04 and DQ, Cyp27B1, VDR and MIC-A and -B loci. A similar immune form of Addison's disease occurs in several breeds of domestic dog, with frequencies ranging from 1.5 to 9.0%. The high frequency of the disease in domestic dog breeds likely reflects the small number of founders associated with many breeds, subsequent inbreeding, and the frequent use of popular sires. The Portuguese Water Dog (PWD) is a significantly affected breed. An analysis of 11,384 PWDs surveyed between 1985 and 1996 suggests a breed-specific disease incidence of 1.5%. As with humans, the disease is typically of late onset. This study involves a genetic comparison of Addison's disease in the PWD to the analogous disease in humans. The study is facilitated by the existence of complete pedigrees and a relatively high degree of inbreeding among PWDs. The breed originated from 31 founders, with 10 animals responsible for 90% of the current gene pool. We describe, specifically, the identification of two disease associated loci, on Canis familiaris (CFA) chromosomes CFA12 and 37, which are syntenic with the human DRB1 histocompatibility locus alleles HLA-DRB1*04 and DRB1*0301, and to a locus for immunosuppression syntenic with CTLA-4. Strong similarities exist therefore in the complex genetic background of Addison's disease in humans and in the PWD. With the completion of the canine and human genome sequence, the purebred dog is set to become an important comparative model for Addison's as well as other human immune disorders. PMID- 16712649 TI - HLA class II gene polymorphism in Parsees and Zoroastrians of Iran. AB - Extensive polymorphism of the HLA genes in different ethnic groups has been used as an invaluable tool for anthropological studies. In this study, HLA-DRB1, DQA1 and DQB1 allele frequencies and haplotypes were determined in 72 Parsees and 65 Zoroastrians living in Iran. The predominant DRB1 allele was *1103 = 4 in Parsees and *0701 in Zoroastrians. DQA1*0501 was the most common alleles in both spopulations. The most frequent DQB1 allele was *0301 in Parsees and *0201 in Zoroastrians. DRB1*1103 = 4-DQA1*0501-DQB1*0301 and DRB1*0701-DQA1*0201-DQB1*0201 were the most prevalent haplotypes in Parsees and Zoroastrians, respectively. Significant deviation from Hardy-Weinberg equilibrium was observed in DQA1 and DQB1 loci of Zoroastrians. The former locus also departed from neutrality due to balancing selection. All pairs of the studied loci in this study showed significant linkage disequilibrium. Analysis of molecular variance indicated that the main variation was confined to individuals within the studied populations. Neighbour-joining tree based on Nei's genetic distances according to DRB1 and DQB1 allele frequencies showed that Parsees and Zoroastrians of Iran were located in the same cluster of the phylogenetic tree. Furthermore, Zoroastrians of Iran and Pakistan are very close to each other. This study will serve as a reference for further anthropological studies when the HLA profile of all ethnic groups of Iran is investigated. PMID- 16712650 TI - Cytokine typing: SNP allele frequencies in the Iranian population. AB - Single nucleotide polymorphisms within the promoter or other regulatory sequences of cytokine genes were evaluated and compared between an Iranian population and populations of different ethnic/geographical background. In 40 healthy Iranian subjects, cytokine typing was performed by polymerase chain reaction-sequence specific primers assay. Allele frequencies were calculated and compared with those of other populations. PMID- 16712651 TI - A single amino acid exchange shifts the serological reactivity of the novel HLA B*4442 allele product from HLA-B44 to HLA-B21. AB - A novel HLA-B (human leukocyte antigen-B) allele, HLA-B*4442, was identified both in a Czech patient with leukaemia and in his mother. The presence of a novel allele was initially suspected because conflicting results were obtained by serological and DNA typing techniques. The HLA typing using the polymerase chain reaction-sequence-specific primers (PCR-SSP) at the two-digit level indicated an allele belonging to the HLA-B*44 group, whereas serological typing indicated HLA B21. Typing with PCR-sequence-specific oligonucleotides (PCR-SSO) resulted in a unique reaction pattern that could not be assigned to a known allele, PCR-SSP typing at the four-digit level did not match any known B*44 allele, either. The sequencing-based typing of the HLA-B locus then revealed the novel B*4442 allele that is identical with B*4405 except a single C-->G nucleotide exchange at position 572. This exchange results in an amino acid substitution from serine to tryptophan at position 167 of the expressed HLA-B protein. The B21 serological reactivity of the novel B*4442 allele product was confirmed by employing an additional serological panel of typing sera. Our findings support previous reports claiming that serine at the position 167 in the alpha-2 domain of the HLA B protein is a major determinant of the HLA-B44(12) serological epitope. PMID- 16712652 TI - Genetic polymorphisms of chitotriosidase in Caucasian children with bronchial asthma. AB - In humans, two types of chitinases have been identified: chitotriosidase I (CHIT1) and acid mammalian chitinase (AMCase). They are enzymes that cleave chitin, a polysaccharide contained in many different human parasites. So far, only little is known about their function in human and especially in human diseases. Recently we have described association of polymorphisms of AMCase with bronchial asthma in a pediatric population. In this study we were interested in whether CHIT1 is also involved in the genetics of asthma. The amino acid variants Gly102Ser and Ala442Gly, as well as a 24 bp duplication within CHIT1, were typed by means of restriction fragment length polymorphisms on 322 children with asthma and 270 randomly chosen adult controls. Statistical analyses made use of the Armitage's trend test; haplotypes were calculated by FAMHAP and FASTEHPLUS. The amino acid variants showed no association with bronchial asthma. The 24 bp duplication, previously shown to completely demolish CHIT1 activity, was also evenly distributed between asthmatics and controls. Finally, the haplotype showed no association with the disease. We conclude from our results that CHIT1 does not play a major role in the development of bronchial asthma in Caucasian children. The results might also imply that the two human chitinases that have been identified so far have quite distinct functions in human diseases even though they have the same substrate. PMID- 16712653 TI - Identification of Bruton tyrosine kinase mutations in 12 Chinese patients with X linked agammaglobulinaemia by long PCR-direct sequencing. AB - X-linked agammaglobulinaemia (XLA) is an immunodeficiency caused by Bruton tyrosine kinase (BTK) gene mutations. The disease is characterized by recurrent bacterial infections and profound hypogammaglobulinemia with marked reduction or lack of mature B-cells in the peripheral blood. Molecular characterization of BTK gene provides an opportunity for definitive diagnosis of XLA patients, especially for those with atypical phenotype resulting in a milder or late-onset form of the disease. The diagnosis allows accurate carrier detection with subsequent genetic counselling and prenatal diagnosis. In this study, long polymerase chain reaction (PCR)-direct sequencing analysis of the BTK gene in 12 unrelated Chinese XLA patients had been performed. Eight recurrent mutations and four novel mutations were identified. This is the first report of Chinese cases from three different East Asia regions together, including Hong Kong, Singapore and mainland China. Future clinical and genetic information from the undiagnosed Chinese XLA patients may provide insight into the genotype-phenotype correlations of BTK gene. PMID- 16712654 TI - Dinucleotide repeat polymorphism in intron II of human Toll-like receptor 2 gene and susceptibility to rheumatoid arthritis. AB - Human Toll-like receptors (TLRs) participate in innate immune response and signal the activation of adaptive immunity. The presence of a functional intronic polymorphism consisting of guanine-thymine repeats in TLR2 gene was recently reported. Here, we investigated a dinucleotide repeat polymorphism in intron II of TLR2 in Korean patients with rheumatoid arthritis (RA). The numbers of guanine thymine [(GT)(n)] repeats in intron II of the TLR 2 gene were counted in 183 patients with RA and in 148 healthy controls, using the gene scanning technique. We classified alleles into two subclasses for further analysis, 12-16 GT repeats (S allele) and 17-28 repeats (L allele). By subgroup analysis, we also examined whether the S allele is associated with the presence of shared epitope (SE), rheumatoid factor (RF), joint erosion and extra-articular complications. S-allele frequency was significantly increased in patients with RA than in healthy controls [30.3% vs. 23.0%, P = 0.03, or 1.46, 95% confidence interval (CI) 1.03 2.07], and genotypes containing S alleles were more frequent in patients with RA than in healthy controls (54.4% vs. 46.5%. P = 0.04, or 1.57, 95% CI 1.01-2.42). A skewed S-allele distribution was not found to be related to the presence of SE. Subgroup analysis showed no genotypic or allele frequency differences between patients with/without RF, joint erosion, or extra-articular complications. Genotype containing shorter GT repeats in intron II of the TLR2 gene may confer susceptibility to RA in Koreans. PMID- 16712655 TI - CTLA-4 gene polymorphisms and natural soluble CTLA-4 protein in psoriasis vulgaris. AB - CTLA-4 molecule is an important inhibitor of T-lymphocyte activation. Several single nucleotide polymorphisms (SNPs) in the CTLA-4 gene were found, and their associations with many human diseases were described. So far, however, such studies have not been performed in psoriasis vulgaris in Caucasoids. Therefore, we examined the distribution of three CTLA-4 SNPs: -1147C/T, -318C/T and +49 A/G in 116 patients with psoriasis vulgaris and 123 healthy blood donors using the polymerase chain reaction-restriction fragment length polymorphism method. For all three SNPs, the frequencies of alleles, genotypes and three-point haplotypes were very similar in patients and controls, suggesting no contribution of these genetic variants to psoriasis. PMID- 16712658 TI - Association between the 1188 A/C polymorphism in the human IL12B gene and Th1 mediated infectious diseases. PMID- 16712660 TI - Anti-Mullerian hormone (AMH) in female reproduction: is measurement of circulating AMH a useful tool? AB - Anti-Mullerian hormone (AMH) is a dimeric glycoprotein, a member of the transforming growth factor (TGF) superfamily. It is produced exclusively in the gonads and is involved in the regulation of follicular growth and development. In the ovary AMH is produced by the granulosa cells of early developing follicles and seems to be able to inhibit the initiation of primordial follicle growth and FSH-induced follicle growth. As AMH is largely expressed throughout folliculogenesis, from the primary follicular stage towards the antral stage, serum levels of AMH may represent both the quantity and quality of the ovarian follicle pool. Compared to other ovarian tests, AMH seems to be the best marker reflecting the decline of reproductive age. AMH measurement could be useful in the prediction of the menopausal transition. It could also be used to predict poor ovarian response and possibly the prognosis of in vitro fertilization (IVF) cycles. AMH has been shown to be a good surrogate marker for polycystic ovary syndrome (PCOS). Finally, its use as a marker for granulosa cell tumours has been proposed. A clearer understanding of its role in ovarian physiology may help clinicians to find a role for AMH measurement in the field of reproductive medicine. PMID- 16712661 TI - Hyperthyroidism is characterized by both increased sympathetic and decreased vagal modulation of heart rate: evidence from spectral analysis of heart rate variability. AB - OBJECTIVE: The clinical manifestations of hyperthyroidism resemble those of the hyperadrenergic state. This study was designed to evaluate the impact of hyperthyroidism on the autonomic nervous system (ANS) and to investigate the relationship between serum thyroid hormone concentrations and parameters of spectral heart rate variability (HRV) analysis in hyperthyroidism. DESIGN AND PATIENTS: Thirty-two hyperthyroid Graves' disease patients (mean age 31 years) and 32 sex-, age-, and body mass index (BMI)-matched normal control subjects were recruited to receive one-channel electrocardiogram (ECG) recording. MEASUREMENTS: The cardiac autonomic nervous function was evaluated by the spectral analysis of HRV, which indicates the autonomic modulation of the sinus node. The correlation coefficients between serum thyroid hormone concentrations and parameters of the spectral HRV analysis were also computed. RESULTS: The hyperthyroid patients revealed significant differences (P < 0.001) compared with the controls in the following HRV parameters: a decrease in total power (TP), very low frequency power (VLF), low frequency power (LF), high frequency power (HF), and HF in normalized units (HF%); and an increase in LF in normalized units (LF%) and in the ratio of LF to HF (LF/HF). After correction of hyperthyroidism in 28 patients, all of the above parameters were restored to levels comparable to those of the controls. In addition, serum thyroid hormone concentrations showed significant correlations with spectral HRV parameters. CONCLUSIONS: Hyperthyroidism is in a sympathovagal imbalanced state, characterized by both increased sympathetic and decreased vagal modulation of the heart rate. These autonomic dysfunctions can be detected simultaneously by spectral analysis of HRV, and the spectral HRV parameters could reflect the disease severity in hyperthyroid patients. PMID- 16712662 TI - Lithium as adjuvant to radioiodine therapy in differentiated thyroid carcinoma: clinical and in vitro studies. AB - OBJECTIVE: Lithium has been reported to increase radioactive iodine (RaI) doses in benign thyroid disease and in differentiated thyroid carcinoma (DTC). It is not known whether lithium influences the outcome of RaI therapy in DTC. We therefore studied the clinical effects of RaI without and with lithium carbonate in patients with proven metastatic DTC. Controversy also exists on the mechanism by which lithium increases RaI dose in DTC. We performed an in vitro study specifically aimed at examining the effects of lithium on the sodium iodide symporter (NIS). DESIGN: In a clinical study, 12 patients were selected with metastases of DTC who had received previous RaI therapy without lithium (control) that had not influenced tumour progression, despite RaI accumulation in metastases. The patients received 1200 mg lithium carbonate/day followed by 6000 MBq RaI. Outcome parameters were RaI uptake, serum thyroglobulin (Tg) levels and radiological dimensions of metastases compared between RaI with lithium and control. In an in vitro study, iodide uptake was studied in the benign rat thyroid cell line FRTL-5, in the polarized non-thyroid MDCK cell line, stably transfected with human sodium iodide symporter (hNIS) to study the effects of lithium on NIS in a non-thyroid background, and the human follicular thyroid carcinoma cell line FTC133-hNIS to study lithium effects in a background of DTC. Lithium chloride (LiCl) was added in concentrations up to 2 mM for 0-48 h. Both steady-state iodide uptake (30 min) and initial rate (2 min) were studied using a specific activity of 100 mCi/mmol I, the latter experiment to determine lithium effects on substrate dependency. Iodide efflux studies were performed as well. RESULTS: Despite an increased uptake of RaI in seven patients, no beneficial effect of RaI with lithium was observed on the clinical course as assessed by serum Tg measurements and radiographically. In the in vitro studies, no effects of LiCl on iodide uptake or efflux were observed. CONCLUSIONS: The addition of lithium to RaI did not have any beneficial effects on the clinical course in 12 patients with metastatic DTC. No beneficial effects of lithium on iodide uptake were observed in vitro. Therefore, the clinical value of lithium in DTC remains subject to debate. PMID- 16712663 TI - Bone metabolism markers predict increase in bone mass, height and sitting height during puberty depending on the VDR Fok1 genotype. AB - OBJECTIVES: Longitudinal growth and bone mass accumulation are two important phenomena during puberty, resulting in attainment of peak bone mass (PBM) and final height. They are thought to be under strong genetic control, with the vitamin D receptor (VDR) gene being among the candidate genes. Bone metabolism markers are reported to be good predictors of longitudinal growth and bone mass increase. The purpose of this longitudinal study was to evaluate whether bone metabolism markers predict bone mass and height increase differently according to Fok1 VDR genotype throughout puberty in healthy adolescents. PATIENTS AND MEASUREMENTS: At the start of the study 130 girls were aged 10.8 (range 8.5-12.8) years and 125 boys were aged 11.8 (range 9.4-14.6) years at the first visit. Markers of bone formation and bone resorption were measured at the first visit, as well as height and sitting height (SH), and bone mineral content (BMC) of the lumbar spine, femur, arm and total body. All measurements were repeated after 2 years. RESULTS: A higher BMC of the femur, distal arm and total body was found in ff boys at the first visit, which was not related to higher levels of bone metabolism markers in this group. In girls, no differences between genotypes were seen in BMC (increase). However, concentrations of markers of bone formation [alkaline phosphatase (AP), bone-specific alkaline phosphatase (BAP), procollagen aminoterminal propeptide (PINP)] and bone resorption [type I carboxyterminal telopeptide (ICTP)] were higher in ff girls. Regression coefficients between bone metabolism markers and bone mass increase differed according to genotype and sex. A similar pattern was found for height and SH (increase), the latter as a representative of growth of the axial skeleton, mainly consisting of cancellous bone. CONCLUSIONS: Our data suggest that the predicting capacities of bone metabolism markers on bone mass (increase), height and SH (increase) are genotype dependent. Their use as predictors of final height or PBM therefore remains questionable without knowing the genotype. PMID- 16712664 TI - Peripheral fibrinolytic markers, soluble adhesion molecules, inflammatory cytokines and endothelial function in hypopituitary adults with growth hormone deficiency. AB - BACKGROUND: Increased mortality due to cardiovascular disease has been described in adult patients with untreated GH deficiency (GHD). GH replacement has been demonstrate to improve vascular reactivity and reverse early atherosclerotic changes in adults with GHD. OBJECTIVE: Assessment of fibrinolytic markers, soluble adhesion molecules, inflammatory cytokines and endothelial function in hypopituitary adults with GHD at baseline and 1 year after GH replacement therapy. METHODS: We studied 10 patients with GHD (five men and five women; aged 45.6 +/- 10.4 years) at baseline and 1 year after GH replacement therapy compared with a control group (nine men and 16 women) matched for age and body mass index (BMI). All subjects, patients and controls, were life-long nonsmokers, normotensive and nondiabetic. The following variables were recorded: anthropometric and body composition variables, serum concentrations of glucose, insulin and C-peptide; thrombin anti-thrombin (TAT) fragments and fibrin degradation product D-dimer, which were determined by an enzyme-linked immunosorbent assay (ELISA); IGF-I by radioimmunoassay (RIA); C-reactive protein (CRP) by highly sensitive immunonephelometry; E-selectin, P-selectin, soluble intercellular cell adhesion molecule-1 (sICAM-1), soluble vascular cell adhesion molecule-1 (sVCAM-1), interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) by ELISA. The assessment of endothelial function in vivo was measured with a Doppler device. RESULTS: Patients with GHD without GH substitution had higher hip/waist ratio and body fat than controls. Insulin, C-peptide and triglyceride concentrations were also higher. Our results demonstrated no difference in fibrinogen and in TAT fragment concentrations among patients and controls. E-selectin concentrations were higher in patients than in controls (26.1 +/- 11 vs. 10.7 +/- 6.2 microg/l, P = 0.0001). P-selectin, sICAM-1, sVCAM 1, IL-6, MCP-1 and CRP were similar in the two groups. Vascular reactivity and carotid intima-media thickness (IMT) were also similar in patients and controls. After 1 year of GH treatment we found no changes in biochemical parameters, fibrinolytic markers, soluble adhesion molecules, inflammatory cytokines and endothelial function. CONCLUSION: Adults with GHD show some subtle changes in soluble adhesion molecules but our data suggest no beneficial effects of GH over these markers in relationship to endothelial function. Factors other than GH treatment, such as differences in age, degree of obesity, the presence of diabetes mellitus and arterial hypertension or tobacco consumption, could explain the observed increase in markers of vascular risk in GH-deficient patients. PMID- 16712665 TI - Longitudinal assessment of insulin sensitivity in pregnancy. Validation of the homeostasis model assessment. AB - OBJECTIVE: To validate the use of the homeostasis model assessment (HOMA) for measurement of insulin sensitivity in obese women during gestation and the postpartum period. DESIGN: Three consecutive measurements of insulin resistance (IR) were performed during and after pregnancy to compare the homeostasis model assessment insulin sensitivity index (HOMA-IR) to glucose utilization rates obtained during hyperinsulinaemic euglycaemic clamps (G(Rd)). PATIENTS: Six obese women (mean second trimester BMI = 30.4 kg/m(2)) with normal glucose tolerance were studied during the second and third trimesters of pregnancy and once in the postpartum period. Thus, there were a total of 18 measurements for analysis. RESULTS: Correlations between the rate of glucose disappearance (G(Rd)) (the gold standard) and the HOMA-derived metabolic parameters of insulin sensitivity were significant, with a multiple R(2) of 43.5% (P = 0.003). However, when controlling for variations between patients using dummy variables, we observed that one patient differed from the other five in the relationship between G(Rd) and HOMA IR. Applying this regression we obtained a R(2) of 72.6% (P < 0.001). When the regression constant was omitted, we observed that the individual trends during pregnancy and postpartum in two patients differed statistically from the other patients between the two assessments, and we obtained a multiple R(2) of 97.3% (< 0.001). CONCLUSIONS: HOMA estimation of insulin resistance is appropriate for use during both the second and third trimesters of pregnancy and postpartum in obese women with normal glucose tolerance. It lacks sensitivity for the evaluation of individuals, where more precise measures of insulin sensitivity should be utilized. PMID- 16712666 TI - Hyperandrogenism in carriers of CYP21 mutations: the role of genotype. AB - OBJECTIVE: It has been hypothesized that carriers of CYP21 mutations are at increased risk of developing clinically evident hyperandrogenism. In the present study, we assessed the risk for symptoms of androgen excess in carriers of CYP21 gene mutations and the effect of different mutations on phenotype. DESIGN: All subjects underwent clinical evaluation, ACTH stimulation testing, and molecular analysis of the CYP21 gene. PATIENTS: One hundred and eighty-seven subjects were included in the study. Five groups were defined according to genotype: A, 38 patients with classical-congenital adrenal hyperplasia (CAH); B, 16 patients with non classical CAH; C, 66 family member carriers; D, 24 children and adolescents with symptoms suggestive of hyperandrogenism who were found to be heterozygous and E, 43 subjects with androgen excess symptoms who had normal genotype. MEASUREMENTS: Cortisol and 17-hydroxyprogesterone (17OHP) were measured basally and 60 min after ACTH stimulation. Analysis of seven common mutations of the CYP21 gene, which collectively cover 95% of the mutations in the Israeli population, was performed. The hormonal results were correlated with the genotype. RESULTS: The symptomatic carriers had a significantly higher rate of the mild mutation V281L compared with the family members (58% vs. 22%) and significantly higher levels of ACTH-stimulated 17OHP (mean 37.2 and 21 nmol/l, respectively; P = 0.025). Higher values of peak 17OHP levels were found in the carriers of the V281L mutation compared with carriers of other mutations (P = 0.025). Stimulated 17OHP levels were significantly higher in carriers compared with normal genotype subjects (P < 0.0001). CONCLUSIONS: Our findings are the first to show that there is increased risk of androgen excess in carriers of CYP21 mutations and that the risk is related to genotype. Carriers for the mild, V281L mutation, are at higher risk of symptoms of androgen excess than carriers of the severe mutations. It appears that the variable phenotypes can be at least partially attributed to the degree of impairment of enzyme activity in different mutations. The severe consequences of hyperandrogenism in some of the carrier subjects indicate the need for long-term follow-up in these patients. PMID- 16712667 TI - Circulating osteoprotegerin is correlated with lipid profile, insulin sensitivity, adiponectin and sex steroids in an ageing male population. AB - OBJECTIVE: The relationship between osteoprotegerin (OPG) and lipid profile, insulin sensitivity, adipocytokines and sex steroids has been poorly studied and subject to controversy. The purpose of this study was to look at the correlates of OPG in an elderly male population. DESIGN: One hundred and fifty-one nondiabetic, elderly Lebanese men (age range 50-83) were recruited in this cross sectional study based on voluntary enrolment. MEASUREMENTS: In all the subjects, serum OPG levels were measured and related to clinical parameters (age, waist, body mass index (BMI), systolic and diastolic blood pressure), as well as to metabolic and hormonal parameters. The following fasting laboratory measurements were performed: plasma glucose and insulin levels, total cholesterol, triglycerides and HDL cholesterol, adiponectin, leptin, as well as sex steroids (testosterone, SHBG, free androgen index, ooestradiol, DHEAS), GH and IGF-1. QUICKI index was calculated as a measure of insulin sensitivity. RESULTS: OPG levels were significantly correlated with age (r = 0.28, P < 0.0001) but not with BMI, waist, systolic or diastolic blood pressure. There was a trend towards higher OPG levels in subjects without, compared to subjects with the metabolic syndrome (3.58 +/- 1.28 vs. 3.26 +/- 1.04 pmol/l, P = 0.09). OPG was negatively correlated with fasting glucose and triglyceride levels (r = -0.18, P = 0.031 and r = -0.19, P = 0.02, respectively) and positively correlated with the QUICKI index (r = 0.17, P = 0.033), HDL cholesterol (r = 0.21, P = 0.009) and adiponectin levels (r = 0.27, P = 0.001). No significant correlations were reported with total or LDL cholesterol levels and with leptin levels. After adjustment for age, OPG is still correlated with triglycerides (r = -0.19, P = 0.02), glucose (r = -0.21, P = 0.011) and adiponectin (r = 0.19, P = 0.02). Finally, OPG was positively associated with SHBG (r = 0.31, P < 0.001) and negatively associated with free androgen index (r =-0.346, P < 0.001); both correlations persisted after adjustment for age (r = 0.21, P = 0.009 and r = 0.23, P = 0.005, respectively). No significant correlation was found between OPG and oestradiol levels while a weak negative correlation was demonstrated with DHEAS (r = -0.18, P = 0.025). Also, no significant correlation was found between OPG and GH or IGF-1 values. In a multiple regression analysis with a stepwise model, the main determinants of OPG were free androgen index and adiponectin (P < 0.0001 and P = 0.015, respectively). CONCLUSION: Our results show that circulating OPG levels are favourably associated with some components of the metabolic syndrome. Also, for the first time, an association between OPG and adiponectin is described. Finally, the negative correlation we found between OPG and free androgen index may suggest a potential role of OPG in the increase in cardiovascular disease related to ageing and sex steroid deficiency. PMID- 16712668 TI - Occurrence of the Cys611Tyr mutation and a novel Arg886Trp substitution in the RET proto-oncogene in multiple endocrine neoplasia type 2 families and sporadic medullary thyroid carcinoma cases originating from the central region of Portugal. AB - OBJECTIVE: Medullary thyroid carcinoma (MTC) occurs both sporadically and in the context of autosomal dominantly inherited multiple endocrine neoplasia type 2 (MEN2) syndromes: MEN2A, MEN2B, and familial medullary thyroid carcinoma (FMTC), which are caused by activating germline mutations in the RET proto-oncogene. The aim of this study was to characterize the RET mutational spectrum in MEN2 families and apparently sporadic MTC (AS-MTC) cases originating from the central region of Portugal. SUBJECTS AND METHODS: We studied a total of 82 individuals (64 affected and 18 family members), comprising five MEN2 families (four MEN2A and one MEN2B), as well as 53 AS-MTC cases. RET germline mutations were screened using PCR-DNA sequencing, SSCP and RFLP. The haplotypes associated with recurrent mutations were determined by fragment analysis of microsatellite markers, and by RFLP, in the case of intragenic polymorphisms. RESULTS: Frequency of the Cys611Tyr (TGC-TAC) mutation was significantly increased in this region of Portugal, due to the fact that three apparently unrelated MEN2A/FMTC families, out of the five in which mutations were identified, harboured this specific mutation. Haplotype analysis revealed that a common haplotype was shared between two of these three families. We have also characterized a novel RET mutation, Arg886Trp, located in the tyrosine kinase domain, which was found in an AS-MTC case. CONCLUSIONS: There are regional specificities in the relative frequency of RET mutations, which are consistent with a cluster-like distribution of specific disease-causing mutations, as a result of the inheritance of a shared haplotype. These data, along with the finding of a novel RET mutation (Arg886Trp), have important implications towards facilitating and improving the molecular diagnosis of hereditary MTC on a regional basis. PMID- 16712669 TI - Effect of obesity and morbid obesity on the growth hormone (GH) secretion elicited by the combined GHRH + GHRP-6 test. AB - OBJECTIVE: Obesity is characterized by low basal levels of growth hormone (GH) and impeded GH release. However, the main problem arises in the diagnosis of GH deficiency in adults, as all accepted cut-offs in the diagnostic tests of GH reserve are no longer valid in obese subjects. In this work, the role of obesity in the GH response elicited by the GHRH + GHRP-6 test was assessed in a large population of obese and nonobese subjects. PATIENTS: GHRH + GHRP-6-induced GH peaks were evaluated in 542 subjects. One hundred and five were healthy obese, 50 were morbid obese, and 261 were nonobese (both normal weight and overweight). One hundred and seventy-six GH-deficient patients (obese and nonobese) were also studied. RESULTS: A regression analysis of the 366 subjects with normal pituitary function indicated that adiposity had a negative effect on the elicited GH peak (r = -0.503, P < 0.0001). A receiver operating characteristic (ROC) curve analysis showed that in subjects with a BMI < or =35, the currently accepted cut offs of the GHRH + GHRP-6 test (GH peaks > or =20 microg/l: normal secretion; GH peaks < or =10 microg/l: GH deficiency), were fully operative. However, in subjects with a BMI > 35, normality was indicated by GH peaks > or =15 microg/l and GH deficiency by peaks < or =5 microg/l (1 microg/l = 2.6 mU/l). CONCLUSIONS: This study confirms: (a) that the combined provocative test is adequate to separate normal and GH-deficient subjects; (b) the negative effect of obesity on GH secretion; (c) that obesity accounts for 25% of the reduction of GH release; and (d) that present cut-off values are applicable to normal weight, overweight and grade I obesity subjects, whereas in obese subjects with a BMI exceeding 35, all the normative limits of the GHRH-GHRP +6 test must be reduced by 5 microg/l. PMID- 16712670 TI - Insulin resistance and the metabolic syndrome in obese French children. AB - OBJECTIVE: To estimate the frequency of the metabolic syndrome (MS) and of the insulin resistance syndrome (IRS) in overweight or obese French children and to determine the risk factors. DESIGN, PATIENTS AND METHODS: A total of 308 overweight and obese children [166 girls, 142 boys, aged 7-17 years; median body mass index (BMI) 4.7 standard deviation (SD) (Q1-Q3: 3.9-5.8) adjusted for age and sex] were included. The frequency of the MS was assessed with the National Cholesterol Education Program (NCEP) Adult Treatment Panel III (ATP III) criteria and the frequency of the IRS with World Health Organization (WHO) criteria. RESULTS: The overall frequency of MS and IRS was 15.9% and 42.5%, respectively. The most common component, after abdominal obesity (95.8%) and IR (71.8%), was elevated systolic blood pressure (28.6%). The frequency of glucose tolerance disorders was low (3.6%). The frequency of MS was independently influenced by homeostatic model assessment (HOMA) (P = 0.06) and waist-to-hip ratio (P = 0.09), whereas the frequency of IRS was influenced by adiposity (degree of obesity: P = 0.02; waist-to-hip ratio: P = 0.05), puberty (P = 0.05) and mother's BMI (P = 0.01). Ethnicity had no effect on either MS or IRS. CONCLUSIONS: Metabolic complications and IR are frequent in overweight and obese children whereas the frequency of glucose tolerance disorders is very low. IRS is more prevalent than MS, indicating a major role of IR, which could precede the other metabolic complications in obese children. IRS is a relevant marker for the risk of type 2 diabetes (T2D) and cardiovascular complications in obese European children. PMID- 16712671 TI - Associations between plasma adiponectin concentrations and liver histology in patients with nonalcoholic fatty liver disease. AB - OBJECTIVES: To explore associations between plasma adiponectin concentrations and liver histology in patients with nonalcoholic fatty liver disease (NAFLD). DESIGN AND PATIENTS: In a cross-sectional study, we enrolled 60 consecutive NAFLD patients and 60 age-, sex- and body mass index (BMI)-matched healthy controls. MEASUREMENTS: NAFLD (by liver biopsy), plasma adiponectin concentrations, insulin resistance (by homeostasis model assessment, HOMA-IR) and metabolic syndrome (MetS) features. RESULTS: NAFLD patients had a marked decrease in plasma adiponectin concentration (6.1 +/- 2.8 vs. 13.6 +/- 3.8 microg/ml, P < 0.001) compared with matched controls. MetS, as defined by the Adult Treatment Panel III (ATP III) criteria, and its individual components were more frequent among NAFLD patients. The marked differences in adiponectin concentrations that were observed between the groups were little affected by adjustment for age, sex, BMI, HOMA-IR score and MetS components. Notably, decreased adiponectin levels were closely associated with the degree of hepatic steatosis, necroinflammation and fibrosis (P < 0.001 for all) among NAFLD patients. By logistic regression analysis, low adiponectin levels independently predicted hepatic steatosis [odds ratio (OR) 2.3, 95% confidence interval (CI) 1.5-5.8, P < 0.001] and necroinflammation (OR 3.1, 95% CI 1.9-7, P < 0.001), but not fibrosis (P = 0.07), after adjustment for age, sex, BMI, HOMA-IR and MetS components. CONCLUSIONS: NAFLD patients have markedly lower plasma adiponectin concentrations than control subjects. Low adiponectin levels are strongly associated with the severity of liver histology, thus further supporting the hypothesis that adiponectin might be involved in the development of NAFLD. PMID- 16712672 TI - Plasma ghrelin levels of gastrectomized and vagotomized patients are not affected by glucose administration. AB - BACKGROUND: Ghrelin is a brain-gut peptide with GH-releasing and appetite inducing activities, secreted mainly by the stomach. Circulating ghrelin concentrations fall rapidly after nutrient ingestion as well as after oral and intravenous glucose challenge. A number of gut hormones including ghrelin require an intact vagal system, which has been hypothesized to have a major role in initiating the postprandial fall in ghrelin levels. AIM: We aimed to investigate the effect of oral glucose challenge on ghrelin secretion in gastrectomized (GASTRX) and vagotomized patients. DESIGN: Interventional study. PATIENTS: Six GASTRX-vagotomized patients and 11 healthy sex- and body mass index (BMI)-matched subjects. METHODS: An oral glucose tolerance test (OGTT) was performed in all subjects. At baseline, circulating plasma total ghrelin, serum glucose, insulin and GH levels were measured. Serum glucose, insulin, GH and plasma ghrelin levels were determined every 30 min for 2 h. RESULTS: Plasma ghrelin levels at baseline were reduced by 55% in GASTRX-vagotomized patients compared to the control group (P < 0.01). In control subjects, plasma ghrelin levels decreased significantly during the OGTT whereas in GASTRX-vagotomized patients no reduction was registered (26.4 +/- 2.8% vs. 5.5 +/- 3.4%). The OGTT revealed a significantly greater increase in circulating glucose levels and serum insulin levels while GH response was not different in GASTRX-vagotomized patients compared to control subjects. CONCLUSIONS: Our data show that circulating ghrelin levels in GASTRX and vagotomized patients were not suppressed after oral glucose administration, unlike control subjects, suggesting that this effect could be due, at least in part, to the lack of contribution of the vagal nervous system to the regulation of ghrelin. PMID- 16712673 TI - The relationship between four single nucleotide polymorphisms in the promoter region of the osteoprotegerin gene and aortic calcification or coronary artery disease in Koreans. AB - OBJECTIVE: Osteoprotegerin (OPG) is a recently identified cytokine that acts as a decoy receptor for the receptor activator of the NF-kappaB ligand (RANKL). OPG has been shown to be an important inhibitor of osteoclastogenesis and arterial calcification in animal models. OPG has been proposed as a link molecule between osteoporosis and arterial calcification, but the relationship between the OPG gene and the cardiovascular system in human populations is unclear. Thus, the aim of this study was to investigate the relationship between OPG gene polymorphisms and aortic calcification or coronary artery disease in Koreans. DESIGN AND PATIENTS: Genotyping of four polymorphisms, A163G, G209A, T245G and T950C, in the promoter region of the OPG gene was performed in 251 healthy Korean women (mean age 51.3 +/- 6.9 years) and in a second study population consisting of 100 patients who underwent coronary angiography (mean age 57.0 +/- 11.9 years), by allelic discrimination using the 5' nuclease polymerase chain reaction assay. Cardiovascular risk factors and serum OPG levels were measured and aortic calcification in thoracic and abdominal aorta was examined by simple radiological methods. RESULTS: In the first study population, the prevalence of aortic calcification increased significantly as the subjects grew older. The frequencies of mutant alleles were significantly higher in the subjects with aortic calcification compared with those without aortic calcification in G209A and T950C polymorphisms, although these significances were lost after adjustment for age. No significant relationship was found between OPG gene polymorphisms and serum OPG levels or cardiovascular risk factors. In the second study group, there were no associations between OPG promoter genotypes and aortic calcification, serum OPG levels, or coronary artery disease. CONCLUSIONS: We observed that the four polymorphisms in the promoter region of the OPG gene were not associated with aortic calcification or coronary artery disease in Koreans. Further studies are needed to clarify this relationship. PMID- 16712674 TI - Impairment of endothelium-dependent arterial dilation in Hashimoto's thyroiditis patients with euthyroidism. AB - OBJECTIVE: Recent studies have shown that immune responses contribute to atherosclerosis, and endothelial dysfunction is an important early event in atherogenesis. The aim of this study was to investigate the alteration of endothelial function in Hashimoto's thyroiditis (HT) patients with euthyroidism. METHODS: Study subjects included 28 female HT patients with euthyroidism, 23 female HT patients with hypothyroidism, and 22 healthy women. High-resolution ultrasound was used to measure brachial artery diameter at rest, after reactive hyperaemia and after sublingual glyceryltrinitrate (GTN). RESULTS: Flow-mediated arterial dilation in HT patients with euthyroidism was significantly lower (3.88%) than in controls (4.98%, P = 0.000) and higher than in HT patients with hypothyroidism (3.26%, P < 0.001). Flow-mediated arterial dilation among HT patients with hypothyroidism was significantly lower than that in controls (P = 0.000). GTN-induced arterial dilation, baseline vessel size, and baseline blood flow were not significantly different among the three groups (P > 0.05). On multiple regression analysis, anti-thyroid peroxidase antibody (TPO-Ab), TSH, free T3, low density lipoprotein cholesterol (LDL-C) and lipoprotein (a) [Lp(a)] were found to be significant factors associated with endothelium-dependent arterial dilation. CONCLUSION: Endothelial dysfunction exists in HT patients with euthyroidism. Autoimmune reactivity and an elevated Lp(a) level might be responsible for the endothelial dysfunction. PMID- 16712675 TI - Interrelated overexpression of endothelial and inducible nitric oxide synthases, endothelin-1 and angiogenic factors in human papillary thyroid carcinoma. AB - OBJECTIVE: Nitric oxide (NO) and endothelin-1 (ET-1) are involved in carcinogenesis. Overexpression of the ET-1 axis has been demonstrated in papillary thyroid carcinoma (PTC). This study investigated the expression of NO synthases (NOS) and their relationship with expression of ET-1 and angiogenic markers in PTC. DESIGN AND PATIENTS: Expression of NOS, angiogenic markers [vascular endothelial growth factor (VEGF), angiopoietin-1 and angiopoietin-2] and their receptors was studied in surgical thyroid samples obtained from 22 patients aged 15-68 years. Three groups were constituted: normal thyroid (n = 5), Hashimoto's thyroiditis (n = 9) and PTC (n = 8). RESULTS: Immunohistochemistry disclosed NOS2 and NOS3 immunoreactivity in PTC cells, the percentage of positive cells being greater than normal (P < 0.02). Real-time quantitative polymerase chain reaction (RTQ-PCR) showed that NOS2 and NOS3 mRNA levels were, respectively, increased (P < 0.02) by 2.6 +/- 0.6 and 4.2 +/- 1.1 times in PTC. RTQ-PCR demonstrated that VEGF, its receptors VEGFR-1 and VEGFR-2, and angiopoietin-2 and its receptor (Tie2) were also overexpressed (P < 0.05) in PTC. Correlations were found between ET-1 expression and that of NOS2, angiopoietin-1 and -2 (P < 0.05). NOS2 mRNA levels also correlated with those of NOS3 and angiopoietin-2 (P < 0.05). In thyroiditis, NOS2 immunoreactivity was observed in inflammatory cells whereas NOS2 mRNA levels were 12.1 +/- 1.6 times higher than normal (P < 0.005). CONCLUSIONS: This study revealed an activation of the NO pathway in thyroid carcinoma, which is interrelated to the ET-1 axis, both systems being overexpressed in concert with angiogenic factors. This global system might play a role in carcinogenesis and constitutes a potential target for anticancer therapy. PMID- 16712676 TI - Undetectable salivary testosterone in young women with premature ovarian failure. AB - BACKGROUND: The extent of androgen deficiency in young women with premature ovarian failure (POF) is unclear. AIM: Cross-sectional study of androgen status in young women with POF. PATIENTS: Twenty women with POF: six had Turner syndrome (group A); eight had iatrogenic POF either secondary to bilateral oophorectomy or treatment of malignancy (group B); and six had idiopathic POF (group C). The median age was 30.5 years (range 19-39); in groups B and C the median duration of ovarian failure was 10.0 years (range 1-35). METHODS: After a 2-month wash-out period without sex steroid replacement (SSR), serum testosterone (T), androstenedione (A4), dehydroepiandrosterone (DHEAS), SHBG, salivary testosterone (SalT) and the free androgen index [FAI = (serum T/SHBG) x 100] were measured. RESULTS: Median serum A4 was 4.6 nmol/l (10th, 90th centiles, 3.6, 5.1) and DHEAS was 3.2 micromol/l (10th, 90th centiles, 2.3, 9.3). Although median serum T was relatively low at 1.4 nmol/l (10th, 90th centiles, 1.1, 1.7), median SHBG was also low at 34 nmol/l (10th, 90th centiles 22.2, 67.5) and the median calculated FAI was within the normal range at 3.7 (10th, 90th centiles, 2.3, 7.0). However, SalT was undetectable in almost all subjects in the three groups of POF. CONCLUSIONS: Serum T and SHBG are relatively low in young women with POF and their FAI is therefore within the normal range. However, SalT, which measures free testosterone, is consistently low to undetectable in these young women with POF. The reliability of the FAI as a marker of androgen deficiency remains questionable. PMID- 16712677 TI - AntiMullerian serum levels during pregnancy. PMID- 16712678 TI - Association of HLA DQ4-DR8 haplotype with papillary thyroid carcinomas. PMID- 16712679 TI - Remission of a primary thyroid lymphoma after methotrexate withdrawal. PMID- 16712680 TI - Reversible dilated hypocalcaemic cardiomyopathy in a patient with primary hypoparathyroidism. PMID- 16712681 TI - Editing as a psychological practice. AB - The experience of the Jungian analyst in the role of editor of manuscripts by creative colleagues is examined. Historical precedents include Michael Fordham's editorial correspondence with Jung around the latter's synchronicity essay; Jung's handling of manuscripts submitted by Sabina Spielrein to the Jahrbuch fur psychoanalytische und psychopathologische Forschungen and various authors to the Zentralblatt fur Psychotherapie und ihre Grenzgebiete, and the author's close editing of a paper submitted by Andrew Samuels to the Journal of Analytical Psychology. In addition to mustering an adequate amount of generosity, erudition, and availability, the analytic editor must know how to clarify a psychological argument and to gauge the psychological impact of the written text. Notwithstanding transference/countertransference phenomena that can emerge around issues of competition, envy, and territoriality when author and editor are also fellow-authors working in the same field, the editor needs to be comfortable about serving as the author's selfobject and midwife. From an analytic perspective, although communicating decisions about the best way to put ideas into words can sometimes attract transference to the editor, the more profound transference that analysts experience in the editing situation is toward the text being edited, which helps to motivate donated time spent caring for journal manuscripts. PMID- 16712682 TI - Group process in Jungian analytic training and institute life. AB - Group process experience for analytic candidates is a neglected dimension of training, and receives little attention in the analytic literature. Jung observed group dynamics, but he never studied them closely, attending instead to the psychology of the individual. Unconscious currents in small groups have been studied by others, most notably by Wilfred Bion, and there are similarities between his theories of the group unconscious and Jung's theories of complexes. Experiential and didactic seminars in group process were added to the analytic curriculum at the C.G. Jung Institute of San Francisco in the early 1990s, leading to changes in the group dynamic of trainees and analysts alike. A discussion of the theories of Bion and Jung are followed by a report on our experiences of facilitating group process for analytic candidates. We give quotes from candidates and analyst members to illustrate the group process and its effects. The need for further study to develop a uniquely Jungian perspective on the unconscious structure and dynamics of the group is suggested. PMID- 16712684 TI - Queering gender: anima/animus and the paradigm of emergence. AB - An exploration into the world of the queer others of gender and sexuality moves us beyond the binary opposition of male/masculinity and female/femininity in our understanding of gender and expands the meaning of gender and sexuality for all humans. A revision of Jungian gender theory that embraces all genders and sexualities is needed not only to inform our clinical work but also to allow us to bring Jungian thought to contemporary gender theory and to cultural struggles such as gay marriage. The cognitive and developmental neurosciences are increasingly focused on the importance of body biology and embodied experience to the emergence of mind. In my exploration of gender I ask how gender comes to be experienced in a developing body and how those embodied gender feelings elaborate into a conscious category in the mind, a gender position. My understanding of emergent mind theory suggests that one's sense of gender, like other aspects of the mind, emerges very early in development from a self-organizing process involving an individual's particular body biology, the brain, and cultural environment. Gendered feeling, from this perspective, would be an emergent aspect of mind and not an archetypal inheritance, and the experiencing body would be key to gender emergence. A revised Jungian gender theory would transcend some of the limitations of Jung's anima/animus (A/A) gender thinking allowing us to contribute to contemporary gender theory in the spirit of another Jung; the Jung of the symbolic, the mythic, and the subtle body. This is the Jung who invites us to the medial place of the soul, bridging the realm of the physical body and the realm of the spirit. PMID- 16712683 TI - The analysis of the homoerotic and the pursuit of meaning. AB - This paper explores the dynamic tension between an evolving collective phenomenon and the nature of analytic process. Specifically, the focus will be erotic experiences which acquire a meaning through the culture at large, a meaning that may not be supportable when that material is subjected to psychological analysis. This stimulates a conflict between the symbolic attitude and the cultural perspective of the time. While the struggle between the individual and collective consciousness always emerges in analysis, the subject of same-gender sexual relations has become such a controversial and divisive issue in the current political environment that views toward homosexuality demand powerful allegiances and identification with either historic or contemporary ideas. People now identify as 'gay' and tend to see themselves as something akin to a race or perhaps alternative gender. Sexuality and relationship between same gendered people tends to be viewed through the lens of civil rights and the undeniable need for social equality. In this far-reaching and expanding collective phenomenon, psychology, in its support of human rights and accommodation to emerging trends, may be diminished in its capacity to pursue the meaning inherent in these human experiences. The position developed in this paper is that psychological experience, whether in the imaginal realm, dreams or personal consciousness, must be available for full analysis. Clinical experience and dreams are used to amplify this challenge to dynamic analytic practice. PMID- 16712685 TI - A word about words. AB - The title of my paper comes from an essay by Vaclav Havel. In his essay, Havel addressed most evocatively the question of the unique power of words for thinking and to influence, for good or for harm, as well as to inform and to educate. As analysts our medium is the word. I suggest that it is of inestimable importance that we are able to listen deeply to our patients' words and to be aware of our own. The quality of our ability to think deeply and consistently about the unconscious experience of our patient is intimately related to our ability to hear what is being said. The effect of our own words upon the patient likewise cannot be overstated. I offer my reflections on words in the analytic relationship and I give some clinical examples that I hope will illustrate my thoughts as to the power and the importance of what and how we hear and of what we say. PMID- 16712686 TI - Postmodern consciousness in psychotherapy. AB - Modern consciousness is a cultural and historical achievement in the West and a developmental task for each person now. Modern consciousness consists in the emancipation from the power of community, animistic nature and the unconscious. It is connected with neurosis and psychotherapy because it has to do with inner conflicts. But today there is an increasing number of cases which are characterized by dissociation and acting out, without the feeling of conflicts. Consciousness seems to be changing toward a new conception which might be called 'postmodern consciousness'. The essence of postmodern consciousness is shown by interpreting two dreams internally. The first dream from a case of depersonalization indicates that it is not necessary to be entangled with the object. There is a different kind of coniunctio in the mode of seeing. The second dream from a case of dissociative disorder shows a world which has neither traces of pre-modern cosmology-high and low, here and the beyond-nor modern interiority. There is only surface and self-reflection without content. The discussion of dreams suggests that postmodern consciousness is not to be understood as premature and pathological. It is therapeutically important to refine and deepen postmodern consciousness. PMID- 16712687 TI - Attachment-dissociation network: some thoughts about a modern complex theory. AB - The paper revises the complex theory in the light of modern infant research, neurosciences and object relation theory. The author takes up Jean Knox's idea to understand complexes as analogies to the internal working models of attachment theory. The author proposes to understand complexes as dissociated sub-networks out of the network structure of the psyche; these sub-networks contain the internal working models, the characteristic affects and unconscious expectation phantasies. With this network model one can try to understand severe defensive organizations in some patients as a pathological organization of different complexes. This is illustrated by a clinical example. PMID- 16712688 TI - Sexual metaphor, and the language of unconscious phantasy. PMID- 16712695 TI - Developmental malformations of the eye: the role of PAX6, SOX2 and OTX2. AB - Eye development initiates as an evagination of the early neural plate, before the closure of the neural tube. Structural malformations of the eye such as anophthalmia and microphthalmia arise very early in development. It is not surprising therefore that three of the genes currently identified to play a significant role in these developmental eye anomalies are also major players in brain development and regionalization. However, as has been emerging for a high proportion of transcriptional regulators studied, these genes have evolved to play multiple roles throughout development, and perhaps even in adult tissue maintenance. This complex spatiotemporal expression pattern requires elaborate regulatory systems which we are beginning to unravel. A major component of these complex regulatory networks is a series of cis-acting elements, highly conserved through evolution, which spread large distances from the coding region of each gene. We describe how cross regulation for PAX6, SOX2 and perhaps OTX2 has now been uncovered, pointing to the mechanisms that can fine-tune the expression of such essential developmental components. These interactions also help us understand why there is significant phenotypic overlap between mutations at these three loci. PMID- 16712696 TI - Craniofacial malformations: intrinsic vs extrinsic neural crest cell defects in Treacher Collins and 22q11 deletion syndromes. AB - The craniofacial complex is anatomically the most sophisticated part of the body. It houses all the major sensory organ systems and its origins are synonymous with vertebrate evolution. Of fundamental importance to craniofacial development is a specialized population of stem and progenitor cells, known as the neural crest, which generate the majority of the bone, cartilage, connective and peripheral nerve tissue in the head. Approximately one third of all congenital abnormalities exhibit craniofacial malformations and consequently, most craniofacial anomalies are considered to arise through primary defects in neural crest cell development. Recent advances however, have challenged this classical dogma, underscoring the influence of tissues with which the neural crest cells interact as the primary origin of patterning defects in craniofacial morphogenesis. In this review we discuss these neural crest cell interactions with mesoderm, endoderm and ectoderm in the head in the context of a better understanding of craniofacial malformations such as in Treacher Collins and 22q11 deletion syndromes. PMID- 16712700 TI - The personal experience of juvenile Huntington's disease: an interpretative phenomenological analysis of parents' accounts of the primary features of a rare genetic condition. AB - There has been a paucity of research into the psychosocial impact of juvenile Huntington's disease (JHD) on the child and the family. The study reported here is part of larger project that aimed to address this and investigate the social and health care needs of those affected by JHD. Ten semistructured interviews with the main caregiver(s) were carried out and were analyzed using the qualitative methodology interpretative phenomenological analysis. The main themes arising from the analysis are reported here: first becoming aware something is wrong; physical symptoms; speech and communication difficulties; behavioral problems; a slow but relentless process. These are discussed in relation to extant literature. We hope the article will be helpful to clinicians working with families where a child is affected by JHD and also contribute to the general literature on understanding symptoms in childhood illness. PMID- 16712701 TI - Factors in decision making following genetic counseling for pre-natal diagnosis of de novo chromosomal rearrangements. AB - A retrospective survey of genetic counselors was conducted in order to identify practice patterns and factors that influence a patient's decision making when a de novo translocation is diagnosed pre-natally. Different variables that influence patients' decisions about pregnancy management were assessed and compared. Specifically, the type of rearrangement and/or knowledge of the breakpoints, risks provided for abnormal outcome, anxiety, fetal ultrasound findings and personal reasons for parental decisions were evaluated. Our findings suggest that patient anxiety level significantly predicts pregnancy management decisions. This information may be of benefit in identifying potential areas of education for genetic counselors as well as other health care providers. PMID- 16712702 TI - High-efficiency multiplex capillary electrophoresis single strand conformation polymorphism (multi-CE-SSCP) mutation screening of SCN5A: a rapid genetic approach to cardiac arrhythmia. AB - Mutations in the SCN5A gene coding for the alpha-subunit of the cardiac Na(+) ion channel cause long QT syndrome, Brugada syndrome, idiopathic ventricular fibrillation, sick sinus node syndrome, progressive conduction disease, dilated cardiomyopathy and atrial standstill. These diseases exhibit variable expressivity, and identification of gene carriers is clinically important, particularly in sudden infant and adult death syndromes. The SCN5A gene comprises 28 exons distributed over 100 kbp of genomic sequence at chromosome 3p21. Disease causing mutations are private and scattered over the DNA sequence, making it difficult to screen for specific mutations. We developed a multiplex capillary electrophoresis single-strand conformation polymorphism (Multi-CE-SSCP) mutation screening protocol on the ABI 3100 platform and applied it to 10 previously slab gel SSCP identified mutations and SNPs and used it to identify one novel deletion. The method is highly efficient, with a turnover of 23 patients per 24 h and a false positive rate of 0.5% of the analyzed amplicons. Each variant has a particular elution pattern, and all 20 carriers of the H558R polymorphism out of 57 persons were correctly identified. We suggest that the method could become part of routine work-up of patients with suspicious syncope and of members of families with sudden unexplained death. PMID- 16712703 TI - Maternal MTR genotype contributes to the risk of non-syndromic cleft lip and palate in the Polish population. AB - The aetiology of non-syndromic cleft lip with or without cleft palate (CL/P) is very complex. It has been shown that polymorphic variants of genes encoding key proteins of folate and methionine metabolism might be important maternal risk factors of having a child with this craniofacial anomaly. Therefore, in our study, mothers with CL/P children as well as control mothers were examined for prevalence of polymorphisms of genes that encode methylenetetrahydrofolate reductase (MTHFR), methionine synthase (MTR), 5,10-methylenetetrahydrofolate dehydrogenase, 5,10-methenyltetrahydrofolate cyclohydrolase and 10 formyltetrahydrofolate synthetase (MTHFD1) and reduced folate carrier 1 (RFC1). We observed that there were no statistical differences in allele and genotype frequencies of MTHFR c.677C>T, MTHFD1 c.1958G>A and RFC1 c.80G>A between mothers who had children with CL/P and control mothers. However, mothers with MTR c.2756AG or GG genotype displayed a 2.195-fold increased risk of having a child with CL/P (95% CI 1.189-4.050, p = 0.011). The mechanism by which polymorphic transition of MTR gene might increase the maternal risk of having CL/P progeny is unknown. Our observations are consistent with a significant role of the methyl cycle in the development of craniofacial structures in humans. PMID- 16712704 TI - Genetic heterogeneity of synpolydactyly: a novel locus SPD3 maps to chromosome 14q11.2-q12. AB - Syndactyly type II or synpolydactyly (SPD) is the second most frequent syndactyly type and is inherited in an autosomal dominant fashion. The cardinal features of this malformation are the cutaneous or bony fusion of third and fourth fingers, and fourth and fifth toes associated with additional digital elements within the web. It shows incomplete penetrance and high inter- and intrafamilial phenotypic variability. Two loci are known for SPD (MIM 186000, MIM 608180) associated with mutations in HOXD13 and FBLN1, respectively. Here, we report further genetic heterogeneity for SDP. Employing a whole genomic screen, we demonstrate, in a large Pakistani kindred, that the classical phenotype of SPD maps on a new locus at chromosome 14q11.2-q12. The highest LOD score (Z(max) = 4.06) was obtained with microsatellite marker D14S264, and the multipoint LOD score reached a maximum of 5.01. Haplotype analysis revealed that the disease interval is flanked by microsatellite markers D14S283 and D14S1060, encompassing a physical distance of 10.72 Mb. We propose to allocate to this locus the symbol SPD3 (synpolydactyly 3), and to name the loci associated with HOXD13 or FBLN1 mutations SPD1 and SPD2, respectively. PMID- 16712705 TI - Crigler-Najjar type II syndrome may result from several types and combinations of mutations in the UGT1A1 gene. PMID- 16712706 TI - Multiple primary malignancies in a patient with situs ambiguus. PMID- 16712708 TI - Controversies in the public health approach to keratinocyte carcinomas. AB - Keratinocyte carcinomas are very common cancers in fair-skinned populations throughout the world. The term 'keratinocyte carcinoma' includes basal and squamous cell carcinoma of the skin, but not other cancers that may be included under the more ambiguous term 'nonmelanoma skin cancer'. Incidence and mortality are important for assessing impact and recent research suggests that these cancers are increasing in incidence in young adults. There is evidence to suggest that this trend may be due, in part, to the popularity of ultraviolet exposures in commercial tanning facilities. Mortality from keratinocyte carcinoma reveals distinct patterns suggestive of an important role of human papilloma virus infection. Ultimately, our aim is to reduce mortality and morbidity from these cancers, and to that end several large-scale trials of public health interventions have been conducted that may help point the way forward. PMID- 16712709 TI - Genetics and risk factors for basal cell carcinoma. AB - Nonmelanoma skin cancer (NMSC) is the commonest cancer in whites and its incidence is increasing worldwide. The prevalence of this cancer is predicted to equal that of all others combined and it was estimated that there were over 2 million cases diagnosed in the U.S.A. in 2004. Patients exhibit marked differences in clinical phenotype with variations in tumour numbers, rate of tumour accrual, site and histological subtype. Furthermore, patients are at increased risk of other cutaneous and noncutaneous cancers. The factors accounting for this variation are complex and still not completely understood. Clearly, ultraviolet light (UV) exposure is a major influence but its relationship to clinical phenotype is not yet clear. In addition, immunosuppression is a significant risk factor. Our group has identified high risk groups for the development of further basal cell carcinoma (BCC), namely patients with truncal BCC and those presenting with tumour clusters. This presentation will concentrate on these clinical subgroups as well as immunosuppressed patients. These groups represent significant management challenges and are areas where novel, nonsurgical treatment options may make a significant clinical impact in patient care. The risk factors predisposing to these clinical phenotypes will be discussed, including genetic factors and UV exposure. Potential clinical applications, including predictive indices, will be considered. PMID- 16712710 TI - Roles of the transcription factor p53 in keratinocyte carcinomas. AB - The transcription factor p53 is mutated in most keratinocyte carcinomas (nonmelanoma skin cancers). In these tumours, the gene bears the trace of its mutagen, sunlight. Sunlight-induced p53 mutations are also seen in skin precancers and even sun-exposed skin, which harbours thousands of p53-mutant keratinocyte clones. Normal p53 is upregulated by sunlight exposure, after which it acts as a tumour suppressor in several ways: increasing DNA repair, arresting the cell cycle and inducing apoptosis of badly damaged keratinocytes. This UV induced upregulation has been used as an assay for assessing the effectiveness of sunscreens. Once mutated, however, p53 renders cells apoptosis-resistant and therefore less sensitive to sunlight overexposure than normal cells. This reversal of roles drives clonal expansion of precancerous keratinocytes. PMID- 16712711 TI - Molecular mechanisms of cancer cell invasion and plasticity. AB - Cancer cell interactions with the extracellular matrix and the migration therein involve the function of adhesion receptors of the integrin family, a dynamic cytoskeleton, as well as proteolytic mechanisms to overcome tissue barriers. Recent progress in investigating tumour cell migration and associated matrix remodelling was made using three-dimensional (3D) dermis equivalents such as 3D collagen lattices or dermal explant cultures, prompting new concepts in molecular tumour invasion mechanisms and related adaptation responses. Mesenchymal HT-1080 fibrosarcoma cells as a model line migrate in an integrin-dependent manner and proteolytically cleave extracellular matrix structures. After interference with integrin and protease function, however, cancer cells can switch migration programs and thereby rescue migration by alternative mechanisms. Depending on the context of invasion, treatment with protease inhibitors or integrin antagonists can cause the mesenchymal-amoeboid transition and the collective-amoeboid transition, both generating sustained dissemination of single cells. These adaptation responses show an unexpected degree of plasticity resulting in migratory 'escape' strategies after pharmacotherapeutic intervention by prompting alternative mechanisms of cancer cell dissemination in tissues. PMID- 16712712 TI - Towards a more specific therapy: targeting nonmelanoma skin cancer cells. AB - Epithelial cancers of the skin, e.g. basal cell carcinoma and squamous cell carcinoma, are the most common tumours in humans with increasing incidence. Hence the development of new therapeutic strategies is of utmost interest. For many years the most often used conventional therapies for these diseases were surgical procedures such as curettage and electrodesiccation, excision or, with so far the best outcome in terms of remission rates, micrographic surgery. Other ablative treatment modalities are cryotherapy, radiation therapy or the use of lasers (Er:YAG, CO(2)). All those above-mentioned treatments have in common that they are quite unspecific and do not target the tumour itself or its environment, thus leading to unwanted effects in the surrounding tissue such as scar formation or other cosmetically disfiguring events. Therefore, the development of novel, more pathogenesis-based therapies such as the use of retinoids, cyclooxygenase inhibitors, topical immunomodulators, inhibitors of the sonic-hedgehog signalling pathway or photodynamic therapy are challenging new approaches. PMID- 16712715 TI - Differential cognitive impairment for diverse forms of multiple sclerosis. AB - BACKGROUND: Cognitive impairment is a common feature in multiple sclerosis (MS) patients and occurs in 60% of all cases. Unfortunately, neurological examination does not always agree with the neuropsychological evaluation in determining the cognitive profile of the patient. On the other hand, psychophysiological techniques such as event-related potentials (ERPs) can help in evaluating cognitive impairment in different pathologies. Behavioural responses and EEG signals were recorded during the experiment in three experimental groups: 1) a relapsing-remitting group (RRMS), 2) a benign multiple sclerosis group (BMS) and 3) a Control group. The paradigm employed was a spatial attention task with central cues (Posner experiment). The main aim was to observe the differences in the performance (behavioural variables) and in the latency and amplitude of the ERP components among these groups. RESULTS: Our data indicate that both MS groups showed poorer task performance (longer reaction times and lower percentage of correct responses), a latency delay for the N1 and P300 component, and a different amplitude for the frontal N1. Moreover, the deficit in the BMS group, indexed by behavioural and pyschophysiological variables, was more pronounced compared to the RRMS group. CONCLUSION: The present results suggest a cognitive impairment in the information processing in all of these patients. Comparing both pathological groups, cognitive impairment was more accentuated in the BMS group compared to the RMSS group. This suggests a silent deterioration of cognitive skills for the BMS that is not usually treated with pharmacological or neuropsychological therapy. PMID- 16712716 TI - The association of psychological stress and health related quality of life among patients with stroke and hypertension in Gaza Strip. AB - BACKGROUND: The study was performed to investigate the association of psychological stress and quality of life (QOL) among patients with the cardiovascular disease (CVD) of hypertension plus stroke or hypertension only. METHODS: The WHOQOL-BREF questionnaire was applied to 112 hypertensive patients with hypertension plus stroke and 224 hypertensive patients without stroke. Psychological stress was assessed with SCL-90. Means scale scores were compared using student-t-test and predictors of QOL were calculated with covariance analysis. RESULTS: Patients with stroke had a significant lower QOL than patients without stroke and a significantly higher level of stress (p < 0.01). In analyses of covariance psychological stress was significantly correlated to all domains of QOL among non-stroke patients. The same psychological and sociodemographic factors showed little impact on the stroke patients in these multivariable analyses. In these models psychological stress had a significant impact on the global domain of QOL among stroke patients. Income and gender were the only sociodemographic factors being significantly associated with the physical (education) and social (gender) domains of QOL in stroke patients. CONCLUSION: Psychological stress was strongly correlated with all domains of QOL in patients without stroke and was only partly associated with QOL among patients with stroke. Future studies should investigate if psychological stress is a factor suitable for educational and psychological interventions aiming at stress reduction in CVD patients which might substantially contribute to better health related quality of life in these patients. PMID- 16712717 TI - Attitudes and beliefs among patients treated with mood stabilizers. AB - BACKGROUND: There is increasing evidence that attitudes and beliefs are important in predicting adherence in depressive and bipolar disorders. However, such attitudes and beliefs on mood stabilizers have not been analysed by socio demographic and clinical variables. METHODS: The Mood Stabilizer Compliance Questionnaire (MSQC) was mailed to a large population of patients with depressive or bipolar disorder representative of patients treated at their first contacts to hospital settings in Denmark. RESULTS: Of the 1005 recipients, 49.9 % responded to the letter and among these 256 indicated that they previously had been or currently were in treatment with a mood stabilizer. A large proportion of the patients (40 to 80 %) had non-correct views on the effect of mood stabilizers. Older patients consistently had a more negative view on the doctor-patient relationship, more non-correct views on the effect of mood stabilizers and a more negative view on mood stabilizers. There was no difference in the attitudes and beliefs according to the type of disorder (depressive or bipolar), the number of psychiatric hospitalisations or according to the type of the current doctor (general practitioner, private psychiatrist, community psychiatry doctor, hospital doctor, other doctor). CONCLUSION: There is a need of improving knowledge and attitudes toward diagnosis and treatment especially among elder patients as this may add to improve the prognosis of depressive and bipolar disorders. PMID- 16712713 TI - Anti-inflammatory and immunosuppressive drugs and reproduction. AB - Rheumatic diseases in women of childbearing years may necessitate drug treatment during a pregnancy, to control maternal disease activity and to ensure a successful pregnancy outcome. This survey is based on a consensus workshop of international experts discussing effects of anti-inflammatory, immunosuppressive and biological drugs during pregnancy and lactation. In addition, effects of these drugs on male and female fertility and possible long-term effects on infants exposed to drugs antenatally are discussed where data were available. Recommendations for drug treatment during pregnancy and lactation are given. PMID- 16712718 TI - A paradox of immunodeficiency and inflammation in human aging: lessons learned from apoptosis. AB - Aging is associated with a paradox of immunodeficiency and inflammation (an evidence of hyperactive immune system). Apoptosis is associated with cellular depletion and suppression of inflammatory response. In this brief review, we will present evidence for the role of increased apoptosis in immunodeficiency and paradoxical increased inflammation associated with human aging. In particular, a role of apoptotic cells in failure to generate anti-inflammatory responses and directly activating inflammatory responses will be discussed. PMID- 16712720 TI - Low serum albumin and the acute phase response predict low serum selenium in HIV 1 infected women. AB - BACKGROUND: Low serum selenium has been associated with lower CD4 counts and greater mortality among HIV-1-seropositive individuals, but most studies have not controlled for serum albumin and the presence of an acute phase response. METHODS: A cross-sectional study was conducted to evaluate relationships between serum selenium concentrations and CD4 count, plasma viral load, serum albumin, and acute phase response markers among 400 HIV-1-seropositive women. RESULTS: In univariate analyses, lower CD4 count, higher plasma viral load, lower albumin, and the presence of an acute phase response were each significantly associated with lower serum selenium concentrations. In multivariate analyses including all four of these covariates, only albumin remained significantly associated with serum selenium. For each 0.1 g/dl increase in serum albumin, serum selenium increased by 0.8 microg/l (p < 0.001). Women with an acute phase response also had lower serum selenium (by 5.6 microg/l, p = 0.06). CONCLUSION: Serum selenium was independently associated with serum albumin, but not with CD4 count or plasma viral load, in HIV-1-seropositive women. Our findings suggest that associations between lower serum selenium, lower CD4 count, and higher plasma viral load may be related to the frequent occurrence of low serum albumin and the acute phase response among individuals with more advanced HIV-1 infection. PMID- 16712721 TI - A nucleolar localizing Rev binding element inhibits HIV replication. AB - The Rev protein of the human immunodeficiency virus (HIV) facilitates the nuclear export of intron containing viral mRNAs allowing formation of infectious virions. Rev traffics through the nucleolus and shuttles between the nucleus and cytoplasm. Rev multimerization and interaction with the export protein CRM1 takes place in the nucleolus. To test the importance of Rev nucleolar trafficking in the HIV-1 replication cycle, we created a nucleolar localizing Rev Response Element (RRE) decoy and tested this for its anti-HIV activity. The RRE decoy provided marked inhibition of HIV-1 replication in both the CEM T-cell line and in primary CD34+ derived monocytes. These results demonstrate that titration of Rev in the nucleolus impairs HIV-1 replication and supports a functional role for Rev trafficking in this sub-cellular compartment. PMID- 16712719 TI - HIV-1 associated dementia: symptoms and causes. AB - Despite the use of highly active antiretroviral therapy (HAART), neuronal cell death remains a problem that is frequently found in the brains of HIV-1-infected patients. HAART has successfully prevented many of the former end-stage complications of AIDS, however, with increased survival times, the prevalence of minor HIV-1 associated cognitive impairment appears to be rising among AIDS patients. Further, HIV-1 associated dementia (HAD) is still prevalent in treated patients as well as attenuated forms of HAD and CNS opportunistic disorders. HIV associated cognitive impairment correlates with the increased presence in the CNS of activated, though not necessarily HIV-1-infected, microglia and CNS macrophages. This suggests that indirect mechanisms of neuronal injury and loss/death occur in HIV/AIDS as a basis for dementia since neurons are not themselves productively infected by HIV-1. In this review, we discussed the symptoms and causes leading to HAD. Outcome from this review will provide new information regarding mechanisms of neuronal loss in AIDS patients. PMID- 16712722 TI - Globalization and local response to epidemiological overlap in 21st century Ecuador. AB - BACKGROUND: Third World countries are confronted by a complex overlay of two sets of health problems. Traditional maladies, including communicable diseases, malnutrition, and environmental health hazards coexist with emerging health challenges, including cardiovascular disease, cancer, and increasing levels of obesity. Using Ecuador as an example, this paper proposes a conceptual framework for linking epidemiologic overlap to emerging social structures and processes at the national and global levels. DISCUSSION: Epidemiologic trends can be seen as part of broader processes related to globalization, but this does not imply that globalization is a monolithic force that inevitably and uniformly affects nations, communities, and households in the same manner. Rather, characteristics and forms of social organization at the subnational level can shape the way that globalization takes place. Thus, globalization has affected Ecuador in specific ways and is, at the same time, intimately related to the form in which the epidemiologic transition has transpired in that country. SUMMARY: Ecuador is among neither the poorest nor the wealthiest countries and its situation may illuminate trends in other parts of the world. As in other countries, insertion into the global economy has not taken place in a vacuum; rather, Ecuador has experienced unprecedented social and demographic change in the past several decades, producing profound transformation in its social structure. Examples of local represent alternatives to centralized health systems that do not effectively address the complex overlay of traditional and emerging health problems. PMID- 16712723 TI - Adrenalectomy promotes a permanent decrease of plasma corticoid levels and a transient increase of apoptosis and the expression of Transforming Growth Factor beta1 (TGF-beta1) in hippocampus: effect of a TGF-beta1 oligo-antisense. AB - BACKGROUND: Corticosterone reduction produced by adrenalectomy (ADX) induces apoptosis in dentate gyrus (DG) of the hippocampus, an effect related to an increase in the expression of the pro-apoptotic gene bax. However it has been reported that there is also an increase of the anti-apoptotic gene bcl-2, suggesting the promotion of a neuroprotective phenomenon, perhaps related to the expression of transforming growth factor beta1 (TGF-beta1). Thus, we have investigated whether TGF-beta1 levels are induced by ADX, and whether apoptosis is increased by blocking the expression of TGF-beta1 with an antisense oligonucleotide (ASO) administered intracerebrally in corticosterone depleted rats. RESULTS: It was observed an increase of apoptosis in DG, 2 and 5 days after ADX, in agreement with a reduction of corticosterone levels. However, the effect of ADX on the number of apoptotic positive cells in DG was decreased 5 days after the lesion. In CA1-CA3 regions, the effect was only observed 2 days after ADX. TGF-beta1 mRNA levels were increased 2 days after ADX. The sustained intracerebro ventricular administration of a TGF-beta1 ASO via an osmotic mini pump increased apoptosis levels in CA and DG regions 5 days after ADX as well as sham-operated control animals. No significant effect was observed following a scrambled oligodeoxynucleotide treatment. CONCLUSION: The changes in both the pattern and the magnitude of apoptotic-cell morphology observed 2 and 5 days after ADX suggest that, as a consequence of the reduction of corticosteroids, some trophic mechanisms restricting cell death to a particular time window are elicited. Sustained intracerebral administration of TGF-beta1 ASO increased the apoptosis promoted by ADX, suggesting that TGF-beta1 plays an anti-apoptotic role in vivo in hippocampus. PMID- 16712724 TI - Slow fertilization of stickleback eggs: the result of sexual conflict? AB - BACKGROUND: The fertilization success in sperm competition in externally fertilizing fish depends on number and quality of sperm. The time delay between sequential ejaculations may further influence the outcome of sperm competition. Such a time interval can load the raffle over fertilization if fertilization takes place very fast. Short fertilization times are generally assumed for externally fertilizing fish such as the three-spined stickleback (Gasterosteus aculeatus). In this pair-spawning fish, territorial males often try to steal fertilizations in nests of neighbouring males. This sneaking behaviour causes sperm competition. Sneakers will only get a share of paternity when eggs are not fertilized immediately after sperm release. Contrary to males, females may be interested in multiple paternity of their clutch of eggs. There thus may be a sexual conflict over the speed of fertilization. RESULTS: In this study we used two different in vitro fertilization experiments to assess how fast eggs are fertilized in sticklebacks. We show that complete fertilization takes more than 5 min which is atypically long for externally fertilizing fishes. CONCLUSION: This result suggests that the time difference does not imply high costs to the second stickleback male to ejaculate. Slow fertilization (and concomitant prolonged longevity of sperm) may be the result of sexual conflict in which females aimed at complete fertilization and/or multiple paternity. PMID- 16712726 TI - On the persistence of supplementary resources in biomedical publications. AB - BACKGROUND: Providing for long-term and consistent public access to scientific data is a growing concern in biomedical research. One aspect of this problem can be demonstrated by evaluating the persistence of supplementary data associated with published biomedical papers. METHODS: We manually evaluated 655 supplementary data links extracted from PubMed abstracts published 1998-2005 (Method 1) as well as a further focused subset of 162 full-text manuscripts published within three representative high-impact biomedical journals between September and December 2004 (Method 2). RESULTS: For Method 1 we found that since 2001, only 71 - 92% of supplementary data were still accessible via the links provided, with 93% of these inaccessible links occurring where supplementary data was not stored with the publishing journal. Of the manuscripts evaluated in Method 2, we found that only 83% of these links were available approximately a year after publication, with 55% of these inaccessible links were at locations outside the journal of publication. CONCLUSION: We conclude that if supplemental data is required to support the publication, journals policies must take-on the responsibility to accept and store such data or require that it be maintained with a credible independent institution or under the terms of a strategic data storage plan specified by the authors. We further recommend that publishers provide automated systems to ensure that supplementary links remain persistent, and that granting bodies such as the NIH develop policies and funding mechanisms to maintain long-term persistent access to these data. PMID- 16712727 TI - Empirical array quality weights in the analysis of microarray data. AB - BACKGROUND: Assessment of array quality is an essential step in the analysis of data from microarray experiments. Once detected, less reliable arrays are typically excluded or "filtered" from further analysis to avoid misleading results. RESULTS: In this article, a graduated approach to array quality is considered based on empirical reproducibility of the gene expression measures from replicate arrays. Weights are assigned to each microarray by fitting a heteroscedastic linear model with shared array variance terms. A novel gene-by gene update algorithm is used to efficiently estimate the array variances. The inverse variances are used as weights in the linear model analysis to identify differentially expressed genes. The method successfully assigns lower weights to less reproducible arrays from different experiments. Down-weighting the observations from suspect arrays increases the power to detect differential expression. In smaller experiments, this approach outperforms the usual method of filtering the data. The method is available in the limma software package which is implemented in the R software environment. CONCLUSION: This method complements existing normalisation and spot quality procedures, and allows poorer quality arrays, which would otherwise be discarded, to be included in an analysis. It is applicable to microarray data from experiments with some level of replication. PMID- 16712725 TI - Differential gene expression in abdomens of the malaria vector mosquito, Anopheles gambiae, after sugar feeding, blood feeding and Plasmodium berghei infection. AB - BACKGROUND: Large scale sequencing of cDNA libraries can provide profiles of genes expressed in an organism under defined biological and environmental circumstances. We have analyzed sequences of 4541 Expressed Sequence Tags (ESTs) from 3 different cDNA libraries created from abdomens from Plasmodium infection susceptible adult female Anopheles gambiae. These libraries were made from sugar fed (S), rat blood fed (RB), and P. berghei-infected (IRB) mosquitoes at 30 hours after the blood meal, when most parasites would be transforming ookinetes or very early oocysts. RESULTS: The S, RB and IRB libraries contained 1727, 1145 and 1669 high quality ESTs, respectively, averaging 455 nucleotides (nt) in length. They assembled into 1975 consensus sequences--567 contigs and 1408 singletons. Functional annotation was performed to annotate probable molecular functions of the gene products and the biological processes in which they function. Genes represented at high frequency in one or more of the libraries were subjected to digital Northern analysis and results on expression of 5 verified by qRT-PCR. CONCLUSION: 13% of the 1965 ESTs showing identity to the A. gambiae genome sequence represent novel genes. These, together with untranslated regions (UTR) present on many of the ESTs, will inform further genome annotation. We have identified 23 genes encoding products likely to be involved in regulating the cellular oxidative environment and 25 insect immunity genes. We also identified 25 genes as being up or down regulated following blood feeding and/or feeding with P. berghei infected blood relative to their expression levels in sugar fed females. PMID- 16712728 TI - Analysis of proliferative activity in oral gingival epithelium in immunosuppressive medication induced gingival overgrowth. AB - BACKGROUND: Drug-induced gingival overgrowth is a frequent adverse effect associated principally with administration of the immunosuppressive drug cyclosporin A and also certain antiepileptic and antihypertensive drugs. It is characterized by a marked increase in the thickness of the epithelial layer and accumulation of excessive amounts of connective tissue. The mechanism by which the drugs cause gingival overgrowth is not yet understood. The purpose of this study was to compare proliferative activity of normal human gingiva and in cyclosporine A-induced gingival overgrowth. METHODS: Gingival samples were collected from 12 generally healthy individuals and 22 Cyclosporin A-medicated renal transplant recipients. Expression of proliferating cell nuclear antigen was evaluated in formalin-fixed, paraffin-embedded gingival samples using an immunoperoxidase technique and a monoclonal antibody for this antigen. RESULTS: There were differences between the Cyclosporin A group and control group in regard to proliferating cell nuclear antigen and epithelial thickness. In addition, the degree of stromal inflammation was higher in the Cyclosporin A group when compared with the control group. CONCLUSION: The results suggest that the increased epithelial thickness observed in Cyclosporin A-induced gingival overgrowth is associated with increased proliferative activity in keratinocytes. PMID- 16712729 TI - Fluid structure interaction of patient specific abdominal aortic aneurysms: a comparison with solid stress models. AB - BACKGROUND: Abdominal aortic aneurysm (AAA) is a dilatation of the aortic wall, which can rupture, if left untreated. Previous work has shown that, maximum diameter is not a reliable determinant of AAA rupture. However, it is currently the most widely accepted indicator. Wall stress may be a better indicator and promising patient specific results from structural models using static pressure, have been published. Since flow and pressure inside AAA are non-uniform, the dynamic interaction between the pulsatile flow and wall may influence the predicted wall stress. The purpose of the present study was to compare static and dynamic wall stress analysis of patient specific AAAs. METHOD: Patient-specific AAA models were created from CT scans of three patients. Two simulations were performed on each lumen model, fluid structure interaction (FSI) model and static structural (SS) model. The AAA wall was created by dilating the lumen with a uniform 1.5 mm thickness, and was modeled as a non-linear hyperelastic material. Commercial finite element code Adina 8.2 was used for all simulations. The results were compared between the FSI and SS simulations. RESULTS: Results are presented for the wall stress patterns, wall shear stress patterns, pressure, and velocity fields within the lumen. It is demonstrated that including fluid flow can change local wall stresses slightly. However, as far as the peak wall stress is concerned, this effect is negligible as the difference between SS and FSI models is less than 1%. CONCLUSION: The results suggest that fully coupled FSI simulation, which requires considerable computational power to run, adds little to rupture risk prediction. This justifies the use of SS models in previous studies. PMID- 16712730 TI - Splenectomy and proximal lieno-renal shunt in a factor five deficient patient with extra-hepatic portal vein obstruction. AB - BACKGROUND: The clinico-surgical implication and successful management of a rare case of factor five (V) deficiency with portal hypertension and hypersplenism due to idiopathic extra-hepatic portal venous obstruction is presented. CASE PRESENTATION: A 16-year old boy had gastro-esophageal variceal bleeding, splenomegaly and hypersplenism. During preoperative workup prolonged prothrombin time and activated partial thromboplastin time were detected, which on further evaluation turned out to be due to factor V deficiency. Proximal lieno-renal shunt and splenectomy were successfully performed with transfusion of fresh frozen plasma during and after the surgical procedure. At surgery there was no excessive bleeding. The perioperative course was uneventful and the patient is doing well on follow up. CONCLUSION: Surgical portal decompressive procedures can be safely undertaken in clotting factor deficient patients with portal hypertension if meticulous surgical hemostasis is achieved at operation and the deficient factor is adequately replaced in the perioperative period. PMID- 16712731 TI - The impact of clothing style on bone mineral density among post menopausal women in Morocco: a case-control study. AB - BACKGROUND: The clothing style is an important factor that influences vitamin D production and thus bone mineral density. We performed a case-control study in order to evaluate the effect of veil wearing (concealing clothing) on bone mineral density in Moroccan post menopausal women. METHODS: The cases were osteoporotic women whose disease was assessed by bone mineral density measurement. Each patient was matched with a non osteoporotic woman for age, and body mass index. All our patients were without secondary causes or medications that might affect bone density. The veil was defined as a concealing clothing which covered most of the body including the arms, the legs and the head. This definition is this of the usual Moroccan traditional clothing style. RESULTS: 178 post menopausal osteoporotic patients and 178 controls were studied. The mean age of the cases and the controls was 63.2 years (SD 7) and the mean body mass index was 32.1 (SD 8). The results of crude Odds Ratios analyses indicated that wearing a veil was associated with a high risk of osteoporosis: OR 2.29 (95% CI, 1.38 3.82). Multiparity or a history of familial peripheral osteoporotic fractures had also a significant effect on increasing the osteoporosis risk (ORs: 1.87 (95% CI, 1.05-3.49) and 2.01 (95% CI, 1.20-3.38)). After a multiple regression analysis, wearing the veil and a history of familial osteoporotic fractures remained the both independent factors that increased the osteoporosis risk (ORs: 2.20 (95% CI, 1.22-3.9) and 2.19 (95% CI, 1.12-4.29) respectively). CONCLUSION: our study suggested that in Moroccan post menopausal women, wearing a traditional concealing clothing covering arms, legs and head increased the risk of osteoporosis. Further studies are required to evaluate the clinical impact of the above findings and to clarify the status of vitamin D among veiled women in Morocco. PMID- 16712732 TI - Predicting DNA-binding sites of proteins from amino acid sequence. AB - BACKGROUND: Understanding the molecular details of protein-DNA interactions is critical for deciphering the mechanisms of gene regulation. We present a machine learning approach for the identification of amino acid residues involved in protein-DNA interactions. RESULTS: We start with a Naive Bayes classifier trained to predict whether a given amino acid residue is a DNA-binding residue based on its identity and the identities of its sequence neighbors. The input to the classifier consists of the identities of the target residue and 4 sequence neighbors on each side of the target residue. The classifier is trained and evaluated (using leave-one-out cross-validation) on a non-redundant set of 171 proteins. Our results indicate the feasibility of identifying interface residues based on local sequence information. The classifier achieves 71% overall accuracy with a correlation coefficient of 0.24, 35% specificity and 53% sensitivity in identifying interface residues as evaluated by leave-one-out cross-validation. We show that the performance of the classifier is improved by using sequence entropy of the target residue (the entropy of the corresponding column in multiple alignment obtained by aligning the target sequence with its sequence homologs) as additional input. The classifier achieves 78% overall accuracy with a correlation coefficient of 0.28, 44% specificity and 41% sensitivity in identifying interface residues. Examination of the predictions in the context of 3-dimensional structures of proteins demonstrates the effectiveness of this method in identifying DNA-binding sites from sequence information. In 33% (56 out of 171) of the proteins, the classifier identifies the interaction sites by correctly recognizing at least half of the interface residues. In 87% (149 out of 171) of the proteins, the classifier correctly identifies at least 20% of the interface residues. This suggests the possibility of using such classifiers to identify potential DNA-binding motifs and to gain potentially useful insights into sequence correlates of protein-DNA interactions. CONCLUSION: Naive Bayes classifiers trained to identify DNA-binding residues using sequence information offer a computationally efficient approach to identifying putative DNA-binding sites in DNA-binding proteins and recognizing potential DNA-binding motifs. PMID- 16712733 TI - Effect of 5'UTR introns on gene expression in Arabidopsis thaliana. AB - BACKGROUND: The majority of introns in gene transcripts are found within the coding sequences (CDSs). A small but significant fraction of introns are also found to reside within the untranslated regions (5'UTRs and 3'UTRs) of expressed sequences. Alignment of the whole genome and expressed sequence tags (ESTs) of the model plant Arabidopsis thaliana has identified introns residing in both coding and non-coding regions of the genome. RESULTS: A bioinformatic analysis revealed some interesting observations: (1) the density of introns in 5'UTRs is similar to that in CDSs but much higher than that in 3'UTRs; (2) the 5'UTR introns are preferentially located close to the initiating ATG codon; (3) introns in the 5'UTRs are, on average, longer than introns in the CDSs and 3'UTRs; and (4) 5'UTR introns have a different nucleotide composition to that of CDS and 3'UTR introns. Furthermore, we show that the 5'UTR intron of the A. thaliana EF1alpha-A3 gene affects the gene expression and the size of the 5'UTR intron influences the level of gene expression. CONCLUSION: Introns within the 5'UTR show specific features that distinguish them from introns that reside within the coding sequence and the 3'UTR. In the EF1alpha-A3 gene, the presence of a long intron in the 5'UTR is sufficient to enhance gene expression in plants in a size dependent manner. PMID- 16712734 TI - Prognostic significance of cyclooxygenase-2 (COX-2) expression in patients with surgically resectable adenocarcinoma of the oesophagus. AB - BACKGROUND: COX-2 expression in tumour cells has been associated with poor prognosis in gastrointestinal and non-gastrointestinal cancers. The aim of our study was to test the hypothesis that higher levels of COX-2 expression are prognostically related to poor clinico-pathologic features in adenocarcinoma of the oesophagus. METHODS: We reviewed the records of 100 consecutive patients undergoing resection for adenocarcinoma of the oesophagus to collect data on T stage, N-stage, tumour recurrence and survival. T & N-stage was further confirmed by histological examination. COX-2 protein expression was assessed by immunohistochemistry in all patients and COX-2 m-RNA expression was measured by quantitative RT-PCR in a small group of patients. RESULTS: Higher levels of COX-2 expression were associated with higher T stage (p = 0.008), higher N stage (p = 0.049), increased risk of tumour recurrence (p = 0.01) and poor survival (p = <0.001). A COX-2 score of >200 was associated with a median survival of 10 months compared to 26 months with a score of <200 (p = <0.001). CONCLUSION: Higher levels of COX-2 expression are associated with poor clinico-pathologic features and poor survival in patients with oesophageal adenocarcinoma. PMID- 16712735 TI - Core set approach to reduce uncertainty of gene trees. AB - BACKGROUND: A genealogy based on gene sequences within a species plays an essential role in the estimation of the character, structure, and evolutionary history of that species. Because intraspecific sequences are more closely related than interspecific ones, detailed information on the evolutionary process may be available by determining all the node sequences of trees and provide insight into functional constraints and adaptations. However, strong evolutionary correlations on a few lineages make this determination difficult as a whole, and the maximum parsimony (MP) method frequently allows a number of topologies with a same total branching length. RESULTS: Kitazoe et al. developed multidimensional vector-space representation of phylogeny. It converts additivity of evolutionary distances to orthogonality among the vectors expressing branches, and provides a unified index to measure deviations from the orthogoality. In this paper, this index is used to detect and exclude sequences with large deviations from orthogonality, and then selects a maximum subset ("core set") of sequences for which MP generates a single solution. Once the core set tree is formed whose all the node sequences are given, the excluded sequences are found to have basically two phylogenetic positions on this tree, respectively. Fortunately, since multiple substitutions are rare in intra-species sequences, the variance of nucleotide transitions is confined to a small range. By applying the core set approach to 38 partial env sequences of HIV-1 in a single patient and also 198 mitochondrial COI and COII DNA sequences of Anopheles dirus, we demonstrate how consistently this approach constructs the tree. CONCLUSION: In the HIV dataset, we confirmed that the obtained core set tree is the unique maximum set for which MP proposes a single tree. In the mosquito data set, the fluctuation of nucleotide transitions caused by the sequences excluded from the core set was very small. We reproduced this core-set tree by simulation based on random process, and applied our approach to many sets of the obtained endpoint sequences. Consequently, the ninety percent of the endpoint sequences was identified as the core sets and the obtained node sequences were perfectly identical to the true ones. PMID- 16712736 TI - Long-term clearance from small airways in subjects with ciliary dysfunction. AB - The objective of this study was to investigate if long-term clearance from small airways is dependent on normal ciliary function. Six young adults with primary ciliary dyskinesia (PCD) inhaled 111 Indium labelled Teflon particles of 4.2 microm geometric and 6.2 microm aerodynamic diameter with an extremely slow inhalation flow, 0.05 L/s. The inhalation method deposits particles mainly in the small conducting airways. Lung retention was measured immediately after inhalation and at four occasions up to 21 days after inhalation. Results were compared with data from ten healthy controls. For additional comparison three of the PCD subjects also inhaled the test particles with normal inhalation flow, 0.5 L/s, providing a more central deposition. The lung retention at 24 h in % of lung deposition (Ret24) was higher (p < 0.001) in the PCD subjects, 79 % (95% Confidence Interval, 67.6;90.6), compared to 49% (42.3;55.5) in the healthy controls. There was a significant clearance after 24 h both in the PCD subjects and in the healthy controls with equivalent clearance. The mean Ret24 with slow inhalation flow was 73.9 +/- 1.9% compared to 68.9 +/- 7.5% with normal inhalation flow in the three PCD subjects exposed twice. During day 7-21 the three PCD subjects exposed twice cleared 9% with normal flow, probably representing predominantly alveolar clearance, compared to 19% with slow inhalation flow, probably representing mainly small airway clearance. This study shows that despite ciliary dysfunction, clearance continues in the small airways beyond 24 h. There are apparently additional clearance mechanisms present in the small airways. PMID- 16712737 TI - A simple and rapid method for detection of Trypanosoma evansi in the dromedary camel using a nested polymerase chain reaction. AB - A nested polymerase chain reaction (nPCR)-based assay, was developed and evaluated for rapid detection of Trypanosoma evansi in experimentally infected mice and naturally infected camels (Camelus dromedarius). Four oligonucleotide primers (TE1, TE2, TE3 and TE4), selected from nuclear repetitive gene of T. evansi, were designed and used for PCR amplifications. The first amplification, using a pair of outer primers TE1 and TE2, produced a 821-bp primary PCR product from T. evansi DNA. The second amplification, using nested (internal) pair of primers TE3 and TE4, produced a 270-bp PCR product. T. evansi DNAs extracted from blood samples of experimentally infected mice and naturally infected Sudanese breed of dromedary camels were detected by this nested PCR-based assay. The nested primers TE3 and TE4 increased the sensitivity of the PCR assay and as little as 10 fg of T. evansi DNA (equivalent to a single copy of the putative gene of the parasite) was amplified and visualized onto ethidium bromide-stained agarose gels. Amplification products were not detected when the PCR-based assay was applied to DNA from other blood parasites including Thieleria annulata, Babesia bigemina or nucleic acid free samples. Application of this nPCR-based assay to clinical samples resulted in direct detection of T. evansi from a variety of tissue samples collected from experimentally infected mice and blood from naturally infected camels. The described nPCR-based assay provides a valuable tool to study the epidemiology of T. evansi infection in camels and other susceptible animal populations. PMID- 16712738 TI - Comparison of coverage with insecticide-treated nets in a Tanzanian town and villages where nets and insecticide are either marketed or provided free of charge. AB - BACKGROUND: There is much emphasis on social marketing as a means of scaling up coverage with insecticide-treated nets and the question has arisen whether nets provided free-of-charge will be looked after by householders. METHODS: Over several years questionnaires and surveys of usage and condition of nets were carried out throughout a town and 15 villages in north-east Tanzania, where nets and insecticide have to be purchased and in 24 other villages where over 15000 nets had been donated and annual re-treatment is provided free-of-charge. RESULTS: There was very high population coverage in the town but, in the villages where nets have to be purchased, only 9.3% of people used nets which were intact and/or had been insecticide-treated and could, therefore, provide protection. However, where nets had been provided free, over 90% of the nets were still present and were brought for re-treatment several years later. CONCLUSION: In this part of Tanzania, social marketing has performed well in a town but very poorly in villages. However, the study showed that people look after and bring for re-treatment nets which had been provided free-of-charge. PMID- 16712739 TI - Automatic generation of gene finders for eukaryotic species. AB - BACKGROUND: The number of sequenced eukaryotic genomes is rapidly increasing. This means that over time it will be hard to keep supplying customised gene finders for each genome. This calls for procedures to automatically generate species-specific gene finders and to re-train them as the quantity and quality of reliable gene annotation grows. RESULTS: We present a procedure, Agene, that automatically generates a species-specific gene predictor from a set of reliable mRNA sequences and a genome. We apply a Hidden Markov model (HMM) that implements explicit length distribution modelling for all gene structure blocks using acyclic discrete phase type distributions. The state structure of the each HMM is generated dynamically from an array of sub-models to include only gene features represented in the training set. CONCLUSION: Acyclic discrete phase type distributions are well suited to model sequence length distributions. The performance of each individual gene predictor on each individual genome is comparable to the best of the manually optimised species-specific gene finders. It is shown that species-specific gene finders are superior to gene finders trained on other species. PMID- 16712740 TI - [Relationship between mortality due to acute myocardial infarction and adherence to clinical guidelines]. PMID- 16712741 TI - [Cardiac transplantation in Spain. Have we reached our peak?]. PMID- 16712742 TI - [Myocardial disease as a cause of sudden cardiac death syndrome in children and young adults: recognition and preventive strategies needed]. PMID- 16712743 TI - [Reduction in acute myocardial infarction mortality over a five-year period]. AB - INTRODUCTION AND OBJECTIVES: To assess recent changes in the management of patients with acute myocardial infarction (AMI) and their impact on mortality using data from the PRIAMHO I and II registries (1995 and 2000). PATIENTS AND METHOD: Of the 168 public hospitals in Spain, 24 and 58 contributed to the 1995 and 2000 PRIAMHO registries, respectively. RESULTS: Patients in the PRIAMHO II registry (n=6221) were significantly older, more often female, and proportionally more likely to have coronary risk factors or a previous myocardial infarction, or to have undergone revascularization than those in PRIAMHO I (n=5242). Reperfusion therapy was administered more often (46.9% vs 41.9%, P<.001) and earlier (48 min vs 60 min, P<.001). Antiplatelet drugs were given to 96.1% vs 89.1% of patients, beta-blockers to 51.1% vs 30.1%, and ACE inhibitors to 41.6% vs 24.9% (P<.001 for all comparisons). In addition, 28-day mortality was 11.3% and 14.2% (P<.001), respectively, and one-year mortality, 16.4% and 18.5% (P<.001), respectively. The adjusted hazard ratio for mortality at one year in PRIAMHO II compared with PRIAMHO I was 0.78 (95% CI, 0.70-0.86, P<.001; adjusted for age, sex, diabetes, smoking, dyslipemia, hypertension, previous MI and CABG, ST-elevation status and Killip class at admission, and hospital characteristics). CONCLUSIONS: Even though patients registered in 2000 formed a higher risk group than those registered in 1995, one-year mortality after AMI decreased by 22% over the five year period. This improvement was due to more frequent and earlier reperfusion therapy and better use of antithrombotics, beta-blockers and ACE inhibitors. PMID- 16712744 TI - [Prognostic value of serum creatinine in non-ST-elevation acute coronary syndrome]. AB - INTRODUCTION AND OBJECTIVES: Cardiovascular disease is the main cause of death in patients with kidney failure. Moreover, the presence of impaired renal function is an important prognostic factor in patients with heart disease, and is a determinant of outcome during follow-up. The main aim was to investigate the relationship between kidney failure at admission and one-year mortality in patients with non-ST-elevation acute coronary syndrome. PATIENTS AND METHOD: We studied 1029 consecutive patients admitted to our institution. The serum creatinine level and glomerular filtration rate were determined at admission, and classical risk factors and biochemical markers were assessed. The primary endpoint was all-cause mortality at one year. RESULTS: Patients who died were older, more frequently had a history of diabetes or coronary artery disease, were more likely to have heart failure at admission, had higher troponin-I, myoglobin and creatinine levels, and were less likely to have dyslipidemia or to be a smoker. Multivariate analysis showed that the independent predictors of all-cause mortality at one year were age, diabetes, troponin-I level, Killip class > 1, male gender, creatinine level, and glomerular filtration rate. There was a linear correlation between increased risk and creatinine level. CONCLUSIONS: Creatinine level at admission is one of the most important covariates in early prognostic stratification in these patients. A high serum creatinine level (or a low glomerular filtration rate) increases the probability of death due to all causes. The serum creatinine level is, moreover, an inexpensive, easy-to-use, and widely available prognostic marker. PMID- 16712745 TI - [Direct stent implantation in acute myocardial infarction. The DISCO 3 study]. AB - INTRODUCTION AND OBJECTIVES: An association has been reported between direct stenting in primary angioplasty and low incidences of the no-reflow phenomenon and distal embolization. The aims of this study were to determine the proportion of patients who can be treated by direct stent implantation and to identify factors that establish when the technique should be used in acute myocardial infarction in clinical practice. PATIENTS AND METHOD: This prospective descriptive and multicenter study (DISCO 3) included 189 patients. Angiographic reperfusion parameters were recorded and resolution of the ST-segment elevation was monitored. Adverse clinical events, such as death, non-fatal reinfarction and repeat revascularization of the culprit vessel, were recorded at discharge, and after one and six months. RESULTS: Direct stenting was performed in 56% of patients, and stenting after predilatation in 44%. The main predictors of direct stenting were short postinfarction delay, non-zero initial TIMI flow, and preinfarction angina. The most common reasons for balloon predilatation were TIMI flow zero on traversing the lesion with a guidewire (92%), involvement of a major bifurcation or tortuous vessel, and severe calcification. Indices of myocardial reperfusion were better with direct stenting: TIMI myocardial perfusion grade 2-3 flow was present in 84% vs 69% (P=.005), and > 70% ST-segment resolution occurred in 66% vs 42% (P=.003). No difference in adverse clinical events was found. CONCLUSIONS: Direct stenting is feasible for treating acute myocardial infarction in more than half of patients. The lesions should not be severely calcified nor involve tortuous vessels, and there should be sufficient flow following passage of a guidewire to define the lesion's characteristics. PMID- 16712746 TI - [Two-year clinical follow-up in 200 patients receiving sirolimus-eluting stents in lesions at a high risk of restenosis]. AB - INTRODUCTION AND OBJECTIVES: Sirolimus-eluting stents (SESs) have been shown to reduce the rate of restenosis significantly in all types of coronary lesion. However, reports of late cases of thrombosis and restenosis have raised questions about long-term outcome in patients treated with these stents. Our aim was to evaluate long-term outcome in patients undergoing SES placement in lesions at a high risk of restenosis. PATIENTS AND METHOD: Since SESs became available, we have used them to treat lesions at risk of restenosis. We studied clinical outcomes in consecutive patients treated with SESs who were followed up for more than 2 years. RESULTS: The study included 200 patients (age 60[11] years, 22% diabetics) who were treated between June 2002 and April 2003 for 309 lesions: 16% were total occlusions, 16.8% in-stent restenoses, 28% diffuse lesions, and 30% small-vessel lesions. The total stent length per patient was 29 (16) mm and the mean diameter was 2.78 (0.27) mm. In a mean clinical follow-up period of 29 (3.2) months (range 24-34 months), there were four deaths, two (1%) of which were cardiac, four (2%) non-fatal infarctions, four (2%) in-stent thromboses (all occurred late, at 3, 7, 26 and 31 months), four (2%) cases requiring target lesion revascularization (at 3, 5, 14 and 15 months), and six (3%) requiring revascularization of a new lesion. CONCLUSIONS: Long-term follow-up of patients undergoing SES placement in lesions at a high risk of restenosis revealed a very low restenosis rate. However, the incidence of late thrombosis appeared to be elevated and warrants further evaluation in larger studies. PMID- 16712747 TI - [Can heart donation exclusion factors be overcome?]. AB - INTRODUCTION AND OBJECTIVES: A shortage of heart donors is limiting the expansion of transplant programs. Our aims were to investigate the impact of different heart donation exclusion factors and to examine ways of increasing the donor pool. PATIENTS AND METHOD: We carried out a retrospective descriptive study of individuals donating organs at a university hospital over a ten-year period. Males under 50 years of age and females under 55 years were regarded as potential heart donors. We recorded the etiology of brain death, initial heart donation exclusion factors, and later reasons for rejection. RESULTS: We studied 130 organ donors, 69 of whom were regarded as potential heart donors. Thirty-nine actually became heart donors (i.e., 30% of all donors and 56.5% of those of a suitable age). Thirteen were excluded because of a history of heart disease; the majority died from ischemic or hemorrhagic stroke, excluding rupture of an aneurysm or arteriovenous malformation (P< .005). Another 11 donors were excluded because of ventricular dysfunction, which was probably secondary to brain death in 10 patients. Ventricular dysfunction accounted for 30% of cases of heart donation exclusion. A comparison of donor subgroups showed that the incidence of ventricular dysfunction did not vary according to the cause of brain death. Among 27 elderly potential donors, 70% died of stroke and 85% had a diagnosis of, or risk factors for, heart disease. CONCLUSIONS: Ventricular dysfunction accounted for 30% of cases of heart donation exclusion. Prevention or reversal of this condition could increase the heart donor pool. PMID- 16712748 TI - [Myocardial disease mortality in children and young adults. A population-based observational study]. AB - INTRODUCTION AND OBJECTIVES: Few studies have investigated death due to myocardial disease in children and young adults. The aim of this study was to analyze the epidemiological, clinical, and pathologic characteristics of death in these cases. PATIENTS AND METHOD: Population-based observational study of all deaths in individuals aged 1-35 years in the Spanish province of Biscay over a period of 12 years. RESULTS: Forty deaths from myocardial disease occurred in 29 males and 11 females (mean age 25.3 years): 30 sudden and 10 non-sudden deaths. The mortality rate was 0.64 per 100,000 persons-year. The relative risk of sudden death was significantly greater than that of non-sudden death, particularly in adolescents and young males. The cause of death was myocarditis in 12 cases (83.3% sudden death), dilated cardiomyopathy in 10 (80% non-sudden death), arrhythmogenic cardiomyopathy in seven, hypertrophic cardiomyopathy in six, and idiopathic concentric left ventricular hypertrophy in five (100% sudden death). Myocardial disease was diagnosed before sudden death in only three cases. Ten subjects had symptoms and electrocardiogram abnormalities but their cardiomyopathy had not been diagnosed. Six individuals had a comorbid condition (morbid obesity in four), six had prodromal symptoms, and 11 had arrhythmic triggering factors (sporting activity in seven). Ventricular fibrillation was frequently observed during cardiopulmonary resuscitation. CONCLUSIONS: Mortality due to myocardial disease in children and young adults is uncommon. Most deaths are sudden. However, some may be preventable. Preventative measures should be aimed at sudden death in adolescents and young males. There was a noticeable association between arrhythmogenic cardiomyopathy and sporting activity. PMID- 16712749 TI - [Inflammation, atherosclerosis and cardiovascular disease risk: PAPP-A, Lp-PLA2 and cystatin C. New insights or redundant information?]. AB - It is well-known that inflammation plays a role in atherogenesis, atherosclerotic plaque progression, and acute coronary syndrome. Inflammatory cells, and cytokines and other biomolecules are implicated in these processes, and have, therefore, been investigated as potential markers of atherosclerotic plaque progression and cardiovascular disease risk. The best characterized and most widely studied is C-reactive protein. However, its role in the clinical setting is still debated. Emerging novel biomarkers that may provide information complementary to that derived from C-reactive protein include pregnancy associated plasma protein A, lipoprotein-associated phospholipase A2, and cystatin C. This article focuses on the potential value of these three new markers in patients with coronary heart disease, and their use as markers of disease risk in apparently healthy individuals. PMID- 16712750 TI - [Cardiovascular disease in women. Why now?]. PMID- 16712751 TI - [Epidemiology of cardiovascular disease in women]. AB - Both mortality due to cerebrovascular disease in individuals aged under 85 years and mortality due to acute myocardial infarction (AMI) are lower in women than men. In contrast, the age-adjusted 28-day case fatality rate after a first AMI is 20% higher in women, particularly in countries where the incidence of AMI is low. In Spain, the case fatality rate is elevated in women hospitalized for a first AMI, but not in those with other forms of coronary heart disease. The pattern of mortality observed after symptom onset, which shows that death is delayed in women, suggests that the mechanism of death is different in the two sexes. The substantial variation that exists in the way results are adjusted and presented make it very difficult to compare the findings of different studies. Relative to men, women with AMI are 10 years older, reach hospital one hour later on average, more frequently have a comorbid condition (mainly diabetes and hypertension), progress to a more serious clinical state, and have a higher adjusted 28-day mortality risk. Moreover, the treatment given to women during the acute phase is less aggressive. A change in healthcare workers' attitudes is needed so that women with acute coronary syndromes can be identified earlier, thereby increasing the use of diagnostic and therapeutic procedures to a level that corresponds to the greater severity of AMI observed in women at presentation. PMID- 16712752 TI - [Virtual histology of a post-angioplasty intramural hematoma]. PMID- 16712753 TI - [Mortality and morbidity in HIV-infected patients undergoing coronary artery bypass surgery: a case control study]. AB - The use of highly active antiretroviral therapy (HAART) in patients with HIV infection has improved survival. This improvement combined with the metabolic effects of treatment has increased cardiovascular risk and the need for cardiac surgery in these patients. We compared morbidity and mortality in HIV-infected patients (cases, n=7) and non-HIV-infected patients (controls, n=21) who underwent isolated coronary artery surgery between 1997 and 2004. The durations of extracorporeal circulation and aortic cross-clamping were shorter in HIV infected patients (P=.002 and P=.014, respectively). The percentage of patients who experienced complications was similar, at 57.1% in both groups, but there was a slightly higher number of complications per patient in non-HIV-infected individuals. The mean length of total hospitalization was greater in HIV-infected patients (27.1 [13.3] versus 8.8 [5.3] days; P=.003), as was that of postoperative hospitalization (18.2 [15.4] vs 7.9 [4.2] days; P=.08). No HIV infected patient died or needed a repeat cardiac operation. No progression of the HIV infection was observed. Isolated coronary artery surgery in HIV-infected patients produces good results, and there is no increase in morbidity or mortality. Extracorporeal circulation did not influence disease progression. PMID- 16712754 TI - [Prognostic role of new biomarkers in off-pump coronary artery bypass surgery]. AB - The value of biomarkers of myocardial damage or inflammation in off-pump coronary artery bypass (OPCAB) surgery has not yet been established. In a prospective study of 51 consecutive patients scheduled for elective OPCAB surgery, preoperative levels of troponin T, C-reactive protein, interleukin-6 and tumor necrosis factor-alpha were determined. The primary endpoint was the combination of cardiac death or acute myocardial infarction (AMI) within 30 days. Seven patients (14%) presented with an adverse event: three cardiac deaths and six AMIs. Univariate analysis identified the following adverse event predictors: renal failure (50% vs 11%, P=.028), left ventricular ejection fraction 0.10 ng/dL (43% vs 9%, P=.016), and EuroSCORE rating (7.6 [2.5] VS. 5.2 [2.6], P=.031). A preoperative troponin-T level > 0.10 ng/dL (P=.03) was the only independent adverse event predictor. No significant differences were found with biomarkers of inflammation (P.05). The presence of a preoperative troponin-T level > 0.10 ng/dL is associated with a higher risk of cardiac death or AMI in patients undergoing OPCAB surgery. PMID- 16712755 TI - [Clinical and echocardiographic characteristics of scimitar syndrome]. AB - Seven patients with scimitar syndrome underwent retrospective clinical and echocardiographic examination. The findings were: respiratory infection with dyspnea on moderate exercise in 90%, scimitar sign in four (57%), dextrocardia in five (71%), and interatrial septal defect in five (71%), one of whom had patent ductus arteriosus. Overall, two patients had patent ductus arteriosus: one also had aortic coarctation and the other, a bicuspid aortic valve. Dilation of the right cavities was found in five (71%) and blunt edge in five (71%). In two patients, anomalous drainage was into the right atrium; in another two, into the inferior vena cava; and in three, towards the junction of the right atrium and the inferior vena cava. In three patients, drainage was obstructed. Six patients with cardiac abnormalities proceeded to surgery. Scimitar syndrome is a rare entity. In the large majority of patients (86%), diagnosis and follow-up can be performed echocardiographically. PMID- 16712756 TI - [Pericardial constriction secondary to cholesterol pericarditis]. PMID- 16712757 TI - [Managing heart failure in the elderly: a primary-care perspective]. PMID- 16712758 TI - Influence of visibility out of the vehicle cabin on lane-change crashes. AB - The influence of lateral visibility from the vehicle cabin on safety was evaluated by examining the differences in lane-change crashes of four-door and two-door body styles of the same vehicle models. These two vehicle styles were used because B-pillars (the pillars between the front and rear seats) on four door models are farther forward, and thus nearer the fore-aft position of the driver. (Furthermore, the B-pillars on two-door models can be narrower, and some two-door models have no B-pillars at all.) To control for driver differences between these two body styles, going-straight-ahead crashes were used for comparison. The analysis used 2000-2003 North Carolina crash data, and considered the crash experience of four-door and two-door body styles for the same 10 vehicles for model years 1995 and newer. The main finding is that four-door body styles are more likely to be involved in lane-change crashes than are two-door body styles of the same vehicle models. This finding suggests that lateral visibility out of the vehicle cabin affects safety. PMID- 16712759 TI - Association between setting quantified road safety targets and road fatality reduction. AB - Since the 1980s, many countries have set quantified road safety targets as part of their comprehensive road safety strategies. This paper studies the association between the setting of quantified road safety targets and road fatality reduction over the past two decades. Both aggregate and disaggregate levels of hypothesis tests are performed on data from 14 countries that set road safety targets during the period 1981-1999. A before-and-after analysis of the treatment and comparison groups is conducted, and the results show that the majority of countries with quantified road safety targets experienced a reduction in road fatalities in this period. Overall, the establishment of quantified road safety targets is found to have an appreciable association with an improvement in road safety. PMID- 16712760 TI - Estimation of reduced life expectancy from serious occupational injuries in Taiwan. AB - This study assesses the loss of life expectancy and potential working life in Taiwan, between 1986 and 2000, from various types of occupational accidents resulting in mortality or permanent disability. The databases on occupational mortality and permanent disability were obtained from the Bureau of Labor Insurance, with this information then being linked to national death registry data to construct the survival function. A Monte Carlo simulation method was subsequently used to extrapolate the survival rate for up to 600 months, to derive the life expectancy for different types of occupational accidents leading to permanent disability (n=81,249). Based upon the life table for each calendar year, the life expectancy lost by age-gender cohort was also estimated for cases of mortality (n=20,001). In those cases resulting in permanent occupational disability, variations in the expected years of life lost (EYLL) were demonstrated by different occupational injuries, ranging from 7.4 to 13.6 years per case. The overall EYLL through permanent occupational disability is found to be almost identical to that of occupational mortality, with a ratio of 1.04:1. We conclude that permanent disability resulting from occupational injuries has a significant impact on society as a whole. PMID- 16712761 TI - Cross-cultural differences in driving skills: a comparison of six countries. AB - The first aim of the present study was to investigate the applicability of the two-factor structure (perceptual-motor skills by 11 items, e.g., "fluent driving"; safety skills by 9 items, e.g., "conforming to the speed limits") of the Driver Skill Inventory (DSI) among British, Dutch, Finnish, Greek, Iranian, and Turkish drivers. It was also hypothesized that the combination of self reported high ratings of perceptual-motor skills and low ratings of safety skills creates a serious risk for dangerous driving and road accident involvement. The second aim of this study was, therefore, to investigate this asymmetric relationship between perceptual-motor and safety skills in traffic penalties and accident involvement. Two hundred and forty two drivers were chosen from each of the six countries, matched for age and sex. The results of exploratory factor analyses together with target rotation showed that the two-factor structure of DSI found in "safe" Northern and Western European countries were highly congruent. However, the safety skills factor of DSI in Greece, Iran, and Turkey was relatively incongruent in spite of high factor similarity found in perceptual motor skills. The asymmetric relationship between perceptual-motor and safety skills on traffic penalties was found in Finland and Turkey. A negative relationship between safety skills and the number of accidents was found both in Greece and Iran while a positive relationship between perceptual-motor skills and the number of accidents was found only in Iran. PMID- 16712762 TI - Nanomechanical measurements with AFM in the elastic limit. AB - With increasing interest in nanoscience and nanotechnology, the fundamental underpinnings of what makes materials strong and durable are under critical investigation. Recent findings suggest that when materials are reduced in extent to nanoscopic proportions, they exhibit enhanced strength, specifically in the form of higher moduli than are measured on macroscopic objects of the same composition. Force-deformation behavior of nanostructures subjected to concentrated loads, such as with atomic force microscopy (AFM), can yield detailed information and insight about their local mechanical properties. We review and evaluate the effectiveness of deformation and indentation tests used in determining the elastic modulus of nanobeams, nanosprings, thin films, biological samples, dendrimers, and fluid droplets. Obstacles yet remain in the determination of absolute, quantitative modulus data at the nanoscale. In spite of basic limitations, recent developments in advanced nanomechanical techniques will facilitate improvement in our understanding of material strength and aging from molecules and colloids to the macroscale. PMID- 16712764 TI - Monitoring viral decontamination procedures with green fluorescent protein expressing adenovirus. PMID- 16712763 TI - Payne rearrangement during analysis of epoxyalcohols of linoleic and alpha linolenic acids by normal phase liquid chromatography with tandem mass spectrometry. AB - Hydroperoxides of polyunsaturated fatty acids can be transformed to epoxyalcohols and keto fatty acids by metal enzymes, hematin, and various catalysts. In the current study, we used hematin to transform 9-hydroperoxy-10E,12Z-octadecadienoic acid and 13-hydroperoxy-9Z,11E-octadecadienoic acid to epoxyalcohols (with trans epoxide configuration) and to keto fatty acids. The products were separated by normal phase high-performance liquid chromatography (NP-HPLC) and analyzed using postcolumn addition of isopropanol/water and online negative ion electrospray ionization mass spectrometry (MS). The tandem MS (MS/MS) spectra were studied using analogs prepared from [9,10,12,13-2H4]linoleic acid (18:2n-6) and from alpha-linolenic acid (18:3n-3). We also studied the MS/MS spectra of epoxyalcohols formed from 11-hydroperoxy- and 8-hydroperoxy-9Z,12Z octadecadienoic acids. Results were confirmed by MS/MS analysis of a series of authentic standards. MS/MS ions of 9-keto-10E,12Z-octadecadienoic acid and 13 keto-9Z,11E-octadecadienoic acid could be explained by keto-enol tautomerism. MS/MS spectra of regioisomeric allylic epoxyalcohols differed in relative intensities of characteristic ions. The MS/MS spectra of the epoxyalcohols with 1 hydroxy-2,3-epoxy-4Z-pentene or 3-hydroxy-1,2-epoxy-4Z-pentene elements were virtually identical and showed two characteristic ions that differed by 30 in m/z values (CH(OH)). The results suggested that epoxide migration (Payne rearrangement) occurred during collision-induced dissociation. We conclude that regioisomeric allylic epoxyalcohols can be identified by their MS/MS spectra, whereas regioisomeric epoxyalcohols can be identified by MS/MS in combination with their retention times on NP-HPLC. PMID- 16712765 TI - An activity-based two-hybrid system for the selective identification of substrates of protein kinases. PMID- 16712766 TI - Capillary electrophoresis-mass spectrometry as a characterization tool for therapeutic proteins. AB - With the increasing use of capillary electrophoresis (CE) in the biotechnology industry, there is a demand for analytical tools and methodology that can be used to characterize CE profiles. This article describes the implementation and optimization of a robust online CE-mass spectrometry (CE-MS) system used for the characterization of several CE assays developed at Genentech Inc. These assays include CE as a complement to reverse-phase peptide mapping for the identification of small peptides eluting in the void volume, profiling N-linked glycopeptide heterogeneity, and determining O-linked site occupancy. In addition, CE-MS was used to confirm major 8-aminopyrene-1,3,6-trisulfonate (APTS)-labeled glycans released from recombinant antibodies that are routinely profiled by CE laser-induced fluorescence (CE-LIF). For each study, CE-MS was able to successfully identify components seen in UV or LIF electropherograms, thereby expanding the capability of CE and CE-MS for profiling biomolecules. PMID- 16712767 TI - Dual fluorescent halogen immunoassay for bioaerosols using confocal microscopy. PMID- 16712769 TI - [Mesenteric hematoma: Unusual complication of a long term oral anticoagulation therapy]. AB - Authors report a case of a 77 years old man who developed a small bowel mesenteric hematoma as consequence of an unusual complication from a long term oral anticoagulation treatment. Computed tomoangiography helpfully suggests diagnosis of a mesenteric hematoma and refuts an organic cause as vascular anomalies, by an equivalent method to conventional angiography, noninvasely. In our case report, patient's deterioration justified an emergency surgery corroborating medical imager findings. Authors review the rare cases previously reported. PMID- 16712768 TI - Flow injection analysis-Rayleigh light scattering detection for online determination of protein in human serum sample. AB - A flow injection analysis (FIA) system combined with Rayleigh light scattering (RLS) detection is developed for the sensitive and rapid determination of protein concentration in human serum sample. This method is based on the weak intensity of RLS of Eriochrome Black T (EBT, 2-hydroxy-1-(1-hydroxy-2-naphthylazo)-6 nitronaphthalene-4-sulfonic acid sodium salt), which can be enhanced by the addition of protein in weakly acidic solution. The effects of pH and interfering species on the determination of protein were examined. Calibrations for protein, based on RLS intensity, were linear in the concentration ranges of 7-36 microg/ml for human serum album (HSA) and 8-44 microg/ml for bovine serum album (BSA). The detection limits of the method were found to be 0.882 and 2.507 microg/ml for HSA and BSA, respectively. A relative standard deviation of 0.76% (n=5) was obtained with 20 microg/ml HSA standard solution. The FIA-RLS method was more stable than the general RLS method, and the average RSD value of FIA-RLS was less than that of the general RLS. The sample rate was determined to be 90 samples per hour. PMID- 16712770 TI - [Anal adenocarcinoma revealed by a fistula-in-ano. Report of a case]. AB - Anal adenocarcinoma revealed by a fistula-in-ano occurs rarely. Authors report a case in a 56 year-old man. The pathogeny is still unknown. Symptomatology has no specificity and the diagnosis is often late, in an advanced stage of the sickness. Recurrent or non recurrent fistula-in-ano requires multiple biopsies for pathology analysis in order to screen a related cancer. Tumor dimension, lymph node involvement, histologic grade and treatment modality are independent prognosis factors for survival. Our objective is to show physiopathological, clinical and therapeutical characteristics from a review of the literature. PMID- 16712771 TI - Energetics of membrane protein folding and stability. AB - The critical role of membrane proteins in a myriad of biological and physiological functions has spawned numerous investigations over the past several decades with the long-term goal of identifying the molecular origins and energetic forces that stabilize these proteins within the membrane. Parallel structural and thermodynamics studies on several systems have provided significant insight regarding the driving forces governing folding, assembly, insertion, and translocation of membrane proteins. The present review surveys families of membrane-associated proteins including alpha-helical and beta-barrel structures, viral surface receptors, and pore-forming toxins, citing representative proteins within each of these classes for further scrutiny in terms of structure-function relationships and global conformational stability. This overview presents seminal findings from pioneering studies on the energetics of membrane protein folding and stability to modern techniques that are exploiting the use of molecular genetics and single molecule studies. An overall consensus regarding the molecular origins of membrane protein stability is that a number of intrinsic properties resemble features of soluble proteins, yet there are distinct energetic differences arising from specific intra- and intermolecular interactions within the membrane. The combined efforts from structural, energetics, and dynamics approaches offer unique insights and improve our fundamental understanding of the driving forces dictating membrane protein folding and stability. PMID- 16712772 TI - Thr373, Asp375, and Lys260 are in the coenzyme site of porcine NADP-dependent isocitrate dehydrogenase. AB - Thr(373), Lys(374), Asp(375), and Lys(260) were chosen as site-directed mutagenesis targets within porcine NADP-dependent isocitrate dehydrogenase based on structurally corrected sequence alignment among prokaryotic and eukaryotic NADP-isocitrate dehydrogenases. Wild-type and all mutant enzymes were expressed in Escherichia coli and purified to homogeneity. These mutations do not alter the secondary structure or dimerization state of the mutants. The D375N and K260Q mutants exhibit, respectively, a 15- and 28-fold increase in K(m) for NADP, along with marked decreases in V(max) as compared to wild-type enzyme. In contrast, replacing Lys(374), which was previously proposed to contribute to apparent coenzyme affinity, does not change the enzyme's kinetic parameters. T373S exhibits similar kinetic parameters to those of wild-type while T373A and T373V mutations reduce the V(max) values of the resulting enzymes to 1 and 20%, respectively of that of wild-type. We conclude that a hydroxyl group at position 373 is required for effective enzyme function and that Asp(375) and Lys(260) are critical amino acids contributing to coenzyme affinity as well as catalysis by porcine NADP-isocitrate dehydrogenase. PMID- 16712773 TI - Identification of critical arginine residues in the functioning of Rubisco activase. AB - Rubisco activase is a member of the AAA(+) family in which arginines located in the Box VII and Sensor 2 domains are a recurrent feature and typically contribute to ATP-binding/hydrolysis or an inter-subunit interface. Replacement of R241 or R244 in Box VII or R294 or R296 in Sensor 2 with alanine in tobacco activase did not greatly alter the binding of ATP or ADP. However, ATP hydrolysis was minimal (R241A and R244A) or greatly diminished (R296A) and none of these mutants were able to activate Rubisco. R241, R244 and R296 were also required for nucleotide dependent conformational changes detected by intrinsic fluorescence and limited proteolysis. ATP-induced oligomerization, monitored by gel filtration, was not observed with the wild type and mutant tobacco activases in contrast to spinach activase and a R239A mutant (corresponding to R244A in tobacco). Thus, there is not a strict correlation of oligomerization with ATP hydrolysis and intrinsic fluorescence. PMID- 16712774 TI - Ca-binding to Bacillus licheniformis alpha-amylase (BLA). AB - Ca-induced renaturation of Bacillus licheniformis alpha-amylase in the presence of urea has been employed to determine the binding constants of the ion. The native enzyme is folded at 3M urea while the Ca-depleted protein is largely unfolded at this denaturant concentration. Refolding of the protein has been monitored by circular dichroism and the titration curves have been analyzed assuming a model of three independent binding sites. The stoichiometry has been taken from X-ray studies. The refolded protein exhibits a secondary structure that is similar but not identical to that of the native protein. The binding constants have been used to construct a phase diagram that illustrates the contribution of Ca-binding to the resistance against urea unfolding. PMID- 16712775 TI - Magnesium and the inflammatory response: potential physiopathological implications. AB - The purpose of this review is to summarize experimental findings showing that magnesium modulates cellular events involved in inflammation. Experimental magnesium deficiency in the rat induces after a few days a clinical inflammatory syndrome characterized by leukocyte and macrophage activation, release of inflammatory cytokines and acute phase proteins, excessive production of free radicals. Increase in extracellular magnesium concentration, decreases inflammatory response while reduction in the extracellular magnesium results in cell activation. Because magnesium acts as a natural calcium antagonist, the molecular basis for inflammatory response is probably the result of modulation of intracellular calcium concentration. The priming of phagocytic cells, the opening calcium channel and activation of N-methyl-d-aspartate (NMDA) receptors, the activation of nuclear factor-kappa B (NFkappaB) have been considered as potential mechanisms. Moreover, magnesium deficiency induces a systemic stress response by activation of neuro endocrinological pathways. As nervous and immune systems interact bidirectionally, the roles of neuromediators have also been considered. Magnesium deficiency contributes to an exaggerated response to immune stress and oxidative stress is the consequence of the inflammatory response. Inflammation contributes to the pro-atherogenic changes in lipoprotein metabolism, endothelial dysfunction, thrombosis, hypertension and explains the aggravating effect of magnesium deficiency on the development of metabolic syndrome. Further studies are still needed to assess more accurately the role of magnesium in immune response in humans, but these experimental findings in animal models suggest that inflammation is the missing link to explain the role of magnesium in many pathological conditions. PMID- 16712776 TI - A study of the topoisomerase II activity in HIV-1 replication using the ferrocene derivatives as probes. AB - Human Topoisomerase II is present in two isoforms, 170KDa alpha and 180KDa beta. Both the isoforms play a crucial role in maintenance of topological changes during DNA replication and recombination. It has been shown that Topoisomerase II activity is required for HIV-1 replication and the enzyme is phosphorylated during early time points of HIV-1 replication. In the present study, we have studied the molecular action of Topoisomerase II inhibitors, azalactone ferrocene (AzaFecp), Thiomorpholide amido methyl ferrocene (ThioFecp), and Ruthenium benzene amino pyridine (Ru(ben)Apy) on cell proliferation and also on various events of HIV-1 replication cycle. The Topoisomerase II beta over-expressing neuroblastoma cell line shows a higher sensitivity to these compounds compared to the Sup-T1 cell line. All the three Topoisomerase II inhibitors show significant anti-HIV activity at nanomolar concentrations against an Indian isolate of HIV 1(93IN101) in Sup-T1 cell line. An analysis of action of these compounds on proviral DNA synthesis at 5h of post-infection shows that they inhibit proviral DNA synthesis as well as the formation of pre-integration complexes completely. Further analysis, using polymerase chain reaction and western blot, showed that both the Topoisomerase II alpha and beta isoforms are present in the pre integration complexes, suggesting their significant role in HIV-1 replication. PMID- 16712777 TI - A comparative study of systemic subantimicrobial and topical treatment of minocycline in experimental periodontitis of rats. AB - OBJECTIVE: The purpose of this study was to compare the effectiveness of minocycline on treating experimentally induced periodontitis in rats when administered either as a systemic subantimicrobial dose or as a topical ointment. DESIGN: Thirty-two adult male Sprague-Dawley rats in four experimental groups-(1) model group; (2) systemic subantimicrobial dose of minocycline (5mg/kg/day) treatment group; (3) topical subgingival dose of minocycline (2mg/animal/week) treatment group; (4) control group. Experimentally induced periodontitis-silk ligatures were placed around the crevices of the second molar teeth and the animals fed a 10% sucrose drink. Assessment was carried out at days 28 and 56 using a number of different visual, histological and ultrastructure approaches. (1) Visual assessment-tooth mobility, gingival index and alveolar bone loss. (2) Histological examination-monocyte infiltration and resorption lacunae with osteoclasts. (3) Transmission electron microscopy (TEM)-morphological transformation of fibroblasts and osteoclasts. The collected data were analysed for statistical significance using the analysis of variance statistical test. RESULTS: Minocycline significantly reduced tooth mobility, gingival index and alveolar bone loss when administered either systemically or as a topical ointment compared to the model group (P<0.01). However, the alveolar bone loss was significantly less (P<0.01 in the systemic treatment group compared to the local treatment group. Monocyte infiltration and resorption lacunae with osteoclasts were significantly less in the both treatment groups compared to the model group (P<0.01). The osteoclasts failed to form a ruffled border in the systemic treatment group. CONCLUSION: Topical treatment significantly reduces gingivitis while systemic treatment is beneficial in terms of inhibiting alveolar bone loss. PMID- 16712778 TI - Mitochondrial subpopulations and heterogeneity revealed by confocal imaging: possible physiological role? AB - Heterogeneity of mitochondria has been reported for a number of various cell types. Distinct mitochondrial subpopulations may be present in the cell and may be differently involved in physiological and pathological processes. However, the origin and physiological roles of mitochondrial heterogeneity are still unknown. In mice skeletal muscle, a much higher oxidized state of subsarcolemmal mitochondria as compared with intermyofibrillar mitochondria has been demonstrated. Using confocal imaging technique, we present similar phenomenon for rat soleus and gastrocnemius muscles, where higher oxidative state of mitochondrial flavoproteins correlates also with elevated mitochondrial calcium. Moreover, subsarcolemmal mitochondria demonstrate distinct arrangement and organization. In HL-1 cardiomyocytes, long thread mitochondria and small grain mitochondria are observed irrespective of a particular cellular region, showing also heterogeneous membrane potential and ROS production. Possible physiological roles of intracellular mitochondrial heterogeneity and specializations are discussed. PMID- 16712779 TI - Rescue of F508del CFTR: Commentary on "F508del CFTR with two altered RXR motifs escapes from ER quality control but its channel activity is thermally sensitive". PMID- 16712780 TI - Aquaporin-5 water channel in lipid rafts of rat parotid glands. AB - Aquaporin-5 (AQP5), an apical plasma membrane (APM) water channel in salivary glands, lacrimal glands, and airway epithelium, has an important role in fluid secretion. The activation of M3 muscarinic acetylcholine receptors (mAChRs) or alpha1-adrenoceptors on the salivary glands induces salivary fluid secretion. AQP5 localizes in lipid rafts and activation of the M3 mAChRs or alpha1 adrenoceptors induced its translocation together with the lipid rafts to the APM in the interlobular ducts of rat parotid glands. This review focuses on the mechanisms of AQP5 translocation together with lipid rafts to the APM in the interlobular duct cells of parotid glands of normal rats and the impairment of AQP5 translocation in diabetes and senescence. PMID- 16712781 TI - Memory of childhood trauma before and after long-term psychological treatment of borderline personality disorder. AB - The present study investigated the consistency of self-reports of childhood traumatic events in a sample of 50 patients with a borderline personality disorder (BPD) before and after 27 months of intensive treatment with schema focused therapy or transference focused psychotherapy. The mean number of reported sexual, physical and emotional traumatic events did not change following treatment. Test-retest correlations of the trauma-interview also indicated high stability of the total number of sexual, physical and emotional events reported. The majority of the patients, however, did no longer report at least one of the 33 listed events after psychotherapy, and the majority reported at least one event that they had not mentioned before the start of treatment. These findings were not related to type of treatment or changes in suppression, intrusions, avoidance of intrusions, dissociative symptoms, depressive symptoms, and borderline symptoms. PMID- 16712782 TI - Effects of temptations on the affective salience of weight control goals. AB - Despite the value of weight control goals, the maintenance of healthy eating habits represents a challenge for most. Self-regulatory efforts are often challenged by the presence of high-risk cues (e.g., tempting foods) which provide short-term positive outcomes at the expense of these long-term health objectives. The current study examined contextual influences on self-regulation failure by exploring the effect of cues on an indirect measure of goal value. Two experiments were conducted with undergraduate students which examined the effect of temptation cues on the evaluation of information related to the goal of weight control. Results of Study 1 provided preliminary evidence for the utility of this task as an indirect measure of goal value and showed that food-related primes slowed evaluation response times for weight control-related targets. Study 2 replicated and extended these findings by demonstrating that temptation cues may not only decrease the affective salience of weight control related information but increase the salience of information related to the goal of affect enhancement. These results suggest that self-regulation failure may be influenced by contextual changes in the value of health-related goals. Implications for prevention and intervention efforts are discussed. PMID- 16712783 TI - PTSD symptom trajectories: from early to chronic response. AB - This study aimed to identify posttraumatic stress disorder (PTSD) symptom trajectories across the first 12 months following traumatic injury. Three hundred and seven consecutively admitted injury survivors were assessed for severity of PTSD symptoms just prior to discharge, and at 3 and 12 months postinjury. Growth modeling was used to determine the curve that best fit the trajectory for each symptom cluster over the 12-month period. Individuals with 12-month PTSD showed significantly higher re-experiencing, arousal, and avoidance symptoms at eight days posttrauma relative to those without, and these symptoms escalated over time. Those without PTSD maintained their relatively low symptom levels. These findings highlight that individuals who will go onto develop PTSD have a distinctly different symptom course than those who recover. PMID- 16712784 TI - Smoking motives in the prediction of affective vulnerability among young adult daily smokers. AB - The primary aim of this study was to examine whether smoking to reduce negative affect was uniquely related to a range of affective vulnerability factors (e.g., anxiety sensitivity, anxious arousal, and negative affectivity) among daily smokers. Participants were 276 young adult daily smokers (124 females; M(age)=25.12, SD=10.37). Partially consistent with prediction, the motivation to smoke to reduce negative affect was significantly related to anxiety sensitivity and negative affectivity, but not anxious arousal; the observed significant effects were above and beyond other theoretically relevant factors (e.g., smoking rate, years smoked, age, gender). In contrast to prediction, habitual smoking motives demonstrated significant incremental associations with anxiety sensitivity and anxious arousal symptoms. These results suggest that there are important associations between certain smoking motives and negative affective states and that such relations are not attributable to other smoking factors (e.g., smoking rate). PMID- 16712785 TI - Chlamydial DNA polymerase I can bypass lesions in vitro. AB - We found that DNA polymerase I from Chlamydiophila pneumoniae AR39 (CpDNApolI) presents DNA-dependent DNA polymerase activity, but has no detectable 3' exonuclease activity. CpDNApolI-dependent DNA synthesis was performed using DNA templates carrying different lesions. DNAs containing 2'-deoxyuridine (dU), 2' deoxyinosine (dI) or 2'-deoxy-8-oxo-guanosine (8-oxo-dG) served as templates as effectively as unmodified DNAs for CpDNApolI. Furthermore, the CpDNApolI could bypass natural apurinic/apyrimidinic sites (AP sites), deoxyribose (dR), and synthetic AP site tetrahydrofuran (THF). CpDNApolI could incorporate any dNMPs opposite both of dR and THF with the preference to dAMP-residue. CpDNApolI preferentially extended primer with 3'-dAMP opposite dR during DNA synthesis, however all four primers with various 3'-end nucleosides (dA, dT, dC, and dG) opposite THF could be extended by CpDNApolI. Efficiently bypassing of AP sites by CpDNApolI was hypothetically attributed to lack of 3' exonuclease activity. PMID- 16712786 TI - Gliotoxin induces caspase-dependent neurite degeneration and calpain-mediated general cytotoxicity in differentiated human neuroblastoma SH-SY5Y cells. AB - In this study, a significant increase by 50% in intracellular free calcium concentration ([Ca(2+)](i)) was observed in differentiated human neuroblastoma (SH-SY5Y) cells after exposure to 0.25microM of the fungal metabolite gliotoxin for 72h. Further, the involvement of caspases and calpains was demonstrated to underlie the gliotoxin-induced cytotoxic and neurite degenerative effects. The caspase inhibitor Z-VAD-fmk almost completely reduced the neurite degeneration from 40% degeneration of neurites to 5% as compared to control. Inhibition of calpains with calpeptin significantly attenuated gliotoxin-induced cytotoxicity, determined as reduction in total cellular protein content, from 43% to 14% as compared to control cells. Western blot analyses of alphaII-spectrin breakdown fragments confirmed activity of the proteases, and that alphaII-spectrin was cleaved by caspases in gliotoxin-exposed cells. These results show that calpains and caspases have a role in the toxicity of gliotoxin in differentiated SH-SY5Y cells and that the process may be Ca(2+)-mediated. PMID- 16712787 TI - STMN2 is a novel target of beta-catenin/TCF-mediated transcription in human hepatoma cells. AB - The activity of beta-catenin/TCF, the key component of Wnt signaling pathway, is frequently deregulated in human cancers, resulting in the activation of genes whose dysregulation has significant consequences on tumor development. Therefore, identifying the target genes of Wnt signaling is important for understanding beta catenin-mediated carcinogenesis. Here, we report STMN2, a gene implicated in the regulation of microtubule dynamics, as a novel target of beta-catenin-mediated transcription. STMN2 was up-regulated in hepatoma and cirrhotic liver tissues compared to normal liver and also in cell lines where beta-catenin/TCF is constitutively activated. Transient activation of beta-catenin/TCF either by transfection of a constitutively active form of beta-catenin or by LiCl treatment induced the STMN2 mRNA expression in PLC/PRF/5 cells. Of the four members of STMN gene family, only STMN2 showed a Wnt-dependent expression pattern. Through promoter mapping and chromatin immunoprecipitation assays, we found that STMN2 is a direct target of beta-catenin/TCF-mediated transcription and that the TCF binding site at -1713 of STMN2 promoter is critical for beta-catenin/TCF dependent expression regulation. siRNA-mediated knock-down of STMN2 expression indicated that STMN2 is required for maintaining the anchorage-independent growth state of beta-catenin/TCF-activated hepatoma cells. Our results suggest that STMN2 might be a novel player of beta-catenin/TCF-mediated carcinogenesis in the liver. PMID- 16712788 TI - Augmentation of monocyte intracellular ascorbate in vitro protects cells from oxidative damage and inflammatory responses. AB - Ascorbic acid is present as a primary antioxidant in plasma and within cells, protecting both cytosolic and membrane components of cells from oxidative damage. The effects of intracellular ascorbic acid on F(2)-isoprostanes (biomarkers of oxidative stress) and monocyte chemoattractant protein-1 (marker of inflammatory responses) production in monocytic THP-1 cells were investigated under conditions of 2,2'-Azobis(2-methylpropionamidine)dihydrochloride (AAPH) induced oxidative stress. Cells cultured under normal conditions have extremely low ascorbate levels and the intracellular ascorbate can be augmented significantly by adding ascorbate to the culture medium. While AAPH treatment reduced cell viability, increased F(2)-isoprostanes and MCP-1 production, the presence of intracellular ascorbic acid maintained high cell viability and attenuated both F(2) isoprostanes and MCP-1 production. Measurement of intracellular ascorbic acid and its oxidised products showed that intracellular ASC was oxidised to a significantly greater extent during AAPH treatment and may be utilised to protect the cells under conditions of oxidative stress. This study demonstrates the importance of intracellular ascorbate, which may be lacking under normal cell culture conditions, under conditions of increased oxidative stress. PMID- 16712789 TI - Association of Polycomb group SUZ12 with WD-repeat protein MEP50 that binds to histone H2A selectively in vitro. AB - SUZ12 is a Polycomb group protein that forms Polycomb repressive complexes (PRC2/3) together with EED and histone methyltransferase EZH2. Although the essential role of SUZ12 in regulating the activity of the PRC2/3 complexes has been demonstrated, additional function of this protein was suggested. Here, we show that SUZ12 interacts with WD-repeat protein MEP50 in vitro and in vivo. We show that the MEP50 binds histone H2A selectively among core histones, and mediates transcriptional repression of protein arginine methyltransferase PRMT5, which is known to methylate H2A and H4. These results suggest that SUZ12 might have a role in transcriptional regulation through physical interaction with MEP50 that can be an adaptor between PRMT5 and its substrate H2A. PMID- 16712790 TI - Alpha-bulnesene, a novel PAF receptor antagonist isolated from Pogostemon cablin. AB - Alpha-bulnesene is a sesquiterpenoid isolated from the water extract of Pogostemon cablin. It showed a potent and concentration-dependent inhibitory effect on platelet-activating factor (PAF) and arachidonic acid (AA) induced rabbit platelet aggregation. In a radioligand binding assay for the PAF receptor, alpha-bulnesene competitively inhibited [(3)H]PAF binding to the PAF receptor with an IC(50) value of 17.62+/-5.68microM. alpha-Bulnesene also dose-dependently inhibited PAF-induced intracellular Ca(2+) increase in fluo-3/AM-loaded platelets (IC(50) values of 19.62+/-1.32microM). Furthermore, alpha-bulnesene inhibited AA induced thromboxane B(2) (TXB(2)) formation and prostaglandin E(2) (PGE(2)) formation. These results indicate that the inhibitory effect of alpha-bulnesene on platelet aggregation was due to a dual activity; specifically the chemical blocked PAF-induced intracellular signal transduction and interfered with cyclooxygenase activity, which resulted in a decrease in thromboxane formation. This study is the first to demonstrate that alpha-bulnesene is a PAF receptor antagonist as well as an anti-platelet aggregation agent. PMID- 16712792 TI - Identification of pseudo attP sites for phage phiC31 integrase in bovine genome. AB - Streptomyces phage phiC31 integrase was found to mediate site-specific integration of foreign genes at pseudo attP sites of genomes in human, mouse, rat, and Drosophila. This paper reports that phiC31 integrase can also mediate homologous recombination between attB and pseudo attP sites in bovine cells and foreign gene integration was increased at least 2-fold in bovine fibroblasts or Madin-Darby bovine kidney (MDBK) cells. Two intrinsic pseudo attP sites named BpsF1 and BpsM1 located in the inter-gene regions on chromosome 28 and 19, respectively, were identified in bovine genome. These pseudo attP sites shared similar characteristics with those from other species as previously described. Our study demonstrated that the phiC31 integrase system provides a new potential for genetic engineering of the bovine genome and might be beneficial for the research on ruminant. PMID- 16712793 TI - Pim-1 kinase expression during murine mammary development. AB - Pim-1 kinase phosphorylates substrates whose activities are linked to proliferation, survival, differentiation, and apoptosis. Although pim-1 is induced by hormones and cytokines, the hormonal control and contribution of Pim-1 to mammary gland development have not been evaluated. We examined Pim-1 expression in mammary cell lines, investigated whether Pim-1 levels could be altered in breast epithelia by mammogenic hormones, and evaluated Pim-1 expression during mammary development. We found that Pim-1 was elevated in most mammary carcinoma cell lines and progesterone increased Pim-1 protein to some extent in non-tumorigenic mammary epithelia. Pim-1 expression in situ was consistent with the documented profile of progesterone activity in mouse mammary glands. Pim-1 nuclear localization correlated with cytoplasmic distribution for its substrate, p21(CIP/Waf1), and we found that Pim-1 and p21 associate in vitro. Our results suggest that Pim-1 expression may be regulated by progesterone during mammary development and Pim-1 associates with p21 in mammary epithelial cells. PMID- 16712794 TI - Co-culture with fat cells induces cellular insulin resistance in primary hepatocytes. AB - Obesity is highly correlated with systemic insulin resistance. To assess the effect of fat cell on the development of hepatic insulin resistance, an in vitro system was developed in which primary hepatocytes were kept in co-culture with 3T3-L1 cells, then insulin signaling and glycogen production were subsequently analyzed in hepatocytes. The results showed that insulin-induced tyrosine phosphorylation of insulin receptor substrate (IRS)-2 was significantly blocked. Insulin-regulated activation of Akt kinase and glucose production in the hepatocytes were also reduced after co-culture. On the other hand, addition of TNF-alpha or IL-6 neutralizing antibodies to the supernatant of co-culture recovered both IRS-2 phosphorylation and Akt activation. In conclusion, fat cells may induce insulin resistance in liver cells, and this process appears to be mediated by TNF-alpha and IL-6. Our data present first the direct evidence of interaction for insulin signaling event between the adipocytes and hepatocytes. PMID- 16712795 TI - Beyond gastric acid reduction: proton pump inhibitors induce heme oxygenase-1 in gastric and endothelial cells. AB - Proton pump inhibitors (PPIs) have been demonstrated to prevent gastric mucosal injury by mechanisms independent of acid inhibition. Here we demonstrate that both omeprazole and lansoprazole protect human gastric epithelial and endothelial cells against oxidative stress. This effect was abrogated in the presence of the heme oxygenase-1 (HO-1) inhibitor ZnBG. Exposure to either PPI resulted in a strong induction of HO-1 expression on mRNA and protein level, and led to an increased activity of this enzyme. Expression of cyclooxygenase isoforms 1 and 2 remained unaffected, and COX-inhibitors did not antagonize HO-1 induction by PPIs. Our results suggest that the antioxidant defense protein HO-1 is a target of PPIs in both endothelial and gastric epithelial cells. HO-1 induction might account for the gastroprotective effects of PPIs independently of acid inhibition, especially in NSAID gastropathy. Moreover, our findings provide additional perspectives for a possible but yet unexplored use of PPIs in vasoprotection. PMID- 16712796 TI - A missense mutation in a ubiquitously expressed protein, vinculin, confers susceptibility to hypertrophic cardiomyopathy. AB - The R975W mutation, in the alternatively spliced exon 19 of vinculin (VCL) which yields the isoform metavinculin, was associated previously with hypertrophic cardiomyopathy (HCM) and dilated cardiomyopathy (DCM), and shown to alter in vivo organization of intercalated discs. We tested the hypothesis that alterations in the ubiquitously expressed, VCL-encoded protein, vinculin, may provide a pathogenic substrate for HCM. Comprehensive mutational analysis of VCL's 22 translated exons was performed in a cohort of 228 unrelated patients with genotype negative HCM, having no identifiable mutations in 12 HCM-associated myofilament/Z-disc-encoding genes. A novel missense mutation, L277M-VCL, involving a conserved residue was identified in a patient with severely obstructive, mid-ventricular hypertrophy. This mutation was not detected in 400 reference alleles. Immunohistochemical analysis of the proband's myectomy specimen demonstrated markedly reduced vinculin levels in the intercalated discs. We provide the first report of a cardiomyopathy associated mutation in vinculin. Despite its ubiquitous expression, the HCM-associated VCL mutation clinically yielded a cardiac-specific phenotype. PMID- 16712797 TI - Hyperlipidemia is a major determinant of neointimal formation in LDL receptor deficient mice. AB - LDL receptor-deficient (LDLR(-/-)) mice exhibit mild hyperlipidemia on a chow diet but develop severe hyperlipidemia on a high fat diet. In this study, we investigated neointimal formation after removal of the endothelium when LDLR(-/-) mice were fed chow or a Western diet containing 42% fat, 0.15% cholesterol, and 19.5% casein. At 10 weeks of age, female mice underwent endothelial denudation of the left common carotid artery. Two weeks after injury, neointimal formation was barely detectable in the injured vessel when mice developed mild hyperlipidemia on the chow diet. In contrast, neointimal lesions were obvious when mice developed severe hyperlipidemia on the Western diet. Immunohistochemical and histological analyses demonstrated the presence of macrophage foam cells and smooth muscle cells in neointimal lesions. The injured artery also exhibited a significant increase in medial area on the Western diet. Plasma levels of MCP-1 and soluble VCAM-1 were significantly elevated by feeding of the Western diet. These data indicate that hyperlipidemia aggravates neointimal growth in LDLR(-/-) mice by promoting foam cell formation and inflammation. PMID- 16712791 TI - Identification of intrahepatic cholangiocarcinoma related genes by comparison with normal liver tissues using expressed sequence tags. AB - Intrahepatic cholangiocarcinoma (ICC), a malignant tumor derived from the bile duct epithelium, is one of the leading causes of death from cancer, worldwide. However, the mechanisms related to it remain largely unknown. In this study, an analysis of the gene expression profiles for ICC was done using the frequency of the ESTs obtained from nine cDNA libraries that constructed from 4 ICC cell lines and 4 normal liver tissues. One hundred and thirty-seven genes were identified as being either up- or down-regulated in human ICC cells. Thirty genes were randomly selected to confirm their differential expression in 4 human ICC cell lines and 5 ICC tissues compared to normal liver tissues by semi-quantitative RT-PCR. Among these genes, ANXA1, ANXA2, AMBP, and SERPINC1 were further verified by immunohistochemical analyses. In conclusion, these identified genes represent potential biomarkers for ICC and represent potential targets for elucidating the molecular mechanisms that are associated with ICC. PMID- 16712798 TI - Sorting nexin 17, a non-self-assembling and a PtdIns(3)P high class affinity protein, interacts with the cerebral cavernous malformation related protein KRIT1. AB - The mammalian sorting nexin (SNX) proteins are involved in the endocytosis and the sorting machinery of transmembrane proteins. Additionally to the family defining phox homology (PX) domain, SNX17 is the only member with a truncated FERM (4.1, ezrin, radixin, and moesin) domain and a unique C-terminal region (together designated as FC unit). By gel filtration and lipid overlay assays we show that SNX17 is a non-self-assembling and a PtdIns(3)P high class affinity protein. A SNX17 affinity to any other phosphoinositides was not detected. By yeast two-hybrid- and GST-trapping assays we identified KRIT1 (krev1 interaction trapped 1) as a new specific interaction partner of the FC unit of SNX17. KRIT1 binds SNX17 by its N-terminal region like the known interaction partner ICAP1alpha (integrin cytoplasmic domain-associated protein-1). The interaction was also detected in HEK 293 cells transiently expressing GFP-tagged KRIT1 and Xpress-tagged SNX17. KRIT1 mutations cause cerebral cavernous malformation (CCM1). Our finding suggests a SNX17 involvement in the indicated KRIT1 function in cell adhesion processes by integrin signaling. PMID- 16712799 TI - Leucovorin-induced resistance against FDH growth suppressor effects occurs through DHFR up-regulation. AB - 10-Formyltetrahydrofolate dehydrogenase (FDH) converts 10-formyltetrahydrofolate to tetrahydrofolate (THF). Expression of the enzyme in FDH-deficient cancer cells induces cytotoxicity that can be reversed by supplementation with high concentrations of a reduced folate, 5-formyl-THF (leucovorin). In contrast, non tumor cells are resistant to FDH. The present study was undertaken to investigate mechanisms that could protect cells against FDH suppressor effects. Using 10 microM leucovorin supplementation of FDH-sensitive A549 cells transfected for FDH expression, we selected clones that have acquired resistance against FDH. Resistant cells expressed high levels of FDH and were capable of growing after withdrawal of leucovorin. These cells, however, have increased doubling time due to prolonged S phase. They also have significantly increased levels of total folate pool and THF/5,10-methylene-THF pool while the level of 10-formyl-THF was two-fold lower than in parental FDH-sensitive cells. We have shown that the FDH catalyzed reaction proceeds at about a three-fold slower rate at the ratio of 10 formyl-THF/THF corresponding to the resistant cells than at the ratio corresponding to parental sensitive cells, due to product inhibition (KI is 2.35 microM). FDH-resistant cells have strongly up-regulated dihydrofolate reductase (DHFR) that is proposed to be a mechanism for the alteration of folate pools and a key component of the acquired resistance. Elevation of DHFR in A549 cells by transient transfection decreased sensitivity to FDH toxicity and allowed selection of FDH-resistant clones. DHFR-induced repression of FDH catalysis could be an S phase-related metabolic adjustment that provides protection against FDH suppressor effects. PMID- 16712800 TI - RETRACTED: Glutathione as a mediator of apoptotic cell signaling pathways. AB - This article has been retracted consistent with Elsevier Policy on Article Withdrawal. Please see . The Publisher apologises for any inconvenience this may cause. PMID- 16712801 TI - Prefrontal mechanisms in extinction of conditioned fear. AB - Interest in the medial prefrontal cortex (mPFC) as a source of behavioral inhibition has increased with the mounting evidence for a functional role of the mPFC in extinction of conditioned fear. In fear extinction, a tone-conditioned stimulus (CS) previously paired with a footshock is presented repeatedly in the absence of footshock, causing fear responses to diminish. Here, we review converging evidence from different laboratories implicating the mPFC in memory circuits for fear extinction: (1) lesions of mPFC impair recall of extinction under various conditions, (2) extinction potentiates mPFC physiological responses to the CS, (3) mPFC potentiation is correlated with extinction behavior, and (4) stimulation of mPFC strengthens extinction memory. These findings support Pavlov's original notion that extinction is new learning, rather than erasure of conditioning. In people suffering from posttraumatic stress disorder (PTSD), homologous areas of ventral mPFC show morphological and functional abnormalities, suggesting that extinction circuits are compromised in PTSD. Strategies for augmenting prefrontal function for clinical benefit are discussed. PMID- 16712802 TI - Psychopathology in females with attention-deficit/hyperactivity disorder: a controlled, five-year prospective study. AB - BACKGROUND: Despite the importance of understanding the long-term outcome of children with attention-deficit/hyperactivity disorder (ADHD), the available literature is predominantly based on male samples. This study estimated the lifetime burden of comorbid psychopathology in a large sample of girls with and without ADHD followed up over five years. METHODS: We conducted a blind, five year prospective longitudinal study of girls with (n=140) and without (n=122) ADHD, aged 6-18 years at baseline, consecutively ascertained from either community pediatricians or psychiatrists at an academic medical center. At the five-year follow-up, 123 (88%) and 112 (92%) of the ADHD and control children, respectively, were re-assessed at a mean age of 16.7 years. Psychiatric disorders were assessed using blinded structured diagnostic interviews. RESULTS: At follow up, females with ADHD were at significantly higher risk than controls to manifest disruptive behavior, mood and anxiety disorders, and substance dependence. The magnitude of increased risk was greatest for major depression and oppositional defiant disorder, followed by substance dependence and anxiety disorders. CONCLUSIONS: These prospective follow-up findings documenting high morbidity associated with ADHD extend to females previously reported findings in male samples and underscore the importance of early recognition and intervention efforts for youth with ADHD of both genders. PMID- 16712803 TI - Involvement of Na(+), K(+)-ATPase and endogenous digitalis-like compounds in depressive disorders. AB - BACKGROUND: Sodium and potassium-activated adenosine triphosphatase (Na(+), K(+) ATPase) and endogenous digitalis-like compounds (DLC) in the brain have been implicated in the pathogenesis of mood disorders. This hypothesis was examined by the determination of Na(+), K(+)-ATPase/DLC system in parietal cortex of patients with different mood disorders and two animal models of depression. METHODS: Na(+), K(+)-ATPase concentrations in human brain synaptosomal fractions, from patients with mood disorders, schizophrenia, and normal individuals, were determined by (3)H-ouabain binding assay. Alpha isoforms were quantified by Western blotting. Brain DLC were measured using sensitive enzyme linked immunosorbant assay (ELISA). The effects of ouabain and ouabain-antibodies on behavior were determined in two animal models of depression. RESULTS: (3)H ouabain binding in bipolar patients was significantly lower than in major depressed and schizophrenic patients. Na(+), K(+)-ATPase alpha isoforms in synaptosomal fractions were not different among the groups. DLC levels in the parietal cortex of bipolar patients were significantly higher than in normal individuals and depressed patients. Injection of lipopolysaccharide (intraperitoneally) to rats elicited depression-like symptoms, which were significantly attenuated by pre-injection of ouabain-antibodies. Injection of ouabain and ouabain-antibodies (intracerebroventricular) reduced depression-like symptoms in the forced swimming test in rats. CONCLUSIONS: The results support the possibility that Na(+), K(+)-ATPase and endogenous DLC participate in the pathogenesis of depressive disorders. PMID- 16712804 TI - Activation in ventral prefrontal cortex is sensitive to genetic vulnerability for attention-deficit hyperactivity disorder. AB - BACKGROUND: Attention-deficit hyperactivity disorder (ADHD) is a heritable neuropsychiatric disorder, associated with atypical patterns of brain activation in functional imaging studies. Neuroimaging measures may serve as an intermediate phenotype in genetic studies of ADHD, as they are putatively more closely linked to gene expression than a clinical diagnosis. METHODS: We used rapid, mixed trial, event-related functional magnetic resonance imaging (fMRI) to investigate changes in brain activation during a go no-go task in boys with ADHD, their unaffected siblings, and matched control subjects. RESULTS: On the hardest inhibitory trials in our task, children and adolescents with ADHD had lower accuracy than control subjects, whereas their unaffected siblings did not. Control subjects activated a network of regions, including ventral prefrontal and inferior parietal cortex. Both children and adolescents with ADHD and their unaffected siblings showed decreased activation in these areas, as well as fewer correlations between performance and activation. CONCLUSIONS: These findings suggest that the magnitude of activation during successful inhibitions is sensitive to genetic vulnerability for ADHD in a number of regions, including ventral prefrontal cortex. If this can be replicated in future studies, this suggests that neuroimaging measures related to inhibitory control may be suitable as intermediate phenotypes in studies investigating gene effects in ADHD. PMID- 16712805 TI - Relationship of early life stress and psychological functioning to adult C reactive protein in the coronary artery risk development in young adults study. AB - BACKGROUND: Low socioeconomic status (SES) and a harsh family environment in childhood have been linked to mental and physical health disorders in adulthood. The objective of the present investigation was to evaluate a developmental model of pathways that may help explain these links and to relate them to C-reactive protein (CRP) in the Coronary Artery Risk Development in Young Adults (CARDIA) dataset. METHODS: Participants (n = 3248) in the CARDIA study, age 32 to 47 years, completed measures of childhood SES (CSES), early family environment (risky families [RF]), adult psychosocial functioning (PsyF, a latent factor measured by depression, mastery, and positive and negative social contacts), body mass index (BMI), and C-reactive protein. RESULTS: Structural equation modeling indicated that CSES and RF are associated with C-reactive protein via their association with PsyF (standardized path coefficients: CSES to RF, RF to PsyF, PsyF to CRP, CSES to CRP, all p < .05), with good overall model fit. The association between PsyF and CRP was partially mediated by BMI (PsyF to BMI, BMI to CRP, both p < .05). CONCLUSIONS: Low childhood SES and a harsh early family environment appear to be related to elevated C-reactive protein in adulthood through pathways involving psychosocial dysfunction and high body mass index. PMID- 16712806 TI - Protective effects of minocycline on the reduction of dopamine transporters in the striatum after administration of methamphetamine: a positron emission tomography study in conscious monkeys. AB - BACKGROUND: Positron emission tomography (PET) studies of methamphetamine (METH) abusers suggest that psychotic symptoms of METH abusers may be attributable to the reduction of dopamine transporters (DAT) in the human brain. However, there are currently no particular pharmacological treatments for the wide range of symptoms associated with METH abuse. METHODS: Using a PET study in conscious monkeys, we investigated whether the second generation antibiotic minocycline could protect against the reduction of DAT in monkeys treated with METH (2 mg/kg x 3, 3-hour intervals). RESULTS: Pretreatment and subsequent administration of minocycline significantly attenuated the reduction of DAT in the striatum of monkeys treated with METH. Furthermore, posttreatment and subsequent administration of minocycline also significantly attenuated the reduction of DAT. In contrast, repeated administration of minocycline alone did not alter the density of DAT in the striatum of monkeys treated with METH. CONCLUSIONS: Our findings suggest that minocycline protects against METH-induced neurotoxicity in the monkey brain. Therefore, minocycline is likely to be a promising therapeutic agent for the treatment of several symptoms associated with METH use in humans. PMID- 16712807 TI - Local knockdown of genes in the brain using small interfering RNA: a phenotypic comparison with knockout animals. AB - BACKGROUND: Recent reports have suggested effectiveness of RNA interference (RNAi) for the analysis of gene functions in the brain. This study sought to determine the efficiency of local small interfering RNA (siRNA) injections, comparing this approach with animals generated through classical gene targeting. METHODS: Small interfering RNA against dopamine transporter (DAT) (35 microg/14 days) or tyrosine hydroxylase (TH) (15 microg/3 days) was injected into the ventral tegmental/substantia nigra areas of the brain of adult wildtype or DAT knockout mice, respectively. RESULTS: Local injections of siRNA resulted in a 35% to 40% reduction of DAT and TH protein levels in the striatum, respectively. Despite negligible effect of DAT knockdown on novelty-induced locomotion, the locomotor response of DAT siRNA treated animals to amphetamine was blunted similar to what is observed in the DAT heterozygote animals. Since incomplete reduction of TH levels in normal mice does not produce behavioral effects, TH siRNA experiments were carried out in DAT-knockout animals that show increased dependence on newly synthesized dopamine. Knockdown of TH in these animals resulted in reduced basal locomotion. CONCLUSIONS: Local injection of siRNA in the brain reduced gene expression by 40% to 50%, suggesting that siRNA-mediated knockdown of genes in the brain can be a complementary tool to classical transgenesis for the analysis of gene functions. PMID- 16712809 TI - Behavioral and physiological effects of a single injection of rat interferon alpha on male Sprague-Dawley rats: a long-term evaluation. AB - Interferon-alpha (IFN-alpha) is a cytokine used as a first line of defense against diseases such as cancer and hepatitis C. However, reports indicate that its effectiveness as a treatment is countered by central nervous system (CNS) disruptions in patients. Our work explored the possibility that it may also cause long-term behavioral disruptions by chronicling the behavioral and physiological disturbances associated with a single injection of vehicle, 10, 100, or 1,000 units of IFN-alpha in male Sprague-Dawley rats (n = 5/dose). Following 1 day of locomotor baseline collection, we monitored sickness behaviors (ptosis, piloerection, lethargy, and sleep), food and water intake, body weight, temperature, and motor activity. Observations were recorded 4 days prior to and 4 days following the IFN-alpha injection. Temperature and sickness behaviors were recorded three times daily at 9:00, 15:00, and 21:00 h, and all other indices, once daily. On the injection day, temperature values were highest in the animals receiving the 10-unit IFN-alpha dose 15 min and 13 h post-injection. In the case of sickness behaviors, a significant increase was observed in piloerection in all IFN-alpha groups at each time point measured, while the scores of the rats in the vehicle condition remained unchanged between pre- and post-injection days. Analyses of overall sickness behaviors during morning and night observation periods indicated increased scores in all IFN-alpha groups following injection. Cumulatively, these data suggest that a single IFN-alpha exposure may elicit long term behavioral disruptions and that its consequences should be thoroughly investigated for its use in clinical populations. PMID- 16712808 TI - Tachyphylaxis to 5-HT3-receptor-mediated activation of vagal afferents is prevented by co-activation of 5-HT2 receptors. AB - Functional studies have provided evidence that 5-HT(3) ion-channel receptors (5 HT(3)Rs) on vagal cardiopulmonary afferents mediating the Bezold-Jarisch reflex (BJR) rapidly desensitize upon repeated exposure to selective 5-HT(3)R agonists. G-protein-coupled 5-HT(2) receptors (5-HT(2)Rs) also exist on vagal afferents, although activation of these receptors does not elicit the BJR. However, there is in vivo evidence that 5-HT(2)Rs may regulate the activity of 5-HT(3)Rs. The aim of this study was to determine whether co-activation of 5-HT(2)Rs prevents desensitization of 5-HT(3)Rs mediating the BJR in conscious rats. The principal findings were that (1) tachyphylaxis rapidly developed to the BJR-mediated hemodynamic responses elicited by successive injections of 5-HT(3)R agonists and (2) co-injection of the selective 5-HT(2)R agonist, alpha-methyl-5-HT, prevented tachyphylaxis to the BJR-mediated hemodynamic responses elicited by the 5-HT(3)R agonists. Additional studies provided evidence that (1) tachyphylaxis to the 5 HT(3)R agonists was not due to impairment of the central or efferent processing of the BJR, and (2) the pressor responses elicited by alpha-methyl-5-HT were not responsible for preventing tachyphylaxis to the BJR reflex responses elicited by 5-HT(3)R agonists. These results suggest that the loss of response to 5-HT(3)R agonists is due to desensitization of 5-HT(3)Rs on vagal afferents mediating the BJR and that co-activation of 5-HT(2)Rs prevents the desensitization of these 5 HT(3)Rs. PMID- 16712810 TI - Does the error negativity reflect the degree of response conflict? AB - The recruitment of performance control was recently explained in terms of the detection of response conflict. One recent hypothesis claims that the degree of response conflict is reflected in an electroencephalographical component, the Error Negativity (Ne). Indeed, in the framework of simulations, variations of the Ne fitted well with variations of response conflict. The objective of the present study was to test the conflict hypothesis of the Ne in a real experimental situation. The subjects performed an arrowhead flanker task with two alternative responses. The degree of conflict was measured by the degree of the co-activation of the forces exerted by the two alternative effectors. Trials with co activations (i.e., an incorrect response activation followed by a correct one) were selected. These trials were divided into two categories, namely trials with suprathreshold error force (full errors) and subthreshold error force (partial errors). The results showed that although the degree of conflict was larger for full than for partial errors, the Ne was not different for the two categories. Our study does not support the hypothesis that the Ne amplitude reflects the degree of conflict. PMID- 16712811 TI - Upregulation of calbindin-D28k expression during regeneration in the adult fish cerebellum. AB - In contrast to mammals, fish are distinguished by their enormous potential for brain repair after injuries. This phenomenon has been well studied after application of stab-wound lesions to the corpus cerebelli, a cerebellar subdivision, in the teleost fish Apteronotus leptorhynchus. By combining this lesion paradigm with immunohistochemical staining, we examined the potential role of the calcium-binding protein calbindin-D(28k) in the process of regeneration. Calbindin-D(28k)-immunoreactive cell bodies and fibers were evident in the lesion path and the immediate vicinity of the lesion in the period between 16 h and 7 days after the lesion but absent from this region at shorter or longer postlesion survival times and in the intact brain. Both the number of immunolabeled cells and the intensity of the label were most pronounced 1-3 days postlesion. Analysis of the morphology of the immunostained cells by confocal microscopy suggested that most, and perhaps all of them, were granular neurons. Since the transient upregulation of calbindin-D(28k) is paralleled by a decline in the number of cells undergoing apoptotic cell death, we hypothesize that this protein exerts a neuroprotective function, probably by buffering free intracellular Ca(2+), whose concentration is elevated after brain insults. PMID- 16712812 TI - A histopathological study of premature and mature infants with pontosubicular neuron necrosis: neuronal cell death in perinatal brain damage. AB - Perinatal hypoxic-ischemic brain damage is a major cause of neuronal and behavior deficits, in which the onset of injury can be before, at or after birth, and the effects may be delayed. Pontosubicular neuron necrosis (PSN) is one of perinatal hypoxic-ischemic brain injury and its pathological peculiarity is neuronal apoptosis. In this study, we investigated whether apoptotic cascade of PSN used a caspase-pathway or not, and whether hypoglycemia activated apoptosis or not. Sections of the pons of PSN with and without hypoglycemia were stained using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) and immunohistochemistry for glial fibrillary acidic protein (GFAP), Bcl-2, Bcl-x and activated caspase 3. Additionally, we performed immunoblot analysis of Bcl-2, Bcl-x and activated caspase 3. TUNEL-positive cell was closely associated with the presence of karyorrhexis. Under combination of karyorrhectic and TUNEL-positive cells, number of apoptotic cells in premature brains was significantly more than in mature brains. Hypoxic-ischemic brain injury was considered to easily lead to apoptosis in premature infants. Moreover, as this pathophysiology, caspase-pathway activation contributed to neuronal death from caspase-immunoexpression analyses. PSN with hypoglycemia showed large number of apoptotic cells and higher expression of activated caspase 3. The result may be more severe with the background of hypoglycemia and prematurity complicated by hypoxia and/or ischemia. PMID- 16712813 TI - Enhancement of morphine antinociception with the peptide N-methyl-D-aspartate receptor antagonist [Ser1]-histogranin in the rat formalin test. AB - Opiates may be used to attenuate chronic pain, but long-term use is complicated by the possible increase in pain over time, escalating dose requirements, and untoward side effects. Adjuncts such as ketamine, an N-methyl-D-aspartate (NMDA) receptor antagonist, may be added to opiates to provide more consistent analgesia. However, the unwanted motor side effects of NMDA receptor antagonists prevent their widespread clinical usage. In the current study, an analogue of the naturally-derived peptide histogranin, [Ser(1)]histogranin (SHG), an NMDA receptor modulator without adverse side effects like those in current clinical use, was evaluated for its potential to enhance the antinociceptive effect of intrathecal morphine in the rat formalin test. Intrathecal injection of a combination of SHG and morphine resulted in significantly reduced hind paw flinching compared with morphine alone in the first and second phases. The effective dose of SHG used in the combination had no efficacy when tested alone. These results were similar to the increased efficacy that was obtained with a combination of ketamine and morphine. Thus, enhancement of opiate efficacy is possible using a novel peptide NMDA receptor modulator with a potentially improved safety profile. PMID- 16712814 TI - One hour, but not six hours, of daily access to self-administered cocaine results in elevated levels of the dopamine transporter. AB - We have previously shown that brief (1 h) and extended (6 h) daily access to IV cocaine self-administration produce different behavioral and neural consequences following 2 weeks of drug withdrawal. Brief daily access produced stable consumption of the drug and, after withdrawal, a sensitized locomotor response and an enhanced c-Fos labeling to a single cocaine challenge. In contrast, extended daily cocaine self-administration produced escalation of drug consumption over trials but no enhanced behavioral or neurochemical response after withdrawal. Cocaine affects dopaminergic (DA) function by binding to the presynaptic transporter and thereby preventing reuptake of the neurotransmitter an action thought to be responsible for the drug's reinforcing properties. In an extension of our previous work, the current study, using receptor autoradiography, compared binding (by [3H]WIN35428) of the dopamine transporter (DAT) in animals having experienced either brief or extended daily access to cocaine over 8 days, followed by 14 days of withdrawal. DAT densities were found to increase in the nucleus accumbens core (N.Acc Core) and the dorsal striatum (but not in the N.Acc shell, medial prefrontal cortex (mPFC), or ventral tegmental area (VTA)) of the 1-h, but not 6-h, subjects. In other words, elevations in DAT density were not associated with the 6-h access group, the group that models patterns of drug-use in human addicts, and therefore are likely to be independent of the neuroadaptations that occur in the "addictive" process. Such conclusions are also consistent with brain-imaging studies of human cocaine addicts. Additional research will be needed to identify the specific neural changes relevant to addiction. PMID- 16712816 TI - The perception of musical phrase structure: a cross-cultural ERP study. AB - Electroencephalography (EEG) was used in a cross-cultural music study investigating phrase boundary perception. Chinese and German musicians performed a cultural categorization task under Chinese and Western music listening conditions. Western music was the major subject for both groups of musicians, while Chinese music was familiar to Chinese subjects only. By manipulating the presence of pauses between two phrases in the biphrasal melodies, EEG correlates for the perception of phrase boundaries were found in both groups under both music listening conditions. Between 450 and 600 ms, the music CPS (closure positive shift), which had been found in earlier studies with a false tone detection task, was replicated for the more global categorization task and for all combinations of subject group and musical style. At short latencies (100 and 450 ms post phrase boundary offset), EEG correlates varied as a function of musical styles and subject group. Both bottom-up (style properties of the music) and top-down (acculturation of the subjects) information interacted during this early processing stage. PMID- 16712815 TI - Rapid discrimination of visual scene content in the human brain. AB - The rapid evaluation of complex visual environments is critical for an organism's adaptation and survival. Previous studies have shown that emotionally significant visual scenes, both pleasant and unpleasant, elicit a larger late positive wave in the event-related brain potential (ERP) than emotionally neutral pictures. The purpose of the present study was to examine whether neuroelectric responses elicited by complex pictures discriminate between specific, biologically relevant contents of the visual scene and to determine how early in the picture processing this discrimination occurs. Subjects (n = 264) viewed 55 color slides differing in both scene content and emotional significance. No categorical judgments or responses were required. Consistent with previous studies, we found that emotionally arousing pictures, regardless of their content, produce a larger late positive wave than neutral pictures. However, when pictures were further categorized by content, anterior ERP components in a time window between 200 and 600 ms following stimulus onset showed a high selectivity for pictures with erotic content compared to other pictures regardless of their emotional valence (pleasant, neutral, and unpleasant) or emotional arousal. The divergence of ERPs elicited by erotic and non-erotic contents started at 185 ms post-stimulus in the fronto-central midline region, with a later onset in parietal regions. This rapid, selective, and content-specific processing of erotic materials and its dissociation from other pictures (including emotionally positive pictures) suggests the existence of a specialized neural network for prioritized processing of a distinct category of biologically relevant stimuli with high adaptive and evolutionary significance. PMID- 16712817 TI - Dissociated cortical networks show spontaneously correlated activity patterns during in vitro development. AB - In vitro cultured neuronal networks coupled to microelectrode arrays (MEAs) constitute a valuable experimental model for studying changes in the neuronal dynamics at different stages of development. After a few days in culture, neurons start to connect each other with functionally active synapses, forming a random network and displaying spontaneous electrophysiological activity. The patterns of collective rhythmic activity change in time spontaneously during in vitro development. Such activity-dependent modifications play a key role in the maturation of the network and reflect changes in the synaptic efficacy, fact widely recognized as a cellular basis of learning, memory and developmental plasticity. Getting advantage from the possibilities offered by the MEAs, the aim of our study is to analyze and characterize the natural changes in dynamics of the electrophysiological activity at different ages of the culture, identifying peculiar steps of the spontaneous evolution of the network. The main finding is that between the second and the third week of culture, the network completely changes its electrophysiological patterns, both in terms of spiking and bursting activity and in terms of cross-correlation between pairs of active channels. Then the maturation process can be characterized by two main phases: modulation and shaping in the synaptic functional connectivity of the network (within the first and second week) and general moderate correlated activity, spread over the entire network, with connections properly formed and stabilized (within the fourth and fifth week). PMID- 16712818 TI - Aspirin protects striatal dopaminergic neurons from neurotoxin-induced degeneration: an in vivo microdialysis study. AB - The effect of aspirin on dopaminergic neuronal damage induced by in vivo infusion of 1-methyl-4-phenylpiridinium iodide (MPP(+)) and 6-hydroxydopamine (6-OHDA) was studied in rats, using microdialysis. Rat striata were perfused with 1 mM MPP(+) or 6-OHDA for 10 min, causing peak levels of dopamine (DA) in the dialytic fluid, after 40 min. After 24 h, 1 mM MPP(+) was perfused again for 10 min and DA levels measured in the dialytic fluid, as an index of neuronal cell integrity. Pretreatment with Aspidol (lysine acetylsalicylate), 180 mg/kg i.p., 1 h before MPP(+) or 6-OHDA perfusion, did not modify DA extracellular output, on day 1, but restored MPP(+)-induced DA release on day 2, indicating a neuroprotective effect of Aspidol. Conversion of 0.5 mM 4-hydroxybenzoic acid (4-HBA) to 3,4 dihydroxybenzoic acid (3,4-DHBA) was measured as an index of reactive oxygen species (ROS). 6-OHDA, but not MPP(+), significantly enhanced 3,4-DHBA levels in the perfusion fluid. Aspidol (180 mg/kg, i.p.) reduced 6-OHDA-dependent increase of 3,4-DHBA levels. Meloxicam (50 mg/kg, i.p.), a specific cyclooxygenase-2 (COX 2) inhibitor, was ineffective against both neurotoxins. These data suggest that the protective effect of aspirin is due to different mechanisms of action according to the neurotoxin used, and it is independent from COX-2 inhibition. PMID- 16712819 TI - Behavioral and morphological effects of minocycline in the 6-hydroxydopamine rat model of Parkinson's disease. AB - The neuropathology in many neurodegenerative diseases is mediated by inflammatory cascades that influence neuronal dysfunction and death. Minocycline reduces the neurodegeneration observed in various models of Parkinson's. We exploited the unilateral 6-hydroxydopamine (6-OHDA) lesion model to assess the effect of minocycline on related neurodegeneration. Thirty Fisher 344 rats were divided into three daily treatment groups: (1) after: 45 mg/kg of minocycline beginning 24 h after lesioning; (2) before: 45 mg/kg of minocycline beginning 3 days before 6-OHDA lesioning; (3) control: corresponding saline-treated controls. Animals were assessed for apomorphine-induced rotations for 4 weeks. A longitudinal model for repeated measures showed that both after and before groups had significantly lower rotations than controls (P < 0.001 for both comparisons). Pair-wise group comparisons showed that the before animals rotated less compared to controls (mean rotations: 164 +/- 38 versus 386 +/- 49, respectively, P = 0.001). After animals also rotated significantly less then controls (mean rotations: 125 +/- 41 versus 386 +/- 49, respectively, P < 0.001). Animals receiving minocycline displayed reduced tyrosine hydroxylase-positive cell loss in the lesioned nigra versus contralateral nonlesioned nigra, compared to controls (mean differences: 5065 for after, 3550 for before, and 6483 for controls; P = 0.158 for after versus controls, P = 0.019 for before versus controls). The remaining lesioned nigral cells of both minocycline-treated groups were larger than controls, with the most robust cell size and fiber density observed in the after group. These data suggest that the therapeutic potential of minocycline may depend on the time of drug administration relative to neuropathogenic event. PMID- 16712820 TI - Oxidation and cytotoxicity of 6-OHDA are mediated by reactive intermediates of COX-2 overexpressed in PC12 cells. AB - Parkinson's disease is characterized by a progressive loss of dopaminergic neurons, likely associated with dysregulation of oxidation of catechols, such as dopamine (DA) and 6-hydroxydopamine (6-OHDA), and resulting in oxidative stress. The involvement of cyclooxygenase-2 (COX-2) in pathogenesis of Parkinson's disease has been suggested. However, specific COX-2 triggered mechanisms participating in catalysis of DA oxidation and enhanced catechol-induced cytotoxicity remain poorly characterized. Here, we demonstrate that in a model biochemical system, recombinant heme-reconstituted COX-2 induced oxidation of 6 OHDA in the course of its peroxidase (H(2)O(2)-dependent) and cyclooxygenase (arachidonic acid (AA)-dependent) catalytic half-cycles. Similarly, COX-2 was able to stimulate 6-OHDA oxidation during its peroxidase- and cyclooxygenase half cycles and caused oxidative stress in homogenates of PC12 cells stably overexpressing the enzyme (but not in mock-transfected cells). In addition, the increased levels of COX-2 were associated with enhanced cytotoxicity of 6-OHDA in stably transfected PC12 cells. Finally, co-oxidation of 6-OHDA by COX-2 triggered production of superoxide radicals critical for both propagation of 6-OHDA oxidation and induction of oxidative stress in COX-2 overexpressing cells. Thus, we conclude that both peroxidase and cyclooxygenase half-cycles of COX-2 catalyzed reactions are essential for COX-2-dependent activation of 6-OHDA oxidation, oxygen radical production, oxidative stress, and cytotoxicity. PMID- 16712821 TI - Preparation of the pentasaccharide hapten of the GPL of Mycobacterium avium serovar 19 by achieving the glycosylation of a tertiary hydroxyl group. AB - The chemical synthesis of the glycopeptidolipid-type pentasaccharide hapten of Mycobacterium avium serovar 19 with a trifluoroacetamido spacer at the reducing end is described. The spacer-armed pentasaccharide 31, when conjugated to an immunogenic protein, can be applied to the serodiagnosis of mycobacterial infections. The questionable structure of the penultimate monosaccharide unit was clarified as 6-deoxy-3-C-methyl-2,4-di-O-methyl-L-mannopyranose. The occurrence of the 6-deoxy-3-C-methyl-2,4-di-O-methyl-L-talopyranose could be excluded by the presence of the large H-1'-H-2' coupling constant, which proves the 4C1 (L) conformation as the favoured one. PMID- 16712822 TI - Semi-synthetic heparin derivatives: chemical modifications of heparin beyond chain length, sulfate substitution pattern and N-sulfo/N-acetyl groups. AB - The glycosaminoglycan heparin is a polyanionic polysaccharide most recognized for its anticoagulant activity. Heparin binds to cationic regions in hundreds of prokaryotic and eukaryotic proteins, termed heparin-binding proteins. The endogenous ligand for many of these heparin-binding proteins is a structurally similar glycosaminoglycan, heparan sulfate (HS). Chemical and biosynthetic modifications of heparin and HS have been employed to discern specific sequences and charge-substitution patterns required for these polysaccharides to bind specific proteins, with the goal of understanding structural requirements for protein binding well enough to elucidate the function of the saccharide-protein interactions and/or to develop new or improved heparin-based pharmaceuticals. The most common modifications to heparin structure have been alteration of sulfate substitution patterns, carboxyl reduction, replacement N-sulfo groups with N acetyl groups, and chain fragmentation. However, an accumulation of reports over the past 50 years describe semi-synthetic heparin derivatives obtained by incorporating aliphatic, aryl, and heteroaryl moieties into the heparin structure. A primary goal in many of these reports has been to identify heparin derived structures as new or improved heparin-based therapeutics. Presented here is a perspective on the introduction of non-anionic structural motifs into heparin structure, with a focus on such modifications as a strategy to generate novel reduced-charge heparin-based bind-and-block antagonists of HS-protein interactions. The chemical methods employed to synthesize such derivatives, as well as other unique heparin conjugates, are reviewed. PMID- 16712823 TI - Antigen-specific accumulation of naive, memory and effector CD4 T cells during anterior uveitis monitored by intravital microscopy. AB - Uveitis is an immune-mediated ocular disease and a leading cause of blindness. We characterized a novel model of uveitis with intravital microscopy. Transfer of ovalbumin-specific T cells from DO11.10 spleen to BALB/c recipients and subsequent challenge with ovalbumin in the anterior chamber of the eye resulted in anterior uveitis. Antigen-specificity was verified by injection of irrelevant antigen and transfer of T cells with a different specificity. Subsets of CD4 T cells, including naive (DO11.10 RAG(-/-)) and in vitro-activated Th2 effector CD4 T cells, infiltrated anterior segment tissues early in the inflammation. Memory like CD44(high) CD4 T cells from unprimed transgenic mice and in vitro-activated Th1 effector CD4 T cells accumulated to larger numbers than naive or Th2 effector cells at 48 and 72 h. Of these, the alpha(2)-integrin+CD4 unprimed T cells entered the eye more efficiently, and antibody to alpha(2)-integrin markedly inhibited the inflammatory response. Intravital microscopy revealed the early arrival and antigen-specific accumulation of CD4 T cells in inflamed tissue and should be helpful in understanding T cell migration to other organs. PMID- 16712824 TI - Determination of adiponectin in serum using a latex particle-enhanced turbidimetric immunoassay with an automated analyzer. AB - BACKGROUND: Adiponectin is an adipose-derived hormone that plays a role in regulating metabolic processes such as fat partitioning and lipid and glucose metabolism. Quantification of adiponectin is useful for obtaining information on metabolic syndrome, but there is no rapid method to measure adiponectin for clinical use. METHODS: We developed a rapid and sensitive latex particle-enhanced turbidimetric immunoassay (LTIA) using a latex bead-immobilized anti-adiponectin polyclonal antibody. The assay was performed on a Hitachi H7170 analyzer and evaluated for validity as a method to quantitate adiponectin, in parallel with the ELISA. RESULTS: Dilution tests using LTIA showed linearity from 0.25 to 30 microg/ml. Within-run CV and total CV were obtained in the range of 0.8-1.9% and 1.1-2.0%, respectively. No interference was observed in the testing of specimens containing potentially interfering substances such as bilirubin, ditaurobilirubin, hemoglobin triglyceride, rheumatoid factor, type IV collagen, fibronectin, and complement factor (C1q). A strong correlation between LTIA and ELISA was confirmed (n=30, r=0.990, y=0.95x+0.39). CONCLUSION: The LTIA assay is applicable to quantitating the serum concentration of adiponectin. This assay is more convenient and faster than ELISA and suitable for clinical routine analysis. PMID- 16712825 TI - Thiol compounds metabolism in mice, rats and humans: comparative study and potential explanation of rodents protection against vascular diseases. AB - BACKGROUND: Rodents are often used as animal models to dissect mechanisms underlying hyperhomocysteinemia atherogenicity in humans. However, neither wild type rodents nor cystathionine beta-synthase deficient mice develop spontaneous atherosclerosis. We investigated whether species-specific differences in thiols metabolism may explain the respective sensitivity of rodents and humans to hyperhomocysteinemia. METHODS: Thiols and vitamins B levels were determined in normohomocysteinemic humans and rodents, and in hyperhomocysteinemic mice. RESULTS: In basal status, although plasma homocysteine, cysteine and cysteinylglycine levels were lower, glutathione levels were higher in mice than in humans (4.0+/-1.6 vs. 7.9+/-2.2, P<0.0005; 147.4+/-40.3 vs. 278.5+/-50.0, P<0.0001; 2.3+/-0.7 vs. 36.6+/-7.3, P<0.0001; and 70.9+/-20.1 vs. 4.6+/-1.6, P<0.0001). Serum vitamin B12 and folate levels were 2.5- and 7.7-fold higher in rats than in humans. In wild-type mice, the increase in plasma Hcy levels induced by methionine-enriched diet was accompanied by a proportional increase in GSH levels. CONCLUSION: GSH levels are enough to modulate Hcy effects in normo- and hyperhomocysteinemic mice but not in humans. This rodents characteristic, likely supported by species differences in the relative contribution of remethylation and transsulfuration pathways, may partly explain their protection against atherosclerosis. PMID- 16712827 TI - Twelve different enzyme assays on dried-blood filter paper samples for detection of patients with selected inherited lysosomal storage diseases. AB - BACKGROUND: Diagnoses of inherited lysosomal storage diseases are based on specific enzymatic assays performed on plasma, leukocytes, fibroblasts, and lately, dried-blood filter paper samples. We evaluated feasibility of detecting of patients with several inherited lysosomal storage diseases using dried-blood filter paper samples for appropriate enzyme assays. METHODS: Fluorometric methods were used to evaluate the activities of arylsulfatase B, alpha-N acetylglucosaminidase, chitotriosidase, alpha and beta-galactosidases, beta glucosidase, beta-glucuronidase, total hexosaminidases, hexosaminidase A, alpha iduronidase, and iduronate-2-sulfatase. A radiometric method was used for sphyngomyelinase determination. Single 3.0-mm diameter disks containing dried blood samples were incubated at 37 degrees C with appropriate dilution buffers and artificial substrates, and the fluorescence or radioactivity was measured. RESULTS: Our results showed a statistically significant difference of the enzyme activity between affected individuals and controls, in all the assays performed. In contrast, we have not obtained a complete differentiation between heterozygotes and controls with these assays. CONCLUSIONS: Enzyme assay on dried blood filter paper is a suitable method to screen for several lysosomal storage diseases. Despite the low individual incidence of these pathologies, the incorporation of individual enzyme assays in neonatal screening programs could be justified to screen for diseases with relatively high local frequency and therapeutic measures available. PMID- 16712828 TI - Cryopreservation of somatic embryos of the herbaceous peony (Paeonia lactiflora Pall.) by air drying. AB - This study was carried out to establish a suitable method for the cryopreservation of somatic embryos of the herbaceous peony. The somatic embryos were obtained from cotyledon and anther cultures on a MS medium supplemented with abscisic acid (ABA) and phenylacetic acid (PAA), respectively. The frequency of somatic embryo formation was the greatest (61%) from the cotyledons cultured on a MS medium supplemented with 1.0 mg l(-1) of ABA. Embryos were also obtained directly from anthers cultured on a MS medium with or without 2.0 mg l(-1) of PAA. For the cryopreservation of peony somatic embryos, the embryos were dried under a stream of sterile air and frozen by immersion in liquid nitrogen. Thawed embryos were germinated into plantlets after placing on a medium containing 0.3 mg l(-1) of gibberellic acid (GA(3)). The frequency of the post-thaw regrowth of cryopreserved somatic embryos was related to their size and desiccation time, the latter ranging from 0 to 2 h. When the somatic embryos were desiccated for 1 h, the frequency of post-thaw regrowth was greater than 66%. The frequency of post thaw regrowth of the cryopreserved somatic embryos from anthers and cotyledon tissues was generally high when they were 2-3 mm in size. Desiccation may be a suitable method for the cryopreservation of somatic embryos of the herbaceous peony. PMID- 16712829 TI - Sperm viability in the black-footed ferret (Mustela nigripes) is influenced by seminal and medium osmolality. AB - Fundamental knowledge of spermatozoa cryobiology can assist with optimizing cryopreservation protocols needed for genetic management of the endangered black footed ferret. Objectives were to characterize semen osmolality and assess the influence of two media at various osmolalities on sperm viability. We examined the influence of Ham's F10 +Hepes medium (H) at 270, 400, 500 or 700 mOsm (adjusted with sucrose, a nonpermeating cryoprotectant) and TEST Yolk Buffer (TYB) with 0% (300 mOsm) versus 4% (900 mOsm) glycerol (a permeating cryoprotectant). Electroejaculates (n=16) were assessed for osmolality using a vapor pressure osmometer. For media comparison, semen (n=5) was collected in TYB 0%, split into six aliquots, and diluted in H270, H400, H500, H700, and TYB 0% or TYB 4%. Each sample was centrifuged (300 g, 8 min), resuspended in respective medium, and maintained at 37 degrees C for 3h. Sperm motility and forward progression were monitored every 30 min for 3h post-washing. Acrosomal integrity was monitored at 0 and 60 min post-washing. Results demonstrated that black footed ferret semen has a comparatively high osmolality (mean+/-SEM, 513.1+/-32.6 mOsm; range, 366-791 mOsm). Ferret spermatozoa were sensitive to hyperosmotic stress. Specifically, sperm motility was more susceptible (P<0.01) to hyperosmotic conditions than acrosomal integrity, and neither were influenced (P>0.05) by hypotonic solutions. Exposure to TYB 4% glycerol retained more (P<0.01) sperm motility than a hyperosmotic Ham's (700 mOsm). These findings will guide the eventual development of assisted breeding with cryopreserved sperm contributing to genetic management of this rare species. PMID- 16712830 TI - Cryopreservation of European eel (Anguilla anguilla) spermatozoa: effect of dilution ratio, foetal bovine serum supplementation, and cryoprotectants. AB - The main aim of the present study was to investigate the effect of sperm freezing medium dilution ratio (1:1, 1:2, and 1:5 v/v), two cryoprotectants: dimethyl sulphoxide (Me(2)SO) and methanol (MeOH), and the addition of foetal bovine serum (FBS) on the cryopreservation of European eel sperm. The effect of these factors was evaluated comparing post-thawing viability with fluorescent staining (Hoechst bisbenzimide 33258) and the spermatozoa head morphometry, determined with computer-assisted morphology analysis (ASMA). The 1:5 (v/v) dilution ratio resulted in a lower viability in comparison with 1:1 and 1:2 (52.8+/-2.3% vs. 67.4+/-2.3% and 65.1+/-2.3%, respectively, p=0.0001), but without effects on the head morphology. Although the viability was not significantly different between Me(2)SO and MeOH (60.4+/-1.9 vs. 63.2+/-1.9%, respectively, p=0.305), a decrease of spermatozoa head area and perimeter was found when spermatozoa were frozen with methanol (6.19+/-0.01 vs. 6.36+/-0.01 microm(2) and 17.28+/-0.05 vs. 17.49+/ 0.05 microm, for area and perimeter and MeOH and Me(2)SO, respectively, p=0.0001). Finally, a higher viability (75.1+/-1.7 vs. 48.5+/-1.7, with or without FBS, respectively, p=0.0001) and higher spermatozoa head size (6.40+/ 0.01 vs. 6.15+/-0.01microm(2) and 17.88+/-0.05 vs. 16.89+/-0.05 microm, for area and perimeter, with or without FBS, respectively, p=0.0001) were found when cells were frozen-thawed in freezing media supplemented with FBS. Based on the above findings, dilution ratios lower than 1:5 (v/v) and the addition of serum improved the viability results after cryopreservation. Future studies are required in order to understand the spermatozoa membrane interchange mechanisms in response to the changes in spermatozoa head size caused by cryoprotectants and freezing media supplements. PMID- 16712831 TI - Progeny of germ line knockouts of ASI2, a gene encoding a putative signal transduction receptor in Tetrahymena thermophila, fail to make the transition from sexual reproduction to vegetative growth. AB - The ciliated protozoan Tetrahymena has two nuclei: a germ line micronucleus and a somatic macronucleus. The transcriptionally active macronucleus has about 50 copies of each chromosome. At sexual reproduction (conjugation), the parental macronucleus is degraded and new macronucleus develops from a mitotic product of the zygotic micronucleus. Development of the macronucleus involves massive genome remodeling, including deletion of about 6000 specific internal eliminated sequences (IES) and multiple rounds of DNA replication. A gene encoding a putative signal transduction receptor, ASI2, (anlagen stage induced 2) is up regulated during development of the new macronuclei (anlagen). Macronuclear ASI2 is nonessential for vegetative growth. Homozygous ASI2 germ line knockout cells with wild type parental macronuclei proceed through mating but arrest at late macronuclear anlagen development and die before the first post-conjugation fission. IES elimination occurs in these cells. Two rounds of postzygotic DNA replication occur normally in progeny of ASI2 germ line knockouts, but endoreduplication of the macronuclear genome is arrested. The germ line ASI2 null phenotype is rescued in a mating of a knockout strain with wild type cells. PMID- 16712832 TI - Successful ovulation in plasminogen-deficient mice treated with the broad spectrum matrix metalloproteinase inhibitor galardin. AB - Many studies have suggested the hypothesis that the plasminogen activator (PA) system and the matrix metalloproteinase (MMP) system, either separately or in combination, may provide the proteolytic activity that is required for rupture of the follicular wall at the time of ovulation. Our recent studies on ovulation in plasminogen (plg)-deficient mice have, however, shown that plasmin is not required for normal ovulation, leading us to the hypothesis that MMPs may be a more important source of proteolysis for this process. To investigate the role of MMPs and also the possibility of a functional overlap or synergy between the MMP and PA systems during ovulation, we have studied ovulation efficiency in wild type and plg-deficient mice treated with the broad-spectrum MMP inhibitor galardin. We found that in both wild-type mice and heterozygous plg-deficient (plg+/-) mice that had been treated with galardin prior to ovulation, there was a mild (18-20%) reduction in ovulation efficiency. Surprisingly, galardin treatment of plg-deficient (plg-/-) mice only caused an additional 14% reduction in ovulation efficiency as compared to vehicle-treated plg-/- mice. Our data therefore suggest that although MMPs may play a role in degradation of the follicular wall, they may not be obligatory for ovulation. In contrast to previous studies on tissue remodeling during wound healing and placental development, we have demonstrated that there is no obvious functional overlap or synergy between the PA and MMP systems, which has previously been thought to be essential for the ovulatory process. PMID- 16712833 TI - Ernest Everett Just (1883-1941)--an early ecological developmental biologist. AB - Ecological developmental biology (Eco-Devo) involves the study of development in its natural environmental context as opposed to the laboratory setting. Ernest E. Just was an early 20th century African-American embryologist who devoted his career to studying the early development of marine invertebrates in the United States and abroad. Through detailed study of the fertilization process, he came to see the cell cortex as playing a central role in development, inheritance, and evolution. This paper, after presenting some of Just's scientific and philosophical contributions, argues that Just was an Eco-Devo biologist. Three lines of evidence are given. First, Just believed that intimate knowledge of the natural history of the marine animal under study--hence, the natural setting in which fertilization occurs--was essential. Second, he stressed the importance of the egg's "normality"--how well its condition in the laboratory corresponds to the natural, fertilizable state. Finally, Just was an organicist, believing that organisms are holistic systems with emergent properties that arise from their organization and complexity. Although other scientists may stand out more clearly as founding architects of Eco-Devo, E. E. Just, with his unwavering insistence on the normality and holistic integrity of the egg cell, was one of its purest adherents. PMID- 16712834 TI - Zebrafish Trap230/Med12 is required as a coactivator for Sox9-dependent neural crest, cartilage and ear development. AB - The vertebrate Sox9 transcription factor directs the development of neural crest, otic placodes, cartilage and bone. In zebrafish, there are two Sox9 orthologs, Sox9a and Sox9b, which together perform the functions of the single-copy tetrapod Sox9. In a large-scale genetic screen, we have identified a novel zebrafish mutant that strongly resembles the Sox9a/Sox9b double mutant phenotype. We show that this mutation disrupts the zebrafish Trap230/Med12 ortholog, a member of the Mediator complex. Mediator is a coactivator complex transducing the interaction of DNA-binding transcription factors with RNA polymerase II, and our results reveal a critical function of the Trap230 subunit as a coactivator for Sox9. PMID- 16712835 TI - Multiple EGFR ligands participate in guiding migrating border cells. AB - Cell migration is an important feature of embryonic development as well as tumor metastasis. Border cells in the Drosophila ovary have emerged as a useful in vivo model for uncovering the molecular mechanisms that control many aspects of cell migration including guidance. It was previously shown that two receptor tyrosine kinases, epidermal growth factor receptor (EGFR) and PDGF- and VEGF-related receptor (PVR), together contribute to border cell migration. Whereas the ligand for PVR, PVF1, is known to guide border cells, it is unclear which of the four activating EGFR ligands function in this process. We developed an assay to detect the ability of secreted factors to reroute migrating border cells in vivo and tested the activity of EGFR ligands compared to PVF1. Two ligands, Keren and Spitz, guided border cells whereas the other ligands, Gurken and Vein, did not. In addition, only Keren and Spitz were expressed at the appropriate stage in the oocyte, the target of border cell migration. Therefore, a complex combination of EGFR and PVR ligands together guide border cells to the oocyte. PMID- 16712837 TI - Targeting cytosolic phospholipase A2 by arachidonyl trifluoromethyl ketone prevents chronic inflammation in mice. AB - Cytosolic phospholipase A(2) (cPLA(2)) plays a pivotal role in inflammation by catalyzing the release of arachidonic acid, a substrate for lipoxygenase and cyclooxygenase enzymes, from membrane phospholipids. In the present study we examined the role of cPLA(2) in inflammatory responses through the use of a specific inhibitor of the enzyme, cPLA(2), arachidonyl trifluoromethyl ketone (AACOCF3). Interestingly, we observed that AACOCF3 is an inhibitor of chronic but not acute inflammatory responses. Specifically, AACOCF3 inhibited phorbol 12 myristate 13-acetate (PMA)-induced chronic ear edema in mice. Additionally, oral treatment of ovalbumin-sensitized/ovalbumin-challenged BALB/c mice with 20 mg/kg AACOCF3 prevented the development of airway hyper-responsiveness in a model of asthma. Furthermore, AACOCF3 decreased cellular recruitment in the airway lumen and airway inflammation after the ovalbumin challenge. Taken together, these results suggest that a potent and specific chemical inhibitor of cPLA(2) may be useful for the treatment of chronic inflammatory diseases including rheumatoid arthritis, inflammatory bowel disease, psoriasis, and asthma. PMID- 16712836 TI - The BMP antagonist Noggin promotes cranial and spinal neurulation by distinct mechanisms. AB - Here we characterize the consequences of elevated bone morphogenetic protein (BMP) signaling on neural tube morphogenesis by analyzing mice lacking the BMP antagonist, Noggin. Noggin is expressed dorsally in the closing neural folds and ventrally in the notochord and somites. All Noggin-/- pups are born with lumbar spina bifida; depending on genetic background, they may also have exencephaly. The exencephaly is due to a primary failure of neurulation, resulting from a lack of mid/hindbrain dorsolateral hinge point (DLHP) formation. Thus, as previously shown for Shh signaling at spinal levels, BMP activity may inhibit cranial DLHP morphogenesis. However, the increased BMP signaling observed in the Noggin-/- dorsal neural tube is not sufficient to cause exencephaly; it appears to also depend on the action of a genetic modifier, which may act to increase dorsal Shh signaling. The spinal neural tube defect results from a different mechanism: increased BMP signaling in the mesoderm between the limb buds leads to abnormal somite differentiation and axial skeletal malformation. The resulting lack of mechanical support for the neural tube causes spina bifida. We show that this defect is due to elevated BMP4 signaling. Thus, Noggin is required for mammalian neurulation in two contexts, dependent on position along the rostrocaudal axis. PMID- 16712838 TI - Decreased association of aged alpha crystallins with gamma crystallins. AB - Previous studies have demonstrated non-covalent interactions of alpha crystallins with gamma crystallins, under true equilibrium conditions. These interactions could affect short-range interactions of lens crystallins that are necessary for the transparent properties of the lens. Since the transparent properties of the lens decrease during aging, it is possible that there are corresponding changes in the ability of aged alpha crystallins to interact with gamma crystallins. In the following study, alpha crystallins were prepared from fetal and aged bovine lenses, then tested for binding to gamma crystallins using microequilibrium dialysis. The results demonstrate that during aging of the normal bovine lens, there is a decrease in the ability of alpha crystallins to bind to gamma crystallins, consistent with the involvement of this interaction in the transparent properties of the lens. PMID- 16712839 TI - Thioredoxin induced antioxidant gene expressions in human lens epithelial cells. AB - Thioredoxin (Trx) is one of the major redox-regulating proteins. It catalyzes dithiol/disulfide exchange reactions and displays many unique intracellular and extracellular activities thereby controlling multiple mammalian cell functions. In the present study we examine the effect of exogenous Trx on the expression of several antioxidant genes in human lens epithelial (HLE B3) cells. mRNA levels for gene expression were monitored by RT-PCR and real-time PCR while protein levels were measured by western blot analysis. We have found that recombinant human Trx (hTrx)-treated HLE B3 cells have a simultaneous increase in mRNA expressions of mitochondrial manganese superoxide dismutase (MnSOD), thioltranferase 1 (TTase 1) or glutaredoxin 1 (Grx1), mitochondrial thioltransferase (TTase 2) or glutaredoxin 2 (Grx2), and thioredoxin peroxidase IV (Prx IV). The increased MnSOD and TTase 1 mRNA expressions were accompanied with their respective increases in protein levels. Other antioxidant genes, including Cu/ZnSOD, catalase, glutathione peroxidase 1 (GPx1), thioredoxin reductase 1 (TrxR1), thioredoxin peroxidase III (Prx III), and gamma-glutamyl cysteine synthetase were not affected. The ability of Trx to induce selectively these antioxidant genes in the absence of oxidative stress suggest a cytokine/growth factor-like new physiological role of hTrx in HLE B3 cells. Our data also provide evidence of a strong antioxidant defense system in HLE B3 cells that can be activated by extracellular hTrx, as well as of a possible link between the thioredoxin (Trx) and glutathione (GSH) redox regulating systems in these cells. PMID- 16712840 TI - Increased number of new neurons in the olfactory bulb and hippocampus of adult non-human primates after focal ischemia. AB - Adult neurogenesis is modulated by growth factors, physical conditions, and other alterations in the physical microenvironment. We studied the effects of focal ischemia on neurogenesis in the subventricular zone (SVZ), olfactory bulb (OB), and hippocampal dentate gyrus (DG) (known to be persistent neurogenic regions) in the adult non-human primate, the cynomolgus monkey. Three monkeys underwent middle cerebral artery occlusion-induced focal ischemia and were given multiple BrdU injections during the first 2 weeks after ischemia. Twenty-eight days later, the animals were perfused. The number of new neurons (3182 +/- 408/mm3) in the ipsilateral DG of ischemic monkeys was 4.7-fold that in the DG of non-operated monkeys. The number of new neurons (9176 +/- 2295/mm3) in the ipsilateral olfactory bulb of ischemic monkeys was 18.0-fold that in normal olfactory bulb. These observations suggest an increase in the number of new OB neurons, as well as new DG neurons, after focal ischemia in a primate. This substantial increase in new neurons after focal ischemia could result from the enhancement of cell proliferation rather than a change in the rate of cell commitment. Of the three monkeys subjected to ischemia, only one animal possessed a unique progenitor cell type at the most anterior aspect of the ipsilateral SVZ. Within this region, a short migration (approximately 500 microm) of doublecortin-expressing immature neuronal progenitor cells was observed. PMID- 16712841 TI - The cyanobacterial principal sigma factor region 1.1 is involved in DNA-binding in the free form and in transcription activity as holoenzyme. AB - Cyanobacterial principal sigma factor, sigma(A), includes a specifically conserved cluster of basic amino acids in the amino-terminal extension called region 1.1. We found that the sigma(A) in a thermophilic cyanobacterium Thermosynechococcus elongatus BP-1 binds DNA in the absence of the core RNA polymerase and that sigma(A) lacking region 1.1 is not able to bind DNA. This indicates that, in the cyanobacterium, region 1.1 participates in DNA-binding, rather than inhibiting the interaction between free sigma and DNA, as found in other principal sigma factors of eubacteria. The results of in vitro transcription assays with the reconstituted RNA polymerase showed that region 1.1 reduces transcription activity from the cpc promoter. PMID- 16712842 TI - Evidence for distinct functions for human DNA repair factors hHR23A and hHR23B. AB - Rad23 proteins bind ubiquitinated substrates and the proteasome, consistent with an important role in protein degradation. Although human Rad23 proteins (hHR23A and hHR23B) have redundant roles in DNA repair, we determined they formed distinct interactions with proteasomes and multiubiquitinated proteins, but similar binding to Ataxin-3. Threonine-79 contributed to the weak proteasome binding property of hHR23A, and its conversion to proline (T79P), which is the residue present in hHR23B, increased proteasome interaction. We also determined that hHR23A and hHR23B could be co-purified with unique proteolytic and stress responsive factors from human breast cancer tissues, indicating that they have unique functions in vivo. PMID- 16712843 TI - Electron transfer among the CuA-, heme b- and a3-centers of Thermus thermophilus cytochrome ba3. AB - The 1-methyl-nicotinamide radical (MNA(*)), produced by pulse radiolysis has previously been shown to reduce the Cu(A)-site of cytochromes aa(3), a process followed by intramolecular electron transfer (ET) to the heme a but not to the heme a(3) [Farver, O., Grell, E., Ludwig, B., Michel, H. and Pecht, I. (2006) Rates and equilibrium of CuA to heme a electron transfer in Paracoccus denitrificans cytochrome c oxidase. Biophys. J. 90, 2131-2137]. Investigating this process in the cytochrome ba(3) of Thermus thermophilus (Tt), we now show that MNA(*) also reduces Cu(A) with a subsequent ET to the heme b and then to heme a(3), with first-order rate constants 11200 s(-1), and 770 s(-1), respectively. The results provide clear evidence for ET among the three spectroscopically distinguishable centers and indicate that the binuclear a(3) Cu(B) center can be reduced in molecules containing a single reduction equivalent. PMID- 16712845 TI - Reexamination of the cysteine residues in glucocerebrosidase. AB - Glucocerebrosidase, the deficient enzyme in Gaucher disease, catalyzes the cleavage of the beta-glycosidic linkage of glucosylceramide. A previous study on the enzyme identified three disulfide bridges and a single sulfhydryl [Lee, Y., Kinoshita, H., Radke, G., Weiler, S., Barranger, J.A. and Tomich, J.M. (1995) Position of the sulfhydryl group and the disulfide bonds of human glucocerebrosidase. J. Protein Chem. 14(3), 127-137] but recent publication of the X-ray structure identifies only two disulfide bridges with three free sulfhydryls [Dvir, H., Harel, M., McCarthy, A.A., Toker, L., Silman, I., Futerman, A.H. and Sussman, J.L. (2003) X-ray structure of human acid-beta glucosidase, the defective enzyme in Gaucher disease. EMBO. 4(7), 704-709]. Using chemical modifications, acid cleavage and enzymatic digestion methods, we report that three free sulfhydryls exist and that the remaining four cysteines form two disulfide bonds located within the first 25 amino-terminal residues, supporting the X-ray structure. PMID- 16712844 TI - CYP4A11 is repressed by retinoic acid in human liver cells. AB - CYP4A11, the major fatty acid omega-hydroxylase in human liver is involved in the balance of lipids, but its role and regulation are both poorly understood. We studied the effects of retinoids on the regulation of CYP4A11 in the human hepatoma cell line HepaRG. Treatment of HepaRG cells with all-trans-retinoic acid resulted in a strong decrease in CYP4A11 gene expression and apoprotein content and, furthermore, was associated with a 50% decrease in the microsomal lauric acid hydroxylation activity. Such a strong suppression of CYP4A11 expression by retinoids could have a major impact on fatty acid metabolism in the liver. PMID- 16712846 TI - Human cloning. PMID- 16712847 TI - Pelvic adhesions and laparoscopic-assisted vaginal hysterectomy. PMID- 16712848 TI - Antinuclear antibodies and endometriosis. PMID- 16712849 TI - A cross-sectional survey of the symptom experience of kidney transplant recipients. AB - BACKGROUND: Previous studies have shown that being able to assess and improve kidney transplant recipients' experience of the common side effects of long-term post-operative immunosuppressive drug therapy can positively affect their treatment compliance, and thus, their quality of life. However, more needs to be learned about the factors affecting the symptom experience of kidney transplant patients in Taiwan. OBJECTIVE: The purpose of this study was to further our understanding of kidney transplant recipients' symptom experience in Taiwan in order to improve their symptom experience, drug and other therapies, and overall quality of life. METHODS: A cross-sectional study was conducted with purposive sampling of transplant recipients from a hospital in southern Taiwan. Data were collected from August 23, 2003 to June 30, 2004. One hundred subjects participated and self-reported using a structured questionnaire. The reliability (alpha) of social support and symptom experience was 0.96 and 0.78, respectively. RESULTS: The results indicated that there were significant differences in physical symptom experience by gender and herbal medicine use, and in psychosocial symptom experience by educational level and herbal medicine use. Women, on an average, had more severe physical symptoms than men. A higher level of social support and herbal medicine use was associated with less severe physical symptoms among men and women. These three factors explained 22.5% of the variance in physical symptom experience: gender (13.0%), value support (6.1%) and herbal medicine use (3.4%). The factor with the highest explanatory value for the variance in psychosocial symptom experience was herbal medicine (7.8%), its use being associated with slightly lower levels of psychosocial symptoms. CONCLUSIONS: A better understanding of kidney transplant patients' symptom experience can improve health professionals' symptom appraisal efforts, enabling them to provide better quality care, encourage greater patient compliance, and ultimately, improve the quality of life for kidney transplant recipients. PMID- 16712850 TI - The quality of parental consent for research with children: a prospective repeated measure self-report survey. AB - BACKGROUND: Researchers have ethical and legal responsibilities to ensure that individuals give informed consent to participate in research. The few studies of parental consent for paediatric research suggest there may be inadequate competence, information, understanding, or voluntariness for valid consent to occur. OBJECTIVES: To determine parents' level of understanding of the research study requirements and satisfaction with the informed consent process. PARTICIPANTS: English literate parents of children actively involved in research studies. METHODS: A repeated measures self-report survey was conducted to measure parent understanding (actual and perceived) of the study consented for and satisfaction with the informed consent process. Relationships between parents understanding of the research and their satisfaction with the consent process were explored and changes in parent understanding or satisfaction over time were described. RESULTS: Questionnaires from 109 parents were returned, representing 25 different studies. Parents demonstrated a high level of knowledge of information essential for informed consent, such as the purpose, benefits, and participant rights. Nervousness or inability to concentrate, and reading ease of the information sheet were found to relate to parents' knowledge and their perceptions of the adequacy of the consent. Parents overall reported high satisfaction with the consent process. CONCLUSIONS: These findings support and extend previous research on parental consent for research with children. They suggest areas where further research is indicated, including: the value and use of information and consent documents given to parents, the views and concerns of parents for whom English is not their first language, and further exploration of the concerns of the few dissatisfied parents. Current practices of obtaining informed consent for research lack supporting research evidence and may not be ethically justifiable. PMID- 16712851 TI - Stroke patients' experiences of sharing rooms with dementia patients in a nursing home. AB - BACKGROUND: The mixed units that place lucid and dementia patients in same rooms have been viewed to benefit the dementia patients. However, many studies report negative attitudes of lucid residents towards mixed units, and there is a scarcity of research which explores the experiences of lucid residents while sharing rooms with the dementia patients in extended care homes. Currently many special care nursing facilities have mixed units in Korea, suggesting a need to examine their effects especially on cognitively lucid patients. OBJECTIVE: To explore lived experiences of stroke patients who were sharing rooms with patients with dementia in a nursing home. DESIGN: It was a qualitative study applying a phenomenological method to explore the experiences of stroke patients. SETTINGS: Data were collected in a specialized nursing home in Korea. The nursing home provides free medical and nursing care to persons suffering either from dementia or stroke. PARTICIPANTS: Fourteen participants without cognitive deficit and who were sharing rooms with dementia patients were recruited through a purposive sampling. METHOD: In-depth interviews RESULTS: Stroke patients sharing rooms with dementia patients were being seriously affected by intense, deeply disturbing, and persistent experiences. The experiences themselves seemed to evolve over time as each patient struggled on one's own to try to make sense out of what were happening. The stroke patients ended up having a sense of resignation and anger realizing that they had no power to change the policy or the situation. The stroke patients also were distraught about what were happening to themselves - change of their own character, continuous fear of becoming demented, and becoming to devalue life. They also became indifferent to others, and seemed to have lost motivation for activity resulting in a decreased or low activity level. Several personal and environmental factors were identified that tended to increase the level of suffering. We suggest that the whole concept of mixed units require further exploration, and there is a need for a comprehensive staff education program to help the staff become aware of the problems and provide support to patients. Orientation of stroke patients regarding dementia is also suggested. PMID- 16712852 TI - Identifying outcomes from the nursing outcomes classification as indicators of quality of care in Korea: a modified delphi study. AB - BACKGROUND: The evaluation of patient outcomes as a measure of quality control of patient care is being adopted in Korean hospitals. Since nursing care contributes to the bulk of patient care, it is important to identify nursing-sensitive patient outcomes, hereafter referred to as 'nursing outcomes', that will be useful in the evaluation of nursing care. OBJECTIVE: This study was conducted to identify nursing outcomes included within the Nursing Outcomes Classification (NOC) that are most sensitive for the evaluation of nursing care in Korean hospitals as well as being observable and measurable. DESIGN: Delphi technique modified for this study was used to gain a consensus from Korean nursing experts. SETTINGS: Participants were recruited from general hospitals in Korea. PARTICIPANTS: Two hundred and thirty nurses working for Quality Improvement (QI) and Continuous Quality Improvement (CQI) programmes were chosen as the nurse experts for this study. METHODS: Three rounds of data collection from all participants was undertaken. In the first data collection, the sensitivity of 260 NOC nursing outcomes (Johnson, et al., 2000. Iowa Outcomes Projects: Nursing Outcomes Classification (NOC). C.V. Mosby, St. Louis) was examined, and more highly nursing sensitive ones were selected. In the second and third data collection phases, nursing outcomes which are most useful for the evaluation of nursing care were selected. RESULTS: Vital Signs Status, Knowledge: Infection Control, Pain Control, Safety Behavior: Fall Prevention, and Infection Status were identified as the five most useful nursing outcomes for the evaluation of nursing care in hospitals. CONCLUSIONS: The nursing outcomes identified highly useful for the evaluation of nursing care in this study can be used effectively for the quality management of nursing care in Korea. PMID- 16712853 TI - Plasma leptin: associations with metabolic, inflammatory and haemostatic risk factors for cardiovascular disease. AB - BACKGROUND AND AIM: Leptin, an adipocyte-derived protein, regulating food intake and metabolism has been implicated in the development of coronary heart disease. We have examined the relationship between leptin and vascular risk factors including insulin resistance, metabolic, inflammatory and haemostatic risk factors. METHODS AND RESULTS: The study was carried out in 3640 non-diabetic men aged 60-79 years drawn from general practices in 24 British towns and who were not on warfarin. Leptin was strongly positively correlated with waist circumference (r=0.58; p<0.0001). Leptin concentrations decreased significantly with increasing physical activity and were lowered in cigarette smokers and elevated in men with pre-existing coronary heart disease and stroke; alcohol intake showed no association with leptin concentration. After adjustment for waist circumference and these lifestyle factors, increased leptin was independently associated with significant increases in insulin resistance, triglycerides, inflammatory markers (interleukin-6, C-reactive protein, fibrinogen, plasma viscosity), coagulation factor VIII, endothelial markers von Willebrand factor, tissue plasminogen activator, and fibrin D-dimer levels; and a decrease in HDL-cholesterol. No association was seen between leptin and blood pressure, total cholesterol, glucose or white cell count after adjusting for waist circumference. Further adjustment for insulin resistance abolished the relationships between leptin and triglycerides and HDL-cholesterol, weakened the associations with the haemostatic factors although they remained significant, but made minor differences to the associations with inflammatory markers. CONCLUSION: Plasma leptin is associated with insulin resistance, inflammation and disturbances in haemostasis independent of waist circumference, suggesting possible pathways by which leptin may influence risk of cardiovascular disease. PMID- 16712854 TI - Racial difference in endothelial function: role of the infective burden. AB - There is much evidence to suggest the existence of racial differences between blacks and whites in the behaviour of endothelial function. Infective state, sustained by viral or bacterial agents, may injure the endothelial surface favouring the onset and progression of atherosclerotic process, mainly by an inflammatory mechanism. The aim of the study was to investigate endothelial function, expressed as brachial flow-mediated vasodilation (FMV), in black and white healthy subjects, along with antibody titer to cytomegalovirus, hepatitis virus (B, C), herpes virus-1 and 2, Epstein-Barr, Chlamydia pneumoniae and the expression of adhesion molecules. We enrolled 22 young (mean age 27+/-8 years) healthy subjects of black race (10 males) and 20 healthy young subjects (10 males, mean age 28+/-9 years) of white race. Total infectious burden (TIB) was defined as the number of serological positive infections. Black subjects have a reduced brachial FMV (6.9+/-3.5% versus 11.6+/-3.0%, p<0.01) and increased values of hsCRP (0.35+/-0.15 mg/dL versus 0.07+/-0.08 mg/dL, p<0.05), white cells (8578+/-1041/mmc versus 5833+/-998/mmc, p<0.01) and adhesion molecules (respectively: sVCAM-1 945+/-142 versus 779+/-93, sICAM-1 534+/-107 ng/mL versus 325+/-80 ng/mL; both p<0.01) in comparison to white subjects. The total infectious burden in black race was significantly higher than in white race (5+/ 1 versus 2+/-1, p<0.01). At the univariate analysis, brachial FMV was significantly related to the levels of adhesion molecules (respectively: sVCAM-1 r=-0.49; sICAM-1 r=-0.50, both p<0.05), hsCRP (r=-0.47, p<0.05) and white blood cells (r=-0.43, p<0.05). TIB was associated with brachial FMV (r=-0.64, p<0.05), sVCAM-1 (r=0.55, p<0.05) and hsCRP (r=0.47, p<0.05). At the multivariate analysis the only predictive variables for brachial FMV were hsCRP, TIB and brachial diameter (respectively: beta=-0.49, -0.19, -0.54, all p<0.05). This study confirms that endothelial reactivity is impaired in young African black patients; moreover its behavior is strictly related to the inflammatory state and to the total infectious burden. PMID- 16712855 TI - Endothelial function and arterial stiffness in sickle-thalassemia patients. AB - BACKGROUND: Homozygous sickle-cell anemia and beta-thalassemia are characterized by impaired endothelial function, while data on arterial stiffness have hitherto been conflicting. We sought to investigate aortic elastic properties and endothelial function in sickle-thalassemia, which combines molecular and clinical features of the above conditions. METHODS AND RESULTS: Forty-seven sickle thalassemia patients, younger than 45 years, with preserved left ventricular (LV) function and no history of smoking, systemic or pulmonary hypertension, diabetes mellitus, dyslipidemia or thyroid disease, along with 40 healthy controls were studied. Aortic strain, distensibility and stiffness index were calculated by echocardiographically-obtained aortic root diameters. Brachial artery endothelial function was assessed by ultrasonographic evaluation of flow-mediated dilatation (FMD) and nitrate-mediated dilatation (NMD). Left ventricle was assessed by echocardiography. Patients had an impaired FMD (4.2+/-2.9% versus 9.2+/-3.8% in controls, p<0.001) with a preserved NMD (16.9+/-5.6% versus 15.2+/-4.8% in controls, p>0.05). Aortic strain and distensibility were lower and aortic stiffness index was higher in patients compared to controls (8.1+/-4.6 versus 5.8+/-2.9, p<0.01). Indexed LV diameters and mass were higher in patients. Systolic LV function was preserved, while 14.9% of patients had an impaired relaxation transmitral inflow pattern. Patients' LV mass index and diastolic mitral E wave deceleration time were positively correlated with aortic stiffness index (p<0.001). CONCLUSION: Sickle-thalassemia is characterized by a complex vasculopathy, consisting of endothelial dysfunction and increased arterial stiffness, with a global effect on cardiovascular function. PMID- 16712856 TI - Stepwise reduction of functional spinal structures increase vertebral translation and intradiscal pressure. AB - To date, there are only a few studies that provide data to efficiently calibrate finite element models for the spine due to its complexity. In a recent study, we quantified the range of motion rotation and the lordosis angle. This paper provides complementary results regarding two more parameters, intradiscal pressure and vertebral translation. All parameters were obtained as a function of stepwise anatomical reduction, loading direction and magnitude. Eight lumbar spinal segments (L4-5) with a median age of 52 years (38-59 years) and no signs of disc degeneration were used for the in vitro testing. A miniaturized pressure probe was implanted into the nucleus. An ultrasound-based motion-tracking system was employed to record spatial movements of several landmarks on the specimens. The center of L4, the anterior, posterior, left and right point of the lower endplate of L4 were digitized as landmarks and its translation was determined. Specimens were loaded with pure moments (1-10Nm) in the three principal anatomical planes at a loading rate of 1.0 degrees /s. Anatomy was stepwise reduced by cutting different ligaments, facet capsules and joints and removing nucleus. Translation analysis showed that the L4 center point had its largest displacement in sagittal direction and almost none vertically. Removal of the supra- and interspinous, flaval ligaments showed a slight increase and further removal of structures, a higher increase of translation. Axial rotation also was accompanied with L4 to elevate when torsion was applied. This effect was found to be larger with progressing defects. Nucleotomy exhibited the most unstable situation for specimens. Results of the intradiscal pressure indicated a large increase after removing the facet capsules and joints. Furthermore, it was found that intradiscal pressure correlated well with data of range of motion for rotation. Predicting and simulating clinical defects, surgical intervention or treatment methods requires a well performed calibration based on in vitro data, whereas it is important to adapt all including structures with as many known parameters as possible. Results provided by these studies may be used as a database for researchers aiming to calibrate or validate finite element models of L4-5 segments. PMID- 16712857 TI - Quantitation of the nearest-neighbour effects of amino acid side-chains that restrict conformational freedom of the polypeptide chain using reversed-phase liquid chromatography of synthetic model peptides with L- and D-amino acid substitutions. AB - Side-chain backbone interactions (or "effects") between nearest neighbours may severely restrict the conformations accessible to a polypeptide chain and thus represent the first step in protein folding. We have quantified nearest-neighbour effects (i to i+1) in peptides through reversed-phase liquid chromatography (RP HPLC) of model synthetic peptides, where L- and D-amino acids were substituted at the N-terminal end of the peptide sequence, adjacent to a L-Leu residue. These nearest-neighbour effects (expressed as the difference in retention times of L- and D-peptide diastereomers at pHs 2 and 7) were frequently dramatic, depending on the type of side-chain adjacent to the L-Leu residue, albeit such effects were independent of mobile phase conditions. No nearest-neighbour effects were observed when residue i is adjacent to a Gly residue. Calculation of minimum energy conformations of selected peptides supported the view that, whether a L- or D-amino acid is substituted adjacent to L-Leu, its orientation relative to this bulky Leu side-chain represents the most energetically favourable configuration. We believe that such energetically favourable, and different, configurations of L- and D-peptide diastereomers affect their respective interactions with a hydrophobic stationary phase, which are thus quantified by different RP-HPLC retention times. Side-chain hydrophilicity/hydrophobicity coefficients were generated in the presence of these nearest-neighbour effects and, despite the relative difference in such coefficients generated from peptides substituted with L- or D-amino acids, the relative difference in hydrophilicity/hydrophobicity between different amino acids in the L- or D-series is maintained. Overall, our results demonstrate that such nearest-neighbour effects can clearly restrict conformational space of an amino acid side-chain in a polypeptide chain. PMID- 16712858 TI - Use of near infrared emission spectroscopy in the study of supporting materials and stationary phases for liquid chromatography. AB - Near infrared emission spectroscopy (NIRES) has been investigated in the study of different materials employed in liquid chromatography. The samples were heated in a nitrogen atmosphere and the emission spectra were obtained using a lab-made NIRES instrument. Through principal component analysis (PCA) using the raw emission spectra, it was possible to distinguish different materials according to their physical and/or chemical characteristics. Linear relationships between emissivity spectra and the contents of the coating material or the specific surface areas was observed for stationary phases or bare silicas, respectively. Furthermore, the thermal stability of stationary phases could be followed in real time. PMID- 16712859 TI - Nucleotide-mimetic synthetic ligands for DNA-recognizing enzymes One-step purification of Pfu DNA polymerase. AB - The commercial availability of DNA polymerases has revolutionized molecular biotechnology and certain sectors of the bio-industry. Therefore, the development of affinity adsorbents for purification of DNA polymerases is of academic interest and practical importance. In the present study we describe the design, synthesis and evaluation of a combinatorial library of novel affinity ligands for the purification of DNA polymerases (Pols). Pyrococcus furiosus DNA polymerase (Pfu Pol) was employed as a proof-of-principle example. Affinity ligand design was based on mimicking the natural interactions between deoxynucleoside triphosphates (dNTPs) and the B-motif, a conserved structural moiety found in Pol I and Pol-II family of enzymes. Solid-phase 'structure-guided' combinatorial chemistry was used to construct a library of 26 variants of the B-motif-binding 'lead' ligand X-Trz-Y (X is a purine derivative and Y is an aliphatic/aromatic sulphonate or phosphonate derivative) using 1,3,5-triazine (Trz) as the scaffold for assembly. The 'lead' ligand showed complementarity against a Lys and a Tyr residue of the polymerase B-motif. The ligand library was screened for its ability to bind and purify Pfu Pol from Escherichia coli extract. One immobilized ligand (oABSAd), bearing 9-aminoethyladenine (AEAd) and sulfanilic acid (oABS) linked on the triazine scaffold, displayed the highest purifying ability and binding capacity (0,55 mg Pfu Pol/g wet gel). Adsorption equilibrium studies with this affinity ligand and Pfu Pol determined a dissociation constant (K(D)) of 83 nM for the respective complex. The oABSAd affinity adsorbent was exploited in the development of a facile Pfu Pol purification protocol, affording homogeneous enzyme (>99% purity) in a single chromatography step. Quality control tests showed that Pfu Pol purified on the B-motif-complementing ligand is free of nucleic acids and contaminating nuclease activities, therefore, suitable for experimental use. PMID- 16712860 TI - Thermal stability of glucose and other sugar aldoses in normal phase high performance liquid chromatography. AB - Analysis of glucose and other simple sugars are often performed by use of normal phase HPLC methods with acetonitrile as major eluent. The present results clearly show that column temperature plays an important role with respect to chromatographic performance and detection limits of glucose when using a specific carbohydrate column. A change in column temperature from 25 to 45 degrees C reduced the detection of glucose (with ELSD) by more than 41%, whereas the detection of other sugar aldoses (galactose, xylose and rhamnose) were suppressed even more. By increase of column temperature to 70 degrees C the detector signal of glucose was found to be less than 2% compared to that obtained at 20 degrees C. Neither fructose nor sucrose showed similar correlation between column temperature and detection. The rate of decreased response is not dependent on sample concentration or the ELSD settings. The results express the importance of accurate temperature control in the analysis of sugar aldoses, and also values low column temperatures for samples with low concentrations of sugar aldoses in order to improve detection. PMID- 16712861 TI - Application of a column selection system and DryLab software for high-performance liquid chromatography method development. AB - This paper describes a strategy for the development of chromatographic methods for drug candidates based upon the use of simple MS compatible mobile phases and optimization of the chromatographic selectivity through variations of the stationary phase and mobile phase pH. The strategy employs an automated column selection system and a series of HPLC columns, varying in hydrophobicity and silanol activity, in combination with DryLab software to develop chromatographic methods for the separation of mixtures of bupivacaine and its metabolites; acidic, basic, and neutral compounds; and atenolol, nitrendipine, and their degradation products. PMID- 16712862 TI - Ten cases of feline mesothelioma: an immunohistochemical and ultrastructural study. AB - In the cat only 10 cases of mesothelioma, mainly of the peritoneum, have been previously reported. This paper describes a further 10 cases, eight pleural and two peritoneal, in males and females aged 1-17 years. Histologically, five tumours were epithelial, three fibrosarcomatous and two biphasic. Immunohistochemical markers used in human pathology for the identification of mesotheliomas include vimentin, cytokeratin (CK) AE1/AE3, HBME-1, CK 5/6, calretinin, thrombomodulin, carcinoembryonic antigen (CEA), CD15, E-cadherin and desmin. All 10 feline mesotheliomas were positive for vimentin and CK AE1/AE3, six were positive for HBME-1, two for CK5/6, three for CEA and four for E cadherin. All were negative for desmin and calretinin. Antibodies to thrombomodulin and CD15 failed to cross-react with feline tissues. Electron microscopy, performed in four cases, revealed microvillar structures, desmosomes and intracytoplasmic lumina, confirming its value as a diagnostic tool. The study showed that mesothelial marker antibodies commonly used in human patients can be used for the diagnosis of feline mesothelioma, preferably as a panel of antibodies rather than only one. PMID- 16712863 TI - Pathogenesis of bovine neosporosis. AB - The protozoan parasite Neospora caninum is a major pathogen of cattle and dogs, being a significant cause of abortion in cattle in many countries. It is one of the most efficiently transmitted parasites, with up to 90% of cattle infected in some herds. The pathogenesis of abortion due to Neospora is complex and only partially understood. Losses occur after a primary infection during pregnancy but more commonly as the result of recrudescence of a persistent infection during pregnancy. Parasitaemia is followed by invasion of the placenta and fetus. It is suggested that abortion occurs when primary parasite-induced placental damage jeopardises fetal survival directly or causes release of maternal prostaglandins that in turn cause luteolysis and abortion. Fetal damage may also occur due to primary tissue damage caused by the multiplication of N. caninum in the fetus or due to insufficient oxygen/nutrition, secondary to placental damage. In addition, maternal immune expulsion of the fetus may occur associated with maternal placental inflammation and the release of maternal pro-inflammatory cytokines in the placenta. Thus N. caninum is a primary pathogen capable of causing abortion either through maternal placental inflammation, maternal and fetal placental necrosis, fetal damage, or a combination of all three. The question of how N. caninum kills the fetus exposes the complex and finely balanced biological processes that have evolved to permit bovine and other mammalian pregnancies to occur. Defining these immunological mechanisms will shed light on potential methods of control of bovine neosporosis and enrich our understanding of the continuity of mammalian and protozoal survival. PMID- 16712864 TI - Relative importance of CD4+ and CD8+ T cells in the resolution of Chlamydophila abortus primary infection in mice. AB - The role of the specific cellular immune response is well established in Chlamydiaceae infections, but the importance of each T-cell subset seems to be species-dependent. This study was designed to clarify the role of T-cell subsets in the response to Chlamydophila abortus primary infection. C57BL/6 mice were depleted of CD4+ or CD8+, or both, by monoclonal antibody injections and subsequently infected with C. abortus. Mice were killed at intervals and samples were collected for bacteriological and histopathological analysis. Also carried out were spleen cell culture, cytokine quantification, immunolabelling for C. abortus antigen, and a TUNEL assay for apoptosis. CD8+ T cell-depleted mice all died within 12 days of C. abortus infection, while no mortality was observed in the other groups; surprisingly, CD4+ T cell-depleted mice showed lower morbidity (expressed as weight loss) than did a non-depleted (control) group. CD8+ T cell depleted mice also differed from the other groups in showing a significantly higher chlamydial burden in the liver. CD8+ T cell-depleted mice also had a higher number of apoptotic cells in hepatic inflammatory foci and showed exacerbated IFN-gamma production by spleen cells after specific stimulation. Simultaneous depletion of both T-cell subpopulations led to a chronic infection, but not to early mortality. It is concluded that CD8+ T cells may play a role in the regulatory control of the CD4+ T-cell response and may have a direct cytotoxic or IFN-gamma-mediated effect on infected cells. PMID- 16712865 TI - Lung lesions in SCID-bo and SCID-bg mice after intratracheal inoculation with wild-type or leucotoxin-deficient mutant strains of Mannheimia haemolytica serotype 1. AB - The purpose of this study was to investigate SCID-bg mice engrafted with bovine haematolymphoid tissues (SCID-bo) as a model for studying bovine Mannheimia haemolytica serotype 1- induced pneumonia, in which leucotoxin (LKT) plays a major role. In experiment A, SCID-bo and SCID-bg mice were inoculated intratracheally with either (1) phosphate-buffered saline (PBS), (2) M. haemolytica wild-type strain 89010807N ("LKT(+)WT"), (3) a M. haemolytica leucotoxin-deficient mutant of strain 89010807N ("LKT(-)mutant"), or (4) the M. haemolytica wild-type Oklahoma strain. Mice were killed for examination at intervals between 20 and 44h after inoculation. Lung lesions consisted of thickened alveolar septa and neutrophil and macrophage infiltrates in the bronchioles and alveoli. Lung lesion scores in the SCID-bo mice inoculated with LKT(+)WT or LKT(-) mutant were significantly (P<0.05) greater than those of the PBS control group, but the two bacterial strains produced results that did not differ significantly. M. haemolytica was isolated from lung, liver and spleen after inoculation but less frequently as time progressed. In experiment B, SCID bg mice were inoculated intratracheally with live LKT(+)WT or formalin-killed LKT(+)WT and killed 24, 48 or 96 h later. Lung lesions were histologically similar to those observed in experiment A; however, there were no significant differences in the lung lesion scores between groups. It was concluded that the lesions seen in this study were probably not due to LKT, and that the SCID-bo mouse does not provide a good rodent model for bovine pneumonia. PMID- 16712866 TI - Statistical determination of threshold for cellular division in the CFSE-labeling assay. AB - The combination of flow cytometry and carboxyl fluorescent succinimidyl ester (CFSE) labeling techniques has been widely used in the study of cellular proliferation, including measurement of the percentage of proliferated cells and the number of cell divisions undergone by proliferated cells. However, the smallest numbers that represent true cell division rather than experimental variation are not known. To define a threshold that separates true proliferation from experimental variation, we performed a large number of replicate CFSE labeling experiments using polyclonal stimulation, obtained the percentages of proliferated cells using ModFit software, and then analyzed these data using several statistical methods. Our results indicate that the threshold of proliferation lies between 0.071% (95% confidence) and 0.114% (99% confidence) of total CFSE-labeled cells under our laboratory conditions. We offer our methods presented here for other investigators to calculate a threshold in their own CFSE labeling experiments. PMID- 16712867 TI - Virus-like particles in venom of Meteorus pulchricornis induce host hemocyte apoptosis. AB - Ultrastructural studies on the reproductive tract and venom apparatus of a female braconid, Meteorus pulchricornis, revealed that the parasitoid lacks the calyx region in its oviduct, but possesses a venom gland with two venom gland filaments and a venom reservoir filled with white and cloudy fluid. Its venom gland cell is concaved and has a lumen filled with numerous granules. Transmisson electron microscopic (TEM) observation revealed that virus-like particles (VLPs) were produced in venom gland cells. The virus-like particle observed in M. pulchricornis (MpVLP) is composed of membranous envelopes with two different parts: a high-density core and a whitish low-density part. The VLPs of M. pulchricornis is also found assembling ultimately in the lumen of venom gland cell. Microvilli were found thrusting into the lumen of the venom gland cell and seem to aid in driving the matured MpVLPs to the common duct of the venom gland filament. Injection of MpVLPs into non-parasitized Pseudaletia separata hosts induced apoptosis in hemocytes, particularly granulocytes (GRs). Rate of apoptosis induced in GRs peaked 48h after VLP injection. While a large part of the GR population collapsed due to apoptosis caused by MpVLPs, the plasmatocyte population was minimally affected. The capacity of MpVLPs to cause apoptosis in host's hemocytes was further demonstrated by a decrease ( approximately 10-fold) in ability of host hemocytes to encapsulate fluorescent latex beads when MpVLPs were present. Apparently, the reduced encapsulation ability was due to a decrease in the GR population resulting from MpVLP-induced apoptosis. PMID- 16712869 TI - Domain organization and crystal structure of the catalytic domain of E.coli RluF, a pseudouridine synthase that acts on 23S rRNA. AB - Pseudouridine synthases catalyze the isomerization of uridine to pseudouridine (Psi) in rRNA and tRNA. The pseudouridine synthase RluF from Escherichia coli (E.C. 4.2.1.70) modifies U2604 in 23S rRNA, and belongs to a large family of pseudouridine synthases present in all kingdoms of life. Here we report the domain architecture and crystal structure of the catalytic domain of E.coli RluF at 2.6A resolution. Limited proteolysis, mass spectrometry and N-terminal sequencing indicate that RluF has a distinct domain architecture, with the catalytic domain flanked at the N and C termini by additional domains connected to it by flexible linkers. The structure of the catalytic domain of RluF is similar to those of RsuA and TruB. RluF is a member of the RsuA sequence family of Psi-synthases, along with RluB and RluE. Structural comparison of RluF with its closest structural homologues, RsuA and TruB, suggests possible functional roles for the N-terminal and C-terminal domains of RluF. PMID- 16712868 TI - Characterization of digestive proteolytic activity in Lygus hesperus Knight (Hemiptera: Miridae). AB - The tarnished plant bug, Lygus hesperus Knight, is a pest that causes considerable economic losses to vegetables, cotton, canola, and alfalfa. Detailed knowledge of its digestive physiology will provide new opportunities for a sustainable pest management approach to control this insect. Little is known about the different protease class contributions to the overall digestion of a specific protein. To this end, the proteolytic activities in female adult L. hesperus salivary gland and midgut homogenates were quantified over a range of pH's and time points, and the contribution of different classes of proteases to the degradation of FITC-casein was determined. In the salivary gland, serine proteases were the predominant class responsible for caseinolytic activity, with the rate of activity increasing with increasing pH. In contrast, both aspartic and serine proteases contributed to caseinolytic activity in the midgut. Aspartic protease activity predominated at pH 5.0 and occurred immediately after incubation, whereas serine protease activity predominated at pH 7.5 after a 9h delay and was resistant to aprotinin. The salivary serine proteases were distinctly different from midgut serine proteases, based on the tissue-specific differential susceptibility to aprotinin and differing pH optima. Collectively, the caseinolytic activities complement one another, expanding the location and pH range over which digestion can occur. PMID- 16712870 TI - First report of two Taiwanese siblings with sialidosis type I: a 10-year follow up study. AB - We report the clinical features, electrophysiological findings and genetic characteristics of the first two Taiwanese siblings ever reported with sialidosis type I. We also provide a 10-year follow-up result. Enzymological analysis revealed a primary sialidase deficit. The back-averaged electroencephalography demonstrated myoclonic jerk-related cortical activities and the somatosensory evoked potential studies revealed giant cortical components. During the 10-year follow-up, the brain magnetic resonance images of the younger brother remained normal, whereas they showed mild cerebellar atrophy in the older sister. Macular cherry red spots were absent in both siblings. However, visual evoked potential revealed progressively prolonged latencies of P100 bilaterally, which was consistent with progressive deterioration of the siblings' visions. DNA analysis showed that the siblings had a homozygous missense point mutation c.544A-->G (Ser182Gly) in the exon 3 of the alpha-N-acetyl-neuraminidase (NEU1) gene. The mutation is predicted to cause a decreased sialidase activity but the mutant sialidase can still be targeted to the lysosomes, which may correlate with the mild clinical phenotypes and absent cherry red spots in the siblings. PMID- 16712871 TI - Reconciling sexual selection to species recognition: a process-based model of mating decision. AB - Mating signals often encode information important for both species recognition and mate quality assessment and endure selection pressures that combine both stabilizing and directional components. Here, we present a family of models of mate preference for multiple-message signals. Our models are process based rather than purely normative, they assume the existence of one (or more) "utility function" that order signals along a scale of perceived appropriateness, and interpret preferences either as the differential probability of signals recognition or as the combined effect of differential recognition and direct comparison between signal alternatives. These models show the critical role played by the proximate mechanisms of information processing in influencing the ultimate function of female mate choice. They show that if preferences are an emergent property of the way animals recognize signals then species recognition and mate quality assessment are expected to constrain each other severely and to limit the overall discrimination power of the system. In contrast, if preferences result from two computational processes, recognition and comparison, the constraining effects of species recognition and mate quality assessment are sensibly reduced. In these cases, females may improve discrimination in mate quality by adopting permissive recognition rules and limiting the risks of heterospecific mating. PMID- 16712872 TI - Modeling the glucose-insulin regulatory system and ultradian insulin secretory oscillations with two explicit time delays. AB - In the glucose-insulin regulatory system, ultradian insulin secretory oscillations are observed to have a period of 50-150 min. After pioneering work traced back to the 1960s, several mathematical models have been proposed during the last decade to model these ultradian oscillations as well as the metabolic system producing them. These currently existing models still lack some of the key physiological aspects of the glucose-insulin system. Applying the mass conservation law, we introduce two explicit time delays and propose a more robust alternative model for better understanding the glucose-insulin endocrine metabolic regulatory system and the ultradian insulin secretory oscillations for the cases of continuous enteral nutrition and constant glucose infusion. We compare the simulation profiles obtained from this two time delay model with those from the other existing models. As a result, we notice many unique features of this two delay model. Based on our intensive simulations, we suspect that one of the possibly many causes of ultradian insulin secretion oscillations is the time delay of the insulin secretion stimulated by the elevated glucose concentration. PMID- 16712873 TI - Pre-clinical toxicology and pathology of 9-(2'-hydroxyethylamino)-4-methyl-1 nitroacridine (C-1748), a novel anti-cancer agent in male Beagle dogs. AB - We have developed a group of 4-substituted-1-nitroacridines with potent anti tumor activity against prostate cancer and less toxic than parent 1 nitroacridines. The most active 9-(2'-hydroxyethylamino)-4-methyl-1-nitroacridine (C-1748) was selected for pre-clinical studies. The current study was undertaken to evaluate clinical and/or morphological adverse effects of C-1748 as a single intravenous dose at concentrations ranging from 0.16 to 4.6 mg/kg administered to male Beagle dogs. The maximum tolerated dose was 1.5 mg/kg. Emesis was observed in all groups lasting an average of 30 min to 12 h post-dosing. At high dose, extreme aggression was observed in one dog followed by disorientation and depression lasting for 48 h a frequent observation with chemotherapy. Reductions in platelets and white blood cells were observed which was similar to that seen with other chemotherapeutic agents. A compensatory hyperplasia of lymph nodes and a transient and limited extravasation in the intestinal mucosa were also observed. Increases in aspartate aminotransferase, alkaline phosphatase and creatine phosphokinase were transient with normal levels restored by day 9. These enzyme increases were accompanied by epithelial hypertrophy of larger bile ductules in the periportal triads of the liver. The low toxicity profile and high tumor target activity make this novel class of drug a promising chemotherapeutic agent. PMID- 16712874 TI - HMG-CoA reductase inhibitor fluvastatin prevents angiotensin II-induced cardiac hypertrophy via Rho kinase and inhibition of cyclin D1. AB - HMG-CoA reductase inhibitors, so called statins, decrease cardiac events. Previous studies have shown that HMG-CoA reductase inhibitors inhibit cardiomyocyte hypertrophy in vitro and in vivo by blocking Rho isoprenylation. We have shown that the G1 cell cycle regulatory proteins cyclin D1 and Cdk4 play important roles in cardiomyocyte hypertrophy. However, the relation between Rho and cyclin D1 in cardiomyocyte is unknown. To investigate whether HMG-CoA reductase inhibitors prevent cardiac hypertrophy through attenuation of Rho and cyclin D1, we studied the effect of fluvastatin on angiotensin II-induced cardiomyocyte hypertrophy in vitro and in vivo. Angiotensin II increased the cell surface area and [(3)H]leucine uptake of cultured neonatal rat cardiomyocytes and these changes were suppressed by fluvastatin treatment. Angiotensin II also induced activation of Rho kinase and increased cyclin D1, both of which were also significantly suppressed by fluvastatin. Specific Rho kinase inhibitor, Y-27632 inhibited angiotensin II-induced cardiomyocyte hypertrophy and increased cyclin D1. Overexpression of cyclin D1 by adenoviral gene transfer induced cardiomyocyte hypertrophy, as evidenced by increased cell size and increased protein synthesis; this hypertrophy was not diminished by concomitant treatment with fluvastatin. Infusion of angiotensin II to Wistar rats for 2 weeks induced hypertrophic changes in cardiomyocytes, and this hypertrophy was prevented by oral fluvastatin treatment. These results show that an HMG-CoA reductase inhibitor, fluvastatin, prevents angiotensin II-induced cardiomyocyte hypertrophy in part through inhibition of cyclin D1, which is linked to Rho kinase. This novel mechanism discovered for fluvastatin could be revealed how HMG-CoA reductase inhibitors are preventing cardiac hypertrophy. PMID- 16712875 TI - Differential regulation of CC chemokine receptors by 9-cis retinoic acid in the human mast cell line, HMC-1. AB - Mast cells are well known as effector cells in a variety of inflammatory diseases, including asthma as well as other allergic disorders. The precise role of 9-cis retinoic acid (9CRA) in mast cells is not understood despite the accepted fact that 9CRA regulates inflammatory responses and neutrophil differentiation. In this study, we investigated the effects of 9CRA on the expression of CC chemokine receptors in the human mast cell line, HMC-1. 9CRA selectively inhibits the CCR2 mRNA level and increases the CCR3 mRNA level in both a time and dose dependent manner. Other CC chemokine receptors, including CCR1, CCR4 and CCR5 are not altered by treatment with 9CRA. Both TNF-alpha and LPS, known pro-inflammatory molecules, have no effect on mRNA levels of CC chemokine receptors. For surface expression, 9CRA decreased the CCR2 level but had no effect on the CCR3 level. 9CRA inhibited the chemotactic activity in response to the CCR2-dependent chemokine, MCP-1/CCL2 but not in response to CCR3 specific chemokine, eotaxin/CCL11. 9CRA decreased spontaneous homotype clustering. Therefore, our results demonstrate that 9CRA differentially decreases both CCR2 expression and chemotactic ability of HMC-1 cells, and may regulate the inflammatory effects of mast cells. PMID- 16712876 TI - Gastroprotective and ulcer-healing activity of oleanolic acid derivatives: in vitro-in vivo relationships. AB - The triterpene oleanolic acid 1 and its semisynthetic derivatives 2-7 were assessed for gastroprotective and ulcer-healing effect using human epithelial gastric cells (AGS) and human lung fibroblasts (MRC-5). The ability of the compounds to protect the AGS cells against the damage induced by sodium taurocholate (NaT), to stimulate the cellular reduced glutathione (GSH) and prostaglandin E(2) content, to enhance AGS and MRC-5 cell proliferation and to scavenge superoxide anion in vitro was studied. The cytotoxicity of the compounds was assessed towards MRC-5 and AGS cells. In addition, the gastroprotective activity of the compounds was assessed in vivo using the HCl/EtOH-induced ulcer model in mice. All the assayed compounds displayed a significant reduction of AGS cells damage after incubation with NaT. None of the studied compounds was active as a superoxide anion scavenger nor stimulated the GSH content in AGS cell cultures. Compounds 1, 2, 4 and 6 were able to increase the prostaglandin content in AGS cell cultures. Concerning the proliferation assays, a significant stimulating effect was observed for compounds 3 and 7 on AGS cells and for 1 and 7 on MRC-5 fibroblasts. Regarding cytotoxicity, derivatives 2, 4, 6 and 7 were less toxic than the parent compound oleanolic acid. Our results strongly support the predictive capacity of the in vitro assessment of gastroprotective activity allowing the reduction of experimental animals. PMID- 16712877 TI - Mediterranean fin whales at risk from fatal ship strikes. AB - This paper reviews and analyzes ship collision records for the relatively isolated population of fin whales in the Mediterranean Sea from 1972 to 2001. Out of 287 carcasses, 46 individuals (16.0%) were certainly killed by boats. The minimum mean annual fatal collision rate increased from 1 to 1.7 whales/year from the 1970s to the 1990s. Fatal strike events (82.2%) were reported in or adjacent to the Pelagos Sanctuary, characterized by high levels of traffic and whale concentrations. Among 383 photo-identified whales, 9 (2.4%) had marks that were attributed to a ship impact. The reported rates are unusually high for baleen whales. The high likelihood of unreported fatal strikes combined with other anthropogenic threats suggests an urgent need for a comprehensive, basin-wide conservation strategy, including ship strike mitigation requirements, like real time monitoring of whale presence and distribution to re-locate ferry routes to areas of lower cetacean density, and reducing ship speed in high cetacean density areas. PMID- 16712878 TI - Carbon speciation and composition of natural microbial communities in polluted and pristine sediments of the Eastern Mediterranean Sea. AB - Sediment samples collected from polluted (Augusta Bay) and pristine regions of the Eastern Mediterranean Sea (South Ionian Sea, Thracian Sea) were analyzed for black carbon, aliphatic hydrocarbons and phospholipid ester-linked fatty acids (PLFA). The aim of the study was to investigate the anthropogenic and biogenic inputs into the Eastern Mediterranean Sea and to evaluate the effects of refractory organic matter (e.g. black carbon) and the level of hydrocarbon contamination on benthic microbial community composition. Black carbon, generally considered to be chemically and biologically inert, comprised a significant but highly variable fraction of the sedimentary carbon pool in the analyzed sediments with a ratio to total organic carbon ranging from 0.02 to 0.66. Principal component analysis of the chemical characteristics of the sediments (organic carbon content, black carbon, bioavailable organic carbon, chlorophyll a, phaeopigments, aliphatic hydrocarbons) revealed clustering of samples along a gradient from the most productive and contaminated region of Augusta Bay to the carbon-poor and pristine sediments of the Thracian Sea. PLFA analysis revealed that gram-negative bacteria and microeukaryotes were most abundant in Augusta Bay and in the most impacted station of the Thracian Sea. The high levels of branched and odd-chain fatty acids recorded for these stations is probably linked to the elevated amounts of hydrocarbons at these stations; e.g. microbial communities may have developed the ability to degrade either naturally occurring aliphatic hydrocarbons or hydrocarbons derived from oil contamination. PMID- 16712879 TI - Comments to "Imposex in Hexaplex (Trunculariopsis) trunculus (Gastropoda: Muricidae) from the Ria Formosa Lagoon (Algarve coast--southern Portugal)". PMID- 16712880 TI - NMDA depresses glutamatergic synaptic transmission in the striatum through the activation of adenosine A1 receptors: evidence from knockout mice. AB - N-methyl-D-aspartate (NMDA) receptors play several essential roles in the physiology and pathophysiology of the brain. Their activation results in long term changes in glutamatergic synaptic transmission in several brain areas, but excessive activation of these receptors induces neurotoxicity. Some of NMDA mediated actions are critically dependent on functional interactions with the neuromodulator adenosine. In the present study, we have examined whether pharmacological activation of NMDA receptors induces long-term changes in synaptic strength in the striatum. We found that NMDA depressed the amplitude of the field excitatory postsynaptic potential/population spike (fEPSP/PS) recorded in corticostriatal mouse brain slices in a concentration-dependent manner. Inhibition of synaptic transmission was more pronounced at room temperature (22 degrees C) than at 32 degrees C and long lasting (> 2 h) depression of the fEPSP/PS was observed only at room temperature. NMDA-induced depression of the fEPSP/PS was reduced or abolished in the presence of an A1 receptor antagonist and in A1 receptor knockout mice. In addition, exogenous application of adenosine depressed fEPSP/PS amplitude in wild-type mice, but not in A1 receptor knockout mice, in a concentration-dependent manner. Our results demonstrate that NMDA depresses synaptic transmission in a concentration- and temperature-dependent manner via release of adenosine and activation of adenosine A1 receptors. PMID- 16712881 TI - Cross talk between nitric oxide and ERK1/2 signaling pathway in the spinal cord mediates naloxone-precipitated withdrawal in morphine-dependent rats. AB - Our recent study has shown activation of spinal extracellular signal-regulated kinase-1 and -2 (ERK1/2), a member of the mitogen-activated protein kinase (MAPK) family, contributes to naloxone-precipitated withdrawal and withdrawal-induced spinal neuronal sensitization in morphine-dependent rats. However, the mechanism and significance of the spinal ERK1/2 activation during morphine dependence and withdrawal remain unknown. In this study, we reported that intrathecal (i.t.) pretreatment with either the non-selective nitric oxide synthase (NOS) inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME), neuronal NOS (nNOS) inhibitor 7 nitro indazole (7-NI), or the inducible NOS (iNOS) inhibitor aminoguanidine (AG), could reduce morphine withdrawal-induced increase of phospho-ERK1/2 (pERK1/2) expression in the rat spinal cord. On the other hand, attenuation of the spinal ERK phosphorylation by the MAPK kinase (MEK) inhibitor U0126 also could inhibit the increase of nNOS and iNOS expression in the spinal cord of morphine withdrawal rats. Inhibitory expression of pERK1/2 by i.t. NOS inhibitor L-NAME, 7 NI or AG and of nNOS and iNOS by i.t. U0126 in the spinal cord were accompanied by decreased scores of morphine withdrawal and the inhibited spinal Fos protein (a maker for neuronal excitation or activation) expression induced by morphine withdrawal. These findings suggest cross talk between nitric oxide (NO) and the ERK1/2 signaling pathway mediates morphine withdrawal and withdrawal-induced spinal neuronal sensitization in morphine-dependent rats. PMID- 16712883 TI - Olfactory influences on appetite and satiety in humans. AB - Odor stimuli play a major role in perception of food flavor. Food-related odors have also been shown to increase rated appetite, and induce salivation and release of gastric acid and insulin. However, our ability to identify an odor as food-related, and our liking for food-related odors, are both learned responses. In conditioning studies, repeated experience of odors with sweet and sour tastes result in enhanced ratings of sensory quality of the paired taste for the odor on its own. More recent studies also report increased pleasantness ratings for odors paired with sucrose for participants who like sweet tastes, and conversely decreased liking and increased bitterness for quinine-paired odors. When odors were experienced in combination with sucrose when hungry, liking was not increased if tested sated, suggesting that expression of acquired liking for odors depends on current motivational state. Other studies report sensory specific satiety is seen with food-related odors. Overall, these studies suggest that once an odor is experienced in a food-related context, that odor acquires the ability to modify both preparatory and satiety-related components of ingestion. PMID- 16712882 TI - Prism adaptation improves spatial dysgraphia following right brain damage. AB - Visuo-manual adaptation to prisms produces a long-lasting improvement of visuo spatial neglect. Improvement is also observed in tasks that do not involve visuo manual component and that can all be consider to rely on a rightward (ipsilesional) orienting bias. Here, we report positive effects of prism adaptation on spatial dysgraphia, in a neglect patient following right brain damage. A long-lasting improvement concerned the right-page preference reflecting the ipsilesional bias but also the sloping lines and the broken lines reflecting visuo-constructive disorders in handwriting. Moreover, a transient improvement was also evidenced for the graphic errors. These results reinforce the idea that the process of prism adaptation may activate brain functions related to multisensory integration and higher spatial representations and show a generalization at a functional level. Prism adaptation therefore appears as useful tool in the theoretical attempt to identify the underlying 'core' mechanisms of the neglect syndrome. PMID- 16712884 TI - Neither reduced photoperiod, nor female-related social cues, nor increased maternal thermal stress result in a paternally responsive Phodopus sungorus male. AB - Neither reduced photoperiod, nor female-related social cues, nor increased maternal thermal stress result in a paternally responsive Phodopus sungorus male. Physiol Behav [Volume # (Issue #)] 000-000, 2006. -Under laboratory conditions, P. sungorus males are not needed for pup survival and growth and are unresponsive to an experimentally displaced pup. However, males have been found with young pups late in the breeding season, and other laboratories have found evidence for biparental care. Thus, paternal behavior might be expressed in response to relevant social and environmental cues. Reduced photoperiod (from 14L:10D to 12L:12D), increased ambient temperature (from 18 to 23 degrees C) and pairing with an obligate biparental heterospecific female (to remove behavioral barriers to pup contact) were tested. Although within-animal responsiveness during a standardized pup displacement increased under reduced photoperiod, and pup retrieval occurred more often in the heterospecific pairing, paternal responsiveness scores remained negative, and typical of the species, rather than positive, and indicative of paternal responsiveness. Thus, P. sungorus males did not meet criteria for paternal responsiveness in terms of nest attendance, retrieval rates, or positive paternal responsiveness scores. Pup survival and development were not affected. In spite of their seasonal proximity to pups, there is no evidence that these environmental cues and manipulations are sufficient to result in biparental care. PMID- 16712885 TI - Enhanced oxidation of flavan-3-ols and proanthocyanidin accumulation in water stressed tea plants. AB - (-)-Epicatechin (EC) and (-)-epigallocatechin gallate (EGCG), two major tea flavan-3-ols, have received attention in food science and biomedicine because of their potent antioxidant properties. In plants, flavan-3-ols serve as proanthocyanidin (PA) building blocks, and although both monomeric flavan-3-ols and PAs show antioxidant activity in vitro, their antioxidant function in vivo remains unclear. In the present study, EC quinone (ECQ) and EGCG quinone (EGCGQ), the oxidation products of EC and EGCG, increased up to 100- and 30-fold, respectively, in tea plants exposed to 19 days of water deficit. Oxidation of EC and EGCG preceded PAs accumulation in leaves, which increased from 35 to 53 mg gDW(-1) after 26 days of water deficit. Aside from the role monomeric flavan-3 ols may play in PAs biosynthesis, formation of ECQ and EGCGQ strongly negatively correlated with the extent of lipid peroxidation in leaves, thus supporting a protective role for these compounds in drought-stressed plants. Besides demonstrating flavonoid accumulation in drought-stressed tea plants, we show for the first time that EC and EGCG are oxidized to their respective quinones in plants in vivo. PMID- 16712886 TI - Chemical and biological characterisation of sapinopyridione, a phytotoxic 3,3,6 trisubstituted-2,4-pyridione produced by Sphaeropsis sapinea, a toxigenic pathogen of native and exotic conifers, and its derivatives. AB - A phytotoxic trisubstituted 2,4-pyridione, named sapinopyridione, was isolated from the culture filtrates of Sphaeropsis sapinea, a fungal pathogen of conifers occurring world-wide. Three strains were isolated from two cypress species. Strain D-55 isolated from Cupressus sempervirens resulted high producer of sapinopyridione (12.3 mg l(-1)), whereas strain D-54 isolated from the same cypress species was low producer (1.1 mg l(-1)); strain D-50 isolated from C. macrocarpa was intermediate producer (5.4 mg l(-1)). Sapinopyridione was characterised by spectroscopic and chemical methods, as the 6-methyl-2-(2-methyl 1-oxobutyl)-1-oxa-5-azaspiro[2.5]oct-6-ene-4,8-dione. The structure was supported by the preparation of three key derivatives, whose phytotoxic and antimycotic activities were also tested on host plants and on three Seiridium species, virulent fungal agents of cypress canker disease. Some structure-activity relationships were identified for both phytoxicity and antifungal activities. These activities appear related to the presence of both pyridione and oxiran rings. Also the carbonyl group of the side chain seems to play a role into impart activity. PMID- 16712887 TI - Mini-symposium: health and environmental sustainability. The convergence of public health and sustainable development. PMID- 16712888 TI - Lack of aromatisation of the 3-keto-4-ene metabolite of tibolone to an estrogenic derivative. AB - Tibolone is used for the treatment of climacteric symptoms in postmenopausal women. It is metabolised in a tissue-specific manner so that while some metabolites exert estrogenic effects on bone and the CNS, others are thought to protect the breast and endometrium from estrogenic stimulation. Tibolone is a 7alpha-methyl derivative of 19-norethynodrel. Since the introduction of synthetic progestagens for therapeutic use there has been considerable controversy as to whether they can undergo aromatisation to give rise to the potent estrogen, ethinylestradiol. In this study, we examined whether the delta-4-ene (7alpha methyl norethisterone) metabolite of tibolone, which has a similar delta-4-ene A ring structure to that of the estrone precursor, androstenedione, could undergo aromatisation to the potent estrogen, 7alpha-methyl ethinylestradiol. For these studies, JEG-3 choriocarcinoma cells were employed as they have a very high level of aromatase activity. TLC and HPLC procedures were developed to separate phenolic from non-phenolic compounds and were initially used to confirm that JEG 3 cells readily aromatised androstenedione to estrogens (up to 74%). The aromatisation of androstenedione to estrogens by these cells could be completely blocked with the potent aromatase inhibitor letrozole. When [(3)H] 7alpha-methyl norethisterone was incubated with JEG-3 cells no evidence for its conversion to [(3)H] 7alpha-ethinylestradiol was obtained. Radioactivity detected on the TLC plate or HPLC fractions where standard 7alpha-methyl ethinylestradiol was located, revealed that similar levels were present when 7alpha-methyl norethisterone was incubated with culture medium alone or with JEG-3 cells in the absence or presence of letrozole. From these investigations, it is concluded that 7alpha-methyl norethisterone does not undergo aromatisation to an estrogenic derivative. PMID- 16712889 TI - Improving estimates of the basic reproductive ratio: using both the mean and the dispersal of transition times. AB - In both within-host and epidemiological models of pathogen dynamics, the basic reproductive ratio, R(0), is a powerful tool for gauging the risk associated with an emerging pathogen, or for estimating the magnitude of required control measures. Techniques for estimating R(0), either from incidence data or in-host clinical measures, often rely on estimates of mean transition times, that is, the mean time before recovery, death or quarantine occurs. In many cases, however, either data or intuition may provide additional information about the dispersal of these transition times about the mean, even if the precise form of the underlying probability distribution remains unknown. For example, we may know that recovery typically occurs within a few days of the mean recovery time. In this paper we elucidate common situations in which R(0) is sensitive to the dispersal of transition times about their respective means. We then provide simple correction factors that may be applied to improve estimates of R(0) when not only the mean but also the standard deviation of transition times out of the infectious state can be estimated. PMID- 16712890 TI - Histopathological effects induced by paraquat during Xenopus laevis primary myogenesis. AB - The oxidative agent paraquat induced tail abnormalities during Xenopus laevis development. Specimens exposed from blastula to the tadpole stage revealed pear shaped myocytes and irregular intersomitic boundaries. The histological feature of the axial musculature was evaluated in embryos sampled at significant stages of the primary myogenesis. During the somitogenesis PQ-treated embryos showed normal appearing myotomes, but reduced PAS activity in the post-rotating myotomal cells, and myoblasts with slight vacuolations. Once etched from the vitelline envelope, embryos showed severely altered myoblasts with irregular cellular apexes, heavy sarcoplasmic vacuolations, pyknotic nuclei and disorganizing intersomitic boundaries. Myotomes with many necrotic myocytes containing disorganized contractile material and heavily malformed intersomitic boundaries characterized the late myogenic stages. Our results evidence the heaviest PQ histopathological effects to affect myogenesis of post-etched embryos, suggesting a possible linkage between the swimming activity and the oxidative damage to muscle tissue. PMID- 16712891 TI - atRA-induced apoptosis of mouse embryonic palate mesenchymal cells involves activation of MAPK pathway. AB - Our previous studies have shown that atRA treatment resulted in cell-cycle block and growth inhibition in mouse embryonic palatal mesenchymal (MEPM). In the current study, gestation day (GD) 13 MEPM cells were used to test the hypothesis that the growth inhibition by atRA is due to apoptosis. The effects of atRA on apoptosis were assessed by performing MTT assay, Cell Death Detection ELISA and flow cytometry, respectively. Data analysis confirmed that atRA treatment induced apoptosis-like cell death, as shown by decreased cell viability and increased fragmented DNA and sub-G1 fraction. atRA-induced apoptosis was associated with upregulation of bcl-2, translocation of bax protein to the mitochondria from the cytosol, activation of caspase-3 and cytochrome c release into cytosol. atRA induced apoptosis was abrogated by z-DEVD-fmk, a caspase-3 specific inhibitor, and z-VAD-fmk, a general caspase inhibitor, suggesting that the atRA-induced cell death of MEPM cells occurs through the cytochrome c- and caspase-3-dependent pathways. In addition, atRA treatment caused a strong and sustained activation of c-Jun N-terminal kinase (JNK) and p38 kinase (p38), as well as an early but transient activation of extracellular signal-regulated kinase (ERK). Importantly, atRA-induced DNA fragmentation and capase-3 activation were prevented by pretreatment with the JNK inhibitor (SP600125) and the p38 MAPK inhibitor (SB202190), but not by pretreatment with MEK inhibitor (U0126). From these results, we suggest that mitogen-activated protein kinase-dependent pathways is involved in the atRA-induced apoptosis of MEPM cells. PMID- 16712892 TI - The regulation of cytochrome P450 2E1 during LPS-induced inflammation in the rat. AB - It is well known that inflammatory and infectious conditions differentially regulate cytochrome P450 (P450)-mediated drug metabolism in the liver. We have previously outlined a potential pathway for the downregulation in hepatic cytochrome P450 following LPS-mediated inflammation in the CNS (Abdulla, D., Goralski, K.B., Garcia Del Busto Cano, E., Renton, K.W., 2005. The signal transduction pathways involved in hepatic cytochrome P450 regulation in the rat during an LPS-induced model of CNS inflammation. Drug Metab. Dispos). The purpose of this study was to outline the effects of LPS-induced peripheral and central nervous system inflammation on hepatic cytochrome P450 2E1 (CYP2E1) in vivo, an enzyme that plays an important role in various physiological and pathological states. We report an increase in hepatic mRNA expression of CYP2E1 that occurred as early as 2-3 h following either the intraperitoneal (i.p.) injection of 5 mg/kg LPS or i.c.v. administration of 25 mug of LPS. This increase in CYP2E1 mRNA expression was sustained for 24 h. In sharp contrast to the increase in hepatic CYP2E1 mRNA, we observed a significant reduction in the catalytic activity of this enzyme 24 h following either the i.c.v. or i.p. administration of LPS. Cycloheximide or actinomycin-D did not change the LPS-mediated downregulation in hepatic CYP2E1 catalytic activity. Our results support the idea that LPS acts at two different levels to regulate hepatic CYP2E1: a transcriptional level to increase CYP2E1 mRNA expression and a post-transcriptional level to regulate CYP2E1 protein and activity. PMID- 16712893 TI - The farnesyltransferase inhibitor, LB42708, inhibits growth and induces apoptosis irreversibly in H-ras and K-ras-transformed rat intestinal epithelial cells. AB - LB42708 (LB7) and LB42908 (LB9) are pyrrole-based orally active farnesyltransferase inhibitors (FTIs) that have similar structures. The in vitro potencies of these compounds against FTase and GGTase I are remarkably similar, and yet they display different activity in apoptosis induction and morphological reversion of ras-transformed rat intestinal epithelial (RIE) cells. Both FTIs induced cell death despite K-ras prenylation, implying the participation of Ras independent mechanism(s). Growth inhibition by these two FTIs was accompanied by G1 and G2/M cell cycle arrests in H-ras and K-ras-transformed RIE cells, respectively. We identified three key markers, p21(CIP1/WAF1), RhoB and EGFR, that can explain the differences in the molecular mechanism of action between two FTIs. Only LB7 induced the upregulation of p21(CIP1/WAF1) and RhoB above the basal level that led to the cell cycle arrest and to distinct morphological alterations of ras-transformed RIE cells. Both FTIs successfully inhibited the ERK and activated JNK in RIE/K-ras cells. While the addition of conditioned medium from RIE/K-ras reversed the growth inhibition of ras-transformed RIE cells by LB9, it failed to overcome the growth inhibitory effect of LB7 in both H-ras- and K-ras-transformed RIE cells. We found that LB7, but not LB9, decreased the expression of EGFRs that confers the cellular unresponsiveness to EGFR ligands. These results suggest that LB7 causes the induction of p21(CIP1/WAF1) and RhoB and downregulation of EGFR that may serve as critical steps in the mechanism by which FTIs trigger irreversible inhibitions on the cell growth and apoptosis in ras-transformed cells. PMID- 16712894 TI - Enhanced urinary bladder and liver carcinogenesis in male CD1 mice exposed to transplacental inorganic arsenic and postnatal diethylstilbestrol or tamoxifen. AB - Pregnant CD1 mice received 85 ppm arsenite in the drinking water from gestation day 8 to 18, groups (n = 35) of male offspring were subsequently injected on postpartum days 1 through 5 with diethylstilbestrol (DES; 2 microg/pup/day) or tamoxifen (TAM; 10 microg/pup/day), and tumor formation was assessed over 90 weeks. Arsenic alone increased hepatocellular carcinoma (14%), adenoma (23%) and total tumors (31%) compared to control (0, 2 and 2%, respectively). Arsenic alone also increased lung adenocarcinoma, adrenal cortical adenoma and renal cystic tubular hyperplasia compared to control. Compared to arsenic alone, arsenic plus DES increased liver tumor incidence in mice at risk 2.2-fold and increased liver tumor multiplicity (tumors/liver) 1.8-fold. The treatments alone did not impact urinary bladder carcinogenesis, but arsenic plus TAM significantly increased formation of urinary bladder transitional cell tumors (papilloma and carcinoma; 13%) compared to control (0%). Urinary bladder proliferative lesions (combined tumors and hyperplasia) were also increased by arsenic plus TAM (40%) or arsenic plus DES (43%) compared to control (0%) or the treatments alone. Urinary bladder proliferative lesions occurred in the absence of any evidence of uroepithelial cytotoxic lesions. Urinary bladder lesions and hepatocellular carcinoma induced by arsenic plus TAM and/or DES overexpressed estrogen receptor-alpha, indicating that aberrant estrogen signaling may have been a factor in the enhanced carcinogenic response. Thus, in male CD1 mice, gestational arsenic exposure alone induced liver adenoma and carcinoma, lung adenocarcinoma, adrenal adenoma and renal cystic hyperplasia. Furthermore, DES enhanced transplacental arsenic induced hepatocarcinogenesis. In utero arsenic also initiated urinary bladder tumor formation when followed by postnatal TAM and uroepithelial proliferative lesions when followed by TAM or DES. PMID- 16712896 TI - Primary CCR5 only using HIV-1 isolates does not accurately represent the in vivo replicating quasi-species. AB - Most HIV-1 isolates depend on CCR5 or CXCR4 to infect target cells, and efficient use of other coreceptors is rare. We cloned HIV-1 envelopes from virus at acute infection and found that most use CCR3 efficiently. This result contradicts prevailing data, suggesting that CCR3 usage is rare. We hypothesized that direct isolation into PBMC biases selection of viruses that use CCR5 and not CCR3. We therefore compared coreceptor use of isolates obtained by PBMC coculture with envelopes cloned directly from patient blood samples, which should represent actively replicating species. Viruses derived by cloning generally used CCR3 and CCR5 with equally efficiently. In contrast, we found that viruses isolated by PBMC coculture largely, or exclusively, used CCR5. Regardless of whether CCR3 use contributes to HIV-1 transmission or pathogenesis, our results demonstrate that "primary isolates" generated by PBMC culture are unlikely to accurately represent the in vivo replicating quasi-species. PMID- 16712895 TI - Different European-type vaccines against porcine reproductive and respiratory syndrome virus have different immunological properties and confer different protection to pigs. AB - Immunization of piglets with two different European-type modified live vaccines against porcine reproductive and respiratory syndrome (PRRS) virus produced different outcomes. After vaccination, pigs became viremic (42 days), neutralizing antibodies did not develop, and frequencies of virus-specific gamma interferon-secreting cells (IFN-gamma-SC) were low. Levels of interleukin-10 (IL 10) produced by peripheral blood mononuclear cells (PBMC) seemed to inversely correlate with interferon-gamma responses. After a challenge with a virulent Spanish strain, one vaccine (V3) protected piglets against viremia while the other (V1) did not. The vaccine V3 induced the highest IFN-gamma-SC frequencies. IL-2, IL-4 or transforming growth factor-beta responses were not detected at any time for neither of the vaccines. In contrast, haptoglobin rose in sera of viremic pigs after the challenge. These results indicated a strong involvement of IFN-gamma, and maybe IL-10, in the development of immunity against PRRS virus. PMID- 16712898 TI - Author's reply to comment on "Biological removal of polychlorinated dibenzo-p dioxins from incinerator fly ash by sphingomonas wittichii RW1" by Rolf U. Halden. PMID- 16712897 TI - Production and characterization of vaccines based on flaviviruses defective in replication. AB - To develop new vaccine candidates for flavivirus infections, we have engineered two flaviviruses, yellow fever virus (YFV) and West Nile virus (WNV), that are deficient in replication. These defective pseudoinfectious viruses (PIVs) lack a functional copy of the capsid (C) gene in their genomes and are incapable of causing spreading infection upon infection of cells both in vivo and in vitro. However, they produce extracellular E protein in form of secreted subviral particles (SVPs) that are known to be an effective immunogen. PIVs can be efficiently propagated in trans-complementing cell lines making high levels of C or all three viral structural proteins. PIVs derived from YFV and WNV, demonstrated very high safety and immunization produced high levels of neutralizing antibodies and protective immune response. Such defective flaviviruses can be produced in large scale under low biocontainment conditions and should be useful for diagnostic or vaccine applications. PMID- 16712899 TI - Structure-activity relationship studies of resveratrol and its analogues by the reaction kinetics of low density lipoprotein peroxidation. AB - Resveratrol (3,5,4'-trans-trihydroxystibene) is a natural phytoalexin present in grapes and red wine, which possesses a variety of biological activities including antioxidative activity. To find more active antioxidants, with resveratrol as the lead compound, we synthesized resveratrol analogues, i.e., 3,4,3',4'-tetrahydroxy trans-stilbene (3,4,3',4'-THS), 3,4,4'-trihydroxy-trans-stilbene (3,4,4'-THS), 2,4,4'-trihydroxy-trans-stilbene (2,4,4'-THS), 3,3'-dimethoxy-4,4'-dihydroxy trans-stilbene (3,3'-DM-4,4'-DHS), 3,4-dihydroxy-trans-stilbene (3,4-DHS), 4,4' dihydroxy-trans-stilbene (4,4'-DHS), 3,5-dihydroxy-trans-stilbene (3,5-DHS) and 2,4-dihydroxy-trans-stilbene (2,4-DHS). Antioxidative effects of resveratrol and its analogues against free-radical-induced peroxidation of human low density lipoprotein (LDL) were studied. The peroxidation was initiated either by a water soluble initiator 2,2'-azobis(2-amidinopropane hydrochloride) (AAPH), or by cupric ion (Cu(2+)). The reaction kinetics were monitored either by the uptake of oxygen and the depletion of alpha-tocopherol (TOH) presented in the native LDL, or by the formation of thiobarbituric acid reactive substances (TBARS). Kinetic analysis of the antioxidation process demonstrates that these trans-stilbene derivatives are effective antioxidants against both AAPH- and Cu(2+)-induced LDL peroxidation with the activity sequence of 3,4,3',4'-THS approximately 3,3'-DM 4,4'-DHS>3,4-DHS approximately 3,4,4'-THS>2,4,4'-THS>resveratrol approximately 3,5-DHS>4,4'-DHS approximately 2,4-HS, and 3,4,3',4'-THS approximately 3,4-DHS approximately 3,4,4'-THS>3,3'-DM-4,4'-DHS>4,4'-DHS>resveratrol approximately 2,4 HS>2,4,4'-THS approximately 3,5-DHS, respectively. Molecules bearing ortho dihydroxyl or 4-hydroxy-3-methoxyl groups possess significantly higher antioxidant activity than those bearing no such functionalities. PMID- 16712900 TI - Hydrogen transfer pathways of the asymmetric reduction of alpha,beta-unsaturated ketone mediated by baker's yeast. AB - The hydrogen transfer mechanism of cofactor reduction and recycling processes in the yeast reduction of alpha,beta-unsaturated ketone was studied by using quantitative isotope tracing close to natural abundance measured by (2)H NMR. In the reaction, the active cofactor is NADPH. The cofactor-transferred hydride attacks the beta sp(2) carbon of the enone carbonyl while water hydrogen is transferred to the alpha position. The reductant involved in the reaction depends on the quantity of yeast. When the amount of yeast is very large, the enzymes use preferentially certain unidentified substance stored in the yeast cells rather than the added glucose as electron donor. In this case, the hydrogen transferred by the cofactor is mainly of water origin. When the yeast amount is low, the added glucose is more efficiently used by the enzymes as electron donor and its hydrogen atoms bound to C-1 and C-3 are delivered to the substrate. PMID- 16712901 TI - Aqueous salting-out effect of inorganic cations and anions on non-electrolytes. AB - The salting-out effects of 27 lithium, sodium, potassium, ammonium and magnesium salts and HCl on chloroform, benzene, chlorobenzene and anisole were characterized in aqueous solutions at 303 K by measuring the Henry's law constants. The concentration of the salt solutions was 0.5 mol dm(-3), i.e., similar to the salinity of sea water. The solubility change was described in terms of the Setschenow constant, K(S)(salt,solute). The highest salting-out effects were observed for the solutions of salts involving doubly charged anions, and the smallest for NO(-)(3). The individual ionic Setschenow constants, K(S)(cation,solute) and K(S)(anion,solute), were determined by multilinear regression, using the assumption of additivity for the ions. Cl(-) was selected as the reference ion for calculation of the K(S)(ion,solute) values of the other ions. The estimations resulted systematically in significant positive K(S)(cation,solute) values, ranging from 0.13+/-0.026 (NH(+)(4)) to 0.28+/-0.032 (Mg(2+)), which were hardly affected by the accompanying anion in solution, and only slightly affected by the non-electrolytes present. NO(-)(3) resulted in a slight salting-in effect: K(S)(NO(-)(3),solute)=-0.083+/-0.019; the other anions displayed salting-out effect for all of the non-electrolytes studied, with K(S)(anion,solute) ranging between 0.090+/-0.008 (HCO(-)(3)) and 0.21+/-0.035 (CO(2-)(3)). PMID- 16712902 TI - On the critique of flake recovery rate and behavioral inference for early hominids: a reply to. PMID- 16712903 TI - Size and shape variation in Australopithecus afarensis proximal femora. AB - The degree of size and shape variation in the A. afarensis fossil sample has been interpreted in a variety of ways. Size variation has been described as exceeding that of extant hominoids, similar to that of strongly sexually dimorphic hominoids, and best matched to modern humans. The degree of shape variation has been characterized both as great and negligible. Recent fieldwork has increased the proximal femoral sample, providing new data with which to examine variation. The proximal femur of A. afarensis is analyzed in a comparative framework in order to gauge the magnitude of size and shape variation in this element. Seven of the best-preserved A. afarensis proximal femora contribute to the analysis (A.L. 128-1, A.L. 152-2, A.L. 211-1, A.L. 288-1ap, A.L. 333-3, A.L. 333-123, A.L. 827-1). Comparative samples from Pan, Pongo, Gorilla, and Homo provide context for interpreting variation among the fossils. The coefficient of variation (CV) of linear measurements is used to estimate size variation. Bootstrap resampling of CVs from extant hominoids provides distributions for comparison to A. afarensis CVs. Ratios of linear measurements provide scale-free shape variables that are used in pairwise comparisons. The Euclidean distance between pairs of A. afarensis are compared to the Euclidean distances between extant hominoid pairs. As found in some earlier analyses, size variation in A. afarensis is accommodated best in gorillas and orangutans. The magnitude of difference in shape between A. afarensis pairs is exceeded by most taxa, indicating that shape variation is not extreme. These general findings are contradicted by a few instances of excessive size and shape variation. These are uncharacteristic results and could point to temporal bias, although other alternatives are explored. The signal from the proximal femur is that size variation in A. afarensis is like that of the strongly sexually dimorphic apes, and shape variation is well within the range of most hominoids irrespective of their degree of size dimorphism. PMID- 16712904 TI - The respective and combined anticoagulant effects of recombinant human activated protein C, melagatran and heparins using CAT. AB - INTRODUCTION: Combinations of anticoagulants might be beneficial in some patients with sepsis, but most anticoagulants require specific clotting assays for monitoring. Thrombin generation assay, however, is a global function test of hemostasis. MATERIALS AND METHODS: We performed an in vitro investigation of the respective effects of recombinant human activated protein C (rhAPC) alone and in combination with either melagatran (a new direct thrombin inhibitor), unfractionated heparin (UH) or low molecular weight heparin (LMWH) in varying concentrations on the thrombin generation (TG) using the calibrated automated thrombography. RESULTS: RhAPC, UH, LMWH and melagatran dose-dependently prolonged the lag time and the time to peak, and significantly suppressed the endogenous thrombin potential (ETP). Combined application of rhAPC with either melagatran, UH or LMWH induced an additive prolongation of the lag time; this effect was more pronounced in a combination of rhAPC with UH or LMWH. CONCLUSION: In our in vitro study adding either melagatran, UH or LMWH augmented the capacity of rhAPC to suppress thrombin generation in human plasma. These findings suggest that patients with severe sepsis might benefit from a treatment with combinations of anticoagulant agents. PMID- 16712905 TI - Phase II study of oxaliplatin as second-line chemotherapy in endometrial carcinoma: a Gynecologic Oncology Group study. AB - OBJECTIVE: A phase II study was conducted to determine the efficacy of oxaliplatin therapy in patients with advanced or recurrent endometrial cancer who had received one prior platinum therapy. METHODS: Eligible patients were to have measurable disease and one prior chemotherapy regimen which could include cisplatin or carboplatin. Oxaliplatin 130 mg/m2 was administered intravenously over 2 h. This treatment was repeated every 21 days until progression of disease or adverse effects prohibited further therapy. RESULTS: Fifty-four patients were entered on study, of which 52 were eligible and 50 had had prior platinum therapy. The overall response rate was 13.5%, with three patients (5.8%) achieving a complete response and four patients (7.7%) achieving a partial response. Median duration of response was 10.9+ (range: 4.1-50.3+) months. Stable disease was reported in 15 (28.8%) patients, with an associated median duration of 5.4 (range: 2.2-19.6) months. Drug-related toxicities consisted of anemia, nausea and vomiting, and neurotoxicity. CONCLUSIONS: Oxaliplatin at the dose and schedule employed has limited activity in patients with advanced or recurrent endometrial carcinoma who have had previous platinum therapy. PMID- 16712906 TI - Estrogen replacement therapy and tamoxifen are contraindicated in patients with endometrial stromal sarcoma. PMID- 16712907 TI - Absence of the BH3 domain mutation of proapoptotic Bcl-2 family gene BAD in serous and mucinous tumors of ovary. PMID- 16712908 TI - New insights regarding glucocorticoids, stress and gonadotropin suppression. AB - This review highlights our recent work investigating the inhibitory effects of acute, physiologic stress-like increases in cortisol on reproductive neuroendocrine activity in sheep, the mechanisms responsible for this suppression, and the relevance of enhanced glucocorticoid secretion to stress induced inhibition of gonadotropin secretion in this species. Initial studies established that cortisol rapidly suppresses pulsatile luteinizing hormone secretion. In ovariectomized ewes, this inhibition reflects the reduction of pituitary responsiveness to gonadotropin-releasing hormone mediated by the type II glucocorticoid receptor, rather than the suppression in hypothalamic gonadotropin-releasing hormone release. Studies in ovary-intact ewes, however, uncovered an alternative mode of cortisol action. During the follicular phase of the estrous cycle, cortisol reduces luteinizing hormone pulse frequency, most likely via the inhibition of gonadotropin-releasing hormone pulsatility. Recent preliminary evidence in ovariectomized ewes demonstrates increased cortisol secretion is essential for disruption of pulsatile luteinizing hormone secretion in response to a psychosocial stress. Taken together, our observations reveal diverse inhibitory actions of cortisol on gonadotropin secretion and that this glucocorticoid is not only sufficient, but necessary for suppression of reproductive neuroendocrine activity in response to certain types of stress. PMID- 16712909 TI - Chronic running wheel activity attenuates the antinociceptive actions of morphine and morphine-6-glucouronide administration into the periaqueductal gray in rats. AB - Chronic exercise in a running wheel increases baseline pain sensitivity while attenuating the antinociceptive effects of peripherally administered opiate agonists in laboratory rodents. To determine if these effects are due to exercise induced changes in the central nervous system (CNS) or an artifact of exercise induced alterations in peripheral physiology, the present study evaluated the antinociceptive actions of centrally administered opiate agonists in active and inactive female rats. Rats were implanted with cannula into the right periaqueductal gray (PAG) area of the midbrain. After the completion of the surgery, the animals were allowed ad libitum access to running wheels or housed in standard cages for three weeks. Pain sensitivity was measured on the tail flick test before and immediately following microinjections of either morphine (0, 2.5, 5.0, 10.0, 20.0 microg/rat) or the more potent morphine metabolite, morphine-6-glucuronide (M6G) (0, 0.03, 0.1, 0.3, 1.0 microg/rat). Baseline tail flick latencies were significantly shorter in active than in inactive rats. Additionally, active animals were less sensitive to the antinociceptive effects of morphine and M6G than inactive rats. These findings provide evidence for the involvement of the CNS in exercise-mediated alterations in pain sensitivity and opiate drug actions. PMID- 16712910 TI - The involvement of endogenous opioid mechanisms in the antinociceptive effects induced by antidepressant drugs, desipramine and trimipramine. AB - The present study was designed to investigate the involvement of endogenous opioid systems in the antinociception induced by the antidepressant drugs, desipramine and trimipramine. For this purpose, the antinociceptive effects of desipramine (7.5 and 15.0 mg/kg i.p.) and trimipramine (5.0 and 10.0 mg/kg i.p.) were compared to that induced by morphine (0.2 and 2.0 mg/kg i.p.) in the tail clip model in mice. Naloxone (0.3 and 3.0 mg/kg i.p.), a non-specific opioid receptor antagonist, inhibited morphine-induced antinociception in mice, whereas the antinociceptive effects of antidepressant drugs were found to be resistant to naloxone blockade to some extent, since only the higher concentration of naloxone (3.0 mg/kg i.p.) caused significant inhibition of the effects of antidepressant drugs. In contrast, naltrindole (1.0 mg/kg i.p.), a specific delta-receptor antagonist, inhibited antinociception induced by desipramine and trimipramine in this test, while it inhibited the antinociceptive effect of morphine only partly. None of the opioid antagonists produced a significant effect in the tail-clip experiment when they were injected alone. Based on these findings, we concluded that endogenous opioids are involved in the antinociceptive effects of the antidepressant drugs using different mechanisms. PMID- 16712912 TI - Parents' perceptions of neighborhood safety and children's physical activity. AB - OBJECTIVE: The obesity epidemic disproportionately affects minority and poor children. Negative perceptions of neighborhood safety in poor communities may affect overweight by inhibiting children's physical activity. This study investigates the degree to which parents in a poor inner city vs. a middle-class suburban community limit their children's outdoor activity because of neighborhood safety concerns. METHOD: Parents of children aged 5-10 years from an inner city family practice in a poor community and from a suburban pediatric practice in a middle-class community completed a 20-item questionnaire. Parents estimated the amount of their child's activity in various situations and indicated their level of anxiety concerning gangs, child aggression, crime, traffic, and personal safety in their neighborhood. RESULTS: Inner city children (n = 204) engaged in less physical activity than suburban children (N = 103) (P < 0.001). Inner city parents expressed much greater anxiety about neighborhood safety than suburban parents (P < 0.0001). In the inner city population, children's physical activity levels were negatively correlated with parental anxiety about neighborhood safety (r = -0.18, P < 0.05). CONCLUSIONS: Inner city parents have high levels of anxiety about neighborhood safety. While these concerns may not entirely explain the discrepancy in activity levels between inner city and suburban children, a safe environment is crucial to increasing opportunities for physical activity. PMID- 16712911 TI - Individual and family household smoking habits as risk factors for wheezing among adolescents. AB - BACKGROUND: The study aimed to evaluate the household smoking habits as risk factors for wheezing in adolescents. METHODS: During a 2-month period, the study was performed by applying the standardized ISAAC phase 1 questionnaire to 3474 adolescents attending a random sample of secondary public schools located at the southeast of Mexico City. Adolescents were grouped as "cases" if they had at least one episode of wheezing in the year prior to the study, or as "controls" if they did not. Positive individual and family (father, mother or other persons living in the home) smoking habits at home were evaluated as potential risk factors. RESULTS: More than 98% of respondents in the two groups reported that the father smoked in the house. More mothers who smoked were reported among cases than controls (OR 2.42; 95% CI 1.60-3.68). More adolescents in the group of cases had history of ever having rhinitis, rash or wheezing. Fifty-four percent of adolescents in the group of cases had ever smoked versus 38.8% of controls (OR 1.89; 95% CI 1.33-2.67). Sex-adjusted logistic regression analysis identified a "protective" role for wheezing when the number of smokers at home was < or =2 (OR 0.44; 95% CI 0.27-0.71). CONCLUSION: Individual, maternal and other family members' indoor smoking habits may increase the risk of wheezing among adolescents. PMID- 16712913 TI - How to simulate wear? Overview of existing methods. AB - OBJECTIVES: The aim of the overview is to critically analyse the contributing factors to the biological wear process and to screen the wear simulators in dentistry for their capacity to mimic the wear conditions. An overview of the types of wear, grouped as biotribocorrosion, combined with a description of the different wear simulating devices will allow us to better understand the multifactorial nature of wear. METHODS: A search on keywords highlights the most common in-vitro wear simulators and their use in the laboratories for various simulation applications. RESULTS: Wear is a complex process that can hardly be simulated while controlling all variables. Especially the extrapolation of the in vitro wear results to the in-vivo situation is difficult because there is a lot of interplay with biological factors that are difficult to mimic. SIGNIFICANCE: It is not the degree of sophistication, but the right mix of controllable variables that will make a wear simulator predictive. PMID- 16712914 TI - Family heritage and depression guides: family and peer views influence adolescent attitudes about depression. AB - While adolescents tend to under-use professional mental health services for depression, they informally seek health-related information from parents and peers. In this study, we interviewed 15 adolescents to examine how the views and behaviours of others influence teens' decisions about seeking care for depression. Using a grounded theory approach, we found that teens' families, peers, and siblings contributed uniquely to teens' decisions in seeking care for depression. Families may disclose a "heritage" of depression, and their choices about disclosing family mental health issues, previous treatment, and coping strategies affected teens' understanding of depression. Peer "depression guides," who had themselves experienced depression, provided teens with advice on recognizing depression, managing stigma, and seeking care. Siblings bridged the roles of peer and family, influencing teens' choices about accessing treatment. These findings suggest that family and peers should be included during teens' depression treatment. PMID- 16712915 TI - An exploratory study of the relationship of family support and coping with adjustment: Implications for college students with a chronic illness. AB - To examine the role of family support and coping in the adjustment of adolescents with chronic illness (CI) transitioning into college, college freshmen and sophomores (N(chronic illness)=32, N(primarily negative life event)=53, N(primarily positive life event)=16) were administered standard measures. CI group reported more anxiety and a trend towards more depression than the primarily positive life events group (PLE). Coping strategy (active and passive) was positively associated with anxiety in CI group, whereas, family support was negatively associated with depression and anxiety in the primarily negative life events group (NLE), and passive coping was positively associated with depression in PLE. Development of adaptive coping should be the focus of programs aimed at supporting students with CI as they transition to college. PMID- 16712916 TI - Towards structural biology in the genus Mytilus: on-line homology modeling with Geno3D. AB - Seven structures from the genus Mytilus exist in the protein data bank (PDB) but > 1000 Mytilus protein sequences are known. Sequences (NCBI) were used as targets for on-line homology modeling with Geno3D as an alternative route to structure. Quality comparators include root mean square deviation (RMSD) of target from template, chain geometry and Ramachandran diagram analysis. We modeled 17 Mytilus structures for mainly stress-response proteins relevant to biomonitoring. These model files are available as a first step towards a structural resource for Mytilus. Analysis suggests that they are structurally plausible with low RMSDs relative to targets. Ramachandran analysis suggests a small percentage of disallowed dihedral angles (in the range 0-6.86% for 15 of 17 models). PMID- 16712917 TI - Are low doses of tritium genotoxic to Mytilus edulis? AB - The genotoxic effects of tritium (3H) in the adult life stage of Mytilus edulis have been evaluated by the induction of micronuclei (MN) and DNA single strand breaks/alkali labile sites (Comet assay) in the haemocytes of exposed individuals. Assays were optimised and validated using ethylmethane sulfonate (EMS) as a reference genotoxic agent over different exposure periods. M. edulis were exposed, for 96 h, to a range of concentrations of 3H equivalent to a dose range of 12-485 microGy h(-1). Results revealed a dose-dependent increase for both the MN and Comet assays, and for both EMS and 3H. Since less than 500 microGy h(-1) 3H is capable of inducing genetic damage, generic doses recommended by the IAEA for the protection of aquatic biota may be overestimated for some organisms. PMID- 16712918 TI - Application of SSH and a macroarray to investigate altered gene expression in Mytilus edulis in response to exposure to benzo[a]pyrene. AB - The lack of genomic resources for aquatic invertebrates restricts their use as sentinel species in coastal environments. It is known that where genomic data are not available, suppression subtractive hybridisation (SSH) can generate cDNA libraries representative of pollutant-responsive gene transcription in aquatic vertebrates. To assess whether the approach was equally suited to aquatic invertebrates, altered gene expression in digestive gland of the mussel, Mytilus edulis, in response to exposure to benzo[a]pyrene (BaP) (1 mg/l) was investigated with SSH and a nylon macroarray. Screening of the subtracted libraries showed 112/250 up-regulated and 25/55 down-regulated clones were positive for differential expression and characterisation of these identified 87 with unique sequence suitable for array on a nylon membrane. The transcripts isolated were from a diverse range of genes involved in general stress, oxidative stress, cell adhesion, transcriptional and translational regulation, transport mechanisms, energy metabolism, cell metabolism, lipid metabolism, protein turnover and activation, lysosomal activity and 22 cryptic clones. Subsequent use of the clones in macroarray format to analyse expression of BaP-responsive genes (0 vs 4 day exposed) showed 0-100-fold increased levels of the forward-subtracted probes and between 0 and 0.1-fold down-regulation of the reverse-subtracted probes. Only 15% of the clones showed less than 2-fold change in expression. The gene ontology of the transcripts isolated demonstrates that BaP elicits a multitude of responses with a major feature being disruption of cellular redox status. The results indicate that the use of SSH and a macroarray is a robust method to discover novel pollutant-responsive genes in aquatic invertebrates. PMID- 16712919 TI - Anti-androgen flutamide affects gonadal development and reproduction in medaka (Oryzias latipes). AB - The effects of an anti-androgenic chemical on the reproduction of medaka (Oryzias latipes) were examined. Six mating pairs of medaka were exposed to flutamide (FLU) at measured concentrations of 0.101, 0.202, 0.397, 0.787, or 1.56 mg/L for 3 weeks. During the exposure period, one female died in the 1.56-mg/L treatment group, but no lesions or pathological abnormalities were observed. The fecundity and fertility of paired medaka were significantly decreased at 1.56 mg/L compared with those of the controls. Histological examination showed intersex gonads (testis-ova) in males exposed to FLU at 0.202, 0.397, and 0.787 mg/L. However, concentrations of hepatic vitellogenin (Vtg) in both sexes were not statistically different from those in the controls. These results demonstrate that FLU affected gonadal development in male medaka and reproductive capacity in paired medaka. This is the first report of testis-ova in male medaka exposed to FLU. PMID- 16712920 TI - Cloning and expression of MDR transporters from marine bivalves, and their potential use in biomonitoring. AB - Multidrug resistance transporters (MDRs) are excellent candidates for molecular level biomonitoring - they function in exporting xenobiotic compounds and their expression is inducible. However, currently available MDR sequence information from aquatic invertebrates is partial and mostly biased towards the conserved ATPase domain. In the present study, two genes belonging to the MDR/TAP (ABCB) family were cloned and characterized from the bivalve Brachidontes pharaonis, which thrives in rocky environments along the Israeli Mediterranean coast. One of these is a complete sequence of a 'half'ABCB, probably belonging to the ABCB10 subfamily, while the second is a 'full'ABCB1 transporter. A quantitative RT-PCR protocol for biomonitoring was tested in laboratory experiments. Bivalves exposed to diesel showed significant increase in B1 expression levels, while the expression of B10 was suppressed. These results suggest that B. pharaonis features an MDR1 homologue that is induced by pollution and may serve as a sentinel organism for routine biomonitoring programs. However, our findings also exemplify that not all MDRs are equally suitable for this purpose and sequence information must be expanded beyond the ATPase domain for correct classification of cloned genes. PMID- 16712921 TI - A realistic evaluation of the action of ozone on whole human blood. AB - We have clarified the role of the ozone concentration in relation to the resistance of human erythrocytes in whole human blood or in blood diluted either in saline or in distilled water. Spectrophotometric data related to haemoglobin were evaluated by exposing samples of fresh human blood directly to ozone doses (ratio 1:1 volume), within the therapeutic range (0.21-1.68 mM) and to one toxic dose (3.36 mM). Furthermore, the same determinations have been carried out after previous dilution of the same blood with either pure water or physiological saline (1 ml blood + 29 ml diluent) followed by ozonation with the above reported ozone doses. Addition of either saline or water implies a dilution of plasma antioxidants and also total haemolysis after water dilution. Particularly the latter case represents a most unphysiological situation because the osmotic shock causes the solubilization of the erythrocytic content. While it is possible to demonstrate that after haemolysis there is an ozone-concentration dependent transformation of some oxyhaemoglobin to methaemoglobin, no such process occurs after ozonation of whole blood. The results of this study fully confirm our previous data that judicious ozone doses neither damage erythrocytes, nor induce oxidation of intracellular haemoglobin. We hope that our conclusions will definitively clarify the absence of toxicity of ozonetherapy. PMID- 16712922 TI - Effect of acetonitrile on Cynara cardunculus L. cardosin A stability. AB - The kinetics of the structural changes affecting cardosin A, a plant aspartic proteinase (AP) from Cynara cardunculus L., in the presence of a mixture of acetonitrile (AN) in water (W) was studied. Incubation of cardosin A with 10% (v/v) AN resulted in a gradual increase in protein helicity, accompanied by changes in the tertiary structure, seen by changes in the intrinsic fluorescence of tryptophan. Differential scanning calorimetry (DSC) revealed that the temperature of denaturation of cardosin A decreased upon the addition of AN. With longer incubation times, the small chain of cardosin A denatured completely, consequent exposure of the single tryptophan residue accounting well for the observed spectral shift intrinsic fluorescence of the protein. Enzymatic activity assays demonstrated that the kinetically determined unfolding of the small chain of cardosin A does not result in loss of the activity of this enzyme. PMID- 16712923 TI - An evidence for the equilibrium unfolding intermediates of ribonuclease A by tritium labeling method. AB - A formation of a molten globule in the unfolding of ribonuclease A could be considered as an evidence supporting a hypothesis on the existence of such intermediates on the pathway of a protein folding. Using a novel technique (tritium labeling method) we have showed that the ribonuclease A equilibrium unfolding in urea and guanidinium chloride (GuCl) solutions proceeds through a formation of intermediates whose properties (compactness, retention of the larger part hydrophobic core, secondary structure, and native-like folding pattern) correspond to the fundamental characteristics of the molten globule state. The both intermediates are the "wet" molten globules (the globule interior contains the water molecules). The results reveal the noticeable distinctions in intermediates structure, first of all, in the extent of their compactness. The urea intermediate is less compact than that in GuCl. It is shown that the refolding of the protein denatured by GuCl results in the formation of the intermediate which enzyme activity is virtually the same as the activity of the native protein. PMID- 16712924 TI - Trypsin immobilization by direct adsorption on metal ion chelated macroporous chitosan-silica gel beads. AB - Silica gel bead coated with macroporous chitosan layer (CTS-SiO(2)) was prepared, and the metal immobilized affinity chromatographic (IMAC) adsorbents could be obtained by chelating Cu(2+), Zn(2+), Ni(2+) ions, respectively on CTS-SiO(2), and trypsin could be adsorbed on the IMAC adsorbent through metal-protein interaction forces. Batch adsorption experiments show that adsorption capacity for trypsin on these IMAC adsorbent variated with change of pH. The maximal adsorption reached when the solution was in near neutral pH in all three IMAC adsorbents. Adsorption isothermal curve indicated that maximal adsorption capacity could be found in the Cu(2+)-CTS-SiO(2) with the value of 4980+/-125 IUg(-1) of the adsorbent, while the maximal adsorption capacity for trypsin on Zn(2+) and Ni(2+) loaded adsorbent was 3762+/-68 IUg(-1) and 2636+/-53 IUg(-1), respectively. Trypsin immobilized on the IMAC beads could not be desorbed by water, buffer and salt solution if the pH was kept in the range of 5-10, and could be easily desorbed from the IMAC beads by acidic solution and metal chelating species such as EDTA and imidazole. The effect of chelated metal ions species on CTS-SiO(2) beads on the activity and stability of immobilized trypsin was also evaluated and discussed. Trypsin adsorbed on Zn-IMAC beads retained highest amount of activity, about 78% of total activity could be retained. Although the Cu-IMAC showed highest affinity for trypsin, only 25.4% of the calculated activity was found on the beads, while the activity recovery found on Ni-IMAC beads was about 37.1%. A remarkable difference on stability of trypsin immobilized on three kinds of metal ion chelated beads during storage period was also found. Activity of trypsin on Cu-IMAC decreased to 24% of its initial activity after 1-week storage at 4 degrees C, while about 80% activity was retained on both Ni-IMAC and Zn-IMAC beads. Trypsin immobilized on Zn-CTS-SiO(2) could effectively digest BSA revealed by HPLC peptide mapping. PMID- 16712925 TI - Synthesis and characterization of pseudo-affinity ligand for penicillin acylase purification. AB - The aim of this work was to test a chromatographic affinity support containing methacryloyl antipyrine (MAAP) for penicillin acylase (PA) purification by using pure penicillin acylase and crude extract. First, MAAP as a pseudo-specific ligand was synthesized by using methacryloyl chloride and 4-aminoantipyrine. Polymer beads (average size diameter: 40-120 micro m) were prepared by suspension polymerization of ethylene glycol dimethacrylate (EGDMA) and MAAP. This approach for the preparation of adsorbent has several advantages over conventional preparation protocols. An expensive and time consuming step in the preparation of adsorbent is immobilization of a ligand to the adsorption matrix. In this procedure, affinity ligand MAAP acts as comonomer without further modification steps. Poly(EGDMA-MAAP) beads were characterized by FTIR, NMR and screen analysis. Elemental analysis of MAAP for nitrogen was estimated as 89.3 micro mol/g. The prepared adsorbent was then used for the capture of penicillin acylase in batch system. The maximum penicillin acylase adsorption capacity of the poly(EGDMA-MAAP) beads was found to be 82.2 mg/g at pH 5.0. Chromatography with crude feedstock resulted in 23.2-fold purification and 93% recovery with 1.0 M NaOH. PMID- 16712926 TI - The effects of combined gentamicin-hydroxyapatite coating for cementless joint prostheses on the reduction of infection rates in a rabbit infection prophylaxis model. AB - Infections remain a critical issue in total joint arthroplasty. Addition of antibiotics to bone cement was shown to significantly improve antimicrobial prophylaxis in cemented joint arthroplasty. In cementless joint arthroplasty a comparable prophylaxis by local antibiotics has not been possible yet. The aim of the current study was to investigate the antimicrobial effect of two different gentamicin-hydroxyapatite (HA) coatings for cementless prostheses in a rabbit infection model. Staphylococcus aureus with a dose of 10(7) CFUs was inoculated into the intramedullary canal of the tibia of 30 rabbits followed by the implantation of standard steel HA K-wires (n=10), steel K-wires coated with a gentamicin-HA combination (n=10), and steel K-wires coated with a gentamicin-RGD HA combination (n=10), respectively. The animals were sacrificed after 28 days and clinical, histological and microbiological assessment on the bone and on the removed K-wire itself by agar plating and DNA-pulsed field gel electrophoresis were carried out to detect infection. There was a statistically significant reduction of infection rates by both gentamicin-coating types (0 infections in both groups) compared to standard HA coating (7 infections in 8 animals; 2 animals were lost due to acute diarrhea) (p<0.001). An excellent correlation between agar plating testing results of the K-wires and of the bone samples was found. Detailed histology showed cortical lysis, abscess and sequester formation in the infected animals. Both gentamicin-coating types showed significant improvement of infection prophylaxis compared to standard HA coating and, therefore, this coating technology could help to improve infection prophylaxis in cementless total joint arthroplasty. In further studies biocompatibility of the coatings has to be assessed. PMID- 16712927 TI - Evolutionary differentiation of Cetartiodactyl placentae in the light of the viviparity-driven conflict hypothesis. AB - We analysed the evolution of placental traits in the novel mammalian clade Cetartiodactyla (Cetaceans and Artiodactyls) by a parsimony-based computer program (MacClade). A diffuse epitheliochorial placenta was identified as the stem species pattern of this clade. Trophoblast giant cells (TGCs) independently evolved in Camelids and Ruminants. The polycotyledonary placenta is an apomorphic character for Pecora (higher ruminants) and the oligocotyledonary placenta developed as a further step on the stem lineage of cervidae and moschidae. We interpret these findings by application of the "viviparity-driven conflict hypothesis", which states that divergent interests of mother and offspring lead to a rapid antagonistic coevolution, which might cause placental diversity. According to this hypothesis the evolution of camelid and ruminant TGCs can be interpreted as means to increase fetal endocrine influence on the maternal metabolism. The development of the cotyledonary placenta could be related to a diminished availability of glucose, which is associated with the evolution of forestomach fermentation in Pecora. An arms race, in which the mother tried to restrict and the fetus tried to increase transplacental glucose flow, might have promoted the evolution of the cotyledonary placenta, which has a high feto maternal exchange area, but a low conductivity for glucose. PMID- 16712928 TI - Expression in human trophoblast and choriocarcinoma cell lines, BeWo, Jeg-3 and JAr of genes involved in the hepatobiliary-like excretory function of the placenta. AB - Using cytokeratin-7-positive trophoblast cells (hTr) isolated from human term placentas and the choriocarcinoma cell lines (hCC) BeWo, Jeg-3 and JAr, the expression of genes involved in the hepatobiliary excretion of cholephilic compounds was investigated by RT-PCR/sequencing followed by measurement of the absolute abundance of mRNA by real-time RT-PCR. Although mRNA of BSEP was detectable and its expression confirmed by Western blotting, its very low expression (higher in hTr than in whole placenta and hCC) did not permit its detection by immunohistochemistry. In hTr, the expression was high for OATP B/2B1, OATP-8/1B3, MRP1, MRP3, BCRP, FIC1, RARalpha, FXR and SHP, low for OSTalpha, MRP2, MRP4, MRP8, MDR1, CAR and SXR, very low for OATP-A/1A2 and MDR3, and not detectable for OATP-C/1B1, HNF1alpha and HNF4. Expression patterns in hCC mimicked those in hTr, although some important cell line-specific differences were found. The functionality of transporters expressed in hCC was confirmed by their ability to take up and export estradiol 17beta-d-glucuronide in a self inhibitable and temperature-sensitive manner. In conclusion, several transporters, export pumps, and nuclear receptors involved in the liver excretory function may play a similar role in the placenta, whose specific aspects can be studied by selectively using BeWo, Jeg-3 or JAr cells. PMID- 16712929 TI - Synthetic bovine proline-rich-polypeptides generate hydroxyl radicals and fail to protect dopaminergic neurons against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine induced dopaminergic neurotoxicity in mice. AB - Proline-rich-polypeptides (PRPs) isolated from bovine hypothalamus have been shown to render protection against neuronal injury of the brain and spinal cord. We examined two PRPs containing 15 and 10 amino acid residues (PRP-1 and PRP-4 synthetic polypeptide) for their effect, if any, on dopaminergic neuronal damage caused by the parkinsonian neurotoxin, 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP). Effects of these PRPs on hydroxyl radical ((*)OH) generation in a Fenton-like reaction as well as from isolated mitochondria were monitored, employing a sensitive salicylate hydroxylation procedure. Balb/c mice treated (i.p., twice, 16 h apart) with MPTP (30 mg/kg) or PRP-1 (1.6 mg/kg), but not PRP-4 (1.6 mg/kg) showed significant loss of striatal dopamine and norepinephrine as assayed by an HPLC-electrochemical procedure. Pretreatment with the PRPs, 30 min prior to the neurotoxin administration failed to attenuate MPTP induced striatal dopamine or norepinephrine depletion, but significantly attenuated the MPTP-induced decrease in dopamine turnover. A significant increase in the generation of (*)OH by the PRPs in a Fenton-like reaction or from isolated mitochondria suggests their pro-oxidant action, and explains their failure to protect against MPTP-induced parkinsonism in mice. PMID- 16712930 TI - Hypermethylation of CpG island in the promoter region of CALCA in acute lymphoblastic leukemia with central nervous system (CNS) infiltration correlates with poorer prognosis. AB - Promoter hypermethylation occurs early in leukemogenesis and seems to be associated with poor prognosis in acute lymphoblastic leukemia (ALL). The methylation status of the promoter region of six genes was analyzed in 71 children with ALL using methylation specific PCR (MSP). Calcitonin (CALCA) and E cadherin (CDH1) were the most frequently methylated genes in this group of patients. Considering the patients with central nervous system (CNS) infiltration, the estimated 2-year overall survival (OS) was 20% for those with methylation in CALCA promoter and 85% for those without (p=0.001). Our results suggest that the hypermethylation of CALCA promoter is a promising prognostic marker and may predict a higher risk for ALL patients with CNS infiltration. PMID- 16712931 TI - Antecedents and concomitants of parenting stress in adolescent mothers in foster care. AB - OBJECTIVE: This study's aim was to examine variables associated with different short-term trajectories in multiply disadvantaged adolescent mothers by investigating antecedents and concomitants of parenting stress. METHOD: We followed 49 adolescent mothers (ages 14-18 at study outset) who were wards in Illinois foster care using a longitudinal correlational design. We examined whether parenting variables (childrearing beliefs, quality of parent-child interactions, and child abuse risk) and personal adjustment variables (emotional distress and social support) at initial assessment predicted parenting stress measured at follow-up (a mean of 22.5 months later). We also examined concurrent relationships between parenting stress and mothers' adaptive functioning in educational, social support, and childbirth areas at follow-up. RESULTS: We found that parenting variables, but not personal adjustment variables, predicted later parenting stress. Results also showed that current adaptive functioning was significantly related to parenting stress. Specifically, educational status and social support predicted concurrent parenting stress, whereas number of childbirths did not. CONCLUSIONS: These findings extend the small literature on the link between parenting difficulties and parenting stress to adolescent mothers in foster care. Parenting challenges, particularly as reflected in unrealistic childrearing expectations, appear to be markers for later parenting stress. Considering the longitudinal relationships observed, early and periodic assessment of adolescent mothers' parenting knowledge, skills, and interactions is recommended. Also, given that this study found concurrent social support and educational status to covary with current parental stress, these variables, and others for which they may serve as proxy, are implicated for careful monitoring. PMID- 16712932 TI - Death by hypnosis: an 1894 Hungarian case and its European reverberations. AB - The story of a fatal hypnotic seance in a castle in provincial Hungary in 1894 was sensationalised by the media and propelled across national and social boundaries within a few days. It stirred public feelings and compelled prestigious medical mandarins, legal professionals and social commentators of the day to express wide-ranging views concerning hypnotic practice. The case intensified social and professional anxieties surrounding hypnosis in late 19th century culture and illustrates the complex relationship between medical hypnotic research, lay hypnosis and widely reported and sensationalised forensic cases. PMID- 16712933 TI - Cloning and characterization of guinea pig CXCR1. AB - IL-8/CXCL8 plays a critical role in the trafficking and activation of neutrophils via its receptors, CXCR1 and CXCR2, in humans. CXCR1 is highly selective for IL 8, whereas CXCR2 is activated by all CXC chemokines with an ELR motif. In mice and rats, neither IL-8 nor CXCR1 is present, making it difficult to evaluate the in vivo roles of the IL-8/CXCR1 interactions. We previously demonstrated the presence of IL-8 in the guinea pig (gp), suggesting that its specific receptor CXCR1 is also present in this species. Here, we obtained two gp genomic DNA clones, clones 8 and 10, coding for the potential orthologues of CXCR1 and CXCR2, respectively. Transcripts for these genes were expressed in neutrophils, but not in macrophages. Functionally, both gp and human (h) IL-8 induced cell migration and ERK phosphorylation in HEK 293 cells expressing either receptor, whereas hGRO activated only cells expressing the clone 10 protein, confirming that clone 8 indeed coded for gpCXCR1. 125I-labeled hIL-8 bound to gpCXCR1 and addition of unlabeled hIL-8 completely abolished the binding; however, unlabeled gpIL-8 failed to compete against 125I-labeled hIL-8, strongly suggesting that the avidity of hIL-8 to gpCXCR1 is higher than that of gpIL-8. Identification and characterization of CXCR1 in the guinea pig will allow us to use this small animal model to evaluate the role of the IL-8/CXCR1 interactions and to examine the efficacy of CXCR1 antagonists in vivo. PMID- 16712934 TI - The genomic structure and a novel alternatively spliced form of porcine pTalpha chain. AB - A complete genomic nucleotide sequence for porcine pTalpha gene was obtained from a BAC clone, which revealed a novel exon 2 missing in human and murine counterparts. Cattle and dog genomic sequences showed the counterparts corresponding to porcine exon 2. Using thymocyte RNA and RT-PCR, three types of porcine pTalpha-chain cDNA sequences, pTalpha1, pTalpha2 and pTalpha3, were obtained. These three different cDNA sequences were alternatively spliced products with pTalpha1 consisting of exons 1, 2, 3, 4, and 5, pTalpha2 consisting of exons 1, 2, 4, and 5, and pTalpha3 consisting of exons 1, 2, 3 and the intron down stream of exon 3. pTalpha1 and pTalpha2 correspond to previously reported pTalphaa, and pTalphab, respectively, and pTalpha3 is reported for the first time. Using RT-PCR, pTalpha3 appeared expressed predominantly in the thymocyte RNA. The chromosome location of pTalpha was investigated using Radiation Hybrid Map and FISH, both of which revealed the location at SSC7q11-q12. PMID- 16712935 TI - Gene characterisation, isoforms and recombinant expression of carcinin, an antibacterial protein from the shore crab, Carcinus maenas. AB - Carcinin is a whey acidic protein (WAP) domain-containing antimicrobial protein produced by the circulating haemocytes of the shore crab, Carcinus maenas. Cloning of its full coding cDNA reveals that it shows some similarity to invertebrate defensins, has a valine-rich signal sequence followed by a defined cleavage site but no obvious acidic anionic 'pro' sequence. The C-terminus exhibits a unique cysteine array that is predicted to form six disulphide bonds in the tertiary structure. This 12 cysteine array arrangement is conserved in expressed sequence tags (ESTs) from related genera and seems to represent a novel tertiary structure amongst antimicrobial proteins (AMPs), unique to the Crustacea. There are at least five putative isoforms that arise through the transcription of a multi-exon gene. These isoforms do not arise as a result of alternate splicing of the exons, but by either the transcription of different alleles and/or single point mutation of the transcript at up to four loci in the gene. Several of the same transcripts have been found in different animals. The most commonly expressed transcript of the protein was recombinantly expressed in bacterial fusion system to a yield of ca. 2-3 microg ml(-1) of culture. In vitro expression with or without the leader sequence confirms the bioinformatic prediction that the stability of the mature protein is reduced when the leader sequence is removed. Carcinin is one of very few invertebrate AMPs characterised at the gene, transcript and protein level and to be recombinantly expressed in vitro in a bacterial system. PMID- 16712936 TI - From neurotoxic to chemosensory effects: new insights on acute solvent neurotoxicity exemplified by acute effects of 2-ethylhexanol. AB - Historically, acute solvent neurotoxicity was strongly related to reversible narcotic states that could be detected by neurobehavioral tests (e.g., simple reaction time). Nowadays, the occupational exposure to chemicals is markedly reduced and the avoidance of chemosensory effects is more important for the regulation of solvents. Exemplarily, this study examines if the chemosensory perception of 2-ethylhexanol is capable to distract performance in demanding neurobehavioral tasks. In two experiments three time-weighted average concentrations of 2-ethylhexanol (C(TWA): 1.5, 10, and 20 ppm) were investigated. In experiment A (n=24) variable concentrations over time (4h) were used, experiment B (n=22) investigated constant concentrations. The experiments were conducted in a 29 m3 exposure laboratory. Cross-over designs with randomized sequences of exposures were used. Among the 46 male participants 19 subjects reported enhanced chemical sensitivity; the other 27 subjects did not show this personality feature. During the exposure periods neurobehavioral tests were presented twice (beginning; end), the intensity of chemosensory perceptions were rated thrice. The intensity of chemosensory perceptions showed a clear dose dependency. Subjects' performance in the vigilance test was not affected by the different exposures. Moreover, the results of neurobehavioral tests measuring executive function were neither affected by the C(TWA) concentration nor by the exposure peaks. With increasing C(TWA), a subgroup of the chemically sensitive subjects showed deteriorated accuracy in a divided attention task. Especially the 20 ppm conditions were very annoying. Only during the constant 10 ppm condition the time courses of the annoyance and nasal irritation ratings indicated some adaptation. In general, with the applied neurobehavioral tests distractive effects of acute 2-ethylhexanol exposures up to 20 ppm could not be confirmed. In sensitive groups such distractive effects of irritating solvents might be conceivable and moreover, in cooperation with researchers from cognitive neuroscience more sensitive functions and tests should be developed and incorporated. PMID- 16712937 TI - Synthesis and characterization of palladium(II) and platinum(II) complexes with Schiff bases derivatives of 2-pyridincarboxyaldehyde. Study of their interaction with DNA. AB - Several Schiff bases ligand derivatives of 2-pyridincarboxyaldehyde and different amines, together with their palladium(II) and platinum(II) complexes have been synthesised and characterised. The aim of this study is to probe the influence of substituents beared on the pyridyl/toulene ring at different position to their possible antitumor activity. The amines used were o-, m-, p-toluidine and 4 hydroxyaniline. All the compounds were characterised by elemental analysis, FT-IR spectroscopy, 1H and 195Pt NMR spectroscopy and matrix assisted laser desorption/ionization time-of-flight mass spectroscopy. The formation of DNA adducts were analysed by circular dichroism and electrophoretic mobility. Atomic force microscopy images of the compounds with plasmid DNA pBR322 were also obtained. In all cases changes in the second and tertiary structure of DNA could be observed as a consequence of the covalent interaction of the palladium(II) or platinum(II) ions with the N of the nucleobases. However, there are not significant differences in the behavior of the complexes related to the position of the methyl groups or the presence of the OH group. Values of IC50 were also calculated for the platinum(II) complexes for several pairs of ovarian tumor cell lines which were either sensitive or resistant to cisplatin. Finally in vitro apoptosis studies for platinum(II) complexes with ovarian tumor cell lines A2780/A2780cisR were carried out. The results indicated interesting antiproliferative activity and significant apoptosis induction. PMID- 16712938 TI - Tetrathiolate coordination of germanium(IV) in a protein active site. AB - The tetracysteine metal coordination site of the rubredoxins from Clostridium pasteurianum (Cp) and Pyrococcus furiosus (Pf) are shown to stably bind the inorganic Ge(IV) ion. This is the first characterized coordination complex of tetravalent germanium with a biological macromolecule. Zn(II), Ga(III) and Ge(IV) substitution yields differential NMR chemical shifts for the 1H and 15N amide resonances throughout much of the protein structure. The differential shifts for the six backbone amides that hydrogen bond to the metal-coordinated sulfurs indicate that the pseudo 2-fold symmetry of the active site is more closely maintained in the hyperthermophile Pf rubredoxin than in its mesophile Cp homolog. These three metal substitutions form an isoelectronic series of small diamagnetic proteins for which reference structures are known to 1A resolution. These series provide a promising system to analyze theoretical predictions of the effects of differential charge distribution on chemical shifts from both proximal and long range interactions. PMID- 16712939 TI - Unusual mixed infection of thoracic empyema caused by Mycobacteria tuberculosis, nontuberculosis mycobacteria and Nocardia asteroides in a woman with systemic lupus erythematosus. AB - We report a 45-year-old woman of systemic lupus erythematosus (SLE) with thoracic empyema that was unusually infected by Mycobacterium tuberculosis (MTB), Nontuberculosis mycobacteria (NTM) concomitant with Nocardia asteroides. After a combined treatment of cotrimoxazole, clarithromycin and anti-tuberculosis drugs with a short-term of intravenous immunoglobulin (IVIG), the patient recovered from the critical illness. On the basis of the results in this case, we recommend a thorough survey of the probably concomitant infections of MTB and NTM in an immunocompromised patient with a known N. asteroid infection. In addition, an adjuvant intravenous immunoglobulin therapy may have beneficial effect in the control of infections in an SLE patient. PMID- 16712940 TI - Association of hypospadias and cardiac defect in a Parvovirus B19-infected stillborn: a causality relation? AB - Parvovirus B19 intrauterine infection is a known cause of hydrops fetalis and fetal death. It is also associated with congenital malformations, although the teratogenic potential seems to be low. Postmortem examination of a male stillborn of 29 gestational weeks revealed mild subcutaneous edema, malformed micropenis, perineoscrotal hypospadias and atrial septal defect, along with fetal erythroblastosis and villitis. Polymerase chain reaction detected Parvovirus B19 DNA genome in tissues from the fetus and the placenta, confirming the hypothesis of an intrauterine infection. PMID- 16712941 TI - Endothelial scavenger receptors. AB - In the past few decades, cDNAs for endothelial scavenger receptors that bind to negatively charged molecules, particularly acetylated low density lipoproteins (Ac-LDL), have been cloned by expression cloning using modified LDL as ligands. A prototypic members of endothelial scavenger receptor family, namely, scavenger receptor class B type I (SR-BI) has been characterized as a high density lipoprotein (HDL) receptor. Another prototypic member, CD36, has been determined as a multiple ligand receptor because it binds to oxidized LDLs (Ox-LDL), trombospondin, erythrocytes infected with Plasmodium falciparum, long-chain fatty acids, and Gram-negative and Gram-positive bacteria. Lectin-like oxidized LDL receptor-1 (LOX-1) has been discovered as the principal receptor that mediates the action of Ox-LDL in the vascular walls. Recently, the structure of oxidized phospholipids, originally found in Ox-LDL, and its molecular mechanism of action on endothelial cells were determined. Further, the use of genetically manipulated rodent models and the recent forward genetic screening technique revealed the physiological and pathological functions of these endothelial scavenger receptors in innate immunity and infection. In this review, the structure and function of these multiligand scavenger receptors of endothelial cells have been described mainly in relation with lipid metabolism. PMID- 16712942 TI - Determination of phylloquinone and cholecalciferol encapsulated in granulates formed by melt extrusion. AB - Vitamin K1 (phylloquinone) and vitamin D3 (cholecalciferol) play a dominant role in bone metabolism. Both vitamins are sensitive to ultraviolet radiation, oxygen and other environmental influences. For this reason a special extrusion technology was developed, that enables an encapsulation of these sensitive substances in a matrix of carbohydrates and hydrogenated carbohydrates. To exclude decomposition products possibly originating under process conditions quantitative analysis was carried out by HPLC/UV using a modified method based on United States Pharmacopoeia. Under the used chromatographic conditions it has to be possible to separate cis-phylloquinone, trans-phylloquinone and phylloquinone 2,3-oxide, as well as pre-cholecalciferol, cis-cholecalciferol and trans cholecalciferol. A silica column as stationary phase and a mixture of n-hexane and 1-amyl alcohol as mobile phase were used for quantification. UV detection ensued at 254 nm. A linear relationship between peak area and concentration was found over almost two orders of magnitude for cis-phylloquinone, trans phylloquinone and cholecalciferol. The detection limits (S/N 3) on column were 0.1 microg for phylloquinone and 0.4 microg for cholecalciferol. Analytical results showed that the vitamins were encapsulated sufficiently in the used carbohydrate matrix and that they were protected against environmental influences. After granulation process all of the samples tested met the pharmacopoeial requirements. PMID- 16712943 TI - Toward an integrative model of the spectrum of mood, behavioral and personality disorders based on fear and anger traits: II. Implications for neurobiology, genetics and psychopharmacological treatment. AB - Current psychiatry relies on a purely categorical paradigm for diagnosis of mental disorders that profoundly impacts research and clinical practice. However, high comorbidity rates and relative non-specificity of family history for psychiatric disorders suggests that this categorical approach fails to identify the underlying diathesis. As an attempt to overcome such limitations, we developed a bidimensional model based on fear and anger traits or temperaments which does not preclude the use of a categorical approach. As a result, it is hypothesized that mood, behavioral and personality disorders share a neurobiological substrate according to combinations of fear and anger traits. Both fear and anger, when excessive or deficient, lead to increased risk for mental disorders and should be considered in genetic, neurobiological and neuroimaging studies. Fear traits are much influenced by the amygdala and the serotonergic, noradrenergic and GABAergic systems, whereas anger seems to be mostly regulated by the nucleus accumbens and the dopaminergic and glutamatergic systems. Pharmacological treatments with antidepressants and anxiolytics can be considered as essentially restraint on fear, whereas lithium and alpha2 noradrenergic agonists would attenuate fear deficiency. Dopaminergic antidepressants and psychostimulants are anger enhancers and antipsychotics and mood stabilizers, such as divalproate and carbamazepine, may share antianger effects. Drugs effective for manic and depressive phases probably have both antianger and antifear effects. This framework may lead to a better understanding of the neurobiological basis of mental health and disease, providing an integrative approach for future research. PMID- 16712944 TI - Interpersonal psychotherapy for depression? The need to define its ecological niche. AB - BACKGROUND: Interpersonal psychotherapy (IPT) has long been viewed as an efficacious treatment for depression, with a large number of published studies allowing its efficacy and utility to be considered closely. METHODS: We review original studies and the one published meta-analysis of controlled efficacy trials, examining acute-phase and prophylactic studies, comparison studies with cognitive behaviour therapy, and studies evaluating its efficacy in combination with pharmacotherapy. RESULTS: We highlight difficulties in evaluating any psychotherapy, particularly when tested as having universal application for varying depressive conditions, but do identify circumstances where IPT may have specific salience. CONCLUSIONS: As for other treatments, IPT is unlikely to be a universal therapy for depression. This review offers suggestions as to where its preferential utility may lie, and argues for modified research paradigms to assist definition of its therapeutic niche. PMID- 16712945 TI - Do antidepressants t(h)reat(en) depressives? Toward a clinically judicious formulation of the antidepressant-suicidality FDA advisory in light of declining national suicide statistics from many countries. AB - Given that suicidality is a well-known symptom and outcome of untreated or inadequately treated depressive illness, the United States (US) Food and Drug Administration (FDA) warning of emergent suicidality in children and adolescents based on the antidepressant arm of placebo-controlled randomized trials (RCTs) has created understandable concern in clinical practice. The issues involved are of broader public health importance for all age groups. As in other branches of medicine, psychiatrists must always be vigilant of the rare risk of iatrogenesis when prescribing potent agents like antidepressants for patients with depressive disorders where the risk of suicidality is inherent. The overall evidence we review suggests that the widespread use of antidepressants in the new "SSRI-era" appear to have actually led to highly significant decline in suicide rates in most countries with traditionally high baseline suicide rates. The decline is particularly striking for women who, compared with men, seek more help for depression. Recent clinical data on large samples in the US too have revealed a protective effect of antidepressant against suicide. We argue that the discrepancy between RCTs (in children) and national and clinical suicide statistics (in adults) may reside in new provocative data documenting high rates of unrecognized pseudo-unipolar mixed states particularly in juvenile, but also in adult, clinical populations. Such an interpretation accords well with equally provocative data that bipolar II (which is often "mixed" in nature) may well represent a particularly vulnerable clinical substrate for suicidality. In this respect, the widespread (at least in the psychiatric sector) augmentation of antidepressants with benzodiazepines, atypical antipsychotics or mood stabilizers may represent one situation where current practice is superior to evidence-based medicine. We conclude that rather than being a threat, the judicious clinical use of antidepressants actually does serve to effectively treat and indeed protect depressed patients from suicidal outcome. The fact of being in treatment with regular clinical follow-up appears beneficial as well. PMID- 16712946 TI - Depression and help-seeking in a western sample of 'highly acculturated' Chinese and controls. AB - BACKGROUND: It is commonly reported that "the Chinese" have low rates of depression, partially reflecting a greater tendency to somatize and to be less likely to seek help. OBJECTIVE: To examine the impact of acculturation on depression reporting and help-seeking patterns. METHOD: We compared 50 highly acculturated Chinese with age and gender-matched non-Chinese control subjects in a western region on measures of state and lifetime depression, attributional interpretation of somatic cues and help-seeking. RESULTS: The highly acculturated Chinese did not differ in state depression rates or tendency to 'somatize'. There were trends for the Chinese to be less likely to view any depressive episode as a 'disorder' and to seek help for a psychological problem. CONCLUSION: Differences held to exist in the Chinese in acknowledging, reporting and seeking help for depression appear strongly culturally determined. PMID- 16712947 TI - Differences in characteristics between suicide victims who left notes or not. AB - BACKGROUND: Suicide notes (SN) are one of markers of the severity of a suicide attempt and are said to provide a valuable insight into the thinking of suicide victims before the fatal act [Shah, A., De, T., 1998. Suicide and the elderly. Int. J. Psychiat. Clin Pract. 2, 3-18]. To examine whether suicide victims who wrote notes (note writers: NW) differ from those who did not, we investigated the characteristics of a sample of more than 5000 Japanese suicides using multiple logistic regression analysis. METHODS: For all suicide victims (5161 cases), we examined the following information: gender, age, suicide method, reason for suicide, marital status, residential status, history of psychiatric disorders, previous suicidal behavior, physical disease, and content of suicide notes. RESULTS: Mean incidence of NW was 30.1% (male: 29.7%, female: 30.8%). NW in Japan had the following characteristics; higher proportion in female and living alone, suicide by more lethal methods such as carbon monoxide, hanging or sharp instruments. On the other hand, non-NW had tendencies to commit suicide for reasons of physical illness and psychiatric disorder, and/or history of previous psychiatric disorders. LIMITATIONS: This study is observational and discusses only completed, not attempted, suicide. Medical and psychiatric comorbidity are judged only by the history of diagnosis and the information about the problems in relationships is based not on valid criteria for inclusion. CONCLUSIONS: Although these findings show ethnic differences, it is possible that SN may be considered an indicator of a serious suicide attempt. Further studies of SN are needed to confirm this. PMID- 16712948 TI - The relationship between immunosuppressive activity and immunoregulatory cytokines in seminal plasma: influence of sperm autoimmunity and seminal leukocytes. AB - While the contributions of prostasomes, polyamines and prostaglandins to the immunosuppressive activity (ISA) of human seminal plasma have been well characterised, the contribution of immunoregulatory cytokines found in seminal plasma has received relatively little attention. Semen samples were collected from adult men displaying normospermic parameters, sperm antibodies or substantially elevated seminal leukocytes. Samples were processed through ultracentrifugation and dialysis (<3500Da) to remove prostasomes, polyamines and prostaglandins, and then assayed for ISA by an in vitro T lymphocyte inhibition assay, as well as by specific immunoassays for transforming growth factor beta(1) (TGFbeta(1)), interleukin-10 (IL-10), activin A and the activin-binding protein, follistatin. Seminal plasma from all groups retained substantial ISA following processing. Compared with normospermic men, this 'large' molecular weight ISA fraction was significantly increased in a subset of men with sperm antibodies, but was not altered in the group with elevated leukocytes. There was no relationship between ISA and any cytokine examined, and only TGFbeta(1) was present at levels sufficient to contribute to ISA. Inhibition with a TGFbeta specific antibody reduced ISA in seminal plasma by approximately 50%. Across all patients, TGFbeta(1) levels were positively correlated with sperm numbers in the ejaculate and with activin A, but not with follistatin or IL-10. Activin A and IL 10 also displayed a positive relationship, and elevated leukocytes was associated with a significant elevation of IL-10 and activin A, but not TGFbeta(1). It is concluded that 'large' molecular weight molecules, the most important of which appears to be TGFbeta(1), make a significant contribution to immunosuppression by human seminal plasma. PMID- 16712949 TI - Performance on the Wisconsin Card Sorting Test in schizophrenia and genes of dopaminergic inactivation (COMT, DAT, NET). AB - The aim of the study was to test an association between polymorphisms of genes connected with dopaminergic inactivation in prefrontal cortex [catechol-O methyltransferase (COMT), dopamine transporter (DAT), norepinephrine transporter (NET)], and performance on the Wisconsin Card Sorting Test (WCST), in schizophrenic patients. The number of perseverative errors (WCST-P), non perseverative errors (WCST-NP), completed corrected categories (WCST-CC), conceptual level responses (WCST-%CONC) and set to the first category (WCST-1st CAT) were measured. Genotyping was done for the Val108(158)Met polymorphism of the COMT gene (79 patients), the 3'UTR VNTR polymorphism of the DAT gene (124 patients) and the 1287 A/G polymorphism of the NET gene (63 patients). Male schizophrenic patients with Val/Val genotype of COMT obtained better results on WCST-P, while female patients had worse results on the WCST-NP compared with the remaining genotypes. There was a slight trend for patients with the A9/A9 genotype of DAT and with the A/A genotype of NET to perform better on some domains of the WCST, compared with other genotypes. A limitation to the interpretation of results could be small number of patients studied as well as variable psychopathological state and medication during cognitive testing. PMID- 16712950 TI - The Brief Motor Scale (BMS) for the assessment of motor soft signs in schizophrenic psychoses and other psychiatric disorders. AB - Subtle motor impairment is an important aspect of neurological soft signs (NSS) which are frequently found in psychiatric patients-particularly schizophrenic patients. On the basis of data collected in previous studies using two different NSS scales, the most reliable and discriminative signs of motor impairment were identified to construct a brief 10-item rating scale. Subsequently, the new scale was applied to a sample of subacute patients with schizophrenic psychoses (N = 82) and healthy controls (N = 33). Factor analysis identified two factors which were labeled "motor coordination" and "motor sequencing". Both the total score and the subscores showed high internal consistency and test-retest reliability. There were highly significant group differences on the total score and the subscores as well as on each individual task. The scale has high sensitivity (84.1%) and specificity (87.9%) indicating that it may provide an effective instrument for the screening of subjects with neurodysfunction as well as for monitoring motor dysfunction in the clinical course of psychiatric disorders. PMID- 16712951 TI - Relationship between parental bonding and mood disorder in six European countries. AB - The linkage between adverse parental child-rearing styles and the occurrence of mood disorders in adulthood has been investigated in a number of studies from different countries and cultural backgrounds. However, as direct cross-cultural comparisons hardly exist, little is known about cultural variations of this relationship. The European Study of the Epidemiology of Mental Disorders (ESEMeD) is a cross-sectional study in a stratified multi-stage random sample of 21,425 adults (18 years or older) from the general population of six European countries (Belgium, France, Germany, Italy, Netherlands, and Spain). Parental child-rearing styles were measured by means of a short form of the Parental Bonding Instrument (PBI) in 8,232 of the respondents. Based on the extraction and confirmation of a three-factor model of the PBI in a previous study, the association between parental bonding and mood disorders was studied by computing hierarchical nested multiple logistic regression models. The relationship between parental child rearing styles and mood disorders was mostly homogeneous across the six countries. The PBI dimensions maternal and paternal care had the strongest associations with mood disorders. A significant association of overprotection was observable only for the mother. There was no significant relationship between authoritarianism and the occurrence of mood disorders. In the European countries studied, the association between parental child-rearing styles and the occurrence of mood disorders appears not to be culture-dependent. It would be of interest to assess whether this also holds for the rest of Europe and non-European countries. PMID- 16712952 TI - Mental health service use among American Red Cross disaster workers responding to the September 11, 2001 U.S. terrorist attacks. AB - In this article, we explored 1) the extent of mental health (MH) service use by American Red Cross disaster relief workers, both before (lifetime) and 1 year after the September 11, 2001 terrorist attacks, and 2) demographic, disaster and MH variables predicting (1-year) post-September 11 MH service use in this population. A sample of 3015 Red Cross disaster workers was surveyed 1 year after the attacks, regarding demographic characteristics, MH service use before and since the attacks, and posttraumatic stress disorder (PTSD) symptoms. Findings revealed that while 13.5% used MH services before the attacks, 10.7% used services after. Variables increasing the likelihood of MH service use after the attacks included the following: no previous MH treatment, younger age, being divorced/widowed, and higher PTSD intrusion or hyperarousal symptoms. Findings support other recent research on MH service use after the September 11 attacks. PMID- 16712953 TI - Illicit drug use and anxiety disorders: findings from two community surveys. AB - The focus of this investigation was the relationship between anxiety disorders and lifetime use of amphetamines, cocaine, hallucinogens and heroin in two contemporaneous samples. Data from two independent community surveys conducted in the US (N=5877) and Ontario (N=8116) were used to assess whether a lifetime anxiety disorder diagnosis (social phobia, panic disorder, agoraphobia, specific phobia, and generalized anxiety disorder) was significantly associated with lifetime use of amphetamines, hallucinogens, cocaine, and heroin. Posttraumatic stress disorder was assessed only in the US survey. After controlling for sociodemographics, a significant association between any anxiety disorder diagnosis and lifetime stimulant use, cocaine use, and hallucinogen use was found in both surveys (OR approximately 1.5-3.0). Any anxiety disorder diagnosis was significantly associated with lifetime heroin use in the US survey (OR approximately 3.0). Clinicians and researchers need to be aware of the relationship between anxiety disorders and illicit drug use. PMID- 16712955 TI - Associations of phenylthiocarbamide tasting to alcohol problems and family history of alcoholism differ by gender. AB - Past research associating phenylthiocarbamide/propylthiouracil (PTC/PROP) taste status with alcoholism has produced equivocal results. Some have found higher proportions of nontasters among those with a family history of alcoholism than controls, whereas others have not. The purpose of this study was to investigate the relationship between PTC taste status, alcohol problems, and family history of alcoholism. A total of 244 undergraduate students participated in this study, with a gender distribution of 75% female and 25% male. We found support for our hypothesis that male supertasters would report fewer problems with alcohol and a less significant family history of alcoholism. Interestingly, we also found that female supertasters had a greater family history of alcoholism and more current problems associated with alcohol use. Implications for the genetic link between PTC taste status and alcoholism are discussed. PMID- 16712956 TI - Acute psychological stress alerts the adaptive immune response: stress-induced mobilization of effector T cells. AB - Influences of psychological stress on the acquired immune system have not consequently been investigated. We found acute psychological stress to cause an increase in CD56+ and CCR5+ effector T cells in the peripheral blood of healthy human subjects (N=22), while skin-homing CLA+ T cells decreased. At the same time, we observed a stress-induced decrease in CD45RA+/CCR7+ naive and CD45RA /CCR7+ central memory T cells, while CD45RA-/CCR7- effector memory and CD45RA+/CCR7- terminally differentiated T cells increased. This T cell redistribution translated into an increase in T cells expressing perforin/granzyme B and in Epstein-Barr virus-specific, cytomegalovirus-specific and influenza virus-specific CD8+ T cells. Thus, acute stress seems to promote the retention of less mature T cells within lymphoid tissue or skin while effector-type T cells are mobilized into the blood in order to be able to rapidly migrate into peripheral tissues. PMID- 16712954 TI - A cognitive neuroscience perspective on psychopathy: evidence for paralimbic system dysfunction. AB - Psychopathy is a complex personality disorder that includes interpersonal and affective traits such as glibness, lack of empathy, guilt or remorse, shallow affect, and irresponsibility, and behavioral characteristics such as impulsivity, poor behavioral control, and promiscuity. Much is known about the assessment of psychopathy; however, relatively little is understood about the relevant brain disturbances. The present review integrates data from studies of behavioral and cognitive changes associated with focal brain lesions or insults and results from psychophysiology, cognitive psychology and cognitive and affective neuroscience in health and psychopathy. The review illustrates that the brain regions implicated in psychopathy include the orbital frontal cortex, insula, anterior and posterior cingulate, amygdala, parahippocampal gyrus, and anterior superior temporal gyrus. The relevant functional neuroanatomy of psychopathy thus includes limbic and paralimbic structures that may be collectively termed 'the paralimbic system'. The paralimbic system dysfunction model of psychopathy is discussed as it relates to the extant literature on psychopathy. PMID- 16712957 TI - Analyses of a MS-associated haplotype encompassing the CCL3 gene. AB - Our previous studies showed the association of multiple sclerosis with the same marker haplotype encompassing the CCL3 gene in two independent sets of families. Here we present that sequencing of this haplotype and its flanking regions detected no new mutation, but 16 single nucleotide polymorphisms (SNP) and 1 insertion/deletion variant in both affected and unaffected individuals. Transmission distortion analyses of the newly identified variants in the second set of families revealed no individual marker association. In the absence of a single disease relevant variant within the MS associated haplotype and the surrounding linkage disequilibrium block, the highlighted haplotype may itself indicate a functionally relevant allelic combination or interaction. PMID- 16712958 TI - Myelin flow cytometry assay detects enhanced levels of antibodies to human whole myelin in a subpopulation of multiple sclerosis patients. AB - Antibodies directed against myelin components have been described in multiple sclerosis (MS). Accumulating evidence suggests that pathogenically relevant anti myelin antibodies bind conformational and post-translationally modified epitopes. However, the current methods to detect anti-myelin antibodies often do not allow recognition of such epitopes. We developed a flow cytometry-based assay to detect antibodies to whole human myelin (including conformational and post translationally modified epitopes). MS patients (n=152) showed enhanced serum levels of anti-myelin antibodies (total Ig, IgG and IgM) when compared to healthy donors (HD, n=40). Strikingly, approximately 50% of MS patients showed enhanced anti-myelin IgG levels. Anti-myelin IgG levels were not correlated with clinical parameters of disease. In the same population, serum antibody responses to recombinant myelin oligodendrocyte glycoprotein were comparable in MS patients and HD. PMID- 16712960 TI - (5R)-5-hydroxytriptolide (LLDT-8), a novel triptolide derivative, prevents experimental autoimmune encephalomyelitis via inhibiting T cell activation. AB - A novel triptolide derivative (5R)-5-hydroxytriptolide (LLDT-8) has been shown to have potent immunosuppressive activities. Here LLDT-8 was evaluated in experimental autoimmune encephalomyelitis (EAE), the model of multiple sclerosis (MS). LLDT-8 reduced the incidence and severity of EAE, which was associated with the inhibition of the MOG 35-55 lymphocyte recall response, anti-MOG 35-55 T cell responses, interleukin (IL)-2 and interferon (IFN)-gamma production. In vitro, LLDT-8 inhibited primary T cells proliferation, division, IL-2 and IFN-gamma production stimulated with anti-CD3/28. These findings highlight the fact that LLDT-8 prevents EAE by suppressing T cell proliferation and activation, with a potential for treatment of MS. PMID- 16712961 TI - Effects of GJB2 genotypes on the audiological phenotype: variability is present for all genotypes. AB - BACKGROUND AND AIM: Recent studies have revealed a genotype-phenotype correlation for mutations in the GJB2 gene. Since ethnic difference may have an effect for the degree of hearing loss due to background genes, we aimed to search for confirmation of previously suggested genotype-phenotype correlation in GJB2 deafness in the Turkish population. METHODS: Pure tone audiograms of 63 unrelated probands with GJB2-associated hearing loss having 15 different mutations were obtained and evaluated for correlation between the degree of hearing loss and genotypes. RESULTS: Three GJB2 genotypes identified in more than one family were homozygous c.35delG (44 probands), homozygous p.E120del (four probands) and c.[35delG]+[IVS1+1G>A] (two probands). No statistical difference for the degree of hearing loss was observed when the genotypes were compared individually or grouped according to their effects on the protein. The most likely explanation for this result is the relatively small size of the studied population. Degree of hearing loss was variable in c.35delG and p.E120del homozygotes. Intra-familial phenotypic variability was present for some genotypes. The detailed audiological data for homozygous p.E120del and c.[35delG]+[328delG] genotypes are reported for the first time in this study. CONCLUSION: Previously reported genotype-phenotype correlations for the GJB2 deafness should be cautiously interpreted during the clinical counseling since variability in the degree of hearing loss is present for all GJB2 genotypes. PMID- 16712962 TI - Development of real-time and conventional RT-PCR assays for the detection of potato yellow vein virus (PYVV). AB - Potato yellow vein virus (PYVV) is considered a quarantine pathogen in the European and Mediterranean Plant Protection Organization (EPPO) area. This virus is widespread and damaging at its centre of origin in South America. Current detection methods are either time-consuming or difficult to interpret. This paper reports the development of a sensitive, high throughput, real-time reverse transcription (RT)-PCR assay, based on TaqMan chemistry, suitable for PYVV detection. In addition, a reliable conventional RT-PCR assay for PYVV detection is also presented. Although less sensitive (1000 times less sensitive in direct comparison), this method requires less sophisticated equipment and as such should be a useful alternative to the real-time technique in some testing laboratories. The two assays presented here could assist in the implementation of quarantine measures for PYVV identification and in routine indexing of PYVV for the production of virus-free seed potatoes in areas of South America where the virus is highly damaging. PMID- 16712963 TI - Bovine herpesvirus 4 based vector interaction with liver cells in vitro and in vivo. AB - Gene transfer into hepatocytes is highly desirable for the long-term goal of replacing deficient proteins and correcting metabolic disorders. Bovine herpesvirus 4 (BoHV-4) based vector capability to transduce rat liver cells in vitro and in vivo was assessed. For the in vitro study, a buffalo rat liver cell line was successfully transduced by BoHV-4 and although did not show toxicity, the immediate early two viral gene was transcribed and cells harboring the intact viral genome could be pharmacologically selected, but no viral replication took place. For the in vivo study, adult male rats were inoculated intraportally and intraparenchimally with a BoHV-4 expressing enhanced green fluorescent protein and liver sections were analyzed through fluorescent microscopy. Although the liver parenchyma could not be transduced, the endothelial layer of the liver vasculature showed a robust transgene expression without toxicity. Successful BoHV-4 based vector transduction of primary cultures of rat hepatocytes suggests that extrinsic factors, and not hepatocytes per se, are the cause of such lack of transducibility. The present study serves as a starting point for study of the use of BoHV-4 based vectors to target gene delivery to vascular endothelial cells. PMID- 16712964 TI - A comparison of enhanced green fluorescent protein expression induced by immediate-early cytomegalovirus (IE-CMV) and gG pseudorabies virus (gG-PRV) promoters, using pseudorabies virus amplicons as vectors. AB - This study compares the expression efficiencies of the IE-CMV and gG-PRV promoters following their transfection into cultured human and monkey cells, using pseudorabies virus amplicons as vectors and enhanced green fluorescence protein (EGFP) as an expression marker. EGFP expression was similarly strong with both promoters. Pseudorabies virus amplicons appear to be useful vectors in gene expression studies due to their replication in the presence of helpers and their wide range of cellular hosts. PMID- 16712966 TI - Aryl furano pyrimidines: the most potent and selective anti-VZV agents reported to date. AB - Bicyclic aryl furano pyrimidines represent the most potent anti-VZV agents reported to date. Lead compounds have EC50 values in vitro as low as 0.1 nM and selectivity index values exceeding one million. They have an absolute requirement for VZV thymidine kinase (TK) and most likely act as their phosphate forms. Some structural modification, such as aryl substitution, is tolerated, while little sugar modification is acceptable. We herein summarise their biological profiles and structure activity relationships as discovered to date. PMID- 16712965 TI - Looking back: corticothalamic feedback and early visual processing. AB - Although once regarded as a simple sensory relay on the way to the cortex, it is increasingly apparent that the thalamus has a role in the ongoing moment-by moment processing of sensory input and in cognition. This involves extensive corticofugal feedback connections and the interplay of these with the local thalamic circuitry and the other converging inputs. Here, using the feline visual system as the primary model, some of the latest developments in this field are reviewed and placed in the perspective of an integrated view of system function. Cortical feedback mediated by ionotropic and metabotropic glutamate receptors, and effects mediated by the neuromodulator nitric oxide, all have a role in integrating the thalamic mechanism into the cortical circuit. The essential point is that the perspective of higher-level sensory mechanisms shifts and modulates the thalamic circuitry in ways that optimize abstraction of a meaningful representation of the external world. This review is part of the TINS special issue on The Neural Substrates of Cognition. PMID- 16712967 TI - Cell line dependency for antiviral activity and in vivo efficacy of N methanocarbathymidine against orthopoxvirus infections in mice. AB - A novel carbocyclic thymidine analog, N-methanocarbathymidine [(N)-MCT], was evaluated for inhibition of orthopoxvirus infections. Efficacy in vitro was assessed by plaque reduction assays against wild-type and cidofovir-resistant strains of cowpox and vaccinia viruses in nine different cell lines. Minimal differences were seen in antiviral activity against wild-type and cidofovir resistant viruses. (N)-MCT's efficacy was affected by the cell line used for assay, with 50% poxvirus-inhibitory concentrations in cells as follows: mouse=0.6 2.2 microM, rabbit=52-90 microM, monkey=87 to >1000 microM, and human=39-220 microM. Limited studies performed with carbocyclic thymidine indicated a similar cell line dependency for antiviral activity. (N)-MCT did not inhibit actively dividing uninfected cells at 1000 microM. The potency of (N)-MCT against an S variant thymidine kinase-deficient vaccinia virus was similar to that seen against S-variant and wild-type viruses in mouse, monkey, and human cells, implicating a cellular enzyme in the phosphorylation of the compound. Mice were intranasally infected with cowpox and vaccinia viruses followed 24h later by intraperitoneal treatment with (N)-MCT (twice a day for 7 days) or cidofovir (once a day for 2 days). (N)-MCT treatment at 100 and 30 mg/kg/day resulted in 90 and 20% survival from cowpox virus infection, respectively, compared to 0% survival in the placebo group. Statistically significant reductions in lung virus titers on day 5 occurred in 10, 30, and 100mg/kg/day treated mice. These same doses were also active against a lethal vaccinia virus (WR strain) challenge, and protection was seen down to 10mg/kg/day against a lethal vaccinia virus (IHD strain) infection. Cidofovir (100mg/kg/day) protected animals from death in all three infections. PMID- 16712968 TI - Induction and expression of salt appetite: effects on Fos expression in nucleus accumbens. AB - Sodium depletion is a strong natural motivator that creates a pronounced sodium appetite and has been shown to activate neural regions associated with fluid and sodium balance. However, it is not known whether sodium appetite affects the mesolimbic circuitry associated with reward motivation. The present studies examined expression of the immediate early gene Fos in the nucleus accumbens (NAc) as a marker of neuronal activation following the induction and expression of furosemide-induced sodium appetite. During sodium appetite expression, sham drinking and normal drinking were used to dissociate effects of NaCl taste stimulation from the repletion that follows absorption of sodium. These studies revealed that the combination of NaCl taste stimulation and persistent sodium depletion experienced by sham-drinking animals dramatically activates the NAc, while neither induction nor expression of sodium appetite alone is sufficient to increase Fos expression in this region. Results are discussed in terms of current theories of reward motivation. PMID- 16712969 TI - The effects of chronic administration of quetiapine on the methamphetamine induced recognition memory impairment and dopaminergic terminal deficit in rats. AB - Previous studies have suggested that quetiapine, a new atypical antipsychotic drug, may have beneficial effects on cognitive impairment and be a neuroprotectant in treating neurodegenerative diseases. In the present study, we investigated the therapeutic effects of chronic administration of quetiapine on methamphetamine (METH)-induced recognition memory impairment and dopaminergic terminal neurotoxicity in rats. Rats were pretreated with METH (5 mg/kg; s.c.) four times at 2-h intervals while their body temperature was monitored. Fifteen minutes after the last METH injection, rats were administered quetiapine (10 mg/kg/day; i.p.) for 28 days. One day after the last quetiapine injection, rats were trained and tested on an object recognition task on days 29 and 30. Finally, on day 31, rats were sacrificed for immunohistochemistry, 1 day after the object recognition task. METH induced hyperthermia, recognition memory impairment and a decrease of tyrosine hydroxylase immunoreactivity in the caudate putamen (CPu) of striatum. Quetiapine attenuated the METH-induced hyperthermia. Furthermore, chronic post-treatment of quetiapine reversed the METH-induced memory impairment and dopaminergic terminal deficit. These findings suggest that quetiapine may have therapeutic effects in the treatment of cognitive impairment and neurodegeneration induced by METH. PMID- 16712970 TI - Exploration in a dark open field: a shift from directional to positional progression and a proposed model of acquiring spatial information. AB - Exploration in a dark open field undergoes three progressive changes: (i) an initial phase of spending equal amounts of time in various zones of the arena changes to staying in the corners, and ultimately spending most of the time in one corner; (ii) travel paths are first circular and scattered all over the arena, but gradually become anchored to one corner at which they start and end; (iii) traveled distance gradually decreases to that of the initial level seen in a lit open field. Altogether, rodents shift from a 'looping' exploration mechanism with feeble coupling with the environment, to 'home base' exploration which is firmly anchored to the environment. This shift also involves switching from momentary and sporadic to repeated returns to a specific, presumably familiar place, to which the animal navigates back from various other places. We suggest that this switching illustrates navigation first by directional and then by positional environmental cues, as hypothesized in the 'parallel map theory'. We also suggest that the transition from looping to home base behavior is part of a hierarchal construction of space representation via three modes of spatial information processing: (i) piloting--sequential processing, based on moving from one landmark to the next; (ii) orienting--parallel processing, based on moving from one point to the next, with the same starting and ending point; (iii) navigating--continuous processing, based on continuously updating the position in relation to several locations in the environment (map navigation). PMID- 16712971 TI - Defensive responses of Wistar and Sprague-Dawley rats to cat odour and TMT. AB - Cat odour and trimethylthiazoline (TMT) are two predator odours commonly used to study defensive behaviour in rats. However their reported efficacy varies markedly across laboratories. We assessed whether rat strain differences might explain such variation. Wistar and Sprague-Dawley rats were tested for unconditioned and conditioned responses to both odours. Cat odour produced robust unconditioned and conditioned defensive behaviour, with notably stronger effects in Wistar rats. TMT produced limited unconditioned avoidance, but failed to elicit conditioned responses in either strain. Results support suggestions that faeces-derived odours such as TMT are less predictive of a predator threat than those derived from fur or skin, and identify the possibility that strain differences affect the defensive response seen to predator odours. PMID- 16712972 TI - Behavioral characterization of CD26 deficient mice in animal tests of anxiety and antidepressant-like activity. AB - CD26 exhibits a dipeptidylpeptidase-IV function (DPPIV) which regulates neuropeptide activity by N-terminal processing. Because abnormal plasma DPPIV was associated in mammals with behavioral changes, we examined the behavior of CD26-/ mice resulting from targeted inactivation of the gene. These animals had a decreased immobility in the forced swim and tail suspension tests, indicating a reduced depression-like behavior. We addressed some factors that could affect these results. No major differences between mutants and controls were observed in the black/white box test that investigates anxiety. In the hole-board apparatus that explores both curiosity and anxiety, CD26-/- mice of both genders made significantly more head dips than controls. In a motor activity test, mutants displayed higher horizontal and vertical activities i.e. increased novelty induced behavioral activation. We conclude that DPPIV inactivation in mice broadly leads to an antidepressant-like and hyperactive phenotype. PMID- 16712973 TI - Estrogen reduces the severity of autonomic dysfunction in spinal cord-injured male mice. AB - Autonomic dysreflexia is an autonomic behavioural condition that manifests after spinal cord injury (SCI) and is characterized by acute, episodic hypertension following afferent stimulation below the level of the injury. Common triggers of autonomic dysreflexia include colorectal distension (CRD), and various somatic stimuli. The development of autonomic dysreflexia is dependent, in part, upon the degree of intraspinal inflammation and the resultant spinal neuroplastic changes that occur following SCI. 17beta-estradiol (E) has neuroprotective, anti inflammatory and smooth muscle relaxant properties, and is therefore a candidate drug for the treatment and/or prevention of autonomic dysreflexia. Autonomic dysreflexia was assessed in adult male mice treated with E. We investigated whether E could be acting centrally by altering: (1) the size of the small diameter primary afferent arbor, (2) the degree of microglia/macrophage infiltration at the site of the injury, or (3) the amount of fibrous scarring present at the injury site. To determine whether E could be working through uncoupling protein-2 (UCP-2), a protein involved with inflammation and regulated by estrogen in some tissues, autonomic dysreflexia was assessed in E-treated adult male mice lacking UCP-2 (UCP-2 KO). 17beta-estradiol was equipotent at reducing autonomic dysreflexia in both UCP-2 KO and WT mice following CRD but not tail pinch. We have shown that E reduces autonomic dysreflexic responses to visceral but not somatic stimulation in male mice independent of the size of the primary afferent arbour, the degree of chronic inflammation, and the presence of UCP-2. PMID- 16712974 TI - Differential alcohol drinking behaviour and dependence in the Naples low- and high-excitability rat lines. AB - Alcoholism is composed of a series of phenomena, which can be approached in model systems. Our aim was to investigate psychogenetics of alcohol drinking behaviour and dependence in the Naples rat lines. Thus, adult male rats of the Naples low- (NLE), high-excitability (NHE) and random-bred control (NRB) lines had 24-h access to a four-bottle system with tap water and 5, 10 and 20% alcohol solutions. Total voluntary alcohol intake and preference were measured during a 6 week period. In order to analyze behavioural dependence, two abstinence periods were introduced, followed by re-exposure to the bottles. The first was a pure re test and the second was a quinine test (adding 0.02% quinine alcohol solution). Finally, to verify the role of opioids on alcohol drinking behaviour, rats received naltrexone (5 mg/kg i.p.). Thirty minutes later, they had access for 2 h to a three-bottle system with water, 5 and 20% alcohol solution, respectively. Results indicate that (i) total alcohol intake on day 1 was higher in both NLE/NHE, becoming higher in NLE rats during the 6-week period, (ii) different alcohol preference patterns in the Naples lines were observed over time of testing, (iii) during re-test, NLE rats showed neither reversibility nor alcohol deprivation effect, (iv) during quinine test, only NLE rats reduced alcohol intake to a lower extent and (v) naltrexone reduced by 50% the intake of the preferred solution. Therefore, the NLE rats appear as a new model for studying the neural substrates of the dependence behaviour. PMID- 16712975 TI - Acute perinatal asphyxia at birth has long-term effects on behavioural arousal and maternal behaviour in lactating rats. AB - This study analysed the long-term consequences of an asphyctic event at birth on maternal behaviour and emotionality in rats. Pregnant Wistar rats were delivered by Caesarean section and the pups, still in the uterus horns, were placed into a water bath at 37 degrees C for periods of 0 (Caesarean delivery, CD) or 20 min (asphyxia, CD+20). Control subjects were born by vaginal delivery (VD). Subsequently, pups were given to surrogate mothers, weaned at 21 days, and mated at adulthood. After giving birth, differences in maternal competence and behavioural arousal were assessed observing: (i) maternal behaviour after reunion with the pups following a 3h separation, on postpartum day (PPD) 4; (ii) behaviour in an elevated plus-maze test, on postpartum day 7; and (iii) performance in a fear conditioning test, in which subjects learned to associate a conditional stimulus with an aversive unconditioned stimulus (postpartum days 7 8). Results indicate that subjects in the CD+20 group showed a deficit in maternal care, taking a longer time to retrieve the whole litter and often failing to retrieve all pups. Both CD and CD+20 groups showed higher behavioural activity in the plus-maze. In addition, when tested in the fear conditioning paradigm, the CD+20 group showed a lower latency to perform freezing behaviour in the auditory cue trial. The changes in behavioural arousal described suggest that the dopaminergic system may be a potential neurochemical target for an early hypoxic insult and indicate maternal behaviour as a useful endpoint to study the effects of early birth insult on brain function. PMID- 16712976 TI - The effect of hypertension on serum nitric oxide and vascular endothelial growth factor concentrations. A study in DOCA-Salt hypertensive ovariectomized rats. AB - CardioVascular Disease (CVD) accounts for considerable mortality and morbidity in developed countries. Most of the common forms of CVD, such as hypertension, are caused by functional and structural changes in endothelial function. This study was designed to study the effect of hypertension on serum Nitric Oxide (NO) and Vascular Endothelial Growth Factor (VEGF) concentrations in DOCA-Salt hypertensive ovariectomized rats. Thirty female rats were ovariectomized. Blood samples were taken and the animals were divided into hypertensive and control groups. Hypertension was induced by DOCA-Salt method. DOCA was injected 30 mg/kg of body weight subcutaneously, twice a week with NaCl 1% instead of tap water for drinking throughout the experiment. The control group received normal saline injection with usual drinking water. Results showed that serum NO concentration in DOCA-Salt hypertensive rats was lower than the control group (18.35 +/- 5.31, 45.01 +/- 12.54 micromol/l, respectively) (p < 0.05). Also, the mean serum VEGF concentration was raised after induced hypertension (120.55 +/- 8.11 vs. 88.58 +/ 2.24 pg/ml) (p < 0.05). In conclusion, reduced serum NO and increased serum VEGF concentrations in hypertensive animals support the concept of endothelial dysfunction in hypertensive subjects. PMID- 16712977 TI - Vagally mediated cholestatic and choleretic effects of centrally applied Endothelin-1 through ETA receptors. AB - The role of Endothelin-1 (ET-1) in the central nervous system is not fully understood yet although several studies strongly support its neuromodulatory role. A high density of endothelin receptors is present in the dorsal vagal complex that is the major site for the regulation of the digestive function. Therefore in the present study we sought to establish the role of ET-1 in the central regulation of bile secretion in the rat. Intracerebroventricular ET-1 injection exhibited opposite behaviors on spontaneous bile secretion according to the dose administered. Lower doses of ET-1 (1 fM) increased bile flow and bicarbonate excretion whereas higher doses (1 nM) decreased bile flow and bile acid output. Both the choleretic and the cholestatic effects of ET-1 were abolished in animals pretreated with icv BQ-610 (selective ETA antagonist) but not with BQ-788 (selective ETB antagonist). In addition, truncal vagotomy but not adrenergic blockade abolished ET-1 effects on bile secretion. Brain nitric oxide was not involved in ET-1 response since L-NAME pretreatment failed to affect ET-1 actions on the liver. Portal venous pressure was increased by centrally administered ET-1 being the magnitude of the increase similar with low and high doses of the peptide. These results show that centrally applied ET-1 modified different bile flow fractions independent of hemodynamic changes. Lower doses of ET-1 increased bile acid independent flow whereas higher doses decreased bile acid dependent flow. Vagal pathways through the activation of apparently distinct ETA receptors mediated the cholestatic as well as the choleretic effects induced by ET-1. Present findings show that ET-1 participates in the central regulation of bile secretion in the rat and give further insights into the complexity of brain-liver interaction. PMID- 16712979 TI - Role of NPR-C natriuretic receptor in nitric oxide system activation induced by atrial natriuretic peptide. AB - Atrial natriuretic peptide (ANP) exerts its hypotensive, natriuretic and diuretic effects, almost in part, through the activation of nitric oxide synthase (NOS). The aim was to investigate the natriuretic receptor type and the signaling cascade involved in NOS activation induced by ANP. Male Wistar rats were sacrificed and NOS activity was determined in kidney, aorta and heart with L [U14C]-arginine, as substrate. ANP and cANP (4-23), a selective NPR-C ligand, increased NOS activity in all tissues. ANP induced a more marked activation in aorta and kidney than cANP (4-23), but no difference in atria NOS activation was observed. NOS activity induced by both peptides was blunted by nifedipine (L-type channel blocker) and calmidazolium (calmodulin antagonist) in heart and aorta. In kidney, nifedipine and calmidazolium abolished NOS activity stimulated by cANP (4 23) but only partially inhibited NOS activity elicited by ANP. Gi inhibition with pertussis toxin abolished NOS activity stimulated by ANP and cANP in atria but only partially inhibited the increased NOS activity induced by ANP and cANP in kidney, aorta and ventricle. Our results show that NPR-C receptor would mediate the activation of NOS by ANP in atria. In kidney, aorta and ventricle, NOS activation would also involve NPR-A and/or B. ANP would interact with NPR-C coupled via Gi to activation Ca2+ -dependent NOS. PMID- 16712978 TI - Role of kappa- and delta-opioid receptors in the antinociceptive effect of oxytocin in formalin-induced pain response in mice. AB - Oxytocin has been implicated in the modulation of somatosensory transmission such as nociception and pain. The present study investigates the effect of oxytocin on formalin-induced pain response, a model of tonic continuous pain. The animals were injected with 0.1 ml of 1% formalin in the right hindpaw and the left hindpaw was injected with an equal volume of normal saline. The time spent by the animals licking or biting the injected paw during 0-5 min (early phase) and 20-25 min (late phase) was recorded separately. Oxytocin (25, 50, 100 microg/kg, i.p.) dose dependently decreased the licking/biting response, both in the early as well as the late phases. The antinociceptive effect of oxytocin (100 microg/kg, i.p.) was significantly attenuated in both the phases by a higher dose of the non selective opioid receptor antagonist naloxone (5 mg/kg, i.p.), MR 2266 (0.1 mg/kg, i.p.), a selective kappa-opioid receptor antagonist and naltrindole (0.5 mg/kg, i.p.), a selective delta-opioid receptor antagonist but not by a lower dose of naloxone (1 mg/kg, i.p.) or beta-funaltrexamine (2.5 microg/mouse, i.c.v.), a selective mu-opioid receptor antagonist. Nimodipine, a calcium channel blocker (1 and 5 mg/kg, i.p.) produced a dose-dependent analgesic effect. The antinociceptive effect of oxytocin was significantly enhanced by the lower dose of nimodipine (1 mg/kg, i.p.) in both the phases. Chronic treatment with oxytocin (100 microg/kg/day, i.p. daily for 7 days) did not produce tolerance in both the phases of formalin-induced pain response. The results thus indicate that oxytocin displays an important analgesic response in formalin test; both kappa- and delta opioid receptors as well as voltage-gated calcium channels seem to be involved in the oxytocin-induced antinociception. PMID- 16712980 TI - The evaluation of depression in the elderly: a modification of the Geriatric Depression Scale (GDS). AB - A modified version of the GDS was created to manage the difficulties often encountered with very old respondents. The scale was applied to 195 old persons (mean age of 78 years), males and females, community-dwelling, and nursing home residents. Results indicate that the participants comprehend significantly better the modified than the original version of the scale, and both versions retain comparably good psychometric properties. PMID- 16712981 TI - Effectiveness of validation therapy (VT) in group: preliminary results. AB - VT is one of the best known psychosocial treatment for elderly affected by dementia. Notwithstanding its wide use, its efficacy is still a controversial issue, especially in comparison with other approaches. This study of 30 elderly subjects in a nursing home compared the effects of VT, sensorial reminiscence (SR) and no treatment on cognitive, functional, and affective status of the participants. The results indicated an improvement in the global functioning of the two treatment groups, compared to the control group, but the differences did not reach statistical significance. Significant within-group effects could be observed, where the SR treatment was the most effective in improving cognitive, affective, and behavioral status; the VT-group showed a reduction of the behavioral disturbances; the participants at the control group demonstrated a slight deterioration at all the three levels. Considerations are made about the implication of VT for the caregivers, as possible mediator of its effect on elderly. PMID- 16712982 TI - Plaque ruptures in stable angina pectoris compared with acute coronary syndrome. AB - BACKGROUND: Plaque rupture is more frequently observed in patients with acute coronary syndrome (ACS) rather than in patients with stable angina pectoris (SAP). Consequently, studies regarding plaque rupture, which occurred in SAP patients, are rare. Therefore, we evaluated the frequency and axial location of plaque ruptures in SAP patients and compared them with those in ACS patients. METHODS: Three hundred ninety-two patients (231 ACS and 161 SAP patients) who were scheduled for coronary intervention underwent three-vessel intravascular ultrasound (IVUS) study. IVUS criteria for plaque rupture were a plaque contained a cavity that communicated with the lumen with an overlying residual fibrous cap fragment. Using motorized IVUS transducer pullback in all three coronary arteries, the distance between each coronary plaque rupture segment and the respective coronary ostium was measured. RESULTS: Plaque ruptures were detected in 206 of 392 patients who underwent three-vessel intravascular ultrasound examination. At least one plaque rupture in any coronary artery was noted in 48 (30%) SAP and 158 (68%) ACS patients (p<0.001). In both ACS and SAP patients, plaque ruptures were clustered mainly in the proximal segments of the left anterior descending artery and in the proximal and distal segments of the right coronary artery. CONCLUSIONS: At least one plaque rupture in any coronary artery was noted in 30% of SAP patients. Like in ACS patients, plaque ruptures were clustered mainly in the proximal segments of the left anterior descending artery and in the proximal and distal segments of the right coronary artery in SAP patients. PMID- 16712983 TI - Protective effects of pyrrolidine dithiocarbamate on myocardium apoptosis induced by adriamycin in rats. AB - BACKGROUND: Effects of pyrrolidine dithiocarbamate (PDTC) on programmed cell death are controversial. It is unclear if PDTC has the protective effects on myocardial apoptosis induced by adriamycin (ADR) in rats. The present study was undertaken to study the protective effects of pyrrolidine dithiocarbamate (PDTC) on myocardium apoptosis induced by adriamycin (ADR) in rats and its mechanisms. METHODS: Forty male Wistar rats were randomly divided into five groups: control, ADR, ADR+PDTC 50 mg/kg, ADR+PDTC 100 mg/kg and ADR+PDTC 200 mg/kg group. Myocardial apoptosis was detected by electron microscopic examination and TUNEL assay. Myocardium p53 gene expression was examined by RT-PCR analysis. Location and distribution of p53 was observed by immunohistochemical assay. Myocardial expression of p53 protein was assessed by Western blot analysis. Activity of NF kappaB was evaluated by Electrophoretic Mobility Shift Assay. RESULTS: Myocardial apoptotic index, expression of p53 mRNA, expression of p53 protein and the binding activity of NF-kappaB decreased significantly in ADR+PDTC groups compared with ADR group. All these change were significantly correlated with dose of PDTC. CONCLUSION: PDTC has preventive effects on myocardial apoptosis induced by ADR, which is probably associated with inhibiting binding activity of NF-kappa B and further regulating apoptosis-related gene expression and translation, and inhibiting myocardial apoptosis. PMID- 16712984 TI - Early discharge after acute myocardial infarction in the current clinical practice. Community data from the AMI-Florence Registry, Italy. AB - BACKGROUND: Clinical trials suggested feasibility and safety of early discharge after ST-segment elevation acute myocardial infarction (STEMI) for selected patients. Current United States and European guidelines recommend early discharge for uncomplicated AMI. The present study was aimed to assess early discharge in the current clinical practice for STEMI patients. METHODS: Patients enrolled in the AMI-Florence Registry (Italy), a prospective, observational, population-based study performed in the early 2000s, were analysed. The proportion of STEMI patients eligible for early discharge and of those actually discharged early, patient features influencing early discharge and outcomes (mortality, reinfarction or urgent revascularization during the first year of follow-up) were assessed. RESULTS: Of 751 STEMI patients, 59% were classified as eligible for early discharge, according to previously established criteria. Among patients considered eligible, those actually discharged early dropped from 33.1% in age <70 years to 15.9% in age 70-79 years and to 11.7% in age 80+ years. Of eligible patients, 26% were actually discharged within 4 days. Age 70+ years (reference: 69 years or younger; 70-79 years: OR: 0.40, 95% CI: 0.22-0.73; 80+ years: OR: 0.33, 95% CI: 0.14-0.76) and diabetes (OR: 0.48, 95% CI: 0.24-0.98) were negative independent predictors; and coronary reperfusion (OR: 2.92, 95% CI: 1.54-5.53) or firstly admitted to teaching hospital (OR: 1.68, 95% CI: 1.03-2.74) were positive predictors, of the multivariate probability of being early discharged. Among patients eligible for early discharge, 1-year and 3-year mortality did not differ significantly between patients actually, and not, early discharged. After 1 year of follow-up, no deaths were observed among patients early discharged. CONCLUSION: This study confirms, in a setting of current clinical practice, the reliability of criteria for identifying patients eligible for early discharge. Besides, the results confirmed the safety of this practice in selected patients. About 1/4 of eligible patients are actually early discharged, confirming the existence of important opportunities to improve the efficiency in STEMI management. PMID- 16712985 TI - A quantitative assessment of the risk of exposure to bovine spongiform encephalopathy via meat-and-bone meal in Japan. AB - The feeding of meat-and-bone meal (MBM) derived from cattle infected with bovine spongiform encephalopathy (BSE) is a major source of BSE infection. The risks of BSE infection via MBM in Japan were examined quantitatively to estimate infectivity to cattle via MBM derived from a single clinically infected animal being rendered. Three routes of exposure were modeled: (i) feeding cattle concentrates containing MBM as an ingredient, (ii) feeding cattle concentrates contaminated with MBM from non-ruminant feed at feed plants and (iii) directly feeding MBM in supplemental form to cattle on farms. The effectiveness of measures designed to restrict the feeding of ruminants with ruminant MBM (feed restriction) as well as differences in the risk of exposure among regions were examined using the model. The model revealed that the median total infectivity fed to dairy cattle via MBM derived from one infected animal was approximately 0.49 cattle oral ID(50) (5th percentile=0.43ID(50), 95th percentile=0.54ID(50)). This value was reduced by 55% after the addition of MBM to cattle concentrates was restricted in 1996. The risk of exposure in dairy cattle was twice that in beef cattle. Comparisons of regional differences in exposure risk indicated that the risk was highest in a region where 14 of the 20 BSE cases reported to date were born. Our model suggested that the routes of exposure via MBM were unlikely to result in increased propagation of BSE in Japan. Furthermore, despite some regional variation, the risk of exposure declined further after the feed restriction was imposed in 1996. PMID- 16712986 TI - Optimal risk adjustment with adverse selection and spatial competition. AB - Paying insurers risk-adjusted prices for covering different individuals can correct selection incentives and induce the market to provide optimal insurance policies. To calculate the optimal risk-adjusted prices we need to know (a) what the optimal policies are; (b) how much they cost; and (c) how competitive the market is. We examine these issues in a model with spatial heterogeneity and adverse selection. Market equilibrium is characterized, and delivery of the socially optimal insurance policies is possible, as long as providers are paid risk-adjusted fees for each individual they serve. When the payment can be made on the basis of an individual's risk, it should be sufficient to cover the expected cost of the socially optimal policy for that person, plus a mark-up. If payments can be made only on the basis of a partially informative signal, the optimal risk-based payments should be adjusted according to a simple linear transformation, identified by Glazer and McGuire [Glazer, J., McGuire, T., 2000. Optimal risk adjustment of health insurance premiums: an application to managed care. PMID- 16712987 TI - Insurer-provider integration, credible commitment, and managed-care backlash. AB - People distrust managed care organizations (MCOs) more than traditional health plans. This phenomenon has become known as "managed-care backlash." Using a model of the interaction between insurers, physicians, and patients, this paper identifies two possible motivations for MCO backlash. The first, which comes from traditional health plans' superior ability to credibly commit to providing better than least-cost care, is efficiency promoting. The second, which arises when patients are able to obtain higher-benefit treatments through reneging on their initial insurance contracts through "doctor shopping," may reduce efficiency. PMID- 16712988 TI - An ordered generalised extreme value model with application to alcohol consumption in Australia. AB - An Ordered Generalised Extreme Value (OGEV) model by Small (1987) is proposed for application to ordered discrete choice data. Relative to conventional Ordered Probit/Logit (OP/OL) and Multinomial Logit (MNL) models, the OGEV model is flexible, is defined by random utility maximization, and allows for correlation across choices via unobservable individual characteristics according to locations of the choices in the ordering. The OGEV model is applied to unit record data from Australia to study the impacts of prices, income and demographic characteristics on levels of alcohol consumption. Model selection analysis suggests that OGEV is preferred to both OP and MNL for the application. PMID- 16712989 TI - Development of a 9600-clone procedure for oligonucleotide fingerprinting of rRNA genes: utilization to identify soil bacterial rRNA genes that correlate in abundance with the development of avocado root rot. AB - Oligonucleotide fingerprinting of rRNA genes (OFRG) is an array-based method that generates microbial community profiles through analysis of rRNA gene clone libraries. The original OFRG method allowed 1536 clones to be analyzed per experiment. This report describes a procedure for analyzing 9600 clones per experiment, including a new probe set for bacterial analysis, and improved data processing and statistical analysis tools. The software tools are available at the OFRG website (). Use of the 9600-clone procedure was demonstrated by examining the bacterial rRNA gene compositions of soils subjected to various temperature treatments. These treatments produced a series of soils with a range of abilities to suppress avocado root rot, enabling the identification of bacterial rRNA genes that correlate in abundance with root rot suppressiveness. OFRG analysis of these soils produced 8876 bacterial rRNA gene fingerprints grouped into 5123 clusters, or operational taxonomic units (OTUs). Eleven OTUs exhibited a positive correlation between the number of clones and the percentage of healthy roots. An in silico analysis was performed to examine the relationship between the number of rRNA genes analyzed and the number of correlates (rRNA gene avocado root rot symptoms) identified. As the number of clones decreased, fewer correlates were identified. To further increase the throughput of the OFRG method, use of a glass slide-fluorescent probe microarray format was also explored. PMID- 16712990 TI - Improved expression and purification of the correctly folded response regulator PlnC from lactobacilli. AB - The response regulator PlnC is part of the signal transduction system that plays a key role in the regulation of bacteriocin production in Lactobacillus plantarum C11. In this study, we wanted to express high levels of the response regulator PlnC in a soluble and native form for purification and further studies. The protein was expressed as a fusion protein (fPlnC) containing an N-terminal Flag tag to facilitate detection and purification. When the fusion gene, fplnC, was expressed in Escherichia coli BL21, nearly all (99%) of the recombinant protein ended up inside inclusion bodies as an incorrectly folded protein. By utilizing two different Gram-positive expression systems (SIP and NICE) in L. plantarum NC8 and Lactobacillus sakei Lb790, the expression of the soluble fPlnC was significantly increased, being 20-40 times more than that in E. coli BL21. Using the N-terminal tag, the expressed protein was purified by immunoprecipitation. By DNA-binding study (EMSA), we demonstrated that the fusion protein purified from the soluble pool was correctly folded as judged by its ability to bind specifically on regulated promoters. Using our approach, we estimate that about 1 mg of fPlnC can be purified from 11 of the bacterial culture. PMID- 16712991 TI - Application of the quality index methodology for dosimetric verification of build up effect beyond air-tissue interface in treatment planning system algorithms. AB - We have designed a simple benchmark test for the user of a treatment planning system to check the calculation algorithm's ability to model the build up effect beyond an air/tissue interface. The expected result is expressed as an inhomogeneity correction factor CF derived from measurements and from Monte Carlo calculations for a full range of photon beam qualities. The linear regression lines obtained from plotting CF as a function of beam quality index form the basis for a quantitative check of the algorithm performance. PMID- 16712992 TI - Intensity-modulated radiotherapy of breast cancer using direct aperture optimization. AB - BACKGROUND AND PURPOSE: To design a clinically reliable and efficient step-and shoot IMRT delivery technique for the treatment of breast cancer using direct aperture optimization (DAO). Using DAO, segments are created and optimized within the same optimization process. PATIENTS AND METHODS: The DAO technique implemented in the Pinnacle treatment planning system, which is called direct machine parameter optimization (DMPO), was used to generate IMRT plans for twelve breast cancer patients. The prescribed dose was 50 Gy. Two DMPO plans were generated. The first approach uses DMPO only; the second technique combines DMPO with two predefined segments (DMPO(segm)), having shapes identical to the conventional tangential fields. The weight of these predefined segments is optimized simultaneously with DMPO. The DMPO plans were compared with normal two step (TS) IMRT, creating segments after optimizing the intensity. RESULTS: Dose homogeneity within the target volume was 4.8+/-0.6, 4.3+/-0.5 and 3.8+/-0.5 Gy for the TS, DMPO and DMPO(segm) plans, respectively. Comparing the IMRT plans with an idealized dose distribution obtained using only beamlet optimization, the degradation of the dose distribution was less for the DMPO plans compared with the two-step IMRT approach. Furthermore, this degradation was similar for all patients, while for the two-step IMRT approach it was patient specific. CONCLUSIONS: An efficient step-and-shoot IMRT solution was developed for the treatment of breast cancer using DMPO combined with two predefined segments. PMID- 16712993 TI - Finger pulse waveform length in the detection of concealed information. AB - An attempt was made to assess the efficiency of the finger pulse waveform length (FPWL), in detection of concealed information. For this purpose, two mock-theft experiments were designed. In the first, 40 guilty participants were examined while electrodermal, respiration and finger pulse volume were recorded. Results showed that detection accuracy with the FPWL was at least as good as the accuracy obtained with the other two measures (respiration changes and skin conductance responses). Detection efficiency was further improved when a combination of FPWL with the other two measures was used. In the second experiment, 39 guilty and 23 innocent participants were instructed to deny knowledge while the transducers were not attached to them. Then, the same questions were repeated while electrodermal, respiration and finger pulse volume were recorded. Results showed reduced rates of identification compared to the first experiment, which were explained by habituation. However, finger pulse was less affected by habituation than both respiration and skin conductance. Results suggested that the FPWL might be a useful addition to the existing measures in the detection of concealed information. PMID- 16712994 TI - Early gamma response of sleep is sensory/perceptual in origin. AB - The goal of the study was to investigate the gamma response of the brain and its functional correlates in rapid eye movements (REM) sleep and the three stages of non-REM sleep. Data on overnight sleep were acquired from 16 healthy, young adult, volunteer males. Neuroelectric activity was recorded from seven recording sites (Fz, Cz, Pz, F3, F4, P3, P4) in response to auditory stimuli (2000 Hz deviant and 1000 Hz standard stimuli: 65 dB, 10 ms r/f time, 50 ms duration) under passive oddball paradigm. Data were analyzed with the Fourier transform and digital filtering and also the recently developed technique of time-frequency component analysis (TFCA). TFCA displayed the gamma response under all stages of sleep. Statistical analysis did not reveal a significant effect of stimulus type, recording site or sleep stage on the three parameters of TFCA, which included maximum value of the time-frequency representation of the extracted gamma component, maximum magnitude of the time-domain representation of the component and the energy of this component. The gamma period included N1 and the early theta response, both of which are related to sensory-perceptual processing in the literature. According to these findings, the gamma response is possibly related, as in wakefulness, to early stimulus processing that also includes sensory/perceptual operations. PMID- 16712995 TI - Analysis of sensitivity to MK-801 treatment in a novel active allothetic place avoidance task and in the working memory version of the Morris water maze reveals differences between Long-Evans and Wistar rats. AB - The aims of the present study were to compare the effect of subchronic administration of MK-801 on performance in the active allothetic place avoidance (AAPA) task and in the working version of Morris water maze (MWM) in Long-Evans and Wistar rats. Animals were trained for four daily sessions either in the AAPA or in the working memory version of the MWM. Wistar rats treated by MK-801 (0.1 mg/kg) showed a cognitive deficit in the AAPA task without a significant hyperlocomotion, whereas they were not impaired in the working memory version of the MWM compared to controls. Long-Evans rats treated by MK-801 (0.1 mg/kg) were not impaired either in the AAPA task or in the MWM task. Higher doses of MK-801 (0.2 and 0.3 mg/kg) produced hyperlocomotion in both strains which corresponded to an inability to solve both spatial tasks. Long-Evans rats were superior in the MWM to the Wistar rats in the groups treated with the low dose of MK-801. In conclusion, intact Wistar rats can efficiently solve both spatial tasks; however, they are more sensitive to MK-801-induced behavioural deficit. This has relevance for modeling of the schizophrenia-related deficits and for screening substances for their therapeutic potential. PMID- 16712996 TI - Black aspergilli and ochratoxin A in grapes and wine. Introductory note. PMID- 16712997 TI - Genotyping of Campylobacter coli and C. jejuni from retail chicken meat and humans with campylobacteriosis in Slovenia and Bosnia and Herzegovina. AB - Thermotolerant Campylobacter jejuni and C. coli are one of the major causes of bacterial foodborne enteric infection. Consuming and/or handling poultry meat is the most consistent risk factor, linked to the high prevalence of campylobacters in retail poultry meat. The aim of the present study was to ascertain the genetic diversity and/or possible specificity of thermotolerant Campylobacter isolates according to species (C. coli, C. jejuni), isolation source (retail chicken meat and human clinical samples) and geographic origin (Goriska in Slovenia and Zenica Doboj Canton in Bosnia and Herzegovina (BIH)). With the pulsed-field gel electrophoresis (PFGE) after SmaI macrorestriction we distinguished 80 PFGE types among 118 strains and CfoI restriction fragment length polymorphism of the amplified flagellin gene (fla-RFLP) gave 12 fla-RFLP types. Beside the higher discriminatory power and strain typeability, PFGE discriminated the C. jejuni and C. coli groups of isolates. A high proportion of C. coli strains was isolated, especially from poultry samples. Identical or very similar PFGE types among the isolates from animal, food and human samples indicate the transmission of C. jejuni and C. coli from the chickens on the farm to the retail chicken meat, as well as possible cross-contamination of retail meat and transmission to humans. However, the identity of the isolates from non-related samples but with identical PFGE and fla-RFLP types should be confirmed with additional typing. Reliable tracing of the source of Campylobacter strains by molecular typing of the chicken meat isolates is therefore very difficult. The reasons include contamination of meat samples with multiple strains, possible cross-contamination and extreme heterogeneity of the isolates (mainly for C. jejuni) on one side and a limited power of the genotyping methods used to distinguish non-related strains on the other side (mainly for C. coli). PMID- 16712998 TI - European research on ochratoxin A in grapes and wine. AB - European wine production represents about 70% of world production and thus is an important export commodity. Ochratoxin A (OTA) was first detected as a wine contaminant in 1996 and the role of Aspergillus section Nigri and A. carbonarius in OTA production discovered in Europe in 1999. Subsequently Europe-wide surveys have shown that A. carbonarius is predominantly responsible for OTA contamination of grapes, wine and vine fruits. Analyses of wine samples throughout Europe have shown that there is a gradient in OTA concentration with a decrease from red, to rose and to white wines. The latitude of production is an important factor in determining risk from OTA contamination. Some geographic regions in Southern Europe are more prone to contamination with the toxigenic species and OTA. Ochratoxin A has also been found in much higher concentrations (max. 53 mug/kg) in dried vine fruit than in wine suggesting that A. carbonarius can dominate the drying vine fruit ecosystem. There is a significant lack of knowledge in Europe on conducive climatic conditions preharvest and their relationship with levels of risk from OTA contamination in grapes and their fate in wine production. This needs to be integrated with cultivation system to maximise the prevention of OTA entering this food chain. PMID- 16712999 TI - Flavoprotein monooxygenases, a diverse class of oxidative biocatalysts. AB - During the last decades a large number of flavin-dependent monooxygenases have been isolated and studied. This has revealed that flavoprotein monooxygenases are able to catalyze a remarkable wide variety of oxidative reactions such as regioselective hydroxylations and enantioselective sulfoxidations. These oxidation reactions are often difficult, if not impossible, to be achieved using chemical approaches. Analysis of the available genome sequences has indicated that many more flavoprotein monooxygenases exist and await biocatalytic exploration. Based on the known biochemical properties of a number of flavoprotein monooxygenases and sequence and structural analyses, flavoprotein monooxygenases can be classified into six distinct flavoprotein monooxygenase subclasses. This review provides an inventory of known flavoprotein monooxygenases belonging to these different enzyme subclasses. Furthermore, the biocatalytic potential of a selected number of flavoprotein monooxygenases is highlighted. PMID- 16713000 TI - Enhancement of pyruvate productivity in Torulopsis glabrata: Increase of NAD+ availability. AB - This study aimed at increasing the pyruvate productivity from a multi-vitamin auxotrophic yeast Torulopsis glabrata, by increasing the availability of NAD+. We examined two strategies for increasing availability of NAD+. To supplement nicotinic acid (NA), the precursor of NAD+; and to increase the activity of alcohol dehydrogenase integrating with addition acetaldehyde as exterior electron acceptor. The addition of 8 mg l(-1) NA to the fermentation medium resulted in a significant increase in the glucose consumption rate (48.4%) and the pyruvate concentration (29%). An ethanol-utilizing mutant WSH-13 was screened and selected after nitrosoguanidine mutagenesis of the parent strain T. glabrata CCTCC M202019. Compared with the parent strain, the alcohol dehydrogenase activity of the mutant WSH-13 increased about 110% and the mutant could utilize ethanol as the sole carbon source for growth (1.8 g l(-1) dry cell weight). When growing with glucose, the addition of 4 mg l(-1) acetaldehyde to the mutant WSH-13 culture broth led to a significant increase in the glucose consumption rate (26.3%) and pyruvate production (22.5%), but the ratio of NADH/NAD+ decreased to 0.22. Acetaldehyde did not affect the glucose and energy metabolism at high dissolved oxygen (DO) concentration. However, at lower DO concentration (20%), maintaining the acetaldehyde concentration in the mutant culture broth at 4 mg l( 1) caused an increased NAD+ concentration but a decreased NADH concentration. As a consequence, the pyruvate production rate, the pyruvate yield on glucose and the pyruvate concentration were 68, 44 and 45% higher, respectively, than the corresponding values of the control (without acetaldehyde). The strategy for increasing the glycolytic flux and the pyruvate productivity in T. glabrata by increasing the availability of NAD+ may provide an alternative approach to enhance the metabolites productivity in yeast. PMID- 16713001 TI - Oxygen defense in sulfate-reducing bacteria. AB - Sulfate-reducing bacteria (SRB) are strict anaerobes that are often found in biotopes where oxic conditions can temporarily exist. The bacteria have developed several defense strategies in order to survive exposure to oxygen. These strategies includes peculiar behaviors in the presence of oxygen, like aggregation or aerotaxis, and enzymatic systems dedicated to the reduction and the elimination of oxygen and its reactive species. Sulfate-reducing bacteria, and specially Desulfovibrio species, possess a variety of enzymes acting together to achieve an efficient defense against oxidative stress. The function and occurrence of these enzymatic systems are described. PMID- 16713002 TI - Cloning, purification and characterization of a functional anthracycline glycosyltransferase. AB - We have cloned the gene that encodes a novel glucosyl transferase (AraGT) involved in rhamnosylation of the polyketide antibiotic Aranciamycin in Streptomyces echinatus. AraGT comprises two domains characteristic of bacterial glycosyltranferases. AraGT was synthesized in E. coli as a decahistidinyl-tagged polypeptide. Purified AraGT is dimeric, displays a T(mapp) of 30 degrees C and can glycosylate the aglycone of an Aranciamycin derivative as shown by liquid chromatography and mass spectrometry. The availability of functional AraGT will allow the generation Aranciamycin-based combinatorial libraries. PMID- 16713003 TI - Functional expression of Candida antarctica lipase B in Eschericha coli. AB - Candida antarctica lipase B (CalB) is an important catalyst in bio-organic synthesis. To optimize its performance, either the reaction medium is changed or the lipase itself is modified. In the latter case, mutants are generated in Eschericha coli and subsequently expressed in fungal hosts for their characterization. Here we present the functional expression of CalB in the periplasm of E. coli. By step-wise deletion of the CalB signal and propeptide we were able to express and purify two different variants of CalB (mature CalB and CalB with its propeptide). A N-terminal FLAG and a C-terminal His tag were used for the purification. For the substrates para-nitrophenol butyrate (p-NPB), para nitrophenol laurate (p-NPL) and carboxyfluorescein diacetate (CFDA) the specific activity was shown to be similar to CalB expressed in Aspergillus oryzae. The kinetic constants k(M), v(max) and k(cat) were determined using the substrates p NPB and p-NPL. Almost identical k(cat)/k(M) values (0.423-0.466 min(-1) microM( 1) for p-NPB and 0.068-0.071 min(-1) microM(-1) for p-NPL) were obtained for the CalB variants from E. coli and A. oryzae. The results clearly show that CalB can be functionally expressed in E. coli and that the attachment of tags does not alter the properties of the lipase. PMID- 16713005 TI - Composite membrane of bacterially-derived cellulose and molecularly imprinted polymer for use as a transdermal enantioselective controlled-release system of racemic propranolol. AB - A composite membrane for transdermal delivery of S-propranolol enantiomer was developed based on the controlled pore functionalization of bacterial cellulose membranes using a molecularly imprinted polymer (MIP) layer synthesis. The reactive pore-filling of an asymmetric porous cellulose membrane with a MIP thin layer was effected using a silanized coupler as an additional anchor for the MIP. MIP thin-layers with specific binding sites for S-propranolol were synthesized by copolymerization of methacrylic acid with a cross-linker, ethylene glycol dimethacrylate in the presence of S-propranolol as the template molecule and the latter was subsequently extracted. Selective transport of S-propranolol through the MIP composite membrane was obtained, although this was determined mostly by the parent cellulose membrane with some ancillary contributory effect from the MIP layer. In addition, an enantioselectivity in the transport of propranolol prodrug enantiomers was found, suggesting that the shape and functional groups orientation, which are similar to that of the print molecule were essential for enantiomeric recognition of the MIP composite membrane. The enantioselectivity of S-MIP membranes was also shown when the release of propranolol enantiomers was studied in vitro using rat skin, with racemic propranolol contained in the donor compartment. The composite membrane of bacterially-derived cellulose and molecularly imprinted polymer may have great potential for use as a transdermal enantioselective controlled-release system for racemic propranolol. PMID- 16713004 TI - Molecular cloning, sequencing, expression of Chinese sturgeon cystatin in yeast Pichia pastoris and its proteinase inhibitory activity. AB - Cystatin, a superfamily of cysteine proteinase inhibitor of cathepsins and other cysteine proteinases, is widely distributed in animal tissues and body fluids. Although considerable attention has been given to mammalian and avian cystatins, little is known about cystatins from other vertebrates. In this study, a cDNA coding for Chinese sturgeon (Acipenser sinensis) cystatin was isolated and characterized. The corresponding mature cystatin peptide cDNA is 336 nucleotides long and encodes a protein of 112 amino acids. Sequence comparison showed that the cloned cystatin was a homolog of the mammalian Family II cystatin. The cystatin cDNA of Chinese sturgeon was subcloned into yeast expression vector pPICZalphaA and transformed into Pichia pastoris GS115 strain. After methanol induction, SDS-PAGE analysis of the culture supernatant indicated that the yield of recombinant cystatin was about 215 mg/l medium supernatant in shaking-flask fermentation medium, accounting for 73.6% of the total supernatant secreted proteins. Our data also showed that the recombinant cystatin is active in inhibiting the protease activity of papain and cathepsin B. Heat stability of the recombinant cystatin was also measured. PMID- 16713006 TI - [Relationship between walking capacity and maximal exercise capacity, strength and motor deficiency in adult hemiplegic stroke patients]. AB - OBJECTIVE: To evaluate the relation between walking capacity and maximal exercise capacity, strength and motor deficiency in hemiplegic stroke patients. DESIGN: Uncontrolled observational study. SETTING: Physical medicine and rehabilitation unit in a teaching hospital. PARTICIPANTS: Twenty hemiplegic stroke patients (17 men and 3 women) aged 18-70 years, whose stroke occurred more than 3 months before the study and who could walk independently with or without walking aids. MAIN OUTCOME MEASURES: Peak oxygen uptake (VO(2peak)), maximal power output walking capacity (6-min walk test) and motor deficiency (Fugl-Meyer scale). RESULTS: Walking capacity was correlated with both VO(2peak) and maximal power output (Pmax) [r=0.609, P<0.003 and r=0.868, P<0.0001, respectively] but also with strength (from r=0.640 to r=0.734; P=0.0018 to P=0.0001) and motor deficiency (r=0.6; P=0.004). CONCLUSIONS: Aerobic capacity and walking capacity are correlated and decreased in hemiplegic stroke patients. These results underscore the need for future studies to confirm the role of fitness in relation to walking capacity and to evaluate the benefit of integrating aerobic training into more traditional rehabilitation programs after stroke. PMID- 16713007 TI - Danggui-Buxue-Tang decoction has an anti-inflammatory effect in diabetic atherosclerosis rat model. AB - Danggui-Buxue-Tang (DBT) is a famous traditional Chinese formula. We examined the anti-inflammatory effect of it in diabetic atherosclerosis rats. DBT (3 or 6g/kg/day for 4 weeks) was orally administrated to the diabetic atherosclerosis rats, which were induced by nitric oxide inhibition (l-NAME in drinking water, 1mg/ml) plus high-fat diet. The concentrations of circulating inflammatory markers C-reactive protein (CRP) and tumour necrosis factor-alpha (TNF-alpha) and serum fructosamine were determined. The results showed that DBT had no direct effect in lowering serum fructosamine level, but can decrease the concentrations of CRP and TNF-alpha, produce a higher survival rate and less body weight loss, and decrease water intake in diabetic atherosclerosis in GK rats. PMID- 16713008 TI - 20-Hydroxyeicosatetraenoic acid is not associated with circulating insulin in lean to overweight humans. AB - Hypertension is often associated with insulin resistance or hyperinsulinemia. Metabolism of arachidonic acid by cytochrome P450 to the vasoconstrictor 20-HETE is thought to be involved in the pathogenesis of hypertension, but the relationship of 20-HETE with insulin resistance is not clearly understood. Obesity is a major determinant of insulin resistance, and we have previously demonstrated a positive relationship between BMI and 20-HETE in untreated hypertensive and normotensive individuals. The present analysis sought to determine if this relationship was related to insulin levels or insulin resistance (HOMA). We analysed 24h urinary excretion of 20-HETE, serum insulin levels and insulin resistance in 66 lean to overweight untreated hypertensive and normotensive individuals. There was a significant positive association between 20 HETE and BMI (p<0.001), and serum insulin and BMI (p=0.003). There were no associations between 20-HETE excretion and either serum insulin or insulin resistance, before or after adjustment for age, gender, BMI and BP, or when hypertensive and normotensive individuals were considered separately. The results of the present study suggest that the observed positive association between urinary 20-HETE excretion and BMI may not be related to circulating insulin levels or insulin resistance. PMID- 16713009 TI - The angiotensin II receptor antagonist valsartan enhances lipoprotein lipase mass in preheparin serum in type 2 diabetes with hypertension. AB - Recent studies suggest that blockade of angiotensin type 1 (AT1) receptor may have some effect on glucose and lipoprotein metabolism. Serum level of preheparin lipoprotein lipase (LPL) reflects LPL production mainly in adipocytes and is believed to be related to insulin sensitivity. We studied the effect of a selective AT1 antagonist, valsartan, on glucose, lipid metabolism and the preheparin LPL mass in 55 patients with type 2 diabetes and hypertension. Patients were randomized into a group administered valsartan 80 mg/day for 12 weeks or a group not administered valsartan (control). Blood pressure decreased significantly. HbA1c and TG levels decreased and HDL-C level increased, but these changes tended to be significantly different. TC and LDL-C levels were not significant changes. Preheparin LPL mass increased after valsartan administration compared with control (P = 0.0307), and migration ratio of LDL (LDL-Rm), which correlated negatively with LDL particle size, decreased compared with control (P < 0.0001). DeltaLDL-Rm correlated inversely with Delta preheparin LPL mass (r = 0.459). Among subjects treated with valsartan, greater improvement in preheparin LPL mass and blood pressure was observed in the subgroup with preheparin LPL mass <40 ng/ml. The results of this study suggest that valsartan may enhance LPL production in adipocytes, resulting in enlarged LDL particle size. PMID- 16713010 TI - Macro and microvascular complications are determinants of increased infection related mortality in Brazilian type 2 diabetes mellitus patients. AB - OBJECTIVE: To investigate infection-related mortality and its predictors in Brazilian type 2 diabetic patients. METHODS: It was carried out a long-term prospective study with 471 type 2 diabetic outpatients. Several clinical, laboratory and electrocardiographic variables were recorded at baseline. Predictive factors for infection-related mortality were evaluated by Kaplan-Meyer estimation of survival curves, univariate and multivariate Cox survival analysis. Excess infection-related mortality in this cohort was evaluated by comparing its rate with that of the Rio de Janeiro background population and calculating standardized mortality rates (SMR). RESULTS: During a median follow up of 57 months (range: 1-86 months), 40 (33.1%) patients died from infection-related causes. After adjusting for age and sex, the infection-related SMR was 6.6 (95% confidence interval [95% CI]: 4.8-9.0). In Cox multivariate analysis the predictors of infection-related mortality were older age (hazard ratio [HR]: 1.91; 95% CI: 1.35-2.70), pre-existing peripheral arterial disease (HR: 3.86; 95% CI: 1.80-8.28) and cerebrovascular disease (HR: 3.28; 95% CI: 1.24-8.70), lower HDL-cholesterol (HR: 2.50; 95% CI: 1.32-4.74) and increased 24h-proteinuria (HR: 1.22; 95% CI: 1.08-1.37). After excluding patients with peripheral and cerebrovascular disease at baseline, neuropathy and coronary heart disease were selected as predictors of mortality, besides older age and proteinuria. CONCLUSIONS: Brazilian type 2 diabetic patients have a six-fold excess infection related mortality than the general population. This increased mortality is mainly determined by the presence of micro and macrovascular complications. Multifactorial risk interventions are needed in order to decrease this burden of infection-related mortality. PMID- 16713011 TI - Clinical experience and applications of drug-eluting stents in the noncoronary vasculature, bile duct and esophagus. AB - OBJECTIVE: To review the use of drug-eluting stents outside the coronary artery. FINDINGS: The vast majority of research and clinical data on drug-eluting stents are from their use in coronary artery atherosclerosis; however, these devices can be used outside the coronary circulation in both vascular and nonvascular structures. In noncoronary arteries the principle indication for drug-eluting vascular stents is the same as in the coronary circulation, prevention of restenosis. Human experience has been essentially limited to trials or compassionate use; two small controlled studies and a number of small observational single center reports have been published, and there are trials in progress. To date the data have not been as compelling as in the coronary circulation. The physical characteristics of each vascular bed such as external compressive forces, blood flow rates, wall thickness relative to lumen size, and vessel wall composition differ significantly from the coronary circulation and each presents unique challenges to local drug delivery. Outside the vascular bed, the principle expected use is the prevention of tissue ingrowth after stent insertion in tubular structures such as the trachea, esophagus or bile ducts. CONCLUSIONS: Considerable further study of drug-eluting stents will be required in each anatomic region to determine the ideal stent/drug combination and clinical appropriateness. PMID- 16713012 TI - Effects of different combinations of gefitinib and irinotecan in lung cancer cell lines expressing wild or deletional EGFR. AB - EGFR mutations are a major determinant of lung tumor response to gefitinib, an EGFR-specific tyrosine kinase inhibitor. Obtaining a response from lung tumors expressing wild-type EGFR is a major obstacle. The combination of gefitinib and cytotoxic drugs is one strategy against lung cancers expressing wild-type EGFR. The DNA topoisomerase inhibitor irinotecan sulfate (CPT-11) is active against lung cancer. We examined the sensitivity of lung cancers expressing wild- or mutant-type EGFR to the combination of gefitinib and CPT-11. The in vitro effect of gefitinib and SN-38 (the active metabolite of CPT-11) was examined in seven lung cancer cell lines using the dye formation assay with a combination index. When administered concurrently, gefitinib and SN-38 had a synergistic effect in five of the seven cell lines expressing wild-type EGFR, whereas the combination was antagonistic in PC-9 cells and a PC-9 subline resistant to gefitinib and expressing deletional mutant EGFR (PC-9/ZD). When administered sequentially, treatment with SN-38 followed by gefitinib had remarkable synergistic effects in the PC-9 and PC-9/ZD cells. In an in vivo tumor-bearing model, this combination had a schedule-dependent synergistic effect in the PC-9 and PC-9/ZD cells. An immunohistochemical analysis of the tumors in mice treated with CPT-11 and gefitinib demonstrated that the number of Ki-67 positive tumor cells induced by CPT-11 treatment was decreased when CPT-11 was administered in combination with gefitinib. In conclusion, the sequential combination of CPT-11 and gefitinib is considered to be active against lung cancer. PMID- 16713014 TI - Is a healthy ecosystem one that is rich in parasites? AB - Historically, the role of parasites in ecosystem functioning has been considered trivial because a cursory examination reveals that their relative biomass is low compared with that of other trophic groups. However there is increasing evidence that parasite-mediated effects could be significant: they shape host population dynamics, alter interspecific competition, influence energy flow and appear to be important drivers of biodiversity. Indeed they influence a range of ecosystem functions and have a major effect on the structure of some food webs. Here, we consider the bottom-up and top-down processes of how parasitism influences ecosystem functioning and show that there is evidence that parasites are important for biodiversity and production; thus, we consider a healthy system to be one that is rich in parasite species. PMID- 16713015 TI - MRSA behind bars? PMID- 16713013 TI - Efficient palliation in patients with small-cell lung cancer by a combination of paclitaxel, etoposide and carboplatin: quality of life and 6-years'-follow-up results from a randomised phase III trial. AB - PURPOSE: Based on the promising activity of paclitaxel in small-cell lung cancer (SCLC) we conducted a randomized phase III trial to evaluate whether a combination of paclitaxel, carboplatin and etoposide phosphate (TEC) improves survival and time to progression as well as tolerability and quality of life (QoL) compared to a regimen of carboplatin, etoposide phosphate and vincristine (CEV) in SCLC patients. PATIENTS AND METHODS: Six hundred and fourteen patients with stages I-IV SCLC were randomly assigned between January 1998 and December 1999 to both treatment arms. All patients were evaluated for response rate, survival, side effects and quality of life with overall survival (OS) serving as primary endpoint. A final analysis was done after a six-year follow-up. Survival curves were estimated using Kaplan-Meier curves and tested with the log-rank test. Quality of life data were assessed in using the EORTC QLQ-C30 questionnaire and evaluated by calculating and comparing the mean scores as well as applying longitudinal techniques. RESULTS: Six hundred and eight patients were evaluable for efficacy and toxicity. The long-term follow-up confirms the significant survival benefit for the paclitaxel, etoposide, carboplatin (TEC) regimen with a median OS of 12.5 months compared to 11.7 months for the CEV arm (HR, 1.21; 95% CI, 1.02-1.43; P=.030). The 5-year survival rates were 14% for the experimental versus 6 % for the CEV arm. Significant survival prolongation was also observed in the subgroup of patients with stage IV disease (HR, 1.27; 95% CI, 1.00-1.60; P=.047). The previously reported clinical benefit in form of an overall reduction of grade 3/4 toxicity was backed by the results of the comprehensive QoL analysis we report hereby. TEC significantly improves the relevant QoL parameters like global overall QoL or physical functioning. CONCLUSION: When administered in combination with etoposide and carboplatin, paclitaxel is able to offer in SCLC patients with extensive disease a survival benefit without additional toxicities, but with gains in patient-reported quality of life. In terms of efficient palliative care, TEC might be seen as an alternative to standard cisplatin plus etoposide in patients requesting a powerful palliative regimen not compromising any survival benefit. PMID- 16713016 TI - Emerging plasmid-mediated quinolone resistance associated with the qnrA gene in Enterobacter cloacae clinical isolates in China. PMID- 16713017 TI - Hand hygiene practices among ophthalmologists. PMID- 16713018 TI - Staphylococcus aureus bacteraemia: incidence, risk factors and predictors for death in a Brazilian teaching hospital. AB - The aim of this study was to evaluate the incidence of bloodstream infection due to Staphylococcus aureus and the risk factors for mortality. The design was a two year retrospective cohort of patients more than one year of age with clinically significant and microbiologically documented bloodstream infection due to S. aureus between January 2000 and December 2001 in a tertiary teaching hospital in midwest Brazil. One hundred and eleven patients were identified with clinically significant and microbiologically confirmed bacteraemia due to S. aureus, accounting for an infection rate of five per 1000 admissions. Nosocomial infections represented 83.8% of cases and meticillin-resistant Staphylococcus aureus (MRSA) accounted for 60.2% of cases. Overall mortality due to S. aureus bacteraemia was 35.1%. Infection due to MRSA, severity of clinical status (severe sepsis or septic shock) and inadequate initial antimicrobial therapy were identified by univariate analysis as predictors of mortality. After Cox regression analysis, severity of clinical manifestations [hazard ratio (HR) 6.86, 95% confidence interval (CI) 3.05-15.43] and inadequacy of antimicrobial therapy (HR 2.27, 95%CI 1.02-5.09) remained as risk factors for mortality. Early diagnosis of bacteraemia should be sought in order to implement adequate treatment before the onset of severe sepsis and septic shock, thus reducing the mortality rate. PMID- 16713019 TI - Nosocomial aspergillosis in outbreak settings. AB - Nosocomial aspergillosis represents a serious threat for severely immunocompromised patients and numerous outbreaks of invasive aspergillosis have been described. This systematic review summarizes characteristics and mortality rates of infected patients, distribution of Aspergillus spp. in clinical specimens, concentrations of aspergillus spores in volumetric air samples, and outbreak sources. A web-based register of nosocomial epidemics (outbreak database), PubMed and reference lists of relevant articles were searched systematically for descriptions of aspergillus outbreaks in hospital settings. Fifty-three studies with a total of 458 patients were included. In 356 patients, the lower respiratory tract was the primary site of aspergillus infection. Species identified most often were Aspergillus fumigatus (154 patients) and Aspergillus flavus (101 patients). Haematological malignancies were the predominant underlying diseases (299 individuals). The overall fatality rate in these 299 patients (57.6%) was significantly greater than that in patients without severe immunodeficiency (39.4% of 38 individuals). Construction or demolition work was often (49.1%) considered to be the probable or possible source of the outbreak. Even concentrations of Aspergillus spp. below 1 colony forming unit/m(3) were sufficient to cause infection in high-risk patients. Virtually all outbreaks of nosocomial aspergillosis are attributed to airborne sources, usually construction. Even small concentrations of spores have been associated with outbreaks, mainly due to A. fumigatus or A. flavus. Patients at risk should not be exposed to aspergilli. PMID- 16713020 TI - Persistent colonization and genetic diversity of Helicobacter pylori among patients with severe neurological impairment. PMID- 16713021 TI - The risk of disease transmission associated with inadequate disinfection of gastrointestinal endoscopes. PMID- 16713023 TI - Hemopressin, a hemoglobin fragment, dilates the rat systemic vascular bed through release of nitric oxide. AB - The present study was undertaken to investigate the effects of intravenous (i.v.) administration of rat hemopressin (rHP), 30-1000 microg/kg, on systemic arterial pressure (SAP), cardiac output (CO) and systemic vascular resistance (SVR) in the anesthetized rat. Bolus i.v. injections of rHP produced mild decreases in SAP that were dose-dependent. Since CO was not altered, the decreases in SAP reflect reductions in SVR. The systemic vasodilator response to rHP was not subject to tachyphylaxis. The systemic vasodilator response to rHP was abolished by L-nitro arginine methylester (L-NAME) but was not altered by meclofenamate. In addition, rHP lacked direct contractile and relaxant activity on isolated rat aortic rings (AA) and pulmonary arterial rings (PA). The present data suggest rHP dilates the rat systemic vascular bed through the endogenous release of nitric oxide (NO) independent of the formation of cyclooxygenase products including prostacyclin. It is possible rHP acts as an endogenous vasodilator substance to regulate local blood flow during clinical states of altered red cell turnover, microvascular disease and hemolysis. PMID- 16713025 TI - Plasmodium falciparum merozoite surface protein 6 (MSP-6) derived peptides bind erythrocytes and partially inhibit parasite invasion. AB - This work shows that Plasmodium falciparum merozoite surface protein-6 (MSP-6) peptides specifically bind to membrane surface receptor on human erythrocytes. Three high activity binding peptides (HABPs) were found: peptides 31175 (41MYNNDKILSKNEVDTNIESN60) and 31178 (101YDIQATYQFPSTSGGNNVIP120) in the amino terminal region and 31191 (361EIDSTINNLVQEMIHLFSNNY380) at the carboxy terminal. Their binding to erythrocytes was saturable. HABPs 31191 and 31178 recognized 56 and 26 kDa receptors on erythrocyte membrane and inhibited in vitro Plasmodium falciparum merozoite invasion of erythrocytes by between 27% and 46% at 200 microg ml(-1) concentration, suggesting that these MSP-6 protein peptides play a possible role in the invasion process. PMID- 16713026 TI - Synthesis and biological evaluation of new vinyl ester pseudotripeptide proteasome inhibitors. AB - Here we report the synthesis and biological activities of new tripeptidic-based vinyl ester derivative proteasome inhibitors. Starting from Hmb-Val-Ser-Leu-VE prototype, we investigated P2 position and N-terminal substitution. The more effective inhibitors of the series showed remarkable inhibition and selectivity for the trypsin-like (beta2) subunit and were revealed to be specific for the proteasome. In vitro metabolic stability studies of the new vinyl ester analogues are also reported here. PMID- 16713027 TI - [Recent advances in the treatment of myelodysplastic syndromes]. AB - OBJECTIVES: Myelodysplastic syndromes (MDS) are a group of clonal hematologic disorder, which combine ineffective hematopoiesis and evolution to acute myeloid leukemia. Significant progress has been made in the understanding of the disease pathogenesis, diagnostics and classification. Promising new agents and innovative therapeutic strategies are currently used. In this article we will review these achievements and their impact on the treatment of MDS. CURRENT KNOWLEDGE AND KEY POINTS: The pathogenesis of MDS involves abnormalities of the MDS clone itself such as abnormal apoptosis, signalling or epigenetic regulation and abnormalities of the microenvironment such as immune deregulation or increased angiogenesis, which represent potential therapeutic targets. There is currently no standard treatment for MDS and allogeneic stem cell transplantation remains the only curative strategy. However, besides conventional chemotherapy and growth factors, new agents including hypomethylating agents, antiangiogenic drugs, immune modulatory agents have proved effective. FUTURE PROSPECTS AND PROJECTS: The interesting results achieved with these new agents show that it is necessary to continue investigation in order to improve therapeutic strategies in MDS. PMID- 16713028 TI - [New definition of optimal vitamin D status and redefening serum parathyroid hormone reference range]. AB - SCOPE: Knowledge concerning vitamin D has greatly improved during the past few years. Vitamin D can no longer be considered only as a preventive therapy for rickets-osteomalacia. Indeed, beside its role in the prevention of osteoporotic fractures in the elderly, many data suggest that it may be involved in the prevention of various diseases including cancers and auto-immune diseases. CURRENT SITUATION AND SALIENT POINTS: Vitamin D status may be easily assessed by the measurement of 25OHD serum concentration. However, many specialists in the field regard most 25OHD reference values as being too low, and believe that the 25OHD threshold below which vitamin D status can be considered as insufficient is somewhere between 50 and 100 nmol/L (20 to 40 ng/mL). It then appears that usually recommended amounts of supplemental vitamin D may be too low to reach these 25OHD concentrations, and thus need to be revised. We have proposed that PTH reference values should be established in healthy subjects with a normal vitamin D status. This supposes that 25OHD is measured in the reference population beforehand, and that the subjects with vitamin D insufficiency are eliminated from the reference group. PERSPECTIVES: Although more complicated than the usual way to establish normative data, we have shown that it decreases the upper limit of normal by 25-35%, enhancing thus the diagnostic sensitivity for hyperparathyroidism without a decrease in specificity. PMID- 16713029 TI - [Aortic abdominal aneurysm rupture leading to a massive gastrointestinal bleeding in a patient with Behcet's disease]. AB - INTRODUCTION: Arterial involvement in Behcet's disease is rare, present in 2-8% of cases. Aortic aneurysms represent the most frequent arterial lesions encountered in this disease and are associated with high mortality. CASE REPORT: We report the exceptional observation of an abdominal aortic aneurysm ruptured to the duodenum, responsible of a massive gastro intestinal bleeding in a patient with Behcet's disease requiring an emergent surgical intervention. The outcome was favorable. DISCUSSION: The clinical presentation of abdominal aortic aneurysms in Behcet's disease is often atypical leading to an important diagnosis delay and favours the rupture of the aneurysm. PMID- 16713030 TI - Establishing a culture for patient safety - the role of education. AB - This paper argues that the process of making significant moves towards a patient safety culture requires changes in healthcare education. Improvements in patient safety are a shared international priority as too many errors and other forms of unnecessary harm are currently occurring in the process of caring for and treating patients. A description of the patient safety agenda is given followed by a brief analysis of human factors theory and its use in other safety critical industries, most notably aviation. The all too common problem of drug administration errors is used to illustrate the relevance of human factors theory to healthcare education with specific mention made of the Human Factors Analysis and Classification System (HFACS). PMID- 16713031 TI - Intestinal permeability and oxidative stress in patients with alcoholic pellagra. AB - BACKGROUND & AIMS: Increased intestinal permeability is one of the grastointestinal changes observed in alcoholic patients. However, there are no objective definitions as yet of how alcohol induces pathological changes in the various organs. The action of oxygen-free radicals during ethanol metabolism has been considered a determinant factor of these alterations. The present study was undertaken to determine the effect of niacin supplementation on intestinal permeability and oxidative stress in patients with alcoholic pellagra. METHODS: The study was divided into two phases: in Phase 1 we studied ten patients with pellagra before treatment with niacin, and in Phase 2 we studied the same patients after 27 days of treatment with niacin. Intestinal permeability was assessed by the (51)CrEDTA test and the antioxidant action of niacin supplementation was assessed by the determination of lipid peroxidation (plasma malondialdehyde, MDA), protein oxidation (plasma carbonyl group) and of the antioxidants plasma vitamin E and erythrocyte glutathione peroxidase. RESULTS: Comparison of intestinal permeability by the (51)CrEDTA test before and after niacin treatment showed a significant decrease in permeability from 4.29+/-1.92% to 1.90+/-1.19% (P<0.05). Assessment of oxidative stress showed a significant decrease (P<0.05) in lipid and protein peroxidation (MDA: 1.19+/-0.40-0.89+/-0.27 micromol/l; carbonyl groups: 2.22+/-0.36-1.84+/-0.40 nmol/mg protein). CONCLUSIONS: The results suggest that niacin and vitamin E deficiency in patients with pellagra could be important factors in increased intestinal permeability and decreased antioxidant conditions, recovering to normal values after treatment with niacin, associated to alcohol abstinence and a balanced diet. PMID- 16713033 TI - Multigenotype HCV-NS3 recombinant vaccinia viruses as a model for evaluation of cross-genotype immunity induced by HCV vaccines in the mouse. AB - Surrogate infections with HCV-recombinant vaccinia viruses (HCV-rVV) are a standard method to test the efficacy of hepatitis C virus (HCV) vaccine candidates in the mouse model. We established a panel of 16 HCV-rVV expressing the nonstructural protein 3 (NS3) of HCV genotypes 1a, 1b, 2, 3 and 4. Mice immunized with recombinant NS3 protein derived from HCV genotype 1b were challenged with the rVV. rVV-titers decreased up to 54-fold after subtype 1b challenge and up to 8.5-fold after subtype 1a challenge. No change was detected for genotype 2, 3, or 4. Our model is a convenient and reliable tool to analyze the induction of cross-genotype immunity by experimental vaccination of mice. PMID- 16713032 TI - The HIV-1 matrix protein p17 can be efficiently delivered by intranasal route in mice using the TLR 2/6 agonist MALP-2 as mucosal adjuvant. AB - The HIV-1 matrix protein p17 is a structural protein essential in the life cycle of HIV, by acting as a virokine/immunomodulator that supports viral replication and spreading. The presence of p17-specific antibodies and CTL responses correlates with slower progression to AIDS. Intranasal vaccination with p17 and the TLR2/6 agonist MALP-2 stimulates strong humoral and cellular immune responses at systemic and mucosal levels. The antibodies blocked p17 binding to its receptor, which is a critical step for the exertion of its virokine activity. Our results suggest that p17 and MALP-2 are attractive candidates for incorporation in mucosal vaccines against HIV/AIDS. PMID- 16713034 TI - Efficacy of strain RB51 vaccine in heifers against experimental brucellosis. AB - With the goal of providing an additional tool for controlling bovine brucellosis in Brazil and evaluating the full calf dose in adult cattle, the efficacy of the rough Brucella abortus strain RB51 vaccine was tested in heifers. Thirty-three females of approximately 24 months of age were divided in two groups: one group (n=20) received the RB51 vaccine and the other group (n=13) were used as non vaccinated control. Animals in the vaccinated group were split in two sub-groups. One sub-group (n=12) was vaccinated subcutaneously with 1.5x10(10) colony forming units (CFU) of RB51 at Day 0 of the experiment and the other sub-group (n=8) was vaccinated subcutaneously with 1.6x10(10) CFU of RB51 at 60 days of gestation (Day 260 of the experiment). All cattle were challenged between 6 and 7 months of pregnancy with 3x10(8) CFU of the virulent strain 2308 of B. abortus by the conjunctival route. Vaccination with RB51 vaccine did not result in the production of any antibodies against the O-side chain of lipopolysaccharide (LPS), as measured by conventional serological tests (rose bengal plate agglutination test (RBPAT), standard tube agglutination test (STAT), and 2 mercaptoethanol test (2ME)). A total of 25% cumulative incidence of abortions was found in the vaccinated group, whereas in the control group the cumulative incidence was 62%. B. abortus RB51 was not isolated from any sample, and no abortions were produced by RB51 vaccination of females at 60 days of pregnancy. The results indicate that vaccination with RB51 prevented 59.4% of abortions, 58.6% of cow infections, and 61.0% of fetal infections. The relative risk (RR) revealed that non-vaccinated animals have 2.462 (95% CI 1.029-5.889) times higher risk of aborting than RB51-vaccinated animals. PMID- 16713035 TI - Development of hepatitis B oral vaccine using B-cell epitope loaded PLG microparticles. AB - Oral hepatitis B vaccine formulation was prepared by successful encapsulation of immunogenic peptide representing residues 127-145 of the immunodominant B-cell epitope of hepatitis B surface antigen (HBsAg) in poly(D,L-lactide co-glycolide) (PLG) microparticles. The smooth, spherical PLG microparticles with a diameter of around 10 microm was prepared by using W/O/W double emulsion solvent evaporation method. The entrapment efficiency of B-cell epitope peptide (BCEP) into PLG microparticles was 64%. In vitro studies showed B-cell epitope loaded PLG microparticles (BCEM) released the peptide in sustained profile and reached 64.9% efficiency by Day 25. Single oral immunization of mice with BCEM led to the significant induction of specific serum IgG and IgM anti-HB antibodies. After the termination of antibody induction, the orally immunized mice were infected with HBsAg, which resulted in the rapid production of antibodies against HBsAg as a result of secondary immune response. PLG microparticles formulation approach may have potential in increasing the efficacy of microparticulate systems for the oral administration of hepatitis B vaccine. PMID- 16713036 TI - Re: Natural infection with Japanese encephalitis virus among inhabitants of Japan: a nationwide survey of antibodies against nonstructural 1 protein. AB - Investigations to assess episodes of any natural Japanese encephalitis (JE) infection following prior immunizations with JE inactivated vaccine in Japan are commendable. Employing differential production of nonstructural 1 protein NS1, during a natural infection and not by inactivated JE vaccine [Konishi E, Shoda M, Yamamoto S, Arai S, Tanaka-Taya K, Okabe N. Natural infection with Japanese encephalitis virus among inhabitants of Japan: a nationwide survey of antibodies against nonstructural 1 protein. Vaccine 2006;24:3054-956], natural infection was manifest in eight selected prefectures in Japan. Moreover, it would be possible to detect any foreign pathogenic JE virus (JEV) or any native strains associated with any major shift in JE clinical presentations. On the opposite, any NS1 antibody screen might be invalid among recipients of live JE vaccines. PMID- 16713037 TI - Protection against H1, H5, H6 and H9 influenza A infection with liposomal matrix 2 epitope vaccines. AB - The recent emergence of multiple avian influenza A subtypes that cause human disease (i.e., H5N1, H9N2 and H7N7), coupled with the fear that one of these strains might precipitate a new pandemic, underscores the need to develop new technological approaches to immunization which elicit protective immune responses against multiple subtypes of influenza A. In response to this demand, several matrix 2 protein ectodomain segments (M2eA) corresponding to the H1N1, H5N1 and H9N2 influenza strains were formulated using a novel liposome-based vaccine technology and evaluated as potential immunogens for developing a "universal" influenza vaccine. Mice immunized with liposomal M2eA survived homologous challenges with H1N1 (100% survival) or H9N2 (80% survival) influenza strains. There were significant reductions in their lung viral load as well as in immunized mice challenged with the H5N1 subtype. The mice vaccinated with an M2eA segment corresponding to the H1N1 and H6N2 (a reassortant influenza A virus carrying the M2eA from PR8/34) strains elicited elevated IgG ELISA antibody titers to this M2eA epitope segment and antiserum from these immunized mice provided passive protection (100% survival) to naive mice receiving a lethal dose of H6N2 influenza virus. These results provide the first evidence that recombinant M2eA epitopes to multiple subtypes elicited immune protection against a homologous challenge and provides further evidence in favor of the development of a "universal" influenza vaccine based on M2eA. PMID- 16713038 TI - Use of National Health Insurance database to evaluate the impact of public varicella vaccination program on burden of varicella in Taiwan. AB - The aims of this study were to investigate the epidemiology of varicella and evaluate the effect of varicella vaccination on the burden of varicella in Taiwan. The outpatient and hospitalization claims made to the National Health Insurance (NHI) in public and private vaccination areas were compared during 2000 2002. In 2002, the outpatient visit rates for 1- to 5-year-old children in public vaccination areas were 66-78% lower than their counterparts in private vaccination areas. The reduction in outpatient visit rate was the greatest among preschool children. The reductions also occurred in every age group including infants and adults. This study showed significant evidence of vaccine impact with a marked decline in clinical cases in the short term. PMID- 16713039 TI - Contamination of terrestrial gastropods, Helix aspersa maxima, with 137Cs, 85Sr, 133Ba and 123mTe by direct, trophic and combined pathways. AB - (137)Cs, (85)Sr, (133)Ba and (123m)Te contaminations of terrestrial gastropods, Helix aspersa maxima, by direct deposition, labelled food ingestion or combined (trophic and direct pathways) exposure were carried out under laboratory conditions. The aim of this study was to compare the three contamination pathways: direct, trophic and combined, in terms of individual mortality, radionuclide uptake, depuration and distribution in the tissues. An initial group of 30 snails (2 years old) was exposed to radioactive aerosols during a 20-h period. These aerosols were assumed to be representative of those that would be released during a nuclear accident occurring in a PWR. A second group of 50 snails (same age) was submitted to an ingestion of commercial food contaminated by the same aerosols, twice a week for 21 days (flour at a feeding rate of about 0.2g). A third group of 40 snails was submitted to a combined exposure: exposure to radioactive aerosols (20h), followed by ingestion of flour contaminated by the same aerosols, twice a week for 21 days. No significant difference between the three groups and a reference group of 10 snails was observed, neither in growth nor in mortality. Concerning the direct pathway, at the end of direct deposition (about 1 day after the beginning), cesium was the most bioavailable element, distributed rather homogeneously throughout the whole body (13% of the total Cs in all organs excepting the digestive system and 28% in the muscle). Strontium was measured in the shell (about 70%). Barium was found in the muscle (20%) and in the shell (65%). Tellurium was mainly present in the shell (70%) and in the digestive system (20%). After 21 days of depuration, the faeces eliminated 42% of the Te. As for contamination by ingestion, Te mainly accumulated in the digestive system (72% of Te present in the total body), Ba accumulated in the muscle (75%) and Sr in the shell (70%). Concerning contamination by combined pathways, at the end of the 21-day exposure, the 4 radionuclides had the same tendency as direct deposition. However, the effect of the trophic pathway was significant: it causes an 18% increase of Sr in the shell and an 7% increase of Cs in the digestive system in comparison to direct deposition, resulting in a final 86% in the shell and 27% in the digestive system. PMID- 16713040 TI - Evaluation of substitutes for bone: comparison of microradiographic and histological assessments. AB - We created defects of standard size in the frontal bones of adult pigs and filled them with four different materials. On six occasions (at 1, 2, 4, 8, 12, and 26 weeks), samples were harvested, and evaluated by computing microradiographic images. We examined the specimens histologically as controls. After insertion of anorganic materials, microradiographic evaluation was easy and precise, and there were no significant differences between them and the histological controls (p=0.2). A quantitative evaluation of chemically sterilised bone by computer was not possible for more than 4 weeks. PMID- 16713041 TI - Injuries to the head and neck in Homer's Iliad. AB - We have found and reported on all injuries to the head and neck described in Homer's Iliad, and give several particularly graphic examples. PMID- 16713042 TI - DNA analysis of the SH3BP2 gene in patients with aggressive central giant cell granuloma. AB - A mutation of the SH3BP2 gene is known to cause cherubism. As there are clinical and histopathological similarities between central giant cell granuloma and cherubism, we made a constitutional DNA analysis of the SH3BP2 gene in four patients with aggressive giant cell granuloma (having one or more of the following features pain, paraesthesia, rapid growth, or root resorption). We found no mutations in the SH3BP2 gene, which indicates that cherubism is a separate entity. However, a somatic mutation in a specific group of cells could cause the focal lesions in giant cell granuloma. Further DNA analysis of the tissue of giant cell granulomas therefore seems indicated. PMID- 16713043 TI - Interesting case: Painless massive swelling in the oral cavity. PMID- 16713044 TI - Post-traumatic eye observations M.C. Bater, P.L. Ramchandani, P.A. Brennan, Br. J. Oral Maxillofac. Surg. 43 (2005) 410-416. PMID- 16713045 TI - 'Nesting' and 'Matrescence' as distinctive features of a free-standing birth centre in the UK. AB - OBJECTIVE: To explore the culture, beliefs, values, customs and practices around the birth process within a free-standing birth centre (FSBC). DESIGN: Ethnography. SETTING: A birth centre situated in the midlands of England. PARTICIPANTS: Women attending the centre, midwives and maternity-care assistants (MCAs) working at the centre. FINDINGS: Women in the study seemed to invoke intuitive nesting-related behaviours in their assessment of the suitability of the birth centre. In addition, the birth centre staff's focus on creating the right ambience for birth may also emanate from nesting concerns. Birth-centre staff assisted women through the 'becoming mother' transition, which is conceptualised as 'matrescent' care. KEY CONCLUSIONS: The birth-centre environment elicited nesting-like behaviours from both women and staff. This formed part of a nurturing orientation that was conceptualised as 'matrescent' (becoming mother) care. 'Matrescence' does not seem to be grounded in clinical skills but is relationally mediated. IMPLICATIONS FOR PRACTICE: Nesting-like behaviours and 'matrescent' care in this context challenge maternity services to review traditional conceptualisations of safety and traditional expressions of clinical intrapartum care. PMID- 16713046 TI - A comparison of the viscoelastic properties of bone grafts. AB - BACKGROUND: The expansion in joint arthroplasty surgery in the 1970s has resulted in a large group of patients who require revision arthroplasty for aseptic loosening. Impaction bone grafting to deal with bone stock loss has become an increasingly popular procedure in revision hip surgery. The results of this revision surgery are variable and very much dependent on the grafting techniques adopted at the time of surgery. In vitro testing of impaction bone grafting methods can constitute an important tool to improve long-term clinical results. The increasing clinical demand for human allograft limits its availability for use in in vitro laboratory studies therefore suitable experimental alternatives are required. METHODS: Human, porcine and ovine cancellous bone grafts were morsellised and prepared for in vitro laboratory testing following a standard operative technique. Each graft type was compressed using a die plunger test. After compression to a predetermined level the graft was left to relax for 120s. A comparison of the compression moduli and of the relaxation characteristics for each graft preparation was performed. In addition, the effects of washing the graft and of cartilage removal from the graft mixture were investigated. RESULTS: This study has demonstrated that there is no statistical difference in the compression modulus or relaxation percentage between human and ovine graft preparations. The effect of removing cartilage and washing the graft mixtures were inconsistent with regard to alterations in the viscoelastic properties of the grafts. INTERPRETATION: On the basis of the experiments performed we recommend the use of ovine bone graft as a suitable substitute for human allograft for in vitro testing of impaction bone grafting methods. The properties of ovine graft were similar for both compression moduli and relaxation properties to human allograft. PMID- 16713047 TI - Large variation in lipid content, SigmaPCB and delta13C within individual Atlantic salmon (Salmo salar). AB - Many studies that investigate pollutant levels, or use stable isotope ratios to define trophic level or animal origin, use different standard ways of sampling (dorsal, whole filet or whole body samples). This study shows that lipid content, SigmaPCB and delta(13)C display large differences within muscle samples taken from a single Atlantic salmon. Lipid- and PCB-content was lowest in tail muscles, intermediate in anterior-dorsal muscles and highest in the stomach (abdominal) muscle area. Stable isotopes of carbon (delta(13)C) showed a lipid accumulation in the stomach muscle area and a depletion in tail muscles. We conclude that it is important to choose an appropriate sample location within an animal based on what processes are to be studied. Care should be taken when attributing persistent pollutant levels or stable isotope data to specific environmental processes before controlling for within-animal variation in these variables. PMID- 16713048 TI - Ozone induced leaf loss and decreased leaf production of European Holly (Ilex aquifolium L.) over multiple seasons. AB - European Holly (Ilex aquifolium L.) was used to study the impact of one short (28 day) ozone fumigation episode on leaf production, leaf loss and stomatal conductance (g(s)), in order to explore potential longer term effects over 3 growing seasons. Young I. aquifolium plants received an episode of either charcoal-filtered air or charcoal-filtered air with 70 nl l(-1) O(3) added for 7 h d(-1) over a 28 day period from June 15th 1996, then placed into ambient environment, Stoke-on-Trent, U.K. Data were collected per leaf cohort over the next three growing seasons. Ozone exposure significantly increased leaf loss and stomatal conductance and reduced leaf production over all subsequent seasons. Impact of the initial ozone stress was still detected in leaves that had no direct experimental ozone exposure. This study has shown the potential of ozone to introduce long-term phenological perturbations into ecosystems by influencing productivity over a number of seasons. PMID- 16713049 TI - Comparison of techniques for estimating PAH bioavailability: uptake in Eisenia fetida, passive samplers and leaching using various solvents and additives. AB - The aim of this study was to evaluate different techniques for assessing the availability of polycyclic aromatic hydrocarbons (PAHs) in soil. This was done by comparing the amounts (total and relative) taken up by the earthworm Eisenia fetida with the amounts extracted by solid-phase microextraction (SPME), semi permeable membrane devices (SPMDs), leaching with various solvent mixtures, leaching using additives, and sequential leaching. Bioconcentration factors of PAHs in the earthworms based on equilibrium partitioning theory resulted in poor correlations to observed values. This was most notable for PAHs with high concentrations in the studied soil. Evaluation by principal component analysis (PCA) showed distinct differences between the evaluated techniques and, generally, there were larger proportions of carcinogenic PAHs (4-6 fused rings) in the earthworms. These results suggest that it may be difficult to develop a chemical method that is capable of mimicking biological uptake, and thus estimating the bioavailability of PAHs. PMID- 16713050 TI - Organohalogen contamination in breeding glaucous gulls from the Norwegian Arctic: associations with basal metabolism and circulating thyroid hormones. AB - Exposure to organohalogens in endotherms has been suggested to impose chemically induced stress by affecting functions related to maintenance energy requirements. Effects on basal metabolic rate (BMR) have been suggested to be, in part, mediated through interactions with the thyroid hormones (THs). We investigated the relationships between plasma concentrations of major organochlorines, PBDEs, hydroxylated (OH)- and methoxylated (MeO)-PBDEs and OH-PCBs, circulating TH levels and BMR in breeding glaucous gulls (Larus hyperboreus) from the Norwegian Arctic. Negative associations were found between BMR and concentrations of sigma PCB, Sigma DDT and particularly Sigma chlordane, which combined made up 91% of the total contaminant burden. Levels of THs (thyroxine and triiodothyronine) were not associated significantly with variation of BMR or concentrations of any of the compounds determined. The present study suggests that BMR may be altered in glaucous gulls exposed to high loadings of persistent contaminants in the Norwegian Arctic environment. PMID- 16713051 TI - Self-injurious behavior in human and non-human primates. AB - A review of the literature on human and non-human animal self-injury reveals that there has been little cross-fertilization of ideas between these two domains. Each body of research offers particular strengths, which, if combined, may more keenly inform clinicians and researchers alike about the nature of human self injury. While research on self-injury among humans has necessarily relied upon correlational designs, researchers who study self-injury among non-human primates have more tightly controlled experimental methods at their disposal. Experimental research allows researchers and clinicians to address issues of causality in ways that are not possible with correlational designs. Despite the difficulties of drawing inferences from different species, a review of the experimental data on non-human primate self-injury may give researchers fresh insights into the elusive nature of human self-injury. This article highlights the research and theoretical material pertaining self-injury and identifies ways in which animal models of self-injury might inform research and clinical understanding of human self-injury. PMID- 16713052 TI - Potency and vulnerability: troubled 'selves' in the context of antiretroviral therapy. AB - The concepts of health and self have become intimately entangled in contemporary western society. Health is figured as a marker of identity, as a vehicle of self production and self-actualisation, while the individual is also made increasingly responsible for his or her health. In this paper, we explore how "self" is constituted in discourses that shape the ways in which people understand and do health and medicine, particularly discourses of neoliberalism and of the immune system. Of interest here is how the productive and unpredictable intra-action of medicine and bodies may come to trouble these discursive selves. We situate our discussion in the context of efavirenz, an antiretroviral drug prescribed and consumed for the treatment of HIV infection. This drug, commonly described as "potent", can have a number of troubling effects on a person's everyday sense of self, including insomnia, confusion, cognitive disorders, depression, depersonalisation, psychosis, and suicidal ideation. While efavirenz may be clinically effective in its capacity to suppress the virus, these effects are at odds with the implicit aim of HIV medicine to restore and secure the self by way of immunological integrity and strength. These effects also bring into focus the predicament of choice under the contemporary political conditions of neoliberalism with its emphasis on health as an enterprise of the autonomous, rational self. In exploring first-person accounts, the paper unpacks a number of the binary concepts on which contemporary discourses of health and medicine rely, such as immunity and vulnerability, potency and fragility, rationality and madness, self and non-self, and asks whether the individual under neoliberalism is being asked the impossible. PMID- 16713053 TI - Local understandings of, and responses to, HIV: rural-urban migrants in Tanzania. AB - Migration is an important process of change for rural populations in developing countries. Migration is a primary cause of behaviour change-by their very act of migrating, migrants are different from those who do not migrate. The focus of the current study is male rural-urban migration in Tanzania and its interaction with sexual behaviour. The analysis presents results from a comparison with individual level analyses of two populations, one (composed of recent rural-urban migrants) in an urban area and one made up of residents in a rural area. Detailed migration histories (n=96 rural-urban migrants) and in-depth interviews form the basis of the analysis. Three key research questions are addressed: How does the sexual behaviour of migrants differ from that of rural residents? How do HIV knowledge levels vary between rural-urban migrants and rural residents? What factors are associated with either intentions of behaviour change or reported behaviour? The results are counter-intuitive: rural-urban migrants-both married and unmarried are not having sex in town. Despite limited understanding of the nature of HIV, the migrant population studied here regulates its behaviour in a way that reflects local understandings of the disease. This finding is important, not least because it challenges the view that HIV in sub-Saharan Africa is largely transmitted to rural areas by return migrants. Maasai rural-urban migrants in Tanzania-both married and unmarried-are not having sex in town. The policy and service provision implications of the results are explored. PMID- 16713054 TI - Effects of activation and blockade of 5-HT2A/2C receptors in the dorsal raphe nucleus on sleep and waking in the rat. AB - The effects of the 5-HT(2A/2C) receptor agonist DOI and of the selective 5-HT(2A) or 5-HT(2C) receptor antagonists EMD 281014 and SB-243213, respectively, on spontaneous sleep were studied in adult rats implanted for chronic sleep recordings. The serotonergic ligands were microinjected directly into the dorsal raphe nucleus (DRN). Infusion of DOI (1.4-5.6 mmol) into the DRN induced a significant reduction of REM sleep (REMS) and of the number of REM periods. Following the microinjection of EMD 281014 (5.6 mmol) or SB-243213 (1.4-2.8 mmol) light sleep (LS) was slightly but significantly augmented. Pretreatment with EMD 281014 or SB-243213 antagonized the DOI-induced decrease of REMS. It is proposed that suppression of REMS after DOI microinjection into the DRN is related to the activation of GABAergic projection neurons that synapse cholinergic neurons in the laterodorsal and peduncunculopontine tegmental nuclei (LDT/PPT) involved in the promotion of REMS. PMID- 16713055 TI - Inducible nitric oxide synthase catalyzes ethanol oxidation to alpha-hydroxyethyl radical and acetaldehyde. AB - The physiologic function of nitric oxide synthases, independent of the isozyme, is well established, metabolizing L-arginine to L-citrulline and nitric oxide (NO). This enzyme can also transfer electrons to O2, affording superoxide (O2*-) and hydrogen peroxide (H2O2). We have demonstrated that NOS1, in the presence of L-arginine, can biotransform ethanol (EtOH) to alpha-hydroxyethyl radical (CH3*CHOH). We now report that a competent NOS2 with l-arginine can, like NOS1, oxidize EtOH to CH3*CHOH. Once this free radical is formed, it is metabolized to acetaldehyde as shown by LC-ESI-MS/MS and HPLC analysis. These observations suggest that NOS2 can behave similarly to cytochrome P-450 in the catalysis of acetaldehyde formation from ethanol via the generation of alpha-hydroxyethyl radical when L-arginine is present. PMID- 16713056 TI - Evaluating the genotoxic effects of workers exposed to lead using micronucleus assay, comet assay and TCR gene mutation test. AB - To evaluate the genotoxic effects of lead (Pb) exposure, 25 workers in a workplace producing storage battery were monitored for three genetic end-points using micronucleus (MN) assay, comet assay and TCR gene mutation test. Twenty five controls were matched with workers according to age, gender and smoking. The air Pb concentration in the workplace was 1.26 mg/m(3). All subjects were measured for Pb concentration of blood by atom absorption spectrophotometry. The mean Pb concentration of blood in workers (0.32 mg/l) was significantly higher than that in controls (0.02 mg/l). The results of MN test showed that the mean micronuclei rate (MNR) and mean micronucleated cells rate (MCR) in workers were 9.04+/-1.51 per thousand and 7.76+/-1.23 per thousand, respectively, which were significantly higher than those (2.36+/-0.42 per thousand and 1.92+/-0.31 per thousand) in controls (P<0.01). It was found in the comet assay that the mean tail length (MTL) of 25 workers and 25 controls were 2.42+/-0.09 and 1.02+/-0.08 microm, respectively, there was significant difference between workers and controls for MTL (P<0.01), also the difference of the mean tail moment (MTM) between workers (0.85+/-0.05) and controls (0.30+/-0.09) was very significant (P<0.01). However, in TCR gene mutation assay Mfs-TCR of workers and controls were 1.69+/-0.15 x 10(-4) and 1.74+/-0.17 x 10(-4), respectively, there was no significant difference between workers and controls (P>0.05). The results of our study indicated that the genetic damage was detectable in 25 workers occupationally exposed to lead. PMID- 16713057 TI - Psychrophilic superoxide dismutase from Pseudoalteromonas haloplanktis: biochemical characterization and identification of a highly reactive cysteine residue. AB - A psychrophilic superoxide dismutase (SOD) has been characterized from the Antarctic eubacterium Pseudoalteromonas haloplanktis (Ph). PhSOD is a homodimeric iron-containing enzyme and displays a high specific activity, even at low temperature. The enzyme is inhibited by sodium azide and inactivated by hydrogen peroxide; it is also very sensitive to peroxynitrite, a physiological inactivator of the human mitochondrial Mn-SOD. Even though PhSOD is isolated from a cold adapted micro-organism, its heat stability is well above the maximum growth temperature of P. haloplanktis, a feature common to other Fe- and Mn-SODs. The primary structure of PhSOD was determined by a combination of mass spectrometry and automated Edman degradation. The polypeptide chain is made of 192 amino acid residues, corresponding to a molecular mass of 21251 Da. The alignment with other Fe- and Mn-SODs showed a high amino acid identity with Fe-SOD from Vibrio cholerae (79%) and Escherichia coli (70%). A significant similarity is also shared with human mitochondrial Mn-SOD. PhSOD has the unique and highly reactive Cys57 residue, located in a variable region of the protein. The three-dimensional model of the PhSOD monomer indicates that Cys57 is included in a region, whose structural organization apparently discriminates between dimeric and tetrameric SODs. This residue forms a disulfide adduct with beta-mercaptoethanol, when this reducing agent is added in the purification procedure. The reactivity of Cys57 leads also to the formation of a disulfide bridge between two PhSOD subunits in specific denaturing conditions. The possible modification of Cys57 by physiological thiols, eventually regulating the PhSOD functioning, is discussed. PMID- 16713059 TI - The effect of glucose administration on the emotional enhancement effect in recognition memory. AB - Previous research has demonstrated that glucose administration improves memory performance. However few studies have addressed the effects of glucose on emotional material that by nature already enjoys a memory advantage. The aim of the present research was therefore to investigate whether the memory facilitation effect associated with glucose would emerge for emotional words. Experiment 1 demonstrated that negative words were better recognized and remembered than positive and neutral words. Experiment 2 further explored these effects under conditions of glucose administration and an aspartame control. The results revealed that both the aspartame and glucose groups replicated the results from Experiment 1. The present research therefore demonstrated that the glucose facilitation effect did not emerge for material that already benefits from a memory advantage. These results also raise the question of whether the dose response relationship previously associated with glucose administration is applicable when the information being processed is of an emotional nature. PMID- 16713060 TI - Reasons for early interventions by gynaecologists in a clinical follow-up study on cervical intraepithelial neoplasia. AB - OBJECTIVE: In a previous clinical follow-up study on psychosocial factors and cervical intraepithelial neoplasia (CIN), we had noticed a high percentage of early interventions to the cervix by gynaecologists, in discordance with the research protocol. The aim of the current study was to investigate the reasons for these interventions. STUDY DESIGN: We retrospectively analysed the study records of the clinical study, concerning the follow-up of 98 patients with a CIN 1 or 2 lesion for 2.25 years by colposcopy and cervical smears. We recorded the number of early interventions that had occurred in the clinical study as well as the reasons for these interventions. A distinction was made between acceptable and unacceptable reasons for early intervention. RESULTS: During follow-up, 39 out of 98 women (40%) had undergone intervention in discordance with the protocol. No acceptable reason for these interventions was found for 67% of these patients (26/39). In the majority of these cases either no reason for the intervention had been given, or the gynaecologist had feared for progression of the CIN lesion. CONCLUSION: Gynaecologists tend to intervene during follow-up of CIN, partly out of fear for progression. This tendency may complicate follow-up studies on CIN. PMID- 16713061 TI - Effect of vaginal delivery on endosonographic anal sphincter morphology. AB - OBJECTIVE: To describe the effect of vaginal delivery with no clinically recognized sphincter tear on endosonographic anal sphincter morphology and sphincter pressure and to relate endosonographic results to anal sphincter pressure and anal incontinence score. STUDY DESIGN: Thirty-two nullipara underwent anal endosonography and anal manometry in the third trimester of pregnancy, 2 weeks and 6 months post-partum. The sphincter defect scores (1-16) and the thickness and length of the sphincters were measured by endosonography, and sphincter pressures and manometric sphincter lengths were determined. The Wexner incontinence score (1-20) was used to classify anal incontinence 6 months post-partum. RESULTS: Five (16%) women had small endosonographic anal sphincter defects (score 3-4) before delivery. Eight women (25%; confidence interval 11 43%) had new defects detected post-partum, five small, one moderate (score 7), and two large (score 10-11). Six (75%) of eight women with new defects post partum had undergone episiotomy versus five (21%) of 24 women with no new defects (p = 0.02). Six months after delivery 16 (50%) women reported anal incontinence, and there was a positive correlation between the endosonographic defect score 6 months post-partum and the Wexner incontinence score. The sphincter was significantly longer during pregnancy than 6 months post-partum. CONCLUSION: New sphincter defects may arise after vaginal delivery without any clinically recognizable sphincter tear. There is a positive correlation between the endosonographic defect score 6 months post-partum and the Wexner incontinence score. PMID- 16713062 TI - Management of multicentric lesions of the lower genital tract. AB - OBJECTIVES: To report management and outcome of multicentric lesions of the lower genital tract. To define risk factors of recurrence. STUDY DESIGN: Retrospective review of multicentric dysplasias treated in our colposcopic clinic between 1996 and 2003. Multicentric dysplasias included CIN with VAIN and/or VIN. After primary treatment, follow-up was colposcopic, cytologic and virologic. RESULTS: Forty-four patients presented multicentric lesions out of 998 patients referred for CIN (4.4%). The average age was 36.8 years. Immunologic disorders were present in 20.4%. Ninety-one percent had cervicovaginal or cervicovulvar lesions, only 9% had three sites of genital dysplasia. 53.3% of lesions were concomitant. 79.5% of CIN were high grade, 62.5% of VAIN low grade and 62.5% of VIN high grade. Therapeutic modalities were as follows: conization for CIN (70.4%), CO2 laser for VAIN (33.3%) and surgery for VIN (41.7%). Forty patients were followed and had at least one post-treatment cytologic control; 55% of them had residual disease. Out of the 23 patients with at least two negative controls after treatment, 43.5% presented recurrence. Risk of recurrence was not statistically bound to such parameters as tabagism, immunologic disorder, high grade lesions, non-surgical treatment, and persistence of HPV infection after treatment. CONCLUSION: Multicentric dysplasias are associated with high rate of residual lesion and recurrence. Management of these lesions require long term follow-up. PMID- 16713063 TI - Elimination of the C-cap in ubiquitin - structure, dynamics and thermodynamic consequences. AB - Single amino acid substitutions rarely produce substantial changes in protein structure. Here we show that substitution of the C-cap residue in the alpha-helix of ubiquitin with proline (34P variant) leads to dramatic structural changes. The resulting conformational perturbation extends over the last two turns of the alpha-helix and leads to enhanced flexibility for residues 27-37. Thermodynamic analysis of this ubiquitin variant using differential scanning calorimetry reveals that the thermal unfolding transition remains highly cooperative, exhibiting two-state behavior. Similarities with the wild type in the thermodynamic parameters (heat capacity change upon unfolding and m-value) of unfolding monitored by DSC and chemical denaturation suggests that the 34P variant has comparable buried surface area. The hydrophobic core of 34P variant is not packed as well as that of the wild type protein as manifested by a lower enthalpy of unfolding. The increased mobility of the polypeptide chain of this ubiquitin variant allows the transient opening of the hydrophobic core as evidenced by ANS binding. Taken together, these results suggest exceptional robustness of cooperativity in protein structures. PMID- 16713064 TI - The potential of cost-effective coconut husk for the removal of toxic metal ions for environmental protection. AB - Coconut (Cocos nucifera) husk, an agricultural waste, has been thoroughly investigated for the removal of toxic Cd(II), Cr(III) and Hg(II) ions from aqueous media. The parameters like nature and composition of electrolyte, concentration of toxic ions, dosage of coconut husk, and equilibration time between the two phases were optimized for their maximum accumulation onto the solid surface. The effect of common ions on the uptake of metal ions has been monitored under optimal conditions. The variation of retention of each metal ion with temperature was used to compute the thermodynamic quantities DeltaH, DeltaS and DeltaG. The values 18.1+/-0.6 kJmol(-1), 74+/-2 Jmol(-1)K(-1), and -3.8+/ 0.04 kJmol(-1) at 298 K; 10.8+/-0.8 kJmol(-1), 48.8+/-2.7 Jmol(-1)K(-1), and 4.6+/-0.3 kJmol(-1) at 298 K; and -37.4+/-2k Jmol(-1), 105+/-7 Jmol(-1)K(-1) and 2.58+/-0.5 kJmol(-1) at 298 K were obtained for Cd(II), Cr(III) and Hg(II) ions, respectively. The sorption data were analysed by applying different sorption isotherms. The sorption capacity and energy were evaluated for each metal ion. The values of the Freundlich constants 1/n and C(m) were 0.92+/-0.04 and 52.6+/ 22.2 mmolg(-1); 0.85+/-0.05 and 56.0+/-0.03 mmolg(-1); and 0.88+/-0.03 and 6.84+/ 0.45 mmolg(-1) for Cd(II) Cr(III) and Hg(II) ions, respectively. Similarly, the Dubinin-Radushkevich (D-R) constants beta, X(m,) and E were evaluated for the three metal ions. To check the selectivity of the sorbent, sorption of a number of elements was measured under similar conditions. Separation of Zn(II) from Cd(II); Cr(III) from I(I), Zr(IV), Se(IV), and Hg(II) from Se(IV) and Zn(II) can be achieved using this sorbent. This cheap material has potential applications in analytical chemistry, water decontamination, industrial effluent treatment and in pollution abatement. PMID- 16713065 TI - Management and survival of screen-detected prostate cancer patients who might have been suitable for active surveillance. AB - OBJECTIVE: Screening practices for prostate cancer have resulted in an increasing incidence of prostate cancers. Our knowledge about which prostate cancers are life threatening and which are not is limited. Thus, for ethical, medical, and economic reasons we need to define which patients can be managed by active surveillance. METHODS: From 1993 through 1999, men from the Rotterdam section of the European Randomized study of Screening for Prostate Cancer (ERSPC) were screened by two strict protocols, which were based on prostate-specific antigen (PSA), digital rectal examination, and transrectal ultrasound. For this study, men with criteria that reflect current active surveillance studies were selected: those with a biopsy Gleason score < or =3+3 in two or fewer cores, with a PSA density <0.2 and a maximum PSA-level of 15 ng/ml. Clinical stage had to be T1C or T2. RESULTS: Of the 1,014 prostate cancers detected in the prevalence screen, 293 men (28.9%) met the criteria for active surveillance. Their mean age was 65.7 and the mean PSA level was 4.8 ng/ml. Radical prostatectomy was elected by 136 men (46.4%), radiotherapy by 91 (31.1%), and watchful waiting by 64 (21.8%). The mean follow-up was 80.8 months. The eight-year prostate cancer-specific survival was 99.2%; the overall survival was 85.4%. Nineteen men who chose watchful waiting changed to definitive treatment during follow-up. CONCLUSION: Only three men died of prostate cancer, none of whom were on watchful waiting. Our observations provide preliminary validation of the arbitrary selection criteria for active surveillance. PMID- 16713066 TI - Five-year outcomes of the tension-free vaginal tape procedure for treatment of female stress urinary incontinence. AB - OBJECTIVES: We evaluated the long-term efficacy and safety of a tension-free vaginal tape (TVT) procedure for the treatment of female stress urinary incontinence (SUI) in a Korean population. METHODS: We included 134 patients (mean age, 52.3+/-9.3 yr) who underwent the TVT procedure for SUI in three institutions and followed for 5 yr (mean, 67.0 mo; range, 60-76 mo) postoperatively. We analysed voiding diaries and complete multichannel urodynamic studies preoperatively as well as cough stress tests, uroflowmetry, and questionnaires postoperatively. RESULTS: The overall 5-yr success rates (cure/improved) were 94.9% (76.9% and 18.0%, respectively), with an 86.6% patient satisfaction rate. Although the success rates between 1 and 5 yr were similar (97.7% vs. 94.9%), the cure rate decreased from 90.1% to 76.9% (p<0.001) at 5 yr. The 5-yr cure rate for mixed urinary incontinence (MUI) was 72.0%, which was not significantly different from pure SUI (78.0%, p>0.05). Maximal flow rate dropped from 25.9+/-10.3ml/s to 20.4+/-8.6ml/s at 1 mo postoperatively and recovered to 24.8+/-8.5ml/s at 5 yr. Complications included bladder perforation in 5 patients (3.7%), tape cutting or release in 11 (8.2%), and persistent suprapubic pain in 3 (2.2%). Urgency and urge incontinence improved in 46.7% and 48.0% of patients, respectively. CONCLUSIONS: TVT was an effective and safe procedure for SUI and MUI with high success rates in the long-term follow-up. It also improved concomitant overactive bladder symptoms and initially reduced postoperative urine flow, which recovered over time. PMID- 16713068 TI - Dendritic cells for the treatment of metastatic renal cell carcinoma: at a low ebb? PMID- 16713067 TI - Italian validation of the urogenital distress inventory and its application in LUTS patients. AB - OBJECTIVES: The objective of this study was to validate the Italian version of the Urogenital Distress Inventory (UDI) in a sample of women with lower urinary tract symptoms (LUTS). METHODS: The linguistic validation of the questionnaire was performed through a multistep process: backward and forward translations coordinated by clinical investigators, followed by a pretest. The final version was administered to a larger sample of female patients, aged 18 years or older who had been having LUTS for at least 3 months, numbering 53 subjects. To evaluate test-retest reliability, patients were re-rated after 1 week. To test the questionnaire's capacity to discriminate women with or without LUTS (cases and controls, respectively), a sample of 53 healthy women was enrolled. A 72-h voiding diary was used as a gold standard and compared with the UDI. RESULTS: The correlation coefficient between ratings was >or=0.80, and the discriminant power between cases and controls was confirmed. The UDI showed good internal consistency for all domains, except irritative symptoms (total score's Cronbach alpha=0.86). Factor analytic structure revealed urinary incontinence to be opposite to the other urologic symptoms, with bed wetting being loaded separately. The average daily number of urgent micturitions was higher in patients who reported they "experience a strong feeling of urgency to empty bladder" in the UDI than those ones who did not (p<0.01). CONCLUSIONS: The Italian version of the UDI is a valid and robust instrument, which can now be used reliably in daily practice and clinical research. PMID- 16713069 TI - Interview with Jack Schalken. PCA3 and its use as a diagnostic test in prostate cancer. Interview by Christine McKillop. PMID- 16713070 TI - Holmium laser enucleation of the prostate versus open prostatectomy for prostates >70 g: 24-month follow-up. AB - OBJECTIVE: Prospectively evaluate perioperative outcomes and 2-yr follow-up after holmium laser enucleation (HoLEP) and standard open prostatectomy (OP) for treating benign prostatic hyperplasia-related obstructed voiding symptoms, with prostates >70 g. METHODS: From March 2003 to December 2004, 80 consecutive patients were randomised for surgical treatment with HoLEP (n=41) or standard OP (n=39). All patients were preoperatively assessed with International Prostate Symptom Score and International Index of Erectile Function questionnaires and complete urodynamic evaluation. Intraoperative and perioperative parameters such as blood loss, catheter removal, and hospital stays were assessed. Early and late complications were recorded. Patients were evaluated at 1-, 3-, 12-, and 24-mo follow-ups with the same tests. RESULTS: Operating room time was significantly shorter for the OP group (72.09+/-21.22 min vs. 58.31+/-11.95 min, p<0.0001); catheter removal (1.5+/-1.07 d and 4.1+/-0.5 d, p<0.001) and hospital stay (2.7+/ 1.1 d vs. 5.4+/-1.05 d, p<0.001) were shorter in the HoLEP group. Blood loss was less and blood transfusions fewer in the HoLEP group (p<0.001). In both groups urodynamic and uroflowmetry findings improved from baseline, were still evident at the 24-mo follow-up, and were comparable between the two groups. Late complications were also comparable. CONCLUSIONS: HoLEP is a feasible technique for treating large prostates. Functional results are similar to OP at the 2-yr follow-up. Reduced catheterisation, hospital stay, and blood loss make HoLEP an attractive option for the treatment of large prostates. PMID- 16713071 TI - Changing times: the evolution of puberty. AB - An evolutionary and life history perspective is used to consider the evolution of puberty. The age of menarche would have evolved by the Neolithic to be matched to social maturity. It is suggested that in developed countries menarche is now returning to a similar age as in the Neolithic as infection and undernutrition, features of post-Neolithic society, have reduced impact. But recently the psychosocial expectations on adolescents in western societies have changed and social maturity now significantly follows menarche. The implications of the developing mismatch between the ages of biological puberty and social maturation are discussed. Evolutionary arguments are presented to explain the unique pubertal growth spurt of humans. Moreover, a life history perspective can reconcile the apparently conflicting observations that both poor fetal growth and better childhood nutrition are associated with earlier menarche. PMID- 16713073 TI - Loss of promyelocytic leukemia protein in human gastric cancers. AB - To clarify the clinical implications of promyelocytic leukemia (PML) expression in gastric carcinomas, the expression of PML was analyzed in large series of gastric carcinoma by immunohistochemistry, western blotting and reverse transcription-PCR. PML protein expression was reduced or abolished in gastric carcinomas (31.7 and 10.6%, respectively) by immunohistochemistry. PML protein loss was associated with more lymphatic invasion, higher pTNM stage, and worse patient survival. Only one gastric carcinoma cell line showed loss of PML, and the PML protein re-appeared after the treatment of proteasome inhibitor in this cell line. We conclude that PML protein loss occurs in a minority of gastric carcinomas during carcinogenesis and progression, and suggest the proteasome dependent pathway as a mechanism of PML protein loss. PMID- 16713072 TI - Encephaloradiculomyelitis associated to HHV-7 and CMV co-infection in immunocompetent host. AB - An active co-infection with CMV and HHV-7 has been never described in immunocompetent patients. The authors describe a case of encephaloradiculomyelitis in an immunocompetent man. Polymerase chain reaction (PCR) performed on cerebrospinal fluid (CSF) showed positivity for DNA of Cytomegalovirus (CMV) and Herpes-virus type 7 (HHV-7), whereas the same test applied on peripheral blood mononuclear cells gave negative result. These results are highly supportive of an infection of the central and peripheral nervous systems, caused by CMV and HHV7. Such viral co-infection has only been described in immune-depressed patients with CMV disease, in which HHV-7 was supposed to act as a cofactor, enhancing clinical manifestations. The same mechanism is presumably responsible for the development of encephaloradiculomyelitis clinical signs in the present case. This is the second case in which DNA of HHV-7 has been found in the CSF of an adult immunocompetent patient. This novel observation suggests that the search for viral DNA in the CSF should be performed also in immunocompetent patients. PMID- 16713074 TI - Arsenic inhibits induction of cytochrome P450 1A1 by 2,3,7,8-tetrachlorodibenzo-p dioxin in human hepatoma cells. AB - The aim of this study was to examine the arsenic effect on activation of aryl hydrocarbon receptor (AhR)-mediated gene expression by 2,3,7,8-tetrachlorodibenzo p-dioxin (2,3,7,8-TCDD) in human hepatoma cells. The human hepatoma Huh7 cells were treated with sodium arsenite (NaAsO2) from 0.5 to 20 microM for 24 h. Our data revealed that NaAsO2 < or = 10 microM caused no significant cytotoxic effect on Huh7 cells (p>0.05). We also established a dioxin-responsive element (DRE) mediated Chemical Activated LUciferase eXpression (CALUX) cell line, Huh7-DRE Luc, by stable transfection of Huh7 with a DRE-driven firefly luciferase reporter plasmid (4xDRE-TATA-Luc). Treatments of Huh7-DRE-Luc and Huh7 with NaAsO2 attenuated the 2,3,7,8-TCDD-induced DRE-CALUX and cytochrome P450 1A1 (CYP1A1) activations, respectively, in a dose-dependent manner. We found that the calculated CALUX-toxic equivalent (TEQ) levels induced by cotreatment of NaAsO2 > or = 3.0 microM and 10 nM 2,3,7,8-TCDD were significantly lower than that induced by 2,3,7,8-TCDD alone (p<0.05). In the present study, we demonstrated that arsenic not only inhibited the TCDD-induced CYP1A1 activation but also interfered with DRE-CALUX bioassay in human hepatoma cells. Our finding also suggests that extensive cleanup of sample for removal of any possible interfering factor is critical to guarantee the accuracy of dioxin-TEQ levels using DRE-CALUX bioassay. PMID- 16713075 TI - Mechanochemical decomposition of PVC by using La2O3 as additive. AB - A polyvinyl chloride (PVC, [CH2CHCl]n) sample was ground with one of the following oxide samples, lanthanum oxide (La2O3) and calcium oxide (CaO), as a reference, to investigate its mechanochemical (MC) decomposition. According to the XRD patterns for the ground products, peaks of lanthanum oxy-chloride, LaOCl which is water insoluble, were observed to appear in the patterns of the mixture samples at a molar ratio of (PVC: La2O3 = 1:2) and (1:4), after 20 min grinding, while the patterns of the ground PVC-CaO mixtures show the formation of CaOHCl, which is water soluble, in the products after 2 h grinding. TG patterns taken for the PVC-La2O3 mixtures ground for different periods of time do not indicate significant weight loss in the range from 200 to 300 degrees C, and this suggests that La2O3 is more effective additive than CaO in the MC decomposition for PVC. FT-IR patterns for organic phases in the ground mixture show the reduction of CCl bindings in the PVC structure. The MC reaction between PVC and La2O3 is accompanied by the formation of C-O single bonds, to form LaOCl, which is insoluble in water at ambient condition. This is the reason why the yield of Cl extracted by the water leaching reaches only less than 50%, while it reaches almost 100% in the yield of Cl from the PVC-CaO mixture ground for 2 h or more. PMID- 16713076 TI - Diaion SP-850 resin as a new solid phase extractor for preconcentration separation of trace metal ions in environmental samples. AB - A solid phase extraction method was developed for the preconcentration and separation of trace amounts of chromium, manganese, iron, cobalt, copper, cadmium and lead from environmental samples by complexation with alpha-benzoin oxime followed by adsorption onto Diaion SP-850-solid phase extraction column. One molar per liter HNO(3) was used as eluent. The recoveries of analytes at pH 8.0 with 700 mg of resin were greater than 95% without interference from alkaline, earth alkaline and some metal ions. The detection limits by three sigma for analyte ions were 0.65 microg l(-1) for Cr(III), 0.42 microg l(-1) for Mn(II), 0.28 microg l(-1) for Fe(III), 0.73 microg l(-1) for Co(II), 0.30 microg l(-1) for Cu(II), 0.47 microg l(-1) for Cd(II) and 0.50 microg l(-1) for Pb(II). The validation of the procedure was performed by the analysis of the certified standard reference materials. The presented procedure was applied to the determination of analytes in tap, river and sea waters, rice, wheat, canned tomato and coal samples with successfully results (recoveries greater than 95%, R.S.D.'s lower than 8%). PMID- 16713077 TI - An investigation of heavy metal biosorption in relation to C/N ratio of activated sludge. AB - The effect of C/N ratio of activated sludge on heavy metal biosorption was investigated. Three sets of semi-continuous reactors with different feed C/N ratios (9, 21 and 43 mg COD/mg TKN) were set up. Sorption equilibrium tests have indicated that the biosorptive capacity of activated sludge was highly dependent on metal species and the C/N ratio. The increase in C/N ratio resulted in an increase in the Cd(II) sorption capacity of activated sludge whereas it decreased the Cu(II) sorption capacity. As for Zn(II), a different behavior was observed such that, the highest and lowest capacities have occurred at C/N ratio of 21 and 43, respectively. For Ni(II) biosorption, isotherm tests produced greatly scattered data; so, it was not possible to obtain any plausible result to indicate the relationship between maximum adsorptive capacity and C/N ratio. The accompanying release of Ca(II) and Mg(II) ions and also carbohydrates into the solution during biosorption have indicated that ion exchange mechanism was involved however, was not the only mechanism during the sorption process. PMID- 16713078 TI - Adsorption of copper and lead in aqueous solution onto bentonite modified by 4' methylbenzo-15-crown-5. AB - The adsorption of heavy metal ions (Cu2+ and Pb2+) onto organobentonite modified by 4'-methylbenzo-15-crown-5 (MB15C5) from bentonite compared with natural bentonite (N-Bentonite) is described in this paper. The kinetic parameters of the models are calculated and discussed, and closely fitted a pseudo-second-order model in all cases. For bentonite modified with MB15C5 (MB15C5-Bentonite) and N Bentonite, the equilibrium data closely fitted the Langmuir model and showed the following affinity order: Pb2+ > Cu2+, and the adsorption capacity of MB15C5 Bentonite is higher than that of N-Bentonite for Pb2+ and Cu2+. The effect of pH is examined over the range 1.5-6. The adsorption of Cu2+ and Pb2+ increases with increasing pH and the adsorption of Cu2+ and Pb2+ reaches a maximum at pH 3.5-6. PMID- 16713079 TI - Pentachlorophenol removal from aqueous matrices by sorption with almond shell residues. AB - Sorption with activated carbon has been the technique preferred for pentachlorophenol (PCP) removal from contaminated waters, but regeneration needs and high operation costs are supporting a renewed interest in the search for alternative sorbents. Among them, almond shell, an agricultural by-product, provides interesting economical advantages, once shells account for 50% (in mass) of the whole almond. In this work, the capacity of almond shells to remove PCP from waters without previous activation was studied in batch conditions. While PCP analysis was performed solid-phase microextraction (SPME) followed by gas chromatography with electron capture detection (GC-ECD), mercury porosimetry and Fourier transform infrared spectroscopy (FTIR) provided a preliminary physical and chemical characterization of the sorbent. Almond shells were essentially a macroporous material, with an average surface area of 12.9+/-2.8 m2/g. The efficiency of PCP removal was 93+/-14%, in 24 h, with an initial concentration of 100 microg/l PCP and 5 microg PCP/g shell. Isotherm data adjusted better to Freundlich equation, where K(F) and 1/n were 0.075+/-0.081 mg(1-1/n) l(1/n) and 1.882+/-0.289, respectively. Average desorption efficiency was 7%, indicating strong adsorption capacity. Results proved that almond shells may be an excellent low-cost alternative for PCP removal from contaminated waters. PMID- 16713080 TI - Indicating the parameters of the state of degradation of municipal solid waste. AB - This study related to the analysis of the physical (granulometry and composition) and chemical (organic matter, organic carbon and nitrogen contents) characteristics, as well as those relating to release (leaching tests and determining the methanogene potential) of several domestic wastes, with an aim of evaluating stabilization indicators. Values thresholds for a known stabilized waste were thus deduced by correlation (% paper-cardboard = 0-1; % volatile solid (VS) = 18-19; % OC = 5-6; % fines = 44-45; % degraded components = 75-76, COD of leachate = 141-155 mg O2/L; DOC from leachate = 45-49 mg C/L and 0.9-1 m3 CH4/t of dry waste). However, these values thresholds remain specific to the method employed for the analyses. The results obtained represent a considerable advance in the definition of a waste stabilized state and propose the importance of certain parameters, such as the paper-cardboard content and the measurement of leachates by using the SUVA index for determining a stabilization state. PMID- 16713081 TI - Electrokinetic movement of hexachlorobenzene in clayed soils enhanced by Tween 80 and beta-cyclodextrin. AB - This study describes the comparative behavior of hexachlorobenzene (HCB) contaminated clayed soils in an electrokinetic (EK) system enhanced by Tween 80 and beta-cyclodextrin (beta-CD). The pH of the soils was controlled by Na2CO3/NaHCO3 buffer. Negligible HCB movement was observed when NaOH or Na2CO3/NaHCO3 buffer was used as anodic flushing solution. While Tween 80 or beta CD was introduced to Na2CO3/NaHCO3 buffer, obvious HCB movement was achieved. Although beta-CD led to a less desorption of HCB from kaolin than Tween 80, the removal of HCB with beta-CD was much higher than that with Tween 80 in the EK system. Tween 80 could be sorped by kaolin more than beta-CD, which was responsible for the result. The mechanism of the movement of HCB was proposed as the enhanced desorption of HCB from soil, the dissolving of HCB in the soil pore fluid and the movement of HCB with the electroosmotic flow. Obvious movement of HCB was also observed in the EK treatment of real HCB-contaminated clayed soil enhanced by beta-CD. It is an alternative approach to use facilitating agents such as beta-CD to enhance the EK movement of HCB in the contaminated clayed soils. PMID- 16713082 TI - Mechanisms of the removal of hexavalent chromium by biomaterials or biomaterial based activated carbons. AB - Three papers published during recent 2 years in Journal of Hazardous Materials made a mistake in analyzing chromium species in aqueous solution, resulting in incorrect elucidation of Cr(VI) biosorption; the Cr(VI) was removed from aqueous solution systems by 'anionic adsorption'. However, it has been proved that Cr(VI) is easily reduced to Cr(III) by contact with organic materials under acidic conditions because of its high redox potential value (above +1.3 V at standard condition). Therefore, it is strongly possible that the mechanism of Cr(VI) removal by biomaterials or biomaterial-based activated carbons is not "anionic adsorption" but "adsorption-coupled reduction". Thus, for researches of Cr(VI) biosorption, researchers have to analyze not only Cr(VI) but also total Cr in aqueous solution and to check the oxidation state of chromium bound on the biomaterials or activated carbons. PMID- 16713083 TI - Application of microwave-assisted extraction to the analysis of PCBs and CBzs in fly ash from municipal solid waste incinerators. AB - Polychlorinated biphenyls (PCBs) and chlorobenzenes (CBzs) are two classes of dioxin precursors formed in municipal solid waste incinerators (MSWIs) producing negative health effects similar to those of dioxins. Reducing the analytical time required for determining the concentrations of these compounds in MSWIs is important for quickly evaluating their importance and associated health risks. In the present study, microwave-assisted extraction (MAE) is compared with traditional Soxhlet extraction (SE) to determine the extraction efficiencies attained for PCB and CBz analysis. The efficiencies of MAE are compared with those of SE under various experimental conditions, using fly ash spiked with standards. Water is used as a safe and environmentally friendly solvent in MAE for PCB and CBz analyses and MAE has high extraction efficiency for spiked fly ash compared with that of SE. Furthermore, the extraction time and organic solvent consumption are reduced with MAE compared with SE. The optimum conditions for MAE established in this study are using a 30-ml volume of toluene/acetone (1/1) or a 15-ml volume of toluene, samples with less than 60% water content (WC), and an irradiation time of 15 min. PMID- 16713084 TI - The application of tree bark as bio-indicator for the assessment of Cr(VI) in air pollution. AB - The impact of a chromium smelter on pollution was evaluated by determining Cr(VI) in topsoil, grass and tree bark by electrothermal atomic absorption spectrometry (ETAAS). It was found that bark reflected the levels of air pollution better than soil and grass due to its high accumulative ability of Cr(VI). The tree bark was contaminated with Cr(VI) by a factor of 9 than in soil. It is therefore suggested that the bark be used as an indicator of air pollution for long-term exposure. The concentration of Cr(VI) in the bark was always a fraction of the total concentration of Cr and ranges between 1.6 and 3%. The method used in the preparation of samples was validated by the analysis of certified reference materials. PMID- 16713085 TI - Permeation of Comite through protective gloves. AB - The goal of the study was to assess how protective disposable (Safeskin) and chemical protective (Sol-Vex) nitrile gloves were against Comite emulsifiable concentrate formulation containing propargite (PROP) as active pesticidal ingredient, because there were no explicit recommendations for the gloves that should be worn for hand protection. The glove material was exposed in ASTM-type I PTC-600 permeation cells at 30.0+/-0.5 degrees C, and gas chromatography-mass spectrometry used for PROP analysis. Aqueous solutions of Comite at 40.4 mg/mL permeated both Safeskin and Sol-Vex nitrile by 8h. Safeskin showed a mean PROP mass permeated of 176+/-27 microg after 8h compared with a mean mass permeated for Sol-Vex of 3.17+/-4.08 microg. Thus, Sol-Vex was about 56 times more protective than Safeskin for an 8-h exposure. However, the kinetics of the permeation revealed that Safeskin can be worn for at least 200 min before disposal. When undiluted Comite challenged both types of nitrile, much faster permeation was observed. Safeskin gloves showed two steady state periods. The first had lag times (t(l)) values of about 1h, although normalized breakthrough times (t(b)) were < 10 min. The second steady state rate (P(s)) was on average four times the rate of the first period, and the second steady state period t(l) was about three times as long as that of the first steady state period, and about the same t(l) as for the aqueous solution. Sol-Vex gloves exposed continuously to undiluted Comite permeated above the normalized breakthrough threshold beyond 2.7h. A risk assessment revealed that the PROP skin permeation rate of 7.1 ng cm( 2)h(-1) was much slower than the first steady state Safeskin glove P(s) of 62,000 ng cm(-2)h(-1). Infrared analysis showed that the glove surfaces were not degraded by the Comite challenge. The chemically protective Sol-Vex gloves protected adequately against undiluted formulation for about 2.7h, whereas they provided protection for nearly 8h when the formulation was diluted with water to the highest concentration for field application. In contrast, the disposable Safeskin gloves did not protect at all for the undiluted formulation, but did for 200 min when the formulation was diluted with water to the highest concentration for spraying. PMID- 16713086 TI - Electrochemical degradation of amaranth aqueous solution on ACF. AB - The degradation of Amaranth, a kind of azo dye, has been studied under galvanostatic model with activated carbon fiber (ACF) electrode in aqueous solution with electrochemical method. The ACF was used as anode and cathode, respectively for the decolorization process. The onset oxidation potential and reduction potential for Amaranth on ACF were respectively ascertained at 0.6 and 0.4 V. During the range of -1.1 to 0.50 mA cm(-2), the decolorization was clarified into three processes as electroreduction, adsorption and electrooxidation. There were little contributions to the color and COD removals for the process of adsorption. The color removal can be up to 99% when the current density was 0.50 mA cm(-2). The maximum COD removal was 52% for the process of electrooxidation. Hundred percent color removal was obtained when the current density of -1.0 mA cm(-2) was applied. The maximum COD removal was 62% for the electroreduction. The COD removal results from the adsorption of products for the decolorization process of electrooxidation or electroreduction. PMID- 16713087 TI - Electrochemical treatment of textile dyes and dyehouse effluents. AB - The electrochemical oxidation of textile effluents over a titanium-tantalum platinum-iridium anode was investigated. Batch experiments were conducted in a flow-through electrolytic cell with internal recirculation at current intensities of 5, 10, 14 and 20A, NaCl concentrations of 0.5, 1, 2 and 4% and recirculation rates of 0.81 and 0.65 L/s using a highly colored, synthetic effluent containing 16 textile dyes at a total concentration of 361 mg/L and chemical oxygen demand (COD) of 281 mg/L. Moreover, an actual dyehouse effluent containing residual dyes as well as various inorganic and organic compounds with a COD of 404 mg/L was tested. In most cases, quantitative effluent decolorization was achieved after 10 15 min of treatment and this required low energy consumption; conversely, the extent of mineralization varied between 30 and 90% after 180 min depending on the operating conditions and the type of effluent. In general, treatment performance improved with increasing current intensity and salinity and decreasing solution pH. However, the use of electrolytes not containing chloride (e.g. FeSO4 or Na2SO4) suppressed degradation. Although the acute toxicity of the actual effluent to marine bacteria Vibrio fischeri was weak, it increased sharply following treatment, thus suggesting the formation of persistent toxic by products. PMID- 16713088 TI - Cellular automata-based forecasting of the impact of accidental fire and toxic dispersion in process industries. AB - The strategies to prevent accidents from occurring in a process industry, or to minimize the harm if an accident does take place, always revolve around forecasting the likely accidents and their impacts. Based on the likely frequency and severity of the accidents, resources are committed towards preventing the accidents. Nearly all techniques of ranking hazardous units, be it the hazard and operability studies, fault tree analysis, hazard indice, etc.--qualitative as well as quantitative--depend essentially on the assessment of the likely frequency and the likely harm accidents in different units may cause. This fact makes it exceedingly important that the forecasting the accidents and their likely impact is done as accurately as possible. In the present study we introduce a new approach to accident forecasting based on the discrete modeling paradigm of cellular automata. In this treatment an accident is modeled as a self evolving phenomena, the impact of which is strongly influenced by the size, nature, and position of the environmental components which lie in the vicinity of the accident site. The outward propagation of the mass, energy and momentum from the accident epicenter is modeled as a fast diffusion process occurring in discrete space-time coordinates. The quantum of energy and material that would flow into each discrete space element (cell) due to the accidental release is evaluated and the degree of vulnerability posed to the receptors if present in the cell is measured at the end of each time element. This approach is able to effectively take into account the modifications in the flux of energy and material which occur as a result of the heterogeneous environment prevailing between the accident epicenter and the receptor. Consequently, more realistic accident scenarios are generated than possible with the prevailing techniques. The efficacy of the approach has been illustrated with case studies. PMID- 16713089 TI - Preparation of an aminated macroreticular resin adsorbent and its adsorption of p nitrophenol from water. AB - A chloromethylated styrene-divinylbenzene copolymer was post-cross-linked through Friedel-Crafts reaction to get a macroreticular resin adsorbent NA-01, and then chemically modified by dimethylamine to obtain an aminated adsorbent NA-01A. Batch adsorption runs of p-nitrophenol from aqueous solution onto adsorbent NA 01A were conducted to evaluate the effect of amino group on adsorption. The adsorption capacity of p-nitrophenol on NA-01A increased noticeably by comparison with the mother adsorbent. A linear relationship was observed between the adsorption capacity of NA-01A caused by amination and the equilibrium concentration of p-nitrophenol in aqueous solution, which can be reasonably elucidated by a modified isotherm equation based on the Freundlich model. The breakthrough curves on both adsorbents were experimentally determined and also predicted by the mathematic model based on the non-linear wave propagation theory and the corresponding isotherm model. The model based on the Freundlich model gave a good prediction of the breakthrough curve on NA-01, but a deviation occurred for NA-01A when C/C0 less than 0.3. However, another model based on the modified isotherm equation provided a better prediction for the breakthrough curves on NA-01A. PMID- 16713090 TI - Secretory vesicles in live cells are not free-floating but tethered to filamentous structures: a study using photonic force microscopy. AB - It is well established that actin and microtubule cytoskeletal systems are involved in organelle transport and membrane trafficking in cells. This is also true for the transport of secretory vesicles in neuroendocrine cells and neurons. It was however unclear whether secretory vesicles remain free-floating, only to associate with such cytoskeletal systems when needing transport. This hypothesis was tested using live pancreatic acinar cells in physiological buffer solutions, using the photonic force microscope (PFM). When membrane-bound secretory vesicles (0.2-1.2 microm in diameter) in live pancreatic acinar cells were trapped at the laser focus of the PFM and pulled, they were all found tethered to filamentous structures. Mild exposure of cells to nocodazole and cytochalasin B, disrupts the tether. Immunoblot analysis of isolated secretory vesicles, further demonstrated the association of actin, myosin V, and kinesin. These studies demonstrate for the first time that secretory vesicles in live pancreatic acinar cells are tethered and not free-floating, suggesting that following vesicle biogenesis, they are placed on their own railroad track, ready to be transported to their final destination within the cell when required. This makes sense, since precision and regulation are the hallmarks of all cellular process, and therefore would hold true for the transport and localization of subcellular organelles such as secretory vesicles. PMID- 16713091 TI - Contributions to the contrast in experimental high-angle annular dark-field images. AB - This paper reports on a study of the contributions to the image contrast of high angle annular dark field (HAADF) images acquired in scanning transmission electron microscopy. Experimental HAADF images were obtained from a model system consisting of an epitaxial perovskite PbTiO3 film grown on a SrTiO3 single crystal. This sample allowed for the study of the intensities of a wide range of atomic numbers. The main objective of the paper was to quantify the influence of TEM foil thickness on the image contrast, but the effects of the annular detector inner angle and the probe forming lens focus were also studied. Sample thicknesses ranging from approximately 10 nm to more than 400 nm were investigated. The image contrast was relatively insensitive to changes in inner angle. The main impact of sample thickness was a rapid increase in a background intensity that contributed equally to the intensities of the atomic columns and the channels between them. The background intensity and its increase with thickness reflected the average atomic number of the crystal. Subtraction of the background intensity allowed for a quantitative interpretation of image contrast in terms of atomic numbers and comparison with multislice image simulations. The consequences for the analysis of interfaces in terms of atom column occupancies are discussed. PMID- 16713092 TI - Coherence measurements of zero-loss, plasmon-loss and phonon-loss electrons and their contribution to the Stobbs factor. PMID- 16713093 TI - Cell secretion machinery: studies using the AFM. AB - A new field in biology, 'nano-cell biology', has emerged from the successful use of force microscopy in understanding the structure and dynamics of cells and biomolecules, at nm resolution and in real time. Atomic force microscopy, in combination with conventional tools and approaches (electron microscopy, electrophysiology, X-ray diffraction, photon correlation spectroscopy, mass spectroscopy, biochemistry, and molecular biology), has revealed for the first time, the universal molecular machinery and mechanism of secretion in cells. Secretion occurs in all living cells and involves the delivery of intracellular products to the cell exterior. Secretory products are packaged and stored in membranous sacs or vesicles within the cell. When the cell needs to secrete these products, the secretory vesicles containing them, dock and fuse at plasma membrane-associated supramolecular structures called Porosome, to release their contents. Specialized cells for neurotransmission, enzyme secretion, or hormone release utilize a highly regulated secretory process. During secretion, swelling of secretory vesicles results in a build-up of intravesicular pressure, allowing expulsion of vesicular contents. The extent of vesicle swelling dictates the amount of vesicular contents expelled. The discovery of the porosome as the universal secretory machinery, its isolation, its structure and dynamics at nm resolution and in real time, its biochemical composition and functional reconstitution into artificial lipid membrane, have been determined. The molecular mechanism of secretory vesicle swelling, and the fusion of opposing bilayers, i.e., the fusion of secretory vesicle membrane at the base of the porosome membrane, has also been resolved. PMID- 16713094 TI - Mating success of two geographically distinct populations of Gulf Coast ticks, Amblyomma maculatum Koch. AB - Gulf Coast ticks collected from Refugio Co., TX and Osage Co., KS are reproductively compatible despite differences in genetic haplotypes, geographic separation and seasonal phenologies. Two heifers per mating combination (TX males x TX females, KS males x KS females, TX males x KS females, KS males x TX females) were each infested with 360 pairs of Gulf Coast ticks. Only mean pre oviposition and mean egg conversion efficiency index for the Texas male-Kansas female mating were significantly different (p<0.05) from other mating treatments. These females began oviposition 1-day later and used 4% less body mass toward egg production when compared to site-specific matings. However, the overall trend in reproductive performance of reciprocal tick matings was slightly lower than that of site-specific matings. There appear to be no pre-zygotic barriers to mating among Gulf Coast ticks from these Texas and Kansas populations. PMID- 16713095 TI - Comparison of scratching behaviour of growing pigs with sarcoptic mange before and after treatment, employing two distinct approaches. AB - In a closed pig breeding and finishing herd suffering from sarcoptic mange, two selected groups of pigs were filmed during a period of 10 days before and after treatment. The observation always commenced each hour and lasted for 15 min. Before treatment, observations was done round the clock, after treatment from 8:00 to 22:15. Before treatment the pens were stocked with 11 (pen A) and 10 (pen B) growing pigs (Large WhitexLandrace sows; 5 months old) with an average weight of approximately 70 kg examined for sarcoptic mange by skin scrapings and ELISA. The animals had never been treated with an acaricide or endectocide before. After 10 days, the pigs were treated twice (18 days interval) with Dectomax 1% solution for pigs (Pfizer, Austria) at a dose of 0.3 mg Doramectin i.m./kg body weight. After treatment, seven pigs were observed in both pens. Most scratching actions both before (83.1%) and after (94.5%) treatment were of one to 10 s. After treatment, the 10 s-scratching episodes decreased by 67.3% (from 21.2 to 6.9 mean SRE/pig), and the scratching actions of longer than 10 s by 91.7% (from 4.3 to 0.4 mean SRE/pig), such that the latter could be observed only occasionally after treatment. A distinct increase in scratching activity both before and after treatment could be observed primarily between 10:00 and 15:00. Significant differences of scratching and rubbing activity between before and after treatment could also be seen at midday. The interpretation of the scratching index values before and after the treatment were carried out according to Cargill et al. [Cargill, C., Davies, P., Carmichael, I., Hooke, F., Moore, M., 1994. Treatment of pigs with doramectin to control sarcoptic mange. Proceedings of the 13th IPVS Congress, Bangkok, Thailand, p. 238] with the maximum and minimal limiting values specified in the literature, and compared with calculations using the method described by Hollanders et al. [Hollanders, W., Harbers, A.H.M., Huige, J.C.M., Monster, P., Rambags, P.G.M., Hendrikx, W.M.L., 1995. Control of Sarcoptes scabiei var. suis with ivermectin: influence on scratching behaviour of fattening pigs and occurence of dermatitis at slaughter. Vet. Parasitol. 58, 117-127]. Depending on the methods used and the limiting values set, 6.7-34.6% of the observations before and 2.0-17.3% of the observations after treatment revealed a "strong evidence of mange" or a "suspicion of mange". All other observations indicated that the pigs were free from mange. PMID- 16713096 TI - Differential diagnosis of dog hookworms based on PCR-RFLP from the ITS region of their rDNA. AB - Species of Ancylostoma infecting dogs and sometimes humans are sympatric in many parts of the world. The establishment of a specific molecular diagnostic tool is important, not only to refine information for epidemiological studies, but also to evaluate the efficacy of vaccine programmes and assist in the development of specific drug treatments. The ITS region from 20 specimens of A. braziliense, collected from three separate geographical areas of Brazil, and from 10 specimens of A. caninum, collected from the same area in Brazil were sequenced and analyzed. Alignment of sequences showed that this gene is highly conserved. The intraspecific polymorphism for both species was less then 1%, whereas the interspecific polymorphism was 6.2, 7.3 and 9.4% between A. ceylanicum and A. braziliense; A. caninum and A. ceylanicum and A. ceylanicum and A. braziliense, respectively. Among the three species it was 12.3%. This revealed the ITS region as highly conserved and consequently a good molecular marker for diagnostic studies. In this work, four restriction enzymes were used in a PCR-RFLP using the ITS region of rDNA, to establish a differential diagnosis which discriminates between three Ancylostoma species, A. braziliense, A. caninum and A. ceylanicum. The best pattern was given by the HinfI enzyme, which produced different fragment sizes for each of the three species. Furthermore, the diagnostic tool differentiates DNA extracted directly from faeces of Ancylostoma-infected dogs. PMID- 16713097 TI - Interaction of Tritrichomonas foetus and the bovine oviduct in an organ culture model. AB - Tritrichomonas foetus is an extracellular parasite of the reproductive tract in cattle. The mechanism by which T. foetus causes abortion in cattle is largely unknown. There are no studies of infection in the cow oviducts, almost all published papers are related to vagina infection and few articles focusing on the uterus. The aim of the present study was to establish a working model of bovine oviduct epithelial cells and submit these cells to Tritrichomonas foetus interaction. Twenty bovine oviducts were obtained from cows at a commercial abattoir and T. foetus was injected through the isthmus into the oviduct lumen. The whole oviduct was analyzed by scanning and transmission electron microscopy. The results reported here demonstrate that: (1) fresh whole oviducts can be used as a good model to study parasite-host cell interaction; (2) cow oviduct epithelium has been shown to consist of two cell types: ciliated and nonciliated secretory cells, and T. foetus displayed great specificity for the nonciliated cells localized in the deeper oviduct folds; (3) T. foetus adheres as single separate cells, and maintains the flagella externalized; (4) differently from T. vaginalis, T. foetus does not change its shape during the adhesion process; and (5) oviduct cells exhibited morphological characteristics of apoptosis after trichomonadal interaction. PMID- 16713098 TI - Survey of helminths in adult sheep in Heilongjiang Province, People's Republic of China. AB - The prevalence of helminths in adult sheep was investigated in Heilongjiang Province, People's Republic of China between January 1999 and September 2003. A total of 326 adult sheep representing local breeds (Xingjiang Fine Wool Sheep, Dongbei Fine Wool Sheep) as well as introduced breeds (Merino and Charollais) from representative geographical locations in Heilongjiang Province were slaughtered and examined for the presence of helminths. The worms were examined, counted and identified to species according to existing keys and descriptions. A total of 26 helminth species were found representing 2 phyla, 3 classes, 13 families and 20 genera. All sheep were infected by more than one helminth species. Oesophagostomum columbianum, Haemonchus contortus and Trichostrongylus colubriformis were the most common nematode species, and Paramphistomum cervi, Orientobilharzia turkestanica and Fasciola hepatica were the most common trematode species, whereas the infection of adult sheep with cestodes was uncommon. The results of the present investigation provide relevant "base-line" data for Heilongjiang Province, China, for assessing the effectiveness of future control strategies against helminth infections in sheep. PMID- 16713099 TI - Detection of filarial parasites in domestic cats by PCR-RFLP of ITS1. AB - Lymphatic filariasis has been targeted by the World Health Organization (WHO) to be eliminated by the year 2020. In addition to chemotherapy and vector control, the control of reservoir hosts is necessary for the control program to succeed. Malayan filariasis, caused by Brugia malayi, is endemic in the South of Thailand where domestic cats serve as the major reservoir host. However, in nature, domestic cats also carry B. pahangi, Dirofilaria immitis and D. repens infections and it is difficult to distinguish the different filarial species from each other just by morphology. To assess the burden of filarial parasites, we performed a study on domestic cats in an endemic area of malayan filariasis in the Prasang district, of Surat Thani, a province in Southern Thailand. Together with Giemsa staining and acid phosphatase activity studies, we performed PCR-RFLP analysis on the first internal transcribed spacer (ITS1) region of ribosomal DNA (rDNA). PCR RFLP with Ase I could clearly differentiate between B. malayi, B. pahangi, D. immitis and D. repens. Out of the 52 cats studied, filarial parasites were identified in 5 (9.5%) cats, of which 4 (7.6%) were B. pahangi and 1 (1.9%) D. immitis. This PCR-RFLP technique detected two additional cats that were not detected by microscopy. The domestic cats are not an important host of B. malayi in this region. We could develop the PCR-RFLP assay test for differentiating filarial nematodes which can be applied to survey human, animal reservoir hosts and mosquito vectors in endemic areas. PMID- 16713100 TI - Peroxisome is a reservoir of intracellular calcium. AB - We have examined fura 2-loaded purified peroxisomes under confocal microscope to prove that this mammalian organelle is a store of intracellular calcium pool. Presence of calcium channel and vanadate sensitive Ca(2+)-ATPase in the purified peroxisomal membrane has been demonstrated. We have further observed that machineries to maintain calcium pool in this mammalian organelle are impaired during infection caused by Leishmania donovani. Results reveal that peroxisomes have a merit to play a significant role in the metabolism of intracellular calcium. PMID- 16713101 TI - Infections in burn patients--experience in a tertiary care hospital. AB - A retrospective study was conducted at the Kanchi Kamakoti Childs Trust Hospital to analyze the bacterial isolates from the wounds of patients admitted to the Burns Unit and to determine the sensitivity patterns of the commonly cultured organisms over a 6-year period, December 1998 to December 2004. A total of 535 samples were analysed. Single isolates were more common. Pseudomonas was the most commonly cultured organism followed by Staphylococcus aureus. Escherechia coli and Klebsiella were seen in equal numbers. ESBL rates were 37%. Antibiotic sensitivity patterns and periodic ESBL surveillance helped in selection of appropriate antibiotics. PMID- 16713102 TI - Cancer treatment reviews welcomes submission of the Cochrane Reviews. PMID- 16713103 TI - Current status of angiogenesis inhibitors combined with radiation therapy. AB - Angiogenesis inhibitors combined with cytotoxic chemotherapy have recently entered routine oncological practice. Several rationales exist for combining these agents with ionizing radiation, a primary curative cancer treatment, either in bimodal or trimodal fashion, i.e. with or without additional chemotherapy. More than 20 different anti-angiogenic agents have been studied in preclinical animal tumor models. This systematic review compares the results of preclinical studies published before February 2006. The combination of vascular endothelial growth factor (VEGF) inhibitors with irradiation consistently resulted in improved tumor growth delay (at least additive effects), despite different radiation schedules, drugs and doses, and combination regimens. Only two studies evaluated tumor control dose (TCD)50 as a measure of tumor cure (radiation dose yielding permanent local control in 50% of the tumors). While anti-VEGF receptor (VEGFR) antibody treatment improved the outcome, a VEGFR tyrosine kinase inhibitor showed negative results. For agents interfering with other pathways, the results are also not consistent, although most studies were positive. Trimodal approaches seem to improve tumor growth delay even further. Importantly, both radiotherapy schedule and sequence of the modalities in combined treatment may impact on the outcome. Hence, further preclinical studies examining these parameters need to be conducted. While preclinical research is ongoing, phase I and II clinical trials with bevacizumab, combretastatin A-4, thalidomide and different receptor tyrosine kinase inhibitors, usually combined with radio- and chemotherapy, have been designed. Early results suggest that acute toxicity is acceptable, planned surgery after such treatment is feasible, and that further evaluation of such combined modality treatment is warranted. PMID- 16713104 TI - Effects of nicotine on visuo-spatial selective attention as indexed by event related potentials. AB - Nicotine has been shown to specifically reduce reaction times to invalidly cued targets in spatial cueing paradigms. In two experiments, we used event-related potentials to test whether the facilitative effect of nicotine upon the detection of invalidly cued targets is due to a modulation of perceptual processing, as indexed by early attention-related event-related potential components. Furthermore, we assessed whether the effect of nicotine on such unattended stimuli depends upon the use of exogenous or endogenous cues. In both experiments, the electroencephalogram was recorded while non-smokers completed discrimination tasks in Posner-type paradigms after chewing a nicotine polacrilex gum (Nicorette 2 mg) in one session and a placebo gum in another session. Nicotine reduced reaction times to invalidly cued targets when cueing was endogenous. In contrast, no differential effect of nicotine on reaction times was observed when exogenous cues were used. Electrophysiologically, we found a similar attentional modulation of the P1 and N1 components under placebo and nicotine but a differential modulation of later event-related potential components at a frontocentral site. The lack of a drug-dependent modulation of P1 and N1 in the presence of a behavioral effect suggests that the effect of nicotine in endogenous visuo-spatial cueing tasks is not due to an alteration of perceptual processes. Rather, the differential modulation of frontocentral event related potentials suggests that nicotine acts at later stages of target processing. PMID- 16713105 TI - C fragment of tetanus toxin hybrid proteins evaluated for muscle-specific transsynaptic mapping of spinal motor circuitry in the newborn mouse. AB - We investigated whether the non-toxic C fragment of tetanus toxin (TTC) fused to either beta-galactosidase or green fluorescent protein could be utilized to transsynaptically trace muscle-specific spinal circuitry in the neonatal mouse after i.m. injection into a single hindlimb muscle. We found that even with careful low volume injection (0.2-1.0 microl) into a single muscle (medial gastrocnemius), the TTC hybrid proteins spread rapidly to many other hindlimb muscles and to trunk musculature such that retrograde labeling of motoneurons could not be constrained to a single motoneuron pool. Retrogradely labeled motoneurons in the lower lumbar segments harboring the medial gastrocnemius motoneuron pool were first observed two hours after the medial gastrocnemius injection. Within the next 10 h, additional lumbar and lower thoracic motoneurons became labeled, and punctate labeling in the neuropil surrounding the motoneurons appeared. Many of the TTC hybrid protein-labeled puncta in the neuropil co localized synaptotagmin, indicating that they represent presynaptic axon terminals onto motoneurons. Although this is consistent with retrograde transsynaptic passage, we found no evidence that the TTC hybrid proteins were transported further along premotor axons to label interneuron somata. The i.m. TTC injection procedure described here therefore provides an important tool for the study of presynaptic terminals onto motoneurons. However, additional technical modifications will be required to utilize TTC tracers for transsynaptic mapping of muscle-specific spinal motor circuitry in the neonatal mouse. We provide here a set of criteria for assessing the i.m. delivery of TTC tracers as a basis for future improvements in this technique. PMID- 16713107 TI - Sensory-motor interaction in primary hand cortical areas: a magnetoencephalography assessment. AB - Movement control requires continuous and reciprocal exchange of information between activities of motor areas involved in the task program execution and those elaborating proprioceptive sensory information. Our aim was to investigate the sensorimotor interactions in the region dedicated to hand control in healthy humans, focusing onto primary sensory and motor cortices, by selecting the time window at very early latencies. Through magnetoencephalographic recordings, we obtained a simultaneous assessment of sensory cortex activity modulation due to movement and of motor cortex activity modulation due to sensory stimulation, by eliciting a galvanic stimulation to the nerve (the median nerve) innervating a muscle (the opponens pollicis), at rest or during voluntary contraction. The primary sensory and motor cortices activities were investigated respectively through excitability in response to sensory stimulation and the cortico-muscular coherence. The task was performed bilaterally. A clear reduction of the cortico muscular coherence was found in the short time window following stimuli (between around 150-450 ms). In the same time period, the motor control of isometric contraction was preserved. This could suggest that cortical component of voluntary movement control was transiently mediated by neuronal firing rate tuning more than by cortico-muscular synchronization. In addition to the known primary sensory cortex inhibition due to movement, a more evident reduction was found for the component known to include a contribution from primary motor areas. Gating effects were lower in the dominant left hemisphere, suggesting that sensorimotor areas dominant for hand control benefit of narrowing down gating effects. PMID- 16713106 TI - Habituation to the test cage influences amphetamine-induced locomotion and Fos expression and increases FosB/DeltaFosB-like immunoreactivity in mice. AB - Pre-exposure to the testing cage (habituation or familiarization) is a common procedure aimed at reducing the interference of novelty-induced arousal and drug independent individual differences on neural and behavioral measures. However, recent results suggest that this procedure might exert a major influence on the effects of addictive drugs. The present experiments tested the effects of repeated exposure to a test cage (1 h daily for four consecutive days) on amphetamine-induced locomotion and Fos expression as well as on FosB/DeltaFosB like immunoreactivity in mice of the C57BL/6J and DBA/2J inbred strains that differ for the response to amphetamine, stress and novelty. Daily experiences with the test cage increased FosB/DeltaFosB-like immunoreactivity in the medial prefrontal cortex of both strains of mice and in the caudate of mice of the C57 strain, as reported for repeated stress in the rat. Moreover, previous habituation to the test cage reduced the locomotor response to a low dose of amphetamine only in DBA mice while it reduced amphetamine-induced Fos expression in medial-prefrontal cortex, dorsal caudate and the accumbens shell of mice of the C57 strain. These results demonstrate indexes of stress-like plasticity in the brains of mice exposed to a procedure of familiarization to the testing environment. Moreover, they suggest that the procedure of daily familiarization influences the pattern of brain Fos expression induced by amphetamine. Finally, they indicate complex interactions between experience with the testing environment, genotype and drug. PMID- 16713108 TI - A role for glutamate and glia in the fast network oscillations preceding spreading depression. AB - The mechanism of the propagation of spreading depression is unclear. Classical theories proposed a self-maintained cycle fed by elevated potassium and/or glutamate in the extracellular space. Earlier we found in vivo a characteristic oscillatory field activity that is synchronous in a strip of tissue ahead of the oncoming wave of neuron depolarization and that occurs before the extracellular potassium level begins to rise [Herreras O, Largo C, Ibarz JM, Somjen GG, Marrin del Rio R (1994) Role of neuronal synchronizing mechanisms in the propagation of spreading depression in the in vivo hippocampus. J Neurosci 14:7087-7098]. We investigated here the possible participation of glutamate and the role of glia in the prodromal field oscillations using extra and intracellular recordings and pharmacological manipulations in rat hippocampal slices. As earlier shown in vivo, field oscillations propagated ahead of the negative potential shift covering distances of up to 1 mm. The oscillatory prodromals were initially subthreshold but then each wave became crowned by a population spike. The frequency of the oscillatory prodromals was variable among slices (80-115 Hz), but constant in individual slices. The blockade of ionotropic glutamate receptors decreased the frequency of prodromal oscillations, retarded spreading depression propagation, and shortened the duration of depolarization. Blocking the glutamate membrane transport increased the oscillatory frequency. The selective metabolic poisoning of astrocytes led to gradual disorganization of prodromal oscillations whose frequency first increased and then decreased. Also, the amplitude of the population spikes within the burst diminished as individual cells fired fewer action potentials, although still phase-locked with population spikes. The effects of glial metabolic impairment were observed within the period when neuron electrical properties were still normal, and were blocked by glutamate receptor antagonists. These data suggest that glutamate released from glial cells and possibly also from neurons has a role in the generation of oscillations and neuron firing synchronization that precede the spreading depression-related depolarization, but additional mechanisms are required to fully explain the onset and propagation of spreading depression. PMID- 16713110 TI - Abnormal proprioceptive-motor integration contributes to hypometric postural responses of subjects with Parkinson's disease. AB - Subjects with Parkinson's disease exhibit abnormally short compensatory steps in response to external postural perturbations. We examined whether: (1) Parkinson's disease subjects exhibit short compensatory steps due to abnormal central proprioceptive-motor integration, (2) this proprioceptive-motor deficit can be overcome by visual-motor neural circuits using visual targets, (3) the proprioceptive-motor deficit relates to the severity of Parkinson's disease, and (4) the dysfunction of central dopaminergic circuits contributes to the Parkinson's disease subjects' proprioceptive-motor deficit. Ten Parkinson's disease subjects and 10 matched control subjects performed compensatory steps in response to backward surface translations in five conditions: with eyes closed, with eyes open, to a remembered visual target, to a target without seeing their legs, and to a target while seeing their legs. Parkinson's disease subjects were separated into a moderate group and a severe group based on scores from the Unified Parkinson's Disease Rating Scale and were tested off and on their dopamine medication. Parkinson's disease subjects exhibited shorter compensatory steps than did the control subjects, but all subjects increased their step length when stepping to targets. Compared with the other subject groups, the severe Parkinson's disease subjects made larger accuracy errors when stepping to targets, and the severe Parkinson's disease subjects' step accuracy worsened the most when they were unable to see their legs. Thus, Parkinson's disease subjects exhibited short compensatory steps due to abnormal proprioceptive-motor integration and used visual input to take longer compensatory steps when a target was provided. In severe Parkinson's disease subjects, however, visual input does not fully compensate because, even with a target and unobstructed vision, they still exhibited poor step accuracy. Medication did not consistently improve the length and accuracy of the Parkinson's disease subjects' compensatory steps, suggesting that degeneration of dopamine circuits within the basal ganglia is not responsible for the proprioceptive-motor deficit that degrades compensatory steps in Parkinson's disease subjects. PMID- 16713109 TI - Role of p38 and inducible nitric oxide synthase in the in vivo dopaminergic cells' degeneration induced by inflammatory processes after lipopolysaccharide injection. AB - Accumulating evidences suggest that neuroinflammation is involved in the progressive death of dopaminergic neurons in Parkinson's disease. Several studies have shown that intranigral injection of lipopolysaccharide induces inflammation in the substantia nigra leading to death of tyrosine hydroxylase-positive cells. To better understand how the inflammatory response gives rise to neurotoxicity we induced inflammation in substantia nigra by injecting lipopolysaccharide. The damage of substantia nigra dopaminergic neurons was evaluated by immunohistochemistry, reverse transcription-PCR and Western blot analysis of tyrosine hydroxylase. In parallel, activation of microglial cells, a hallmark of inflammation in CNS, was revealed by immunohistochemistry. Similarly the expression of molecules involved in the inflammatory response and apoptotic pathway was also tested, such as cytokines (tumor necrosis factor-alpha, interleukin-1beta, interleukin-6), inducible nitric oxide synthase and caspase 11. Tyrosine hydroxylase expression (both mRNA and protein) started to decrease around 3 days post-injection. At the mRNA level, our results showed that the cytokines expression peaked shortly (3-6 h) after lipopolysaccharide injection, followed by the induction of inducible nitric oxide synthase and caspase-11 (14 h). However, inducible nitric oxide synthase protein peaked at 24 h and lasted for 14 days. The lipopolysaccharide-induced loss of substantia nigra dopaminergic neurons was partially inhibited by co-injection of lipopolysaccharide with S methylisothiourea, an inducible nitric oxide synthase inhibitor. Co-injections of lipopolysaccharide with SB203580, a p38 MAP kinase inhibitor, reduced inducible nitric oxide synthase and caspase-11 mRNA expression, and also rescued dopaminergic neurons in substantia nigra. In summary, this is the first report to describe in vivo the temporal profile of the expression of these inflammatory mediators and proteins involved in dopaminergic neuronal death after intranigral injection of lipopolysaccharide. Moreover data strongly support that lipopolysaccharide-induced dopaminergic cellular death in substantia nigra could be mediated, at least in part, by the p38 signal pathway leading to activation of inducible nitric oxide synthase and caspase-11. PMID- 16713111 TI - The role of the rat prelimbic/infralimbic cortex in working memory: not involved in the short-term maintenance but in monitoring and processing functions. AB - Contrary to human and primate, working memory in the rodent is usually considered as a simple short term memory buffer and mainly investigated using delayed response paradigms. The aim of the present study was to further investigate the role of the rat prelimbic/infralimbic cortex in different spatial delayed tasks in order to dissociate its involvement in temporary storage from other information processes, such as behavioral flexibility and attention. In experiment 1 rats were trained in a standard elimination win-shift task in a radial-arm maze after which a 1-min delay was inserted mid trial. Prelimbic/infralimbic lesions induced only a transient disruption of performance following introduction of the delay. In experiment 2, rats were trained directly in a win-shift task with a 5-min delay that was subsequently extended to 30 min. Prelimbic/infralimbic lesions did not significantly affect behavior. Nevertheless, transient disruptions of performance (correlated with lesion extent) were noted repeatedly in lesioned rats when sets of interfering events were presented. The present findings indicate that prelimbic/infralimbic cortex is not directly involved in the short term maintenance of specific information but is implicated when changes, such as sudden introduction of a delay or exposure to unexpected interfering events, alter the initial situation. It appears that working memory in rodents should be considered, as in humans and primates, to encompass both storage and monitoring functions. The present results along with previous ones strongly suggest that prelimbic/infralimbic cortex is not involved in the temporary on-line storage but rather in the control of information required to prospectively organize the ongoing action. PMID- 16713112 TI - Intrathecal sensory neuron-specific receptor agonists bovine adrenal medulla 8-22 and (Tyr6)-gamma2-MSH-6-12 inhibit formalin-evoked nociception and neuronal Fos like immunoreactivity in the spinal cord of the rat. AB - The finding that sensory neuron-specific G-protein-coupled receptor mRNA is solely expressed in small primary sensory neurons suggests involvement of the receptor in nociceptive modulation. The present study was designed to assess effects of intrathecal administration of bovine adrenal medulla 8-22 and (Tyr6) gamma2-MSH-6-12, selective sensory neuron-specific receptor agonists, on nocifensive behaviors and expression of spinal c-Fos-like immunoreactivity evoked by intraplantar injection of 2.5% formalin in rats. The agonists were administered 10 min before (pretreatment) and/or after (post-treatment) injection of formalin. Pretreatment with bovine adrenal medulla 8-22 dose-dependently (3, 10 and 30 nmol) decreased time lifting and licking the paw mainly in the second phase. Intrathecal bovine adrenal medulla 8-22 (30 nmol) remarkably suppressed nocifensive behaviors in the first and second phases and the expression of formalin-evoked c-Fos-like immunoreactivity in laminae I-II and V-VI of the spinal dorsal horn at L4-5. Moreover, naloxone (20 microg, intrathecal) failed to antagonize the inhibitory effects of bovine adrenal medulla 8-22. Post-treatment with bovine adrenal medulla 8-22 also exerted inhibition on the second phase behaviors in a dose-dependent manner with a similar efficacy observed in pretreatment groups. Furthermore, post-treatment with (Tyr6)-gamma2-MSH-6-12 (0.5, 1.5 and 5 nmol) also suppressed formalin-evoked nocifensive behaviors in the second phase and c-Fos-like immunoreactivity in the spinal dorsal horn similar with bovine adrenal medulla 8-22. Our results suggest that sensory neuron specific receptor may play an important role in modulation of spinal nociceptive transmission. This is the first to demonstrate that activation of sensory neuron specific receptor produces analgesia in the persistent pain model. PMID- 16713114 TI - Glucose and lactate are equally effective in energizing activity-dependent synaptic vesicle turnover in purified cortical neurons. AB - This study examines the role of glucose and lactate as energy substrates to sustain synaptic vesicle cycling. Synaptic vesicle turnover was assessed in a quantitative manner by fluorescence microscopy in primary cultures of mouse cortical neurons. An electrode-equipped perfusion chamber was used to stimulate cells both by electrical field and potassium depolarization during image acquisition. An image analysis procedure was elaborated to select in an unbiased manner synaptic boutons loaded with the fluorescent dye N-(3 triethylammoniumpropyl)-4-(4-(dibutylamino)styryl)pyridinium dibromide (FM1-43). Whereas a minority of the sites fully released their dye content following electrical stimulation, others needed subsequent K(+) depolarization to achieve full release. This functional heterogeneity was not significantly altered by the nature of metabolic substrates. Repetitive stimulation sequences of FM1-43 uptake and release were then performed in the absence of any metabolic substrate and showed that the number of active sites dramatically decreased after the first cycle of loading/unloading. The presence of 1 mM glucose or lactate was sufficient to sustain synaptic vesicle cycling under these conditions. Moreover, both substrates were equivalent for recovery of function after a phase of decreased metabolic substrate availability. Thus, lactate appears to be equivalent to glucose for sustaining synaptic vesicle turnover in cultured cortical neurons during activity. PMID- 16713113 TI - Role of interleukin-6 in lipopolysaccharide-induced brain injury and behavioral dysfunction in neonatal rats. AB - There are increasing data in support of the hypothesis that inflammatory cytokines are involved in neonatal white matter damage. Despite extensive study of the proinflammatory cytokines tumor necrosis factor-alpha and interleukin 1beta, the role of interleukin-6 in the development of white matter damage is largely unknown. In the present study, the role(s) of interleukin-6 in mediating lipopolysaccharide-induced brain injury and behavioral changes was investigated by the intracerebral injection of lipopolysaccharide with interleukin-6 neutralizing antibody in the 5-day-old rat brain. Brain injury was examined in brain sections at postnatal day 8 and postnatal day 21. Behavioral tests including righting reflex, wire hanging maneuver, cliff avoidance, locomotor activity, gait analysis, responses in the elevated plus-maze and passive avoidance were performed from postnatal day 3 to postnatal day 21. Changes in astroglia, microglia and oligodendrocytes were studied using immunohistochemistry in the postnatal day 21 rat brain. Our results show that interleukin-6 antibody attenuated lipopolysaccharide-induced brain lateral ventricle dilation and improved neurobehavioral performance. Interleukin-6 antibody also suppressed lipopolysaccharide-induced astrogliosis and microglial activation, and increased the number of oligodendrocytes in white matter. However, no changes of tumor necrosis factor-alpha and interleukin-1beta were detected. In contrast, no histopathological changes and glial activation were observed in rats injected with only interleukin-6. The present study indicates that the contribution to brain injury by interleukin-6 depends on its interaction with other lipopolysaccharide-induced agents and not on interleukin-6 alone. PMID- 16713115 TI - Cardiorespiratory fitness and acute aerobic exercise effects on neuroelectric and behavioral measures of action monitoring. AB - Cardiorespiratory fitness and acute aerobic exercise effects on cognitive function were assessed for 28 higher- and lower-fit adults during a flanker task by comparing behavioral and neuroelectric indices of action monitoring. The error related negativity, error positivity, and N2 components, as well as behavioral measures of response speed, accuracy, and post-error slowing were measured following a 30-minute acute bout of treadmill exercise or following 30-minutes of rest. A graded maximal exercise test was used to measure cardiorespiratory fitness by assessing maximal oxygen uptake. Results indicated that higher-fit adults exhibited reduced error-related negativity amplitude, increased error positivity amplitude, and increased post-error response slowing compared with lower-fit adults. However, acute exercise was not related to any of the dependent measures. These findings suggest that cardiorespiratory fitness, but not acute aerobic exercise, may be beneficial to behavioral and neuroelectric indices of action monitoring following errors of commission by increasing top-down attentional control. PMID- 16713116 TI - GABAergic mechanisms in regulating the activity state of substantia nigra pars reticulata neurons. AB - Substantia nigra reticulata is the major output structure of the basal ganglia involved in somatosensory integration and organization of movement. While previous work in vitro and in anesthetized animal preparations suggests that these neurons are autoactive and points to GABA as a primary input regulating their activity, single-unit recording coupled with iontophoresis was used in awake, unrestrained rats to further clarify the role of tonic and phasic GABA input in maintenance and fluctuations of substantia nigra reticulata neuronal activity under physiologically relevant conditions. In contrast to glutamate, which was virtually ineffective at stimulating substantia nigra reticulata neurons in awake rats, all substantia nigra reticulata neurons tested were inhibited by iontophoretic GABA and strongly excited by bicuculline, a GABA-A receptor blocker. The GABA-induced inhibition had short onset and offset latencies, a fading response pattern (a rapid decrease in rate followed by its relative restoration), and was independent of basal discharge rate. The bicuculline-induced excitation was inversely related to discharge rate and current (dose)-dependent in individual units. However, the average discharge rate during bicuculline applications at different currents increased to a similar plateau ( approximately 60 impulses/s), which was about twice the mean basal rates. The excitatory effects of bicuculline were phasically inhibited or completely blocked by brief GABA applications and generally mimicked by gabazine, another selective GABA antagonist. These data as well as neuronal inhibitions induced by nipecotic acid, a selective GABA uptake inhibitor, suggest that substantia nigra reticulata neurons in awake, quietly resting conditions are under tonic, GABA-mediated inhibition. Therefore, because of inherent autoactivity and specifics of afferent inputs, substantia nigra reticulata neurons are very sensitive to phasic alterations in GABA input, which appears to be the primary factor determining fluctuations in their activity states under physiological conditions. While these cells are relatively insensitive to direct activation by glutamate, and resistant to a continuous increase in GABA input, they appear to be very sensitive to a diminished GABA input, which may release them from tonic inhibition and determine their functional hyperactivity. PMID- 16713117 TI - Treatment-induced prevention of learning deficits in newborn mice with brain lesions. AB - Perinatal brain injuries often result in irreversible learning disabilities, which manifest in early childhood. The molecular and cellular mechanisms of these injuries and potential pharmacological treatments are emerging, chiefly from studies in newborn rodents. In newborn mice, experimentally induced lesions can be dramatically reduced by appropriate neuroprotective treatments. However, the early effectiveness of these treatments in preserving cognition remained unknown. Here, we addressed this issue by using intracerebral ibotenate to induce excitotoxic brain lesions in 5-day-old mice (postnatal day 5). On postnatal days 6-7, we tested spontaneous preference for maternal odors, as an index of odor memory, and conditioned preference for an artificial odor previously paired with stroking, as an index of associative learning. Brain-lesioned newborn mice showed normal general status and preference for maternal odors. In contrast, odor conditioning was severely impaired. A previous study showed that fructose 1,6 biphosphate acted as a neuroprotective agent which significantly reduced neocortical lesion size. In the present study, treating the newborn mice with fructose 1,6-biphosphate 15 min before the ibotenate injection reduced neocortical lesion size and restored conditioning. This demonstrates, for the first time, that neuroprotective treatment can protect some features of early cognition. PMID- 16713118 TI - Innervation and functional changes in mesenteric perivascular calcitonin gene related peptide- and neuropeptide Y-containing nerves following topical phenol treatment. AB - We have previously shown that age-related reduction of innervation and function in mesenteric perivascular calcitonin gene-related peptide-containing vasodilator nerves takes place in spontaneously hypertensive rats. The present study was performed to investigate innervation and functional changes in perivascular calcitonin gene-related peptide- and adrenergic neuropeptide Y-containing nerves after topical treatment with phenol, which damages nerve fibers, around the rat superior mesenteric artery. Under pentobarbital-Na anesthesia, 8-week-old Wistar rats underwent in vivo topical application of phenol (10% phenol in 90% ethanol) or saline (sham rats) to the superior mesenteric artery proximal to the bifurcation of the abdominal aorta. After the treatment, the animals were subjected to immunohistochemistry of the 3rd branch of small arteries proximal to the intestine and to vascular responsiveness testing on day 3 through day 14. The innervation levels of calcitonin gene-related peptide-like immunoreactivity containing fibers and neuropeptide Y-like immunoreactivity containing fibers were markedly reduced on day 3 to day 14 and on day 5 to day 14 after the treatment, compared with those in sham-operated rats, respectively. In perfused mesenteric vascular beds isolated from phenol-treated rats, adrenergic nerve-mediated vasoconstriction and calcitonin gene-related peptide nerve-mediated vasodilation in response to periarterial nerve stimulation (2-12 Hz) were significantly decreased on day 3 and day 7. Neurogenic release of norepinephrine in phenol treated rats on day 7 was significantly smaller that that in sham-operated rats. Nerve growth factor content in the mesenteric arteries of phenol-treated rats was significantly lower than that in sham-operated rats. Administration of nerve growth factor using osmotic mini-pumps for 7 days after the phenol treatment resulted in greater density of calcitonin gene-related peptide- and neuropeptide Y-like immunoreactivity fibers than in phenol-treated rats and restored decreased vascular responses to periarterial nerve stimulation. These results suggest that topical phenol-treatment of the mesenteric artery effectively induces functional denervation of perivascular nerves, which can be prevented or reversed by nerve growth factor treatment. PMID- 16713119 TI - Corticotropin-releasing factor in the dorsal raphe elicits temporally distinct serotonergic responses in the limbic system in relation to fear behavior. AB - The neurotransmitters serotonin and corticotrophin-releasing factor are thought to play an important role in fear and anxiety behaviors. This study aimed to determine the relationship between corticotrophin-releasing factor-evoked changes in serotonin levels within discrete regions of the limbic system and the expression of fear behavior in rats. The effects of corticotrophin-releasing factor administration to the serotonin cell body regions of the dorsal raphe nucleus on fear behavior, behavioral activity, and extracellular serotonin levels were assessed in freely moving rats with microdialysis probes implanted into the central nucleus of the amygdala and the medial prefrontal cortex. Infusion of corticotrophin-releasing factor (0.5 microg) into the dorsal raphe rapidly induced freezing behavior, which was positively correlated with an immediate increase in serotonin release in the central nucleus of the amygdala. In contrast, cessation of freezing behavior correlated with a delayed and prolonged increase in serotonin release within the medial prefrontal cortex. Our findings suggest that corticotrophin-releasing factor-induced freezing behavior is associated with regionally and temporally distinct serotonergic responses in the limbic system that may reflect differing roles for these regions in the expression of fear/anxiety behavior. PMID- 16713120 TI - Immunocytochemical colocalization of GABA-B receptor subunits in gonadotropin releasing hormone neurons of the sheep. AB - GABA has been shown to play an important role in the control of gonadotropin releasing hormone (GnRH) and luteinizing hormone secretion in many mammals. In sheep, seasonal differences in the ability of GABA-B receptor antagonists to alter pulsatile luteinizing hormone secretion have led to the hypothesis that this receptor subtype mediates the increased inhibitory effects of estradiol on GnRH and luteinizing hormone pulse frequency seen during the non-breeding season (anestrus). The aim of the present study was to use multiple-label immunocytochemistry to determine if ovine GnRH neurons contain the GABA-B receptor subunits R1 and/or R2, and to determine whether there are seasonal differences in the colocalization of these subunits in GnRH neurons. A majority of GnRH cells in the preoptic area, anterior hypothalamic area, and medial basal hypothalamus of both breeding season and anestrous ewes contained either GABA-B R1 or R2 subunits; a subset of GnRH neurons in breeding season (42%) and anestrous ewes (60%) contained both subunits. In contrast to colocalization within cell bodies, GnRH fibers in the median eminence did not colocalize GABA-B receptor subunits. Although the percentage of GnRH neurons expressing GABA-B receptor subunits tended to be higher in anestrus than in the breeding season, there were no significant seasonal differences in R1 and R2 subunit colocalization in GnRH cell bodies. Thus, while GABA may act directly on GnRH cell bodies via GABA-B receptors in the sheep, any role that GABA-B receptors may play in seasonal reproductive changes is likely mediated by other neurons afferent to GnRH cells. PMID- 16713121 TI - Long-term fate and distribution of newborn cells in the adult mouse olfactory bulb: Influences of olfactory deprivation. AB - The adult subventricular zone produces neuroblasts that migrate to the main olfactory bulb, where they differentiate into interneurons in the glomerular and granular layers. Using bromodeoxyuridine labeling, the survival of newborn cells was assessed in these two layers of the MOB in control mice and in mice unilaterally deprived from sensory input by naris occlusion. In control main olfactory bulbs, bromodeoxyuridine-positive cell density decreased about 70% between 15 and 180 days post-bromodeoxyuridine administration but earlier in the glomerular layer than in the granular layer. At all time points examined, newborn cell density was higher in the deep granular layer than in the superficial granular layer. Occlusion started at the age of 2 months and lasted for 15, 30, 45, 60 or 180 days. The newborn cell survival was similarly reduced in both layers by occlusion, during a critical period 15 and 45 days post-occlusion. Interestingly, olfactory deprivation decreased bromodeoxyuridine-positive cell density in the deep granular layer only, indicating a greater dependence of cell fate on sensory input in this sub-layer. Neuronal differentiation was assessed in the granular layer and glomerular layer by multiple double-labeling 45 days post bromodeoxyuridine-injections, the time point at which the proportion of bromodeoxyuridine-positive cells expressing a neuronal marker reached approximately 85% in the granular layer and approximately 50% in the glomerular layer. Naris occlusion did not significantly affect these proportions. Taken together, our results reveal that the survival of newborn cells has a different time course in the glomerular layer and in the granular layer, but is similarly decreased in each layer by olfactory deprivation. In addition, our data suggest a functional heterogeneity of neurogenesis within the granular layer. PMID- 16713122 TI - Duration of cannabis use--a novel phenotype? AB - Although cannabis is the most commonly used illicit drug, duration of cannabis use is typically short, with many of those who initiate cannabis use ceasing use by their late twenties. In this paper we analyze data from a volunteer Australian cohort of 6265 male and female twins to examine whether the duration of cannabis use is an informative phenotype for future genetic analyses. Genetic modeling indicated: (a) moderate genetic influences on duration of cannabis use in both males (41%; 95% CI=31-51) and females (55%; 95% CI=46-63); (b) strong genetic influences on cannabis dependence in both males (72%, 95% CI=61-81) and females (62%, 95% CI=48-74); (c) no evidence of shared environmental influences on duration of cannabis use or on cannabis dependence in either males or females. Importantly, this model fitting indicated that a substantial component of genetic influences (rg=.90, 95% CI=.77-.99 (males); .70, 95% CI=.57-.83 (females)) on duration of cannabis use was shared with those influencing liability to cannabis dependence. While there were high genetic correlations in both women and men, lifetime duration of cannabis may be uniquely informative in assessing components of liability to cannabis use. PMID- 16713123 TI - Psychiatric comorbidity and progression in drug use in adult male twins: implications for the design of genetic association studies. AB - Psychiatric comorbidity with drug dependence has been widely documented. In the present study, we reanalyze DSM-III-R diagnostic data on middle-aged male twin pairs from the VETR study using latent class methods. We identify four subtypes based on 15 diagnostic categories. We then show that these subtypes are strongly associated with differential rates of transitions in drug use histories, with increased risks in relatives for depression, alcohol, drug and ASPD, as well as with a variety of non-normative and deviant behaviors in youth and in adulthood. We use the result of these analyses to show how the use of a particular drug disorder phenotype for selecting cases could impact final sample composition. PMID- 16713126 TI - Moderators of delay tolerance in treatment-seeking cocaine users. AB - A substantial amount of attrition in cocaine dependence treatment studies occurs between the initial telephone contact and the first evaluative clinic visit. While decreasing the wait to first visit can significantly reduce pre-intake attrition (PIA), little is known about other factors that moderate delay tolerance for first clinic visit. The current report uses data from 833 subjects who completed a first-contact telephone interview prior to an intake evaluation visit for cocaine use treatment research. Hierarchical logistic regression was used to assess three successive models to predict PIA, with the most inclusive model testing interactions between delay interval and seven predictors: age, gender, treatment motivation, recency of cocaine, alcohol, and tobacco use, and self-reported depression. Consistent with previous reports, greater delay to first clinic visit predicted PIA. However, no evidence for the moderating role of the selected factors was found. Overall, the utility of the logistic models, built on basic demographic and psychiatric factors, was poor, as evaluated using receiver-operator characteristic curves. Alternative factors must be examined to identify predictors that will increase probability of initial enrolment in cocaine-dependence clinical trials. PMID- 16713125 TI - Does expectancy affect alcohol absorption? AB - Many factors influence alcohol absorption, yet few studies have addressed the issue of whether or not experimental manipulations themselves may affect alcohol absorption. The current balanced placebo design study comparing the expectancy effects of root beer and non-alcoholic beer vehicles resulted in significantly lower blood alcohol levels in the root beer condition than in the beer condition even though alcohol doses were the same. Two possible explanations are discussed; differences in expectancy may have affected absorption, or fructose in the root beer may have slowed absorption of alcohol relative to the maltose in beer. The literature does not provide strong evidence for either of the hypotheses. The implication of this study's results is that alcohol absorption rate may be an important source of confounding effects in behavioral research in the laboratory, because it may be affected by beverages or other experimental conditions. PMID- 16713127 TI - Combination of sulfated oligosaccharides and chemotherapy: a novel therapeutic strategy for cancer. PMID- 16713128 TI - Intestinal adenocarcinoma arising in a mature cystic teratoma of the ovary: a case report. AB - Gastrointestinal adenocarcinoma arising in mature cystic teratomas of the ovary is extremely rare. We report a case of well-differentiated intestinal adenocarcinoma arising in a mature cystic teratoma of the ovary in a 77-year-old woman, presenting as acute abdomen with ovarian torsion. An immunohistochemical study revealed expression of CK20 and CK7, and the tumor was also positive for MUC2. The patient had no evidence of disease after 12 months of follow-up. PMID- 16713124 TI - Is inhalant use a risk factor for heroin and injection drug use among adolescents in the United States? AB - PURPOSE: We examined whether inhalant use was associated with heroin and injection drug use (IDU) among adolescents aged 12 to 17 in the United States. METHODS: Data were drawn from the 2002/2003 administrations of the National Survey on Drug Use and Health (NSDUH). We conducted logistic regression analyses to estimate associations of inhalant use with heroin use, heroin injection, and IDU, respectively, among adolescent drug users (N=8161). RESULTS: Approximately 30.9% of adolescents had ever used at least one illicit drug. More than one-fifth (22.2%) of adolescents were past-year or recent drug users. Among past-year adolescent drug users, 1.4% had progressed to heroin use and 1.2% reported IDU. Adolescents who had used inhalants and marijuana were 2.8 and 2.9 times as likely as adolescents who had used marijuana but not inhalants to report heroin use and any IDU, respectively. Adolescents who had used inhalants or other drugs but not marijuana were unlikely to use heroin. However, inhalant users, irrespective of their marijuana use histories, had greater odds of IDU than drug users who had not used inhalants. Adolescent drug users who were females, school dropouts, whites, or delinquents had significantly increased odds of heroin use and IDU. Cigarette smoking before the age of 15 was strongly associated with heroin use, and a history of foster care placement was associated with IDU. CONCLUSIONS: This national study of American adolescents identifies several subgroups of recent drug users, such as females, school dropouts, and youth who have used inhalants and marijuana, which have substantially increased odds of heroin use and IDU. Screening, prevention, and treatment interventions targeted to these groups might reduce medical and social complications of heroin use and IDU. PMID- 16713129 TI - Evaluation of the wound healing properties of Acalypha langiana in diabetic rats. AB - Acalypha langinia is a well-known plant in the traditional medicine. Based on its traditional use, this plant was selected for evaluation of its wound healing potential. Topical application twice a day for 7 days of 0.05%, 0.1%, 0.2%, 0.4% and 0.5% sterile solution of aqueous extract from leaves of A. langinia significantly increased the healing process. PMID- 16713130 TI - Antimicrobial activity of the Derris elliptica, Derris indica and Derris trifoliata extractives. AB - Various parts of Derris elliptica, Derris indica and Derris trifoliata on fractionation with a number of solvents (petrol, dichloromethane, ethyl acetate, butanol and methanol) gave fractions which demonstrated a varied level of broad spectrum antibacterial activity. Good activity was exhibited by the methanol fractions of the leaves and root heart-wood, petrol, butanol and methanol fractions of the root bark of D. indica and petrol and ethyl acetate fractions of D. trifoliata. None of the plants showed antifungal activity. PMID- 16713131 TI - Hypoglycemic effect of Bauhinia cheilandra in rats. AB - The extract of the methanolic leaves of Bauhinia cheilandra (BC) was tested on glucose loaded and alloxan-induced diabetic rats. In both tests, the methanolic extract at doses of 300, 600, and 900 mg/kg, has shown a statistically significant and considerable hypoglycemic activity. PMID- 16713132 TI - Induction of two cytochrome P450 genes, Cyp6a2 and Cyp6a8, of Drosophila melanogaster by caffeine in adult flies and in cell culture. AB - To examine whether caffeine, the most widely used xenobiotic compound, would induce insect cytochrome P450 or CYP gene expression, upstream DNA fragments of Cyp6a2 (0.12, 0.26, 0.52 and 0.98-kb) and Cyp6a8 (0.06, 0.1, 0.2, 0.5 and 0.8-kb) genes of Drosophila melanogaster were individually fused to the firefly luciferase (luc) reporter gene. Promoter activities of these constructs were examined in Drosophila SL-2 cells using luciferase assays. Activity of 0.2- and 0.8-kb upstream DNA of Cyp6a8 was also measured in transgenic female flies. When these flies were treated with 2 mM pure caffeine or Vivarin caffeine, both DNA fragments showed a 4-5-fold induction of promoter activity. Endogenous Cyp6a8 and Cyp6a2 genes in these flies also showed caffeine-induced expression. In addition, both 0.2- and 0.8-kb DNAs showed differential basal and caffeine-induced activity in head, ovaries, gut, cuticle plus fat body and malpighian tubules. However, in all tissues 0.8-kb DNA always showed higher basal and caffeine-induced activities compared to the 0.2-kb DNA, suggesting that the additional DNA present in the 0.8 kb fragment has sequences that enhance both activities. In SL-2 cells, all reporter constructs of each Cyp6 gene showed significantly higher basal activity than the empty vector. Sequences that boost basal activity are located in -265/ 129 and -983/-522 DNA of Cyp6a2, and -199/-109 and -491/-199 DNA of Cyp6a8 genes. While the 0.12- and 0.1-kb upstream DNAs of Cyp6a2 and Cyp6a8 genes respectively did not show caffeine-inducibility in SL-2 cells, the longest upstream DNA of each gene gave the highest level of induction. Caffeine-responsive sequences are not clustered at one place; they appear to be dispersed in -983/-126 and -761/ 109 regions of Cyp6a2 and Cyp6a8 genes which also contain many binding sites for activator protein 1 (AP1) and cyclic AMP response element binding protein (CRE BP). Significance of these binding sites in caffeine-inducibility has been discussed. PMID- 16713134 TI - Venom gland EST analysis of the saw-scaled viper, Echis ocellatus, reveals novel alpha9beta1 integrin-binding motifs in venom metalloproteinases and a new group of putative toxins, renin-like aspartic proteases. AB - Echis ocellatus is the most medically important snake in West Africa. However, the composition of its venom and the differential contribution of these venom components to the severe haemorrhagic and coagulopathic pathology of envenoming are poorly understood. To address this situation we assembled a toxin transcriptome based upon 1000 expressed sequence tags (EST) from a cDNA library constructed from pooled venom glands of 10 individual E. ocellatus. We used a variety of bioinformatic tools to construct a fully annotated venom-toxin transcriptome that was interrogated with a combination of BLAST annotation, gene ontology cataloguing and disintegrin-motif searching. The results of these analyses revealed an unusually abundant and diverse expression of snake venom metalloproteinases (SVMP) and a broad toxin-expression profile including several distinct isoforms of bradykinin-potentiating peptides, phospholipase A(2), C-type lectins, serine proteinases and l-amino oxidases. Most significantly, we identified for the first time a conserved alpha(9)beta(1) integrin-binding motif in several SVMPs, and a new group of putative venom toxins, renin-like aspartic proteases. PMID- 16713133 TI - Unraveling the 'DEAD-box' helicases of Plasmodium falciparum. AB - The causative agent for the most fatal form of malaria, Plasmodium falciparum, has developed insecticide and drug resistance with time. Therefore combating this disease is becoming increasingly difficult and this calls for finding alternate ways to control malaria. One of the feasible ways could be to find out inhibitors/drugs specific for the indispensable enzymes of malaria parasite such as helicases. These helicases, which contain intrinsic nucleic acid-dependent ATPase activity, are capable of enzymatically unwinding energetically stable duplex nucleic acids into single-stranded templates and are required for all the nucleic acid transactions. Most of the helicases contain a set of nine extremely conserved amino acid sequences, which are called 'helicase motifs'. Due to the presence of the DEAD (Asp-Glu-Ala-Asp) in one of the conserved motifs, this family is also known as the 'DEAD-box' family. In this review, using bioinformatic approach, we describe the 'DEAD-box' helicases of malaria parasite P. falciparum. An in depth analysis shows that the parasite contains 22 full length genes, some of which are homologues of well-characterized helicases of this family from other organisms. Recently we have cloned and characterized the first member of this family, which is a homologue of p68 and is expressed during the schizont stage of the development of the parasite [Pradhan, A., Chauhan, V.S., Tuteja, R., 2005a. A novel 'DEAD-box' DNA helicase from Plasmodium falciparum is homologous to p68. Mol. Biochem. Parasitol. 140, 55-60.; Pradhan A., Chauhan V.S., Tuteja R., 2005b. Plasmodium falciparum DNA helicase 60 is a schizont stage specific, bipolar and dual helicase stimulated by PKC phosphorylation. Mol. Biochem. Parasitol. 144, 133-141.]. It will be really interesting to clone and characterize other members of the 'DEAD-box' family and understand their role in the replication and transmission of the parasite. These detailed studies may help to identify a parasite-specific enzyme, which could be a potential drug target to treat malaria. The various steps at which this probable drug can act are also discussed. PMID- 16713135 TI - Epidemiological approaches to safety investigations. AB - This paper considers the different approaches to post-authorisation safety monitoring of veterinary medicinal products that is essential to ensure confidence in their safety. Most safety testing is undertaken prior to granting of a marketing authorisation and is generally on a small scale. Field trials are usually much larger, but still involve relatively low numbers of animals compared to the number to which authorised products are administered. Safety testing is generally aimed at detecting common events; the numbers of animals used in the tests are too small for detection of all but the most common reactions. The efficiency of the tests depends on the frequency and severity of the adverse reaction and the ability to associate the adverse event with the product. The latter is affected by the period of time between administration and the event, as well as by its underlying frequency. Adverse reaction surveillance is critical in monitoring the safety of a marketed product. Most is entirely passive and so reporting rates are likely to underestimate true incidence. It is relatively efficient for rare, serious adverse effects and for those with a low underlying frequency in the population, but it is less useful when there is long period between administration and the event, or where the event has a relatively high underlying frequency. Greater emphasis should be placed on active surveillance after production registration. Detailed epidemiological investigations, including cohort, case control and cross-sectional designs, offer the only approaches that provide more information on the association between a product and events that have a high underlying frequency in the population or where there is a long period between administration and the adverse event. The relative merits of different approaches are discussed, with particular reference to our recently published study of the temporal association between canine vaccination and non specific signs of ill health and plans to undertake studies of associations with feline injection site sarcoma. Emphasis is placed on the need for clearly stated hypotheses and the consideration of equivalence, rather than significance testing when considering safety studies. PMID- 16713137 TI - Data collection on multiple births -- establishing twin registers and determining zygosity. AB - Twins are a valuable resource not only for studies on multiple births themselves, but on the etiology of diseases and other phenotypes. The method of ascertainment and selection of twins can be crucial for such studies and population based twin registries are thus of great importance as tools of research. Accurate determination of zygosity and chorionicity is essential in all studies of multiple births and in their professional care. The parents and the multiples ask for it. It is of pre-and postnatal medical importance and now considered as a prerequisite in several domains of twin research. It is also important for educational reasons as it helps the multiples and their parents and teachers to ascertain identity. The methods are briefly described and a plea is made to the obstetricians and pediatricians to use them systematically at birth. The distribution of zygosity and chorionicity types among spontaneous and induced twin births are illustrated. PMID- 16713139 TI - Relationships between innovative and traditional parameters to investigate semen quality in pigs. AB - Semen quality assessment represents a fundamental step for obtaining successful artificial insemination (AI). In commercial settings, the semen employed for AI should be of high quality but traditional semen quality estimates are not sufficiently sensitive to discriminate between differences among samples in terms of fertilising ability. Therefore, more discriminative sperm characteristics need to be identified in order to better predict fertility outcome. In the present study, a series of molecular aspects of semen, represented by heat shock proteins, oxidative stress status, antioxidant potential and tumour necrosis factor (TNF)-alpha were evaluated and analysed. Several relationships between traditional and investigated molecular semen quality estimates were found by using a multivariate analysis approach. Tumour necrosis factor (TNF)-alpha was identified in boar seminal plasma resulting in positive correlations with several sperm quality aspects and particularly with motility. The protective roles of antioxidant molecules and heat shock proteins have been demonstrated confirming the data previously published in the literature. PMID- 16713138 TI - Cigarette smoke condensate induces cytochromes P450 and aldo-keto reductases in oral cancer cells. AB - Our objective is to identify molecular factors which contribute to the increased risk of smokers for oral squamous cell carcinoma (OSCC). In the present study, we investigated the effects of cigarette smoke condensate (CSC) on gene expression profiles in different human oral cell phenotypes: normal epidermal keratinocytes (NHEK), oral dysplasia cell lines (Leuk1 and Leuk2), and a primary oral carcinoma cell line (101A). We determined differential gene expression patterns in CSC exposed versus non-exposed cells using high-density microarray RNA expression profiling and validation by quantitative real-time RT-PCR. A set of 35 genes was specifically up- or down-regulated following CSC treatment (25microg/ml for 24h) by at least 2-fold in any one cell type. Notably, five genes of the cytochrome P450 (CYP1A1, CYP1B1) and aldo-keto reductase (AKR1C1, AKR1C3, AKR1B10) families were highly increased in expression, some of them 15- to 30-fold. The timing and extent of induction for these genes differed among the four cell phenotypes. A potential biological interaction network for the CSC response in oral cells was derived from these data, proposing novel putative response pathways. These CSC responsive genes presumably participate in the prevention or repair of carcinogen induced DNA damage in tobacco-related oral carcinogenesis, and may potentially be exploited for determining the severity of exposure and for correcting mutagenic damage in exposed tissues of the oral cavity. PMID- 16713140 TI - Intensity of the biostimulatory effect of bulls on resumption of ovulatory activity in primiparous, suckled, beef cows. AB - Two experiments were conducted to evaluate postpartum resumption of ovulatory cycles in primiparous, suckled beef cows exposed to close physical contact or limited, fenceline contact with mature bulls. The hypotheses tested were that proportions of cows cycling and intervals from the start of exposure to resumption of ovulatory cycles did not differ among cows exposed to close physical contact; exposed to limited, fenceline contact; or not exposed to bulls. Angus x Hereford cows were assigned randomly to be exposed to close physical contact (experiment 1 (Exp. 1), BE; n = 14); limited, fenceline contact (Exp. 2, BEFL; n = 22); or not exposed (Exps. 1 and 2, NE; n = 37) to mature bulls beginning 59 days (Exp. 1) and 68 days (Exp. 2) after calving (day 0 in each experiment, respectively). In Exps. 1 and 2, anovular cows were exposed or not exposed to biostimulatory stimuli of bulls for 35 and 42 days, respectively. An increase in progesterone concentrations that exceeded 1.0 ng/mL in three consecutive samples was used as evidence of resumption of luteal function in Exp. 1. In Exp. 2, the presence of a corpus luteum in an ovary, assessed by ultrasonography, was used as evidence of resumption of luteal function. Percentage of cows that resumed luteal activity by the end of the Exp. 1 was greater (P < 0.05) for BE cows than for NE cows, whereas proportions of BEFL cows did not differ (P > 0.10) between BE or NE cows in Exp. 2. Interval from the start of the exposure period to resumption of luteal activity was shorter (P < 0.05) for BE and BEFL cows than for NE cows in Exps. 1 and 2, respectively. More (P < 0.05) BE than NE cows resumed luteal activity by 7 days after exposure and this difference was maintained throughout the exposure period in Exp. 1. However, in Exp. 2 proportions of BEFL cows that resumed luteal activity were greater (P < 0.05) than those of NE cows on days 14, 21, and 28 after exposure. By days 35 and 42, the proportion of BEFL cows that had resumed luteal activity did not differ (P > 0.10) from that NE cows. We conclude that the response of anovular, primiparous cows to the biostimulatory effect of bulls may depend on the intensity of exposure (frequency of exposure, duration of exposure, and quantity of stimuli) of pheromonal stimuli produced by bulls. PMID- 16713141 TI - A comprehensive proteomic analysis of the accessory sex gland fluid from mature Holstein bulls. AB - The expression of proteins in accessory sex gland fluid (AGF) of proven, high use mature Holstein bulls was evaluated. Thirty-seven bulls with documented fertility based on their non-return rates were studied. AGF was obtained by artificial vagina after bulls were surgically equipped with cannulae in the vasa deferentia. Samples of AGF were evaluated by two-dimensional SDS-PAGE, gels stained with Coomassie blue and polypeptide maps analyzed by PDQuest software. A master gel generated by the software representing the best pattern of spots in the AGF polypeptide maps was used as a reference for protein identification. Proteins were identified by Western blots and capillary liquid chromatography nanoelectrospray ionization tandem-mass spectrometry (CapLC-MS/MS). The product ion spectra were processed using Protein Lynx Global Server 2.1 prior to database search with both PLGS and MASCOT (Matrix Science) software. The entire NCBI database was considered for mass fingerprint matching. An average of 52+/-5 spots was detected in the AGF 2D gels, which corresponded to proteins potentially involved in capacitation (bovine seminal plasma protein-BSP-A1/A2 and A3, BSP 30 kDa, albumin); sperm membrane protection, prevention of oxidative stress, complement-mediated sperm destruction and anti-microbial activity (albumin, clusterin, acidic seminal fluid protein--aSFP, 5'-nucleotidase--5'-NT, phospholipase A2--PLA2); acrosome reaction and sperm-oocyte interaction (PLA2, osteopontin); interaction with the extracellular matrix (tissue inhibitor of metalloproteinase 2, clusterin) and sperm motility (aSFP, spermadhesin Z13, 5' NT). The 20 spots distinguished in all gels were matched to proteins associated with these functions. Proteins identified by tandem mass spectrometry as ecto-ADP ribosyltransferase 5 and nucleobindin, never described before in the accessory sex gland secretions, were also detected. In summary, we identified a diverse range of components in the accessory sex gland fluid of a select group of Holstein bulls with documented fertility. Known characteristics of these proteins suggest that they play important roles in sperm physiology after ejaculation. PMID- 16713142 TI - Bovine mural granulosa cells, and not the oocyte, are the major source of proteases capable of IGFBP-2 degradation. AB - Oocytes can regulate their own development, hence studies identifying and characterising oocyte-secreted factors are crucial to resolving the mechanisms by which oocytes can orchestrate follicular development. The insulin-like growth factor (IGF) system plays an important role during the development of a follicle; however, the regulation of IGF bioavailability is crucial throughout follicular growth. Proteolytic cleavage of the IGF/IGF binding protein (IGFBP) complex increases IGF bioavailability, hence, the production of IGFBP proteases by the oocyte and/or granulosa cells would provide a regulatory mechanism whereby they could regulate their own exposure to IGFs. The present study revealed mural granulosa cells (MGC), and not the oocyte, are the major source of proteases capable of cleaving IGFBP-2 in the developing bovine antral follicle. The addition of recombinant IGF-I or FSH had no effect in terms of modulating IGFBP-2 degradation. This work further supports the existence of a local regulatory mechanism modulating IGF bioavailability. PMID- 16713143 TI - Conventional-dose hormone therapy (HT) and tibolone, but not low-dose HT and raloxifene, increase markers of activated coagulation. AB - OBJECTIVES: Hormone therapy (HT) is associated with a modest, but significantly increased risk for arterial and venous thromboembolism. We have compared the effects of estrogen, tibolone, and raloxifene on relevant markers of coagulation activation and investigated whether there is a dose-response relationship of oral HT. METHODS: Randomized, open-label, comparative study of 202 healthy women who were assigned to receive treatment for 12 weeks with either low-dose hormone therapy containing 1 mg 17beta-estradiol + 0.5 mg norethisterone acetate (NETA) (n=50), conventional-dose HT containing 2 mg 17beta-estradiol and 1 mg NETA (n=50), 2.5 mg tibolone (n=51), or 60 mg raloxifene (n=51). RESULTS: The groups were comparable with regard to demographic characteristics and laboratory variables at baseline. D-dimer increased markedly in the conventional-dose HT group, but remained unchanged in the low-dose HT group. Tibolone was associated with a medium increase, whereas raloxifene was associated with a decrease in D dimer levels. Changes in prothrombin fragment 1 + 2 showed a similar pattern for all four groups, whereas no significant differences in changes of thrombin antithrombin complex were observed. CONCLUSIONS: Our data suggest that low-dose HT is associated with less activation of coagulation than conventional-dose HT. This finding may be of clinical importance since randomized clinical trials showing increased risk of thrombosis have utilized conventional-dose HT. PMID- 16713144 TI - The use of Stokes deformation number as a predictive tool for material exchange behaviour of granules in the 'equilibrium phase' in high shear granulation. AB - The objective of this study was three-fold; to investigate the different mechanisms of material exchange during the equilibrium phase of the granulation process and whether these mechanisms are consistent with the mechanisms described in the growth regime map, to study how material properties and process conditions affect these exchange mechanisms, and to correlate Stokes deformation number to the exchange mechanisms. Microcrystalline cellulose (MCC), alpha-lactose, microfine cellulose (MFC), and dextrin were granulated using water as a binding agent. Once in the equilibrium phase, 5% (w/w) of the granular mass was replaced with wet tracer granules, after which the granulation process was continued. Granules were typically of a size of approximately 1mm in diameter. Therefore, these granules can also be called pellets. Tracer experiments show indeed solid material exchange can take place in the equilibrium phase of the high shear granulation process. Tracer material was equally dispersed throughout the whole batch for all materials tested. However, the granulation time needed to reach this homogeneous distribution varied with material and granulation conditions. Three different mechanisms of material exchange were identified: exchange by disintegration, where granules are rapidly crushed and formed to granules again; exchange by deformation, where abraded granule fragments immediately fuse with other granules; and exchange by distribution, where there is a prolonged period over which both tracer and standard granules stay intact, followed by uncontrolled growth and exchange of material. It was found that it is possible to shift between the mechanisms by changing the process conditions, e.g., changing viscosity or amount of binder liquid. These observations indicate that by choosing the appropriate process conditions improved distribution of small amounts of insoluble materials in the granules can be obtained. A relation exists between the exchange mechanisms and the growth regime map: the disintegration mechanism resembles 'crumb behaviour', the deformation mechanism resembles 'steady growth', and the distribution mechanism resembles 'nucleation' and 'induction growth'. Unfortunately, Stokes deformation number cannot be used as a predictive tool when low viscosity binders like water are used, due to the importance of viscosity in the equation. However, this number is one of the variables of the growth regime map. Since the exchange mechanisms correspond to the granule growth mechanisms in the regime map, alternatively colour experiments might be used to reveal the granulation regime. PMID- 16713145 TI - Encapsulation in lipospheres of the complex between butyl methoxydibenzoylmethane and hydroxypropyl-beta-cyclodextrin. AB - The aim of this study was to investigate the incorporation into lipospheres of the complex between hydroxypropyl-beta-cyclodextrin (HP-beta-CD) and the sunscreen agent, butyl methoxydibenzoylmethane (BMDBM) and to examine the influence of this system on the sunscreen photostability. The formation of the inclusion complex was confirmed by thermal analysis and powder X-ray diffraction. Lipid microparticles loaded with free BMDBM or its complex with HP-beta-CD were prepared using tristearin as the lipid material and hydrogenated phosphatidylcholine as the emulsifier. The obtained lipospheres were characterized by scanning electron microscopy and differential scanning calorimetry. The microparticle size (15-40 microm) was not affected by the presence of the complex. Release of BMDBM from the lipospheres was lower when it was incorporated as inclusion complex rather than as free molecule. Unencapsulated BMDBM, its complex with HP-beta-CD, the sunscreen-loaded lipospheres or the lipoparticles containing the BMDBM/HP-beta-CD complex, were introduced into a model cream (oil-in-water emulsion) and irradiated with a solar simulator. The photodegradation studies showed that all the examined systems achieved a significant reduction of the light-induced decomposition of the free sunscreen agent (the BMDBM loss decreased from 28.9 to 17.3-15.2%). However, photolysis experiments performed during 3 months storage of the formulations, demonstrated that the photoprotective properties of the HP-beta-CD complex and of BMDBM alone-loaded lipospheres decreased over time, whereas the microencapsulated HP-beta-CD/BMDBM complex retained its photostabilization efficacy. Therefore, incorporation in lipid microparticles of BMDBM in the cyclodextrin complex form is more effective in enhancing the sunscreen photostability than the complex alone or the liposphere-entrapped free BMDBM. PMID- 16713146 TI - Inhibition of malarial topoisomerase II in Plasmodium falciparum by antisense nanoparticles. AB - New effective antimalarial agents are urgently needed due to increasing drug resistance of Plasmodium falciparum. Phosphorothioate antisense oligodeoxynucleotides (ODNs) silencing of malarial topoisomerase II gene have shown to possess promising features as anti malarial agents. In order to improve stability and to increase intracellular penetration, ODNs were complexed with the biodegradable polymer chitosan to form solid nanoparticles with an initial diameter of approximately 55 nm. The particle zetapotential depended on the chitosan/ODN mass ratio. Nanoparticles with mass ratio of 2:1 displayed a positive surface charge (+15 mV) whereas particles with 1:1 mass ratio were negatively charged (-20 mV). Additionally nanoparticles were found to protect ODNs from nuclease degradation. P. falciparum K1 strain was exposed to the chitosan/ODN-nanoparticles for 48 h in order to examine the effects of chitosan/antisense (AS) and chitosan/sense (S) oligodeoxynucleotide nanoparticles on malaria parasite growth. Both negatively and positively charged antisense nanoparticles as well as free antisense ODNs (in a final concentration of 0.5 microM) showed sequence specific inhibition compared with sense sequence controls. However, nanoparticles were much more sequence specific in their antisense effect than free ODNs. Nanoparticles with negative surface charge exhibited a significantly stronger inhibitory effect ( approximately 87% inhibition) on the parasite growth in comparison to the positive ones ( approximately 74% inhibition) or free ODNs ( approximately 68% inhibition). This is the first study demonstrating the susceptibility of P. falciparum to antisense nanoparticles. PMID- 16713147 TI - PEGylated J591 mAb loaded in PLGA-PEG-PLGA tri-block copolymer for targeted delivery: in vitro evaluation in human prostate cancer cells. AB - J591 monoclonal antibody (mAb) has high affinity for prostate specific membrane antigen (PSMA) on prostate cancer (PCA) cells. We coupled polyethylene glycol J591 (PEGylated J591) to a salicyl hydroxamic acid (SHA)-derivatized polyethylenimine (PEI)/DNA-betagal vector to investigate the specificity and efficiency of targeting PSMA in PCA cells through encapsulation. Coupling was facilitated via the high affinity interaction between phenyl(di)boronic acid (PDBA) and SHA molecules yielding J591/PEG/PEI/DNA-betagal polyplex. After encapsulation with poly(d,l-lactic-co-glycolic acid)-b-polyethylene glycol-b poly(d,l-lactic-co-glycolic acid) (PLGA-PEG-PLGA) tri-block copolymer, 8-10-fold increment of gene transfection levels were attained at the optimum concentration of 0.25% (w/v) using Pluronic F68 tri-block copolymer as a control. The enhanced transfection efficiency was attributed to increased internalization and uptake of the radiolabeled plasmid in the presence of PLGA-PEG-PLGA tri-block copolymer. The release of plasmid DNA (pDNA) from microparticles containing SHA-PEI complexed pDNA showed little initial burst release followed by a 5% release over 48 h. The release accelerated thereafter and approximately 60% was released after 28 days. Deconvolution confocal microscopy showed polyplex/microparticle formulation localized in the cell nucleus as opposed to the polyplex without PLGA PEG-PLGA indicating that an optimal concentration of PLGA-PEG-PLGA tri-block copolymer can be utilized to enhance endocytic process of J591-mediated targeting of PCA cells. PMID- 16713148 TI - Susceptibility of nanoparticle-encapsulated paclitaxel to P-glycoprotein-mediated drug efflux. AB - Overexpression of P-glycoprotein (P-gp) is a key factor contributing to the development of multidrug resistance (MDR) in cancer cells. The objective of the study is to investigate whether a P-gp substrate, paclitaxel, delivered to MDR tumor cells in poly(d,l-lactide-co-glycolide) (PLGA) nanoparticles is susceptible to P-gp - mediated drug efflux. Paclitaxel-loaded nanoparticles were formulated by emulsion-solvent evaporation technique. Nanoparticles had a mean hydrodynamic diameter of about 195nm, and demonstrated sustained release of paclitaxel. In vitro cell culture studies indicated that paclitaxel nanoparticles result in sustained, dose-dependent and significant cytotoxicity in drug-sensitive MCF-7 tumor cells but not in drug-resistant NCI-ADR/RES cells. Resistance to nanoparticle-encapsulated paclitaxel was reversed by verapamil, a P-gp inhibitor. Further, sustained inhibition of P-gp was necessary for sustaining the cytotoxicity of nanoparticle-encapsulated paclitaxel in drug-resistant cells. Inhibition of P-gp by verapamil did not significantly affect the uptake or retention of nanoparticles in drug-resistant cells. In conclusion, our studies suggest that P-gp substrates, such as paclitaxel, delivered to MDR cells by PLGA nanoparticles, are susceptible to efflux by P-gp. Inhibition of P-gp restores sensitivity to paclitaxel; however, sustained inhibition of P-gp is required for sustained therapeutic efficacy of nanoparticle-encapsulated drug. PMID- 16713149 TI - Biodistribution and tumor-accumulation of gadolinium (Gd) encapsulated in long circulating liposomes in tumor-bearing mice for potential neutron capture therapy. AB - To deliver and maintain a sufficient amount of Gd into tumors is required for a successful Gd neutron capture therapy (Gd-NCT), but it has been proven to be rather challenging to achieve. Previously, we have reported a Gd-encapsulated liposome formulation that has the potential to overcome this challenge. In the present study, we sought to systemically evaluate the biodistribution and the tumor-accumulation of the Gd in model tumor-bearing mice. The Gd-encapsulated liposomes were injected into mice pre-grafted with two different model tumors. The Gd content in the tumors and other organs were determined at various time after the injection. A sufficient amount of Gd was readily delivered into those two different model tumors. Increasing the dose of Gd by injecting the Gd encapsulated liposomes multiple times tended to increase the uptake of the Gd by the tumors. Finally, the uptake of Gd by tumors was inversely correlated with the size of the tumors. The Gd-encapsulated liposomes hold great potentials as a Gd delivery system for NCT of small- and medium-size tumors. Alternative strategies may have to be adopted in order to use NCT to treat large, advanced solid tumors, although for which, Gd-NCT might be advantageous over boron-NCT. PMID- 16713150 TI - Formulation and cytotoxicity of doxorubicin nanoparticles carried by dry powder aerosol particles. AB - Regional drug delivery via dry powder inhalers offers many advantages in the management of pharmaceutical compounds for the prevention and treatment of respiratory diseases. In the present study, doxorubicin (DOX)-loaded nanoparticles were incorporated as colloidal drug delivery system into inhalable carrier particles using a spray-freeze-drying technique. The cytotoxic effects of free DOX, carrier particles containing blank nanoparticles or DOX-loaded nanoparticles on H460 and A549 lung cancer cells were assessed using a colorimetric XTT cell viability assay. The mean geometric carrier particle size of 10+/-4 microm was determined using confocal laser scanning microscopy. DOX loaded nanoparticles had a particle size of 173+/-43 nm after re-dissolving of the carrier particles. Compared to H460 cells, A549 cells showed less sensitivity to the treatment with free DOX. The DOX-nanoparticles showed in both cell lines a higher cytotoxicity at the highest tested concentration compared to the blank nanoparticles and the free DOX. The cell uptake of free DOX and DOX delivered by nanoparticles was confirmed using confocal laser scanning microscopy. This study supports the approach of lung cancer treatment using nanoparticles in dry powder aerosol form. PMID- 16713151 TI - Evaluation of the interaction of surfactants with stratum corneum model membrane from Bothrops jararaca by DSC. AB - The interaction of surfactants sodium dodecyl sulfate (SDS), cetyl trimethyl ammonium chloride (CTAC) and lauryl alcohol ethoxylated (12 mol ethylene oxide) (LAE-12OE) was evaluated on the stratum corneum (SC) of shed snake skins from Bothrops jararaca, used as model membrane, and thermal characterized by differential scanning calorimetry (DSC). Surfactant solutions were employed above of the critical micellar concentration (CMC) with treatment time of 8h. The SDS interaction with the SC model membrane has increased the characteristic transition temperature of 130 degrees C in approximately 10 degrees C for the water loss and keratin denaturation, indicating an augmentation of the water content. Samples treated with CTAC have a decrease of the water loss temperature, while, for the LAE-12OE treated samples, changes on the transition temperature have not been observed. PMID- 16713152 TI - Retinol-encapsulated low molecular water-soluble chitosan nanoparticles. AB - This aim of this study was to encapsulate retinol into chitosan nanoparticles and reconstitute it into aqueous solution. Retinol-encapsulated chitosan nanoparticles were prepared for application of cosmetic and pharmaceutical applications. Retinol-encapsulated chitosan nanoparticle has a spherical shape and its particle sizes were around 50-200 nm according to the drug contents. Particle size was increased according to the increase of drug contents. Solubility of retinol is able to increase by encapsulation into chitosan nanoparticles more than 1600-fold. It was suggested that retinol was encapsulated into chitosan nanoparticles by ion complex as a result of FT-IR spectra. Specific peak of chitosan at 1590 cm(-1) was divided to semi-doublet due to the electrostatic interaction between amine group of chitosan and hydroxyl group of retinol. At (1)H NMR spectra, specific peaks of retinol disappeared when retinol encapsulated chitosan nanoparticles were reconstituted into D(2)O while specific peaks both of retinol and chitosan appeared at D(2)O/DMSO (1/4, v/v) mixture. XRD patterns also showed that crystal peaks of retinol were disappeared by encapsulation into chitosan nanoparticles. Retinol-encapsulated nanoparticles were completely reconstituted into aqueous solution as same as original aqueous solution and zeta potential of reconstituted chitosan nanoparticles was similar to their original solution. At HPLC study, retinol was stably and efficiently encapsulated into chitosan nanoparticles. PMID- 16713153 TI - Constituents and biological activities of Schinus polygamus. AB - The folk medicine employs Schinus polygamus to treat arthritic pain and cleansing of wounds. As no reports of pharmacological studies supporting its anti inflammatory and analgesic properties, extracts of increasing polarity were assayed on the base of fever, pain and inflammation, together with its antimicrobial activity. All the extracts showed pharmacological activities. From the most active extracts different metabolites were isolated that can in part explain the antipyretic, anti-inflammatory, and analgesic activity: beta sitosterol, shikimic acid together with quercetin, previously reported. Also, the essential oil of leaves and fruits was obtained and compared with the oil obtained from Schinus polygamus collected in Argentine. Oils differed in composition and in antibacterial activity, where the Chilean species exhibited a wide spectrum of activity against Gram-positive and Gram-negative bacteria, and the most abundant compound found in leaves and fruits was beta-pinene, meanwhile the Argentine species showed high activity against Bacillus cereus, and the main components resulted to be alpha-phellandrene and limonene. PMID- 16713154 TI - Antimutagenic activity of methanolic extract of Ganoderma lucidum and its effect on hepatic damage caused by benzo[a]pyrene. AB - The antimutagenic activity of the methanolic extract of the fruiting bodies of Ganoderma lucidum (Fr.) P. Krast. occurring in South India was investigated. The activity was assayed by Ames Salmonella mutagenicity test using histidine mutants of Salmonella typhimurium tester strains, TA98, TA100 and TA102. The methanolic extract of the mushroom significantly inhibited (P<0.001) the in vitro sodium azide (NaN(3)), N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 4-nitro-o phenylenediamine (NPD), and benzo[a]pyrene (B[a]P) induced his(+) revertants in a dose dependent manner. In vivo antimutagenic activity of extract was also assayed by determining the mutagenicity of the urine of rats administrated with B[a]P as a mutagen. The prior administration of extract markedly inhibited mutagenicity induced by B[a]P. The results indicated that the methanolic extract of Ganoderma lucidum occurring in South India possessed significant antimutagenic activity. The effect of B[a]P on hepatic enzymes, such as serum glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT) and alkaline phosphtase (ALP), were also evaluated. The extract prevented the increase of SGOT, SGPT, and ALP activities consequent to B[a]P challenge, and enhanced the levels of reduced glutathione (GSH) and activities of glutathione peroxidase (GPx), glutathione-S transferase (GST), superoxide dismutase (SOD), and catalase (CAT). The extract also profoundly inhibited lipid peroxidation induced by B[a]P. The results revealed that Ganoderma lucidum extract restored antioxidant defense and prevented hepatic damage consequent to the challenge by B[a]P. PMID- 16713155 TI - Inhibition of intestinal motility and secretion by extracts of Epilobium spp. in mice. AB - Ethanol extracts of the fresh aerial parts of various Epilobium species were tested to elucidate the mechanism of their gastrointestinal activity in animals. The methods of charcoal meal, castor oil-induced diarrhoea, and enteropooling assay were used to evaluate their effect on mouse gut at various dose levels. The extracts were found to have a significant activity in all models. Moreover, the extracts resulted to possess very little toxicity. Thus, it can be concluded that Epilobium possesses anti-diarrhoeal, anti-motility, and anti-secretory activities and can prove beneficial in the treatment of gastrointestinal disorders. PMID- 16713156 TI - Antihypercholesterolaemic and antioxidant activity assessment of some plants used as remedy in Turkish folk medicine. AB - Ethanolic and aqueous extracts from five plant species used in Turkish traditional medicine were evaluated for in vivo hypercholesterolaemic and antioxidant activities: Agrostemma githago L., Potentilla reptans L., Thymbra spicata var. spicata L., Urtica dioica L. and Viscum album var. album L. We assayed the effects of the administration of plant extracts on serum total cholesterol, triglyceride, HDL-C, LDL-C, glucose, AST and ALT concentrations in mice fed with cholesterol-rich diet. In addition, plasma TAA, MDA and NO(x) levels in the same animals were assayed. All the aqueous plant extracts did not affect the serum cholesterol concentration. However, the ethanolic extracts of Agrostemma githago, Thymbra spicata and Viscum album decreased the serum cholesterol concentration in the mice fed with high-cholesterol diet without inducing any gastric damage. The ethanolic extracts of Thymbra spicata, Viscum album, Potentilla reptans and Urtica dioica and the aqueous extract of Agrostemma githago increased the serum HDL concentration, whereas the ethanolic extracts of Agrostemma githago, Thymbra spicata, Viscum album and Urtica dioica decreased the serum LDL-C concentration. Thymbra spicata and Viscum album were observed to decrease the serum triglyceride concentration. Among the plant extracts studied, the ethanolic extracts of Thymbra spicata significantly decreased the MDA level in mice. The ethanolic extract of Potentilla reptans increased in NO(x). None of these plants showed statistically prominent activity on plasma TAA. Results of the present study indicated that the ethanolic extracts of Agrostemma githago, Thymbra spicata and Viscum album showed potent hypocholesterolaemic activity in the mice fed with a diet containing high-cholesterol. PMID- 16713157 TI - Antimalarial activity of some Colombian medicinal plants. AB - Antimalarial activity of 10 vegetal extracts (9 ethanolic extracts and 1 crude alkaloid extract), obtained from eight species traditionally used in Colombia to treat malaria symptoms, was evaluated in culture using Plasmodium falciparum chloroquine resistant (FcB2) strain and in vivo on rodent malaria Plasmodium berghei. The activity on ferriprotoporphyrin biomineralization inhibition test (FBIT) was also assessed. Against Plasmodium falciparum, eight extracts displayed good activity Abuta grandifolia (Mart.) Sandwith (Menispermaceae) leaves, Acacia farnesiana (L.) Willd. (Mimosaceae) leaves, Acnistus arborescens (L.) Schltdl. (Solanaceae) aerial part, Croton leptostachyus Kunth (Euphorbiaceae) aerial part, Piper cumanense Kunth (Piperaceae) fruits and leaves, Piper holtonii C. DC. (Piperaceae) aerial part and Xylopia aromatica (Lam.) Mart. (Annonaceae) bark with IC(50) values ranging from <1 to 2.1 microg/ml, while in the in vivo model only Abuta grandifolia alkaloid crude extract exhibits activity, inhibiting 66% of the parasite growth at 250 mg/kg/day. In the FBIT model, five extracts were active (Abuta grandifolia, Croton leptostachyus, Piper cumanense fruit and leaves and Xylopia aromatica). PMID- 16713158 TI - Some unique ethnomedicinal uses of plants used by the Korku tribe of Amravati district of Maharashtra, India. AB - The present ethnobotanical study was carried out among the ethnic groups (Korku tribe) in the Chikhaldara, Achalpur and parts of Morshi in Amravati district of Maharashtra, India. A field survey of the study area was carried out during 2000 2004 to document the medicinal utility of plants occurring in this area by Korku tribe. Traditional uses of 66 plant species belonging to 40 families are described under this study. The documented ethnomedicinal plants were mostly used to cure skin disorders, diarrhea, jaundice, tuberculosis, stroke, migraine, menstrual problems, fertility problems, urinary problems, piles, wounds and poison bites. The medicinal plants used by the Korku tribe are arranged alphabetically followed by family name, herbarium number, local name, parts used, mode of preparation and medicinal uses. PMID- 16713159 TI - Study on the antinociceptive effects of Thymus broussonetii Boiss extracts in mice and rats. AB - In Morocco, Thymus broussonetii is widely used in folk medicine for the treatment of a variety of diseases including gastroenteric and bronchopulmonary disorders and to relieve dolorous process. The antinociceptive effect of the aqueous, butanolic and ethyl acetate extracts of this species was examined in rats and mice using chemical and thermal models. The results obtained showed that aqueous and butanolic extracts exerted an antinociceptive activity in the two phases of formalin (50-300 mg/kg), tail immersion and writhing tests. Whereas, the ethyl acetate extract reduced the nociceptive response only in the second phase of formalin (100-300 mg/kg) and writhing tests. The aqueous extract, which is the most effective, contains active analgesic principles acting both centrally and peripherally. Furthermore, this antinociceptive effect has been avoided by naloxone at a dose of 1mg/kg in the first phase of formalin and hot plate tests indicating that this extract acts partly through an opioid-mediated mechanism. The present results demonstrated that Thymus broussonetii contains active constituents which possess antinociceptive activity justifying its popular use to relieve some pains. PMID- 16713160 TI - Studies on the luteolytic, oestrogenic and follicle-stimulating hormone like activity of plant Rhaphidophora pertusa (Roxb.). AB - The phytochemical, pharmacological and toxicological feature of plant Rhaphidophora pertusa (Roxb.) was done. Phytosteroids, flavonoids, tannins and glucosides were detected in the plant extracts. In cross-bred (Zebu X Holstein Friesian or Jersey) dairy cows, subsequent to prostaglandin (PG) induced oestrus, to each group (n=4), cloprostenol (PG control) 100 microg i.m. on day 10, the rice gruel (vehicle) was fed on day 10 or the fresh stem (1 kg/animal/day) in rice gruel on day 9, or days 9 and 10, or days 9-11 of the oestrous cycle. Each group received subcutaneously either 5% gum acacia suspension or the plant ethyl acetate or methanol extract (1g in gum acacia) on days 8 (to bannur ewes) or 10 (to dairy cows) of the oestrous cycle. In PG control cows or ewes, there was induction of oestrus in 48 h and a fall in serum progesterone concentration. The feeding of fresh stem in the rice gruel or the s.c. administration of the plant extract did not induce oestrus or significantly (P>0.05) alter the serum progesterone, bilirubin, calcium, creatinine, phosphorus, magnesium and glucose concentrations or the total erythrocyte and leucocyte count, differential leucocyte count and haemoglobin concentration. The plant did not cause any toxicity in the cow or ewe. In immature rats, the aqueous or methanol (hot or cold) extract did not cause any follicle-stimulating hormone (FSH)-like activity. The methanol extract increased the uterine weight in ovariectomised rats. This suggested the presence of oestrogenic activity in the plant. In conclusion, the present study revealed the presence of oestrogenic activity in the plant and the absence of luteolytic or FSH-like or toxic activity. PMID- 16713161 TI - Basal ganglia lesions in a patient with 3-hydroxyisobutyric aciduria. AB - 3-hydroxyisobutyric aciduria (3HiB-uria) is a very rare organic aciduria that involves valine metabolism. We report the case of a 7-year-old boy with 3HiB-uria who has suffered more than 20 ketoacidotic episodes since the age of 15 months. In the most recent ketoacidotic episode, which was particularly severe, he developed mild dystonia and choreoathetosis. Magnetic resonance imaging (MRI) revealed bilateral swelling and signal abnormalities of the putamina and heads of the caudate nuclei. The abnormal movements showed a gradual improvement over several months, in correlation with neuroradiological findings. 3HiB-uria should be recognized as one of the group of branched chain organic acidemias that can produce lesions in the basal ganglia. PMID- 16713162 TI - [Hemostasis disorders in severe infections: state of the art]. AB - The haemostatic system is greatly modified during severe infections. The early activation of coagulation is triggered by tissue factor expression and secondary fibrinolysis impaired by the upregulation of fibrinolysis inhibitors. This imbalance is a major cause of subsequent organ dysfunction. Natural anticoagulants (Tissue factor pathway inhibitor (TFPI), Antithrombin (AT), and Protein C (PC) are consumed or inhibited in this pathological process justifying a therapeutic supplementation with these inhibitors to improve sepsis-induced organ failure and mortality. No effect on the mortality rate could be documented in controlled studies using recombinant TFPI or AT concentrates but a biological interaction with heparin therapy could have biased the results. Treatment with recombinant activated PC was associated with a significant reduction in the mortality rate of severely ill patients. An increase in the rate of hemorrhagic adverse effects was observed with these compounds, justifying a strict observance of contraindications and of patient selection. PMID- 16713164 TI - Rapid detection of rifampicin-resistant Mycobacterium tuberculosis by in-house, reverse line blot assay. AB - Drug resistance in tuberculosis is a significant problem in countries endemic for tuberculosis. A sensitive, specific, and high-throughput reverse line blot assay (RLBA) for the detection of genotypic resistance to rifampicin (RIF) was designed and evaluated. DNA sequencing done for 72 resistant isolates from Delhi, for baseline data, showed mutations within the rpoB core region in all RIF-resistant strains. The RLBA includes oligonucleotide probes specific for wild-type and mutant sequences, allowing sensitive detection of both genotypes in a single assay. The assay based on reverse hybridization principle simultaneously detects 13 different mutations affecting 6 independent codons, including the most prevalent mutations at positions 531 and 526. Application of the method to a panel of 292 MDR TB isolates and susceptible strains from 5 different cities in India showed 98% concordance with the sequencing results. This rapid, simple, economical, and highly sensitive assay provides a practical alternative to sequencing for genotypic evaluation of RIF resistance in Mycobacterium tuberculosis. PMID- 16713163 TI - Kinetic analysis of the humoral immune response against 3 Toxoplasma gondii recombinant proteins in infants with suspected congenital toxoplasmosis. AB - The reactivity values of Toxoplasma gondii ROP2, GRA4, and GRA7 recombinant antigens (rAgs) were analyzed by immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) in 23 congenitally infected (I) and 36 noninfected (NI) infants. The reactivity values observed from the serum samples of I versus NI infants for each recombinant protein were 91% versus 67% for rROP2 (P = .05), 86% versus 29% for rGRA4 (P < .001), and 56% versus 11% for rGRA7 (P = .003). The follow-up showed that serum samples from NI infants became negative for specific IgG at 5.8 months (95% confidence interval [CI], 4.9-6.7) using a commercial assay; meanwhile, by specific recombinant protein ELISA, the samples became negative at 3.7 months with rROP2 (95% CI, 2.8-4.6), at 1.3 months with rGRA4 (95% CI, 0.8-1.8), and at 0.9 months with rGRA7 (95% CI, 0.5-1.3). Kinetic analysis also showed that serum samples from group I presented different IgG profiles among rAgs. The rROP2 IgG profile was similar to that of the commercial assay, whereas rGRA4 and rGRA7 profiles showed a gradual decrease along the period of the study. The potential of the utility of rAgs to develop a diagnostic system that discriminates congenitally I infants from NI is discussed. PMID- 16713165 TI - The microbiology of bacteriuria in men: a 5-year study at a Veterans' Affairs hospital. AB - Bacteriuria and urinary tract infection occur relatively frequently in older men, but data regarding the causative microorganisms are limited. We retrospectively identified all positive cultures of urine specimens (n = 4943) obtained over a 5 year period at our institution. We determined the frequency of causative microorganisms and grouped these by Gram type, setting of patient care, and method of urine specimen collection. We also assessed the performance characteristics of the Gram-stained smear of uncentrifuged urine. Among our patients, Gram-positive cocci (GPC) were isolated as often as Gram-negative rods (GNR). Escherichia coli was the single or predominant isolate in only 14% of cases, and Enterococcus was the single most commonly identified genus (22.5%). The Gram stain was accurate in predicting the culture results (positive likelihood ratio, 7.0 for GPC and 8.1 for GNR). We conclude that the microorganisms causing bacteriuria in older male veterans are substantially different from those found in women, and the Gram-stained smear provides useful information on the causative organisms. PMID- 16713166 TI - Effects of progestagens on follicular growth and oocyte developmental competence in FSH-treated ewes. AB - Previous research has reported evidence for negative effects of progestagens on follicular growth and oocyte competence. In the present study, negative effects of progestagens on follicular growth and oocyte developmental competence were assessed. During the breeding season, 20 Sarda ewes were treated with two doses of cloprostenol, 10 days apart, to assure the presence of a corpus luteum (CL). On day 5 after the second cloprostenol dose, 10 ewes were treated with a progestagen sponge while 10 females remained untreated. Starting on day 7 after the second cloprostenol dose, all the ewes were treated with 6 equal doses of 24 I.U. of FSH (Ovagen, ICP, NZ), every 12h. The number of follicles > or =2mm in diameter increased (P<0.0005) in all the ewes from 24 h before to 60 h after the first FSH dose (from 12.8+/-1.1 to 23.4+/-1.3 in treated and from 12+/-0.6 to 22+/-1.2 in untreated ewes, n.s.). There were no significant differences in follicle dynamics between groups, but concentrations of estradiol in control ewes were higher than in the progestagen group (P<0.05). Twelve hours after the last FSH dose, oocytes were collected by ovum pick-up. Recovery rates were lower for progestagen-treated ewes (71.1 versus 83%; P<0.001). After IVP procedure, cleavage rate was also lower in the progestagen group (39.1 versus 82.6%; P<0.001). Furthermore, blastocysts output revealed that oocyte developmental competence was lower in progestagen group (17.3 versus 30.4%; P=0.245), although differences were not significant. These results suggest deleterious effects from progestagen on oocyte developmental competence and set the basis for new protocols for in vitro embryo production. PMID- 16713167 TI - [Cerebral microdialysis as a clinical tool]. AB - Microdialysis is the only technique available for cerebral metabolic monitoring in the clinical setting. By the mean of a probe inserted in the brain, it provides an extracellular space sampling. Values of various substrates including cerebral glucose, lactate, pyruvate, glycerol or glutamate can be obtained at the bedside at intervals between minutes and hours. Values are critically dependent on the flow of the perfusion liquid and reflect a highly localized cerebral metabolism. Cerebral microdialysis improves our understanding of acute neurological events such as intracranial hypertension or decrease in brain tissue oxygen pressure. Cerebral microdialysis can be used for detection of ischaemia, especially after malignant stroke or vasospasm complicating subarachnoid haemorrhage. In these cases, it may influence the therapeutic management. Moreover, it permits the assessment of metabolic changes after therapeutic interventions. Finally, some markers (like lactate/pyruvate ratio) are related to outcome, especially after traumatic brain injury. PMID- 16713168 TI - [What sedation for prevention and treatment secondary brain insult?]. AB - The aim of sedation and analgesia is to prevent secondary brain insult. The goals of sedation are the prevention and treatment of intracranial hypertension and systemic disorders. In such situation, the use of sedative and analgesic therapy should respect the rate of cerebral blood flow/cerebral oxygen consumption coupling while preserving cerebral perfusion pressure and decreasing the intracranial pressure. This treatment should have an analgesic and myorelaxing action with short and predictable time of action. The optimal agent with all these characteristics does not exist, but the combination of several pharmacological compounds may reach this goal. Benzodiazepines are the most frequently agents used. In most of cases they are associated with analgesics like opioids or ketamine. Opioids are the basis of analgesia because they do not produce brain haemodynamic alterations if arterial pressure is maintained. Ketamine, which use in this indication is matter of debate, has the advantage to maintain haemodynamics. Ketamine has no side effects on brain haemodynamics when used in combination with propofol or midazolam. Because of their side effects on haemodynamics and immune response, barbituric are no longer used as long term sedative agents. However, they are still recommended in cases of refractory intracranial hypertension. Propofol remains the optimal sedative agent because of its short duration action although its use is restricted because it is an expensive drug. Its use is recommended for short time sedation with or without opioids. The use of neuromuscular blockers should be focused on the patients with an intracranial hypertension refractory to standard treatment. The presence of brain damage in patients makes difficult to assess the level of sedation. One should avoid over sedation, which increases morbidity by prolongation of the duration of mechanical ventilation. PMID- 16713169 TI - Application of a population balance model to a perfusion in vitro toxicity system. AB - Perfusion cell culture is different to static cell culture: In perfusion culture, steady state concentrations of nutrients and metabolites can be achieved. When a toxin is added to the medium, it is administered to the cells at a defined constant concentration. Perfusion culture is favourable for time series analysis of experimental data because assays can be performed online in the outflowing medium at a high time resolution. The cytotoxicity exerted by cadmium chloride (CdCl(2)) on LLC-PK(1) cells was investigated in static and in perfusion culture using the EpiFlow system in order to compare both the techniques. For this comparison, cytotoxicity was assessed by the release of the cytosolic enzyme lactate dehydrogenase (LDH), and by the potential of viable cells to reduce resazurin. The results showed a greater sensitivity of LLC-PK(1) monolayers to CdCl(2) with continuous exposure in perfusion culture as compared to a single or repeated dose in static culture, as evidenced by increased LDH release and decreased resazurin reduction. These differences between static and perfusion culture were ascribed to different delivery rates of toxin to the site of absorption. Cadmium accumulation of the cells was measured and found to be higher under perfusion conditions than under static conditions. Time series analysis of the LDH release in perfusion culture was performed by establishing a deterministic population balance model of cell death. An empirical death rate function that accounts for accumulation of the toxin was introduced and could be successfully verified. PMID- 16713170 TI - Correlation of serpin-protease expression by comparative analysis of real-time PCR profiling data. AB - Imbalanced protease activity has long been recognized in the progression of disease states such as cancer and inflammation. Serpins, the largest family of endogenous protease inhibitors, target a wide variety of serine and cysteine proteases and play a role in a number of physiological and pathological states. The expression profiles of 20 serpins and 105 serine and cysteine proteases were determined across a panel of normal and diseased human tissues. In general, expression of serpins was highly restricted in both normal and diseased tissues, suggesting defined physiological roles for these protease inhibitors. A high correlation in expression for a particular serpin-protease pair in healthy tissues was often predictive of a biological interaction. The most striking finding was the dramatic change observed in the regulation of expression between proteases and their cognate inhibitors in diseased tissues. The loss of regulated serpin-protease matched expression may underlie the imbalanced protease activity observed in pathological states. PMID- 16713171 TI - Analysis of copy number variation in the normal human population within a region containing complex segmental duplications on 22q11 using high-resolution array CGH. AB - A previously detected copy number polymorphism (Ep CNP) in patients affected with neuroectodermal tumors led us to investigate its frequency and length in the normal population. For this purpose, a program called Sequence Allocator was developed and applied for the construction of an array that consisted of unique and duplicated fragments, allowing the assessment of copy number variation within regions of segmental duplications. The average resolution of this array was 11 kb and we determined the size of the Ep CNP to be 290 kb. Analysis of normal controls identified 7.7 and 7.1% gains in peripheral blood and lymphoblastoid cell line (LCL) DNA, respectively, while deletions were found only in the LCL group (7.1%). This array platform allows the detection of DNA copy number variation within regions of pronounced genomic complexity, which constitutes an improvement over available technologies. PMID- 16713173 TI - Pathogenetic transition in the morphology of abnormal sperm in the testes and the caput, corpus, and cauda epididymides of male rats after treatment with 4,6 dinitro-o-cresol. AB - In order to elucidate the pathogenesis of tailless sperm, 4,6-dinitro-o-cresol (DNOC) was administered to Jcl:SD male rats at daily oral doses of 0, 10 or 15mg/kg for 5 days. Sperm were collected from the caput, corpus, and cauda epididymides on days 1, 7 and 14 after the last dosing (D1, D7 and D14, respectively), counted and examined morphologically by phase-contrast and scanning electron microscopy. The incidence of abnormal sperm was significantly increased in the DNOC 15mg/kg group. On D1, peeled sperm (loss of mitochondrial sheath at the proximal end of the middle piece) was frequently observed in the caput epididymides, whereas sperm in the corpus and cauda epididymides had normal morphology. Distribution of the peeled sperm changed as time passed and the corpus epididymides showed a peak incidence on D7. On D14, the highest incidence of abnormal sperm was observed in the cauda epididymides, where the major abnormality was tailless. Similar effects were also found in the 10mg/kg group but were less potent. Transmission electron microscopy of testicular sperm on D1 revealed the presence of elongated spermatids that lacked the mitochondrial sheath at the proximal end of the middle piece, although the round and elongating spermatids looked normal. These results suggest that DNOC exposure of male rats primarily causes partial loss of the mitochondrial sheath in the testicular elongated spermatids, and that the affected sperm become tailless by D14 after reaching the cauda epididymides. PMID- 16713172 TI - Comparison of linkage disequilibrium patterns between the HapMap CEPH samples and a family-based cohort of Northern European descent. AB - The International HapMap Consortium has determined the linkage disequilibrium (LD) patterns of four major human populations. The aim of our investigation was to compare the LD patterns of the HapMap CEPH (Centre d'Etude du Polymorphisme Humain) samples with a family-based cohort of similar ancestry to determine its usefulness as a reference population for disease association studies. We examined four genomic regions on chromosomes 7q, 12p, and 14q totaling 14.3 Mb, initially identified in our linkage study of obesity and the metabolic syndrome. Near identical patterns of LD were detected in both populations. Furthermore, tagSNPs selected based on the HapMap CEPH cohort data capture over 98% of the variants at an r2 > 0.8 in the disease cohort. This confirms the usefulness of the CEPH cohort of the HapMap as a reference sample for further investigations into the genomic variation of populations of Northern European descent. PMID- 16713174 TI - Low doses of dietary lead are associated with a profound reduction in the time to the onset of puberty in female mice. AB - We report results from a replication in second and third generation female mice of accelerated time to puberty associated with low Pb exposure levels . Mice in the 2nd generation study are offspring of mice from the initial study; the 3rd generation mice are offspring from mice in the 2nd generation study. For each generation the time to puberty onset was markedly influenced by exposure to dietary lead. Modest increases in blood lead concentration from a normal background of 2-3 to 8-13 micro g/dl delayed the onset of puberty by 10-20% from a normal of 33-35 days to about 40-43 days; reducing blood lead from 2-3 to 0.7 micro g/dl was associated with profound acceleration of puberty to 21 days, an enhancement by over 30%. This dose-response relationship, which replicates previous novel findings, has possible ecological as well as public health significance and indicates that lead is able to induce biologically significant changes at blood lead levels previously thought to be without effect. PMID- 16713175 TI - Effects of in utero and lactational exposure to 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD) on rat follicular steroidogenesis. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a widespread environmental pollutant and causes adverse effects on female reproduction when administered to rats. Our aims were to study effects of gestational and lactational exposure to TCDD on ovarian steroidogenesis and steroidogenic enzyme expression of offspring on postnatal day (PND) 14 in the rat and sensitivity of enzymatically isolated ovarian follicles to TCDD in vitro. Synthetic estrogen diethylstilbestrol (DES) was used as a treatment control. Serum progesterone (P4) level in offspring increased significantly on PND 14 in the TCDD (1 microg/kg)-exposed group while body weight, FSH and E2 levels were not changed. In ovarian follicles of offspring on PND 14 in the TCDD-exposed groups, protein expression of cytochrome P-450 aromatase, cytochrome P-450 cholesterol side-chain cleavage, steroidogenic acute regulatory protein, 3beta-hydroxy-steroid-dehydrogenase/Delta(5)-Delta(4) isomerase type 1, or P4 receptor was not affected. TCDD decreased E2 and P4 production in ex vivo follicle culture. DES at a dose level of 0.1mg/kg was dystocic while a dose 0.02 mg/kg increased ovarian ex vivo E2 and testosterone production without affecting P450arom activity indicating stimulation of early steps of steroidogenic pathway. Data suggests that TCDD has multiple targets in ovarian steroidogenesis, but the inhibitory action represented as decreased follicular steroid hormone production ex vivo is not apparent at the ovarian protein expression. Furthermore, TCDD had no direct effect on immature rat ovarian steroidogenesis in vitro suggesting that the follicle culture method is not a sensitive method to study the mechanisms of TCDD action. PMID- 16713176 TI - Effect of two insecticides and two herbicides on the porcine sperm motility patterns using computer-assisted semen analysis (CASA) in vitro. AB - The recent decline in sperm concentration observed in men has developed over a short period of time, suggesting that it could be the result of environmental factors. The present study has evaluated the effects of insecticides Malathion and Diazinon, and herbicides Atrazine and Fenoxaprop-Ethyl on porcine sperm viability and motility patterns in vitro using the eosin-nigrosin staining and a computer-assisted semen analyzer (CASA), respectively. Malathion and Fenoxaprop Ethyl exerted more deleterious effects than Diazinon and Atrazine. Progressive sperm motility was strongly affected whereas the effect on sperm viability was less pronounced. This suggests that a reduction of sperm motility is not necessarily the result of sperm death. Since sperm motility is dependent on energy metabolism the mechanism of action of these pesticides might be mediated at the level of the mitochondrion, producing a delay in motility and eventual cell death. PMID- 16713177 TI - Differences in null points between the left and right ventricles in contrast enhanced inversion recovery MR imaging in patients with cardiac diseases. AB - Delayed contrast-enhanced inversion recovery (IR) gradient-echo MR imaging has been applied to several cardiac diseases, including myocarditis, sarcoidosis, hypertrophic cardiomyopathy, and myocardial damages induced by medical procedures. Although a preliminary study has indicated the usefulness of this imaging for the detection of right ventricular (RV) myocardial damage associated with arrhythmogenic right ventricular cardiomyopathy, the null points of the RV myocardium have not been assessed on contrast-enhanced IR MR imaging. In this study, the null points of the RV and left ventricular (LV) myocardia were evaluated using an IR fast multi-shot echo-planar imaging (Look-Locker sequence) in 26 patients with various cardiac diseases. In nine of the 26 patients, the null points of the RV myocardium were shorter than those of the LV myocardium in the Look-Locker sequence. The RV myocardial signals were significantly higher than the LV myocardial signals in delayed contrast-enhanced MR images. Thus, more attention should be paid to evaluation of the late enhancement of the RV myocardium, and delayed contrast-enhanced MR imaging with a shorter inversion time may be required in some cases. PMID- 16713178 TI - HLA-associated production of anti-DFS70/LEDGF autoantibodies and systemic autoimmune disease. AB - Autoantibodies against DFS70/LEDGF, which is also known as an important partner of HIV-1 integrase, are found in 10% of healthy Japanese people, but in only approximately 2% of patients with systemic autoimmune disease (SAD). We wished to characterize the association of HLA class II alleles with the presence of autoantibodies against this molecule. MHC class II genes (DR, DQ, and DP alleles) were analyzed by the polymerase chain reaction-sequence specific primer method in 24 individuals with anti-DFS70 antibodies. The frequencies of HLA-DRB1*0410, DQB1*0402, and -DPB1*0301 were increased in anti-DFS70 Ab-positive patients, while HLA-DQB1*0302 was decreased compared to Japanese controls. All anti-DFS70 Ab-positive individuals expressed at least one HLA-DQB1 allele with an aspartic acid at residue 57. The immunogenetic background of Japanese individuals with anti-DFS70 antibodies differs from that of patients with SAD. HLA class II genes influence the production of anti-DFS70 antibodies among individuals with various clinical manifestations. PMID- 16713179 TI - Short administration of polyclonal anti-T cell antibody (ALS) in NOD mice with extensive insulitis prevents subsequent development of autoimmune diabetes. AB - Treatment of overtly diabetic NOD mice with antilymphocyte serum (ALS), a polyclonal anti-T cell antibody, leads to cure of diabetes. Here, we investigated whether ALS-treatment of NOD mice after development of extensive insulitis prevents onset of diabetes. Female NOD mice were treated with two doses of ALS at 14, 19 or 23 weeks of age. No further treatment was given. In untreated female NOD mice, diabetes developed starting at 13 weeks and reached 68% by 37 weeks. ALS-treatment at 14, 19 or 23 weeks when histology showed progressive insulitis completely prevented onset of overt diabetes in 9/12, 11/12 or 12/12 mice, respectively. Intraperitoneal glucose tolerance tests in 43 week-old ALS-treated, diabetes-free mice showed a normal pattern. Co-adoptive transfer of lymphoid cells prepared from ALS-treated diabetes-free mice together with splenocytes from overtly diabetic NOD mice resulted in marked delay in diabetes onset in NOD.SCID mice, suggesting the presence of autoimmune regulatory cells in ALS-treated mice. Autoimmune regulatory cells were CD4(+)CD25(+), but not CD4(+)CD25(-), T cells. Thus, treatment of euglycemic individuals who already show signs of autoimmune diabetes with a short course of polyclonal anti-T cell antibody may effectively prevent onset of type 1 diabetes mellitus. PMID- 16713180 TI - N-3 fatty acids modulate Th1 and Th2 dichotomy in diabetic pregnancy and macrosomia. AB - We assessed the implication of Th (helper)-cells and the modulation of the Th1/Th2 dichotomy by n-3 polyunsaturated fatty acids (PUFA) in type I diabetic pregnancy (DP) and macrosomia. Female gestant rats fed a standard diet or n-3 PUFA regimen were rendered diabetic by administration of five low doses of streptozotocin. The macrosomic (MAC) offspring were sacrificed at the age of 90 days. The mRNAs of IL-2 and IFN-gamma (Th1 cytokines) and IL-4 (Th2 cytokine) were downregulated in the pancreas and spleen of diabetic pregnant rats. The levels of IL-10 mRNA, another Th2 cytokine, were unchanged in the spleen or upregulated in the pancreas of these animals. Feeding an n-3 PUFA diet to rats with DP upregulated IL-10 mRNA in the pancreas and IL-4 and IL-10 mRNA in the spleen. In MAC offspring, high expression of IL-2 and IFN-gamma mRNA, but not of Th2 cytokines, was observed. The n-3 PUFA diet diminished Th1 mRNA quantities and increased the levels of IL-4, but not of IL-10, mRNA in MAC offspring. Our study shows that DP is associated with a decreased Th1 phenotype and IL-4 mRNA expression in the pancreas and spleen, and an n-3 PUFA diet upregulates Th2 profile. In MAC offspring, the Th1 phenotype is upregulated and an n-3 PUFA diet downregulates this phenomenon. PMID- 16713181 TI - Molecular mechanism of cofilin dephosphorylation by ouabain. AB - We previously reported that phosphorylated cofilin-triosephosphate isomerase (TPI) complex interacts with Na,K-ATPase and enhances the pump activity through the phosphorylation of cofilin via Rho-mediated signaling pathway. In this study, we tested the hypothesis that the dephosphorylation of cofilin may be induced through Na,K-ATPase inhibition by ouabain. The phosphorylation level of cofilin by ouabain which decreases in a time- and dose-dependent manner in various human cell lines, remains unchanged by pretreatment with Src inhibitor, PP2; epidermal growth factor receptor (EGFR) inhibitor, AG1478; Raf-1 kinase (Raf) inhibitor, GW5074; and ERK kinase (MEK) inhibitor, PD98059, and by transfection of Ras dominant negative mutant (RasN17). This suggests that ouabain dephosphorylates cofilin through the Src/EGFR/Ras/Raf/MEK pathway. Ouabain activates Ras/Raf/MEK pathway, but down-regulates Rho kinase (ROCK)/LIM kinase (LIMK)/cofilin pathway, implying that there may be a cross-talk by ouabain between the Ras/Raf/MEK and the ROCK/LIMK/cofilin pathways. Immunofluorescence and flow cytometry suggest that ouabain-induced active form of cofilin may be involved in cytoskeletal reorganization and cell volume regulation. Thus, these findings demonstrate a new molecular mechanism for the dephosphorylation of cofilin through the inhibition of Na,K-ATPase by ouabain. PMID- 16713182 TI - Epidermal growth factor receptor and DNA double strand break repair: the cell's self-defence. AB - The purpose of this review is to discuss the relation between the repair of DNA double strand breaks (DSB)--the main lethal lesion inflicted by ionising radiation-and the function of receptors of epidermal growth factor (EGFR) and similar ligands (other members of the ERBB family). The reviewed experimental data support the assumption that in mammalian cells, one consequence of EGFR/ERBB activation by X-rays is its internalisation and nuclear translocation together with DNA-dependent protein kinase (DNA-PK) subunits present in lipid rafts or cytoplasm. The effect of EGFR/ERBB stimulation on DSB rejoining would be due to an increase in the nuclear content of DNA-PK subunits and hence, in activity increase of the DNA-PK dependent non-homologous end-joining (D-NHEJ) system. Such mechanism explains the radiosensitising action of "membrane-active drugs", hypertonic media, and other agents that affect nuclear translocation of proteins. Also, one radiosensitising effect of the recently introduced into clinical practice EGFR/ERBB inhibitors would consist on counteracting the nuclear translocation of DNA-PK subunits. In result, D-NHEJ may be less active in inhibitor-treated cells and this will contribute to an enhanced lethal effect of irradiation. The reviewed observations point to a heretofore not understood mechanism of the cell's self-defence against X-rays which can be exploited in combined radio- and chemotherapy. PMID- 16713183 TI - Surgery for epilepsy: assessing evidence from observational studies. AB - The effectiveness of epilepsy surgery is mostly based on the results of observational studies because an experimental design in surgical patients has several ethical implications. However, observational studies suffer from methodological drawbacks, which prevent meaningful conclusions and represent a serious limitation when data from different reports are pooled for systematic reviews and meta-analyses. These include the retrospective design, the use of referral populations, the small sample size, the enrolment of patients at differing inception points, the use of differing inclusion criteria, differing definitions of prognostic predictors and outcome measures, the unmasked assessment of outcome, the short follow-up, the changing technologies and surgical procedures, and the inadequate statistical methods. With these limitations in mind, a systematic review was performed of epilepsy surgery studies aiming at identifying the positive and negative prognostic predictors of surgical outcome. In this review, extent of surgical resection, abnormal MRI, mesial temporal sclerosis, febrile seizures, EEG/MRI concordance, and tumor were in decreasing order the principal indicators of the success of surgery of epilepsy. By contrast, intracranial monitoring and post-operative discharges tended to predict an unfavorable prognosis. Although the heterogeneity of the study results was fairly low, the results may be affected by pooling of data from heterogeneous reports (different patient series assessed with differing methods) or unexamined or unknown confounders. These data must be considered preliminary and cannot replace well-conducted prognostic studies (representative study populations, well-defined inception cohorts, satisfactory and complete follow-up, prospective design, and standard definition of prognostic factors). PMID- 16713184 TI - The speed of visual attention in schizophrenia: electrophysiological and behavioral evidence. AB - Schizophrenia is characterized by a substantial slowing of manual response times and by impairments in attention. However, prior research has not investigated whether attention itself is slowed in schizophrenia, and this was the goal of the present study. In Experiment 1, the N2pc component of the event-related potential waveform-an electrophysiological correlate of the focusing of attention-was recorded from 24 schizophrenia spectrum patients and 13 control subjects. Although behavioral response times were delayed by over 100 ms in the patient group, the onset latency of the N2pc component was virtually identical across groups, and no reduction in N2pc amplitude was observed in the patient group. In Experiment 2, a new cueing paradigm was developed to provide a behavioral measure of the speed of attention in 22 schizophrenia spectrum patients and 13 control subjects. We found that the average time required to allocate attention to a cued location was only 19 ms greater for the patient group than for the control group, with most patients within the range of the control subjects. Together, these experiments revealed little or no slowing of the allocation of visual-spatial attention in patients with schizophrenia. Thus, the mechanisms responsible for allocating attention to salient visual targets appear to be largely unaffected by the illness, and the well documented slowing of manual response times in schizophrenia cannot easily be explained by a slowing of attention. PMID- 16713185 TI - D2/D3 dopamine receptor binding with [F-18]fallypride in thalamus and cortex of patients with schizophrenia. AB - BACKGROUND: Abnormalities in the dopaminergic system are implicated in schizophrenia. [F-18]fallypride is a highly selective, high affinity PET ligand well suited for measuring D2/D3 receptor availability in the extrastriatal regions of the brain including thalamus, prefrontal, cingulate, and temporal cortex, brain regions implicated in schizophrenia with other imaging modalities. METHODS: Resting [F-18]fallypride PET studies were acquired together with anatomical MRI for accurate coregistration and image analysis on 15 drug naive schizophrenics (10 men, 5 women, mean age 28.5 years) and 15 matched controls (9 men, 6 women, mean age 27.4 years). Dopamine D2/D3 receptor levels were measured as binding potential (BP). The fallypride BP images of each subject were spatially normalized and subsequently smoothed for group comparison. Measures of significance between the schizophrenic and control groups were determined using statistical parametric mapping (SPM). The medial dorsal nucleus and pulvinar were also traced on coregistered MRI for detailed assessment of BP in these regions. RESULTS: The thalamus of patients with schizophrenia had lower [F-18]fallypride BP than normal controls and this was the brain area with the greatest difference (range -8.5% to -27.2%). Left medial dorsal nucleus and left pulvinar showed the greatest decreases (-21.6% and -27.2% respectively). The patients with schizophrenia also demonstrated D2/D3 BP reduction in the amygdala region, cingulate gyrus, and the temporal cortices. CONCLUSIONS: These findings suggest that drug naive patients with schizophrenia have significant reductions in extrastratial D2/D3 receptor availability. The reductions were most prominent in regions of the thalamus, replicating other studies both with high affinity D2/D3 ligands and consistent with FDG-PET studies, further supporting the hypothesis of thalamic abnormalities in this patient population. PMID- 16713186 TI - Changing the outcome of toxoplasmosis in bone marrow transplant recipients. PMID- 16713187 TI - First description of CTX-M beta-lactamase-producing clinical Escherichia coli isolates from Egypt. AB - We studied the presence of beta-lactamases with an extended spectrum of activity in clinical Escherichia coli isolates from Cairo, Egypt. Forty-six E. coli isolates were collected from patients with urinary tract infections at a university hospital in 2001. Phenotypic characterisation identified a very high extended-spectrum beta-lactamase (ESBL) rate of 60.9%. Pulsed-field gel electrophoresis and plasmid profiles revealed eight different clonal groups. All ESBL producers were polymerase chain reaction-positive for blaTEM and blaCTX-M genes. Within the CTX-M family, three different enzymes, CTX-M-14, CTX-M-15 and CTX-M-27, were found. The ESBL producers carried multiple plasmids and further plasmid-encoded resistances. In several strains, genes for up to six aminoglycoside-modifying enzymes were detected. A linkage to fluoroquinolone resistance was not observed. This study confirms the high rate of ESBLs in Egypt and further demonstrates the worldwide spread of genes coding for CTX-M enzymes in clinical isolates. PMID- 16713188 TI - Cytoskeleton reassembly in cardiomyocytes infected by Trypanosoma cruzi is triggered by treatment with ergosterol biosynthesis inhibitors. AB - We have previously shown that Trypanosoma cruzi-infected cardiomyocytes present alterations in cytoskeletal organisation in vitro. The remarkable change in the host cell cytoskeleton opened up the question of whether treatment of infected cells with antitrypanosomal compounds, such as ergosterol biosynthesis inhibitors (EBIs), allows the reconstruction of myofibrils and the microtubule network, restoring the cell biological function. Therefore, 48-h-old T. cruzi-infected cardiomyocyte cultures were treated with 10 nM ketoconazole or posaconazole and cytoskeletal remodelling of the host cells was analysed by indirect immunofluorescence assay. Both compounds displayed a potent antiparasitic effect and dramatically reduced the infection ratio. After 120 h of treatment, actin polygonal configuration was frequently visualised in the host cell cytoplasm, suggesting the initial stage of actin framework restoration. Rearrangement of myofibrils and the microtubule network was achieved 168 h after the start of drug treatment. Our data demonstrate that the trypanocidal effect of EBIs lead to reconstruction of the cytoskeleton of T. cruzi-infected cardiomyocytes in vitro. PMID- 16713189 TI - Brevinin-1BYa: a naturally occurring peptide from frog skin with broad-spectrum antibacterial and antifungal properties. AB - Brevinin-1BYa (FLPILASLAAKFGPKLFCLVTKKC) is a cationic alpha-helical peptide containing an intramolecular disulphide bridge that is present in skin secretions of the foothill yellow-legged frog Rana boylii. A synthetic replicate of the peptide showed growth inhibitory activity against a range of reference strains of Gram-positive and Gram-negative bacteria, against clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) (minimum inhibitory concentration (MIC)=2.5 microM), and against reference strains and clinical isolates of the opportunistic yeast pathogens Candida albicans, Candida tropicalis, Candida krusei and Candida parapsilosis (MICor=32 microg/mL), sulphamethoxazole/trimethoprim (SMX/TMP) (resistant>or=4 microg/mL), nitrofurantoin (resistant>or=128 microg/mL), ciprofloxacin (resistant>or=4 microg/mL) and levofloxacin (resistant>or=8 microg/mL) resistance breakpoints used were those published by the CLSI. Of the 1142 E. coli collected, 75.5% (862) were collected from the USA and 280 (24.5%) were from Canada. Patient demographics revealed a mean age of 48.1 years (range, 2 months to 99 years), with female patients representing 79.4% of patients and males representing 20.6%. Overall, resistance to ampicillin was 37.7%, followed by SMX/TMP (21.3%), nitrofurantoin (1.1%), ciprofloxacin (5.5%) and levofloxacin (5.1%). Resistance rates for all antimicrobials were higher in US medical centres compared with Canadian centres (P<0.05). Fluoroquinolone resistance was highest in patients>or=65 years of age (P<0.05). Resistance rates demonstrated considerable geographic variability both in the USA and Canada. This study reports higher rates of antibiotic resistance in US versus Canadian outpatient urinary isolates of E. coli and demonstrates the continuing evolution of resistance to antimicrobial agents. PMID- 16713190 TI - Antibacterial activities of inorganic agents on six bacteria associated with oral infections by two susceptibility tests. AB - The antibacterial effects of six inorganic antibacterial agents were assessed using broth dilution and agar dilution tests on six pathogenic bacteria associated with oral infectious diseases: Streptococcus mutans (ATCC 25175), S. mutans (Ingbritt), Actinomyces viscosus (ATCC 15987), Lactobacillus casei (ATCC 393), Staphylococcus aureus (ATCC 29213) and Candida albicans (ATCC 90028). The results of the broth dilution test were significantly lower than those of the agar dilution test (F=38.290; P<0.01). The six inorganic agents notably inhibited the growth of tested common oral bacteria in vitro. Among them, Longbei inorganic antibiotic powder was the strongest antibacterial agent, followed by ZnO whisker antibacterial complex (ZnOw) AT-83, IONPURE-H, basic magnesium hypochlorite, ZnOw AT-88 and Antim-AMS2. The broth dilution test appears to be more suitable for testing insoluble inorganic agents. PMID- 16713192 TI - Potential role of aerosolized amphotericin B formulations in the prevention and adjunctive treatment of invasive fungal infections. AB - The incidence of invasive fungal infections (IFIs) continues to increase, largely due to the steady rise in the number of at-risk patients and the increased use of aggressive immunosuppressant agents. Many available treatments are often limited by concerns about efficacy, safety, drug interactions, and/or cost. Owing to the poor treatment outcomes of immunosuppressed patients with IFIs, new preventative and treatment strategies are being investigated. Among these are the aerosolized formulations of amphotericin B. Published experience with the use of aerosolized amphotericin B deoxycholate (AmBd) in the prevention of IFIs has raised concerns regarding challenges in drug administration and tolerability. However, evolving data regarding administration of lipid-based formulations of amphotericin B indicate potential advantages over AmBd in the prevention and adjunctive treatment of IFIs. PMID- 16713193 TI - Antifungal combination treatment: a future perspective. AB - The use of antifungal combination therapy is a new clinical approach for combating fungal infections. Although few conclusive clinical studies have been performed to date, data exist that strongly suggest that combination therapy would benefit specific patient subgroups (e.g. stem-cell transplant recipients). This paper examines the evidence available from in vitro, experimental, and clinical studies, and seeks to discover viable clinical strategies for this promising new therapeutic approach. PMID- 16713194 TI - Serotonin transporter characteristics in lymphocytes and platelets of male aggressive schizophrenia patients compared to non-aggressive schizophrenia patients. AB - A large body of literature indicates that disturbances of central serotonin (5 HT) function play an important role in aggressive behavior. Results from open label and placebo-controlled trials as well as the reported inverse relationship between 5-HT function and aggression in human subjects, suggest that reduced 5-HT activity is associated with aggressive behavior. The activity of the 5-HT transporter (5-HTT), as determined by [(3)H]5-HT uptake to blood lymphocytes, was measured in 20 currently aggressive and 20 non-aggressive male schizophrenia patients. In addition, the pharmacodynamic characteristics of platelet 5-HTT were assessed by [(3)H]citalopram binding. There were no significant differences in the density (B(max)) of platelet [(3)H]citalopram binding sites between the two groups. Similarly, the dissociation constant (K(d)) values were indistinguishable. The maximum uptake velocity (V(max)) of [(3)H]5-HT to fresh lymphocytes and the K(m) values of the 5-HT to the transporter were significantly higher in currently aggressive compared to the non-aggressive schizophrenia patients. The association of high V(max) values with current aggressive behavior provides further support to the involvement of the 5-HTT in aggressive behavior as well as to the efficacy of 5-HTT blockers in the control of aggression. The role of the various components of the serotonergic system in the pathophysiology and treatment of aggressive behavior in schizophrenia needs to be further evaluated. PMID- 16713195 TI - Oxidative stress in ALS: a mechanism of neurodegeneration and a therapeutic target. AB - The cause(s) of amyotrophic lateral sclerosis (ALS) is not fully understood in the vast majority of cases and the mechanisms involved in motor neuron degeneration are multi-factorial and complex. There is substantial evidence to support the hypothesis that oxidative stress is one mechanism by which motor neuron death occurs. This theory becomes more persuasive with the discovery that mutation of the anti-oxidant enzyme, superoxide dismutase 1 (SOD1), causes disease in a significant minority of cases. However, the precise mechanism(s) by which mutant SOD1 leads to motor neuron degeneration have not been defined with certainty, and trials of anti-oxidant therapies have been disappointing. Here, we review the evidence implicating oxidative stress in ALS pathogenesis, discuss how oxidative stress may affect and be affected by other proposed mechanisms of neurodegeneration, and review the trials of various anti-oxidants as potential therapies for ALS. PMID- 16713196 TI - Translating preclinical insights into effective human trials in ALS. AB - Amyotrophic lateral sclerosis (ALS) is a rapidly progressive, adult-onset neurodegenerative disease characterized by selective dysfunction and death of motor neurons in the brain and spinal cord. The disease is typically fatal within 3-5 years of symptom onset. There is no known cure and only riluzole, which was approved by the FDA in 1996 for treatment of ALS, has shown some efficacy in humans. Preclinical insights from model systems continue to furnish ample therapeutic targets, however, translation into effective therapies for humans remains challenging. We present an overview of clinical trial methodology for ALS, including a summary rationale for target selection and challenges to ALS clinical research. PMID- 16713197 TI - The ADAM metalloprotease Kuzbanian is crucial for proper heart formation in Drosophila melanogaster. AB - We have screened a collection of EMS mutagenized fly lines in order to identify genes involved in cardiogenesis. In the present work, we have studied a group of alleles exhibiting a hypertrophic heart. Our analysis revealed that the ADAM protein (A Disintegrin And Metalloprotease) Kuzbanian, which is the functional homologue of the vertebrate ADAM10, is crucial for proper heart formation. ADAMs are a family of transmembrane proteins that play a critical role during the proteolytic conversion (shedding) of membrane bound proteins to soluble forms. Enzymes harboring a sheddase function recently became candidates for causing several congenital diseases, like distinct forms of the Alzheimer disease. ADAMs play also a pivotal role during heart formation and vascularisation in vertebrates, therefore mutations in ADAM genes potentially could cause congenital heart defects in humans. In Drosophila, the zygotic loss of an active form of the Kuzbanian protein results in a dramatic excess of cardiomyocytes, accompanied by a loss of pericardial cells. Our data presented herein suggest that Kuzbanian acts during lateral inhibition within the cardiac primordium. Furthermore we discuss a second function of Kuzbanian in heart cell morphogenesis. PMID- 16713198 TI - Quantitative analysis of 17O exchange and T1 relaxation data: application to zirconium tungstate. AB - The theoretical basis behind a recent quantitative analysis of 17O exchange in ZrW2O8 [M.R. Hampson, J.S.O. Evans, P. Hodgkinson, J. Am. Chem. Soc. 127 (2005) 15175-15181] is set out. Despite the complexities of combining the multi exponential relaxation of half-integer quadrupolar nuclei with chemical exchange, it is shown how magnetisation transfer experiments can be analysed to obtain estimates of absolute exchange rates. The multi-exponential relaxation is best modelled using a magnetic mechanism, i.e. the rapid T1 relaxation observed, particularly at high temperatures, can be directly related to the relatively high degree of 17O labelling employed. The combination of the 1D EXSY results with T1 values as a function of temperature provides exchange rates and activation barriers over a wide temperature range (40-226 degrees C). PMID- 16713199 TI - A comparison of morphology and surface energy characteristics of sulfathiazole polymorphs based upon single particle studies. AB - The morphological, adhesion and surface energetic properties of three sulfathiazole polymorphs (III, IV and polymorph I prepared from both acetone and methanol, designated I-ace and I-met, respectively) produced using Nektar supercritical fluid (SCF) technology have been characterized using scanning electron microscopy (SEM) and atomic force microscopy (AFM). Surface roughness values for each polymorph were determined at different length scales. At sample sizes less than 1micromx1microm the polymorphs rank in terms of roughness as follows: I-met>I-ace approximately equal to IV>III. At the larger scales the polymorphs rank in terms of roughness as follows: I-met>III>I-ace approximately equal to IV. The surface energies for polymorphs determined against graphite (HOPG) and particles of the same polymorph were, respectively, I-met: 0.99mJm(-2) (S.D. 1.25mJm(-2)), 3.09mJm(-2) (S.D. 2.67mJm(-2)); I-ace: 309mJm(-2) (S.D. 329mJm(-2)), 16mJm(-2) (S.D. 11mJm(-2)); III: 1.17mJm(-2) (S.D. 1.5mJm(-2)), 5.4mJm(-2) (S.D. 3.6mJm(-2)); IV: 20.35mJm(-2) (S.D. 28.5mJm(-2)), 16.8mJm(-2) (S.D. 9.6mJm(-2)). In terms of surface energies the polymorphs hence rank I ace>IV>III approximately equal to I-met (HOPG adhesion measurements) and IV approximately equal to I-ace>III>I-met (particle cohesion measurements). Consideration of contacting asperities and surface roughness was shown to have limited effect on the surface energies, and instead the differences were ascribed to variations in the surface chemistry as a result of changes in crystallization mechanisms. PMID- 16713200 TI - Particle swarm algorithm trained neural network for QSAR studies of inhibitors of platelet-derived growth factor receptor phosphorylation. AB - The multilayer feed-forward artificial neural network (ANN) has been widely used in QSAR studying. Back-propagation algorithm (BP) and the use of evolutionary search as an ANN training method has some limitations associated with overfitting, local optimum problems and slow convergence rate. In this paper, particle swarm optimization (PSO) as a relatively new optimization technique has been used in ANN training. Compared to ANN trained by BP algorithm and evolutionary search, ANN training by PSO algorithm (PSONN) show satisfactory performance, converges quickly towards the optimal position and can avoid overfitting in some extent. The PSONN has been testified by using in QSAR modeling for inhibitory activity of 4-[4-(N-substituted (thio) carbamoyl)-1 piperazinyl]-6,7-dimethoxyquinazoline derivatives. PMID- 16713202 TI - Synthesis of radiolabelled 1-trichloromethyl-1,2,3,4-tetrahydro-beta-carboline (TaClo), a neurotoxic chloral-derived mammalian alkaloid, and its biodistribution in rats. AB - Chloral hydrate, the oldest synthetic hypnotic, is still among the most common agents used for conscious sedation of infants and children. Chloral (Clo) spontaneously condenses with the biogenic amine tryptamine (Ta) giving rise to the endogenous formation of 1-trichloromethyl-1,2,3,4-tetrahydro-beta-carboline (TaClo). TaClo constitutes a mammalian alkaloid and a potent neurotoxin, which is capable of inducing a slowly developing degeneration of the dopaminergic system in rats. Due to the late onset of parkinsonian-type symptoms after TaClo administration, this agent has been postulated to be a potential natural inducer of Parkinson's disease. In order to elucidate its pharmacokinetics, its tissue distribution and excretion profile, radiolabelled [3-14C]-TaClo was prepared in a convenient four-step synthetic pathway. Studies on rats intraperitoneally treated with a single dose of 2.1 mg/kg (0.6 microCi/kg) of [3-14C]-TaClo.HCl (specific activity: 0.28 microCi/mg) revealed the radioactivity to be rapidly incorporated with the highest concentrations of 14C found in the excretory organs. [3-14C] TaClo was poorly absorbed systemically, as indicated by the very low plasma radioactivity levels. Maximum levels of 14C were reached between 2 and 6 h postdose, with the exception of large intestine peaking at 12 h after dosing. Total mean percent recovery of the radioactive dose was about 96% within the 48-h period examined. Urinary excretion accounted for ca. 34.5%, with the majority of the applied dose being eliminated by the hepatobiliary pathway. PMID- 16713201 TI - Permeability and swelling studies on free films containing inulin in combination with different polymethacrylates aimed for colonic drug delivery. AB - The aim of this study was to assess some permeability and swelling characteristics of free films prepared by combination of inulin as a bacterially degradable system and time- or pH-dependent polymers as a coating formulation for colonic drug delivery. Different free films were prepared by casting and solvent evaporation method. Formulations containing inulin with Eudragit RS, Eudragit RL, Eudragit RS-Eudragit RL, Eudragit FS and Eudragit RS-Eudragit S with different ratios of inulin were prepared. After preparation, free films were evaluated by water vapor transmission test, swelling experiment and permeability to indomethacin and theophylline in different media. Formulations containing Eudragit FS had high resistance to water vapor permeation; but were unable to protect premature swelling and drug release in simulated small intestine media. Also, combination of Eudragit RS and Eudragit S had no suitable characteristics for colon delivery. However, Eudragit RS and Eudragit RL in combination with inulin made free films which had more swelling and permeation of drug in the colonic medium rather than the other media. It was shown that formulations containing sustained release polymethacrylates in combination with inulin have more potential as a coating system for specific colon delivery compared with pH dependent polymers. PMID- 16713203 TI - Brain imaging tools in neurosciences. AB - In this chapter brain imaging tools in neurosciences are presented. These include a brief overview on single-photon emission tomography (SPET) and a detailed focus on positron emission tomography (PET) and functional magnetic resonance imaging (fMRI). In addition, a critical discussion on the advantages and disadvantages of the three diagnostic systems is added. Furthermore, this article describes the image analysis tools from visual analysis over region-of-interest technique up to statistical parametric mapping, co-registration methods, and network analysis. It also compares the newly developed combined PET/CT scanner approach with established image fusion software approaches. There is rapid change: Better scanner qualities, new software packages and scanner concepts are on the road paved for an amply bright future in neurosciences. PMID- 16713204 TI - Therapy-induced plasticity of cognitive functions in MS patients: insights from fMRI. AB - Multiple sclerosis (MS) is an inflammatory disease of the central nervous system whose pathological mechanisms are still not completely understood. Physical as well as cognitive deterioration are consequences within the disease process that have an extensive impact on the patient's quality of life. Therefore, understanding the functional background of spontaneous as well as induced remission is of high relevance. Studies on visualization of therapeutic effects of pharmacological or cognitive treatment by functional magnetic resonance imaging (fMRI) are still rare. From fMRI studies on focal brain lesions hypotheses on mechanisms of brain reorganization can be derived. This contribution will first give an overview of the existing studies using fMRI in MS, on cognitive decline, on cognitive treatment studies and its therapeutic effects on behavioural readouts in MS, and on therapy-induced brain plasticity and its possible visualization by fMRI. Results of a study on correlating the effects of cognitive training with changes in brain organization in patients with mild to severe cognitive impairment will be reported. PMID- 16713205 TI - [HIV transmission from surrogate breast feeding: 2 case reports from Bangui]. PMID- 16713206 TI - [When parents require or refuse neonatal intensive care: one French neonatologist's opinion]. PMID- 16713207 TI - [Immunoglobulin replacement therapy contribution in agammaglobulinemia: 10 case reports]. AB - Intravenous immunoglobulin (Ig IV) has been used for many years in the treatment of primary antibody deficiencies. We performed a retrospective study of the clinical features and outcome of agammaglobulinemia children who received prolonged Ig IV infusions. PATIENTS AND METHODS: Ten children, 9 male et 1 female, with agammaglobulinemia diagnosis were studied for the clinical manifestations before and during the Ig IV replacement therapy. Serum Ig levels were quantified by nephelometry. Circulating B ant T cells were counted by immunofluorescence labeling by monoclonal antibodies. T-cell functions were assessed by using mitogen and antigen -induced T-cell proliferation assays in vitro. Patients clinical status was evaluated respectively, before initiation and at every moment (when patients had an infection) of the replacement therapy. RESULTS: Ig IV therapy was performed for 866 cumulated months, median 108 months. The median Ig IV doses administered to the 10 patients was 500 mg/kg/month. Residual serum IgG mean level was 3,9 g/L. All patients had 99 bacterial infections/year before Ig IV, mainly respiratory tract infections (48,5%), and 4 patients had bronchiectasis before Ig replacement therapy. The number of infection/year fall to 25 during IgIV replacement, and the infection/patient/year rate decreases significantly. One patient developed an Echovirus 27 meningoencephalitis during this treatment. CONCLUSION: Ig IV therapy with residual IgG mean level of 3,9 g/l reduced significantly the rate of bacterial infections. The use of specific antibiotherapy and respiratory kinesitherapy led to a lower rate of respiratory tract infections, and the stabilisation of the bronchiectasis. However this intravenous replacement therapy does not protect against viral meningoencephalitis. PMID- 16713209 TI - [European network for investigation of medicinal products in children (Medichildren)]. PMID- 16713208 TI - [Evaluation of the treatments for the cerebral palsy child]. PMID- 16713210 TI - [Congenital esophageal stenosis due to tracheobronchial remnants: report of 2 cases and literature review]. AB - Congenital esophageal stenosis due to tracheobronchial remnants is a rare malformation whose diagnosis may be difficult. It is characterised by the abnormal presence of congenital tissue of tracheal origin in the esophageal wall, which is responsible for the narrowing of the esophagus. We report 2 cases whose treatment was surgical after failure of esophageal dilations. The presence of tracheal-bronchial tissue was confirmed by histological examination of the operative piece. Outcome was favourable and the final result was excellent. Recently, endoscopic ultrasonography has been proved useful in the diagnosis of congenital esophageal stenosis due to tracheobronchial remnants by showing the presence of cartilage, which explains the failure of dilation. The high rate of perforation in these cases is due to brutal fragmentation of the cartilaginous rings. Surgical resection of esophageal stenosis with the tracheobronchial tissue appears the only treatment susceptible to completely suppress the stenosis and its consequences. PMID- 16713211 TI - Lymphomas and leukemias in mice. AB - Lymphomas are among the most common tumors in many strains and stocks of mice, especially those used in safety assessment. CD-1, C57BL/6, B6C3F1 and B6;129 mice develop 10-50% incidences of lymphomas in aging mice. Most of the tumors are B cell lymphomas of the follicular type, arising in spleen, mesenteric lymph node and/or Peyer's patches. Lymphomas and leukemias may be induced by chemicals, retroviruses and irradiation. Genetics also play a major role in mouse lymphomagenesis and leukemogenesis. The most potent chemical carcinogens require only a single injection in young mice to induce a high incidence of lymphomas, often thymic T-cell lymphoblastic lymphomas. Several genetically engineered mouse lines have high incidences of these tumors. In 2-year carcinogenesis bioassays, increases of incidences of B-cell lymphomas (which occur naturally in controls) are indicative of less potent carcinogens. Classifications of the lymphomas and leukemias have evolved over the years. The practical WHO toxicologic pathology lymphoma and leukemia classification was developed by collaboration between the US STP, RITA, BSTP and JSTP. A more recent mouse lymphoma and leukemia classification was published by a committee of the US NCI Mouse Models of Human Cancers Consortium. This classification follows closely the more detailed human WHO classification and can be used for mouse models of lymphoma and leukemia. PMID- 16713213 TI - Preclinical safety assessment of a DNA vaccine using particle-mediated epidermal delivery in domestic pig, minipig and mouse. AB - DNA vaccination involves the direct injection of genes coding for specific antigenic proteins. One technique known as particle-mediated epidermal delivery (PMED) is a practical approach for epidermal delivery and provides a strong immune response. An important aspect of the preclinical safety assessment of DNA vaccines is the selection of a pharmacologically relevant animal model for the assessment of antigen expression, optimization of delivery and formulation of the plasmid. This paper describes a comparative study of domestic pig, minipig and mouse in regard to local tolerance and antigen expression of HIV immunotherapeutic using PMED. Pig/minipig is considered a good model for the safety assessment of DNA vaccines due to the similarity to human skin. Local reactions were evaluated at 10 min, 4, 24 and 48 h. Histology of administration sites revealed epidermal necrosis with associated dermal inflammation at 10 min and 4h, and subsequent regeneration with repair at 24 and 48 h. The degree and extent of these changes varied according to species. Domestic pig and minipig showed superficial epidermal necrosis and complete repair, while the mouse showed full-thickness epidermal necrosis and partial repair. Expression of HIV antigen was confirmed using immunohistochemistry in all three species at 4, 24 and 48 h. The results showed that PMED is an effective system for DNA vaccine delivery as demonstrated by the antigen expression seen as early as 4 h. PMID- 16713212 TI - Benzene metabolites induce apoptosis in lymphocytes. AB - Benzene is an important environmental pollutant with important health implications. Exposure to this aromatic hydrocarbon is associated with hematotoxicity, and bone marrow carcinogenic effects. It has been shown that benzene induces oxidative stress, cell cycle alterations, and programmed cell death in cultured cells. Hepatic metabolism of benzene is thought to be a prerequisite for its bone marrow toxicity. Nevertheless, there are no reports on the cellular effects of reactive intermediates derived from hepatic metabolism of benzene. Thus, the goal of this project was to determine the cellular alterations of benzene metabolites produced by the cultured hepatic cell line HepG2. Supernatants collected from these cells were applied to a culture of freshly isolated lymphocytes. A higher decrease in cell viability was found in cells exposed to these supernatants than to unmetabolized benzene. This viability decrease was due to apoptosis, as determined by Terminal deoxynucleotidyl Transferase Biotin-dUTP Nick End Labeling (TUNEL) assay and internucleosomal fragmentation of DNA. When supernatants were analyzed by HPLC, we found that not all the hydrocarbon was biotransformed, since a 28 microM concentration (37%) remained. The only metabolite found in the culture medium was muconic acid. The present results show that muconic acid derived from benzene metabolism is able to cooperate with the pollutant for the induction of apoptosis in rat lymphocytes. PMID- 16713214 TI - Regulatory aspects of immunotoxicology. AB - Since years, differences among the regulatory requirements on preclinical immunotoxicity testing for pharmaceuticals in the EU, Japan and US indicated a need for an internationally accepted approach. Requests for immunotoxicity investigations are also addressed by guidelines in non-drug areas. While some contain more detailed information in their requirements, other regulations comprise only vague descriptions for consideration of (non-intended) effects on the immune effects. Since 2002, the International Conference on Harmonisation (ICH) of Technical Requirements for Registration of Pharmaceuticals for Human Use put effort in the development of a harmonised approach for testing of immunosuppression and immunoenhancement. Consensus on the ICH S8 guideline on immunotoxicity testing for pharmaceuticals was achieved which now can be implemented into national regulations. The new concept contains in-depth testing, e.g., by functional tests in a concern/weight of evidence approach if the standard toxicity studies or other causes of concern give evidence of an immunotoxic potential or when the target populations are specifically vulnerable. It is expected that the progress on immunotoxicity testing reached by the ICH process will also have an impact on other regulatory areas. Additionally, the regulatory differences in testing requirements on immunotoxicity in other pharmaceutical areas including biotechnology-derived drugs, medicinal products and vaccines and in non-drug areas consisting of chemicals, agrochemicals or food additives are briefly highlighted. PMID- 16713215 TI - Restricted feeding may induce serous fat atrophy in male Gottingen minipigs. AB - Serous atrophy of the fatty tissue of bone marrow is occasionally seen in minipigs. It is not associated with compound toxicity, as it has been observed in both dosing and healthy control groups in toxicity studies, but the etiology and pathogenesis are unknown. However, a nutritional factor is suspected, since minipigs are generally diet restricted. In order to investigate this, newly weaned minipigs were fed different amounts of feed for nine months, after which the bone marrow was evaluated. Serous atrophy was observed in diet-restricted male minipigs fed on a diet based on the nutrient requirements of production pigs, but not in males fed on a diet specially designed for minipigs, at similarly restricted levels. No serous atrophy was observed in females. It was concluded that nutrient requirements of production pigs are not necessarily suitable for minipigs, as a diet based on these nutrient requirements caused total depletion of perirenal fat depots, as well as serous atrophy of the fatty tissue of bone marrow in male Gottingen minipigs at a restricted feeding level. PMID- 16713216 TI - Protective effect of Dunaliella salina (Volvocales, Chlorophyta) against experimentally induced fibrosarcoma on wistar rats. AB - The beta-carotene-yielding microalga, Dunaliella salina (Dunal) Teod. maintained in De Walne's medium was harvested and lyophilized. Fibrosarcoma was induced in rats by 20-methylcholanthrene. 0.5 g and 1.0 g of lyophilized D. salina powder was administered to the rats orally through carboxy methyl cellulose. Cisplatin was administered along with vitamin E to compare the protective effect of D. salina against fibrosarcoma. Administration of D. salina decreased the levels of cholesterol and lactate dehydrogenase as well as the activities of catalase, superoxide dismutase, serum aspartate aminotransaminase, serum alanine aminotransferase, when compared to control. A significant reduction in the levels of hepatic and renal RNA and DNA was observed in the sarcoma rats when treated with D. salina powder. Histopathological studies of tumor tissues showed regenerative and regressive changes. beta-carotene globules isolated from the powder of Dunaliella salina confirmed the presence of 9-cis-beta-carotene and all trans-beta-carotene. PMID- 16713217 TI - Reversal of multidrug resistance in cancer cells by Rhizoma Alismatis extract. AB - Prolonged chemotherapy may lead to the selective proliferation of multidrug resistant (MDR) cancer cells. In MDR HepG2-DR and K562-DR cells that over expressed P-glycoprotein (Pgp), the extract of the rhizomes of Alisma orientalis (Sam) Juzep. showed a synergistic growth inhibitory effect with cancer drugs that are Pgp substrates including actinomycin D, puromycin, paclitaxel, vinblastine and doxorubicin. At the same toxicity levels the herbal extract was more effective than verapamil, a standard Pgp inhibitor, in enhancing cellular doxorubicin accumulation and preventing the efflux of rhodamin-123 from the MDR cells. The extract restored the effect of vinblastine on the induction of G(2)/M arrest in MDR cells. Our data suggest that A. orientalis may contain components that are effective inhibitors of Pgp. PMID- 16713218 TI - Hypocholesteremic and antioxidant effects of Withania somnifera (Dunal) in hypercholesteremic rats. AB - Hypocholesteremic and antioxidant effects of Withania somnifera (WS) Dunal (Solanaceae) were investigated in hypercholesteremic male albino rats. When the root powder of WS was added to the diet at 0.75 and 1.5 gm/rat/day, hypercholesteremic animals registered significant decreases in total lipids ( 40.54%; -50.69%), cholesterol (-41.58%; -53.01%) and triglycerides (-31.25%; - 44.85%) in plasma. On the other hand, significant increases in plasma HDL cholesterol levels (+15.10%; +17.71%), HMG-CoA reductase activity (+19.51%; +26.02%) and bile acid content (+24.64%; +30.52%) of liver were noted in these animals. A similar trend was also noted in bile acid (+22.43%;+28.52%), cholesterol (+14.21%; +17.68%) and neutral sterol (+12.40%; +18.85%) excretion in the hypercholesteremic animals with WS administration. Further, a significant decrease in lipid-peroxidation (-35.29%; -36.52%) occurred in WS administered hypercholesteremic animals when compared to their normal counterparts. However, it appeared that WS root powder is also effective in normal subjects for decreasing lipid profiles. PMID- 16713219 TI - STW 5 (Iberogast) and its individual herbal components modulate intestinal electrophysiology of mice. AB - STW 5 (Iberogast), a phytomedicine agent consisting of a fixed combination of nine individual plant extracts, is widely used in the treatment of dyspepsia and motility related disorders. Little if anything is known on the possible influence on electrophysiological properties of intestinal smooth muscle by which STW 5 causes its beneficial effects. The aim of the present study was to investigate whether herbal extracts influence electrophysiological parameters of large and small intestine. For this purpose intracellular recordings of smooth muscle cell (SMC) of the circular muscle layer of different parts of mouse intestine were performed using standard microelectrode techniques. The resting membrane potential (RMP), excitatory and inhibitory neurotransmission in proximal colon, the frequency and the amplitude of slow waves in small intestine were investigated. The RMP of SMC was -46.4+/-3.8 mV, n=11 in the colon and -59+/-1.3 mV, n=15 in small intestine. STW 5 significantly depolarized the RMP of colonic (16.6+/-2.2 mV, n=6, p<0.05) and jejunal (9.6+/-1.6 mV, n=7, p<0.05) SMC. This depolarizing effect can be mainly attributed to the constituents of chamomile flower, Angelica root and greater celandine herb. Following the electrical field stimulations (EFSs), junction potentials are influenced in a distinct manner. Excitatory junctions potentials (EJPs) of the colon were not significantly reduced (13.1+/-4.8 vs. 10.1+/-2.8 n.s., n=6) but fast (fIJP) and slow (sIJP) inhibitory junction potentials of the murine colon are reduced significantly by STW 5 (fIJP: 21.6+/-8.1 vs. 11.6+/-2.1 and sIJP: 12.1+/-3.3 vs. 6.1+/-1.3 n=6, p<0.05). The basal frequency of small intestinal slow waves was 39.5+/-1.4 min( 1) and the amplitude was 23.1+/-0.9 mV, n=15. STW 5 significantly reduced amplitude and frequency of the slow waves (11.7+/-0.8 mV; 33.5+/-3.4 min(-1), n=6, p<0.05). This effect on slow waves represents the summation of effects of the nine individual phytoextracts. Whereas Angelica root and chamomile flower completely blocked the slow wave activity, bitter candy tuft increased the frequency and amplitude, greater celandine herb reduced frequency and amplitude of the slow wave, peppermint leaf reduced frequency and left amplitude unchanged and liquorice root, caraway fruit and lemon balm leaf had no effects in basic electrophysiological properties of SMC. This study demonstrates that STW 5 causes changes in SMC RMP, excitatory and inhibitory neurotransmission and slow wave rhythmicity. These effects represent a summation effect of different constituents of this phytotherapeuticum and prove that STW 5 has characteristic effects on intestinal electrophysiology. PMID- 16713220 TI - STW 5 (Iberogast) reduces afferent sensitivity in the rat small intestine. AB - INTRODUCTION: A limited number of drugs are available for the treatment of functional dyspepsia and irritable bowel syndrome. The efficacy of STW 5 (Iberogast) was previously shown in clinical trials. Since visceral hypersensitivity seems to be the prime pathomechanism of functional gastro intestinal disorders, the aim of this study was to explore whether STW 5 reduces intestinal afferent sensitivity in the upper gastrointestinal tract. METHODS: Two groups of male Wistar rats were pretreated with either the herbal preparation STW 5 or its vehicle (30.8% ethanol). Then, after 2h, general anesthesia was induced by pentobarbitone (60 mg kg(-1)i.p.) and extracellular multi-unit afferent recordings were obtained from mesenteric afferents innervating the proximal jejunum. The intestinal afferent nerve response to increasing doses of 5-HT and bradykinin were quantified as well as afferent discharge following a ramp distension of the adjacent intestinal loop from 0 to 60 cm H(2)O. RESULTS: Afferent discharge to 5-HT and bradykinin increased dose-dependently. Following the different doses of 5-HT, the peak in afferent nerve discharge was always reduced after pretreatment with STW 5 compared to controls with a response of 110+/-5 imp s(-1) after STW 5 and 128+/-3 in vehicle controls at the maximum dose (40 microg kg(-1); p<0.05; mean+/-SEM). For bradykinin, afferent responses were reduced following STW 5 at the 20 and 40 microg kg(-1) dose but not at 10 microg kg(-1) (40 microg kg(-1)176+/-7 imp s(-1) following STW 5 versus 200+/-6 imp s( 1) in controls; p<0.05). The ramp distension of the intestinal loop stimulated a rise in intestinal afferent nerve discharge that was always lower in the STW 5 pretreated group compared to vehicle controls with the exception of the discharge rate at the pressure level of 0 and 20 cm H(2)O (all other pressures up to 60 cm H(2)O p<0.05). CONCLUSIONS: Sensitivity of intestinal afferents to mechanical and chemical stimuli is reduced following treatment with the herbal preparation STW 5. This mechanism may help to explain why STW 5 relieves dyspeptic and bowel symptoms in patients. PMID- 16713222 TI - Antiproliferative activity and induction of apoptosis in human colon cancer cells treated in vitro with constituents of a product derived from Pistacia lentiscus L. var. chia. AB - In this report, we demonstrate that a 50% ethanol extract of the plant-derived product, Chios mastic gum (CMG), contains compounds which inhibit proliferation and induce death of HCT116 human colon cancer cells in vitro. CMG-treatment induces cell arrest at G(1), detachment of the cells from the substrate, activation of pro-caspases-8, -9 and -3, and causes several morphological changes typical of apoptosis in cell organelles. These events, furthermore, are time- and dose-dependent, but p53- and p21-independent. Apoptosis induction by CMG is not inhibited in HCT116 cell clones expressing high levels of the anti-apoptotic protein, Bcl-2, or dominant-negative FADD, thereby indicating that CMG induces cell death via a yet-to-be identified pathway, unrelated to the death receptor- and mitochondrion-dependent pathways. The findings presented here suggest that CMG (a) induces an anoikis form of cell death in HCT116 colon cancer cells that includes events associated with caspase-dependent pathways; and (b) might be developed into a chemotherapeutic agent for the treatment of human colon and other cancers. PMID- 16713223 TI - Antioxidative properties of the gastrointestinal phytopharmaceutical remedy STW 5 (Iberogast). AB - Since inflammation is a common mechanism of many gastrointestinal diseases, reactive oxygen metabolites may play an important role in their pathophysiology. Therefore it is interesting to know, whether phytopharmaceuticals known to modulate gastrointestinal motor function reveal also antioxidative properties. We tested STW 5 (Iberogast), its constituent nine different plant extracts, and some isolated compounds which are present in STW 5 for characterizing their antioxidative and radical quenching activities. The test assays consisted in pure chemical and complex cellular systems in which different types of reactive species were produced. Quantification of the effects was based on chemiluminescence reactions. The results show that all extracts contribute to the effect of the complete remedy STW 5, in the chemical systems in a strongly additive manner, in the cellular systems in a supraadditive manner. The largest contributions resulted from the extracts from peppermint and melissa leaves. Comparison of effects from isolated phytochemical compounds from the extracts with that of the extracts itself shows that usually the extract is more effective than the monosubstance which indicates also the synergism within the whole plant extracts. This means that the plant extracts present in STW 5 provide strong radical quenching activities that could also be involved in the therapeutic gastrointestinal actions. PMID- 16713224 TI - Uncontrolled asthma: a review of the prevalence, disease burden and options for treatment. AB - An estimated 300 million people are affected by asthma worldwide and the burden is likely to rise substantially in the next few decades. Estimates of the prevalence of asthma range from 7% in France and Germany to 11% in the USA and 15 18% in the United Kingdom. Approximately 20% of these patients have severe asthma, of which 20% is inadequately controlled. Patients with inadequately controlled severe persistent asthma are at a particularly high risk of exacerbations, hospitalization and death, and often have severely impaired quality of life. Current management of asthma focuses on a stepwise approach tailored to disease severity. In addition to needing high-dose inhaled corticosteroids (ICS) and long-acting beta(2)-agonists (LABAs), patients with severe persistent asthma often require additional controller medications, such as anti-leukotrienes, oral LABAs, oral corticosteroids and/or anti-IgE therapy. There is currently little evidence on which to base treatment decisions in patients with inadequately controlled severe persistent asthma already treated with ICS and LABAs. The anti-IgE monoclonal antibody omalizumab is the most recent addition to the list of treatment options for these patients and has been shown to reduce exacerbations and emergency visits and improve lung function, symptom scores and quality of life in patients with difficult-to-treat asthma whose symptoms remain inadequately controlled despite receiving ICS and LABAs. Comparative trials are needed to determine the merits of different treatments and strategies for patients with inadequately controlled severe persistent asthma and to identify patients likely to benefit from new treatment options. PMID- 16713225 TI - Longitudinal deteriorations in patient reported outcomes in patients with COPD. AB - Goals of effective management of patients with chronic obstructive pulmonary disease (COPD) include relieving their symptoms and improving their health status. We examined how such patient reported outcomes would change longitudinally in comparison to physiological outcomes in COPD. One hundred thirty-seven male outpatients with stable COPD were recruited for the study. The subjects health status was evaluated using the St. George's Respiratory Questionnaire (SGRQ) and the Chronic Respiratory Disease Questionnaire (CRQ). Their dyspnoea using the modified Medical Research Council (MRC) scale and their psychological status using the Hospital Anxiety and Depression Scale (HADS) were assessed upon entry and every 6 months thereafter over a 5-year period. Pulmonary function and exercise capacity as evaluated by peak oxygen uptake (VO2) on progressive cycle ergometry were also followed over the same time. Using mixed effects models to estimate the slopes for the changes, scores on the SGRQ, the CRQ, the MRC and the HADS worsened in a statistically significant manner over time. However, changes only weakly correlated with changes in forced expiratory volume in 1s (FEV(1)) and peak (VO2). We demonstrated that although changes in pulmonary function and exercise capacity are well known in patients with COPD, patient reported outcomes such as health status, dyspnoea and psychological status also deteriorated significantly over time. In addition, deteriorations in patient reported outcomes only weakly correlated to changes in physiological indices. To capture the overall deterioration of COPD from the subjective viewpoints of the patients, patient reported outcomes should be followed separately from physiological outcomes. PMID- 16713226 TI - Detection of changes in respiratory mechanics due to increasing degrees of airway obstruction in asthma by the forced oscillation technique. AB - Forced expiratory airflows and volumes are often used to assess the airway obstruction in asthmatics. However, forced maneuvers may change bronchial tone and modify airway patency. The aim of this study was to determine whether the Forced Oscillation Technique (FOT), which does not require forced manoeuvres, may be useful to describe the changes in respiratory mechanics in progressive asthma. This study involved 25 healthy and 84 asthmatics, including patients with normal spirometric exam (NE), mild moderate and severe obstruction. Resistive data were interpreted using the respiratory system resistance extrapolated at 0 Hz (R0), the mean respiratory resistance (Rm), and the resistance/frequency slope (S). Reactance data were interpreted by its mean values (Xm), the dynamic compliance (Crs,dyn), and resonant frequency (fr). Receiver operating characteristics curves were used to determine the sensitivity (Se) and specificity (Sp) of FOT parameters in identifying asthma. There were not statistically significant differences between the control and NE groups. Comparing the control and mild groups, significant increases of R0 (P<0.0007), Rm (P<0.003), and S (P<0.003) were observed. In reactive parameters, a significant reduction in Crs,dyn (P<0.04) was observed, while Xm and fr presented significant increases (P<0.0007 and P<0.006, respectively). Comparison between mild and moderate groups showed non-significant modifications in all of the parameters, except for Xm (P<0.02). In the late stages (moderate to severe obstruction), all of the resistive parameters, as well as the reactive ones Xm (P<0.007) and Crs,dyn (P<0.03), presented statistically significant modifications. Among the studied parameters, the effects of airway obstruction in asthma seem to be well described by R0, Rm, S and Xm, which were in close agreement with physiological fundamentals. The best parameters for detecting asthma were R0 (Se=81%, Sp=76%), S (Se=78%, Sp=72%) and Xm (Se=81%, Sp=80%). In conclusion, the results of this study suggest that the FOT can be proposed as an alternative method for the assessment of the respiratory mechanics in asthmatic patients, representing a promising solution to the problem of effort dependence. PMID- 16713227 TI - Nuclear receptor binding to the retinoic acid response elements of the phosphoenolpyruvate carboxykinase gene in vivo: effects of vitamin A deficiency. AB - Vitamin A deficiency decreases hepatic phosphoenolpyruvate carboxykinase (PEPCK) gene expression in mice and expression is restored with retinoic acid treatment in vivo. This report examines further the mechanism of retinoid regulation of the PEPCK gene in vivo. We have identified nuclear receptors that bind to retinoic acid response elements (RAREs) in the PEPCK promoter by electrophoretic mobility shift assay and have verified these in vivo using chromatin immunoprecipitation (ChIP) in mouse liver. Based on the results of our ChIP assay, hepatic nuclear factor (HNF)-4alpha, retinoid X receptor (RXR) alpha, retinoic acid receptor (RAR) alpha, peroxisome proliferator-activated receptor (PPAR) alpha and chicken ovalbumin upstream promoter transcription factor (COUP-TF) II bind to the downstream retinoic acid response unit RARE1/RARE2, and PPARalpha and RXRalpha bind to the upstream RARE3 of the PEPCK gene. HNF-4alpha, RXRalpha, RARalpha, PPARalpha and COUP-TFII bind PEPCK RAREs in a specific pattern that, with the exception of PPARalpha, does not change significantly with vitamin A deficiency. PPARalpha binding to the upstream retinoic acid response element is decreased in the vitamin A-deficient liver, when compared to the vitamin A-sufficient state. These results provide the first in vivo measures of nuclear receptor binding to the upstream and downstream RAREs of the PEPCK gene under conditions where the nucleosomal structure of the chromatin is maintained and the nuclear receptors are physically cross-linked in situ to the PEPCK DNA in intact liver. PMID- 16713228 TI - Hearts in adult offspring of copper-deficient dams exhibit decreased cytochrome c oxidase activity, increased mitochondrial hydrogen peroxide generation and enhanced formation of intracellular residual bodies. AB - The long-term effects of low dietary copper (Cu) intake during pregnancy and lactation on cardiac mitochondria in first-generation adult rats was examined. Rat dams were fed diets containing either low (1 mg/kg Cu) or adequate (6 mg/kg Cu) levels of dietary Cu beginning 3 weeks before conception and ending 3 weeks after birth. Cytochrome c oxidase (CCO) activity was 51% lower in isolated cardiac mitochondria from 21-day-old offspring of Cu-deficient dams than in the offspring of Cu-adequate dams. CCO activities in the cardiac mitochondria of 63- and 290-day-old offspring were 22% lower and 14% lower, respectively, in the offspring of Cu-deficient dams after they had been repleted with adequate dietary Cu from the time they were 21 days old. Electron micrographs showed that the size of residual bodies and the cellular volume they occupied in cardiomyocytes rose significantly between 63 and 290 days in the Cu-repleted offspring of Cu deficient dams, but not in the offspring of Cu-adequate dams. The rate of hydrogen peroxide generation by cardiac mitochondria also was 24% higher in the 290-day-old repleted offspring of Cu-deficient dams than in the offspring of Cu adequate dams. The increase in hydrogen peroxide production by cardiac mitochondria and in the relative volume and size of dense deposits in cardiomyocytes is consistent with increased oxidative stress and damage resulting from prolonged reduction of CCO activity in the offspring of Cu-deficient dams. PMID- 16713229 TI - Green tea catechins inhibit pancreatic phospholipase A(2) and intestinal absorption of lipids in ovariectomized rats. AB - This study was conducted to examine whether the inhibition of intestinal lipid absorption by green tea is associated with the inhibitory effect of its catechins on pancreatic phospholipase A(2) (PLA(2)). PLA(2) activity was assayed by using 1,2-dioleoylphosphatidylcholine (DOPC), porcine pancreatic PLA(2) and catechins at varying concentrations (0.075-1.80 micromol/L). The amount of 1-oleoyl-2 hydroxyphosphatidylcholine liberated was determined by HPLC. The percentage of inhibition of PLA(2) by catechins at 0.6 micromol increased in the order of (-) epicatechin (23.3%), (+)-catechin (CAT; 24.8%), (-)-epigallocatechin (25.7%), (-) epicatechin gallate (39.7%) and (-)-epigallocatechin gallate (EGCG; 64.9%). In an in vivo study, ovariectomized rats with lymph cannula were infused intraduodenally for 8 h with a triolein emulsion containing [dioleoyl-1-(14)C] phosphatidylcholine, DOPC, alpha-tocopherol (alphaTOH) and retinol (ROH) without (CAT0) or with CAT or EGCG. The lymphatic total (14)C-radioactivity was significantly lowered by EGCG (45.5+/-4.9% dose) compared with CAT (56.2+/-5.2% dose) and CAT0 (64.7+/-2.0% dose). The (14)C-radioactivity remaining in the small intestinal lumen and cecum was higher in EGCG (24.1% dose) than in CAT (9.5% dose) and CAT0 rats (9.0% dose). Significantly less (14)C radioactivity was incorporated into lymph triacylglycerol and cholesteryl ester in EGCG rats. The absorption of alphaTOH, used as a marker of extremely hydrophobic lipids, was significantly lower in EGCG (7.8+/-1.7 micromol) than in CAT (14.4+/-2.8 micromol) and CAT0 rats (16.8+/-2.1 micromol). The absorption of ROH was unaffected, whereas oleic acid output was lower in EGCG rats. The results show that EGCG inhibits the intestinal absorption of lipids, which is in part associated with its inhibition of phosphatidylcholine hydrolysis. Data suggest that EGCG may inhibit the absorption of other highly lipophilic organic compounds. PMID- 16713230 TI - Beneficial impact of crocetin, a carotenoid from saffron, on insulin sensitivity in fructose-fed rats. AB - Crocetin, a unique carotenoid with potent antioxidative and anti-inflammatory activities, is a major ingredient of saffron which is used as an important spice and food colorant in various parts of the world. In the present study, the effect of crocetin on insulin resistance and its related abnormalities induced by high fructose diet were investigated in male Wistar rats. Compared to the control rats fed on normal laboratory diet, fructose-fed rats developed a series of pathological changes including insulin resistance, hyperinsulinemia, dyslipidemia and hypertension. Although having no evident effect on the body weight, fructose feeding caused a marked increase in the weight of epididymal white adipose tissue. Furthermore, a significant reduction in the expression of both protein and mRNA of adiponectin (an insulin-sensitizing adipocytokine) was observed, whereas those of tumor necrosis factor (TNF)-alpha and leptin were enhanced in epididymal white adipose tissue in fructose-fed rats. These disorders were effectively normalized in crocetin-treated rats. Crocetin was also demonstrated here to alleviate free fatty acid (FFA)-induced insulin insensitivity and dysregulated mRNA expression of adiponectin, TNF-alpha and leptin in primary cultured rat adipocytes. These findings suggest the possibility of crocetin treatment as a preventive strategy of insulin resistance and related diseases. The favorable impact on adiponectin, TNF-alpha and leptin expression in white adipose tissue may be involved in the improvement of insulin sensitivity observed in crocetin-treated rats. PMID- 16713231 TI - Phospholipase D as a potential target for the antiproliferative effects of polyunsaturated fatty acids in rat thymocytes. AB - The effects of various saturated and unsaturated fatty acids (FAs) on the proliferative response and phospholipase D (PLD) activity of rat thymocytes were investigated. When added to culture medium as complexes with albumin, all the FAs tested, except stearic acid, inhibited the ConA-induced thymocyte proliferation, eicosapentaenoic (20:5n-3) and docosahexaenoic (22:6n-3) acids being the most inhibitory. Apart from 22:6n-3 which slightly increased the percentage of late apoptotic and necrotic thymocytes in the presence of mitogen, none of the FAs induced significant apoptosis or necrosis. A short 2-h preincubation of rat thymocytes in the presence of FA-albumin complexes was sufficient to induce a significant enrichment of cell phospholipids with each FA and to stimulate thymocyte PLD activity. However, 20:5n-3 was inactive despite a large enrichment in phospholipids. Furthermore, the PLD activity of activated thymocytes was negatively correlated to the proliferative response, with the exception of 20:5n 3-supplemented cells. These results support further our current hypothesis that PLD activity conveys antiproliferative signals in lymphoid cells, and suggest that 20:5n-3 inhibits thymocyte proliferation by a particular mechanism unrelated to that of the other FAs. PMID- 16713232 TI - Caloric restriction inhibits up-regulation of inflammatory cytokines and TNF alpha, and activates IL-10 and haptoglobin in the plasma of streptozotocin induced diabetic rats. AB - Diabetes mellitus is a significant risk factor for cardiovascular diseases, and low-grade systemic inflammation, mediated by oxidative stress, may play a central role. Caloric restriction (CR) has been reported to be effective in reducing oxidative stress during diabetes and moderating the expression of some markers of inflammation that are up-regulated during aging. Forty male Wistar rats were randomly divided into four groups: nondiabetic feeding ad libitum and under CR, and diabetic feeding ad libitum and under CR. The animals were subjected to 30% CR and ad libitum feeding for 9 weeks before the induction of diabetes by intraperitoneal injection with 35 mg/kg body weight streptozotocin. The inflammatory cytokines [interleukin (IL)-1beta, IL-4 and IL-6] and tumor necrosis factor alpha up-regulated in diabetes were found to be significantly depressed by CR, whereas the antiinflammatory mediators, haptoglobin and IL-10 levels, were increased. These results indicated that CR could prevent diabetic complications through suppression of inflammatory responses. PMID- 16713233 TI - Curcumin induces changes in expression of genes involved in cholesterol homeostasis. AB - Curcuminoids, the yellow pigments of curcuma, exhibit anticarcinogenic, antioxidative and hypocholesterolemic activities. To understand the molecular basis for the hypocholesterolemic effects, we examined the effects of curcumin on hepatic gene expression, using the human hepatoma cell line HepG2 as a model system. Curcumin treatment caused an up to sevenfold, concentration-dependent increase in LDL-receptor mRNA, whereas mRNAs of the genes encoding the sterol biosynthetic enzymes HMG CoA reductase and farnesyl diphosphate synthase were only slightly increased at high curcumin concentrations where cell viability was reduced. Expression of the regulatory SREBP genes was moderately increased, whereas mRNAs of the PPARalpha target genes CD36/fatty acid translocase and fatty acid binding protein 1 were down-regulated. LXRalpha expression and accumulation of mRNA of the LXRalpha target gene ABCg1 were increased at low curcumin concentrations. Although curcumin strongly inhibited alkaline phosphatase activity, an activation of a retinoic acid response element reporter employing secreted alkaline phosphatase was observed. These changes in gene expression are consistent with the proposed hypocholesterolemic effect of curcumin. PMID- 16713234 TI - Rice bran oil and oryzanol reduce plasma lipid and lipoprotein cholesterol concentrations and aortic cholesterol ester accumulation to a greater extent than ferulic acid in hypercholesterolemic hamsters. AB - Our laboratory has reported that the hypolipidemic effect of rice bran oil (RBO) is not entirely explained by its fatty acid composition. Because RBO has a greater content of the unsaponifiables, which also lower cholesterol compared to most vegetable oils, we wanted to know whether oryzanol or ferulic acid, two major unsaponifiables in RBO, has a greater cholesterol-lowering activity. Forty eight F(1)B Golden Syrian hamsters (Mesocricetus auratus) (BioBreeders, Watertown, MA) were group housed (three per cage) in cages with bedding in an air conditioned facility maintained on a 12-h light/dark cycle. The hamsters were fed a chow-based hypercholesterolemic diet (HCD) containing 10% coconut oil and 0.1% cholesterol for 2 weeks, at which time they were bled after an overnight fast (16 h) and segregated into 4 groups of 12 with similar plasma cholesterol concentrations. Group 1 (control) continued on the HCD, group 2 was fed the HCD containing 10% RBO in place of coconut oil, group 3 was fed the HCD plus 0.5% ferulic acid and group 4 was fed the HCD plus 0.5% oryzanol for an additional 10 weeks. After 10 weeks on the diets, plasma total cholesterol (TC) and non-high density lipoprotein cholesterol (HDL-C) (very low- and low-density lipoprotein) concentrations were significantly lower in the RBO (-64% and -70%, respectively), the ferulic acid (-22% and -24%, respectively) and the oryzanol (-70% and -77%, respectively) diets compared to control. Plasma TC and non-HDL-C concentrations were also significantly lower in the RBO (-53% and -61%, respectively) and oryzanol (-61% and -70%, respectively) diets compared to the ferulic acid. Compared to control and ferulic acid, plasma HDL-C concentrations were significantly higher in the RBO (10% and 20%, respectively) and oryzanol (13% and 24%, respectively) diets. The ferulic acid diet had significantly lower plasma HDL-C concentrations compared to the control (-9%). The RBO and oryzanol diets were significantly lower for plasma triglyceride concentrations compared to the control (-53% and -65%, respectively) and ferulic acid (-47% and -60%, respectively) diets. Hamsters fed the control and ferulic acid diets had significantly higher plasma vitamin E concentrations compared to the RBO (201% and 161%, respectively) and oryzanol (548% and 462%, respectively) diets; the ferulic acid and oryzanol diets had significantly lower plasma lipid hydroperoxide levels than the control (-57% and -46%, respectively) diet. The oryzanol-fed hamsters excreted significantly more coprostenol and cholesterol in their feces than the ferulic acid (127% and 120%, respectively) diet. The control diet had significantly greater aortic TC and FC accumulation compared to the RBO (115% and 89%, respectively), ferulic acid (48% and 58%, respectively) and the oryzanol (74% and 70%, respectively) diets. However, only the RBO and oryzanol diets had significantly lower aortic cholesterol ester accumulation compared to the control (-73% and -46%, respectively) diet. The present study suggests that at equal dietary levels, oryzanol has a greater effect on lowering plasma non-HDL C levels and raising plasma HDL-C than ferulic acid, possibly through a greater extent to increase fecal excretion of cholesterol and its metabolites. However, ferulic acid may have a greater antioxidant capacity via its ability to maintain serum vitamin E levels compared to RBO and oryzanol. Thus, both oryzanol and ferulic acid may exert similar antiatherogenic properties, but through different mechanisms. PMID- 16713235 TI - Changes in liver PPARalpha mRNA expression in response to two levels of high safflower-oil diets correlate with changes in adiposity and serum leptin in rats and mice. AB - The ligand-dependent transcription factor peroxisome proliferator-activated receptor alpha (PPARalpha) is known to be activated by common fatty acids and to regulate the expression of genes of various lipid oxidation pathways and transport. High-fat diets provide more fatty acids, which presumably could enhance lipid catabolism through up-regulation of PPARalpha signaling. However, high intake of fat could also lead to obesity. To examine PPARalpha signaling in high-fat feeding and obesity, this study examined the hepatic mRNA expression of PPARalpha and some of its target genes in Wistar rats and C57BL/6J mice fed two levels (20% or 30% wt/wt) of high-safflower-oil (SFO; oleic-acid-rich) diets until animals showed significantly higher body weight (13 weeks for rats and 22 weeks for mice) than those of control groups fed a 5% SFO diet. At the end of these respective feeding periods, only the rats fed 30% SFO and the mice fed 20% SFO among the two groups fed high-fat diets showed significantly higher body weight, white adipose tissue weight, serum leptin and mRNA expression of PPARalpha (P<.05) compared to the respective control groups. Despite elevated acyl-CoA (a PPARalpha target gene) protein and activity in both groups fed high fat diets, the mRNA expression level of most PPARalpha target genes examined correlated mainly to PPARalpha mRNA levels and not to fat intake or liver lipid levels. The observation that the liver PPARalpha mRNA expression in groups fed high-fat diets was significantly higher only in obese animals with elevated serum leptin implied that obesity and associated hyperleptinemia might have a stronger impact than dietary SFO intake per se on PPARalpha-regulated mRNA expression in the liver. PMID- 16713237 TI - Monitoring of odor compounds produced by solid waste treatment plants with diffusive samplers. AB - Nuisance caused by odors is one of the most important problems for waste management plants. To control an odor nuisance, it must first be quantified. The analytical difficulties in odor measurements are related to the high number of volatile components (belonging to several chemical classes), above all when the concentration is lower than the detection limit of the technique used for the measurement. In this work, 2-butanone, alpha-pinene, tetrachloroethylene, dimethyldisulfide, beta-pinene, limonene, phenol and benzoic acid are determined, because they are representative of some important classes of compounds with higher odor impact. The compounds are sampled with thermal desorbable radial diffusive samplers Radiello containing Tenax cartridges. The analytical repeatability and the complete thermal desorption of the cartridges were verified for each odor compound; the relative standard deviations for repeated samples and the recovery percentage were, respectively, less than 7% and about 97% for all compounds. The measurements of the linearity of sampling showed no systematic difference according to the collection period. The comparison between the odor threshold and the limit of detection demonstrated that this method is reliable for the recognition and quantification of odor compounds, allowing Public Administration to impose legal limits and the control agencies to verify them. PMID- 16713236 TI - Composition of source-sorted municipal organic waste collected in Danish cities. AB - Source-sorted municipal organic waste from different dwelling types in five Danish cities was sampled during one year. The samples were from permanent, full scale systems or temporary, experimental systems for collection of source-sorted municipal organic waste. Pre-treatment of the organic waste prior to biological treatment was used in all cities to remove foreign objects and provide size reduction. All sampling was performed after pre-treatment in order to obtain more homogeneous and representative samples. The sampling included both the pre treated waste and the reject from the pre-treatment allowing for estimation of the composition of the original waste. A total of 40 waste samples were chemically characterised with respect to 15 parameters. The waste generally consisted of around 88% VS of which an average of 80% was easily degradable. The average content of N, P and K in the dry matter of the organic waste was 2.5%, 0.4% and 0.9%, respectively. A general analysis of variance was applied to show the influence of the collection system, dwelling type and annual season on the waste composition. The content of plastic and crude fibres in the waste differed the most among the samples, probably due to use of different bag types (plastic and paper) in the different collection systems. Variations in the ash content and the calorific value might be explained by differences in the sorting instructions (whether soil and cat litter are allowed in the organic fraction). Significant seasonal variations were seen for ash, S and Cl. Dwelling type showed no statistically significant influence on any waste components. A test for uniform distribution of the p-values from the analysis of variance (Kolmogorov-Smirnov test) showed that the overall composition of the collected waste was strongly affected by the collection system (city) and season, while dwelling type had no significant influence. PMID- 16713238 TI - Assessment of medical wastes management practice: a case study of the northern part of Jordan. AB - This study includes a survey of the procedures available, techniques, and methods of handling and disposing of medical waste at medium (between 100 and 200 beds) to large (over 200 beds) size healthcare facilities located in Irbid city (a major city in the northern part of Jordan). A total of 14 healthcare facilities, including four hospitals and 10 clinical laboratories, serving a total population of about 1.5 million, were surveyed during the course of this research. This study took into consideration both the quantity and quality of the generated wastes to determine generation rates and physical properties. Results of the survey showed that healthcare facilities in Irbid city have less appropriate practices when it comes to the handling, storage, and disposal of wastes generated in comparison to the developed world. There are no defined methods for handling and disposal of these wastes, starting from the personnel responsible for collection through those who transport the wastes to the disposal site. Moreover, there are no specific regulations or guidelines for segregation or classification of these wastes. This means that wastes are mixed, for example, wastes coming from the kitchen with those generated by different departments. Also, more importantly, none of the sites surveyed could provide estimated quantities of waste generated by each department, based upon the known variables within the departments. Average generation rates of total medical wastes in the hospitals were estimated to be 6.10 kg/patient/day (3.49 kg/bed/day), 5.62 kg/patient/day (3.14 kg/bed/day), and 4.02 kg/patient/day (1.88 kg/bed/day) for public, maternity, and private hospitals, respectively. For medical laboratories, rates were found to be in the range of 0.053-0.065 kg/test-day for governmental laboratories, and 0.034-0.102 kg/test-day for private laboratories. Although, based on the type of waste, domestic or general waste makes up a large proportion of the waste volume, so that if such waste is not mixed with patient derived waste, it can be easily handled. However, based on infections, it is important for healthcare staff to take precautions in handling sharps and pathological wastes, which comprises only about 26% of the total infectious wastes. Statistical analysis was conducted to develop mathematical models to aid in the prediction of waste quantities generated by the hospitals studied, or similar sites in the city that are not included in this study. In these models, the number of patients, number of beds, and hospital type were determined to be significant factors on waste generation. Such models provide decision makers with tools to better manage their medical waste, given the dynamic conditions of their healthcare facilities. PMID- 16713239 TI - Municipal solid waste disposal in Portugal. AB - In recent years municipal solid waste (MSW) disposal has been one of the most important environmental problems for all of the Portuguese regions. The basic principles of MSW management in Portugal are: (1) prevention or reduction, (2) reuse, (3) recovery (e.g., recycling, incineration with heat recovery), and (4) polluter-pay principle. A brief history of legislative trends in waste management is provided herein as background for current waste management and recycling activities. The paper also presents and discusses the municipal solid waste management in Portugal and is based primarily on a national inquiry carried out in 2003 and directed to the MSW management entities. Additionally, the MSW responsibility and management structure in Portugal is presented, together with the present situation of production, collection, recycling, treatment and elimination of MSW. Results showed that 96% of MSW was collected mixed (4% was separately collected) and that 68% was disposed of in landfill, 21% was incinerated at waste-to-energy plants, 8% was treated at organic waste recovery plants and 3% was delivered to sorting. The average generation rate of MSW was 1.32 kg/capita/day. PMID- 16713240 TI - Gray correlation analysis and prediction models of living refuse generation in Shanghai city. AB - A better understanding of the factors that affect the generation of municipal living refuse (MLF) and the accurate prediction of its generation are crucial for municipal planning projects and city management. Up to now, most of the design efforts have been based on a rough prediction of MLF without any actual support. In this paper, based on published data of socioeconomic variables and MLF generation from 1990 to 2003 in the city of Shanghai, the main factors that affect MLF generation have been quantitatively studied using the method of gray correlation coefficient. Several gray models, such as GM(1,1), GIM(1), GPPM(1) and GLPM(1), have been studied, and predicted results are verified with subsequent residual test. Results show that, among the selected seven factors, consumption of gas, water and electricity are the largest three factors affecting MLF generation, and GLPM(1) is the optimized model to predict MLF generation. Through this model, the predicted MLF generation in 2010 in Shanghai will be 7.65 million tons. The methods and results developed in this paper can provide valuable information for MLF management and related municipal planning projects. PMID- 16713241 TI - Recombinant human voltage-gated skeletal muscle sodium channels are pharmacologically functional in planar lipid bilayers. AB - Human voltage-gated sodium ion channels are major sites of action for drugs and toxins that modulate cellular excitability, and are therefore key molecular targets for ion channel research, high throughput screening for new drugs, and toxin detection. Protein suitable for these applications must be produced in a functionally active form. We report the successful use of ion metal affinity chromatography (IMAC) to purify C-terminal polyhistidine tagged human skeletal muscle voltage-gated sodium (hSkM1-HT) channels from Sf9 insect cells; hSkM1 channels were pharmacologically functional when reconstituted into liposomes and incorporated into planar bilayer lipid membranes. hSkM1-HT single channel currents activated by veratridine had a conductance of 21 pS and those activated by brevetoxin, 16 pS. Channel activity was inhibited by tetrodotoxin and saxitoxin. This protein is suitable for the development of biosensor and high throughput screening technologies. PMID- 16713242 TI - Influence of the first amplifier stage in MEA systems on extracellular signal shapes. AB - Recording of extracellular signals with planar metal microelectrodes (ME) has already been presented more than 30 years ago. To date, microelectrode array (MEA) systems are able to measure extracellular signals at about 64 sites, simultaneously. This enables monitoring of electrical activity of many cells in a large area. The extracellular recording technique has become a widely used method for neurological, toxicological or pharmacological studies. It already proved its potential to supplement the classical methods in electrophysiology. The interpretation of the recorded signal shapes in order to extract electrophysiological meaningful data--however--is still under discussion. In this article, we analyse the preamplifier circuit for extracellular recording of cardiac myocyte signals. We use a circuit model for the cell-electrode contact including the first amplification stage. In test experiments, we observe different signal shapes, when different shunt resistors are introduced at the input of the preamplifier. According to the frequency spectra of the recordings, we evaluate the transfer function between the source signal and the readout signal. As a result of our studies, an optimum readout electronics for originally, preserved extracellular signal shapes is proposed. Our amplifier design will be most valuable, if the use of small microelectrodes with high input impedances for in vitro as well as for in vivo experiments is desired. PMID- 16713243 TI - Engineering yeasts for xylose metabolism. AB - Technologies for the production of alternative fuels are receiving increased attention owing to concerns over the rising cost of petrol and global warming. One such technology under development is the use of yeasts for the commercial fermentation of xylose to ethanol. Several approaches have been employed to engineer xylose metabolism. These involve modeling, flux analysis, and expression analysis followed by the targeted deletion or altered expression of key genes. Expression analysis is increasingly being used to target rate-limiting steps. Quantitative metabolic models have also proved extremely useful: they can be calculated from stoichiometric balances or inferred from the labeling of intermediate metabolites. The recent determination of the genome sequence for P. stipitis is important, as its genome characteristics and regulatory patterns could serve as guides for further development in this natural xylose-fermenting yeast or in engineered Saccharomyces cerevisiae. Lastly, strain selection through mutagenesis, adaptive evolution or from nature can also be employed to further improve activity. PMID- 16713244 TI - Regulation of Ca2+-permeable AMPA receptors: synaptic plasticity and beyond. AB - AMPA-type glutamate receptors (AMPARs) mediate most fast excitatory synaptic transmission in the brain. Diversity in excitatory signalling arises, in part, from functional differences among AMPAR subtypes. Although the rapid insertion or deletion of AMPARs is recognised as important for the expression of conventional forms of long-term synaptic plasticity--triggered, for example, by Ca2+ entry through NMDA-type glutamate receptors--only recently has attention focused on novel forms of plasticity that are regulated by, or alter the expression of, Ca2+ permeable AMPARs. The dynamic regulation of these receptors is important for normal synaptic function and in disease states. PMID- 16713245 TI - Mechanisms of neural plasticity following brain injury. AB - Brain insults cause rapid cell death, and a disruption of functional circuits, in the affected regions. As the injured tissue recovers from events associated with cell death, regenerative processes are activated that over months lead to a certain degree of functional recovery. Factors produced by new neurons and glia, axonal sprouting of surviving neurons, and new synapse formation help to re establish some of the lost functions. The timing and location of such events is crucial in the success of the regenerative process. Comprehensive gene expression profiling and proteomic analyses have enabled a deeper molecular and cellular mechanistic understanding of post-injury brain regeneration. These new mechanistic insights are aiding the design of novel therapeutic modalities that enhance regeneration. PMID- 16713246 TI - Novel presynaptic mechanisms for coincidence detection in synaptic plasticity. AB - Long-term plasticity typically relies on postsynaptic NMDA receptors to detect the coincidence of pre- and postsynaptic activity. Recent studies, however, have revealed forms of plasticity that depend on coincidence detection by presynaptic NMDA receptors. In the amygdala, cortical afferent associative presynaptic long term potentiation (LTP) requires activation of presynaptic NMDA receptors by simultaneous thalamic and cortical afferents. Surprisingly, both types of afferent can also undergo postsynaptically induced NMDA-receptor-dependent LTP. In the neocortex, spike-timing-dependent long-term depression (LTD) requires simultaneous activation of presynaptic NMDA autoreceptors and retrograde signalling by endocannabinoids. In cerebellar LTD, presynaptic NMDA receptor activation suggests that similar presynaptic mechanisms may exist. Recent studies also indicate the existence of presynaptic coincidence detection that is independent of NMDA receptors, suggesting that such mechanisms have a widespread role in plasticity. PMID- 16713247 TI - The types and prevalence of alternative splice forms. AB - The finding that eukaryotic gene structures are extremely complex prompted the development of new experimental techniques for the accurate measurement of transcription start site usage and of the expression of alternative splice forms. On the computational side, analyses of large databases of splice variants revealed differences in the length, motif composition and selection pressure between constitutive and alternatively spliced exons. Such features are being incorporated into novel computational tools for gene structure prediction. The result of these investigations is a continuously improving catalogue of alternative splice forms. How the expression of these alternative splice forms is regulated remains one of the major open questions. PMID- 16713248 TI - DNA structures, repeat expansions and human hereditary disorders. AB - Expansions of simple DNA repeats are responsible for more than two dozen hereditary disorders in humans, including fragile X syndrome, myotonic dystrophy, Huntington's disease, various spinocerebellar ataxias, Friedreich's ataxia and others. During the past decade, it became clear that unusual structural features of expandable repeats greatly contribute to their instability and could lead to their expansion. Furthermore, DNA replication, repair and recombination are implicated in the formation of repeat expansions, as shown in various experimental systems. The replication model of repeat expansion stipulates that unusual structures of expandable repeats stall replication fork progression, whereas extra repeats are added during replication fork restart. It also explains the bias toward repeat expansion or contraction that was observed in different organisms. PMID- 16713250 TI - Structure and function of telomerase RNA. AB - Maintenance of telomeres by the enzyme telomerase is essential for genomic stability and cell viability in ciliates, vertebrates and yeast. The minimal components of telomerase required for catalytic activity are the telomerase reverse transcriptase (TERT) protein and the template-containing telomerase RNA (TER). Recent studies have afforded significant advances in the biophysical characterization of telomerase RNAs from various species. The first TER structures have been reported, for regions of the catalytically essential pseudoknot and CR4/CR5 domains of human TER, and provide a structural basis for interpretation of mutational and biochemical data. The domains and interactions of the Tetrahymena thermophila telomerase holoenzyme RNA and protein components have been further characterized biochemically, and structures of the TER template boundary element and the N-terminal domain of T. thermophila TERT have been determined. Phylogenetic and biochemical analyses of yeast TERs have revealed core structural elements in common with ciliates and vertebrates, and the minimal domains required for function in vivo. PMID- 16713251 TI - Target selection for complex structural genomics. AB - Most cellular processes are carried out by macromolecular assemblies and regulated through a complex network of transient protein-protein interactions. Genome-wide interaction discovery experiments are already delivering the first drafts of whole organism interactomes and, thus, depicting the limits of the interaction space. However, a complete understanding of molecular interactions can only come from high-resolution three-dimensional structures, as they provide key atomic details about the binding interfaces. The launch of structural genomics initiatives focused on protein interactions and complexes could quickly fill up the interaction space with structural details, offering a new perspective on how cell networks operate at atomic level. Clear target selection strategies that rationally identify the key interactions and complexes that should be first tackled are fundamental to maximize the return, minimize the costs and prevent experimental difficulties. PMID- 16713252 TI - Microbial metabolism of methyl protodioscin by Aspergillus niger culture--a new androstenedione producing way from steroid. AB - Methyl protodioscin (1), a natural furostanol biglycoside steroid, was a preclinical anticancer drug, which showed potent activity against most cell lines from leukemia and solid tumors in the National Cancer Institute's (NCI) human cancer panel. Metabolism of methyl protodioscin by Aspergillus niger was investigated. Seven metabolites were isolated and identified. Two main metabolites were pregnane glycosides and four were furostanol glycosides, together with the aglycone. It was found that steroidal saponin skeleton could be converted to pregnenolone skeleton only using microbial methods, which must have chemical procedures in the reported literatures. The proposed biosynthetic pathways of the microbial conversion products of methyl protodioscin were drawn. The found enriched the reaction types of microbial bioconversion and provided a new producing way of androstenedione from steroid. Most metabolites showed strong cytotoxic activities against HepG2, NCI-H460, HeLa, and MCF-7 cell lines. PMID- 16713253 TI - Decreased responsiveness of naturally occurring mutants of human estrogen receptor alpha to estrogens and antiestrogens. AB - Estrogen receptor alpha (ERalpha) is a ligand-inducible transcription factor that mediates the biological effects of estrogens and antiestrogens. Many point mutations in the human ERalpha gene have been reported to be associated with breast cancer, endometrial cancer, and psychiatric diseases. However, functional analyses for most mutants with amino acid changes are still lacking. In the present study, to investigate the effects of point mutations on the function, gel shift assays and luciferase assays were performed for eight kinds of mutated ERalpha proteins, including a single nucleotide change of C207G (N69K), G478T (G160C), T887C (L296P), A908G (K303R), C926T (S309F), A1058T (E353V), A1186G (M396V), and G1231deletion (411fsX7). The mutated ERalpha expression plasmids were constructed by site-directed mutagenesis. With gel-shift assays using in vitro translated ERalpha proteins, binding to the consensus estrogen response element (ERE) was observed for the mutated ERalpha proteins except ERalpha (G160C) and ERalpha (411fsX7), the binding of which was comparable with that of the wild type. Western blot analyses showed that ERalpha (G160C) could not be efficiently translated with the in vitro transcription/translation system and that ERalpha (411fsX7) produced a truncated protein. To investigate the transactivation potency, wild-type or mutated ERalpha expression plasmids were co transfected with pGL3-3EREc38 reporter plasmid into human breast adenocarcinoma MDA-MB-435 cells. The concentration-response curves (10pM-100nM E2) of the mutant ERalpha proteins except ERalpha (E353V) and ERalpha (411fsX7) were similar to that of wild-type ERalpha. However, at a low level of E2 (100pM), the mutants ERalpha (N69K), ERalpha (L296P), ERalpha (S309F), and ERalpha (M396V) showed a significant decrease of transactivation compared with that of the wild-type ERalpha. The mutants ERalpha (E353V) and ERalpha (411fsX7) did not show responsiveness to E2 and antiestrogens, 4-hydroxytamoxifen (4OHT) and ICI 182,780. The mutant ERalpha (S309F) showed decreased responsiveness for the antiestrogenicity of 4OHT. In conclusion, we found that some of the naturally occurring human ERalpha mutants with amino acid changes may have an altered responsiveness to estrogen and antiestrogens. PMID- 16713254 TI - The native anti-glucocorticoid paradigm. AB - Circulating 3beta-hydroxysteroids including dehydroepiandrosterone (DHEA) are 7alpha-hydroxylated by the cytochrome P450-7B1 in the liver, skin and brain, which are the target organs of glucocorticoids. Anti-glucocorticoid effects with 7alpha-hydroxy-DHEA were observed in vivo without an interference with glucocorticoid binding to its receptor. In the organs mentioned above, the circulating inactive cortisone was reduced into active cortisol by the 11beta hydroxysteroid dehydrogenase type 1 (11beta-HSD1). We demonstrated that 7alpha hydroxy-DHEA was also a substrate for this enzyme. Studies of the 11beta-HSD1 action on 7alpha-hydroxy-DHEA showed the reversible production of 7beta-hydroxy DHEA through an intermediary 7-oxo-DHEA, and the kinetic parameters favored this production over that of active glucocorticoids. Both the production of 7alpha hydroxysteroids and their interference with the activation of cortisone into cortisol are basic to the concept of native anti-glucocorticoids efficient at their production site. This opens a promising new area for research. PMID- 16713256 TI - Alpha-galactosidase production by Aspergillus oryzae in solid-state fermentation. AB - Comparisons were made for alpha-galactosidase production using red gram plant waste (RGPW) with wheat bran (WB) and other locally available substrates using the fungus Aspergillus oryzae under solid-state fermentation (SSF). RGPW proved to be potential substrate for alpha-galactosidase production as it gave higher enzyme titers (3.4 U/g) compared to WB (2.7 U/g) and other substrates tested. Mixing WB with RGPW (1:1, w/w) resulted enhanced alpha-galactosidase yield. The volume of moistening agent in the ratio of 1:2 (w/v), pH 5.5 and 1 ml (1 x 10(6) spores) of inoculum volume and four days incubation were optimum for alpha galactosidase production. Increase in substrate concentration (RGPW+WB) did not decrease enzyme yield in trays. PMID- 16713255 TI - Nutrient recovery from domestic wastewater using a UASB-duckweed ponds system. AB - The pilot-scale wastewater treatment system used in this study comprised a 40-l UASB reactor (6-h HRT) followed by three duckweed ponds in series (total HRT 15 days). During the warm season, the treatment system achieved removal values of 93%, 96% and 91% for COD, BOD and TSS, respectively. Residual values of ammonia, TKN and total phosphorus were 0.41 mg N/l, 4.4 mg N/l and 1.11 mg P/l, with removal efficiencies of 98%, 85% and 78%, respectively. The system achieved 99.998% faecal coliform removal during the warm season with final effluent containing 4 x 10(3) cfu/100 ml. During the winter, the system was efficient in removing COD, BOD and TSS but not nutrients. The system was deficient in the removal of faecal coliforms during the winter, producing effluent with 4.7 x 10(5) cfu/100 ml. During the warm season, the N removal consisted of 80% by plant uptake, 5% by sedimentation and 15% unaccounted for. A duckweed production rate of 33 t dry matter per hectare per 8 months was achieved. PMID- 16713257 TI - Shaken not stirred: a facile synthesis of 1,4-bis(furo[2,3-d]-pyrimidine 2,4(1H,3H)-dione-5-yl)benzenes by one-pot reaction of isocyanides, N,N' dimethylbarbituric acid, and terephthaldialdehyde. AB - A simple and efficient synthesis of 1,4-bis(furo[2,3-d]pyrimidine-2,4(1H,3H) dione-5-yl)benzene derivatives was achieved via a one-pot three-component reaction of isocyanides, N,N'-dimethylbarbituric acid, and terephthaldialdehyde in DMF at room temperature for 30 min. These improved reaction conditions allow the preparation of highly substituted furopyrimidinones in high yields and purity under mild reaction conditions. PMID- 16713258 TI - Photocleavage of o-nitrobenzyl ether derivatives for rapid biomedical release applications. AB - The externally controlled cleavage of covalently linked prodrugs, proteins, or solid-phase formulation vehicles offers potential advantages for controlled drug or gene delivery. A series of o-nitrobenzyl ester compounds (1-8) were synthesized to allow a systematic study of photolability. The o-nitrobenzyl ester was strictly required for photolability, while imido esters were not photolabile. The degradation kinetics of 1-o-phenylethyl ester was an order of magnitude faster than that of o-nitrobenzyl ester. Tosylate, phosphate, and benzoate derivatives of 1-o-nitrophenylethyl displayed similar photolability (>80% decomposition within 10 min at 3.5 mW/cm2 at 365 nm). O-o-Nitrobenzyl O',O'' diethyl phosphate displayed the fastest decomposition at photoirradiation condition (3.5 mW/cm2, 365 nm) suitable for biological systems. We report the synthesis and photo-decomposition of 1-o-nitrophenylethyl derivatives amenable for the creation of photolabile prodrugs or formulation particles for drug depots, DNA condensation, or tissue engineering applications. PMID- 16713259 TI - Substituted N-(2-aminophenyl)-benzamides, (E)-N-(2-aminophenyl)-acrylamides and their analogues: novel classes of histone deacetylase inhibitors. AB - Inhibition of histone deacetylases (HDACs) is emerging as a new strategy in human cancer therapy. Novel 2-aminophenyl benzamides and acrylamides, that can inhibit human HDAC enzymes and induce hyperacetylation of histones in human cancer cells, have been designed and synthesized. These compounds selectively inhibit proliferation and cause cell cycle arrest in various human cancer cells but not in normal cells. The growth inhibition of 2-aminophenyl benzamides and acrylamides against human cancer cells in vitro is reversible and is dependent on the induction of histone acetylation. Compounds of this class can significantly reduce tumor growth in human tumor xenograft models. PMID- 16713261 TI - Identification of metabolites of the tryptase inhibitor CRA-9249: observation of a metabolite derived from an unexpected hydroxylation pathway. AB - The metabolites of the tryptase inhibitor CRA-9249 were identified after exposure to liver microsomes. CRA-9249 was found to be degraded rapidly in liver microsomes from rabbit, dog, cynomolgus monkey, and human, and less rapidly in microsomes from rat. The key metabolites included cleavage of an aryl ether, in addition to an unexpected hydroxylation of the amide side chain adjacent to the amide nitrogen. The chemical structures of both metabolites were confirmed by synthesis and comparison to material isolated from the liver microsomes. Several suspected hydroxylated metabolites were also synthesized and analyzed as part of the structure identification process. PMID- 16713260 TI - Anti-HIV activity of stilbene-related heterocyclic compounds. AB - Viral transcription has not been routinely targeted in the development of new antiviral drugs. This crucial step of the viral cycle depends on the concerted action of cellular and viral proteins such as NF-kappaB and Tat. In the present study, stilbene-related heterocyclic compounds including benzalphthalide, phthalazinone, imidazoindole and pyrimidoisoindole derivatives are tested for their anti-HIV activity. Original assays based on recombinant viruses were used to evaluate HIV replication inhibition and stably transfected cell lines were used to evaluate inhibition of Tat and NF-kappaB proteins. Some of the stilbene related heterocyclic compounds analysed displayed anti-HIV activity through interference with NF-kappaB and Tat function. Moreover, compounds inhibiting both targets displayed a stronger activity on viral replication. PMID- 16713262 TI - Proton dissociation is important to understanding structure-activity relationships of gallic acid antioxidants. AB - Gallic acid derivatives (GADs) can efficiently scavenge free radicals, which is partially responsible for their neuroprotective effects. As GADs tend to deprotonate to give birth to GAD anions, which has big influence on the radical scavenging behaviors of GADs, to understand the structure-activity relationships (SARs) of GAD antioxidants, the anions should be taken into consideration. In this paper, a combined density functional theory method, labeled as (RO)B3LYP/6 311+G(2d,2p)//AM1/AM1, was employed to calculate homolytic O-H bond dissociation enthalpies and adiabatic ionization potentials for GADs and derived anions in solvent (ethanol), by which the experimentally observed SARs of GADs were better elucidated. PMID- 16713263 TI - The influence of pH on the G-quadruplex binding selectivity of perylene derivatives. AB - Three new perylene derivatives with branched ionizable side chains were synthesized, and their G-quadruplex binding specificities were compared by spectroscopic and electrophoretic analysis with two well-studied G-quadruplex ligands: PIPER and TmPyP4. The value of pH and consequent charge formation and self-aggregation of these perylene derivatives influences not only the type of G quadruplex formation, but also the G-quadruplex binding selectivity. PMID- 16713264 TI - Dibasic non-imidazole histamine H3 receptor antagonists with a rigid biphenyl scaffold. AB - A class of rigid, dibasic, non-imidazole H3 antagonists was developed, starting from a series of previously described flexible compounds. The original polymethylene chain between two tertiary amine groups was replaced by a rigid scaffold, composed by a phenyl ring or a biphenyl fragment. Modulation of the distance between the two amine groups, and of their alkyl substituents, was driven by superposition of molecular models and docking into a receptor model, resulting in the identification of 1,1'-[biphenyl-4,4'-diylbis(methylene)]bis piperidine (5) as a subtype-selective H3 antagonist with high binding affinity (pKi=9.47) at human H3 histamine receptor. PMID- 16713265 TI - 5-benzylidene-hydantoins as new EGFR inhibitors with antiproliferative activity. AB - A series of 1,5-disubstituted hydantoins, whose structure was designed to interact at the ATP-binding site of EGFR, was synthesized and evaluated for inhibition of EGFR kinase activity and antiproliferative action. Some of these compounds, characterized by a 1-phenethyl and a 5-(E)-benzylidene substituent, inhibited EGFR autophosphorylation and polyGAT phosphorylation, and also inhibited the growth and proliferation of human A431 cells, which overexpress EGFR. These compounds can therefore be regarded as examples of a new scaffold for tyrosine kinase inhibitors. PMID- 16713266 TI - Glutathione S-transferase polymorphisms associated with risk of breast cancer in southern Taiwan. AB - In this study, the genetic polymorphisms associated with breast cancer in southern Taiwan were investigated. Two categories of genes were analyzed: (1) BRCA1, BRCA2, and Rad51, the DNA repair factors involved in homologous recombinational repair; and (2) CYP1A1, COMT, GST, and NAT2, the xenobiotic metabolizing enzymes (XME) involved in estrogen metabolism. We found that the number of deletions and/or mutations in the GST genes was highly correlated with the occurrence of breast cancer. These data suggest that the GST enzymes, which detoxify the catechol estrogen quinones, are important target molecules for screening in populations at high risk of breast cancer. PMID- 16713267 TI - Self-reported infection control practices and perceptions of HIV/AIDS risk amongst emergency department nurses in Botswana. AB - This descriptive exploratory study investigated the reported practices and perceptions of emergency nurses related to infection control in the context of the Human Immunodeficiency Virus/Acquired Immune Deficiency Syndrome (HIV/AIDS) pandemic in Botswana. Quantitative and qualitative data were collected using a self-administered questionnaire. Forty questionnaires were distributed to nurses with emergency department experience in Botswana, with a response rate of 55% (n = 22). Quantitative data were analysed using descriptive statistics while qualitative data were subjected to thematic and content analysis. The majority of respondents reported compliance with universal precautions at the hospital emergency department. However, qualitative data highlighted resource constraints that may hinder compliance with universal precautions such as a lack of appropriate facilities, a shortage of equipment and materials, inadequate staffing and absence of sustainable in-service education programs. Further, the reported compliance with Universal Precautions had not removed the fear of exposure to HIV/AIDS and perceived risk of transmission to family. The authors recommend in-service education and practice initiatives to promote sustainable compliance with universal precautions and realistic risk perception among nurses. Further research is required to evaluate nurses' compliance with universal precautions in developing countries using observational methods or in-depth interviews. This would enable exploration of nurses' actions regarding compliance with universal precautions. PMID- 16713268 TI - Thrombomodulin induction in cultured human endothelial cells by 9-cis-locked retinoic acid analogues. AB - 9-cis-Retinoic acid (RA) analogues devised to lock the 9-cis double bond by ring formation were synthesized using two stereoselective carbon-carbon bond formation reactions as key steps. The palladium-mediated Suzuki reaction was adopted to construct a 7E-double bond (RA numbering) and the Horner-Emmons olefination was employed for stereoselective 11E-double bond (RA numbering) formation. The synthesized 9-cis-RA analogues that are locked by five-membered ring systems (cyclopentene, dihydrofuran, and dihydrothiophene) were shown to have comparable thrombomodulin induction activities to that of 9-cis RA. Conformational analysis of these compounds showed their similarity to 9-cis RA in the spatial orientation of the side chain and the terminal carboxy group. PMID- 16713269 TI - Anticancer activities of some newly synthesized pyridine, pyrane, and pyrimidine derivatives. AB - A series of pyridine, pyrane, and pyrimidine derivatives (2-11) were newly synthesized using nitrobenzosuberone 1 as a starting material. The antitumor activities of the synthesized compounds were evaluated utilizing 59 different human tumor cell lines, representing leukemia, melanoma, lung, colon, brain, ovary, breast, prostate as well as kidney. Some of the tested compounds especially 2, 3, 4c, 6, 7, 9b, 10a, and 11 exhibited better in vitro antitumor activities at low concentration (log(10) GI(50) = -4.7) against the used human tumor cell lines. Additionally, compounds 3, 4c, 6, 7, and 9b were highly selective to inhibit leukemia cell lines. The detailed synthesis, spectroscopic data and antitumor properties for the synthesized compounds were reported. PMID- 16713270 TI - Synthesis and evaluation of acridine- and acridone-based anti-herpes agents with topoisomerase activity. AB - The discovery of new non-nucleoside antiviral compounds is of significant and growing interest for treating herpes virus infections due to the emergence of nucleoside-resistant strains. Using a whole cell virus-induced cytopathogenic assay, we tested a series of substituted triaryl heterocyclic compounds including acridones, xanthones, and acridines. The compounds which showed activity against Herpes Simplex-1 and/or Herpes Simplex-2 were further assayed for inhibition of topoisomerase activity to gain insight into the mechanism of action. The results indicate that the acridine analogs bearing substituted carboxamides and bulky 9 amino functionalities are able to inhibit herpes infections as well as inhibit topoisomerase II relaxation of supercoiled DNA. Given the mechanism of action of amsacrine (a closely related, well-studied 9-amino substituted acridine), the compounds were further tested in a DNA topoisomerase II cleavage assay to determine if the compounds function as poisons. The results show that the acridines synthesized in this study function through a different mechanism to that of amsacrine, most likely by blocking topoisomerase binding to DNA (akin to that of aclarubicin). This not only suggests a unique mechanism of action in treating herpes virus infections, but also may be of great interest in the development of anticancer agents that target topoisomerase II activity. PMID- 16713271 TI - Regulatory role of kinases and phosphatases on the internalisation of caveolae in HepG2 cells. AB - The caveolar cycle is thought to be regulated by synchronised function of kinases and phosphatases. Using ocadaic acid--a serine/threonine protein phosphatase inhibitor--and an inhibitor of tyrosine phosphatase (sodium orthovanadate) we have followed the internalisation of caveolae. Since albumin binding to its receptor (gp60) can induce pinching off of caveolae from the plasma membrane, we also used this physiological ligand to induce the internalisation. Our confocal microscopic results show that both ocadaic acid and vanadate treatments have significantly decreased caveolin (caveolin-1 and -2) labelling on the cell surface, while the cytoplasmic labelling became much stronger. Quite often large, strongly labelled "granules" appear at the perinuclear region. Very strong caveolin labelling was detected along the actin-cytoskeleton suggesting that caveolae might move along these filaments. Our electron microscopic results also show an intensive caveolae pinching off from the plasma membrane. After ocadaic acid and vanadate treatments the number of surface connected vesicles (caveolae) decreases. At the same time, large multivesicular bodies (termed caveosomes) appear in the perinuclear area of the cytoplasm. By immunoprecipitation and Western blot analysis we detect an increased tyrosine phosphorylation of a approximately 29kDa protein in ocadaic acid and vanadate treated samples. This protein was identified as caveolin-2. No significant change in the tyrosine phosphorylation of caveolin-1 was found. From these data we can conclude that caveolae internalisation is regulated by phosphorylation of caveolin-2. PMID- 16713272 TI - Ultrastructure of the skin melanophores and iridophores in paddlefish, Polyodon spathula. AB - The ultrastructure of melanophores and iridophores of Polyodon spathula has been examined by transmission electron microscopy. In the skin, two types of chromatophores, melanophores and iridophores were founded. Melanophores were localized both in epidermis and dermis. Epidermal melanophores were present on the dorsal region of the trunk, sides, outer surface of the operculum and rostrum. Iridophores were founded in the dermis from ventral skin. The cytoplasm of iridophores is filled with reflecting platelets with variable orientation. The length of the long axis of the platelets varies from 1 to 2.10 microm. PMID- 16713273 TI - An experiment using neutron activation analysis and a rare earth element to mark cotton plants and two insects that feed on them. AB - Studies on insect dispersal and other behaviors can benefit from using markers that will not alter flight and fitness. Rare earth elements, such as samarium (Sm), have been used as ingested markers of some insects and detected using neutron activation analysis (NAA). In this study, samarium nitrate hexahydrate was mixed into artificial diet for boll weevils, Anthonomus grandis grandis Boheman (Coleoptera: Curculionidae), at different dosages and in water used to irrigate cotton, Gossypium hirsutum L. Samarium was detected in adult boll weevils fed on the samarium-labeled diet, but not after 5 or 10 days of being switched to non-labeled diet, even if the insects were given labeled diet for as long as 7 consecutive days. Introduced in irrigation water, 1% samarium (m/m) was detectable in cotton squares and leaf tissue. However, boll weevil adults fed samarium-labeled squares did not retain detectable levels of samarium, nor did boll weevil adults reared to adulthood from samarium-labeled squares. Fourth instar beet armyworms, Spodoptera exigua (Hubner) (Noctuidae: Lepidoptera), fed on samarium-labeled cotton leaves obtained enough samarium for NAA detection, but adult moths reared from them did not have detectable amounts of samarium. Although samarium can be useful as a marker when insects are presented with a continuous pulse of the label, elements that are assimilated by the insect would be more useful if a continuous infusion of the marker cannot be provided. PMID- 16713274 TI - Cross-section measurements for the reactions of 14 MeV neutrons on indium isotopes. AB - The cross sections for the reactions (115)In(n, p)(115g)Cd, (115)In(n, alpha)(112)Ag, (115)In(n, 2n)(114m)In, (113)In(n, 2n)(112m)In, (115)In(n, n')(115m)In, and (113)In(n, n')(113m)In induced by 14 MeV neutrons have been measured by activation relative to the (27)Al(n, alpha)(24)Na. Measurements were carried out by gamma-detection using a coaxial HPGe detector. As samples, natural indium has been used. The fast neutrons were produced by the T(d, n)(4)He reaction. The results obtained are compared with existing data. PMID- 16713275 TI - Analysis of neutron and gamma-ray streaming along the maze of NRCAM thallium production target room. AB - Study of the shield performance of a thallium-203 production target room has been investigated in this work. Neutron and gamma-ray equivalent dose rates at various points of the maze are calculated by simulating the transport of streaming neutrons, and photons using Monte Carlo method. For determination of neutron and gamma-ray source intensities and their energy spectrum, we have applied SRIM 2003 and ALICE91 computer codes to Tl target and its Cu substrate for a 145 microA of 28.5 MeV protons beam. The MCNP/4C code has been applied with neutron source term in mode n p to consider both prompt neutrons and secondary gamma-rays. Then the code is applied for the prompt gamma-rays as the source term. The neutron-flux energy spectrum and equivalent dose rates for neutron and gamma-rays in various positions in the maze have been calculated. It has been found that the deviation between calculated and measured dose values along the maze is less than 20%. PMID- 16713276 TI - Mechanism of the 15N(p,alpha)12C reaction at Ep=9.09-43.7 MeV. AB - The spectroscopic-factor amplitudes for the triton-cluster transfer are used in distorted wave born approximation (DWBA) analyses for the first six (12)C states, up to E(x)=17.76 MeV, in the reaction (15)N(p,alpha) for E(p)=9.09-43.7 MeV. Agreements are obtained for most (12)C states between the theoretically predicted spectroscopic factors (S) and both of experimental and theoretical forward integrated cross-section values (at E(p) approximately 43.7 MeV) and their sequence states with the same nuclear parameters. The theoretical spectroscopic factors failed to predict the transition strengths for the two (12)C states at 7.6542 and 17.76 MeV, while the DWBA predictions failed to reproduce the strengths of the three (12)C states at 7.6542, 14.083 and 17.76 MeV. This is probably, in the first case, due to the inclusion of other direct mechanisms and (or) to the multi-step processes in their mechanism, while for the second case, it is may be due to either the angular momentum mismatching effect or the choice of the optical parameters. The agreement between theoretical and experimental data for the first six investigated (12)C states indicate the success of the Cohen-Kurath wave-functions itself and their accuracy. They also indicate the success of the model of calculations of spectroscopic factors. PMID- 16713277 TI - Theoretical and experimental determination of dosimetric characteristics for ADVANTAGE Pd-103 brachytherapy source. AB - ADVANTAGE Pd-103 brachytherapy source has been recently introduced by IsoAid for prostate permanent implants. Dosimetric characteristics (Dose rate constant, radial dose function, 2D-, and 1D-anisotropy functions) of this source model have been determined using both theoretical and experimental methods, following the updated TG-43U1 protocol. Derivation of the dose rate constant was based on recent NIST WAFAC calibration performed in accordance with the 1999 Standards. Measurements were performed in Solid Water using LiF TLD chips and the theoretical calculations were performed in Solid Water and liquid water phantom materials using PTRAN Monte Carlo code. The results of the Monte Carlo simulation have shown a dose rate constant of 0.69 cGyh(-1) U(-1) in liquid water and 0.67 cGyh(-1) U(-1) in Solid Water medium. The measured dose rate constant in Solid Water was found to be 0.68+/-8% cGyh(-1) U(-1), which is in a good agreement (within +/-5%) to the Monte Carlo simulated data. The 2D- and 1D-anisotropy functions of the ADVANTAGE Pd-103 source were calculated for radial distances ranging from 0.5 to 5.0 cm. Radial dose function was determined for radial distances ranging from 0.2 to 8.0 cm using line source approximation. All these calculations are based on L(eff) equal to 3.61 cm, calculated following TG-43U1 recommendations. The tabulated data for 2D-anisotropy function, 1D-anisotropy function, dose rate constant and radial dose function have been produced for clinical application of this source model. PMID- 16713278 TI - Proteasome dysfunction in aged human alpha-synuclein transgenic mice. AB - A deficit in proteasome function in Parkinson's disease has been speculated. We characterized the ubiquitin-proteasome system in three regions of brain from transgenic and nontransgenic littermates. Mice expressing a doubly mutated form of human alpha-synuclein had significant impairments whereas mice expressing the wild-type gene had lesser changes compared to nontransgenic littermates. Significant abnormalities in line hm2 alpha-SYN-39 included declines in 20S mediated proteolytic activity, the level of the 19S proteasome subunits Rpt1 and Rpn2, and the level of soluble total high MW ubiquitin cross-reacting proteins. Line hw alpha-SYN-5 had significant, but restricted proteasome abnormalities. The severity of impairment was proportional to the substantia nigra dopaminergic neuronal loss previously identified. There were significant correlations between the level of Rpn2 with the level of Rpt1, the activity of the 20S proteasome, and the level of soluble high MW ubiquitin cross-reacting proteins. These abnormalities in symptomatic line hm2 alpha-SYN-39 mice are consistent with abnormalities identified in tissue from patients with Parkinson's disease. PMID- 16713279 TI - Exogenous cytokinin treatment maintains cyclin homeostasis in rice seedlings that show changes of cyclin expression when the photoperiod is rapidly changed. AB - Cyclin is a fundamental regulator of the plant cell cycle. Five different types of cyclin genes (the A-, B-, C-, D-, and H-types) have been reported in Oryza sativa. However, except for Os;cycA1;1, Os;cycB2;1, and Os;cycB2;2, the mechanisms of expression of these cyclin genes have not yet been studied. The interactions of cyclins with cytokinin, an important trigger for cell cycle regulation, have also not been well studied. Here we used semi-quantitative RT PCR in rice seedlings to analyze the effect of cytokinin on photomorphogenesis and the expression of six cyclin genes. Fifteen-day-old seedlings were grown in a 16/8 h light/dark cycle and then transferred to either constant light or constant dark. The expression of all the cyclin genes tested, except the C-type, decreased after 1 hour in the dark, but did not change after transfer to the light or when kinetin was added to the medium. Similarly, seedlings grown in the dark had decreased expression of the cyclin genes, except Os;cycB2;2, after transfer to the light, a decrease that was prevented by kinetin treatment. Thus, exogenous cytokinin plays an important role in maintaining homeostasis of cyclin gene expression following rapid changes of photoperiod. PMID- 16713280 TI - Phenothiazinium-based photobactericidal materials. AB - Photosensitisers based on the phenothiazinium chromophore have been reported to be bactericidal on illumination via a singlet oxygen-mediated mechanism. The current work reports on the immobilisation of the methylene blue analogue, new methylene blue, in polymer resins, and the preliminary development of photobactericidal materials. The resultant materials exhibited measurable levels of singlet oxygen and were photobactericidal against both Staphylococcus epidermidis and Escherichia coli. Such materials may be of use in surface disinfection/infection control in the healthcare milieu. PMID- 16713281 TI - Stretch and CO2 modulate the inflammatory response of alveolar macrophages through independent changes in metabolic activity. AB - Ventilatory-induced strain can exacerbate acute lung injury (ALI). Current ventilation strategies favour low tidal volumes and high end-expiratory volumes to 'rest' the lung, but can lead to an increase in CO2. Alveolar macrophages (AM) play a pivotal role in ALI through the release of inflammatory mediators. The effect of physical strain and CO2 on the release of pro-inflammatory mediators was examined in isolated rat AM. AM were cultured on IgG-coated silastic membranes with or without lipopolysaccharide (LPS) and 5% or 20% CO2 and subjected to a repetitive sinusoidal mechanical strain (30%, 60 cycles/min) for 4 h. Cell viability and metabolic activity were assessed. In both the presence and absence of LPS, physical strain increased metabolic activity by approximately 5%, while 20% CO2 decreased metabolic activity by approximately 40%. Twenty per cent CO2 decreased TNF-alpha secretion by approximately 45%, without affecting cell viability. Physical strain enhanced LPS-induced secretion of TNF-alpha by 1.5%, but not IL-6 or CINC-1. Hence, the effects of both CO2 and physical strain are mediated independently through changes in AM metabolic activity. Physical strain is not a major determinant of TNF-alpha, IL-6 or CINC-1 in AM. Our results confirm that high CO2 can lessen the TNF-alpha inflammatory response of AM. PMID- 16713282 TI - Effect of tumor necrosis factor alpha and infliximab on apoptosis of B lymphocytes infected or not with Epstein-Barr virus. AB - Chronic inflammation and immunosuppressive therapies increase the risk of non Hodgkin's lymphoma associated or not with Epstein-Barr virus (EBV) infection. A possible link between infliximab treatment and increased risk of lymphoma has been suggested. Indeed, infliximab induces apoptosis of monocytes and activated T lymphocytes, but its effect on B lymphocytes infected or not with EBV is unknown. Secreted tumor necrosis factor (TNF) alpha and the expression level of TNF receptor 1 (TNFR1) and TNFR2 were compared in EBV-positive and negative B-cell lines. The impact of TNFalpha and infliximab on apoptosis of EBV-positive cells was analyzed regarding the activity of NF-kappaB. Increased expression of TNFalpha in EBV-positive cells suggested that infliximab could affect their survival. However, TNFalpha or infliximab incubation had no effect on apoptosis of EBV-positive cells. Loss of NF-kappaB activity sensitized lymphoblastoid cell lines to TNFalpha-induced apoptosis, but no direct effect of infliximab on apoptosis was detected. On the basis of our in vitro data, neither TNFalpha nor infliximab has a direct effect on apoptosis of B lymphocytes and EBV-positive cell lines. Thus, if an increased incidence of lymphoma were induced by TNFalpha blockers, it would not involve a direct effect on B cells but rather an impaired immune surveillance by T cells. PMID- 16713283 TI - Oncostatin M induces cell detachment and enhances the metastatic capacity of T 47D human breast carcinoma cells. AB - Oncostatin M (OSM), an IL-6 family cytokine, has previously been shown to increase migration of several breast cancer cell lines in vitro. Our studies report additional effects of OSM treatment on the human breast carcinoma cell line T-47D. OSM treatment alters T-47D cell morphology from a normal epithelial phenotype to a mesenchymal-like phenotype that is associated with cell detachment from substratum. These effects are also seen with H3922 human breast cancer cells. OSM treatment of T-47D cells for 5-8 days leads to a three-fold increase in cell detachment. OSM-induced detachment of T-47D cells is blocked by the protein kinase inhibitors UO126 and bisindolylmaleimide, indicating a role for MAP kinases and protein kinase C in OSM signaling events that regulate cell detachment. T-47D cells induced to detach by OSM have a reduced capacity to re adhere to laminin in comparison to other extracellular matrix components. Detached multi-cell aggregates of T-47D cells are viable, whereas detached single cells appear apoptotic. In addition, OSM treatment induces the secretion of the lysosomal proteases cathepsins D and L from T-47D cells, which have been implicated in invasion and metastasis. Importantly, OSM-treated T-47D cells show a 250% increase in invasive capacity as measured by the Matrigel invasion chamber assay. Collectively, these data demonstrate that OSM induces a motile/invasive phenotype in T-47D cells in vitro, and suggest that OSM may enhance metastasis in vivo. Our results suggest that OSM itself may be a valid therapeutic target. PMID- 16713284 TI - Evolution of cytokine responses: IL-1beta directly affects intracellular Ca2+ concentration of teleost fish leukocytes through a receptor-mediated mechanism. AB - In this work we studied the biological activities of recombinant IL-1beta from the teleosts sea bass (Dicentrarchus labrax) and rainbow trout (Oncorhynchus mykiss) by investigating the effects induced on intracellular Ca2+ concentrations ([Ca2+]i) of spleen leucocytes. Splenocytes were loaded with the Ca2+-permeant Fura-2AM, and then stimulated with rIL-1beta. The emitted fluorescence was read for 5 min at 1 min intervals on a dual excitation fluorescence fluorimeter. Results showed that rIL-1beta induced in both species a rise in [Ca2+]i, and a subsequent decrease until 5 min after stimulation. The stimulating effect was dose-dependent in both species reaching a plateau at 200 ng/ml of rIL-1beta, was abolished by heat-treatment of rIL-1beta, and affected in a dose-dependent fashion by treatment of leucocytes with trypsin. These features suggested a functional IL-1 receptor was involved in the binding. The observed rise in [Ca2+]i was not detected in human PBMC and was species-specific, since rIL-1beta from sea bass, trout, and human were unable to interfere each other in the assay. Moreover, incubation of splenocytes with rIL-1beta induced a rapid tyrosine phosphorylation of a 24 kDa polypeptide in both species. This work represents the first evidence of a direct effect on [Ca2+]i induced by IL-1beta and suggests that in the evolution of IL-1 activities, teleost fishes display a peculiar IL-1 associated behaviour that is lacking in mammals. PMID- 16713285 TI - I(f) in non-pacemaker cells: role and pharmacological implications. AB - Pacemaker channels play a major role in the generation of sinoatrial rhythmic activity. However, their expression is not confined to specialized myocardial cells, such as primary and subsidiary pacemakers. Electrophysiological and molecular data collected over the last ten years have demonstrated that f channels are also present in non-pacemaker cardiomyocytes, and become upregulated in cardiac hypertrophy and failure. Mislocalized expression and/or overexpression of f-channels are a consequence of electrophysiological remodeling and, from a clinical point of view, may represent an arrhythmogenic mechanism in heart failure, a condition associated with a high risk for sudden cardiac death. The potential arrhythmogenic role of I(f) and the availability of selective f-channel blockers cause I(f) to be a suitable therapeutic target in heart disease. PMID- 16713286 TI - Influence of the matrix on analyte fragmentation in atmospheric pressure MALDI. AB - In this paper, we report the measurement of the degree of analyte fragmentation in AP-MALDI as a function of the matrix and of the laser fluence. The analytes include p-OCH3-benzylpyridinium, three peptides containing the sequence EEPP (which cleave very efficiently at the E-P site), and three deoxynucleosides (dA, dG, and dC), which lose the neutral sugar to give the protonated base. We found that the matrix hardness/softness was consistent when comparing the analytes, with a consensus ranking from hardest to softest: CHCA >> DHB > SA approximately THAP > ATT > HPA. However, the exact ranking can be fluence-dependent, for example between ATT and HPA. Our goal here was to provide the scientific community with a detailed dataset that can be used to compare with theoretical predictions. We tried to correlate the consensus ranking with different matrix properties: sublimation or decomposition temperature (determined using thermogravimetry), analyte initial velocity, and matrix proton affinity. The best correlation was found with the matrix proton affinity. PMID- 16713288 TI - Mass spectrometric separation and quantitation of overlapping isotopologues. H2O/HOD/D2O and H2Se/HDSe/D2Se mixtures. AB - Three conceptually different mathematical methods are presented for accurate mass spectrometric determination of H2O/HOD/D2O and H2Se/HDSe/D2Se concentrations from mixtures. These are alternating least-squares, weighted two-band target entropy minimization, and a statistical mass balance model. The otherwise nonmeasurable mass spectra of partially deuterated isotopologues (HOD and HDSe) are mathematically constructed. Any recorded isotopologue mixture mass spectra are then deconvoluted by least-squares into their components. This approach is used to study the H2O/D2O exchange reaction, and is externally validated gravimetrically. The H2O/D2O exchange equilibrium constant is also measured from the deconvoluted 70 eV electron impact GC/MS data (K = 3.85 +/- 0.03). PMID- 16713289 TI - A simple method for estimating activation energies using the fragmentation yield: collision-induced dissociation of iron(II)-phenanthroline complexes in an electrospray ionization mass spectrometer. AB - The gas-phase stabilities of Fe(Phi)3(2+) complexes, where Phi represents the 1,10-phenanthroline, 5-chloro-1,10-phenanthroline, 5-methyl-1,10-phenanthroline, 3,4,7,8-tetramethyl-1,10-phenanthroline, and 4,7-diphenyl-1,10-phenanthroline ligands were investigated by collision-induced dissociation (CID) in the capillary-first skimmer region upon changing the voltage difference between the capillary and the skimmer. The loss of only one ligand from the Fe(Phi)3(2+) complexes was observed with each of the phenanthroline ligands studied. An increase in the voltage difference between the capillary and the skimmer resulted in a higher fragmentation yield as calculated from the intensity of the precursor and the fragment ion. The fragmentation yield versus capillary-skimmer voltage difference plots were evaluated by means of the Arrhenius and the Rice-Ramsperger Kassel (RRK) model by fitting the model parameters to the experimental data. Both models yielded practically the same results. In addition, if the internal energy gained through the capillary-skimmer region is estimated correctly, the approximate value of the critical energy (activation energy) for fragmentation can be extracted from the fragmentation yield versus capillary-skimmer voltage difference plots. It was found that the gas-phase stabilities of the Fe(Phi)3(2+) complexes are nearly identical except for the more stable Fe(II)-4,7-diphenyl 1,10-phenanthroline complex. The critical energy for fragmentation was estimated to be approximately 1.2 and 0.9 eV for the Fe(II)- 4,7-diphenyl-1,10 phenanthroline, and the other complexes, respectively. PMID- 16713287 TI - Electrospray mass spectrometry of NeuAc oligomers associated with the C fragment of the tetanus toxin. AB - The Clostridial neurotoxins, botulinum and tetanus, gain entry into motor neurons by binding to the sialic or N-acetylneuraminic acid (NeuAc) residues of gangliosides and specific protein receptors attached to the cell's surface. While the C-fragment of tetanus toxin (TetC) has been identified to be the ganglioside binding domain, remarkably little is known about how this domain discriminates between the structural features of different gangliosides. We have used electrospray ionization mass spectrometry (ESI-MS) to examine the formation of complexes between TetC and carbohydrates containing NeuAc groups to determine how NeuAc residues contribute to ganglioside binding. ESI-MS was used to obtain an estimate of the dissociation constants (KD values) for TetC binding to a number of related NeuAc-containing carbohydrates (sialyllactose and disialyllactose), as well as six (NeuAc)n oligomers (n = 1-6). KD values were found to range between approximately 10-35 microM. The strength of the interactions between the C fragment and (NeuAc)n are consistent with the topography of the targeting domain of tetanus toxin and the nature of its carbohydrate binding sites. These results suggest that the targeting domain of tetanus toxin contains two binding sites that can accommodate NeuAc (or a dimer) and that NeuAc may play an important role in ganglioside binding and molecular recognition, a process critical for normal cell function and one frequently exploited by toxins, bacteria, and viruses to facilitate their entrance into cells. PMID- 16713290 TI - Carbon nanotubes (2,5-dihydroxybenzoyl hydrazine) derivative as pH adjustable enriching reagent and matrix for MALDI analysis of trace peptides. AB - A functionalized carbon nanotube (CNT), CNT 2,5-dihydroxybenzoyl hydrazine derivative, was synthesized and used as both pH adjustable enriching reagent and matrix in matrix assisted laser desorption/ionization mass spectrometry (MALDI MS) analysis of trace peptides. The derivative reagent, 2,5-dihydroxybenzoyl hydrazine, introduced phenolic hydroxyl and phenyl groups to the surface of the CNT. The former group can provide adjustable surface charge and a source of protons for chemical ionization, and the latter helps to keep strong ultraviolet absorption for enhancing pulsed laser desorption and ionization. It was found that the functionalized CNT was less twisted in a basic condition (pH 10.5), which afforded an increased surface area to volume ratio for adsorption towards trace peptides. However, functionalized CNT becomes deposited in an acidic condition (pH 5) and can be isolated readily from the sample solutions once the nanoparticles have trapped the target analytes, thus providing a novel and convenient alternative method for quick isolation. Compared with the previously reported method on enriching analytes using the pristine CNT, it is observed that the detection limit for analytes can be greatly improved due to enhancing adsorption capacity of the functionalized CNT. Moreover, peptide mixture at concentration as low as 0.01 pg/microL still can be detected after enrichment mediated by the functionalized CNT, while it is difficult to be detected without enrichment at concentration 0.1 pg/microL using alpha-cyano-4-hydroxycinnamic acid (CHCA) as matrix. Therefore, high efficiency of adsorption and enrichment towards trace peptides can be achieved by adjusting pH value of the functionalized CNT dispersion. PMID- 16713292 TI - Distonoid ions. AB - By Yates, Bouma, and Radom's definition, distonic radical ions are those formally arising by ionization of diradicals or zwitterionic molecules (including ylides). These ions differ, therefore, from conventional radical ions by displaying the charge site and unpaired electron site (spin) localized mandatorily on separate atoms or group of atoms; that is, these sites are separated in all of their major resonance forms. Many conventional radical ions with a major resonance form in which charge and spin sites reside formally on the same atom or group of atoms display, however, high degree of discretionary (non-mandatory) charge-spin separation. By analogy with the metal/metalloid terminology, we propose that these distonic-like radical ions be classified as distonoid ions. Radical ions would, therefore, be divided into three sub-classes: conventional, distonic, and distonoid ions. B3LYP/6-311 + G(d,p) calculations for a proof-of-principle set of radical cations are used to demonstrate the existence of many types of distonoid ions with a high degree of discretionary charge-spin separation. Reliable calculations are indispensable for probing distonoid ions, since an ion that was expected to be distonoid (by the analysis of its resonance forms) is shown by the calculations to display a characteristic conventional-ion electronic distribution. Similarly to many distonic radical ions, and in sharp contrast to a conventional radical ion (ionized 1,4-dioxane), the gas-phase intrinsic bimolecular reactivity with selective neutrals of a representative distonoid ion, ionized 2-methylene 1,3-dioxolane, is found to include dual ion-radical type reactions. PMID- 16713291 TI - Analysis of protein-protein interaction surfaces using a combination of efficient lysine acetylation and nanoLC-MALDI-MS/MS applied to the E9:Im9 bacteriotoxin- immunity protein complex. AB - To understand how proteins perform their function, knowledge about their structure and dynamics is essential. Here we use a combination of an efficient chemical lysine acetylation reaction and nanoLC-MALDI tandem mass spectrometry to probe the accessibility of every lysine residue in a protein complex. To demonstrate the applicability of this approach, we studied the interaction between the DNase domain of Colicin E9 (E9) and its immunity protein Im9. Free E9 and E9 in complex with Im9 were rapidly acetylated, followed by proteolytic digestion and analysis by LC-MALDI-TOF/TOF MS/MS. Acetylated peptides could be filtered out of the complex peptide mixtures using selective ion chromatograms of the specific immonium marker ions. Additionally, isobaric acetylated peptides, acetylated at different sites, could be separated by their LC retention times. The combination of LC and MALDI-TOF/TOF MS/MS provided information about the amount of acetylation on each individual lysine even for peptides containing several lysine residues. In general, our data agree well with those derived from the crystal structure of E9 and the E9:Im9 complex. Interestingly, next to in the binding interface expected lysines, K89 and K97, two from the crystal structure data unexpected lysines, K81 and K76, were observed to become less exposed upon Im9 binding. Moreover, K55 and K63, positioned in the predicted DNA binding region, were also found to be less accessible upon Im9 binding. These findings may illustrate some of the described differences in the solution-phase structure of the E9:Im9 complex compared with the crystal structure. PMID- 16713293 TI - The transcription factor ATF-3 promotes neurite outgrowth. AB - Dorsal root ganglion (DRG) neurons regenerate after a peripheral nerve injury but not after injury to their axons in the spinal cord. A key question is which transcription factors drive the changes in gene expression that increase the intrinsic growth state of peripherally injured sensory neurons? A prime candidate is activating transcription factor-3 (ATF-3), a transcription factor that we find is induced in all DRG neurons after peripheral, but not central axonal injury. Moreover, we show in adult DRG neurons that a preconditioning peripheral, but not central axonal injury, increases their growth, correlating closely with the pattern of ATF-3 induction. Using viral vectors, we delivered ATF-3 to cultured adult DRG neurons and find that ATF-3 enhances neurite outgrowth. Furthermore, ATF-3 promotes long sparsely branched neurites. ATF-3 overexpression did not increase c-Jun expression. ATF-3 may contribute, therefore, to neurite outgrowth by orchestrating the gene expression responses in injured neurons. PMID- 16713295 TI - Evidence for a dimeric assembly of two titin/telethonin complexes induced by the telethonin C-terminus. AB - The Z-disk region defines the lateral boundary of the sarcomere and requires a high level of mechanical strength to provide a stable framework for large filamentous muscle proteins. The level of complexity at this area is reflected by a large number of protein-protein interactions. Recently, we unraveled how the N terminus of the longest filament component, the giant muscle protein titin, is assembled into an antiparallel (2:1) sandwich complex by the N-terminal titin binding segment of the Z-disk ligand telethonin/T-cap [Zou, P., Pinotsis, N., Lange, S., Song, Y.H., Popov, A., Mavridis, I., Mayans, O.M., Gautel, M., Wilmanns, M., 2006. Palindromic assembly of the giant muscle protein titin in the sarcomeric Z-disk. Nature 439, 229-233]. In this contribution, we present structural data of a related complex of the titin N-terminus with full-length telethonin. The C-terminus of telethonin remains invisible, suggesting that it does not fold into a defined structure even in the presence of titin. In contrast to the structure with truncated telethonin, a dimer of two titin/telethonin complexes is formed within the crystal environment, potentially indicating the formation of higher oligomers. We further investigated the structure and dynamics of this assembly by small-angle X-ray scattering, circular dichroism, and in vivo complementation data. The data consistently indicate the involvement of the C terminal part of telethonin into the assembly of two titin/telethonin complexes. PMID- 16713294 TI - Site-specific 3D imaging of cells and tissues with a dual beam microscope. AB - Current approaches to 3D imaging at subcellular resolution using confocal microscopy and electron tomography, while powerful, are limited to relatively thin and transparent specimens. Here we report on the use of a new generation of dual beam electron microscopes capable of site-specific imaging of the interior of cellular and tissue specimens at spatial resolutions about an order of magnitude better than those currently achieved with optical microscopy. The principle of imaging is based on using a focused ion beam to create a cut at a designated site in the specimen, followed by viewing the newly generated surface with a scanning electron beam. Iteration of these two steps several times thus results in the generation of a series of surface maps of the specimen at regularly spaced intervals, which can be converted into a three-dimensional map of the specimen. We have explored the potential of this sequential "slice-and view" strategy for site-specific 3D imaging of frozen yeast cells and tumor tissue, and establish that this approach can identify the locations of intracellular features such as the 100 nm-wide yeast nuclear pore complex. We also show that 200 nm thick sections can be generated in situ by "milling" of resin-embedded specimens using the ion beam, providing a valuable alternative to manual sectioning of cells and tissues using an ultramicrotome. Our results demonstrate that dual beam imaging is a powerful new tool for cellular and subcellular imaging in 3D for both basic biomedical and clinical applications. PMID- 16713296 TI - Mechanical manipulation of Alzheimer's amyloid beta1-42 fibrils. AB - The 39- to 42-residue-long amyloid beta-peptide (Abeta-peptide) forms filamentous structures in the neuritic plaques found in the neuropil of Alzheimer's disease patients. The assembly and deposition of Abeta-fibrils is one of the most important factors in the pathogenesis of this neurodegenerative disease. Although the structural analysis of amyloid fibrils is difficult, single-molecule methods may provide unique insights into their characteristics. In the present work, we explored the nanomechanical properties of amyloid fibrils formed from the full length, most neurotoxic Abeta1-42 peptide, by manipulating individual fibrils with an atomic force microscope. We show that Abeta-subunit sheets can be mechanically unzipped from the fibril surface with constant forces in a reversible transition. The fundamental unzipping force (approximately 23 pN) was significantly lower than that observed earlier for fibrils formed from the Abeta1 40 peptide (approximately 33 pN), suggesting that the presence of the two extra residues (Ile and Ala) at the peptide's C-terminus result in a mechanical destabilization of the fibril. Deviations from the constant force transition may arise as a result of geometrical constraints within the fibril caused by its left handed helical structure. The nanomechanical fingerprint of the Abeta1-42 is further influenced by the structural dynamics of intrafibrillar interactions. PMID- 16713297 TI - Application of a new vector system for efficient protein purification in the crystallization of PA5104/ORF from Pseudomonas aeruginosa. AB - We have developed a new T7-based vector system for rapid purification and high throughput capability applicable for structural studies. The system allows purification of target proteins to homogeneity in two steps with a single Ni affinity column. The first step relies on affinity purification of the N-terminal His-tagged protein in the conventional way, eluting the protein with imidazole. Addition of a His-tagged 3C protease to cleave the His-tag permits a second pass through the nickel column, this time all impurities bind to the column while the pure protein does not. This has the major advantage of quickly removing the residual contaminating proteins that are associated with nickel affinity purification as well as the protease and His-tag. Here, we describe the application of this system to over-express and purify ORF PA5104 from Pseudomonas aeruginosa. The protein was successfully crystallized and crystals were shown to diffract to atomic resolution. Additionally preliminary X-ray diffraction analysis of two crystals forms is presented, one diffracting to 1.9 A and the other to 0.96 A resolution. PMID- 16713298 TI - Solubility properties and specific assembly pathways of the B-type lamin from Caenorhabditis elegans. AB - Lamins are nucleus-specific intermediate filament (IF) proteins that together with a complex set of membrane proteins form a filamentous meshwork tightly adhering to the inner nuclear membrane and being associated with the nuclear pore complexes. This so-called nuclear lamina provides mechanical stability and, in addition, has been implicated in the spatial organization of the heterochromatin. While increasing knowledge on the biological function of lamins has been obtained in recent years, the assembly mechanism of lamin filaments at the molecular level has remained largely elusive. Therefore, we have now more systematically investigated lamin assembly in vitro. Using Caenorhabditis elegans lamin, which has been reported to assemble into 10-nm filaments under low ionic strength conditions, we investigated the assembly kinetics of this protein into filaments in more detail using both His-tagged and un-tagged recombinant proteins. In particular, we have characterized distinct intermediates in the filament assembly process by analytical ultracentrifugation, electron and atomic force microscopy. In contrast to the general view that lamins assemble only slowly into filaments, we show that in vitro association reactions are extremely fast, and depending on the ionic conditions employed, significant filamentous assemblies form within seconds. PMID- 16713299 TI - Hendecad repeat in segment 2A and linker L2 of intermediate filament chains implies the possibility of a right-handed coiled-coil structure. AB - The conformation adopted by intermediate filament chains (IF) has been described in terms of a central rod domain with four, alpha-helical, left-handed coiled coil segments (1A, 1B, 2A, and 2B) joined by linkers (L1, L12, and L2, respectively). The rod domain is terminated at its N- and C-terminal ends by "globular" head and tail domains, respectively. This analysis, initially undertaken about 20-25 years ago, was based on the recognition of an underlying heptad substructure in the sequence of the rod domain, the presence of which can be directly associated with an alpha-helical coiled-coil structure. In this work, a hendecad sequence motif that is closely related to the heptad repeat but which is nonetheless significantly different from it has been recognized in the primary structure of segments 2A and linker L2. This motif, which is 11 residues long and structurally equivalent to a true heptad plus another heptad with an inclusive stutter, is consistent with the chains adopting a continuous right-handed coiled coil structure with a long-period pitch length. It is therefore predicted that segment 2 as a whole may have a coiled-coil conformation with both right-handed (2A+L2) and left-handed (2B) regions. The changeover in handedness would be expected to occur at the C-terminal end of linker L2 and N-terminal end of segment 2B. PMID- 16713301 TI - Human hair keratin-associated proteins: sequence regularities and structural implications. AB - In this paper, we undertake a sequence analysis of the human keratin-associated proteins (KAP). This analysis has revealed two fundamental pentapeptide quasi repeats (A and B) of the form C-C-X-P-X and C-C-X-S/T-S/T, respectively. The A repeats are also commonly found in two subforms A1 and A2, -C-C-Q-P-X and C-C-R-P X, respectively-similar to those found in sheep wool 30-40 years previously. Some high-sulphur and ultra-high sulphur proteins contain predominantly A repeats or B repeats but not regular combinations of them, whereas others are characterised by a contiguous pair of pentapeptide repeats that largely (though imperfectly) alternate to generate decapeptide motifs of the form AB, A1B or A2B. The A and B repeats sometimes occur in complex runs and can generate both 19- and 20-residue repeats of the form BABB' or BA1AA, respectively, where the prime indicates a motif truncated by one residue. Likewise, a 42-residue repeat with BA1BXAAAB (40 residues) separated by a di-serine (two residues) has been observed in an ultra high sulphur protein from cuticle. To understand the possible conformations adopted by the A and B motifs, a search was initiated of the PDB structural database for a number of overlapping pentapeptide repeats. The total number of matches was 658 and these were found in 451 different proteins. From representative and unique structures the means and standard deviations were calculated for the Phi(i) and Psi(i) angles for the C-C-X-P-X and the C-C-X-S/T S/T motifs. Molecular modelling has been employed to represent the "average" structure found from crystallographic and nmr data determined for each motif in other proteins. The conformation of consecutive A repeats with proline residues in the cis state is akin to a string of disulphide bond-stabilised pentapeptide knots between which there is relative freedom of rotation about the single bonds that link them. For B pentapeptides, however, the likelihood that a similar disulphide bond is formed appears much lower. This may give additional conformational flexibility to the chain and hence allow the A pentapeptides greater opportunity to interact appropriately with the IF via disulphide bonds, ionic interactions and/or hydrogen bonding. PMID- 16713302 TI - Structural correlation between collagen VI microfibrils and collagen VI banded aggregates. AB - Collagen VI is a component of the extracellular matrix that is able to form structural links with cells. Collagen VI monomers cross-link into tetramers that come together to form long molecular chains known as microfibrils. Collagen VI tetramers are also the most likely candidates for the formation of banded aggregates with an axial periodicity of about 105 nm that are seen in the retinas of people suffering from age-related macular degeneration and Sorsby's fundus dystrophy, in the vitreous of patients with full thickness macular holes and in the intervertebral discs of normal individuals. Here, a protocol is developed to carry out a structural comparison between the microfibrils, which are known to be made of collagen VI tetramers, and the banded aggregates. The comparison shows that the banded aggregates are easily explained as being a lateral assembly of microfibrils, thus supporting the hypothesis that they too are made of collagen VI. Understanding the role played by the collagen VI aggregates in normal and pathological conditions will help to throw light on the pathologies with which they are associated. PMID- 16713303 TI - Effect of sulfide on the immune response and susceptibility to Vibrio alginolyticus in the kuruma shrimp Marsupenaeus japonicus. AB - Kuruma shrimp Marsupenaeus japonicus held in 34 per thousand seawater were injected with tryptic soy broth (TSB)-grown Vibrio alginolyticus (2.7x10(6)cfu shrimp(-1)), and then placed in water containing concentrations of sulfide at 0 (control), 51, 106, 528 and 1050microgl(-1), respectively. After 12-144h, mortality of V. alginolyticus-injected shrimp exposed to 528 and 1102microgl(-1) sulfide was significantly higher than that of shrimp exposed to 51microgl(-1) sulfide and the control solution. In another experiment, M. japonicus which had been exposed to 0, 56, 112, 525 and 1076microgl(-1) sulfide for 6, 12, 24 and 48h were examined for immune parameters, and phagocytic activity and clearance efficiency of V. alginolyticus. Sulfide concentrations at 525microgl(-1) or greater for 12h resulted in decreased total haemocyte count (THC) and phenoloxidase activity, phagocytic activity and bacterial clearance efficiency, whereas a sulfide concentration at 1076microgl(-1) for 24h caused a significant increase in respiratory burst and superoxide dismutase activity of M. japonicus. It is concluded that concentrations of sulfide at 528microgl(-1) or greater increased the susceptibility of M. japonicus against V. alginolyticus infection by a depression in immune ability. The increased production of superoxide anion by M. japonicus exposed to 525microgl(-1) sulfide or greater was considered to be cytotoxic to the host. PMID- 16713304 TI - Infectious pancreatic necrosis virus suppresses type I interferon signalling in rainbow trout gonad cell line but not in Atlantic salmon macrophages. AB - RTG-P1 cells are a rainbow trout fibroblastic cell line permanently transfected with the luciferase gene under the control of the Mx promoter. On exposure to interferon (IFN) or IFN inducing agents, the cells produce luciferase. IPNV did not induce luciferase production up to 24h post-infection but did not suppress constitutive luciferase production. Furthermore, IPNV suppressed luciferase production induced by poly I:C. RT-PCR analysis of IPNV infected cells showed IFN gene transcription from 6h post-infection with increasing expression up to 24h. Housekeeping genes beta-actin and GAPDH were also expressed along with upregulation of IRF1 and slight upregulation of STAT1. When RTG-P1 cells were stimulated with IFN, Mx transcripts, measured by qRT-PCR, peaked at 3-6h and thereafter fell to low levels, but in the presence of IPNV, Mx transcription at this time was significantly suppressed but continued to rise gradually. Luciferase production was lower in infected cells at 12h post-infection but not significantly after 24h. These results indicate that, in non-stimulated RTG-P1 cells, while IPNV induces IFN transcription, activation of Mx expression is suppressed. Furthermore, when stimulated by IFN, the rate of Mx transcription is significantly suppressed by the virus. This would probably give time for the virus to replicate rapidly in the early phases of infection. Contrary to the fibroblastic cell line, IPNV stimulated IFN production by salmon macrophages in vitro at least as strongly as poly I:C, with no suppression of the IFN response to poly I:C, and the virus persisted for up to 9 days without causing CPE. PMID- 16713305 TI - Measurements of tissue T1 spin-lattice relaxation time and discrimination of large draining veins using transient EPI data sets in BOLD-weighted fMRI acquisitions. AB - The signal intensity during the dynamic approach to the equilibrium state of longitudinal magnetization is a function of sequence parameters, such as repetition time and flip angle, and depends on tissue characteristics, including longitudinal relaxation time of stationary tissue and the rate of blood inflow. A method is presented to extract information from data acquired during the transient state prior to T1 equilibrium using echo-planar acquisitions in T2* weighted functional magnetic resonance imaging (fMRI) experiments. A voxel in a single slice acquisition is assumed to contain either stationary tissue or large vessels with flowing blood. Models are presented to characterize longitudinal magnetization relaxation of heterogeneous stationary tissue and blood inflow. The data were fitted to theoretical models for longitudinal relaxation of stationary tissue and inflowing blood assuming no residual signal prior to each RF excitation. Parameters were estimated at 3 T for each model using least squares estimation. A goodness-of-fit criterion was applied to exclude voxels that have transient data that does not fit the selected (best fit) model. Voxels that best fit the inflow model, measured at various TR and flip angles, were assumed to contain large draining veins and were excluded from functional maps. Histogram analysis of T1 distributions for activated voxels in a visual paradigm demonstrated the distributions are centered at T1 values of gray matter with tails at both sides of the center due to partial voluming of gray matter with white matter and CSF respectively. The mean gray matter volume fraction in activated voxels was about 0.9. The results indicate that transient data sets can provide additional information that is useful for both localization and characterization of the functionally relevant BOLD response. PMID- 16713306 TI - Gender differences in regional cerebral activity during the perception of emotion: a functional MRI study. AB - Whether men activate different brain regions during various emotions compared to women or whether gender differences exist in transient emotional states has been the subject of only few studies. We used event-related functional magnetic resonance imaging (fMRI) to investigate gender differences during the perception of positive or negative emotions. The experiment comprised two emotional conditions (pleasant/unpleasant visual stimuli) during which fMRI data were acquired. Altogether, 38 healthy volunteers (19 males, 19 females) were investigated. When subtracting the activation values of men from those of women, suprathreshold positive signal changes were detected in the right posterior cingulate, the left putamen and the left cerebellum during positive mood induction, and in bilateral superior temporal gyri and cerebellar vermis during negative mood induction. The subtraction of activation values of women from those of men yielded no significant differences. Our findings suggest gender-related neural responses to emotional stimuli and could contribute to the understanding of mechanisms underlying gender-related vulnerability of the prevalence and severity of neuropsychiatric disorders. PMID- 16713307 TI - Phylogeny of the cricket subfamily Eneopterinae (Orthoptera, Grylloidea, Eneopteridae) based on four molecular loci and morphology. AB - The phylogenetic relationships of 39 species of Eneopterinae crickets are reconstructed using four molecular markers (16S rRNA, 12S rRNA, cytochrome b, 18S rRNA) and a large morphological data set. Phylogenetic analysis via direct optimisation of DNA sequence data using parsimony as optimality criterion is done for six combinations of weighting parameter sets in a sensitivity analysis. The results are discussed in a twofold purpose: first, in term of significance of the molecular markers for phylogeny reconstruction in Ensifera, as our study represents the first molecular phylogeny performed for this insect suborder at this level of diversity; second, in term of corroboration of a previous phylogeny of Eneopterinae, built on morphological data alone. The four molecular markers all convey phylogenetic signal, although variously distributed on the tree. The monophyly of the subfamily, that of three over five tribes, and of 10 over 13 genera, are recovered. Finally, previous hypotheses on the evolution of acoustic devices and signals in the Eneopterinae clade are briefly tested, and supported, by our new data set. PMID- 16713308 TI - A simple in vitro model to study the stability of acylglucuronides. AB - INTRODUCTION: Compounds containing the carboxylic functional group (e.g. non steroidal anti-inflammatory drugs) can be metabolized to form acylglucuronides. Acylglucuronides are intrinsically reactive metabolites capable of undergoing hydrolysis, intra-molecular rearrangement, and formation of covalent adducts with proteins, which may generate potential toxicity. The purpose of this study is to develop an in vitro screening model to assess degradation kinetics of acylglucuronides. METHOD: Zomepirac, ibuprofen, gemfibrozil, and compounds A, B, C, and D were incubated in the presence of rat microsomal protein and uridine 5' diphosphoglucuronic acid (UDPGA), followed by addition of human plasma to evaluate degradation kinetics of the acylglucuronides. As a comparison, authentic acylglucuronide standards of zomepirac, ibuprofen, gemfibrozil, and compounds A, B, C, and D were chemically synthesized and were evaluated for degradation kinetics. RESULTS: The results demonstrate that degradation half-life values of acylglucuronides of zomepirac, ibuprofen, gemfibrozil, and compounds A, B, C, and D determined by the in vitro formation/degradation model were in the same rank order with those of the authentic acylglucuronide standards. DISCUSSION: For the seven compounds tested, the model placed the stability of the acylglucuronides formed in vitro in a rank-order consistent with authentic acylglucuronide standards. The method allows for a rapid assessment of the stability of acylglucuronides. PMID- 16713309 TI - Postoperative fever after hemispherectomy: the role of non-infectious factors. AB - Postoperative fever is a usual source of concern among caregivers and patients' family given that it may reflect a wide range of complications. The objective of this paper was to outline the expected postoperative temperature variation after hemispherectomies, and to establish factors that affect this curve. From 1987 to 2003, 30 patients were hemispherectomized in our institution. Among them, 24 patients without clinical diagnosis of infection were selected for this study. Postoperative axillary temperature from each patient was recorded over 10 days. Data was initially analyzed for all 24 patients and, later, for subgroups by (1) surgical technique (functional hemispherectomy, anatomical hemispherectomy, and hemispherotomy) and (2) hemispheric volume of the causative pathology (groups with large, moderately atrophic, and severely atrophic hemispheres). There was a trend of temperature elevation (p=0.06) over the first 4 days in all patients where 14 subjects recorded values over 38.5 degrees C, and 3 over 39.0 degrees C. Temperature curves from subgroups undergoing different techniques presented no statistical differences. However, the group including causative pathologies with larger hemispheres had a higher temperature curve when compared to patients with severely atrophic hemispheres (p<0.05). Our findings support the view that fever is an predictable event after hemispherectomies and causative pathologies with larger hemispheres present higher postoperative temperatures. PMID- 16713310 TI - The relationship between specific tissue lesions and pain severity in persons with knee osteoarthritis. AB - INTRODUCTION: Pain is the most common symptom in knee osteoarthritis (OA), a leading cause of chronic disability, and a major source of the disability attributable to OA in general. Pain severity in knee OA is variable, ranging from barely perceptible to immobilizing. The knee lesions that contribute to pain severity have received little attention. OBJECTIVE: To examine whether worse pathology of specific knee tissues - i.e. cartilage, bone (attrition, cysts, bone marrow lesions, and osteophytes), menisci (tears and subluxation), ligaments, and synovium (synovitis/effusion) - is associated with more severe knee pain. METHODS: One hundred and forty-three individuals were recruited from the community with primary (idiopathic) knee OA, with definite tibiofemoral osteophytes in at least one knee, and at least some difficulty with knee requiring activity. Knee magnetic resonance (MR) images were acquired using coronal T1-weighted spin-echo (SE), sagittal fat-suppressed dual-echo turbo SE, and axial and coronal fat-suppressed, T1-weighted 3D-fast low angle shot (FLASH) sequences. The whole-organ magnetic resonance imaging (MRI) scoring (WORMS) method was used to score knee tissue status. Since summing tissue scores across the entire joint, including regions free of disease, may dilute the ability to detect a true relationship between that tissue and pain severity, we used the score from the worst compartment (i.e. with the poorest cartilage morphology) as our primary approach. Knee pain severity was measured using knee-specific, 100 mm visual analogue scales. In analyses to evaluate the relationship between knee pain severity and lesion score, median quantile regression was used, adjusting for age and body mass index (BMI), in which a 95% CI excluding 0 is significant. RESULTS: The increase in median pain from median quantile regression, adjusting for age and BMI, was significant for bone attrition (1.91, 95% confidence interval (CI) 0.68, 3.13), bone marrow lesions (3.72, 95% CI 1.76, 5.68), meniscal tears (1.99, 95% CI 0.60, 3.38), and grade 2 or 3 synovitis/effusion vs grade 0 (9.82, 95% CI 0.38, 19.27). The relationship with pain severity was of borderline significance for osteophytes and cartilage morphology and was not significant for bone cysts or meniscal subluxation. Ligament tears were too infrequent for meaningful analysis. When compared to the pain severity in knees with high scores for both bone attrition and bone marrow lesions (median pain severity 40 mm), knees with high attrition alone (30 mm) were not significantly different, but knees with high bone marrow lesion without high attrition scores (15 mm) were significantly less painful. CONCLUSION: In persons with knee OA, knee pain severity was associated with subarticular bone attrition, bone marrow lesions, synovitis/effusion, and meniscal tears. The contribution of bone marrow lesions to pain severity appeared to require the presence of bone attrition. PMID- 16713311 TI - Leflunomide--an immunomodulator--induces regression of endometrial explants in a rat model of endometriosis. AB - OBJECTIVE: To test if leflunomide, an immunomodulator, could impede the growth of an ectopic uterine tissue. METHODS: Endometriosis was surgically induced in 26 rats by transplanting an autologous fragment of endometrial tissue onto the inner surface of the abdominal wall. Four weeks later two rats were killed. The volume and weight of the implants were measured. The remaining rats were randomly grouped, and in group 1 no medication was given. To the rats in group 2, 35 mg/kg/d of leflunomide was administered orally. Four weeks later, rats were killed and ectopic uterine tissues were reevaluated morphologically and histologically. A scoring system was used to evaluate preservation of epithelia. RESULTS: Two rats in the control group died 5 weeks after surgery. There was a significant difference in post-treatment spherical volumes (139.1 +/- 92.8 versus 33.5 +/- 12.5 mm3) and explant weights (156.3 +/- 105.6 versus 38.6 +/- 12.6 mg) between the control and leflunomide-treated groups. The epithelia were found to be preserved significantly better in the control group when compared with the leflunomide-treated group (median 2.5 [interquartile range, 1.25] versus median 1.00 [interquartile range, 1.5]). CONCLUSION: Leflunomide appeared to cause regression of experimental endometriosis in rats. PMID- 16713312 TI - Insulin-like growth factor-I regulates vascular endothelial growth factor secretion by human oviductal epithelial cells and stromal fibroblasts. AB - OBJECTIVE: The aim of the current study is to evaluate the effect of insulin-like growth factor-I (IGF-I) on the production of vascular endothelial growth factor (VEGF) in the human fallopian tube. METHODS: Human oviductal epithelial cells (OEC) and oviductal stromal fibroblasts (OSF) were isolated from the ampullary segment of the fallopian tubes of six premenopausal patients in the proliferative phase of the menstrual cycle. The secretion of VEGF165 by cultured OEC and OSF in response to IGF-I was measured using an enzyme-linked immunosorbent assay (ELISA). RESULTS: The secretion of VEGF165 was detected in cultured OEC and OSF under untreated conditions. The secretion of VEGF165 was significantly stimulated with IGF-I administration in these cells. CONCLUSION: The present findings suggest that IGF-I in the local environment may stimulate oviductal vascular permeability by inducing the production of VEGF by oviductal cells through autocrine and paracrine mechanisms. The modulation of the VEGF production in the fallopian tube may contribute to the normal and pathologic processes of oviductal fluid secretion by regulating oviductal vascular permeability during the menstrual cycle and in the peri-implantation period. PMID- 16713314 TI - Transcriptional and bioinformatic analysis of the 56.8 kb DNA region amplified in tandem repeats containing the penicillin gene cluster in Penicillium chrysogenum. AB - High penicillin-producing strains of Penicillium chrysogenum contain 6-14 copies of the three clustered structural biosynthetic genes, pcbAB, pcbC, and penDE [Barredo, J.L., Diez, B., Alvarez, E., Martin, J.F., 1989. Large amplification of a 35-kb DNA fragment carrying two penicillin biosynthetic genes in high penicillin producing strains of Penicillium chrysogenum. Curr. Genet. 16, 453 459; Smith, D.J., Bull, J.H., Edwards, J., Turner, G., 1989. Amplification of the isopenicillin N synthetase gene in a strain of Penicillium chrysogenum producing high levels of penicillin. Mol. Gen. Genet. 216, 492-497.] . The cluster is located in a 56.8 kb DNA region bounded by a conserved TGTAAA/T hexanucleotide that undergoes amplification in tandem repeats [Fierro, F., Barredo, J.L., Diez, B., Gutierrez, S., Fernandez, F.J., Martin, J.F., 1995. The penicillin gene cluster is amplified in tandem repeats linked by conserved hexanucleotide sequences. Proc. Natl. Acad. Sci. USA 92, 6200-6204; Newbert, R.W., Barton, B., Greaves, P., Harper, J., Turner, G., 1997. Analysis of a commercially improved Penicillium chrysogenum strain series: involvement of recombinogenic regions in amplification and deletion of the penicillin biosynthesis gene cluster. J. Ind. Microbiol. Biotechnol. 19, 18-27]. Transcriptional analysis of this amplified region (AR) revealed the presence of at least eight transcripts expressed in penicillin producing conditions. Three of them correspond to the known penicillin biosynthetic genes, pcbAB, pcbC, and penDE. To locate genes related to penicillin precursor formation, or penicillin transport and regulation we have sequenced and analyzed the 56.8 kb amplified region of P. chrysogenum AS-P-78, finding a total of 16 open reading frames. Two of these ORFs have orthologues of known function in the databases. Other ORFs showed similarities to specific domains occurring in different proteins and superfamilies which allowed to infer their probable function. ORF11 encodes a D-amino acid oxidase that might be responsible for the conversion of D-amino acids in the tripeptide L-alpha-aminoadipyl-L-cysteinyl-D valine or other beta-lactam intermediates to deaminated by-products. ORF12 encodes a predicted protein with similarity to saccharopine dehydrogenases that seems to be related to biosynthesis of the penicillin precursor alpha-aminoadipic acid. A deletion mutant, P. chrysogenum npe10 lacking the entire AR including ORF12, shows a partial requirement of L-lysine for growth. ORF13 encodes a putative protein containing a Zn(II)2-Cys6 fungal-type DNA-binding domain, probably a transcriptional regulator. Although some of the ORFs in the AR may play roles in increasing penicillin production, none of the 13 ORFs other than pcbAB, pcbC, and penDE seem to be strictly indispensable for penicillin biosynthesis. The genes located in the P. chrysogenum AR have been compared with those found in the Aspergillus nidulans 50 kb DNA region adjacent to the penicillin gene cluster, showing no conservation between these two fungi. PMID- 16713315 TI - Anoxic-epileptic seizures in Cornelia de Lange syndrome: case report of epileptic seizures induced by obstructive apnea. AB - We describe epileptic seizures including status epilepticus provoked by recurrent obstructive apnea in a child with Cornelia de Lange syndrome. From the age of 10 months, this boy had recurrent respiratory infections with obstructive apnea leading to cyanosis and loss of consciousness. Approximately, 25% of apneas were followed by clonic jerks usually lasting 10 min, but once status epilepticus. He never had unprovoked epileptic seizures. At first he was diagnosed with symptomatic epilepsy and given carbamazepine and phenobarbital, without benefit. Significant improvement occurred after his mother was taught to extract mucus from his upper airways before obstruction occurred. He is no longer on anti epileptic drugs. With this management, he had only one episode of obstructive apnea followed by an epileptic component. The events in this child were anoxic epileptic seizures, that is, epileptic seizures triggered by syncopes. Anoxic epileptic seizures have not previously been described in Cornelia de Lange syndrome. This case illustrates that, even when recurrent epileptic seizures occur in patients with known structural cerebral pathology, the diagnosis of symptomatic epilepsy should not be uncritically accepted. PMID- 16713316 TI - Propofol exerts protective effects on the acute lung injury induced by endotoxin in rats. AB - Acute lung injury (ALI) is a major culprit of mortality in endotoxemia. Propofol has been commonly used in critical ill patients for sedation. This experiment attempted to elucidate the effects and possible mechanisms of propofol on the ALI induced by endotoxin. Experimentations were carried out using anesthetized, ventilated rats and isolated perfused rat lungs. Endotoxemia was induced by intravenous lipopolysaccharide (LPS, 10 mg kg(-1)). Various groups of rats received infusion of physiological saline solution (PSS) and LPS. Five min after LPS, propofol at low dose (5 mg kg(-1)h(-1)) or high dose (10 mg kg(-1)h(-1)) was infused for 6h. In isolated perfused rat lungs, PSS, LPS, and propofol (30 or 60 mg kg(-1)) were added into the perfusion circuit. During or after 6h observation, we determined the lung weight (LW)/body weight ratio, LW gain, exhaled nitric oxide (NO) and protein concentration in broncheoalveolar lavage. Lung pathology was evaluated to quantify the lung injury score. Plasma nitrate/nitrite, methyl guanidine (MG), tumor necrosis factor(alpha), and interleukin-1(beta) were examined. Blood leukocytes were counted. Capillary filtration coefficient (K(fc)) was obtained in isolated perfused lungs. Posttreatment of propofol at low or high dose attenuated or prevented the extent of ALI. It also reduced the plasma nitrate/nitrite, MG, and pro-inflammatory cytokines including tumor necrosis factor(alpha) (TNF(alpha)) and interleukin-1(beta) (IL-1(beta)). In the isolated perfused rat lungs, propofol significantly reduced the LPS-induced increase in K(fc). This agent did not affect the leukocytopenia caused by LPS. Accordingly, the effects of propofol on the ALI were not related to leukocyte activation or sequestration. Our results suggest that propofol exerts protective effect on the endotoxin-induced ALI. The mechanisms of actions may be mediated through inhibition on the release of pro-inflammatory cytokines, free radicals and NO. In addition, propofol abrogates the microvascular leakage of water and protein in the lungs. The results imply that the use of propofol in critically ill is not only for sedation, but also useful for the prevention of inflammatory progression and lung damage. PMID- 16713317 TI - Blood sampling techniques and storage duration: effects on the presence and magnitude of the red blood cell beta-adrenergic response in rainbow trout (Oncorhynchus mykiss). AB - Many teleostean fish, including rainbow trout, regulate red blood cell (RBC) pH (pH(i)) in the presence of a stress-induced acidosis such as hypoxia, hypercapnia, or exhaustive exercise. This is accomplished through activation of RBC Na+/H+ exchange (beta-NHE), ultimately minimizing impairment to oxygen transport. Presence and characterization of the RBC beta-NHE in fish is best tested in blood from cannulated, resting animals; however, several studies have used blood from stressed animals drawn from the caudal vein and stored prior to use. The effects of sampling procedures and storage on the beta-NHE response is not known and is the focus of this study. Whole blood drawn from cannulated, resting rainbow trout was compared with RBCs obtained from the caudal vein rinsed and stored at 4 degrees C for 0, 6, 24, 48, 96 or 144 h. Isoproterenol (10(-5) M), a beta-adrenergic agonist, was added to hypoxia/hypercapnia incubated RBCs in vitro. In all treatments, isoproterenol induced a large beta-NHE response, and storage duration (< or =96 h) had a minimal affect, indicating that rinsing and storing is an easy and viable means by which to obtain RBCs and investigate function. Storage for 144 h still resulted in a significant RBC beta-NHE response; however, viability of RBCs may be compromised. PMID- 16713318 TI - Idiopathic hypertrophic osteopathy in a cat. AB - This report describes the first case of idiopathic hypertrophic osteopathy (HO) in a cat. No causes for the bone pathology were found following evaluation of the physical and laboratory examinations (complete blood count, albumin, creatinine, urea, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase and gamma-glutamyltransferase and urinalysis), and after histopathological evaluation of organs at necropsy. Based on the radiographic, clinical and anatomopathological findings, idiopathic HO was diagnosed. PMID- 16713319 TI - Interstitial nephritis in cats inoculated with Crandell Rees feline kidney cell lysates. AB - Parenteral administration of Crandell Rees feline kidney (CRFK) cell lysates or feline herpesvirus 1, calicivirus, and panleukopenia virus-containing vaccines (FVRCP) grown on CRFK cells induces antibodies against CRFK cells. These antibodies also react with feline renal cell extracts. The purpose of this study was to determine whether interstitial nephritis would be detected in cats that were immunologically sensitized with CRFK lysates, boosted with CRFK lysates, and then biopsied 2 weeks after the booster. Cats (2 per group) were immunologically sensitized against CRFK lysates by administering 10 microg, 50 microg, or 50 microg plus alum 13 times (12 times in the first 50 weeks) over 2 years. Two cats were inoculated three times, 4 weeks apart with an FVRCP vaccine for intranasal administration as kittens, boosted 50 and 102 weeks later, and then renal biopsies taken 2 weeks after the last booster. Neither of the cats vaccinated with the FVRCP for intranasal administration had detectable renal inflammation. One cat in each of the three CRFK lysate sensitization groups had lymphocytic plasmacytic interstitial nephritis. PMID- 16713320 TI - Spontaneous hormonal variations in male cats following gonadectomy. AB - The increased prevalence of obesity after neutering in cats is problematic in veterinary practice. Although many factors seem to be involved, the role of prolactin (PRL) and insulin-like growth factor-I (IGF-I), both implicated in adipose tissue development and glucose intolerance, should be considered. Seven male cats were castrated when 11 months old. Body weight was then recorded for 56 weeks and PRL, IGF-I and leptin assayed for 44 weeks. Body weight increased steadily but only significantly after 36 weeks. It stabilised after 44 weeks, and the cats then gained about 20% of their initial body weight. IGF-I increased rapidly and was significantly higher by week 3. PRL and leptin increased with initial peaks during the eighth and eleventh weeks, respectively. This study confirms that castration rapidly modifies the hormonal balance, partly explaining the body weight increase, and that hormonal changes precede this body weight increase. Hyperleptinaemia is apparently a consequence of excess weight. PMID- 16713321 TI - [Clinical and sonographic diagnosis of occiput posterior position: a prospective study of 350 deliveries]. AB - OBJECTIVE: To evaluate clinical reliability compared to intrapartum ultrasound as a tool to diagnose occiput posterior position and to investigate the proportion of rotations occurring during labour. PATIENTS AND METHODS: 350 women in labor with a singleton fetus in a vertex position were prospectively studied using ultrasound and obstetrical examination. Outcome of labor was also monitored. RESULTS: Reliability of clinical examination is 85,7%, initial occiput posterior position represented 40,2% and most rotated in an anterior position (84, 8%) while only 0,6% of initial anterior positions delivered in occiput posterior position. Logistic regression did not allow to find significant predictor of occiput posterior position rotation. DISCUSSION AND CONCLUSION: Clinical examination is relatively reliable for posterior position diagnosis and in most cases, initially occipitoposterior positions rotate anteriorly. PMID- 16713323 TI - Signalling networks regulating cyclooxygenase-2. AB - Cyclooxygenease-2 (COX-2) is the key enzyme regulating the production of prostaglandins, central mediators of inflammation. The expression of cyclooxygenease-2 is induced by several extra cellular signals including pro inflammatory and growth-promoting stimuli. All signals converge to the activation of mitogen-activated protein kinases (MAPK) that regulate cyclooxygenease-2 mRNA levels both at the transcriptional and post-transcriptional level. The machinery appears to be highly specialized involving activation of distinct signalling molecules depending on the type of extracellular stimulus. Expression of cyclooxygenease-2 mRNA is regulated by several transcription factors including the cyclic-AMP response element binding protein (CREB), nuclear factor kappa B (NFkB) and the CCAAT-enhancer binding protein (C/EBP). Cyclooxygenease-2 is also affected post-transcriptionaly, at the level of mRNA stability. Finally, cyclooxygenease-2 can be affected directly at its enzymatic activity by nitric oxide and nitric oxide synthase (iNOS). Each step of cyclooxygenease-2 regulation can be used as potential therapeutic target. PMID- 16713322 TI - [Evaluation of controlled-release dinoprostone Propess for labor induction]. AB - OBJECTIVE: To evaluate the efficacy of cervix ripening with vaginal controlled release Propess. PATIENTS AND METHODS: A retrospective study of all women who underwent cervical ripening with Propess during the study period from 1(st) January 2002 to 31(st) December 2004 was carried out. A total of 130 patients who experienced Propess was compared with the next following patient who delivered spontaneously matched on gestational age. Modes of delivery, failure of labor, maternal morbidity were recorded. RESULTS: Indications for induction of labor were: post-term pregnancies in 18.5%, pre-eclampsia in 20.8%, oligohydroamnios in 18.5%, post-term pregnancy and oligohydramnios in 10.8%, intra-uterine fetal growth in 6.9%, premature rupture of membranes in 6.9%, diminution of fetal mobility in 6.1% and miscellaneous in 11.5%. Failure of cervical ripening was 21.2%. Patients in the Propess group had a 3.5 fold higher risk of Cesarean section [95% CI: 1.5-8.3; P < 0.04]. There was no case of maternal or fetal death. There was no difference in incidence of maternal complications, and post partum haemorrhage. DISCUSSION AND CONCLUSION: Use of vaginal pessary Propess does not induce adverse maternal or fetal morbidity. However, it was associated with a higher incidence of Cesarean delivery. PMID- 16713324 TI - AOX--a functional marker for efficient cell reprogramming under stress? AB - Functional markers for stress tolerance can be used in plant breeding to identify genotypes with high yield stabilities under various conditions. Thus, a good marker should show a strong correlation with favourable adaptive plant behaviour. The efficient reprogramming of target cells for yield determination is currently considered to be the most important step towards defining abiotic stress tolerance. In this Opinion article, we propose a role for the alternative oxidase (AOX) gene as a marker for genetic variation in cell reprogramming and yield stability. Evidence to support this idea comes from the metabolic role of alternative respiration under stress, the link between AOX activity and differential growth, and the single nucleotide polymorphism recently observed in AOX genes. We propose an innovative, interdisciplinary and global research strategy for future experimentation on AOX genes that could have an application in plant breeding. PMID- 16713325 TI - Is neocortex essentially multisensory? AB - Although sensory perception and neurobiology are traditionally investigated one modality at a time, real world behaviour and perception are driven by the integration of information from multiple sensory sources. Mounting evidence suggests that the neural underpinnings of multisensory integration extend into early sensory processing. This article examines the notion that neocortical operations are essentially multisensory. We first review what is known about multisensory processing in higher-order association cortices and then discuss recent anatomical and physiological findings in presumptive unimodal sensory areas. The pervasiveness of multisensory influences on all levels of cortical processing compels us to reconsider thinking about neural processing in unisensory terms. Indeed, the multisensory nature of most, possibly all, of the neocortex forces us to abandon the notion that the senses ever operate independently during real-world cognition. PMID- 16713326 TI - Seeing it differently: visual processing in autism. AB - Several recent behavioral and neuroimaging studies have documented an impairment in face processing in individuals with Autism Spectrum Disorder (ASD). It remains unknown, however, what underlying mechanism gives rise to this face processing difficulty. One theory suggests that the difficulty derives from a pervasive problem in social interaction and/or motivation. An alternative view proposes that the face-processing problem is not entirely social in nature and that a visual perceptual impairment might also contribute. The focus of this review is on this latter, perceptual perspective, documenting the psychological and neural alterations that might account for the face processing impairment. The available evidence suggests that perceptual alterations are present in ASD, independent of social function. PMID- 16713327 TI - Orthoptic indications for contact lens wear. AB - Orthoptic anomalies are prevalent: they are encountered in at least 5% of patients seen in a typical primary eyecare practice. Several cases are reviewed that highlight the role of contact lenses in treating orthoptic anomalies. In particular, contact lenses are the preferred optical approach to the correction of anisometropia, and it is often argued that anisometropia should be corrected as young as possible. However, fitting contact lenses to patients, particularly children, with anisometropic amblyopia has been problematic because there is no immediate binocular acuity improvement when the contact lenses are inserted which reduces patient motivation. Continuous wear with silicone hydrogels represents a breakthrough for these cases and some illustrative case studies are given. The visual deficit in amblyopia can be reduced in some cases solely by fitting contact lenses, without the need for occlusion therapy. Other orthoptic uses of contact lenses are reviewed including: correcting motor deviations, occlusion, and infantile onset nystagmus. It is concluded that there are orthoptic anomalies where contact lenses are the preferred mode of correction. It is in patients' best interest for practitioners to discuss contact lenses in these cases. PMID- 16713328 TI - Recognition of flowers by pollinators. AB - The flowers of angiosperm plants present us with a staggering diversity of signal designs, but how did this diversity evolve? Answering this question requires us to understand how pollinators analyze these signals with their visual and olfactory sense organs, and how the sensory systems work together with post receptor neural wiring to produce a coherent percept of the world around them. Recent research on the dynamics with which bees store, manage and retrieve memories all have fundamental implications for how pollinators choose between flowers, and in turn for floral evolution. New findings regarding how attention, peak-shift phenomena, and speed-accuracy tradeoffs affect pollinator choice between flower species show that analyzing the evolutionary ecology of signal receiver relationships can substantially benefit from knowledge about the neural mechanisms of visual and olfactory information processing. PMID- 16713329 TI - Nuclear calcium changes at the core of symbiosis signalling. AB - Many plants acquire a significant proportion of their nutrient requirements through mutualistic symbiotic interactions with micro-organisms. Legumes in particular acquire the macronutrients nitrogen and phosphorus, and most likely an array of micro-nutrients, from interactions with nitrogen-fixing bacteria and with mycorrhizal fungi. Although the structures formed to support these interactions are different (nodules compared with arbuscules), there is conservation in early signalling between these two symbioses. It is likely that different receptors for rhizobial or mycorrhizal signals induce responses that feed into a common signalling pathway. In the nodulation signalling pathway, calcium plays an essential role as a secondary messenger, and the component that probably transduces the calcium signal is a unique calcium-activated kinase that is required for both mycorrhization and nodulation. The nodulation signalling pathway contains transcriptional regulators downstream of the calcium-activated kinase that are not required for the mycorrhizal symbiosis. This suggests that different symbiosis-specific signalling pathways are activated downstream of the calcium-activated kinase, and raises the question of how specificity of gene induction can be achieved in two pathways that are both likely to use calcium and a unique calcium-activated kinase to induce different downstream events. PMID- 16713330 TI - Endophyte or parasite--what decides? AB - Symbiosis between a fungus and a plant is a widespread phenomenon in nature. The outcome of such an interaction can vary in a seamless manner from mutualism to parasitism. In most cases, the host plant does not suffer, in fact it often gains an advantage from colonization by a fungus. This benefit is based on a fine-tuned balance between the demands of the invader and the plant response. If the interaction becomes unbalanced, disease symptoms appear or the fungus is excluded by induced host defence reactions. Symbioses of plants with beneficial or neutral endophytes share many common attributes with plant interactions with pathogens. Recent findings emerging from studies of compatible host-fungus interactions have enhanced our understanding of what determines whether the fungus behaves as an endophyte or a parasite and of how plants avoid exploitation by detrimental parasites but benefit from mutualistic endophytes. PMID- 16713332 TI - The tuberous sclerosis genes and regulation of the cyclin-dependent kinase inhibitor p27. AB - Tuberous sclerosis complex (TSC) is an autosomal dominant tumor syndrome that affects approximately 1 in 6000 individuals. It is characterized by the development of tumors, named hamartomas, in the kidneys, heart, skin and brain. The latter often cause seizures, mental retardation, and a variety of developmental disorders, including autism. This disease is caused by mutations within the tumor suppressor gene TSC1 on chromosome 9q34 encoding hamartin or within TSC2 on chromosome 16p13.3 encoding tuberin. TSC patients carry a mutant TSC1 or TSC2 gene in each of their somatic cells, and loss of heterozygosity has been documented in a wide variety of TSC tumors. Recent data suggest that functional inactivation of TSC proteins might also be involved in the development of other diseases not associated with TSC, such as sporadic bladder cancer, breast cancer, ovarian carcinoma, gall bladder carcinoma, non-small-cell carcinoma of the lung, and Alzheimer's disease. Tuberin and hamartin form a heterodimer, suggesting they might affect the same processes. Tuberin is assumed to be the functional component of the complex and has been implicated in the regulation of different cellular functions. The TSC proteins regulate cell size control due to their involvement in the insulin signalling pathway. Furthermore, they are potent positive regulators of the cyclin-dependent kinase inhibitor p27, a major regulator of the mammalian cell cycle. Here we review the current knowledge on how mutations within the TSC genes could trigger deregulation of stability and localization of the tumor suppressor p27. PMID- 16713331 TI - Analysis of chromosome damage in circulating lymphocytes of radiological workers affected by thyroid nodules. AB - The aim of our study was to assess whether or not thyroid nodularity in combination with occupational exposure to low levels of ionising radiation would be correlated with chromosome damage in peripheral lymphocytes. Conventional chromosome-aberration analysis was performed on a group of 92 hospital workers with or without thyroid nodules. On the basis of measurements of their exposure levels, the workers were classified into a low (mean total level=0.03 mSv), medium (mean total level=1.04 mSv) or high (mean total level=8.60 mSv) exposure category. Our results indicate that among workers with thyroid nodules, the high exposed workers showed significantly higher levels of both total (2.35+/-0.34 per 100 cells) and chromosome-type aberrations (1.46+/-0.20 per 100 cells) than medium-exposed (0.98+/-0.42 and 0.68+/-0.25 per 100 cells, respectively) or low exposed workers (1.11+/-0.29 and 0.58+/-0.17 per 100 cells, respectively). Workers without thyroid nodules had comparable frequencies of chromosome aberrations among the three exposure categories. To our knowledge, this is the first study revealing a slight, but significant increase of chromosome damage in peripheral lymphocytes from hospital workers who developed thyroid nodules under conditions of occupational exposure to radiation well below the threshold limit for the workplace. The existence of a possible association between chromosome aberrations and development of thyroid nodularity will be discussed. PMID- 16713333 TI - Elevation of serum riboflavin carrier protein in hepatocellular carcinoma. AB - Early detection of hepatocellular cancer (HCC), makes it surgically resectable with a potential for cure. The test most commonly used to detect HCC is the measurement of serum alpha-fetoprotein (AFP) levels. However, the AFP test is negative in HCC detection in more than 30% of the cases. Riboflavin carrier protein (RCP) is a growth and developmental protein, synthesized and secreted by the liver and hence was of interest to measure its levels in HCC. A prospective double blind evaluation of RCP levels in serum from 93 subjects was undertaken. These included 22 proven cases of HCC, 25 normal controls, 20 cases of alcoholic hepatitis, 20 cirrhotics, and 6 cases of primary biliary cirrhosis (PBC). RCP was measured by a sensitive and specific radioimmunoassay (RIA). RCP was immunohistochemically localized in paraffin sections of liver specimens using standard methods. Mean serum RCP levels in HCC were 21.75+/-14.66ng/ml and were significantly higher (p<0.0001) than those in normal controls (0.73+/-0.25ng/ml), alcoholic hepatitis (1.92+/-0.82ng/ml), PBC (2.16+/-0.74ng/ml), or cirrhosis (5.02+/-1.52ng/ml). Serum RCP levels were elevated in all 22 HCC cases. In contrast serum AFP levels were elevated in 11 of 22 HCC cases. Immunohistochemical analyses revealed positive staining for RCP in liver tumors. We have previously demonstrated elevation of serum RCP levels in breast adenocarcinoma. Our results suggest that serum RCP levels are significantly elevated in HCC also and could potentially serve as a marker for HCC detection under conditions where breast cancer is ruled out. In combination with AFP, serum RCP levels have the potential of serving as a panel of markers for better detection of HCC. PMID- 16713334 TI - Usefulness of elastometry in evaluating the extents of liver fibrosis in hemophiliacs coinfected with hepatitis C virus and human immunodeficiency virus. AB - The newly developed elastometer, FibroScan((R)), was utilized to evaluate liver fibrosis in hepatitis C virus (HCV)- and human immunodeficiency virus (HIV) coinfected 33 hemophiliacs and HIV-uninfected 24 patients with chronic hepatitis C. Chronicity in the liver was categorized into 4 stages by abdominal ultrasound (AUS): 1 (normal or fatty liver); 2 (chronic liver disease, mild); 3 (moderate); and 4 (severe). Stiffness of the liver was significantly increased as AUS stages advanced: 5.4+/-2.2 (N=3) versus 7.5+/-2.7 (N=9), in stage 1; 4.9+/-1.7 (N=2) versus 9.9+/-6.0 (N=10), in stage 2, 13.5+/-4.7 (N=5) versus 12.9+/-5.9 (N=6), in stage 3, and 22.0+/-9.5 (N=14) versus 28.1+/-21.3 (N=8), in stage 4, in non-HIV group and in HIV group, respectively (P=0.004 and 0.007). Stiffness was correlated with AUS stages (r=0.740, P<0.001), platelet counts (PLT; r=-0.642, P=0.001) and 7S domain of type IV collagen (IV-coll; r=0.480, P=0.024) in non-HIV group, while in HIV group, with IV-coll (r=0.801, P<0.001), AUS stages (r=-0.603, P<0.001), procollagen type III peptides (P-III-P; r=0.621, P=0.001), PLT (r= 0.480, P=0.005), and hyaluronic acid (r=0.433, P=0.027). FibroScan((R)) is absolutely noninvasive and can be the alternative to liver biopsy, especially in patients with bleeding tendency. PMID- 16713335 TI - The reappearance of Victoria lineage influenza B virus in Brazil, antigenic and molecular analysis. AB - BACKGROUND: In contrast to influenza A, minor influenza B viruses can co circulate with the dominant strain during an epidemic allowing the re-emergence of old strains and reassortment between those different strains. The 2001-2002 influenza season in the northern hemisphere was distinguished by the re-emergence of the Victoria-lineage viruses, which replaced the Yamagata-lineage, after being restricted to East Asia throughout the 1990s. OBJECTIVES: To describe the antigenic and genetic characteristics of influenza B viruses detected in South and South East Brazil and determine their lineages. STUDY DESIGN: Influenza samples collected during epidemics between 1999 and 2002 were analyzed by indirect immunofluorescence assay (IFA). Positive results were confirmed through multiplex PCR and isolation in cell culture. Isolated viruses were antigenically characterized by hemagglutination inhibition. Fourteen hemagglutinin (HA) gene sequences obtained in this work were used for phylogenetic analysis. RESULTS: Brazilian isolates from 2002 were associated with the Victoria-lineage, diverging from the vaccine used throughout that influenza season in Brazil. CONCLUSIONS: These results indicate the reappearance of Sichuan/7/97-like samples in South and South East Brazilian Regions simultaneously. They indicate the need for neuraminidase gene evaluation and demonstrate the importance of influenza laboratory surveillance to establish which strains should be included in the influenza vaccine. PMID- 16713336 TI - Effect of socioeconomic deprivation on the population risk of incident heart failure hospitalisation: an analysis of the Renfrew/Paisley Study. AB - BACKGROUND: There are few data describing the effect of socioeconomic deprivation on the risk of developing heart failure (HF). AIMS: To examine the relationship between socioeconomic deprivation and hospitalisation with HF over 20 years. METHODS: Between 1972 and 1976, 15,402 individuals, aged 45-64 years, residing in two towns in Scotland, underwent cardiovascular screening. We report hospitalisations with HF over the subsequent 20 years according to Carstairs deprivation category and Social Class. RESULTS: Following screening, 628 men and women (4.1%) were hospitalised with a primary diagnosis of HF. There was a gradient in the risk of HF hospitalisation with increasing socioeconomic deprivation (P=0.003). Of the most deprived individuals, 6.4% were hospitalised for HF compared to 3.5% of the most affluent group. Cox-proportional Hazard models showed that independent of age, sex and baseline risk factors for cardio respiratory status, greater socioeconomic deprivation increased the risk of HF admission (P<0.001, overall). The adjusted risk of admission for HF was 39% greater in the most versus least deprived subjects (RR 1.39 95% CI 1.04-2.01; P=0.04). CONCLUSION: These data show a link between social deprivation and the risk of developing HF, irrespective of baseline cardio-respiratory status and cardiovascular risk factors. PMID- 16713337 TI - Effects of exercise training on cardiac performance, exercise capacity and quality of life in patients with heart failure: a meta-analysis. AB - BACKGROUND: Despite major advances in pharmacological treatment of chronic heart failure (CHF), a number of patients still suffer from dyspnoea, fatigue, diminished exercise capacity and poor quality of life. It is in this context that exercise training is being intensively evaluated for any additional benefit in the treatment of CHF. AIMS: To determine the effect of exercise training in patients with CHF on cardiac performance, exercise capacity and health-related quality of life. A meta-analysis was performed to obtain this goal. METHODS AND RESULTS: After including 35 randomised controlled trials, the methodological quality of each study was assessed, summary effect sizes (SESs) and the concomitant 95% confidence intervals (95% CI) were calculated for each outcome. Quantitative analysis showed statistically significant SESs, at rest, for diastolic blood pressure and end-diastolic volume. During maximal exercise, significant SESs were found for systolic blood pressure, heart rate, cardiac output, peak oxygen uptake, anaerobic threshold and 6-min walking test. The Minnesota Living with Heart Failure Questionnaire improved by an average of 9.7 points. CONCLUSIONS: Exercise training has clinically important effects on exercise capacity and HRQL, and may have small positive effects on cardiac performance during exercise. PMID- 16713338 TI - Familial inflammatory dilated cardiomyopathy. AB - BACKGROUND: Systematic family screening has recently identified dilated cardiomyopathy as an inherited disorder in up to 30% of cases. Mutations in genes encoding proteins responsible for myocardial architecture have been identified, but additional pathophysiological mechanisms including inflammatory reactions have been proposed. AIMS: Identification and characterization of familial DCM, where at least one affected family member fulfils the criteria for inflammatory DCM may lead to a better understanding of the aetiology and pathogenesis of (inflammatory) DCM. METHODS AND RESULTS: Ten families were examined. In six families, clinical characteristics and mode of inheritance were compatible with pure fDCM, fDCM with conduction defect and autosomal recessive fDCM. In four families, (auto-)immune features were diagnosed in affected and non-affected family members. CONCLUSIONS: Familial DCM with an inflammatory component was identified as a specific subgroup of familial DCM. In most cases, the inflammatory process seems to modify, i.e. aggravate, the "classic, cytoskeletopathic" familial DCM, but in some, especially when taking clinical and genetic aspects into account, inflammatory (auto-)immune features can be addressed as the leading pathogenetic principle. Further elucidation of these families may provide a better insight into pathophysiologic processes and may aid in the development of specific therapeutic strategies. PMID- 16713339 TI - Effects of mandibular advancement on upper airway dynamics in awake normal subjects: a pilot study with phrenic nerve stimulation. AB - BACKGROUND: Mandibular advancement (MA) can be an alternative to continuous positive airway pressure (CPAP) in the treatment of obstructive sleep apnea syndrome. This study was designed to describe its effects on upper airway mechanics. METHODS: Six awake healthy subjects (four men, 31+/-8 years, body mass index (BMI) 25+/-2kg/m2) were studied supine. Phrenic stimulation was used to measure pressure-flow relationships in response to diaphragm contractions without pre-activation of upper airway dilators during progressive MA. RESULTS: Phrenic stimulation consistently induced flow limitation (initial peak V'-t1, corresponding to the limiting pressure; reduction to a nadir V'-t2; reincrease to a second peak V'-t3). The upper airway resistances were negatively correlated with MA. Their values at 4mm were significantly lower than at baseline. Further MA reduced resistances. CONCLUSIONS: As with CPAP, MA in normal subjects decreases the propensity of the upper airway to collapse in response to a negative pressure pulse. PMID- 16713340 TI - The influence of age, gender, ethnicity, and insomnia on Epworth sleepiness scores: a normative US population. AB - BACKGROUND AND PURPOSE: This study explored the distribution of Epworth Sleepiness Scale (ESS) scores in a randomly sampled, community population and provided percentile scores that will assist in decision-making in both research and clinical settings. PATIENTS AND METHODS: Participants included 703 individuals between the ages of 20 and 98, with 116 people with insomnia (PWI) and 587 people not having insomnia (PNI). Analyses produced main effects for sleep status and ethnicity. RESULTS: PWI had higher ESS scores than PNI and African-Americans had higher ESS scores than Caucasians, although effect sizes were small. Gender, age group, and season did not impact ESS scores. Receiver operating characteristic (ROC) curve analysis proved the ESS to discriminate poorly between PWI and PNI. CONCLUSIONS: This study found higher percentages of 'sleepy' individuals than previous studies. PWI did have slightly elevated scores on the ESS, but this elevation was not necessarily predictive of an insomnia diagnosis. Results support a continuum of sleepiness/alertness among PWI. PMID- 16713341 TI - Waist circumference predicts the occurrence of sleep-disordered breathing in obese children and adolescents: a questionnaire-based study. AB - BACKGROUND AND PURPOSE: To assess the presence of sleep-disordered breathing (SDB) in a population of obese children and adolescents and to investigate the role of fat distribution in predicting SDB. PATIENTS AND METHODS: One hundred and thirty-two obese children and adolescents, aged 5.0-14.2 years, were consecutively referred to the Department of Pediatrics of the Second University of Naples for screening of obesity. The control group consisted of 453, sex- and age-matched lean subjects selected from local schools in Campania region. The sleep disturbances scale for children (SDSC) questionnaire was used to evaluate SDB prevalence. In all subjects, waist circumference, triceps and sub-scapular skin folds were measured, and Z-scores were calculated. RESULTS: Obese subjects showed significantly higher SDB and sleep hyperhydrosis (SHY) scores than controls. The Z-score of waist circumference correlated with SDB (r=0.32; P=0.006) and SHY factor scores (r=0.37; P=0.005), while the Z-score of body mass index (BMI), triceps and sub-scapular skin folds were not correlated with any SDSC factor scores. Subjects in the higher tertile for Z-score of waist circumference had a significantly higher risk for developing SDB (OR 1.9; 95% IC 1.8-3.2) and SHY (OR 2.1; 95% IC 2.0-4.5). CONCLUSIONS: Waist circumference is a more reliable index than total adiposity and subcutaneous fat in predicting the risk of obese children to develop SDB. PMID- 16713342 TI - Non-positional severe obstructive sleep apnea on polysomnogram became positional OSA with CPAP therapy. PMID- 16713343 TI - An unusual case of episodic stupor. AB - We present a case of episodic stupor associated with a myriad of neuropyschiatric manifestations that baffled doctors until they were recognized as sleep attacks. Continuous monitoring and recognition of a cyclical pattern of symptoms and signs helped to uncover the underlying cause of a rapidly cycling bipolar disorder. The symptoms abated quickly and persistently when treated by olanzapine and lithium. PMID- 16713344 TI - The reliability, validity and responsiveness of the International Restless Legs Syndrome Study Group rating scale and subscales in a clinical-trial setting. AB - PATIENTS AND METHODS: To assess the reliability, validity, and responsiveness of the International Restless Legs Syndrome Study Group's rating scale (the International Restless Legs Scale (IRLS)) (V2.0), using pooled data from two matching, placebo-controlled studies of ropinirole for treating Restless Legs Syndrome (RLS). RESULTS: Pooled patient samples comprised 550 patients in the baseline (validation) sample and 439 patients in the week 12 longitudinal (responsiveness) sample. Factor analysis revealed acceptability of the IRLS total score (accounting for 40% of the variance) and that nine of the 10 IRLS items could also be assigned to two distinct subscales, the symptoms or symptoms impact subscales. The IRLS total score, symptoms and symptoms impact subscales had acceptable construct validity, internal consistency reliability (alpha=0.81, 0.80, and 0.76, respectively), and concurrent validity (r=-0.68, -0.52, -0.70, respectively, with the Restless Legs Syndrome Quality of Life questionnaire (RLSQoL) overall life impact score). IRLS scores differed significantly between different levels of sleep problems and Clinical Global Impression (CGI) of health status (P<0.0001), indicating known groups and clinical validity, respectively. Changes in scores differed significantly among CGI 'global improvement' levels (P<0.0001), providing evidence of responsiveness. CONCLUSIONS: The IRLS total score, symptoms, and symptoms impact subscales are reliable, valid, and responsive in a clinical trial setting. PMID- 16713345 TI - Aryepiglottic fold cyst causing obstructive sleep apnea syndrome. PMID- 16713346 TI - REM sleep-related complete heart block: is it a specific sleep-related disorder? PMID- 16713347 TI - A sleep study with positional hypoxemia. PMID- 16713348 TI - Enhanced external counter pulsation (EECP) for restless legs syndrome (RLS): preliminary negative results in a parallel double-blind study. PMID- 16713349 TI - Social-cognitive correlates of CPAP adherence in experienced users. AB - BACKGROUND AND PURPOSES: Obstructive sleep apnea (OSA) is a condition with serious medical and psychosocial consequences. However, poor adherence with nasal continuous positive airway pressure (CPAP) treatment limits the effectiveness of treatment. Behavior change factors offer one avenue of research to better understand the correlates of CPAP adherence. PATIENTS AND METHODS: Fifty-eight participants who had been diagnosed with OSA and prescribed CPAP treatment agreed to complete questionnaires that assessed sleep apnea symptoms, behavior change factors, and CPAP side effects, and to have their CPAP data downloaded. Behavior change factors from both social cognitive theory (SCT) and the transtheoretical model (TM) were assessed. The primary aim of the study was to examine the relationship between these social-cognitive factors and objectively measured CPAP adherence. RESULTS: Participants had been using CPAP for a mean of 2.1 years. SCT variables (adjusted R(2)=0.115, P=.008) and TM variables (adjusted R(2)=0.157, P<.0001) each accounted for a statistically significant amount of variance in CPAP adherence. CONCLUSIONS: The results suggest that social-cognitive factors may be associated with CPAP adherence in experienced CPAP users. The principal advantage to these theory-driven and empirically validated factors is that they are modifiable and can provide the basis for effective interventions to increase CPAP adherence. PMID- 16713351 TI - Batting with occluded vision: an in situ examination of the information pick-up and interceptive skills of high- and low-skilled cricket batsmen. AB - The capability of cricket batsmen of different skill levels to pick-up information from the pre-release movement pattern of the bowler, from pre-bounce ball flight, and from post-bounce ball flight was examined experimentally. Six highly skilled and six low-skilled cricket batsmen batted against three different leg-spin bowlers while wearing liquid crystal spectacles. The spectacles permitted the specific information available to the batsmen on each trial to be manipulated such that vision was either: (i) occluded at a point prior to the point of ball release (thereby only allowing vision of advance information from the bowler's delivery action); (ii) occluded at a point prior to the point of ball bounce (thereby permitting the additional vision of pre-bounce ball flight); or (iii) not occluded (thereby permitting the additional vision of post-bounce ball flight information). Measurement was made on each trial of both the accuracy of the definitive (forward-backward) foot movements made by the batsmen and their success (or otherwise) in making bat-ball contact. The analyses revealed a superior capability of the more skilled players to make use of earlier (pre bounce) ball flight information to guide successful bat-ball interception, thus mirroring the greater use of prospective information pick-up by skilled performers observed in other aspects of batting and in other time-constrained performance domains. PMID- 16713350 TI - When is sleepiness a disease? How do we measure it? PMID- 16713352 TI - Time for sport and health to be formally linked in a positive way. AB - Preventing sports injury has rarely been cited as an appropriate action to respond to the obesity epidemic, and in fact a recent letter has suggested that those playing sport are as responsible for their predicament as those who are obese. This opinion piece argues that it is time for better prevention and management of sports injury to be seen as part of the complex solution to preventing obesity, rather than being a self-inflicted problem that governments should continue to ignore. PMID- 16713353 TI - Pre-operative preparation for cardiac surgery utilising a combination of metabolic, physical and mental therapy. AB - Cardiac surgery represents major metabolic, physical and mental stresses associated with an increased production of reactive oxygen species. These stresses may hamper post-operative recovery, increasing hospitalisation times and operative mortality. We conducted a quality assurance and feasibility study to evaluate and monitor the safety and efficacy of a new program of combined pre operative metabolic (enhanced antioxidant), physical and mental therapy to counter these stresses prior to cardiac surgery. METHODS: Sixteen cardiac surgery patients received metabolic therapy consisting of the antioxidants coenzyme Q(10) (CoQ(10)) (300 mg) and alpha-lipoic acid (300 mg), combined with magnesium orotate (1200 mg), and omega-3 fatty acids (3g) given daily for a mean 36+/-7 days up until the day of operation. Patients also received a regimen of physical therapy incorporating non-exhaustive, light exercise and stretching techniques. Mental therapy in the form of stress reduction, relaxation and music was also provided. Blood levels of CoQ(10) and malondialdehyde (MDA) were measured and a quality of life (QoL) questionnaire (SF-36) was administered before, after the program and 1 month after surgery. A patient satisfaction survey was conducted at six weeks post-operatively. RESULTS: During the pre-operative period, treated patients (n=16) showed significant improvements in QoL composite scores, physical (33.5+/-4.1 to 41.0+/-4.5, p=0.005) and mental (44.3+/-4.5 to 54.1+/-5.3, p=0.006). CoQ(10) levels increased from 725.6+/-96.1 nmol/l to 3019.9+/-546.4 nmol/l (p=0.006), MDA levels decreased from 2.2+/-0.9 microM to 1.4+/-0.7 microM (p=0.013) and systolic blood pressure decreased from 140+/-4.0 mmHg to 132+/-3.0 mmHg (p=0.002). One month after surgery the treated group (n=14) demonstrated significant improvements from pre-operative baseline in QoL composite scores, physical (38.9+/-4.0 to 57.9+/-5.4, p=0.01) and mental (50.3+/-5.6 to 69.3+/-4.8, p=0.03) compared to a previously reported similar group of cardiac surgery patients (n=74) whose physical and mental scores decreased from 43.0 to 42.8 (p=0.05) and 53.8 to 49.8, respectively (p=0.05). CONCLUSION: These preliminary results suggest that a program of combined metabolic, physical and mental preparation before cardiac surgery is safe, feasible and may improve quality of life, lower systolic blood pressure, reduce levels of oxidative stress and thus has the potential to enhance post-operative recovery. PMID- 16713354 TI - Caseous calcification of the mitral valve ring. AB - Mitral annular calcification (MAC) is found in 10% of patients older than 50 years of age, and represents deposition of calcium between the basal infero lateral ventricular wall and the posterior leaflet of the mitral valve. Caseous calcification, formed from a mixture of calcium, cholesterol and fatty acids, is an uncommon variant found on echocardiography in 0.6% of patients with MAC. The characteristic echographic appearance is of a large echo-dense structure with echo-lucent center and smooth borders. Caseous MAC carries a benign prognosis despite its impressive appearance and should not be misdiagnosed as a more serious condition such as myocardial abscess or tumor. PMID- 16713356 TI - Understanding and exploiting the endogenous interleukin-10/STAT3-mediated anti inflammatory response. AB - Interleukin (IL)-10 performs an irreplaceable role in negatively regulating inflammation, primarily through a mechanism that selectively blocks the expression of pro-inflammatory genes encoding cytokines, chemokines, cell-surface molecules and other molecules involved in the propagation of inflammation. Not surprisingly, IL-10 has attracted interest as a tool to regulate inflammatory diseases. The clinical use of IL-10 as an anti-inflammatory agent has, however, not met expectations. Nevertheless, the signaling pathway used by the IL-10 receptor to generate the anti-inflammatory response is only beginning to be understood and could be a way to regulate inflammation by pharmacological agents. PMID- 16713357 TI - The control of schistosomiasis and soil-transmitted helminths in East Africa. AB - As a result of support from the Bill and Melinda Gates Foundation, schistosomiasis and intestinal or soil-transmitted helminth infections have been the subject of national control programmes in three Eastern and Southern African countries: Uganda, the United Republic of Tanzania and Zambia. Here, we review the significant progress made in their control efforts and highlight the different approaches being adopted to ensure programme effectiveness and sustainability. Although a positive start has been made to reduce morbidity resulting from schistosomiasis and soil-transmitted helminth infections in these countries, it is imperative that support is identified to sustain the programmes until these infections are no longer a public health problem and children can therefore be given a healthy start to life. PMID- 16713358 TI - The Innovative Vector Control Consortium: improved control of mosquito-borne diseases. AB - Few new insecticides have been produced for control of disease vectors for public health in developing countries over the past three decades, owing to market constraints, and the available insecticides are often poorly deployed. The Innovative Vector Control Consortium will address these market failures by developing a portfolio of chemical and technological tools that will be directly and immediately accessible to populations in the developing world. The Bill and Melinda Gates Foundation has supported this new initiative to enable industry and academia to change the vector control paradigm for malaria and dengue and to ensure that vector control, alongside drugs, case management and vaccines, can be better used to reduce disease. PMID- 16713359 TI - A critical analysis of the potential contribution of Orem's (2001) self-care deficit nursing theory to contemporary coronary care nursing practice. AB - BACKGROUND: Over the last three decades, nurse theorists have strongly advocated the use of conceptual models of nursing to guide nursing practice. Within coronary care units, conceptual model-based care has had inconsistent application and is currently challenged by contemporary approaches such as pathways of care. AIMS: This paper aims to critically analyze the use of Orem's self-care deficit nursing theory as a modus operandi to effectively meet the needs of hospitalized patients in coronary care. RESULTS: Although complex both in the language and construction, the self-care model , provides a comprehensive and holistic approach to the care of people in coronary care. CONCLUSIONS: This paper highlights the potential contribution the application of the self-care deficit nursing theory to the coronary care setting from a philosophical and practical perspective. Orem's conceptual model of nursing and current practice in coronary care units share certain similarities that render a useful model for use in practice. However, while it is recommended for consideration for use in both practice and educational settings, further empirical work is required in the area, together with realistic and practical application of the theory to practice in a way that embraces contemporary notions. PMID- 16713361 TI - Knowledge, attitudes, behaviors, and practices regarding epilepsy among Zambian clerics. AB - BACKGROUND: Epilepsy carries a high burden of social morbidity. An understanding of who propagates stigma and the determinants of stigmatizing attitudes is needed to develop effective interventions. Clerics represent an especially influential social group in Africa. Therefore, we conducted a survey of the knowledge, attitudes, behavior, and practices of Zambian clerics with respect to epilepsy. METHODS: We studied clerics in one large rural region as well as in the capital city. The rural survey was conducted door-to-door. In the urban areas, central administration for multiple denominations assisted in survey delivery. The survey, adapted from previously published instruments, included cleric-specific questions and demographic data. Composite scores for knowledge and tolerance were developed. Determinants of higher knowledge and tolerance were assessed. RESULTS: Almost all Zambian clerics know someone with epilepsy and have witnessed a seizure. More than 40% report having a family member with epilepsy. Unfortunately, this familiarity is not associated with more knowledge or tolerance for the condition. Younger clerics, urban dwellers, those with fewer children, and those with more years of formal education were significantly more tolerant. More knowledgeable clerics are more likely to recommend that a person with epilepsy seek care from a physician rather than a traditional healer. Formal education was the most important factor in determining tolerance toward epilepsy. CONCLUSIONS: Zambian clerics are very familiar with epilepsy, yet have relatively little knowledge of the etiology. Many view traditional healers as the appropriate care provider for epilepsy. To decrease stigma and improve the quality of advice offered by clerics to their congregations, educational programs focusing on the biomedical nature of the disorder are needed, particularly in rural regions. PMID- 16713360 TI - Robust systemic transduction with AAV9 vectors in mice: efficient global cardiac gene transfer superior to that of AAV8. AB - It has been recently shown that recombinant adeno-associated virus serotype 8 (rAAV8) is a robust alternative serotype vector that overcomes many of the limitations of rAAV2 and transduces various tissues efficiently and globally through systemic vector administration. AAV9 is a serotype newly isolated from human tissues, but our knowledge of the biology of rAAV9 in vivo is currently limited. Here, we demonstrate by a series of comprehensive side-by-side experiments with rAAV8 and 9 vectors delivered via different routes or at various doses in mice that rAAV9 vectors share the robustness of rAAV8, i.e., (1) very high liver transduction efficiency irrespective of whether vectors are administered intravascularly or extravascularly and (2) substantial transduction in the heart, skeletal muscle, and pancreas by peripheral vein injection. Importantly, rAAV9 transduced myocardium 5- to 10-fold higher than rAAV8, resulting in over 80% cardiomyocyte transduction following tail vein injection of as low as 1.0 x 10(11) particles per mouse. Thus rAAV9, as well as rAAV8, is a robust vector for gene therapy applications and rAAV9 is superior to rAAV8 specifically for cardiac gene delivery by systemic vector administration. PMID- 16713362 TI - Behavioral and antiepileptic effect of acute administration of the extract of the aquatic plant Echinodorus berteroi (Sprengel) Fassett (upright burhead). AB - Echinodorus berteroi is an aquatic plant found in Central America and the Caribbean to which antiepileptic action has been attributed by folk medicine. The aim of the present study was to investigate the potential behavioral and antiepileptic effect of decoctions (1, 5, and 30%, intraperitoneally) of the dried roots. One and five percent decoctions produced hypoactivity in mice. Hyperactivity induced by amphetamine (3mg/kg, subcutaneously) was significantly reduced by the 30% decoction, in rats. The extracts did not modify the latency to the first clonic convulsion or the survival time of isoniazid (210 mg/kg, ip) treated mice. The 30% decoction significantly increased the latency to the first picrotoxin-induced clonic convulsion (7 mg/kg, intraperitoneally), as well as survival time. Repeated administration of the 5% decoction (30-minute intervals) significantly reduced the amplitude (muV) of the epileptic spikes induced by topical application of penicillin to sensorimotor cortex, in curare-treated rats. In summary, the root decoctions of E. berteroi paradoxically exhibited neuroleptic and antiepileptic actions. Nevertheless, these results partly justify the use of the plant for the treatment of epilepsy by practitioners of folk medicine. PMID- 16713364 TI - Can PPARgamma agonists have a role in the management of obesity-related hypertension? AB - Peroxisome proliferator-activated receptors (PPAR) are ligand-activated transcription factors belonging to the nuclear hormone receptor superfamily. PPARgamma is the most extensively studied amongst the three subtypes (alpha, delta and gamma). This receptor is a key modulator of lipid and glucose homeostasis and is predominantly expressed in adipose tissue. Expression of PPARgamma is also found in non-adipose tissues including heart, kidney, spleen, and interestingly, in all relevant components of the vasculature: endothelial and smooth muscle cells. These receptors may therefore also play a role in the regulation of vascular tone and blood pressure. Genetic variants of PPARgamma have also been associated with features of the metabolic syndrome, including obesity and increased blood pressure. The discovery of synthetic ligands for PPARgamma, the Thiazolidinediones (TZDs) has greatly enhanced our understanding of their ligand dependent activation and more importantly their role in vascular pathobiology. Approximately 10 years ago, serendipitous animal experiments demonstrated that despite causing sodium retention, the TZDs actually lowered blood pressure. This review will highlight the role of TZDs in various models of hypertension and discuss their potential role in the management of obesity related hypertension. PMID- 16713363 TI - The relationship of regional frontal hypometabolism to executive function: a resting fluorodeoxyglucose PET study of patients with epilepsy and healthy controls. AB - Executive dysfunction is common in patients with frontal lobe damage and may depend on the location of pathology within the frontal lobes. However, it is unclear how specific brain regions contribute to different aspects of executive functioning. Eighteen patients with frontal lobe epilepsy, 10 patients with juvenile myoclonic epilepsy, and 14 controls completed a series of tests that measure a broad range of executive functions. Resting fluorodeoxyglucose positron emission tomography scans were collected and regional cerebral rates of glucose uptake values were regressed on test scores. Results revealed that frontal lobe metabolic values were strong predictors of executive functioning in patients with epilepsy, but not in healthy controls. However, nonfrontal regions also contributed unique variance on several measures, suggesting that (1) a network of frontal and nonfrontal regions subserve many executive functions and (2) resting hypometabolism can be a useful predictor of executive dysfunction in patients with epilepsy. PMID- 16713365 TI - Does rapid dose titration affect the hepatic safety profile of Bosentan? AB - RATIONALE: Bosentan, a dual endothelin receptor antagonist, has proven efficacy in pulmonary hypertension. Due to an association with hepatic dysfunction, it is typically initiated at a sub-therapeutic dose for 4 weeks before titration to a therapeutic dose. At our institution some patients have undergone rapid titration, to potentially benefit from therapy earlier. This study assesses the impact of this practice on hepatic safety. METHOD: All patients initiated on bosentan therapy before April 2005 were included. Rapidly titrated patients achieved a therapeutic dose by 3 days, whereas standard titration patients were titrated at 4 weeks. All patients were monitored with monthly liver function tests. RESULTS: 149 patients commenced bosentan, of which 55 were rapidly titrated. At baseline, the two groups were similar in age, BMI, diagnosis, 6-min walking distance, alanine aminotransferase (ALT), cardiac index and pulmonary artery pressures. The rapid group had elevated right atrial pressures (9.7 mm Hg versus 7.4 mm Hg, p = 0.016) and worse WHO functional class (p = 0.008) and included less females (31% versus 69%, p = 0.024). The incidence of hepatic dysfunction in all patients was 12.8% at 12 months. There was no statistical difference in incidence between the rapid and standard groups (4% versus 11% at 3 months, p = 0.211 and 6% versus 15% at 12 months, p = 0.219). Of all patients on bosentan, hepatic dysfunction was most significantly associated with a higher baseline ALT (p = 0.021), female sex (p = 0.003) and underlying connective tissue disease (p = 0.025). Subgroup analysis suggested these factors were not confounders when comparing rapid and standard titration. CONCLUSIONS: Rapid and standard titration of bosentan resulted in similar hepatic safety profiles. Baseline ALT, female sex and the presence of connective tissue disease increased the risk of hepatic dysfunction independent of the titration method used. PMID- 16713366 TI - ACE inhibition restores the vasodilator potency of the endothelium-derived relaxing factor, L-S-nitrosocysteine, in conscious Spontaneously Hypertensive rats. AB - OBJECTIVE: The major aim of this study was to determine whether the angiotensin converting enzyme (ACE) inhibitors, captopril or enalapril, restore the diminished vasodilator potency of the endothelium-dependent agonist, acetylcholine (ACh), and the endothelium-derived relaxing factor (EDRF), L-S nitrosocysteine (L-SNC), in conscious Spontaneously Hypertensive (SH) rats. METHODS: The hemodynamic responses elicited by i.v. injections of ACh, L-SNC, and nitric oxide donors such as MAHMA NONOate, were determined in SH rats treated for 7 days with captopril, enalapril, or the direct vasodilator hydralazine. The effects of captopril, enalapril or hydralazine on oxidant stress levels in blood serum and aorta of WKY and SH rats were also determined. RESULTS: Captopril, enalapril and hydralazine elicited equivalent falls in mean arterial pressure and systemic vascular resistances in SH rats. ACh- and L-SNC-induced vasodilation were increased in captopril- or enalapril-treated SH rats such that the responses were equal to those in normotensive Wistar Kyoto rats. The attenuated responses of ACh and L-SNC in SH rats were not improved by hydralazine. The vasodilator effects of MAHMA NONOate, which were substantially augmented in SH rats, were not affected by captopril, enalapril or hydralazine. The levels of oxidant stress were markedly reduced in captopril- or enalapril-treated but not hydralazine treated SH rats. CONCLUSIONS: The finding that the ACE inhibitors improved the vasodilator potencies of L-SNC and the EDRF released by ACh in SH rats, suggests that the diminished vasodilator potency of these compounds was due to augmented ACE activity, which increased oxidant stress levels. This study provides the first evidence to support the concept that ACE inhibition lowers arterial pressure in SH rats, at least in part, by restoring the vasodilator potency of endothelium-derived L-SNC. PMID- 16713367 TI - Immunomodulatory role of Withania somnifera root powder on experimental induced inflammation: An in vivo and in vitro study. AB - The aqueous suspension of Withania somnifera root powder was investigated for their in vivo and in vitro immunomodulatory properties. W. somnifera showed potent inhibitory activity towards the complement system, mitogen induced lymphocyte proliferation and delayed-type hypersensitivity reaction. Administration of W. somnifera root powder did not have a significant effect on humoral immune response in rats. Our results report immunosuppressive effect of W. somnifera root powder, thus it could be a candidate for developing as an immunosuppressive drug for the inflammatory diseases. PMID- 16713368 TI - Effects of pre- and postnatal protein deprivation and postnatal refeeding on myenteric neurons of the rat small intestine: a quantitative morphological study. AB - We investigated weight gain, the size of the small intestine and numbers and sizes of enteric neurons in rats whose mothers had been deprived of protein during pregnancy and who themselves were deprived postpartum. Postnatally, protein deprivation was for 42 days, or for 21 days with refeeding for a further 21 days. Control animals received normal nourishment. Neurons were located by nicotinamide adenine dinucleotide (NADH) diaphorase staining, by acetylcholinesterase (AChE) activity and immunoreactivity for choline acetyltransferase (ChAT). The collagen and elastic fibers in the myenteric ganglia were evaluated histologically. The myenteric ganglia were regular and uniform in the nourished and refed groups. In the undernourished group, the myenteric ganglia were irregularly arranged and the cytoplasm of most of the neurons showed less intense staining for NADH diaphorase, AChE and ChAT. AChE activity and ChAT immunoreactivity showed that most ganglionic neurons were stained in nourished and refed groups, but the neurons of undernourished rats were unstained or moderately stained. The distribution of the connective tissue of the ganglionic capsule was similar in the three groups. There was a decrease in weight of undernourished rats, which was restored in refed rats. The size of the small intestine of the undernourished group was smaller than in the normally fed group, by about 45%, but it was similar in nourished and refed rats. After 42 days of protein deprivation the numbers of neurons that were revealed by NADH diaphorase were fewer than in well nourished rats, but numbers were not different between nourished and refed rats. These observations indicate that protein deprivation alters histological features and acetylcholinesterase activity of neurons and also reduces body weight but these were restored by refeeding. PMID- 16713369 TI - Baroreflex responses in neuronal nitric oxide synthase knoukout mice (nNOS). AB - The present study sought to determine the influence of the neuronal isoform of nitric oxide synthase (nNOS) on the baroreflex gain (mean index: bpm/mm Hg) and on the respiratory response to baroreflex activation by using nNOS knockout (nNOS /-) mice. Experiments were performed with nNOS+/+ wild type (WT; n=12) and nNOS-/ mice (n=11), both of the C57Bl6 strain, randomly assigned to the two groups. Carotid artery and external jugular vein were cannulated under halothane, and after recuperation, urethane was intravenously injected. Baroreflex gain was evaluated by intravenous injection of phenylephrine (PE: 15-18 microg/kg) and sodium nitroprusside (SNP: 46-50 microg/kg) in WT and nNOS-/- anesthetized mice. At baseline, nNOS-/- mice had a comparable heart rate (604.5+/-23.6 vs. 618.7+/ 11.2 bpm) but higher mean arterial pressure (112.4+/-6.3 vs. 94.8+/-3.9 mm Hg, P<0.05) than WT mice. Heart rate reflex was significantly reduced (P<0.05) in nNOS-/- mice, tachycardic and bradycardic responses were -1.04+/-0.3 and -2.03+/ 0.5 in nNOS-/- mice vs. -4+/-0.7 and -4.52+/-0.2 in WT mice, respectively. To characterize the effect of arterial pressure changes on respiratory output, PE was injected in a separate group of WT (n=7) and nNOS-/- (n=6) mice. Tidal diaphragm activity decreased in WT mice (P<0.05); however, diaphragm minute respiration and respiratory frequency were not different between the strains of mice. In nNOS-/- group, heart rate reflex by PE injection was significantly reduced (P<0.05). These findings suggest that the absence of nNOS activity leads to an elevation of the baseline blood pressure and also shows the importance of this enzyme on the transmission of baroreflex signals. PMID- 16713370 TI - Autonomic response and Fos expression in the NTS following intermittent vagal stimulation: importance of pulse frequency. AB - Chronic intermittent stimulation of the vagus nerve (VNS) is an approved adjunctive therapy of refractory epilepsy. Nevertheless, the circuits triggered by VNS under the variable conditions used in patients are not well understood. We analyzed the effect of increasing pulse frequency on physiological variables (intragastric pressure, cardiac and respiratory frequencies) and neuronal activation in the solitary tract nucleus (NTS), the entry level of peripheral vagal afferents, in the rat. For this purpose, we compared the subnuclear distribution of Fos-like immunoreactivity within the NTS following VNS at frequencies selected for their low (1 Hz) or high (10 Hz) therapeutic efficacy. In addition, NADPH diaphorase histochemistry was conducted in double-labeling experiments to check whether activated neurons may express nitric oxide (NO). We demonstrated that increasing pulse frequency had a major influence on the cardiorespiratory response to VNS and on the amount of activated neurons within NTS subdivisions engaged in cardiorespiratory control. These data, in line with clinical observations, suggested that within the range of therapeutic frequency, VNS may favor the regulation by vagal inputs of cortical activities within limbic areas involved in both epileptogenesis and cardiorespiratory afferent control. Furthermore, we did not find any evidence that anticonvulsant VNS might trigger NOergic neurons in the NTS. PMID- 16713371 TI - BDNF and trkB mRNA expression in the hippocampus and temporal cortex during the human lifespan. AB - Brain-derived neurotrophic factor (BDNF) and its receptor tyrosine kinase B (trkB) influence neuronal survival, differentiation, synaptogenesis, and maintenance. Using in situ hybridization we examined the spatial and temporal expression of mRNAs encoding these proteins during diverse stages of life in the human hippocampus and inferior temporal cortex. We examined six postnatal time points: neonatal (1-3 months), infant (4-12 months), adolescent (14-18 years), young adult (20-24 years), adult (34-43 years), and aged (68-86 years). Within the hippocampus, levels of BDNF mRNA did not change significantly with age. However, levels of both the full-length form of trkB (trkB TK+) mRNA and the truncated form of trkB (trkB TK-) decreased over the life span (p < 0.05). In the temporal cortex, BDNF and trkB TK+ mRNA levels were highest in neonates and decreased with age (r = -0.4 and r = -0.7, respectively, both p < 0.05). In contrast, TrkB TK- mRNA levels remained constant across the life span in the temporal cortex. The peak in both BDNF and trkB TK+ mRNA expression in the neonate temporal cortex differs from that previously described for the frontal cortex where both mRNAs peak in expression during young adulthood. The increase in BDNF and trkB TK+ mRNA in the temporal cortex of the neonate suggests that neurotrophin signaling is important in the early development of the temporal cortex. In addition, since BDNF and both forms of its high affinity receptor are expressed throughout the development, maturation, and aging of the human hippocampus and surrounding neocortex they are likely to play roles not only in early growth but also in maintenance of neurons throughout life. PMID- 16713372 TI - The temporal and spatial expression pattern of myosin Va, Vb and VI in the mouse ovary. AB - There are 16 classes of unconventional myosins. Class V myosins have been shown to be involved in transporting cargo to and from the cell periphery. Class VI myosins have also been shown to transport cargo from the cell periphery, although it seems that these proteins have many roles which include the mediation of cell migration and stereocillia stabilisation. With the requirement of myosin VI for Drosophila oogenesis, the localised expression of Myosin V in the developing egg chamber and recent mounting evidence which links myosin VI to the migration of human ovarian cancer cell lines, we wanted to investigate the expression pattern of these two myosin classes in the normal mouse ovary. Here we show that these myosins are expressed, localised and regulated within the oocyte and granulosa cells of the developing mouse follicle. PMID- 16713373 TI - Virus species and virus identification: past and current controversies. AB - The basic concepts used in virus classification are analyzed. A clear distinction is drawn between viruses that are real, concrete objects studied by virologists and virus species that are man-made taxonomic constructions that exist only in the mind. Classical views regarding the nature of biological species are reviewed and the concept of species used in virology is explained. The use of pair-wise sequence comparisons between the members of a virus family for delineating species and genera is reviewed. The difference between the process of virus identification using one or a few diagnostic properties and the process of creating virus taxa using a combination of many properties is emphasized. The names of virus species in current use are discussed as well as a binomial system that may be introduced in the future. PMID- 16713374 TI - Enthalpy/entropy driven activation of the first interquinone electron transfer in bacterial photosynthetic reaction centers embedded in vesicles of physiologically important phospholipids. AB - The thermodynamics and kinetics of light-induced electron transfer in bacterial photosynthetic RCs are sensitive to physiologically important lipids (phosphatidylcholine, cardiolipin and phosphatidylglycerol) in the environment. The analysis of the temperature-dependence of the rate of the P(+)Q(A)(-)Q(B)- >P(+)Q(A)Q(B)(-) interquinone electron transfer revealed high enthalpy change of activation in zwitterionic or neutral micelles and vesicles and low enthalpy change of activation in vesicles constituted of negatively charged phospholipids. The entropy change of activation was compensated by the changes of enthalpy, thus the free energy change of activation ( approximately 500 meV) did not show large variation in vesicles of different lipids. PMID- 16713376 TI - Interfacial tension of the lipid membrane formed from lipid-fatty acid and lipid amine systems. AB - Interfacial tension has been determined for phosphatidylcholine-stearic acid and phosphatidylcholine-stearylamine membranes. Phosphatidylcholine, stearic acid and stearylamine were used in the experimental. The interfacial tension values of the pure components are 1.62x10(-3) N/m, - 1.54x10(-2) N/m and 4.40x10(-3) N/m (hypothetical values), respectively. The 1:1 complexes were formed during formation of phosphatidylcholine-stearic acid and phosphatidylcholine stearylamine membranes. The following parameters describing the complexes were determined: the surface concentrations of the lipid membranes formed from these complexes, A(3)(-1), the interfacial tensions of such membranes, gamma(3) and the stability constants of these complexes, K. PMID- 16713375 TI - Low-frequency magnetic field effect on cytoskeleton and chromatin. AB - The effect of magnetic fields on the living systems is studied in vivo or in vitro in very broad spectrum of organisms, cells and tissues. The mechanism of their acting is not known until now. We studied low-frequency magnetic field effect on cytoskeleton and on the structure of chromatin in human cells. We used cell line of small lung carcinoma (A549) and the effects of magnetic field on cytoskeleton and higher-order chromatin structure were analyzed 96 h of magnetic field exposure. Magnetic field generated by the cylindrical soil was homogenous and the cells were cultivated at 37 degrees C in humidified atmosphere containing 5% CO(2). Magnetic field induction was B(m)=2 mT and the net frequency f=50 Hz. In such affected and control cells the F-actin was estimated using FITC conjugated Phalloidin and mitochondria were studied using MitoTracker (Molecular Probes). Images of cytoskeleton and genetic loci were acquired using confocal microscopy and analysis was performed by FISH 2.0 software. Slight morphological changes of F-actin filaments and mitochondria were observed in affected cells and nuclear condensation was found. These effects could be related to the process of cell death apoptosis probably induced by magnetic field. The studies aimed at centromeric heterochromatin (9cen) did not show statistically significant changes. Therefore, we suggest that magnetic field has no influence on higher order chromatin structure but certain changes could be observed on the level of cytoskeleton. However, these statements need a thorough verification. Our preliminary experiments will be extended and the effect of magnetic field on another structures of cytoskeleton and cell nuclei will be further studied. PMID- 16713377 TI - Ion-exchange voltammetry of dopamine at Nafion-coated glassy carbon electrodes: quantitative features of ion-exchange partition and reassessment on the oxidation mechanism of dopamine in the presence of excess ascorbic acid. AB - The incorporation/exclusion features of dopamine (DA), ascorbic acid (AA) and uric acid (UA) are evaluated for Nafion (NA)-coated glassy carbon electrodes (GCE) of different thicknesses. The ion-exchange partition of DA(+) between the NA film and the sodium phosphate electrolyte is evaluated by determining the partition coefficient (k(D)) and the apparent diffusion coefficient (D(app)) in thick NA films which were 401 and 1.5 x 10(-9) cm(2) s(-1), respectively. The solution diffusion coefficient was found to be 6.0 x 10(-6) cm(2) s(-1). Also, the effect of NA loading and of the voltammetric timescale on DA voltammetry in the presence of excess AA is assessed, at physiologic like conditions. It is demonstrated that, although AA is excluded at the NA coating, a catalytic regeneration of DA, induced by AA, occurs at the interface NA film/electrolyte resulting from the diffusion of the o-quinone product of DA oxidation from the electrode surface to that interface. The interference of AA in the voltammetric signal of DA is eliminated using 18 microg mm(-2) NA films and v> or =0.5 V s( 1). Therefore, fast, selective and sensitive voltammetric analysis of DA at concentrations<100 microM in the presence of excess AA, e.g., 1 mM is achieved. PMID- 16713378 TI - Conformational change of membrane proteins leads to shape changes of red blood cells. AB - High-resolution atomic force microscopy (AFM) allows a new insight into the surface of mammalian cells. Using the human red blood cell (RBC) as a model, we have demonstrated an important correlation between the conformation of membrane proteins measured from the external face of the cell and the cell shape. PMID- 16713380 TI - On the temperature dependence of the dielectric membrane properties of human red blood cells. AB - Electrorotation (ER) spectra of human red blood cells (HRBCs) have been recorded in the frequency range from 10 kHz to 250 MHz in a 4-electrode microchip chamber. The cells were suspended at conductivities in the range from 0.02 to 3.00 S/m (corresponding to an ionic strength range from 1.6 to 343 mM) at temperatures between 10 degrees C and 35 degrees C. Generally, the characteristic frequencies as well as the rotation speeds of the first (membrane-dispersion) and second ER peaks increased with temperature. The rotation speed increase was largely correlated to the temperature dependence of the medium viscosity. Standard temperature dependencies were assumed for the conductivities and permittivities of cytoplasm, membrane, and external solution to explain the frequency shifts, starting from the cell parameters of Gimsa et al. [Gimsa et al., 1996, Biophys. J. 71: 495-506.]. The membrane capacitance was assumed to be temperature independent, based on the permittivity of alkyl-chains. Under these assumptions, the spectra could be well fitted only in a narrow temperature range around 20 degrees C. The temperature dependence of the first characteristic frequency was much stronger than predicted. In addition, around 15 degrees C, an anomalously high rotation speed was observed for the first peak at low external conductivities. Interestingly, this finding corresponds to the change in the chloride transport rate described by Brahm [Brahm, 1977, J. Gen. Physiol. 70: 283 306.]. PMID- 16713381 TI - Effect of electromagnetic fields on the denitrification activity of Paracoccus denitrificans. AB - Enzymatic activity (denitrification) of Paracoccus denitrificans was estimated electrochemically by reduction of duroquinone (DQ). Graphite electrodes covered with whole bacterial cells behind a dialysis membrane were used for measurement. P. denitrificans reduce nitrate and/or nitrite under anaerobic conditions to nitrogen gas. DQ acts as an electron mediator. After donation of the electrons to the respiratory system of the bacteria, produced DQ is reduced to durohydroquinone on the electrode surface electrocatalytically. P. denitrificans were exposed to low-frequency magnetic field (10 mT, 50 Hz) for 24 min. In comparison with the control samples, the reduction peak of I-E curves that represent denitrification activity of the cells decreased significantly after magnetic field exposure. The decrease of the peak current was about 20%. The CFU colony forming units-method was used to estimate the number of surviving bacteria. After 24 min exposure of 10 mT magnetic field P. denitrificans culture on electrode indicates 21% bacterial death. PMID- 16713382 TI - Electrochemical behaviour of 2,8-dihydroxyadenine at a glassy carbon electrode. AB - The electrochemical behaviour of 2,8-dihydroxyadenine (2,8-DHA)- the main adenine oxidation product- has been investigated over a wide pH range at a glassy carbon electrode (GCE) using cyclic, differential pulse and square wave voltammetry. The oxidation of 2,8-DHA is a quasi-reversible process, pH dependent and occurs with the formation of a main oxidation product, P(2,8-DHA), that strongly adsorbs on the electrode surface. The reduction of 2,8-DHA also occurs and is a reversible process in the absence of molecular oxygen. In electrolytes with pH between 4 and 9 two consecutive reversible charge transfer reactions were identified. However, it was observed that O(2) interfered with the reductive electron transfer process of 2,8-DHA and that, in the presence of oxygen, the reduction of 2,8-DHA occurs at less negative potentials than in the absence of oxygen. PMID- 16713383 TI - Effects of low-frequency magnetic fields on the viability of yeast Saccharomyces cerevisiae. AB - A 50 Hz magnetic field effect on the growth of yeasts Saccharomyces cerevisae was studied. The cylindrical coil induced magnetic fields with inductions up to 10 mT. Duration of exposure varied up to 24 min. Exposure took place at laboratory temperature (24-26 degrees C) and the air ventilator maintained the temperature at the place of the sample. We measured the growth curves of yeasts in broth and we calculated the number of CFU (colony forming units) on solid soil. We found that magnetic field decreases the number of yeasts, and slowed down their growth. The result is similar to the experiments with bacteria E. coli, S. aureus and L. adecarboxylata. It seems that the magnetic fields kill a part of yeasts and the bigger part of them survives and continues in their growth. PMID- 16713384 TI - Trans fatty intakes during pregnancy, infancy and early childhood. AB - All of the essential n-6 and n-3 fatty acids accumulated by the fetus must be derived by transfer from the maternal circulation, and ultimately must originate from the maternal diet. After birth, the breast-fed infant receives essential fatty acids via mother's milk, or human milk substitutes and later complementary foods. Trans fatty acids (TFA) may have adverse effects on growth and development through interfering with essential fatty acid metabolism, direct effects on membrane structures or metabolism, or secondary to reducing the intakes of the cis essential fatty acids in either mother or child. TFA are transported across the placenta and secreted in human milk in amounts that depend on the maternal dietary intake. Inverse associations have been shown between TFA and the essential n-6 and n-3 fatty acids in newborn infants, human milk and preschool children. This support the need to reduce industrially produced trans fatty acids (IP-TFA) and improve dietary fat quality, particularly by increasing intake of n 3 fatty acids. PMID- 16713385 TI - A trans world journey. AB - A high intake of industrially produced trans fatty acids (IP-TFA) is associated with increased risk of coronary heart disease (CHD), and a daily intake as low as possible is required to minimize health risks. To achieve this at the individual level in Denmark, legislation limited IP-TFA in foods to a maximum of 2% of fat content from 2004. We assessed the potential exposure of consumers to IP-TFA by analysing popular foods in Denmark, and in 25 other countries. Fifty-five servings of French fries and chicken nuggets, 87 packages of microwave popcorn, and 393 samples of biscuits/cakes/wafers with "partially hydrogenated vegetable fat" listed high on the food label were bought between November 2004 and February 2006. The content of IP-TFA was analysed by standardized methodology. We defined a "high trans menu" as a large size serving of French fries and nuggets, 100 g of microwave popcorn, and 100 g of biscuits/wafers/cakes. The amounts of IP-TFA in a "high trans menu" was 30 g in 2001 in Denmark, but was reduced to less than 1g in 2005. By contrast, a "high trans menu" provided more than 20 g in 17 out of 18 countries, with Hungary, Czech Republic, Poland, Bulgaria, and USA, ranking highest with 42, 40, 38, 37, and 36 g, respectively. The legislation in Denmark has reduced the exposure of IP-TFA at the individual level without noticeable effect on availability, price, and quality of foods previously containing high amounts of IP-TFA. The findings of high concentrations of IP-TFA in popular foods outside Denmark suggest that millions of people inside and outside EU have intakes of IP-TFA that may increase their risk of CHD. The Danish experience demonstrates that this risk can be eliminated. PMID- 16713386 TI - Trans fatty acids and birth outcome: some first results of the MEFAB and ABCD cohorts. AB - Using data from two longitudinal pregnancy and birth cohorts in the Netherlands, the potential influence of trans fatty acids (TFA) on fetal development was investigated by multiple regression analysis. After adjustment for a large number of potential confounders, birth length (BL) and head circumference (HC) were significantly and negatively related to the C18:1trans concentration in phospholipids isolated from umbilical plasma (HC), umbilical arterial walls (HC and BL), or umbilical venous walls (BL). A significant and negative relationship was also observed between birth weight and the C18:1trans concentration in maternal plasma phospholipids at early pregnancy, but this attenuated upon correction for potential confounders. These results indicate that TFA may compromise fetal development. PMID- 16713387 TI - Labeling of trans fatty acid content in food, regulations and limits-the FDA view. AB - With the scientific evidence associating trans fatty acid (TFA) intake with an increased risk of coronary heart disease (CHD), the U.S. Food and Drug Administration (FDA) issued a final rule that requires the declaration of the amount of TFA present in foods, including dietary supplements, on the nutrition label by January 1, 2006. The addition of TFA to the nutrition label will lead to the prevention of 600 to 1200 cases of CHD and 240-480 deaths each year saving Dollars 900 million to Dollars 1.8 billion per year in medical costs, lost productivity, and pain and suffering. For the purpose of nutrition labeling, TFA are defined as the sum of all unsaturated fatty acids that contain one or more isolated (i.e. non-conjugated) double bonds in a trans configuration. There are many issues that FDA has yet to resolve: (1) defining nutrient content claims for "free" and "reduced" levels of trans fat, (2) placing limits on the amount of TFA in conjunction with saturated fat limits for nutrient content claims, health claims, and disclosure and disqualifying levels, (3) a daily value, and (4) a possible footnote or disclosure statement to enhance consumer understanding of cholesterol raising lipids. FDA issued an Advanced Notice of Proposed Rulemaking (ANPR) requesting comments on the unresolved issues. FDA will also be conducting consumer research to determine consumer understanding of various TFA labeling possibilities. Comments to the ANPR, results of consumer research and current science will be used by FDA to resolve these issues and to determine future rulemaking for TFA labeling. PMID- 16713388 TI - Trans fatty acids and insulin resistance. AB - Since trans fatty acids (TFA) might interfere with cell membrane functions, there are reasons to believe that high TFA intakes could affect insulin sensitivity and consequently diabetes risk. It is possible that low amounts of TFA consumed during long time-periods might be clinically relevant. Data from controlled intervention studies investigating the effects of TFA on insulin sensitivity are reviewed. The results show no significant effect of TFA on insulin sensitivity in lean healthy subjects. However, there is some evidence that TFA could impair insulin sensitivity compared to unsaturated fat in insulin resistant or diabetic individuals. This is especially true for conjugated TFA, i.e. conjugated linoleic acid (CLA), which clearly impairs insulin sensitivity. In fact, the effect of CLA on insulin action is the most dramatic adverse effect described for a dietary fatty acid. The inconsistent effect of TFA as a group might partly be due to methodological limitations (e.g. few studies, short duration or small sample size) and differences between studies in design, type and amount of TFA used. Large controlled trials have been required to demonstrate adverse effects of saturated fat on insulin sensitivity, and similar efforts will probably be needed to clarify the effect of TFA on insulin sensitivity. PMID- 16713390 TI - Regulation of trans fats: the gap, the Polder, and McDonald's French fries. AB - Lowering the intake of trans fatty acids (TFA) probably reduces the incidence of coronary heart disease. Estimates of the reduction vary from 4% based on changes in plasma LDL and HDL concentrations alone, to > 20% based on epidemiological associations when TFA intake is lowered by 2% of energy (5 g/day). Even the lowest estimate represents enough cases to justify measures to reduce TFA intake. In The Netherlands, a major reduction in TFA content of retail foods has been achieved in the 1990s through efforts of industry; government intervention has been minimal. Societal pressure is now helping to reduce the TFA content of fast foods. McDonald's French fries in The Netherlands now have less than 4% trans and 24% saturates, as opposed to 21% trans and 21% saturates in the USA. This illustrates the feasibility of reducing TFA in fast foods without increasing saturates. As a result of these developments, dairy and meat have become the major remaining source of TFA in Europe. The question whether these ruminant TFA have the same effect on coronary heart disease risk as industrial TFA has not been settled. PMID- 16713389 TI - Intake of ruminant trans fatty acids and risk of coronary heart disease-an overview. AB - Epidemiological studies have shown a strong direct (positive) association between the intake of trans fatty acids (TFA) and the risk of coronary heart disease (CHD), primarily accounted for by industrially produced TFA (IP-TFA). However, comparisons between ruminant TFA (R-TFA) and IP-TFA and risk of CHD have been based on quintiles of intake, which implies that the associations between the two sources of TFA and the risk of CHD were described across different ranges of intake. Controlled metabolic studies of the effect of intake of total and specific R-TFA on CHD risk factors are warranted. Moreover, further epidemiological studies of intake of R-TFA and risk of CHD in populations with a high intake of R-TFA are warranted. PMID- 16713391 TI - Trans, and n-3 polyunsaturated fatty acids and vascular function-a yin yang situation? AB - Trans fatty acids (TFA) and n-3 polyunsaturated fatty acids (n-3 PUFA) have opposite effects on several biological functions. We report a study on the effects on risk markers for cardiovascular disease. Eighty-seven healthy males were randomly assigned to 8 weeks of daily intake of either 20 g of industrially produced TFA (IP-TFA), 4 g n-3 PUFA, or control fat, incorporated in bakery products as part of the daily food. High-density lipoprotein cholesterol decreased in the TFA-group, triglycerides and mean arterial blood pressure decreased in the n-3 group. Heart rate variability (HRV), arterial dilatory capacity, flow mediated vasodilation, compliance, and distensibility were unchanged. Post hoc, we did a subgroup analysis of the results from the subjects with normal initial HRV. In these, 24-h heart rate (HR) was significantly increased by approximately three beats/min in the TFA group, with a decrease of the same magnitude in the n-3 group. A high HR is associated to an increased mortality and vice versa. Our results thus support the notion that IP-TFA and n-3 PUFA affect risk for cardiovascular mortality via mechanisms not only related to changes in plasma concentrations of lipids and lipoproteins. PMID- 16713392 TI - Should trans fatty acids be viewed as membrane-active drugs? AB - Trans fatty acids (TFA) can modify cellular function by interacting with hydrophobic regions of membrane proteins. Because these interactions resemble those of pharmacological agents, it might be appropriate to view TFA as drugs. PMID- 16713393 TI - Trans fatty acids - effects on systemic inflammation and endothelial function. AB - Consumption of trans fatty acids (TFA) predicts higher risk of coronary heart disease, sudden death, and possibly diabetes mellitus. These associations are greater than would be predicted by effects of TFA on serum lipoproteins alone. Systemic inflammation and endothelial dysfunction may be involved in the pathogenesis of atherosclerosis, acute coronary syndromes, sudden death, insulin resistance, dyslipidemia, and heart failure. Evidence from both observational and experimental studies indicates that TFA are pro-inflammatory. Limited evidence suggests that pro-inflammatory effects may be stronger for trans isomers of linoleic acid (trans-C18:2) and oleic acid (trans-C18:1), rather than of palmitoleic acid (trans-C16:1), but further study of potential isomer-specific effects is needed. TFA also appear to induce endothelial dysfunction. The mechanisms underlying these effects are not well-established, but may involve TFA incorporation into endothelial cell, monocyte/macrophage, or adipocyte cell membranes (affecting membrane signaling pathway relating to inflammation) or ligand-dependent effects on peroxisome proliferator-activated receptor (PPAR) or retinoid X receptor (RXR) pathways. Activation of inflammatory responses and endothelial dysfunction may represent important mediating pathways between TFA consumption and risk of coronary heart disease, sudden death, and diabetes. Further study is indicated to define these effects of TFA and the implications of such effects for cardiovascular health. PMID- 16713394 TI - Trans fatty acids and blood lipids. AB - Intake of trans-unsaturated fatty acids (TFA) has been consistently shown in multiple and rigorous randomized trials to have adverse effects on blood lipids, most notably on the LDL:HDL cholesterol ratio, which is a strong marker of cardiovascular risk. When a mixture of TFA isomers obtained by partial hydrogenation of vegetable oils is used to replace oleic acid, there is a dose dependent increase in the LDL:HDL ratio. The relationship between amount of TFA as % of energy and the increase in the LDL:HDL ratio appears to be approximately linear, with no evidence of a threshold at low levels of intake, and with slope twice as steep as that observed by replacing oleic with saturated fats. The average impact of TFA induced changes in the LDL:HDL ratio correspond to tens of thousands premature deaths in the US alone. Although dramatic, this effect is substantially smaller than the increase in cardiovascular mortality associated with TFA intake in epidemiological studies, suggesting that other mechanisms are likely to contribute to the toxicity of TFA. PMID- 16713395 TI - The scientific basis for trans fatty acid regulations-is it sufficient? A European perspective. AB - The C18:1 trans fatty acids (TFA) comprise a variety of positional isomers, but no definite differences have been documented so far between the metabolic and health effects of industrial and ruminant TFA. In Europe the intake of industrially produced TFA (IP-TFA) has declined, and the majority of TFA are of ruminant origin. TFA have been replaced with cis-unsaturates in soft margarines and they have been reduced also in industrial fats, but often by using palm kernel oils. When modifying the dietary fat composition the proportion of saturated plus TFA should be kept to one-third of total dietary fatty acid intake. PMID- 16713396 TI - World-wide consumption of trans fatty acids. AB - Dietary trans fatty acids (TFA) originate from commercially hydrogenated oils and from dairy and meat fats. Estimates of dietary TFA consumption vary with dietary habits and food supply as well as methods used to estimate consumption. Methods include: (1) market share data, (2) laboratory analysis of duplicate portions or composite diets, (3) analysis of consumption data of a representative population, and (4) biomarkers, such as human milk. In North America, daily intakes have been estimated by food frequency questionnaire to be 3-4 g/person and by extrapolation of human milk data to be greater than 10 g/person. Diets in northern Europe traditionally have contained more TFA than in Mediterranean countries where olive oil is used. Intakes in Europe range from minimal values in Italy, Portugal, Greece and Spain (1.4-2.1 g/day) to greater values for Germany, Finland, Denmark, Sweden, France, United Kingdom, Belgium, Norway, The Netherlands, and Iceland (2.1-5.4 g/day) Recent decreases in dietary TFA have been observed due to modifications of commercial fats and changes in consumer choices. The impact of legislation restricting use of hydrogenated fats and requiring TFA content on food labels awaits future studies. PMID- 16713398 TI - Trans-fatty acids and sudden cardiac death. AB - Sudden cardiac death (SCD) is usually due to ventricular fibrillation and can occur as a first manifestation of heart disease. Prevention of ventricular fibrillation and SCD with n-3 polyunsaturated fatty acids is well documented. Trans-fatty acids (TFA) in the diet and cell membranes might affect the risk of SCD as well. We review evidence from an observational study that high levels of trans-18:2 (9 cis-, 12 trans- and 9 trans-, 12 cis-isomers of linoleic acid) in red blood cell membranes are associated with markedly higher risk of SCD. In contrast, cell membrane levels of trans-18:1 (trans-isomers of oleic acid), the major TFA in foods, do not appear associated with higher risk of SCD. While further studies are needed to investigate possible effects of trans-18:2 on arrhythmia, it would be prudent to limit dietary intake of trans-18:2. PMID- 16713397 TI - The effect of the regulation on trans fatty acid content in Danish food. AB - The content of trans fatty acids (TFA) in Danish food has been monitored for the last 30 years. In margarines and shortenings the content of TFA has steadily declined from about 10 g/100 g margarine in the seventies to practically no TFA in margarines in 1999. In order to efficiently reduce the health risk related to TFA, Denmark decided to impose a maximum level of 2 g/100 g fat on industrially produced TFA (IP-TFA) with the Danish Order no. 160 of March 2003, as labelling was deemed insufficient to protect the consumers, especially risk groups like children or people with high intake of fast foods. A broader range of food was monitored with 253 samples in 2003 and 148 samples in 2005 after the Danish regulation was in effect. The investigations show that the TFA content has been reduced or removed from the products with high TFA content originally, like French fries, microwave oven popcorn and various bakery products, so IP-TFA are now without any significance for the intake of TFA in Denmark. PMID- 16713399 TI - The genetic background of osteoporosis in cystic fibrosis: association analysis with polymorphic markers in four candidate genes. AB - BACKGROUND: Reduced Bone Mass Density (BMD) is frequent in Cystic Fibrosis (CF). Potentially, other genes than the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene may contribute to the bone phenotype variability in CF patients. METHODS: Four candidate genes likely associated with BMD variability were studied: the vitamin D receptor (VDR) gene, the estrogen receptor alpha (ESR1), the calcitonin receptor (CALCR) and the type I alpha 1 collagen (COL1A1) gene. A complete bone and CF evaluation was obtained for 82 subjects (39 m, 43 f): 15 had normal BMD (group 1), 46 were osteopenic (group 2), and 21 were osteoporotic (group 3). RESULTS: No statistical difference was found among the three groups for age, sex, pancreatic status, and vertebral fractures, nor for any of the biochemical markers. Weight, Body Mass Index (BMI), and FEV1, scored significantly worse in the two groups with the lowest T score. The CFTR mutations R1162X and F508del were more frequent in patients with lower BMD (p=0.044 and p=0.071). There was no significant difference in the distribution of the five marker genotypes among the 3 groups defined according to the unadjusted or adjusted (BMI and FEV1) BMD T score. No significant correlation was found between the VDR, CALCR, or COL1A1 gene polymorphisms and reduced BMD values. The individual ESR1 PvuII-XbaI haplotype C-A is associated to elevated u-calcium levels whereas the haplotype T-A is associated to lower values (p=0.00251). CONCLUSIONS: There was no evidence that the genes under study, with the possible exception of ESR1 gene variants, may modulate bone phenotype in CF. PMID- 16713400 TI - Predicting hypoxia in cystic fibrosis patients during exposure to high altitudes. AB - BACKGROUND: For patients with cystic fibrosis (CF)-related partial respiratory insufficiency and reduced arterial oxygen tension at ground level, the mild hypobaric environment on commercial jet aircraft poses the risk of severe hypoxemia. Thus, physicians should be able to estimate the extent of in-flight hypoxia. OBJECTIVES: To derive tools for estimating the expected drop in arterial oxygen partial pressure (paO(2)) and oxygen saturation (saO(2)) in young adult CF patients with mild to moderate airway obstruction during exposure to the hypobaric conditions aboard commercial aircraft and to test the predictive power of a hypobaric chamber simulation. METHODS: Blood gases of 12 CF patients were measured at ground level, at two altitudes in a hypobaric chamber (2000 and 3000 m) and during two 3.5-h flights at cabin altitudes of 1855 m and 1700 m. The altitude dependence of paO(2) and saO(2) in the chamber and during the flights was calculated and results were used to derive estimation equations for in-flight values. RESULTS: In the chamber, saO(2) decreased by 0.33% per 100 m vertical ascent, and this rate increased significantly at altitudes >2000 m. Predicted saO(2) differed from in-flight value by <5%, and agreement between in-flight saO(2) decrease rate and chamber data was good. paO(2) decreased at a rate of 0.99 mm Hg/100 m in the chamber and by 1.33 mm Hg/100 m during flights. None of the subjects showed any clinical symptoms during the flights and the chamber simulation. CONCLUSION: During our worst-case scenario, i.e. the hypobaric chamber simulation at 3000 m, 90% of patients tolerated paO(2) values below the commonly recommended threshold of 50 mm Hg, probably due to adaptation to chronic hypoxemia and lung function impairment. We propose the following equations for an estimation of the expected extent of in-flight hypoxemia in CF patients with mild to moderate airway obstruction and a flight duration of up to 3.5 h: paO2[Alt]=paO2[ground] -1.33 x Alt[mm Hg], and -saO2[Alt]=saO2[ground] -0.33 x Alt [%], with Alt=altitude in 100 m. In addition to the overall clinical situation of a patient, these equations will serve as a practical supportive tool for the assessment of the fitness to fly in the primary care setting. PMID- 16713402 TI - Components to the acid-base related ventilatory drives in the South American lungfish Lepidosiren paradoxa. AB - Lungfish are closely related to terrestrial vertebrates (tetrapoda). Like tetrapods, the South American lungfish (Lepidosiren paradoxa) has central chemoreceptors involved in regulation of acid-base status. However, no data were available on peripheral CO(2)/[H(+)] receptors. Therefore, we tested the hypothesis that such receptors exist by measuring the ventilatory responses during a 5h exposure to combined aquatic/gas phase hypercarbia 7% (approximately 49 mmHg). Normocarbic control ventilation was 22 ml BTPS kg(-1)h(-1), and hypercarbia increased ventilation to 175 ml BTPS kg(-1)h(-1) at 5h. This procedure was repeated with the modification that normocarbic mock CSF (pH 7.45; P(CO2) = 20.7 mmHg) was applied to superfuse the cerebral ventricular system during the last 2h of the experiment. This served to eliminate the hypercarbic stimulus to the central chemoreceptors, while possible responses from peripheral chemoreceptors would remain intact. Peripheral receptors were detected, since ventilation became reduced to 62 ml BTPS kg(-1)h(-1) (P<0.05), which exceeds the initial normocarbic control ventilation (P<0.05). Based on this, the peripheral contribution accounted for 20% of the total response to hypercarbia, similar to the contribution of these receptors in man. PMID- 16713403 TI - Inspiratory muscle endurance testing: pulmonary ventilation and electromyographic analysis. AB - This study analyzed regional pulmonary ventilation and electromyographic (EMG) activity of the respiratory muscles during an inspiratory muscle endurance (IME) test in 10 young women. Radioaerosol (99mTc-DTPA) was generated using a jet nebulizer connected to a linear inspiratory loading system. The lung scintigraphic analysis showed an increase in the radioaerosol deposition using loads of 20 and 30 cmH(2)O (p<0.01). The vertical gradient showed a larger radioaerosol deposition in the medium third of the lungs during the control period (p<0.001). There were larger amounts of radioaerosol deposition in the medium third when compared with the upper and lower third at 30 cmH(2)O (p<0.001). The horizontal gradient showed a larger deposition in the intermediate and central segments during all phases (p<0.00). Electromyographic activity from the muscles of the lower rib cage increased with loads of 20 and 30 cmH(2)O (p<0.03). There was an increase in deposition of radioaerosol when the load increased (r=0.584, p=0.000 for the left lung and r=0.609, p=0.000 for right lung). These findings suggest that during the IME test, EMG activity in the muscles of the lower rib cage increase during progressive respiratory workloads is associated with a greater radioaerosol deposition in the medium third and intermediate and central segments of the lungs. PMID- 16713404 TI - A GC-MS/MS method for the quantitative analysis of low levels of the tyrosine metabolites maleylacetone, succinylacetone, and the tyrosine metabolism inhibitor dichloroacetate in biological fluids and tissues. AB - We developed a sensitive method to quantitate the tyrosine metabolites maleylacetone (MA) and succinylacetone (SA) and the tyrosine metabolism inhibitor dichloroacetate (DCA) in biological specimens. Accumulation of these metabolites may be responsible for the toxicity observed when exposed to DCA. Detection limits of previous methods are 200 ng/mL (1.2 pmol/microL) (MA) and 2.6 microg/mL (16.5 pmol/microL) (SA) but the metabolites are likely present in lower levels in biological specimens. To increase sensitivity, analytes were extracted from liver, urine, plasma and cultured nerve cells before and after dosing with DCA, derivatized to their pentafluorobenzyl esters, and analyzed via GC-MS/MS. PMID- 16713405 TI - Identification of conjugated linoleic acid elongation and beta-oxidation products by coupled silver-ion HPLC APPI-MS. AB - Atmospheric pressure photoionisation (APPI) was used in combination with silver ion (Ag(+))-HPLC for detection of (conjugated) fatty acid methyl esters (FAME) by tandem-mass spectrometry. APPI-MS of methyl esters of conjugated linoleic acid showed an increase in signal-to-noise ratio by a factor of 40 compared to atmospheric pressure chemical ionization in the positive mode. It was possible to identify double bond position, configuration and chain length of FAME based on chromatographic separation and mass detection. The developed LC-MS method is useful for the analysis of CLA elongation and beta-oxidation products, especially with trans,trans-configuration, which are difficult to analyze by conventional GC MS techniques. PMID- 16713406 TI - Development and validation of a quantitative assay for the measurement of two HIV fusion inhibitors, enfuvirtide and tifuvirtide, and one metabolite of enfuvirtide (M-20) in human plasma by liquid chromatography-tandem mass spectrometry. AB - A method for the quantification of two peptide HIV-1 fusion inhibitors (enfuvirtide, T-20 and tifuvirtide, T-1249) and one metabolite of enfuvirtide (M 20) in human plasma has been developed and validated, using liquid chromatography coupled with electrospray tandem mass spectrometry (LC-MS/MS). The analytes were extracted from plasma by solid-phase extraction (SPE) on vinyl-copolymer cartridges. Chromatographic separation of the peptides was performed on a Symmetry 300 C(18) column (50mmx2.1mm I.D., particle size 3.5 microm), using a water-acetonitrile gradient containing 0.25% (v/v) formic acid. The triple quadrupole mass spectrometer was operated in the positive ion-mode and multiple reaction monitoring (MRM) was used for peak detection. Deuterated (d60) enfuvirtide and (d50) tifuvirtide were used as internal standards. The assay was linear over a concentration range of 20-10,000 ng/ml for enfuvirtide and tifuvirtide and of 20-2000 ng/ml for M-20. Intra- and inter-assay precisions and deviations from the nominal concentrations were 0.7, P < 0.001). Serum TGF-beta1 correlated with bone turnover (DPD/Cr, osteocalcin) and was negatively correlated to DBM. Thus, in osteopenic mice, sequential treatment with bFGF followed by risedronate increased trabecular bone microarchitecture, DBM, and bone strength. In addition, suppression of the serum TGF-beta1 with risedronate was associated with increased DBM. Therefore, sequential treatment with bFGF and Ris restores trabecular architecture and allows mineralization of bone to increase, which appears to be beneficial to bone strength. PMID- 16713416 TI - Imaging of the posterolateral corner of the knee. AB - The structures of the posterolateral corner of the knee are increasingly recognized as fulfilling an important role in maintaining knee stability. The posterolateral aspect of the knee is stabilized by a complex anatomy of osseous, myotendinous and ligamentous structures. Unrecognized injuries to this part of the knee are a cause of failure of cruciate ligament reconstruction. This review focuses on the anatomy and common injury patterns involving the posterolateral corner of the knee, with emphasis on magnetic resonance imaging. PMID- 16713417 TI - Imaging of diaphragmatic rupture after trauma. AB - Traumatic rupture of the diaphragm usually results from blunt or penetrating injuries, or iatrogenic causes. Most cases are initially overlooked in the acute phase because they present with variable clinical and radiological signs. An overlooked diaphragmatic injury presents as a hernia many years later with potentially serious complications, therefore selection of the most appropriate radiological technique and accurate diagnosis of traumatic diaphragmatic hernias (DH) on the first admission is important. Although the diagnosis of diaphragmatic injuries is problematic, various investigations may be used for diagnosis. We describe the imaging findings of 19 traumatic DH cases with various imaging techniques. The patients were acute trauma cases or cases with prior trauma or thoraco-abdominal surgery with clinical suspicion of DH. An evaluation of the imaging techniques used in the diagnosis of DH is presented. PMID- 16713418 TI - Two-view mammography at the incident round has improved the rate of screen detected breast cancer in Wales. AB - AIM: To investigate whether pre-invasive and invasive cancer detection rates were improved in Wales after the introduction of two views at incident screens. METHODS: The records of women attending follow-up screening for 2 years before and 2 years after the introduction of two-view incident screening were analysed. Cancer detection rates were compared before and after introduction of two view screening. RESULTS: At the incident round 98,752 women had one and 95,464 had two views. Five hundred and fifty-five cancers were detected with one view and 744 with two, an increased detection rate from 5.6 to 7.8 cancers per 1000 women screened (p=0.01). Two hundred and thirty-nine small cancers were detected with one view and 323 with two, increasing the detection rate from 2.4 to 3.4 per 1000 women screened (p=0.05). CONCLUSIONS: Two-view mammography at incident rounds detects more cancers and more favourable prognosis small cancers than single-view mammograms. PMID- 16713419 TI - CT colonography in a Korean population with a high residue diet: comparison between wet and dry preparations. AB - AIM: To compare wet and dry preparation methods for computed tomography colonography (CTC) in terms of preparation quality, interpretation time, and diagnostic performance for polyp detection in a population with a high residue diet. MATERIALS AND METHODS: Eighty-six patients were divided into two groups. Group 1 (n=24) received a wet preparation of 4l polyethylene glycol (PEG) solution, and group 2 (n=62) received a dry preparation of phosphor-soda. Abnormal findings, including polyps, and the time required to interpret the CTC images in both groups were documented by a radiologist. CTC findings were compared to those of colonoscopy as a reference standard. Two radiologists evaluated the quality of CTC with regard to residual fluid, faeces, and colonic distension using a four-point scale in consensus. Statistical differences for residual fluid, faeces, distensibility on CTC, and interpretation time between the two groups were analysed. The diagnostic performance of CTC in both groups was also compared. RESULTS: One-hundred and ninety polyps in 70 patients were identified using colonoscopy. Regarding the quality of images produced the wet preparation was significantly better than the dry preparation (p<0.05). The average interpretation time was significantly shorter for the wet group (11.7 min) than the dry group (16.4 min) (p<0.05). For per-patient analysis, the positive predictive value (PPV) was significantly better for the wet (100%) than the dry group (79.6%; p=0.025). Sensitivities and PPV for >or=10 mm polyps were comparable between two groups (p>0.05). CONCLUSION: In a population with a high residue diet, CTC with wet preparation can be interpreted in a time-efficient manner and is comparable with CTC with dry preparation. PMID- 16713420 TI - MRI findings and spreading patterns of necrotizing external otitis: is a poor outcome predictable? AB - AIM: To evaluate spreading patterns of necrotizing external otitis (NEO) by magnetic resonance imaging (MRI) and to identify spreading patterns related to a poor outcome. MATERIALS AND METHODS: Fourteen patients with NEO were divided into good and poor outcome groups according to their final clinical outcomes. Initial MRI images were retrospectively reviewed for regional abnormalities, and follow up MRI images were reviewed for ICA flow void abnormality and for the following five spreading patterns: medial, crossed, anterior, intracranial, and combined. The frequencies of the abnormal flow void or spreading patterns were compared between the good and poor response groups. RESULTS: Seven (50%) and seven (50%) patients were respectively allocated to the good and poor outcome groups. Retrocondylar fat infiltration was the most commonest finding on initial MRI images (93%). The frequencies of the abnormal flow void and spreading patterns in the good and poor groups, respectively, were: abnormal flow void, 0 and four (57%); anterior, two (29%) and three (43%); medial, six (86%) and seven (100%); crossed, six (86%) and seven (100%); intracranial middle cranial fossa, one (14%) and four (57%); intracranial posterior cranial fossa, four (57%) and six (86%); intracranial foramen magnum, one (14%) and six (86%). CONCLUSIONS: NEO almost always involves the retrocondylar fat and spreads via various pathways to extracranial or intracranial spaces. The presence of an abnormal flow void and intracranial dural enhancement, particularly in the middle cranial fossa and foramen magnum, may indicate a poor prognosis. PMID- 16713421 TI - Postoperative three-dimensional CT angiography after cerebral aneurysm clipping with titanium clips: detection with single detector CT. Comparison with intra arterial digital subtraction angiography. AB - AIM: To assess the significance of three-dimensional computed tomography angiography (3D-CTA) in detecting remnant necks after cerebral aneurysm clipping. MATERIALS AND METHODS: A total of 59 patients (77 aneurysms) underwent surgery using titanium clips. Two blinded observers independently evaluated the presence of neck remnants on shaded-surface display (SSD) imaging, volume rendered (VR) imaging, and intra-arterial digital subtraction angiography (IADSA). RESULTS: Mean sensitivity and specificity for detecting neck remnants were 50.0 and 74.2% for SSD imaging, 61.5 and 82.8% for VR imaging, and 92.3 and 92.2% for IADSA, respectively. Receiver operating characteristic (ROC) analysis revealed excellent diagnostic performance for IADSA [mean area under ROC curve (Az)=0.97], and good diagnostic performance for 3D-CTA (Az=0.70 and 0.76 for SSD and VR, respectively). Specificity of VR was better than that SSD (p=0.082), however, there was no significant difference between them. CONCLUSION: Use of 3D-CTA techniques can facilitate postoperative evaluation. PMID- 16713422 TI - Prevalence and clinical significance of pleural microbubbles in computed tomography of thoracic empyema. AB - AIM: To determine the prevalence and clinical significance of pleural microbubbles in thoracic empyema. MATERIALS AND METHODS: The charts of 71 consecutive patients with empyema were retrospectively reviewed for relevant demographic, laboratory, microbiological, therapeutic and outcome data. Computed tomography (CT) images were reviewed for various signs of empyema as well as pleural microbubbles. Two patient groups, with and without microbubbles were compared. RESULTS: Mean patient age was 49 years and 72% were males. Microbubbles were detected in 58% of patients. There were no significant differences between patients with and without microbubbles in regard to pleural fluid chemistry. A causative organism was identified in about 75% of cases in both. There was no difference in the rates of pleural thickening and enhancement, increased extra pleural fat attenuation, air-fluid levels or loculations. Microbubbles were diagnosed after a mean of 7.8 days from admission. Thoracentesis before CT was performed in 90 and 57% of patients with and without microbubbles (p=0.0015), respectively. Patients with microbubbles were more likely to require repeated drainage (65.9 versus 36.7%, p=0.015) and surgical decortication (31.7 versus 6.7%, p=0.011). Mortalities were 9.8 and 6.6% respectively (p=0.53). CONCLUSION: Pleural microbubbles are commonly encountered in CT imaging of empyema but have not been systematically studied to date. Microbubbles may be associated with adverse outcome such as repeated drainage or surgical decortication. The sensitivity and specificity of this finding and its prognostic implications need further assessment. PMID- 16713423 TI - Large renal angiomyolipomas: digital subtraction angiographic grading and presentation with bleeding. AB - AIM: To investigate whether a grading system of renal AMLs based on digital subtraction angiography (DSA) and computerized tomography (CT), could help to select patients for embolization. MATERIALS AND METHODS: Thirty patients with 35 renal angiomyolipomas (size range 4-20 cm, mean 9.9 cm) underwent both digital subtraction angiography (DSA) and computed tomography (CT). Based on the DSA appearance the tumours were graded into three grades: grade I, minimal vascularity; grade II, moderate vascularity; grade III, marked vascularity. RESULTS: There were seven grade I (mean 9.3 cm, range 4.5-20 cm), 18 grade II (mean 8.9 cm, range 5-18 cm) and 10 grade III tumours (mean 12.1 cm, range 4-20 cm). Five grade I tumours did not cause symptoms (71%) and two caused flank pain and haematuria, respectively (14.3% each). Nine of the grade II tumours were asymptomatic (50%), seven caused bleeding (39%) and two caused flank pain (11%). Four grade III tumours were asymptomatic (40%), five caused bleeding (50%) and one pain (10%). CONCLUSIONS: According to our criteria, large angiomyolipomas with minimal vascularity are less likely to bleed, and do not need prophylactic treatment. This needs to be confirmed in larger studies. PMID- 16713424 TI - Case report: recurrent posterior sacculation of the pregnant uterus and placenta increta. PMID- 16713425 TI - Case report: ureteroiliac fistula: a late sequela of radiotherapy and long-term ureteric stent placement. PMID- 16713426 TI - Distribution and metabolic syndrome correlates of plasma C-reactive protein in biracial (black-white) younger adults: the Bogalusa Heart Study. AB - The association between plasma C-reactive protein (CRP), a marker of systemic inflammation, and the metabolic syndrome is well recognized. However, information is scant regarding the component of metabolic syndrome that is critical in modulating CRP levels in younger adults. This aspect was examined in a biracial (black-white) community-based sample of 1083 younger adults (mean age, 36.1 years; 71% white, 45% male) enrolled in the Bogalusa Heart Study. Plasma CRP along with metabolic syndrome variables were measured. CRP levels showed a significant race (black>white, P=.01) and sex (female>male, P=.0001) differences, and related to measures of obesity (body mass index [BMI], waist circumference, and sagittal diameter), blood pressure (systolic, diastolic, and mean arterial blood pressure), lipoproteins (low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, triglycerides, total cholesterol-high-density lipoprotein cholesterol ratio), glucose homeostasis (insulin, glucose, homeostasis model assessment of insulin resistance), and uric acid, after adjusting for age, race, sex, and cigarette smoking. Furthermore, CRP levels increased with increasing number of metabolic syndrome risk factors, as defined by the National Cholesterol Education Program Adult Treatment Panel III, regardless of race and sex (P for trend<0001). In multivariate analysis, using 3 separate models for different obesity measures, obesity was the major contributor to the explained variance in each model with BMI, waist, and sagittal diameter contributing 17.0%, 13.4%, and 17.1% of the variance, respectively. In contrast, homeostasis model assessment of insulin resistance contributed 1.2%, 0.3%, and 0% to the explained variance in the models with BMI, waist, and sagittal diameter, respectively. In conclusion, CRP levels differ among race and sex groups and correlate to metabolic syndrome variables in younger adults. In addition, these findings strongly suggest that although obesity and insulin resistance are the main underlying features of the metabolic syndrome, the former appears to be the major mediator of CRP levels, which has important health implications. PMID- 16713427 TI - Lipid and lipoprotein profiles in Ethiopian patients with diabetes mellitus. AB - The association between dyslipidemia and diabetes mellitus is well established. Although various lipoprotein abnormalities have been described in patients with diabetes mellitus elsewhere, there is limited information from African patients. We undertook a cross-sectional study to assess the prevalence of dyslipidemia in Ethiopian patients with types 1 and 2 diabetes. A total of 193 subjects were included in the study (54 patients had type 1 diabetes mellitus, 92 patients had type 2 diabetes mellitus, and 47 were nondiabetic controls). Of these, 93 (48.6%) were men and 103 (51.4%) were women. The mean age+/-SEM for patients with type 1 diabetes mellitus, type 2 diabetes mellitus, and controls were 29.8+/-1.4, 51.2+/ 1.1, and 29.0+/-1.7 years, respectively. Hypercholesterolemia and hypertriglyceridemia, defined as cholesterol level of greater than 5.2 mmol/L and triglyceride level of greater than 1.8 mmol/L, were found in 47.3% and 41.8% of patients with diabetes mellitus compared with 27% and 17% in controls (P<.05 for both). The mean total cholesterol level+/-SEM was significantly higher in patients with type 1 or 2 diabetes mellitus than controls (5.76+/-0.27 mmol/L in type 1 diabetes mellitus, 5.25+/-0.2 mmol/L in type 2 diabetes mellitus, and 4.67+/-0.28 mmol/L in healthy controls, P<.02). Triglycerides and low-density lipoprotein levels were also significantly higher in patients with diabetes than in controls, whereas high-density lipoprotein levels were significantly lower in patients with diabetes. In conclusion, our study demonstrates that in Ethiopians with diabetes mellitus, dyslipidemia occurs more frequently than in controls. Thus, we recommend periodic screening for dyslipidemia in all Ethiopian patients with diabetes. Other studies are needed to assess the potential negative effect of dyslipidemia and obesity on morbidity and mortality in Ethiopians with diabetes. PMID- 16713428 TI - N-Glucuronidation of the antiepileptic drug retigabine: results from studies with human volunteers, heterologously expressed human UGTs, human liver, kidney, and liver microsomal membranes of Crigler-Najjar type II. AB - Retigabine (D-23129), an N-2-amino-4-(4-fluorobenzylamino)phenylcarbamine acid ethyl ester, is a novel antiepileptic drug which is currently in phase II clinical development. This drug undergoes N-glucuronidation. We aimed to identify the principal enzymes involved in the N-glucuronidation pathway of retigabine and compared our findings with those obtained from human liver (a pool of 30 donors) and kidney microsomes (a pool of 3 donors) and with results from a human absorption, distribution, metabolism, and excretion study upon administration of 200 microCi of [(14)C]-D-23129. Essentially, microsomal assays with UGT1A1 produced only one of the 2 N-glucuronides, whereas UGT1A9 is capable of forming both N-glucuronides. The rates of metabolism for UGT1A9, human liver microsomes, and UGT1A1 were 200, 100, and 100 pmol N-glucuronide per minute per milligram of protein, respectively. At the 50 micromol/L uridine diphosphate glucoronic acid (UDPGA) concentration, UGT1A4 also catalyzed the N-glucuronidation of retigabine, the rates being approximately 5 and 6 pmol/(min.mg protein). With UGT1A9, the production of metabolites 1 and 2 proceeded at a K(m) of 38+/-25 and 45+/-15 micromol/L, whereas the K(m) for retigabine N-glucuronidation by human liver microsomal fractions was 145+/-39 micromol/L. Furthermore, a V(max) of 1.2+/-0.3 (nmol/[min.mg protein]) was estimated for human liver microsomes (4 individual donors). We investigated the potential for drug-drug interaction using the antiepileptic drugs valproic acid, lamotrigine, the tricyclic antidepressant imipramine, and the anesthetic propofol. These are commonly used medications and are extensively glucuronidated. No potential for drug-drug interactions was found at clinically relevant concentrations (when assayed with human liver microsomes or UGT1A9 enzyme preparations). Notably, the biosynthesis of retigabine-N glucuronides was not inhibited in human liver microsomal assays in the presence of 330 micromol/L bilirubin, and glucuronidation of retigabine was also observed with microsomal preparations from human kidney and Crigler-Najjar type II liver. This suggests that lack of a particular UDP-glucuronosyltransferase (UGT) isoform (eg, UGT1A1 in kidney) or functional loss of an entire UGT1A gene does not completely abolish disposal of the drug. Finally, chromatographic separations of extracts from microsomal assays and human urine of volunteers receiving a single dose of (14)C-retigabine provided clear evidence for the presence of the 2 N glucuronides known to be produced by UGT1A9. We therefore suggest N glucuronidation of retigabine to be of importance in the metabolic clearance of this drug. PMID- 16713429 TI - Gliclazide protects 3T3L1 adipocytes against insulin resistance induced by hydrogen peroxide with restoration of GLUT4 translocation. AB - Increased oxidative stress under hyperglycemia may contribute to progressive deterioration of peripheral insulin sensitivity. In this study, we investigated whether gliclazide, a second-generation sulfonylurea, can protect 3T3L1 adipocytes from insulin resistance induced by oxidative stress, and whether gliclazide can restore insulin-stimulated glucose transporter 4 (GLUT4) translocation under oxidative stress. We incubated 3T3L1 adipocytes in hydrogen peroxide to produce oxidative stress, then administered various concentrations of gliclazide, N-acetylcystein (NAC), or glibenclamide. Cells treated with these drugs were next exposed to insulin, subsequent glucose uptake was measured, and the insulin-stimulated GLUT4 translocation was monitored in living cells. We found that hydrogen peroxide treatment alone suppressed glucose uptake by insulin stimulation to 65.9%+/-7.8% of the corresponding controls (P<.01). However, addition of 0.1 to 10 micromol/L gliclazide to hydrogen peroxide-treated cells dose-dependently restored glucose uptake, with 5 micromol/L gliclazide significantly restoring glucose uptake to 93.3+/-6.6% (P<.01) even under hydrogen peroxide. Treatment with the known anti-oxidant NAC also dose-dependently (0.1-10 mmol/L) restored insulin-induced glucose uptake in the presence of hydrogen peroxide. However, glibenclamide (0.1-10 micromol/L), another second-generation sulfonylurea, failed to improve glucose uptake. Similarly, treatment with 5 micromol/L gliclazide or 10 mmol/L NAC significantly overcome the reduction in insulin-stimulated GLUT4 translocation by hydrogen peroxide (P<.01), whereas 5 micromol/L glibenclamide did not. Therefore our data regarding gliclazide further characterize its mechanism of hypoglycemic effect: the observed improvements in insulin sensitivity and in GLUT4 translocation indicate that gliclazide counters the hydrogen peroxide-induced insulin resistance in 3T3L1 adipocytes and also would further augment the hypoglycemic effect of this drug as insulinotropic sulfonylurea. PMID- 16713430 TI - Effects of fenofibrate on lipid metabolism in adipose tissue of rats. AB - The effect of fenofibrate, a peroxisome proliferator-activated receptor alpha agonist, on body weight gain and on reduction of adipose tissue pads has been ascribed to increased fat catabolism in liver mainly through the induction of target enzymes involved in hepatic lipid metabolism. The aim of this study was to investigate whether peroxisome proliferator-activated receptor alpha activation also affects metabolic pathways in adipose tissue of rats treated with fenofibrate (100 mg/kg body weight) for 9 days. Fenofibrate lowered body weight gain and plasma triglyceride, total cholesterol, and high-density lipoprotein cholesterol but had no influence on food intake and on plasma glucose levels. The activity of lipoprotein lipase of treated animals decreased 50% in epididymal, 29% in retroperitoneal, and was not affected in the mesenteric fat pads. In this study, we show a 34% decrease in epididymal adipose tissue de novo lipogenesis by fenofibrate. The results demonstrate that insulin sensitivity of lipolysis is decreased in fenofibrate-treated rats which resulted in 30% higher rate of glycerol release when compared to the control group. These findings suggest that besides its effects on liver, fenofibrate exerts effects on lipid metabolism in adipose tissue which may contribute to decreasing adiposity. PMID- 16713431 TI - Active acromegaly enhances spontaneous parathyroid hormone pulsatility. AB - In healthy subjects, parathyroid hormone (PTH) is secreted in a dual fashion, with low-amplitude and high-frequency pulses superimposed on tonic secretion. These 2 components of PTH secretion seem to have different effects on target organs. The aim of our study was to evaluate whether growth hormone excess in acromegaly may modify the spontaneous pulsatility of PTH. Five male patients with newly diagnosed active acromegaly and 8 healthy subjects were evaluated by 3 minute blood sampling for 6 hours. Plasma PTH concentrations were evaluated by multiparameter deconvolution analysis. Plasma PTH release profiles were also subjected to an approximate entropy (ApEn) estimate, which provides an ensemble measure of the serial regularity or orderliness of the release process. In acromegalic patients, baseline serum PTH values were not significantly different from those measured in the healthy subjects, as well as tonic PTH secretion rate, number of bursts, fractional pulsatile PTH secretion, and ApEn ratio. Conversely, PTH pulse half-duration was significantly longer in acromegalic patients vs healthy subjects (11.8+/-0.95 vs 6.9+/-1.6 minutes; P=.05), whereas PTH pulse mass showed a tendency (P=.06) to be significantly greater in acromegalic patients. These preliminary data suggest that growth hormone excess may affect PTH secretory dynamics in patients with acromegaly. Potentially negative bone effects of the modifications of PTH secretory pattern in acromegaly should be investigated. PMID- 16713432 TI - Effects of alendronate combined with hormone replacement therapy on osteoporotic postmenopausal Chinese women. AB - To evaluate the effect of alendronate combined with hormone replacement therapy (HRT) on postmenopausal osteoporotic Chinese women living in Taiwan, we treated 151 women (age range, 47-70 years; mean, 61 years) with conjugated equine estrogen (0.625 mg), medroxyprogesterone 5 mg, and elemental calcium 500 mg daily with either alendronate 10 mg (n = 79) or placebo (n = 72), and measured their bone mineral density (BMD) at the lumbar spine and hip every 6 months for 3 years. Urine N-telopeptide of type I collagen corrected by concentration of urine creatinine (NTx/Cr) and serum osteocalcin (OC) concentration was also measured at weeks 2, 4, and every 3 months from month 3 for 2 years. Significantly higher percentage increases in BMD at the lumbar spine (P < .0001, 2-way analysis of variance) throughout the 36-month treatment period were found in the alendronate plus HRT group than in the HRT-only group. However, there was no difference in BMD at the femoral neck and trochanter between these 2 groups. Treatment with alendronate plus HRT resulted in a 10.1% increase at the L-spine BMD and a 7.7% increase at the trochanter BMD at the end of the 3-year study period (P < .01, compared with baseline at both sites). A significant decline in urine NTx/Cr was observed at week 4 in the alendronate plus HRT group, whereas in the HRT-only group, a significant decline in urine NTx/Cr occurred at month 9. By the end of 24 months, urine NTx/Cr decreased by 49.7% in the alendronate plus HRT group (P = .001 compared with a 20.4% increase in the HRT group). A significant decline in serum OC level occurred at month 3 in the alendronate plus HRT group, whereas a similar decline was observed at month 6 in the HRT-only group. By the end of 24 months, serum OC decreased by 52.2% in the alendronate plus HRT group (P < .001 compared with a 1.5% increase in the HRT-only group). Subjects treated with alendronate plus HRT had a significantly greater percentage decrease in urine NTx/Cr (P = .0001) and serum OC (P = .0007) than subjects treated with HRT only throughout the 24-month treatment period by 2-way analysis of variance comparison. There was no difference in upper gastrointestinal or drug-related side effects between groups. In conclusion, our data suggest that the use of alendronate combined with HRT for 3 years was well tolerated and it significantly increased BMD at the L-spine and hip in postmenopausal Chinese women with osteoporosis. This regimen is safe and can be used in subjects who have no satisfactory response to a single agent or who have very low BMD with multiple risks. However, this study does not indicate whether HRT plus alendronate has any greater effect on BMD than alendronate alone. PMID- 16713433 TI - Role of thyrotropin in metabolism of thyroid hormones in nonthyroidal tissues. AB - T(4) conversion into T(3) in peripheral tissues is the major source of circulating T(3). However, the exact mechanism of this process is ill defined. Several in vitro studies have demonstrated that thyrotropin facilitates deiodination of T(4) into T(3) in liver and kidneys. However, there is a paucity of in vitro studies confirming this activity of thyrotropin. Therefore, this study was conducted to examine the influence of thyrotropin on thyroid hormone metabolism in nonthyroidal tissues. We assessed T(4), T(3), reverse T(3) (rT(3)), and T(3) resin uptake (T(3)RU) responses up to 12 hours at intervals of 4 hours in 6 thyroidectomized female mongrel dogs rendered euthyroid with LT(4) replacement therapy before and after subcutaneous (SC) administration of bovine thyrotropin (5 U) on one day and normal saline (0.5 mL) on another in a randomized sequence between 08:00 and 09:00 am. Euthyroid state after LT(4) replacement was confirmed before thyrotropin administration. Serum T(4), T(3), rT(3), and T(3)RU all remained unaltered after SC administration of normal saline. No significant alteration was noted in serum T(3)RU values on SC administration of thyrotropin. However, serum T(3) rose progressively reaching a peak at 12 hours with simultaneous declines being noted in both serum T(4) and rT(3) concentrations (P < .05 vs prethyrotropin values for all determinations). The changes after SC administration were significantly different (P < .001) in comparison to those noted on SC administration of normal saline. Thyrotropin may promote both the conversion of T(4) to T(3) and metabolism of rT(3) into T(2) in nonthyroidal tissues via enhancement of the same monodeionase. PMID- 16713434 TI - G-protein beta 3 subunit polymorphism C1429T and low-density lipoprotein receptor related protein 5 polymorphism A1330V are risk factors for hypercholesterolemia in Japanese males--a prospective study over 5 years. AB - We examined the relationship between the C825T, C1429T, and A-350G variants in the G-protein beta 3 subunit (GNB3) gene, the A1330V and Q89R variants in the low density lipoprotein receptor-related protein 5 (LRP5) gene, and the risk of hypercholesterolemia in a prospective study in Japanese workers. This study included observations over a 5-year period from 1997 to 2002 on 936 males and 662 females who were not hypercholesterolemic on entry. Hypercholesterolemia was defined as a serum total cholesterol level of 240 mg/dL or higher. Pooled logistic regression analyses were performed using either of the gene variants with age, body mass index, smoking, alcohol consumption, and habitual exercise as the covariates. The risk of the development of hypercholesterolemia was 2.27 times higher in males with the TT genotype of GNB3/C1429T than in males with the CC genotype (95% confidence interval, 1.04-4.94), after adjustment for the effects of other potential covariates. Simultaneously, the risk was 1.49 times higher in males with the AV genotype of LRP5/A1330V than in males with the AA genotype (95% confidence interval, 1.04-2.12) after adjustment for the effects of other potential covariates. This study indicates the GNB3/C1429T and LRP5/A1330V are independent risk factors for hypercholesterolemia in Japanese males and suggests that targeting these polymorphisms may be beneficial when attempting to prevent hypercholesterolemia in the general Japanese male population. PMID- 16713435 TI - The effect of high-dose simvastatin on free fatty acid metabolism in patients with type 2 diabetes mellitus. AB - Statins improve all major lipid fractions, reduce coronary heart disease risk, and may have a minor effect on glucose tolerance. A reduction in free fatty acid flux and concentrations could be partly responsible for these effects. We measured nocturnal and postprandial plasma palmitate concentrations and rate of appearance (R(a)) on 2 occasions in 12 obese dyslipidemic subjects with type 2 diabetes mellitus, using a single-blind, crossover format (placebo followed by simvastatin, 80 mg/d), and also on 1 occasion in 6 untreated control subjects. The diabetic subjects had increased average nocturnal (127+/-13 vs 80+/-10 micromol/L, P<.05) and 2-hour postprandial (49+/-6 vs 17+/-2 micromol/L, P<.001) palmitate concentrations, as well as increased nocturnal (31.6+/-3.7 vs 19.5+/ 3.7 mmol/m(2) over 9 hours, P<.05) and postprandial (11.5+/-3.7 vs 5.5+/-3.7 mmol/m(2) every 4 hours, P<.005) integrated palmitate R(a) compared to normal controls. High-dose simvastatin reduced serum triglycerides by 35% but had no effect on plasma palmitate concentrations or R(a). These results suggest that the triglyceride-lowering effect of statins is not mediated through an effect on FFA metabolism. PMID- 16713437 TI - Asymmetric dimethylarginine and hemodynamic regulation in middle-aged men. AB - The goal of this study was to evaluate the role of asymmetric dimethylarginine (ADMA) in the regulation of hemodynamic functions in hypertensive men. It has been suggested that ADMA, as an endogenous nitric oxide synthase inhibitor, is linked to hypertension and vascular reactivity. Sixty-seven men aged 51.1 years (range, 45-55 years) were studied. Plasma ADMA and symmetric dimethylarginine were determined by high-performance liquid chromatography-tandem mass spectrometry. Blood pressure (BP) was measured by 24-hour ambulatory recordings and casual measurements. Hemodynamic regulation was assessed by noninvasive methods. The nitric oxide production was estimated based on plasma nitrate (NO(3)(-)) determination. Results showed that plasma arginine derivatives or l arginine/ADMA ratio was not associated with BP values observed during 24-hour monitoring or in casual measurements. Systemic vascular resistance, pulse wave velocity, or cardiac output was not associated with plasma ADMA or plasma NO(3)( ) levels. No association was found between plasma ADMA and NO(3)(-) either. Interestingly, subjects on antihypertensive treatment had lower plasma ADMA concentrations than nontreated subjects (0.30+/-0.08 and 0.36+/-0.11 micromol/L, respectively, P=.04) despite higher BP values. In conclusion, these results suggest that plasma ADMA does not have a determinative role in the regulation of hemodynamic functions in Finnish middle-aged men. PMID- 16713436 TI - An imbalance in the methionine content of the maternal diet reduces postnatal growth in the rat. AB - The pregnant rat fed a low-protein diet has become widely used as a model system in the study of the prenatal programming of adult metabolism and disease. When pregnant rats of the hooded Lister strain were fed semisynthetic diets containing 18% or 9% casein supplemented with 0.5% dl-methionine, there was significant postnatal mortality in the group fed the low-protein diet. In a second experiment, dams were fed diets containing 9% casein supplemented with varying concentrations of dl-methionine up to 0.4% (w/w) and compared with a group fed a diet containing 18% casein supplemented with 0.5% dl-methionine. At birth, the pups from dams fed the low-protein diets supplemented with 0.2% dl-methionine or greater were significantly smaller than those of the dams fed the diet containing 18% protein. By 25 weeks of age, the body weight of the offspring of dams fed the low-protein diet supplemented with 0.2% or 0.3% dl-methionine were approximately 10% lower than those in the control group of offspring from dams fed 18% protein supplemented with 0.5% dl-methionine. There were corresponding changes in the weights of the major organs. These data suggest that increasing the dl-methionine supplement in the low-protein diet retards the growth of the fetus and affects the mature adult body weight. In contrast to the findings of other studies that used different formulas of the low-protein diet, the glucose tolerance in the offspring was unaffected by the protein content of the maternal diet at all levels of dl-methionine supplementation. These results suggest that the changes in metabolism of the offspring result from interactions between protein, lipids, and carbohydrates in the maternal diet, rather than a consequence of postnatal growth retardation per se and highlight the importance of considering all components of the maternal diet in the programming mechanism. PMID- 16713438 TI - Gliclazide inhibits differentiation-associated biologic events in human monocyte derived macrophages. AB - We investigated the in vitro effect of gliclazide on human monocyte-derived macrophage scavenger receptor expression and activity, foam cell formation, and lipopolysaccharide-induced cytokine production. Differentiation of human monocytes into macrophages in the presence of gliclazide (1-10 microg/mL) decreased CD36 expression by 20% to 50%, with maximal effect occurring at 2.5 microg/mL (P<.05). This effect was mimicked by vitamin E (50 micromol/L) and N acetyl-L-cysteine (10 mmol/L). Incubation of the cells with gliclazide and N acetyl-L-cysteine also reduced CD36 activity by 30% (P<.02). Despite these effects, neither gliclazide nor vitamin E did affect foam cell formation. In contrast, gliclazide significantly reduced lipopolysaccharide-stimulated macrophage tumor necrosis factor alpha and interleukin 6 secretion (P<.05). Overall, these data indicate that gliclazide, at concentrations in the therapeutic range, may regulate some key biologic events associated with the process of monocyte differentiation into macrophages. PMID- 16713439 TI - An alpha-glucosidase inhibitor, voglibose, reduces oxidative stress markers and soluble intercellular adhesion molecule 1 in obese type 2 diabetic patients. AB - Postprandial hyperglycemia and hyperlipidemia are considered risk factors for cardiovascular disease. This study was designed to elucidate whether improving the postprandial state by voglibose, an alpha-glucosidase inhibitor, leads to the reduction of oxidative stress markers and soluble adhesion molecules in obese type 2 diabetic patients. A total of 30 Japanese obese type 2 diabetic patients were randomly assigned and treated for 3 weeks with either diet alone (the control group) or diet plus voglibose (0.9 mg daily) (the voglibose group) (n=15 each). Analysis of the diurnal metabolic profiles revealed a significant reduction of postprandial hyperglycemia and hyperlipidemia in the voglibose group relative to the control group (P<.05), despite the similar improvement in body mass index and hemoglobin A(1c) in both groups. Voglibose also decreased significantly the plasma levels of soluble intercellular adhesion molecule 1 and urinary excretion of 8-iso-prostaglandin F(2)alpha and 8-hydroxydeoxyguanosine (P<.01) and C-reactive protein (P<.05) relative to the control group. In conclusion, this study represents the first demonstration that voglibose reduces oxidative stress generation and soluble intercellular adhesion molecule 1 in parallel with the reduction of postprandial hyperglycemia and hyperlipidemia in obese type 2 diabetic patients. PMID- 16713440 TI - Rapamycin, an mTOR inhibitor, disrupts triglyceride metabolism in guinea pigs. AB - This study was designed to define some of the mechanisms by which rapamycin (RAPA), an mTOR inhibitor, induces hypertriglyceridemia when used as an immunosuppressive or antiproliferative agent and to determine whether low doses result in less undesirable side effects. Thirty male guinea pigs (n=10 per group) were randomly assigned to control (no RAPA), low-RAPA (0.08 mg/d), or high-RAPA (0.85 mg/d) treatment for 3 weeks. Rapamycin treatment resulted in more than a 2 fold increase in plasma triglycerides (TG) (P<.01), whereas no differences were observed in plasma cholesterol between RAPA and control groups. Low-RAPA treatment resulted in lower concentrations of cholesterol in the aorta (28.6%) and lower hepatic acyl-CoA cholesteryl acyltransferase activity compared to control and high-RAPA groups (P<.01). In addition, acyl-CoA cholesteryl acyltransferase activity was positively correlated with aortic cholesterol (r=0.43, P<.05). In contrast, aortic TG concentrations were higher in RAPA treated guinea pigs than in control (P<.01). Very low density lipoprotein and low density lipoprotein particles isolated from guinea pigs treated with RAPA were larger in size and contained more TG molecules than particles from control animals. Interestingly, plasma free fatty acids and fasting plasma glucose were 65% and 72% higher in the high-RAPA group than in control (P<.01). Tumor necrosis factor-alpha concentrations in the aorta were 3.6- and 10.4-fold higher in the low-RAPA and high-RAPA groups than in control guinea pigs (P<.01). These results suggest that RAPA interferes with TG metabolism by altering the insulin signaling pathway, inducing increased secretion of very low density lipoprotein and promoting deposition of TG in the aorta. Low RAPA was found to decrease cholesterol accumulation in tissue (liver and aorta) compared to high RAPA, suggesting that lower doses could be less detrimental to transplant patients. PMID- 16713441 TI - Chronic ethanol consumption upregulates the cytosolic and plasma membrane sialidase genes, but down regulates lysosomal membrane sialidase gene in rat liver. AB - We have previously shown that chronic ethanol feeding stimulates liver cytosolic sialidase (CS) and plasma membrane sialidase (PMS), whereas it decreases lysosomal membrane sialidase (LMS) activities with concomitant alterations in their relative synthetic rate in rat. To understand the molecular mechanism(s) for these changes, we have evaluated the effect of ethanol administration in male Wistar rats as a function of increasing dietary ethanol concentration after 8 weeks of pair-feeding on (i) the expression of CS, PMS, and LMS genes by real time quantitative polymerase chain reaction method; (ii) their relative transcription rates by nuclear run-on assay; and (iii) the actual amount of these sialidase proteins in the liver fractions of the respective groups by Western blot method. We have demonstrated that the animals fed with 10.6%, 20.8%, and 36% of total calories as ethanol showed a 20% (P<.05), 34% (P<.01), and 69% (P<.01) increase in CS mRNA level, and 22% (P<.05), 26% (P<.01), and 47% (P<.01) increase in PMS mRNA level, but a decrease in LMS mRNA level by 35% (P<.05), 50% (P<.01), and 80% (P<.01), respectively, as compared to controls. Western blot analyses of CS, PMS, and LMS in the liver subfractions showed that changes in protein levels of CS, LMS, and PMS were consistent with the corresponding changes in the respective mRNA levels. Thus, the upregulation of CS and PMS, but not LMS which is down regulated by chronic ethanol, may account for the appearance of asialoconjugates in alcoholics. PMID- 16713442 TI - Effect of acute hyperglycemia and/or hyperinsulinemia on polymorphonuclear functions in healthy subjects. AB - Abnormal immune functions of polymorphonuclear (PMN) cells occur in a variety of pathophysiological conditions. There exists a close link between glucose metabolism and PMN functions. The aim of this study was to assess the effect of short-term hyperglycemia and/or hyperinsulinemia on phagocytosis and respiratory burst of PMN cells in healthy subjects in vivo. The study was performed on 12 healthy subjects (mean age, 26.9+/-1.6 years; body mass index, 24.4+/-0.84 kg/m(2)). Acute hyperglycemia and/or hyperinsulinemia was induced by three 4-hour long clamp studies-hyperglycemic hyperinsulinemic clamp (HHC), hyperinsulinemic euglycemic clamp (HEC), and isolated hyperglycemic clamp with insulin secretion blockade (HGC). Polymorphonuclear cell phagocytosis and PMN cell respiratory burst (mean percentage and mean fluorescent intensity of phagocyting/activated PMN cells, phagocytic, and respiratory burst indexes) were evaluated by flow cytometry under basal and stimulated conditions. Results detected during clamp studies were compared with those found during a control study with saline infusion. Significant reductions in the mean percentage of phagocyting cells measured under basal conditions after the HHC (6.7%+/-1.3% vs 12.1%+/-4.3%; P<.05) and HGC (4.5% +/-1.8% vs 9.9%+/-2.1%; P<.05) were found in comparison with the pre-clamp study period; however, these results did not differ significantly from those detected during the control clamp (CC) study. Significantly higher phagocytic (115.1+/-65 vs 35.8 +/-18.6; P<.05) and respiratory burst indexes (16.5+/-3 vs 10.1+/-1.4; P<.05) measured under basal conditions were found after HEC in comparison with the pre-clamp data. However, these data did not differ significantly from those found after the CC study. No significant differences in other parameters of detected PMN cell immune functions were found after HHC, HEC, and HGC. In conclusion, immune functions of PMN cells were not significantly influenced by short-lasting hyperglycemia and/or hyperinsulinemia induced in vivo by clamp techniques in healthy subjects compared to changes induced by the CC study. Further studies on the short-term effect of glucose metabolism on PMN functions in diabetic patients should be considered necessary. PMID- 16713443 TI - Angiotensinogen gene polymorphism (Met235Thr) influences visceral obesity and insulin resistance in obese Japanese women. AB - To investigate the relationship between angiotensinogen (AGT) Met235Thr polymorphism (M235T) and human obesity, because AGT is regarded as one of the cytokines produced from adipocytes and serum AGT concentrations are reported to be positively correlated with body mass index. One hundred and twenty obese Japanese women (age, 58.8+/-9.4 years; body mass index, 32.2+/-4.9 kg/m(2)) were enrolled. Angiotensinogen genotypes were determined with a fluorescent allele specific DNA primer assay system. Subjects were divided into M/M, M/T, and T/T groups. Control subjects comprised 146 healthy age-matched women. Clinical characteristics and the effects of diet and exercise therapy for 6 months were compared among the 3 genotypes. The genotype frequencies of AGT M235T polymorphism were in accordance with the Hardy-Weinberg equation (obese: M/M, 6.7%; M/T, 27.5%; T/T, 65.8%; control: M/M, 6.8%; M/T, 21.2%; T/T, 71.9%). The frequency of the T allele did not differ between obese and control subjects (0.80 vs 0.83). As the number of obese women with M/M genotype was only 8, comparisons of the characteristics and outcomes of weight reduction therapy were performed only between subjects with M/T genotype and T/T genotype. In the T/T group, % body fat and waist circumference at baseline were significantly greater than in the M/T group (36.3%+/-4.8% vs 33.8%+/-4.7%, P=.0105; 107.9+/-10.9 vs 102.6+/-7.9 cm, P=.0428, respectively). Before the weight reduction therapy, significantly higher insulin and higher homeostasis model assessment (HOMA-R) were demonstrated in the T/T group than in the M/T group (9.1+/-5.5 microU/mL vs 5.9+/-4.4 microU/mL, P=.0056; 2.3+/-1.4 vs 1.6+/-1.3, P=.0252, respectively). Both systolic and diastolic blood pressure at baseline in the T/T group tended to be higher than those in the M/T group, but the differences were not significant. No genotype-dependent difference in energy expenditure or outcome of weight reduction therapy was observed with respect to AGT M235T polymorphism. After the diet and exercise therapy, the blood pressure in the T/T group tended to be higher than that in the M/T group, but the difference was not significant. We demonstrated that the T/T genotype of the AGT M235T gene polymorphism was positively related to visceral obesity and hyperinsulinemia in obese Japanese women. Blood pressure did not show genotype-specific differences before or after the treatment. Further studies of the association between obesity and this gene polymorphism should contribute to understanding and treating obesity-related diseases. PMID- 16713445 TI - alpha-Glucosidase inhibitors prevent diet-induced increases in intestinal sugar transport in diabetic mice. AB - The recommended diet for diabetes mellitus is rich in complex carbohydrates. We have previously shown that high carbohydrate levels in the intestinal lumen induce adaptive increases in sugar absorption which in turn exacerbate postprandial hyperglycemia in diabetic mice. alpha-Glucosidase inhibitors (AGIs) hinder digestion of complex carbohydrates and therefore alleviate postprandial glycemic excursions. In this study, we tested the hypothesis that AGIs prevent the carbohydrate-induced upregulation of intestinal glucose and fructose transport in diabetes. Streptozotocin-diabetic mice were fed the following isocaloric diets: high carbohydrate (H), H plus acarbose (HA), H plus deoxy nojirimycin (HD), and low carbohydrate (L), then nutrient uptakes were determined after 2 and 4 weeks. Body weight, intestinal weight, and length were independent of diet. Fasting and postprandial blood glucose levels were lower in HA and HD than in H mice. Uptakes of D-glucose and D-fructose were 2 to 3 times greater in H than in L mice, but HA and HM diets gradually reduced D-glucose uptakes to rates similar to L mice. Only HA diets reduced D-fructose uptake. Intestinal proline, aspartate, and glutamine uptakes were each greater in L than in H, HA, and HD mice. alpha-Glucosidase inhibitors did not alter intestinal permeability and amino acid transport rates. alpha-Glucosidase inhibitor-inhibitable increases in total intestinal absorptive capacity for sugars were due to carbohydrate induced increases in V(max) of glucose transport. Clearly, one potential mechanism by which AGIs blunt postprandial glycemic excursions and lower fasting blood glucose concentrations in individuals consuming carbohydrate-containing diets is by preventing carbohydrate-induced increases in intestinal sugar transport. PMID- 16713444 TI - Effects of lifestyle modifications on C-reactive protein: contribution of weight loss and improved aerobic capacity. AB - High-sensitivity C-reactive protein (hs-CRP) is associated with an increased risk of cardiovascular disease and the development of type 2 diabetes mellitus. We analyzed the effects of lifestyle modifications including exercise training on hs CRP in 47 overweight and obese adults. Subjects were divided into a lifestyle modification group (n=23) (exercise and diet instruction) and a control group (n=24) who did not participate in any lifestyle modification. After 3 months, body weight (80.8+/-11.5 to 73.5+/-10.7 kg, P<.01), total cholesterol (217+/-38.4 to 178.0+/-25.6 mg/dL, P<.01), low-density lipoprotein cholesterol (151.3+/-34.9 to 116.7+/-27.8 mg/dL, P<.01), Vo(2)peak (30.3+/-5.1 to 37.1+/-6.9 mL/[kg . min], P<.01), and log hs-CRP (0.75+/-0.4 to 0.56+/-0.3 mg/dL, P=.01) were significantly improved in the lifestyle modification group, but there was no significant improvement in the control group. Changes in log hs-CRP were associated with changes in Vo(2)peak (r=-0.41, P=.004) and changes in weight loss (r=0.42, P=.004). In stepwise multiple regression analysis, weight loss (P=.034) and improved Vo(2)peak (P=.039) were independent predictors of the changes in hs-CRP. When grouped into quartiles according to decreasing weight and increasing Vo(2)peak, levels of changes in log hs-CRP improved across quartiles of weight loss (P<.05) and improved Vo(2)peak (P<.01). Thus, lifestyle changes including regular exercise training in overweight and obese adults decreased hs-CRP, and this was associated with weight loss and improved Vo(2)peak. PMID- 16713446 TI - The Src family kinase, Lyn, is activated in pancreatic acinar cells by gastrointestinal hormones/neurotransmitters and growth factors which stimulate its association with numerous other signaling molecules. AB - Src family kinases (SFK) play a central signaling role for growth factors, cytokines, G-protein-coupled receptors and other stimuli. SFKs play important roles in pancreatic acinar cell secretion, endocytosis, growth, cytoskeletal integrity and apoptosis, although little is known of the specific SFKs involved. In this study we demonstrate the SFK, Lyn, is present in rat pancreatic acini and investigate its activation/signaling. Ca(2+)-mobilizing agents, cAMP-mobilizing agents and pancreatic growth factors activated Lyn. CCK, a physiological regulator of pancreatic function, rapidly activated Lyn. The specific SFK inhibitor, PP2, decreased Lyn activation; however, the inactive analogue, PP3, had no effect. Inhibition of CCK-stimulated changes in [Ca(2+)](i) decreased Lyn activation by 55%; GFX, a PKC inhibitor by 36%; and the combination by 95%. CCK activation of Lyn required stimulation of high and low affinity CCK(A) receptor states. CCK stimulated an association of Lyn with PKC-delta, Shc, p125(FAK) and PYK2 as well as with their autophosphorylated forms, but not with Cbl, p85, p130(CAS) or ERK 1/2. These results show Lyn is activated by diverse pancreatic stimulants. CCK's activation of Lyn is likely an important mediator of its ability to cause tyrosine phosphorylation of numerous important cellular mediators such as p125(FAK), PYK2, PKC-delta and Shc, which play central roles in CCK's effects on acinar cell function. PMID- 16713447 TI - Sex-specific regulation of growth plate chondrocytes by estrogen is via multiple MAP kinase signaling pathways. AB - Both male and female rat growth plate cartilage cells possess estrogen receptors (ERs), but 17beta-estradiol (E(2)) activates protein kinase C (PKC) and PKC dependent biological responses to E(2) only in cells from female animals. PKC signaling can elicit genomic responses via mitogen activated protein kinase (MAPK) and E(2) has been shown to activate ERK MAPK in many cells, suggesting that MAPK may play a role in growth plate chondrocytes as well. We tested if E(2) increases MAPK activity and if so, whether the response is limited to female cells, if it is PKC-dependent, and if the mechanism involves traditional ER pathways. We also determined the contribution of MAPK to the biological response of growth plate chondrocytes and assessed the relative contributions of ERK, p38 and JNK MAPKs. Female rat costochondral cartilage cells were treated with E(2) and MAPK-specific activity determined in cell layer lysates. The mechanism of MAPK activation was determined by treating the cells with E(2) conjugated to bovine serum albumin (E(2)-BSA) to assess if membrane receptors were involved; stereospecificity was determined using 17alpha-estradiol; PKC and phospholipase C (PLC) dependence was determined using specific inhibitors; and the ER agonist diethylstilbestrol, the ER antagonist ICI 182780, and tamoxifen were used to assess the role of traditional ER pathways. E(2) regulation of ERK1/2 MAPK was assessed and the relative roles of ERK1/2, p38 and JNK MAPKs determined using specific inhibitors. E(2) caused a rapid dose-dependent activation of MAPK that was greatest in cells treated for 9 min with 10(-9) M hormone; activity remained elevated for 3 h. E(2)'s effect on MAPK was stereospecific and comparable to that of E(2)-BSA. It was insensitive to DES and ICI 182780, dependent on PKC and PLC, blocked by tamoxifen and it did not require gene transcription or translation. E(2) had no effect on ERK1 or ERK2 mRNA or protein but it caused a rapid phosphorylation of ERK1/2 at 9 min. Inhibition of ERK1/2 and p38 MAPK reduced the stimulatory effects of E(2) on alkaline phosphatase activity and [(35)S]-sulfate incorporation. These results suggest that E(2) regulates MAPK through a sex specific membrane-mediated mechanism that does not involve cytosolic ERs in a traditional sense and that ERK1/2 and p38 mediate the downstream biological effects of the hormone. PMID- 16713448 TI - Melanotransferrin stimulates t-PA-dependent activation of plasminogen in endothelial cells leading to cell detachment. AB - Tissue plasminogen activator (t-PA) is an extracellular serine protease that converts the proenzyme plasminogen into the broad-spectrum substrate serine protease, plasmin. Plasmin, one of the most potent pro-angiogenic factors, is a key element in fibrinolysis, cell migration, tissue remodeling and tumor invasion. In the present investigation, we assessed the impact of the truncated form of soluble melanotransferrin (sMTf) on plasminogen activation by t-PA and subsequent endothelial cell detachment. Co-treatment of human endothelial microvessel cells with plasminogen, t-PA and sMTf significantly increased plasmin formation and activity in the culture medium. Plasmin generated in the presence of sMTf also led to a 30% reduction in fibronectin detection within cell lysates and to a 9-fold increase within the corresponding cell medium. Moreover, the presence of sMTf increases EC detachment by 6-fold compared to cells treated only with plasminogen and t-PA. Although the addition of alpha(2)-antiplasmin completely prevented plasmin formation and EC detachment, epigallocatechin gallate, GM6001 and a specific antibody directed against MMP-2 prevented cellular detachment without interfering with plasminogen activation. Overall, these data suggest that the anti-angiogenic properties of sMTf may result from local overstimulation of plasminogen activation by t-PA, thus leading to subsequent degradation of the Fn matrix and EC detachment. PMID- 16713449 TI - Gene expression analysis of ELF-MF exposed human monocytes indicating the involvement of the alternative activation pathway. AB - This study focused on the cell activating capacity of extremely low frequency magnetic fields (ELF-MF) on human umbilical cord blood-derived monocytes. Our results confirm the previous findings of cell activating capacity of ELF-MF (1.0 mT) in human monocytes, which was detected as an increased ROS release. Furthermore, gene expression profiling (whole-genome cDNA array Human Unigene RZPD-2) was performed to achieve a comprehensive view of involved genes during the cell activation process after 45 min ELF-MF exposure. Our results indicate the alteration of 986 genes involved in metabolism, cellular physiological processes, signal transduction and immune response. Significant regulations could be analyzed for 5 genes (expression >2- or <0.5-fold): IL15RA (Interleukin 15 receptor, alpha chain), EPS15R (Epidermal growth factor receptor pathway substrate 15 - like 1), DNMT3A (Hypothetical protein MGC16121), DNMT3A (DNA (cytosine-5) methyltransferase 3 alpha), and one gene with no match to known genes, DKFZP586J1624. Real-time RT-PCR analysis of the kinetic of the expression of IL15RA, and IL10RA during 45 min ELF-MF exposure indicates the regulation of cell activation via the alternative pathway, whereas the delayed gene expression of FOS, IL2RA and the melatonin synthesizing enzyme HIOMT suggests the suppression of inflammatory processes. Accordingly, we suggest that ELF-MF activates human monocytes via the alternative pathway. PMID- 16713450 TI - Chemoprevention of nonmelanoma skin cancer. AB - Skin cancer is the most common cancer in human beings. The increased incidence of skin cancer has brought much attention to the process by which these tumors develop and how they can be prevented. Efforts have been made to educate the public about the importance of protecting skin from excessive ultraviolet light. Despite this work, the incidence of skin cancer continues to increase. Available compounds may be useful in the chemoprevention of skin cancer. Chemoprevention is defined as oral or topical use of dietary or pharmacologic agents to inhibit or reverse the development of cancer. Potential agents included are the retinoids; difluoromethylornithine; T4 endonuclease V; polyphenolic antioxidants, such as ( )-epigallocatechin gallate, found in green tea and grape seed extract; silymarin; isoflavone genestein; nonsteroidal anti-inflammatory drugs; curcumin; lycopene; vitamin E; beta-carotene; and selenium. Many of these agents are available over the counter as topical or oral preparations. LEARNING OBJECTIVE: At the conclusion of this activity, participants should be familiar with the chemopreventive agents and their efficacy, as well as any significant side effects associated with them. PMID- 16713451 TI - Toll-like receptors: applications to dermatologic disease. AB - Toll-like receptors are a recently identified group of receptors that are an important component of the immune system. Thus far, ten different receptors have been identified and have unique tissue distribution, ligand binding properties, cellular signaling pathways, and cytokine production profiles. Importantly, ligand binding has been shown to regulate both the adaptive and host immune response; thus, defects in this pathway have the potential to lead to increased susceptibility to infection and inflammatory dysregulation. In this article, the burgeoning literature pertinent to the discovery and signaling mechanisms are reviewed in addition to the discussion of the important role Toll-like receptors may play in the pathogenesis of numerous skin diseases. LEARNING OBJECTIVE: At the completion of this learning activity, participants should be familiar with the role of Toll-like receptors in host defenses and their relevance to dermatologic diseases. PMID- 16713452 TI - Development and reliability testing of a standardized questionnaire to assess psoriasis phenotype. AB - BACKGROUND: Psoriasis genetics researchers have utilized separate and widely differing survey instruments to capture clinical data to be utilized in genotype phenotype studies, which make comparison and collaboration studies among these researchers difficult. OBJECTIVE: The purpose of this study was to develop and validate a clinical survey instrument to facilitate future collaborative genotype phenotype studies among psoriasis genetics researchers. METHODS: The Delphi method was employed to obtain international consensus on components of the novel survey instrument. The survey was pretested for acceptability purposes, and then formally tested for reliability using 3 independent raters interviewing 48 subjects with psoriasis. RESULTS: Data showed high or moderately high agreement for questions relating to place of birth (85% to 100% in agreement), family origin (kappa = 0.48 -1.0), psoriasis history (kappa = 0.66-1.0), patient medical history (kappa = 0.76-1.0), distribution of lesions (kappa = 0.73-1.0), precipitating factors (kappa = 0.79-1.0), joint involvement (kappa = 0.74-.91), and treatment history, including use of oral retinoids, methotrexate, and etanercept (kappa = 0.73-1.0). Other parameters had lower degrees of agreement. LIMITATIONS: The time involved and the need for the rater to be a clinician with knowledge of psoriasis may preclude widespread use of this survey instrument. CONCLUSION: We developed a novel, reliable survey instrument that can be used to gather clinical information in a standardized manner from psoriasis patients participating in clinical and genetics research studies. PMID- 16713453 TI - Altered keratin 17 peptide ligands inhibit in vitro proliferation of keratinocytes and T cells isolated from patients with psoriasis. AB - BACKGROUND: Identification of critical autoantigenic T-cell epitopes is key to developing antigen-based therapies for autoimmune diseases, including psoriasis. Our previous work demonstrated that 3 peptides on keratin 17 are able to stimulate peripheral blood lymphocytes of HLA-DRB1*07-positive patients with psoriasis and to serve as immunodominant T-cell epitopes. OBJECTIVE: We sought to determine antagonistic altered peptide ligands to psoriatic T cells with a down modulatory effect in inhibiting keratinocyte proliferation. METHODS: Psoriatic altered peptide ligands were generated by single alanine residue substitutions at a critical T-cell receptor contact residue position. Antagonistic altered peptide ligands were identified by suppression screening of psoriatic T-cell activation and keratinocyte proliferation. RESULTS: Altered peptide ligands 119R and 355L can inhibit psoriatic T-cell activation more effectively than other altered peptide ligands, especially 355L, with inhibition of T-cell proliferation and the secretion of interferon gamma and interleukin 2 in parallel with the up regulation of interleukins 4 and 10 as well as transforming growth factor-beta. In coincubation assay, altered peptide ligands 119R and 355L can down-regulate the function of psoriatic T cells more effectively than wild-type epitopes solely, but less effectively than altered peptide ligands solely. In prepulse assay altered peptide ligand 119R can down-regulate the activation of psoriatic T cells more effectively than in coincubation but less effectively as compared with altered peptide ligand 119R only. Altered peptide ligand 355L was also shown to have a similar presentation. T-cell culture supernatants (1:100) from the concentrations (10 microg.mL(-1) and 100 microg.mL(-1) with 119R, 100 microg.mL( 1) with 355L) were more effective than the other ratios in inhibiting keratinocyte proliferation. LIMITATIONS: This study had a relatively small sample size (52 patients and 48 healthy controls). CONCLUSION: Our findings show that the altered peptide ligands 119R (VAALEEANTELEVKI) and 355L (ENRYCVQASQIQGLI) are capable of inhibiting proliferative responses of psoriatic T cells and keratinocyte proliferation in vitro, at least, with enhanced helper T cell type 2 polarization. Thus, to our knowledge, this article is the first report of the demonstration of therapeutic activity of altered peptide ligands derived from keratin 17. PMID- 16713454 TI - Resolution of endothelial activation and down-regulation of Tie2 receptor in psoriatic skin after infliximab therapy. AB - BACKGROUND: Psoriasis is a common dermatosis characterized by erythematous skin plaques and associated arthritis. Microvessels of the papillary dermis in psoriatic lesions are elongated, tortuous, and dilated, which contributes significantly to the proinflammatory response. Angiopoietin (Ang) 1 and 2 and their receptor, Tie2, are a family of growth factors recognized in inflammatory lesions to be critical for new blood vessel growth and maintenance, with recent studies suggesting tumor necrosis factor (TNF)-alpha-induced angiogenesis is in part mediated by the Tie2 receptor. The aim of this study was to evaluate the effect of anti-TNF-alpha therapy on angiogenic growth factor expression and on the cellular infiltrate in psoriatic lesional skin. METHODS: Sixteen patients with moderate to severe psoriasis and associated psoriatic arthritis (n = 13) received infliximab infusions (3-5 mg/kg) at baseline and at 2 and 6 weeks. Clinical assessments and skin biopsies were undertaken at baseline, and at 2 and 12 weeks. Ang 1, Ang 2, Tie2, and TNF-alpha messenger RNA expression were quantified by real-time polymerase chain reaction. Protein expression of vascular endothelial growth factor, Ang 2, Tie2, TNF-alpha, and the inflammatory infiltrate was determined using immunohistology. We conducted clinical assessments including Psoriasis Area and Severity Index, percentage body surface area, Arthritis Disease Activity Score, and Health Assessment Questionnaire. RESULTS: At baseline expression of Ang 1/2, vascular endothelial growth factor, Tie2, and TNF-alpha messenger RNA and protein were greater in preinvolved skin compared with uninvolved skin (P < .05). Infliximab produced a significant reduction in protein expression of Ang 2, vascular endothelial growth factor, and Tie2 (P < .001) along with a decrease in messenger RNA expression of Ang 1 (P < .045) and Tie2 (P < .021). This was paralleled by a significant reduction in the inflammatory infiltrate scores (P < .001) and platelet-endothelial cell adhesion molecule (CD31) expression (P = .001), suggesting deactivation of endothelial cell. There was a 93% mean reduction in Psoriasis Area and Severity Index (P = .001), and a significant reduction in Disease Activity Score 28 (P = .012) and mean Health Assessment Questionnaire scores by week 12. LIMITATIONS: This study involves a small number of patients. CONCLUSION: These results suggest infliximab is both effective and well tolerated in severe psoriasis, resulting in deactivation of endothelium and down-regulation of growth factor and cytokine expression, leading to a decrease in the cellular infiltrate and clinical improvement in psoriasis. Furthermore, the effect of infliximab on growth factor expression, in particular Tie2, supports previous in vitro work suggesting TNF alpha may be a major regulator of the Ang/Tie2 pathway. PMID- 16713455 TI - Methotrexate plus narrowband UVB phototherapy versus narrowband UVB phototherapy alone in the treatment of plaque-type psoriasis: a randomized, placebo-controlled study. AB - BACKGROUND: Combining phototherapy with topical and oral agents allows clinicians to treat recalcitrant psoriasis with reduced number of treatments and cumulative UV exposures. OBJECTIVE: This study was designed to determine the number of treatments necessary to clear plaque-type psoriasis when narrowband (NB) UVB is administered with methotrexate (MTX) or placebo in a randomized, controlled fashion. METHODS: MTX (15 mg/wk) or placebo was administered 3 weeks before standard NB UVB phototherapy was started. Treatments with the oral agent and phototherapy were continued until Psoriasis Area and Severity Index scores were reduced to less than 10% of the original scores or 24 weeks. Follow-up was performed until lesional scores returned to 50% of the original ones. RESULTS: A total of 24 patients were enrolled and 19 patients completed the study. Kaplan Meier analysis revealed that the median time to clear psoriasis in the MTX/NB UVB group was 4 weeks, which was significantly less than that for the placebo/NB UVB group. LIMITATIONS: Our sample size was relatively small (24 patients) with 5 dropouts. In addition, the study was conducted in skin types III to IV, Asian patients. Follow-up was limited to 4 to 6 months after completion of phototherapy. CONCLUSION: MTX pretreatment allows physicians to clear psoriasis in fewer phototherapy sessions than when phototherapy is administered alone. PMID- 16713456 TI - Clinical e-mail in an academic dermatology setting. AB - BACKGROUND: Use of e-mail among patients and physicians is limited by reservations over issues such as medicolegal aspects, reimbursement, and time management. OBJECTIVE: Our purpose was to identify the content of patient-related e-mails in an academic dermatology practice and determine whether clinical questions could be answered by e-mail. METHODS: The first 100 e-mails received each year that related to patients from January 1, 2000 to June 1, 2005 (plus any messages received in e-mail threads started in the original 100) were studied (N = 614). E-mails were sent by patients, potential patients, or physicians in reference to a patient. E-mails were divided into 8 categories on the basis of content. E-mails were subdivided as relating to new (patients who had never been seen in-person) or established patients. All clinical questions were categorized as to whether they were answered by e-mail. The average number of e-mails received per e-mail thread was tallied. RESULTS: E-mails were distributed as follows: clinical question from a physician (20%), clinical question from a patient (17%), appointment request (18%), request for referral to another physician (7%), prescription refill (3%), research inquiry (2%), thank-you correspondence (31%), other (17%). Percentages do not equal 100 because some e mails contained more than one subject. Clinical questions were more likely to be answered when posed by physicians (100%) than patients (70%; P = .001), and when from established (79%) versus new patients (60%; P = .02). There were fewer e mails per thread for queries from physicians (1.6 messages received) versus patients (2.2; P < .001) and for established (1.6) versus new patients (2.2; P < .001). LIMITATIONS: This study was limited to the experience of one dermatologist in an academic setting. CONCLUSION: E-mail broadens communication between patients and their dermatologist. E-mail may facilitate consultation with other physicians and management of patients with chronic disease. "Thank-you" responses engage a substantial amount of e-mail resources. PMID- 16713458 TI - Long-term follow-up of basal cell carcinomas treated with perilesional interferon alfa 2b as monotherapy. AB - BACKGROUND: Interferon alfa-2b (IFN) may be used to treat basal cell carcinoma (BCC) as an alternative to surgical or destructive methods. OBJECTIVE: The purpose of this study is to determine the long-term effectiveness of IFN treatment for BCC. METHODS: Fifty patients with 98 biopsy-proven primary superficial and nodular BCCs were treated perilesionally and intradermally with injections of IFN between 1985 and 1992. RESULTS: Clinical cures were noted in 95 of 98 BCCs (51 nodular and 44 superficial), with a mean follow-up period of 10.5 years (9 months to 18.5 years). Of these, 35 of the 50 patients, which would include 68 of the 98 tumors, were followed up for a minimum of 10 years, with an average follow-up of 13.5 years. The 3 lesions requiring further treatment were nodular type BCC. One of these lesions showed no response to treatment, whereas the other two responded, then recurred at 4 and 154 months. Of the 68 tumors followed up for a minimum of 10 years, cure rates were 96%. Overall data analysis using Kaplan-Meier estimates showed 98% success rates at years 5 and 10, and a 96% success rate at year 15. LIMITATIONS: Since 31% of patients could not be evaluated for more than 10-year follow-up, it is possible that the long-term cure rate is lower than that found in those available for evaluation. CONCLUSIONS: Treatment of superficial and nodular BCCs with perilesional IFN is an acceptable treatment that may provide benefits over other treatment modalities depending on patients' individual needs. In addition, on the basis of this study, results of IFN treatment for BCC are comparable to most other methods of tumor destruction. PMID- 16713457 TI - Imiquimod 5% cream monotherapy for cutaneous squamous cell carcinoma in situ (Bowen's disease): a randomized, double-blind, placebo-controlled trial. AB - BACKGROUND: We conducted a double-blind, placebo-controlled, randomized trial to evaluate the preliminary efficacy and safety of imiquimod 5% cream treatment for cutaneous squamous cell carcinoma (SCC) in situ. METHODS: In all, 31 patients with biopsy-proven cutaneous SCC in situ were randomly assigned to placebo (vehicle) (n = 16) or imiquimod 5% cream (n = 15) daily for 16 weeks. Patients were assessed at week 28 for the primary end point, resolution of cutaneous SCC in situ. RESULTS: Of the 31 patients enrolled, 3 dropped out. Intention-to-treat analysis revealed 11 of the 15 patients (73%) in the imiquimod group achieved resolution of cutaneous SCC in situ, with no relapse during the 9-month follow-up period; none in the placebo group achieved resolution (P < .001). Imiquimod 5% cream was generally well tolerated and there were no serious adverse events. LIMITATIONS: Topical imiquimod 5% cream has proven to be an effective treatment for cutaneous SCC in situ. However, studies to define the ideal dosing regimen and cost-effectiveness are required before it can be accepted as a recognized therapy. CONCLUSIONS: In this controlled trial, patients with cutaneous SCC in situ receiving topical imiquimod 5% cream as monotherapy experienced a high degree of clinical benefit compared with placebo. PMID- 16713459 TI - Treatment of basal cell carcinoma with curettage alone. AB - BACKGROUND: Although curettage and electrodesiccation (C&E) is widely used to treat basal cell carcinoma, whether electrodesiccation improves outcome is unknown. OBJECTIVE: We sought to compare cure rates of curettage alone with those of C&E. METHODS: We conducted a retrospective records review of patients treated with curettage alone at 5-year follow-up or longer that extracted data about tumor location, size, histologic subtype, biopsy specimen margin involvement, and recurrence, as well as data about the medical history of patients treated in a dermatology clinic in a tertiary-care academic medical institution. RESULTS: Biopsy-proven tumors (302) amenable to treatment with C&E and treated by a single investigator with curettage alone had a 5-year cure rate of 96.03%, with minimal complications (hypopigmentation, scarring). Tumors involving more than 50% of the deep edge of the shave biopsy specimen had an increased risk of recurrence. LIMITATIONS: This is a retrospective study based on historic controls. CONCLUSION: For nonaggressive basal cell carcinoma, curettage alone has a cure rate similar to the published rates for C&E. PMID- 16713460 TI - Infantile perineal protrusion. AB - Infantile perineal protrusion, a relatively newly recognized condition, is underreported in both the dermatologic and pediatric literature. The name "infantile perineal protrusion" has evolved based on the typical anatomic location, morphologic features, and prevalence in prepubertal children. It occurs in 3 settings: constitutional (sometimes genetic or familial); functional (after constipation, diarrhea, or other irritant exposure); or associated with lichen sclerosus et atrophicus. Recognition of infantile perineal protrusion by dermatologists and pediatricians has many implications regarding proper diagnosis and management. The condition may be mistaken for condyloma acuminata or as a sign of trauma, leading to an erroneous investigation of sexual abuse. In this article, we report two new cases and make a thorough review of the literature to elucidate the mechanisms, diagnosis, classification, and management to clarify this often misdiagnosed condition. PMID- 16713461 TI - Dermatophytes: diagnosis and treatment. PMID- 16713462 TI - Thrombocytopenia, multiple mucosal squamous cell carcinomas, and dyspigmentation. PMID- 16713463 TI - Use of artificial tanning products among young adults. AB - BACKGROUND: Neither the prevalence of sunless tanner use nor its impact on sunburning and tanning bed use has been evaluated in the United States. OBJECTIVE: We surveyed young adults in greater Boston to measure use of artificial tanning products, as well as recent history of sunburns and tanning bed use. METHODS: In July 2004, 448 individuals 18 to 30 years of age completed a brief questionnaire at universities, shopping venues, and parks. RESULTS: Twenty two percent of respondents used sunless tanning lotions in the preceding 12 months, and another 22% had not used them but would consider doing so in the coming year. Sunless tanning users were more likely to be female, younger, and more likely to report being severe burners. Both users and potential users were more likely to have sunburned during the summer and to have used tanning beds than those who neither used nor intended to use sunless tanning lotions, even after controlling for skin type. LIMITATIONS: The study was based on a non randomly selected sample in one city and was cross-sectional in nature. CONCLUSION: Our study raises the possibility that sunless tanning products do not decrease rates of sunburning or use of tanning beds. While safe alternatives to ultraviolet exposure are desirable, the potential risks of widely endorsing artificial tanning products must be considered. PMID- 16713465 TI - The frequency of reporting of race/ethnicity in case reports. PMID- 16713466 TI - As simple as black and white? PMID- 16713467 TI - How complicated are black and white? PMID- 16713468 TI - Skin color--in perspective. PMID- 16713469 TI - On the naming of people. PMID- 16713470 TI - Describing patients' "race" in clinical presentations should be abandoned. PMID- 16713472 TI - On the right track. PMID- 16713473 TI - Proposed policy on identification of race, ethnicity, or skin color in case reports and studies submitted to the Journal of the American Academy of Dermatology. PMID- 16713475 TI - Hand dermatitis. PMID- 16713476 TI - Medical pearl: Stereomicroscopy is a useful method to examine hair shaft anomalies. PMID- 16713477 TI - Pilot trial of 1% pimecrolimus cream in the treatment of seborrheic dermatitis in African American adults with associated hypopigmentation. AB - BACKGROUND: African Americans with seborrheic dermatitis may manifest associated hypopigmenation. Corticosteroids and antifungals are often used for treatment, yet chronic use of corticosteroids may be associated with skin atrophy, increased intraocular pressure, or further hypopigmenation. Pimecrolimus has been used successfully in a few patients with seborrheic dermatitis. OBJECTIVES: This open label, pilot trial assessed the efficacy and tolerability of pimecrolimus in the treatment of seborrheic dermatitis in African Americans with hypopigmentation. METHODS: Five African American adults with seborrheic dermatitis used a thin layer of pimecrolimus on the involved areas twice per day for 16 weeks. Clinical measures of improvement included erythema, scaling, and pruritus. Hypopigmentation was measured objectively using a mexameter. RESULTS: All participants noted a marked decrease in the severity of their condition. An improvement in hypopigmentation was also noted. For all indicators, the magnitude of improvement was most marked during the initial 2 weeks of treatment. LIMITATIONS: This was an open-label pilot trial limited to just 6 participants, only 5 of whom completed the study. CONCLUSIONS: Topical pimecrolimus cream may be an excellent alternative therapeutic modality for treating seborrheic dermatitis in African Americans, particularly in those with associated hypopigmentation. PMID- 16713478 TI - Diagnosis of lymphoma in the draining lymph node basin at the time of in-transit metastasis of cutaneous melanoma. AB - The association between cutaneous melanoma and non-Hodgkin's lymphoma (NHL) has been described. Herein we describe two cases in which lymphoma was detected in the draining lymph node basin at the time of the finding of in-transit metastasis of cutaneous melanoma. In both of these cases, sentinel lymph node biopsy at the time of the excision of the primary melanoma showed no evidence of metastatic melanoma or lymphoma. The temporal and spatial correlation of melanoma and NHL in these two cases suggests the possibility of a local failure in immune surveillance. PMID- 16713479 TI - Annually recurring erythema annulare centrifugum: a distinct entity? AB - Four patients presenting a peculiar clinical variant of erythema annulare centrifugum are reported. The lesions were clinically and histopathologically indistinguishable from classic superficial erythema annulare centrifugum but constant annual and seasonal recurrences for many years or decades were observed. No clear precipitating factor could be identified. No associated symptoms were present and the eruption regressed spontaneously after a variable period of days to months. Annually recurring erythema annulare centrifugum seems to represent a rare distinct clinical entity that has received little attention in literature. Clinicopathologic features of this peculiar clinical disorder and the differential diagnosis with other recurrent seasonal eruptions are reviewed. PMID- 16713480 TI - Unsuitability of organ donation from a patient with a history of melanoma? AB - A 52-year-old man with a history of melanoma presented to the emergency department with a massive intracranial hemorrhage. The patient deteriorated rapidly and was being considered as a potential organ donor. Three years before presentation, the patient had undergone wide excision of a 3.75-mm melanoma from his back with sentinel lymph node biopsy, which yielded negative findings. He had been well until the day of presentation. Although there are no specific guidelines for candidacy of organ donation from patients with a history of melanoma, there are several reports of donor-derived melanoma in organ transplant recipients, most with grave consequences. The literature relevant to this case is reviewed and discussed. PMID- 16713481 TI - Use of alefacept and etanercept in 3 patients whose psoriasis failed to respond to etanercept. AB - This article reports on the combined use of alefacept and etanercept in 3 patients whose psoriasis failed to respond adequately to etanercept. All patients maintained improvement for at least 8 weeks after completing 12 weeks of alefacept. No adverse events or infections were reported, and CD4+ T-cell counts remained above normal limits. PMID- 16713482 TI - Etanercept therapy in two patients with psoriasis and concomitant hepatitis C. PMID- 16713483 TI - Depigmentation following narrowband ultraviolet B phototherapy for psoriasis. PMID- 16713484 TI - Basal cell carcinoma responding to systemic 5-fluorouracil. PMID- 16713485 TI - Artifactual "pseudo-halo nevi" secondary to sunscreen application. PMID- 16713486 TI - Periungual myopericytoma. PMID- 16713487 TI - CXCR3 and CCR4 double positive tumor cells in granulomatous mycosis fungoides. PMID- 16713488 TI - Cutaneous involvement by Burkitt lymphoma. PMID- 16713489 TI - Self-limited variant of febrile ulceronecrotic Mucha-Habermann disease with polyclonal T-cell receptor rearrangement. PMID- 16713490 TI - Skin cancers in organ transplant recipients. PMID- 16713492 TI - Structure-function relationships in aquaporins. AB - The identification of members of the aquaporin family as the primary water channels of cell membranes has been followed up by an intense effort to determine how these channels work. Specifically, investigators have sought to learn why these channels are selective for water and how they exclude proton trafficking. Molecular-dynamics studies using elegant, extremely detailed computer models based on accurate crystallographic maps of the channels show the basis for the selectivity of the channel. Channel size, the location of hydrophobic amino-acid side chains, and specific interactions of water dipoles with a charged residue near the most constricted point of the channel indicate that water molecules travel in single file through the center of the channel, and that the orientation of water molecules is manipulated to prevent the formation of a water wire spanning the channel. Finally, the number of water molecules calculated to be aligned in single file in the channel constriction fits predictions based on classic studies of the osmotic permeability: diffusive permeability ratios in water-permeable membranes. PMID- 16713493 TI - Roles of aquaporins in kidney revealed by transgenic mice. AB - Transgenic mouse models of aquaporin (AQP) deletion and mutation have been instructive in elucidating the role of AQPs in renal physiology. Mice lacking AQP1 are unable to concentrate their urine because of low water permeability in the proximal tubule, thin descending limb of Henle, and outer medullary descending vasa recta, resulting in defective near-isosmolar fluid absorption in the proximal tubule and defective countercurrent multiplication. Mice lacking functional AQP2, AQP3, or AQP4 manifest various degrees of nephrogenic diabetes insipidus resulting from reduced collecting duct water permeability. Mice lacking AQP7 and AQP8 can concentrate their urine fully, although AQP7 null mice manifest an interesting defect in glycerol reabsorption. Two unexpected renal phenotypes of AQP null mice have been discovered recently, including defective proximal tubule cell migration in AQP1 deficiency, and cystic renal disease in AQP11 deficiency. AQPs thus are important in several aspects of the urinary concentrating mechanism and in functions unrelated to tubular fluid transport. The mouse phenotype data suggest the renal AQPs as targets for the development of aquaretics and potentially for therapy of cystic renal disease and acute renal injury. PMID- 16713494 TI - Familial neurohypophyseal diabetes insipidus--an update. AB - Although molecular research has contributed significantly to our knowledge of familial neurohypophyseal diabetes insipidus (FNDI) for more than a decade, the genetic background and the pathogenesis still is not understood fully. Here we provide a review of the genetic basis of FNDI, present recent progress in the understanding of the molecular mechanisms underlying its development, and survey diagnostic and treatment aspects. FNDI is, in 87 of 89 kindreds known, caused by mutations in the arginine vasopressin (AVP) gene, the pattern of which seems to be largely revealed as only few novel mutations have been identified in recent years. The mutation pattern, together with evidence from clinical, cellular, and animal studies, points toward a pathogenic cascade of events, initiated by protein misfolding, involving intracellular protein accumulation, and ending with degeneration of the AVP producing magnocellular neurons. Molecular research has also provided an important tool in the occasionally difficult differential diagnosis of DI and the opportunity to perform presymptomatic diagnosis. Although FNDI is treated readily with exogenous administration of deamino-D-arginine vasopressin (dDAVP), other treatment options such as gene therapy and enhancement of the endoplasmic reticulum protein quality control could become future treatment modalities. PMID- 16713495 TI - Hereditary polyuric disorders: new concepts and differential diagnosis. AB - The identification, characterization, and mutational analysis of genes coding for key proteins to the mechanisms of urine concentration provide the basis for understanding the 2 types of hereditary nephrogenic diabetes insipidus (NDI): a pure type characterized by loss of water only, and a complex type characterized by loss of water and ions. Patients with hereditary NDI bearing mutations in AVPR2, the gene coding for the arginine vasopressin 2 receptor, or in AQP2, the gene coding for the vasopressin-sensitive water channel, have a pure NDI phenotype with loss of water, but normal conservation of sodium, potassium, chloride, and calcium. Patients bearing inactivating mutations in 1 of the 5 genes (SLC12A1, KCNJ1, CLCNKB, CLCNKA, and CLCNKB in combination, or BSND) that encode the membrane proteins of the thick ascending limb of the loop of Henle have a complex polyuro-polydipsic syndrome with loss of water, sodium, chloride, calcium, magnesium, and potassium. The purpose of this article is to increase the general awareness of these congenital NDI patients to prevent severe episodes of dehydration and provide precise molecular diagnosis and treatment. PMID- 16713496 TI - Vaptans and the treatment of water-retaining disorders. AB - Hyponatremia is a frequent and symptomatic electrolyte disorder for which specific treatments have been lacking. Hyponatremia is attributable to nonosmotic vasopressin stimulation and continued increased fluid intake. In the past, peptidic derivatives of arginine vasopressin proved that blockade of vasopressin V-2 receptors served to improve hyponatremia, however, these antagonists had intrinsic agonistic activity, too. In the past decade, random screening of molecules uncovered nonpeptide, orally available vasopressin antagonists without agonistic properties. The agents show competitive binding to the vasopressin V-2 receptor at an affinity comparable with that of arginine vasopressin. Four antagonists have undergone extensive study. Three of these agents--lixivaptan or VPA 985; SR 121 463 B; tolvaptan or OPC 41,061--are specific V-2 antagonists whereas conivaptan or YM 087 is a V-1/V-2 mixed antagonist. In animal and clinical studies all of the agents were able to correct water retention and hyponatremia in a dose-dependent manner. There was no tachyphylaxis, even when the agents were given over many weeks. It is expected that the clinical use of the agents will lead to a major improvement in the treatment of hyponatremia. PMID- 16713497 TI - Acquired nephrogenic diabetes insipidus. AB - Nephrogenic diabetes insipidus (NDI) is defined as the inability of the kidney to concentrate urine owing to the insensitivity of the distal nephron to the antidiuretic hormone, arginine vasopressin. NDI can be either a congenital or an acquired disorder. Acquired NDI most commonly is secondary to drugs such as lithium or metabolic disturbances, such as hypokalemia and hypercalcemia. Disturbance of the aquaporin-2 shuttle is the underlying molecular basis of acquired NDI. NDI is diagnosed with the help of a water-deprivation test. Patients with the disorder will have a urinary osmolality of less than 300 mosm/kg H2O despite water deprivation. On administration of aqueous vasopressin, patients with NDI will show little or no increase in urine osmolality. Therapy consists of identifying and correcting the underlying disorder, or withdrawing the offending drug. Other treatment options that may be beneficial include diuretics, nonsteroidal anti-inflammatory drugs, decreased dietary solute intake, and desmopressin (DDAVP). PMID- 16713498 TI - Water intake and the neural correlates of the consciousness of thirst. AB - Thirst and resultant water drinking can arise in response to deficits in both the intracellular and extracellular fluid compartments. Inhibitory influences mediating the satiation of thirst also are necessary to prevent overhydration. The brain regions that underpin the generation or inhibition of thirst in these circumstances can be categorized as sensory, integrative, or cortical effector sites. The anterior cingulate cortex and insula are activated in thirsty human beings as shown by functional brain-imaging techniques. It is postulated that these sites may be cortical effector regions for thirst. A major sensory site for generating thirst is the lamina terminalis in the forebrain. Osmoreceptors within the organum vasculosum of the lamina terminalis and subfornical organ detect systemic hypertonicity. The subfornical organ mediates the dipsogenic actions of circulating angiotensin II and relaxin. Major integrative sites are the nucleus of the tractus solitarius, the lateral parabrachial nucleus, the midbrain raphe nuclei, the median preoptic nucleus, and the septum. Despite these advances, most of the neural pathways and neurochemical mechanisms subserving the genesis of thirst remain to be elucidated. PMID- 16713500 TI - MEG and TMS combined with EEG for mapping alcohol effects. AB - Magnetoencephalography (MEG) is a noninvasive method of studying magnetic fields from outside the skull that are generated by at least partially synchronized neuronal populations in the brain. The advantage of MEG over electroencephalography (EEG) is the transparency of the skull, scalp, and brain tissue to the magnetic fields, which facilitates easy localization of the cortical activity. In MEG, alcohol increased the relative power of the alpha rhythm and reduced the relative power of beta activity in parieto-occipital regions. In contrast, no changes were observed in EEG, indicating that these methods differently detect alcohol's action on the cortex. Furthermore, MEG and EEG also differently detected the effects of alcohol on cognition. Alcohol reduced magnetic and electric auditory N1 and mismatch negativity amplitudes. P3a amplitudes were also reduced in EEG but not in MEG, suggesting that different cortical areas are responsible for alcohol's action on involuntary attention. Transcranial magnetic stimulation (TMS) provides new possibilities for studying localized changes in the electrical properties of the human cortex, especially when combined with EEG. Different cortical areas can be stimulated and the subsequent brain activity can be measured, yielding information about cortical excitability and connectivity. Alcohol modulates EEG responses evoked by motor cortex TMS, the effects being largest at the right prefrontal cortex (assessed by minimum-norm estimation), meaning that alcohol changed the functional connectivity between motor and prefrontal cortices. Furthermore, alcohol decreases amplitudes of EEG responses after the left prefrontal stimulation of anterior parts of the cortex, which may be associated with the decrease of prefrontal cortical excitability. Taken together, MEG and TMS combined with EEG provide new insight into the focal actions of alcohol on the cortex with a temporal resolution of milliseconds, giving information different from that given by other brain imaging modalities. PMID- 16713501 TI - Does participation in an alcohol administration study increase risk for excessive drinking? AB - It has long been thought that research protocols involving alcohol administration may exacerbate problem drinking in alcoholic subjects following their participation in such a study. However, recent studies suggest that involvement in an alcohol administration study does not, in fact, have a negative impact on subsequent drinking behavior. In the present study, 27 non-treatment-seeking alcohol-dependent subjects and 32 social drinkers participated in an alcohol administration study designed to investigate the effects of repeated doses of alcohol on craving, mood, and alcohol-seeking behavior. The volume of alcohol administered to the subjects was calculated in such a way that their blood alcohol concentration would reach a peak of 0.08 g/dl midway through testing. Before their release, alcohol-dependent subjects were given feedback regarding their level of alcohol consumption and provided with information about the potential harmful effects of their drinking behavior. Percentage of days abstinent (PDA), drinks per drinking day (D/DD), and percentage of heavy drinking days (PHDD, defined as >or=4 drinks per occasion for females and >or=5 drinks per occasion for males) were recorded for the 6 weeks preceding laboratory testing and for the 6 weeks following participation in the study. The alcohol-dependent subjects exhibited a significant increase of 24% in PDA during the poststudy period compared to the prestudy period. They also decreased their D/DD by 2.4 drinks per occasion, and decreased their PHDD by 21.6%. There were no differences in PDA or D/DD for the social drinkers between pre- and poststudy periods. There was, however, a small but significant increase of 3.5% in PHDD for the social drinkers following laboratory testing. These data suggest that participation in an alcohol administration study does not put alcoholic subjects at risk for increased alcohol consumption following study participation. In fact, participation in such studies may actually precipitate at least a temporary decrease in alcohol consumption, especially when paired with a brief intervention session. Thus, non-treatment-seeking alcoholics can be safely included in alcohol administration studies to provide results that are most relevant to the population of interest. PMID- 16713502 TI - Alcohol levels in Chinese lactating mothers after consumption of alcoholic diet during postpartum "doing-the-month" ritual. AB - This study examined the effects of exposure to ethanol through cultural practices by lactating mothers. Specifically, the pharmacokinetics of alcohol in Chinese lactating mothers was investigated after they consumed chicken soup flavored with sesame oil and rice wine (CSSR), a typically prescribed diet during the postpartum "doing-the-month" period. Experimental findings were employed to estimate the potential ethanol dose to neonates and determine associated health risks. Twenty-three lactating mothers were examined. Informed consent was obtained from each subject. The target alcohol dosage was 0.3g/kg. Milk and blood samples were collected at fixed time intervals from each subject following exposure to CSSR, and alcohol levels were determined. Acute health risks to infants were estimated by comparing the potential infant dosage to an established criterion dose. Blood alcohol level peaked at 20 min after exposure to CSSR and decreased almost linearly thereafter. Alcohol in milk reached a plateau roughly at 20-40 min after exposure to CSSR and then decreased. Alcohol pharmacokinetics among subjects varied widely. The coefficients of variation in subject alcohol concentrations were 16.5-46.2% (mean, 30.0%) for blood and 32.8-57.6% (mean, 44.4%) for milk. Mean maximal alcohol concentration in blood (30.2+/-5.0 mg/dl) was achieved at 23.5+/-7.6 min and in milk (31.6+/-10.3 mg/dl) at 31.7+/-12.7 min. Potential infant doses were 3.0-58.8 mg (mean, 13.4 mg), and the predicted time required for milk alcohol level to return to zero level was 175 min. The acute health risks for infants exposed to alcohol through their mothers' milk under the current exposure scenario are low (hazard index<0.2). Nursing infants at least 3h after ingesting a diet containing alcohol would further reduce potential health risks. PMID- 16713503 TI - Apolipoprotein E4 genotype is not associated with short-term cognition deficits during alcohol withdrawal. AB - Aim of this prospective study was to investigate a possible association between the apolipoprotein E4 (ApoE4) genotype and clinically well-known cognition deficits during alcohol withdrawal. We examined 172 patients with alcohol dependence (137 men, 35 women) during withdrawal treatment. The ApoE genotype was determined in all patients using polymerase chain reaction. Cognitive function was assessed applying the c.I.-Test on day 0 (admission) and on day 7 of withdrawal treatment. Using Pearson's chi2 test we found no significant association between the ApoE4 genotype and cognition deficits for both dates (day 0: p=.463; day 7: p=.760). Moreover, multivariate logistic regression analyses revealed no significant association between presence of the ApoE4 allele and cognitive dysfunction. Even though ApoE4 plays an important role in alcoholism related brain atrophy and cognition deficits in demented as well as in nondemented healthy elderly people, this study provides no evidence for an association with short-term cognition deficits during alcohol withdrawal. PMID- 16713504 TI - Ethanol consumption increases rat stress hormones and adrenomedullary gene expression. AB - Numerous reports document altered drinking behavior following acute stressors but few describe physiological responses to acute stress of chronic ethanol consuming subjects. We tested rats' responses to 120-min foot restraint immobilization (Immo) after 1 week of liquid diet containing 5% wt/vol ethanol (ethanol-fed). Controls consumed isocaloric liquid diet ad libitum (adlib-fed) or in amounts equal to that of ethanol-fed subjects on the previous day (pair-fed). Each rat was implanted with a tail artery cannula on day 7 to allow remote blood collection before and during Immo on day 8. Plasma epinephrine (Epi); norepinephrine (NE); corticosterone (Cort); prolactin (PRL); adrenomedullary gene expression of catecholamine biosynthetic enzymes tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DBH), and phenylethanolamine-N-methyl transferase (PNMT); and TH protein levels were measured. Ethanol-fed rats had two to threefold higher basal plasma Epi and NE and tended to have increased Cort compared to adlib-fed or pair-fed rats. Immo increased Epi and NE in ethanol-fed rats more than twofold above those observed in controls, and also increased Cort more in ethanol-fed than in control rats. PRL was marginally affected. Ethanol potentiated the normal immobilization-induced increase in adrenomedullary TH, DBH, and PNMT messenger RNA (mRNA). TH protein increased only in ethanol-fed rats. Increased plasma catecholamine levels, adrenomedullary gene expression, and TH protein concentration in nonimmobilized ethanol-fed rats strongly suggest that ethanol consumption was itself a stressor, which potentiated the subsequent response to acute Immo. Moreover, the observed interaction of ethanol and stress on plasma catecholamine levels illustrates the importance of minimizing additional stressful stimuli when investigating ethanol's physiological effects. PMID- 16713505 TI - Ethanol familiarity and naltrexone treatment affect ethanol responses in rats. AB - In the present study, the effects of ethanol familiarity on the ability of naltrexone to alter ethanol palatability and consumption were examined. One group of rats was allowed continuous access to 10% vol/vol ethanol and water for 3 weeks. A second group received only water. At the end of this time, the groups were further subdivided and injected with either 3mg/kg naltrexone or saline (total of four groups; n=11-13 per group) before ethanol taste reactivity tests with 10% vol/vol ethanol and ethanol consumption tests. Results showed that naltrexone effectively decreased ingestive responding and increased aversive responding. Further, rats familiar with alcohol made more ingestive responses to 10% vol/vol ethanol. A significant interaction of drug treatment and familiarity was found in the data for aversive responses: naltrexone treatment produced more aversive responses in ethanol-familiar rats, whereas saline treatment resulted in fewer aversive responses in rats familiar with ethanol. Naltrexone treatment clearly reduced consumption of 10% vol/vol ethanol, although its effects were attenuated somewhat by ethanol familiarity. The present data indicate that both alcohol familiarity and naltrexone treatment affect ethanol reactivity and ethanol consumption in outbred rats. The interaction of naltrexone treatment and ethanol familiarity only for aversive reactivity and the lack of such an interaction for the consumption measures suggests that the mechanisms underlying ethanol reactivity and ethanol consumption may be dissociable at the neural level. PMID- 16713506 TI - Ethanol administration results in a prolonged decrease in blood ionized calcium levels in the rat. AB - Previous studies have shown that ethanol decreases the level of ionized calcium (iCa) in the blood, and appears to prevent a compensatory increase in parathyroid hormone level. We have shown, however, that the presence of ethanol interferes with the measurement of blood iCa by the most commonly used iCa analyzer. It is impossible to interpret ethanol-induced alterations in Ca-regulating hormone levels without accurate measurement of blood iCa, thus the purpose of this study was to determine if ethanol decreases blood iCa levels independent of methodological artifacts. The time course of ethanol's effect and the relationship between iCa and blood ethanol concentration (BEC) were also examined. Rats (n=22) received ethanol (1.5 or 3 g/kg body weight) or saline by intraperitoneal injection. Blood samples were obtained by tail nick at 0, 2, 6, 24, 48 h and 8 days postinjection, and analyzed for iCa, pH, and BEC. Blood iCa and pH were measured using the I-Stat Clinical Analyzer, whose performance is not affected by the presence of ethanol. Ethanol administration resulted in a decrease in blood iCa levels. The magnitude and time course of the decrease varied with dose of ethanol, being greater and more prolonged with the higher dose, and blood iCa levels were not fully recovered at 48 h postinjection. No significant relationship was found between individual iCa and BEC values. This study confirms that ethanol decreases blood iCa levels, independent of methodological artifacts. Prolonged disruptions in Ca homeostasis resulting from ethanol consumption could have implications for long-term bone health. PMID- 16713507 TI - Attention deficit hyperactivity disorder: nurses are important members of the team. PMID- 16713508 TI - Adolescent resilience: an evolutionary concept analysis. AB - Resilience is a concept that has enormous utility in nursing, although there is a need for concept clarity as it relates to adolescence. The purpose of this article is to describe the concept and apply the Evolutionary Model of Concept Analysis to resilience in adolescents. A literature search and an analysis of 22 articles specific to the adolescent population were completed. The literature review revealed adolescent resilience to be a composite of attributes that include the characteristics of the adolescent, sources of social support, and available resources. Implications for nursing and a proposed model of adolescent resilience are presented. PMID- 16713509 TI - Factors related to overweight and risk for overweight status among low-income Hispanic children. AB - The purpose of this research was to investigate the agent, host, and environmental factors that are related to overweight status in a sample of low income Hispanic toddlers who were enrolled in the Special Supplemental Nutrition Program for Women, Infants, and Children in a large southwestern city in the United States. Among children at risk for overweight status, the mothers' body mass index (BMI) and the number of servings of fat in the children's daily diet were positively associated with their weight/height percentiles whereas servings of protein in the daily diet and the children's reporting of hunger were negatively associated with their weight/height percentiles. Among overweight children, age was positively associated with their weight/height percentiles. No other variable in the ecological model was associated with normal weight or overweight children and their weight/height percentiles. Different approaches to dietary counseling and health promotion may be indicated based on a child's weight/height percentile or BMI. PMID- 16713510 TI - Attitudes toward physical activity in adolescents with cystic fibrosis: sex differences after training: a pilot study. AB - This study compared the attitudes of 16 adolescents (8 males and 8 females) with cystic fibrosis (CF) toward exercise and physical activity (PA) before and after a 6-week exercise program. Although the boys and girls had similar ages (12-18 years), the boys were fitter and leaner and had higher pulmonary function. Subjects reported both positive exercise attitudes of self and perceived attitudes of parents/friends that remained essentially unchanged after strenuous training. Boys reported higher vigorous activities at baseline than girls, but all subjects increased participation in very hard PAs after training. Girls had small but nonsignificant increases in PA at each (mild-vigorous) level. Results emphasize that adolescents with a mild to moderate lung disease can significantly increase PA in a nursing intervention. PMID- 16713511 TI - Children's and young adolescents' voices: perceptions of the costs and rewards of diabetes and its treatment. AB - Young peoples' perceptions of the costs and rewards related to diabetes may influence their understanding of, attitude toward, and adherence to positive diabetes-related behaviors. This qualitative descriptive research explores children's beliefs about the costs and rewards of Type 1 diabetes and its treatment via semistructured interviews. Children with diabetes attending a day camp comprised a purposive sample. These children were questioned using a researcher-developed interview guide. The results provide important insights into the perceptions of children and young adolescents with diabetes that may inform initiatives designed to foster positive diabetes-related behaviors. PMID- 16713512 TI - Self-care of young adolescents with type 1 diabetes. AB - The purpose of this study was to describe the universal and health deviation self care of adolescents with Type 1 diabetes and the associations of basic conditioning factors with universal and health deviation self-care. Subjects for this study were 152 adolescents aged between 11 and 15 years with a diagnosis of Type 1 diabetes. Data were collected in the home setting of each adolescent and his or her family. The mean universal self-care scores ranged from 66.62% to 90%. The overall mean for this sample was 75.37, indicating that these adolescents took care of their self-care needs 75% of the time. Health deviation self-care was a mean of 27.26, indicating more positive self-care behaviors and treatment adherence. Health deviation and universal self-care were significantly and positively related (r = .36, p < .001). Ethnicity and adolescent sex were statistically significant in predicting universal self-care. Adolescent age was statistically significant in predicting health deviation self-care. Health deviation self-care decreases with age, suggesting that early adolescence or late school age is an appropriate time for interventions to strengthen self-care behaviors. Furthermore, the interrelationship of the two types of self-care supports the potential for a synergistic effect of intervention. PMID- 16713513 TI - Let's keep children safe in cars. PMID- 16713514 TI - Daily physical activity and perception of condition severity among male and female adolescents with congenital heart malformation. AB - The purpose of this study was to examine among young patients with congenital heart malformation (CHM) the concordance between a cardiologist's definition of the severity of malformation and recommendations regarding physical activity and the patients' perceptions of their condition. Subjects included male (n = 55) and female (n = 45) adolescents aged between 12 and 18 years who have trivial (38%), mild (21%), or moderate (41%) CHM. Thirty-one percent of the patients rated their CHM as less severe whereas 15% rated their CHM as more severe than the cardiologist's definition. Although most patients engaged in physical activity appropriate for their condition, a noteworthy percentage engaged in activities more intense than those recommended by the patient's personal cardiologist. PMID- 16713516 TI - Can we improve cardiovascular risk prediction beyond risk equations in the physician's office? AB - BACKGROUND AND OBJECTIVES: Beyond a global estimation of the cardiovascular risk through the assessment of major risk factors and their integration in dedicated risk scales or equations, the use of specific markers provides additive prognostic information at an individual level, including predisposing factors, which are not included in the risk equations as well as the individual susceptibility to their long-term exposure. However, the majority of these markers require specific devices and skills, which are not widely available in primary care. METHODS: Some clinical and/or "low-cost" parameters are shown to be valuable risk markers, and their use could refine the risk estimation in a physician's office. Several epidemiologic studies suggest the heart rate, the pulse pressure and the ankle-brachial index are effective cardiovascular risk markers. The arms systolic pressure asymmetry could also be a useful marker of risk. RESULTS AND CONCLUSIONS: Through a general review, the authors evaluate the potential of these clinical markers, including their use in combination for more accurate risk determination. PMID- 16713517 TI - Results of diagnostic accuracy studies are not always validated. AB - BACKGROUND AND OBJECTIVE: Internal validation of a diagnostic test estimates the degree of random error, using the original data of a diagnostic accuracy study. External validation requires a new study in an independent but similar population. Here we describe whether diagnostic research is validated, which technique is used, and to what extent the validation study results differ from the original. STUDY DESIGN AND SETTING: All original diagnostic accuracy studies published in 1993 in a predefined set of journals were selected. Validation of these studies was assessed in the original article and in articles published within a period of 10 years, through a literature search and contacting the authors. RESULTS: None of the original studies reported any form of validation. Validation studies published later could be identified for 7 of the 11 original studies. Test characteristics were difficult to compare. Despite what was generally believed, not every validation study showed results inferior to the original. We found more studies that evaluated the test in a different population than in a similar one. CONCLUSION: Not every diagnostic accuracy study is validated. Diagnostic tests are more often repeated in different populations. PMID- 16713518 TI - A review finds that the autoreply e-mail function avoids problems of subject allocation concealment. AB - BACKGROUND AND OBJECTIVES: Using the autoreply e-mail function to avoid problems of subject allocation concealment. METHODS: A narrative review discussing technologic and methodologic issues in using autoreply e-mail to securely conceal subject allocation in randomized control trials. RESULTS: Autoreply e-mail can be successfully used to securely conceal subjects in a variety of randomized control trials. CONCLUSIONS: The allocation of subjects to intervention groups in randomized controlled trials should be achieved using methods that avoid both systematic bias and subject allocation transparency. The use of computer generated tables and the automatic e-mail reply system are suggested as a solution to ensuring subject allocation concealment. PMID- 16713519 TI - Period estimates of cancer patient survival are more up-to-date than complete estimates even at comparable levels of precision. AB - BACKGROUND AND OBJECTIVE: Period analysis provides more up-to-date estimates of cancer patient survival than traditional methods, but there is a trade-off between up-to-dateness and precision. Our objective was to compare the performance of period and complete analysis in terms of up-to-dateness and precision of survival estimates. STUDY DESIGN AND SETTING: Five-year relative survival data actually observed for patients diagnosed with 1 of 20 common forms of cancer in Finland in 36 overlapping 5-year periods between 1958-1962 and 1993 1997 were compared with period estimates and various variants of complete estimates of 5-year relative survival potentially available during these periods. RESULTS: At comparable levels of up-to-dateness, survival estimates from period analysis were more precise than survival estimates from complete analysis. At comparable levels of precision, period analysis provided more up-to-date survival estimates than did complete analysis. CONCLUSION: These results further encourage more widespread use of period analysis as a standard tool for up-to-date monitoring of cancer patient survival by population-based cancer registries. PMID- 16713520 TI - Multilevel modeling was a convenient alternative to common regression designs in longitudinal suicide research. AB - BACKGROUND AND OBJECTIVE: The prospective investigation of repetitive nonfatal suicidal behavior is associated with two methodological problems. Due to the commonly used definitions of nonfatal suicidal behavior, clinical samples usually consist of patients with a considerable between-person variability. Second, repeated nonfatal suicidal episodes of the same subjects are likely to be correlated. We examined three regression techniques to comparatively evaluate their efficiency in addressing the given methodological problems. STUDY DESIGN AND SETTING: Repeated episodes of nonfatal suicidal behavior were assessed in two independent patient samples during a 2-year follow-up period. The first regression design modeled repetitive nonfatal suicidal behavior as a summary measure. The second regression model treated repeated episodes of the same subject as independent events. The third regression model represented a hierarchical linear model. RESULTS: The estimated mean effects of the first model were likely to be nonrepresentative for a considerable part of the study subjects. The second regression design overemphasized the impact of the predictor variables. The hierarchical linear model most appropriately accounted for the heterogeneity of the samples and the correlated data structure. CONCLUSION: The nonhierarchical regression designs did not provide appropriate statistical models for the prospective investigation of repetitive nonfatal suicidal behavior. Multilevel modeling provides a convenient alternative. PMID- 16713521 TI - The DISABKIDS generic quality of life instrument showed cross-cultural validity. AB - OBJECTIVE: The aim was to test the performance of a cross-culturally developed disease-generic quality of life (QoL) measure for children with chronic diseases in a multinational clinical study. STUDY DESIGN AND SETTING: The study was conducted in pediatric hospitals across seven European countries in a sample of 1,153 children with seven chronic conditions, aged 8-16. The design included test and retest on child, parent, and clinical measures concerning health status, functioning, and quality of life. RESULTS: The condition-generic QoL measure showed satisfactory item and scale performance, as well as structural validity across the seven countries. There was a high agreement between the child and proxy assessment across the different countries in all subscales of the condition generic instrument, except for socioemotional dimensions in some countries. Gender, age, and severity-adjusted covariance analyses showed significant country differences in QoL. The discriminative potential of the condition-generic measure to distinguish between clinical characteristics was higher than that of a generic measures. CONCLUSION: The psychometric performance of both items and scales of the DISABKIDS condition oriented measure was good across cultures, despite a range of cross-cultural differences. The condition-generic measure performs better in discriminating with respect to clinical characteristics than the generic measures; however, results need replication in representative studies. PMID- 16713522 TI - A simple classification system was recommended for patients with restricted shoulder or neck range of motion. AB - OBJECTIVE: To construct an empirical classification of patients with shoulder complaints, and then to investigate the relationship between the empirical classification and the setting in which the patients were recruited, their demographic and clinical characteristics, and the original diagnostic categories. STUDY DESIGN AND SETTING: A latent class analysis was performed on the combined data of two previous studies. RESULTS: Four clusters of patients emerged, one with patients who have a small chance of any restriction of the motion of the shoulder and a moderate chance of restriction of the motions of the neck, a second with patients who have a high chance of restriction of the motions of the shoulder and a moderate chance of restriction of motions of the neck, a third with patients who have a low to moderate chance of restriction of all motions, and a fourth with patients who have a high chance of restriction of all of the motions. Patients recruited from a clinic for rheumatology and rehabilitation and from the orthopedic clinic were more present in the second cluster. Patients in the third cluster were on average younger than those in the other clusters. There appeared to be little agreement between the clusters found and the classification of patients according to the original diagnostic categories. CONCLUSION: Patients experiencing shoulder pain can be classified in a simple way into four categories, reflecting the distinction between problems of the shoulder and those of the neck. A simple classification rule is proposed with which almost all patients can be classified into the four clusters. PMID- 16713523 TI - A brief measure of perceived understanding of informed consent in a clinical trial was validated. AB - BACKGROUND AND OBJECTIVE: To develop and evaluate an Informed Consent Questionnaire (ICQ) for measuring self-reported perceived understanding of informed consent in a randomized clinical trial. METHODS: The study was embedded in a Department of Veterans Affairs randomized clinical trial of Gulf War veterans' illnesses (CSP#470). The trial was initiated in May 1999 at 20 hospitals and concluded in September 2001; 1,092 participants were enrolled and followed for 12 months. The reliability and validity sample included 1,086 participants evaluated at baseline, 906 at 3 months, 929 at 6 months, and 910 at 12 months. The psychometric evaluations included tests of acceptability (based on missing data, endorsement frequencies, and floor/ceiling effects), item reduction, internal consistency, and construct validity (based on Cronbach's alpha coefficients, item-total correlations, and principal components analysis). RESULTS: The ICQ had >5% missing information on some questions at baseline, indicating poor acceptability prior to the initiation of the trial; however, the scale had good acceptability at each of the follow-up visits. Psychometric evaluation following standard item reduction techniques confirmed the reliability and validity of a four-item subscale of the ICQ (ICQ-4). CONCLUSIONS: The ICQ-4 is a simple and psychometrically sound self-report measure of perceived understanding of informed consent. PMID- 16713524 TI - Interactive triage simulator revealed important variability in both process and outcome of emergency triage. AB - BACKGROUND AND OBJECTIVES: (1) to evaluate the performance of emergency department triage; (2) to explore the variability of the triage process; and (3) to examine the reliability of a four-level triage scale, using an interactive triage simulator. METHODS: We developed 22 interactive computerized vignettes describing patients presenting at the Emergency Department. Each vignette displayed the presenting complaint and offered the possibility to ask questions and obtain vital signs before deciding on the triage severity rating. The vignettes were rated twice by 45 nurses and 8 physicians. RESULTS: (1) The concordance between the observed triage decision and an expert-attributed emergency level was perfect in 58% of the situations. Triage acuity was overestimated in 11%, and underestimated in 31%. (2) There was a wide variability in the triage process across observers and vignettes. The mean number of questions varied from 1.77 to 18.95 across individuals, and from 3.96 to 11.60 across vignettes. (3) Finally, the test-retest reliability of our instrument was good (weighted kappa = 0.82) but the interrater reliability was moderate (weighted kappa = 0.41). CONCLUSIONS: The computerized triage simulator is an innovative tool to evaluate the process and the performance of triage and to evaluate the reliability of a triage instrument. PMID- 16713525 TI - Self-reported smoking cessation interventions were not associated with quitting in older women. AB - OBJECTIVE: To determine the extent to which smoking cessation interventions are used in the community and their relative success in older women. STUDY DESIGN AND SETTING: Prospective cohort study located in 23 general practices in England, Scotland. and Wales and including 3,622 women aged 60 to 79 years at recruitment from the British Women's Heart and Health Study. RESULTS: Of the 370 smokers at baseline, 77 (21%) had stopped smoking at 3 years, reducing the prevalence of smoking from 10.2% to 8.8%. Women who were light smokers (<10 per day) at baseline were more likely to have quit than heavier smokers. The most common interventions reported by women who smoked at baseline were advice from a general practitioner (GP), family or friend's advice, and use of nicotine replacement, but 42% of the women reported having no intervention to help them to stop smoking. None of the individual interventions were associated with increased odds of quitting smoking over the follow-up period. Women reported finding a GP's advice, referral to a clinic, and nicotine replacement more useful than complementary treatments (acupuncture and hypnotherapy) or advice from family and friends. CONCLUSION: Smoking cessation rates in this study were in line with national targets and surveys. Self-reports of ever receiving interventions were not associated with quitting smoking among these older women. Different and innovative approaches to smoking cessation may need to be developed, and evaluated in future randomized clinical trials, for individuals who have smoked for a long period of time and for whom other smoking cessation interventions have not proved successful. PMID- 16713526 TI - Asking older people about fear of falling did not have a negative effect. AB - BACKGROUND AND OBJECTIVE: To assess whether completing a questionnaire on risk of falling could affect outcome measures: fear of falling, reported falls, and health service contacts in older people (panel conditioning). METHODS: We used a postal questionnaire to assess the effect on falls risk of implementing falls injury prevention guidelines within a single locality in outer London, UK. We compared responses for the baseline and 6-month follow-up surveys with those for a fresh survey. The latter was sent to a new pool of subjects drawn from the same population, and was sent only once; timing coincided with the follow-up survey. RESULTS: At baseline, we received 498 responses for 1,000 (50%) surveys sent; of these, 358 (72%) subsequently returned the follow-up survey. For the fresh survey, we received 1,261 out of 2,000 (61%) responses to the fresh survey. The odds ratio for the effect of panel conditioning on fear of falling was 0.92 (95% confidence interval CI = 0.64-1.33), within our predefined limit for equivalence. Odds ratios for the effect on reported falls and health service contacts were 0.87 (95% CI = 0.59-1.29) and 0.75 (95% CI = 0.55-1.02), respectively. CONCLUSION: The proportions of subjects who feared falling in the follow-up survey and in the fresh survey were equivalent. Reduced reporting of falls and health service use in the follow-up survey suggest that the potential for panel effects cannot be ignored. PMID- 16713527 TI - Print format and sender recognition were related to survey completion rate. AB - OBJECTIVE: To assess whether print format (single-sided vs. double) and sender recognition (known vs. unknown) affect response and completion rates among physician survey respondents. STUDY DESIGN AND SETTING: Postal survey of 399 members of the Canadian Association of Emergency Physicians; 2 x 2 factorial design. RESULTS: Response rate was 69.4%. Single-sided printing yielded 7.4% (odds ratio OR = 1.41; 95% confidence interval CI = 0.90-2.20; P = 0.13), and a known sender yielded a 6.3% greater response rate (OR = 0.73; 95% CI = 0.47-1.14; P = 0.16). Overall item completion was 98.2%; items missed per respondent ranged from 1 to 14 out of 50. Print format and sender recognition interacted in predicting completion rate (OR = 13.33; 95% CI = 3.10-57.4; P = 0.001); completion was higher for double-sided printing with an unknown sender, and for single-sided printing with a known sender. Completion was also lower when response came after later mailouts (chi2(2) = 10.13; P = 0.006). CONCLUSIONS: Print format and sender recognition both yielded 6%-7% (nonsignificant) response rate differences. Survey completion rate varied even when overall item completion was high. Completion rate was useful for identifying subgroups likely to provide incomplete data (i.e., late responders), and may provide important information for subsequent surveys. Combining factors that on their own improve survey response may have unexpected consequences. PMID- 16713529 TI - Adherence to antihypertensive medication assessed by self-report was associated with electronic monitoring compliance. AB - BACKGROUND AND OBJECTIVE: Clinicians often find it difficult to differentiate between subobtimal adherence and nonresponse to medical treatment if a patient's clinical condition does not improve. This study aimed to evaluate an adherence self-report tool to assess adherence to blood pressure lowering medication. STUDY DESIGN AND SETTING: Participants were 245 patients with uncontrolled hypertension from 21 general practices taking part in a randomized trial. Prior to randomization, participants were asked to indicate which of six descriptions (adherence levels) would most closely describe their medication taking and were given electronic medication monitors for 30 days. We used multivariable regression analysis to compare the adherence self-report tool with electronic monitoring. RESULTS: There was strong evidence that a drop in one adherence level from level 1 (highest level) to level 2 and from level 2 to level 3-6 (six being the lowest) combined was associated with a decrease in timing compliance of approximately 5% (test for trend: P = 0.0004). CONCLUSION: Adherence assessed by a brief self-report tool was associated with timing compliance obtained through electronic monitoring in patients taking blood pressure lowering medication. Further research is required to investigate the validity of this tool in patients with lower adherence levels and in a nontrial population. PMID- 16713530 TI - Mechanisms and modeling of waste/cement interactions International Workshop, May 8 to 12, 2005, Meiringen, Switzerland. PMID- 16713528 TI - Psychometric properties of the revised Piper Fatigue Scale in Dutch cancer patients were satisfactory. AB - OBJECTIVE: To examine the psychometric properties of the revised Piper Fatigue Scale (PFS) in Dutch cancer patients. STUDY DESIGN AND SETTING: Participants were 64 patients with lung (any stage, nonsurgery) and breast (any stage) cancer, selected for curative high-dose radiotherapy (> or =50 Gy). Prior to radiotherapy, patients completed the revised PFS (translated into Dutch), the Multidimensional Fatigue Inventory (MFI), and the Rotterdam Symptom Checklist (RSCL). Reliability and construct and criterion validity of the PFS were investigated. RESULTS: Overall, the structure of the Dutch revised PFS appeared appropriate, with corrected item-subscale correlations being higher than the correlations of the same item with the three other subscales; furthermore, internal consistency was excellent (Cronbach's alpha > or =0.90). Lung cancer patients were significantly more tired then breast cancer patients, supporting construct validity. Criterion validity was also satisfactory, with highest correlations of PFS total fatigue with the MFI subscale general fatigue (0.84) and RSCL overall quality of life score (0.74). Internal consistency was similar in Dutch and U.S. breast cancer patients; fatigue scores were lower on several PFS subscales in the Dutch study population. CONCLUSION: Psychometric properties of the Dutch version of the revised PFS, as tested in cancer patients prior to radiotherapy, were satisfactory. PMID- 16713531 TI - SDF-1alpha-mediated tissue repair by stem cells: a promising tool in cardiovascular medicine? AB - Circulating smooth muscle progenitor cells have been identified as a source for neointimal smooth muscle cells after various types of injuries to the vessel wall contributing to neointimal hyperplasia, implying a fundamental role of these progenitor cells in the vascular response to injury. Recent studies have provided insight into the molecular mechanisms of mobilization and local recruitment of smooth muscle progenitor cells. The CXC chemokine SDF-1alpha and its receptor CXCR4 have been identified as the central signaling axis regulating the homing of smooth muscle progenitor cells into the injured vessel wall. This review discusses the unique biologic functions of this chemokine and its contribution to stem-cell-based vascular repair and disease. PMID- 16713532 TI - Nitric-oxide-induced vasodilatation: regulation by physiologic s-glutathiolation and pathologic oxidation of the sarcoplasmic endoplasmic reticulum calcium ATPase. AB - Nitric-oxide (NO)-induced vasodilatation is impaired in patients with a variety of cardiovascular diseases. Cyclic GMP, the principal mediator of NO-induced smooth muscle relaxation, usually functions normally, and the impairment in endothelial-mediated vasodilatation is attributed to increased oxidants that diminish NO bioactivity. In investigating the mechanisms involved, we found that independently of cyclic GMP, NO stimulates the uptake of cytosolic Ca(2+) via the sarcoplasmic reticulum Ca(2+) ATPase (SERCA), thereby relaxing vascular smooth muscle by lowering intracellular free Ca(2+). Nitric oxide was found to do so by reacting with superoxide to form peroxynitrite, which in turn caused glutathione (GSH) to bind to SERCA cysteine thiols. Most of the GSH was bound to the most reactive thiol on SERCA cysteine-674, and mutation of this residue prevented stimulation of Ca(2+) uptake by NO. In atherosclerotic arteries, we found that NO no longer stimulated SERCA activity because of the irreversible oxidation of the cysteine-674 thiol. These studies not only demonstrate a novel mechanism of NO induced vasodilatation, but also provide an explanation as to how chronically increased levels of oxidants associated with disease impair vasodilatation in diseased arteries. PMID- 16713533 TI - Survivin regulation of vascular injury. AB - Survivin is a protein member of the inhibitor of apoptosis family that has attracted attention for its dual essential roles in the regulation of cell division and the control of apoptosis. Although extensively characterized as a tumor gene and exploited for new opportunities in cancer diagnostics and therapeutics, fresh experimental evidence has uncovered a critical role of survivin in vascular cell responses in vivo. Dynamically embedded in broad signaling pathways controlling smooth muscle cell adaptation, survivin not only controls vascular wall homeostasis but also acts as a critical upstream regulator of gene expression and second mediators. PMID- 16713534 TI - Endothelial dysfunction and impaired L-arginine transport in hypertension and genetically predisposed normotensive subjects. AB - The term endothelial dysfunction (ED) generally refers to a pathological circumstance in which normal endothelial regulation of vasomotor tone is disturbed. ED has been described in a broad range of cardiovascular and related disease states. In association, deranged endothelial function also has the potential to result in abnormal platelet aggregation and monocyte adhesion. Over the past decade, it has become apparent that reduced availability of nitric oxide is a key mechanism for ED. A range of biochemical disturbances, including depressed L-arginine, have been shown to be associated with ED, and recent evidence indicates this may be genetically determined. PMID- 16713535 TI - Dependence of proliferating dedifferentiated vascular smooth muscle contraction on Rho-Rho kinase system. AB - Vascular smooth muscle cells (VSMCs) are not terminally differentiated and, owing to their remarkable plasticity, can change to a dedifferentiated state in response to vascular injury. Our understanding of the contractility of VSMCs is mainly based on the data obtained from normal adult animals. However, to obtain a better understanding of the abnormal contractility seen in the vascular diseases such as hypertension and vasospasm superimposed on atherosclerosis, it is important to also know the contractility of proliferating dedifferentiated VSMCs. To this end, we studied the contractility of cultured VSMCs that undergo dedifferentiation similar to that induced by vascular injury. There are only a few reports in which the contractility of cultured VSMCs has been extensively studied. We established a method to investigate the contractility of the cultured VSMCs and determined that their contraction is dramatically changed to be more dependent on the Rho-Rho kinase system but less dependent on the PKC-CPI-17 (protein kinase C-potentiated protein phosphatase 1 inhibitory protein)-mediated pathway. In this review, we focus on the contractility of the cultured VSMCs as a model of the proliferating dedifferentiated VSMCs. PMID- 16713536 TI - Are intra-aortic hemopoietic cells derived from endothelial cells during ontogeny? AB - The aorta is recognized as an intraembryonic site that produces adult-type hemopoietic stem cells. A corpus of data indicates that hemopoietic cells arranged as clusters attached to the aortic floor derive from an endothelial intermediate. This review reports on experimental approaches carried out in the avian embryo to establish the developmental history of the aortic endothelium and trace the origin of associated hemopoietic cells. PMID- 16713537 TI - An orthotopic colon cancer model for studying the B7-H3 antitumor effect in vivo. AB - We established an orthotopic animal model of colon cancer in mice and applied this model to study the antitumor effects of B7-H3, the newest member of the B7 family of costimulatory molecules. Colon-26 murine colon adenocarcinoma cells were inoculated into the cecal subserosum of mice to induce colon tumor growth. The tumor growth rate and the survival time of the mice were observed. A stable B7-H3 transfected Colon-26 cell line was established and the immunogenic effect was investigated. All mice implanted with wild-type tumor cells had tumor growth in the colon and died. The mean survival rate was 23 days. Mice implanted with C26-B7-H3 had a significantly prolonged survival time of 38 days. Our data suggest that B7-H3 exerts an antitumor effect on adenocarcinoma of the colon and may be considered as an adjuvant immunotherapy in the treatment of colon cancers. Our orthotopic animal model of colon cancer in mice could be applied to in vivo experimental studies of colon cancer. PMID- 16713538 TI - Analysis of postsurgical complications in 75 living liver transplantation donors. AB - Seventy-five living donor liver hepatectomies were performed at our transplantation center between April 1990 and December 2004. We collected the data from patient charts, files, and the Baskent University Liver Registry. There were 39 male and 36 female donors (mean age, 35.1 +/- 9.3 years). We have performed 29 (38.6%) left hepatic lobectomies, 18 (24%) left lateral segmentectomies, 26 (34.6%) right lobectomies, and two (2.6%) donors had simultaneous living donor nephrectomy plus left lobe hepatectomy. The mean remnant liver volume was 598 +/- 168 cm(3) (range, 410-915 cm(3)). The mean percentage of remnant liver for the donor was 55.2%. Mean postoperative hospital stay was 10 +/- 4.4 days. After surgery, there was no mortality or reoperation. We saw 15 (20%) postsurgical complications in 14 donors. Intra-abdominal collection was seen in five (6.6%) patients. Biliary leak was seen in four patients. Portal vein thrombosis was seen in one patient, and a pulmonary embolus developed in one liver donor. Patient safety must be the primary focus in living donor liver transplantation. These donors face significant risks, including substantial morbidity and death. More experience, improved surgical techniques, and meticulous donor evaluation will help minimize morbidity and mortality for both living liver donors and recipients. PMID- 16713539 TI - An approach to analyze mechanisms of intestinal adaptation following total proctocolectomy. AB - We hypothesized that epithelial cells of the remnant small intestine display "colonic" phenotype after total proctocolectomy. The aims of the present study were to identify preferentially expressed molecules in the colon or in the small intestine and to evaluate mRNA levels of those in the ileal pouch. Differential gene expression was investigated between the small intestine and the colon by using cDNA microarray and was confirmed by Northern blotting. Expression of three colonic mRNAs (3-hydroxy-3-methylglutaryl-coenzyme A synthase 2, deleted malignant brain tumors 1, carcinoembryonic antigen-related cell adhesion molecule 1) and one "small intestinal" (microsomal triglyceride transfer protein) mRNA were compared between the control and the ileal pouch mucosae by quantitative reverse transcriptase-polymerase chain reaction. Seventy-four clones were differentially expressed with more than a threefold difference. Differential expression was confirmed in all mRNAs examined, including 3-hydroxy-3 methylglutaryl-coenzyme A synthase 2 and microsomal triglyceride transfer protein. The mucosal expression of carcinoembryonic antigen-related cell adhesion molecule 1 mRNA in the ileal pouch was enhanced in humans. The remnant ileum develops some, but not all, colonic phenotype after total proctocolectomy. Comparative study of epithelial gene expression between the small intestine and the colon enables us to analyze mechanisms of intestinal adaptation after total proctocolectomy. PMID- 16713540 TI - Pancreatic cancer in sweden 1980-2000: a population-based study of hospitalized patients concerning time trends in curative surgery and other interventional therapies. AB - Progress has been made during the last few decades in the treatment of patients with pancreatic cancer. In this population-based study, the time trends in curative surgery and the choice of palliative invasive therapies in Sweden over two decades are analyzed. Patients treated for pancreatic carcinoma in Sweden during 1980-2000 were identified in the Swedish Hospital Discharge Register and the Cancer Register. These data were matched with those in the Register of Causes of Death in Sweden. Data were identified and analyzed for 16,758 patients for three periods: 1980-1986 (n = 5775), 1987-1993 (n = 6096), and 1994-2000 (n = 4887). The rate of pancreatic resection increased 7.2%, 10.9%, and 15.1% (P < 0.0001) during the three respective periods. Palliative interventions decreased from 46.8% in the first period to 41.7% in the last period. On comparing the first and the last periods, biliary bypass operations were found to decrease (from 45.9% to 18.1%), as well as gastric bypass procedures (from 33.8% to 22.8%; P < 0.0001). Interventions by percutaneous transhepatic cholangiography (PTC) remained constant (10%-11%). Endoscopic therapy increased from 10.8% to 49.0%, as did the number of procedures per patient, from 1.3 to 1.7 (P < 0.0001) in the first and last periods, respectively. In 1980, the mean hospital stay was 40 days after resection and 30 days after palliative intervention. In 2000, the corresponding numbers were 26 days and 18 days (P < 0.001), respectively. During the past two decades, the rate of pancreatic resections in Sweden increased significantly. There was also a dramatic drop in palliative open surgery and a simultaneous increase in endoscopic interventions. Hospital stays decreased by more than a third. PMID- 16713542 TI - Single center prospective randomized trial of laparoscopic Nissen versus anterior 90 degrees fundoplication. AB - Although Nissen fundoplication is a very effective treatment for gastroesophageal reflux, it is associated with a small incidence of troublesome postoperative side effects. To prevent this, progressive modification of surgical techniques has led to the development of an anterior 90 degrees fundoplication. We undertook a prospective randomized trial to compare this procedure with Nissen fundoplication to determine whether it would achieve a better clinical outcome. Patients presenting to a single center for primary laparoscopic antireflux surgery were randomized to undergo either an anterior 90 degrees fundoplication (n = 40) or a Nissen fundoplication without division of the short gastric vessels (n = 39). Clinical questionnaires were used to assess outcome at 1 month, 3-6 months, and 12 months. Both patients and the clinical interviewer were masked as to which procedure was performed. Follow-up with endoscopy, esophageal manometry, and pH monitoring was also undertaken. Operating time was similar for the two procedures (60 minutes for anterior vs. 55 minutes for Nissen fundoplication). Early postoperative complications were more common after Nissen fundoplication (18% vs. 5%). Two patients underwent laparoscopic reoperation for recurrent reflux after anterior 90 degrees fundoplication, and four underwent laparoscopic reoperation after Nissen fundoplication (dysphagia, 3 patients; acute hiatus hernia, 1 patient). One year after surgery, dysphagia and other wind-related side effects were less common after anterior 90 degrees fundoplication. Control of reflux symptoms and satisfaction with the overall outcome was similar for the two procedures. Anterior 90 degrees fundoplication is followed by fewer side effects than Nissen fundoplication. This advantage is offset by a greater likelihood of reflux recurrence. However, this does not diminish patient satisfaction. PMID- 16713541 TI - Is adjuvant 5-FU-based chemoradiotherapy for resectable pancreatic adenocarcinoma beneficial? A meta-analysis of an unanswered question. AB - The objective of this study was to determine the effect, if any, on survival of adjuvant 5-FU-based chemoradiotherapy following pancreaticoduodenectomy for pancreatic carcinoma. A systematic review of the published literature was undertaken. Survival estimates were derived from published reports. Five prospective studies (4 level I, 1 level II) with a total of 607 (229 surgery only; 378 surgery-adjuvant) patients followed for survival met selection criteria. Two-year survival ranged from 15%-37% in the surgery only group and 37% 43% in the surgery and adjuvant groups. The survival advantage (absolute difference) ranged from 3%-27% and no individual study achieved statistical significance (5%). Although clinical heterogeneity existed in surgery-alone control groups with regard to trial date, no statistical heterogeneity was detected (P = 0.459, chi2 test), allowing pooling of survival data. Using a fixed effects model, the summary estimate showed an absolute 2-year survival benefit with adjuvant therapy of 12% (95% CI, 3%-21%, P = 0.011). Trials after 1997 (n = 3) indicated a survival benefit of 8% to patients receiving adjuvant therapy (95% CI, -3-18%, P = 0.145). The result was not statistically significant, and there was no evidence of heterogeneity (P = 0.626, chi2 test). Summary estimates were unchanged when the analysis was performed with a random effects model. 5-FU based chemotherapy with radiotherapy given after resection imparts a small overall survival benefit of 2 years. The benefit of 5-FU-based adjuvant therapy, however, has declined in recent years, and its significance remains unproven in the context of current diagnostic and surgical practice. PMID- 16713543 TI - Role of glucocorticoid receptor in serosa-involved gastric carcinoma after gastrectomy. AB - Glucocorticoid receptor (GR) was first found in the cytosol of gastric cancer tissue more than 15 years ago. At present, most gastric cancers are diagnosed at the advanced stage. To elucidate the role of GR in gastric cancer, the GR levels of the cancer tissue of 75 consecutive patients with grossly serosa-involved gastric carcinoma were determined by the dextran-coated charcoal method. The clinicopathologic characteristics and long-term survival duration were compared in patients with GR-positive and GR-negative cancer cells. We found that GR could be detected in the cytosol of cancer cells in 31 (41.3%) of the gastric cancer patients with a median concentration of 18.5 (range, 1.03-73.9) fmol/mg protein. No significant differences could be found in any clinicopathologic characteristic between the patients with GR-positive and GR-negative cancers. After multivariate analysis, gross Borrmann's type, metastatic lymph node number, and GR positivity were the independent prognostic factors after gastrectomy for serosa-involved gastric carcinoma. GR-positive gastric cancer had a worse survival rate than GR negative gastric cancer. Multimodality adjuvant therapies should be considered in patients with GR-positive serosa-involved gastric carcinoma. PMID- 16713544 TI - Identification of Helicobacter pylori biofilms in human gastric mucosa. AB - The purpose of this study was to use endoscopically directed biopsies and scanning electron microscopy (SEM) to document the existence of Helicobacter pylori biofilms in human gastric mucosa. Patients underwent flexible esophagogastroduodenoscopies with three gastric mucosal biopsies. Rapid urease testing was performed to determine the presence or absence of H pylori. Urease positive and urease-negative control specimens were imaged with SEM to obtain detailed images of gastric mucosa for the identification of biofilm colonies. Samples were obtained from patients who underwent esophagogastroduodenoscopies. Eleven were found to be H pylori positive and nine were H pylori negative. These were imaged at 500x and 1000x with electron microscopy. Dense, mature biofilms were present and attached to the cell surface of H pylori-positive specimens and were absent in urease-negative controls. Photomicrographs were obtained. Biofilms are complex microbiological ecosystems where sessile bacteria surround themselves in a protective matrix. This lifestyle affords protection, allows for growth in hostile environments, and alters host physiology. Many have hypothesized that H pylori infections resulting in gastric ulcers may be a manifestation of biofilms. Our investigation is the first to photographically document the existence of H pylori biofilms on human gastric mucosa. This elucidation of the ecology and pathophysiology of the mucosa of the organism is important to our understanding of a potential mechanism of this organism's resistance to current therapy and how to better eradicate it in the future. PMID- 16713545 TI - Comparison of results of surgery in the upper third and more distal stomach. AB - The results of surgery for gastric cancer in the upper third of the stomach and the more distal region were compared to explain the comparatively poor outcomes in the former. Characteristics and therapeutic results of 1245 patients who underwent curative gastrectomy (244 with tumors in the upper third of the stomach and 1001 with more distal cancers) were compared. Survival was significantly lower in patients with cancer of the upper third of the stomach than in patients with more distal cancers. There was a significant difference between the two groups in the survival of patients with advanced gastric cancer, particularly in those with pN1. Among patients with tumors deeper than T2 and pN1, survival was significantly reduced in those aged or=50 but <100 ml/min (group 1; n = 77) and a group with Ccr values of >or=20 to <50 ml/min (group 2; n = 14). Preoperative patient characteristics, intraoperative parameters (including operation time and blood loss), and postoperative management and complications were evaluated. The preoperative evaluation showed no differences in liver function between the two groups, and there were no statistically significant differences between the two groups in intraoperative blood loss (522 ml in group 1 and 806 ml in group 2) or intraoperative urine volume (1.01 ml/kg per hour in group 1 and 0.75 ml/kg per hour in group 2). The difference between the two groups in postoperative complications was not statistically significant. None of the patients in group 2 required dialysis therapy, and no patients died as a result of hepatectomy or hepatectomy-related causes. Adequate indications, appropriate operative procedures, and perioperative management can enable hepatectomy to be performed safely in patients with nonuremic minimal renal failure. PMID- 16713549 TI - Abdominal lymphangiomas in adults. AB - Abdominal lymphangiomas are rare benign cystic tumors that can become locally invasive and often require resection. They arise in all ages and have a variable presentation. We performed a retrospective review of a single institution surgical experience with this lesion in adults. The pathology prospective database was reviewed to identify patients with surgically resected abdominal lymphangiomas from January 1986 to May 2004. Retrospective review and follow-up was performed for each patient. The six patients with abdominal lymphangiomas ranged in age from 38 to 66 years. They presented with a variety of signs and symptoms. All underwent CT scan that demonstrated a cystic lesion, but in only one third was the diagnosis made preoperatively. Tumors were located in the retroperitoneum, small bowel mesentery, liver, and pancreas. Five of the six tumors were completely resected. Two of the six required resection of adjacent or involved organs. Follow-up ranged between 6 months and 18 years. All had symptomatic relief after resection, and no patient showed evidence of recurrence in this time period. Abdominal lymphangiomas are rare. The correct diagnosis often remains elusive until tissue is obtained. The treatment of choice is complete surgical resection. When completely resected, these lesions seem not to recur, and the overall prognosis is excellent. PMID- 16713551 TI - Pneumatosis intestinalis and gas in the portal venous system. PMID- 16713552 TI - Persistent portal venous gas. AB - This case report describes a patient diagnosed with ongoing portal venous gas, initiated by a rather common Campylobacter enterocolitis and maintained by septic thrombophlebitis and possibly by chronic cholecystitis. Cholecystectomy attenuated the patient's septic condition. The etiology of portal venous gas determines both the patient's prognosis and the choice for either conservative or surgical treatment. This report describes persistence of portal venous gas for a long period and a possible role for chronic cholecystitis as a cause. PMID- 16713553 TI - Clearing a path through the patent thicket. AB - Patents do not always promote innovation, particularly when they restrict access to fundamental scientific discoveries and the tools of basic research. However, there are legal and policy approaches that may help to ameliorate problems associated with patenting these sorts of inventions. PMID- 16713554 TI - Astrocytes put down the broom and pick up the baton. AB - In this issue of Cell, Panatier et al. (2006) show that D-serine released by astrocytes, a type of glial cell in the brain, promotes NMDA receptor activity at synapses in the hypothalamus. These results confirm that glial cells participate in synaptic signaling and show that they can dictate the tenor of synaptic plasticity. PMID- 16713555 TI - DisA, a busy bee that monitors chromosome integrity. AB - When nutrients are limited many prokaryotic and some eukaryotic cells tuck their chromosomes safely away in resistant spores. However, before starting the sporulation process, the prokaryote Bacillus subtilis wisely ensures that its chromosome is intact. In this issue of Cell, Bejerano-Sagie et al. (2006) describe a protein, DisA, that is responsible for the surveillance of chromosome integrity during sporulation. PMID- 16713550 TI - Management of hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is one of the most common tumors globally, with varying prevalence based on endemic risk factors. In high-risk populations, including those with hepatitis B or C or with cirrhosis, serum alpha-fetoprotein (AFP) and screening ultrasound have improved detection of resectable HCC. Treatment options, including surgical resection, for patients with HCC must be selected based on the number and size of hepatic tumors, underlying hepatic function, patient condition, and available resources. An approach, which has been summarized shows the corresponding treatment choices under given clinical circumstances. For cirrhotic patients with less than three tumor nodules of a size less than 3 cm or a solitary HCC less than 5 cm, liver transplantation offers long-term survival similar to that observed in patients transplanted for nonmalignant disease. Ablative treatment using either chemical or thermal techniques provides locally effective tumor destruction. Transcatheter arterial chemoembolization (TACE) is commonly used for palliation of unresectable tumors as well as an adjunct to surgical resection, treatment of tumors before transplant, and in conjunction with other ablative therapies in a multimodality approach. Regional approaches to chemotherapy have produced more encouraging results than systemic chemotherapy, although both remain ineffective for long term tumor control. Several newer treatment modalities are under investigation, including gene therapy, tagged antibodies, isolated perfusion, and novel radiotherapy techniques. PMID- 16713556 TI - CYLD in ubiquitin signaling and tumor pathogenesis. AB - Absence of CYLD, which encodes a deubiquitinating enzyme, causes an inherited disease characterized by benign skin tumors. In this issue of Cell, Massoumi et al. (2006) show that CYLD deubiquitinates the coactivator Bcl-3, thereby preventing its translocation into the nucleus, where it normally interacts with NF-kappaB and activates transcription of proliferation genes in response to growth signals. PMID- 16713557 TI - The ataxia-ome: connecting disease proteins of the cerebellum. AB - In this issue of Cell, Lim et al. (2006) describe a protein-protein interaction network for inherited human ataxias, a group of diseases characterized by degeneration of cerebellar Purkinje cells. This protein interactome shows that the cerebellar ataxias not only share clinical and pathological characteristics but also have proteins, processes, and pathways in common. PMID- 16713558 TI - Using isoform-specific inhibitors to target lipid kinases. AB - In this issue of Cell, Knight et al. (2006) present a detailed pharmacological analysis of the PI3 kinase (PI3-K) family using small molecule inhibitors. They conclude that p110alpha, a PI3-K isoform, has a critical role in insulin signaling. In a related Cancer Cell paper, Fan et al. (2006) show that blocking activation of both p110alpha and the kinase mTOR with a small molecule inhibitor limits the growth of gliomas. PMID- 16713559 TI - The exosome and the proteasome: nano-compartments for degradation. AB - Exosomes and proteasomes are macromolecular complexes that posttranscriptionally regulate gene expression by degrading mRNAs and proteins, respectively. Although the two complexes act on different substrates and are composed of different subunits, they share a similar barrel-like architecture that appears to have evolved to restrict substrate access and prevent indiscriminate degradation. PMID- 16713560 TI - The timing of developmental transitions in plants. AB - Plants rely heavily on environmental cues to control the timing of developmental transitions. We are beginning to better understand what determines the timing of two of these transitions, the switch from juvenile to adult vegetative development and the transition to flowering. In this review, we discuss how RNA silencing mechanisms may influence the juvenile-to-adult vegetative switch. We also describe the discovery and regulation of a component of "florigen," the mobile flowering promotion signal that is involved in the transition to flowering. Parallel themes are beginning to emerge from a molecular comparison of these two developmental transitions. PMID- 16713561 TI - Cyld inhibits tumor cell proliferation by blocking Bcl-3-dependent NF-kappaB signaling. AB - Mutations in the CYLD gene cause tumors of hair-follicle keratinocytes. The CYLD gene encodes a deubiquitinase that removes lysine 63-linked ubiquitin chains from TRAF2 and inhibits p65/p50 NF-kappaB activation. Here we show that mice lacking Cyld are highly susceptible to chemically induced skin tumors. Cyld-/- tumors and keratinocytes treated with 12-O-tetradecanoylphorbol-13 acetate (TPA) or UV light are hyperproliferative and have elevated cyclin D1 levels. The cyclin D1 elevation is caused not by increased p65/p50 action but rather by increased nuclear activity of Bcl-3-associated NF-kappaB p50 and p52. In Cyld+/+ keratinocytes, TPA or UV light triggers the translocation of Cyld from the cytoplasm to the perinuclear region, where Cyld binds and deubiquitinates Bcl-3, thereby preventing nuclear accumulation of Bcl-3 and p50/Bcl-3- or p52/Bcl-3 dependent proliferation. These data indicate that, depending on the external signals, Cyld can negatively regulate different NF-kappaB pathways; inactivation of TRAF2 controls survival and inflammation, while inhibition of Bcl-3 controls proliferation and tumor growth. PMID- 16713562 TI - A checkpoint protein that scans the chromosome for damage at the start of sporulation in Bacillus subtilis. AB - In response to DNA damage, cells activate checkpoint signaling cascades to control cell-cycle progression and elicit DNA repair in order to maintain genomic integrity. The sensing and repair of lesions is critical for Bacillus subtilis cells entering the developmental process of sporulation as damaged DNA may prevent the cells from completing spore morphogenesis. We report the identification of the protein DisA (DNA integrity scanning protein, annotated YacK), which is required to delay the initiation of sporulation in response to chromosomal damage. DisA is a nonspecific DNA binding protein that forms a single focus, which moves rapidly within the bacterial cell, pausing at sites of DNA damage. We propose that the DisA focus scans along the chromosomes searching for lesions. Upon encountering a lesion, DisA delays entry into sporulation until the damage is repaired. PMID- 16713563 TI - Histone H2B monoubiquitination functions cooperatively with FACT to regulate elongation by RNA polymerase II. AB - Over the past years, a large number of histone posttranslational modifications have been described, some of which function to attain a repressed chromatin structure, while others facilitate activation by allowing access of regulators to DNA. Histone H2B monoubiquitination is a mark associated with transcriptional activity. Using a highly reconstituted chromatin-transcription system incorporating the inducible RARbeta2 promoter, we find that the establishment of H2B monoubiquitination by RNF20/40 and UbcH6 is dependent on the transcription elongation regulator complex PAF, the histone chaperone FACT, and transcription. H2B monoubiquitination facilitates FACT function, thereby stimulating transcript elongation and the generation of longer transcripts. These in vitro analyses and corroborating in vivo experiments demonstrate that elongation by RNA polymerase II through the nucleosomal barrier is minimally dependent upon (1) FACT and (2) the recruitment of PAF and the H2B monoubiquitination machinery. PMID- 16713564 TI - Downregulation of PP2A(Cdc55) phosphatase by separase initiates mitotic exit in budding yeast. AB - After anaphase, the high mitotic cyclin-dependent kinase (Cdk) activity is downregulated to promote exit from mitosis. To this end, in the budding yeast S. cerevisiae, the Cdk counteracting phosphatase Cdc14 is activated. In metaphase, Cdc14 is kept inactive in the nucleolus by its inhibitor Net1. During anaphase, Cdk- and Polo-dependent phosphorylation of Net1 is thought to release active Cdc14. How Net1 is phosphorylated specifically in anaphase, when mitotic kinase activity starts to decline, has remained unexplained. Here, we show that PP2A(Cdc55) phosphatase keeps Net1 underphosphorylated in metaphase. The sister chromatid-separating protease separase, activated at anaphase onset, interacts with and downregulates PP2A(Cdc55), thereby facilitating Cdk-dependent Net1 phosphorylation. PP2A(Cdc55) downregulation also promotes phosphorylation of Bfa1, contributing to activation of the "mitotic exit network" that sustains Cdc14 as Cdk activity declines. These findings allow us to present a new quantitative model for mitotic exit in budding yeast. PMID- 16713565 TI - Perception of the bacterial PAMP EF-Tu by the receptor EFR restricts Agrobacterium-mediated transformation. AB - Higher eukaryotes sense microbes through the perception of pathogen-associated molecular patterns (PAMPs). Arabidopsis plants detect a variety of PAMPs including conserved domains of bacterial flagellin and of bacterial EF-Tu. Here, we show that flagellin and EF-Tu activate a common set of signaling events and defense responses but without clear synergistic effects. Treatment with either PAMP results in increased binding sites for both PAMPs. We used this finding in a targeted reverse-genetic approach to identify a receptor kinase essential for EF Tu perception, which we called EFR. Nicotiana benthamiana, a plant unable to perceive EF-Tu, acquires EF-Tu binding sites and responsiveness upon transient expression of EFR. Arabidopsis efr mutants show enhanced susceptibility to the bacterium Agrobacterium tumefaciens, as revealed by a higher efficiency of T-DNA transformation. These results demonstrate that EFR is the EF-Tu receptor and that plant defense responses induced by PAMPs such as EF-Tu reduce transformation by Agrobacterium. PMID- 16713566 TI - Nontranscriptional regulation of SYK by the coactivator OCA-B is required at multiple stages of B cell development. AB - OCA-B was originally identified as a nuclear transcriptional coactivator that is essential for antigen-driven immune responses. The later identification of a membrane bound, myristoylated form of OCA-B suggested additional, unique functions in B cell signaling pathways. This study has shown that OCA-B also functions in the pre-B1-to-pre-B2 cell transition and, most surprisingly, that it directly interacts with SYK, a tyrosine kinase critical for pre-BCR and BCR signaling. This unprecedented type of interaction-a transcriptional coactivator with a signaling kinase-occurs in the cytoplasm and directly regulates SYK stability. This study indicates that OCA-B is required for pre-BCR and BCR signaling at multiple stages of B cell development through its nontranscriptional regulation of SYK. Combined with the deregulation of OCA-B target genes, this may help explain the multitude of defects observed in B cell development and immune responses of Oca-b-/- mice. PMID- 16713567 TI - Glia-derived D-serine controls NMDA receptor activity and synaptic memory. AB - The NMDA receptor is a key player in excitatory transmission and synaptic plasticity in the central nervous system. Its activation requires the binding of both glutamate and a co-agonist like D-serine to its glycine site. As D-serine is released exclusively by astrocytes, we studied the physiological impact of the glial environment on NMDA receptor-dependent activity and plasticity. To this end, we took advantage of the changing astrocytic ensheathing of neurons occurring in the supraoptic nucleus during lactation. We provide direct evidence that in this hypothalamic structure the endogenous co-agonist of NMDA receptors is D-serine and not glycine. Consequently, the degree of astrocytic coverage of neurons governs the level of glycine site occupancy on the NMDA receptor, thereby affecting their availability for activation and thus the activity dependence of long-term synaptic changes. Such a contribution of astrocytes to synaptic metaplasticity fuels the emerging concept that astrocytes are dynamic partners of brain signaling. PMID- 16713568 TI - Miniature neurotransmission stabilizes synaptic function via tonic suppression of local dendritic protein synthesis. AB - Activity deprivation in neurons induces a slow compensatory scaling up of synaptic strength, reflecting a homeostatic mechanism for stabilizing neuronal activity. Prior studies have focused on the loss of action potential (AP) driven neurotransmission in synaptic homeostasis. Here, we show that the miniature synaptic transmission that persists during AP blockade profoundly shapes the time course and mechanism of homeostatic scaling. A brief blockade of NMDA receptor (NMDAR) mediated miniature synaptic events ("minis") rapidly scales up synaptic strength, over an order of magnitude faster than with AP blockade alone. The rapid scaling induced by NMDAR mini blockade is mediated by increased synaptic expression of surface GluR1 and the transient incorporation of Ca2+-permeable AMPA receptors at synapses; both of these changes are implemented locally within dendrites and require dendritic protein synthesis. These results indicate that NMDAR signaling during miniature synaptic transmission serves to stabilize synaptic function through active suppression of dendritic protein synthesis. PMID- 16713569 TI - A protein-protein interaction network for human inherited ataxias and disorders of Purkinje cell degeneration. AB - Many human inherited neurodegenerative disorders are characterized by loss of balance due to cerebellar Purkinje cell (PC) degeneration. Although the disease causing mutations have been identified for a number of these disorders, the normal functions of the proteins involved remain, in many cases, unknown. To gain insight into the function of proteins involved in PC degeneration, we developed an interaction network for 54 proteins involved in 23 inherited ataxias and expanded the network by incorporating literature-curated and evolutionarily conserved interactions. We identified 770 mostly novel protein-protein interactions using a stringent yeast two-hybrid screen; of 75 pairs tested, 83% of the interactions were verified in mammalian cells. Many ataxia-causing proteins share interacting partners, a subset of which have been found to modify neurodegeneration in animal models. This interactome thus provides a tool for understanding pathogenic mechanisms common for this class of neurodegenerative disorders and for identifying candidate genes for inherited ataxias. PMID- 16713570 TI - Signaling from across the way: transactivation of VEGF receptors by HSPGs. AB - Heparan sulfate proteoglycans (HSPGs) influence the signaling of many growth factors, and recent work shows that they can promote VEGF signaling in trans from the surface of an adjacent cell. This mode of signaling also alters the activation state and internalization of VEGF receptors. PMID- 16713571 TI - A confederacy of kinases: Cdk2 and Cdk4 conspire to control embryonic cell proliferation. AB - The mouse embryo is surprisingly resistant to loss of individual cyclins or cyclin-dependent kinases. In a recent issue of Developmental Cell, Berthet et al. (2006) describe collaboration of Cdk2 and Cdk4 in embryogenesis that is revealed only upon their simultaneous loss, resulting in inappropriate activation of the retinoblastoma protein and embryonic lethality. PMID- 16713572 TI - If the prophet does not come to the mountain: dynamics of signaling complexes in NF-kappaB activation. AB - Recruitment of the NF-kappaB-activating IKK signaling complex to the TNF receptor is shown to be driven by induced binding of NEMO, a regulatory component of this complex, to K63-linked polyubiquitin chains attached to RIP1, a receptor associated adaptor protein (Ea et al., 2006 [in a recent issue of Molecular Cell]; Li et al., 2006; Wu et al., 2006a). PMID- 16713573 TI - Autophagy by ARF: a short story. AB - In this issue of Molecular Cell, Reef et al. (2006) describe a shortened unstable form of the ARF tumor suppressor protein that localizes within mitochondria, where it reduces membrane potential and triggers autophagy. Could this account for the Mdm2- and p53-independent tumor suppressive effects of ARF? PMID- 16713574 TI - VHL and p53: tumor suppressors team up to prevent cancer. AB - In a recent issue of Molecular Cell, Roe et al. (2006) report that the von Hippel Lindau (VHL) protein is a positive regulator of p53, thus providing insight into the mechanisms by which VHL loss of function leads to cancer. PMID- 16713575 TI - In vivo molecular evolution reveals biophysical origins of organismal fitness. AB - In nature, evolution occurs through the continuous adaptation of a population to its environment. At the molecular level, adaptive changes in protein sequence and expression impact organismal fitness and, consequently, dictate population dynamics. Here, we have used a "weak link" method to favor variations in one gene, allowing adaptation to thermostability to be studied in molecular detail as bacteria were grown continuously for approximately 1500 generations. Surprisingly, only six mutant alleles, representing less than 1% of the possible missense mutations, were observed, suggesting a highly constrained molecular landscape during protein evolution. The changes in organismal fitness were linked directly to incremental increases in enzyme stability and activity maxima and corresponded to the narrow temperature ranges where each mutant enjoyed success within the overall population. Thus, continuous evolution of a single gene permits a quantitative approach that extends from the phenotypes of the microbial populations to their underlying biophysical basis. PMID- 16713576 TI - Central pore residues mediate the p97/VCP activity required for ERAD. AB - The AAA-ATPase p97/VCP facilitates protein dislocation during endoplasmic reticulum-associated degradation (ERAD). To understand how p97/VCP accomplishes dislocation, a series of point mutants was made to disrupt distinguishing structural features of its central pore. Mutants were evaluated in vitro for ATPase activity in the presence and absence of synaptotagmin I (SytI) and in vivo for ability to process the ERAD substrate TCRalpha. Synaptotagmin induces a 4 fold increase in the ATPase activity of wild-type p97/VCP (p97/VCP(wt)), but not in mutants that showed an ERAD impairment. Mass spectrometry of crosslinked synaptotagmin . p97/VCP revealed interactions near Trp551 and Phe552. Additionally, His317, Arg586, and Arg599 were found to be essential for substrate interaction and ERAD. Except His317, which serves as an interaction nexus, these residues all lie on prominent loops within the D2 pore. These data support a model of substrate dislocation facilitated by interactions with p97/VCP's D2 pore. PMID- 16713577 TI - A short mitochondrial form of p19ARF induces autophagy and caspase-independent cell death. AB - The tumor suppressor functions of p19(ARF) have been attributed to its ability to induce cell cycle arrest or apoptosis by activating p53 and regulating ribosome biogenesis. Here we describe another cellular function of p19(ARF), involving a short isoform (smARF, short mitochondrial ARF) that localizes to a Proteinase K resistant compartment of the mitochondria. smARF is a product of internal initiation of translation at Met45, which lacks the nucleolar functional domains. The human p14(ARF) mRNA likewise produces a shorter isoform. smARF is maintained at low levels via proteasome-mediated degradation, but it increases in response to viral and cellular oncogenes. Ectopic expression of smARF reduces mitochondrial membrane potential (DeltaPsim) without causing cytochrome c release or caspase activation. The dissipation of DeltaPsim does not depend on p53 or Bcl 2 family members. smARF induces massive autophagy and caspase-independent cell death that can be partially rescued by knocking down ATG5 or Beclin-1, suggesting a different prodeath function for this short isoform. PMID- 16713578 TI - PIASxalpha differentially regulates the amplitudes of transcriptional responses following activation of the ERK and p38 MAPK pathways. AB - Activation of the MAP kinase pathways leads to changes in gene expression profiles through direct targeting of transcription factors and their coregulators. Here we identify PIASxalpha as a key regulator that determines the differential response of the transcription factor Elk-1 to the ERK and the stress activated p38 MAP kinase pathways. While PIASxalpha functions as a coactivator to facilitate SUMO and HDAC-2 removal from Elk-1 in response to ERK pathway activation, PIASxalpha acts in the opposite manner to inhibit HDAC-2 and SUMO loss following stress-activated MAP kinase pathway signaling. Thus, PIASxalpha either enhances or dampens down the activation of Elk-1 target genes, depending on the pathway activated. p38 MAP kinase-mediated PIASxalpha phosphorylation allows it to switch between these two alternative modes of operation. Thus, PIASxalpha acts as a key signal integrator that permits different responses from the same transcription factor, depending on the signaling pathway that is activated. PMID- 16713579 TI - A kinase-independent function of c-Abl in promoting proteolytic destruction of damaged DNA binding proteins. AB - Damaged DNA binding proteins (DDBs) play a critical role in the initial recognition of UV-damaged DNA and mediate recruitment of nucleotide excision repair factors. Previous studies identified DDB2 as a target of the CUL-4A ubiquitin ligase. However, the biochemical mechanism governing DDB proteolysis and its underlying physiological function in the removal of UV-induced DNA damage are largely unknown. Here, we report that the c-Abl nonreceptor tyrosine kinase negatively regulates the repair of UV-induced photolesions on genomic DNA. Biochemical studies revealed that c-Abl promotes CUL-4A-mediated DDB ubiquitination and degradation in a manner that does not require its tyrosine kinase activity both under normal growth conditions and following UV irradiation. Moreover, c-Abl activates DDB degradation in part by alleviating the inhibitory effect of CAND1/TIP120A on CUL-4A. These results revealed a kinase-independent function of c-Abl in a ubiquitin-proteolytic pathway that regulates the damage recognition step of nucleotide excision repair. PMID- 16713581 TI - Three-dimensional structure of the human DNA-PKcs/Ku70/Ku80 complex assembled on DNA and its implications for DNA DSB repair. AB - DNA-PKcs is a large (approximately 470 kDa) kinase that plays an essential role in the repair of DNA double-strand breaks (DSBs) by nonhomologous end joining (NHEJ). DNA-PKcs is recruited to DSBs by the Ku70/Ku80 heterodimer, with which it forms the core of a multiprotein complex that promotes synapsis of the broken DNA ends. We have purified the human DNA-PKcs/Ku70/Ku80 holoenzyme assembled on a DNA molecule. Its three-dimensional (3D) structure at approximately 25 Angstroms resolution was determined by single-particle electron microscopy. Binding of Ku and DNA elicits conformational changes in the FAT and FATC domains of DNA-PKcs. Dimeric particles are observed in which two DNA-PKcs/Ku70/Ku80 holoenzymes interact through the N-terminal HEAT repeats. The proximity of the dimer contacts to the likely positions of the DNA ends suggests that these represent synaptic complexes that maintain broken DNA ends in proximity and provide a platform for access of the various enzymes required for end processing and ligation. PMID- 16713580 TI - ATR kinase activation mediated by MutSalpha and MutLalpha in response to cytotoxic O6-methylguanine adducts. AB - S(N)1-type alkylating agents that produce cytotoxic O(6)-methyl-G (O(6)-meG) DNA adducts induce cell cycle arrest and apoptosis in a manner requiring the DNA mismatch repair (MMR) proteins MutSalpha and MutLalpha. Here, we show that checkpoint signaling in response to DNA methylation occurs during S phase and requires DNA replication that gives rise to O(6)-meG/T mispairs. DNA binding studies reveal that MutSalpha specifically recognizes O(6)-meG/T mispairs, but not O(6)-meG/C. In an in vitro assay, ATR-ATRIP, but not RPA, is preferentially recruited to O(6)-meG/T mismatches in a MutSalpha- and MutLalpha-dependent manner. Furthermore, ATR kinase is activated to phosphorylate Chk1 in the presence of O(6)-meG/T mispairs and MMR proteins. These results suggest that MMR proteins can act as direct sensors of methylation damage and help recruit ATR ATRIP to sites of cytotoxic O(6)-meG adducts to initiate ATR checkpoint signaling. PMID- 16713582 TI - Transcription impairment and cell migration defects in elongator-depleted cells: implication for familial dysautonomia. AB - Mutations in IKBKAP, encoding a subunit of Elongator, cause familial dysautonomia (FD), a severe neurodevelopmental disease with complex clinical characteristics. Elongator was previously linked not only with transcriptional elongation and histone acetylation but also with other cellular processes. Here, we used RNA interference (RNAi) and fibroblasts from FD patients to identify Elongator target genes and study the role of Elongator in transcription. Strikingly, whereas Elongator is recruited to both target and nontarget genes, only target genes display histone H3 hypoacetylation and progressively lower RNAPII density through the coding region in FD cells. Interestingly, several target genes encode proteins implicated in cell motility. Indeed, characterization of IKAP/hELP1 RNAi cells, FD fibroblasts, and neuronal cell-derived cells uncovered defects in this cellular function upon Elongator depletion. These results indicate that defects in Elongator function affect transcriptional elongation of several genes and that the ensuing cell motility deficiencies may underlie the neuropathology of FD patients. PMID- 16713583 TI - Elongation arrest by SecM via a cascade of ribosomal RNA rearrangements. AB - In E. coli, the SecM nascent polypeptide causes elongation arrest, while interacting with 23S RNA bases A2058 and A749-753 in the exit tunnel of the large ribosomal subunit. We compared atomic models fitted by real-space refinement into cryo-electron microscopy reconstructions of a pretranslocational and SecM-stalled E. coli ribosome complex. A cascade of RNA rearrangements propagates from the exit tunnel throughout the large subunit, affecting intersubunit bridges and tRNA positions, which in turn reorient small subunit RNA elements. Elongation arrest could result from the inhibition of mRNA.(tRNAs) translocation, E site tRNA egress, and perhaps translation factor activation at the GTPase-associated center. Our study suggests that the specific secondary and tertiary arrangement of ribosomal RNA provides the basis for internal signal transduction within the ribosome. Thus, the ribosome may itself have the ability to regulate its progression through translation by modulating its structure and consequently its receptivity to activation by cofactors. PMID- 16713584 TI - Genetically encoded but nonpolypeptide prolyl-tRNA functions in the A site for SecM-mediated ribosomal stall. AB - The arrest sequence, FXXXXWIXXXXGIRAGP, of E. coli SecM interacts with the ribosomal exit tunnel, thereby interfering with translation elongation. Here, we studied this elongation arrest in vitro using purified translation components. While a simplest scenario would be that elongation is arrested beyond Pro166, the last arrest-essential amino acid, and that the Pro166 codon is positioned at the P site of the ribosomal peptidyl transferase center (PTC), our toeprint analyses revealed that the ribosome actually stalls when the Pro166 codon is positioned at the A site. Northern hybridization identification of the polypeptide bound tRNA and mass determination showed that the last amino acid of the arrested peptidyl tRNA is Gly165, which is only inefficiently transferred to Pro166. Also, puromycin does not effectively release the arrested peptidyl-tRNA under the conditions of A site occupancy by Pro166-tRNA. These results reveal that secM encoded Pro166-tRNA functions as a nonpolypeptide element in fulfilling SecM's role as a secretion monitor. PMID- 16713585 TI - Recapitulation of short RNA-directed translational gene silencing in vitro. AB - microRNAs (miRNAs) are a large class of endogenous short RNAs that repress gene expression. Many miRNAs are conserved throughout evolution, and dysregulation of miRNA pathways has been correlated with an increasing number of human diseases. In animals, miRNAs typically bind to the 3' untranslated region (3'UTR) of target mRNAs with imperfect sequence complementarity and repress translation. Despite their importance in regulating biological processes in numerous organisms, the mechanisms of miRNA function are largely unknown. Here, we report in vitro reactions for miRNA-directed translational gene silencing. These reactions faithfully recapitulate known in vivo hallmarks of mammalian miRNA function, including a requirement for a 5' phosphate and perfect complementarity to the mRNA target in the 5' seed region. Translational gene silencing by miRNAs in vitro requires target mRNAs to possess a 7-methyl G cap and a polyA tail, whereas increasing polyA tail length alone can increase miRNA silencing activity. PMID- 16713587 TI - Mood alters amygdala activation to sad distractors during an attentional task. AB - BACKGROUND: A behavioral hallmark of mood disorders is biased perception and memory for sad events. The amygdala is poised to mediate internal mood and external event processing because of its connections with both the internal milieu and the sensory world. There is little evidence showing that the amygdala's response to sad sensory stimuli is functionally modulated by mood state, however. METHODS: We investigated the impact of mood on amygdala activation evoked by sad and neutral pictures presented as distractors during an attentional oddball task. Healthy adults underwent functional magnetic resonance imaging during task runs that were preceded by sad or happy movie clips. Happy and sad mood induction was conducted within-subjects on consecutive days in counterbalanced order. RESULTS: Amygdala activation to sad distractors was enhanced after viewing sad movies relative to happy ones and was correlated with reaction time costs to detect attentional targets. The activation was higher in female subjects in the right hemisphere. The anterior cingulate, ventromedial and orbital prefrontal cortex, insula, and other posterior regions also showed enhanced responses to sad distractors during sad mood. CONCLUSIONS: These findings reveal brain mechanisms that integrate emotional input and current mood state, with implications for understanding cognitive distractibility in depression. PMID- 16713586 TI - Association of specific haplotypes of neurotrophic tyrosine kinase receptor 2 gene (NTRK2) with vulnerability to nicotine dependence in African-Americans and European-Americans. AB - BACKGROUND: The gene encoding neurotrophic tyrosine kinase receptor 2 (NTRK2) has been localized to a region on chromosome 9q22-q23 that showed a "suggestive" linkage to nicotine dependence (ND) in our previous linkage analyses. However, no association of NTRK2 with ND has been identified. METHODS: Family-based association analyses of 2037 participants (1366 African Americans [AA], 671 European Americans [EA]) representing 602 nuclear families were performed to evaluate association of nine single nucleotide polymorphisms (SNPs) within NTRK2 with ND. RESULTS: Individual SNP-based association analysis indicated that in the EA sample, SNPs rs1659400 and rs1187272 were significantly associated with at least one adjusted ND measure. Haplotype analysis revealed that even after Bonferroni correction, the haplotype T-T-A of rs1659400-rs1187272-rs1122530 had a highly significant positive association, with adjusted ND measures in the EA sample (max Z = 3.78; p = .0001, frequency 59.9%). We further identified a major haplotype, T-G-C-A-A (26%), formed by rs993315-rs736744-rs920776-rs4075274 rs729560, which showed a significant positive association (max Z = 2.97, p = .003) with adjusted ND measures in the AA sample. CONCLUSIONS: These results strongly suggest that NTRK2 is a susceptibility gene for ND. These findings imply that NTRK2 plays a role in the etiology of ND and represents an important biological candidate for further investigation. PMID- 16713588 TI - Lateralization of unimanual and bimanual motor imagery. AB - Most studies of motor imagery have examined motor cortex function during imagery of dominant hand movement. The aim of this study was to examine the modulation of excitability in the dominant and non-dominant corticomotor pathways during kinesthetic motor imagery of unimanual and bimanual movement. Transcranial magnetic stimulation (TMS) was applied over the contralateral motor cortex (M1) to elicit motor-evoked potentials (MEPs) in the abductor pollicis brevis (APB) and abductor digiti minimi (ADM) muscles of each hand, in two separate sessions. Transcutaneous electrical stimuli were also delivered to the median nerve at each wrist, to elicit F-waves from APB. Fifteen right-handed volunteers imagined unimanual and bimanual phasic thumb movements, paced with a 1-Hz auditory metronome. Stimuli were delivered at rest, and either 50 ms before (ON phase), or 450 ms after (OFF phase), the metronome beeps. Significant MEP amplitude facilitation occurred only in right APB, during the ON phase of motor imagery of the right hand and both hands. Significant temporal modulation of right APB MEP amplitude was observed during motor imagery of right, left and bimanual performance. F-wave persistence and amplitude were unaffected by imagery. These results demonstrate that the motor imagery is lateralized to the left (dominant) hemisphere, which is engaged by imagery of each hand separately, and bimanual imagery. This finding has implications for the use of motor imagery in rehabilitation. PMID- 16713589 TI - CSF serotonin, 5-hydroxyindolacetic acid and neuropeptide Y levels in severe major depressive disorder. AB - Serotonin (5-HT) and 5-hydroxyindolacetic acid (5-HIAA) have been the putative markers of MDD. Neuropeptide Y (NPY) may have an important role in the pathophysiology of major depressive disorder (MDD). However, direct measures of cerebrospinal fluid (CSF) 5-HT and NPY in severe MDD have been lacking. In the present study, we examined CSF 5-HT, 5-HIAA and NPY levels and correlate them with gender and suicidal behavior of severe major depressive disorder. Forty drug free subjects with a severe major depressive disorder and forty control subjects underwent lumber puncture and psychiatric evaluation. Cerebrospinal fluid levels of 5-HT, 5-HIAA and NPY were assayed by enzyme-linked immunosorbent assay (ELISA) test. The relationships among 5-HT, 5-HIAA, NPY and clinical variables were statistically evaluated. There were no differences between severe major depressive disorder and controls in all parameters measured. In severe MDD group, significantly lower CSF 5-HT and higher 5-HT turnover (5-HIAA/5-HT) were found in female patients compared with male patients. The patients with intense suicidal intents and suicidal attempts had significantly lower CSF 5-HT compared to patients with nonsuicidal intents. Additionally, significantly lower CSF NPY was found in first episode depressed patients compared with recurrent depressed patients. Gender-related difference in CSF 5-HT implied a female preponderance in major depression to some extent. Cerebrospinal fluid 5-HT levels and 5-HT turnover (5-HIAA/5-HT) could be valuable tools for prediction of suicidality and potential markers for evaluating major depressive disorder. NPY was perhaps a marker for first episode depression. PMID- 16713590 TI - Enhanced accumulation of tau in doubly transgenic mice expressing mutant betaAPP and presenilin-1. AB - Abeta amyloidosis and tauopathy are characteristic changes in the brain of Alzheimer's disease. Although much evidence suggests that Abeta deposit is a critical initiation factor, the pathological pathway between Abeta amyloidosis and tau accumulation remains unclear. Tau accumulation was examined in the doubly transgenic mouse (APP-PS) expressing betaAPP(KM670/671NL) (Tg2576) and presenilin 1 L286V (PS-1 L286Vtg). Accelerated and enhanced Abeta amyloid deposits were detected from 8 weeks. Tau accumulation appeared at 4.5 months and markedly increased in dystrophic neurites around Abeta amyloid. Accumulated tau was phosphorylated, conformationally altered, and argyrophilic. Expression of tau and accumulation of sarkosyl-insoluble phosphorylated tau were increased in APP-PS brains compared with those of Tg2576 mice. Straight or twisted tubules mimicking paired helical filament were revealed at electron microscopic level in 16-month old APP-PS. These findings suggest that mutant presenilin-1 accelerated Abeta induced tauopathy and further promoted fibril formation of tau. PMID- 16713592 TI - Comparison of the results for three automated immunoassay systems in determining serum HBV markers. AB - BACKGROUND: Automated immunoassay analyzers are used to identify hepatitis B virus (HBV) serum markers. In regions with high prevalence of HBV, it is imperative to compare test results from different immunoassay analyzers. METHODS: Samples from 496 subjects were collected and HBV markers were determined (double blind, parallel manner) using Abbott AxSYM, Roche Modular Analytics E170, and Abbott Architect i2000). RESULTS: Concurrence between AxSYM and E170 was 97.78% for HBsAg, 91.13% for anti-HBs, 98.79% for anti-HBc, 98.39% for HBeAg, and 88.91% for anti-HBe. Positive rates of anti-HBs and anti-HBe from AxSYM were lower than E170 (P<0.01). Concurrence between AxSYM and Architect i2000 was 98.79% for HBsAg, 91.33% for anti-HBs, 95.97% for anti-HBc, 98.39% for HBeAg, and 95.77% for anti-HBe. Positive anti-HBs rates from AxSYM were lower than Architect i2000 (P<0.01). Concurrence between E170 and Architect i2000 was 97.38% for HBsAg, 94.15% for anti-HBs, 95.56% for anti-HBc, 99.60% for HBeAg, and 88.10% for anti HBe. Positive anti-HBe rates using Architect i2000 were lower than E170 (P<0.01). Overall, the greatest differences were observed in samples with low-level serum HBV markers. CONCLUSION: Significant discrepancies were observed among results for the 3 automated immunoassay analyzers, especially for low-level anti-HBs and anti-HBe results. PMID- 16713591 TI - Carbon monoxide released by CORM-3 inhibits human platelets by a mechanism independent of soluble guanylate cyclase. AB - OBJECTIVE: Carbon monoxide (CO) modulates several physiological functions through activation of a cGMP-dependent pathway similar to that of nitric oxide (NO). Here we investigated the possible involvement of soluble guanylate cyclase in the anti aggregatory effect of micromolar concentrations of CO released by a novel, water soluble, CO releasing molecule (CORM) in human platelets. METHODS: Human platelet aggregation was induced by collagen or thrombin, and the effects of CO releasing molecule (CORM-3) and an NO donor on platelet aggregation were compared. RESULTS: CORM-3 liberated CO in a time- and concentration-dependent manner as evidenced by the formation of carbon monoxy myoglobin (MbCO) using a spectrophotometric assay. When added to washed platelets, CORM-3 (10-300 microM) inhibited collagen- and thrombin-induced aggregation in a concentration-dependent manner. The anti aggregatory effect of CORM-3 was reversed by deoxy-Mb (50 microM). Interestingly, in the presence of an inhibitor of guanylate cyclase (ODQ, 5 microM), inhibition of collagen-induced aggregation by CORM-3 was not blocked but potentiated. Under the same experimental conditions, inhibition of platelet aggregation by an NO donor (SNAP, 1 microM) was prevented by ODQ. In collagen-induced or thrombin induced platelet aggregation, a stimulator of guanylate cyclase (YC-1, 0.3 microM) did not alter the effect of CORM-3, whereas it markedly potentiated the inhibition of platelet aggregation mediated by SNAP. Notably, CORM-3-induced inhibition of platelet aggregation was of similar degree when platelets were activated by a low (20 mU/ml) or by high concentration of thrombin (100-200 mU/ml), whereas NO donors (SNP and SNAP)- or carbaprostacylin (cPGI(2))-induced effects were considerably attenuated when platelets were activated by high concentrations of thrombin. CONCLUSIONS: Inhibition of platelet aggregation by CO released by a novel, water-soluble CORM is not mediated by activation of soluble guanylate cyclase. In contrast to NO and PGI(2), CO effectively inhibits platelets even when cells are activated excessively. We suggest that despite the fact that CO is not a potent inhibitor of platelet activation, it may gain importance when NO and PGI(2) alone are insufficient to overcome excessive platelet activation. PMID- 16713593 TI - Tissue factor and tissue factor pathway inhibitor: a potential role in pregnancy and obstetric vascular complications? AB - Tissue factor (TF) is the principal cellular initiator of normal blood coagulation. As a result it is considered to be a major regulator of haemostasis and thrombogenesis. In vivo TF activity is regulated by specific circulating inhibitor known as "tissue factor pathway inhibitor (TFPI)". TF is also essential for other cellular processes including embryogenesis and angiogenesis as well as in implantation where it is particularly important in the first trimester. TF is highly expressed in syncytiotrophoblasts (STB) while TFPI is expressed in human umbilical vein endothelial cells (HUVEC). TFPI may be internalized via an endocytic pathway and recycled to the cell surface. The procoagulant tendency of STB may reflect a physiological need for immediate inhibition of hemorrhage in the placental intervillous spaces. Furthermore, the haemostatic balance involving STB and HUVEC may be critical for normal placental function and pregnancy outcome. Homozygous knockouts of both TF and TFPI are generally lethal in fetal mice; heterozygotes survive but with altered coagulation parameters. Despite their apparent association with placental microcirculation-thrombi-formation only few studies have addressed the role of TF and TFPI in the pathogenesis of gestational vascular complications. In this context, detailed studies could provide clinically relevant information. PMID- 16713594 TI - Evaluation of cardiac laboratory markers in patients with systemic sclerosis. AB - OBJECTIVES: Myocardial involvement is frequent in systemic sclerosis, but symptoms are usually delayed and non-specific, thus often misrecognized. The aim of this study was the evaluation of the early subclinical cardiac involvement in patients with systemic sclerosis by means of non-invasive laboratory cardiac markers. DESIGN AND METHODS: Cardiac troponin T (cTnT), ischemia modified albumin (IMA) and NT-prohormone-brain natriuretic peptide (NT-proBNP) were measured in 40 female patients with systemic sclerosis and in 40 matched healthy controls. RESULTS: Patients with systemic sclerosis displayed significantly increased concentrations of serum IMA (106 versus 93.5 kunits/l, P < 0.0001) and NT-proBNP (89 versus 37 pg/ml, P < 0.0001), whereas no significant differences could be observed in both IMA and NT-proBNP values in limited versus diffuse pattern of disease. CONCLUSIONS: The increased levels of NT-proBNP and IMA could be considered a sign of early myocardial involvement, warranting further heart examination and a regular follow-up. PMID- 16713595 TI - Albumin locust valley: a new case of analbuminemia. AB - OBJECTIVE: To discover and document cases of the rare disease analbuminemia. DESIGN AND METHODS: Blood specimens of a subject were analyzed by routine clinical laboratory procedures, by immunochemical tests for albumin, and by electrophoresis with immunofixation using anti-human serum albumin (HSA). Single strand conformational polymorphism (SSCP), heteroduplex analysis (HA), and DNA sequencing of the 14 exons of the HSA gene were conducted on DNA from leukocytes. RESULTS: Albumin concentration was 0.003 g/L; serum globulins and cholesterol were elevated. Immunoelectrophoresis showed no trace of albumin in any of the serum components. The coding region plus all mRNA splice sites were normal and mutation-free, and SSCP and HA showed no abnormalities. CONCLUSIONS: Data define a bona fide case of analbuminemia. We suggest that the mutation causing the analbuminemic trait in this subject might involve a remote regulatory element. PMID- 16713596 TI - Differing effects of dexamethasone and diclofenac on posterior capsule opacification-like changes in a rat lens explant model. AB - Posterior capsular opacification (PCO) arises from lens cells that remain associated with the lens capsule after cataract surgery and subsequently become abnormal, proliferate and migrate into the visual pathway. In this study, a rat lens explant model was used to assess the effects of the prototype steroidal and non-steroidal anti-inflammatory drugs, dexamethasone (DEX) and diclofenac (DIC), on epithelial cells undergoing PCO-like changes. Such drugs are widely used at the time of cataract surgery. TGFbeta2 and FGF-2 were added sequentially and explants were cultured for up to 30 days, with or without addition of DEX or DIC at a clinically relevant concentration. Without DEX or DIC, explants became multilayered and cells tended to retract into PCO-like plaques. Inclusion of DEX, but not DIC, resulted in transient formation of needle-like cells, enhanced cell coverage, and the retention a monolayer of migratory cells surrounding PCO-like plaques. With or without drug addition, most cells became aberrant, as indicated by loss of Pax6 expression and the presence of PCO markers alpha-smooth muscle actin and type I collagen; however, DEX and DIC both strongly enhanced type I collagen accumulation. Furthermore, DEX enhanced cell coverage in explants treated with TGFbeta alone. Thus the behaviour of lens cells was significantly and differentially affected by the presence of DEX and DIC, highlighting the possibility that drugs used to control inflammation after cataract surgery, and the clinician's choice of drugs, may influence PCO development. PMID- 16713598 TI - Cord blood rescues stroke-induced changes in splenocyte phenotype and function. AB - The neuroprotective mechanism of human umbilical cord blood cells (HUCBC) in the rat middle cerebral artery occlusion (MCAO) stroke model remains uncertain. Given the inflammatory sequelae that occur following stroke, we investigated whether HUCBC protection could be derived from the modulation of this immuno-inflammatory event, suggested by the attraction of the HUCBC to the spleen. We found that, following MCAO, rat spleen size was reduced concomitantly with their CD8+ T-cell counts. Interestingly, MCAO-induced spleen size reduction correlated with the extent of ischemic damage, however, HUCBC treatment rescued the spleen weight, splenic CD8+ T-cell counts, as well as the amount of brain injury. Additionally, splenocyte proliferation assays demonstrated that HUCBC treatment opposed MCAO associated T-cell proliferation by increasing the production of IL-10 while decreasing IFN-gamma. Taken together, these results suggest a novel immunomodulatory mechanism by which HUCBC mediate protection in the rat MCAO model of stroke. PMID- 16713599 TI - Rsp5 regulates expression of stress proteins via post-translational modification of Hsf1 and Msn4 in Saccharomyces cerevisiae. AB - Rsp5 is an essential E3 ubiquitin ligase in Saccharomyces cerevisiae and is known to ubiquitinate plasma membrane permeases followed by endocytosis and vacuolar degradation. We previously isolated the rsp5 mutant that is hypersensitive to various stresses, suggesting that Rsp5 is involved in degradation of stress induced abnormal proteins. Here, we analyzed the ability to refold the proteins by stress proteins in the rsp5 mutant. The transcription of stress protein genes in the rsp5 mutant was significantly lower than that in the wild-type strain when exposed to temperature up-shift, ethanol or sorbitol. Interestingly, the amounts of transcription factors Hsf1 and Msn4 were remarkably defective in the rsp5 mutant. These results suggest that expression of stress proteins are mediated by Rsp5 and that Rsp5 primarily regulates post-translational modification of Hsf1 and Msn4. PMID- 16713597 TI - Vascular endothelial growth factor receptors: molecular mechanisms of activation and therapeutic potentials. AB - Angiogenesis-associated eye diseases are among the most common cause of blindness in the United States and worldwide. Recent advances in the development of angiogenesis-based therapies for treatment of angiogenesis-associated diseases have provided new hope in a wide variety of human diseases ranging from eye diseases to cancer. One group of growth factor receptors critically implicated in angiogenesis is vascular endothelial growth factor receptors (VEGFR), a subfamily of receptor tyrosine kinases (RTKs). VEGFR-1 and VEGFR-2 are closely related receptor tyrosine kinases and have both common and specific ligands. VEGFR-1 is a kinase-impaired RTK and its kinase activity is suppressed by a single amino acid substitution in its kinase domain and by its carboxyl terminus. VEGFR-2 is highly active kinase, stimulates a variety of signaling pathways and broad biological responses in endothelial cells. The mechanisms that govern VEGFR-2 activation, its ability to recruit signaling proteins and to undergo downregulation are highly regulated by phosphorylation activation loop tyrosines and its carboxyl terminus. Despite their differential potentials to undergo tyrosine phosphorylation and kinase activation, both VEGFR-1 and VEGFR-2 are required for normal embryonic development and pathological angiogenesis. VEGFR-1 regulates angiogenesis by mechanisms that involve ligand trapping, receptor homodimerization and heterodimerization. This review highlights recent insights into the mechanism of activation of VEGFR-1 and VEGFR-2, and focuses on the signaling pathways employed by VEGFR-1 and VEGFR-2 that regulate angiogenesis and their therapeutic potentials in angiogenesis-associated diseases. PMID- 16713600 TI - Topology of protease activities reflects atherothrombotic plaque complexity. AB - The pathological remodeling of the arterial wall in atherosclerosis involves protease activities, which play a major role in complications, through plaque rupture. Here, we investigated the release of active proteases by human carotid plaques in relation to (1) the degree of lesion complexity and (2) their compartmentalization between cap, core and media. Eighty human carotid endarterectomy specimens were dissected into culprit stenosing (CPs) and adjacent non-complicated/non-stenosing plaques (NPs). Thirty-five additional CPs were microdissected into cap, core and media. All specimens were compared to control non-atherosclerotic endarteries for the release of components of the plasminogen/plasmin system and matrix metalloproteinases (MMPs). Results show a greater release of the plasminogen activators (PAs), plasmin and active MMPs by CPs compared to NPs, whereas healthy arteries released even lower levels. Furthermore, we highlight a functional interaction between these proteases in human atherosclerotic tissues and more importantly, we demonstrate that the core constitutes the main source of protease activities within CPs. Together, these results suggest that CPs generate plasmin, mainly in the core, which could in turn participate in MMP activation and the onset of complications. PMID- 16713601 TI - The haemoxisome: a haem-iron containing structure in the Rhodnius prolixus midgut cells. AB - Rhodnius prolixus midgut was analysed using transmission electron microscopy and electron spectroscopic imaging in order to localize the cellular structures involved in haem metabolism. In the posterior midgut, special cellular electron dense structures were observed. These structures are here designated haemoxisomes. Haemoxisomes are present in the epithelial cells at various time points after a blood meal. Several days after the blood meal, some of them become less electron-dense. By electron spectroscopic imaging, large amounts of iron and oxygen were detected in these cellular structures. The iron is probably bound to the porphyrin ring as an iron-protoporphyrin IX complex, as detected using the diaminobenzidine technique. An interesting observation was the presence of endoplasmic reticulum surrounding the haemoxisomes during some special periods. Iron content was monitored in the posterior midgut epithelium and was found to be constant at the initial days after a blood meal, but slightly higher at the end of the digestive process (from 13th up to 20th day). These results are in agreement with the observation that the appearance of the haemoxisomes changes at the end of the digestive process. The ability to degrade haem seems to depend on the presence of endoplasmic reticulum as observed using a haem degradation assay in the presence of an endoplasmic reticulum-enriched fraction. Taken together these results suggest that haemoxisomes may play a role in intracellular haem detoxification. PMID- 16713602 TI - A new Bombyx mori larval ovarian cell line highly susceptible to nucleopolyhedrovirus. AB - Lepidopteran cell lines constitute the backbone for studying baculoviral biology in culturo and for baculovirus vector based recombinant protein expression systems. In the present study, we report establishment of a new continuous cell line designated as DZNU-Bm-1 from larval ovaries of the silkworm, Bombyx mori. The cells were grown in MGM-448 insect cell culture medium supplemented with 10% fetal bovine serum (FBS) and 3% heat inactivated B. mori haemolymph at 25+/-1 degrees C. A large number of attached epithelial-like and round refractive cells migrated from the explants and multiplied in the primary cultures. Both type of cells were subcultured initially for a few passages but after 10 passages the round refractive cells dominated the population, which could be subcultured continuously using MGM-448 medium with 10% FBS. The population doubling time of cell line was about 42h at 25+/-1 degrees C. The cell populations were largely diploids and triploids, while a few tetraploids and hexaploids were also observed. DNA profiles using Inter Simple Sequence Repeat (ISSR)-PCR and Simple Sequence Repeat (SSR) loci established the differences between DZNU-Bm-1 cell line and most widely used BmN cell line and the B. mori W-chromosome specific sequences confirmed the origin of DZNU-Bm-1 cell line to be from female silkworm. When cells were infected with free nonoccluded B. mori nucleopolyhedrovirus (BmNPV), the cell line was found to be highly susceptible with 92-94% of the cells harbouring BmNPV and having an average of 20-23 OBs/infected cell. We suggest the usefulness of this cell line in BmNPV based baculoviral expression system and also for studying in culturo virus replication. PMID- 16713604 TI - Inhibition of cyclooxygenase-2 improves cardiac function following long-term preservation. AB - BACKGROUND: Cyclooxygenase (COX) is an intracellular enzyme that converts arachidonic acid to prostaglandin endoperoxide (PGG(2)). There are two isoforms of COX, namely constitutive COX-1 and inducible COX-2. It has been reported that COX-2 plays an important role in ischemia-reperfusion injury and that COX-2 mRNA and protein expression were up-regulated during cardiac allograft rejection. FK3311 is a suppressor of COX-2 activation. The purpose of this study was to evaluate the effectiveness of inhibiting COX-2 with FK3311 for the minimization of ischemia-reperfusion injury and for the improvement of donor heart function following transplantation in a canine model. MATERIALS AND METHODS: Adult mongrel dogs were used. After the measurement of hemodynamic parameters [cardiac output (CO), left ventricular pressure (LVP), and the maximum rates of increase and decrease in LVP (+/-LVdp/dt)], coronary vascular beds were washed out with a hypothermic (4 degrees C) University of Wisconsin (UW) solution following cardiac arrest in response to cold (4 degrees C) glucose-insulin-potassium solution. The heart was then excised and preserved in hypothermic (4 degrees C) UW solution for 12 h. FK3311 (3 mg/kg) was administered intravenously to five dogs prior to reperfusion, while vehicle was administered intravenously to a control group (n = 5). After 3 h of orthotopic transplantation using cardiopulmonary bypass, the hemodynamic parameters were compared with preoperative values of the donor animals under the condition of 10 mm Hg right atrial pressure and 5 mug/kg/min dopamine support. RESULTS: The recovery rates of CO and +/-LVdP/dt were significantly (P < 0.05) higher in the FK-treated dogs than in the controls (CO: 93 +/- 6 versus 66% +/- 4%; +LVdp/dt: 125 +/- 8 versus 77 +/- 10%; and -LVdp/dt: 81 +/- 7 versus 52 +/- 6%; for FK-treated versus control dogs, respectively). The recovery rate of LVP was higher in the FK-treated dogs than in the controls (90 +/- 5 versus 72 +/- 5%), but this difference was not statistically significant. Immunohistochemical staining revealed that COX-2 expression was reduced significantly in the myocardium of FK-treated dogs compared with controls. CONCLUSION: Hemodynamic parameters following transplantation were improved significantly in dogs treated with FK3311. Therefore, the inhibition of COX-2 improves transplanted cardiac function following long-term preservation. PMID- 16713603 TI - Salvianolic acid B from Salvia miltiorrhiza inhibits tumor necrosis factor-alpha (TNF-alpha)-induced MMP-2 upregulation in human aortic smooth muscle cells via suppression of NAD(P)H oxidase-derived reactive oxygen species. AB - Activated matrix metalloproteinases (MMPs) in patients with acute coronary syndromes may contribute to plaque destabilization. Tumor necrosis factor-alpha (TNF-alpha) enhances NAD (P) H oxidase-dependent reactive oxygen species (ROS) formation and ROS induce MMP-2. In the present study, the effects of a potent water-soluble antioxidant, salvianolic acid B (SalB), derived from a Chinese herb, Salvia miltiorrhiza, on the expression of MMP-2 by TNF-alpha-treated human aortic smooth muscle cells (HASMCs) were investigated. In this study, salvianolic acid B scavenged H2O2 in a dose-dependent manner in test tube. We found that SalB, as well as NADPH oxidase inhibitors, DPI or apocynin, and antioxidant NAC, inhibited TNF-alpha-induced MMP-2 mRNA, protein expression, and gelatinolytic activity in HASMCs in a concentration-dependent manner. We also observed a dose dependent decrease in ROS production and NADPH oxidase activity induced by TNF alpha in the presence of SalB. SalB also significantly inhibited angiotensin II or H2O2-induced MMP-2 mRNA and protein expression and gelatinolytic activity in HASMCs. Our data point out that the importance of NADPH oxidase-dependent ROS generation in the control of SalB inhibition of TNF-alpha-induced MMP-2 expression and activity. PMID- 16713605 TI - Association of prostate-specific membrane antigen with caveolin-1 and its caveolae-dependent internalization in microvascular endothelial cells: implications for targeting to tumor vasculature. AB - Prostate-specific membrane antigen (PSMA) is a transmembrane protein with a highly restricted profile of expression. Expression is primarily limited to secretory cells of the prostatic epithelium, with elevated levels observed in prostate cancer. As an integral membrane protein correlated with prostate cancer, PSMA offers a potentially valuable target for immunotherapy. PSMA is also detected in the neovasculature of a variety of solid tumors but not in the endothelial cells of preexisting blood vessels. Although the significance of PSMA expression in these cells remains elusive, this pattern of expression implies that PSMA may perform a functional role in angiogenesis and may offer a therapeutic target for the treatment of a broad spectrum of solid tumors. In this study, we have expressed PSMA in human microvascular endothelial cells and demonstrate that PSMA binds to caveolin-1 and undergoes internalization via a caveolae-dependent mechanism. The association between PSMA and caveolae in endothelial cells may provide important insight into PSMA function and ways to best exploit this protein for therapeutic benefit. PMID- 16713606 TI - Tolerance to morphine analgesia: evidence for stimulus intensity as a key factor and complete reversal by a glycine site-specific NMDA antagonist. AB - N-methyl-D-aspartate (NMDA) receptors are widely involved in opioid tolerance. However, it is less clear whether NMDA receptor antagonists reverse already established tolerance and whether the intensity of the nociceptive stimulus influences morphine tolerance. Three days after implantation of morphine or control pellets the effects of i.v. morphine and pre-administration of saline or (+)-HA966 (a glycine site-specific NMDA receptor antagonist), were studied on the C-fibre reflex elicited by a wide range of stimulus intensities. Morphine both increased the threshold and decreased the slope of the recruitment curve in the "non-tolerant" group of animals. In the "morphine-tolerant" group, the threshold did not change but the gain of the stimulus-response curve decreased. The expression of tolerance to morphine depended on the intensity of the stimulus, being maximal when threshold stimulus intensities were used but considerably less with supra-threshold stimulation. As expected, a single treatment with (+)-HA966, potentiated morphine antinociception in "non-tolerant" rats. However, in "morphine-tolerant" rats (+)-HA966 reversed established morphine tolerance and increased the antinociceptive effects of morphine. These results suggest that (+) HA966 interfered with expression of morphine tolerance, and offered an encouraging therapeutic approach for pain management in opioid abusers. PMID- 16713607 TI - Modulation of thalamic neuron excitability by orexins. AB - Orexins (hypocretins) are peptides of hypothalamic origin that play an important role in maintaining wakefulness. Reduced orexin levels have been associated with an increased incidence of narcolepsy. Considering thalamic nuclei are interconnected with virtually all neocortical regions and the thalamus has been found to produce distinct activities related to different levels of arousal, we have examined the actions of orexins on thalamic neurons using an in vitro thalamic slice preparation. The orexins (orexin-A and orexin-B) produced distinct actions within different intralaminar nuclei. Orexin-B strongly depolarized the majority of centrolateral nucleus (CL) neurons (71%), but depolarized a significantly smaller population of parafascicular nuclei (Pf) neurons (10%). In the mediodorsal thalamic nucleus (MD), orexin-B depolarized 21% of the neurons tested. Overall, orexin-B was found to be more potent than Orexin-A. Orexin-A depolarized a significantly smaller population of CL neurons (23%), but had no effect on Pf neurons. In addition, orexin-A produced a small depolarization in 28% of neurons in the thalamic reticular nucleus (TRN). Both orexin-A and orexin B had no effect on neurons in the lateral posterior (LP), lateralodorsal (LD), posterior thalamic (Po), ventrobasal (VB) nucleus and lateral geniculate nucleus (LGN). The depolarizing actions of orexins were sufficient to alter the firing mode of these neurons from a burst- to tonic-firing mode. The excitatory actions of orexin-B result from a decrease in the apparent leak potassium current (Kleak). The orexin-B mediated excitation was also attenuated by bupivacaine suggesting the involvement of Kleak current. Further, the actions of orexin-B were occluded by the classical neurotransmitter dopamine, indicating the orexins may share similar ionic mechanisms. Thus, the depolarizing actions of orexins may play a key role in altering the firing mode of thalamic neurons associated with different states of consciousness. PMID- 16713608 TI - Influenza-related deaths and hospitalizations in Hong Kong: a subtropical area. AB - BACKGROUND: While influenza vaccination has been widely used in developed countries to reduce mortality and morbidity in high-risk populations, the lack of regional data on the health burden of influenza and the uncertainty of the applicability of data from temperate areas have been major impediments to establishing an evidence-based policy on the wider use of influenza vaccine in tropical and subtropical regions of the world. OBJECTIVES: The aim of this study was to estimate the annual excess deaths and hospitalizations related to influenza in the Hong Kong Special Administrative Region (HKSAR) for the years 1999 and 2000. STUDY DESIGN AND METHODS: Correlation and regression models were used to estimate the excess deaths and hospitalizations related to influenza in the general population of HKSAR for the years 1999 and 2000, using routinely collected mortality and hospitalization data, and virological laboratory data collected by the HKSAR Influenza Surveillance System. RESULTS: The annual mean excess numbers of deaths related to influenza in Hong Kong were estimated to be 613 for pneumonia and influenza, and 2302 for respiratory and circulatory diseases. The mean excess numbers of hospitalizations attributable to influenza were 4051 for pneumonia and influenza, and 15,873 for respiratory and circulatory diseases. The crude influenza-related mortality and hospitalization rates in Hong Kong, a subtropical area, exceeded those documented in temperate regions. CONCLUSIONS: The finding of significant mortality and morbidity related to influenza in a subtropical area is in accordance with the results of previous studies in tropical and subtropical regions. This simple methodology can be used for the development of influenza immunization policy in many developing countries in tropical and subtropical regions. The enormous potential of influenza vaccination in saving lives and reducing suffering warrants serious consideration of the expanded use of influenza vaccine in tropical and subtropical regions. PMID- 16713609 TI - Frequency and gravity of human envenomations caused by marine catfish (suborder siluroidei): a clinical and epidemiological study. AB - INTRODUCTION AND OBJECTIVES: Catfish occur in marine and freshwater environments worldwide. They have three serrated venomous bony stings in the dorsal and pectoral fins that are used for defence against predators and are refilled by glandular tissues under the epithelium. However, some catfishes do not have poisonous glands next to the sting and cause traumatic wounds without poisoning. The objective of this study was to provide data for, and comment on, the epidemiological and clinical problems caused by marine catfish. PATIENTS AND METHODS: The authors have observed, followed and documented 127 injuries caused by marine catfish stings during different phases of the envenoming over a time period of 8 years at three points along the Western Atlantic Ocean coast. RESULTS: The patients presented intense pain during the acute phase of envenoming and complications, such as bacterial and fungi infections and retention of bony fragments, in the later phase. Immersion of the affected extremity in hot water was used in about 20% of cases with excellent results. DISCUSSION: Injuries caused by marine catfish are common (about 20% of injuries caused by marine animals in a series of more than 700 injuries recorded by the author) and cause intense pain and later complications. Immersion of the affected extremity in hot water results in improvement in the acute phase, but does not prevent the appearance of secondary infection or foreign body reactions. PMID- 16713610 TI - Induction of the SOS response by bacteriophage lytic development in Salmonella enterica. AB - Infection of Salmonella enterica with lytic mutants of either P22 or SE1 bacteriophages triggers the expression of its DNA damage-inducible SOS response through a lexA-dependent pathway. This induction of the SOS system strictly requires the presence of the bacteriophage kil gene. Accordingly, plasmid overexpression of the kil gene also promotes the S. enterica SOS network induction. Furthermore, S. enterica Gifsy prophages are induced following the infection with SE1 and P22 lytic derivatives. The observed data reveal a hitherto unknown SOS system-mediated fail-safe mechanism of resident prophages against infection with heteroimmune lytic bacteriophages and suggest a novel role for the kil family of proteins. PMID- 16713611 TI - In vitro selected Con1 subgenomic replicons resistant to 2'-C-methyl-cytidine or to R1479 show lack of cross resistance. AB - The HCV polymerase is an attractive target for the development of new and specific anti-HCV drugs. Herein, the characterization of the inhibitory effect of 2'-C-Methyl-Cytidine shows that it is a potent inhibitor of both genotype 1b and 1a HCV replicon replication, both of laboratory-optimized as well as of NS5B clinical isolates-chimera replicons. The corresponding 5'-triphosphate derivative is a potent inhibitor of native HCV replicase isolated from replicon cells and of the recombinant genotype 1b and 1a HCV polymerase-mediated RNA synthesis. Resistance to 2'-C-Methyl-Cytidine was mapped to amino acid substitution S282T in the NS5B coding region. Cross-resistance was observed to 2'-C-Methyl-Adenosine but not to interferon alpha-2a, to non-nucleoside HCV polymerase inhibitors or to R1479, a new and potent nucleoside inhibitor of NS5B polymerase. In vitro studies mapped resistance to R1479 to amino acid substitutions S96T and S96T/N142T of the NS5B polymerase. These mutations did not confer resistance to 2-C-Methyl Cytidine, thus confirming the lack of cross-resistance between these two HCV inhibitors. These data will allow the optimization of new polymerase inhibitors and their use in combination therapy. PMID- 16713613 TI - Development of an improved PCR-ICT hybrid assay for direct detection of Legionellae and Legionella pneumophila from cooling tower water specimens. AB - A novelly improved polymerase chian reaction and immunochromatography test (PCR ICT) hybrid assay comprising traditional multiplex-nested PCR and ICT, (a lateral flow device) was developed for direct detection of Legionella bacteria from environmental cooling tower samples. The partial 16S rDNA (specific for Legionella spp.) and dnaJ (specific for Legionella pneumophila) genes from Legionella chromosome were first specifically amplified by multiplex-nested PCR, respectively, followed by detection using ICT strip. Reading of results was based on presence or absence of the two test lines on the strips. Presence of test line 1 indicated existence of Legionella spp. specific 16S rDNA and identified Legionella spp. Presence of test line 2 further indicated existence of dnaJ and thus specifically identified L. pneumophila. In contrast, for non-Legionellae bacteria no test line formation was observed. Results of direct detection of Legionella bacteria and L. pneumophila from water tower specimens by this assay showed 100% sensitivity, and 96.6% and 100% specificity, respectively compared with traditional culture, biochemical and serological identification methods. The PCR-ICT hybrid assay does not require sophisticated equipment and was proved to be practically useful in rapid and direct Legionellae detection from environmental water samples. PMID- 16713614 TI - Distribution of polycyclic aromatic hydrocarbons in soils at Guiyu area of China, affected by recycling of electronic waste using primitive technologies. AB - The concentration, distribution, profile and possible source of polycyclic aromatic hydrocarbons (PAHs) in soil were studied in Guiyu, an electronic waste (E-waste) recycling center, using primitive technologies in Southeast China. Sixteen USEPA priority PAHs were analyzed in 49 soil samples (0-10 cm layer) in terms of individual and total concentrations, together with soil organic matter (SOM) concentrations. The concentrations of a sum of 16 PAHs ranged from 44.8 to 3206 microgkg(-1) (dry weight basis), in the descending order of E-waste open burning sites (2065 microgkg(-1))>areas near burning sites (851microgkg(-1))>rice fields (354 microgkg(-1))>reservoir areas (125microgkg(-1)). The dominant PAHs were naphthalene, phenanthrene and fluoranthene, which were mainly derived from incomplete combustion of E-waste (e.g. wire insulations and PVC materials), and partly from coal combustion and motorcycle exhausts. All individual and total PAH concentrations were significantly correlated with SOM except for naphthalene and acenaphthylene. Principal component analysis was performed, which indicated that PAHs were mainly distributed into three groups in accordance with their ring numbers and biological and anthropogenic source. In conclusion, PAH concentrations in the Guiyu soil were affected by the primitive E-waste recycling activities. PMID- 16713615 TI - Occurrence of polychlorinated biphenyls (PCBs) in the Soulou stream in the power generation area of Eordea, northwestern Greece. AB - The occurrence of polychlorinated biphenyls (PCBs) was investigated in the Soulou stream in the area of Eordea, northwestern Greece, receiving the effluents of two lignite burning power plants. The study was carried out after an accidental fire in one of the power plants during which losses of an amount of Aroclor 1232 were recorded. PCBs were determined in water and sediment samples collected from the wastewater treatment unit of the power plant and various points across the stream. Concentrations of PCBs were in the same order of magnitude as in other surface waters and sediments previously found in the Greek territory. The sum concentrations of seven PCBs (#28, #52, #101, #118, #138, #153, #180) varied between 94 and 206 ngl(-1) in waters, and between 67 and 500 ngg(-1) in sediments. Profile analysis of PCB congeners revealed large differences between waters and sediments, showing also significant dissimilarity with the profile of Aroclor 1232. PMID- 16713616 TI - Development of an analytical method for the determination of anthracyclines in hospital effluents. AB - Little is known about the fate of cytostatics after their elimination from humans into the environment. Being often very toxic compounds, their quantification in hospital effluents may be necessary to individualise the putative magnitude of pollution problems. We therefore developed a method for the determination of the very important group of anthracyclines (doxorubicin, epirubicin, and daunorubicin) in hospital effluents. Waste water samples were enriched by solid phase extraction (concentration factor 100), analysed by reversed-phase high performance liquid chromatography (RP-HPLC), and monitored by fluorescence detection. This method is reproducible and accurate within a range of 0.1-5 micro g l(-1) for all compounds (limits of quantification: 0.26-0.29 micro g l(-1) ; recoveries >80%). The applicability of the method was proven by chemical analysis of hospital sewage samples (range: 0.1-1.4 micro g l(-1) epirubicin and 0.1-0.5 micro g l(-1) doxorubicin). Obtained over a time period of one month, the results were in line with those calculated by an input-output model. These investigations show that the examined cytostatics are easily detectable and that the presented method is suitable to estimate the dimension of pharmaceutical contamination originating from hospital effluents. PMID- 16713612 TI - Evolution and adaptation of H5N1 influenza virus in avian and human hosts in Indonesia and Vietnam. AB - Highly pathogenic avian influenza virus H5N1 is endemic in poultry in East and Southeast Asia with disease outbreaks recently spreading to parts of central Asia, Europe and Africa. Continued interspecies transmission to humans has been reported in Vietnam, Thailand, Cambodia, Indonesia and China, causing pandemic concern. Here, we genetically characterize 82 H5N1 viruses isolated from poultry throughout Indonesia and Vietnam and 11 human isolates from southern Vietnam together with sequence data available in public databases to address questions relevant to virus introduction, endemicity and evolution. Phylogenetic analysis shows that all viruses from Indonesia form a distinct sublineage of H5N1 genotype Z viruses suggesting this outbreak likely originated from a single introduction that spread throughout the country during the past two years. Continued virus activities in Indonesia were attributed to transmission via poultry movement within the country rather than through repeated introductions by bird migration. Within Indonesia and Vietnam, H5N1 viruses have evolved over time into geographically distinct groups within each country. Molecular analysis of the H5N1 genotype Z genome shows that only the M2 and PB1-F2 genes were under positive selection, suggesting that these genes might be involved in adaptation of this virus to new hosts following interspecies transmission. At the amino acid level 12 residues were under positive selection in those genotype Z viruses, in the HA and PB1-F2 proteins. Some of these residues were more frequently observed in human isolates than in avian isolates and are related to viral antigenicity and receptor binding. Our study provides insight into the ongoing evolution of H5N1 influenza viruses that are transmitting in diverse avian species and at the interface between avian and human hosts. PMID- 16713618 TI - Hyaluronate and its receptors in bone marrow. AB - Cell-cell and cell-matrix interactions, which are mediated by cell adhesion molecules, play a fundamental role during many cellular processes including growth, differentiation, cell migration and cancer metastasis. One molecule playing a major role in these processes is the CD44 surface receptor, which is expressed in a wide range of cells including many cells of the hemopoietic system, where it mediates the interaction with its major ligand, hyaluronate. However, little is known about CD44 and hyaluronate in bone marrow and this was investigated immunohistochemically in trephine biopsies and in cultivated human bone marrow stromal cells. In biopsy specimens, patches of hyaluronate deposition were detected in the extracellular matrix (ECM). However, most of the areas of the ECM were devoid of hyaluronate. Single mast cells and lymphocytes scattered throughout the marrow were CD44 immunopositive. Marrow-derived stromal cells (MDSC) expanded in cell culture were immunopositive for CD44, hyaluronate synthase, and hyaluronate. Hence, a marked difference between CD44 immunolocalisation and hyaluronate deposition can be observed between in situ and under cell culture conditions. Since in normal marrow in situ the number of CD44 immunopositive cells was low, interactions of CD44 and hyaluronate would appear to not to play a major role in cell adhesion in the normal bone marrow. PMID- 16713619 TI - New technologies for the study of carnivore reproduction. AB - Routine analysis of urinary metabolites of estrogen and progesterone provided substantial information about the estrous cycle of bears. However, these data alone were not adequate to determine the precise timing of ovulation needed to maximize AI success rates, or to distinguish between pregnancy and pseudopregnancy. Therefore, there is a critical need to develop technologies that will enhance understanding of the reproductive mechanisms of ursids. Using the domestic dog as a model, three techniques were investigated for potential application to the propagation of captive endangered bears. In a modification of standard staining of bitch vaginal cells, trichrome staining of giant panda cells revealed two consistent chromic shifts 9 and 2 days prior to the periovulatory decrease in urinary estrone sulfate, enhancing the ability to predict ovarian events preceding ovulation. To further define the relationship between the decrease in estrogen and ovulation, the utility of a rapid immunochromatographic LH assay was investigated for giant pandas using a commercial LH kit canine serum. Serum collected during estrus exhibited positive test results, indicating the cross-reactivity of giant panda LH with canine LH antibodies, and preliminary data supported further development of the LH kit for the detection of LH in bear urine. Due to the limitations of hormone analysis for distinguishing pregnancy from pseudopregnancy in canids and ursids, forward-looking infrared thermography was evaluated as a method to visualize proliferating placental tissue, fetuses, or both. While further investigation is needed to confirm the utility of thermal imaging for pregnancy diagnosis in the domestic bitch, pregnancy and pseudopregnancy were successfully detected in two giant pandas. PMID- 16713617 TI - Stress resistance and aging: influence of genes and nutrition. AB - Previous studies have shown that dermal fibroblast cell lines derived from young adult mice of the long-lived Snell dwarf (dw/dw), Ames dwarf (df/df) and growth hormone receptor knockout (GHR-KO) mouse stocks are resistant, in vitro, to the cytotoxic effects of hydrogen peroxide, cadmium, ultraviolet light, paraquat, and heat. Here we show that, in contrast, fibroblasts from mice on low-calorie (CR) or low methionine (Meth-R) diets are not stress resistant in culture, despite the longevity induced by both dietary regimes. A second approach, involving induction of liver cell death in live animals using acetaminophen (APAP), documented hepatotoxin resistance in the CR and Meth-R mice, but dw/dw and GHR-KO mutant mice were not resistant to this agent, and were in fact more susceptible than littermate controls to the toxic effects of APAP. These data thus suggest that while resistance to stress is a common characteristic of experimental life span extension in mice, the cell types showing resistance may differ among the various models of delayed or decelerated aging. PMID- 16713620 TI - Predicting change over time in career planning and career exploration for high school students. AB - This study assessed 166 high school students in Grade 8 and again in Grade 10. Four models were tested: (a) whether the T1 predictor variables (career knowledge, indecision, decision-making self efficacy, self-esteem, demographics) predicted the outcome variable (career planning/exploration) at T1; (b) whether the T1 predictor variables predicted the outcome variable at T2; (c) whether the T1 predictor variables predicted change in the outcome variable from T1-T2; and (d) whether changes in the predictor variables from T1-T2 predicted change in the outcome variable from T1-T2. Strong associations (R(2)=34%) were identified for the T1 analysis (confidence, ability and paid work experience were positively associated with career planning/exploration). T1 variables were less useful predictors of career planning/exploration at T2 (R(2)=9%; having more confidence at T1 was associated with more career planning/exploration at T2) and change in career planning/exploration from T1-T2 (R(2)=11%; less confidence and no work experience were associated with change in career planning/exploration from T1 T2). When testing effect of changes in predictor variables predicting changes in outcome variable (R(2)=22%), three important predictors, indecision, work experience and confidence, were identified. Overall, results indicated important roles for self-efficacy and early work experiences in current and future career planning/exploration of high school students. PMID- 16713621 TI - Oil well produced water discharges to the North Sea. Part I: comparison of deployed mussels (Mytilus edulis), semi-permeable membrane devices, and the DREAM model predictions to estimate the dispersion of polycyclic aromatic hydrocarbons. AB - The oil companies operating in the Norwegian sector of the North Sea have conducted field studies since the mid-1990s to monitor produced water discharges to the ocean. These studies have been used to refine monitoring methods, and to develop and validate a dispersion and impact assessment model. This paper summarizes monitoring data from surveys conducted in two major oil and gas production areas, and compares the results to concentrations of polycyclic aromatic hydrocarbons (PAH) in surface waters predicted by the dose-related risk and effect assessment model (DREAM). Blue mussels and semi-permeable membrane devices (SPMDs) were deployed in the Ekofisk and Tampen Regions and analyzed for more than 50 PAH. PAH concentrations in ambient seawater were estimated based on the mussels and SPMD concentrations, and compared to model predictions. Surface water total PAH concentrations ranged from 25 to 350 ng/L within 1 km of the platform discharges and reached background levels of 4-8 ng/L within 5-10 km of the discharge; a 100,000-fold dilution of the PAH in the discharge water. The PAH concentrations in surface water, predicted by three methods, compared well for the Ekofisk Region. The model predicted higher concentrations than the field based methods for parts of the Tampen Region; particularly the most tidally influenced areas. Tidally-mediated fluctuations in PAH concentrations in surface water must be considered because they affect the estimation of PAH concentrations from mussel and SPMD residue data, and the predictions by the DREAM model. Predictions using mussels, SPMDs, and modeling support and complement each other; all are valuable tools for estimating the fate and impact of chemical contaminants in produced water that are discharged to the ocean. PMID- 16713622 TI - Effects of cadmium, zinc and nitrogen status on non-protein thiols in the macroalgae Enteromorpha spp. from the Scheldt Estuary (SW Netherlands, Belgium) and Thermaikos Gulf (N Aegean Sea, Greece). AB - Enteromorpha prolifera (Scheldt Estuary) and E. linza (Thermaikos Gulf) were incubated at three salinities with 100 and 200microgL(-1)Cd and Zn. The objective was to measure effects of Cd, Zn and nitrogen (N) status on the pools of metal binding non-protein thiols: glutathione and phytochelatins, (gamma-glutamyl cysteinyl)(n)-glycine (PC). In E. linza, ammonium pools were higher, but amino acid pools, total N and protein contents were lower than in E. prolifera. Reduced glutathione (GSH) pools were positively correlated with free glutamate and protein contents. In E. linza GSH pools increased and the ratio of reduced to oxidized glutathione (GSH:(GSH+0.5GSSG)), an indicator of oxidative stress, decreased with Cd contents, indicating Cd-induced glutathione oxidation. Total glutathione pools (reduced plus oxidized) ranged from 16nmolSgdwt(-1) in controls (at 0.5micromolCdgdwt(-1)) to 179nmolSgdwt(-1) (at 1.9micromolCdgdwt(-1)) at the highest cadmium dosage. Cadmium stimulated PC synthesis in E. prolifera which suggests that in N-rich algae, glutathione pools were high enough for PC synthesis. In both species GSH and protein increased with Zn contents, whereas GSH:(GSH+0.5GSSG) decreased, which would indicate Zn-induced oxidative stress; in E. linza, at the highest salinity the glutathione redox ratio decreased from 0.61 (at 2.9micromolZngdwt(-1)) to 0.26 (at 4.9nmolSgdwt(-1)) (at 0.5molCdgdwt(-1)). PCs were not synthesized in response to Zn, which may have resulted in Zn-induced GSH oxidation. The presence of both oxidative effects (Cd, Zn) and detoxification (Cd) could be identified by observing the responses of glutathione and PC pools to metal stress. PMID- 16713623 TI - Biomimetic synthesis of calcium-deficient hydroxyapatite in a natural hydrogel. AB - A novel composite material consisting of calcium-deficient hydroxyapatite (CdHAP) biomimetically deposited in a bacterial cellulose hydrogel was synthesized and characterized. Cellulose produced by Gluconacetobacter hansenii was purified and sequentially incubated in solutions of calcium chloride followed by sodium phosphate dibasic. A substantial amount of apatite (50-90% of total dry weight) was homogeneously incorporated throughout the hydrogel after this treatment. X ray diffractometry (XRD) showed that CdHAP crystallites had formed in the cellulose. XRD further demonstrated that the CdHAP was comprised of 10-50 nm anisotropic crystallites elongated in the c-axis, similar to natural bone apatite. Fourier transform infrared (FTIR) spectroscopy demonstrated that hydroxyl IR bands of the cellulose shifted to lower wave numbers indicating that a coordinate bond had possibly formed between the CdHAP and the cellulose hydroxyl groups. FTIR also suggested that the CdHAP had formed from an octacalcium phosphate precursor similar to physiological bone. Scanning electron microscopy (SEM) images confirmed that uniform approximately 1 microm spherical CdHAP particles comprised of nanosized crystallites with a lamellar morphology had formed in the cellulose. The synthesis of the composite mimics the natural biomineralization of bone indicating that bacterial cellulose can be used as a template for biomimetic apatite formation. This composite may have potential use as an orthopedic biomaterial. PMID- 16713624 TI - Dendrimer crosslinked collagen as a corneal tissue engineering scaffold: mechanical properties and corneal epithelial cell interactions. AB - Generation 2 polypropyleneimine octaamine dendrimers were used to generate highly crosslinked collagen with mechanical properties that would make it appropriate for use as a corneal tissue-engineering scaffold. Crosslinking of a highly concentrated collagen solution (2-4%) was effected using the water-soluble carbodiimide 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide hydrochloride (EDC). The multifunctional dendrimers were introduced as novel multifunctional crosslinkers after the activation of the carboxylic acid groups of glutamic and aspartic acid residues in collagen. Glutaraldehyde, a common collagen crosslinker, was used as comparison, as was EDC, itself an alternative crosslinker, which forms "zero-length or self-crosslinking". The mechanical properties resultant gels were determined. Young's modulus of the dendrimer crosslinked gels was significantly higher than that observed with the other crosslinkers, increasing to 5 MPa compared with 0.1 MPa for the EDC crosslinked gels. Transmission electron microscopy (TEM) analysis of the gels demonstrated the presence of fibrils in the thermally gelled collagen controls; no fibrils were observed in the dendrimer crosslinked gels. As a result, the optical transparency of the dendrimer crosslinked collagen was significantly better than that of the collagen thermal gels. The EDC and glutaraldehyde crosslinked gels were generally less transparent than those crosslinked with the dendrimers. Glucose permeation results demonstrated that the dendrimer crosslinked collagen had higher glucose permeability than natural human cornea. Dendrimer crosslinked collagen gels supported human corneal epithelial cell growth and adhesion, with no cell toxicity. In comparison, some potentially cytotoxic effects were observed with glutaraldehyde crosslinked collagen. Overall, the dendrimer crosslinked collagen gels showed promising properties that suggest that these might be suitable scaffolds for corneal tissue engineering and potentially other tissue engineering applications. PMID- 16713626 TI - Posterior corneal curvature changes after undersurface ablation of the flap and in-the-bed LASIK retreatment. AB - OBJECTIVE: To analyze LASIK retreatment-induced changes in the posterior corneal curvature (PCC) with undersurface ablation of the flap (UAF) and in-the-bed techniques. DESIGN: Nonrandomized, comparative, interventional study. PARTICIPANTS: Forty-six eyes with a residual spherical equivalent refraction between -0.37 and -2.75 diopters (D) and astigmatism between 0.0 and -1.25 D were included prospectively. In 23 eyes, the calculated postenhancement flap thickness exceeded 150 microm using micropachymetric optical coherence tomography, whereas with further ablation of the bed, the residual bed thickness (RBT) would have been <250 microm, or <55% of the pre-LASIK central pachymetry. In another 23 eyes, RBT allowed the planned ablation for a calculated post-retreatment RBT exceeding 250 microm, >55% of the pre-LASIK central pachymetry. INTERVENTION: Eyes with insufficient RBT for further ablation underwent UAF retreatment, whereas those with adequate RBT received conventional in-the-bed LASIK retreatment. Examinations were performed before retreatment and 3 and 6 months postoperatively. No eye was lost to follow-up. MAIN OUTCOME MEASURES: Micropachymetry, Orbscan II scanning-slit PCC data, and visual acuity (VA). RESULTS: The groups did not differ in age, intraocular pressure, or retreatment ablation depth, but the UAF eyes had a lower mean pre-retreatment RBT (270.7+/ 25.4 microm) than conventional enhancement eyes (353.0+/-41.5 microm) (P = 0.001). Eyes undergoing UAF had no significant change in PCC, whereas eyes undergoing conventional retreatment had an increase in the posterior corneal power within the central 3-mm zone (P = 0.008) 3 months after retreatment. No significant changes occurred thereafter. The amount of change in posterior corneal power within the 3-mm central zone from before to after retreatment differed significantly between the groups (mean difference, 0.135 D; 95% confidence interval, 0.022-0.248 D; P = 0.02). No keratectasia developed clinically, and no retreated eye lost or gained > or =2 lines of best-corrected VA. Six months after retreatment, the efficacy and safety indices for the UAF procedure were 0.96 and 1.01, respectively, and 1 and 1.06 for conventional LASIK enhancement. CONCLUSION: Undersurface ablation of the flap retreatment appears to have less potential for changing the posterior corneal surface than conventional LASIK enhancement and can help reduce the likelihood of retreatment-induced keratectasia. PMID- 16713627 TI - The influence of incision-induced astigmatism and axial lens position on the correction of myopic astigmatism with the Artisan toric phakic intraocular lens. AB - PURPOSE: To evaluate postoperative astigmatism with regard to incision-induced astigmatism and deviation in axial alignment with the use of preoperative limbal marking with the Javal keratometer (Haag Streit, Bern, Switzerland) in eyes implanted with the Artisan toric phakic intraocular lens (IOL) (Ophtec, Groningen, The Netherlands). DESIGN: Prospective nonrandomized trial. PARTICIPANTS: Fifty-four eyes of 33 patients with myopia (mean, -9.67 diopters [D]) and astigmatism (mean, -3.44 D). INTERVENTION: The enclavation site was marked on the limbus using the Javal keratometer. The Artisan toric phakic IOL was implanted according to the axis marked on the limbus. Follow-up was a minimum of 6 months. MAIN OUTCOME MEASURES: Safety index, efficacy index, predictability, safety, and vector analysis of total refractive correction were determined. The effects of axis misalignment and incision-induced astigmatism on the final refractive error were evaluated. RESULTS: At 6 months after surgery, the safety index was 1.29+/-0.29 and the efficacy index was 1.04+/-0.35. Mean spherical equivalent subjective refraction reduced from -11.39+/-4.86 D before surgery to 0.38+/-0.57 D at 6 months. Sixty-seven percent of eyes were within 0.50 D of attempted refraction and 89% were within 1.00 D. Mean preoperative cylinder was 2.92+/-1.60 D at 91.4 degrees . At 6 months, the mean cylinder was 0.28+/-0.54 D at 174.3 degrees . No eyes lost 2 or more lines of best-corrected visual acuity at 6 months. Eighty-three percent of eyes achieved uncorrected visual acuity of 20/40 and 28% achieved 20/20. Vector analysis of total surgically induced astigmatism revealed a mean cylindrical change of 3.21+/-1.71 D. Average axis misalignment was 0.37+/-5.34 degrees . The mean incision-induced astigmatism was 0.74+/-0.61 D at 0.2 degrees . CONCLUSIONS: Implantation of the myopic toric IOL leads to safe, efficacious, and predictable results. The level of unpredictability caused by minor axis IOL misalignment has minimal effects on the residual refractive error. The procedure of axis alignment with the Javal keratometer seems to be an accurate method of marking the eye for toric IOL implantation. Incision-induced astigmatism can result in an overcorrection of the cylinder. A systematic undercorrection of -0.50 D for attempted cylindrical outcome could result in an achieved correction closer to emmetropia. PMID- 16713628 TI - Intravitreal voriconazole for the treatment of endogenous Aspergillus endophthalmitis. AB - OBJECTIVE: To describe the first use of intravitreal voriconazole in a human eye for the treatment of Aspergillus endophthalmitis. DESIGN: Interventional case report. PATIENT: A 22-year-old woman receiving immunosuppressive agents 5 weeks after lung transplantation who presented with blurred vision and redness in the right eye. INTERVENTIONS: Intravitreal injection of voriconazole (100 microg/0.1 ml) with pars plana vitrectomy, given after isolation of A. terreus in the vitreous sample. Previous treatment modalities, including vitrectomy with repeated intravitreal amphotericin B and systemic voriconazole, failed to prevent deterioration. MAIN OUTCOME MEASURES: Visual acuity (VA) and ocular inflammation. RESULTS: Significant improvement was observed in VA (to 6/15) and in ocular inflammatory reaction. The patient recovered with no evidence of systemic fungal infection. CONCLUSION: Intravitreal voriconazole may be used as an adjunct to systemic treatment in patients with Aspergillus endophthalmitis. Further clinical studies are needed to determine how often this approach can safely treat this condition. PMID- 16713630 TI - Exposure to early life trauma is associated with adult obesity. AB - Exposure to traumatic events during childhood is associated with an elevated risk of adult obesity. It has been hypothesized that the psychological sequelae from childhood trauma account for this risk, though no study has examined whether an increased risk of obesity is found in persons without psychological disorders. We examined exposure to early life stressors and body mass index (BMI) in 696 adults without significant medical or psychiatric history. Bivariate correlation showed that the total number of early life stressors (r=0.08), age (r=0.19), and sex (r=0.16) were significantly related to adult BMI. Given the relationship between sex and BMI, we examined the contribution of early life stressors to adult obesity separately for men and women. In men, hierarchical regression showed that exposure to early life stressors predicted adult obesity. Specifically, history of being bullied/rejected (Obese 31%, Normal weight, 9%) and emotional abuse (Obese, 17%; Normal weight, 2%) predicted adult obesity after controlling for the effects of age. In women, no relationship between early life stressors and adult obesity was found. These findings suggest that multiple processes mediate the relationship between early life stress and adult obesity and that their relative contributions may differ between men and women. PMID- 16713629 TI - Antecedent life events of binge-eating disorder. AB - The present study investigated the occurrence of life events preceding the onset of disturbed eating in binge-eating disorder (BED). In a case-control design, 162 matched pairs of black and white women with BED and women with no current psychiatric disorder, and 107 matched pairs of women with BED and a current general psychiatric disorder were recruited from the community for the New England Women's Health Project. Life events in the year before the onset of disturbed eating were assessed retrospectively with an investigator-based interview. Women with BED reported exposure to a significantly greater number of life events during the year before onset of eating disturbances than both the non psychiatric and psychiatric control women during the same period of time in their lives. Women with BED had a significantly higher risk of exposure to certain specific life events (e.g., critical comments about shape, weight, or eating; stress related to work, school or other sources; major changes in life circumstances and relationships; physical abuse; and feeling unsafe in a variety of settings) than the non-psychiatric control women, while differences between the BED and the psychiatric control group were less marked. There was no evidence for race-specific exposure to antecedent life events. The results suggest that a greater number and certain specific types of life events increase risk for the subsequent development of BED. PMID- 16713631 TI - Determination of the carrier frequency of the common GJB2 (connexin-26) 35delG mutation in the Greek Cypriot population. AB - OBJECTIVE: Mutations in the GJB2 (connexin-26) gene are responsible for more than half of all cases of prelingual recessive inherited non-syndromic deafness in Europe. One specific mutation 35delG, accounts for up to 70% of the mutations detected in European populations and is one of the most frequent disease mutations identified so far. The aim of this study is to determine the percentage of carriers of this mutation in the Greek Cypriot population. METHODS: Genomic DNA was isolated from a total of 405 healthy unrelated Greek Cypriot adults. Screening for the frameshift 35delG mutation was performed by using an allele specific PCR protocol. Moreover, using the Poisson probability distribution, we compared the carrier frequencies of the 35delG mutation of the Greek Cypriot population to the various European and Middle Eastern populations. RESULTS: The carrier frequency in the Greek Cypriot population was estimated to be 2.5% and is similar to that observed in other European populations. The variance estimate for 35delG mutation produces slightly wider intervals with the Poisson model when compared with Binomial probability variance estimate. PMID- 16713632 TI - Development of real-time PCRs for detection and quantitation of human MMTV-like (HML) sequences HML expression in human tissues. AB - The human genome contains around 1000 betaretrovirus-like copies, human mouse mammary tumour virus (MMTV)-like (HML) groups 1-10, also referred to as human endogenous retrovirus "HERV-K". Despite many efforts, it is not established whether betaretroviruses, exo- or endogenous, are involved in the etiology of breast cancer, or other cancer diseases, in humans. Quantitative real-time PCR (QPCR) TaqMan-based assays for HML groups 1-7, targeting the conserved reverse transcriptase (RT) and integrase (IN) domains of the pol gene were designed. Plasmids containing the entire pol gene of HML1-7 were used as standards. The RT and IN based QPCRs could detect 10(0)-10(3) copies per PCR reaction of the plasmids. However, not all plasmids gave a signal in both RT and IN QPCRs, probably due to mismatches. Furthermore, RT and IN based HML6 specific QPCRs were developed. They were specific for amplification of transcripts for the whole HML6 group. The methods allow the monitoring in body fluids and tissues of expression of a wide range of betaretrovirus-like sequences. Betaretrovirus-like RNA was studied in normal human tissues and of HML6 in brains of multiple sclerosis (MS) patients. Brain, adrenal gland and testis had a high betaretrovirus-like expression. Multiple sclerosis plaques contained the same HML6 RNA concentration as control tissue. These assays are expected to enhance studies on involvement of betaretroviruses in physiology and disease. PMID- 16713633 TI - RT-PCR for detection of all seven genotypes of Lyssavirus genus. AB - The Lyssavirus genus includes seven species or genotypes named 1-7. Rabies genotypes correlate with geographical distribution and specific hosts. Co circulation of different lyssaviruses, imported cases, and the presence of unknown viruses, such as Aravan, Khujand, Irkut and West Caucasian Bat Virus, make it necessary to use generic methods able to detect all lyssaviruses. Primer sequences were chosen from conserved regions in all genotypes in order to optimise a generic RT-PCR. Serial dilutions of 12 RNA extracts from all seven Lyssavirus genotypes were examined to compare the sensitivity of the RT-PCR standardised in this study with a published RT-PCR optimised for EBLV1 detection and capable of amplifying RNA from all seven lyssaviruses. All seven genotypes were detected by both RT-PCRs, however, the sensitivity was higher with the new version of the test. Twenty samples submitted for rabies diagnosis were tested by the new RT-PCR. Eight out of 20 samples from six dogs, one horse and one bat were found positive, in agreement with immunofluorescence results. Seven samples from terrestrial mammals were genotype 1 and one from a bat was genotype 5. In conclusion, this method can be used to complement immunofluorescence for the diagnosis of rabies, enabling the detection of unexpected lyssaviruses during rabies surveillance. PMID- 16713634 TI - Early patterns of electrical activity in the developing cerebral cortex of humans and rodents. AB - During prenatal and early postnatal development, the cerebral cortex exhibits synchronized oscillatory network activity that is believed to be essential for the generation of neuronal cortical circuits. The nature and functional role of these early activity patterns are of central interest in neuroscience. Much of the research is performed in rodents and in vitro, but how closely do these model systems relate to the human fetal brain? In this review, we compare observations in humans with in vivo and in vitro rodent data, focusing on particular oscillatory activity patterns that share many common features: delta brushes, spindle bursts and spindle-like oscillations. There is considerable evidence that the basic functional properties of immature cortical networks are conserved through mammalian evolution, making the neonatal rodent an excellent model for studying early cortical activity and associated plasticity during the developmental period corresponding to the human fetal stage. This review is part of the INMED/TINS special issue "Nature and nurture in brain development and neurological disorders", based on presentations at the annual INMED/TINS symposium (http://inmednet.com/). PMID- 16713635 TI - Always returning: feedback and sensory processing in visual cortex and thalamus. AB - Feedback projections are an integral part of the mammalian visual system. Although it is tempting to relegate them to a subsidiary role in visual processing, because their supposed latency and lag might appear to be unfavourable for an involvement in fast processing, this is a dangerous simplification. Certainly for the world in motion, feedback from higher motion areas can influence the transfer of ascending input when, or even before, the input arrives. Here, we consider the circuit formed by layer 6 feedback cells in the visual cortex and how this straddles the retinothalamic and thalamocortical transfer of visual input. We discuss its links to feedback from the cortical motion area MT (V5), and suggest that motion perception involves a dynamic interplay between MT, V1 and the thalamus. This review is part of the TINS special issue on The Neural Substrates of Cognition. PMID- 16713636 TI - Building new motor responses: eyelid conditioning revisited. AB - Neural processes underlying memory and learning should be studied under the best possible physiological conditions - namely, in alert behaving animals. The classical conditioning of the nictitating membrane and eyelid response is a widely used experimental model for studying the neural bases of motor learning in mammals. Nevertheless, information is still needed on the functional aspects, taking place simultaneously in different cerebral structures, that underlie acquisition, extinction and recall of new motor and cognitive abilities. Here, we review recent data on the neural activity generated in selected brain sites (facial motor nuclei, deep cerebellar nuclei and the hippocampus) in simultaneity with the process of learning. The use of modern technologies for the proper recording of eyelid movements, for the identification of the recorded units, and for the activation of selective synaptic processes during the learning situation enables a precise redefinition of the role played by these neural structures in such associative learning. This review is part of the TINS special issue on The Neural Substrates of Cognition. PMID- 16713637 TI - The multipolar stage and disruptions in neuronal migration. AB - The genetic basis is now known for several disorders of neuronal migration in the developing cerebral cortex. Identification of the cellular processes mediated by the implicated genes is revealing crucial stages of neuronal migration and has the potential to reveal common cellular causes of neuronal migration disorders. We hypothesize that a newly recognized morphological stage of neuronal migration, the multipolar stage, is vulnerable and is disrupted in several disorders of neocortical development. The multipolar stage occurs as bipolar progenitor cells become radially migrating neurons. Several studies using in utero electroporation and RNAi have revealed that transition out of the multipolar stage depends on the function of filamin A, LIS1 and DCX. Mutations in the genes encoding these proteins in humans cause distinct neuronal migration disorders, including periventricular nodular heterotopia, subcortical band heterotopia and lissencephaly. The multipolar stage therefore seems to be a critical point of migration control and a vulnerable target for disruption of neocortical development. This review is part of the INMED/TINS special issue "Nature and nurture in brain development and neurological disorders", based on presentations at the annual INMED/TINS symposium (http://inmednet.com/). PMID- 16713638 TI - Local fluorouracil chemotherapy interferes with neural and behavioral recovery after brain tumor-like mass compression. AB - In this study, we investigated the impact of intracerebral delivery of chemotherapy on functional recovery from focal cortical tissue displacement, characteristic of brain tumors. Unilateral focal brain compression was induced by epidural implantation of an inverted hemisphere-shaped bead over the sensorimotor cortex. Microinjections of a total of 1mg chemoagent fluorouracil or the same volume of saline were made into the compressed cortex. Behavioral tests of forelimb sensorimotor function were conducted during 4 weeks' observation. Rats subjected to any of the three types of lesions, saline microinjection plus cortical compression, chemoagent microinjection alone, or chemoagent microinjection combined with cortical compression, demonstrated significant behavioral deficits in several sensorimotor tasks, compared with saline microinjected control animals. In placing tests, behavioral deficits elicited by each single treatment were worsened by combined treatment with chemoagent microinjection and focal cortical compression. Concurrently, local delivery of chemoagent into the compressed cortex induced increased cortical tissue loss, necrosis and apoptosis. These data indicate that local chemotherapy exacerbates compression-induced neurological impairment, and a model of controlled focal cortical compression may provide a valuable means to improve anti-cancer therapeutic designs with reduced deterioration of brain function. PMID- 16713639 TI - Genetic and dopaminergic modulation of reversal learning in a touchscreen-based operant procedure for mice. AB - Mice are uniquely suited as experimental subjects for various approaches to the study of the molecular and genetic basis of behavior, and there has been a corresponding explosion in the use of mice in behavioral neuroscience. Rats and monkeys, however, remain the preferred species for high-order cognitive models largely due to the unavailability of valid, reliable and translatable endpoint measures of behavior in the mouse. Here we present further development and validation of a touchscreen-based operant method for measuring cognition that is comparable to methods used in other species and human patients. C57BL/6J mice were found to show good performance on visual discrimination and reversal learning using this method. Demonstrating the sensitivity of the paradigm to genetic factors, C57BL/6J and DBA/2J mice exhibited marked differences in discrimination and reversal learning. Systemic treatment with the selective D1 like agonist, SKF81297, produced an impairment in the early phase of reversal learning, but did not alter visual discrimination, in C57BL/6J mice. The same treatment impaired spatial working memory on the T-maze delayed alternation task, but did not alter control measures of behavior including motivation and locomotor activity. These data demonstrate the sensitivity of visual discrimination and reversal learning measured by this method to genetic factors and pharmacological challenge, and thereby provide an extension and further validation of the method for measuring cognition in mice. When combined with emerging molecular techniques uniquely suited to this species such as genetic engineering and RNA modification this paradigm could provide a powerful new tool for behavioral neuroscience. PMID- 16713640 TI - Persistent neurochemical and behavioral abnormalities in adulthood despite early iron supplementation for perinatal iron deficiency anemia in rats. AB - BACKGROUND: Iron deficiency anemia (IDA) has been associated with altered cognitive, motor, and social-emotional outcomes in human infants. We recently reported that rats with chronic perinatal IDA, had altered regional brain iron, monoamines, and sensorimotor skill emergence during early development. OBJECTIVE: To examine the long-term consequences of chronic perinatal IDA on behavior, brain iron and monoamine systems after dietary iron treatment in rats. METHODS: Sixty dams were randomly assigned to iron-sufficient (CN) or low-iron (EID) diets during gestation and lactation. Thereafter, all offspring were fed the iron sufficient diet, assessed for hematology and behavior after weaning and into adulthood and for brain measures as adults (regional brain iron, monoamines, dopamine and serotonin transporters, and dopamine receptor). Behavioral assessments included sensorimotor function, general activity, response to novelty, spatial alternation, and spatial water maze performance. RESULTS: Hematology and growth were similar for EID and CN rats by postnatal day 35. In adulthood, EID thalamic iron content was lower. Monoamines, dopamine transporter, and dopamine receptor concentrations did not differ from CN. EID serotonin transporter concentration was reduced in striatum and related regions. EID rats had persisting sensorimotor deficits (delayed vibrissae-evoked forelimb placing, longer sticker removal time, and more imperfect grooming chains), were more hesitant in novel settings, and had poorer spatial water maze performance than CN. General activity and spatial alternation were similar for EID and CN. CONCLUSION: Rats that had chronic perinatal IDA showed behavioral impairments that suggest persistent striatal dopamine and hippocampal dysfunction despite normalization of hematology, growth and most brain measures. PMID- 16713641 TI - Up-regulation of the alligator CYP3A77 gene by toxaphene and dexamethasone and its short term effect on plasma testosterone concentrations. AB - In this study we describe an alligator hepatic CYP3A gene, CYP3A77, which is inducible by dexamethasone and toxaphene. CYP3A plays a broad role in biotransforming both exogenous compounds and endogenous hormones such as testosterone and estradiol. Alligators collected from sites in Florida that are contaminated with organochlorine compounds exhibit differences in sex steroid concentrations. Many organochlorine compounds induce CYP3A expression in other vertebrates; hence, CYP3A induction by organochlorine contaminants could increase biotransformation and clearance of sex steroids by CYP3A and provide a plausible mechanism for the lowering of endogenous sex steroid concentrations in alligator plasma. We used real time PCR to examine whether known and suspected CYP3A inducers (dexamethasone, metyrapone, rifampicin, and toxaphene) up-regulate steady state levels of hepatic CYP3A77 transcript to determine if induction patterns in female juvenile alligators are similar to those reported in other vertebrates and whether toxaphene, an organochlorine compound found in high concentrations in Lake Apopka alligators, induces this gene. Estrogen receptor alpha (ERalpha), estrogen receptor beta (ERbeta), androgen receptor (AR), glucocorticoid receptor (GR), progesterone receptor (PR), and steroid-xenobiotic receptor (SXR) transcripts were also measured to determine whether any of these nuclear receptors are also regulated by these compounds in alligators. Dexamethasone (4.2-fold) and toxaphene (3.5-fold) significantly induced CYP3A77 gene transcript, whereas rifampicin (2.8-fold) and metyrapone (2.1-fold) up regulated ERbeta after 24h. None of the compounds significantly up-regulated AR, ERalpha, GR, PR, or SXR over this time period. Plasma testosterone (T) did not change significantly after 24h in alligators from any of the treatment groups. Dexamethasone treated animals exhibited a strong relationship between the 24h plasma T concentrations and CYP3A77 (R(2)=0.9, positive) and SXR (R(2)=0.77, negative) transcripts, which suggests that the expression of these genes is related to plasma T in alligators. In light of our findings, we hypothesized that higher steady state CYP3A77 (and possibly SXR) gene expression would be observed in alligators collected from Lake Apopka, a polluted lake containing organochlorine compounds known to induce CYP3A isoforms in other taxa. Therefore, we measured basal levels of CYP3A77 and SXR gene transcripts in wild juvenile alligators collected from Orange Lake (reference lake), Lake Woodruff (reference lake), and Lake Apopka (contaminated lake). We found that no differences existed in CYP3A77 or SXR gene expression among animals from the lakes sampled suggesting that exposure to organochlorine compounds at concentrations present in Lake Apopka does not lead to variation in the expression of these genes, although capture stress could be interfering with these results since the glucocorticoid dexamethasone induces CYP3A77 transcript in alligators. PMID- 16713642 TI - The gene expression of adrenomedullin, calcitonin-receptor-like receptor and receptor activity modifying proteins (RAMPs) in CCl4-induced rat liver cirrhosis. AB - This study was undertaken to determine AM expression in carbon tetrachloride (CCl4)-induced liver cirrhosis developed with peritoneal ascites. Sprague-Dawley rats received subcutaneous injections of CCl4 twice weekly in olive oil (1:1, 0.3 ml per kg body weight) for 6 or 12 weeks until ascites developed, or saline in olive oil as control. At 6 weeks, fibrosis developed and at 12 weeks cirrhosis developed with ascites formation. At both 6 and 12 weeks, increases in plasma renin and AM were evident, as was the gene expression of AM. At 12 weeks after CCl4 injection, the gene expression of calcitonin-like-receptor (CRLR) and receptor activity modifying proteins (RAMP1, RAMP2 and RAMP3) were all elevated when compared to the control. The results suggest that liver cirrhosis increases mRNA expressions of AM, CRLR and RAMP1, RAMP2 and RAMP3 and that the increase in AM gene expression precedes the development of cirrhosis. The increase in AM synthesis as reflected by an increase in AM gene expression, together with a lack of increase in AM peptide at both 6 and 12 weeks may suggest an elevation of AM release. Given the potent vasodilatory action of AM, the increase in the synthesis and release of AM in the cirrhotic liver may also contribute to peripheral vasodilatation in liver cirrhosis. PMID- 16713643 TI - Intracerebroventricular injection of neuropeptide Y modifies carbohydrate and lipid metabolism in chicks. AB - The purpose of the present study was to investigate whether intracerebroventricular (ICV) injection of neuropeptide Y (NPY) affects heat production (HP), body temperature, and plasma concentrations of metabolic fuels in chicks. ICV injection of NPY (0, 188 or 375 pmol) did not affect HP, but significantly lowered respiratory quotient as well as the rectal temperature. These data suggest that the energy sources for HP were modified by NPY in the body. This idea was confirmed by subsequent experiments in which ICV injection of NPY significantly reduced plasma glucose and triacylglycerol concentrations but increased non-esterified fatty acid concentrations. The effect of NPY on the utilization of metabolic fuels was not associated changes in plasma catecholamine and corticosterone concentrations. In summary, the present study demonstrated that central NPY modifies peripheral carbohydrate and lipid metabolism in chicks. PMID- 16713644 TI - Interaction of bifidobacteria with Caco-2 cells-adhesion and impact on expression profiles. AB - The aim of the present study was to study different strains of bifidobacteria for adhesion to Caco-2 intestinal epithelial cells (IECs) and to test for the mRNA response of these cells following interaction with bifidobacteria. Adhesion was tested at different pH conditions using model epithelia consisting of transwell cultures of fully differentiated Caco-2 cells. Microarrays were used to characterize changes in global expression profiles of Caco-2 cells co-cultured with peripheral blood mononuclear cells (PBMCs) and challenged with non pathogenic Escherichia coli D2241 or four different strains of bifidobacteria. Furthermore, cytokine mRNA of IECs in responses to challenge with Bifidobacterium bifidum S17 or E. coli D2241 was tested in PBMC-sensitised Caco-2 cells using RT PCR. Bifidobacteria showed strain-specific adhesion to Caco-2. Shift of apical pH from 7 to 4.5 resulted in strain-specific changes of adhesion. Global expression profiles of PBMC-sensitised Caco-2 cells revealed differential expression of a significant number of genes only after challenge with E. coli D2241 while cells were essentially unresponsive to challenge with four strains of bifidobacteria showing different adhesion properties. Using a RT-PCR approach, in the same system a similar differential expression after challenge with E. coli D2241 or B. bifidum S17 was observed for various immune markers. The presented results suggest that Caco-2 cells might be specifically unresponsive to challenge with bifidobacteria irrespective of the level of adhesion. PMID- 16713645 TI - Black aspergilli and ochratoxin A in grapes in Italy. AB - The objective of this research was to investigate the presence of black aspergilli in grapes grown in Italy and to study the effect of environmental and cultural factors able to influence fungal incidence and ochratoxin A (OTA) presence. In this 3-year study, black aspergilli were frequently associated with grape berries; they were present in bunches starting from setting, colonising most berries at early veraison. Aspergillus carbonarius was never dominant at the different growth stages, or in different geographic areas and years, but it was confirmed as the key fungus because of the high percentage of strong OTA producer isolates in the population. The number of OTA producer strains, isolated in each vineyard at the different growth stages, was generally very limited and they were never statistically correlated to OTA content in bunches. The effect of geographic area on fungal flora was confirmed by statistical analysis, even though a major role was played by meteorological conditions, both on fungal colonisation and OTA content in bunches. Discriminant analysis gave promising perspectives for predicting OTA presence in vineyards in the future, based on summation of degree-day and rain in the period between 21st of August and 10th of September. PMID- 16713647 TI - Comparative genomics of Thermus thermophilus: Plasticity of the megaplasmid and its contribution to a thermophilic lifestyle. AB - The bacterium Thermus thermophilus grows at temperatures up to 85 degrees C and is equipped with thermostable enzymes of biotechnological interest. The recently decoded genomes of two strains of T. thermophilus, HB27 and HB8, each composed of a chromosome and a megaplasmid, must certainly encode specific strategies to encounter the thermophile challenge. Here, a genome comparison was undertaken to distinguish common functions from the flexible gene pool, which gave some clues about the biological traits involved in a thermophile lifestyle. The chromosomes were highly conserved, with about 100 strain-specific genes probably reflecting adaptations to the corresponding biological niche, such as metabolic specialities and distinct cell surface determinates including type IV pili. The two megaplasmids showed an elevated plasticity. Upon comparison and re-examination of their gene content, both megaplasmids seem to be implicated in assisting thermophilic growth: a large portion of their genes are apparently involved in DNA repair functions. About 30 plasmid-encoded genes exhibit sequence and domain composition similarity to a predicted DNA repair system specific for thermophilic Archaea and bacteria. Moreover, the plasmid-encoded carotenoid biosynthesis gene cluster is interlocked with genes involved in UV-induced DNA damage repair. This illustrates the importance of DNA protection and repair at elevated growth temperatures. PMID- 16713646 TI - Australian research on ochratoxigenic fungi and ochratoxin A. AB - The presence of the mycotoxin, ochratoxin A (OTA), has been reported in Australian grape products. Comprehensive surveys of Australian wines have determined that the frequency and level of OTA contamination are low. Aspergillus carbonarius is the primary OTA-producing species associated with grapes in Australia, and all isolates tested to date produce OTA. Aspergillus niger is isolated more frequently from vineyards, however, few strains produce OTA. A. carbonarius and A. niger exist as saprophytes in the top layer of soil beneath vines, from where they are thought to be blown onto bunches. The level of A. carbonarius in soil may be reduced by temperatures above or below the optimum temperature for survival (25 degrees C), by high soil moisture content, and by modifications to tillage and mulching practices. A. carbonarius is an opportunistic pathogen of damaged berries. In the absence of damage, spores may exist on berry surfaces without causing visible rots. Aspergillus rots are associated with black Aspergillus species, primarily A. niger, A. carbonarius and A. aculeatus. The potential for such rots is increased with berry damage, inoculum coverage and berry maturity. Susceptibility to berry splitting is related, in part, to bunch structure, and may be variety-dependent or influenced by rainfall, irrigation and canopy management. Black Aspergillus spp. are closely associated with berries near the main stem of the bunch. During winemaking, around 80% of the OTA initially present in grapes is removed, primarily with the skins and pulp during pressing. Additional reductions occur with the removal of precipitated grape and yeast solids. Bentonite in white wine and yeast hulls in red wine were the most effective non-carbonaceous fining agents for the removal of OTA. PMID- 16713648 TI - Diversity of plant cell wall esterases in thermophilic and thermotolerant fungi. AB - Fourteen thermophilic and thermotolerant fungal strains isolated from composting soils produced plant cell wall-acting esterases in a medium containing corn cobs and oat spelt xylan. The concentrated and dialyzed protein extracts of these fungi were fractionated using isoelectric-focusing, gels sliced and eluted protein in each slice was assayed for esterase activity against p-nitrophenyl acetate. A total of 84 esterases detected on the basis of pI were found to show distinct preferential substrate specificities towards p-nitrophenyl acetate, p nitrophenyl ferulate and p-nitrophenyl butyrate, and were putatively classified as acetyl esterases and esterases types I and II. None of the esterases were active against p-nitrophenyl myristate. In addition, these esterases were characterized as acid, neutral or alkaline active. PMID- 16713649 TI - Acid-responsive gene regulation in the human pathogen Helicobacter pylori. AB - Helicobacter pylori is a human gastric pathogen which is extremely well adapted to the low pH environment of the stomach, since it has evolved mechanisms to survive both severe acid shocks and to grow under mildly acidic conditions. Central to the acid resistance of H. pylori is the enzyme urease whose function is to maintain the cytoplasmic and periplasmic pH of the bacterium near neutrality. Substantial progress has been made recently in unravelling the complex regulation of urease expression and the expression of additional genes involved in the acid adaptation of H. pylori. Acid-responsive gene regulation involves the two-component system ArsRS and the metal responsive pleiotropic transcriptional regulators NikR and Fur which control partially overlapping regulons. Here we review our current understanding of the mechanisms of transcriptional regulation governing the acid response of H. pylori. PMID- 16713650 TI - Regioselectivity of 7-O-methyltransferase of poplar to flavones. AB - POMT-7, an O-methyltransferase from poplar (Populus deltoids) was used to modify a variety of flavonoid compounds. POMT-7 was able to transfer a methyl group to several flavonoids containing a C-7 hydroxyl group. However, POMT-7 showed a higher affinity toward flavonol and flavone such as apigenin, kaempferol, luteolin, and quercetin than flavanone and isoflavone. Based on comparison of HPLC retention times with authentic compounds and corresponding nuclear magnetic resonance spectroscopy data, the methylation position of the reaction products was determined to be at the hydroxyl group of C-7. Biotransformation kinetics indicated that the enzyme converted more than 80% of the apigenin, kaempferol, luteolin and quercetin substrates, which were added at concentration of 70 microM, into corresponding 7-methoxy compounds within 24 h. PMID- 16713651 TI - Is there a twenty third amino acid in the genetic code? AB - The universal genetic code includes 20 common amino acids. In addition, selenocysteine (Sec) and pyrrolysine (Pyl), known as the twenty first and twenty second amino acids, are encoded by UGA and UAG, respectively, which are the codons that usually function as stop signals. The discovery of Sec and Pyl suggested that the genetic code could be further expanded by reprogramming stop codons. To search for the putative twenty third amino acid, we employed various tRNA identification programs that scanned 16 archaeal and 130 bacterial genomes for tRNAs with anticodons corresponding to the three stop signals. Our data suggest that the occurrence of additional amino acids that are widely distributed and genetically encoded is unlikely. PMID- 16713652 TI - Genome-wide association: a promising start to a long race. AB - A recent study by Cheung et al. demonstrates how to identify expression quantitative trait loci (eQTLs) underlying gene expression phenotypes through a combination of genome-wide linkage analysis and subsequent fine mapping or by genome-wide association (GWA) analysis. This study emphasizes the complexity of human traits, highlighting the challenges faced by investigators--in particular, insufficient linkage disequilibrium between the trait and marker variant, genetic heterogeneity and correcting for multiple testing will all adversely impact the power to detect loci by association. These issues must be considered carefully if the GWA approach is to succeed in mapping complex phenotypes. PMID- 16713653 TI - Evo-devo and constraints on selection. AB - Developmental bias, or genetic channeling, can influence the tempo and direction of evolution and, thus, become reflected in patterns of biodiversity. Twenty years ago, this notion rested on armchair descriptions of potential constraints on evolution. Now, a broad evo-devo approach involving both evolutionary and developmental genetics provides experimental analysis of such bias, revealing how shared genetic or developmental pathways among morphological traits contribute to the evolution of complexity and diversity, and that developmental bias itself evolves, generating variation in evolvability. As I discuss here, it is now possible to determine the relative importance of natural selection and of genetical and developmental architecture as determinants of organic diversity, particularly for examples of adaptive radiations and parallel evolution. PMID- 16713654 TI - What type of airflow system should be used in orthopaedic operating theatres? PMID- 16713655 TI - MRSA behind bars? PMID- 16713656 TI - Elements of the granular gland peptidome and transcriptome persist in air-dried skin of the South American orange-legged leaf frog, Phyllomedusa hypocondrialis. AB - The defensive strategy of amphibians against predator attack relies heavily on the secretion of noxious/toxic chemical cocktails from specialized skin granular glands. Bioactive peptides constitute a major component of secretions in many species and the most complex are produced by neotropical leaf frogs of the sub family Phyllomedusinae. We recently reported that these skin secretions contain elements of both the granular gland peptidome and transcriptome and that polyadenylated mRNAs constituting the latter are protected from degradation by interactions with endogenous amphipathic peptides. This thus permits parallel amino acid sequencing of peptides and nucleic acid sequencing of cloned precursor transcripts from single lyophilized samples of secretion. Here we report that the protection afforded is sufficiently robust to permit transcriptome studies by cloning of full-length polyadenylated peptide precursor encoding mRNAs from libraries constructed using ambient temperature air-dried skin from recently deceased specimens as source material. The technique was sufficiently sensitive to permit the identification of cDNAs encoding antimicrobial peptides constituted by six different isoforms of phylloseptin and two dermaseptins. Also, for the first time, establishment of the nucleic acid and amino acid sequence of the precursor encoding the phyllomedusine frog skin bradykinin-related peptide, phyllokinin, from cloned cDNA, was achieved. These data unequivocally demonstrate that the granular gland transcriptome persists in air-dried amphibian skin--a finding that may have fundamental implications in the study of archived materials but also in the wider field of molecular biology. PMID- 16713657 TI - Lividins: novel antimicrobial peptide homologs from the skin secretion of the Chinese Large Odorous frog, Rana (Odorrana) livida. Identification by "shotgun" cDNA cloning and sequence analysis. AB - Odorous frogs of the sub-genus Odorrana are of oriental distribution, and are so called due to the foul smell of their defensive skin secretions released from specialized skin glands following stress or predator attack. Here we report the application of a "shotgun" skin secretion cDNA library cloning technique which can rapidly expedite identification of secretion bioactive peptides. From a library constructed from the skin secretion of the Large Chinese Odorous frog, Rana (Odorrana) livida, we have identified four novel peptides whose primary structures were deduced initially from cloned precursors. Subsequently, mature peptides were located in and structurally characterized from reverse phase HPLC fractions of skin secretion. Named lividins 1-4, these were found to be structural homologs of known antimicrobial peptide families from Rana frogs. Rapid identification of novel peptides can thus be rapidly achieved using this non-invasive, non-destructive technology and the extensive similarities revealed between antimicrobial peptide precursor organization and nucleic acid sequences would lend support to the hypothesis that they have a common ancestral origin. PMID- 16713658 TI - The CART receptors: background and recent advances. AB - Previous evidence obtained from several behavioral and biochemical studies suggested the existence of multiple CART receptors. However, identification of CART receptor binding has been largely unsuccessful until recently. The first evidence of CART signaling properties came from a study demonstrating that CART 55-102 inhibited voltage-dependent intracellular calcium signaling. More recent studies showed CART-induced dose- and time-dependent activation of extracellular signal-regulated kinase (ERK) 1 and 2 in AtT20 cell line. The activation of ERK was blocked by pertussis toxin but not genisten suggesting the involvement of Gi/o linked cascade in CART's signaling properties in AtT20 cells. Shortly after these findings, the evidence of CART 61-102 specific binding was obtained from the same cell line. This study demonstrated that [(125)I]-CART 61-102 was displaced only by active CART peptide but not by inactive CART fragments or several other unrelated peptides or drugs. The [(125)I]-CART 61-102 binding was saturable and it had a high affinity for a single site in AtT20 cells. The binding was also dependent on time, pH, temperature and protein concentration. The average (+/-S.E.M.) B(max) and K(d) values were 101.4+/-8.8 fmol/mg protein and 21.9+/-8.0 pM, respectively. These data indicate the existence of specific CART receptor binding in AtT20 cells where CART signaling has been demonstrated. The identification of a receptor clone in these cells may help us elucidate CART receptors in other tissues. Because CART is implicated with several physiological functions including feeding, drug reward and stress, identification of a CART receptor would provide a novel target for the development of pharmacological tools and drugs for obesity and other disorders. PMID- 16713660 TI - GM-CSF-surface-modified B16.F10 melanoma cell vaccine. AB - Through the specific and tight interaction between streptavidin and biotin, a novel platform was developed to allow for rapid (less than 2h), efficient and durable display of streptavidin-tagged bioactive GM-CSF on the surface of biotinylated B16.F10 tumor cells. This technology involved biotinylation of the cell membrane with a biotin derivative and surface modification of the biotinylated cells with the bi-functional fusion protein, streptavidin-tagged GM CSF. Furthermore, the resultant GM-CSF-modified B16.F10 whole tumor cell vaccine could induce strong and long-lasting systemic protection against the wild-type tumor challenge. Therefore, the platform may represent a fast, efficient and safe approach for the whole tumor cell vaccination. PMID- 16713659 TI - Measurement of hippocampal subfields and age-related changes with high resolution MRI at 4T. AB - Histological studies suggest that hippocampal subfields are differently affected by aging and Alzheimer's disease (AD). The aims of this study were: (1) To test if hippocampal subfields can be identified and marked using anatomical landmarks on high resolution MR images obtained on a 4T magnet. (2) To test if age-specific volume changes of subfields can be detected. Forty-two healthy controls (21-85 years) and three AD subjects (76-86 years) were studied with a high resolution T2 weighted fast spin echo sequence. The entorhinal cortex (ERC), subiculum, CA1, CA2 and CA3/4 and dentate were marked. A significant correlation between age and CA1 (r=-0.51, p=0.0002) which was most pronounced in the seventh decade of life was found in healthy controls. In AD subjects, CA1 and subiculum were smaller than in age-matched controls. These preliminary findings suggest that measurement of hippocampal subfields may be helpful to distinguish between normal aging and AD. PMID- 16713661 TI - Immune response to influenza vaccination in community-dwelling Chinese elderly persons. AB - We investigated the immune antibody response to influenza vaccine in community dwelling Chinese elderly persons in Hong Kong. One hundred and twenty-eight subjects were recruited in a single-blind, randomized, and placebo-controlled trial. There was no significant baseline difference between the vaccine and placebo groups regarding the seroprotection rates (PR) (haemagglutination inhibition [HI] titre>or=1:40) and geometric mean titres (GMT) of the HI antibody titers. The PR, GMTs and serological response rates increased significantly in the vaccinated versus placebo groups in A-H1N1 at both weeks 4 and month 6. The GMTs and serological response rates but not the PR for A-H3N2 and influenza B increased significantly in vaccinated versus placebo group at week 4 and month 6 post-vaccination. Multivariate logistic regression analyses of the seroconversion rate for A-H3N2 within the vaccinated group showed that gender, coronary heart disease and the serum albumin level were significant predictors (p=0.018, 0.009 and 0.025, respectively). Influenza vaccination provoked a protective HI antibody response in community-living Chinese elderly persons. The mean number of unplanned hospital admissions per subject over 6 months was significantly lower in the vaccinated than in the placebo groups. Hospitalized elderly persons had poorer nutrition, 4-week post-immunization HI antibody titres and lower mini mental state examination (MMSE) score than non-hospitalized elderly persons. Logistic regression analyses showed that chronic obstructive airway disease significantly increased the risk of hospitalization while the serum albumin level and 4-week A-H3N2 PR (HI>or=40) were independent predictors of a decreased risk of hospitalizations. PMID- 16713662 TI - Natural radioactivity of granites used as building materials. AB - Sixteen kinds of different granites, used as building materials, imported to Greece mainly from Spain and Brazil, were sampled and their natural radioactivity was measured by gamma-ray spectrometry. The activity concentrations of (238)U, (226)Ra, (232)Th and (40)K of granites are presented and compared to those of other building materials as well as other granite types used all over the world. In order to assess the radiological impact from the granites investigated, the absorbed and the effective doses were determined. Although the annual effective dose is higher than the limit of 1mSvy(-1) for some studied granites, they could be used safely as building materials, considering that their contribution in most of the house constructions is very low. An attempt to correlate the relatively high level of natural radioactivity, shown by some of the granites, with their constituent radioactive minerals and their chemical composition, was also made. PMID- 16713663 TI - Formation of compartments in the neck to facilitate suction drainage during transfer of free flaps. AB - A method of surgical compartmentalisation of the neck is reported. This technique can be used in order to maintain suction drainage without compromising the flow of the anastomoses when an end-to-side technique in the internal jugular vein is used. PMID- 16713664 TI - Epiphytic lichen growth abnormalities and element concentrations as early indicators of forest decline. AB - Thallus morphology and element concentrations (S, Al, Fe, Sr, Mn, Ni, Zn, Cu, Pb, As, F, and Cl) were compared in samples of the fruticose lichen genus Usnea at two heights of the Abies sibirica canopy in the East Sayan Mountains (Krasnoyarsk District, Russia) sampled from three stations at 15, 25 and 35 km from Krasnoyark. Usnea species with an abnormal morphology dominated on branches in the upper canopy, 15-22 m above ground level, and normal thalli on lower tree branches, 2-5 m above ground. Abnormal thalli at the tree-top level contained higher Al, Fe, Zn, F, Sr and Pb concentrations compared with normal thalli growing below, confirming a dust impact. No such clear trend was observed between sampling stations. Crown canopy architecture, surface microtopography and the balance between the processes of deposition and the movement and loss of particles play a major role in particle interception and in pollutant delivery to Usnea. PMID- 16713665 TI - Is the diversity of epiphytic lichens a reliable indicator of air pollution? A case study from Italy. AB - This work provided additional information for a better interpretation of lichen diversity values in biomonitoring studies of air pollution. The effects of 12 predictive variables were estimated for the Genova province (NW Italy) by means of a non-parametric model. The diversity of epiphytic lichens was strictly correlated with mean annual rainfall and mean annual temperature. Different variables were found to affect the lichen diversity in urban vs. forested areas. In urban areas, air pollutants, mainly SO2, are still the main limiting factor, even if under ameliorating conditions this relationship becomes less significant. In forested areas, harvesting and forest fires showed a predominant effect, suggesting the need to develop a more defined sampling protocol to estimate atmospheric pollution in such ecosystems. PMID- 16713666 TI - Rethinking the biological clock: eleventh-hour moms, miracle moms and meanings of age-related infertility. AB - Over the past generation, aging and female reproduction have been lodged within the gendered and gendering debates regarding women's involvement in the workforce and demographic shifts toward delayed parenting that culminate in discourses on the "biological clock". Technological solutions to the biological clock, specifically in vitro fertilization, have led to clinical attempts to assess "ovarian reserve", or qualitative and quantitative changes in the ovary that correlate with aging and with successful infertility treatment. Rupturing the longstanding historical connections between menstruation and female reproductive capacity by specifically focusing on the aging of a woman's eggs, the clinical designation of "diminished ovarian reserve" has come to imply that a woman has "old eggs". This is associated in practitioners' and patients' minds with the eclipse of a woman's reproductive potential and with hidden harbingers of menopause. In an ethnographic interview study of 79 couples in the US who conceived after using donor oocytes, we found that women voiced two different narratives that described their experience and attitudes when confronted with an apparent age-related decline in their fertility. The "eleventh-hour mom" narrative was voiced by women who initially tried to become pregnant with their own eggs and turned to donated oocytes as a second-choice option, whereas the "miracle mom" narrative was expressed by women who were generally older, some of whom had entered infertility treatment hoping to conceive with their own eggs, but some who knew from the outset that it was not going to be possible. Through their narratives women not only embodied and made meaningful "diminished ovarian reserve" in varying ways that connect with cultural, social, structural/organizational, symbolic and physical aspects of aging, they reproduced the socio-biological project of the biological clock, but rooted this social project in the metaphor of "old eggs" rather than menopause. PMID- 16713667 TI - Protective effects of N-acetylcysteine treatment post acute paraquat intoxication in rats and in human lung epithelial cells. AB - An animal study in rats and a cell culture study in normal human lung epithelial cells were conducted to evaluate the efficacy of N-acetyl-l-cysteine (NAC) in paraquat intoxication and associated inflammatory and oxidative stress. The effectiveness of post treatment was measured by the change of mortality rates and markers of oxidative stress, including glutathione, malondialdehyde and superoxide anion production. In addition, the levels of nitric oxide were also examined in both animal and cell culture system. NAC treatment does significantly increase the probability of survival in paraquat-intoxicated rats. It can suppress the serum malondialdehyde levels and production of superoxide anions, and conversely, augment total glutathione concentrations in all studying tissues significantly. Moreover, NAC treatment post in paraquat intoxication could reduce destruction of lung tissue, showing less inflammatory cell infiltration in interstitial stroma and mild vascular congestion. The levels of nitrite in serum and BALF were lower than those of the PQ-treated rats. Similarly, levels of iNOS expression and nitrite formation were significantly lower in normal human lung epithelial cells treated with PQ and NAC than PQ-treated alone cells. PMID- 16713668 TI - Postnatal exposure of the male mouse to 2,2',3,3',4,4',5,5',6,6'-decabrominated diphenyl ether: decreased epididymal sperm functions without alterations in DNA content and histology in testis. AB - 2,2',3,3',4,4',5,5',6,6'-Decabrominated diphenyl ether (PBDE 209) is the second most used brominated flame retardant (BFRs) in constructed materials because it is considered less toxic than others, though other fire retardants, some congeners of PBDE 209, are reported to be toxic. This combined the fact that PBDE 209 has been found in high levels in human milk, blood, indoor environments as well as in foodstuffs has led us in this study attempt to find out whether PBDE 209, also known as decaBDE and decabrominated diphenyl oxide (DBDPO), has an adverse effect on this histology of testes and sperm in CD-1 male mice. The mice we studied were divided into groups and gavaged with 10, 100, 500 and 1500 mg/kg PBDE 209 in corn oil per day between postnatal Days 21 and 70. On Day 71, the mice were anesthetized and sperm function, testis DNA content, and histopathology were studied. We found in the 500- and 1500-mg/kg/day groups that neonatal exposure to PBDE 209 reduced sperm epididymal sperm mitochondrial membrane potential (MMP), reduced amplitude of the lateral head displacement (ALH) and induced the generation of hydrogen peroxide (H2O2) in the sperm of sexually mature male mice, without affecting the sperm count, motility, morphology, curvilinear velocity (VCL), angular progressive velocity (VAP), straight-line velocity (VSL), beat-cross frequency (BCF), sperm chromatin structure assay (SCSA), superoxide anion (O2-*) generation, DNA content in testis cells, or testicular histopathology. ALH was positively associated with an increase in MMP and negatively associated with generation of sperm H2O2. The reduction of MMP was negatively associated with an increase in generation of sperm H2O2. The presence of the relationships between sperm ALH, MMP, and generation of H2O2 indicate toxic action possibly resulting from PBDE 209-induced oxidative stress. In conclusion, this is the first study to report the lowest-observed-adverse-effect level (LOAEL) for sperm function to be 500 mg/kg of PBDE 209 in male mice. Decreased epididymal sperm MMP and ALH as well as induced generation of sperm H2O2 were some of the most serious effects of postnatal PBDE 209 exposure. Future investigations should be performed to study the effects of prenatal exposure of PBDE 209 and the mechanism behind PBDE 209-related oxidative stress in the fetal and pubertal stages of development. PMID- 16713669 TI - Impact of the acid flow rate on dentin erosion. AB - OBJECTIVE: The study aimed to evaluate the erosive effects of acids flowing with different velocities and duration on dentin previously pre-treated with distilled water or human saliva. METHODS: Bovine dentin surfaces were submitted to a 10 min demineralisation with hydrochloric or citric acid (pH 2.3, 37 degrees C) in an artificial mouth at flow rates of 3, 2.25, 1.5, 0.75 or 0.15 ml/min or in 30 ml of the respective acid without movement (each subgroup n=16 specimens). Prior to the demineralisation, half of the specimens of each group were either pre-treated with distilled water or human saliva for 120 min. After 2, 5 and 10 min demineralisation, profilometric traces of dentin loss were performed perpendicular to the direction of acid flow at three sites (A-C) with intervals of 500 microm. The first profile (A) was taken in a distance of 1.5 mm from the acid starting to flow over the sample. Thus, median dentin loss at each site was calculated and statistically analysed by five-way ANOVA. RESULTS: Dentin loss increased with increasing acid flow rate and demineralisation time and was higher for demineralisation with citric acid compared to hydrochloric acid. For given flow rates of 0.15 to 3 ml/min, measurement of dentin loss yielded highest values for site A and decreasing values for sites B and C. For all groups, no significant differences of dentin loss were observed between specimens pre treated with distilled water or human saliva. CONCLUSION: Under the conditions of this study, both duration of demineralisation and acid flow rate but not pre treatment with distilled water or saliva influence the extent of dentin erosion. PMID- 16713670 TI - Plasminogen activator inhibitor-1 expression and secretion are stimulated by growth hormone and interleukin-6 in 3T3-L1 adipocytes. AB - Various adipocytokines have been described which influence insulin sensitivity and vascular function profoundly and might, therefore, potentially link obesity, insulin resistance, and atherosclerosis. Among those, plasminogen activator inhibitor (PAI)-1 is an adipose-secreted factor upregulated in obesity and insulin resistance that inhibits fibrinolysis. Furthermore, recent studies in knockout mice suggest that PAI-1 directly impairs insulin sensitivity. In the current study, the impact of growth hormone (GH) and interleukin (IL)-6 on PAI-1 mRNA synthesis and secretion was determined in 3T3-L1 adipocytes. Interestingly, 500 ng/ml GH and 30 ng/ml IL-6 increased PAI-1 secretion five-fold and 3.6-fold, respectively. Furthermore, GH and IL-6 induced PAI-1 mRNA by up to 7.3-fold, and 3.6-fold, respectively, in a time-dependent fashion with significant stimulation seen at concentrations as low as 5 ng/ml GH and 10 ng/ml IL-6. Other insulin resistance-inducing hormones which stimulated PAI-1 synthesis included insulin, TNFalpha, and dexamethasone. Studies using pharmacological inhibitors suggested that basal and GH-induced PAI-1 synthesis were at least in part mediated by p44/42 mitogen-activated protein kinase but not janus kinase 2 and phosphatidylinositol 3-kinase. Taken together, our results show a differential regulation of PAI-1 mRNA by insulin resistance-inducing hormones including GH and IL-6. PMID- 16713671 TI - Epidemiology of myotonic dystrophy type 1 (Steinert disease) in Belgrade (Serbia). AB - The aim of this study was to estimate the incidence and prevalence of myotonic dystrophy type 1 (DM1) in Belgrade during the period 1983-2002. The patients who had DM1 were ascertained through hospital records from all neurological departments in Belgrade during 1983-2002. The molecular genetic analysis was performed in all patents included in the study. We identified 101 DM1 patients (52 males and 49 females). The average annual incidence rate of DM1 in Belgrade for the period observed was 2.0/1,000,000 (95% confidence interval (CI), 0.3 8.3), 2.1/1,000,000 (95% CI, 0.3-8.3) for males and 2.0/1,000,000 (95% CI, 0.3 8.3) for females. The highest age-specific DM1 incidence was registered in the age group 20-49: 3.4/1,000,000 (95% CI, 0.5-7.6), 4.0/1,000,000 (95% CI, 1.1 10.2) in males and 2.5/1,000,000 (95% CI, 0.5-7.6) in females. In the population of Belgrade, a cumulative probability of acquiring DM1 was 1 per 8621 for men and 1 per 9259 for women (1 per 8940 of the population for both sexes). The prevalence of DM1 in Belgrade on 31 December 2002 was 5.3/100,000 (95% CI, 4.2 6.6). PMID- 16713672 TI - Multistage carcinogenesis in Barrett's esophagus. AB - The multistage carcinogenesis of esophageal adenocarcinoma is a process of clonal evolution within Barrett's esophagus neoplasms. The initiating event for Barrett's esophagus is unknown, but is associated with chronic gastric reflux which probably also promotes progression. Inactivation of both alleles of CDKN2A appear to be early events causing clonal expansion. Clones with TP53 inactivated expand if they have already inactivated CDKN2A. After TP53 has been inactivated, tetraploid and aneuploid clones tend to develop. The final events that lead to invasion and metastasis are unknown. Evolutionary biology provides important tools to understand clonal evolution in progression and cancer prevention. PMID- 16713673 TI - Variations in the peroxisome proliferator-activated receptor-gamma gene and melanoma risk. AB - There is strong evidence to suggest that the peroxisome proliferator-activated receptor (PPAR)-gamma, a member of the nuclear receptor family of transcriptional regulators, mediates tumor suppressive activities in a variety of human cancers. Recently, PPARgamma agonists were found to inhibit growth of melanoma cell lines. Here, we tested the possibility that variations in the gene encoding PPARgamma (PPARG) influence melanoma risk. Two variations of PPARG (P12A[rs1801282] and C161T [rs3856806]) were investigated in two independent case-control studies with a total of 832 melanoma patients and 790 control individuals. In the first study, homozygous carriers of the rare *T allele of the C161T polymorphism in exon 6 of PPARG were significantly more common among patients with melanoma than among healthy subjects (6.0 vs. 2.0%; P=0.0096) and this association was independent of clinical risk factors such as skin type and nevus count (odds ratio 5.18; 95% confidence interval 1.68-15.96; P=0.0041). This finding, however, could not be replicated in the second case-control study. We therefore conclude that the investigated PPARG polymorphisms are not likely to constitute a significant risk factor for the development of melanoma among German Caucasians. PMID- 16713674 TI - IR microspectroscopy: potential applications in cervical cancer screening. AB - Screening exfoliative cytology for early dysplastic cells reduces incidence and mortality from squamous carcinoma of the cervix. In the developed world, screening programmes have adopted a 3-5 years recall system. In its absence, cervical cancer would be the second most common female cancer in these regions; instead, it is currently eleventh. However, there exist a number of limitations to the smear test even given the removal of contaminants using liquid-based cytology. It is prohibitively expensive, labour-intensive and subject to inaccuracies that give rise to significant numbers of false negatives. There remains a need for novel approaches to allow efficient and objective interrogation of exfoliative cytology. Methods that variously exploit infrared (IR) microspectroscopy are one possibility. Using IR microspectroscopy, an integrated 'biochemical-cell fingerprint' of the lipid, protein and carbohydrate composition of a biomolecular entity may be derived in the form of a spectrum via vibrational transitions of individual chemical bonds. Powerful statistical approaches (e.g. principal component analysis) now facilitate the interrogation of large amounts of spectroscopic data to allow the extraction of what may be small but extremely significant biomarker differences between disease-free and pre-malignant or malignant samples. An increasing wealth of literature points to the ability of IR microspectroscopy to allow the segregation of cells based on their disease status. We review the current evidence supporting its diagnostic potential in cancer biology. PMID- 16713675 TI - Degradation of di-n-butyl phthalate using photoreactor packed with TiO2 immobilized on glass beads. AB - This study evaluated the performance of a photoreactor packed with TiO2/glass, TiO2 immobilized on glass beads, initiated by UV irradiation, denoted as UV/TiO2/glass, to decompose di-n-butyl phthalate (DBP) in an aqueous solution. The photodegradation rate of DBP by this UV/TiO2/glass process was found to obey pseudo first-order kinetics represented by the Langmuir-Hinshelwood model. The experimental results of this study show that the influence of pH value of an aqueous solution to reaction rate was negligible at the pH values 4.5-9. The effect of cations on the photodegradation rate of DBP reveals that the larger the charge and size of cations contained, the more the inhibition of reaction rate increased. The UV/TiO2/glass process yielded a 75% degradation efficiency of DBP with initial concentration of 5 mg L(-1) at 80 min reaction time. PMID- 16713676 TI - Effects of quetiapine on the brain-derived neurotrophic factor expression in the hippocampus and neocortex of rats. AB - The effects of antipsychotics on the brain-derived neurotrophic factor (BDNF) expression have been controversial. This study aimed to investigate the effects of chronic quetiapine administration on the BDNF mRNA expression in hippocampus and neocortex of rats with or without immobilization stress. The chronic administration (21 days) of quetiapine (10 mg/kg) significantly attenuated the decreased BDNF mRNA expression in the both hippocampal and cortical regions of rats caused by immobilization stress, and significantly increased the BDNF mRNA expression in the dentate gyrus of rats even without the immobilization stress. These results could add some theoretical bases to explain why quetiapine may improve cognitive symptoms of schizophrenia by stimulating BDNF mRNA expression. PMID- 16713677 TI - Bone marrow stromal cells can achieve cure of chronic paraplegic rats: functional and morphological outcome one year after transplantation. AB - Chronic paraplegia resulting from severe spinal cord injury (SCI) is considered to be an irreversible condition. Nevertheless, recent studies utilizing adult stem cells appear to offer promise in the treatment of this and other neurological diseases. Here, we show that progressive functional motor recovery is achieved over the course of the year following the administration of bone marrow stromal cells (BMSC) in traumatic central spinal cord cavities of adult rats with chronic paraplegia. At this time, functional recovery is almost complete and associated with evident nervous tissue regeneration in the previously injured spinal cord. PMID- 16713678 TI - High resolution non-contact AFM imaging of liquids condensed onto chemically nanopatterned surfaces. AB - The wetting of ethanol and octane on chemically nanopatterned surfaces has been investigated using Atomic Force Microscopy (AFM) under controlled environmental conditions. The patterns were generated on a methyl-terminated, organic monolayer using an AFM electro-oxidation process. The subsequent wetting of the organic liquids was studied using non-contact mode AFM under equilibrium conditions with the vapor. This study of condensed nanoliquids provides the first reliable measurements of sub 100 nm liquid profile shapes. The derived contact angles give an estimate of the line tension. PMID- 16713679 TI - Effect of biological control through the daily application of spores of Duddingtonia flagrans in lambs kept under an evasive grazing system in the Netherlands. AB - In 2004, an experiment was carried out to evaluate the effect of biological control through feeding spores of Duddingtonia flagrans on parasitic gastroenteritis in lambs, kept under an evasive grazing system. In total 66 lambs were used. Forty naturally infected 3-month old ram lambs were weaned in mid June, and divided into four groups of 10 lambs. On 21 June, G1-G4 were moved to four separate virtually clean plots, they were moved after 4 and 8 weeks to similar plots, and housed after 12 weeks to be necropsied 16 days later. The other 26 lambs had been raised helminth-free, and were used as pairs of tracer lambs. All but one of these pairs, were grazed during the last 2 weeks on each plot. The remaining pair (TA) was grazed during the last 2 weeks on pasture (30 August to 13 September) on the plot that had been grazed by G3 between 19 July and 16 August, to study inhibited development in Haemonchus contortus. All lambs were fed 200 g of concentrates daily throughout the whole period, and those of G1 and G2 were also fed 500,000 spores of D. flagrans/kg bodyweight daily. The faecal cultures demonstrated a high reduction in yield as a result of fungal application. However, no differences between groups were seen in weight gain, faecal egg counts, pasture larval counts, worm counts and tracer worm counts. H. contortus was the dominant species, and it is obvious that the moves at 4-week intervals prevented the development of severe haemonchosis. This is in particular demonstrated by the much higher worm counts in the two TA tracer lambs grazed. Nevertheless, increases to high faecal egg counts 3 weeks after the first and second moves, indicated acquisition of infection before these moves and at least subclinical haemonchosis. This was supported with the worm counts of lambs and tracer lambs. A higher proportion of inhibited early L4 than in other tracers and than in the permanent lambs were found in the pair of TA tracer lambs. This indicates that moves to new pastures in late summer and autumn delays the onset of inhibition. PMID- 16713680 TI - Hyaluronidase and CD44 hyaluronan receptor expression in squamous cell laryngeal carcinoma. AB - Squamous cell laryngeal carcinoma undergoes significant structural-related modifications of the extracellular matrix components (ECM), the most characteristics being the presence of degraded collagen, aggrecan and hyaluronan. We examined the presence of hyaluronidase and of the cellular hyaluronan receptor CD44 during the various stages of cancer. ECM components were extracted by using PBS, 4 M GdnHCl and 4 M GdnHCl-0.1% Triton-X 100 sequentially and hyaluronidase and CD44 analyzed by zymography and immunochemistry techniques. Total RNA was also extracted and the mRNA of the various hyaluronidases and of CD44 was analyzed after amplification with RT-PCR. Hyaluronidase was detected as a double band of 45 and 55 kDa molecular mass, only in cancer samples. The analysis of mRNA indicated an aberrant expression of PH-20, the testicular-type hyaluronidase, at late stages of cancer and an overexpression of HYAL1 only at stage IV. In addition, CD44 was identified in two protein bands of 80 and 64 kDa in cancer samples. The analysis of mRNA showed that hyaluronan receptor was expressed in a stage-related order. Thus, it could be suggested that in laryngeal squamous cell carcinoma, cancer cells migrated and proliferated under the influence of small molecular mass hyaluronan, by expressing increased amounts of its receptor. PMID- 16713681 TI - A globotriaosylceramide (Gb3Cer) mimic peptide isolated from phage display library expressed strong neutralization to Shiga toxins. AB - A Gb3-trisaccharide mimic peptide was selected with biopanning from a phage display library against anti-Gb3 antibody to neutralize Shiga toxins (Stxs). Biopanning was carried out on a microplate immobilized with a Fab fragment of anti-Gb3 antibody and a subtraction procedure screening was applied to enhance specificity. The selected phage clones showed strong affinity to anti-Gb3 antibody and to Stxs. Among these clones, a 9-mer sequence WHWTWLSEY was determined as the strongest Gb3 mimic peptide and chemically synthesized. The peptide bound strongly to Stx-1 and Stx-2, though the binding was inhibited with Gb3Cer. Surface plasmon resonance (SPR) and fluorescent spectroscopy determined that the affinity of the peptide to both Stxs was strong. Neutralization activity was confirmed by in vitro assay with HeLa cells. The Gb3 mimic peptide potentially has great promise for use against Stxs. PMID- 16713682 TI - Brain and cognitive impairments from burn injury in rats. AB - Severe burn injuries affect not only the release of stress hormones but also the metabolism of nitric oxide (NO), a substance playing a large role in cognition. We investigated the effect of third-degree burns both on central NO-levels and on short-term memory in rats. Burns were administered under halothane-anesthesia by dipping 20% of the skin area in hot water. In a first experiment, NO-changes were estimated over hours by differential normal pulse voltammetry (DPNV) with a sensor implanted chronically in the frontal cortex. In a second experiment, cognitive abilities were tested over days by comparing the spontaneous time used to explore objects that the animals had, either never- or already-encountered before. Cerebral NO appeared steadily depleted for at least 12h after the injury, not after control anesthesia. During nine days following the burn, discrimination performance was lower compared to controls. Putting together the results of the two experiments, especially on the day of burn, NO changes are likely to account for the behavioral effect. A choice of neuro-pharmacological agents involved in NO-metabolism, together with a choice of proper anesthetics, should now be tested as means to alleviate cognitive impairments following third-degree burns. PMID- 16713683 TI - Paediatric burn injuries in Sub Saharan Africa--an overview. AB - Paediatric burn injuries in Sub Saharan Africa are common and often lead to devastating consequences. Unfortunately relevant and accurate data regarding these injuries is sketchy and incomplete. This paper reviews the available information on the epidemiology of paediatric burns in Africa, associated health problems and contributing environmental factors responsible for these burns. The current status of burn care, the lack of infrastructure, and traditional methods of treatment, further contribute to the unsatisfactory status of overall burn management, prevention, and rehabilitation of burn survivors. A strategy for improving burn care in Africa has been formulated. The management of childhood burns will only be successful if educational, social, fiscal and infrastructure standards are improved. Traditional beliefs and methods cannot be discarded as they play an important role in the management of these children. It is furthermore essential that local and central government organisations support these initiatives. Clearly, the children of Africa deserve better burn care. PMID- 16713684 TI - Quantification of mentolabial profile using lateral roentgen cephalography and profile reconstruction by mandibular osteotomy in healthy adults and patients with scar contracture following severe facial burns. AB - Here mentolabial form including lateral facial contour and bony contour in healthy adults was studied to refer to values of normal mentolabial form using lateral roentgen cephalography. In soft tissue of healthy adults, thickness of mentolabial soft tissue was greater for men than for women. Degree of mentolabial curvature was greater for men than for women. In severe facial burn patients, the micrognathia-like deformity was evaluated compared with the reference values. In soft tissue of severe facial burn patients, loss of mentolabial sulcus and eversion of the lower lip were characteristically analyzed, suspecting that the major cause was scar contracture. These statistical data contribute to reconstruction of micrognathia-like deformity after severe facial burn to lead to favorable mentolabial form. It is shown that osteotomized genio-advancement produced ideal mentolabial form in severe facial burn patients, who had not been satisfied with the form by conventional operative methods, free skin flap, skin flap or free skin graft. And that the analyzed data, especially in the parameter of thickness of mentolabial sulcus, was improved. PMID- 16713685 TI - Characteristics of 1494 pediatric burn patients in Shanghai. AB - To analyze the epidemiological characteristics of pediatric burn patients in Shanghai and to determine the targets for a pediatric burn prevention program, a retrospective review of all medical records of acute pediatric burn patients (age 0.1) between fertile and subfertile stallions. However, the molar ratio of cholesterol-to-phospholipid was 2.5 times greater in the seminal plasma (P = 0.09) and 1.9 times greater (P = 0.009) in whole sperm of subfertile stallions compared to fertile stallions. PMID- 16713690 TI - Microprocessor controlled transdermal drug delivery. AB - Transdermal drug delivery via iontophoresis is reviewed with special focus on the delivery of lidocaine for local anesthesia and fentanyl for patient controlled acute therapy such as postoperative pain. The role of the microprocessor controller in achieving dosimetry, alternating/reverse polarity, pre-programmed, and sensor-based delivery is highlighted. Unique features such as the use of tactile signaling, telemetry control, and pulsatile waveforms in iontophoretic drug delivery are described briefly. PMID- 16713691 TI - Novel polymeric micelles for hydrophobic drug delivery based on biodegradable poly(hexyl-substituted lactides). AB - Novel amphiphilic methoxy-poly(ethylene glycol)-poly(hexyl-substituted lactides) block copolymers were synthesized by ring-opening polymerization (ROP) of mono and dihexyl-substituted lactide (mHLA and diHLA) in bulk at 100 degrees C in the presence of tin(II) 2-ethylhexanoate (Sn(Oct)(2)) as catalyst and methoxy poly(ethylene glycol) (MPEG) as initiator. MPEG-PmHLA and MPEG-PdiHLA copolymers of predictable molecular weights and narrow polydispersities were obtained, as shown by (1)H NMR and GPC. DSC experiments showed that the MPEG-PHLA block copolymer presents a bulk microstructure containing MPEG domains segregated from the PHLA domains. Micelles were successfully prepared from these block copolymers, with sizes ranging from 30 to 80 nm. The critical micellar concentration (CMC) was found to decrease with the increasing number of hexyl groups on the polyester block (MPEG-PLA > MPEG-PmHLA > MPEG-PdiHLA) for copolymers of the same composition and molecular weight. The hydrophobicity of the micelle core in dependence of the number of hexyl groups along the PLA chain was evidenced by absorbance experiments with the incorporation of the dye Nile Red. These novel amphiphilic copolymers are interesting for micellar drug delivery and especially in regard to optimized hydrophobic drug loadings, as it was shown for griseofulvin as a model drug. PMID- 16713692 TI - Potassium currents in human myogenic cells from donors of different ages. AB - Ageing in humans is accompanied by a reduction in the capacity of satellite cells to proliferate and the forming myoblasts to fuse. The processes of myoblast differentiation and fusion are associated with specific changes in the cells electrical properties. We wanted to elucidate the possible effects of ageing on these parameters and performed whole-cell patch-clamp recordings on human myoblasts obtained from biopsies of skeletal muscles from 2-, 48- and 76-year-old donors. First, we found that resting membrane potential on the 4th day of differentiation in vitro is less negative in the older than in the younger cells. Moreover, the oldest cells showed a smaller density of outward and inward potassium currents. More cells from the old and middle-age donors have a low (less than -40 mV) potential of activation for the outward potassium current. We conclude that in human myoblasts biophysical properties of potassium currents change with donor age. PMID- 16713693 TI - Sustained inhibition of oxidative phosphorylation impairs cell proliferation and induces premature senescence in human fibroblasts. AB - The mitochondrial theory of aging predicts that functional alterations in mitochondria contribute to the aging process. Whereas this hypothesis implicates increased production of reactive oxygen species (ROS) as a driving force of the aging process, little is known about molecular mechanisms by which mitochondrial impairment might contribute to aging. Using cellular senescence as a model for human aging, we have recently reported partial uncoupling of the respiratory chain in senescent human fibroblasts. In the present communication, we address a potential cause-effect relationship between mitochondrial impairment and the appearance of a senescence-like phenotype in young cells. We found that treatment by antimycin A delays proliferation and induces premature senescence in a subset of the cells, associated with increased reactive oxygen species (ROS) production. Quenching of ROS by antioxidants did however not restore proliferation capacity nor prevent premature senescence. Premature senescence is also induced upon chronic exposure to oligomycin, irrespective of ROS production, and oligomycin treatment induced the up-regulation of the cdk inhibitors p16, p21 and p27, which are also up-regulated in replicative senescence. Thus, besides the well established influence of ROS on proliferation and senescence, a reduction in the level of oxidative phosphorylation is causally related to reduced cell proliferation and the induction of premature senescence. PMID- 16713694 TI - Social insects as a model to study the molecular basis of ageing. AB - One major gap in the current knowledge of the molecular bases of ageing is that most of the work has been done using short-lived model organisms such as fruitflies, nematodes, yeast and mice. Here, we argue that ants and social bee species provide an excellent complementary system to study ageing, and this for two reasons: first, in contrast to model organisms, ant and bee queens are extraordinarily long-lived, and second, there is a tremendous variation in lifespan among the genetically identical queens, workers (non-reproductive females) and males, with queens living up to 500 times longer than males and 10 times longer than workers. We review recent experimental work aimed at testing the role of antioxidant genes within the conceptual framework of the free radical theory of ageing, as well as studies investigating the role of juvenile hormone, vitellogenin and telomeres as mediators of ageing in social insects. PMID- 16713695 TI - Near-real-time feedback control system for liver thermal ablations based on self referenced temperature imaging. AB - Our challenge was to design and implement a dedicated temperature imaging feedback control system to guide and assist in a thermal liver ablation procedure in a double-donut 0.5T open MR scanner. This system has near-real-time feedback capability based on a newly developed "self-referenced" temperature imaging method using "moving-slab" and complex-field-fitting techniques. Two phantom validation studies and one ex vivo experiment were performed to compare the newly developed self-referenced method with the conventional subtraction method and evaluate the ability of the feedback control system in the same MR scanner. The near-real-time feedback system was achieved by integrating the following primary functions: (1) imaging of the moving organ temperature; (2) on-line needle tip tracking; (3) automatic turn-on/off the heating devices; (4) a Windows operating system-based novel user-interfaces. In the first part of the validation studies, microwave heating was applied in an agar phantom using a fast spoiled gradient recalled echo in a steady state sequence. In the second part of the validation and ex vivo study, target visualization, treatment planning and monitoring, and temperature and thermal dose visualization with the graphical user interface of the thermal ablation software were demonstrated. Furthermore, MR imaging with the "self-referenced" temperature imaging method has the ability to localize the hot spot in the heated region and measure temperature elevation during the experiment. In conclusion, we have demonstrated an interactively controllable feedback control system that offers a new method for the guidance of liver thermal ablation procedures, as well as improving the ability to assist ablation procedures in an open MR scanner. PMID- 16713696 TI - Chemical and biological characterisation of nutraceutical compounds of broccoli. AB - People's diet offers a greater and more diverse group of plant bioactives than do drugs, and they often do not realise that many drugs are derived from the compounds originally discovered in plant foods. Numerous epidemiological studies indicate that Brassica vegetables in general, and broccoli in particular, protect humans against cancer since they are rich sources of glucosinolates as well as possessing a high content of flavonoids, vitamins and mineral nutrients. One unusual phytotherapeutic role of broccoli is for skin diseases--the juice of the leaves is used to treat warts. However, the main use of broccoli stems from its health-promoting properties. Some criteria have been proposed to evaluate the possibilities of developing new "functional foods" to reduce the risk of specific cancers; largely in broccoli, which is associated with cancer protection. Processing conditions, transport, domestic cooking, etc., affect the health promoting properties of broccoli and these have been widely studied. This review makes an in-depth study of the chemical and biological characterization of the phytochemicals of broccoli and the effects on the bioactive composition of broccoli. PMID- 16713697 TI - Sensitive bioassay for the simultaneous determination of pseudoephedrine and cetirizine in human plasma by liquid-chromatography-ion trap spectrometry. AB - A liquid chromatography-ion trap mass spectrometry coupled with electrospray ionization (HPLC-ESI-ion trap mass spectrometry) method for simultaneous determination of cetirizine and pseudoephedrine in human plasma is presented. Chromatographic separation was performed on a Hypurity C18 column (Thermo Hypersil-Keystone 2.1 mm x 150 mm, 5 microm, USA), The mobile phase was composed of 65% methanol and 35% water (contained 0.1% formic acid, 10 mM ammonium formate), which was run with a flow-rate of 0.2 ml/min at 40 degrees C. Quantitation was achieved by monitoring the product ions at m/z 166-->m/z 148 (pseudoephedrine), m/z 389.9-->m/z 201.1 (cetirizine), m/z 264-->m/z 246 (tramadol, IS). The calibration curve of pseudoephedrine and cetirizine was established with standard solutions. The limit of detection for pseudoephedrine and cetirizine each was 5 ng/ml. This simplified analytical method is sensitive, specific and accurate enough for simultaneous determination of pseudoephedrine and cetirizine in human plasma and is successfully applied to the pharmacokinetic study of pseudoephedrine and cetirizine. PMID- 16713698 TI - PCB 50 stimulates release of arachidonic acid and prostaglandins from late gestation rat amnion fibroblast cells. AB - Amniotic phospholipase A2 activity contributes to elevated levels of arachidonic acid and prostaglandins observed during labor. Polychlorinated biphenyls (PCBs) activate PLA2 and have been associated with shortened gestation length. To determine if PCBs stimulate amniotic PLA2, cell cultures of rat amnion fibroblasts (RAF) were established from gestation day (gd) 20 rats and labeled with 0.5 micro Ci [3H]-arachidonic acid prior to a 0.5- or 4-h exposure to 0.1% DMSO (solvent control), PCB 50 (1-50 micro M) or TNFalpha (positive control). PCB 50 and TNFalpha induced significant release of [3H]-arachidonic acid from amnion fibroblast cells in time-dependent manners (p<0.001), an effect associated with a significant increase in iPLA2 expression (p<0.05). PCB 50 also stimulated prostaglandin production from RAF cells independent of changes in immunoreactive COX-2. These data suggest that amnion may serve as a target for PCB-induced release of arachidonic acid and uterotonic prostaglandins, with a potential for adverse pregnancy outcomes. PMID- 16713699 TI - Antiplasmodial and antitrypanosomal activity of new esters and ethers of 4 dialkylaminobicyclo[2.2.2]octan-2-ols. AB - Only three drugs are available for the treatment of East African trypanosomiasis. Patients suffer from painful application, severe side effects and increasing resistance against these drugs. Malaria tropica kills more than 2 million people every year mainly due to growing drug resistance. 4 Dialkylaminobicyclo[2.2.2]octan-2-ols and some of their esters have shown activity against both the causative organisms, Trypanosoma brucei rhodesiense and Plasmodium falciparum. Ethers and new esters with markedly higher lipophilicity were prepared in three-step procedures from acyclic synthons. The new compounds were screened for their antiprotozoal activities against T. b. rhodesiense (STIB 900) and P. falciparum K1 (resistant to chloroquine and pyrimethamine), and for their cytotoxicity with L-6 cells by means of in vitro microplate assays. The results were compared to those of the parent compounds indicating that higher lipophilicity increases the antiprotozoal activities. The pivalate 10a showed the highest antitrypanosomal activity. The 4-chlorobenzoate 9b exhibited good antiplasmodial activity and low cytotoxicity. The most active antiplasmodial agent was the benzhydryl ether 13c which was nearly as active as chloroquine against sensitive strains. PMID- 16713700 TI - Effects of controlled-release on the pharmacokinetics and absorption characteristics of a compound undergoing intestinal efflux in humans. AB - OBJECTIVE: The number of active pharmaceutical ingredients (API) undergoing inhibitable and saturable intestinal efflux is considerable. As a consequence, absorption and bioavailability may depend on the intestinal concentration profile of the drug and may vary as a function of dose and release rate of the drug from the dosage form. The impact of controlled versus immediate-release on the absorption of P-glycoprotein substrates is currently unknown. Thus, the main focus of the present study was a comparison of the pharmacokinetics of the P-gp model substrate talinolol following administration of immediate-release (IR) and controlled-release (CR) tablets to healthy human volunteers with a particular focus on the absorption characteristics of the active pharmaceutical ingredients. METHODS: Talinolol immediate-release (Cordanum), 100mg), one controlled-release (100mg) and two controlled-release tablets (200mg) were administered as single doses to fasting healthy volunteers in a crossover design with a 1 week washout period between treatments. Sufficient blood and urine samples were drawn and analysed using a specific HPLC method with UV detection to describe the resulting plasma and urinary excretion versus time profiles. RESULTS: The bioavailability of talinolol in term of AUC(0-->infinity) for IR talinolol was approximately twice as high as compared to the administration of the same dose in a controlled release dosage form. After administration of talinolol IR tablets, the drug was rapidly absorbed and reached maximum concentrations C(max) of 204.5 ng/ml+/-121.8 (means+/-S.D.) 2h after dosing. The terminal half-life of the drug averaged 19.8h following IR administration in comparison to 32 h under CR dosing conditions. Following administration of the IR dosage form, significant secondary peaks were observed in one healthy subject. Secondary peaks were not clearly apparent in the CR plasma profiles. CONCLUSION: The present study demonstrates a considerable loss of bioavailability of drugs that are substrates of intestinal secretory transporters upon their administration in controlled-release dosage forms. PMID- 16713701 TI - In vitro evaluation of N-methyl amide tripeptidomimetics as substrates for the human intestinal di-/tri-peptide transporter hPEPT1. AB - Oral absorption of tripeptides is generally mediated by the human intestinal di /tri-peptide transporter, hPEPT1. However, the bioavailability of tripeptides is often limited due to degradation in the GI-tract by various peptidases. The aim of the present study was to evaluate the general application of N-methyl amide bioisosteres as peptide bond replacements in tripeptides in order to decrease degradation by peptidases and yet retain affinity for and transport via hPEPT1. Seven structurally diverse N-methyl amide tripeptidomimetics were selected based on a principal component analysis of structural properties of 6859 N-methyl amide tripeptidomimetics. In vitro extracellular degradation of the selected tripeptidomimetics as well as affinity for and transepithelial transport via hPEPT1 were investigated in Caco-2 cells. Decreased apparent degradation was observed for all tripeptidomimetics compared to the corresponding natural tripeptides. However, affinity for and transepithelial transport via hPEPT1 were only seen for Gly-Sar-Sar, AsnPsi[CONCH(3)]PhePsi[CONCH(3)]Trp, and Gly-Sar-Leu. This implies that tripeptidomimetics originating from tripeptides with neutral side chains are more likely to be substrates for hPEPT1 than tripeptidomimetics with charged side chains. The results of the present study indicate that the N methyl amide peptide bond replacement approach for increasing bioavailability of tripeptidomimetic drug candidates is not generally applicable to all tripeptides. Nevertheless, retained affinity for and transport via hPEPT1 were shown for three of the evaluated N-methyl amide tripeptidomimetics. PMID- 16713702 TI - The minipig in dermatotoxicology: methods and challenges. AB - In view of more morphological and physiological similarities between human and porcine skin than for other laboratory animal species, the minipig is a preferred model to evaluate the safety profile of dermally applied xenobiotics. Different methods of dermal administration and examples of non-invasive and invasive investigations during the in-life phase of the studies are described. Routine and special post-mortem examinations in dermal studies are presented to assess responses to the topical treatment of minipig skin. Challenges in dermal minipig studies are discussed with respect to animal welfare and husbandry, test formulations, application methods and different types of investigations. One of the most significant issues for dermal minipig studies is the extensive measures required to prevent cross-contamination of blood and tissue samples taken to monitor local and systemic exposure to the test item. PMID- 16713705 TI - Protein homeostasis in neurons and its pathological alterations. AB - In neuronal cells, proteins are synthesized on ribosomes from the genetic information encoded in DNA. In some instances translation takes place at the neuronal cell soma but in other it occurs at distal location, such as in a dendritic spine. Folding is usually initiated before the completion of protein synthesis and its outcome strictly depends on the local environment in which the nascent protein is submerged. Incompletely folded proteins and, more importantly, misfolded proteins are under the surveillance of several quality control systems that re-establish the correct conformation or initiate protein degradation. Regulation and maintenance of these systems is a vital issue for neuronal and glial cells, and impairments at different levels leads to neurodegenerative diseases. PMID- 16713706 TI - Prediction of RNA secondary structure by free energy minimization. AB - RNA secondary structure is often predicted from sequence by free energy minimization. Over the past two years, advances have been made in the estimation of folding free energy change, the mapping of secondary structure and the implementation of computer programs for structure prediction. The trends in computer program development are: efficient use of experimental mapping of structures to constrain structure prediction; use of statistical mechanics to improve the fidelity of structure prediction; inclusion of pseudoknots in secondary structure prediction; and use of two or more homologous sequences to find a common structure. PMID- 16713709 TI - Sequence comparison and protein structure prediction. AB - Sequence comparison is a major step in the prediction of protein structure from existing templates in the Protein Data Bank. The identification of potentially remote homologues to be used as templates for modeling target sequences of unknown structure and their accurate alignment remain challenges, despite many years of study. The most recent advances have been in combining as many sources of information as possible--including amino acid variation in the form of profiles or hidden Markov models for both the target and template families, known and predicted secondary structures of the template and target, respectively, the combination of structure alignment for distant homologues and sequence alignment for close homologues to build better profiles, and the anchoring of certain regions of the alignment based on existing biological data. Newer technologies have been applied to the problem, including the use of support vector machines to tackle the fold classification problem for a target sequence and the alignment of hidden Markov models. Finally, using the consensus of many fold recognition methods, whether based on profile-profile alignments, threading or other approaches, continues to be one of the most successful strategies for both recognition and alignment of remote homologues. Although there is still room for improvement in identification and alignment methods, additional progress may come from model building and refinement methods that can compensate for large structural changes between remotely related targets and templates, as well as for regions of misalignment. PMID- 16713710 TI - A comparison of muscle activity and heart rate response during backward and forward walking on an underwater treadmill. AB - This investigation compared muscle activities and heart rate (HR) responses while subjects walked backward or forward in water, with and without a water current. Ten healthy males (23.5+/-1.4 years) volunteered for the study. Surface electromyography (EMG) was used to evaluate muscle activities while the subjects walked in water, immersed to the level of the xiphoid process. HR responses were monitored continuously by a telemetry method. A "Flowmill" was used for this study, which involves a treadmill at the base of a water flume. Measurement of maximal voluntary contraction (MVC) of each tested muscle was undertaken prior to gait analysis. The %MVCs obtained from the paraspinal muscles, vastus medialis and tibialis anterior were all significantly greater when walking backward than when walking forward, for every experimental condition (P<0.05). HR responses tended to be greater while walking backward than when walking forward, with a statistical significance at fast speed (P<0.05). In conclusion, walking backward in water resulted in significantly greater muscle activation of the paraspinal muscles, vastus medialis and tibialis anterior compared with walking forward in water. These findings may be helpful in developing water-based exercise programs. PMID- 16713707 TI - The building blocks and motifs of RNA architecture. AB - RNA motifs can be defined broadly as recurrent structural elements containing multiple intramolecular RNA-RNA interactions, as observed in atomic-resolution RNA structures. They constitute the modular building blocks of RNA architecture, which is organized hierarchically. Recent work has focused on analyzing RNA backbone conformations to identify, define and search for new instances of recurrent motifs in X-ray structures. One current view asserts that recurrent RNA strand segments with characteristic backbone configurations qualify as independent motifs. Other considerations indicate that, to characterize modular motifs, one must take into account the larger structural context of such strand segments. This follows the biologically relevant motivation, which is to identify RNA structural characteristics that are subject to sequence constraints and that thus relate RNA architectures to sequences. PMID- 16713711 TI - Peroxisome proliferator-activated receptor delta (PPARdelta), a novel target site for drug discovery in metabolic syndrome. AB - The development of new treatments for metabolic syndrome is urgent project for decreasing the prevalence of coronary heart disease and diabetes mellitus in the advanced countries. Peroxisome proliferator-activated receptor (PPAR)alpha and gamma agonists have shed light on the treatment of hypertriglyceridemia and type 2 diabetes mellitus, respectively. Among PPARs, analysis of the PPARdelta functions is lagging behind because specific PPARdelta agonists have not been developed. The appearance of new PPARdelta agonists is brightening the prospects for elucidating the physiological role of PPARdelta. PPARdelta is a new target for the treatment of metabolic syndrome. In particular, the fact that fatty acid oxidation and energy dissipation in skeletal muscle and adipose tissue by PPARdelta agonists lead to improved lipid profile, reduced adiposity and insulin sensitivity is a breakthrough. It seems that treatment of PPARdelta agonists operate similarly to the caloric restriction and prolonged exercise. We suggest that the physiological role of PPARdelta may be an indicator for switching from glucose metabolism to fatty acid metabolism. To receive new benefits of PPARdelta agonists against metabolic syndrome by increasing fatty acid consumption in skeletal muscle and adipose tissue, we need to unveil more details on the functions of PPARdelta itself and its agonists in the future. PMID- 16713713 TI - Studies on ternary metallo-beta lactamase-inhibitor complexes using electrospray ionization mass spectrometry. AB - Metallo-beta-lactamases (MBLs) are targets for medicinal chemistry as they mediate bacterial resistance to beta-lactam antibiotics. Electrospray-ionization mass spectrometry (ESI-MS) was used to study the inhibition by a set of mercaptocarboxylates of two representative MBLs with different optimal metal stoichiometries for catalysis. BcII is a dizinc MBL (Class B1), whilst the CphA MBL (Class B2) exhibits highest activity with a single zinc ion in the active site. Experimental parameters for the detection of the metallo-enzyme and the metallo-enzyme-inhibitor complexes were evaluated and optimized. Following investigations on the stoichiometry of metal binding, the affinity of the inhibitors was investigated by measuring the relative abundance of the complex compared to the metalloprotein. The results for the BcII enzyme were in general agreement with solution assays and demonstrated that the inhibitors bind to the dizinc form of the BcII enzyme. The results for the CphA(ZnII) complex unexpectedly revealed an increased affinity for the binding of a second metal ion in the presence of thiomandelic acid. The results demonstrate that direct ESI-MS analysis of enzyme:inhibitor complexes is a viable method for screening inhibitors and for the rapid assay of the enzyme:metal:inhibitor ratios. PMID- 16713714 TI - Autoimmunity in systemic lupus erythematosus: integrating genes and biology. AB - Susceptibility to the autoimmune phenotype of systemic lupus erythematosus (SLE) is heritable. Linkage analysis and recent advances in the field of single nucleotide polymorphisms (SNPs) have resulted in the identification of several genetic loci and functional allelic variants of signaling proteins which have become the mainstay of understanding disease susceptibility and exploring the basis of autoimmunity in SLE. However, genetic heterogeneity and possible epistatic interactions among genetic elements have precluded replication of these findings in multiple population groups and thus complicated their interpretation. In this regard, the discovery that a plethora of normal signaling proteins are expressed in abnormal amounts in immune cells from patients with SLE has gained significance. Thus, the key to precise elucidation of the pathologic basis of autoimmunity in SLE lies in tying genetics and disease biology. This review highlights recent discoveries of important functional genetic variants and altered expression of normal signaling proteins that network together to disrupt peripheral tolerance and initiate the autoimmune process in SLE. PMID- 16713712 TI - Determination of peptide and protein ion charge states by Fourier transformation of isotope-resolved mass spectra. AB - We report an automated method for determining charge states from high-resolution mass spectra. Fourier transforms of isotope packets from high-resolution mass spectra are compared to Fourier transforms of modeled isotopic peak packets for a range of charge states. The charge state for the experimental ion packet is determined by the model isotope packet that yields the best match in the comparison of the Fourier transforms. This strategy is demonstrated for determining peptide ion charge states from "zoom scan" data from a linear quadrupole ion trap mass spectrometer, enabling the subsequent automated identification of singly- through quadruply-charged peptide ions, while reducing the numbers of conflicting identifications from ambiguous charge state assignments. We also apply this technique to determine the charges of intact protein ions from LC-FTICR data, demonstrating that it is more sensitive under these experimental conditions than two existing algorithms. The strategy outlined in this paper should be generally applicable to mass spectra obtained from any instrument capable of isotopic resolution. PMID- 16713715 TI - Spontaneous development of autoimmune arthritis due to genetic anomaly of T cell signal transduction: Part 1. AB - A point mutation of the gene encoding ZAP-70, a key signal transduction molecule in T cells, results in spontaneous development of T cell-mediated autoimmune arthritis in mice homozygous for the mutation. The genetic anomaly alters differentiation and selection of T cells in the thymus, leading to thymic production of arthritogenic autoimmune T cells. The arthritogenic T cells persist in the periphery and elicit arthritis when activated by microbial agents that stimulate innate immunity. This model is instrumental in understanding how genetic variations in T cell signal transduction, together with environmental influences, contribute to the development of autoimmune disease. PMID- 16713716 TI - Intravascular effect in velocity-selective arterial spin labeling: the choice of inflow time and cutoff velocity. AB - Velocity-selective arterial spin labeling (VS-ASL) tags spins on a basis of flow velocity, instead of spatial distribution that has been commonly adopted in conventional ASL techniques. VS-ASL can potentially generate tags that are very close to the imaging plane and whereby avoid the error source of transit delay (deltat) variation independent of inflow time (TI). In practice, however, TI of VS-ASL should still be chosen with caution with respect to intravascular signal and cutoff velocity (V(c)). The presented study takes advantage of multiple TI and V(c) to systematically investigate the intravascular effect. Results demonstrate the presence of significant signal from large vessels in VS-ASL images for V(c) down to 4 cm/s. For perfusion measurement in human brain, low V(c) (<4 cm/s) is recommended. With V(c) = 2 cm/s, quantitative cerebral blood flow is 72.8 ml/100 ml/min, which is in agreement with the reported range using conventional ASL methods. In field strength of 3 T, numerical simulation shows that optimal signal-to-noise ratio efficiency can be achieved with TR/TI = 2092 ms/1664 ms for single slice and 4493 ms/1404 ms for slab imaging. PMID- 16713717 TI - Spatial flow-volume dissociation of the cerebral microcirculatory response to mild hypercapnia. AB - The spatial and temporal response of the cerebral microcirculation to mild hypercapnia was investigated via two-photon laser-scanning microscopy. Cortical vessels, traversing the top 200 microm of somatosensory cortex, were visualized in alpha-chloralose-anesthetized Sprague-Dawley rats equipped with a cranial window. Intraluminal vessel diameters, transit times of fluorescent dextrans and red blood cells (RBC) velocities in individual capillaries were measured under normocapnic (PaCO2= 32.6 +/- 2.6 mm Hg) and slightly hypercapnic (PaCO2= 45 +/- 7 mm Hg) conditions. This gentle increase in PaCO2 was sufficient to produce robust and significant increases in both arterial and venous vessel diameters, concomitant to decreases in transit times of a bolus of dye from artery to venule (14%, P < 0.05) and from artery to vein (27%, P < 0.05). On the whole, capillaries exhibited a significant increase in diameter (16 +/- 33%, P < 0.001, n = 393) and a substantial increase in RBC velocities (75 +/- 114%, P < 0.001, n = 46) with hypercapnia. However, the response of the cerebral microvasculature to modest increases in PaCO2 was spatially heterogeneous. The maximal relative dilatation (range: 5-77%; mean +/- SD: 25 +/- 34%, P < 0.001, n = 271) occurred in the smallest capillaries (1.6 microm-4.0 microm resting diameter), while medium and larger capillaries (4.4 microm-6.8 microm resting diameter) showed no significant changes in diameter (P > 0.08, n = 122). In contrast, on average, RBC velocities increased less in the smaller capillaries (39 +/- 5%, P < 0.002, n = 22) than in the medium and larger capillaries (107 +/- 142%, P < 0.003, n = 24). Thus, the changes in capillary RBC velocities were spatially distinct from the observed volumetric changes and occurred to homogenize cerebral blood flow along capillaries of all diameters. PMID- 16713718 TI - Glucagon challenge in the rat: a robust method for the in vivo assessment of Glycogen phosphorlyase inhibitor efficacy. AB - INTRODUCTION: Glycogen phosphorlyase inhibitors (GPi) act on the glycogenolytic pathway decreasing hepatic glucose output, making them potential candidates for Type 2 diabetes treatment. We established a robust in vivo method to assess GPis efficacy utilising glucagon-stimulated glycogenolysis. METHODS: Blood glucose was monitored in both male AP Wistar and AP Zucker rats using tail prick samples pre- and post intraperitoneal or subcutaneous glucagon administration. The effect of glycogen phosphorylase inhibitors GPi296 (6-60 mg kg(-1) po) and DAB (5 mg kg(-1) po) upon glucose response to subcutaneous glucagon were examined in both strains. RESULTS: In the Wistar rat glucagon induced dose related increases in blood glucose, with the maximum increase occurring 20 min post dose (4.0+/-0.88 mmol l( 1), intraperitoneal; and 2.8+/-0.72 mmol l(-1), subcutaneous, ns). Intraperitoneal glucagon administration produced shorter duration blood glucose elevation than observed with the subcutaneous route of administration. In the Zucker rat, no differences were observed between the 10 and 13 week old rats in response to glucagon (3-200 microg kg(-1) subcutaneous). The maximum blood glucose increase was lower in the Wistar rat compared to the Zucker rats (2.9+/ 0.20 vs 7.7+/-1.22 mmol l(-1), P<0.0000018). GPi296 and DAB both produced similar inhibition in each strain. DISCUSSION: Subcutaneous glucagon administration induced more sustained increases in blood glucose than intraperitoneal administration. Blood glucose response to glucagon was higher in the Zucker rat compared to the Wistar rat; there was no difference in inhibition mediated by either GPi296 or DAB between the two strains. We believe that subcutaneous glucagon administration produces a robust model for the assessment of GPis in either rat strain. PMID- 16713719 TI - MR measurement of articular cartilage thickness distribution in the hip. AB - OBJECTIVE: To develop a method to determine the distribution of articular cartilage in the hip and to evaluate the potential of the method in a study of normal weight-bearing effects in asymptomatic young volunteers. DESIGN: Six volunteers were scanned after periods of standing and lying supine, using 3D gradient-echo magnetic resonance imaging (MRI). The protocol was repeated for two successive weeks to determine reproducibility. The femoral and acetabular cartilage layers were segmented as a single unit and thickness distribution maps were calculated using a spherical bone model as a frame of reference. Thickness maps were combined over the population using the bone model and post-weight bearing and post-resting maps were compared. RESULTS: Mean thickness values were compared using an analysis of variance and a significant increase in cartilage thickness of 0.05 mm (P=0.02) was observed. The reproducibility of the method, assessed using test-retest coefficient of variation was 2.5%. CONCLUSIONS: The technique is reproducible, sensitive to sub-millimetre changes in thickness and may be useful in monitoring changes due to disease progression in patients with arthritis of the hip. PMID- 16713720 TI - Magnetic resonance imaging (MRI) of articular cartilage in knee osteoarthritis (OA): morphological assessment. AB - OBJECTIVE: Magnetic resonance imaging (MRI) is a three-dimensional imaging technique with unparalleled ability to evaluate articular cartilage. This report reviews the current status of morphological assessment of cartilage with quantitative MRI (qMRI), and its relevance for identifying disease status, and monitoring progression and treatment response in knee osteoarthritis (OA). METHOD: An international panel of experts in MRI of knee OA, with direct experience in the analysis of cartilage morphology with qMRI, reviewed the existing published and unpublished data on the subject, and debated the findings at the OMERACT-OARSI Workshop on Imaging technologies (December 2002, Bethesda, MA) with scientists and clinicians from academia, the pharmaceutical industry and the regulatory agencies. This report reviews (1) MRI pulse sequence considerations for morphological analysis of articular cartilage; (2) techniques for segmenting cartilage; (3) semi-quantitative scoring of cartilage status; and (4) technical validity (accuracy), precision (reproducibility) and sensitivity to change of quantitative measures of cartilage morphology. RESULTS: Semi quantitative scores of cartilage status have been shown to display adequate reliability, specificity and sensitivity, and to detect lesion progression at reasonable observation periods (1-2 years). Quantitative assessment of cartilage morphology (qMRI), with fat-suppressed gradient echo sequences, and appropriate image analysis techniques, displays high accuracy and adequate precision (e.g., root-mean-square standard deviation medial tibia=61 microl) for cross-sectional and longitudinal studies in OA patients. Longitudinal studies suggest that changes of cartilage volume of the order of -4% to -6% occur per annum in OA in most knee compartments (e.g., -90 microl in medial tibia). Annual changes in cartilage volume exceed the precision errors and appear to be associated with clinical symptoms as well as with time to knee arthroplasty. CONCLUSIONS: MRI provides reliable and quantitative data on cartilage status throughout most compartments of the knee, with robust acquisition protocols for multi-center trials now being available. MRI of cartilage has tremendous potential for large scale epidemiological studies of OA progression, and for clinical trials of treatment response to structure modifying OA drugs. PMID- 16713721 TI - Aggrecan core protein of a certain length is protective against hand osteoarthritis. AB - OBJECTIVE: To study the contribution of aggrecan VNTR (variable number of tandem repeats) polymorphism to clinically differing manifestations of hand osteoarthritis (OA). DESIGN: Five hundred thirty Finnish females representing two academically similar occupations with completely diverse exposure to hand load were included. Radiographs of hands were analysed, the OA findings were graded and the subjects were divided into categories. Aggrecan VNTR alleles were identified by Southern hybridization. Statistical analyses were used to compare joint involvement and pathological findings with the prevalences of the alleles and genotypes. RESULTS: Subjects homozygous for the most common aggrecan VNTR allele, A27 with 27 repeats, had a significantly lower risk of hand OA, with OR 0.46 (95% CI 0.27-0.78) for OA of grade 2 or more. Our results suggest that carrying two copies of the alleles with less than 27 repeats could predispose a subject to a severe hand OA (OR 2.45, 95% CI 1.17-5.12) and carrying two copies of the alleles with more than 27 repeats also increases the risk of the disease (OR 1.73, 95% CI 1.03-2.89). CONCLUSIONS: These findings indicate that allele A27 provides protection from hand OA and that alleles shorter or longer than this may predispose subjects to the disease. Furthermore, they suggest that a certain number of tandem repeats provide for optimal functioning of the aggrecan molecule and that the contribution of genetic factors to the development of hand OA may be even more important than that of environmental factors. PMID- 16713722 TI - MRI and non-cartilaginous structures in knee osteoarthritis. AB - Magnetic resonance imaging (MRI) provides a sensitive tool for examining all the structures involved in the osteoarthritis (OA) process. While much of the MRI literature previously focussed on cartilage, there is increasing research on whole-organ evaluation and including features such as synovitis, bone marrow edema, and meniscal and ligamentous pathology. The aim of this session at the Outcome Measures in Rheumatology Clinical Trials (OMERACT)-Osteoarthritis Research Society International (OARSI) Workshop for Consensus in Osteoarthritis Imaging was to describe the current MRI methods for identifying and quantifying non-cartilaginous structures and review their associations with both OA symptoms and structural progression. Although there is much experience in measuring synovitis (derived from the rheumatoid arthritis literature), only one study has reported an association of MRI-detected synovitis and effusions with OA pain. Bone marrow edema lesions, which may represent areas of trabecular remodelling, have been associated with pain and compartment-specific structural deterioration. MRI studies have confirmed the frequency and importance of meniscal damage in progressive cartilage loss, but not related such damage to symptoms. Osteophytes have been associated with cartilage loss and malalignment to the side of the osteophyte. Ligament damage, including anterior cruciate ligament tears, has been found more commonly than expected in painful OA knees. Improvements in quantitative and semi-quantitative assessments of non-cartilage features will greatly assist understanding of the OA process and its response to therapy. PMID- 16713723 TI - Quantitative structure and retention relationships for gas chromatographic data: application to alkyl pyridines on apolar and polar phases. AB - Quantitative structure and retention relationships (QSRR) have been developed to model gas chromatographic retention data of alkyl pyridines on apolar (branched alkane) and polar (primary alcohol) stationary phases. The retention properties analyzed are Kovats retention index, I; partial molar enthalpy, DeltaH; partial molar entropy, DeltaS and partition coefficient, log K. Using the seven valence molecular connectivity indices (chi) calculated for the 18 alkyl pyridines, regression models are generated to predict the retention properties. The best model (model A) obtained with the descriptors ((1)chi(P)(V), (3)chi(P)(V) and (6)chi(CH)(V) was unable to produce a satisfactory statistical performance and correct order of elution. The model has been modified (model B) by including steric parameter s, which has been empirically derived by considering the steric effects due to the presence of alkyl groups at the ortho and meta positions. The modified model predicts the correct order of elution for all the alkyl pyridines and good correlation coefficients, r. The r values obtained for I, DeltaH, DeltaS and log K are: r=0.955, 0.975, 0.984 and 0.955 (model A) become r=0.999, 0.996, 0.990 and 0.999 (model B) on the apolar stationary phase and r=0.931, 0.926, 0.911 and 0.931 (model A) become r=0.999, 0.992, 0.958 and 0.999 (model B) on the polar stationary phase. PMID- 16713724 TI - The genetics of Salmonella genomic island 1. AB - Multidrug-resistant Salmonella enterica serovar Typhimurium phage type DT104, resistant to ampicillin, chloramphenicol/florfenicol, streptomycin, sulfonamides, and tetracycline, has disseminated worldwide. The resistance genes reside on the 43-kb Salmonella genomic island 1 (SGI1), which is transferable. Drug-resistant variants of SGI1 have been identified in numerous serotypes. Strains harboring SGI1 may be more virulent and have a tendency to rapidly disseminate. PMID- 16713725 TI - Evolution of the virulence plasmids of non-typhoid Salmonella and its association with antimicrobial resistance. AB - Among more than 2,500 serovars, eight contain a virulence plasmid, including medically important Salmonella enterica serovars Choleraesuis, Dublin, Enteritidis, and Typhimurium. These serovar-specific virulence plasmids vary in size, but all contain the spv operon, which plays a role in the expression of the virulence. Genetically, these virulence plasmids are likely derived from a common ancestral plasmid possessing virulence-related genes and loci. Based on the analysis of the available DNA sequences of the plasmids, the phylogenetic path may be split into two: pSPV (virulence plasmid of S. Gallinarum-Pullorum) acquires an incompatibility-related locus that differs from that of the others. At some point, pSCV (virulence plasmid of S. Choleraesuis) and pSDV (virulence plasmid of S. Dublin) lose oriT by recombination or simply by deletion, making the two unable to be mobilized. On the other hand, pSEV (virulence plasmid of S. Enteritidis) also loses some DNA by deletion but not as extensively as pSCV, and therefore pSEV is closest to pSTV (virulence plasmid of S. Typhimurium) both genetically and biologically. The pSTV shows the least alternation during the evolution. There are two types of pSDV. pSDVu recombines with non-virulence 36.6 kb plasmid to acquire additional incompatibility trait to form pSDVr. Recent reports indicated that S. Choleraesuis and S. Typhimurium could generate different types of hybrid plasmids, which consisted of the serovar-specific virulence plasmid and an array of resistance gene cassettes. The recombination gives Salmonella a survival advantage in an unfavorable drug environment. The integration of resistance genes and additional replicons into a Salmonella virulence plasmid constitutes a new and interesting example of plasmid evolution and poses a serious threat to public health. PMID- 16713726 TI - Mechanisms of fluoroquinolone and macrolide resistance in Campylobacter spp. AB - The incidence of human Campylobacter infections is increasing worldwide, as well as the proportion of isolates resistant to fluoroquinolones and/or macrolides, the drugs of choice to treat campylobacteriosis. In this review, we report recent developments in the understanding of the resistance mechanisms to fluoroquinolones and macrolides. In addition, we will discuss the recent findings on multidrug resistance in Campylobacter spp. PMID- 16713728 TI - Kinetics and mechanism of nickel ferrite formation under high temperature ultrasonic treatment. AB - The effect of simultaneous ultrasonic and thermal treatment on kinetics and mechanism of nickel ferrite formation was studied. It was established that sonication leads to notable increase of the mean rate of this reaction and decrease of effective activation energy from 190+/-5 to 125+/-7 kJ/mol. XRD data show that ultrasonic treatment significantly affects the microstructure of both initial reagent (Fe(2)O(3)) and reaction product (NiFe(2)O(4)) thus promoting formation of well developed reaction zone. A general model of ferrite formation mechanism under high temperature ultrasonic treatment was proposed. PMID- 16713727 TI - Repeated respiratory Mycoplasma pneumoniae infections in mice: effect of host genetic background. AB - Respiratory Mycoplasma pneumoniae (Mp) infection is involved in several acute and chronic lung diseases including community-acquired pneumonia, asthma and chronic obstructive pulmonary disease. In the chronic disease process, recurrent respiratory bacterial infections could occur, which may result in varying degrees of symptoms and lung inflammation among patients. However, the lung immunologic differences of host responses to repeated bacterial (i.e., Mp) infections remain to be determined. In the present study, we examined cellular and humoral responses to multiple (up to 3) Mp infections in two genetically different strains of mice (BALB/c and C57BL/6). Mice were intranasally inoculated with one Mp infection, two or three Mp infections (4 weeks apart), and sacrificed on days 3, 7 and 14 after the last Mp infection. Overall, compared to C57BL/6 mice, BALB/c mice demonstrated a significantly higher degree of lung tissue inflammatory cell infiltrate, BAL cellularity, and release of pro-inflammatory cytokines (TNF-alpha, keratinocyte-derived chemokine (KC, a mouse homolog of human chemokine Gro-alpha [CXCL1], and IFN-gamma). In addition, BALB/c mice presented higher levels of serum Mp-specific IgG and IgM, but not IgA. Consistently with lung and serum data, Mp load in BAL and lung specimens was significantly higher in BALB/c mice than C57BL/6 mice. Moreover, repeated Mp infections in BALB/c, but not C57BL/6 mice, produced a greater inflammatory response than did a single Mp infection. Our results suggest that hosts with different genetic background may have different susceptibility to repeated respiratory Mp infections along with inflammatory responses. PMID- 16713729 TI - Resistance proteins: molecular switches of plant defence. AB - Specificity of the plant innate immune system is often conferred by resistance (R) proteins. Most R proteins contain leucine-rich repeats (LRRs), a central nucleotide-binding site (NBS) and a variable amino-terminal domain. The LRRs are mainly involved in recognition, whereas the amino-terminal domain determines signalling specificity. The NBS forms part of a nucleotide binding (NB)-ARC domain that presumably functions as a molecular switch. The conserved nature of NB-ARC proteins makes it possible to map mutations of R protein residues onto the crystal structures of related NB-ARC proteins, providing hypotheses for the functional roles of these residues. A functional model emerges in which the LRRs control the molecular state of the NB-ARC domain. Pathogen recognition triggers nucleotide-dependent conformational changes that might induce oligomerisation, thereby providing a scaffold for activation of downstream signalling components. PMID- 16713730 TI - Type III effector proteins: doppelgangers of bacterial virulence. AB - Bacterial pathogens have co-evolved with their hosts in their ongoing quest for advantage in the resulting interaction. These intimate associations have resulted in remarkable adaptations of prokaryotic virulence proteins and their eukaryotic molecular targets. An important strategy used by microbial pathogens of animals to manipulate host cellular functions is structural mimicry of eukaryotic proteins. Recent evidence demonstrates that plant pathogens also use structural mimicry of host factors as a virulence strategy. Nearly all virulence proteins from phytopathogenic bacteria have eluded functional annotation on the basis of primary amino-acid sequence. Recent efforts to determine their three-dimensional structures are, however, revealing important clues about the mechanisms of bacterial virulence in plants. PMID- 16713731 TI - Autophagy in the control of programmed cell death. AB - Programmed cell death (PCD) is essential for plant development and immunity. Localized PCD is associated with the hypersensitive response (HR), which is a constituent of a successful plant innate immune response. Plants have developed mechanisms to meticulously prevent HR-PCD lesions from spreading. Our understanding of these mechanisms is still in its incipient stages. A recent study demonstrated that autophagy, a universally conserved process of macromolecule turnover, plays a pivotal role in controlling HR-PCD. The molecular identity of the mediators between the PCD and HR pathways is still obscure, but recent work has begun to shed light on the relationship between HR-PCD and autophagy and to suggest possible mechanisms for the regulation of these pathways. PMID- 16713732 TI - Mutualism and parasitism: the yin and yang of plant symbioses. AB - Plants are solar-powered sugar factories that feed a multitude of other organisms. Many of these organisms associate directly with host plants to gain access to the plant's photosynthates. Such symbioses encompass a wide collection of styles ranging from mutualistic to commensal and parasitic. Among these, the mutualistic arbuscular mycorrhizal (AM) symbiosis is one of the evolutionarily oldest symbioses of plants, relying on the formation of an intimate relationship between fungi of the Glomeromycota and roots of the majority of vascular flowering plants. In this symbiosis, the fungus intracellularly colonizes living root cells, implying the existence of an extreme form of compatibility. Interestingly, molecular events that happen in the plant in response to mycorrhizal colonization also occur in other beneficial and, as recently shown, even antagonistic plant symbioses. Thus, basic 'compatibility modules' appear to be partially conserved between mutualism and parasitism. PMID- 16713733 TI - Secondary metabolite toxins and nutrition of plant pathogenic fungi. AB - Fungal pathogens derive nutrition from the plants they invade. Some fungi can subvert plant defence responses such as programmed cell death to provide nutrition for their growth and colonisation. Secondary metabolite toxins produced by fungi often play a role in triggering these responses. Knowledge of the biosynthesis of these toxins, and the availability of fungal genome sequences and gene disruption techniques, allows the development of tools for experiments aimed at discovering the role of such toxins in triggering plant cell death and plant disease. PMID- 16713734 TI - Involvement of the drug efflux protein TolC in mutagenicity induced by MNNG or Trp-P-2. AB - In the development of mutation assay systems, a number of approaches have been performed with a particular view to improve sensitivity. The inhibition of mutagen-efflux from tester bacteria might lead to increased mutagenic activity as the concentration of mutagen increases inside the cell. In this study, we constructed a series of Escherichia coli CC strains lacking the TolC protein to determine if mutation is actually enhanced by the inhibition of mutagen reflux. TolC is an outer-membrane protein that forms part of an excretion system in E. coli. The frequency of induction of mutations by N-methyl-N'-nitro-N nitrosoguanidine (MNNG), N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG) and ethyl methanesulfonate (EMS) were significantly higher in TolC-deficient strain KA796 1/CC102 than in TolC-proficient strains, especially that of MNNG was seven times higher and detected at lower doses than in the parent strain. In a KA796-1/CC108 TolC-deficient strain, mutation induced by Trp-P-2 was detected at significant levels, even at low doses that did not induce detectable levels of mutation in the parent strain KA796/CC108. When the wild-type E. coli tolC gene was introduced into a strain lacking the gene, TolC function was restored and the frequency of induction by MNNG became similar to that of the wild-type. In contrast, introduction of a mutant tolC gene did not complement the TolC deficiency and the frequency of MNNG-induced mutations remained high. These results suggest that some mutagens are excreted at least in part via the TolC system, and that the lack of functional TolC increases the susceptibility of bacteria to many mutagens. PMID- 16713735 TI - A high-throughput method to measure the sensitivity of yeast cells to genotoxic agents in liquid cultures. AB - The sensitivity of yeast Saccharomyces cerevisiae to DNA damaging agents is better represented when cells are grown in liquid media than on solid plates. However, systematic assessment of several strains that are grown in different conditions is a cumbersome undertaking. We report an assay to determine cell growth based on automatic measurements of optical densities of very small (100 microl) liquid cell cultures. Furthermore, an algorithm was elaborated to analyze large data files obtained from the cell growth curves, which are described by the growth rate--that starts at zero and accelerates to the maximal rate (mu(m))--and by the lag time (lambda). Cell dilution spot test for colony formation on solid media and the growth curve assay were used in parallel to analyze the phenotypes of cells after treatments with three different classes of DNA damaging agents (methyl methanesulfonate, bleomycin, and ultraviolet light). In these experiments the survival of the WT (wild type) and a number of DNA repair-deficient strains were compared. The results show that only the cell growth curve assay could uncover subtle phenotypes when WT cells, or mutant strains that are only weakly affected in DNA repair proficiency, were treated with low doses of cytotoxic compounds. The growth curve assay was also applied to establish whether histone acetyltransferases and deacetylases affect the resistance of yeast cells to UV irradiation. Out of 20 strains tested the sir2delta and rpd3delta cells were found to be more resistant than the WT, while gcn5delta and spt10delta cells were found to be more sensitive. This new protocol is sensitive, provides quantifiable data, offers increased screening capability and speed compared to the colony formation test. PMID- 16713736 TI - Familial occurrence of isolated non-compaction cardiomyopathy. AB - BACKGROUND AND AIMS: Isolated left ventricular non-compaction cardiomyopathy (LVNC) may have an autosomal dominant or X-linked recessive inheritance. We focus on the familial occurrence of LVNC after misdiagnosing this disorder in symptomatic patients in two families. After identification of the index patient we studied the families more intensively in order to unmask affected family members. METHODS AND RESULTS: LVNC was defined as an end-systolic non-compacted subendocardial layer of the left ventricular wall of at least twice the thickness of the subepicardial compacted layer (2D echocardiogram and MRI). This was studied in 13 patients in 2 families (A and B). LVNC was found in 3 out of 11 patients in family A. The grandmother was asymptomatic. Her daughter suffered from recurrent syncope and heart failure. Her daughter received a cardiac transplant because of progressive heart failure at the age of 14years. In family B, LVNC was found in 2 patients, a father and his son and presumed in a brother and a sister of the father who died suddenly at the age of 17 and 21years, respectively. CONCLUSIONS: In all symptomatic patients, proven LVNC was previously misdiagnosed as hypertrophic or dilated cardiomyopathy. Misdiagnosis may lead to insufficient treatment and will misdirect targeted molecular genetic analysis. LVNC was identified in seven patients in two families. Family screening may unmask affected family members for primary prevention including anti coagulation and ICD-therapy. PMID- 16713737 TI - Long-term effects of levosimendan infusion on inflammatory processes and sFas in patients with severe heart failure. AB - BACKGROUND: The calcium sensitizer levosimendan improves myocardial contractility in patients with heart failure, although its effects on inflammation and apoptosis are unknown. AIM: To examine the effects of levosimendan on markers of inflammation and apoptosis, over a period of 30 d following a 24 h infusion, in patients with heart failure. METHODS: Thirty four patients with severe heart failure were randomised to receive a 24 h infusion of levosimendan or placebo, in a double-blind trial. Haemodynamic evaluation and blood sampling were performed at baseline, 24 h, 30 h, 48 h, 7 d and 30 d after the end of the infusion. RESULTS: Seven patients (1 levosimendan, 6 placebo), were excluded during follow up. In the remaining 27 patients, levosimendan decreased serum IL-6 and sFAS, 24 h after the infusion (p<0.01 and p<0.05 vs baseline), an effect sustained for 7 30 d. Serum TNF-alpha and sTNF-R1 were decreased between 48 h (p<0.01 vs baseline for both) and 7 d (p<0.05 vs baseline for sTNF-R1) after infusion. Serum sTNF-R2 was decreased at 24 h (p<0.05 vs baseline) and remained lower than baseline for at least 7 d (p<0.05). CONCLUSIONS: These findings indicate that levosimendan decreases the expression of proinflammatory cytokines, TNF-alpha receptors and sFAS, immediately after infusion, an effect which persists for 7-30 d. PMID- 16713738 TI - A blueprint for success: integration of neglected tropical disease control programmes. AB - The rapid expansion of chemotherapy-based control programmes for neglected tropical diseases has been catalysed by funding from the Bill and Melinda Gates Foundation, donations of several drugs from pharmaceutical manufacturers, and the reduced price of the drug praziquantel. Focussing on lymphatic filariasis, schistosomiasis and soil-transmitted helminthiasis, we review here the progress made to date with the implementation and integration of large-scale control programmes. Unresolved issues include a means for rapid identification of communities at highest risk of co-morbidity, cost-effective approaches for integrating the technical interventions into setting-specific packages, and determination of the most appropriate and sustainable delivery systems. PMID- 16713739 TI - The IPTi Consortium: research for policy and action. AB - The results of a randomized controlled trial in Tanzania suggest that intermittent preventive treatment in infants (IPTi), delivered through the Expanded Program on Immunization, might be a useful approach to controlling malaria in countries where it is endemic. An international consortium of research collaborations, involving the World Health Organization and United Nation's Children's Fund, is now evaluating IPTi in a range of different settings to generate robust and compelling evidence to guide policy. This review summarizes the available information on IPTi and presents the consortium's approach to determining whether IPTi might be a valuable additional strategy in programs to control malaria. PMID- 16713740 TI - Progress towards eliminating lymphatic filariasis in Zanzibar: a model programme. AB - Programmes to eliminate lymphatic filariasis are underway in ten countries of sub Saharan Africa, and in several programmes outside Africa five rounds of mass drug administration (MDA) are being completed. In Africa, Egypt and Zanzibar have completed five rounds of MDA. Zanzibar was the first country to complete five rounds of treatment using a combination of albendazole and ivermectin, reducing both the prevalence and intensity of Wuchereria bancrofti. Characteristics of the Zanzibar programme serve as a model for other countries: factors crucial to its success include high-level political commitment, the development of appropriate social mobilization strategies, the involvement of communities in drug distribution, and the introduction of morbidity management for individuals with lymphoedema. PMID- 16713741 TI - STAT4 deficiency reduces autoantibody production and glomerulonephritis in a mouse model of lupus. AB - To determine the respective role of the IL-12 and IL-4 pathways in the pathogenesis of systemic lupus erythematosus, we bred the Stat4 and Stat6 null alleles onto the lupus-prone mouse B6.TC, which is a congenic derivative of NZM2410. This model is characterized by abnormal splenocyte expansion, distribution and architecture, T cell activation, peripheral B cell development, production of anti-nuclear antibodies, and proliferative glomerulonephritis. STAT4 deficiency normalized the expression of each of these disease markers toward or to C57BL/6 levels. In contrast, STAT6 deficiency impacted splenocyte expansion and architecture, T cell activation, and anti-nuclear autoantibody production, but without any significant effect on B cell development or renal pathology. These results show that the IL-12/STAT4 pathway is involved in multiple disease-associated phenotypes in the B6.TC mouse. In contrast, the IL 4/STAT6 pathway regulates only a subset of disease markers that did not affect renal pathology. PMID- 16713742 TI - Vaccination of mice with replication-defective human immunodeficiency virus induces cellular and humoral immunity and protects against vaccinia virus-gag challenge. AB - Here we describe as a potential vaccine candidate a replication-defective HIV that encodes multiple viral genes in addition to a cassette that includes both truncated cyclin T1 and an autofluorescent protein. After confirming functionality of the cyclin T1, we immunized mice intramuscularly once or twice with the replication-defective HIV vector pseudotyped with vesicular stomatitis virus (VSV) G protein (RD HIV), a plasmid encoding CMV-driven gag (gag DNA), or adenovirus gag (Ad5-gag). Capsid-specific antibody titers following RD HIV immunization were >10(6)/ml and approximately equivalent to those induced by gag DNA and Ad5-gag. Antibodies against the autofluorescent protein and VSV G were also detected. After RD HIV immunization ELISpot assays demonstrated Gag-specific interferon-gamma (IFN-gamma) SFU equivalent to that of Ad5-gag and fourfold greater than that of gag DNA. HIV polymerase-specific IFN-gamma SFU values were similar, and boosting increased both antibody titers and the IFN-gamma response. Challenge using vaccinia virus (VV)-gag demonstrated significantly lower recoverable VV for RD HIV-immunized mice compared to controls. No significant differences were observed in vaccinated mice challenged with wild-type VV. This study demonstrates the efficacy of RD HIV in conferring HIV-specific immunity and protection in mice and suggests its potential use in humans as either a prophylactic or a therapeutic vaccine. PMID- 16713744 TI - Endoscopic ultrasonography and prognosis of esophageal cancer. AB - BACKGROUND & AIMS: Endoscopic ultrasonography (EUS) staging is used for management of esophageal cancer, but its effect on the outcome of patients is unknown. Our aim was to study the association of receipt of EUS and overall survival in a cohort of patients with esophageal cancer. METHODS: All persons 65 years or older who were diagnosed with esophageal cancer between January 1994 and December 1999 in the Surveillance, Epidemiology, and End Results (SEER)-Medicare linked database were identified. Relevant demographic, cancer-specific information and EUS procedural information were extracted. RESULTS: Two thousand eight hundred thirty patients with esophageal cancer (48% squamous cell cancer) were eligible for analysis. Only 303 (10.7%) patients underwent a EUS examination. Patients who had EUS evaluation were more likely to undergo esophageal resection (21.1% vs 14.7%, P = .01) and more likely to have received adjuvant therapy (11.2% vs 6.7%, P = .008). When adjusted for age at diagnosis, race, gender, comorbidity, histology, and tumor stage, receipt of EUS was associated with a reduced risk of death (relative hazard, 0.594; 95% confidence interval, 0.52-0.68; P = .001). CONCLUSIONS: Undergoing EUS in patients with esophageal cancer is independently associated with improved survival, possibly because of improved stage-appropriate management such as use of adjuvant therapy and surgical resection. PMID- 16713745 TI - Do cytokine concentrations in pancreatic juice predict the presence of pancreatic diseases? AB - BACKGROUND & AIMS: Cytokine concentration in pancreatic juice of patients with pancreatic disease is unknown. Secretin stimulation allows endoscopic collection of pancreatic juice secreted into the duodenum. We aimed to evaluate the cytokine concentrations in pancreatic juice of patients with abdominal pain to discriminate presence from absence of pancreatic disease. METHODS: From January 2003-December 2004, consecutive patients with abdominal pain compatible with pancreatic origin were enrolled. Patients underwent upper endoscopy. Intravenous secretin (0.2 mug/kg) was given immediately before scope intubation. Pancreatic juice collected from the duodenum was immediately snap-frozen in liquid nitrogen until assays were performed. Pancreatic juice levels of interleukin-8, interleukin-6, intercellular adhesion molecule 1, and transforming growth factor beta 1 were measured by modified enzyme-linked immunosorbent assays. The final diagnosis was made by the primary gastroenterologist on the basis of medical history; laboratory, endoscopic, and imaging studies; and clinical follow-up. Fisher exact test and Kruskal-Wallis rank sum test were used for statistical analysis. RESULTS: Of 130 patients screened, 118 met the inclusion criteria. Multivariate analysis revealed that only interleukin-8 was able to discriminate between normal pancreas and chronic pancreatitis (P = .011), pancreatic cancer (P = .044), and the presence of pancreatic diseases (P = .007). Individual cytokine concentrations were not significantly different in chronic pancreatitis compared with pancreatic cancer. CONCLUSIONS: Cytokine levels can be measured in pancreatic juice obtained from the duodenum without direct cannulation of the pancreatic duct. Interleukin-8 concentration in pancreatic juice can be used to discriminate between normal pancreas and patients with pancreatic disease. This is a relatively simple and noninvasive method to aid in the diagnosis of pancreatic diseases. PMID- 16713746 TI - Endoscopic submucosal dissection of esophageal squamous cell neoplasms. AB - BACKGROUND & AIMS: Endoscopic submucosal dissection (ESD) has recently been developed for en bloc resection of stomach neoplasms, which results in high tumor eradication rates as well as a modality for the precise histologic assessment of the entire lesion. Application of the technique is desirable for esophageal squamous cell neoplasms (SCNs), but there have been no reports on the use of this procedure in the esophagus. METHODS: An ESD with methods similar to those used for resections of early gastric cancer was performed on 58 consecutive esophageal SCNs with preoperative diagnoses of intraepithelial neoplasm or intramucosal invasive carcinoma occurring in 43 enrolled patients. The therapeutic efficacy, complications, and follow-up results were assessed. RESULTS: The rate of en bloc resection was 100% (58/58), and en bloc resection with tumor-free lateral/basal margins (R0 resection) was 78% (45/58). There was no evidence of significant bleeding. Perforation occurred in 4 (6.9%) patients during the ESD, who were managed by conservative medical treatments after endoscopic closure of the perforation. Removal of 9 (16%) lesions resulted in esophageal stricture requiring balloon dilation after ESD. Of 40 lesions occurring in 31 patients fulfilling the criteria of node-negative tumors (mean follow-up, 17 months), 1 lesion resected by en bloc resection with nonevaluable tumor-free lateral margins (Rx [lateral] resection) recurred locally 6 months after ESD, which was treated successfully by a second ESD procedure. CONCLUSIONS: The ESD is applicable to the esophagus with promising results, but notification of risk is essential. PMID- 16713747 TI - A single molecule detection method for understanding mechanisms of electric field mediated interstitial transport of genes. AB - The interstitial space is a rate limiting physiological barrier to non-viral gene delivery. External pulsed electric fields have been proposed to increase DNA transport in the interstitium, thereby improving non-viral gene delivery. In order to characterize and improve the interstitial transport, we developed a reproducible single molecule detection method to observe the electromobility of DNA in a range of pulsed, high field strength electric fields typically used during electric field-mediated gene delivery. Using agarose gel as an interstitium phantom, we investigated the dependence of DNA electromobility on field magnitude, pulse duration, pulse interval, and pore size in the interstitial space. We observed that the characteristic electromobility behavior, exhibited under most pulsing conditions, consisted of three distinct phases: stretching, reptation, and relaxation. Electromobility depended strongly on the field magnitude, pulse duration, and pulse interval of the applied pulse sequences, as well as the pore size of the fibrous matrix through which the DNA migrated. Our data also suggest the existence of a minimum pulse amplitude required to initiate electrophoretic transport. These results are useful for understanding the mechanisms of DNA electromobility and improving interstitial transport of genes during electric field-mediated gene delivery. PMID- 16713749 TI - Mechanism of actin polymerization by myosin subfragment-1 probed by dynamic light scattering. AB - Monomeric actin (G-actin) polymerizes spontaneously into helical filaments in the presence of inorganic salts. The slowest, rate-limiting step of the polymerization process is formation of actin trimers, the smallest oligomers that serve as nuclei for fast filament growth (filament elongation) by monomer addition at the filament ends. In low ionic-strength solutions, actin can be polymerized by myosin subfragment-1 (S1). In early works it has been suggested that G-actin-S1 1:1 complexes (GS) assemble into filaments according to the nucleation-filament elongation scheme. Subsequent studies indicated that one S1 molecule can bind two actin monomers, and that oligomerization of the initial complexes is a fast reaction. This has led to suggest an alternative mechanism, with a ternary G(2)S complex and its oligomers being predominant intermediates of S1-induced assembly of G-actin into filaments. We used dynamic light scattering to analyze the initial steps of S1-induced polymerization of actin. Our results suggest formation of GS complexes and their oligomers in the presence of S1 equimolar to or in excess over actin. We confirm formation of G(2)S complexes as intermediates of S1-induced polymerization in the presence of actin in excess over S1. PMID- 16713748 TI - Determination of the lipid bilayer breakdown voltage by means of linear rising signal. AB - Electroporation is characterized by formation of structural changes within the cell membrane, which are caused by the presence of electrical field. It is believed that "pores" are mostly formed in lipid bilayer structure; if so, planar lipid bilayer represents a suitable model for experimental and theoretical studies of cell membrane electroporation. The breakdown voltage of the lipid bilayer is usually determined by repeatedly applying a rectangular voltage pulse. The amplitude of the voltage pulse is incremented in small steps until the breakdown of the bilayer is obtained. Using such a protocol each bilayer is exposed to a voltage pulse many times and the number of applied voltage pulses is not known in advance. Such a pre-treatment of the lipid bilayer affects its stability and consequently the breakdown voltage of the lipid bilayer. The aim of this study is to examine an alternative approach for determination of the lipid bilayer breakdown voltage by linear rising voltage signal. Different slopes of linear rising signal have been used in our experiments (POPC lipids; folding method for forming in the salt solution of 100 mM KCl). The breakdown voltage depends on the slope of the linear rising signal. Results show that gently sloping voltage signal electroporates the lipid bilayer at a lower voltage then steep voltage signal. Linear rising signal with gentle slope can be considered as having longer pre-treatment of the lipid bilayer; thus, the corresponding breakdown voltage is lower. With decreasing the slope of linear rising signal, minimal breakdown voltage for specific lipid bilayer can be determined. Based on our results, we suggest determination of lipid bilayer breakdown voltage by linear rising signal. Better reproducibility and lower scattering are obtained due to the fact that each bilayer is exposed to electroporation treatment only once. Moreover, minimal breakdown voltage for specific lipid bilayer can be determined. PMID- 16713750 TI - Optimization of the biological component of a bioelectrochemical cell. AB - The efficiency of electron transduction by Shewanella oneidensis MR-1 was investigated both in batch culture and in a dual-chambered electrochemical cell. Aerobically grown bacteria were inoculated into an insoluble FeOOH suspension in an anaerobic environment. As the bacteria reduced Fe(III) to Fe(II) there was a visible color change from red to bluish black; this simple color change assay proved to be a robust method for determining the electrochemical activity of S. oneidensis MR-1. In an effort to improve electricity production by S. oneidensis MR-1, the performance of the electrochemical cell with lactate as a fuel was first optimized with respect to both poised potential and fuel concentration. Ultimately, electricity production proved to be proportional to both the poised potential and to fuel concentration. In particular, higher poised potentials increased charge production. Finally, we attempted to optimize the bacteria themselves for the efficient transduction of reduced carbon sources into electricity. The batch culture underwent a series of serial dilutions; after the 4th dilution the microbe population exhibited a 30% increase in charge production. We are further exploring whether this increase was due to metabolic adaptations or to genetic mutations, and examining additional ways to evolve electrogenic organisms. PMID- 16713751 TI - Electrochemical characterization of biosensor based on nitrite reductase and methyl viologen co-immobilized glassy carbon electrode. AB - Nitrite reductase (NiR, nitric-oxide: ferricytochrome c oxidoreductase, EC 1.7.2.1) and methyl viologen (MV) were co-immobilized on glassy carbon electrode (GCE, d=3 mm) by polymer entrapment, and the electrode was tested as an electrochemical biosensor for amperometric determination of nitrite. The immobilization was performed by sequential loading and drying of a homogeneous mixture of poly(vinyl alcohol) (PVA), NiR and MV, followed by poly(allylamine hydrochloride) (PAH) solution, and finally hydrophilic polyurethane (HPU) dissolved in chloroform. The positively charged PAH layer could effectively keep immobilized cationic MV from diffusing through the membrane, holding mediator tightly near or on the electrode surface. The working principle of the biosensor was based on MV mediated electron transfer between electrode and immobilized NiR. The response time (t(90%)) of the biosensor was about 20 s and sensitivity was 11.8 nA/ microM (2.5 mU NiR) with linear response range of 1.5-260 microM (r(2)=0.996) and detection limit of 1.5 microM (S/N=3). Lineweaver-Burk plot showed that Michaelis-Menten constant (K(m,app)) was about 770 microM. The biosensor showed durable storage stability for 24 days (stored in ambient air at room temperature) retaining 80% of its initial activity, and showed satisfactory reproducibility (relative standard deviation (R.S.D.)=3.8%, n=9). Interference study showed that chlorate, chloride, sulfite, sulfate did not interfere with the nitrite determination, however, nitrate interfered with the determination with relative sensitivity of 38% (ratio of sensitivity for nitrate to that for nitrite). In addition to the full characterization of the biosensor, kinetic study was also conducted in solution and the homogeneous rate constant (k(2)) between NiR and MV were determined by chronoamperometry to be 5.8 x 10(5) M(-1) s(-1). PMID- 16713752 TI - Redox sensing by Escherichia coli: effects of copper ions as oxidizers on proton coupled membrane transport. AB - Escherichia coli is able to grow under anaerobic fermentation conditions upon a decrease in redox potential (E(h)). Indeed, upon a transition of E. coli MC4100 wild-type culture to stationary growth phase a decrease in E(h) from the positive values ( approximately +100 mV) to the negative ones ( approximately -520 mV) was observed, the acidification of the medium and the H(2) production were obtained. An oxidizer, copper ions (Cu(2+)) affected a bacterial growth in a concentration dependent manner (of 0.1 mM to 10 mM) increasing latent (lag) growth phase duration, delaying logarithmic (log) growth phase and decreasing specific growth rate. Acidification of the medium and the N,N'-dicyclohexylcarbodiimide (DCCD)- and azide-sensitive proton-potassium exchange by bacteria were inhibited, H(2) production upon growth and under assays disappeared with Cu(2+) (0.1 mM). These effects were observed with hycE but not hyfR and hyc(A-H) mutants and under aerobic conditions. Cu(2+) also increased membrane proton conductance. Copper ions are suggested to affect directly the F(0)F(1)-ATPase associated with potassium uptake transport system and/or formate hydrogenlyase composed with hydrogenase 4. A role of the F(0)F(1)-ATPase in redox sensing under fermentation is proposed. PMID- 16713753 TI - Trans fatty acids and cardiovascular disease-epidemiological data. AB - Prospective epidemiologic studies and case-control studies using adipose tissue analyses support a major role of trans fatty acids (TFA) in risk of coronary heart disease (CHD). The magnitude of the association with CHD is considerably stronger than for saturated fat, and is stronger than predicted by the effects of TFA on LDL and HDL cholesterol. The apparent gap between the epidemiologic findings and effects of TFA on LDL:HDL has been bridged, at least in part, by recent metabolic studies showing effects of TFA on inflammatory factors and other indicators of insulin resistance. PMID- 16713755 TI - Females with PDHA1 gene mutations: a diagnostic challenge. AB - Biochemical analysis was performed in muscle tissue and in fibroblasts of four unrelated females consecutively diagnosed with a 'de novo' point mutation in the PDHA1 gene. Pyruvate dehydrogenase E1 subunit deficiency was confirmed in the muscle sample of all patients, however, in three out of four cases the activity of the pyruvate dehydrogenase complex in fibroblasts showed a normal activity. A skewed inactivation was confirmed of the maternal X chromosome in fibroblasts in all children. Due to the possibility of a skewed X inactivation pattern enzyme measurements in fibroblasts are not always reliable for the diagnosis of a PDHc defect in females. Based on the overlapping features of PDHc deficiency with those of the disorders of the oxidative phosphorylation we suggest performing a fresh muscle biopsy for detailed biochemical analysis in females with a suspected pyruvate dehydrogenase deficiency, followed by molecular genetic analysis of the PDHA1 gene. PMID- 16713756 TI - Trapping of DNA topoisomerase I on nick-containing DNA in cell free extracts of Saccharomyces cerevisiae. AB - The aim of the present study was to identify proteins that bind nicked DNA intermediates formed in the course of base excision repair (BER) in cell free extracts of Saccharomyces cerevisiae. In mammalian cells, nicks in DNA are targets of proteins such as PARP-1 or XRCC1 that have no homologues in yeast. One of the most promising methodologies to trap proteins that interact with damaged DNA lies in using a photocrosslinking technique with photoactivable dNTP analogues such as exo-N-{2-[N-(4-azido-2,5-difluoro-3-chloropyridine-6-yl)-3 aminopropionyl]-aminoethyl}-2'-deoxycytidine-5'-triphosphate (FAP-dCTP) for enzymatic synthesis of DNA probes with a photoreactive dNMP residue at the 3' margin of a nick. Using this approach, we identified a major covalent DNA-protein adduct between a nick-containing 34-mer DNA duplex and a protein of a molecular mass of around 100-kDa. Unexpectedly, the formation of the 100-kDa adduct did not require the incorporation of the photoreactive dNMP residue at the 3'-margin of the nick nor exposure to near UV-light. However, the formation of the 100-kDa adduct strictly required a nick or a short gap in the DNA probe. Furthermore, the 100-kDa adduct was not detected in yeast extracts lacking DNA topoisomerase I (Top1). To further establish the nature of crosslinked protein, yeast Top1 was tagged with a Myc-epitope. In this case, the mobility of the Top1-DNA adduct increased by 7- kDa. Therefore, our data speak in favor of Top1 trapping by nicked DNA. In support of this hypothesis, purified yeast Top1 was also crosslinked to nicked DNA structures. Undamaged, uracil- and abasic (AP) site containing DNAs were unable to trap Top1 under the same assay conditions. Since nicked DNA structures are frequently formed in the course of BER, their covalent linkage to Top1 has the potential to interfere with BER in vivo. PMID- 16713754 TI - Ceramide forms channels in mitochondrial outer membranes at physiologically relevant concentrations. AB - Recent evidence suggests that the ability of ceramides to induce apoptosis is due to a direct action on mitochondria. Mitochondria are known to contain enzymes responsible for ceramide synthesis and hydrolysis and mitochondrial ceramide levels have been shown to be elevated prior to the mitochondrial phase of apoptosis. Ceramides have been reported to induce the release of intermembrane space proteins from mitochondria, which has been linked to their ability to form large channels in membranes. The aim of this study was to determine if the membrane concentration of ceramide required for the formation of protein permeable channels is within the range that is present in mitochondria during the induction phase of apoptosis. Only a very small percentage of the ceramide actually inserts into the mitochondrial membranes. The permeability of the mitochondrial outer membrane correlates directly with the level of ceramide in the membrane. Importantly, the concentration of ceramide at which significant channel formation occurs is consistent with the level of mitochondrial ceramide that occurs during the induction phase of apoptosis (4 pmol ceramide/nanomole phospholipid). Similar results were obtained with short- and long-chain ceramide. Ceramide channel formation is specific to mitochondrial membranes in that no channel formation occurs in the plasma membranes of erythrocytes even at concentrations 20 times higher than those required for channel formation in mitochondrial outer membranes. Thus, ceramide channels are good candidates for the pathway by which proapoptotic proteins are released from mitochondria during the induction phase of apoptosis. PMID- 16713757 TI - Cricothyroid muscle electrical activity during respiration and apneas in lambs. AB - Respiratory function of the cricothyroid muscle (CT) is virtually unknown in the neonatal period. This study was aimed at assessing CT electrical activity (EMG) during respiration and central apneas in non-sedated lambs. Seven full-term and four preterm lambs were instrumented for polysomnographic recording, including EMG of the diaphragm, thyroarytenoid (TA, a glottal constrictor), posterior cricoarytenoid (PCA, the primary glottal dilator) and CT. Phasic CT EMG was usually observed during inspiration and late expiration, whereas phasic TA EMG was observed during early expiration. While TA EMG virtually disappeared in REM sleep, both inspiratory and expiratory CT EMG increased. Overall, while CT EMG was not frequently observed during central apneas in either full-term (10% of apneas, but never simultaneously with TA EMG) or preterm lambs (30% of apneas), it was associated with decreased lung volume and subglottal pressure when present alone or with PCA EMG. Our results concur with the assumption that CT behaves as a laryngeal dilator in the neonatal period. PMID- 16713758 TI - Kinetic stability plays a dominant role in the denaturant-induced unfolding of Erythrina indica lectin. AB - The urea-induced denaturation of dimeric Erythrina indica lectin (EIL) has been studied at pH 7.2 under equilibrium and kinetic conditions in the temperature range of 40-55 degrees C. The structure of EIL is largely unaffected in this temperature range in absence of denaturant, and also in 8 M urea after incubation for 24 h at ambient temperature. The equilibrium denaturation of EIL exhibits a monophasic unfolding transition from the native dimer to the unfolded monomer as monitored by fluorescence, far-UV CD, and size-exclusion FPLC. The thermodynamic parameters determined for the two-state unfolding equilibrium show that the free energy of unfolding (DeltaGu, aq) remains practically same between 40 and 55 degrees C, with a value of 11.8 +/- 0.6 kcal mol(-1) (monomer units). The unfolding kinetics of EIL describes a single exponential decay pattern, and the apparent rate constants determined at different temperatures indicate that the rate of the unfolding reaction increases several fold with increase in temperature. The presence of probe like external metal ions (Mn2+, Ca2+) does not influence the unfolding reaction thermodynamically or kinetically; however, the presence of EDTA affects only kinetics. The present results suggest that the ability of EIL to preserve the structural integrity against the highly denaturing conditions is linked primarily to its kinetic stability, and the synergic action of heat and denaturant is involved in the unfolding of the protein. PMID- 16713759 TI - Pain control and sedation. PMID- 16713760 TI - Mechanisms of carrier-mediated intracellular delivery of therapeutics. Proceedings from a satellite meeting of the joint 15th IUPAB and EBSA International Biophysics Congress. August 25-26, 2005. PMID- 16713761 TI - Adolescent medicine. PMID- 16713762 TI - Office practice of adolescent medicine. AB - Physicians have an important role to play in the health of teenagers who come to their offices. The physician can help teenagers to understand their physical and emotional changes, to talk about sensitive issues in the family,and to identify and reduce some of the risks of adolescent life. The best strategy is to screen each adolescent and offer brief interventions or advice. A referral or a follow up visit may be necessary for teenagers with more complicated problems. A number of tools can help a physician's office meet the needs of its teenagers: screening forms, parent materials, an explanation of confidentiality, a community resource list, and health education materials. Meeting the challenge of good adolescent care makes for bet-ter adolescent health and a more rewarding office practice. PMID- 16713763 TI - Outside the margins: youth who are different and their special health care needs. AB - Some adolescents have differences, not of their own choosing, that require different approaches to clinical assessment and care. These adolescents are often marginalized from their peer group, and of-ten isolated in general. Awareness of the differences in health risks that these youth face, and their challenges in the successful mastery of developmental tasks is important to the clinician who serves them. Health care professionals must avoid three things in their efforts to help these groups of young people: judging, preaching, or becoming an agent of social control, even though such efforts maybe well meaning. This article addresses some of the unique needs of gay, lesbian, and bisexual youth; homeless adolescents; foster care youth; and youth who have disabilities. PMID- 16713764 TI - School-based health centers and the primary care physician: an opportunity for collaborative care. AB - Located throughout the country, SBHCs are an important resource for primary care physicians caring for children and adolescents. It is beneficial for primary care physicians to become familiar with school-based resources in their community. Information about SBHCs nationwide is available from the NASBHC [4,16] and The Center for Health and Health Care in Schools [3,7,12,15]. Partnering with SBHCs can have important benefits for your practice, including improved continuity of care, management of chronic dis-eases like asthma, opportunities for group and multidisciplinary interventions for overweight children, and access to SBHC mental health services. Specializing in adolescent issues, with the advantage of proximity, SBHC providers can be an invaluable asset in managing and helping to monitor your patients, especially when other community resources are limited or services are difficult to access. In our own centers, we are currently exploring ways to simplify ongoing routine communication with primary care providers. Consider discussing this opportunity for collaborative services with your patients and contacting your local SBHC to discuss how you would like to communicate and work together. PMID- 16713765 TI - Risky business: what primary care practitioners need to know about the influence of the media on adolescents. AB - Media can be powerfully prosocial, educational, and entertaining. Negative effects do exist, however, and they have been well documented. Despite the power of the new multinational media conglomerates and the intransigence of the entertainment industry,practitioners can make a difference in the future. PMID- 16713766 TI - Office counseling for the adolescent. AB - Physicians can and do influence adolescents' health-related behaviors. A recent study of low-income adolescents found that most adolescents report they are not receiving sufficient counseling about risk and risky behaviors from their physicians [8]. The results of this study and the author's personal experience (teaching physicians how to counsel and interview adolescents) support the need for physicians to learn more about office-based counseling with adolescents. Physicians who care for teens should learn how to meet the unique needs of individuals at varying stages of development and be prepared to offer counseling services tailored to the special needs of this population. Awareness of counseling techniques and knowledge of how to use those techniques effectively should help family medicine physicians to provide effective comprehensive health care to adolescent patients. PMID- 16713768 TI - Epidemiology and prevention of unintended pregnancy in adolescents. AB - Recent declines in adolescent pregnancy are encouraging, but current rates are still too high. Pregnancy prevention is a complex issue in the adolescent population, but much has been learned about effective interventions. Clinicians can and must play a key role in adolescent pregnancy prevention. PMID- 16713767 TI - Adolescent sexuality. AB - Adolescence is the period of life in which issues of sexuality come to the fore. This article describes the concept of sexuality, teens' expression of sexuality, and various consequences of sexual behaviors. Issues of sexual preference and identity are described,and a brief discussion of sex education is presented. Primary care clinicians should use this article as a basis for further exploration of these important issues for their adolescent patients. PMID- 16713769 TI - Contraceptive care for the adolescent. AB - Despite recent improvements in rates of teen pregnancy, abortion, birth,and contraceptive use, effective contraceptive counseling for adolescents should be a high priority for the primary care provider in the office setting. Adolescent psychosocial risk screening and appropriate counseling about sexual decision making is necessary. Contraceptive services visits include, relevant history, limited physical examination, provision of information, anticipatory guidance about sexual behaviors, and the provision of contraceptive methods. Teens should always be encouraged to use a male condom during sex to reduce STI risk. A variety of barrier and hormonal contraceptive methods are available for the adolescent population. Education about and provision of EC is effective in reducing the rate of unintended pregnancy and abortion in the United States. Contraceptive care for adolescents is a rewarding experience for primary care providers. PMID- 16713770 TI - Sexually transmitted infections: screening, syndromes, and symptoms. AB - STI are among the most common pathological conditions affecting sexually active adolescents. Both screening of asymptomatic adolescents and a thoughtful, thorough approach to the evaluation of symptoms are critical measures in STI identification and control. PMID- 16713771 TI - Breast disorders in children and adolescents. AB - Concerns about problems of the breast are often noted in adolescents and their parents. This review discusses issues and disorders of the breast in children and adolescents, starting with basic principles of embryology and adolescent breast development. Concepts that are covered include congenital breast disorders, abnormal timing of breast development, breast asymmetry, underdeveloped breasts, breast atrophy, tuberous breasts, mammary hyperplasia, fibroadenoma, giant fibroadenoma, cystosarcoma phyllodes, intra-ductal breast papilloma, adenocarcinoma, mastitis, traumatic breast disorders, benign breast disease, fibrocystic change, mastalgia, gynecomastia, and galactorrhea. A number of figures are provided illustrating breast pathologic findings. Clinicians caring for children and adolescents are encouraged to provide careful assessments of this important organ. PMID- 16713772 TI - Dysfunctional uterine bleeding in adolescents: concepts of pathophysiology and management. AB - DUB is extremely common in young adolescents, and can be regarded asa part of normal physiologic development. It is essential to have a firm grasp on normal physiologic development of the menstrual cycle to recognize the normal menstrual patterns of adolescents and to manage abnormal patterns that may develop. Specific management depends on the underlying cause, presence and extent of any existing anemia, and duration of the irregular menstrual pattern. PMID- 16713773 TI - The adolescent and substance use, an approach to office management. AB - Adolescence is a period of transition. During this time young people explore many options in an attempt to define who they are. Substance use or abuse is an area of experimentation that creates anxiety for everyone involved and in which, as a health care provider, a physician may be asked to intervene. This article provides general principles regarding substance use, information about commonly used drugs, and recommendations on the diagnosis and management of this problem in an office setting. PMID- 16713774 TI - Psychopharmacology for the violent adolescent. AB - Psychotropic medications for the violent adolescent can be used safely and effectively as long as a thorough evaluation is conducted beforehand,and appropriate monitoring is attended to for the duration of treatment. Table 1 summarizes several clinical conditions associated with aggressive and violent behaviors, and the recommendations for first-, second-, and third-line pharmacotherapeutic agents to treat them. PMID- 16713775 TI - Musculoskeletal injuries in sports. AB - A large number of adolescents participate in various sports. Not withstanding the methodologic problems with epidemiologic data, a large percentage of athletes sustain musculoskeletal injuries. In most instances, the athlete first presents to his or her primary care physician, who must perform the initial assessment and decide on further management. Many injuries can be managed by the primary care physician. It is important to recognize the unique characteristics of adolescent growth and development that have implications for the diagnosis and management of musculoskeletal injuries. PMID- 16713776 TI - Office management of the adolescent with diabetes mellitus. AB - Diabetes mellitus (DM) is a chronic metabolic disorder caused by absolute or relative insulin deficiency. It causes disturbances in carbohydrate, protein, and fat metabolism. Environmental triggers superimposed on a genetic predisposition are responsible for the clinical expression of the syndrome. This article outlines current ideas in diagnosis and treatment of diabetes in adolescents. The"team approach" to outpatient diabetes management is described,with the hope that it will be of benefit to primary care physicians and their office staff. Advances in diabetes care regimens, and tools used in diabetes care are briefly summarized. Summary tables are provided to assist in routine care, troubleshooting, and advanced management. PMID- 16713777 TI - Economic value of out-of-hospital emergency care: a structured literature review. AB - STUDY OBJECTIVE: The evaluation of the impact of out-of-hospital emergency care is a relatively new research focus. As such, there is a compelling need to determine how finite health care resources should be used in this setting. The objective of this study is to conduct a structured review of published economic evaluations of out-of-hospital emergency care to assess its economic value. METHODS: A structured literature search and structured review of articles pertaining to the economic value of out-of-hospital care was performed. The bibliographic database MEDLINE was searched for pertinent English-language articles published between 1966 and 2003. The search used the medical subject headings "emergency medical services" and "emergency medical technician" and was limited to the subheading "economics" and crossed with the medical subject heading "economics." The titles generated by this search were systematically reviewed and limited by topic. Abstracts from the identified titles were reviewed to select a final set of pertinent articles. These articles were further limited based on explicit inclusion and exclusion criteria. Authors used a previously published structured evaluation tool to review the final set of identified articles for quality and content. RESULTS: The initial MEDLINE search identified 3,533 citations. From this set, 535 potentially relevant abstracts were reviewed. From the abstract review, 46 articles were identified, along with an additional 14 from searching the secondary references. Of these 60 articles, 32 met the review inclusion criteria and were subjected to a full structured review. These studies predominantly addressed the cost of cardiac arrest (n=13, 41%), major trauma (n=8, 25%), and emergency medical services treatment in general (n=8, 25%). Only 14 studies considered the costs and consequences of competing alternatives. Of these, 2 were cost-benefit and 12 were cost-effectiveness evaluations. Two of the 14 studies met all 10 criteria for high-quality economic evaluation, whereas 2 others met none. CONCLUSION: There is a paucity of out-of hospital care literature that addresses cost and economic value. The extant literature is limited in scope, poor in quality, and evaluates small subsets of out-of-hospital emergency care costs. Favorable cost-effectiveness has not been firmly established for most aspects of out-of-hospital emergency care. PMID- 16713778 TI - Etomidate versus midazolam for out-of-hospital intubation: a prospective, randomized trial. AB - STUDY OBJECTIVE: The primary objective of this study is to compare the intubation success rates of etomidate and midazolam when used for sedative-facilitated intubation, without paralytics, in out-of-hospital adult patients. METHODS: This prospective, double-blind, randomized trial was conducted with 2 ground out-of hospital advanced life support systems (ALS); all patients aged 18 or older who required out-of-hospital sedative-facilitated intubation were eligible for participation. The ambulances were stocked with blinded numbered syringes containing either 7 mg of midazolam or 20 mg of etomidate. No paralytics were used. If sedation was not achieved with the study drug, medics could request additional sedation from a medical command physician; only midazolam or diazepam were available outside of the study. RESULTS: One hundred ten patients were enrolled in the study; 55 patients received midazolam and 55 patients received etomidate. The 2 groups were similar with regard to age, sex, initial vital signs, and reasons for intubation or sedation. The overall intubation success rate was 76% (95% confidence interval [CI] 68% to 84%); 75% (41/55) for midazolam (95% CI 64% to 86%) and 76% (42/55) for etomidate (95% CI 65% to 87%). There was also no difference in incidence of hypotension, number of intubation attempts, or perceived difficulty of intubation. Additional sedation was requested almost equally for the 2 groups: 14 patients in the midazolam group and 12 patients in the etomidate group. A benzodiazepine was successful for rescue of a failed etomidate intubation 10 of 12 times (83%; 95% CI 62% to 100%). When used for rescue of failed midazolam intubations, benzodiazepines were effective in only 5 of 14 (36%, 95% CI 11% to 61%) attempts. CONCLUSION: There were no observed differences between midazolam and etomidate in sedation-facilitated intubation success rates; we could not fully evaluate global outcomes of these agents or the sedative-facilitated intubation strategy itself. PMID- 16713779 TI - Images in emergency medicine. Bullous disease of dialysis. PMID- 16713780 TI - Out-of-hospital endotracheal intubation: where are we? AB - While remaining prominent in paramedic care and beneficial to some patients, out of-hospital endotracheal intubation has not clearly improved survival or reduced morbidity from critical illness or injury when studied more broadly. Recent studies identify equivocal or unfavorable clinical effects, adverse events and errors, interaction with other important resuscitation interventions, and challenges in providing and maintaining procedural skill. We provide an overview of current data evaluating the overall effectiveness, safety, and feasibility of paramedic out-of-hospital endotracheal intubation. These studies highlight our limited understanding of out-of-hospital endotracheal intubation and the need for new strategies to improve airway support in the out-of-hospital setting. PMID- 16713781 TI - Out-of-hospital endotracheal intubation: half empty or half full? PMID- 16713783 TI - Intracuff pressures of endotracheal tubes in the management of airway emergencies: the need for pressure monitoring. AB - STUDY OBJECTIVE: Excessive pressure exerted on the tracheal mucosa is an avoidable factor implicated as a cause of damage after intubation of the trachea with cuffed tubes. Many patients are intubated in the out-of-hospital setting by emergency medical teams. The time spent in the out-of-hospital setting could very well be long enough for tracheal mucosal damage to occur if cuff pressure is not controlled. The objective of this study is to assess the incidence of intracuff excessive pressure in the out-of-hospital setting. METHODS: We performed an observational prospective study. Every patient who required tracheal intubation was included in the study, regardless of indication. When the patient was stabilized, the cuff was connected to a manometer, and pressure was systematically recorded. Corrections to inflation were performed if necessary to achieve a cuff pressure of 14 to 27 cm H2O. RESULTS: One hundred seven patients were included. Eighty-five were out-of-hospital patients and 22 were transfers between 2 hospitals who had been previously intubated when the mobile intensive care unit team arrived. The first recorded cuff pressures were greater than 27 cm H2O among 79% of patients (85/107), with a mean pressure of 56 cm H2O (SD+/-34 cm H2O) in out-of-hospital patients and 69 cm H2O (SD+/-37 cm H2O) for transferred patients. Pressure correction was made in 72% of patients (77/107). There were corrections in 69% (59/85) of out-of-hospital patients and 82% (18/22) of transferred patients. CONCLUSION: This study revealed that the majority of cuff pressures exceeded safe pressure and required correction. Frequent measurement and adjustment of cuff pressure has been recommended, but this method requires a specific manometer. PMID- 16713784 TI - Laryngeal view during laryngoscopy: a randomized trial comparing cricoid pressure, backward-upward-rightward pressure, and bimanual laryngoscopy. AB - STUDY OBJECTIVE: External cricoid and thyroid cartilage manipulations are commonly taught to facilitate laryngeal view during intubation. We compare the laryngeal views during laryngoscopy with 4 manipulations (no manipulation, cricoid pressure, backward-upward-rightward pressure [BURP], and bimanual laryngoscopy) to determine the method that optimizes laryngeal view. METHODS: This was a randomized intervention study involving emergency physicians participating in airway training courses from December 2003 to November 2004. Direct laryngoscopies were performed with curved blades on fresh, non-fixed cadavers by using each of the 4 methods. The percentage of glottic opening (POGO), a validated scoring scale, was recorded for each laryngoscopy. Scores for bimanual laryngoscopy were recorded before the assistant applied external pressure. RESULTS: A total of 1,530 sets of comparative laryngoscopies were performed by 104 participants. One thousand one hundred eighteen of 1,530 sets (73%) had POGO scores less than 100 with no manipulation. Compared to no manipulation, mean POGO scores with bimanual laryngoscopy improved by 25 (95% confidence interval [CI] 23 to 27); mean POGO score improvement with cricoid pressure and BURP were 5 (95% CI 3 to 8) and 4 (95% CI 1 to 7), respectively. POGO scores with bimanual laryngoscopy were higher compared to cricoid pressure (mean difference 20, 95% CI 17 to 22) and BURP (mean difference 21, 95% CI 19 to 24). Among laryngoscopies with no manipulation in which the POGO score greater than 0 (n=1,434), laryngeal view worsened in 60 cases (4%, 95% CI 3% to 5%) with bimanual laryngoscopy, in 409 cases (29%, 95% CI 26% to 31%) with cricoid pressure, and in 504 cases (35%, 95% CI 33% to 38%) with BURP. CONCLUSION: Using a cadaver model, we found pressing on the neck during curved blade laryngoscopy greatly affects laryngeal view. Overall, bimanual laryngoscopy improved the view compared to cricoid pressure, BURP, and no manipulation. Cricoid pressure and BURP frequently worsen laryngoscopy. These data suggest bimanual laryngoscopy should be considered when teaching emergency airway management. PMID- 16713785 TI - Adverse events associated with the use of intravenous epinephrine in emergency department patients presenting with severe asthma. AB - STUDY OBJECTIVE: We determine the rate of adverse effects associated with the use of intravenous (IV) epinephrine by infusion for the treatment of severe asthma in the emergency department (ED). METHODS: This retrospective, structured, medical record review included adult patients who presented to the ED of Western Hospital between 1998 and 2003 and who were triaged as category 1, 2, or 3, had a discharge diagnosis of asthma, and were administered IV epinephrine in the ED. Patients were excluded if they were older than 55 years or if a diagnosis of asthma was not confirmed. The primary outcome measures were occurrence of cardiac arrhythmia or ischemia, local tissue ischemia, hypotension or hypertension, neurologic injury, or death related to epinephrine infusion. RESULTS: Two hundred twenty episodes of care met the inclusion criteria. Adverse events occurred in 67 episodes (30.5%; 95% confidence interval [CI] 24.5% to 37.1%); however, most were minor and self-limiting. There were no deaths. Major adverse events occurred in 3.6% of cases (8/220; 95% CI 1.7% to 7.3%), including 2 cases of supraventricular tachycardia, 1 case of chest pain with ECG changes, 1 case of incidental elevated troponin, and 4 cases of hypotension requiring intervention. CONCLUSION: IV epinephrine is associated with a low rate of major and a moderate rate of minor adverse events in patients with severe asthma; however, a causal relationship has not been established. Further research investigating effectiveness, as well as safety, is warranted. PMID- 16713786 TI - Emergency department treatment of severe acute asthma. PMID- 16713787 TI - Lawnmower injuries in the United States: 1996 to 2004. AB - STUDY OBJECTIVE: We update the epidemiology of lawnmower injuries, together with leading mechanisms of lawnmower injury in the United States, for the entire age range by using nationally representative data. METHODS: Data were obtained from the National Hospital Discharge Survey 1996-2003 and the National Electronic Injury Surveillance System 1996-2004. RESULTS: Individuals in the 60- to 69-year age group had the highest push mower injury incidence in 2004, whereas those in the 70 years and older age group had the highest riding mower injury incidence. Children younger than 15 years also had a substantial injury incidence. Individuals in the 15- to 19-year age group had the highest rate of hospitalizations caused by lawnmower injuries from 1996 through 2003, with 0.72 per 100,000 person-years (95% confidence interval 0.07 to 1.36). Debris from under the mower hitting a body part or entering the eye was the most common mechanism for lawnmower injury. The second most common mechanism of injury was nonspecific pain onset after the ordinary operation of the mower. Fracture of 1 or more phalanges of the foot was the most common diagnosis among lawnmower injury hospitalizations, with 34.4%, followed by traumatic amputation of the toe, with 32.4%. There is an increasing trend of lawnmower injuries in the United States during the last 9 years. CONCLUSION: Lawnmower injuries increase with age, with peaks in persons older than 59 years. Given the high incidence of projectile related injuries, improved protective apparel and eyewear could lower the rate of injury for all age groups. The increasing trend of lawnmower injuries in the United States suggests that more must be done to prevent lawnmower injuries. PMID- 16713788 TI - National Highway Traffic Safety Administration (NHTSA) notes. Contrasting rural and urban fatal crashes 1994-2003. PMID- 16713790 TI - Images in emergency medicine. Ramsay Hunt syndrome: a rare entity. PMID- 16713791 TI - Blood cultures in pneumonia patients. PMID- 16713792 TI - Difficult airways are "LEMONS": updating the LEMON mneumonic to account for time and oxygen reserve. PMID- 16713795 TI - Emergency physician contractual relationships. PMID- 16713796 TI - Crowding. PMID- 16713797 TI - Public training in cardiopulmonary resuscitation and public access defibrillation. PMID- 16713798 TI - Patient medical condition identification systems. PMID- 16713799 TI - A time for perspective on bisphosphonates. PMID- 16713800 TI - Chronic oral inflammation and the progression of periodontal pathology in the third molar region. AB - PURPOSE: To assess the association between risk markers of chronic oral inflammation and changes over time in periodontal probing depth (PD) in the third molar region, the distal of a second molar, or around a third molar. SUBJECTS AND METHODS: The data from these analyses are part of a study of subjects enrolled with 4 asymptomatic third molars with adjacent second molars in an institutional review board-approved longitudinal trial. Full-mouth periodontal probing was conducted at enrollment and follow-up. Enrollment levels of periodontal pathogens and gingival crevicular fluid inflammatory mediators were assayed as indicators of the degree of oral inflammation. Subjects were categorized as those who had at least a 2 mm change in periodontal PD between baseline and follow-up in the third molar region and those who did not. The relationship between aggregated subject baseline PD, levels of periodontal pathogens, and gingival crevicular fluid IL-1 beta, and the proportion of subjects with changes in PD >or=2 mm versus those with PD <2 mm were compared with Cochran-Mantel-Haenzsel statistics. Level of significance was set at 0.05. Risk assessment models for a change in PD >or=2 mm were developed using logistic regression analysis. RESULTS: Twenty-four percent of 254 subjects exhibited a change in PD from baseline to follow-up of >or=2 mm in the third molar region. Of these, 95% had a baseline PD of >or=4 mm. Both high (>or=10(5)) "orange" and "red" complex bacteria and PD of >or=4 mm detected at enrollment were significantly associated with a change in PD >or=2 mm. Odds of a change in PD >or=2 mm were increased if baseline pathogen levels were >or=10(5) or a PD of >or=4 mm was detected at enrollment. CONCLUSION: Our findings are consistent with chronic oral inflammation leading to a progression of periodontal disease in the third molar region. PMID- 16713801 TI - Oral appliances and maxillomandibular advancement surgery: an alternative treatment protocol for the obstructive sleep apnea-hypopnea syndrome. AB - PURPOSE: The present study comprises a retrospective evaluation of the potential application of mandibular repositioning appliance (MRA) therapy preceding maxillomandibular advancement (MMA) surgery in the treatment of the Obstructive Sleep Apnea-Hypopnea Syndrome (OSAHS). Our initial experiences with a new surgical protocol, in which MRA therapy serves as a predictor for success of MMA surgery, are reported. PATIENTS AND METHODS: Forty-three consecutive patients with OSAHS treated with MRA therapy were considered for inclusion (mean+/-SD; Apnea-Hypopnea Index [AHI]=27+/-20; age=53+/-9 years). All patients displaying a substantial improvement in their AHI with MRA therapy (ie,>50% reduction) who preferred surgical rather than "prosthetic" advancement of the mandible were offered MMA surgery. Accordingly, 4 out of 43 patients were treated with MMA surgery. The AHI was used as the primary outcome measure, with MMA surgery being considered successful in case of a postoperative AHI <5. RESULTS: All 4 patients included displayed substantial improvement in their AHI following MRA therapy. Moreover, in 3 patients MRA therapy resulted in a post-treatment AHI 40 minutes). Using univariate and multivariate analysis, we compared self-reported physician visit time when antibiotics were and were not prescribed for viral RTIs. RESULTS: 2739 visits from the NAMCS database, representing 119,926 visits nationally, met study criteria. Antibiotics were prescribed at 46,949 (39%) visits-75% with a diagnosis of bronchitis, 54% with bronchiolitis, and 30% with cold or URI. After adjusting for factors related to physician visit time, there was no difference in visit duration when antibiotics were or were not prescribed (13.6 +/- 8.4 and 13.3 +/- 9.6 minutes, respectively, P = 0.24). CONCLUSION: While antibiotics prescribing for viral RTI in children occurred frequently, our findings do not support the contention that it takes longer 'not to prescribe' antibiotics for children with viral RTIs. PMID- 16713934 TI - Effectiveness of an educational intervention in oral health for pediatric residents. AB - OBJECTIVE: To evaluate the effectiveness of an oral health educational intervention designed to increase proficiency of pediatric residents in oral health. METHODS: Pediatric residents at the University of North Carolina at Chapel Hill (UNC) participated in oral health education that included didactic sessions, hands-on instruction by pediatric dentists and residents, preventive dentistry prompts, and change strategies to introduce oral health into practice. Pediatric residents at East Carolina University (ECU), who had a short practicum in oral health, and Wake Forest University (WFU), who had no specific oral health instruction, served as comparison groups. All residents completed questionnaires before and 12 months after instruction began at UNC. Effects were tested for each school separately by repeated-measure analysis of variance. RESULTS: The mean percentage of UNC residents who answered 18 knowledge questions correctly and reported frequently performing 10 preventive dental practices increased by 17.7% and 65.1%, respectively, from baseline levels. Residents' confidence in performing 10 counseling and oral health screening activities improved by 17.9%. Opinions about including oral health in their clinical care improved only slightly. Changes at ECU and WFU were small, but low response rates in those schools preclude substantive conclusions from between schools comparisons. CONCLUSIONS: Multifaceted instruction in oral health was effective in improving pediatric residents' knowledge about oral health, their confidence in providing oral health services, and the delivery of these services in their ambulatory care practices. Residents also adopted the use of fluoride varnish, an innovation in pediatrics. More studies are needed to define the most efficient and effective residency-based instruction. PMID- 16713935 TI - Individually significant risk factors do not provide an accurate clinical prediction rule for infant underimmunization in one disadvantaged urban area. AB - OBJECTIVE: To define a clinical prediction rule for underimmunization in children of low socioeconomic status. METHODS: We assessed a cohort of 1160 infants born from July 1998 through June 1999 at an urban safety net hospital that received primary care at 4 community health centers. The main outcome measure was up-to date status with the 3:2:2:2 infant vaccine series at 12 months of age. RESULTS: Latino infants (n = 959, 83% of cohort) had immunization rates of 74%, at least 18% higher than any other racial/ethnic group. Multivariate logistic regression demonstrated the following independent associations (relative risk, 95% confidence interval) for inadequate immunization: non-Latino ethnicity (1.7, 1.4 2.0), maternal smoking (1.3, 1.1-1.7), no health insurance (1.9, 1.4-2.3), late prenatal care (1.9, 1.5-2.3), no pediatric chronic condition (2.1, 1.2-3.1), and no intent to breast-feed (1.3, 1.1-1.6). However, the index of concordance (c index) for this model was only 0.69. Neither excluding infants who left the health care system nor accounting for infants who were "late starters" for their first vaccines improved the predictive accuracy of the model. CONCLUSIONS: In this predominantly Latino population of low socioeconomic status, Latino infants have higher immunization rates than other infants. However, we were unable to develop a model to reliably predict which infants in this population were underimmunized. Models to predict underimmunization should be tested in other settings. In this population, interventions to improve immunization rates must be targeted at all children without respect to individual risk factors. PMID- 16713936 TI - The impact of inactive infants on clinic-based immunization rates. AB - OBJECTIVE: Clinic-based immunization rates are used to evaluate clinic performance and immunization interventions, but they typically exclude so-called inactive patients (ie, those who no longer receive care at the clinic). We assessed the effect of enhanced ascertainment of inactive patients on clinic based immunization rates and on the conclusions of a randomized controlled trial. METHODS: The study was a post hoc analysis of a cluster-randomized controlled trial. Infant randomization to the immunization intervention (4 clinics) or control group (4 clinics) was based on the site of their 2-week well-child care visit. The study was conducted at an integrated inner-city health care system serving a low-income population. A total of 2190 infants born between July 1, 1998, and June 30, 1999, who attended at least 1 postnatal visit, participated. In control sites, clinic staff documented inactive infants in the immunization registry and medical charts. The research staff undertook additional patient tracking efforts in the intervention clinics. RESULTS: Control clinics identified 155 (13.4%) of 1160 children as inactive within 1 year of birth, whereas 284 (27.6%) of 1030 intervention infants were documented as inactive (P < .001). In bivariate analyses from the randomized trial, immunization rates differed between intervention and control branches. In multivariate models, immunization rates were significantly higher in the intervention branch when inactive infants were removed (adjusted relative risk 1.58; 95% confidence interval, 1.28-1.89), but not when they were included (adjusted relative risk 1.09; 95% confidence interval, 0.97-1.21). CONCLUSIONS: Additional patient tracking efforts can dramatically influence inactive patient documentation and clinic-based immunization rates used for various purposes. PMID- 16713937 TI - Trends in diabetes mellitus among privately insured children, 1998-2002. AB - OBJECTIVE: To evaluate trends in health care administrative claims for childhood diabetes mellitus. METHODS: We conducted a serial cross-sectional study of a national sample of privately insured children < or = 18 years old. The number of subjects ranged from 306991 in 1998 to 974407 in 2002. We classified diabetes type by 2 schemes: one based on encounter claims only, and the other based on both encounter and pharmacy claims. The prevalence of diabetes was determined after adjusting for demographic changes in the study population, including age, proportion enrolled in managed care, and urban residence. RESULTS: The adjusted prevalence of diabetes overall increased from 183 (95% confidence interval [CI], 169-198) cases per 100,000 enrollees in 1998 to 218 (95% CI, 208-228) cases per 100,000 enrollees in 2002, primarily because of an increase in type 1 diabetes (based on both encounter and pharmacy claims, 135 [95% CI, 123-148] cases per 100000 enrollees in 1998 to 167 [95% CI, 158-176] cases per 100,000 enrollees in 2002). CONCLUSIONS: Our estimate of the overall prevalence of diabetes is consistent with national data. However, our finding that rising prevalence of type 1 diabetes appear to account for most of the increase in diabetes claims is surprising. Future research will be needed to validate these results. PMID- 16713938 TI - Mammoths. PMID- 16713939 TI - Bird evolution. PMID- 16713940 TI - Shared neural circuitry for female and male sexual behaviours in Drosophila. PMID- 16713941 TI - Gene expression becomes heterogeneous with age. PMID- 16713942 TI - Eye development: random precision in color vision. AB - In insect and vertebrate eyes, different types of color-detecting photoreceptors are randomly distributed throughout the retina. A recent study has provided important new insights into the developmental mechanisms that generate the retinal mosaic required for color vision. PMID- 16713943 TI - Bacterial cell biology: managing magnetosomes. AB - Sensing of magnetic fields by living organisms -- magnetosensing -- is best understood in magnetotactic bacteria. Recently work has provided new insight into the biogenesis of bacterial magnetosomes, and links these organelles to a newly recognized prokaryotic cytoskeletal filament which organizes magnetosomes into a sensory structure capable of aligning cells with the geomagnetic field. PMID- 16713944 TI - Social evolution: cooperation by conflict. AB - A recent study suggests that aggression between wasps depends upon the costs and benefits of fighting, as determined by the position of individuals in a dominance hierarchy. PMID- 16713945 TI - Vertebrate segmentation: snail counts the time until morphogenesis. AB - During segmentation of vertebrate embryos, unsegmented mesenchymal mesoderm is divided into epithelial segments called somites. This process is governed by oscillating gene expression of the somite clock. A recent paper identifies the transcription factor Snail as a link between the somite clock and the control of somite morphogenesis. PMID- 16713946 TI - Insect vision: remembering the shape of things. AB - How does the nervous system store a newly experienced visual pattern, and how is that pattern subsequently made available for recognition? Recent work in Drosophila suggests that specific pattern features are stored separately in the nervous system. PMID- 16713947 TI - Eukaryotic transcription: what does it mean for a gene to be 'on'? AB - Until recently, transcription could only be observed by measuring mRNA production of cell populations, thus obscuring the kinetics at the level of individual transcription events. A new study now shows that eukaryotic transcription, visualised in individual living cells, occurs in bursts -- much as it does in prokaryotes. PMID- 16713948 TI - Wiring specificity: axon-dendrite matching refines the olfactory map. AB - In Drosophila, about 50 classes of olfactory receptor neurons enter the brain where their axons form highly specific synapses with the dendrites of identified partner neurons. A recent study has shown that genetic manipulations that shift the position of one class of postsynaptic dendrites can cause an exact corresponding shift in the location of their partner axons. PMID- 16713949 TI - Biological motion: a perceptual life detector? AB - Detecting the direction of motion of other animals is critical for a variety of adaptive behaviours, including filial attachment and hunting prey. Two new studies support the view that some vertebrates, including humans, have primitive brain systems for the visual detection of other legged vertebrates. PMID- 16713950 TI - Transcription under the control of nuclear Arm/beta-catenin. AB - The Wingless/Wnt pathway controls cell fates during animal development and regulates tissue homeostasis as well as stem cell number and differentiation in epithelia. Deregulation of Wnt signaling has been associated with cancer in humans. In the nucleus, the Wingless/Wnt signal is transmitted via the key effector protein Armadillo/beta-catenin. The recent identification and functional analysis of novel Armadillo/beta-catenin interaction partners provide new and exciting insights into the highly complex mechanism of Wingless/Wnt target gene activation. PMID- 16713951 TI - oko meduzy and related crumbs genes are determinants of apical cell features in the vertebrate embryo. AB - BACKGROUND: Polarity is an essential attribute of most eukaryotic cells. One of the most prominent features of cell polarity in many tissues is the subdivision of cell membrane into apical and basolateral compartments by a belt of cell junctions. The proper formation of this subdivision is of key importance. In sensory cells, for example, the apical membrane compartment differentiates specialized structures responsible for the detection of visual, auditory, and olfactory stimuli. In other tissues, apical specializations are responsible for the propagation of fluid flow. Despite its importance, the role of genetic determinants of apico-basal polarity in vertebrate embryogenesis remains poorly investigated. RESULTS: We show that zebrafish oko meduzy (ome) locus encodes a crumbs gene homolog, essential for the proper apico-basal polarity of neural tube epithelia. Two ome paralogs, crb2b and crb3a, promote the formation of apical cell features: photoreceptor inner segments and cilia in renal and auditory systems. The motility of cilia is defective following the impairment of crb2b function. Apical surface defects in ome- and crb2b-deficient animals are associated with profound disorganization of neuronal architecture and with the formation of pronephric cysts, respectively. Unexpectedly, despite differences in their structure and expression patterns, crumbs genes are, at least partially, functionally interchangeable. CONCLUSIONS: ome and related crumbs genes are necessary for the formation of gross morphological features in several organs, including the CNS and the renal system. On the cellular level, crumbs genes regulate the formation of both ciliary and nonciliary apical membrane compartment. PMID- 16713952 TI - Functional genomics identifies TOR-regulated genes that control growth and division. AB - BACKGROUND: The TOR (target of rapamycin) ser/thr protein kinase is the central component of a eukaryotic signaling pathway that regulates growth and is the direct target of the clinically useful drug rapamycin. Recent efforts have identified at least two multiprotein complexes that contain TOR, but little is known in higher eukaryotes about the genes downstream of TOR that control growth. RESULTS: By combining the use of a small molecule inhibitor (rapamycin), transcriptional profiling, and RNA interference in Drosophila tissue culture cells, we identified genes whose expression responds to Drosophila TOR (dTOR) inhibition and that regulate cell size. Several of the dTOR-regulated genes that function in cell size control have additional roles in cell division. Most of these genes are conserved in mammals and several are linked to human disease. This set of genes is highly enriched for regulators of ribosome biogenesis, which emphasizes the importance of TOR-dependent transcription in building the protein synthesis machinery in higher eukaryotes. In addition, we identify two dTOR regulated genes, CG3071 and CG6677, whose human orthologs, SAW and ASH2L, are also under TOR-dependent transcriptional control and encode proteins with conserved functional roles in growth. CONCLUSIONS: We conclude that combining RNA interference with genomic analysis approaches, such as transcriptional profiling, is an effective way to identify genes functioning in a particular biological process. Moreover, this strategy, if applied in model systems with simpler genomes, can identify genes with conserved functions in mammals. PMID- 16713953 TI - The RNA methyltransferase Misu (NSun2) mediates Myc-induced proliferation and is upregulated in tumors. AB - BACKGROUND: Myc is a well-known proto-oncogene, but its functions in normal tissue remain enigmatic. In adult epidermis, Myc stimulates exit from the stem cell compartment, decreasing cell adhesion and, by an unknown mechanism, triggering proliferation of transit-amplifying cells. RESULTS: We describe a novel direct target gene of Myc, Misu, that is expressed at low levels in normal epidermis but is upregulated on Myc activation. Misu encodes a previously uncharacterized RNA methyltransferase with high sequence homology to NSun2 and defines a new family of mammalian SUN-domain-containing proteins. The nucleolar localization of Misu is dependent on RNA polymerase III transcripts, and knockdown of Misu decreases nucleolar size. In G2 phase of the cell cycle, Misu is found in cytoplasmic vesicles, and it decorates the spindle in mitosis. Misu expression is highest in S phase, and RNAi constructs block Myc-induced keratinocyte proliferation and cell-cycle progression. Misu is expressed at low levels in normal tissues, but is highly induced in a range of tumors. Growth of human squamous-cell-carcinoma xenografts is decreased by Misu RNAi. CONCLUSIONS: Misu is a novel downstream Myc target that methylates RNA polymerase III transcripts. Misu mediates Myc-induced cell proliferation and growth and is a potential target for cancer therapies. PMID- 16713954 TI - Normal mitochondrial dynamics requires rhomboid-7 and affects Drosophila lifespan and neuronal function. AB - In addition to being energy generators, mitochondria control many cellular processes including apoptosis. They are dynamic organelles, and the machinery of membrane fusion and fission is emerging as a key regulator of mitochondrial biology. We have recently identified a novel and conserved mitochondrial rhomboid intramembrane protease that controls membrane fusion in Saccharomyces cerevisiae by processing the dynamin-like GTPase, Mgm1, thereby releasing it from the membrane. The genetics of mitochondrial membrane dynamics has until now focused primarily on yeast. Here we show that in Drosophila, the mitochondrial rhomboid (Rhomboid-7) is required for mitochondrial fusion during fly spermatogenesis and muscle maturation, both tissues with unusual mitochondrial dynamics. We also find that mutations in Drosophila optic atrophy 1-like (Opa1-like), the ortholog of yeast mgm1, display similar phenotypes, suggesting a shared role for Rhomboid-7 and Opa1-like, as with their yeast orthologs. Loss of human OPA1 leads to dominant optic atrophy, a mitochondrial disease leading to childhood onset blindness. rhomboid-7 mutant flies have severe neurological defects, evidenced by compromised signaling across the first visual synapse, as well as light-induced neurodegeneration of photoreceptors that resembles the human disease. rhomboid-7 mutant flies also have a greatly reduced lifespan. PMID- 16713955 TI - Phenotypically linked dichotomy in sea turtle foraging requires multiple conservation approaches. AB - Marine turtles undergo dramatic ontogenic changes in body size and behavior, with the loggerhead sea turtle, Caretta caretta, typically switching from an initial oceanic juvenile stage to one in the neritic, where maturation is reached and breeding migrations are subsequently undertaken every 2-3 years. Using satellite tracking, we investigated the migratory movements of adult females from one of the world's largest nesting aggregations at Cape Verde, West Africa. In direct contrast with the accepted life-history model for this species, results reveal two distinct adult foraging strategies that appear to be linked to body size. The larger turtles (n = 3) foraged in coastal waters, whereas smaller individuals (n = 7) foraged oceanically. The conservation implications of these findings are profound, with the population compartmentalized into habitats that may be differentially impacted by fishery threats in what is a global fishing hotspot. Although the protection of discrete areas containing coastal individuals may be attainable, the more numerous pelagic individuals are widely dispersed with individuals roaming over more than half a million square kilometers. Therefore, mitigation of fisheries by-catch for sea turtles in the east Atlantic will likely require complex and regionally tailored actions to account for this dichotomous behavior. PMID- 16713956 TI - Rapid displacement of a monoecious plant lineage is due to pollen swamping by a dioecious relative. AB - Interspecific hybridization is recognized as a potentially destructive process that represents a major threat to biodiversity. The rate of population displacement by hybridization can be rapid, but underlying mechanisms are often obscure. One hypothesis is that a species may be driven to extinction by interspecific gene flow, or pollen swamping, when hybrids are inviable or sterile. Here, we document the rapid movement of two zones of contact between monoecious hexaploid and dioecious diploid populations of the wind-pollinated plant Mercurialis annua (Euphorbiaceae) in northeastern and northwestern Spain, where diploids have displaced hexaploids by about 80 and 200 km, respectively, over a period of four decades. By using experimental mating arrays, we show that hybridization is highly asymmetrical in favor of the diploids, mainly because they disperse substantially more pollen, as expected in a comparison between an obligate outcrosser and a facultative selfer. Self-fertilization, which is expected to reduce the proportion of sterile hybrids produced in mixed ploidy populations, allowed the hexaploids to avoid the effects of pollen swamping only slightly, and in a density-dependent manner. Our results thus provide a mechanistic explanation for the rapid movement of both contact zones of M. annua in Spain. PMID- 16713957 TI - Sleep-dependent consolidation of contextual learning. AB - Memory consolidation is facilitated by sleep. Specifying the functional domain of sleep-dependent consolidation (SDC) is important for identifying the neural mechanisms underlying this phenomenon. Previous work indicates that SDC may be limited to conditions in which learning is explicit. In the present study, we tested the hypothesis that SDC may also occur with implicit learning when learning benefits from the formation of contextual associations, a function associated with the hippocampus. Three versions of the serial-reaction-time task (SRTT) were examined, and SDC was assessed by comparing performance after 12 hr breaks that included or did not include sleep. SDC was observed in the Explicit condition. Two implicit conditions were compared. In the Implicit Noncontextual condition, participants performed a concurrent tone-counting task with the pitch of each tone selected at random, precluding cross-dimensional associations. In the Implicit Contextual condition, participants responded to the color of the cues while the spatial location of the cues followed a correlated sequence. Whereas learning was observed in both implicit conditions, SDC was restricted to the contextual condition. Given that the formation of contextual associations is dependent on the hippocampus, we suggest that SDC is a hippocampus-mediated process. PMID- 16713958 TI - The coiled-coil protein shrub controls neuronal morphogenesis in Drosophila. AB - The diversity of neuronal cells, especially in the size and shape of their dendritic and axonal arborizations, is a striking feature of the mature nervous system. Dendritic branching is a complex process, and the underlying signaling mechanisms remain to be further defined at the mechanistic level. Here we report the identification of shrub mutations that increased dendritic branching. Single cell clones of shrub mutant dendritic arborization (DA) sensory neurons in Drosophila larvae showed ectopic dendritic and axonal branching, indicating a cell-autonomous function for shrub in neuronal morphogenesis. shrub encodes an evolutionarily conserved coiled-coil protein homologous to the yeast protein Snf7, a key component in the ESCRT-III (endosomal sorting complex required for transport) complex that is involved in the formation of endosomal compartments known as multivesicular bodies (MVBs). We found that mouse orthologs could substitute for Shrub in mutant Drosophila embryos and that loss of Shrub function caused abnormal distribution of several early or late endosomal markers in DA sensory neurons. Our findings demonstrate that the novel coiled-coil protein Shrub functions in the endosomal pathway and plays an essential role in neuronal morphogenesis. PMID- 16713959 TI - Early segregation of germ and somatic lineages during gonadal regeneration in the annelid Enchytraeus japonensis. AB - Although regeneration studies are useful for understanding how organs renew, little information is available about regeneration of reproductive organs and germ cells. We here describe the behavior of germ-cell precursors during regeneration of the oligochaete annelid worm Enchytraeus japonensis, which has the remarkable feature of undergoing asexual (by fission) and sexual reproduction . We first found that the gonad can regenerate from any body fragment yielded by fission during asexual reproduction. We then examined behavior of germ-cell lineage during this regenerative process, by using a homolog of the Piwi gene (Ej piwi) as a marker. We found that in asexually growing animals, specialized cells expressing Ej-piwi are distributed widely in the body as single cells. These cells seem to serve as a reservoir of germ-cell precursors because during asexual propagation these cells migrate into the regenerating tissue, where they ultimately settle in the prospective gonads, and give rise to germ cells upon sexualization. These cells are distinct from the neoblasts, thought to be stem cells in other animals. This is the first report to directly show that the germ and somatic lineages are segregated in asexually growing animals and behave differently during regeneration. PMID- 16713960 TI - Transcriptional pulsing of a developmental gene. AB - It has not been possible to view the transcriptional activity of a single gene within a living eukaryotic cell. It is therefore unclear how long and how frequently a gene is actively transcribed, how this is modulated during differentiation, and how transcriptional events are dynamically coordinated in cell populations. By means of an in vivo RNA detection technique , we have directly visualized transcription of an endogenous developmental gene. We found discrete "pulses" of gene activity that turn on and off at irregular intervals. Surprisingly, the length and height of these pulses were consistent throughout development. However, there was strong developmental variation in the proportion of cells recruited to the expressing pool. Cells were more likely to re-express than to initiate new expression, indicating that we directly observe a transcriptional memory. In addition, we used a clustering algorithm to reveal synchronous transcription initiation in neighboring cells. This study represents the first direct visualization of transcriptional pulsing in eukaryotes. Discontinuity of transcription may allow greater flexibility in the gene expression decisions of a cell. PMID- 16713961 TI - Modeling clinically heterogeneous presenilin mutations with transgenic Drosophila. AB - To assess the potential of Drosophila to analyze clinically graded aspects of human disease, we developed a transgenic fly model to characterize Presenilin (PS) gene mutations that cause early-onset familial Alzheimer's disease (FAD). FAD exhibits a wide range in severity defined by ages of onset from 24 to 65 years . PS FAD mutants have been analyzed in mammalian cell culture, but conflicting data emerged concerning correlations between age of onset and PS biochemical activity . Choosing from over 130 FAD mutations in Presenilin-1, we introduced 14 corresponding mutations at conserved residues in Drosophila Presenilin (Psn) and assessed their biological activity in transgenic flies by using genetic, molecular, and statistical methods. Psn FAD mutant activities were tightly linked to their age-of-onset values, providing evidence that disease severity in humans primarily reflects differences in PS mutant lesions rather than contributions from unlinked genetic or environmental modifiers. Our study establishes a precedent for using transgenic Drosophila to study clinical heterogeneity in human disease. PMID- 16713963 TI - Aging and innate immunity. AB - Adaptive immunity undergoes severe deterioration with age and represents the main problem in the elderly. However, evidence accumulated over the last decade supports the hypothesis that aging also has a profound impact on innate immunity, which in turn markedly impacts the health and longevity of older people. PMID- 16713964 TI - Aging of the immune system: how much can the adaptive immune system adapt? AB - The competency of the adaptive immune function decreases with age, primarily because of the decline in production of naive lymphocytes in the bone marrow and thymus as well as the expansion of incompetent memory lymphocytes. Here I discuss the recent progress on age-associated changes in lymphocytes and their effect on the adaptive immune system. PMID- 16713965 TI - T cell anergy: where it's LAT. AB - T cell receptor engagement activates selective signaling pathways in T lymphocytes under different conditions. In this issue of Immunity, demonstrate that anergic T cells are selectively defective in LAT activation. PMID- 16713966 TI - Upstream toward the "DRiP"-ing source of the MHC class I pathway. AB - MHC class I binding peptides are generated via cytosolic degradation of a previously undefined substrate. In this issue of Immunity, pre-degradation polypeptide intermediates bound to a cytosolic chaperone is isolated. PMID- 16713967 TI - Unraveling the pros and cons of interferon-gamma gene regulation. AB - Although transforming growth factor-beta (TGF-beta) inhibition of interferon gamma (IFN-gamma) expression has been known for some time, in this issue of Immunity, the crosstalk between proinflammatory cytokine and TGF-beta signaling that regulates IFN-gamma expression is detailed for the first time. PMID- 16713968 TI - CCR6(+) dendritic cells: the gut tactical-response unit. AB - How an immune response to pathogens is initiated in the gut is still an open question. In this issue of Immunity, it has been found that a distinct subset of CCR6-expressing dendritic cells are absolutely required to activate anti Salmonella-specific T cells. PMID- 16713969 TI - Parallel pathways of virus recognition. AB - Viruses trigger signaling pathways of innate immunity. In this issue, it is shown that the mitochondrial antiviral signaling protein is critical for intracellular detection signaling, but is dispensable for the activation of innate immunity via Toll-like receptors. PMID- 16713970 TI - Impaired activation and localization of LAT in anergic T cells as a consequence of a selective palmitoylation defect. AB - The molecular basis of T cell anergy is not completely understood. We show that in antigen-primed anergic murine CD4(+) T cells the linker for activation of T cells (LAT) is hypophosphorylated upon CD3/CD28 restimulation. Signaling events downstream of LAT (PLCgamma1 phosphorylation and p85 [PI3-K] association) were impaired, whereas upstream events (CD3zeta and ZAP-70 phosphorylation) remained intact. LAT recruitment to the immunological synapse and its localization in detergent-resistant membrane (DRM) fractions were defective in anergic T cells. These defects resulted from impaired palmitoylation of LAT and were selective since the DRM localization and palmitoylation of Fyn were intact. This LAT defect was independent of Cbl-b and did not reflect enhanced LAT degradation. These results identify LAT as the most upstream target of anergy induction; moreover, they suggest that regulation of the amount of LAT in the immunological synapse and DRM by posttranslational palmitoylation contributes to the induction of T cell anergy. PMID- 16713971 TI - Hsp90alpha chaperones large C-terminally extended proteolytic intermediates in the MHC class I antigen processing pathway. AB - Intracellular proteins are degraded in the antigen processing pathway to generate peptide-loaded MHC I complexes (pMHC I) for immune surveillance. The characteristics of the final pMHC I are clear but those of their precursors and their potential binding partners remain poorly defined. By using a unique method to biochemically detect preprocessed ovalbumin-derived antigenic peptides, we find that cells generate large, C-terminally extended proteolytic intermediates that are associated with the alpha isotype of hsp90 chaperone. Knockdown of hsp90alpha expression by siRNA resulted in the loss of these intermediates and decreased presentation of the final pMHC I on the cell surface. Generation of pMHC I was also inhibited by knockdown of the cochaperone CHIP that interacts with heat shock proteins, ubiquitinates their clients, and delivers them to the proteasome. Thus, hsp90alpha can serve as a chaperone for precursors of pMHC I at an early stage in the antigen processing pathway. PMID- 16713972 TI - Diacylglycerol and protein kinase D localization during T lymphocyte activation. AB - The serine kinase protein kinase D (PKD) has a cysteine-rich domain (CRD) that binds diacylglycerol (DAG) with high affinity. PKD is cytosolic in unstimulated T cells, but it rapidly polarizes to the immunological synapse in response to antigen/antigen presenting cells (APCs). PKD repositioning is determined by the accumulation of DAG at the immunological synapse and changes in DAG accessibility of the PKD-CRD. Unstimulated T cells are shown to have a uniform distribution of DAG at the plasma membrane, whereas after T cell activation, a gradient of DAG is created with a persistent focus of DAG at the center of the synapse. PKD is only transiently associated with the immune synapse, indicating a fine tuning of PKD responsiveness to DAG by additional regulatory mechanisms. These results reveal the immune synapse as a focal point for DAG and PKD as an immediate and dynamic DAG effector during T cell activation. PMID- 16713973 TI - Human immunodeficiency virus type-1 infection impairs the formation of the immunological synapse. AB - HIV-1-infected lymphocytes improperly respond to T cell antigen receptor (TCR) stimulation. To document this phenomenon, we studied the capacity of HIV-1 infected lymphocytes to form immunological synapses. We show here that HIV-1 infected T cells poorly conjugated with antigen-presenting cells, and when they formed conjugates, the synapses were abnormal. TCR and Lck accumulated in the recycling endosomal compartment, and their clustering at the synapse was severely reduced. These phenomena were, to a large extent, caused by Nef, a viral protein affecting intracellular trafficking and signaling pathways. Concomitantly, in HIV infected cells, tyrosine phosphorylation at the synapse and the patterns of tyrosine phosphorylated proteins were disturbed in a Nef-dependent manner. These findings underscore the importance of Lck and TCR endosomal trafficking in synapse formation and early T cell signaling. Alteration of endocytic and signaling networks at the immunological synapse likely impacts the function and fate of HIV-1-infected cells. PMID- 16713974 TI - IFN-gamma suppresses IL-10 production and synergizes with TLR2 by regulating GSK3 and CREB/AP-1 proteins. AB - The control of IL-10 production and mechanisms that mediate synergy between IFN gamma and TLR ligands are not well understood. We report that IFN-gamma augments induction of TNFalpha by TLR ligands, immune complexes, and zymosan by suppressing IL-10 production and thereby interrupting Stat3-mediated feedback inhibition. IFN-gamma altered TLR2-induced signal transduction by increasing GSK3 activity and suppressing MAPK activation, leading to diminished IL-10 production. Inhibition of GSK3 or ablation of the GSK3beta gene ameliorated TLR2-induced peritonitis and arthritis. IFN-gamma suppressed the activity of CREB and AP-1, transcription factors that induce IL-10 expression and are regulated in part by MAPKs and GSK3. These results yield insight into mechanisms by which IFN-gamma regulates IL-10 production and TLR2-mediated inflammatory responses and identify inhibition of CREB and AP-1 as part of the macrophage response to IFN-gamma. GSK3 and CREB/AP-1 are key players in integrating IFN-gamma and TLR2 responses in innate immunity and inflammation. PMID- 16713975 TI - Pro- and antiinflammatory cytokine signaling: reciprocal antagonism regulates interferon-gamma production by human natural killer cells. AB - Activated monocytes produce proinflammatory cytokines (monokines) such as interleukin (IL)-12, IL-15, and IL-18 for induction of interferon-gamma (IFN gamma) by natural killer (NK) cells. NK cells provide the antiinflammatory cytokine transforming growth factor (TGF)-beta, an autocrine/negative regulator of IFN-gamma. The ability of one signaling pathway to prevail over the other is likely important in controlling IFN-gamma for the purposes of infection and autoimmunity, but the molecular mechanism(s) of how this counterregulation occurs is unknown. Here we show that in isolated human NK cells, proinflammatory monokines antagonize antiinflammatory TGF-beta signaling by downregulating the expression of the TGF-beta type II receptor, and its signaling intermediates SMAD2 and SMAD3. In contrast, TGF-beta utilizes SMAD2, SMAD3, and SMAD4 to suppress IFN-gamma and T-BET, a positive regulator of IFN-gamma. Indeed, activated NK cells from Smad3(-/-) mice produce more IFN-gamma in vivo than NK cells from wild-type mice. Collectively, our data suggest that pro- and antiinflammatory cytokine signaling reciprocally antagonize each other in an effort to prevail in the regulation of NK cell IFN-gamma production. PMID- 16713976 TI - Semaphorin 7A is a negative regulator of T cell responses. AB - Semaphorins play an essential role in axonal guidance, and emerging evidence points to diverse functions of several Semaphorin family members in the immune system. Semaphorin 7A (Sema7A) promotes axonal growth in the central nervous system. Here, we show that Sema7A also plays a critical role in negative regulation of T cell activation and function. T cells deficient in Sema7A exhibit enhanced homeostatic and antigen-induced proliferative response. Moreover, autoreactive Sema7A-deficient T cells mediate aggressive autoimmune disease. The deficiency in Sema7A leads to defective TCR downmodulation and T cell hyperresponsiveness. These results demonstrate an important role of Sema7A in limiting autoimmune responses and add to growing evidence of shared signaling pathways used by the immune and nervous systems. PMID- 16713977 TI - Carbon monoxide orchestrates a protective response through PPARgamma. AB - Carbon monoxide (CO) suppresses proinflammatory responses in macrophages reacting to LPS. We hypothesize that CO acts by inducing a molecule(s) that suppresses the inflammatory response to subsequent stress. Exposure of macrophages to CO alone in vitro produced a brief burst of mitochondrial-derived ROS, which led to expression of PPARgamma. PPARgamma expression proved essential for mediating the anti-inflammatory effects of CO. Blocking the CO-mediated increase in ROS generation prevented PPARgamma induction, and blocking PPARgamma prevented CO's anti-inflammatory effects. In a model of acute lung injury in mice, CO blocked expression of Egr-1, a central mediator of inflammation, and decreased tissue damage; inhibition of PPARgamma abrogated both effects. These data identify the mitochondrial oxidases as an (perhaps the) initial cellular target of CO and demonstrate that CO upregulates expression of PPARgamma via the mitochondria, which assures that a subsequent stress stimulus will lead to a cytoprotective as opposed to a proinflammatory phenotype. PMID- 16713978 TI - Crucial role of MLL for the maintenance of memory T helper type 2 cell responses. AB - The Mixed-Lineage Leukemia (MLL) gene, a mammalian homolog of the Drosophila trithorax, is implicated in regulating the maintenance of Hox gene expression and hematopoiesis. The physiological functions of MLL in the immune system remain largely unknown. Although MLL(+/-) CD4 T cells differentiate normally into antigen-specific effector Th1/Th2 cells in vitro, the ability of memory Th2 cells to produce Th2 cytokines was selectively reduced. Furthermore, histone modifications at the Th2 cytokine gene loci were not properly maintained in MLL(+/-) memory Th2 cells. The reduced expression of MLL in memory Th2 cells resulted in decreased GATA3 expression accompanied with impaired GATA3 locus histone modifications. The direct association of MLL with the GATA3 locus and the Th2 cytokine gene loci was demonstrated. Memory Th2 cell-dependent allergic airway inflammation was decreased in MLL(+/-) Th2 cell-transferred mice. Thus, a crucial role for MLL in the maintenance of memory Th2 cell function is indicated. PMID- 16713979 TI - CCR6-mediated dendritic cell activation of pathogen-specific T cells in Peyer's patches. AB - T cell activation by dendritic cells (DCs) is critical to the initiation of adaptive immune responses and protection against pathogens. Here, we demonstrate that a specialized DC subset in Peyer's patches (PPs) mediates the rapid activation of pathogen specific T cells. This DC subset is characterized by the expression of the chemokine receptor CCR6 and is found only in PPs. CCR6(+) DCs were recruited into the dome regions of PPs upon invasion of the follicle associated epithelium (FAE) by an enteric pathogen and were responsible for the rapid local activation of pathogen-specific T cells. CCR6-deficient DCs were unable to respond to bacterial invasion of PPs and failed to initiate T cell activation, resulting in reduced defense against oral infection. Thus, CCR6 dependent regulation of DCs is responsible for localized T cell dependent defense against entero-invasive pathogens. PMID- 16713980 TI - The specific and essential role of MAVS in antiviral innate immune responses. AB - The mitochondrial antiviral signaling protein (MAVS) mediates the activation of NFkappaB and IRFs and the induction of interferons in response to viral infection. In vitro studies have also suggested that MAVS is required for interferon induction by cytosolic DNA, but the in vivo evidence is lacking. By generating MAVS-deficient mice, here we show that loss of MAVS abolished viral induction of interferons and prevented the activation of NFkappaB and IRF3 in multiple cell types, except plasmacytoid dendritic cells (pDCs). However, MAVS was not required for interferon induction by cytosolic DNA or by Listeria monocytogenes. Mice lacking MAVS were viable and fertile, but they failed to induce interferons in response to poly(I:C) stimulation and were severely compromised in immune defense against viral infection. These results provide the in vivo evidence that the cytosolic viral signaling pathway through MAVS is specifically required for innate immune responses against viral infection. PMID- 16713982 TI - Fragility and crash over-representation among older drivers in Western Australia. AB - This study assessed age-related and gender differences in the relative contribution of fragility and crash over-representation to serious injuries per crash-involved driver in Western Australia. Police-reported crashes for the period 1998-2003 were extracted from the Western Australian Road Injury Database. For each passenger vehicle driver age and gender group, serious injuries per crash-involved driver and driver involvements in crashes per 100 million vehicle kilometre travelled (VKT) were calculated as the respective measure of fragility and crash over-representation. Results from the decomposition method of analysis showed that older drivers over the age of 70 sustained serious injury rates more than twice as high as those of the 30-59-year-old drivers. Fragility increased with age, contributing between 47% and 95% for drivers above 65 years, but crash over-representation was the dominant factor for male drivers above 80 years. In contrast, fragility contributed little to the excess injury risk of younger drivers under the age of 30. The importance of fragility as a contributing factor to the inflated serious injury risk per vehicle-kilometre travelled for older drivers suggested that road safety initiatives should be directed towards the protection of vehicle occupants as well as screening for their driving ability. PMID- 16713983 TI - Kinetic assay for measurement of enzyme concentration in situ. PMID- 16713981 TI - Skin-derived dendritic cells induce potent CD8(+) T cell immunity in recombinant lentivector-mediated genetic immunization. AB - The skin contains readily accessible dendritic cells (DCs) with potent antigen presentation function and functional plasticity enabling the integration of antigen specificity with environmentally responsive immune control. Recent studies challenge the established paradigm of cutaneous immune function by suggesting that lymph node-resident DCs, rather than skin-derived DCs (sDCs), are responsible for eliciting T cell immunity against cutaneous pathogens including viral vectors. We show that cutaneous delivery of lentivirus results in direct transfection of sDCs and potent and prolonged antigen presentation. Further, sDCs are the predominant antigen-presenting cells for the induction of potent and durable CD8(+) T cell immunity. These results support the classical paradigm of cutaneous immune function and suggest that antigen presentation by sDCs contributes to the high potency of lentivector-mediated genetic immunization. PMID- 16713984 TI - One-step biotinylation procedure for carbohydrates to study carbohydrate-protein interactions. AB - Protein-carbohydrate interactions play crucial roles in numerous biological processes. To study these interactions, we developed a simple and fast procedure for the biotinylation of carbohydrates based on reductive amination. The method allows complete and stable biotinylation of small quantities of oligosaccharides and includes a rapid and simple procedure to remove excess labeling reagent. After biotinylation, the structural and biological integrity of the glycans was intact as determined by HPLC, mass spectrometry, and a plant lectin assay. By using the human C-type lectin DC-SIGN (dendritic cell-specific ICAM-3-grabbing nonintegrin), we demonstrate that the biotinylated glycans can be used in a glycan array to determine binding specificities of lectins. Moreover, we show that fluorescent beads coated with selected biotinylated glycans bind to DC-SIGN expressing dendritic cells in vitro. Finally, by using biotinylated high-mannose N-glycans, we could visualize DC-SIGN-expressing cells in lymph node tissue. The availability of easy biotinylation methods for oligosaccharides such as those described here greatly facilitates the functional analysis of lectins. In addition, the biotinylated glycans will be great tools for investigating functional lectin receptors in situ. PMID- 16713985 TI - Perfusion-culture-based secreted bioluminescence reporter assay in living cells. AB - Bioluminescence reporter proteins have been widely used in the development of tools for monitoring biological events in living cells. In this study, we describe the development of a reporter system with secreted Cypridina noctiluca luciferase (CLuc) for a pharmacological assay that is based on targeted promoter activity. A model cell line was established with Rat-1 fibroblasts expressing CLuc driven by the promoter of a circadian clock gene, Bmal1. To accurately assay for temporally secreted CLuc activity, a perfusion culture in which the promoter activity was sequentially monitored by the reporter activity in the perfusate was adopted. By pulsing with dexamethasone (DEX), a glucocorticoid (GC) analog, the profile of the reporter activity successfully showed diurnal fluctuation, which is a canonical expression pattern of the Bmal1 gene. Trial studies illustrated that the DEX-pulsed circadian oscillation was reasonably attenuated by RU486, a GC receptor antagonist. Moreover, SP600125, a c-Jun N-terminal kinase inhibitor, caused phase shifting of the rhythmicity. We conclude that the CLuc reporter assay in combination with perfusion culture is a suitable pharmacological tool for drug discovery. PMID- 16713986 TI - Characterization of a laboratory model of computer mouse use - applications for studying risk factors for musculoskeletal disorders. AB - In the present study, we assessed the wrist kinetics (range of motion, mean position, velocity and mean power frequency in radial/ulnar deviation, flexion/extension, and pronation/supination) associated with performing a mouse operated computerized task involving painting rectangles on a computer screen. Furthermore, we evaluated the effects of the painting task on subjective perception of fatigue and wrist position sense. The results showed that the painting task required constrained wrist movements, and repetitive movements of about the same magnitude as those performed in mouse-operated design tasks. In addition, the painting task induced a perception of muscle fatigue in the upper extremity (Borg CR-scale: 3.5, p<0.001) and caused a reduction in the position sense accuracy of the wrist (error before: 4.6 degrees , error after: 5.6 degrees , p<0.05). This standardized painting task appears suitable for studying relevant risk factors, and therefore it offers a potential for investigating the pathophysiological mechanisms behind musculoskeletal disorders related to computer mouse use. PMID- 16713987 TI - Purification and characterization of hatching enzyme from shrimp Penaeus chinensis. AB - By using Penaeus chorion as a specific substrate, the hatching enzyme (HE) from Penaeus chinensis was purified by gel-filtration and ion-exchange chromatography, and characterized in terms of its molecular weight and enzymatic properties in this study. It was found that the molecular weight of Penaeus HE is about 43.0 kDa in SDS-PAGE. The Penaeus HE had obvious choriolytic activity, which was optimal at pH 6.0 and temperature of 40 degrees C, respectively. The Km value of the HE for casein was 7.47 mg ml(-1). The HE activity was almost completely inhibited by SBTI, p-APMSF, bestatin, and NEM, greatly inhibited by ovomucoid, TLCK, IAM, chymostatin, and PMSF, and slightly inhibited by pepstatin A, TPCK, LBTI, and leupeptin. These results indicate that the HE is most probably a trypsin-type serine protease. Besides of these, the HE was extremely sensitive to EDTA, Zn2+, Ca2+, Mg2+, and Cu2+. Combined with the results that the EDTA pretreated HE activity could be perfectly recovered by Zn2+, it is indicated that shrimp HE is most probably a kind of Zn-metalloprotease. PMID- 16713988 TI - On the contribution of the positively charged headgroup of choline to substrate binding and catalysis in the reaction catalyzed by choline oxidase. AB - Recent kinetic studies established that the positive charge on the trimethylammonium group of choline plays an important role in substrate binding and specificity in the reaction catalyzed by choline oxidase. In the present study, pH and solvent viscosity effects with the isosteric analogue of choline 3,3-dimethyl-butan-1-ol have been used to further dissect the contribution of the substrate positive charge to substrate binding and catalysis in the reaction catalyzed by choline oxidase. Both the kcat and kcat/Km values with 3,3-dimethyl butan-1-ol increased to limiting values that were approximately 3- and approximately 400-times lower than those observed with choline, defining pKa values that were similar to the thermodynamic pKa value of approximately 7.5 previously determined. No effects of increased solvent viscosity were observed on the kcat and kcat/Km values with the substrate analogue at pH 8, suggesting that the chemical step of substrate oxidation is fully rate-limiting for the overall turnover and the reductive half-reaction in which the alcohol substrate is oxidized to the aldehyde. The kcat/Km value for oxygen determined with the substrate analogue was pH-independent in the pH range from 6 to 10, with an average value that was approximately 75-times lower than that previously determined with choline as substrate. These data are consistent with the positive charge headgroup of choline playing important roles for substrate binding and flavin oxidation, with minimal contribution to substrate oxidation. PMID- 16713989 TI - Preparation and characterization of a stratum corneum substitute for in vitro percutaneous penetration studies. AB - The intercellular stratum corneum (SC) lipids form the main barrier for diffusion of substances through the skin. A porous substrate covered with synthetic SC lipids would be an attractive model to study percutaneous penetration, hereby replacing native human SC. Prerequisite is that this stratum corneum substitute (SCS) is prepared with a uniform lipid composition and layer thickness. Furthermore, the lipid organization and orientation should resemble that in SC. The objective of this study was to investigate the utility of an airbrush spraying device to prepare a SCS composed of cholesterol, ceramides and free fatty acids on a polycarbonate filter. The results demonstrate that a proper choice of solvent mixture and lipid concentration is crucial to achieve a uniform distribution of the applied lipids over the filter surface. A smooth and tightly packed lipid layer is only obtained when the equilibration conditions are appropriately chosen. The SCS possesses two crystalline lamellar phases with periodicities similar to those present in native SC. The orientation of these lamellae is mainly parallel to the surface of the polycarbonate filter, which resembles the orientation of the intercellular SC lipids. In conclusion, the airbrush technique enables generation of a homogeneous SCS, which ultimately may function as a predictive in vitro percutaneous penetration model. PMID- 16713990 TI - Comparison between low-level 50 Hz and 900 MHz electromagnetic stimulation on single channel ionic currents and on firing frequency in dorsal root ganglion isolated neurons. AB - Alteration of membrane surface charges represents one of the most interesting effects of the electromagnetic exposure on biological structures. Some evidence exists in the case of extremely low frequency whereas the same effect in the radiofrequency range has not been detected. Changes in transmembrane voltages are probably responsible for the mobilization of intracellular calcium described in some previous studies but not confirmed in others. These controversial results may be due to the cell type under examination and/or to the permeability properties of the membranes. According to such a hypothesis, calcium oscillations would be a secondary effect due to the induced change in the membrane voltage and thus dependent on the characteristics of ionic channels present in a particular preparation. Calcium increases could suggest more than one mechanism to explain the biological effects of exposure due to the fact that all the cellular pathways using calcium ions as a second messenger could be, in theory, disturbed by the electromagnetic field exposure. In the present work, we investigate the early phase of the signal transmission in the peripheral nervous system. We present evidence that the firing rate of rat sensory neurons can be modified by 50/60 Hz magnetic field but not by low level 900 MHz fields. The action of the 50/60 Hz magnetic field is biphasic. At first, the number of action potentials increases in time. Following this early phase, the firing rate decreases more rapidly than in control conditions. The explanation can be found at the single-channel level. Dynamic action current recordings in dorsal root ganglion neurons acutely exposed to the electromagnetic field show increased functionality of calcium channels. In parallel, a calcium-activated potassium channel is able to increase its mean open time. PMID- 16713991 TI - A randomized controlled trial on the long-term effects of a 1-month behavioral weight control program assisted by computer tailored advice. AB - OBJECTIVES: To examine the long-term effects of a new behavioral weight control program (Kenkou-tatsujin, KT program). The program consisted of twice-interactive letter communications including computer-tailored personal advice on treatment needs and behavioral modification. DESIGN: A randomized controlled trial comparing Group KM: KT program with 6-month weight and targeted behavior's self monitoring, Group K: KT program only, Group BM: an untailored self-help booklet with 7-month self-monitoring of weight and walking, and Group B: the self-help booklet only. PARTICIPANTS: Two hundreds and five overweight Japanese females were recruited via a local newspaper. MEASUREMENTS: Weight loss (body weight, BMI, reduction quotient, etc.) and behavioral changes (daily eating, exercise and sleeping habits). FINDINGS: A significant weight loss was observed in all groups. At 1 month, Groups KM and K were superior, but at 7 months, the mean weight loss was significantly more in Group KM than the other 3 groups. At 7 months, 8 dietary habits and 4 physical activities were improved in all subjects. Habitual improvement was related to the weight loss in Groups KM and K at 1 month. PMID- 16713992 TI - Immunosuppressant triptolide inhibits dendritic cell-mediated chemoattraction of neutrophils and T cells through inhibiting Stat3 phosphorylation and NF-kappaB activation. AB - Triptolide, an active component purified from the medicinal plant Tripterygium wilfordii Hook F., is potent in anti-inflammation and immunosuppression. Dendritic cells (DC), one of important targets of immunosuppressants, play crucial roles in linking the innate immunity and adaptive immunity. However, the effects of triptolide on DC have not been fully elucidated. Chemoattraction of neutrophils and T cells by DC may favor their interactions and initiation of immune response. Here we demonstrate that triptolide significantly impairs DC mediated chemoattraction of neutrophils and T cells both in vitro and in vivo by suppressing DC production of CC and CXC chemokines including MIP-1alpha, MIP 1beta, MCP-1, RANTES, TARC, and IP-10 in response to LPS. Furthermore, triptolide mediated inhibition of NF-kappaB activation, Stat3 phosphorylation and increase of SOCS1 expression in DC may be involved in the inhibitory effect of triptolide. Our study provides a novel mechanistic explanation for the anti-inflammatory and immunosuppressive activities of triptolide. PMID- 16713993 TI - SmPKC1, a new protein kinase C identified in the platyhelminth parasite Schistosoma mansoni. AB - Schistosoma mansoni signal transduction pathways are promising sources of target molecules for the development of novel control strategies against this platyhelminth parasite of humans. Members of the protein kinase C (PKC) family play key roles in such pathways activated by both receptor tyrosine kinases and other receptors, controlling a variety of physiological processes. Here, we report the cloning and molecular characterization of the first PKC identified in S. mansoni. Structural analysis indicated that SmPKC1 exhibits all the features typical of the conventional PKC subfamily. The gene structure was determined in silico and found to comprise a total of 15 exons and 14 introns. This structure is highly conserved; all intron positions are also present in the human PKCbeta gene and most of the exon sizes are identical. Using PCR on genomic DNA we were able to show that putative orthologues of SmPKC1 are present in 9 Schistosoma species. SmPKC1 expression is developmentally regulated with the highest level of transcripts in miracidia, whereas SmPKC1 protein expression is higher in the sporocyst. The localization of SmPKC1 on the sporocyst ridge cyton and in schistosomula acetabular glands suggests that the enzyme plays a role in signal transduction pathways associated with larval transformation. PMID- 16713994 TI - A targeted molecular dynamics study of WPD loop movement in PTP1B. AB - Targeted molecular dynamics was used to examine the mechanism of WPD loop closure in PTP1B, which is essential for the activity of the enzyme. Two important regions are identified: the R-loop (residues 113-118), which assists in substrate binding, and the S-loop (residues 198-209), which undergoes a conformational change that appears to be vital for the movement of the WPD loop. The S-loop is adjacent to the alpha3-helix, and its conformational change is coupled with a change of interactions between the alpha3- and alpha7-helices. This latter observation is of particular interest in connection with a novel class of allosteric inhibitors of PTP1B [Wiesmann et al., Nat. Struc. Mol. Biol. 11 (2004) 730-737]. These compounds prevent the closure of the WPD loop, forcing the enzyme to remain in a catalytically inactive conformation, by blocking the rearrangement of the alpha3-helix relative to the alpha7-helix. PMID- 16713995 TI - Identification of genetic networks involved in the cell injury accompanying endoplasmic reticulum stress induced by bisphenol A in testicular Sertoli cells. AB - To identify detailed mechanisms by which bisphenol A (BPA), an endocrine disrupting chemical, induces cell injury in mouse testicular Sertoli TTE3 cells, we performed genome-wide microarray and computational gene network analyses. BPA (200muM) significantly decreased cell viability and simultaneously induced an increase in mRNA levels of HSPA5 and DDIT3, endoplasmic reticulum (ER) stress marker genes. Of the 22,690 probe sets analyzed, BPA down-regulated 661 probe sets and up-regulated 604 probe sets by >2.0-fold. Hierarchical cluster analysis demonstrated nine gene clusters. In decreased gene clusters, two significant genetic networks were associated with cell growth and proliferation and the cell cycle. In increased gene clusters, two significant genetic networks including many basic-region leucine zipper transcription factors were associated with cell death and DNA replication, recombination, and repair. The present results will provide additional novel insights into the detailed molecular mechanisms of cell injury accompanying ER stress induced by BPA in Sertoli cells. PMID- 16713996 TI - Over-expression of Kv1.5 in rat cardiomyocytes extremely shortens the duration of the action potential and causes rapid excitation. AB - BACKGROUND: Genetically abnormal action potential duration (APD) can be a cause of arrhythmias that include long and short QT interval syndrome. PURPOSE: The aim of this study was to evaluate the arrhythmogenic effect of short QT syndrome induced by the over-expression of Kv1.5 in rat. METHODS: From Sprague-Dawley rats on fetal days 18-19, cardiomyocytes were excised and cultured with and without transfection with the Kv-1.5 gene using an adenovirus vector. The expression of Kv1.5 was proven by immunohistochemistry and Western blot analysis. In the culture dish and in the whole cells, the electrical activities were recorded using the whole-cell patch-clamp technique and the effects of 4-AP and verapamil were tested. RESULTS: After transfection with Kv1.5 for 12h, immunohistochemical staining and Western blot analysis were positive for Kv1.5 while they were negative in the control transfected with only Lac-Z. In the culture dish, the myocytes showed spontaneous beating at 115beats/min (bpm) just prior to the transfection with Kv1.5 and increased to 367bpm at 24h. The control myocytes showed stable beating rates during culturing. 4-AP at 200microM slowed down the rate and verapamil abolished the beating. In the whole cells, the maximal resting membrane potential was slightly depolarized and APD was extremely abbreviated both at 50% and 90% of repolarization compared with those of the control. Rapid spontaneous activities were found in a single myocyte with Kv1.5 transfection and 4-AP slowed down the frequency of the activities with a reversal of the shortened APD. CONCLUSION: The over-expression of Kv1.5 induced short APD and triggered activities in rat cardiomyocytes. This model can be used to study the arrhythmogenic substrate of short QT syndrome. PMID- 16713997 TI - Effect of L-arginine on asymmetric dimethylarginine (ADMA) or homocysteine accelerated endothelial cell aging. AB - We investigated here the effect of l-arginine on asymmetric dimethylarginine (ADMA) or homocysteine-accelerated endothelial aging. Endothelial cells were cultured in medium containing 70micromol/L arginine until fourteenth passage. ADMA, dl-homocysteine, and l-arginine were replaced every 48h starting at the fourth passage. ADMA or homocysteine inhibited significantly the population doublings (PD) and accelerated the process of aging. Co-incubation with l arginine enhanced PD, inhibited senescence associated beta-galactosidase activity, and increased telomerase activity. This effect was associated with an increase in NO synthesis and NO synthase protein expression. Furthermore, l arginine-induced NO formation was accompanied by a reduction in oxidative stress and an increase in protein expression and enzyme activity of heme oxygenase (HO) 1. The NO synthase inhibitor l-NAME completely abolished the effect of l-arginine on ADMA or homocysteine-accelerated aging. These findings demonstrate that l arginine prevents the onset of endothelial aging in ADMA or homocysteine-treated cells by increasing NO formation and consequently the induction of HO-1. This might provide a new strategy to delay ADMA or homocysteine-accelerated aging. PMID- 16713998 TI - Murine leukemia virus vector integration favors promoter regions and regional hot spots in a human T-cell line. AB - Genomic analysis of integration will be important in evaluating the safety of human gene therapy with retroviral vectors. Here, we investigated MLV vector integration sites in human T-cells, since they are amenable to gene transfer studies, and have been used therapeutically in clinical trials. We mapped 340 MLV vector integration sites in the infected human T-cell clones we established. The data showed that MLV preferred integration near the transcription start sites (+/ 5kb), near CpG islands (+/-1kb), and within the first intron of RefSeq genes. We also identified MLV integration hot spots that contained three or more integrations within a 100kb region. RT-PCR revealed that mRNA-levels of T-cell clones that contained MLV integrations near transcription start sites or introns were dysregulated compared to the uninfected cells. These studies help define the profile of MLV integration in T-cells and the risks associated with MLV-based gene therapy. PMID- 16713999 TI - Multipotential nestin and Isl-1 positive mesenchymal stem cells isolated from human pancreatic islets. AB - Mesenchymal cells in the developing pancreas express the neural stem cell marker nestin and the transcription factor islet-1 (Isl-1). Using defined culture conditions we isolated on a single cell basis nestin producing cells from human pancreatic islets. These cells were immortalized with lentiviral vectors coding for telomerase and mBmi. They are positive for Isl-1 and nestin and have the potential to adopt a pancreatic endocrine phenotype with expression of critical transcription factors including Ipf-1, Isl-1, Ngn-3, Pax4, Pax6, Nkx2.2, and Nkx6.1 as well as the islet hormones insulin, glucagon, and somatostatin. In addition, they can be differentiated into human albumin producing cells in vivo when grafted into a SCID mouse liver. In accordance with a mesenchymal phenotype, the cells were also able to adopt adipocytic or osteocytic phenotypes in vitro. In conclusion, cultured pancreatic islets contain nestin and Isl-1 positive mesenchymal stem cells with multipotential developmental capacity. PMID- 16714001 TI - Characterization of HCoV-229E fusion core: implications for structure basis of coronavirus membrane fusion. AB - Human coronavirus 229E (HCoV-229E), a member of group I coronaviruses, has been identified as one of the major viral agents causing respiratory tract diseases in humans for nearly 40 years. However, the detailed molecular mechanism of the membrane fusion mediated by the spike (S) protein of HCoV-229E remains elusive. Here, we report, for the first time, a rationally designed fusion core of HCoV 229E (HR1-SGGRGG-HR2), which was in vitro produced in GST prokaryotic expression system. Multiple lines of experimental data including gel-filtration, chemical cross-linking, and circular diagram (CD) demonstrated that the HCoV-229E fusion core possesses the typical properties of the trimer of coiled-coil heterodimer (six alpha-helix bundle). 3D structure modeling presents its most-likely structure, similar to those of coronaviruses that have been well-documented. Collectively, HCoV-229E S protein belongs to the type I fusion protein, which is characterized by the existence of two heptad-repeat regions (HR1 and HR2), furthermore, the available knowledge concerning HCoV-229E fusion core may make it possible to design small molecule or polypeptide drugs targeting the membrane fusion, a crucial step of HCoV-229E infection. PMID- 16714000 TI - Depression of MAD2 inhibits apoptosis of gastric cancer cells by upregulating Bcl 2 and interfering mitochondrion pathway. AB - Mitotic arrest deficient 2 (MAD2) is an essential component of the mitotic spindle checkpoint pathway. It was previously shown to be associated with drug resistance of tumor cells. To further explore the roles of MAD2 in responses of gastric cancer cells to chemotherapy drugs, we constructed the siRNA vectors of MAD2 and transfected them into gastric cancer SGC7901 cells to inhibit expression of MAD2. MTT assay showed that the downregulation of MAD2 increased the resistance of SGC7901 cells to spindle inhibitors and DNA damaging agents. The apoptosis rates of gastric cancer cells transfected with MAD2-siRNA were 10.7% and 10%, respectively, after treated by 1.0microg/ml VCR and cisplatin. In contrast, the apoptosis rates of SGC7901 and SGC7901/psilencer3.1 induced by VCR were 43.2%, 38.7%; and that induced by cispaltin were 34.1%, 31.4%. The ratio of Bcl-2 to Bax was much higher in the MAD2-siRNA transfectants compared with the SGC7901/psilencer. In SGC7901/psilencer, cytochrome c and cleaved caspase 3 protein levels increased along with the exposure time increased. However, these protein levels of SGC7901/MAD2-siRNA had no changes during the drug treatment. These results indicate that down regulation of MAD2 could promote the drug resistance of gastric cancer cells and inhibit anticancer drugs induced-apoptosis by upregulating Bcl-2 and interfering the mitochondrion apoptosis pathway. PMID- 16714002 TI - In vitro selection of zinc finger DNA-binding proteins through ribosome display. AB - DNA-binding proteins with sequence specificities have a variety of applications. To create novel functional DNA-binding proteins, in vivo selection methods have been developed. There are, however, crucial problems with such methods, e.g., limitation of library size and difficulty of expression of toxic proteins for the host cells. In order to overcome these problems, we developed a novel way to select DNA-binding proteins using an in vitro ribosome display technique. The three zinc finger DNA-binding protein libraries, based on a Zif268 containing randomized sequence in each finger, were prepared and transcribed to mRNA in vitro. The ternary ribosomal complexes, formed by mRNA, ribosome, and translated DNA-binding protein during translation in a rabbit reticulocyte in vitro translation system, were selected with biotinylated target DNA fragments bound to streptavidin magnetic beads. The extracted mRNAs from the selected complexes were amplified using reverse transcription PCR and then sequenced. This is the first report of the selection of DNA-binding proteins involving an in vitro ribosome display technique. PMID- 16714003 TI - Astrocyte reactivity influences the number of presynaptic terminals apposed to spinal motoneurons after axotomy. AB - Although synaptic plasticity is a widespread phenomenon, the underlying mechanisms leading to its occurrence are virtually unknown. In this sense, glial cells, especially astrocytes, may have a role in network changes of the nervous system, influencing the retraction of boutons as well as providing a proper perisynaptic environment, thereby affecting the replacement of inputs. Interestingly, the glial reaction does vary between strains of rats and mice. In this sense, we present evidence that C57BL/6J and A/J isogenic mice present different astrocyte reactivity after a peripheral lesion in vivo as well as in vitro, by analyzing primary cell cultures. Such a difference in the glial reaction has a direct influence on in vivo number of pre-synaptic terminals and on in vitro synaptogenesis. PMID- 16714004 TI - ERPs dissociate the effects of switching task sets and task cues. AB - Recent studies have suggested that reaction time (RT) costs associated with switching tasks do not reflect an endogenous control process of task set reconfiguration [Logan, G. D., Bundesen, C., 2003. Clever Homunculus: Is There an Endogenous Act of Control in the Explicit Task-Cuing Procedure? J. Exp. Psychol. Hum. Percept. Perform. 29 (3), 575-599]. Participants randomly switched between two simple tasks. Task was cued 600 ms prior to stimulus presentation using either a color or shape cue. A significant RT task switch cost was found when controlling for either a repeat or switch in cue category. In comparison, a switch in cue category had no effect on RT, even when examined across a cumulative distribution. Electrophysiological data revealed early cue processing effects within the first 300 ms after cue onset. However, replicating previous findings, an increased parietal positivity was found for task switch relative to task repeat trials that emerged prior to stimulus onset. This suggests that task set reconfiguration processes are activated when switching between tasks and supports the usefulness of task-switching paradigms in investigating cognitive control processes. PMID- 16714005 TI - The relationship between homeostasis model assessment and cardiovascular risk factors in Iranian subjects with normal fasting glucose and normal glucose tolerance. AB - BACKGROUND: Insulin resistance is a complex problem which may not always correlate with all its cardiovascular risk factors in various populations. We investigated the relationship between homeostasis model assessment of insulin resistance (HOMA-IR) with cardiovascular risk factors in Iranian subjects with normal fasting glucose (NFG) and normal glucose tolerance (NGT). METHODS: Of the 605 subjects aged 25-79 y enrolled in this study, after the oral glucose tolerance test, 366 subjects aged 25-50 y and 135 aged >50 y were classified as NFG and NGT. Insulin resistance was estimated by the HOMA-IR. RESULTS: Women had higher values of body mass index (BMI), insulin and HOMA-IR than men in both age groups. The prevalence of insulin resistance, general and abdominal obesity, low HDL-C and physical inactivity was higher in women than men in the 2 age groups. Men had a higher prevalence of hypertension and hypertriglyceridemia in the group with age 25-50 y. The Pearson correlation controlled for age, BMI, waist circumference and physical activity showed that HOMA-IR had significant correlation with triglyceride and inversely associated with HDL-C in both sexes. In addition, the results of HOMA-IR quartiles demonstrated that the prevalence of hypertension, obesity, and low HDL-C was particular high in women with HOMA-IR >2.39. Multiple regression indicated that log HOMA-IR was independently predicted by BMI, triglyceride and HDL-C in men and BMI, HDL-C and waist-to-hip (WHR) ratio in women. CONCLUSIONS: HOMA-IR is associated with the features of metabolic syndrome with a sex difference in the degree and predictors of HOMA-IR and the frequency of cardiovascular risk factors. PMID- 16714006 TI - Anti-mitochondrial antibodies in patients with systemic lupus erythematosus: revealing the unforeseen. PMID- 16714007 TI - History of diagnostic enzymology: a review of significant investigations. AB - This review of the history of diagnostic enzymology includes numerous important publications which were published over 70 years ago by outstanding clinical enzymologists throughout the world. The review includes the clinical enzymology of cardiovascular diseases, skeletal muscle diseases, pulmonary diseases, effusions, neurologic diseases, hematologic diseases, pregnancy, pediatric diseases, cancer, pancreatitis, hepatic diseases, bone diseases and cancer of the prostate. The advent of clinical enzymology thus occurred many years ago. PMID- 16714008 TI - Detection of outliers and establishment of targets in external quality assessment programs. AB - BACKGROUND: The establishment of target values is important in external quality assessment (EQA) programs since most of the programs use the target as the assessment criterion. Results submitted to EQA programs usually are not Gaussian distributed due to the contamination with outliers. The traditional or robust statistical method can be chosen for the truncation of outliers. We investigated the two methods when setting targets for glucose in an EQA program. METHODS: The results of glucose were analyzed as an all-methods group and divided into four subgroups according to the analytical methods prior to testing of each data distribution. Based on the distribution, the traditional or robust statistical method was used to detect outliers. After removal of outliers, the data distributions were retested and if Gaussian distributions were obtained, the mean values were used as the target. RESULTS: Original data sets were not Gaussian distributed for all tested groups. Therefore, the robust statistical method was employed to detect outliers. After truncation of outliers, the data sets showed Gaussian distributions and the means were used to set target values. CONCLUSIONS: The targets of glucose were determined for all-methods and individual methods from the mean values following removal of outliers using the robust statistical method. This led to comparable targets among the tested groups. PMID- 16714009 TI - Real-time telomeric repeat amplification protocol using the duplex scorpion and two reverse primers system: the high sensitive and accurate method for quantification of telomerase activity. AB - BACKGROUND: Real-time quantitative TRAP assays for detection of telomerase activity have been recently developed to eliminate complex post-PCR procedures. However, all of them use the conventional TRAP assay that possesses an unpredictable cascade of events in PCR amplification caused by stagger annealing, which may affect the accuracy of quantitation. METHODS: A novel RTQ-TRAP method was developed by combining the duplex scorpion with modified TP-TRAP assay that has high fidelity PCR amplification of the telomerase product (DS/TP-TRAP). The synthesized oligonucleotide that represents telomerase products is used to set up a standard curve. RESULTS: The DS/TP-TRAP method gives the standard curve a dynamic range of 6 orders of magnitude (R(2)=0.9992). It optimizes PCR amplification efficiency and determines telomerase activity in a lower threshold cycle number (Ct value). The method is both accurate and reproducible to measure telomerase activity in human tumor cell lines, and linearity from 1 to 1000 cells could be obtained (R(2)=0.9926). For tumor samples, the results determined by the DS/TP-TRAP assay are comparable to the data obtained with the conventional TRAP method. CONCLUSIONS: The DS/TP-TRAP assay provides a high sensitive and accurate method for real-time quantitative detection of telomerase activity. It is thus a potential robust tool for application in cancer molecular diagnostics. PMID- 16714010 TI - Apolipoprotein E polymorphism in the Tunisian population: frequency and effect on lipid parameters. AB - OBJECTIVES: We determined the frequencies of apolipoprotein E (apo E) gene alleles and examined the association between apo E polymorphism and lipid parameters in a sample of the Tunisian population. DESIGN AND METHODS: Apo E polymorphism was investigated using PCR, and plasma lipid parameters were measured in 122 men and 111 women aged 35 to 87 years. RESULTS: The allele frequencies were epsilon2: 7.3%, epsilon3: 84.6%, and epsilon4: 8.1%. Apo E polymorphism was associated with significant differences (P<0.001) in total cholesterol, apo B and LDL cholesterol in both men and women. epsilon2 carriers had the lowest mean total cholesterol, apo B and LDL-C concentrations, and subjects with the epsilon4 allele had the highest levels. Triglycerides levels increased with the epsilon4 allele, but this did not reach statistical significance. These results remained unchanged after adjustment for age, body mass index, sex, hypertension, diabetes and smoking. However, in obese subjects (BMI>30 kg/m2), TG concentrations were significantly lower in individuals homozygous for the epsilon3 allele compared to those with the alleles epsilon2 or epsilon4. CONCLUSION: In this sample of the Tunisian population, the distribution of apo E gene alleles is similar to that observed in Southern European populations with low prevalence of the epsilon4 allele. Variations in the apo E gene play a role in determining plasma lipid levels. These data also suggest that effects of apo E alleles on lipids levels are partly dependent on environmental variables such as BMI. These findings highlight the importance of the gene/environment interaction on the deleterious effect of obesity on cardiovascular risk factors. PMID- 16714011 TI - Effect of Agkistroden blomhoffi (mamushi) on superoxide generation in human neutrophils. AB - BACKGROUND: The purpose of this study is to investigate the effect of Agkistroden blomhoffi (mamushi) aqueous extract on stimulus-induced superoxide generation in human neutrophils. METHODS: Superoxide generation was assayed by measuring the reduction of ferricytochrome C (cyt c) using a dual-beam spectrophotometer. The phosphorylation of neutrophil proteins and the translocation of cytosolic compounds to the cell membrane were detected using specific monoclonal antibodies. RESULTS: Mamushi aqueous extract markedly suppressed fMLP-induced superoxide generation in a concentration-dependent manner. fMLP-induced translocation of cytosolic compounds p47phox, p67phox and Rac to the cell membrane was suppressed in parallel to the suppression of superoxide generation. The fractions with MW below 10 kDa dose-dependently suppressed tyrosyl phosphorylation of 45.0, 77.6 and 99.5 kDa proteins in fMLP-treated human neutrophils; the inhibition of tyrosyl phosphorylation was in parallel to that of the fMLP-induced translocation of p47phox to the cell membrane and superoxide generation. CONCLUSIONS: Mamushi aqueous extract markedly suppressed fMLP-induced superoxide generation in human neutrophils and also suppressed the translocation of cytosolic compounds to the cell membrane and tyrosyl phosphorylation of proteins. The efficacious components are the fractions with MW below 10 kDa. O phosphoserylethanolamine, a new dipeptide isolated from mamushi, also suppressed fMLP-induced superoxide generation in human neutrophils in a dose-dependent manner. PMID- 16714012 TI - Effects of excitotoxic lesions of the medial prefrontal cortex on density of high affinity [125I-Tyr3]neurotensin binding sites within the ventral midbrain and striatum. AB - The present study was aimed at determining the extent to which excitotoxic lesions of the medial prefrontal cortex reduce neurotensin receptors within the striatum, the nucleus accumbens, the ventral tegmental area and the substantia nigra. The medial prefrontal cortex was unilaterally lesioned with ibotenic acid and 10 days later brain sections were processed for neurotensin receptor autoradiographic analysis using 0.1 nM [(125)I-Tyr3]neurotensin with, or without, levocabastine. Analysis revealed at least two sites, one levocabastine insensitive neurotensin NT(1) and one levocabastine-sensitive neurotensin NT(2) like. The proportion of the latter site was high within the caudal striatum, the nucleus accumbens and the medial prefrontal cortex. Lesions produced a 60% to 80% reduction in neurotensin NT(1) within the ipsilateral medial prefrontal cortex, but no change in the sub-cortical nuclei. An increase in neurotensin NT(2)-like receptors was found in ipsilateral dorso-caudal caudate. These results show that a significant amount of neurotensin NT(1) receptors are located on neurons within the medial prefrontal cortex but not on their efferent terminals. PMID- 16714013 TI - Effect of combined treatment with noradrenaline and serotonin reuptake inhibitors on conditioned freezing. AB - To clarify the therapeutic interaction between serotonin and noradrenaline reuptake inhibition on fear, this study examined the acute and subchronic effects of combined treatment with the selective serotonin reuptake inhibitor citalopram and the selective noradrenaline reuptake inhibitor reboxetine on the expression of conditioned fear (re-exposure to an environment paired previously with inescapable electric footshocks). After fear conditioning was achieved with footshocks, the drugs were administered to rats and freezing behavior, as an index of fear, was observed in the shock chamber. Acute and subchronic treatment with citalopram was reproducibly anxiolytic against conditioned freezing. Acute reboxetine worsened conditioned freezing and reversed the acute anxiolytic effects of citalopram, but this anxiogenic effect of noradrenaline reuptake inhibition was not observed after subchronic treatment. These results suggest that adding noradrenaline reuptake inhibitors to serotonin reuptake inhibitors adversely affects fear, at least with acute treatment. PMID- 16714014 TI - Cannabinoid signalling in TNF-alpha induced IL-8 release. AB - The molecular events mediating the immunomodulatory properties of cannabinoids have remained largely unresolved. We have therefore investigated the molecular mechanism(s) through which R-(+)-[2,3-dihydro-5-methyl-3-[(morpholinyl)methyl] pyrrolo[1,2,3-de]-1,4-benzoxazinyl]-(1-napthanlenyl) methanone (WIN55212-2) modulate production of interleukin-8 (IL-8) in HT-29 cells. Release of IL-8 induced by tumor necrosis factor-alpha (TNF-alpha) was determined by enzyme linked immunosorbent assay (ELISA). Changes in expression of inhibitory kappa B (IkappaB) were monitored by Western blotting and activation of nuclear factor kappa B (NF-kappaB) was determined in electrophoretic mobility shift assay (EMSAs). TNF-alpha induced release of IL-8 was inhibited by WIN55212-2 which also blocked the degradation of IkappaB-alpha and activation of NF-kappaB induced by TNF-alpha. These data provide strong evidence that WIN55212-2 may modulate IL-8 release by negatively regulating the signaling cascade leading to the activation of NF-kappaB. These findings highlight a potential mechanism for the immunomodulatory properties of cannabinoids and contribute towards acquiring a clear understanding of the role of cannabinoids in inflammation. PMID- 16714015 TI - Alpha 2A-adrenoceptors enhance the serotonergic effects of fluoxetine. AB - The ability of subtype-preferring alpha(2)-adrenoceptor antagonists to enhance the neurochemical effects of the antidepressant, fluoxetine, was evaluated by in vivo microdialysis. Combining the selective alpha(2A)-adrenoceptor antagonist, BRL-44408 (10 mg/kg, s.c.), with fluoxetine (30 mg/kg, s.c.) elevated the extracellular levels of serotonin (5-HT) and noradrenaline in the rat frontal cortex, an effect not observed following antidepressant treatment alone. In contrast, combining fluoxetine with the alpha(2B)- or alpha(2C)-adrenoceptor antagonists, imiloxan (10 mg/kg, s.c.) or rauwolscine (10 mg/kg, s.c.), respectively, did not similarly alter biogenic amine levels. Collectively, these results reveal a specific role for the alpha(2A)-adrenoceptor subtype in augmenting the neurochemical effects of antidepressants. PMID- 16714016 TI - Neural stem cells may be uniquely suited for combined gene therapy and cell replacement: Evidence from engraftment of Neurotrophin-3-expressing stem cells in hypoxic-ischemic brain injury. AB - Previously, we reported that, when clonal neural stem cells (NSCs) were transplanted into brains of postnatal mice subjected to unilateral hypoxic ischemic (HI) injury (optimally 3-7 days following infarction), donor-derived cells homed preferentially (from even distant locations) to and integrated extensively within the large ischemic areas that spanned the hemisphere. A subpopulation of NSCs and host cells, particularly in the penumbra, "shifted" their differentiation towards neurons and oligodendrocytes, the cell types typically damaged following asphyxia and least likely to regenerate spontaneously and in sufficient quantity in the "post-developmental" CNS. That no neurons and few oligodendrocytes were generated from the NSCs in intact postnatal cortex suggested that novel signals are transiently elaborated following HI to which NSCs might respond. The proportion of "replacement" neurons was approximately 5%. Neurotrophin-3 (NT-3) is known to play a role in inducing neuronal differentiation during development and perhaps following injury. We demonstrated that NSCs express functional TrkC receptors. Furthermore, the donor cells continued to express a foreign reporter transgene robustly within the damaged brain. Therefore, it appeared feasible that neuronal differentiation of exogenous NSCs (as well as endogenous progenitors) might be enhanced if donor NSCs were engineered prior to transplantation to (over)express a bioactive gene such as NT 3. A subclone of NSCs transduced with a retrovirus encoding NT-3 (yielding >90% neurons in vitro) was implanted into unilaterally asphyxiated postnatal day 7 mouse brain (emulating one of the common causes of cerebral palsy). The subclone expressed NT-3 efficiently in vivo. The proportion of NSC-derived neurons increased to approximately 20% in the infarction cavity and >80% in the penumbra. The neurons variously differentiated further into cholinergic, GABAergic, or glutamatergic subtypes, appropriate to the cortex. Donor-derived glia were rare, and astroglial scarring was blunted. NT-3 likely functioned not only on donor cells in an autocrine/paracrine fashion but also on host cells to enhance neuronal differentiation of both. Taken together, these observations suggest (1) the feasibility of taking a fundamental biological response to injury and augmenting it for repair purposes and (2) the potential use of migratory NSCs in some degenerative conditions for simultaneous combined gene therapy and cell replacement during the same procedure in the same recipient using the same cell (a unique property of cells with stem-like attributes). PMID- 16714017 TI - Human neurospheres derived from the fetal central nervous system are regionally and temporally specified but are not committed. AB - Proliferating single cells were isolated from various CNS regions (telencephalon, diencephalon, midbrain, cerebellum, pons and medulla, and spinal cord) of human fetal cadavers at 13 weeks of gestation and grown as neurospheres in long-term cultures. We investigated whether neural stem cells (NSCs) or progenitors within spheres have specific regional or temporal characteristics with regard to growth, differentiation, and region-specific gene expression, and whether these molecular specifications are reversible. Regardless of regional origin, all of the neurospheres were found to contain cells of different subtypes, which suggests that multipotent NSCs, progenitors or radial glial cells co-exist with restricted neuronal or glial progenitors within the neurospheres. Neurospheres from the forebrain grew faster and gave rise to significantly more neurons than did those from either the midbrain or hindbrain, and regional differences in neuronal differentiation appeared to be sustained during long-term passage of neurospheres in culture. There was also a trend towards a reduction in neuronal emergence from the respective neurospheres over time in culture, although the percentages of neurons generated from cerebellum-derived neurospheres increased dramatically. These results suggest that differences in neuronal differentiation for the various neurospheres are spatially and temporally determined. In addition, the properties of glial fibrillary acidic protein (GFAP)-, glutamate-, and gamma aminobutyric acid (GABA)-expressing cells derived from neurospheres of the respective CNS regions appear to be regionally and temporally different. Isolated human neurospheres from different CNS compartments expressed distinctive molecular markers of regional identity and maintained these patterns of region specific gene expression during long-term passage in vitro. To determine the potential of human neurospheres for regional fate plasticity, single spheres from the respective regions were co-cultured with embryonic day 16.5 (E16.5 d) mouse brain slices. Specific cues from the developing mouse brain tissues induced the human neurospheres to express different marker genes of regional identity and to suppress the expression of their original marker genes. Thus, even the early regional identities of human neurospheres may not be irreversible and may be altered by local inductive cues. These findings have important implications for understanding the characteristics of growth, differentiation, and molecular specification of human neurospheres derived from the developing CNS, as well as the therapeutic potential for neural repair. PMID- 16714018 TI - A model for co-translational translocation: ribosome-regulated nascent polypeptide translocation at the protein-conducting channel. AB - The protein-conducting channel (PCC) must allow both the translocation of soluble polypeptide regions across, and the lateral partitioning of hydrophobic transmembrane helices (TMHs) into, the membrane. We have analyzed existing structures of ribosomes and ribosome-PCC complexes and observe conformational changes suggesting that the ribosome may sense and orient the nascent polypeptide and also facilitate conformational changes in the PCC, subsequently directing the nascent polypeptide into the appropriate PCC-mediated translocation mode. The PCC is predicted to be able to accommodate one central, consolidated channel or two segregated pores with different lipid accessibilities, which may enable the lipid mediated partitioning of a TMH from one pore, while the other, aqueous, pore allows translocation of a hydrophilic polypeptide segment. Our hypothesis suggests a plausible mechanism for the transitioning of the PCC between different configurations. PMID- 16714019 TI - Characterization of dimeric ATP synthase and cristae membrane ultrastructure from Saccharomyces and Polytomella mitochondria. AB - There is increasing evidence now that F(1)F(0) ATP synthase is arranged in dimers in the inner mitochondrial membrane of several organisms. The dimers are also considered to be the building blocks of oligomers. It was recently found that the monomers in beef and the alga Polytomella ATP synthase dimer make an angle of approximately 40 degrees and approximately 70 degrees, respectively. This arrangement is considered to induce a strong local bending of the membrane. To further understand the packing of dimers into oligomers we performed an electron microscopy analysis of ATP synthase dimers purified from Saccharomyces cerevisiae. Two types of dimers were found in which the angle between the monomers is either approximately 90 degrees or approximately 35 degrees. According to our interpretation, the wide-angle dimers (70-90 degrees) are "true dimers" whereas the small-angle dimers (35-40 degrees) rather are "pseudo dimers", which represent breakdown products of two adjacent true dimers in the oligomer. Ultrathin sectioning of intact Polytomella mitochondria indicates that the inner mitochondrial or cristae membrane is folded into lamellae and tubuli. Oligomers of ATP synthase can arrange in a helical fashion in tubular-shaped cristae membranes. These results strongly support the hypothesized role of ATP synthase oligomers in structural determination of the mitochondrial inner membrane. PMID- 16714020 TI - Differences in the regulation of microtubule stability by the pro-rich region variants of microtubule-associated protein 4. AB - We have recently reported a neural variant of microtubule-associated protein 4 with a short pro-rich region (MAP4-SP). Here, we show that the neural MAP4 has reduced microtubule-stabilizing activity, compared to the ubiquitous MAP4 with a long pro-rich region (MAP4-LP), both in vitro and in vivo. Fluorescence recovery after photobleaching analyses revealed that the interaction of MAP4-SP with the microtubules is very rapid, with a half-time of fluorescence recovery of 7 +/- 2.36 s, compared to 19.5 +/- 3.03 s in case of MAP4-LP. The dynamic interaction of MAP4-SP with microtubules in neural cells may contribute to the dynamic behaviors of extending neurites. PMID- 16714021 TI - Genomic and experimental evidence for a potential sexual cycle in the pathogenic thermal dimorphic fungus Penicillium marneffei. AB - All meiotic genes (except HOP1) and genes encoding putative pheromone processing enzymes, pheromone receptors and pheromone response pathways proteins in Aspergillus fumigatus and Aspergillus nidulans and a putative MAT-1 alpha box mating-type gene were present in the Penicillium marneffei genome. A putative MAT 2 high-mobility group mating-type gene was amplified from a MAT-1 alpha box mating-type gene-negative P. marneffei strain. Among 37 P. marneffei patient strains, MAT-1 alpha box and MAT-2 high-mobility group mating-type genes were present in 23 and 14 isolates, respectively. We speculate that P. marneffei can potentially be a heterothallic fungus that does not switch mating type. PMID- 16714022 TI - Properties of the prolyl oligopeptidase homologue from Pyrococcus furiosus. AB - Prolyl oligopeptidase (POP), the paradigm of a serine peptidase family, hydrolyses peptides, but not proteins. The thermophilic POP from Pyrococcus furiosus (Pfu) appeared to be an exception, since it hydrolysed large proteins. Here we demonstrate that the Pfu POP does not display appreciable activity against azocasein. The autolysis observed earlier was an artefact. We have also found that the pH-rate profile is different from that of the mammalian enzyme and the low pK(a) extracted from the curve represents the ionization of the catalytic histidine. We conclude that some oligopeptidases may be true endopeptidases, cleaving at disordered segments of proteins, but with very low efficacy. PMID- 16714023 TI - New linker proteins in phycobilisomes isolated from the cyanobacterium Gloeobacter violaceus PCC 7421. AB - Two new linker proteins were identified by peptide mass fingerprinting in phycobilisomes isolated from the cyanobacterium Gloeobacter violaceus PCC 7421. The proteins were products of glr1262 and glr2806. Three tandem phycocyanin linker motifs similar to CpcC were present in each. The glr1262 product most probably functions as a rod linker connecting phycoerythrin and phycocyanin, while the glr2806 product may function as a rod-core linker. We have designated these two proteins CpeG and CpcJ, respectively. The morphology of phycobilisomes in G. violaceus has been reported to be a bundle-like shape with six rods, consistent with the proposed functions of these linkers. PMID- 16714024 TI - Characterization and quantification of the hydrocarbons fraction of the subcutaneous fresh fat of Iberian pig by off-line combination of high performance liquid chromatography and gas chromatography. AB - Hydrocarbons in fresh subcutaneous fat of Iberian pig have been analyzed by GC-MS after fractionation of the unsaponifiable fraction with a new off-line combination of HPLC and GC method. The new method proposed improves the recovery and simultaneous quantification of terpenic hydrocarbons in comparison to the traditional LC method. When necessary and for identification purposes, selective ion monitoring (SIM) was used as acquisition mode in GC-MS. To determine the position of the double bonds in the unsaturated hydrocarbon chain the dimethyl disulfide derivatives (DMDS) were obtained. To elucidate the structure of the branched 1-alkenes the hydrocarbon fraction was submitted to hydrogenation. Thirty-five compounds have been identified, including n-alkanes, n-alkenes, branched (n-1,n-2-dimethyl-1-alkenes) and terpenic hydrocarbons, being the most abundant n-alkenes and n-alkanes of even chain of n-C12-n-C26. Besides the hydrocarbons already described in bibliography, a new diterpenic hydrocarbon, ent kaurene, have been identified for the first time. The compound reported as Neophytadiene by other authors, has been identified as a 20 atoms hydrocarbon with two double bonds, the 7,11,15-trimethyl-heptadeca-1,4-diene. PMID- 16714025 TI - Rapid screening of polychlorinated biphenyls in sediments using non-equilibrium solid-phase microextraction and fast gas chromatography with electron-capture detection. AB - A rapid method for the screening of polychlorinated biphenyls (PCBs) in sediment samples is described. Analytes are concentrated on a poly(dimethylsiloxane) (PDMS) solid-phase microextraction (SPME) fibre, separated using a narrow-bore gas chromatography column (10 m x 0.1mm I.D.) and determined with a micro electron-capture detector (micro-ECD). The narrow-bore column showed a similar separation efficiency to a conventional capillary one (30 m x 0.32 mm I.D.) and allowed a 3 times reduction in the analysis time. Moreover, thermal desorption of SPME fibres in the narrow-bore column did not produce any degradation in the quality of separations when compared to the split injection mode. The use of an oxidative sample treatment was mandatory to remove sulphur interferences in the chromatograms and increase the yield of PCBs during SPME for short sampling times. Under optimised conditions, a fraction of sediment (0.1-0.5 g) was mixed with 5 ml of a 0.2M potassium permanganate solution and 0.5 ml of sulphuric acid 6M in a 22 ml closed vessel. Sample treatment and headspace SPME of PCBs were performed simultaneously for 10 min at 100 degrees C. Globally, the whole analytical procedure takes about 20 min and presents quantification limits between 0.5 and 1.3 ng/g. Accuracy was evaluated by analysing a reference sediment sample with certified concentrations of several PCBs. PMID- 16714026 TI - Simplified gradient generator for micro- and nano-liquid chromatography. AB - We have developed a simple device to generate gradient elution profiles using split tubing array (STAR) units for micro- and nano-HPLC. This gradient device consists of a delivery pump, a splitter, tubes in parallel, and a relatively large-volume mixing chamber. In the mixing chamber, an initially filled water rich solvent is adjusted to an organic-rich solvent by delivery of appropriate components through the split paths, in which the flow rate and residence time are controlled by the sizes of the tubes employed. A program was developed to describe the output gradient profiles, and the predicted gradient profiles were highly consistent with the observed ones. A linear gradient generated by this STAR system was successfully applied to micro-HPLC systems for separation of digested peptides derived from serum albumin with sufficient reproducibility. Further miniaturization of STAR systems for nano-HPLC coupled with tandem mass spectrometry was accomplished to separate digested peptides from serum albumin with a reproducibility of retention times of better than 0.2%, and the obtained spectra from the well-separated chromatographic peaks allowed protein identification with high confidence by protein database searching. We believe that this simple and reproducible gradient system using an isocratic pump will be especially advantageous for nanoLC-MS, instead of flow-controlled gradient pumps. PMID- 16714028 TI - New cold-fiber headspace solid-phase microextraction device for quantitative extraction of polycyclic aromatic hydrocarbons in sediment. AB - A new automated headspace solid-phase microextraction (HS-SPME) sampling device was developed, with the capability of heating the sample matrix and simultaneously cooling the fiber coating. The device was evaluated for the quantitative extraction of polycyclic aromatic hydrocarbons (PAHs) from solid matrices. The proposed device improves the efficiency of the release of analytes from the matrix, facilitates the mass transfer into the headspace and significantly increases the partition coefficients of the analytes, by creating a temperature gap between the cold-fiber (CF) coating and the hot headspace. The reliability and applicability of previously reported cold-fiber devices are significantly enhanced by this improvement. In addition, it can be easily adopted for full automation of extraction, enrichment and introduction of different samples using commercially available autosampling devices. Sand samples spiked with PAHs were used as solid matrices and the effect of different experimental parameters were studied, including the extraction temperature, extraction time, moisture content, and the effect of sonication and modifier under optimal experimental conditions, linear calibration curves were obtained in the range of 0.0009-1000 ng/g, with regression coefficients higher than 0.99 and detection limits that ranged from 0.3 to 3 pg/g. Reproducible, precise and high throughput extraction, monitoring and quantification of PAHs were achieved with the automated cold-fiber headspace solid-phase microextraction (CF-HS-SPME) device coupled to GC-flame ionization detection. Determination of PAHs in certified reference sediments using the proposed approach exhibited acceptable agreement with the standard values. PMID- 16714027 TI - High-performance thin-layer chromatographic fingerprints of isoflavonoids for distinguishing between Radix Puerariae Lobate and Radix Puerariae Thomsonii. AB - The roots of Pueraria lobata (Wild.) Ohwi and Pueraria thomsonii Benth have been officially recorded in all editions of Chinese Pharmacopoeia under the same monograph 'Gegen' (Radix Puerariae, RP). However, in its 2005 edition, the two species were separated into both individual monographs, namely 'Gegen' (Radix Puerariae Lobatae, RPL) and 'Fenge' (Radix Puerariae Thomsonii, RPT), respectively, due to their obvious content discrepancy of puerarin, the major active constituent. In present paper, the fingerprint of high-performance thin layer chromatography (HPTLC) combining digital scanning profiling was developed to identify and distinguish the both species in detail. The unique properties of the HPTLC fingerprints were validated by analyzing ten batches of Pueraria lobata and P. thomsonii samples, respectively. The common pattern of the HPTLC images of the roots of Pueraria spp. and the respective different ratios of the chemical distribution can directly discern the two species. The corresponding digital scanning profiles provided an easy way for quantifiable comparison among the samples. Obvious difference in ingredient content and HPTLC patterns of the two species questioned their bio-equivalence and explained that recording both species separately in the current edition of Chinese Pharmacopoeia (2005 edition) is reasonable due to not only the content of major constituent, puerarin, but also the peak-to-peak distribution in the fingerprint and integration value of the total components. Furthermore, the HPTLC fingerprint is also suitable for rapid and simple authentication and comparison of the subtle difference among samples with identical plant resource but different geographic locations. PMID- 16714029 TI - Urinary bladder lesions in bovine enzootic haematuria. AB - In cattle, bracken fern chronic toxicity is characterized by the presence of multiple tumours in the bladder (bovine enzootic haematuria). From October 1999 to March 2003, 433 urinary bladders with macroscopical lesions were collected in the slaughterhouse of Sao Miguel Island (Azores, Portugal), an endemic area where Pteridium aquilinum infestation in pastures is high. Bladder lesions were divided into three main categories (inflammatory lesions, non-neoplastic epithelial abnormalities and tumours) and described in detail. In some cases, neoplastic growth was confined to a single site, but in most cases multiple tumours developed within the same bladder. Epithelial tumours alone were present in 51.2% of the affected bladders, mesenchymal tumours alone in 17.4%, and both epithelial and mesenchymal tumours in the remaining 31.4%. The large number of tumours examined (870) revealed new categories not yet included in other veterinary classification systems, namely, inverted papilloma, papillary neoplasm of apparent low malignant potential, and haemangioendothelioma. PMID- 16714030 TI - Effect of vitamin E nutritional supplementation on the pathological changes induced in the ileum of rabbits by experimental infection with enteropathogenic Escherichia coli. AB - A well-established rabbit model of enteropathogenic E. coli (EPEC) disease was used to examine whether vitamin E (VE) nutritional supplementation had an effect on the pathological changes induced in the bowel by EPEC. Quantitative methods were used to evaluate the influence of VE on bacterial colonization, intestinal mucosal architecture and inflammation, and intestinal epithelial proliferation and apoptosis. VE did not affect EPEC colonization and did not give significant protection against EPEC-induced changes and diarrhoea. Although VE had no effect on the EPEC-related increase of enterocyte apoptosis, it clearly contributed to an acceleration of epithelial cell proliferation in the ileal crypts. This finding may explain why ileal morphometry undertaken in this study showed that VE ameliorated somewhat the effects of EPEC on intestinal mucosal architecture. Quantitative studies on inflammatory cells in the intestinal mucosa revealed that VE nutritional supplementation resulted in an increased neutrophilic and mononuclear inflammatory cell response to EPEC infection, which did not contribute, however, to the clearance of infection. PMID- 16714031 TI - Accounting for children's orthographic learning while reading text: do children self-teach? AB - Share's "self-teaching" model proposes that readers acquire most knowledge about the orthographic structure of words incidentally while reading independently. In the current study, the self-teaching hypothesis was tested by simulating everyday reading through the use of real words, analyzing the effects of context, and considering the independent contributions of general cognitive ability, including rapid naming ability and prior orthographic knowledge. A total of 35 first graders read short story passages in English embedded with target words representative of words likely to be known orally but not orthographically. Words were manipulated for target word spelling and contextual support. According to the self-teaching model, words correctly decoded during reading should be correlated with subsequent orthographic learning. The results of this study confirmed this prediction. Self-teaching was evidenced through significantly higher proportions of correctly identified target words across context conditions. Regression analyses showed that individual differences were related to prior orthographic knowledge and predicted students' degree and quality of orthographic learning after controlling for general decoding ability. PMID- 16714032 TI - Taking the task seriously: reflections on measures of color acquisition. AB - The experiments described in the lead articles by Kowalski and Zimiles and by O'Hanlon and Roberson examine factors that lead to color term acquisition. These experiments touch on the debate regarding the relative contributions of language and concepts in word learning. In this reflection, we examine how conclusions concerning the debate depend deeply on the particular task presented to children, and we propose an alternative approach to studying color term acquisition. PMID- 16714033 TI - A new sensitive and specific enzyme-linked immunosorbent assay for IgD. AB - We have developed a new highly specific ELISA for IgD, and then used it to measure levels of circulating IgD in the serum of 480 un-selected patients from the East Anglia region of UK. The assay is both extremely sensitive and specific, with a minimum detected IgD concentration of 30 pg/ml and more than 10,000-fold specificity for IgD over all other human immunoglobulins. The assay shows linear dilution characteristics with both purified IgD and human serum, and spiking of purified IgD into either purified immunoglobulins or human serum shows c. 100% recovery. Furthermore, intra-assay and inter-assay coefficients of variation for repeated measurements of the same samples are below 10% and 15% respectively. Measurement of IgD levels on the un-selected patient population showed levels to range from <300 pg/ml to over 100 microg/ml, with a geometric mean of 8 microg/ml. The distribution is approximately normal after log transformation. Levels of circulating IgD were higher in men than in women. There was a significant negative correlation between levels of IgD and age in women, but not in men. Moreover, after adjustment for age and sex, there were statistically significantly higher levels of circulating IgD in male (but not female) smokers, compared to their non-smoking counterparts. These results highlight the care that needs to be taken to control for age, sex and cigarette smoking when examining levels of circulating IgD in future studies. PMID- 16714034 TI - Norepinephrine-induced changes in gene expression of phospholipase C in cardiomyocytes. AB - Previously, we have reported that norepinephrine (NE)-mediated cardiac hypertrophy may occur due to stimulation of alpha1-adrenoceptors and phospholipase C (PLC) activity. Since the signal transduction mechanisms involving PLC isozymes in cardiomyocytes are not well established, the present study was conducted to test the hypothesis that stimulation of cardiac PLC activity by NE increases the gene expression for PLC isozymes via a PKC and ERK 1/2-dependent pathway. For this purpose, mRNA levels for PLC beta1, beta3, gamma1, and delta1 isozymes were determined in isolated adult rat cardiomyocytes upon incubation in the absence and presence of NE. The NE-induced increases in PLC isozyme mRNA levels were not only attenuated by prazosin, an inhibitor of alpha1-adrenergic receptor, but also by U73122, an inhibitor of PLC activity. Alterations in NE-induced PLC gene expression by both prazosin and U73122 were associated with inhibition of PLC activity. The inhibition of NE-stimulated PLC gene expression by bisindolylmaleimide, a PKC inhibitor, and PD98059, an ERK1/2 inhibitor, indicated that PKC-MAPK signaling may be involved in this signal transduction pathway. The observed NE-induced changes in gene expression in the presence of different inhibitors were associated with corresponding changes in the protein content. Furthermore, significant increases in mRNA levels and protein contents for all PLC isozymes were found in cardiomyocytes treated with phorbol 12-myristate 13-acetate, a PKC activator. These data indicate that PLC isozymes may regulate their own gene expression through a PKC and ERK 1/2 dependent pathway in a cycle of events associated with the cardiomyocyte hypertrophic response. PMID- 16714035 TI - Thermoregulatory role of inducible nitric oxide synthase in lipopolysaccharide induced hypothermia. AB - We have tested the hypothesis that nitric oxide (NO) arising from inducible nitric oxide synthase (iNOS) plays a role in hypothermia during endotoxemia by regulating vasopressin (AVP) release. Wild-type (WT) and iNOS knockout mice (KO) were intraperitoneally injected with either saline or Escherichia coli lipopolysaccharide (LPS) 10.0 mg/kg in a final volume of 0.02 mL. Body temperature was measured continuously by biotelemetry during 24 h after injection. Three hours after LPS administration, we observed a significant drop in body temperature (hypothermic response) in WT mice, which remained until the seventh hour, returning then close to the basal level. In iNOS KO mice, we found a significant fall in body temperature after the fourth hour of LPS administration; however, the hypothermic response persisted until the end of the 24 h of the experiment. The pre-treatment with beta-mercapto-beta,beta cyclopentamethylenepropionyl(1), O-Et-Tyr2, Val4, Arg8-Vasopressin, an AVP V1 receptor antagonist (10 microg/kg) administered intraperitoneally, abolished the persistent hypothermia induced by LPS in iNOS KO mice, suggesting the regulation of iNOS under the vasopressin release in this experimental model. In conclusion, our data suggest that the iNOS isoform plays a role in LPS-induced hypothermia, apparently through the regulation of AVP release. PMID- 16714036 TI - Atrial natriuretic peptide-dependent modulation of hypoxia-induced pulmonary vascular remodeling. AB - Hypoxic stress upsets the balance in the normal relationships between mitogenic and growth inhibiting pathways in lung, resulting in pulmonary vascular remodeling characterized by hyperplasia of pulmonary arterial smooth muscle cells (PASMCs) and fibroblasts and enhanced deposition of extracellular matrix. Atrial natriuretic peptide (ANP) reduces pulmonary vascular resistance and attenuates hypoxia-induced pulmonary hypertension in vivo and PASMC proliferation and collagen synthesis in vitro. The current study utilized an ANP null mouse model (Nppa-/-) to test the hypothesis that ANP modulates the pulmonary vascular and alveolar remodeling response to normobaric hypoxic stress. Nine-10 wk old male ANP null (Nppa-/-) and wild type nontransgenic (NTG) mice were exposed to chronic hypoxia (10% O(2), 1 atm) or air for 6 wks. MEASUREMENT: pulmonary hypertension, right ventricular hypertrophy, and pulmonary arterial and alveolar remodeling were assessed. Hypoxia-induced pulmonary arterial hypertrophy and muscularization were significantly increased in Nppa-/- mice compared to NTG controls. Furthermore, the stimulatory effects of hypoxia on alveolar myofibroblast transformation (8.2 and 5.4 fold increases in Nppa-/- and NTG mice, respectively) and expression of extracellular matrix molecule (including osteopontin [OPN] and periostin [PN]) mRNA in whole lung were exaggerated in Nppa-/- mice compared to NTG controls. Combined with our previous finding that ANP signaling attenuates transforming growth factor (TGF)-beta-induced expression of OPN and PN in isolated PASMCs, the current study supports the hypothesis that endogenous ANP plays an important anti-fibrogenic role in the pulmonary vascular adaptation to chronic hypoxia. PMID- 16714037 TI - Retrieval and emotional processing of traumatic memories in posttraumatic stress disorder: peripheral and central correlates. AB - Posttraumatic stress disorder (PTSD) is thought to be characterized by dysfunctional memory processes, i.e., the automatic re-experiencing of the traumatic event and the inability to consciously recall facts about the traumatic event, as well as altered emotional processing of trauma-relevant cues. The present study examined the cerebral mechanisms underlying the cued recall of trauma-specific memories and the emotional processing of the presented cues in 16 PTSD patients, 15 trauma-exposed subjects without PTSD and 16 healthy controls. Subjects received questions about their specific trauma as well as other disastrous and neutral events while the electroencephalogram and heart rate were measured. The PTSD patients showed no impairment in trauma-specific declarative memory compared to non-PTSD subjects but had some deficits in general declarative memory as assessed by the Wechsler Memory Scale-Revised. Compared to healthy control subjects, PTSD patients displayed increased P300 and late positive complex amplitudes to trauma-specific questions, indicating enhanced emotional processing of these cues. In line with their behavioral performance, both trauma exposed groups showed decreased terminal contingent negative variation amplitudes to trauma-specific questions over frontal electrodes reflecting altered memory retrieval. Within-group comparisons revealed that only the PTSD group but not the other groups showed a differentiation between trauma-specific and neutral questions with respect to the LPC, tCNV and P300. Concordantly with previous studies, PTSD patients showed elevated resting heart rate compared to the healthy controls. These findings are discussed in the context of current models of the role of declarative memory in the development and maintenance of PTSD. PMID- 16714038 TI - Metabolite profiling and characterization of somaclonal variants in tea (Camellia spp.) for identifying productive and quality accession. AB - A study has been undertaken to characterize 15 field grown somaclonal variants derived from cotyledonary tissues of UPASI-10 using morphological, physiological and biochemical characters. Although variants were derived from UPASI-10, a very few variants possessed unique "Chinery" characters while others exhibited "Assam" characters. However, no variant showed identical morphological characters aligning with the parent. Somaclonal variants showed distinct variation in terms of photosynthetic carbon assimilation, stomatal conductance and diffusion resistance. Proline accumulation and water use efficiency showed marginal variations among the variants. SE 8 and SE 10 recorded higher values of membrane stability index denoting their tolerant nature against stress. Class interval analysis based on physiological parameters grouped these plants into three clusters. Three variants grouped under good category representing higher values of productivity index followed by five variants under moderate category. Green leaf constituents and quality profile of made tea produced with crop shoots of variants exhibited wide variation. Center point radar graph analysis of quality constituents grouped these plants into three clusters. Variants SE 2 and SE 13 were segregated distinctly representing their black tea characters. When considering both the quality and productivity indices, SE 3 and SE 7 fall under moderate category and in future these two variants may be subjected to further quality tests for commercial exploitation. PMID- 16714039 TI - Scedosporium apiospermum mycetoma with bone involvement successfully treated with voriconazole. AB - Treatment of Scedosporium apiospermum mycetoma usually requires limb amputation. A 49-year-old woman, from Ivory Coast, was diagnosed with Madura foot in 1995. She failed to respond to several treatments including itraconazole, fluconazole and co-trimoxazole, and refused limb amputation. In December 2002 she was admitted to hospital in France with a painful, swollen right leg and foot. She had no fever and C-reactive protein was 120 mg/l. Magnetic resonance imaging (MRI) confirmed the destruction of tarsus bones with a tibia extension. Voriconazole (400 mg/day) treatment was initiated in March 2003; a significant clinical improvement was observed within 4 months as confirmed by C-reactive protein (16 mg/l) and MRI. Voriconazole was maintained for 18 months with good tolerance. Cholestasis appeared after the first month and remained stable. In October 2004 voriconazole was discontinued due to side effects on the liver (alanine aminotransferase 17 times the normal level); MRI showed impressive regression of bone lesions. As of July 2005, the patient remains clinically well. Voriconazole appears to be a promising drug for the treatment of S. apiospermum mycetomas. PMID- 16714040 TI - Emergence of Salmonella enterica serotype Paratyphi A as a major cause of enteric fever in Kathmandu, Nepal. AB - We performed pulsed-field gel electrophoresis (XbaI) on 114 bloodstream isolates of Salmonella enterica serotype Paratyphi A and S. enterica serotype Typhi collected from febrile patients in Kathmandu, Nepal. Of the 56 S. Paratyphi A isolates, 51 (91%) were indistinguishable, which suggests the emergence of a single clone. In contrast, only 21 (36%) of the 58 S. Typhi isolates exhibited a common genotype, which is consistent with endemic disease from multiple sources. PMID- 16714042 TI - Coded excitation using biphase-coded pulse with mismatched filters for high frequency ultrasound imaging. AB - A scheme of using phase-coded excitation and mismatched filter compression for high-frequency ultrasound imaging is presented in this paper. Biphase-coded pulses were constructed to excite the transducer. Received signals were compressed with mismatched filters optimized by minimizing peak-sidelobe-level (PSL). Both simulation and experiments were carried out to demonstrate the advantage of this technique. The simulation results demonstrated a possible sidelobe reduction (<-90 dB) with a slightly decrease of the signal-to-noise ratio of less than 1 dB compared with the compression using matched filters alone. The experimental results showed about 14 dB SNR improvement as well as -40 dB sidelobe level when the Barker-13 code excitation with 3-cycle sinusoidal wave carrier was used. PMID- 16714041 TI - Interval estimates for epidemic thresholds in two-sex network models. AB - Epidemic thresholds in network models of heterogeneous populations characterized by highly right-skewed contact distributions can be very small. When the population is above the threshold, an epidemic is inevitable and conventional control measures to reduce the transmissibility of a pathogen will fail to eradicate it. We consider a two-sex network model for a sexually transmitted disease which assumes random mixing conditional on the degree distribution. We derive expressions for the basic reproductive number (R(0)) for one and heterogeneous two-population in terms of characteristics of the degree distributions and transmissibility. We calculate interval estimates for the epidemic thresholds for stochastic process models in three human populations based on representative surveys of sexual behavior (Uganda, Sweden, USA). For Uganda and Sweden, the epidemic threshold is greater than zero with high confidence. For the USA, the interval includes zero. We discuss the implications of these findings along with the limitations of epidemic models which assume random mixing. PMID- 16714043 TI - Confirmation of E. coli among other thermotolerant coliform bacteria in paper mill effluents, wood chips screening rejects and paper sludges. AB - Paper sludges are solid wastes material generated from the paper production, which have been characterized for their chemical contents. Some are rich in wood fiber and are a good carbon source, for example the primary and de-inking paper sludges. Others are made rich in nitrogen and phosphorus by pressing the activated sludge, resulting from the biological water treatments, with the primary sludge, yielding the combined paper sludge. Still, in the absence of sanitary effluents very few studies have addressed the characterization of their coliform microflora. Therefore, this study investigated the thermotolerant coliform population of one paper mill effluent and two paper mill sludges and wood chips screening rejects using chromogenic media. For the first series of analyses, the medium used was Colilert broth and positive tubes were selected to isolate bacteria in pure culture on MacConkey agar. In a second series of analyses, double selective media, based on ss-galactosidase and ss-glucuronidase activities, were used to isolate bacteria. First, the presence of thermotolerant coliforms was detected in low numbers in most water effluents, but showed that the entrance of the thermotolerant coliforms was early in the industrial process. Also, large numbers of thermotolerant coliforms, i.e., 7,000,000 MPN/g sludge (dry weight; d.w.), were found in combined sludges. From this first series of isolations, bacteria were purified on MacConkey medium and identified as Citrobacter freundii, Enterobacter sp, E. sakazakii, E. cloacae, Escherichia coli, K. pneumoniae, K. pneumoniae subsp. rhinoscleromatis, K. pneumoniae subsp. ozaenae, K. pneumoniae subsp. pneumoniae, Pantoea sp, Raoultella terrigena, R. planticola. Second, the presence of thermotolerant coliforms was measured at more than 3,700-6,000 MPN/g (d.w) sludge, whereas E. coli was detected from 730 to more than 3,300 MPN/g (d.w.) sludge. The presence of thermotolerant coliform bacteria and E. coli was sometimes detected from wood chips screening rejects in large quantities. Also, indigenous E. coli were able to multiply into the combined sludge, and inoculated E. coli isolates were often able to multiply in wood chips and combined sludge media. In this second series of isolations, API20E and Biolog identified most isolates as E. coli, but others remained unidentified. The sequences of the 16S rDNA confirmed that most isolates were likely E. coli, few Burkholderia spp, but 10% of the isolates remained unidentified. This study points out that the coliform bacteria are introduced by the wood chips in the water effluents, where they can survive throught the primary clarifier and regrow in combined sludges. PMID- 16714044 TI - Spatial and seasonal variations in the water quality of the Amu Darya River (Central Asia). AB - Although the use of the water resources in Uzbekistan is strongly limited by their quality, it has to be noted that there is a lack of information and data within the international scientific literature with regard to the water chemical characteristics of the Amu Darya River, one of the main water resources in Central Asia. To add to such knowledge, this paper examines the spatial and temporal variation of the water quality of the Amu Darya River in order to assess its degree of degradation and the main causal factors. The functional relationships of the pollutants with respect to the flow regime are investigated. Finally an "opportune temporal window" for water withdrawal for filling the reservoirs, in relation to human consumption, will be indicated. The high salinization levels of the waters are mainly due to the presence of sulphates and chlorine. At the up-stream site salinity, although presenting elevated concentrations, does not exceed palatability levels; after the 450 km point the opportune temporal window for water withdrawal with acceptable salinity values is reduced to the period from May to September. Two main driving forces govern the temporal variation of the salinity of the Amu Darya water: a low drainage density of the area which limits the salt loads induced by the natural runoff processes, and snow and glacier melting in the upper catchment area which promotes dilution of the dissolved salts during the high-flow period. During low-flow periods salinity is strongly influenced by the return of waters used for land washing and irrigation. PMID- 16714045 TI - Biomarker responses in European eel (Anguilla anguilla) exposed to persistent organic pollutants. A field study in the Vaccares lagoon (Camargue, France). AB - A screening of relevant biomarkers was carried out in order to evaluate metabolic and cellular damages in European eels exposed to a non-point source contamination by persistent organic pollutants (POP) such as polycyclic aromatic hydrocarbons (PAH) and organochlorine compounds (OC) in a protected area, the Nature Reserve of Camargue (France). Investigations were focused on metabolic responses including detoxification mechanisms (biotransformation, antioxidant process), energy requirements and enzymatic membrane markers either involved in neuronal conduction (acetylcholinesterase, AChE) or in osmoregulation and energy metabolism (ATPases). The hepatic and muscular glycogen rates seemed to be suitable biomarkers as well as three hepatic activities involved in the protection against oxyradicals: catalase, glutathione peroxidase (SeGPx) and superoxide dismutases (SOD). The muscle and gill ATPases as well as the muscle and brain AChE showed more significant relevance in terms of biomarkers than the biotransformation enzymes: ethoxyresorufine-O-deethylase (EROD) and uridine diphospho-glucuronyl transferase (UDPGT). However, most of these enzymatic activities depend on numerous abiotic factors, which must be taken into account in such a biomarker assessment approach. Our study provides some conclusive elements to approve the use in situ of biomarkers developed from laboratory studies. It also raises a question regarding the location of contaminant impregnation in fish organ, in relation with age, development status or mode of contamination, and its influence on biomarker response. If the relevance of membrane indicators is confirmed, this study provides an original statement of the extent of the ecotoxicological threat for the aquatic species in a protected area, due to the occurrence of POP in the cell membranes. PMID- 16714046 TI - Polychlorinated dibenzo-p-dioxin (PCDD) and dibenzofuran (PCDF) isomer patterns from municipal waste combustion: formation mechanism fingerprints. AB - Polychlorinated dibenzo-p-dioxin (PCDD) and dibenzofuran (PCDF) byproducts can be formed in combustion systems by a variety of mechanisms. While total PCDD/F emissions and, to a lesser extent, homologue distributions from incinerators have been found to vary widely depending on combustion conditions, PCDD/F isomer distributions do not. Formation mechanisms can be grouped into two general categories: condensation of precursors, such as chlorinated phenols, and formation from particulate carbon, termed de novo synthesis. In addition to these mechanisms, chlorination and dechlorination reactions may affect isomer patterns. In this work, isomer patterns from field and laboratory municipal waste combustion samples are compared with computed thermodynamic distributions and those from the following experimental investigations: both gas-phase and metal catalyzed condensation of chlorinated phenols, chlorination of dibenzo-p-dioxin and dibenzofuran, and dechlorination of octachlorodibenzo-p-dioxin and octachlorodibenzofuran. PCDD/F isomer patterns produced by different formation mechanisms in controlled experiments are distinct and robust, largely unaffected by combustion conditions. PCDD isomer patterns from municipal waste combustion are most similar to those produced by CuCl(2)-catalyzed phenol condensation from 10 chlorinated phenols. PCDF isomer patterns are most similar to those produced by chlorination and dechlorination. PMID- 16714047 TI - Quantitative structure-activity-activity and quantitative structure-activity investigations of human and rodent toxicity. AB - Acute toxicity in different biological systems, including humans and rodents in vivo, and human and rodent cell lines in vitro, was investigated. The data were taken from the MEIC (Multicentre Evaluation of In Vitro Cytotoxicity) programme. Quantitative structure-activity-activity relationship (QSAAR) models were developed for the in vivo human and rodent toxicity including a combination of toxicity endpoint and structural descriptors as predictor variables. The human peak blood/serum LC(50) concentrations were most strongly related to human liver cell toxicity, while the in vivo oral human lethal doses were most closely related to the in vivo rodent LD(50) values. The QSAARs included structural descriptors encoding electronic/reactivity properties, presence of H-bond donors, compound aromaticity, and size/shape properties. Quantitative structure-activity relationships (QSARs) were derived by using structural descriptors accounting for molecular hydrophobicity, size and shape, and electronic properties. These models have the potential to provide useful insights in the development of non-animal (in vitro and in silico) methods for predicting human and mammalian toxicity. PMID- 16714048 TI - Case report: amyloid tumors in a case of non-secretory multiple myeloma. AB - Multiple myeloma (MM) is a neoplastic disorder characterized by proliferation of a single clone of plasma cells derived from B cells, which proliferates in the bone marrow and frequently invades the adjacent bone, producing skeletal destruction that results in bone pain and fractures. Patients with MM can furthermore present with anemia, hypercalcemia and renal failure. Non-secretory multiple myeloma (NSMM) is characterized by the absence of a monoclonal (M) protein in both the serum and urine. The reported incidence is 1-5% of all multiple myeloma cases. Development of amyloid tumors in NSMM has been described in the literature only occasionally. The clinical features of a 49-year-old female patient with NSMM and amyloid tumors in the breast, lung and rib are presented in this report. Conventional histology, Congo red staining with and without potassium permanganate pretreatment, aldehyde bisulfite-toluidine blue (ABT) reaction, sialic acid specific topo-optical reaction, toluidine blue topo optical reaction as well as immunohistochemistry were performed. An attempt is made to explain the lack of monoclonal immunoglobulins in the serum and urine, although extensive organ amyloidosis of AL type (kappa-light chains) has been found. It is assumed that the plasmocytic plasma cells possess an excretory mechanism, which allows the pathologic immunoglobulins to be secreted either as amyloid proteins polymerizing into amyloid fibrils, or as immunoglobulin fragments that are subject to degradation as soon as they are excreted out of the tumor cell. In this paper, we review the occurrence of amyloid tumors in non secretory multiple myeloma and, in a single case report, we confirm the existence of carbohydrate residues, including sialic acids and sulfated GAGs, in amyloid deposits. PMID- 16714049 TI - Effects of melatonin on streptozotocin-induced diabetic liver injury in rats. AB - This study investigated the possible protective effects of melatonin as an antioxidant against streptozotocin (STZ)-induced diabetic liver injury in rats. Wistar rats were divided into four groups: untreated control (UC), melatonin treated control (MC), untreated diabetic (UD), and melatonin-treated diabetic (MD). Experimental diabetes was induced by a single-dose (60 mg/kg, intraperitoneally (ip)) STZ injection, and melatonin was injected (200 microg/kg/day, ip) for 4 weeks. Upon light and electron microscopic examination, we observed that melatonin improved the morphological and histopathological changes of the liver caused by diabetes. Malondialdehyde levels in the liver homogenates of UD rats were higher than those of controls and were markedly reduced after melatonin treatment. Although no significant difference was observed with respect to antioxidant status, the superoxide dismutase activity tended to be higher in the UD rats than in the treated rats. Our findings showed that melatonin administration partially reduced liver injury in STZ-induced diabetic rats. PMID- 16714050 TI - Monitoring health by values of acute phase proteins. AB - A systemic acute phase reaction may develop during infection and inflammation, due to the action of peripherally liberated proinflammatory cytokines. Hepatic metabolism changes, and negative and positive acute phase proteins (APPs) can be measured in the blood: the APPs therefore represent appropriate analytes to assess health. While they are non-specific markers, their levels change with biological effects and this can be used to assess nutritional deficits and reactive processes, especially when positive and negative acute phase variables are combined in an index. Unfortunately, at present, no comprehensive, easy-to use and cheap system is available to assess various acute phase proteins in serum or blood samples. Protein micro-array technology may satisfy this need; it will permit simultaneous analysis of numerous analytes in the same small volume sample and enable integration of information derived from systemic reactivity and nutrition with disease-specific variables. Applying such technology may help to address health problems in many countries. PMID- 16714051 TI - Sensitivity and specificity of Congo red staining according to Romhanyi. Comparison with Puchtler's or Bennhold's methods. AB - The sensitivity and specificity of various Congo red staining methods is very important in the diagnosis of amyloidosis. When using a less sensitive staining method, some true positive cases of amyloidosis remain undetected. A more highly specific method potentially detects more cases and reveals amyloidosis in an earlier stage of deposition. In this paper, the Congo red staining method according to Romhanyi is discussed in comparison with Puchtler's and Bennhold's methods. Using Romhanyi's technique, there is no alcoholic differentiation, and thus no dye molecules are washed off the amyloid filaments. The binding of the oriented dye molecules is optimal for polarization microscopy. With this method, the polar hydrophilic mounting medium, gum Arabic is used. Mounted in this carbohydrate-containing, hydrophilic medium, the Congo red molecules are oriented parallel to the surface of the amyloid filaments and the sign is linear positive, corresponding to an additive character of topo-optical staining reactions. Otherwise, the Congo red molecules are oriented perpendicular to the surface of collagen, reducing the intensity of birefringence and even inducing an inversion of the original sign of the collagen birefringence. With alcoholic differentiation, Congo red dye molecules are extracted and this decreases the birefringence of amyloid deposits, i.e. minimal amyloid deposits may be missed. Using the apolar hydrophobic mounting medium, Canada balsam, an axis-parallel arrangement of Congo red dye molecules on the surface of collagen fibers and amyloid will occur, resulting in an additive topo-optical reaction with a green polarization color and a false positive diagnosis of amyloidosis ("phantom amyloidosis"). PMID- 16714052 TI - Apomyoglobin reveals a random-nucleation mechanism in amyloid protofibril formation. AB - Protofibrils (PFs) represent the earliest fibrillar species that occur in the course of amyloid fibril formation. Using apomyoglobin, we report here that PFs arise from a multi-step reaction and that they are preceded by an ensemble of non fibrillar particles (NFPs). These intermediate aggregates encompass nascent elements of amyloid structure and can act as seeds in PF formation. Taken together with the observation that PFs often protrude from NFPs, our data suggest that PFs form by a random nucleation mechanism in which the polypeptide chains sample many different aggregated conformations. Once the appropriate structural characteristics are acquired, PFs are formed by addition of further polypeptide chains. PMID- 16714053 TI - Identification of elastic fibers and lamellae in porcine pericardium and aorta by confocal, fluorescence and polarized light microscopy. AB - Pericardial connective tissue has been used to construct bioprostheses to repair various types of injuries, including aortic wall repairs. The arrangement and the distribution of elastic and collagen staple fibers are related directly to the biomechanical properties of the tissue and thus determine the choice tissue for the construction of bioprostheses. Although elastic fibers can be visualized using several histochemical methods, the specificity and mechanism of binding involved remain to be clarified. In this work, we compared the elastic net of the porcine pericardial matrix with that of the aortic wall using 1-anilino-8 naphthalene sulfonate (ANS) and dansyl chloride (DCl) as fluorescent probes and the permanganate-bisulfite-toluidine (PBT) method. Polarized light and fluorescence microscopy were simultaneously used to analyze the tissues. Some samples also were examined using confocal microscopy. Aorta and pericardium treated with ANS and DCl showed elastic fibers and lamellae with an intense blue fluorescence. When stained with the PBT method, the aortic elastic lamellae were clearly metachromatic and, under polarized light, they showed a greenish birefringence. DCl provided clearer fluorescent labelling of elastic fibers when examined using confocal microscopy. PMID- 16714054 TI - Immunoreactive TSH cells in juvenile and peripubertal rats after estradiol and human chorionic gonadotropin treatment. AB - Different hormones and growth factors control the homeostasis of the anterior pituitary gland. We examined the morphological features of pituitary thyroid stimulating hormone (TSH)-producing cells in juvenile and peripubertal female rats after treatments with estradiol-dipropionate (EDP), human chorionic gonadotropin (hCG) and a combination of both hormones (EDP+hCG). TSH-producing cells were labelled using a peroxidase-antiperoxidase immunohistochemical procedure for binding of a rabbit anti-rat beta-thyrotropic polyclonal antisera. Morphometric differences in the cytoplasmic and nuclear volume densities in thyrotropes after the treatments were determined using a stereological method. The relative weights of the pituitary glands were significantly higher in the EDP and EDP+hCG-treated juvenile and peripubertal rats than in untreated age-matched controls. Treatment with EDP promoted a decrease, and treatment with hCG an increase, of the cellular and nuclear volumes in TSH cells in both juvenile and peripubertal females in comparison with the respective controls. Treatment with a combination of EDP+hCG did not induce any significant changes. The cytoplasmic and nuclear volume densities in TSH cells in the EDP+hCG-treated group were significantly higher than in the EDP-treated, and significantly lower than in hCG treated rats at both growth stages. These findings suggest that estradiol and hCG exerted opposite effects on pituitary TSH-immunoreactive cells. The observed effects on thyrotrope morphology were apparently independent of the stage of development. PMID- 16714055 TI - Subcutaneous fat tissue for diagnosis and studies of systemic amyloidosis. AB - The systemic amyloidoses comprise a biochemically heterogeneous group of potentially lethal disorders. An early and precise diagnosis is crucial for the treatment and prognosis. Subcutaneous fat biopsy is a simple and safe method to obtain a diagnosis of systemic amyloidosis and the material can be used for exact determination of amyloid type. A method is described for immunochemical typing of the amyloid based on Western blot analysis combined with specific amyloid fibril protein antibodies. PMID- 16714056 TI - What is the optimal goal of primary cytoreductive surgery for bulky stage IIIC epithelial ovarian carcinoma (EOC)? AB - OBJECTIVE: Recent studies have suggested that the definition of optimal cytoreduction for advanced EOC should be changed from the current Gynecologic Oncology Group threshold of < or =1 cm residual disease to no gross residual disease owing to improved survival of patients (pts) rendered macroscopically disease-free. The objective of this study was to analyze survival rates at very specific residual disease diameters to determine the optimal goal of primary cytoreduction for bulky stage IIIC EOC. METHODS: A prospectively kept database was used to identify and review the records of all pts with Stage IIIC EOC who underwent primary cytoreductive surgery at our institution between January 1989 and December 2003. To analyze a homogeneous cohort of cases, we excluded pts with stage IIIC disease based on nodal metastasis alone (without bulky abdominal tumor), fallopian tube or primary peritoneal carcinomas, and borderline tumors. Standard statistical analyses were utilized. RESULTS: The study cohort included 465 pts. The median age was 60 years (range, 25-87), and the median follow-up was 38 months (range, 1-199). Univariate and multivariate analyses, which included various prognostic factors, identified amount of residual disease as a significant prognostic factor (P < 0.001). Median overall survival in relation to the 5 residual disease categories was: no gross residual, 106 months; gross < or =0.5 cm, 66 months; 0.6-1.0 cm, 48 months; 1-2 cm, 33 months; >2 cm, 34 months. Statistical comparison between the 5 residual disease categories revealed 3 distinct groups with significantly different survival rates (P < 0.01). These 3 groups were: (1) no gross residual; (2) gross < or =1 cm residual; and (3) >1 cm residual. Although the difference in survival did not reach statistical significance, within the gross < or =1 cm residual group, there was a trend toward improved survival in pts left with smaller volume, < or =0.5 cm residual compared with those with 0.6-1.0 cm residual (P = 0.06). CONCLUSION: Our data suggest that removal of all evidence of macroscopic disease is associated with prolonged survival and should be the goal of primary cytoreductive surgery. If complete gross resection is not feasible, however, cytoreduction to as minimal residual tumor as possible should be the focus of cytoreductive efforts, as each incremental decrease in residual disease below 1 cm may be associated with an incremental improvement in overall survival. PMID- 16714057 TI - Serum gamma glutamyltransferase as a marker of metabolic syndrome and coronary disease likelihood in nondiabetic middle-aged and elderly adults. AB - The role of serum gamma glutamyltransferase (GGT) activity as a cardiovascular risk marker was studied basically cross-sectionally. After appropriate exclusions, 754 men and 802 women were available for analysis who were followed up briefly yielding only 16% of overall cases of coronary heart disease (CHD). GGT activity was measured kinetically. In multivariate analysis across 12 variables, waist circumference, sex, complement C3, moderate alcohol intake and uric acid were significant independent covariates of serum GGT. By analyzing the sample in tertiles, doubling in GGT activity was found associated with a rise of 74% in metabolic syndrome (MS) likelihood-independent of salient confounders (P < 0.001). This association was mediated by waist circumference. Individuals in the top versus the bottom tertile exhibited an odds ratio for CHD likelihood of 1.81 (95% CI 1.09; 3.02)-independent of age, sex, total cholesterol, systolic blood pressure, impaired fasting glucose, smoking status, alcohol usage and, notably, of waist circumference. This indicated that a doubling in serum GGT activity corresponded to a 45% excess in CHD likelihood, after adjustment for standard risk factors. In conclusion, waist circumference is a major determinant of serum GGT activity among Turkish adults. Doubling in activity is associated with a (largely waist girth mediated) rise by over one-half in the multiadjusted MS likelihood, and by nearly one-half in the CHD likelihood, independent of waist girth and major risk factors. PMID- 16714058 TI - Alginate-whey protein granular microspheres as oral delivery vehicles for bioactive compounds. AB - Alginate (AL)-whey protein isolate (WPI) microspheres of varied WPI/AL ratio, particle diameter and concentration of polymer bead forming solution (C(AL+WPI)) were prepared in order to develop a biocompatible vehicle for oral administration of bioactive compounds. Microscopy revealed a special matrix/granular structure for microspheres with a WPI/AL ratio of 8:2, 100 microm diameter and C(AL+WPI) of 5% (AL-WPI A2), featuring WPI granules 3-10 microm in diameter homogeneously distributed within an AL spherical matrix. The compound release properties of these microspheres were investigated in simulated gastric and intestinal fluids (SGF and SIF). They demonstrated the desirable property of retarding riboflavin release in SGF and underwent alginate matrix erosion together with liberation of WPI granules in SIF, followed by complete release of the riboflavin. Riboflavin release in SGF and in SIF without pancreatin followed the Higuchi diffusion model while release in SIF in the presence of pancreatin was attributed to WPI granule degradation. PMID- 16714059 TI - Expression of the fusogenic HERV-FRD Env glycoprotein (syncytin 2) in human placenta is restricted to villous cytotrophoblastic cells. AB - Recently, the expression of a human endogenous retrovirus HERV-FRD, able to encode a fusogenic envelope protein (syncytin 2), has been observed in human placenta. The aim of the present study was to localize the expression of syncytin 2 in first trimester placenta. In addition, we investigated the presence of HERV FRD transcripts during the in vitro differentiation of isolated villous and extravillous trophoblastic cells from first trimester chorionic villi. Using a monoclonal antibody specifically raised against the HERV-FRD Env protein, syncytin 2 was immunolocalized only in the villous trophoblast of the chorionic villi, at the level of cytotrophoblastic cells. Interestingly, immunostaining was not observed in all cells but only in some of them, and was detected, more frequently, at the membrane level at the interface between the cytotrophoblastic cells and syncytiotrophoblast. Labeling was observed neither in the syncytiotrophoblast nor in the mesenchymal core of the villi nor in the extravillous trophoblast. In vitro detection of HERV-FRD transcripts was restricted to villous trophoblastic cells and decreased significantly with time in culture. These results suggest that syncytin 2 might play a role in human trophoblastic cell fusion. PMID- 16714060 TI - V(H)3-48 and V(H)3-53, as well as V(H)3-21, gene rearrangements define unique subgroups in CLL and are associated with biased lambda light chain restriction, homologous LCDR3 sequences and poor prognosis. AB - This study determined IgV(H) gene usage in 228 chronic lymphocytic leukaemia patients to investigate associations between gene usage and other biological or clinical characteristics. V(H)3-48 [N=8] and V(H)3-53 [N=4] gene rearrangements showed biased lambda light chain restriction and were predominantly found in female patients with short lymphocyte doubling time but without adverse prognosis cytogenetics. Overuse of V(L)3-21(Vlambda2-14) gene and highly homologous LCDR3 sequences were found in V(H)3-48 patients. V(H)3-21 gene usage [N=18, 7.9%] was associated with poor prognosis, overuse of V(L)3-21(Vlambda2-14) gene and highly homologous heavy- and light-chain CDR3 sequences, but was not associated with poor prognosis chromosomal aberrations. PMID- 16714061 TI - Assessing the proarrhythmic potential of drugs: current status of models and surrogate parameters of torsades de pointes arrhythmias. AB - Torsades de pointes (TdP) is a potentially lethal cardiac arrhythmia that can occur as an unwanted adverse effect of various pharmacological therapies. Before a drug is approved for marketing, its effects on cardiac repolarisation are examined clinically and experimentally. This paper expresses the opinion that effects on repolarisation duration cannot directly be translated to risk of proarrhythmia. Current safety assessments of drugs only involve repolarisation assays, however the proarrhythmic profile can only be determined in the predisposed model. The availability of these proarrhythmic animal models is emphasised in the present paper. It is feasible for the pharmaceutical industry to establish one or more of these proarrhythmic animal models and large benefits are potentially available if pharmaceutical industries and patient-care authorities embraced these models. Furthermore, suggested surrogate parameters possessing predictive power of TdP arrhythmia are reviewed. As these parameters are not developed to finalisation, any meaningful study of the proarrhythmic potential of a new drug will include evaluation in an integrated model of TdP arrhythmia. PMID- 16714062 TI - Pharmacokinetic and pharmacodynamic alterations of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors: drug-drug interactions and interindividual differences in transporter and metabolic enzyme functions. AB - 3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) are widely used for the treatment of hypercholesterolemia. Their efficacy in preventing cardiovascular events has been shown by a large number of clinical trials. However, myotoxic side effects, sometimes severe, including myopathy or rhabdomyolysis, are associated with the use of statins. In some cases, such toxicity is associated with pharmacokinetic alterations. In this review, the pharmacokinetic aspects and physicochemical properties of statins are reviewed in order to understand the mechanism governing their pharmacokinetic alterations. Among the statins, simvastatin, lovastatin and atorvastatin are metabolized by cytochrome P450 3A4 (CYP3A4) while fluvastatin is metabolized by CYP2C9. Cerivastatin is subjected to 2 metabolic pathways mediated by CYP2C8 and 3A4. Pravastatin, rosuvastatin and pitavastatin undergo little metabolism. Their plasma clearances are governed by the transporters involved in the hepatic uptake and biliary excretion. Also for other statins, which are orally administered as open acid forms (i.e. fluvastatin, cerivastatin and atorvastatin), hepatic uptake transporter(s) play important roles in their clearances. Based on such information, pharmacokinetic alterations of statins can be predicted following coadministration of other drugs or in patients with lowered activities in drug metabolism and/or transport. We also present a quantitative analysis of the effect of some factors on the pharmacokinetics of statins based on a physiologically based pharmacokinetic model. To avoid a pharmacokinetic alteration, we need to have information about the metabolizing enzyme(s) and transporter(s) involved in the pharmacokinetics of statins and, along with such information, model-based prediction is also useful. PMID- 16714064 TI - The problem of neuronal cell types: a physiological genomics approach. AB - Neural circuits within the vertebrate brain are composed of highly diverse cell types. The exact extent of this diversity is a matter of continuing debate. For example, do cortical interneurons comprise a few, dozens or >100 distinct cell types? Recently, several groups have used microarrays to measure genome-wide gene expression profiles for specific neuronal cell types. These methods can offer an objective basis for neuronal classification. In this review, we argue that this approach should now be carried out more broadly and that it should be coupled to large-scale efforts to generate mouse driver lines in which tools for genetic manipulation, such as the Cre recombinase, are expressed in identified cell types within the brain. This would enable neuroscientists to begin to investigate more systematically the roles of specific genes in establishing particular cellular phenotypes, and also the roles of particular cell types within brain circuits. This review is part of the TINS special issue on The Neural Substrates of Cognition. PMID- 16714063 TI - Porcine gammadelta T cells: possible roles on the innate and adaptive immune responses following virus infection. AB - gammadelta T cells recognise different types of antigen in alternative ways to alphabeta T cells, and thus appear to play a complementary role in the immune response. However, unlike alphabeta T cells, the role or function of gammadelta T cells is still unclear. As pigs possess a high proportion of circulating gammadelta T cells, they are suitable large animal model to study gammadelta T cell functions. This as yet has not been fully exploited, leaving porcine gammadelta T cell biology and its role in immunity in its infancy. Foot-and-mouth disease (FMD) high potency "emergency" vaccines are able to induce early protection from challenge and it has been suggested that, in part, there is some involvement of innate immune responses. The antigen component of the vaccine is able to stimulate purified naive pig gammadelta T cells and induce the mRNA of various cytokines and chemokines. This observation suggests that gammadelta T cells probably contribute to the early phase of the immune responses to FMD vaccination, and perhaps infection. A subset of these circulating gammadelta T cells display a phenotype similar to professional antigen presenting cells and are able to take up and present soluble antigen to CD4(+) T cells in a direct cell-cell interaction via MHC class II. This direct interaction between gammadelta T cells and CD4(+) T cells is likely to have a significant influence on the out come of the adaptive immune response. PMID- 16714065 TI - Stability of casein mRNA is ensured by structural interactions between the 3' untranslated region and poly(A) tail via the HuR and poly(A)-binding protein complex. AB - The maintenance of mRNA stability has emerged as a mechanism of post transcriptional control. We demonstrate that beta-casein mRNA stability is dictated by the poly(A) tail and sequences in the 3'-UTR. An in vitro mRNA decay assay revealed that beta-casein mRNA with a long poly(A) tail had higher stability than that with a short poly(A) tail. The addition of poly(A) homopolymer and 3'-UTR cRNA as competitor induced rapid degradation of beta casein mRNA. The interaction between full-length beta-casein mRNA and poly(A) homopolymer was inhibited by the addition of the 3'-UTR cRNA in the lysates, which indicates that one region of the 3'-UTR associates with the poly(A) tail through an RNA-protein interaction. The putative AU-rich element (ARE) is present at nt 897-905; deletion and mutation analysis showed that the ARE site was required for maintaining the stability of the beta-casein 3'-UTR. In the immunoprecipitation analysis, the poly(A)-binding protein (PABP) and the RNA binding protein HuR were pulled down by 3'-UTR cRNA, and the absence of the ARE site reduced the binding of these proteins. These experiments further revealed that the rapid degradation of beta-casein mRNA was induced by incubation with HuR and PABP-depleted RRLs. Collectively, our results suggest that beta-casein mRNA is protected from degradation by virtue of the structural interaction between the 3'-UTR and poly(A) tail via a protein complex of HuR and PABP. PMID- 16714066 TI - Transient increase of QRS amplitude during coronary artery angiography--a case report. PMID- 16714067 TI - Relationships between dual-task related changes in stride velocity and stride time variability in healthy older adults. AB - Dual-task related gait changes have been previously reported for healthy older adults, suggesting that gait control requires attention. Compared to balance control, the involvement of attention in the control of the rhythmic stepping mechanism, as reflected by stride time variability, is not well known. In particular, under dual-task, the relative contributions of a second, attention demanding task and changes in walking speed remain unclear. Thus, the aims of this study were (1) to assess whether walking with a slow-selected speed or walking while performing an attention-demanding task affected stride time variability in a sample of healthy older participants, and (2) to establish whether stride time variability under dual-task conditions is related either to the decrease of walking speed or the simultaneous attention-demanding task, or to both. Forty-five healthy older participants performed four experimental conditions: (1) walking at a normal self-selected speed, (2) walking at a slow self-selected speed, (3) performing a verbal fluency task when sitting on a chair, and (4) performing the verbal fluency task while walking at self-selected walking speed. Gait parameters were recorded across 15 meters, using Physilog. Results showed a significant dual-task related decrease in mean values of stride velocity, as well as a significant increase in mean values and coefficients of variation of stride time. These dual-task related changes in stride time were explained by the simultaneous performance of the verbal fluency task, the decrease of gait speed and the variability between participants. Although a relationship exists between decreased walking speed and increased stride time variability, the dual-task related increase of stride time variability was also significantly associated with the attention-demanding task, suggesting some attentional control for the rhythmic stepping mechanism of walking in healthy older adults. PMID- 16714068 TI - The role of processing on ochratoxin A content in Italian must and wine: a study on naturally contaminated grapes. AB - Available data show that ochratoxin A (OTA) is a possible contaminant of wine and its concentration is higher in red wines than in white and rose. The aim of this work was to study the fate of OTA during the main stages of the winemaking process (crushing, maceration, alcoholic fermentation, malo-lactic fermentation, bottle-aging) and the influence of technological treatments on OTA concentration in order to identify the critical process steps. Attention was focused on red winemaking, in different wineries in the south of Italy, with two naturally OTA contaminated grape varieties ('Negroamaro' and 'Primitivo') and on different vintages (2001-2002-2003). The results show that no OTA is produced during winemaking, but each operation during winemaking can modify OTA content. The OTA present in grapes to a certain degree is released to the juice during crushing. Maceration increases the OTA content, while alcoholic and malo-lactic fermentation cause a reduction in OTA in the wine. PMID- 16714069 TI - Eicosapentaenoic acid confers neuroprotection in the amyloid-beta challenged aged hippocampus. AB - Among the changes that occur in the hippocampus with age, is a deficit in long term potentiation (LTP). This impairment is associated with inflammatory changes, which are typified by increased concentration of the pro-inflammatory cytokine interleukin-1beta (IL-1beta). Activated microglia are the most likely cell source of IL-1beta, but data demonstrating an age-related increase in microglial activation is equivocal. Here we demonstrate that the age-related deficit in LTP is accompanied by increased expression of cell surface markers of activated microglia (major histocompatibility complex II and CD40) and increased IL-1beta production, and that these changes may be stimulated by interferon-gamma. Treatment of aged rats with eicosapentaenoic acid (EPA) attenuates these changes and we suggest that IL-4 mediates the action of EPA. We demonstrate that aged rats exhibit an exaggerated response to intracerebroventricular injection of beta amyloid peptide 1-40 (Abeta). Thus Abeta inhibited LTP in aged, but not young, rats and induced a further increase in hippocampal IL-1beta concentration. Of particular significance is the demonstration that EPA protects the aged brain so that the increased vulnerability to Abeta is ameliorated in EPA-treated rats. PMID- 16714071 TI - The cell-mediated immune response induced by plasmid encoding bovine herpesvirus 1 glycoprotein B is enhanced by plasmid encoding IL-12 when delivered intramuscularly or by gene gun, but not after intradermal injection. AB - Bovine herpesvirus 1 (BHV-1) causes respiratory and genital infections in cattle. Previously we demonstrated that a DNA vaccine encoding a truncated, secreted form of BHV-1 glycoprotein B (tgB) induces cytotoxic T lymphocyte (CTL) responses in C3H mice. In this study we investigated the potential of interleukin 12 (IL-12) to further enhance the CTL response. C3H mice were immunized with a plasmid encoding tgB or with plasmids encoding tgB and murine IL-12. When the plasmid encoding tgB was delivered intramuscularly or epidermally by a gene gun, co administration with IL-12 plasmid stimulated the synthesis of more IgG2a, the production of higher levels of IFN-gamma, and more effective killing by CTLs. In contrast, after intradermal delivery no effect of co-administration of IL-12 encoding plasmid was observed. Further investigation suggested that antigen and IL-12 need to be expressed in the draining lymph nodes, where IL-12 can have a direct effect on T cells. PMID- 16714070 TI - Mucosal immunization against hepatitis A: antibody responses are enhanced by co administration of synthetic oligodeoxynucleotides and a novel cationic lipid. AB - Hepatitis A caused by hepatitis A virus (HAV) transmitted by the fecal-oral route, results in considerable morbidity and economic loss. Mucosal immunization can be more effective than conventional injection at inducing both local and systemic immunity to HAV. Here we show that co-administration of killed HAV with synthetic oligodeoxynucleotides (ODNs) containing CpG sequences, and a novel polycationic sphingolipid (CCS)/cholesterol liposomal delivery system, markedly enhances the HAV-specific antibody response at the intestinal interface, particularly when delivered intrarectally or intranasally, to Balb/c mice at low HAV doses. A mucosally delivered, antigen-sparing HAV vaccine that is easily administered without specialized equipment or personnel, is an attractive alternative for facilitating mass immunization in hepatitis A outbreaks. PMID- 16714072 TI - Generation of anti-tumor immunity using mammalian heat shock protein 70 DNA vaccines for cancer immunotherapy. AB - In this study, we explored the protective anti-tumor potency of mouse (self) Hsp70 or Hsp110-based DNA vaccination approach targeting a tumor-associated antigen, human papilloma virus (HPV) type 16 E7 protein. Linkage of E7 to the N terminus of the mouse Hsp70 not only elicits an E7-specific cytotoxic T cell (CTL) response, but also protects mice against challenge with E7 expressing tumors. CD8+ T-cells are crucial in both priming and effector phases for the induction of tumor immunity, whereas CD4+ T-cells and NK cells do not appear to play a major role. Furthermore, the ATP-binding domain deletion mutant Hsp70(382 641), when fused to E7, was immunologically effective, suggesting that the peptide-binding region, not the ATPase domain of Hsp70, is required for the vaccine activity of the E7-Hsp70 DNA. This study demonstrates that autologous Hsp70 is highly potent in enhancing antigen-specific immune responses. Functional domain mapping and orientation of the E7 and Hsp70 in the fusion gene may have clinical implications for the design and optimization of Hsp70-based DNA vaccines. PMID- 16714074 TI - Re: the safest location for steroid injection in the treatment of carpal tunnel syndrome. PMID- 16714073 TI - Importance of HLA-DQ and HLA-DP polymorphisms in cytokine responses to naturally processed HLA-DR-derived measles virus peptides. AB - We studied the association between class II human leukocyte antigen (HLA) DRB1*0301 presented measles virus (MV) peptide-specific cytokine responses and DQB1 and DPB1 alleles among 313 individuals who received two doses of measles mumps-rubella-II vaccine. The overall median IFN-gamma secretion levels (first and third quartiles) for the 19-amino acid MV phosphoprotein (MV-P)- and 14-amino acid MV nucleoprotein (MV-N)-derived peptides were 27.7 pg/ml (1.8, 109.4) and 1.9 pg/ml (-6.2, 13.0), respectively; median IL-4 secretion levels were -0.6 pg/ml (-7.1, 6.2) and 2.4 pg/ml (-3.2, 9.3), respectively. Primary statistical analyses were adjusted for previously identified DRB1 associations. A marginally significant increase in the frequency of the DQB1*0604 (p=0.02) allele was found among subjects who demonstrated detectable IL-4 levels to the MV-P peptide. Further, DPB1*0201 (p=0.02) and DPB1*1301 (p=0.09) alleles provided suggestive evidence of an association with MV-P-induced IL-4 secretion. Examination of IFN gamma responses to MV-P and MV-N indicated that none of the individual alleles of the DQB1 and DPB1 loci were associated with peptide-induced T cell response. An increase in the frequency of DPB1*0501 (p=0.01) was found among subjects who failed to produce MV-N peptide-specific IL-4 responses. These data further confirm that HLA-DRB1 alleles are the major restriction molecules for MV-P and MV N measles virus antigen presentation to T cells. We speculate that MV-P and MV-N peptides derived from DRB1*0301 could potentially be recognized in association with different HLA molecules, including DQB1 and DPB1; however, statistical adjustments for the effect of HLA-DR locus could potentially alter these genetic relationships. This concept provides important information supporting the use of promiscuous peptides in a peptide-based vaccine approach. PMID- 16714075 TI - A comparison of blue crab and bivalve delta15N tissue enrichment in two North Carolina estuaries. AB - Stable isotope analyses (delta(15)N) were used to examine invertebrate tissue enrichment in two North Carolina estuaries with differing amounts of nutrient loading. Bivalves collected from a nutrient sensitive estuary yielded a significant difference in mean nitrogen isotopic composition of tissue (10.4 per thousand+/-0.82; N=66) compared to bivalves collected from a less nutrient sensitive estuary (6.4 per thousand+/-0.63; N=45). Similarly, blue crabs from nutrient sensitive sites had a nitrogen isotopic composition of 11.4 per thousand (+/-1.3, N=77), which was significantly different (P<0.001) than the tissue of less nutrient sensitive blue crabs (9.6 per thousand+/-0.6; N=77). The results showed that an inverse relationship exists between invertebrate tissue enrichment and indicators of water quality across estuarine sites. This study suggests that a relationship may exist between nutrient sources and subsequent energy transfer to estuarine consumers in two North Carolina estuaries. PMID- 16714076 TI - Distribution of elements in needles of Pinus massoniana (Lamb.) was uneven and affected by needle age. AB - Macronutrients (P, S, K, Na, Mg, Ca), heavy metals (Fe, Zn, Mn, Cu, Pb, Cr, Ni, Cd) and Al concentrations as well as values of Ca/Al in the tip, middle, base sections and sheaths of current year and previous year needles of Pinus massoniana from Xiqiao Mountain were analyzed and the distribution patterns of those elements were compared. The results indicated that many elements were unevenly distributed among the different components of needles. Possible deficiency of P, K, Ca, Mn and Al toxicity occurred in needles under air pollution. Heavy metals may threaten the health of Masson pine. Needle sheaths were good places to look for particulate pollutants, in this case including Fe, Cu, Zn, Pb, Cr, Cd and Al. PMID- 16714077 TI - In vitro studies of copper release from powder particles in synthetic biological media. AB - The aim of this paper is to provide quantitative data on copper release from powder particles of different copper materials, including artificial copper patina, Cu(2)O and metallic Cu, when exposed to different synthetic biological media to simulate an inhalation scenario and/or skin contact. Generated data may contribute in risk assessment of potential health effects following exposure to and handling of various copper materials. All tests were performed in vitro to determine total copper concentrations, release rates of total copper, and to elucidate its time-dependence. The copper release process was interpreted in terms of specific surface area, surface morphology-, and composition. All powder materials show a time-dependent release process with total copper release rates less than 3 microg/cm(2) per hour at steady state conditions, for all media investigated. The importance of using relevant test media when simulating different interstitial lung conditions and difficulties encountered when comparing powder particles of essentially different properties are thoroughly discussed. PMID- 16714078 TI - Response of epiphytic lichen communities to decreasing ammonia air concentrations in a moderately polluted area of The Netherlands. AB - Decreasing local ammonia air concentrations in a moderately polluted area in The Netherlands were accompanied by a rapid increase in nitrogen-sensitive species (acidophytes) and a decline of nitrogen-tolerant macrolichens (nitrophytes). This paper presents data on the relationship between nitrophyte abundance and species abundance for three ecological groups of epiphytic lichens: nitrophytes (positively correlated with ammonia), acidophytes (negatively correlated) and neutrophytes (which have an optimum at medium concentrations) and suggests ammonia dependent optimum curves for these groups. In this study neutrophytes were found to die-off massively at sites with a decrease of the ammonia air concentration over the period 1996-2003. PMID- 16714079 TI - An integrated impact assessment and weighting methodology: evaluation of the environmental consequences of computer display technology substitution. AB - Computer display technology is currently in a state of transition, as the traditional technology of cathode ray tubes is being replaced by liquid crystal display flat-panel technology. Technology substitution and process innovation require the evaluation of the trade-offs among environmental impact, cost, and engineering performance attributes. General impact assessment methodologies, decision analysis and management tools, and optimization methods commonly used in engineering cannot efficiently address the issues needed for such evaluation. The conventional Life Cycle Assessment (LCA) process often generates results that can be subject to multiple interpretations, although the advantages of the LCA concept and framework obtain wide recognition. In the present work, the LCA concept is integrated with Quality Function Deployment (QFD), a popular industrial quality management tool, which is used as the framework for the development of our integrated model. The problem of weighting is addressed by using pairwise comparison of stakeholder preferences. Thus, this paper presents a new integrated analytical approach, Integrated Industrial Ecology Function Deployment (I2-EFD), to assess the environmental behavior of alternative technologies in correlation with their performance and economic characteristics. Computer display technology is used as the case study to further develop our methodology through the modification and integration of various quality management tools (e.g., process mapping, prioritization matrix) and statistical methods (e.g., multi-attribute analysis, cluster analysis). Life cycle thinking provides the foundation for our methodology, as we utilize a published LCA report, which stopped at the characterization step, as our starting point. Further, we evaluate the validity and feasibility of our methodology by considering uncertainty and conducting sensitivity analysis. PMID- 16714080 TI - Iglesias resectoscope with simultaneous irrigation, suction and low intravesical pressure. 1975. PMID- 16714081 TI - Long-lasting spontaneous calcium transients in the striatal cells. AB - The striatum plays an important role in linking cortical activity to basal ganglia output. We conducted the calcium (Ca2+) imaging to investigate the spontaneous activities of the striatum using acute slice preparations. Corticostriatal slices of rat brain were stained with Fura-PE3-AM. Long-lasting spontaneous intracellular Ca2+ ([Ca2+]i) transients, which lasted up to about 250 s, were observed. The amplitudes of the transients were variable even in a single cell. Most cells exhibited irregular frequencies, but some exhibited oscillatory features. These [Ca2+]i transients were not induced by action potentials because they were not inhibited by tetrodotoxin. Antagonists of the ionotropic glutamate receptors, 6-cyano-7-nitroquinoxaline-2,3-dione and D,L-2-amino-5 phosphonovaleric acid, did not block these transients. These results suggested that the action potentials and the excitatory synaptic inputs in these striatal network were not involved in the induction of the [Ca2+]i transients. In contrast, the number of the active cells, which exhibited the [Ca2+]i transients, was greatly reduced by the intracellular Ca2+ store depletor, thapsigargin. Therefore, the intracellular Ca2+ store is likely to contribute to the [Ca2+]i transients. PMID- 16714082 TI - Immunocytochemical localization of the GABAB2 receptor subunit in the glomeruli of the mouse main olfactory bulb. AB - The olfactory input to the brain is carried out by olfactory nerve axons that terminate in the olfactory bulb glomeruli and make synapses onto dendrites of glutamatergic projection neurons, mitral and tufted cells, and GABAergic interneurons, periglomerular cells. The dendrites are reciprocally connected through asymmetric synapses of mitral/tufted cells with periglomerular cells and symmetric synapses of the opposite direction. Transmission at the first synapse in the olfactory pathway is regulated presynaptically, and this regulation is mediated, in part, by metabotropic GABAB receptors that, when activated, inhibit transmitter release from the olfactory nerve. Functional GABAB receptors are heterodimers composed of the GABAB1 and GABAB2 subunits. Studies using double immunofluorescence have shown colocalization of both subunits in the glomerular neuropil, and ultrastructural studies have localized GABAB1 to extrasynaptic, synaptic, and perisynaptic sites on the plasma membrane of olfactory nerve terminals. We studied the subcellular localization of GABAB2 in the mouse olfactory glomeruli using a subunit-specific antibody and preembedding immunogold labeling. Immunoreactivity for GABAB2 was associated with symmetric dendrodendritic synapses of periglomerular cells with mitral/tufted cells and was localized to the extrasynaptic plasma membrane of presynaptic dendrites, and extrasynaptic, synaptic, and perisynaptic sites on the plasma membrane of postsynaptic dendrites. The results suggest that postsynaptic, and perhaps presynaptic, GABAB receptors may be expressed at GABAergic synapses between dendrites of periglomerular interneurons and projection neurons. Immunolabeling was observed at junctions of the olfactory nerve with mitral/tufted cell dendrites, providing ultrastructural evidence for the expression of the GABAB2 subunit at the primary olfactory synapse. PMID- 16714083 TI - DPCPX-resistant hypoxic synaptic depression in the CA1 region of hippocampal slices: possible role of intracellular accumulation of monocarboxylates. AB - Adenosine plays the principal role in synaptic depression during various energy depleted conditions. However, additional inhibitory factors not associated with A1 adenosine receptors appear to be involved in hypoxic insults. Monocarboxylate accumulation and consequent acidic changes during hypoxia may be responsible for this remaining depression in synaptic activity. Field evoked potentials were recorded in the CA1 region of rat hippocampal slices. Preincubation with 8 cyclopentyl-1,3-dipropylxanthine (DPCPX) disclosed 43% of DPCPX-resistant synaptic depression (DRSD) during oxygen deprivation (OD). In contrast, no DRSD was detected in various conditions with limited glucose utilization, such as glucose deprivation and oxygen-glucose deprivation. Inhibition of anaerobic glycolysis (iodoacetate, sodium fluoride) abolished DRSD during OD, whereas blockade of monocarboxylate utilization with alpha-cyano-4-hydroxycinnamic acid (4-CIN) provoked DRSD in normoxic medium. These observations suggest that an intracellular accumulation of monocarboxylates is responsible for DRSD during hypoxia. PMID- 16714084 TI - Twenty-four hours retention of visuospatial memory correlates with the number of parietal sleep spindles. AB - Recent evidence suggests that the sleep-dependent consolidation of declarative memories relies on the non-rapid eye movement (NREM) rather than the rapid eye movement (REM) phase of sleep. Moreover, a few studies both at the cellular and the behavioural levels have suggested the involvement of sleep spindles, the most synchronous oscillatory waveforms during NREM sleep stage 2, in this process. Our previous study showed that overnight verbal memory retention correlates with the total number of sleep spindles in left frontocentral areas, while spindling in other regions did not correlate with mnemonic retention. In the present study, we show that retention of visuospatial memories over a 24-h period correlates with the total number of sleep spindles detected over parietal regions during the intervening night-time sleep. This result provides further evidence for the association between sleep spindle activity and declarative memory consolidation, and suggests that visuospatial and verbal memory retention differ in the topographic distribution of the NREM spindle activity with which they are associated. PMID- 16714085 TI - Osteoarthritis and joint pain. PMID- 16714086 TI - Attenuation of capsaicin-evoked mechanical allodynia by peripheral neuropeptide Y Y1 receptors. AB - Neuropeptide Y (NPY) and its cognate receptors are important modulators of nociception and their expression is significantly altered following injury. In particular, previous studies have demonstrated that the Y1 subtype of NPY receptors inhibits nociceptive transmission from capsaicin-sensitive terminals in the dorsal horn of the spinal cord. The present study evaluated the function of the Y1 receptor on peripheral terminals of primary afferent neurons by testing whether peripherally administered Y1 agonists and antagonists alter capsaicin evoked mechanical allodynia in rats and capsaicin-evoked immunoreactive calcitonin gene-related peptide (iCGRP) release from isolated superfused rat skin. Treatment with the Y1 agonist [Leu31,Pro34]-NPY (0.5, 1, or 10 nmol) significantly inhibited capsaicin-evoked mechanical allodynia in a dose-dependent manner. This effect was reversible by pretreatment with the Y1 antagonist BIBO3304 (10 nmol). The anti-allodynia produced by the Y1 agonist occurred at a peripheral site of action, because injection into the paw contralateral to the site of the capsaicin injection had no effect on paw withdrawal latencies. In isolated skin, application of [Leu31,Pro34]-NPY (300 nM) significantly inhibited capsaicin-evoked CGRP release. BIBO3304 reversed this inhibition, having itself no effect on capsaicin-evoked iCGRP release. These studies indicate that the activation of peripheral Y1 receptors produces anti-allodynia, possibly via the direct inhibition of capsaicin-sensitive fibers. PMID- 16714087 TI - FBXO25, an F-box protein homologue of atrogin-1, is not induced in atrophying muscle. AB - Atrogin-1/MAFbx/FBXO32 is a muscle-specific ubiquitin-ligase (E3) that is dramatically increased in atrophying muscle. Here, we have investigated the functional relationship between atrogin-1 and FBXO25 which shares 65% amino acid identity. Using a RT-PCR, we demonstrated that FBXO25 is highly expressed in brain, kidney, and intestine, whereas atrogin-1 expression is largely restricted to striate muscle. FBXO25 was shown here to contain a functional F-box domain that binds to Skp1 and thereby to Roc1 and Cul1, the major components of SCF-type E3s. In addition, the productive SCF complex containing FBXO25 showed ubiquitin ligase activity. We investigated the differential expression of atrogin-1 and FBXO25 in fasted and dexamethasone-treated mice and also in rats with streptozotocin-induced diabetes. Although the atrogin-1 was strongly induced in muscle in all three models, no changes were observed in the expression of FBXO25. Therefore, here we have shown that FBXO25 is a novel F-box protein analogous to atrogin-1, which is not involved in muscle atrophy. Further functional studies should elucidate the exact role of FBXO25 in the ubiquitin-proteasome pathway. PMID- 16714088 TI - Esterification of ferulic acid with polyols using a ferulic acid esterase from Aspergillus niger. AB - Commercially available enzyme preparations were screened for enzymes that have a high ability to catalyze direct ester-synthesis of ferulic acid with glycerol. Only a preparation, Pectinase PL "Amano" produced by Aspergillus niger, feruloylated glycerol under the experimental conditions. The enzyme responsible for the esterification was purified and characterized. This enzyme, called FAE PL, was found to be quite similar to an A. niger ferulic acid esterase (FAE-III) in terms of molecular mass, pH and temperature optima, substrate specificity on synthetic substrates, and the N-terminal amino acid sequence. FAE-PL highly catalyzed direct esterification of ferulic acid and sinapinic acid with glycerol. FAE-PL could feruloylate monomeric sugars including arabinose, fructose, galactose, glucose, and xylose. We determined the suitable conditions for direct esterification of ferulic acid with glycerol to be as follows: 1% ferulic acid in the presence of 85% glycerol and 5% dimethyl sulfoxide at pH 4.0 and 50 degrees C. Under these conditions, 81% of ferulic acid could be converted to 1-glyceryl ferulate, which was identified by (1)H-NMR. The ability of 1-glyceryl ferulate to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals was higher than that of the anti-oxidant butyl hydroxytoluene. PMID- 16714089 TI - Clinical experience with Alloderm: a one-staged composite dermal/epidermal replacement utilizing processed cadaver dermis and thin autografts. AB - Alloderm has been advocated for the management of acute burns. However, few studies have demonstrated the feasibility of this technique. METHODS: We reviewed the medical records of all patients treated in our burn center who received Alloderm since 1999. RESULTS: Alloderm was used in 21 burn patients and 6 patients with traumatic skin loss. The average size of Alloderm used in the burn patients was 517+/-144 cm(2) (range 24-3000 cm(2)). The average Alloderm thickness used was 0.008 in. and autografts were harvested at an average of 0.007 in. Overall, Alloderm was used in a variety of locations including the face in 3 patients (2 burns, 1 traumatic skin loss) and hands in 7 patients (6 burns, 1 traumatic skin loss). Successful take was observed in 26/27 patients. CONCLUSIONS: Alloderm can be used successfully in patients with acute burns requiring grafting. PMID- 16714090 TI - Is repetitive wounding and bone marrow-derived stem cell mediated-repair an etiology of lung cancer development and dissemination? AB - The prevailing view of lung cancer is multi-step progression of normal cells into cancer cells through gain of function oncogenes coupled with loss of tumor suppressor genes. This progression of genetic damage ultimately results in the hallmarks of cancer. This theory has strong support from studies finding genetic damage in early stage preneoplastic lesions in lung epithelial cells from current or former smokers. This paper discusses an alternate theory that lung cancer is a bone marrow stem cell derived disease. Chronic cigarette smoking results in lung inflammation and epithelial damage that activates a chronic wound repair program. Recent studies have demonstrated that ability of bone marrow derived stem cells to respond to epithelial wounding and contribute to epithelial repair. The identification of cancer stem cells that are distinct from the bulk tumor cells through their ability of self-renewal may suggest that such cells are important in the development of lung cancer. The evidence supporting the hypothesis along with its implications are discussed. Confirmation of the hypothesis would suggest that the transition time from a normal cell to overt cancer cell may be much shorter than that based on the multi-step cancer progression model. Additionally, if wounding in other organs is a beacon that attracts bone marrow derived tumor cells, efforts to ameliorate areas of epithelial injury and compensatory wounding may block bone marrow derived tumor cell homing, aberrant repair, and metastasis. Finally, a bone marrow derived lung cancer stem cell would require stem cell poisons for cure. PMID- 16714091 TI - Chemical composition and antimicrobial activity of essential oil from the rhizomes of Amomum cannicarpum. AB - The essential oil obtained by hydrodistillation of the rhizomes of Amomum cannicarpum was analyzed by GC and GC/MS. Thirty-four compounds were identified, of which the major components were beta-terpineol (13.4%), beta-pinene (9.4%) and alpha-pinene (6.9%). The essential oil showed significant antimicrobial activity. PMID- 16714092 TI - Phagocytosis and killing of Streptococcus suis by porcine neutrophils. AB - Streptococcus suis serotype 2 is an important swine pathogen responsible for diverse infections, mainly meningitis. Virulence factors and the pathogenesis of infection are not well understood. Neutrophils may play an important role in the pathogenesis of infection given that infiltration by neutrophils and mononuclear cells are frequently observed in lesions caused by S. suis. The objective of this work was to study the interactions between S. suis serotype 2 and porcine neutrophils. Results showed that suilysin is toxic to neutrophils and this could help S. suis evade innate immunity. Moreover, suilysin appears to affect complement-dependent killing by decreasing the opsonization of S. suis and the bactericidal capacity of neutrophils. Our results confirm that capsule polysaccharide protects S. suis against killing and phagocytosis by neutrophils. We also showed that the presence of specific IgG against S. suis serotype 2 promoted killing by neutrophils, indicating that the induction of a strong humoral response is beneficial for clearance of this pathogen. PMID- 16714093 TI - Identification and characterization of RTVP1/GLIPR1-like genes, a novel p53 target gene cluster. AB - Our previous finding of RTVP1 (GLIPR1) as a p53 target gene with tumor suppressor functions prompted us to initiate a genome-wide sequence homology search for RTVP1/GLIPR1-like (GLIPR1L) genes. In this study we report the identification and characterization of a novel p53 target gene cluster that includes human RTVP1 (hRTVP-1) together with two GLIPR1L genes (GLIPR1L1 and GLIPR1L2) on human chromosome 12q21 and mouse Rtvp1 (mRTVP-1 or Glipr1) together with three Glipr1 like (Glipr1l) genes on mouse chromosome 10D1. GLIPR1L1 has two and GLIPR1L2 has five differentially spliced isoforms. Protein homology search revealed that hRTVP 1 gene cluster members share a high degree of identity and homology. GLIPR1L1 is testis-specific, whereas GLIPR1L2 is expressed in different types of tissues, including prostate and bladder. Like hRTVP-1, GLIPR1L1 and GLIPR1L2 are p53 target genes. The similarities of these novel p53 target gene cluster members in protein structure and their association with p53 suggest that these genes may have similar biological functions. PMID- 16714094 TI - ChickGCE: a novel germ cell EST database for studying the early developmental stage in chickens. AB - We established a database to study germ cells during the early developmental stage in the chicken. The ChickGCE database provides integrated expressed sequence tag (EST) data from chicken testis, ovary, embryonic gonads, and primordial germ cells. We gathered data on 10,294 ESTs from approximately 1000 embryonic gonads, and we experimentally determined 10,851 ESTs from primordial germ cells purified from 7955 embryonic gonads by magnetically activated cell sorting. The EST testis and ovary datasets were retrieved from the public database of The Institute for Genomic Research (TIGR). The EST data were clustered and assembled into unique sequences, contigs, and singletons. The ChickGCE database provides functional annotation, identification, and putative embryonic germ-cell-specific novel transcripts based on the Gene Ontology database, as well as statistical analyses of expression patterns and pair-wise comparisons of two types of tissue- and germ-cell-specific alternative splicing events in the chicken. The new database is accessible online and queries can be answered using several search options, including tissue database searches, keywords, clone IDs, expected values, and BLAST search scores. PMID- 16714095 TI - Identification of a unique splice site variant in SLC39A4 in bovine hereditary zinc deficiency, lethal trait A46: An animal model of acrodermatitis enteropathica. AB - Lethal trait A46, also known as bovine hereditary zinc deficiency, Adema disease, and hereditary parakeratosis, is an autosomal recessive disorder first described in 1964, with a clinical presentation similar to that of acrodermatitis enteropathica (AE) in humans. The molecular basis of the defect has not been previously identified. Recently, the basic defect in AE was found to lie in SLC39A4. We report the characterization of the bovine ortholog of SLC39A4 and identification of a unique splice site variant within this gene in affected animals. The mutation leads to exon skipping, leaving the coding region in frame. The gene product is predicted to lack two critical motifs, which lie in adjacent transmembrane domains implicated in the formation of a pore responsible for the transport of zinc. While further functional studies are warranted, this unique variant is likely to be responsible for the impaired zinc absorption in this disease. PMID- 16714096 TI - Sleeping Beauty transposition in the mouse genome is associated with changes in DNA methylation at the site of insertion. AB - The Sleeping Beauty (SB) transposon (Tn) system is a nonviral gene delivery tool that has widespread application for transfer of therapeutic genes into the mammalian genome. To determine its utility as a gene delivery system, it was important to assess the epigenetic modifications associated with SB insertion into the genome and potential inactivation of the transgene. This study investigated the DNA methylation pattern of an SB Tn as well as the flanking genomic region at insertion sites in the mouse genome. The ubiquitous ROSA26 promoter and an initial part of the eGFP coding sequence in the SB Tn exhibited high levels of CpG methylation in transgenic mouse lines, irrespective of the chromosomal loci of the insertion sites. In contrast, no detectable CpG methylation in the endogenous mouse ROSA26 counterpart was observed in the same animals. Furthermore, significant hypomethylation was detected in neighboring chromosomal sequences of two unique SB Tn insertions compared to wild-type patterns. Taken together, these results suggest that SB Tn insertions into the mouse genome can be discriminated by DNA methylation machinery from an identical endogenous DNA sequence and can profoundly alter the DNA methylation status of the transgene cargo as well as flanking host genomic regions. PMID- 16714097 TI - Altered distribution of leukocyte subsets and cytokine production in response to acute psychosocial stress in patients with psoriasis vulgaris. AB - Psoriasis (PSO) is a mainly T helper-type 1 (TH(1)) cell mediated chronic inflammatory skin disease characterized by epidermal hyperproliferation and psoriatic plaques. There is ample evidence that stress may trigger psoriatic eruption, however, the underlying mechanisms of stress-induced exacerbation of PSO are poorly understood. The specific goal of the present study was to investigate the impact of acute stress on pathologically relevant immune functions in PSO patients. PSO patients (n=23) and healthy controls (n=25) were exposed to a standardized laboratory stressor ("Trier Social Stress Test", TSST) including a free speech and mental arithmetics in front of an audience. Blood samples were collected 10min before and 1, 10, 20, and 60min after the TSST as well as 24h after the experiment at identical time points under resting conditions. Analyses of leukocyte subsets indicated a significantly increased number of leukocyte subpopulations (lymphocytes, granulocytes, CD3(+), CD8(+), CD16(+)/CD56(+), and CD3(+)/HLA-DR(+)) after the TSST (all p<.01) with no significant between-group differences. However, monocyte number (F(3,120)=2.7; p<.01) and number of CD4(+)cells (F(3,120)=3.09; p<.05) were found to be significantly higher in PSO sufferers than in controls. Moreover, a significant decrease of CD3(+)/CD25(+)cells was observed in the PSO, but not in the control group (F(3,120)=3.46; p<.05). After exposure to the TSST, stimulation of peripheral blood mononuclear cells (PBMCs) with phytohemagglutinin (PHA) resulted in elevated production of IFN-gamma (F(3,126)=6.9; p<.001) and IL-2 (F(3,123)=6.6; p<.001), and moreover, a decreased production of IL-10 (F(3,132)=5.22; p<.01) and IL-4 (F(3,129)=3.9; p<.01). No difference in stress induced changes of cytokine production to PHA could be identified between the two experimental groups (all p>.05). The present findings suggest that acute psychosocial stress is associated with changes of immune functions known to be involved in PSO which may be one potential explanation of how stress may trigger psoriatic eruption. PMID- 16714098 TI - Meiotic chromosome abnormalities in human spermatogenesis. AB - The last few years have witnessed an explosion in the information about chromosome abnormalities in human sperm and the meiotic events that predispose to these abnormalities. We have determined that all chromosomes are susceptible to nondisjunction, but chromosomes 21 and 22 and, especially, the sex chromosomes have an increased frequency of aneuploidy. Studies are just beginning on the effects of potential mutagens on the chromosomal constitution of human sperm. The effects of pesticides and cancer therapeutic agents have been reviewed. In the last decade, there has been a great impetus to study chromosome abnormalities in sperm from infertile men because the advent of intracytoplasmic sperm injection (ICSI) made it possible for these men to father pregnancies. A large number of studies have demonstrated that infertile men have an increased frequency of chromosomally abnormal sperm and children, even when they have a normal somatic karyotype. Meiotic studies on the pachytene stage of spermatogenesis have demonstrated that infertile men have impaired chromosome synapsis, a significantly decreased frequency of recombination, and an increased frequency of chromosomes completely lacking a recombination site. Such errors make these cells susceptible to meiotic arrest and the production of aneuploid gametes. PMID- 16714099 TI - Female-specific DNA sequences in ostriches. AB - Ostrich absence of heteromorphic sex chromosomes, unique sequences or markers located in the ostrich W-chromosome. Random amplified polymorphic DNA (RAPD) fingerprinting was carried out to investigate the sex-specific DNA sequence for sexing in ostrich. One hundred and forty random primers were used for random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). One of these primers, OPAJ-13, produced a sex-specific band only found in tested females, which was isolated and constructed into plasmids for nucleotide sequencing. A 760bp novel female-specific sequence was obtained. Two primers (OstSexOPAJ13-F and -R) were designed according to the cloned female sequence to amplify the female-specific fragment from genomic DNA of ostriches for sexing by PCR. The sex specific band was represented in females but none were found in the males. This result showed that the sex of ostrich could be easily and effectively identified using the female-specific primers for PCR technique. PMID- 16714100 TI - Adsorption of cadmium on husk of Lathyrus sativus: physico-chemical study. AB - Adsorption of cadmium (II) from aqueous solution by low-cost biosorbents was investigated. Husk of Lathyrus sativus (HLS) was found to be the most efficient in this respect and removed approximately 95% of the metal. The influence of pH, temperature, contact time and metal ion concentration on the adsorption process by HLS was studied. Hydrogen ion concentration of the solution greatly influenced the process with an optimum at pH 5.0-6.0, whereas temperature had no significant effect. The process was very fast and more than 90% of the total adsorption took place within the first 5 min and was found to follow pseudo-second order rate kinetics. The adsorption data can better be explained by Langmuir isotherm model and the calculated maximum adsorption capacity was 35 mg/g of HLS at pH 5.0 and 30 degrees C. Scanning electron micrographs showed that cadmium was present as micro precipitate on the surface of the adsorbent. Cadmium replaced calcium of the biomass as revealed from the EDX analysis indicating that the adsorption proceeds through ion exchange mechanism. Cadmium could be desorbed from the loaded biomass by lowering pH approximately 1.0 with mineral acid. PMID- 16714102 TI - Characterisation of sugar cane straw waste as pozzolanic material for construction: calcining temperature and kinetic parameters. AB - This paper reports on the influence of calcining temperature (800 and 1000 degrees C) on the pozzolanic activation of sugar cane straw (SCS). The reaction kinetics of SCS ash-lime mixtures were inferred from physicochemical characteristics (X-ray diffraction patterns and thermogravimetry analysis. The fitting of a kinetic-diffusive model to the experimental data (fixed lime versus time) allowed the computing of the kinetic parameters (reaction rate constant) of the pozzolanic reaction. Results obtained confirm that the sugar cane straw ash (SCSA) calcined at 800 and 1000 degrees C have properties indicative of very high pozzolanic activity. No influence of calcining temperature on the pozzolanic activity was observed. Also, no crystalline compounds during the pozzolanic reaction were identified up to 90 days of reaction. Environmental durability and strength of the consequential mortars remain to be assessed. PMID- 16714103 TI - Emissions of organo-metal compounds via the leachate and gas pathway from two differently pre-treated municipal waste materials -- A landfill reactor study. AB - Due to their broad industrial production and use as PVC-stabilisers, agro chemicals and anti-fouling agents, organo-metal compounds are widely distributed throughout the terrestrial and marine biogeosphere. Here, we focused on the emission dynamics of various organo-metal compounds (e.g., di,- tri-, tetra methyl tin, di-methyl mercury, tetra-methyl lead) from two different kinds of pre treated mass waste, namely mechanically-biologically pre-treated municipal solid waste (MBP MSW) and municipal waste incineration ash (MWIA). In landfill simulation reactors, the emission of the organo-metal compounds via the leachate and gas pathway was observed over a period of 5 months simulating different environmental conditions (anaerobic with underlying soil layer/aerated/anaerobic). Both waste materials differ significantly in their initial amounts of organo-metal compounds and their environmental behaviour with regard to the accumulation and depletion rates within the solid material during incubation. For tri-methyl tin, the highest release rates in leachates were found in the incineration ash treatments, where anaerobic conditions in combination with underlying soil material significantly promoted its formation. Concerning the gas pathway, anaerobic conditions considerably favour the emission of organo metal compounds (tetra-methyl tin, di-methyl mercury, tetra-methyl lead) in both the MBP material and especially in the incineration ash. PMID- 16714105 TI - DNA looping: the consequences and its control. AB - The formation of DNA loops by proteins and protein complexes is ubiquitous to many fundamental cellular processes, including transcription, recombination and replication. Recently, advances have been made in understanding the properties of DNA looping in its natural context and how they propagate to cellular behavior through gene regulation. The result of connecting the molecular properties of DNA looping with cellular physiology measurements indicates that looping of DNA in vivo is much more complex and easier than predicted from current models, and reveals a wealth of previously unappreciated details. PMID- 16714104 TI - DNA architecture: from G to Z. AB - G-quadruplexes and Z-DNA are two important non-B forms of DNA architecture. Results on novel structural elements, folding and unfolding kinetics, and interactions with small molecules and proteins have been reported recently for these forms. These results will enhance our understanding of the biology of these structures and provide a platform for drug design. PMID- 16714106 TI - Structure of the '30 nm' chromatin fibre: a key role for the linker histone. AB - The structure of the '30 nm' chromatin fibre has eluded us for 30 years and remains a major unsolved problem in biology. Progress during the past year has led to the proposal of two significantly different models: one derived from the crystal structure of a four-nucleosome core array lacking the linker histone and the other, much more compact structure, derived from electron microscopy analysis of long nucleosome arrays containing the linker histone. The first model is of the two-start helix type, the second a one-start helix with interdigitated nucleosomes. These models provide new evidence that the topology and compactness of the '30 nm' chromatin fibre structure are regulated by the linker histone. The structural information also provides insights into the mechanisms by which the degree of chromatin compaction might be regulated by histone composition and post transcriptional modifications. PMID- 16714107 TI - Effect of refining on pulp surface charge accessible to polydadmac and FTIR characteristic bands of high yield kraft fibres. AB - Studies were carried out to determine the surface charge and total charge of refined softwood high yield kraft pulps at different levels by titration methods. The results showed that the surface charge was highly dependent on the degree of refining. However, the total fibre charge was not affected by refining. The change of the fibre surface charge during refining monitored using ATR-FTIR characteristic bands was within 1700-1300 cm(-1). Reliable PLS1 calibration models could be established to correlate the FTIR spectral data and the surface charge of the refined pulps. PMID- 16714108 TI - Identification of thieno[3,2-b]pyrroles as allosteric inhibitors of hepatitis C virus NS5B polymerase. AB - Thieno[3,2-b]pyrroles are a novel class of allosteric inhibitors of HCV NS5B RNA dependent RNA polymerase which show potent affinity for the NS5B enzyme. Introduction of a polar substituent in the position N1 led to a compound that efficiently blocks subgenomic HCV RNA replication in HUH-7 cells with an EC50 of 2.9 microM. PMID- 16714109 TI - Design of novel, potent, and selective human beta-tryptase inhibitors based on alpha-keto-[1,2,4]-oxadiazoles. AB - A series of novel alpha-keto-[1,2,4]-oxadiazoles has been synthesized as human tryptase inhibitors for evaluation as a new class of anti-asthmatic agent. The inhibitor design is focused on using a prime-side hydrophobic pocket and the S2 pocket of beta-tryptase to achieve inhibition potency and selectivity over other serine proteases. PMID- 16714110 TI - Synthesis and biological activity of nociceptin/orphanin FQ(1-13)NH2 analogues modified in 9 and/or 13 position. AB - The purpose of the present study was the synthesis and the biological screening of new analogues of N/OFQ(1-13)NH2, the minimal sequence maintaining the same activity as the natural peptide nociceptin. In order to investigate the role of Lys, we substituted Lys at positions 9 and/or 13 by Orn, Dab (diaminobutanoic acid) or Dap (diaminopropanoic acid). The new N/OFQ(1-13)NH2 analogues exerted strong and naloxone-resistant inhibition of electrically evoked contractions of rat vas deferens. Lys replacement with Orn maintained or even enhanced the inhibitory activity, while replacements with Dab and Dap decreased inhibitory activity. PMID- 16714111 TI - Lipase catalyzed synthesis of benzyl acetate in solvent-free medium using vinyl acetate as acyl donor. AB - Use of vinyl acetate as acyl donor in transesterification of benzyl alcohol catalyzed by a commercially available lipase (Lipozyme RM IM) gave 100% conversion in 10 min. The excess acyl donor and the enzyme could be recovered and reused. Unlike the chemical catalytic processes, it produced no undesirable side product. PMID- 16714112 TI - Estimation of the hydrophobicity of 2,4-diphenyl-1,3-oxazoline analogs and QSAR analysis of their ovicidal activity against Tetranychus [corrected] urticae. AB - Partition coefficients of six 2-phenyl-1,3-oxazoline congeners containing 2-I, 2 NO2, 2-CF3, 2,6-(CH3)2, 2,6-F2, and 2-F-6-Cl substitutions on the phenyl moiety were measured in a 1-octanol/water system using the flask-shaking method. The effect on the hydrophobicity (LogP) of substituents on the phenyl moiety of 2 phenyl-1,3-oxazolines linearly correlated with that of benzamide congeners. logP values of other 2-(substituted phenyl)-1,3-oxazoline analogs were empirically estimated from the corresponding substituted benzamides. The ovicidal activity of 2-(substituted phenyl)-4-phenyl-1,3-oxazoline analogs against the two-spotted spider mite Tetranychus [corrected] urticae was quantitatively analyzed using the classical QSAR (Hansch-Fujita) method. Results showed that ovicidal activity increases with hydrophobicity. The introduction of inductive electron-withdrawing groups at ortho-positions increased ovicidal activity, but addition of steric bulk was unfavorable. Substitution at either the meta- or para-position was detrimental to the acaricidal activity. PMID- 16714113 TI - The effects of muscle damage following eccentric exercise on gait biomechanics. AB - To examine the effects of knee extensors muscle damage on walking and running biomechanics in healthy males. Muscle damage was caused by 60 (6x10) maximal eccentric knee flexions of both legs, selected in a random order, at an angular velocity of 1.05rad/s in 10 volunteers (mean age 20+/-1.0 years). Muscle damage indicators (creatine kinase (CK), lactate dehydrogenase (LDH), delayed onset muscle soreness (DOMS), eccentric and isometric (110 degrees knee flexion) peak torque), pelvic three dimensional (3D) orientation, as well as hip, knee and ankle-joint flexion/extension angles during gait (walking at 1.2m/s and running at 2.8m/s) were assessed pre- and 48h post-eccentric exercise. All muscle damage indicators revealed significant changes post- compared to pre-exercise data (P<0.05) confirming that muscle damage did occur. Kinematic analysis revealed that muscle damage significantly decreased the knee-joint angle range of movement at the stance and swing phases during walking (P<0.05) and running (P<0.05), respectively. These changes were accompanied by corresponding increases of pelvic rotation (P<0.05) and decrease of pelvic tilt (P<0.05). The present data demonstrate that damage of knee extensors result in changes of treadmill walking and running kinematics at both knee joint and pelvis. The fact that these alterations occur at different gait phases could be attributed to the speed of movement and to a self-protection mechanism to prevent further damage. PMID- 16714114 TI - QSAR analysis of antimicrobial and haemolytic effects of cyclic cationic antimicrobial peptides derived from protegrin-1. AB - In this paper we quantitatively analyse antimicrobial and haemolytic activities of porcine protegrin-1 (PG-1) mimetics-cyclic cationic peptides with beta-hairpin fold synthesised by Robinson et al. [Bioorg. Med. Chem.2005, 13, 2055]. The presented QSAR models, which use molecular properties related to possible mechanisms of cell membrane disruption that can be easily calculated from available data on amino acids, rationalize the relationships between sequences and antimicrobial and haemolytic potencies of the cyclic peptides. The best models obtained by application of genetic function approximation algorithm correlate antimicrobial potencies to the peptide's charge and amphipathicity index, while the haemolytic effect correlates well with the lipophilicity of residues forming the nonpolar face of the beta-hairpin. The models permit selection of site-directed residue substitutions leading to simultaneous optimization of antimicrobial and haemolytic potencies. Examples of such residue substitutions in the nonpolar face of a symmetric cyclic beta-hairpin PG-1 analogue with an ideal amphipathic structure are given. PMID- 16714115 TI - Caulerpenyne-colchicine hybrid: synthesis and biological evaluation. AB - The synthesis of an analog of caulerpenyne having a trimethoxyaryl moiety was achieved in 11% overall yield over 11 steps. Its biological activity has been evaluated as inhibitor of in vitro tubulin polymerization or angiogenesis. PMID- 16714116 TI - Synthesis and anticancer evaluation of bis(benzimidazoles), bis(benzoxazoles), and benzothiazoles. AB - Four classes of UK-1 analogues were synthesized and their cytotoxicity testing against human A-549, BFTC-905, RD, MES-SA, and HeLa carcinoma cell lines was determined. The results revealed that UK-1 and four of these analogues (15-18) are potent against the cancer cell lines. In particular, compound 16 is more potent than UK-1 against A-549 and HeLa cell lines, and compounds 15, 17, and 18 selectively exhibit potent cytotoxic activity against the BFTV-905 cells (IC50 9.6 microM), A-549 cells (IC50 6.6 microM), and MES-SA cells (IC50 9.2 microM), respectively. PMID- 16714117 TI - Luminescent pyrimidine hydrazide oligomers with peptide affinity. AB - The modular synthesis of pyrimidine oligohydrazides and their peptide binding ability are reported. Ethylene glycol substituents ensure water solubility of the compounds. The pattern of hydrogen bond donors and hydrogen bond acceptors resembles the functionalities of a peptide backbone, and intramolecular hydrogen bonds restrict conformational mobility. The pyrimidine heterocycles show emission at 423 nm if either excited with light of 320 nm or by a FRET process from a nearby Trp residue. This property is useful for the luminescent detection of interactions with peptides and proteins. PMID- 16714118 TI - Practical aspects of electron energy-loss spectroscopy (EELS) calculations using FEFF8. AB - We discuss the application of the ab initio program FEFF8 to calculations of electron energy-loss spectroscopy (EELS), focusing in particular on core-loss spectra. FEFF8 is based on a self-consistent, real space multiple scattering formalism. We focus on issues relevant to practical simulations, including the construction of well-converged potentials, the treatment of inelastic losses and exchange-correlation potentials and the core-hole. We also discuss how to account for experimental conditions, for example, sample orientation and finite temperature effects such as Debye-Waller factors. Finally we discuss the interpretation of the spectra in terms of electronic structure and local projected density of states (LDOS). As an explicit example, we illustrate various features of the code by application to the ionization edges of GaN. PMID- 16714119 TI - Self-aggregation of fibrillar collagens I and II involves lysine side chains. AB - Several properties of fibrillar collagens depend on abundance and position of ionic amino acids. We recently demonstrated that N-methylation and N-acetylation of Lys/Hyl amino group did not significantly alter the thermal stability of the triple helical conformation and that the binding of modified collagens I and II to decorin is lost only on N-acetylation. The positive charge at physiological pH of Lys/Hyl side chains is preserved only by N-methylation. We report here the new aspect of the influence of the same modifications on collagen self-aggregation in neutral conditions. Three collagen preparations are very differently affected by N-methylation: acid-soluble type I collagen maintains the ability to form banded fibrils with 67-nm periodicity, whereas almost no structured aggregates were detected for pepsin-soluble type I collagen; pepsin-soluble type II collagen forms a very different supramolecular species, known as segment long spacing (SLS). N-acetylation blocks the formation of banded fibrils in neutral conditions (as did all other chemical modifications reported in the literature), demonstrating that the positive charge of Lys/Hyl amino groups is essential for self-aggregation. Kinetic measurements by turbidimetry showed a sizeable increase of absorbance only for the two N-methylated samples forming specific supramolecular aggregates; however, the derivatization affects aggregation kinetics by increasing lag time and decreasing maximum slope of absorbance variation, and lowers aggregation competency. We discuss that the effects of N methylation on self-aggregation are caused by fewer or weaker salt bridges and by decrease of hydrogen bonding potential and conclude that protonated Lys side chains are involved in the fibril formation process. PMID- 16714120 TI - Isoquino[4,5-bc]acridines: design, synthesis and evaluation of DNA binding, anti tumor and DNA photo-damaging ability. AB - Several novel isoquino[4,5-bc]acridine derivatives have been designed and synthesized. Their DNA-binding, anti-tumor and DNA-photo-damaging properties were investigated. A4 exhibited the highest anti-tumor activities against both A 549 (human lung cancer cell) and P388 (murine leukemia cells). All these compounds were found to be more cytotoxic against P388 than against A549. Under 365-nm light irradiation, A3 damaged plasmid DNA pBR322 at <2 microM and cleaved DNA from form I to 100% form II by 50 microM. The mechanism studies revealed that A3 damaged DNA by electron transfer mechanism and singlet oxygen species. PMID- 16714121 TI - The metabolic syndrome. Beyond the insulin resistance syndrome. PMID- 16714122 TI - Nanomechanical properties of desmin intermediate filaments. AB - Desmin intermediate filaments play important role in the mechanical integrity and elasticity of muscle cells. The mechanisms of how desmin contributes to cellular mechanics are little understood. Here, we explored the nanomechanics of desmin by manipulating individual filaments with atomic force microscopy. In complex, hierarchical force responses we identified recurring features which likely correspond to distinct properties and structural transitions related to desmin's extensibility and elasticity. The most frequently observed feature is an initial unbinding transition that corresponds to the removal of approximately 45-nm-long coiled-coil dimers from the filament surface with 20-60 pN forces in usually two discrete steps. In tethers longer than 60 nm we most often observed force plateaus studded with bumps spaced approximately 16 nm apart, which are likely caused by a combination of protofilament unzipping, dimer-dimer sliding and coiled-coil-domain unfolding events. At high stresses and strains non-linear, entropic elasticity was dominant, and sometimes repetitive sawtooth force transitions were seen which might arise because of slippage within the desmin protofilament. A model is proposed in which mechanical yielding is caused by coiled-coil domain unfolding and dimer-dimer sliding/slippage, and strain hardening by the entropic elasticity of partially unfolded protofilaments. PMID- 16714123 TI - Sex differences in mental rotation: top-down versus bottom-up processing. AB - Functional MRI during performance of a validated mental rotation task was used to assess a neurobiological basis for sex differences in visuospatial processing. Between-sex group analysis demonstrated greater activity in women than in men in dorsalmedial prefrontal and other high-order heteromodal association cortices, suggesting women performed mental rotation in an effortful, "top-down" fashion. In contrast, men activated primary sensory cortices as well as regions involved in implicit learning (basal ganglia) and mental imagery (precuneus), consistent with a more automatic, "bottom-up" strategy. Functional connectivity analysis in association with a measure of behavioral performance showed that, in men (but not women), accurate performance was associated with deactivation of parieto-insular vestibular cortex (PIVC) as part of a visual-vestibular network. Automatic evocation by men to a greater extent than women of this network during mental rotation may represent an effective, unconscious, bottom-up neural strategy which could reasonably account for men's traditional visuospatial performance advantage. PMID- 16714124 TI - HIV-1 protein gp120 rapidly impairs memory in chicks by interrupting the glutamate-glutamine cycle. AB - Learning and memory impairments are frequently observed in patients suffering from AIDS Dementia Complex (ADC). These effects have been linked to the presence of gp120, an HIV viral coat glycoprotein. The present study investigated the possibility that gp120 prevents the uptake of extracellular glutamate by astrocytes, leading to an interruption of the glutamate-glutamine cycle and a subsequent impairment of memory. Ten microliters of 10nM gp120 was bilaterally injected into the region of the intermediate medial mesopallium of day-old chicks at various times before, or after, training using a single-trial passive avoidance task. Gp120 was found to significantly impair memory retention when injected 10-40 min after training. Memory impairments were evident within 5 min of gp120 administration and remained evident 24h later. Further, the amnestic effect of gp120 could be overcome with glutamine or with precursors of glutamate synthesis, but only weakly by glutamate. These results support the conclusion that the amnestic effect of gp120 is due to an impaired uptake of glutamate by astrocytes and a subsequent interruption of glutamine supply to neurones. The data indicate that the glutamate-glutamine cycle may be a useful therapeutic target in the treatment of ADC. PMID- 16714125 TI - Excitotoxic lesions of the parafascicular nucleus produce deficits in a socially transmitted food preference. AB - The parafascicular (PF) nucleus, a posterior component of the intralaminar nuclei of the thalamus, is considered to be an essential structure in the feedback circuits of basal ganglia-thalamo-cortical systems that critically participate in cognitive processes. To study the PF contribution to processing of behaviorally significant information during specific episodes of learning, we investigated the effects of damaging the PF nucleus in the acquisition of a natural form of social olfactory learning, the socially transmitted food preference (STFP) task. This task is a non-spatial paradigm that exhibits some of the characteristics of relational memory because it requires that animals use information obtained in one episode to guide later behavior in different circumstances. Adult male Wistar rats were submitted to pretraining bilateral N-methyl-D-aspartate (0.15 M, pH 7.4) lesions of the PF (0.4 microl/side, 0.2 microl/min). The behavioral effects of PF lesions were compared to vehicle- and sham-operated control groups and two retention delays were considered in separate groups: immediately (Lesion-I, Vehicle-I, and Sham-I groups) and 24h after training (Lesion-24, Vehicle-24, and Sham-24 groups). PF lesions produced delay-independent impairments in the STFP suggesting that this nucleus might modulate the acquisition of this odor-odor association task. Results are discussed in the context of medial prefrontal cortex deafferentation induced by PF damage. PMID- 16714126 TI - The canonical Wnt pathway in embryonic axis polarity. AB - The canonical Wnt pathway plays crucial roles in multiple developmental processes, including in axis specification. Throughout the animal kingdom, this pathway has been reported to drive patterning of axes as different as the animal vegetal axis in echinoderms to the dorsal-ventral axis in vertebrates. Intriguingly enough, this pathway appears structurally and functionally well conserved during evolution. However, differences between these phyla are observed that explain how a same pathway can mediate establishment of two such apparently distinct axes. This review compares the axis specification processes used in two evolutionarily distant embryos, the sea urchin and Xenopus. PMID- 16714128 TI - Sensitivity to local dipole fields in the CRAZED experiment: an approach to bright spot MRI. AB - Local dipole fields such as those created by small iron-oxide particles are used to produce regions of low intensity (dark contrast) in many molecular magnetic resonance imaging applications. We have investigated, with computer simulations and experiments at 17.6 T, how the COSY revamped with asymmetric z-gradient echo detection (CRAZED) experiment that selects intermolecular double-quantum coherences can also be used to visualize such local dipole fields. Application of the coherence-selection gradient pulses parallel to the main magnetic field produced similar, dark contrast as conventional gradient echo imaging. Application of the gradient along the magic angle leads to total loss of signal intensity in homogeneous samples. In the presence of local dipole fields, the contrast was inverted and bright signals from the dipoles were observed over a very low background. Both simulations and experiments showed that the signal strongly decreased when a phase-cycle suppressing single-quantum coherences was employed. Therefore, we conclude that most of the signal comes from directly refocused magnetization or intermolecular single-quantum coherences. Finally, we demonstrate that bright contrast from local dipole fields can also be obtained, when the pair of coherence-selection gradient pulses is deliberately mismatched. Both methods allowed visualization of local dipole fields in phantoms in experimental times of about 3 min. PMID- 16714127 TI - Improvements to gene deletion in the fungal pathogen Cryptococcus neoformans: absence of Ku proteins increases homologous recombination, and co-transformation of independent DNA molecules allows rapid complementation of deletion phenotypes. AB - Cryptococcus neoformans is a pathogenic fungus that is relatively amenable to molecular genetic analysis, including gene deletion. However, rates of homologous recombination can be low, so obtaining specific gene deletion transformants is challenging. We have utilized two new technologies, cku deletion strains to improve the efficiency of gene deletions in this organism, and co transformations. The Ku70-Ku80 heterodimer is predicted to be an essential part of the non-homologous end-joining process in C. neoformans. Here we show that a deletion in one or both of these proteins results in an increase in the rates of homologous recombination. Importantly, we demonstrate that after generation of a strain with a particular deletion of interest, the cku deletion can be removed by mating and segregation. We also utilize co-transformation of wild-type genes and selectable markers on separate linear DNA molecules to complement a deletion event. We show that co-transformation results in the successful restoration of wild-type phenotype, though variations in this phenotype often occur. PMID- 16714129 TI - HAT HMBC: a hybrid of H2BC and HMBC overcoming shortcomings of both. AB - A new 2D NMR experiment, HAT HMBC, that is a hybrid of H2BC and HMBC aims at establishing two-bond correlations absent in H2BC spectra because of vanishing (3)J(HH) coupling constants. The basic idea is to create an additional pi phase difference in the multiplet structure in HMBC peaks with respect to the (n+1)J(HH) coupling constant between the proton(s) attached to a (13)C and a (1)H separated by n bonds. Thus HMBC peaks associated with small J(HH) will be the most attenuated in a HAT HMBC spectrum in comparison to a regular HMBC spectrum, i.e. peaks associated with (n+1)J(HH) and (n)J(CH) will for n>2 usually be strongly attenuated. The HAT HMBC pulse sequences contain the same number of pulses as regular HMBC and are only a few milliseconds longer. PMID- 16714130 TI - Development of maximum metabolic rate and pulmonary diffusing capacity in the superprecocial Australian Brush Turkey Alectura lathami: an allometric and morphometric study. AB - The Australian Brush Turkey Alectura lathami is a member of the Megapodiidae, the mound-building birds that produce totally independent, "superprecocial" hatchlings. This study examined the post-hatching development of resting and maximal metabolic rates, and the morphometrically determined changes in pulmonary gas exchange anatomy, in chicks during 3.7 months of growth from hatchlings (122 g) to subadults (1.1 kg). Allometric equations of the form y=aM(b) related gas exchange variables (y) to body mass (M, g). Metabolic rates were measured with open-flow respirometry (mL O2 min(-1)) of chicks resting in the dark and running above the aerobic limit on a treadmill. Resting metabolic rate (RMR=0.02 M(0.99)) and maximal metabolic rate (MMR=0.05 M(1.07)) scaled with exponents significantly above those of interspecific allometries of adult birds. However MMR was below that expected for other species of adult birds in flapping flight, consistent with the Brush Turkey's ground-dwelling habits. Total lung volumes (mL) increased faster than isometrically (V(L)=0.0075 M(1.19)), as did the surface area (cm(2)) of the blood-gas barrier (S(t)=7.80 M(1.23)), but the data overlapped those of adult species. Harmonic mean thickness of the blood-gas barrier was independent of body size (mean tau(ht),=0.39 microm) and was about twice that expected for flying birds. Diffusing capacity (mL O2 min(-1) kPa(-1)) of the blood-gas tissue barrier increased faster than isometrically (Dto2=0.049 M(1.23)); in hatchling Brush Turkeys, it was about 30% expected for adult birds, but this difference disappeared when they became subadults. When compared to altricial Australian pelicans that hatch at similar body masses, superprecocial Brush Turkeys had higher MMR and higher Dto2 at the same body size. A parallel allometry between MMR and Dto2 in Brush Turkeys and pelicans is consistent with the concept of symmorphosis during development. PMID- 16714131 TI - Accelerated recovery of Atlantic salmon (Salmo salar) from effects of crowding by swimming. AB - The effects of post-crowding swimming velocity (0, 0.35, and 0.70 m/s) and recovery time (1.5, 6, and 12 h) on physiological recovery and processing quality parameters of adult Atlantic salmon (Salmo salar) were determined. Atlantic salmon crowded to a density similar to that of a commercial slaughter process (>200 kg/m(3), 40 min) were transferred to a swimming chamber for recovery treatment. Osmolality and concentrations of cortisol, glucose and lactate in blood plasma were used as physiological stress indicators, whereas image analyses of extent and duration of rigor contraction, and fillet gaping were used as measures of processing quality. Crowded salmon had a 5.8-fold higher plasma cortisol concentration than control salmon (P<0.05). The elevated plasma cortisol concentration was reduced by increasing the swimming velocity, and had returned to control levels after 6 h recovery at high water velocity. Similar effects of swimming velocity were observed for plasma osmolality and lactate concentration. A lower plasma glucose concentration was present in crowded than in control fish (P<0.05), although a typical post-stress elevation in plasma glucose was observed after the recovery treatments. Lower muscle pH was found in crowded compared with control salmon (P<0.05), but muscle pH returned to control levels after 6 h recovery at intermediate and high swimming velocities and after 12 h in the low velocity group. Crowding caused an early onset of rigor mortis contraction. However, subjecting crowded salmon to active swimming for 6 h before slaughter delayed the onset of rigor mortis contraction from 2.5 to 7.5 h post mortem. The extent of rigor mortis contraction was also affected by crowding and post-stress swimming activity (P<0.05), and the largest degree of contraction was found in crowded salmon. In conclusion, active swimming accelerated the return of plasma cortisol, hydromineral balance, and the energy metabolism of adult Atlantic salmon to pre-stress levels. Moreover, an active swimming period delayed the onset of rigor mortis contraction, which has a positive technological implication for the salmon processing industry. PMID- 16714132 TI - Bariatric surgery for morbidly obese adolescents: is there a rationale for early intervention? AB - Obesity is one of the most common metabolic disorders affecting the US population: 31% of adults and 16% of adolescents now meet the criteria for obesity. Similar to the rising prevalence of morbid obesity in adults, the prevalence of more severe obesity is also increasing in pediatric patients. The most effective treatment for morbid obesity is bariatric surgery, a procedure most commonly performed in the fifth and sixth decades of life. Although it is clear that rapid, profound weight loss can significantly improve multiple comorbid conditions in adults, including disordered carbohydrate metabolism, obstructive sleep apnea, and cardiovascular risk factors, it is not clear to what degree similar comorbidities are affected in adolescents undergoing surgical weight loss. In this paper, the indications, contraindications, and early surgical outcomes of gastric bypass surgery for morbidly obese adolescents are reviewed, and important directions for future research are discussed. PMID- 16714134 TI - Salmonella resistant to extended-spectrum cephalosporins: prevalence and epidemiology. AB - Salmonella resistant to extended-spectrum cephalosporins (ESCs) have emerged worldwide since 1988. By 2004, 43 countries had reported this public health problem. Resistance was mediated by classical extended-spectrum beta-lactamases, plasmid-mediated cephalosporinases, and recently a class A carbapenemase. Of these, CMY-2 is the most widely disseminated enzyme. Salmonella enterica serotype Typhimurium and S. enterica serotype Enteritidis are the most common serovars associated with ESC resistance in human infections. Many outbreaks in humans have been reported, most often among children and neonates. ESC-resistant Salmonella is frequently recovered from animals and food, with poultry as primary food source, suggesting that humans are often infected by these routes. PMID- 16714133 TI - Combined methylmalonic aciduria and homocystinuria (cblC): phenotype-genotype correlations and ethnic-specific observations. AB - Methylmalonic aciduria and homocystinuria, cblC type (MIM 277400), is the most frequent inborn error of vitamin B12 (cobalamin, Cbl) metabolism, caused by an inability of the cell to convert Cbl to both of its active forms (MeCbl, AdoCbl). Although considered a disease of infancy, some patients develop symptoms in childhood, adolescence, or adulthood. The gene responsible for cblC, MMACHC, was recently identified. We studied phenotype-genotype correlations in 37 patients from published case-reports, representing most of the landmark descriptions of this disease. 25/37 had early-onset disease, presenting in the first 6 months of life: 17/25 were found to be either homozygous for the c.271dupA mutation (n=9) or for the c.331C>T mutation (n=3), or compound heterozygotes for these 2 mutations (n=5). 9/12 late-onset cases presented with acute neurological symptoms: 4/9 were homozygous for the c.394C>T mutation, 2/9 were compound heterozygotes for the c.271dupA and c.394C>T mutations, and 3/9, for the c.271dupA mutation and a missense mutation. Several observations on ethnic origins were noted: the c.331C>T mutation is seen in Cajun and French-Canadian patients and the c.394C>T mutation is common in the Asiatic Indian/Pakistani/Middle Eastern populations. The recognition of phenotype genotype correlations and the association of mutations with specific ethnicities will be useful for identification of disease-causing mutations in cblC patients, for carrier detection and prenatal diagnosis in families where mutations are known, and in setting up initial screening programs in molecular diagnostic laboratories. Further study into disease mechanism of specific mutations will help to understand phenotypic presentations and the overall pathogenesis in cblC patients. PMID- 16714135 TI - The clonal spread of multidrug-resistant non-typhi Salmonella serotypes. AB - Non-typhoid Salmonella are one of the most important organisms causing food-borne diseases worldwide. There have been significant increases in developed countries in recent years in the occurrence of resistance, in particular multidrug resistance phenotypes, in non-typhoid Salmonella spp. Such increases have been observed in many countries, not only within the European community but also the Americas and Southeast Asia. Of particular concern is the increasing detection of Salmonella isolates displaying resistance to key antimicrobials, notably fluoroquinolones and third-generation cephalosporins. An important factor associated with this increase in multidrug resistance among particular Salmonella spp. is the national and international spread of certain clonal genotypes, the most recent being the global epidemic spread of multidrug-resistant S. Typhimurium DT104, since the early 1990s. In this review, we describe examples where particular antimicrobial-resistant Salmonella serotypes emerged, persisted for periods of time, and then quickly decreased in prevalence. PMID- 16714136 TI - Introduction: emerging antimicrobial resistance mechanisms in the zoonotic foodborne pathogens Salmonella and Campylobacter. PMID- 16714137 TI - Resistance to fluoroquinolones in Salmonella: emerging mechanisms and resistance prevention strategies. AB - We review the current state of knowledge about the genetic and biochemical mechanisms that mediate quinolone resistance in Salmonella. They include modifications of topoisomerase targets, increased efflux activity and the recently described topoisomerase protection by the plasmid-encoded Qnr protein. We discuss what factors may determine the order of implementation of these various mechanisms in a particular strain, and what strategies could be used to combat resistance, from the inhibition of mutagenesis mechanisms to counteracting, during fluoroquinolone treatment, of resistance mechanisms already set in the infecting strain. PMID- 16714138 TI - Fitness of antimicrobial-resistant Campylobacter and Salmonella. AB - Campylobacter and Salmonella are the most commonly reported bacterial causes of human foodborne infections, and increasing proportions of these pathogens become resistant to medically important antimicrobial agents, imposing a burden on public health. Acquisition of resistance to antibiotics affects the adaptation and evolution of Salmonella and Campylobacter in various environments. Many resistance-conferring mutations entail a biological fitness cost, while others (e.g. fluoroquinolone resistance in Campylobacter) have no cost or even enhanced fitness. In Salmonella, the fitness disadvantage due to antimicrobial resistance can be restored by acquired compensatory mutations, which occur both in vitro and in vivo. The compensated or even enhanced fitness associated with antibiotic resistance may facilitate the spread and persistence of antimicrobial-resistant Salmonella and Campylobacter in the absence of selection pressure, creating a significant barrier for controlling antibiotic-resistant foodborne pathogens. PMID- 16714139 TI - Microtubule-targeting agents in angiogenesis: where do we stand? AB - Angiogenesis is a key event of tumor progression and metastasis and hence a target for cancer chemotherapy. Therapeutic strategies focused on angiogenesis include the discovery of new, targeted anti-angiogenic agents and the re evaluation of conventional anti-cancer drugs. Here, we review the most recent studies investigating the molecular and cellular mechanisms responsible for the anti-angiogenic activity of microtubule-targeting agents (MTAs). These agents include some of the most widely used and effective antitumor drugs that are also among the most anti-angiogenic. In addition, we summarize the latest results of pre-clinical and clinical studies involving MTAs administered at low metronomic doses and in anti-angiogenic combination strategies. Finally, we discuss the future development of these agents, their clinical potential and their limitations. PMID- 16714140 TI - Evolution of parasitic symbioses between plants and filamentous microorganisms. AB - Innate defense in wild plant populations is an invaluable resource for understanding how sustainable disease control can be achieved in crops through research that is rooted in molecular and evolutionary biology. Much progress has been made from molecular research into pathogen detection and defense induction. Bacterial pathology of the wild species Arabidopsis thaliana is at the forefront in revealing parallels with animal innate immunity against infectious diseases. In plants, unlike in animals, however, expansion in biodiversity has been mirrored by tremendous diversification in filamentous parasites. The fungal and oomycete pathology of Arabidopsis is exposing opportunities to investigate the molecular bases of compatibility, plant-driven speciation of parasites, and molecular epidemiology. Such research might reveal evidence that an arms race did occur in the evolution of plant-parasite symbioses. PMID- 16714141 TI - Show and tell: cell biology of pathogen invasion. AB - Because the initial stages of pathogen invasion are often confined to a limited number of host cells, measures of host responses that are averaged over attacked and non-attacked cells provide an unsatisfactory view of these events. To identify the earliest and often transient responses to pathogen attack, there is considerable interest in monitoring the subcellular events that occur specifically in living host cells. Recent improvements in live-cell imaging using fluorescent-tagged markers have expanded the scope of the experiments that can be performed. Changes in the subcellular distribution of organelles and of fluorescently tagged proteins can be monitored in real time in living tissues during pathogen attack, and the dynamic nature of such changes across space and over time can be determined. The application of these sensitive imaging methods has extended earlier observations, made with Nomarski microscopy or inferred from static transmission electron micrographs, about the focal accumulation of subcellular organelles at sites of pathogen attack. In addition, recent experiments have demonstrated the focused accumulation and interaction of specific plant proteins at penetration sites, opening a new window on early host responses and raising questions about the underlying plant processes that sense and direct this marshalling of host resources to block pathogen entry. PMID- 16714142 TI - Preparation and characterization of Nd3+ doped sodium leadbismuthate glass. AB - Nd(3+) doped unconventional sodium leadbismuthate glass is prepared through the melt quenching method. The amorphous nature of the glass is confirmed through the X-ray diffraction study. The differential thermal study was performed to identify the glass transition and approximate glass thermal stability measurements. To identify the local structure of the glass, Fourier transform infrared spectral analysis was also carried out. By performing the Judd-Ofelt theory, the theoretical and experimental oscillator strengths were calculated. PMID- 16714143 TI - Analysis of phospholipid species in rat peritoneal surface layer by liquid chromatography/electrospray ionization ion-trap mass spectrometry. AB - The main phospholipids in rat peritoneal surface layer were analyzed by normal phase high-performance liquid chromatography (HPLC) coupled with electrospray ionization (ESI) ion-trap mass spectrometry (MS). By using a silica gel column and a gradient of hexane/isopropanol/water as mobile phase containing 5 mmol/L ammonium formate as modifiers, a baseline separation of glycerophosphoehtanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidylcholine (PC), sphingomyelin (SM) and lyso-phosphatidylcholine (LPC) was obtained and more than 90 phospholipid constituents in rat peritoneal surface were identified and determined by on-line ion-trap MS detection. The major ethanolamine glycerophospholipids in rat peritoneal surfaces were plasmalogens that were highly enriched in polyunsaturated fatty acids at the sn-2 position. In addition, the fragmentation patterns for each phospholipid class by the ion-trap MS were discussed. PMID- 16714145 TI - 'Frequent' ventricular bigeminy--a reversible cause of dilated cardiomyopathy. How frequent is 'frequent'? AB - An interesting development in the field of heart failure has been the link between frequent premature ventricular contractions and cardiomyopathy. We report a patient whose frequent ventricular bigeminy resulted in left ventricular impairment that resolved after the use of non-contact mapping during radiofrequency ablation. A review of the literature regarding possible mechanisms is discussed. For the practicing clinician, the question of 'frequent' should be taken in context of symptoms and LV function. A single 24-h Holter monitor may not truly reflect the ectopic load. We recommend that if there is associated LV dysfunction and a causal link to frequent PVCs then suppression with radiofrequency ablation is a safe and effective treatment strategy. PMID- 16714144 TI - Elevated circulating levels of heat shock protein 70 are related to systemic inflammatory reaction through monocyte Toll signal in patients with heart failure after acute myocardial infarction. AB - BACKGROUND: Recent studies have shown that heat shock protein (HSP) 70 may serve as a "damage signal" to the immune system and could be the endogenous ligand for Toll-like receptor (TLR) 4 mediating synthesis of inflammatory cytokines. AIMS: To explore the relationship between circulating HSP70 levels and activation of monocyte TLR4 and myocardial damage after AMI. METHODS AND RESULTS: This study examined circulating HSP70 and monocyte TLR4 levels in 52 patients with AMI and 20 controls, and analyzed ex vivo inflammatory cytokine productions using HSP70 stimulated monocytes. Circulating HSP70 levels were higher in AMI patients on day 1 after onset than in controls and remained elevated in AMI patients 14 days after onset. HSP70 levels were positively correlated with monocyte TLR4, plasma interleukin-6 and tumor necrosis factor-alpha levels in AMI patients. HSP70 levels 14 days after onset were higher in AMI patients with heart failure (n=15) than in those without heart failure. In our in vitro study, HSP70-stimulated monocytes resulted in dose-dependent TLR4 expression and release of inflammatory cytokines. TLR4 antibody inhibited inflammatory cytokines release. CONCLUSIONS: Elevated circulating levels of HSP70 may be involved in TLR4 signal-mediated immune response and the progression of heart failure after AMI. PMID- 16714146 TI - Effects of home-based exercise training on neurovascular control in patients with heart failure. AB - BACKGROUND: The effect of home-based exercise training on neurovascular control in heart failure patients is unknown. AIMS: To test the hypothesis that home based training would maintain the reduction in muscle sympathetic nerve activity (MSNA) and forearm vascular resistance (FVR) acquired after supervised training. METHODS AND RESULTS: Twenty-nine patients (54+/-1.9 years, EF<40%) were randomised into two groups: untrained control (n=12) and exercise trained (n=17). Both groups underwent assessment of Quality of Life (QoL), MSNA, and forearm blood flow. The exercise group underwent a 4-month supervised training program followed by 4 months of home-based training. After the initial 4 months of training, patients in the exercise group showed a significant increase in peak VO(2) and reduction in MSNA, compared to the untrained group, but this was not maintained during 4 months of home-based training. In contrast, the decrease in FVR (56+/-3 vs. 46+/-4 vs. 40+/-2 U, p=0.008) and the improvement in QOL that were achieved during supervised training were maintained during home-based training. CONCLUSIONS: Home-based training following supervised training is a safe strategy to maintain improvements in QoL and reduction in FVR in chronic heart failure patients, but is an inadequate strategy to maintain fitness as estimated by peak VO(2) or reduction in neurohumoral activation. PMID- 16714147 TI - Primary care respiratory journal is awarded full Medline/Index Medicus listing. PMID- 16714148 TI - Epilepsy, intellectual disability, and living environment: a critical review. AB - Epilepsy occurs at a higher incidence and is more prevalent in people with an intellectual disability than in the general population. Nonetheless, we have insufficient knowledge of the extra needs of people with epilepsy and intellectual disability, of their families, and of the living environment. The lack of information about specific needs of the living environment may, in particular, be important. A Medline search revealed that scant attention has been paid to the specific needs for patients with epilepsy and intellectual disability, and only a few studies have focused specifically on this topic. The majority of studies have been focused on medical treatment issues and the organization and availability of health and social services. There is an indication that people with epilepsy and intellectual disability lack skills training appropriate to their intellectual potential. And although several reviews have emphasized the need for information on living environment and quality of life, we did not find such studies in our search. PMID- 16714149 TI - Electric field mediated DNA motion model. AB - Understanding the motion and the governing equations of a molecule's path in tissue is an ultimate requirement for the repeatable, site specific delivery of molecules [Joseph D. Hickey. Modelling the Motion of Ions and Molecules in Electroporation and Electrophoresis Field Conditions. University of South Florida, College of Arts and Sciences, Department of Physics, Tampa, Florida, 2003., Joseph D. Hickey and Richard Gilbert. Modeling the electromobility of ions in a target tissue. DNA and Cell Biology, 22 (12) (2003) 823-828.]. This paper describes a computationally efficient mathematical model and simulation technique for the examination of DNA fragments in a 1% agarose gel. The speed of the individual DNA fragments through the agarose gel was described through two parts. The maximum velocity was calculated using the Coulombic force divided by Stoke's law and that value was retarded by an exponential rate equation. The simulation utilizes previously published techniques modified for this specific application [Joseph D. Hickey and Richard Gilbert. Fluid flow electrophoresis model. Bioelectrochemistry, 63 (2) (2004) 365-367., Joseph D. Hickey and Richard Gilbert. Modeling the electromobility of ions in a target tissue. DNA and Cell Biology, 22 (12) (2003) 823-828.]. Five representative DNA fragment sizes that span the resolution of a 1% agarose gel were chosen for this analysis. The speeds corresponding to these five DNA fragment sizes were converted into discrete values and used in a 50 step simulation. The resultant error comparing the simulation with experimental distance was 7.76%. Through a 1-D optimization procedure, this error was reduced to 3.02% for a 52 step simulation. PMID- 16714150 TI - Mitochondrial disorders in renal tumors. AB - As early as 1930, Warburg discovered that metabolic alterations were associated with carcinogenesis and that cancer cells fermented even in the presence of oxygen using glycolysis to fulfill their energy needs, though less efficiently than with respiration. The kidney requiring a very active energy production for its pumping functions has a high mitochondrial activity. Kidney tumors can exist either in relatively benign forms, as for example, in oncocytomas that are crowded with mitochondria or in very aggressive forms such as in clear cell renal carcinomas that exhibit strongly down-regulated mitochondrial activities. These carcinomas can produce metastases that are resistant to anti-mitotic drugs and current treatments only delay the fatal issue. In this review, the mitochondrial alterations observed in various forms of renal tumors will be discussed with the aim of understanding how the knowledge of mitochondrial impairment mechanisms could be helpful to develop new anti-cancer strategies. PMID- 16714151 TI - The effect of incomplete acetylene washout on cardiac output measurement using open circuit acetylene uptake. AB - The open circuit acetylene uptake method is a useful non-invasive means of measuring cardiac output. However, because of accumulation of inhaled acetylene in tissues, the cardiac output uptake is underestimated, if residual acetylene is not allowed to wash out completely in between measurements. We determined the effect of applying a correction factor that estimates mixed venous acetylene concentration from endtidal values to the calculation of cardiac output. This accounts for mixed venous acetylene present during measurements made before complete washout. Six healthy subjects performed steady-state exercise at approximately 30% and 60% of V(O2 max). Cardiac output measurements were made at each exercise intensity using the open circuit acetylene uptake method (inspired [acetylene] approximately 1%), with the first and last measurements having no detectible levels of acetylene in expired gas (reference measurement). Data were also obtained with immediate pre-measurement endtidal concentrations ranging from 3% to 15% of the inspired [acetylene], in random order in between. Oxygen consumption, carbon dioxide production and heart rate did not change significantly during testing at each exercise intensity. Reference cardiac output also did not change significantly and averaged 11.1+/-0.8 L/min at 30% of V(O2 max) and 16.5+/-2.0 L/min at 60% of V(O2 max). Uncorrected cardiac output measurements progressively underestimated cardiac output by 15% at the 3% of inspired endtidal [acetylene] and by over 50% at 15% [acetylene] (p<0.0001). However, when corrected for residual endtidal [acetylene], cardiac outputs were not significantly different from the reference measurements. The results of this study suggest that by accounting for residual endtidal acetylene in mixed venous blood, cardiac output can be accurately measured even when washout of acetylene is incomplete, allowing measurements as often as every 10-15 s. PMID- 16714152 TI - Determination of 5-chloro-7-iodo-8-quinolinol (clioquinol) in plasma and tissues of hamsters by high-performance liquid chromatography and electrochemical detection. AB - This paper describes a method of determining clioquinol levels in hamster plasma and tissue by means of HPLC and electrochemical detection. Clioquinol was separated on a Nucleosil C18 300 mm x 3.9 mm i.d. 7 microm column at 1 ml/min using a phosphate/citrate buffer 0.1M (400 ml) with 600 ml of a methanol:acetonitrile (1:1, v/v) mobile phase. The retention times of clioquinol and the IS were, respectively, 11.6 and 8.1 min; the quantitation limit (CV>8%) was 5 ng/ml in plasma and 10 ng/ml in tissues. The intra- and inter-assay accuracies of the method were more than 95%, with coefficients of variation between 3.0 and 7.7%, and plasma and tissue recovery rates of 72-77%. There was a linear response to clioquinol 5-2000 ng/ml in plasma, and 10-1000 ng/g in tissues. The method is highly sensitive and selective, makes it possible to study the pharmacokinetics of plasma clioquinol after oral administration and the distribution of clioquinol in tissues, and could be used to monitor plasma clioquinol levels in humans. PMID- 16714153 TI - Monolithic organic polymeric columns for capillary liquid chromatography and electrochromatography. AB - This review briefly summarizes the present state of the preparation and use of capillary monolithic columns for liquid chromatography (LC) and electrochromatography (EC). Most important approaches to the preparation of monolithic stationary phases based on organic polymers are outlined and the properties of the monoliths obtained are compared with those of classical particulate phases. A few selected applications of monolithic columns are shown to demonstrate the most important advantages of monolithic capillary columns. It is concluded that both the monolithic and particulate capillary columns are important and that judicious choice of the type suitable for a particular application requires careful consideration of the purpose of the separation and the properties of the solutes to be separated. Monolithic columns are substantially younger than packed ones and thus will require further theoretical and experimental study to further improve their preparation and to enable reliable prediction of their properties and applicability; nevertheless, they are very promising for the future. PMID- 16714154 TI - Optimization of a capillary zone electrophoresis method by using a central composite factorial design for the determination of codeine and paracetamol in pharmaceuticals. AB - Capillary zone electrophoresis was optimized to quantitatively determine codeine and paracetamol via central composite factorial design. Critical parameters (concentration, buffer, pH, voltage) assessed effects on resolution, analysis time and efficiencies. Optimum separation conditions were achieved using phosphate buffer 20 mM (pH 6.8) and voltage (15 kV). The optimized procedure easily determined codeine and paracetamol with separation in less than 3 min. Calibration curves (R > 0.999) were prepared, with LODs of 13.5 and 340 ng mL(-1) for codeine and paracetamol, respectively, and a good R.S.D.% (<3%). This method was applied to determine codeine and paracetamol in pharmaceutical formulations; recoveries coincided with stated contents. PMID- 16714155 TI - Sensitive, rapid and validated gas chromatography/negative ion chemical ionization-mass spectrometry assay including derivatisation with a novel chiral agent for the enantioselective quantification of amphetamine-type stimulants in hair. AB - A novel chiral derivatisation agent, (2S,4R)-N-heptafluorobutyryl-4 heptafluorobutoyloxy-prolyl chloride, was used for the indirect resolution of amphetamine (AM), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA), 3,4 methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyethylamphetamine (MDEA) enantiomers using gas chromatography coupled to mass spectrometry operating in the negative-ion chemical ionization mode (GC/MS-NICI). This new chiral derivatisation reagent was readily obtained in optically pure form after a simple two-step synthesis. Optimal derivatisation was accomplished in 15 min at room temperature in a carbonate buffer and the resulting diastereoisomers were base line separated by GC in 12 min only. No racemization was observed during the derivatisation. The method was applied and fully validated for the enantiomeric quantification of amphetamines and methylenedioxylated amphetamines in hair. The analyses of 24 hair specimens from suspected ATS abusers showed that 24 cases were positive for MA and/or AM enantiomers and that in most cases the concentrations of (S)-MA and (S)-AM exceeded those of the corresponding (R) enantiomers. One hair specimen was tested positive for both enantiomers of MDMA and MDA. PMID- 16714156 TI - Simultaneous determination of cortisol and prednisolone in body fluids by using HPLC-DAD coupled with second-order calibration based on alternating trilinear decomposition. AB - A novel method for simultaneous determination of cortisol and prednisolone in body fluids has been developed in this paper. Three-way data recorded by high performance liquid chromatography with a diode array detector (HPLC-DAD) have been analyzed by second-order calibration based on the alternating trilinear decomposition (ATLD) algorithm. The chemometric methodology selected exploits the second-order advantage of the three-way data arrays, which allows one to obtain concentrations of individual calibrated analytes even in the presence of interferences not present in the calibration samples (e.g. background in urine or plasma). It was applied to simultaneous determination of cortisol and prednisolone in both plasma and urine samples. Though the chromatographic and spectral peaks of the analytes were heavily overlapped and interferents coeluted with the compounds studied, good recoveries of the analytes could be obtained with HPLC-DAD coupled with second-order calibration based on ATLD. Sample preparation was based on solvent extraction (SE), and quantification can be carried out with simple mobile phase. The time required for the quantification process is shorter than other methods. PMID- 16714157 TI - Novel adsorptive polyamine coating for enhanced capillary electrophoresis of basic proteins and peptides. AB - In capillary electrophoresis (CE), the anionic and hydrophobic nature of the fused-silica capillary surface has long been known to present a problem in protein and peptide analysis. The use of capillary surface coating is one of the approaches to avoid the analyte-wall interactions. In this study, a new polymer, poly-LA 313, has been synthesized, physico-chemical characterized, and applied as polyamine coating for CE separations. The coating process is highly reproducible and provides fast separations of peptides and proteins in a few minutes and with high efficiency. The physically adsorbed polymer gives rise to a durable coating in the range of pH 2-10, in the presence of organic modifiers (acetonitrile and methanol) and with complex biological samples. The efficiency of the new cationic polymer was also tested performing protein and peptide separations with capillary electrophoresis-electrospray ionization-mass spectrometry (CE-ESI-MS). PMID- 16714158 TI - Use of aldosterone antagonists in resistant hypertension. AB - Resistant hypertension is defined as an elevated blood pressure in spite of treatment with 3 different antihypertensive agents. The prevalence of resistant hypertension is unknown, but recent cross-sectional analyses and hypertension outcome studies suggest it is a common clinical problem and will become even more so with an aging and increasingly heavy population. Secondary causes of hypertension are common in patients with resistant hypertension, in particular, obstructive sleep apnea and hyperaldosteronism. Treatment of resistant hypertension is predicated upon identification and reversal of secondary causes of hypertension, as possible, and effective use of multidrug regimens. Recent clinical studies indicate that aldosterone antagonists, spironolactone and amiloride, provide significant additional blood pressure reduction when added to treatment regimens of patients with resistant hypertension. Both agents are generally well tolerated. Hyperkalemia is an uncommon complication of aldosterone antagonists, but it can occur; therefore, biochemical monitoring is necessary, particularly in high-risk patients. PMID- 16714159 TI - Systolic hypertension in older persons: how aggressive should treatment be? AB - Systolic hypertension (SH) is a major public health concern predominantly affecting older persons. A key message of the Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC 7) is that SH is a much more important cardiovascular disease risk factor than diastolic hypertension, particularly in older persons. Consequently, aggressive control of elevations of systolic blood pressure (SBP) is recommended. Despite increasing attention, SH is on the rise: isolated elevations of SBP in a national sampling of Veteran's Administration patients have increased from 57% in 1990 to 1995 to 76% of patients in 1999. This article considers several clinically pertinent issues, including the evidence for treating older patients with elevations in SBP, treating SH in the "oldest old" (those aged >85 years), and how aggressively these patients should be treated. In addition, issues regarding clinical decision making in older patients with SH are discussed. PMID- 16714161 TI - Fixed-drug combinations as first-line treatment for hypertension. AB - As combinations of drugs from different classes that have synergistic or additive effect and properties to cancel out each others' untoward hemodynamic and metabolic effects become more and more widely used, their use as first-line therapy for the treatment of newly diagnosed hypertensive patients is growing in popularity as well. The possibility to begin therapy with a fixed 2-drug combination may be preferable to starting with monotherapy followed by upward titration and addition of other agents. More and more combinations are coming out on the market and proving their effectiveness in randomized controlled trials and in large multicenter studies. One suggestion is the "polypill," a fixed combination of multiple agents that address various components of the metabolic syndrome and coexisting common risk factors in both high-risk patients with conditions requiring polypharmacy, and in healthy asymptomatic individuals. PMID- 16714160 TI - Baroreflex function in hypertension: consequences for antihypertensive therapy. AB - Studies performed in experimental animal models of hypertension as well as in human hypertension have unequivocally shown that reflex cardiovascular control undergoes profound changes with high blood pressure and participates throughout direct and indirect mechanisms at the development and progression of hypertension related target organ damage. This explains why investigation of the effects of antihypertensive drugs on neural cardiovascular control has a significant impact on hypertension treatment. This review will examine the main features of cardiovascular reflex control in hypertension and the effects of the different classes of antihypertensive drugs in uncomplicated and complicated hypertension. PMID- 16714162 TI - Ventricular arrhythmias and sudden cardiac death. PMID- 16714163 TI - South Africa needs an HIV truth and reconciliation commission. PMID- 16714164 TI - All that glisters is not gold. PMID- 16714165 TI - Independence at the US Food and Drug Administration. PMID- 16714166 TI - Clinical trial registration: transparency is the watchword. PMID- 16714167 TI - Trial registers: protecting patients, advancing trust. PMID- 16714168 TI - Trial summaries on results databases and journal publication. PMID- 16714169 TI - Avian influenza vaccines: what's all the flap? PMID- 16714170 TI - Dipyridamole with aspirin for secondary stroke prevention. PMID- 16714171 TI - Prediction versus prevention of sudden cardiac death. PMID- 16714172 TI - Modern physics for the modern physician. PMID- 16714173 TI - New science inspires FDA commissioner Andrew von Eschenbach. PMID- 16714174 TI - Statins and sepsis. PMID- 16714176 TI - Statins and sepsis. PMID- 16714177 TI - Effects of radiotherapy and surgery for early breast cancer. PMID- 16714178 TI - Effects of radiotherapy and surgery for early breast cancer. PMID- 16714179 TI - Effects of radiotherapy and surgery for early breast cancer. PMID- 16714180 TI - Effects of radiotherapy and surgery for early breast cancer. PMID- 16714182 TI - Spinning stars: ASTEROID's impact on atherosclerosis. PMID- 16714183 TI - Time for radical revision at the UK General Medical Council. PMID- 16714185 TI - Fruit and vegetable consumption and stroke. PMID- 16714186 TI - Safety and immunogenicity of an inactivated split-virion influenza A/Vietnam/1194/2004 (H5N1) vaccine: phase I randomised trial. AB - BACKGROUND: Pathogenic avian influenza A virus H5N1 has caused outbreaks in poultry and migratory birds in Asia, Africa, and Europe, and caused disease and death in people. Although person-to-person spread of current H5N1 strains is unlikely, the virus is a potential source of a future influenza pandemic. Our aim was to assess the safety and immunogenicity of a vaccine against the H5N1 strain. METHODS: We did a randomised, open-label, non-controlled phase I trial in 300 volunteers aged 18-40 years and assigned one of six inactivated split influenza A/Vietnam/1194/2004 (H5N1) influenza vaccine formulations, comprising 7.5 microg (with adjuvant n=50, without adjuvant n=49), 15 microg (n=50, n=50), or 30 microg (n=51, n=50) of haemagglutinin with or without aluminium hydroxide adjuvant. Individuals received two vaccinations (on days 0 and 21) and provided blood samples (on days 0, 21, and 42) for analysis by haemagglutination inhibition and microneutralisation. We recorded all adverse events. Analyses were descriptive. FINDINGS: All formulations were well tolerated, with no serious adverse events, few severe reactions, and no oral temperatures of more than 38 degrees C. All formulations induced an immune response, and responses were detectable in some individuals after only one dose. The adjuvanted 30 microg formulation induced the greatest response (67% haemagglutinin-inhibition seroconversion rate after two vaccinations). Adjuvant did not improve the response to the lower doses. Two vaccinations of non-adjuvanted 7.5 microg, adjuvanted 15 microg, or non adjuvanted 15 microg seroconverted more than 40% of participants (haemagglutinin inhibition test only). Haemagglutinin inhibition and neutralising results were comparable. INTERPRETATION: A two-dose regimen with an adjuvanted 30 microg inactivated H5N1 vaccine was safe and showed an immune response consistent with European regulatory requirements for licensure of seasonal influenza vaccine. We noted encouraging responses with lower doses of antigen that need further study to ascertain their relevance for the choice of the final pandemic vaccine. PMID- 16714187 TI - Aspirin plus dipyridamole versus aspirin alone after cerebral ischaemia of arterial origin (ESPRIT): randomised controlled trial. AB - BACKGROUND: Results of trials of aspirin and dipyridamole combined versus aspirin alone for the secondary prevention of vascular events after ischaemic stroke of presumed arterial origin are inconsistent. Our aim was to resolve this uncertainty. METHODS: We did a randomised controlled trial in which we assigned patients to aspirin (30-325 mg daily) with (n=1363) or without (n=1376) dipyridamole (200 mg twice daily) within 6 months of a transient ischaemic attack or minor stroke of presumed arterial origin. Our primary outcome event was the composite of death from all vascular causes, non-fatal stroke, non-fatal myocardial infarction, or major bleeding complication, whichever happened first. Treatment was open, but auditing of outcome events was blinded. Primary analysis was by intention to treat. This study is registered as an International Standard Randomised Controlled Trial (number ISRCTN73824458) and with (NCT00161070). FINDINGS: Mean follow-up was 3.5 years (SD 2.0). Median aspirin dose was 75 mg in both treatment groups (range 30-325); extended-release dipyridamole was used by 83% (n=1131) of patients on the combination regimen. Primary outcome events arose in 173 (13%) patients on aspirin and dipyridamole and in 216 (16%) on aspirin alone (hazard ratio 0.80, 95% CI 0.66-0.98; absolute risk reduction 1.0% per year, 95% CI 0.1-1.8). Addition of the ESPRIT data to the meta-analysis of previous trials resulted in an overall risk ratio for the composite of vascular death, stroke, or myocardial infarction of 0.82 (95% CI 0.74-0.91). Patients on aspirin and dipyridamole discontinued trial medication more often than those on aspirin alone (470 vs 184), mainly because of headache. INTERPRETATION: The ESPRIT results, combined with the results of previous trials, provide sufficient evidence to prefer the combination regimen of aspirin plus dipyridamole over aspirin alone as antithrombotic therapy after cerebral ischaemia of arterial origin. PMID- 16714188 TI - Deceleration capacity of heart rate as a predictor of mortality after myocardial infarction: cohort study. AB - BACKGROUND: Decreased vagal activity after myocardial infarction results in reduced heart-rate variability and increased risk of death. To distinguish between vagal and sympathetic factors that affect heart-rate variability, we used a signal-processing algorithm to separately characterise deceleration and acceleration of heart rate. We postulated that diminished deceleration-related modulation of heart rate is an important prognostic marker. Our prospective hypotheses were that deceleration capacity is a better predictor of risk than left-ventricular ejection fraction (LVEF) and standard deviation of normal-to normal intervals (SDNN). METHODS: We quantified heart rate deceleration capacity by assessing 24-h Holter recordings from a post-infarction cohort in Munich (n=1455). We blindly validated the prognostic power of deceleration capacity in post-infarction populations in London, UK (n=656), and Oulu, Finland (n=600). We tested our hypotheses by assessment of the area under the receiver-operator characteristics curve (AUC). FINDINGS: During a median follow-up of 24 months, 70 people died in the Munich cohort and 66 in the London cohort. The Oulu cohort was followed-up for 38 months and 77 people died. In the London cohort, mean AUC of deceleration capacity was 0.80 (SD 0.03) compared with 0.67 (0.04) for LVEF and 0.69 (0.04) for SDNN. In the Oulu cohort, mean AUC of deceleration capacity was 0.74 (0.03) compared with 0.60 (0.04) for LVEF and 0.64 (0.03) for SDNN (p<0.0001 for all comparisons). Stratification by dichotomised deceleration capacity was especially powerful in patients with preserved LVEF (p<0.0001 in all cohorts). INTERPRETATION: Impaired heart rate deceleration capacity is a powerful predictor of mortality after myocardial infarction and is more accurate than LVEF and the conventional measures of heart-rate variability. PMID- 16714189 TI - Tuberculous meningitis. PMID- 16714190 TI - Systemic sclerosis: hypothesis-driven treatment strategies. AB - We review data from controlled trials and randomised controlled trials to examine the hypothesis for the pathogenesis of systemic sclerosis. Strategies used to treat the vascular complications in systemic sclerosis have so far shown the biggest successes, especially in the management of renal crisis and pulmonary arterial hypertension. Because these drugs have improved function and quality of life and have increased survival rates, they can truly be classified as disease modifying compounds. Immunosuppressive therapy with cyclophosphamide in particular has also shown evidence of efficacy, and randomised controlled trials of autologous stem-cell transplantation are underway. So far, strategies to reduce or control fibrosis directly (bosentan, interferon gamma, and relaxin) have been disappointing but new strategies against fibrosis based on advanced understanding of the molecular biology of systemic sclerosis hold promise. Treatments against several cardinal features of the disorder simultaneously have not yet been examined but are being considered for future trials. PMID- 16714191 TI - Astroparticle physics and cosmology. AB - Astroparticle physics is an interdisciplinary field that explores the connections between the physics of elementary particles and the large-scale properties of the universe. Particle physicists have developed a standard model to describe the properties of matter in the quantum world. This model explains the bewildering array of particles in terms of constructs made from two or three quarks. Quarks, leptons, and three of the fundamental forces of physics are the main components of this standard model. Cosmologists have also developed a standard model to describe the bulk properties of the universe. In this new framework, ordinary matter, such as stars and galaxies, makes up only around 4% of the material universe. The bulk of the universe is dark matter (roughly 23%) and dark energy (about 73%). This dark energy drives an acceleration that means that the expanding universe will grow ever larger. String theory, in which the universe has several invisible dimensions, might offer an opportunity to unite the quantum description of the particle world with the gravitational properties of the large scale universe. PMID- 16714192 TI - Reversing the decline of academic medicine in Europe. PMID- 16714194 TI - Tropical dermatology. PMID- 16714193 TI - Necrobacillosis without Lemierre's syndrome. PMID- 16714195 TI - The history of smallpox. AB - The history of smallpox, from its probable origin 3000 to 6000 years ago to its worldwide eradication in 1978, is a fascinating story. A feared infection, even today, smallpox has caused the deaths of millions of people and disfigured many. This article describes the possible origin of the infection and traces its progress throughout the world. The introduction of inoculation with material from smallpox lesions and vaccination using cowpox virus is described. The World Health Organization's successful smallpox eradication program in the 1960s and 1970s is described. Personal experience of this program in Bangladesh in 1973 is outlined, as is the laboratory escape of the major smallpox virus in Birmingham in 1978. The possible use of the smallpox virus as a biological weapon is discussed. PMID- 16714196 TI - Allergic skin reactions in the tropics. AB - Cutaneous allergic reactions are relatively common in tropical countries, with some caused by the tropical environment and others by changes in the traditional living conditions combined with poverty. HIV infection is a major contributory factor. A large number of reactions are eczematous. Atopic dermatitis is on the increase together with contact allergic reactions. Available products are often of inferior quality and contain irritants and allergens that may be forbidden elsewhere in the world. The extensive and uncontrolled use of drugs and indigenous medications together with an increase in HIV prevalence leads to drug eruptions such as the Stevens-Johnson syndrome and toxic epidermal necrolysis. Poverty plays an important role in the prevalence of insect bites and resulting papular urticaria and possibly in acropustulosis infantum. Many unexplained environmentally induced blistering diseases occur, ranging from phytophotodermatitis to fogo selvagem and from pemphigus foliaceus to chronic bullous dermatosis of childhood. This article describes the allergic conditions that occur in the tropics. Special attention is given to those cases of eczema and eruptions that are specific to tropical countries. Available treatments are also discussed. PMID- 16714197 TI - Itch and scratch: scabies and pediculosis. AB - Scabies and lice infestations are almost an everyday diagnosis in the dermatology clinics. Along with the unbearable itch, resistance to medication, secondary infection, and the high risk for spreading the parasite to their close contacts, the patients have also to battle with many myths, prejudice, and shames connected to those infestations. PMID- 16714198 TI - Onchocerciasis--river blindness. AB - Onchocerciasis results from infestation by the nematode Onchocerca volvulus and is characterized by troublesome itching, skin lesions, and eye manifestations. Although partially controlled by international mass prevention programs, onchocerciasis remains a major health hazard and is endemic in Africa, Arabia, and the Americas. Onchocerciasis is spread by bites from infested black flies, which transmit larvae that subsequently develop into adult filariae. Skin symptoms are commonly nonspecific and include severe pruritus, acute and chronic dermatitis, vitiligo-like hypopigmentation, and atrophy. Onchocercal ocular disease covers a large spectrum of manifestations, which in severe cases, may lead to blindness. Diagnosis is usually made by direct visualization of the larvae emerging from superficial skin biopsies, "skin snips." In some cases, the microfilariae can also be directly observed at the slit lamp when migrating into the anterior chamber of the eye. Ivermectin is, at present, the drug of choice for skin and ocular manifestations. Recent research using a chemotherapeutic approach that targets filarial Wolbachia symbionts in the treatment and control of onchocerciasis, however, suggests that 100 mg/d of doxycycline for 6 weeks might be effective in reducing the filarial load and preventing ocular symptoms. PMID- 16714200 TI - Tropical rickettsioses. AB - In recent years, a bewildering array of emerging rickettsial pathogens have been described throughout the world, including in the tropics. Here we present an updated overview of scrub typhus, murine typhus, and epidemic typhus. We also present an update on the emerging spotted fever group rickettsioses described in the tropics through 2005, focusing on epidemiologic and clinical data and management. PMID- 16714199 TI - Endemic treponematosis: review and update. AB - Despite major efforts to eradicate these disorders, yaws, bejel, and pinta (endemic treponematosis) remain serious health issues in many regions of the world. Aside from prominent skin manifestations, these diseases may also lead to significant osseous, neurologic, and ophthalmologic complications. Although progress has been made in differentiating the causative species in a research setting, a simple, specific, and sensitive diagnostic test remains elusive. Parenteral penicillin, in appropriate dosage, is the treatment of choice; alternative antibiotics such as tetracycline and erythromycin may also be effective. PMID- 16714201 TI - Fungal infections. AB - Tropical mycoses vary from the commonplace to the rare and exotic but they are seen regularly even as imported conditions and it is important to consider the diagnosis, where possible, in individuals who have visited remote areas. Patients may present many years after they have left an endemic area with subcutaneous or endemic systemic mycoses. In establishing a diagnosis, it is always important to take an accurate travel history so that the movements of the individual can be correlated with the potential risk for exposure to infection. PMID- 16714202 TI - Loxoscelism. AB - Loxoscelism (bites by spiders of the genus Loxosceles) is the only proven arachnological cause of dermonecrosis. Although Loxosceles spiders can be found worldwide, their distribution is heavily concentrated in the Western Hemisphere, particularly the tropical urban regions of South America. Although Loxosceles bites are usually mild, they may ulcerate or cause more severe, systemic reactions. These injuries mostly are due to sphingomyelinase D in the spider venom. There is no proven effective therapy for Loxosceles bites, although many therapies are reported in the literature. PMID- 16714203 TI - Personal observation of skin disorders in malnutrition. AB - This is a description of some unknown skin disorders found by a physician inmate in a concentration camp, 1958 to 1962. After prolonged semistarvation and ultraheavy physical labor, skin lesions developed among the inmates including cutaneous pigmentation overlying bony prominence, buccal membrane pigmentation, palmoplantar keratoderma with fissures, palmar crease clefts, nail layering, intra-nail hemorrhage, and so on. These lesions responded dramatically to nutrition therapy, including dietary improvement, yeast administration, or thiamin injection. Thiamin deficiency was confirmed to be one of major etiologic factors, whereas the deficiency of niacin or riboflavin also played a part. In the pediatric case with palmar crease clefts, both thiamin and niacin were dramatically effective. No laboratory data could be provided. PMID- 16714204 TI - Tachyphylaxis to topical corticosteroids: the more you use them, the less they work? PMID- 16714206 TI - Editorial. PMID- 16714207 TI - The effect of overexpression of endothelial nitric oxide synthase on eosinophilic lung inflammation in a murine model. AB - The effects of nitric oxide (NO) on allergic inflammation are controversial. In particular, the role of endothelial nitric oxide synthase (eNOS) in asthma remains uncertain. In the present study, we examined the effects of overexpression of eNOS on allergic inflammation using eNOS transgenic (eNOS-Tg) mice, in which eNOS protein is overexpressed in the vascular endothelium and airway epithelium. We found that eNOS-Tg mice showed a reduction of the asthmatic response to allergen challenge. Eosinophilic accumulation in the airspaces, eosinophilic activity, and bronchial responsiveness to acetylcholine were significantly attenuated in eNOS-Tg mice, as compared with wild-type mice following ovalbumin sensitization/challenge, even though the levels of circulating eosinophils were comparable in the wild-type and eNOS-Tg mice. The concentrations of eotaxin in the bronchoalveolar lavage fluid were significantly less in eNOS-Tg mice than in the wild-type mice. In addition, immunohistochemical analysis showed that the expressions of both intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 on the pulmonary endothelium of eNOS-Tg mice was decreased compared with the controls. These results suggest that chronic eNOS overexpression contributes to the suppression of allergic inflammation by reducing the production of eotaxin in the airspaces and/or the expression of adhesion molecules in the vascular endothelium. PMID- 16714208 TI - Propolis extracts exhibit an immunoregulatory activity in an OVA-sensitized airway inflammatory animal model. AB - Propolis, which has been used widely in folk medicine, has been shown to exhibit various biological activities but its immunoregulatory and anti-inflammatory activities in intact animals have not been well studied. We investigated these activities of propolis using an ovalbumin-induced asthma animal model. Mice were immunized and sensitized by exposure to ovalbumin (OVA) antigen and administered with low- (65 mg/kg body weight) and high-dose (325 mg/kg body weight) propolis water extracts by tube feeding. The serum OVA-specific IgE titer and cytokine profiles in cultured splenocytes and bronchoalveolar lavage fluids (BALF) were analyzed. The number of eosinophils in BALF was counted. Here we demonstrate that propolis extracts can suppress the serum levels of OVA-specific IgE and IgG(1), and airway hyperresponsiveness (AHR) in OVA-sensitized mice. There are no significant differences in the concentration of eotaxin or the number of eosinophils in BALF among the four groups. However, the higher dose of propolis extracts decreases the level of IL-5 in BALF. The splenocytes from mice administered with propolis extracts (low- and high-dose groups) exhibit a strong inhibition of IL-10 secretion and up-regulation of IFN-gamma secretion in splenocytes stimulated with concanavalin A (ConA). In addition, cytokine (IFN gamma, IL-6, and IL-10) secretion in OVA-stimulated splenocytes from the propolis groups was significantly lower than that in the control group. These results suggest that propolis extracts may be a potential novel therapeutic agent for asthma. PMID- 16714209 TI - Modulation of neutrophil apoptosis by beta-amyloid proteins. AB - This study examined the effect of amyloid beta peptide (Abeta) and the secretase inhibitors of amyloid precursor proteins (APP) on the spontaneous apoptosis of neutrophils. Abeta(1-40) decreased the apoptotic rate of neutrophils. The delayed apoptosis by Abeta was not blocked by pertussis toxin and N-formyl peptide receptor-like 1 antagonistic peptide, WRWWWW. The inhibitors of phoshoinositide 3 kinase (LY294002), phospholipase C (U73122), or Ca++-dependent protein kinase C (Go6976) abrogated the anti-apoptotic effect of Abeta on neutrophils. Moreover, the Abeta-induced delay of apoptosis was inhibited by a calcium chelator, BAPTA/AM. The amount of the APP protein was reduced in the cultured neutrophils and the APP level in the Abeta or pancaspase-treated neutrophils was lower than that in the cultured neutrophils. However, the reduction in APP level was recovered after treating them with the secretase inhibitors or anti-Fas antibody. The exogenous addition of cell permeable beta- and gamma-secretase inhibitors resulted in an increase in the rate of the apoptosis. The regulation of neutrophil apoptosis by the addition of Abeta and secretase inhibitors occurred via the caspase -8, -9, -3, and mitochondrial-dependent pathways. This suggests that the intracellular beta-amyloid proteins play a role as regulating factor of neutrophil survival and that Abeta-induced delay of apoptosis is mediated by other receptors rather than a seven-transmembrane G protein-coupled receptor(s). PMID- 16714210 TI - Immunomodulatory properties of Cumaside. AB - The medical lead, so-called Cumaside, was created on the basis of triterpene oligoglycosides from the Far-Eastern edible sea cucumber (holothurian) Cucumaria japonica and its immunomodulatory properties were studied. The haemolytic activity of Cumaside was significantly reduced in comparison with original glycosides due to the glycoside-cholesterol complex formation. The influence of Cumaside on mouse macrophages in low doses was accompanied by more then two-fold stimulation of lysosomal activity. This preparation was found to increase significantly the animal resistance against bacterial infections elicited by various pathogens. It stimulated phagocytosis, ROS formation, IL6 and TNF-alpha production in lymphocytes, increased the number of antibody producing cells and amplified the expression of several cell surface molecules (CD3, CD4, CD8) preliminary cultured with hydrocortisone. At the same time the preparation did not affect the delayed-type hypersensitivity, proliferative activity of lymphocytes, cytotoxic activity of NK-cells and cytokine IFNgamma and IL12p70 release. The mechanism of Cumaside action is discussed. PMID- 16714211 TI - Interaction among vitamin D(3) analogue KH 1060, TNF-alpha, and vitamin D receptor protein in peripheral blood mononuclear cells of inflammatory bowel disease patients. AB - BACKGROUND: The active form of vitamin D, 1,25(OH)(2)D(3), exerts important effects on proliferation and differentiation of many cell types, and immunoregulatory activities in particular on T cell-mediated immunity. AIM: The aim of this study was to investigate whether KH 1060, a vitamin D analogue, could decrease tumor necrosis factor-alpha (TNF-alpha) levels in patients with inflammatory bowel disease (IBD). METHODS: PBMC proliferation was determined by [(3)H]thymidine incorporation. TNF-alpha levels were measured by ELISA kit; VDR, Bcl-2 and Bax protein levels with Western blot analysis. RESULTS: KH 1060 inhibited PBMC proliferation and decreased TNF-alpha levels in IBD patients and this effect was synergistic with anti-TNF-alpha. VDR protein levels were significantly increased by PBMC treatment with KH 1060 or anti-TNF-alpha or their combination in ulcerative colitis (UC) patients, and decreased in Crohn's disease (CD) patients, treating the cells with KH 1060. In UC patients an increase in Bcl 2 and Bax levels was observed incubating, PBMC with KH 1060 or anti-TNF-alpha or their combination. In CD patients a slight decrease in Bcl-2 levels was registered when anti-TNF alone or in association with KH 1060 was used. Bax protein levels were slightly increased in the presence of KH 1060 alone or in combination with anti-TNF. CONCLUSION: This study shows that KH 1060 acts as an immunomodulator on PBMC, acting as TNF-alpha inhibitor. This finding provides strong evidence that vitamin D status could be an important regulator of immunity IBD. PMID- 16714212 TI - Oral administration of exopolysaccharide from Aphanothece halophytica (Chroococcales) significantly inhibits influenza virus (H1N1)-induced pneumonia in mice. AB - The halophilous cyanobacterium Aphanothece halophytica releases large sums of single type sulfated exopolysaccharide in late logarithmic growth phase in culture. This polysaccharide contained sulfate up to 34.46% of the total moieties in the molecular. As a sulfated polysaccharide that can be biosynthesized in large quantities, however, its antiviral activity has not yet been reported. In this study, we examined effects of exopolysaccharide from A. halophytica Fremy (EPAH) on influenza virus A FM (H1N1) (FM1)-induced pneumonia and reduction in immunocompetence in mice. Previous and simultaneous treatment of EPAH at a dose of 60 mg/kg significantly inhibited pneumonia in FM1-infected mice by 30.4% and 26.7%, respectively. In post-treatment, EPAH displayed its most effective inhibition at a dose of 80 mg/kg with the inhibition rate at 18.69%. Simultaneous treatment of FM1-infected mice with EPAH showed effective improvement on reduction of lymphocyte number with its most effective dose at 60 mg/kg. FM1 infected mice simultaneously received EPAH at a dose of 40 mg/kg also acquired obvious enhancement on release of IL-2 on day 15, and those received EPAH at a dose of 60 mg/kg showed similar enhancement on day 10. Simultaneous treatment with EPAH indicated remarkable recovery or improvement of FM1-induced reduction of IL-1beta level and phagocytic capacity of RES. Simultaneous treatment with EPAH significantly resumed the cytolytic activity of natural killer cells in FM1 infected or CP treated mice at doses of 40 and 60 mg/kg. These results suggested that EPAH is an effective agent against FM1. The mechanisms of its action might be mediated, at least in part, by modulating the host immune system and the interaction positive charges in EPAH and negative charges FM1. PMID- 16714213 TI - Impact of interleukin-1 and interleukin-6 in murine primary schistosomiasis. AB - BACKGROUND: Immunization with schistosome antigens invariably elicits a plethora of cytokines and, hence, it is reasonable to assume that these cytokines influence host responses to challenge lung-stage larvae and, consequently, the adult worm burden, and may be responsible for the erratic data generally observed in protection studies against schistosome infection. METHODS: Schistosoma mansoni infected mice were administered with recombinant interleukin (IL)-1beta or IL-6 to evaluate the impact of cytokines in host responses to lung-stage schistosomula, and subsequent effects on adult worm parameters. Plasma lipid levels were assayed by colorimetric enzymatic tests and antibody responses by ELISA. Cytokine profile in peripheral blood mononuclear cells was evaluated by RT PCR. RESULTS: S. mansoni infection elicited, at the time of parasite residency in the lung, significant increase in free fatty acids (FA) and decrease in cholesterol plasma levels in C57BL/6 and CD1 mice, and stimulation of mRNA expression for cytokines of T helper type (Th) 2 in BALB/c, Th1 in C57BL/6, and Th1/Th2 in CD1 mice. However, no specific antibody production was evident in any mouse strain. In BALB/c mice, exogenous IL-1beta-related plasma free FA level significant increase, stimulation of expression of IL-1 and IL-12 mRNA, and considerable increase in percent of specific antibody-producing mice were associated with significant reduction in adult worm burden and egg load. In contrast, exogenous IL-1beta elicited decrease in free FA plasma levels, and down regulation of cytokines' mRNA expression in C57BL/6 and CD1 mice, changes associated with aggravation of the worm burden. Likewise, exogenous IL-6 failed to stimulate increase in plasma free FA levels or percent of antibody-producing mice except in BALB/c mice, effects that were protective for the host in BALB/c and for the parasite in C57BL/6 and CD1 mice. CONCLUSION: These findings were discussed in relation to the erratic data of protection experiments with schistosome subunit antigens in different mouse strains. PMID- 16714214 TI - C5a mediates secretion and activation of matrix metalloproteinase 9 from human eosinophils and neutrophils. AB - Matrix metalloproteinase 9 (MMP-9) is a crucial proteinase, utilized by both eosinophils and neutrophils, that mediates transmigration through extracellular basement membranes. We have found that neutralization of MMP-9 by a monoclonal antibody or a chemical inhibitor blocked C5a dependent chemotaxis of these granulocytes in vitro. The levels of MMP-9 secreted by the action of C5a from eosinophils were about 50-fold lower than those from neutrophils, consistent with results from confocal microscopy, where the density of MMP-9 containing granules was fewer within eosinophils than in neutrophils. Zymography indicated gelatin degrading activity of the molecular size of pro MMP-9 in supernatants from eosinophils and neutrophils stimulated by C5a, with no evidence of proteolytic activation. Instead MMP-9 activation appeared oxidative, since inhibition of NADPH oxidase and nitric oxide synthase by DPI or L-NIL abrogated C5a-mediated chemotaxis through basement membranes. In keeping with this mode of activation, C5a, known as an agent of superoxide generation, was also found to induce secretion of nitric oxide from human eosinophils and rat granulocytes and monocytes. In conclusion C5a is an important mediator that brings about secretion and oxidative activation of MMP-9, a requisite protease for transmigration, from both eosinophils and neutrophils. PMID- 16714215 TI - Effect of endothelins on human neutrophil activation by immune complexes. AB - Neutrophils are important effector cells of tissue injury in several pathological conditions, among them, immune complexes (IC)-induced inflammation and tissue injury. There is evidence that endothelins modulate IC-induced tissue injury in experimental models in vivo. In the present study we investigated the effect of endothelins on neutrophil activation by IC in vitro. To this purpose, pre-formed insoluble immune complexes were used to stimulate human neutrophils and production of leukotriene B(4) (LTB(4)) and hydrogen peroxyde (H(2)O(2)) were measured as indicative of phospholipase A(2) and oxidative burst activation and myeloperoxidase (MPO) release as indicative of cell degranulation. The effect of endothelins (ETs) in these events induced by IC was then examined. We found that IC stimulated all three events in human neutrophils. Addition of ET-1 but not ET 2 or ET-3 to the IC-stimulated neutrophils potentiated LTB(4) but not H(2)O(2) production. The endothelins added to resting neutrophils did not induce LTB(4) production but they were effective to stimulate H(2)O(2) production. The increased MPO activity induced by IC was not affected by endothelins nor did they stimulate the release of this enzyme in resting cells. These results show that endothelins are able to activate some neutrophil functions and to upregulate the IC-induced production of the pro-inflammatory molecule LTB(4). These data indicate that products of endothelial cells, such as endothelins, can be involved in the potentiation of neutrophil-dependent tissue injury. PMID- 16714216 TI - Inhibited cell death, NF-kappaB activity and increased IL-10 in TCR-triggered thymocytes of transgenic mice overexpressing the glucocorticoid-induced protein GILZ. AB - Glucocorticoids promote thymocyte apoptosis and modulate transcription of several genes including GILZ, which is strongly up-regulated in the thymus. We used transgenic mice overexpressing GILZ in the T-cell lineage to investigate TCR triggered functions of GILZ-overexpressing thymocytes. TCR-triggered apoptosis, but not glucocorticoid-induced apoptosis, was inhibited in transgenic mice compared to their controls. In vivo anti-CD3 administration did not reduce CD4(+)CD8(+) thymocyte number. Analysis of TCR-triggered molecular changes indicated that p65 NF-kappaB nuclear translocation and DNA binding activity was inhibited in transgenic mice, which might be linked with apoptosis inhibition. IL 10 release increased whereas release of IL-2, IFN-gamma, IL-13 and IL-4 remained unchanged. These results support the hypothesis that GILZ regulates, at least in part, T-cell development by influencing thymus function at cellular and molecular levels. PMID- 16714217 TI - Anti-inflammatory effect of thymoquinone in a mouse model of allergic lung inflammation. AB - Thymoquinone (TQ), the main active constituent of the volatile oil extracted from Nigella sativa's seeds, has been reported to have an anti-inflammatory and immune stimulatory effect on bronchial asthma and inflammation. However, little is known about the factors and mechanisms underlying these effects. In the present study, we examined the effect of TQ on airway inflammation in a mouse model of allergic asthma. Intraperitoneal injection of TQ before airway challenge of ovalbumin (OVA)-sensitized mice resulted in a marked decrease in lung eosinophilia and the elevated Th2 cytokines observed after airway challenge with OVA antigen; both in vivo, in the bronchoalveolar lavage (BAL) fluid and in vitro, following stimulation of lung cells with OVA. TQ also decreased the elevated serum levels of OVA-specific IgE and IgG1. Histological examination of lung tissue demonstrated that TQ significantly inhibited allergen-induced lung eosinophilic inflammation and mucus-producing goblet cells. While TQ showed a significant effect in inhibiting IL-4, IL-5 and IL-13 and some effect in inducing IFN-gamma production in the BAL fluid, it did show a slight effect on in vitro production of IL-4 by cultured lung cells stimulated with OVA antigen. These data suggest that TQ attenuates allergic airway inflammation by inhibiting Th2 cytokines and eosinophil infiltration into the airways; thus demonstrating its potential anti inflammatory role during the allergic response in the lung. PMID- 16714218 TI - Anergic lymphocytes generated by blocking CD28 and ICOS pathways in vitro prolong rat cardiac graft survival. AB - Regulatory cells may play a pivotal role in inducing and maintaining transplantation tolerance. We investigated the mechanism of anergic lymphocytes with regulatory cell potential generated in vitro by ICOS and CD 28 co stimulatory blockades as a source of cellular therapy for treating allograft rejection. Anergic lymphocytes were generated by a mixed lymphocyte reaction consisting of DA splenocytes as the stimulator and Lewis splenocytes as the responder in the presence of anti-ICOS mAb and rCTLA-4I g. Immunoregulatory effects of these lymphocytes were evaluated by secondary MLR and using other various stimulations. DA heart was transplanted into 7.5 Gy-irradiated Lewis rat after intravenous administration of these cells and/or Lewis spleen lymphocytes. We observed that these lymphocytes were not only anergic to alloantigen and polyclonal stimulations but also exhibited regulative activity to inhibit the alloreactive T-cell response. Our adoptive transfer studies revealed that irradiated recipients that received both anergic lymphocytes and naIve Lewis lymphocytes had significantly prolonged DA cardiac graft survival (mean 17.5 days) compared with a group that received Lewis lymphocytes alone (mean 10.8 days). Furthermore, some of the recipients accepted the graft indefinitely after receiving anergic lymphocytes alone (>100 days). These results demonstrated that anergic lymphocytes with regulatory activities can be generated through blocking co-stimulatory signals, CD 28 and ICOS, simultaneously in vitro, and may advance a new immunomodulatory strategy for preventing allorejection in organ transplantation. PMID- 16714219 TI - Anti-allergic properties of Cissampelos sympodialis and its isolated alkaloid warifteine. AB - Development of new agents capable of regulating eosinophilic inflammation can uncover novel therapeutic approaches for the treatment of allergic diseases, such as asthma. Here, we evaluated the anti-allergic properties of an extract of the Brazilian Menispermaceae Cissampelos sympodialis, focusing on its effects on allergic eosinophilia. By studying two models of allergic inflammation, an asthma model and the allergic pleurisy in actively sensitized Balb/c mice, we observed that the oral pre-treatment with C. sympodialis reduced pleural eosinophil influx triggered by allergen challenge in a dose-dependent manner. The mechanism involved in C. sympodialis inhibitory effect appeared to be independent of a direct effect on eosinophil locomotory machinery, but depend on a blockage of eotaxin production, a key eosinophil chemoattractant with important roles in allergic reactions. C. sympodialis was also able to affect eosinophil activation, as attested by its ability of inhibiting formation of new cytoplasmic lipid bodies and the secretion of cysteinyl leukotrienes. The alkaloid warifteine isolated from the C. sympodialis extract represents an active component responsible for the anti-eosinophilic effects of the extract, since warifteine was able to reproduce C. sympodialis inhibitory effects on allergic eosinophilia and cysteinyl leukotrienes production. Of interest, C. sympodialis and warifteine post-treatments also effectively inhibited eosinophilic reaction observed after allergic challenge. Therefore, C. sympodialis/warifteine may be a promising anti allergic therapy, inasmuch as it presents potent anti-eosinophil and anti leukotrienes activities. PMID- 16714220 TI - Regulation on maturation and function of dendritic cells by Astragalus mongholicus polysaccharides. AB - Astragalus mongholicus polysaccharides(ASP) isolated from one of the Chinese herbs-A. mongholicus which are known to have a variety of immunomodulatory activity. However, little is known about their immunomodulatory effects on murine bone marrow (BM)-derived dendritic cells (DC). DC are professional antigen presenting cells, which are pivotal for initiation of primary immune response. In this study, the regulatory effects of ASP on maturation and function of cultured murine BM-derived DC were investigated in vitro. ASP (10, 50, 100, 250 microg/ml) could increase the co-expression of CD-11c and MHC class II molecules on DC surface, and the 100 microg/ml is the optimal dose. The ability of unstimulated DC to uptake FITC-dextran was higher than that of ASP- or LPS-treated DC. We analyzed the concentration of IL-12 secreted by DC using ELISA. ASP-treated DC secreted a higher level of IL-12 than untreated DC. And ASP- or LPS-treated DC displayed a more mature morphology, with long protrusions, while untreated-DC displayed shorter protrusions than stimulated DC. PMID- 16714221 TI - Fraxetin inhibits the induction of anti-Fas IgM, tumor necrosis factor-alpha and interleukin-1beta-mediated apoptosis by Fas pathway inhibition in human osteoblastic cell line MG-63. AB - The survival of osteoblast cells is one of the determinants of the development of osteoporosis in patients with inflamed synovium, such as in rheumatoid arthritis (RA). By means of alkaline phosphatase (ALP) activity and osteocalcin ELISA assay, we have shown that fraxetin exhibits a significant induction of differentiation in the human osteoblast-like cell line MG-63. In addition, we also assessed whether fraxetin affects inflammatory cytokine-mediated apoptosis in osteoblast cells. TNF-alpha or IL-1beta enhance apoptotic DNA fragmentation in anti-Fas IgM-treated MG-63 cells by increasing Fas receptor expression. However, TNF-alpha or IL-1beta treatment alone does not induce apoptosis. Treatment of MG 63 cells with fraxetin not only inhibited anti-Fas IgM-induced apoptosis, but also blocked the synergetic effect of anti-Fas IgM with TNF-alpha or IL-1beta on cell death. The apoptotic inhibition of fraxetin is associated with inhibition of TNF-alpha and IL-1beta-mediated Fas expression and enhancement of FLIP expression, resulting in a decrease of caspase-8 and caspase-3 activation. These results indicate a potential use of fraxetin in preventing osteoporosis by inhibiting inflammatory cytokine-mediated apoptosis in osteoblast cells. PMID- 16714222 TI - Reverse of NK cytolysis resistance of type II cytokine predominant-human tumor cells. AB - Tumor cells suppress host immunity by secreting soluble factors, among which cytokines play important roles. Type I cytokine mRNA levels (IL-2 and IFNgamma) and type II cytokine mRNA levels (IL-4, IL-10 and IL-13) of twenty tumor cell lines and fresh tumor tissues from fifteen patients with colorectal cancer were examined by RT-PCR method. Upon analysis, tumor cells showed type II predominance and were resistant to NK cytolysis. In order to reverse the type II predominant status, type I cytokines (IL-2 or IFNgamma) and/or anti-type II cytokine antibody (IL-10 mAb) were used in an in vitro study. By this shifting treatment, type II cytokine mRNA levels of these tumor cells were decreased, moreover, the sensitivity to natural cytolysis of these cells was then significantly increased. Thus, tumor-induced type II cytokine dominance is possibly to be shifted into type I status, which benefits the anti-tumor natural immunity. PMID- 16714223 TI - Cellular and humoral systemic and mucosal immune responses stimulated in volunteers by an oral polybacterial immunomodulator "Dentavax". AB - The oral polybacterial immunomodulator Dentavax (D), composed of killed cells from Klebsiella pneumoniae, Streptococcus pyogenes, Staphylococcus aureus, Candida albicans and Lactobacillus acidophilus and their lysates was created for immunoprophylaxis and therapy of oral mucosa and parodont inflammations. The stimulating effect of the preparation was evaluated in twelve volunteers immunized for 10 consecutive days. On days 7, 14, 21, 28 and 49 after the last immunization peripheral blood (PB) lymphocyte subsets, T lymphocyte activation and PB phagocytic activity, were studied by flow cytometry. PB lymphocyte proliferative responses to PHA, rIL-2, LPS and D were evaluated radiometrically. The production of TNF-alpha in supernatants of in vitro stimulated lymphocytes and specific IgA, IgM and IgG antibodies in serum and saliva was determined by ELISA. Ultrastructural morphologic changes in T and B lymphocyte populations were also investigated. Although no significant changes in the levels of basic lymphocyte subsets were detected, the early/late (CD57+/CD57-) CD8 T effectors ratio was increased at the end of the studied period, as were the percentage of PHA-responding (CD69+) T cells and PB phagocytizing cells. The most prominent lymphoprolipherative responses were measured upon costimulation with LPS+D and PHA+D on day 21. Electron-microscopic studies demonstrated a significant effect of D on both T and B cell activity. TNF-alpha concentration increased progressively from day 7 till the end of the investigation. Maximal concentrations were observed after stimulation with D and LPS. An increased level of specific salivary and serum antibodies against the components of D was found, with highest levels between days 7 and 21. Specific secretory IgA predominated in saliva as compared to IgM and IgG. Our results demonstrate the stimulating effect of Dentavax on PB lymphocyte functional activity and the specific humoral systemic and mucosal immunity. PMID- 16714224 TI - The neutropenia induced by the thalidomide analogue CC-4047 in patients with multiple myeloma is associated with an increased percentage of neutrophils bearing CD64. AB - A major limitation to the treatment of multiple myeloma by the thalidomide analogue CC-4047 (Actimid) is the development of a severe neutropenia. We investigated the hypothesis that this effect may have been due to CC-4047 enhancing the removal of neutrophils from the circulation by altering the expression of surface adhesion molecules required for endothelial binding, by binding to platelets, or by enhancing apoptosis. Flow cytometric analysis was used to examine the expression of neutrophil surface molecules, platelet binding and apoptosis in whole blood samples from 19 patients with multiple myeloma who were assigned to receive either 1, 2, 5 or 10 mg of CC-4047 every other day (e.o.d.) for 28 days. CC-4047 induced dose-related decreases in neutrophil numbers and increases in the percentage of CD64-positive neutrophils, but had little, or no effect on the expression of CD11b, CD62L or CD162, neutrophil platelet binding, or apoptosis. Relative decreases in the neutrophil count were inversely associated with relative increases in the intensity of CD64 expression on neutrophils (r=- 0.307; p=0.028). Although seven patients developed severe neutropenia, none suffered severe or recurrent bacterial infections. The percentage of CD64-positive neutrophils was still increased in eight patients who continued receiving 1-5 mg CC-4047 e.o.d. for several months afterwards, but neutrophil counts were similar to pre-treatment values. PMID- 16714225 TI - Down-regulation of TRRAP-dependent hTERT and TRRAP-independent CAD activation by Myc/Max contributes to the differentiation of HL60 cells after exposure to DMSO. AB - Myc/Max/Mad often play pivotal roles in the proliferation, apoptosis, differentiation and cell cycle progress of leukemia cells. Myc and Mad are known to be unstable proteins and their expression is tightly regulated throughout cell cycle progression and differentiation. Usually, c-Myc expression is implicated in cell growth and proliferation, and the deregulated expression of c-Myc in both myeloid leukemia cells and normal myeloid cells not only blocks terminal differentiation but also its associated growth arrest. HL60 cells could be induced to differentiate into mature granulocytes by DMSO in vitro, but the mechanism of this effect has not been elucidated clearly. We proposed the hypothesis that down-regulation of c-Myc expression by DMSO contributed to the differentiation of HL60 cells by way of activating target genes hTert and CAD. The results showed that c-Myc expression was down-regulated in differentiated HL60 cells but not in exponentially-growing HL60 cells, without or with the target gene activation of hTert and CAD, respectively. Further study indicated that hTert activation is TRRAP-dependent while CAD activation is TRRAP independent. On the other hand, up-regulation of P(21) and P(27) and down regulation of cyclinA and cyclinE also play important roles in induction of the terminal differentiation of HL60 cells. Our results support the hypothesis that c Myc expression and activation of target genes for hTert and CAD play critical roles in the proliferation of HL60 cells, while down-regulation of c-Myc expression and activation of target genes for hTert and CAD contributed to the terminal differentiation of HL60 cells after exposure to DMSO in vitro. PMID- 16714226 TI - Echinacea alkylamides inhibit interleukin-2 production by Jurkat T cells. AB - Alkylamides present in Echinacea species have reported immunomodulatory actions, yet their direct effects on T lymphocytes, key mediators of antiviral immunity, are poorly understood. We hypothesized that constituents present in ethanolic extracts of Echinacea species exert direct immunomodulatory effects on human Jurkat T cells. Modulation of IL-2 production by submaximally stimulated Jurkat cells was determined in response to treatment with extracts prepared from dried aerial parts of Echinacea purpurea. Cells were treated with the extracts, with alkylamides or caffeic acid derivatives isolated from Echinacea species, or with corresponding ethanol vehicle, in the absence or presence of phytohemagglutinin and phorbal ester. E. purpurea extracted in a solvent mixture of 95:5 ethanol/water dose-dependently inhibited IL-2 production. This IL-2 inhibitory activity correlated with the presence of alkylamides but not caffeic acid derivatives, as determined by high performance liquid chromatography/electrospray ionization-mass spectrometry analysis. Simultaneous measurement of secreted IL-2 by ELISA and cell viability by the XTT assay showed that the 95:5 ethanol/water extract of E. purpurea was both IL-2 suppressive and cytotoxic at 50 and 100 microg/mL. Lower concentrations from 6.25 to 25 microg/mL significantly decreased IL-2 production but not cell viability. Alkylamides at concentrations found in a 50 microg/mL extract decreased IL-2 production by approximately 50%. Two Echinacea-derived alkylamides significantly depressed IL-2 production but not cell viability in a dose-dependent manner. Thus, alkylamides present in E. purpurea suppress the ability of activated Jurkat T cells to produce IL-2 independently of direct, cytotoxic effects. PMID- 16714227 TI - [Clinical practice guideline: 2005 update of recommendations for the management of patients with cutaneous melanoma without distant metastases (summary report)]. AB - CONTEXT: The National French federation of comprehensive cancer centres (FNCLCC) and the French society of dermatology (SFD) initiated together the update of clinical practice guideline for the management of patients with cutaneous melanoma in collaboration with the French national cancer institute and with specialists from French public universities, general hospitals and private clinics. This work is based on the methodology developed in the "Standards, Options and Recommendations" (SOR) project. OBJECTIVES: To update SOR guidelines for the management of patients with cutaneous melanoma previously validated in 1998 and French melanoma consensus conference published by SFD and ANAES in 1995. METHODS: The methodology is based on a literature review and critical appraisal by a multidisciplinary group of experts who define the CPGs according to the definitions of the Standards, Options and Recommendations project. Once the guidelines have been developed, they are reviewed by independent reviewers. RESULTS: This article is a summary version of the updated clinical practice guidelines with algorithms. The main questions addressed by the expert group in this update concerned (1) The new AJCC-UICC classification (2) Excision margins (3) Sentinel node biopsy (4) Adjuvant treatments (5) Initial staging and follow up of operated patients. PMID- 16714228 TI - [Synthesis bulletin of 2005 surveillance. Clinical practice recommendations: the use of PET-FDG in cancers of the breast, ovary and uterus]. PMID- 16714229 TI - Current prospects for adult stem cell-based therapies in ocular repair and regeneration. AB - Recent advances in stem cell biology have led to the exploration of stem cell based therapies to treat a wide range of human diseases. In the ophthalmic field, much hope has been placed on the potential use of these cells to restore sight, particularly in those conditions in which other established treatments have failed and in which visual function has been irreversibly damaged by disease or injury. At present, there are many limitations for the immediate use of embryonic stem cells to treat ocular disease, and as more evidence emerges that adult stem cells are present in the adult human eye, it is clear that these cells may have advantages to develop into feasible therapeutic treatments without the problems associated with embryonic research and immune rejection. Here we discuss the current prospects for the application of various adult ocular stem cells to human therapies for restoration of vision. PMID- 16714230 TI - Corneal epithelial cultures generated from organ-cultured limbal tissue: factors influencing epithelial cell growth. AB - PURPOSE: To explore the in vitro proliferative potential of human limbal epithelial cells after 31 degrees C organ-culture storage and to investigate putative factors influencing it. METHODS: 185 cultures of limbal explants were carried-out either from full-thickness explants (n = 102) or from enzymatically dissociated cells (n = 83) seeded on a feeder layer of human keratocytes. Epithelial outgrowth was assessed by phase contrast microscopy using a computerized image analysis software. Cell phenotype was evaluated by transmission electron microscopy and immunocytology. Univariate and multivariate analysis were performed to determine factors influencing epithelial growth in culture. RESULTS: An epithelial outgrowth of 100 square mm or more was observed in 52% of cultures, (average growth area: 440 +/- 256 mm at three weeks). Corneal epithelial phenotype was confirmed by transmission electron microscopy, and cytokeratin pattern. Cytokeratine 19, deltaNp63, nestin and vimentin positive staining revealed undifferentiated epithelial cells in both explant and cell suspension cultures at three weeks. Short death to cornea retrieval time (p < 0.03) and female donors (p < 0.01) were associated with higher cell growth. Enzymatic treatment of explants by trypsin, but not dispase, decreased cell proliferation at two (p < 0.03) and three weeks (p < 0.04). Donor age, duration of corneal storage, and source of the explant did not influence the cell growth. CONCLUSION: Organ-culture conditions can preserve limbal cell mitotic potential if limbal tissue is excised early after circulatory arrest. Human keratocytes can be used as a feeder layer allowing epithelial cells to maintain poorly differentiated phenotype in culture. Further investigations are needed to explain the influence of the donor sex on epithelial cell growth in culture. PMID- 16714231 TI - Pupil response to color signals in cone-contrast space. AB - PURPOSE: It is widely accepted that pupil responses to visual stimuli are determined by the ambient illuminance, and recently it has been shown that changes in stimulus color also contributes to a pupillary control mechanism. However, the role of pupillary responses to chromatic stimuli is not clear. The aim of this study was to investigate how color and luminance signals contribute to the pupillary control mechanism. METHODS: We measured pupillary iso-response contours in M-and L-cone contrast space. The iso-response contours in cone contrast space have been determined to examine what mechanisms contribute to the pupillary pathway. The shapes of the iso-response contour change when different mechanisms determine the response. RESULTS: It was shown that for all subjects, the pupillary iso-response contours form an ellipse with positive slope in cone contrast space, indicating that the sensitivities to the chromatic stimuli are higher than those for the luminance stimuli. The pupil responds maximally to a grating that has a stronger L-cone modulation than the red-green isoluminant grating. CONCLUSIONS: The sensitivity of the chromatic pathway, in terms of pupillary response, is three times larger than that of the luminance pathway, a property that might have utility in clinical applications. PMID- 16714232 TI - The effect of phacoemulsification cataract surgery on the measurement of retinal nerve fiber layer thickness using optical coherence tomography. AB - PURPOSE: To evaluate whether the measurement of retinal nerve fiber layer (RNFL) thickness using optical coherence tomography (OCT) changes after cataract surgery. METHODS: This prospective, randomized, observational, cross-sectional clinical study included 24 eyes of 24 patients who underwent phacoemusification with implant. All patients had no preexisting retinal or optic nerve pathology or other media opacities that might influence the RNFL thickness. The classification and the grading of cataract were based on the Lens Opacities Classification System III (LOCS III). All eyes were scanned immediately before cataract surgery with OCT using Fast RNFL program. All eyes had circular scans around the optic disk with a diameter of 3.4 mm. The OCT was repeated 4 weeks after surgery when the patients returned for postoperative check using the same programme. RESULTS: The mean preoperative RNFL thickness was 84.9 +/- 16.5. The postoperative mean RNFL thickness was 93.0 +/- 17.6. The pre-and postoperative values of the mean RNFL and signal/noise ratio (SNR) are statistically significant (P < 0.05). CONCLUSIONS: Lens opacities may affect the image quality of OCT scans used to measure RNFL thickness as indicated by preoperative low SNR. Cataract extraction results in an apparent increase of the RNFL thickness. PMID- 16714234 TI - Effect of acidosis on isolated porcine retinal vessels. AB - PURPOSE: To study the effect of normocapnic (NA) and hypercapnic acidosis (HA) on the tone, the intracellular calcium level ([Ca(2 +)](i)), and the membrane potential of smooth muscle cells in porcine retinal arterioles. METHODS: Twenty four porcine retinal arterioles were mounted in a wire myograph for isometric recording of the wall tension. The vessels were precontracted with 0.3 microM U46619 and were exposed to NA (pH = 7.0) and HA (pH = 7.0). Intracellular calcium was measured using the fluorophore Fura-2AM (n = 12). In six vessels, 0.1 mM NG nitroarginine methyl ester (L-NAME) was added to block NO synthesis. The membrane potential of smooth muscles cells was measured in situ with sharp glass electrodes (n = 12). RESULTS: NA and HA induced both a decrease in wall tension from 1.04 +/- 0.06 N/m to 0.65 +/- 0.1 N/m (p < 0.01) (NA) and 0.56 +/- 0.1 N/m (p < 0.01) (HA) and a decrease in [Ca(2 +)](i) as evidenced from the change in the Fura-2 fluorescence emission ratio from 0.66 +/- 0.03 to 0.57 +/- 0.05 (p = 0.005) (NA) and 0.56 +/- 0.05 (p = 0.002) (HA). These results were unaffected by inhibition of NO-synthesis. NA and HA also both induced hyperpolarization of the smooth muscle membrane from -18 +/- 0.7 mV during precontraction to -26 +/- 1.9 mV (p = 0.002) (NA) and -24 +/- 2.6 mV (p = 0.02) (HA). CONCLUSIONS: Acidosis induced relaxation of the tone in preconstricted isolated porcine retinal arterioles is associated with a decrease in intracellular calcium and a hyperpolarization of the smooth muscle cells. The acidosis-induced relaxation is independent of CO(2) and is not mediated through NO. PMID- 16714233 TI - Evaluation of an ex vivo model implication for carrier-mediated retinal drug delivery. AB - PURPOSE: The purpose of this study was to evaluate the implication of an ex vivo model for carrier-mediated retinal drug delivery using an Ussing chamber system. METHODS: Dutch Belted Pigmented rabbits weighing 2-2.5 kg were used in these studies. Excised posterior segment tissues (RPE-choroid-sclera and sclera), mounted on the Ussing chamber, were used as an ex vivo model. Transport studies were carried out across sclera and RPE-choroid-sclera (RCS) tissue preparations in the sclera to retina (S --> R) and retina to sclera (R --> S) directions for 3 hr at 37 degrees C. The model was validated by permeability studies with paracellular and transcellular markers ([(3)H]mannitol and [(14)C]diazepam, respectively), tissue viability studies (bioelectrical and biochemical assays), and tissue histology and electron microscopy studies. Functional presence of a carrier-mediated transport system for folic acid (folate receptor alpha) was investigated on the basolateral side of the rabbit retina. RESULTS: Results from bioelectrical, biochemical, and histological evaluation of tissue provide evidence that the RCS tissue preparation remains viable during the period of transport study. Permeability values of [(3)H]mannitol across sclera were 4.18 +/ 1.09 x 10(- 5) cm/s (R --> S) and 4.11 +/- 1.09 x 10(- 5) cm/s (S --> R) and across RCS were 1.77 +/- 0.31 x 10(- 5) cm/s (S --> R) and 1.60 +/- 0.19 x 10(- 5) cm/s (R --> S). Permeability values of [(14)C]diazepam across sclera were 2.37 +/- 0.38 x 10(- 5) cm/s (R --> S) and 2.70 +/- 0.70 x 10(- 5) cm/s (S --> R) and across RCS were 3.12 +/- 0.12 x 10(- 5) cm/s (R --> S) and 2.77 +/- 0.25 x 10(- 5)cm/s (S --> R). The rate of [(3)H]folic acid transport across RCS was found to be significantly higher in the S -->R direction (16.34 +/- 0.94 fmoles min(-1) cm(-2)) as compared with R --> S direction (9.38 +/- 1.44 fmoles min(-1) cm(-2)) and nearly 10-fold higher across sclera as compared with RCS in both directions. Transport of [(3)H]folic acid was found to be pH and temperature dependent and was inhibited by 44.5%, 35.1%, and 50.3% in the presence of unlabeled folic acid, 5-methyltetrahydrofolate (MTF), and Methotrexate (MTX). CONCLUSIONS: The RCS tissue preparation mounted on the Ussing chamber system, an ex vivo model, can be a useful tool for identification and characterization of carrier-mediated systems present on RPE (a major barrier for retinal drug delivery) and to study carrier mediated retinal drug delivery via prodrug derivatization. PMID- 16714235 TI - Toxicity of subretinal ribozyme to the proliferating cell nuclear antigen and 5 fluorouracil in rat eyes. AB - PURPOSE: To investigate the subretinal toxicity profile of the ribozyme to the proliferating cell nuclear antigen (PCNA-Rz) and 5-fluorouracil (5-FU), as well as the highest nontoxic subretinal dose of the mixture of the two agents in rat eyes. METHODS: Brown-Norway rats received subretinal injections of 1 microg, 10 microg, and 100 microg/microl PCNA-Rz and 0.06 microg/microl, 0.3 microg/microl, and 1.5 microg/microl 5-FU in the right eyes, and the left eyes were injected with H-BSS as control. Each dose was tested on 5 eyes in a 5 microl volume. In a second study, a combination of 5-FU (1.5 microg/microL) with varying 10-30-50 microg/microl doses of PCNA-Rz was tested in a regimen of four sequential subretinal injections. Toxicity was monitored by biomicroscopy, indirect ophthalmoscopy, electroretinography (ERG), and histology. RESULTS: The highest nontoxic dose for subretinal PCNA-Rz was 10 microg/microl, whereas 100 microg/microl showed disturbance of pigmentation with corresponding histological changes of retinal photoreceptor loss and retinal pigment epithelium proliferation or irregularities. Subretinal injection of all three doses of 5-FU did not show any toxicity. Serial injections of a mixture of 1.5 microg/microl 5 FU with 10 microg/microl of PCNA-Rz was found to be safe in rat eyes. CONCLUSIONS: Subretinal injections of the combination of PCNA-Rz (10 microg/microl) and 5-FU (1.5 microg/microl) demonstrated to be safe in rat eyes during the course of this study, even with a multiple administration of four injections. PMID- 16714236 TI - Effect of H-7 and Lat-B on retinal physiology. AB - PURPOSE: To investigate the effects of H-7 and Latrunculin B (Lat-B) on retinal vascular permeability and electrophysiology at concentrations that increase outflow facility in monkeys. METHODS: One eye of 1 rhesus and 22 cynomolgus monkeys received an intravitreal bolus injection of H-7 or Lat-B; the opposite eye received vehicle. Multifocal electroretinograms (mfERGs), and photopic and scotopic full-field electroretinograms (ffERGs, sERGs) were recorded in subsets of monkeys at baseline and at multiple time-points post-H-7 or Lat-B. Vitreous fluorophotometry (VF) and fluorescein angiography (FA) were also performed. RESULTS: No differences between the H-7 or Lat-B treated and control eyes were found in ffERGs, mfERGs, sERGs, or in FAs in any monkey. No significant difference was found in vitreous fluorescein levels between H-7 treated or Lat-B treated vs. control eyes. CONCLUSIONS: No effect on retinal vascular permeability or retinal electrophysiology was apparent after intravitreal administration of H 7 or Lat-B at doses that increase outflow facility and lower IOP when given intracamerally. PMID- 16714237 TI - Identification of a KRAB-zinc finger protein binding to the Rpe65 gene promoter. AB - PURPOSE: We wish to identify transcriptional factors involved in regulation binding to the proximal promoter region of the RPE65 gene that confers RPE specific expression. METHODS: We incubated human D407 RPE cell nuclear extract with double-stranded (sense 5-prime biotinylated) oligonucleotides, based on the RPE65 proximal gene promoter, bound to streptavidin-Dynabeads. Bound nuclear proteins were eluted, separated on SDS-PAGE, and analyzed by mass spectrometry. Peptide sequence was used to identify cDNA clones that were subcloned into pCDNA3.1 for expression and co-transfection into D407 cells to assess transcriptional activation of mouse Rpe65 gene promoter/reporter constructs. SiRNA interference was used to suppress ZNF492 expression. RESULTS: We identified a D407 nuclear protein binding to biotinylated-DNA/streptavidin beads as the product of clone KIAA1473 encoding a protein named ZNF492. ZNF492 has an open reading frame of 531 amino acids with a truncated N-terminus and lacks the usual Kruppel-associated box-A (KRAB-A) while KRAB-B remains intact and has 12 C2H2 zinc-fingers in tandem arrangement. Co-expression in D407 cells of ZNF492 protein did not activate TR1, a mouse Rpe65 gene promoter/reporter construct with 49-bp 5 prime flanking sequence, but did activate construct TR2, containing 188-bp 5 prime flanking sequence, by 2.5-fold, and the longer constructs TR4, containing 655-bp 5-prime flanking sequence, and TR5, containing 1240-bp 5-prime flanking sequence, by about 2-fold. SiRNA-mediated suppression of ZNF492 in D407 resulted in decreased Rpe65 promoter activity. CONCLUSIONS: We have identified ZNF492, a KRAB-zinc finger protein, by its interaction with immobilized RPE65 promoter DNA sequence. This KRAB-zinc finger protein serves as a moderate transcriptional factor for Rpe65 gene upregulation. In ZNF492, absence of KRAB-A might reduce or prevent co-repressor binding to account for the modest upregulation of Rpe65 gene expression. PMID- 16714238 TI - Roles of adherence and matrix metalloproteinases in growth patterns of fungal pathogens in cornea. PMID- 16714240 TI - Preserved cognition and functional independence after a large right posterior cerebral artery infarct: longitudinal clinical and neuropathological findings. AB - BVR was 77 years old when he sustained a large posterior cerebral artery territory infarct. Medical, cognitive and functional data collected on four occasions over 10 years initially revealed circumscribed neurological signs, no functional or cognitive deficits. BVR became significantly impaired only after two other strokes, 3 years before death. On brain MRI, the lesions involved large portions of the right occipital and temporal cortices, the right thalamus, and the left cerebellum, as well as thinning of the corpus callosum. Postmortem investigations revealed additional recent vascular lesions in the occipital region. This case study underscores the importance of comprehensive assessment methods combining neurological, neuroimaging and cognitive tools. PMID- 16714241 TI - Qualitatively different semantic representations for abstract and concrete words: further evidence from the semantic reading errors of deep dyslexic patients. AB - This paper presents an investigation of the hypothesis that conceptual knowledge for abstract and concrete items is underpinned by qualitatively different representational frameworks (Crutch and Warrington, 2005a). A re-analysis of the semantic reading errors of four deep dyslexic patients is presented, examining the incidence of semantically associated and semantically similar errors in response to abstract and concrete target words. The results demonstrate that abstract target words elicit a greater proportion of associative than similar errors, while concrete words show the reverse pattern. These findings provide evidence which converges with that previously documented for a semantic refractory access dysphasic to suggest that abstract concepts are represented in an associative network while concrete concepts are represented in a categorical framework. PMID- 16714242 TI - A mirror in the mind: a case of visual allaesthesia in homonymous hemianopia. AB - We report the case of a 61-year-old female patient with intracranial bleeding in left parietal and parieto-temporal regions and a history of epilepsy and migraine. MR images showed lesions of the optic radiation, but primary visual cortical areas were intact. Perimetric testing revealed an incomplete right hemianopia. The patient claimed that visual percepts from her intact field were projected as "mirror images" into the hemianopic field. The illusory images were weak and sometimes difficult to detect, but focusing spatial attention on the "mirror" image increased its saliency. Drawings the patient made of her pseudo hallucinations revealed that the illusions were lateral transpositions instead of mirror images of real objects. The illusions were tilted in clockwise direction and were never colored, although color discrimination was unimpaired in the patient's left hemifield. We quantified the characteristics of the pseudohallucinations in several experiments: The patient was asked to adjust the position, rotation angle, and size of a white test card in her blind field so that it corresponded with the illusory projection of a card of standard size and position that was presented in the intact field. The test card was compressed in horizontal size by 20% and positioned 17 degrees visual angle to the right of the standard, shifted 1.5 degrees upward or downward, and rotated in clockwise direction by 22.6 degrees on average. Large objects in the intact field were projected incompletely into the blind area. Our patient's symptoms are similar to those reported in earlier case studies of visual allaesthesia. We hypothesize that the "mirror image" is induced by sparse input from contralesional V1 via the corpus callosum upstream of the lesion site and a lack of inhibition or hyperexcitability of ipsilesional early visual areas after deafferentation. The rotation of the illusions may be induced by the parietal lesions causing faulty co-ordinate computations, e.g., an inability to integrate visual and otholitic input. PMID- 16714243 TI - Complex visual hallucinations as post-ictal cortical release phenomena. AB - Visual hallucinations can be elementary or complex and appear in a wide variety of neurological disorders. Hallucinations associated with seizures can either be ictal or part of a post-ictal cortical release phenomenon, and their features may help distinguish their etiology. We describe two patients with suspected ictal occipital lobe lesions with elementary hallucinations that were followed by post ictal complex hallucinations.The pathophysiologies of elementary and complex hallucinations are reviewed. The sequential occurrence of these two distinct types of hallucinations in these patients emphasizes the observation that hallucinations associated with seizures can result from both irritative and cortical release mechanisms. PMID- 16714244 TI - Do visual attentional factors contribute to phonological ability? studies in adult dyslexia. AB - A case study approach was taken to examine the role of visual attention and auditory memory processes in eight adults with dyslexia, all of whom demonstrated phonological difficulties. The participants were administered a battery of neuropsychological tests and participated in an attentional cueing experiment. Individual data revealed that, although one adult with dyslexia showed overt visual attention deficits on a visual search task, and five showed auditory working memory deficits, the difficulty that all of the adults with dyslexia had in common was with covert shifts of attention toward and away from fixation. These results indicate that deficits in overt visual attentional processing and working memory can be present with dyslexia, but neither is a necessary requirement. Overall, the results suggest that covert visual attention makes a significant contribution to phonological ability, which thus has implications for reading ability. PMID- 16714245 TI - Preservation of Emmert's law in a visual form agnosic. AB - Size constancy was investigated in DF, a patient with visual form agnosia, using a technique based on Emmert's law of visual after-images. DF was first given a task in which she was asked to indicate the distance of a vertical surface and a task where she had to estimate the width of a series of squares (widths ranging from 5 cm to 35 cm) placed at varying distances and having a constant visual angle. In the distance estimation task, DF greatly overestimated the distance of the vertical surface placed in front of her. DF also had great difficulty performing the size estimation task. DF then performed a task in which she stared at a bright 5 cm square for a brief period of time at a distance of 30 cm followed by the presentation of a vertical surface which varied in distance and was asked to indicate the width of the after-image either verbally or manually. DF's after-images conformed to the size-distance relationship predicted by Emmert's law--as the distance of the vertical surface increased her perception of the size of the after-images also increased. These data demonstrate that although DF is rather impaired in tasks that require explicit estimates of size and distance, at some level, DF must have relatively intact size constancy mechanisms given that her estimates of the width of the after-image conform to Emmert's law. Thus, the processes underlying explicit judgements of size and distance appear to differ from those underlying the size and distance scaling of after-images. PMID- 16714246 TI - Aldosterone synthase (CYP11B2)-344T/C polymorphism and renoprotective response to losartan treatment in diabetic nephropathy. AB - OBJECTIVE: It has been suggested that an aldosterone synthase gene polymorphism (CYP11B2 -344T/C) is predictive of the blood pressure lowering effect of angiotensin II receptor blockers in essential hypertension. We investigated whether this polymorphism is predictive of reductions in blood pressure and albuminuria and preservation of glomerular filtration rate (GFR) during short term and long-term treatment with losartan in 57 hypertensive type-1 diabetic patients with diabetic nephropathy. MATERIAL AND METHODS: After a 4-week washout period, patients received losartan (100 mg o.d.) and were followed for a mean follow-up of 36 months. At baseline, after 2 and 4 months, and every 6 months thereafter, GFR (51Cr-EDTA-clearance), albuminuria and 24-h blood pressure were determined. The CYP11B2 -344T/C polymorphism was determined by standard polymerase chain reaction (PCR). RESULTS: The TT, CT and CC genotypes were found in 28 %, 58 % and 14 % of patients, respectively. At baseline albuminuria and blood pressure did not differ between genotype groups. Plasma aldosterone levels (geometric mean (95 % CI)) were similar at baseline: 87 (60-125), 77 (53-112), and 89 (49-161) pg mL(-1) and during follow-up (not significant). After initiation of losartan treatment, comparable mean (SE) reductions in blood pressure and albuminuria were seen in patients with TT, CT and CC genotypes (p >0.6 between groups). After long-term follow-up, there was a tendency towards a difference in systolic blood pressure reduction (p = 0.07, one-way ANOVA), suggesting a poorer response in patients with the CC genotype. No significant difference in rate of decline in GFR (median (range)) was seen between groups (TT, CT, CC): 4.2 (-1.0 to 16.0), 3.2 (-1.6 to 13.8) and 2.6 (-0.1 to 11.0) mL min(-1)year(-1), respectively (p = 0.5). CONCLUSIONS: Compared to a previous smaller study of angiotensin II receptor blockade in essential hypertension, we could not confirm that CYP11B2 -344T/C genotypes contribute towards explaining the observed variability in response to treatment with angiotensin II receptor blockers, which could be due to lack of power. PMID- 16714247 TI - Determination of melatonin in saliva using automated solid-phase extraction, high performance liquid chromatography and fluorescence detection. AB - A sensitive bioanalytical method for the determination of melatonin in saliva by solid-phase extraction (SPE), high-performance liquid chromatography (HPLC) and fluorescence detection has been developed and validated. Saliva was collected with a Salivette sampling device (Sarstedt) and a mixed-mode SPE column was used for the extraction of melatonin and internal standard (N-acetyl-6 methoxytryptamine) from the saliva. Chromatographic separation was performed using a HyPurity C18 LC column (150 x 2.1 mm) with mobile phase acetonitrile ammonium hydrogen carbonate buffer, 0.015 M, pH 6.8 (23:77, v/v). Excitation and emission wavelengths were set to 285 nm and 345 nm, respectively. The within-day precision for the method at 50 pmol/L was 7.9 % and the between-day precision was 10.5 %. The limit of quantification was 50 pmol/L. PMID- 16714248 TI - Determinants of risk factors of atherosclerosis in the postinfarction period: the Tallinn MI study. AB - Environmental as well as genetic factors are involved in the pathogenesis of myocardial infarction. The disease is a frequent cause of mortality in the middle aged male population of Estonia. The high prevalence of premature myocardial infarction (PMI) in this country is not fully understood. The association of atherogenic and thrombogenetic risk factors with lifestyle was evaluated in men who had suffered myocardial infarction at 55 years of age (n = 71) and in randomly selected corresponding controls (n = 85). Serum routine lipids, apolipoprotein (apo)A-I, apoB, apoE polymorphism, lipoprotein(a) and fibrinogen levels were determined. Behavioural risk factors, indices of obesity, blood pressure and pedigree data were registered. In 80.6 % of PMI subjects some type of hyperlipidaemia was observed (European Atherosclerosis Society Classification) and lipid-lowering drugs were taken by 13.9 % of patients. In PMI patients the most common positive determinants of atherogenic lipoprotein indices were waist to-hip ratio and physical inactivity, and in controls, waist-to-hip ratio and apoE phenotype. The odds ratio (OR) of PMI was 8.9-fold greater in the highest tertile of apoB/apoA-I distribution compared with the lowest tertile. The OR of PMI in the highest tertile of fibrinogen distribution versus the lowest tertile was 6.2 (95 % CI 2.46-15.44), and OR of PMI in the highest Lp(a) tertile versus the lowest was 3.1 (95 % CI 1.31-7.40). Thus, atherogenic dyslipidaemia was the most serious cardiovascular risk factor among PMI patients. From two thrombogenesis-related markers, the levels of fibrinogen and Lp(a), the first one was more strongly associated with PMI status. PMID- 16714249 TI - Nitric oxide induction as a novel immunoepidemiological target in malaria infected patients from endemic areas of the Islamic Republic of Iran. AB - OBJECTIVE: Malaria has been prevalent for a long time in Iran and continues to be a health problem despite substantial control programs. In addition to numerous cytokines, nitric oxide (NO) is thought to be a key molecule and a novel target of malaria immunopathology. MATERIAL AND METHODS: The objective of this research was to measure reactive nitrogen intermediates (RNI) as stable metabolites of NO induction in plasma of malaria-infected patients in Iran. In this study, 235 blood samples from malaria patients and 80 blood samples from healthy controls were randomly collected from different malarial endemic provinces of Iran, located in southeastern (Sistan & Balouchestan, Hormozgan, Kerman) and northwestern (Ardabil) areas. The involvement of NO in malaria patients has been investigated by statistical analysis of RNI values. Griess micro assay (GMA) was used during Plasmodium vivax, P. falciparum and mixed infections, in order to evaluate whether RNI changes are related to the provincial areas, parasite strains, clinical symptoms and age and gender parameters. RESULTS: The results showed a significant increase of RNI level in malaria patients compared with the control groups of Ardabil (p<0.01), Sistan & Balouchestan, Hormozgan and Kerman (p<0.001) provinces. The level of RNI was higher in mixed plasmodial infection than in single infection. CONCLUSIONS: The high level of RNI was dependent on the type of infection, the plasmodia strain, the clinical symptoms, the age groups and the endemic provinces. Although, this study did not clarify the pathogenic and/or protective role of NO in malaria, our findings provide a novel immunoepidemiological aspect of basal NO production in patients with malaria in endemic areas in Iran. PMID- 16714250 TI - Lactate elimination and glycogen resynthesis after intense bicycling. AB - OBJECTIVE: Muscles break down glycogen to lactate during intense exercise, and in the recovery period, glycogen reappears while lactate disappears. The purpose of this study was to examine to what extent lactate is resynthesized to glycogen within the formerly active muscles themselves in man. MATERIAL AND METHODS: Fifteen healthy young men cycled for 2 min to exhaustion. Muscle biopsies were taken from the knee extensor muscle before the exercise, just after the ride, and again after 45 min of recovery. In addition, blood samples were taken from the femoral artery and vein, and the leg blood flow was measured using the ultrasound Doppler technique. The muscle biopsies were analysed for glycogen, lactate and other metabolites, and the blood samples were analysed for lactate and glucose. The exchanges of lactate and glucose of the leg were assessed by multiplying the measured arterio-venous (a-v) differences by the blood flow. RESULTS: During the exercise the muscles broke down 20+/-4 mmol glycogen kg(-1) wet muscle mass and produced 26+/-1 mmol lactate kg(-1). In the recovery period after 24+/-1 mmol lactate kg(-1) had disappeared, of which 48 % was released to the blood, 52 % disappeared within the muscle. An R-value of 0.62 across the leg suggests that none of the lactate was oxidized. Altogether, 10+/-3 mmol glycogen kg(-1) reappeared during recovery. Glucose uptake accounted for 2 mmol kg(-1) and glycolytic intermediates (G-6-P and free glucose) accounted for 4 mmol kg(-1); 4 mmol glycogen kg(-1) (42 %) reappeared from unknown sources. CONCLUSIONS: The present data are compatible with the idea that around half of the lactate produced during intense bicycling is resynthesized to glycogen within the working muscles themselves in the recovery period after the bicycling. PMID- 16714251 TI - Vitamin D and its binding protein Gc: long-term variability in peri- and postmenopausal women with and without hormone replacement therapy. AB - OBJECTIVE: Measurement of plasma 25-hydroxyvitamin D (25OHD) level is often used to evaluate a patient's vitamin D status. The purpose of this study was to investigate the variability in individual plasma 25OHD- and vitamin D-binding protein- (Gc) levels over a 5-year period in postmenopausal women with and without hormone replacement therapy (HRT). MATERIAL AND METHODS: A total of 187 women were followed-up for 5 years. At baseline, 89 women were allocated to treatment with HRT, given orally. Measurements were performed at baseline and after 1, 2 and 5 years of follow-up. RESULTS: At baseline, 25OHD levels were positively associated with sunbathing and use of vitamin D supplements, and inversely associated with smoking. HRT therapy increased plasma levels of Gc (+8 %) but did not affect 25OHD levels or the free 25OHD index (molar ratio of 25OHD- to Gc levels). Among those classified in the lowest 25OHD tertile at baseline, 40 % remained in the lowest tertile during all subsequent measurement time-points. Similarly, 32 % of those classified in the highest baseline tertile remained in the highest tertile during all subsequent measurements. Use of the free 25OHD index showed similar results. No independent predictors of changes in vitamin D tertiles during follow-up were identified, which suggests that the observed variation was caused by the intra-individual variation in measured parameters. For all participants, the within-patient variability in 25OHD measurements was between 13 % and 19 %. CONCLUSIONS: In healthy postmenopausal women, HRT increases Gc levels. Owing to the high intra-individual variation in plasma 25OHD, it seems questionable to use a single estimate as a predictor of individual vitamin D status. PMID- 16714252 TI - Plasma antioxidant capacity among middle-aged men: the contribution of uric acid. AB - OBJECTIVE: Although assays of plasma antioxidant capacity encompass interactions between various antioxidants, uric acid concentration can exert a predominant effect on results. Therefore, individual differences in uric acid concentration may explain a many of the differences in antioxidant capacity. The objective of this study was to measure the antioxidant capacity of plasma samples with and without uric acid in order to provide more information about how the concept of antioxidant capacity could be applied. MATERIAL AND METHODS: Antioxidant capacity was measured using an enhanced chemiluminescence assay, and uric acid was removed from the samples using uricase. RESULTS: Antioxidant capacity was positively correlated with uric acid concentration, body mass index, waist circumference, abdominal sagittal diameter and the concentrations of insulin and triglycerides. These correlations were not evident when uric acid was eliminated from the sample, but antioxidant capacity was correlated with lipid concentration; this may partly reflect tocopherols that are transported by lipid molecules. CONCLUSIONS: The significance of the contribution of uric acid to the antioxidant capacity could differ according to the type of study. Antioxidant capacity measurements in cross-sectional studies may be presented both with and without the contribution of uric acid, because the absence of such data complicates interpretation of results when different populations are compared. PMID- 16714253 TI - Improvement of real-time polymerase chain reaction for quantifying TNF-alpha mRNA expression in inflamed colorectal mucosa: an approach to optimize procedures for clinical use. AB - OBJECTIVE: The precise measurement of local tumor necrosis factor alpha (TNF alpha) expression in tissue is important in understanding the pathogenesis of inflammatory bowel diseases (IBD). Real-time polymerase chain reaction (PCR) is a sensitive, versatile method and is becoming a commonly used tool for the quantification of gene expression. The aim of this study was to optimize the laboratory procedure for biopsy sampling, storage and calibration of result for TNF-alpha mRNA quantification with real-time PCR of colorectal biopsies. MATERIAL AND METHODS: Endoscopic biopsies from the colorectum were obtained from 18 patients with ulcerative colitis (UC), 11 patients with Crohn's disease (CD) and 18 normal controls. Optimization of procedures for real-time PCR performance was carried out. RESULTS: The transport medium, RNAlater, exhibited a high preservation effect against RNA degradation even after 8 days of storage at room temperature; one biopsy from each patient was sufficient for RNA extraction, cDNA synthesis and TNF-mRNA quantification. An assay was established with a technical reproducible sensitivity of 100 copies/microL. The observed interassay variations were 7.4 % coefficient of variation (CV) and 7.2 % CV in low and high TNF-alpha mRNA expression biopsies, respectively. TNF-alpha mRNA levels in colorectal biopsies from patients with either CD or moderate to severe UC were markedly increased, and 8 approximately 9-fold higher than those in healthy controls. CONCLUSIONS: This optimization improves the clinical use of real-time PCR for quantification of TNF-alpha gene expression in colorectal biopsies and provides a sensitive reproducible assay. PMID- 16714254 TI - Vascular endothelial growth factor serum concentrations in hypercholesterolemic patients. AB - OBJECTIVE: Vascular endothelial growth factor (VEGF) promotes normal and pathological angiogenesis. VEGF is a chemotactic factor for macrophages and vascular smooth muscle cells, and induces synthesis of metalloproteinases and adhesion molecules. VEGF expression is regulated by hypoxia, cytokines, oncogenes, and oxidized low-density lipoprotein (LDL). The purpose of this study was to determine the relationship between levels of lipid parameters and VEGF, to investigate whether pravastatin treatment influences VEGF serum concentrations, and to examine the relationship between VEGF and the variations in post-treatment lipid and inflammatory parameters. MATERIAL AND METHODS: Eighteen patients aged 48+/-6.8 years with total cholesterol (TC) >6.1 mmol/L comprised the hypercholesterolemic group. The controls included 12 individuals aged 50+/-7.4 years with TC <5.1 mmol/L. TC, high-density lipoprotein cholesterol (HDLC), triglycerides, LDLC, C-reactive protein (CRP), and VEGF were determined in both groups at baseline, and in the hypercholesterolemic group after 4 months of treatment with 20 mg/day pravastatin. RESULTS: A significant correlation was observed between concentrations of VEGF and TC, LDLC and TG, and a significant difference in VEGF concentration was observed between the control group (mean 142 ng/L) and the hypercholesterolemic group (mean 272.9 ng/L). A significant decrease was observed in TC (14.7 %), LDLC (21.5 %), CRP (22.7 %), and VEGF (14.8 %) after 4 months of treatment with pravastatin. CONCLUSIONS: A relationship was found between serum levels of VEGF and most atherogenic lipoproteins. In patients with hypercholesterolemia treated with pravastatin, a reduction in VEGF and CRP was seen in addition to lipid decreases. PMID- 16714255 TI - Erosion of the integrity of public health science in the USA. PMID- 16714256 TI - Socioeconomic differences in severe back morbidity. PMID- 16714257 TI - How much is the brain involved in glaucoma? PMID- 16714258 TI - Anatomical diagnosis. PMID- 16714259 TI - Pterygium. PMID- 16714260 TI - Endoscopic and transscleral cyclophotocoagulation. PMID- 16714261 TI - Primary care ophthalmology. PMID- 16714262 TI - The timing of patching treatment and a child's wellbeing. PMID- 16714263 TI - Clinical characterisation of a family with retinal dystrophy caused by mutation in the Mertk gene. AB - BACKGROUND/AIM: MERTK, a tyrosine kinase receptor protein expressed by the retinal pigment epithelium (RPE), is mutated in both rodent models and humans affected by retinal disease. This study reports a survey of families for Mertk mutations and describes the phenotype exhibited by one family. METHODS: 96 probands with retinal dystrophy, consistent with autosomal recessive segregation, were screened by direct sequencing. A family homozygous for a likely null allele was investigated clinically. RESULTS: A novel frame shifting deletion was identified in one of 96 probands. Other polymorphisms were detected. The deletion allele occurred on both chromosomes of four affected family members. Electrophysiology demonstrated early loss of scotopic and macular function with later loss of photopic function. Visual acuities and visual fields were preserved into the second decade. Perception of light vision was present in a patient in the fourth decade. A "bull's eye" appearance and a hyperautofluorescent lesion at the central macula were consistent clinical findings. CONCLUSIONS: Mutations in Mertk are a rare cause of ARRP in humans. The study extends the phenotypic characteristics of this retinal dystrophy and shows distinctive clinical signs that may improve its clinical identification. The moderate severity and presence of autofluorescence implies that outer segment phagocytosis is not entirely absent. PMID- 16714264 TI - Penetrating keratoplasty: outcomes from a corneal unit compared to national data. AB - AIMS: To determine long term graft survival rates and visual results for different indications for penetrating keratoplasty from a single institution over 10 years and compare these to national outcome data. METHODS: Retrospective chart analysis. 784 records were available for review of 1096 consecutive penetrating keratoplasty procedures performed between 1990 and 1999 (72%). Outcomes of graft survival, visual acuity, and astigmatism were analysed and compared to national outcome data supplied by the UK Transplant Service. RESULTS: At 5 year follow up, overall graft survival was 66%. This was subdivided into 98% for keratoconus, 86% for viral keratitis, 85% for Fuchs' dystrophy, 84% for pseudophakic bullous keratopathy, 55% for regrafts, and 57% for other diagnoses. There was a significantly higher graft survival rate for all diagnostic subgroups except Fuchs' dystrophy at 3 years of follow up compared to the national average. Best corrected visual acuity at 5 years was 6/18 or better in 53% of cases. The mean keratometric astigmatism was 3.4 dioptres. CONCLUSION: Penetrating keratoplasty is a safe and effective treatment for selected corneal disorders. Penetrating keratoplasty for viral keratitis may achieve good results with long term antiviral treatment. Patients may achieve better outcomes if their surgery is performed at specialist centres. PMID- 16714265 TI - Transscleral diode laser cyclophotocoagulation as primary surgical treatment for secondary glaucoma in juvenile idiopathic arthritis: high failure rate after short term follow up. AB - AIM: To evaluate the success rates of transscleral diode cyclophotocoagulation (TD-CPC) for secondary glaucoma in patients with juvenile idiopathic arthritis (JIA) as primary surgical treatment. METHODS: Retrospective chart review of 12 paediatric patients with JIA associated uveitis and secondary open angle glaucoma. RESULTS: 21 eyes of the 12 patients had chronic anterior uveitis, 21 eyes had secondary open angle glaucoma, and 19 eyes were treated 41 times with TD CPC. Patients underwent a mean of 2.15 treatment sessions per eye. Pretreatment intraocular pressure (IOP) was 30.2 (5.5) mm Hg before the first treatment, 30.5 (5.7) mm Hg before the second treatment, and 28.7 (6.3) mm Hg before the third treatment. Reduction of IOP 6 weeks after treatment was highest after the second and third treatments with 9.4 (8.8) and 8.7 (5.8) mm Hg, respectively and 5.2 (10.7) mm Hg after the first treatment. Qualified success (IOP 1 and 14 years. About 15.8% of the patients presented with status epilepticus during the course of disease. About 17% of the patients showed transient responsiveness (> or =1 year seizure freedom) to antiepileptic drug therapy either after initial therapy (50%) or later in the course of epilepsy (50%). Patients with FCD represent a heterogeneous group. Different age at epilepsy onset and transient responsiveness to antiepileptic drugs in approximately 17% of patients may reflect different dynamics in epileptogenicity of the underlying FCD. Dual pathology may be associated with different pathomechanisms in patients with and without febrile seizures. PMID- 16714317 TI - SERCA1 and calsequestrin storage myopathy: a new surplus protein myopathy. AB - We describe four patients, from four different families, affected by a mild myopathy or asymptomatic elevated serum creatine kinase levels, in whom toluidine blue-stained semithin sections of muscle specimens revealed inclusions of different size and shape. The inclusions did not stain by routine histochemical studies. The sarcoplasmic or endoplasmic reticulum calcium 1 (SERCA1) ATPase and/or calsequestrin reactivity of inclusions, by immunohistochemistry, and the SERCA1- and calsequestrin-increased expression, by immunoblot, suggested that inclusions were constituted by an excess of proteins normally present in the terminal cisternae of sarcoplasmic reticulum. Our cases, both sporadic and familial, represent a new type of surplus protein myopathy. PMID- 16714318 TI - MFN2 mutation distribution and genotype/phenotype correlation in Charcot-Marie Tooth type 2. AB - Mutations in mitofusin 2 (MFN2) have been reported in Charcot-Marie-Tooth type 2 (CMT2) families. To study the distribution of mutations in MFN2 we screened 323 families and isolated patients with distinct CMT phenotypes. In 29 probands, we identified 22 distinct MFN2 mutations, and 14 of these mutations have not been reported before. All mutations were located in the cytoplasmic domains of the MFN2 protein. Patients presented with a classical but rather severe CMT phenotype, since 28% of them were wheelchair-dependent. Some had additional features as optic atrophy. Most patients had an early onset and severe disease status, whereas a smaller group experienced a later onset and milder disease course. Electrophysiological data showed in the majority of patients normal to slightly reduced nerve conduction velocities with often severely reduced amplitudes of the compound motor and sensory nerve action potentials. Examination of sural nerve specimens showed loss of large myelinated fibres and degenerative mitochondrial changes. In patients with a documented family history of CMT2 the frequency of MFN2 mutations was 33% indicating that MFN2 mutations are a major cause in this population. PMID- 16714319 TI - Unconscious vision: new insights into the neuronal correlate of blindsight using diffusion tractography. AB - The existence of several types of unconscious vision, or 'blindsight', has convincingly been demonstrated in numerous studies, and their neuronal correlates have been hypothesized according to the nature of the residual vision observed. We used diffusion tensor imaging (DTI) tractography to demonstrate an association between the presence of 'Type I'- blindsight or 'attention blindsight' and reconstructed superior colliculi (SC) fibre tracts in hemispherectomized subjects, in support of the hypothesis that this subcortical structure plays a pivotal role in this type of blindsight. Before the DTI study, 'Type I' blindsight was identified in two of four hemispherectomized subjects by using a spatial summation effect paradigm, an indirect behavioural method, in which subjects were unaware of a stimulus presented in their blind visual field and were required to respond to an identical stimulus presented simultaneously in their intact field. SC tracts were then reconstructed in six control subjects, the two hemispherectomized subjects with blindsight and the two hemispherectomized subjects without blindsight. Whereas control subjects demonstrated mainly ipsilateral connections to visual association areas, parietal cortex, prefrontal areas and to an area close to the frontal eye fields, hemispherectomized subjects with blindsight showed ipsi- and contralateral connections from the SC to visual association areas, primary visual areas, parietal areas, prefrontal areas and to the posterior part of the internal capsule. In contrast, no projections from the SC on the hemispherectomized side were observed in hemispherectomized subjects without blindsight, in support of a key role of this structure in 'Type-I' or 'attention blindsight'. PMID- 16714321 TI - Implications of mechanism-based inhibition of CYP2D6 for the pharmacokinetics and toxicity of MDMA. AB - The aim of this study was to model the in vivo kinetic consequences of mechanism based inhibition (MBI) of CYP2D6 by 3,4 methylenedioxymethamphetamine (MDMA, ecstasy). A model with physiologically-based components of drug metabolism was developed, taking account of change in the hepatic content of active CYP2D6 due to MBI by MDMA. Based on the in vitro information, plasma concentration time profiles of MDMA after various doses were computed and compared with reported observations. The analysis suggested that a typical recreational MDMA dose could inactivate most hepatic CYP2D6 within an hour, and the return to a basal level of CYP2D6 could take at least 10 days. Thus, the genetic polymorphism of CYP2D6 and coadministration of CYP2D6 inhibitors may have less impact on MDMA pharmacokinetics and the risk of acute toxicity than previously thought. This is consistent with clinical observations that indicate no obvious link between inherited CYP2D6 deficiency and acute MDMA intoxication. PMID- 16714323 TI - Effects of psilocybin on time perception and temporal control of behaviour in humans. AB - Hallucinogenic psilocybin is known to alter the subjective experience of time. However, there is no study that systematically investigated objective measures of time perception under psilocybin. Therefore, we studied dose-dependent effects of the serotonin (5-HT)2A/1A receptor agonist psilocybin (4-phosphoryloxy-N, N dimethyltryptamine) on temporal processing, employing tasks of temporal reproduction, sensorimotor synchronization and tapping tempo. To control for cognitive and subjective changes, we assessed spatial working memory and conscious experience. Twelve healthy human volunteers were tested under placebo, medium (115 microg/kg), and high (250 microg/kg) dose conditions, in a double blind experimental design. Psilocybin was found to significantly impair subjects' ability to (1) reproduce interval durations longer than 2.5 sec, (2) to synchronize to inter-beat intervals longer than 2 sec and (3) caused subjects to be slower in their preferred tapping rate. These objective effects on timing performance were accompanied by working-memory deficits and subjective changes in conscious state, namely increased reports of 'depersonalization' and 'derealization' phenomena including disturbances in subjective 'time sense.' Our study is the first to systematically assess the impact of psilocybin on timing performance on standardized measures of temporal processing. Results indicate that the serotonin system is selectively involved in duration processing of intervals longer than 2 to 3 seconds and in the voluntary control of the speed of movement. We speculate that psilocybin's selective disruption of longer intervals is likely to be a product of interactions with cognitive dimensions of temporal processing -presumably via 5-HT2A receptor stimulation. PMID- 16714325 TI - Repeated cannabinoid exposure during perinatal, adolescent or early adult ages produces similar longlasting deficits in object recognition and reduced social interaction in rats. AB - There is mounting evidence that chronic cannabis use might result in lasting neurobehavioural changes, although it remains unclear whether vulnerability diminishes with age. The current study compared the effects of cannabinoid exposure at three developmental periods on subsequent measures of memory and anxiety. Male rats aged 4 days (perinatal), 30 days (adolescent) and 56 days (young adult) were injected with vehicle or incremental doses of the cannabinoid receptor agonist CP 55940, daily for 21 consecutive days (0.15, 0.20 or 0.30 mg/kg for 7 days per dose, respectively). Following a 28-day drug-free period, working memory was assessed in an object recognition task. One week later, social anxiety was assessed in a social interaction test. Two days later, generalized anxiety was assessed in an emergence test. Results revealed that CP 55940 impaired working memory and social interaction similarly at all three ages. CP 55940 had no effects in five of six emergence test measures, but a modest but significant reduction in anxiety was noted in one measure following adolescent exposure. We conclude that chronic cannabinoid exposure leads to long-term memory impairments and increased anxiety, irrespective of the age at which drug exposure occurrs. PMID- 16714326 TI - The efficacy of selective serotonin reuptake inhibitors in adult social anxiety disorder: a meta-analysis of double-blind, placebo-controlled trials. AB - Social anxiety disorder is associated with impairment in social and occupational functioning, significant personal distress and a possible economic burden, resulting in a reduction in quality of life. To understand better the efficacy of selective serotonin reuptake inhibitors in social anxiety disorder, randomized, double-blind, placebo-controlled trials were evaluated. Pubmed and PsychINFO electronic databases were searched for social anxiety disorder, social phobia, selective serotonin reuptake inhibitors, citalopram, escitalopram, fluoxetine, fluvoxamine, paroxetine and sertraline. Fifteen published, randomized, double blind, placebo-controlled trials of selective serotonin reuptake inhibitors in social anxiety disorder were identified. Design, subject number, drug and dose, trial length, rating instruments, and baseline and end point data were extracted and then verified independently by a second investigator. Effect sizes were calculated from mean changes in drug and placebo groups in the Liebowitz Social Anxiety Scale and the Sheehan Disability Scale, as well as from other scales where available. For the binary data of the Clinical Global Impression of Change scores, Theta log-odds ratios (the effect-size measure appropriate for binary data) were calculated from proportion changes. Effect sizes for the Liebowitz Social Anxiety Scale ranged from -0.029 to 1.214. Effect sizes for the Sheehan Disability Scale ranged from 0.203 to 0.480 for work, 0.237 to 0.786 for social function, and 0.118 to 0.445 for family function. The Theta log-odds ratios for Clinical Global Impression of Change scores ranged from 0.644 to 3.267. Consistent with previous studies, selective serotonin reuptake inhibitors appear more effective than placebo for social anxiety disorder, with improvement extending into social and occupational function. PMID- 16714327 TI - A case of pancytopenia. AB - Sodium valproate is a commonly used anticonvulsant, particularly in the management of childhood refractory epilepsy. There is a good literature base regarding its haematological effects in this group of patients including the potential for toxic effect on the bone marrow. Valproate is increasingly being used in the treatment of psychiatric conditions, particularly bipolar affective disorder. In this article we describe a case of pancytopenia associated with a valproate level of 166 mg/l. The population of psychiatric patients is different in several ways from the population of children and young adults with epilepsy from whom the existing data comes. The psychiatric patients are older, more likely to misuse substances, more likely to take overdoses and may metabolize valproate more slowly. For these reasons it would be worthwhile investigating the relationship between valproate levels, macrocytosis, platelet counts and B12 levels in this group of patients. The results of such a study would give us a clearer understanding of what the desirable therapeutic range is for valproate in bipolar affective disorder and what, if any, monitoring should be undertaken. PMID- 16714329 TI - Alpha-tocopheryl succinate, in contrast to alpha-tocopherol and alpha-tocopheryl acetate, inhibits prostaglandin E2 production in human lung epithelial cells. AB - The production of prostaglandin E2 (PGE2), a key proinflammatory mediator, is regulated by the availability of its substrate, arachidonic acid (AA), and the activity of the enzyme cyclooxygenase (COX). Increased PGE2 production and COX-2 expression have been observed frequently in specimens from lung cancer patients. Agents that decrease PGE2 production may prevent the initiation and progression of lung cancer. We, therefore, tested the effects of alpha-tocopherol (alphaTOL) analogs on PGE2 production in human lung epithelial cells. Alpha-tocopheryl succinate (alphaTOS), but not alphaTOL or alpha-tocopheryl acetate (alphaTOA), inhibited the phorbol 12-myristate 13-acetate (PMA)-stimulated PGE2 production in three human lung epithelial cell lines (BEAS-2B, H460 and A549 cells). The effect of these compounds on PGE2 production was not correlated with their antioxidant activities, since alphaTOS alone did not inhibit PMA-induced generation of reactive oxygen species. alphaTOS had no effect on PMA-induced AA release or COX 2 expression, although post-incubation with alphaTOS inhibited COX activity and prostaglandin (PGE2 and PGF(2alpha)) production in PMA-stimulated cells. alphaTOS also blocked the COX activity in A549 cells with endogenous high levels of COX enzymes in the absence of PMA stimulation. In addition, the ability of alphaTOS to inhibit COX was affected by AA concentration, suggesting that alphaTOS may compete with AA for interaction with COX proteins. These results suggest that alphaTOS inhibits COX activity, thereby inhibiting PGE2 production in human lung epithelial cells, despite the lack of antioxidant activity. Administration of alphaTOS may block inflammatory responses mediated by PGE2, thereby inhibiting the initiation and progression of lung cancer. PMID- 16714328 TI - Proteinuria, impaired kidney function, and adverse outcomes in people with coronary disease: analysis of a previously conducted randomised trial. AB - OBJECTIVES: To determine whether data on proteinuria are useful for refining estimates of risk based on kidney function alone, and whether the results of kidney function tests can be a useful adjunct to data on proteinuria. DESIGN: Analysis of data from a randomised trial. Impaired kidney function was defined as low glomerular filtration rate (< 60 ml/min/1.73 m2) and proteinuria (> or = 1+ protein) on dipstick urinalysis. SETTING: Study of cholesterol and recurrent events: a randomised trial of pravastatin 40 mg daily versus placebo. PARTICIPANTS: 4098 men and women with previous myocardial infarction. MAIN OUTCOME MEASURES: All cause mortality and cardiovascular events. RESULTS: 371 participants died in nearly 60 months of follow-up. Compared with participants without proteinuria or impaired kidney function, patients with both characteristics were at high risk (hazard ratio 2.39, 95% confidence interval 1.72 to 3.30), and those with only proteinuria or only impaired kidney function were at intermediate risk (1.69, 1.32 to 2.16; 1.41, 1.12 to 1.79, respectively) of dying from any cause. The results were similar for cardiovascular outcomes, including new cases of heart failure, stroke, and coronary death or non-fatal myocardial infarction. A graded increase in the risk of all cause mortality was seen for severity of renal impairment and degree of proteinuria by dipstick. CONCLUSIONS: The presence or absence of proteinuria on dipstick urinalysis may be used to refine estimates of risk based on kidney function alone. PMID- 16714330 TI - Smad-Sp1 complexes mediate TGFbeta-induced early transcription of oncogenic Smad7 in pancreatic cancer cells. AB - The transcription factor Sp1 has been implicated in cell-type-specific activation of transforming growth factor-beta (TGFbeta) target genes in normal epithelial cells as well as in aberrant gene activation by TGFbeta in epithelial tumor cells. Here, we have examined the interaction of Sp1 with components of the Smad signaling cascade and its role in TGFbeta-induced early gene expression in pancreatic cancer cells. Gene expression profiling was carried out in mithramycin A-treated cells to identify Sp1-regulated TGFbeta early response genes. We found that in pancreatic cancer cells Smad proteins and Sp1 cooperatively regulate expression of a distinct set of TGFbeta target genes potentially involved in tumor progression, including MMP-11, cyclin D1 and Smad7. Mechanistically, TGFbeta rapidly induces nuclear translocation of Smad proteins and subsequently stimulates Smad-Sp1 complex formation. Using the Smad7 promoter as a model for Smad-/Sp1-induced early gene activation, we demonstrated that this interaction increases Sp1 binding to GC-rich promoter boxes and results in superinduction of Sp1-mediated transcription. Moreover, inhibition of Sp1-DNA binding or transfection of Sp1-specific siRNA prevents TGFbeta-induced Smad7 expression and consequently enhances Smad signaling in pancreatic cancer cells, as indicated by increased receptor-mediated phosphorylation of Smad3. We thus conclude that Sp1 strongly contributes to the aberrant transcriptional response of transformed epithelial cells to TGFbeta stimulation. PMID- 16714331 TI - Polymorphisms and haplotypes of the NBS1 gene are associated with risk of sporadic breast cancer in non-Hispanic white women C, 8360G>C and 30537G>C) to represent all common (>or=5%) haplotypes reported in the National Institute of Environmental Health Sciences database and to reconstruct haplotypes. In a hospital-based case-control study of 421 non-Hispanic white patients with sporadic breast cancer (C polymorphism. Furthermore, the derived haplotypes were associated with risk in a dose-response manner as the number of variant (risk) alleles (i.e. 8360C, 924C or 30537C) increased (adjusted OR = 1.07, 95% CI = 0.78 1.46 for 1-2 variant alleles; adjusted OR = 2.47, 95% CI = 1.48-4.14 for 3-6 variant alleles; P(trend) = 0.006). These findings suggest that NBS1 polymorphisms and haplotypes may contribute to the etiology of sporadic breast cancer in young non-Hispanic white women. Large studies are warranted to confirm these findings. PMID- 16714332 TI - De novo ICAM-1 synthesis in the mouse lung: model of assessment of protein expression in lungs. AB - Because most studies addressing the regulatory mechanisms of intercellular adhesion molecule (ICAM)-1 expression have used cultured endothelial cells, we set out to develop an isolated mouse lung preparation to study gene and protein expression in its proper cellular context in the organ. Lungs from CD1 mice were isolated and perfused (2 ml/min, 37 degrees C) with a recirculating volume of RPMI 1640 solution supplemented with 3 g/100 ml albumin. Lungs maintained their isogravimetric state for 4 h. Tumor necrosis factor (TNF-alpha; 2,000 U/ml) was added to the perfusate for 0.5, 1, 2, or 3.5 h to induce ICAM-1 expression or lungs received no treatment (control). After quick-freezing the lungs using liquid nitrogen at different time points, the prepared tissue homogenates were analyzed for ICAM-1 protein expression by Western blotting and NF-kappaB activation by electrophoretic mobility shift assay. TNF-alpha caused a progressive increase in NF-kappaB activity after 0.5 h and ICAM-1 protein expression two- to threefold of basal after 2 h. Untreated lungs expressed a low and constant level of ICAM-1 between 0 and 3.5 h. TNF-alpha failed to induce NF kappaB activation and ICAM-1 expression in lungs of NADPH oxidase-deficient mice lacking p47(phox). We disaggregated mouse lungs using collagenase and stained the cells for ICAM-1 and VE-cadherin (used as an endothelial marker) to assess the in situ endothelial-specific expression of ICAM-1. We observed that TNF-alpha challenge resulted in increased ICAM-1 expression in endothelial cells freshly isolated from lungs. These data show the role of NADPH oxidase-derived oxidant signaling in the mechanism of NF-kappaB activation and ICAM-1 expression in mouse lung endothelial cells. Moreover, the general method presented herein has potential value in assessing mechanisms of gene and protein expression in the isolated-perfused mouse lung model. PMID- 16714334 TI - PAR2 activation interrupts E-cadherin adhesion and compromises the airway epithelial barrier: protective effect of beta-agonists. AB - The airway epithelium is an important barrier between the environment and subepithelial tissues. The epithelium is also divided into functionally restricted apical and basolateral domains, and this restriction is dependent on the elements of the barrier. The protease-activated receptor-2 (PAR2) receptor is expressed in airway epithelium, and its activation initiates multiple effects including enhanced airway inflammation and reactivity. We hypothesized that activation of PAR2 would interrupt E-cadherin adhesion and compromise the airway epithelial barrier. The PAR2-activating peptide (PAR2-AP, SLIGRL) caused an immediate approximately 50% decrease in the transepithelial resistance of primary human airway epithelium that persisted for 6-10 min. The decrease in resistance was accompanied by an increase in mannitol flux across the epithelium and occurred in cystic fibrosis transmembrane conductance receptor (CFTR) epithelium pretreated with amiloride to block Na and Cl conductances, confirming that the decrease in resistance represented an increase in paracellular conductance. In parallel experiments, activation of PAR2 interrupted the adhesion of E-cadherin expressing L cells and of primary airway epithelial cells to an immobilized E cadherin extracellular domain, confirming the hypothesis that activation of PAR2 interrupts E-cadherin adhesion. Selective interruption of E-cadherin adhesion with antibody to E-cadherin decreased the transepithelial resistance of primary airway epithelium by >80%. Pretreatment of airway epithelium or the E-cadherin expressing L cells with the long-acting beta-agonist salmeterol prevented PAR2 activation from interrupting E-cadherin adhesion and compromising the airway epithelial barrier. Activation of PAR2 interrupts E-cadherin adhesion and compromises the airway epithelial barrier. PMID- 16714333 TI - Regulation of soluble guanylyl cyclase: looking beyond NO. PMID- 16714335 TI - The epidemic of renal disease--what drives it and what can be done? PMID- 16714336 TI - The role of combination therapy in the management of hypertension. PMID- 16714337 TI - The urinary sediment beyond light microscopical examination. PMID- 16714338 TI - Extracellular secretion of Escherichia coli alkaline phosphatase with a C terminal tag by type I secretion system: purification and biochemical characterization. AB - Type I secretion system (TISS) of Gram-negative bacteria permits proteins to be secreted directly from the cytoplasm to the external medium by a single, energy coupled step. To examine whether this system can be used as an extracellular production system of recombinant proteins, Escherichia coli alkaline phosphatase (AP) was fused to a C-terminal region of Pseudomonas sp. MIS38 lipase (PML) and examined for secretion using the E.coli cells carrying the heterologous TISS. PML is one of the passenger proteins of TISS and contains 12 repetitive sequences and a secretion signal at the C-terminal region. The fusion protein was efficiently secreted to the extracellular medium, while AP was not secreted at all, indicating that the secretion of AP is promoted by a secretion signal of PML. The repetitive sequences were not so important for secretion of the fusion protein, because the secretion level of the fusion protein containing entire repeats ( approximately 10 mg/l culture) was only 2-fold higher than that of the fusion protein without repeats. The fusion protein purified from the culture supernatant existed as a homodimer, like AP, and was indistinguishable from AP in enzymatic properties and stability. PMID- 16714339 TI - A spatial model of the chromatophore vesicles of Rhodobacter sphaeroides and the position of the Cytochrome bc1 complex. AB - The photosynthetic apparatus of purple bacteria is generally considered a well studied and understood system. However, recent atomic force microscopy images of flattened chromatophore vesicles from Rhodobacter sphaeroides restarted a debate about the stoichiometry and positions of the membrane proteins, with the interpretations of the observed images only partly being in agreement with earlier models. The most puzzling observation from the recent images is that the Cytochrome bc(1) complex, which is a central part of the photosynthetic apparatus, seems to be missing on the chromatophore vesicles, even when these were extracted from photosynthetically grown bacteria. From the available information on the geometry of the vesicle and of the proteins we reconstructed here a three-dimensional model vesicle at molecular resolution. Its central feature, also determining its diameter of approximately 45 nm, is an equatorial array of LH1 dimers, lined by a region of LH2 rings. This naturally puts the Cytochrome bc(1) complexes and the ATPase at the vesicle's poles. This spatial model may explain why the vesicle's endcaps with the bc(1) complexes are lost during the preparatory steps of the imaging process together with the ATPase and are therefore absent from the available images. PMID- 16714340 TI - Reconstruction of a kinetic model of the chromatophore vesicles from Rhodobacter sphaeroides. AB - We present a molecular model of a chromatophore vesicle from Rhodobacter sphaeroides. These vesicles are ideal benchmark systems for molecular and systemic simulations, because they have been well studied, they are small, and they are naturally separated from their cellular environment. To set up a photosynthetic chain working under steady-state conditions, we compiled from the experimental literature the specific activities and geometries that have been determined for their constituents. This data then allowed defining the stoichiometries for all membrane proteins. This article contains the kinetic part of the reconstructed model, while the spatial reconstruction is presented in a companion article. By considering the transport properties of the Cytochrome c(2) and ubiquinone pools, we show that their size and oxidation states allow for an efficient buffering of the statistical fluctuations that arise from the small size of the vesicles. Stoichiometric and kinetic considerations indicate that a typical chromatophore vesicle of Rb. sphaeroides with a diameter of 45 nm should contain approximately five bc(1) monomers. PMID- 16714341 TI - Mathematical description of gene regulatory units. AB - Revealing the control mechanisms responsible for the cell's surprisingly well organized functions should lead directly to a better understanding of how the cell adapts to extraordinarily changing environments. A general framework for describing models that can represent diverse biochemical regulatory functions systematically would help not only systematic interpretation of the various models proposed for certain systems but also further understanding of the general control mechanism and design principles underlying different biological systems. This article presents a unified mathematical framework for describing gene regulatory units. The proposed framework is fairly compatible with the classical control theoretical framework, so it should serve as a connecting bridge between engineering control theory and biological control mechanisms. It should also provide a unified view of different regulatory units and facilitate systematic comparison of different mathematical models proposed in a variety of literature. PMID- 16714342 TI - Precise boundary element computation of protein transport properties: Diffusion tensors, specific volume, and hydration. AB - A precise boundary element method for the computation of hydrodynamic properties has been applied to the study of a large suite of 41 soluble proteins ranging from 6.5 to 377 kDa in molecular mass. A hydrodynamic model consisting of a rigid protein excluded volume, obtained from crystallographic coordinates, surrounded by a uniform hydration thickness has been found to yield properties in excellent agreement with experiment. The hydration thickness was determined to be delta = 1.1 +/- 0.1 A. Using this value, standard deviations from experimental measurements are: 2% for the specific volume; 2% for the translational diffusion coefficient, and 6% for the rotational diffusion coefficient. These deviations are comparable to experimental errors in these properties. The precision of the boundary element method allows the unified description of all of these properties with a single hydration parameter, thus far not achieved with other methods. An approximate method for computing transport properties with a statistical precision of 1% or better (compared to 0.1-0.2% for the full computation) is also presented. We have also estimated the total amount of hydration water with a typical -9% deviation from experiment in the case of monomeric proteins. Both the water of hydration and the more precise translational diffusion data hint that some multimeric proteins may not have the same solution structure as that in the crystal because the deviations are systematic and larger than in the monomeric case. On the other hand, the data for monomeric proteins conclusively show that there is no difference in the protein structure going from the crystal into solution. PMID- 16714344 TI - Fiber depolymerization. AB - Depolymerization is, by definition, a crucial process in the reversible assembly of various biopolymers. It may also be an important factor in the pathology of sickle cell disease. If sickle hemoglobin fibers fail to depolymerize fully during passage through the lungs then they will reintroduce aggregates into the systemic circulation and eliminate or shorten the protective delay (nucleation) time for the subsequent growth of fibers. We study how depolymerization depends on the rates of end- and side-depolymerization, k(end) and k(side), which are, respectively, the rates at which fiber length is lost at each end and the rate at which new breaks appear per unit fiber length. We present both an analytic mean field theory and supporting simulations showing that the characteristic fiber depolymerization time tau= square root 1/k(end)k(side) depends on both rates, but not on the fiber length L, in a large intermediate regime 1 << k(side)L(2)/k(end) << (L/d)(2), with d the fiber diameter. We present new experimental data which confirms that both mechanisms are important and shows how the rate of side depolymerization depends strongly on the concentration of CO, acting as a proxy for oxygen. Our theory remains rather general and could be applied to the depolymerization of an entire class of linear aggregates, not just sickle hemoglobin fibers. PMID- 16714343 TI - Flexibility of the neck domain enhances Kinesin-1 motility under load. AB - Kinesin-1 is a dimeric motor protein that moves stepwise along microtubules. A two-stranded alpha-helical coiled-coil formed by the neck domain links the two heads of the molecule, and forces the motor heads to alternate. By exchanging the particularly soft neck region of the conventional kinesin from the fungus Neurospora crassa with an artificial, highly stable coiled-coil we investigated how this domain affects motor kinetics and motility. Under unloaded standard conditions, both motor constructs developed the same gliding velocity. However, in a force-feedback laser trap the mutant showed increasing motility defects with increasing loads, and did not reach wild-type velocities and run lengths. The stall force dropped significantly from 4.1 to 3.0 pN. These results indicate the compliance of kinesin's neck is important to sustain motility under load, and reveal a so far unknown constrain on the imperfect coiled-coil heptad pattern of Kinesin-1. We conclude that coiled-coil structures, a motif encountered in various types of molecular motors, are not merely a clamp for linking two heavy chains to a functional unit but may have specifically evolved to allow motor progression in a viscous, inhomogeneous environment or when several motors attached to a transported vesicle are required to cooperate efficiently. PMID- 16714345 TI - Wavelength-dependent collagen fragmentation during mid-IR laser ablation. AB - Mid-infrared free-electron lasers have proven adept in surgical applications. When tuned to wavelengths between 6 and 7 microm, such lasers remove defined volumes of soft tissue with very little collateral damage. Previous attempts to explain the wavelength-dependence of collateral damage have invoked a wavelength dependent loss of protein structural integrity. However, the molecular nature of this structural failure has been heretofore ill-defined. In this report, we evaluate several candidates for the relevant transition by analyzing the nonvolatile debris ejected during ablation. Porcine corneas were ablated with a free-electron laser tuned to 2.77 or 6.45 microm-wavelengths with matched absorption coefficients for hydrated corneas that respectively target either tissue water or protein. The debris ejected during these ablations was characterized via gel electrophoresis, as well as Fourier transform infrared spectroscopy, micro-Raman and 13C-NMR spectroscopy. We find that high-fluence (240 J/cm2) ablation at 6.45 microm, but not at 2.77 microm, leads to protein fragmentation accompanied by the accumulation of nitrile and alkyne species. The candidate transition most consistent with these observations is scission of the collagen protein backbone at N-alkylamide bonds. Identifying this transition is a key step toward understanding the observed wavelength-dependence of collateral damage in mid-infrared laser ablation. PMID- 16714346 TI - Probing the double layer: effect of image forces on AFM. AB - Force probes such as AFM tips or laser trap latex beads have a dielectric constant much less than that of the water that they displace. Thus when a probe approaches a charged surface under water it will be repelled simply based upon the image forces, and these can be of nN magnitude. PMID- 16714348 TI - Structure of tightly membrane-bound mastoparan-X, a G-protein-activating peptide, determined by solid-state NMR. AB - The structure of mastoparan-X (MP-X), a G-protein activating peptide from wasp venom, in the state tightly bound to anionic phospholipid bilayers was determined by solid-state NMR spectroscopy. Carbon-13 and nitrogen-15 NMR signals of uniformly labeled MP-X were completely assigned by multidimensional intraresidue C-C, N-CalphaCbeta, and N-Calpha-C', and interresidue Calpha-CalphaCbeta, N CalphaCbeta, and N-C'-Calpha correlation experiments. The backbone torsion angles were predicted from the chemical shifts of 13C', 13Calpha, 13Cbeta, and 15N signals with the aid of protein NMR database programs. In addition, two 13C-13C and three 13C-15N distances between backbone nuclei were precisely measured by rotational resonance and REDOR experiments, respectively. The backbone structure of MP-X was determined from the 26 dihedral angle restraints and five distances with an average root-mean-square deviation of 0.6 A. Peptide MP-X in the bilayer bound state formed an amphiphilic alpha-helix for residues Trp3-Leu14 and adopted an extended conformation for Asn2. This membrane-bound conformation is discussed in relation to the peptide's activities to form pores in membranes and to activate G-proteins. This study demonstrates the power of multidimensional solid state NMR of uniformly isotope-labeled molecules and distance measurements for determining the structures of peptides bound to lipid membranes. PMID- 16714347 TI - Modeling the role of incisures in vertebrate phototransduction. AB - Phototransduction is mediated by a G-protein-coupled receptor-mediated cascade, activated by light and localized to rod outer segment (ROS) disk membranes, which, in turn, drives a diffusion process of the second messengers cGMP and Ca2+ in the ROS cytosol. This process is hindered by disks-which, however, bear physical cracks, known as incisures, believed to favor the longitudinal diffusion of cGMP and Ca2+. This article is aimed at highlighting the biophysical functional role and significance of incisures, and their effect on the local and global response of the photocurrent. Previous work on this topic regarded the ROS as well stirred in the radial variables, lumped the diffusion mechanism on the longitudinal axis of the ROS, and replaced the cytosolic diffusion coefficients by effective ones, accounting for incisures through their total patent area only. The fully spatially resolved model recently published by our group is a natural tool to take into account other significant details of incisures, including their geometry and distribution. Using mathematical theories of homogenization and concentrated capacity, it is shown here that the complex diffusion process undergone by the second messengers cGMP and Ca2+ in the ROS bearing incisures can be modeled by a family of two-dimensional diffusion processes on the ROS cross sections, glued together by other two-dimensional diffusion processes, accounting for diffusion in the ROS outer shell and in the bladelike regions comprised by the stack of incisures. Based on this mathematical model, a code has been written, capable of incorporating an arbitrary number of incisures and activation sites, with any given arbitrary distribution within the ROS. The code is aimed at being an operational tool to perform numerical experiments of phototransduction, in rods with incisures of different geometry and structure, under a wide spectrum of operating conditions. The simulation results show that incisures have a dual biophysical function. On the one hand, since incisures line up from disk to disk, they create vertical cytoplasmic channels crossing the disks, thus facilitating diffusion of second messengers; on the other hand, at least in those species bearing multiple incisures, they divide the disks into lobes like the petals of a flower, thus confining the diffusion of activated phosphodiesterase and localizing the photon response. Accordingly, not only the total area of incisures, but their geometrical shape and distribution as well, significantly influence the global photoresponse. PMID- 16714349 TI - Role of solvent on protein-matrix coupling in MbCO embedded in water-saccharide systems: a Fourier transform infrared spectroscopy study. AB - Embedding protein in sugar systems of low water content enables one to investigate the protein dynamic-structure function in matrixes whose rigidity is modulated by varying the content of residual water. Accordingly, studying the dynamics and structure thermal evolution of a protein in sugar systems of different hydration constitutes a tool for disentangling solvent rigidity from temperature effects. Furthermore, studies performed using different sugars may give information on how the detailed composition of the surrounding solvent affects the internal protein dynamics and structural evolution. In this work, we compare Fourier transform infrared spectroscopy measurements (300-20 K) on MbCO embedded in trehalose, sucrose, maltose, raffinose, and glucose matrixes of different water content. At all the water contents investigated, the protein solvent coupling was tighter in trehalose than in the other sugars, thus suggesting a molecular basis for the trehalose peculiarity. These results are in line with the observation that protein-matrix phase separation takes place in lysozyme-lactose, whereas it is absent in lysozyme-trehalose systems; indeed, these behaviors may respectively be due to the lack or presence of suitable water mediated hydrogen-bond networks, which match the protein surface to the surroundings. The above processes might be at the basis of pattern recognition in crowded living systems; indeed, hydration shells structural and dynamic matching is first needed for successful come together of interacting biomolecules. PMID- 16714350 TI - Preferential hydration of DNA: the magnitude and distance dependence of alcohol and polyol interactions. AB - The physical forces that underlie the exclusion of solutes from macromolecular surfaces can be probed in a similar way as the measurement of forces between macromolecules in condensed arrays using the osmotic stress technique and x-ray scattering. We report here the dependence of alcohol exclusion or, equivalently, the preferential hydration of DNA on the spacing between helices in condensed arrays. The actual forces describing exclusion are quite different from the commonly assumed steric crowding coupled with weak binding. For a set of 12 nonpolar alcohols, exclusion is due to repulsive hydration interactions with the charged DNA surface. Exclusion amplitudes do not depend simply on size, but rather on the balance between alkyl carbons and hydroxyl oxygens. Polyols are included at very close spacings. The distance dependence of polyol inclusion, however, is quite different from nonpolar alcohol exclusion, suggesting the underlying mechanism of interaction is different. PMID- 16714351 TI - Molecular dynamics simulations of NAD+-induced domain closure in horse liver alcohol dehydrogenase. AB - Horse liver alcohol dehydrogenase is a homodimer, the protomer having a coenzyme binding domain and a catalytic domain. Using all available x-ray structures and 50 ns of molecular dynamics simulations, we investigated the mechanism of NAD+ induced domain closure. When the well-known loop at the domain interface was modeled to its conformation in the closed structure, the NAD+-induced domain closure from the open structure could be simulated with remarkable accuracy. Native interactions in the closed structure between Arg369, Arg47, His51, Ala317, Phe319, and NAD+ were seen to form at different stages during domain closure. Removal of the Arg369 side-chain charge resulted in the loss of the tendency to close, verifying that specific interactions do help drive the domains closed. Further simulations and a careful analysis of x-ray structures suggest that the loop prevents domain closure in the absence of NAD+, and a cooperative mechanism operates between the subunits for domain closure. This cooperative mechanism explains the role of the loop as a block to closure because in the absence of NAD+ it would prevent the occurrence of an unliganded closed subunit when the other subunit closes on NAD+. Simulations that started with one subunit open and one closed supported this. PMID- 16714352 TI - Quantifying the magnetic advantage in magnetotaxis. AB - Magnetotactic bacteria are characterized by the production of magnetosomes, nanoscale particles of lipid bilayer encapsulated magnetite, that act to orient the bacteria in magnetic fields. These magnetosomes allow magneto-aerotaxis, which is the motion of the bacteria along a magnetic field and toward preferred concentrations of oxygen. Magneto-aerotaxis has been shown to direct the motion of these bacteria downward toward sediments and microaerobic environments favorable for growth. Herein, we compare the magneto-aerotaxis of wild-type, magnetic Magnetospirillum magneticum AMB-1 with a nonmagnetic mutant we have engineered. Using an applied magnetic field and an advancing oxygen gradient, we have quantified the magnetic advantage in magneto-aerotaxis as a more rapid migration to preferred oxygen levels. Magnetic, wild-type cells swimming in an applied magnetic field more quickly migrate away from the advancing oxygen than either wild-type cells in a zero field or the nonmagnetic cells in any field. We find that the responses of the magnetic and mutant strains are well described by a relatively simple analytical model, an analysis of which indicates that the key benefit of magnetotaxis is an enhancement of a bacterium's ability to detect oxygen, not an increase in its average speed moving away from high oxygen concentrations. PMID- 16714354 TI - Distributed model of peritoneal fluid absorption. AB - The process of water reabsorption from the peritoneal cavity into the surrounding tissue substantially decreases the net ultrafiltration in patients on peritoneal dialysis. The goal of this study was to propose a mathematical model based on data from clinical studies and animal experiments to describe the changes in absorption rate, interstitial hydrostatic pressure, and tissue hydration caused by increased intraperitoneal pressure after the initiation of peritoneal dialysis. The model describes water transport through a deformable, porous tissue after infusion of isotonic solution into the peritoneal cavity. Blood capillary and lymphatic vessels are assumed to be uniformly distributed within the tissue. Starling's law is applied for a description of fluid transport through the capillary wall, and the transport within the interstitium is modeled by Darcy's law. Transport parameters such as interstitial fluid volume ratio, tissue hydraulic conductance, and lymphatic absorption in the tissue are dependent on local interstitial pressure. Numerical simulations show the strong dependence of fluid absorption and tissue hydration on the values of intraperitoneal pressure. Our results predict that in the steady state only approximately 20-40% of the fluid that flows into the tissue from the peritoneal cavity is absorbed by the lymphatics situated in the tissue, whereas the larger (60-80%) part of the fluid is absorbed by the blood capillaries. PMID- 16714353 TI - Membrane lateral diffusion and capture of CFTR within transient confinement zones. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) channel interacts with scaffolding and other proteins that are expected to restrict its lateral movement, yet previous studies have reported predominantly free diffusion. We examined the lateral mobility of CFTR channels on live baby hamster kidney cells using three complementary methods. Channels bearing an extracellular biotinylation target sequence were labeled with streptavidin conjugated with fluorescent dyes (Alexa Fluor 488 or 568) or quantum dots (qDot605). Fluorescence recovery after photobleaching and image correlation spectroscopy of the dye labeled channels revealed a significant immobile population ( approximately 50%), which was confirmed by direct single particle tracking (SPT) of qDot605-labeled CFTR. Adding 10 histidine residues at the C-terminus of CFTR to mask the postsynaptic density 95, Discs large, ZO-1 (PDZ) binding motif abolished its association with EBP50/NHERF1, reduced the immobile fraction, and increased mobility. Other interactions that are not normally detected on this timescale became apparent when binding of PDZ domain proteins was disrupted. SPT revealed that CFTR(His-10) channels diffuse randomly, become immobilized for periods lasting up to 1 min, and in some instances are recaptured at the same location. The impact of transient confinement on the measured diffusion using the three fluorescence techniques were assessed using computer simulations of the biological experiments. Finally, the impact of endosomal CFTR on mobility measurements was assessed by fluorescence correlation spectroscopy. These results reveal unexpected features of CFTR dynamics which may influence its ion channel activity. PMID- 16714355 TI - Signaling and regulation of endothelial cell survival by angiopoietin-2. AB - Angiopoietins are ligands for endothelial cell-specific Tie-2 receptors. Whereas angiopoietin-1 (Ang-1) activates these receptors and promotes cell survival, migration, and sprouting, little information is available regarding how Ang-2 influences these cells. In this study, we evaluated signaling pathways and biological effects of physiological concentrations of Ang-2 in cultured human umbilical vein endothelial cells. Ang-2 at 150 and 300 ng/ml elicited a transient (reaching peak values within 15 min of exposure) increase in the phosphorylation of Tie-2 receptors, protein kinase B (Akt), ERK1/2, and p38 members of the mitogen-activated protein kinases. However, unlike Ang-1, Ang-2 significantly inhibited JNK/SAPK phosphorylation. When vascular endothelial growth factor (VEGF) was present along with Ang-2, ERK1/2 phosphorylation was inhibited, whereas augmentation of Ang-1-induced ERK1/2 phosphorylation was triggered by VEGF. Ang-2 treatment had no effect on cell migration and in vitro wound healing but significantly attenuated serum deprivation-induced apoptosis and promoted survival. These effects were completely reversed by phosphatidylinositol 3 (PI3) kinase and ERK1/2 inhibitors but were augmented by an inhibitor of the p38 pathway. These results suggest that Ang-2 promotes endothelial cell survival through the ERK1/2 and PI3-kinase pathways and that this angiopoietin is not a strong promoter of endothelial cell migration. We also conclude that the nature of interactions in terms of ERK1/2 activation between Ang-2 and VEGF is different from that of Ang-1 and VEGF. PMID- 16714356 TI - Progression of coronary and mesenteric vascular dysfunction in Zucker obese and Zucker diabetic fatty rats. AB - We investigated the progression of vascular dysfunction associated with the metabolic syndrome with and without hyperglycemia in lean, Zucker obese, and Zucker diabetic fatty (ZDF) rats. Responses of aorta and small coronary and mesenteric arteries were measured to endothelium-dependent and -independent vasodilators. Indices of oxidative stress were increased in serum from ZDF rats throughout the study, whereas values were increased in Zucker obese rats later in the study [thiobarbituric acid reactive substances: 0.45 +/- 0.02, 0.59 +/- 0.03 (P < 0.05), and 0.58 +/- 0.03 (P < 0.05) mug/ml in serum from 28- to 40-wk-old lean, Zucker obese, and ZDF rats, respectively]. Acetylcholine (ACh)-induced relaxation was not altered in vessels from lean animals from 8-40 wk. ACh-induced relaxation was nearly abolished in coronary arteries from 28- to 36-wk-old Zucker obese rats and by 16-36 wk in ZDF rats and was attenuated in aorta and mesenteric vessels from ZDF rats [%relaxation to 10 muM ACh: 72.2 +/- 7.1, 17.9 +/- 5.9 (P < 0.05), and 23.0 +/- 4.5 (P < 0.05) in coronary vessels; and 67.9 +/- 9.2, 50.1 +/ 5.5, and 42.3 +/- 4.7 (P < 0.05) in mesenteric vessels from 28- to 40-wk-old lean, Zucker obese, and ZDF rats, respectively]. The attenuated ACh-induced relaxation was improved when vessels were incubated with tiron, suggesting superoxide as a mechanism of endothelial dysfunction. Sodium nitroprusside induced relaxation was not altered in aorta or coronary arteries and was potentiated in mesenteric arteries from Zucker obese rats. Our data suggest that diabetes enhances the progression of vascular dysfunction. Increases in indices of oxidative stress precede the development of dysfunction and may serve as a marker of endothelial damage. PMID- 16714357 TI - Cardioprotection initiated by reactive oxygen species is dependent on activation of PKCepsilon. AB - To examine whether cardioprotection initiated by reactive oxygen species (ROS) is dependent on protein kinase Cepsilon (PKCepsilon), isolated buffer-perfused mouse hearts were randomized to four groups: 1) antimycin A (AA) (0.1 microg/ml) for 3 min followed by 10 min washout and then 30 min global ischemia (I) and 2 h reperfusion (R); 2) controls of I/R alone; 3) AA bracketed with 13 min of N-2 mercaptopropionyl- glycine (MPG) followed by I/R; and 4) MPG (200 microM) alone, followed by I/R. Isolated adult rat ventricular myocytes (ARVM) were exposed to AA (0.1 microg/ml), and lucigenin was used to measure ROS production. Murine hearts and ARVM were exposed to AA (0.1 microg/ml) with or without MPG, and PKCepsilon translocation was measured by cell fractionation and subsequent Western blot analysis. Finally, the dependence of AA protection on PKCepsilon was determined by the use of knockout mice (-/-) lacking PKCepsilon. AA exposure caused ROS production, which was abolished by the mitochondrial uncoupler mesoxalonitrile 4-trifluoromethoxyphenylhydrazone. In addition, AA significantly reduced the percent infarction-left ventricular volume compared with control I/R (26 +/- 4 vs. 43 +/- 2%; P < 0.05). Bracketing AA with MPG caused a loss of protection (52 +/- 7 vs. 26 +/- 4%; P < 0.05). AA caused PKCepsilon translocation only in the absence of MPG, and protection was lost on the pkcepsilon(-/-) background (38 +/- 3 vs. 15 +/- 4%; P < 0.001). AA causes ROS production, on which protection and PKCepsilon translocation depend. In addition, protection is absent in PKCepsilon null hearts. Our results imply that, in common with ischemic preconditioning, PKCepsilon is crucial to ROS-mediated protection. PMID- 16714358 TI - Spatial frequency content in optical mapping of cardiac cell monolayers. PMID- 16714359 TI - Angiotensin II-mediated oxidative stress and procollagen-1 expression in cardiac fibroblasts: blockade by pravastatin and pioglitazone. AB - Angiotensin II (ANG II), a product of renin-angiotensin system activation, enhances collagen synthesis, which is a key event in cardiac remodeling after myocardial infarction. Inhibition of cardiac remodeling is now a target of multiple therapies, including 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, commonly known as statins, and peroxisome proliferator-activated receptor-gamma (PPAR-gamma) ligands. We examined the potential antifibrotic effect of the combination of a statin (pravastatin) and a PPAR-gamma ligand (pioglitazone) in ANG II-treated mouse cardiac fibroblasts. ANG II treatment induced procollagen-1 expression, which was inhibited by pravastatin and pioglitazone in a dose-dependent fashion. Pretreatment of fibroblasts with low therapeutic concentrations of either pravastatin (0.1 microM) or pioglitazone (5 microM) only slightly decreased ANG II-induced NADPH oxidase expression, superoxide anion production, and procollagen-1 expression; however, the combination of pravastatin and pioglitazone markedly modulated these effects of ANG II. The combination also blocked ANG II-mediated p38 MAPK and p44/42 MAPK activation. Electrophoretic mobility shift assay showed that ANG II activated transcription factors NF-kappaB and activator protein-1 (AP-1). Although pravastatin and pioglitazone alone had a variable effect on NF-kappaB and AP-1 activation, their combination exerted a potent inhibitory effect on the activation of both NF-kappaB and AP-1. The effects of pravastatin and pioglitazone in combination on superoxide generation and procollagen-1 expression mimicked those of alpha-tocopherol and gamma-tocopherol, two potent antioxidants. Thus it appears that there is a positive interaction between pravastatin and pioglitazone in modulating ANG II-mediated oxidative stress, inhibiting MAPK activation, and procollagen-1 expression. PMID- 16714360 TI - Compensatory growth of coronary arterioles in postinfarcted heart: regional differences in DNA synthesis and growth factor/receptor expression patterns. AB - Previous studies have not addressed regional differences in adaptive arteriolar growth in the surviving left ventricular (LV) myocardium after infarction in appropriately aged animals, namely middle-aged or older. Accordingly, we examined the adaptive postinfarction growth of arterioles in two distinct regions, i.e., the LV free wall (LVFW) and septum, of middle-aged rats. We induced a myocardial infarction (MI) in 12-mo-old rats to analyze 1) protein expression in VEGF/Flt 1/Flk-1 and angiopoietin (Ang)-1/Ang-2/Tie-2 systems, 2) the arteriolar DNA synthesis, 3) the extent of the arteriolar bed, and 4) the alteration in minimal coronary vascular resistance. In both regions, arteriolar DNA synthesis was activated between days 4 and 7 after MI. Whereas in the LVFW the degree of DNA synthesis declined between days 11 and 14 post-MI, it continued to rise in the septum, and at day 14, the percentage of the arterioles undergoing DNA synthesis was comparable in the LVFW and the septum (9.7 +/- 1.6 and 7 +/- 2.1%, respectively). Arteriolar DNA synthesis was mainly associated with upregulation of Ang-2 and Tie-2 in both LV regions. Although 4 wk after MI the arteriolar beds in the LVFW and the septum expanded to the size of sham-operated rats, this growth did not compensate for the greater minimal coronary vascular resistance in the former. Thus our findings suggest that 1) the dynamics in adaptive arteriolar growth were similar between the two regions, despite a delay in the septum; and 2) the perfusion deficit in post-MI rats cannot be accounted for by inadequate adaptive growth of arterioles. PMID- 16714361 TI - Leg crossing improves orthostatic tolerance in healthy subjects: a placebo controlled crossover study. AB - Vasovagal syncope is the most common cause of transient loss of consciousness, and recurrent vasovagal fainting has a profound impact on quality of life. Physical countermaneuvers are applied as a means of tertiary prevention but have so far only proven useful at the onset of a faint. This placebo-controlled crossover study tested the hypothesis that leg crossing increases orthostatic tolerance. Nine naive healthy subjects [6 females, median age 25 yr (range 20-41 yr), mean body mass index 23 (SD 2)] were subjected to passive head-up tilt combined with a graded lower body negative pressure challenge (20, 40, and 60 mmHg) determining orthostatic tolerance thrice, in randomized order: 1) control, 2) with leg crossing, and 3) with oral placebo. Blood pressure (Finometer), heart rate, and changes in thoracic blood volume (impedance), stroke volume, and cardiac output (Modelflow) were followed during orthostatic stress. Primary outcome was time to presyncope (systolic blood pressure /=140 beats/min). With leg crossing, orthostatic tolerance increased from 26 +/- 2 to 34 +/- 2 min (placebo 23 +/- 3 min, P < 0.001). During leg crossing, mean arterial pressure (81 vs. 81 mmHg) and cardiac output (95 vs. 94% supine) remained unchanged; heart rate increase was lower (13 vs. 18 beats/min, P < 0.05); stroke volume was higher (79 vs. 74% supine, P < 0.05); and there was a trend toward lower thoracic impedance. Leg crossing increases orthostatic tolerance in healthy human subjects. As a measure of prevention, it is a worthwhile addition to the management of vasovagal syncope. PMID- 16714362 TI - Allometric scaling of wall shear stress from mice to humans: quantification using cine phase-contrast MRI and computational fluid dynamics. AB - Allometric scaling laws relate structure or function between species of vastly different sizes. They have rarely been derived for hemodynamic parameters known to affect the cardiovascular system, e.g., wall shear stress (WSS). This work describes noninvasive methods to quantify and determine a scaling law for WSS. Geometry and blood flow velocities in the infrarenal aorta of mice and rats under isoflurane anesthesia were quantified using two-dimensional magnetic resonance angiography and phase-contrast magnetic resonance imaging at 4.7 tesla. Three dimensional models constructed from anatomic data were discretized and used for computational fluid dynamic simulations using phase-contrast velocity imaging data as inlet boundary conditions. WSS was calculated along the infrarenal aorta and compared between species to formulate an allometric equation for WSS. Mean WSS along the infrarenal aorta was significantly greater in mice and rats compared with humans (87.6, 70.5, and 4.8 dyn/cm(2), P < 0.01), and a scaling exponent of -0.38 (R(2) = 0.92) was determined. Manipulation of the murine genome has made small animal models standard surrogates for better understanding the healthy and diseased human cardiovascular system. It has therefore become increasingly important to understand how results scale from mouse to human. This noninvasive methodology provides the opportunity to serially quantify changes in WSS during disease progression and/or therapeutic intervention. PMID- 16714363 TI - Activated neutrophil-mediated sickle red blood cell adhesion to lung vascular endothelium: role of phosphatidylserine-exposed sickle red blood cells. AB - Activated neutrophils (ANs) increase sickle red blood cell (SRBC) retention/adhesion in the pulmonary circulation. This study investigates the role of neutrophil activation and SRBC retention/adhesion in the pulmonary circulation through a mechanism that involves increasing phosphatidylserine (PS) exposure on the external membrane surface of the SRBCs (PS-exposed). With the use of flow cytometry, double-labeling studies were performed with a calcium-dependent phospholipid-binding protein, annexin V-fluorescein isothiocyanate fluorescence, and the erythroid-specific marker glycophorin A to assess for the percentage of PS-exposed normal and SRBCs at baseline and after coincubation with ANs. Additional studies were performed that assessed retention/adhesion of SRBCs in the isolated rat lung using (51)Cr-labeled SRBC alone, SRBC + AN, SRBC + AN + zileuton, and SRBC + AN + annexin V. Specific activities of lung and perfusate were measured, and the number of retained SRBCs per gram lung was calculated. Flow cytometry demonstrated that ANs increased the percentage of PS-exposed normal and SRBCs. The 5-lipoxygenase inhibitor zileuton attenuated AN-mediated increases in PS-exposed SRBCs and decreased SRBC retention/adherence in the lung on histological sections. Similarly, in the isolated perfused lung and in histological lung sections, retention/adherence of SRBCs cloaked with annexin V was attenuated in the presence of ANs. We conclude that ANs enhance the adhesion of SRBCs to vascular endothelium by increasing red blood cell membrane externalization of PS. Zileuton attenuation of AN-mediated SRBC PS externalization suggests that a 5-lipoxygenase product(s), secreted by the AN, plays a vital role in altering the adhesive properties of PS-exposed SRBCs to vascular endothelium. PMID- 16714364 TI - Calculation of threshold and saturation points of sigmoidal baroreflex function curves. AB - The logistic sigmoid function curve provides an accurate description of the baroreflex input-output relationship and is the most commonly used equation for this purpose. The threshold (Thr) and saturation (Sat) values for the baroreflex are commonly defined as the values of mean arterial pressure (MAP) at which the reflexly controlled variable (e.g., heart rate or sympathetic nerve activity) is within 5% of the upper or lower plateau, respectively, of the sigmoid function. These values are referred to here as Thr(5%) and Sat(5%). In many studies, Thr and Sat are calculated with the equations Thr = A(3) - 2.0/A(2) and Sat = A(3) + 2.0/A(2), where A(3) is the value of MAP at the point where the reflexly controlled variable is at the midpoint of its range and A(2) is the gain coefficient. Although it is commonly stated that the values of Thr and Sat calculated with these equations represent Thr(5%) and Sat(5%), we show here that instead they are significantly greater and less than Thr(5%) and Sat(5%), respectively. Furthermore, the operating range (difference between Thr and Sat) calculated with these equations is 32% less than the difference between Thr(5%) and Sat(5%). We further show that the equations that provide correct values of Thr(5%) and Sat(5%) are Thr(5%) = A(3) - 2.944/A(2) and Sat(5%) = A(3) + 2.944/A(2). We propose that these be used as the standard equations for calculating threshold and saturation values when a logistic sigmoid function is used to model the open-loop baroreflex function curve. PMID- 16714365 TI - Wave reflection effects in the central circulation of American alligators (Alligator mississippiensis): what the heart sees. AB - A large central compliance is thought to dominate the hemodynamics of all vertebrates except birds and mammals. Yet large crocodilians may adumbrate the avian and mammalian condition and set the stage for significant wave transmission (reflection) effects, with potentially detrimental impacts on cardiac performance. To investigate whether crocodilians exhibit wave reflection effects, pressures and flows were recorded from the right aorta, carotid artery, and femoral artery of six adult, anesthetized American alligators (Alligator mississippiensis) during control conditions and after experimentally induced vasodilation and constriction. Hallmarks of wave reflection phenomena were observed, including marked differences between the measured profiles for flow and pressure, peaking of the femoral pressure pulse, and a diastolic wave in the right aortic pressure profile. Pulse wave velocity and peripheral input impedance increased with progressive constriction, and thus changes in both the timing and magnitude of reflections accounted for the altered reflection effects. Resolution of pressure and flow waves into incident and reflected components showed substantial reflection effects within the right aorta, with reflection coefficients at the first harmonic approaching 0.3 when constricted. Material properties measured from isolated segments of blood vessels revealed a major reflection site at the periphery and, surprisingly, at the junction of the truncus and right aorta. Thus, while our results clearly show that significant wave reflection phenomena are not restricted to birds and mammals, they also suggest that rather than cope with potential negative impacts of reflections, the crocodilian heart simply avoids them because of a large impedance mismatch at the truncus. PMID- 16714366 TI - Feasibility, acceptability, and quality of Internet-administered adolescent health promotion in a preventive-care setting. AB - An Internet tool for monitoring, personalized feedback and referral was developed to support routine adolescent preventive care and was compared with usual practice using paper and pencil (P&P). A total of 1071 students (average age 15 years) from seven secondary schools were randomly assigned to the Internet or P&P group. The Internet group received a health and health-behavior assessment, tailored feedback on health and health behavior (specifically fruit consumption), and an online referral to see a physician/nurse if necessary. The P&P group received the same assessment, preprinted generic advice on fruit consumption and a mailed referral (where applicable). Students and physicians/nurses completed evaluation forms to assess indicators of feasibility, acceptability (i.e. satisfaction) and quality of each administration mode. Student participation rate was 87%. The electronic health feedback was positively evaluated. Students perceived the Internet-tailored fruit advice as more pleasant, more personally targeted and more enjoyable, but less credible than the generic preprinted advice (P < 0.01). No differences in indicators of acceptability and quality of consultation were found (P > or = 0.05). Thus, the Internet can be a valuable tool to support physicians/nurses in the field of preventive care. It is recommended to further optimize and evaluate the Internet as a tool. PMID- 16714367 TI - Taking part counts: adolescents' experiences of the transition from inactivity to active participation in school-based physical education. AB - Identifying ways to increase and sustain active living among young people represents a priority for health promotion interventions. This qualitative study explored the experiences of adolescent boys and girls in the United Kingdom (aged 14-15 years) who had made the transition from inactivity to active participation in physical education (PE). The setting was a secondary school that had modified the PE programme, with the aim of increasing participation rates. Thirty-one, self-identified, formerly 'PE adverse' students were interviewed in focus groups in the school setting. Previously, respondents felt that they had been constructed as physically marginal individuals. Having a physically 'marginal body' was perceived as being detrimental to emotional health. The new PE resulted in respondents possessing a performing and achieving physical identity. Boys gained a sense of security in their physical identity and cultural change had removed the aggression from interacting with other physically active peers. Girls spoke of increased self-confidence, including acquiring the psychological resources to participate in community-based activities. Effective health promotion interventions for inactive adolescents are likely to move beyond solely endorsing the benefits of physical activity or increasing choice and instead address the potential emotional risks and value of participation from the adolescents' perspective. PMID- 16714368 TI - Molecular analysis of the IVS8-T splice variant 5T and M470V exon 10 missense polymorphism in Iranian males with congenital bilateral absence of the vas deferens. AB - Congenital bilateral absence of the vas deferens (CBAVD) is responsible for 2-6% of male infertility in which mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene have been identified. To investigate CBAVD at the molecular level in Iran, we have characterized the mutations in the CFTR gene in 106 patients with this condition. None had clinical manifestations of cystic fibrosis (CF). We also analysed a DNA variant (the 5T allele) in a noncoding region of CFTR, which causes reduced levels of the normal CFTR protein and M470V exon 10 missense polymorphism. Five of the 106 patients with CBAVD had mutations in both copies of the CFTR gene, and none of them had the 5T allele. Eighty-five patients had a mutation in at least one copy of CFTR, and of these patients, 46 had one 5T allele (in 11 cases, two alleles and in 35 cases, just one allele of 5T was detected). In 21 patients, no CFTR and 5T mutations were found (19.81%). 5T/M470 genotype was found in 19 patients, 5T/V470 was found in 3 and 5T with heterozygote form of M470V was found in 24 CBAVD patients. In CBAVD patients, 28 F508del carriers were identified. Most of our patients with CBAVD have mutations in the CFTR gene. The combination of the 5T allele in one copy of the CFTR gene with a CF mutation in the other copy is the most common cause of CBAVD in Iran. The 5T allele mutation has a wide range of clinical presentations and revealed a high frequency, occurring in patients with CBAVD or moderate forms of CF and infertile men. PMID- 16714369 TI - Novel function of ovarian growth factors: combined studies by DNA microarray, biochemical and physiological approaches. AB - Owing to the development of the DNA microarray technique, modulation of gene function can be studied systematically. Considerable attention has been focused on members of the growth factor family to elucidate the main regulators of oocyte maturation and ovarian follicle rupture. Among these growth factors, it was found both in rodents and in humans that amphiregulin (Ar) and epiregulin (Ep) of the epidermal growth factor (EGF) family were dramatically up-regulated by gonadotrophins in the intact ovary and in primary granulosa cells, respectively. Their role in cumulus expansion and oocyte maturation was established in rodents, and their formation under LH stimulation in granulosa cells was demonstrated in humans. To be activated, Ar and Ep must be cleaved by A Disintegrin And Metalloproteinases (ADAMs) family. However, the precise processing of Ar and Ep by the cumulus cells is still obscure. Future investigations using DNA microarray technique may reveal the repertoire of genes activated in Ar- and Ep-stimulated cumulus cells and may help elucidate the molecular basis of ovulation. PMID- 16714370 TI - Kinetic isotope effects implicate a single oxidant for cytochrome P450-mediated O dealkylation, N-oxygenation, and aromatic hydroxylation of 6-methoxyquinoline. AB - One major point of controversy in the area of cytochrome P450 (P450)-mediated oxidation reactions is the nature of the active-oxygen species. A number of hypotheses have been advanced which implicate a second oxidant besides the iron oxo species designated as compound I (Cpd 1). This oxygen is thought to be either an iron-hydroperoxy species (Cpd 0) or a second spin-state of Cpd 1. Very little information is available on what fraction of P450 oxidations is mediated by the two different oxidants. Herein, we report results on three cytochrome P450 mediated reactions: O-dealkylation, N-oxygenation, and aromatic hydroxylation, which occur by three distinct chemical mechanisms. We have used kinetic isotope effects to test for branching from O-demethylation to N-oxygenation and aromatic hydroxylation, using 6-methoxyquinoline and 2H3-6-methoxyquinoline as substrates for P4501A2. Identical large inverse isotope effects on Vmax/Km are obtained for the formation of both the N-oxide and the phenol. This indicates that all three reactions occur through the same enzyme-substrate complex and, thus, through a single iron-oxygen species. The nature of the iron-oxygen species is less certain but is more likely to be iron-oxo Cpd 1, given the energetics of these reactions. PMID- 16714371 TI - Induction of CYP1A and cyp2-mediated arachidonic acid epoxygenation and suppression of 20-hydroxyeicosatetraenoic acid by imidazole derivatives including the aromatase inhibitor vorozole. AB - Cytochrome P450 (P450) enzymes metabolize the membrane lipid arachidonic acid to stable biologically active epoxides [eicosatrienoic acids (EETs)] and 20 hydroxyeicosatetraenoic acid (20-HETE). These products have cardiovascular activity, primarily acting as vasodilators and vasoconstrictors, respectively. EET formation can be increased by the prototype CYP1A or CYP2 inducers, 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) or phenobarbital (PB), respectively. We report here that imidazole derivative drugs: the anthelminthics, albendazole and thiabendazole; the proton pump inhibitor, omeprazole; the thromboxane synthase inhibitor, benzylimidazole; and the aromatase (CYP19) inhibitor vorozole (R76713, racemate; and R83842, (+) enantiomer) increased hepatic microsomal EET formation in a chick embryo model. Albendazole increased EETs by transcriptional induction of CYP1A5 and the others by combined induction of CYP1A5 and CYP2H, the avian orthologs of mammalian CYP1A2 and CYP2B, respectively. All inducers increased formation of the four EET regioisomers, but TCDD and albendazole had preference for 5,6-EET and PB and omeprazole for 14,15-EET. Vorozole, benzylimidazole, and TCDD also suppressed 20-HETE formation. Vorozole was a remarkably effective and potent inducer of multiple hepatic P450s at a dose range which overlapped its inhibition of ovarian aromatase. Increased CYP1A activity in mouse Hepa 1-6 and human HepG2 cells by vorozole and other imidazole derivatives demonstrated applicability of the findings to mammalian cells. The findings suggest that changes in P450-dependent arachidonic acid metabolism may be a new source of side effects for drugs that induce CYP1A or CYP2. They demonstrate further that in vivo induction of multiple hepatic P450s produces additive increases in arachidonic acid epoxygenase activity and can occur concurrently with inhibition of ovarian aromatase activity. PMID- 16714372 TI - The environmental pollutant and carcinogen 3-nitrobenzanthrone and its human metabolite 3-aminobenzanthrone are potent inducers of rat hepatic cytochromes P450 1A1 and -1A2 and NAD(P)H:quinone oxidoreductase. AB - 3-Nitrobenzanthrone (3-NBA), a suspected human carcinogen occurring in diesel exhaust and air pollution, and its human metabolite 3-aminobenzanthrone (3-ABA) were investigated for their ability to induce biotransformation enzymes in rat liver and the influence of such induction on DNA adduct formation by the compounds. Rats were treated (i.p.) with 0.4, 4, or 40 mg/kg body weight 3-NBA or 3-ABA. When hepatic cytosolic fractions from rats treated with 40 mg/kg body weight 3-NBA or 3-ABA were incubated with 3-NBA, DNA adduct formation, measured by 32P-postlabeling analysis, was 10-fold higher in incubations with cytosols from pretreated rats than with controls. The increase in 3-NBA-derived DNA adduct formation corresponded to a dose-dependent increase in protein levels and enzymatic activity of NAD(P)H:quinone oxidoreductase (NQO1). NQO1 is the major enzyme reducing 3-NBA in human and rat livers. Incubations of 3-ABA with hepatic microsomes of rats treated with 3-NBA or 3-ABA (40 mg/kg body weight) led to as much as a 12-fold increase in 3-ABA-derived DNA adduct formation compared with controls. The observed stimulation of DNA adduct formation by both compounds was attributed to their potential to induce protein expression and enzymatic activity of cytochromes P450 1A1 and/or -1A2 (CYP1A1/2), the major enzymes responsible for 3-ABA activation in human and rat livers. Collectively, these results demonstrate for the first time, to our knowledge, that by inducing hepatic NQO1 and CYP1A1/2, both 3-NBA and 3-ABA increase the enzymatic activation of these two compounds to reactive DNA adduct-forming species, thereby enhancing their own genotoxic potential. PMID- 16714373 TI - Quantitative determination of metabolic products of 19-norandrostenediol in human plasma using gas chromatography/mass spectrometry. AB - Prohormones such as 19-norandrostenediol (estr-4-ene-3beta,17beta-diol) have been added to the list of prohibited substances of the World Anti-Doping Agency because they are metabolized to the common nandrolone metabolites norandrosterone and noretiocholanolone. So far, no studies on the metabolism and in vivo conversion of 19-norandrostenediol after oral or sublingual administration have been reported nor have had quantified data on resulting plasma nandrolone levels. In the present study, an open-label crossover trial with eight healthy male volunteers was conducted. After application of capsules or sublingual tablets of 19-norandrostenediol plasma concentrations of 19-norandrostenediol, nandrolone as well as major metabolites (19-norandrosterone and 19-noretiocholanolone) were determined using a validated assay based on gas chromatography/mass spectrometry. The administration of 100-mg capsules of 19-norandrostenediol yielded maximum plasma total concentrations (i.e., conjugated plus unconjugated compounds) of 1.1 ng/ml (+/-0.7) for 19-norandrostenediol, 4.0 ng/ml (+/-2.6) for nandrolone, 154.8 ng/ml (+/-130.8) for 19-norandrosterone, and 37.7 ng/ml (+/-6.9) for 19 noretiocholanolone. The use of 25-mg sublingual tablets resulted in 3.3 ng/ml (+/ 1.0) for 19-norandrostenediol, 11.0 ng/ml (+/-6.4) for nandrolone, 106.3 ng/ml (+/-40.1) for 19-norandrosterone, and 28.5 ng/ml (+/-20.8) for 19 noretiocholanolone. Most interestingly, the pharmacologically active unconjugated nandrolone was determined after administration of sublingual tablets (up to 5.7 ng/ml) in contrast to capsule applications. These results demonstrate the importance of prohibiting prohormones such as 19-norandrostenediol, in particular, since plasma concentrations of nandrolone between 0.3 to 1.2 ng/ml have been reported to influence endocrinological parameters. PMID- 16714374 TI - Pharmacokinetics and metabolism of the flavonoid scutellarin in humans after a single oral administration. AB - Scutellarin is widely used in treating various cardiovascular diseases. Few data are available regarding its metabolism and pharmacokinetics in humans. The objectives of this study were to develop methods to identify major metabolites of scutellarin in human urine and plasma and to determine simultaneously the parent drug and its major metabolites in human plasma for pharmacokinetic studies. Four metabolites were detected in urine samples by liquid chromatography coupled with electrospray multi-stage mass spectrometry (MS), but only one of them was found in plasma. Its structure was confirmed as scutellarein 6-O-beta-D-glucuronide by MS, NMR, and UV absorbance spectra. The plasma concentrations of scutellarin and the major metabolite were simultaneously determined using liquid chromatography tandem MS. After a single p.o. administration of 60 mg of scutellarin to 20 healthy subjects, the plasma concentrations of scutellarin were very low, and its plasma concentration-time curve was also anomalous. Plasma concentration of the major metabolite was comparatively high, and the peak plasma concentration was 87.0 +/- 29.1 ng/ml. The Tmax was late (7.85 +/- 1.62 h), and part of individual pharmacokinetic profiles showed double peaks, which indicated scutellarin could be absorbed into the intestine after hydrolysis to its aglycone by bacterial enzymes. This was followed by reconjugation in the intestinal cell and/or liver with glucuronic acid catalyzed by the phase II enzyme, which showed regioselectivity and species difference. The regioselectivity of glucuronoconjugation for scutellarin may be of importance for pharmacological activity. Plasma concentration of isoscutellarin can be used as a biomarker of scutellarin intake. PMID- 16714375 TI - Intravenously administered short interfering RNA accumulates in the kidney and selectively suppresses gene function in renal proximal tubules. AB - Different gene-silencing methods, like antisense and short interfering RNA (siRNA), are widely used as experimental tools to inhibit gene expression. In the present study, the in vivo behavior of siRNA in rats and siRNA-mediated silencing of genes in the renal proximal tubule were investigated. To study the biodistribution of siRNA, rats were injected i.v. with radiolabeled siRNA or radiolabel alone (control), and scintigraphic images were acquired at different time intervals postinjection. The siRNA preferentially accumulated in the kidneys and was excreted in the urine. One hour after injection, the amount of siRNA present in both kidneys (1.7 +/- 0.3% of injected dose/g tissue) was on average 40 times higher than in other tissues (liver, brain, intestine, muscle, lung, spleen, and blood). Besides the biodistribution, the effect of siRNA on multidrug resistance protein isoform 2 (Mrp2/Abcc2, siRNAMrp2) in renal proximal tubules was investigated. Mrp2 function was assessed by measuring the excretion of its fluorescent substrate calcein in the isolated perfused rat kidney. Four days after administration, siRNAMrp2 reduced the urinary calcein excretion rate significantly (35% inhibition over the period 80-150 min of perfusion). This down regulation was specific because another siRNA sequence directed against a different transporter in the proximal tubule, Mrp4 (Abcc4, siRNAMrp4), did not alter the Mrp2-mediated excretion of calcein. In conclusion, siRNA accumulates spontaneously in the kidney after i.v. injection, where it selectively suppresses gene function in the proximal tubules. Therefore, i.v. administered siRNA provides a novel experimental and potential therapeutic tool for gene silencing in the kidney. PMID- 16714376 TI - Predominant contribution of organic anion transporting polypeptide OATP-B (OATP2B1) to apical uptake of estrone-3-sulfate by human intestinal Caco-2 cells. AB - Human organic anion transporting polypeptide OATP-B (OATP2B1) is a pH-sensitive transporter expressed in the apical membranes of small intestinal epithelial cells. In this study, we have examined the contribution of OATP-B to the uptake of [3H]estrone-3-sulfate in Caco-2 cells in comparison with those of its homologs OATP-D (OATP3A1) and OATP-E (OATP4A1). Immunocytochemical study revealed that OATP-B is expressed in the apical membranes of Caco-2 cells. The uptake of [3H]estrone-3-sulfate by Caco-2 cells was Na+-independent and inhibited by several organic anions. It showed biphasic saturation kinetics with Km values of 1.81 microM and 1.40 mM. The uptake of [3H]estrone-3-sulfate by human embryonic kidney (HEK) 293 cells stably expressing OATP-B (HEK293/OATP-B) was also Na+ independent and inhibited by several organic anions. The Km value for estrone-3 sulfate uptake by OATP-B (1.56 microM) was close to that for the high-affinity component observed in Caco-2 cells. The mRNA expression level of OATP-B was higher than that of OATP-D or OATP-E in Caco-2 cells and in human jejunum biopsies from healthy volunteers. The values of [3H]estrone-3-sulfate uptake normalized to OATP-B mRNA expression were similar in Caco-2 cells and HEK293/OATP B cells. The specific activity of OATP-B per mRNA expression was much higher than that of OATP-D and OATP-E. [3H]Estrone-3-sulfate uptake by membrane vesicles prepared from HEK293/OATP-B cells exhibited an overshoot phenomenon in the presence of an inwardly directed H+ gradient, suggesting that an H+ gradient is the driving force of estrone-3-sulfate transport by OATP-B. These results suggest that OATP-B is predominantly responsible for the apical uptake of estrone-3 sulfate in Caco-2 cells. PMID- 16714377 TI - Differential effects of the oxidized metabolites of oltipraz on the activation of CCAAT/enhancer binding protein-beta and NF-E2-related factor-2 for GSTA2 gene induction. AB - Comprehensive mechanistic studies suggest that oltipraz exerts cancer chemopreventive effects through the induction of glutathione S-transferase (GST). Previously, we have shown that the activation of CCAAT/enhancer binding protein beta (C/EBPbeta), promoted by oltipraz, contributes to the transcriptional induction of the GSTA2 gene. Studies also indicated that exposure of animals to oltipraz triggers nuclear accumulation of NF-E2-related factor-2 (Nrf2) with an increase in Nrf2's antioxidant response element (ARE) binding activity. Given the previous reports that C/EBPbeta activation contributes to oltipraz's induction of the GSTA2 gene and that Nrf2 activation by oltipraz was variable depending on the concentrations, this study investigated whether the major oxidized metabolites of oltipraz induce GSTA2 through the activation of C/EBPbeta and/or Nrf2. Immunoblot analysis revealed that M1 [4-methyl-5-(pyrazin-2-yl)-3H-1,2-dithiol-3-one] and M2 (7-methyl-6,8-bis(methylthio)H-pyrrolo[1,2-a]pyrazine), but not M3 (7-methyl-8 (methylsulfinyl)-6-(methylthio)H-pyrrolo[1,2-a]pyrazine) and M4 (7-methyl-6,8 bis(methylsulfinyl)H-pyrrolo[1,2-a]pyrazine), induced GSTA2 in H4IIE cells. M1 and M2 also increased the luciferase activity from pGL-1651, which contained the luciferase structural gene downstream of the -1.65-kilobase GSTA2 promoter region. Nuclear C/EBPbeta levels were enhanced by the metabolites but not by M3 or M4. Among the oxidized metabolites examined, only M2, which elicited cell death at a relatively high concentration, activated Nrf2, as indicated by nuclear accumulation of Nrf2 and its ARE binding activity. The present study provides evidence that M1 and M2, but not M3 and M4, induce GSTA2 and that M1 induces GSTA2 only via C/EBPbeta activation, whereas M2 does so by activating Nrf2 as well as C/EBPbeta. These results substantiate the differential effects of oltipraz's metabolites on C/EBPbeta- and/or Nrf2-mediated GSTA2 induction. PMID- 16714378 TI - Proposal to adapt the WASP programme to fibre counting tests. PMID- 16714380 TI - Distinct metastable atmospheric regimes despite nearly Gaussian statistics: a paradigm model. AB - A controversial topic in the recent climate modeling literature is the fashion in which metastable low-frequency regimes in the atmosphere occur despite nearly Gaussian statistics for these planetary waves. Here a simple 57-mode paradigm model for such metastable atmospheric regime behavior is introduced and analyzed through hidden Markov model (HMM) analysis of the time series of suitable low frequency planetary waves. The analysis of this paradigm model elucidates how statistically significant metastable regime transitions between blocked and zonal statistical states occur despite nearly Gaussian behavior in the associated probability distribution function and without a significant role for the low order truncated nonlinear dynamics alone; turbulent backscatter onto the three dimensional subspace of low-frequency modes is responsible for these effects. It also is demonstrated that suitable stochastic mode reduction strategies, which include both augmented cubic nonlinearity and multiplicative noise, are also capable of capturing the metastable low-frequency regime behavior through a single stochastic differential equation compared with the full turbulent chaotic 57-mode model. This feature is attractive for issues such as long-term weather predictability. Although there have been many applications of HMM in other sciences, this work presents a previously undescribed application of HMM analysis to atmospheric low-frequency variability and points the way for further applications including their use in extended range predictability. PMID- 16714379 TI - Essential role of mda-5 in type I IFN responses to polyriboinosinic:polyribocytidylic acid and encephalomyocarditis picornavirus. AB - The innate immune system recognizes viral dsRNA through two distinct pathways; the Toll-like receptor 3 (TLR3) pathway detects dsRNA phagocytosed in endosomes; the helicases retinoic acid-induced protein I (RIG-I) and melanoma differentiation-associated gene-5 (mda-5) detect cytoplasmic dsRNA generated during viral replication. Both RIG-I and mda-5 can bind polyriboinosinic:polyribocytidylic acid (polyI:C), the synthetic analog of viral dsRNA, and mediate type I IFN responses to polyI:C and multiple RNA viruses in vitro. We generated mda-5-deficient mice and showed that mda-5 is the dominant receptor mediating type I IFN secretion in response to polyI:C in vitro and in vivo. Moreover, mda-5-/- mice exhibited a selectively impaired antiviral response to encephalomyocarditis picornavirus, indicating functional specialization of mda 5 in vivo. PMID- 16714381 TI - p63 heterozygous mutant mice are not prone to spontaneous or chemically induced tumors. AB - Homology between p63 and p53 has suggested that these proteins might function similarly. However, the majority of data from human tumors have not supported a similar role for p63 in tumor suppression. To investigate this issue, we studied spontaneous tumorigenesis in p63+/- mice in both WT and p53-compromised backgrounds. We found that p63+/- mice were not tumor prone and mice heterozygous for both p63 and p53 had fewer tumors than p53+/- mice. The rare tumors that developed in mice with compromised p63 were also distinct from those of p53+/- mice. Furthermore, p63+/- mice were not prone to chemically induced tumorigenesis, and p63 expression was maintained in carcinomas. These findings demonstrate that, in agreement with data from human tumors, p63 plays a markedly different biological role in cancer than p53. PMID- 16714382 TI - Evidence of hollow golden cages. AB - The fullerenes are the first "free-standing" elemental hollow cages identified by spectroscopy experiments and synthesized in the bulk. Here, we report experimental and theoretical evidence of hollow cages consisting of pure metal atoms, Au(n)(-) (n = 16-18); to our knowledge, free-standing metal hollow cages have not been previously detected in the laboratory. These hollow golden cages ("bucky gold") have an average diameter >5.5 A, which can easily accommodate one guest atom inside. PMID- 16714383 TI - Hepatocyte nuclear factor 4alpha orchestrates expression of cell adhesion proteins during the epithelial transformation of the developing liver. AB - Epithelial formation is a central facet of organogenesis that relies on intercellular junction assembly to create functionally distinct apical and basal cell surfaces. How this process is regulated during embryonic development remains obscure. Previous studies using conditional knockout mice have shown that loss of hepatocyte nuclear factor 4alpha (HNF4alpha) blocks the epithelial transformation of the fetal liver, suggesting that HNF4alpha is a central regulator of epithelial morphogenesis. Although HNF4alpha-null hepatocytes do not express E cadherin (also called CDH1), we show here that E-cadherin is dispensable for liver development, implying that HNF4alpha regulates additional aspects of epithelial formation. Microarray and molecular analyses reveal that HNF4alpha regulates the developmental expression of a myriad of proteins required for cell junction assembly and adhesion. Our findings define a fundamental mechanism through which generation of tissue epithelia during development is coordinated with the onset of organ function. PMID- 16714384 TI - A bifunctional tRNA import receptor from Leishmania mitochondria. AB - In kinetoplastid protozoa, import of cytosolic tRNAs into mitochondria occurs through tRNAs interacting with membrane-bound proteins, the identities of which are unknown. The inner membrane RNA import complex of Leishmania tropica contains multiple proteins and is active for import in vitro. RIC1, the largest subunit of this complex, is structurally homologous to the conserved alpha subunit of F1 ATP synthase. The RIC1 gene complemented an atpA mutation in Escherichia coli. Antisense-mediated knockdown of RIC1/F1alpha in Leishmania resulted in depletion of several mitochondrial tRNAs belonging to distinct subsets (types I and II) that interact cooperatively or antagonistically within the import complex. The knockdown-induced defect in import of type I tRNAs was rectified in a reconstituted system by purified RIC1/F1alpha alone, but recovery of type II tRNA import additionally required a type I tRNA. RIC1/F1alpha formed stable complexes with type I, but not type II, tRNAs through the cooperation of its nucleotide binding and C-terminal domains. Thus, RIC1/F1alpha is a type I tRNA import receptor. As expected of a bifunctional protein, RIC1/F1alpha is shared by both the import complex and by respiratory complex V. Alternative use of ancient respiratory proteins may have been an important step in the evolution of tRNA import. PMID- 16714385 TI - Stochastic models for regulatory networks of the genetic toggle switch. AB - Bistability arises within a wide range of biological systems from the lambda phage switch in bacteria to cellular signal transduction pathways in mammalian cells. Changes in regulatory mechanisms may result in genetic switching in a bistable system. Recently, more and more experimental evidence in the form of bimodal population distributions indicates that noise plays a very important role in the switching of bistable systems. Although deterministic models have been used for studying the existence of bistability properties under various system conditions, these models cannot realize cell-to-cell fluctuations in genetic switching. However, there is a lag in the development of stochastic models for studying the impact of noise in bistable systems because of the lack of detailed knowledge of biochemical reactions, kinetic rates, and molecular numbers. In this work, we develop a previously undescribed general technique for developing quantitative stochastic models for large-scale genetic regulatory networks by introducing Poisson random variables into deterministic models described by ordinary differential equations. Two stochastic models have been proposed for the genetic toggle switch interfaced with either the SOS signaling pathway or a quorum-sensing signaling pathway, and we have successfully realized experimental results showing bimodal population distributions. Because the introduced stochastic models are based on widely used ordinary differential equation models, the success of this work suggests that this approach is a very promising one for studying noise in large-scale genetic regulatory networks. PMID- 16714387 TI - Strategies for the treatment of sepsis. PMID- 16714386 TI - Differential regulation of metabolic, neuroendocrine, and immune function by leptin in humans. AB - To elucidate whether the role of leptin in regulating neuroendocrine and immune function during short-term starvation in healthy humans is permissive, i.e., occurs only when circulating leptin levels are below a critical threshold level, we studied seven normal-weight women during a normoleptinemic-fed state and two states of relative hypoleptinemia induced by 72-h fasting during which we administered either placebo or recombinant methionyl human leptin (r-metHuLeptin) in replacement doses. Fasting for 72 h decreased leptin levels by approximately = 80% from a midphysiologic (14.7 +/- 2.6 ng/ml) to a low-physiologic (2.8 +/- 0.3 ng/ml) level. Administration of r-metHuLeptin during fasting fully restored leptin to physiologic levels (28.8 +/- 2.0 ng/ml) and reversed the fasting associated decrease in overnight luteinizing hormone pulse frequency but had no effect on fasting-induced changes in thyroid-stimulating hormone pulsatility, thyroid and IGF-1 hormone levels, hypothalamic-pituitary-adrenal and renin aldosterone activity. FSH and sex steroid levels were not altered. Short-term reduction of leptin levels decreased the number of circulating cells of the adaptive immune response, but r-metHuLeptin did not have major effects on their number or in vitro function. Thus, changes of leptin levels within the physiologic range have no major physiologic effects in leptin-replete humans. Studies involving more severe and/or chronic leptin deficiency are needed to precisely define the lower limit of normal leptin levels for each of leptin's physiologic targets. PMID- 16714389 TI - Variable in vivo embryoprotective role for ataxia-telangiectasia-mutated against constitutive and phenytoin-enhanced oxidative stress in atm knockout mice. AB - Knockout mice lacking the ataxia-telangiectasia-mutated (Atm) protein exhibit impaired detection and repair of DNA damage and increased embryopathies from ionizing radiation in vivo, and vehicle or phenytoin in embryo culture. Here we determined if Atm-deficient mice are more susceptible in vivo to phenytoin embryopathies. Wild-type (+/+) or heterozygous (+/-) Atm knockout dams were mated with +/- males, pregnant dams were treated with phenytoin (65 mg/kg ip) or its vehicle, and resorptions and fetuses were genotyped and characterized. This strain proved resistant to phenytoin-initiated cleft palates but not to other spontaneous and phenytoin-enhanced embryopathies. With vehicle-treated +/- dams, fetal body weight was lower in homozygous Atm-null (-/-) fetuses compared to +/- and +/+ littermates (p < 0.05). Phenytoin enhanced this Atm-dependent embryopathic pattern (p < 0.05). It also enhanced DNA oxidation in -/- Atm deficient embryos compared to its +/- Atm-deficient (p < 0.001) and +/+ Atm normal (p < 0.001), phenytoin-exposed littermates and to its -/- vehicle controls (p < 0.01). Postpartum lethality was greater in both +/- and -/- Atm-deficient fetuses compared to +/+ littermates, independent of treatment (0.05 < p < 0.1). By maternal genotype, +/- Atm-deficient dams had fewer implantations than +/+ dams, independent of treatment, and phenytoin decreased litter size (p < 0.05). Conversely, phenytoin-exposed +/+ fetuses were more likely than -/- littermates to die in utero (p < 0.05), and in +/+ dams fetal resorptions and postpartum lethality were variably higher and enhanced by phenytoin (p < 0.05). Despite variable actions in vivo, the embryoprotective effects of Atm suggest a role for reactive oxygen species and oxidative DNA damage in some spontaneous and phenytoin-enhanced embryopathies. PMID- 16714388 TI - Targeting of the plant vacuolar sorting receptor BP80 is dependent on multiple sorting signals in the cytosolic tail. AB - Although signals for vacuolar sorting of soluble proteins are well described, we have yet to learn how the plant vacuolar sorting receptor BP80 reaches its correct destination and recycles. To shed light on receptor targeting, we used an in vivo competition assay in which a truncated receptor (green fluorescent protein-BP80) specifically competes with sorting machinery and causes hypersecretion of BP80-ligands from tobacco (Nicotiana tabacum) leaf protoplasts. We show that both the transmembrane domain and the cytosolic tail of BP80 contain information necessary for efficient progress to the prevacuolar compartment (PVC). Furthermore, the tail must be exposed on the correct membrane surface to compete with sorting machinery. Mutational analysis of conserved residues revealed that multiple sequence motifs are necessary for competition, one of which is a typical Tyr-based motif (YXXPhi). Substitution of Tyr-612 for Ala causes partial retention in the Golgi apparatus, mistargeting to the plasma membrane (PM), and slower progress to the PVC. A role in Golgi-to-PVC transport was confirmed by generating the corresponding mutation on full-length BP80. The mutant receptor was partially mistargeted to the PM and induced the secretion of a coexpressed BP80-ligand. Further mutants indicate that the cytosolic tail is likely to contain other information besides the YXXPhi motif, possibly for endoplasmic reticulum export, endocytosis from the PM, and PVC-to-Golgi recycling. PMID- 16714390 TI - XRCC1 protects against particulate chromate-induced chromosome damage and cytotoxicity in Chinese hamster ovary cells. AB - Water-insoluble hexavalent chromium compounds are well-established human lung carcinogens. Lead chromate, a model insoluble Cr(VI) compound, induces DNA damage, chromosome aberrations, and dose-dependent cell death in human and Chinese hamster ovary (CHO) cells. The relationship between lead chromate-induced DNA damage and chromosome aberrations is unknown. Our study focus was on examining the role of XRCC1 in lead chromate-induced cytotoxicity and structural chromosomal aberrations in CHO cells. Three different cell lines were used: AA8 (parental), EM9 (XRCC1 mutant), and H9T3 (EM9 complemented with human XRCC1 gene). Cytotoxicity was significantly higher in EM9 cells when compared to AA8 and H9T3 cells, indicating that XRCC1 is important for protecting cells from lead chromate particles-induced cell death. The frequency of damaged metaphase cells was not affected by XRCC1 deficiency. However, the total amount of Cr(VI)-induced chromosome damage was exacerbated by XRCC1 deficiency, and the spectrum of damage changed dramatically. Chromatid and isochromatid lesions were the most prominent aberrations induced in all cell lines. XRCC1 was essential to reduce the formation of chromatid lesions but not for isochromatid lesions. In addition, XRCC1 deficiency resulted in a dramatic increase in the number of chromatid exchanges, indicating that XRCC1 is involved in protection from lead chromate induced chromosome instability. PMID- 16714391 TI - The interaction of manganese nanoparticles with PC-12 cells induces dopamine depletion. AB - This investigation was designed to determine whether nano-sized manganese oxide (Mn-40 nm) particles would induce dopamine (DA) depletion in a cultured neuronal phenotype, PC-12 cells, similar to free ionic manganese (Mn(2+)). Cells were exposed to Mn-40 nm, Mn(2+) (acetate), or known cytotoxic silver nanoparticles (Ag-15 nm) for 24 h. Phase-contrast microscopy studies show that Mn-40 nm or Mn(2+) exposure did not greatly change morphology of PC-12 cells. However, Ag-15 nm and AgNO(3) produce cell shrinkage and irregular membrane borders compared to control cells. Further microscopic studies at higher resolution demonstrated that Mn-40 nm nanoparticles and agglomerates were effectively internalized by PC-12 cells. Mitochondrial reduction activity, a sensitive measure of particle and metal cytotoxicity, showed only moderate toxicity for Mn-40 nm compared to similar Ag-15 nm and Mn(2+) doses. Mn-40 nm and Mn(2+) dose dependently depleted DA and its metabolites, dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), while Ag-15 nm only significantly reduced DA and DOPAC at concentrations of 50 mug/ml. Therefore, the DA depletion of Mn-40 nm was most similar to Mn(2+), which is known to induce concentration-dependent DA depletion. There was a significant increase (> 10-fold) in reactive oxygen species (ROS) with Mn-40 nm exposure, suggesting that increased ROS levels may participate in DA depletion. These results clearly demonstrate that nanoscale manganese can deplete DA, DOPAC, and HVA in a dose-dependent manner. Further study is required to evaluate the specific intracellular distribution of Mn-40 nm nanoparticles, metal dissolution rates in cells and cellular matrices, if DA depletion is induced in vivo, and the propensity of Mn nanoparticles to cross the blood-brain barrier or be selectively uptaken by nasal epithelium. PMID- 16714393 TI - New ventilators for the ICU--usefulness of lung performance reporting. AB - Monitoring the functional and mechanical properties of the lungs during positive pressure ventilation may assist in confirming the underlying pulmonary diagnosis, allow therapeutic interventions to be accurately assessed and provide information that ensures the optimal setting of the ventilator parameters and encourages timely weaning. This article reviews the range of lung function measurements, both continuous and intermittent, that may be undertaken during mechanical ventilation. The monitoring capability of ICU ventilators is increasing in complexity. PMID- 16714392 TI - Functional analysis of a metal response element in the regulatory region of flounder cytochrome P450 1A and implications for environmental monitoring of pollutants. AB - Cytochrome P450 1A (CYP 1A) is a member of a multigene family of xenobiotic metabolizing enzymes. CYP 1A is highly inducible by numerous environmental contaminants including polycyclic aromatic hydrocarbons (PAHs) and is widely used in biomonitoring studies. Therefore, understanding the regulation of this gene is important for accurate interpretation of biomarker data. We describe here the functional role of a metal response element (MRE) in the European flounder CYP 1A promoter region. To help elucidate the potential role of this MRE, reporter gene constructs, with or without site-directed mutagenesis, were used in conjunction with a dual-luciferase assay. The electrophoretic mobility shift assay (EMSA) was also used to investigate potential protein binding at this MRE site. Treatment with the prototypical PAH 3-methylcholanthrene (3MC) (1.0 microM) produced a dose dependent response at the CYP 1A promoter, whereas treatment with cadmium (0-1.0 microM) produced little transcriptional activity at either the wild-type or mutated promoter. Cotreatment with cadmium (1.0 microM) and 3MC (1.0 microM) reduced induction at this promoter to 1.83-fold compared to 3MC treatment alone (4.0-fold induction). Mutation of the MRE site resulted in abolishment of this cadmium-related loss of 3MC-dependent activity. Furthermore, a retarded band was observed in the EMSA when the MRE was used as a probe and incubated with liver nuclear protein from flounder treated with cadmium. The results not only add to knowledge of the diversity in vertebrate CYP 1A regulation but also raise the complexity of interpretation of CYP 1A induction in monitoring studies that involve mixtures of PAHs and metals. PMID- 16714395 TI - Pathobiological characteristics of intestinal and diffuse-type gastric carcinoma in Japan: an immunostaining study on the tissue microarray. AB - AIM: To investigate the pathobiological features of intestinal and diffuse-type gastric carcinomas in the Japanese population. METHODS: The expression of fragile histine triad (FHIT), phosphatase and tensin homology deleted from human chromosome 10 (PTEN), caspase-3, Ki-67, mutant p53, matrix metalloproteinase (MMP)-2, MMP-9, and extracellular matrix metalloproteinase inducer (EMMPRIN) on tissue microarrays of gastric carcinomas by immunostaining was examined in comparison with the clinicopathological characteristics between intestinal and diffuse-type cases. RESULTS: Intestinal-type carcinoma frequently occurred in old men, whereas the diffuse type comparatively occurred more in young women (p<0.05). The diffuse-type carcinoma was more inclined to invasion into muscularis propria, lymphatic invasion and lymph node metastasis, and belonged to higher International Union against Cancer (UICC) staging (p<0.05) compared with intestinal-type counterparts. Expression of FHIT, PTEN, Ki-67, caspase-3, mutant p53 and EMPPRIN was higher in intestinal-type carcinomas than in diffuse-type carcinomas (p<0.05). Kaplan-Meier analysis indicated that patients with intestinal-type carcinomas had a higher cumulative survival rate (p<0.05). CONCLUSION: Intestinal-type gastric carcinomas with a more favourable prognosis frequently show high levels of proliferation and apoptosis, and always accompany strong expression of FHIT, PTEN and mutant p53 and EMMPRIN. EMMPRIN expression might underlie the molecular basis of liver metastasis and higher proliferation of intestinal-type gastric carcinomas in Japan. Lauren's classification thus proved pathologically relevant for the clinical treatment of gastric carcinomas. PMID- 16714396 TI - Survivin promoter polymorphism and cervical carcinogenesis. AB - BACKGROUND: Survivin, a novel member of the inhibitor of apoptosis family, plays an important role in cell cycle regulation. A common polymorphism at the survivin gene promoter (G/C at position 31) was shown to be correlated with survivin gene expression in cancer cell lines. AIM: To investigate whether this polymorphism could be involved in the development of human papillomavirus (HPV)-associated cervical carcinoma. METHODS: Survivin promoter polymorphism was detected in patients with cervical cancer, in patients with equivocal cytological atypia and in a control population using polymerase chain reaction (PCR-restriction fragment length polymorphism (RFLP) and PCR-single strand conformation polymorphism analysis. HPV was typed in patients with cervical cancer and cytological atypia using PCR-RFLP. RESULTS: No statistically significant differences were found in the genotype distributions of the survivin promoter variants among our study groups. CONCLUSIONS: The survivin promoter polymorphism at position 31 may not represent an increased risk for the development of cervical cancer, at least in the population studied here. PMID- 16714398 TI - A survey of time management and particular tasks undertaken by consultant microbiologists in the UK. AB - BACKGROUND: Medical microbiology practice encompasses a diverse range of activities. Consultant medical microbiologists (CMMs) attribute widely differing priorities to, and spend differing proportions of time on various components of the job. AIM: To obtain a professional consensus on what are high-priority and low-priority activities, and to identify the time spent on low-priority activities. METHOD: National survey. RESULTS: Many respondents felt that time spent on report authorisation and telephoning of results was excessive, whereas time spent on ward-based work was inadequate. Timesaving could also be achieved through better prioritisation of infection-control activities. CONCLUSION: CMMs should apportion their time at work focusing on high-priority activities identified through professional consensus. PMID- 16714400 TI - Policosanol inhibits cholesterol synthesis in hepatoma cells by activation of AMP kinase. AB - Policosanol is a mixture of long-chain primary alcohols that has been shown to decrease serum cholesterol in animals and in humans. The hypocholesterolemic effect results from a decrease in cholesterol synthesis by suppression of HMG-CoA reductase activity, but the mechanism of this suppression and the active components of policosanol have not been established. In the present study, we investigated the ability of policosanol and its principal components to inhibit cholesterol synthesis in cultured rat hepatoma cells. Maximal inhibition by policosanol yielded a 30% decrease in [(14)C]acetate incorporation without evidence of cellular toxicity. Octacosanol (C28, the major constituent of policosanol), heptacosanol (C27), and hexacosanol (C26) yielded smaller and statistically insignificant decreases in cholesterol synthesis, whereas triacontanol (1-hydroxytriacontane; C30) replicated the inhibition obtained with policosanol. At pharmacological concentrations (<5 microg/ml), policosanol and triacontanol decreased [(14)C]acetate incorporation into cholesterol without affecting the incorporation of [(14)C]mevalonate, indicating that these compounds act at or above HMG-CoA reductase. Policosanol and triacontanol did not directly inhibit HMG-CoA reductase, and incubation of these compounds with hepatoma cells did not affect reductase enzyme levels. However, reductase activity was decreased by up to 55% in lysates prepared from these cells, suggesting that HMG-CoA reductase activity was down-regulated by policosanol treatment. Consistent with this hypothesis, a 3-fold increase in AMP-kinase phosphorylation was noted in policosanol-treated cells. Because AMP-kinase is activated by phosphorylation and is well established to suppress HMG-CoA reductase activity, these results suggest that policosanol or a metabolite decreases HMG-CoA reductase activity by activating AMP-kinase. PMID- 16714397 TI - Best practice in primary care pathology: review 4. AB - This fourth best practice review examines four series of common primary care questions in laboratory medicine are examined in this review: (1) safety monitoring for three common drugs; (2) use of prostate-specific antigen; (3) investigation of vaginal discharge; and (4) investigation of subfertility. The review is presented in question-answer format, referenced for each question series. The recommendations represent a precis of the guidance found using a standardised literature search of national and international guidance notes, consensus statements, health policy documents and evidence-based medicine reviews, supplemented by Medline Embase searches to identify relevant primary research documents. They are not standards but form a guide to be set in the clinical context. Most of them are consensus based rather than evidence based. They will be updated periodically to take account of new information. PMID- 16714401 TI - Tachykinin receptor expression and function in human esophageal smooth muscle. AB - Tachykinins are present in enteric nerves of the gastrointestinal tract and cause contraction of esophageal smooth muscle; however, the mechanisms involved are not understood. Our aim was to characterize tachykinin signaling in human esophageal smooth muscle. We investigated functional effects of tachykinins on human esophageal smooth muscle using tension recordings and isolated cells, receptor expression with reverse transcription (RT)-polymerase chain reaction (PCR) and immunoblotting, intracellular Ca2+ responses using fluorescent indicator dyes, and membrane currents with patch-clamp electrophysiology. The mammalian tachykinins [substance P and neurokinin (NK) A and NKB] elicited concentration dependent contractions of human esophageal smooth muscle. These responses were not affected by muscarinic receptor or neuronal blockade indicating a direct effect on smooth muscle cells (SMCs). Immunofluorescence and RT-PCR identified tachykinin receptors (NK1, NK2, and NK3) on SMCs. Contraction was mediated through a combination of Ca2+ release from intracellular stores and influx through L-type Ca2+ channels. NK2 receptor blockade inhibited the largest proportion of tachykinin-evoked responses. NKA evoked a nonselective cation current (I(NSC)) with properties similar to that elicited by muscarinic stimulation. The following paradigm is suggested: tachykinin receptor binding to SMCs releases Ca2+ from stores along with activation of I(NSC), which in turn results in membrane depolarization, L-type Ca2+ channel opening, rise of Ca2+ concentration, and contraction. These studies reveal new aspects of tachykinin signaling in human esophageal SMCs. Excitatory tachykinin pathways may represent targets for pharmacological intervention in disorders of esophageal dysmotility. PMID- 16714399 TI - In the absence of (early) invasive carcinoma, vulvar intraepithelial neoplasia associated with lichen sclerosus is mainly of undifferentiated type: new insights in histology and aetiology. AB - BACKGROUND: Differentiated vulvar intraepithelial neoplasia (VIN) is presumed to be the precursor of invasive squamous cell carcinoma (SCC) of the vulva. It is commonly assumed that differentiated VIN is related to lichen sclerosus (LS). However, evidence for this is limited to a small number of studies describing epithelial alterations adjacent to vulvar SCC. AIM: To study the histology and human papillomavirus (HPV) status in patients with a history of both LS and VIN without coexistent SCC. METHODS: Original biopsy specimens and surgical specimens of patients retrieved from the pathology files were revised for the presence of LS, VIN and (early) invasive SCC, specifically focused on the two different types of VIN: differentiated and undifferentiated. Thereafter, VIN lesions were tested for the presence of HPV DNA. RESULTS: Twenty-seven patients fulfilled the criteria for LS and VIN without SCC. In all 27 patients, LS was found to be related to undifferentiated VIN. Grading yielded the following results: VIN 1 (n=10), VIN 2 (n=11) and VIN 3 (n=6). Additionally, VIN lesions from 26 patients could be tested for the presence of HPV DNA. HPV DNA, predominantly type 16, was present in 8 (31%) of them. Seven of these eight patients had VIN 2 or 3. During follow-up, three patients progressed to (early) invasive carcinoma. In two of these patients, differentiated VIN was observed overlying early invasive SCC. CONCLUSIONS: VIN related to LS without coexisting SCC is likely to be undifferentiated, in contrast to what was previously thought. HPV DNA was demonstrated in 31% of the lesions, and was strongly related to high-grade VIN. PMID- 16714402 TI - Effect of a novel vacuolar-H+-ATPase inhibitor on cell and tumor response to camptothecins. AB - The vacuolar-H(+)-ATPase, functionally expressed in cell membranes, is known to play a relevant role in intracellular pH regulatory mechanisms, because it is implicated in pumping protons into the extracellular environment or in sequestrating excess protons into acidic vacuolar compartments. Because tumor cells exist in a hypoxic microenvironment and produce acidic metabolites, this regulatory mechanism is recognized as a protective function. This study was designed to investigate the effect of NiK-12192 [4-(5,6-dichloro-1H-indol-2-yl)-3 ethoxy-N-(2,2,6,6-tetramethyl-piperidin-4-yl)-benzamide], an indole derivative identified as an effective inhibitor of vacuolar-H(+)-ATPase, on the cytotoxic activity of two camptothecins, i.e., topotecan and SN-38 (7-ethyl-10 hydroxycamptothecin, the active metabolite of irinotecan). The cellular studies performed in two pairs of human colon carcinoma cell lines, i.e., LoVo and LoVo/DX (overexpressing P-glycoprotein) and HT29 and HT29/Mit (overexpressing breast cancer resistant protein), indicated an enhancement of the antiproliferative effect of camptothecins by concomitant exposure to subtoxic concentrations of NiK-12192. Studies of subcellular distribution indicated that whereas topotecan alone localized mainly in mitochondria and endoplasmic compartment, the simultaneous presence of NiK-12192 caused a cytoplasmic redistribution. In HT29/Mit cells, NiK-12192 reverted the pattern of acidification induced by topotecan. The potentiation of topotecan efficacy by NiK 12192 was documented by an increased efficacy of the combination in both the HT29 tumor xenografts, being more evident in the topotecan-resistant HT29/Mit tumor. In conclusion, the vacuolar-H(+)-ATPase inhibitor NiK-12192 was able to potentiate the cytotoxic/antitumor effects of camptothecins, either in in vitro or in in vivo systems. Such findings support a potential interest for the use of vacuolar-H(+)-ATPase inhibitors in combination therapy to improve camptothecin efficacy. PMID- 16714403 TI - Drug-induced expression of nonsteroidal anti-inflammatory drug-activated gene/macrophage inhibitory cytokine-1/prostate-derived factor, a putative tumor suppressor, inhibits tumor growth. AB - A common in vitro response for many chemopreventive and antitumor agents, including some cyclooxygenase inhibitors, is the increased expression of nonsteroidal anti-inflammatory drug-activated gene (NAG)-1/macrophage inhibitory cytokine (MIC)-1/prostate-derived factor (PDF). The experimental anticancer drug 2-(4-amino-3-methylphenyl)-5-fluorobenzothiazole (5F203) was a potent inducer of NAG-1 expression, and in MCF-7 cells, it inhibited cell growth and induced apoptosis. NAG-1 small interfering RNA blocked NAG-1 expression and 5F203-induced apoptosis in MCF-7 cells, indicating that NAG-1 may mediate the apoptosis and anticancer activity. One mechanism by which 5F203 increases NAG-1 expression is by increasing the stability of NAG-1 mRNA, dependent of de novo protein synthesis. Extracellular signal-regulated kinase (ERK) 1/2 phosphorylation was increased by 5F203, and inhibition of ERK1/2 phosphorylation abolished the induction of NAG-1 protein expression and increased the stability of NAG-1 mRNA. Thus, 5F203 regulates NAG-1 expression by a unique mechanism compared with other drugs. A mouse orthotopic mammary tumor model was used to determine whether 5F203 increased NAG-1 expression in vivo and suppressed tumor growth. Treatment of the mice with Phortress, the prodrug of 5F203, increased the in vivo expression of NAG-1 as measured by real-time reverse transcription-polymerase chain reaction from RNA obtained by needle biopsy, and the expression correlated with a reduction of tumor volume. These results confirm that NAG-1 suppresses tumor growth, and its in vivo expression can be controlled by treating mice with anticancer drugs, such as Phortress. Drugs that target NAG-1 could lead to a unique strategy for the development of chemotherapeutic and chemopreventive agents. PMID- 16714404 TI - Comparative genomic analysis revealed a gene for monoglucosyldiacylglycerol synthase, an enzyme for photosynthetic membrane lipid synthesis in cyanobacteria. AB - Cyanobacteria have a thylakoid lipid composition very similar to that of plant chloroplasts, yet cyanobacteria are proposed to synthesize monogalactosyldiacylglycerol (MGDG), a major membrane polar lipid in photosynthetic membranes, by a different pathway. In addition, plant MGDG synthase has been cloned, but no ortholog has been reported in cyanobacterial genomes. We report here identification of the gene for monoglucosyldiacylglycerol (MGlcDG) synthase, which catalyzes the first step of galactolipid synthesis in cyanobacteria. Using comparative genomic analysis, candidates for the gene were selected based on the criteria that the enzyme activity is conserved between two species of cyanobacteria (unicellular [Synechocystis sp. PCC 6803] and filamentous [Anabaena sp. PCC 7120]), and we assumed three characteristics of the enzyme; namely, it harbors a glycosyltransferase motif, falls into a category of genes with unknown function, and shares significant similarity in amino acid sequence between these two cyanobacteria. By a motif search of all genes of Synechocystis, BLAST searches, and similarity searches between these two cyanobacteria, we identified four candidates for the enzyme that have all the characteristics we predicted. When expressed in Escherichia coli, one of the Synechocystis candidate proteins showed MGlcDG synthase activity in a UDP-glucose dependent manner. The ortholog in Anabaena also showed the same activity. The enzyme was predicted to require a divalent cation for its activity, and this was confirmed by biochemical analysis. The MGlcDG synthase and the plant MGDG synthase shared low similarity, supporting the presumption that cyanobacteria and plants utilize different pathways to synthesize MGDG. PMID- 16714405 TI - Mutagenic definition of a papain-like catalytic triad, sufficiency of the N terminal domain for single-site core catalytic enzyme acylation, and C-terminal domain for augmentative metal activation of a eukaryotic phytochelatin synthase. AB - Phytochelatin (PC) synthases are gamma-glutamylcysteine (gamma-Glu-Cys) dipeptidyl transpeptidases that catalyze the synthesis of heavy metal-binding PCs, (gamma-Glu-Cys)nGly polymers, from glutathione (GSH) and/or shorter chain PCs. Here it is shown through investigations of the enzyme from Arabidopsis (Arabidopsis thaliana; AtPCS1) that, although the N-terminal half of the protein, alone, is sufficient for core catalysis through the formation of a single-site enzyme acyl intermediate, it is not sufficient for acylation at a second site and augmentative stimulation by free Cd2+. A purified N-terminally hexahistidinyl tagged AtPCS1 truncate containing only the first 221 N-terminal amino acid residues of the enzyme (HIS-AtPCS1_221tr) is competent in the synthesis of PCs from GSH in media containing Cd2+ or the synthesis of S-methyl-PCs from S methylglutathione in media devoid of heavy metal ions. However, whereas its full length hexahistidinyl-tagged equivalent, HIS-AtPCS1, undergoes gamma-Glu-Cys acylation at two sites during the Cd2+-dependent synthesis of PCs from GSH and is stimulated by free Cd2+ when synthesizing S-methyl-PCs from S-methylglutathione, HIS-AtPCS1_221tr undergoes gamma-Glu-Cys acylation at only one site when GSH is the substrate and is not directly stimulated, but instead inhibited, by free Cd2+ when S-methylglutathione is the substrate. Through the application of sequence search algorithms capable of detecting distant homologies, work we reported briefly before but not in its entirety, it has been determined that the N terminal half of AtPCS1 and its equivalents from other sources have the hallmarks of a papain-like, Clan CA Cys protease. Whereas the fold assignment deduced from these analyses, which substantiates and is substantiated by the recent determination of the crystal structure of a distant prokaryotic PC synthase homolog from the cyanobacterium Nostoc, is capable of explaining the strict requirement for a conserved Cys residue, Cys-56 in the case of AtPCS1, for formation of the biosynthetically competent gamma-Glu-Cys enzyme acyl intermediate, the primary data from experiments directed at determining whether the other two residues, His-162 and Asp-180 of the putative papain-like catalytic triad of AtPCS1, are essential for catalysis have yet to be presented. This shortfall in our basic understanding of AtPCS1 is addressed here by the results of systematic site-directed mutagenesis studies that demonstrate that not only Cys-56 but also His-162 and Asp-180 are indeed required for net PC synthesis. It is therefore established experimentally that AtPCS1 and, by implication, other eukaryotic PC synthases are papain Cys protease superfamily members but ones, unlike their prokaryotic counterparts, which, in addition to having a papain-like N-terminal catalytic domain that undergoes primary gamma-Glu-Cys acylation, contain an auxiliary metal-sensing C-terminal domain that undergoes secondary gamma-Glu-Cys acylation. PMID- 16714406 TI - Quantitative trait loci analysis of primary cell wall composition in Arabidopsis. AB - Quantitative trait loci (QTL) analysis was used to identify genes underlying natural variation in primary cell wall composition in Arabidopsis (Arabidopsis thaliana). The cell walls of dark-grown seedlings of a Bay-0 x Shahdara recombinant inbred line population were analyzed using three miniaturized global cell wall fingerprinting techniques: monosaccharide composition analysis by gas chromatography, xyloglucan oligosaccharide mass profiling, and whole-wall Fourier transform infrared microspectroscopy. Heritable variation and transgression were observed for the arabinose-rhamnose ratio, xyloglucan side-chain composition (including O-acetylation levels), and absorbance for a subset of Fourier transform infrared wavenumbers. In total, 33 QTL, corresponding to at least 11 different loci controlling dark-grown hypocotyl length, pectin composition, and levels of xyloglucan fucosylation and O-acetylation, were identified. One major QTL, accounting for 51% of the variation in the arabinose-rhamnose ratio, affected the number of arabinan side chains presumably attached to the pectic polysaccharide rhamnogalacturonan I, paving the way to positional cloning of the first gene underlying natural variation in pectin structure. Several QTL were found to be colocalized, which may have implications for the regulation of xyloglucan metabolism. These results demonstrate the feasibility of combining fingerprinting techniques, natural variation, and quantitative genetics to gain original insight into the molecular mechanisms underlying the structure and metabolism of cell wall polysaccharides. PMID- 16714407 TI - Morphological alteration caused by brassinosteroid insensitivity increases the biomass and grain production of rice. AB - The rice (Oryza sativa) dwarf mutant d61 phenotype is caused by loss of function of a rice BRASSINOSTEROID INSENSITIVE1 ortholog, OsBRI1. We have identified nine d61 alleles, the weakest of which, d61-7, confers agronomically important traits such as semidwarf stature and erect leaves. Because erect-leaf habit is considered to increase light capture for photosynthesis, we compared the biomass and grain production of wild-type and d61-7 rice. The biomass of wild type was 38% higher than that of d61-7 at harvest under conventional planting density because of the dwarfism of d61-7. However, the biomass of d61-7 was 35% higher than that of wild type at high planting density. The grain yield of wild type reached a maximum at middensity, but the yield of d61-7 continued to increase with planting density. These results indicate that d61-7 produces biomass more effectively than wild type, and consequently more effectively assimilates the biomass in reproductive organ development at high planting density. However, the small grain size of d61-7 counters any increase in grain yield, leading to the same grain yield as that of wild type even at high density. We therefore produced transgenic rice with partial suppression of endogenous OsBRI1 expression to obtain the erect-leaf phenotype without grain changes. The estimated grain yield of these transformants was about 30% higher than that of wild type at high density. These results demonstrate the feasibility of generating erect-leaf plants by modifying the expression of the brassinosteroid receptor gene in transgenic rice plants. PMID- 16714408 TI - AINTEGUMENTA contributes to organ polarity and regulates growth of lateral organs in combination with YABBY genes. AB - Lateral organs in flowering plants display polarity along their adaxial-abaxial axis with distinct cell types forming at different positions along this axis. Members of three classes of transcription factors in Arabidopsis (Arabidopsis thaliana; the Class III homeodomain/leucine zipper [HD-ZIP] proteins, KANADI proteins, and YABBY proteins) are expressed in either the adaxial or abaxial domain of organ primordia where they confer these respective identities. Little is known about the factors that act upstream of these polarity-determining genes to regulate their expression. We have investigated the relationship between AINTEGUMENTA (ANT), a gene that promotes initiation and growth of lateral organ primordia, and polarity genes. Although ant single mutants do not display any obvious defects in organ polarity, loss of ANT activity in combination with mutations in one or more YABBY genes results in polarity defects greater than those observed in the yabby mutants alone. Our results suggest that ANT acts in combination with the YABBY gene FILAMENTOUS FLOWER (FIL) to promote organ polarity by up-regulating the expression of the adaxial-specifying HD-ZIP gene PHABULOSA. Furthermore, we show that ANT acts with FIL to up-regulate expression of the floral homeotic gene APETALA3. Our work defines new roles for ANT in the development of lateral organs. PMID- 16714409 TI - Aryl hydrocarbon receptor activation produces heart-specific transcriptional and toxic responses in developing zebrafish. AB - Proper regulation of the aryl hydrocarbon receptor (AHR), a ligand-activated transcription factor, is required for normal vertebrate cardiovascular development. AHR hyperactivation by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) during zebrafish (Danio rerio) development results in altered heart morphology and function, culminating in death. To identify genes that may cause cardiac toxicity, we analyzed the transcriptional response to TCDD in zebrafish hearts. Zebrafish larvae were exposed to TCDD for 1 h at 72 h after fertilization (hpf), and the hearts were extracted for microarray analysis at 1, 2, 4, and 12 h after exposure (73, 74, 76, and 84 h postfertilization). The remaining body tissue was also collected at each time for comparison. TCDD rapidly induced expression in 42 genes within 1 to2hof exposure. These genes function in xenobiotic metabolism, proliferation, heart contractility, and pathways that regulate heart development. Furthermore, these expression changes preceded signs of cardiovascular toxicity, characterized by decreased stroke volume, peripheral blood flow, and a halt in heart growth. This identifies strong candidates for important AHR target genes. It is noteworthy that the TCDD-induced transcriptional response in the hearts of zebrafish larvae was substantially different from that induced in the rest of the body tissues. One of the biggest differences included a cluster of genes that were down-regulated 12 h after exposure in heart tissue, but not in the body samples. More than 70% of the transcripts in this heart-specific cluster promote cellular growth and proliferation. Thus, the developing heart stands out as being responsive to TCDD at both the level of toxicity and gene expression. PMID- 16714410 TI - Supraspinal fatigue: the effects of caffeine on human muscle performance. PMID- 16714411 TI - It's all in the genes, so pick your parents wisely. PMID- 16714412 TI - Waste not, weak not? PMID- 16714413 TI - The many flavors of IGF-I. PMID- 16714414 TI - Chronic hypoxia augments uterine artery distensibility and alters the circumferential wall stress-strain relationship during pregnancy. AB - Pregnancy-associated increases in uterine artery (UA) blood flow are due, in part, to vasoactive and growth-related changes that enlarge UA diameter. Although active and passive mechanical factors can contribute to this enlargement, their role is less well understood. We hypothesized that pregnancy increased UA distensibility and/or decreased myogenic tone. Given the fetal growth restriction and lower UA flow seen under chronic hypoxia, we further hypothesized that chronic hypoxia opposed these normal active and passive mechanical changes. UA were isolated from 12 nonpregnant and 12 pregnant (0.7 gestation) guinea pigs housed under normoxia or chronic hypoxia (3,960 m) and studied by pressure myography. Pregnancy increased UA diameter similarly under normoxia and hypoxia. Although chronic hypoxia raised resting tone in UA from nonpregnant guinea pigs to approximately 20% and tone was greater in preconstricted pregnant chronically hypoxic vs. normoxic UA (both P<0.01), there was an absence of myogenic response (i.e., an increase in tone with rising pressure) in all groups. Pregnancy increased UA distensibility 1.5-fold but did not change stiffness or the stress strain relationship. Compared with vessels from normoxic pregnant animals, hypoxic pregnancy raised UA distensibility fourfold, decreased stiffness (rate constant b=3.80+/-1.06 vs. 8.92+/-1.25, respectively, P<0.01), lowered elastin by 50%, and shifted the stress-strain relationship upward such that four times as much strain was present at a given stress. We concluded that increased distensibility and low myogenic tone contribute to enlarging UA diameter and raising UA blood flow during pregnancy. Chronic hypoxia exaggerates the rise in distensibility and alters the stress-strain relationship in ways that may provoke vascular injury. PMID- 16714415 TI - Glucose kinetics differ between women and men, during and after exercise. AB - As exercise can improve the regulation of glucose and carbohydrate metabolism, it is important to establish biological factors, such as sex, that may influence these outcomes. Glucose kinetics, therefore, were compared between women and men at rest, during exercise, and postexercise. It was hypothesized that glucose flux would be significantly lower in women than men during both the exercise and postexercise periods. Subjects included normal weight, healthy, eumenorrehic women and men, matched for habitual activity level and maximal oxygen uptake per kilogram lean body mass. Testing occurred following 3 days of diet control, with no exercise the day before. Subjects were tested in the overnight-fasted condition with women studied in the midluteal phase of the menstrual cycle. Resting (120 min), exercise (85% lactate threshold, 90 min), and postexercise (180 min) measurements of glucose flux and substrate metabolism were made. During exercise, women had a significantly lower rate of glucose appearance (Ra) (P<0.001) and disappearance (Rd) (P<0.002) compared with men. Maximal values were achieved at 90 min of exercise for both glucose Ra (mean+/-SE: 22.8+/-1.12 micromol.kg body wt-1.min-1 women and 33.6+/-1.79 micromol.kg body wt-1.min-1 men) and glucose Rd (23.2+/-1.26 and 34.1+/-1.71 micromol.kg body wt-1.min-1, respectively). Exercise epinephrine concentration was significantly lower in women compared with men (P<0.02), as was the increment in glucagon from rest to exercise (P<0.04). During the postexercise period, glucose Ra and Rd were also significantly lower in women vs. men (P<0.001), with differences diminishing over time. In conclusion, circulating blood glucose flux was significantly lower during 90 min of moderate exercise, and immediately postexercise, in women compared with men. Sex differences in the glucagon increase to exercise, and/or the epinephrine levels during exercise, may play a role in determining these sex differences in exercise glucose turnover. PMID- 16714416 TI - Respiratory responses to cold water immersion: neural pathways, interactions, and clinical consequences awake and asleep. AB - The ventilatory responses to immersion and changes in temperature are reviewed. A fall in skin temperature elicits a powerful cardiorespiratory response, termed "cold shock," comprising an initial gasp, hypertension, and hyperventilation despite a profound hypocapnia. The physiology and neural pathways of this are examined with data from original studies. The respiratory responses to skin cooling override both conscious and other autonomic respiratory controls and may act as a precursor to drowning. There is emerging evidence that the combination of the reestablishment of respiratory rhythm following apnea, hypoxemia, and coincident sympathetic nervous and cyclic vagal stimulation appears to be an arrhythmogenic trigger. The potential clinical implications of this during wakefulness and sleep are discussed in relation to sudden death during immersion, underwater birth, and sleep apnea. A drop in deep body temperature leads to a slowing of respiration, which is more profound than the reduced metabolic demand seen with hypothermia, leading to hypercapnia and hypoxia. The control of respiration is abnormal during hypothermia, and correction of the hypoxia by inhalation of oxygen may lead to a further depression of ventilation and even respiratory arrest. The immediate care of patients with hypothermia needs to take these factors into account to maximize the chances of a favorable outcome for the rescued casualty. PMID- 16714417 TI - In health and in a normoxic environment, VO2 max is/is not limited primarily by cardiac output and locomotor muscle blood flow. PMID- 16714418 TI - Lactic acid accumulation is an advantage/disadvantage during muscle activity. PMID- 16714420 TI - Mammalian skeletal muscle can convert lactate to glycogen. PMID- 16714421 TI - Presence of SERCA and calcineurin during fetal development of porcine skeletal muscle. AB - Mechanisms involved in skeletal myofiber differentiation during fetal development of large animals are poorly understood. Studies in small animals suggest that the calcineurin (Cn) pathway is involved in myofiber differentiation. Neural activity is a prerequisite for Cn activity, implying maintenance of sustained low intracellular Ca(2+) concentrations. To study the role of Cn in fetal myofiber differentiation, we monitored the temporal and spatial distribution of Cn subunits, sarcoplasmic reticulum Ca(2+) ATPase (SERCA), phospholamban (PLB), and myosin heavy chain (MyHC) isoforms in relation to ingrowing nerves in porcine semitendinosus muscle (m. semitendinosus) at 55 and 75 days of gestation (dg) and at term. Immunofluorescence analysis revealed the presence of Cn subunits and SERCA isoforms at all analyzed stages. Cn distribution was not fiber-type specific, but expression became more prominent at term. At 75 dg, differential SERCA2 expression was accompanied by perinuclear PLB in primary fibers. SERCA1 was expressed in all fiber types at all stages. No specific MyHC isoform distribution was seen in relation to neuromuscular contacts, although neuromuscular contacts were present. From these results we speculate that in porcine m. semitendinosus differential SERCA2 expression precedes differential Cn expression. The question whether the Cn pathway is involved in prenatal myofiber differentiation needs further studies. PMID- 16714422 TI - Laminin-311 (Laminin-6) fiber assembly by type I-like alveolar cells. AB - Two epithelial cell types cover the alveolar surface of the lung. Type II alveolar epithelial cells produce surfactant and, during development or following wounding, give rise to type I cells that are involved in gas exchange and alveolar fluid homeostasis. In culture, freshly isolated alveolar type II cells assume a more squamous (type I-like) appearance within 4 days after plating. They assemble numerous focal adhesions that associate with the actin cytoskeleton at the cell margins. These alveolar epithelial cells lose expression of type II cell markers including SP-C and after 4 days in culture express the type I cell marker T1alpha. Those cells that express T1alpha also deposit fibers of laminin-311 in their matrix. The latter appears to be related to their development of a type I phenotype because freshly isolated, primary type I cells also assemble laminin 311-rich fibers in vitro. A beta1 integrin antibody antagonist inhibits the assembly of laminin-311 matrix fibers. Moreover, the formation of laminin fibers is dependent on the activity of the small GTPases and is perturbed by ML-7, a myosin light chain kinase inhibitor. In summary, our data indicate that assembly of laminin-311 fibers by lung epithelial cells is integrin and actin cytoskeleton dependent, and that these fibers are characteristic of type I alveolar cells. PMID- 16714423 TI - Comparison of processing and sectioning methodologies for arteries containing metallic stents. AB - The histological study of arteries with implanted metallic scaffolding devices, known as stents, remains a technical challenge. Given that the arterial response to stent implantation can sometimes lead to adverse outcomes, including the re accumulation of tissue mass within the stent (or in-stent restenosis), overcoming these technical challenges is a priority for the advancement of research and development in this important clinical field. Essentially, the task is to section the stent-tissue interface with the least amount of disruption of tissue and cellular morphology. Although many methacrylate resin methodologies are successfully applied toward the study of endovascular stents by a variety of research laboratories, the exact formulations, as well as subsequent processing and sectioning methodology, remain largely coveted. In this paper, we describe in detail a methyl methacrylate resin-embedding methodology that can successfully be applied to tungsten carbide blade, as well as saw and grinding sectioning methods and transmission electron microscopy. In addition, we present a comparison of the two sectioning methodologies in terms of their effectiveness with regard to morphological, histochemical, and immunohistochemical analyses. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials. PMID- 16714424 TI - Method of specific detection of apoptosis using formamide-induced DNA denaturation assay. AB - We compared the reliability between apoptosis detection methods, namely, the terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) method and formamide-induced DNA denaturation assay using a monoclonal antibody (MAb) to single-stranded DNA (ssDNA) (formamide-MAb assay). Reaction targets in these methods are different: the TUNEL method recognizes free 3'-OH DNA ends, whereas the formamide-MAb assay detects ssDNA itself (25-30 bp). We found that the formamide-MAb assay immunohistochemically detected apoptotic cells, whereas the TUNEL method detected apoptotic cells as well as mitotic and necrotic cells. The TUNEL method recognized not only 3'-OH DNA ends cleaved by DNase during apoptosis but also constitutive physiological nicking that occurs in DNA duplication and histone posttranslational modifications during mitosis and random DNA breaks during necrotic execution. By electron microscopy, the mean labeling density (the number of 3'-OH DNA ends/nuclear area) obtained by the TUNEL method was determined to be consistently higher than that (the number of ssDNAs/nuclear area) obtained by the formamide-MAb assay. On the basis of these findings, we conclude that the formamide-MAb assay was more specific than the TUNEL method for the detection of apoptotic cells using electron microscopy; however, the labeling intensity of the formamide-MAb assay was slightly weaker than that of the TUNEL method. PMID- 16714426 TI - Flu pandemic. PMID- 16714425 TI - Valsartan, blood pressure reduction, and C-reactive protein: primary report of the Val-MARC trial. AB - Increased levels of high-sensitivity C-reactive protein (hsCRP) are associated with incident hypertension as well as cardiovascular events, and angiotensin II is a potent proinflammatory mediator. However, whether angiotensin receptor blockade lowers hsCRP is uncertain. We performed a randomized trial in which 1668 patients with stage 2 hypertension were treated with 160 mg valsartan or 160/12.5 mg valsartan/hydrochlorothiazide (HCTZ) once daily for 2 weeks with forced titration to 320 mg valsartan or 320/12.5 mg valsartan/HCTZ for an additional 4 weeks. After 6 weeks, systolic blood pressure (-25 versus -18 mm Hg; P<0.001) and diastolic blood pressure (-14 versus -9 mm Hg; P<0.001) were reduced to a greater degree among those allocated to valsartan/HCTZ than to valsartan monotherapy. The median change in hsCRP was -0.12 mg/L among those allocated to valsartan compared with +0.05 mg/L among those allocated to valsartan/HCTZ, a 13.3% difference (P<0.001); this difference between valsartan and valsartan/HCTZ was present in all subgroups evaluated despite the fact that blood pressure reduction was greater in the combined therapy group. No relationship was observed between hsCRP reduction and blood pressure; in all analyses, the proportion of variation in change in hsCRP with valsartan monotherapy explained by change in blood pressure was <2%. Thus, in this prospective trial, valsartan reduced hsCRP levels in a manner independent of degree of blood pressure reduction. These data raise the hypothesis that angiotensin receptor blockade may have anti-inflammatory effects in addition to blood pressure-lowering effects. PMID- 16714427 TI - Noncanonical RNAs from transcripts of the Drosophila muscleblind gene. AB - It has become increasingly evident that eukaryotic cells produce RNA molecules from coding genes with constitutions other than those of typically spliced mRNA transcripts. Here we describe new cDNAs from the Drosophila melanogaster muscleblind (mbl) locus that identify two such atypical RNA molecules: RNAs containing an incomplete exon 2 tandem repetition (mblE2E2') or having exons with a different order compared to the corresponding genomic DNA (mblE2E3'E2'; exon scrambling). The existence of exon duplications and rearrangements in the genomic locus that might explain such cDNAs was ruled out by genomic Southern blotting and in silico analysis of the Drosophila genome sequence. The incomplete exon 2 tandem repetition was confirmed by sequencing reverse transcriptase-polymerase chain reaction (RT-PCR) products, rapid amplification of cDNA ends, and detection of a band consistent with cDNA sizes in total RNA northern blots. RT-PCRs with exon-specific primers downstream of exon 2 were unable to amplify products other than those expected from canonical mbl isoforms, thus indicating that no other exons were efficiently spliced downstream of exon 2. Moreover, mblE2E2' transcripts seem to be poorly polyadenylated, if at all, and behave aberrantly in a polyacrylamide gel electrophoresis (PAGE) mobility assay. Taken together, lack of polyadenylation, lack of downstream splicing events, small size of mblE2E2', and PAGE behavior all suggest that these noncanonical transcripts may be circular RNAs. The functional implications for these noncanonical transcripts are unclear. A developmental expression profile of mblE2E2' revealed an almost constant expression except during early embryogenesis and early adulthood. The protein putatively encoded is unlikely to be functional because an in-frame stop codon occurs almost immediately after the splice site. Such noncanonical transcripts have previously been observed in vertebrates, and these data provide the first experimental evidence for similar phenomena in invertebrates. PMID- 16714428 TI - Unusual intraindividual variation of the nuclear 18S rRNA gene is widespread within the Acipenseridae. AB - Significant intraindividual variation in the sequence of the 18S rRNA gene is unusual in animal genomes. In a previous study, multiple 18S rRNA gene sequences were observed within individuals of eight species of sturgeon from North America but not in the North American paddlefish, Polyodon spathula, in two species of Polypterus (Polypterus delhezi and Polypterus senegalus), in other primitive fishes (Erpetoichthys calabaricus, Lepisosteus osseus, Amia calva) or in a lungfish (Protopterus sp.). These observations led to the hypothesis that this unusual genetic characteristic arose within the Acipenseriformes after the presumed divergence of the sturgeon and paddlefish families. In the present study, a survey of nearly all Eurasian acipenseriform species was conducted to examine 18S rDNA variation. Intraindividual variation was not found in the polyodontid species, the Chinese paddlefish, Psephurus gladius, but variation was detected in all Eurasian acipenserid species. The comparison of sequences from two major segments of the 18S rRNA gene and identification of sites where insertion/deletion events have occurred are placed in the context of evolutionary relationships within the Acipenseriformes and the evolution of rDNA variation in this group. PMID- 16714430 TI - Measurements of footpad dermatitis in broiler chickens at processing plants. AB - A four-point photographic scale was developed to score the severity of lesions of footpad dermatitis (FPD) in broiler chickens. There was a linear relationship between the square root of the relative area of the lesions and their scores, and good agreement between different assessors using the scale. The scale was used to assess samples of 100 birds from each of 190 flocks slaughtered at two uk processing plants in 2002 and 2003; 12 of the flocks (6.3 per cent) had no lesions, but the others had lesions of different prevalences and severity. The maximum proportion of affected birds in a flock was 92 per cent. In the 178 affected flocks, 84.0 per cent of the birds had no lesions and 16.0 per cent had some evidence of FPD. The overall unweighted prevalence of birds with FPD in all 190 flocks sampled was 18.1 per cent; 10.2 per cent had only small lesions, on average equivalent in area to 2.1 per cent of the total area of the foot, 6.2 per cent had lesions on average equivalent to 6.6 per cent of the area of the foot, and 1.7 per cent had lesions on average equivalent to 21.5 per cent of the area of the foot. There were differences between the two plants in the overall prevalence and severity of FPD, but this may have been due to the fact that the plants were sampled at different times of the year. PMID- 16714431 TI - Estimates of the prevalence of transmissible spongiform encephalopathies in sheep and goats in France in 2002. AB - An active surveillance programme for transmissible spongiform encephalopathies (TSES) in sheep and goats was implemented in France in 2002 at abattoirs and rendering plants. The analysis of the results of this programme highlighted three biases: a potentially non-random sampling scheme in both rendering plants and abattoirs, a heterogeneous geographical sampling ratio, and the use of two diagnostic tests of unequal sensitivity. Simulations were run to estimate the prevalence of TSES by taking these biases into account. A comparison of the prevalence of TSES calculated from the raw data with the simulation results showed that the effects of non-random sampling were minor in comparison with the effects of the heterogeneous geographical sampling ratio and the use of two diagnostic tests. PMID- 16714432 TI - Characteristics, prevalence and aetiology of lesions of the oral mucosa in adult sheep. AB - A total of 214 sheep with lesions of the oral mucosa were recorded at the Veterinary Laboratories Agency regional laboratories between March 15, 2002 and February 28, 2003. Using denominator data from the scrapie abattoir survey a prevalence of 0.95 per cent (95 per cent CI 0.82 to 1.10 per cent) was recorded, and using the fallen stock survey a prevalence of 1.15 per cent (95 per cent CI 0.64 to 1.91 per cent) was recorded. The lesions varied widely and included dental pad lesions in 20 per cent of cases. In total, 248 sites were affected, the commonest being the lower gum below the incisors, which was affected in 100 sheep. Most of the lesions were 1 cm or less in diameter. Ninety per cent of the 251 lesions examined histologically were erosions, ulcers, healed ulcers, focal epithelial necrosis or haemorrhages, and the changes observed indicated that trauma was the most likely primary cause. PMID- 16714433 TI - Pathological changes in the bone marrow of dogs with leishmaniosis. AB - Bone marrow aspiration smears from 15 dogs naturally infected with leishmania were evaluated. Three of the dogs showed no clinical signs, six had up to three clinical signs and six had more than three. The most common pathological features of the bone marrow were megakaryocytic dysplasia in 10 of the dogs, erythrophagocytosis in eight, erythroid dysplasia in two and emperipolesis in two. The megakaryocytic and erythroid dysplasia were probably related to an increased number of marrow macrophages producing high levels of tumour necrosis factor alpha and interferon gamma. Six of the dogs with clinical signs showed bone marrow dysplastic features and erythrophagocytosis, suggesting that leishmaniosis could be the unique cause of both conditions. PMID- 16714434 TI - Risk assessments to inform policy decisions regarding importation of pets from North America. PMID- 16714435 TI - Spinal tumours in three cows. PMID- 16714436 TI - Ileocolic intussusception in a pine snake (Pituophis melanoleucus). PMID- 16714437 TI - Prevalence of Brachyspira species in pigs with diarrhoea in Spain. PMID- 16714438 TI - Acute basophilic leukaemia in a calf. PMID- 16714439 TI - FMD control strategies. PMID- 16714440 TI - Update on low pathogenic avian influenza. PMID- 16714443 TI - mazF, a novel counter-selectable marker for unmarked chromosomal manipulation in Bacillus subtilis. AB - Here, we present a novel method for the directed genetic manipulation of the Bacillus subtilis chromosome free of any selection marker. Our new approach employed the Escherichia coli toxin gene mazF as a counter-selectable marker. The mazF gene was placed under the control of an isopropyl-beta-D thiogalactopyranoside (IPTG)-inducible expression system and associated with a spectomycin-resistance gene to form the MazF cassette, which was flanked by two directly-repeated (DR) sequences. A double-crossover event between the linearized delivery vector and the chromosome integrated the MazF cassette into a target locus and yielded an IPTG-sensitive strain with spectomycin-resistance, in which the wild-type chromosome copy had been replaced by the modified copy at the targeted locus. Another single-crossover event between the two DR sequences led to the excision of the MazF cassette and generated a strain with IPTG resistance, thereby realizing the desired alteration to the chromosome without introducing any unwanted selection markers. We used this method repeatedly and successfully to inactivate a specific gene, to introduce a gene of interest and to realize the in-frame deletion of a target gene in the same strain. As there is no prerequisite strain for this method, it will be a powerful and universal tool. PMID- 16714444 TI - The tale beyond the tail: histone core domain modifications and the regulation of chromatin structure. AB - Histone post-translational modifications occur, not only in the N-terminal tail domains, but also in the core domains. While modifications in the N-terminal tail function largely through the regulation of the binding of non-histone proteins to chromatin, based on their location in the nucleosome, core domain modifications may also function through distinct mechanisms involving structural alterations to the nucleosome. This article reviews the recent developments in regards to these novel histone modifications and discusses their important role in the regulation of chromatin structure. PMID- 16714446 TI - Evolution of small nucleolar RNAs in nematodes. AB - In contrast to mRNAs, which are templates for translating proteins, non-protein coding (npc) RNAs (also known as 'non-coding' RNA, ncRNA), exhibit various functions in different compartments and developmental stages of the cell. Small nucleolar RNAs (snoRNAs), one of the largest classes of npcRNAs, guide post transcriptional modifications of other RNAs that are crucial for appropriate RNA folding as well as for RNA-RNA and RNA-protein interactions. Although snoRNA genes comprise a significant fraction of the eutherian genome, identifying and characterizing large numbers of them is not sufficiently accessible by classical computer searches alone. Furthermore, most previous investigations of snoRNAs yielded only limited indications of their evolution. Using data obtained by a combination of high-throughput cDNA library screening and computational search strategies based on a modified DNAMAN program, we characterized 151 npcRNAs, and in particular 121 snoRNAs, from Caenorhabditis elegans and extensively compared them with those in the related, Caenorhabditis briggsae. Detailed comparisons of paralog snoRNAs in the two nematodes revealed, in addition to trans-duplication, a novel, cis-duplication distribution strategy with insertions near to the original loci. Some snoRNAs coevolved with their modification target sites, demonstrating the close interaction of complementary regions. Some target sites modified by snoRNAs were changed, added or lost, documenting a high degree of evolutionary plasticity of npcRNAs. PMID- 16714445 TI - Long homopurine*homopyrimidine sequences are characteristic of genes expressed in brain and the pseudoautosomal region. AB - Homo(purine*pyrimidine) sequences (R*Y tracts) with mirror repeat symmetries form stable triplexes that block replication and transcription and promote genetic rearrangements. A systematic search was conducted to map the location of the longest R*Y tracts in the human genome in order to assess their potential function(s). The 814 R*Y tracts with > or =250 uninterrupted base pairs were preferentially clustered in the pseudoautosomal region of the sex chromosomes and located in the introns of 228 annotated genes whose protein products were associated with functions at the cell membrane. These genes were highly expressed in the brain and particularly in genes associated with susceptibility to mental disorders, such as schizophrenia. The set of 1957 genes harboring the 2886 R*Y tracts with > or =100 uninterrupted base pairs was additionally enriched in proteins associated with phosphorylation, signal transduction, development and morphogenesis. Comparisons of the > or =250 bp R*Y tracts in the mouse and chimpanzee genomes indicated that these sequences have mutated faster than the surrounding regions and are longer in humans than in chimpanzees. These results support a role for long R*Y tracts in promoting recombination and genome diversity during evolution through destabilization of chromosomal DNA, thereby inducing repair and mutation. PMID- 16714447 TI - Affinity of molecular interactions in the bacteriophage phi29 DNA packaging motor. AB - DNA packaging in the bacteriophage phi29 involves a molecular motor with protein and RNA components, including interactions between the viral connector protein and molecules of pRNA, both of which form multimeric complexes. Data are presented to demonstrate the higher order assembly of pRNA together with the affinity of pRNA:pRNA and pRNA:connector interactions, which are used to propose a model for motor function. In solution, pRNA can form dimeric and trimeric multimers in a magnesium-dependent manner, with dissociation constants for multimerization in the micromolar range. pRNA:connector binding is also facilitated by the presence of magnesium ions, with a nanomolar apparent dissociation constant for the interaction. From studies with a mutant pRNA, it appears that multimerization of pRNA is not essential for connector binding and it is likely that connector protein is involved in the stabilization of higher order RNA multimers. It is proposed that magnesium ions may promote conformational change that facilitate pRNA:connector interactions, essential for motor function. PMID- 16714448 TI - Modulation of telomere terminal structure by telomerase components in Candida albicans. AB - The telomerase ribonucleoprotein in Candida albicans is presumed to contain at least three Est proteins: CaEst1p, CaEst2p/TERT and CaEst3p. We constructed mutants missing each of the protein subunit of telomerase and analyzed overall telomere dynamics and single-stranded telomere overhangs over the course of many generations. The est1-DeltaDelta mutant manifested abrupt telomere loss and recovery, consistent with heightened recombination. Both the est2-DeltaDelta and est3-DeltaDelta mutant exhibited progressive telomere loss, followed by the gradual emergence of survivors with long telomeres. In no case was telomere loss accompanied by severe growth defects, suggesting that cells with short telomeres can continue to proliferate. Furthermore, the amount of G-strand terminal overhangs was greatly increased in the est2-DeltaDelta mutant, but not others. Our results suggest that in addition to their well-characterized function in telomere elongation, both CaEst1p and CaEst2p mediate some aspects of telomere protection in Candida, with the former suppressing excessive recombination, and the latter preventing excessive C-strand degradation. PMID- 16714449 TI - Human telomeric sequence forms a hybrid-type intramolecular G-quadruplex structure with mixed parallel/antiparallel strands in potassium solution. AB - Human telomeric DNA consists of tandem repeats of the sequence d(TTAGGG). The formation and stabilization of DNA G-quadruplexes in the human telomeric sequence have been shown to inhibit the activity of telomerase, thus the telomeric DNA G quadruplex has been considered as an attractive target for cancer therapeutic intervention. However, knowledge of the intact human telomeric G-quadruplex structure(s) formed under physiological conditions is a prerequisite for structure-based rational drug design. Here we report the folding structure of the human telomeric sequence in K+ solution determined by NMR. Our results demonstrate a novel, unprecedented intramolecular G-quadruplex folding topology with hybrid-type mixed parallel/antiparallel G-strands. This telomeric G quadruplex structure contains three G-tetrads with mixed G-arrangements, which are connected consecutively with a double-chain-reversal side loop and two lateral loops, each consisting of three nucleotides TTA. This intramolecular hybrid-type telomeric G-quadruplex structure formed in K+ solution is distinct from those reported on the 22 nt Tel22 in Na+ solution and in crystalline state in the presence of K+, and appears to be the predominant conformation for the extended 26 nt telomeric sequence Tel26 in the presence of K+, regardless of the presence or absence of Na+. Furthermore, the addition of K+ readily converts the Na+-form conformation to the K+-form hybrid-type G-quadruplex. Our results explain all the reported experimental data on the human telomeric G-quadruplexes formed in the presence of K+, and provide important insights for understanding the polymorphism and interconversion of various G-quadruplex structures formed within the human telomeric sequence, as well as the effects of sequence and cations. This hybrid-type G-quadruplex topology suggests a straightforward pathway for the secondary structure formation with effective packing within the extended human telomeric DNA. The hybrid-type telomeric G-quadruplex is most likely to be of pharmacological relevance, and the distinct folding topology of this G-quadruplex suggests that it can be specifically targeted by G-quadruplex interactive small molecule drugs. PMID- 16714451 TI - Science to practice: what is new about detecting emphysema? AB - Through the use of diffusion-weighted 3He MR imaging, Fain et al in this issue of Radiology demonstrated the potential for detection of early emphysema in the lungs of healthy smokers. The technique possibly could become an important tool for understanding the pulmonary processes that occur with emphysema, for implementing treatment before clinical symptoms occur, and for monitoring the effect of treatment. PMID- 16714452 TI - Education and cultural development of the health care work force. Part I. The health professions. PMID- 16714450 TI - Collaboration of Werner syndrome protein and BRCA1 in cellular responses to DNA interstrand cross-links. AB - Cells deficient in the Werner syndrome protein (WRN) or BRCA1 are hypersensitive to DNA interstrand cross-links (ICLs), whose repair requires nucleotide excision repair (NER) and homologous recombination (HR). However, the roles of WRN and BRCA1 in the repair of DNA ICLs are not understood and the molecular mechanisms of ICL repair at the processing stage have not yet been established. This study demonstrates that WRN helicase activity, but not exonuclease activity, is required to process DNA ICLs in cells and that WRN cooperates with BRCA1 in the cellular response to DNA ICLs. BRCA1 interacts directly with WRN and stimulates WRN helicase and exonuclease activities in vitro. The interaction between WRN and BRCA1 increases in cells treated with DNA cross-linking agents. WRN binding to BRCA1 was mapped to BRCA1 452-1079 amino acids. The BRCA1/BARD1 complex also associates with WRN in vivo and stimulates WRN helicase activity on forked and Holliday junction substrates. These findings suggest that WRN and BRCA1 act in a coordinated manner to facilitate repair of DNA ICLs. PMID- 16714453 TI - Financial relationships with industry and device research involving non-Food and Drug Administration-approved use: a perspective. PMID- 16714454 TI - Bridging the knowledge divide in radiology education. PMID- 16714455 TI - Diffusion-weighted and perfusion MR imaging for brain tumor characterization and assessment of treatment response. AB - Diffusion-weighted magnetic resonance (MR) imaging and perfusion MR imaging are advanced techniques that provide information not available from conventional MR imaging. In particular, these techniques have a number of applications with regard to characterization of tumors and assessment of tumor response to therapy. In this review, the authors describe the fundamental principles of diffusion weighted and perfusion MR imaging and provide an overview of the ways in which these techniques are being used to characterize tumors by helping distinguish tumor types, assess tumor grade, and attempt to determine tumor margins. In addition, the role of these techniques for evaluating response to tumor therapy is outlined. PMID- 16714456 TI - Intracranial cysts: radiologic-pathologic correlation and imaging approach. AB - Cysts and cystic-appearing intracranial masses have a broad imaging and pathologic spectra. The authors review the pathologic findings, origin, radiologic appearance, and differential diagnosis of many different intracranial cysts. A diagnostic algorithm based on most common anatomic locations is presented that helps narrow the differential diagnosis. PMID- 16714457 TI - Evaluation of a prospective scoring system designed for a multicenter breast MR imaging screening study. AB - PURPOSE: To evaluate prospectively the accuracy of a lesion classification system designed for use in a magnetic resonance (MR) imaging high-breast-cancer-risk screening study. MATERIALS AND METHODS: All participating patients provided written informed consent. Ethics committee approval was obtained. The results of 1541 contrast material-enhanced breast MR imaging examinations were analyzed; 1441 screening examinations were performed in 638 women aged 24-51 years at high risk for breast cancer, and 100 examinations were performed in 100 women aged 23 81 years. Lesion analysis was performed in 991 breasts, which were divided into design (491 breasts) and testing (500 breasts) sets. The reference standard was histologic analysis of biopsy samples, fine-needle aspiration cytology, or minimal follow-up of 24 months. The scoring system involved the use of five features: morphology (MOR), pattern of enhancement (POE), percentage of maximal focal enhancement (PMFE), maximal signal intensity-time ratio (MITR), and pattern of contrast material washout (POCW). The system was evaluated by means of (a) assessment of interreader agreement, as expressed in kappa statistics, for 315 breasts in which both readers analyzed the same lesion, (b) assessment of the diagnostic accuracy of the scored components with receiver operating characteristic curve analysis, and (c) logistic regression analysis to determine which components of the scoring system were critical to the final score. A new simplified scoring system developed with the design set was applied to the testing set. RESULTS: There was moderate reader agreement regarding overall lesion outcome (ie, malignant, suspicious, or benign) (kappa=0.58) and less agreement regarding the scored components. The area under the receiver operating characteristic curve (AUC) for the overall lesion score, 0.88, was higher than the AUC for any one component. The components MOR, POE, and POCW yielded the best overall result. PMFE and MITR did not contribute to diagnostic utility. Applying a simplified scoring system to the testing set yielded a nonsignificantly (P=.2) higher AUC than did applying the original scoring system (sensitivity, 84%; specificity, 86.0%). CONCLUSION: Good diagnostic accuracy can be achieved by using simple qualitative descriptors of lesion enhancement, including POCW. In the context of screening, quantitative enhancement parameters appear to be less useful for lesion characterization. PMID- 16714458 TI - Angiogenesis: noninvasive quantitative assessment with contrast-enhanced functional US in murine model. AB - PURPOSE: To evaluate quantitative functional ultrasonography (US) in a murine gel model by using microbubble destruction kinetics to determine whether parametric indices provided with US could help assess angiogenesis. MATERIALS AND METHODS: Institutional Animal Subjects Committee approved experiments and procedures. In 36 normal mice, two 0.4-mL gel implants were placed subcutaneously on either side of spine. One implant contained 0.5, 1.0, or 1.5 microg human basic fibroblast growth factor (bFGF) per milliliter of gel. Functional US quantitative analysis of angiogenesis with microbubble contrast agent was performed on days 3, 6, 9, and 12; histologic data were collected. Time-intensity curve of implant was fitted to mathematic decay model to calculate fractional blood volume and fraction of blood replaced per unit of time. Microvascular density (MVD) and percentage of microvascular area (MVA) were measured after anti-CD31 staining. Spearman rank order correlation was used in analyses. RESULTS: bFGF-containing implants induced MVD of eight, 35, 42, and 42 vessels per square millimeter on days 3, 6, 9, and 12, respectively; in controls, MVD was four vessels/mm2 (P<.05 on days 6, 9, and 12). bFGF-containing implants induced percentage MVA of 2%, 5%, 20%, and 27%, respectively; in controls, it was 0.5% (P<.05). Maximum enhancement was significantly increased in bFGF implants (23.3 gray level+/-14.1 [standard deviation]) compared with controls (11.0+/-5.5, P<.001). Implants containing bFGF showed poor correlations between fractional blood volume and MVD (r2=0.42) or percentage MVA (r2=0.51) at US. There was no correlation between microbubble velocity and MVD (r2<0.05) or percentage MVA (r2<0.13). CONCLUSION: Functional US perfusion parameters do not correlate with current histologic indices for quantifying angiogenesis. MVD, as a histologic quantitative measurement of angiogenesis, may not be an appropriate standard for contrast-enhanced imaging that relies on perfused neovessels. PMID- 16714459 TI - Internal barium shielding to minimize fetal irradiation in spiral chest CT: a phantom simulation experiment. AB - PURPOSE: To use a phantom to prospectively examine the attenuating effect of barium sulfate as an internal shield to protect the fetus. MATERIALS AND METHODS: In an adult-size phantom, 1- and 2-cm-thick acrylic slabs containing 315 or 630 mL of water, 2% or 40% barium sulfate suspension, and a 1-mm lead sheet were placed under the diaphragm. In 17 experiments, fetal dose was measured by using thermoluminescent dosimeters that were placed immediately under (near field) and 10 cm below (far field) the water slab (eight experiments), barium sulfate slab (eight experiments), and lead sheet (one experiment). In a pulmonary embolism protocol, the phantom was scanned with single-detector spiral computed tomography (CT) at 130 kVp and 230 mAs. RESULTS: The control radiation dose was 3.60 mSv+/ 0.54 (standard deviation) with the water slab at near field, where the uterus dome is at near term, and 0.507 mSv+/-0.07 with the water slab at far field, the uterus position during early gestation. Scattered radiation was attenuated 13% and 21% with 2% barium sulfate and 87% and 96% with 40% barium sulfate, as calculated in the near and far fields, respectively, and 99% with the 1-mm lead sheet. The extrapolated attenuations for 5%-40% barium sulfate suspensions indicated that beyond a 30% suspension, attenuation increased further only slightly. CONCLUSION: Study results in the phantom experiment suggest that fetal irradiation during maternal chest CT can be reduced substantially with barium shielding. PMID- 16714460 TI - CT colonography: influence of 3D viewing and polyp candidate features on interpretation with computer-aided detection. AB - PURPOSE: To retrospectively determine if three-dimensional (3D) viewing improves radiologists' accuracy in classifying true-positive (TP) and false-positive (FP) polyp candidates identified with computer-aided detection (CAD) and to determine candidate polyp features that are associated with classification accuracy, with known polyps serving as the reference standard. MATERIALS AND METHODS: Institutional review board approval and informed consent were obtained; this study was HIPAA compliant. Forty-seven computed tomographic (CT) colonography data sets were obtained in 26 men and 10 women (age range, 42-76 years). Four radiologists classified 705 polyp candidates (53 TP candidates, 652 FP candidates) identified with CAD; initially, only two-dimensional images were used, but these were later supplemented with 3D rendering. Another radiologist unblinded to colonoscopy findings characterized the features of each candidate, assessed colon distention and preparation, and defined the true nature of FP candidates. Receiver operating characteristic curves were used to compare readers' performance, and repeated-measures analysis of variance was used to test features that affect interpretation. RESULTS: Use of 3D viewing improved classification accuracy for three readers and increased the area under the receiver operating characteristic curve to 0.96-0.97 (P<.001). For TP candidates, maximum polyp width (P=.038), polyp height (P=.019), and preparation (P=.004) significantly affected accuracy. For FP candidates, colonic segment (P=.007), attenuation (P<.001), surface smoothness (P<.001), distention (P=.034), preparation (P<.001), and true nature of candidate lesions (P<.001) significantly affected accuracy. CONCLUSION: Use of 3D viewing increases reader accuracy in the classification of polyp candidates identified with CAD. Polyp size and examination quality are significantly associated with accuracy. PMID- 16714462 TI - Lateral meniscus root tear and meniscus extrusion with anterior cruciate ligament tear. AB - PURPOSE: To retrospectively determine the prevalence of posterior lateral meniscus root tears (LMRTs), as depicted on magnetic resonance (MR) images, in patients with an anterior cruciate ligament (ACL) tear and to investigate the association of LMRTs with lateral meniscus extrusion and other ligament injuries. MATERIALS AND METHODS: Institutional review board approval was obtained; informed consent was not required. This study was HIPAA compliant. MR images were obtained in 174 male and 119 female patients (mean age, 37 years; age range, 16-87 years) and retrospectively reviewed for LMRT, medial meniscus root tear (MMRT), nonroot meniscus tear, meniscus extrusion, and presence of meniscofemoral ligaments (MFLs). The chi2 and unpaired Student t tests were performed. RESULTS: In 33 patients, 34 meniscus root tears were identified. An LMRT was present in 26 (9.8%) of 264 patients, and an MMRT was present in eight (3.0%) (P=.008). Lateral meniscus extrusion was present in six (23%) of 26 LMRTs and five (2.2%) of 231 patients with normal meniscus roots (P<.001). Complex or deep radial tears were found in three of five cases of lateral meniscus extrusion and normal root. The MFL was not observed in five (19%) of 26 studies of an LMRT. Among these 26 studies of an LMRT, lateral meniscus extrusion was identified in three (14%) of 21 cases in which the MFL was intact and in three (60%) of five cases in which the MFL was not identified (P<.03). Prevalence of an extruded meniscus was seven (88%) of eight for an MMRT and six (23%) of 26 for an LMRT (P=.001). CONCLUSION: Prevalence of LMRTs is greater than that of MMRTs in patients with an ACL tear. LMRTs and complex and radial tears are associated with lateral meniscus extrusion; an absent MFL is more prevalent in patients with LMRTs and when the meniscus is extruded. PMID- 16714461 TI - Prophylaxis of contrast material-induced nephropathy in patients in intensive care: acetylcysteine, theophylline, or both? A randomized study. AB - PURPOSE: To prospectively compare the protective effect of acetylcysteine, theophylline, and both agents combined in patients who are admitted to the intensive care unit with at least one risk factor for contrast material-induced nephropathy and who receive at least 100 mL of iodinated contrast medium. MATERIALS AND METHODS: Institutional ethics review board approval and informed consent were obtained. A total of 91 patients (mean age, 58.5 years+/-14.8 [standard deviation]; 31 women, 60 men; 150 examinations) were admitted to the intensive care unit with at least one risk factor for contrast-induced nephropathy and received either (a) 200 mg theophylline 30 minutes before contrast medium administration (group T), (b) 600 mg acetylcysteine twice daily on the day of and (if possible) the day before the examination (group A), or (c) both agents combined (group AT). The primary endpoint for this study was the incidence of contrast-induced nephropathy (chi2 test). RESULTS: Groups T, A, and AT were comparable with regard to baseline creatinine levels and the amount of contrast medium administered. The incidence of contrast-induced nephropathy in groups T, A, and AT was 2%, 12%, and 4%, respectively, and was significantly lower in group T than in group A (P=.047). There was no significant difference in the incidence of contrast-induced nephropathy between groups A and AT (P=.148) or between groups T and AT (P=.53). For group A, serum creatinine did not change after 12, 24, or 48 hours compared with baseline. Creatinine levels in group T decreased 12 hours (1.19 mg/dL+/-0.58; P=.008) and 48 hours (1.16 mg/dL+/-0.55; P=.034) after contrast material injection compared with baseline (1.25 mg/dL+/ 0.61). In group AT, creatinine significantly decreased 24 hours (1.21 mg/dL+/ 0.74; P=.003) and 48 hours (1.17 mg/dL+/-0.69; P<.001) after contrast material injection compared with baseline (1.28 mg/dL+/-0.74). Group A had significantly higher maximal increases in creatinine than groups T and AT (P=.014). CONCLUSION: For prophylaxis of contrast-induced nephropathy in patients who are admitted to the intensive care unit and who receive 100 mL or more of contrast medium, theophylline is superior to acetylcysteine. PMID- 16714463 TI - Osteoarthritis of the knee: association between clinical features and MR imaging findings. AB - PURPOSE: To prospectively evaluate the association between clinical features and structural abnormalities found at magnetic resonance (MR) imaging in patients with osteoarthritis (OA) of the knee. MATERIALS AND METHODS: The study was approved by the institutional medical ethics review board. Written informed consent was obtained from each patient. MR images of the knee were obtained from 205 (42 [20%] men, 163 [80%] women; median age, 60 years; range, 43-77 years) patients in whom symptomatic OA at multiple joint sites was diagnosed. MR images were analyzed for various abnormalities of OA. All patients were interviewed concerning pain and stiffness in the knee that was imaged. Odds ratios (ORs) with 99% confidence intervals (CIs) were used to determine the association between the imaging findings and clinical features of OA. RESULTS: A large joint effusion was associated with pain (OR, 9.99; 99% CI: 1.28, 149) and stiffness (OR, 4.67; 99% CI: 1.26, 26.1). The presence of an osteophyte in the patellofemoral compartment (OR, 2.25; 99% CI: 1.06, 4.77) was associated with pain. All other imaging findings, including focal or diffuse cartilaginous abnormalities, subchondral cysts, bone marrow edema, subluxation of the meniscus, meniscal tears, or Baker cysts, were not associated with symptoms. CONCLUSION: Findings of this study indicate that only two associations exist between clinical symptoms and structural findings found on MR images in patients with OA of the knee. PMID- 16714464 TI - MR imaging of middle cerebellar peduncle width: differentiation of multiple system atrophy from Parkinson disease. AB - PURPOSE: To prospectively assess if middle cerebellar peduncle (MCP) atrophy, evaluated at magnetic resonance (MR) imaging, can help differentiate multiple system atrophy (MSA) from Parkinson disease (PD). MATERIALS AND METHODS: All participants provided informed consent for participation in the study, which was approved by the institutional review board. Sixteen consecutive patients with MSA, 26 consecutive patients with PD, and 14 healthy control subjects were examined with MR imaging. Images were interpreted independently by two experienced neuroradiologists blinded to clinical information, who visually inspected the images for the presence or absence of putaminal atrophy, putaminal hypointensity, slitlike hyperintensity in the posterolateral margin of the putamen, brainstem atrophy, hyperintensity of the MCP, and cruciform hyperintensity of the pons. Measurements of MCP width on T1-weighted volumetric spoiled gradient-echo images were performed in all subjects. Differences in MCP width among the groups were evaluated by using the Kruskall-Wallis test, followed by the Mann-Whitney U test for multiple comparisons and Bonferroni correction. RESULTS: All patients (mean age, 63.88 years; range, 55-72 years) with MSA had at least one of the features commonly observed in this disease on MR images, whereas control subjects (mean age, 66.93 years; range, 61-77 years) and all but one patient with PD (mean age, 65.31 years; range, 51-79 years) had normal MR images. The average MCP width was significantly smaller in patients with MSA (6.10 mm+/ 1.18 [standard deviation]) than in those with PD (9.32 mm+/-0.77, P<.001) or control subjects (9.80 mm+/-0.66, P<.001). CONCLUSION: Measurement of MCP width on MR images may be useful for distinguishing patients with MSA from those with PD. PMID- 16714467 TI - Case 95: fracture of double-layered patella in multiple epiphyseal dysplasia. PMID- 16714465 TI - Early emphysematous changes in asymptomatic smokers: detection with 3He MR imaging. AB - PURPOSE: To prospectively compare apparent diffusion coefficient (ADC) measurements derived from diffusion-weighted hyperpolarized helium 3 (3He) magnetic resonance (MR) imaging with functional and structural findings using spirometric tests and thin-section computed tomography (CT) of the lungs in asymptomatic smokers and healthy nonsmokers of similar age. MATERIALS AND METHODS: All studies were HIPAA compliant and were approved by the institutional review board. Informed consent was obtained. Ventilation and diffusion-weighted 3He MR images were obtained in healthy subjects: 11 smokers (five women, six men; mean age, 47 years+/-18 [standard deviation]; range, 23-73 years) and eight nonsmokers (<100 cigarettes in lifetime) (four women, four men; mean age, 46 years+/-16; range, 23-69 years). Mean ADC values for smokers and nonsmokers were compared with spirometric values, diffusing capacity of the lung for carbon monoxide (Dlco), age, and pack-years with Spearman rank correlation coefficient (rs) and multiple linear regression analysis. Mean ADC value and thin-section CT emphysema index of relative area less than -950 HU (RA950) were compared on a regional basis by using linear mixed-effect models. RESULTS: Mean ADC values and number of pack-years were significantly correlated (rs=0.60; 95% confidence interval (CI): 0.21, 1.00; P=.007); relationship remained significant after adjustment for age (P=.003). Dlco was strongly correlated with pack-years (rs= 0.63; 95% CI: -0.97, -0.29; P=.004). Negative correlations between mean ADC values and percentage predicted Dlco (rs=-0.79; 95% CI: -0.93, -0.64; P<.001) and the ratio of forced expiratory volume in 1 second to forced vital capacity (rs= 0.72; 95% CI: -0.92, -0.52; P=.001) were statistically significant. Correlations between spirometric values or RA950 and number of pack-years were not significant (.05 level). CONCLUSION: Correlations between mean ADC values and pulmonary function test measurements for diagnosing emphysema, especially Dlco, were statistically significant. PMID- 16714468 TI - The anterior tibial translocation sign. PMID- 16714469 TI - Computer-aided detection with screening mammography: improving performance or simply shifting the operating point? PMID- 16714470 TI - In vitro study of various iodinated contrast media. PMID- 16714472 TI - Pneumocystis pneumonia--Los Angeles. 1981. PMID- 16714473 TI - A failure to communicate . . . PMID- 16714474 TI - The kindling fluorescent protein: a transient photoswitchable marker. AB - Passive fluorescent protein markers are indispensable for dynamic cellular imaging; however, they are unselective, introduce constant background fluorescence, and require continuous observation. Photoactivatable fluorescent proteins have now been developed whose fluorescence can be switched on and off by illumination, allowing selective and direct tracking of tagged objects without the need for continuous imaging. The "kindling fluorescent protein" is a photoactivatable marker with a novel twist: it turns itself off after a selectable period. PMID- 16714471 TI - Factors mediating cognitive deficits and psychopathology among siblings of individuals with schizophrenia. AB - Schizotypal traits and cognitive disturbances are known to be present in first degree relatives of people with schizophrenia. However, there is little understanding of how these endophenotypes are related to each other. We explored the nature of this relationship in individuals with schizophrenia, their full siblings, community controls, and their siblings. All participants were assessed in the domains of working memory, attention, episodic memory, and executive function, as well as in their level of positive, negative, and disorganization symptoms. Schizophrenia probands were significantly impaired on all cognitive domains, as compared with the other 3 groups, and displayed the highest levels of positive, negative, and disorganization symptoms. Proband siblings performed significantly worse than controls on tasks of working memory, episodic memory, and executive function, and they displayed significantly more positive and negative symptoms as compared with controls. Poorer task performance across all 4 cognitive domains was most strongly correlated with increased negative symptoms. Mediation analyses revealed that working memory, episodic memory, and executive function deficits partially mediated increases in negative symptoms among proband siblings. Negative symptoms fully mediated deficits in working memory and episodic memory but only partially mediated deficits in executive function. Results suggest that there is a complex relationship between cognitive and clinical factors in this high-risk population. PMID- 16714475 TI - Survival in hostile environments: strategies of renal medullary cells. AB - Cells in the renal medulla exist in a hostile milieu characterized by wide variations in extracellular solute concentrations, low oxygen tensions, and abundant reactive oxygen species. This article reviews the strategies adopted by these cells to allow them to survive and fulfill their functions under these extreme conditions. PMID- 16714476 TI - Wnt/Frizzled signaling in the vasculature: new angiogenic factors in sight. AB - Wnt growth factors function via Frizzled receptors to affect cellular proliferation, differentiation, apoptosis, and migration. Wnt/Frizzled signaling is now linked to human hereditary disorders with retinal vascular defects, implicating Wnts as angiogenic factors. Here, we discuss Wnts and a novel Frizzled ligand, Norrin, in physiological and pathological angiogenesis. PMID- 16714477 TI - Insulin vesicle release: walk, kiss, pause ... then run. AB - The mechanisms by which insulin-containing dense core secretory vesicles approach and finally fuse with the plasma membrane are of considerable current interest: defects in these processes may be one of the contributing factors to Type 2 diabetes. In this review, we discuss the molecular mechanisms involved in vesicle trafficking within the pancreatic beta-cell and the mechanisms whereby these may be regulated. We then go on to describe recent evidence that suggests that vesicle fusion at the plasma membrane is a partly reversible process ("kiss and run" or "cavity recapture"). We propose that vesicles may participate in a exo endocytotic cycle in which a proportion of those that have already undergone an interaction with the plasma membrane may exchange exocytotic machinery with maturing vesicles. PMID- 16714478 TI - Dynamic clamp analyses of cardiac, endocrine, and neural function. AB - The dynamic clamp introduces artificial conductances into cells to simulate electrical coupling, votage-dependent, leak, and synaptic conductances. This review describes how the dynamic clamp has been used to address various questions in the cardiac, endocrine, and nervous systems. PMID- 16714479 TI - Astrocytes coordinate synaptic networks: balanced excitation and inhibition. AB - Although neurons are essential for brain function, an emerging alternative view holds that astrocytes, the dominant glial cell type, coordinate synaptic networks. Through the release of glutamate, astrocytes locally excite neurons, and via adenosine, which accumulates due to the hydrolysis of released ATP, astrocytes suppress distant synapses. PMID- 16714480 TI - Neural bases of postural control. AB - The body posture during standing and walking is maintained due to the activity of a closed-loop control system. In the review, we consider different aspects of postural control: its functional organization, the distribution of postural functions in different parts of the central nervous system, and the activity of neuronal networks controlling posture. PMID- 16714484 TI - Multidrug resistance-associated proteins: expression and function in the central nervous system. AB - Drug delivery to the brain is highly restricted, since compounds must cross a series of structural and metabolic barriers to reach their final destination, often a cellular compartment such as neurons, microglia, or astrocytes. The primary barriers to the central nervous system are the blood-brain and blood cerebrospinal fluid barriers. Through structural modifications, including the presence of tight junctions that greatly limit paracellular transport, the cells that make up these barriers restrict diffusion of many pharmaceutically active compounds. In addition, the cells that comprise the blood-brain and blood cerebrospinal fluid barriers express multiple ATP-dependent, membrane-bound, efflux transporters, such as members of the multidrug resistance-associated protein (MRP) family, which contribute to lowered drug accumulation. A relatively new concept in brain drug distribution just beginning to be explored is the possibility that cellular components of the brain parenchyma could act as a "second" barrier to brain permeation of pharmacological agents via expression of many of the same transporters. Indeed, efflux transporters expressed in brain parenchyma may facilitate the overall export of xenobiotics from the central nervous system, essentially handing them off to the barrier tissues. We propose that these primary and secondary barriers work in tandem to limit overall accumulation and distribution of xenobiotics in the central nervous system. The present review summarizes recent knowledge in this area and emphasizes the clinical significance of MRP transporter expression in a variety of neurological disorders. PMID- 16714485 TI - Functional pharmacology in human brain. AB - Most neurological and psychiatric disorders involve selective or preferential impairments of neurotransmitter systems. Therefore, studies of functional transmitter pathophysiology in human brain are of unique importance in view of the development of effective, mechanism-based, therapeutic modalities. It is well known that central nervous system functional proteins, including receptors, transporters, ion channels, and enzymes, can exhibit high heterogeneity in terms of structure, function, and pharmacological profile. If the existence of types and subtypes of functional proteins amplifies the possibility of developing selective drugs, such heterogeneity certainly increases the likelihood of interspecies differences. It is therefore essential, before choosing animal models to be used in preclinical pharmacology experimentation, to establish whether functionally corresponding proteins in men and animals also display identical pharmacological profiles. Because of evidence that scaffolding proteins, trafficking between plasma membrane and intracellular pools, phosphorylation and allosteric modulators can affect the function of receptors and transporters, experiments with human clones expressed in host cells where the environment of native receptors is rarely reproduced should be interpreted with caution. Thus, the use of neurosurgically removed fresh human brain tissue samples in which receptors, transporters, ion channels, and enzymes essentially retain their natural environment represents a unique experimental approach to enlarge our understanding of human brain processes and to help in the choice of appropriate animal models. Using this experimental approach, many human brain functional proteins, in particular transmitter receptors, have been characterized in terms of localization, function, and pharmacological properties. PMID- 16714486 TI - Biological and biomaterial approaches for improved islet transplantation. AB - Islet transplantation may be used to treat type I diabetes. Despite tremendous progress in islet isolation, culture, and preservation, the clinical use of this modality of treatment is limited due to post-transplantation challenges to the islets such as the failure to revascularize and immune destruction of the islet graft. In addition, the need for lifelong strong immunosuppressing agents restricts the use of this option to a limited subset of patients, which is further restricted by the unmet need for large numbers of islets. Inadequate islet supply issues are being addressed by regeneration therapy and xenotransplantation. Various strategies are being tried to prevent beta-cell death, including immunoisolation using semipermeable biocompatible polymeric capsules and induction of immune tolerance. Genetic modification of islets promises to complement all these strategies toward the success of islet transplantation. Furthermore, synergistic application of more than one strategy is required for improving the success of islet transplantation. This review will critically address various insights developed in each individual strategy and for multipronged approaches, which will be helpful in achieving better outcomes. PMID- 16714487 TI - The ATP-binding cassette transporters and their implication in drug disposition: a special look at the heart. AB - The passage of drugs across cell membranes dictates their absorption, distribution, metabolism, and excretion. This process is determined by several factors including the molecular weight of the compounds, their shape, degree of ionization, and binding to proteins. Accumulation of xenobiotics into tissues does not depend only on their ability to enter cells, but also on their ability to leave them. For instance, the role of efflux transporters such as ATP-binding cassette (ABC) proteins in the disposition of drugs is now well recognized. Actually, ABC transporters act in synergy with drug-metabolizing enzymes to protect the organism from toxic compounds. The most studied transporter from the ABC transporter superfamily, P-glycoprotein, was found to be overexpressed in tumor cells and associated with an acquired resistance to several anticancer drugs. P-glycoprotein, thought at first to be confined to tumor cells, was subsequently recognized to be expressed in normal tissues such as the liver, kidney, intestine, and heart. Even though information remains rather limited on the functional role of ABC transporters in the myocardium, it is hypothesized that they may modulate efficacy and toxicity of cardioactive agents. This review addresses recent progress on knowledge about the ABC transporters in drug disposition and more precisely their role in drug distribution to the heart. PMID- 16714488 TI - Therapeutic targets: progress of their exploration and investigation of their characteristics. AB - Modern drug discovery is primarily based on the search and subsequent testing of drug candidates acting on a preselected therapeutic target. Progress in genomics, protein structure, proteomics, and disease mechanisms has led to a growing interest in and effort for finding new targets and more effective exploration of existing targets. The number of reported targets of marketed and investigational drugs has significantly increased in the past 8 years. There are 1535 targets collected in the therapeutic target database compared with approximately 500 targets reported in a 1996 review. Knowledge of these targets is helpful for molecular dissection of the mechanism of action of drugs and for predicting features that guide new drug design and the search for new targets. This article summarizes the progress of target exploration and investigates the characteristics of the currently explored targets to analyze their sequence, structure, family representation, pathway association, tissue distribution, and genome location features for finding clues useful for searching for new targets. Possible "rules" to guide the search for druggable proteins and the feasibility of using a statistical learning method for predicting druggable proteins directly from their sequences are discussed. PMID- 16714489 TI - "Punk" rock can be bad for you: a case of surgical emphysema from a "punk" rocker's leather jacket. AB - Stab wounds to the thorax are seen in the emergency department (ED) and can be caused by a variety of mechanisms. This case highlights an unusual cause of injury: a leather jacket with spikes on the back of it. This type of jacket is often worn by "punks" as a fashion statement. We report that falling onto such a jacket may result in accidental thoracic injury leading to subcutaneous emphysema. A thorough clinical assessment is mandatory to exclude underlying lung injury or pneumothorax. In patients with subcutaneous emphysema and an otherwise normal chest radiograph, an in hospital observation period of 24 hours to check for any delayed complications is adequate if the patient remains clinically stable. PMID- 16714490 TI - An unusual presentation of sphenoid sinusitis with septicaemia in a healthy young adult. AB - Streptococcus pneumonia and Haemophilus influenzae account for more than 50% of bacterial acute sinusitis. Isolated sphenoid sinusitis is a rare disease with potentially devastating complications such as cranial nerve involvement, brain abscess, and meningitis. It occurs at an incidence of about 2.7% of all sinus infections. There have been no previous reported cases of unilateral sphenoid sinusitis presenting as septicaemia in an otherwise healthy young immunocompetent adult. PMID- 16714491 TI - Anterior dislocation of the ulnar-humeral joint in a so-called 'pulled elbow'. AB - We describe an unusual case of a missed anterior dislocation of the elbow joint in a 1 year old girl who presented with a pulled elbow. To our knowledge, this is the first report of anterior dislocation as a result of a pulled elbow in the literature. We would like to highlight the rarity of this presentation and the importance of chronological assessment and management in the accident and emergency department. PMID- 16714492 TI - Atypical presentation of aortic dissection secondary to a gunshot wound. AB - We present a case of aortic dissection secondary to a gunshot wound. The traumatic aortic dissection in this patient occurred secondary to the cavitary forces produced by the bullet passing through the patient's chest. This is a rare phenomenon that could easily be missed if attention is placed only on the more obvious injuries. PMID- 16714493 TI - Atypical manifestation of Vibrio vulnificus septicaemia. AB - Vibrio vulnificus is a Gram-negative marine bacterium that may cause local wound infection, gastroenteritis, or septicaemia. Fatal septicaemia usually presents with fever, shock, and large haemorrhagic bullae on the legs. This report is about a man who had severe V. vulnificus septicaemia but presented with atypical features of leg pain and diffuse purpuric skin lesions. V. vulnificus septicaemia should be suspected if the following are present: septic shock, leg pains associated with diffuse purpuric skin lesions, recent consumption of raw seafood, and a past medical history of liver cirrhosis. PMID- 16714494 TI - An unusual rugby injury. AB - We describe an unusual sports injury in a young man, a combination of obturator hip dislocation with an ipsilateral anterior cruciate ligament injury. Traumatic non-prosthetic hip dislocations, particularly obturator hip dislocations, are extremely rare sports injuries and have not previously been reported in conjunction with a knee ligament injury. The severe pain and obvious deformity from the hip injury can distract from other injuries, particularly to the ipsilateral knee. This case reinforces the need for a thorough assessment of the knee joint, before hip reduction if possible and certainly after reduction of the dislocation. PMID- 16714495 TI - Rapid assessment teams and early discharge of the elderly from ED; vulnerable in the current financial climate? PMID- 16714496 TI - Alcoholic ketoacidosis. AB - Alcoholic ketoacidosis (AKA) is a common reason for investigation and admission of alcohol dependent patients in UK emergency departments. Although well described in international emergency medicine literature, UK emergency physicians rarely make the diagnosis of AKA. There is increasing evidence that rather than being benign and self limiting, AKA may be a significant cause of mortality in patients with alcohol dependence. This literature review discusses the history, characterisation, pathophysiology, diagnosis, and management of AKA. PMID- 16714497 TI - Clinical management of casualties exposed to lung damaging agents: a critical review. AB - There is no specific antidote for the treatment of casualties exposed to chlorine, phosgene, or mustards; therefore, management is largely supportive. Corticosteroid treatment has been given to casualties accidentally exposed to chlorine. Clinical data on efficacy are inconclusive as the numbers given steroids have been small and the indications for administration unclear. There have been no clinical controlled studies. There is a stronger evidence base from animal studies, particularly from porcine and rodent models. Lung injury induced by phosgene and mustard appears to be mediated by glutathione depletion, lipid peroxidation, free radical generation, and subsequent cellular toxicity. There is limited evidence to suggest that repletion of glutathione reduces and/or prevents lung damage by these agents. This may provide an opportunity for therapeutic intervention. PMID- 16714498 TI - Confidential registration in health services: randomised controlled trial. AB - BACKGROUND: Human rights legislation safeguards the privacy and dignity of patients. OBJECTIVE: To assess the effectiveness in terms of patient assessed privacy of confidential registration. DESIGN: Randomised controlled trial. SETTING: Emergency Department, University Hospital of Wales. PARTICIPANTS: A total of 302 patients aged over 15 years. MAIN OUTCOME MEASURES: Binary choices and ordinal visual analogue scores from a validated questionnaire on self reported measures: patient ability and preference to speak to receptionists and disclose confidential information without being overhead and concern about disclosure of items of confidential personal information. RESULTS: Patients who registered in a screened area felt significantly more able to tell receptionists things they did not want others to hear. Control patients were significantly more concerned than intervention patients that others heard their name, address, date of birth, reason for emergency department attendance, and telephone number, but not their marital status. Overall, intervention patients were less concerned about disclosure of information and that they had been overheard. CONCLUSIONS: Patients value privacy when they register and are concerned if others can hear them tell receptionists who they are, how to contact them, and why they are there. Confidential registration should be instituted in health services. Confidential registration increased patient privacy and should be instituted in health services. PMID- 16714499 TI - Alcohol related conditions represent a major psychiatric problem in emergency departments. AB - BACKGROUND: Alcoholism represents a huge socioeconomic burden in most developed societies. METHODS: In order to determine the impact of conditions associated with alcohol abuse on emergency care, in 2002 all patients with alcohol related pathology presenting to the emergency department (ED) of a large urban university hospital in Germany were assessed. RESULTS: Of 2194 patients seen by psychiatrists in the ED, the 613 cases associated with alcohol abuse represented the largest diagnostic group (30.7%). Within this group, alcohol intoxication was the most frequent diagnosis (71.4%). Patients with an alcohol related condition needed treatment by different medical specialities and required diagnostic procedures significantly more often and consequently stayed significantly longer in the ED than patients with other psychiatric presentations. CONCLUSIONS: The generally underestimated problem of alcohol abuse in Germany demands an excessive amount of manpower and resources in EDs, where a high yet expensive standard of care is provided. Against this background, how far this burden can be reduced, both in EDs and in society in general, is discussed. PMID- 16714500 TI - Violence and aggression in the emergency department. AB - OBJECTIVE: To investigate the characteristics of incidents of aggression and violence directed towards staff in an urban UK emergency department. METHODS: A retrospective review of incident report forms submitted over a 1 year period that collected data pertaining to the characteristics of assailants, the outcome of incidents, and the presence of possible contributory factors. RESULTS: A total of 218 incident reports were reviewed. It was found that the majority of assailants were patients, most were male, and the median age was 32 years. Assailants were more likely to live in deprived areas than other patients and repeat offenders committed 45 of the incidents reported during the study period. The incident report indicated that staff thought the assailant was under the influence of alcohol on 114 occasions. Incidents in which the assailant was documented to have expressed suicidal ideation or had been referred to the psychiatric services were significantly more likely to describe physical violence, as were those incidents in which the assailant was female. CONCLUSION: Departments should seek to monitor individuals responsible for episodes of violence and aggression in order to detect repeat offenders. A prospective study comprising post-incident reviews may provide a valuable insight into the causes of violence and aggression. PMID- 16714501 TI - The evolution of the emergency care practitioner role in England: experiences and impact. AB - BACKGROUND: The emergency care practitioner (ECP) is a generic practitioner who combines extended nursing and paramedic skills. The "new" role emerged out of changing workforce initiatives intended to improve staff career opportunities in the National Health Service and ensure that patients' health needs are assessed appropriately. OBJECTIVE: To describe the development of ECP Schemes in 17 sites, identify criteria contributing to a successful operational framework, analyse routinely collected data and provide a preliminary estimate of costs. METHODS: There were three methods used: (a) a quantitative survey, comprising a questionnaire to project leaders in 17 sites, and analysis of data collected routinely; (b) qualitative interpretation based on telephone interviews in six sites; and (c) an economic costing study. RESULTS: Of 17 sites, 14 (82.5%) responded to the questionnaire. Most ECPs (77.4%) had trained as paramedics. Skills and competencies have been extended through educational programmes, training, and assessment. Routine data indicate that 54% of patient contacts with the ECP service did not require a referral to another health professional or use of emergency transport. In a subset of six sites, factors contributing to a successful operational framework were strategic visions crossing traditional organisational boundaries and appropriately skilled workforce integrating flexibly with existing services. Issues across all schemes were patient safety, appropriate clinical governance, and supervision and workforce issues. On the data available, the mean cost per ECP patient contact is 24.00 pounds sterling, which is less than an ED contact of 55.00 pounds sterling. CONCLUSION: Indications are that the ECP schemes are moving forward in line with original objectives and could be having a significant impact on the emergency services workload. PMID- 16714502 TI - Variability in pupil size estimation. AB - BACKGROUND: The clinical estimation of pupil size and reactivity is central to the neurological assessment of patients, particularly those with or at risk of neurological damage. Health care professionals who examine pupils have differing levels of skill and training, yet their recordings are passed along the patient care pathway and can influence care decisions. The aim of this study was to determine if any statistical differences existed in the estimation of pupil size by different groups of health care professionals. METHODS: A total of 102 health care professionals working in the critical care environment were asked to estimate and record the pupil size of a series of 12 artificial eyes with varying pupil diameter and iris colour. All estimations were performed indoors under ambient lighting conditions. RESULTS: Our results established a statistically significant difference between staff groups in the estimation of pupil size. CONCLUSION: The demonstrated variability in pupil size estimation may not be clinically significant. However, it remains desirable to have consistency of measurement throughout the patient care pathway. PMID- 16714503 TI - When is a door not a door? The difference between documented and actual arrival times in the emergency department. AB - OBJECTIVES: An observational study to determine the difference between documented ambulance arrival times and the actual arrival times of patients from the ambulance into the emergency department. METHODS: In a busy, purpose built, modern emergency department with easy access, we recorded the time that ambulance borne patients were wheeled over the threshold of the clinical area and compared this to the times recorded by the ambulance trusts as the official ambulance arrival times. RESULTS: 352 ambulance arrivals were observed. Data were incomplete in 34 instances (9.5%) and were not included in the analysis. For the remaining 318 arrivals, the median time difference was 2 min 1 s (range 5 s to 21 min 45 s). In a subgroup of chest pain patients (45 patients), the median time difference was 2 min 11 s (range 23 s to 5 min 38 s). The difference between the chest pain group and the remaining patients was not significant (p = 0.528). CONCLUSIONS: There is inevitably some delay between the arrival of an ambulance and the arrival of the patient into a clinical area. This study quantifies that difference. In an era of stringent time related standards, this paper highlights the need for accurate recording of times to enable us to carry out valid audit of these standards. This study supports the redefining of an arrival time as the time when the patient arrives in the clinical area. PMID- 16714504 TI - What percentages of patients are suitable for prehospital thrombolysis? AB - OBJECTIVE: To apply the Joint Royal College Ambulance Liaison Committee (JRCALC) checklist to patients who were deemed eligible for thrombolytic therapy on arrival in an Accident & Emergency Department (A&E) to determine the proportion suitable for prehospital thrombolysis. DESIGN: Retrospective descriptive analysis. METHODS: The clinical notes of all patients thrombolysed in an A&E department in a year were reviewed against the JRCALC guidelines for prehospital thrombolysis. RESULTS: 14.2% of patients eligible for thrombolysis in a district general hospital were deemed suitable for prehospital thrombolysis according to the JRCALC criteria. The most common exclusion criteria were hyper/hypotension (50%), onset of symptoms (pain) >6 h previously (41.7%), or age >75 years (37%). Two or more contraindications to prehospital thrombolysis were present in 63.9% of patients. CONCLUSION: The JRCALC guidelines are an effective tool for identifying patients with potential contraindications to thrombolysis. PMID- 16714505 TI - Evaluation of focussed assessment with sonography in trauma (FAST) by UK emergency physicians. AB - OBJECTIVES: To evaluate the introduction of a focussed assessment with sonography in trauma (FAST) scan into the early assessment of trauma patients in the UK. METHODS: The setting was an inner city teaching hospital emergency department (annual attendance 100,000). All patients aged 16 or over admitted to the resuscitation room after blunt trauma were included in a prospective observational study. Patients had a FAST scan performed at the end of the primary survey. Results were compared to results of other investigations, laparotomy, postmortem examination, or observation. RESULTS: 153 patients were entered into the study. The sensitivity of the FAST scan was 78% and specificity was 99%. CONCLUSION: FAST is a highly specific "rule in" technique and is useful in the initial assessment of trauma patients. Emergency physicians can perform FAST after a brief training period. PMID- 16714506 TI - Evaluation of a diabetes referral pathway for the management of hypoglycaemia following emergency contact with the ambulance service to a diabetes specialist nurse team. AB - OBJECTIVES: To evaluate the effectiveness of ambulance crew referral of patients treated for acute hypoglycaemia, to a diabetes specialist nurse (DSN) team. Patients were assessed by the ambulance service and did not attend an emergency department. METHODS: For a 3 month period patients were referred from two areas of West Yorkshire by the ambulance service to specialist diabetes teams. The DSNs contacted the patients within 7 days by telephone and arranged further review if necessary. Satisfaction questionnaires were sent to patients. RESULTS: Thirty eight patients were referred and all were reviewed by telephone; 30 (79%) also required a clinic review appointment. Warning signs of hypoglycaemia were identified by 21 (55%) patients, but 14 (37%) had none (three had incomplete data). Twelve patients had had a self-treated episode in the previous 6 months; nine patients had had three or more. Fifteen (39%) patients had called an emergency ambulance for similar reasons in the previous 6 months (27 "999" calls). Twenty patients (53%) had their drug treatment altered at a clinic appointment and 14 (37%) required ongoing review. Twenty six patients returned the satisfaction questionnaire; of these 88% agreed or strongly agreed that they had improved their understanding of hypoglycaemia and 73% felt more able to treat a hypoglycaemic episode in the future. Patient satisfaction with this care pathway was high. CONCLUSIONS: The ambulance service can coordinate successful referral of patients with episodes of acute hypoglycaemia to a DSN-led service. Patients often required treatment review. They valued this service and felt more confident managing their diabetes. PMID- 16714507 TI - Preventable deaths in patients admitted from emergency department. AB - BACKGROUND: There is limited data about how appropriate medical care is in the emergency department (ED). OBJECTIVES: To investigate the rate and types of preventable deaths among patients with early mortality after emergency admission from the ED. METHODS: We retrospectively reviewed charts of early mortality (defined as mortality which occurred within 24 hours after admission from the ED) over a 3 year period. Those patients with terminal cancer or out of hospital cardiac arrest (OHCA) at presentation were excluded. Two independent assessors reviewed each eligible chart and determined whether early mortality was preventable. Any disagreements were resolved through discussion between the investigators. A mortality event was considered preventable if actions or missed actions were identified that would have prevented the death. The types of preventability were categorised as misdiagnosis, delayed diagnosis, and inappropriate medical management. Interrater reliability in the initial determination was assessed using Cohen kappa statistic. RESULTS: Over a 3 year period, 210 early mortality cases were identified. Excluding patients with terminal cancer or OHCA, the rate of preventable deaths was 25.8% (32/124). The types of preventability were inappropriate medical management (17 patients), delayed diagnosis (eight), and misdiagnosis (seven). There was good agreement between assessors with a Cohen kappa statistic of 0.81. CONCLUSIONS: Preventable deaths in emergency admitted patients with early mortality are not uncommon. Analysis and identification of preventability early mortality by using a chart based method may be used as a quality assurance index in emergency medical care. PMID- 16714508 TI - Things that go beep: experience with an ED guideline for use of a handheld metal detector in the management of ingested non-hazardous metallic foreign bodies. AB - OBJECTIVES: To review compliance with our emergency department (ED) guideline on the imaging of ingested non-hazardous metallic foreign bodies in children, investigate adverse outcomes, and make suggestions for improving the guideline. METHODS: Retrospective analysis of patients presenting in a 3 year period to a paediatric ED with a history of possible metallic foreign body (MFB) ingestion, who were managed according to an ED guideline. RESULTS: We identified 430 episodes of possible MFB ingestion, of which 422 were eligible for inclusion in the study. Compliance with the guideline was 77.8% with no significant adverse events. The exclusion of symptoms as a criterion for x ray results in a reduction in the x ray rate of 56% in the symptomatic group with no increase in adverse events. CONCLUSION: A handheld metal detector (HMD) can be safely and reliably used in lieu of plain radiography to investigate children with a history of MFB ingestion, irrespective of symptoms and without incurring any significant adverse events. PMID- 16714509 TI - The new consultant survey 2005. AB - BACKGROUND: Consultants in emergency medicine have to deal with a wide range of problems, many of which they will not have encountered during their training. One way to assess the adequacy of specialist training is to ask recently appointed consultants whether or not they feel adequately prepared for their role. METHODS: A questionnaire was sent out to 60 newly appointed consultants in emergency medicine in January and February 2005 and the results analysed. RESULTS: Many respondents feel that there should be greater emphasis on acquiring clinical skills, partly by greater consultant supervision and partly by providing more experience of anaesthetics and intensive care. New consultants also feel inadequately prepared for their management responsibilities, and this is a source of great stress. CONCLUSIONS: Specialist training in emergency medicine needs to pay more attention to the acquisition of clinical skills and to preparation for management responsibility. PMID- 16714510 TI - Retroperitoneal haemorrhage caused by a renal angiomyolipoma. AB - Renal angiomyolipoma (AML) is a benign renal tumour and is nowadays considered a relatively common lesion. When an AML increases in size or becomes symptomatic, embolisation via the renal artery should then be considered, because rupture is an important complication and interventional therapies are required to stop bleeding. We present a 21 year old woman who was seen at the emergency department following a low velocity trauma. After a period of 9 weeks, clinical examination and radiological examination revealed a haemorrhage from a renal AML, which was treated by selective embolisation. A discussion of the relevant literature is also presented. PMID- 16714511 TI - Spontaneous pneumopericardium, pneumomediastinum and subcutaneous emphysema: unusual complications of asthma in a 2-year-old boy. AB - A 2-year-old boy presented to the emergency department with a history of sudden onset of cough, dyspnoea and a slight expiratory wheeze on the right lung base. He also had subcutaneous emphysema on the left side of the chest anteriorly. Chest x ray confirmed subcutaneous emphysema and also revealed pneumomediastinum and pneumopericardium. He had had no previous episode and was not known to have asthma. He was apyrexial but had a raised white cell count. The eosinophil count was within normal limits. He was successfully treated with nebulised salbutamol, steroids, antibiotics and high flow oxygen. He made a good recovery and was discharged after 7 days. This case highlights the need for a high index of suspicion of asthma in very young children presenting for the first time with such complications. PMID- 16714512 TI - Mental health service requirements in a Japanese medical centre emergency department. AB - BACKGROUND: A significant number of emergency department (ED) patients in Japan may be affected by mental health problems leading to attempted suicide. This retrospective descriptive study aimed to explore mental health service needs in a Japanese medical centre ED. METHODS: Data on all inpatients were collected within 24 h of admission to the ED of a medical centre in Tokyo from 1st February 2004 to the 31st of January 2005. RESULTS: A total of 14.4% (95% CI 13.0 to 16.0) of ED visits required psychiatric services and 23.4% (95% CI 21.7 to 25.3) of inpatients had experienced psychologically traumatic events. CONCLUSIONS: Approximately 38% (95% CI 35.8 to 39.9) of patients presenting to an ED could be affected by mental health problems. PMID- 16714513 TI - Best evidence topic reports. Beta-agonists with or without anti-cholinergics in the treatment of acute childhood asthma? PMID- 16714515 TI - Best evidence topic reports. Delivery of bronchodilators in acute asthma in children. PMID- 16714516 TI - Best evidence topic reports. Lorazepam or diazepam in paediatric status epilepticus. PMID- 16714517 TI - Best evidence topic reports. Tibial fractures in very young children and child abuse. PMID- 16714518 TI - Comparison of paediatric major incident primary triage tools. AB - OBJECTIVES: To determine the sensitivity and specificity of paediatric major incident triage scores. The Paediatric Triage Tape (PTT), Careflight, Simple Triage and Rapid Treatment (START), and JumpSTART systems were tested. METHODS: In total, 3461 children presenting to a South African emergency department with trauma were scored using the four different methods. The sensitivity and specificity of the four scores was calculated against the Injury Severity Score (ISS), New ISS (NISS), and a modification of the Garner criteria (a measure of need for urgent clinical intervention). We also performed a Bayesian analysis of the scores against three different types of major incident. RESULTS: None of the tools showed high sensitivity and specificity. Overall, the Careflight score had the best performance in terms of sensitivity and specificity. The performance of the PTT was very similar. In contrast, the JumpSTART and START scores had very low sensitivities, which meant that they failed to identify patients with serious injury, and would have missed the majority of seriously injured casualties in the models of major incidents. CONCLUSION: The Careflight or PTT methods of triage should be used in paediatric major incidents in preference to the jumpSTART or START methods. PMID- 16714519 TI - The costs of falls in the community to the North East Ambulance Service. AB - BACKGROUND: This study set out to quantify the immediate costs to the North East Ambulance Service (NEAS) of attending to fallers. METHODS: Data from the Newcastle, UK area were collated by NEAS to identify those aged over 65 who had fallen and required an assistance only call or were subsequently transported to an Accident and Emergency (A&E) department. The 2001 census data for the total population served by NEAS in Newcastle were obtained. RESULTS: The total population of Newcastle over the age of 65 was 41,338. Over 7 months NEAS attended to 1504 falls in Newcastle (at 115 pounds sterling per call out, this equates to 172,960 pounds sterling). The faller was transported to A&E on 1339 occasions, while 165 falls required assistance only (11% of total) (36 falls requiring NEAS assistance per 1000 Newcastle population aged over 65 in 7 months). The total time on site for ambulance crews attending to fallers was 377.1 h (15.7 days in 7 months or 2.25 days per month). As the cost of emergency ambulance time is 123 pounds sterling/h, the total cost was 46,383.30 pounds sterling. Therefore, in Newcastle, attending to fallers in the community costs NEAS 376,018 pounds sterling per year (145.83 pounds sterling per fall or 9.10 pounds sterlingper person over the age of 65 per year). CONCLUSION: NEAS attend to a significant number of older people who fall in the community. In Newcastle alone the cost of this service equates to over 2 days of emergency ambulance crew time per month. Studies are needed to determine whether responding to falls in the community differently would be cost effective. PMID- 16714521 TI - Case of the month: Buffalo chest: a case of bilateral pneumothoraces due to pleuropleural communication. AB - Simultaneous spontaneous bilateral pneumothoraces, the presentation of separate right and left pneumothoraces together, is a rare event. The pleural cavities in humans are separated completely and the only previous reports of pleuropleural communication have been associated with major invasive thoracic procedures, specifically mediastinal surgery. The term "buffalo chest" has been coined for the condition on the basis that the buffalo or bison has a single pleural cavity, one of the few mammals to do so. We present the case of a woman with a past history of a single right sided spontaneous pneumothorax but no major thoracic surgery, who presented to the emergency department with a second spontaneous right pneumothorax that was under tension. After thoracostomy, she was found to have bilateral pneumothoraces which resolved with a unilateral chest drain demonstrating pleuropleural communication. We believe this to be the first reported case of such a presentation in the literature. The case demonstrates an unusual emergency presentation of a rare anatomical anomaly. PMID- 16714523 TI - Unusual way of purging. PMID- 16714524 TI - Are chest pain observation units essential for rapid and effective emergency care in the UK? PMID- 16714525 TI - Placement confirmation of Sengstaken-Blakemore tube by ultrasound. PMID- 16714526 TI - "Street" supplies of the anticoagulant drug warfarin: a worrying new trend. PMID- 16714527 TI - Intranasal alfentanil for acute pain in children. PMID- 16714528 TI - Injury to supraorbital area--should this be referred to as a head injury or facial injury? PMID- 16714529 TI - Consent in emergency research. PMID- 16714530 TI - Prevalence of psychological distress assessed in emergency departments. PMID- 16714531 TI - Prehospital Intubation--delving deeper into the evidence. PMID- 16714532 TI - Association does not prove causality. PMID- 16714534 TI - Please don't shoot the messengers! PMID- 16714533 TI - Best BETS: a call for scrutiny. PMID- 16714535 TI - Of mice and men, revisited: new insights into an ancient molecule from studies of complement activation by Cryptococcus neoformans. PMID- 16714537 TI - Differential localization of complement component 3 within the capsular matrix of Cryptococcus neoformans. AB - The polysaccharide capsule of Cryptococcus neoformans is a powerful activator of the complement system. The goal of the present study was to assess serum and cellular variables that influence the sites for C3 binding within the capsular matrix. Confocal microscopy using fluorophore-labeled polyclonal anti-C3 and anticapsular monoclonal antibodies and rosetting of fluorescent microspheres coated with anti-C3 were used to identify sites of C3 binding relative to the capsular edge. The results showed that the source of serum was a major variable influencing localization of C3. C3 bound at or very near the capsular edge in the case of human serum. C3 deposition was further from the capsule edge with guinea pig and rat sera; in the case of mouse serum, there was no binding of C3 in the outer region of the capsule. Addition of human C3 to mouse serum led to deposition of the C3 at the capsular edge, indicating that distinct properties of mouse and human C3 account for the differential localization of C3. Finally, the density of the capsular matrix was an important variable in determining sites for C3 deposition. Yeast cells with a high concentration of polysaccharide near the capsule edge supported deposition of mouse C3 at or near the capsular edge, whereas cells with a low matrix density showed deposition well beneath the edge. Taken together, these results indicate that the spatial deposition of C3 within the capsular matrix is a complex process that is influenced by the serum source and the density of the capsular matrix. PMID- 16714536 TI - Interaction of neonatal phagocytes with group B streptococcus: recognition and response. PMID- 16714538 TI - Significance of heat-stable and heat-labile enterotoxins in porcine colibacillosis in an additive model for pathogenicity studies. AB - Although heat-stable (ST) and heat-labile (LT) enterotoxins produced by enterotoxigenic Escherichia coli (ETEC) have been documented as important factors associated with diarrheal diseases, investigations assessing the contributions of individual enterotoxins to the pathogenesis of E. coli infection have been limited. To address the individual roles of enterotoxins in the diarrheal disease caused by K88-positive ETEC in young pigs, enterotoxin-positive and -negative isogenic E. coli strains were constructed by using pBR322 to clone and express LT and STb. Four strains, K88+ astA, K88+ astA/pBR322, K88+ astA STb+, and K88+ astA LT+, were constructed and subsequently included in gnotobiotic piglet challenge studies, and their pathogenesis was assessed. The results indicated that all K88+ isogenic strains were able to colonize the small intestines of piglets exhibiting the K88 receptor. However, only LT- and STb-positive strains caused appreciable diarrhea. Piglets inoculated with the K88+ astA LT+ strain became dehydrated within 18 h, while those inoculated with the K88+ astA STb+ strain did not, although diarrhea developed in several piglets. The changes in the blood packed cell volume and plasma total protein of gnotobiotic piglets inoculated with the LT-positive strains were significantly greater than those of pigs inoculated with the K88 astA/pBR322 strain (P = 0.012, P = 0.002). Immunochemistry image analysis also suggested that LT enhanced bacterial colonization in a gnotobiotic piglet model. This investigation suggested that LT is a major contributor to the virulence of K88+ ETEC and that isogenic constructs are a useful tool for studying the pathogenesis of ETEC infection. PMID- 16714539 TI - Role for erbin in bacterial activation of Nod2. AB - Intracellular peptidoglycan (PG) recognition in human cells is mediated by the NACHT-LRR proteins Nod1 and Nod2. Elicitation of these proteins by PG motifs released from invasive bacteria triggers signaling events, resulting in the activation of the NF-kappaB pathway. In order to decipher the molecular components involved in Nod2 signal transduction, we set out to identify new interaction partners of Nod2 by using a yeast two-hybrid screen. Besides the known interaction partner RIP2, the screen identified the leucine-rich repeat (LRR)- and PDZ domain-containing family member Erbin as a binding partner of Nod2. Erbin showed a specific interaction with Nod2 in coimmunoprecipitation experiments with human HEK 293T cells. Immunofluorescence microscopy with a newly generated anti-Nod2 monoclonal antibody showed that Erbin and Nod2 partially colocalize in human cells. Subsequent analysis of the Erbin/Nod2 interaction revealed that the LRR of Erbin and the caspase activating and recruiting domains of Nod2 were necessary for this interaction. No significant interaction was observed with a Walker B box mutant of Nod2 or a Crohn's disease-associated frameshift mutant of Nod2, indicating that complex formation is dependent on the activity of the molecule. In addition, a change in the dynamics of the Erbin/Nod2 complex was observed during Shigella flexneri infection. Furthermore, ectopic expression of increasing amounts of Erbin or short hairpin RNA-mediated knockdown of Erbin showed a negative influence of Erbin on Nod2/muramyl-dipeptide-mediated NF-kappaB activation. These results implicate Erbin as a potential negative regulator of Nod2 and show that bacterial infection has an impact on Nod2/Erbin complex formation within cells. PMID- 16714540 TI - Mycobacterium marinum Erp is a virulence determinant required for cell wall integrity and intracellular survival. AB - The Mycobacterium tuberculosis exported repetitive protein (Erp) is a virulence determinant required for growth in cultured macrophages and in vivo. To better understand the role of Erp in Mycobacterium pathogenesis, we generated a mutation in the erp homologue of Mycobacterium marinum, a close genetic relative of M. tuberculosis. erp-deficient M. marinum was growth attenuated in cultured macrophage monolayers and during chronic granulomatous infection of leopard frogs, suggesting that Erp function is similarly required for the virulence of both M. tuberculosis and M. marinum. To pinpoint the step in infection at which Erp is required, we utilized a zebrafish embryo infection model that allows M. marinum infections to be visualized in real-time, comparing the erp-deficient strain to a DeltaRD1 mutant whose stage of attenuation was previously characterized in zebrafish embryos. A detailed microscopic examination of infected embryos revealed that bacteria lacking Erp were compromised very early in infection, failing to grow and/or survive upon phagocytosis by host macrophages. In contrast, DeltaRD1 mutant bacteria grow normally in macrophages but fail to induce host macrophage aggregation and subsequent cell-to-cell spread. Consistent with these in vivo findings, erp-deficient but not RD1 deficient bacteria exhibited permeability defects in vitro, which may be responsible for their specific failure to survive in host macrophages. PMID- 16714542 TI - Resistance of Streptococcus gordonii to polymorphonuclear leukocyte killing is a potential virulence determinant of infective endocarditis. AB - Significant differences in virulence among seven representative Streptococcus gordonii strains were observed by using the rat model of infective endocarditis. Five strains, including S. gordonii DL1, caused severe disease, while the other two strains, including S. gordonii SK12, caused minimal or no disease. The differences in virulence were evident from the visible presence of streptococci in the vegetations present on the aortic valves of catheterized rats that were challenged with individual strains and also from the much greater recovery of rifampin-resistant S. gordonii DLl than of streptomycin-resistant S. gordonii SK12 from the hearts of animals coinfected with both organisms. Each S. gordonii strain aggregated with human platelets and bound to polymorphonuclear leukocytes (PMNs), as shown by the stimulation of PMN superoxide anion production. These interactions were reduced or abolished by pretreatment of the platelets or PMNs with sialidase, indicating that there was bacterial recognition of host sialic acid-containing receptors. Adhesin-mediated binding of each S. gordonii strain to PMNs also triggered phagocytosis. However, the subsequent PMN-dependent killing differed significantly for the seven strains. The five virulent strains included three strains that were not killed and two strains whose numbers were reduced by approximately 50%. In contrast, the level of killing of each avirulent strain under the same conditions was significantly greater and approached 90% of the bacteria added. Parallel studies performed with rat PMNs revealed comparable differences in the resistance or susceptibility of representative virulent and avirulent strains. Thus, the ability of S. gordonii to survive in PMNs following adhesin-mediated phagocytosis may be an important virulence determinant of infective endocarditis. PMID- 16714541 TI - Rhamnolipids are virulence factors that promote early infiltration of primary human airway epithelia by Pseudomonas aeruginosa. AB - The opportunistic bacterium Pseudomonas aeruginosa causes chronic respiratory infections in cystic fibrosis and immunocompromised individuals. Bacterial adherence to the basolateral domain of the host cells and internalization are thought to participate in P. aeruginosa pathogenicity. However, the mechanism by which the pathogen initially modulates the paracellular permeability of polarized respiratory epithelia remains to be understood. To investigate this mechanism, we have searched for virulence factors secreted by P. aeruginosa that affect the structure of human airway epithelium in the early stages of infection. We have found that only bacterial strains secreting rhamnolipids were efficient in modulating the barrier function of an in vitro-reconstituted human respiratory epithelium, irrespective of their release of elastase and lipopolysaccharide. In contrast to previous reports, we document that P. aeruginosa was not internalized by epithelial cells. We further report that purified rhamnolipids, applied on the surfaces of the epithelia, were sufficient to functionally disrupt the epithelia and to promote the paracellular invasion of rhamnolipid-deficient P. aeruginosa. The mechanism involves the incorporation of rhamnolipids within the host cell membrane, leading to tight-junction alterations. The study provides direct evidence for a hitherto unknown mechanism whereby the junction-dependent barrier of the respiratory epithelium is selectively altered by rhamnolipids. PMID- 16714543 TI - Salmonella enterica serovar Typhimurium requires the Lpf, Pef, and Tafi fimbriae for biofilm formation on HEp-2 tissue culture cells and chicken intestinal epithelium. AB - Recent work has demonstrated that Salmonella enterica serovar Typhimurium forms biofilms on HEp-2 tissue culture cells in a type 1 fimbria-dependent manner. To investigate how biofilm growth of HEp-2 tissue culture cells affects gene expression in Salmonella, we compared global gene expression during planktonic growth and biofilm growth. Microarray results indicated that the transcription of approximately 100 genes was substantially altered by growth in a biofilm. These genes encode proteins with a wide range of functions, including antibiotic resistance, central metabolism, conjugation, intracellular survival, membrane transport, regulation, and fimbrial biosynthesis. The identification of five fimbrial gene clusters was of particular interest, as we have demonstrated that type 1 fimbriae are required for biofilm formation on HEp-2 cells and murine intestinal epithelium. Mutations in each of these fimbriae were constructed in S. enterica serovar Typhimurium strain BJ2710, and the mutants were found to have various biofilm phenotypes on plastic, HEp-2 cells, and chicken intestinal tissue. The pef and csg mutants were defective for biofilm formation on each of the three surfaces tested, while the lpf mutant exhibited a complete loss of the ability to form a biofilm on chicken intestinal tissue but only an intermediate loss of the ability to form a biofilm on tissue culture cells and plastic surfaces. The bcf mutant displayed increased biofilm formation on both HEp-2 cells and chicken intestinal epithelium, while the sth mutant had no detectable biofilm defects. In all instances, the mutants could be restored to a wild-type phenotype by a plasmid carrying the functional genes. This is the first work to identify the genomic responses of Salmonella to biofilm formation on host cells, and this work highlights the importance of fimbriae in adhering to and adapting to a eukaryotic cell surface. An understanding of these interactions is likely to provide new insights for intervention strategies in Salmonella colonization and infection. PMID- 16714544 TI - Macrophage migration inhibitory factor contributes to host defense against acute Trypanosoma cruzi infection. AB - Macrophage migration inhibitory factor (MIF) is a proinflammatory cytokine that is involved in the host defense against several pathogens. Here we used MIF-/- mice to determine the role of endogenous MIF in the regulation of the host immune response against Trypanosoma cruzi infection. MIF-/- mice displayed high levels of blood and tissue parasitemia, developed severe heart and skeletal muscle immunopathology, and succumbed to T. cruzi infection faster than MIF+/+ mice. The enhanced susceptibility of MIF-/- mice to T. cruzi was associated with reduced levels of proinflammatory cytokines, such as tumor necrosis factor alpha, interleukin-12 (IL-12), IL-18, gamma interferon (IFN-gamma), and IL-1beta, in their sera and reduced production of IL-12, IFN-gamma, and IL-4 by spleen cells during the early phase of infection. At all time points, antigen-stimulated splenocytes from MIF+/+ and MIF-/- mice produced comparable levels of IL-10. MIF /- mice also produced significantly less Th1-associated antigen-specific immunoglobulin G2a (IgG2a) throughout the infection, but both groups produced comparable levels of Th2-associated IgG1. Lastly, inflamed hearts from T. cruzi infected MIF-/- mice expressed increased transcripts for IFN-gamma, but fewer for IL-12 p35, IL-12 p40, IL-23, and inducible nitric oxide synthase, compared to MIF+/+ mice. Taken together, our findings show that MIF plays a role in controlling acute T. cruzi infection. PMID- 16714545 TI - Differential requirements for soluble and transmembrane tumor necrosis factor in the immunological control of primary and secondary Listeria monocytogenes infection. AB - The relative contributions of transmembrane tumor necrosis factor (memTNF) and soluble tumor necrosis factor (solTNF) in innate and adaptive immunity are poorly defined. We examined the capacities of wild-type (WT) mice, TNF-/- mice, and memTNF mice, which express only transmembrane TNF, to control primary and secondary Listeria monocytogenes infections. Soluble TNF was not required for induction or maintenance of protective immunity against a low-dose (200-CFU) Listeria infection. In contrast to TNF-/- mice, both WT and memTNF mice cleared the bacilli within 10 days and were fully protected against rechallenge with a lethal infective dose. Furthermore, T cells transferred from immune mice, but not from naive, WT, and memTNF mice, protected TNF-/- recipients against an otherwise lethal infection. By contrast, infection with a higher dose of Listeria (2,000 CFU) clearly demonstrated that solTNF is required to coordinate an optimal protective inflammatory response. memTNF mice were more susceptible to a high dose infection, and they exhibited delayed bacterial clearance, increased inflammation, and necrosis in the liver that resulted in 55% mortality. The dysregulated inflammation was accompanied by prolonged elevated expression of mRNAs for several chemokines as well as the macrophage effector molecules inducible nitric oxide synthase and LRG-47 in the livers of memTNF mice but not in the livers of WT mice. These data demonstrated that memTNF is sufficient for establishing protective immunity against a primary low-dose Listeria infection but that solTNF is required for optimal control of cellular inflammation and resistance to a primary high-dose infection. By contrast, memTNF alone is sufficient for resolution of a secondary, high-dose infection and for the transfer of protective immunity with memory T cells. PMID- 16714546 TI - Involvement of gonadal steroids and gamma interferon in sex differences in response to blood-stage malaria infection. AB - To examine the hormonal and immunological mechanisms that mediate sex differences in susceptibility to malaria infection, intact and gonadectomized (gdx) C57BL/6 mice were inoculated with Plasmodium chabaudi AS-infected erythrocytes, and the responses to infection were monitored. In addition to reduced mortality, intact females recovered from infection-induced weigh loss and anemia faster than intact males. Expression microarrays and real-time reverse transcription-PCR revealed that gonadally intact females exhibited higher expression of interleukin-10 (IL 10), IL-15Ralpha, IL-12Rbeta, Gadd45gamma, gamma interferon (IFN-gamma), CCL3, CXCL10, CCR5, and several IFN-inducible genes in white blood cells and produced more IFN-gamma than did intact males and gdx females, with these differences being most pronounced during peak parasitemia. Intact females also had higher anti-P. chabaudi immunoglobulin G (IgG) and IgG1 responses than either intact males or gdx females. To further examine the effector mechanisms mediating sex differences in response to P. chabaudi infection, responses to infection were compared among male and female wild-type (WT), T-cell-deficient (TCRbetadelta-/ ), B-cell-deficient (microMT), combined T- and B-cell-deficient (RAG1), and IFN gamma knockout (IFN-gamma-/-) mice. Males were 3.5 times more likely to die from malaria infection than females, with these differences being most pronounced among TCRbetadelta-/-, microMT, and RAG1 mice. Male mice also exhibited more severe weight loss, anemia, and hypothermia, and higher peak parasitemia than females during infection, with WT, RAG1, TCRbetadelta-/-, and microMT mice exhibiting the most pronounced sexual dimorphism. The absence of IFN-gamma reduced the sex difference in mortality and was more detrimental to females than males. These data suggest that differential transcription and translation of IFN gamma, that is influenced by estrogens, may mediate sex differences in response to malaria. PMID- 16714547 TI - Babesia microti primarily invades mature erythrocytes in mice. AB - Babesia microti is a tick-borne red blood cell parasite that causes babesiosis in people. Its most common vertebrate reservoir is the white-footed mouse. To determine whether B. microti invades reticulocytes, as does the canine pathogen B. gibsoni, we infected the susceptible inbred mouse strains C.B-17.scid and DBA/2 with a clinical isolate of B. microti. Staining of fixed permeabilized red blood cells with 4',6'-diamidino-2-phenylindole or YOYO-1, a sensitive nucleic acid stain, revealed parasite nuclei as large bright dots. Flow cytometric analysis indicated that parasite DNA is primarily found in mature erythrocytes that expressed Babesia antigens but not the transferrin receptor CD71. In contrast, CD71-positive reticulocytes rarely contained Babesia nuclei and failed to express Babesia antigens. Accordingly, the frequency of YOYO-1-positive, CD71 negative cells strongly correlated with parasitemia, defined as the frequency of infected red blood cells assessed on Giemsa-stained blood smears. Importantly, the absolute numbers generated by the two techniques were similar. Parasitemia was modest and transient in DBA/2 mice but intense and sustained in C.B-17.scid mice. In both strains, parasitemia preceded reticulocytosis, but reticulocytes remained refractory to B. microti. In immunocompetent C.B-17 mice, reticulocytosis developed early, despite a marginal and short-lived parasitemia. Likewise, an early reticulocytosis developed in resistant BALB/cBy and B10.D2 mice. These studies establish that B. microti has a tropism for mature erythrocytes. Although reticulocytes are rarely infected, the delayed reticulocytosis in susceptible strains may result from parasite or host activities to limit renewal of the mature erythrocyte pool, thereby preventing an overwhelming parasitemia. PMID- 16714548 TI - Role for dendritic cells in immunoregulation during experimental vaginal candidiasis. AB - Vulvovaginal candidiasis (VVC) caused by the commensal organism Candida albicans remains a significant problem among women of childbearing age, with protection against and susceptibility to infection still poorly understood. While cell mediated immunity by CD4+ Th1-type cells is protective against most forms of mucosal candidiasis, no protective role for adaptive immunity has been identified against VVC. This is postulated to be due to immunoregulation that prohibits a more profound Candida-specific CD4+ T-cell response against infection. The purpose of this study was to examine the role of dendritic cells (DCs) in the induction phase of the immune response as a means to understand the initiation of the immunoregulatory events. Immunostaining of DCs in sectioned murine lymph nodes draining the vagina revealed a profound cellular reorganization with DCs becoming concentrated in the T-cell zone throughout the course of experimental vaginal Candida infection consistent with cell-mediated immune responsiveness. However, analysis of draining lymph node DC subsets revealed a predominance of immunoregulation-associated CD11c+ B220+ plasmacytoid DCs (pDCs) under both uninfected and infected conditions. Staining of vaginal DCs showed the presence of both DEC-205+ and pDCs, with extension of dendrites into the vaginal lumen of infected mice in close contact with Candida. Flow cytometric analysis of draining lymph node DC costimulatory molecules and activation markers from infected mice indicated a lack of upregulation of major histocompatibility complex class II, CD80, CD86, and CD40 during infection, consistent with a tolerizing condition. Together, the results suggest that DCs are involved in the immunoregulatory events manifested during a vaginal Candida infection and potentially through the action of pDCs. PMID- 16714549 TI - Human recombinant antibodies against Plasmodium falciparum merozoite surface protein 3 cloned from peripheral blood leukocytes of individuals with immunity to malaria demonstrate antiparasitic properties. AB - Immunoglobulins from individuals with immunity to malaria have a strong antiparasitic effect when transferred to Plasmodium falciparum malaria infected patients. One prominent target of antiparasitic antibodies is the merozoite surface antigen 3 (MSP-3). We have investigated the antibody response against MSP 3 residues 194 to 257 (MSP-3(194-257)) on the molecular level. mRNA from peripheral blood leukocytes from clinically immune individuals was used as a source of Fab (fragment antibody) genes. A Fab-phage display library was made, and three distinct antibodies designated RAM1, RAM2, and RAM3 were isolated by panning. Immunoglobulin G1 (IgG1) and IgG3 full-length antibodies have been produced in CHO cells. Reactivity with the native parasite protein was demonstrated by immunofluorescence microscopy, flow cytometry, and immunoblotting. Furthermore, the antiparasitic effect of RAM1 has been tested in vitro in an antibody-dependent cellular inhibition (ADCI) assay. Both the IgG1 and the IgG3 versions of the antibody show an inhibitory effect on parasite growth. PMID- 16714550 TI - Eosinophils can function as antigen-presenting cells to induce primary and secondary immune responses to Strongyloides stercoralis. AB - Several studies have demonstrated roles for eosinophils during innate and adaptive immune responses to helminth infections. However, evidence that eosinophils are capable of initiating an immune response to parasite antigens is lacking. The goal of the present in vitro study was to investigate the potential of eosinophils to serve as antigen-presenting cells (APC) and initiate an immune response to parasite antigens. Purified eosinophils were exposed to soluble Strongyloides stercoralis antigens, and the expression of various surface markers involved in cell activation was examined. Antigen-exposed eosinophils showed a sixfold increase in expression levels of CD69 and major histocompatibility complex (MHC) class II, a fourfold increase in levels of T-cell costimulatory molecule CD86, and a twofold decrease in levels of CD62L compared to eosinophils cultured in medium containing granulocyte-macrophage colony-stimulating factor. The ability of eosinophils to present antigen to T cells was determined by culturing them with T cells in vitro. Eosinophils pulsed with antigen stimulated antigen-specific primed T cells and CD4+ T cells to increase interleukin-5 (IL-5) production. The blocking of MHC class II expression on eosinophils inhibited their ability to induce IL-5 production by CD4+ T cells in culture. Antigen pulsed eosinophils were able to prime naive T cells and CD4+ T cells in culture and polarized them into Th2 cells producing IL-5 similar to that induced by antigen-loaded dendritic cells. These results demonstrate that eosinophils are capable of activating antigen-specific Th2 cells inducing the release of cytokines and assist in the priming of naive T cells to initiate Th2 responses against infection. This study highlights the potential of eosinophils to actively induce immune responses against infection by amplifying antigen-specific Th2-cell responses. PMID- 16714551 TI - Interaction of Yersinia pestis with macrophages: limitations in YopJ-dependent apoptosis. AB - The enteropathogenic Yersinia strains are known to downregulate signaling pathways in macrophages by effectors of the type III secretion system, in which YopJ/YopP plays a crucial role. The adverse effects of Yersinia pestis, the causative agent of plague, were examined by infecting J774A.1 cells, RAW264.7 cells, and primary murine macrophages with the EV76 strain and with the fully virulent Kimberley53 strain. Y. pestis exerts YopJ-dependent suppression of tumor necrosis factor alpha secretion and phosphorylation of mitogen-activated protein kinases and thus resembles enteropathogenic Yersinia. However, Y. pestis is less able to activate caspases, to suppress NF-kappaB activation, and to induce apoptosis in macrophages than the high-virulence Y. enterocolitica WA O:8 strain. These differences appear to be related to lower efficiency of YopJ effector translocation by Y. pestis. The efficiencies of effector translocation and of apoptosis induction can be enhanced either by using a high bacterial load in a synchronized infection or by overexpressing exogenous YopJ in Y. pestis. Replacing YopJ with the homologous Y. enterocolitica effector YopP can further enhance these effects. Overexpression of YopP in a yopJ-deleted Y. pestis background leads to rapid and effective translocation into target cells, providing Y. pestis with the high cytotoxic potential of Y. enterocolitica WA O:8. We suggest that the relative inferiority of Y. pestis in triggering cell death in macrophages may be advantageous for its in vivo propagation in the early stages of infection. PMID- 16714552 TI - Environmental signals generate a differential and coordinated expression of the heme receptor gene family of Bartonella quintana. AB - Of all bacteria, Bartonella quintana has the highest reported in vitro hemin requirement, yet an explanation for this remains elusive. To produce diseases such as trench fever, endocarditis, and bacillary angiomatosis, B. quintana must survive and replicate in the disparate environments of the Pediculus humanus corporis (body louse) gut and the human vasculature. We previously identified a five-member family of hemin binding proteins (Hbps) synthesized by B. quintana that bind hemin on the outer surface but share no similarity to known bacterial heme receptors. In the present study, we examine the transcription, regulation, and synthesis of this virulence factor family by cultivation of the bacterium in environments that simulate natural heme, oxygen, and temperature conditions encountered in the host and insect vector. First, quantitative real-time PCR data show that hbpC expression is regulated by temperature, where a >100-fold increase in transcript quantity was seen at 30 degrees C relative to 37 degrees C, suggesting that HbpC synthesis would be greatest in the cooler temperature of the louse. Second, cultivation at human bloodstream oxygen concentration (5% relative to 21% atmospheric) significantly decreases the transcript quantity of all hbp genes, indicating that expression is influenced by O2 and/or reactive oxygen species. Third, a differential expression pattern within the hbp family is revealed when B. quintana is grown in a range of hemin concentrations: subgroup I (hbpC and hbpB) predominates in a simulated louse environment (high heme), and subgroup II (hbpA, hbpD, and hbpE) is preferentially expressed in a simulated human background (low heme). By using two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis, immunoblotting, and matrix-assisted laser desorption ionization-time of flight mass spectrometry fingerprinting, we demonstrate that synthesis of HbpA correlates with hbpA transcript increases observed at low hemin concentrations. Finally, an hbpA promoter-lacZ reporter construct in B. quintana demonstrates that a transcriptional regulator(s) is controlling the expression of hbpA through a cis-acting regulatory element located in the hbpA promoter region. PMID- 16714553 TI - Plasmodium falciparum-infected erythrocytes increase intercellular adhesion molecule 1 expression on brain endothelium through NF-kappaB. AB - Sequestration of Plasmodium falciparum-infected erythrocytes (Pf-IRBC) in postcapillary brain endothelium is a hallmark of cerebral malaria (CM) pathogenesis. There is a correlation between adherent Pf-IRBC and increased expression of intercellular cell adhesion molecule 1 (ICAM-1), which is also a receptor for Pf-IRBC on human brain microvascular endothelial cells (HBMEC). The underlying mechanism for the increased ICAM-1 expression has not been clearly defined. Therefore, we investigated the mechanisms of ICAM-1 expression on isolated HBMEC after exposure to Pf-IRBC. Ultrastructural characterization of the model confirmed that there was attachment through both Pf-IRBC knobs and HBMEC microvillus protrusions. Pf-IRBC induced a dose- and time-dependent increase in ICAM-1 expression on HBMEC that was specific for human brain endothelium and was not observed with human umbilical vein endothelium. Involvement of both membrane associated Pf-IRBC proteins and parasite-derived soluble factors with the increase in ICAM-1 expression was demonstrated by surface trypsinization and fractionation. Pf-IRBC exposure induced nuclear translocation of NF-kappaB in HBMEC, which was linked to ICAM-1 expression, as shown by use of specific inhibitors of the transcription factor NF-kappaB and immunocytochemistry. In addition, inhibition of reactive oxygen species decreased Pf-IRBC-induced ICAM-1 expression on HBMEC. Parasite-induced ICAM-1 expression explains the localization of this molecule on brain endothelium in postmortem CM brain samples. By increasing ICAM-1 expression, Pf-IRBC may increase their sequestration, thereby perpetuating CM. PMID- 16714554 TI - Differential expression of the Bhmp39 major outer membrane proteins of Brachyspira hyodysenteriae. AB - The enteric, anaerobic spirochete Brachyspira hyodysenteriae is the causative agent of swine dysentery, a severe mucohemorrhagic diarrheal disease of pigs that has economic significance in every major pork-producing country. Recent investigation into potential vaccine candidates has focused on the outer membrane proteins of B. hyodysenteriae. Bhmp39 (formerly Vsp39) is the most abundant surface-exposed outer membrane protein of B. hyodysenteriae; its predicted gene sequence has previously been shown to share sequence similarity to eight genes divided evenly between two paralogous loci. The peptide sequence suggested that Bhmp39 is encoded by one of these genes, bhmp39h. The biological significance of maintaining eight homologous bhmp39 genes is unclear, though it has been proposed that this may play a role in antigenic variation. In this study, real-time, reverse transcription-PCR was used to demonstrate that bhmp39f and bhmp39h were the transcripts most abundantly expressed by B. hyodysenteriae strain B204 cultured under in vitro growth conditions. Mass spectrometry data of the purified 39-kDa membrane protein showed that both Bhmp39f and Bhmp39h were present. Northern blot analysis across predicted Rho-independent terminators demonstrated that the genes of the bhmp39efgh locus result in monocistronic transcripts. PMID- 16714555 TI - Basis for the failure of Francisella tularensis lipopolysaccharide to prime human polymorphonuclear leukocytes. AB - Francisella tularensis is the intracellular gram-negative coccobacillus that causes tularemia, and its virulence and infectiousness make it a potential agent of bioterrorism. Previous studies using mononuclear leukocytes have shown that the lipopolysaccharide (LPS) of F. tularensis is neither a typical proinflammatory endotoxin nor an endotoxin antagonist. This inertness suggests that F. tularensis LPS does not bind host LPS-sensing molecules such as LPS binding protein (LBP). Using priming of the polymorphonuclear leukocyte (PMN) oxidase as a measure of endotoxicity, we found that F. tularensis live vaccine strain LPS did not behave like either a classic endotoxin or an endotoxin antagonist in human PMNs, even when the concentration of LBP was limiting. Furthermore, F. tularensis LPS did not compete with a radiolabeled lipooligosaccharide from Neisseria meningitidis for binding to LBP or to the closely related PMN granule protein, bactericidal/permeability-increasing protein. Our results suggest that the inertness of F. tularensis LPS and the resistance of F. tularensis to oxygen-independent PMN killing may result from the inability of F. tularensis LPS to be recognized by these important LPS-sensing molecules of the innate immune system. PMID- 16714556 TI - Membrane vesicles shed by Legionella pneumophila inhibit fusion of phagosomes with lysosomes. AB - When cultured in broth to the transmissive phase, Legionella pneumophila infects macrophages by inhibiting phagosome maturation, whereas replicative-phase cells are transported to the lysosomes. Here we report that the ability of L. pneumophila to inhibit phagosome-lysosome fusion correlated with developmentally regulated modifications of the pathogen's surface, as judged by its lipopolysaccharide profile and by its binding to a sialic acid-specific lectin and to the hydrocarbon hexadecane. Likewise, the composition of membrane vesicles shed by L. pneumophila was developmentally regulated, based on binding to the lectin and to the lipopolysaccharide-specific monoclonal antibody 3/1. Membrane vesicles were sufficient to inhibit phagosome-lysosome fusion by a mechanism independent of type IV secretion, since only approximately 25% of beads suspended with or coated by vesicles from transmissive phase wild type or dotA secretion mutants colocalized with lysosomal probes, whereas approximately 75% of beads were lysosomal when untreated or presented with vesicles from the L. pneumophila letA regulatory mutant or E. coli. As observed previously for L. pneumophila infection of mouse macrophages, vesicles inhibited phagosome-lysosome fusion only temporarily; by 10 h after treatment with vesicles, macrophages delivered approximately 72% of ingested beads to lysosomes. Accordingly, in the context of the epidemiology of the pneumonia Legionnaires' disease and virulence mechanisms of Leishmania and Mycobacteria, we discuss a model here in which L. pneumophila developmentally regulates its surface composition and releases vesicles into phagosomes that inhibit their fusion with lysosomes. PMID- 16714557 TI - Infection of human dendritic cells with a Mycobacterium tuberculosis sigE mutant stimulates production of high levels of interleukin-10 but low levels of CXCL10: impact on the T-cell response. AB - The Mycobacterium tuberculosis genome encodes 13 sigma factors. We have previously shown that mutations in some of these transcriptional activators render M. tuberculosis sensitive to various environmental stresses and can attenuate the virulence phenotype. In this work, we focused on extracytoplasmic factor sigmaE and studied the effects induced by the deletion of its structural gene (sigE) in the infection of human monocyte-derived dendritic cells (MDDC). We found that the wild-type M. tuberculosis strain (H37Rv), the sigE mutant (ST28), and the complemented strain (ST29) were able to infect dendritic cells (DC) to similar extents, although at 4 days postinfection a reduced ability to grow inside MDDC was observed for the sigE mutant ST28. After mycobacterium capture, the majority of MDDC underwent full maturation and expressed both inflammatory cytokines, such as tumor necrosis factor alpha, and the regulatory cytokines interleukin-12 (IL-12), IL-18, and beta interferon (IFN-beta). Conversely, a higher level of production of IL-10 was observed in ST28-infected MDDC compared to H37Rv- or ST29-infected cell results. However, in spite of the presence of IL 10, supernatants from ST28-infected DC induced IFN-gamma production by T cells similarly to those from H37Rv-infected DC culture. On the other hand, IL-10 impaired CXCL10 production in sigE mutant-infected DC and, indeed, its neutralization restored CXCL10 secretion. In line with these results, supernatants from ST28-infected cells showed a decreased capability to recruit CXCR3+ CD4+ T cells compared to those obtained from H37Rv-infected DC culture. Thus, our findings suggest that the sigE mutant-induced secretion of IL-10 inhibits CXCL10 expression and, in turn, the recruitment of activated-effector cells involved in the formation of granulomas. PMID- 16714558 TI - Borrelia burgdorferi lacking BBK32, a fibronectin-binding protein, retains full pathogenicity. AB - BBK32, a fibronectin-binding protein of Borrelia burgdorferi, is one of many surface lipoproteins that are differentially expressed by the Lyme disease spirochete at various stages of its life cycle. The level of BBK32 expression in B. burgdorferi is highest during infection of the mammalian host and lowest in flat ticks. This temporal expression profile, along with its fibronectin-binding activity, strongly suggests that BBK32 may play an important role in Lyme pathogenesis in the host. To test this hypothesis, we constructed an isogenic BBK32 deletion mutant from wild-type B. burgdorferi B31 by replacing the BBK32 gene with a kanamycin resistance cassette through homologous recombination. We examined both the wild-type strain and the BBK32 deletion mutant extensively in the experimental mouse-tick model of the Borrelia life cycle. Our data indicated that B. burgdorferi lacking BBK32 retained full pathogenicity in mice, regardless of whether mice were infected artificially by syringe inoculation or naturally by tick bite. The loss of BBK32 expression in the mutant had no adverse effect on spirochete acquisition (mouse-to-tick) and transmission (tick-to-mouse) processes. These results suggest that additional B. burgdorferi proteins can complement the function of BBK32, fibronectin binding or otherwise, during the natural spirochete life cycle. PMID- 16714559 TI - Th1 cytokines facilitate CD8-T-cell-mediated early resistance to infection with Mycobacterium tuberculosis in old mice. AB - Numerous immunological defects begin to emerge as an individual ages, the consequence of which is heightened susceptibility to infectious diseases. Despite this decline in immune function, old mice display an early transient resistance to Mycobacterium tuberculosis infection in the lung, which is dependent on CD8 T cells and gamma interferon (IFN-gamma) production. In this study, we investigated the mechanism of resistance by examining the CD8-T-cell phenotype and function in old naive and M. tuberculosis-infected mice. Pulmonary CD8 T cells from naive old mice expressed cell surface markers of memory in addition to receptors for several Th1 cytokines. Stimulation of lung cells from naive old mice with a combination of Th1 cytokines (interleukin-2 [IL-2], IL-12, and IL-18) resulted in nonspecific production of IFN-gamma by memory CD8 T cells. Following aerosol infection with M. tuberculosis, the lungs of old mice contained significantly more IL-12, IL-18, and IFN-gamma than the lungs of young mice contained. Together, these data demonstrate that the increased and early production of Th1 cytokines in the lungs of M. tuberculosis-infected old mice, in combination with CD8 T cells that can nonspecifically produce IFN-gamma, leads to transient control of M. tuberculosis growth in the lungs of old mice. Further characterization of this mechanism should provide essential information regarding the aging immune system and should contribute to the development of novel strategies to decrease the morbidity and mortality of the aging population associated with infectious diseases. PMID- 16714560 TI - MyD88-dependent responses involving toll-like receptor 2 are important for protection and clearance of Legionella pneumophila in a mouse model of Legionnaires' disease. AB - Legionella pneumophila is a gram-negative facultative intracellular parasite of macrophages. Although L. pneumophila is the causative agent of a severe pneumonia known as Legionnaires' disease, it is likely that most infections caused by this organism are cleared by the host innate immune system. It is predicted that host pattern recognition proteins belonging to the Toll-like receptor (TLR) family are involved in the protective innate immune responses. We examined the role of TLR mediated responses in L. pneumophila detection and clearance using genetically altered mouse hosts in which the macrophages are permissive for L. pneumophila intracellular replication. Our data demonstrate that cytokine production by bone marrow-derived macrophages (BMMs) in response to L. pneumophila infection requires the TLR adapter protein MyD88 and is reduced in the absence of TLR2 but not in the absence of TLR4. Bacterial growth ex vivo in BMMs from MyD88-deficient mice was not enhanced compared to bacterial growth ex vivo in BMMs from heterozygous littermate controls. Wild-type mice were able to clear L. pneumophila from the lung, whereas respiratory infection of MyD88-deficient mice caused death that resulted from robust bacterial replication and dissemination. In contrast to an infection with virulent L. pneumophila, MyD88-deficient mice were able to clear infections with L. pneumophila dotA mutants, indicating that MyD88-independent responses in the lung are sufficient to clear bacteria that are unable to replicate intracellularly. In vivo growth of L. pneumophila was enhanced in the lungs of TLR2-deficient mice, which resulted in a delay in bacterial clearance. No significant differences were observed in the growth and clearance of L. pneumophila in the lungs of TLR4-deficient mice and heterozygous littermate control mice. Our data indicate that MyD88 is crucial for eliciting a protective innate immune response against virulent L. pneumophila and that TLR2 is one of the pattern recognition receptors involved in initiating this MyD88 dependent response. PMID- 16714561 TI - Transcriptional induction of the Pseudomonas aeruginosa type III secretion system by low Ca2+ and host cell contact proceeds through two distinct signaling pathways. AB - The opportunistic pathogen Pseudomonas aeruginosa utilizes a type III secretion system (T3SS) to intoxicate eukaryotic host cells. Transcription of the T3SS is induced under calcium-limited growth conditions or following intimate contact of P. aeruginosa with host cells. In the present study, we demonstrate that expression of the T3SS is controlled by two distinct regulatory mechanisms and that these mechanisms are differentially activated in a host cell-dependent manner. The first mechanism is dependent upon ExsC, a regulatory protein that couples transcription of the T3SS to the activity of the type III secretion machinery. ExsC is essential for induction of the T3SS under low-calcium-growth conditions and for T3SS-dependent cytotoxicity towards social amoebae, insect cells, and erythrocytes. The second regulatory mechanism functions independently of ExsC and is sufficient to elicit T3SS-dependent cytotoxicity towards certain types of mammalian cells. Although this second pathway (ExsC independent) is sufficient, an exsC mutant demonstrates a lag in the induction of cytotoxicity towards Chinese hamster ovary cells and is attenuated for virulence in a mouse pneumonia model. We propose that the ExsC-dependent pathway is required for full cytotoxicity towards all host cell types tested whereas the ExsC-independent pathway may represent an adaptation that allows P. aeruginosa to increase expression of the T3SS in response to specific types of mammalian cells. PMID- 16714562 TI - Bile acids enhance invasiveness of Cryptosporidium spp. into cultured cells. AB - Bile salts such as sodium taurocholate (NaTC) are routinely used to induce the excystation of Cryptosporidium oocysts. Here we show that NaTC significantly enhanced the invasion of several cultured cell lines by freshly excysted Cryptosporidium parvum and Cryptosporidium hominis sporozoites. A variety of purified bile salts or total bile from bovine also enhanced the invasion of cultured cells by C. parvum. Further studies demonstrated that NaTC increased protein secretion and gliding motility of sporozoites, the key processes for successful invasion. These observations may lead to improved Cryptosporidium infectivity of cultured cells and help future studies on the host-parasite interaction. PMID- 16714563 TI - Identification of a novel antigen of Schistosoma mansoni shared with Plasmodium falciparum and evaluation of different cross-reactive antibody subclasses induced by human schistosomiasis and malaria. AB - Plasmodium falciparum and Schistosoma mansoni are often found in human coinfections, and cross-reactive antibodies to different components of the two parasites have been detected. In this work, we identified a cross-reactive S. mansoni gene product, referred to as SmLRR, that seems to belong to the leucine rich repeat protein family. Comparative analysis of SmLRR revealed 57% similarity with a putative gene product encoded in the P. falciparum genome. Antibodies to SmLRR were found in experimental infections and in both S. mansoni- and P. falciparum-infected individuals. Correlative analysis of human anti-SmLRR responses in Kenya and Uganda suggested that malaria and schistosomiasis drive the immunoglobulin G3 (IgG3) and IgG4 isotypes, respectively, against SmLRR, suggesting that there is differential regulation of cross-reactive isotypes depending on the infection. In addition, the levels of anti-SmLRR IgG4, but not the levels of IgG3, correlated positively with the intensity of S. mansoni infection. PMID- 16714564 TI - Elevated plasma phenylalanine in severe malaria and implications for pathophysiology of neurological complications. AB - Cerebral malaria is associated with decreased production of nitric oxide and decreased levels of its precursor, l-arginine. Abnormal amino acid metabolism may thus be an important factor in malaria pathogenesis. We sought to determine if other amino acid abnormalities are associated with disease severity in falciparum malaria. Subjects were enrolled in Dar es Salaam, Tanzania (children) (n = 126), and Papua, Indonesia (adults) (n = 156), in two separate studies. Plasma samples were collected from subjects with WHO-defined cerebral malaria (children), all forms of severe malaria (adults), and uncomplicated malaria (children and adults). Healthy children and adults without fever or illness served as controls. Plasma amino acids were measured using reverse-phase high-performance liquid chromatography with fluorescence detection. Several plasma amino acids were significantly lower in the clinical malaria groups than in healthy controls. Despite the differences, phenylalanine was the only amino acid with mean levels outside the normal range (40 to 84 microM) and was markedly elevated in children with cerebral malaria (median [95% confidence interval], 163 [134 to 193] microM; P < 0.0001) and adults with all forms of severe malaria (median [95% confidence interval], 129 [111 to 155] microM; P < 0.0001). In adults who survived severe malaria, phenylalanine levels returned to normal, with clinical improvement (P = 0.0002). Maintenance of plasma phenylalanine homeostasis is disrupted in severe malaria, leading to significant hyperphenylalaninemia. This is likely a result of an acquired abnormality in the function of the liver enzyme phenylalanine hydroxylase. Determination of the mechanism of this abnormality may contribute to the understanding of neurological complications in malaria. PMID- 16714565 TI - Variation in the presence of neuraminidase genes among Streptococcus pneumoniae isolates with identical sequence types. AB - Streptococcus pneumoniae frequently colonizes the upper respiratory tract of young children and is an important cause of otitis media and invasive disease. Carriage is more common than disease, yet the genetic factors that predispose a given clone for disease are not known. The relationship between capsule type, genetic background, and virulence is complex, and important questions remain regarding how pneumococcal clones differ in their ability to cause disease. Pneumococcal neuraminidase cleaves sialic acid-containing substrates and is thought to be important for pneumococcal virulence. We describe the distribution of multilocus sequence types (ST), capsule type, and neuraminidase genes among 342 carriage, middle ear, blood, and cerebrospinal fluid (CSF) pneumococcal strains from young children. We found 149 STs among our S. pneumoniae isolates. nanA was present in all strains, while nanB and nanC were present in 96% and 51% of isolates, respectively. The distribution of nanC varied among the strain collections from different tissue sources (P = 0.03). The prevalence of nanC was 1.41 (95% confidence interval, 1.11, 1.79) times higher among CSF isolates than among carriage isolates. We identified isolates of the same ST that differed in the presence of nanB and nanC. These studies demonstrate that virulence determinants, other than capsule loci, vary among strains of identical ST. Our studies suggest that the presence of nanC may be important for tissue-specific virulence. Studies that both incorporate MLST and take into account additional virulence determinants will provide a greater understanding of the pneumococcal virulence potential. PMID- 16714566 TI - Differential genome contents of nontypeable Haemophilus influenzae strains from adults with chronic obstructive pulmonary disease. AB - Haemophilus influenzae is an important cause of otitis media in children and lower respiratory infection in adults with chronic obstructive pulmonary disease (COPD). Patients with COPD experience periodic exacerbations that are associated with acquisition of new bacterial strains. However, not every strain acquisition is associated with exacerbation. To test the hypothesis that genetic differences among strains account for differences in pathogenic potential, a microarray consisting of 4,992 random 1.5- to 3-kb genomic fragments of an exacerbation strain was constructed. Competitive hybridization was performed using six strains associated with exacerbation as well as five strains associated with asymptomatic colonization. Seven sequences that were absent in all five colonization strains and present in at least two exacerbation strains were identified. One such sequence was a previously unreported gene with high homology to the meningococcal immunoglobulin A (IgA) protease gene, which is distinct from the previously described H. influenzae IgA protease. To assess the distribution of the seven sequences among well-characterized strains of H. influenzae, 59 exacerbation strains and 73 asymptomatic colonization strains were screened by PCR for the presence of these sequences. The presence or absence of any single sequence was not significantly associated with exacerbations of COPD. However, logistic regression and subgroup analysis identified combinations of the presence and absence of genes that are associated with exacerbations. These results indicate that patterns of genes are associated with the ability of strains of H. influenzae to cause exacerbations of COPD, supporting the concept that differences in pathogenic potential are based in part on genomic differences among infecting strains, not merely host factors. PMID- 16714567 TI - Analyses of Five gallinacin genes and the Salmonella enterica serovar Enteritidis response in poultry. AB - Gallinacins in poultry are functional equivalents of mammalian beta-defensins, which constitute an integral component of the innate immune system. Salmonella enterica serovar Enteritidis is a gram-negative bacterium that negatively affects both human and animal health. To analyze the association of genetic variations of the gallinacin genes with the phenotypic response to S. enterica serovar Enteritidis, an F1 population of chickens was created by crossing four outbred broiler sires to dams of two highly inbred lines. The F1 chicks were evaluated for bacterial colonization after pathogenic S. enterica serovar Enteritidis inoculation and for circulating antibody levels after inoculation with S. enterica serovar Enteritidis bacterin vaccine. Five candidate genes were studied, including gallinacins 2, 3, 4, 5, and 7. Gene fragments were sequenced from the founder individuals of the resource population, and a mean of 13.2 single nucleotide polymorphisms (SNP) per kilobase was identified. One allele-defining SNP per gene was utilized to test for statistical associations of sire alleles with progeny response to S. enterica serovar Enteritidis. Among the five gallinacin genes evaluated, the Gal3 and Gal7 SNPs in broiler sires were found to be associated with antibody production after S. enterica serovar Enteritidis vaccination. Utilization of these SNPs as molecular markers for the response to S. enterica serovar Enteritidis may result in the enhancement of the immune response in poultry. PMID- 16714568 TI - Gamma interferon, tumor necrosis factor alpha, and nitric oxide synthase 2, key elements of cellular immunity, perform critical protective functions during humoral defense against lethal pulmonary Yersinia pestis infection. AB - Pulmonary infection by Yersinia pestis causes pneumonic plague, a rapidly progressing and often fatal disease. To aid the development of safe and effective pneumonic plague vaccines, we are deciphering mechanisms used by the immune system to protect against lethal pulmonary Y. pestis infection. In murine pneumonic plague models, passive transfer of convalescent-phase sera confers protection, as does active vaccination with live Y. pestis. Here, we demonstrate that protection by either protocol relies upon both gamma interferon (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) cytokines classically associated with type 1 cellular immunity. In both protocols, abrogating IFN-gamma or TNF-alpha activity significantly decreases survival and increases the bacterial burden in pulmonary, splenic, and hepatic tissues. Neutralization of either cytokine also counteracts challenge-induced, vaccination-dependent upregulation of nitric oxide synthase 2 (NOS2). Moreover, genetic depletion of NOS2 suppresses protection conferred by serotherapy. We conclude that IFN-gamma, TNF-alpha, and NOS2, key elements of cellular immunity, perform critical protective functions during humoral defense against lethal pulmonary Y. pestis challenge. These observations strongly suggest that plague vaccines should strive to maximally prime both cellular and humoral immunity. PMID- 16714569 TI - High levels of interleukin-10 impair resistance to pulmonary coccidioidomycosis in mice in part through control of nitric oxide synthase 2 expression. AB - We have shown previously that there is a direct correlation between IL-10 levels and susceptibility to Coccidioides immitis peritonitis in C57BL/6 (B6), DBA/2, and BXD recombinant inbred mice. We now show that B6 mice are also more susceptible to C. immitis pneumonia and that interleukin-10 (IL-10)-deficient (IL 10-/-) B6 mice are more resistant to C. immitis pneumonia. In addition, we established that high levels of IL-10 are sufficient to make genetically resistant mice susceptible to both C. immitis peritonitis and pneumonia by infecting h.IL-10 transgenic mice. Infected h.IL-10 transgenic mice express lower levels of gamma interferon, IL-12 p40, and inducible nitric oxide synthetase 2 (NOS2) mRNA in their lungs, implicating inducible NOS as a defense mechanism in this disease. We treated DBA/2 mice with aminoguanidine, and they became more susceptible to C. immitis peritonitis and pneumonia. We conclude that high levels of IL-10 are both necessary and sufficient to make mice susceptible to C. immitis, regardless of the genetic background of the mice, and that IL-10 impairs resistance to C. immitis in part by suppressing NO synthesis. PMID- 16714570 TI - High frequency of CD4+ T cells specific for the TB10.4 protein correlates with protection against Mycobacterium tuberculosis infection. AB - TB10.4 is a newly identified antigen of Mycobacterium tuberculosis recognized by human and murine T cells upon mycobacterial infection. Here, we show that immunization with Mycobacterium bovis BCG induces a strong, genetically controlled, Th1 immune response against TB10.4 in mice. BALB/c and C57BL/6 strains behave as high and low responders to TB10.4 protein, respectively. The TB10.4:74-88 peptide was identified as an immunodominant CD4+ T-cell epitope for H-2d mice. Since recent results, as well as the present study, have raised interest in TB10.4 as a subunit vaccine, we analyzed immune responses induced by this antigen delivered by a new vector, the adenylate cyclase (CyaA) of Bordetella pertussis. CyaA is able to target dendritic cells and to deliver CD4+ or CD8+ T-cell epitopes to the major histocompatibility complex class II/I molecule presentation pathways, triggering specific Th1 or cytotoxic T-lymphocyte (CTL) responses. Several CyaA harboring either the entire TB10.4 protein or various subfragments containing the TB10.4:20-28 CTL epitope were shown to induce TB10.4-specific Th1 CD4+ and CD8+ T-cell responses. However, none of the recombinant CyaA, injected in the absence of adjuvant, was able to induce protection against M. tuberculosis infection. In contrast, TB10.4 protein administered with a cocktail of strong adjuvants that triggered a strong Th1 CD4+ T-cell response induced significant protection against M. tuberculosis challenge. These results confirm the potential value of the TB10.4 protein as a candidate vaccine and show that the presence of high frequencies of CD4+ T cells specific to this strong immunogen correlates with protection against M. tuberculosis infection. PMID- 16714571 TI - Heterogeneity in tandem octanucleotides within Haemophilus influenzae lipopolysaccharide biosynthetic gene losA affects serum resistance. AB - Haemophilus influenzae is subject to phase variation mediated by changes in the length of simple sequence repeat regions within several genes, most of which encode either surface proteins or enzymes involved in the synthesis of lipopolysaccharides (LPS). The translational repeat regions that have been described thus far all consist of tandemly repeated tetranucleotides. We describe an octanucleotide repeat region within a putative LPS biosynthetic gene, losA. Approximately 20 percent of nontypeable H. influenzae strains contain copies of losA and losB in a genetic locus flanked by infA and ksgA. Of 30 strains containing losA at this site, 24 contained 2 tandem copies of the octanucleotide CGAGCATA, allowing full-length translation of losA (on), and 6 strains contained 3, 4, 6, or 10 tandem copies (losA off). For a serum-sensitive strain, R3063, with losA off (10 repeat units), selection for serum-resistant variants yielded a heterogeneous population in which colonies with increased serum resistance had losA on (2, 8, or 11 repeat units), and colonies with unchanged sensitivity to serum had 10 repeats. Inactivation of losA in strains R3063 and R2846 (strain 12) by insertion of the cat gene decreased the serum resistance of these strains compared to losA-on variants and altered the electrophoretic mobility of LPS. We conclude that expression of losA, a gene that contributes to LPS structure and affects serum resistance, is determined by octanucleotide repeat variation. PMID- 16714572 TI - Identification of Staphylococcus aureus proteins recognized by the antibody mediated immune response to a biofilm infection. AB - Staphylococcus aureus causes persistent, recurrent infections (e.g., osteomyelitis) by forming biofilms. To survey the antibody-mediated immune response and identify those proteins that are immunogenic in an S. aureus biofilm infection, the tibias of rabbits were infected with methicillin-resistant S. aureus to produce chronic osteomyelitis. Sera were collected prior to infection and at 14, 28, and 42 days postinfection. The sera were used to perform Western blot assays on total protein from biofilm grown in vitro and separated by two dimensional gel electrophoresis. Those proteins recognized by host antibodies in the harvested sera were identified via matrix-assisted laser desorption ionization-time of flight analysis. Using protein from mechanically disrupted total and fractionated biofilm protein samples, we identified 26 and 22 immunogens, respectively. These included a cell surface-associated beta lactamase, lipoprotein, lipase, autolysin, and an ABC transporter lipoprotein. Studies were also performed using microarray analyses and confirmed the biofilm specific up-regulation of most of these genes. Therefore, although the biofilm antigens are recognized by the immune system, the biofilm infection can persist. However, these proteins, when delivered as vaccines, may be important in directing the immune system toward an early and effective antibody-mediated response to prevent chronic S. aureus infections. Previous works have identified S. aureus proteins that are immunogenic during acute infections, such as sepsis. However, this is the first work to identify these immunogens during chronic S. aureus biofilm infections and to simultaneously show the global relationship between the antigens expressed during an in vivo infection and the corresponding in vitro transcriptomic and proteomic gene expression levels. PMID- 16714573 TI - Iha from an Escherichia coli urinary tract infection outbreak clonal group A strain is expressed in vivo in the mouse urinary tract and functions as a catecholate siderophore receptor. AB - Virulence factors of pathogenic Escherichia coli belonging to a recently emerged and disseminated clonal group associated with urinary tract infection (UTI), provisionally designated clonal group A (CGA), have not been experimentally investigated. We used a mouse model of ascending UTI with CGA member strain UCB34 in order to identify genes of CGA that contribute to UTI. iha was identified to be expressed by strain UCB34 in the mouse kidney using selective capture of transcribed sequences. iha from strain UCB34 demonstrated a siderophore receptor phenotype when cloned in a catecholate siderophore receptor-negative E. coli K-12 strain, as shown by growth promotion experiments and uptake of (55)Fe complexed to enterobactin or its linear 2, 3-dihydroxybenzoylserine (DHBS) siderophore derivatives. Siderophore-mediated growth promotion by Iha was TonB dependent. Growth and iron uptake were more marked with linear DHBS derivatives than with purified enterobactin. The reported phenotype of adherence to epithelial cells conferred by expressing iha from a multicopy cloning vector in a poorly adherent E. coli K-12 host strain was confirmed to be specific to iha, in comparison with other siderophore receptor genes. iha expression was regulated by the ferric uptake regulator Fur and by iron availability, as shown by real-time reverse transcriptase PCR. In a competitive infection experiment using the mouse UTI model, wild-type strain UCB34 significantly outcompeted an isogenic iha null mutant. Iha thus represents a Fur-regulated catecholate siderophore receptor that, uniquely, exhibits an adherence-enhancing phenotype and is the first described urovirulence factor identified in a CGA strain. PMID- 16714575 TI - Genetic variation among isolates of Sarcocystis neurona, the agent of protozoal myeloencephalitis, as revealed by amplified fragment length polymorphism markers. AB - Sarcocystis neurona causes serious neurological disease in horses and other vertebrates in the Americas. Based on epidemiological data, this parasite has recently emerged. Here, the genetic diversity of Sarcocystis neurona was evaluated using the amplified fragment length polymorphism (AFLP) method. Fifteen S. neurona taxa from different regions collected over the last 10 years were used; six isolates were from clinically diseased horses, eight isolates were from wild-caught opossums (Didelphis virginiana), and one isolate was from a cowbird (Molothrus ater). Additionally, four outgroup taxa were also fingerprinted. Nine primer pairs were used to generate AFLP patterns, with a total number of amplified fragments ranging from 30 to 60, depending on the isolate and primers tested. Based on the presence/absence of amplified AFLP fragments and pairwise similarity values, all the S. neurona isolates tested were clustered in one monophyletic group. No significant correlation could be found between genomic similarity and host origin of the S. neurona isolates. AFLP revealed significant intraspecific genetic variations, and S. neurona appeared as a highly variable species. Furthermore, linkage disequilibrium analysis suggested that S. neurona populations within Michigan have an intermediate type of population structure that includes characteristics of both clonal and panamictic population structures. AFLP is a reliable molecular technique that has provided one of the most informative approaches to ascertain phylogenetic relationships in S. neurona and its closest relatives, allowing them to be clustered by relative similarity using band matching and unweighted pair group method with arithmetic mean analysis, which may be applicable to other related protozoal species. PMID- 16714574 TI - Flagellin-independent regulation of chemokine host defense in Campylobacter jejuni-infected intestinal epithelium. AB - Campylobacter jejuni is a leading cause of bacterial food-borne diarrheal disease throughout the world and the most frequent antecedent of autoimmune neuropathy Guillain-Barre syndrome. While infection is associated with immune memory, little is known regarding the role of the epithelium in targeting dendritic cells (DC) for initiating the appropriate adaptive immune response to C. jejuni. The objective of this study was to define the role for the intestinal epithelium in the induction of the adaptive immune response in C. jejuni infection by assessing the production of DC and T-cell chemoattractants. Human T84 epithelial cells were used as model intestinal epithelia. Infection of T84 cells with C. jejuni dose- and time-dependently up-regulated DC and T-cell chemokine gene transcription and secretion. Induction required live bacteria and was in the physiologically relevant direction for attraction of mucosal immunocytes. C. jejuni-activated NF kappaB signaling was shown to be essential for proinflammatory chemokine secretion. Notably, C. jejuni secretion occurred independently of flagellin identification by Toll-like receptor 5. Secretion of a DC chemoattractant by differing clinical C. jejuni isolates suggested adherence/invasion were key virulence determinants of epithelial chemokine secretion. The regulated epithelial expression of DC and T-cell chemoattractants suggests a mechanism for the directed trafficking of immune cells required for the initiation of adaptive immunity in campylobacteriosis. Chemokine secretion occurs despite Campylobacter evasion of the flagellin pattern recognition receptor, suggesting that alternate host defense strategies limit disease pathogenesis. PMID- 16714577 TI - Characterization of a novel high-affinity monoclonal immunoglobulin G antibody against the ricin B subunit. AB - There is an urgent need for the development of a passive immunotherapy against the category B select agent ricin, a lethal ribosome-inactivating toxin composed of an enzymatic A subunit (RTA) and a single binding B subunit (RTB). To date, only one monoclonal antibody (MAb), a mouse immunoglobulin G (IgG1) against RTA called R70, has been deemed sufficiently potent in animal models to warrant further testing in humans. In this study, we have identified and characterized MAb 24B11, a murine IgG1 directed against RTB. In a Vero cell cytotoxicity assay, 24B11 was approximately two times more effective at neutralizing ricin than was R70. The equilibrium dissociation constants of 24B11 (KD = 4.2 x 10(-9) M) and R70 (KD = 3.2 x 10(-9) M) were virtually identical, suggesting that the difference in neutralization activity between the two MAbs was not due to differing affinities for the toxin. 24B11 blocked ricin attachment to galactoside receptors on primary mouse splenocytes and on the apical surfaces of human mucosal epithelial cell monolayers. Surprisingly, R70 also effectively interfered with ricin attachment to receptors on cell surfaces. Using a phage-displayed peptide library, we determined that 24B11 binds an epitope on RTB adjacent to, but not within, one of the two galactose binding domains. Finally, we demonstrate that R70 and 24B11, when combined, function synergistically to neutralize ricin in vitro, raising the possibility that these two MAbs could serve as a novel immunotherapeutic in vivo. PMID- 16714576 TI - Immunoglobulin A antibodies against ricin A and B subunits protect epithelial cells from ricin intoxication. AB - Epithelial cells of the respiratory and gastrointestinal tracts are extremely vulnerable to the cytotoxic effects of ricin, a Shiga-like toxin with ribosome inactivating properties. While mucosal immunity to ricin correlates with secretory immunoglobulin A (IgA) antibody levels in vivo, the potential of IgA to protect epithelial cells from ricin in vitro has not been examined due to the unavailability of well-defined antitoxin IgA antibodies. Here we report the characterization of four monoclonal IgA antibodies (IgA MAbs) produced from the Peyer's patches and mesenteric lymph nodes of BALB/c mice immunized intragastrically with ricin toxoid. Two IgA MAbs (33G2 and 35H6) were active against ricin's lectin subunit (RTB), and two (23D7 and 25A4) reacted with the toxin's enzymatic subunit (RTA). All four IgA MAbs neutralized ricin in a Vero cell cytotoxicity assay, blocked toxin-induced interleukin-8 release by the human monocyte/macrophage cell line 28SC, and protected polarized epithelial cell monolayers from ricin-mediated protein synthesis inhibition. 33G2 and 35H6 reduced ricin binding to the luminal surfaces of human intestinal epithelial cells to undetectable levels in tissue section overlay assays, whereas 23D7 had no effect on toxin attachment. 23D7 and 25A4 did, however, reduce ricin transcytosis across MDCK II cell monolayers, possibly by interfering with intracellular toxin transport. We conclude that IgA antibodies against RTA and RTB can protect mucosal epithelial cells from ricin intoxication. PMID- 16714578 TI - Differential expression and sequence conservation of the Anaplasma marginale msp2 gene superfamily outer membrane proteins. AB - Bacterial pathogens in the genera Anaplasma and Ehrlichia encode a protein superfamily, pfam01617, which includes the predominant outer membrane proteins (OMPs) of each species, major surface protein 2 (MSP2) and MSP3 of Anaplasma marginale and Anaplasma ovis, Anaplasma phagocytophilum MSP2 (p44), Ehrlichia chaffeensis p28-OMP, Ehrlichia canis p30, and Ehrlichia ruminantium MAP1, and has been shown to be involved in both antigenic variation within the mammalian host and differential expression between the mammalian and arthropod hosts. Recently, complete sequencing of the A. marginale genome has identified an expanded set of genes, designated omp1-14, encoding new members of this superfamily. Transcriptional analysis indicated that, with the exception of the three smallest open reading frames, omp2, omp3, and omp6, these superfamily genes are transcribed in A. marginale-infected erythrocytes, tick midgut and salivary glands, and the IDE8 tick cell line. OMPs 1, 4, 7 to 9, and 11 were confirmed to be expressed as proteins by A. marginale within infected erythrocytes, with expression being either markedly lower (OMPs 1, 4, and 7 to 9) or absent (OMP11) in infected tick cells, which reflected regulation at the transcript level. Although the pfam01617 superfamily includes the antigenically variable MSP2 and MSP3 surface proteins, analysis of the omp1-14 sequences throughout a cycle of acute and persistent infection in the mammalian host and tick transmission reveals a high degree of conservation, an observation supported by sequence comparisons between the St. Maries strain and Florida strain genomes. PMID- 16714580 TI - The major subunit, CfaB, of colonization factor antigen i from enterotoxigenic Escherichia coli is a glycosphingolipid binding protein. AB - Bacterial adherence to mucosal surfaces is an important virulence trait of pathogenic bacteria. Adhesion of enterotoxigenic Escherichia coli (ETEC) to the intestine is mediated by a number of antigenically distinct colonization factors (CFs). One of the most common CFs is CFA/I. This has a fimbrial structure composed of a major repeating subunit, CfaB, and a single tip subunit, CfaE. The potential carbohydrate recognition by CFA/I was investigated by binding CFA/I fimbriated bacteria and purified CFA/I fimbriae to a large number of variant glycosphingolipids separated on thin-layer chromatograms. For both fimbriated bacteria and purified fimbriae, specific interactions could be identified with a number of nonacid glycosphingolipids. These included glucosylceramide, lactosylceramide with phytosphingosine and/or hydroxy fatty acids, neolactotetraosylceramide, gangliotriaosylceramide, gangliotetraosylceramide, the H5 type 2 pentaglycosylceramide, the Lea-5 glycosphingolipid, the Lex-5 glycosphingolipid, and the Ley-6 glycosphingolipid. These glycosphingolipids were also recognized by recombinant E. coli expressing CFA/I in the absence of tip protein CfaE, as well as by purified fimbriae from the same strain. This demonstrates that the glycosphingolipid-binding capacity of CFA/I resides in the major CfaB subunit. PMID- 16714579 TI - Biogenesis of the Actinobacillus actinomycetemcomitans cytolethal distending toxin holotoxin. AB - The cell cycle G2/M specific inhibitor cytolethal distending toxin (CDT) from Actinobacillus actinomycetemcomitans is composed of CdtA, CdtB, and CdtC coded on the cdtA, cdtB, and cdtC genes that are tandem on the chromosomal cdt locus. A. actinomycetemcomitans CdtA has the lipid binding consensus domain, the so-called "lipobox", at the N-terminal signal sequence. Using Escherichia coli carrying plasmid pTK3022, we show that the 16th residue, cysteine, of CdtA bound [3H]palmitate or [)H]glycerol. Further, posttranslational processing of the signal peptide, CdtA, was inhibited using globomycin, an inhibitor of lipoprotein specific signal peptidase II. Fractionation and immunoblotting show the lipid modified CdtA is present in the outer membrane. Immunoprecipitation and the pull down assay of the CDT complex from E. coli carrying a plasmid containing cdtABC demonstrated that the CDT complex in the periplasm is composed of CdtA, CdtB, and CdtC and that the CDT complex in culture supernatant is an N-terminally truncated (36 to 43 amino acids) form of CdtA (CdtA'), CdtB, and CdtC. This suggests that CDT is present as a complex both in the periplasm and the supernatant where CdtA undergoes posttranslation processing to CdtA' in the process of biogenesis and secretion of CDT holotoxin into the culture supernatant. Site-directed mutagenesis of the 16th cysteine residue to glycine in CdtA altered localization of CdtA in the cell and reduced the amount of CDT activity in the culture supernatant. This suggests that CDT forms a complex inside the periplasm for lipid modification where posttranslational processing of CdtA plays an important role for the efficient production of CDT holotoxin into the culture supernatant. PMID- 16714581 TI - Attenuation and persistence of and ability to induce protective immunity to a Staphylococcus aureus aroA mutant in mice. AB - Staphylococcus aureus is the most important etiological agent of bovine mastitis, a disease that causes significant economic losses to the dairy industry. Several vaccines to prevent the disease have been tested, with limited success. The aim of this study was to obtain a suitable attenuated aro mutant of S. aureus by transposon mutagenesis and to demonstrate its efficacy as a live vaccine to induce protective immunity in a murine model of intramammary infection. To do this, we transformed S. aureus RN6390 with plasmid pTV1ts carrying Tn917. After screening of 3,493 erythromycin-resistant colonies, one mutant incapable of growing on plates lacking phenylalanine, tryptophan, and tyrosine was isolated and characterized. Molecular characterization of the mutant showed that the affected gene was aroA and that the insertion occurred 756 bp downstream of the aroA start codon. Complementation of the aroA mutant with a plasmid carrying aroA recovered the wild-type phenotype. The mutant exhibited a 50% lethal dose (1 x 10(6) CFU/mouse) higher than that of the parental strain (4.3 x 10(4) CFU/mouse). The aroA mutant showed decreased ability to persist in the lungs, spleens, and mammary glands of mice. Intramammary immunization with the aroA mutant stimulated both Th1 and Th2 responses in the mammary gland, as ascertained by reverse transcription-PCR, and induced significant protection from challenge with either the parental wild-type or a heterologous strain isolated from a cow with mastitis. PMID- 16714582 TI - Staphylococcus aureus enterotoxin D is secreted in milk and stimulates specific antibody responses in cows in the course of experimental intramammary infection. AB - An enterotoxin D (SED)-producing strain of Staphylococcus aureus was used to infect one mammary gland of each of 17 lactating dairy cows. All glands became infected and shed bacteria over a sampling period of 3 weeks. Serum and milk antibodies specific for SED were monitored by an enzyme-linked immunosorbent assay for 12 weeks. Elevated anti-SED antibodies were detected in all cows after infection, and immunoglobulin of the G2 subclass comprised most of the specific serum response. SED was detected in mastitic milk samples from two cows at levels of 5 to 10 ng/ml. An in vitro lymphocyte proliferation assay showed that SED at levels below 10 pg/ml induced proliferation of bovine lymphocytes and that sheep antiserum specific for SED neutralized this proliferative response. Sera obtained from the cows pre- and postinfection inhibited lymphocyte proliferation at SED concentrations of 10 and 50 ng/ml, respectively. The addition of SED to whole blood or to isolated neutrophils had no significant effect on neutrophil function in vitro. The results show that SED is secreted during mammary gland infection, is mitogenic for bovine lymphocytes, and stimulates the production of specific antibodies. PMID- 16714583 TI - Genomic diversity between strains of the same serotype and multilocus sequence type among pneumococcal clinical isolates. AB - The important human pathogen Streptococcus pneumoniae is known to be a genetically diverse species. We have used comparative genome hybridization (CGH) microarray analysis to investigate this diversity in a collection of clinical isolates including several capsule serotype 14 pneumococci, a dominant serotype among disease isolates. We have identified three new regions of diversity among pneumococcal isolates and, importantly, clearly demonstrate genetic differences between strains of the same multilocus sequence type (ST) and capsule serotype. CGH may therefore, under certain circumstances, prove to be a valuable tool to supplement current typing methods. Finally, we show that these clonal strains with the same serotype and ST behave differently in an animal model. Strains of the same ST and serotype therefore have important genetic and phenotypic differences. PMID- 16714585 TI - Immunological and molecular characterization of susceptibility in relationship to bacterial strain differences in Mycobacterium avium subsp. paratuberculosis infection in the red deer (Cervus elaphus). AB - Johne's disease (JD) infection, caused by Mycobacterium avium subsp. paratuberculosis, represents a major disease problem in farmed ruminants. Although JD has been well characterized in cattle and sheep, little is known of the infection dynamics or immunological response in deer. In this study, typing of M. avium subsp. paratuberculosis isolates from intestinal lymphatic tissues from 74 JD-infected animals showed that clinical isolates of M. avium subsp. paratuberculosis from New Zealand farmed red deer were exclusively of the bovine strain genotype. The susceptibility of deer to M. avium subsp. paratuberculosis was further investigated by experimental oral-route infection studies using defined isolates of virulent bovine and ovine M. avium subsp. paratuberculosis strains. Oral inoculation with high (10(9) CFU/animal) or medium (10(7) CFU/animal) doses of the bovine strain of M. avium subsp. paratuberculosis established 100% infection rates, compared to 69% infection following inoculation with a medium dose of the ovine strain. The high susceptibility of deer to the bovine strain of M. avium subsp. paratuberculosis was confirmed by a 50% infection rate following experimental inoculation with a low dose of bacteria (10(3) CFU/animal). This study is the first to report experimental M. avium subsp. paratuberculosis infection in red deer, and it outlines the strong infectivity of bovine-strain M. avium subsp. paratuberculosis isolates for cervines. PMID- 16714584 TI - CD4+ CD25+ regulatory T cells modulate the T-cell and antibody responses in helicobacter-infected BALB/c mice. AB - Gastric Helicobacter spp. induce chronic gastritis that may lead to ulceration and dysplasia. The host elicits a T helper 1 (Th1) response that is fundamental to the pathogenesis of these bacteria. We analyzed immune responses in Helicobacter-infected, normal mice depleted of CD4+ CD25+ T cells to investigate the in vivo role of regulatory T cells (Tregs) in the modulation of Helicobacter immunopathology. BALB/c and transgenic mice were depleted of CD4+ CD25+ T cells by administration of an anti-CD25 antibody either at the time of infection with Helicobacter or during chronic infection and gastritis. Depletion of CD25+ Tregs prior to and during infection of mice with Helicobacter spp. did not affect either bacterial colonization or severity of gastritis. Depletion of CD25+ Tregs was associated with increased Helicobacter-specific antibody levels and an altered isotype distribution. Paragastric lymph node cells from CD25+ Treg depleted and control infected mice showed similar proliferation to Helicobacter antigens, but only cells from anti-CD25-treated animals secreted Th2 cytokines. CD25+ Tregs do not control the level of gastritis induced by gastric Helicobacter spp. in normal, thymus-intact BALB/c mice. However, CD25+ Tregs influence the cytokine and antibody responses induced by infection. Autoimmune gastritis is not induced in Helicobacter-infected mice depleted of CD25+ Tregs but is induced in CD25+ Treg-depleted mice, which have a higher frequency of autoreactive T cells. PMID- 16714586 TI - MyD88-dependent signaling affects the development of meningococcal sepsis by nonlipooligosaccharide ligands. AB - The Toll-like receptors (TLRs) and the adaptor myeloid differentiation factor 88 (MyD88) are important in the innate immune defenses of the host to microbial infections. Meningococcal ligands signaling via TLRs control inflammatory responses, and stimulation can result in fulminant meningococcal sepsis. In this study, we show that the responses to nonlipooligosaccharide (non-LOS) ligands of meningococci are MyD88 dependent. An isogenic LOS-deficient mutant of the serogroup C meningococcal strain FAM20 caused fatal disease in wild type C57BL/6 mice that was not observed in MyD88-/- mice. Fatality correlated with high proinflammatory cytokine and C5a levels in serum, high neutrophil numbers in blood, and increased bacteremia at 24 h postinfection in the wild-type mice. Infection with the parent strain FAM20 resulted in fatality in 100% of the wild type mice and 50% of the MyD88-/- mice. We conclude that both LOS and another neisserial ligand cause meningococcal sepsis in an in vivo mouse model and confirm that meningococcal LOS can act via both the MyD88- dependent and independent pathways, while the non-LOS meningococcal ligand(s) acts only via the MyD88-dependent pathway. PMID- 16714587 TI - Delineating the requirement for the Borrelia burgdorferi virulence factor OspC in the mammalian host. AB - We previously demonstrated that outer surface protein C (OspC) of Borrelia burgdorferi is essential for establishing mammalian infection. However, the role of OspC in mammalian infection is unknown. Here, we report experiments designed to distinguish between two models of OspC function in the mammalian host: (i) OspC fulfills an essential physiological role for growth and host adaptation or (ii) OspC provides a protective role for evasion of components of the innate immune response. We found that a B. burgdorferi ospC mutant, previously demonstrated to be noninfectious in both immunocompetent and SCID mice, could survive in the relatively immune-privileged environment of dialysis membrane chambers implanted within the peritoneum of a rat. The ospC mutant also adapts to the mammalian environment, as determined by the protein profiles of the chamber cultivated spirochetes. Therefore, OspC does not appear to provide a physiological function for the survival of B. burgdorferi within the mammalian host. The second model, evasion of the innate immune system, was tested by assessing the infectivity of the ospC mutant in mice deficient for myeloid differentiation protein 88 (MyD88). Recent studies have shown that B. burgdorferi is prevented from reaching high cell numbers in the mammalian host by MyD88 dependent signaling pathways. The ospC mutant was incapable of infecting MyD88 deficient mice, suggesting that the role of OspC cannot be related solely to evasion of MyD88-mediated innate immunity. These results reiterate the importance of OspC in mammalian infection and eliminate simple models of function for this enigmatic protein. PMID- 16714588 TI - Borrelia burgdorferi OspC protein required exclusively in a crucial early stage of mammalian infection. AB - This study demonstrates a strict temporal requirement for a virulence determinant of the Lyme disease spirochete Borrelia burgdorferi during a unique point in its natural infection cycle, which alternates between ticks and small mammals. OspC is a major surface protein produced by B. burgdorferi when infected ticks feed but whose synthesis decreases after transmission to a mammalian host. We have previously shown that spirochetes lacking OspC are competent to replicate in and migrate to the salivary glands of the tick vector but do not infect mice. Here we assessed the timing of the requirement for OspC by using an ospC mutant complemented with an unstable copy of the ospC gene and show that B. burgdorferi's requirement for OspC is specific to the mammal and limited to a critical early stage of mammalian infection. By using this unique system, we found that most bacterial reisolates from mice persistently infected with the initially complemented ospC mutant strain no longer carried the wild-type copy of ospC. Such spirochetes were acquired by feeding ticks and migrated to the tick salivary glands during subsequent feeding. Despite normal behavior in ticks, these ospC mutant spirochetes did not infect naive mice. ospC mutant spirochetes from persistently infected mice also failed to infect naive mice by tissue transplantation. We conclude that OspC is indispensable for establishing infection by B. burgdorferi in mammals but is not required at any other point of the mouse-tick infection cycle. PMID- 16714589 TI - Global gene expression profiling of the asymptomatic bacteriuria Escherichia coli strain 83972 in the human urinary tract. AB - Urinary tract infections (UTIs) are an important health problem worldwide, with many million cases each year. Escherichia coli is the most common organism causing UTIs in humans. The asymptomatic bacteriuria E. coli strain 83972 is an excellent colonizer of the human urinary tract, where it causes long-term bladder colonization. The strain has been used for prophylactic purposes in patients prone to more severe and recurrent UTIs. For this study, we used DNA microarrays to monitor the expression profile of strain 83972 in the human urinary tract. Significant differences in expression levels were seen between the in vivo expression profiles of strain 83972 in three patients and the corresponding in vitro expression profiles in lab medium and human urine. The data revealed an in vivo lifestyle of microaerobic growth with respiration of nitrate coupled to degradation of sugar acids and amino acids, with no signs of attachment to host tissues. Interestingly, genes involved in NO protection and metabolism showed significant up-regulation in the patients. This is one of the first studies to address bacterial whole-genome expression in humans and the first study to investigate global gene expression of an E. coli strain in the human urinary tract. PMID- 16714590 TI - Identification of Burkholderia pseudomallei genes required for the intracellular life cycle and in vivo virulence. AB - The bacterial pathogen Burkholderia pseudomallei invades host cells, escapes from endocytic vesicles, multiplies intracellularly, and induces the formation of actin tails and membrane protrusions, leading to direct cell-to-cell spreading. This study was aimed at the identification of B. pseudomallei genes responsible for the different steps of this intracellular life cycle. B. pseudomallei transposon mutants were screened for a reduced ability to form plaques on PtK2 cell monolayers as a result of reduced intercellular spreading. Nine plaque assay mutants with insertions in different open reading frames were selected for further studies. One mutant defective in a hypothetical protein encoded within the Bsa type III secretion system gene cluster was found to be unable to escape from endocytic vesicles after invasion but still multiplied within the vacuoles. Another mutant with a defect in a putative exported protein reached the cytoplasm but exhibited impaired actin tail formation in addition to a severe intracellular growth defect. In four mutants, the transposon had inserted into genes involved in either purine, histidine, or p-aminobenzoate biosynthesis, suggesting that these pathways are essential for intracellular growth. Three mutants with reduced plaque formation were shown to have gene defects in a putative cytidyltransferase, a putative lipoate-protein ligase B, and a hypothetical protein. All nine mutants proved to be significantly attenuated in a murine model of infection, with some mutants being essentially avirulent. In conclusion, we have identified a number of novel major B. pseudomallei virulence genes which are essential for the intracellular life cycle of this pathogen. PMID- 16714592 TI - Members of a Legionella pneumophila family of proteins with ExoU (phospholipase A) active sites are translocated to target cells. AB - Legionella pneumophila replicates within alveolar macrophages, causing a severe pneumonia termed Legionnaires' disease. The bacterium resides within a vacuole that escapes immediate transport to the host lysosome. Instead, the vacuole interacts with the early secretory pathway to establish an environment suitable for rapid multiplication. A type IV secretion system is central to the pathogenicity of the bacterium, and many protein substrates that are translocated by this system to the host cell have been identified. One of these, VipD, was found to interrupt the late secretory pathway when overproduced in Saccharomyces cerevisiae. We independently identified VipD in a previous study and have further characterized this protein as well as its three paralogs. The vipD gene belongs to a family of L. pneumophila open reading frames that are predicted to contain a phospholipase A domain with sequence similarity to the type III-secreted toxin ExoU from Pseudomonas aeruginosa. Similarly to other known translocated proteins of L. pneumophila, VipD is strongly induced in early stationary phase, a time when the bacterium is most virulent. Detergent extraction studies of infected macrophages confirm that VipD is translocated into host cells via the type IV secretion system. A second assay for translocation revealed that two paralogs of VipD, VpdA and VpdB, also have translocation signals recognized by the type IV system. A strain lacking VipD and its three paralogs grew at wild-type rates in murine macrophages, although secondary mutations that cause growth defects in strains lacking VipD accumulate. The quadruple mutant displayed a growth advantage in the amoebal host Dictyostelium discoideum, indicating that the protein family may modulate intracellular growth in a complex fashion. VipD is mildly toxic when overproduced in eukaryotic cells, and the toxicity is partially dependent on the putative phospholipase active site. VipD and its paralogs therefore define a family of translocated proteins that may assist in the establishment of a vacuole suitable for bacterial replication through functioning as a phospholipase. PMID- 16714591 TI - Mycobacterium tuberculosis binding to human surfactant proteins A and D, fibronectin, and small airway epithelial cells under shear conditions. AB - A crucial step in infection is the initial attachment of a pathogen to host cells or tissue. Mycobacterium tuberculosis has evolved multiple strategies for establishing an infection within the host. The pulmonary microenvironment contains a complex milieu of pattern recognition molecules of the innate immune system that play a role in the primary response to inhaled pathogens. Encounters of M. tuberculosis with these recognition molecules likely influence the outcome of the bacillus-host interaction. Here we use a novel fluid shear assay to investigate the binding of M. tuberculosis to innate immune molecules that are produced by pulmonary epithelial cells and are thought to play a role in the lung innate immune response. Virulent and attenuated M. tuberculosis strains bound best to immobilized human fibronectin (FN) and surfactant protein A (SP-A) under this condition. Binding under fluid shear conditions was more consistent and significant compared to binding under static conditions. Soluble FN significantly increased the adherence of both virulent and attenuated M. tuberculosis strains to human primary small airway epithelial cells (SAEC) under fluid shear conditions. In contrast, SP-A and SP-D effects on bacterial adherence to SAEC differed between the two strains. The use of a fluid shear model to simulate physiological conditions within the lung and select for high-affinity binding interactions should prove useful for studies that investigate interactions between M. tuberculosis and host innate immune determinants. PMID- 16714593 TI - Topical application of Escherichia coli-vectored vaccine as a simple method for eliciting protective immunity. AB - We report here that animals can be protected against lethal infection by Clostridium tetani cells and Bacillus anthracis spores following topical application of intact particles of live or gamma-irradiated Escherichia coli vectors overproducing tetanus and anthrax antigens, respectively. Cutaneous gammadeltaT cells were rapidly recruited to the administration site. Live E. coli cells were not found in nonskin tissues after topical application, although fragments of E. coli DNA were disseminated transiently. Evidence suggested that intact E. coli particles in the outer layer of skin may be disrupted by a gammadeltaT-cell-mediated innate defense mechanism, followed by the presentation of E. coli ligand-adjuvanted intravector antigens to the immune system and rapid degradation of E. coli components. The nonreplicating E. coli vector overproducing an exogenous immunogen may foster the development of a new generation of vaccines that can be manufactured rapidly and administered noninvasively in a wide variety of disease settings. PMID- 16714594 TI - Organ-specific role of MyD88 for gene regulation during polymicrobial peritonitis. AB - Sepsis leads to the rapid induction of proinflammatory signaling cascades by activation of the innate immune system through Toll-like receptors (TLR). To characterize the role of TLR signaling through MyD88 for sepsis-induced transcriptional activation, we investigated gene expression during polymicrobial septic peritonitis by microarray analysis. Comparison of gene expression profiles for spleens and livers from septic wild-type and MyD88-deficient mice revealed striking organ-specific differences. Whereas MyD88 deficiency strongly reduced sepsis-induced gene expression in the liver, gene expression in the spleen was largely independent of MyD88, indicating organ-specific transcriptional regulation during polymicrobial sepsis. In addition to genes regulated by MyD88 in an organ-dependent manner, we also identified genes that exhibited an organ independent influence of MyD88 and mostly encoded cytokines and chemokines. Notably, the expression of interferon (IFN)-regulated genes was markedly increased in septic MyD88-deficient mice compared to that in septic wild-type controls. Expression of IFN-regulated genes was dependent on the adapter protein TRIF. These results suggest that the influence of MyD88 on gene expression during sepsis strongly depends on the organ compartment affected by inflammation and that the lack of MyD88 may lead to disbalance of the expression of IFN-regulated genes. PMID- 16714595 TI - Differences in gene expression between the classical and El Tor biotypes of Vibrio cholerae O1. AB - Differences in whole-genome expression patterns between the classical and El Tor biotypes of Vibrio cholerae O1 were determined under conditions that induce virulence gene expression in the classical biotype. A total of 524 genes (13.5% of the genome) were found to be differentially expressed in the two biotypes. The expression of genes encoding proteins required for biofilm formation, chemotaxis, and transport of amino acids, peptides, and iron was higher in the El Tor biotype. These gene expression differences may contribute to the enhanced survival capacity of the El Tor biotype in environmental reservoirs. The expression of genes encoding virulence factors was higher in the classical than in the El Tor biotype. In addition, the vieSAB genes, which were originally identified as regulators of ctxA transcription, were expressed at a fivefold higher level in the classical biotype. We determined the VieA regulon in both biotypes by transcriptome comparison of wild-type and vieA deletion mutant strains. VieA predominantly regulates gene expression in the classical biotype; 401 genes (10.3% of the genome), including those encoding proteins required for virulence, exopolysaccharide biosynthesis, and flagellum production as well as those regulated by sigmaE, are differentially expressed in the classical vieA deletion mutant. In contrast, only five genes were regulated by VieA in the El Tor biotype. A large fraction (20.8%) of the genes that are differentially expressed in the classical versus the El Tor biotype are controlled by VieA in the classical biotype. Thus, VieA is a major regulator of genes in the classical biotype under virulence gene-inducing conditions. PMID- 16714597 TI - Inorganic phosphate induces spore morphogenesis and enterotoxin production in the intestinal pathogen Clostridium perfringens. AB - Clostridium perfringens enterotoxin (CPE) is an important virulence factor for food poisoning and non-food borne gastrointestinal (GI) diseases. Although CPE production is strongly regulated by sporulation, the nature of the signal(s) triggering sporulation remains unknown. Here, we demonstrated that inorganic phosphate (Pi), and not pH, constitutes an environmental signal inducing sporulation and CPE synthesis. In the absence of Pi-supplementation, C. perfringens displayed a spo0A phenotype, i.e., absence of polar septation and DNA partitioning in cells that reached the stationary phase of growth. These results received support from our Northern blot analyses which demonstrated that Pi was able to counteract the inhibitory effect of glucose at the onset of sporulation and induced spo0A expression, indicating that Pi acts as a key signal triggering spore morphogenesis. In addition to being the first study reporting the nature of a physiological signal triggering sporulation in clostridia, these findings have relevance for the development of antisporulation drugs to prevent or treat CPE mediated GI diseases in humans. PMID- 16714596 TI - Infection of human fallopian tube epithelial cells with Neisseria gonorrhoeae protects cells from tumor necrosis factor alpha-induced apoptosis. AB - Following infection with Neisseria gonorrhoeae, bacteria may ascend into the Fallopian tubes (FT) and induce salpingitis, a major cause of infertility. In the FT, interactions between mucosal epithelial cells and gonococci are pivotal events in the pathogen's infection cycle and the inflammatory response. In the current study, primary FT epithelial cells were infected in vitro with different multiplicities of infection (MOI) of Pil+ Opa+ gonococci. Bacteria showed a dose dependent association with cells and induced the secretion of tumor necrosis factor alpha (TNF-alpha). A significant finding was that gonococcal infection (MOI = 1) induced apoptosis in approximately 30% of cells, whereas increasing numbers of bacteria (MOI = 10 to 100) did not induce apoptosis. Apoptosis was observed in only 11% of cells with associated bacteria, whereas >84% of cells with no adherent bacteria were apoptotic. TNF-alpha was a key contributor to apoptosis, since (i) culture supernatants from cells infected with gonococci (MOI = 1) induced apoptosis in naive cultures, suggesting that a soluble factor was responsible; (ii) gonococcal infection-induced apoptosis was inhibited with anti TNF-alpha antibodies; and (iii) the addition of exogenous TNF-alpha induced apoptosis, which was inhibited by the presence of increasing numbers of bacteria (MOI = 10 to 100). These data suggest that TNF-alpha-mediated apoptosis of FT epithelial cells is likely a primary host defense mechanism to prevent pathogen colonization. However, epithelial cell-associated gonococci have evolved a mechanism to protect the cells from undergoing TNF-alpha-mediated apoptosis, and this modulation of the host innate response may contribute to establishment of infection. Understanding the antiapoptotic mechanisms used by Neisseria gonorrhoeae will inform the pathogenesis of salpingitis and could suggest new intervention strategies for prevention and treatment of the disease. PMID- 16714598 TI - Toll-like receptor 2 is required for control of pulmonary infection with Francisella tularensis. AB - Toll-like receptor 2 (TLR2) deficiency enhances murine susceptibility to infection by Francisella tularensis as indicated by accelerated mortality, higher bacterial burden, and greater histopathology. Analysis of pulmonary cytokine levels revealed that TLR2 deficiency results in significantly lower levels of tumor necrosis factor alpha and interleukin-6 but increased amounts of gamma interferon and monocyte chemoattractant protein 1. This pattern of cytokine production may contribute to the exaggerated pathogenesis seen in TLR2-/- mice. Collectively, these findings suggest that TLR2 plays an important role in tempering the host response to pneumonic tularemia. PMID- 16714599 TI - The Babesia bovis merozoite surface antigen 1 hypervariable region induces surface-reactive antibodies that block merozoite invasion. AB - A hypervariable region (HVR) previously identified in the carboxy-terminal one third of the Babesia bovis variable merozoite surface antigen family was more extensively analyzed in merozoite surface antigen 1 (MSA-1) from 16 strains and isolates. The MSA-1 HVR is proline rich and contains three semiconserved motifs nearly identical to those described for the related family member MSA-2. Two MSA 1-specific monoclonal antibodies previously shown to be reactive with the merozoite surface bound to a recombinant construct encoding the HVR, indicating that the HVR is surface exposed and accessible to antibody binding. Importantly, these surface-reactive, HVR-specific monoclonal antibodies were capable of inhibiting merozoite infectivity of the host erythrocyte in vivo. The results indicate that the MSA-1 HVR is involved in erythrocyte invasion and suggest that selection of MSA-1 variants may be driven by invasion-blocking antibodies. PMID- 16714600 TI - Constitutive differences in gene expression profiles parallel genetic patterns of susceptibility to tuberculosis in mice. AB - Interstitial lung macrophages from tuberculosis-susceptible I/St and tuberculosis resistant A/Sn mice demonstrated significant constitutive differences in gene expression levels, whereas in vitro infection of these cells with Mycobacterium tuberculosis had only a modulatory impact on gene expression. We conclude that intrinsic gene expression profiles are an important determinant of tuberculosis pathogenesis in mice. PMID- 16714601 TI - Localization of the C3-Like ADP-ribosyltransferase from Staphylococcus aureus during bacterial invasion of mammalian cells. AB - The C3stau2 exoenzyme from Staphylococcus aureus is a C3-like ADP ribosyltransferase which possesses no specific receptor-binding domain or translocation unit required for entry in target cells where its substrate is located. Here we show that C3stau2 can reach its target after invasion of staphylococci in eukaryotic cells without needing translocation. PMID- 16714602 TI - Infectivity of the highly transformable BBE02- lp56- mutant of Borrelia burgdorferi, the Lyme disease spirochete, via ticks. AB - Infectious Borrelia burgdorferi strains that have increased transformability with the shuttle vector pBSV2 were recently constructed by inactivating the gene encoding BBE02, a putative restriction-modification gene product expressed by the linear plasmid lp25 (Kawabata et al., Infect. Immun. 72:7147-7154, 2004). The absence of the linear plasmid lp56, which carries another putative restriction modification gene, further enhanced transformation rates. The infectivity of these mutants was assessed previously in mice that were inoculated with needle and syringe and was found to be equivalent to that of wild-type spirochetes. Here we examined the infectivity of spirochetes to ticks after capillary inoculation of Ixodes scapularis nymphs and the subsequent spirochetal infectivity to mice via ticks by using B. burgdorferi B31 clonal isolates lacking lp56 and/or BBE02. The absence of lp56 (but not BBE02) correlated with a lower number of spirochetes in ticks after feeding on mice; this plasmid thus may play a role, albeit not an essential one, in supporting spirochetal survival in the feeding tick. Importantly, however, the absence of lp56 and BBE02 did not detectably influence infectivity to mice via ticks. PMID- 16714603 TI - Isolation of a human monoclonal antibody with strong neutralizing activity against diphtheria toxin. AB - We isolated a human monoclonal antibody against diphtheria toxin (DT). It bound to fragment B with a binding activity (Kd) of 3.01 nM. The neutralizing activity assayed by the rabbit skin test was estimated to be 73,600 IU/g. This could be used as a therapeutic drug against DT in place of the traditional equine sera. PMID- 16714604 TI - Development of luminescent Mycobacterium avium subsp. paratuberculosis for rapid screening of vaccine candidates in mice. AB - Mycobacterium avium subsp. paratuberculosis is a slowly growing mycobacterial species, requiring 6 to 8 weeks of culture before colonies can be counted visually. Here, we describe the development of luminescent M. avium subsp. paratuberculosis expressing luxAB genes of Vibrio harveyi and its use for vaccine testing in an experimental mouse model, replacing fastidious CFU counting by rapid luminometry. PMID- 16714606 TI - Cardiac imaging 2006. PMID- 16714605 TI - The ABC transporter protein OppA provides protection against experimental Yersinia pestis infection. AB - The identification of Yersinia pestis as a potential bioterrorism agent and the emergence of antibiotic-resistant strains have highlighted the need for improved vaccines and treatments for plague. The aim of this study was to evaluate the potential for ATP-binding cassette (ABC) transporter proteins to be exploited as novel vaccines against plague. Western blotting of ABC transporter proteins using sera from rabbits immunized with killed whole Y. pestis cells or human convalescent-phase sera identified four immunologically reactive proteins: OppA, PstS, YrbD, and PiuA. Mice immunized with these proteins developed antibody to the immunogen. When the immunized mice were challenged with Y. pestis, the OppA immunized mice showed an increased time to death compared to other groups, and protection appeared to correlate with the level of immunoglobulin G antibody to OppA. PMID- 16714607 TI - Beta-blockers for cardiac CT: a primer for the radiologist. AB - OBJECTIVE: The objective of this article is to describe a protocol for the administration of beta-blockers for cardiac CT. A low and regular heart rate is necessary for optimal visualization of the coronary arteries on CT and can be achieved by the administration of medications. CONCLUSION: Beta-blockers can be safely given, orally or IV, to most patients to lower the heart rate for cardiac CT. A protocol can be implemented and patients can be screened for certain contraindications to allow successful administration of these medications by radiologists. PMID- 16714608 TI - MDCT angiography of acute chest pain: evaluation of ECG-gated and nongated techniques. AB - OBJECTIVE: The objective of our study was to compare MDCT angiography protocols used in patients with acute chest pain caused by vascular, nonvascular, and cardiac abnormalities. SUBJECTS AND METHODS: In four groups of 20 patients with chest pain each, four MDCT protocols were used based on monitoring vascular attenuation: pulmonary embolism (150 H at pulmonary artery), aortic dissection (200 H at aortic arch), chest pain (200 H at pulmonary artery), and chest pain with ECG gating (150 H at pulmonary artery). Vascular enhancement was assessed by attenuation measurements taken from locations in the pulmonary artery (n = 3) and thoracic aorta (n = 4). The appearance of the coronary artery in regard to opacification and motion was assessed on a scale of 1 to 5 (best). RESULTS: The mean pulmonary artery and aorta attenuation (372 H and 352 H, respectively) was significantly higher (p < 0.005, Student's t test) and the number of vessel attenuation points measuring less than 200 H (1/140) was significantly smaller (p < 0.001, chi-square test) in the chest pain compared with the dissection (318 H, 310 H; 16/140), gated chest pain (304 H, 286 H; 17/14), and pulmonary embolism (302 H, 220 H; 28/140) groups. The median coronary artery visualization score was 4; the proximal regions received a significantly (p < 0.005, Mann-Whitney test) higher grade compared with the middle and distal regions (medians, 5, 4, and 2, respectively). Artifacts were noted on the gated scans. CONCLUSION: The chest pain protocol can be used to assess both the pulmonary arteries and the thoracic aorta, whereas the ECG-gating protocol appears to be a promising adjunct for a comprehensive single chest pain protocol. PMID- 16714609 TI - Cardiovascular function in multi-ethnic study of atherosclerosis: normal values by age, sex, and ethnicity. AB - OBJECTIVE: MRI provides accurate and high-resolution measurements of cardiac anatomy and function. The purpose of this study was to describe the imaging protocol and normal values of left ventricular (LV) function and mass in the Multi-Ethnic Study of Atherosclerosis (MESA). SUBJECTS AND METHODS: Eight hundred participants (400 men, 400 women) in four age strata (45-54, 55-64, 65-74, 75-84 years) were chosen at random. Participants with the following known cardiovascular risk factors were excluded: current smoker, systolic blood pressure > 140 mm Hg, diastolic blood pressure > 90 mm Hg, fasting glucose > 110 mg/dL, total cholesterol > 240 mg/dL, and high-density lipoprotein (HDL) cholesterol < 40 mg/dL. Cardiac MR images were analyzed using MASS software (version 4.2). Mean values, SDs, and correlation coefficients in relationship to patient age were calculated. RESULTS: There were significant differences in LV volumes and mass between men and women. LV volumes were inversely associated with age (p < 0.05) for both sexes except for the LV end-systolic volume index. For men, LV mass was inversely associated with age (slope = -0.72 g/year, p = 0.0021), but LV mass index was not associated with age (slope = -0.179 g/m2/year, p = 0.075). For women, LV mass (slope = -0.15 g/year, p = 0.30) and LV mass index (slope = 0.0044 g/m2/year, p = 0.95) were not associated with age. LV mass was the largest in the African-American group (men, 181.6 +/- 35.8 [SD] g; women, 128.8 +/- 28.1 g) and was smallest in the Asian-American group (men, 129.1 +/- 20.0 g; women, 89.4 +/- 13.3 g). CONCLUSION: The normal LV differs in volume and mass between sexes and among certain ethnic groups. When indexed by body surface area, LV mass was independent of age for both sexes. Studies that assess cardiovascular risk factors in relationship to cardiac function and structure need to account for these normal variations in the population. PMID- 16714610 TI - MDCT assessment of right ventricular systolic function. AB - OBJECTIVE: The objective of our study was to validate using MDCT for the assessment of right ventricular (RV) function. MDCT with retrospective ECG gating was performed in 15 patients being evaluated for suspected cardiovascular disease. Echocardiography was performed for comparison. The MDCT images were reconstructed at 20 phase points over the cardiac cycle. The end-diastolic and end-systolic volumes of both ventricles were measured. Stroke volumes and ejection fractions were calculated from these data. CONCLUSION: RV volumes can be accurately assessed using MDCT. PMID- 16714611 TI - MDCT determination of volume and function of the left ventricle: are short-axis image reformations necessary? AB - OBJECTIVE: Determination of left ventricular (LV) volumes and global function parameters from MDCT data sets is usually based on short-axis reformations from primarily reconstructed axial images, which prolong postprocessing time. The aim of this study was to evaluate the feasibility of LV volumetry and global LV function assessment from axial images in comparison with short-axis image reformations. SUBJECTS AND METHODS: This study consisted of 20 patients with either coronary artery disease or dilated cardiomyopathy. We evaluated MDCT results using cine MRI as the reference technique. RESULTS: LV end-diastolic volume (LVEDV) and end-systolic volume (LVESV) were significantly overestimated by the axial MDCT approach in comparison with volume measurements from short-axis CT image reformations. The mean LV ejection fraction (LVEF) was not significantly different (41.2% vs 42.7%). Short-axis and axial MDCT determination of LVEF revealed a systematic underestimation by a mean +/- SD of -2.1% +/- 3.6% versus 3.6% +/- 8.2%, respectively, when compared with LVEF values based on cine MRI. The interobserver variability for volume and function measurements from axial images (LVEDV = 8.5%, LVESV = 10.8%, LVEF = 9.6%) was slightly higher than those measurements from short-axis reformations (LVEDV = 7.2%, LVESV = 9.5%, LVEF = 8.7%). The mean total evaluation time was significantly shorter using axial images (14.1 +/- 3.9 min) compared with short-axis reformations (16.9 +/- 5.2 min) (p < 0.05). CONCLUSION: Determination of LV volumes and assessment of global LV function from axial MDCT image reformations is feasible and time efficient. This approach might be a clinically useful alternative to established short-axis based measurements in patients with normal or near-normal LV function. A progressive underestimation of LVEF with increasing LV volumes may limit the clinical applicability of the axial approach in patients with dilated cardiomyopathy. PMID- 16714612 TI - Synergy of MDCT and cine MRI for the evaluation of cardiac motility. AB - OBJECTIVE: The objective of our study was to validate the feasibility of the synergistic use of cardiac MR and CT data sets for ventricular motility analysis and to correlate measurement variability with underlying heart rate. SUBJECTS AND METHODS: Twenty patients underwent concurrent ECG-gated MDCT and MRI for evaluation of ventricular motility, expressed as ventricular wall thickening and motion. Initially, individual measurement repetition series were analyzed by determining intraobserver variability and detecting intraobserver bias related to heart rates. Subsequently, absolute measurement differences of CT or MR data were statistically evaluated. Finally, absolute measurement differences were correlated with underlying heart rates by curve estimation regression. RESULTS: Analysis of measurement reproducibility proved that data variability was dependent on only the anatomic localization of the analyzed ventricular segment, not on the imaging technique used or underlying heart rate, in normofrequent patients. Comparing MR and CT image data sets, no statistically significant differences were identified when ventricular motility was evaluated based on data sets of either imaging technique in normofrequent patients. Tachycardic frequencies, above 100 beats per minute, led to exponential error propagation due to insufficient temporal resolution of the current CT technology. CONCLUSION: This study proved that cardiac motility assessment based on ECG-gated CT and MR data sets resulted in comparable ventricular function results for normofrequent patients; however, the high spatial resolution of cardiac MDCT cannot compensate for the lack of temporal resolution in patients with tachycardia, thus emphasizing the necessity of reporting ventricular motility analysis results in combination with heart rate to allow consideration of this possible cause for measurement variation. PMID- 16714613 TI - MDCT of the coronary arteries: feasibility of low-dose CT with ECG-pulsed tube current modulation to reduce radiation dose. AB - OBJECTIVE: The objective of our study was to show the feasibility of coronary CT using low kilovoltage (80 kV) combined with ECG-pulsed tube current modulation in selected patients. CONCLUSION: This study showed the combined effect of lowering the kilovoltage setting (80 kV) and using an automatic modulation technique (ECG pulsed tube current modulation) for coronary CT. Radiation dose exposure can be reduced by up to 88% for slim patients without impairing image quality. PMID- 16714614 TI - Detection of myocardial bridging with ECG-gated MDCT and multiplanar reconstruction. AB - OBJECTIVE: The aim of this study was to evaluate the incidence of myocardial bridging in 626 patients examined with MDCT angiography of the coronary arteries. MATERIALS AND METHODS: Six hundred twenty-six patients who were referred to Florence Nightingale and Ataturk University Hospitals were involved in this study. These patients had atypical chest pain, symptoms suggestive of coronary artery disease, or no significant cardiac complaint. Patients were in sinus rhythm and were premedicated with metoprolol tartrate (5 mg/mL IV bolus) to decrease the heart rate and nitroglycerin (5 mg sublingual 1 min before the examination) to dilate the coronary arteries. MDCT was performed on two different 16-MDCT scanners. RESULTS: Among the 626 patients, 22 cases (3.5%) of myocardial bridging were detected. Fifteen cases of myocardial bridging (2.4%) were located at the middle third of the left anterior descending coronary artery (LAD), five (0.8%) were at the distal third of the LAD, and two (0.3%) were at the proximal third of the LAD. In these patients, the length of tunneled artery was between 6 and 22 mm, with a mean of 17 mm, and the depth of tunneled artery was between 1.2 and 3.3 mm, with a mean of 2.5 mm. CONCLUSION: We found the incidence of myocardial bridging in this patient group to be 3.5%. This result is in agreement with some of the angiographic studies in the literature. Our study showed that MDCT is a reliable and noninvasive tool for diagnosing coronary myocardial bridging. After evaluating resource axial images, it is necessary to also evaluate the sagittal multiplanar reconstruction images for myocardial bridging. PMID- 16714615 TI - Early postoperative assessment of coronary artery bypass graft patency and anatomy: value of contrast-enhanced 16-MDCT with retrospectively ECG-gated reconstructions. AB - OBJECTIVE: The objective of our study was to assess early postoperative patency and anatomy of off-pump coronary artery bypass grafts (CABGs) using retrospectively ECG-gated MDCT. CONCLUSION: Retrospectively ECG-gated MDCT is a promising noninvasive technique with which to assess early postoperative patency and anatomy of CABGs. PMID- 16714616 TI - MRI of coronary vessel walls using radial k-space sampling and steady-state free precession imaging. AB - OBJECTIVE: The objective of our study was to investigate the impact of radial k space sampling and steady-state free precession (SSFP) imaging on image quality in MRI of coronary vessel walls. SUBJECTS AND METHODS: Eleven subjects were examined on a 1.5-T MR system using three high-resolution navigator-gated and cardiac-triggered 3D black blood sequences (cartesian gradient-echo [GRE], radial GRE, and radial SSFP) with identical spatial resolution (0.9 x 0.9 x 2.4 mm3). The signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), vessel wall sharpness, and motion artifacts were analyzed. RESULTS: The mean SNR and CNR of the coronary vessel wall were improved using radial imaging and were best using radial k-space sampling combined with SSFP imaging. Vessel border definition was similar for all three sequences. Radial k-space sampling was found to be less sensitive to motion. Consistently good image quality was seen with the radial GRE sequence. CONCLUSION: Radial k-space sampling in MRI of coronary vessel walls resulted in fewer motion artifacts and improved SNR and CNR. The use of SSFP imaging, however, did not result in improved coronary vessel wall visualization. PMID- 16714617 TI - Computer-aided detection and evaluation of lipid-rich plaque on noncontrast cardiac CT. AB - OBJECTIVE: Noncontrast electron beam CT (EBCT) and MDCT are established for the assessment of calcified plaque, but not lipid-rich plaque. We developed software to identify lipid-rich plaque with noncontrast electron beam tomography (EBT) and MDCT. MATERIALS AND METHODS: A computer algorithm was developed to automatically find contiguous lipid-rich lesions with voxel intensities below a calculated patient-specific lipid threshold. Lipid density and lipid inhomogeneity in Hounsfield units were calculated in the proximal left coronaries of three populations: 34 low-risk patients (low-risk group < 6% Framingham risk score, no calcium), 31 high-risk patients (high-risk group > 20% Framingham risk score, no calcium), and 37 patients with calcified plaque (calcium group). RESULTS: The mean lipid density was -19.6 +/- 3.0 (SD) H in the low-risk group, -25.3 +/- 8.2 H in the high-risk group, and -34.3 +/- 13.0 H in the calcium group (p < 0.05). The mean lipid inhomogeneity was 17.7 +/- 3.6 H in the low-risk group, 21.5 +/- 5.5 H in the high-risk group, and 29.0 +/- 7.6 H in the calcium group (p < 0.05). The mean interscan variability in lipid density and lipid inhomogeneity were 2.0 +/- 3.3 H and 2.1 +/- 3.6 H, respectively. In five patients, the locations of lipid-rich plaque correlated well with available intravascular sonography findings. CONCLUSION: Our method may be able to identify lipid-rich plaque on noncontrast cardiac CT. PMID- 16714618 TI - CT analysis of the growth rate of aortic diameter affected by acute type B intramural hematoma. AB - OBJECTIVE: The purpose of this study was to evaluate the growth rate of aortic diameter affected by acute type B intramural hematoma and the factors that influence its enlargement. MATERIALS AND METHODS: Fifty-four patients were entered into this study, and regular follow-up CT studies (mean +/- SD, 46.9 +/- 27.2 months; range, 5-136 months) were performed. The affected aortas and iliac arteries were divided into five segments. A total of 127 segments were evaluated (aortic arch, n = 47; descending thoracic aorta, n = 51; suprarenal abdominal aorta, n = 24; infrarenal abdominal aorta, n = 3; and iliac artery, n = 2). The growth rate of each segment was obtained on CT. The factors influencing increase in the diameter and growth rate--age, sex, diabetes mellitus, atherosclerotic disease, history of smoking 20 years, chronic renal failure, blood pressure, initial diameter of the lumen, the presence of blood flow in the false lumen- were evaluated by univariate analysis and a multivariate logistic regression model. RESULTS: Twenty (37.0%) of 54 patients had one or more segments that increased in size during the follow-up period. Of a total of 127 segments, 35 (27.6%) increased in size, and for all, the mean growth rate was -0.5 +/- 2.9 mm/year. This negative growth rate represents shrinkage. The mean growth rates within the first year and after 1 year from onset were -2.2 +/- 5.7 and 0.4 +/- 3.2 mm/year, respectively, and a significant difference was observed between them (p < 0.0001). An initial diameter of 40 mm or greater and the presence of blood flow in the false lumen were significant risk factors for an increase in the diameter, as confirmed by univariate and multivariate analyses. CONCLUSION: In patients with type B intramural hematoma, the affected aortas did not show a high incidence of enlargement during the follow-up period, but the affected aortas tended to increase in size after 1 year from onset. An initial diameter of 40 mm or greater and the presence of blood flow in the false lumen were important risk factors for enlargement during the follow-up period. PMID- 16714619 TI - Acute and chronic pulmonary emboli: angiography-CT correlation. AB - OBJECTIVE: The objective of our study was to review the classic direct and indirect angiographic signs of acute and chronic pulmonary embolism (PE) and correlate these findings with MDCT. CONCLUSION: CT and angiography have complementary roles in the accurate diagnosis of acute and chronic thromboembolic disease. Conventional angiography should be used as a problem-solving technique after CT angiography has been performed because CT angiography is less invasive. PMID- 16714621 TI - Radiological reasoning: incidentally discovered liver mass. AB - OBJECTIVE: A 49-year-old woman presented to the emergency department after a fall in which she sustained a right subcapital hip fracture. During her hospital stay she developed abdominal pain, and a hypoechoic liver mass was found on sonography. Multiphase CT showed a hepatic mass with brisk arterial phase enhancement, rapid washout on the portal venous phase, and delayed phase hypodensity. The final pathology diagnosis was hepatocellular carcinoma. CONCLUSION: Incidental lesions are frequently discovered during routine radiographic evaluations. Correlation with clinical history and additional confirmatory imaging is essential for appropriate diagnosis and management. PMID- 16714623 TI - AJR Teaching File: chronic nausea and vomiting with abdominal pain. PMID- 16714624 TI - AJR Teaching File: enlarging splenic mass after nephrectomy. PMID- 16714625 TI - AJR Teaching File: acute abdominal pain after combined kidney and pancreas transplantation. PMID- 16714627 TI - Defecography for the diagnosis of abnormalities in patients with fecal incontinence. PMID- 16714629 TI - Treatment of sacral insufficiency fractures. PMID- 16714631 TI - Wet versus dry laser printers for copying digital mammograms. PMID- 16714632 TI - Appropriate use of CT scanning in acute appendicitis. PMID- 16714633 TI - Female breast radiation exposure. PMID- 16714634 TI - The legacy of "Visualization of the chambers of the heart, the pulmonary circulation, and the great blood vessels in man". PMID- 16714635 TI - Patient-controlled room air insufflation versus automated carbon dioxide delivery for CT colonography. AB - OBJECTIVE: Our objective was to prospectively compare colonic distention and patient comfort at screening CT colonography (CTC) with patient-controlled room air (RA) insufflation versus automated carbon dioxide (CO2) delivery. SUBJECTS AND METHODS: Two hundred eight adults undergoing CTC were randomized to RA or CO2 (104 per method). Colonic distention was prospectively assessed by segment using a 4-point scale (1 = optimal, 2 = adequate, 3 = inadequate, 4 = collapsed). Adequacy of combined supine/prone segmental evaluation was also recorded. Patients provided discomfort ratings on a 0-10 scale (0-1 = none/insignificant, 2 3 = minimal, 4-6 = intermediate, 7-10 = significant) before, during, and after the procedure. RESULTS: No significant differences in segmental distention were observed in the prone position between techniques. In the supine position, CO2 resulted in significantly greater distention in the sigmoid, descending, and transverse segments (p < 0.01). After combined supine/prone assessment, two/104 (1.9%) and three/104 (2.9%) patients were judged to have an inadequately evaluated segment on RA and CO2, respectively (four sigmoid, one transverse). Mean discomfort scores for RA and CO2 were 3.97 and 5.08 during the examination (p < 0.01); 0.91 and 0.42 immediately after (p < 0.01); 0.51 and 0.25 15 min later (p < 0.05); and 0.15 and 0.04 2 hours later (p < 0.01), respectively. During active distention, 19 (18.3%) and 33 (31.7%) patients reported significant transient discomfort with RA and CO2, respectively (p < 0.05). Beyond 15 min, only two (1.9%) patients with RA and no patients with CO2 had a discomfort level higher than 3. CONCLUSION: Although patient-controlled RA insufflation and automated CO2 delivery each resulted in negligible postprocedure discomfort and reliable colonic distention, CO2 was better for both categories. PMID- 16714636 TI - CT features of hepatic venoocclusive disease and hepatic graft-versus-host disease in patients after hematopoietic stem cell transplantation. AB - OBJECTIVE: We conducted this study to evaluate whether CT scans could be used to differentiate hepatic venoocclusive disease from hepatic graft-versus-host disease in patients treated with hematopoietic stem cell transplantation. SUBJECTS AND METHODS: We retrospectively evaluated 18 patients (eight women, 10 men; mean age, 42.4 years) after hematopoietic stem cell transplantation with biopsy-proven hepatic venoocclusive disease (n = 5), hepatic graft-versus-host disease (n = 6), or both (n = 7). Two radiologists reviewed abdominal and pelvic CT scans for hepatomegaly (> 18 cm), splenomegaly (> 13 cm), size of main portal and right hepatic veins, presence of periportal edema, gallbladder wall edema, hydropic gallbladder, ascites, and small-bowel wall thickening. CT and histopathology findings were correlated using analysis of variance and Fisher Free-man-Holton tests. RESULTS: Ascites and periportal edema were present in all five patients with venoocclusive disease, but of six patients with graft-versus host disease, ascites was seen in two (p < 0.05) and periportal edema in only one (p < 0.05). Small-bowel wall thickening was encountered in five patients with graft-versus-host disease and in none with venoocclusive disease (p < 0.05). The right hepatic vein diameter in patients with venoocclusive disease (mean, 0.27 cm) was significantly smaller than the right hepatic vein diameter in patients with graft-versus-host disease (mean, 0.87 cm; p < 0.05). CONCLUSION: In patients treated with hematopoietic stem cell transplantation, CT findings of periportal edema, ascites, and a narrow right hepatic vein suggest venoocclusive disease rather than graft-versus-host disease. Small-bowel wall thickening suggests graft versus-host disease. PMID- 16714637 TI - Comparison of CT during arterial portography and MR during arterial portography in the detection of liver metastases. AB - OBJECTIVE: This study compared MR during arterial portography (MRAP) with CT during arterial portography (CTAP) with regard to the detection and differentiation of liver metastases before surgery. MATERIALS AND METHODS: Fifteen patients with liver metastases were enrolled before surgery according to the guidelines of our institutional review board and good clinical practice. After mesentericography, unenhanced scans (Volume Zoom) were performed initially. For CTAP, the contrast medium was injected through the superior mesenteric artery. Images were acquired in portal and delayed enhancement. The MR protocol (1.5 T; Magnetom Symphony) started with T1-weighted fast low-angle shot (FLASH) T2-weighted turbo spin echo (TSE). MRAP followed with gadolinium-enhanced dynamic T1-weighted 3D FLASH. Delayed-phase T1-weighted 2D FLASH axial images were performed 2 min after IV injection of the contrast medium. Qualitative and quantitative evaluation of CTAP and MRAP was performed by three blinded radiologists regarding the number of lesions and their size, localization, and differential diagnosis. RESULTS: The overall sensitivity in detecting liver metastases was 97% with MRAP and 93% with CTAP (p > 0.05, not significant [n.s.]). The specificity was calculated to be 97% for MRAP and 82% for CTAP (p < 0.0001, statistically significant [s.s.]). The differences in sensitivity were more accentuated if only lesions 10 mm or smaller were considered (95% vs 88%, p > 0.05, n.s.), for which the respective specificities were 95% and 80% (p < 0.0014, s.s.). Improvements in sensitivity and specificity were associated with a higher lesion-to-liver contrast-to-noise ratio (59.4 +/- 51.0 for MRAP vs 10.4 +/ 7.3 for CTAP) and resulted in higher diagnostic confidence in the differential diagnosis of liver lesions (p < 0.001, s.s.) and better interobserver agreement (median kappa value, 0.88 vs 0.63). CONCLUSION: MRAP proved to be a reliable method in the preoperative detection of small liver metastases in particular, with a higher sensitivity and specificity than CTAP. If organizational difficulties of MRAP can be overcome, MRAP could be considered instead of CTAP in the preoperative invasive evaluation of metastatic liver disease. PMID- 16714638 TI - Evaluation of primary malignancies of the liver using contrast-enhanced sonography: correlation with pathology. AB - OBJECTIVE: Our purpose was to investigate the correlation of contrast-enhanced sonographic patterns with the histopathology of primary malignancies of the liver. MATERIAL AND METHODS: Of the 318 contrast-enhanced sonographic examinations performed between April and September 2004, 95 patients with 117 lesions confirmed by surgery or biopsy pathology were reviewed in this study. We analyzed the enhancement time and patterns according to the types and degrees of pathologic differentiation. RESULTS: All 65 moderately to poorly differentiated hepatocellular carcinomas (HCCs) enhanced in the arterial phase, and 96.9% (63 lesions) of them quickly washed out in the portal venous phase. All 32 well differentiated HCCs enhanced in the arterial phase, and 50.0% (16 lesions) washed out slowly during the late phase. The washout time of the two differentiated types was significantly different (p < 0.05). Seventy-five percent of the clear cell carcinomas (12/16) enhanced in the arterial phase, 25% (4/16) did not enhance until the portal venous phase, and 31.3% (5/16) of the clear cell carcinomas washed out slowly during the late phase. The enhancement and washout times of clear cell carcinomas were significantly different than those of moderately to poorly differentiated HCCs (p < 0.05). All four intrahepatic cholangiocarcinomas presented the same patterns as moderately to poorly differentiated HCCs: enhanced in the arterial phase and quickly washed out in the portal venous phase. Among the 72 lesions enhanced homogeneously, 46 lesions (63.9%) were smaller than 3 cm. Thirty-seven lesions (82.2%) of the 45 lesions that enhanced heterogeneously were larger than 3 cm (including the four cholangiocarcinomas). Larger lesions were mainly less differentiated. There were significant differences in tumor sizes in regard to cellular differentiations and types. CONCLUSION: Our study showed that the enhancement manifestations of primary malignancies of the liver are related to pathologic types and grades. Contrast-enhanced sonograms may provide the histopathologic information for malignant tumors of the liver. PMID- 16714639 TI - The celiac ganglia: anatomic study using MRI in cadavers. AB - OBJECTIVE: Our objective was to facilitate the in vivo identification of the celiac ganglia on MRI by using MRI to identify the celiac ganglia in cadavers. CONCLUSION: MRI can show the celiac ganglia accurately in cadavers when the ganglia are large and labeled with gadolinium. The findings in cadavers can be a reference for identifying the celiac ganglia in vivo. PMID- 16714640 TI - Abdominal MRI at 3.0 T: the basics revisited. AB - OBJECTIVE: The purpose of our article is to describe the underlying physics concepts of abdominal MRI at 3.0 T and their impact on signal-to-noise ratio, susceptibility artifacts, chemical shift artifacts, and dielectric effects. CONCLUSION: Abdominal MR sequence protocols optimized for 1.5-T scanners should not be transferred to 3.0 T without substantial modification. In addition, specific patient groups--for example, large patients with ascites--are not well suited to undergo an abdominal MRI study at 3.0 T. PMID- 16714641 TI - MRI of focal splenic lesions without and with dynamic gadolinium enhancement. AB - OBJECTIVE: Our purpose was to identify the MR features of focal splenic lesions with an emphasis on enhancement patterns. The addition of a contrast-enhanced dynamic sequence to unenhanced MR images improves the study of focal splenic lesions. The analysis of the MR features along with the clinical history permits either the characterization of the most common splenic lesions or improvement in the detection of malignant diseases. CONCLUSION: Dynamic contrast-enhanced MRI improves the detection and characterization of focal splenic lesions. PMID- 16714642 TI - Abdominal metastases of medulloblastoma related to a ventriculoperitoneal shunt. PMID- 16714643 TI - Prospective multicenter trial evaluating a novel method of characterizing focal liver lesions using contrast-enhanced sonography. AB - OBJECTIVE: The purpose of this study was to assess the clinical value and potential impact of SonoVue-enhanced sonography in the characterization of focal liver lesions. SUBJECTS AND METHODS: This study included 127 patients with 82 malignant and 52 benign lesions in the liver. Contrast-enhanced sonography was performed using nonlinear imaging modes at low mechanical index (0.1-0.3) to enable real-time visualization of arterial, portal, and late-phase enhancement. Digital recordings of unenhanced sonography and contrast-enhanced sonography were reviewed by on-site investigators and two off-site blinded interpreters. The final diagnosis was based on consensus interpreting of all examinations by another two expert observers with access to CT, MRI, and histologic data; the diagnostic accuracy of contrast-enhanced sonography in identifying the lesion as benign, malignant, or indeterminate and as actual tumor type was compared with baseline sonography. RESULTS: For on-site investigators, contrast-enhanced sonography reduced the number of indeterminate diagnoses by 67% and improved the sensitivity and specificity to 90.2% and 80.8%, respectively (p < 0.001). For off site interpreters, contrast-enhanced sonography reduced the number of indeterminate diagnoses by 51-56% (p < 0.001); significantly improved sensitivity and specificity to 90.8-95.4% and 83.7-89.8%, respectively (p < 0.001); eliminated observers' variability (kappa coefficient: 0.66-0.77); and showed no significant difference in all comparisons in the analysis of lesions measuring less than 1.5 cm, 1.5-2.5 cm, and all sizes combined. Contrast-enhanced sonography did not rely on availability of clinical history to enable the diagnoses, and it reduced the need for further imaging investigations 23.7% to 90.4%. CONCLUSION: Contrast-enhanced sonography improves the characterization of focal liver lesions and may limit the need for further investigations. PMID- 16714644 TI - Comparison of visual and quantitative analysis for characterization of insonated liver tumors after microbubble contrast injection. AB - OBJECTIVE: The objective of our study was to compare diagnostic performance of visual and quantitative analysis for the characterization of liver tumors insonated at low transmit power after microbubble contrast agent injection. SUBJECTS AND METHODS: This series comprised 166 liver tumors (1-5 cm in diameter) in 166 patients (99 men, 67 women; mean age +/- SD, 58 +/- 11 years) scanned at low transmit power (mechanical index: 0.1-0.14) after sulfur hexafluoride-filled microbubble injection. Digital cine clips recorded at the arterial phase (10-40 sec after contrast injection) and late phase (100-300 sec) were analyzed to characterize liver tumors as benign or malignant. Visual analysis was performed by three independent blinded reviewers who evaluated enhancement patterns at the arterial phase and subjective tumor conspicuity at the late phase. Quantitative analysis of videotape intensity (VI: gray-scale levels, 0-255) was performed to calculate objective tumor conspicuity at the late phase: (VI(tumor) - VI(liver)) / VI(liver). RESULTS: Characteristic enhancement patterns were observed in malignant tumors (peripheral rimlike) and benign tumors (peripheral nodular or central and spoke-wheel-shaped). Malignant (n = 95) versus benign (n = 71) tumors differed for subjective (median value: -1 vs 1, respectively) and objective conspicuity at the late phase (-0.6 vs 0.15, respectively; p = 0.001, Mann Whitney U test) due to persistent microbubble uptake in benign tumors. Diagnostic performance of visual (odds ratio: reviewer 1 = 4.28, reviewer 2 = 10.18, reviewer 3 = 9.56) and quantitative (odds ratio: 89.33) analyses differed significantly in the characterization of liver tumors (p = 0.01, chi-square test). CONCLUSION: Quantitative analysis revealed higher diagnostic performance than visual analysis to characterize liver tumors insonated at low transmit power after microbubble contrast agent injection. PMID- 16714645 TI - Quadruple-phase MDCT of the liver in patients with suspected hepatocellular carcinoma: effect of contrast material flow rate. AB - OBJECTIVE: The purposes of this study were to evaluate the effect of contrast material flow rate (3 mL/sec vs 5 mL/sec) on the detection and visualization of hepatocellular carcinoma (HCC) with MDCT and the safety profile of iodixanol at different injection rates. SUBJECTS AND METHODS: In a prospective, randomized multicenter trial, 97 patients (83 men and 14 women, with a mean age of 64 years) suspected of having HCC underwent quadruple-phase (double arterial, portal venous, delayed phase) 4-16-MDCT. Patients were randomized to receive iodixanol, 320 mg I/mL (1.5 mL/kg body weight), at a flow rate of 3 mL/sec (48 patients) or 5 mL/sec (49 patients). Qualitative (lesion detection, image quality) and quantitative (liver and aortic enhancement, tumor-liver contrast) analyses and safety assessment were performed. RESULTS: Overall, 145 HCCs were detected in the 5 mL/sec group and 100 HCCs in the 3 mL/sec group (p < 0.05). More lesions equal to or less than 1 cm were detected at 5 mL/sec (33 vs 16 lesions). The late arterial phase showed significantly more lesions than the early, arterial phase (133 vs 100 and 96 vs 67 lesions, respectively, p < 0.0001). Hyperattenuating HCCs were better visualized in the late arterial phase at 5 mL/sec (excellent visualization: 54% vs 27%). Using a flow of 5 mL/sec did not increase the rate of patient discomfort or contrast media-related adverse events. Most discomfort in both groups was of mild intensity and there was no severe discomfort. CONCLUSION: For detection of HCC with MDCT, a higher flow rate of 5 mL/sec is recommended. Visualization of hyperattenuating HCC is improved with no greater discomfort or adverse events. PMID- 16714646 TI - Preoperative evaluation of resectability of Klatskin tumor with 16-MDCT angiography and cholangiography. AB - OBJECTIVE: The objective of our study was to evaluate prospectively the preoperative use of 16-MDCT angiography and cholangiography in determining the resectability of Klatskin tumors. CONCLUSION: Preoperative MDCT angiography and cholangiography gave a good assessment of the degree of vascular and biliary involvement of the Klatskin tumor. PMID- 16714647 TI - Focal nodular hyperplasia: lesion evaluation using 16-MDCT and 3D CT angiography. AB - OBJECTIVE: The purpose of this article is to present the imaging features of focal nodular hyperplasia (FNH) using MDCT and 3D CT angiography. CONCLUSION: MDCT with advanced image processing is a powerful tool that may be utilized to identify the imaging features of FNH. These include the presence of large feeding arteries and draining veins, pseudocapsule, central scar, and septations. These features can help in the differentiation of this benign lesion from other hypervascular lesions without the need for additional imaging, biopsy, or surgery. PMID- 16714648 TI - Polyp measurement using CT colonography: agreement with colonoscopy and effect of viewing conditions on interobserver and intraobserver agreement. AB - OBJECTIVE: This article presents inter- and intraobserver agreement for estimates of polyp diameter using CT colonography, including the effects of different visualization displays and prior experience. MATERIALS AND METHODS: Four observers, three of whom had prior experience with CT colonography, estimated the maximum diameter of 48 polyps using three different visualization displays: 2D colonography window, 2D abdominal window, and 3D surface rendering. Each re measured a subset of 10 polyps. Polyps measured 2 to 12 mm according to a colonoscopic reference. Inter- and intraobserver agreement and agreement with the reference measurement were determined using the Bland-Altman method, paired Student's t testing, analysis of variance, and analysis of covariance (ANCOVA), and by calculating the components of variance. RESULTS: CT measurements overestimated polyp diameter, a phenomenon found least using the 2D abdominal display. Generally, 95% limits of agreement encompassed different size categories for individual polyps: the widest spanned 14.6 mm (-4.6 mm to 10.0 mm) for an experienced observer using the 3D display. When using the 2D abdominal display, no significant difference was found between estimates and the reference value for the other two experienced observers (p = 0.83 and 0.23). All the observers' measurements were significantly different from the reference when using the 3D display (p < 0.001). The novice was significantly different from the experienced observers in some analyses. Inter- and intraobserver agreement were poorest for the 3D display. CONCLUSION: Measurement of polyp diameter from CT colonography is subject to variation contingent on the observer's experience and the viewing display used. Although 3D visualization display is commonly used for polyp detection, it should not be used for measurement. PMID- 16714649 TI - Polyp volume versus linear size measurements at CT colonography: implications for noninvasive surveillance of unresected colorectal lesions. AB - OBJECTIVE: Proposed surveillance of unresected medium-sized (6.0-9.9 mm) polyps will require reliable detection of small incremental changes. The purpose of this study was to assess the reproducibility of linear versus volume measurements of polyp size at CT colonography (CTC) and to correlate results with a hemispheric model. MATERIALS AND METHODS: The study group consisted of 30 polyps on supine and prone CTC data sets. Measurements were performed separately by two experienced radiologists using the same CTC system (Vitrea 2), resulting in 120 linear and volume measurements. Linear size was defined as the longest dimension among the 2D multiplanar reconstruction views. Semiautomated volume determination required manual tracing of polyp boundaries on 2D multiplanar reconstruction views. The relative error between reviewers 1 and 2 was defined as 100 x (/D1 - D2/)/D(ave) for linear size (D) and as 100 x (/V1 - V2)/V(ave) for volume (V), where ave is average. RESULTS: The mean relative error for linear size and volume measurement was 10.4% +/- 10.7% and 16.9% +/- 13.2%, respectively. Median linear size and volume of polyps were 9.4 mm and 270 mm3, respectively. CTC-derived volumes for medium-sized polyps closely approximated hemispheric volume (V = (pi/12) x D(b), where b =3.13, r2 = 0.90). Small incremental changes in hemispheric size result in a 3:1 relative change in volume versus diameter, such that a 1-mm diameter change in a medium-sized hemispheric polyp results in a relative change in linear size and volume ranging from approximately 11-18% and 31-53%, respectively. CONCLUSION: Because changes in polyp volume are amplified compared with linear dimension, volume measurement rather than diameter measurement will better allow detection of small incremental changes in polyp size using CTC. PMID- 16714650 TI - Flat polyps of the colon: detection with 16-MDCT colonography--preliminary results. AB - OBJECTIVE: This study evaluates the ability of contrast-enhanced, 16-MDCT colonography to display flat colonic lesions when a very narrow slice thickness (1 mm) is used. CONCLUSION: Less than 50% of flat lesions in our population could be visualized by blinded and unblinded review. Before they could be visualized, flat lesions were 2 mm or greater in height and 7 mm or greater in diameter. Lesions with a height of 1 mm or less were not seen on CT colonography. Contrast enhancement, location on a haustral fold, and abnormal 2D and 3D morphology contributed to lesion conspicuity. PMID- 16714651 TI - Bloodborne metastatic tumors to the gastrointestinal tract: CT findings with clinicopathologic correlation. AB - OBJECTIVE: In this essay, we illustrate the CT features of bloodborne metastases to the gastrointestinal tract from various malignancies with an emphasis on clinicopathologic correlation. CONCLUSION: Familiarity with CT findings of bloodborne metastases to the gastrointestinal tract and an understanding of the disease spread pattern in common primary cancers will be helpful not only in detecting metastatic disease but also in minimizing the possibility of mistaking metastasis for another metachronous malignancy. PMID- 16714652 TI - Esophagopleural fistula secondary to esophageal wall ballooning and thinning after pneumonectomy: findings on chest CT and esophagography. PMID- 16714653 TI - Utility of 16-MDCT angiography for comprehensive preoperative vascular evaluation of laparoscopic renal donors. AB - OBJECTIVE: Our objective was to determine the efficacy of 16-MDCT angiography in preoperative evaluation of vascular anatomy of laparoscopic renal donors. METHODS AND MATERIALS: Fifty-five consecutive renal donors (25 men and 30 women) underwent 16-MDCT angiography followed by donor nephrectomy. In the arterial and nephrographic phases, images were acquired with 60% overlap and 0.6-mm reconstruction in both phases after 120 mL of iohexol was injected at 4 mL/sec. On a 3D workstation, images were evaluated retrospectively by two abdominal imagers blinded to surgical results with respect to number and branching pattern of renal arteries and major and minor renal veins. These CT angiography results were compared with surgical findings. RESULTS: The surgically confirmed sensitivity of both reviewers (1 and 2) using the MDCT data for detection of renal arteries was 98.5% (65 of 66), and accuracies were 97.0% for reviewer 1 and 95.5% for reviewer 2. Sensitivity and accuracy detection of renal veins was 97% (61 of 63) and 98% (62 of 63) for reviewer 1 and reviewer 2, respectively. Sensitivity and accuracy detection of early arterial bifurcation (< 2 cm from aorta) was 100% (14 of 14), and sensitivity in detection of late venous confluence (< 1.5 cm from aorta) was 100% (8 of 8). All major renal venous variants were identified; reviewer 1 identified 78% (18 of 23) minor venous variants, and reviewer 2 identified 83% (19 of 23) minor venous variants. There were no hemorrhagic complications at surgery. Excellent agreement between reviewers (kappa = 0.92-0.97) was achieved for detection of normal and variant anatomy. CONCLUSION: 16-MDCT angiography enabled excellent preoperative detection of arterial anatomy and venous laparoscopic donor nephrectomy. PMID- 16714654 TI - Comparison of 16-MDCT and MRI for characterization of kidney lesions. AB - OBJECTIVE: The objective of our study was to compare the diagnostic performance of 16-MDCT with that of MRI in the characterization of kidney lesions. SUBJECTS AND METHODS: Twenty-eight patients with kidney lesions detected with sonography and requiring further evaluation were examined. MDCT was performed in the unenhanced, arterial, and portal venous phases. MRI was performed at 1.5 T with T2- and T1-weighted and dynamic gadolinium-enhanced sequences. Consensus reading was done by two radiologists. Image quality was rated on a four-point scale. Classification of lesions as surgical or nonsurgical was done with five levels of confidence, and it was required that a definite diagnosis be assigned to each lesion. The 1997 TNM classification was used for staging. Statistical analysis was done by receiver operating characteristic analysis or paired Student's t test. Histologic or follow-up findings at least 12 months after the primary diagnosis served as the standard of reference. RESULTS: The image quality of MDCT (mean grade, 2.79 on a 0-3 scale) was superior to that of MRI (1.93; p < 0.01). The area under the curve for differentiating surgical from nonsurgical lesions was 0.979 for MDCT and 0.957 for MRI with resulting sensitivity and specificity values of 92.3% and 96.3% for MDCT and 92.3% and 91.3% for MRI. Sensitivity and specificity for definite classification of the lesions were 93.8% and 68.4% for MDCT and 93.8% and 71.4% for MRI. CONCLUSION: Both MDCT and MRI are excellent for differentiating surgical from nonsurgical kidney lesions. Both methods have low specificity for the differentiation of benign from malignant lesions. PMID- 16714655 TI - IV contrast administration for CT: a survey of practices for the screening and prevention of contrast nephropathy. AB - OBJECTIVE: No widely accepted practice guidelines exist for the screening and prevention of contrast nephropathy in patients receiving iodinated IV contrast agents for CT examinations. Issues include screening to identify at-risk patients, discriminatory levels of renal insufficiency beyond which IV contrast material is contraindicated, and the use of preventive measures. We surveyed radiologists to identify the current practice patterns and to determine those areas in which guidelines are most needed. MATERIALS AND METHODS: Surveys were e mailed to 2,000 radiologists. The survey included questions on the issues just described and respondent demographics. RESULTS: The response rate was 21%. Serum creatinine is the most commonly used screening method: 92% of respondents for inpatient examinations and 66% for outpatient examinations. Only 2% use estimated creatinine clearance. The average threshold serum creatinine value used to determine that patients should not receive IV contrast material is 1.5 mg/dL in 35%, 1.7 mg/dL in 27%, and 2.0 mg/dL in 31% (mean, 1.78 mg/dL). Diabetes slightly lowers the threshold creatinine (mean, 1.68 mg/dL). Fewer than 30% of respondents frequently administer IV contrast material to patients with a renal transplant or multiple myeloma. The most commonly used preventive measures include hydration (93%), reduction of contrast dose (77%), and administration of acetylcysteine (39%). Overall, no important differences were found among practice settings or level of specialization. CONCLUSION: A wide variation of practice patterns is apparent in the screening and prevention of contrast nephropathy. In some cases, these patterns conflict with recommendations from the literature. The results of this study identify opportunities for further research and areas in need of improved practice guidelines. PMID- 16714656 TI - Clinical value of MDCT in the diagnosis of coronary artery disease in patients with a low pretest likelihood of significant disease. AB - OBJECTIVE: The aim of this study was to evaluate the clinical value of MDCT in the diagnosis of coronary artery disease in a population having a low pretest likelihood of significant disease. SUBJECTS AND METHODS: Sixty-four patients with suspected coronary artery disease and a low pretest likelihood of significant disease according to the criteria of the American Heart Association underwent both MDCT of the heart and quantitative conventional coronary angiography (QCA). MDCT examinations were performed on a 16-MDCT scanner. CT data sets were evaluated on a per-patient basis and a per-segment basis and were classified as indicating no disease, nonsignificant disease (stenoses 50%). Sensitivity, specificity, negative predictive value (NPV), and positive predictive value (PPV) of 16-MDCT in the detection or exclusion of significant and nonsignificant coronary artery disease were evaluated on both per-patient and per-segment bases. RESULTS: Regarding the success rate of 16-MDCT, 94% (60/64) of patients and 92% (388/420) of vessel segments were of sufficient quality for diagnosis. In the remaining 60 patients evaluated, QCA revealed significant coronary artery disease, nonsignificant disease, and no disease in 8.3% (5/60), 75.0% (45/60), and 16.7% (10/60) of cases, respectively, on a per-patient basis, and in 1.3% (5/388), 23.2% (90/388), and 75.5% (293/388) of cases, respectively, on a per-segment basis. The sensitivity, specificity, NPV, and PPV of 16-MDCT for the detection of significant coronary artery disease were 80.0%, 94.5%, 98.1%, and 57.1%, respectively, on a per-patient basis, and 80.0%, 99.2%, 99.7%, and 57.1% on a per segment basis. CONCLUSION: In a population having a low pretest likelihood of significant coronary artery disease, 16-MDCT shows a moderate to high sensitivity and high NPV for the detection or exclusion of significant disease, but has a somewhat reduced PPV compared with QCA. PMID- 16714657 TI - SENSE or k-MAG to accelerate free breathing navigator-guided coronary MR angiography. AB - OBJECTIVE: The purpose of this study was to assess the relative merits of reducing the scanning time of navigator-guided (NAV) coronary MR angiography by including, both independently and in combination, two time-saving strategies: k space weighted motion-adapted gating (k-MAG) and sensitivity encoding (SENSE, factor = 2). SUBJECTS AND METHODS: Coronary arteries of 21 healthy subjects were imaged with four NAV MR angiography sequences: conventional NAV sequence, NAV with the addition of SENSE, NAV with the addition of k-MAG, and NAV with a combination of SENSE and k-MAG. All imaging parameters including the magnetization preparation schemes, prescribed spatial resolution, and acquisition duration per R-R interval were identical for all techniques. The total scanning time, navigator efficiency, visible length of the coronary artery, and subjective image quality were used as metrics for evaluating the performance of the techniques. RESULTS: The results show that the addition of k-MAG to NAV coronary MR angiography (with or without SENSE) improved scan efficiency and decreased scanning time by an average of 17% without compromising the length of coronary artery visible or the image quality. The addition of SENSE to the NAV technique (with or without k-MAG) reduces the scanning time by an average of 50%. CONCLUSION: While the average image quality of coronary arteries was unaffected by the addition of k-MAG to navigator techniques, there was a slight reduction in image quality scores for the navigator sequence with SENSE. Identification of the proximal coronary arteries was not hampered by the addition of k-MAG, SENSE, or both to the NAV coronary MR angiography sequence. PMID- 16714658 TI - Diagnostic performance of MDCT for detecting aortic valve regurgitation. AB - OBJECTIVE: This study evaluates whether ECG-gated 16-MDCT coronary angiography provides a reliable imaging technique for detecting aortic regurgitation (AR). SUBJECTS AND METHODS: We examined 71 patients prospectively with 16-MDCT angiography using retrospective ECG gating during the mid-to-end diastolic phase. A visible central valvular leakage area was considered as a diagnostic criterion for AR. The central valvular leakage area was quantified in patients with positive transthoracic echocardiography (TTE). We compared 16-MDCT to Doppler TTE, the accepted diagnostic standard based on semiquantitative regurgitation jet analysis (grade 0-3+ for mild, moderate, and severe). RESULTS: The overall sensitivity of 16-MDCT for the identification of patients with AR was 81%. The specificity was 91%, the negative predictive value was 70%, and the positive predictive value was 95%. Of the 71 patients, 48 had AR determined by TTE, and 16 MDCT correctly detected AR in 39 of those 48 patients. The sensitivity of 16-MDCT for the detection of patients with moderate and severe AR (grade > or = 1.5+) was 95%, and the specificity was 96% (20 of 21 patients). The sensitivity of 16-MDCT for identification of patients with mild AR (grade < or = 1+) was 70%, and the specificity was 92% (19 of 27 patients). Quantification of the central valvular leakage area was not possible in 50% of cases with AR by TTE because of valve calcifications. CONCLUSION: Sixteen-MDCT coronary angiography provides an accurate, noninvasive imaging technique to detect moderate and severe aortic regurgitation (grade > or = 1.5+). However, severe valve calcifications and mild AR limit its results. PMID- 16714659 TI - MR findings in cardiac amyloidosis. AB - OBJECTIVE: The objective of our study was to describe a combination of features on MRI specific to cardiac amyloidosis. CONCLUSION: Cardiac amyloidosis is a common cause of infiltrative heart disease. The combination of subtle widespread heterogeneous myocardial enhancement on delayed postcontrast inversion recovery T1-weighted images, which may initially be dismissed as a technical error, with ancillary features of restrictive cardiac disease is highly suggestive of cardiac amyloidosis. PMID- 16714660 TI - Experience in 207 combined MRI examinations for acute pulmonary embolism and deep vein thrombosis. AB - OBJECTIVE: The purpose of this study was to prospectively assess the feasibility and quality of combined MRI examinations consisting of thoracic MRI for suspected pulmonary embolism (PE) and MR venography for deep vein thrombosis (DVT), to assess the diagnostic yield of a combined examination for detecting thromboembolism compared with each component alone, and to retrospectively assess the concordance of duplex sonography and MR venography. SUBJECTS AND METHODS: Two hundred twenty-one consecutive patients (119 men, 102 women; mean age, 51 years; range, 31-86 years) with suspected PE were examined using a multitechnique thoracic MRI protocol (real-time MRI using true fast imaging with steady-state precession [FISP], perfusion MRI, and MR angiography) followed by stepping-table MR venography. RESULTS: Two hundred twenty-one thoracic MRI examinations were performed. Two hundred eighteen MR venography examinations were scheduled, of which five (2.3%) were not performed for clinical or technical reasons and six were not performed after negative thoracic MRI. Among 207 combined examinations, PE was diagnosed in 76 and DVT in 78 examinations. Thirteen patients without PE showed DVT; thus, MR venography detected 17% additional cases of thromboembolism. Agreement with duplex sonography was good at the upper leg (kappa = 0.87-0.89) but moderate at the pelvis (kappa = 0.59-0.65). CONCLUSION: A combined "one-stop shopping" MRI approach for PE and DVT was routinely feasible and detected 17% more cases of thromboembolism compared with separate examinations. MRI may be considered a second-line technique to avoid contraindications to CT but also a primary comprehensive technique for diagnosing thromboembolism. PMID- 16714661 TI - Pulmonary lesions associated with visceral larva migrans due to Ascaris suum or Toxocara canis: imaging of six cases. AB - OBJECTIVE: The objective of our study was to evaluate chest radiographic and CT findings of patients with pulmonary lesions associated with visceral larva migrans due to Ascaris suum or Toxocara canis. CT investigation was focused on the location, size, contour, and internal features of the lesions; migration of lesions; mediastinal lymphadenopathy; and pleural effusion. CONCLUSION: Pulmonary visceral larva migrans appears on CT as multifocal subpleural nodules with halo or ground-glass opacities and ill-defined margins. PMID- 16714662 TI - Percutaneous cryotherapy of the thorax: safety considerations for complex cases. PMID- 16714664 TI - Focusing in on dose reduction: the FDA perspective. PMID- 16714663 TI - Thin-Section CT 12 Months After the Diagnosis of Severe Acute Respiratory Syndrome in Pediatric Patients. AB - OBJECTIVE: The objective of our study was to report the thin-section CT findings 12 months after the diagnosis of severe acute respiratory syndrome (SARS) in pediatric patients who had recovered clinically but had persistent abnormal CT findings 6 months after the diagnosis. The clinical data for these patients were correlated to identify risk factors that might increase the likelihood of the development of CT abnormalities. SUBJECTS AND METHODS: The study involved an extended 12-month thin-section CT follow-up of 16 of 47 pediatrics patients with SARS coronavirus-associated pneumonia proven serologically (21 girls and 26 boys; age range, 1.5-17 years; median age, 13.6 years). Patients' clinical information, the extent of radiographic opacification during the acute phase of illness, and conventional pulmonary function test results on follow-up were obtained for correlation. The clinical parameters were compared with other pediatric SARS patients who had normal CT findings at the 6-month follow-up. RESULTS: Fifteen patients still had abnormal CT findings 12 months after diagnosis, all of whom were older than 10 years (age range, 10-17 years). In seven patients with previous residual ground-glass opacification at the 6-month follow-up, two showed persistent changes and three had a reticular pattern in the area of the previously detected abnormality, whereas two showed complete resolution. The extent of air trapping remained similar to that at the 6-month follow-up in nine of 11 patients while two showed a slight increase in the same segments. Parenchymal scars remained unchanged from the 6- to 12-month follow-up in all six patients with that finding. None of our patients showed any evidence of bronchiectasis or bronchial wall thickening. Lymphopenia (p = 0.03), extent of radiographic opacification at acute illness (p = 0.047), and duration of use of ribavirin (p = 0.03) were significant risk factors in predicting whether abnormal CT features persisted 12 months after diagnosis. CONCLUSION: We found that 32% of the children (15/47) affected with SARS showed thin-section CT abnormalities up to 12 months after diagnosis despite clinical remission and unremarkable pulmonary function assessment. Persistent CT abnormalities are more likely to develop in patients who are older and who present with more severe disease. The CT changes in children with SARS are, however, minor. PMID- 16714665 TI - Radiation dose to the female breast from 16-MDCT body protocols. AB - OBJECTIVE: The objective of our study was to determine the radiation dose to the female breast from current 16-MDCT body examinations. MATERIALS AND METHODS: Metal oxide semiconductor field effect transistor (MOSFET) detectors were placed in four quadrants of the breast of a female-configured anthropomorphic phantom to determine radiation dose to the breast. Imaging was performed on a 16-MDCT scanner (LightSpeed, GE Healthcare) using current clinical protocols designed to assess pulmonary embolus (PE) (140 kVp, 380 mA, 0.8-sec rotation, 16 x 1.25 mm collimation), appendicitis (140 kVp, 340 mA, 0.5-sec rotation, 16 x 0.625 mm collimation), and renal calculus (140 kVp, 160 mA, 0.5-sec rotation, 16 x 0.625 mm collimation). RESULTS: Radiation dose to the breast ranged from 4 to 6 cGy for the PE protocol and up to 1-2 cGy in the inferior aspect of the right breast and lateral aspect of the left breast for the appendicitis protocol. The renal calculus protocol yielded less than 150 microGy absorbed breast dose. CONCLUSION: Current clinical chest and abdomen protocols result in vairable radiation doses to the breast. The magnitude of exposure may have implications for imaging strategies. PMID- 16714666 TI - Contrast-enhanced breast MRI in patients with suspicious microcalcifications on mammography: results of a multicenter trial. AB - OBJECTIVE: The objective of our study was to test dynamic MRI in evaluating mammographically detected suspicious microcalcifications. MATERIALS AND METHODS: One hundred twelve patients with mammographically detected microcalcifications with BI-RADS category 5 (n = 78) or 4 (n = 34) lesions were studied at 17 centers a using 3D gradient-echo dynamic coronal technique (< or = 3 mm thickness) and 0.1 mmol/kg of gadoteridol. A pathologic sample was obtained in all cases. Agreement between the major diameter measured on mammography, MRI, or both and the major diameter measured at pathologic examination was calculated in 62 cases. RESULTS: Of the 112 lesions, pathologic examination revealed 37 benign lesions, 33 ductal carcinoma in situ (DCIS), and 42 invasive carcinomas. The specificity of MRI for benign lesions was 68%. Considering the subgroups of calcifications alone and calcifications associated with masses, the specificity values became 79% and 33%, respectively. The sensitivity of MRI for DCIS was 79%. Analysis of the two subgroups showed sensitivity values of 68% for calcifications alone and of 1% for calcifications associated with masses. The sensitivity for invasive carcinomas was 93%. Analysis of the two subgroups showed sensitivity values to be 92% for calcifications alone and 94% for calcifications associated with masses. Considering the overall results, the sensitivity of MRI was 87%; specificity, 68%; positive predictive value, 84%; negative predictive value, 71%; and accuracy, 80%. Considering the subgroups of calcifications alone and calcifications associated with masses, the sensitivity values became 80% and 97%; the positive predictive values, 86% and 82%; the negative predictive values, 71% and 75% (95% confidence interval [CI], 0.19-0.99); and the accuracy values, 80% and 82% (95% CI, 0.66-0.92), respectively. An odds ratio (OR) of 13.54 (95% CI, 5.20-35.28) showed a raised risk of malignant breast tumor in subjects with positive MR examination of mammographically detected suspicious clusters of microcalcifications. The statistical analysis on each subgroup showed an OR of 15.07 (95% CI, 4.73-48.08) for calcifications alone and an OR of 14.00 (95% CI, 1.23-158.84) for calcifications associated with masses. Any significant improvement in the predictive ability of dynamic MRI depending on the extent of calcifications on mammography was not proved. Considering the 62 cases of proved malignancy with measured maximal diameter at pathologic examination, both mammography and MR examination seem to overestimate tumor extent. CONCLUSION: The not-perfect sensitivity of MRI (87%), when applying our interpretation criteria and imaging sequences, is a crucial point that prevents us from clinical use of MRI in the diagnosis of mammographically detected microcalcifications. PMID- 16714667 TI - MRI-guided vacuum-assisted breast biopsy with a handheld portable biopsy system. AB - OBJECTIVE: The purpose of this study was to evaluate a compact portable 10-gauge handheld battery-operated vacuum-assisted biopsy system for MRI-guided breast biopsy. CONCLUSION: The compact portable battery-operated biopsy system can be used successfully for MRI-guided core breast biopsy and is an alternative to current systems. PMID- 16714668 TI - Optimization of CT angiography of the carotid artery with a 16-MDCT scanner: craniocaudal scan direction reduces contrast material-related perivenous artifacts. AB - OBJECTIVE: The objective of our study was to compare the effect of a caudocranial scan direction versus a craniocaudal scan direction on arterial enhancement and perivenous artifacts in 16-MDCT angiography of the supraaortic arteries. SUBJECTS AND METHODS: Eighty consecutive patients (51 men; mean age, 62 years; age range, 28-89 years) underwent scanning in the caudocranial direction (group 1; n = 40) or the craniocaudal direction (group 2; n = 40). All patients received 80 mL of contrast material followed by a 40-mL saline chaser bolus, both administered IV at 4 mL/sec. Bolus tracking was used. Attenuation inside the arterial lumen was measured at intervals of 1 sec throughout the data set. Attenuation in the superior vena cava (SVC) was measured. Contrast material-related perivenous artifacts were graded on a scale of 0-3 (none to extensive). RESULTS: Attenuation in the ascending aorta, carotid bifurcation, and intracranial arteries was slightly lower in group 2 versus group 1 (231 +/- 64 H, 348 +/- 52 H, and 258 +/- 48 H vs 282 +/- 43 H, 381 +/- 73 H, and 291 +/- 77 H, respectively; p < 0.05). Maximum and mean arterial attenuations were slightly lower in group 2 versus group 1 (369 +/- 58 H and 303 +/- 48 H vs 401 +/- 71 H and 334 +/- 58 H; p < 0.05). Attenuation in the SVC was much lower in group 2 versus group 1 (169 +/- 39 H vs 783 +/- 330 H; p < 0.001). Mean streak artifact score was much lower in group 2 versus group 1 (1.3 +/- 0.9 vs 2.5 +/- 0.6; p < 0.001). CONCLUSION: Use of a craniocaudal scan direction results in slightly lower attenuation of the carotid artery and much lower attenuation of the SVC. Streak artifacts are significantly reduced. This technique allows better evaluation of the ascending aorta and supraaortic arteries. PMID- 16714669 TI - Hybrid peripheral 3D contrast-enhanced MR angiography of calf and foot vasculature. AB - OBJECTIVE: The objective of our study was to describe hybrid peripheral (HyPer) 3D contrast-enhanced MR angiography (CE-MRA) using sagittal acquisition with parallel imaging of the calf and foot station. The benefit of a dedicated sagittal 3D CE-MRA acquisition of the calf and foot was evaluated by assessing the degree of venous contamination and its diagnostic quality compared with standard bolus chase 3D CE-MRA alone. MATERIALS AND METHODS: Fifty-three patients (99 legs) were scanned with a 1.5-T MR system equipped with a dedicated bilateral lower extremity phased-array coil. First, high-resolution 3D CE-MRA images of the calves and feet were obtained using two separate sagittal slabs with parallel imaging, with a resulting voxel size of 1.4 x 1.0 x 1.0 mm3. Second, standard bolus chase 3D CE-MRA was performed from the abdomen and pelvis station to the calf-foot station. Images were interpreted by two radiologists. The calf-foot arterial trees were divided into 12 segments. Each segment was characterized as diagnostic or nondiagnostic. The degree of venous contamination was assessed as interfering with the diagnosis or not. Paired Student's t test and Wilcoxon's signed rank test were used to test for statistically significant differences between the techniques. RESULTS: For the left leg (n = 48), the mean number (+/- SD) of diagnosed arterial segments for HyPer 3D CE-MRA was 9.2 +/- 2.3 and for bolus chase 3D CE-MRA, 7.1 +/- 4.2 (p < or = 0.0004). For the right leg (n = 51), the corresponding values were 9.4 +/- 2.2 and 7.6 +/- 3.5 (p < or = 0.0005), respectively. For bolus chase 3D CE-MRA, venous contamination interfered with the diagnosis in 24 of 99 legs, whereas with HyPer 3D CE-MRA, there was no interference. Selective analysis of the dorsalis pedis arteries showed that the number of diagnostic vessels was 62 (62.6%) of 99 for HyPer 3D CE-MRA and 13 (13.1%) of 99 for bolus chase 3D CE-MRA. CONCLUSION: HyPer 3D CE-MRA is an alternative method for time-resolved high-resolution peripheral CE-MRA in evaluating the trifurcation and feet vessels with no venous contamination. PMID- 16714670 TI - MDCT versus digital radiography in the evaluation of bone healing in orthopedic patients. AB - OBJECTIVE: Assessment of bone healing in orthopedic patients is usually monitored by radiographs in two views. The purpose of our study was to compare multiplanar reconstructions from MDCT data sets with digital radiographs for assessing the extent of bone healing. MATERIALS AND METHODS: Forty-three orthopedic patients (19 women, 24 men) who underwent MDCT and radiography after arthrodesis, fractures, or spinal fusions were included in our study. MDCT was performed on an MX 8000IDT scanner and served as the gold standard. The technical parameters were adapted to the anatomic region. A bone algorithm for reconstruction was used (3,500/600 H). Multiplanar reconstructions were calculated in two orthogonal planes. All patients underwent digital radiography on a Multix FD system in two views according to standard procedures. Multiplanar reconstructions and radiographs were analyzed by two musculoskeletal radiologists in a consensus interpretation to determine bone healing using a semiquantitative approach. RESULTS: In 27 patients (63%), MDCT and digital radiography were concordant with regard to the extent of bone healing, whereas in 16 patients (37%) the results were not concordant. In eight patients (19%) digital radiographs underestimated the extent of bone healing, whereas in another eight patients (19%) they overestimated the degree of fusion. CONCLUSION: MDCT using high-quality 2D reformatting is recommended as the primary imaging technique for the evaluation of bone healing. PMID- 16714671 TI - Relationship between bone marrow edema and development of symptoms in patients with osteonecrosis of the femoral head. AB - OBJECTIVE: The purpose of this study was to determine the significance of risk factors on MR images for predicting the outcome of patients with osteonecrosis of the femoral head. SUBJECTS AND METHODS: Eighty-three asymptomatic or minimally symptomatic hips in 61 consecutive patients were followed up prospectively. Inclusion criteria included osteonecrosis of the femoral head identified by typical MRI findings, with no radiographic evidence of progression of collapse, and Harris hip score of 85 points or more. Every 3 months, the patients underwent clinical and radiographic examination, and MR images were obtained at 6- to 12 month intervals. Follow-up continued until worsening of hip pain with Harris hip score of less than 70 points or surgery. Asymptomatic hips were followed up for at least 24 months. The mean clinical and radiographic follow-up period after the initial diagnosis was 60 months (range, 3-168 months). RESULTS: Thirty-six (43%) of the 83 hips were symptomatic at the last follow-up. Bone marrow edema was present in 28 hips (34%) during the follow-up period. Twenty-seven (96%) of the 28 hips were symptomatic, and bone marrow edema significantly correlated with worsening of hip pain (p < 0.0001). The necrotic volume of hips with bone marrow edema was significantly larger than those without bone marrow edema (p < 0.0001). Bone marrow edema was found to be the most significant risk factor for worsening of pain (p < 0.0001). CONCLUSION: Bone marrow edema strongly correlated with necrotic volume and was the most significant risk factor for worsening of hip pain. A large necrotic volume of 30% or more may be the second useful indicator for predicting future worsening of hip pain. PMID- 16714672 TI - MRI comparison of periprosthetic structures around zirconium knee prostheses and cobalt chrome prostheses. AB - OBJECTIVE: The purpose of our study was to compare reviewer confidence and interobserver agreement in the evaluation of MR images of periprosthetic structures around zirconium total knee prostheses and cobalt chrome prostheses. MATERIALS AND METHODS: Three board-certified radiologists blinded to prosthesis type used identical MRI protocols to independently evaluate 21 total knee prostheses: 14 zirconium prostheses and seven cobalt chrome prostheses. The radiologists evaluated the following eight parameters: integrity of the medial and lateral collateral ligaments, the quadriceps and the patellar tendons, presence of a joint effusion, and for evidence of periprosthetic osseous signal abnormality around each of the three components. The reviewers gave their degree of confidence in evaluating each of the findings on a five-point scale, 0 being no confidence and 4 being high confidence in the finding. The degree of confidence was used as the vehicle for comparing the two groups of patients. RESULTS: The confidence ratings for all MRI variables were significantly higher for the zirconium group than for the cobalt chrome group. The confidence ratings varied less for the zirconium group than for the cobalt chrome group with an SD of 0.45 versus 0.95, respectively. There was greater interobserver agreement in the zirconium group (coefficient of interobserver agreement, 0.82 vs 0.35). The reviewers had the highest degree of confidence when examining for joint effusion in both groups (3.9 for the zirconium group; 3.7 for the cobalt chrome group). The greatest discrepancies between the two groups were in evaluation of periprosthetic osseous signal changes with the greatest difference being between the femoral component of each group with an average confidence rating of 3.3 for the zirconium group and 0.8 for the cobalt chrome group. CONCLUSION: Reviewers had significantly more confidence, less variability, and greater interobserver agreement in MRI evaluation of periprosthetic structures around zirconium knee prostheses than those around cobalt chrome knee prostheses. PMID- 16714673 TI - Reliability analysis of 16-MDCT in preoperative evaluation of total knee arthroplasty and comparison with intraoperative measurements. AB - OBJECTIVE: The aim of our study was to determine how consistent measurements of the distal femoral condyle on 16-MDCT are with intraoperative measurements and the reliability of 16-MDCT for the preoperative planning of total knee arthroplasty. MATERIALS AND METHODS: Between August 2003 and March 2004, 33 consecutive patients (two men and 31 women; age range, 53-89 years; mean age, 71 years) presenting with osteoarthritis underwent 16-MDCT (Mx 8000 IDT) of a total of 53 knees before total knee arthroplasty. The prospective analysis included measurements of transepicondylar distance, maximum anteroposterior dimension of medial and lateral femoral condyles, and trochlear width on a Rapidia workstation. To increase reliability, we repeated the measurements on a CT workstation after 2 months and compared them with the previous values. The values measured on the CT workstation were compared with the measurements obtained in the intraoperative field. Statistical analysis was performed using kappa statistics. A p value of less than 0.05 was considered statistically significant. RESULTS: The mean values of transepicondylar distance, maximum anteroposterior dimension of medial and lateral femoral condyles, and trochlear width were 75, 57, 58, and 38 mm at first measurement; 76, 58, 59, and 39 mm at second measurement on the CT workstation; and 79, 57, 60, and 42 mm at intraoperative measurement, respectively. At reliability analysis between the first measurements on the CT workstation and the intraoperative measurements, kappa values were 0.84 for the transepicondylar distance, 0.81 for the maximum anteroposterior dimension of the medial femoral condyle, 0.89 for the maximum anteroposterior dimension of the lateral femoral condyle, and 0.62 for the trochlear width. The kappa values for the second measurements were 0.86, 0.77, 0.84, and 0.61, respectively. Intraoperative measurements and measured values on the CT workstation showed excellent and almost perfect agreement, and intraobserver agreement was almost perfect. CONCLUSION: Femoral sizing on a CT workstation had excellent or almost perfect correlation with intraoperative measurements. Therefore, reliable preoperative planning for total knee arthroplasty may be done with 16-MDCT and an advanced workstation. PMID- 16714674 TI - Flare response in 18F-fluoride ion PET bone scanning. PMID- 16714675 TI - Sonography of brachial plexus traction injuries. AB - OBJECTIVE: Our purpose was to evaluate the feasibility of sonography in identifying nerve abnormalities in patients with traction injury of the brachial plexus. CONCLUSION: Sonography of the brachial plexus was technically feasible, although the entire brachial plexus could not be evaluated. Sonography appears to be a useful bedside imaging technique for assessing brachial plexus injury. The potential of sonography as a complementary diagnostic tool in the evaluation of these patients warrants further investigation. PMID- 16714676 TI - Internet-based dissemination of educational audiocasts: a primer in podcasting- how to do it. AB - OBJECTIVE: "Podcasting" is a relatively new Internet-based broadcast technology with unique characteristics that may prove valuable in radiologic education. In this article, we describe the procedure for generating, distributing, and downloading a podcast. CONCLUSION: Podcast technology holds promise as an inexpensive method to rapidly and cost effectively disseminate educational Internet audio broadcasts ("audiocasts"). PMID- 16714677 TI - On the AJR viewbox. Monomelic spread of metastatic disease due to proximal deep venous thrombosis. PMID- 16714678 TI - On the AJR viewbox. Pseudotumoral colonic tuberculosis complicating rheumatoid arthritis treated with a tumor necrosis factor antagonist. PMID- 16714679 TI - On the AJR viewbox. Pleuropulmonary endometriosis: CT-pathologic correlation. PMID- 16714680 TI - Hydatid cyst of the right atrium: a rare presentation. AB - Hydatid cyst of the heart is an uncommon lesion. Reported is the case of a 24 year-old female patient admitted with a history of chest pain and hemoptysis, and a mass lesion in the right atrium. Investigations and intraoperative intervention revealed the mass to be a hydatid cyst located in the right atrium, along with hydatid cysts of the liver and both lungs. The right atrial mass was successfully removed under cardiopulmonary bypass. PMID- 16714681 TI - Adult dysphagia associated with type B double aortic arch. AB - Esophageal compression secondary to a double aortic arch is rare in adults due to its propensity to cause severe symptoms in infancy or childhood. We report four adult patients with dysphagia associated with a double aortic arch. Chromosome analysis in one of these patients revealed a balanced translocation between chromosomes 4 and 18 - this has not been previously reported. Surgical division is indicated in all symptomatic patients and provides excellent operative results. PMID- 16714682 TI - Transbronchial localization of small pulmonary lesions for thoracoscopic resection. AB - The localization of small lung masses at thoracoscopic operation is very difficult. A 67-year-old female with tiny pulmonary metastases of renal cell carcinoma primary was successfully treated by pulmonary thoracoscopic resection after transbronchial localization using a dye. PMID- 16714683 TI - Active cerebral perfusion during carotid endarterectomy. AB - A patient with critical stenoses in the bilateral internal carotid arteries (ICA) required multivessel coronary revascularization. The diameter of the left ICA was far greater than the right, which strongly suggested that the cerebral circulation was highly dependent on the left. During left ICA endarterectomy, active cerebral perfusion of 300 mL x min(-1) at 23 degrees C using an extracorporeal circulation was employed through the ICA under repair. Subsequently, coronary bypass was performed on-pump with the heart beating. The postoperative course was very good. PMID- 16714684 TI - Papillary fibroblastoma of tricuspid valve with pulmonary embolization. AB - A 53-year-old man was diagnosed with a papillary fibroelastoma of the tricuspid valve with pulmonary embolization and associated coronary artery disease. He underwent excision of the tumor and coronary bypass grafting. Due to potential embolization, it is recommended that all such fibroelastomas be excised. PMID- 16714685 TI - Intraoperative aortic dissection in pediatric heart surgery. AB - Intraoperative aortic dissection occurred in a 3-year-old-boy undergoing repair of an atrial septal defect. Transesophageal echocardiography was useful for the diagnosis, and conservative medical treatment under close observation was feasible in this case which involved a limited intimal tear. PMID- 16714686 TI - Mitral valve papillary fibroelastoma: surgical considerations. AB - Cardiac papillary fibroelastomas are uncommon primary cardiac tumors. Due to their infrequent occurrence, an appropriate management strategy has been difficult to define. We report a 51-year-old female with a papillary fibroelastoma of the mitral valve diagnosed during a source of embolism evaluation, where the association between symptoms and the tumor were equivocal. The mitral valve papillary fibroelastoma was electively surgically excised, based on the proposed guidelines that allow for identification of patients at greatest risk of embolic events. We review the evaluation and selection of those patients who might benefit from surgical therapy even in the absence of symptoms. PMID- 16714687 TI - Post-pleuropneumonectomy herniation of liver mimicking major pulmonary embolism. AB - Following right-sided pneumonectomy and hemidiaphragm resection in a 58-year-old man with epithelioid mesothelioma, acute respiratory insufficiency and life threatening circulatory collapse developed after a forced Valsalva maneuver. Major pulmonary embolism was diagnosed on clinical grounds, however computed tomography revealed herniation of the liver into the right hemithorax. PMID- 16714688 TI - Chylothorax after aortic valve replacement. AB - A 71-year-old-man was diagnosed with chylothorax after aortic valve replacement. He was treated with a low-fat diet and pleural drainage with thoracentesis. The pleural effusion completely resolved by the 14th postoperative day. PMID- 16714689 TI - Transfusion related acute lung injury in cardiac surgery. AB - Transfusion related acute lung injury (TRALI) is an uncommon complication following administration of blood products. It is often difficult to differentiate from other commoner causes of cardio-respiratory instability. However, prompt diagnosis and management is associated with favorable outcome. PMID- 16714690 TI - Quality assurance in cardiac surgery: implications for Asia. PMID- 16714691 TI - Non-infectious aortitis: an important cause of severe aortic regurgitation. AB - From 1995 to 2004, 13 patients with severe aortic regurgitation due to non infectious aortitis underwent aortic valve surgery at our center. Twenty-eight operations (18 aortic valve replacements and 10 Bentall procedures including 1 aortic root replacement with a homograft) were performed due to a high incidence of prosthetic valve or valved conduit dehiscence (54%). Steroid therapy reduced the incidence of re-operation for anastomotic dehiscence within one year from 90% to 18%. Strategies to prevent dehiscence of the prosthetic aortic valve, which include institution of immunosuppression, replacement of the diseased aorta, and monitoring inflammatory indices, are discussed. PMID- 16714692 TI - Experimental study of effect of Fontan circuit on pulmonary microcirculation. AB - Pulsatile pulmonary blood flow plays an important role in regulating shear-stress mediated release of endothelium-derived nitric oxide and endothelin-1, and it reduces pulmonary vascular resistance by passive capillary recruitment. The aim of this study was to demonstrate changes in pulmonary capillary structure and endothelial function induced by the chronic nonpulsatile flow of the Fontan circulation. A canine model with nonpulsatile flow in the right lung was established, and sacrificed 3 months later. Compared to the left lung, wall thickness of the pulmonary arterioles was thinner, endothelin-1 expression was weaker, endothelial nitric oxide synthase activity was stronger, and there was a good correlation between the histomorphometric and immunohistochemical findings. These data indicate that long-term nonpulsatile flow can lead to endothelial dysfunction, which may be involved in distention and vascular structure remodeling due to the increase in pulmonary vascular resistance; but it also can lead to increased patency of the arteriovenous shunt, which might be at least partly involved in pulmonary arteriovenous fistula development and exercise limitation after the Fontan operation. PMID- 16714694 TI - Optimizing early extubation after coronary surgery. AB - Early extubation after isolated coronary artery bypass surgery was assessed retrospectively in 545 of 779 patients treated by the same surgical team over one year. All underwent extubation within 10 hr of arrival at the cardiothoracic intensive care unit: 343 in < 6 hr and 202 in 6-10 hr. Operative mortality was 2.2%. Group comparisons revealed that patients who had earlier extubation were younger (61 vs. 66 years; p < 0.001), more likely to be male (72.5% vs. 61.3%; p < 0.05), with a shorter aortic crossclamp time (49.2 +/- 15.0 vs. 53.3 +/- 14.0 min; p < 0.05), cardiopulmonary bypass time (65 +/- 18.4 vs. 72.2 +/- 19.2 min; p < 0.05), intensive care unit stay (18.8 +/- 5.6 vs. 22.4 +/- 3.2 hr; p < 0.05) and postoperative hospital stay (5.2 +/- 2.2 vs. 6.0 +/- 2.4 days; p = 0.01). Extubation < 6 hr after cardiopulmonary bypass may accelerate recovery. The finding of no significant differences in clinical parameters between the groups suggests that efforts to further reduce the time to extubation might be worthwhile. PMID- 16714693 TI - Late mortality and morbidity in elderly patients with mechanical heart valves. AB - A retrospective study was performed in patients under and over 65 years old implanted with a mechanical valve, to compare late mortality and morbidity. Of 381 patients who underwent mechanical valve replacement at Nagoya University in the 1990s, 357 (11 hospital deaths and 13 lost to follow-up; 96.4% follow-up rate) were followed up for 7.9 +/- 3.3 years (2,811 patient-years). They were divided into two groups either side of 65 years of age at operation. The young and elderly patient groups contained 275 and 82 patients, respectively. The survival rate in the young group was 96.1% (95% confidence interval, 93.7%-98.5%) at 5 years and 92.0% (95% confidence interval, 88.3%-95.7%) at 10 years, which was significantly better than 88.0% (95% confidence interval, 80.6%-95.4%) at 5 years and 73.8% (95% confidence interval, 66.2%-85.4%) at 10 years in the elderly group. The two groups did not differ significantly in the incidence of thromboembolic events, bleeding events, endocarditis, or reoperation. We are also encouraged by the fact that mechanical valves are not a risk factor for late mortality or morbidity, even in elderly patients. PMID- 16714695 TI - 15-year comparison of supra-annular porcine and PERIMOUNT aortic bioprostheses. AB - The second-generation Carpentier-Edwards bioprostheses, the supra-annular porcine valve and the PERIMOUNT pericardial valve, have been evaluated longitudinally for several years. This study compared clinical performance over 15 years. Aortic valve replacement was performed with a supra-annular porcine valve in 1,823 patients (group 1) aged 19-89 years (mean, 68.9 +/- 10.9 years) and with a PERIMOUNT pericardial bioprosthesis in 1,430 patients (group 2) aged 16-90 years (mean, 69.5 +/- 10.4 years). The groups were similar except for concomitant coronary artery bypass in 43% of group 1 and 18% of group 2 ( p < 0.001). Overall survival at 15 years was 29.3% +/- 1.5% for group 1 and 35.2% +/- 3.1% for group 2 ( p = 0.0009). The actual freedom from valve-related mortality was 88.5% +/- 0.9% for group 1 and 84.9% +/- 1.7% for group 2. The actual freedom from structural valve deterioration at 15 years was similar overall, and for patients aged > 60 years, between the groups, but was dissimilar (group 2 > group 1) for age group 2), sex (male > female), age, and concomitant coronary artery bypass. Both bioprostheses provided satisfactory clinical performance at 15 years after aortic valve replacement. PMID- 16714696 TI - Management of malignant mesothelioma by decortication and adjunct phototherapy. AB - Malignant mesothelioma is a relatively rare tumor that originates in the pleural space and almost invariably results from exposure to asbestos. Between September 1989 and December 1999, 100 patients were managed with curative intent using a combination of full decortication, adjunct phototherapy after administration of hematoporphyrin derivative, and strip radiotherapy to any areas where adequate clearance was not obtained. The survival curve was compared to that of 17 matched patients treated by decortication alone. Median survival increased from 250 to 440 days in the combined treatment group. PMID- 16714697 TI - Clinical characteristics of bicuspid aortic valves in surgical patients. AB - Of 413 patients undergoing surgery for aortic valve disease, 42 were confirmed to have congenital bicuspid aortic valve. There were 24 males and 18 females, with a mean age of 54.9 +/- 14.6 years. The etiology of the surgical indications in patients with bicuspid valves were degeneration, rheumatic disease, and infectious endocarditis. Thirty-two patients had aortic stenosis and 10 had aortic regurgitation. Infective endocarditis was seen mainly in patients < or =50 years of age at the time of surgery, while degenerative aortic stenosis occurred more often in older patients. The annular diameter measured during surgery was significantly narrower in the 42 patients with bicuspid valves compared to 371 patients with tricuspid valves who underwent aortic valve replacement during the same period. It is important to prevent infectious endocarditis in younger patients diagnosed with bicuspid aortic valve, whereas careful attention should be paid to decalcification during surgery in older patients because the vast majority suffer from degenerative aortic stenosis. PMID- 16714698 TI - Right ventricular dysfunction due to right ventricular outflow tract patch. AB - Doppler tissue imaging analysis was used to examine the relationship between right ventricular function and right ventricular outflow tract damage in 54 patients with repaired tetralogy of Fallot. The patients were divided into three groups: 16 in whom the right ventricular outflow tract was directly sutured (group DS), 23 who had transventricular patch repair (group TVP), and 15 who had transannular patch repair (group TAP). The control group consisted of 16 age matched patients who underwent patch closure of a ventricular septal defect (group C). The Tei index was obtained from tricuspid and pulmonary Doppler flow velocities. The right ventricular Tei index was significantly greater in groups TVP and TAP than in group DS. Doppler tissue imaging analysis in groups TVP and TAP showed shorter myocardial systolic velocity, diastolic peak velocity, and atrial diastolic peak velocity, lower peak myocardial velocity and acceleration during isovolumic contraction, and prolonged isovolumic relaxation and contraction times compared to groups DS and C. Right ventricular dysfunction is due to the right ventricular outflow tract patch. Thus, the right ventricular outflow tract may be essential for right ventricular ejection and maintenance of right ventricular function. PMID- 16714699 TI - Surgical management of bilateral bronchiectases: results in 29 patients. AB - Bronchiectasis is a major cause of morbidity and mortality in developing countries. Staged bilateral segmental resection of the lungs is performed in selected patients. Our experience of surgical removal of 87 bilateral bronchiectases in 29 patients during an 11-year period was reviewed retrospectively. High-resolution computed tomography was performed preoperatively in all patients to locate the anatomic sites of bronchiectasis. The mortality and morbidity of the surgical procedure, clinical symptoms, age distribution, etiology, bacteriology, and operative procedures were analyzed. There were 22 males (76%) and 7 females (24%), aged 5 to 60 years, with a mean age of 30 years. Complications developed in 11 patients (38%); atelectasia was the most common (14%). There was one hospital death. Clinical symptoms disappeared in 19 (66%) patients, improved in 5 (17%), and were unchanged in 4 (14%). Staged bilateral resection for bronchiectases can be performed at any age with acceptable morbidity and mortality. PMID- 16714700 TI - Experimental assessment of the drainage capacity of small silastic chest drains. AB - Recently, flexible fluted small silicone drains have been used widely as chest drains after cardiac surgery. Despite the clinical advantages of using smaller silastic chest drains over conventional chest tubes, an experimental comparison of the drainage capacity between these two drain tubes has not yet been performed. The drainage capacity of 19F silicone drains and 28F conventional tubes was tested. In an in vitro study, both tubes were set in a water bath and drained at a pressure of 10 mm Hg. In an in vivo study, the drains were inserted into the hemithorax in 8 adult pigs. Blood was infused at 20 mL x min(-1) into both chest cavities and the tubes were drained at 15 cm H(2)O for 30 min. In the in vitro study, the drainage capacity of the conventional chest tube was 9-times higher than that of the smaller silicone drain (103.8 vs. 11.6 L x hr (-1)). However, in the in vivo study, there was no difference in drainage capacity between the two different tubes over time. This experiment demonstrated that the smaller silastic chest drain has sufficient drainage capacity, almost identical to the conventional chest tube, in the clinical setting. PMID- 16714701 TI - Off-pump coronary endarterectomy in high-risk patients. AB - The outcome of off-pump endarterectomy in patients with diffuse coronary artery disease and increased risk factors for cardiopulmonary bypass was reviewed. Thirty-eight procedures were carried out in 32 patients (21 men and 11 women) aged 59 to 78 years (mean, 69 years). Mean left ventricular ejection fraction was 38.6% (range, 24% - 55%). The number of grafts per patient was 2.6. The left internal mammary artery was used in 29 patients (91%). Endarterectomy was performed on the right coronary artery and its branches (18), the left anterior descending artery (15), diagonal branch (2), and the circumflex system (3). Overall operative mortality was 3.1%. Perioperative myocardial infarction occurred in 6.2%. Follow-up was complete in 30 patients (94%). The mean follow-up period was 14 +/- 3.3 months. Late survival was 93%. Freedom from cardiac events that required hospital re-admission was 89%. Freedom from angioplasty of the endarterectomized vessel was 96%. These findings indicate that off-pump endarterectomy can be performed with good results in patients with diffuse coronary artery disease. PMID- 16714702 TI - Left ventricular aneurysmectomy: tailored scar excision and linear closure. AB - Coronary artery bypass surgery with or without aneurysmectomy is used to treat patients with left ventricular aneurysm. There is debate about patient selection and the appropriate surgical technique. We analyzed the results of 102 consecutive patients who underwent left ventricular aneurysmectomy and reconstruction using a modified linear closure technique between 1992 and 2003. The mean age was 62 years, 81% of the patients were male, and 47% had an ejection fraction < 35%. The locations of the left ventricular aneurysms were anteroapical (75%), apical (21%), and posteroinferior (4%); 23% contained thrombi. Additional procedures included aortic valve replacement in 4, mitral valve repair in 1, and coronary bypass grafting in 98 patients; 3 underwent isolated repair of left ventricular aneurysm. Hospital mortality was 7% and long-term survival was 76% at a mean follow-up of 39 months. Most patients improved symptomatically postoperatively. Left ventricular aneurysm repair with tailored scar excision and a modified closure technique is associated with acceptable mortality and long term survival. PMID- 16714703 TI - Outcome of Glenn anastomosis for heterotaxy syndrome with single ventricle. AB - A retrospective study was undertaken of 157 patients who underwent a Glenn anastomosis between January 1996 and May 2001. Of these, 33 had heterotaxy syndrome: 20 males and 13 females, with a mean age of 1.26 +/- 2.8 years. Twenty five had right atrial isomerism and 5 had left isomerism. A common atrioventricular valve was found in 24 patients, 18 had bilateral superior venae cavae, and 18 had anomalous pulmonary venous return. Repair was carried out in 8 patients with anomalous pulmonary venous return, and pulmonary artery augmentation was performed in 11. Compared to the 124 patients who had a Glenn operation for single ventricle without heterotaxy, there were significantly longer durations of mechanical ventilation, intensive care unit stay, and inotropic support, as well as higher mortality in the heterotaxy group. Heterotaxy syndrome with single ventricle still has a high rate of morbidity and mortality. Patients with severe atrioventricular valve regurgitation are at risk of early death. Complete Fontan circulation may not be possible in all patients, and Glenn anastomosis may be their final palliation. PMID- 16714704 TI - Landiolol enhances effect of ischemic preconditioning in isolated rabbit hearts. AB - The effect of the ultra-short-acting beta blocker, landiolol, on ischemic preconditioning was examined in isolated rabbit hearts. Ischemic preconditioned hearts received 2 episodes of 5 min each of global ischemia and reperfusion. The left anterior descending coronary artery was occluded for 1 hour and reperfused for 1 hour. Left ventricular end-systolic and end-diastolic pressures and infarct size were measured. Seven control hearts had no drug infused. Four groups of 6 hearts each were pretreated with 1 or 3 microM of landiolol or a combination of 1 or 3 microM landiolol and ischemic preconditioning. A further group of 6 hearts had ischemic preconditioning without landiolol. Ischemic preconditioning significantly reduced left ventricular end-diastolic pressure and infarct size compared to the controls. Landiolol alone did not change left ventricular end diastolic pressure or infarct size, but landiolol 3 microM and ischemic preconditioning decreased left ventricular end-diastolic pressure more than preconditioning alone. These data suggest that pre-ischemic landiolol infusion may enhance the cardioprotective effect of ischemic preconditioning. PMID- 16714705 TI - Giant bronchogenic cyst mimicking tension pneumothorax. AB - An 18-month-old girl presented with high fever and vomiting. Pneumothorax and a cystic formation in the right hemithorax were found on a chest radiograph. The cyst measuring 10 x 10 x 8 cm was resected by a simple wedge resection. Histology revealed a complicated bronchogenic cyst with abscess formation. PMID- 16714706 TI - Graft replacement and muscle wrap for infected aneurysm of thoracic aorta. AB - A 68-year-old man was admitted with high fever and enlargement of the distal aortic arch on chest radiography. Computed tomography showed a huge proximal descending thoracic aortic aneurysm. Graft replacement of the proximal descending thoracic aorta was performed on an emergency basis, and a pectoralis major muscle flap was wrapped around the graft. Salmonella enteritidis was detected in the resected tissue. The patient recovered well, with no signs of infection. PMID- 16714707 TI - Combined minimally invasive coronary bypass surgery and left pulmonary lobectomy. AB - Two patients underwent combined heart and lung surgery performed through a limited left anterior thoracotomy. Good exposure of the left pulmonary hilum and the left anterior descending artery was obtained, allowing left upper lobectomy combined with off-pump coronary bypass grafting. This approach avoids potential complications due to sternotomy, staged operations, and cardiopulmonary bypass. These cases show that a limited thoracotomy allows safe and effective combined lung and cardiac surgery in carefully selected patients. PMID- 16714708 TI - Mitral valve repair on the beating perfused heart. AB - It is difficult to assess the success of mitral valve repair in the arrested heart. Various techniques have been described. Transesophageal echocardiogram (TEE) provides excellent two-dimensional evaluation of the repair, but three dimensional anatomic characteristics are limited. We describe a simple technique for performing mitral valve repair on the beating heart. This allows accurate evaluation of valvular competence and three-dimensional anatomic characteristics prior to closure of the atriotomy. PMID- 16714709 TI - Pathobiology of idiopathic ascending aortic aneurysms. AB - The majority of ascending aortic aneurysms cannot be related to any specific etiology and should be qualified as idiopathic. The pathobiology of ascending aortic aneurysms remains incompletely understood. Data from direct study are still scarce and often limited because of patient heterogenicity. Currently available information suggests that destructive remodeling of the aortic wall, inflammation and angiogenesis, biomechanical wall stress, and molecular genetics are relevant mechanisms of idiopathic ascending aortic aneurysm formation and progression. Further understanding of these mechanisms will likely provide novel diagnostic, prognostic, and therapeutical tools for the clinician. PMID- 16714710 TI - Giant chondroblastic osteosarcoma of the chest wall. PMID- 16714711 TI - Congenital kinking of aortic arch associated with thoracic aortic aneurysm. PMID- 16714712 TI - Mechanical valves and pregnancy: a caveat. PMID- 16714714 TI - Influenza virus associated encephalopathy. PMID- 16714716 TI - Balancing biomedical, care, and support needs in the technology dependent child. PMID- 16714715 TI - The great weight gain experiment, accelerators, and their implications for autoantibodies in diabetes. PMID- 16714717 TI - Parsing ketotic hypoglycaemia. PMID- 16714718 TI - Prehospital management of the acutely ill child. PMID- 16714721 TI - Placental malaria and immunity to infant measles. AB - The efficiency of transplacental transfer of measles specific antibody was assessed in relation to placental malaria. Infection at delivery was associated with a 30% decrease in expected cord measles antibody titres. Uninfected women who received anti-malarial drugs during pregnancy transmitted 30% more antibody than those who received no antimalarial drugs. PMID- 16714722 TI - Cullen's sign and massive ovarian enlargement secondary to primary hypothyroidism in a patient with a normal FSH receptor. AB - Ovarian hyperstimulation is a recognised complication of longstanding hypothyroidism. A 12 year old girl with atrophic thyroiditis who presented with abdominal pain and distension is reported. She was noted to have bruising in the vicinity of the umbilicus (Cullen's sign). She had pronounced ovarian enlargement on ultrasonography and it was hypothesised that this profound phenotype might reflect an abnormal FSH receptor. However sequencing of the FSH receptor was normal. The ovarian enlargement resolved with thyroxine replacement. Physicians and surgeons should consider longstanding hypothyroidism in patients presenting with Cullen's sign. PMID- 16714723 TI - Community associated MRSA: an alert to paediatricians. AB - Community associated methicillin resistant Staphylococcus aureus (CA-MRSA) is an emerging pathogen typically associated with skin and soft tissue infection, with occasional reports of fatality in previously healthy children and young adults. We report a case of invasive CA-MRSA and highlight the potential impact of such infections on empirical treatment of staphylococcal infections. PMID- 16714724 TI - Tuberculous pyomyositis of the left quadratus lumborum. PMID- 16714726 TI - Congenital subependymal giant cell astrocytoma diagnosed on fetal MRI. PMID- 16714725 TI - Optimising management in Turner syndrome: from infancy to adult transfer. AB - Turner syndrome can be defined as loss or abnormality of the second X chromosome in at least one cell line in a phenotypic female. The condition occurs in approximately 1 in every 2000 live female births,(1) so that in the UK the prevalence for any year of life is in the region of 200 girls. The condition is much more common in utero, it being estimated that 1-2% of all conceptuses are affected, of whom only 1% will survive to term. PMID- 16714727 TI - Shwachman-Diamond syndrome: UK perspective. AB - So much has been added to our knowledge of Shwachman-Diamond syndrome (SDS) since it was last reviewed in this journal some 25 years ago, that there is now an urgent need to bring the condition to the attention of a new generation of paediatricians. SDS, although a rare autosomal recessive disorder, demands wide attention because it features in the differential diagnosis of a number of important childhood diseases. It can be diagnosed in children of all ages, or in adults. SDS most commonly presents in infancy with features of exocrine pancreatic insufficiency, bone marrow dysfunction, and short stature. PMID- 16714728 TI - NICE guidelines and the epilepsies: how should practice change? AB - The care and provision for children and adults with epilepsy and their carers has recently been under scrutiny with a series of reports highlighting concerns and calling for change. The National Institute for Clinical Excellence (NICE) published recommendations for management of adults and children in October 2004. Although recommendations were often specific and practical they did not include precise details regarding their implementation. Key recommendations and their implications are discussed in this review. PMID- 16714730 TI - Towards evidence based medicine for paediatricians. PMID- 16714729 TI - Does BCG have a role in tuberculosis control and prevention in the United Kingdom? AB - The United Kingdom has recently changed its BCG vaccination policy in response to the changing epidemiology of tuberculosis (TB) in children. One of the changes has been the abandonment of the long standing school's BCG programme because of the low risk of TB in that population. The other change has been the targeting of those infants and children at increased risk of TB, particularly in populations with increased rates of TB. However, there remain questions as to what role BCG plays in TB control and prevention in the UK. PMID- 16714731 TI - How much regret are you prepared to take? PMID- 16714732 TI - How useful is C-reactive protein in detecting occult bacterial infection in young children with fever without apparent focus? PMID- 16714733 TI - Does treatment with Echinacea purpurea effectively shorten the course of upper respiratory tract infections in children? PMID- 16714734 TI - In juvenile idiopathic arthritis, is folate supplementation effective against methotrexate toxicity at the expense of methotrexate's efficacy? PMID- 16714735 TI - Ankyloglossia intervention in outpatients is safe: our experience. PMID- 16714736 TI - Best practice: migraine. PMID- 16714737 TI - Postgraduate training in the UK: whither uniformity? PMID- 16714738 TI - Tongue ties and breast feeding * Authors' reply. PMID- 16714739 TI - Exercise induced dyspnoea: if not asthma, then what? PMID- 16714740 TI - Breast feeding method should ensure rapid weight gain. PMID- 16714741 TI - Antenatally diagnosed single kidney: lack of uniformity in postnatal management practice. PMID- 16714742 TI - Buccal midazolam: is a test dose in hospital needed? PMID- 16714743 TI - Accident and emergency: a gateway to improve the management of atopic disease. PMID- 16714744 TI - NICE guidelines for hyperlipidaemia in children and young people with type I diabetes: time for a rethink? PMID- 16714745 TI - Three-dimensional MRI of the male urethrae with implanted artificial sphincters: initial results. AB - The aim of this study was to develop a method for simultaneous 3D visualization of a new type of artificial urethral sphincter (AUS) and adjacent urinary structures. Serial MR tomograms were acquired from seven men after AUS implantation. 3D reconstruction was performed by thresholding original (positive) and inverted (negative) image intensity and by subsequently fusing positive and negative images. Results show that the bladder, cuff and balloons of the AUS of originally high intensity were imaged in 3D by thresholding the positive datasets. The urethrae and corpora cavernosa penis of originally low intensity were displayed in 3D by thresholding the negative datasets. Fusion of the positive and negative datasets allowed simultaneous visualization of the AUS complex and adjacent urinary structures. All the structures of interest were also clearly seen by interactive multiplanar reformatting. Coronal tomographic datasets provided better 3D and reformatted 2D images than sagittal and transverse datasets. This technique offers a simple means for evaluating the complex urethral anatomy and the AUS, and has potential for improved 3D visualization of many other complex morphological and pathological conditions. PMID- 16714746 TI - Heel bone densitometry: device specific thresholds for the assessment of osteoporosis. AB - A diagnosis of osteoporosis is facilitated by bone mineral density (BMD) measurement of the lumbar spine and hip using dual energy X-ray absorptiometry (DXA), interpreted in accordance with criteria published by the World Health Organization (WHO). The use of peripheral DXA is growing in primary care and guidance on its use has recently been published by the National Osteoporosis Society (NOS), recommending a triage approach using thresholds specific to each type of peripheral device. However, no data currently exist for the Norland Apollo heel densitometer (Cooper Surgical, Trumbull, USA). 215 women between 50 years and 75 years of age (mean age 64.6 years) referred for hip and spine BMD measurements also had a heel BMD measurement. Device specific upper and lower thresholds were calculated for the Norland Apollo heel densitometer to give a 90% sensitivity and 90% specificity for osteoporosis at the hip or spine. Patients with a heel T-score of above -1.2 are very likely to have normal bone density on axial densitometry, whilst patients with heel T-score of below -2.2 are very likely to have osteoporosis at the hip or spine. Patients whose measurements lie between the thresholds should be referred for axial DXA. PMID- 16714747 TI - The impact of a short course of study on the performance of radiographers when highlighting fractures on trauma radiographs: "The Red Dot System". AB - This study was set up to determine the effect of a short course of study on radiographers' ability to identify fractures in a trauma context (Red Dot system). The performance of 133 radiographers attending Red Dot courses delivered from April 1999 to September 2003 was evaluated prospectively using a selected sample of radiographs. Assessment of fracture identification occurred before, immediately after and 6 months after the course. The same film set was used and consisted of 30 axial and appendicular cases, 18 with fractures and 12 normal or normal variants. Following a test for normality of the data, a Wilcoxon Signed Rank Test was selected and paired tests were done between each assessment for sensitivity and specificity. There were significant differences in sensitivity between all three assessments (por=100% of the dose. Compared with AP/PA and 3-field plans, IMRT plans had a greater percentage of target receiving the prescribed dose, but also a greater volume receiving >110% of the dose. IMRT was well tolerated; no patients developed acute gastrointestinal toxicity greater than grade 2. All seven experienced grade 2 nausea, three had grade 2 diarrhoea and two had grade 2 oesophagitis. Weight loss ranged from 0-12% (mean 6.1% and median 5.8%). IMRT in the treatment of gastric malignancies reduces the mean and above threshold doses to critical normal tissues. In an initial cohort of seven patients, 50.4 Gy delivered by IMRT is well tolerated and safe. PMID- 16714753 TI - Changes in applicator positions and dose distribution between high dose rate brachytherapy fractions in cervix carcinoma patients receiving definitive radiotherapy. AB - This study examines the change of applicator geometry and its effect on rectal/rectum (R) and bladder (B) doses, and obtained radiobiological equivalent doses (RED), between each high dose rate (HDR) brachytherapy (BT) fraction in cervical carcinoma patients. BT using a tandem (T) and two ovoids (O) is included, and any discrepancies in applicator positions among the fractions were calculated. Whether the change of applicator position had an effect on the calculated R and B doses was analysed. Furthermore, the relationship between the size of tumour, the magnitude of displacement and the change in R and B doses was also investigated. Lastly, the changes in R and B RED were noted. The average magnitude of displacement was between 2.0 mm and 16.9 mm, showing time trend. There was no relationship between tumour size and the magnitude of discrepancy of Left O, Right O, T, R, B, and neither change in R and B doses (p>0.05). The mean differences of R and B doses were between 49-78 cGy, and 70-84 cGy, respectively. The magnitude of discrepancy and changes in doses showed no correlation (p>0.05). There were no significant differences in REDs for bladder (p = 0.8) and rectum (p = 0.2). In conclusion, there were significant differences in the applicator positions R and B and R and B doses among the fractions, which confirm the necessity of treatment planning in each HDR BT fraction. However, the total calculated R and B REDs did not show a remarkable difference. PMID- 16714754 TI - Clinical and cellular ionizing radiation sensitivity in a patient with xeroderma pigmentosum. AB - XP14BR is a cell line derived from a xeroderma pigmentosum (XP) patient from complementation group C. The patient was unusual in presenting with an angiosarcoma of the scalp, treated by surgical excision and radiotherapy. Following 38 Gy in 19 fractions with 6 MEV electrons, a severe desquamation and necrosis of the underlying bone ensued, and death followed 4 years later. The cell line was correspondingly hypersensitive to the lethal effects of gamma irradiation. We had previously shown that this sensitivity could be discriminated from that seen in ataxia-telangiectasia (A-T). The cellular response to ultraviolet radiation below 280 nm (UVC) was characteristic of XP cells, indicating the second instance, in our experience, of dual cellular UVC and ionizing radiation hypersensitivity in XP. We then set out to evaluate any defects in repair of ionizing radiation damage and to verify any direct contribution of the XPC gene. The cells were defective in repair of a fraction of double strand breaks, with a pattern reminiscent of A-T. The cell line was immortalized with the vector pSV3neo and the XPC cDNA transfected in to correct the defect. The progeny derived from this transfection showed the presence of the XPC gene product, as measured by immunoblotting. A considerable restoration of normal UVC, but not ionizing radiation, sensitivity was observed amongst the clones. This differential correction of cellular sensitivity is strong evidence for the presence of a defective radiosensitivity gene, distinct from XPC, which is responsible for the clinical hypersensitivity to ionizing radiation. It is important to resolve how widespread ionizing radiation sensitivity is amongst XP patients. PMID- 16714755 TI - Survival and initial chromatid breakage in normal and tumour cells exposed in vitro to gamma rays and carbon ions at the HIRFL. AB - Human hepatoma and normal liver cells were irradiated with (12)C(6+) ion beams (linear energy transfer (LET) = 96 keV microm(-1)) and gamma-rays at the Heavy Ion Research Facility in Lanzhou (HIRFL). The numbers and types of chromatid breaks were detected using the premature chromosome condensation technique. Irradiation with (12)C(6+) ions produced a majority of isochromatid break types, while chromatid breaks were dominant for irradiation with gamma-rays. Experimental results showed that the initial level of chromatid breaks is clearly related to the absorbed dose from (12)C(6+) ions and gamma-rays. The (12)C(6+) ions are relatively more effective at inducing initial chromatid breaks when compared with the gamma-rays. A relative biological effectiveness (RBE) of about 2.5 resulted for the induction of initial chromatid breaks by (12)C(6+) ions relative to gamma-rays in both cell lines. PMID- 16714756 TI - Usefulness of mini-tracheostomy and torque controlled insertion of applicator in fractionated endobronchial brachytherapy. AB - Endobronchial brachytherapy was developed as effective treatment of endobronchial cancer and fractionated schedule is applied to decrease late toxicity. However, repeated bronchofiberscopy is onerous to the patient and restricts the treatment schedule itself. We applied mini-tracheostomy for a ready access route, and a torque controlled technique for easy insertion of the endobronchial applicator. Eight patients with tracheobronchial cancer invasion were treated with endobronchial brachytherapy of 18-30 Gy/3-5 fractions/1.5-2.5 weeks (median 24 Gy/4 fractions/2 weeks) at reference points of 5 mm from the bronchial surface. The averaged individual irradiation and single session times were 4 min and 24 min, respectively. There were no procedure-related complications. These technical improvements may facilitate flexible fractionated dose prescriptions. PMID- 16714757 TI - MR findings of penile lymphoma. AB - Penile lymphoma is extremely rare and secondary involvement of the penis by lymphoma may be due to retrograde spread or to direct extension from neighbouring organ. The appearance of penile lymphoma varies and can be mistaken for other soft tissue tumours. We report on a case with malignant lymphoma of the penis. MRI findings revealed soft-tissue mass of homogeneous isointensity around the middle to distal part of penis on T(1) weighted imaging and T(2) weighted imaging. It was well encapsulated, minimally enhanced and distinct from corpus cavernosum and corpus spongiosum. PMID- 16714758 TI - Deep tracheal laceration after balloon dilation for benign tracheobronchial stenosis: case reports of two patients. AB - We report two cases of deep tracheal laceration in female patients after balloon dilation for benign tracheobronchial stenosis. Immediate post-procedure bronchoscopy and CT including 3D reconstructions showed deep lacerations in the posterior tracheal wall. Clinically, the patients' dyspnoea subsided and there has been no recurrence during follow-up after balloon dilation. On the follow-up 3D-reconstructed CT scans obtained 2 months and 8 months following balloon dilation, respectively, the lacerations had healed completely and there was considerable improvement in lumen size. PMID- 16714760 TI - Any fool can understand IMRT. PMID- 16714761 TI - Neck pain: an unusual presentation of a common disease. PMID- 16714762 TI - Mass spectrometric identification of human prostate cancer-derived proteins in serum of xenograft-bearing mice. AB - Lack of sensitivity and specificity of current tumor markers has intensified research efforts to find new biomarkers. The identification of potential tumor markers in human body fluids is hampered by large variability and complexity of both control and patient samples, laborious biochemical analyses, and the fact that the identified proteins are unlikely produced by the diseased cells but are due to secondary body defense mechanisms. In a new approach presented here, we eliminate these problems by performing proteomic analysis in a prostate cancer xenograft model in which human prostate cancer cells form a tumor in an immune incompetent nude mouse. Using this concept, proteins present in mouse serum that can be identified as human will, by definition, originate from the human prostate cancer xenograft and might have potential diagnostic and prognostic value. Using one-dimensional gel electrophoresis, liquid chromatography, and mass spectrometry, we identified tumor-derived human nm23/nucleoside-diphosphate kinase (NME) in the serum of a nude mouse bearing the androgen-independent human prostate cancer xenograft PC339. NME is known to be involved in the metastatic potential of several tumor cells, including prostate cancer cells. Furthermore we identified six human enzymes involved in glycolysis (fructose-bisphosphate aldolase A, triose-phosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, alpha enolase, and lactate dehydrogenases A and B) in the serum of the tumor bearing mice. The presence of human NME and glyceraldehyde-3-phosphate dehydrogenase in the serum of PC339-bearing mice was confirmed by Western blotting. Although the putative usefulness of these proteins in predicting prognosis of prostate cancer remains to be determined, the present data illustrate that our approach is a promising tool for the focused discovery of new prostate cancer biomarkers. PMID- 16714764 TI - The biochemical role of the heat shock protein 90 chaperone complex in establishing human telomerase activity. AB - Telomerase is a ribonucleoprotein complex that synthesizes the G-rich DNA found at the 3'-ends of linear chromosomes. Human telomerase consists minimally of a catalytic protein (hTERT) and a template-containing RNA (hTR), although other proteins are involved in regulating telomerase activity in vivo. Several chaperone proteins, including hsp90 and p23, have demonstrable roles in establishing telomerase activity both in vitro and in vivo, and previous reports indicate that hsp90 and p23 are required for the reconstitution of telomerase activity from recombinant hTERT and hTR. Here we report that hTERT and hTR associate in the absence of a functional hsp90-p23 heterocomplex. We also report that hsp90 inhibitors geldanamycin and novobiocin inhibit recombinant telomerase even after telomerase is assembled. Inhibition by geldanamycin could be overcome by allowing telomerase to first bind its primer, suggesting a role for hsp90 in loading telomerase onto the telomere. Inhibition by novobiocin could not similarly be overcome but instead resulted in destabilization of the hTERT polypeptide. We propose that the hsp90-p23 complex fine tunes and stabilizes a functional telomerase structure, allowing primer loading and extension. PMID- 16714763 TI - Crucial roles of Sp1 and epigenetic modifications in the regulation of the CLDN4 promoter in ovarian cancer cells. AB - Claudins form a large family of tight junction proteins that have essential roles in the control of paracellular ion flux and the maintenance of cell polarity. Many studies have shown that several claudin family members are abnormally expressed in various cancers. In particular, CLDN4 (encoding claudin-4) is overexpressed in ovarian cancer. However, although CLDN4 overexpression is well established, the mechanisms responsible for this abnormal regulation remain unknown. In the present study, we delineate a small region of the CLDN4 promoter critical for its expression. This region contains two Sp1 sites, both of which are required for promoter activity. However, because of the ubiquitous expression of Sp1, these sites, although necessary, are not sufficient to explain the patterns of gene expression of CLDN4 in various ovarian tissues. We show that the CLDN4 promoter is further controlled by epigenetic modifications of the Sp1 containing critical promoter region. Cells that overexpress CLDN4 exhibit low DNA methylation and high histone H3 acetylation of the critical CLDN4 promoter region, and the reverse is observed in cells that do not express CLDN4. Moreover, the CLDN4-negative cells can be induced to express CLDN4 through treatment with demethylating and/or acetylating agents. Because CLDN4 is elevated in a large fraction of ovarian cancer, the mechanism leading to deregulation may represent a general pathway in ovarian tumorigenesis and may lead to novel strategies for therapy and an overall better understanding of the biology of this disease. PMID- 16714765 TI - Lower mutagenicity but higher stability of Cr-DNA adducts formed during gradual chromate activation with ascorbate. AB - Recent epidemiological and risk assessment studies have found a very high risk of lung cancer among chromium(VI)-exposed workers even at permissible levels of exposure. However, mechanistic views on the key genotoxic role of transient Cr(V) intermediates were more consistent with the threshold or highly non-linear (heavy dose) models of genetic damage by intracellular Cr(VI). In this work, we examined the production of mutagenic DNA lesions during metabolism of Cr(VI) by its dominant reducer ascorbate (vitamin C) under conditions promoting increased yield of transient Cr forms. We found that slow reductive activation of Cr(VI) by limited concentrations of ascorbate resulted in a greater yield of DCFH-oxidizing Cr intermediates but these species were unable to cause DNA strand breaks. Cr(VI) ascorbate reactions generated a high number of Cr-DNA adducts that were responsible for all mutagenic responses detected in Cr(VI)-treated pSP189 shuttle plasmids following their replication in human cells. Mutagenicity of DNA damage resulting from the reactions with increased stability of Cr intermediates was approximately four times lower relative to the conditions lacking detectable Cr(V) formation. Unlike other reactions, slow reduction of Cr(VI) with ascorbate produced Cr-DNA adducts that were more resistant to dissociation by chelators, suggesting multicoordinate binding of Cr(III) to DNA. Overall, our findings do not support the possibility that increased Cr(V) formation at depleted ascorbate levels modeling heavy dose exposures causes higher levels of mutagenic DNA damage. PMID- 16714766 TI - Regulation of the pluripotency marker Rex-1 by Nanog and Sox2. AB - Rex-1 (Zfp-42) is a known marker for undifferentiated embryonic stem cells and teratocarcinoma cells. However, the mechanism by which Rex-1 is regulated in pluripotent cells remains unresolved. Here we report that Nanog, an Nk-2 homeodomain protein known for its role in maintaining stem cell pluripotency, is a transcription activator for the Rex-1 promoter. Knockdown of Nanog in embryonic stem cells resulted in a reduction of Rex-1 expression, whereas forced expression of Nanog in P19 stimulated Rex-1 expression. Employing a Rex-1 reporter, we demonstrate that Nanog transactivates Rex-1 directly. Serial deletion studies mapped the Nanog-responsive element between -187 and -286 of the Rex-1 promoter. Although Oct-3/4 and Sox2 can both transactivate Rex-1 promoter, only Sox2 cooperates with Nanog in up-regulating Rex-1. Furthermore, we demonstrate that the C terminus of Nanog is responsible for transactivating the Rex-1 promoter, a function that can be substituted for by a viral transactivator Vp16 efficiently in NIH3T3 cells but less so in P19 cells. Taking these findings together, we conclude that Rex-1 is a direct target of Nanog, which is augmented by Sox2 and Oct-3/4. PMID- 16714767 TI - Comparison of two fluid-management strategies in acute lung injury. AB - BACKGROUND: Optimal fluid management in patients with acute lung injury is unknown. Diuresis or fluid restriction may improve lung function but could jeopardize extrapulmonary-organ perfusion. METHODS: In a randomized study, we compared a conservative and a liberal strategy of fluid management using explicit protocols applied for seven days in 1000 patients with acute lung injury. The primary end point was death at 60 days. Secondary end points included the number of ventilator-free days and organ-failure-free days and measures of lung physiology. RESULTS: The rate of death at 60 days was 25.5 percent in the conservative-strategy group and 28.4 percent in the liberal-strategy group (P=0.30; 95 percent confidence interval for the difference, -2.6 to 8.4 percent). The mean (+/-SE) cumulative fluid balance during the first seven days was -136+/ 491 ml in the conservative-strategy group and 6992+/-502 ml in the liberal strategy group (P<0.001). As compared with the liberal strategy, the conservative strategy improved the oxygenation index ([mean airway pressure x the ratio of the fraction of inspired oxygen to the partial pressure of arterial oxygen]x100) and the lung injury score and increased the number of ventilator-free days (14.6+/ 0.5 vs. 12.1+/-0.5, P<0.001) and days not spent in the intensive care unit (13.4+/-0.4 vs. 11.2+/-0.4, P<0.001) during the first 28 days but did not increase the incidence or prevalence of shock during the study or the use of dialysis during the first 60 days (10 percent vs. 14 percent, P=0.06). CONCLUSIONS: Although there was no significant difference in the primary outcome of 60-day mortality, the conservative strategy of fluid management improved lung function and shortened the duration of mechanical ventilation and intensive care without increasing nonpulmonary-organ failures. These results support the use of a conservative strategy of fluid management in patients with acute lung injury. (ClinicalTrials.gov number, NCT00281268 [ClinicalTrials.gov].). PMID- 16714768 TI - Pulmonary-artery versus central venous catheter to guide treatment of acute lung injury. AB - BACKGROUND: The balance between the benefits and the risks of pulmonary-artery catheters (PACs) has not been established. METHODS: We evaluated the relationship of benefits and risks of PACs in 1000 patients with established acute lung injury in a randomized trial comparing hemodynamic management guided by a PAC with hemodynamic management guided by a central venous catheter (CVC) using an explicit management protocol. Mortality during the first 60 days before discharge home was the primary outcome. RESULTS: The groups had similar baseline characteristics. The rates of death during the first 60 days before discharge home were similar in the PAC and CVC groups (27.4 percent and 26.3 percent, respectively; P=0.69; absolute difference, 1.1 percent; 95 percent confidence interval, -4.4 to 6.6 percent), as were the mean (+/-SE) numbers of both ventilator-free days (13.2+/-0.5 and 13.5+/-0.5; P=0.58) and days not spent in the intensive care unit (12.0+/-0.4 and 12.5+/-0.5; P=0.40) to day 28. PAC-guided therapy did not improve these measures for patients in shock at the time of enrollment. There were no significant differences between groups in lung or kidney function, rates of hypotension, ventilator settings, or use of dialysis or vasopressors. Approximately 90 percent of protocol instructions were followed in both groups, with a 1 percent rate of crossover from CVC- to PAC-guided therapy. Fluid balance was similar in the two groups, as was the proportion of instructions given for fluid and diuretics. Dobutamine use was uncommon. The PAC group had approximately twice as many catheter-related complications (predominantly arrhythmias). CONCLUSIONS: PAC-guided therapy did not improve survival or organ function but was associated with more complications than CVC guided therapy. These results, when considered with those of previous studies, suggest that the PAC should not be routinely used for the management of acute lung injury. (ClinicalTrials.gov number, NCT00281268.). PMID- 16714769 TI - Fluid-management strategies in acute lung injury--liberal, conservative, or both? PMID- 16714770 TI - Pulmonary-artery catheters--peace at last? PMID- 16714771 TI - Is the ApoE polymorphism associated with dilated cardiomyopathy? AB - Apolipoprotein E (ApoE) is 34 kDa protein involved in the modulation of cholesterol transport and homeostasis. Polymorphism of the ApoE gene has been implicated in many chronic cardiovascular and neuronal diseases. ApoE epsilon4 allele has been reported to be associated with increased risk of cardiovascular diseases such as myocardial infarction, hypertension, coronary heart disease, etc. Fifty patients with the end-stage dilated cardiomyopathy (DCM) and advanced congestive heart failure were examined in our study. For evaluation of ApoE polymorphism, novel approach of fast screening of ApoE gene polymorphism by combination of PCR and blotting (CVD StripAssay) was used. Individual genotypes were correlated with basic cardiologic clinical parameters. The reported frequency of this allele in Caucasian population is 14.7 %. Our results showed that in patients with DCM frequency of the ApoE epsilon4 allele is 40 %. Frequency of the genotype epsilon2/4 was 58 % and epsilon3/4 was 22 %. Comparison with control Caucasian groups monitored by others clearly revealed that frequency of epsilon4 alelle is increased in patients with advanced stages of DCM. This observation suggests association of ApoE polymorphism with severe form of DCM. Physiological consequences of this observation remain to be clarified. PMID- 16714772 TI - Time dependent effects of dexamethasone on serum insulin level and insulin receptors in rat liver and erythrocytes. AB - The effects of glucocorticoid excess on regulation of insulin receptors were investigated in dexamethasone-treated rats. Glucocorticoid excess was produced by administration of dexamethasone (0.5 mg/100 g b.w.) 30 min, 4, 12, 18, 24, 42 or 70 h before experiments. This treatment caused time-dependent changes of glucose and insulin concentration in blood, as well as in amounts of specific insulin binding and insulin receptors of liver cells and erythrocytes. The time intervals in which dexamethasone produced the increase in insulin concentration were accompanied with decrease in insulin binding to receptors in membranes of liver cells, while significant changes in insulin binding to receptors of erythrocytes were not observed under the same experimental conditions. The effect is maximal 18 and 42 h after dexamethasone treatment that increase insulin blood level by about 85% and 60%, respectively. Receptor analysis revealed that changes in specific binding of insulin could be due to significant changes in amount of binding sites on cell surface rather than to mild alteration in receptor affinity. These findings suggest that besides the changes in insulin level, the alterations in insulin receptor number and affinity may play a major role in the states of altered insulin sensitivity which accompany glucocorticoid excess. PMID- 16714773 TI - Changes in the expression and/or activation of regulatory proteins in rat hearts adapted to chronic hypoxia. AB - Chronic intermittent high altitude (IHA) hypoxia results in long-term adaptation protecting the heart against acute ischemia/reperfusion injury; however, molecular mechanisms of this phenomenon are not completely elucidated so far. The present study was aimed at investigation of a modulating effect of IHA hypoxia on the expression and/or activation of selected regulatory proteins, with particular emphasis on differential responses in the right ventricle (RV) and left ventricle (LV). Adult male Wistar rats were exposed to IHA hypoxia of 7000 m simulated in a hypobaric chamber (8 h/day, 25 exposures), and protein contents and activities in myocardial fractions were determined by Western blot analysis. In markedly hypertrophic RV of hypoxic rats, gelatinolytic activity of MMP-2 and protein levels of carbonic anhydrase IX (a marker of hypoxia) were significantly enhanced. Study of mitogen-activated protein kinases (MAPKs) revealed no differences in the contents of total p38-MAPK in both ventricles between the IHA and normoxic control rats, whereas activation of p38-MAPK was decreased in the RV and moderately increased in the LV of IHA rats as compared to controls. Extracellular signal regulated kinase-2 (ERK-2) was partially up-regulated in the RV of IHA rats, and, in addition, expression of acidic fibroblast growth factor (aFGF), a potential activator of ERK cascade, was also significantly increased. In contrast, expression of ERKs in the LV as well as their activities in both ventricles, were not affected by IHA hypoxia. Differential effects of IHA hypoxia on c-Jun-N-terminal protein kinases (JNKs) in the RV and LV were also observed. As compared with the controls, total content of JNKs was increased in the RV of the IHA rats, while expression of JNKs in the LV was down-regulated. IHA hypoxia changed neither total levels of Akt kinase in both RV and LV, nor Akt kinase activity in the RV. However, increased levels of activated phospho-Akt kinase were found in the LV of IHA rats. The results demonstrate that adaptation of rat hearts to chronic IHA hypoxia is associated with disctinct changes in the levels and/or activation of several regulatory proteins in two ventricles. The latter could be attributed to both myocardial remodeling and cardioprotection induced by chronic hypoxia. PMID- 16714774 TI - Blockade of AT1 receptors by losartan did not affect renin gene expression in kidney medulla. AB - This study was designed to determine particular changes in the renin gene expression and activity in renal cortex and medulla after AT(1) receptor blockade. It was found that two-week-treatment with AT(1) blocker losartan induced an increase in tissue renin activity in both parts of kidney causing subsequent elevation of plasma renin activity. Renin mRNA in losartan-treated rats was increased only in cortex, suggesting cortex origin of elevated renin activity in medulla. Medullary renin mRNA indicated local synthesis of renin within the whole kidney and supported the idea of the presence of tissue renin angiotensin system. Our results show that gene expression of renin in kidney medulla is insensitive to AT(1) receptor blockade and this points out that the regulation of kidney renin-angiotensin system probably differs from that in cortex. PMID- 16714775 TI - Mechanistic equations for membrane transport of multicomponent solutions. AB - In the present article, mechanistic equations for membrane transport of N + 1 component solutions have been derived. The major specific investigation result is the introduction - for ternary solutions - of two diffusion coefficients omega(d1) and omega(d2) for solutes, as well as two cross coefficients omega(d12) and omega(d21) for these solutes. The latter parameters may be treated as coefficients of interdiffusion. The expansion of the description of substance transport to include the N + 1-component solutions does not formulate any additional physical phenomena other than those which are formulated by the transport equations for three-component solutions. PMID- 16714776 TI - Effect of valproic acid and antiapoptotic cytokines on differentiation and apoptosis induction of human leukemia cells. AB - This work compares effect of histondeacetylase inhibitor, valproic acid (VA), on proliferation, differentiation and apoptosis induction in two human leukemic cell lines: HL-60 (human promyleocytic leukemia, p53 negative) and MOLT-4 (human T lymphocyte leukemia, p53 wild type). Incubation with VA caused decrease in percentage of cells in S phase of cell cycle. The decrease was more intensive in HL-60 cells, where the cells in S phase were absent 6 days after the beginning of incubation with VA (4 mmol/l). 3-day-long incubation of HL-60 cells with 4 mmol/l VA caused differentiation of these cells, marked by increase in CD11b and co stimulatory/adhesion molecule CD86, and induction of a significant apoptosis. Annexin V positive cells lost the CD11b antigen. 3-day-long incubation of MOLT-4 cells with VA (1-2 mmol/l) inhibited proliferation and decreased percentage of cells in S phase of the cell cycle. 90% of MOLT-4 cells are CD7 positive. This CD7 positivity is not changed during apoptosis induction (detected as Annexin V positivity). On the other hand, CD4 marker expression decreases after incubation with 1-2 mmol/l VA, but during apoptosis induction by 4 mmol/l VA, most of the apoptotic Annexin V positive cells were also CD4 positive. Using a clonogenic survival assay EC(50) for 3-day-long incubation with VA was determined. For HL-60 cells, the established EC(50) was 1.84 mmol/l, for MOLT-4 cells it was 1.76 mmol/l. Ability of VA to induce differentiation in HL-60 cells thus does not affect final cell killing. However, the elimination of the cells was considerably affected by presence of hematopoietic growth factors. 14-day-long incubation of HL-60 cells with VA in conditioned medium (source of IL-3, SCF, G-CSF) caused increase in EC(50) to 4 mmol/l, while in MOLT-4 cells (cultivation without conditioned medium), the EC(50) decreased to 0.63 mmol/l. PMID- 16714777 TI - Effect of pentoxifylline on endothelaemia and hypothalamic-pituitary adrenocortical axis activation in female rats under stress exposure. AB - Endothelial dysfunction may belong to negative consequences of stress exposure accompanied by activation of several stress systems including the hypothalamic pituitary-adrenocortical (HPA) axis. The present experiments were aimed at testing the hypotheses that i) immobilization (IMO) stress results in sustained increase in endothelaemia for 24 h and that ii) pentoxifylline, a drug with endothelium protective properties, attenuates the rise in endothelaemia and HPA axis activation in female rats as shown previously in males. Circulating endothelial cells increased immediately after the IMO for 2 h, returned back to control levels at 12 h and increased again at 24 h. Stress-induced rise in adrenocorticotropic hormone (ACTH) and corticosterone levels was particularly high immediately after the IMO. Pretreatment with pentoxifylline (20 mg/kg subcutaneously for 7 days) attenuated the rise in endothelaemia and adrenal corticosterone measured at 24 h following IMO. Plasma levels of ACTH and proopiomelanocortin gene expression in the anterior pituitary were not affected by pentoxifylline treatment. The present results indicate that IMO stress in female rats induces a biphasic rise in endothelaemia early at the time of stress exposure and than 24 h thereafter. Based on these data and our previous study we can conclude that intensive stress has a negative influence on endothelial cells in both sexes and no gender differences seem to be present in the protective action of pentoxifylline. PMID- 16714778 TI - Inhibition of rat brain ecto-atpase activity by various drugs. AB - The in vitro effect of digoxin, verapamil, propranolol, carbamazepine, diazepam and promethazine were investigated on the ecto-ATPase activity of synaptosomal plasma membranes from the rat brain. ATP hydrolyzing activities of the enzyme were not affected by digoxin while the use of all other drugs resulted in significant and dose-dependent ihibition in ATP hydrolysis. According to values of IC(50) and K(iapp), the order of inhibitory potency of the drugs applied was: diazepam > promethazine > verapamil > propranolol >> carbamazepine. Kinetic analysis of the nature of the ATPase inhibition revealed that it resulted from a direct action of drugs on the enzyme protein. The aim of the present study was to determine the potential neuromodulatory side effects of the drugs investigated. The results achieved indicated that all investigated drugs, except digoxin, may modulate neuronal activities via the purinergic receptors P2 by increasing extracellular concentrations of ATP as a consequence of inhibition of the ecto ATPase activity. Our findings indicate that it may be useful to take into consideration the possible side effects of the investigated drugs, when they are used in treatment of different pathologies, particularly in the treatment of epilepsy by carbamazepine and diazepam. PMID- 16714779 TI - Adaptive Silicon Monochromators for High-Power Insertion Devices. Tests at CHESS, ESRF and HASYLAB. AB - X-ray wigglers which produce tens of kilowatts of photon power within the white beam will soon become available at third-generation sources of synchrotron radiation. Insertion devices that produce several kilowatts already exist and we have used those at CHESS, ESRF and HASYLAB to test adaptive 111 silicon water-jet cooled monochromators at up to 2 kW total incident beam power. This development from earlier work at the Brookhaven National Synchrotron Light Source (NSLS) uses the pressure in the water coolant to provide active compensation of the strain field in the thermal footprint, nulling its effect to within residual variations in Bragg angle of only a few arc s. The design is robust, vacuum compatible and uses no moving mechanical parts. PMID- 16714780 TI - Fixed-exit monochromators for high-energy synchrotron radiation. AB - Current developments in X-ray optics for synchrotron radiation beamlines are briefly reviewed. Reference is made to recent work on adaptive mirrors, cryogenic cooling of monochromators, use of thin diamond crystals, and active correction of the crystal shape for distortions caused by beam heating. The use of bent Si crystals as monochromators is discussed in detail. At high energies Si crystals become transparent to X-rays allowing new monochromator constructions. Bending of the crystal increases the energy bandpass and allows focusing. Different combinations of bent crystals that provide a fixed exit beam are discussed. These include vertically diffracting meridionally focusing Laue-Laue crystals, a Laue crystal combined with a sagittally focusing Bragg crystal, and a Laue-Bragg pair of crystals which provides meridional focusing at two stages. PMID- 16714781 TI - Imaging plates for use with synchrotron radiation. AB - This review summarizes the principles and performance characteristics of the imaging-plate (IP) X-ray area detector, which is based on the photo-stimulable phosphor BaF(Br,I):Eu(2+), together with some of its applications at the Photon Factory. The photo-stimulable phosphor can temporarily store an X-ray image. The stored image is read out by measuring the intensity of luminescence, which is stimulated by an He-Ne laser beam scanning the phosphor surface. The IP has a spatial resolution of 170 mum (FWHM) with a pixel size of 100 x 100 mum and area sizes ranging from 127 x 127 to 201 x 400mm(2). The dynamic range is over l:10(5). The detective quantum efficiency, which is a function of exposure level, is more than 80% for 8-20 keV X-rays at medium exposure levels. The background noise level is equivalent to less than 3 X-ray photons/pixel of 8 keV. The precision in intensity measurement is 0.5-1% at best. These performance characteristics of the IP depend largely on the performance of the IP readout system. Two applications of the IP to time-resolved measurements are discussed: one is based on a cinema method which achieves a 0.3 s time resolution for 40 exposures of size 127 x 127 mm(2), and the other uses the IP as a linear detector of length 200 mm with a 23 mus time resolution for a time period of 46 ms, based on the streak-camera method. PMID- 16714782 TI - Image-plate synchrotron laue data collection and subsequent structural analysis of a small test crystal of a nickel-containing aluminophosphate. AB - Image plates have advantages over photographic films, which include wider dynamic range, higher detector quantum efficiency, reduced exposure time and large size. In this study, an on-line image-plate system has been used to record crystallographic data from a small crystal. In particular, synchrotron Laue data were recorded with lambda(min) = 0.455, lambda(max) = 1.180 A, in 20 images 10 degrees apart and with an exposure time of 0.3 s each from a crystal (0.02 x 0.05 x 0.25 mm) of a nickel-containing aluminophosphate, NiAPO. The Laue data were analyzed with the Daresbury Laue software, including the application of an absorption correction. The structure was solved by a combination of the Patterson method and successive difference Fourier calculations using SHELXS86 and SHELXL93; the final R value for 1934 unique reflections (all data) and 310 parameters was 7.90%. The structure agrees with that determined by monochromatic diffractometry using the same crystal and reported by Helliwell, Gallois, Kariuki, Kaucic & Helliwell [Acta Cryst. (1993), B49, 420-428] with an r.m.s. deviation of 0.03 A. Hence, this study shows the image-plate device to be very effective for synchrotron data collection and subsequent structure analysis from small crystals, i.e. 0.02 x 0.05 x 0.25 mm, in chemical crystallography as well as providing further confirmation of the practicability of Laue data in structure solution and refinement. PMID- 16714783 TI - Evaluation of Reflection Intensities for the Components of Multiple Laue Diffraction Spots. III. Using a Real-Space Density Modification Method. AB - In the Laue diffraction pattern, 10--20% of the spots result from the exact superposition of two or more reflections that are ;harmonics', e.g. hkl; 2h 2k 2l, ...; a high proportion of these are low-resolution reflections. For the solution of large or difficult structure problems, the intensities of the remaining 80-90% of the reflections, measurable as singles, may not be sufficient and thus the evaluation of the intensities of the components of the multiple spots is important. A new procedure, DECONV, for this deconvolution using real space density modifications on the Patterson map is given. This development is based on a procedure in reciprocal space related to direct methods [Hao, Campbell, Harding & Helliwell (1993). Acta Cryst. A49, 528-531]. A Patterson map is calculated using single reflections and modified by removing negative densities and squaring with an appropriate adjustment at the origin peak. The procedure can be repeated until convergence is reached. It has been tested with Laue diffraction data from 4Zn insulin and cytochrome c peroxidase (CCP). 304 and 1134 reflection intensities were evaluated from multiple spots of insulin and CCP, respectively; the mean fractional differences (on F), showing the agreement with high-quality monochromatic data, were 0.27 and 0.21, respectively. PMID- 16714784 TI - Polarized Dispersion for X-rays Scattered by an Aromatic Bromide. AB - Second-rank tensors describing how the real and imaginary dispersion terms f' and f'' depend on photon polarization for Br atoms substituted on a benzene ring have been measured at six energies near the Br K absorption edge in diffraction experiments with synchrotron radiation and crystals of homocubanecarboxylic acid p-bromoanilide. Resonance with a sigma antibonding orbital causes f' to change with polarization as much as 4.3 and f''' as much as 7.5. The tensors for two independent Br atoms are equal, uniaxial and aligned with the respective Br-C bond. Absorption spectra show the average effect of two tensors with different orientations. Atomic coordinates from a new refinement of the structure are listed. PMID- 16714785 TI - The Multiwavelength Anomalous Solvent Contrast (MASC) Method in Macromolecular Crystallography. AB - The wavelength dependence of anomalous scattering of X-rays, due to atoms randomly dispersed in the solvent phase of a macromolecular crystal, is a way of producing solvent-density contrast variation with perfect isomorphism. The largest contrast variations are obtained by tuning the X-ray wavelength near an absorption edge of the anomalous-scattering species. In this method, which we call MASC, the anomalous partial structure is an extended uniform electron density, in contrast to the few punctual ordered scatterers in the multiwavelength anomalous-dispersion (MAD) method. MASC is, in principle, applicable to the determination of the molecular envelope and of low-resolution structure-factor phases. Structure factors (lambda)F(+/-h) leads to a set of equations which can be solved to give |G(h)| and |(0)F(h)|, the modulus of the envelope and of the total ;normal' structure factors, respectively, and Deltavarphi = (varphi(0)(F)-varphi(G)). The moduli {|G|} behave like structure factor amplitudes from small-molecule crystals, and the estimation of their phases can be carried out by statistical direct methods. Then, the phase of (0)F(h) and finally the conventional (e.g. in vacuum) protein structure factor F(p)(h) can be determined. As in the MAD method, the strength of MASC signals can be quantified by Bijvoet and dispensive ratios, for which practical expressions are derived in the case of zero contrast. The behaviour of these ratios at increasing resolution is discussed, using approximations for |G(h)| and |Delta(h)| , respectively, derived from Porod's law and assuming a random distribution of atoms in the solvent excluding volume. Expected values of anomalous ratios are calculated for a hypothetical MASC experiment based on the known three-dimensional structure of kallikrein A, using a solvent with 3.5 M ammonium selenate to ensure zero contrast, and wavelength tuning near the Se K absorption edge. The main steps of a MASC experiment are discussed in the context of a MAD-like data collection optimized for accurate measurements of intensities of anomalous pairs at low resolution. Finally, the results of preliminary experiments on two protein crystals are reported. The first, a partial single wavelength data collection, used anomalous scattering of selenium at the K edge and gave anomalous ratios with the expected behaviour. The second one, at three wavelengths, used anomalous scattering of ytterbium at the L(III) edge. In this case, effects from solvent as well as from ordered lanthanide ions were demonstrated. PMID- 16714786 TI - Rapid mapping of texture in polycrystalline materials using an imaging plate on a synchrotron radiation source. AB - Taking advantage of the high brilliance of synchrotron radiation, a system was developed for rapid mapping of the orientation distribution of crystal grains (texture) in polycrystalline materials using an imaging plate. A monochromatized beam is incident on the sample, which is rotated using the omega-axis mechanism of an X-ray diffractometer so that the surface of the sphere of poles of the selected reflection is scanned by the Ewald sphere. Simultaneously, the imaging plate is translated vertically with a velocity that is synchronized with that of the sample rotation. It is possible to record pole figures over an extended angular range within a short period of time, typically of the order of minutes. The method has been applied to the observation of a time change in the orientation distribution of metal sheets at elevated temperatures. PMID- 16714787 TI - Interpretation of EXAFS Data. AB - An outline of the theory of X-ray absorption as applied to X-ray absorption spectroscopy is given, concentrating on EXAFS, showing how structural parameters are contained within the theory. A full description of the scattering problem and of inelastic effects is also included, and a description is given of how structural information may be extracted from experimental data. The emphasis is on curve-fitting methods, including those techniques which allow the inclusion of external constraints. PMID- 16714788 TI - Quick Fluorescence-EXAFS: an Improved Method for Collection of Conventional XAFS Data, an Improved Method for Collection of Conventional XAFS Data and for Studying Reaction Intermediates in Dilute Systems. AB - The quick EXAFS (QuEXAFS) technique provides an alternative way of recording X ray absorption fine-structure (XAFS) data where the scan time is reduced by moving the monochromator at a constant angular speed and recording the data ;on the fly'. Results are presented to show that the use of fluorescence detection with QuEXAFS is eminently suitable for studying reactions in dilute systems such as metalloproteins at a sub-minute time scale. In addition, we show that the fluorescence-QuEXAFS technique can reduce the overall time for normal data collection by some 50% compared with conventional step-by-step scanning EXAFS using the same optical system, thus reducing the total X-ray exposures of the samples. The use of X-rays for studying in situ redox reactions is demonstrated. PMID- 16714790 TI - New commercial products. PMID- 16714791 TI - A Multiwavelength Monochromator for the X3A1 Fixed-Angle Station at the National Synchrotron Light Source. AB - A new revolving monochromator intercepting 2 mrad of the radiation fan at the State University of New York (SUNY) X3 beamline at the National Synchrotron Light Source (NSLS) is described. The design allows for easy interchange between four different monochromator crystals. The photons are reflected in the horizontal plane to provide a monochromatic beam at a fixed-angle diffraction station. The triangular crystals can be bent for horizontal focusing. PMID- 16714792 TI - Time differences between friedel reflections: accuracy of crystal setting and requirements on beam stability. AB - In synchrotron radiation data collections where the wavelength is carefully set to optimize f''-derived crystallography intensity differences (Friedel pairs), careful alignment of the crystal is useful to minimize the time differences of stimulation of the reflections in the pairs. This paper quantifies these time differences as a function of crystal misorientation, with typical parameters, using the angular velocity of the crystal. Likewise, the time spent in the diffraction condition is also calculated via the angular reflecting range for a common synchrotron beam geometry. These times offer the user direct insight into the time-dependent aspects of the diffraction measurements. This therefore allows the optimum conditions to be set up so as to extract as accurate anomalous differences as possible in the context of synchrotron radiation beam stability and lifetime. PMID- 16714794 TI - Real-time simultaneous wide- and small-angle fibre diffraction. AB - A combination of two independent imaging area-detector systems controlled by a single data-acquisition system, provides a powerful system for X-ray diffraction studies of time-resolved phenomena over a wide q range, in samples with intrinsic or induced structural orientation. With this system we have observed a transient, tensile-stress-induced, orthorhombic-to-monoclinic transition in high-density polyethylene. PMID- 16714795 TI - 2p X-ray Absorption Spectroscopy in the Earth Sciences. AB - A complete knowledge of 3d transition-metal valencies, site occupancies and site symmetries is essential for a full understanding of mineral/melt energetics and behaviour. Over the last few years, significant advances in both instrumentation and theory associated with synchrotron radiation sources and experiments have enabled the development of 2p X-ray absorption spectroscopy as a sensitive, element-specific site and valency probe. The potential of this technique in the Earth sciences is discussed in this paper with examples reflecting the variety of problems set by 3d transition metals in natural systems. PMID- 16714793 TI - Anomalous Dispersion with Edges in the Soft X-ray Region: First Results of Diffraction from Single Crystals of Ribosomes Near the K-Absorption Edge of Phosphorus. AB - X-ray diffraction from a single crystal of the small ribosomal subunit of Thermus thermophilus was measured at five wavelengths near the K-absorption edge of phosphorus. The intensity of the low-order diffraction peaks varies strongly with the wavelength. It is influenced by the dispersion of contrast due to the P atoms of the ribosomal ribonucleic acid and by that of absorption, in comparable amounts. PMID- 16714796 TI - Surface-Sensitive XAFS in the Hard X-ray Region with Sub-Monolayer Sensitivity. AB - Surface-sensitive X-ray absorption fine structure (XAFS) with sub-monolayer sensitivity based on grazing-incidence fluorescence detection is reported. The efficiency of fluorescence detection increased by more than two orders of magnitude by combining a multipole wiggler with a multi-element Si(Li) solid state detector. The capability of the present technique for structural studies of surfaces and buried interfaces in the hard X-ray region was demonstrated by As K edge XAFS studies of the InP(001) surface exposed to AsH(3) flow. The results indicated that ~0.1 monolayer As atoms are incorporated into the surface replacing the P atoms. PMID- 16714797 TI - Application of photoelectron spectromicroscopy to a systematic study of toxic and natural elements in neurons. AB - A systematic photoelectron spectromicroscopy study is presented of the spatial distribution of a toxic element, aluminium, iron or chromium, in neuron cultures, after exposure to a solution of the element. The study was performed by the X-ray secondary-emission microscopy (XSEM) version of photoelectron Spectromicroscopy. The distribution of the elements was investigated with two complementary approaches: digital subtraction imaging and individual X-ray absorption spectra from microscopic areas. The results coherently indicate different localization patterns for different elements, and, in particular, extreme localization of aluminium to a few rare cells identifiable as Purkinje neurons. In the case of iron-exposed specimens, the distribution analysis was extended to naturally present phosphorus, and used to estimate the XSEM sensitivity. PMID- 16714799 TI - A Source Design Strategy Providing 5 eV-100 keV Photons. Erratum. AB - An error in the paper by Suller [J. Synchrotron Rad. (1994). 1, 5-11] is corrected. The units for the ordinate in the caption for Fig. 7 should be photons s(-1) (0.1% bandwidth)(-1) mm(-2) mrad(-2) and photons s(-1) (0.1% bandwidth)( 1), respectively. PMID- 16714798 TI - Search for new scintillators by studying the fluorescence properties of powdered compounds with synchrotron radiation. AB - This paper describes a method for investigating the properties of powdered compounds using synchrotron X-radiation. The fluorescence decay times of bismuth germanate, BaF(2), CeF(3) and CsI, as well as the light yields of several samples of gadolinium silicate, prepared in different ways, have been measured. PMID- 16714800 TI - Nominations for the ewald prize. PMID- 16714801 TI - New commercial products. PMID- 16714802 TI - Adaptive silicon monochromators for high-power wigglers; design, finite-element analysis and laboratory tests. AB - Multipole wigglers in storage rings already produce X-ray power in the range up to a few kilowatts and planned devices at third-generation facilities promise up to 30 kW. Although the power density at the monochromator position is an order of magnitude lower than that from undulators, the thermal strain field in the beam footprint can still cause severe loss of performance in X-ray optical systems. For an optimized adaptive design, the results of finite-element analysis are compared with double-crystal rocking curves obtained with a laboratory X-ray source and, in a second paper [Quintana, Hart, Bilderback, Henderson, Richter, Setterson, White, Hausermann, Krumrey & Schulte-Schrepping (1995). J. Synchotron Rad. 2, 1-5], successful tests at wiggler sources at CHESS and ESRF and in an undulator source at HASYLAB are reported. PMID- 16714803 TI - A Double Multilayer Monochromator at an ESRF Undulator for Microbeam Experiments. AB - A water-cooled double W/Si-multilayer monochromator has been operated at an ESRF low-beta undulator beam. For a fixed distance of the two multilayers the first order Bragg reflection was at ~8 keV. The peak power density of the beam at the exit of the multilayers was ~1 W mm(-2) and the flux density of the first order after a 10 mum collimator was 4 x 10(5) photons s(-1) mum(-2) mA(-1.) The performance of the beam in microbeam diffraction has been tested on a 20 mum W wire. The observed pseudo-Laue pattern is discussed with respect to the multilayer spectrum. PMID- 16714804 TI - Observation of Interference Effects at the Focus of an X-ray Lens. AB - During an experiment on synchrotron radiation focusing with a capillary lens, an interference structure was observed at the focal spot of the lens, despite the fact that the lens capillary diameter is about a million times greater than the wavelength of the X-ray photons (600 mum and 8 A, respectively). The width of the central peak is close to the capillary diameter. At the same time the synchrotron radiation concentration increased by more than one hundred times. Analysis shows that the capillary lens acts in many respects as a macroscopic crystal. The observed effect is accounted for by wave theory. This phenomenon may have important practical effects in many fields. PMID- 16714805 TI - The Perfection of Protein Crystals Probed by Direct Recording of Bragg Reflection Profiles with a Quasi-Planar X-ray Wave. AB - Profiles of Bragg reflections from earth-grown crystals of lysozyme from hen egg white and collagenase from Hypoderma lineatum were directly recorded with a quasi planar X-ray wave. One crystal of each protein was chosen for a detailed investigation. Each sample is shown to consist of only a few (three and two, respectively) highly ordered domains, misoriented with respect to each other by a few arc s. The smallest rocking widths were observed for the large domain of the collagenase sample (FWHM corrected for instrumental broadening: 0.0016 degrees for a strong reflection at 3 A resolution). With appropriate improvements, this method might become a quantitative tool for characterizing the perfection of crystals from biological macromolecules. PMID- 16714806 TI - Long Horizontal Parallel Slits with 0.03 degrees Angular Resolution for Powder Diffraction Using Synchrotron Radiation. AB - Long horizontal parallel slits with angular apertures of 0.032 and 0.065 degrees were constructed for powder diffraction experiments with synchrotron radiation. They have been tested at the BL-4B experimental station at the Photon Factory by using a monochromated beam with a wavelength of 1.528 A. The horizontal parallel slits with the smaller aperture gave a full-width at half-maximum of 0.030 (1) degrees for the (200) reflection from CeO(2) and an intensity about one order of magnitude higher than that obtained with a receiving slit in the same angular resolution, demonstrating the finest horizontal parallel slits developed so far. The misalignment of the horizontal parallel slits does not affect the intensity whilst it shifts the Bragg-peak positions systematically. PMID- 16714809 TI - Notes and news. PMID- 16714807 TI - A Primer in Synchrotron Radiation: Everything You Wanted to Know about SEX (Synchrotron Emission of X-rays) but Were Afraid to Ask. AB - The basic properties of synchrotron radiation are derived with simple approaches, emphasizing phenomena rather than mathematical details. PMID- 16714810 TI - Perfect Crystals in the Asymmetric Bragg Geometry as Optical Elements for Coherent X-ray Beams. AB - Perfect crystals in the asymmetric Bragg geometry are evaluated as optical elements for manipulating coherent X-ray beams. Such optics can be used to modify the transverse coherence length of a synchrotron X-ray beam, with the intention of increasing the usable coherent flux. The wavelength range, angular divergence and flux of X-rays passing through a pinhole aperture are examined in detail, as functions of source and pinhole size, crystal-to-pinhole separation and the asymmetry factor. In developing this analysis, the behavior of asymmetrically cut crystals is explained in reciprocal space, with reference to the crystal truncation rod associated with the reflection. The results show that, for synchrotron beams that are collimated to a small fraction of the incident Darwin width, the wavelength range accepted by the crystal is typically dispersed into an angular spread in the exit beam. This chromatic aberration greatly reduces the transverse coherence length in a manner that does not conserve the coherent flux. The calculations are in agreement with measurements of the divergence and flux through a micrometer-sized pinhole using a synchrotron wiggler X-ray source. PMID- 16714811 TI - Curved Crystal Transmission Optics for Energy-Dispersive X-ray Absorption Spectroscopy. AB - The development of a curved crystal monochromator of the Laue type for energy dispersive X-ray absorption spectroscopy is presented. The quality of the X-ray absorption spectra at high photon energies is compared with spectra measured with silicon crystals in the more frequently used Bragg geometry. In the Bragg case, an asymmetric broadening of the reflectivity profile leads to strong distortions of the near-edge fine structure and to a reduction in spectral resolution. The reflectivity profiles of fiat and curved crystals for Laue and Bragg geometry have been calculated using dynamical theory and are compared with experimental data. The new optics have been used for in situ time-resolved X-ray absorption spectroscopy. An example of the application of the technique for the characterization of a Pd catalyst is given. The X-ray absorption fine structure at the Pd K-edge has been measured during the activation and during the heterogeneous catalytic oxidation of carbon monoxide. PMID- 16714812 TI - The Use of Imaging Plates in Electron-Density Mapping with Synchrotron X-rays. AB - Imaging-plate synchrotron data have been applied in the charge-density analyses of sodium nitroprusside and hexaamminechromium(III) hexacyanochromate(lll), collected at 100 and 50 K, respectively, and photons of wavelengths 0.656 and 0.394 A at the SUNY X3 beamline at NSLS. The electron-density maps show good agreement between chemically equivalent sections, while the multipole aspherical atom refinements lead to chemically reasonable population parameters, with trends reproduced in the available theoretical calculations. The results indicate that the time required for charge-density mapping with diffraction data can be greatly reduced by the application of the new technology. PMID- 16714813 TI - Application of synchrotron radiation laue diffraction to small single crystals of a mineral - structure determination and identification. AB - The potential of synchrotron radiation Lane diffraction Patterns for structure determination and for identification of small single crystallites is illustrated. Diffraction Patterns were recorded for several crystallites, 4f and transition-metal 3p --> 3d thresholds have been carried out using a variety of complex cuprates and nickelates on stations 6.1 (grazing-incidence monochromator) and 6.2 (toroidal-grating monochromator) at the SRS CLRC Daresbury Laboratory. The systems studied are Nd(2)Ni(1 - x)Cu(x)O(4), La(2 - x)Sr(x)Ni(1- y)Fe(y)O(4 + delta) and Bi(2)Sr(2)Ca(1 - x)Y(x)Cu(2)O(8 + delta). A combination of EDC and constant-initial-state data is used to examine the 4f and 3d contributions to the valence-band density of states and their binding-energy positions relative to the Fermi energy. This allows the study of the valence states of the transition-metal ions and their modulation on doping. For La(2 - x)Sr(x)Ni(1 - y)Fe(y)O(4 + delta), this approach is used to infer a valence state of >/= 3.0 for Fe. In the case of Bi(2)Sr(2)Ca(1 - x)Y(x)Cu(2)O(8 + delta), the effect of Cu valence modulation on the 3p resonance is observed as x is varied. This is discussed in the light of controversy surrounding shifts in core-level photoemission with doping for this system. PMID- 16714829 TI - A Perspective: JSR so far. PMID- 16714831 TI - Compensation of Aberration of Inclined X-ray Monochromators. AB - Inclined double-crystal monochromator for synchrotron radiation suffer from aberration that is connected with the horizontal divergence of the synchrotron radiation beam. Two different methods are proposed to compensate for this aberration. The first method introduces slightly different angles of inclination for the first and the second monochromator crystals. The condition for the difference of the angles of inclination is Bragg angle dependent and also depends on the projection of the distance of both crystals onto the direction of the normal to the diffracting planes. The second method uses an additional inclined double-crystal monochromator cut such that the aberration introduced by the first pair of crystals is nearly completely compensated by the second pair of crystals. This method is independent of wavelength. Both methods are illustrated by ray tracing. PMID- 16714830 TI - Photoemission studies of adsorbates on metal surfaces. AB - A brief review is presented of the general field of synchrotron radiation photoemission of adsorbates on well characterized metal surfaces, contrasting some of the earliest work of the 1970s with more recent experiments, and highlighting some of the likely future developments that may derive from the use of third-generation synchrotron radiation facilities. Four general types of study are identified: shallow core-level photoemission either for the fingerprinting of the chemical state, or for quantitative structural studies via photoelectron diffraction, and valence-level photoemission either to study the two-dimensional band structure of adsorbate-induced surface-localized states, or to investigate the orientation of molecular adsorbates through the use of symmetry selection rules. PMID- 16714832 TI - Tests of an asymmetric monochromator to provide increased flux on a synchrotron radiation beamline. AB - Asymmetric channel-cut monochromators have been tested at the SRS. Results from both focused and unfocused beamlines have shown a threefold improvement in flux when compared with the flux obtained from a symmetric cut Si(111) monochromator. Some problems with using such monochromators and possible modifications are described. PMID- 16714833 TI - Focusing of X-rays by Total External Reflection from a Paraboloidally Tapered Glass Capillary. AB - The first observation of a true geometrical focus of X-rays well beyond the exit of a paraboloidally tapered glass monocapillary is reported. An intensity gain of 250 +/- 20 into a 6 x 9 mum pinhole for 8 keV X-rays and transmission efficiencies of more than 90% below 20 keV were observed. PMID- 16714834 TI - Synchrotron radiation laue diffraction for the time-resolved study of a transformation in crystals of p(4)n(4)cl(8). AB - Experiments are described to show some of the potential of the synchrotron radiation Laue method for the study of structural change within single crystals. In the metastable tetragonal crystals of P(4)N(4)Cl(8) the eight-membered P(4)N(4) ring is in a boat conformation, with symmetry {?bar 4}. On heating to ca 340 K the crystals transform, slowly to a second tetragonal form in which the ring conformation is a chair, its symmetry {?bar 1}. Both structures are known [Hazekamp, Migchelsen & Vos (1962). Acta Cryst. 15, 539-543; Wagner & Vos (1968). Acta Cryst. B24, 707-713]. In the transformation the molecular packing, unit-cell dimensions and crystal quality remain almost unchanged. To study this transformation, series of Laue diffraction patterns were recorded at 2-3 min intervals over a period of 30-40 min, while the temperature was raised to 373 K. For two series, reflection intensities were measured and they allowed determination and refinement of the fraction of boat and chair molecules present in a mixed boat/chair model of the structure. No significant change in the crystal occurs below ca 340 K; at or above 340 K, 40-50% of the molecules are converted from boat to chair conformations within 5 min, but the remainder of the conversion is much slower, even when the temperature is raised towards 370 K. PMID- 16714835 TI - Microfocus X-ray Diffraction of Spherulites of Poly-3-hydroxybutyrate. AB - The microfocus X-ray beamline at the European Synchrotron Radiation Facility has been used to investigate the variation in molecular orientation and crystallinity in spherulites of the organic polymer poly-3-hydroxybutyrate (PHB). This is the first report of the correlation of optical and X-ray measurements on spherulitic polymer films where X-ray diffraction patterns have been recorded and displayed continuously in real time while the specimen was tracked in steps of 10 mum across an incident X-ray beam with a diameter as small as 10 mum. PMID- 16714836 TI - Photoemission Study of CO Adsorption on Gd. AB - Angle-resolved UV photoemission spectroscopy has been used to investigate the interaction of CO with Gd films, grown on W(110). The results suggest the CO absorbs dissociatively, initially forming Gd(2)O(3), with subsequent catalytic oxidation of CO to form carbonate. PMID- 16714837 TI - Sample housing for high-energy Compton measurements. AB - In order to obtain reliable Compton spectra using high-energy photons, the scattered beam must be free of parasitic photons, because of the weakness of the double-differential inelastic cross section. The purpose of the device presented in this paper is to remove photons scattered by air, the sample holder or any other stopper. It has been tested on ESRF beamline 2, which is devoted to materials science. PMID- 16714838 TI - Prices of IUCr journals. PMID- 16714839 TI - A case of hypothyroid Graves' disease following external radiation therapy to the cervical region. AB - A case of hypothyroid Graves' disease occurred following external radiation therapy to the cervical region is described. Severe hypothyroidism developed in a 56-year-old man 6 months after external radiation therapy for submandibular cancer. Serological evaluation of thyroid autoimmunity revealed the presence of antithyroid antibodies and thyrotropin-binding inhibitory immunogloblins (TBII). Diplopia, limitation of downward gaze, and palpebral edema developed 2 years after levothyroxine replacement therapy. Ocular magnetic resonance imaging revealed marked hypertrophy of the bilateral extraocular muscles with signal hyperintensity on T2-weighted images. This infiltrative ophthalmopathy showed marked improvement after additional treatment with high-dose methylprednisolone and orbital radiation, in parallel with a decrease in TBII. These results suggest that radiation-associated thyroidal injury might be associated with the etiology of hypothyroid Graves' disease. PMID- 16714840 TI - Relationship between the serum concentrations of C-reactive protein and parameters of adiposity and insulin resistance in patients with type 2 diabetes mellitus. AB - Serum C-reactive protein (CRP) concentrations have been reported to be associated with body fat, especially visceral fat accumulation, but most studies up to now have been conducted on non-diabetic subjects. In this study, we investigated the association between the serum CRP concentrations and parameters of adiposity and insulin resistance in both Japanese type 2 diabetes patients and non-diabetic subjects. A total of 248 Japanese subjects (140 type 2 diabetes patients and 108 non-diabetic subjects) were enrolled for the study. The degree of insulin resistance was estimated by the homeostasis model assessment (HOMA-R) method. Fat accumulation was evaluated by measuring visceral and subcutaneous fat areas at the level of the umbilicus in abdominal CT scans. To assess hepatic fat content, the ratio of CT attenuation value of the liver to that of the spleen (L/S ratio) was calculated. Serum CRP was found to be significantly correlated with various indices of adiposity, including L/S ratio, visceral fat area (VFA), subcutaneous fat area (SFA), and HOMA-R, in both the diabetic patients and the non-diabetic subjects. After adjustment for five variables (age, gender, serum CRP, HbA1c, and smoking), serum CRP was still significantly correlated with L/S ratio, VFA, SFA, and HOMA-R in the diabetic patients. We also found that changes in serum CRP concentrations were correlated with changes in the VFA and SFA at 1 year after the baseline in 24 diabetic patients. We conclude that serum CRP may be closely related to the degree of liver steatosis and visceral fat accumulation in Japanese type 2 diabetes mellitus patients. PMID- 16714841 TI - Severe hypothyroidism induced by thyroid metastasis of colon adenocarcinoma: a case report and review of the literature. AB - An 85-year-old man who had undergone a right hemicolectomy for colon cancer presented with severe hypothyroidism and hoarseness 21 months after the operation. The serum thyrotropin (TSH) was markedly elevated to 118.14 microIU/mL and serum free thyroxine (fT4) level was markedly suppressed to 0.34 ng/dL. Symptoms of hoarseness and neck swelling were already evident 4 months prior at which time tests for normal thyroid function were performed. The patient was referred due to aggravated pain on his diffusely enlarged hard goiter. An enlarged thyroid with some calcification was noticed in the neck ultrasonography with multiple cervical lymphadenopathies. Core biopsy of the thyroid gland showed invasion of poorly differentiated adenocarcinoma cells. Immunohistochemical studies showed positive staining only for carcinoembryonic antigen (CEA). There were multiple lung parenchymal nodules and adrenal masses at the time of evaluation. The patient was started on palliative chemotherapy with thyroid hormone replacement and gradually became euthyroid. From these findings and the clinical observations, thyroid metastasis with hypothyroidism developing acutely from metastatic colon adenocarcinoma was diagnosed. PMID- 16714842 TI - Neonatal exposure to diethylstilbestrol alters the expression of DNA methyltransferases and methylation of genomic DNA in the epididymis of mice. AB - Fetal and neonatal exposure to diethylstilbestrol (DES) is known to cause many abnormalities, such as cancer, in the male and female reproductive tracts later in life, and epigenetic mechanisms, such as DNA methylation, may be involved in these processes. In the present study, newborn C57BL/6 male mice were exposed to 3 mug of DES from postnatal days 1 to 5. Subsequently, the expression levels of the DNA methyltransferases Dnmt1, Dnmt3a and Dnmt3b and the transcription factors Sp1 and Sp3, which have been reported to regulate the expression of Dnmts, were examined at days 5, 14 and 30. Furthermore, restriction landmark genomic scanning (RLGS), which can analyze genome-wide DNA methylation, was performed to clarify whether or not aberrant DNA methylation was present in the epididymis of the DES treated mice at day 30. Increased expression of Dnmt3b was observed at days 5 and 14, followed by increased expression of Dnmt1 and Dnmt3a at day 30, as evaluated by real-time RT-PCR. The expression of Sp1 was also increased at day 30. The RLGS analysis revealed that 7 loci of the genomic DNA were demethylated and 1 locus was methylated in the epididymis of the DES-treated mice. Four of these loci specifically demethylated in DES-treated mice were cloned, and all were found to be located within CpG islands near genes. In conclusion, our results indicated the possibility that DES-induced abnormalities of reproductive organs are associated with altered expression levels of DNA methyltransferases and DNA methylation. PMID- 16714843 TI - Phenotypic and molecular characteristics of Escherichia coli isolated from aquatic environment of Bangladesh. AB - Pathogenic Escherichia coli remains important etiological agent of infantile diarrhea in Bangladesh. Previous studies have focused mostly on clinical strains, but very little is known about their presence in aquatic environments. The present study was designed to characterize potentially pathogenic E. coli isolated between November 2001 and December 2003 from aquatic environments of 13 districts of Bangladesh. Serotyping of 96 randomly selected strains revealed O161 to be the predominant serotype (19%), followed by O55 and O44 (12% each), and 11% untypable. Serotype-based pathotyping of the E. coli strains revealed 47%, 30%, and 6% to belong to EPEC, ETEC, and EHEC pathotypes, respectively. The majority of the 160 strains tested were resistant to commonly used antimicrobial agents. Plasmid pro-filing showed a total of 17 different bands ranging from 1.3 to 40 kb. However, 35% of the strains did not contain any detectable plasmid, implying no correlation between plasmid and drug resistance. Although virulence gene profiling revealed 97 (61%) of the strains to harbor the gene encoding heat stable enterotoxin (ST), 2 for the gene encoding Shiga toxin (Stx), and none for the gene for heat-labile enterotoxin (LT), serotype-based pathotyping of E. coli was not fully supported by this gene profiling. A dendrogram derived from the PFGE patterns of 22 strains of three predominant serogroups indicated two major clusters, one containing mainly serogroup O55 and the other O8. Three strains of identical PFGE profiles belonging to serogroup O55 were isolated from three distinct areas, which may be of epidemiological significance. Finally, it may be concluded that serotype-based pathotyping may be useful for E. coli strains of clinical origin; however, it is not precise enough for reliably identifying environmental strains as diarrheagenic. PMID- 16714844 TI - A legionellosis case due to contaminated spa water and confirmed by genomic identification in Taiwan. AB - Tracing the source of a legionellosis (LG) case revealed that the Legionella pneumophila (LP) strain isolated from patient's sputum shared the same serogroup (SG) and PFGE-type with 4 LP strains obtained from a spa center. With a high LP contamination rate (81.2%, 13/16) in all of its 16 basins, this spa center was also found to have a multi-genotypic distribution among its 13 LP isolates, which can be categorized into 5 PFGE-types. Despite such a serious contamination in the spa center, which usually had ca. 100 visitors per day, this male patient, bearing LG-risk factors of long-term heavy smoking and alcoholism, was the only case identifiable after an active investigation. To explore the possible reason for this sporadic infection, all 5 PFGE-types of LP isolated were assayed for their presence of two important virulent genes (lvh and rtx A) and were identified as either less-virulent (lvh (+) , rtx A(+)) or non-virulent (lvh (-), rtx A (-)) types. The strong virulent type (lvh (+), rtx A (+)) usually seen in clinical strains elsewhere was not found here. Moreover, the LG-causative type in this infection was the only one to be classified as the less-virulent type, with the presence of lvh gene indicating its relatively more virulent potential than other 4 PFGE-types. Accordingly, mutual interaction between LP's virulent potential and patient's health-status was suggested to be the force directing the opportunistic infection of this sporadic case. This is the first spa-associated infection caused by SG 2 of LP. PMID- 16714845 TI - Rapid detection and quantification of Japanese encephalitis virus by real-time reverse transcription loop-mediated isothermal amplification. AB - We established a rapid, quantitative real-time reverse transcription loop mediated isothermal amplification (RT-LAMP) assay targeting the envelope gene of Japanese encephalitis virus. The RT-LAMP enabled us to detect the target product within 1 hr by only reacting reverse transcriptase and Bst DNA polymerase in a single tube at an isothermal temperature. The detection sensitivity of the RT LAMP for Japanese encephalitis virus was 1 PFU, similar to that of conventional reverse transcription-polymerase chain reaction (RT-PCR). Flaviviruses of the Japanese encephalitis virus group, such as Dengue virus and West Nile virus, could not be detected. This confirmed the specificity of the RT-LAMP assay for Japanese encephalitis virus. A standard curve was constructed by plotting viral titer against the time for virus detection by the RT-LAMP, validating the quantitative accuracy of the assay. In addition, the amount of virus estimated by RT-LAMP was strongly correlated (r = 0.902) with that determined by plaque assay, a conventional method for virus quantification. These results indicate that the RT-LAMP assay established in this study is specific for Japanese encephalitis virus, and allows more rapid detection and quantification of the virus. PMID- 16714846 TI - Vaccination with live Escherichia coli expressing Brucella abortus Cu/Zn superoxide-dismutase: II. Induction of specific CD8+ cytotoxic lymphocytes and sensitized CD4+ IFN-gamma-producing cell. AB - Previously we reported that immunization with Escherichia coli DH5alpha expressing Brucella abortus Cu/Zn superoxide dismutase [E. coli (pBSSOD)] induces a protective immune response in BALB/c mice. Here we studied the type of immune defense that the recombinant E. coli induces in mice using as our experimental model Brucella superoxide dismutase Cu/Zn presented by J744.A1 to sensitized lymphocytes as the target of specific lysis or as cytokine inductors. The results indicate that E. coli carrying the Cu/Zn gene was able to induce specific cytotoxic T cells, mainly from CD8(+) subpopulation and IFN-gamma-producing cells belonging in their vast majority to the CD4(+) subpopulation. PMID- 16714847 TI - Quorum sensing system lactones do not increase invasiveness of a MexAB-OprM efflux mutant but do play a partial role in Pseudomonas aeruginosa invasion. AB - We studied the quorum sensing (QS) system and the related homoserine lactones (HSLs) observing Pseudomonas aeruginosa invasion using the epithelial cell monolayer penetration assay model. Compared to the PAO1 wild-type, the QS mutants, DeltalasI and DeltarhlI, were compromised in their capacity to invade. The decreased invasiveness of DeltarhlI was restored by adding 100 microM exogenous C(4)-HSL. However, the decreased invasiveness of an efflux mutant, DeltamexAB-oprM, was not restored in the presence of exogenous HSLs. The QS system partially plays a role in P. aeruginosa invasion; however, C(4)-HSL and 3 O-C(12)-HSL are not the essential determinants for invasiveness for P. aeruginosa. PMID- 16714848 TI - First detection of Rickettsia in soft-bodied ticks associated with seabirds, Japan. AB - Rickettsia was first detected in seabird soft-bodied ticks, Carios capensis and C. sawaii in Japan. According to sequence analysis, Rickettsia in Japan was identical to Rickettsia scc31 in C. capensis in the U.S.A. This suggested that an environmental circulation had consisted among microorganisms, ticks and long distance migratory seabirds around the Pacific Ocean. PMID- 16714849 TI - Presence of LuxS/AI-2 based quorum-sensing system in Vibrio mimicus : luxO controls protease activity. AB - Presence of the quorum-sensing regulation system in Vibrio mimicus was investigated. The culture supernatants of V. mimicus strains were found to possess AI-2 autoinducer like activity, and the strains were found to harbor the genes which are homologous to luxS, luxO, and luxR of V. harveyi. These genes of V. harveyi have been shown to be important components of V. harveyi-like quorum sensing system. The luxO gene homologue known to encode LuxO, the central component of the regulation system, was disrupted, and effects on protease and hemolysin activity were studied. Disruption of luxO gene resulted in the increased protease activity, but the hemolysin activity did not vary considerably. PMID- 16714851 TI - Endoscopic stent insertion for the palliation of malignant gastric outlet obstruction. AB - BACKGROUND/AIMS: Controversy exists as to whether endoscopic or minimally invasive/open bypass is superior in cases of malignant gastric outlet obstruction. METHODS: Retrospective observational study following those patients admitted to the hospital between January 1999 and December 2004 (n = 23) with gastric outlet obstruction. In all suitable patients endoscopic stent insertion was attempted. Symptomatic improvement was monitored as was time until discharge as well as need for readmission. We also recorded morbidity and mortality associated with the procedure as well as final survival in days. RESULTS: Stents were inserted successfully in 21 patients. Two patients went on to have a laparoscopic bypass. Stent placement relieved obstructive symptoms in all patients. A median inpatient stay of 4 days was achieved (range 2-9 days). One patient was admitted with recurrent symptoms 9 months following his initial treatment. Investigation showed the distal end of the stent to be occluded by further tumor. His symptoms were relieved by the insertion of a second stent. In 19 patients death was due to metastatic disease. Operative mortality: cerebrovascular accident day 2 (n = 1), duodenal perforation day 4 (n = 1). CONCLUSION: Patients with malignant gastric outlet obstruction not suitable for resection should be assessed for insertion of an enteral stent. PMID- 16714850 TI - Susceptibility of Mycobacterium tuberculosis strains to gatifloxacin and moxifloxacin by different methods. AB - BACKGROUND: Considering the potentials of gatifloxacin and moxifloxacin in the treatment of tuberculosis, the present study was aimed to define resistance to both these drugs. METHODS: Fifty Mycobacterium tuberculosis isolates, consisting of 30 ofloxacin-susceptible and 20 ofloxacin-resistant strains, were tested for their susceptibility to gatifloxacin and moxifloxacin using different susceptibility testing methods, namely the absolute concentration method on Lowenstein-Jensen medium (LJ), the proportion susceptibility testing method (PST) on LJ and 7H11 agar media, and the BACTEC radiometric method. RESULT: The minimal inhibitory concentration (MIC) of gatifloxacin and moxifloxacin was 1 microg/ml by the absolute concentration method on LJ. In the PST method on LJ and 7H11, using a criterion of > or =1% growth as resistant, there was 100% agreement with the absolute concentration method at a concentration of 0.5 microg/ml for gatifloxacin, and 96% agreement with the BACTEC method at a concentration of 0.25 microg/ml. For moxifloxacin, results by the PST method showed 96% agreement with the absolute concentration method on LJ at a concentration of 1 microg/ml and 92% agreement at a concentration of 0.5 microg/ml for both the absolute concentration method on 7H11 and the BACTEC method. CONCLUSIONS: The MICs of gatifloxacin and moxifloxacin were much lower than the MICs of other quinolones like ofloxacin and ciprofloxacin. Additionally, these two drugs have shown a low mean MIC and low concentration as a definition of resistance, which might help in treating the patients with low levels of quinolone resistance. PMID- 16714852 TI - Use of the low-dose corticotropin stimulation test for the monitoring of children with asthma treated with inhaled corticosteroids. AB - OBJECTIVE: Subnormal hypothalamic-pituitary-adrenal (HPA) function and rare cases of adrenal crisis have been reported in asthmatic children treated with inhaled corticosteroids. We investigated subnormal HPA activity and followed up affected patients until recovery of normal HPA functions. STUDY DESIGN: 100 children with persistent asthma underwent low-dose corticotropin testing, with the administration of 1 microg of 1-24 ACTH intravenously. Treatments were beclomethasone dipropionate as a metered-dose inhaler, n = 14, budesonide as a dry-powder inhaler, n = 16, fluticasone propionate as a metered-dose inhaler n = 31 or a dry-powder inhaler n = 39. The mean commercially labelled dose was 520 +/ 29 microg/day (mean +/- SEM, range: 160-1,000) and the equipotent dose (which compares the efficiency of these drugs for treating asthma and their responsibility for systemic effects) was 890 +/- 55 microg/day (range: 200 2,000). RESULTS: The mean stimulated cortisol level +/- SEM (and range) of the patient was 482 +/- 12 (148-801), and that of 40 age-matched controls was 580 +/- 12.5 (439-726), (SD = 79). The result was subnormal (more than 2 SD below the mean of the controls) in28 of the 100 patients. One-four stepwise decreases of 10 100% in the daily equipotent doses received by the patients with abnormal low dose corticotropin testing results led to normal results in subsequent low-dose corticotropin testing in 27 retested patients. The mean time interval between two tests was 5 months (range: 2-6 months) and the mean period required for normalization of the test was 13 months (range: 2-21). Only one case of asthma exacerbation and no adrenal crisis were observed over these periods. CONCLUSIONS: Decreasing daily equipotent doses led to recovery of normal HPA function without asthma exacerbation. Thus, a revision of the doses of inhaled corticosteroids used in asthmatic children with a progressive decrease to the consensus recommended doses should decrease the systemic effects of inhaled corticosteroids, while minimizing the risk of asthma exacerbation. PMID- 16714853 TI - Truncated recombinant Dobrava hantavirus nucleocapsid proteins induce strong, long-lasting immune responses in mice. AB - We describe the cloning and expression of Dobrava hantavirus (DOBV) nucleocapsid proteins and a truncated form consisting of the first 118 N-terminal amino acids, and the capacity of these E. coli ICONE 200-expressed recombinant proteins (rNp) to induce a protective immune response against DOBV in mice. As an alternative carrier protein, the outer membrane protein A derived from Klebsiella pneumoniae (rP40) has been coupled to different rNp constructs. All recombinant proteins were found to be highly immunogenic after three immunizations of rNp. The immunizations resulted in the induction of a strong Np-specific IgG response with a predominance of IgG1 over IgG2b and IgG2a, suggesting a mixed Th1/Th2 cell involvement. A specific IgG3 response could not be detected. Mice immunized with recombinant DOBV rNp without rP40 showed lower nucleocapsid-specific antibody responses in comparison with the rP40-conjugated constructs, but all mice were found to be protected against DOBV challenge. Our results indicate that the rNp constructs coupled to rP40, represent promising vaccine candidates. PMID- 16714854 TI - Comparison of two commercially available pp65 antigenemia tests and COBAS Amplicor CMV Monitor for early detection and quantification of episodes of human CMV-viremia in transplant recipients. AB - The performance of two commercially available CMV pp65 antigenemia (AG) assays (CINA kit, Argene Biosoft, CK, and CMV Brite kit, IQ Products, CBK), and a quantitative PCR test (COBAS Amplicor CMV Monitor Test, Roche Diagnostics, CACM) was evaluated with 667 blood samples from 215 transplant recipients. The diagnostic value of the three tests was defined by their ability to detect episodes of systemic CMV activity. A test score was defined, assigning 7 virtual units (VU) for any episode detected first, 5 VU for any episode detected second, and three VU for any episode detected last. Within the evaluated collective, the following overall scores could be determined for the three assays: 45 VU for the CK, 39 for CACM and 31 for the CBK assay. We conclude from our results that (1) the CK is superior to the CBK assay in detecting episodes of CMV antigenemia, and (2) quantitative serum CMV PCR with CACM is not generally superior to pp65 antigenemia testing. PMID- 16714855 TI - Sequential accumulation of the basal core promoter and the precore mutations in the progression of hepatitis B virus-related chronic liver disease. AB - OBJECTIVES: Despite the pathogenic role of the basal core promoter (BCP) and the precore mutations in chronic hepatitis B virus (HBV) infection, their role in the progression of liver disease is still controversial. We analyzed whether the accumulation of these mutations might enhance the progression of HBV-related chronic liver disease. METHODS: Forty consecutive patients at each clinical status were analyzed. Clinical statuses were as follows: HBeAg-positive asymptomatic carrier (HBeAg(+) ASC) (defined as HBeAg(+), anti-HBe(-), HBV-DNA(+) by hybridization, normal ALT); inactive HBsAg carrier; chronic hepatitis B; liver cirrhosis. The genotype and the BCP/precore regions were determined by PCR using genotype specific primers and direct sequencing, respectively. RESULTS: All patients except one were infected with genotype C. The A to T mutation at nucleotide 1762 and/or G to A mutation at nucleotide 1764 were found in 30% in HBeAg(+) ASC, 65.7% in inactive HBsAg carrier, 95% in chronic hepatitis B, and 90% in liver cirrhosis (p < 0.001). The prevalence of the G to A mutation at nucleotide 1896 was 5% in HBeAg(+) ASC, 22.5% in inactive HBsAg carrier, 32.5% in chronic hepatitis B, and 50% in liver cirrhosis, respectively (p < 0.001). The T to C/A mutation at nucleotide 1753 in the BCP and G to A mutation at nucleotide 1899 in the precore were more frequent in liver cirrhosis than in the other clinical statuses (p < 0.05). CONCLUSION: Sequential accumulation of mutations in the BCP/precore has an important role in the progression of HBV-related liver disease. PMID- 16714857 TI - Cytomegalovirus gB genotype and clinical features in Chinese infants with congenital infections. AB - OBJECTIVE: To investigate cytomegalovirus (CMV) glycoprotein B (gB) genotypes and clinical features in Chinese infants with congenital infections. METHODS: Urine samples were obtained from 79 infants with human CMV infection confirmed by quantitative fluorescence polymerase chain reaction (PCR). A fragment of the gB gene was amplified by nested PCR. CMV gB genotyping was carried out by restriction fragment length polymorphism, and 24 samples of the amplified DNA fragments were verified by DNA sequencing. RESULTS: The levels of CMV DNA in symptomatic and asymptomatic infants were 2.95 x 10(5) and 4.5 x 10(3) copies/ml, respectively, with a significant difference (p < 0.001). In all these cases, the most prevalent genotype was gB1 (50.63%), followed by gB3 (21.52%), gB2 (17.72%), and coinfection (10.13%); gB4 was not found. Moreover, gB1 was more prevalent in infants with liver damage (22/32) than in other symptomatic infants without liver damage (8/22, p = 0.019) or asymptomatic infants (10/25, p = 0.030). The homology of CMV gB in the 24 strains amplified as compared with the sequences of prototype strains in GenBank ranged from 97.06 to 99.64%. CONCLUSIONS: The restriction fragment length polymorphism analysis of CMV gB genotypes was definite and reliable. The gB1 genotype is the most prevalent in Chinese infants with congenital CMV disease, especially in those with liver damage, followed by genotypes gB3, gB2, and gB4. PMID- 16714856 TI - Prediction of breakthrough hepatitis due to lamivudine-resistant hepatitis B virus by a sensitive semiquantitative assay using peptide nucleic acids. AB - OBJECTIVE: The aim of this study was to predict breakthrough hepatitis and analyze the dynamics of lamivudine-resistant hepatitis B virus in patients treated with lamivudine. METHODS: Fifty-five chronic hepatitis B patients treated with lamivudine were included. The emergence of YMDD motif mutants was detected by peptide nucleic acid (PNA) mediated PCR clamping with a detection limit of 10(1) YMDD mutants. We then performed a semiquantitative PCR assay of subjects in whom YMDD mutants were detected. This assay detects 10(2.7)-10(7.7) copies of mutant virus per 1 ml of serum. RESULTS: YMDD mutants were detected in 28 (51%) of the 55 patients. Eight patients stopped medication before viral breakthrough. YMDD mutants appeared transiently despite the continuance of lamivudine therapy in 12 patients. In all 8 patients with breakthrough hepatitis, the quantities of YMDD mutants ranged from 10(2.7)-10(4.7) copies/ml in the two to three months before clinical breakthrough. In contrast, in 12 patients without viral breakthrough, there were always less than 10(2.7) copies/ml YMDD mutants. CONCLUSIONS: Lamivudine-resistant viruses sometimes disappear even during lamivudine administration. Our sensitive quantitative assay proved useful for early detection of YMDD mutants and a threshold of 10(2.7) copies/ml is suggested for predicting viral breakthrough. PMID- 16714859 TI - Evaluation of mild head injury in a pediatric population. AB - Approximately 5 million children present to emergency departments, seeking care for head injuries, each year, and 80% of these children are classified as cases of mild head injury. Due to the huge number of patients and low frequency of intracranial lesions in this group, obtaining a computed tomography scan for each and every patient is a significant economic problem. This study was conducted to identify the clinical parameters and the radiographic findings that may be associated with intracranial lesions in children with mild head injury. 421 patients, with a Glasgow Coma Scale score of 15 and without any focal neurological deficit, were studied. Intracranial lesion was noted in 37 cases (8.8%). Sensitivity of a plain radiogram was 43.2%, and specificity was 93%. An intracranial pathology was demonstrated in 28.9% of the patients with a linear skull fracture. The only clinical parameters associated with an increase in the frequency of detection of intracranial lesions were posttraumatic seizures and loss of consciousness. Age, sex, headache, vomiting and scalp lacerations were not associated with a higher frequency. Even when patients with a history of loss of consciousness or posttraumatic seizure were subtracted from the study group, intracranial lesions were noted in 4.1% of the cases, and in 1.8% neurosurgical intervention was required. Computed tomography is the gold standard in the evaluation of pediatric patients with mild head trauma, and every child who has experienced a head injury should undergo a cranial computed tomography evaluation, even if he or she appears in perfect health. PMID- 16714860 TI - Do grip and pinch strength predict neurologic complications in children with spina bifida and hydrocephalus? AB - BACKGROUND: Neurological complications occur commonly in children with meningomyelocele and can cause significant morbidity and mortality. An earlier study suggested that acute changes in grip and pinch strength could be used to identify individuals at increased risk for developing a neurological complication. OBJECTIVE: To evaluate the use of grip and pinch measurements to screen for neurological problems in children with spina bifida and hydrocephalus. DESIGN: A prospective evaluation of screening tests. PATIENTS: 92 children, born since 1976, who had meningomyelocele and hydrocephalus and were treated at the University of Rochester Medical Center. METHODS: Grip and pinch strength were measured between July, 1991, and June, 2003. RESULTS: Mean grip and pinch strengths were similar to those found in previous studies of children with meningomyelocele; 58 neurological events occurred in 39 (40%) individuals. These included 31 episodes of ventricular shunt failure and 22 symptomatic tethered cord occurrences. Specificity, sensitivity and likelihood ratios were calculated in multiple ways using different criteria for loss of grip or pinch strength and for interval to neurological event. Sensitivities were low (<0.35) and the highest positive likelihood ratio found, using fall in either lateral pinch with 6 months to neurological event, was 2.3. CONCLUSIONS: Despite previous recommendations, grip and pinch measurements were not helpful when used as a routine screening test for neurological dysfunction for children with meningomyelocele and hydrocephalus. PMID- 16714861 TI - Paradoxical response to chemotherapy in neurotuberculosis. AB - OBJECTIVE: Our objective was to investigate the paradoxical response and the factors responsible for such an uncommon (but known) response to chemotherapy in neurotuberculosis. MATERIAL AND METHODS: Ten children with intracranial tuberculomas/neurotuberculosis were retrospectively studied, who were on regular antituberculous chemotherapy. Any deterioration of the neurological condition, increase in the size of the lesion or appearance of new lesions was studied clinically and radiologically during the follow-up period. RESULTS: An atypical response to chemotherapy was observed from 3 weeks to 1 year of chemotherapy. Four of the 10 children, who were receiving chemotherapy for either post tubercular meningitis (TBM), hydrocephalus (2) or TBM (2), developed multiple granulomas in 3 and hydrocephalus in 1 case. The remaining 6 children who were diagnosed to have granulomas at their presentation deteriorated at the follow-up despite regular chemotherapy. CT scans were repeated in 7 of 10 cases at the time of neurological deterioration, out of which 5 children showed appearance of new granulomas or abscess(es) and 2 showed an increase in the size of their preexisting granulomas. One patient with faintly enhancing granulomas in the posterior fossa responded to an additional use of steroids. The remaining 9 patients required surgical intervention on account of their neurological deterioration. CONCLUSIONS: (1) Paradoxical responses to intracranial tuberculoma/neurotuberculosis can occur at any time even up to 1 year during chemotherapy despite a regular standard antitubercular treatment. (2) New granuloma(s) or abscess(es) may appear in children receiving chemotherapy for TBM during the follow-up. (3) Hydrocephalus may also appear despite a regular chemotherapy in treated TBM cases. (4) Immature faintly enhancing tuberculomas have a more likely chance of resolution with antituberculous chemotherapy and glucocorticoids, while a well-formed and probably large-sized (>3 cm) granuloma may have a risk of paradoxical enlargement. PMID- 16714862 TI - Insurance barriers for childhood survivors of pediatric brain tumors: the case for neurocognitive evaluations. AB - OBJECTIVE: The purpose of the present study was to provide empirical evidence of system-based barriers to psychological services for pediatric brain tumor patients when they are medically indicated. METHOD: Insurance claims data covering 263,866 insured lives during the 1996 fiscal year were pooled from a cross-sectional national sample of adults and their families insured by private insurance companies or self-insured firms. Based on inclusion criteria, records for 209 pediatric brain tumor patients aged 18 and under were extracted and analyzed. Claims data including total amount of payments made on behalf of a member, total length of hospital stays, and total number of unique admissions were recorded for all patients, and current procedural terminology (CPT) codes were analyzed to determine frequency of payment for routinely billed psychological procedures. Results were then compared to the frequency of payment for routinely billed psychological procedures for children with other medical conditions. RESULTS: Results indicate that two of the CPT codes commonly associated with neurocognitive evaluations were reimbursed by these third-party payers for pediatric brain tumor patients during the 1996 fiscal year. Additionally, seven of the CPT codes commonly associated with psychological therapy were also reimbursed. CONCLUSIONS: The present findings provide empirical evidence of system-based obstacles (i.e., lack of third-party reimbursement) for medically indicated psychological services in pediatric brain tumor patients. PMID- 16714863 TI - Aquaporin(s) expression in choroid plexus tumours. AB - OBJECTIVE: It was the aim of this study to investigate the pattern of aquaporin 1 (AQP1) expression in normal and neoplastic choroid plexus, with specific reference to the association with communicating hydrocephalus or liquoral cysts. Second, to infer a new view on the cerebrospinal fluid plexus production and on the etiology of the cysts and communicating hydrocephalus occasionally associated with choroid plexus papillomas. MATERIALS AND METHODS: Nineteen paraffin-embedded specimens, 10 of normal choroid plexus and 9 of choroid plexus tumours, were immunostained with a monoclonal antibody raised against the intracellular C terminal AQP1 epitope. Results were analysed in terms of intensity and intracellular distribution of immunostaining and in terms of number of stained cells; they were considered in light of the clinical association with hydrocephalus or liquoral cysts. RESULTS: AQP1 was heavily expressed in the apical side of the choroid epithelium in normal plexus specimens. Choroid plexus papillomas showed a very heterogeneous pattern of AQP1 expression. Immunostaining was absent in the case of choroid plexus carcinoma. Very strong to strong and diffuse AQP1 expression in large to very large papillomas was associated with liquoral cysts or communicating hydrocephalus. CONCLUSIONS: AQP1 expression characterizes normal choroid plexus and plexus papillomas. Intensity and diffusion of AQP1 expression together with the size of the tumour mass are somewhat predictive of communicating hydrocephalus or liquoral cyst, lesions possibly caused by a disturbance of cerebrospinal fluid homeostasis. PMID- 16714864 TI - A case of type III cystic sacrococcygeal teratoma. AB - We report a rare case of congenital type III cystic teratoma that may be falsely diagnosed as an anterior sacral meningocele. A female newborn baby presented with a long, white-colored 4 x 1 cm coccygeal cyst posterior to the anus. Magnetic resonance imaging (MRI) demonstrated that a 10 x 4 cm elliptical intrapelvic and abdominal mass anterior to the sacrum and coccyx extended inferiorly to the retrorectal space. We intraoperatively found that the cyst consisted of a mostly thin, white wall, but also with small posterior narrow nodules. We suggest that all cysts in the sacrococcygeal area should be excluded from teratoma carefully, with MRI. PMID- 16714865 TI - Mycotic aneurysm. an unusual cause of pyrexia of unknown origin in an immunodeficient infant. AB - Intracranial aneurysms in early childhood are rare. Mycotic aneurysms as a cause of pyrexia of unknown origin has never been reported. We present a 9-month-old immunodeficient boy who had a long history of irregular fever from neonatal period and presented to us with subarachnoid hemorrhage. Investigation revealed a basilar top aneurysm. We present the clinicoradiological profile and discuss the management strategies. PMID- 16714866 TI - Successful chemotherapy for congenital malignant gliomas: a report of two cases. AB - We describe the cases of 2 patients with a congenital malignant glioma that responded to chemotherapy. In the first case, a 2-month-old boy had a conjugate deviation to the right side and nystagmus. A T(1)-weighted gadolinium-enhanced magnetic resonance image showed a large tumor in his right frontal lobe. The tumor was partially resected, and the histological diagnosis was malignant ganglioglioma. The child then underwent 6 cycles of chemotherapy (mainly carboplatin and etoposide), and the residual tumor shrank. The tumor was then partially resected during a second operation, after which the patient underwent 5 cycles of chemotherapy (a combination of carboplatin, etoposide, vincristine, ifosfamide, cisplatin and cyclophosphamide). The tumor has not recurred in more than 8.5 years. In the second case, a 2-month-old boy had bulging of the anterior fontanel. The T(1)-weighted gadolinium-enhanced magnetic resonance image showed a large suprasellar tumor. The tumor was partially resected, and the histological diagnosis was anaplastic astrocytoma. The patient underwent 8 cycles of chemotherapy (MCNU, carboplatin and etoposide) and the tumor has not recurred in more than 7.5 years. Our experience indicates that, if surgical removal and chemotherapy are done aggressively for malignant gliomas in neonates and infants, long-term survival is possible. PMID- 16714867 TI - Third ventricular ependymal cyst presenting with acute hydrocephalus. AB - Ependymal cysts are generally located in the cerebral parenchyma but rarely found in the third ventricle. A 4-year-old boy presented with headache, vomiting, and upward gaze palsy. His consciousness gradually deteriorated in the course of 6 h. A magnetic resonance imaging study disclosed dilation of the lateral ventricle and a cystic mass in the third ventricle. We performed an endoscopic resection of the cyst wall. The cyst originated on the lateral wall of the third ventricle and obstructed the aqueduct. Histological examination confirmed a diagnosis of ependymal cyst. The patient recovered quickly and his headache and nausea disappeared. Third ventricular ependymal cysts are a rare cause of acute hydrocephalus but an important differential diagnosis. Their neuroendoscopic resection can resolve disturbances in cerebrospinal fluid circulation, is useful for cyst wall removal, and appears to be superior to shunt placement. PMID- 16714868 TI - Hypothalamic glioma presenting with seizures. a case report and review of the literature. AB - A rare case of hypothalamic glioma in a 7-year-old boy presenting with seizures and diabetes insipidus is reported. Near total decompression of the hypothalamic glioma was done successfully using subfrontal approach. The patient had a stormy postoperative course due to status epilepticus but went on to make a complete recovery. Postoperative radiotherapy/chemotherapy was not given in view of tumor histology (low grade glioma), patient's age and tumor location. Patient remains symptom and seizure free on antiepileptics at 3-year follow-up. The case is presented in the light of its rarity and the literature is reviewed. PMID- 16714869 TI - Giant hypothalamic hamartoma operated through subfrontal approach with orbitary rim osteotomy. AB - INTRODUCTION: Hypothalamic hamartomas are associated with precocious puberty, gelastic seizures and severe refractory epilepsy. Treatment options include surgical resection, radiofrequency and radiosurgery. CASE REPORT: A 7-month-old girl presented with gelastic seizures and developmental delay related to a giant hypothalamic hamartoma. The patient was operated through a subfrontal approach. Intraoperatively the lesion appeared intimately adherent to the right internal carotid artery. Seizure control was improved after tumoral decompression. CONCLUSIONS: Treatment of giant hypothalamic hamartomas should always include surgical resection, given the mass effect over surrounding vital structures. Subfrontal approach with orbitary rim osteotomy provides a wide exposure with minimal frontal lobe retraction. Close adherence of hypothalamic hamartoma to vascular structures may be present, requiring careful surgical manipulation. PMID- 16714870 TI - Atypical teratoid-rhabdoid tumor spreading along the trigeminal nerve. AB - We here describe the case of a boy with an atypical teratoid-rhabdoid tumor (ATRT) of the 4th ventricle at 1 year of age and a local tumor recurrence at 19 months of age. Due to brainstem infiltration, only incomplete tumor resection was possible each time. High-dose chemotherapy, stem cell transplantation and irradiation resulted in complete tumor remission on a control MRI. At 8 years of age, another tumor appeared extending from the cerebellopontine angle along the right trigeminal nerve through Meckel's cave into the cavernous sinus. The trigeminal tumor was not in continuity with the primary ATRT but was located within the field of prior irradiation, neuroradiologically mimicking a schwannoma or a meningioma. The origin of the trigeminal tumor as a late metastasis of the former ATRT or as a less likely irradiation-induced secondary ATRT and the operative approach are discussed. PMID- 16714871 TI - A case of rudimentary lateral parietal cephalocele: extracranial meningothelial and glial tissue without intracranial communication or bony defect. AB - The authors report a case of a 7-month-old infant with a right lateral parietal scalp lesion intermittently leaking fluid similar to cerebrospinal fluid that histologically demonstrated meningothelial and glial cells. At surgical removal, however, no fibrous stalk or bony defect could be identified connecting the lesion with the intracranial compartment. While the embryologic mechanism of this lesion is unclear, the prognosis for normal neurological development appears excellent. PMID- 16714872 TI - Transverse sinus thrombosis presenting as pseudotumor cerebri. PMID- 16714873 TI - Surgical treatment of subacute epidural hematoma caused by a vacuum extraction with skull fracture and cephalohematoma in a neonate. PMID- 16714874 TI - [Chronic hemorrhagic radiation proctitis treated by sucralfate enema]. PMID- 16714875 TI - [Antibiotic resistance of Helicobacter pylori isolated from Korean patients]. AB - The distribution of minimal inhibitory concentrations (MIC) for amoxicillin, clarithromycin, metronidazole, tetracycline, azithromycin, and fluoroquinolone (ciprofloxacin, levofloxacin, and moxifloxacin) have shifted to higher concentrations from 1987 to 2003 in Helicobacter pylori (H. pylori) strains isolated from Korean patients. MIC values of secondary isolates were higher than those of primary isolates. Of treatment-failure patients, 16.4% showed mixed infections with both antibiotic-susceptible and -resistant H. pylori strains. A total of 89.6% of patients with treatment failure and 52.3% of patients without antibiotic treatment had H. pylori strains resistant to two or more antimicrobial agents (multi-drug resistance, MDR). The most common antibiotics showing MDR were clarithromycin, metronidazole, and azithromycin. The resistance rates to both amoxicillin and clarithromycin were 34.3% in secondary isolates and 6.2% in primary isolates. The resistance rates to both clarithromycin and metronidazole were 73.1% in secondary isolates and 7.7% in primary isolates. In addition, there was a significant difference in antibiotic resistance between two institutions located at Seoul and Gyeonggi provinces. To provide adequate informations about susceptible antibiotics to clinicians, continuous surveillance of antibiotic susceptibilities is needed in Korea. PMID- 16714876 TI - [Cyclo-oxygenase-2 and p53 immunoreactivity in superficial early colorectal carcinoma]. AB - BACKGROUND/AIMS: De novo colorectal carcinoma shows more aggressive behavior including submucosal invasiveness. Both p53 and cyclo-oxygenase-2 (COX-2) have been shown to be involved in colon carcinogenesis, progression from adenoma to carcinoma, and submucosal invasion by tumor. We performed this study to evaluate the expression of p53 and COX-2 protein in de novo carcinoma, compared with ex adenoma carcinoma. METHODS: Twenty three flat adenomas, 19 ex-adenoma carcinomas, 6 de novo carcinomas were included in this study. The expression of p53, COX-2 and Ki-67 were examined immunohistochemically. RESULTS: Both ex- adenoma carcinomas and de novo carcinomas showed similar size and shape. Positive staining for p53 was detected in 3 of 23 (13%) flat adenomas, in 11 of 19 (57.8%) ex-adenoma carcinomas (p < 0.05), and in 1 of 6 (16.6%) de novo carcinomas. Increased numbers of COX-2 positive tumor cells were observed in 1 of 23 (4.3%) flat adenomas, in 2 of 19 (10.5%) ex-adenoma carcinomas, and in 3 of 6 (50%) de novo carcinomas. COX-2 positive expression showed increased tendency in de novo carcinoma (p = 0.073). There was no correlation between COX-2, p53, and Ki-67 expression. CONCLUSION: De novo carcinoma shows increased tendency of COX-2 expression, but decreased p53 expression when compared to ex-adenoma carcinoma. These immunohistochemical findings are in accordance with the fact that de novo carcinoma has no preceding adenoma, with more frequent submucosal invasion despite the small lesion size. PMID- 16714877 TI - [Usefulness of NCI-N87 cell lines in Helicobacter pylori infected gastric mucosa model]. AB - BACKGROUND/AIMS: The unavailability of human gastric cell lines representative of the normal gastric epithelial function such as polarized monolayer restricts the application of cell culture system in approaching the field of Helicobacter pylori (H. pylori) infected gastric mucosa models. The present investigation aimed at assessing the usefulness of NCI-N87 cell line as an adequate cellular model to study the pathophysiology of human H. pylori infection. METHODS: For the identification of epithelial phenotypes at low magnification, cells were observed on a phase-contrast microscope and confocal microscope. Transepithelial resistance (TER) was measured on NCI-N87 cells seeded on Transwell to identify monolayer polarity two or three times a week after confluency. The IL-8 level was determined by ELISA at 24 hours after the administration of HP60190 and IL-1 alpha on NCI-N87 cells. IL-8 level was compared in both upper and lower well with the control. RESULTS: A monolayer phenotype was observed in NCI-N87 cell lines by using confocal microscope. TER was measured as 400-500 (Omega x cm2) at two or three weeks after cell culture. In NCI-N87 cell lines, IL-8 level was significantly increased after 24 hour compared to control, and was prominent in the lower well. CONCLUSIONS: These results suggest that NCI-N87 cell line may be useful in H. pylori infected gastric mucosa model. PMID- 16714878 TI - [Clinical characteristics of gastroesophageal reflux diseases and association with Helicobacter pylori infection]. AB - BACKGROUND/AIMS: The prevalence of gastroesophageal reflux disease (GERD) in Korea was believed to be low until now. Korea is now believed to be on the evolving stage of GERD in its' prevalence. The aims of this study were to evaluate the epidemiologic and clinicopathologic characteristics among the subgroups of GERD i.e. non-erosive GERD (NERD), erosive GERD (ERD) and Barrett's esophagus (BE), and the role of Helicobacter pylori (H. pylori) infection in the pathogenesis of GERD. METHODS: A total of 253 patients with typical symptoms of GERD who underwent EGD were enrolled from October 2002 to January 2004. Patients were grouped as NERD, ERD or BE based on the symptoms and endoscopic findings. BE was histologically confirmed if necessary. Various clinical parameters including the status of H. pylori infection were analyzed. RESULTS: Among 253 patients, 106 patients were classified as NERD while 116 and 31 patients were classified as ERD and BE group respectively. BE and ERD group showed no gender predilection while NERD showed female preponderance (2.31:1, p < 0.05). NERD group were younger (49.57 y.o.) than BE (57.87 y.o.) and ERD (52.30 y.o.) group. About three quarters of the patients of erosive esophagitis were LA-A (74.2%) grade. This suggests the mild nature of erosive esophagitis in Korea. ERD showed significantly higher BMI (kg/m2) compared to NERD (p < 0.05). Hiatal hernia was frequently associated with BE and ERD (p < 0.05), but less frequently in NERD. Overall H. pylori positivity among GERD was significantly lower than the age and gender matched control group (p < 0.05). CONCLUSIONS: Subgroups of GERD in Korea showed different epidemiologic and clinical characteristics. Lower rate of H. pylori infection among GERD group may reflect the protective role of H. pylori infection regarding GERD prevalence in Korea. PMID- 16714879 TI - [Therapeutic efficacy of balloon-occluded retrograde transvenous obliteration in patients with gastric variceal bleeding]. AB - BACKGROUND/AIMS: Although balloon-occluded retrograde transvenous obliteration (BRTO) has been used as a new procedure for gastric variceal bleeding due to its feasibility and minimal invasiveness, reports regarding the results of BRTO are not well presented in Korea. Therefore, we analyzed the results of our experience in recent 39 months. METHODS: Twenty eight patients who received BRTO for primary hemostasis or secondary prevention of gastric variceal bleeding from December 2001 to March 2005 were analyzed retrospectively. RESULTS: Twenty three men and five women were involved, and the mean age was 53.7+/-9.6 years. Technical and clinical success rates were 89.3% and 85.7%, respectively. Follow-up duration was 17.5+/-12.5 months in 23 patients. Gastric varices disappeared in 78.3% and decreased in 21.7%. Relapses occurred in 4.3% of the patients. Preexisting hepatic encephalopathy improved in all 11 patients. Aggravation of ascites, esophageal varices, portal hypertensive gastropathy were observed in 45.8%, 30.4%, 56.5%, respectively. Increased Child-Pugh score (p < 0.001) and decreased albumin concentration (p = 0.002) were observed 3 days after BRTO, but resolved 7 days later. Increased albumin concentration and decreased Child-Pugh score maintained thereafter. Rebleeding occurred in 3 patients which were caused by esophageal varices. Two-year survival rate was 54.6%. Presence of hepatocellular carcinoma (HCC) (p = 0.001) and Child-Pugh grade (p = 0.033) affected the survival, but HCC was the only independent risk factor (p = 0.010, OR = 15.837) in multivariate analysis. CONCLUSIONS: BRTO is an effective therapeutic procedure for primary hemostasis, secondary prevention, and for improving survival in gastric variceal bleeding patients. PMID- 16714880 TI - [Thirteen cases of intrahepatic biliary cystadenoma and cystadenocarcinoma: a single center experience]. AB - BACKGROUND/AIMS: Biliary cystadenoma (BCA) and biliary cystadenocarcinoma (BCACa) are rare cystic neoplasms that usually arise from the liver. We reviewed the clinicopathologic and radiologic findings of 13 cases of intrahepatic biliary cystic neoplasms. METHODS: Seven patients with BCA and 6 patients with BCACa which were pathologically proven within past 10 years were included in this retrospective study. RESULTS: BCA (7 of 7) was more common in female compared to BCACa (4 of 6). Mean age at diagnosis was 53.4 years (BCA) and 58.5 years (BCACa). Abdominal pain (54%) was the most common presenting symptom. Eleven patients (61.5%) exhibited normal liver function profiles and 5 patients (38%) showed elevated levels of serum CA19-9 levels (mean 894.2 U/mL, range: 78.7 2,080). Mean size of tumor was 11.7 cm (range: 5-15). Most frequent radiologic finding was a single cystic mass with septation. BCACa tended to have intracystic solid portion. The cut surface revealed a unilocular or multilocular cystic mass with mucinous contents. Complete surgical excision was done in 12 patients. After the complete resection, recurrence was observed in 1 case of BCACa. CONCLUSIONS: The possibility of biliary cystic neoplasm should be suspected when an intrahepatic cystic lesion with multiseptation or solid portion is noted on imaging study. In addition, complete excision for definite diagnosis and treatment need to be performed. PMID- 16714881 TI - [A case of duodenal diverticulum accompanied with choledochoduodenal and pancreaticoduodenal fistulas]. AB - Choledochoduodenal fistula (CDF) occurring simultaneously with pancreaticoduodenal fistula is extremely rare. CDF has known to be a chronic sequela of cholelithiasis, but it is unknown whether pancreaticoduodenal fistula results from chronic cholelithiasis as well. We report a case of cholelithiasis accompanied with choledochoduodenal and pancreaticoduodenal fistula opening into small suprapapillary diverticulum in a 80-year-old woman. PMID- 16714882 TI - [A case of hepatoid carcinoma of the pancreas]. AB - Hepatoid carcinoma is a primary extrahepatic neoplasm exhibiting features of hepatocellular carcinoma (HCC) in terms of morphology, immunohistochemistry, and behavior. In many cases, tumor cytoplasm is positive for alpha- fetoprotein (alpha-FP) with elevated serum alpha-FP level. Because not all hepatoid carcinomas are associated with alpha- FP overproduction, diagnosis should be made essentially by histological features of the tumor. We present a case of hepatoid carcinoma of the pancreas in a 21-year-old male patient. Abdominal computed tomography and magnetic resonance imaging revealed an inhomogeneously enhanced pancreatic head mass. Serum alpha-FP level was markedly elevated. He underwent pylorus-preserving Whipple's operation. The tumor showed hepatoid and neuroendocrine components simultaneously. The histopathological diagnosis was hepatoid carcinoma associated with neuroendocrine tumor of the pancreas. Seven months after the surgery, the patient is healthy without evidence of recurrence. To date, only 7 cases of hepatoid carcinoma of the pancreas have been reported in the literature, and this is the first case report in Korea. PMID- 16714883 TI - [A case of stump MALT lymphoma after partial gastrectomy]. AB - Gastrectomy is known to be a risk factor for adenocarcinoma in remnant stomach. It is suggested that reflux of bile juice or duodenal secretion to remnant stomach induces atrophic gastritis, intestinal metaplasia, and gastric adenocarcinoma. Malignant lymphoma in remnant stomach after gastrectomy is very rare. Only about thirty cases are reported in the world, and there is no case report in Korea. Gastric MALT (mucosa-associated lymphoid tissue) lymphoma is associated with Helicobacter pylori infection but the mechanism of lymphoma development in remnant stomach is still unknown. We report a case of low grade gastric MALT lymphoma of gastric stump after 10 years from partial gastrectomy. PMID- 16714884 TI - [A case of secondary amyloidosis presenting as massive gastrointestinal bleeding]. AB - Amyloidosis is a disorder characterized by extracellular deposition of amyloid in various tissues and organs. Gastrointestinal manifestations including gastroparesis, constipation, malabsorption, intestinal pseudo-obstruction, and bleeding are common. GI bleeding is a rare initial symptom which can be fatal in some cases. Absence of systemic symptoms and nonspecific endoscopic findings in amyloidosis may make diagnosis difficult. Therefore, amyloidosis-induced GI bleeding should be considered in patients with an obscure hemorrhage. Recently, we experienced a 65-year-old woman who presented with massive hematochezia as a manifestations of amyloidosis. Colonoscopy and SMA angiography showed massive bleeding in the small and large intestine. Colonoscopic biopsy established amyloidosis. We report this case with a review of the relevant literatures. PMID- 16714885 TI - [Establishment of cell line for in vitro study of Helicobacter pylori infection]. PMID- 16714887 TI - Correlation between changes in regional cerebral blood flow and pain relief in complex regional pain syndrome type 1. AB - OBJECTIVE: Analyzing changes in regional cerebral blood flow (rCBF) with SPECT in complex regional pain syndrome type 1 (CRPS 1), formerly known as reflex sympathetic dystrophy, is an optimal method for evaluating effective pain relief. We attempted to investigate the correlation of changes in rCBF with pain relief during treatments of sympathetic blockade and multimodal epidural pain control. CASE REPORT: We describe a patient with severe CRPS 1 in whom conventional treatment failed to relieve the pain. Combined repeated lumbar sympathetic blocks and long-term epidural morphine, bupivacaine, and ketamine administration provided satisfactory pain relief and functional activity recovery. Six normal control subjects having one Tc-99m HMPAO scan each and the patient with CRPS having 3 Tc-99m HMPAO scans (once before treatment and twice at 4 months and 6 months after treatment, respectively). The patient with CRPS showed lower rCBF than normal controls in the left thalamus and higher rCBF than normal controls in the right parietal lobe and left frontal lobe. After subsequent treatment, the subtraction images showed increased rCBF in the left thalamus and decreased rCBF in the right parietal and left frontal lobes. CONCLUSIONS: Tc-99m HMPAO SPECT showed a relationship of rCBF in the thalamus, parietal lobe, and frontal lobe with pain relief. rCBF alterations may provide an indicator for the quality of pain management for neuropathic pains. Subtraction analysis between pre- and posttreatment, by using statistical parametric mapping (version 2), can be used as an objective indicator for the effectiveness of therapy. PMID- 16714888 TI - Metastatic struma ovarii: the burden of truth. AB - A 58-year-old woman was found to have metastatic thyroid cancer in her liver. This was identified when she was investigated for upper abdominal pain and underwent biopsy of hepatic lesions. She had no palpable thyroid nodule and had a normal ultrasound of the thyroid. Previously, both ovaries had been removed because of tumors. The pathologic findings in one of the ovaries could not be recovered because the procedure was more than 40 years ago, when the patient was a teenager. By a process of elimination, a diagnosis of metastatic struma ovarii was established. Treatment of metastatic thyroid cancer from struma ovarii, including removal of the normal thyroid and administration of I-131, is presented. PMID- 16714889 TI - Appearance of intrathymic parathyroid adenomas on pinhole sestamibi parathyroid imaging. AB - OBJECTIVES: Ectopic inferior parathyroid adenomas (PAs) are frequently located in the anterior mediastinum, quite often in the thymus gland. Correct preoperative localization is particularly helpful for surgical planning. Clinical follow up has shown that most intrathymic adenomas were visualized on pinhole views and located closer to the thyroid than expected. We reviewed the typical appearance of intrathymic PA on pinhole views. METHODS: We retrospectively reviewed sestamibi pinhole parathyroid imaging performed on 163 patients with primary hyperparathyroidism and final diagnoses established by surgery, histology, intraoperative PTH monitoring, and clinical follow up. Studies showing focal increased activity in the lower pole region of the thyroid were selected and divided into 2 groups, group A (foci that are visually not separable from the thyroid) and group B (foci that are completely separated from the thyroid), and correlated with the final diagnoses. RESULTS: Of the 163 patients, 102 had 103 clearly abnormal foci in the lower pole region (bilateral lower pole foci in one study). There were 93 foci in group A and 10 foci in group B. Of the 93 foci in group A, there were 80 normally situated inferior PA, 6 descended superior PA, 3 intrathyroidal PA, one hyperplastic parathyroid gland, one thyroid adenoma, one unidentified, and one intrathymic PA. Of the 10 foci in group B, 3 were eutopic inferior PA and 7 were intrathymic PA. CONCLUSION: Focal increased activity completely separated from the lower pole of thyroid (regardless of the distance of separation) on sestamibi pinhole images indicates a high probability of intrathymic parathyroid adenoma. PMID- 16714890 TI - Obscured bone metastases after administration of hematopoietic factor on FDG-PET. AB - We describe interesting imaging features on a FDG-PET scan in a patient with bony metastases from a rare tumor. The lesions were obscured by intense bone marrow activity after the administration of hematopoietic factor at the time of diagnosis. Correlative MRI, CT scan, and pathology findings have also been illustrated. Follow-up FDG-PET scan demonstrated the hypermetabolic lesions. PMID- 16714891 TI - Ruptured vulnerable coronary plaque and Tc-99m sestamibi-based assessment of infarct size. PMID- 16714892 TI - Epicardial fat: an unrecognized cause of artifact on myocardial perfusion imaging. PMID- 16714893 TI - Gangrene after inadvertent intraarterial injection of amphetamine: appearance on a three-phase Tc-99m MDP bone scan. PMID- 16714894 TI - Improved diagnosis of aortobifemoral vascular graft infection by In-111-labeled leukocyte SPECT. PMID- 16714895 TI - Tc-99m red blood cell uptake in positive somatostatin receptor scintigraphy by two accessory spleens. PMID- 16714896 TI - Esophageal leiomyoma: a rare cause of false-positive FDG scans. PMID- 16714898 TI - Choroidal melanoma with hematogenous spread to the liver: F-18 FDG PET/CT findings. PMID- 16714897 TI - F-18 FDG PET imaging in gastric neuroendocrine carcinoma. PMID- 16714899 TI - 'Rim sign' in liver abscesses. PMID- 16714900 TI - Diaphragmatic paralysis simulating pulmonary infection on gallium-67 scan. PMID- 16714901 TI - CT-MIBI image fusion: transmission and emission tomography for localization of bronchogenic carcinoma and nodal metastasis. PMID- 16714902 TI - Scintigraphic images of breast cancer with Tc-99m cysteine. PMID- 16714903 TI - Single F-18 fluorodeoxyglucose positron emission tomography hypermetabolic focus containing metastatic papillary thyroid cancer within a primary scarring adenocarcinoma lung cancer. PMID- 16714904 TI - Renal transitional cell carcinoma diagnosed by FDG-PET in a uremic kidney. PMID- 16714905 TI - Cyanide-induced parkinsonism and F-18 FDG PET/CT findings. PMID- 16714906 TI - Current readings in nuclear medicine. PMID- 16714908 TI - Providing developmentally competent care for adolescents during labor and birth. PMID- 16714909 TI - Practicing out of the box: rituals and threads of evidence. PMID- 16714910 TI - Best practices in perinatal nursing: professional role development for charge nurses. PMID- 16714911 TI - Free radicals in perinatal and neonatal care, part 2: oxidative stress during the perinatal and neonatal period. PMID- 16714912 TI - Establishment of a Somali doula program at a large metropolitan hospital. AB - To meet the unique cultural needs of Somali childbearing women in the United States, a doula support program was implemented at a major midwestern teaching hospital in the spring of 2002. To date, 123 Somali women who have been served by a Somali doula have given birth to live infants at this hospital. Cesarean birth rates in Somali women who had a doula present at birth were lower than the cesarean birth rate for Somali women not attended by a doula. Surveys with the nursing staff indicate that the nurses who have worked with a doula 3 or more times felt more confident caring for Somali women than nurses who have not worked with a doula at least 3 times. Patient feedback indicates positive attitudes about the doula service. Further research needs and suggestions for establishing such a program in other institutions are included. PMID- 16714913 TI - Does relaxation education in anxious primigravid Iranian women influence adverse pregnancy outcomes?: a randomized controlled trial. AB - Maternal anxiety and stress are found to be predictors of adverse pregnancy outcomes, including low birth weight and prematurity. OBJECTIVE: The aim of the study was to determine whether relaxation education in anxious pregnant Iranian women in their first pregnancy affects selected pregnancy outcomes, including birth weight, preterm birth, and surgical delivery rate. SUBJECTS: A total of 110 obstetrically and medically low-risk primigravid women in Iran with a high anxiety level demonstrated by Spielberger's State-Trait Anxiety Inventory were randomly assigned into experimental and control groups. METHOD: In this randomized controlled trial, the experimental group received routine prenatal care along with 7-week applied relaxation training sessions, while the control group received only routine prenatal care. Anxiety and perceived stress were measured by pre-educational and post-educational intervention. Data related to pregnancy outcomes include birth weight, gestational age at birth, and type of delivery. RESULTS: Significant reductions in low birth weight, cesarean section, and/or instrumental extraction were found in the experimental group compared with the control group. No significant differences were found in the rate of preterm birth. CONCLUSION: The findings suggest beneficial effects of nurse-led relaxation education sessions during the prenatal period. This intervention could serve as a resource for improving pregnancy outcomes in women with high anxiety. PMID- 16714914 TI - The effects of rhinitis, asthma, and acute respiratory distress syndrome as acute or chronic pulmonary conditions during pregnancy. AB - Pulmonary complications from both obstetrical and non-obstetrical causes contribute to a mortality rate as high as 80% in the pregnant population. The effect of numerous mechanical and biochemical physiologic alterations during pregnancy can influence the maternal and fetal outcomes in a woman with a pulmonary complication. Progesterone, the primary hormone of pregnancy, is a respiratory stimulant that enhances carbon dioxide release and alters the maternal pH in favor of releasing oxygen to the fetus. During systemic compromise, which may be experienced as an acute asthmatic attack or respiratory distress syndrome, desaturation and carbon dioxide retention ensue. Under these conditions, the fetus is at risk for perinatal hypoxemia. Although prompt recognition and treatment are important to minimize maternal, fetal, and neonatal morbidity and mortality, evidence-based literature regarding critical care techniques that promote optimal obstetrical outcomes is limited. Therefore, a collaborative approach to the care of these women is warranted. In addition to critical care, emergency medicine, and obstetrical nurses, the medical team may include an obstetrician, a perinatologist, a neonatologist, a pulmonologist, an intensivist, and an immunologist. PMID- 16714917 TI - Brain injury in the infant: the old, the new, and the uncertain. AB - Although neonatal brain injury occurs most frequently after a perinatal hypoxic ischemic insult, recently studies have noted that variable causes such as metabolic and reperfusion events can result in, or aggravate, a brain insult. Current data suggest that about 2 to 5 of 1,000 live births in the United States and more so in developing countries experience a brain injury Approximately 20% to 40% of infants who survive the brain injury develop significant neurological and developmental impairments. The resulting impact on the child, family, and society presents a formidable challenge to health care professionals. Although several important insights have been gained in the last several years about the epidemiology, diagnosis, and mechanism of brain injury, management remains mostly a cocktail of controversial trials. This article provides a comprehensive review of the pathology, clinical manifestations, and timely management of infants with brain injury. PMID- 16714916 TI - Relationship of the first suck burst to feeding outcomes in preterm infants. AB - OBJECTIVE: This study examined the relationship between the number of sucks in the first nutritive suck burst and feeding outcomes in preterm infants. The relationships of morbidity, maturity, and feeding experience to the number of sucks in the first suck burst were also examined. METHODS: A non-experimental study of 95 preterm infants was used. Feeding outcomes included proficiency (percent consumed in first 5 min of feeding), efficiency (volume consumed over total feeding time), consumed (percent consumed over total feeding), and feeding success (proficiency >or=0.3, efficiency >or=1.5 mL/min, and consumed >or=0.8). Data were analyzed using correlation and regression analysis. RESULTS AND CONCLUSIONS: There were statistically significant positive relationships between number of sucks in the first burst and all feeding outcomes-proficiency, efficiency, consumed, and success (r=0.303, 0.365, 0.259, and tau=0.229, P<.01, respectively). The number of sucks in the first burst was also positively correlated to behavior state and feeding experience (tau=0.104 and r=0.220, P<.01, respectively). Feeding experience was the best predictor of feeding outcomes; the number of sucks in the first suck burst also contributed significantly to all feeding outcomes. The findings suggest that as infants gain experience at feeding, the first suck burst could be a useful indicator for how successful a particular feeding might be. PMID- 16714920 TI - Eight-fold path to happiness for the cosmetic surgery patient. PMID- 16714919 TI - Oxygen consumption monitoring by oxygen saturation measurements in mechanically ventilated premature neonates. AB - PURPOSE: To assess oxygen consumption VO2) with arterial and venous oxygen saturation SaO2 and SvO2) from blood measurements for continuous monitoring of mechanically ventilated preterm neonates. METHOD: Twelve preterm neonates with gestational ages ranging from 27 to 34 weeks at birth, who were mechanically ventilated and had umbilical arterial and venous lines in place, were investigated. Analyses were performed with blood samples and continuous monitoring of VO2 from SaO2 and SvO2 measurements. RESULTS: Mean VO2 from blood samples was 3.3 mL/(kg min) (+/-2.49), and that from monitor readings was 8.8 (+/ 4.49). Using curve fit analysis to predict the stability of monitor reading, the period from 8 to 28 hours following blood validation was most stable for SvO2 (cubic curve, R=0.5, P<.001). There is a considerable mix of arterial blood in the venous blood, which increased SvO2 and decreased VO2 in the blood samples during first 8 hours of monitoring following blood draws. A multivariate linear mixed model was established for VO2 measurements including related parameters. Ventilatory weaning was associated with decreased VO2. CONCLUSIONS: These findings warrant caution against interpreting VO2 from blood SvO2 and subsequent monitoring readings during mechanical ventilation for preterm neonates. PMID- 16714921 TI - Desmoplastic inflammatory disorders affecting the orbit. PMID- 16714922 TI - The spectrum of orbital Rosai-Dorfman disease. AB - PURPOSE: To describe the spectrum and treatment of orbital Rosai-Dorfman disease and to review previously documented cases. METHODS: Retrospective, interventional case series of seven patients and literature review. RESULTS: Each patient with Rosai-Dorfman disease had unique disease expression requiring aggressive therapy, such as chemotherapy, radiation, and/or surgical excision. One systemically aggressive case presented with intraocular and choroidal invasion, a previously unreported finding. Response to therapy was variable and unpredictable. CONCLUSIONS: Rosai-Dorfman disease, although historically described as benign and self-limiting, may cause significant morbidity and mortality involving multiple organ systems. Available treatment options may not control the disease. Further research and long-term clinical correlation is necessary. PMID- 16714923 TI - Combined external-endonasal approach for complete excision of the lacrimal drainage apparatus. AB - PURPOSE: The purpose of this study was to describe a new surgical technique for the complete excision of the lacrimal drainage apparatus (LDA) that combines external and endoscopic approaches. METHODS: This study involved a noncomparative, retrospective chart review of the clinical and pathological findings of four patients presenting with LDA papillomas who underwent a combined open and endonasal excision of the lacrimal system. RESULTS: Of the four patients, three were male. The mean age at referral was 41 years, and all cases were unilateral. Histopathology revealed two transitional cell papillomas, one squamous cell papilloma, and one combined transitional/squamous papilloma. Epiphora and an external lesion were the main complaints at presentation. Nasolacrimal duct obstruction was present in all four patients. Papilloma virus infection was suggested in two cases and was confirmed in the only patient who had recurrence. CT identified a solid enhancing mass in two cases. The surgical approach in all cases was performed with the patient under general anesthetic supplemented with infiltration of local anesthesia with vasoconstriction. The lacrimal sac was exposed as per an external dacryocystorhinostomy with biopsy collection from the lacrimal sac lumen to confirm the diagnosis prior LDA excision. The superior aspect of the LDA was isolated by using lacrimal probes in each canaliculus to stabilized parallel incisions and careful dissection toward the common canaliculus until they met the medial aspect of the lacrimal sac. The sac was then separated from the periosteum from the medial orbital wall, using sharp dissection. Finally, an endoscopic dissection of the lower end of the nasolacrimal duct released the most inferior aspect of the LDA, allowing the surgeon to pull and excise the complete system from the external wound. CONCLUSIONS: Extensive LDA papillomas required complete excision of the drainage system to prevent recurrence and/or malignant transformation. The use of a combined approach through an open excision of the superior part of the LDA in conjunction with the direct manipulation of the nasolacrimal duct guided by the nasal endoscope facilitates the complete excision of the system for extensive benign lesions. PMID- 16714924 TI - Myobloc for the treatment of benign essential blepharospasm in patients refractory to botox. AB - PURPOSE: A small percentage of cases with essential blepharospasm or hemifacial spasm will become resistant to botulinum toxin A (Botox). We present our experience treating these patients with botulinum toxin B (Myobloc). METHODS: We reviewed all charts of patients in one physician's practice who received botulinum toxin B after becoming refractory to botulinum toxin A. For each treatment session, patients were evaluated for side effects, relief of spasms, and duration of treatment effect. RESULTS: Data were collected on 16 patients and a total of 93 treatment visits. Average total dosage was 3,633 U per treatment session. Mean duration of beneficial effect was 7.3 weeks and was most commonly rated as fair to excellent. Side effects for botulinum toxin B occurred at a higher rate than is typical for botulinum toxin A. Most common side effects were pain on injection (100%), ptosis (32.3%), facial or mouth droop (22.6%), dry mouth (17.2%), dry eye (9.7%), exposure keratitis (7.5%), and blurred vision (5.4%). The rate of occurrence of side effects appears to be dose-dependent. CONCLUSIONS: Botulinum toxin B is a useful treatment for cases that have become refractory to botulinum toxin A. The duration of beneficial effect is shorter than for type A. The side effect profile is similar to that for toxin type A except for pain on injection and the occurrence of dry mouth. Side effects occurred at a higher frequency than with botulinum toxin A, and the pattern suggests that botulinum toxin B spreads more diffusely. Side effects appear to be dosage-related. PMID- 16714925 TI - Isolated medial orbital wall fractures with medial rectus muscle incarceration. AB - PURPOSE: To retrospectively review and analyze cases of isolated medial orbital wall fractures with medial rectus muscle incarceration presenting to a tertiary ophthalmic plastic surgery practice from 1997 to 2005. METHODS: Retrospective chart review and literature review. RESULTS: Nine cases of isolated medial wall fracture with medial rectus muscle incarceration are presented. The most frequently encountered clinical feature was adduction deficit on the affected side. Extraocular motility improved in all patients who underwent surgery, and mean postoperative enophthalmos was minimal. CONCLUSIONS: Isolated medial orbital wall fractures with medial rectus muscle incarceration are rare. Ocular motility abnormalities were the only indication of underlying fracture in the majority of our cases. Clinicians should be alerted to the anticipated presentation of medial wall fractures with incarceration of the medial rectus muscle, including the possibility of a "white eye" and normal abduction of the traumatized eye. PMID- 16714926 TI - Fibrin sealant for Muller muscle-conjunctiva resection ptosis repair. AB - PURPOSE: To determine the safety and efficacy of fibrin sealant for use in Muller muscle-conjunctiva resection ptosis repair. METHODS: This was a retrospective review of a consecutive case series. All patients underwent Muller muscle conjunctiva resection ptosis repair with fibrin sealant used for wound closure. Surgery was performed in a manner similar to a previously described technique, using fibrin tissue sealant rather that suture for wound closure. Postoperative symmetry was defined as MRD1 of each eyelid within 0.5 mm. RESULTS: Muller muscle conjunctiva resection ptosis repair with fibrin sealant used for wound closure was performed on 53 eyelids of 33 patients. There were 27 female patients and 6 male patients. Twenty patients underwent bilateral ptosis repair and 13 patients underwent unilateral ptosis repair. Average follow-up was 17 weeks (range, 3 to 45 weeks). Mean preoperative MRD1 was 1.22 mm (range, -1.5 to 2.5 mm) in the right upper eyelid and 1.50 mm (range, 0 to 2 mm) in the left upper eyelid. Mean postoperative MRD1 was 3.11 mm (range, 2 to 4.5 mm) in the right upper eyelid and 3.12 mm (range, 1 to 4.5 mm) in the left upper eyelid. Postoperative symmetry was found in 32 of 33 patients (97%). We found no evidence of keratopathy or other complications attributable to the fibrin sealant. CONCLUSIONS: Muller muscle conjunctiva resection ptosis repair with fibrin sealant used for wound closure may allow for predictable results with few complications and appears to be an acceptable alternative to traditional suture techniques. PMID- 16714927 TI - Hyaluronic acid gel (Restylane) filler for facial rhytids: lessons learned from American Society of Ophthalmic Plastic and Reconstructive Surgery member treatment of 286 patients. AB - PURPOSE: To review injection techniques and patient satisfaction with injection of Restylane in various facial areas by American Society of Ophthalmic Plastic and Reconstructive Surgery members. METHODS: Data from 286 patients treated with Restylane in nine American Society of Ophthalmic Plastic and Reconstructive Surgery practices were abstracted to a spreadsheet for analysis. RESULTS: Nine practices performed Restylane injections for 8.8 months on average (range, 2 to 28 months). Average practice volume per patient was 1.2 ml (range, 0.7 to 2.1 ml). Nine of nine practices injected the nasolabial and melolabial folds, 9 of 9 practices injected the lips, and 6 of 9 injected the glabella. Only 2 of 9 practices injected other fillers concurrently. Botox was injected concurrently by 8 of 9 practices. On a scale of 1 to 10, physicians rated average patient discomfort during Restylane injection 4.6 with topical anesthesia and 2.1 with injectable lidocaine, with or without topical anesthesia. The end point for injection was determined by visual cues, volume of injection, extrusion of the product, and palpation. "Problematic" complications, including bruising, swelling, bumpiness, and redness each had an incidence of 5% or less. Patient satisfaction on a scale of 1 to 10 had an average rating of 8.1, compared with that of Botox injection (8.9), upper blepharoplasty (8.9), and collagen injection (6.6). The source of Restylane patients was estimated to be existing Botox patients (45%); existing non-Botox patients (18%); word of mouth (14%); and new patients for other services (13%). CONCLUSIONS: Injection techniques, volume, end points, and anesthesia vary for different facial areas and between practices. Patients experience mild to moderate injection discomfort that is lessened with injectable lidocaine. Self-limited problems occur in about 5% of patients. Physician-determined patient satisfaction is perceived to be higher than that of collagen injection but slightly lower than that of botulinum toxin injection. The major source of Restylane patients was from existing practice patients, especially botulinum toxin patients. PMID- 16714928 TI - Anterior lamella actinic changes as a factor in involutional eyelid malposition. AB - PURPOSE: We conducted a noncomparative, retrospective chart review of 45 patients and 51 eyelids with the diagnosis of involutional entropion or ectropion that underwent full-thickness lower eyelid shortening between June 2001 and February 2004, in whom the severity of actinic damage was analyzed in relation to the eyelid position. Patients with any different surgical approach or other primary causes of abnormal eyelid position, such as paralytic, congenital, or mechanical factors, were excluded. METHODS: After excision, all eyelid specimens were examined by a single anatomic pathologist, who was masked to the type of eyelid malposition. The extent of dermal actinic change was evaluated under light microscopy, according to a previously validated grading system. RESULTS: Fifty one eyelids from 26 male and 19 female patients were analyzed. The mean age at the surgery was 76 +/- 10 years (range, 52 to 92 years), affecting one side in 39 cases and both sides in 6 cases. The most frequent eyelid malposition was ectropion, which affected two thirds of the cases (35 eyelids). Half of the patients presented with mild actinic skin changes; however, the severity of the histologic skin actinic changes was significantly worse in patients with ectropion in comparison to those with entropion (p < 0.0001). CONCLUSIONS: Actinic damage affecting the anterior lamella of the lower eyelid contributes as an additional factor in final eyelid position in patients with involutional eyelid changes. More severe and extensive actinic changes were present in eyelids with ectropion. PMID- 16714929 TI - Histologic comparison of autologous fat processing methods. AB - PURPOSE: To perform a quantitative analysis of adipocyte viability after fat processing during autologous fat transfer, comparing the processing methods of washing, centrifuging, and sedimentation. METHODS: An experimental study was conducted in which 24 fat samples were obtained after processing from 22 patients undergoing autologous fat transfer. Histologic analysis of periodic acid-Schiff stained specimens was then performed. RESULTS: Cell counts per high-powered field of intact adipocytes and nucleated adipocytes and adipocyte cross-sectional area were significantly greater in samples processed by sedimentation, compared with those by centrifuging or washing. CONCLUSIONS: Of the various processing techniques currently used during autologous fat transfer, sedimentation appears to yield a higher proportion of viable adipocytes than does washing or centrifuging. PMID- 16714930 TI - Ocular adnexal pseudo-cyst formation as a characteristic feature of perineural spread in squamous cell carcinoma. AB - PURPOSE: To report the clinical features in a series of patients with perineural spread of squamous cell carcinoma involving periorbital nerves that presented with clinical and/or imaging evidence of pseudo-cystic transformation along the involved nerves. METHODS: A noncomparative, retrospective chart review of the clinical and imaging findings of patients attending a regional orbital surgery department between 1998 and 2005, presenting with a pseudo-cystic orbital mass on clinical examination and/or imaging, which proved to be due to perineural squamous cell carcinoma on histopathology. RESULTS: The study included 8 male patients with a mean age at referral of 66 +/- 11 years. All cases had associated cutaneous squamous cell carcinoma involving the face or scalp, and, in 4 cases, the primary tumor could be identified in the vicinity of the affected orbit. The duration of the symptoms varied from 5 weeks to 9 years (24 +/- 36 months). Altered sensation, including formication and hypo-esthesia in the V1 and V2 trigeminal division, motor nerve palsies, and ptosis in conjunction with a palpable periorbital mass, were the most common presentations. The cystic tumor deposits were assessed histologically with both hematoxylin and eosin and immunoperoxidase stains (S100 protein for neural structures identification and MNF116 as a keratin marker). This demonstrated malignant squamous epithelium both within and around the wall of the tumor deposit and, in continuity, within the nerve running through the lesion. Some nerves showed substantial areas of fibrosis, representing obliteration of the nerve structure caused by involvement by tumor. Treatment modalities included surgical debulking, exenteration, radiotherapy, and combined chemo-radiotherapy. Mean follow-up was 29 +/- 23 months. Eight patients (87.5%) remain alive, and five of them show no evidence of disease. One patient died after progression of the malignancy. CONCLUSIONS: The presence of a cystic lesion in association with sensory or motor deficit in the periorbital region should suggest a diagnosis of perineural spread from a cutaneous squamous cell carcinoma. Because these patients may present to the ophthalmologist first, an awareness of this entity is of critical importance to avoid the delayed diagnosis seen in half of our cases. PMID- 16714931 TI - Orbital complications of hydrogel scleral buckles. AB - PURPOSE: Hydrogel is a biocompatible osmotic expansile material used in contact lenses, scleral buckles, and orbital expanders. Three unique complications from hydrogel scleral buckle use are presented: orbital cellulitis mimicry, fornical shortening with orbital prosthetic intolerance, and orbital pseudotumor. METHODS: A retrospective chart review was conducted on three subjects with unknown hydrogel scleral buckle complications at a tertiary care oculoplastics practice. Demographics, imaging studies, and pathologic specimens were reviewed. RESULTS: All three patients had a history of encircling hydrogel scleral buckle placed for retinal detachment. The mean time from implantation to presentation was 10.7 years (range, 7 to 15 years). One patient had an onset of presumed orbital cellulitis caused by an enlarged buckle and no abscess. The second patient had inability to wear an evisceration prosthesis as the result of a shortened fornix from an enlarged hydrogel scleral buckle implant. The third patient presented with an orbital mass caused by the enlarged implant. In all cases, surgical removal of the expanded buckle relieved the patient's symptoms. CONCLUSIONS: Although the hydrogel scleral band is designed to expand in a controlled fashion, overexpansion can occur. Expansion can cause buckle extrusion, prosthetic intolerance, a pseudo-orbital cellulitis, or orbital pseudotumor and may be due to a chemical change in the hydrogel polymer. Patients with hydrogel scleral buckles should be followed long-term for possible complications. Patients with newer uses of hydrogel, such as orbital expanders, should also be observed for long-term complications. PMID- 16714932 TI - Characterization and localization of the supraorbital and frontal exits of the supraorbital nerve in Chinese: an anatomic study. AB - PURPOSE: To elucidate the characteristics and location of the supraorbital and frontal exits of the supraorbital nerve in Chinese skulls and to compare these findings with other ethnic populations. METHODS: The anatomy of the supraorbital nerve exits was studied in 97 adult Chinese skulls (194 orbits). The characteristics and location of the supraorbital nerve exits were determined. Analyses were performed to compare differences between sexes and sides. Data were then compared with those of the other populations. RESULTS: Supraorbital exits were found in all 194 orbits. Among these, 89 (45.9%) were foramina. Frontal exits were found in 69 of the 194 (35.6%) orbits in which 18 were foramina (9.3%). The location of the supraorbital exits showed a skewed distribution toward the medial aspect of the superior orbital rim. The median distance between the supraorbital exits and the facial midline was 24.56 mm. Eighty percent of the supraorbital exits lay between 20.77 mm and 30.52 mm from the facial midline. The mean frontal exit was 7.02 mm medial to the supraorbital exits. Compared with other populations, a higher proportion of Chinese have foramina above the superior orbital rim as the exits for the supraorbital neurovascular bundle. The location of the supraorbital nerve exit is highly variable in all studied populations. CONCLUSIONS: In a significant proportion of Chinese, the supraorbital nerve exits are located toward the medial aspect of and above the superior orbital rim. Extra care should be taken during dissection around this region to avoid damage to the supraorbital nerve. PMID- 16714934 TI - Periorbital edema as the initial presentation of T-cell prolymphocytic leukemia. AB - A 57-year-old woman presented with a history of progressive bilateral upper and lower eyelid edema. Laboratory tests revealed T-cell prolymphocytic leukemia. Despite systemic treatment, she died 2 weeks after presentation. This life threatening disorder should be added to the differential diagnosis of eyelid edema. PMID- 16714933 TI - Amaurosis after lower eyelid laser blepharoplasty. AB - The transconjunctival CO(2) laser approach to lower eyelid blepharoplasty was used to treat a 66-year-old man presenting with bilateral lower eyelid herniated fat without excess skin. Early postoperative examination revealed left eye blindness. This report presents clinical circumstances of this rare complication and further discusses the most likely causative factors. PMID- 16714935 TI - Multiple eyelid defects in cutaneous Langerhans cell histiocytosis. AB - Langerhans cell histiocytosis is a disorder characterized by the proliferation of histiocytes of the mononuclear phagocyte system. Cutaneous Langerhans cell histiocytosis can develop as a solitary lesion or more frequently as part of a multisystem disease. This report describes a case of cutaneous Langerhans cell histiocytosis affecting the eyelids, resulting in a coloboma-like defect. The clinical features and management are discussed. PMID- 16714936 TI - Discoid lupus erythematosus presenting as unilateral blepharitis. AB - A 39-year-old man presented with a 4-month history of unilateral blepharitis that did not respond to conventional treatment. Punch biopsy confirmed the diagnosis of discoid lupus erythematosus. Unilateral blepharitis as the only presenting sign of discoid lupus erythematosus is uncommon but should be considered in the differential diagnosis in patients with asymmetric blepharitis. PMID- 16714937 TI - Orbital tuberculosis manifesting with enophthalmos and causing a diagnostic dilemma. AB - A 60-year-old woman with no known systemic disease was referred with a hard mass in the left orbit and enophthalmos of two months duration. Differential diagnoses of metastasis from an undetected scirrhous carcinoma and sclerosing nonspecific orbital inflammatory disease were considered and a biopsy was performed. Histopathology demonstrated granulomatous inflammation with fibrosis. Subsequent polymerase chain reaction was positive for Mycobacterium tuberculosis deoxyribonucleic acid. There was no evidence of systemic tuberculosis. The patient was treated with four-drug combination anti-tubercular therapy for 6 months. The mass regressed and there was no local recurrence at two years follow up. Orbital tuberculosis generally manifests with proptosis or osteomyelitis. However, enophthalmos may be caused by the fibrosing variant. Biopsy with histopathologic and microbioloic evaluation is essential to distinguish it from other more common causes of an orbital mass with paradoxical enophthalmos such as metastatic scirrhous carcinoma and sclerosing nonspecific orbital inflammatory disease. PMID- 16714938 TI - Giant epidermoid cyst as an unusual presentation of hypoglobus and ptosis. AB - A 24-year-old man presented with a large right frontal epidural epidermoid cyst that resulted in right hypoglobus and right upper eyelid ptosis. CT and MRI revealed a bone-destroying mass involving the right frontal bone and eroding through the right orbital roof. Complete surgical removal was performed through a frontal craniotomy. After surgery, there was near-complete resolution of the hypoglobus and ptosis. PMID- 16714939 TI - Unrecognized side effect of statin treatment: unilateral blepharoptosis. AB - A 43-year-old man receiving statin monotherapy (10 mg atorvastatin) for hypercholesterolemia had unilateral blepharoptosis as the result of isolated myositis of the levator muscle. Statin-induced myositis in the levator muscle should be considered in the differential diagnosis of acquired unilateral blepharoptosis of unknown cause. PMID- 16714940 TI - Pyogenic granuloma associated with chronic Actinomyces canaliculitis. AB - A case of chronic Actinomyces canaliculitis with associated pyogenic granuloma formation and bloody tears is described. Although Actinomyces is a well-known cause of canaliculitis, the authors are not aware that it has been reported in association with pyogenic granulomas. PMID- 16714941 TI - Maxillary frontal process fracture complicating acute dacryocystorhinostomy. AB - A case of maxillary frontal process fracture complicating an acute endonasal dacryocystorhinostomy is presented. The management of this previously unrecorded complication and the corrective procedure that resulted in both patient and clinician satisfaction is described. PMID- 16714942 TI - Infiltrating orbital astrocytic proliferation associated with congenitally malformed eyes. AB - This report describes the clinical and pathologic findings in two adults with orbital infiltration by astrocytic cells associated with congenitally malformed eyes. Both cases had enlarging orbital masses and underwent complete resection. Histopathology revealed proliferation of astrocytes (fibrillary acidic protein positive) that invaded the orbital tissues. To the best of our knowledge, there are no similar cases in the literature. PMID- 16714943 TI - Malignant hidradenoma of the medial canthus with orbital and intracranial extension. AB - A 70-year-old woman presented with signs and symptoms consistent with nasolacrimal duct obstruction. MRI revealed a medial canthal mass with orbital and intracranial extension. The patient was treated with wide surgical excision, chemotherapy, and radiation. A diagnosis of malignant hidradenoma of the medial canthus was confirmed by histopathology. Despite aggressive therapy, the patient developed metastatic disease and died within 1 year. PMID- 16714944 TI - Oculoplastic approach to treating Barber-Say syndrome. AB - Barber-Say syndrome is a rare dysmorphic disorder with prominent facial involvement. Its primary features include skin, hair, and eyelid abnormalities, the last of which can cause significant ocular symptoms. This report describes a patient with symptomatic corneal exposure secondary to Barber-Say syndrome who benefited from surgical intervention. The patient underwent a bilateral mid-face lift, eyelid repositioning and reconstruction, botulinum toxin application, and acellular dermis grafting. Surgical exploration revealed dysfunctional features of the external and internal eyelid anatomy not previously described with this condition. Improvement was noted in overall facial symmetry and appearance. In addition, the patient noted increased comfort after the resolution of dry eye symptoms and exposure keratopathy. PMID- 16714945 TI - Compressed helium injury to the orbit resulting in pneumocephalus. AB - A 33-year-old woman received a conjunctival laceration after accidental contact with the flexible outlet tip of a tank of compressed helium while filling balloons. The gas discharged during the contact, blowing compressed helium into her right orbit, with intracranial extension. The patient was asymptomatic, except for a transient headache. She was treated with prophylactic antibiotics and observed overnight, then discharged without complication. A literature search reveals that the usual outcome for this mechanism of injury is favorable. PMID- 16714946 TI - Re: "tegaderm transparent dressing (3M) for the treatment of chronic exposure keratopathy". PMID- 16714950 TI - Surgical specimen handling for conjunctival melanoma: implications for tumor thickness determination and sentinel lymph node biopsy. PMID- 16714949 TI - Re: "lateral tarsal strip and quickert sutures for lower eyelid entropion". PMID- 16714951 TI - Immunologic resistance after repeated botulinum toxin type a injections for facial rhytides. PMID- 16714952 TI - Re: "treatment of congenital nasolacrimal duct obstruction with high-pressure irrigation under topical anesthesia". PMID- 16714953 TI - Effect of smoking in graves ophthalmopathy: are pediatric patients exempted? PMID- 16714955 TI - An analysis of pediatric blood cultures in the postpneumococcal conjugate vaccine era in a community hospital emergency department. AB - OBJECTIVE: Blood cultures are commonly included in the evaluation of febrile children younger than 3 years without focal source of infection. Clinicians treat patients with a positive blood culture before final identification of the organism. Their treatment might include reevaluation in the emergency department (ED), additional tests, parenteral antibiotics, and hospital admission even for children who ultimately have false-positive (FP) blood cultures. The advent of pneumococcal conjugate vaccine (PCV) has made occult bacteremia less common, decreasing the likelihood that a positive blood culture result before final organism identification will be a true pathogen. This study will identify the characteristics of patients with FP blood cultures in the post-PCV era. METHODS: Charts were reviewed of all children ages 1 to 36 months with a temperature of at least 38.08 degrees C who had a blood culture obtained in our community hospital ED from January 1997 to January 2005. RESULTS: Bacteria grew in 106 (3.5%) out of 2971 blood cultures. True positives (TPs), defined as true pathogens, had a prevalence of 0.7%, representing 19.8% of positives. FPs, defined as contaminants, occurred in 2.8% of cultures, representing 80.2% of positives. Patients with FP cultures had lower mean white blood cell (WBC) counts (10.51 x 10(9)/L vs. 16.95 x 10(9)/L; P = 0.0001) and lower mean presenting temperatures (38.8 degrees C vs. 39.4 degrees C; P = 0.005). FPs had longer time to positivity (34.6 vs. 17.7 hours; P = 0.001) than TPs. A culture with a Gram stain suggestive of a contaminant, time to positivity greater than 24 hours and a WBC of less than 15 x 10(9)/L had a PPV for an FP of 97%. When analysis was restricted to well appearing children age 2 to 36 months with temperature of more than 39 degrees C without focal source of infection who were discharged from the ED, these three criteria had a PPV for an FP of 100%. In these highly febrile children, the FPs had significantly lower WBCs (9.14 x 10(9)/L vs. 22.84 x 10(9)/L; P = 0.0001) and longer time topositivity (33.4 vs. 19.8 hours; P = 0.007) than TPs. The likelihood of obtaining FP cultures increased after the introduction of PCV from 62.5% to 87.8% odds ratio, 4.3; 95%confidence internal, 1.44-13.38). CONCLUSIONS: In the post-PCV era, the majority of blood culture results will be FPs. FP cultures are predictable in febrile children with WBC counts less than 15.00 x 10(9)/L, time to positivity of more than 24 hours, and a Gram stain result suggestive of a contaminant. Prospective studies applying these criteria to the at-risk population for occult bacteremia are indicated. PMID- 16714956 TI - Maternal and child factors affecting high-volume pediatric emergency department use. AB - OBJECTIVE: To describe maternal and child characteristics associated with high volume nonurgent pediatric emergency department (ED) use. METHODS: We surveyed female caregivers of children aged 1 to 14 years who were brought to the ED at an urban medical center for nonurgent conditions. We analyzed characteristics associated with low-volume (1 ED visit) or high-volume (> or = 2 ED visits) use within the past year. Independent factors included predisposing, reinforcing, and enabling factors. Logistic regression was used to model high-volume ED use. RESULTS: Of the 300 participants (84% response rate), 99 were low-volume users and 201 were high-volume users. Most participants were mothers (94%). In adjusted analyses, high-volume use was associated with caregivers being single [odds ratio (OR), 2.57; 95% confidence interval (CI), 1.13-5.80], having high-volume ED use for themselves (OR, 2.02; 95% CI, 1.08-3.80), seeking care primarily in the pediatric ED for "sick visits" (OR, 2.47; 95% CI, 1.12-5.49), and lack of weekend hours at their child's usual source of care (OR, 2.58; CI, 1.24-5.37). CONCLUSIONS: Marital status, high maternal use of the ED, perception of the ED as the place to seek care for a sick child, and lack of weekend hours for usual source of care are associated with high-volume ED use among children. Given similarities in patterns of ED use among mothers and children, further attention is needed to address clinical practice and system issues that contribute to familial use of EDs. PMID- 16714957 TI - Emergency department visits among pediatric patients for sports-related injury: basic epidemiology and impact of race/ethnicity and insurance status. AB - OBJECTIVES: (1) To characterize the demographics and external causes of pediatric sports injury-related visits (SIRVs) to emergency departments (EDs). (2) To analyze the effect of race/ethnicity and insurance on SIRVs to EDs. METHODS: A stratified random-sample cross-sectional survey of EDs in the National Hospital Ambulatory Medical Care Survey was conducted from 1997-2001; for patients younger than 19 years, we used all visits [n = 33,654; injury-related visits (IRVs) = 13,496, SIRVs = 2990]. We examined both the external cause codes and the actual verbatim text of all IRVs. National estimates of pediatric IRVs were obtained using the assigned patient visit weights in the National Hospital Ambulatory Medical Care Survey databases and SUDAAN 9.1 software (SAS Institute, Inc., Cary, NC). RESULTS: Sports injuries resulted in 2.5 million visits annually, or 23% of ED IRVs. Male sex, older age (6-18 years), and white race/ethnicity are associated with higher rates of SIRVs. Cycling, basketball, playground injuries, and football resulted in the largest numbers of ED SIRVs. Leading diagnoses for SIRVs included fractures and dislocations, sprains and strains, open wounds, and contusions. Hispanic race/ethnicity was associated with lower rates of SIRVs across all insurance types. After controlling for demographic factors and insurance, Hispanic children were less likely to have an SIRV than white children (odds ratio, 0.7; 95% confidence interval, 0.6-0.9). CONCLUSIONS: Sports and recreation are the leading causes of pediatric ED IRVs. Hispanic children, regardless of insurance status, had lower rates of SIRVs than white children, which helps explain the lower rate of nonfatal IRVs to EDs among Hispanic youth. PMID- 16714958 TI - The interrater reliability of a validated bronchiolitis severity assessment tool. AB - BACKGROUND: We previously constructed and tested a bronchiolitis severity assessment tool in 2 independent hospitals. The model uses age, work of breathing, dehydration and tachycardia to successfully predict disease severity. OBJECTIVE: To prospectively measure the interrater reliability of a bronchiolitis severity assessment tool and of its component variables. DESIGN: Prospective observational survey. SETTING: A county teaching hospital emergency department serving a mixed urban and rural population with an emergency medicine residency program in 2-3-4 format. SUBJECTS: Thirty-two physicians evaluated a convenience sample of children aged less than 18 months presenting to the emergency department with a clinical diagnosis of bronchiolitis during a single season. METHODS: Two physicians independently examined each patient. Each physician completed a physical examination template that included the variables used in the severity assessment tool. Interrater agreement was measured for the variables work of breathing and dehydration and for the tool as a whole using a weighted kappa statistic. RESULTS: One hundred and forty-six cases were enrolled. Twenty five were dropped for incomplete data collection. The actual weighted agreement on overall classification was 92%; expected, 73%, kappa = 0.676; P < 0.0001. The actual weighted agreement for dehydration was at 95%; expected, 92%, kappa = 0.305; P = 0.0001. The agreement for work of breathing was 95%; expected, 86%; kappa = 0.611; P < 0.0001. The overall model showed better interrater reliability than its individual components. CONCLUSIONS: Overall interrater reliability for this bronchiolitis severity assessment tool is substantial. PMID- 16714959 TI - Too long in the tooth: a descriptive study of adults presenting to a pediatric emergency department. AB - OBJECTIVE: To describe the demographics, presenting complaints, diagnoses, and disposition of adults presenting to an Australian pediatric emergency department. It was hypothesized that most patients would be younger than 20 years, with low acuity complaints predominantly related to minor injuries resulting in a low admission rate. METHODS: A prospective cohort of new adult patients (age, > or = 16 years) were recruited. Baseline data was recorded for all eligible patients. Consenting patients had provisional diagnoses recorded along with disposition decisions. Patients' records were reviewed at the referral destination to establish the final diagnoses and the admission rate. RESULTS: Forty-eight patients (0.6% of all presentations) presented during the study period. The median age was 17.5 years (interquartile range, 16.3-26.1 years). Thirty-four patients (70.8%) were younger than 20 years (P = 0.003). Seven (14.6%) patients had an Australasian Triage Scale score of 2, and 19 (39.6%) had a score of 3. Injury-related complaints formed the largest proportion of presentations (mean, 37.5%; range, 23.8%-51.2%). Thirty-one patients were enrolled in the study. Twenty (mean, 48.8%; range, 33.5%-64.1%) of the provisional diagnoses in this group were injury-related. Twenty-nine patients (mean, 93.5%; range, 84.9%-100%) were advised to seek further medical attention. Injuries comprised the largest group of final diagnoses (mean, 55.2%; range, 37.1%-73.3%). Five patients (mean, 16.1%; range, 3.2%-29%) were admitted. CONCLUSIONS: Adult presentations were not infrequent. Twenty patients (41.7%) were within 1 year of the age cutoff at Princess Margaret Hospital. Although 45.8% of presentations had a low acuity, the majority of patients had Australasian Triage Scale scores of 2 or 3. The presentations were heterogeneous in nature, with injuries accounting for a large proportion. Most patients were referred for further care, but a minority were not compliant with follow-up instructions. There was a low admission rate. PMID- 16714960 TI - Emergency visits for childhood poisoning: a 2-year prospective multicenter survey in Spain. AB - OBJECTIVES: To describe the characteristics of childhood poisoning leading to consultation to 17 pediatric emergency departments in Spain. METHODS: During a 2 year period (January 2001 to December 2002), accompanying people of 2157 children with acute intoxication who visited consecutively at the emergency room were prospectively surveyed. RESULTS: Childhood poisoning accounted for 0.28% of all emergency visits during the study period. The median (interquartile range, 25th 75th percentile) age was 24 months (22-60 months); 67% of children were younger than 4 years. Drug ingestion was involved in 54.7% of cases (paracetamol was the most frequent drug), domestic products in 28.9%, alcohol in 5.9%, carbon monoxide in 4.5%, and illicit drugs in 1.5%. A total of 61.3% of patients were admitted within 1 hour after exposure to the toxic substance, and 10.3% had been already treated before arrival; 29.1% of patients were referred for clinical manifestations which were mostly neurological symptoms. Laboratory tests and other investigations were performed in 40.7% of cases. Gastrointestinal decontamination was used in 51.7% of patients, with activated charcoal in 32.3%. Treatment varied significantly according to the individual hospitals. A total of 83.3% of patients were treated as outpatients, 15.2% were hospitalized, and 1.5% were admitted to the intensive care unit. One 11-month-old boy with carbon monoxide intoxication died. Six patients had permanent sequelae (esophageal stenosis in 5 and partial blindness in 1). CONCLUSIONS: Young children who accidentally ingested drugs and, less frequently, domestic products accounted for most cases of intoxication who presented at the pediatric emergency department. PMID- 16714961 TI - Brief focal ultrasound with topical anesthetic decreases the pain of intravenous placement in children. AB - OBJECTIVE: To determine whether brief, focal pretreatment of children's skin with low-frequency ultrasound followed by a 5-minute application of a 4% lidocaine topical anesthetic decreases the pain of intravenous (IV) catheter placement. METHODS: A randomized, double-blind, placebo-controlled trial enrolling children 8 to 18 years of age undergoing IV placement as part of their care in a pediatric emergency department. Thirty-eight children received pretreatment followed by 5 minute application of a topical anesthetic. Thirty-nine children received pretreatment followed by 5-minute application of a placebo cream. Children and parents rated the pain associated with IV placement using the visual analog scale (VAS). RESULTS: There were no significant differences in demographics between the 2 groups. Children in the treatment group had significantly lower VAS scores (mean, 2.29) than children in the control group (mean, 3.23) (P = 0.023; 95% confidence interval of the mean difference, -1.87 to -0.02). The parents' VAS scores of the child's pain were also lower for the treatment group (mean, 2.47 vs 3.39; P = 0.038; confidence interval, -1.97 to 0.11). CONCLUSIONS: Visual analog scale scores measuring children's pain and parents' perception of the child's pain were lower in those who were pretreated with brief focal ultrasound and 5 minutes of 4% lidocaine cream when compared with those pretreated with ultrasound and placebo. PMID- 16714962 TI - Megacolon: constipation or volvulus? AB - Pediatric abdominal complaints are common presentations in emergency departments, and emergency physicians are challenged every day with a vast array of gastrointestinal disorders. Differentiating the child with a benign abdominal process from the child with a more serious condition can be difficult. We report a case of massive dilatation of the colon due to fecal impaction. Means for distinguishing constipation from alternative diagnoses are discussed. PMID- 16714963 TI - A teenage girl with extreme tachypnea. AB - Dyspnea and tachypnea are common presenting complaints in an emergency department. Respiratory and cardiac causes are the most common etiology. Functional respiratory disorders should be considered in patients with atypical presentation to avoid extensive investigations. We describe an adolescent who presented to our emergency department with extreme tachypnea. PMID- 16714964 TI - Intentional avulsion fracture of the second cervical vertebra in a hypotonic child. AB - Children uncommonly present with cervical fractures due to high-impact injuries. A 4-year-old child with developmental delay and chronic hypotonia presented to the hospital with acute onset of irritability and transient difficulty in walking. Lateral neck x-rays showed avulsion fracture of the second cervical vertebra. Neck magnetic resonance imaging study showed injury to the interspinal ligaments between the first and second cervical vertebrae in addition to confirming the acute avulsion fracture of C-2. The child was injured after he was thrown onto a bed from a distance of 1 to 2 ft according to his babysitter's admission. This is the first reported pediatric case presenting this type of inflicted fracture in a hypotonic child, although hypotonia may not have played a causal role. PMID- 16714965 TI - Turpentine oil inhalation leading to lung necrosis and empyema in a toddler. AB - Household poisons can cause serious damage to the health of children. Turpentine oil, used chiefly as a solvent in paints, varnishes, and waxes; is often placed within easy reach of children during polishing or painting work. It is capable of causing serious toxicity, whether ingested or inhaled. Pulmonary aspiration can lead to lung parenchymal damage and turpentine-associated pneumonia. We report a case of exploratory ingestion and inhalation of turpentine oil in an 18-month-old boy, leading to extensive lung parenchymal damage, formation of multiple abscess cavities, and necrosis treated with decortication and segmental resection. PMID- 16714966 TI - Lung injury from "liquid ecstasy": a role for coagulation activation? AB - We describe our second clinical observation of pulmonary injury after a "liquid ecstasy" ingestion/inhalation by a 3-year-old girl. Apart from the deep coma state, a markedly asymmetric pulmonary compromise was recorded. A transient coagulation activation was detected, possibly triggered off by the toxic effect on the lung alveolar-capillary membrane. PMID- 16714967 TI - Healthy children with invasive community-acquired methicillin-resistant Staphylococcus aureus infections. AB - Reports of community-acquired methicillin-resistant Staphylococcus aureus (CA MRSA) infections in the pediatric community have exploded during the past decade. These infections typically result in mild skin and soft tissue infections that can be managed simply with oral antimicrobials. Recently, there have been reports of invasive CA-MRSA infecting children without risk factors, with isolated cases of life-threatening disease. We report 2 atypical cases of invasive CA-MRSA infecting previously healthy children. PMID- 16714968 TI - Acute hip pain in the nonambulatory infant: Salter-Harris type I fracture in the capital femoral epiphysis without a traumatic history. AB - An otherwise healthy 11-month-old girl was brought to the hospital after her parents noted the acute onset of right hip pain and refusal to bear weight. No abnormalities were seen in the initial radiographs, laboratory values were within reference range, and noninvasive workup was negative for septic arthritis. The parents reported a recent minor fall from a standing position, but stated that the child seemed to return to normal without pain after a few minutes of crying. A hemarthrosis without purulence was found upon joint aspiration, and the patient improved significantly after administration of anti-inflammatory medication. Follow-up radiographs 13 days after initial presentation showed an extremely rare Salter-Harris type I proximal physeal fracture well into the healing process. PMID- 16714969 TI - Management of esophageal coins. PMID- 16714971 TI - Pediatric transport team: Intermountain Life Flight. AB - The transport of the pediatric patient presents unique challenges. To assure the children receive the care they need during transport, the appropriate specialty team should be used. This article presents the role of a pediatric specialty team in patient transport and offers an example of a long-standing pediatric transport team. PMID- 16714972 TI - Child and adolescent psychiatric emergencies in nonsuicide-specific samples: the state of the research literature. AB - OBJECTIVE: Since the 1980s, there has been an increase in the frequency with which children and adolescents with psychiatric symptoms present to emergency departments (ED). Recent trend data from the National Hospital Ambulatory Medical Care Survey database suggest that the increase in ED use for mental health concerns is not driven by a rise in the frequency of medically serious suicide attempts but, rather, nonurgent diagnoses. The current paper is a systematic review of the existing research literature on child and adolescent psychiatric emergencies among nonsuicidal samples to assess the type and quality of the data reported. This is a first step in developing a profile of which children, in the absence of medically serious suicide attempt, seek psychiatric care in EDs. METHODS: Literature searches were conducted through PsycINFO, MEDLINE, and PubMed databases for the years 1990 through 2004. Twelve studies were identified as meeting inclusion criteria and were reviewed for general study information (ie, bibliography); quality indicators; study characteristics (ie, data source, location of study, method of subject recruitment, and study inclusion/exclusion criteria); subject characteristics, and outcome measures and results (specific outcome assessed, types of data analyses). RESULTS: Most of the studies were cross-sectional, retrospective chart reviews, with one data source. Only one study reported on all variables of interest, and there was no consistency in data collection or reporting across studies. CONCLUSIONS: Although there has been a documented increase in children and adolescents seeking care for psychiatric emergencies in EDs, a clear and basic understanding of who these patients are cannot be determined from existing published research reports. Significant inconsistencies in data collection and reporting make recommendations for treatment planning and service delivery challenging. A research agenda in the field of child and adolescent psychiatric emergencies is proposed. PMID- 16714973 TI - ECGs in the ED. PMID- 16714974 TI - Pediatric emergency medicine: legal briefs. PMID- 16714975 TI - Head trauma and hospital admission. PMID- 16714977 TI - Hyperbaric oxygen therapy for purpura fulminans-comment. PMID- 16714978 TI - Aspiration hazards of cosmetics containing silicone derivatives. PMID- 16714980 TI - Future shock? PMID- 16714981 TI - Hypotension and mortality in septic shock: the "golden hour". PMID- 16714982 TI - The pulmonary artery catheter and critical care: the cart is before the horse. PMID- 16714983 TI - Blood transfusion in burns: benefit or risk? PMID- 16714985 TI - Achieving optimal antiarrhythmic therapy in advanced cardiac life support. PMID- 16714984 TI - Blood product transfusion in association with coronary artery bypass grafting: proceed with caution. PMID- 16714987 TI - Rib fractures, pneumonia, and mortality. PMID- 16714986 TI - Is fast-track intensive care unit management still on the express track? PMID- 16714988 TI - Transmitting and absorbing new information on the early identification of sepsis patients: the partial thromboplastin time biphasic waveform. PMID- 16714989 TI - Pharmacokinetic and pharmacodynamic research in the intensive care unit: an unmet need. PMID- 16714990 TI - Permissive hypercapnia: does a high PaCO2 level require high sedative doses? PMID- 16714991 TI - Do intensive care unit patients have intensive care unit physicians? Unfortunately not. PMID- 16714992 TI - Family satisfaction from clinician statements or patient-provider concordance? PMID- 16714993 TI - End-of-life variations around the world: can we improve our caring? PMID- 16714994 TI - Managing pain and agitation in the critically ill--are we there yet? PMID- 16714995 TI - Medical emergency teams: are improved outcomes really like day and night? PMID- 16714996 TI - Beta-2-agonist treatment as a potential therapy for acute inhalational lung injury. PMID- 16714997 TI - Do coagulation abnormalities contribute to sepsis associated organ failure? PMID- 16714998 TI - To breathe or not to breathe: is spontaneous ventilation the answer for acute lung injury? PMID- 16714999 TI - Sepsis and stretch: synergistic effects on alveolar epithelial cell death? PMID- 16715000 TI - L-arginine and vasopressor agents: when antagonists have unexpected synergistic effects. PMID- 16715001 TI - Resuscitation with hypertonic saline in burn shock and sepsis. PMID- 16715002 TI - Brain tissue oxygen monitoring: a measure of supply and demand. PMID- 16715003 TI - Critical illness: a view from the kidney. PMID- 16715005 TI - Precise use of medical terminology. PMID- 16715007 TI - Noninvasive ventilation and "do not intubate". PMID- 16715008 TI - Glycemic control needs a standard reference point. PMID- 16715012 TI - Preoxygenation remains essential before emergency tracheal intubation. PMID- 16715011 TI - Exploring the role of etomidate in septic shock and acute respiratory distress syndrome. PMID- 16715013 TI - Errors in table, not "diametrically opposite result"! PMID- 16715016 TI - Assessment of the validity of masking in electrodiagnostic research. AB - OBJECTIVES: Performed by thousands of specialists and researched for >60 yrs, electrodiagnostic testing rates poorly in evidence-based guidelines in part because there are no valid masked research studies. This study assesses whether the electrodiagnostic examination can be adequately masked. DESIGN: Prospective, controlled, masked trial. A total of 150 persons aged 55-80 yrs, thought clinically to be asymptomatic (n = 32), with low back pain (n = 46), or with spinal stenosis (n = 63), underwent needle electromyography and nerve conduction studies by specialists in physical medicine and rehabilitation who were not permitted to ask any clinical information or to perform a formal examination. Examiners rated potential bias and recorded clues that might bias the examination. RESULTS: Examiners noted potential for bias in conversation in 17 and due to pain behavior in one. Half of these potential unmaskings pointed away from the correct clinical diagnosis, leaving potential for bias in <6% of the subjects. The clinical diagnosis had no relationship to electrodiagnostician rating of pain behavior in response to testing or repositioning, to limb atrophy, and to arthritis (P > 0.05 all instances). CONCLUSIONS: The electrodiagnostic examination can be successfully masked. This information will help future research in electrodiagnostics meet the criteria of evidence-based medicine. PMID- 16715017 TI - Outcomes in total knee arthroplasty patients after inpatient rehabilitation: influence of age and gender. AB - OBJECTIVES: To examine age and gender effects on outcomes after inpatient rehabilitation in primary total knee arthroplasty (TKA) and revision TKA patients. DESIGN: This retrospective, comparative study was conducted in a 50 bed, university-affiliated rehabilitation hospital. Patients included primary TKA patients (n = 286) and revision TKA patients (n = 138) placed into one of three age brackets: <60 yrs, 60-70 yrs, or >70 yrs. Measurements included length of stay (LOS), FIM scores, hospital charges, and discharge disposition location. RESULTS: FIM scores improved from admission to discharge in both primary and revision patients (P = 0.015). LOS was shorter in primary than revision patients (8.3 vs. 10.4 days, P = 0.001), and the LOS was longest in patients >70 yrs compared with those <60 or 60-70 yrs (10.6 vs. 8.7 and 8.8 days, respectively; P = 0.004). FIM efficiency was greater in primary than in revision TKA patients (3.68 vs. 2.77 points/day, P = 0.001), and greater for men than women in each age bracket regardless of TKA type (3.68 vs. 2.78 points/day, P = 0.001). Total hospital charges were lower for men than women for both TKA types ($9,656 +/- 823 vs. $11,544 +/- 1,359; P = 0.015), and were highest in patients >70 yrs of age (P = 0.015). CONCLUSIONS: Primary and revision TKA patients make improvements in functional independence during inpatient rehabilitation. Although FIM gains were similar among age and gender groupings, FIM efficiency is lowest in women. Despite similar daily hospital costs among the groups, slower progress with functional gains (decreased FIM efficiency) increases the LOS and total hospital charges, especially in older women. PMID- 16715018 TI - Evaluating treatment effectiveness: benchmarks for rehabilitation after partial meniscectomy knee arthroscopy. AB - OBJECTIVE: The purpose of this study was to give a detailed description of recovery benchmarks that occur in patients whose therapy after partial meniscectomy knee arthroscopy consists of a home program of exercise. These benchmarks can be used as a basis for clinicians to compare improvements to individual patients who receive supervised care. DESIGN: Thirty-nine patients (five females, mean age = 41) who underwent an uncomplicated arthroscopic partial meniscectomy were included. Test sessions occurred at 5 and 50 days after surgery. Outcome measures included: 1) Hughston Clinic knee self-assessment questionnaire; 2) EQ-5D Tariff for assessment of quality of life; 3) number of days taken to return to work after surgery; 4) knee passive range of motion; and 5) knee swelling assessed by evaluation of knee circumference. Stepwise regression analysis was used to evaluate factors that might have influenced the amount of pre- to posttest change in the outcome measures (the benchmarks) during the first 7 wks after surgery. The factors used in this analysis were: 1) age, 2) body mass index, 3) period from injury to surgery, and 4) the baseline value of the variable to be examined (except for return to work, where we used a score estimating the challenge to the knee offered by work). RESULTS: None of the factors considered (age, body mass index, period from injury to surgery, stressfulness of the work on the knee) affected the number of days taken to return to work. Baseline scores affected change in all the other outcomes, and knee girth change was also affected by body mass index. Regression equations are presented where suitable for the benchmarks presented. CONCLUSIONS: Quick recovery occurs in these patients when only a home exercise program is given. This paper highlights the utility of using historical control group data instead of test-retest analysis of measurement error in evaluating patients whose recovery with a home exercise program is rapid. Of the variables analyzed in this study, quality of life and knee self-assessment changes offer the most useful benchmarks for evaluating treatment effectiveness. PMID- 16715019 TI - Reliability of a new instrument for measuring maximum rising strength. AB - OBJECTIVE: To assess the intrarater test-retest reliability of a newly developed instrument for measuring maximum rising strength (MRS) in comparison with that of maximum knee extension strength (KES). DESIGN: Three replications of isometric MRS and KES were measured on three test occasions separated by 1 wk in 97 healthy men and women aged 23-90. MRS was measured in seated subjects using a dynamometer fixed on the ground and connected by an adjustable nonelastic cord to a padded belt. RESULTS: The intraclass coefficients (ICCs) of MRS were 0.92 and 0.93 at 60 and 70 degrees of knee flexion, respectively, and 0.98 when the mean of three highest peak values of MRS at 60 degrees and the three highest peak values at 70 degrees knee flexion on each session were considered vs. 0.95 for KES. A significant increase in MRS and KES strength was found between the first and the third sessions (+7 to 8%)(P < 0.001). No side effects of strength measurements were noticed. CONCLUSION: Despite a significant learning effect, MRS measurement is reliable in men and women of a wide range of ages. Whether this instrument will be useful for targeting individuals with early sit-to-stand difficulties requires further investigation. PMID- 16715020 TI - Changes in posture and perceived exertion in adolescents wearing backpacks with and without abdominal supports. AB - OBJECTIVE: The goal was to examine whether backpacks with an abdominal support device improve posture and decrease exertion while walking among adolescents. DESIGN: Double-blinded trial, with 20 subjects, performed at a University Spine Center. On day 1, adolescents walked on a treadmill for 5 mins with photographic measurement of posture and responded to the Borg scale of perceived exertion before and after ambulation. On day 2, they repeated the trial four more times wearing a backpack randomly loaded with either 10% or 20% of body weight and with or without the Back Balancer abdominal support. Postural measures were recorded by a blinded observer. RESULTS: Postural changes (forward lean) wearing a backpack with abdominal support were significantly lower while carrying both 10% and 20% of body weight (P = 0.024 and P = 0.008, respectively) vs. no abdominal support. In addition, perceived exertion at 10% and 20% of body weight was significantly greater without abdominal support (P = 0.042 and P = 0.018, respectively). CONCLUSIONS: Forward lean and perceived exertion while wearing backpacks in adolescents can be significantly decreased with abdominal support. This may increase comfort and decrease complications, such as back pain, that may arise from backpack use. PMID- 16715021 TI - Effect of manual lymph drainage on the course of serum levels of muscle enzymes after treadmill exercise. AB - OBJECTIVE: Improving muscular recovery after exercise is an important topic in sports medicine. The aim of the present study was to evaluate the effect of manual lymph drainage on the course of serum levels of muscle enzymes after an extended treadmill exercise. DESIGN: Fourteen recreational athletes (seven women, seven men) were included in the study. The participants underwent a graded exercise test on a treadmill ergometer to determine the individual anaerobic threshold (IAT). Seven days after the graded exercise test, all subjects performed 30 mins of treadmill exercise at an intensity equivalent to IAT. The subjects were randomized into two groups of seven persons. One group was treated with manual lymph drainage (ML), whereas the control group (CG) received no treatment after the endurance exercise at IAT level. RESULTS: After an increase immediately after exercise, a fast decrease in lactate dehydrogenase (LDH) and in aspartate aminotransferase (AST) concentration was observed, with significantly lower values for LDH after 48 hrs in the subjects having received lymph drainage treatment. The course of creatine kinase (CK) levels was comparable, but did not reach significance. CONCLUSION: Manual lymph drainage after treadmill exercise was associated with a faster decrease in serum levels of muscle enzymes. This may indicate improved regenerative processes related to structural damage of muscle cell integrity. PMID- 16715022 TI - Falls sustained during inpatient rehabilitation after lower limb amputation: prevalence and predictors. AB - OBJECTIVE: The purpose of this study is to identify risk factors for falling and fall-related injury among a group of inpatients undergoing rehabilitation after major lower limb amputation. DESIGN: Retrospective cohort. RESULTS: Out of 1267 patients, 260 (20.5%) fell at least once. There were a total of 374 falls, 67 (17.9%) of which resulted in one or more injuries. Adjusted odds ratios (OR) and 95% confidence intervals (CI) were calculated for factors significantly associated with falling, including age of > or =71 yrs (OR = 1.40, 95% CI = 1.02 1.89), lengths of stay of 22-35 days (OR = 2.97, 95% CI = 1.14-7.72) or >5 wks (OR = 6.07, 95% CI = 2.34-15.71), four or more comorbidities (OR = 1.93, 95% CI = 1.09-3.41), cognitive impairment (OR = 1.68, 95% CI = 1.02-2.78), two or more as needed medications (OR = 1.81, 95% CI = 1.02-3.21), benzodiazepines (OR = 2.22, 95% CI = 1.24-3.96), and opiates (OR = 5.76, 95% CI = 3.29-10.09). Factors significantly associated with fall-related injuries included bilateral amputation (OR = 3.68, 95% CI = 1.49-9.05) and falls during the day shift (OR = 2.63, 95% CI = 1.24-5.57). CONCLUSIONS: One in five patients with lower limb amputation will likely experience at least one fall during inpatient rehabilitation, with 18% sustaining an injury. Ongoing research is required to develop appropriate intervention strategies to ameliorate the risk of falling during inpatient rehabilitation. PMID- 16715024 TI - Pharmacotherapy for neurogenic detrusor overactivity. AB - Chancellor MB, Anderson RU, Boone TB: Pharmacotherapy for neurogenic detrusor overactivity. Am J Phys Med Rehabil 2006;85:536-545. Patients with neurogenic detrusor overactivity are a heterogeneous group with voiding dysfunction secondary to neurologic injury or disease. The neurogenic detrusor overactivity syndrome, which may include urinary frequency, urgency, and incontinence, frequently contributes to a loss of independence, or even institutionalization. Urodynamic assessment provides the best method of quantifying and classifying neurogenic detrusor overactivity dysfunction in patients with primary diagnoses as diverse as Parkinson's disease, cerebral palsy, multiple sclerosis, spinal cord injury, and spina bifida. For many patients, management of urinary symptoms includes pharmacotherapy with an anticholinergic agent. Several novel approaches to managing neurogenic detrusor overactivity, including intravesical instillation of anticholinergic agents, vanilloids, and neurotoxins, are being investigated. For most patients, however, flexible dosing with an anticholinergic agent, with clean intermittent catheterization when indicated, has been shown to reduce the risks of urologic complications, improve levels of continence, and enhance patient quality of life in both children and adults. PMID- 16715026 TI - Femoroacetabular impingement: an overlooked cause of hip pain. AB - Femoroacetabular impingement is a cause of hip pain arising from abnormal contact between the proximal femur and the acetabulum during terminal hip motion. This is caused by a structural abnormality at the femoral head/neck junction or the acetabulum. The problem usually presents in young adults and can often go undiagnosed for years. Failure to identify and properly restore the biomechanics has been hypothesized to lead to the early onset of hip osteoarthritis; thus, early recognition is important. In this case study, we report on a 31-yr-old patient with many years of bilateral hip pain who underwent successful treatment of bilateral femoroacetabular impingement. PMID- 16715027 TI - The absurdity of smoking (either tobacco or marijuana). PMID- 16715028 TI - Myotonic dystrophy. PMID- 16715029 TI - How will we know patients are safer? An organization-wide approach to measuring and improving safety. AB - OBJECTIVE: Our institution, like many, is struggling to develop measures that answer the question, How do we know we are safer? Our objectives are to present a framework to evaluate performance in patient safety and describe how we applied this model in intensive care units. DESIGN: We focus on measures of safety rather than broader measures of quality. The measures will allow health care organizations to evaluate whether they are safer now than in the past by answering the following questions: How often do we harm patients? How often do patients receive the appropriate interventions? How do we know we learned from defects? How well have we created a culture of safety? The first two measures are rate based, whereas the latter two are qualitative. To improve care within institutions, caregivers must be engaged, must participate in the selection and development of measures, and must receive feedback regarding their performance. The following attributes should be considered when evaluating potential safety measures: Measures must be important to the organization, must be valid (represent what they intend to measure), must be reliable (produce similar results when used repeatedly), must be feasible (affordable to collect data), must be usable for the people expected to employ the data to improve safety, and must have universal applicability within the entire institution. SETTING: Health care institutions. RESULTS: Health care currently lacks a robust safety score card. We developed four aggregate measures of patient safety and present how we applied them to intensive care units in an academic medical center. The same measures are being applied to nearly 200 intensive care units as part of ongoing collaborative projects. The measures include how often do we harm patients, how often do we do what we should (i.e., use evidence-based medicine), how do we know we learned from mistakes, and how well do we improve culture. Measures collected by different departments can then be aggregated to provide a hospital level safety score card. CONCLUSION: The science of measuring patient safety is immature. This article is a starting point for developing feasible and scientifically sound approaches to measure safety within an institution. Institutions will need to find a balance between measures that are scientifically sound, affordable, usable, and easily applied across the institution. PMID- 16715030 TI - Impact of patient volume on the mortality rate of adult intensive care unit patients. AB - OBJECTIVE: Expert task forces have proposed that adult critical care medicine services should be regionalized in order to improve outcomes. However, it is currently unknown if high intensive care unit (ICU) patient volumes are associated with reduced mortality rate. The objective was to investigate whether high-volume ICUs have better mortality outcomes than low-volume ICUs. DESIGN: Retrospective cohort study analyzing the association between ICU volume and in hospital mortality using Project IMPACT (a clinical outcomes database created by the Society of Critical Care Medicine). PATIENTS: The analyses were based on 70,757 patients admitted to 92 ICUs between 2001 and 2003. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The main outcome measure was in-hospital mortality. Hierarchical logistic regression modeling was used to examine the volume-outcome association. The median (interquartile range) ICU volume was 827 (631-1,234) patient admissions per year. The overall mortality rate was 14.6%. After controlling for patient risk factors and ICU characteristics, and clustering, there was evidence that patients admitted to high-volume ICUs had improved outcomes (p = .025). However, this mortality benefit was seen only in high-risk patients treated at ICUs treating high volumes of high-risk patients. CONCLUSIONS: There is evidence that high ICU patient volumes are associated with lower mortality rates in high-risk critically ill adults. PMID- 16715031 TI - Procalcitonin as a diagnostic test for sepsis in critically ill adults and after surgery or trauma: a systematic review and meta-analysis. AB - OBJECTIVE: To quantify the accuracy of serum procalcitonin as a diagnostic test for sepsis, severe sepsis, or septic shock in adults in intensive care units or after surgery or trauma, alone and compared with C-reactive protein. To draw and compare the summary receiver operating characteristics curves for procalcitonin and C-reactive protein from the literature. DATA SOURCE: MEDLINE (keywords: procalcitonin, intensive care, sepsis, postoperative sepsis, trauma); screening of the literature. STUDY SELECTION: Meta-analysis of all 49 published studies in medical, surgical, or polyvalent intensive care units or postoperative wards. Children, medical patients, and immunocompromised patients were excluded. DATA EXTRACTION: Thirty-three studies fulfilled inclusion criteria (3,943 patients, 1,828 males, 922 females; mean age: 56.1 yrs; 1,825 patients with sepsis, severe sepsis, or septic shock; 1,545 with only systemic inflammatory response syndrome); eight studies could not be analyzed statistically. Global mortality rate was 29.3%. DATA SYNTHESIS: Global odds ratios for diagnosis of infection complicated by systemic inflammation were 15.7 for the 25 studies (2,966 patients) using procalcitonin (95% confidence interval, 9.1-27.1) and 5.4 for the 15 studies (1,322 patients) using C-reactive protein (95% confidence interval, 3.2-9.2). The summary receiver operating characteristics curve for procalcitonin was better than for C-reactive protein. In the 15 studies using both markers, the Q* value (intersection of summary receiver operating characteristics curve with the diagonal line where sensitivity equals specificity) was significantly higher for procalcitonin than for C-reactive protein (0.78 vs. 0.71, p = .02), the former test showing better accuracy. CONCLUSIONS: Procalcitonin represents a good biological diagnostic marker for sepsis, severe sepsis, or septic shock, difficult diagnoses in critically ill patients. Procalcitonin is superior to C reactive protein. Procalcitonin should be included in diagnostic guidelines for sepsis and in clinical practice in intensive care units. PMID- 16715032 TI - Activated protein C therapy in a rat heat stroke model. AB - OBJECTIVE: To evaluate the therapeutic effects of activated protein C in an animal model of heat stroke. DESIGN: Laboratory investigation. SETTING: Chi-Mei Medical Center research laboratory. SUBJECTS: Male Sprague-Dawley rats weighing 252-304 g. INTERVENTIONS: Anesthetized animals were subjected to heat stress (40 degrees C) to induce heat stroke. A bolus injection of normal saline or recombinant human activated protein C (drotrecogin alfa, activated) was conducted via femoral catheters immediately after the onset of heat stroke. Blood sampling was done before initiation of heat stress and 0 and 40 mins after the onset of heat stroke. MEASUREMENTS AND MAIN RESULTS: When the vehicle-treated rats underwent heat exposure, their survival time values were found to be 56-64 mins (n = 16). Resuscitation with activated protein C significantly and dose dependently improved survival during heat stroke (108-246 mins for doses of 0.5 20 mg of activated protein C per kilogram of body weight) (n = 32). All heat stressed animals displayed systemic inflammation and activated coagulation, evidenced by increased tumor necrosis factor-alpha, prothrombin time, activated partial thromboplastin time, and D-dimer and decreased platelet count and protein C. Biochemical markers evidenced by cellular ischemia and injury/dysfunction included increased plasma levels of blood urea nitrogen, creatinine, glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, and alkaline phosphatase; increased striatal levels of glutamate, glycerol, and lactate/pyruvate ratio; and decreased striatal levels of partial pressure of oxygen and local cerebral blood flow, which were all observed during heat stroke. These heat stroke reactions were all significantly suppressed by resuscitation with activated protein C but not vehicle solution. CONCLUSIONS: The results indicate that systemic delivery of human recombinant activated protein C at the time point of onset of heat stroke may improve survival by ameliorating systemic inflammation, hypercoagulable state, and tissue ischemia and injury in multiple organs. PMID- 16715033 TI - A curriculum in critical care medicine for maternal-fetal medicine fellows. AB - OBJECTIVE: Critical care education in maternal-fetal medicine has typically been limited or has been driven entirely by clinical cases. A pilot program was created to systematize critical care training within the context of an existing maternal-fetal medicine fellowship. DESIGN: The program included a lecture series, a Web-based learning module, specific scenarios played out in a human simulator lab, and a clinical intensive care unit rotation. Fellows' knowledge was assessed by pre- and posttest. RESULTS: Objective knowledge was demonstrated to increase after 1 yr of training. Fellows' impression of the curriculum was positive. CONCLUSION: More structured critical care medicine training is feasible within a maternal-fetal medicine fellowship. A multimodal curriculum to teach critical care within a maternal-fetal medicine fellowship appears promising and could be implemented elsewhere. Interest in this area should be cultivated. PMID- 16715034 TI - Effects of drotrecogin alfa activated on microcirculatory alterations in patients with severe sepsis. AB - OBJECTIVE: Microvascular alterations may play an important role in the development of sepsis-induced organ dysfunction. Drotrecogin alfa activated (DAA) improves outcome in patients with severe sepsis, but its precise mechanism of action is not entirely defined. We investigated whether DAA can influence microcirculatory alterations in patients with severe sepsis. DESIGN: Prospective, nonrandomized study. SETTING: A 31-bed, medico-surgical intensive care unit of a university hospital. PATIENTS: Forty adult patients with severe sepsis who met the EU criteria for DAA administration. INTERVENTIONS: Twenty patients received the drug (DAA) and 20 had a contraindication to DAA administration (control). MEASUREMENTS AND MAIN RESULTS: An orthogonal polarization spectral imaging technique was used to visualize the sublingual microcirculation. In all patients, measurements were obtained at baseline, 4 hrs later, and then every 24 hrs for up to 7 days. In patients receiving DAA, measurements were also obtained just before and 4 hrs after the end of DAA infusion. The two groups were well matched for severity of disease, number of failing organs, and the degree of microvascular alterations at baseline. The proportion of perfused capillaries increased in the DAA treated patients already at 4 hrs (from 64% [51-80%] to 84% [71-88%], p < .01) but not in the control group (from 67% [59-76%] to 68% [61-71%], p = not significant). Microvascular perfusion decreased transiently at the end of DAA infusion. The improvement in microvascular blood flow was associated with a more rapid resolution of hyperlactatemia. CONCLUSIONS: DAA administration rapidly improves sepsis-induced microvascular alterations, whereas its cessation is associated with a transient deterioration. PMID- 16715035 TI - From evidence to clinical practice: effective implementation of therapeutic hypothermia to improve patient outcome after cardiac arrest. AB - OBJECTIVES: Therapeutic hypothermia has been recommended for postcardiac arrest coma due to ventricular fibrillation. However, no studies have evaluated whether therapeutic hypothermia could be effectively implemented in intensive care practice and whether it would improve the outcome of all comatose patients with cardiac arrest, including those with shock or with cardiac arrest due to nonventricular fibrillation rhythms. DESIGN: Retrospective study. SETTING: Fourteen-bed medical intensive care unit in a university hospital. PATIENTS: Patients were 109 comatose patients with out-of-hospital cardiac arrest due to ventricular fibrillation and nonventricular fibrillation rhythms (asystole/pulseless electrical activity). INTERVENTIONS: We analyzed 55 consecutive patients (June 2002 to December 2004) treated with therapeutic hypothermia (to a central target temperature of 33 degrees C, using external cooling). Fifty-four consecutive patients (June 1999 to May 2002) treated with standard resuscitation served as controls. Efficacy, safety, and outcome at hospital discharge were assessed. Good outcome was defined as Glasgow-Pittsburgh Cerebral Performance category 1 or 2. MEASUREMENTS AND MAIN RESULTS: In patients treated with therapeutic hypothermia, the median time to reach the target temperature was 5 hrs, with a progressive reduction over the 18 months of data collection. Therapeutic hypothermia had a major positive impact on the outcome of patients with cardiac arrest due to ventricular fibrillation (good outcome in 24 of 43 patients [55.8%] of the therapeutic hypothermia group vs. 11 of 43 patients [25.6%] of the standard resuscitation group, p = .004). The benefit of therapeutic hypothermia was also maintained in patients with shock (good outcome in five of 17 patients of the therapeutic hypothermia group vs. zero of 14 of the standard resuscitation group, p = .027). The outcome after cardiac arrest due to nonventricular fibrillation rhythms was poor and did not differ significantly between the two groups. Therapeutic hypothermia was of particular benefit in patients with short duration of cardiac arrest (<30 mins). CONCLUSIONS: Therapeutic hypothermia for the treatment of postcardiac arrest coma can be successfully implemented in intensive care practice with a major benefit on patient outcome, which appeared to be related to the type and the duration of initial cardiac arrest and seemed maintained in patients with shock. PMID- 16715036 TI - The anti-inflammatory effect of curcumin in an experimental model of sepsis is mediated by up-regulation of peroxisome proliferator-activated receptor-gamma. AB - OBJECTIVE: Although phytochemical curcumin has been shown to possess anti inflammatory properties, it remains unknown whether this agent has any beneficial effects in sepsis. The purpose of this study was to demonstrate whether curcumin protects septic animals and, if so, whether activation of peroxisome proliferator activated receptor (PPAR)-gamma, an anti-inflammatory nuclear receptor, plays any role. DESIGN: Prospective, controlled, and randomized animal study. SETTING: A research institute laboratory. SUBJECTS: Male Sprague-Dawley rats. INTERVENTIONS: A bolus injection of 0.2 micromol of curcumin was given intravenously to male adult rats, followed by continuous infusion of curcumin (0.24 micromol/day) for 3 days via a primed 2-mL mini-pump. The rats were then subjected to sepsis by cecal ligation and puncture (CLP). MEASUREMENTS AND MAIN RESULTS: Serum levels of liver enzymes (alanine aminotransferase and aspartate aminotransferase), lactate, albumin, and tumor necrosis factor (TNF)-alpha were measured at 20 hrs after CLP (i.e., late stage of sepsis). In addition, a 10-day survival curve was conducted following CLP and cecal excision with or without curcumin treatment. Furthermore, macrophages cell line RAW 264.7 cells were treated with curcumin followed by stimulation with endotoxin. TNF-alpha and PPAR-gamma expression were then measured. The results indicate that intravenous administration of curcumin before the onset of sepsis attenuated tissue injury, reduced mortality, and decreased the expression of TNF-alpha in septic animals. Similar results were also found when curcumin was administered after the onset of sepsis. Moreover, the down regulated PPAR-gamma in the liver at 20 hrs after CLP was significantly improved by curcumin treatment. Concurrent administration of curcumin and GW9662, a specific PPAR-gamma antagonist, completely abolished the beneficial effects of curcumin under such conditions. In cultured RAW 264.7 cells, curcumin inhibited endotoxin-induced increases in TNF-alpha expression and markedly up-regulated PPAR-gamma expression without affecting cell viability. Curcumin also prevented morphologic alterations in macrophages induced by endotoxin. CONCLUSIONS: The protective effect of curcumin makes it or its analogues strong candidates as a novel therapy for sepsis. The beneficial effect of curcumin appears to be mediated by up-regulation of nuclear receptor PPAR-gamma. PMID- 16715038 TI - An assessment of the RIFLE criteria for acute renal failure in hospitalized patients. AB - OBJECTIVE: The Acute Dialysis Quality Initiative (ADQI) Group published a consensus definition (the RIFLE criteria) for acute renal failure. We sought to assess the ability of the RIFLE criteria to predict mortality in hospital patients. DESIGN: Retrospective single-center study. SETTING: University affiliated hospital. PATIENTS: All patients admitted to the study hospital between January 2000 and December 2002. Patients were excluded if they were younger than 15 yrs old, were on chronic dialysis, or had kidney transplant or if their length of hospital stay was <24 hrs. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: We included 20,126 patients. Mean age was 64 yrs, 14.7% of patients required intensive care unit admission, and hospital mortality was 8.0%. According to the RIFLE criteria, 9.1% of all patients were in the Risk category for acute renal failure, 5.2% were in the Injury category, and 3.7% were in the Failure category. There was an almost linear increase in hospital mortality from Normal to Failure (Normal, 4.4%; Risk, 15.1%; Injury, 29.2%; and Failure, 41.1%). Multivariate logistic regression analysis showed that all RIFLE criteria were significantly predictive factors for hospital mortality, with an almost linear increase in odds ratios from Risk to Failure (odds ratios, Risk 2.5, Injury 5.4, Failure 10.1). CONCLUSIONS: The RIFLE criteria for acute renal failure classified close to 20% of our study patients as having some degrees of acute impairment in renal function and were useful in predicting their hospital mortality. PMID- 16715037 TI - Role of pneumolysin for the development of acute lung injury in pneumococcal pneumonia. AB - OBJECTIVE: Acute respiratory failure is a significant complication of severe pneumococcal pneumonia. In a mouse model, we observed early-onset lung microvascular leakage after pulmonary infection with Streptococcus pneumoniae, and we hypothesized that the important virulence factor pneumolysin may be the direct causative agent. DESIGN: Controlled, in vivo, ex vivo, and in vitro laboratory study. SETTING: Laboratory. SUBJECTS: Female mice, 8-12 wks old. INTERVENTIONS: Ventilated and blood-free perfused murine lungs were challenged with recombinant pneumolysin via the airways as well as via the vascular bed. In addition, we analyzed the impact of pneumolysin on electrical cell impedance and hydraulic conductivity of human umbilical vein endothelial cell (HUVEC) and alveolar epithelial cell (A549) monolayers. MEASUREMENTS AND MAIN RESULTS: Aerosolized pneumolysin dose-dependently increased capillary permeability with formation of severe lung edema but did not affect pulmonary vascular resistance. Intravascular pneumolysin caused an impressive dose-dependent increase in pulmonary vascular resistance and in lung microvascular permeability. By immunohistochemistry, pneumolysin was detected mainly in endothelial cells of pulmonary arterial vessels, which concomitantly displayed strong vasoconstriction. Moreover, pneumolysin increased permeability of HUVEC and A549 monolayers. Interestingly, immunofluorescence of endothelial cell monolayers exposed to pneumolysin showed gap formation and moderate stress fiber generation. CONCLUSIONS: Pneumolysin may play a central role for early-onset acute lung injury due to severe pneumococcal pneumonia by causing impairment of pulmonary microvascular barrier function and severe pulmonary hypertension. PMID- 16715039 TI - [Prolactin in connective tissue diseases]. AB - This paper presents interactions between prolactin (PRL) and the immune system. We describe the role of PRL in the pathogenesis of rheumatic diseases, particularly connective tissue diseases, including systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), primary Sjogren's syndrome, systemic sclerosis, polymyalgia rheumatica, and seronegative arthritis. We present current opinion on the mechanisms responsible for hyperprolactinemia in SLE patients and the association between hyperprolactinemia and SLE activity and organ involvement. The role of dopamine receptor agonists in the treatment of connective tissue diseases is discussed. PMID- 16715040 TI - [Coronary heart disease in patients with end-stage renal disease on maintenance dialysis. Part I: epidemiology, pathology of atherosclerosis, and diagnosis]. AB - Cardiovascular diseases are the major cause of mortality in patients with end stage renal disease (ESRD) undergoing dialysis. Among ESRD patients the atherosclerotic process progresses more dynamically than in the general population because of exposure to additional, non-traditional risk factors. Diagnosis of coronary artery disease based on history data, physical examination, and additional tests can be difficult due to the coexistence of many disorders and the influence of dialysis treatment. Studies to investigate the pathology of atherosclerosis in patients undergoing dialysis are needed. PMID- 16715042 TI - Map quest. PMID- 16715041 TI - [Coronary heart disease in patients with end-stage renal disease on maintenance dialysis. Part II: prophylaxis and treatment]. AB - Patients with end-stage renal disease (ESRD) and coronary heart disease have a very high risk of cardiovascular mortality. We describe results of recent studies on prevention and treatment strategies. Although the outcomes of revascularization methods, i.e. coronary artery bypass grafting (CABG) and percutaneous coronary intervention (PCI), in ESRD patients are worse than in the general population, CABG and PCI significantly improve survival of the ESRD population. The studies suggest more favorable results with CABG compared with PCI. Patients with a kidney transplant and coronary artery disease (CAD) have better prognosis. In this population, the long-term results of revascularization (CABG and PCI) are comparable. Studies should be focused on optimizing and individualizing revascularization methods, preventing restenosis after PCI, and improving invasive cardiology techniques and anti-aggregation treatments. PMID- 16715043 TI - An accidental breach of a protein's natural defenses. PMID- 16715044 TI - Oskar gains weight. PMID- 16715045 TI - No mercy for messages that mess with the ribosome. PMID- 16715046 TI - Synaptotagmin: fusogenic role for calcium sensor? PMID- 16715048 TI - Determinants of histone H1 mobility and chromatin binding in living cells. AB - The dynamic interaction of chromatin-binding proteins with their nucleosome binding sites is an important element in regulating the structure and function of chromatin in living cells. Here we review the major factors regulating the intranuclear mobility and chromatin binding of the linker histone H1, the most abundant family of nucleosome-binding proteins. The information available reveals that multiple and diverse factors modulate the interaction of H1 with chromatin at both a local and global level. This multifaceted mode of modulating the interaction of H1 with nucleosomes is part of the mechanism that regulates the dynamics of the chromatin fiber in living cells. PMID- 16715049 TI - Cold-adapted archaea. AB - Many archaea are extremophiles. They thrive at high temperatures, at high pressure and in concentrated acidic environments. Nevertheless, the largest proportion and greatest diversity of archaea exist in cold environments. Most of the Earth's biosphere is cold, and archaea represent a significant fraction of the biomass. Although psychrophilic archaea have long been the neglected majority, the study of these microorganisms is beginning to come of age. This review casts a spotlight on the ecology, adaptation biology and unique science that is being realized from studies on cold-adapted archaea. PMID- 16715050 TI - Pushing the envelope: extracytoplasmic stress responses in bacterial pathogens. AB - Despite being nutrient rich, the tissues and fluids of vertebrates are hostile to microorganisms, and most bacteria that attempt to take advantage of this environment are rapidly eliminated by host defences. Pathogens have evolved various means to promote their survival in host tissues, including stress responses that enable bacteria to sense and adapt to adverse conditions. Many different stress responses have been described, some of which are responsive to one or a small number of cues, whereas others are activated by a broad range of insults. The surface layers of pathogenic bacteria directly interface with the host and can bear the brunt of the attack by the host armoury. Several stress systems that respond to perturbations in the microbial cell outside of the cytoplasm have been described and are known collectively as extracytoplasmic or envelope stress responses (ESRs). Here, we review the role of the ESRs in the pathogenesis of Gram-negative bacterial pathogens. PMID- 16715051 TI - Engineered bacteriophage-defence systems in bioprocessing. AB - Bacteriophages (phages) have the potential to interfere with any industry that produces bacteria as an end product or uses them as biocatalysts in the production of fermented products or bioactive molecules. Using microorganisms that drive food bioprocesses as an example, this review will describe a set of genetic tools that are useful in the engineering of customized phage-defence systems. Special focus will be given to the power of comparative genomics as a means of streamlining target selection, providing more widespread phage protection, and increasing the longevity of these industrially important bacteria in the bioprocessing environment. PMID- 16715053 TI - Cerebral preconditioning and ischaemic tolerance. AB - Adaptation is one of physiology's fundamental tenets, operating not only at the level of species, as Darwin proposed, but also at the level of tissues, cells, molecules and, perhaps, genes. During recent years, stroke neurobiologists have advanced a considerable body of evidence supporting the hypothesis that, with experimental coaxing, the mammalian brain can adapt to injurious insults such as cerebral ischaemia to promote cell survival in the face of subsequent injury. Establishing this protective phenotype in response to stress depends on a coordinated response at the genomic, molecular, cellular and tissue levels. Here, I summarize our current understanding of how 'preconditioning' stimuli trigger a cerebroprotective state known as cerebral 'ischaemic tolerance'. PMID- 16715052 TI - Purinergic signalling in neuron-glia interactions. AB - Activity-dependent release of ATP from synapses, axons and glia activates purinergic membrane receptors that modulate intracellular calcium and cyclic AMP. This enables glia to detect neural activity and communicate among other glial cells by releasing ATP through membrane channels and vesicles. Through purinergic signalling, impulse activity regulates glial proliferation, motility, survival, differentiation and myelination, and facilitates interactions between neurons, and vascular and immune system cells. Interactions among purinergic, growth factor and cytokine signalling regulate synaptic strength, development and responses to injury. We review the involvement of ATP and adenosine receptors in neuron-glia signalling, including the release and hydrolysis of ATP, how the receptors signal, the pharmacological tools used to study them, and their functional significance. PMID- 16715054 TI - In vivo imaging of the diseased nervous system. AB - In vivo microscopy is an exciting tool for neurological research because it can reveal how single cells respond to damage of the nervous system. This helps us to understand how diseases unfold and how therapies work. Here, we review the optical imaging techniques used to visualize the different parts of the nervous system, and how they have provided fresh insights into the aetiology and therapeutics of neurological diseases. We focus our discussion on five areas of neuropathology (trauma, degeneration, ischaemia, inflammation and seizures) in which in vivo microscopy has had the greatest impact. We discuss the challenging issues in the field, and argue that the convergence of new optical and non optical methods will be necessary to overcome these challenges. PMID- 16715055 TI - The role of the basal ganglia in habit formation. AB - Many organisms, especially humans, are characterized by their capacity for intentional, goal-directed actions. However, similar behaviours often proceed automatically, as habitual responses to antecedent stimuli. How are goal-directed actions transformed into habitual responses? Recent work combining modern behavioural assays and neurobiological analysis of the basal ganglia has begun to yield insights into the neural basis of habit formation. PMID- 16715056 TI - Seeing at a glance, smelling in a whiff: rapid forms of perceptual decision making. AB - Intuitively, decisions should always improve with more time for the accumulation of evidence, yet psychophysical data show a limit of 200-300 ms for many perceptual tasks. Here, we consider mechanisms that favour such rapid information processing in vision and olfaction. We suggest that the brain limits some types of perceptual processing to short, discrete chunks (for example, eye fixations and sniffs) in order to facilitate the construction of global sensory images. PMID- 16715057 TI - Neuroleptic drugs in dementia: benefits and harm. AB - Neuroleptic (antipsychotic) drugs are often used to treat psychiatric symptoms frequently seen in dementia, but their use is controversial. We present a new meta-analysis to assess the efficacy of these drugs for the treatment of psychiatric symptoms in Alzheimer's disease, and discuss the more limited evidence for their potential benefits in other dementias. We recommend that these treatments be limited to the short-term treatment of psychiatric symptoms associated with serious distress or risk. PMID- 16715059 TI - Regulating inflammation by glucocorticoids. PMID- 16715058 TI - Mainstreaming the alternative. PMID- 16715060 TI - Origins of the T(H)1-T(H)2 model: a personal perspective. AB - Robert L. Coffman recounts how his work on immunoglobulin E regulation along with data from Tim Mosmann on the functional heterogeneity of T cell clones led to the T helper type 1-T helper type 2 hypothesis. PMID- 16715061 TI - Medicine on a need-to-know basis. AB - Disease-oriented, introductory medical curricula can help overcome educational and institutional barriers that separate aspiring translational scientists in PhD programs from the world of medicine. PMID- 16715062 TI - Cytosolic detection of flagellin: a deadly twist. PMID- 16715063 TI - Plasmacytoid DCs and T(reg) cells: casual acquaintance or monogamous relationship? PMID- 16715064 TI - Stress management for T helper differentiation. PMID- 16715065 TI - Pin-ning down immune responses to RNA viruses. PMID- 16715066 TI - TGF-beta, a 'double agent' in the immune pathology war. PMID- 16715068 TI - Is RNA interference involved in intrinsic antiviral immunity in mammals? AB - RNA interference constitutes a key component of the innate immune response to viral infection in both plants and invertebrate animals and has been postulated to have a similar protective function in mammals. This perspective reviews the available data addressing whether RNA interference forms part of the mammalian innate immune response and concludes that the popular hypothesis in favor of that possibility remains far from proven and may not be valid. PMID- 16715071 TI - Molecular detection of Campylobacter jejuni in archival cases of acute appendicitis. AB - The role of enteric bacteria in the pathogenesis of acute appendicitis is a controversial subject. Campylobacter jejuni has been previously demonstrated in a minority of cases of acute appendicitis using microbiological or immunohistochemical methods, notably in cases where inflammation was limited to the mucosa/submucosa. Our goal was to evaluate cases of acute appendicitis for C. jejuni DNA using molecular methods, and to compare our findings to the histologic features. In total, 50 archival cases of acute appendicitis were selected, and PCR was performed using primers targeting a 286-bp fragment of the mapA gene specific to C. jejuni. Twenty histologically unremarkable appendectomy specimens served as negative controls. Cases were reviewed with attention to particular histological features including mucosal ulceration, cryptitis, depth of inflammatory infiltrate, and the presence of mural necrosis. Of acute appendicitis cases, 22% (11/50) were positive for C. jejuni DNA by PCR analysis. Control cases were negative for C. jejuni DNA. All patients presented with signs and symptoms typical of acute appendicitis. Of the C. jejuni positive cases, only 27% contained acute inflammation limited to the mucosa/submucosa, whereas the remainder had mural or transmural inflammation; therefore, the histological features of C. jejuni-positive acute appendicitis cases were indistinguishable from C. jejuni-negative cases. In summary, C. jejuni DNA was detected in a significant percentage (22%) of acute appendicitis cases, a much higher percentage than previous studies using other methodologies. As C. jejuni is an enteric pathogen that does not exist as a commensal or nonpathogenic organism, the presence of C. jejuni DNA implies current or recent infection. Further study is needed to determine whether the presence of C. jejuni DNA in acute appendicitis indicates appendiceal involvement by C. jejuni enteritis, or if there is a true causative role for C. jejuni in acute appendicitis. PMID- 16715072 TI - Granulomatous pyelitis associated with urinary obstruction: a comprehensive clinicopathologic study. AB - Urinary obstruction is rarely associated with a distinct granulomatous inflammation, which involves the pyelocalyceal system and closely simulates infectious conditions including tuberculosis. Its clinicopathologic features, however, have not been adequately studied since there are only seven isolated reported cases. In a comprehensive study of 112 kidney specimens with urinary obstruction, we identified five cases of granulomatous pyelitis. The features of these cases were detailed and compared with the previously reported cases. Among the five identified subjects, three patients had history of urolithiasis and two had ureteral stenosis and all had stent placement 7 weeks to 12 years before nephrectomy for relief of the unilateral urinary obstruction. The age distribution was between 38 and 81 years. Two had end-stage renal disease or chronic renal failure. The pyelocaliceal system showed frank hydronephrosis (1 case) or partial dilatation (4 cases) and contained cheesy and gritty material in its lumen. Each case showed severe granulomatous inflammation, which was limited to the pelvic wall and closely associated with calcified debris, necrotic inflammatory cells, and material consistent with Tamm-Horsfall protein. The kidney showed chronic tubulointerstitial nephritis but without granulomas. Cultures of urine, blood, and the renal pelvic content, and special stains of tissue sections did not show fungi or mycobacteria in any case. Many of these features were also observed in previously reported cases. Granulomatous pyelitis is a rare but distinct cliniocopathologic entity characterized by severe noninfectious granulomatous inflammation limited to the renal pelvis, which is uniformely asociated with urinary obstruction and pyelocalyceal dilatation and may develop in response to accumulated calcified material in the renal pelvis. Awareness of this entity and its characteristic clinicopathologic features also helps eliminate an infectious etiology with obvious treatment and prognostic implications. PMID- 16715073 TI - High throughput tissue microarray analysis of FHIT expression in diffuse large cell B-cell lymphoma from Saudi Arabia. AB - Recent studies have suggested a potential prognostic role of alterations of the fragile histidine triad (FHIT) gene in diffuse large B-cell lymphoma. To evaluate possible mechanisms of FHIT inactivation and to further clarify its potential prognostic relevance, we analyzed a set of 114 diffuse large B-cell lymphoma with clinical follow-up information. Tissue microarrays were analyzed by immunohistochemistry for protein expression, and corresponding DNA samples were analyzed for FHIT promotor hypermethlyation. Reduced or absent FHIT expression was found in 75 of 114 diffuse large B-cell lymphoma (66%), but was unrelated to clinical tumor stage or patient prognosis. FHIT promotor hypermethylation was observed in 29 of 93 (23%) interpretable diffuse large B-cell lymphoma. Hypermethylation was not significantly correlated to protein expression loss, which could be explained by competing mechanisms for FHIT inactivation in a substantial fraction of non FHIT hypermethylated diffuse large B-cell lymphoma. Hypermethylation was significantly associated with poor prognosis of diffuse large B-cell lymphoma patients and predominantly seen in nongerminal center diffuse large B-cell lymphoma (27%), but less frequent (13%) in germinal center diffuse large B-cell lymphoma. In summary, these data suggest that promotor hypermethylation is responsible for reduced FHIT expression in a substantial subset of diffuse large B-cell lymphoma, which is primarily composed of nongerminal center subtype with poor patient prognosis. PMID- 16715074 TI - Dual selection enhances the signaling specificity of a variant of the quorum sensing transcriptional activator LuxR. AB - The transcription factor LuxR activates gene expression in response to binding the signaling molecule 3-oxo-hexanoyl-homoserine lactone (3OC6HSL), an acyl-HSL with a carbonyl substituent at the third carbon of the acyl chain. We previously described a LuxR variant, LuxR-G2E, that activates gene expression by binding a broader range of acyl-HSLs, including straight-chain acyl-HSLs to which LuxR does not respond. Here, we use a dual positive-negative selection system to identify a variant of LuxR-G2E that retains the response to straight-chain acyl-HSLs, but no longer responds to 3OC6HSL. A single mutation, R67M, reduces LuxR-G2E's response to acyl-HSLs having a carbonyl substituent at the third carbon of the acyl chain. This specificity-enhancing mutation would not have been identified by positive selection alone. The dual selection system provides a rapid and reliable method for identifying LuxR variants that have or lack the desired response to a given set of acyl-HSL signals. LuxR variants with altered signaling specificities might become useful components for constructing artificial cell-cell communication systems that program population level behaviors. PMID- 16715075 TI - Processive bidirectional motion of dynein-dynactin complexes in vitro. AB - Cytoplasmic dynein is the primary molecular motor responsible for transport of vesicles, organelles, proteins and RNA cargoes from the periphery of the cell towards the nucleus along the microtubule cytoskeleton of eukaryotic cells. Dynactin, a large multi-subunit activator of dynein, docks cargo to the motor and may enhance dynein processivity. Here, we show that individual fluorescently labelled dynein-dynactin complexes exhibit bidirectional and processive motility towards both the plus and minus ends of microtubules. The dependence of this activity on substrate ATP concentration, nucleotide analogues and inhibitors suggests that bidirectional motility is an active energy-transduction property of dynein-dynactin motor mechano-chemistry. The unique motility characteristics observed may reflect the flexibility of the dynein structure that leads to an enhanced ability to navigate around obstacles in the cell. PMID- 16715076 TI - p63 regulates an adhesion programme and cell survival in epithelial cells. AB - p63 is critical for epithelial development yet little is known about the transcriptional programmes it regulates. By characterising transcriptional changes and cellular effects following modulation of p63 expression, we have defined a vital role for p63 in cellular adhesion. Knockdown of p63 expression caused downregulation of cell adhesion-associated genes, cell detachment and anoikis in mammary epithelial cells and keratinocytes. Conversely, overexpression of the TAp63gamma or deltaNp63alpha isoforms of p63 upregulated cell adhesion molecules, increased cellular adhesion and conferred resistance to anoikis. Apoptosis induced by loss of p63 was rescued by signalling downstream of beta4 integrin. Our results implicate p63 as a key regulator of cellular adhesion and survival in basal cells of the mammary gland and other stratified epithelial tissues. PMID- 16715077 TI - Plexin-A1 and its interaction with DAP12 in immune responses and bone homeostasis. AB - Semaphorins and their receptors have diverse functions in axon guidance, organogenesis, vascularization and/or angiogenesis, oncogenesis and regulation of immune responses. The primary receptors for semaphorins are members of the plexin family. In particular, plexin-A1, together with ligand-binding neuropilins, transduces repulsive axon guidance signals for soluble class III semaphorins, whereas plexin-A1 has multiple functions in chick cardiogenesis as a receptor for the transmembrane semaphorin, Sema6D, independent of neuropilins. Additionally, plexin-A1 has been implicated in dendritic cell function in the immune system. However, the role of plexin-A1 in vivo, and the mechanisms underlying its pleiotropic functions, remain unclear. Here, we generated plexin-A1-deficient (plexin-A1(-/-)) mice and identified its important roles, not only in immune responses, but also in bone homeostasis. Furthermore, we show that plexin-A1 associates with the triggering receptor expressed on myeloid cells-2 (Trem-2), linking semaphorin-signalling to the immuno-receptor tyrosine-based activation motif (ITAM)-bearing adaptor protein, DAP12. These findings reveal an unexpected role for plexin-A1 and present a novel signalling mechanism for exerting the pleiotropic functions of semaphorins. PMID- 16715078 TI - Molecular architecture of a kinetochore-microtubule attachment site. AB - Kinetochore attachment to spindle microtubule plus-ends is necessary for accurate chromosome segregation during cell division in all eukaryotes. The centromeric DNA of each chromosome is linked to microtubule plus-ends by eight structural protein complexes. Knowing the copy number of each of these complexes at one kinetochore-microtubule attachment site is necessary to understand the molecular architecture of the complex, and to elucidate the mechanisms underlying kinetochore function. We have counted, with molecular accuracy, the number of structural protein complexes in a single kinetochore-microtubule attachment using quantitative fluorescence microscopy of GFP-tagged kinetochore proteins in the budding yeast Saccharomyces cerevisiae. We find that relative to the two Cse4p molecules in the centromeric histone, the copy number ranges from one or two for inner kinetochore proteins such as Mif2p, to 16 for the DAM-DASH complex at the kinetochore-microtubule interface. These counts allow us to visualize the overall arrangement of a kinetochore-microtubule attachment. As most of the budding yeast kinetochore proteins have homologues in higher eukaryotes, including humans, this molecular arrangement is likely to be replicated in more complex kinetochores that have multiple microtubule attachments. PMID- 16715079 TI - Impaired light masking in dopamine D2 receptor-null mice. AB - Disruption of overt circadian rhythms can occur without influencing the endogenous pacemaker, the so-called 'masking' effect classically elicited by light. As the physiological pathways involved in light masking remain elusive, we analyzed mice lacking the dopamine D2 receptor. Although circadian rhythmicity was normal, D2R-null mice showed a markedly deficient light masking response, indicating that D2R-mediated signaling is an essential component of the neuronal pathways leading to light masking of circadian rhythms. PMID- 16715080 TI - Adaptation of gender derived from biological motion. AB - Human observers adapted to complex biological motions that distinguish males from females: viewing the gait of one gender biased judgments of subsequent gaits toward the opposite gender. This adaptation was not simply due to local features of the stimuli but instead relied upon the global motion of the figures. These results suggest the existence of neurons selective for gender and demonstrate that gender-from-motion judgments are not fixed but depend upon recent viewing history. PMID- 16715083 TI - Robust free-standing nanomembranes of organic/inorganic interpenetrating networks. AB - Hybrid sol-gel materials have been a subject of intensive research during the past decades because these nanocomposites combine the versatility of organic polymers with the superior physical properties of glass. Here, we report the synthesis, by spin coating, of hybrid interpenetrating networks in the form of free-standing nanomembrane (around 35-nm thick) with unprecedented macroscopic size and characteristics. The quasi-2D interpenetration of the organic and inorganic networks brings to these materials a unique combination of properties that are not usually compatible within the same film: macroscopic robustness and homogeneity, nanoscale thickness, mechanical strength, high flexibility and optical transparency. Interestingly, such free-standing nanofilms of macroscopic size can seal large openings, are strong enough to hold amounts of liquid 70,000 times heavier than their own weight, and are flexible enough to reversibly pass through holes 30,000 times smaller than their own size. PMID- 16715082 TI - Midline radial glia translocation and corpus callosum formation require FGF signaling. AB - Midline astroglia in the cerebral cortex develop earlier than other astrocytes through mechanisms that are still unknown. We show that radial glia in dorsomedial cortex retract their apical endfeet at midneurogenesis and translocate to the overlaying pia, forming the indusium griseum. These cells require the fibroblast growth factor receptor 1 (Fgfr1) gene for their precocious somal translocation to the dorsal midline, as demonstrated by inactivating the Fgfr1 gene in radial glial cells and by RNAi knockdown of Fgfr1 in vivo. Dysfunctional astroglial migration underlies the callosal dysgenesis in conditional Fgfr1 knockout mice, suggesting that precise targeting of astroglia to the cortex has unexpected roles in axon guidance. FGF signaling is sufficient to induce somal translocation of radial glial cells throughout the cortex; furthermore, the targeting of astroglia to dorsolateral cortex requires FGFr2 signaling after neurogenesis. Hence, FGFs have an important role in the transition from radial glia to astrocytes by stimulating somal translocation of radial glial cells. PMID- 16715084 TI - Xenon as a probe for minority sites on solid surfaces. AB - Atomic-level microscopies have proved useful to map solid-surface sites directly, but, because of their lack of chemical specificity, they are less adept at identifying unique chemical activity on those sites. Here we present a dual titration approach developed to probe minority sites on solid surfaces with unique chemical properties of potential relevance to heterogeneous catalysis. Our methodology involves the initial dosing of a chemical probe such as carbon monoxide or ammonia to drive its selective adsorption onto specific sites with particular chemical activity, and the subsequent adsorption of xenon to help identify the nature of those sites. A combination of photoelectron and temperature-programmed desorption spectroscopies are used to characterize the Xe adsorption. The chemistry of oxygen-modified Ni(110) single crystals was probed to test this technique. It was observed that whereas CO does not discriminate among the various sites present on those surfaces, ammonia binds preferentially to the end of -Ni-O rows and modifies their local electrostatic potential. In addition, it was determined that adsorbed CO aids in a reversible surface reconstruction involving the coalescence of fragmented surface -Ni-O rows at high (>350 K) temperatures. PMID- 16715081 TI - Ascl1 and Gsh1/2 control inhibitory and excitatory cell fate in spinal sensory interneurons. AB - Sensory information from the periphery is integrated and transduced by excitatory and inhibitory interneurons in the dorsal spinal cord. Recent studies have identified a number of postmitotic factors that control the generation of these sensory interneurons. We show that Gsh1/2 and Ascl1 (Mash1), which are expressed in sensory interneuron progenitors, control the choice between excitatory and inhibitory cell fates in the developing mouse spinal cord. During the early phase of neurogenesis, Gsh1/2 and Ascl1 coordinately regulate the expression of Tlx3, which is a critical postmitotic determinant for dorsal glutamatergic sensory interneurons. However, at later developmental times, Ascl1 controls the expression of Ptf1a in dIL(A) progenitors to promote inhibitory neuron differentiation while at the same time upregulating Notch signaling to ensure the proper generation of dIL(B) excitatory neurons. We propose that this switch in Ascl1 function enables the cogeneration of inhibitory and excitatory sensory interneurons from a common pool of dorsal progenitors. PMID- 16715085 TI - Reversible active switching of the mechanical properties of a peptide film at a fluid-fluid interface. AB - Designer peptides have recently been developed as building blocks for novel self assembled materials with stimuli-responsive properties. To date, such materials have been based on self-assembly in bulk aqueous solution or at solid-fluid interfaces. We have designed a 21-residue peptide, AM1, as a stimuli-responsive surfactant that switches molecular architectures at a fluid-fluid interface in response to changes in bulk aqueous solution composition. In the presence of divalent zinc at neutral pH, the peptide forms a mechanically strong 'film state'. In the absence of metal ions or at acid pH, the peptide adsorbs to form a mobile 'detergent state'. The two interfacial states can be actively and reversibly switched. Switching between the two states by a change in pH or the addition of a chelating agent leads to rapid emulsion coalescence or foam collapse. This work introduces a new class of surfactants that offer an environmentally friendly approach to control the stability of interfaces in foams, emulsions and fluid-fluid interfaces more generally. PMID- 16715086 TI - Complex precipitation pathways in multicomponent alloys. AB - One usual way to strengthen a metal is to add alloying elements and to control the size and the density of the precipitates obtained. However, precipitation in multicomponent alloys can take complex pathways depending on the relative diffusivity of solute atoms and on the relative driving forces involved. In Al-Zr Sc alloys, atomic simulations based on first-principle calculations combined with various complementary experimental approaches working at different scales reveal a strongly inhomogeneous structure of the precipitates: owing to the much faster diffusivity of Sc compared with Zr in the solid solution, and to the absence of Zr and Sc diffusion inside the precipitates, the precipitate core is mostly Sc rich, whereas the external shell is Zr-rich. This explains previous observations of an enhanced nucleation rate in Al-Zr-Sc alloys compared with binary Al-Sc alloys, along with much higher resistance to Ostwald ripening, two features of the utmost importance in the field of light high-strength materials. PMID- 16715087 TI - CB1 cannabinoid receptor antagonism: a new strategy for the treatment of liver fibrosis. AB - Hepatic fibrosis, the common response associated with chronic liver diseases, ultimately leads to cirrhosis, a major public health problem worldwide. We recently showed that activation of hepatic cannabinoid CB2 receptors limits progression of experimental liver fibrosis. We also found that during the course of chronic hepatitis C, daily cannabis use is an independent predictor of fibrosis progression. Overall, these results suggest that endocannabinoids may drive both CB2-mediated antifibrogenic effects and CB2-independent profibrogenic effects. Here we investigated whether activation of cannabinoid CB1 receptors (encoded by Cnr1) promotes progression of fibrosis. CB1 receptors were highly induced in human cirrhotic samples and in liver fibrogenic cells. Treatment with the CB1 receptor antagonist SR141716A decreased the wound-healing response to acute liver injury and inhibited progression of fibrosis in three models of chronic liver injury. We saw similar changes in Cnr1-/- mice as compared to wild type mice. Genetic or pharmacological inactivation of CB1 receptors decreased fibrogenesis by lowering hepatic transforming growth factor (TGF)-beta1 and reducing accumulation of fibrogenic cells in the liver after apoptosis and growth inhibition of hepatic myofibroblasts. In conclusion, our study shows that CB1 receptor antagonists hold promise for the treatment of liver fibrosis. PMID- 16715088 TI - Generation of C5a in the absence of C3: a new complement activation pathway. AB - Complement-mediated tissue injury in humans occurs upon deposition of immune complexes, such as in autoimmune diseases and acute respiratory distress syndrome. Acute lung inflammatory injury in wild-type and C3-/- mice after deposition of IgG immune complexes was of equivalent intensity and was C5a dependent, but injury was greatly attenuated in Hc-/- mice (Hc encodes C5). Injury in lungs of C3-/- mice and C5a levels in bronchoalveolar lavage (BAL) fluids from these mice were greatly reduced in the presence of antithrombin III (ATIII) or hirudin but were not reduced in similarly treated C3+/+ mice. Plasma from C3-/- mice contained threefold higher levels of thrombin activity compared to plasma from C3+/+ mice. There were higher levels of F2 mRNA (encoding prothrombin) as well as prothrombin and thrombin protein in liver of C3-/- mice compared to C3+/+ mice. A potent solid-phase C5 convertase was generated using plasma from either C3+/+ or C3-/- mice. Human C5 incubated with thrombin generated C5a that was biologically active. These data suggest that, in the genetic absence of C3, thrombin substitutes for the C3-dependent C5 convertase. This linkage between the complement and coagulation pathways may represent a new pathway of complement activation. PMID- 16715089 TI - Osteoclasts degrade endosteal components and promote mobilization of hematopoietic progenitor cells. AB - Here we investigated the potential role of bone-resorbing osteoclasts in homeostasis and stress-induced mobilization of hematopoietic progenitors. Different stress situations induced activity of osteoclasts (OCLs) along the stem cell-rich endosteum region of bone, secretion of proteolytic enzymes and mobilization of progenitors. Specific stimulation of OCLs with RANKL recruited mainly immature progenitors to the circulation in a CXCR4- and MMP-9-dependent manner; however, RANKL did not induce mobilization in young female PTPepsilon knockout mice with defective OCL bone adhesion and resorption. Inhibition of OCLs with calcitonin reduced progenitor egress in homeostasis, G-CSF mobilization and stress situations. RANKL-stimulated bone-resorbing OCLs also reduced the stem cell niche components SDF-1, stem cell factor (SCF) and osteopontin along the endosteum, which was associated with progenitor mobilization. Finally, the major bone-resorbing proteinase, cathepsin K, also cleaved SDF-1 and SCF. Our findings indicate involvement of OCLs in selective progenitor recruitment as part of homeostasis and host defense, linking bone remodeling with regulation of hematopoiesis. PMID- 16715090 TI - Intravenous immunoglobulin ameliorates ITP via activating Fc gamma receptors on dendritic cells. AB - Despite a more than 20-year experience of therapeutic benefit, the relevant molecular and cellular targets of intravenous immunoglobulin (IVIg) in autoimmune disease remain unclear. Contrary to the prevailing theories of IVIg action in autoimmunity, we show that IVIg drives signaling through activating Fc gamma receptors (Fc gammaR) in the amelioration of mouse immune thrombocytopenic purpura (ITP). The actual administration of IVIg was unnecessary because as few as 10(5) IVIg-treated cells could, upon adoptive transfer, ameliorate ITP. IVIg did not interact with the inhibitory Fc gammaRIIB on the initiator cell, although Fc gammaRIIB does have a role in the late phase of IVIg action. Notably, only IVIg-treated CD11c+ dendritic cells could mediate these effects. We hypothesize that IVIg forms soluble immune complexes in vivo that prime dendritic-cell regulatory activity. In conclusion, the clinical effects of IVIg in ameliorating ITP seem to involve the acute interaction of IVIg with activating Fc gammaR on dendritic cells. PMID- 16715091 TI - Oral CD3-specific antibody suppresses autoimmune encephalomyelitis by inducing CD4+ CD25- LAP+ T cells. AB - A major goal of immunotherapy for autoimmune diseases and transplantation is induction of regulatory T cells that mediate immunologic tolerance. The mucosal immune system is unique, as tolerance is preferentially induced after exposure to antigen, and induction of regulatory T cells is a primary mechanism of oral tolerance. Parenteral administration of CD3-specific monoclonal antibody is an approved therapy for transplantation in humans and is effective in autoimmune diabetes. We found that orally administered CD3-specific antibody is biologically active in the gut and suppresses autoimmune encephalomyelitis both before induction of disease and at the height of disease. Orally administered CD3 specific antibody induces CD4+ CD25- LAP+ regulatory T cells that contain latency associated peptide (LAP) on their surface and that function in vitro and in vivo through a TGF-beta-dependent mechanism. These findings identify a new immunologic approach that is widely applicable for the treatment of human autoimmune conditions. PMID- 16715092 TI - Synaptic scaffolding protein Homer1a protects against chronic inflammatory pain. AB - Glutamatergic signaling and intracellular calcium mobilization in the spinal cord are crucial for the development of nociceptive plasticity, which is associated with chronic pathological pain. Long-form Homer proteins anchor glutamatergic receptors to sources of calcium influx and release at synapses, which is antagonized by the short, activity-dependent splice variant Homer1a. We show here that Homer1a operates in a negative feedback loop to regulate the excitability of the pain pathway in an activity-dependent manner. Homer1a is rapidly and selectively upregulated in spinal cord neurons after peripheral inflammation in an NMDA receptor-dependent manner. Homer1a strongly attenuates calcium mobilization as well as MAP kinase activation induced by glutamate receptors and reduces synaptic contacts on spinal cord neurons that process pain inputs. Preventing activity-induced upregulation of Homer1a using shRNAs in mice in vivo exacerbates inflammatory pain. Thus, activity-dependent uncoupling of glutamate receptors from intracellular signaling mediators is a novel, endogenous physiological mechanism for counteracting sensitization at the first, crucial synapse in the pain pathway. Furthermore, we observed that targeted gene transfer of Homer1a to specific spinal segments in vivo reduces inflammatory hyperalgesia. Thus, Homer1 function is crucially involved in pain plasticity and constitutes a promising therapeutic target for the treatment of chronic inflammatory pain. PMID- 16715093 TI - PUF proteins bind Pop2p to regulate messenger RNAs. AB - PUF proteins, a family of RNA-binding proteins, interact with the 3' untranslated regions (UTRs) of specific mRNAs to control their translation and stability. PUF protein action is commonly correlated with removal of the poly(A) tail of target mRNAs. Here, we focus on how PUF proteins enhance deadenylation and mRNA decay. We show that a yeast PUF protein physically binds Pop2p, which is a component of the Ccr4p-Pop2p-Not deadenylase complex, and that Pop2p is required for PUF repression activity. By binding Pop2p, the PUF protein simultaneously recruits the Ccr4p deadenylase and two other enzymes involved in mRNA regulation, Dcp1p and Dhh1p. We reconstitute regulated deadenylation in vitro and demonstrate that the PUF-Pop2p interaction is conserved in yeast, worms and humans. We suggest that the PUF-Pop2p interaction underlies regulated deadenylation, mRNA decay and repression by PUF proteins. PMID- 16715094 TI - X-ray structure of influenza virus NS1 effector domain. AB - The nonstructural protein NS1 of influenza virus is an antagonist of host immune responses and is implicated in virulence. It has two domains, an N-terminal double-stranded RNA-binding domain (RBD) and an effector domain crucial for RBD function, for nuclear export and for sequestering messenger RNA-processing proteins. Here we present the crystallographic structure of the effector domain, which has a novel fold and suggests mechanisms for increased virulence in H5N1 strains. PMID- 16715095 TI - Importin-alpha-16 is a translocon-associated protein involved in sorting membrane proteins to the nuclear envelope. AB - A viral inner nuclear membrane-sorting motif sequence (INM-SM) was used to identify proteins that recognize integral membrane proteins destined for the INM. Herein we describe importin-alpha-16, a membrane-associated isoform of Spodoptera frugiperda importin-alpha that contains the C-terminal amino acid residues comprising armadillo helical-repeat domains 7-10. In the endoplasmic reticulum (ER) membrane, importin-alpha-16 is adjacent to the translocon protein Sec61alpha. Importin-alpha-16 cross-links to the INM-SM sequence as it emerges from the ribosomal tunnel and remains adjacent to the INM-SM after INM-SM integration into the ER membrane and release from the translocon. Cross-linking results suggest that importin-alpha-16 discriminates between INM- and non-INM directed proteins. Thus, it seems that during and after cotranslational membrane integration, importin-alpha-16 is involved in the trafficking of integral membrane proteins to the INM. PMID- 16715096 TI - Evaluating and improving power in whole-genome association studies using fixed marker sets. AB - Emerging technologies make it possible for the first time to genotype hundreds of thousands of SNPs simultaneously, enabling whole-genome association studies. Using empirical genotype data from the International HapMap Project, we evaluate the extent to which the sets of SNPs contained on three whole-genome genotyping arrays capture common SNPs across the genome, and we find that the majority of common SNPs are well captured by these products either directly or through linkage disequilibrium. We explore analytical strategies that use HapMap data to improve power of association studies conducted with these fixed sets of markers and show that limited inclusion of specific haplotype tests in association analysis can increase the fraction of common variants captured by 25-100%. Finally, we introduce a Bayesian approach to association analysis by weighting the likelihood of each statistical test to reflect the number of putative causal alleles to which it is correlated. PMID- 16715097 TI - Noise in protein expression scales with natural protein abundance. AB - Noise in gene expression is generated at multiple levels, such as transcription and translation, chromatin remodeling and pathway-specific regulation. Studies of individual promoters have suggested different dominating noise sources, raising the question of whether a general trend exists across a large number of genes and conditions. We examined the variation in the expression levels of 43 Saccharomyces cerevisiae proteins, in cells grown under 11 experimental conditions. For all classes of genes and under all conditions, the expression variance was approximately proportional to the mean; the same scaling was observed at steady state and during the transient responses to the perturbations. Theoretical analysis suggests that this scaling behavior reflects variability in mRNA copy number, resulting from random 'birth and death' of mRNA molecules or from promoter fluctuations. Deviation of coexpressed genes from this general trend, including high noise in stress-related genes and low noise in proteasomal genes, may indicate fluctuations in pathway-specific regulators or a differential activation pattern of the underlying gene promoters. PMID- 16715098 TI - Mutations in GLIS3 are responsible for a rare syndrome with neonatal diabetes mellitus and congenital hypothyroidism. AB - We recently described a new neonatal diabetes syndrome associated with congenital hypothyroidism, congenital glaucoma, hepatic fibrosis and polycystic kidneys. Here, we show that this syndrome results from mutations in GLIS3, encoding GLI similar 3, a recently identified transcription factor. In the original family, we identified a frameshift mutation predicted to result in a truncated protein. In two other families with an incomplete syndrome, we found that affected individuals harbor deletions affecting the 11 or 12 5'-most exons of the gene. The absence of a major transcript in the pancreas and thyroid (deletions from both families) and an eye-specific transcript (deletion from one family), together with residual expression of some GLIS3 transcripts, seems to explain the incomplete clinical manifestations in these individuals. GLIS3 is expressed in the pancreas from early developmental stages, with greater expression in beta cells than in other pancreatic tissues. These results demonstrate a major role for GLIS3 in the development of pancreatic beta cells and the thyroid, eye, liver and kidney. PMID- 16715099 TI - Evaluating coverage of genome-wide association studies. AB - Genome-wide association studies involving hundreds of thousands of SNPs in thousands of cases and controls are now underway. The first of many analytical challenges in these studies involves the choice of SNPs to genotype. It is not practical to construct a different panel of tag SNPs for each study, so the first generation of genome-wide scans will use predefined, commercially available marker panels, which will in part dictate their success or failure. We compare different approaches in use today, and show that although many of them provide substantial coverage of common variation in non-African populations, the precise extent is strongly dependent on the frequencies of alleles of interest and on specific considerations of study design. Overall, despite substantial differences in genotyping technologies, marker selection strategies and number of markers assayed, the first-generation high-throughput platforms all offer similar levels of genome coverage. PMID- 16715100 TI - Neutrophil direction sensing and superoxide production linked by the GTPase activating protein GIT2. AB - In neutrophils, superoxide anion production generally accompanies chemotaxis and functions in killing invading pathogens. The GIT2 GTPase-activating protein binds to the guanine nucleotide-exchange factor alphaPIX. Here we show that GIT2 was necessary for directional chemotaxis and for the suppression of superoxide production in G protein-coupled receptor-stimulated neutrophils. GIT2 was also necessary for the orientation of superoxide production toward chemoattractant sources. GIT2 suppressed the activity of ADP ribosylation factor 1 and was a component of the Gbetagamma subunit-mediated direction-sensing machinery 'downstream' of G protein-coupled receptor signaling. This study establishes a function for GIT2 in linking chemotaxis and superoxide production in neutrophils and shows that loss of GIT2 in vivo leads to an immunodeficient state. PMID- 16715102 TI - Cytokine-induced neutrophil chemoattractant-1 is released by the noninjured liver in a rat acute-phase model. AB - The source of serum cytokine-induced neutrophil chemoattractant (CINC-1) and consequences of its presence in the tissue of synthesis have not been clearly elucidated under acute-phase situation. To pursue this question, turpentine oil (TO) was intramuscularly injected into rats, and RNA and local protein levels of acute-phase cytokines and of CINC-1 were studied in the TO injected gluteal muscle, as well as in noninjured muscle, in the liver, kidney, lung and spleen. The serum levels of acute-phase mediators and of CINC-1 were measured together with total leukocyte subpopulations. Recruitment of inflammatory cells in muscle and in the other organs was investigated by quantitative immunohistochemical methods. The effect of acute-phase mediators, including interferon gamma (IFN gamma) on the synthesis of CINC-1 in cultured hepatocytes was also investigated at the RNA and protein level. We found that the sera of the TO-treated rats contained elevated levels of IL-6, IL-1beta and CINC-1. Increased serum levels of IFN-gamma were also observed not only in the injured muscle but also and to a higher extent in the liver. However, while neutrophils and mononuclear phagocytes were found in the injured muscle, no inflammatory cells were detected at the non 'inflamed' site, namely, the liver or in the other organs. In vitro, treatment of cultured hepatocytes with IL-1beta led to elevated CINC-1 gene expression. This was true to a lesser extent upon IL-6 and tumor necrosis factor (TNF-alpha) exposure. Interestingly, IFN-gamma did not effect CINC-1 gene expression. These results indicate that CINC-1 behaves as an acute-phase protein and its expression is inducible in hepatocytes. However, CINC-1-production in the liver does not lead to recruitment of inflammatory cells into the organ. PMID- 16715101 TI - Interleukin 15-dependent crosstalk between conventional and plasmacytoid dendritic cells is essential for CpG-induced immune activation. AB - The function of interleukin 15 (IL-15) in unmethylated CpG oligodeoxynucleotide (CpG)-induced immune responses remains unknown. Here, in response to CpG, both wild-type and natural killer cell-depleted mice produced IL-12 and became resistant to a lethal dose of Listeria monocytogenes. In contrast, CpG-treated IL 15-deficient mice produced little IL-12 and succumbed to L. monocytogenes. CpG stimulated conventional dendritic cells (cDCs) were the main producers of both IL 15 and IL-12, but cDCs did not produce IL-12 in the absence of plasmacytoid DCs (pDCs). The cDC-derived IL-15 induced CD40 expression by cDCs. Interaction between CD40 on cDCs and CD40 ligand on pDCs led to IL-12 production by cDCs. Thus, IL-15-dependent crosstalk between cDCs and pDCs is essential for CpG induced immune activation. PMID- 16715103 TI - Assessment of converting from intravenous to oral administration of cyclosporin A in pediatric allogeneic hematopoietic stem cell transplant recipients. AB - We studied the administration method during a transition period from continuous intravenous (i.v.) infusion to oral administration of cyclosporin A (CsA). Thirty two pediatric hematopoietic stem cell transplant (HSCT) recipients, between the ages of 8 months and 15.6 years (median 7.1 years) participated in this study. The pharmacokinetic properties of CsA was evaluated during the transition period from i.v. to oral CsA. The daily oral dose of CsA was three times higher than the i.v. dose. Oral dosing began immediately after the continuous infusion was discontinued. Whole-blood CsA concentrations were measured by a monoclonal fluorescence polarization immunoassay (FPIA). The mean+/-s.d. value of bioavailability (F), maximum concentration (C(max)), half-life (t(1/2)) of CsA were 43.1+/-14.4%, 1135.3+/-340.6 ng/ml and 3.1+/-1.2 h, respectively. Mean clearance (CL)+/-s.d. was 480.9+/-103.7, 414.9+/-137.1 and 320+/-51.8 ml/h/kg in patients <20, 20-40 and >40 kg of body weight, respectively. The CsA CL of younger children was significantly greater than for older children (P=0.044). CsA trough levels were maintained within the therapeutic range throughout the transition period. Therefore, our findings suggest that the immediate administration of an oral formulation, after discontinuation of the continuous infusion, would be practical and effective for routine clinical use. PMID- 16715105 TI - A case of hepatic graft-versus-host disease complicating autologous stem cell transplantation for relapsed Hodgkin's disease. PMID- 16715104 TI - Haematopoietic cell transplantation (HCT) in combination with enzyme replacement therapy (ERT) in patients with Hurler syndrome. AB - Hurler syndrome (MPS 1H) is the severe form of mucopolysaccharidosis type 1 (MPS 1). Haematopoietic cell transplantation (HCT) is the treatment of choice, but carries a high incidence of graft failure and morbidity. The use of enzyme replacement therapy (ERT) might improve the clinical signs and symptoms before HCT, resulting in less transplantation-related complications. Moreover, clearance of glycosaminoglycans (GAG's) from the bone marrow might improve engraftment. Twenty-two patients with MPS 1H received one or more HCT procedures in combination with ERT. One patient with severe cardiomyopathy improved significantly after ERT. All children were in a relatively good clinical condition before HCT. Of patients 59, 82 and 86% were alive and engrafted after one, two and three HCT procedures, respectively. Two patients died after repetitive HCT. No serious ERT-infusion-related toxicity occurred. ERT with HCT was well tolerated. Neither a positive nor a negative effect on the number of patients who are alive and engrafted after receiving ERT before HCT as compared to a historic cohort was noted. However, patients in a poor clinical condition before HCT might benefit from ERT. PMID- 16715106 TI - Successful hematopoietic stem cell transplantation in a child with active disseminated Mycobacterium fortuitum infection and interferon-gamma receptor 1 deficiency. PMID- 16715107 TI - A study to determine the safety profile and maximum tolerated dose of micafungin (FK463) in patients undergoing haematopoietic stem cell transplantation. AB - This open-label, dose-escalation study assessed the maximum tolerated dose (MTD) of the new antifungal micafungin in patients undergoing haematopoietic stem cell transplantation (HSCT). Participants received 3, 4, 6 or 8 mg/kg/day micafungin intravenously from 7 days to a maximum of 28 days or until neutropaenia resolved. The MTD was defined as the highest dose not causing the same Grade 3 or 4 adverse event in three or more patients. All 36 participants received >/=8 days treatment for a median of 18 days (range: 8-28); 1 patient withdrew consent and a further 11 discontinued to receive another systemic antifungal agent for a suspected infection. No case of confirmed invasive fungal infection occurred. Adverse events were those expected for patients undergoing HSCT and showed no evidence of dose-related toxicity. Criteria for MTD were not met; no patient had a Grade 3 or 4 adverse event considered causally related to micafungin. Thus, the MTD of micafungin can be inferred to be 8 mg/kg/day or higher. PMID- 16715108 TI - Clinical relevance of quantitative varicella-zoster virus (VZV) DNA detection in plasma after stem cell transplantation. AB - Detection of Varicella-Zoster virus (VZV) DNA in plasma can facilitate the early recognition of complicated VZV-infection in immunocompromised hosts. The correlation of VZV-DNA in plasma with clinical presentations of VZV-infection and subsequent aciclovir treatment in allogeneic stem cell transplant (allo-SCT) recipients was studied. In 81 consecutive VZV-IgG positive allo-SCT recipients, VZV-DNA was measured at regular time points (1, 2 and 4 months) following allo SCT and patient records were screened for VZV-related symptoms and aciclovir treatment. Subsequently, possible VZV-cases were studied in detail for the course of VZV-DNA and treatment effects. During the initial screening, VZV-DNA was detectable in seven patients. The survey of VZV-related symptoms revealed five additional possible VZV-cases. In cases where suitable plasma samples were available (10 out of 12), VZV-DNA was present almost simultaneously with the first clinical manifestations. No evidence of a preceding phase detectable by VZV DNA only could be observed. Treatment with aciclovir was associated with a prompt reduction of VZV-DNA load. Detection of VZV-DNA in plasma in allo-SCT recipients accurately reflected the clinical presentation of VZV-infection and treatment with aciclovir. VZV-DNA detection in plasma of allo-SCT recipients appears clinically relevant as this may support early recognition and therapeutic management of VZV-infections following allo-SCT. PMID- 16715109 TI - Bilateral psoas abscesses post-peripheral blood stem cell transplantation. PMID- 16715110 TI - Multiple myeloma-associated AL amyloidosis: is a distinctive therapeutic approach warranted? AB - The natural history of multiple myeloma (MM) was revolutionized by the introduction of haematopoietic stem cell transplantation to the treatment armamentarium of this disease. Defined subgroups of MM patients (such as the elderly or dialysis-dependent) have required an individualized approach in order to minimize the transplant-related mortality. Little, however, is known about the management of 12-30% of MM patients with coexistent AL amyloidosis as the amyloidopathy is often overlooked and when recognized these patients commonly are excluded from clinical trials. While occult amyloidosis appears to have no impact on the toxicity and outcome of MM patients, the presence of symptomatic amyloidopathy clearly worsens their prognosis. Use of induction chemotherapy drugs that can cause further damage to the heart (Adriamycin), nervous system (Vincristine) or kidneys should be avoided as should lengthy delays in proceeding to autograft. Further, refining the transplant eligibility criteria for this subgroup of patients with co-existent amyloidopathy to include the number of organs involved and the degree of cardiac involvement (NYHA class, Troponins and NT-pro-BNP levels) along with melphalan dose-adjustment will minimize the treatment-related toxicity and mortality and possibly allow a reversal of the organ damage induced by the amyloidogenic light chain. PMID- 16715111 TI - Loss of hepatitis A virus antibodies after bone marrow transplantation. AB - Reimmunization guidelines have recommended the inactivated HAV vaccine for hematopoietic stem cell transplant (HSCT) recipients living in or traveling to areas where hepatitis A is endemic. As a shift from high to medium hepatitis A endemicity has been observed in several countries in Latin America, we conducted a retrospective study to evaluate the prevalence of hepatitis A pre-bone marrow transplant (BMT) and the loss of specific antibodies in consecutive stored serum samples from 77 BMT recipients followed up from 82 to 1530 days. The prevalence of HAV antibodies was 92.2% before BMT. As vaccine was not available in Brazil when the samples were taken, it was assumed that this prevalence reflects natural infection. Survival analysis showed that the probability of becoming seronegative was 4.5% (+/-2.6%), 7.9% (+/-3.4%), 10.1% (+/-4.0%), 23.4% (+/-9.6%) at 1, 2, 3 and 4 years after transplant, respectively. The loss of HAV antibodies was significantly associated with longer follow-up (P=0.0015), younger age (P=0.049) and acute graft-versus-host disease (P=0.035). As most reimmunization protocols start around day +365, in developing countries with similar HAV endemicity, BMT recipients should have serological screening before HAV vaccination and the inactivated vaccine should be advised to those seronegative. PMID- 16715112 TI - The laws covering in vitro fertilization and embryo research in Italy. PMID- 16715113 TI - Severe hypermagnesaemia related to laxative use in acute gastrointestinal graft versus-host disease. PMID- 16715114 TI - Splenic rupture following administration of pegfilgrastim in a patient with multiple myeloma undergoing autologous peripheral blood stem cell transplantation. PMID- 16715115 TI - The levosimendan metabolite OR-1896 elicits vasodilation by activating the K(ATP) and BK(Ca) channels in rat isolated arterioles. AB - 1. We characterized the vasoactive effects of OR-1896, the long-lived metabolite of the inodilator levosimendan, in coronary and skeletal muscle microvessels. 2. The effect of OR-1896 on isolated, pressurized (80 mmHg) rat coronary and gracilis muscle arteriole (approximately 150 microm) diameters was investigated by videomicroscopy. 3. OR-1896 elicited concentration-dependent (1 nM-10 microM) dilations in coronary (maximal dilation: 66+/-6%, relative to that in Ca2+-free solutions; pD2: 7.16+/-0.42) and gracilis muscle arterioles (maximal dilation: 73+/-4%; pD2: 6.71+/-0.42), these dilations proving comparable to those induced by levosimendan (1 nM-10 microM) in coronary (maximal dilation: 83+/-6%; pD2: 7.06+/-0.14) and gracilis muscle arterioles (maximal dilation: 73+/-12%; pD2: 7.05+/-0.1). 4. The maximal dilations in response to OR-1896 were significantly (P<0.05) attenuated by the nonselective K+ channel inhibitor tetraethylammonium (1 mM) in coronary (to 34+/-9%) and gracilis muscle arterioles (to 28+/-6%). 5. Glibenclamide (5 or 10 microM), a selective ATP-sensitive K+ channel (KATP) blocker, elicited a greater reduction of OR-1896-induced dilations in skeletal muscle arterioles than in coronary microvessels. 6. Conversely, the selective inhibition of the large conductance Ca2+-activated K+ channels (BK(Ca)) with iberiotoxin (100 nM) significantly reduced the OR-1896-induced maximal dilation in coronary arterioles (to 21+/-6%), but was ineffective in skeletal muscle arterioles (72+/-8%). 7. Accordingly, OR-1896 elicits a substantial vasodilation in coronary and skeletal muscle arterioles, by activating primarily BK(Ca) and K(ATP) channels, respectively, and it is suggested that OR-1896 contributes to the long-term hemodynamic effects of levosimendan. PMID- 16715117 TI - Modulation of P-glycoprotein activity by cannabinoid molecules in HK-2 renal cells. AB - 1. Endogenous and synthetic cannabinoid molecules have been investigated as possible MDR-1/P-glycoprotein (P-gp) modulators in HK-2-immortalized renal cells, using calcein acetoxymethylester (calcein-AM) as a P-gp substrate. 2. Among the endocannabinoid molecules tested, anandamide (AEA), but not 2-arachidonoyl glycerol (2-AG) or palmitoyl-ethanolamide (PEA), increased the intracellular fluorescence emitted by calcein, a metabolic derivative of the P-gp substrate calcein-AM, indicative of a reduction in transport capacity. 3. All the three synthetic cannabimimetics tested, that is, R-(+)-methanandamide (R(+)-MET), AM 251 and CP55,940 significantly increased calcein accumulation in the cytosol. 4. RT-PCR demonstrated that HK-2 cells do not express CB1 or CB2 cannabinoid receptors. 5. R(+)-MET, AM251 and CP55,940 were also evaluated as modulators of P gp expression, by Western blot analysis. Only AM251 weakly enhanced the protein levels (by 1.2-fold) after a 4-day-long incubation with the noncytotoxic drug concentration 2 microM. 6. The present data provide the first evidence that the endocannabinoid AEA and different synthetic cannabinoids may inhibit the P-gp activity in vitro via a cannabinoid receptor-independent mechanism. PMID- 16715116 TI - Role of reactive oxygen species and gp91phox in endothelial dysfunction of pulmonary arteries induced by chronic hypoxia. AB - 1. This study investigates the role of nitric oxide (NO) and reactive oxygen species (ROS) on endothelial function of pulmonary arteries in a mice model of hypoxia-induced pulmonary hypertension. 2. In pulmonary arteries from control mice, the NO-synthase inhibitor Nomega-nitro-L-arginine methyl ester (L-NAME) potentiated contraction to prostaglandin F2alpha (PGF2alpha) and completely abolished relaxation to acetylcholine. In extrapulmonary but not intrapulmonary arteries, acetylcholine-induced relaxation was slightly inhibited by polyethyleneglycol-superoxide dismutase (PEG-SOD) or catalase. 3. In pulmonary arteries from hypoxic mice, ROS levels (evaluated using dihydroethidium staining) were higher than in controls. In these arteries, relaxation to acetylcholine (but not to sodium nitroprusside) was markedly diminished. L-NAME abolished relaxation to acetylcholine, but failed to potentiate PGF2-induced contraction. PEG-SOD or catalase blunted residual relaxation to acetylcholine in extrapulmonary arteries, but did not modify it in intrapulmonary arteries. Hydrogen peroxide elicited comparable (L-NAME-insensitive) relaxations in extra- and intrapulmonary arteries from hypoxic mice. 4. Exposure of gp91phox(-/-) mice to chronic hypoxia also decreased the relaxant effect of acetylcholine in extrapulmonary arteries. However, in intrapulmonary arteries from hypoxic gp91phox(-/-) mice, the effect of acetylcholine was similar to that obtained in mice not exposed to hypoxia. 5. Chronic hypoxia increases ROS levels and impairs endothelial NO-dependent relaxation in mice pulmonary arteries. Mechanisms underlying hypoxia-induced endothelial dysfunction differ along pulmonary arterial bed. In extrapulmonary arteries from hypoxic mice, endothelium-dependent relaxation appears to be mediated by ROS, in a gp91phox-independent manner. In intrapulmonary arteries, endothelial dysfunction depends on gp91phox, the latter being rather the trigger than the mediator of impaired endothelial NO-dependent relaxation PMID- 16715119 TI - Insensitivity of cardiac delayed-rectifier I(Kr) to tyrosine phosphorylation inhibitors and stimulators. AB - 1. The rapidly activating delayed-rectifying K+ current (I(Kr)) in heart cells is an important determinant of repolarisation, and decreases in its density are implicated in acquired and inherited long QT syndromes. The objective of the present study on I(Kr) in guinea-pig ventricular myocytes was to evaluate whether the current is acutely regulated by tyrosine phosphorylation. 2. Myocytes configured for ruptured-patch or perforated-patch voltage-clamp were depolarised with 200-ms steps to 0 mV for measurement of I(Kr) tail amplitude on repolarisations to -40 mV. 3. I(Kr) in both ruptured-patch and perforated-patch myocytes was only moderately (14-20%) decreased by 100 microM concentrations of protein tyrosine kinase (PTK) inhibitors tyrphostin A23, tyrphostin A25, and genistein. However, similar-sized decreases were induced by PTK-inactive analogues tyrphostin A1 and daidzein, suggesting that they were unrelated to inhibition of PTK. 4. Ruptured-patch and perforated-patch myocytes were also treated with promoters of tyrosine phosphorylation, including phosphotyrosyl phosphatase (PTP) inhibitor orthovanadate, exogenous c-Src PTK, and four receptor PTK activators (insulin, insulin-like growth factor-1, epidermal growth factor, and basic fibroblast growth factor). None of these treatments had a significant effect on the amplitude of I(Kr). 5. We conclude that Kr channels in guinea-pig ventricular myocytes are unlikely to be regulated by PTK and PTP. PMID- 16715118 TI - Peptides modified by myristoylation activate eNOS in endothelial cells through Akt phosphorylation. AB - 1. Myristoylated pseudosubstrate of PKCzeta (mPS) - a synthetic myristoylated peptide with a sequence (13 amino acids) mimicking the endogenous PKCzeta pseudosubstrate region -- is considered a selective cell-permeable inhibitor of PKCzeta. We present strong evidence that in endothelial cells the action of mPS is not limited to inhibition of PKC activity and that myristoylation of certain peptides can activate eNOS (endothelial nitric oxide synthase) through Akt phosphorylation. 2. mPS at micromolar concentrations (1-10 microM) induced profound phosphorylation of eNOS, Akt, ERK 1/2, and p38 MAPK in cultured pulmonary artery endothelial cells (PAEC). The same changes were observed after treatment of PAEC with a myristoylated scrambled version of mPS (mScr), whereas a cell-permeable version of PKCzeta pseudosubstrate fused to the HIV-TAT membrane translocating peptide did not induce analogous changes, suggesting that myristoylation confers new properties on the peptides consisting of activation of different signaling pathways in endothelial cells. 3. In addition to mPS and mScr, a number of other myristoylated peptides induced phosphorylation of eNOS suggesting that myristoylation of peptides can activate eNOS by mechanisms unrelated to inhibition of PKC. All active myristoylated peptides contained basic amino acids motif and were longer than six amino acids. 4. Activation of eNOS by myristoylated peptides was dependent on the PI3K/Akt pathway and the rise of intracellular calcium and was associated with an elevation of cGMP levels in PAEC and with relaxation of precontracted isolated pulmonary artery segments. 5. Myristoylated peptides can be considered a new class of activators of NO production in endothelial cells and that using mPS as a specific inhibitor of PKC should be done with caution, especially in endothelial cells. PMID- 16715120 TI - Neuropeptide Y2 receptors are involved in enhanced neurogenic vasoconstriction in spontaneously hypertensive rats. AB - 1. The present study addressed the role of neuropeptide (NPY) Y2 receptors in neurogenic contraction of mesenteric resistance arteries from female spontaneously hypertensive rats (SHR). Arteries were suspended in microvascular myographs, electrical field stimulation (EFS) was performed, and protein evaluated by Western blotting and immunohistochemistry. 2. In vasopressin activated endothelium-intact arteries, NPY and fragments with selectivity for Y1 receptors, [Leu31,Pro34]NPY, Y2 receptors, NPY(13-36), and rat pancreatic polypeptide evoked more pronounced contractions in segments from SHR than in Wistar Kyoto (WKY) arteries, even in the presence of the Y1 receptor antagonist, BIBP3226 (0.3 microM, (R)-N(2)-(diphenacetyl)-N-[(4-hydroxyphenyl)methyl]D arginineamide). 3. In the presence of prazosin and during vasopressin activation, EFS-evoked contractions were larger in arteries from SHR compared to WKY. EFS contractions were enhanced by the Y2 receptor selective antagonist BIIE0246TF (0.5 microM, (S)-N2-[[1-[2-[4-[(R,S)-5,11-dihydro-6(6h)-oxodibenz[b,e]azepin-11 y1]-1-piperazinyl]-2-oxoethyl]cyclo-pentyl-N-[2-[1,2-dihydro-3,5 (4H)-dioxo-1,2 diphenyl-3H-1,2,4-triazol-4-yl]ethyl]-argininamide), reduced by BIBP3226, and abolished by the combination of BIBP3226 and BIIE0246TF. 4. Immunoblotting showed NPY Y1 and Y2 receptor expression to be similar in arteries from WKY and SHR, although a specific Y2 receptor band at 80 kDa was detected only in arteries from WKY. 5. Immunoreaction for NPY was enhanced in arteries from SHR. In contrast to arteries from WKY, BIIE0246TF increased NPY immunoreactivity in EFS-stimulated arteries from SHR. 6. The present results suggest that postjunctional neuropeptide Y1 and Y2 receptors contribute to neurogenic contraction of mesenteric small arteries. Moreover, both enhanced NPY content and altered neuropeptide Y1 and Y2 receptor activation apparently contribute to the enhanced neurogenic contraction of arteries from SHR. PMID- 16715122 TI - Detection of multiple H3 receptor affinity states utilizing [3H]A-349821, a novel, selective, non-imidazole histamine H3 receptor inverse agonist radioligand. AB - 1. A-349821 is a selective histamine H3 receptor antagonist/inverse agonist. Herein, binding of the novel non-imidazole H3 receptor radioligand [3H]A-349821 to membranes expressing native or recombinant H3 receptors from rat or human sources was characterized and compared with the binding of the agonist [3H]N- methylhistamine ([3H]NMH). 2. [3H]A-349821 bound with high affinity and specificity to an apparent single class of saturable sites and recognized human H3 receptors with 10-fold higher affinity compared to rat H3 receptors. [3H]A 349821 detected larger populations of receptors compared to [3H]NMH. 3. Displacement of [3H]A-349821 binding by H3 receptor antagonists/inverse agonists was monophasic, suggesting recognition of a single binding site, while that of H3 receptor agonists was biphasic, suggesting recognition of both high- and low affinity H3 receptor sites. 4. pKi values of high-affinity binding sites for H3 receptor competitors utilizing [3H]A-349821 were highly correlated with pKi values obtained with [3H]NalphaMH, consistent with labelling of H3 receptors by [3H]A-349821. 5. Unlike assays utilizing [3H]NMH, addition of GDP had no effect on saturation parameters measured with [3H]A-349821, while displacement of [3H]A 349821 binding by the H3 receptor agonist histamine was sensitive to GDP. 6. In conclusion, [3H]A-349821 labels interconvertible high- and low-affinity states of the H3 receptor, and displays improved selectivity over imidazole-containing H3 receptor antagonist radioligands. [3H]A-349821 competition studies showed significant differences in the proportions and potencies of high- and low affinity sites across species, providing new information about the fundamental pharmacological nature of H3 receptors. PMID- 16715121 TI - Adrenomedullin acts via nitric oxide and peroxynitrite to protect against myocardial ischaemia-induced arrhythmias in anaesthetized rats. AB - 1. The overall aim of this study was to determine if adrenomedullin (AM) protects against myocardial ischaemia (MI)-induced arrhythmias via nitric oxide (NO) and peroxynitrite. 2. In sham-operated rats, the effects of in vivo administration of a bolus dose of AM (1 nmol kg-1) was assessed on arterial blood pressure (BP), ex vivo leukocyte reactive oxygen species generation and nitrotyrosine deposition (a marker for peroxynitrite formation) in the coronary endothelium. 3. In pentobarbitone-anaesthetized rats subjected to ligation of the left main coronary artery for 30 min, the effects of a bolus dose of AM (1 nmol kg-1, i.v.; n=19) or saline (n=18) given 5 min pre-occlusion were assessed on the number and incidence of cardiac arrhythmias. In a further series of experiments, some animals received infusions of the NO synthase inhibitor N(G)-nitro-L-arginine (LNNA) (0.5 mg kg-1 min-1) or the peroxynitrite scavenger N-mercaptopropionyl-glycine (MPG) (20 mg kg 1 h-1) before AM. 4. AM treatment significantly reduced mean arterial blood pressure (MABP) and increased ex vivo chemiluminescence (CL) generation from leukocytes in sham-operated animals. AM also enhanced the staining for nitrotyrosine in the endothelium of coronary arteries. 5. AM significantly reduced the number of total ventricular ectopic beats that occurred during ischaemia (from 1185+/-101 to 520+/-74; P<0.05) and the incidences of ventricular fibrillation (from 61 to 26%; P<0.05). AM also induced a significant fall in MABP prior to occlusion. AM-induced cardioprotection was abrogated in animals treated with the NO synthase inhibitor LNNA and the peroxynitrite scavenger MPG. 6. This study has shown that AM exhibits an antiarrhythmic effect through a mechanism that may involve generation of NO and peroxynitrite. PMID- 16715123 TI - Grateloupia longifolia polysaccharide inhibits angiogenesis by downregulating tissue factor expression in HMEC-1 endothelial cells. AB - 1. The antiangiogenic and antitumor properties of Grateloupia longifolia polysaccharide (GLP), a new type of polysaccharide isolated from the marine alga, were investigated with several in vitro and in vivo models. Possible mechanisms underlying its antiangiogenic activity were also assessed. 2. GLP dose dependently inhibited proliferation of human microvascular endothelial cells (HMEC-1) and human umbilical vein endothelial cells (HUVEC), with IC50 values of 0.86 and 0.64 mg ml(-1), respectively. In tube formation and cell migration assays using HMEC-1 cells, noncytotoxic doses of GLP significantly inhibited formation of intact tube networks and reduced the number of migratory cells. Inhibition by GLP was VEGF-independent. 3. In the chick chorioallantoic membrane (CAM) assay, GLP (2.5 microg egg(-1)) reduced new vessel formation compared with the vehicle control. GLP (0.1 mg plug(-1)) also reduced the vessel density in Matrigel plugs implanted in mice. 4. The levels of pan and phosphorylated receptors for VEGF, VEGFR-1 (flt-1) and VEGFR-2 (KDR) were not significantly altered by 5 mg ml(-1) GLP treatment of HMEC-1, although tissue factor (TF) showed significant decreases at both mRNA and protein levels following GLP treatment. 5. In mice bearing sarcoma-180 cells, intravenous administration of GLP (200 mg kg(-1)) decreased tumor weight by 52% without obvious toxicity. Vascular density in sections of the tumor was reduced by 64% after GLP treatment. 6. Collectively, these results indicate that GLP has antitumor properties, associated at least, in part, with the antiangiogenesis induced by downregulation of TF. PMID- 16715124 TI - Ras transformation requires metabolic control by 6-phosphofructo-2-kinase. AB - Neoplastic cells transport large amounts of glucose in order to produce anabolic precursors and energy within the inhospitable environment of a tumor. The ras signaling pathway is activated in several cancers and has been found to stimulate glycolytic flux to lactate. Glycolysis is regulated by ras via the activity of 6 phosphofructo-2-kinase/fructose-2,6-bisphosphatases (PFK2/FBPase), which modulate the intracellular concentration of the allosteric glycolytic activator, fructose 2,6-bisphosphate (F2,6BP). We report herein that sequential immortalization and ras-transformation of mouse fibroblasts or human bronchial epithelial cells paradoxically decreases the intracellular concentration of F2,6BP. This marked reduction in the intracellular concentration of F2,6BP sensitizes transformed cells to the antimetabolic effects of PFK2/FBPase inhibition. Moreover, despite co-expression of all four mRNA species (PFKFB1-4), heterozygotic genomic deletion of the inducible PFKFB3 gene in ras-transformed mouse lung fibroblasts suppresses F2,6BP production, glycolytic flux to lactate, and growth as soft agar colonies or tumors in athymic mice. These data indicate that the PFKFB3 protein product may serve as an essential downstream metabolic mediator of oncogenic ras, and we propose that pharmacologic inhibition of this enzyme should selectively suppress the high rate of glycolysis and growth by cancer cells. PMID- 16715125 TI - Overexpression of aurora kinase A in mouse mammary epithelium induces genetic instability preceding mammary tumor formation. AB - Aurora-A/STK15/BTAK, which encodes a centrosome-associated kinase, is amplified and overexpressed in multiple types of human tumors, including breast cancer. However, the causal relationship between overexpression of Aurora-A and tumorigenesis has not been fully established due to contradictory data obtained from different experimental systems. To investigate this, we generated a mouse strain that carries an MMTV-Aurora-A transgene. We showed that all the MMTV Aurora-A mice displayed enhanced branch morphogenesis in the mammary gland and about 40% developed mammary tumors at 20 months of age. The tumor incidence was significantly increased in a p53(+/-) mutation background with about 70% MMTV Aurora-A;p53(+/-) animals developed tumors at 18 months of age. Of note, overexpression of Aurora-A led to genetic instability, characterized by centrosome amplification, chromosome tetraploidization and premature sister chromatid segregation, at stages prior to tumor formation. Most notably, the severe chromosomal abnormality did not cause cell death owing to the activation of AKT pathway, including elevated levels of phosphorylated AKT and mammalian target of rapamycin, and nuclear accumulation of cyclin D1, which enabled continuous proliferation of the tetraploid cells. These data establish Aurora-A as an oncogene that causes malignant transformation through inducing genetic instability and activating oncogenic pathways such as AKT and its downstream signaling. PMID- 16715126 TI - Overexpression of Sp1 transcription factor induces apoptosis. AB - Transcription factor Sp1 has recently been shown to be overexpressed in a number of human cancers and its overexpression contributes to malignant transformation. Sp1 regulates the expression of a number of genes participating in multiple aspects of tumorigenesis such as angiogenesis, cell growth and apoptosis resistance. To better understand the role of increased Sp1 levels on apoptosis regulation we have used retroviruses to overexpress this protein in haematopoietic Baf-3 cells and in 3T3 fibroblasts. We have also used inducible expression systems to control ectopic Sp1 levels in different cell types. Surprisingly, Sp1 overexpression on its own induces apoptosis in all the cellular models tested. The apoptotic pathways induced by Sp1 overexpression are cell type specific. Finally, using a truncated form of Sp1, we show that Sp1-induced apoptosis requires its DNA-binding domain. Our results highlight that Sp1 levels in untransformed cells must be tightly regulated as Sp1 overexpression leads to the induction of apoptosis. Our results also suggest that cancer cells overexpressing Sp1 can avoid Sp1-induced apoptosis. PMID- 16715127 TI - Targeting survivin via PI3K but not c-akt/PKB by anticancer drugs in immature neutrophils. AB - Myelosuppression is the most common unwanted side effect associated with the administration of anticancer drugs, and infections remain a common cause of death in chemotherapy-treated patients. Several mechanisms of the cytotoxicity of these drugs have been proposed and may synergistically operate in a given cell. Survivin expression has been associated with cancer, but recent reports suggest that this molecule is also expressed in several immature and mature hematopoietic cells. Here, we provide evidence that treatment of immature neutrophils with anticancer drugs reduced endogenous survivin levels causing apoptosis. The anticancer drugs did not directly target survivin, instead they blocked the activity of phosphatidylinositol-3-OH kinase, which regulated survivin expression and apoptosis in these cells. Strikingly, and in contrast to other cells, this pathway did not involve the serine/threonine kinase c-akt/PKB. Moreover, in combination with anticancer drug therapy, rapamycin did not induce increased myelosuppression in an experimental lymphoma mouse model. These data suggest that drugs that block either c-akt/PKB or signaling molecules located distal to c akt/PKB may preferentially induce apoptosis of cancer cells as they exhibit no cytotoxicity for immature neutrophils. PMID- 16715128 TI - Rapamycin inhibits cell motility by suppression of mTOR-mediated S6K1 and 4E-BP1 pathways. AB - Rapamycin, an inhibitor of the mammalian target of rapamycin (mTOR), inhibits tumor cell motility. However, the underlying mechanism is poorly understood. Here, we show that rapamycin inhibited type I insulin-like growth factor (IGF-I) stimulated motility of a panel of cell lines. Expression of a rapamycin-resistant mutant of mTOR (mTORrr) prevented rapamycin inhibition of cell motility. However, cells expressing a kinase-dead mTORrr remained sensitive to rapamycin. Downregulation of raptor or rictor by RNA interference (RNAi) decreased cell motility. However, only downregulation of raptor mimicked the effect of rapamycin, inhibiting phosphorylation of S6 kinase 1 (S6K1) and 4E-BP1. Cells infected with an adenovirus expressing constitutively active and rapamycin resistant mutant of p70 S6K1, but not with an adenovirus expressing wild-type S6K1, or a control virus, conferred to resistance to rapamycin. Further, IGF-I failed to stimulate motility of the cells, in which S6K1 was downregulated by RNAi. Moreover, downregulation of eukaryotic initiation factor 4E (eIF4E)-binding protein 1 (4E-BP1) by RNAi-attenuated rapamycin inhibition of cell motility. In contrast, expression of constitutively active 4E-BP1 dramatically inhibited IGF-I stimulated cell motility. The results indicate that both S6K1 and 4E-BP1 pathways, regulated by TORC1, are required for cell motility. Rapamycin inhibits IGF-I-stimulated cell motility, through suppression of both S6K1 and 4E-BP1/eIF4E signaling pathways, as a consequence of inhibition of mTOR kinase activity. PMID- 16715129 TI - Identification of target genes in laryngeal squamous cell carcinoma by high resolution copy number and gene expression microarray analyses. AB - Molecular mechanisms contributing to initiation and progression of head and neck squamous cell carcinoma are still poorly known. Numerous genetic alterations have been described, but molecular consequences of such alterations in most cases remain unclear. Here, we performed an integrated high-resolution microarray analysis of gene copy number and expression in 20 laryngeal cancer cell lines and primary tumors. Our aim was to identify genetic alterations that play a key role in disease pathogenesis and pinpoint genes whose expression is directly impacted by these events. Integration of DNA level data from array-based comparative genomic hybridization with RNA level information from oligonucleotide microarrays was achieved with custom-developed bioinformatic methods. High-level amplifications had a clear impact on gene expression. Across the genome, overexpression of 739 genes could be attributed to gene amplification events in cell lines, with 325 genes showing the same phenomenon in primary tumors including FADD and PPFIA1 at 11q13. The analysis of gene ontology and pathway distributions further pinpointed genes that may identify potential targets of therapeutic intervention. Our data highlight genes that may be critically important to laryngeal cancer progression and offer potential therapeutic targets. PMID- 16715130 TI - A ubiquitin ligase, skeletrophin, is a negative regulator of melanoma invasion. AB - Skeletrophin (mindbomb homolog 2 (MIB2)) is a RING (Really Interesting New Gene) finger-dependent ubiquitin ligase, which targets the intracellular region of Notch ligands. A previous immunohistochemical study demonstrated that skeletrophin was downregulated in many melanomas. In the present study, we have identified a promoter region of skeletrophin on a CpG island and detected aberrant methylation of this region in six of 31 invasive melanomas, but in none of 25 benign nevi or five non-invasive superficial spreading melanomas. Subsequently, we found that a zinc-finger transcriptional factor Snail, which is overexpressed in many melanoma cells, repressed the skeletrophin promoter activity via an E-box-related element and was involved in downregulation of skeletrophin. An activator protein-2, which has a tumor suppressor-like role in melanoma, increased skeletrophin expression. Interestingly, exogenously expressed skeletrophin reduced melanoma cell invasion in vitro and in vivo. Colony formation in soft agar was also reduced in a RING motif-dependent manner, without affecting cell growth. We also found that skeletrophin downregulated transcription of the Met oncogene, which encodes the hepatocyte growth factor receptor and plays a role in the determination of the invasive phenotype of many malignant tumors. Finally, exogenously expressed skeletrophin, but not its RING mutant, increased transcription of Hes1 gene, a downstream effector of Notch pathway in melanoma cells. The present findings indicate that skeletrophin might be a novel suppressor factor for melanoma invasion. PMID- 16715131 TI - Acetylcholinesterase expression mediated by c-Jun-NH2-terminal kinase pathway during anticancer drug-induced apoptosis. AB - It has been shown that acetylcholinesterase (AChE) expression was induced during apoptosis and the anti-sense oligonucleotides and siRNA of AChE may prevent apoptosis in various cell types. However, the mechanisms underlying AChE upregulation remain elusive. We demonstrated here that c-Jun NH2-terminal kinase (JNK) could mediate AChE expression. In this study, both etoposide and excisanin A, two anticancer agents, induced apoptosis in colon cancer cell line SW620 as determined by Annexin V staining, the cleavage of caspase-3 and the proteolytic degradation of poly (ADP-ribose) polymerase (PARP). The results showed that both the agents upregulated AChE in SW620 cells. In the meantime, JNK was also activated and the expression and phosphorylation of c-Jun increased in SW620 cells exposed to the two agents. The induced AChE mRNA and protein expression could be blocked by SP600125, a specific inhibitor of SAPK/JNK, and small interfering RNA directed against JNK1/2. Transfection with adenovirus-mediated dominant negative c-Jun also blocked the upregulation of AChE expression. Together, these results suggest that AChE expression may be mediated by the activation of JNK pathway during apoptosis through a c-Jun-dependent mechanism. PMID- 16715132 TI - HER2 signaling modulates the equilibrium between pro- and antiangiogenic factors via distinct pathways: implications for HER2-targeted antibody therapy. AB - We determined the impact of HER2 signaling on two proangiogenic factors, vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8), and on an antiangiogenic factor, thrombospondin-1 (TSP-1). Re-expression of HER2 in MCF-7 and T-47D breast cancer cells that endogenously express low levels of HER2 resulted in elevated expression of VEGF and IL-8 and decreased expression of TSP 1. Inhibition of HER2 with a humanized anti-HER2 antibody (trastuzumab, or Herceptin) or a retrovirus-mediated small interfering RNA against HER2 (siHER2) decreased VEGF and IL-8 expression, but increased TSP-1 expression in BT474 breast cancer cells that express high levels of HER2. These in vitro results were further evaluated by treatment of BT474 xenografts in immunosuppressed mice with trastuzumab. Trastuzumab inhibited growth of BT474 xenografts and decreased microvascular density associated with downregulation of VEGF and IL-8 and with upregulation of TSP-1 expression. Inhibiting the PI3K-AKT pathway decreased VEGF and IL-8 expression. AKT1 overexpession increased VEGF and IL-8 expression, but did not increase TSP-1 expression. A p38 kinase inhibitor, SB203580, instead blocked TSP-1 expression and a p38 activator, MKK6, increased TSP-1 expression. Trastuzumab stimulated sustained p38 activation and SB203580 attenuated the TSP-1 upregulation induced by trastuzumab. HER2 signaling therefore influences the equilibrium between pro- and antiangiogenic factors via distinct signaling pathways. Trastuzumab inhibits angiogenesis and tumor growth, at least in part, through activation of the HER2-p38-TSP-1 pathway and inhibition of the HER2-PI3K AKT-VEGF/IL-8 pathway. PMID- 16715133 TI - Dual role of TMS1/ASC in death receptor signaling. AB - Aberrant DNA methylation of promoter region CpG islands is associated with gene silencing and serves as an alternative to mutations in the inactivation of tumor suppressor genes in human cancers. We identified a gene TMS1 (for Target of Methylation-mediated Silencing) that is subject to such epigenetic silencing in a significant proportion of human breast and other cancers. Also known as ASC and PYCARD, TMS1 encodes a bipartite intracellular signaling molecule with proposed roles in apoptosis and inflammation. However, the precise role of this protein in the pathogenesis of breast and other cancers has not been clearly defined. In this study, we examined the role of TMS1/ASC in death receptor signaling. We found that TMS1/ASC is upregulated in response to treatment with TNF-related apoptosis-inducing ligand (TRAIL) and tumor necrosis factor-alpha (TNFalpha) in breast epithelial cells, but not in human fibroblasts. This upregulation was not dependent on the synthesis of a TNFalpha-regulated intermediate or alterations in mRNA stability, suggesting a direct effect on TMS1/ASC transcription. Induction of TMS1/ASC by TNFalpha was blocked by co-expression of a dominant negative IkappaBalpha, small interfering RNA-mediated knockdown of RelA/p65, or concurrent treatment with SP600125, indicating a requirement for the nuclear factor-kappaB (NF-kappaB) and jun kinase signaling pathways. Although previous work has suggested that TMS1/ASC may be directly regulated by p53, we found that whereas treatment of breast epithelial cells or normal diploid fibroblasts with DNA damaging agents resulted in the stabilization of endogenous p53 and a concomitant increase in p21, it had little impact on the expression of TMS1/ASC mRNA or protein. We further show that whereas TMS1/ASC is not required for TNFalpha or TRAIL-induced activation of NF-kappaB or caspase-8, it can promote caspase-8 activation independently of death receptor-ligand interactions. Taken together, these data suggest that upregulation of TMS1/ASC by TNFalpha and subsequent activation of caspase-8 could function to amplify the apoptotic signal induced by death receptors in some cell types, including breast epithelial cells. PMID- 16715134 TI - Reciprocal regulation of RhoA and RhoC characterizes the EMT and identifies RhoC as a prognostic marker of colon carcinoma. AB - Understanding how RhoC expression and activation are regulated is essential for deciphering its contribution to tumorigenesis. Here, we report that RhoC expression and activation are induced by the epithelial to mesenchymal transition (EMT) of colon carcinoma. Using LIM 1863 colon cancer cells, RhoC protein expression and subsequent activation were detected coincident with the loss of E cadherin and acquisition of mesenchymal characteristics. Several Ets-1 binding sites were identified in the RhoC promoter, and evidence was obtained using chromatin immunoprecipitation that Ets-1 can regulate RhoC expression during the EMT. Interestingly, a marked decrease in RhoA activation associated with the EMT was observed that corresponds to the increase in RhoC expression. Use of shRNA established that RhoA inhibits and RhoC promotes post-EMT cell migration, demonstrating functional significance for their coordinate regulation. To assess the importance of RhoC expression in colon cancer, immunohistochemistry was performed on 566 colorectal tumors with known clinical outcome. The level of RhoC ranged from no expression to high expression, and statistical analysis revealed that elevated RhoC expression correlates with poor outcome as well as aberrant expression and localization of E-cadherin. These data provide one mechanism for how RhoC expression is regulated in colon carcinoma and substantiate its utility as a prognostic marker. PMID- 16715135 TI - Epigenetic regulation of X-linked cancer/germline antigen genes by DNMT1 and DNMT3b. AB - We examined the function of two key DNA methyltransferase (DNMT) enzymes in epigenetic regulation of X-linked cancer/germline (CG-X) antigen genes in human cancer cells, using MAGE-A1, NY-ESO-1, and XAGE-1 as models. In HCT116 cells, genetic knockout of DNMT1 caused moderate activation of CG-X genes, DNMT3b knockout had a negligible effect, and double knockout of both enzymes caused robust gene induction. Similarly, dual DNMT knockout caused dramatic hypomethylation of the MAGE-A1 and NY-ESO-1 promoters, DNMT1 knockout showed moderate hypomethylation, and DNMT3b knockout elicited only slight methylation changes. In contrast, both single and double knockout cells showed significant hypomethylation of the XAGE-1 promoter. RNA interference (RNAi) targeting of DNMT1 in HCT116 cells validated the results seen using genetic knockout cells; however, RNAi targeting of DNMT1 in a different colorectal cancer cell line revealed a greater independent role for DNMT1 in mediating CG-X gene repression and promoter methylation in other cell types. Notably, the histone H3 modification pattern at CG-X promoters was altered following DNMT knockout. DNMT1 or DNMT3b knockout reduced dimethylated lysine-9 (diMe-H3K9) levels, but did not significantly affect dimethylated lysine-4 (diMe-H3K4) or acetylated lysine-9 (Ac H3-K9) levels. In contrast, dual DNMT1/3b knockout reduced the level of diMe-H3K9 and dramatically increased the levels of diMe-H3K4 and Ac-H3K9 at CG-X gene loci. In summary, DNMT1 and DNMT3b were found to perform both redundant and independent functions in epigenetic regulation of CG-X antigen genes in human cancer cells. PMID- 16715137 TI - Human melanoma cells expressing V600E B-RAF are susceptible to IGF1R targeting by small interfering RNAs. AB - The type 1 insulin-like growth factor receptor (IGF1R) is overexpressed by malignant melanomas compared with benign naevi, and mediates proliferation, motility and protection from apoptosis. However, the utility of IGF1R targeting as anti-cancer therapy may be limited by activating mutations in downstream signaling intermediates. We previously showed that IGF1R knockdown blocked survival of prostate cancer cells in which Akt activation was deregulated by PTEN loss. The current study investigated effects of IGF1R targeting in cells harboring activating RAS-RAF mutations, found in 70-80% of human melanomas. We assembled a panel of eight human melanoma cell lines: two expressing wild-type (WT) B-RAF and N-RAS, two with activating N-RAS mutations and four harboring V600E B-RAF. We also generated isogenic cell populations overexpressing WT or V600E B-RAF. Cells expressing V600E B-RAF were relatively resistant to apoptosis. However, IGF1R gene silencing was capable of inducing significant inhibition of survival, enhancement of apoptosis, and approximately two-fold sensitization to cisplatin and temozolomide. These effects were independent of mutation status and were associated with reduced activation of Akt and also, unexpectedly, of ERKs. These results support development of IGF1R targeting as therapy for melanoma, regardless of the presence of activating mutations in the RAS-RAF pathway. PMID- 16715136 TI - Gab1 signaling is regulated by EGF receptor sorting in early endosomes. AB - Although combinatorial signaling through the ErbB network is implicated in certain types of human cancer, the specifics of how particular receptors contribute to the transformed phenotype are not well understood. The goal of this study was to identify epidermal growth factor (EGF) receptor-dependent cell signaling abnormalities specifically associated with mutations in a previously described 679-LL lysosomal sorting signal, which restrict ligand-dependent receptor downregulation by promoting recycling. Importantly, the 679-LL signal is not conserved in any of the other members of the ErbB receptor family suggesting its physiological function may be tightly regulated during EGF receptor-dependent signaling. Our data indicate that cells expressing receptors with an inactive 679 AA signal are rapidly transported to Rab4+ early endosomes after they are internalized in contrast to wild-type receptors that are localized to early endocytic antigen 1 (EEA1)+ early endosomes. Divergent trafficking in early endosomes is associated with prolonged activation of p44/42 mitogen-activated protein kinases (MAPK) but not Akt. Gab1 appears to be the critical signaling molecule facilitating prolonged MAPK signaling, and activated Gab1 is recruited to early endosomes in 679-AA receptor-expressing cells. Activated Gab1 is also recruited to early endosomes in breast cancer cells characterized by high levels of EGF receptor-ErbB2 heterodimers, suggesting 679-AA expressing cells recapitulate certain aspects of EGF receptor signaling and transformation by activated ErbB2. Phosphatidylinositol 3-kinase (PI3K)-dependent membrane translocation known to be important for maintaining Gab1 activity in other settings was dispensable. We conclude that 679-LL has dual functions in EGF receptor trafficking and threshold signaling through a subset of signaling molecules including p44/42 MAPK and Gab1. PMID- 16715138 TI - Identification of secondary structure in the 5'-untranslated region of the human adrenomedullin mRNA with implications for the regulation of mRNA translation. AB - Adrenomedullin (AM) is a multifunctional regulatory peptide with important angiogenic and mitogenic properties. Here we identify a region of stable secondary structure in the 5'-untranslated region (5' UTR) of human AM mRNA. Reverse transcriptase-polymerase chain reaction of the 5' UTR consistently resulted, in addition to the product with the expected size of 155 base pair (bp), in a second product with an approximately 65-bp deletion from the central region of the 5' UTR, suggesting the presence of a secondary structure. The presence of a stem-loop structure was confirmed by probing the 5' UTR with RNases with selectivity for single- or double-stranded RNA. We investigated the role of this stem-loop structure in expression of luciferase reporter gene in cultured cell lines. Reporter assays using a chimeric mRNA that combined luciferase and the 5' UTR of AM mRNA demonstrated a dramatic decrease of the reporter activity owing to a decreased translation, whereas the deletion of the stem-loop structure localized between nt +31 and +95 from the cap site led to the recovery of activity. Gel migration shift assays using cytosolic extracts from mammalian cell lines demonstrate a specific binding of a cytosolic protein to riboprobes containing the 5' UTR of AM but not to riboprobes either corresponding to other areas of the message or containing the 5' UTR but lacking the region of secondary structure. Although we conclude that the 5' UTR of the human AM mRNA can modulate the translation of AM mRNA in vivo, and that the predicted stem-loop structure is necessary for this inhibition, the functional consequences of the cis element binding activity remain to be determined. PMID- 16715139 TI - Evaluation of potential mechanisms underlying genotype-phenotype correlations in multiple endocrine neoplasia type 2. AB - Distinct dominant activating mutations in the RET proto-oncogene are responsible for the development of multiple endocrine neoplasia type 2 (MEN 2). Concise examination of the mutated codons led to the detection of a striking genotype phenotype correlation between the mutated codon and the MEN 2 phenotype in terms of onset and aggressiveness of the disease, suggesting that manifestation and clinical progression is conditioned by the type of mutation. To gain insight into the molecular basis for this genotype-phenotype correlation, we analysed the impact of common and rare mutations identified in MEN 2A (C609Y, C634R), MEN 2B (A883F, M918T) and familial medullary thyroid carcinoma (Y791F) patients on several aspects of cell transformation, including proliferation, apoptosis, anchorage-independent growth and signaling. We found that tumor cells arising from distinct extracellular or intracellular MEN 2 mutations clearly differ in their proliferation properties owing to the activation of different molecular pathways, but importantly, also in resistance to apoptosis. Whereas MEN 2A mutants resulted in accelerated cell proliferation, MEN 2B-RET mutants significantly enhanced suppression of apoptosis, which may account, at least partially, for some of the clinical differences in MEN 2 patients. PMID- 16715140 TI - Endogenous production of leukotriene D4 mediates autocrine survival and proliferation via CysLT1 receptor signalling in intestinal epithelial cells. AB - The cysteinyl leukotriene1 (CysLT1) receptor (CysLT1R) enhances survival and proliferation of intestinal cells via distinct pathways. Here, we have demonstrated that there is significant endogenous production of CysLTs from both non-tumour- and tumour-derived intestinal epithelial cells. Treatment of two non tumour cell lines, Int 407 and IEC-6, with CysLT1R antagonists led to shrinkage and detachment of cells, confirmed as apoptotic cell death, and a dose-dependent reduction in proliferation. However, in the tumour intestinal cell lines Caco-2, SW480, HCT-116 and HT-29, treatment with CysLT1R antagonists significantly reduced proliferation, but had no effect on apoptosis. A unique characteristic of intestinal cancer cells is the presence of nuclear CysLT1Rs, which are inaccessible to receptor antagonists. In these cells, inhibition of the endogenous production of CysLTs indirectly, by 5-lipoxygenase inhibition, impaired CysLT1R signalling throughout the cell, and resulted in apoptosis of the tumour cells. These data reveal the existence of constitutive CysLT1R signalling that mediates both survival and proliferation in intestinal cells. Importantly, we propose that tumour-derived intestinal cells are resistant to CysLT1R antagonist-induced apoptosis, a phenomena that could be explained by nuclear CysLT1R signalling. PMID- 16715141 TI - Induction of the adenoma-carcinoma progression and Cdc25A-B phosphatases by the trefoil factor TFF1 in human colon epithelial cells. AB - TFF1 is overexpressed in inflammatory diseases and human cancers of the digestive and urogenital systems. To examine the transforming potential of TFF1 in human colon epithelial cells, premalignant PC/AA/C1 adenoma cells (PC) derived from a patient with familial adenomatous polyposis (FAP) were transformed by the TFF1 cDNA and used as a model of the adenoma-carcinoma transition. Constitutive expression of TFF1 increased anchorage-independent cell growth in soft agar, and induced or potentiated the growth of colon PC-TFF1 and kidney MDCKts.src-TFF1 tumor xenografts in athymic mice. This resulted in reduction of thapsigargin induced apoptosis and promotion of collagen type I invasion through several oncogenic pathways. Using the differential display approach to identify TFF1 target genes, we found that the dual specific phosphatases Cdc25A and B implicated in cell cycle transitions are strongly upregulated under active forms in both PC-TFF1 and HCT8/S11-TFF1 colon cancer cells. Accordingly, TFF1 expression is absent in normal human colon crypts but is induced in correlation with Cdc25a and b transcript levels and tumor grade in familial and sporadic colon adenomas and carcinomas. We propose that TFF1 and Cdc25A-B cooperate with other dominant oncogenic pathways to induce the adenoma and adenocarcinoma transitions. Agents that target TFF1/Cdc25 signaling pathways may be useful for treating patients with TFF1-positive solid tumors. PMID- 16715142 TI - Regulation of transendothelial migration of colon cancer cells by E-selectin mediated activation of p38 and ERK MAP kinases. AB - The invasive properties of cancer cells depend on their intrinsic motile potential and on their ability to breach the endothelial barrier. In the present work, we investigated the mechanisms by which adhesion of colon cancer cells to E selectin expressed by endothelial cells regulates the barrier function of these cells and modulates transmigration of cancer cells. We found that the stimulation of E-selectin by activating antibodies or the adhesion of HT-29 cells results in an increase in the activity of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinases. In turn, the activation of p38 and ERK enhances transendothelial permeability and migration of HT-29 cells. We also obtained evidence suggesting that p38-mediated increase in transendothelial migration of cancer cells depends on a myosin light chain phosphorylation mediated formation of stress fibres. On the other hand, the activation of ERK by E-selectin modulates the opening of interendothelial spaces by initiating the activation of Src kinase activities and the dissociation of the VE-cadherin/beta catenin complex. We conclude that activation of E-selectin by adhering cancer cells is an important process that regulates the extravasation of colon cancer cells by initiating p38- and ERK-dependent mechanisms that both contribute to regulate the integrity of the endothelial layer. PMID- 16715143 TI - Genomic loss and epigenetic silencing of very-low-density lipoprotein receptor involved in gastric carcinogenesis. AB - Homozygous loss in the genomic sequence, a mechanism for inactivating tumor suppressor genes (TSGs) in cancer, has been used as a tag for the identification of novel TSGs, and array-based comparative genomic hybridization (array-CGH) has a great potential for high-throughput identification of this change. We identified a homozygous loss of the very-low-density lipoprotein receptor (VLDLR) gene (9p24.2) from genome-wide screening for copy-number alterations in 32 gastric cancer (GC) cell lines using array-CGH. Although previous reports demonstrated mRNA or protein expression of VLDLR in various cancers including GC, the association between genomic losses or epigenetic silencing of this gene and carcinogenesis has never been reported before. Homozygous deletion of VLDLR was also seen in primary GCs, albeit infrequently, and about half of GC cell lines showed lost or reduced VLDLR expression. The VLDLR expression was restored in gene-silenced GC cells after treatment with 5-aza 2'-deoxycytidine. According to methylation analyses, hypermethylation of the VLDLR promoter region, which all of GC lines without its expression showed, occurred in some primary GCs. Restoration of VLDLR type I expression in GC cells reduced colony formation. These results suggest that not only the expression of VLDLR but also genetic or epigenetic silencing of this gene may contribute to tumor formation and be involved in gastric carcinogenesis. PMID- 16715144 TI - Factorial coding of natural images: how effective are linear models in removing higher-order dependencies? AB - The performance of unsupervised learning models for natural images is evaluated quantitatively by means of information theory. We estimate the gain in statistical independence (the multi-information reduction) achieved with independent component analysis (ICA), principal component analysis (PCA), zero phase whitening, and predictive coding. Predictive coding is translated into the transform coding framework, where it can be characterized by the constraint of a triangular filter matrix. A randomly sampled whitening basis and the Haar wavelet are included in the comparison as well. The comparison of all these methods is carried out for different patch sizes, ranging from 2x2 to 16x16 pixels. In spite of large differences in the shape of the basis functions, we find only small differences in the multi-information between all decorrelation transforms (5% or less) for all patch sizes. Among the second-order methods, PCA is optimal for small patch sizes and predictive coding performs best for large patch sizes. The extra gain achieved with ICA is always less than 2%. In conclusion, the edge filters found with ICA lead to only a surprisingly small improvement in terms of its actual objective. PMID- 16715145 TI - Scene estimation from speckled synthetic aperture radar imagery: Markov-random field approach. AB - A novel Markov-random-field model for speckled synthetic aperture radar (SAR) imagery is derived according to the physical, spatial statistical properties of speckle noise in coherent imaging. A convex Gibbs energy function for speckled images is derived and utilized to perform speckle-compensating image estimation. The image estimation is formed by computing the conditional expectation of the noisy image at each pixel given its neighbors, which is further expressed in terms of the derived Gibbs energy function. The efficacy of the proposed technique, in terms of reducing speckle noise while preserving spatial resolution, is studied by using both real and simulated SAR imagery. Using a number of commonly used metrics, the performance of the proposed technique is shown to surpass that of existing speckle-noise-filtering methods such as the Gamma MAP, the modified Lee, and the enhanced Frost. PMID- 16715146 TI - Multimodel Kalman filtering for adaptive nonuniformity correction in infrared sensors. AB - We present an adaptive technique for the estimation of nonuniformity parameters of infrared focal-plane arrays that is robust with respect to changes and uncertainties in scene and sensor characteristics. The proposed algorithm is based on using a bank of Kalman filters in parallel. Each filter independently estimates state variables comprising the gain and the bias matrices of the sensor, according to its own dynamic-model parameters. The supervising component of the algorithm then generates the final estimates of the state variables by forming a weighted superposition of all the estimates rendered by each Kalman filter. The weights are computed and updated iteratively, according to the a posteriori-likelihood principle. The performance of the estimator and its ability to compensate for fixed-pattern noise is tested using both simulated and real data obtained from two cameras operating in the mid- and long-wave infrared regime. PMID- 16715147 TI - Iterative image reconstruction using prior knowledge. AB - A method is proposed to reconstruct signals from incomplete data. The method, which can be interpreted both as a discrete implementation of the so-called prior discrete Fourier transform (PDFT) spectral estimation technique and as a variant of the algebraic reconstruction technique, allows one to incorporate prior information about the reconstructed signal to improve the resolution of the signal estimated. The context of diffraction tomography and image reconstruction from samples of the far-field scattering amplitude are used to explore the performance of the method. On the basis of numerical computations, the optimum choice of parameters is determined empirically by comparing image reconstructions of the noniterative PDFT algorithm and the proposed iterative scheme. PMID- 16715148 TI - Optical superresolution of focused partially spatially coherent laser beams. AB - We report design theories of a diffractive superresolution element (DSE) to implement optical superresolution of focused partially spatially coherent laser beams. The design problem of the DSE can be transformed into a problem of linear programming to obtain a globally optimal solution. By using the design theories, some fundamental limits of optical superresolution of focused partially spatially coherent laser beams are proposed, and several design examples are provided. As expected, both the fundamental limits and the design examples show that worse spatial coherence will cause worse superresolution performance. The design theories provide a design approach with partially coherent beams and may be useful for other design problems under partially coherent illumination. PMID- 16715149 TI - Evaluation study of the effectiveness of the integrated genetic-algorithm-based strategy for the tomographic subsurface detection of defects. AB - An assessment is presented of the integrated genetic-algorithm strategy based on a numerically computed Green's function for subsurface inverse scattering problems arising in nondestructive evaluation/testing industrial applications. To show the effectiveness and current limitations of such a microwave technique in dealing with various scenarios characterized by lossless and lossy host media as well as in noisy environments, several numerical experiments are considered. The results obtained confirm the effectiveness of the approach in fully exploiting the available a priori information through a suitable scattering model, which allows a nonnegligible enhancement of the reconstruction accuracy as well as a reduction of the overall computational burden with respect to standard imaging approaches. PMID- 16715150 TI - Numerical calculation of a converging vector electromagnetic wave diffracted by an aperture using Borgnis potentials. II. Application to the study of focal shift. AB - Focal shift of the converging spherical wavefront light diffracted by a circular aperture is numerically studied with the method of calculating the vector diffractive field by using Borgnis potentials given in Part I [J. Opt. Soc. Am. A23, 872 (2006)]. The quantitative dependence of the focal shift on the geometric parameters is discussed. The focal shift is mainly determined by the Fresnel number (N(f)) on the geometric focusing plane of the converging light, and an empirical formula between the fractional focal shift and the Fresnel number is deduced for N(f)<2. The focal shift of the same geometry is also studied on the basis of the scalar Rayleigh theory of diffraction, and its comparison with and difference from the result of our method are presented. PMID- 16715151 TI - Metal iris influence on guided-mode diffraction. AB - The effects of the presence of metal irises on guided-mode propagation through a symmetrical three-layer slab waveguide are examined by using the integral equation method. The aperture electric field distribution is expressed in terms of a finite series of Chebyshev polynomials. The modal reflection and transmission coefficients, the near-field structure, and the far-field radiation pattern are calculated, while numerical results are presented for several iris apertures. PMID- 16715152 TI - Partially coherent analysis of imaging and interferometric phased arrays: noise, correlations, and fluctuations. AB - Phased arrays are of considerable importance for far-infrared, submillimeter wave, and microwave astronomy; they are also being developed for areas as diverse as optical switching, radar, and radio communications. We present a discretized, modal theory of imaging and interferometric phased arrays. It is shown that the average powers, field correlations, power fluctuations, and correlations between power fluctuations at the output ports of an imaging, or interferometric, phased array can be determined for a source in any state of spatial coherence and polarization, once the synthesized beam patterns are known. It is not necessary to know anything about the internal construction of the beam-forming networks; indeed, the beam patterns can be taken from experimental data. The synthesized beams can be nonorthogonal and even linearly dependent. Our theory leads to many conceptual insights and opens the way to a range of new design and simulation techniques. PMID- 16715153 TI - Spatial coherence effects in light scattering from metallic nanocylinders. AB - We study the scattering of a partially coherent electromagnetic beam from metallic nanocylinders and analyze the effects of plasmon resonances on the coherence and polarization properties of the optical near field. We employ the coherent-mode representation for the incident field and solve the scattering problem independently for each mode by using a boundary-integral method. Our results show that the plasmon resonances may significantly affect the coherence and polarization characteristics of the near field and that partial coherence influences the energy flow in nanocylinder arrays. PMID- 16715154 TI - Multispectral intensity diffraction tomography reconstruction theory: quasi nondispersive objects. AB - A multispectral intensity diffraction tomography (I-DT) reconstruction theory for quasi-nondispersive scattering objects is developed and investigated. By "quasi nondispersive" we refer to an object that is characterized by a refractive index distribution that is approximately nondispersive over a predefined finite temporal frequency interval in which the tomographic measurements are acquired. The scanning requirements and measurement data are shown to be different than in conventional I-DT. Unlike conventional I-DT that requires intensity measurements on a pair of detector planes for each probing wave field, this new method uses measurements on a single detector plane at two frequencies. Computer simulation studies are conducted to demonstrate the method. PMID- 16715155 TI - Finite-size effects in surface-enhanced Raman scattering in noble-metal nanoparticles: a semiclassical approach. AB - We study finite-size effects in surface-enhanced Raman scattering (SERS) from molecules adsorbed on small metal particles. Within an electromagnetic description of SERS, the enhancement of the Raman signal originates from the local field of the surface plasmon resonance in a nanoparticle. With decreasing particle sizes, this enhancement is reduced due to the size-dependent Landau damping of the surface plasmon. We show that, in small noble-metal particles, the reduction of interband screening in the surface layer leads to an additional increase in the local field acting on a molecule close to the metal surface. The overall size dependence of Raman signal enhancement is determined by the interplay between Landau damping and underscreening effects. Our calculations, based on a two-region model, show that the role of the surface layer increases for smaller nanoparticle sizes due to a larger volume fraction of the underscreened region. PMID- 16715156 TI - Wavefront correction with high-order curvature adaptive optics systems. AB - We demonstrate the existence of higher-order curvature adaptive optics (AO) systems and compare their performance with the current 85-element system being built at the Institute for Astronomy at the University of Hawaii. Simulation results show that systems with in excess of 500 actuators are possible with actuator patterns that are simple extensions of the 85-element design. The attenuation of residual phase error within the Nyquist frequency of the deformable mirror (DM) satisfies the (-5/6) power law. A high-order system is also analyzed in which the pattern of wavefront sensor is synthesized from square pixels and the curvature actuators of the DM are also rectangular. The Strehl performance is approximately 2% worse than its annular analog. PMID- 16715157 TI - Rapid simulation of steady-state spatially resolved reflectance and transmittance profiles of multilayered turbid materials. AB - We present a technique for efficiently computing the reflection and transmission of light by arbitrary systems of turbid layers. To approximate the steady-state reflectance and transmittance without the need to solve difficult boundary conditions, we convolve the reflectance and transmittance profiles of individual layers. We extend single-slab boundary conditions to handle index-of-refraction mismatches between turbid slabs and account for interlayer scattering by applying methods similar to Kubelka-Munk theory in frequency space. We demonstrate good agreement between the reflectance and the transmittance predicted by our model and numerical Monte Carlo methods and show that the far-source reflectance and transmittance of multilayered turbid materials are dominated by interlayer scattering. PMID- 16715158 TI - Time-domain frequency-selective processing in nuclear magnetic resonance: a spatial-spectral holographic perspective. AB - The fundamental nuclear magnetic resonance (NMR) imaging equation can be derived from a spatial-spectral holographic wavefront reconstruction formulation similar to that in quantum optics. A spatial-spectral holographic interpretation arises naturally in NMR from the inhomogeneous linewidth broadening due to either an imposed set of linear orthogonal gradient fields or from the intrinsic chemical anisotropy of the spin system. We can thus think of NMR k-space as a spatial spectral holographic grating. The spatial holographic component arises from dielectric effects at high field strength (>4 T) when the excitation wavelength is less than or commensurate with the size of the imaging sample. The holographic properties of storage, time-reversal, recognition, and triple correlations are experimentally demonstrated in an inhomogeneously broadened NMR sample. This holographic NMR interpretation has additional implications on selective radio frequency pulse design, microscopy imaging, and the use of conjugate imaging for field inhomogeneity corrections using the time-reversed component of the readout, to be the subject of a subsequent paper. PMID- 16715159 TI - Spiral interferogram analysis. AB - Interference microscopy using spatial Fourier filtering with a vortex phase element leads to interference fringes that are spirals rather than closed rings. Depressions and elevations in the optical thickness of the sample can be distinguished immediately by the sense of rotation of the spirals. This property allows an unambiguous reconstruction of the object's phase profile from one single interferogram. We investigate the theoretical background of "spiral interferometry" and suggest various demodulation techniques based on the processing of one single interferogram or multiple interferograms. PMID- 16715160 TI - Propagation of hollow Gaussian beams through apertured paraxial optical systems. AB - On the basis of the generalized Collins formula and the expansion of the hard aperture function into a finite sum of complex Gaussian functions, an approximate analytical formula for a hollow Gaussian beam propagating through an apertured paraxial stigmatic (ST) ABCD optical system is derived. Some numerical examples are given. Furthermore, by using a tensor method, we derive approximate analytical formulas for a hollow elliptical Gaussian beam propagating through an apertured paraxial general astigmatic ABCD optical system and an apertured paraxial misaligned ST ABCD optical system. Our results provide a convenient way for studying the propagation and transformation of a hollow Gaussian beam and a hollow elliptical Gaussian beam through an apertured general optical system. PMID- 16715161 TI - Numerical calculation of interparticle forces arising in association with holographic assembly. AB - Recent advances in dynamic holography have resulted in spatial light modulators capable of producing an almost limitless variety of field distributions from a single incident beam. Holographic assembly is a technique that exploits this capability to generate and control multiple foci that can be used to trap and manipulate nanoparticles. Although the forces associated with conventional optical tweezers are well understood, the effects arising from the more complicated interactions associated with holographic assembly are not. We present a general and flexible method, based on T matrix theory, for investigating these effects and use it to calculate the forces between particles in a variety of optical environments. PMID- 16715162 TI - Extension of the Williams-Clapper model to stacked nondiffusing colored coatings with different refractive indices. AB - We propose a model for predicting the reflectance and transmittance of multiple stacked nonscattering coloring layers that have different refractive indices. The model relies on the modeling of the reflectance and transmittance of a bounded coloring layer, i.e., a coloring layer and its two interfaces with neighboring media of different refractive indices. This model is then applied to deduce the reflectance of stacked nonscattering layers of different refractive indices superposed with a reflecting diffusing background that has its own refractive index. The classical Williams-Clapper model becomes a special case of the proposed stacked layer model. PMID- 16715163 TI - Diffraction of a Gaussian beam in a three-dimensional smoothly inhomogeneous medium: an eikonal-based complex geometrical-optics approach. AB - We present an ab initio account of the paraxial complex geometrical optics (CGO) in application to scalar Gaussian beam propagation and diffraction in a 3D smoothly inhomogeneous medium. The paraxial CGO deals with quadratic expansion of the complex eikonal and reduces the wave problem to the solution of ordinary differential equations of the Riccati type. This substantially simplifies the description of Gaussian beam diffraction as compared with full-wave or parabolic (quasi-optics) equations. For a Gaussian beam propagating in a homogeneous medium or along the symmetry axis in a lenslike medium, the CGO equations possess analytical solutions; otherwise, they can be readily solved numerically. As a nontrivial example we consider Gaussian beam propagation and diffraction along a helical ray in an axially symmetric waveguide medium. It is shown that the major axis of the beam's elliptical cross section grows unboundedly; it is oriented predominantly in the azimuthal (binormal) direction and does not obey the parallel-transport law. PMID- 16715164 TI - Time-resolved diffusing wave spectroscopy beyond 300 transport mean free paths. AB - We presented theoretical and experimental demonstrations of the possibilities of performing time-resolved diffusing wave spectroscopy: We successfully registered field fluctuations for selected photon path lengths that can surpass 300 transport mean free paths. Such performance opens new possibilities for biomedical optics applications. PMID- 16715165 TI - Fiber-based single-channel polarization-sensitive spectral interferometry. AB - We present a novel, to our knowledge, fiber-based single-channel polarization sensitive spectral interferometry system that provides depth-resolved measurement of polarization transformations of light reflected from a sample. Algebraic expressions for the Stokes parameters at the output of the interferometer are derived for light reflected from a birefringent sample by using the cross spectral density function. By insertion of a fiber-optic spectral polarimetry instrument into the detection path of a common-path spectral interferometer, the full set of Stokes parameters of light reflected from a sample can be obtained with a single optical frequency scan. The methodology requires neither polarization-control components nor prior knowledge of the polarization state of light incident on the sample. The fiber-based single-channel polarization sensitive spectral interferometer and analysis are demonstrated by measurement of phase retardation and fast-axis angle of a birefringent mica plate. PMID- 16715166 TI - Vectorial modal analysis of dielectric waveguides based on a coupled transverse mode integral equation. I. Mathematical formulation. AB - We propose a rigorous full-vector integral-equation formulation for analyzing modal characteristics of the complex, two-dimensional, rectangular-like dielectric waveguide that is divisible into vertical slices of one-dimensional layered structures. The entire electromagnetic mode field is completely determined by the y-component electric and magnetic field functions on the interfaces between slices. These interfacial functions are governed by a system of vector-coupled transverse-mode integral equations (VCTMIE) whose kernels are made of orthonormal sets of both TE-to-y and TM-to-y modes from each slice. To solve for the unknown functions, we construct sets of suitable expansion functions and turn VCTMIE into a nonlinear matrix equation via orthogonal projection. The eigenvectors of the matrix provide the mode field solutions of the complex dielectric waveguide. PMID- 16715167 TI - Vectorial modal analysis of dielectric waveguides based on a coupled transverse mode integral equation. II. Numerical analysis. AB - We present numerical implementation and verification of a rigorous full-vector, integral-equation formulation suitable for analyzing modal characteristics of complex, two-dimensional (2D) rectangular-like dielectric waveguides. By dividing the waveguide into vertical slices, a system of integral equations we call vector coupled transverse-mode integral equations (VCTMIE) is derived. The entire electromagnetic mode fields are completely determined by one-dimensional unknown field functions on the slice interfaces. To further reduce numerical computation, we expand these functions in terms of the guiding modes of a slab waveguide with a large normalized frequency. Through orthogonal projection the resulting nonlinear eigenvalue and eigenvector matrix formulation enables us to obtain the effective mode index with 10(-7) precision and to compute with high resolution the 2D vectorial mode field solutions of an open dielectric waveguide. We show stable and speedy convergence of our method as well as techniques to overcome the Gibbs phenomenon in the reconstruction of the transverse fields. PMID- 16715168 TI - Integral equation analysis of coupling in symmetric grating-assisted optical waveguides. AB - The propagation and coupling phenomena in grating-assisted optical couplers are analyzed by using an integral equation formulation and applying an entire-domain Galerkin technique. The proposed method constitutes a special type of the method of moments and provides high numerical stability and controllable accuracy. The electric field in the grating region is the unknown quantity and the resulting integral equation is subsequently solved by using Galerkin's method. The propagation constants of the guided waves are computed accurately by determining the singular points of the corresponding system's matrix. Numerical results regarding the propagation constants are presented for various coupler parameters, and the effect of the grating's physical and geometric characteristics on the coupling process is investigated. PMID- 16715169 TI - Mueller matrix description of polarization mode dispersion and polarization dependent loss. AB - We derive a differential equation that relates the Mueller matrices of an optical system at adjacent frequencies in the presence of polarization mode dispersion and polarization-dependent loss (PDL). We then demonstrate that a solution of this equation based on the Magnus expansion yields a description of the Mueller matrix in orders of the principal state vector that coincides with previously reported results for systems without PDL. PMID- 16715170 TI - Polarization mode dispersion in short fiber lengths. AB - We perform Monte Carlo simulations of the statistical properties of the differential group delay for fiber lengths less than and of the order of the birefringence correlation length. We find that the manner in which quantities related to the polarization mode dispersion evolve along the fiber depends significantly on the model assumed for the form of the birefringence. PMID- 16715171 TI - Non-Hermitian polarizers: a biorthogonal analysis. AB - The non-Hermitian operators of the ideal nonorthogonal multilayer optical polarizers are spectrally analyzed in the framework of skew-angular biorthonormal vector bases. It is shown that these polarizers correspond to skew projectors and their operators are generated by skew projectors, exactly as the canonical ideal polarizers correspond to Hermitian projectors. Thus the common feature of all the polarizers (Hermitian and non-Hermitian) is that their "nuclei" are (orthogonal or skew) projectors--the generating projectors. It is shown that if these nonorthogonal polarizers are looked upon as variable devices, two kinds of degeneracy may occur for suitable values of the inner parameter of the device: The corresponding operators may become normal (more precisely, Hermitian) or, on the contrary, very pathological--defective and singular. In the first case their eigenvectors and biorthogonal conjugate eigenvectors collapse into a unique pair of eigenvectors; in the second case their eigenvectors (as well as their biorthogonal conjugates) collapse into a single vector. PMID- 16715172 TI - Eigenmode expansions using biorthogonal functions: complex-valued Hermite Gaussians: comment. AB - In this communication, certain comments are made on the paper by Kostenbauder et al., "Eigenmode expansions using biorthogonal functions: complex-valued Hermite Gaussians,"J. Opt. Soc. Am. A 14, 1780 (1997). PMID- 16715174 TI - Spectral and polarization characteristics of the nonspherically decaying radiation generated by polarization currents with superluminally rotating distribution patterns: comment. AB - I repeat my (very short and easy) disproof of the recurring main claim of Ardavan et al. [J. Opt. Soc. Am. A 21, 858 (2004)] that a smooth source of electromagnetic fields moving in a confined region can generate an intensity decaying more slowly than the inverse square of the distance away ("nonspherical" decay). The field is not isotropic, so energy conservation is not enough to dismiss the claim. Instead my disproof follows directly from Maxwell's equations, supplying an upper bound with inverse square decay on the intensity. It therefore applies under all circumstances, quite irrespective of any fast or slow motion of the source. Despite the falsity of the main claim, the derivation of the uniform approximation to the Green function for superluminal circulation, which was needed for the claim and is based on the previous work of the first author, is valid. Its validity, importantly, extends significantly beyond the regime envisaged by the authors, and it stands as a basic result of superluminal circulation. PMID- 16715176 TI - Origination of an X-linked testes chimeric gene by illegitimate recombination in Drosophila. AB - The formation of chimeric gene structures provides important routes by which novel proteins and functions are introduced into genomes. Signatures of these events have been identified in organisms from wide phylogenic distributions. However, the ability to characterize the early phases of these evolutionary processes has been difficult due to the ancient age of the genes or to the limitations of strictly computational approaches. While examples involving retrotransposition exist, our understanding of chimeric genes originating via illegitimate recombination is limited to speculations based on ancient genes or transfection experiments. Here we report a case of a young chimeric gene that has originated by illegitimate recombination in Drosophila. This gene was created within the last 2-3 million years, prior to the speciation of Drosophila simulans, Drosophila sechellia, and Drosophila mauritiana. The duplication, which involved the Ballchen gene on Chromosome 3R, was partial, removing substantial 3' coding sequence. Subsequent to the duplication onto the X chromosome, intergenic sequence was recruited into the protein-coding region creating a chimeric peptide with approximately 33 new amino acid residues. In addition, a novel intron containing 5' UTR and novel 3' UTR evolved. We further found that this new X linked gene has evolved testes-specific expression. Following speciation of the D. simulans complex, this novel gene evolved lineage-specifically with evidence for positive selection acting along the D. simulans branch. PMID- 16715178 TI - Dynamic analysis of DNA damage by flow cytometry and FISH. AB - The micronucleus assay, developed to assess DNA damage induced by noxious agents, supplies information on whether the damage is due to clastogenic or aneugenic action. Although it is the test that can be used to assess agents' toxicity, it cannot provide information on the molecular events that result in the induction of micronuclei. To study the molecular events, the combination of both microscopic and analytical techniques is required. Flow-sorting induced micronuclei, based on their DNA content, in combination with chromosomal FISH and other molecular techniques, may provide information on these events. PMID- 16715177 TI - Factors influencing engagement into interventions for adaptation to HIV in African American women. AB - Factors that may influence engagement into a family-ecological psychosocial intervention and a nondirective psychosocial intervention designed for HIV+ asymptomatic women were examined. Participants were 136 HIV+ African American women. Participant characteristics and therapeutic alliance were examined as possible predictors of engagement. Both participant characteristics and therapeutic alliance had some power in predicting engagement. However, fewer participant characteristics than expected were statistically significant. Statistically significant results indicate that women who had more daily hassles, more distress, more social support, and more disagreements with their spouse were more likely to engage in the intervention. The strongest predictor of engagement was therapeutic alliance, indicating the importance of the alliance between the HIV+ participant and the interventionist. The importance of these findings is discussed. PMID- 16715179 TI - ECG changes in 8-year-old boy with pulmonary edema after head injury. AB - This is a case story of an 8-year-old boy with no prior history of cardiac disease who developed acute pulmonary edema with ECG changes similar to transmural myocardial infarction after basilar skull fracture. Biochemical evaluation showed elevated total creatine kinase activity -1,350 U/L with 12% MB isoenzyme fraction. The brain scan on admission showed cerebral edema with ethmoidal sinuses hemorrhage. Neurogenic pulmonary edema following CNS damage is an extremely rare entity in the pediatric population and there are few reports. There are many proposed mechanisms and explanations of its origin. Based on previous reports and experimental studies, the cause of "neurogenic" pulmonary edema may be of cardiac as well as of noncardiac origin. PMID- 16715180 TI - Serial correlation in lateralized choices of hand and target. AB - We investigated how lateralized choices of hand and target are influenced by previous behavior. Three monkeys retrieved food pellets following cues indicating the location of available food pellet targets, and the hand that could be used to acquire a target. In pseudo-randomized trials, the monkeys could retrieve food pellet targets only on their right side, only on their left side, or their choice of either right or left side, using only their right hand, only their left hand, or their choice of either hand. We examined separately the patterns of serial correlation in target choices and hand choices. Although individual monkeys showed overall laterality preferences, instead of repeatedly using the preferred hand, we found that the monkeys tended to switch hands in successive trials. This serial correlation in hand choice was stronger and more robust than serial correlation in target choice. Furthermore, the pattern of serial correlation for target choice closely resembled that of serial correlation for hand choice when the animal was allowed to choose both target and hand, but only when the target cue was presented before the hand cue. These results suggest that when cued to choose a hand first, the monkeys tended to make a separate decision as to whether to switch their target choices or not, whereas their decisions to switch hands and targets were linked more tightly if the animal was cued to choose a target first. PMID- 16715181 TI - The future of epidemiology: methodological challenges and multilevel inference. AB - A decade ago there was considerable debate about the appropriate objectives and paradigms of modern epidemiologic research. One concern put forth in these debates was that "risk factor epidemiology" might be forcing our field to focus more on individuals and less on populations and public health. Today, most epidemiologists acknowledge that public health is influenced by both population level and individual-level determinants. Ecologic studies are valuable tools for generating hypotheses and addressing group-level determinants of disease risk. Traditional risk factor studies and genomic studies have helped establish the multifactorial concept of disease causation. Individual-level studies also have provided the biomedical community with hypotheses that have stimulated research into disease mechanisms that have led to reductions in morbidity and mortality for diseases such as HIV/AIDS, cardiovascular disease, and cancer. Current debates about the role of genomic data in epidemiology and public health mirror the debates about risk factor epidemiology one decade ago. Genomic variation is measured at the individual level, but how this variation is maintained in human populations is a group-level (population) phenomenon that is worthy of epidemiologic investigation in its own right. Multilevel epidemiology seeks to understand multiple levels of inference, from genes to individuals to populations and could combine hypothesis-driven research with aspects of data mining. Multilevel epidemiology calls for the study of health and disease determinants defined at the population level and individual level for a more comprehensive strategy to understanding human disease etiology. With the continued development of multilevel statistical methods and the advent of data mining, the technical constraints of the past will become less relevant to the next generation of epidemiologists who wish to embrace a more multilevel epidemiology. PMID- 16715182 TI - [The role of health care reporting for the planning of long-term care at the community level]. AB - A key element in the demographic transition process is the increase in the number of very old people (80+) leading to a rising need for long-term care. For the municipalities the efficient organisation of the local support for senior citizens is an important task for legal, political and financial reasons. The local planning process must be based on systematic reporting about long-term care in the community. This report must contain fundamental facts about the demographic situation, the health care system, including the quality of care in nursing homes, by ambulatory services and families as well as the resulting costs in the local welfare budget. Comparing the problem to the methods in local health promotion it is favourable to establish an office to manage the planning process and a committee for matters of care. Committee members should be all relevant stakeholders of the local health, the care and the social sector. The first priority is to achieve the participation of patients, their relatives and the local politicians to agree on targets and measures in the planning process. Key targets are the prevention of risks for long-term care, to secure the quality of care and the preference for ambulatory services, optimisation of local cooperation and minimising the costs for the community. The whole process should be guided by these targets for the provision of care. PMID- 16715183 TI - Comparative study on in vitro effects of homocysteine thiolactone and homocysteine on HUVEC cells: evidence for a stronger proapoptotic and proinflammative homocysteine thiolactone. AB - Hyperhomocysteinemia is an independent risk factor for the development of atherosclerosis. However the underlying mechanisms responsible for endothelial cell injury with increased plasma concentration of homocysteine or homocysteine derivatives remains still incompletely elucidated. In this study, we investigated the ability of homocysteine (Hcy) and homocysteine thiolactone (HcyT) to induce cell death and IL-8 secretion in primary human umbilical vein endothelial cells (HUVEC). Hcy and HcyT were both cytotoxic and capable of promoting cell death, as measured by caspase-3 activation and DNA fragmentation. ELISA assays clearly demonstrated that Hcy and HcyT strongly activated IL-8 release. Furthermore, our results showed that HcyT was much more efficient than Hcy in activating caspase-3 or in inducing IL-8 secretion. The use of antioxidants such as vitamin C and vitamin E strongly but not completely reduced programmed cell death and chemokine release suggesting that other pathways different than reactive oxygen species are also involved. This study suggests that Homocysteine derivatives like HcyT might possess stronger cytotoxicity and pro-inflammatory properties and that Hcy derivatives levels should therefore be more taken into account during diagnostics. PMID- 16715184 TI - Influence of nicorandil on the pharmacodynamics and pharmacokinetics of gliclazide in rats and rabbits. AB - Chronic diabetes precipitates ischaemic heart disease (IHD) and many other disorders. IHD inturn is shown in the form of angina initially. According to EUROPA study, the incidence of angina is high in type II diabetics. Gliclazide, a second generation sulphonylurea derivative is widely used in the treatment of type-II diabetes and is known to release insulin by K(+) channel inhibition. Nicorandil, a newer antianginal drug widely used now a days acts by opening potassium channels in the cardiac muscle cell and also by releasing nitric oxide. However its action on pancreatic cell K(+) channel is not known. Since there is possibility for drug interaction leading to decreased activity of gliclazide the present study was conducted to evaluate the effect of the combination. Studies in normal and alloxan induced diabetic rats were conducted with oral doses of 2 mg/kg bd. wt. of gliclazide, 1.8 mg/kg bd. wt. of nicorandil and their combination with adequate washout periods in between treatments. Studies in normal rabbits were conducted with 5.6 mg/1.5 kg bd. wt. of gliclazide, 1.4 mg/1.5 kg bd. wt. of nicorandil and their combination given orally. Blood samples were collected in rats from retro orbital puncture at 0, 1, 2, 3, 4, 6, 8, 10 and 12 h and by marginal ear vein puncture in rabbits at 0, 1, 2, 3, 4, 6, 8, 12, 16, 20 and 24 h. All the blood samples were analysed for glucose by GOD/POD method. The blood samples of rabbits were analysed by HPLC for gliclazide. Gliclazide produced hypoglycaemic/antidiabetic activity in normal and diabetic rats with peak activity at 1 h and 8 h and hypoglycaemic activity in normal rabbits at 3 h, while nicorandil alone produced significant hyperglycaemia at 4 h and reduced the effect of gliclazide with no significant change in pharmacokinetics when administered in combination. The interaction observed appears to be pharmacodynamic at the receptor level as expected. PMID- 16715185 TI - Bacterial community along a historic lake sediment core of Ardley Island, west Antarctica. AB - The bacterial community in a historic lake sediment core of Ardley Island, Antarctica, spanning approximately 1,600 years, was investigated by molecular approaches targeting the 16S rRNA gene fragments. The cell number in each 1 cm layer of the sediment core was deduced through semi-quantification of the 16S rRNA gene copies by quantitative competitive PCR (QC-PCR). It was found that the total bacterial numbers remained relatively stable along the entire 59 cm sediment core. Denaturing Gradient Gel Electrophoresis (DGGE) analysis and sequencing of PCR-amplified 16S rRNA gene fragments were performed to analyze the bacterial diversity over the entire column. Principle coordinates analysis suggested that the bacterial communities along the sediment core could be separated into three groups. There were obvious bacterial community shift among groups of 1-20 cm, 21-46 cm and 46-59 cm. Diversity indices indicated that the bacterial community in the 21-46 cm depth showed the highest species diversity and uniformity. The main bacterial groups in the sediments fell into 4 major lineages of the gram-negative bacteria: the alpha, gamma and delta subdivision of Proteobacteria, the Cytophaga-Flavobacteria-Bacteroides, and some unknown sequences. The gram-positive bacteria Gemmatimonadetes, Firmicutes and Actinobacteria were also detected. The results demonstrated the presence of highly diverse bacterial community population in the Antarctic lake sediment core. And the possible influence of climate and penguin population change on the bacterial community shift along the sediment core was discussed. PMID- 16715186 TI - B-type natriuretic peptide levels in patients with paroxysmal lone atrial fibrillation. AB - To investigate the levels of B-type natriuretic peptide (BNP) in patients with atrial fibrillation (AF) but without structural heart disease, we measured plasma BNP concentration in 61 consecutive AF patients and in 61 age- and sex-matched healthy subjects. Plasma BNP concentration in the AF group was significantly higher than in the control group (121+/-32 vs 41+/-12 pg/ml, P<0.001). Logistic regression analysis showed that age (r=0.66, P<0.001), left atrial diameter (r=0.59, P<0.01), and a history of AF (r=0.72, P<0.001) were independent predictors of elevated BNP. We concluded that BNP was elevated in patients with paroxysmal lone AF. The clinical significance of BNP elevation in these patients requires further investigation. PMID- 16715187 TI - Safety of gadodiamide mixed with a small quantity of iohexol in patients with impaired renal function undergoing coronary angiography. AB - Gadolinium-based contrast agents can be used in diagnostic and interventional angiography, and are safe in recommended doses in patients with impaired renal function, but the image quality is usually unsatisfactory. The objective of the present study is to evaluate the safety of gadolinium mixed with a small quantity of iodine-based contrast agent and the image quality of this mixture in patients with renal insufficiency undergoing diagnostic coronary angiography. Forty-two patients with baseline creatinine level>1.5 mg/dl were randomized into gadolinium or iodine groups. In the gadolinium group gadodiamide was mixed with 1:3 iohexole and in the iodine group only this agent was used as contrast material. Peak creatinine levels 48-72 h after the procedure were measured. The primary end point was the development of contrast-induced nephropathy (CIN), defined as >25% increase of baseline creatinine levels, and the secondary end points were absolute and relative changes in baseline creatinine levels. Total contrast volumes used were not different between groups (57+/-11 ml in gadolinium and 55+/ 10 ml in iodine; P=0.68). Mean creatinine level did not increase significantly in the gadolinium group (from 1.9+/-0.3 to 1.9+/-0.4 mg/dl; P=0.06), but did in the iodine group (from 2.0+/-0.4 to 2.3+/-0.5 mg/dl; P=0.001). No patient had CIN in the gadolinium group whereas 5 (23%) patients had this phenomenon in the iodine group (P=0.048). Contrast regimen was very well tolerated in the gadolinium group, with only transient headache in two patients. Gadodiamide mixed with a small quantity of iohexol is safe in patients with azotemia undergoing diagnostic coronary angiography. The image qualities obtained with this combination are also satisfactory in all of the cases. Further evaluation of the safety of this technique is warranted, especially in other types of diagnostic and interventional procedures in which a higher amount of contrast dye is needed. PMID- 16715188 TI - Preprocedural statin therapy reduces the risk and extent of cardiac biomarker release following percutaneous coronary intervention. AB - This study evaluates the association between statin therapy in patients treated by percutaneous coronary intervention (PCI) for stable angina pectoris and postinterventional myocardial injury with subsequent long-term clinical outcome. Prospectively collected data on 400 consecutive patients with stable angina pectoris or evidence of inducible myocardial ischemia were analyzed. The incidence of myocardial infarction based on postinterventional release of troponin I>1.5 ng/ml was 12% in the statin pretreated patients and 20% in those not pretreated with statin therapy (P=0.04, odds ratio 1.84, 95% confidence interval 1.06-3.21). Of the patients experiencing a post-PCI troponin elevation>1.5 ng/ml, those pretreated with a statin pre-PCI had a lesser troponin elevation compared with those not receiving a statin pre-PCI (median: 2.9 ng/ml [1.9-11.5] vs 5.0 ng/ml [3.1-8.8]; P<0.001). In the multivariate model, preprocedural statin therapy was identified as the only independent negative predictor of procedure-related myocardial necrosis based on postprocedural troponin elevation. In the 21-month follow-up period, statin pretreated patients were observed to have fewer deaths, revascularizations, or myocardial infarction; however, this difference was not statistically significant. These results suggest that pretreatment with statins in patients undergoing PCI for stable angina pectoris reduces the risk and extent of procedure-related myocardial injury measured by troponin release. PMID- 16715189 TI - The acute cardioprotective effect of glucocorticoid in myocardial ischemia reperfusion injury occurring during cardiopulmonary bypass. AB - The purpose of this study was to evaluate the acute cardioprotective effect of high-dose methylprednisolone (25 mg/kg) in the controlled in vivo model of myocardial ischemia-reperfusion injury occurring during cardiopulmonary bypass. Forty nondiabetic male patients with three-vessel disease undergoing first-time bypass surgery were enrolled for this double-blind prospective study. Patients were randomized to be given 25 mg/kg methylprednisolone (Group I) and saline (Group II) 1 h before cardiopulmonary bypass. The levels of cardiac troponin-I (cTnI) were used as a marker of myocardial tissue damage in myocardial ischemia reperfusion injury. The cTnI levels were measured before surgery, at the second hour after cardiopulmonary bypass, at the 6th and 24th hours, and 5th day postoperatively. There was no significant difference between the two groups in respect to the duration of ischemia and reperfusion. The preoperative cTnI levels were 0.22+/-0.29 ng/ml in Group I and 0.23+/-0.28 ng/ml in Group II. cTnI levels increased to 2.40+/-1.0 ng/ml in Group I and 3.19+/-0.88 ng/ml in Group II at the 2nd hour after cardiopulmonary bypass. When the differences between T1 and T0 level that showed the amount of troponin release occurring due to ischemia repefusion injury was calculated and then compared, there was a significant difference between Groups I and II (P=0.024). The cTnI levels measured at 6 h after CPB were 1.98+/-0.63 ng/ml in Group I and 2.75+/-1.15 ng/ml in Group II (P=0.049). cTnI levels decreased to 0.22+/-0.10 ng/ml in Group I and 0.49+/-0.25 ng/ml in Group II on the postoperative day 5 (P=0.0001). Univalent regression analysis showed that preoperative high-dose corticosteroid usage decreased the troponin release in about 12% and this effect was statistically significant (R2=0.12, P<0.05). A single dose of intravenous methylpredisolone (25 mg/kg) given 1 h before ischemia reduced myocardial ischemia-reperfusion injury. These results demonstrated that the acute cardioprotective effect of corticosteroids has much potential in the future for reducing ischemia-reperfusion injury occurring during cardiopulmonary bypass when it is inevitable. PMID- 16715190 TI - Is the coronary flow velocity reserve improvement after aortic valve replacement for aortic stenosis transient? Results of a 3-year follow-up. AB - The coronary flow velocity reserve (CFR) is decreased in patients with aortic valve stenosis (AS). The aim of the present prospective study was to examine the CFR in AS patients with normal epicardial coronary arteries during a 3-year follow-up of aortic valve replacement (AVR). A total of 30 AS patients (17 women and 13 men) were enrolled in this prospective follow-up study; they all had a normal coronary angiogram and underwent dipyridamole stress transesophageal echocardiography (STEE) for CFR measurement before AVR. Nine of them participated in this 3-year follow-up study, and all patients were examined 16 months and 3 years after AVR by STEE. The CFR of AS patients was decreased before AVR. Different extents of diastolic coronary flow velocity reduction (resting and posthyperaemic) were observed resulting in a significant CFR improvement, despite the low number of patients during the 16-month follow-up. A further posthyperaemic flow velocity reduction was demonstrated after the 3-year follow up, resulting in a significant CFR impairment. The CFR decreased in 5 cases, but remained practically unchanged in 3 after the first CFR measurement; only the CFR of 1 patient improved minimally. The main finding of the current study is that the CFR improvement 16 months after AVR, which paralleled the regression of the left ventricular hypertrophy, was found to be transient. Coronary flow velocity reserve impairment was demonstrated after the long-term (3-year) follow-up in most of the patients, which could not be explained by extravascular compressive forces. PMID- 16715191 TI - Effects of verapamil and nifedipine on different parameters in lipopolysaccharide induced septic shock. AB - Septic shock has a high mortality rate due to the hypotension and circulatory disorder that occurs during its pathogenesis. Recently, humoral factors such as cytokines and nitric oxide became important in the complex pathophysiology of septic shock because there is a close relationship between the determined levels of these humoral factors and the responses to the therapy and survival periods. Verapamil and nifedipine are calcium channel blockers commonly used in the pharmacotherapy of cardiovascular disorders. In the present study these drugs were investigated in the rat septic shock model. In vivo hemodynamic parameters were recorded using a data acquisition system in endotoxin-induced septic shock in rats. The animals were followed for 5 h and blood pressure, rectal temperature, and ECG were recorded. Blood samples were collected at 1 h and 5 h time points after the injection of endotoxin, and serological samples were stored at -25 degrees C. Subsequently, tumor necrosis factor-alpha, interleukin-10 (enzyme-linked immunosorbent assay), and nitrite (Griess reagent) were determined in these serological samples. Significant correlations were observed between these humoral factors and the disordered hemodynamic factors. A reversal of changes was observed in the levels of serum cytokines, nitrite levels, and hemodynamic parameters with verapamil and nifedipine preadministration (P<0.05). Additionally, superoxide dismutase (SOD), catalase, and malondialdehyde (MDA) were determined in livers obtained from these animals at the end of the experiments, and these results were compared to hemodynamic parameters and cytokines. Nifedipine and verapamil increased the levels of MDA and SOD but did not change catalase activity. PMID- 16715192 TI - Inhalation of nebulized nitroglycerin, a nitric oxide donor, for the treatment of pulmonary hypertension induced by high pulmonary blood flow. AB - This study explores the effect and mechanisms of inhalation of nebulized nitroglycerin (Neb-NTG) on pulmonary hypertension induced by high pulmonary blood flow. An aortocaval shunt was produced in rats. Twelve weeks after the operation, rats started to inhale Neb-NTG. After 15 weeks, pulmonary and systemic hemodynamics as well as pathological changes were measured in all animals. Inhalation of Neb-NTG was able not only to markedly attenuate pulmonary artery pressure without impacting systolic pressure but also to ameliorate muscularization of small pulmonary arteries. The relaxation response of pulmonary artery ring to acetylcholine in shunt rats and Neb-NTG-treated rats was decreased. However, Neb-NTG did not impact the relaxation response of pulmonary artery ring to both nitroprusside and nitroglycerin (NTG). Neb-NTG successfully inhibited the increased expression of pulmonary artery smooth muscle cell (PASMC) proliferative cell nuclear antigen, collagen I, and collagen III, as well as pulmonary artery urotensin-II in shunt rats with high pulmonary blood flow. Neb NTG selectively ameliorated pulmonary hypertension as well as pulmonary vascular structural remodeling induced by high pulmonary blood flow. PMID- 16715193 TI - Autologous bone marrow cell transplantation improves left ventricular function in rabbit hearts with cardiomyopathy via myocardial regeneration-unrelated mechanisms. AB - Recent studies suggest transplanted bone marrow cells (BMCs) can be used to reconstitute coronary vessels and myocardium following acute myocardial infarction, thereby improving cardiac function. We sought to investigate the therapeutic potential of BMC transplantation in the treatment of nonischemic cardiomyopathy. Experimental cardiomyopathy was produced by treating rabbits for 8 weeks with doxorubicin (2 mg/kg per week). Two weeks after the treatment was finished, freshly aspirated BMCs or an equivalent volume of phosphate-buffered saline was injected directly into the left ventricular free wall. Four weeks later, heart function was examined during perfusion on a Langendorff apparatus. Left ventricular developed pressure and +/-dp/dt were significantly better in the transplantation group, among which echocardiography also showed significantly better ejection fractions. In addition, left ventricular weights as a fraction of body weight and left ventricular wall thicknesses were both lower in rabbits transplanted with BMCs than in controls. Immunohistochemical analyses carried out 2 weeks after transplantation showed no new myocardium and a very small number of endothelial cells originating from BMCs. On the other hand, immunoblotting revealed upregulated expression of transforming growth factor-beta1 and downregulated expression of matrix metalloproteinase-1 and tumor necrosis factor alpha following BMC transplantation. In conclusion, autologous BMC transplantation into cardiomyopathic rabbit hearts ameliorates the decline in ventricular function without regenerating cardiomyocytes, most likely by altering expression of various cytokines. PMID- 16715194 TI - A permanent junctional reciprocating tachycardia with an atypically located accessory pathway successfully ablated from within the middle cardiac vein. AB - Permanent form of junctional reciprocating tachycardia (PJRT) is an uncommon form of atrioventricular re-entrant tachycardia due to an accessory pathway characterized by slow and decremental retrograde conduction. The majority of accessory pathways in PJRT are localized in the posteroseptal zone. Despite the high success rate, failure may occur during endocardial radiofrequency catheter ablation due to epicardial insertion of the accessory pathway. We report a case of PJRT in a 25-year-old man in whom the accessory pathway was located epicardially in the posteroinferior region and ablated from within the middle cardiac vein by radiofrequency catheter ablation. PMID- 16715195 TI - Left ventricular aneurysm associated with isolated noncompaction of the ventricular myocardium. AB - A 66-year-old woman was admitted to our hospital because of left ventricular failure and nonsustained ventricular tachycardia. Two-dimensional echocardiography demonstrated prominent trabeculations and deep intertrabecular recesses, findings consistent with noncompaction of the ventricular myocardium. Myocardial perfusion scintigraphy demonstrated a defect in the anterobasal left ventricular segment. Coronary angiogram was normal, but the left ventriculogram showed an aneurysm in the anterior myocardial segments. This is the first reported case with isolated noncompaction of the ventricular myocardium associated with left ventricular aneurysm. PMID- 16715196 TI - Noncompaction of the ventricular myocardium combined with polycystic kidney disease. AB - Noncompaction of the ventricular myocardium (NVM) is a rare cardiac abnormality of unknown etiology. The condition is characterized by prominent and excessive trabeculations in a ventricular wall segment, with deep intertrabecular spaces perfused from the ventricular cavity. Polycystic kidney diseases are characterized by the formation of multiple cysts in the kidneys and liver and, less frequently, in the pancreas. Cardiovascular abnormalities including hypertension, mitral valve prolapse, and intracranial aneurysms are also frequently recognized. However, polycystic kidney disease and isolated ventricular noncompaction have not previously been correlated. Here, we describe one case of isolated noncompaction of ventricular myocardium with polycystic kidney disease, coupled with a progressive worsening of heart failure. We confirmed these abnormalities using contrast echocardiography, abdominopelvic computed tomography, and cardiac magnetic resonance imaging. PMID- 16715197 TI - Acrobatic vascular disease. AB - Athletes performing repetitive movements can develop arterial injuries. These are uncommon. We present a case of bilateral subclavian/axillary artery stenosis in a female patient referred to the rapid access chest pain clinic with a presumptive diagnosis of angina. Her symptoms were related to limb ischemia rather than cardiac insufficiency. Detailed history from this patient revealed that she had spent 8 years working as an acrobat being swung by the arms twice daily during performances. PMID- 16715198 TI - Myocardial and skeletal muscle biopsy in left ventricular hypertrabeculation/noncompaction. PMID- 16715199 TI - [Diagnostic imaging for connective tissue diseases]. AB - Guidelines and diagnostic algorithms for the introduction of diagnostic procedures for connective tissue diseases have not yet been established and standardized. Diagnosis is often based on the patients's typical history, clinical symptoms and specific laboratory profiles. The use of diagnostic imaging procedures is relevant for differential diagnosis, for the assessment of disease activity and organ involvement, for defining the prognosis and monitoring responses to therapy and side effects. Experience with these techniques and diagnostic procedures, their application, indications and analysis represent the platform for rational and optimal diagnosis as well of defined therapy stratification. PMID- 16715201 TI - A new POLG1 mutation with peo and severe axonal and demyelinating sensory-motor neuropathy. AB - BACKGROUND: Progressive external ophthalmoplegia (PEO) is a mitochondrial disorder associated with defective enzymatic activities of oxidative phosphorylation (OXPHOS), depletion of mitochondrial DNA (mtDNA) and/or accumulation of mtDNA mutations and deletions. Recent positional cloning studies have linked the disease to four different chromosomal loci. Mutations in POLG1 are a frequent cause of this disorder. METHODS: We describe two first-cousins: the propositus presented with PEO,mitochondrial myopathy and neuropathy, whereas his cousin showed a Charcot- Marie-Tooth phenotype. Neurophysiological studies, peroneal muscle and sural nerve biopsies, and molecular studies of mtDNA maintenance genes (ANT1, Twinkle, POLG1, TP) and non dominant CMT-related genes (GDAP1, LMNA, GJB1) were performed. RESULTS: A severe axonal degeneration was found in both patients whereas hypomyelination was observed only in the patient with PEO whose muscle biopsy specimen also showed defective OXPHOS and multiple mtDNA deletions. While no pathogenetic mutations in GDAP1, LMNA, and GJB1 were found, we identified a novel homozygous POLG1 mutation (G763R) in the PEO patient. The mutation was heterozygous in his healthy relatives and in his affected cousin. CONCLUSIONS: A homozygous POLG1 mutation might explain PEO with mitochondrial abnormalities in skeletal muscle in our propositus, and it might have aggravated his axonal and hypomyelinating sensory-motor neuropathy. Most likely, his cousin had an axonal polyneuropathy with CMT phenotype of still unknown etiology. PMID- 16715202 TI - Opsoclonus-myoclonus syndrome in an adult with malignant melanoma. PMID- 16715200 TI - Infection-associated encephalopathies: their investigation, diagnosis, and treatment. AB - Reduced level of consciousness is a common clinical finding in acutely sick patients. In the majority of cases a cause for the encephalopathy is readily identifiable,whilst in a minority the aetiology is more difficult to ascertain. Frequently the onset of encephalopathy is associated with, or follows, infection. The mechanisms through which infection leads to encephalopathy are diverse. They range from direct microbial invasion of the brain or its supporting structures, to remote, infection-triggered mechanisms such as acute disseminated encephalomyelitis. Most common however, is the encephalopathy caused through a remote effect of systemic sepsis-septic encephalopathy. This article discusses the clinical presentation and underlying pathogeneses of the acute encephalopathies associated with infection, aiming to aid both their recognition and treatment. PMID- 16715204 TI - Comparison of communities of stored product mites in grain mass and grain residues in the Czech Republic. AB - In storage facilities one can find grain either in stored grain mass or in grain residues in the store corners or machinery. Although it is claimed that grain residues are serious pest reservoirs since they harbor numbers of stored product arthropods and are connected via continuous emigration with grain mass, the documentation for this is not convincing. Therefore in 78 selected grain stores, we simultaneously sampled the grain mass and residues in order to compare concurrent mite communities in these two different habitats. We found 30 species in about 614,000 individuals in residues and 23 species in about 20 000 individuals in grain mass. Canonical correspondence analysis (CCA) of transformed abundance data showed differences in the communities of mites in grain mass and residues: (i) species associated to grain residues (e.g. Tyrophagus longior, Tydeus interruptus, Acarus farris and Cheyletus eruditus) and (ii) species associated to both grain mass and grain residues (e.g. Tarsonemus granarius, Acarus siro, Tyrophagus putrescentiae, Lepidoglyphus destructor and Cheyletus malaccensis). Although the residue samples had more mites and higher species diversity than the stored grain mass, no correlation in mite abundance and species numbers between samples from grain residues and grain mass was found, thereby indicating low connectivity of these two habitats. PMID- 16715205 TI - Striatal FP-CIT uptake differs in the subtypes of early Parkinson's disease. AB - In idiopathic Parkinson's disease (PD), a tremor-dominant type (TDT), an akinetic rigid type (ART), and a mixed type (MT) are distinguished. We compared cerebral [I-123]FP-CIT SPECT in the PD subtypes (67 patients Hoehn and Yahr stage 1:26 with ART, 19 with MT, 22 with TDT). We measured the ratios putamen/occipital lobe binding and caudate nucleus/occipital lobe binding. Parkinsonian motor symptoms were quantified by UPDRS motor scale. In both putamen and caudate nucleus contralateral to the clinically affected body side TDT patients showed a significantly higher FP-CIT uptake than ART or MT patients (ANOVA; p<0.01). Contralateral putamen and caudate nucleus FP-CIT uptake correlated significantly with severity of rigidity (p<0.01) and hypokinesia (p<0.01) but not with severity of resting or postural tremor (p>0.05). The missing correlation between striatal FP-CIT uptake and tremor suggests, that further systems besides the nigrostriatal dopaminergic system may contribute to generation of parkinsonian tremor. PMID- 16715206 TI - Contralateral prefrontal projections in gerbils mature abnormally after early methamphetamine trauma and isolated rearing. AB - As previously shown, a miswiring of ipsilateral prefrontal projections after methamphetamine (MA) intoxication and/or isolated rearing (IR) may serve as a model of so-called "dysconnection" in human schizophrenia. We here find that deep prefrontal projections to contralateral targets were drastically reduced by both MA and IR alone, but remained equally dense if both impairments cumulated. Projections from superficial layers were not altered by MA and/or IR. These findings confirm that the normal intercortical integration of information is compromised in this animal model of schizophrenia. PMID- 16715207 TI - Proton magnetic resonance spectroscopy in developmentally delayed young boys with or without autism. AB - OBJECTIVE: The aim of the present study is to investigate whether brain metabolism of boys with autism spectrum disorder (ASD) is altered compared to boys with a developmental delay without autism if corrected for patient age and developmental level. STUDY DESIGN: 25 boys with ASD (with or without concurrent mental retardation) and 12 boys without ASD with mental retardation or language disorder underwent proton magnetic resonance spectroscopy. All analyses were performed with chronological age and developmental level as independent variables. RESULTS: No metabolic differences were found between boys with ASD and without ASD. CONCLUSIONS: Our findings do not replicate previous reports of differences in NAA, Cho and Cr levels in ASD. PMID- 16715208 TI - Staurosporin induces neurite outgrowth through ROS generation in HN33 hippocampal cell lines. AB - Staurosporin, a specific inhibitor of PKC, is widely used in studies of signal transduction pathways. Previous studies have shown that staurosporin induces neurite outgrowth, but the underlying mechanisms remain unclear. Here we report that staurosporin induces neurite outgrowth in HN33 hippocampal cells. Two other PKC inhibitors, Go 6976 (specific for alpha- and beta-isoforms) and rotterlin (a selective inhibitor of PKC delta), have no neuritogenic effect. In addition, staurosporin specifically increases ROS generation. NAC, which inhibits the generation of ROS, suppresses the staurosporin-induced neurite outgrowth in HN33 cells. Further, H(2)O(2) causes neurite outgrowth. Taken together, these results confirm a neuritogenic effect of staurosporin and point to ROS as the signal mediator of staurosporin-induced neurite outgrowth in HN33 hippocampal cells. Theme: Development and regeneration Topic: Neurotrophic factors: receptors and cellular mechanisms. PMID- 16715209 TI - 7-Ketocholesterol enhances 1-methyl-4-phenylpyridinium-induced mitochondrial dysfunction and cell death in PC12 cells. AB - The present study investigated the promoting effect of oxysterol 7 ketocholesterol against the cytotoxicity of 1-methyl-4-phenylpyridinium (MPP(+)) in differentiated PC12 cells. 7-Ketocholesterol significantly enhanced the MPP(+) induced nuclear damage, decrease in the mitochondrial transmembrane potential, cytosolic accumulation of cytochrome c, activation of caspase-3, increase in the formation of reactive oxygen species and depletion of GSH. N-Acetylcysteine, ascorbate, trolox, carboxy-PTIO and Mn-TBAP reduced the cytotoxic effect of MPP(+) in the presence of 7-ketocholesterol. The results indicate that 7 ketocholesterol shows a synergistic effect against the cytotoxic effect of MPP(+). 7-Ketocholesterol may enhance the MPP(+)-induced viability loss in PC12 cells by promoting the mitochondrial membrane permeability change, release of cytochrome c and subsequent activation of caspase-3, which is associated with the increased formation of reactive oxygen species and depletion of GSH. The findings suggest that 7-ketocholesterol as a promoting agent for the formation of mitochondrial permeability transition may enhance the toxic neuronal cell injury. PMID- 16715210 TI - Clinical validity of the SWAL-QOL and SWAL-CARE outcome tools with respect to bolus flow measures. AB - The aim of this study was to quantify the association between a dysphagia specific quality of life (SWAL-QOL) and quality of care (SWAL-CARE) questionnaire and four measures of bolus flow. Three hundred eighty-six people with oropharyngeal dysphagia completed a videofluoroscopic examination of their swallowing structure and physiology. They also completed the SWAL-QOL and SWAL CARE surveys. Measures of bolus flow patterns for each swallow were analyzed from videofluoroscopic recordings and correlated with the SWAL-QOL and SWAL-CARE scale scores. The SWAL-QOL and SWAL-CARE scales were modestly related to the four measures of the bolus flow. The SWAL-QOL and SWAL-CARE were most related to measures of oral transit duration and total swallow duration. The SWAL-QOL and SWAL-CARE scales were least related to pharyngeal transit duration. Results were stronger for semisolid trials than for liquid trials. Results were generally weak for the Penetration Aspiration Scale. For all of the significant relationships, the greater the bolus flow severity, the worse the quality of life. The observed modest correlations suggest that patient-centered quality-of-life measures and clinician-driven bolus flow measures provide distinct yet complementary information about oropharyngeal dysphagia. Both sets of measures should be used in dysphagia effectiveness and outcomes research. PMID- 16715211 TI - Initial clinical experience with a partly autonomous robotic surgical instrument server. AB - BACKGROUND: The authors believe it would be useful to have surgical robots capable of some degree of autonomous action in cooperation with the human members of a surgical team. They believe that a starting point for such development would be a system for delivering and retrieving instruments during a surgical procedure. METHODS: The described robot delivers instruments to the surgeon and retrieves the instruments when they are no longer being used. Voice recognition software takes in requests from the surgeon. A mechanical arm with a gripper is used to handle the instruments. Machine-vision cameras locate the instruments after the surgeon puts them down. Artificial intelligence software makes decisions about the best response to the surgeon's requests. RESULTS: A robot was successfully used in surgery for the first time June 16, 2005. The operation involved excision of a benign lipoma. The procedure lasted 31 min, during which time the robot performed 16 instrument deliveries and 13 instrument returns with no significant errors. The average time between verbal request and delivery of an instrument was 12.4 s. CONCLUSIONS: The described robot is capable of delivering instruments to a surgeon at command and can retrieve them independently using machine vision. This robot, termed a "surgical instrument server," represents a new class of information-processing machines that will relieve the operating room team of repetitive tasks and allow the members to focus more attention on the patient. PMID- 16715212 TI - Comparison of laparoscopic and open ileocecal resection for Crohn's disease: a metaanalysis. AB - BACKGROUND: The role of laparoscopic surgery for patients with ileocecal Crohn's disease is a contentious issue. This metaanalysis aimed to compare open resection with laparoscopically assisted resection for ileocecal Crohn's disease. METHODS: A literature search of the Medline, Ovid, Embase, and Cochrane databases was performed to identify comparative studies reporting outcomes for both laparoscopic and open ileocecal resection. Metaanalytical techniques were applied to identify differences in outcomes between the two groups. Sensitivity analysis was undertaken to evaluate the heterogeneity of the study. RESULTS: Of 20 studies identified by literature review, 15 satisfied the criteria for inclusion in the study. These included outcomes for 783 patients, 338 (43.2%) of whom had undergone laparoscopic resection, with an overall conversion rate to open surgery of 6.8%. The operative time was significantly longer in the laparoscopic group, by 29.6 min (p = 0.002), although the blood loss and complications in the two groups were similar. In terms of postoperative recovery, the laparoscopic patients had a significantly shorter time for recovery of their enteric function and a shorter hospital stay, by 2.7 days (p < 0.001). CONCLUSIONS: For selected patients with noncomplicated ileocecal Crohn's disease, laparoscopic resection offered substantial advantages in terms of more rapid resolution of postoperative ileus and shortened hospital stay. There was no increase in complications, as compared with open surgery. The contraindications to laparoscopic approaches for Crohn's disease remain undefined. PMID- 16715213 TI - Metabolic effects of telmisartan in spontaneously hypertensive rats. AB - The favorable metabolic effects of telmisartan are supposedly related to the changes in carbohydrate and lipid metabolism driven by peroxisome proliferators activated receptor-gamma (PPARgamma). The fatty acid translocase CD36 is one of the PPARgamma targets that mediate these actions. We studied the metabolic effects of telmisartan in the NIH-derived strain of spontaneously hypertensive rats (SHR/NIH), which harbors a deletion mutation in CD36, in comparison to the original SHRs (SHR/Izm), which express wild-type CD36. In SHR/Izm, administration of telmisartan was associated with significantly lower serum levels of free fatty acids (42%), triglycerides (29%), glucose (11%), insulin (31%), and lower hepatic triglyceride (17%) levels, as well as larger epididymal fat pads (1.19-fold) than in SHR/NIH. Additionally, insulin-stimulated glucose incorporation into epididymal fat tissues was significantly augmented in SHR/Izm (1.33-fold) compared with SHR/NIH. In the epididymal fat pads of SHR/Izm treated with telmisartan, CD36 mRNA transcript (1.55-fold) and protein expression (1.37-fold) were also significantly enhanced. However, after 4 weeks of treatment with telmisartan, in SHR/NIH only serum free fatty acid levels were slightly reduced (20%). Overall, these results showed marked discrepancies in the metabolic actions of telmisartan in SHR/Izm and SHR/NIH and further supported the involvement of CD36 in the actions of this drug, suggesting that this pharmacogenetic interaction may be of particular importance in CD36-deficient patients. PMID- 16715215 TI - Volumetry of human molars with flat panel-based volume CT in vitro. AB - The flat panel-based volume computed tomography (fpVCT) is a new CT device applicable for experimental, three-dimensional evaluation of teeth at a resolution of about 150 microm in the high contrast region. The aim of this study was to investigate whether fpVCT was suitable for quantification of the volumes of dental hard tissues and the root canal system to establish a new method for morphological studies. Fifty-two extracted third molars (maxillary: 31, mandibular: 21) were examined with a prototype of an fpVCT using a volumetry algorithm at different levels according to the radiographic density of enamel and dentine. Volumetry of the root canal system was performed after "region growing segmentation": starting from a voxel in the centre of the root canal, this algorithm searches voxels of same density in the surrounding. The volumetry of the root canal system was stopped by the investigator at the apical constriction. Results showed that dentine, enamel and root canal system could be well distinguished in three-dimensional images. Volumetry yielded the following data (cm(3), mean+/-SD): dentine 0.438+/-0.111, enamel 0.227+/-0.051, root canal system 0.052+/-0.017 and total volume 0.753+/-0.159. In conclusion, the fpVCT is appropriate for non-destructive volumetry of large numbers of teeth in experimental laboratory studies. PMID- 16715218 TI - Terminal phalangeal accessory ossification center of the thumb: an additional radiographic finding in Larsen syndrome. AB - Larsen syndrome is an autosomal-dominant disorder characterized by multiple joint dislocations, vertebral anomalies and dysmorphic facies. Both autosomal-dominant and autosomal-recessive forms of the disorder have been proposed. Individuals with autosomal-dominant Larsen syndrome have characteristic "cylindrical-shape" thumbs caused by broad, shortened phalanges. Autosomal-dominant Larsen syndrome results from heterozygosity for mutations in filamin B, a cytoskeletal protein involved in multicellular processes. We report here a patient with a duplicated or accessory distal thumb phalanx and multiple large joint dislocations who was shown to be heterozygous for a filamin B mutation predicting the amino acid substitution G1691S. This adds a new radiographic finding, duplicated or accessory distal phalanx, to the radiographic abnormalities seen in this rare dominant disorder. PMID- 16715221 TI - [MRT sequences. Part I]. AB - When magnetic resonance tomography is used in clinical practice a large number of different imaging techniques (sequences) are applied for the imaging. Despite this large number of sequences, which is often difficult to keep track of, all types of sequence work largely according to the same or very similar principles. In this continuing education course these principles are elaborated and the elements that make up a sequence are explained. These elements working together make it possible to assign the signals received from the receiver coil to their point of origin within the body. This first part of the further training is devoted largely to basic observations on spatial encoding of the signals, but also to the difference between 2D and 3D sequences. In addition, the techniques generally used in clinical practice for multislice imaging with 2D images are discussed in some detail. PMID- 16715219 TI - Pediatric DXA: technique and interpretation. AB - This article reviews dual X-ray absorptiometry (DXA) technique and interpretation with emphasis on the considerations unique to pediatrics. Specifically, the use of DXA in children requires the radiologist to be a "clinical pathologist" monitoring the technical aspects of the DXA acquisition, a "statistician" knowledgeable in the concepts of Z-scores and least significant changes, and a "bone specialist" providing the referring clinician a meaningful context for the numeric result generated by DXA. The patient factors that most significantly influence bone mineral density are discussed and are reviewed with respect to available normative databases. The effects the growing skeleton has on the DXA result are also presented. Most important, the need for the radiologist to be actively involved in the technical and interpretive aspects of DXA is stressed. Finally, the diagnosis of osteoporosis should not be made on DXA results alone but should take into account other patient factors. PMID- 16715222 TI - [Differential diagnosis of rheumatic diseases]. AB - QUESTION TO ANSWER: Which imaging modalities are appropriate for the Differential diagnosis of Rheumatic diseases. METHODS AND RESULTS: MRI has far most the highest sensitivity and is unequaled in its brilliant presentation of anatomy and pathology. But it is sometimes forgotten, that this is at least in part the result of carefully selected sequences, dedicated to the expected result. CONCLUSION: Plain X-ray offers high specificity in the differential diagnoses of rheumatic diseases, it is well standardised and it is a device, to use independent from any suspected findings. PMID- 16715223 TI - [Sonography of synovial and erosive inflammatory changes]. AB - High-frequency sonography enables excellent detection of early erosions and synovial proliferations. Power Doppler sonography (PDUS) allows for an improved characterization of articular and peritendinous augmented volume, because detection of hypervascularity correlates with inflammatory activity and further is helpful in differentiation from effusion and inactive pannus. The use of contrast media improves the sensitivity of vascularity detection, because they allow for a delineation of vessels at the microvascular level. This is of increased interest, as the development of new therapeutic options targeting the microvascular level calls for earlier diagnosis and optimal assessment of disease activity. Because of good availability, cost effectiveness, and patient acceptance, sonography facilitates early diagnosis of synovial proliferations and erosions as well as therapy follow-up. PMID- 16715224 TI - [Molecular imaging: future uses in arthritides]. AB - Molecular imaging is an upcoming field in radiology as a result of great advances in imaging technology, genetics, and biochemistry in the recent past. Early-stage imaging of molecular pathological changes in cells opens the gates to new methods in medical treatment of diseases that otherwise would only be detected in advanced stages. Methods of imaging biochemical pathways with molecular agents are currently an issue of intensive research. This article reviews current modalities of molecular imaging in arthritis that should offer future perspective on early disease detection, diagnosis, and monitoring of treatment efficiency and how they can pave the way to optimized therapy. PMID- 16715225 TI - [Quantitative imaging in rheumatoid arthritis: from scoring to measurement]. AB - The need of clinical sciences to measure therapy effects on chronic illness led to development, evaluation, and publication of several radiological methods to monitor disease progression of rheumatic diseases. This review article explains the basics and background of scoring and measurement. The radiologist thus learns to report more compactly and to communicate the results more specifically. PMID- 16715226 TI - [Pancreas. Part II: Tumors]. AB - Adenocarcinoma is the most common malignant pancreatic tumor, affecting the head in 60-70% of cases. By the time of diagnosis, approximately 80% of tumors are unresectable. Helical CT is very effective in detection and staging of adenocarcinoma, with a sensitivity of 76-92% for detection and an accuracy of 80 90% for staging, but it has limitations in the detection of small cancers (< or =2 cm). Multidetector CT (MDCT) has brought substantial improvements with its inherent 3D imaging capability. Mangafodipir-enhanced MRI is a problem-solving tool in the depiction of small cancers following an equivocal CT imaging result. Gadolinium-enhanced 3D gradient-echo MRI is helpful in the assessment of vascular invasion of cancer and in determining the etiology of cystic lesions. Serous cystadenoma is benign, has a lobulated contour and contains innumerable small cysts of 0.1-2 cm in diameter. Mucinous cystic neoplasms are unilocular or multilocular (fewer than six cysts), and the cyst diameter exceeds 2 cm. The presence of solid nodular components should alert the radiologist to suspect cystadenocarcinoma. Neuroendocrine tumors are mostly hypervascular. Diagnosis of insulinoma is a challenge: they are <2 cm in 90% of cases and mostly hypervascular at CT or MRI. A combination of contrast-enhanced MDCT, MRI, endosonography, and/or somatostatin receptor scintigraphy is used to detect these small tumors. This review summarizes the imaging features of the most common pancreatic tumors and discusses the limitations of CT, MRI and endosonography. PMID- 16715228 TI - Extramammary myofibroblastoma is genetically related to spindle cell lipoma. AB - Extramammary-type myofibroblastoma is a rare, benign spindle cell lesion, strictly resembling the breast counterpart, but occurring in extramammary sites, mainly in the inguinal/groin area. In this paper, we describe an extramammary type myofibroblastoma in the groin of a 37-year-old male patient. The tumor showed a typical morphological and immunophenotypical profile, including staining for both CD34 and desmin. Dual-color interphase florescent in situ hybridization analysis revealed losses of RB/13q14 and FKHR/13q14 loci within tumor cells. The chromosome 13 rearrangements associated with the loss of the 13q14 chromosomal region are typically seen in spindle cell lipoma, and have been previously recognized in mammary myofibroblastoma, providing strong evidence for a pathogenetic link between these lesions. PMID- 16715229 TI - Genetic clonality is a feature unifying nephroblastomas regardless of the variety of morphological subtypes. AB - Nephroblastomas are embryonal tumors exhibiting a wide variety of different morphological features and genetic changes. Some of the genetic aberrations were associated with a certain histological subtype. It is generally assumed that nephroblastomas develop as subclonal proliferations from nephrogenic rests. However, so far, a very limited amount of tumors from only part of the morphological spectrum of nephroblastomas was investigated. We therefore investigated the clonality of 45 tumors of all different histological subtypes. The number of each subtype was in accordance with the percentage of occurrence of the respective subtype. We analyzed a highly polymorphic locus of the human androgen receptor gene for nonrandom X-inactivation of genomic DNA using a methylation-sensitive restriction enzyme. Data were obtained for 39 tumors. Eighteen of the tumors included were noninformative in the genetic locus examined, the remaining 21 tumors were monoclonal regardless of the histological subtype. Our findings therefore support the hypothesis that Wilms' tumors are monoclonal proliferations despite their large variety of morphological features. PMID- 16715230 TI - An unusual case of malignant pilocytic astrocytoma occurring in the eye. AB - Pilocytic astrocytoma is a central nervous system neoplasia that arises during pediatric age. Only few cases have been documented in patients older than 50 years old. It is a low-grade lesion that can rarely undergo malignant changes presenting the histologic features of a high-grade glioma. We report a case of a pilocytic astrocytoma arising in the eyeball of a 53-year-old man affected by glaucoma that underwent malignant evolution. PMID- 16715232 TI - Hydrolysis of black soybean isoflavone glycosides by Bacillus subtilis natto. AB - Hydrolysis of isoflavone glycosides by Bacillus subtilis natto NTU-18 in black soymilk is reported. At the concentration of 3-5% (w/v), black soymilk in flask cultures, the isoflavones, daidzin, and genistin were highly deglycosylated within 24 h. Deglycosylation of isoflavones was further carried out in a 7-l fermenter with 5% black soymilk. During the fermentation, viable cells increased from 10(3) to 10(9) CFU ml(-1) in 15 h, and the activity of beta-glucosidase appeared at 8 h after inoculation and reached a maximum (3.3 U/ml) at 12 h, then decreased rapidly. Deglycosylation of isoflavone glycosides was observed at the same period, the deglycosylation rate of daidzin and genistin at 24 h was 100 and 75%, respectively. It is significantly higher than the previous reports of fermentation with lactic acid bacteria. In accordance with the deglycosylation of isoflavone glycosides, the estrogenic activity of the 24 h fermented black soymilk for ERbeta estrogen receptor increased to threefold; meanwhile, the fermented broth activated ERalpha estrogen receptor to a less extent than ERbeta. These results suggest that this fermentation effectively hydrolyzed the glycosides from isoflavone in black soymilk and the fermented black soymilk has the potential to be applied to selective estrogen receptor modulator products. PMID- 16715231 TI - The cardiac isoform of alpha-actin in regenerating and atrophic skeletal muscle, myopathies and rhabdomyomatous tumors: an immunohistochemical study using monoclonal antibodies. AB - The two sarcomeric isoforms of actins, cardiac and skeletal muscle alpha-actin, are highly homologous so that their immunohistochemical distinction is extremely difficult. Taking advantage of monoclonal antibodies distinguishing the two conservative amino acid exchanges near the aminoterminus, we have performed an extended immunohistochemical analysis of the cardiac alpha-actin (CAA) isoform in normal, regenerating, diseased and neoplastic human muscle tissues. Intense and uniform CAA staining is seen in fetal and adult myocardium and in fetal skeletal muscle while adult skeletal muscle is essentially negative, except for muscle spindle myocytes and a few scattered muscle fibres with overall reduced diameter. By contrast, CAA synthesis is markedly induced in regenerating skeletal muscle cells, in Duchenne muscular dystrophy and upon degenerative atrophy. CAA has also been detected in certain vascular and visceral smooth muscle cells. Among tumors, CAA has consistently been seen in rhabdomyosarcomas and rhabdomyomatous cells of nephroblastomas, whereas, smooth muscle tumors have shown only occasional staining. While the synthesis of this actin isoform is less restricted than previously thought, monoclonal antibodies against CAA provide a well-defined, reliable and sensitive diagnostic tool for the definition and detection of aberrant differentiation in diseased skeletal muscle and of striated muscle differentiation in rhabdomyosarcomas. PMID- 16715233 TI - Distribution of muscle larvae and antibody dynamics in goats experimentally infected with Trichinella spiralis. AB - Herbivorous animals can play a very important role in spreading trichinellosis. In the study presented here, the susceptibility and distribution of Trichinella spiralis infection was examined in 16 goat kids. The goats were inoculated with 10,000 T. spiralis larvae isolated by artificial digestion methods. The animals were necropsied per two animals in weekly intervals, and the larval burdens in different muscle tissue and anti-Trichinella antibodies measured with the indirect enzyme-linked immunosorbent assay (ELISA) serological method using excretory-secretory (E/S) antigen for detecting anti-Trichinella antibodies were assessed during the experiment. T. spiralis larval burden was maximal at 6 weeks postinoculation (480-5,057 larvae/g according to locality), and the larvae were also found in the myocardium (0.77 larvae/g). In this paper, our next step was to compare the specificity and the time of seroconversion by means of ELISA based on E/S antigen prepared from T. spiralis. Antibody response was detected in all 16 goats. The ELISA test carried out showed the first increments in optical density 2 weeks postinfection (p.i.), reached their peak 4 weeks p.i., and remained elevated from that day until the end of the experiment (10 weeks p.i.). These results indicated that specific anti-Trichinella antibodies in goats persist for a relatively long time. PMID- 16715234 TI - Prevalence of helminthes in adult dogs in Heilongjiang Province, the People's Republic of China. AB - The prevalence of helminthes in adult dogs was investigated in Heilongjiang Province, the People's Republic of China, between 1996 and 2004. A total of 178 adult farm dogs from representative geographical locations in Heilongjiang Province were killed and examined for the presence of helminthes using a helminthological approach. The worms were examined, counted, and identified to species according to existing keys and descriptions. A total of 17 species of helminthes were found to infect dogs, and they represented two phyla, three orders, 13 families, and 15 genera. All dogs were infected by more than one helminth species. Clonorchis sinensis (26.4%), Paragonimus westermani (7.9%), and Metagonimus yokogawai (6.2%) were the most common trematode species; Mesocestoides lineatus (20.2%), Taenia hydatigena (19.7%), and Dipylidium caninum (14.6%) were the most common cestodes species; and Ancylostoma caninum (66.3%), Toxocara canis (36.5%), and Trichinella nativa (21.9%) were the most common nematode species. The results of the present investigation provide relevant "base line" data for assessing the effectiveness of future control strategies against helminth infection in dogs in Heilongjiang Province, China. PMID- 16715235 TI - Seroprevalence of canine babesiosis in Hungary suggesting breed predisposition. AB - Six hundred fifty-one blood samples were collected from urban and rural dogs in various parts of Hungary to measure antibody levels to Babesia canis with indirect fluorescent antibody test. Thirty-seven (5.7%) of the sera showed positivity with titers between 1:80 and 1:10,240. Seroconverted dogs were found in 13 locations of the country. It is concluded that canine babesiosis is becoming more prevalent in Eastern Hungary. Seropositivity increased then declined with age, reaching a maximum in case of 3.1- to 5-year-old dogs. Prevalence of antibodies to B. canis was significantly higher among german shepherds and komondors. This suggests a genetic predisposition of german shepherd dogs to chronic babesiosis (carrier status) with long-term maintenance of their seropositivity. On the other hand, heavy-coated komondors are phenotypically more suitable for repeated exposure to ticks, potentially infected with B. canis. This is the first report on the seroprevalence of canine babesiosis in Hungary. PMID- 16715236 TI - Scientific papers presented at the 2000-2001 European Society of Gastrointestinal and Abdominal Radiology (ESGAR) meetings: publication rates during the period 2000-2004. AB - The aim of this study was to investigate the rate at which abstracts orally presented at the ESGAR 2000 and 2001 meetings were published between 2000-2004, and to identify predictive factors of publication. The abstracts of ESGAR meeting presentations were reviewed and classified according to organ, modality, type of design, country of origin of the studies and basic categories of diagnostic or interventional. The presentations were searched for publication in Medline indexed journals using the PubMed server. The publication rates of the presentations, the time period between the presentation and publication, and the journal in which the article had appeared were investigated. An overall number of 109 publications were found originating from 276 presentations (39.5%). The median and inter-quartile range [IQR] between the abstract presentation and subsequently full publication was 18 months [1.0 -53.0]. The journal with the highest number of derived articles from abstract presentation was European Radiology (n=21, 19.2%). Retrospective studies were found to be more frequently published than prospective studies (p=0.001). The publication rate did not show any statistically significant difference between groups of other classifications. The publication rate of studies orally presented at ESGAR meetings was 39.5%. No specific variable other than the type of design of the studies appeared to influence the publication rate. PMID- 16715237 TI - Cardiac stress MR imaging with dobutamine. AB - Stress testing for detection of ischemia-induced wall-motion abnormalities has become a mainstay for noninvasive diagnosis and risk stratification of patients with suspected coronary artery disease (CAD). Recent technical developments in magnetic resonance imaging (MRI), including the adoption of balanced steady-state free precession (b-SSFP) sequences-preferentially in combination with parallel imaging techniques-have led to a significant reduction of imaging time and improved patient safety. The stress protocol includes application of high-dose dobutamine (up to 40 microg/kg/min) combined with fractionated atropine (up to a maximal dose of 1.0 mg). High-dose dobutamine stress MRI revealed good sensitivity (83-96%) and specificity (80-100%) for detection of significant CAD. Myocardial tagging methods have been shown to further increase sensitivity for CAD detection. Severe complications (sustained tachycardia, ventricular fibrillation, myocardial infarction, cardiogenic shock) are rare but may be expected in 0.1-0.3% of patients. Dobutamine stress MRI has emerged as a reliable and safe clinical alternative for noninvasive assessment of CAD. New pulse sequences, such as real-time imaging, might obviate the need for breath holding and electrocardiogram (ECG) triggering in patients with severe dyspnoea and cardiac arrhythmias, which may further improve the clinical impact and acceptance of stress MRI in the future. PMID- 16715238 TI - Imaging in percutaneous ablation for atrial fibrillation. AB - Percutaneous ablation for electrical disconnection of the arrhythmogenic foci using various forms of energy has become a well-established technique for treating atrial fibrillation (AF). Success rate in preventing recurrence of AF episodes is high although associated with a significant incidence of pulmonary vein (PV) stenosis and other rare complications. Clinical workup of AF patients includes imaging before and after ablative treatment using different noninvasive and invasive techniques such as conventional angiography, transoesophageal and intracardiac echocardiography, computed tomography (CT) and magnetic resonance imaging (MRI), which offer different information with variable diagnostic accuracy. Evaluation before percutaneous ablation involves assessment of PVs (PV pattern, branching pattern, orientation and ostial size) to facilitate position and size of catheters and reduce procedure time as well as examining the left atrium (presence of thrombi, dimensions and volumes). Imaging after the percutaneous ablation is important for assessment of overall success of the procedure and revealing potential complications. Therefore, imaging methods enable depiction of PVs and the anatomy of surrounding structures essential for preprocedural management and early detection of PV stenosis and other ablation related procedures, as well as long-term follow-up of these patients. PMID- 16715239 TI - PET-positive fibrous dysplasia--a potentially misleading incidental finding in a patient with intimal sarcoma of the pulmonary artery. AB - Benign bone tumors can show an increased FDG uptake in FDG-PET/CT investigations. In the presented case, an incidentally detected PET-positive asymptomatic fibrous dysplasia was initially misinterpreted as a metastasis in a patient with intimal sarcoma of the pulmonary artery. PMID- 16715240 TI - Acetabular retroversion as a rare cause of chronic hip pain: recognition of the "figure-eight" sign. AB - While well-recognized in the orthopedic literature as a cause of chronic hip pain, acetabular retroversion has not been specifically described in the radiologic literature. Acetabular retroversion represents a particular form of hip dysplasia characterized by abnormal posterolateral orientation of the acetabulum. This pathophysiology predisposes the individual to subsequent anterior impingement of the femoral neck upon the anterior acetabular margin and fibrous labrum. Without treatment, cases may progress to damage of the anterior labrum and cartilage, with eventual early onset of osteoarthritic disease. This impinging condition has been described as occurring in isolation or as part of a complex dysplasia. We describe two cases of acetabular retroversion diagnosed by conventional radiographic evaluation of the pelvis, one in isolation and one occurring in the setting of a larger congenital syndrome. These cases illustrate the utility of the "figure-eight" sign in identifying abnormalities of acetabular version and thus assisting clinicians in properly identifying these individuals so that appropriate therapy may be instituted. PMID- 16715241 TI - Recurrent acute low back pain secondary to lumbar epidural calcification. AB - INTRODUCTION: Epidural calcification is a rare cause of back pain, and spontaneous epidural calcification has not been reported previously. CASE REPORT: We describe a patient with acute low back pain and signs of lumbar nerve root compression due to epidural calcification, as demonstrated by CT-scan and MRI. Radiological signs of spondylodiscitis led to a search for an infectious cause, which was negative, and her symptoms responded rapidly to NSAID treatment alone. Her symptoms recurred 18 months later, and further imaging studies again revealed epidural calcification, but with a changed distribution. Her symptoms were relieved once more by NSAID treatment alone. DISCUSSION: We propose that epidural calcification secondary to aseptic spondylodiscitis is the main cause of acute back pain in this patient. A possible mechanism may be the pro-inflammatory effects of calcium pyrophosphate or hydroxyapatite crystal deposition within the epidural space. PMID- 16715242 TI - Deep and superficial infrapatellar bursae: cadaveric investigation of regional anatomy using magnetic resonance after ultrasound-guided bursography. AB - OBJECTIVE: To demonstrate the anatomy of the deep and superficial infrapatellar bursae using magnetic resonance (MR) imaging and anatomic correlation in cadavers. DESIGN: MR imaging of the infrapatellar bursae of nine cadaveric knees was performed after ultrasound-guided bursography. The images were compared with those seen on anatomic sectioning. Histologic analysis was obtained in two specimens. RESULTS: The deep infrapatellar bursa (DIB) was visualized in all specimens (n=9) and the superficial infrapatellar bursa (SIB) in five specimens (55%). The mean dimensions of the DIB in the craniocaudal, mediolateral, and anteroposterior planes were respectively 25, 28.7, and 6 mm, and for SIB 19.5, 21.2 and 2.2 mm. A fat apron dividing the DIB was depicted in eight knees (89%). Lateral extension of the DIB beyond the patellar tendon was observed in 100% of cases. Cadaveric analysis depicted a thin septum in the SIB in four of five cases (80%). CONCLUSIONS: The DIB is generally present and extends beyond the lateral margin of the patellar tendon. A fat apron partially separating this structure is usual. The SIB is not an unusual finding and may have a septum separating its compartments. PMID- 16715243 TI - Cerebrotendinous xanthomatosis presenting with bilateral Achilles tendon xanthomata. AB - We report on a case of a 36-year-old lady who presented with large, painful soft tissue swellings of both Achilles tendons. MRI demonstrated fusiform enlargement involving the Achilles tendons bilaterally. The tendons returned heterogeneous signal intensity characterised by a diffuse reticulated appearance. The right tendon mass was treated with a wide marginal excision and Achilles tendon reconstruction. The histology confirmed Achilles tendon xanthoma. Further metabolic investigation revealed the patient to have a rare autosomal recessive condition called cerebrotendinous xanthomatosis (CTX). Her brother was also affected. CTX is easily treatable if diagnosed early, and should be suspected in patients presenting with bilateral Achilles tendon xanthomas and normal plasma lipid levels. PMID- 16715244 TI - Transverse morphology of the sacroiliac joint: effect of angulation and implications for fluoroscopically guided sacroiliac joint injection. AB - OBJECTIVE: Effects of angulation of computed tomography (CT) reconstruction plane on sacroiliac (SI) joint morphology were studied, and factors influencing the approach to fluoroscopically guided SI joint injection were assessed. DESIGN AND PATIENTS: CT scans of pelvises were reformatted on 41 subjects, aged 51.7 (+/ 15.1) years. Transverse images were reconstructed at the caudal 3 cm of the SI joint tilting plane of reconstruction from -30 degrees to +30 degrees at 15 degrees increments. Anteroposterior diameter of joint (depth), angle from sagittal plane (orientation angle), and distance from skin were measured. Joint contour was classified, and presence of bone blocking access to the joint was recorded. Comparison between angles were analysed by t-test. Relationships between variables were assessed by a Pearson correlation test. RESULTS: Depth was shorter with angulation in the inferior direction (P<0.01). Orientation angle increased with superior angulation (P<0.01). Distance from skin increased (P<0.01) with angulation in either direction. Joint contour was significantly different from baseline at each angle (P<0.001) but highly variable. Inferior angulation resulted in interposition of ilium between skin and SI joint, and superior angulation caused bone block due to the lower sacrum. None of these features was identified without tilting of the reconstruction plane, and effects were more pronounced with steeper angulation. CONCLUSION: Angulation of the reconstruction plane considerably affects the appearance of the sacroiliac joints. By shortening joint depth, an inferiorly directed approach to SI joint injection may make fluoroscopic guidance easier, although associated bony interposition can prevent access to the synovial compartment. A superiorly directed approach is more likely to have adverse effects. PMID- 16715245 TI - Spondyloarthropathy presenting at a young age: case report and review. AB - The diagnosis of juvenile spondyloarthritis (JSA) is rarely entertained in young children who present with back and leg pain. We present a case of a 6-year-old male who presented with a 3-year history of severe back and leg pain and a positive Gower's sign, and was given a presumed diagnosis of muscular dystrophy. Presenting serologic evaluation included a mildly elevated sedimentation rate and C-reactive protein (CRP). Computed tomography of the pelvis demonstrated large erosions affecting both sacro-iliac joints. Despite the unusually young age of this patient, ankylosing spondylitis seemed the most plausible diagnosis. Following rheumatological evaluation and treatment for JSA, he showed significant clinical improvement. His disease, however, has not entirely remitted with signs of enthesitis at the Achilles tendon and knees. We present this case to illustrate that JSA could account for symptoms at an early age and not considering it could lead to multiple medical visits and diagnoses. To our knowledge, based on a search of the World literature, this would appear to be the youngest case of JSA reported with demonstrable severe sacroiliitis. PMID- 16715246 TI - Comments on 'Is partial coherence a viable technique for identifying generators of neural oscillations?': Why the term 'Gersch Causality' is inappropriate: common neural structure inference pitfalls. AB - To aid prospective neural connectivity inference analysts and hoping to preclude misconception spread, we exploit the didatic value of some of the issues raised by Albo et al. (Biol Cybern 90: 318-326, 2004) who claim that signal-to-noise ratio (SNR) values can lead to mistakes in structural inference when using partial coherence in connection to Gersch's 1970 method for spotting signal sources (Gersch in Math Biosci 14: 177- 196, 1972). We show theoretically that Gersch's method is able only to spot which measurement of some common underlying factor has the least amount of additive noise and that this has nothing to do with any reasonable notion of 'causality' as suggested by Albo et al. (Biol Cybern 90: 318-326, 2004). We also show that despite the inherent structural ambiguity of the model used by Albo et al. (Biol Cybern 90: 318-326, 2004) to back their claim, its data can nonetheless furnish the correct time precedence hierarchy between the activities in its measured structures, both when simple (correlation) and more sophisticated methods are used (partial directed coherence) (Baccala and Sameshima in Biol Cybern 84:463-474, 2001a) in a true depiction of time series causality. PMID- 16715249 TI - Pregnancy and delivery in patients with enterostomy due to anorectal complications from Crohn's disease. AB - BACKGROUND AND AIMS: Enterostomy is often undergone by patients with Crohn's disease (CD) due to severe anorectal lesions such as rectovaginal fistula (RVF). Reports of successful pregnancy and delivery, which are important determinants of quality of life for female CD patients with stoma, are limited. Thus, we investigated problems associated with pregnancy and delivery in female CD patients at our hospital. MATERIALS AND METHODS: Between 1985 and 2003, five female CD patients with enterostoma carried seven pregnancies and delivered eight babies in our hospital. For this study, we investigated CD activity, fetal growth, stoma complications, and the outcome of delivery in these seven pregnancies. RESULTS: Among the five patients, four underwent loop ileostomy and one loop sigmoidostomy for treatment of RVF or severe stricture of the rectum or sigmoid colon. Except for one case, no fertility treatment was done. During pregnancy, a flare-up was observed in one patient and was successfully treated with corticosteroids. Although home enteral or parenteral nutrition was required in two cases, fetal growth was within the normal range in all pregnancies. Preterm delivery occurred in one case with a twin pregnancy. The mean diameter of stoma increased during pregnancy (p<0.01), and a mucosal laceration of stoma by the edge of an ostomy appliance occurred in one case. All deliveries were safe, with six cesarean sections and one transvaginal delivery. After each delivery, the stoma returned to pre-pregnancy size. CONCLUSIONS: Pregnancy and delivery in CD patients with stoma is safe and should be encouraged. However, special attention to disease activity, nutritional support, and stoma-related complications is recommended. PMID- 16715250 TI - (E)-metanicotine hemigalactarate (TC-2403-12) inhibits IL-8 production in cells of the inflamed mucosa. AB - BACKGROUND: Nicotine is of therapeutic value in ulcerative colitis, but its administration is connected with adverse events. Nicotine derivatives are currently being tested to maintain the therapeutic effects and minimize adverse events. TC-2403-12 is a (E)-metanicotine hemigalactarate. The aim of this study was to determine the effectiveness of TC-2403-12 in the inhibition of TNF- and lipopolysaccharide (LPS)-induced cell activation. METHODS: Colonic epithelial cells (CEC), monocytes (MM6), granulocytes, and the intestinal epithelial cell line HT-29 were stimulated with TNF and LPS and treated with TC-2403-12. IL-8 secretion in the cell supernatants and NF-kappaB activation were determined by ELISA. Apoptosis was quantified by flow cytometry. RESULTS: In MM6 cells, IL-8 secretion was significantly decreased to 30% of control after TC-2403-12 treatment, with best results after pretreatment for 24 h. This decrease in cell activation was not due to apoptosis and was not mediated by inhibition of NF kappaB activation. IL-8 production in neutrophils and primary CEC also tended to be decreased after TC-2403-12 treatment. TC-2403-12 had no influence on IL-8 secretion of HT-29 cells. CONCLUSION: TC-2403-12 effectively inhibited TNF- and LPS-induced IL-8 production in different cell types. No toxic effects occurred at the concentrations used. Preincubation of cells with TC-2403-12 showed the best effects. PMID- 16715252 TI - Penetration of topically applied levofloxacin 0.5% and ofloxacin 0.3% into the vitreous of the non-inflamed human eye. AB - BACKGROUND: The aim of the study was to evaluate the vitreous penetration of two commercially available ophthalmic fluoroquinolones: ofloxacin and levofloxacin. METHODS: This prospective, double-blind, randomized clinical trial comprised 16 patients scheduled for vitrectomy surgery of one eye for macular hole or macular pucker. The patients were randomly assigned to receive topical ofloxacin 0.3% (n=9) or levofloxacin 0.5% (n=7) the day before, one drop at noon, 4 p.m., 8 p.m. and midnight. The next morning, patients were given their assigned masked antibiotic every 5 min for four doses starting 1 h before surgery. The vitreous humour samples, at least 0.3 ml each, were collected 1 h after the administration of the last dose, at the beginning of the pars plana vitrectomy with infusion disconnected. Samples were assayed for ofloxacin and levofloxacin concentrations by a method using high-performance liquid chromatography (HPLC) coupled with single mass spectrometry with electrospray ionization RESULTS: Equal topical administration of levofloxacin yielded 2.5 times higher vitreal concentration than ofloxacin. The mean vitreous concentrations of ofloxacin and levofloxacin were 5.30+/-3.04 (SD) ng/ml and 13.09+/-5.24 ng/ml, respectively (P=0.002). CONCLUSIONS: Equal dosing with topical administration of levofloxacin 0.5% and ofloxacin 0.3% allows better penetration into the vitreous for levofloxacin, but the levels of mean concentrations of each drug did not exceed the MIC(90) or MIC(50) for most ocular pathogenic bacteria in terms of conventional endophthalmitis therapy. PMID- 16715253 TI - Carboxymethylcellulose as a new carrier substance for intravitreal injection of reproducible amounts of triamcinolone. AB - BACKGROUND: Intravitreal application of triamcinolone acetonide has become increasingly popular for the treatment of various retinal disorders. However, dosage, mode of preparation and application differ worldwide. The aim of this study was to find a safe vehicle that would allow intravitreal injection of an exact amount of triamcinolone acetonide without potentially retinotoxic preservatives. METHODS: Solutions of triamcinolone acetonide with a theoretical concentration of 4 mg/0.2 ml were prepared following one sedimentation (A) and two filtration (B, C) methods. In addition, a filtration method using carboxymethylcellulose 2% as a carrier (D) was established. During processing and after injection into an eye model, the crystals were quantified by weight and high-performance liquid chromatography (HPLC), and, hence, the rate of crystal loss during this process was determined. RESULTS: The initial preparation contained 93-106% of the calculated quantity. Method A, containing the entire vehicle, delivered 45%+/-7.3% of the target quantity to the eye model, whereas the vehicle-free methods B and C delivered 15%+/-6.9% and 11%+/-3.2%, respectively. Using carboxymethylcellulose 2% as a preservative-free vehicle, we found 93%+/-3.7% of the calculated amount in the eye model. The missing crystals were mainly sticking to the walls of the syringes and needles used for transfer. CONCLUSION: Common methods for preparing triamcinolone acetonide vary in the amount of drug actually injected intravitreally. Carboxymethylcellulose is an ideal carrier substance for intravitreal application of an exact dose of triamcinolone acetonide without preservatives. PMID- 16715254 TI - Regulatory T cells: present facts and future hopes. AB - Naturally occurring CD4(+)CD25(+)Foxp3(+) regulatory T cells and several subsets of induced suppressor T cells are key players of the immune tolerance network and control the induction and effector phase of our immunological defense system. These T cell populations actively control the properties of other immune cells by suppressing their functional activity to prevent autoimmunity and transplant rejection but also influence the immune response to allergens as well as against tumor cells and pathogens. Even though we are far from completely understanding the molecular and cellular mechanisms that manage the different regulatory T cell populations, increasing evidence exists about their functional importance. The knowledge on their induction and activation opens the possibility for their selective manipulation in vivo as an attractive approach for an immunotherapy of unwanted immune responses. This review summarizes this knowledge and discusses the potential of regulatory T cells for novel immunointervention strategies in the future. PMID- 16715255 TI - Ventral hippocampal alpha7 and alpha4beta2 nicotinic receptor blockade and clozapine effects on memory in female rats. AB - RATIONALE: Nicotinic systems in the hippocampus play important roles in memory function. Decreased hippocampal nicotinic receptor concentration is associated with cognitive impairment in schizophrenia and Alzheimer's disease. METHODS: We modeled in rats the cognitive effects of chronic decrease in hippocampal alpha7 or alpha4beta2 receptors with 4-week continuous bilateral local infusions of the alpha7 nicotinic antagonist methyllycaconitine (MLA) or the alpha4beta2 antagonist dihydro-beta-erythroidine (DHbetaE). The working memory effects of these infusions were assessed by performance on the radial-arm maze. To test the effect of antipsychotic medication, we gave acute injections of clozapine and to determine the impact of nicotine, which is widely used by people with schizophrenia approximately half of the rats received chronic systemic infusions of nicotine. RESULTS: Chronic ventral hippocampal DHbetaE infusion caused a significant (p < 0.001) working memory impairment. Acute systemic clozapine (2.5 mg/kg) caused a significant (p < 0.005) working memory impairment in rats given control aCSF hippocampal infusions. Clozapine significantly (p < 0.025) attenuated the memory deficit caused by chronic hippocampal DHbetaE infusions. Chronic ventral hippocampal infusions with MLA did not significantly affect the working memory performance in the radial-arm maze, but it did significantly (p < 0.05) potentiate the memory impairment caused by 1.25 mg/kg of clozapine. Chronic systemic nicotine did not significantly interact with these effects. CONCLUSIONS: The state of nicotinic receptor activation in the ventral hippocampus significantly affected the impact of clozapine on working memory with blockade of alpha7 nicotinic receptors potentiating clozapine-induced memory impairment and blockade of alpha4beta2 receptors reversing the clozapine effect from impairing to improving memory. PMID- 16715263 TI - Longitudinal diffusion behavior of hemicyanine dyes across phospholipid vesicle membranes as studied by second-harmonic generation and fluorescence spectroscopies. AB - The adsorption and longitudinal diffusion behaviors of a series of hemicyanine dyes to phospholipid vesicle membranes were studied by second-harmonic generation (SHG) and fluorescence spectroscopies. It was observed that the longitudinal diffusion of cationic hemicyanine dyes takes place immediately after the initial adsorption of these dyes to the outer surface of the vesicle membrane. In contrast, hardly any amount of a zwitterionic hemicyanine dye with a sulfonate group diffused across the vesicle membrane within the measurement time (<2000 s). Based on the difference in the time-course responses of SHG and fluorescence spectroscopies for all of the hemicyanine dyes tested, we propose that hydration of the sulfonate group is mainly responsible for the low diffusivity of the zwitterionic hemicyanine dye. PMID- 16715268 TI - Good oral presentation of scientific work. PMID- 16715266 TI - Environmental fate processes and biochemical transformations of chiral emerging organic pollutants. AB - This review highlights the analytical chemistry, environmental occurrence, and environmental fate of individual stereoisomers of chiral emerging pollutants, which are modern current-use chemicals of growing environmental concern due to their presence in the environment and potential for deleterious effects. Comparatively little is known about individual stereoisomers of pollutants, which can have differential toxicological effects and can be tracers of biochemical weathering in the environment. Stereoisomers are resolved by gas chromatography (GC), high-performance liquid chromatography (HPLC), and capillary electrophoresis (CE). Separation techniques in environmental analysis are typically coupled to mass spectrometry (MS) and tandem mass spectrometry (MS/MS), as these provide the sensitivity and selectivity needed. The enantiomer composition of phenoxyalkanoic and acetamide herbicides, organophosphorus and pyrethroid pesticides, chiral polychlorinated biphenyl metabolites, synthetic musks, hexabromocyclododecane, and pharmaceuticals in the environment show species-dependent enantioselectivity from biotransformation and other biologically mediated processes affecting enantiomers differentially. These enantiomer compositions are useful in detecting biologically mediated environmental reactions, apportioning sources of pollutants, and gaining insight into the biochemical fate of chiral pollutants in the environment, which are needed for accurate risk assessment of such chemicals. PMID- 16715270 TI - Cell trapping in microfluidic chips. PMID- 16715256 TI - An operant determination of the behavioral mechanism of benzodiazepine enhancement of food intake. AB - RATIONALE: A recent review paper by Cooper (Appetite 44:133-150, 2005) has pointed out that a role for benzodiazepines as appetite stimulants has been largely overlooked. Cooper's review cited several studies that suggested the putative mechanism of enhancement of food intake after benzodiazepine administration might involve increasing the perceived pleasantness of food (palatability). OBJECTIVES: The present study examined the behavioral mechanism of increased food intake after benzodiazepine administration. MATERIALS AND METHODS: The cyclic-ratio operant schedule has been proposed as a useful behavioral assay for differentiating palatability from regulatory effects on food intake (Ettinger and Staddon, Physiol Behav 29:455-458, 1982 and Behav Neurosci 97:639-653, 1983). The current study employed the cyclic-ratio schedule to determine whether the effects on food intake of chlordiazepoxide (CDP) (5.0 mg/kg), sodium pentobarbital (5.0 mg/kg), and picrotoxin (1.0 mg/kg) were mediated through palatability or regulatory processes. RESULTS: The results of this study show that both the benzodiazepine CDP and the barbiturate sodium pentobarbital increased food intake in a manner similar to increasing the palatability of the ingestant, and picrotoxin decreased food intake in a manner similar to decreasing the palatability of the ingestant. CONCLUSIONS: These results suggest that the food intake enhancement properties of benzodiazepines are mediated through a mechanism affecting perceived palatability. PMID- 16715271 TI - Hair analysis: an excellent tool for confirmation of drug abuse. PMID- 16715272 TI - Integrating chemical synthesis and analysis on a chip. PMID- 16715273 TI - Superheated water: the ultimate green solvent for separation science. PMID- 16715274 TI - Immunoassay development for environmental analysis. PMID- 16715275 TI - FloDots: luminescent nanoparticles. AB - Luminescent dye-doped silica nanoparticles (FloDots) have been developed for ultrasensitive bioanalysis and diagnosis in the past several years. Those novel nanoparticles are highly luminescent and extremely photostable. In this paper, we review the preparation, characterization, bioconjugation and bioapplication of FloDots. All the results clearly demonstrated that FloDots have many advantages over currently used luminescent probes, such as traditional fluorophores and quantum dots. PMID- 16715276 TI - Applications of the luminol chemiluminescent reaction in analytical chemistry. AB - This critical review discusses the results published between 2000 and 2005 on the development of analytical systems based on the luminol chemiluminescent and electrochemiluminescent reactions. An increasing number of non-specific detection systems based on the enhancing, inhibiting or catalysing effect of a large range of compounds have been published. Possible detected compounds and their concomitant presence in samples are discussed. Chemiluminescent and electrochemiluminescent reactions were also found to merge in biochip and microarray development as a possible substitute to the well-established but hardly quantitative fluorescent detections. PMID- 16715277 TI - Speciation of alkylated metals and metalloids in the environment. AB - The analytical methodology for speciation of metals and metalloids associated with alkyl groups and biomacromolecules is critically reviewed. Alkylated metals and metalloids are not only known to be produced by microbial methylation within most anaerobic compartments in the environment, but also in the course of enzymatic transformations during human metabolism. Because of the toxicological relevance of these compounds present in trace to ultratrace concentrations, firm species identification and exact quantification are essential. While many instrumental techniques coupling chromatography (GC, HPLC, CE, GE) with plasma mass spectrometry (ICP-MS) are available for quantification, methods used for structural identification often suffer from inadequate sensitivity (EI-MS, ESI MS, MALDI-MS, FT-ICRMS). Other problems encountered are sample derivatisation artefacts, lack of suitable standards for quantification, lack of equilibrium between spikes and sample, and the integrity of metal-protein association during separation, in particular during SDS-PAGE. Selected application examples with respect to mercury and arsenic speciation will be discussed critically. PMID- 16715278 TI - Determination of isopropylthioxanthone (ITX) in milk, yoghurt and fat by HPTLC FLD, HPTLC-ESI/MS and HPTLC-DART/MS. AB - Two new HPTLC methods for quantification of isopropyl-9H-thioxanthen-9-one (ITX) in milk, yoghurt and fat samples have been developed. Extraction of ITX from milk and yoghurt was performed with a mixture of cyclohexane and ethyl acetate by employment of accelerated solvent extraction (ASE). For soy bean oil and margarine, a simple partitioning of ITX into acetonitrile was used. ITX and 2,4 diethyl-9H-thioxanthen-9-one (DTX) used as internal standard have been separated on silica gel 60 HPTLC plates with a mixture of toluene and n-hexane (4:1, v/v) and on RP18 HPTLC plates with a mixture of acetonitrile and water (9:1, v/v). Development was performed anti-parallel from both plate sides leading to a throughput of 36 separations in 7 min. Fluorescence measurement at 254/>400 nm was used for quantification. Limits of detection (S/N of 3) have been established to be 64 pg for ITX and DTX on both types of HPTLC plates. In fatty matrix (spiked butter) LOD of ITX was determined to be 1 mug kg(-1). In the working range monitored (20-200 microg kg(-1)) polynomial regression of ITX showed a relative standard deviation (sdv) of +/-1.51 % (r = 0.99981). Starting with the limit of quantification the response was linear (sdv = +/-2.18 %, r = 0.99893). Regarding repeatability (n = 9) a coefficient of variation (CV) of 1.1 % was obtained for ITX at 32 ng on silica gel plates and of 2.9 % on reversed-phase plates. Repeatabilities (n = 4) of ITX determination at 20, 50 and 100 microg kg( 1) in milk, yoghurt, soybean oil and margarine showed CVs between +/-1.0 and 6.4 %. The results prove that modern planar chromatography is a rapid and cost efficient alternative method to quantify ITX in milk-based or fatty matrices. Only positive results are confirmed by online ESI/MS in the SIM mode (LOQ 128 pg) and by DART/MS involving a minimal employment of the MS device, which is a further advantage of HPTLC. Overall mean recovery rates of ITX at 20 or 50 and 100 microg kg(-1) (n = 8) were 41 % for milk, 70 % for yoghurt, 6 % for margarine and 12 % for soy bean oil. However, with the internal standard correction recoveries were about 130 % for milk and yoghurt and 70 and 97 % for margarine and soy bean oil, respectively. PMID- 16715279 TI - Integration of microcolumns and microfluidic fractionators on multitasking centrifugal microfluidic platforms for the analysis of biomolecules. AB - This work demonstrates the development of microfluidic compact discs (CDs) for protein purification and fractionation integrating a series of microfluidic features, such as microreservoirs, microchannels, and microfluidic fractionators. The CDs were fabricated with polydimethylsiloxane (PDMS), and each device contained multiple identical microfluidic patterns. Each pattern employed a microfluidic fractionation feature with operation that was based on the redirection of fluid into an isolation chamber as a result of an overflow. This feature offers the advantage of automated operation without the need for any external manipulation, which is independent of the size and the charge of the fractionated molecules. The performance of the microfluidic fractionator was evaluated by its integration into a protein purification microfluidic architecture. The microfluidic architecture employed a microchamber that accommodated a monolithic microcolumn, the fractionator, and an isolation chamber, which was also utilized for the optical detection of the purified protein. The monolithic microcolumn was polymerized "in situ" on the CD from a monolith precursor solution by microwave-initiated polymerization. This technique enabled the fast, efficient, and simultaneous polymerization of monoliths on disposable CD microfluidic platforms. The design of the CD employed allows the integration of various processes on a single microfluidic device, including protein purification, fractionation, isolation, and detection. PMID- 16715280 TI - Different iron-chelating properties of pyochelin diastereoisomers revealed by LC/MS. AB - Pyochelin is a siderophore and virulence factor common to Burkholderia cepacia and several Pseudomonas strains. It is isolated from bacterial media as a mixture of two epimers, which readily equilibrate in most solvents. Experiments based on high-performance liquid chromatography/electrospray ionization mass spectrometry are reported here, allowing the investigation of the different Fe(III)-chelating properties of pyochelin diastereomers in solution without the need for labourious isolation. It is demonstrated in this study that only one of the two pyochelin diastereomers is able to chelate Fe(III); no Fe(III) complexes of the other diastereomer could be detected. The Fe(III)-pyochelin complex exhibited a 1:1 metal-to-siderophore ratio and no evidence for other stoichiometries was found. PMID- 16715281 TI - Development of a new standard reference material: SRM 1955 (homocysteine and folate in human serum). AB - Total homocysteine (tHCY) and folate are interrelated biomarkers for arteriosclerosis and coronary heart disease. Although many different methods for both tHCY and folate are clinically available, the intermethod and interlaboratory results are often poor, resulting in the need for a matrix reference material and reference methods. The National Institute of Standards and Technology (NIST) has developed isotope dilution liquid chromatography/mass spectrometry (LC/MS) and liquid chromatography/ tandem mass spectrometry (LC/MS/MS) methods for determination of tHCY and several folate forms including 5 methyltetrahydrofolic acid (5MT) and folic acid (FA). Additionally, a method for simultaneous measurement of tHCY, 5MT, and FA has been developed and validated. In collaboration with the Centers for Disease Control and Prevention (CDC), mass spectrometric methods and methods used in clinical laboratories have been applied to characterize a new Standard Reference Material (SRM), SRM 1955, "Homocysteine and Folate in Human Serum," containing low, medium, and high levels of tHCY and 5MT. Additionally, FA, 5-formyltetrahydrofolic acid (5FT), vitamin B12, and total folate values are provided. Use of the new SRM should improve clinical measurements and will permit traceability to internationally recognized certified reference materials, as described by European Directive 98/79/EC on in vitro diagnostic medical devices. PMID- 16715283 TI - Headspace solid-phase microextraction gas chromatography tandem mass spectrometry for the determination of brominated flame retardants in environmental solid samples. AB - A headspace solid-phase microextraction gas chromatography coupled with tandem mass spectrometry (HSSPME-GC-MS-MS) methodology for determination of brominated flame retardants in sediment and soil samples is presented. To the best of our knowledge, this is the first time that SPME has been applied to analyze polybrominated biphenyls (PBBs) and polybrominated diphenyl ethers (PBDEs) in environmental solid samples. Analyses were performed using 0.5-g solid samples moisturized with 2 mL water, employing a polydimethylsiloxane (PDMS) fiber coating, exposed to the headspace at 100 degrees C for 60 min. Several types of environmental solid samples were included in this study and the extraction efficiency was related to the organic matter content of the sample. Calibration was performed using real samples, and the method showed good linearity over a wide concentration range, precision, and afforded quantitative recoveries. The obtained detection limits were in the sub-ng g(-1) for all the target analytes in both samples. The proposed procedure was applied to several marine and river sediments and soils, some of which were found to contain PBDEs at concentrations in the ng g(-1) level; BDE-47, BDE-100, and BDE-99 were the major congeners detected. The proposed method constitutes a rapid and low-cost alternative for the analysis of the target brominated flame retardants in environmental solid samples, since the clean-up steps, fractionation, and preconcentration of extracts inherent to the classical multi-step solvent extraction procedures are avoided. PMID- 16715282 TI - Trace analysis and occurrence of anhydroerythromycin and tylosin in influent and effluent wastewater by liquid chromatography combined with electrospray tandem mass spectrometry. AB - Two wastewater treatment plants (WWTPs) of northern Colorado were monitored for anhydroerythromycin and tylosin. An analytical method has been developed and validated for the trace determination and confirmation of these compounds in the raw influent and final effluent water matrices. This method was used to evaluate the occurrence and fate of these compounds in WWTPs. The method uses solid-phase extraction and liquid chromatography-tandem mass spectrometry with positive electrospray ionization. Detection and quantification was performed using selected reaction monitoring, and a method detection limit of between 0.01 and 0.06 microg/L was obtained. Unequivocal confirmation analysis of analyte identity according to the criteria (based on the use of identification points) of the 2002/657/EC European Commission Decision was possible with satisfactory results. Average recoveries for the two compounds ranged from 89.2+/-9.7% for raw influent to 93.7+/-6.9% for effluent wastewaters. The within-run precision of the assay was found to be always less than 14.1% for the two analytes. The overall precision was always less than 13.7%. The relative uncertainty of the present assay was also evaluated and the combined relative uncertainty ranged from 6.4 to 15.5% over three days of the validation study. These compounds were partially removed in the WWTPs with a removal efficiency of >50%. The measured concentrations in raw influents and effluents ranged from 0.09-0.35 and 0.04-0.12 microg/L for anhydroerythromycin to 0.06-0.18 and ND-0.06 microg/L for tylosin, respectively. The results indicate that WWTP effluents are relevant point sources for residues of these compounds in the aquatic environment. These occurrence results were compared with those in WWTP wastewaters of other countries. PMID- 16715284 TI - Mineral oil content in sediments and soils: comparability, traceability and a certified reference material for quality assurance. AB - The performance of twelve laboratories with previously established proficiency in the determination of the mineral oil content in a fresh water sediment is described. The summation parameter total petrol hydrocarbon (TPH) is defined according to ISO 16703:2004 with regard to the sample preparation to be applied, the flame ionisation detection (FID) and the boiling range of C10-C40 to be integrated. Comprehensive tests of homogeneity and stability have been carried out on the candidate material using appropriate models. The outcome of the study served as the basis for the certification of the candidate reference material as ERM-CC015a. The certified mass fraction is 1,820+/-130 mg kg(-1) and traceability was established by using an appropriate calibration standard certified for the mass fraction of C10-C40. The interlaboratory scatter of measurement results in this exercise can largely be explained by the variability of the individual calibrations based on this common calibration standard. PMID- 16715285 TI - Detection of toxic effects of Cd2+ on different fish species via liver cytochrome P450-dependent monooxygenase activities and FTIR spectroscopy. AB - The in vivo and in vitro effects of Cd2+ and the CYP1A inductor beta naphthoflavone(beta-NF) on the hepatic cytochrome P450 (Cyt 450) monooxygenases were studied in silver carp (Hypophthalmichtys molitrix V.), wels (Silurus glanis L.), and carp (Cyprinus carpio). In vivo treatment of carp with a high dose of Cd2+ (10 mg kg(-1), for 3 days) caused a strong inhibition of 7-ethoxyresorufin-O deethylase (EROD) and a lower inhibition of 7-ethoxycoumarin-O-deethylase (ECOD) activity. The low-dose cadmium treatment (2 mg kg(-1) Cd2+, for 6+3 days) resulted in 4-fold increase in EROD and a 3-fold increase in ECOD activity. The combined treatment with Cd2+ and beta-NF in both cases led to a loss of EROD inducibility. The silver carp and wels were treated with 10 mg L(-1) Cd2+ for 72 h in water. The Cyt P450 content in the wels liver microsomes was increased significantly after treatment for 48 h, whereas there was only a slight, not significant increase in Cyt P450 content in the silver carp microsomes. While the Cd2+ treatment resulted in inhibition of the CYP1A isoenzymes (EROD and ECOD), the APND (aminopyrene-N-demethylase, CYP2B or CYP3A isoenzyme) activity was increased 3- to 4-fold in both fish species. In vitro experiments of the effect of Cd2+ led to a concentration-dependent inhibition in all three investigated fish species. The ECOD isoenzyme of silver carp was the most sensitive to Cd2+. The lowest concentration of Cd2+ resulted in 50% inhibition. The APND isoenzyme was similarly sensitive to Cd2+ in all three investigated fish species. The most sensitive species was the wels, and the least sensitive were the carp isoenzyme. FTIR spectroscopy confirmed that cadmium caused damage to the protein structure. These results support the enzyme activity measurements measured in vivo and in vitro. PMID- 16715287 TI - First trimester embryonic/fetal heart rate in normal pregnant women. AB - OBJECTIVE: To establish reference ranges for first trimester embryonic/fetal heart rate in normal pregnant women. MATERIALS AND METHODS: This was a cross sectional study. We performed ultrasonogram in 319 normal pregnant women, gestation age between 6(+0) and 14(+6) weeks and measured embryonic/fetal heart rates using M-mode. The embryonic/fetal heart rates were analyzed according to gestational ages (GA). RESULTS: Data of 319 pregnancies were analyzed and the outcome revealed the mean of fetal heart rate according to gestational age. The mean embryonic heart rates during the 6th-7th week of gestation are lower when compared with the other gestational ages. At the 8th week of gestation, the embryonic heart rate is maximum. Then the embryonic/fetal heart rate gradually decreased to 161 beats/min at the 14th week of gestation. The regression equation for embryonic/fetal heart rate (beat/min) [y] according to GA (day) [x] was y = 53.124 + 6.1333x + (-0.0407)x ( 2 )(r (2) = 0.525; P < 0.001). CONCLUSION: Embryonic/fetal heart rates during the 6(+0) and 14(+6) week of gestation are related to GA. Our reference ranges may be useful for further studies such as for prediction of adverse pregnancy outcome in threatened abortion. PMID- 16715288 TI - Assessing the reproductive performance of men co-infected with HIV-1 and hepatitis C undergoing assisted reproduction. AB - OBJECTIVE: To assess the reproductive performance of men co-infected with hepatitis C virus (HCV) and human immunodeficiency virus (HIV-1) undergoing assisted reproduction. DESIGN: A retrospective analysis. SETTING: University based assisted reproductive technology (ART) center. PATIENTS: We reviewed 217 consecutive ART cycles performed on 106 HIV-1 serodiscordant couples between August 1997 and March 2004, in which 28 men (26%) were seropositive for HCV and HIV-1 (group 1). MAIN OUTCOME MEASURES: Patient demographics and laboratory testing were compared to reproductive outcomes of men undergoing similar treatment but infected only with HIV-1 (group 2). RESULTS: Co-infected men and their partners were of similar age as men infected only with HIV-1. Comparing group 1 to group 2, like values were noted for HIV-1 viral loads [1,993 +/- 1,140 copies/ml (mean +/- SE) vs. 1,659 +/- 487 copies/ml]; CD4 counts (520 +/- 98 vs. 604 +/- 38 mm(-3)); and semen parameters. IVF performance and outcomes were similar, with fertilization rate (0.68 +/- 0.03 vs. 0.71 +/- 0.02); number of normally cleaving embryos (6.0 +/- 0.5 vs. 5.3 +/- 0.3); embryo implantation rate (0.27 +/- 0.04 vs. 0.2 +/- 0.02); and clinical pregnancy rate (40 vs. 29%). Although the male mortality rate was low in both groups, morbidity among co infected men was significantly higher. Seven of 28 men (25%) had detectable HCV viral loads, and 14 (50%) had elevated liver function tests. CONCLUSION: Men co infected with HCV and HIV-1 do reasonably well undergoing ART to prevent transmission of viruses to their partners and children. PMID- 16715289 TI - Assessment of uterine cavity by hysteroscopy in assisted reproduction programme and its influence on pregnancy outcome. AB - OBJECTIVE: The aim of the study was to evaluate if the diagnosis and treatment of uterine cavity abnormalities by hysteroscopy in patients undergoing IVF programme is of any value in improving clinical pregnancy outcome. METHODS: 520 patients participated in this prospective randomized study and were classified into two groups. Group I (n = 265) without office hysteroscopy. Group II (n = 255) had office hysteroscopy and was sub classified into Group II a and Group II b. Group II a (n = 160) had normal hysteroscopic findings whereas Group II b (n = 95) had abnormal office hysteroscopy findings, which were corrected at the same time. RESULT: There was no difference in the mean number of oocytes retrieved, fertilization rate, and number of embryos transferred among the patients in different groups. Statistically significant difference was observed in terms of clinical pregnancy rates between Group I and Group II a (26.2 and 44.44%, P < 0.05), and Group I and Group II b (26.2 and 39.55%, P < 0.05), respectively. CONCLUSION: Patients with recurrent IVF embryo transfer failures after normal hysterosalpingography findings should also be reevaluated using hysteroscopy prior to further commencing IVF-embryo transfer cycles in order to enhance the clinical pregnancy rates. PMID- 16715290 TI - Increased peri- and post-elective cesarean section morbidity in women infected with human immunodeficiency virus-1: a case-controlled multicenter study. AB - OBJECTIVE: Although elective cesarean section (ECS) is the currently recommended modality for delivering women infected with the human immunodeficiency virus (HIV), historical evidence suggests that they are at higher risk of postoperative complications than noninfected women. Those risks have to be carefully balanced against the presumed minimal benefit of ECS, especially in the case of low viral load and high CD4 counts. We therefore compared the incidence and type of post ECS complications in HIV-infected women, most with low viral loads and high CD4 cell counts, with those in matched noninfected women treated by the same surgical teams. STUDY DESIGN: A Swiss 8-center, prospective, matched case-control study compared minor and major post-ECS complication prevalence, hospital stay and confounding factors (surgeon experience) between HIV-infected and noninfected women. RESULTS: Minor complications in the 53 matched pairs were eightfold more frequent overall in infected women. More frequent specific minor complications were anemia, blood loss and urinary tract infection. Yet the surgeons performing ECS in infected women were more experienced. Complications prolonged hospital stay in infected women. Major complication rates did not significantly differ between the groups. CONCLUSION: HIV-positive women have a higher risk of post-ECS morbidity, even with high CD4 counts and low viral load. Therefore, the blanket recommendation of ECS in HIV-infected women requires a review. PMID- 16715291 TI - The effect of bilateral oophorectomy on bone mineral density. AB - The objective of this study is to investigate the effect of bilateral oophorectomy with total abdominal hysterectomy on bone loss, comparing the cases having surgery before and after the menopause. Bone mineral density (BMD) measurements were obtained from the lumbar spine and femoral neck of totally 127 cases. Out of 127, 105 had surgery before menopause and 22 cases were operated on postmenopausally. The results were compared with the USA normal values. The average age of surgical menopause (SM) cases was 48.45 years with a mean duration of menopause of 5.77 years. The average height and weight were 157.67 cm and 68.19 kg, respectively. The average age of cases having surgery after menopause (SAM) was 62.45 years with a mean duration of 5.59 years after the surgery (duration after menopause is 13.23 years). The average height and weight were 157.45 cm and 73.55 kg, respectively. The average of BMD measurements of lumbar spines L2-L4 was 1.04 gr/cm(2) (BMD = 85.65% and T score = -0.96) in the cases with SM. On the contrary, the average of the BMD measurements of lumbar spines L2 L4 was 1.05 gr/cm(2) (BMD = 101.14% and T score = 0.24) in the cases with SAM. The average of the BMD measurements of femoral neck was 0.85 gr/cm(2) (BMD = 91.39% and T score = -0.64) in the cases with SM. On the contrary, the average of the BMD measurements of femoral neck was 0.82 gr/cm(2) (BMD = 96.69% and T score = -0.31) in the cases with SAM. The bilateral oophorectomy as a surgical procedure is not a statistically significant factor for the acceleration of the bone loss. The main points are the age and the duration of menopause of the patient affecting the bone loss if the surgery is performed before menopause. PMID- 16715292 TI - The association between total antioxidant status and oxidative stress in Behcet's disease. AB - There has recently been growing evidence supporting the importance of oxidative stress in the pathogenesis of Behcet's disease (BD). In this study, we aim to evaluate total antioxidant status (TAS) and total oxidative stress (TOS) in BD patients, and compare their results both with controls that had recurrent aphthous stomatitis (RAS) and healthy controls. TAS statistically decreased in RAS patients, and TOS levels increased in BD and RAS patients than those in healthy controls. The serum levels of Cu significantly increased only in BD patients when compared with healthy controls. Fe levels were not statistically different among the BD patients, RAS patients and healthy controls, but there was a positive correlation between TOS and plasma Fe levels in BD patients. Our results suggest that there is an insufficient antioxidant system and increased oxidative status both in BD and RAS patients. The antioxidant supplementations in addition to medical treatments will improve the quality of life. PMID- 16715293 TI - The enhancement of HCN channel instantaneous current facilitated by slow deactivation is regulated by intracellular chloride concentration. AB - The hyperpolarization-activated cation current I (f) plays a key role in the modulation of rhythmic activity in cardiac pacemaker cells and spontaneously firing neurons. I (f) is generated by hyperpolarization-activated cyclic nucleotide-gated channels (HCN1-HCN4) and comprises two components: the fast instantaneous current (I (INS)) and the slowly developing steady-state current (I (SS)). We found that in I (f) traces evoked by consecutive hyperpolarization, the I (INS) amplitude of the second trace was up to 50% larger than the first. I (SS) was identical. This pre-hyperpolarization mediated enhancement of I (INS) was maximal in channels displaying slow kinetics (sinoatrial I (f), HCN3, and HCN4), while it was almost negligible for fast channels (HCN1 and HCN2). The enhancement quantitatively correlated with the frequency of hyperpolarization. Analysis of HCN4 currents suggested that enhancement was facilitated by incomplete deactivation, confirmed by HCN2-HCN4 chimeric studies. It is important to note that intracellular Cl(-) was found to be a cellular suppressor of I (INS) enhancement. Cl(-) inhibited the enhancement with an IC(50) around 25 mM and Hill coefficients between 2 and 6. Cl(-) shifted V (0.5) by +7 mV when [Cl(-)](i) was increased from 11 to 141 mM. In conclusion, I (INS) represents a quantitatively important component of I (f) at low Cl(-) (as found in most cell types). Moreover, an increase in cellular Cl(-) will suppress enhancement of I (INS) and, hence, potentially affect the electrical properties of cells. PMID- 16715294 TI - An alpha3beta4 subunit combination acts as a major functional nicotinic acetylcholine receptor in male rat pelvic ganglion neurons. AB - We identified major subunits of the nicotinic acetylcholine receptor (nAChR) involved in excitatory postsynaptic potential and intracellular Ca(2+) ([Ca(2+)]i) increase in the major pelvic ganglion (MPG) neurons of the male rat. ACh elicited fast inward currents in both sympathetic and parasympathetic MPG neurons. Mecamylamine, a selective antagonist for alpha3beta4 nAChR, potently inhibited the ACh-induced currents in sympathetic and parasympathetic neurons (IC(50); 0.53 and 0.22 microM, respectively). Furthermore, alpha-conotoxin AuIB (10 microM), a new selective antagonist for alpha3beta4 nAChR, blocked more than 80% of the ACh-induced currents in MPG neurons. Conversely, alpha-bungarotoxin, alpha-methyllycaconitine, and dihydro-beta-erythroidine, known as blockers of the alpha7 or alpha4beta2, did not show selective blocking effects on MPG neurons. ACh transiently increased [Ca(2+)]i which was subsequently abolished in the extracellular Ca(2+)-free environment. Simultaneous recording of [Ca(2+)]i and ionic currents revealed that ACh increased [Ca(2+)]i under the conditions of the voltage-clamped (at -80 mV) state, and this resulted from the influx through nAChR itself. ACh-induced [Ca(2+)]i increase was blocked by mecamylamine (10 microM), but was not affected by atropine (1 microM). RT-PCR analysis showed that, among subunits of nAChR, alpha3 and beta4 were predominantly expressed in MPG. We suggest that activation of alpha3 and beta4 nAChR subunits in MPG neurons induce fast inward currents and [Ca(2+)]i increase, possibly mediating a major role in pelvic autonomic synaptic transmission. PMID- 16715295 TI - Successful implantation of intravenously administered stem cells correlates with severity of inflammation in murine myocarditis. AB - The present study was designed to determine whether cardiac inflammation is important for the successful homing of stem cells to the heart after intravenous injection in a murine myocarditis model. Male Bagg albino/c mice were infected with encephalomyocarditis virus (EMCV) to produce myocarditis. Subgroups of mice received single injections by tail vein of embryonic stem cells (ESCs) transfected with green fluorescent protein (GFP) as a marker at days 3, 14, or 60 after infection; other subgroups without stem cell injections were killed at each of these time points to assess the degree of inflammation present. The surviving mice were killed at day 90 after virus infection and hemodynamics, gross pathology, histology, and inflammatory cytokine production in the hearts were measured. Our results indicate that myocardial inflammation was most severe and cytokine production highest at day 14 after EMCV inoculation, and in particular, was strongly positive for interleukin 6. Mice receiving intravenous ESC injections on day 14 after EMCV inoculation showed the largest number of GFP positive cells at the time of death and the greatest functional improvement compared to uninfected controls without inflammation. We conclude that factors released from myocardium during inflammation are important for enhancing the homing, migration, and implantation of systemically infused stem cells. PMID- 16715296 TI - Chloride and bicarbonate have similar affinities to the intestinal anion exchanger DRA (down regulated in adenoma). AB - DRA (down regulated in adenoma, SLC26A3) is an anion exchanger that mediates electroneutral NaCl absorption in the ileum and proximal colon together with NHE3 (Na/H exchanger isoform 3), and that is involved in duodenal and possibly pancreatic bicarbonate secretion. Thus, its chloride and bicarbonate affinities are important for both processes. [Cl]i and pHi transients were measured using MQAE and BCECF. HEK293 cells stably expressing DRA were exposed to 0 mM Cl at various [HCO3] (9 to 51 mM, at 5% CO2 or 15 to 57 mM, at pH 7.5) to determine the HCO3 affinity. After intracellular Cl depletion, 10, 30, and 90 mM Cl were readded at various [HCO3]s to determine the relative Cl and HCO3 affinities. The k0.5 for extracellular HCO3 is between 18.5 and 32.8 mM. Cl and HCO3 compete with similar affinities for transport by DRA. DRA activity is independent of pHo between 7.0 and 7.75. DRA is activated by alkaline pHi. Competition of Cl and HCO3 does not significantly impair NaCl absorption, because in the ileum and colon, luminal Cl is comparably high. Activation at alkaline pHi supports functional coupling of DRA and NHE3 by the subapical pHi. In the distal pancreatic ductal system, luminal HCO3 is high compared to luminal Cl. Under these conditions, competition of Cl and HCO3 is difficult to reconcile with a role of DRA in Cl reabsorption in exchange for HCO3. Our data, thus, provide indirect evidence against a role of DRA in pancreatic HCO3 secretion. PMID- 16715297 TI - [Abdominal compartment syndrome]. AB - Abdominal compartment syndrome (ACS) is characterized by a persistent pathologic increase in intra-abdominal pressure (IAP) exceeding 20 mmHg with consecutive dysfunction of multiple organ systems. The main causes of ACS are abdominal trauma, obstruction, infection, and sepsis, but it may also be initiated by extra abdominal diseases. The gold standard for diagnosis is repeated assessment of the IAP measurements of bladder pressure. The incidence of ACS is up to 15% in operative ICUs and the therapy of choice for it is decompressive laparotomy. Nevertheless, mortality is high, up to 60%. PMID- 16715299 TI - Comparison of five prognostic scoring systems, the French-American-British (FAB) and World Health Organization (WHO) classifications in patients with myelodysplastic syndromes: Results of a single-center analysis. AB - We retrospectively studied 89 consecutive patients diagnosed with primary myelodysplastic syndrome (MDS) over a period of 10 years to (1) identify prognostic factors for overall survival (OS) and leukemia-free survival (LFS); (2) to assess and compare the Bournemouth-, Spanish-, Dusseldorf-, Lille-, and the International prognostic scoring systems (IPSS); and to (3) compare the French-American-British (FAB) and World Health Organization (WHO) classifications. The median age of patients was 63 years (range, 26-85). Karyotype analyses were done in 85 patients (96%). Median OS was 3 years; 67 patients (75%) have died, and 28 (31%) had progression to acute myeloid leukemia (AML). Major independent prognostic variables for both OS and LFS (multivariate analysis) were percentage of bone marrow (BM) blasts (P < 0.0001), and in patients with cytogenetic data available, cytogenetic risk groups by Lille-score (OS, P = 0.031/LFS, P = 0.002) and IPSS (OS, P = 0.024). All five prognostic scoring systems successfully discriminated risk groups as regards OS and LFS, but in patients with cytogenetic data available, the major independent prognostic score for OS (P < 0.0001) and LFS (P = 0.006) was the IPSS. The FAB and WHO classifications also successfully discriminated between risk groups. The new WHO subgroups [refractory cytopenia with multilineage dysplasia (RCMD), with (RCMD RS) or without ringed sideroblasts] showed a significantly (P = 0.0454) different prognosis for OS, but not for LFS (P = 0.0839), in comparison to the subgroups having erythroid dysplasia only (RA/RARS). Risk stratification into refractory anemia with excess blast-I (RAEB-I) and RAEB-II tended to yield different prognoses for OS and LFS. The 5q-minus syndrome strongly predicted for a good prognosis. In patients treated with the demethylating agent decitabine (n = 24), IPSS "poor risk" cytogenetics were unable to predict for the expected worse prognosis when compared to "intermediate-risk" cytogenetics. In conclusion, we confirm in a single-center patient cohort that the use of the WHO classification improves the predictive value of the FAB classification and that, in patients with cytogenetic data available, the IPSS can be used for clinical decision making. PMID- 16715303 TI - Experiments on the Fehrer-Raab effect and the 'Weather Station Model' of visual backward masking. AB - The Fehrer-Raab effect (simple reaction time is unaffected by metacontrast masking of the test stimulus) seems to imply that a stimulus can trigger a voluntary reaction without reaching a conscious representation. However, it is also possible that the mask triggers the reaction, and that the masked test stimulus causes a focussing of attention from which processing of the mask profits, thus reaching conscious representation earlier. This is predicted by the Weather Station Model of visual masking. Three experiments tested this explanation. Experiment 1 showed that the masked test stimulus caused a temporal shift of the mask. Experiment 2 showed that the reaction in the Fehrer-Raab effect was not exclusively triggered by a conscious representation of the test stimulus: the mask was involved in evoking the reaction. Experiment 3 again revealed a temporal shift of the mask. However, the shift was only about half as large as the Fehrer-Raab effect. The psychometric functions suggested that the observers used two different cues for their temporal order judgments. The results cast doubts on whether judged temporal order yields a direct estimate of the time of conscious perception. Some methodological alternatives are discussed. PMID- 16715302 TI - Long-term results and complications following uvulopalatopharyngoplasty in 116 consecutive patients. AB - A modified uvulopalatopharyngoplasty (UPPP) was carried out between January 1992 and December 2003 at the ENT Department of the Inselspital in Bern in 146 patients with habitual or complicated rhonchopathy. The operation consisted of a classical tonsillectomy or residual tonsil resection and additional shortening of the uvula. The natural mucosal fold between the uvula and the upper pole of the tonsils was carefully preserved. A wide opening to the rhinopharynx was created by asymmetric suturing of the glossopalantine and pharyngopalatine arches. A retrospective questionnaire with regard to rhonchopathy, phases of apnea, daytime drowsiness, obstruction of nasal breathing, long-term complications and patient satisfaction was used to evaluate the short-term and long-term effectiveness of the modified UPPP as well as the incidence of adverse side effects. Complete postoperative courses were evaluated in 116 patients. Surgical complications were restricted to one case with postoperative hemorrhage. A velum insufficiency or postoperative rhinopharyngeal stenosis did not occur. Eighty-three patients (72%) confirmed a persistent suppression or substantial improvement of the rhonchopathy. Disappearance or decrease of sleep apnea was confirmed in 12 (63%) out of 19 postoperative polysomnographic follow-up investigations. Long-term complications occurred in a total of 27 (23%) of 116 patients. They were confined to minor problems such as dryness of the mouth (n = 12), slight difficulty in swallowing (n = 7), discrete speech disturbances (n = 1), and slight pharyngeal dysesthesias (n = 7) with feeling of a lump in the throat and compulsive clearing of the throat. Eighty-five patients (73%) reported that they were satisfied with the postoperative result even several years after the operation. Looking back, 31 patients (27%) would no longer have the operation performed. The inadequate result of the rhonchopathy was specified as the reason by 21 patients. Ten patients had unpleasant memories of the operation because of intensive postoperative pain. Snoring and apneic phases are suppressed or improved by non traumatic UPPP in the majority of patients. This effect persisted even years after the operation. PMID- 16715305 TI - Comment on the article: a study of clinical predictors of lumbar provocation discography (M. Laslett et al.). PMID- 16715304 TI - Complement insufficiency limits efficacy in a xenograft model of hyperacute rejection for cancer therapy. AB - Hyperacute rejection (HAR) is a rapid immunological response to an organ xenotransplant caused by recognition of endothelial galactose(alpha1,3)galactose (alphaGal) epitopes and complement-mediated cell lysis by host anti-alphaGal antibody ('natural antibody'). The alphaGal epitope is synthesised by a galactosyl transferase ((alpha1,3)GT) which humans lack. Because human cells transduced with (alpha1,3)GT are sensitised to natural antibody/complement mediated lysis in human serum, delivery of (alpha1,3)GT to tumour vasculature in patients is a potential therapeutic strategy, by mimicking the pathophysiology of organ rejection. We therefore sought to develop an animal model of HAR for cancer therapy. Nude/(alpha1,3)GT knock-out mice allowed the growth of human tumour xenografts and the use of ecotropic retrovirus producer cells to generate expression of alphaGal on tumour vasculature. Lysis of alphaGal-positive murine endothelial cells with rabbit complement in conjunction with murine anti-alphaGal antibody in vitro was used to define the conditions necessary for HAR. However, tumour growth retardation and destruction of alphaGal-positive tumour endothelium were minimal after their administration, despite sera retaining post hoc cytolytic activity with fresh complement. The major limitation of this experimental system, of relevance to other therapeutic approaches, results from the use of a xenograft, in which additional xenoreactivities lead to complement insufficiency. Development of a tractable preclinical model in which to evaluate HAR for cancer therapy requires a syngeneic experimental system. PMID- 16715306 TI - Preadolescent presentation of a lumbar chordoma: results of vertebrectomy and fibula strut graft reconstruction at 8 years. AB - Chordoma is a tumour of notochordal origin which usually involves the sacrum or skull base presenting in adulthood. Chordoma in a mobile spinal segment is infrequent and the authors report an extremely rare presentation of L3 chordoma in a child aged 7 years. Although a benign tumour, mobile segment chordoma is more locally aggressive, more likely to metastasise and has a poorer 5 year survival than sacral and clival lesions. Wide surgical excision and reconstruction is the treatment of choice in vertebral chordoma. This case was treated with staged vertebrectomy and fibular strut graft reconstruction and the results of clinical and radiological follow up at 8 years are presented. PMID- 16715307 TI - Internet based multicenter study for thoracolumbar injuries: a new concept and preliminary results. AB - This article reports about the internet based, second multicenter study (MCS II) of the spine study group (AG WS) of the German trauma association (DGU). It represents a continuation of the first study conducted between the years 1994 and 1996 (MCS I). For the purpose of one common, centralised data capture methodology, a newly developed internet-based data collection system ( http://www.memdoc.org ) of the Institute for Evaluative Research in Orthopaedic Surgery of the University of Bern was used. The aim of this first publication on the MCS II was to describe in detail the new method of data collection and the structure of the developed data base system, via internet. The goal of the study was the assessment of the current state of treatment for fresh traumatic injuries of the thoracolumbar spine in the German speaking part of Europe. For that reason, we intended to collect large number of cases and representative, valid information about the radiographic, clinical and subjective treatment outcomes. Thanks to the new study design of MCS II, not only the common surgical treatment concepts, but also the new and constantly broadening spectrum of spine surgery, i.e. vertebro-/kyphoplasty, computer assisted surgery and navigation, minimal invasive, and endoscopic techniques, documented and evaluated. We present a first statistical overview and preliminary analysis of 18 centers from Germany and Austria that participated in MCS II. A real time data capture at source was made possible by the constant availability of the data collection system via internet access. Following the principle of an application service provider, software, questionnaires and validation routines are located on a central server, which is accessed from the periphery (hospitals) by means of standard Internet browsers. By that, costly and time consuming software installation and maintenance of local data repositories are avoided and, more importantly, cumbersome migration of data into one integrated database becomes obsolete. Finally, this set-up also replaces traditional systems wherein paper questionnaires were mailed to the central study office and entered by hand whereby incomplete or incorrect forms always represent a resource consuming problem and source of error. With the new study concept and the expanded inclusion criteria of MCS II 1, 251 case histories with admission and surgical data were collected. This remarkable number of interventions documented during 24 months represents an increase of 183% compared to the previously conducted MCS I. The concept and technical feasibility of the MEMdoc data collection system was proven, as the participants of the MCS II succeeded in collecting data ever published on the largest series of patients with spinal injuries treated within a 2 year period. PMID- 16715308 TI - Movement coordination of the lumbar spine and hip during a picking up activity in low back pain subjects. AB - The effect of low back pain, with or without nerve root signs, on the joint coordination and kinematics of the lumbar spine and hips during everyday activities, such as picking up an object from the floor, are largely unknown. An experimental study was designed to compare lumbar spine and hip joint kinematics and coordination in subjects with and without sub-acute low back pain, while picking up an object in a sitting position. A three-dimensional real-time electromagnetic tracking device was used to measure movements of the lumbar spine and hips. Sixty participants with subacute low back pain, with or without straight leg raise signs, and twenty healthy asymptomatic participants were recruited. The ranges of motions of lumbar spine and hips were determined. Movement coordination between the two regions was examined by cross-correlation. Results showed that mobility was significantly reduced in subjects with back pain, who compensated for limited motion through various strategies. The contribution of the lumbar spine relative to that of the hip was, however, found to be similar in all groups. The lumbar spine-hip joint coordination was substantially altered in subjects with back pain, in particular, those with a positive straight leg raise sign. We conclude that changes in the lumbar and hip kinematics were related to back pain and limitation in straight leg raise. Lumbar hip coordination was mainly affected by the presence of positive straight leg raise sign when picking up an object in a sitting position. PMID- 16715312 TI - Large-scale mutation screening in patients with dilated or hypertrophic cardiomyopathy: a pilot study using DGGE. AB - Cardiomyopathies are complex myocardial diseases characterized by inappropriate ventricular hypertrophy (HCM) or dilation (DCM). Both disorders may lead to sudden death or progressive heart failure and exhibit familial aggregation with marked genetic heterogeneity. Many candidate genes were identified by linkage analysis, experimental animal studies, and expression analysis. A systematic assessment of the prevalence of different mutations in these genes requires high throughput analyses. In this paper, we present a simple and reliable protocol for mutation screening by heteroduplex analysis which reduced costs and workload of sequencing. Employing denaturing gradient gel electrophoresis (DGGE), 11 known and 14 potential candidate genes for HCM and DCM were analyzed. DGGE assays allowed analysis of 286 of the 312 protein coding exons, performing only four alternative polymerase chain reaction protocols and only two different DGGE analysis conditions. Sensitivity for the detection of heteroduplexes proved excellent, even for GC-rich DNA fragments, which were analyzed by a combination of DGGE and constant denaturant gel electrophoresis. To confirm DGGE sensitivity in cases where no variants in our human DNA samples could be observed, we generated heteroduplexes from homologous human and chimpanzee DNA. The platform proved a valuable contribution to elucidating the genetic causes of DCM and HCM as demonstrated by the identification of 17 different known and novel mutations and 98 different polymorphisms in our setting. PMID- 16715313 TI - The treatment of intrahepatic calculosis by applying helix hydro-jet lithotripsy under video choledochoscope: a report of 30 cases. AB - BACKGROUND AND AIMS: Intrahepatic lithiasis is a common disease in southeast Asia [Sheen-Chen and Chou, Acta Chir Scand 156:387-390, 1990], and a difficult problem of biliary surgery. There is no established method of treating patients with intrahepatic stones [Uchiyama et al., Arch Surg 137:149-533, 2002]. In recent years, resection of the affected liver lobe or segment is the best therapeutic option to completely remove the source of recurrent infection. The need for endoscopic treatment modalities is evident because hepatic resections are combined with a high morbidity and mortality rate [Andersson et al., HPB Surg 2:145-147, 1990; Adamek et al., Scand J Gastroenterol 34:1157-1161, 1999]. Hepatic resection only fit the cases in which the stones localized in one lobe or segment, while it doesn't fit the cases which have polystones in left and right biliary tract. Duodenoscope can only get the stones in the common bile duct and cannot deal with the intrahepatic lithiasis. The management of intrahepatic lithiasis can only be treated by intraoperative or postoperative choledochoscope. For big stones or compact stones, lithotripsy should be applied. But the laser lithotripsy and the electrohydraulic lithotripsy can cause serious complications such as perforation of bile duct. It needs a safer and more reliable treatment for intrahepatic lithiasis. The aim of our work is to study the lithotrity treatment of intrahepatic lithiasis by using helix hydro-jet under Video Choledochoscope. MATERIALS AND METHODS: From March 31, 2003 to October 20, 2004, 30 intrahepatic stone patients were treated. Eighteen of them were women and 12 were men, with ages ranging from 35 to 80 years (mean, 58 years). According to B ultrasound and computed tomography (CT) scan report, there were five cases of intrahepatic lithiasis and common bile duct stones, 25 cases of left and right hepatic duct stones, and one case with giant intrahepatic stone (1.5 x 1.5 x 1.2 cm). Intraoperative or postoperative choledochoscopic helix hydro-jet lithotripsy was applied through a video choledochoscope. For the patients to have the intraoperative choledochoscopic helix hydro-jet lithotripsy, they should be diagnosed correctly by B ultrasound or CT scan. The biliary tract reconstruction by spiral CT scan is as helpful as MRCP or ERCP for clinical diagnosis. For the patients to have the postoperative choledochoscopic helix hydro-jet lithotripsy, they should be diagnosed correctly by T-tube cholangiography and BUS and CT scan. All patients should be verified without stones remaining in the bile duct after lithotripsy by choledochoscopic examination and T-tube cholangiography, and should be examined by BUS again after 6 months to 1 year. We decide whether complications occurred by observation of symptoms and signs after choledochoscopy and lithotripsy. RESULTS: Seventy-five intrahepatic stones with diameter ranging from 0.6 to 1.5 cm were successfully fragmentized in 30 patients using of helix hydro-jet lithotripsy. These fragmentized stones mainly are bilirubin stones. The lithotripsy was carried for 45 times and the procedure needs 1-1.5 h. Helix hydro jet lithotripsy are used in 16 cases during operation and 12 cases after operation; two cases during operation and after operation. Intrahepatic calculosis was cleaned out completely and verified by postoperative choledochoscope examination and postoperative T-tube cholangiography examination. No complications were observed. CONCLUSION: Helix hydro-jet lithotripsy under video choledochoscope is a safe and effective method for the removal of intrahepatic stone. No bile duct damnified and perforation was observed. The procedure is without pain and heat, and the pressure can be adjusted easily. The research provides a new way of using the helix hydro-jet, and a new way of curing the intrahepatic lithiasis. PMID- 16715316 TI - Avulsion of anterior cruciate ligament from femoral condyle: an unusual case report and a review of the literature. AB - We report a rare case of avulsion of anterior cruciate ligament from the lateral femoral condyle and describe the arthroscopic management of such a case along with a review of the literature. PMID- 16715315 TI - Dynamic patchplasty--a tension-free reconstruction of incisional hernias. AB - BACKGROUND AND AIMS: An anatomical and physiological reconstruction of the abdominal wall after incisional hernia must include reconstruction of the linea alba to restore the physiological function of the vertebral column and respiration. Against this background, a special reconstruction method employing a mesh, which is performed in our department, is presented. MATERIALS AND METHODS: In the period between 1 January 2000 and 1 June 2004, a total of 106 patients with incisional hernias after median laparotomy of the upper and lower abdomen were operated on in our department using this tension-free technique with one or two meshes. We have now evaluated the outcomes after a median follow-up of 25.8 months (6.5-53.3). RESULTS: In one patient (0.9%), the peritoneal sac was inadvertently opened, resulting in injury to the small bowel serosa. Surgical complications were observed in 14 patients (13.2%) after dynamic patchplasty. In seven cases (6.6%), wound healing disorders were seen. Overall, 6 out of 106 (5.5%) patients needed revision surgery. In the follow-up period, 92 patients (follow-up 86.8%) could be contacted and re-examined. Four patients (3.8%) had died. There were four recurrences (4.3%), which had already been repaired, and one chronic seroma, which caused no symptoms and which was detected in ultrasound and computed tomography. CONCLUSION: In view of the possibility of achieving physiological reconstruction and in the light of a tolerable morbidity rate, dynamic patchplasty is a suitable method for incisional hernias. Long-term results hold out promise of an acceptable low recurrence rate. PMID- 16715317 TI - Modulation of depth-dependent properties in tissue-engineered cartilage with a semi-permeable membrane and perfusion: a continuum model of matrix metabolism and transport. AB - The functional properties of cartilaginous tissues are determined predominantly by the content, distribution, and organization of proteoglycan and collagen in the extracellular matrix. Extracellular matrix accumulates in tissue-engineered cartilage constructs by metabolism and transport of matrix molecules, processes that are modulated by physical and chemical factors. Constructs incubated under free-swelling conditions with freely permeable or highly permeable membranes exhibit symmetric surface regions of soft tissue. The variation in tissue properties with depth from the surfaces suggests the hypothesis that the transport processes mediated by the boundary conditions govern the distribution of proteoglycan in such constructs. A continuum model (DiMicco and Sah in Transport Porus Med 50:57-73, 2003) was extended to test the effects of membrane permeability and perfusion on proteoglycan accumulation in tissue- engineered cartilage. The concentrations of soluble, bound, and degraded proteoglycan were analyzed as functions of time, space, and non-dimensional parameters for several experimental configurations. The results of the model suggest that the boundary condition at the membrane surface and the rate of perfusion, described by non dimensional parameters, are important determinants of the pattern of proteoglycan accumulation. With perfusion, the proteoglycan profile is skewed, and decreases or increases in magnitude depending on the level of flow-based stimulation. Utilization of a semi-permeable membrane with or without unidirectional flow may lead to tissues with depth-increasing proteoglycan content, resembling native articular cartilage. PMID- 16715318 TI - The effect of nucleus pulposus crosslinking and glycosaminoglycan degradation on disc mechanical function. AB - Altered mechanical loading, secondary to biochemical changes in the nucleus pulposus, is a potential mechanism in disc degeneration. An understanding of the role of this altered mechanical loading is only possible by separating the mechanical and biological effects of early nucleus pulposus changes. The objective of this study was to quantify the mechanical effect of decreased glycosaminoglycans (GAG) and increased crosslinking in the nucleus pulposus using in vitro rat lumbar discs. Following initial mechanical testing the discs were injected according to the four treatment groups: PBS control, chondroitinase-ABC (ChABC) for GAG degradation, genipin (Gen) for crosslinking, or a combination of chondroitinase and genipin (ChABC+Gen). After treatment the discs were again mechanically tested, followed by histology or biochemistry. Neutral zone mechanical properties were changed by approximately 20% for PBS, ChABC, and ChABC+Gen treatments (significant only for PBS in a paired comparison). These trends were reversed with genipin crosslinking alone. With ChABC treatment the effective compressive modulus increased and the GAG content decreased; with the combination of ChABC+Gen the mechanics and GAG content were unchanged. Degradation of nucleus pulposus GAG alters disc axial mechanics, potentially contributing to the degenerative cascade. Crosslinking is unlikely to contribute to degeneration, but may be a potential avenue of treatment. PMID- 16715320 TI - Microstructural modeling of collagen network mechanics and interactions with the proteoglycan gel in articular cartilage. AB - Cartilage matrix mechanical function is largely determined by interactions between the collagen fibrillar network and the proteoglycan gel. Although the molecular physics of these matrix constituents have been characterized and modern imaging methods are capable of localized measurement of molecular densities and orientation distributions, theoretical tools for using this information for prediction of cartilage mechanical behavior are lacking. We introduce a means to model collagen network contributions to cartilage mechanics based upon accessible microstructural information (fibril density and orientation distributions) and which self-consistently follows changes in microstructural geometry with matrix deformations. The interplay between the molecular physics of the collagen network and the proteoglycan gel is scaled up to determine matrix material properties, with features such as collagen fibril pre-stress in free-swelling cartilage emerging naturally and without introduction of ad hoc parameters. Methods are developed for theoretical treatment of the collagen network as a continuum-like distribution of fibrils, such that mechanical analysis of the network may be simplified by consideration of the spherical harmonic components of functions of the fibril orientation, strain, and stress distributions. Expressions for the collagen network contributions to matrix stress and stiffness tensors are derived, illustrating that only spherical harmonic components of orders 0 and 2 contribute to the stress, while orders 0, 2, and 4 contribute to the stiffness. Depth- and compression-dependent equilibrium mechanical properties of cartilage matrix are modeled, and advantages of the approach are illustrated by exploration of orientation and strain distributions of collagen fibrils in compressed cartilage. Results highlight collagen-proteoglycan interactions, especially for very small physiological strains where experimental data are relatively sparse. These methods for determining matrix mechanical properties from measurable quantities at the microscale (composition, structure, and molecular physics) may be useful for investigating cartilage structure-function relationships relevant to load-bearing, injury, and repair. PMID- 16715319 TI - Analysis of the relationship between peak stress and proteoglycan loss following injurious compression of human post-mortem knee and ankle cartilage. AB - While traumatic joint injuries are known to increase the risk of osteoarthritis (OA), the mechanism is not known. Models for injurious compression of cartilage may identify predictors of injury that suggest a clinical mechanism. We investigated the relationship between peak stress during compression and glycosaminoglycan (GAG) loss after injury for knee and ankle cartilages. Human cartilage explant disks were harvested post-mortem from the knee and ankle of three organ donors with no history of OA and subjected to injurious compression to 65% strain in uniaxial unconfined compression at 2 mm/s (400%/s). The GAG content of the conditioned medium was measured 3 days after injury. After injury of knee cartilage disks, damage was visible in 18 of 39 disks (36%). Three days after injury, the increase in GAG loss to the medium (GAG loss from injured disks minus GAG loss from location-matched uncompressed controls) was 1.5+/-0.3 microg/disk (mean +/- SEM). With final strain and compression velocity held constant, we observed that increasing peak stress during injury was associated with less GAG loss after injury (P<0.001). In contrast, ankle cartilage appeared damaged after injury in only 1 of 16 disks (6%), there was no increase in GAG loss (0.0+/-0.3 microg/disk), and no relationship between peak stress and increase in GAG loss was detected (P=0.51). By itself, increasing peak stress did not appear to be an important cause of GAG loss from human cartilage in our injurious compression model. However, we observed further evidence for differences in the response of knee and ankle cartilages to injury. PMID- 16715321 TI - Total hip replacement in patients with severe bleeding disorders. A 30 years single center experience. AB - Arthropathy of the hip is moderate in frequency in haemophiliac patients, but is less common than ankle, knee or elbow arthropathy. We report about our experience with total hip replacement in patients with severe bleeding disorders over a period of 30 years. Between July 1972 and 2002, 15 hips in 13 patients were replaced. The main bleeding disorders were Haemophilia A in ten patients and severe v. Willebrand disease in three patients. The mean follow-up was 132 months (range 12-363). We can demonstrate good long-term results, with only one aseptic loosening after 14 years and one septic loosening after 14 months in an HIV positive patient. The Harris Hip Score increased from 48 points (32-66) preoperatively to 89 (76-100) postoperatively. In conclusion, total hip replacement performed in a specialised haemophiliac centre is a safe procedure, and results in pain relief and improvement of the quality of life in patients with severe bleeding disorders. PMID- 16715322 TI - Immunohistochemical localization and quantitative assessment of GnRH-, FSH-, and LH-receptor mRNA Expression in canine skin: a powerful tool to study the pathogenesis of side effects after spaying. AB - It has been proposed that gonadotropins and/or gonadotropin releasing hormone (GnRH) could be involved in the pathophysiology of the side effects after spaying in bitches, such as urinary incontinence and an increased production of a woolly undercoat. In order to provide tools to investigate the role of these hormones in dogs we developed immunohistochemical techniques and real-time RT-PCR to study whether GnRH-, LH-, and FSH-receptors exist in canine skin and urinary bladder. Tissue samples from the skin of the flank region and the ventral midline of the urinary bladder from euthanised dogs were examined. We were able to quantify mRNA expression of GnRH-, FSH-, and LH-receptors in canine skin and bladder biopsies with a high primer efficacy. Immunohistochemical studies showed that GnRH-, FSH-, and LH-receptors are expressed in vessel walls, the epidermis, the hair follicle and in sebaceous and sweat glands in canine skin and in transitional epithelium, and smooth muscle tissue in the urinary bladder. Our data provide the fundamentals to examine the distribution of FSH-, LH-, and GnRH-receptors in canine skin and urinary bladder and to assess gene activity at the transcriptional level by real-time RT-PCR. PMID- 16715323 TI - A retrospective mortality study of workers exposed to radon in a Brazilian underground coal mine. AB - Recently a high radon concentration was detected in the underground coal mine of Figueira, located in the south of Brazil. This coal mine has been operating since 1942 without taking cognizance of the high radon environment. In order to assess possible radon-related health effects on the workers, a retrospective (1979-2002) mortality study of 2,856 Brazilian coal miners was conducted, with 2,024 underground workers potentially exposed to radon daughters. Standard mortality ratio (SMR) analysis hints at lower mortality from all causes for both underground (SMR = 88, 95% CI = 78-98) and surface workers (SMR = 96, 95% CI = 80 114). A high statistically significant SMR for lung cancer mortality was observed only in the underground miners (SMR = 173, 95% CI = 102-292), with a statistically significant trend reflecting the duration of underground work. High statistically significant SMRs were observed for pneumonia as a cause of death between both surface (SMR = 304, 95% CI = 126-730) and underground miners (SMR = 253, 95% CI = 140-457). Because mortality from smoking-related cancers other than lung cancer was not found elevated in underground workers and because diesel equipments were not used in this mine, it can be concluded that the enhanced lung cancer mortality observed for underground miners is associated with exposure to radon and radon daughters, rather than other confounding risk factors. PMID- 16715325 TI - Amplitude-integrated EEG (aEEG) predicts outcome after cardiac arrest and induced hypothermia. AB - OBJECTIVE: To evaluate the use of continuous amplitude-integrated EEG (aEEG) as a prognostic tool for survival and neurological outcome in cardiac arrest patients treated with hypothermia. DESIGN: Prospective, observational study. SETTING: Multidisciplinary intensive care unit in a university hospital. INTERVENTION: Comatose survivors of cardiac arrest were treated with induced hypothermia for 24 h. An aEEG recording was initiated upon arrival at the ICU and continued until the patient regained consciousness or, if the patient remained in coma, no longer than 120 h. The aEEG recording was not available to the ICU physician, and the aEEG tracings were interpreted by a neurophysiologist with no knowledge of the patient's clinical status. Only clinically visible seizures were treated. MEASUREMENTS AND RESULTS: Thirty-four consecutive hypothermia-treated cardiac arrest survivors were included. At normothermia (mean 37 h after cardiac arrest), the aEEG pattern was discriminative for outcome. All 20 patients with a continuous aEEG at this time regained consciousness, whereas 14 patients with pathological aEEG patterns (flat, suppression-burst or status epilepticus) did not regain consciousness and died in hospital. Patients were evaluated neurologically upon discharge from the ICU and after 6 months, using the Cerebral Performance Category (CPC) scale. Eighteen patients were alive with a good cerebral outcome (CPC 1--2) at 6-month follow-up. CONCLUSION: A continuous aEEG pattern at the time of normothermia was discriminative for regaining consciousness. aEEG is an easily applied method in the ICU setting. PMID- 16715324 TI - Diagnostic strategy in cancer patients with acute respiratory failure. AB - OBJECTIVE: Nearly 15% of cancer patients experience acute respiratory failure (ARF) requiring admission to the intensive care unit, where their mortality is about 50%. This review focuses on ARF in cancer patients. The most recent literature is reviewed, and emphasis is placed on current controversies, most notably the risk/benefit ratio of fiberoptic bronchoscopy and BAL in patients with severe hypoxemia. BACKGROUND: Fiberoptic bronchoscopy with bronchoalveolar lavage (BAL) is the cornerstone of the causal diagnosis. However, the low diagnostic yield of about 50%, related to the widespread use of broad-spectrum antimicrobial therapy in cancer patients, has generated interest in high resolution computed tomography (HRCT) and primary surgical lung biopsy. In patients with hypoxemia, bronchoscopy and BAL may trigger a need for invasive mechanical ventilation, thus considerably decreasing the chances of survival. DISCUSSION: The place for recently developed, effective, noninvasive diagnostic tools (tests on sputum, blood, urine, and nasopharyngeal aspirates) needs to be determined. The prognosis is not markedly influenced by cancer characteristics; it is determined chiefly by the cause of ARF, need for mechanical ventilation, and presence of other organ failures. Although noninvasive ventilation reduces the need for endotracheal intubation and diminishes mortality rate, its prolonged use in patients with severe disease may preclude optimal diagnostic and therapeutic management. The appropriateness of switching to endotracheal mechanical ventilation in patients who fail noninvasive ventilation warrants evaluation. CONCLUSION: This review discusses risks and benefits from invasive and non invasive diagnostic and therapeutic strategies in critically ill cancer patients with acute respiratory failure. Avenues for research are also suggested in order to improve survival in these very high risk patients. PMID- 16715327 TI - Influence of severity of illness, medication and selective decontamination on defecation. AB - OBJECTIVE: To describe the pattern of defecation in critically ill ventilated patients and the influence of selective decontamination (SDD) and other medication. DESIGN: Descriptive cohort study. SETTING: Mixed surgical-medical ICU in a university Hospital. PATIENTS: Ventilated patients with a length of stay >or=7 days taking part in a study on SDD. MEASUREMENTS: Daily registration of defecation, SOFA (sepsis-related organ failure assessment score) score, administration of dopamine, noradrenaline, morphine and other medications. RESULTS: The first defecation occurred after a mean of 6.2 days. Patients with defecation within 6[Symbol: see text]days had lower mean SOFA scores, received more cisapride and lactulose and less dopamine, noradrenaline and morphine, and had a shorter duration of mechanical ventilation and ICU stay. On 57% of the days, no stools were produced; on 31% diarrhea, and on 12%, normal stools. Patients receiving SDD had more days with normal stools and less with diarrhea. Diarrhea was preceded by the administration of lactulose in the majority of patients. CONCLUSION: Time to first defecation correlated with severity of illness, vasoactive medication, administration of morphine, cisapride and lactulose, duration of mechanical ventilation and length of stay. Diarrhea seemed at least partially iatrogenic. PMID- 16715326 TI - Constant flow insufflation of oxygen as the sole mode of ventilation during out of-hospital cardiac arrest. AB - BACKGROUND: Constant flow insufflation of oxygen (CFIO) through a Boussignac multichannel endotracheal tube has been reported to be an efficient ventilatory method during chest massage for cardiac arrest. METHODS: Patients resuscitated for out-of-hospital cardiac arrest were randomly assigned to standard endotracheal intubation and mechanical ventilation (MV; n =457) or use of CFIO at a flow rate of 15 l/min (n=487). Continuous chest compressions were similar in the two groups. Pulse oximetry level was recorded every 5[Symbol: see text]min. Outcome of initial resuscitation, hospital admission, complications, and discharge from the intensive care unit (ICU) were analyzed. The randomization scheme was changed during the study, but the in-depth analysis was performed only on the first cohort of 341 patients with CFIO and 355 with MV, because of randomization problems in the second part. RESULTS: No difference in outcome was noted regarding return to spontaneous circulation (CFIO 21%, MV 20%), hospital admission (CFIO 17%, MV 16%), or ICU discharge (CFIO 2.4%, MV 2.3%). The level of detectable pulse saturation and the proportion of patients with saturation above 70% were higher with CFIO. Ten patients with MV but only one with CFIO had rib fractures. CONCLUSIONS: CFIO is a simplified alternative to MV, with favorable effects regarding oxygenation and fewer complications, as observed in this group of patients with desperate prognosis. PMID- 16715328 TI - Procalcitonin kinetics in pediatric patients with systemic inflammatory response after open heart surgery. AB - OBJECTIVE: To evaluate procalcitonin and C-reactive protein as markers of inflammation severity and their value in predicting development of organ failure after pediatric open heart surgery. DESIGN: Prospective, observational, clinical study. SETTING: Single university hospital. PATIENTS: Thirty-three pediatric patients with systemic inflammatory response syndrome (SIRS; n=19) and SIRS+organ failure (SIRS+OF; n=14) following open heart surgery were included. MEASUREMENTS AND RESULTS: Plasma procalcitonin and C-reactive protein levels were measured before and after the operation, and 1, 2, 3, and 4 days after surgery. Patients were evaluated daily to assess organ failure. Postoperative procalcitonin levels in the SIRS+OF group were significantly higher than in the SIRS group. C-reactive protein levels were similar between the groups throughout the study period. Peak procalcitonin levels were found to be positively correlated with aortic cross clamp and cardiopulmonary bypass times, duration of mechanical ventilation, intensive care unit and hospital stay, mortality and organ failure development. Peak procalcitonin was found to be a good predictor of postoperative organ failure development and mortality. However, the predictive value of peak C reactive protein for organ failure and mortality was found to be weak. Double peak procalcitonin curves were observed in SIRS+OF patients with infection during the intensive care unit stay. CONCLUSION: In the SIRS+OF group peak procalcitonin levels were found to be highly predictive for mortality and organ failure development, whereas C-reactive protein levels were not. Daily procalcitonin measurements in SIRS+OF patients may help identify the postoperative infection during the follow-up period. PMID- 16715329 TI - Weaning failure from cardiovascular origin. PMID- 16715342 TI - Remembrance: celebrating the man behind the science. PMID- 16715343 TI - Desire for children and pregnancy risk behavior among HIV-infected men and women in Uganda. AB - To identify ways to improve prevention of mother-to-child transmission (PMTCT) of HIV, we conducted a cross-sectional study of 1,092 HIV-infected men and women attending an AIDS support organization in Jinja, Uganda, between October 2003 and June 2004. Pregnancy risk behavior was defined as having sex without contraceptive or condom. Overall, 42% of participants were sexually active, 33% practiced pregnancy risk behavior, and 18% desired more children. Men were almost four times to want more children than the women (27% vs. 7%). Among those practicing pregnancy risk behavior, 73% did not want more children and were at high risk for unwanted pregnancies. Although 81% knew that mother-to-child transmission of HIV could be prevented, only 22% believed that an HIV-infected woman who received PMTCT therapy could still deliver an HIV-infected child. Lack of MTCT information, having attended the program for 95% of treatments. CONCLUSIONS: The present study identified and validated a dosing nomogram and schedule to deliver low and nontoxic suramin concentrations that produce chemosensitization in preclinical models. PMID- 16715362 TI - Effect of oropharyngeal length in drug lung delivery via suspension pressurized metered dose inhalers. AB - PURPOSE: To determine the effect of the oropharyngeal length in adults on the lung dose of a suspension pressurized metered dose inhaler, and whether employing small volume spacers can alter this role. METHODS: Depositions of Ventolin Evohaler (100) microg in the oropharyngeal models of two healthy adult subjects with 17.1 cm (short cast) and 19.9 cm (long cast) centerline lengths via three small volume spacers [two spacers with 3 cm effective length but one with 6.5 cm2 (L3) and the other with 24.6 cm2 (L3W) cross sections, and the Optimiser] were studied. RESULTS: Without using spacers, lung dose of the long cast (19.52 +/- 2.32 microg, mean +/- standard deviation) was significantly larger than that for the short cast (8.08 +/- 1.01 microg, p < 0.006). However, using the L3 spacer with the short cast made the lung dose (18.59 +/- 3.33 microg) similar to that for the long cast alone. Lung doses of the short cast (20.43 +/- 1.42 microg) and the long cast (30.81 +/- 1.84 microg) with the L3W spacer were similar to those with the L3 spacer. However, using the Optimiser spacer increased the lung dose for the short cast (22.27 +/- 6.03 microg) and significantly for the long cast (35.61 +/- 2.19 microg, p < 0.006) compared to those for the L3 spacer. Using spacers increased drug deposition in the oropharynx part of the short cast, and this reduced the lung dose compared to that for the long cast. CONCLUSION: The oropharyngeal length in adults may affect the lung dose via the pMDIs, which may not be eliminated by using small volume spacers. PMID- 16715363 TI - Oral absorption of poorly water-soluble drugs: computer simulation of fraction absorbed in humans from a miniscale dissolution test. AB - PURPOSE: The purpose of this study was to develop a new system for computer simulation to predict fraction absorbed (F(a)) of Biopharmaceutical Classification System (BCS) class II (low solubility-high permeability) drugs after oral administration to humans, from a miniscale dissolution test. METHODS: Human oral absorption of 12 lipophilic drugs was simulated theoretically by using the dissolution and permeation parameters of the drugs. A miniscale dissolution test and a solubility study were carried out in a conventional buffer and a biorelevant medium (pH 6.5). A dissolution parameter, which can simulate in vivo dissolution, was obtained from the in vitro dissolution curve. Human intestinal permeability was estimated assuming that the permeation was limited by diffusion through the unstirred water layer. The F(a) in humans was predicted and then compared with clinical data. RESULTS: The dissolution and solubility of most model drugs were faster and higher in a biorelevant medium than in a conventional buffer. The simulated absorption was limited by the drug dissolution rate and/or solubility. Predicted F(a) was significantly correlated with clinical data (correlation coefficient r2 = 0.82, p < 0.001) when the dissolution profiles in biorelevant medium were used for the simulation. CONCLUSIONS: This new system quantitatively simulated human absorption and would be beneficial for the prediction of human F(a) values for BCS class II drugs. PMID- 16715364 TI - Reverse hexagonal phase nanodispersion of monoolein and oleic acid for topical delivery of peptides: in vitro and in vivo skin penetration of cyclosporin A. AB - PURPOSE: To obtain and characterize reverse hexagonal phase nanodispersions of monoolein and oleic acid, and to evaluate the ability of such system to improve the skin penetration of a model peptide (cyclosporin A, CysA) without causing skin irritation. METHODS: The nanodispersion was prepared by mixing monoolein, oleic acid, poloxamer, and water. CysA was added to the lipid mixture to obtain a final concentration of 0.6% (w/w). The nanodispersion was characterized; the skin penetration of CysA was assessed in vitro (using porcine ear skin mounted in a Franz diffusion cell) and in vivo (using hairless mice). RESULTS: The obtainment of the hexagonal phase nanodispersion was demonstrated by polarized light microscopy, cryo-TEM and small angle X-ray diffraction. Particle diameter was 181.77 +/- 1.08 nm. At 0.6%, CysA did not change the liquid crystalline structure of the particles. The nanodispersion promoted the skin penetration of CysA both in vitro and in vivo. In vitro, the maximal concentrations (after 12 h) of CysA obtained in the stratum corneum (SC) and in the epidermis without stratum corneum (E) + dermis (D) were approximately 2 fold higher when CysA was incorporated in the nanodispersion than when it was incorporated in the control formulation (olive oil). In vivo, 1.5- and 2.8-times higher concentrations were achieved in the SC and [E+D], respectively, when the nanodispersion was employed. No histopathological alterations were observed in the skin of animals treated with the nanodispersion. CONCLUSION: These results demonstrate that the hexagonal phase nanodispersion is effective in improving the topical delivery of peptides without causing skin irritation. PMID- 16715361 TI - Plasmid engineering for controlled and sustained gene expression for nonviral gene therapy. AB - Gene therapy requires the introduction of genetic material in diseased cells with the aim of treating or ultimately curing a disease. Since the start of gene therapy clinical trials in 1990, gene therapy has proven to be possible, but studies to date have highlighted the difficulty of achieving efficient, specific, and long-term transgene expression. Efforts to improve gene therapy strategies over the past years were mainly aimed at solving the problem of delivery, without paying much attention to the optimization of the expression cassette. With the current understanding of the eukaryotic transcription machinery and advanced molecular biology techniques at our disposition, it has now become possible to create custom-made transgene expression cassettes optimized for gene therapy applications. In this review, we will discuss several strategies that have been explored to improve the level and duration of transgene expression, to increase control over expression, or to restrict transgene expression to specific cell types or tissues. Although still in its infancy, such strategies will eventually lead to improvement of nonviral gene therapy and expansion of the range of possible therapeutic applications. PMID- 16715366 TI - Glucose partition coefficient and diffusivity in the lower skin layers. AB - PURPOSE: This work aims to estimate the diffusivity and partitioning of glucose in the dermis and the viable epidermis of human skin. METHODS: The partition coefficient of glucose between phosphate-buffered saline and dermis, tape stripped epidermis (TSE), stratum corneum (SC), and split-thickness skin, was measured in vitro using human cadaver skin. Glucose permeability across dermis and tape-stripped split-thickness skin (TSS) was measured using side-by-side diffusion cells. Glucose desorption from TSE and human epidermal membrane (HEM) was measured. All measurements were conducted at 32 degrees C. RESULTS: The partition coefficient for glucose [mean +/- SD (no. of samples)] was 0.65 +/- 0.09 (n = 25) for dermis, 0.81 +/- 0.06 (n = 10) for TSE, and 0.53 +/- 0.12 (n = 9) for SC. Glucose diffusivity in dermis was calculated to be 2.64 +/- 0.42 x 10( 6) cm2/s (n = 14). Glucose diffusivities in the viable epidermis estimated from TSS permeation, TSE desorption, and HEM desorption were 0.075 +/- 0.050 x 10(-6) cm2/s (n = 5), 0.037 +/- 0.018 x 10(-6) cm2/s (n = 4), and 1.0 +/- 0.6 x 10(-6) cm2/s (n = 4), respectively. CONCLUSION: The tissue/buffer partition coefficient of glucose in all skin layers was found to be less than unity, suggestive of excluded volumes in each layer. Glucose diffusivity in human dermis was found to be one third of its value in water, indicative of hindered diffusion related to the structural components of the tissue. A substantially lower value for glucose diffusivity in viable epidermis is suggested. PMID- 16715365 TI - FTIR and nDSC as analytical tools for high-concentration protein formulations. AB - PURPOSE: The aim of the study is to evaluate Fourier-transform infrared spectroscopy (FTIR) as an analytical tool for high-concentrated protein formulations. METHODS: FTIR is used to determine the melting temperature (T(m (FTIR))) of various proteins, such as bovine serum albumin (BSA), immunoglobulin (IgG1), beta-lactoglobulin (beta-LG), and lysozyme (HEWL), at different protein concentrations (5-100 mg/mL), where four data interpretation methods are discussed. The obtained T(m (FTIR)) values are further compared to the T(m) measured by the nanodifferential scanning calorimetry (nDSC) technique. RESULTS: The T(m (FTIR)) values of IgG1 and beta-LG showed strong consistency and corresponded to the nDSC results irrespective of the method of data interpretation and the protein concentration applied. In contrast, the T(m (FTIR)) of BSA and HEWL is characterized by significant deviations. Only the midpoint of the second-derivative intensity-temperature curve of the intermolecular beta-sheet mode measured at a concentration of 100 mg/mL is consistent with the nDSC results. CONCLUSIONS: Determination of a T(m (FTIR)) is feasible by the midpoint of the intensity-temperature plot of the arising intermolecular beta-sheet band. More significant results are obtained for proteins, which are predominantly composed of intramolecular beta-sheet elements as well as at higher protein concentrations. A further study was started to assess the predictability of long-term protein stability by T(m (FTIR)). PMID- 16715367 TI - Hepatic CYP3A expression is attenuated in obese mice fed a high-fat diet. AB - PURPOSE: Changes in physiological, pathophysiological, and/or nutritional conditions often alter the expression of drug-metabolizing enzymes. In this study, we investigated obesity-induced changes in hepatic cytochrome P450 (P450) levels using nutritionally obese mice. METHODS: To induce obesity, mice were fed a high-fat diet or treated with gold thioglucose, which impairs ventromedial hypothalamus. Total RNAs and microsomal and nuclear proteins were prepared from the liver of these mice, and mRNA and protein levels of P450s and transcription factors were determined. RESULTS: Among P450s examined, the constitutive expression of CYP3As was drastically reduced at both mRNA and protein levels by nutrition-induced obesity. One-week administration of a high-fat diet also reduced hepatic CYP3As. However, changes in nuclear receptors involved in the transcriptional regulation of CYP3A genes were not correlated with that of CYP3As. Obese mice induced by gold thioglucose exhibited a different expression profile of hepatic P450s with no significant change in CYP3As. CONCLUSION: High fat diet-induced changes in energy metabolism, which eventually result in obesity, modulate the hepatic expression profile of P450s, particularly CYP3As. Alternatively, the accumulation of a certain component in a high-fat diet may directly attenuate the CYP3A expression, suggesting a clinically important drug diet interaction. PMID- 16715369 TI - Complement activation by core-shell poly(isobutylcyanoacrylate)-polysaccharide nanoparticles: influences of surface morphology, length, and type of polysaccharide. AB - PURPOSE: Biodistribution of intravenously administered nanoparticles depends on opsonization. The aim of this study was the evaluation of complement activation induced by nanoparticles coated with different polysaccharides. Influences of size and configuration of dextran, dextran sulfate, or chitosan bound onto nanoparticles were investigated. METHOD: Core-shell nanoparticles were prepared by redox radical or anionic polymerization of isobutylcyanoacrylate in the presence of polysaccharides. Conversion of C3 into C3b in serum incubated with nanoparticles was evaluated. RESULTS: Cleavage of C3 increased with size of dextran bound in "loops" configuration, whereas it decreased when dextran was bound in "brush." It was explained by an increasing steric repulsive effect of the brush, inducing poor accessibility to OH groups. The same trend was observed for chitosan-coated nanoparticles. Nanoparticles coated with a brush of chitosan activated the complement system lesser than nanoparticles coated with a brush of dextran. This was explained by an improved repelling effect. Dextran-sulfate coated nanoparticles induced a low cleavage of C3 whereas it strongly enhanced protein adsorption. CONCLUSION: Complement activation was highly sensitive to surface features of the nanoparticles. Type of polysaccharide, configuration on the surface, and accessibility to reactive functions along chains are critical parameters for complement activation. PMID- 16715368 TI - Effects of antipsychotic drugs on I(to), I (Na), I (sus), I (K1), and hERG: QT prolongation, structure activity relationship, and network analysis. AB - PURPOSE: To evaluate in vitro and computationally model the effects of selected antipsychotic drugs on several ionic currents that contribute to changes in the action potential in cardiac tissue. METHODS: Fourteen antipsychotic drugs or metabolites were examined to determine whether QT interval prolongation could be accounted for by an effect on one or more myocardial ion channels [I(to), I(Na), I(sus), I(K1), and human ether-a-go-go related gene (hERG)]. Using the patch clamp technique, drug effects on these human cardiac currents were tested. RESULTS: All molecules had little inhibitory effect on ion channels (blocking at concentrations >5 microM) other than hERG. A significant correlation was observed between the estimated hERG blockade and the increase in corrected QT for five of the antipsychotics. Molecular modeling identified hydrophobic features related to the interaction with hERG and correctly rank-ordered the test set molecules olanzapine and its metabolites. A network analysis of ligand and protein interactions around hERG using MetaCore (GeneGo Inc., St. Joseph, MI, USA) was used to visualize antipsychotics with affinity for this channel and their interactions with other proteins in this database. CONCLUSION: The antipsychotics do not inhibit the ion channels I(to), I(Na), I(sus), I(K1) to any appreciable extent; however, blockade of hERG is a likely mechanism for the prolongation of the QT interval. PMID- 16715370 TI - ATP-binding cassette transporter G2 mediates the efflux of phototoxins on the luminal membrane of retinal capillary endothelial cells. AB - PURPOSE: The purpose of this study was to clarify the localization and function of the ATP-binding cassette transporter G2 (ABCG2; BCRP/MXR/ABCP) in retinal capillary endothelial cells, which form the inner blood-retinal barrier, as an efflux transport system. METHODS: The expression was determined by reverse transcriptase polymerase chain reaction and Western blotting. The localization was identified by immunostaining. The transport function of ABCG2 was measured by flow cytometry. RESULTS: Western blotting indicated that ABCG2 was expressed as a glycosylated disulfide-linked complex in the mouse retina and in peripheral tissues, including liver, kidney, and small intestine. Double immunolabeling of ABCG2 and glucose transporter 1 suggested that ABCG2 was localized on the luminal membrane of mouse retinal capillary endothelial cells. ABCG2 mRNA and protein were found to be expressed in a conditionally immortalized rat retinal capillary endothelial cell line, TR-iBRB, and rat retina. Treatment with Ko143, an ABCG2 inhibitor, restored the accumulation of pheophorbide a and protoporphyrin IX in TR-iBRB cells. CONCLUSION: ABCG2 is expressed on the luminal membrane of retinal capillary endothelial cells, where ABCG2 acts as the efflux transporter for photosensitive toxins such as pheophorbide a and protoporphyrin IX. ABCG2 could play an important role at the inner blood-retinal barrier in restricting the distribution of phototoxins and xenobiotics in retinal tissue. PMID- 16715371 TI - Metabonomics techniques and applications to pharmaceutical research & development. AB - In this review, the background to the approach known as metabonomics is provided, giving a brief historical perspective and summarizing the analytical and statistical techniques used. Some of the major applications of metabonomics relevant to pharmaceutical Research & Development are then reviewed including the study of various influences on metabolism, such as diet, lifestyle, and other environmental factors. The applications of metabonomics in drug safety studies are explained with special reference to the aims and achievements of the Consortium for Metabonomic Toxicology. Next, the role that metabonomics might have in disease diagnosis and therapy monitoring is provided with some examples, and the concept of pharmacometabonomics as a way of predicting an individual's response to treatment is highlighted. Some discussion is given on the strengths and weaknesses, opportunities of, and threats to metabonomics. PMID- 16715372 TI - Enhanced oral paclitaxel bioavailability after administration of paclitaxel loaded lipid nanocapsules. AB - PURPOSE: The aim of this study was to evaluate the pharmacokinetics of paclitaxel loaded lipid nanocapsules (LNC) in rats to assess the intrinsic effect of the dosage form on the improvement of paclitaxel oral exposure. METHODS: Paclitaxel loaded LNC were prepared and characterized in terms of size distribution, drug payload, and the kinetics of paclitaxel crystallization. Taxol, Taxol with verapamil, or paclitaxel-loaded LNC were administered orally to rats. The plasma concentration of paclitaxel was determined using liquid chromatography mass spectrometry. RESULTS: The average size of LNC was 60.9 +/- 1.5 nm. The drug payload of paclitaxel was 1.91 +/- 0.01 mg/g of aqueous dispersion. The encapsulation efficiency was 99.9 +/- 1.0%, and 1.7 +/- 0.1% of paclitaxel was crystallized after 24 h. The oral bioavailability of Taxol alone was 6.5%. After oral administration of paclitaxel-loaded LNC or paclitaxel associated with verapamil, the area under the plasma concentration-time curve was significantly increased (about 3-fold) in comparison to the control group (p < 0.05). CONCLUSIONS: The results indicated that LNC provided a promising new formulation to enhance the oral bioavailability of paclitaxel while avoiding the use of pharmacologically active P-gp inhibitors, such as verapamil. PMID- 16715373 TI - Adaptive neuro-fuzzy modeling of poorly soluble drug formulations. AB - PURPOSE: The purpose of this study was to evaluate the efficiency of a neuro fuzzy logic-based methodology to model poorly soluble drug formulations and predict the development of the particle size that has been proven to be an important factor for long-term stability. METHODS: An adaptive neuro-fuzzy inference system was used to model the natural structures within the data and construct a set of fuzzy rules that can subsequently used as a predictive tool. The model was implemented in Matlab 6.5 and trained using 75% of an experimental data set. Subsequently, the model was evaluated and tested using the remaining 25%, and the predicted values of the particle size were compared to the ones from the experimental data. The produced adaptive neuro-fuzzy inference system-based model consisted of four inputs, i.e., acetone, propylene glycol, POE-5 phytosterol (BPS-5), and hydroxypropylmethylcellulose 90SH-50, with four membership functions each. Moreover, 256 fuzzy rules were employed in the model structure. RESULTS: Model training resulted in a root mean square error of 4.5 x 10(-3), whereas model testing proved its highly predictive efficiency, achieving a correlation coefficient of 0.99 between the actual and the predicted values of the particle size (mean diameter). CONCLUSIONS: Neuro-fuzzy modeling has been proven to be a realistic and promising tool for predicting the particle size of drug formulations with an easy and fast way, after proper training and testing. PMID- 16715374 TI - Protein structural conformation and not second virial coefficient relates to long term irreversible aggregation of a monoclonal antibody and ovalbumin in solution. AB - PURPOSE: The purpose of the study was to investigate the relationship of the second virial coefficient, B22, to the extent of irreversible protein aggregation upon storage. METHODS: A monoclonal antibody and ovalbumin were incubated at 37 degrees C (3 months) under various solution conditions to monitor the extent of aggregation. The B22 values of these proteins were determined under similar solution conditions by a modified method of flow-mode static light scattering. The conformation of these proteins was studied using circular dichroism (CD) spectroscopy and second-derivative Fourier transform infrared spectroscopy. RESULTS: Both proteins readily aggregated at pH 4.0 (no aggregation observed at pH 7.4); the extent of aggregation varied with the ionic strength and the presence of cosolutes (sucrose, glycine, and Tween 80). Debye plots of the monoclonal antibody showed moderate attractive interactions at pH 7.4, whereas, at pH 4.0, nonlinear plots were obtained, indicating self-association. CD studies showed partially unfolded structure of antibody at pH 4.0 compared with that at pH 7.4. In the case of ovalbumin, similar B22 values were obtained in all solution conditions irrespective of whether the protein aggregated or not. CD studies of ovalbumin indicated the presence of a fraction of completely unfolded as well as partially unfolded species at pH 4.0 compared with that at pH 7.4. CONCLUSIONS: The formation of a structurally altered state is a must for irreversible aggregation to proceed. Because this aggregation-prone species could be an unfolded species present in a small fraction compared with that of the native state or it could be a partially unfolded state whose net interactions are not significantly different compared with those of the native state, yet the structural changes are sufficient to lead to long-term aggregation, it is unlikely that B22 will correlate with long-term aggregation. PMID- 16715375 TI - Application of micro- and nano-electromechanical devices to drug delivery. AB - Micro- and nano-electromechanical systems (MEMS and NEMS)-based drug delivery devices have become commercially-feasible due to converging technologies and regulatory accommodation. The FDA Office of Combination Products coordinates review of innovative medical therapies that join elements from multiple established categories: drugs, devices, and biologics. Combination products constructed using MEMS or NEMS technology offer revolutionary opportunities to address unmet medical needs related to dosing. These products have the potential to completely control drug release, meeting requirements for on-demand pulsatile or adjustable continuous administration for extended periods. MEMS or NEMS technologies, materials science, data management, and biological science have all significantly developed in recent years, providing a multidisciplinary foundation for developing integrated therapeutic systems. If small-scale biosensor and drug reservoir units are combined and implanted, a wireless integrated system can regulate drug release, receive sensor feedback, and transmit updates. For example, an "artificial pancreas" implementation of an integrated therapeutic system would improve diabetes management. The tools of microfabrication technology, information science, and systems biology are being combined to design increasingly sophisticated drug delivery systems that promise to significantly improve medical care. PMID- 16715376 TI - In vitro studies of intestinal permeability and hepatic and intestinal metabolism of 8-prenylnaringenin, a potent phytoestrogen from hops (Humulus lupulus L.). AB - PURPOSE: The absorption potential and metabolism of 8-prenylnaringenin (8-PN) from hops (Humulus lupulus L.) were investigated. 8-PN is a potent estrogen with the potential to be used for the relief of menopausal symptoms in women. METHODS: Monolayers of the human intestinal epithelial cancer cell line Caco-2 and human hepatocytes were incubated with 8-PN to model its intestinal absorption and hepatic metabolism, respectively. RESULTS: The apparent permeability coefficients for 8-PN in the apical-to-basolateral and basolateral-to-apical directions of a Caco-2 monolayer were 5.2 +/- 0.7 x 10(-5) and 4.9 +/- 0.5 x 10(-5) cm/s, respectively, indicating good intestinal absorption via passive diffusion. Both glucuronide and sulfate conjugates of 8-PN were detected in the Caco-2 cell incubations. The 4'-O-glucuronide was the predominant Caco-2 cell metabolite, followed by 7-O-sulfate and 4'-O-sulfate. Both phase I and phase II metabolites of 8-PN were formed by human hepatocytes. The 7-O-glucuronide was the most abundant hepatocyte metabolite, and no sulfate conjugates were detected. Incubations with various cDNA-expressed UDP-glucuronosyltransferases indicated that the isozymes UGT1A1, UGT1A6, UGT1A8, and UGT1A9 were responsible for glucuronidation of 8-PN. CONCLUSIONS: Although orally administered 8-PN should be readily absorbed from the intestine, its bioavailability should be reduced significantly by intestinal and hepatic metabolism. PMID- 16715377 TI - Brain uptake of nonsteroidal anti-inflammatory drugs: ibuprofen, flurbiprofen, and indomethacin. AB - PURPOSE: To determine the roles of blood-brain barrier (BBB) transport and plasma protein binding in brain uptake of nonsteroidal anti-inflammatory drugs (NSAIDs) ibuprofen, flurbiprofen, and indomethacin. METHODS: Brain uptake was measured using in situ rat brain perfusion technique. RESULTS: [14C]Ibuprofen, [3H]flurbiprofen, and [14C]indomethacin were rapidly taken up into the brain in the absence of plasma protein with BBB permeability-surface area products (PS(u)) to free drug of (2.63 +/- 0.11) x 10(-2), (1.60 +/- 0.08) x 10(-2), and (0.64 +/- 0.05) x 10(-2) mL s(-1) g(-1) (n = 9-11), respectively. BBB [14C]ibuprofen uptake was inhibited by unlabeled ibuprofen (Km = 0.85 +/- 0.02 mM, Vmax = 13.5 +/- 0.4 nmol s(-1) g(-1)) and indomethacin, but not by pyruvate, probenecid, digoxin, or valproate. No evidence was found for saturable BBB uptake of [3H]flurbiprofen or [14C]indomethacin. Initial brain uptake for all three NSAIDs was reduced by the addition of albumin to the perfusion buffer. The magnitude of the brain uptake reduction correlated with the NSAID free fraction in the perfusate. CONCLUSIONS: Free ibuprofen, flurbiprofen, and indomethacin rapidly cross the BBB, with ibuprofen exhibiting a saturable component of transport. Plasma protein binding limits brain NSAID uptake by reducing the free fraction of NSAID in the circulation. PMID- 16715378 TI - Recombinant human serum albumin dimer has high blood circulation activity and low vascular permeability in comparison with native human serum albumin. AB - PURPOSE: Human serum albumin (HSA) is used clinically as an important plasma expander. Albumin infusion is not recommended for critically ill patients with hypovolemia, burns, or hypoalbuminemia because of the increased leakage of albumin into the extravascular spaces, thereby worsening edema. In the present study, we attempted to overcome this problem by producing a recombinant HSA (rHSA) dimer with decreased vascular permeability and an increased half-life. METHODS: Two molecules of rHSA were genetically fused to produce a recombinant albumin dimer molecule. The pharmacokinetics and biodistribution of the recombinant proteins were evaluated in normal rats and carrageenin-induced paw edema mouse model. RESULTS: The conformational properties of this rHSA dimer were similar to those for the native HSA (the HSA monomer), as evidenced by the Western blot and spectroscopic studies. The biological half-life and area under the plasma concentration-time curve of the rHSA dimer were approximately 1.5 times greater than those of the monomer. Dimerization has also caused a significant decrease in the total body clearance and distribution volume at the steady state of the native HSA. rHSA dimer accumulated to a lesser extent in the liver, skin, muscle, and fat, as compared with the native HSA. Up to 96 h, the vascular permeability of the rHSA dimer was less than that of the native HSA in paw edema mouse models. A prolonged plasma half-life of the rHSA dimer was also observed in the edema model rats. CONCLUSIONS: rHSA dimer has a high retention rate in circulating blood and a lower vascular permeability than that of the native HSA. PMID- 16715379 TI - Efficacy of siRNA nanocapsules targeted against the EWS-Fli1 oncogene in Ewing sarcoma. AB - The EWS-Fli1 fusion gene encodes for a chimeric oncogenic transcription factor considered to be the cause of the Ewing sarcoma. The efficiency of small interfering RNAs (siRNAs) targeted toward the EWS-Fli1 transcript (at the junction point type 1) was studied, free or encapsulated into recently developed polyisobutylcyanoacrylate aqueous core nanocapsules. Because this mRNA sequence is only present in cancer cells, it therefore constituted a relevant target. Studies of the intracellular penetration by confocal microscopy in NIH/3T3 EWS Fli1 cells showed that nanocapsules improved the intracellular penetration of siRNA with mainly a cytoplasmic localization. These biodegradable siRNA-loaded nanocapsules were then tested in vivo on a mice xenografted EWS-Fli1-expressing tumor; they were found to trigger a dose-dependant inhibition of tumor growth after intratumoral injection. A specific inhibition of EWS-Fli1 was observed, too. These findings now open new prospects for the treatment of experimental cancers with junction oncogenes. PMID- 16715380 TI - Targeted high lung concentrations of itraconazole using nebulized dispersions in a murine model. AB - PURPOSE: The purpose of this study was to investigate the delivery of itraconazole (ITZ) particles to a murine lung model by nebulization. METHODS: Three ITZ formulations were prepared and characterized in the dry state using contact angle, dissolution, X-ray powder diffraction, scanning electron microscopy, and Brunauer-Emmett-Teller surface area analysis. Aerodynamic particle size distributions and lung deposition studies in 14 outbred male ICR mice were performed using aqueous dispersions of all the formulations. A separate dosing uniformity study was also performed to qualify use of the chamber. RESULTS: All formulations had an aggregated particle size of approximately 30 microm in diameter. Two formulations showed that 80% of the drug dissolved in less than 5 min. The remaining ITZ formulation had a slower dissolution and the lowest total emitted dose from the nebulizer used. High concentrations of ITZ were shown to be present in the mouse lung during the lung deposition study, up to 16.8 +/- 0.13 microg/g (+/- SE) were achieved. Concentrations of up to 0.76 +/ 0.03 microg/g (+/- SE) could be maintained from the single nebulized dose for at least 24 h. CONCLUSION: An effective method of targeted delivery of ITZ to the deep lung is presented that may be useful for the treatment and prevention of acute fungal infections. PMID- 16715381 TI - Synthesis and enhancing effect of transkarbam 12 on the transdermal delivery of theophylline, clotrimazole, flobufen, and griseofulvin. AB - PURPOSE: Dodecyl-6-aminohexanoate (DDEAC) is a transdermal permeation enhancer with excellent activity, low toxicity, and no dermal irritation. We hypothesized that DDEAC reacts with air CO2 to form a two-chain ammonium carbamate- Transkarbam 12 (T12)--which is responsible for the enhancing effect. METHODS: DDEAC and T12 were synthesized, their structures were confirmed by spectral methods, and their enhancing activity was studied using the Franz diffusion cell and human skin. A high-performance liquid chromatography method was developed for determination of T12, and its biodegradability was evaluated using porcine esterase. RESULTS: Only the carbamate salt T12 was responsible for the high enhancing activity; DDEAC tested under argon to avoid reaction with CO2 was inactive. T12 enhanced transdermal permeation of drugs covering a wide range of physicochemical properties, including theophylline (enhancement ratio up to 55.6), clotrimazole (7.7), flobufen (5.0), and griseofulvin (24). The activity was pH-dependent, further confirming the importance of the carbamate structure. The metabolization of T12 followed a second-order kinetics with t(1/2) = 31 min. CONCLUSION: Our results indicate that T12 is a promising biodegradable permeation enhancer for a wide range of drugs, and the structurally novel group of carbamate enhancers warrants further investigation. PMID- 16715382 TI - Comparison of nanogel drug carriers and their formulations with nucleoside 5' triphosphates. AB - PURPOSE: The aim of the study is to synthesize and characterize nanogel carriers composed of amphiphilic polymers and cationic polyethylenimine for encapsulation and delivery of cytotoxic nucleoside analogs 5'-triphosphates (NTPs) into cancer cells. METHODS: Nanogels were synthesized by a novel micellar approach and compared with carriers prepared by the emulsification/evaporation method. Complexes of nanogels with NTP were prepared; particle size and in vitro drug release were characterized. Resistance of the nanogel-encapsulated NTP to enzymatic hydrolysis was analyzed by ion-pair high-performance liquid chromatography. Binding to isolated cellular membranes, cellular accumulation and cytotoxicity were compared using breast carcinoma cell lines CL-66, MCF-7, and MDA-MB-231. In vivo biodistribution of the 3H-labeled NTP encapsulated in different types of nanogels was evaluated in comparison to the injected NTP alone. RESULTS: Nanogels with a particle size of 100-300 nm in the unloaded form and less than 140 nm in the NTP-loaded form were prepared. An in vitro release of NTP was >50% during the first 24 h. Nanogel formulations ensured increased NTP drug stability against enzymatic hydrolysis as compared to the drug alone. Pluronic-based nanogels NG(F68), NG(F127), NG(P85), and NGM(P123) demonstrated 2 2.5 times enhanced interaction with cellular membranes and association with various cancer cells compared to NG(PEG). Among them, NG(F68) and NG(F127) exhibited the lowest cytotoxicity. Injection of nanogel-formulated NTP significantly modulated the drug accumulation in different mouse organs. CONCLUSIONS: Nanogels composed of Pluronic F68 and P123 were shown to display certain advanced properties compared to NG(PEG) as a drug delivery system for NTP analogs. Formulations of nucleoside analogs in active NTP form with these nanogels will improve the delivery of these cytotoxic drugs to cancer cells and the therapeutic potential of this anticancer chemotherapy. PMID- 16715383 TI - Formulation and characterization of lipid-coated tobramycin particles for dry powder inhalation. AB - PURPOSE: This study was conducted to develop and evaluate the physicochemical and aerodynamic characteristics of lipid-coated dry powder formulations presenting particularly high lung deposition. METHODS: Lipid-coated particles were prepared by spray-drying suspensions with different concentrations of tobramycin and lipids. The solid-state properties of the formulations, including particle size and morphology, were assessed by scanning electron microscopy and laser diffraction. Aerosol performance was studied by dispersing the powders into a Multistage Liquid Impinger and determining drug deposition by high-performance liquid chromatography. RESULTS: Particle size distributions of the formulations were unimodal, narrow with more than 90% of the particles having a diameter of less than 2.8 microm. All powder formulations exhibited mass median diameters of less than 1.3 and 3.2 microm, as determined by two different laser diffraction methods, the Malvern's Mastersizer and Spraytec, respectively. The fine particle fraction varied within a range of 50.5 and 68.3%. CONCLUSIONS: Lipid coating of tobramycin formulations resulted in a reduced agglomeration tendency and in high fine particle fraction values, thus improving drug deposition. The very low excipients content (about 5% m/m) of these formulations offers the benefit of delivering particularly huge concentrations of antibiotic directly to the site of infection, while minimizing systemic exposure, and may provide a valuable alternative treatment of cystic fibrosis. PMID- 16715384 TI - Chitosan-modified dry powder formulations for pulmonary gene delivery. AB - PURPOSE: Spray-drying is an effective process for preparing micron-dimensioned particles for pulmonary delivery. Previously, we have demonstrated enhanced dispersibility and fine particle fraction of spray-dried nonviral gene delivery formulations using amino acids or absorption enhancers as dispersibility enhancing excipients. In this study, we investigate the use of the cationic polymer chitosan as a readily available and biocompatible dispersibility enhancer. METHODS: Lactose-lipid:polycation:pDNA (LPD) powders were prepared by spray-drying and post-mixed with chitosan or spray-dried chitosan. In addition, the water-soluble chitosan derivative, trimethyl chitosan, was added to the lactose-LPD formulation before spray-drying. RESULTS: Spray-dried chitosan particles, displaying an irregular surface morphology and diameter of less than 2 microm, readily adsorbed to lactose-LPD particles following mixing. In contrast with the smooth spherical surface of lactose-LPD particles, spray-dried trimethyl chitosan-lactose-LPD particles demonstrated increased surface roughness and a unimodal particle size distribution (mean diameter 3.4 microm), compared with the multimodal distribution for unmodified lactose-LPD powders (mean diameter 23.7 microm). The emitted dose and in vitro deposition of chitosan-modified powders was significantly greater than that of unmodified powders. Moreover, the inclusion of chitosan mediated an enhanced level of reporter gene expression. CONCLUSIONS: In summary, chitosan enhances the dispersibility and in vitro pulmonary deposition performance of spray-dried powders. PMID- 16715385 TI - A novel in vitro percutaneous penetration model: evaluation of barrier properties with p-aminobenzoic acid and two of its derivatives. AB - PURPOSE: The objective of this study was to evaluate the utility of a stratum corneum substitute (SCS) as a novel in vitro percutaneous penetration model. The SCS consists of synthetic stratum corneum (SC) lipids (cholesterol, free fatty acids, and specific ceramides) applied onto a porous substrate. The composition, organization, and orientation of lipids in the SCS bear high resemblance to that of the intercellular barrier lipids in SC. METHODS: The barrier integrity of the SCS was evaluated by means of passive diffusion studies, using three model compounds with different lipophilicities. The effects of lipid layer thickness, permeant lipophilicity, and altered lipid composition on the barrier properties were investigated, using isolated human SC as a control sample. RESULTS: For all three model compounds, the permeability characteristics of the SCS with a 12-mum thick lipid layer closely resemble those of human SC. Modification of the lipid composition, generating an SCS that lacks the characteristic long periodicity phase as present in SC, was accompanied by a 2-fold increased permeability. CONCLUSIONS: The SCS offers an attractive tool to predict solute permeation through human skin. Moreover, as its lipid composition can be modified, they may also serve as a suitable screening model for diseased skin. PMID- 16715386 TI - Beta-relaxation of insulin molecule in lyophilized formulations containing trehalose or dextran as a determinant of chemical reactivity. AB - PURPOSE: The purpose of this study was to elucidate whether the degradation rate of insulin in lyophilized formulations is determined by matrix mobility, as reflected in glass transition temperature (Tg), or by beta-relaxation, as reflected in rotating-frame spin-lattice relaxation time (T1rho). METHODS: The storage stability of insulin lyophilized with dextran was investigated at various relative humidities (RH; 12-60%) and temperatures (40-90 degrees C) and was compared with previously reported data for insulin lyophilized with trehalose. Insulin degradation was monitored by reverse-phase high-performance liquid chromatography. Furthermore, the T1rho of the insulin carbonyl carbon in the lyophilized insulin-dextran and insulin-trehalose systems was measured at 25 degrees C by 13C solid-state NMR, and the effect of trehalose and dextran on T1rho was compared at various humidities. RESULTS: The degradation rate of insulin lyophilized with dextran was not significantly affected by the Tg of the matrix, even at low humidity (12% RH), in contrast to that of insulin lyophilized with trehalose. The insulin-dextran system exhibited a substantially greater degradation rate than the insulin-trehalose system at a given temperature below the Tg. The difference in degradation rate between the insulin-dextran and insulin-trehalose systems observed at 12% RH was eliminated at 43% RH. In addition, the T1rho of the insulin carbonyl carbon at low humidity (12% RH) was prolonged by the addition of trehalose, but not by the addition of dextran. This difference was eliminated at 23% RH, at which point the solid remained in the glassy state. These findings suggest that the beta-relaxation of insulin is inhibited by trehalose at low humidity, presumably as a result of insulin trehalose interaction, and thus becomes a rate determinant. In contrast, dextran, whose ability to interact with insulin is thought to be less than that of trehalose, did not inhibit the beta-relaxation of insulin, and thus, the chemical activational barrier (activation energy) rather than beta-relaxation becomes the major rate determinant. CONCLUSIONS: Beta-relaxation rather than matrix mobility seems to be more important in determining the stability of insulin in the glassy state in lyophilized formulations containing trehalose and dextran. PMID- 16715387 TI - Structural relaxation of acetaminophen glass. AB - PURPOSE: The aim is to determine the structural stability of acetaminophen glass with time and temperature change, and to examine the merits of adapting the structural relaxation models of the glassy state for pharmaceuticals. METHODS: Differential scanning calorimetry technique has been used to study the acetaminophen glass after keeping the samples for various periods at fixed temperatures and after keeping at various temperatures for fixed periods. RESULTS: A general formalism for thermodynamic changes during storage in a temperature fluctuating environment is given and the kinetics of the enthalpy and entropy decrease determined. At a fixed temperature, the decrease occurs according to a non-exponential kinetics. For the same storage time, but at different temperatures, the enthalpy and entropy decrease rises to a maximum value at a certain temperature and then declines. The peak appears at the temperature at which the internally equilibrated state of the sample is reached for a fixed storage time. The change in the normalized heat capacity during the heating of acetaminophen has been analysed in terms of a non-exponential, non linear enthalpy relaxation model. CONCLUSION: A single set of parameters that fit the data for unannealed acetaminophen glass does not fit the calorimetric data for annealed glass. Since acetaminophen molecules form intermolecular hydrogen bonds in the crystal state and likely to form such bonds more easily in the disordered state, effect of such bonds on structural relaxation is likely to be significant. PMID- 16715388 TI - Characterization of complexes between naftifine and cyclodextrins in solution and in the solid state. AB - Naftifine (NF) is an antifungal drug poorly soluble in basic aqueous solutions. Complexation with cyclodextrins (CDs) improves the physicochemical characteristics of many drugs. The aim of this work is to characterize the interactions between NF and alpha-CD, beta-CD, hydroxypropylbeta-CD, methylbeta CD, and gamma-CD. The studies have been developed in pH 12 aqueous solutions at 25 degrees C and in the solid state. The apparent stability constants of the complexes have been determined from phase-solubility diagrams. In the solid state, crystalline and amorphous complexes have been characterized using X-ray diffraction patterns, thermal analysis, and Fourier transform infrared spectroscopy. The solubility of NF improves with all the CDs studied, with the exception of alpha-CD. Different types of diagrams have been found depending on the CD used. The interaction between NF and hydroxypropylbeta-CD is stronger than that with beta-CD due to the specific properties of the substituents. The coevaporation method can be said the best method in preparing the solid complexes, except for NF-alpha-CD; again, there is no evidence of complexation. Furthermore, the presence of different types of CD structures upon complexation (i.e., cage or channel) has been discussed. Dissolution rate studies have been performed, and a positive influence of complexation in the solid state has been observed. PMID- 16715389 TI - Formulation of solid dosage forms to overcome gastric pH interaction of the factor Xa inhibitor, BMS-561389. AB - PURPOSE: The purpose of the study was to investigate the specific mechanism by which elevated gastric pH reduces the absorption of BMS-561389, a factor Xa inhibitor, and to develop a solid formulation strategy to overcome this gastric pH interaction. METHODS: A dissolution method in an acetate buffer at pH 5.5 was used to evaluate the dissolution behavior of the tablet formulation. A precipitation model was used to screen different excipients for their potential to minimize the pH-dependent absorption of BMS-561389. Excipients that showed promise in the precipitation model were incorporated in modified tablet formulations. Dissolution rate of the modified tablets was also determined by the acetate buffer method. A canine model for pH-dependent absorption was subsequently used to evaluate the tablet formulations. RESULTS: Dissolution studies suggested that the reduced absorption of the original formulation was the result of the precipitation of the poorly water-soluble free base during the initial dissolution of the salt. Modified tablets containing organic acids, sulfobutylether-beta-cyclodextrin, or povidone showed enhanced dissolution as compared with the original formulation. Drug absorption from the tablet containing tartaric acid was substantially independent of gastric pH in the canine model. CONCLUSION: A multitier approach was successful in identifying a solid dosage form that minimizes the pH-dependent absorption of this drug candidate. PMID- 16715390 TI - Insights into the role of electrostatic forces on the behavior of dry pharmaceutical particulate systems. AB - PURPOSE: Under different charging conditions, particles can be either attracted or repulsed by each other, causing powder agglomeration or segregation. Such behavior can be detrimental in many processes aimed at achieving particulate mixture homogeneity. Consequently, the effects of electrostatic charges on mixing kinetics must be well understood to insure a high level of process control, product quality, and reproducibility. METHODS: In Part 1, an electrostatic charger is used to evaluate the ability of the studied particles to develop and retain an induced charge at the surface for a fixed period of time. Part 2 assesses the natural electrostatic charge developed by powders sliding across a stainless steel, plastic, or Tyvek chute. In Part 3, 2 binary systems were formed according to an experimental design under different charging conditions, and their behavior was quantified by measuring the degree of agglomeration attained. RESULTS: This work has shown that each raw material develops a different charge according to its physico-chemical properties and the type of contact surface. Electrostatic charges influence the creation of agglomerates under certain conditions. CONCLUSIONS: The presence of electrostatic charges must be accounted for in any effort to maximize mixing efficiency. PMID- 16715391 TI - Polymer microneedles for controlled-release drug delivery. AB - PURPOSE: As an alternative to hypodermic injection or implantation of controlled release systems, this study designed and evaluated biodegradable polymer microneedles that encapsulate drug for controlled release in skin and are suitable for self-administration by patients. METHODS: Arrays of microneedles were fabricated out of poly-lactide-co-glycolide using a mold-based technique to encapsulate model drugs--calcein and bovine serum albumin (BSA)--either as a single encapsulation within the needle matrix or as a double encapsulation, by first encapsulating the drug within carboxymethylcellulose or poly-L: -lactide microparticles and then encapsulating drug-loaded microparticles within needles. RESULTS: By measuring failure force over a range of conditions, poly-lactide-co glycolide microneedles were shown to exhibit sufficient mechanical strength to insert into human skin. Microneedles were also shown to encapsulate drug at mass fractions up to 10% and to release encapsulated compounds within human cadaver skin. In vitro release of calcein and BSA from three different encapsulation formulations was measured over time and was shown to be controlled by the encapsulation method to achieve release kinetics ranging from hours to months. Release was modeled using the Higuchi equation with good agreement (r2 > or = 0.90). After microneedle fabrication at elevated temperature, up to 90% of encapsulated BSA remained in its native state, as determined by measuring effects on primary, secondary, and tertiary protein structure. CONCLUSIONS: Biodegradable polymer microneedles can encapsulate drug to provide controlled-release delivery in skin for hours to months. PMID- 16715393 TI - The in vitro plasma distribution of a novel cholesteryl ester transfer protein inhibitor, torcetrapib, is influenced by differences in plasma lipid concentrations. AB - PURPOSE: To determine the lipoprotein distribution of Torcetrapib in normolipidemic or hyperlipidemic human plasma and assess any changes in distribution due to lipid profile. METHODS: Torcetrapib was incubated with human plasma samples, and the distribution was measured across four fractions: triglyceride-rich lipoprotein (TRL), low-density lipoprotein, high-density lipoprotein, and lipoprotein-deficient plasma fraction. Two stocks of human plasma were used, one considered normolipidemic (total cholesterol concentration = 164 mg/dL, triglycerides concentration = 139 mg/dL, protein concentration = 912 mg/dL), the other hyperlipidemic (total cholesterol = 260 mg/dL, triglycerides = 775 mg/dL, protein = 917 mg/dL). The plasma samples were incubated with Torcetrapib at 37 degrees C, and the incubation was stopped with the addition of sodium bromide and cooling to 4 degrees C. The plasma samples were then separated by density gradient ultracentrifugation to their lipoprotein fractions. The resulting lipoprotein fractions and an aliquot of incubated plasma were analyzed by a validated gas chromatography/tandem mass spectrometry analytical method. The distribution of Torcetrapib was determined first with varying incubation times, then with several concentrations. RESULTS: At concentrations of 250 and 500 ng/mL, Torcetrapib distributed evenly across the four fractions in normolipidemic plasma. At the same concentrations in hyperlipidemic plasma, approximately 84% of Torcetrapib was found in the TRL fraction, with the remaining 16% evenly partitioned between the low-density lipoprotein, high-density lipoprotein, and lipoprotein-deficient plasma fractions. CONCLUSIONS: The results suggest that lipid profile affects the distribution of Torcetrapib in hyperlipidemic human plasma lipoprotein fractions. The preferential distribution of Torcetrapib into the TRL fraction in hyperlipidemic plasma needs to be investigated to see if it will affect the pharmacological effect of Torcetrapib in vivo. PMID- 16715392 TI - Role of phospholipid transfer protein on the plasma distribution of amphotericin B following the incubation of different amphotericin B formulations. AB - PURPOSE: The purpose of this study was to investigate the role of phospholipid transfer protein (PLTP) on the plasma distribution of amphotericin B (AmpB) following incubation with different AmpB formulations in human plasmas with varying lipid profiles. METHODS: In a first set of experiments, plasma distribution profiles of AmpB were determined following the incubation of Fungizone and lipid-based formulations (Abelcet and AmBisome) at a concentration of 20 microg AmpB/mL for 5-120 min at 37 degrees C in the plasma obtained from six different individuals (total cholesterol concentrations range between 62 and 332 mg/dL). In a second set of experiments, Abelcet, and AmBisome at a concentration of 20 microg AmpB/mL were incubated for 5 min at 37 degrees C in human plasma (total cholesterol = 163 mg/dL) that had been pretreated with an antibody raised up against PLTP (1:400 v/v dilution from stock solution) for 20 min at 37 degrees C. Following incubation, the human plasma was separated into its lipoprotein and lipoprotein-deficient fractions by density gradient ultracentrifugation and analyzed for AmpB content by high-performance liquid chromatography. RESULTS: The majority of AmpB was covered in the lipoprotein deficient plasma and high-density lipoprotein (HDL) fractions following incubation of Fungizone in human plasma. The majority of AmpB (48.7-87.2%) was recovered in the HDL fraction following incubation of Abelcet and AmBisome in human plasma. The presence of the PLTP antibody resulted in a 20% decrease in the percentage AmpB recovered in the HDL fraction following the incubation of Abelcet. However, the plasma distribution of AmpB remained unchanged following the incubation of AmBisome in plasma containing the PLTP antibody. CONCLUSIONS: Taken together, these findings suggest indirect evidence that PLTP may play an important role in the plasma distribution profile of AmpB following the incubation of Abelcet and may be one of the factors responsible for the preferential association of AmpB with HDL when administered as Abelcet. PMID- 16715394 TI - Biopharmaceutical characterization of the telomerase inhibitor BRACO19. AB - PURPOSE: To characterize the telomerase inhibitor and G-quadruplex stabilizing substance 9-[4-(N,N-dimethylamino)phenylamino]-3,6-bis (3-pyrrolodino propionamido) acridine x 3HCl (BRACO19) in terms of biopharmaceutical properties such as solubility, protein binding, interaction with membrane lipids, cytotoxicity, and permeability across pulmonary epithelial cells. METHODS: Protein binding and interaction with membrane lipids were investigated by two high-performance liquid chromatography methods with immobilized human serum albumin and immobilized phosphatidylcholine, respectively. Cytotoxicity (methyl thiazolyl-tetrazolium assay) and transport studies were performed with the bronchial cell lines 16HBE14o- and Calu-3, primary human alveolar epithelial cells, and the intestinal cell line Caco-2. Transport experiments were also done in the presence of cyclosporin A (10 microM) and tetraethylammonium chloride (5 mM) and at low temperature (4 degrees C). RESULTS: BRACO19 has good solubility of at least 2 mg/mL in water and in physiological buffers of pH 7.4 and below. Protein binding to human serum albumin was 38%. No interaction with membrane lipids could be found. Cytotoxicity in 16HBE14o-, Calu-3, and human alveolar epithelial cells was in the range of IC50 = 3.5 to 13.5 microM. Caco-2 cells were not affected at concentrations up to 50 microM. No transport of BRACO19 was detected across either cell monolayer in absorptive direction. In secretory direction, permeability was very low, with P (app) values in the range of 0.25 x 10(-7) to 0.98 x 10(-7) cm/s for all epithelial cell cultures tested. The transport was not influenced by cyclosporin A or tetraethylammonium chloride or at 4 degrees C, indicating that no efflux/influx systems or active transport are involved. CONCLUSIONS: From these results, we conclude that the very poor permeability of BRACO19 is its main biopharmaceutical limitation. Further applications will require a suitable formulation to warrant adequate delivery across cellular barriers. PMID- 16715396 TI - Solid-state characterization of buspirone hydrochloride polymorphs. AB - The purpose of this study was to characterize Form 1 and Form 2 of buspirone hydrochloride, an anxiolytic medicine. The techniques used for characterization included microscopy (optical, hot stage, and scanning electron microscopy), thermal analysis (differential scanning calorimetry and thermogravimetric analysis), solid-state Fourier transform infrared (FTIR) spectroscopy, X-ray powder diffractometry (XRPD), and Raman spectroscopy. Morphologically, Form 1 and Form 2 consist of plate and columnar crystals, respectively, with good filterability. Thermal analysis showed that the two forms are enantiotropic over the studied temperature range. The FTIR method was used successfully for the quantification of Form 1 in a mixture of Forms 1 and 2. The ratio of a characteristic peak to a reference peak and the chemometric method were used to obtain the calibration curve. The Raman peak shifts showed the difference between the two forms especially for the n-butyl group. The large number of distinguishable XRPD peaks in the region of 5 degrees to 30 degrees 2theta of the two polymorphs demonstrated that XRPD is a useful tool for quantitative and qualitative analysis of polymorphs. PMID- 16715395 TI - Biantennary glycans as well as genetic variants of alpha1-acid glycoprotein control the enantioselectivity and binding affinity of oxybutynin. AB - PURPOSE: The purpose of this study was to investigate the role of biantennary branching glycans of alpha1-acid glycoprotein (AGP) and its genetic variants in the enantioselective binding of oxybutynin (OXY). METHOD: Human native AGP was separated using imminodiacetate-copper (II) affinity chromatography into two fractions, the A variant and a mixture of the F1 and S variants (F1-S). These fractionated AGPs were further separated by concanavalin A affinity chromatography into two fractions, with and without biantenarry glycans. An on line high-performance liquid chromatography (HPLC) system consisting of a high performance frontal analysis column, an extraction column, and an analytical HPLC column was developed to determine the binding affinities of OXY enantiomers for respective AGP species. RESULTS: The total binding affinity as well as the enantiomeric selectivity of OXY in the F1-S mixed variant was significantly higher than that for the A variant, indicating that the chiral recognition ability of native AGP for the OXY enantiomers highly depends on the F1-S mixed variant. Furthermore, not only the genetic variants but also bianntenary glycans of AGP affect the binding affinity of OXY and are also responsible for the enantioselectivity. CONCLUSIONS: Both genetic variants and glycan structures significantly contribute to the enantioselectivity and the binding affinity of OXY. PMID- 16715399 TI - Anaerobic bioremediation of groundwater containing a mixture of 1,1,2,2 tetrachloroethane and chloroethenes. AB - This study investigated the biotransformation pathways of 1,1,2,2 tetrachloroethane (1,1,2,2-TeCA) in the presence of chloroethenes (i.e. tetrachloroethene, PCE; trichloroethene, TCE) in anaerobic microcosms constructed with subsurface soil and groundwater from a contaminated site. When amended with yeast extract, lactate, butyrate, or H2 and acetate, 1,1,2,2-TeCA was initially dechlorinated via both hydrogenolysis to 1,1,2-trichloroethane (1,1,2-TCA) (major pathway) and dichloroelimination to dichloroethenes (DCEs) (minor pathway), with both reactions occurring under sulfidogenic conditions. In the presence of only H2, the hydrogenolysis of 1,1,2,2-TeCA to 1,1,2-TCA apparently required the presence of acetate to occur. Once formed, 1,1,2-TCA was degraded predominantly via dichloroelimination to vinyl chloride (VC). Ultimately, chloroethanes were converted to chloroethenes (mainly VC and DCEs) which persisted in the microcosms for very long periods along with PCE and TCE originally present in the groundwater. Hydrogenolysis of chloroethenes occurred only after highly reducing methanogenic conditions were established. However, substantial conversion to ethene (ETH) was observed only in microcosms amended with yeast extract (200 mg/l), suggesting that groundwater lacked some nutritional factors which were likely provided to dechlorinating microorganisms by this complex organic substrate. Bioaugmentation with an H2-utilizing PCE-dechlorinating Dehalococcoides spp. -containing culture resulted in the conversion of 1,1,2,2 TeCA, PCE and TCE to ETH and VC. No chloroethanes accumulated during degradation suggesting that 1,1,2,2-TeCA was degraded through initial dichloroelimination into DCEs and then typical hydrogenolysis into ETH and VC. PMID- 16715398 TI - Mechanisms underlying overdrive suppression and overdrive excitation in guinea pig sino-atrial node. AB - The hypothesis that the pause that follows overdrive of the sino-atrial node (SAN) might be the net result of overdrive excitation and overdrive suppression was tested by studying rate and force patterns induced by overdrive in isolated guinea pig SAN superfused in vitro. In Tyrode solution, the pause is short and changes but little with longer or faster drives. In high [K(+)](o) solution, longer overdrives increase force percent-wise more than in Tyrode solution, shorten the pause and are followed by greater rate and force. When the SAN (quiescent in high [K(+)](o)) is driven at 6/min, faster overdrives are followed by stronger, slowly decreasing contractions. Alternating 10 s drives with 10 s pauses have little effect on force and rate in Tyrode solution, but progressively increase force and rate in high [K(+)](o). Cesium has effects similar to high [K(+)](o). High [Ca(2+)](o) increases force and in high [K(+)](o) increases the rate as well as it shortens the pause, whereas Ni(2+) decreases force as well as rate and lengthens the pause. Barium dissociates the effects on force and rate. Lidocaine and tetrodotoxin decrease rate and force, and increase the pause duration. In overdrive excitation, the increase in rate is associated with an enhancement of diastolic voltage oscillations. It is concluded that in SAN the prevalence of Ca(2+) load leads to overdrive excitation whereas the prevalence of Na(+) load leads to overdrive suppression. In Tyrode solution, the pause after drive appears to be the net result of these two different mechanisms. PMID- 16715400 TI - The impact of bioaugmentation on metal cyanide degradation and soil bacteria community structure. AB - Metal cyanides are significant contaminants of many soils found at the site of former industrial activity. In this study we isolated bacteria capable of degrading ferric ferrocyanide and K2Ni(CN)4. One of these bacteria a Rhodococcus spp. was subsequently used to bioaugment a minimal medium broth, spiked with K2Ni(CN)4, containing 1 g of either an uncontaminated topsoil or a former coke works site soil. Degradation of the K2Ni(CN)4 was observed in both soils, however, bioaugmentation did not significantly impact the rate or degree of K2Ni(CN)4 removal. Statistical analysis of denaturing gradient gel electrophoresis profiles showed that the topsoil bacterial community had a higher biodiversity, and its structure was not significantly affected by either K2Ni(CN)4 or bioaugmentation. In contrast, profiles from the coke works site indicated significant changes in the bacterial community in response to these additions. Moreover, in both soils although bioaugmentation did not affect rates of biodegradation the Rhodococcus spp. did become established in the communities in broths containing both top and coke works soil. We conclude that bacterial communities from contaminated soils with low biodiversity are much more readily perturbed through interventions such as contamination events or bioaugmentation treatments and discuss the implications of these findings for bioremediation studies. PMID- 16715402 TI - Impacts of bioremediation schemes on bacterial population in naphthalene contaminated marine sediments. AB - Microcosm experiments were conduced in which the surface of marine sediment was contaminated with naphthalene and subjected to either of three different bioremediation schemes, i.e., biostimulation (BS) by supplementing with slow release nitrogen and phosphorus fertilizers, bioaugmentation (BA) by inoculating with Cycloclasticus sp. E2, an aromatics-degrading bacterium identified to play an important role for aromatic-hydrocarbon degradation in marine environments and combination (CB) of BS and BA. These three schemes were found to be similarly effective for removing naphthalene, while naphthalene disappearance in sediment without any treatment (WT) was slower than those in the treated sediments. Shifts in bacterial populations during and after bioremediation were analyzed by denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA gene fragments. It was found that the Cycloclasticus rRNA type occurred as the strongest bands in the course of naphthalene degradation. Clustering analysis of DGGE profiles showed that bacterial populations in the WT, BS and CB sediments differed consistently from those in the uncontaminated control, while the profile for the BA sediment was finally included in the cluster for uncontaminated control sediments after a 150-day treatment. The results suggest that bioaugmentation with ecologically competent pollutant-degrading bacteria is an ecologically promising bioremediation scheme. PMID- 16715401 TI - Isolation and characterization of a pseudomonas oleovorans degrading the chloroacetamide herbicide acetochlor. AB - To date, no pure bacterial cultures that could degrade acetochlor have been described. In this study, one strain of microorganism capable of degrading acetochlor, designated as LCa2, was isolated from acetochlor-contaminated soil. The strain LCa2 is Pseudomonas oleovorans according to the criteria of Bergey's manual of determinative bacteriology and sequence analysis of the partial 16S rRNA gene. Optimum growth temperature and pH were 35 degrees C and 8.0, respectively. The strain could degrade 98.03% of acetochlor treated at a concentration of 7.6 mg l(-1) after 7 days of incubation and could tolerate 200 mg l(-1) of acetochlor. When the acetochlor concentration became higher, the degradation cycle became longer. The acetochlor biodegradation products were identified by GC-MS based on mass spectral data and fragmentation patterns. The main plausible degradative pathways involved dechlorination, hydroxylation, N dealkylation, C-dealkylation and dehydrogenation. PMID- 16715403 TI - The ACCEL model for accelerating the detoxification kinetics of hydrocarbons requiring initial monooxygenation reactions. AB - The two-tank accelerator/aerator modification of activated sludge significantly increases the biodegradation of hydrocarbons requiring initial monooxygenation reactions, such as phenol and 2,4-dichlorophenol (DCP). The small accelerator tank has a controlled low dissolved oxygen (DO) concentration that can enrich the biomass in NADH + H+. It also has a very high specific growth rate (mu acc) that up-regulates the biomass's content of the monooxygenase enzyme. Here, we develop and test the ACCEL model, which quantifies all key phenomena taking place when the accelerator/aerator system is used to enhance biodegradation of hydrocarbons requiring initial monooxygenations. Monooxygenation kinetics follow a multiplicative relationship in which the organic substrates (phenol or DCP) and DO have separate Monod terms, while the biomass's content of NADH + H+ has a first-order term. The monooxygenase enzyme has different affinities (K values) for phenol and DCP. The biomass's NADH + H+ content is based on a proportioning of NAD(H) according to the relative rates of NADH + H+ sources and sinks. Biomass synthesis occurs simultaneously through utilization of acetate, phenol, and DCP, but each has its own true yield. The ACCEL model accurately simulates all trends for one-tank and two-tank experiments in which acetate, phenol, and DCP are biodegraded together. In particular, DCP removal is affected most by DOacc and the retention-time ratio, Theta acc/Theta total. Adding an accelerator tank dramatically increases DCP removal, and the best DCP removal occurs for 0.2 < DOacc < 0.5 mg/l and 0.08 < Theta acc/Theta total < 0.2. The rates of phenol and DCP utilization follow the multiplicative relationship with a maximum specific rate coefficient proportional to mu acc. Finally, mu acc increases rapidly for Theta acc/Theta total < 0.25, acetate removal in the accelerator fuels the high mu acc, and the biomass's NADH + H+ content increases very dramatically for DO acc < 0.25 mg/l. PMID- 16715404 TI - Bioremediation by composting of heavy oil refinery sludge in semiarid conditions. AB - The present work attempts to ascertain the efficacy of low cost technology (in our case, composting) as a bioremediation technique for reducing the hydrocarbon content of oil refinery sludge with a large total hydrocarbon content (250-300 g kg(-1)), in semiarid conditions. The oil sludge was produced in a refinery sited in SE Spain The composting system designed, which involved open air piles turned periodically over a period of 3 months, proved to be inexpensive and reliable. The influence on hydrocarbon biodegradation of adding a bulking agent (wood shavings) and inoculation of the composting piles with pig slurry (a liquid organic fertiliser which adds nutrients and microbial biomass to the pile) was also studied. The most difficult part during the composting process was maintaining a suitable level of humidity in the piles. The most effective treatment was the one in which the bulking agent was added, where the initial hydrocarbon content was reduced by 60% in 3 months, compared with the 32% reduction achieved without the bulking agent. The introduction of the organic fertiliser did not significantly improve the degree of hydrocarbon degradation (56% hydrocarbon degraded). The composting process undoubtedly led to the biodegradation of toxic compounds, as was demonstrated by ecotoxicity tests using luminescent bacteria and tests on plants in Petri dishes. PMID- 16715405 TI - 2,4-Dichlorophenoxyacetic acid (2,4-D) utilization by Delftia acidovorans MC1 at alkaline pH and in the presence of dichlorprop is improved by introduction of the tfdK gene. AB - Growth of Delftia acidovorans MC1 on 2,4-dichlorophenoxyacetic acid (2,4-D) and on racemic 2-(2,4-dichlorophenoxy)propanoic acid ((RS)-2,4-DP) was studied in the perspective of an extension of the strain's degradation capacity at alkaline pH. At pH 6.8 the strain grew on 2,4-D at a maximum rate (mu max) of 0.158 h(-1). The half-maximum rate-associated substrate concentration (Ks) was 45 microM. At pH 8.5 mu max was only 0.05 h(-1) and the substrate affinity was mucher lower than at pH 6.8. The initial attack of 2,4-D was not the limiting step at pH 8.5 as was seen from high dioxygenase activity in cells grown at this pH. High stationary 2,4-D concentrations and the fact that mu max with dichlorprop was around 0.2 h( 1) at both pHs rather pointed at limited 2,4-D uptake at pH 8.5. Introduction of tfdK from D. acidovorans P4a by conjugation, coding for a 2,4-D-specific transporter resulted in improved growth on 2,4-D at pH 8.5 with mu max of 0.147 h(-1) and Ks of 267 microM. Experiments with labeled substrates showed significantly enhanced 2,4-D uptake by the transconjugant TK62. This is taken as an indication of expression of the tfdK gene and proper function of the transporter. The uncoupler carbonylcyanide m-chlorophenylhydrazone (CCCP) reduced the influx of 2,4-D. At a concentration of 195 microM 2,4-D, the effect amounted to 90% and 50%, respectively, with TK62 and MC1. Cloning of tfdK also improved the utilization of 2,4-D in the presence of (RS)-2,4-DP. Simultaneous and almost complete degradation of both compounds occurred in TK62 up to D = 0.23 h(-1) at pH 6.8 and up to D = 0.2 h(-1) at pH 8.5. In contrast, MC1 left 2,4-D largely unutilized even at low dilution rates when growing on herbicide mixtures at pH 8.5. PMID- 16715406 TI - Isolation of fenitrothion-degrading strain Burkholderia sp. FDS-1 and cloning of mpd gene. AB - A short rod shaped, gram-negative bacterium strain Burkholderia sp. FDS-1 was isolated from the sludge of the wastewater treating system of an organophosphorus pesticides manufacturer. The isolate was capable of using fenitrothion as the sole carbon source for its growth. FDS-1 first hydrolyzed fenitrothion to 3 methyl-4-nitrophenol, which was further metabolized to nitrite and methylhydroquinone. The addition of other carbon source and omitting phosphorus source had little effect on the hydrolysis of fenitrothion. The gene encoding the organophosphorus hydrolytic enzyme was cloned and sequenced. The sequence was similar to mpd, a gene previously shown to encode a parathion-methyl-hydrolyzing enzyme in Plesiomonas sp. M6. The inoculation of strain FDS-1 (10(6) cells g(-1)) to soil treated with 100 mg fenitrothion emulsion kg(-1) resulted in a higher degradation rate than in noninoculated soils regardless of the soil sterilized or nonsterilized. These results highlight the potential of this bacterium to be used in the cleanup of contaminated pesticide waste in the environment. PMID- 16715407 TI - Efforts to explain and control the prolonged thermophilic period in two-phase olive oil mill sludge composting. AB - The aim of this paper was to evaluate the use of different bulking agents in different ratios as a means to control, optimise and eventually reduce the duration of the thermophilic period in two-phase olive oil mill sludge (OOMS) composting. The bulking agents used were: (i) olive tree leaves (OTL), (ii) olive tree shredded branches (OTB) and (iii) woodchips (WDC). The selection of these materials was based on their abundance and availability on the island of Crete, the southernmost point of Greece. The ratios studied were: Pile 1, OOMS:OTL in 1:1 v/v; Pile 2, OOMS:WDC in 1:1.5 v/v; Pile 3, OOMS:OTL in 1:2 v/v; Pile 4, OOMS:OTL:OTB in 1:1:1 v/v; and Pile 5, OOMS:OTL:OTB in 1:1:2 v/v. The composting system used was that of windrows with the volume of each pile approximately 20-25 m3. The experiments took place over two consecutive years. A composting turner was used and turnings were performed at one and two week intervals. In each pile a variety of physiochemical parameters were monitored. Temperature remained high in all five trials. Piles 1, 2, 3, 4 and 5 temperatures recorded values of above 50 degrees C for 106, 158, 160, 175 and 183 days, respectively. Volumes were reduced by approximately 67%, 62%, 63%, 80% and 84%, respectively. Temperature remained high, mainly due to the presence in large amounts of oily substances which during their complete oxidation release important amounts of energy and aid the cometabolism of more stable molecules such as lignin. This process is better described as the slow "burning" of a "fuel" mixture in an "engine" than composting. This approach is based on the extensive similarities of this process to that of crude oil sludge or similar waste composting. PMID- 16715408 TI - Physiological roles of glycerol-transporting aquaporins: the aquaglyceroporins. AB - A subclass of aquaporin (AQP) water channels, termed aquaglyceroporins, are also able to transport glycerol and perhaps urea and other small solutes. Although extensive data exist on the physiological roles of aquaporin-facilitated water transport, until recently the biological significance of glycerol transport by the mammalian aquaglyceroporins has been unknown. There is now compelling evidence for involvement of aquaglyceroporin- facilitated glycerol transport in skin hydration and fat cell metabolism. Mice deficient in AQP3 have dry skin, reduced skin elasticity and impaired epidermal biosynthesis. Mice lacking AQP7 manifest progressive adipocyte fat accumulation and hypertrophy. These skin and fat phenotypes are attributable to impaired glycerol transport. A potential implication of these findings is the possibility of modulation of aquaglyceroporin expression or function in the therapy of skin diseases and obesity. PMID- 16715409 TI - Overcoming HIV drug resistance through rational drug design based on molecular, biochemical, and structural profiles of HIV resistance. AB - There are 20 available drugs for the treatment of human immunodeficiency virus (HIV) infection. With a single exception, all of these drugs inhibit either HIV reverse transcriptase or protease. Reverse transcriptase inhibitors can be further categorized as nucleoside/nucleotide analogs or non-nucleoside reverse transcriptase inhibitors. Resistance that has emerged against all available antiretroviral drugs represents a major challenge in the therapy of HIV infection. Nevertheless, extensive analysis of the molecular and structural mechanisms by which such mutations confer resistance has accumulated over the years. This understanding has driven the development and refinement of novel compounds capable of maintaining antiviral activity against both wild-type and drug-resistant HIV strains. The molecular, biochemical, and structural profiles of reverse transcriptase inhibitor and protease inhibitor resistance are discussed. In addition, how this knowledge has been utilized to generate a new generation of antiviral drugs with activity against drug-resistant HIV is reviewed. PMID- 16715411 TI - Plant thionins--the structural perspective. AB - Thionins belong to a rapidly growing family of biologically active peptides in the plant kingdom. Thionins are small ( approximately 5 kDA), cysteine-rich peptides with toxic and antimicrobial properties. They show a broad cellular toxicity against wide range of organisms and eukaryotic cell lines; while possessing some selectivity. Thionins are believed to be involved in protection against plant pathogens, including bacteria and fungi, by working directly at the membrane. The direct mechanism of action is still surrounded by controversy. Here the results of structural studies are reviewed and confronted with recent results of biophysical studies aimed at defining the function of thionins. The proposed toxicity mechanisms are reviewed and the attempt to reconcile competing hypotheses with a wealth of structural and functional studies is made. PMID- 16715410 TI - Tribbles: novel regulators of cell function; evolutionary aspects. AB - Identification of rate-limiting steps or components of intracellular second messenger systems holds promise to effectively interfere with these pathways under pathological conditions. The emerging literature on a recently identified family of signalling regulator proteins, called tribbles gives interesting clues for how these proteins seem to link several 'independent' signal processing systems together. Via their unique way of action, tribbles co-ordinate the activation and suppression of the various interacting signalling pathways and therefore appear to be key in determining cell fate while responding to environmental challenges. This review summarises our current understanding of tribbles function and also provides an evolutionary perspective on the various tribbles genes. PMID- 16715412 TI - Anti-SARS drug screening by molecular docking. AB - Starting from a collection of 1386 druggable compounds obtained from the 3D pharmacophore search, we performed a similarity search to narrow down the scope of docking studies. The template molecule is KZ7088 (Chou et al., 2003, Biochem Biophys Res Commun 308: 148-151). The MDL MACCS keys were used to fingerprint the molecules. The Tanimoto coefficient is taken as the metric to compare fingerprints. If the similarity threshold was 0.8, a set of 50 unique hits and 103 conformers were retrieved as a result of similarity search. The AutoDock 3.011 was used to carry out molecular docking of 50 ligands to their macromolecular protein receptors. Three compounds, i.e., C(28)H(34)O(4)N(7)Cl, C(21)H(36)O(5)N(6), and C(21)H(36)O(5)N(6), were found that may be promising candidates for further investigation. The main feature shared by these three potential inhibitors as well as the information of the involved side chains of SARS Cov Mpro may provide useful insights for the development of potent inhibitors against SARS enzyme. PMID- 16715413 TI - Synthesis of alpha-trifluoromethyl alpha-amino acids with aromatic, heteroaromatic and ferrocenyl subunits in the side chain. AB - 5-Benzyloxy-4-trifluoromethyl-1,3-oxazoles, obtained from 5-fluoro-4 trifluoromethyloxazoles and benzyl alcohols, are capable for rearrangements. A 1,3 shift of a benzyl group is the key step of a new general route toward alpha trifluoromethyl substituted aromatic and heteroaromatic amino acids, demonstrating that 5-fluoro-4-trifluoromethyl-1,3-oxazole is a synthetic Tfm-Gly equivalent. On reaction with benzpinacol partially fluorinated oxazoles are transformed into bis(trifluoromethyl) substituted 2,5-diamino adipic acid and N benzoyl-2-benzhydryl-3,3,3-trifluoroalanine. PMID- 16715414 TI - Domino reactions with fluorinated five-membered heterocycles. alpha Trifluoromethyl alpha-amino acids with unsaturated side-chains. AB - Alpha-trifluoromethyl alpha-amino acids with unsaturated side-chains have been prepared from 5-fluoro-4-trifluoromethyloxazole and allyl, propargyl as well as terpene alcohols in a one-pot procedure. PMID- 16715415 TI - Striatal metabotropic glutamate receptors as a target for pharmacotherapy in Parkinson's disease. AB - Parkinson's disease (PD) is a common neurodegenerative disorder characterized by the loss of dopamine (DA)-containing neurons in the substantia nigra pars compacta (SNc). The symptoms are resting tremor, slowness of movement, rigidity and postural instability. Evidence that an imbalance between dopaminergic and cholinergic transmission takes place within the striatum led to the utilization of DA precursors, DA receptor agonists and anticholinergic drugs in the symptomatic therapy of PD. However, upon disease progression the therapy becomes less effective and debilitating effects such as dyskinesias and motor fluctuations appear. Hence, the need for the development of alternative therapeutic strategies has emerged. Several observations in different experimental models of PD suggest that blockade of excitatory amino acid transmission exerts antiparkinsonian effects. In particular, recent studies have focused on metabotropic glutamate receptors (mGluRs). Drugs acting on group I and II mGluRs have indeed been proven useful in ameliorating the parkinsonian symptoms in animal models of PD and therefore might represent promising therapeutic targets. This beneficial effect could be due to the reduction of both glutamatergic and cholinergic transmission. A novel target for drugs acting on mGluRs in PD therapy might be represented by striatal cholinergic interneurons. Indeed, the activation of mGluR2, highly expressed on this cell type, is able to reduce calcium-dependent plateau potentials by interfering with somato-dendritic N-type calcium channel activity, in turn reducing ACh release in the striatum. Similarly, the blockade of both group I mGluR subtypes reduces cholinergic interneuron excitability, and decreases striatal ACh release. Thus, targeting mGluRs located onto cholinergic interneurons might result in a beneficial pharmacological effect in the parkinsonian state. PMID- 16715416 TI - Characteristics of tumor extension requiring bronchoplasty and pneumonectomy in patients with lung cancer and positive bronchoscopic findings. AB - PURPOSE: The characteristics of tumor extension determine whether pneumonectomy or lobectomy with bronchoplasty should be performed for central lung cancer. We investigated how the characteristics of tumor extension determined the operative methods and the surgical outcomes. METHODS: We conducted a retrospective chart review of 151 patients with positive bronchoscopic findings who underwent lung cancer operations between January 1995 and March 2002. Twenty-five patients underwent pneumonectomy, 88 underwent lobectomy/segmentectomy (Lob/Seg), and 38 underwent Lob/Seg with bronchoplasty. RESULTS: Pathologic staging was higher in the pneumonectomy group than in the Lob/Seg groups, with or without bronchoplasty (P = 0.002). Interlobar extension and hilar lymph node involvement were more frequent, and mucosal invasion was less frequent, in the pneumonectomy group than in the Lob/Seg with bronchoplasty group. The frequencies of all specific pulmonary complications and 30-day mortality were similar among the three groups. The 5-year overall survival rates were 23.7%, 51.5%, and 72.8% for the pneumonectomy, Lob/Seg, and Lob/Seg with bronchoplasty groups, respectively (P = 0.0004). There was a significant difference in survival between patients with mucosal and those with submucosal types of lung cancer (P = 0.0114). CONCLUSIONS: Lob/Seg with bronchoplasty was feasible without a higher risk of operative complications or poorer long-term survival. The nature of tumor extension was important in the selection of operative methods and in predicting survival. PMID- 16715417 TI - Predictors of postoperative complications after radical resection for pulmonary aspergillosis. AB - OBJECTIVE: Although surgery has proven to be the best treatment for pulmonary aspergillosis, with the highest chance of achieving complete remission, it is difficult to determine the surgical indications for this disease because of the high incidence of postoperative complications. We conducted this study to identify some predictors of postoperative complications, in an attempt to reduce the morbidity rate. METHODS: We retrospectively analyzed the medical records of 31 patients (18 men, 13 women; median age 53 years) who underwent radical resection for pulmonary aspergillosis between 1976 and 2004. The clinical manifestations, surgical procedures, and postoperative complications were reviewed to clarify the predictors of postoperative complications. RESULTS: The morbidity rate associated with major complications such as intrapleural bleeding, bronchopleural fistula, and empyema, resulting in further surgery, was 19%. Univariate analysis revealed the predictors of major complications to be sex, severe preoperative symptoms, and extensive pulmonary resection. Multivariate analyses also indicated that preoperative symptom severity was an independent predictor of major complications. CONCLUSIONS: Pulmonary aspergillosis should be resected before the symptoms become too severe if the patient is a surgical candidate. PMID- 16715418 TI - Bacteriological evaluation of the cardiac surgery environment accompanying hospital relocation. AB - PURPOSE: Airborne bacteria in the environment are thought to be a cause of postoperative infection. With the relocation of our hospital, the operating room we had used for 35 years was replaced, changing the surgical environment for cardiac operation completely. We conducted this study to evaluate the bacteriological change in the surgical environment between the new and old operating rooms. METHODS: Airborne contaminants in the operating rooms were collected on blood agar plates, and samples of intraoperative salvaged blood from cardiac surgery were drawn from salvaged bags produced by Cell Saver 5 (Haemonetics, Braintree, MA, USA) in both the old (group O) and the new operating rooms (group N). These samples were cultured and evaluated bacteriologically. RESULTS: We collected nine samples of airborne contaminants from both group O and group N. The mean number of isolated bacteria colonies was 5.0 +/- 1.2 in group O, and 2.0 +/- 0.94 in group N (P < 0.001). Bacterial growth was detected in 85% of the salvaged blood samples from group O (n = 20) versus 60% from group N (n = 15) (P = 0.09). The mean bacteria count was 1.9 +/- 2.7 colony-forming units (cfu)/ml in group O versus 0.4 +/- 0.5 cfu/ml in group N (P = 0.032). CONCLUSION: Hospital relocation resulted in an improved operating room environment with less bacterial contamination of intraoperative salvaged blood. PMID- 16715419 TI - Efficacy of continuous low-dose human atrial natriuretic peptide given from the beginning of cardiopulmonary bypass for thoracic aortic surgery. AB - PURPOSE: Cardiac surgery performed under cardiopulmonary bypass (CPB) causes abnormalities of the renin-angiotensin-aldosterone system, resulting in decreased urine output and an accumulation of water in the third space. We studied the efficacy of continuous low-dose human atrial natriuretic peptide (hANP) in patients undergoing thoracic aortic surgery. METHODS: We divided 40 patients undergoing thoracic aortic surgery into two groups: the hANP group, which received 0.02 microg/kg per minute of hANP and the non-hANP group, which did not. The hemodynamics, urine output, intensive care unit (ICU) and hospital stay, bleeding volume, homologous blood transfusion volume, furosemide dose, corrected KCl volume, and postoperative respiratory, hepatic, and renal function were compared in the two groups. RESULTS: The urine output during CPB and from CPB weaning to return to ICU was significantly better in the hANP group. The bleeding volume, homologous blood transfusion volume, furosemide dose, and corrected KCl volume were all significantly less in the hANP group. CONCLUSIONS: These findings support the consensus that hANP exerts its diuretic effects to their full potential when administered continuously at low doses during thoracic aortic surgery. We found it to be effective for postoperative hemostasis and for preventing ischemic reperfusion injury. PMID- 16715420 TI - Receiver-operating characteristic analysis of leukocyte counts and serum C reactive protein levels in children with advanced appendicitis. AB - PURPOSE: To assess the diagnostic value of the white blood cell count (WBC) and serum C-reactive protein (CRP) level in children with advanced appendicitis. METHODS: We reviewed 289 children ranging in age from 3 to 15 years old, who underwent appendectomy in our hospital between 1993 and 2003. The advanced appendicitis group consisted of patients with associated perforation, abscess formation, or purulent ascites, and the simple appendicitis group consisted of patients who underwent uncomplicated appendectomy. We compared the age, sex, duration of abdominal pain before surgery, preoperative WBC and CRP level, and hospital stay in the two groups. We analyzed the diagnostic value of WBC and CRP level using a receiver-operating characteristic (ROC) curve. RESULTS: The children in the advanced appendicitis group had a significantly longer duration of pain, a higher serum CRP level, and longer hospitalization than those in the simple group. The serum CRP levels had good diagnostic value in differentiating between the advanced and simple groups. Both the CRP level and the WBC in the patients who suffered abdominal pain for longer than 48 h before surgery had diagnostic value in differentiating between the advanced and simple groups. CONCLUSION: These findings suggest that increased values of CRP and WBC more than 48 h after the onset of acute abdominal symptoms are indicators of advanced appendicitis. PMID- 16715421 TI - Liver injury in sepsis and abdominal compartment syndrome in rats. AB - PURPOSE: To evaluate the extent of liver injury after the onset of sepsis and abdominal compartment syndrome (ACS) in rats. METHODS: We divided 60 rats into four groups of 15. Group 1 was the sham group. In group 2, sepsis was induced by cecal puncture and ligation; in group 3, ACS was created by placing a catheter in the abdominal cavity; and in group 4, both sepsis and ACS were induced simultaneously. Liver sections stained with hematoxylin-eosin were assessed pathologically, and liver injury was defined by the following five pathological patterns: spotty necrosis, capsular inflammation, portal inflammation, ballooning degeneration, and steatosis of the liver. We revised a new scoring system, called "Hepatic Injury Severity Scoring" (HISS), to evaluate the liver injury in sepsis, ACS, and sepsis plus ACS. Blood was collected for liver function tests. RESULTS: The total scores of groups 1, 2, 3, and 4 were 18, 92, 86, and 123, respectively. There were significant differences in histopathologic grade between group 1 and groups 2, 3, and 4 (P < 0.05). Aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and total bilirubin levels were significantly higher in group 4 than in the other three groups. CONCLUSIONS: The findings of this study showed that liver function severely affected the onset of ACS and sepsis. The liver injury resulting from sepsis plus ACS is more severe than that resulting from either one independently. PMID- 16715422 TI - Isolated unilateral absence of a pulmonary artery treated by pneumonectomy in an adult: report of a case. AB - Congenital unilateral absence of a pulmonary artery (UAPA) is a rare anomaly usually diagnosed in childhood. We report a case of isolated UAPA in an adult without any other cardiovascular anomaly. The patient was admitted for repeated progressive hemoptysis, which we treated with embolization of the bronchial artery three times, despite which the hemoptysis kept recurring. Finally, the patient was treated successfully by right pneumonectomy. Thus, we think that surgical intervention is indicated for recurrent hemoptysis in patients with isolated UAPA. PMID- 16715423 TI - Massive pulmonary embolism complicated by a patent foramen ovale with straddling thrombus: report of a case. AB - We report a case of massive right pulmonary embolism with a patent foramen ovale and straddling thrombus, occurring a few days after cesarean section in a 31-year old woman. Preoperatively, a mass was seen echocardiographically in four cardiac cavities. We performed emergency surgery because of the patient's acute hemodynamic deterioration. Intraoperatively, we found a thrombus entrapped in the patent foramen ovale. Most of the thrombus was floating in the right atrium and a long end was found in the left atrium, in addition to the pulmonary emboli. We removed the thrombus, closed the patent foramen ovale by direct suturing, and performed pulmonary embolectomy. Histological examination confirmed thrombi. Doppler examination of the venous system did not reveal any possible source of the thrombus. The patient is now well and free from recurrence of embolic disease 1 year after surgery. We review the literature on this relatively unusual thromboembolic disease. PMID- 16715426 TI - Proximal aortic replacement with ascending-descending bypass for a diffuse aneurysm: report of a case. AB - We performed successful ascending-arch aortic replacement and concomitant ascending-to-descending aorta bypass with exclusion of a descending thoracic aneurysm, via median sternotomy, for a ruptured aortic aneurysm involving the entire thoracic aorta. The patient was an 80-year-old man with cardiopulmonary dysfunction and a history of lung tuberculosis. This operation, which has been used for complex descending thoracic aortic lesions such as recoarctation, is a feasible option for a diffuse thoracic aortic aneurysm when single-stage repair is mandatory. PMID- 16715425 TI - Coronary artery bypass grafting under cardiopulmonary bypass in a patient with beta-thalassemia: report of a case. AB - We performed successful coronary artery bypass grafting under cardiopulmonary bypass (CPB) in a man with beta-thalassemia. Cardiopulmonary bypass for patients with hemoglobin disorders occasionally results in fatal hematologic complications. However, taking careful precautions such as transfusing stored autologous blood and administering haptoglobin may eliminate the need for allotransfusion, and prevent severe or persistent hemolysis and other complications. This case shows that cardiac surgery can be performed safely under CPB in patients with beta-thalassemia. PMID- 16715424 TI - Excision after chemoradiotherapy of invasive thymoma extending into the right atrium: report of a case. AB - We report a case of invasive thymoma with intracardiac extension, resulting from the progression of intracaval growth, in a 56-year-old woman. Initially, the patient received two courses of chemotherapy, but the tumor showed only a modest response; however, subsequent radiotherapy reduced the tumor size further and the intracardiac lesion disappeared, making it possible to excise the tumor without cardiopulmonary bypass. Thus, when a thymoma does not respond well, we recommend radiotherapy as another treatment option, because its effects may allow for less invasive and more complete tumor excision. PMID- 16715427 TI - Aortoenteric fistula after endovascular stent grafting for an abdominal aortic aneurysm: report of a case. AB - We report a case of an aortoenteric fistula (AEF) developing after endovascular stent grafting (EVSG) for an abdominal aortic aneurysm (AAA). A 69-year-old male patient with a history of panperitonitis caused by rectal perforation underwent EVSG for an AAA. A follow-up contrast-enhanced computed tomography (CT) scan, done 12 months after the EVSG, confirmed shrinkage of the AAA with no endoleak. However, 19 months postoperatively, an AEF developed between the AAA and the jejunum. Although there was no endoleak on a subsequent CT scan, we noted enlargement of the AAA and inflammatory changes in the surrounding tissue. The patient was treated surgically and discharged in good health 74 days postoperatively. Thus, one should consider the possibility of this devastating complication, even in patients without an endoleak, after EVSG for AAA. PMID- 16715428 TI - Successful management of esophageal perforation diagnosed 3 days after injury caused by an explosion in the workplace: report of a case. AB - We report a case of esophageal perforation caused by an explosion, but which was not diagnosed until 3 days after the injury. A 53-year-old worker sustained superficial dermal burns to his trachea, face, neck, and legs during an explosion. The burns were treated conservatively at a local hospital, but he was transferred to our hospital 3 days after the injury, when mediastinal emphysema and bilateral pleural effusion became evident. An esophagogram followed by computed tomography showed an esophageal perforation caused by the blast injury, and we performed an esophagectomy with recontruction of the gastric tube. After the operation, an X-ray showed a foreign body in the lower abdomen, which we found in the upper thoracic esophagus on the day of injury. We surmised that the patient had inadvertently swallowed a foreign body, which had been heated and scattered by the explosion, and it had melted the upper thoracic esophagus. PMID- 16715429 TI - Long-term survival after resection of metachronous bilateral adrenal metastases of mucinous gastric carcinoma: report of a case. AB - We report a case of metachronous bilateral adrenal metastases from mucinous adenocarcinoma of the stomach. A 68-year-old man who had undergone surgery for advanced gastric cancer 5 months earlier had a follow-up computed tomography (CT) scan, which showed a right adrenal tumor. We performed a right adrenalectomy, and histopathological examination revealed a mucinous adenocarcinoma with features consistent with those of gastric cancer. A routine follow-up CT scan done 41 months after the right adrenalectomy showed a left adrenal mass. Chemotherapy had no apparent effect, and left adrenalectomy was performed 65 months after the right adrenalectomy. Histopathological examination also revealed a metastasis from gastric cancer. The patient was alive without recurrence 40 months after the left adrenalectomy. This case suggests that resection of adrenal metastasis from gastric cancer is an effective treatment option that may prolong survival in selected patients. PMID- 16715430 TI - Intrahepatic cholangiocarcinoma with lymph node metastasis successfully treated by immunotherapy with CD3-activated T cells and dendritic cells after surgery: report of a case. AB - Intrahepatic cholangiocarcinoma (ICC) with lymph node (LN) metastasis is generally associated with a poor prognosis. However, we treated ICC with LN metastasis successfully by surgery and postoperative immunotherapy in a 59-year old woman. The immunotherapy consisted of CD3-activated T cells and tumor lysate- or peptide-pulsed dendritic cells. Pathological examination confirmed a diagnosis of moderately differentiated adenocarcinoma with LN metastasis and portal vein invasion. The patient has been alive without recurrence for 3 years 6 months since her operation. PMID- 16715431 TI - Intramural hematoma of the cecum as the lead point of intussusception in an elderly patient with hemophilia A: report of a case. AB - Hemophilia A is a congenital bleeding disorder characterized by a deficiency of coagulation factor VIII. Intramural hematoma of the colon is a very rare complication of this disease. We report a case of intramural hematoma of the cecum serving as the lead point of intussusception in a 65-year-old man with hemophilia A. The patient presented with right-sided abdominal pain and bloody stool. Palpation of his abdomen revealed a fist-sized mass. Abdominal computed tomography (CT) showed a circular mass with concentric rings, consistent with an intussuscepted intestine. Because his activated partial thromboplastin time (APTT) was prolonged, we gave him a continuous infusion of factor VIII during and after surgery. Laparotomy revealed an irreducible colo-colic intussusception and we identified a cecal hematoma as the lead point. After an unsuccessful attempt at Hutchinson's maneuver, we performed right colectomy. We report this case to illustrate the necessity of monitoring APTT in patients with hemophilia A who undergo surgery. PMID- 16715432 TI - Immediate postoperative angiographic embolization after damage control surgery for liver injury: report of a case. AB - A multimodality strategy, including damage control and angioembolization techniques, has been reported to reduce the mortality associated with surgery for complex blunt hepatic injuries. However, the indications for angiographic evaluation and embolization in patients who require surgery for hepatic injury remain unclear. We report a case of blunt hepatic injury requiring emergency laparotomy, which we treated by damage control surgery because of an inaccessible major venous injury and the fact that coagulopathy was stopping hemostasis. The decision to perform immediate postoperative angiography was based on the hemorrhagic response to Pringle's maneuver and its release after perihepatic packing during surgery. Hepatic angiography revealed extravasation from a branch of the middle hepatic artery, which was embolized successfully. Although the definitive indications for immediate postoperative angioembolization for hepatic injury have not been established, the hemorrhagic response to Pringle's maneuver and its release after perihepatic packing during damage control surgery is an indication for immediate postoperative angioembolization. PMID- 16715433 TI - Posterior esophagopexy with dome formation to prevent reflux in interposed jejunal pouch reconstruction after total gastrectomy. AB - Jejunal pouch interposition (JPI) reconstruction after total gastrectomy has proven effective for improving postoperative quality of life; however, evaluation of bile reflux into the esophagus shows that the reflux of digestive juice is not sufficiently prevented. Therefore, in addition to the conventional reconstruction technique, we created an artificial pouch to prevent the reflux of digestive juice from the jejunal pouch into the esophagus, and performed a new surgical technique based on the Hill's posterior gastropexy. No postoperative complications were observed and the postoperative measurement showed a decrease in the duration of bile reflux into the esophagus. Thus, our new surgical procedure seems to effectively prevent bile reflux. PMID- 16715434 TI - Plastron-trapdoor technique for direct-vision sternal re-entry. AB - Repeat median sternotomy does not allow good access for retrosternal re-entry. Conversely, by using the plastron-trapdoor technique, which we describe in this report, the retrosternal tissues are dissected from the body of the sternum under direct vision, thus allowing relatively safe and wide exposure for retrosternal re-entry. PMID- 16715435 TI - Sentinel Lymph Node Biopsy for Cutaneous Head and Neck Melanoma: Mapping the Parotid Gland. AB - BACKGROUND: Sentinel lymph node biopsy (SLNB) for primary cutaneous head and neck melanoma (CHNM) has been shown to be successful and is the current standard of care for intermediate-thickness melanoma. We evaluated our experience with CHNM associated with SLNB mapping to the region of the parotid gland. METHODS: Retrospective review of a prospectively collected melanoma database identified 1014 CHNMs. Two-hundred twenty-three patients underwent SLNB, and 72 (32%) had mapping in the region of the parotid gland between May 1995 and June 2003. RESULTS: The mean number of SLNs per patient was 2.5. A sentinel lymph node (SLN) was successfully identified in 94% of patients, and in 12%, the SLN was positive for metastatic disease. Biopsy of intraparotid SLNs was performed in 51.4% and of periparotid SLNs in 26.4%, and a superficial parotidectomy was performed in 22.2%. Ten patients were found to have lymph nodes in the parotid region with metastatic disease (eight identified by SLNB), and two (20%) patients developed intraparotid lymph node recurrence in the setting of a negative SLNB. Same-basin recurrence in SLN-negative patients was 3.3% with a median follow-up of 26 months. Facial nerve dysfunction was identified in seven (10%) patients. Facial nerve function returned to preoperative status in all patients. CONCLUSIONS: SLNB for patients with primary CHNM mapping to the parotid gland can be performed with a high degree of accuracy and a low morbidity consisting of temporary facial nerve paresis. PMID- 16715450 TI - Lack of association between microsatellite instability and benign adrenal tumors. AB - BACKGROUND: The adrenal gland may give rise to pheochromocytomas, which are catecholamine-producing tumors originating from the adrenal medulla, or to adrenocortical tumors, which derive from the adrenocortical cortex and may be secreting or not. The genetic mechanisms underlying the formation of these tumors include somatic mutations in susceptibility genes, especially in the familial forms, and allelic loss, especially in chromosome 1. AIM: The aim of this study was to investigate a third genetic mechanism by evaluating microsatellite instability using the reference markers (Bat25, Bat26, D2S123, D5S346, D17S250) validated by the National Cancer Institute. Microsatellite loci were analyzed in 32 benign tumors, including 11 pheochromocytomas and 21 adrenocortical tumors, in patients with and without familial syndrome. RESULTS: The different alleles of microsatellite loci were reliably detected by DNA fragments analysis, whereas data obtained after melting-point analysis on the Lightcycler were inconsistent. No microsatellite instability was detected in any tumor. One patient with a unilateral pheochromocytoma showed a loss of heterozygosity for D17S250. A second patient with a MEN-2A syndrome and a two-sided pheochromocytoma exhibited a loss of heterozygosity for D2S123 in the right tumor only and a retention of heterozygosity for all markers in the left tumor. CONCLUSIONS: These results suggest that microsatellite instability, evaluated by the five reference markers of the National Cancer Institute, is not a feature of benign adrenal tumors. PMID- 16715451 TI - The role of whole-body fluorine-18-FDG positron emission tomography in the detection of recurrence in symptomatic patients with stages II and III breast cancer. AB - BACKGROUND: The role of whole-body fluorine-18-FDG positron emission tomography (FDG-PET) as an adjunct localize recurrence in stages II and III breast cancer patients who present with clinical suspicion for recurrence is not well established. We report our experience in such a patient population. METHODS: A retrospective review of all patients with stages II and III breast cancer who had a whole-body FDG-PET scan was performed. RESULTS: Of the 23 patients who fit the criteria, 9 had stage II and 14 had stage III breast cancer. Overall sensitivity, specificity, and accuracy were 81%, 100%, and 87%, respectively. Positive and negative predictive values for stages II and III were 100% and 83%, respectively, and 100% and 50%, respectively. FDG-PET detected two recurrences that were missed by conventional imagings, but such recurrences were local and amenable for biopsy. CONCLUSIONS: In patients with stages II and III breast cancer who present with a suspicion for recurrent disease, a whole-body FDG-PET scan may be a useful adjunct in the evaluation of recurrence. However, its added benefit over conventional imaging should be questioned. PMID- 16715452 TI - Respiratory function after laparoscopic distal gastrectomy--an index of minimally invasive surgery. AB - BACKGROUND: As the techniques of laparoscopic surgery have improved, various institutions have performed laparoscopic gastrectomies with regional lymph node dissection, as well as open surgery. Although alleviation of postoperative pain and prompt recovery have been reported in the literature, objective indexes of the minimal invasiveness of laparoscopic procedures are as yet very few. METHODS: We performed distal gastrectomy with regional lymph node dissection for gastric cancer patients using three kinds of procedures, namely, open gastrectomy, hand assisted laparoscopic surgery (HALS), and totally laparoscopic gastrectomy. Ablation of the stomach, lymph node dissection, and reconstruction of the digestive tract were all carried out intracorporeally with or without HALS in the laparoscopic procedures. The ordinary respiratory function test was performed pre and postoperatively for 50 patients operated on by each procedure, and the reduced percentages of the measured values were calculated. RESULTS: Postoperative respiratory function was consistently excellent, with minimal loss of vital capacity and forced expiratory volume per second in the totally laparoscopic group compared to HALS or open cases. CONCLUSIONS: Although it may be a complicated technique, totally laparoscopic distal gastrectomy is considered a minimally invasive procedure for gastric cancer from the viewpoint of postoperative respiratory function. PMID- 16715453 TI - Hot stuff--can't get enough. PMID- 16715456 TI - Disseminated herpes virus (HSV-2) infection with rhabdomyolysis and hemophagocytic lymphohistiocytosis in a patient with bone marrow failure syndrome. PMID- 16715455 TI - Differential diagnosis of solitary pulmonary nodules based on 99mTc-EDDA/HYNIC TOC scintigraphy: the effect of tumour size on the optimal method of image assessment. AB - PURPOSE: The aim of the study was to determine an optimal method for the evaluation of scintigrams obtained with (99m)Tc-EDDA/HYNIC-TOC for the purpose of differential diagnosis of solitary pulmonary nodules (SPNs) and to assess the diagnostic value of the method. METHODS: Eighty-five patients (48 males and 37 females, mean age 57 years, range 34-78 years) were enrolled in the study. Patients underwent (99m)Tc-EDDA/HYNIC-TOC scintigraphy for the purpose of differential diagnosis of SPNs (size between 1 and 4 cm). Images of all patients were evaluated visually in a prospective manner. RESULTS: Positive scintigraphic results were found in 37 out of 40 (93%) patients with malignant SPNs including 34 out of 35 (97%) patients with primary lung carcinoma. Two remaining false negative cases turned out to be metastatic lesions of malignant melanoma and leiomyosarcoma. Among 45 benign tumours, negative results were obtained in 31 cases (69%) and positive results in 14. The accuracy of the method was 80%. Analysis of the results of the visual assessment of scintigrams revealed a significantly higher frequency of false positive results among larger nodules (diameter at least 1.4 cm). Uptake of the tracer in those nodules was therefore assessed semi-quantitatively (using the tumour-to-background ratio), in expectation of an improvement in the low specificity of the visual method. The semi-quantitative assessment reduced the total number of false positive results in a subgroup of larger nodules from 13 to six, while preserving the high sensitivity of the method. CONCLUSION: The combination of visual analysis (for lesions smaller than 1.4 cm in diameter) and semi-quantitative assessment (for larger lesions) provided a high sensitivity of the method and significantly improved its specificity (84%) and accuracy (88%) in comparison with visual analysis (p<0.05). PMID- 16715457 TI - Labrum repair combined with arthroscopic reduction of capsular volume in shoulder instability. AB - We performed arthroscopic treatment of traumatic anterior and anteroinferior shoulder instability combining three procedures--labrum repair, reduction of capsular volume and suture of the rotator cuff interval--with the aim of analysing the results with regard to stability and function. Between January 1999 and December 2003, 27 patients underwent arthroscopic treatment for labrum repair with metal anchors, reduction of capsular volume through thermal capsulorrhaphy and suture of rotator cuff interval. These patients were evaluated in the pre- and postoperative period using the UCLA and Rowe scales and in the postoperative period using the ASES scale. During a mean follow-up period of 32.4 months (range 22-74 months) all shoulders remained stable. Using the UCLA scale, there was improvement from the preoperative period, with a mean score of 24.7, to the postoperative period, with a mean of 32.81. Improvement was also shown by the Rowe scale, with a mean score of 39.81 in the preoperative period and 90.74 in the postoperative period. On the ASES scale the mean score was 92.22. All shoulders remained stable and there was marked functional improvement in the patients who were treated. These results are comparable to those obtained with open surgery, observing similar patient selection criteria. PMID- 16715458 TI - [The Resilience Scale (RS): a statistical comparison of the short and long version based on a patient population]. AB - The use of short and reliable questionnaires in psychological and medical studies is increasingly demanded due to economic, but also ethical reasons. The expectation of low dropout rates within longitudinal studies is an important issue with respect to the validity and quality of clinical studies. In this article, differences of the Resilience Scale (RS) in a short (11 items) and a long version (25 items) are described related to 212 patients treated in a radiooncological unit before receiving radiotherapy. It can be demonstrated that the long and the short version of the Resilience Scale did not appear to differ clinically relevant (differences of means = - 0.080, median = - 0.042, effect size = 0.11). Effect sizes and correlations of the resilience of the patients and other illness- and treatment-related variables such as the need for psychosocial support and fatigue did not change depending on the use of the short or long version of the questionnaire. Therefore, it can be concluded that the short version of the Resilience Scale turns out to be a useful and valid instrument that can be recommended for its use in psychological and medical research. PMID- 16715460 TI - [Mental health impact of stalking in men and women]. AB - Using data from the Mannheim stalking study, the present report analyses gender differences with regard to various mental health indicators and potential mediator effects of stalking victimization. Furthermore, we were interested in whether the impact of stalking on mental health was comparable for men and women. The study included a postal survey of 675 community residents on the experience of intruding harassment and on mental health indicators. In the Patient Health Questionnaire (PHQ-D, Lowe et al. 2001), women scored higher on most of the subscales (depression, anxiety, somatic complaints, stress, psychosocial impairment) than men. Furthermore, more women fulfilled criteria for at least one threshold or subthreshold mental disorder syndrome according to DSM-IV, and more women were under psychotropic medication. However, the identified associations were completely mediated by the higher prevalence of stalking victims in women. In contrast, the associations of stalking victimization with poor mental health, psychosocial functioning, and use of medication were largely comparable across gender. PMID- 16715459 TI - [The distressed personality (type D)--correlations with anger, aggression and hostility]. AB - Type D (distressed personality) comprises the joint tendency to experience negative emotions (negative affectivity) and to inhibit these emotions in social situations (social inhibition). The distressed personality can be assessed by the Type D scale (DS14). In the present study, the German version of the DS14 was to be validated especially with regard to its interrelation with anger, aggression and hostility. The study included a clinical sample of 90 patients with coronary heart disease and a nonclinical sample of 86 participants. The two-factor structure of the DS14 could be confirmed in both samples. As expected, the subscale negative affectivity correlates highly with neuroticism; the subscale social inhibition correlates highly with extraversion. Negative affectivity also relates to almost every anger and aggression scale applied in this study. Participants classified as Type D report higher anger experience than non-Type D classified persons. They show both heightened anger-in and anger-out and encounter others with more cynicism and hostility. It seems promising to provide special psychological treatment for these patients. PMID- 16715461 TI - [Validation of the German version of the Inflammatory Bowel Disease Questionnaire (Competence Network IBD, IBDQ-D)]. AB - The Inflammatory Bowel Disease Questionnaire (IBDQ) is the standard disease specific instrument for assessment of health-related quality of life (HRQOL) in patients with inflammatory bowel diseases (IBD). A German translation has not been validated. 415 outpatient IBD-patients (Crohn's Disease n = 306, Ulcerative Colitis n = 109) completed the German version of the IBDQ (Competence network IBD, IBDQ-D), the Hospital Anxiety and Depression Scale German Version (HADS-D) and the Questions on Life Satisfaction FLZ. Face validity was assessed by a physicians' and patients' panel. Disease activity was measured by the German Inflammatory Bowel Disease Activity Index (GIBDI). With 97.3 % completed items the acceptance was high. The Cronbach's alpha for the subscales ranged from 0.88 to 0.89. The correlation coefficients with comparable subscales of other instruments ranged between 0.09 and 0.70. Patients in remission and different disease activities differed significantly (p < 0.001) in all IBDQ-D-subscales. PMID- 16715462 TI - [Psychosomatic care practice and cooperation needs as seen by physicians and psychologists providing outpatient care]. AB - Cooperation in psychosocial care is gaining increasing importance due to newly established psychosomatic departments in the acute care setting and to structural changes following the Health Care Modernization Act. The presented study aims at describing the psychosomatic treatment and cooperation practice in the ambulatory sector and the cooperation possibilities with the inpatient psychosomatic care sector. Registered doctors in all specialties along with psychotherapists working in private practice in the southwestern region of the Association of Panel Doctors (Nordbaden) were surveyed about this in a written questionnaire in 2003. Respondents totaled 497 (33 %). General practitioners (GPs) and somatic specialists first estimated the psychosocial treatment necessities based on patients in their own practice. Half reported cooperation with a psychosomatic colleague, 10 % took part in a Balint-style group. GPs and somatic specialists unlicensed in psychosomatic basic care reported significantly lower numbers on the existing psychosocial care practice compared to those with the respective license. GPs and somatic specialists regard diagnostic procedures as central for improving their daily psychosomatic/psychosocial care. More than 50 % would request a brief diagnostic screening instrument. Psychiatrists and psychotherapists prefer the option of case conferences. The reported cooperation preferences of a psychosomatic department follow established setting structures with little mention of cooperation options falling outside established sectors. GPs and psychiatrists/psychotherapists share the same priority in designating a contact person in the department for crisis intervention. This expresses their feelings for greater need in direct and acute cooperation features. Somatic specialists prefer to request a second opinion from the department. The predominant diagnoses to admit patients for both psychiatrists and psychotherapists are eating disorders, pain syndromes, and personality disorders. PMID- 16715463 TI - [Lay theories regarding myocardial infarction in a transcultural comparison]. AB - Culturally influenced lay theories about myocardial infarction which exist in healthy individuals have an impact on treatment compliance. However, empirical data on the subject is rare. Using healthy subjects, a transcultural survey comparing three different ethnic groups was conducted. The groups were: Germans in Germany, Spaniards in Spain, and 1st generation Spaniards in Germany. Subjects were paralleled according to age, sex, and education. The groups were compared regarding cultural differences in casual attributions and locus of control with respect to myocardial infarction. While all three groups show a psycho-social understanding of myocardial infarction, it is most predominate in the German group. The results show both common factors as well as some significant differences between Germans and Spaniards, the Spaniards reporting more external attributions. Consequences for prevention concepts and medical care in a multicultural society were derived from the results. PMID- 16715464 TI - Evaluation of the internal temperatures of an 8.6 kDa protein cation exposed to a hot dispenser cathode employed in electron capture dissociation mass spectrometry. AB - The 'effective' internal temperature of an 8.6 kDa ubiquitin cation was estimated under electron capture dissociation (ECD) conditions, in which a dispenser cathode electron source was mounted just outside an ion cyclotron resonance (ICR) cell, i.e., axially displaced at a distance less than 1 cm from the rear trap plate of the ICR cell. In this ECD configuration, thermal activation of the molecular ions stored in the ICR cell was anticipated since the heated dispenser cathode (T(cathode surface) > 1000 degrees C) emitted a large amount of (both visible and infrared) radiation as well as electrons. An evaluation of the internal temperature of ubiquitin 6+ and 7+ cations was made by comparing our ECD fragmentation patterns with those obtained by McLafferty et al. (J. Am. Chem. Soc. 2002; 124: 6407) as a function of the ion temperature. In McLafferty's configuration, the heating (or thermal activation) effect of their filament source was minimal since the filament was displaced by a distance as far as 70 cm from their ICR cell. A careful comparison reveals that the fragmentation patterns obtained in this work are very similar to those previously measured at T approximately 125 degrees C. In terms of sequence coverage, our ECD configuration provides better results, and in particular without the aid of any other simultaneous activation method, such as thermal heating, infrared multiphoton irradiation, or collisional activation, except for the visible and infrared radiation from the heated cathode. PMID- 16715465 TI - Fetal arrhythmias. PMID- 16715466 TI - New models to predict depth of infiltration in endometrial carcinoma based on transvaginal sonography. AB - OBJECTIVES: Preoperative knowledge of the depth of myometrial infiltration is important in patients with endometrial carcinoma. This study aimed at assessing the value of histopathological parameters obtained from an endometrial biopsy (Pipelle de Cornier; results available preoperatively) and ultrasound measurements obtained after transvaginal sonography with color Doppler imaging in the preoperative prediction of the depth of myometrial invasion, as determined by the final histopathological examination of the hysterectomy specimen (the gold standard). METHODS: We first collected ultrasound and histopathological data from 97 consecutive women with endometrial carcinoma and divided them into two groups according to surgical stage (Stages Ia and Ib vs. Stages Ic and higher). The areas (AUC) under the receiver-operating characteristics curves of the subjective assessment of depth of invasion by an experienced gynecologist and of the individual ultrasound parameters were calculated. Subsequently, we used these variables to train a logistic regression model and least squares support vector machines (LS-SVM) with linear and RBF (radial basis function) kernels. Finally, these models were validated prospectively on data from 76 new patients in order to make a preoperative prediction of the depth of invasion. RESULTS: Of all ultrasound parameters, the ratio of the endometrial and uterine volumes had the largest AUC (78%), while that of the subjective assessment was 79%. The AUCs of the blood flow indices were low (range, 51-64%). Stepwise logistic regression selected the degree of differentiation, the number of fibroids, the endometrial thickness and the volume of the tumor. Compared with the AUC of the subjective assessment (72%), prospective evaluation of the mathematical models resulted in a higher AUC for the LS-SVM model with an RBF kernel (77%), but this difference was not significant. CONCLUSIONS: Single morphological parameters do not improve the predictive power when compared with the subjective assessment of depth of myometrial invasion of endometrial cancer, and blood flow indices do not contribute to the prediction of stage. In this study an LS-SVM model with an RBF kernel gave the best prediction; while this might be more reliable than subjective assessment, confirmation by larger prospective studies is required. PMID- 16715467 TI - Support vector machines versus logistic regression: improving prospective performance in clinical decision-making. PMID- 16715468 TI - Precursor ion scanning for the non-targeted detection of individual arsenosugars in extracts of marine organisms. AB - Arsenosugars are a group of arsenic compounds reported to be present in a wide variety of marine organisms. Numerous such compounds have been identified and characterized in marine organisms; however, unknown arsenosugar species may also be present. This indicates the need for an analytical technique suitable for their non-targeted detection. One such technique is tandem mass spectrometry operated in the precursor ion scanning mode. This technique is based on scanning for precursor ions that give specific product ions, characteristic of the compounds under investigation. In the present study two subgroups of arsenosugar species were examined, the oxo- and the thioarsenosugars, the CID behavior of which is well known from previous studies. In the case of the oxoarsenosugars characteristic product ions were observed at m/z 237 and 97, and for the thioarsenosugars at m/z 253 and 97. Validation of this approach was carried out by analyzing extracts of two commercial kelp powders with known contents of arsenosugar species. All arsenosugars reported to exist in these materials were detected successfully using the precursor ion scanning approach. The limits of detection for the oxo- and the thioarsenosugar species, and the selectivity and sensitivity of the method, strongly indicate the suitability of this approach for the non-targeted detection of arsenosugars in extracts of marine origin. PMID- 16715469 TI - Determination of phenylalanine in human serum by isotope dilution liquid chromatography/tandem mass spectrometry. AB - Isotope dilution liquid chromatography/tandem mass spectrometry (ID-LC/MS/MS) has been developed as a candidate reference method to determine the level of phenylalanine in human serum. The advantages of this method include a simple sample preparation without derivatization, selective detection of analytes, and the use of an isotopic analogue as an internal standard. Phenylalanine and its isotopic analogue, phenylalanine-ring-(13)C(6), were monitored at the transitions m/z 166.2/120.2 and 172.2/126.2 in the multiple-reaction monitoring (MRM) mode, respectively. The expanded uncertainty of the measurement result of phenylalanine in the serum was approximately 1.2% within a 95% confidence level. A standard reference material, with a certified value of phenylalanine, was analyzed in order to verify this method. The result obtained by the ID-LC/MS/MS method differed somewhat from the certified value, but agreed well with the gravimetric value. The measurement result of phenylalanine in serum by ID-LC/MS/MS was compared with the results from the commercial HPLC method, which was carried out in clinics. The results from the commercial HPLC method showed inconsistent results with each other. The busted results from the commercial HPLC method suggest that it should be possible to trace the results of the commercial fields to well-characterized reference materials or methods. PMID- 16715470 TI - Liquid chromatography/tandem triple-quadrupole mass spectrometry for determination of paclitaxel in rat tissues. AB - A liquid chromatography/tandem triple-quadrupole mass spectrometry assay to quantify paclitaxel in rat tissue homogenates containing taxol or paclitaxel nanoliposome (PTX-NLP) was developed and validated. Liquid-liquid extraction with tert-butyl methyl ether was used for tissue sample preparation and docetaxel was used as the internal standard. Paclitaxel and docetaxel were separated on a 200 mm x 4.6 mm x 5 microm C(18) column and quantified using a triple-quadrupole mass spectrometer operating in positive ion electrospray selective reaction monitoring mode (ESI(+)-SRM) with a total run time of 6.0 min. The peak area of the m/z 876.3 --> 307.9 transition of paclitaxel is measured versus that of the m/z 830.3 --> 549.1 transition of docetaxel to generate the standard curves. The standard curves were linear over the concentration range of 0.2008-2008 ng/mL for different tissues. The method had high extraction recovery (>90%) and accuracy (>90%) with the intra-day and inter-day precision <15%. Frozen stability, freeze/thaw stability, extraction stability and solution stability at ambient temperature were examined, which indicated the tissue samples should be extracted within 5 days and avoid being frozen and thawed repeatedly over 5 times. Extracted samples after evaporation could be stored at -20 degrees C for 20 days without drug degradation and no degradation was also observed after solution samples were left to stand at ambient temperature for 24 h. This assay was used to support an in vivo biodistribution study of PTX-NLP in rats. PMID- 16715471 TI - Reactions of platinum cluster ions with benzene. AB - In this work, the cation and anion products of the reactions between platinum clusters produced by laser ablation and the benzene molecules seeded in argon have been studied using a high-resolution reflectron time-of-flight mass spectrometer (RTOFMS). The dominant cation products are [C(6n)H(6n - k)](+) and [Pt(m)(C(6)H(6))(n)](+) complexes, while the dominant anion products are dehydrogenated species, [C(6)H(5)PtH](-), [PtC(12)H(k)](-) and [Pt(m)C(6)H(4) . . . (C(6)H(6))(n)](-), etc. Some important intermediate structures ([PtC(6)H(6)](+), [Pt(C(6)H(6))(2)](+), [Pt(2)(C(6)H(6))(3)](+), [C(6)H(5)PtH]( ), [Pt(2)C(6)H(4)](-), [Pt(3)C(6)H(4)](-) and [Pt(4)C(6)H(4)](-)) have been analyzed using density functional theory (DFT) calculations. Different reaction mechanisms are proposed for platinum cluster cations and anions with benzene, respectively. PMID- 16715472 TI - Top-down analysis of basic proteins by microchip capillary electrophoresis mass spectrometry. AB - A system of microchip capillary electrophoresis/electrospray ionization mass spectrometry (microchip-CE/ESI-MS) for rapid characterization of proteins has been developed. Capillary electrophoresis (CE) enables rapid analysis of a sample present in very small quantity, such as at femtomole levels, at high resolution. Faster CE/MS analysis is expected by downsizing the normal capillary to the microchip (microchip) capillary. Although rapidity and high resolution are advantages of CE separation, electroosmotic flow (EOF) instability caused by the interaction between proteins and the microchannel surface results in low reproducibility in the analysis of basic proteins under neutral pH conditions. By coating the microchannel surface with a basic polymer, polyE-323, basic proteins, which have pI values of over 7.5, could be separated and detected by microchip CE/MS on quadrupole (Q) and time-of-flight (TOF) hybrid instruments. By increasing the cone and collision voltages during the analysis by microchip CE/ESI-MS of a small protein, some product ions, which contain the sequence information, could also be obtained, i.e., 'top-down' analysis of the protein could be accomplished with this microchip-CE/MS system. To our knowledge, this is the first report of 'top-down' analysis of a protein by microchip-CE/MS. Since it requires a much shorter time and a smaller sample amount for analysis than the conventional liquid chromatography (LC)/ESI-MS method, microchip-CE/MS promises to be suitable for the high-throughput characterization of proteins. PMID- 16715473 TI - Apoptosis resistance and response to chemotherapy in primary nodal diffuse large B-cell lymphoma. AB - Diffuse large B-cell lymphomas (DLBCL) represent the most common type of adult malignant lymphoma in western countries and are treated with high dose combination chemotherapy. Although initially the majority of patients respond to this therapy, many do not achieve complete remission and others experience an early relapse. Several studies have shown that prediction of the clinical response to chemotherapy is possible before the start of chemotherapy treatment. Apparently, DLBCL are intrinsically either resistant or sensitive to chemotherapy induced cell death. Differences in functional integrity of the apoptosis cascade are an important factor predicting outcome in DLBCL. In this review we discuss the possible mechanisms leading to intrinsic resistance to apoptosis and provide an explanation why strong differences in apoptosis sensitivity between DLBCL are observed. Subsequently we will focus on how differences in this intrinsic apoptosis resistance provide an explanation for the variable response to combination chemotherapy and how this can be used for further therapy tailoring. PMID- 16715474 TI - Analysis of isotope ratios in vitamin K1 (phylloquinone) from human plasma by gas chromatography/mass spectrometry. AB - Vitamin K(1) is a fat-soluble vitamin required for the gamma-carboxylation of vitamin K-dependent proteins. Recent work has suggested an important role for vitamin K(1) in bone health beyond its more established function in the control and regulation of blood coagulation. However, current UK recommended intakes do not reflect this recent evidence. The use of stable isotopes provides a powerful tool to investigate vitamin K kinetics, turnover and absorption in man, although published methods have reported difficulties in the extraction and analysis of isotope ratios of vitamin K in human plasma. In this paper, we report a new methodology for the extraction and measurement of isotope ratios in vitamin K(1). Sample clean-up is achieved with liquid-liquid extraction, enzyme hydrolysis with lipase and cholesterol esterase, and solid-phase extraction. Isotopic analysis of the pentafluoropropionyl derivative of vitamin K(1) is performed by gas chromatography/mass spectrometry (GC/MS). The limit of quantitation is equivalent to at least 0.3 nmol/L and the method is demonstrated to be linear over a range of enrichments. This method provides a robust alternative to previous work requiring the use of semi-preparative high-performance liquid chromatography (HPLC). PMID- 16715476 TI - Development of a sensitive and rapid liquid chromatography/tandem mass spectrometry method for the determination of apomorphine in canine plasma. AB - A sensitive, rapid and specific quantitative liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed and validated for the determination of apomorphine (APO) in canine plasma. The analytes were prepared using one-step liquid-liquid extraction, and analyzed on a Waters Symmetry C(18) column interfaced with triple quadrupole tandem mass spectrometer. A mixture of methanol/0.1% formic acid in water (70: 30, v/v) was employed as the isocratic mobile phase. Positive electrospray ionization was utilized as the ionization source. The analyte and clenbuterol (internal standard) were both detected using multiple reaction monitoring (MRM) mode. The limit of detection (LOD) obtained was 0.03 ng/mL. The assay was linear over the concentration range of 0.1-100 ng/mL, and provided good precision (RSD) and good accuracy (RE). The analyte was stable by using antioxidants throughout the whole study. The experimental results show that LC/MS/MS is a rapid and sensitive method to analyze APO in plasma. Finally, the proposed method was successfully applied to a pharmacokinetic study of APO after intranasal administration of 0.5 mg apomorphine to 10 healthy beagle dogs. PMID- 16715477 TI - Hybrid density functional theory study of fragment ions generated during mass spectrometry of 1,3-dioxane derivatives. AB - It was recently reported that the cis,cis and trans,trans diastereoisomers of four 2(r)-R-2,4(R),6(S)-trimethyl-1,3-dioxane derivatives show distinct electron ionization mass spectra. As a possible explanation for this finding, the authors suggested that the ions generated during the mass spectrometry of these compounds could follow different fragmentation patterns that initiate from different ion conformations. In this report, hybrid density functional theory methods have been used to investigate the conformational preference of three ions involved in the mass spectrometry of some 1,3-dioxane derivatives. We found that there is indeed more than one stable ion conformation for each of the investigated ions. Energy profiles along the torsional coordinates connecting the conformers are presented, and factors influencing the relative stability of ion conformations are discussed. PMID- 16715475 TI - Identification of the aromatase inhibitors anastrozole and exemestane in human urine using liquid chromatography/tandem mass spectrometry. AB - Anastrozole (2,2'-[5-(1H-1,2,4-triazol-1-ylmethyl)-1.3-phenylene]bis(2 methylpropionitrile)) and exemestane (6-methylenandrostan-1,4-diene-3,17-dione) are therapeutically used to treat hormone-sensitive breast cancer in postmenopausal women. For doping purposes they may be used to counteract adverse effects of an extensive abuse of anabolic androgenic steroids (gynaecomastia) and to increase plasma testosterone concentrations. Excretion study urine samples and spot urine samples from women suffering from metastatic breast cancer, being treated with anastrozole or exemestane, were collected and analyzed to develop/optimize a detection system for anastrozole and exemestane to allow the identification of athletes who do not comply with the internationally prohibited use of these cancer drugs. The assay was based on liquid-liquid extraction after enzymatic hydrolysis following liquid chromatography/tandem mass spectrometry (LC/MS/MS). Anastrozole, exemestane and its main metabolite (17 dihydroexemestane) were identified in urine by comparison of mass spectra and retention times with respective reference substances. An assay validation for the analysis of anastrozole and exemestane was performed regarding lower limits of detection (anastrozole: 0.02 ng/mL; exemestane: 3.1 ng/mL; dihydroexemestane: 0.5 ng/mL), interday precisions (6.6-11.1%, 4.9-9.1% and 5.6-8.3% for low [10 ng/mL], medium [50 ng/mL] and high [100 ng/mL] concentration) and recoveries (ranged from 85-97%). PMID- 16715478 TI - High-throughput quantification of perindopril in human plasma by liquid chromatography/tandem mass spectrometry: application to a bioequivalence study. AB - A simple, sensitive and rapid high-performance liquid chromatography/positive ion electrospray tandem mass spectrometry method was developed and validated for the quantification of perindopril in human plasma. Following liquid-liquid extraction, the analytes were separated using an isocratic mobile phase on a reversed-phase column and analyzed by mass spectrometry in the multiple reaction monitoring mode using the respective [M+H](+) ions, m/z 369/172 for perindopril and m/z 417/234 for the internal standard. The method exhibited a linear dynamic range of 0.1-100 ng/mL for perindopril in human plasma. The lower limit of quantification was 0.1 ng/mL with a relative standard deviation of less than 6.1%. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. A run time of 2.0 min for each sample made it possible to analyze more than 450 human plasma samples per day. The validated method has been successfully used to analyze human plasma samples for application in pharmacokinetic, bioavailability and bioequivalence studies. PMID- 16715479 TI - Simultaneous determination of low levels of methotrexate and cyclophosphamide in human urine by micro liquid chromatography/electrospray ionization tandem mass spectrometry. AB - The aim of this study was to develop and validate a novel solid-phase extraction (SPE) liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the simultaneous determination of two antineoplastic drugs, cyclophosphamide (CP) and methotrexate (MTX), in human urine using trophosphamide as internal standard. The method showed good precision and accuracy (mean RSD 2.8% and 0.9%; bias 2.7% and 2.4% for MTX and CP, respectively). The lower limits of detection obtained, 0.2 microg/L(urine) for MTX and 0.04 microg/L(urine) for CP, were lower than the best previously reported values. The use of a 96-well SPE plate for matrix purification ensures a high throughput (50 samples/day), allowing the routine biological monitoring of CP and MTX as measures of occupational exposure at very low levels. PMID- 16715480 TI - Electrospray ionization mass spectrometric detection of hexachlorocyclohexane. PMID- 16715481 TI - Investigation of reaction mechanisms by electrospray ionization mass spectrometry: characterization of intermediates in the degradation of phenol by a novel iron/magnetite/hydrogen peroxide heterogeneous oxidation system. AB - Electrospray ionization (ESI) mass spectrometry (MS) and tandem mass spectrometry (MS/MS) were used to monitor the oxidation of phenol by a novel heterogeneous Fenton system based on a Fe(0)/Fe(3)O(4) composite and H(2)O(2). On-line ESI MS(/MS) shows that this heterogeneous system promotes prompt oxidation of phenol to hydroquinone, which is subsequently oxidized to quinone, other cyclic poly hydroxylated intermediates and an acyclic carboxylic acid. A peroxide-type intermediate, probably formed via an electrophilic attack of HOO(.) on the phenol ring, was also intercepted and characterized. ESI-MS(/MS) monitoring of the oxidation of two other model aromatic compounds, benzene and chlorobenzene, indicates the participation of analogous intermediates. These results suggest that oxidation by the heterogeneous system is promoted by highly reactive HO(.) and HOO(.) radicals generated from H(2)O(2) on the surface of the Fe(0)/Fe(3)O(4) composite via a classical Fenton-like mechanism. PMID- 16715482 TI - Electron ionization mass spectrometric study of N-substituted hydrazones of isomeric hydroxybenzaldehydes and isomeric pyridinecarboxaldehydes bearing an N (E)-stilbenyloxyalkylcarbonyltryptophyl substituent. PMID- 16715484 TI - Trans-cultural comparison of disordered eating in Korean women. AB - OBJECTIVE: The presence of eating disorders in non-Western cultures is often attributed to the export of Western ideals. This study examines this hypothesis by comparing disordered eating attitudes and behaviors in Korean women with differing levels of exposure to Western culture. METHOD: Second-generation Korean Americans (n=167) and Korean immigrants (n=37) completed the Eating Attitudes Test (EAT-26), and data from native Koreans (n=937) were obtained from a previous epidemiological study, using a Korean-translated version of the EAT-26 (K-EAT 26). Korean-American and immigrant women completed the Suinn-Lew Asian Self Identity Acculturation Scale (SL-ASIA). RESULTS: Korean-American women scored significantly lower on the EAT-26 than Korean immigrants and native Koreans, who did not differ from each other. Korean-Americans were more Western-oriented than Korean immigrants, and acculturation levels were not correlated with EAT-26 scores in either group. CONCLUSION: This study supports the importance of native cultural factors in the development of eating disorders in non-Western contexts. PMID- 16715483 TI - Evaluation of the in vivo receptor occupancy for the behavioral effects of cannabinoids using a radiolabeled cannabinoid receptor agonist, R [125/131I]AM2233. AB - G-protein coupled receptors exist in both high and low agonist affinity conformations, with tracer levels of agonist radioligands preferentially binding to the former. The goal of the present study was to characterize the in vivo binding of the aminoalkyindole-based, CB1 receptor agonist, R-[125/131I]AM2233 ((2-[125/131I]iodo-phenyl)-[1-(1-methyl-piperidin-2-yl-methyl)-1H-indol-3-yl] methanone), and to use this radiotracer to selectively measure the receptor occupancy by the related CB1 receptor agonist, WIN55212-2, to the agonist preferring affinity state of the receptor. In mouse locomotor assays, both WIN55212-2 and AM2233 (racemic) produced an approximately 60% reduction in activity at 1 mg/kg, (i.v.) and completely inhibited activity at 3 mg/kg, confirming their agonist nature. In ex vivo autoradiography, preferential uptake of R-[131I]AM2233 was apparent in CB1 receptor-rich areas, including globus pallidus, substantia nigra, striatum, cerebellum, and hippocampus. Overall brain uptake of R-[131I]AM2233 was 1.3% injected activity/g at 5 min in mice. Coinjection of 3 mg/kg (i.v.) SR141716A, a CB1 receptor antagonist, with R [125I]AM2233 inhibited the radiotracer binding almost to nonspecific levels in the striatum, globus pallidus, and substantia nigra, although residual binding to a non-CB1 receptor remained in the hippocampus. In contrast to the effect of SR141716A, coinjection of 10 mg/kg (i.v.) WIN55212-2, a high dose that produced an immediate and profound immobility and catalepsy in the mice, reduced CB1 receptor-specific binding of R-[125I]AM2233 in CB1 receptor-rich areas by only 21 43%. These observations suggest that the behavioral effects of CB1 receptor agonists are manifested with a relatively small fraction of the agonist preferring affinity state of the receptor occupied. PMID- 16715485 TI - Eating disorders and celiac disease: a case report. AB - OBJECTIVE: Although chronic physical illness may be associated coincidentally with an eating disorder, some clinicians may overlook the possibility that another medical illness may coexist and contribute concurrently to symptoms such as peculiar eating behaviors, restrictive eating, and/or vomiting accompanied by body dissatisfaction. We present a 31-year-old single woman initially diagnosed with an atypical eating disorder. METHOD: After a gastroenterology consultation prompted by the atypical characteristics of her eating disturbance, the diagnosis of celiac disease was established. RESULTS: Cause-and-effect relationships between anorexia nervosa and celiac disease are unclear, and celiac disease may lead to confusion in the differential diagnosis of anorexia nervosa. CONCLUSION: Particularly in atypical cases, and in cases where nausea and bloating are prominent complaints, workup for celiac sprue may reveal the presence of this condition. In such instances, patients may achieve additional relief through the implementation of gluten-free diets. PMID- 16715486 TI - Eating disorder symptomatology among ballet dancers. AB - OBJECTIVE: The current study sought to compare eating disorder symptomatology among ballet dancers and individuals with restricting anorexia nervosa (RAN), bulimia nervosa (BN), and no eating pathology. METHOD: Twenty-nine female ballet dancers completed assessments and were compared with an archival dataset of 26 women with RAN, 47 women with BN, and 44 women with no eating pathology. Eating disorder diagnoses and behaviors were assessed with a semi-structured clinical interview, the Eating Disorder Inventory (EDI), and a weight history interview. RESULTS: Eighty-three percent of dancers met lifetime criteria for AN (6.9%), BN (10.3%), AN+BN (10.3%), or EDNOS (55.0%). Moreover, dancers looked more similar to eating-disordered individuals than to control individuals on measures of eating pathology. CONCLUSION: Despite previous emphasis on the pathology AN, the current findings suggest that dancers frequently engage in binge eating and purging behaviors. Moreover, it appears that their pathology is as severe as that of non-dancing women with eating disorders. PMID- 16715487 TI - First International Meeting on "Stem Cell Applications in the Craniofacial Region". AB - During the 1st International Meeting on "Stem Cell Applications in the Craniofacial Region" promoted in Naples (Italy), invited researchers presented theirwork and the most innovative methods regarding stem cells (SCs) and their application to the craniofacial region of the human body. In addition, some researchers showed their case-reports on craniofacial reconstruction using either osteo-distraction or reconstruction surgical methods. The aim of this biannual meeting is to stimulate discussion, improve knowledge and promote scientific collaboration among basic and clinical scientists in the main topics of SC use in therapy. A summary of this meeting is given. PMID- 16715488 TI - DSM-IV threshold versus subthreshold bulimia nervosa. AB - OBJECTIVE: The purpose of the present work is to determine whether bulimia nervosa (BN) and eating disorder not otherwise specified, BN type (EDNOS-BN) were qualitatively distinct in terms of eating and general psychopathology. METHOD: This study presents a comparison of 138 women with BN and 57 with EDNOS-BN from a multisite study on eating-related and general psychopathology measures. RESULTS: Although women with BN reported higher lifetime history rates of anorexia nervosa, greater binge eating and vomiting frequency, and more eating concerns, no significant differences were observed between groups on measures of perfectionism, impulsivity, obsessive-compulsiveness, anxiety, depressive symptomatology, or alcohol/substance problems. Based on the partial eta2 values, the distinction between BN and EDNOS-BN accounted for <5% of the criterion variance in general psychopathology measures. Post hoc analyses comparing EDNOS BN with objective bulimic episodes (OBEs; n=34) versus no OBEs (n=23) found greater EDEQ-4 Restraint subscale scores for EDNOS-BN without OBEs. However, there was no significant difference on the EDEQ-4 Eating Concern subscale between the two EDNOS-BN subgroups. CONCLUSION: The findings highlight the clinical significance of BN partial syndrome and prompt reevaluation of existing BN diagnostic boundaries. Post hoc analyses also underscore the need for greater differentiation within EDNOS. PMID- 16715489 TI - Strategic memory in adults with anorexia nervosa: are there similarities to obsessive compulsive spectrum disorders? AB - OBJECTIVE: There is growing interest in the relationship between anorexia nervosa (AN) and obsessive-compulsive (OC) spectrum disorders (e.g., OCD, body dysmorphic disorder [BDD]). Previous neuropsychological investigations of OC spectrum disorders have identified problems with the efficient use of strategy on complex measures of learning and memory. This study evaluated nonverbal strategic memory in AN outpatients using an approach previously applied to OC spectrum disorders. METHOD: Eighteen patients with AN and 19 healthy control participants completed the Rey-Osterrieth Complex Figure Test (RCFT), a widely used measure of nonverbal strategic planning, learning, and memory. RESULTS: Individuals with AN differed significantly from healthy controls in the organizational strategies used to copy the RCFT figure, and they recalled significantly less information on both immediate and delayed testing. Multiple regression analyses indicated that group differences in learning were mediated by copy organizational strategies. CONCLUSION: These results are identical to study findings in OCD and BDD, indicating important shared neuropsychological features among AN and these OC spectrum disorders. As in OCD and BDD, the essential cognitive deficit in AN was impaired use of organizational strategies, which may inform our understanding of the pathophysiology of AN and potentially offer treatment implications. PMID- 16715490 TI - Exocytotic release of creatine in rat brain. AB - The guanidino compound creatine has been shown to occur throughout the brain affecting energy metabolism and mental performance and to act at central GABAA receptors as a partial agonist. Therefore, we examined the possibility that creatine may in fact represent a neuromodulator that is released in the brain in an action-potential dependent manner. To that end, we studied the uptake of [3H]creatine and its electrically evoked release from superfused rat brain slices as well as the evoked release of endogenously synthesized creatine. [3H]creatine was accumulated in neocortex slices in a Na+-dependent manner, consistent with the involvement of the Na+-dependent SLC6A8 creatine transporter. Most importantly, the electrically evoked release of [3H]creatine from neocortex slices (like that from caudate putamen and hippocampus slices) as well as the evoked release of endogenous (unlabeled) creatine was abolished when Ca2+ was omitted from the superfusion medium or in the presence of the Na+-channel blocker tetrodotoxin (TTX). Moreover, blockade of K+-channels by 4-aminopyridine (4-AP) strongly enhanced the electrically evoked release of [3H]creatine as well as that of endogenous creatine. These in vitro data indicate that creatine is not only synthesized and taken up by central neurons, but also released in an action potential dependent (exocytotic) manner, providing strong evidence for its role as a neuromodulator in the brain. PMID- 16715491 TI - Is psychological stress in man associated with increased striatal dopamine levels?: A [11C]raclopride PET study. AB - In rodents, stress causes rapid increases in extracellular dopamine (DA) concentration in cortical and subcortical brain regions, and positron emission tomography (PET) studies in healthy humans have suggested psychological and pharmacological stressors are associated with increased DA concentration in the striatum. In this experiment, we measured the effect of stress, induced by difficult mental arithmetic, on [11C]raclopride binding in order to index striatal DA release. To refine measurements and facilitate interpretation of results a combination of head movement correction, a carefully designed control condition and bolus infusion administration of [11C]raclopride were employed. Fourteen healthy volunteers were scanned using [11C]raclopride PET. Physiological and psychological responses to the task were consistent with a stress response with changes in cardiovascular, hormonal, and subjective state indices. No change of ventral or dorsal striatal [11C]raclopride binding was found in the stress condition compared to nonstress. This negative result suggests that significant DA release does not occur in the striatum in healthy humans after mild, psychological stress. PMID- 16715492 TI - Morphine withdrawal syndrome and its prevention with baclofen: Autoradiographic study of mu-opioid receptors in prepubertal male and female mice. AB - Although the expression of the morphine (MOR) withdrawal syndrome is more marked in male mice than in females, we have demonstrated that the GABAB agonist baclofen (BAC) is able to attenuate MOR withdrawal signs in either sex. In order to extend these previous observations, the aim of the present study was to evaluate the mu-opioid receptor labeling in various brain areas in mice of either sex, during MOR withdrawal and its prevention with BAC. Prepubertal Swiss-Webster mice were rendered dependent by intraperitonial (i.p.) injection of MOR (2 mg/kg) twice daily for 9 days. On the 10th day, dependent animals received naloxone (NAL; 6 mg/kg, i.p.) 60 min after MOR, and another pool of dependent mice received BAC (2 mg/kg, i.p.) previous to NAL. Thirty minutes after NAL, mice were sacrificed and autoradiography with [3H]-[D-Ala2, N-Me-Phe4, -glycol5] enkephalin (DAMGO) was carried out on mice brains at five different anatomical levels. Autoradiographic mapping showed a significant increase of mu-opioid receptor labeling during MOR withdrawal in nucleus accumbens core (NAcC), caudate putamen (CPu), mediodorsal thalamic nucleus (MDTh), basolateral and basomedial amygdala, and ventral tegmental area vs. respective control groups in male mice. In contrast, opiate receptor labeling was not significantly modified in any of the brain areas studied in withdrawn females. BAC reestablished mu-opioid receptor binding sites during MOR withdrawal only in NAcC of males, and a similar tendency was observed in CPu and MDTh, even when it was not statistically significant. The sexual dimorphism observed in the present study confirms previous reports indicating a greater sensitivity of males in response to MOR pharmacological properties. The present results suggest that the effect of BAC in preventing the expression of MOR withdrawal signs could be related with the ability of BAC to reestablish the mu-opioid receptor labeling in certain brain areas. PMID- 16715494 TI - Antipsychotic pathway genes with expression altered in opposite direction by antipsychotics and amphetamine. AB - To develop a new strategy for identifying possible psychotic- or antipsychotic related pathway genes, rats were treated with clinical doses of haloperidol and clozapine for 4 days, and the altered expression of genes was compared with the genes altered in expression after amphetamine sensitization. The objective was to identify genes with expression altered in the same direction by haloperidol and clozapine but in the opposite direction in the amphetamine-sensitized rat striatum. These criteria were met by 21 genes, consisting of 15 genes upregulated by amphetamine, and 6 genes downregulated by amphetamine. Of the 21 genes, 15 are not presently identified, and only 3 genes (cathepsin K, GRK6, and a gene with accession number AI177589) are located in chromosome regions known to be associated with schizophrenia. PMID- 16715493 TI - High resolution spatial mapping of nicotine action using pharmacologic magnetic resonance imaging. AB - Nicotine is one of the most addictive substances known. To better understand the mechanisms of action, we mapped the regional brain response to nicotine administration using pharmacologic magnetic resonance imaging (phMRI) in rats. We measured the regional response of relative cerebral blood volume (rCBV) in rats to a challenge of 0.07 mg/kg (0.43 micromol/kg) of nicotine. The areas of the brain with significant and reproducible changes in the rCBV response were (in descending order of magnitude) infralimbic cortex, hippocampus (subiculum), agranular insular/pyriform cortex, visual cortex, interpeduncular area, nucleus accumbens, cingulate cortex, thalamus, and septum. This pattern of response is consistent with stimulation of both cholinergic and dopaminergic neuronal pathways and is consistent with the known behavioral properties of nicotine. The peak CBV response to nicotine occurred between 9 and 13 min depending upon brain region, and the average full width half-maximum of the rCBV response was 27 min. The high spatial and temporal resolution of the phMRI technique lends itself well to further, more detailed, studies of nicotine dynamics. PMID- 16715495 TI - Reduced basal and ethanol stimulation of striatal extracellular dopamine concentrations in dopamine D2 receptor knockout mice. AB - The present study was undertaken to examine the role of the dopamine (DA) D2 receptor in the ethanol-evoked DA response in the ventral striatum. We performed microdialysis experiments using the D2 null mutant and wild-type controls and measured the effect of an intraperitoneal (i.p.) injection of either saline or ethanol (2 g/kg) on dialysate DA concentrations in the ventral striatum. Dialysate ethanol concentrations were also determined in the samples from the ventral striatum. In addition, the effects of quinpirole, a D2/D3 agonist, were examined in both the ventral and dorsal striatum. Basal dialysate concentrations of DA were significantly reduced in both the ventral and dorsal striatum of the D2 knockouts compared with wild-type controls. Ethanol administration significantly enhanced ventral striatal DA in both groups, but the increase in dialysate DA concentration was 3.5-fold higher in the wild-type controls. The time course of dialysate ethanol concentrations was similar in the two groups. Saline injection did not alter DA concentrations in either the ventral or dorsal striatum. However, quinpirole (0.3 mg/kg) administration significantly depressed striatal dialysate DA concentrations in the wild-type mice, but not in the D2 knockouts. The results suggest that the D2 receptor is necessary for normal development and regulation of striatal extracellular DA concentrations, but the mechanism for this alteration is unclear. In addition, the blunted ethanol-evoked DA response in the D2 knockouts may contribute, in part, to some of the behavioral deficits previously observed in response to ethanol. PMID- 16715496 TI - Inhibition of the glycine transporter GlyT-1 potentiates the effect of risperidone, but not clozapine, on glutamatergic transmission in the rat medial prefrontal cortex. AB - Clinical studies suggest that the efficacy of the atypical antipsychotic drug (APD) risperidone (but not clozapine) can be augmented by adjunctive treatment with agonists at the glycine site of the N-methyl-D-aspartate (NMDA) receptor. By using intracellular recording, we have investigated the effect of the glycine transporter-1 (GlyT-1) inhibitor N [3-(4'-fluorophenyl)-3-(4'phenylphenylphenoxy) propyl] sarcosine (NFPS) on NMDA-induced currents in pyramidal cells of the medial prefrontal cortex (mPFC), both when given alone and in combination with either risperidone or clozapine. Both risperidone and clozapine enhanced the NMDA induced currents. The concentration-response curves were biphasic, and the maximal effect of clozapine on the NMDA-induced currents was significantly larger than the maximal effect of risperidone. NFPS also significantly potentiated the NMDA-induced currents, when given alone. Moreover, NFPS (1 microM) augmented the effect of both the maximal (20 nM), and a submaximal (10 nM), concentration of risperidone. In contrast, NFPS did not potentiate either the effect of the maximal (100 nM) or a submaximal (80 nM) concentration of clozapine on the NMDA induced currents. These data may explain the beneficial clinical results of using glycine reuptake antagonists as adjuvant treatment to risperidone. Our findings also suggest that risperidone and clozapine may affect NMDA receptor-mediated neurotransmission differently in the mPFC. PMID- 16715497 TI - Homologous down-regulation of histamine H3 receptors in rat striatal slices. AB - Preincubation of striatal slices with the selective histamine H3-receptor agonist immepip (100 nM) decreased the specific binding of N-alpha-[methyl-3H]-histamine ([3H]-NMHA) to membranes obtained from the treated slices. The binding decrease was significant after 5 min, remained at similar reduced levels between 5- and 30 min incubations with agonist, and only a partial recovery was observed after 90 min washout (34, 41, and 44% at 90, 120, and 150 min, respectively). Saturation analysis showed a significant decrease in both receptor density (-44% +/- 9%) and affinity (dissociation constant, Kd 1.15 +/- 0.23 nM from 0.59 +/- 0.17 nM). The effect of immepip was mimicked by histamine and the H3 agonists imetit and R alpha-methylhistamine, and was blocked by the H3 antagonist thioperamide. The reduction in [3H]-NMHA binding was fully and partially prevented by incubation at 4 degrees C and in hypertonic medium, respectively, but not by the endocytosis inhibitor phenylarsine oxide (10 microM). None of the following protein kinase inhibitors, Ro-318220 and Go-6976 (PKC), H-89 (PKA) and staurosporine (general inhibitor) prevented the effect of immepip. In [3H]-adenine-labeled slices the preincubation with immepip (100 nM, 15 min) prevented the inhibitory effect of H3 receptor activation on forskolin-induced [3H]-cAMP accumulation (99% +/- 9% vs. 76% +/- 4% of control values). Taken together our results indicate that agonist binding promotes the down-regulation of striatal H3 receptors resulting in a significant loss of function. PMID- 16715498 TI - [123I]TPCNE--a novel SPET tracer for the sigma-1 receptor: first human studies and in vivo haloperidol challenge. AB - [123I]TPCNE (1(trans-[123I]iodopropen-2-yl)-4-[(4-cyanophenoxy)methyl] piperidine; Ki = 0.67 nM; log P = 3.36) is a novel sigma-1 receptor SPET ligand. In this study, we developed an optimized labeling method for [123I]TPCNE and investigated the kinetics, binding characteristics, and whole-body distribution of this tracer for the first time in humans. We also performed a challenge with the sigma-1 receptor antagonist haloperidol against [123I]TPCNE. Seven healthy volunteers were recruited. Dynamic brain SPET scans were performed following i.v. administration of 185 MBq [123I]TPCNE in all seven subjects. Three of the subjects were given oral haloperidol (2.5 mg) approximately 1 h before the scan. The dynamic data were analyzed with both reversible and irreversible compartmental models.[123I]TPCNE showed high uptake in brain and liver. All non haloperidol-treated subjects showed a high whole-brain uptake (average: 8.7% of injected activity). No significant clearance of the tracer was seen up to 30 h post injection. In the haloperidol-treated subjects, the time-activity curves clearly demonstrated clearance of the tracer from the brain. Regional radioactivity concentrations were reduced by haloperidol from 42% in the cerebellum to 73% in the thalamus.[(123)I]TPCNE demonstrated high brain uptake, with highest binding found in the posterior cingulate. A region in which binding was unaffected by haloperidol pretreatment could not be identified, and the time activity data were best described by an irreversible model. PMID- 16715499 TI - In vivo characterization of the pharmacokinetics and pharmacological properties of [11C]-(+)-PHNO in rats using an intracerebral beta-sensitive system. AB - This study reports on the binding kinetics and pharmacological characterization of [11C]-(+)-PHNO ((+)-4-propyl-3,4,4a,5,6,10b-hexahydro-2H-naphtho[1,2 b][1,4]oxazin-9-ol), a promising agonist radiotracer for in vivo evaluation of the D2-receptor. Its in vivo kinetics were monitored in rat striatum and cerebellum using a beta-sensitive Microprobe system. Control studies showed that [11C]-(+)-PHNO binding was reversible and reached a peak time equilibrium of specific binding in striatum 30 min after radiotracer injection. The binding potential (BP) calculated by the simplified reference tissue model was 3-fold higher than that measured with [11C]-(-)-NPA (2.14 +/- 0.50 vs. 0.66 +/- 0.01, respectively). In contrast, the methyl analog of (+)-PHNO, [11C]-(+)-MHNO, which displayed promising D2-agonist properties in vitro, showed no specific binding in the striatum in vivo. [11C]-(+)-PHNO binding was totally blocked by raclopride (1 mg/kg; i.v.) and 97% displaced by NPA (2 mg/kg; i.v.) suggesting that [11C]-(+) PHNO was specific for the high affinity states of D2/D3-receptors. However, (+) PHNO (1 mg/kg; i.v.) totally blocked and displaced [11C]-raclopride binding in striatum. Thus, (+)-PHNO at high concentrations might be able to bind to the low affinity states of D2/D3-receptors. After an amphetamine pretreatment (2 mg/kg; i.v.), a 69% decrease in BP value (P < 0.05) was observed for [11C]-(+)-PHNO indicating that its binding was highly sensitive to variations of endogenous DA. These results substantiate the use of [11C]-(+)-PHNO as an agonist radiotracer for D2-imaging. The sensitivity of its binding to competition with endogenous DA suggests an association with the subset of high affinity state D2-receptors. PMID- 16715500 TI - Early activation, motility, and homing of neonatal microglia to injured neurons does not require protein synthesis. AB - Neuronal injury in CNS tissues induces a rapid activation and mobilization of resident microglia (MG). It is widely assumed that changes in gene expression drive the morphological transformation of MG and regulate their mobilization during activation. Here, we used acutely excised neonatal rat brain slices to test whether the morphological transformation and homing of MG to injured neurons requires gene expression and de novo protein synthesis. Traumatic injury during excision of live brain tissue slices induces a rapid and transient translocation of a transcription factor, NF-kappaB, to nuclei in MG. This is followed within 4 8 h by an increase in immunolabeling for cell adhesion molecules and lysosomal proteins, accompanied by changes in cell morphology. Application of anisomycin, a protein synthesis inhibitor, prevents the increase in immunolabeling for markers of MG activation but not the morphological transformation. Confocal time-lapse imaging in live tissue slices indicates that MG cell motility (branch extension and retraction) and locomotion are unaffected by anisomycin at early postinjury time-points (<4 h), while at later time-points (4-8 h postinjury) MG locomotion but not motility is inhibited. Thus, activated MG rapidly localize to injured pyramidal neuron cell bodies by 4-h postinjury, even in the presence of anisomycin. Moreover, this early MG activation and homing to injured neurons is unaffected in tissue slices from beta2 integrin deficient mice. These results indicate that gene activation and new protein synthesis coincide with, but are not necessary for, the rapid morphological transformation and early migration dependent homing of activated MG to injured neurons in CNS tissues. PMID- 16715501 TI - Treatment of infective endocarditis with recombinant tissue plasminogen activator. AB - Infective endocarditis (IE) caused by microbial infection is virtually always fatal if untreated. High-dose and long-term antibiotic treatment is required to eradicate microorganisms. If increased risk of embolic events, persistent infection, and progressive cardiac failure are present, surgery is indicated. However, surgery can carry an increased risk of mortality and morbidity in critically ill children of whom other treatment options such as administering, a thrombolytic agent; recombinant tissue plasminogen activator (r-tPA) could be an alternative choice. Here, we report a 14-year-old male with Down syndrome and acute myeloblastic leukemia, diagnosed with IE characterized by two large vegetations on aortic and mitral valves, who was successfully treated with r-tPA. PMID- 16715502 TI - A new serine/threonine protein kinase, Omphk1, essential to ventral body wall formation. AB - Here, we report a new serine/threonine protein kinase of the SNF1 subfamily Omphk1. Two Omphk homologues exist in each vertebrate species, and one homologue exists in Drosophila and Caenorhabditis elegans; the kinase domain is highly conserved among these homologues, and several domains are conserved among vertebrate Omphk. Omphk1 expression dynamically changes in the developing central nervous system, is found ubiquitously in epidermis, and is present uniquely in several other tissues. Its expression is also found in each tissue associated with the ventral body wall closure: the primary body wall composed of primitive ectoderm and each component of the secondary body wall. Concomitantly, its null mutant exhibits omphalocele with a failure in closure of the secondary body wall. There are no apparent gross morphological defects in brain, however, despite the unique Omphk1 expression in this tissue. PMID- 16715503 TI - Accessory chemosignaling mechanisms in primates. AB - Accessory olfaction is defined as the chemoreceptive system that employs the vomeronasal complex (VNC) and its distinct central projections to the accessory olfactory bulb (AOB) and limbic/cortical systems. Comparisons of the structural and functional features of primate accessory olfaction can now be made at many levels. Advances in the understanding of molecular mechanisms of odorant transfer and detection, physiological analyses of signal processing, and appreciation of ontogenetic timetables have clarified the contribution of accessory chemoreception to the sensory map. Two principal functions dominate: the decoding of social information through the uptake of signals (often fluid-borne), and the provision of an essential pathway for the "migration" of presumptive neurocrine (GnRH) cells from the olfactory placode to the hypothalamus. VN "smelling" (vomerolfaction) is now seen to overlap with primary olfaction. Both systems detect signal compounds along the spectrum of volatility/molecular weight, and neither is an exclusive sensor. Both main and accessory chemoreception seem to require collaborative molecular devices to assist in odorant transfer (binding proteins) and (for the VNO) signal recognition (MHC1 proteins). Most adaptive selective features of primate chemocommunication variously resemble those of other terrestrial mammals. VN function, along with its genome, has been maintained within the Strepsirrhines and tarsiers, reduced in Platyrrhines, and nearly extinguished at the Catarrhine up to hominin levels. It persists as an intriguing ancient sense that retains key features of past evolutionary events. PMID- 16715504 TI - Genetic basis of olfactory communication in primates. AB - The genes involved in olfactory communication in mammals via the vomeronasal system are summarized, and studies investigating these genes in primates are reviewed. Only five potentially functional vomeronasal receptor genes (V1RL s) have been found in humans, and only one of these (V1RL1) has been studied in other primates. V1RL1 has become a pseudogene repeatedly during primate evolution, but patterns of natural selection on primate V1RL genes demonstrate that this gene family diverged under natural selection throughout at least part of primate evolution. Evolution of the TRP2 gene, which encodes for an ion channel that is important in vomeronasal organ (VNO) signalling, strongly suggests that this signalling function was lost in ancestral Catarrhines. Overall, much work remains to be done to elucidate the repertoire of genes that are involved in pheromonal communication, particularly in Strepsirhines. Such studies promise unique insights into the evolution of this modality. PMID- 16715505 TI - The neglected sense-olfaction in primate behavior, ecology, and evolution. AB - This special issue emerged from a symposium held during the 20th Congress of the International Primatological Society in Torino, Italy, in August 2004. The symposium brought together scientists studying several different aspects of olfaction in primates. The topics addressed ranged from the morphology and physiology of the sensory apparatus, the genetics and chemistry of olfactory signals and the use of such signals in primate communication, to a comparative analysis of the role of olfaction in neural evolution. The papers in this issue reflect a surge of interest in diverse aspects of olfaction-an interest that has been stimulated by the more rigorous theoretical approaches and new techniques that have recently become available. This introduction briefly reviews past research on primate olfaction, summarizes the scope of this special issue, and provides a somewhat speculative glimpse of the future. PMID- 16715506 TI - Olfactory evolution and behavioral ecology in primates. AB - Primates are usually thought of as "visual" mammals, and several comparative studies have emphasized the role of vision in primate neural and sociocognitive specialization. Here I explore the role of olfactory systems, using phylogenetic analysis of comparative volumetric data. The relative sizes of the main olfactory bulb (MOB) and accessory olfactory bulb (AOB) tend to show different evolutionary patterns in accordance with their different functions. Although there is some evidence of correlated evolution of the two systems, this is apparent in only one clade (the strepsirhines). As predicted, the MOBs correlate predominantly with ecological factors (activity period and diet), while the AOBs correlate with social and mating systems. Related olfactory structures (i.e., the piriform cortex and amygdala) exhibit correlated evolution with the AOBs but not with the MOBs, and the corticobasolateral part of the amygdala exhibits a correlation with social group size in platyrrhines similar to that observed for the AOB. These social system correlations support the idea that there is an olfactory dimension to the concept of the social brain. PMID- 16715507 TI - Olfactory signals and the MHC: a review and a case study in Lemur catta. AB - The major histocompatibility complex (MHC) is the most polymorphic genetic system known in vertebrates. Decades of research demonstrate that it plays a critical role in immune response and disease resistance. It has also been suggested that MHC genes influence social behavior and reproductive phenomena. Studies in laboratory mice and rats report that kin recognition and mate choice are influenced by olfactory cues determined at least in part by an individual's MHC genes. This issue has stimulated intense but controversial research. However, work in this field has only been carried out in rodents and humans. Thus far, no study has directly investigated the relationship between olfactory cues and MHC genotype in nonhuman primates. Furthermore, other genetic loci, including those linked to the MHC, have not been ruled out as the primary influence on odor profiles. To explore the relationship between individual odor profiles and MHC alleles, we are studying ring-tailed lemurs (Lemur catta). These animals are an ideal model species because they are extremely scent-oriented and their behaviors suggest that olfactory signals form an important part of their intra- and intergroup communication systems. Individual odor profiles from tail and scent gland samples were generated for six males using gas chromatography mass spectrometry (GC-MS). MHC genotypes were identified using polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE). The GC-MS analyses demonstrated a difference between profiles obtained from tail and scent gland samples. Although our sample size is relatively small and statistical significance could not be obtained, our analyses suggest a relationship between MHC and concentrations of volatile compounds. While these results are preliminary, they support the need for further studies of the MHC and olfactory signals in lemurs and other primates. PMID- 16715508 TI - Individual olfactory signatures in common marmosets (Callithrix jacchus). AB - Individual discrimination is likely a prerequisite for most primate social interactions. Olfactory cues are one set of stimuli used by primates to discriminate between individuals. Despite the importance of these olfactory signatures, there is little published research assessing the existence or function of individually unique odors among primates. This review systematically assesses behavioral and biochemical aspects of individual odors in a New World primate, the common marmoset (Callithrix jacchus). There are three objectives to this review: First, behavioral evidence for odors unique to the individual is evaluated in the context of results demonstrating that marmosets are able to discriminate between the scents from a familiar and a novel individual conspecific in behavioral bioassays under a variety of conditions. Second, biochemical evidence for individual scent signatures is debated with reference to studies examining qualitative and quantitative differences between the chemical compositions of scent-mark pools from adult females. A combined gas chromatography (GC) and mass spectrometry (MS) analysis demonstrated that each female had a unique ratio of highly volatile chemicals in the scent mark that could affect individual discrimination. Finally, the possible adaptive significance of individual odors in marmosets is debated. Individual odors may play a key role in regulating both female intrasexual competition and intersexual communication by providing a basis for the assessment of individual quality. PMID- 16715509 TI - Scent marking as resource defense by female Lemur catta. AB - Because ring-tailed lemurs (Lemur catta) are a female-dominant, female philopatric species in which the females provide the majority of parental care and troop defense, resource defense is a possible function of female lemur scent marking. To test this hypothesis, I conducted three studies. First, I presented captive, individually housed females with a series of samples of female scent, each from a different female, to determine whether they would respond to those samples and discriminate between them. Second, I reanalyzed data from a focal animal study of four females in two adjacent troops in Berenty Reserve, Madagascar, to determine female marking rates before, during, and after the mating season, and to clarify the relationship among positions of feeding, intertroop defense, and scent marking. The third study was based on ad libitum observations of the sniffing and marking behavior of a troop in Berenty Reserve during a year when they traveled far out of their home range. The females in study 1 investigated female scent samples but provided no evidence that they discriminated between them. In study 2 the wild females marked throughout the study and did not limit their marking to the mating season. They deposited significantly more of their marks in a zone of confrontation with adjacent troops, where they also did the majority of their feeding, and they increased their rate of marking during agonistic intertroop confrontations. The females determined the positions of their scent marks and deposited the first mark in the majority of countermarking sequences. When the females traveled out of their defended range in study 3, they significantly decreased their rate of marking and increased their rate of sniffing spots but not marking them. All evidence gathered so far supports the hypothesis that one function of female ring-tailed lemur scent marking is to provide intergroup information that is then used to reinforce the border of the defended resource. PMID- 16715511 TI - Scent marking strategies of New World primates. AB - This paper reviews recent findings on scent-marking behavior in wild New World primates. These findings are used to examine three functional hypotheses: territoriality, regulation of social and reproductive dominance, and mating competition/mate attraction. Available data provide little evidence for a territorial function of scent-marking behavior in New World primates. Evidence for a function in the regulation of social and reproductive dominance is ambiguous. The patterns of scent marking found so far (i.e., rates of scent marking according to sex and social or reproductive status, and the spatial distribution of scent marks), and the sparse information on responses to scent marks are consistent with a hypothesized function in intrasexual competition and intersexual mate choice. Suggestions for future research are made. PMID- 16715510 TI - Scent marking in sifaka: no one function explains it all. AB - Various hypotheses to explain possible functions of scent-marking have been put forth and basically fall into five categories: territorial demarcation, ownership of resources, mate attraction, noncombative fighting, and self-advertisement. Verreaux's sifaka (Propithecus verreauxi verreauxi), like many prosimians, use scent-marking as one form of communication. The goal of this study was to determine the function of scent-marking in sifaka. All-occurrence of scent-marks, scent-mark context, and scent-mark style were collected on 23 sifaka in the Kirindy Forest of western Madagascar for 7 months (September 2001-March 2002). Scent-mark rates were collected using continuous focal animal sampling from November 2000-March 2002. Home range data were collected using monthly censuses and instantaneous focal sampling throughout the 17 months. Scent-marking behavior was exhibited almost exclusively by adults. Scent-mark rates were not related to the number of resident adult males, number of resident adult females, rank, or group size. The majority of scent-marks occurred in the perimeter of the home range; however, less than a quarter of the home range was used by only a single group. Sifaka did not preferentially mark food trees, mark during the mating season, or mark during intergroup encounters. The results from this study indicate that none of the five hypothesized functions best explains all scent marking in Verreaux's sifaka. Rather, scent-marking may serve different functions for different individuals. PMID- 16715512 TI - Scent marking in two western Amazonian populations of woolly monkeys (Lagothrix lagotricha). AB - We describe patterns of scent marking observed in two wild populations of lowland woolly monkeys that were the subjects of long-term studies in the westernmost portion of the Amazon basin. The woolly monkeys engaged primarily in two types of scent marking: chest rubbing and anogenital rubbing. In both study populations, males and females performed both types of scent marking, but males chest-rubbed more commonly than females, while females engaged in more anogenital rubbing. We evaluated two nonexclusive hypotheses for the function of scent marking by woolly monkeys: 1) that scent marking is used in sociosexual contexts, and 2) that scent marking is used to convey information about occupancy of or willingness to defend an area from conspecifics in other social groups. We found no association between the occurrence of scent-marking behavior and location within the home range, but did find that scent marking occurred more commonly than expected on days when copulations, mating solicitations, and intergroup encounters were observed. Additionally, mating activity and chest rubbing were highly correlated across the yearly cycle, even when the potentially confounding variable of ripe fruit availability was controlled for. In woolly monkeys, overt male-male competition is rare and female choice is an important part of the mating system. Our results are most consistent with the idea that scent marking plays a role in advertising male quality or competitive ability, and perhaps in coordinating mating activity. PMID- 16715513 TI - Roles of jumonji and jumonji family genes in chromatin regulation and development. AB - The jumonji (jmj) gene was identified by a mouse gene trap approach and has essential roles in the development of multiple tissues. The Jmj protein has a DNA binding domain, ARID, and two conserved jmj domains (jmjN and jmjC). In many diverse species including bacteria, fungi, plants, and animals, there are many jumonji family proteins that have only the jmjC domain or both jmj domains. Recently, Jmj protein was found to be a transcriptional repressor. Several proteins in the jumonji family are involved in transcriptional repression and/or chromatin regulation. Most recently, one of the human members has been shown to be a histone demethylase, and the jmjC domain is essential for the demethylase activity. Meanwhile, more and more evidence indicating that the jumonji family proteins play important roles during development is accumulating. Many proteins in the jumonji family may regulate chromatin and gene expression, and control development through various signaling pathways. Here, we highlight the roles of jmj and jumonji family proteins in chromatin regulation and development. PMID- 16715514 TI - Direct HPLC enantioseparation of chiral aptazepine derivatives on coated and immobilized polysaccharide-based chiral stationary phases. AB - The direct HPLC enantioseparation of Mianserin and a series of aptazepine derivatives is accomplished on polysaccharide-based chiral stationary phases (CSPs). The resolutions are performed on the coated-type Chiralcel OD and Chiralpak AD CSPs and on the first commercially available immobilized-type Chiralpak IA CSP, in normal-phase and polar-organic modes. The complete separation of enantiomers of all racemates investigated was successfully achieved under at least one of CSP/eluent combinations employed. Pure alcohols such ethanol or 2-propanol, with a fixed percentage of DEA added, serve as valuable alternatives to the more common n-hexane-based normal-phase eluents in resolution of Mianserin on the AD CSP. In order to study the chiroptical properties of aptazepine derivatives, chromatographic resolutions are carried out at semipreparative scale using Chiralpak AD and Chiralpak IA as CSPs. Nonconventional dichloromethane-based eluents have permitted to expand the chiral resolving ability of the immobilized Chiralpak IA CSP and to perform mg-scale enantioseparations with an analytical-size column. Assignment of the absolute configuration of the separated enantiomers is empirically established by comparing their chiroptical data with those of structurally related Mianserin. PMID- 16715516 TI - Elucidation of the absolute configuration of JNJ-27553292, a CCR2 receptor antagonist, by vibrational circular dichroism analysis of two precursors. AB - The absolute configurations of two precursors, that is, 1-(3',4'-dichlorophenyl) propanol and 1-(3',4'-dichlorophenyl)-propanamine, of a potent 2-mercapto imidazole CCR-2 receptor antagonist, JNJ-27553292, were determined using vibrational circular dichroism. As a consequence, the absolute configuration of the antagonist itself was also determined. The two precursor compounds were subjected to a thorough conformational analysis and rotational strengths were calculated at the B3LYP/cc-pVTZ level of theory. Based on these data, vibrational circular dichroism spectra were simulated, which in turn were compared with experimental spectra. Agreement between the spectra allowed the assignment of the absolute configuration, which is in agreement with the proposed configuration based on stereospecific reactions on similar compounds. PMID- 16715515 TI - Control of optically active structure of thioether-phthalocyanine aggregates by chiral Pd(II)-BINAP complexes in toluene and at the toluene/water interface. AB - The Pd(II)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (BINAP)-mediated chiral assembly of thioether-derivatised phthalocyanatomagnesium(II) compounds (MgPc(SR)8, SR is benzylthio (SBz) or benzhydrylthio (Bh)) was formed in toluene and at the toluene/water interface, and investigated by means of UV-Vis absorption and circular dichroism (CD) spectroscopy combined with the centrifugal liquid-membrane (CLM) devise. Interfacial tension measurements indicated that, in the presence of PdSO4 in the aqueous phase, BINAP ligand adsorbed as a monolayer forming Pd(II)BINAP2+ complex at the toluene/water interface. UV-Vis absorption spectrum of MgPc(SR)8 in the Q-band region was blue-shifted in toluene upon addition of [Pd(II)BINAP]Cl2, but red-shifted at the toluene/water interface when Pd(II)BINAP2+ was formed at the interface. These results suggested that MgPc(SR)8 Pd(II)BINAP complex formed H-aggregate (face-to-face type) in toluene solution and J-aggregate (end-to-end type) at the toluene/water interface, respectively. Moreover, the specific bisignate CD spectral pattern in both systems indicated that the aggregates of MgPc(SR)8-Pd(II)BINAP complexes were chirally twisted, well controlled by the chirality of BINAP ligand. Very interestingly, the morphology of MgPc(SBz)8-Pd(II)BINAP and MgPc(SBh)8-Pd(II)BINAP aggregates formed at the toluene/water interface were significantly different as the rodlike and the ribbonlike crystalline structure, respectively, as observed by a scanning electron microscopy (SEM). On the basis of these experimental results, we proposed schematic molecular models of the chiral aggregates of MgPc(SR)8 Pd(II)BINAP complexes and demonstrated the specific role of the toluene/water interface. PMID- 16715517 TI - Elucidation of the enantioselective recognition mechanism of a penicillin G acylase-based chiral stationary phase towards a series of 2-aryloxy-2-arylacetic acids. AB - A series of structurally related 2-aryloxy-2-arylacetic acids (1-3, 5-16) together with a thioisostere derivative (4) have been synthesized and characterized by GC-MS and 1H NMR. The designed compounds were analyzed on a Penicillin G Acylase chiral stationary phase (PGA-CSP) and the influence of the structure variations on retention and enantioselectivity was investigated. The chromatographic study includes the direct separation of the enantiomers of the synthesized compounds and the determination of the elution order of selected racemic mixtures. 10 out of 16 racemates were separated; high chromatographic enantioseparation factors (alpha > 2) were achieved for some compounds. For the enantiomers of four compounds whose absolute configuration was known (1, 3, 12, 16), the elution order was R:S with the exception of 2-(4-chloro phenoxy)phenylacetic acid (1), for which the elution order was reversed. Preliminary molecular modeling studies suggest that both polar and charge transfer interactions as well as steric effects play an important role in determining the retention factors and the enantioselectivities observed. PMID- 16715518 TI - Prevalence of BRCA1 genomic rearrangements in a large cohort of Italian breast and breast/ovarian cancer families without detectable BRCA1 and BRCA2 point mutations. AB - The presence of genomic rearrangements of the BRCA1 gene in breast and/or ovarian cancer families has been intensively investigated in patients from various countries over the last years. A number of different rearrangements have been reported by several studies that clearly document the involvement of this mutation type in genetic predisposition to breast and ovarian cancer. Population specific studies are now needed to evaluate the prevalence of genomic rearrangements before deciding whether to include ad hoc screening procedures into standard diagnostic mutation detection approaches. Indeed, the vast majority of the studies have been performed on small, highly selected, sample sets because of the limitations imposed by the laborious technical approaches. Moreover, prevalence figures are likely to differ across different countries according to the ethnic origin of each specific population. Here we analyze a large cohort of 653 Italian probands, negative for BRCA1 and BRCA2 point mutations, gathered from four National Institutions. We report the identification of BRCA1 genomic rearrangements in 12 independent families. Noteworthy, half of the probands carry mutations that recur in more than one Italian family. Considering the whole spectrum of Italian BRCA1 gene rearrangements identified thus far in consecutive patients, we estimate that alterations of this type account for 19% (95% CI: 0.11 < 0.19 < 0.28) of the BRCA1 mutation positive families. We conclude that the search for major genomic rearrangements is essential for an accurate and comprehensive BRCA1 mutation detection strategy in Italy. PMID- 16715519 TI - Complex dependence of substrate stiffness and serum concentration on cell-force generation. AB - Collagen is a widely used biomaterial in tissue engineering. Mechanical stimulation of cell-seeded collagen constructs and its effects on cell orientation, intracellular signaling, and molecular responses have been reported. Our aim was to study the transfer of applied mechanical load to resident cells in 3D collagen constructs. Stainless steel markers were embedded in constructs as reporters of micromovement and uniaxial (0-15%) strain was applied. Cell-seeded collagen constructs were also subjected to (0-15%) uniaxial strain and material responses recorded. The viscoelastic properties of collagen resulted in comparatively small movement of the marker bars relative to gel deformation. Cell seeding density of 1 million/mL had no significant effect on the viscoelastic properties of collagen for the range of strain tested. Our findings indicate that viscoelastic properties of collagen result in minimal force transfer of applied loads as recorded by movement of stainless steel markers. At higher strain rates as collagen got stiffer the movement decreased. These findings indicate that as cell-seeded collagen constructs mature in a bioreactor and become stiffer due to ECM production/deposition, mechanical stimulation will have to be tailored over time to account for increased stiffness of constructs in vitro to elicit predictable and consistent cellular responses. PMID- 16715520 TI - Real-time measurement of cytosolic free calcium concentration in Jurkat cells during ELF magnetic field exposure and evaluation of the role of cell cycle. AB - Extremely low frequency magnetic fields (ELF MF) have been reported to alter a number of cell signaling pathways, including those involved in proliferation, differentiation and apoptosis where cytosolic free calcium ([Ca(2+)](c)) plays an important role. To better understand the biological conditions under which ELF MF exposure might alter [Ca(2+)](c), we measured [Ca(2+)](c) by ratiometric fluorescence spectrophotometry during exposure to ELF MF in Jurkat E6.1 cells synchronized to different phases of the cell cycle. Suspensions of cells were exposed either to a near zero MF (Null) or a 60 Hz, 100 microT sinusoidal MF superimposed upon a collinear 78.1 microT static MF (AC + DC). An initial series of experiments indicated that the maximum increase in [Ca(2+)](c) above baseline after stimulation with anti-CD3 was significantly higher in samples exposed to AC + DC (n = 30) compared to Null (n = 30) with the largest difference in G2-M enriched samples. However, in a second study with G2-M enriched cells, samples treated with AC + DC (n = 17) were not statistically different from Null-treated samples (n = 27). Detailed analysis revealed that the dynamics in [Ca(2+)](c) before and after stimulation with anti-CD3 were dissimilar between Null samples from each study. From the results, we concluded (i) that the ELF MF increased [Ca(2+)](c) during an antibody-induced signaling event, (ii) that the ELF MF effect did not depend to a large degree on cell cycle, and (iii) that a field related change in [Ca(2+)](c) signaling appeared to correlate with features in the [Ca(2+)](c) dynamics. Future work could evaluate [Ca(2+)](c) dynamics in relation to the phase of the cell cycle and inter-study variation, which may reveal factors important for the observation of real-time effects of ELF MF on [Ca(2+)](c). PMID- 16715521 TI - Exposure of murine cells to pulsed electromagnetic fields rapidly activates the mTOR signaling pathway. AB - Murine pre-osteoblasts and fibroblast cell lines were used to determine the effect of pulsed electromagnetic field (PEMF) exposure on the production of autocrine growth factors and the activation of early signal transduction pathways. Exposure of pre-osteoblast cells to PEMF minimally increased the amount of secreted TGF-beta after 1 day, but had no significant effects thereafter. PEMF exposure of pre-osteoblast cells also had no effect on the amount of prostaglandin E(2) in the conditioned medium. Exposure of both pre-osteoblasts and fibroblasts to PEMF rapidly activated the mTOR signaling pathway, as evidenced by increased phosphorylation of mTOR, p70 S6 kinase, and the ribosomal protein S6. Inhibition of PI3-kinase activity with the chemical inhibitor LY294002 blocked PEMF-dependent activation of mTOR in both the pre-osteoblast and fibroblast cell lines. These findings suggest that PEMF exposure might function in a manner analogous to soluble growth factors by activating a unique set of signaling pathways, inclusive of the PI-3 kinase/mTOR pathway. PMID- 16715523 TI - Comment concerning "Do electromagnetic fields interact with electrons in Na,K ATPase?" (Bioelectromagnetics 26:677-683 [2005]). PMID- 16715524 TI - Effects of a 50 Hz magnetic field on Dictyostelium discoideum (Protista). AB - Some studies have demonstrated that a few biological systems are affected by weak, extremely low frequency (ELF) electromagnetic fields (EMFs), lower than 10 mT. However, to date there is scanty evidence of this effect on Protists in the literature. Due to their peculiarity as single-cell eukaryotic organisms, Protists respond directly to environmental stimuli, thus appearing as very suitable experimental systems. Recently, we showed the presence of propionylcholinesterase (PrChE) activity in single-cell amoebae of Dictyostelium discoideum. This enzyme activity was assumed to be involved in cell-cell and cell environment interactions, as its inhibition affects cell aggregation and differentiation. In this work, we have exposed single-cell amoebae of D. discoideum to an ELF-EMF of about 200 microT, 50 Hz, for 3 h or 24 h at 21 degrees C. A delay in the early phase of the differentiation was observed in 3 h exposed cells, and a significant decrease in the fission rate appeared in 24 h exposed cells. The PrChE activity was significantly lower in 3 h exposed cells than in the controls, whereas 24 h exposed cells exhibited an increase in this enzyme activity. However, such effects appeared to be transient, as the fission rate and PrChE activity values returned to the respective control values after a 24 h stay under standard conditions. PMID- 16715525 TI - Phosphorylation and gene expression of p53 are not affected in human cells exposed to 2.1425 GHz band CW or W-CDMA modulated radiation allocated to mobile radio base stations. AB - A large-scale in vitro study focusing on low-level radiofrequency (RF) fields from mobile radio base stations employing the International Mobile Telecommunication 2000 (IMT-2000) cellular system was conducted to test the hypothesis that modulated RF fields induce apoptosis or other cellular stress response that activate p53 or the p53-signaling pathway. First, we evaluated the response of human cells to microwave exposure at a specific absorption rate (SAR) of 80 mW/kg, which corresponds to the limit of the average whole-body SAR for general public exposure defined as a basic restriction by the International Commission on Non-Ionizing Radiation Protection (ICNIRP) guidelines. Second, we investigated whether continuous wave (CW) and wideband code division multiple access (W-CDMA) modulated signal RF fields at 2.1425 GHz induced apoptosis or any signs of stress. Human glioblastoma A172 cells were exposed to W-CDMA radiation at SARs of 80, 250, and 800 mW/kg, and CW radiation at 80 mW/kg for 24 or 48 h. Human IMR-90 fibroblasts from fetal lungs were exposed to both W-CDMA and CW radiation at a SAR of 80 mW/kg for 28 h. Under the RF field exposure conditions described above, no significant differences in the percentage of apoptotic cells were observed between the test groups exposed to RF signals and the sham-exposed negative controls, as evaluated by the Annexin V affinity assay. No significant differences in expression levels of phosphorylated p53 at serine 15 or total p53 were observed between the test groups and the negative controls by the bead-based multiplex assay. Moreover, microarray hybridization and real-time RT-PCR analysis showed no noticeable differences in gene expression of the subsequent downstream targets of p53 signaling involved in apoptosis between the test groups and the negative controls. Our results confirm that exposure to low-level RF signals up to 800 mW/kg does not induce p53-dependent apoptosis, DNA damage, or other stress response in human cells. PMID- 16715526 TI - Integrins may serve as mechanical transducers for low-frequency electric fields. AB - A hypothesis is presented that a transduction mechanism for low frequency electric fields of physiological strength ( approximately 1 V/cm) is the same as that for sinusoidal fluid shear stresses, the force exerted on an integrin. Simple calculations show that the forces exerted on a model integrin by transverse electric fields and fluid shears that produce cellular effects are comparable in magnitude, about 1 fN. The electric force is provided by the interaction of the surface charges on the integrin with the tangential component of the applied field. The mechanical shear force is the transverse fluid drag force exerted on the cylindrical surface of the integrin. Either force is coupled mechanically to the actin cortex within the cell. The mechanical network which exists within a cell and connects a cell to its surroundings would then be directly coupled to an applied electric field. The fundamental transduction mechanism for some electric field effects may then be ultimately mechanical in nature. PMID- 16715527 TI - Modeling normal and altered human erythrocyte shapes by a new parametric equation: application to the calculation of induced transmembrane potentials. AB - We present simple parametric equations in terms of Jacobi elliptic functions that provide a realistic model of abnormal variations in size which maintain the biconcave shape of a normal erythrocyte (anisocytosis) and abnormal variations in shape which maintain the original volume of the erythrocyte (poikilocytosis), as well as continuous deformations from the normal to the altered shapes. We illustrate our results with parameterizations of microcytes, macrocytes, and stomatocytes, and we apply these parameterizations to the numerical calculation of the induced transmembrane voltage in microcytes, macrocytes, and stomatocytes exposed to an external electromagnetic field of 1800 MHz. PMID- 16715528 TI - Oxidative stress effects on the central nervous system of rats after acute exposure to ultra high frequency electromagnetic fields. AB - Mobile telephones and their base stations are an important source of ultra high frequency electromagnetic fields (UHF-EMFs; 800-1800 MHz) and their utilization is increasing all over the world. Epidemiological studies have suggested that low energy UHF-EMFs may have biological effects, such as changes in oxidative metabolism after exposure. Therefore, we have investigated the effect of acute UHF-EMF exposure on non-enzymatic antioxidant defense and lipid and protein oxidative damage in the rat frontal cortex and hippocampus. We have used malondialdehyde (MDA) and carbonyl assays to assess lipid and protein oxidative damages, respectively. No changes in lipid and protein damage, and also in non enzymatic defense were found in frontal cortex or hippocampus. These results suggest that acute UHF-EMF exposure is not able to produce detectable oxidative stress in rats from any age tested. However, more tests using a longer period of exposure and evaluating other tissues are necessary to ensure that there is no health risk associated with the use of mobile phones. PMID- 16715529 TI - 50 Hz magnetic fields of constant or fluctuating intensity: effects on immunocyte hsp70 in the mussel Mytilus galloprovincialis. AB - Effects of a single 30 min exposure to a 50 Hz, 400 microT sinusoidal extremely low frequency magnetic field (ELF MF) on hsp70 expression in immunocytes from the bivalve Mytilus galloprovincialis was investigated using RT-PCR and immunoblot approaches. The results indicate that in M. galloprovincialis immunocytes hsp70 expression was unaltered at both transcriptional and translational level after exposure to constant or fluctuating intensity MFs. PMID- 16715530 TI - Is measurement of nuchal translucency thickness a useful screening tool for heart defects? A study of 16,383 fetuses. AB - OBJECTIVE: To determine the performance of nuchal translucency thickness (NT) measurement as a screening method for congenital heart defects (CHD) among fetuses with normal karyotype. METHODS: An NT measurement was made in 16 383 consecutive euploid fetuses derived from an unselected pregnant population. The cut-offs for increased risk of heart defects, chosen a priori and tested prospectively, were: NT >or= 95th centile for crown-rump length, NT >or= 3 mm, and NT >or= 3.5 mm. The sensitivity and false-positive rate (FPR; 1 minus specificity) of the risk cut-offs and their positive and negative likelihood ratios (+LR and -LR) with regard to CHD were calculated. RESULTS: Among the 16 383 fetuses with an NT measurement there were 127 cases with a diagnosis of heart defect confirmed by cardiac investigations after birth or at autopsy. Of these, 55 defects were defined as major, of which 52 were isolated (no other defects or chromosomal aberrations), corresponding to a prevalence of major heart defects in chromosomally normal fetuses/newborns of 3.3/1000. The sensitivity, FPR, +LR and LR for NT >or= 95th centile with regard to an isolated major heart defect were: 13.5%, 2.6%, 5.2 and 0.9, respectively. For NT >or= 3.0 mm these values were: 9.6%, 0.8%, 12.0 and 0.9, and for NT >or= 3.5 mm they were: 5.8%, 0.3%, 19.3 and 0.9. CONCLUSIONS: NT measurement is a poor screening method for isolated major CHD. A method with a much higher detection rate and with a reasonably low FPR is needed. However, increased NT indicates increased risk of fetal heart defect, and women carrying fetuses with increased NT should be offered fetal echocardiography in the second trimester. PMID- 16715531 TI - Accelerated repair of cortical bone defects using a synthetic extracellular matrix to deliver human demineralized bone matrix. AB - Injectable hydrogel and porous sponge formulations of Carbylan-GSX, a crosslinked synthetic extracellular matrix (ECM), were used to deliver human demineralized bone matrix (DBM) in a rat femoral defect model. A cortical, full-thickness 5-mm defect was created in two femurs of each rat. Six rats were assigned to each of five experimental groups (thus, 12 defects per group). The defects were either untreated or filled with Carbylan-GSX hydrogel or sponges with or without 20% (w/v) DBM. Radiographs were obtained on day 1 and at weeks 2, 4, 6, and 8 postsurgery of each femur. Animals were sacrificed at week 8 postsurgery and each femur was fixed, embedded, sectioned, and processed for Masson's Trichrome staining. The bone defects were measured from radiographs and the fraction of bone healing was calculated. The average fractions of bone healing for each group were statistically different among all groups, and all treatment groups were significantly better than the control group. The Carbylan-GSX sponge with DBM was superior to the sponge without DBM and to the hydrogel with DBM. Histology showed that defects treated with the Carbylan-GSX sponge plus DBM were completely filled with newly generated bone tissue with a thickness comparable to native bone. Carbylan-GSX sponge was an optimal delivery vehicle for human DBM to accelerate bone healing. PMID- 16715532 TI - Tracking chondrocytes and assessing their proliferation with PKH26: effects on secretion of proteoglycan 4 (PRG4). AB - Distinguishing between implanted and host-derived cells, as well as between distinct cell phenotypes, would be useful in assessing the mechanisms of cell based repair of cartilage. The fluorescent tracker dye, PKH26, was previously applied to several cell types to assess proliferation in vitro and to track cells in vivo. The objectives of this study were to assess the utility of PKH26 for tracking chondrocytes from superficial and middle zones and their proliferation, and determine the effects of PKH26 on chondrocyte functions, in particular, proliferation and secretion of Proteoglycan 4 (PRG4). PKH26-labeled and unlabeled superficial and middle zone chondrocytes were plated in either low- or high density monolayer culture and analyzed for retention of PKH26 by flow cytometry and fluorescence microscopy at days 0 and 7. Cell suspensions and conditioned media were analyzed for DNA and secretion of PRG4, respectively. Flow cytometric histograms were deconvolved so that the number of cells in each doubling generation contributing to the final cell population could be estimated. Chondrocytes were consistently and intensely labeled with PKH26 through 7 cycles of division. At day 7 of culture, >97% of superficial zone cells seeded at low or high density could be distinguished as fluorescent, as could middle zone cells seeded at high density. Retention of cell fluorescence after PKH26 labeling and lack of adverse effects on cell proliferation and synthesis of PRG4 suggest that PKH26 can be useful in determining the fate and function of implanted chondrocytes in vivo, as well as monitoring proliferation in vitro. PMID- 16715533 TI - Anaerobic fermentation of glycerol by Escherichia coli: a new platform for metabolic engineering. AB - The worldwide surplus of glycerol generated as inevitable byproduct of biodiesel fuel and oleochemical production is resulting in the shutdown of traditional glycerol-producing/refining plants and new applications are needed for this now abundant carbon source. In this article we report our finding that Escherichia coli can ferment glycerol in a pH-dependent manner. We hypothesize that glycerol fermentation is linked to the availability of CO(2), which under acidic conditions is produced by the oxidation of formate by the enzyme formate hydrogen lyase (FHL). In agreement with this hypothesis, glycerol fermentation was severely impaired by blocking the activity of FHL. We demonstrated that, unlike CO(2), hydrogen (the other product of FHL-mediated formate oxidation) had a negative impact on cell growth and glycerol fermentation. In addition, supplementation of the medium with CO(2) partially restored the ability of an FHL deficient strain to ferment glycerol. High pH resulted in low CO(2) generation (low activity of FHL) and availability (most CO(2) is converted to bicarbonate), and consequently very inefficient fermentation of glycerol. Most of the fermented glycerol was recovered in the reduced compounds ethanol and succinate (93% of the product mixture), which reflects the highly reduced state of glycerol and confirms the fermentative nature of this process. Since glycerol is a cheap, abundant, and highly reduced carbon source, our findings should enable the development of an E. coli-based platform for the anaerobic production of reduced chemicals from glycerol at yields higher than those obtained from common sugars, such as glucose. PMID- 16715534 TI - Modeling the partial nitrification in sequencing batch reactor for biomass adapted to high ammonia concentrations. AB - Partial nitrification has proven to be an economic way for treatment of industrial N-rich effluent, reducing oxygen and external COD requirements during nitrification/denitrification process. One of the key issues of this system is the intermediate nitrite accumulation stability. This work presents a control strategy and a modeling tool for maintaining nitrite build-up. Partial nitrification process has been carried out in a sequencing batch reactor at 30 degrees C, maintaining strong changing ammonia concentration in the reactor (sequencing feed). Stable nitrite accumulation has been obtained with the help of an on-line oxygen uptake rate (OUR)-based control system, with removal rate of 2 kg NH4 (+)-N x m(-3)/day and 90%-95% of conversion of ammonium into nitrite. A mathematical model, identified through the occurring biological reactions, is proposed to optimize the process (preventing nitrate production). Most of the kinetic parameters have been estimated from specific respirometric tests on biomass and validated on pilot-scale experiments of one-cycle duration. Comparison of dynamic data at different pH confirms that NH3 and NO2- should be considered as the true substrate of nitritation and nitratation, respectively. The proposed model represents major features: the inhibition of ammonia-oxidizing bacteria by its substrate (NH3) and product (HNO2), the inhibition of nitrite oxidizing bacteria by free ammonia (NH3), the INFluence of pH. It appears that the model correctly describes the short-term dynamics of nitrogenous compounds in SBR, when both ammonia oxidizers and nitrite oxidizers are present and active in the reactor. The model proposed represents a useful tool for process design and optimization. PMID- 16715535 TI - Prenatal diagnosis of free sialic acid storage disorders (SASD). AB - Free sialic acid storage disorders, Salla disease (SD) and Infantile sialic acid storage disease (ISSD), are lysosomal storage diseases due to impaired function of a sialic acid transporter, sialin, at the lysosomal membrane. Several mutations of the sialin gene, SLC17A5, are known, leading either to the severe neonatal/infantile disease or to the milder, adult-type developmental disorder, Salla disease. Free sialic acid accumulation in lysosomes causes increased tissue concentration and consequently elevated urinary excretion. Prenatal diagnosis of SASD is possible either by determination of free sialic acid concentration or by mutation analysis of the SLC17A5 gene in fetal specimen, in chorionic villus biopsy particularly. Both techniques have been successfully applied in several cases, sialic acid assay more often in ISSD cases but mutation analysis preferentially in SD. Sialic acid assay of amniotic fluid supernatant or cultured amniotic fluid cells may give erroneous results and should not be used for prenatal diagnosis of these disorders. The present comments are mainly based on our experience of prenatal diagnosis of SD in Finnish families. A founder mutation in SLC17A5 gene, 115C-> T, represents 95% of the disease alleles in the Finnish SD patients, which provides a unique possibility to apply mutation analysis. Therefore, molecular studies have successfully been used in 17 families since the identification of the gene and the characterization of the SD mutations. Earlier, eight prenatal studies were performed by measuring the free sialic acid concentration in chorionic villus samples. PMID- 16715536 TI - Antidepressant prevalence for youths: a multi-national comparison. AB - OBJECTIVE: To compare antidepressant prevalence data in youths across three western European countries (Denmark, Germany, and the Netherlands) with US regional data in terms of age and gender and to show proportional subclass antidepressant (ATD) use. METHOD: A population-based analysis of administrative claims data for the year 2000 was undertaken in 0 to 19-year-old enrollees who were part of the insured populations from four countries having a total of from 72,570 to 480,680 members. RESULTS: ATD medication utilization in the US dataset (1.63%) exceeded that of three Western European countries (prevalence ranged from 0.11 to 0.54%) by at least 3-fold. There were major variations in the use of subclasses: tricyclic antidepressants (TCAs) predominated in Germany while selective serotonin reuptake inhibitors (SSRIs) predominated in the US, Denmark and the Netherlands. CONCLUSIONS: Cross-national variations should be further explored to understand the factors related to these differences and how prevalence differences relate to effectiveness and safety. Community-based cohorts should be followed to establish outcomes in the usual practice setting. PMID- 16715537 TI - Prenatal diagnosis of Freeman-Sheldon syndrome and usefulness of an ultrasound fetal lip width normogram. AB - OBJECTIVES: Freeman-Sheldon syndrome (FSS) is a rare genetic condition with facial and joint abnormalities. Here we report the first case of prenatal diagnosis in a fetus with negative familial history for this syndrome. A chart of the fetal mouth length was generated and its usefulness verified. METHODS: A cross-sectional study was carried out to derive standard charts for fetal lip lengths. The study population was made up of 371 fetuses between 15 and 38 weeks of gestation examined only once for this purpose. A statistical analysis previously recommended to derive normal ranges in fetal growth charts was employed. RESULTS: A 34-year-old woman was referred at 28 weeks of gestation for polyhydramnios and clubfeet. A detailed scan confirmed those findings and showed joint contractures of hands, a severe microstomia with a 'whistling' mouth, and an H-shaped mark over the chin. FSS was suspected, though there was no family history for this genetic syndrome. After emergency delivery at 32 weeks, the syndrome was confirmed. The infant died two months later from pneumonia. Upon construction of a standard mouth length chart, it was found that the fetal lip width in this case of FSS was well below the 5th centile. CONCLUSION: Here we present a standard chart for fetal lip width. We demonstrated its usefulness in cases with fetal face anomalies. Specific charts are fundamental for an objective assessment of the fetal parameters. PMID- 16715538 TI - Prenatal diagnostic indicators of paternal uniparental disomy 14. AB - OBJECTIVES: To present clinical findings of a child with paternal uniparental isodisomy 14 (pat UPD14) focusing on relevant prenatal characteristics. METHODS/RESULTS: Ultrasonography at 23 weeks of gestation of a 37-year-old multigravid woman revealed a fetus with polyhydramnios, small thorax, and short, distinctively angled ribs. Fetal karyotype was 46,XY. The child was born spontaneously at 35 weeks with poor neonatal adaptation. From birth, he presented with severe respiratory insufficiency due to severe thoracic malformation. Clinical examination revealed a small, bell-shaped thorax, redundant lax skin, mild contractures of the fingers and dysmorphic facial features. Chest X rays showed short, abnormally curved ribs that suggested the possibility of pat UPD14, which was confirmed by molecular analysis. CONCLUSION: Pat UPD14 is associated with a distinct clinical phenotype. Prognosis is poor because of severe respiratory insufficiency and neurodevelopmental retardation. Our report confirms salient postnatal signs of previous descriptions, especially the characteristic radiological abnormalities with ribs showing a 'coat-hanger' configuration. Retrospective fetal ultrasound of our case allowed the identification of this pathognomonic feature prenatally, which makes it possible to consider pat UPD14 at routine prenatal sonography, in particular in combination with a small bell shaped thorax and polyhydramnios. PMID- 16715539 TI - No association between inhaled corticosteroids and whole body DXA in postmenopausal women. AB - PURPOSE: Postmenopausal women treated with corticosteroids are regarded as a high risk group due to the effect of both natural bone loss and possible adverse effects of treatment with inhaled corticosteroids (IC). OBJECTIVE: To compare bone mineral density (BMD) in postmenopausal women exposed only to IC (IC group, n = 106) with that of BMD in women not exposed to corticosteroids (n = 124) and women exposed to oral and/or intra-articular injections in addition to inhaled corticosteroids (OC group, n = 31). The women were recruited from a population based prospective cohort study. METHODS: Dual X-ray absorptiometry (DXA) technique was used to measure BMD in whole body, spine, pelvis and lower extremities. A health questionnaire and an interview about past and present medication use were used. RESULTS: The mean duration and dose of IC were 9.5 +/- 4.5 years and 615 microg daily. Whole body BMD did not significantly differ between the IC group (1.103 g/cm(2)) and the unexposed group (1.087 g/cm(2)). Within the IC group, BMD stratified for cumulative dose of IC, duration or current dose above or below 800 microg did not differ. Z-score BMD for tertiles did not differ when comparing the IC and OC groups. CONCLUSION: No difference in BMD was noted between postmenopausal women exposed to inhaled corticosteroids and unexposed controls nor was there any dose response relationship between inhaled corticosteroid therapy and BMD. PMID- 16715540 TI - Target-assisted selection of galactosyltransferase binders from dynamic combinatorial libraries. An unexpected solution with restricted amounts of the enzyme. PMID- 16715541 TI - Visual artificial tongue for quantitative metal-cation analysis by an off-the shelf dye array. AB - A chemical-probe array composed of 47 off-the-shelf dyes was prepared in solution format (New York Tongue 1: NYT-1) and was tested in the identification and quantitation of 47 cation analytes, including 44 metal ions, in addition to H(+), NH(4) (+), and tetrabutylammonium (TBA). The cation solutions were tested in a series of concentrations and the fold-change in effective absorbance was analyzed by principal-component analysis (PCA), hierarchical-cluster analysis (HCA), and nearest-neighbor decision to determine both identity and quantity of the analytes. Apart from alkali-metal ions (Na(+), K(+), Li(+), Cs(+), and Rb(+)), which behave very similarly to each other due mainly to their low response, most of the cations were clearly distinguishable at 10 mM concentration. The practical detection limit of each analyte was also determined by a sequential dilution and the nearest-neighbor decision method. In the finalized working analyte concentration range (approximately 10 mM down to 0.33 microM), by considering alkali metals as one analyte group, most of the analytes were correctly identified (99.4 %). Furthermore, the success rate at which the concentration of each analyte was correctly determined was also high (96.8 %). PMID- 16715542 TI - Amplification and diversity analysis of ketosynthase domains of putative polyketide synthase genes in Aspergillus ochraceus and Aspergillus carbonarius producers of ochratoxin A. AB - The diversity of polyketide synthase (PKS) genes in Aspergillus ochraceus NRRL 3174 and Aspergillus carbonarius 2Mu134 has been investigated using different primer pairs previously developed for the ketosynthase (KS) domain of fungal PKSs. Nine different KS domain sequences in A. ochraceus NRRL 3174 as well as five different KS domain sequences in A. carbonarius 2Mu134 have been identified. The identified KS fragments were distributed in five different clusters on the phylogenetic tree, indicating that they most probably represent PKSs responsible for different functions. PMID- 16715543 TI - Microbiologicals for deactivating mycotoxins. AB - Mycotoxins are secondary metabolites of fungi affecting human and animal health. Five classes of mycotoxins are of major concern in animal husbandry, namely aflatoxins, trichothecenes, zearalenone, ochratoxins, and fumonisins. Due to their diverse structure these fungal toxins are able to cause a great variety of acute symptoms in animals. Clay minerals have been used in animal nutrition to bind mycotoxins, but the binders are only very specific for aflatoxins but not for other toxins. A novel strategy to control the problem of mycotoxicoses in animals is the application of microorganisms capable of biotransforming mycotoxins into nontoxic metabolites. The microbes act in the intestinal tract of animals prior to the resorption of the mycotoxins. A Eubacterium (BBSH 797) strain is able to deactivate trichothecenes by reduction of the epoxide ring (CAST, Mycotoxins, Risks in Plant, Animal and Human Systems, Task Force Report 139, Council of Agricultural Science and Technology, Ames Iowa 2003, p. 10.; Binder, E. M., Binder, J., Ellend, N., Schaffer, E. et al., in: Miraglia, M., van Egmond, H., Brera, C., Gilbert, J. (Eds.), Mycotoxins and Phycotoxins- Developments in Chemistry, Toxicology and Food Safety, Alaken, Fort Collins 1996, pp. 279-285). This strain was isolated out of bovine rumen fluid and the mode of action was proven in vitro and also in vivo. Further a novel yeast strain, capable of degrading ochratoxin A and zearalenone was isolated and characterized (Bruinink, A., Rasonyi, T., Sidler, C., Nat. Toxins 1999, 6, 173-177; Schatzmayr, G., Heidler, D., Fuchs, E., Mohnl, M. et al., Mycotoxin Res. 2003, 19, 124-128.) Due to the yeasts affiliation to the genus of Trichosporon and its property to degrade mycotoxins this strain was named Trichosporon mycotoxinivorans (Trichosporon MTV, 115). PMID- 16715544 TI - Balkan endemic nephropathy: role of ochratoxins A through biomarkers. AB - Several studies implicated mycotoxins, in endemic kidney disease geographically limited to Balkan region (Balkan endemic nephropathy (BEN)). In Bulgaria, much higher prevalence of ochratoxin A (OTA), exceeding 2 microg/L, was observed in the blood of affected population. OTA is found more often in the urine of people living in BEN-endemic villages. To confirm and quantify exposure to OTA in Vratza district, we followed up OTA intake for 1 month, OTA in blood and urine from healthy (20-30 years old) volunteers, from two villages with high risk for BEN disease. Food samples were collected daily, blood and urine at the beginning of each week. Relations between increasing OTA intake, blood concentration and elimination of OTA in urine have been studied in rats. Average weekly intake of OTA varies from 1.9 to 206 ng/kg body weight, twice tolerable weekly intake recommended by JECFA. OTA blood concentrations are in the same range as previously reported in this region with concentrations reaching 10 microg/L. Weekly OTA food intake is not directly correlated with blood and urine concentrations. Biomarkers of biological effects such as DNA adducts were detected in patients affected by urinary tract tumours (UTT) and in rat study. All these plead for the implication of OTA, in BEN and UTT. PMID- 16715545 TI - Characterization of filamentous fungi isolated from Moroccan olive and olive cake: toxinogenic potential of Aspergillus strains. AB - During the 2003 and 2004 olive oil production campaigns in Morocco, 136 samples from spoiled olive and olive cake were analyzed and 285 strains were isolated in pure culture. Strains included 167 mesophilic strains belonging to ten genera: Penicillium, Aspergillus, Geotrichum, Mucor, Rhizopus, Trichoderma, Alternaria, Acremonium, Humicola, Ulocladium as well as 118 thermophilic strains isolated in 2003 and 2004, mainly belonging to six species: Aspergillus fumigatus, Paecilomyces variotii, Mucor pusillus, Thermomyces lanuginosus, Humicola grisea, and Thermoascus aurantiacus. Penicillium and Aspergillus, respectively, 32.3 and 26.9% of total isolates represented the majority of mesophilic fungi isolated. When considering total strains (including thermotolerant strains) Aspergillus were the predominant strains isolated; follow-up studies on mycotoxins therefore focused primarily on aflatoxins (AFs) and ochratoxin A (OTA) from the latter strains. All isolated Aspergillus flavus strains (9) and Aspergillus niger strains (36) were studied in order to evaluate their capacity to produce AFs and OTA, respectively, when grown on starch-based culture media. Seven of the nine tested A. flavus strains isolated from olive and olive cake produced AF B1 at concentrations between 48 and 95 microg/kg of dry rice weight. As for the A. niger strains, 27 of the 36 strains produced OTA. PMID- 16715546 TI - Sampling: the weak link in the sanitary quality control system of agricultural products. AB - To ensure a high level of consumer protection, the European Union has in the past years published several regulations setting very low limits for a given number of food contaminants (pesticides, mycotoxins, heavy metals) in many agricultural products (cereals, oilseeds, dry fruits, coffee, spices, etc). These new regulations regarding the sanitary quality of agricultural products, compel both economic operators and officials of different EU member states to set up sampling plans and rigorous analyses aimed at checking whether a product lot complies with the required standards prior to its release on the market. While the laboratory analysis management today is outstanding thanks to the validated and efficient detection methods and procedures available for quality assurance in laboratories (accreditation), this is not necessarily true of the sampling operation, which seems to be the weak link in the sanitary control system for agricultural products. The sampling operation is often the main source of error when assessing the sanitary quality of a lot of agricultural commodities, with both commercial (downgrading of the product) and sanitary (marketing of a product which poses a health risk for the consumer) consequences. Therefore, it is essential for the operators involved to be aware of the significance and difficulties of the sampling operation, which requires important equipment and human resources. Furthermore, drawing up specific standards and guidelines, as well as setting up quality assurance procedures, at the level in charge of carrying out this delicate and important operation, are necessary. PMID- 16715547 TI - Welfare in Lancashire cotton mills in the 1940s. PMID- 16715548 TI - Obesity, smoking, and psoriasis. PMID- 16715549 TI - APCcdh1 activity in mouse oocytes prevents entry into the first meiotic division. AB - Fully grown mammalian oocytes maintain a prophase I germinal-vesicle stage arrest in the ovary for extended periods before a luteinizing hormone surge induces entry into the first meiotic division. Cdh1 is an activator of the anaphase promoting complex (APC) and APCcdh1 is normally restricted to late M to early G1 phases of the cell cycle. Here, we find that APCcdh1 is active in mouse oocytes and is necessary to maintain prophase arrest. PMID- 16715551 TI - Sociodemographic associations of physical, emotional, and sexual intimate partner violence in Spanish women. AB - PURPOSE: To measure the lifetime prevalence of physical, emotional, and sexual Intimate Partner Violence (IPV) in women attending general practice in Spain and to assess sociodemographic factors associated with the different types of abuse. METHODS: The sample included 1402 randomly selected women, aged 18 to 65 years, attending general practice in three Spanish regions. A self-administered structured questionnaire specifically developed for the study was used. Given the overlap between the different types of lifetime abuse, various categories were created by combining the different types. Multiple logistic regression models were developed to identify the sociodemographic factors independently associated with the different categories of abuse. RESULTS: Lifetime prevalence of IPV was 32%, and 14.4% referred having experienced emotional abuse alone. Factors independently associated with all categories of abuse included being separated/divorced/widowed, having three or more children, and lacking social support. Increasing age was protective for physical and emotional abuse (odds ratio = 0.96; 95% confidence interval = 0.93-0.99). Women with the lowest monthly family income (< 600 euro) (1 euro = 1.32 euro) were most likely to experience all IPV categories, except for emotional abuse alone. CONCLUSIONS: IPV is a common problem in general practice. The identification of factors associated with this public health problem is essential for its prevention and treatment. PMID- 16715552 TI - Long-term care: long-term care insurance--2005. End of Year Issue Brief. AB - As the "Baby Boom" generation approaches retirement, state and federal lawmakers are struggling to ensure that the nation's long-term care system will provide adequate services for the growing number of senior citizens. A 2003 Administration on Aging report predicted that the elderly population will double by 2030. Accordingly, policymakers must prepare for the impending squeeze on public health and Medicaid resources. Many consumers are exploring private long term care insurance options as a means of preparing for the cost of eldercare. Yet, a lack of market uniformity has rendered the long-term care insurance industry somewhat difficult for consumers to decipher. In addition, senior care insurance is often costly, particularly for those over age 50. PMID- 16715553 TI - NF-kappaB: a signal for cancer. PMID- 16715554 TI - Public health: tobacco taxes and Internet sales--2005. End of Year Issue Brief. AB - Raising tobacco taxes is an action that resonates with lawmakers, public health and anti-tobacco advocates, and the majority of the electorate. The relatively broad base of support for increasing excise taxes and the potential for increased tax revenue mitigate the concerns over targeting tobacco-users--23 percent of the population--to pay for state programs and the unreliability of the tobacco tax as a permanent source of revenue. Tobacco excise taxes generated $10.2 billion, or about 1.5 percent of all states' revenue. Characterized as sin taxes or user fees and viewed as an effective method to deter price-sensitive adolescents from using tobacco, excise taxes on tobacco have increased in an unprecedented number of states since November 2001. Previously, no more than three states, on average, had increased cigarette taxes in a year. The legislative action is viewed as a politically safe and relatively easy way to raise taxes and increase revenue without incurring the wrath of anti-tax voters. During this period the increases in tobacco taxes ranged from a $0.12 per pack increase in Louisiana to a $0.75 increase in Massachusetts and Michigan. Nationwide the state tax rate per pack of cigarettes ranges from a low in South Carolina of $0.07 to $2.46 in Rhode Island. The mean tobacco excise tax for the nation is approximately $0.92. With an excise tax increase, states can channel needed funds to programs favored by voters in economically strapped times. Indeed, many of the 44 states that increased their tobacco tax announced that the revenue would permit the state to restore or at least reduce proposed cuts to Medicaid and other health programs. Excise taxes also place little administrative burden on states, since the wholesaler pays the tax directly to the state and the additional cost then is passed on to the consumer. PMID- 16715555 TI - TX: agency nurse falls on wet hospital floor: failure to plead comp. as 'exclusive remedy'. PMID- 16715556 TI - [Platelet satellitism]. PMID- 16715557 TI - Redefining radiology education for first-year medical students: shifting from a passive to an active case-based approach. AB - RATIONALE AND OBJECTIVES: The aim of the study is to design and implement an elective course in which first-year medical students learn about radiology by actively integrating imaging with hospital records and radiology reports, rather than passively observing radiologists or images. MATERIALS AND METHODS: Twenty students enrolled in an elective entitled "Introduction to Clinical Radiology." Students were divided into groups of two or three and given a case file describing the hospital visit of a patient with a common computed tomographic (CT) diagnosis. Files contained a PowerPoint presentation with all images from the patient's CT examination (allowing students to scroll through the images as if viewed on a picture archive and communication system workstation), as well as emergency department records, admission and discharge summaries, surgical notes, pathology reports, and radiology reports. Group discussion was facilitated by radiology staff acting as consultants, rather than instructors. As a final project, each group presented its case to the students and faculty. RESULTS: Compared with before the course, students' understanding of the role of radiologists in the clinical management of patients significantly increased, but interest in radiology as a specialty was unchanged. Students significantly preferred scrolling through images on their own, rather than being shown static images in a didactic format. Students significantly agreed that they would recommend the course to a classmate. CONCLUSION: Case-based instruction in radiology, in which first-year medical students are actively engaged in the integration of clinical and imaging information, can affect students' views on the role of radiologists and their clinical management of patients. PMID- 16715558 TI - The ethics of human gene therapy. PMID- 16715559 TI - Toward regulating genetic engineering. PMID- 16715560 TI - Does bioethics provide answers?: Secular and religious bioethics and our procreative future. PMID- 16715561 TI - Method, mediations, and the moral dimensions of preimplantation genetic diagnosis. PMID- 16715562 TI - Catholic bioethics and the case of Terri Schiavo. PMID- 16715563 TI - Shattering the neutral surrogate myth in end-of-life decisionmaking: Terri Schiavo and her family. PMID- 16715564 TI - Why human beings may use animals. PMID- 16715565 TI - The limited regulatory potential of medical technology assessment. PMID- 16715566 TI - Does assessment of medical practices have a future? PMID- 16715567 TI - Technology assessment and the doctor-patient relationship. PMID- 16715568 TI - A wrongful birth? PMID- 16715569 TI - Pro-life nation: what happens when you completely criminalize abortion? Over the last eight years, El Salvador has found out. PMID- 16715570 TI - Taking the least of you: most of us have tissue or blood samples on file somewhere, whether we know it or not. What we don't typically know is what research they are being used for or how much money is being made from them. And science may want to keep things that way. PMID- 16715571 TI - The other stem-cell debate: to test the potential curative powers of human embryonic stem cells, biologists want to inject them into lab animals. Creating such chimeras makes perfect sense, to a point: a sheep with a human liver? O.K. A mouse brain made up of human cells? Maybe. But a chimp that sobs? PMID- 16715572 TI - The other mother: K. provided the eggs, her lover the womb, and for nearly six years the two women raised the twin girls thus conceived. But when the women broke up, K. learned how fragile the definition of motherhood could be. PMID- 16715573 TI - Uncharitable? Nonprofit hospitals, with tax exemptions for serving the poor, are quietly charging indigent patients--and even suing over unpaid bills. How did an arrangement to serve the needy become so unhealthy? PMID- 16715574 TI - Curing Casanova: a pharmacological infidelity inhibitor just may be on its way. Should we welcome it? PMID- 16715575 TI - [A young man with acute dilated cardiomyopathy]. PMID- 16715576 TI - Atypical enteropathogenic Escherichia coli: typical pathogens? PMID- 16715577 TI - Therapy in type 2 diabetes: insulin glargine vs. NPH insulin both in combination with glimepiride. AB - BACKGROUND: Type 2 diabetes (T2DM) patients often fail to achieve adequate glycemic control with oral antidiabetic drugs (OADs). Insulin has been shown to improve glycemic control in these patients but with increased risk of hypoglycemia. This study compared the efficacy and safety of insulin glargine and NPH insulin, both in combination with a once-daily fixed dose of glimepiride, in terms of glycemic control and incidence of hypoglycemia. METHODS: In this open label, 24-week randomized trial in ten Latin American countries, T2DM patients poorly controlled on OADs (HbA1c > or = 7.5 and < or = 10.5%) received glimepiride plus insulin glargine (n = 231) or NPH insulin (n = 250) using a forced titration algorithm. The primary endpoint was the equivalence of 24-week mean changes in HbA1c. RESULTS: Insulin glargine and NPH insulin achieved similar HbA1c reductions (adjusted mean difference -0.047; 90% CI -0.232, 0.138; per protocol analysis). Confirmed nocturnal hypoglycemia was significantly lower with insulin glargine vs. NPH insulin (16.9 vs. 30.0%; p <0.01; safety analysis). Patients receiving insulin glargine were significantly more likely to achieve HbA1c levels < 7.0% without hypoglycemia (27 vs. 17%; p = 0.014; per-protocol analysis). There was a more pronounced treatment satisfaction improvement with insulin glargine vs. NPH insulin (p <0.02; full analysis). The proportion of patients who lost time from work or normal activities due to diabetes was lower with insulin glargine vs. NPH (1.8 vs. 3.3%; full analysis). CONCLUSIONS: In patients with T2DM, inadequately controlled on OADs, once-daily insulin glargine plus glimepiride is effective in improving metabolic control with a reduced incidence of nocturnal hypoglycemia compared with NPH insulin. PMID- 16715578 TI - Computer-assisted telephone interview techniques. PMID- 16715579 TI - Lack of transmission of vaccinia virus. PMID- 16715580 TI - [Drug-induced cardiomyopathy]. PMID- 16715581 TI - Discrimination between highly pathogenic and low pathogenic H5 avian influenza A viruses. PMID- 16715583 TI - HIV-1 CRF07_BC infections, injecting drug users, Taiwan. PMID- 16715582 TI - Rift Valley fever in goats, Cameroon. PMID- 16715584 TI - Henipavirus in Pteropus vampyrus bats, Indonesia. PMID- 16715585 TI - Chlamydialike organisms and atherosclerosis. PMID- 16715586 TI - Maculopathy in dengue fever. PMID- 16715587 TI - Pulmonary tuberculosis and SARS, China. PMID- 16715588 TI - Tetanus in injecting drug users, United Kingdom. PMID- 16715589 TI - Xanthine oxidase inhibiting flavonol glycoside from Amberboa ramosa. AB - A new flavonol glycoside (1) has been isolated from the ethyl acetate soluble fraction of Amberboa ramosa and assigned the structure 5,7,4'-trihydroxy-3,8 dimethoxylflavone 5-O-beta-D-gluco-pyranoside (1). In addition, 6,4'-dihydroxy 3,5,7-trimethoxyflavone (2), 5,7-dihydroxy-4'-methoxyflavone (3) and (23R)-5alpha cycloart-24-ene-3beta,21,23-triol (4) have also been reported for the first time from this species. The structures were deduced on the basis of 1D and 2D NMR techniques. The compounds 1-3 displayed weak to moderate inhibition against the xanthine oxidase enzyme. PMID- 16715590 TI - Manifesting ecologic and microbial connections. PMID- 16715591 TI - Edinburgh's 'occasional auditors'. PMID- 16715592 TI - More data on hormone therapy and coronary artery disease: comments on recent publications from the WHI and Nurses' Study. PMID- 16715593 TI - Chronic disease management systems registries in rural health care. AB - Health care quality is being addressed from a variety of policy perspectives. The 2001 Institute of Medicine report, Crossing the Quality Chasm, calls for sweeping action involving a five-part strategy for change in the U.S. health care system. This agenda for change includes use of evidence-based approaches to address common conditions, the majority of which are chronic. A Chronic Disease Management System (CDMS), or registry, is a tool that helps providers efficiently collect and analyze patient information to promote quality care for the rural population. CDMSs can provide a technological entry point for the impending use of Electronic Medical Records. A CDMS is a patient-centered electronic database tool that helps providers diagnose, treat, and manage chronic diseases. The purpose of this brief is to discuss the different types of CDMSs used by a sample of 14 state organizations and 19 local rural clinics in Maine, Nebraska, New Mexico, South Carolina, Washington, and Wisconsin. As part of a larger study examining the challenges and innovations in implementing disease management programs in rural areas, we conducted interviews with national, state, and local contacts. During interviews, respondents helped us understand the usefulness and functionalities of commonly used CDMSs in rural facilities. Our focus was on the use of CDMSs in the management of diabetes, a disease prevalent in rural populations. KEY FINDINGS: (1) CDMSs are readily available to rural clinics and are being implemented and maintained by clinic staff with minimal expenditures for technology. (2) Use of a standardized system in a collaborative helps provide data comparisons and share costs involved with technical assistance services across the group. PMID- 16715594 TI - Language barriers pose a risk for California HMO enrollees. PMID- 16715596 TI - Cytomegalovirus infection in lung transplant patients: the role of prophylaxis and recipient-donor serotype matching. AB - Cytomegalovirus (CMV) remains an important cause of morbidity and mortality in lung transplant recipients. We investigated the incidence of CMV infection in relation to CMV prophylaxis, and recipient-donor CMV serotype, in a cohort of 250 consecutive lung transplant recipients. All patients received 3 months CMV prophylaxis with acyclovir (n = 67) or gancyclovir (n = 183). Recipient-donor CMV serotype matching was performed in patients receiving acyclovir: R+/D+(n = 38), R+/D-(n = 10), R-/D+(n = 1), R- /D-(n = 16), unknown (n = 2). Recipient-donor CMV serotype matching was not performed in patients receiving gancyclovir: R+/D+(n = 71), R+/D-(n = 42), R-/D+(n = 38), R-/D-(n = 31), unknown (n = 1). The overall incidence of CMV infection was 51% (n = 34) in the acyclovir group, and 42% (n = 77) in the gancyclovir group (p = 0.14). During the first 9 months after transplantation, the rate of CMV infection was higher in the acyclovir group (42%) compared with the gancyclovir group (30%) (p = 0.005). Multivariate analysis demonstrated the incidence of CMV infection during the first 9 months was higher for acyclovir prophylaxis (p<0.001) and R-/D+ serostatus (p<0.001) and lower with R-/D- serostatus (p = 0.02). In conclusion, gancyclovir significantly delays the onset of first CMV infection among lung transplant patients. CMV surveillance and choice of prophylaxis may be modified according to donor recipient CMV serotype. PMID- 16715597 TI - Introduction: addressing cardiometabolic risk. PMID- 16715595 TI - A binational cohort study of ventilator-associated pneumonia in Denmark and Australia. AB - The objective was to determine the incidence and prognosis of ventilator associated pneumonia (VAP) in intensive care units (ICUs) in Melbourne (29-bed ICU), Australia and Aarhus and Aalborg (22-bed unit and 8-bed ICU, respectively), Denmark and to characterize participating ICUs with regard to prevalence of nosocomial type bacterial pathogens, antibiotic resistance and antibiotic consumption. In this prospective cohort study 25 patients in Melbourne and 32 patients in Aarhus + Aalborg had a first episode of VAP. The incidence of VAP per 1000 ventilator d was 6.2 in Melbourne and 9.5 in Aarhus + Aalborg. Case fatality during hospital admission was 28% and 59%, respectively (unadjusted odds ratio (OR) 0.3, 95% confidence interval (CI) 0.1-0.8). OR adjusted for age and APACHE II score was 0.2 (95% CI 0.1-1.0). Nosocomial type pathogens including methicillin resistant Staphylococcus aureus were more prevalent in Melbourne, and antibiotic consumption per VAP patient was 35% higher in Melbourne than in Aarhus + Aalborg. To judge from the present data, there seems to be a complicated interrelationship between prognosis on the 1 hand and antibiotic consumption and resistance on the other. A more favourable prognosis was found in Melbourne, where levels of antibiotic consumption and antimicrobial resistance were higher than in Aarhus + Aalborg. PMID- 16715598 TI - Which is worse: underweight or overweight. PMID- 16715599 TI - Eye safety is everyone's business. PMID- 16715600 TI - [From fundamentals to clinical application of dose area product meter]. PMID- 16715601 TI - [The point of clinical cardiac MRI]. PMID- 16715602 TI - [Why do we need the systemization of the medicine and how do we analyze the data?]. PMID- 16715603 TI - [How should we accept the risk from low level radiation?]. PMID- 16715604 TI - [International comparison of dosimetry calibration coefficients for absorbed dose in high energy photon beams and electrons]. PMID- 16715605 TI - [Quality assurance (QA) guideline for medical imaging display systems]. PMID- 16715606 TI - New report synthesizes transformation recommendations. PMID- 16715607 TI - Key factors impacting the delivery and financing of oncology care in the United States. PMID- 16715608 TI - Exploring the ontology of space, place, and meaning. PMID- 16715609 TI - Aprotinin in the management of coagulopathy associated with amniotic fluid embolus. AB - Amniotic fluid embolus, also known as anaphylactoid syndrome of pregnancy is a rare complication of pregnancy. When it occurs, the maternal mortality rate may be as high as 86%, and in survivors, the morbidity rate may be just as high. Hallmark clinical features include maternal cardiovascular collapse with disseminated intravascular coagulation, and fetal distress. Management centers on strategies to improve oxygenation, support circulation, and correct the coagulopathy. We report the case of a patient who developed amniotic fluid embolus and was effectively managed with aprotinin to control the coagulopathy associated with this devastating complication of pregnancy. PMID- 16715610 TI - Pharmacokinetics of daptomycin in a critically ill adolescent with vancomycin resistant enterococcal endocarditis. AB - Daptomycin is a lipopeptide antibiotic active against multidrug-resistant gram positive organisms. Our search of the literature found no published pediatric pharmacokinetic data. We report the use of pharmacokinetic analysis of daptomycin in a 13-year-old boy with vancomycin-resistant Enterococcus faecium endocarditis. Pharamcokinetic parameters were found to be significantly different from published adult parameters, such as a faster elimination rate, shorter half-life, and increased clearance. These age-related differences in the pharmacokinetic profile of daptomycin have significant dosing implications. As the use of this drug for off-label indications and in pediatric populations increases, it is important for clinicians to better understand the drug's pharmacokinetic profile in these patient populations. PMID- 16715611 TI - Tsunami induced hyperglycemia and diabetes mortality--two studies from south India. PMID- 16715612 TI - Stress and undetected hyperglycemia in southern Indian coastal population affected by tsunami. AB - AIM: Natural calamities are known to result in higher stress conditions and also result in adverse health outcomes including development of non-communicable diseases. The impact of tsunami on mental stress and prevalence of hyperglycemic conditions was assessed in a population affected by the calamity in coastal populations of southern India. METHODS: Two populations similar in demography and physical characteristics, but, one affected by tsunami were selected for a comparative study. Subjects aged 20 years or above were randomly selected (control population n = 1176; tsunami population n = 1184). Details of stress were assessed using Harvard trauma questionnaire and scores were assigned. Glucose tolerance was assessed using 2h capillary blood glucose (75gms glucose load) and diagnosis was made using WHO criteria. RESULTS: Stress score was significantly higher in tsunami population. Although the total prevalence of diabetes was similar (control - 10.0 %; tsunami population - 10.5 %) prevalence of undetected diabetes (5.7 % vs. 3.8 %; Z = 9.54, P < 0.001) and impaired glucose tolerance (9.8 % vs. 8.3 %; Z = 12.83, P < 0.001) higher in the tsunami area. Stress score was higher in women and in the young in the tsunami area. CONCLUSION: Population affected by tsunami was under high stress and also showed a high prevalence of undetected diabetes and impaired glucose tolerance. PMID- 16715614 TI - Evaluation of enzymes in pyogenic and tuberculous meningitis. AB - OBJECTIVES: Lactic dehydrogenase (LDH), creatine kinase (CK) and gamma glutamyl transpeptidase (GGTP) were measured serially in cerebrospinal fluid (CSF) and serum in twenty five cases of meningitis and an equal number of age and sex matched healthy control subjects with an aim to find out their diagnostic and prognostic significance in cases of meningitis. METHODS: The enzymatic activity was measured serially (day 0, 4th and 7th) in cerebrospinal fluid (CSF) and serum in twenty-five cases of meningitis consisting of fifteen cases of pyogenic meningitis (PM) and ten of tuberculous meningitis (TBM) and an equal number of age and sex matched healthy control. The clinical details including the level of consciousness and neurological deficit were correlated with the enzymatic activity and prognosis. RESULTS: The levels of these enzymes were significantly elevated in all the cases of meningitis in serum as well as CSF as compared to control subjects. The activity was significantly higher in pyogenic than tuberculous meningitis (p<0.001) and it was higher in CSF than in serum (p<0.001). The maximum elevation in activity of GGTP and LDH were seen on the first day whereas CK was highest on 4th day and thereafter, the activity of all the enzymes declined in the majority of cases who had shown clinical improvement. However, in three cases of pyogenic and five cases of tuberculous meningitis, the enzymatic activity on subsequent estimation, increased serially. All these eight cases died. Further, the basal enzymatic activity in all these eight cases that died was higher as compared to those who survived. Of all the enzymes, CSF GGTP levels correlated best with the clinical picture. CONCLUSIONS: It is concluded that GGTP, CK and LDH were significantly elevated in cases of meningitis. It was not possible to differentiate the type of meningitis on the basis of enzymatic activity in any of them. However, it was possible to predict prognosis because higher basal activity and serial rise were associated with poor prognosis. PMID- 16715613 TI - Mortality rates due to diabetes in a selected urban south Indian population--the Chennai Urban Population Study [CUPS--16]. AB - OBJECTIVE: The aim of this study was to determine the mortality rate in diabetic and non-diabetic subjects in urban south India. METHODS: The Chennai Urban Population Study is an ongoing epidemiological study in Chennai [formerly Madras, in south India]. All individuals > or = 20 years of age living in two residential colonies in Chennai were invited to participate in the study. Of the total 1399 eligible subjects, 1262 individuals responded [90.2%] at baseline, and of these, 1140 individuals [90.3%] could be followed annually from 1997 to 2003-04. Mortality rates and causes of death were the main outcome measures. RESULTS: The median follow up period was six years. The overall mortality rate was higher in diabetic compared to non-diabetic subjects [18.9 vs.5.3 per 1000 person-years, p=0.004]. Mortality due to cardiovascular [diabetic subjects, 52.9%; non-diabetic subjects, 24.2%,p=0.042] and renal causes[diabetic subjects, 23.5%; non-diabetic subjects, 6.1%,p=0.072] was higher among diabetic subjects whereas mortality due to gastrointestinal 112.1%], respiratory [9.1%], lifestyle related [6.1%] and unnatural causes [18.2%] were observed only among non-diabetic subjects. Hazards ratio [HR] for all cause mortality for diabetes was 3.6, [95% Confidence Interval [CI]: 2.02-6.53, p<0.001] and this remained significant even after adjusting for age [HR:1.9, 95% CI:1.04-3.45, p=0.038]. Light grade physical activity was associated with higher mortality rate [p=0.008], but the significance disappeared when adjusted for age. Smoking was also associated with increased mortality. CONCLUSIONS: In urban India, mortality rates are two fold higher in people with diabetes compared to nondiabetic subjects. Cardiovascular and renal diseases are the commonest causes of death among diabetic subjects. PMID- 16715615 TI - Phylogenetic tree construction of butyrylcholinesterase sequences in life forms. AB - Butyrylcholinesterase is an enzyme with few known physiological functions. It is related to acetylcholine that was shown to be expressed in a variety of life forms. We performed a search using the human butyrylcholinesterase gene (HGNC:983;MIM:177400), and found the sequence in a broad spectrum including plants, bacteria and animals. Therefore butyrylcholinesterase appears to have evolved early in evolution, and to have been conserved. PMID- 16715616 TI - Right atrial myxoma. PMID- 16715617 TI - Ruptured sinus of valsalva into right ventricle: a unique case with prolonged survival. PMID- 16715618 TI - Marcello Malpighi (1628-1694). Postal stamps released on Malpighi, Italy--1978 and Transkei--1985. PMID- 16715619 TI - Hypertension as a low-grade systemic inflammatory condition that has its origins in the perinatal period. AB - Genetics, oxidative stress: superoxide anion (O2*-) and hydrogen peroxide (H2O2), endothelial nitric oxide (eNO), lipid peroxides, anti-oxidants, endothelin, angiotensin converting enzyme (ACE) activity, angiotensinII, transforming growth factor-beta (TGF-beta), insulin, homocysteine, asymmetrical dimethyl arginine, proinflammatory cytokines: interleukin-6 (IL-6), tumor necrosis factor-a (TNF alpha), C-reactive protein (hs-CRP), and long-chain polyunsaturated fatty acids (LCPUFAs), and activity of NAD(P)H oxidase have a role in human essential hypertension. There is a close interaction between endogenous molecules: eNO, endothelin, cytokines, and nutrients: folic acid, L-arginine, tetrahydrobiopterin (H4B), vitamin B6, vitamin B12, vitamin C, and LCPUFAs. Statins mediate some, if not all, of their actions through LCPUFAs, whereas these fatty acids (especially omega-3 fatty acids) suppress cyclo-oxygenase activity and the synthesis of pro inflammatory cytokines, and activate parasympathetic nervous system, actions that reduce the risk of major vascular events. Some LCPUFAs form precursors to lipoxins and resolvins that have anti-inflammatory actions. Low-grade systemic inflammation seen in hypertension seems to have its origins in the perinatal period and availability of adequate amounts of LCPUFAs during the critical periods of brain growth prevents the development of hypertension. This indicates that preventive strategies aimed at decreasing the incidence of hypertension and its associated conditions such as atherosclerosis, type 2 diabetes, coronary heart disease (CHD), and cardiac failure in adulthood need to be instituted during the perinatal period if they are to be effective. PMID- 16715620 TI - Umbilical cord blood transplantation: newer trends. AB - During last ten years, over 4000 umbilical cord blood transplantations have been performed worldwide. The interest in this modality of transplantation has been growing as this provides easy access to an alternative source of stem cells for treating cancer and serious genetic disorders with otherwise fatal outcome or immense morbidity. Umbilical cord blood is a commonly discarded source of useful stem cells. The outcome of transplantation using cells from this source in children mirrors the results of unrelated donor transplantation and hence the procedure is widely accepted by paediatric transplant community. Results are, however, hampered in adults due to low cell dose. Newer techniques, such as pooled or sequential cord blood transplantation, may help to increase progenitor cell numbers and improve immune reconstitution. In near future, non haematopoietic uses will make this even more exiting area. In this write-up, we will review this treatment including cord blood banking issues and the ethical concerns. We will discuss both paediatric and adult transplantations including certain new indications. PMID- 16715621 TI - Acute myositis. AB - Two cases of acute myositis characterized by intense muscle pain, weakness, myoglobinuria in one and muscle biopsy showing inflammatory changes in both with good recovery are described. It is reported because of the rarity of this condition. PMID- 16715622 TI - Coexistence of Takayasu's arteritis with ulcerative colitis. AB - The association of ulcerative colitis with Takayasu's arteritis is rarely reported. The occurrence of the two together is possibly related to a common pathophysiology involving alteration in immune mechanisms. Takayasu's arteritis is more prevalent in Japan and South East Asia whereas Ulcerative Colitis is more in Western countries. The coexistence of these two diseases is uncommon and hence this report. PMID- 16715623 TI - Recurrent Kimura's disease: excellent response to cyclosporine. AB - Kimura's disease is a chronic inflammatory disorder involving the skin, subcutaneous tissues and lymph nodes, predominantly in the head and neck region. Though surgery, intralesional or systemic steroids or radiation therapy have been the mainstay of treatment recurrence is a common problem. On the basis of occasional case report of Kimura's disease responding to cyclosporine, we attempted oral cyclosporine in our patient with dramatic improvement. PMID- 16715624 TI - Klippel Trenaunay syndrome. AB - Klippel-Trenaunay syndrome (KTS) is an uncommon entity. This congenital malformation is characterized by the triad of soft tissue or bony hypertrophy, cutaneous vascular malformations, and atypical venous abnormalities. We report here a case of KTS and discuss the clinical features, investigations, and management of this enigmatic condition. PMID- 16715625 TI - Evaluation of typhoid assay for rapid diagnosis of typhoid fever. PMID- 16715626 TI - Spectrum of hemoglobinopathies in the state of Orissa: a ten years cohort study. PMID- 16715627 TI - Inverse correlation between HLA-DR antigen expression and CD4 positive lymphocytic populations in normal mucosa, tubulovillous adenoma, and invasive carcinoma of the colon. AB - BACKGROUND: HLA-A,B,C and HLA-D molecules present antigenic peptides to the antigen-specific receptor of autologous T lymphocytes. T-cell-mediated host versus-tumor response might therefore depend on the presence of these molecules on tumor cells, although the absence of HLA-A,B,C determinants on a cell has been shown to increase its susceptibility to lysis by natural killer cells. The prognostic role of tumor stage and grade is well- established in colorectal cancer. In this study we used immunohistochemistry to analyse the expression of HLA-DR on epithelial cells of normal colonic mucosa, tubulovillous adenoma, and invasive carcinoma, as well as the magnitude of the stromal T lymphocytes at the relevant sites. HLA-DR expression was correlated to histological grade and Dukes stage in the cases of invasive cancer. Yet, we investigated the association of HLA-DR plus DQ genes and adenoma or carcinoma by PCR. MATERIALS AND METHODS: 31 cases of normal colonic mucosa, 12 cases of tubulovillous adenoma, and 39 cases of invasive carcinoma were surveyed for the detection of HLA-DR monoclonal antigen, and the T helper (TH) marker (CD4) in the stroma (lamina propria) of the relevant cases. RESULTS: HLA-DR was expressed in 20 of 31 normal colonic mucosas (64.5%), 4 of 12 adenomas (33.3%), and in 10 of 39 invasive carcinomas (25.6%). A strong relation of HLA-DR expression and histological grade was found (p < 0.001), but no association with Dukes stage (p = 0.141). No significant correlation between HLA-DR plus DQ genes and adenoma or cancer of the colon was found. CD4 positive cells were found in 9 of 31 normal colonic mucosas (29%), 5 of 12 adenomas (42%), and in 26 of 39 invasive carcinomas (67%). CONCLUSIONS: The results showed an inverse correlation between the expression of HLA-DR and the number of CD4 positive cells as the lesion progressed to malignancy. HLA-DR was significantly associated with tumor grade but not with Dukes stage in colonic cancer hosts. HLA-DR and DQ genes do not contribute to a susceptibility to adenoma or carcinoma. PMID- 16715628 TI - [Diagnostic procedures in skin adnexal tumours]. AB - From histogenetic, morphologic and immunohistochemical point of view the authors try to make possible algorithms that can be employed in a routine diagnosis of adnexal skin tumours. They stress the importance of knowledge of clinical data necessary for orientation classification of tumour skin lesions after biopsies. The authors translated their obtained data into survey tables to be used as guidelines in a routine bioptic practice. PMID- 16715629 TI - [What is your diagnosis? Answer: paraganglioma of the filum terminale/cauda equina]. PMID- 16715630 TI - Argyrophilic grain disease: case report of the first two cases in the Czech Republic and review of the literature. AB - Argyrophilic grain disease (AgD) is a relatively newly described neurodegenerative disease with late-onset dementia. Morphologically it is characterized by the presence of abundant spindle-shaped argyrophilic grains (ArG) in neuronal processes and coiled bodies in oligodendrocytes. ArG consist of abnormally hyperphosphorylated form of tau protein. AgD is a substrate of at least 5% of all dementia cases with increasing incidence in the old age. Here we report the cases of a 91-year-old woman and an 83-year-old man clinically diagnosed with dementia. Neuropathological, histochemical and immunohistochemical examination of the brain tissue show the changes to be compatible with a definite diagnosis of AgD. This is the first description of two cases of AgD in the Czech Republic. PMID- 16715631 TI - [Mammary carcinoma with osteoclast-like giant cells--report of two cases]. AB - Authors describe two cases of a rare type of mammary carcinoma occurring in 47 year and 49-year-old women, characterised by osteoclast-like giant cells, hypervascular stroma, and signs of older and recent haemorrhages on the background of the conventional type of breast carcinoma. In both cases there was diagnosed moderately differentiated ductal invasive carcinoma with osteoclast like giant cells, which were present in tumorous stroma as well as scattered between tumorous cells. Immunohistochemical analysis (vimentin and CD68 positivity and negativity of cytokeratins, EMA and S-100 protein) confirmed a histiocytic origin of these giant cells as well as their reactive non-neoplastic nature. Clinical and morphological features of this unusual type of breast carcinoma indicate that it should be regarded as an unique clinical- histomorphological entity with probably better prognosis when compared to ductal invasive carcinoma not otherwise specified and it should not be included among metaplastic breast carcinomas. PMID- 16715632 TI - [The subacute angiohypertrophic myelomalacia Foix-Alajouanine--a rare disease?]. AB - The subacute angiohypertrophic myelomalacia Foix-Alajouanine was first described by French authors in 1926 and belongs to the wider category of CNS vascular malformations. Both individual casuistics and larger sets of patients with this disease can be found in literature scarcely. We are presenting three necroptic cases of subacute angiohypertrophic myelomalacia (Foix-Alajouanine) that were mistakenly diagnosed clinically, once as metastatic lesion of spinal canal, once as amyotrophic lateral sclerosis, and once as inflammatory lesion of spinal cord. The histologic finding is dominated by myelomalacia of varying extent with varicosely dilated, constricted or even occluded arterialized both extra- and intramedullary veins, sporadically coupled with secondary thrombi. Clinical diagnostics is difficult and requires spinal angiography. According to literature data, it is presumable that this disease is not as rare as it may seem at first sign. PMID- 16715633 TI - Dedifferentiated mixed stromal-smooth muscle tumor of the uterus. Report of a case. AB - So-called dedifferentiation in mesenchymal neoplasms of the uterus is very rare. Among conventional low-grade stromal tumors only three cases of dedifferentiation were reported, whereas in mixed stromal-smooth muscle tumors the dedifferentiation was yet not described. Here we present such a case of low-grade mixed stromal-smooth muscle tumor with dedifferentiation. The tumor occurred in 52-years-old postmenopausal patient. The high-grade component representing a dedifferentiation showed morphology of undifferentiated sarcoma with myxoid change. The low-grade component with morphology of mixed stromal-smooth muscle tumor was limited to a few peripheral areas of the lesion. Immunohistochemically, the low-grade component showed typical positivity for CD10, estrogen receptor, progesterone receptor, and focal reactivity for myoid markers, whereas the dedifferentiated component expressed only vimentin, CD10 and estrogen receptor. This case demonstrates that low-grade mixed stromal-smooth muscle tumor of the uterus can dedifferentiate like a pure stromal tumor. It shows that extensive sampling/histological search may be needed for recognition of a minor component in a dedifferentiated tumor. PMID- 16715634 TI - [Epithelioid hemangioma of the foot]. AB - Epithelioid hemangioma (angiolymphoid hyperplasia with eosinophilia, EH/ALHE) is a rare benign angioproliferative lesion which typically occurs in the region of the head and neck. In the literature, occurence on the extremity is only rarely described. A case of multiple occurence of EH/ALHE in the skin of the toes and metatarsal bone with osteolysis is reported. Occurence on the extremity, superficial and deep affection and some "atypical" microscopic features may cause diagnostic dilemma. The key diagnostic features of EH/ALHE are vascular channels lined with epithelioid endothelial cells, surrounding layer of myopericytes, absence of atypia and mitotic activity and characteristic inflammation. Immunohistochemistry may be helpful in settling the diagnosis. PMID- 16715635 TI - [Tumoriform endometriosis of the urinary bladder in advanced pregnancy]. AB - A case of tumoriform endometriosis of urinary bladder immitating a tumor of the bladder during the first pregnancy of a 25-year-old patient was successfully treated by partial cystectomy allowing continuation of the pregnancy, a normal term delivery and after 5 months a second pregnancy with term delivery. PMID- 16715636 TI - [Observations of wounds]. PMID- 16715637 TI - [Update on dressings]. PMID- 16715638 TI - [Update on the value of hyaluronic acid for active wound healing]. PMID- 16715639 TI - Evaluating a school skills programme for Australian indigenous children: A pilot study. AB - The purpose of this study was to evaluate the effectiveness of an occupational therapy skill development programme in improving handwriting, scissor use, visual motor co-ordination and classroom behaviour, in a group of grade-one Australian urban Indigenous students. The sample (N = 13) was randomly assigned to an experimental group or comparison group. Both groups were exposed to the intervention. The experimental group received a six week school skills programme in addition to regular schooling, while the comparison group received regular schooling only. Following this, the comparison group received the same six week programme. Participants underwent pre- and post-testing using Beery-Buktenica's Developmental Test of Visual Motor Integration, Conner's Abbreviated Symptom Questionnaire, and two non-standardized handwriting and scissor skill measures. Results indicated that participants significantly improved in aspects of handwriting, scissor skills, and behaviour, but not visual motor co-ordination, following participation in the programme. The experimental group demonstrated greater improvements only in handwriting ability scores (p = 0.037), compared to a comparison group. It was concluded that a school-based occupational therapy programme was effective in improving handwriting, in a group of grade-one Australian Indigenous children. PMID- 16715640 TI - A pilot study of the activity patterns of five elderly persons after a housing adaptation. AB - The importance of an accessible home environment for occupational performance has been emphasized in occupational therapy, but knowledge about how accessibility can affect a person's activity patterns is limited. The purpose of this study was to describe the activity patterns of five elderly persons aged between 70 and 84 years and their views about accessibility and usability of their homes before and after a housing adaptation. Two different questionnaires, 'Accessibility in My Home' and the 'Occupational Questionnaire' were used for data collection. Adaptations in the home included: removing thresholds, installing new taps in the bathroom and kitchen and broadening doorways. After the adaptations the five participants reported that they increased their outside activities, reduced naps during the day and slept better at night. The results showed that the participants performed more and new activities when their home environment had become accessible. Activities performed were also perceived as more important after the housing adaptation. There is a need to replicate the study with a larger sample. Further research is needed on the impact of home adaptations on the activity patterns of elderly people. PMID- 16715641 TI - Frequency of knowledge of results and motor learning in persons with developmental delay. AB - The purpose of this study was to investigate the effect of high versus low frequency knowledge of results (KR) in a group of 16 individuals with developmental delay and in gender and age-matched average individuals learning a motor skill on a laptop computer. Participants were randomly assigned to either a 100% KR or a 50% KR group. KR was provided during the acquisition phase according to group assignment as participants learned the motor skill, whereas no KR was provided during the retention phase. Results indicated both populations who received 50% KR in the acquisition phase demonstrated better performance in the retention phase than those who received 100% KR. The results of this study suggest that, as has been found in the average population, feedback that is too frequent can interfere with learning and retention of tasks for individuals with developmental disabilities (DD). Limitations involved the small sample size along with the task potentially being artificial in nature. Future research is needed to study further the effects of frequency of KR on skill acquisition, particularly in instrumental activities of daily living in this population. PMID- 16715642 TI - Development of critical thinking in occupational therapy students. AB - Do students who use the Guided Reciprocal Peer Questioning (GRPQ) method increase their ability to think critically? When compared to students in a traditional senior seminar course, the results of this study indicated no significant difference between the groups regarding changes in scores on the California Critical Thinking Skills Test. However, the students in the experimental group asked more questions labelled as critical thinking than the seminar control group. These findings suggest the use of the GRPQ may improve students' skills in asking questions that include application, analysis, and synthesis. Future research regarding the role of questions in stimulating critical analysis and the role of context in the learning environment is warranted. PMID- 16715643 TI - Long-acting beta-agonists role in asthma management. AB - The LABAs have played an important role in the management of asthma over the past decade. They are of clear benefit in reducing asthma-related symptoms and improving lung function when used in combination with an anti-inflammatory agent. Studies have shown, however, that their use has been associated with various negative outcomes, which has led to a restricted indication for salmeterol xinafoate (Serevent Diskus) and fluticasone propionate and salmeterol xinafoate (Advair Diskus), along with medication guides that will be given to patients with every new and refill prescription. Convincing data now exist that show an association of salmeterol with an increase in asthma-related deaths and life threatening experiences, while formoterol is associated with more frequent serious asthma exacerbations. Nonetheless, LABAs remain an important component of asthma therapy. Further clarification about their role may occur with the release of additional analyses from SMART, as well as updated guidelines from the NAEPP Expert Panel, both expected later in 2006. In the meantime, LABAs and LABA containing products are to be used only for patients not adequately controlled on other asthma-controller medications (eg, low- to medium-dose inhaled corticosteroids) or whose disease severity clearly warrants initiation of treatment with 2 maintenance therapies. The NHLBI/NAEPP guidelines recommend inhaled corticosteroids as the first step in controller therapy, with LABAs as an option if low- to medium-dose inhaled corticosteroids do not adequately control the patient's asthma". PMID- 16715644 TI - [Expert witness and expert testimony in New Slovak Criminal Procedure Code]. AB - As of 1st January 2006, new criminal law consisting of the Act No. 300/2004 Coll. (Penal Code) and the Act No. 301/2005 Coll. (Criminal Procedure Code) became effective in the Slovak Republic. These two acts replaced Criminal Law originating from 1961. The author analyses impact and changes introduced with new Criminal Procedure Code in the regulation of expert witness in general and expert witness testimony. PMID- 16715645 TI - [The comparison of the enzymatic and the acid hydrolyse at the toxicological screening of the small amount of urine]. AB - The article deals with the toxicological screening directed at the identification and confirmation of the acid, neutral and basic medicaments and drugs in the urine. The efficiency of the hydrolysation using beta-glukuronidase (enzymatic hydrolyses) and using 0.1M hydrochloric acid (acid hydrolyses) from 2 ml of the urine were compared. The solid phase extraction (SPE) and the GC-MS identification were used. The efficiency of the extraction of paracetamol, buprenorphine and morphine is better by means of the enzymatic hydrolyses. Vice versa the acid hydrolyses appears to be more applicable for benzodiazepines. The acid hydrolyses is faster and cheaper and the extracts contain less of the ballast materials. PMID- 16715646 TI - [PCR-multiplex system for analysis of Y-chromosomal microsatellite polymorphisms DYS449, DYS456, DYS458, and DYS464]. AB - DNA profiling--inclusive sex determination-- with microsatellite markers is currently a commonly used genetic method of studying humans. An efficient technique of producing the genetic data is amplification of multiple microsatellites in a single PCR reaction. Here we introduce a novel PCR-multiplex system for analysis of four polymorphic Y-STRs. Specifically, these are DYS449, DYS456, DYS458, and DYS464. These loci were chosen because of their reported high diversity in Euroamerican population (10), as well as their absence in the commercial analytical kits at the time of beginning of this study. Our objective was to design this PCR-multiplex for use of fragmentation analysis by electrophoresing samples on a capillary semi-automated genetic analyzer applying only one fluorescent dye. The PCR system we propose, may be notably used in fields such as forensic and human population genetics. PMID- 16715647 TI - Basic characteristics of chronic hypotension cases: a longitudinal follow-up study from 1958 through 1999. AB - Few epidemiological studies have been conducted on the prevalence, blood pressure trends by age, and basic characteristics of chronic hypotension. Among 13,370 subjects (5,094 men and 8,276 women) examined from 1958 to 1999 in Japan, 92 were identified as chronic hypotensives (11 men and 81 women) whose systolic blood pressure (SBP, mmHg) was lower than 100 mmHg for 8 or more years. Trends of SBP and body mass index (BMI, kg/m2) were determined over a 16-year period in this group of chronic hypotensives and 276 age- and sex-matched controls (33 men and 243 women); during this 16-year period, the chronic hypotensives and controls ranged in age from 19 to 75 years. BMI, heart rate (HR, bpm), body temperature (degrees C), Hb (g/dl), and creatinine (Cre, mg/dl) were compared between the chronic hypotensives and the controls at the last examination. The chronic hypotensives had a female preponderance (0.2% in men and 1.0% in women, p=0.001), and their SBP and BMI increased little with increasing age. BMI (20.2+/-3.4), BMI adjusted SBP (101+/-19), HR (63+/-10), body temperature (36.7+/-0.3), Hb (12.5+/ 1.1), and Cre (0.81+/-0.13) were lower in the chronic hypotensives than those (22.9+/-3.4, 126+/-20, 68+/-10, 36.8+/-0.3, 12.9+/-1.1, and 0.86+/-0.12, respectively) in controls (p<0.05). The present results clearly show that chronic hypotensives have a female preponderance and low BMI, HR, body temperature, Hb, and Cre in addition to low SBP that does not increase with age. PMID- 16715648 TI - Pathophysiological roles of the adrenal renin-angiotensin system in patients with primary aldosteronism. AB - The mechanism of overproduction of aldosterone in primary aldosteronism is unclear. The intraadrenal renin-angiotensin system (RAS) has been suggested to possess the functional role of the synthesizing aldosterone and regulating blood pressure. In order to clarify the pathophysiological roles of adrenal RAS in aldosterone-producing adenoma (APA), we studied the expressions of the messenger RNAs (mRNAs) of renin, angiotensinogen, type 1 (AT1R) and type 2 angiotensin II receptor (AT2R), CYP11B1 (11 beta-hydroxylase gene) and CYP11B2 (aldosterone synthase gene) in 8 patients with angiotensin II-responsive (ATII-R) APA and compared them with the expressions of the same mRNAs in 8 patients with angiotensin II-unresponsive (ATII-U) APA. Quantification of the mRNA of each gene was done using a real-time polymerase chain reaction with specific primers. There were no significant differences between ATII-R APA and ATII-U APA in the mRNA levels of renin, angiotensinogen, AT1 R, CYP11B1 and CYP11B2. The amount of AT2R mRNA was significantly higher in the patients with ATII-R APA than in those with ATII-U APA (p<0.05). These results may suggest that AT2R partially contributes to the overproduction of aldosterone in ATII-R APA. PMID- 16715649 TI - Impact of hypertension and hypertension-related vascular lesions in IgA nephropathy. AB - It remains poorly understood whether vascular pathology plays an important role in the progression of renal parenchymal disease in humans. Moreover, in the case of hypertensive patients with mild proteinuria, nephrologists tend to make a diagnosis of benign nephrosclerosis without renal biopsy. Among 172 patients who were treated at our hospital for biopsy-proven IgA nephropathy, we performed quantitative histopathological analysis in 38 patients with mild proteinuria of less than 0.5 g/day. We related these histopathological parameters with clinical data at biopsy and also with follow-up data. The percentage of glomeruli showing global sclerosis exceeded 10% of total glomeruli in 15 of the patients (39.5%) and exceeded 20% in 9 (23.7%). Arteriosclerosis and tubulointerstitial changes significantly correlated with glomerular sclerosis, but mesangial cell proliferation did not. Among the 38 patients, the 12 with hypertension showed more severe glomerular sclerosis, tubulointerstitial changes and arteriosclerosis compared with the 26 without hypertension, but the mesangial cell proliferation was identical between the two groups. Stepwise multiple regression analysis revealed that hypertension and urinary protein excretion (UPE) were independent risk factors for arteriosclerosis. The follow-up data of a mean period of 27.6 months showed that 9 of the 38 patients (23.7%) had an increase in UPE. Hypertension, arteriosclerosis, age, and UPE at biopsy were selected as the important risk factors for an increase in UPE in the follow-up. Our results provide not only clinical but histopathological evidence that hypertension affects the prognosis of mild proteinuric nephropathy through vascular lesions. PMID- 16715650 TI - Ankle brachial index as a marker of atherosclerosis in Chinese patients with high cardiovascular risk. AB - To obtain reliable data on the epidemiology, co-morbidities and risk factor profile of peripheral arterial disease (PAD), we evaluated the clinical significance of the ankle brachial index (ABI) as an indicator of PAD in Chinese patients at high cardiovascular (CV) risk. ABI was measured in 5,646 Chinese patients at high CV risk, and PAD was defined as an ABI<0.9 in either leg. Multivariable logistic regression analyses were performed to identify factors associated with PAD. A total of 5,263 patients were analyzed, 52.9% male, mean age 67.3 years, mean body mass index (BMI) 24.2 kg/m2, mean systolic/diastolic blood pressure (SBP/DBP) 139/80.7 mmHg. The prevalence of PAD in the total group of patients was 25.4%, and the prevalence was higher in females than in males (27.1% vs. 23.9%; odds ratio [OR]: 1.64). Patients with PAD were older than those without PAD (72.3+/-9.9 years vs. 65.6+/-11.7 years; OR: 1.06), and more frequently had diabetes (43.3% vs. 31.3%; OR: 2.02), coronary heart disease (CHD) (27.0% vs. 18.8%; OR: 1.67), stroke (44.4% vs. 28.3%; OR: 1.78), lipid disorders (57.2% vs. 50.7%; OR: 1.3) and a smoking habit (42.7% vs. 38.6%; OR: 1.52). The ORs for the PAD group compared with the non-PAD group demonstrated that these conditions were inversely related to ABI. Statin, angiotensin-converting enzyme inhibitors and antiplatelet agents were only used in 40.5%, 53.6% and 69.1% of PAD patients, respectively. The data demonstrated the high prevalence and low treatment of PAD in Chinese patients at high CV risk. A lower ABI was associated with generalized atherosclerosis. Based on these findings, ABI should be a routine measurement in high risk patients. Aggressive medication was required in these patients. PMID- 16715651 TI - Impact of metabolic syndrome on brachial-ankle pulse wave velocity in Japanese. AB - The aim of this study was to determine the effect of metabolic syndrome on brachial-ankle pulse wave velocity (baPWV) by using the new guidelines for diagnosis of this syndrome in Japan. We examined 525 men and women without a history of cardiovascular disease or cancer, and an ankle-brachial index < 0.9. The baPWV was measured using a device (Form PWV/ABI) that simultaneously monitored bilateral brachial and ankle pressure wave forms. Metabolic syndrome was defined as a waist circumference > or = 85 (90) cm in men (women) and two or more of the following risk factors: hypertension, dyslipidemia, and glucose intolerance diagnosed by a 75 g oral glucose tolerance test. The baPWV showed a significant linear relationship with waist circumference, waist-to-hip ratio, body fat, systolic and diastolic blood pressure, triglycerides, fasting glucose, 2-h-postload glucose, fasting insulin, and glycosylated hemoglobin-A1c, after adjusting for sex and age. These factors were also strongly related to fasting insulin levels. When subjects were classified into six groups based on waist circumference and the number of risk factors for metabolic syndrome (0, 1, and > or =2), we found that more risk factors clearly increased the odds ratios for an elevated baPWV in those subjects in the highest quartile of the baPWV distribution in multivariate logistic models. An increase in odds ratio was observed despite a normal waist circumference and may well have been due to increased fasting insulin and blood pressure levels. An increase in the number of risk factors for metabolic syndrome was highly correlated with an increased baPWV, probably due to insulin resistance. PMID- 16715652 TI - CAMP-response element-binding protein mediates tumor necrosis factor-alpha induced vascular cell adhesion molecule-1 expression in endothelial cells. AB - Hypertension causes endothelial dysfunction, which plays an important role in atherogenesis. The vascular cell adhesion molecule-1 (VCAM-1) contributes to atherosclerotic lesion formation by recruiting leukocytes from blood into tissues. Tumor necrosis factor-alpha (TNFalpha) induces endothelial dysfunction and VCAM-1 expression in endothelial cells (ECs). We examined whether the cAMP response element binding protein (CREB), a transcription factor that mediates cytokine expression and vascular remodeling, is involved in TNFalpha-induced VCAM 1 expression. TNFalpha induced phosphorylation of CREB with a peak at 15 min of stimulation in a dose-dependent manner in bovine aortic ECs. Pharmacological inhibition of p38 mitogen-activated protein kinase (p38-MAPK) inhibited TNFalpha induced CREB phosphorylation. Adenovirus-mediated overexpression of a dominant negative form of CREB suppressed TNFalpha-induced VCAM-1 and c-fos expression. Although activating protein 1 DNA binding activity was attenuated by overexpression of dominant negative CREB, nuclear factor-kappaB activity was not affected. Our results suggest that the p38-MAPK/CREB pathway plays a critical role in TNFalpha-induced VCAM-1 expression in vascular endothelial cells. The p38MAPK/CREB pathway may be a novel therapeutic target for the treatment of atherosclerosis. PMID- 16715653 TI - Amlodipine-induced reduction of oxidative stress in the brain is associated with sympatho-inhibitory effects in stroke-prone spontaneously hypertensive rats. AB - Amlodipine is a dihydropyridine calcium channel blocker that is widely used for the treatment of hypertensive patients and has an antioxidant effect on vessels in vitro. The aim of the present study was to examine whether treatment with amlodipine reduced oxidative stress in the brains of stroke-prone spontaneously hypertensive rats (SHRSP). The animals received amlodipine, nicardipine or hydralazine for 30 days in their drinking water. Levels of thiobarbituric acid reactive substances (TBARS) in the brain (cortex, cerebellum, hypothalamus, and brainstem) were measured before and after each treatment. Systolic blood pressure decreased to similar levels in the amlodipine-, nicardipine-, and hydralazine treated groups. Urinary norepinephrine excretion was significantly reduced in SHRSP after treatment with amlodipine, but not with nicardipine or hydralazine. Levels of TBARS in the cortex, cerebellum, hypothalamus, and brainstem were significantly higher in SHRSP than in Wistar-Kyoto rats (WKY), and were reduced in amlodipine-treated, but not in nicardipine- or hydralazine-treated, SHRSP. Electron spin resonance spectroscopy revealed increased levels of reactive oxygen species in the brains of SHRSP, which were reduced by treatment with amlodipine. Intracisternal infusion of amlodipine also reduced systolic blood pressure, urinary norepinephrine excretion, and the levels of TBARS in the brain. These results suggested that oxidative stress in the brain was enhanced in SHRSP compared with WKY rats. In addition, antihypertensive treatment with amlodipine reduced oxidative stress in all areas of the brain examined and decreased blood pressure without a reflex increase in sympathetic nerve activity in SHRSP. PMID- 16715654 TI - Chymase inhibition prevents cardiac fibrosis and dysfunction after myocardial infarction in rats. AB - Human chymase activates not only angiotensin II but also transforming growth factor-beta, a major stimulator of myocardial fibrosis, while rat chymase activates transforming growth factor-beta, but not angiotensin II. To clarify the role of chymase-dependent transforming growth factor-beta activation, we evaluated whether chymase inhibition prevents cardiac fibrosis and cardiac dysfunction after myocardial infarction in rats. Myocardial infarction was induced by ligation of the left anterior descending coronary artery. One day after the ligation, rats were randomized into 2 groups: 1) a chymase-treated group that received 10 mg/kg per day of the chymase inhibitor NK3201 orally for 4 weeks; and 2) a vehicle group of non-treated rats with myocardial infarction. We also included a control group who underwent sham-operation and no treatment. Four weeks after ligation, echocardiography revealed that chymase inhibitor treatment reduced the akinetic area and increased fractional area change but did not significantly change left ventricular end-diastolic area. Chymase inhibition significantly reduced left ventricular end-diastolic pressure, increased the maximal end-systolic pressure-volume relationship and decreased the time constant of left ventricular relaxation. Chymase activity in the non-infarcted myocardium was significantly increased in the vehicle group, but it was significantly reduced by chymase inhibitor treatment. The fibrotic area in the cardiac tissues and the mRNA levels of collagen I and collagen III were also significantly lower in the chymase inhibitor-treated group than in the vehicle group. Therefore, the pathway forming chymase-dependent transforming growth factor-beta may play an important role in myocardial fibrosis and cardiac dysfunction rather than left ventricular dilatation after myocardial infarction. PMID- 16715655 TI - Perineural spread in head and neck malignancies. AB - Perineural tumor spread (PNS) of head and neck malignancies is a well-known form of metastatic disease in which a lesion can migrate away from the primary site along the endoneurium or perineurium. This pattern of spread may create a poor prognosis and require aggressive treatment when curable. Although representative histologies are squamous cell carcinoma and adenoid cystic carcinoma, other malignancies such as malignant lymphoma and sarcoma also can show such a specific pattern of extent. PNS can be insidious, often delaying diagnosis. Knowledge of anatomy of the nerves is crucial in the imaging diagnosis of PNS, to detect early curable disease. The facial nerve and the maxillary and mandibular divisions of the trigeminal nerve are most commonly affected. General clinical issues and the diagnostic imaging of PNS along these nerves are discussed in the current article. PMID- 16715656 TI - Results of primary central nervous system lymphoma treated by radiation and chemotherapy: retrospective analysis of twelve institutions in the Tokai District of Japan, 1995-1999. AB - PURPOSE: We analyzed the therapeutic results and prognostic factors of 46 primary central nervous system lymphoma (PCNSL) patients who were treated at twelve institutions in the Tokai district of Japan between 1995 and 1999. We compared the results with those of a Japanese nationwide survey performed in the past. MATERIALS AND METHODS: We sent each institution a questionnaire about the state of patients' disease, pathological type, method and doses of radiotherapy, regimen and intensity of chemotherapy, and patients' prognoses. The range of patients' ages was 33 to 93 years (median, 61 years). Thirty-one were men and 15 were women. The most prevalent histology was diffuse large B cell type (33 patients). We used the Kaplan-Meier method to calculate the survival rate and Cox's proportional hazards model to analyze the prognostic factors. RESULTS: The five-year cumulative survival rate was 25%, and the median survival time was 22.7 months. The five-year disease-free survival rate was 23%. In monovariate analysis, patients who were both younger than 60 years old and had a WHO performance status (PS) score equal to or less than 2 showed a better survival rate. Furthermore, the patients receiving systemic chemotherapy showed a significantly better local control rate. In addition, patients who received systemic chemotherapy achieved a higher complete remission rate than those not receiving it. However, no factors that significantly influenced survival rate were identified in multivariate analysis. CONCLUSION: We demonstrated that the therapeutic outcome of PCNSL patients has recently improved. In particular, patients with good PS showed better local control than those with poor PS. However, we could not identify any significant prognostic factors in PCNSL patients. PMID- 16715657 TI - Treatment planning and dosimetry of a multi-axis dynamic arc technique for prostate cancer: A comparison with IMRT. AB - PURPOSE: Intensity-modulated radiation therapy (IMRT) allows greater dose conformity to the tumor target. However, IMRT, especially static delivery, usually requires more time to deliver a dose fraction than conventional external beam radiotherapy (EBRT). The authors have been using a "two-axis dynamic arc therapy" (2A-DAT) technique for prostate cancer treatment to make a concave dose distribution to spare the rectum and bladder while working with limited time and human resources. The objectives of this study were to (1) clinically implement the 2A-DAT technique, (2) evaluate the dosimetry in comparison with IMRT, and (3) analyze the initial treatment outcome. MATERIALS AND METHODS: The 2A-DAT consists of two dynamic arc therapies (DATs) with half rotation around two isocenters each in two separate symmetrical rhombi. Treatment planning is forward and on a trial and-error basis. Thirty-four patients received 2A-DAT with a median prescribed dose of 70 Gy. RESULTS: Although inferior in dose uniformity, the 2A-DAT provided equivalent sparing of normal structures to IMRT. Daily fraction delivery time for the 34 patients ranged from 6.4 to 9.6 minutes, with an average of 7.4 minutes. Five-year survival and five-year prostate specific autigen (PSA) failure-free survival were 89.3% and 79.5%, respectively. Three patients developed grade 2 proctitis. CONCLUSION: This technique is a possible alternative to IMRT in EBRT of prostate cancer. PMID- 16715658 TI - Comparison of the MR findings on indirect MR arthrography in patients with rotator cuff tears with and without symptoms. AB - PURPOSE: To compare indirect magnetic resonance (MR) arthrography findings in patients with rotator cuff tears with and without symptoms. MATERIALS AND METHODS: Indirect MR arthrography results of 15 asymptomatic and 23 symptomatic rotator cuff tears were retrospectively reviewed, comparing the type and size of tears and amount of subacrominal-subdeltoid bursal fluid. RESULTS: There were 14 partial-thickness tears and one full-thickness tear in the asymptomatic group, with 4 small and 11 medium tears. In the symptomatic group, there were 16 partial thickness and 7 full-thickness tears, with 14 small, 5 medium, and 4 large tears. In the asymptomatic group, the amount of subacrominal-subdeltoid bursal fluid was grade 0, 6 patients; grade 1, 6 patients; grade 2, 2 patients; and grade 3, 1 patient. In the symptomatic group, the amount of subacrominal-subdeltoid bursal fluid was grade 0, 4 patients; grade 1, 3 patients; grade 2, 8 patients; and grade 3, 8 patients. We found no statistically significant difference between symptomatic and asymptomatic tears in the kind and size of tears, and in the amount of subacrominal-subdeltoid bursal fluid. CONCLUSION: We found no difference in the MR findings between symptomatic and asymptomatic rotator cuff tears. PMID- 16715659 TI - Novel display technique for reference images for visibility of temporal change on radiographs--color digital summation radiography. AB - PURPOSE: The present study investigated color digital summation radiography (CDSR) as a novel display technique for reference images for the visibility of temporal change on radiographs. In CDSR, only the parts with temporal differences are displayed in color. Moreover, all other parts are displayed in gray scale. MATERIALS AND METHODS: CDSR was defined as "radiographs combined by the additive color mixture method". The visibility of simulated nodules located in the lung fields and mediastinum was evaluated by 12 radiologists (mean experience, 9.8 years; range, 1-26 years) for 24 conventional radiographs and CDSR for 6 color patterns. A five-point rating system (5, very good; 4, good; 3, adequate; 2, poor; 1, very poor) was used. RESULTS: The mean scores (average +/- standard deviation) for the visibility of simulated nodules were as follows: magenta, 3.88 +/- 0.90; blue, 3.08 +/- 0.72; green, 3.04 +/- 0.86; red, 3.00 +/- 0.98; cyan, 2.71 +/- 0.86; and yellow, 2.50 +/- 0.72. Compared to conventional radiography in gray scale, at 1.21 +/- 0.41, all six color patterns for CDSR displayed significantly improved scores (p<0.001). CONCLUSION: CDSR might represent a useful technique for reference images from chest digital radiography. PMID- 16715660 TI - Dynamic helical CT mammography of breast cancer. AB - PURPOSE: The purpose of this study was to determine whether dynamic helical computed tomography (CT)-mammography could assist in selecting the most appropriate surgical method in women with breast cancer. MATERIALS AND METHODS: Preoperative contrast-enhanced helical CT scanning of the breast was performed on 133 female patients with suspicion of breast cancer at the same time as clinical, mammographic, and/or ultrasonographic examinations. The patients were scanned in the prone position with a specially designed CT-compatible device. A helical scan was made with rapid intravenous bolus injection (3 ml/s) of 100 ml of iodine contrast material. Three-dimensional maximum intensity projection (MIP) images were reconstructed, and CT findings were correlated with surgical and histopathological findings. RESULTS: Histopathological analysis revealed 84 malignant lesions and seven benign lesions. The sensitivity, specificity, and accuracy levels of the CT scanning were 94.6%, 58.6%, and 78.9%. Helical scanning alone revealed additional contralateral carcinomas in three of four patients and additional ipsilateral carcinomas in three of five patients. However, the technique gave false-positive readings in 24 patients. The preoperative CT mammogram altered the surgical method in six patients. CONCLUSION: Dynamic helical CT-mammography in the prone position may be one of the choices of adjunct imaging in patients with suspected breast cancer scheduled for surgery. PMID- 16715661 TI - Retrospective review: usefulness of a number of imaging modalities including CT, MRI, technetium-99m pertechnetate scintigraphy, gallium-67 scintigraphy and F-18 FDG PET in the differentiation of benign from malignant parotid masses. AB - OBJECTIVE: The aim of this study is to evaluate an imaging approach using computed tomography (CT), magnetic resonance imaging (MRI), technetium-99m pertechnetate scintigraphy (99mTc pertechnetate scintigraphy), gallium-67 scintigraphy (67Ga scintigraphy) and fluorine-18 fluorodeoxyglucose positron emission tomography (18F-FDG PET). MATERIAL AND METHODS: We reviewed retrospectively 59 patients with parotid masses. CT, MRI, 67Ga scintigraphy, 99mTc pertechnetate scintigraphy, and 18F-FDG PET were performed. RESULTS: All of the benign tumors had smooth margins on CT and MRI. Patients with inflammatory lesions and malignant lesions showed well-defined margins or ill-defined margins. All Warthin's tumors showed high technetium accumulation. Many of malignant tumors showed high FDG accumulation. Both pleomorphic adenomas and Warthin's tumors showed high accumulation in some cases on 18F-FDG PET. Among 15 patients with pleomorphic adenoma, 14 patients showed marked hyperintensity relative to CSF on T2-weighted images and partial enhancement on contrast-enhanced T1 weighted images. Combination of several imaging modarity offered usefulness of differential diagnosis for parotid masses. CONCLUSION: An efficient combination of imaging methods may be helpful for achieve the correct diagnosis. PMID- 16715662 TI - Intracavitary brachytherapy for carcinoma of the uterine cervix--comparison of HDR (Ir-192) and MDR (Cs-137). AB - PURPOSE: To compare the results of high dose rate (HDR) (Ir-192) and medium dose rate (MDR) (Cs-137) intracavitary brachytherapy (ICRT) for carcinoma of the uterine cervix. MATERIALS AND METHODS: Between May 1991 and March 2001, a total of 206 patients with Stage I-IVA previously untreated cervical cancer were treated with ICRT combined with external beam radiotherapy (EBRT). HDR was administered to a total of 135 patients: 22 patients in Stage I, 49 in Stage II, 56 in Stage III, and eight in Stage IVA. MDR was administered to a total of 71 patients: six patients in Stage I, 27 in Stage II, 33 in Stage III, and five in Stage IVA. The MDR at point A was 30 Gy/hour for HDR and 1.7 Gy/hour for MDR treatment, and the corresponding median follow-up periods for survivors were 55 and 68 months. RESULTS: For the HDR group, 5-year cause-specific survival rates were 90%, 78%, 53% and 33% for Stages I, II, III, and IVA, respectively. For the MDR group, the corresponding rates were 100%, 76%, 51%, and 40%. In the HDR group, 19 patients (14%) developed Grade 2 or higher late complications, and, in the MDR group, four patients (6%) did. CONCLUSIONS: There was no statistically significant difference in cause-specific survivals between the results of HDR and MDR brachytherapy for cervical cancer. The incidence of late complications tended to be higher for the HDR group than for the MDR group, but did not show a statistically significant difference (p=0.07). PMID- 16715663 TI - High-dose-rate brachytherapy combined with long-term hormonal therapy for high risk prostate cancer: results of a retrospective analysis. AB - PURPOSE: High-dose-rate (HDR) brachytherapy combined with hormonal therapy (HTx), without the addition of external beam radiation therapy (EBRT) for high-risk prostate cancer was evaluated retrospectively. MATERIALS AND METHODS: Between May 1995 and April 2002, 35 patients with prostate cancer [Stage > or = T2b (UICC 1997) or tumor grading=3 or prostate-specific antigen (PSA) level > or = 20 ng/mL] were treated with HDR brachytherapy combined with HTx. Most patients (74%) had two or more of these factors. All patients received Iridium-192 HDR brachytherapy with a total dose of 54 Gy/9 fractions/5 days (48 Gy/8 fractions/5 days for the first 6 cases) in one implant session. The median neoadjuvant HTx [luteinizing hormone-releasing hormone (LH-RH) agonist and antiandrogen] period was 7 months. The median adjuvant HTx (ATH) (LH-RH agonist) period was 40 months, and median follow-up was 57 months (range, 23-117 months). RESULTS: The 5-year actuarial biochemical control, local control, and disease-free rates were 62%, 96%, and 76% respectively. No patients experienced local and/or regional relapse without distant progression. The 5-year actuarial cause-specific survival and overall survival rates were 89% and 87%, respectively. The acute and late toxicity were moderate and well tolerated. CONCLUSION: HDR brachytherapy plus long-term HTx is at least as effective as conventional EBRT plus long-term HTx. PMID- 16715664 TI - Treatment results of preoperative concurrent chemoradiotherapy followed by surgery for stage III or IV esophageal squamous cell carcinoma. AB - PURPOSE: To evaluate the treatment outcome of preoperative neoadjuvant radiotherapy combined with chemotherapy (CTx) for 17 esophageal cancer patients. MATERIALS AND METHODS: Between 1992 and 2004, patients with locally advanced esophageal cancer (stage III or IV) before curative-intent surgery received radiotherapy (RT) combined with CTx (Cisplatin: 75 mg/m2, bolus infusion, and 5 fluorouracil (5-FU): 1,000 mg/m2/24 h, continuous infusion for 4 days) at a median total dose of 30 Gy (n=17). RESULTS: The median survival period was 13.8 months. The overall survival rates at 1, 2, and 3 years were 75%, 40%, and 20%, respectively. According to univariate analysis, no factor of worse prognosis was found. Pathological markedly (Grade 3) or moderately (Grade 2) effects were observed in 4 patients (24%) and 8 patients (47%), respectively. CONCLUSION: These results indicate that, although this regimen was effective in terms of pathological effect, it is unclear whether it made a contribution to the improvement of survival PMID- 16715665 TI - Evaluation of coronary artery bypass grafts using multidetector-row CT with Japanese patients. AB - PURPOSE: To evaluate the patency of coronary artery bypass grafting (CABG) with multidetector-row computed tomography (MDCT) in Japanese patients, who have narrower coronary arteries than Caucasians. MATERIALS AND METHODS: Nineteen patients (12 men and 7 women, mean age: 63 +/- 12 years) with 33 coronary bypass grafts were examined with three-dimensional volume rendering (3D-VR) and curved multiplanar reconstruction (MPR) of MDCT. All grafts were compared with those obtained with selective graft angiography. RESULTS: For the 19 patients with 33 grafts, the overall sensitivity and specificity, compared with those for selective bypass angiography, were 96.7% and 100%, respectively. CONCLUSION: The patency of CABG in Japanese patients can be effectively evaluated with 3D-VR and curved MPR of MDCT. PMID- 16715666 TI - Umbilical metastasis from breast cancer related with tumor marker elevation. AB - We describe a 53-year-old woman with tumor marker abnormality caused by an umbilical metastasis from breast cancer. She had undergone breast conservation therapy (BCT) for breast cancer (T2N1M0) 9 years previously. Umbilical metastasis was detected 9 months after tumor marker elevation was first noted. After resection of the umbilical metastasis, tumor marker level decreased immediately and normalized. The patient is alive without other metastasis. PMID- 16715667 TI - A new method using J-type long sheath for implantation of indwelling catheters for trans-femoral hepatic arterial infusion. AB - PURPOSE: For implantation of a catheter-port system for hepatic arterial infusion chemotherapy, catheterization via the femoral artery is occasionally difficult. The purpose of this study was to determine the usefulness of a J-type long sheath, which facilitates difficult trans-femoral catheterization. MATERIALS AND METHODS: We applied this technique to 15 patients for difficult trans-femoral catheterization for catheter-port system placement. RESULTS: We successfully implanted a catheter-port system in all patients using this technique without significant complications. CONCLUSION: We emphasize that our new technique using a J-type long sheath seems to be a reasonable option when trans-femoral catheterization is difficult. PMID- 16715668 TI - MR imaging of Liliequist's membrane. AB - Liliequist's membrane is an arachnoid structure well-known to neurosurgeons. However, the importance of this membrane had been lost until the development of endoscopic third ventriculostomy (ETV). ETV is superior in its minimal invasiveness, but in some subgroups of hydrocephalus, the effectiveness of ETV may be reduced. Liliequist's membrane may block the cerebrospinal fluid (CSF) flow from the defect of the third ventricle floor, which may cause failure of ETV. Liliequist's membrane can be visualized on magnetic resonance (MR) imaging in normal healthy individuals, however, its visibility is different among individuals. CSF artifacts exist to varying degrees, but do not impede visualization of Liliequist's membrane in most subjects. Since Liliequist's membrane is a cisternal structure, the three-dimensional (3D) constructive interference in steady state (CISS) sequence is useful. The outcome of ETV could be predicted with MR imaging findings of Liliequist's membrane in a patient with obstructive hydrocephalus. High-field (> or =3 Tesla) MR imaging of Liliequist's membrane also offers superior resolution and is expected to provide additional information about Liliequist's membrane. PMID- 16715669 TI - Postoperative radiation therapy for carcinoma of the uterine cervix. AB - Postoperative radiation therapy (PORT) for cervical cancer has been empirically performed for patients with pathologic risk factors for recurrence after surgery. The efficacy of PORT is mainly supported by retrospective studies. Despite convincing evidence demonstrating a reduction in pelvic recurrence rates when PORT is employed, there is no evidence that it eventually improves patient survival. Local recurrence, such as vaginal stump recurrence, is not always fatal if diagnosed earlier. Some patients, unfortunately, may develop distant metastases even after PORT. The positive effects of PORT also may be counterbalanced by increased toxicities that result from combining local therapies. These factors obscure the efficacy of PORT for cervical cancer patients. There has been no consensus on the predictive value of risk factors for recurrence, which renders indication of PORT for early-stage cervical cancer quite variable among institutions. Today, efforts have been made to divide patients into three risk groups based on the combination of risk factors present after radical hysterectomy. In Europe/USA and Japan, however, a fundamental difference exists in the indications for radical surgery, highlighting differences in the concept of PORT; "adjuvant pelvic irradiation for stage IB-IIA patients after complete resection" in Europe/USA and "pelvic irradiation after surgery irrespective of initial clinical stage and surgical margin status" in Japan. Thus, it is questionable whether scientific evidence established in Europe/USA is applicable to Japanese clinical practice. The purpose of this article is to review the role of PORT by interpreting the results of clinical studies. PMID- 16715670 TI - Evaluation of hypoxia-specific cytotoxic bioreductive agent-sodium borocaptate 10B conjugates as 10B-carriers in boron neutron capture therapy. AB - PURPOSE: To evaluate the usefulness of 5 new 10B-compounds (TX-2091, TX-2095, TX 2097, TX-2100, and TX-2110) as 10B-carriers in boron neutron capture therpy (BNCT). They were conjugates that had been synthesized from a hypoxia-specific cytotoxic bioreductive agent, quinoxaline oxide TX-402, and a clinically used 10B carrier, sodium borocaptate-10B (BSH). MATERIALS AND METHODS: The 5 new compounds were hybrid compounds that have both a hypoxic cytotoxin unit and a thermal neutron-sensitizing unit, BSH. These new compounds and BSH were administered intraperitoneally to SCC VII tumor-bearing mice. Then, the 10B concentrations in the tumors and normal tissues were measured by gamma-ray spectrometry. Subsequently, SCC VII tumor-bearing mice were continuously given 5-bromo-2' deoxyuridine (BrdU) to label all proliferating (P) cells in the tumors, then treated with TX-2100, which was chosen based on the results of the above mentioned biodistribution analyses, or BSH in the same manner as in the biodistribution studies. Right after irradiation, during which intratumor 10B concentrations were kept at levels similar to each other, the tumors were excised, minced, and trypsinized. The tumor cell suspensions thus obtained were incubated with cytochalasin-B (a cytokinesis blocker), and the micronucleus (MN) frequency in cells without BrdU labeling [= quiescent (Q) cells] was determined using immunofluorescence staining for BrdU. Meanwhile, the MN frequency in the total (P+Q) tumor cell population was determined from the tumors that were not pretreated with BrdU. Clonogenic cell survival was also determined in mice given no BrdU. RESULTS: 10B biodistribution analyses in tumors, brain, skin, muscles, blood, and liver indicated that TX-2100 has the most favorable characteristics for concentrating a sufficient amount of 10B in tumors and maintaining a high enough 10B concentration during irradiation. In addition, TX-2100 had a significantly stronger radio-sensitizing effect with reactor thermal neutron beams than BSH on both total and Q cells in solid tumors. Further, TX-2100 clearly exhibited a radio-sensitizing effect with gamma-rays not only on total cells but also on Q and hypoxic tumor cells, which was not achieved by BSH. CONCLUSION: A 10B-carrier that acts as a hypoxic cytotoxin on tumor cells as well as having the potential to keep 10B in tumors and sensitize tumor cells more markedly than conventional 10B-carriers, such as TX-2100, is a promising candidate for use in BNCT. PMID- 16715672 TI - Chemoradiation therapy for cervical cancer: toxicity of concurrent weekly cisplatin. AB - PURPOSE: To retrospectively evaluate the toxicity of concurrent weekly cisplatin and radiation therapy (RT) for locally advanced cervical cancer. MATERIALS AND METHODS: Between April 2001 and December 2004, 21 consecutive previously untreated patients with locally advanced cervical cancer were treated with concurrent chemoradiation therapy (CCRT) at the Tokushima University Hospital. Clinical stages were II: 5, III: 15, IVA: 1. External beam radiation therapy (EBRT) was delivered with 10 MV X-rays, 2 Gy fraction per day; total dose to the whole pelvis was 50 Gy. Iridium-192 high-dose-rate (HDR) intracavitary radiation therapy was performed with 10-30 Gy (median, 24 Gy) targeted at point A. Concurrent chemotherapy consisted of cisplatin, administered weekly at a dose of 40 mg/m2 for patients who were younger than 65 years and 30 mg/m2 for those 65 years or over. A maximum single dose of cisplatin, up to 70 mg/body, was administered in 5 cycles during EBRT. RESULTS: A total of 86 cycles of cisplatin were administered to the 21 patients, with a median of 4 cycles (range, 2-5). Severe hematological toxicity occurred in 18 patients (86%), including grade 3 in 17 patients (81%) and grade 4 in one patient (4.8%). Moderate or severe gastrointestinal toxicity occurred in 11 patients (52%), including grade 2 in 10 patients (48%) and grade 3 in one patient (4.8%). The grades of hematological toxicity were significantly greater in the 40 mg/m2 group than in the 30 mg/m2 group. All of the patients who were administered 40 mg/m2 of cisplatin developed grade 3 or greater hematological toxicity, including one patient with grade 4 toxicity. In the 30 mg/m2 group, 3 of 10 patients developed less than grade 3 toxicity, and all patients completed radiation therapy without interruption. CONCLUSION: The incidence of severe acute hematological toxicity was significantly higher in this study than in previously reported randomized controlled trials (RCTs), especially in the group of 40 mg/m2 cisplatin. A dose of 30 mg/m2 of cisplatin was considered to be feasible in weekly cisplatin and radiation therapy. PMID- 16715671 TI - Detection of intraductal component around invasive breast cancer using ultrasound: correlation with MRI and histopathological findings. AB - PURPOSE: The purpose of this study was preoperatively to diagnose the intraductal component, which is indispensable in planning for breast conservation therapy, and also to minimize local recurrence. This study investigated the efficacy of ultrasound (US) in the detection of intraductal component in comparison with magnetic resonance imaging (MRI) and histopathological findings. PATIENTS AND METHODS: In 47 patients with invasive breast cancer, US features of the intraductal component were classified as (a) solid ductal dilatation radiating from the tumor, (b) presence of satellite lesion in the same segment without ductal dilatation, and (c) ductal dilatation between the main tumor and satellite lesion. MRI depicted intraductal extension as the most enhanced area during the first or second phase of the dynamic study. Other criteria for the detection of the intraductal component by MRI were as follows: (a) a satellite lesion around the main tumor, (b) bridging enhancement between the main tumor and satellite lesions. The extent of the intraductal component was measured and classified as mimimal (0-5 mm), moderate (6-15 mm) or wide (>15 mm). RESULTS: In 28 of 47 (60.0%) patients, a wide intraductal component of more than 15 mm was proved histopathologically. Of 28 patients, US and MRI could accurately detect wide intraductal components in 16 patients and 14 patients, respectively. Sensitivity, specificity, and accuracy were 57.1%, 84.2%, and 68.1% respectively for US and 50.0%, 89.5% and 65.9% for MRI, respectively. When both US and MRI results were combined, sensitivity, specificity, and accuracy were 75.0%, 84.2%, and 78.7%. CONCLUSION: Current US examination depicted the intraductal component of breast cancer more accurately than MRI. Further, our study suggests that the use of both US and MRI together is complementary and offers more advantage than US alone. PMID- 16715673 TI - Change in dose distribution of three-dimensional conformal radiotherapy during treatment for lung tumor. AB - PURPOSE: We investigated the changes in dose distribution of three-dimensional conformal radiotherapy (3D CRT) during lung tumor treatment. MATERIALS AND METHODS: Ten patients with non-small cell lung cancer who had undergone planning for radical radiotherapy were selected for study. Computed tomography (CT) examination was performed at two time intervals during the course of conformal radiotherapy: t0 Gy at the time of planning and t40 Gy at 40 Gy of treatment. We transferred all the planned beam data at t0 Gy to each t40 Gy CT image. The isodose distribution was recalculated at time t40 Gy for the same beam characteristics. Variations in volumes and dose-volume histograms (DVHs) were analyzed and compared for lung, gross target volume (GTV), and planning target volumes (PTV) between t0 Gy and t40 Gy. A paired t-test was performed to compare the DVH between t0 Gy and t40 Gy. RESULTS: The mean minimum doses for t40 y GTV, and PTV were lower than t0 y. However, there was no significant difference between t0 Gy and t40 Gy (p=0.493, 0.378, respectively). There was a patient whose minimum doses of GTV and PTV were decreased and who had notable improvement of lobar atelectasis after 40 Gy of radiotherapy. Comparison of the percent volume of received dose exceeding 20 Gy (V20) and the mean dose for the total lung revealed that t40 Gy was larger than to Gy (p=0.013, 0.012). CONCLUSION: Incorporation of the time factor into 3D treatment planning is mandatory for frequent reiteration of treatment planning during treatment periods. Clearly, more work in this area should be considered. PMID- 16715674 TI - Comparative study of vascular enhancement on post-contrast CT using three dosages of iodinated contrast media for the aim of detecting brain metastasis in patients with lung cancer. AB - OBJECTIVE: A prospective double-blind randomized study was performed to compare the contrast of vascular enhancement using three dosages of iodinated contrast media for a possible metastatic lesion in the brain. MATERIALS AND METHODS: Sixty six patients with lung cancer received brain computed tomography (CT) with intravenous administration of iodinated contrast medium (CM). The patients were randomly assigned to receive one of the three types of CM: 30 g iodine, 24 g iodine, and 15 g iodine. Three radiologists judged the degree of vascular contrast enhancement and diagnosed the presence of brain metastasis. The CT numbers in major arteries were also measured. RESULTS: The subjective average scores with standard deviation were 2.06+/-0.48, 1.97+/-048, and 1.44+/-0.43, and the measured average CT numbers with standard deviation (SD) were 168.5+/-39.6, 166.1+/-28.6, and 146.1+/-27.0 HU with 30 g, 24 g, and 15 g iodine, respectively. The scores and the CT numbers in 15 g iodine were less than those with 30 g and 24 g iodine. Brain metastasis was detected in one patient each in groups A and C, and one false-positive case was found in group B. CONCLUSION: CT study with a dose of 24 g iodine showed equivalent quality on vascular enhancement in comparison with a 30 g iodine dose. PMID- 16715675 TI - Preliminary study of correction of original metal artifacts due to 1-125 seeds in postimplant dosimetry for prostate permanent implant brachytherapy. AB - PURPOSE: We investigated a subtraction-based reprojection approach to reduce CT metal artifacts due to I-125 seeds and evaluated the clinical implications in postimplant dosimetry for prostate permanent implant brachytherapy. MATERIALS AND METHODS: The raw projection data were used to reduce metal artifacts due to I-125 seeds. CT images of the metal parts only were separated from the original CT images by setting the threshold for pixel value to that of the I-125 seeds. Using these images, sinograms of CT images with and without seeds were obtained by inverse Radon transform (iRT), and the sinogram of the metal image was subtracted from that of the original image. Finally, the image was reconstructed using the sinogram by Radon transform (RT). This technique was applied to a prostate phantom and to a patient undergoing prostate permanent implant brachytherapy. RESULTS: Metal artifacts from I-125 seeds were reduced in both the phantom and patient studies. This technique decreased the density of the inner region of seeds but enhanced the density of the seed edge, thereby facilitating the identification of seed number, orientation, and location. CONCLUSION: This method reduces metal artifacts from I-125 seeds, and has potential for decreasing the time required for and improving the accuracy of postimplant dosimetry. PMID- 16715676 TI - A case of spontaneous regression of hepatocellular carcinoma with multiple lung metastases. AB - Spontaneous regression of hepatocellular carcinoma (HCC) is a rare phenomenon. We present herein the case of a patient with hepatocellular carcinoma with multiple lung metastases in whom malignancy spontaneously regressed after taking Pheliinus linteus Mycelium. A 79-year-old man consulted our hospital complaining of epigastric discomfort. Abdominal MRI and CT revealed a 3 cm diameter tumor in the liver, and chest CT showed numerous nodular lesions. The levels of alpha fetoprotein (AFP) and protein induced by vitamin K deficiency or antagonist-II (PIVKA-II) were very high. We diagnosed HCC with multiple lung metastases, and no therapy was performed. Independently he took exact from Phellinus linteus Mycelium for one month, and 6 months later the tumors appeared to be in complete regression. The mechanism underlying this intriguing phenomenon remains unknown. PMID- 16715677 TI - Non-necrotic invasive squamous cell carcinoma associated with an inverted papilloma: MRI features. AB - We present the MRI findings of a carcinoma associated with an inverted papilloma located in the nasal cavity. This tumor can be divided into 2 types, those with and those without gross central necrosis. In our case, the tumor did not have any gross central necrosis, and MRI showed convoluted cerebriform patterns. We found that MRI is unable to distinguish malignant and benign regions in a central mass with no gross necrosis, since this lesion does not alter the basic morphological pattern of inverted papilloma. PMID- 16715678 TI - Postmortem computed tomographic (PMCT) and postmortem magnetic resonance imaging (PMMRI) demonstration of fatal massive retroperitoneal hemorrhage caused by abdominal aortic aneurysm (AAA) rupture. AB - We report a case of fatal massive retroperitoneal hemorrhage caused by the rupture of an abdominal aortic aneurysm (AAA) in which postmortem computed tomography (PMCT) and postmortem magnetic resonance imaging (PMMRI) provided clear delineation. In this case, the autopsy imaging system using PMCT and PMMRI was useful as a screening method to determine the cause of death as AAA rupture and became a guide for the subsequent autopsy. PMID- 16715679 TI - Skeletal muscle: an unusual site of distant metastasis in gastric carcinoma. AB - Gastric cancer metastatic to skeletal muscle is an unusual entity. Surgery, systemic chemotherapy, or radiotherapy to the metastatic mass can be treatment options for achiving palliation. CASE REPORT: A patient with multiple skeletal muscle metastases that occurred during follow-up after gastrectomy and adjuvant chemo-radiotherapy is reported. Magnetic resonance imaging (MRI) demonstrated soft-tissue masses involving the posterior right paralumbar and posterior left paradorsal muscles. Biopsy showed metastatic infiltrating adenocarcinoma. The patient did not respond to palliative chemotherapy. Palliative radiotherapy was administered to the painful mass. Based on this case, the diagnosis of muscle metastases and treatment options for palliation are discussed. PMID- 16715680 TI - Bronchial granular cell tumor: a case presenting secondary obstructive changes on CT. AB - Bronchial granular cell tumor is an uncommon benign tumor that causes pulmonary complications due to obstruction of the airways. We report a case presenting bronchial obstruction on high resolution (HR) computed tomography (CT). PMID- 16715681 TI - Issues in the study of intonation in language varieties. AB - Some key issues in the study of intonation in language varieties are presented and discussed with reference to recent research on the intonation of New Zealand English. The particular issues that are highlighted include the determination of the intonational phonological categories of a language variety, and the attribution of varietal differences as realizational differences between varieties or as systemic differences in the categories found to be present in each variety. PMID- 16715682 TI - Pitch accent alignment in romance: primary and secondary associations with metrical structure. AB - The article describes the contrastive possibilities of alignment of high accents in three Romance varieties, namely, Central Catalan, Neapolitan Italian, and Pisa Italian. The Romance languages analyzed in this article provide crucial evidence that small differences in alignment in rising accents should be encoded phonologically. To account for such facts within the AM model, the article develops the notion of "phonological anchoring" as an extension of the concept of secondary association originally proposed by Pierrehumbert and Beckman (1988), and later adopted by Grice (1995), Grice, Ladd, and Arvaniti (2000), and others to explain the behavior of edge tones. The Romance data represent evidence that not only peripheral edge tones seek secondary associations. We claim that the phonological representation of pitch accents should include two independent mechanisms to encode alignment properties with metrical structure: (1) encoding of the primary phonological association (or affiliation) between the tone and its tone-bearing unit; and (2), for some specific cases, encoding of the secondary phonological anchoring of tones to prosodic edges (moras, syllables, and prosodic words). The Romance data described in the article provide crucial evidence of mora-edge, syllable-edge, and word-edge H tonal associations. PMID- 16715683 TI - Microvariation in accentual alignment in Basque Spanish. AB - This paper presents patterns of accentual alignment in two varieties of Spanish spoken in the Basque Country: Lekeitio Spanish (LS), with speakers whose other native language is Lekeitio Basque (LB); and Vitoria Spanish (VS), with monolingual speakers of Spanish from the city of Vitoria. These patterns are compared to those of Madrid Spanish (MS), compare Face (2002). In LS, accents are realized as pitch rises rather than falls, like in MS and unlike in LB, but peaks are aligned before the offset of the accented syllable, unlike in MS and like in LB. At the end of the subject phrase, peaks display later alignment, like in MS. Thus, LS displays mixed properties of Basque and Spanish intonation. In VS, stress is also realized as a tonal rise, with peaks aligned after the offset of the accented syllable, like in other varieties of Spanish and unlike LS. The low tone target is aligned before the onset of the stressed syllable, earlier than in LS and MS. The continuum of tonal target alignment is observed in LS, VS, and MS, and the difficulties in identifying a "starred" tone lead to a discussion of the suitability of the starred tone notation. PMID- 16715684 TI - Tonal alignment in Irish dialects. AB - A comparison of the contour alignment of nuclear and initial prenuclear accents was carried out for the Irish dialects of Gaoth Dobhair in Ulster (GD-U) and Cois Fharraige in Connaught (CF-C). This was done across conditions where the number of unstressed syllables following the nuclear and preceding the initial prenuclear accents was varied from 2-0. This tests a variable peak hypothesis prompted by findings for other languages, that peak timing drifts as a function of the number of syllables preceding (the prenuclear) and following (the nuclear) accent. These data also test a second hypothesis that the L*+H dominant accent of GD-U might be viewed as being underlyingly the same as the dominant H* or H*+L accent of the CF-C dialect. According to this realignment hypothesis, the difference between these Ulster and Connaught dialects lies in the way that the melodic tier is aligned to the segmental tier: GD-U would be viewed as having a delayed realization of the peak relative to the Connaught dialect. Results do not support the variable peak hypothesis for Irish, as in either dialect, the peak appeared to be rather fixed across the three conditions examined (though not necessarily identical for prenuclear and nuclear positions). The results also militate against the realignment hypothesis, which rather than providing a more simple unifying account, would greatly complicate it. One reason is that there is a peak timing difference between the nuclear and prenuclear accents of CF-C, not mirrored in GD-U. Furthermore, even if one were to limit consideration to a single (e.g., prenuclear) context, a simple realignment of the accents in one dialect does not generate the appropriate contour in the other. PMID- 16715685 TI - The mouse footpad model. PMID- 16715686 TI - The mouse foot-pad technique for cultivation of Mycobacterium leprae. AB - Although multiplication of Mycobacterium leprae in the foot pads of immune competent mice is limited, and no leprosy-like lesions are produced in these animals, the mouse foot-pad system represents the first truly useful and reproducible animal model of M. leprae infection. Its employment has enabled research into basic questions with respect to the microbiology of M. leprae, and the epidemiology, treatment and control of leprosy. The mouse foot-pad technique is labour-intensive and time-consuming, and is expensive in terms of the costs of animal purchase and maintenance. In addition, the technique appears to be rather imprecise and insensitive, compared with the techniques employed in working with cultivable micro-organisms. For these reasons, and also as a by-product of the success of multi-drug therapy, the technique has been abandoned in many research centres. Nevertheless, until a more simple and sensitive technique for demonstrating the viability of M. leprae is developed, the mouse foot-pad system remains an essential tool for leprosy research. In this review, we discuss the mouse foot-pad technique in detail, analyse its precision, point out its shortcomings, describe its most important applications, and prescribe a method by which to assess the ability of an alternative technique to serve in place of this established technique. PMID- 16715687 TI - Multi-centre, double blind, randomized trial of three steroid regimens in the treatment of type-1 reactions in leprosy. AB - OBJECTIVE: The objective of this randomized trial was to compare three different steroid regimens in treating type 1 reactions in leprosy in routine clinical practice. DESIGN: The study design was a multicentre, double-blind, randomized, controlled, parallel group trial in patients with acute reversal reactions. The trial was conducted in six leprosy treatment centres in India. A total of 334 participants with acute type 1 reaction were recruited to the trial and randomized to one of three prednisolone regimens: high dose (60 mg per day) or low dose (30 mg per day) both tapered over 20 weeks, and short duration (60 mg per day tapered over 12 weeks). The main outcome measure was the proportion of patients failing to respond to treatment and requiring additional steroids. RESULTS: At the end of 12 months, 46% on the short course required additional steroids compared with 31% on the low dose and 24% on the high dose regimen. CONCLUSIONS: The two 20-week regimens were significantly better than the 12-week regimen. The high dose 20-week regimen was marginally and non-significantly better than the low dose regimen, but the high dose regimen contained 50% more steroid. Reactions in leprosy persist over many months and require long courses of steroids. PMID- 16715688 TI - Characteristics of known leprosy contact in a high endemic area in Brazil. AB - BACKGROUND AND PURPOSE: The annual number of new cases of leprosy has not declined in Brazil over the last 15 years, indicating that transmission continues at the same level. To study transmission, we interviewed leprosy patients about their known leprosy contact (KLC). METHODS: Clinical and demographic data were collected from 506 leprosy patients in four health units in the Metropolitan Region of Vitria, State of Espirito Santo, Brazil. SPSS 9.0 was used as a database and analysis. RESULTS: Two hundred and twenty-six (44.7%) of 506 leprosy patients reported KLC, 136 (60.2%) of 226 were parents. Among 226, the mean of KLC was 1.89 (SD +/- 1.65), and 61.3% had one KLC. KLC as a household contact was reported by 92 (40.7%) out of 226, and 121 (53.5%) had no household contact. KLC were most frequently sisters and brothers in the PB cases, and sons/daughters in MB cases. Mothers occurred more frequently as a KLC than fathers. From the leprosy patients that had reported household contacts, 73% said that at the onset of their skin lesions, the KLCs were either undergoing were not yet released from treatment (RFT), and 23.45% had not begun the treatment yet. Altogether, 62.3% of 226 cases had daily contact with the KLC. CONCLUSION: In Brazil, household contacts, including the family members (mothers, sisters and brothers), as well as the social contact need to be investigated by the control programs. PMID- 16715689 TI - Delay in the diagnosis of leprosy in the Metropolitan Region of Vitoria, Brazil. AB - This paper reports on the time between the onset of the first lesion and diagnosis, defined as delay, and is based on results obtained by interviewers from a survey carried out amongst 450 leprosy patients in a leprosy endemic area in the Metropolitan Region of Vitoria (MRV), state of Espirito Santo, Brazil. The mean age at diagnosis in all cases was 41.47 years and the median was 42.5 years. The mean age at diagnosis in MB (42.9 years) was greater than in PB (38.5 years). The mean of the delay in all cases was 25.25 months, median 12 months and range 0 360 months. The mean of the delay in MB (27.2 months) was greater than in PB (21.3 months). The results of this study suggest that although the delay in leprosy diagnosis in this region of Brazil was not too long when it was compared with other studies in endemic countries, it is still a problem: 65.4% of patients were diagnosed after a delay of 6 months. The Leprosy Control Programme in this state needs more effective health education in order to reduce the current period of delay before diagnosis. PMID- 16715691 TI - Knowledge of and attitudes to leprosy among patients and community members: a comparative study in Uttar Pradesh, India. AB - The roles of literacy and gender in enhancing help seeking behaviour in leprosy need further research in order to maximize the effectiveness of health education programmes. A study on leprosy knowledge and attitudes was carried out in Uttar Pradesh, one of the hyper endemic states for leprosy in north India, on a random sample of 130 leprosy patients, 120 non-leprosy patients, and 150 community members. A questionnaire was prepared, tested and administered in Hindi, the local language, by a qualified interviewer. Statistical analyses were done in each group by gender and literacy, and compared. Almost everyone in the three groups knew of leprosy, but only a larger proportion of leprosy patients (60%) mentioned anaesthetic patch, as compared to about 20% or less in the other groups. A vast majority in all groups mentioned bad blood, or divine curse as the cause. Even among leprosy patients, less than 10% of illiterates and only about 40% of literates cited infection as the cause of leprosy. Literates had a better, though still quite a poor knowledge on the symptoms as well as the causation of leprosy. However, almost all stated that leprosy was curable, though they couldn't mention MDT specifically. They felt that not all patients need have deformity. About 20-30% of the leprosy affected, but nearly 50-60% in the other groups stated that there was discrimination. Nearly 70% felt that leprosy affected social participation, over 90% attributing this to adverse social stigma. Multivariate analyses, adjusted for sex, confirmed the significant association of literacy with both knowledge and attitudes. In the light of massive health education and IEC campaigns, the findings from this study are disappointing. Adult literacy programmes combined with more innovative focused approaches to suit various target audiences can impact knowledge and attitudes better. PMID- 16715690 TI - Risk factors for developing leprosy--a population-based cohort study in Indonesia. AB - We identified risk factors associated with increased yearly incidence rates of leprosy in five island populations. Age, sex, household size and Mycobacterium leprae-specific antibodies as well as contact factors were studied. Of 94 index patients (patients diagnosed in 2000), 43 (46%) were classified as multibacillary (MB), 17 (19%) were seropositive for PGL-1 [corrected] antibodies and 6 (7%) had M. leprae DNA in nasal swabs as determined by polymerase chain reaction (PCR) testing. All PCR positive patients were also seropositive. Forty-four of 4903 initially symptom free persons developed leprosy within 4 years, giving an incidence rate of 298 per 1000 person-years. Men had a 22 times higher risk [95% confidence interval (CI): 1.2-4.1] of developing leprosy than women. People living in households with more than 7 members had a 3.1 times higher risk (95% CI: 1.3-7.3) than households of 1-4 members. Persons who were seropositive in 2000 had a 3.8 times higher risk (95% CI: 1.1-12.6) than seronegative persons. Household contacts of MB patients had an adjusted hazard ratio (aHR) of 4.6 (95% CI: 1.6-12.9) and household contacts of PCR positive patients an aHR of 9.36 (95% CI: 2.5-34.9) compared with non-contacts. Patients with PCR positive nasal swabs, suggesting nasal excretion of M. leprae, are probably the patients with the highest transmission potential. Since all index patients who were PCR positive were also seropositive, serology seems an adequate tool to identify these patients. Preventing seropositive persons from becoming seropositive and infectious patients might break the chain of transmission. PMID- 16715692 TI - Reverse medial plantar artery flap. PMID- 16715693 TI - Immune reconstitution inflammatory syndrome in an HIV seropositive leprosy patient. PMID- 16715694 TI - Bigger may not be better. PMID- 16715695 TI - Discrimination against leprosy patients and their families: the situation (early 2005) in West Darfur, Sudan. PMID- 16715696 TI - Media perception of leprosy. PMID- 16715697 TI - [Taxanes in neojuvant radio-, chemoradio- and chemotherapy for breast cancer]. PMID- 16715698 TI - [Neojuvant radio-, chemoradio- and chemotherapy for locally advanced stomach cancer]. PMID- 16715699 TI - [Impact of chemical carcinogens on amphibia]. PMID- 16715700 TI - [Combined radiotherapy for inoperable esophageal cancer]. AB - Among 112 patients with inoperable esophageal cancer treated with high-dose intraluminal brachytherapy and distant irradiation, objective response was in 84.8%. Predominantly local complications were reported in 78.6%. Recurrences were detected in 63.5% during follow-up, with mean relapse-free period being 107 days. Mean survival time was 15.3 months (1-, 2- and 3-year survival--58.8; 22.3 and 12.9%, respectively) which was comparable to similar results of palliative esophagectomy for gastric cancer. PMID- 16715701 TI - [A theoretical model for assessing the relationship between "stage migration" and survival of gastric cancer patients after variable lymph node dissection]. AB - Standard histological procedures identify metastatic gastric cancer in approximately 15% of cases of macroscopically unaltered lymph nodes. Such metastases evade detection after surgery due to inadequate lymph node dissection, thus increasing staging error. Consequently, about 15% of patients suffer from tumor which is actually more advanced than one diagnosed originally. PMID- 16715702 TI - [Familial well-differentiated thyroid carcinoma]. AB - Medical Research Institute of Radiology, Russian Academy Forty-eight cases of familial disease (24 families) (4.3%) were identified among 1,118 patients with well-differentiated thyroid carcinoma who had been either examined or treated at the Clinic of Medical Research Institute of Radiology (1995-2004). In 86% of the study group, papillary thyroid carcinoma (PTC) was associated with tumor of the identical histological pattern while the remaining families revealed association with follicular or medullary thyroid cancer. Carcinoma inheritable from mother was the most frequent (75%). No differences in manifestation, histological pattern, stage or clinical course were established following a detailed evaluation of clinico-morphological data on 43 familial and 172 sporadic (control) cases in both groups. The analysis pointed to a significantly higher incidence of concomitant thyroid pathology in the familial thyroid cancer group. Molecular-genetic study of RET-protooncogene and gene BRAF in 6 blood samples from PTC-bearers established RET-mutation (mother and daughter) in codon 891 (exon 15) G2673A (TCG->TCA). No mutation in BRAF was found. PMID- 16715703 TI - [Poorly-differentiated thyroid carcinoma: morphological verification and differential diagnosis]. AB - Poorly-differentiated thyroid carcinoma is a stage in the development of anaplastic carcinoma from well-differentiated neoplastic transformation of follicular epithelium. Such tumors differ with respect to histological pattern and age. For instance, patients, aged 50 and more, revealed great nodular size (50-70 mm), marked invasion, large cells with high mitotic index and solid and/or insular pattern. In the group under 40, incapsulated poorly-differentiated tumors were more frequent; they had trabecular histological pattern alone with a much smaller size (ca. 30mm). Further clinico-morphological studies might solve the problem of the connection between cyto- and histological pattern, on the one hand, and clinical course and prognosis, on the other. PMID- 16715704 TI - [Prognostic significance of tumor cell ploidy in advanced ovarian carcinoma]. AB - A micro-imaging retrospective investigation of tumor cell ploidy using exudates smears from 52 patients with serous ovarian adenocarcinoma (stage III-IV) showed survival in those with diploid tumor to be significantly longer (38 months) than in cases of aneuploid malignancies (16 months). Significant differences in survival versus ploidy, stage, residual tumor size, therapy modality and age were established with the aid of univariate analysis (Caplan-Meyer) of 5-year survival. Multivariate step-by-step regressive analysis (Cox) was carried out on the prognostic significance of such factors as residual tumor size (p=0.013), stage (p=0.019) and tumor ploidy (p=0.035). Further study showed ploidy to be a fully independent factor like any other factors of ovarian carcinoma development. PMID- 16715706 TI - [Effect of preliminary irradiation on DNA-ase activity and DNA fragmentation degree in the nuclei of Guerin's carcinoma cells]. AB - The effect of preliminary irradiation on DNA-ase activity and DNA fragmentation degree in the nuclei of Guerin's carcinoma was evaluated. It was found that effective growth of malignant cells involved higher concentrations of acid DNA ase matched by decreased levels of alkaline one which in turn lowered the rates of DNA fragmentation. Low-dose preliminary irradiation stimulated relevant processes. PMID- 16715705 TI - [The effect of preliminary administration of water with reduced deuterium content on the growth of transplantable tumors in mice]. AB - It was investigated whether preliminary administration of water with reduced Deuterium content may modify the inhibitory effect of the water given to BDF1 and CDA mice on the day of tumor transplantation. Two models were used: Lewis lung carcinoma (LLC) and uterine cervical carcinoma (UCC). Experimental mice (20 per group) used the water with reduced Deuterium content. Water with a Deuterium content close to that of drinking water was given to controls (30 per group). Both kinds of water were given to mice 4 weeks before the experiment. Inhibitory effect was acknowledged by the time at which the first nodules appeared at site of transplantation and assessed with respect to volume of tumor and animal life span. Metastasis inhibition coefficient was determined for Lewis carcinoma metastasizing to the lung. It was found that water with reduced Deuterium content effectively inhibited the growth of the transplantable tumors under study, significantly reduced metastasis weight in mice with LLC and increased life-span in animals with transplantable UCC. PMID- 16715707 TI - [Effect of exogenous DNA on the growth of transplantable tumors]. AB - Using transplantable Ehrlich ascites tumor, hepatoma HA-1 and Lewis carcinoma it was shown, that preparations of fragmented genomic DNA can more or less effectively inhibit such tumors as well as the growth of their metastases. Such effects were produced by DNA preparations derived from tissues of mice, both syngeneic or allogeneic to tumor-bearer, as well as from human tissues. PMID- 16715709 TI - [Methodologic and organizational principles of selective screening for cervical, endometrial and ovarian carcinoma]. AB - Data are presented on screening for cervical (CC), endometrial (UC) and ovarian (OC) carcinoma carried out in the Republic of Belarus as well as a discussion of logistic and methodologic aspects of the problem. Local "centers for prevention and early diagnosis of reproductive organ tumors" form the backbone of the national system. They operate on the basis of the existing guidelines about such matters as screening staging, sites, examination procedures used at each stage and suitable distribution of medical personnel (obstetricians, doctor's assistants, gynecologists). A comparison of data on selective screening for CC, UC and OC with those of mass prophylactic examinations and cytological screening for CC showed that selective detection rates for cervical dysplasia were 14 times, and in situ cervical carcinoma--4 times higher. That was followed by lower incidence of invasive CC and absence of advanced cases. Consequently, the share of endometrial hyperplasia cases rose 3.5-fold, uterine myoma--3-fold and adenomyosoma--1.5-fold. identified atypical endometrial hyperplasia, in situ cancer and endometrial stage I in "clinically healthy" women. The rates of detection of such tumor-like ovarian formations as follicular, lutein, endometrioid and para-ovarian cysts increased 2.3-fold, benign tumors--2.2-fold and polycystosis--twofold. PMID- 16715708 TI - [Comparison of the effectiveness of mono- and polychemotherapy for murine lymphosarcoma, resistant to apoptosis induction]. AB - Strain-specific murine lymphosarcoma RLS, resistant to apoptosis induction, was used to demonstrate that polychemotherapy (cyclophosphamide+adriamycin+cisplatin) failed to gain any significant advantage over monotherapy with adequate doses of those drugs PMID- 16715710 TI - [Organ-saving and function-conserving procedures in the treatment of cervical carcinoma]. AB - The paper presents our data on conservative treatment of early cervical carcinoma using laser, ultrasound, electrocautery scissors and electrosurgical conization. Also, menstrual and reproductive function in cured patients as well as frequency of tumor recurrence are discussed. An original procedure for ovarian transposition used in combined treatment of invasive cervical carcinoma is suggested. Potential and methods of monitoring transposed appendages are validated. Findings on 5-year survival are presented. The methods of conservative treatment are evaluated as promising further achievements in oncogynecology. PMID- 16715711 TI - [Improvement of postoperative radiotherapy in the complex treatment of stage II III cervical cancer]. AB - Clinical data on the management of squamous-cell cervical carcinoma stage II-III FIGO are presented. At stage I, treatment included neoadjuvant polychemotherapy (NACT)--cisplatin 100 mg/m2, carboplatin 450 mg/m2 in conjunction with 5-FU 250 mg/m2 or paclitaxel 135 mg/m2. According to protocol, two identical courses of NACT were given, at 21-28 day interval, followed by extended extirpation of the uterus (Wertheim). Immediate results and complications caused by preoperative radiotherapy, important for postoperative one, were evaluated, concepts and strategies for postoperative intracavitary and distant radiotherapy worked out. To optimize radiotherapy, dosage was reduced by 10% or daily single target dose of intracavitary and distant irradiation split per 24 hrs, indications for preoperative radiotherapy variants were extended, and local dosage escalation in areas of lymphocyst or hematoma renounced. Overall survival was 85%, recurrence free--77.5%. PMID- 16715712 TI - [Treatment outcomes in patients with unfavorable histologic type of endometrial carcinoma]. AB - Five-year overall and recurrence-free survival was evaluated in 84 stage I endometrial cancer patients with such histological patterns as poorly differentiated endometrioid adenocarcinoma, clear-cell carcinoma, serous papillary cancer, poorly differentiated cancer and mixed-type carcinoma. Overall 5-year survival in the whole group was 60.1 +/- 5.9% while recurrence-free survival--52.1 +/- 5.9%, those rates for poorly differentiated endometrioid adenocarcinoma being 50.8 +/- 7.3% and 41.9 +/- 7.4%, respectively. Our data suggest that combination therapy is ineffective in the management of stage I endometrial carcinoma when histotype of tumor is unfavorable (overall 5-year survival - 56.51 +/- 7.8%; recurrence-free survival--49.7 +/- 7.9%). PMID- 16715713 TI - [Proton irradiation of the pituitary gland for alleviating pain in patients with disseminated prostate cancer]. AB - Central Research Institute of Roentgeno-Radiology; Medical Academy for Further Education, St. Petersburg Stereotactic ablation of the frontal lobe of the pituitary with narrow beams of 1,000 MeV protons was performed in 80 patients to alleviate pain caused by bone metastases. Pain was aborted for a long time so that pain-relieving medication was suspended in 56 (70%) patients. The remaining patients cut down on taking analgetics: peripheral drugs were taken by 11(13.75%), non-narcotic opioids (tramal)--4 (5%), class III opioid narcotics--7 (8.75%), and morphine-type drugs--only 2. No untoward side-effects were reported. PMID- 16715714 TI - [Epidemiology of malignant melanoma in the Republic of Tajikistan]. AB - 447 patients suffering primary malignant melanoma were registered in the Republic of Tajikistan in 1970-2000. The 1970-1974 morbidity rate of 0.33 rose to 0.51 in 1980-1984 and later fell to 0.07 (1995-2000). Primary tumor localizations were: skin (80.8%), eye (10.9%), and anorectal area (1.8%); primary focus was not identified in 2.9%. A distinction in clinical course was established between native and non-native population. Overall 5-year survival was 56.3%. PMID- 16715715 TI - [Surgical treatment of primary multiple synchronous cancer in the aerodigestive tract]. PMID- 16715717 TI - [Combination of cancer and tuberculosis of the lung]. PMID- 16715716 TI - [Myeloid sarcoma of the uterine cervix as presentation of acute myeloid leukemia]. PMID- 16715718 TI - [Limitations and inconsistencies of current criteria for lung cancer staging]. PMID- 16715719 TI - [Potential use of exogenous phosphocreatine (neoton) in the combined therapy of cancer complicated with cardiovascular diseases]. PMID- 16715720 TI - [Professor Luarsab Kalistratovich Sharashidze, founder and first director of the Oncology Research Center in Georgia]. PMID- 16715721 TI - [Extent of surgery in well-differentiated thyroid cancer]. PMID- 16715722 TI - Four decades Medicaid: reinvention, redesign and reformation needed now. PMID- 16715724 TI - Medicaid rebalancing. PMID- 16715723 TI - Trends in 2006 budget law: Federal Medicaid spending cut, but door open for home and community-based care. PMID- 16715725 TI - Medicaid fraud time to sound the alarm? PMID- 16715726 TI - The Carolinas are calling! Making the case for telehomecare! PMID- 16715727 TI - Florida's Medicaid reform proposal. PMID- 16715728 TI - Increasing needs in New Mexico. PMID- 16715729 TI - Long-term living efforts in PA stalled. PMID- 16715730 TI - Medicaid: thumbs up, or thumbs down. PMID- 16715731 TI - Network contracting and the antitrust laws: giving "messenger models" a tune-up. PMID- 16715732 TI - The "STATE" of home care and hospice. AB - There is at least one home care and hospice industry association in each of our 50 United States and Puerto Rico. These associations represent tens of thousands of home care and hospice providers. They provide state and local leadership, professional and para-professional training, and other services designed to help home care and hospice providers care for our nation's sick, disabled and elderly. These are their stories. PMID- 16715733 TI - MedPAC reviewing Medicare Hospice Benefit payment system: will report to Congress in June. PMID- 16715734 TI - States turn Medicaid towards technology and homecare. PMID- 16715735 TI - Potpourri. PMID- 16715736 TI - Startup, acquisition, or new programs and services: a framework for choosing the best business expansion strategies. PMID- 16715737 TI - Build your marketing and referral dashboard. PMID- 16715738 TI - Sustaining our calling and cause. PMID- 16715739 TI - Medicaid: mending holes in the health care safety net. PMID- 16715740 TI - Read our hands. PMID- 16715742 TI - Happy days. PMID- 16715741 TI - No easy answers. PMID- 16715743 TI - The right to choose. PMID- 16715744 TI - The courts have already delivered what the government wants from the Mental Health Act. PMID- 16715745 TI - When you're far away you realise the essence of what you truly love about people. Interview by Suzy Johnston. PMID- 16715746 TI - Participation--why bother? PMID- 16715747 TI - Giving up the habit. PMID- 16715748 TI - Acting out, acting in. PMID- 16715751 TI - Becoming a school nurse. PMID- 16715750 TI - Hormonal contraception: recent advances. PMID- 16715752 TI - Nutrition and the prevention of cancer. AB - Many studies have shown an association between nutrition and cancer and, since one in three people in England will develop cancer and one in four will die from the disease, there is great interest in prevention through diet. The issues are complex and evidence is often limited or equivocal. This review considers the strength of the evidence and puts the risks of cancer in context. It concludes that although no dietary changes can be guaranteed to prevent cancer, on balance there is enough evidence to recommend the current official "healthy eating" guidelines as the best advice for reducing the risk of diet-associated cancers. The greatest protection against cancer coincides with eating more fruit, vegetables and wholegrain foods. Overindulgence in alcohol, smoking, obesity and lack of exercise may interact with dietary factors to increase the risk of some cancers. Healthy lifestyle measures such as regular exercise, avoiding overweight, not smoking and limiting the intake of alcohol can therefore also be recommended as helping to reduce the risk of cancer. When advising individuals, it is necessary to consider all aspects of the person's lifestyle and to bear in mind that there are some limitations to the current evidence. A summary of the practical recommendations for reducing cancer risk is included. PMID- 16715753 TI - Infant nutrition--from professional to personal. AB - The author, an experienced paediatric dietitian, describes what it was like breast-feeding and weaning her first baby and how she has modified her previous professional views of infant feeding in the light of her own experience. As a result, she makes some pragmatic recommendations for helping new mothers. She urges health professionals to be more realistic about the difficulties as well as the rewards of breast-feeding and to provide more practical support for breast feeding mothers. She would like all new mothers to have the opportunity to stay in a supported setting--not necessarily a hospital--while breast-feeding is established. She recommends finding neutral ways to ensure that all health professionals keep up to date about all aspects of infant nutrition, including infant formulae. She suggests a flexible approach to when weaning starts, since all babies are different and reach developmental milestones at different ages. Advice on weaning is often lacking or conflicting, and needs to be more structured with good quality practical information and leaflets. Finally, the author recommends that babies have meals with the family from an early age, for both social and nutritional reasons. PMID- 16715754 TI - Registered childminders and children with disabilities. AB - This article describes the role of the registered childminder in caring for children with disabilities. It explains the community childminding networks that help put parents in touch with appropriate childminders and then provide continuing support to the parents and the childminder. These networks help provide additional tailored training for childminders to enable them to meet the special needs of a particular child. The networks link in with other local authority and health services and may sometime help to fund placements. PMID- 16715755 TI - [Occupational exposure to hepatitis C virus]. AB - Occupational exposure to Hepatitis C virus is an area of growing concern due to a lack of prophylaxis and limited knowledge regarding prevalence in hospital environment. Between 1999 and 2003, a total of 128 occupational exposures to this virus were registered in Diego Paroissien Hospital, eight of which led to hepatitis (6.3%) and one case to serum conversion (0.8%). Currently there is no preventive therapy and great interest is focused on acute infection therapy the effectiveness of which is still controversial. This study reinforces the fact that adherence to the Universal Precautions is still the most important preventive measure for health care workers, and the most cost beneficial. PMID- 16715756 TI - Periventricular nodular and subcortical neuronal heterotopia in adult epileptic patients. AB - Developmental malformations are brain abnormalities that occur during embryogenesis. Neuronal migration disorders, including heterotopic lesions, constitute one type of such abnormalities. The aim of the study was to compare the epileptic clinical patterns of patients with periventricular nodular heterotopia (PNH) (G1) with those affected by subcortical heterotopia (SCH) (G2) looking for differences between both groups which, eventually, might suggest the type of the underlying malformation. The variables studied in both groups were: type of the heterotopia depicted on MRI studies, sex, age, age at seizure onset, annual seizure frequency, localization of the ictal symptomatogenic zone, characteristics of the EEG, other associated anomalies on the magnetic resonance images (MRI) besides the heterotopia, and response to treatment. The only difference found between both groups was the type of heterotopia as shown by MRI studies. The other assessed variables did not significantly (p>0.05) differ between groups. No differences in the clinical features characterizing epilepsy could be found in patients with PNH or SCH, being the images the only tool able to differentiate them. PMID- 16715757 TI - Regional differences in astrocyte activation in HIV-associated dementia. AB - Since astrogliosis is a histological marker usually observed in HIV-associated dementia (HIV-D), we decided to investigate the potential relationship between the expression of glial fibrillary acidic protein (GFAP) and the regional distribution of cells positive (+) for this specific marker of astrocyte activation. Histological sections of brain tissues obtained at necropsy from 5 HIV-D patients and 5 age-matched controls without history of neuropsychiatric illness were immunostained with peroxidase. Mean numbers of GFAP(+) astrocytes were significantly increased in entorhinal cortex, hippocampus and subcortical white matter of patients, but values in frontal cortex and basal ganglia were similar to those of controls. In contrast, surface density of immunoreactive GFAP was significantly increased in all tested brain areas from all patients, including unusually affected regions such as entorhinal cortex and hippocampus. Therefore, such consistent finding of hypertrophic astrocytes, ranging from highest cell percentajes in subcortical white matter to lowest in basal ganglia indicates that quantification of surface density in GFAP (+) cells appears to be a more reliable approach to score gliosis than the counting of their cell nuclei. Because astrocyte activation involves both protective and detrimental effects on adjacent neuronal subsets, the evidence of regional differences in this reactive potential highlights the importance of accurately defining their contribution to the neuropathogenesis not only of HIV-D, but of a wide range of neurodegenerative disorders. PMID- 16715758 TI - [Gene mixture in a population sample from Buenos Aires City]. AB - The aim of this study is to estimate the gene admixture in the population of Buenos Aires City from samples of blood donors, which come from a public health centre (Hospital de Clinicas). These studies were performed on 218 unrelated people, who donated blood during the year 2002. Eight erythrocyte genetic systems and GM/KM allotypes were analysed. A survey to obtain information about place of birth, present residence and genealogical data of the donors was performed. The gene frequencies were determined using a method of maximum likelihood. The genetic admixture was calculated through the ADMIX program (trihibride). The Amerindian and African contributions were 15.8% and 4.3% respectively. These data were compared with those obtained in a previous study performed in a private centre (Hospital Italiano de Buenos Aires) and significant differences were observed, except in the KM system. The results obtained are in concordance with the demographic and historic information of Buenos Aires City. PMID- 16715759 TI - [Tuberculous meningitis in children under 5 years of age in Argentina]. AB - Tuberculous (TB) meningitis in children under 5 years of age is a serious health problem in Argentina, with a rate of 0.39/100000 inhabitants, for 2003-2004. This rate indicates recent infections. It is an operational indicator for case finding and treatment of pulmonary adult cases, and for BCG vaccination of the newborn. The object of this study was to describe epidemic and clinical variables registered in cases of TB meningitis in children less than 5 years old in Argentina, from 1999 to 2001 and to determine the real number of TB meningitis cases in our country. Fifteen cases of TB meningitis out of 32 (46.8%) were studied through epidemic records. The average age was 14 months. Six children presented BCG scars but in only one child it was proved that he had been vaccinated at birth; 11/13 (92.3%) were at the second stage of illness, the bacteriological investigation was positive in 8/11 (72.7%) and in (46.8%) chest radiography revealed abnormal findings. The tuberculin reaction was negative in all tested cases. This study showed that the diagnosis was made at an advanced stage of disease. None of these patients was cured without sequels and a high death rate (46%) was observed. In order to control this epidemic situation, it is necessary to increase case finding and directly observed treatment of smear positive pulmonary TB cases, as well as BCG vaccination given at birth. PMID- 16715760 TI - Usefulness of recombinant human TSH aided radioiodine doses administered in patients with differentiated thyroid carcinoma. AB - The published studies confirming the safety and efficacy of rhTSH for diagnostic purposes have led to an increased interest in its use for preparation for radioiodine (RI) dose administration in patients with recurrent or persistent differentiated thyroid carcinoma (DTC). In order to establish the efficacy of RI therapy after rhTSH, we have reviewed 39 rhTSH-aided radioiodine treatment in a series of 28 DTC patients. Patients were divided into two groups: GI (n=17), with previous thyroid bed uptake and undetectable thyroglobulin (Tg) levels under levothyroxine treatment and GII (n=11), with proven metastatic local or distant disease. Median follow-up after the first rhTSH-aided radioiodine treatment was 32 +/- 13 months (range 8 to 54 months). Sixteen patients (94%) in GI were rendered disease free and one patient was shown to have persistent disease. In GII, the post therapy whole body scan showed pathological uptakes in all cases: in four patients in lungs, in four in mediastinum and in three in lateral neck. In two patients with mediastinum uptake, Tg levels were undetectable after rhTSH. In the follow-up, two patients with lateral neck uptake were rendered disease free, four patients died (three due to thyroid cancer) and five out of the remaining patients have persistent disease. In conclusion, rhTSH aided therapy was helpful to eliminate normal thyroid bed remnants in 16/17 (94%) patients (GI). rhTSH stimulated Tg was undetectable in two patients with mediastinal metastasis. We believe that rhTSH is a good alternative to levothyroxine withdrawal for the treatment of DTC with radioactive iodine, increasing the quality of life in these patients. Caution should be recommended in the follow-up of unselected DTC patients only with stimulated Tg levels. PMID- 16715761 TI - [Sialidase activity in women with bacterial vaginosis]. AB - Bacterial vaginosis (VB) is a syndrome characterized by overgrowth of endogenous Gram negative bacterial flora and the lack of the normal flora. Within bacterial enzymes, sialidases have been considered a virulence factor of many pathogenic microorganisms colonizing the different mucous membranes. Their presence in vaginal discharges can be correlated with VB. The aim of this study was to detect the activity of this enzyme in women with this syndrome and without clinical evidence of genital infection. Out of a total 112 women studied, 51 were patients with VB and the other 61 women presented normal vaginal flora. For the quantification of enzyme activity, the technique based on the enzymatic hydrolysis of a derivative acid of the acetyl metoxifenil muramic acid was used. In the studied population both groups shared values from 0.5 to 5.1 nmoles of metoxifenol, whereas only 11 out of 52 patients with VB (21.17%), registered more than 5.1 nmoles. The presence of sialidase activity is not enough to confirm VB, except for values greater than 5.5 nmoles of the metoxifenol produced in the enzymatic reaction. PMID- 16715762 TI - [Primary sarcomas of the breast]. AB - Primary sarcomas of the breast are extremely rare with less than 1% of all malignant tumours of the breast reported in literature. At our Institution 1315 malignant tumours of the breast were diagnosed between 1999-2004; nine of them corresponded to primary sarcomas: angiosarcoma (3), leiomyosarcoma (1), low-grade fibromyxoid sarcoma (1), dematofibrosarcoma protuberans (1), liposarcoma (1), osteosarcoma (1), malignant peripheral nerve sheath tumour (1). Histopathological specimens stained with routine techniques and immunoperoxidase were reviewed; proliferation index and p53 over-expression were also determined. Patients' clinical reports were also reviewed to determine prognosis (favorable and unfavorable). The incidence observed (0.7%) is similar to those already published by others authors. Proliferation index was correlated with type of evolution, being an unfavourable prognosis factor when it was equal or major to 30%. Most of the tumours (67%) showed p53 (mayor or equal to 20% of nuclear staining) over expression but this did not show a direct relationship with the evolution of each neoplasm. PMID- 16715763 TI - [Pulmonary mycobacteriosis in non HIV patients, Buenos Aires City, 2003-2004]. AB - Pulmonary disease, due to Mycobacteria other than tuberculosis, is mainly suspected in HIV + patients, or underlying other diseases. In our country, there is no updated information on the prevalence of this pulmonary disease, its treatment and evolution in immucocompetent patients. We present 10 cases of pulmonary disease due to Mycobacteria other than tuberculosis in non HIV patients: clinical-bacteriological diagnosis, treatment and evolution. PMID- 16715764 TI - [Study of the sentinel lymph node. Diagnosis and treatment of cutaneous malignant melanoma stages I and II]. AB - Survival at 5 years of patients with localized primary malignant melanoma is about 80%, compared with a 35% survival in case of lymph nodes involvement. Sentinel lymph node(s) from 45 subjects with diagnosis of malignant melanoma stage I or II was/were studied with hematoxylin-eosin (HE), immunohistochemistry (IHC) and molecular biology (MB) techniques. The population was divided into three groups: HE-/IHC+/MB+, where 67% patients died; HE-/IHC-/MB+, where 57% died; and HE-/IHC-/MB-, where 100% of the patients are alive, with no lymphadenectomy and a median follow-up of 60 months. Those subjects who showed negativity with all the three methods had a null recurrence rate. Data herein obtained suggest a new molecular oncological staging, which would allow the selection of patients with submicroscopic metastases for a complete treatment. Moreover, several patients with no lymph node metastases should not undergo lymphadenectomies, and overtreatment could therefore be avoided. PMID- 16715765 TI - [Pachydermoperiostosis (primary hypertrophic osteoarthropathy)]. AB - Pachydermoperiostosis or primary hypertrophic osteoarthropathy is a rare disease characterized by cutaneous and osteoarthicular involvement. We describe two patients with finger clubbing, watch crystal nails, bones thickenings, arthritis and different grades of skin affection, without other clinical manifestations. Both did not know of having relatives with the same alterations. Radiological studies of the affected areas showed periostosis. Because of normal laboratory results and chest radiography plus the absence of other underlying causes, diagnosis of primary hypertrophic osteoarthropathy was made. PMID- 16715766 TI - [Abdomina distention and edema due to spermatic cord cyst]. AB - Abdominal distention is a frequent symptom, being often the initial presentation of systemic diseases or gastrointestinal disorders. Other causes are uncommon. Spermatic cord cysts are infrequent, abdominal location is even rarer, and the size of the cysts is usually not enough to produce abdominal distention. In our case a man with bilateral cryptorchidism was admitted with abdominal distention and edema of the lower extremities initially interpreted as asciticedematous syndrome. Ultrasonography interpreted abdominal distention as septate ascites, computed tomography as a giant cyst. Exploratory surgery showed a giant spermatic cord cyst in the left spermatic cord. PMID- 16715767 TI - [Bacilar angiomatosis with hepatic peliosis in an AIDS patient]. PMID- 16715768 TI - [Ocular penfigoid scar]. PMID- 16715769 TI - [Response to hypoxia. A systemic mechanism based on the control of gene expression]. AB - New, critically important data have been recently generated about the response to hypoxia. This response can be schematized in three main systems or functions, ie, detectional or oxygen sensing, regulatory, which controls gene expression and effector. The principal organizer of the regulatory branch is a specific transcription factor, the hypoxia-inducible factor 1 (HIF-1). In the presence of oxygen, the alpha subunit of HIF-1 (HIF-1alpha) is modified by hydroxylases, that represent the central point of the oxygen sensing mechanism. This type of hydroxylation induces HIF-1alpha catabolism by the proteosome. On the contrary, in hypoxia, or in the presence of certain growth factors that increase HIF-1alpha synthesis, HIF-1alpha translocates to the nucleus, where it binds HIF-1beta, and thence acts on transcription of genes carrying hypoxia responsive elements (HRE) on their promoters. These genes regulate the synthesis of an ample series of proteins, which span from respiratory enzymes and transporters to hormones regulating circulation and erythropoiesis. The role of HIF-1alpha is not restricted to the mere induction of adaptation to decreased oxygen: instead, it significantly participates in cell repairing mechanisms. A simple list of some of the stimulatory or inhibitory alterations of pathophysiological importance involving the HIF-1 system, would include: chronic lung disease, smoking adaptation, anemia/hemorrhage, ischemia/reperfusion, growth, vascularization and cell resistance of tumors, preeclampsia and intrauterine growth retardation, retinal hyper o hypovascularization, drug intoxications, bowel inflammatory disease and wound repair. This list illustrates by itself the importance of the mechanism herein reviewed. PMID- 16715771 TI - [Nobel Prize of Physiology or Medicine 2005. Acknowledging sharp clinical observation]. PMID- 16715770 TI - [Origin and development of B1 lymphocytes. A cell population involved in defence and autoimmunity]. AB - B1 lymphocytes are an anatomically, phenotypically, and functionally distinct subset of B cells producing the bulk of natural serum IgM in the absence of any apparent stimulation by specific antigens. These cells are a dominant population of B cells in peritoneal and pleural cavities and they have characteristics of activated cells and higher cell size and cytoplasmic complexity than conventional B cells. B1 cells spontaneously secrete antibodies and operate under a differentiation program that is unique and differs from the paradigm associated with Ig-secreting B-2 cells. The antibodies produced by B1 cells may participate in a variety of physiological activities since they are involve in immune regulation, clearance of senescent and apoptotic cells and resistance to infection. However, it has been suggested that they are also involved in autoimmunity. Many advances have been made to describe the origin, development and differentiation of B1 cells, which will be examined here. PMID- 16715773 TI - [Clostridia: not always bad bugs]. PMID- 16715772 TI - [Avian influenza]. PMID- 16715774 TI - [Nobel Prize of Physiology or Medicine 2005]. PMID- 16715775 TI - An improved high-performance liquid chromatographic determination of chloroquine and its major metabolite, desethylchloroquine, in human plasma. AB - Samples were extracted with n-hexane and the organic layer was rejected. 10 microl aliquots of the aqueous layer were injected onto the column. Amodiaquine was used as the internal standard. The UV detector response was linear over the range 0-1000 ng/ml with a correlation coefficient of 0.9987; the detection limits with respect to chloroquine and desethylchloroquine were found to be 20 ng/ml and 10 ng/ml respectively. Within-day and between-day assay variation was generally < 5%. No interference from endogenous constituents was observed. The utility of the method was demonstrated by determining chloroquine and its major metabolite, desethylchloroquine in plasma samples from six healthy human volunteers following a single oral dose of 600 mg of chloroquine. The procedure is simple and requires small volumes of plasma. PMID- 16715776 TI - Absorption enhancement study of astragaloside IV based on its transport mechanism in caco-2 cells. AB - The purpose of this study was to investigate the transport characteristics and mechanisms for discovering the possible causes of the low bioavailability of astragaloside IV and to develop an absorption enhancement strategy. Caco-2 cells used as the in vitro model. Results showed a low permeability coefficient (3.7 x 10(-8)cm/s for transport from the AP to BL direction), which remained unchanged throughout the concentration range studied, indicating that the transport of astragaloside IV was predominantly via a passive route. The AP to BL transport of astragaloside IV was found to be highly sensitive to the extracellular Ca2+ concentration, which suggested that its transport may be via a paracellular route. Both chitosan and sodium deoxycholate can increase the permeation efficiency of astragaloside IV. This study indicated that astragaloside IV having a low fraction dose absorbed in humans mainly due to its poor intestinal permeability, high molecular weight, low lipophilicity as well as its paracelluar transport may directly result in the low permeability through its passive transport. Meanwhile, chitosan and sodium deoxycholate can be used as absorption enhancers based on its transport mechanism. PMID- 16715777 TI - Substrate specificity of guinea pig liver aldehyde oxidase and bovine milk xanthine oxidase for methyl- and nitrobenzaldehydes. AB - Both aldehyde oxidase and xanthine oxidase catalyze the oxidation of a wide range of N-heterocycles and aldehydes. These enzymes are important in the oxidation of N-heterocyclic xenobiotics, whereas their role in the oxidation of xenobiotic aldehydes is usually ignored. The present investigation describes the interaction of methyl- and nitrosubstituted benzaldehydes, in the ortho-, meta- and parapositions, with guinea pig liver aldehyde oxidase and bovine milk xanthine oxidase. The kinetic constants showed that most substituted benzaldehydes are excellent substrates of aldehyde oxidase with lower affinities for xanthine oxidase. Low Km values for aldehyde oxidase were observed with most benzaldehydes tested, with 3-nitrobenzaldehyde having the lowest Km value and 3 methylbenzaldehyde being the best substrate in terms of substrate efficiency (Ks). Additionally, low Km values for xanthine oxidase were found with most benzaldehydes tested. However, all benzaldehydes also had low Vmax values, which made them poor substrates of xanthine oxidase. It is therefore possible that aldehyde oxidase may be critical in the oxidation of xenobiotic and endobiotic derived aldehydes and its role in such reactions should not be ignored. PMID- 16715779 TI - The in vivo metabolism of 3-oxo-5-benzylidene-6-methyl-(4H)-2 (benzoylmethyl)pyridazine in rats. AB - The purpose of the study was to investigate the in vivo metabolic pathway of 3 oxo-5-benzylidene-6-methyl-(4H)-2-(benzoylmethyl)pyridazine (substrate) in rats. Firstly its potential metabolites, i.e. N-dealkylation, ring scission of pyridazine and aromatic hydroxylation products, were synthesized and then the substrate was given orally (100 mg/kg) to male or female Wistar rats at a dose of 100 mg/kg to body weight. Blood samples were collected at 0, 1, 2, 4, 6 and 8 hours after administration of substrate and blood was centrifuged to obtain serum. The substrate and its potential metabolites were separated using a gradient HPLC method on a reverse phase system. This study revealed that 3-oxo-5 benzylidene-6-methyl-(4H)-2-(benzoylmethyl)pyridazine was not metabolized to the proposed metabolites and was present unchanged in the serum. PMID- 16715778 TI - Pharmacokinetics and bioavailability of acyclovir sustained-release tablets in dogs. AB - The pharmacokinetics and bioavailability of acyclovir sustained-release tablets in dogs were investigated. Blood concentrations of acyclovir were determined by RP-HPLC after a single oral dose of two kinds of acyclovir tablets given separately to 6 beagle dogs. The main pharmacokinetics parameters of the acyclovir sustained-release tablet were as follows: T1/2, Tmax and Cmax were 4.10 +/- 0.20 h, 4.05 +/- 0.54 h and 6.90 +/- 0.68 [microg x ml(-1), respectively. MRT was 9.02 +/- 0.44 h. Using acyclovir standard tablet as control, relative bioavailability of the acyclovir sustained-release tablet was 152.2 +/- 49.90%. According to the two-tailed t-test, there was a distinct difference in the data for Tmax, Cmax and AUC between acyclovir sustained-release tablets and acyclovir standard tablets, and the absorbability of acyclovir sustained-release tablets was much better than that of the acyclovir standard tablets. PMID- 16715780 TI - Pharmacokinetic and pharmacodynamic population modeling of orally administered rabeprazole in healthy Chinese volunteers by the NONMEM method. AB - The pharmacokinetic-pharmacodynamic (PK-PD) relationship of the proton pump inhibitor rabeprazole in healthy Chinese volunteers was characterized via a population approach. Healthy Chinese male volunteers were enrolled in the clinical trial. Subjects were divided into three groups by their CYP2C19 genotype. Serum concentrations of rabeprazole were determined using high performance liquid chromatography (HPLC). The intragastric pH values were monitored simultaneously. Data analysis was performed using nonlinear mixed effects modeling as implemented in the NONMEM software package. The final PK-PD model incorporated a one-compartment PK model with one-order absorption from the gastroenteric trace, first-order elimination pathway with one fixed-effect genotype modeling, and a full sigmoidal Emax PD model (X +/- SE: E0 = 2.30 +/- 0.189; Emax = 7.32 +/- 0.662; EC50 = 51.3 +/- 2.142 ng/ml; Hill coefficient = 5.00 +/- 0.556). The time profiles for concentration and pH value, as well as the concentration-pH value relationship of rabeprazole in healthy Chinese volunteers were well described by the developed population PK-PD model. PMID- 16715781 TI - Bioequivalence testing of a new tablet formulation of generic fluoxetine. AB - The pharmacokinetics and relative bioavailability of fluoxetine capsules (reference) and tablets (test) were compared in 24 healthy subjects of both sexes after a single 20 mg oral dose of fluoxetine (as a hydrochloride salt). A randomized, crossover design with a 2-week wash-out period between each dose was applied. Serum samples, obtained before dosing and at various appropriate time points up to 192 hours, were analyzed for fluoxetine and norfluoxetine content by a simple, accurate and precise HPLC method. ANOVA, power analysis, 90% confidence intervals (CI), and two one-sided tests were used for the statistical analysis of pharmacokinetic parameters. The tolerability of the preparations was good. The respective point estimates of the ratios of the geometric means of log-Cmax and log-AUC(0-infinity) of fluoxetine were 0.912 and 0.935 with 90% of 0.838-0.992 and 0.857-1.020. The corresponding point estimates of norfluoxetine were 0.952 (90% CI = 0.843-1.075) and 0.904 (90% CI = 0.807-1.013), respectively. Since both 90% CI for the AUC(0-infinity). and Cmax geometric mean ratios of fluoxetine and norfluoxetine were included in the 80% to 125% interval proposed by the FDA the test drug (fluoxetine tablets) was considered bioequivalent to the reference one (Prozac capsules) according both to the rate and extent of absorption. PMID- 16715783 TI - Promoting development through child health services. Introduction to the Help Me Grow roundtable. PMID- 16715782 TI - Pharmacokinetics/Pharmacodynamics findings after repeated administration of ARTESUNATE thermostable suppositories (RECTOCAPS) in Vietnamese patients with uncomplicated malaria. AB - In recent years, Artemisinin and particularly one of its derivatives--Artesunate (ART--has become an essential alternative for treatment of both uncomplicated and severe falciparum malaria in Asia and Africa as well. Therefore, these compounds are still and inccreasingly in the focus of interest because of quick acting of this drug, is able to help even unconscious to overcome the malaria attack, when administered by injection. As an alternative, RECTOCAPS have been developed and their use is meanwhile well established. From earlier studies in children, suffering from plasmodium falciparum malaria, we obtained a high level of DHART in the blood, but as expected also a rapid decline in the levels of both DHART and ART. A second administration of ART was additionally applied 4 hours after the first administration. DHART and ART plasma levels were found to last longer on an assumed therapeutic level than those obtained after one administration only. The fever clearance and the parasitemia reduction rates were found to be effective according to this dosing regimen. In view of these findings, we decided to conduct the actual described study by administering 200 mg of ART every 3 hours (0, 3, 6 and 9 h) by the rectal route. Soft geiatine capsules (RECTOCAPS) containing 200 mg of ART GMP--type each (Artesunic acid) were administered by rectal route. Each patient received four RECTOCAPS capsules (4 x 200 mg of ART) over a 3 h period. 12 adult patients with uncomplicated malaria were selected. Age, weight, height, body temperature, parasite counts before treatment and their evolution until 96 h are determined. Blood samples were taken at short time intervals after starting with the first medication: 0, 30 min, 60 min, 3 h, 6 h, 9 h, 12 h, 24 h, 36 h, 48 h, 60 h, 72 h, 84 h, 96 h and 108 h. The aliquots of all the blood samples were used for performing parasite counts. Plasma obtained following the traditional procedure was kept at -40 degrees C until analysis. HPLC technique with electrochemical detector was used for quantification of ART and DHART. From the blood concentration values of ART and DHART, the following observation can be derived: the onset of action is observed within the first half hours, therapeutic levels of the drug obtained (89 microg/ml ART compared to 84 microg/ml DHART). The DHART levels are somewhat higher than those of ART (a peak concentration after 6 h starting medication of 151 microg/ml ART as compared to 276 microg/ml DHART). The variations as a function of frequency of DHART uptake are much less marked than those observed for ART. Another finding is that after the administration, some sort of a plateau of DHART and ART is built up, lasting at least from 9 to 12 hours with DHART level of about 190 microg/ml and ART of 90 microg/ml. In the case of single-dose administration, the levels of both compounds were below the detection threshold after three hours. With regard to the parasite counts, although there were inter-individual variations, it should be noted that after 48 hours a high proportion of the patients (8 out of 12) was completely clear of parasites. Similar results were observed with regard to the body temperature (7 out of 12 returned to normal temperature 36 hours after starting the therapy). The findings of the study support the RECTOCAPS application principle resulting in effectiveness both for the velocity of drug uptake as well as for the height of plasma levels. Repeated administration of ART can extend the duration of therapeutic plasma levels of the drug. PMID- 16715784 TI - Historical overview: from ChildServ to Help Me Grow. PMID- 16715786 TI - The role of a statewide information and referral (I&R) system in enhancing the access of children and their families to developmental programs and services. PMID- 16715785 TI - Enhancing developmental services in primary care: the Help Me Grow experience. PMID- 16715787 TI - Promoting change to strengthen developmental outcomes: the role of training. PMID- 16715788 TI - Evaluating model programs to support dissemination. An evaluation of strengthening the developmental surveillance and referral practices of child health providers. PMID- 16715789 TI - Barriers to enhancing practice-based developmental services. PMID- 16715790 TI - Securing and nurturing developmental program and services. PMID- 16715791 TI - Keeping developmental services on the legislative agenda during tight fiscal times. PMID- 16715792 TI - Strengthening health and developmental outcomes in children: the role of philanthropy. PMID- 16715793 TI - Inactivation by ultrahigh-pressure homogenization of Escherichia coli strains inoculated into orange juice. AB - The aim of this work was to evaluate the efficacy of ultrahigh-pressure homogenization (UHPH) for inactivation and/or sublethal injury of two strains of Escherichia coli (O58:H21 ATCC 10536 and O157:H7 CCUG 44857) inoculated into orange juice (pH 3.6). The effects of orange juice inlet temperature (6 and 20 degrees C) on the lethality values and the capacity of these strains for survival, repair, and growth during refrigerated storage after UHPH treatment also was evaluated. Samples of orange juice that had been treated with ultrahigh temperatures were inoculated with E. coli in the stationary phase of growth until a final concentration of approximately 7.0 log CFU/ml was reached. These samples were then treated for one cycle with a double-valve UHPH machine, with 300 MPa at the primary homogenizing valve and 30 MPa at the secondary valve. Counts of viable and injured bacterial cells were obtained for samples taken 2 h after UHPH treatment and after 3, 6, 9, 12, 15, 18, 21, 27, and 33 days of storage at 4 degrees C. The inlet temperature and the strain type both influenced significantly (P < 0.05) the lethality effect on E. coli, which was higher when the inlet temperature was 20 degrees C. No sublethal injuries were detected after any treatment. The changes in viable counts over time for both strains in pressurized and control samples were similar. The viable counts remained high from day 0 to day 18 and then tended to decrease. After 27 days of storage at 4 degrees C, E. coli O157: H7 was more resistant in orange juice samples pressurized at inlet temperatures of 6 and 20 degrees C, with viable counts of 3.41 and 3.20 log CFU/ml, respectively. PMID- 16715794 TI - Survival of a five-strain cocktail of Escherichia coli O157:H7 during the 60-day aging period of cheddar cheese made from unpasteurized milk. AB - The U.S. Food and Drug Administration Standard of Identity for Cheddar cheeses requires pasteurization of the milk, or as an alternative treatment, a minimum 60 day aging at > or =2 degrees C for cheeses made from unpasteurized milk, to reduce the number of viable pathogens that may be present to an acceptable risk. The objective of this study was to investigate the adequacy of the 60-day minimum aging to reduce the numbers of viable pathogens and evaluate milk subpasteurization heat treatment as a process to improve the safety of Cheddar cheeses made from unpasteurized milk. Cheddar cheese was made from unpasteurized milk inoculated with 10(1) to 10(5) CFU/ml of a five-strain cocktail of acid tolerant Escherichia coli O157:H7. Samples were collected during the cheese manufacturing process. After pressing, the cheese blocks were packaged into plastic bags, vacuum sealed, and aged at 7 degrees C. After 1 week, the cheese blocks were cut into smaller-size uniform pieces and then vacuum sealed in clear plastic pouches. Samples were plated and enumerated for E. coli O157:H7. Populations of E. coli O157:H7 increased during the cheese-making operations. Population of E. coli O157:H7 in cheese aged for 60 and 120 days at 7 degrees C decreased less than 1 and 2 log, respectively. These studies confirm previous reports that show 60-day aging is inadequate to eliminate E. coli O157:H7 during cheese ripening. Subpasteurization heat-treatment runs were conducted at 148 degrees F (64.4 degrees C) for 17.5 s on milk inoculated with E. coli O157:H7 at 10(5) CFU/ml. These heat-treatment runs resulted in a 5-log E. coli O157: H7 reduction. PMID- 16715795 TI - Attachment of shiga toxigenic Escherichia coli to beef muscle and adipose tissue. AB - Shiga toxigenic Escherichia coli (STEC) serotypes are important foodborne pathogens that cause gastrointestinal disease worldwide. An understanding of how STEC strains attach to surfaces may provide insight into the potential persistence of and contamination with STEC in food environments. The initial attachment of a selection of STEC serotypes to beef muscle and adipose tissue was evaluated for isolates grown in planktonic and sessile culture. Initial experiments were performed to determine whether attachment differed among STEC strains and between the two modes of growth. Viable counts were obtained for loosely and strongly attached cells, and the strength of attachment (Sr) was calculated. All bacterial isolates grown in sessile culture attached in higher numbers to muscle and adipose tissue than did bacteria in planktonic cultures. For all attachment assays performed, mean concentrations for loosely attached cells were consistently higher than concentrations for strongly attached cells. The mean concentrations for strongly attached bacteria for planktonic and sessile cultures were significantly higher (P < 0.05) on adipose than on muscle tissue. However, some strains of STEC, particularly those from sessile culture, did not differ in their attachment to muscle or adipose tissue. Sr values were not significantly different (P > 0.05) among STEC isolates for all assays. No correlation was found between bacterial hydrophobicity and surface charge values (previously determined) and production of surface structures, viable counts, and Sr values. STEC grown in planktonic and sessile culture seems to behave differently with respect to attachment to muscle and adipose tissue. Cells in sessile culture may have a greater potential to strongly attach to meat surfaces. PMID- 16715796 TI - Evaluation of Escherichia coli O157:H7 growth media for use in test-and-hold procedures for ground beef processing. AB - Since the mid-1990s, the beef industry has used a process called test and hold, wherein beef trim and ground beef are tested to keep products contaminated with Escherichia coli O157:H7 out of commerce. Current O157:H7 detection methods rely on a threshold level of bacterial growth for detection, which is dependent on the growth medium used. Twelve media were examined for growth and doubling time: buffered peptone water (BPW), SOC (which contains tryptone, yeast extract, KCl, MgCl2, and glucose), buffered peptone water plus SOC (BPW-SOC), Bacto-NZYM, RapidChek E. coli O157:H7 medium, BioControl EHEC8 culture medium, Neogen Reveal for E. coli O157:H7--Eight Hour medium (Neogen Reveal 8), BAX System medium for E. coli O157:H7 (BAX) BAX System medium for E. coli O157:H7 MP (BAX-MP), modified E. coli broth, nutrient medium, and tryptic soy broth (TSB). All media were tested at 37 or 42 degrees C under static or shaking conditions. The eight media with the highest total CFU per milliliter and most rapid doubling times were BPW SOC, NZYM, RapidChek, EHEC8, Neogen Reveal 8, BAX, BAX-MP, and TSB. The ability of these eight media to enrich E. coli O157:H7 in ground beef was further evaluated through time-course experiments using immunomagnetic separation. Of these media, TSB was the easiest to prepare, had a wide application base, and was the least expensive. In the test-and-hold process, the normal ratio of medium to product is 1:10. In this study, a 1:3 ratio worked as well as a 1:10 ratio. Processors using test-and-hold procedures could use 1 liter of TSB to enrich for E. coli O157:H7 in a 375-g sample instead of the usual 3.375 liters, thus saving reagents, time, and labor while maintaining accuracy. PMID- 16715797 TI - Multiplication and motility of Salmonella enterica serovars enteritidis, infantis, and montevideo in in vitro contamination models of eggs. AB - The invasive ability of Salmonella enterica serovars Enteritidis, Infantis, and Montevideo in eggs was examined. Strains of these serovars originating from egg contents, laying chicken houses, and human patients were experimentally inoculated (0.1-ml dose containing 78 to 178 cells) onto the vitelline membrane of eggs collected from specific-pathogen-free chickens and incubated at 25 degrees C. The test strains were detected in 25 of 138 yolk contents by day 6, indicating the penetration of Salmonella organisms through the vitelline membrane. There were no significant differences in overall rates of penetration between serovars. The organisms were also detected in the albumen from 125 of 138 eggs tested by day 6. Growth to more than 10(6) CFU/ml was observed in 48 of the 125 albumen samples. An inoculum of 1000 Salmonella cells was added to 15 ml of albumen at the edge of a petri plate. A 10-mm-diameter cylindrical well, the bottom of which was sealed with a polycarbonate membrane with 3.0-microm pores, was filled with egg yolk and placed into the albumen at the center of the dish, which was maintained at 25 degrees C. Experiments were performed in triplicate with each strain. Salmonella organisms in all the albumen samples were detected by day 11. However, motility of the organisms toward the yolk was observed in only two dishes inoculated with the Salmonella Enteritidis strain from a human patient and in one dish inoculated with the Salmonella Infantis strain from liquid egg. The albumen samples obtained from the dishes inoculated with the Salmonella Enteritidis strain had high numbers of bacteria (>10(8) CFU/ml). The present study suggests that Salmonella organisms in egg albumen are unlikely to actively move toward the yolk, although depositionon or near the vitelline membrane can be advantageous for proliferation. PMID- 16715798 TI - Antibiotic resistance profiles and cell surface components of Salmonellae. AB - Salmonellae were isolated from raw chilled retail poultry meats (n=100) using the procedures outlined in the Bacteriological Analytical Manual and Microbiology Laboratory Guidebook. These isolates and 36 Salmonella strains from our laboratory culture collection were tested for their resistance to 12 different antibiotics and for their ability to produce thin aggregative fimbriae and/or cellulose, two of the most important surface components influencing the ability of cells to attach to surfaces and form biofilms. The sensitivity of the salmonellae to the antibiotics was determined with a disc diffusion assay. Of 52 Salmonella isolates, 25 (48.0%) were resistant to one antibiotic, 5 (9.6%) were resistant to two, 4 (7.7%) were resistant to three, 6 (11.5%) were resistant to four, and 5 (9.6%) were resistant to five antibiotics. Two (3.8%) of the isolates were resistant to up to nine of the antibiotics tested. Fifty-one (98%) of the isolates were resistant to novobiocin, 18 (34.6%) were resistant to streptomycin, 14 (26.9%) were resistant to tetracycline, and 14 (26.9%) were resistant to oxytetracycline. In separate experiments, the isolates were grown on Luria Bertani no-salt agar supplemented with Congo red (40 microg/ml) and Coomassie brilliant blue (20 microg/ml) or Calcofluor (200 microg/ml) to determine whether they produced thin aggregative fimbriae and/or cellulose. Of the total 52 Salmonella isolates, 25 expressed only thin aggregative fimbriae, and 1 synthesized only cellulose. Ten isolates produced both thin aggregative fimbriae and cellulose, and the remaining 16 isolates expressed neither surface structure. The findings of this study reveal a prevalence of Salmonella on raw retail poultry products in central Georgia and suggest that salmonellae have the ability to develop resistance to multiple antibiotics and to synthesize cell surface components that help them survive in hostile or suboptimal environments. PMID- 16715799 TI - Effect of direct culture versus selective enrichment on the isolation of thermophilic campylobacter from feces of mature cattle at harvest. AB - Campylobacterjejuni and Campylobacter coli are leading bacterial causes of human gastroenteritis in the United States and other industrialized nations. These organisms frequently colonize avian hosts, including commercial poultry, but are also found in the gastrointestinal tract of other warm-blooded animals, including swine, sheep, and cattle. This study investigated the effect of direct culture versus selective enrichment on the isolation of thermophilic Campylobacter from the colon of 610 cattle. Fecal samples were taken from the colon of mature cattle (older than 30 months of age) immediately after slaughter in a commercial abattoir over a period of 17 months. Campylobacter was isolated from 23.4% of the animals. Most (93%) of the culture-confirmed Campylobacter isolates were C. jejuni, with the remaining 7% being C. coli. Additionally, of the 143 samples from which pure cultures of Campylobacter could be isolated, 72 (50.3%) were positive only with selective enrichment, 18 (12.6%) were positive only with direct plating, and 53 (37.1%) were positive by both methods. The data suggest that, even though selective enrichment was more effective than direct plating, both direct plating and selective enrichment protocols might need to be employed for optimal surveillance of C. jejuni in fecal material from cattle. PMID- 16715800 TI - Genetic diversity of Arcobacter and Campylobacter on broiler carcasses during processing. AB - Broiler carcasses (n=325) were sampled at three sites along the processing line (prescalding, prechilling, and postchilling) in a commercial poultry processing plant during five plant visits from August to October 2004. Pulsed-field gel electrophoresis (PFGE) was used to determine the genomic fingerprints of Camospylobacter coli (n=27), Campylobacter jejuni (n=188), Arcobacter butzleri (n=138), Arcobacter cryaerophilus 1A (n=4), and A. cryaerophilus 1B (n=31) with the restriction enzymes SmaI and KpnI for Campylobacter and Arcobacter, respectively. Campylobacter species were subtyped by the Centers for Disease Control and Prevention PulseNet 24-h standardized protocol for C. jejuni. A modification of this protocol with a different restriction endonuclease (KpnI) and different electrophoresis running conditions produced the best separation of restriction fragment patterns for Arcobacter species. Both unique and common PFGE types of Arcobacter and Campylobacter strains were identified. A total of 32.8% (57 of 174) of the Arcobacter isolates had unique PFGE profiles, whereas only 2.3% (5 of 215) of the Campylobacter isolates belonged to this category. The remaining Arcobacter strains were distributed among 25 common PFGE types; only eight common Campylobacter PFGE types were observed. Cluster analysis showed no associations among the common PFGE types for either genus. Each of the eight common Campylobacter types consisted entirely of isolates from one sampling day, whereas more than half of the common Arcobacter types contained isolates from different sampling days. Our results demonstrate far greater genetic diversity for Arcobacter than for Campylobacter and suggest that the Campylobacter types are specific to individual flocks of birds processed on each sampling day. PMID- 16715801 TI - Counts of Campylobacter spp. on U.S. broiler carcasses. AB - Foodborne Campylobacter-associated gastroenteritis remains a public health concern, and the Centers for Disease Control and Prevention suggests that improperly handled poultry is the most important source of this human disease. In response to these concerns, 10 of the largest U.S. poultry integrators cooperatively determined the incidence and counts of Campylobacter on processed broiler carcasses. Prior to conducting the survey, laboratory personnel were trained in a direct Campy-Cefex plating procedure for enumeration of the organism. Before and after the survey enumeration, consistency in reporting was compared among the participating laboratories. Participating laboratories were able to consistently estimate inoculated concentrations of Campylobacter in carcass rinses. Within the central study, we determined the potential exposure of U.S. consumers to Campylobacter spp. associated with broiler carcasses during a 13-month period. Among each of the 13 participating poultry complexes, rinses from 25 randomly selected fully processed carcasses were sampled monthly from individual flocks. Among 4200 samples, approximately 74% of the carcasses yielded no countable Campylobacter cells. Campylobacter spp. were isolated from approximately 3.6% of all commercially processed broiler carcasses at more than 10(5) CFU per carcass. Acceptable counts of these organisms on raw poultry carcasses remain to be determined. Nevertheless, this survey indicates industry recognition of its responsibility to assess and reduce public exposure to Campylobacter through broiler chickens. PMID- 16715802 TI - Vibrio parahaemolyticus growth under low-iron conditions and survival under high magnesium conditions. AB - Since 1996, Vibrio parahaemolyticus serotype O3:K6 and closely related strains have been associated with an increased incidence of V. parahaemolyticus gastroenteritis worldwide, suggesting the emergence of strains with enhanced abilities to cause disease. One hypothesis for the recent emergence of V. parahaemolyticus O3:K6 and related strains is an enhanced capacity for environmental survival relative to other strains, which might result in increased human exposure to these organisms. Therefore, the objective of this study was to test the hypothesis that survival or growth characteristics of clinical V. parahaemolyticus isolates differ from those of nonclinical isolates under different environmental conditions. Twenty-six V. parahaemolyticus isolates selected to represent either clinical or food sources were monitored for either survival following exposure to high magnesium (300 mM) or growth under iron limited conditions. Isolates in each category (clinical or food) differed widely in survival capabilities following 24 h of exposure to 300 mM Mg2+. Although 4 of 15 clinical isolates grew better at approximately 0.96 microM Fe2+ (iron-limited conditions) than at 50 microM Fe2+ (iron-rich conditions), as an entire group clinical isolates in this study were not more effective at growing under iron limited conditions than were strains not associated with disease. Within the diverse collection of strains examined in these experiments, neither growth characteristics in low-iron environments nor survival capabilities following exposure to high magnesium concentrations were uniformly different between clinical and nonclinical V. parahaemolyticus isolates. Therefore, neither phenotypic characteristic can be used to reliably differentiate potentially pathogenic V. parahaemolyticus strains. PMID- 16715803 TI - Mechanism of action of Spanish oregano, Chinese cinnamon, and savory essential oils against cell membranes and walls of Escherichia coli O157:H7 and Listeria monocytogenes. AB - The mechanism of the antimicrobial action of Spanish oregano (Corydothymus capitatus), Chinese cinnamon (Cinnamomum cassia), and savory (Satureja montana) essential oils against cell membranes and walls of bacteria was studied by the measurement of the intracellular pH and ATP concentration, the release of cell constituents, and the electronic microscopy observations of the cells when these essential oils at their MICs were in contact with Escherichia coli O157:H7 and Listeria monocytogenes. E. coli O157:H7 and L. monocytogenes, two pathogenic foodborne bacteria, were used as gram-negative and gram-positive bacterial models, respectively. Treatment with these essential oils at their MICs affected the membrane integrity of bacteria and induced depletion of the intracellular ATP concentration. Spanish oregano and savory essential oils, however, induced more depletion than Chinese cinnamon oil. An increase of the extracellular ATP concentration was observed only when Spanish oregano and savory oils were in contact with E. coli O157:H7 and L. monocytogenes. Also, a significantly higher (P < or = 0.05) cell constituent release was observed in the supernatant when E. coli O157:H7 and L. monocytogenes cells were treated with Chinese cinnamon and Spanish oregano oils. Chinese cinnamon oil was more effective to reduce significantly the intracellular pH of E. coli O157:H7, whereas Chinese cinnamon and Spanish oregano decreased more significantly the intracellular pH of L. monocytogenes. Electronic microscopy observations revealed that the cell membrane of both treated bacteria was significantly damaged. These results suggest that the cytoplasmic membrane is involved in the toxic action of essential oils. PMID- 16715804 TI - Combined treatment of high pressure and heat on killing spores of Alicyclobacillus acidoterrestris in apple juice concentrate. AB - Alicyclobacillus acidoterrestris, a thermoacidophilic and spore-forming bacterium, has been isolated from spoiled acidic juices and is considered to be one of the important target microorganisms in quality control of acidic canned foods. Combined high pressure and heat treatment showed an effectiveness to control A. acidoterrestris spores. However, the effectiveness of the combined treatment may change upon the apple juice concentration. Therefore, the objective of this study was to evaluate different levels of apple juice concentrate for reduction of Alicyclobacillus spores by high pressure and heat. Spores of A. acidoterrestris were inoculated into three different concentrations of apple juice (17.5, 35, and 70 degrees Brix), and subjected to three high-pressure treatments (207, 414, and 621 MPa) at four different temperatures (22, 45, 71, and 90 degrees C). High-pressure treatment (207, 414, and 621 MPa) at 22degrees C did not reduce the level of spores regardless of the apple juice concentration (P > 0.05). In diluted apple juice (17.5 degrees Brix), the combined treatment of high pressure and heat resulted in spore reductions of about 2 log at 45 degrees C, and more than 5 log at higher temperatures (71 and 90 degrees C) in a high pressure and temperature-dependent manner. For apple juice with a higher concentration (30 degrees Brix), high-pressure treatment showed no effect at 45 degrees C but resulted in about 2 and 4 log reduction at 71 and 90 degrees C, respectively. However, for apple juice concentrate (70 degrees Brix), treatment with heat or high pressure alone, or their combinations showed no inactivation against spores of A. acidoterrestris. It is likely that differences in the water availability explain the greater resistance of spores to high-pressure inactivation in the juice concentrates than in diluted juices. Our results demonstrate that the effect of high pressure combined with heat against spores of A. acidoterrestris was highly dependent on the apple juice concentration. PMID- 16715805 TI - Inhibitory effect of select nitrocompounds on growth and survivability of Listeria monocytogenes in vitro. AB - We report the effects of 2-nitro-1-propanol (2NPOH), 2-nitroethanol (2NEOH), and nitroethane (NE) on growth and survivability of Listeria monocytogenes. In all cases, inhibition was greatest with 2NPOH and least with NE. For example, specific growth rates of L. monocytogenes strain 18 declined (P < 0.05) 76, 60, and 29% from controls during aerobic culture at 37 degrees C in brain heart infusion broth containing 10 mM 2NPOH, 2NEOH, or NE, respectively. Mean specific growth rate for the controls incubated likewise without added nitrocompound was 0.62 +/- 0.02 h(-1). Specific growth rates of L. monocytogenes Scott A decreased (P < 0.05) 67, 45, and 11%, respectively, from controls (0.67 +/- 0.02 h(-1)) when cultured similarly. Specific growth rates for L. monocytogenes strain 18 incubated similarly except at 30 degrees C were reduced (P < 0.05) 76, 60, and 30%, respectively, and were reduced (P < 0.05) 78, 23, and 23% during anaerobic culture at 30 degrees C in brain heart infusion broth containing 15 mM 2NPOH, 2NEOH, or NE (control rates ranged from 0.37 +/- 0.07 to 0.74 +/- 0.05 h(-1)). Survivability of L. monocytogenes strain 18 was reduced (P < 0.05) during aerobic storage (4 months at 4 degrees C) in brain heart infusion broth containing 2NPOH or 2NEOH (by 7.8 and 1.9 log units, respectively) but not NE. The inhibitory effect of 2NPOH was approximately 20% greater during growth at pH 7.0 than at pH 5.6 or 8.0. These results demonstrate the differential inhibitory activity of 2NPOH, 2NEOH, and NE against L. monocytogenes in vitro. PMID- 16715806 TI - Bacteriocin activity by Lactobacillus curvatus CWBI-B28 to inactivate Listeria monocytogenes in cold-smoked salmon during 4 degrees C storage. AB - The inhibition effectiveness of a bacteriocin produced by Lactobacillus curvatus CWBI-B28 against Listeria monocytogenes was investigated in cold-smoked salmon during storage at 4 degrees C. Three bacteriocin-based strategies for the control of L. monocytogenes in foods (i.e., producing bacteriocin in situ, spraying with partially purified bacteriocin, and packaging in bacteriocin-coated plastic film), plus a newly developed method that uses cell-adsorbed bacteriocin (i.e., a suspension of producer cells on which maximum bacteriocin has been immobilized by pH adjustments), were assessed. Although all the approaches inactivated L. monocytogenes in cold-smoked salmon, various efficacy levels were observed. The behavior of L. monocytogenes was similar in samples treated with either partially purified bacteriocin or in situ bacteriocin production. In both of these cases, the counts of the pathogen declined to below the detectable limit of 0.7 log CFU/cm2 within the first week, but a approximately 0.95- and 1.3-log increase, respectively, occurred after day 14. The bioactive packaging film resulted in a slower inactivation of the pathogen but prevented any subsequent increase in the CFU throughout 22 days of storage at 4 degrees C. Application of the cell adsorbed bacteriocin was shown to be the most effective means, as it resulted in a complete inactivation of the pathogen within 3 days, and no increase in Listeria counts occurred up to 22 days. PMID- 16715807 TI - PCR primers for the detection of staphylococcal enterotoxins K, L, and M and survey of staphylococcal enterotoxin types in Staphylococcus aureus isolates from food poisoning cases in Taiwan. AB - Staphylococcal enterotoxins (SEs) are important causative agents in gastroenteritidis and food poisoning cases. They are serologically grouped into five major classical types, i.e., SEA, SEB, SEC, SED, and SEE. In addition, new SEs, such as SEG through SEM, have recently been identified and characterized. In an attempt to survey the distribution of classical and new SEs in organisms responsible for staphylococcal infections in Taiwan, we developed PCR primers for the genes that define the SEK, SEL, and SEM types. Bacterial strains other than sek, sel, and sem Staphylococcus aureus, including strains of other Staphylococcus species, did not generate any false-positive results when examined with these primers. The expression potential for the sek, sel, and sem types were also determined by reverse transcription-PCR. Together with the PCR primers specific for the classical SEs and other new SEs, including SEG, SEH, SEI, and SEJ, we surveyed the SE genes in S. aureus strains isolated from food poisoning cases. For 147 S. aureus isolates originating from food poisoning cases, 109 (74.1%) were positive for one or more SE genes. Of them, the major classical enterotoxin type was sea (28.6%), followed by seb (20.4%), sec (8.2%), and sed (2.0%). For the new SE types, sei (30.6%) was detected the most often, followed by sek (18.4%), sem (12.9%), and sel (8.2%). Also, 64 (43.5%) of the total bacterial strains had more than one enterotoxin gene. PMID- 16715808 TI - Thermal inactivation studies of Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes in ready-to-eat chicken-fried beef patties. AB - Thermal inactivation studies were used to determine the D- and z-values of Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes in ready-to-eat chicken-fried beef patties. Inoculated meat was packaged in sterile bags, which were immersed in a circulated water bath and held at 55, 57.5, 60, 62.5, 65, 67.5, and 70 degrees C for different lengths of time. D- and z-values were determined with a linear regression model. Average D-values at temperatures 55 to 70 degrees C were 27.62 to 0.04 min for E. coli 0157:H7, 67.68 to 0.22 min for Salmonella, and 81.37 to 0.31 min for L. monocytogenes. The z-values were 5.2 degrees C for E. coli O157:H7, 6.0 degrees C for Salmonella, and 6.1 degrees C for L. monocytogenes. The results of this study can be used by food processors to validate their processes and help eliminate pathogenic bacteria associated with chicken-fried beef products. PMID- 16715809 TI - Pathogen survival in chorizos: ecological factors. AB - This study addressed health risks from ethnic sausages produced on a small scale, without inspection, in California and elsewhere. Mexican-style chorizo, a raw pork sausage that is not cured, fermented, or smoked, was contaminated experimentally in the batter with Escherichia coli O157:H7, Listeria monocytogenes, or Salmonella serotypes and stuffed into natural casings. Formulations were based on a market survey in California. Physical parameters that were controlled were pH, water activity (a(w)), and storage temperature. The pH was adjusted with vinegar, stabilizing at 5.0 within 24 h. Initial a(w) levels adjusted with salt were 0.97, 0.95, 0.93, 0.90, and 0.85; levels declined with time because of evaporation. Pathogen numbers declined with storage up to 7 days, with few brief exceptions. Main effects and interactions of constant temperature and pH with declining a(w) on survival of the pathogens were determined. Maximum death rates occurred at higher a(w) for E. coli O157:H7 and Salmonella than for L. monocytogenes. Salt used to adjust a(w) affected palatability. Spices (black pepper, chili pepper, chili powder, cumin, garlic, guajillo pepper, oregano, and paprika) comprised another, potentially significant aspect of the sausage formulation. Some (notably black pepper and cumin) carried an indigenous microflora that contributed significantly to the microbial load of the sausage batter. Only undiluted fresh and powdered garlic exhibited a significant antimicrobial effect on the pathogens. Although each of the tested formulations caused death of the inoculated pathogens, none of the death rates was sufficiently rapid to ensure safety within the probable shelf life of the product. PMID- 16715810 TI - Occurrence of selected foodborne pathogens on poultry and poultry giblets from small retail processing operations in Trinidad. AB - We conducted a study to determine quantitatively and qualitatively the presence of Campylobacter spp., Escherichia coli, staphylococci, total coliforms, total aerobic bacteria, and Salmonella on broiler carcasses from selected small retail processors in Trinidad. We used standard media and procedures for detection and quantification. All carcass and weep samples were positive for aerobic bacteria, E. coli, total coliforms, and staphylococci. Significant differences in the mean counts of aerobic bacteria were observed for samples of carcass (P = 0.001), weep (P = 0.038), and liver and heart (P = 0.017). There was a significant difference (P < 0.05) in the prevalence of E. coli and Campylobacter for liver and heart samples and gizzard samples across various areas (health divisions) in Trinidad and for Campylobacter jejuni and Campylobacter coli for offal samples. The prevalence of Salmonella in carcass, drip, gizzard, and liver and heart samples was 7.3, 3.1, 2.1, and 1.0%, respectively, and three serotypes, Salmonella Kiambu (53.8%), Salmonella Kentucky (38.5%), and Salmonella Mbandaka (7.7%) were isolated. Of the six groups of microbes considered with respect to sale activity, the differences in the prevalence of Campylobacter in medium-activity sale shops (95.8%) and low-activity sale shops (83.3%) and the mean counts of staphylococci for medium-activity sale shops (5.5 +/- 0.9) and low-activity sale shops (5.1 +/- 0.8) were statistically significant (P < 0.05). Carcasses rinsed in a stagnant system had a significantly higher (P < 0.05) prevalence (92.3%) and mean count per milliliter (3.1 +/- 0.7) for Campylobacter compared with 77.8% and 2.7 +/- 0.7 for shops that rinsed with constantly running water. The frequency of rinse water change significantly (P = 0.04) affected the prevalence of Salmonella on carcasses. It is recommended that a quality control system be introduced for these shops, particularly with respect to evisceration and rinsing practices. PMID- 16715811 TI - Rabbit meat as a source of bacterial foodborne pathogens. AB - Even though worldwide production of rabbit meat is >1,000,000 tons, little information is available for rabbit meat microbiology. This study provides data on the prevalence of Salmonella, Escherichia coli O157:H7, Yersinia enterocolitica, Listeria spp., motile Aeromonas spp., and Staphylococcus aureus on rabbit meat. A total of 24 rabbit carcasses from two abattoirs and 27 rabbit meat packages from supermarket displays were examined. In addition to culturing methods, associated virulence genes were investigated by PCR in suspect isolates and samples. Neither Salmonella nor E. coli O157:H7 was detected. All samples were negative for virulence-associated invA, stx1, and stx2 genes. At one abattoir, two carcasses (3.9%) carried Y. enterocolitica yst-, and two were positive for the yst gene, although viable Y. enterocolitica cells were not recovered from these samples. Seven samples (13.7%) were contaminated with Listeria. Of them, three were positive for hly and iap genes (Listeria monocytogenes hly+ / iap+), two carried Listeria seeligeri, one carried Listeria ivanovii, and one carried Listeria innocua. For detectable motile Aeromonas spp. (average count, 1.77 +/- 0.62 log CFU/g), the contamination rate was 35.3%, although ca. 90% of the samples were positive for the aerA and/or hlyA genes. The majority of aeromonad isolates were Aeromonas hydrophila aerA+ / hlyA+. Aeromonas caviae, Aeromonas popoffii, Aeromonas schubertii, and the two biovars of Aeromonas veronii were also isolated. The prevalence of S. aureus contamination (average count, 1.37 +/- 0.79 log CFU/g) was 52.9%. Among 27 S. aureus isolates, two harbored genes for staphylococcal enterotoxin B (seb), and two harbored genes for staphylococcal enterotoxin C (sec). The remaining isolates were negative for sea, seb, sec, sed, and see. PMID- 16715812 TI - A national survey of the microbiological quality of beef carcasses and frozen boneless beef in Australia. AB - The third national baseline microbiological survey of Australian beef carcasses and frozen boneless beef was conducted in 2004. Carcasses (n=1155) sampled at 27 slaughter establishments had a mean aerobic plate count (at 25 degrees C) of 1.3 log CFU/cm2. Escherichia coli was isolated from 8.0% of the cacasses, with a mean count of -0.8 log CFU/cm2 for positive samples. On samples from 24 boning (fabrication) plants (n=1082), the mean aerobic plate count for frozen boneless beef was 1.3 log CFU/g, and the mean count for the 1.8% of samples with detectable E. coli was 1.5 log CFU/g. E. coli O157: H7 was isolated from 1 of 1,143 carcasses and from 0 of 1082 boneless samples. Salmonella was isolated from 0 of 1155 carcasses and from 1 of 1082 samples of boneless product. No Campylobacter spp. were isolated from carcasses or boneless beef. Coagulase positive staphylococci were isolated from 28.7% of beef carcasses and 20.3% of boneless beef samples, and positive samples had a mean count of 0.3 log CFU/cm2 and 0.8 log CFU/g, respectively. PMID- 16715813 TI - Monitoring the microbial populations and temperatures of fresh broccoli from harvest to retail display. AB - Microbial populations and the temperature of fresh broccoli were monitored at several steps of a supply chain by sampling 33 distinct lots of locally grown produce over two seasons during harvest, storage, wholesale handling, and retail display. Imported broccoli was also sampled, but only at retail display. Microbiological analyses were conducted on the florets of 201 local and 60 imported broccoli samples to determine populations of total aerobic bacteria (aerobic colony count), fecal coliforms, Escherichia coli, and Listeria monocytogenes. All the samples had mean aerobic colony counts ranging between 4 and 6 log CFU/g, but L. monocytogenes was not detected (limit of detection =100 CFU/g). Fecal coliforms and E. coli (limit of detection =20 most probable number per 100 g) were found in 22 of 126 samples of local broccoli collected at various steps of the production and distribution system during the first season. None was found in 75 samples collected in the second season. Fecal coliforms and E. coli were found in 2 of 60 imported broccoli samples. Broccoli temperatures were relatively well controlled throughout the production and distribution system. No clear change in produce microbial populations was evident between harvest and retail display, during both sampling seasons. However, a large experimental variability was found, possibly associated with the high variability of the initial levels of microbial populations on broccoli at harvest. PMID- 16715814 TI - Effect of low-dose radiation on microbiological, chemical, and sensory characteristics of chicken meat stored aerobically at 4 degrees C. AB - The effect of gamma-radiation (0.5, 1, and 2 kGy) on the shelf life of fresh skinless chicken breast fillets stored aerobically at 4 degrees C was evaluated. Microbiological, chemical, and sensorial changes occurring in chicken samples were monitored for 21 days. Irradiation reduced populations of bacteria, i.e., total viable bacteria, Brochothrix thermosphacta, lactic acid bacteria (LAB), and the effect was more pronounced at the highest dose (2 kGy). Pseudomonads, yeasts and molds, and Enterobacteriaceae were highly sensitive to gamma-radiation and were completely eliminated at all doses. Of the chemical indicators of spoilage, thiobarbituric values for nonirradiated and irradiated aerobically packaged chicken samples were in general low (<1 mg of malonaldehyde per kg of muscle) during refrigerated storage for 21 days. With regard to volatile amines, both trimethylamine nitrogen (TMA-N) and total volatile basic nitrogen (TVB-N) values for nonirradiated aerobically packaged chicken increased steeply, with final values of ca. 20.3 and 58.5 mg N/100 g of muscle, respectively. Irradiated aerobically packaged chicken samples had significantly lower TMA-N and TVB-N values (P < 0.05) of ca. 2.2 to 3.6 and 30.5 to 37.1 mg N/100 g of muscle, respectively, during refrigerated storage for 21 days. Of the biogenic amines monitored, only putrescine and cadaverine were detected in significant concentrations in both nonirradiated and irradiated chicken samples, whereas histamine formation was noted only in nonirradiated samples throughout storage. On the basis of sensorial evaluation, low-dose irradiation (0.5 and 1.0 kGy) in combination with aerobic packaging extended the shelf life of fresh chicken fillets by ca. 4 to 5 days, whereas irradiation at 2.0 kGy extended the shelf life by more than 15 days compared with that of nonirradiated chicken. PMID- 16715815 TI - Polycyclic aromatic hydrocarbons in fresh and cold-smoked Atlantic salmon fillets. AB - The occurrence of polycyclic aromatic hydrocarbons (PAHs) in smoked fish as a consequence of cold smoking was studied. Raw fillets of Salmo salar from Norway or the Irish Sea were sampled in a modern smokehouse and examined for PAH content. The same fillets, labeled with an identification number, were sampled immediately after the smoking process and analyzed. Among the investigated compounds, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, benz[a]anthracene, chrysene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene, and benzo[ghi]perylene were detected in both raw and smoked fillets. No significant difference (P < 0.01) was observed between raw and smoked samples in the concentrations of six PAHs, but significant differences were found for fluorene, anthracene, fluoranthene, benz[a]anthracene, and benzo[ghi]perylene. Results confirm that PAHs concentrations in smoked fish are the product of both sea pollution and the smoking process. A modern smoking plant with an external smoke generator and a mild treatment as described here will not add significantly to the concentration of PAHs, except for some compounds. PMID- 16715816 TI - Reduction of ochratoxin A in extruded barley meal. AB - The objective of this work was to determine the effects of extrusion cooking on the stability of ochratoxin A (OTA) in an artificially contaminated hulled barley meal (0.73-mm grain diameter) using a single screw extruder. The extrusion cooking parameters were temperature (140, 160, and 180 degrees C), initial moisture content of barley meal (24, 27 and 30%), and residence time (30, 40, 50, 60, and 70 s). Both unextruded and extruded samples were analyzed for OTA by high performance liquid chromatography. Extrusion cooking variables significantly affected the stability of OTA (P < 0.05). Greater OTA reductions were achieved at higher residence time (70 s), medium temperature level (160 degrees C), and either high (30%) or low moisture (24%) content of samples. The amount of OTA destroyed during the extrusion process ranged from 17 to 86% depending on the studied parameters. The decrease in the amount of OTA after extrusion cooking followed first-order kinetics, showing that the fastest treatment in OTA reduction was that at 140 degrees C and 24% of moisture content. PMID- 16715817 TI - Assessment of edible marine species in the Adriatic Sea for contamination from polychlorinated biphenyls and organochlorine insecticides. AB - It is estimated that 90% of human exposure to persistent organic pollutants is through food, and fish and shellfish represent an important source of contamination for polychlorinated biphenyls (PCBs) and organochlorine insecticides. To evaluate the levels of seafood contamination coming from the central Adriatic Sea, Italy, a study involving several pools of shellfish, crustaceans, and fish was carried out. Several marine species were selected by their abundance, wide distribution, and common use in the Italian diet and sampled and analyzed during 2004. The concentration of total (sigma) PCBs exceeded that of total dichlorodiphenyltrichloroethanes (DDTs) in all samples. Atlantic mackerel showed the highest concentrations of PCBs, ranging from 514 to 1772 ng/g of fat weight, and DDTs, ranging from 52 to 656 ng/g of fat weight. The lowest concentrations of PCBs and DDTs were found in cephalopods and mussels. Despite this, to protect human health from these pollutants, legal limits have been established for fish and shellfish for DDTs but not PCBs. The most common representative PCB congeners, in all species, were PCB 153 and PCB 138; the most common DDT was p,p'-dichlorodiphenyldichloroethylene. PMID- 16715818 TI - A summary of national reports of foodborne outbreaks of Salmonella Heidelberg infections in the United States: clues for disease prevention. AB - We analyzed national foodborne outbreak data from 1973 through 2001 to determine the proportion of Salmonella Heidelberg outbreaks caused by specific foods. Among 6633 outbreaks with known etiology, 184 (3%) were caused by Salmonella Heidelberg. A vehicle was identified in 101 outbreaks; at least 53 were poultry or egg-related. Three outbreaks were attributed to egg consumption, 17 to consumption of egg-containing foods, 25 to poultry, and 8 to foods containing poultry and eggs. Efforts to reduce illness due to Salmonella Heidelberg shouldensure that poultry and eggs are handled appropriately to minimize contamination and cross contamination. PMID- 16715819 TI - Fecal shedding of Escherichia coli O157:H7 in North Dakota feedlot cattle in the fall and spring. AB - Cattle are an important reservoir of Escherichia coli O157:H7, which can lead to contamination of food and water, and subsequent human disease. E. coli O157:H7 shedding in cattle has been reported as seasonal, with more animals shedding during summer and early fall than during winter. North Dakota has relatively cold weather, especially in winter and early spring, compared with many other regions of the United States. The objective was to assess fecal shedding of E. coli O157:H7 in North Dakota feedlot cattle over the fall, winter, and early spring. One hundred forty-four steers were assigned randomly to 24 pens on arrival at the feedlot. Samples of rectal feces were obtained from each steer four times (October and November 2003, and March and April 2004) during finishing. On arrival (October 2003), 2 (1.4%) of 144 cattle were shedding E. coli O157:H7. The shedding increased significantly to 10 (6.9%) of 144 after 28 days (November 2003), to 76 (53%) of 143 at the third sampling (March 2004), and dropped significantly to 30 (21%) of 143 at the fourth (last) sampling (March 2004) before slaughter. Unfortunately, we were unable to sample the cattle during winter because of the extreme weather conditions. Sampling time significantly (P < 0.0001) influenced variability in E. coli O157:H7 shedding, whereas herd (P = 0.08) did not. The prevalence of E. coli O157:H7 shedding in North Dakota steers in fall and early spring was comparable to what has been reported in other parts of the United States with relatively warmer weather. Further research into E. coli O157:H7 shedding patterns during extreme weather such as North Dakota winters is warranted in order to fully assess the seasonal effect on the risk level of this organism. PMID- 16715820 TI - Survival of Streptococcus pyogenes on foods and food contact surfaces. AB - Streptococcus pyogenes causes septic sore throat in millions of Americans each year and may be transmitted from food handlers to food contact surfaces, foods, and consumers. This study examined the individual survival of six S. pyogenes strains on food contact surfaces (plastic and ceramic plates, plastic cups, and stainless steel utensils) held at 21 degrees C for 2 h and on tomatoes stored aerobically at 21 degrees C for 2 h and at 5 degrees C for 24 h. Survival of a cocktail of the six S. pyogenes strains was also evaluated on vacuum-packaged ready-to-eat meats and cheeses held at 21 degrees C for 8 h and at 5 degrees C for 24 h. Populations generally did not change on tomatoes, cheeses, or beef bologna; however, there were small (0.1 to 0.7 log CFU) but statistically significant decreases (P < 0.05) in average S. pyogenes populations on turkey luncheon meat and beef summer sausage stored for 8 h at 21degrees C and on beef summer sausage stored for 24 h at 5 degrees C. On food contact surfaces, average populations either decreased slightly (P > or = 0.05) or remained constant, with the exception of three strains that significantly decreased in number on ceramic plates (P < 0.05; average decreases, 0.3 log CFU). Results of this study suggest the importance of preventing the contamination of foods and food contact surfaces with S. pyogenes by infected workers. PMID- 16715821 TI - Persistence of Clostridium botulinum neurotoxin type E in tissues from selected freshwater fish species: implications to public health. AB - Rainbow trout (Oncorhynchus mykiss), round gobies (Neogobius melanostomas), yellow walleye (Stizostedion vitreum), and yellow perch (Perca flavescens) were given Clostridium botulinum neurotoxin type E (BoNT/E) at four doses (0, 800, 1500, and 4000 mouse lethal doses). BoNT/E was sought in the fish tissues at death or at the conclusion of the experiment (10 days after treatment). Fish were divided into a "fillet" (axial musculature) and a "nonfillet" sample before testing for BoNT/E toxicity with a mouse bioassay. BoNT/E was detected in all species. The percentage of positive BoNT samples ranged across the species and doses from 0 (trout, perch, and walleye) to 17% (round goby) in fillet tissues and from 0 (perch) to 92% (round goby) in nonfillet tissues. The lack of positive fillet samples in three key commercial fish species suggests that the public health implications of eating these fish are minimal. However, the presence of toxin in the nonfillet compartment of a high proportion of fish supports the hypothesis that live intoxicated fish are a vehicle for the transfer of BoNT/E to fish-eating birds, which are then in turn, intoxicated. PMID- 16715822 TI - Biofilm-spore response in Bacillus cereus and Bacillus subtilis during nutrient limitation. AB - This study aimed to trace the dynamics of biofilm formation by vegetative cells and endospores of Bacillus cereus DL5 and Bacillus subtilis 168. Counts of B. cereus DL5 and B. subtilis 168 vegetative cells and spores either attached to glass wool or, correspondingly, planktonic cells were determined by standard plate-counting methods. Results from this study highlighted the biofilm-forming potential of both spores and vegetative cells of two different Bacillus species. It was shown that once Bacillus spores had attached to a surface, the spores germinated under favorable (B. cereus DL5) and even unfavorable (B. subtilis 168) nutrient conditions, resulting in biofilms containing both spores and vegetative populations. Furthermore, it was suggested that vegetative B. cereus DL5 cells exhibited a low propensity for spore formation in attached and planktonic growth forms in nutrient-limited growth medium. By contrast, vegetative B. subtilis 168 cells readily formed spores in planktonic and attached microcosms when exposed to nutrient-limited growth conditions. Sporulation in attached Bacillus populations is an important practical consideration for many food industries, such as dairy processing, where bacilli are routinely isolated from populations attached to processing-equipment surfaces. PMID- 16715823 TI - Presence and growth of Bacillus cereus in dehydrated potato flakes and hot-held, ready-to-eat potato products purchased in New Zealand. AB - Potato products prepared from dehydrated potato flakes have been implicated in foodborne illness incidents involving Bacillus cereus intoxications. B. cereus can survive as spores in potato flakes and can germinate and multiply in the rehydrated product. This study assessed the frequency and concentration of B. cereus in dehydrated potato flakes and hot-held, ready-to-eat mashed potato products. Of 50 packets of potato flakes tested, eight contained greater than 100 CFU/g B. cereus (maximum 370 CFU/g). The temperature of the potato portion of 44 hot-held food products was measured immediately after purchase, and 86% were below the safe hot-holding temperature of 60 degrees C. The potato portions were subsequently tested for B. cereus. Only two of the potato portions contained B. cereus at greater than 100 CFU/g, a potato-topped pastry (1000 CFU/g) and a container of potato and gravy (120 CFU/g). To assess multiplication of B. cereus in this food, we held rehydrated potato flakes with naturally occurring B. cereus at 37, 42, and 50 degrees C and tested them over 6 h. By 6 h, the number of B. cereus in potato stored at 37 degrees C had exceeded 10(3) CFU/g, was greater than 10(4) CFU/g at 50 degrees C, and was close to 10(6) CFU/g at 42 degrees C. Growth data were compared to predictions from the U.S. Department of Agriculture Pathogen Modeling Program (PMP 7.0). The PMP predictions were found to simulate the measured growth better at 42 degrees C than at 37 degrees C. Hot-held potato products should be safe for consumption if held at 60 degrees C or above or discarded within 2 h. PMID- 16715824 TI - Plasmid profile patterns and properties of pediococci isolated from caper fermentations. AB - A collection comprising 14 isolates of Pediococcus pentosaceus and one Pediococcus acidilactici from the fermentation of caper fruits was studied. All isolates showed very similar fermentation profiles and produced a limited number of exoenzymes. All isolates carried large plasmids of diverse sizes between 20 and 55 kb, while some also contained smaller plasmids between 10 and 16 kb. Cluster analysis of plasmid profiles revealed four main groups with various degrees of similarities. All amino acid decarboxylation tests were negative, suggesting that pediococci are not involved in generation of biogenic amines. None of the isolates showed hemolytic activity. Antimicrobial resistance tests revealed that all isolates were sensitive to 11 different antimicrobials while being resistant to ciprofloxacin (MIC > or =2 mg/liter) and intrinsically resistant to vancomycin (MIC > or =16 mg/liter) and teicoplanin (MIC > or =16 mg/liter). PMID- 16715825 TI - Microbiological profiles, pH, and titratable acidity of chorizo and salchichon (two Spanish dry fermented sausages) manufactured with ostrich, deer, or pork meat. AB - Microbial counts, pH, and titratable acidity were determined in 102 Spanish dry fermented sausages (chorizo and salchichon) made with ostrich, deer, or pork meat. Average microbial counts (log CFU per gram) varied from 5.46 +/- 0.24 to 8.25 +/- 0.80 (total viable counts), from 4.79 +/- 0.36 to 7.99 +/- 0.20 (psychrotrophs), from 0.00 +/- 0.00 to 0.99 +/- 1.10 (undetectable values were assumed to be zero) (Enterobacteriaceae), from 0.00 +/- 0.00 to 4.27 +/- 1.47 (enterococci), from 5.15 +/- 1.15 to 8.46 +/- 0.49 (lactic acid bacteria), from 3.08 +/- 0.44 to 6.59 +/- 1.76 (Micrococcaceae), from 2.27 +/- 1.53 to 5.11 +/- 1.81 (molds and yeasts), from 0.00 +/- 0.00 to 2.25 +/- 0.81 (pseudomonads), and from 0.00 +/- 0.00 to 2.78 +/- 0.46 (Brochothrix thermosphacta). Average pH and titratable acidity varied from 5.07 +/- 0.25 to 5.63 +/- 0.51 (pH units) and from 0.30 +/- 0.01 to 0.86 +/- 0.19 (% lactic acid). Both type of sausage (P < 0.05) and species of meat (P < 0.001) influenced microbial counts. Salchich6n samples showed lower average values than chorizo samples for most microbial groups (significant for Enterobacteriaceae, lactic acid bacteria, and B. thermosphacta) and titratable acidity. Sausages made from pork showed the highest microbial loads for total viable counts, psychrotrophs, Enterobacteriaceae, enterococci, lactic acid bacteria, and yeasts and molds. Higher counts were observed only for pseudomonads in ostrich sausages. B. thermosphacta levels were similar for all species of meat. The highest average pH value was observed in sausages made from ostrich meat, and the lowest titratable acidity level was found in pork sausages. PMID- 16715826 TI - Requisite scientific parameters for establishing the equivalence of alternative methods of pasteurization. PMID- 16715827 TI - Fear: a factor to consider in musculoskeletal rehabilitation. PMID- 16715828 TI - Clinical diagnosis of an anterior cruciate ligament rupture: a meta-analysis. AB - STUDY DESIGN: Meta-analysis. OBJECTIVES: To define the accuracy of clinical tests for assessing anterior cruciate ligament (ACL) ruptures. BACKGROUND: The cruciate ligaments, and especially the ACL, are among the most commonly injured structures of the knee. Given the increasing injury prevalence, there is undoubtedly a growing need for clinical decision making of health care providers. We reviewed the literature to analyze the diagnostic accuracy of the clinical examination for assessing ACL ruptures. METHODS AND MEASURES: MEDLINE (1966 to April 2005), EMBASE (1989 to April 2005), and CINAHL (1982 to April 2005) searches were performed. Also reference lists of the included studies were reviewed. Studies selected for data extraction were those that addressed the accuracy of at least 1 physical diagnostic test for ACL rupture and compared the performance of the clinical examination of the knee with a reference standard, such as arthroscopy, arthrotomy, or MRI. Searching was limited to English, German, and Dutch languages. RESULTS: Twenty-eight studies that assessed the accuracy of clinical tests for diagnosing ACL ruptures met the inclusion criteria. Study results were, however, heterogeneous. The Lachman test is the most valid test to determine ACL tears, showing a pooled sensitivity of 85% (95% confidence interval [CI], 83-87) and a pooled specificity of 94% (95% CI, 92-95). The pivot shift test is very specific, namely 98% (95% CI, 96-99), but has a poor sensitivity of 24% (95% CI, 21-27). The anterior drawer test shows good sensitivity and specificity in chronic conditions, respectively 92% (95% CI, 88-95) and 91% (95% CI, 87-94), but not in acute conditions. CONCLUSION: In case of suspected ACL injury it is recommended to perform the Lachman test. Because the pivot shift test is very specific both in acute as well as in chronic conditions, it is recommended to perform the pivot shift test as well. PMID- 16715829 TI - Relationships among lateral abdominal muscles, gender, body mass index, and hand dominance. AB - STUDY DESIGN: Exploratory. OBJECTIVES: To explore whether hand dominance, gender, and body mass index (BMI) influence the thickness of the lateral abdominal muscles as measured by ultrasound imaging. To document the extent of improvement in response stability when an average of multiple measures was utilized. BACKGROUND: Ultrasound imaging is a relatively new tool used to assess the lateral abdominal muscles. A better understanding of how these muscles contract in a healthy population can provide a reference for comparison to patients with low back pain (LBP). METHODS AND MEASURES: Thirty-two healthy participants (17 males, 15 females) aged 18 to 45 years (mean +/- SD, 31.9 +/- 7.8 years) were studied. Measurements of muscular thickness of the lateral abdominal muscles were obtained bilaterally while the subjects were at rest, and while they performed the abdominal drawing-in maneuver. To determine the possible influence of hand dominance and gender on muscle thickness, t tests were used. Correlation coefficients were used to assess the relationship between BMI and muscle thickness. Standard error of the measurement was used to assess response stability of the ultrasound imaging technique. RESULTS: No differences in the thicknesses of the transversus abdominis (TrA) muscle were measured during rest or while contracted, based on hand dominance (P > or = .73). Men had greater muscular thickness (P<.01), while the TrA in women represented a greater proportion of the total lateral abdominal muscles (P<.01). BMI was positively associated with muscle thickness (r> or =.66). Compared to a singular measurement, response stability improved by greater than 50% when an average of 3 measurements was used. CONCLUSIONS: Future researchers should assess the need to control for gender and BMI as potential covariates in ultrasound imaging studies of the lateral abdominal muscles. Asymmetry in the lateral abdominal muscles in those with LBP would be in direct contrast to the bilateral symmetry measured in those without LBP. PMID- 16715830 TI - Early application of negative work via eccentric ergometry following anterior cruciate ligament reconstruction: a case report. AB - STUDY DESIGN: Case report. OBJECTIVES: To present a progressively increasing negative-work exercise program via eccentric ergometry early after anterior cruciate ligament reconstruction (ACL-R) and to suggest the potential of negative work to amplify the return of quadriceps size and strength. CASE DESCRIPTION: The patient was a 26-year-old highly active recreational athlete who sustained an ACL tear while skiing in January 2004 and then again while skiing in February 2005. This individual underwent an arthroscopically assisted ACL-R with a double-loop semitendinosusgracilis autograft initially, then a patellar tendon autograft following his ACL graft rupture. Beginning within 3 weeks after surgery, a progressive negative-work exercise program was initiated using an eccentric ergometer. The patient completed 31 training sessions of 5 to 30 minutes in duration over a 12-week period following the ACL-R and 33 training sessions of the same frequency and duration following the ACL revision. OUTCOMES: Following ACL-R, quadriceps volume increased 28% (involved lower extremity) and 14% (uninvolved lower extremity) during the 12-week training program. Following revision, quadriceps volume returned to similar levels at the same postoperative period as those achieved after the initial surgery (2% less on the involved side and 2% greater on the uninvolved side). Quadriceps strength, 15 weeks after ACL R, exceeded preoperative measures by an average of 20% (involved) and 14% (uninvolved). Quadriceps strength after ACL revision exceeded all previous measures. DISCUSSION: This case report suggests that if gradually and progressively applied, negative work via eccentric ergometry can be both safe and efficacious early after ACL-R. Eccentric exercise may mitigate the prevalent muscle size and strength deficits commonly observed after ACL-R. The results of this case suggest a need for continued research with early negative work interventions following ACL-R. PMID- 16715831 TI - Plyometric exercise in the rehabilitation of athletes: physiological responses and clinical application. AB - Plyometric exercise was initially utilized to enhance sport performance and is more recently being used in the rehabilitation of injured athletes to help in the preparation for a return to sport participation. The identifying feature of plyometric exercise is a lengthening of the muscle-tendon unit followed directly by shortening (stretch-shortening cycle). Numerous plyometric exercises with varied difficulty and demand on the musculoskeletal system can be implemented in rehabilitation. Plyometric exercises are initiated at a lower intensity and progressed to more difficult, higher intensity levels. The progression to higher intensity plyometric exercise is thought to resolve postinjury neuromuscular impairments and to prepare the musculoskeletal system for rapid movements and high forces that may be similar to the demands imposed during sport participation, thus assisting the athlete with a return to full function. While there is a large body of scientific literature that supports the use of plyometric exercise to enhance athletic performance, evidence is sparse regarding the effectiveness of plyometric exercise in promoting a quick and safe return to sport after injury. This review will describe the mechanisms involved in plyometric exercise, discuss the considerations for implementing plyometric exercise into rehabilitation protocols, examine the evidence supporting the use of plyometric exercises, and make recommendations for future research. PMID- 16715832 TI - Effect of transducer velocity on intramuscular temperature during a 1-MHz ultrasound treatment. AB - STUDY DESIGN: A 3 x 2 repeated-measures design was used. The independent variables were transducer velocity (2-3 cm/s, 4-5 cm/s, and 7-8 cm/s) and time (pretreatment and posttreatment). OBJECTIVE: To determine if transducer velocity of a 1-MHz ultrasound treatment affects intramuscular tissue temperature. BACKGROUND: Most authors advocate ultrasound transducer velocities of 2 to 4 cm/s within an area of 2 to 3 times the effective radiating area or 2 times the size of the transducer head. However, a much faster rate of application (approximately 7-8 cm/s) is often observed in clinical settings. METHODS AND MEASURES: Eleven healthy screened volunteers (9 males, 2 females; mean +/- SD age, 22.6 +/- 1.7 years; mean +/- SD height, 175.7 +/- 13.7 cm; mean +/- SD body mass, 82.5 +/- 19.5 kg) were randomly assigned to a treatment order with all conditions administered during a single testing session. Each transducer velocity condition was administered for 10 minutes, using 1-MHz ultrasound with a 100% continuous duty cycle at an intensity of 1.5 W/cm2 over an area twice the size of the transducer head. After the first treatment, the 2 remaining subsequent velocity conditions were administered after the intramuscular temperature returned to within +/- 0.3 degrees C of the initial pretreatment temperature for 5 minutes. The dependent variable was left triceps surae muscle temperature measured at 3 cm below one half the measured skinfold thickness. RESULTS: Temperature increase across the 3 velocities was within 0.4 degrees C (F2.20 = 0.07, P = .93). Posttreatment values (mean +/- SD) ranged from 42.7 degrees C +/- 2.3 degrees C for the slowest velocity to 43.1 degrees C +/- 1.4 degrees C for the fastest velocity. Temperature increase was significant for time (F1.01 = 155.68, P<.00001), increasing from 37.8 degrees C +/- 0.8 degrees C pretreatment to 42.9 degrees C +/- 1.9 degrees C after treatment. CONCLUSION: Very similar intramuscular temperature increases can be observed among ultrasound treatments (10-minute duration, 1-MHz frequency, 100% continuous duty cycle, 1.5 W/cm2 intensity, within an area twice the size of the transducer head), with transducer velocities of 2 to 3, 4 to 5, and 7 to 8 cm/s. PMID- 16715833 TI - Descriptive report of shoulder range of motion and rotational strength 6 and 12 weeks following rotator cuff repair using a mini-open deltoid splitting technique. AB - STUDY DESIGN: Retrospective chart review. OBJECTIVES: To measure short-term postsurgery glenohumeral internal rotation and external rotation strength, shoulder range of motion (ROM), and subjective self-report ratings following mini open rotator cuff repair of full-thickness rotator cuff tears. BACKGROUND: Physical therapists provide rehabilitation for patients following mini-open rotator cuff repair. Long-term outcome studies have reported a high percentage of good and excellent results following surgery; however, little has been published regarding the immediate short-term results of this procedure, during which the patient is under the direct care of the physical therapist. MATERIALS AND METHODS: Charts from 11 female and 26 male patients, with a mean +/- SD age of 57.3 +/- 9.9 years, were reviewed following rotator cuff repair, using an arthroscopically assisted mini-open deltoid-splitting approach. All patients underwent postsurgery rehabilitation by the same therapist using a standard protocol. Retrospective chart review was used to obtain descriptive profiles of shoulder joint ROM at 6 and 12 weeks postsurgery and isokinetically assessed shoulder strength at 12 weeks postsurgery. RESULTS: For the postsurgical shoulder, ROM deficits ranging between 5 degrees to 7 degrees were measured for shoulder abduction and external rotation and internal rotation at 90 degrees of abduction. The postsurgical extremity had greater flexion ROM (9 degrees ) compared to the contralateral side. Isokinetic external rotation strength deficits of 5% to 7% were present at 12 weeks postsurgery, with 2% to 11% greater internal rotation shoulder strength on the operative extremity, when compared to the other side. Patients completed the self-report section of the modified American Shoulder Elbow Surgeons (ASES) Rating Scale at 12 weeks postsurgery and scored a mean of 38.7/45.0 points. CONCLUSION: The application of early ROM and progressive strengthening following mini-open rotator cuff repair allows for the successful return of ROM and strength 12 weeks postsurgery. The results of this study provide objective data for both shoulder ROM and strength at time points during which patients are traditionally receiving physical therapy following surgery. PMID- 16715834 TI - Applications of pooled DNA samples to the assessment of population affinities: short tandem repeats. AB - Pooled DNA samples have been used in association studies of Mendelian disease genes. This method involves combining equal quantities of DNA from patients and control subjects into separate pools and comparing the pools for distributions of genetic markers. In this study identical quantities of DNA from 300 individuals representing 6 populations were pooled and amplified for 296 loci using the touchdown polymerase chain reaction (PCR) method. The purpose of this study is to test the efficacy of pooled DNA markers in the reconstruction of the genetic structure of human populations. The populations sampled included Chuvash, Buryats, Kizhi, Native Americans, South Africans, and New York City whites. To test the accuracy of the allele-frequency distributions, we genotyped the Buryats and New York samples individually for six microsatellite markers and compared their frequencies to the allele frequencies derived from the electropherogram peak heights for the pooled DNA, producing a correlation of 0.9811 with a variance of less than 0.04. Two-dimensional scaling of genetic distances among the six populations produced clusters that reflected known historical relationships. A distance matrix was created using all 296 loci, and matrices based on individual chromosomes were correlated against the total matrix. As expected, the largest chromosomes had the highest correlations with the total matrix, whereas one of the smallest chromosomes, chromosome 22, had the lowest correlation and differed most from the combined STR distance matrix. PMID- 16715835 TI - Mandibular symphysis of large-bodied hominoids. AB - The hominoid mandibular symphysis has received a great deal of attention from anatomists, human biologists, and paleontologists. Much of this research has focused on functional interpretations of symphyseal shape variation. Here, we examine the two-dimensional cross-sectional shape of the adult mandibular symphysis for 45 humans, 42 chimpanzees, 37 gorillas, and 51 orangutans using eigenshape analysis, an outline-based morphometric approach. Our results demonstrate that a large proportion of the variation described by the first eigenshape correlates with proposed functional adaptations to counteract stresses at the mandibular midline during mastication. Subsequent eigenshapes describe subtle aspects of shape variation in the mandibular symphysis. The morphology associated with these eigenshapes does not conform with functional predictions, nor does it show a relationship with sexual dimorphism. However, eigenshapes provide for considerable taxonomic discrimination between the four taxa studied and may consequently prove useful in the analysis of fossil material. Comparison with elliptical Fourier analysis of the mandibular symphysis identifies eigenshape analysis as providing superior taxonomic discrimination. The results presented here demonstrate that the cross-sectional shape of the mandibular symphysis results from a complex interplay of functional and nonfunctional influences and for the first time identifies and quantifies the specific aspects of variation attributable to these factors. PMID- 16715836 TI - Growth of Jersey schoolchildren during the 1940-1945 German occupation: comparison with schoolchildren on mainland Britain. AB - The 1940-45 German occupation of the Channel Islands was accompanied by a gradual deterioration in food availability and culminated in a nine-month siege with severe food shortages after supplies from France were cut off following the liberation of Normandy in 1944. The aim of this study was to compare the growth rates of schoolchildren resident on one of the Channel Islands (Jersey) with children growing up on mainland Britain. Secondary analyses were conducted on published data describing the growth of approximately 3500 schoolchildren resident on Jersey throughout the occupation, 10,000-15,000 London schoolchildren, and about 5000 children evacuated from mainland cities to "residential camp schools" in rural areas. A comparison of age- and sex-adjusted average annual growth rates between 1940 and 1943 confirmed that 8 1/2 - 12 1/2 year-old children on Jersey gained 28.2-33.1% less weight and 23.6-26.5% less height per year than children on the mainland. Because food shortages on Jersey intensified after 1943 (when the mid-occupation growth measurements had been recorded), growth rates toward the end of the occupation (in 1944 and early 1945) are likely to have been even worse than those observed between 1940 and 1943. As such, these analyses provide a somewhat conservative estimate of the decline in childhood growth rates that occurred on Jersey during the 1940-45 German occupation. PMID- 16715837 TI - Genotype-by-sex interaction in the regulation of high-density lipoprotein: the Framingham Heart Study. AB - Low levels of high-density lipoprotein (HDL) are widely documented as a risk factor for cardiovascular disease (CVD). Furthermore, there is marked sexual dimorphism in both HDL levels and the prevalence of CVD. However, the extent to which genetic factors contribute to such dimorphism has been largely unexplored. We examined the evidence for genotype-by-sex effects on HDL in a longitudinal sample of 1562 participants from 330 families in the Framingham Heart Study at three times points corresponding approximately to 1971-1974, 1980-1983, and 1988 1991. Using a variance component method, we conducted a genome scan of HDL at each time point in males and females, separately and combined, and tested for genotype-by-sex interaction at a quantitative trait locus (QTL) at each time point. Consistent findings were noted only for females on chromosome 2 near marker D2S1328, with adjusted LOD scores of 2.6, 2.2, and 2.1 across the three time points, respectively. In males suggestive linkage was detected on chromosome 16 near marker D16S3396 at the second time point and on chromosome 18 near marker D18S851 at the third time point (adjusted LOD = 2.2 and 2.4, respectively). Although the heritability of HDL is similar in males and females, sex appears to exert a substantial effect on the QTL-specific variance of HDL. When genotype-by sex interactions exist and are not modeled, the power to detect linkage is reduced; thus our results may explain in part the paucity of significant linkage findings for HDL. PMID- 16715838 TI - Beta 3 adrenergic receptor Trp64Arg polymorphism and manifestation of coronary artery disease in Arabs. AB - The substitution of tryptophan (Trp) by arginine (Arg) at position 64 in the beta3-adrenoceptor (beta3-AR) gene has been associated with obesity, diabetes mellitus, and coronary artery disease (CAD). We have investigated whether the Trp64Arg polymorphism is associated with the manifestation of CAD or one of its important risk factors, such as obesity, diabetes mellitus, elevated cholesterol and triglyceride levels, or hypertension in the Arab population. All participating subjects were genotyped for this polymorphism using the polymerase chain reaction followed by enzymatic digestion and sequencing. In the angiographed normal control subjects (n=495), 90.3% were homozygous Trp/Trp, 9.5% were heterozygous Trp/Arg, and 0.2% were homozygous for the Arg/Arg genotype, compared to 87%, 12.3%, and 0.7%, respectively, among angiographically confirmed CAD patients (n=981). There was no statistical difference in the distribution of genotypes or allele frequencies between the CAD and control groups. We carried out a stepwise logistic regression analysis to study the possible combined effect of the genotypes and other risk factors on CAD. All variables were retained in the model, with p values of 0.014, 0.006, 0.005, < 0.001, 0.045, 0.002, < 0.001, and 0.016 for genotype, diabetes mellitus, sex, family history of CAD, obesity, myocardial infarction, smoking, and age, respectively. In conclusion, the Trp64Arg polymorphism of the beta3-AR gene does not represent an independent risk factor for CAD in Arabs. However, in the presence of other CAD risk factors, this polymorphism may be used as a predictor of CAD. PMID- 16715839 TI - Microsatellite diversity in Andhra Pradesh, India: genetic stratification versus social stratification. AB - DNA samples of 948 individuals belonging to 27 populations from southern Andhra Pradesh were analyzed for nine AmpFlSTR Profiler Plus loci. The nature and extent of genomic diversity within and between these populations have been examined with reference to socioeconomic and geographic affiliations. The results suggest that the average heterozygosity is uniformly high in these populations (> 0.80) and that the patterns of allele distributions are similar across the populations. The value of the coefficient of gene differentiation and the AMOVA and structure analysis results suggest that these populations are highly homogeneous. The neighbor-joining tree constructed using either D(A) or F(ST) distances suggests no intelligible pattern of population clusters based on ethnohistoric or geographic affiliations. All these observations suggest either a common recent origin of these populations or extensive gene flow across the populations that erased the original genetic differences. Given strict endogamy, the latter explanation can hold only if there has been unauthorized or unrecognized gene flow transecting the social boundaries. Nevertheless, the regression plot of average heterozygosity versus distance from the centroid (Rii), based on Harpending and Ward's (1982) model, and the genetic distances computed between different hierarchical groups within Andhra Pradesh tend to support this conjecture. Overall, the results suggest lack of a significant degree of genetic stratification that is consistent with social stratification in Andhra Pradesh. Furthermore, the neighbor-joining tree based on comparative data from other Indian and continental populations brings out a single and compact cluster of all the Andhra populations that is clearly separated from the rest. PMID- 16715840 TI - Autosomal STR variation in five Austronesian populations. AB - Human population characteristics at the genetic level are integral to both forensic biology and population genetics. This study evaluates biparental microsatellite markers in five Austronesian-speaking groups to characterize their intra- and interpopulation differences. Genetic diversity was analyzed using 15 short tandem repeat (STR) loci from 338 unrelated individuals from 5 Pacific islands populations, including the aboriginal Ami and Atayal groups from Taiwan, Bali and Java in Indonesia, and the Polynesian islands of Samoa. Allele frequencies from the STR profiles were determined and compared to other geographically targeted worldwide populations procured from recent literature. Hierarchical AMOVA analysis revealed a large number of loci that exhibit significant correspondence to linguistic partitioning among groups of populations. A pronounced divide exists between Samoa and the East (Formosa) and Southeast Asian (Bali and Java) islands. This is clearly illustrated in the topology of the neighbor-joining tree. Phylogenetic analyses also indicate clear distinctions between the Ami and Atayal and between Java and Bali, which belie the respective geographic proximities of the populations in each set. This differentiation is supported by the higher interpopulation variance components of the Austronesian populations compared to other Asian non-Austronesian groups. Our phylogenetic data indicate that, despite their linguistic commonalities, these five groups are genetically distinct. This degree of genetic differentiation justifies the creation of population-specific databases for human identification. PMID- 16715841 TI - Haplotype distribution of and linkage disequilibrium between four polymorphic markers near the CFTR locus in Brazilian cystic fibrosis patients. AB - To contribute to a better understanding of the origin and distribution of CFTR mutations in the Brazilian population, we have investigated the linkage between four polymorphic markers (XV2c, KM19, GATT, and TUB9) within or near the CFTR locus. The distribution of alleles for each polymorphism for both parental and cystic fibrosis (CF) chromosomes from Rio de Janeiro CF families were ascertained using a maximum-likelihood method. This same method was applied to study the distribution of the haplotypes defined by these markers. There was no significant association between the XV2c and KM19 loci on the parental and CF chromosomes. On the other hand, a strong association between GATT and TUB9 loci was observed on both CF and parental chromosomes, and striking linkage disequilibrium between the GATT-TUB9 pair and deltaF508 was observed (chi2 = 26.48, p < 0.0001). Remarkable linkage disequilibrium between the GATT-TUB9 marker pair and non-deltaF508 was also found (chi2 = 17.05, p < 0.0001). Our finding of a linkage disequilibrium between GATT-TUB9 and the CFTR locus could suggest that gene flow between different ethnic groups, mainly sub-Saharan and Mediterranean populations, with Brazilian populations could have resulted in some CF mutations originating on chromosomes that carried the GATT-TUB9 marker haplotype 7-2 (OR = 1.34 < 2.83 < 6.00; p = 0.0066). PMID- 16715842 TI - Distribution of apolipoprotein E alleles in coras and huicholes from Nayarit and Nahuas and Mestizos from Veracruz, Mexico. AB - We report allele frequencies for the most common polymorphism of the APOE gene in Mexican individuals from two regions not previously described: Coras and Huicholes from Nayarit, and Nahuas and mestizos from Veracruz. We also report APOE allele frequencies for inhabitants of Mexico City. These descriptive data underscore the allelic heterogeneity for this particular locus in Mexico. PMID- 16715844 TI - [Save staffing saves lives!]. PMID- 16715843 TI - Frequencies of the four major Amerindian mtDNA haplogroups in the population of Montevideo, Uruguay. AB - mtDNA Amerindian polymorphisms were studied in 108 inhabitants of Montevideo, Uruguay, using PCR RFLP analysis. Amerindian haplogroups were found in 20.4% of the sample. The frequency of Amerindian polymorphisms in Montevideo differed significantly from that observed in Tacuarembo, a city about 400 km away, indicating the high level of variation within Uruguay. Results for mitochondrial markers indicate that admixture occurred primarily as a result of Amerindian females mating with European males. PMID- 16715845 TI - [New studies give the first results on geriatric nursing continuing education in Germany]. PMID- 16715846 TI - [Personal relations is our capital in the future]. PMID- 16715848 TI - [At the beginning, matters are comfortable]. PMID- 16715847 TI - [Saving on staff personnel--shortage of nurses in geriatric nursing]. PMID- 16715849 TI - [Talking instead of writing]. PMID- 16715850 TI - [After care and preparedness. . Ambulance nursing service]. PMID- 16715851 TI - [Nurse practitioner or specialist?]. PMID- 16715852 TI - [World Health Day. People promote health--the health professions]. PMID- 16715853 TI - [A problem of an unsteady financial economy?]. PMID- 16715854 TI - [A good memory is the key to the future]. PMID- 16715855 TI - [Work-related colleagues]. PMID- 16715856 TI - [Who connects with ambulance nurses?]. PMID- 16715857 TI - [Working until the age of 67]]. PMID- 16715858 TI - [No hurdles in geriatric nursing]. PMID- 16715859 TI - [Dyspnoea caused by upper-airway obstruction: simple diagnosis by establishing a flow-volume loop]. AB - Three patients, men aged 72, 78 and 19 years, experienced shortness of breath and laboured breathing. All three had an upper-airway obstruction detected by a flow volume loop and confirmed by bronchoscopy. The first patient had oesophageal carcinoma with vocal-cord paralysis and soon died. The second patient had a large struma; flow-volume loop improved after strumectomy. The third patient was diagnosed with extragonadal testicular carcinoma. The flow-volume loop improved after the first chemotherapy session. Flow-volume loop is an easy, non-invasive diagnostic tool that can be used even in severely-ill patients. It can provide information about the location of the obstruction and can differentiate between obstructive pulmonary disease and upper-airway obstruction. Therefore, it is recommended to obtain a flow-volume loop during the assessment of patients with upper airway obstruction. PMID- 16715860 TI - [The recommended reference values for anaemia in the elderly are of limited usefulness]. AB - Recent epidemiological findings on the risk factors for mortality and functional impairment do not support lower cut-off values than the official WHO reference values for the definition of anaemia in the elderly, and even suggest higher limits. However, wide-scale application of such high Hb values for a full investigation of the causes of anaemia in clinical practice requires data on the effectiveness in terms of a net increase in quality of life at acceptable costs. At present, there are no evidence-based Hb limits for the elderly because such data are lacking. As a pragmatic way out, the WHO limits can be taken for a simple and non-invasive biochemical analysis of anaemia, but individual considerations, taking into account co-morbidity, life expectation and patients' preferences, are necessary for further diagnostic investigations such as colonoscopy or sternum puncture. PMID- 16715861 TI - [Reference values for anaemia in the elderly]. AB - According to the reference values of the World Health Organization (WHO), anaemia is defined as a haemoglobin (Hb) level < 8.1 mmol/l in men and < 7.5 mmol/l in women. Lower reference values are often used in older patients because many doctors consider a decreased Hb level to be physiological in advanced age. However, Hb values under the WHO reference values are associated with increased mortality in older persons. Moreover, a cause of the anaemia is found in > 80% of older persons with anaemia if further diagnostic tests are done. Decreased Hb levels are also associated with functional impairments such as mobility disorders. These findings apply not only to severely decreased Hb levels but also to Hb levels that are only slightly below the WHO reference values. Correspondingly, increased erythropoietin levels are seen in older persons with severely decreased Hb levels as well as in older persons with slightly decreased Hb levels. Thus, a decrease in the Hb level in advanced age is not physiological and the WHO reference values for anaemia are also valid for older persons. PMID- 16715862 TI - [Diabetes mellitus: simple classification and corresponding treatment]. AB - The classification of diabetes mellitus is not always straightforward but is of clinical importance in view of the differences in treatment, prognosis and inheritance. In young patients with acute signs ofhyperglycaemia and a low bodymass index (BMI), diabetes type 1 is the evident diagnosis, and insulin therapy should be initiated as quickly as possible. Deafness in patients with mild symptoms and a highly positive family history suggests the presence of a mitochondrial or maternally inherited type of diabetes: maternally inherited diabetes and deafness (MIDD). In the absence of deafness, maturity onset diabetes of the young (MODY) should be suspected in the same group of patients. In case of a non-acute presentation of hyperglycaemia without an evident family history of diabetes, the BMI can be helpful in the next steps of the diagnostic process. In practice, the proposed flow chart can readily be applied for the classification and treatment of patients with hyperglycaemia. PMID- 16715863 TI - [Diagnostic image (273). A boy unable to walk]. AB - A 16-year-old boy suffered from progressive paralysis and retching when attempting to drink, due to rabies. PMID- 16715864 TI - [Market forces inevitable in health care]. AB - Market forces inevitable in health care. - Free markets exist everywhere and have provided us, as a society, with many benefits. A truly free market does not exist: all market sectors are bound within legal frameworks. For health care, the question is not whether a free market is feasible, but within which frameworks does it work exclusively. Establishing the framework is a particular political responsibility. Without the framework, health care will fall prey to political opportunists and lobbyists for pressure groups. PMID- 16715865 TI - [A free market is unfavourable for health care]. AB - The new health care system in the Netherlands introduces a free market into the health care sector. Because health care lacks some essential conditions, a free market will not provide effective resource allocation in this sector. In addition, a free market leads to more bureaucracy and less solidarity, cooperation and freedom of choice. It also impairs the autonomy of the medical professional. PMID- 16715866 TI - [Use of rofecoxib in family practice in the north of Netherlands, 2000-2004: background and consequences]. AB - OBJECTIVE: To gain insight into the trends in the prescription ofrofecoxib, the medication and disease histories of rofecoxib users, and the occurrence of cardiac complications during the follow-up. DESIGN: Longitudinal comparative study in family practice. METHOD: Based on the data from family practices (17 family physicians) affiliated with the Registration Network Groningen, a network with about 30,000 patients in the northern part of the Netherlands, the incidence and prevalence ofrofecoxib use were determined per 1000 patients at risk for every quarter during the period 2000-2004. Data on duration, dosage and indications were taken from the rofecoxib prescriptions. Differences in morbidity between rofecoxib users and the users of traditional NSAIDs were compared using logistic regression, and differences between the two groups in the occurrence of cardiac complications were assessed by means of Cox regression analysis. RESULTS: During the period investigated, rofecoxib was prescribed with increasing frequency in family practice; there was a total of 1784 prescriptions for 509 patients. The drug was used increasingly for short durations and for a growing number of indications. Rofecoxib was selectively prescribed to former users of traditional NSAIDs. The occurrence of acute myocardial infarction, stroke or 'transient ischaemic attack' in the follow-up period was associated especially with pre-existing cardiovascular disease, but these complications were also seen more often among patients without pre-existing cardiovascular disease who used rofecoxib than among those who used the traditional NSAIDs (the difference was not significant). CONCLUSION: The observations indicate that, via a process of channelling, rofecoxib was prescribed to a highly-specific population of patients who, paradoxically, were at a higher risk of developing the same cardiac complications with which rofecoxib had been shown to be associated in randomised clinical trials. PMID- 16715867 TI - [Colistin as last resort in 2 patients with pulmonary infection with multiresistant nosocomial bacteria]. AB - A 57-year-old man requiring ventilation after gastric-tube reconstruction (due to oesophageal carcinoma) and a 16-year-old boy with thoracic trauma developed pneumonia caused by Pseudomonas aeruginosa and other pathogens. Their infections persisted during treatment with antibiotics. The antibiotic colistin was added and the clinical condition improved in both patients after 3-4 weeks. Eventually, they both were transferred to a rehabilitation clinic. The use of colistin was abandoned in the 1970s due to alleged nephro- and neurotoxicity. In recent observational studies, the drug appeared to cause fewer toxic side-effects than previously thought. Colistin is only registered for inhaled use for lung infections and for resistant pulmonary infections in patients with cystic fibrosis, but it may also be effective in the treatment of other patients with multidrug-resistant, Gram-negative bacteria. PMID- 16715868 TI - [Life-threatening ventricular tachycardia during flecainide treatment for symptomatic atrial fibrillation in a patient with a structural cardiac disorder]. AB - A 37-year-old man with symptomatic acute atrial fibrillation and a low-voltage electrocardiogram was treated with flecainide intravenously. Instead of conversion to sinus rhythm, he developed a wide-complex tachycardia suggestive of ventricular tachycardia. The patient recovered following electric cardioversion. First-choice therapy for symptomatic atrial fibrillation of recent onset (duration < 48 hours) is chemical conversion with a class IC antiarrhythmic drug (e.g. flecainide, propafenone). However, in patients with structural heart disorders, these drugs may induce ventricular tachycardia. A low-voltage electrocardiogram is suggestive of left ventricular damage. For these patients, electric cardioversion is a better alternative. PMID- 16715870 TI - [Fatal asthma in childhood preventable by recognizing risk factors and presenting features]. PMID- 16715871 TI - [Fatal asthma in childhood preventable by recognizing risk factors and presenting features]. PMID- 16715872 TI - [Fatal asthma in childhood preventable by recognizing risk factors and presenting features]. PMID- 16715873 TI - [Perinatal mortality in the Netherlands: everyone's problem and yet no one's problem]. PMID- 16715874 TI - The genus Hyalomma Koch, 1844. i. reinstatement of Hyalomma (euhyalomma) glabrum Delpy, 1949 (Acari, Ixodidae) as a valid species with a redescription of the adults, the first description of its immature stages and notes on its biology. AB - For nearly 50 years the ixodid tick Hyalomma marginatum turanicum, reputedly introduced into South Africa on imported Persian sheep, has been considered identical to the Asian Hyalomma (Euhyalomma) marginatum turanicum Pomerantzev, 1946. Comparisons of this tick with the Asian H. (E.) m. turanicum and other subspecies of Hyalomma (Euhyalomma) marginatum, however, reveal that it is an old taxon, namely Hyalomma rufipes glabrum Delpy, 1949. It is hereby reinstated as Hyalomma (Euhyalomma) glabrum, and its adults are redescribed and its immature stages described for the first time. The preferred hosts of its adults are large herbivores such as zebras, gemsbok and eland, on which it occurs during summer. The preferred hosts of its immature stages are scrub hares and ground-frequenting birds, on which it is present during autumn and winter. Data on its distribution and possible disease relationships are also provided. PMID- 16715875 TI - Hosts, seasonality and geographic distribution of the South African tortoise tick, Amblyomma marmoreum. AB - The tortoise tick Amblyomma marmoreum was collected from large numbers of reptiles and other animals during the course of numerous surveys conducted in South Africa. A total of 1229 ticks, of which 550 were adults, were recovered from 309 reptiles belonging to 13 species, with leopard tortoises, Geochelone pardalis being the most heavily infested. The 269 birds sampled harboured 4901 larvae, 217 nymphs and no adult ticks, and the prevalence of infestation was greatest on helmeted guineafowls, Numida meleagris. Only two larvae were recovered from 610 rodents, including 31 spring hares, Pedetes capensis, whereas 1144 other small mammals yielded 1835 immature ticks, of which 1655 were collected from 623 scrub hares, Lepus saxatilis. The 213 carnivores examined harboured 2459 ticks of which none were adult. A single adult tick and 6684 larvae and 62 nymphs were recovered from 656 large herbivores, and a total of 4081 immature ticks and three adults were collected from 1543 domestic animals and 194 humans. Adult male and female A. marmoreum were most numerous on reptiles during January and February, and larvae during March. The largest numbers of larvae were present on domestic cattle and helmeted guineafowls in the Eastern Cape Province during March or April respectively, whereas larvae were most numerous on helmeted guineafowls, scrub hares and the vegetation in north-eastern Mpumalanga Province during May. In both provinces nymphs were most numerous between October and December. Amblyomma marmoreum appears to be most prevalent in the western regions of the Western and Eastern Cape and Free State provinces, and the north-eastern regions of the Northern Cape, KwaZulu-Natal, Mpumulanga and Limpopo provinces. PMID- 16715876 TI - Check-list of the pentastomid parasites crocodilians and freshwater chelonians. AB - Based on published records and own data a summary is given of the geographical distribution of the currently known species of pentastomid parasites infecting crocodiles and alligators, as well as freshwater chelonians. A brief generic diagnosis is provided for each genus. Fourteen out of the currently 23 living crocodilian species have been recorded as being host to one or more pentastomes. Out of the 32 pentastome species six are considered species inquirendae. Presently, six genera of crocodilian pentastomes, Agema, Alofia, Leiperia, Sebekia, Selfia and Subtriquetra are recognized. African crocodiles harbour eight pentastome species, six of which have been recorded from the Nile crocodile, Crocodylus niloticus. Three species belong to the genus Sebekia, Alofia being represented by two and Leiperia by only one species. Two species, Alofia parva and Agema silvae-palustris, occur in the dwarf crocodile, Osteolaemus tetraspis, and the slender-snouted crocodile, Crocodylus cataphractus, exclusively, but a single Sebekia species is shared with the Nile crocodile. The genus Agema is endemic to the African region. Infective stages of the pentastome Subtriquetra rileyi, thought to utilize Nile crocodiles as final hosts, have been recovered only from fishes. The largest number of pentastome species is found in the Australasian region. Of these, the Indo-Pacific crocodile, Crocodylus porosus, harbours seven, representing the genera Alofia, Sebekia, Leiperia and Selfia. Selfia is exclusive to the latter host. The genus Subtriquetra has been reported from "Indian crocodiles", a term possibly referring to either Crocodylus palustris, Crocodylus porosus or Gavialis gangeticus. Ten species of pentastomes parasitizing the crocodilian genera Alligator, Caiman, Crocodylus and Melanosuchus have been recorded from the Neotropical region including the southern states of the North American continent. The two most wide-spread pentastome genera, Alofia and Sebekia, have been recorded together with representatives of the genus Subtriquetra and immature and larval forms of Leiperia. To date the two monospecific genera, Pelonia, from two terrapin species, Pelusios sinuatus and Pelomedusa subrufa, in South Africa, and Diesingia from Hydraspis geoffroyana and Hydromedusa tectifera in South America, are the only chelonian pentastomes recovered world-wide. A possible exception is the crocodilian pentastome Sebekia mississippiensis which can reach maturity in experimentally infected terrapins. PMID- 16715877 TI - Epidemiological studies of Fasciola gigantica infections in cattle in the highveld and lowveld communal grazing areas of Zimbabwe. AB - During the period between January 1999 and December 2000, the distribution and seasonal patterns of Fasciola gigantica infections in cattle in the highveld and lowveld communal grazing areas of Zimbabwe were determined through monthly coprological examination. Cattle faecal samples were collected from 12 and nine dipping sites in the highveld and lowveld communal grazing areas respectively. Patterns of distribution and seasonal fluctuations of the intermediate host-snail populations and the climatic factors influencing the distribution were also determined by sampling at monthly intervals for a period of 24 months (November 1998 to October 2000) in six dams and six streams in the highveld and in nine dams in the lowveld communal grazing areas. Each site was sampled for relative snail density and the vegetation cover and type, physical and chemical properties of water, and mean monthly rainfall and temperature were recorded. Aquatic vegetation and grass samples 0-1 m from the edges of the snail habitats were collected monthly to determine the presence or absence of F. gigantica metacercariae. Snails collected at the same time were individually checked for the emergence of larval stages of F. gigantica. A total of 16264 (calves 5418; weaners 5461 and adults 5385) faecal samples were collected during the entire period of the study and 2500 (15.4%) of the samples were positive for F. gigantica eggs. Significantly higher prevalences were found in the highveld compared to the lowveld (P < 0.001), for adult cattle than calves (P < 0.01) and in the wet season over the dry season (P < 0.01). Faecal egg output peaked from August/September to March/April for both years of the study. Lymnaea natalensis, the snail intermediate host of F. gigantica was recorded from the study sites with the highveld having a significantly higher abundance of the snail species than the lowveld (P < 0.01). The snail population was low between December and March and started to increase in April reaching a peak in September/October. The number of juvenile snails peaked between April and August. The mean number of snails collected was negatively correlated with rainfall and positively correlated with temperature. Mean number of snails collected was also positively correlated with Potamogeton plant species and negatively correlated with Cyperus plant species. However, none of the L. natalensis collected from the habitats were found shedding Fasciola cercariae. Metacercariae were found on herbage from the fringes of the snail habitats between February and August for both years, with most of the metacercariae concentrated on herbage 0-1 m from the banks of the habitats. Based on the findings of this study, anthelmintic treatment should be administered in December/January to control chronic and mature fasciolosis. A second treatment should be given in April/May to reduce pasture contamination and subsequently snail infection, as this is the time the snail population starts to build up. To control acute fasciolosis due to the immature liver flukes a third treatment should be given in August. The first application of molluscicides to control the snail intermediate hosts can be done in June the time when the snail is harbouring the parasite and a second application in September in order to kill new generations of infected snails PMID- 16715878 TI - Prevalence of common gastrointestinal nematode parasites in scavenging pigs of different ages and sexes in eastern centre province, Burkina Faso. AB - The range and infestation intensities of gastrointestinal parasitic nematode species depend on the type of swine production system. The present study focused mainly on nematodes of veterinary importance in scavenging pigs in Burkina Faso, and aimed at determining the prevalence of gastro-intestinal nematode parasites by means of faecal egg per gram (EPG) counts. Between November 2001 and October 2002, faecal samples from 383 pigs of different sexes and ages (< 5 months, 5-12 months and > 12 months) were collected from the rectum and examined for gastrointestinal nematodes parasites using the Mc Master method. Of the 383 pigs examined, 91% were infected by one or more parasites. Ascaris suum (40%; 100-1 400 EPG) was the most prevalent parasite followed by Strongyloides ransomi (21%; 100-4200 EPG), Oesophagostomum spp. (18%; 100-1000 EPG), Hyostrongylus rubidus (11%; 100-1 800 EPG), Globocephalus spp. (10%; 100-400 EPG) and Trichuris suis (1 %; 100-200 EPG). The prevalence was significantly higher in female pigs (n = 239) than in males. In addition, females excreted significantly (P < 0.05) more eggs in their faeces than males, except in the case of Globocephalus spp. The age of the animal had no effect on the prevalence of A. suum whereas there were significant differences in age categories concerning S. ransomi, H. rubidus, Oesophagostumum spp. and Globocephalus spp. Unexpectedly, the high prevalence of these common parasites was not accompanied by elevated EPG values, which suggests the existence of moderate infestations. The present work indicates that the common nematode infestations in pigs do not necessarily need a systematic herd anthelmintic treatment, as only a small number of worms is required to induce immunity. A further study is needed to formulate appropriate and cost-effective strategies for the control of gastro-intestinal nematode parasites in pigs in Burkina Faso. PMID- 16715879 TI - The effect of GnRH or oestradiol injected at pro-oestrus on luteal function and follicular dynamics of the subsequent oestrous cycle in non-lactating cycling Holstein cows. AB - Oestrous synchronization involves synchronization of ovarian follicular turnover, new wave emergence, and finally induction of ovulation. The final step can be synchronized by the parenteral administration of either GnRH or oestradiol benzoate. This study investigated corpus luteum and follicular emergence after ovulation had been induced by the administration of either GnRH or oestradiol benzoate. The injection of oestradiol benzoate may have delayed the emergence of the first follicular wave subsequent to the induced ovulation; administration of oestradiol benzoate or GnRH lowered the progesterone rise so that the maximum dioestrous concentration of progesterone on Day 9 was lower when cows were treated during pro-oestrus compared to the spontaneously ovulating controls. One implication of findings from the present study is that induction of ovulation with either oestradiol benzoate or GnRH, administered 24 or 36 h after withdrawal of the CIDR device, respectively, may lower fertility. Future studies must identify the timing of administration relative to the time of CIDR device withdrawal and the optimum concentration of oestradiol benzoate or GnRH that would not have untoward effects on the development of the corpus lutea, particularly within the first week of dioestrus. PMID- 16715880 TI - Faecal helminth egg and oocyst counts of a small population of African lions (Panthera leo) in the southwestern Kalahari, Namibia. AB - An endoparasite survey of a small pride of African lions (Panthera leo) was conducted at Intu Afrika Kalahari Game Reserve, southwestern Namibia, during winter and summer of 2003 and 2004, respectively. Overall, 23 fresh lion scats were collected opportunistically during fieldwork trials. A flotation technique was employed for the diagnosis of parasites. Three nematodes, Ancylostoma braziliense, Gnathostoma spinigerum and Uncinaria stenocephala and two coccidians, Toxoplasma gondii and Isospora felis were recorded. By using the McMaster method for quantification, a maximum number of 14866 oocysts per gram of faeces was obtained for I. felis during winter 2003. Endoparasite taxa carried by the different individuals in the pride were found to be related to their levels of association. Rates of infection were relatively low as a result of the habitat, semi-captive conditions and earlier sporadic deworming. PMID- 16715882 TI - [Clinical analysis of operative and hospital deaths in primary lung cancer]. AB - In our department, there were 313 thoracic surgeries for primary lung cancer from January 1994 to December 2003. We clinically reviewed for the operative and hospital death (n=18, 5.8%). The patients were 16 males and 2 females (70.6 +/- 5.6 years old). The surgical procedures were 4 pneumonectomies, 13 lobectomies (3 bronchoplasties) and 1 partial resection. The mean interval until postoperative death was 122.5 +/- 156.1 days. There were 5 direct operative deaths within 30 days (1.6%). There were 4 cancer deaths, 2 hemoptyses, 2 operative bleeding, 2 thromboses, 2 cerebral hemorrhages, 1 pyothorax, 1 pneumonia, 1 respiratory failure, 1 multiple organ failure after chemotherapy and 2 unexplained deaths. The patients with pneumonectomy or aged significantly had high mortality. For postoperative complications such as hemoptysis or bleeding, perioperative management that takes these issues into consideration is needed. Furthermore, we must carefully review the preoperative evaluation and combined treatment, because there were many cancer deaths among cases showing early recurrence and metastasis. PMID- 16715881 TI - An update of the bovine trypanosomosis situation at the edge of Hluhiuwe-Imfolozi Park, Kwazulu-Natal Province, South Africa. AB - To obtain updated data on and assess the contribution of trypanosomosis to the disease burden of cattle kept at the edge of the Hluhluwe-iMfolozi Park, a survey was conducted at Mvutshini Dip. Use was made of a purposeful sampling strategy by restricting sampling to animals that the livestock owner considered to be in poor condition. Of a total of 76 blood samples collected, 26 were parasitologically positive and 46 were positive on PCR/RFLP. Almost all infections were due to Trypanosoma congolense savannah subgroup. A total of 63 animals had a PCV < or = 24% and were considered to be anaemic. Results from the survey show that trypanosome infections contribute significantly to the overall burden of disease in the area. Further research is required to develop appropriate control methods. PMID- 16715883 TI - [Successful off-pump coronary artery bypass for a patient with aplastic anemia]. AB - We report a 61-year-old man with aplastic anemia who underwent successful off pump coronary artery bypass (OPCAB) after being admitted for angina pectoris. Coronary angiography showed severe stenosis of the left main coronary artery. Preoperative WBC was 2,200/microl, neutrophil 704/microl, Hb 8.1g/dl, and PLT 16,000/microl. We conducted OPCAB on double vessels using left internal thoracic and radial artery grafts. Thirty units of platelets were transfused intraoperatively with little perioperaive hemorrhage. Because of high grade fever, we injected 150 microg granulocyte colony-stimulating factor (G-CSF) every 3 days postoperatively to prevent major infection. The combination of appropriate perioperative management and OPCAB yielded an effective result for a patient with severe hematological disorders causing pancytopenia. PMID- 16715884 TI - [Pulmonary actinomycosis developing diffuse phregmone after pneumonectomy]. AB - The patient was a 75-year-old male who consulted the department of respiratory tract internal medicine in our hospital for left chest pain occurring from the beginning of December 2003. Chest X-ray indicated a tumorous shadow in the left lower lung field. A chest CT also revealed an irregularly shaped mass shadow in the left lower lobe. Since bronchoscopy failed to establish a definitive diagnosis, the patient was referred to our department for surgery to undertake thoracotomy. After left pneumonectomy being performed based on a suspicion of lung abscess, pathological examination of specimen from the resected left lung showed sulfur granules which led to the diagnosis of pulmonary actinomycosis. Because of the diffuse phregmone developing around the surgical wound, benzylpenicillin potassium administration was started, and was continued for a further 6 months on an outpatient basis. Pulmonary actinomycosis is a relatively rare chronic pulmonary infection. It is often difficult to distinguish pulmonary actinomycosis from other pulmonary disease such as lung cancer because of the similarity of their appearance on X-ray or CT, and almost all cases of pulmonary actinomycosis are diagnosed by thoracotomy. PMID- 16715885 TI - [Emergent coronary artery bypass grafting for a patient with cardiopulmonary arrest]. AB - A 59-year-old man was admitted to our hospital due to sudden onset of unconsciousness caused by myocardial infarction with ventricular fibrillation. Emergent coronary angiography under intraaortic balloon pumping revealed 90% stenosis of the left main trunk and left anterior descending artery (LAD), and complete obstruction of the left circumflex artery (Cx) and right coronary artery (RCA). Emergent coronary artery bypass grafting (CABG) to LAD, Cx, and RCA was performed. During the postoperative course, the patient developed ventricular tachycardia/fibrillation. After implantation of an implantable cardioverter defibrillator (ICD), he was discharged on the postoperative day 36. The patient has now resumed normal daily life. PMID- 16715886 TI - [Drug-eluting stents restenosis 2 months after the procedure and necessitating semi-emergency off-pump coronary artery bypass]. AB - Drug-eluting stents (sirolimus-eluting stent: Cypher stent) have showed a reducing the frequency of in-stent restenosis and a good safety profile. One case involving edge dissection with Cypher stent restenosis for 2 lesions was diagnosed in 2 months after the procedure. This case occurred in a hypertensive 63-year-old woman with complex coronary lesions. The coronary angiography showed uncovered proximal dissection with 90% restenosis in left anterior descending coronary artery to left main trunk and 75% restenosis in right coronary artery. We were able to perform semi-emergency off-pump coronary artery bypass (OPCAB). There is the relationship between coronary dissection and restenosis. Cypher stent delayed dissected arterial healing and promote some inflammation at the lesion. Patients implanted Cypher stent with uncovered coronary dissection need more frequent follow-up. PMID- 16715887 TI - [Effects of additional pulmonary blood flow after bidirectional Glenn procedure]. AB - Thirteen cases of functional single ventricle who had undergone bidirectional Glenn procedure were divided into 2 groups according to presence (5) or absence (8) of additional pulmonary blood flow. Additional flow was preserved in cases with relatively small pulmonary artery index (PA index), and their sources were antegrade pulmonary blood flow (2), and Blalock-Taussig (BT) shunt (3). In the control group, PA index was reduced to about 70% of the preoperative value, while in the additional group, pulmonary artery growth was recognized without significant elevation of mean pulmonary artery pressure. However, atrioventricular valve regurgitation progressed and systemic ventricular volume did not decrease after Glenn in the additional group. Therefore special consideration for the timing of Fontan procedure is mandatory. PMID- 16715888 TI - [New method for localization of the small ground-glass opacity lesion in resected lung]. AB - A small lesion showing ground-glass opacity (GGO) by preoperative computed tomography (CT) is sometimes difficult to detect after lobectomy when it locates in the central part of the lobe. In order to facilitate to identify the lesion for marking pathological specimen, we developed a new method using CT. After surgery, the resected pulmonary lobe was expanded with airflow through the bronchial stump and the target lesion was examined with CT. The laser beam of the CT on the surface of the lung is used as a guiding line for cutting. Through the application of this method for 2 clinical cases, it was found to be possible to exactly identify the GGO lesion from the surface of the resected lung enabling to visualize a fresh surface of the lesion like a CT image with minimal destruction of the structure. PMID- 16715889 TI - [Ascending aortic dissection associated with pulmonary contusion caused by blunt trauma]. AB - A 77-year-old female tumbled and struck her front chest wall twice. After the event, hemosputum was observed and she complained of slight chest discomfort. As chest pain continued to worsen a week after the event, she was transferred to our hospital. The computed tomography (CT) revealed an ascending aortic dissection and bilateral pulmonary contusion. In the intensive care unit (ICU), antihypertensive therapy was given immediately and delayed surgical repair was planned because of pulmonary contusion. Twenty days after the event, proximal hemiarch replacement was performed. Postoperative course was uneventful without any complication. She was discharged on the 23rd day after the operation. PMID- 16715890 TI - [Pleomorphic carcinoma of the lung rapidly developed multiple metastases after surgery]. AB - Pleomorphic carcinoma of the lung is a type of carcinoma with spindle and/or giant cells with a poor diagnosis. A 73-year-old male was referred to our hospital because of the pulmonary tumor. Lung biopsy revealed that the tumor was poorly differentiated adenocarcinoma. No distant metastasis were observed by systemic examination. A right middle lobectomy with partial resection of the right upper lobe and lymph node dissection were performed, because the tumor (5.3 x 4.0 x 4.0 cm) was located in peripheral S' and invaded S3 via the interlobular space. Histological findings showed adenocarcinoma comprised of spindle cell components that reacted positively to epithelial membrane antigen (EMA) and no lymph node metastasis. Therefore, he was diagnosed with pleomorphic carcinoma of the lung, pT2N0M0, stage IB. But metastatic lesions newly appeared in the thoracic skin, the liver, the diaphragm, the bilateral adrenal glands, and the retroperitoneal space on the 30th postoperative day. He died of peritonitis and pleuritis on only 60 days after the operation. PMID- 16715891 TI - [Small peripheral lung cancer incidentally detected by massive hemoptysis]. AB - A 47-year-old man was admitted to our hospital for treatment of massive hemoptysis. He was intubated and underwent bronchial arterial embolization because of this deteriorating respiratory state and uncontrollable hemoptysis. Computed tomography (CT) of the chest showed a 1-cm nodular shadow in the peripheral apical lobe (S1) of the right lung. He underwent right upper lobectomy for the purpose of preventing re-hemorrhage as well as making the definitive diagnosis of nodular shadow in S1 of the right lung. Intraoperative pathological examination revealed the nodule as adenocarcinoma, and mediastinal lymphadenectomy was added to the right upper lobectomy. The patient recovered uneventfully, and there has been no sign of recurrence for 15 months after the operation. PMID- 16715892 TI - [Late pseudoaneurysm formation after patch angioplasty of coarctation of the aorta; report of a case]. AB - We report a case of a 13-year-old boy presenting with pseudoaneurysm associated with a knitted Dacron patch used to repair a coarctation of the aorta. At the age of 3 months, he had undergone patch angioplasty for a coarctation of the aorta, which develops following patent ductus arteriosus division at 2 months of age. He was treated by distal aortic arch replacement using 16 mm woven Dacron tube graft in an end-to-end fashion with open proximal anastomosis under deep hypothermic circulatory arrest. The aneurysm was in the aortic wall opposite the patch graft. There was no evidence of infection or dilatation of the patch graft. This case illustrates that repair of aortic coarctations with Dacron patches cannot be recommended. PMID- 16715893 TI - [Aortic regurgitation caused by rupture of a fibrous strand; report of a case]. AB - Rupture of a fibrous strand of a tricuspid aortic valve is extremely rare. We describe a patient with aortic regurgitation due to spontaneous rupture of the fibrous strands that suspend the aortic valve leaflets. The fibrous strands were recognized between the left and right coronary cusps, and between the left coronary and non-coronary cusps during the operation. The fibrous strand between the left coronary and non-coronary cusps had been ruptured, and thus the left coronary and non-coronary cusps were prolapsed into the left ventricle. Since a degenerative change was observed histologically in the resected specimen, these fibrous strands were considered to be an embryonic remnant of the aortic valve. PMID- 16715894 TI - [Elderly patient with acute type A aortic dissection; report of a case]. AB - A 93-year-old woman complained of severe back pain. Computed tomography (CT) and echocardiography revealed acute type A aortic dissection with enlargement of the ascending aorta, moderate aortic regurgitation and cardiac tamponade. Despite her age, the patient was able to support herself, and she and her family desired to have a surgical treatment. Therefore she was referred to our hospital immediately and underwent an emergent ascending aortic replacement using a 28mm woven Dacron graft under deep hypothermic circulatory arrest. Postoperative course was uneventful, and the patient was discharged from our hospital on the 34 th postoperative day in good condition. PMID- 16715896 TI - [Large malignant mesenchymoma of the left posterior mediastinum; report of a case]. AB - Malignant mesenchymoma is a soft tissue tumor arising preferentially in the extremities and retroperitoneum. We report a case of primary malignant mesenchymoma of the left posterior mediastinum. A 24-year-old woman was admitted to our hospital for complaining of cough. Chest X-ray showed a giant mass occupying the 2/3 of the left hemithorax. Chest computed tomography (CT) revealed a lobulated large mass with fat density area and calcified spot in the mediastinum and left pleural space. Chest magnetic resonance imaging (MRI) demonstrated a large solid mass consisting mainly of areas with the same intensity as fatty tissue and partly of areas with heterogenous moderate intensity. The tumor was resected completely under left posterolateral thoracotomy. The pathologic diagnosis was malignant mesenchymoma; well differentiated liposarcoma with osteocartilagenous differentiation. The patient has been well for 3 years and 3 months after surgery. PMID- 16715895 TI - [Mediastinal granular cell tumor: report of a case]. AB - Mediastinal granular cell tumor is rare. We report a case of 16-year-old woman with a granular cell tumor in the right upper-middle mediastinum. Chest computed tomography (CT) and magnetic resonance imaging (MRI) revealed a 4.0 x 2.5 x 5.5 cm well circumscribed mass in the right upper-middle mediastinum. Tumor resection was performed. It was found that the tumor involved right vagus nerve. The tumor was completely excised with combined resection of the right vagus nerve peripheral to the right recurrent nerve. Histopathologically, the tumor consisted of round to polygonal cells with abundant eosinophilic granular cytoplasm, and diagnosed a granular cell tumor. PMID- 16715897 TI - [Positron emission tomography (PET)-computed tomography (CT) suggesting small intestinal metastasis from lung cancer; report of a case]. AB - A 75-year-old man admitted to our hospital due to an abnormal X-ray shadow detected during an annual health check-up. Chest computed tomography (CT) revealed 3.0 cm solid nodules with chest wall invasion in the left lung. We could not get a definitive diagnosis by transbronchial lung biopsy or CT-guided needle biopsy. Positron emission tomography (PET)-CT revealed positive findings in the tumor, aortopulmonary window lymph node and splenic flexure. Under a diagnosis of suspected lung cancer, thoracotomy was performed. As intraoperative diagnosis revealed a moderately differentiated squamous cell carcinoma, the patient underwent a left upper lobectomy, mediastinal lymph node dissection, and combined chest wall resection. Pathological stage was T3N2M0, stage IIIA. Ten days after surgery, the patient suffered from ileus and emergent surgery was performed. Subsequent pathological examination revealed lung cancer metastasis in the small intestine. PMID- 16715898 TI - [Chemical burn]. PMID- 16715899 TI - [Comparison between patient and family report and poison deleterious substance result of analysis]. PMID- 16715900 TI - [Analysis of toxic substances at ER, its present status and usefulness]. PMID- 16715901 TI - [Present status and problems in the management of poisoned patients in ER]. PMID- 16715902 TI - [Drug overdose patients: do all of them need hospitalization?]. PMID- 16715903 TI - [Present condition of the poisoning reason substance analysis in an advanced critical care center]. AB - We investigated the significance of analyzing substances causing intoxication in emergency centers. The poison analysis room was established 6 years before this study. We analyzed a total of 504 patients with intoxication (203 males and 301 females). Concerning the prognosis, 23 patients died; 16 patients (10 males, 6 females) died of paraquat/diquat intoxication, 3 patients (3 males) died of intoxication with organic phosphate agents, 1 patient (male) died of intoxication with a phenoxy agent, and 3 patients (1 male, 2 females) died of intoxication with antipsychotic agents. The other 481 patients were discharged after improvement or referred to another department. Furthermore, we examined the presence or absence of information on intoxication at consultation in the patients with intoxication. In 69 (71.9%) of 96 episodes of intoxication with pesticides, the results of analysis were consistent with the information. In 27episodes (28.1%), there was no information at consultation, or the results of analysis were not consistent with the information. In 299 (79.9%) of 374 episodes of drug poisoning, the results of analysis were consistent with the information. In 75 episodes (20.1%), there was no information at consultation, or the results of analysis were not consistent with the information. These results suggest that the analysis of substances causing intoxication is essential in 20% of patients. We investigated the serum cholinesterase (ChE) level on the initial consultation in the patients with intoxication with pesticides. Ingestion of organic phosphate agents markedly reduced the serum ChE level. However, our results suggest that the serum ChE level depends on the type/amount of pesticides and the interval after ingestion. In addition, many patients took several types of pesticides or drugs, suggesting the importance of analyzing substances causing intoxication. PMID- 16715904 TI - [Case report of repeated acetaminophen overdosing with various symptoms]. AB - A 32-yr-old woman with acute gastritis and migraine used Naron commonly (the principal ingredients are acetaminophen and bromvalerylurea) and had been taking about 3g of acetaminophen daily for several days before admission. She was hospitalized with severe diffuse abdominal pain. On physical examination she had a peritoneal sign and laboratory studies showed elevated liver enzymes, hypophosphatemia, hypokalemia and low blood urea nitrogen(BUN). Serum acetaminophen level was 5.5 microg/mL on admission, so she seemed to be suffered from not only single ingestion but also repeated overdosing. Although we needed for aggressive phosphate and potassium repletion for about a week, all symptoms were distinguished after she quitted acetaminophen ingestion. In addition to hepatic dysfunction, renal failure and disseminated intravascular coagulopathy, we should pay attention to various symptoms like in this case when treating for a acetaminophen poisoning. PMID- 16715906 TI - [Proposal of an analytical pathway for the treatment of poisonings--4--handling of samples, useful information about patient and importance of quantitative analysis]. PMID- 16715905 TI - [Three cases of inhalation of household pyrethroid and metoxadiazone insecticides with remarkable dyspnea]. AB - Household insecticides containing pyrethroids as their active ingredient are widely used in Japan. We encountered three cases of dyspnea caused by insecticides containing pyrethroids and metoxadiazone. The first case was a 52 year-old woman who suffered with dyspnea after breathing interior air following use of a fumigating insecticide. Although bronchoscopy revealed laryngotracheal edema, symptoms improved following administration of hydrocortisone and midazolam. Although the second case was a 63-year-old man who suffered with dyspnea as a result of direct aerosol insecticide inhalation due to missuse, symptoms had dissipated when the patient arrived at the hospital. The third case was a 20-year-old man with allergies who presented with dyspnea due to exposure to an aerosol insecticide three days after use. An allergic mechanism was thought to be involved. Reports of respiratory symptoms and allergic reactions caused by inhalation of pyrethroids are rare. Although the levels of exposure were low in all three cases compared to oral lethal dosage, these cases presented with serious respiratory symptoms, thus suggesting the need for further considerations regarding latent cases and safety. PMID- 16715907 TI - [Organ protection by anesthetics: preface and comments]. AB - Numerous investigations have been performed, focusing on the anesthetic toxicity such as hepatotoxity or nephrotoxicity, for more than 40 years. However, recent basic researche has demonstrated several beneficial effects of anesthetics, including organ protection against ischemia and subsequent reperfusion, and anesthetic preconditioning, as well as clarified mechanisms of acute and delayed cell death, and apoptosis. In this special issue, four experts have provided new relevant information concerning brain, heart, lung, and liver protection by anesthetics, respectively. PMID- 16715908 TI - [Brain protection by anesthetics]. AB - Many investigators have attempted to protect the brain against ischemia by reducing the cerebral metabolic rate using anesthetic agents. However, the magnitude of suppression of the cerebral metabolic rate does not correlate with neuroprotective effects of anesthetics, suggesting that other factors besides reduction in the cerebral metabolic rate contribute to the protection. Facilitation of protein synthesis, GABAergic activity, and anti-oxidant action are likely factors responsible for beneficial effects of barbiturates and propofol. Although the brain is protected during anesthesia, anesthetics cannot provide effects sufficiently enough to recover damage caused by severe ischemia. Further, no desired outcome has been reported by treatments after ischemic events. PMID- 16715909 TI - [Cardioprotection by anesthetics]. AB - Perioperative myocardial ischemia is one of the most important complications associated with significant risk of perioperative cardiac event. Ischemic preconditioning is a phenomenon in which single or multiple brief periods of ischemia have been shown to protect the myocardium against a more prolonged ischemic insult, the result of which is a marked reduction in myocardial infarct size, severity of myocardial stunning, or incidence of cardiac arrhythmias. Myocardial stunning is a clinically important ischemia-reperfusion injury described as a prolonged postischemic contractile dysfunction of myocardium salvaged by reperfusion. Experimental data indicate that general anesthetics protect the myocardium against ischemia-reperfusion injury, as shown by decreased infarct size and a more rapid recovery of contractile function on myocardial stunning. This phenomenon is called anesthetic preconditioning. Volatile anesthetics and morphine have a strong preconditioning like effect. The cardioprotective effect of volatile anesthetics has been supported by some clinical studies. Although the cellular mechanism of anesthetic preconditioning is not fully investigated, possible mechanism involves adenosine, adenosine receptors, the ATP-dependent potassium (K(ATP)) channels, protein kinase C, reactive oxygen species and other mediators or substances. Further, mitochondrial K(ATP) channels play the central role in anesthetic preconditioning. PMID- 16715910 TI - [Lung protection with anesthetics]. AB - Common inhaled and intravenous anesthetics except the barbiturate are recommended for the patients with bronco-constrictive lung disease because of their bronco dilating property. However there are pharmacological potency differences between individual anesthetics. The halogenated inhaled anesthetics and propofol exert antiinflammatory effect on acute lung injury and acute respiratory distress syndrome in the laboratory level, but further study is required for future clinical application. These anesthetics also have an organ protective effect on the ischemia reperfusion lung injury, and their clinical application is expected in the lung transplantation surgery. PMID- 16715911 TI - [Anesthetic protection against hepatic injury]. AB - In this review article, hepatocyte injury by volatile anesthetics, effects of anesthetics on hepatic perfusion, protection offered by either ischemic preconditioning or anesthetic preconditioning against hepatic ischemia reperfusion injury and effects of anesthetics on sepsis-induced hepatic injury are discussed. Halothane poses significant risk of immunologically-mediated hepatocyte injury and disturbances of hepatic blood supply. Other modern volatile anesthetics such as isoflurane, sevoflurane and desflurane seem to have only minor risks. Several animal studies demonstrate that volatile anesthetics offer more protection against ischemia-reperfusion injury than intravenous anesthetics. On the contrary, intravenous anesthetics may be more protective against sepsis induced hepatic injury than volatile anesthetics. PMID- 16715912 TI - [Changes in cardio-pulmonary function during laparoscopic colectomy and postoperative quality of life--comparison with laparotomy]. AB - BACKGROUND: We investigated whether laparoscopic colectomy would affect the degree of respiratory and circulatory changes during surgery and could improve postoperative quality of life (QOL) as compared with laparotomy. METHODS: Fifty one patients (ASA 1-3) scheduled for elective colectomy under general anesthesia with epidural anesthesia were enrolled in the study. All patients were divided into two groups (laparoscopic group: n = 33 and laparotomy group: n = 18) by surgical indication. Respiratory functions and hemodynamic changes were measured during surgery. Postoperative complications, pain scores, the time to start of walking and the period of hospital stay were examined. The serum concentrations of inflammatory cytokines (IL-6, IL-1 ra) and KL-6 were measured perioperatively. RESULTS: It was demonstrated that pneumoperitoneum and postural tilt had a bad influence on respiratory functions especially on pulmonary compliance and a ET(D)co2 in laparoscopic group. The serum concentrations of inflammatory cytokines were significantly lower and the number of rescue for postoperative pain were less in laparoscopic group. The patients in laparoscopic group could begin to walk and leave hospital earlier after surgery. CONCLUSIONS: Severe complication did not occurr in both groups during the perioperative period. Laparoscopic surgery could improve postoperative QOL. PMID- 16715913 TI - [Effects of methylergometrine and oxytocin on blood loss and uterine contraction during cesarean section]. AB - BACKGROUND: The effects of intravenous oxytocics on blood loss and uterine contraction during cesarean section were studied in 136 parturients. METHODS: The subjects were randomized to receive either methylergometrine 0.2 mg bolus (MEM group; n = 34), oxytocin 10 IU over 30 seconds (OX 30 s group; n = 34), oxytocin 10 IU over 5 minutes (OX 5 m group; n = 34) or oxytocin 10 IU over 15 minutes (OX 15 m group; n = 34). The subjects received spinal anesthesia with 11-12 mg of intrathecal isobaric bupivacaine (0.5%). Additional intramyometrial prostaglandin F2alpha (PGF2alpha) was administered when obstetrician diagnosed uterine atony. We analyzed total amount of blood loss including amniotic fluid and number of parturients that received additional intramyometrial PGF2alpha to evaluate uterine contraction. RESULTS: The amounts of blood loss in the OX 30 s and OX 5 m groups were significantly lower than in the MEM group, and the numbers of parturients received additional PGF2alpha in all the oxytocin treat ment groups were significantly lower than in the MEM group (P < 0.05). There were no significant differences in blood loss and uterine contractior among the oxytocin treatment groups. CONCLUSIONS: Intravenous oxytocin 10 IU over 30 seconds to 15 minutes was effective to decrease blood loss and uterine contraction than intravenous methylergometrine 0.2 mg bolus. PMID- 16715914 TI - [Reduction of the concentration of isoflurane prevents tachycardia and hypertension associated with tracheal intubation]. AB - BACKGROUND: High concentration of isoflurane often induces not only tachycardia but also hypertension during induction of anesthesia and causes further hyperdynamic changes after tracheal intubation. METHODS: Forty patients, ASA physical status I, were randomly assigned to receive 4% or 2.5% isoflurane. Anesthesia was induced with thiamylal and vecuronium followed by mask ventilation with 0.5% isoflurane in oxygen. Isoflurane concentration was gradually increased to 4% or 2.5% in 2 min and the trachea was intubated after 3 min. Systolic blood pressure (SBP) and heart rate (HR) were recorded every minute from induction of anesthesia. RESULTS: Mask ventilation with isoflurane induced a significant increase in HR in both groups, but the HR just before intubation was significantly lower in the 2.5% group than in the 4% group. SBP was significantly decreased in the 2.5% group, but a transient increase was seen in the 4% group. Tracheal intubation induced a marked increase in HR in both groups, but the HR was significantly lower in the 2.5% group than in the 4% group (115 +/- 14 and 130 +/- 18 beats x min(-1), respectively; P < 0.01). SBPs just after intubation were 166 +/- 24 and 154 +/- 20 mmHg in the 4% and 2.5% groups, respectively. The difference between the groups was not significant, but the patients in whom the SBP increased more than 180 mmHg were significantly fewer in the 2.5% group than in the 4% group (P < 0.05). CONCLUSIONS: Reduction of the isoflurane concentration from 4% to 2.5% during induction of anesthesia made the circulation stable, and decreased the incidence of excessive tachycardia and hypertension after tracheal intubation. PMID- 16715915 TI - [Anesthetic management of corpus callosotomy with electrophysiological monitoring: a case report]. AB - We report anesthetic management of a 5-year-old girl for corpus callosotomy indicated for the treatment of intractable epilepsy. The procedure mandated intraoperative monitoring of evoked potentials and electrocorticogram. During the first half of the surgery until the corpus callosum was exposed, anesthesia was maintained with continuous infusion of propofol. Motor and somatosensory evoked potentials were monitored and diagnosed as intact throughout the procedure, with no epileptic activity observed in 32-lead electrocorticogram. Then propofol infusion was replaced with the inhalation of sevoflurane, 2.0% in air/oxygen mixture, which induced epileptic spike-and-wave activities, synchronized between the hemispheres, in electrocorticogram. After the completion of corpus callosotomy, we observed interhemispheric desynchronization of epileptic activities indicating successful surgical intervention. The patient emerged from anesthesia uneventfully with no neurological deficits, and thereafter with decreased incidence of generalized epileptic episodes. We suggest that such switch of anesthetic agents between propofol and sevoflurane should be helpful in intraoperative electrophysiological monitoring for ascertaining both functional preservation and successful intervention during epileptic surgery. PMID- 16715916 TI - [Anesthetic management of a patient with hepatocellular carcinoma with tumor thrombosis extending into the right atrium]. AB - We report a patient with hepatocellular carcinoma whose tumor thrombus was extending into the right atrium. A 55-year-old man underwent extended anterior segmental hepatectomy and removal of the tumor thrombus in the right atrium under cardiopulmonary bypass and general anesthesia using sevoflurane, nitrous oxide and oxygen. End-tidal carbon dioxide was closely monitored to detect pulmonary tumor emboli during the surgery. Intravenous fluid and blood transfusion in large volumes were necessary to avoid circulatory derangement. PMID- 16715917 TI - [A case of anaphylactic shock in an elderly man following protamine sulfate administration during emergent off-pump coronary artery bypass grafting]. AB - An 80-year-old diabetic man undergoing emergent off-pump coronary artery bypass grafting for acute myocardial infarction developed anaphylactic shock immediately following administering a small dose of protamine sulfate. Preoperative examination revealed atrial fibrillation, severe three-vessel coronary artery disease and impaired left ventricular function with ejection fraction of 40% and severe septal as well as apical hypokinesis and akinesis. After successful completion of coronary bypass grafting, a total of 40 mg of protamine sulfate was given through the central venous line. Three minutes after protamine administration, profound hypotension occurred. Pulmonary artery pressure was low and the left ventricle was almost empty by transesophageal echocardiography. Hypotension was refractory to rapid administration of 2 l of crystalloid and albumin, and repeated administrations of phenylephrine. Blood pressure finally returned towards baseline after infusion of norepinephrine 0.2 microg x kg(-1) x min(-1) and epinephrine 0.1 microg x kg(-1) x min(-1). Hemoconcentration and impaired oxygenation were also noted. The situation suggested anaphylactic shock due to protamine. He had diabetes mellitus for 20 years and been treated by protamine containing insulin. Postoperative interview revealed that the patient had experienced urticaria over the abdominal area with neutral protamine hagedorn (NPH) insulin administration. This history suggested that the patient had been sensitized by protamine before surgery. Although it is rare to experience anaphylactic shock due to protamine, it is important to elicit the detailed allergic history to insulin in diabetic patients. Because anaphylactic shock still carries high mortality even in a patient without cardiac disease, we were lucky to save this elderly patient with acute myocardial infarction and compromised left ventricular function. PMID- 16715918 TI - [Anesthetic management of pneumonorrhaphy for one-lung patient]. AB - Selective immobilisation of the lobes of the lungs with a bronchial blocker is sometimes a challenge for anesthesiologists. A pneumonorrhaphy was planned in a patient who had had undergone left pneumectomy, and selective immobilisation of the right upper lobe was requested. The initial attempt to place the cuff of a bronchial blocker was unsuccessful. We inserted a urethral guide-wire through fiberoptic bronchoscope and positioned the tip at the orifice of the right upper bronchus. We then threaded the bronchial blocker through the guidewire and placed the cuff at the right upper bronchus. The right upper lobe was successfully collapsed, which greatly facilitated the surgical procedure. The method is easy, fast, and safe and should be considered when difficulty in placement of the bronchial blocker is encountered. PMID- 16715919 TI - [Safe management of anesthesia with total intravenous anesthesia using propofol, fentanyl and ketamine for a patient treated with intraoperative radiotherapy]. AB - A 3-year-old girl with neuroblastoma was scheduled for intraoperative radiation therapy. Prior to surgery, meetings were held to ensure the safety during transfer between the operating room and the radiation unit because those units were placed on the different floors in our hospital. In the operating room the patient was anesthetized with propofol, fentanyl and ketamine (PFK). After the resection of the tumor, the patient was moved to the radiation unit. All personnel had to leave the treatment room while the radiation was administered, which required 3 minutes. During the transfer and treatment, the following apparatus were effective to ensure the patient's safety; mobile respiratory and cardiovascular monitors, closed-circuit television to observe the monitors and patient, and a portable gas powered ventilator. The total intravenous anesthesia with PFK was used throughout all scheduled procedures. For patients treated with intraoperative radiation therapy, the administration of inhalation anesthetics should be discontinued to supply 100% oxygen, which enhances the effect of irradiation, and to avoid the complexity of the transfer with an anesthetic machine. This discontinuation, however, may cause the cardiovascular instability. It seems that total intravenous anesthesia has advantages over inhalation anesthesia for patients who undergo intraoperative radiation therapy. PMID- 16715920 TI - [Successful resuscitation of intractable hyperkalemic cardiac arrest]. AB - We report on a 42-year-old oliguric uremic man on regular hemodialysis who developed sudden cardiac arrest, secondary to severe hyperkalemia, with a plasma potassium concentration of 9.7 mEq x l(-1). The cardiac arrest persisted after the initiation of cardiopulmonary resuscitation and intensive treatment for marked hyperkalemia for an hour and 55 minutes. Therefore a portable percutaneous cardiopulmonary support (PCPS) system had to be instituted while the patient had very prolonged refractory ventricular fibrillation. His cardiac rhythm was restored immediately after application of PCPS and he recovered without neurological sequelae. We therefore suggest that PCPS should be considered as a therapeutic option during cardiopulmonary resuscitation for life-threatening cardiac arrest secondary to severe hyperkalemia. PMID- 16715921 TI - [Postoperative acute pulmonary thromboembolism treated with nafamostat mesilate]. AB - A 66-year-old woman underwent total gastrectomy. On the first postoperative day, acute pulmonary thromboembolism (PTE) with occlusion of the pulmonary trunk occurred, and the cardiovascular state became unstable. We used urokinase and heparin at lower doses. However, massive bleeding as a result of these drugs made it difficult to continue thrombolytic and anticoagulative therapies. We stopped urokinase on the second postoperative day, and started nafamostat mesilate instead of heparin on the third postoperative day. We found that the targeted activated coagulation time value (180 sec) was more easily attained by nafamostat mesilate than by heparin. We could continue anticoagulation therapy with minimal bleeding thereafter, and the patient recovered smoothly. On CT of the 19 th postoperative day, a small thrombus was observed in the right pulmonary artery. PMID- 16715922 TI - [Leak of anesthetic gases inside the anesthesia machine]. AB - We experienced the leak of anesthetic gases inside the anesthesia machine in spite of performing the leak test before its use. After induction of anesthesia, a laryngeal mask airway was inserted and the patient was ventilated manually. At the beginning we could not find any signs of machine troubles. High airway pressure occurred immediately after switching to the mechanical ventilation. Because we could not detect the details of the machine trouble, tidal volume was set lower and the surgery was continued. After surgery, we found a crack in a fresh gas circuit valve. We have to check the anesthesia machine regularly and know its duration of use. PMID- 16715923 TI - [Tension pneumothorax associated with re-intubation through percutaneous tracheostoma site]. AB - Percutaneous tracheostomy (PT) has become popular recently, but occasionally cannula exchange can be difficult and hazardous. A 55-year-old woman with pontine hemorrhage was admitted to our ICU after oral intubation. On the 3 rd day, PT was performed with no complication to prepare for prolonged airway management. On the 14 th day, surgical removal of the pontine hematoma was scheduled. Since our neurosurgeon requested a prone position with maximal neck anteflexion, she was re intubated orally and the tracheostomy cannula was removed during the operation. At the end of the operation, the tracheostoma had already been narrowed and re insertion of a narrower tracheostomy cannula was unsuccessful even with bronchoscopic guidance. Following several attempts, her Spo2 gradually decreased and her blood pressure dropped. A chest X-ray showed right pneumothorax and a chest drainage tube was inserted. Posterior tracheal wall injury was suspected, though the exact injury site was not identified bronchoscopically. She was treated conservatively because no air leak was observed after advancing the oral tube. Three weeks later, surgical tracheostomy was performed without major abnormal findings. A small tracheostoma characteristic of PT might be associated with an increased risk of delayed airway complication. Cannula exchange should be performed more carefully after PT compared with surgical tracheostomy. PMID- 16715924 TI - [In vitro response time of continuous cardiac output measurement in STAT mode of Vigilance after controlled flow change]. AB - BACKGROUND: Continuous cardiac output measurement in STAT mode (STAT CCO) equipped with Vigilance displays cardiac output every 30 to 60 seconds. The aim of this study is to verify the hypothesis that each value with this system is computed only from the data collected in one update period. METHODS: The circuit was filled with normal saline and flowed by a roller pump in in vitro setting. The flow rate was set at either 2.5 l x min(-1) or 5.0 l x min(-1) and changed quickly to another state after each state had been maintained for 25 minutes. The change operation was repeated 10 times. The maximum difference was defined as the difference between the value at the start and the maximum change value. The response time was defined as the time from the start to the time to reach 80% of the maximum difference. In each operation, the response time of STAT CCO was calculated. RESULTS: The response time of STAT CCO was 9.7 +/- 1.3 min (mean +/- SD). CONCLUSIONS: The response time of STAT CCO was about 10 times longer than one update period. This result suggests that STAT CCO values are not computed only from the data collected in one update period. PMID- 16715925 TI - [Yale anesthesiology: a reflection on Professor Kitahata's days]. AB - Professor Luke Masahiko Kitahata was a professor of anesthesiology, Yale University in the United States of America. He had been employed in Yale University for 33 years, from 1964 to 1997, and acted as the chairman of the department of anesthesiology from 1973 to 1982. He endeavored not only to achieve his academic career but also to educate his juniors. The number of Japanese fellows who were trained by Professor Kitahata is more than sixty. He rendered many services for the development of anesthesiology in Japan. PMID- 16715926 TI - [The problems in anesthetic support in developing countries]. AB - We report our support for anesthesia in Afghanistan and northern Iraq. The anesthetic staff of both countries expressed their wish to learn regional anesthesia. In Afghanistan, anesthesia is carried out only by nurse anesthetists, who need basic education in anesthesia. Due to the shortage of medical supply, the present condition is not suitable for regional anesthesia training. In northern Iraq, there are adequate medical supply and competent anesthetists, and the training of regional anesthesia was possible. The sustainability of health care depends on its country's economic power. The technical support as an aid is difficult, if the level of support exceeds the capacity of the economic power. PMID- 16715927 TI - [Central nervous system can regenerate! Raising expectations for therapeutic application for neurologic disorders in children]. PMID- 16715928 TI - [An 11-year-old girl showing hypotonia and dying of renal failure]. PMID- 16715929 TI - [Woman physicians in child neurology--their opinions and dreams (discussion)]. PMID- 16715930 TI - [Study of situational recognition of attention deficit/hyperactivity disorders, Asperger's disorder and high functioning autism with the Metaphor and Sarcasm Scenario Test (MSST)]. AB - The purpose of this study is to clear up the difference between Asperger's disorder (AS) and high functioning autism (HFA) with a newly developed test, the metaphor and sarcasm scenario test (MSST) can be used to distinguish pervasive developmental disorder (PDD) from attention deficit/hyperactivity disorders (AD/HD). So 66 AS children, 20 HFA children and 37 AD/HD children were selected, compared the profiles of score on the MSST. The results showed that the inability to understand sarcastic situation was specific to group with AS, both group with HFA and AD/HD were differ little to understand between metaphor and sarcasm. In this study suggest differences in situational recognition among the PDD subtypes, the clinical symptoms of PDD reflects characteristics of AS. PMID- 16715931 TI - [Study of the kid-kINDL questionnaire scores for children with developmental disorders in normal classes and their parents]. AB - The KINDL-questionnaire is able to measure degrees of health and adaptability in relation to quality of life (QOL) in children. The questionnaire can be completed by children, adolescents, and their parents (parent-version). We had translated Kid-KINDL questionnaire into Japanese as Questionnaire for Measuring QOL in Japanese Elementary School Children, and reported their reliability and validity. This study investigates Japanese elementary school version of the Kid-KINDL questionnaire scores consisting of 6 dimensions (4 items each;total score, 100) for children with developmental disorders without mental retardation including high-functioned pervasive developmental disorder, attention deficit/hyperactivity disorder and learning disorder, and the parent-version scores for their mothers. Twenty individuals in normal class and their mothers participated after informed consents were obtained. Their total QOL scores were significantly lower than those in control group. Four of 6 dimensions consisting of emotional well-being, self-esteem, family, friends and school had significantly lower points. Their mothers' points in total QOL scores and all dimensions except for family dimension were significantly lower than those in control group. In comparison of scores between children with mild developmental disorders and their mothers, children estimated lower in self-esteem and family dimensions instead mother estimated lower in health, emotional well-being and school dimensions. There was no difference in total scores between them. Children with mild developmental disorders and their mothers estimate lower points of QOL scores than those of the normal control group. There are different perceptions in 5 of 6 dimensions between the children and their mothers. PMID- 16715932 TI - [Development of auditory-visual spatial integration using saccadic response time as the index]. AB - We measured saccadic response time (SRT) to investigate developmental changes related to spatially aligned or misaligned auditory and visual stimuli responses. We exposed 4-, 5-, and 11-month-old infants to ipsilateral or contralateral auditory-visual stimuli and monitored their eye movements using an electro oculographic (EOG) system. The SRT analyses revealed four main results. First, saccades were triggered by visual stimuli but not always triggered by auditory stimuli. Second, SRTs became shorter as the children grew older. Third, SRTs for the ipsilateral and visual-only conditions were the same in all infants. Fourth, SRTs for the contralateral condition were longer than for the ipsilateral and visual-only conditions in 11-month-old infants but were the same for all three conditions in 4- and 5-month-old infants. These findings suggest that infants acquire the function of auditory-visual spatial integration underlying saccadic eye movement between the ages of 5 and 11 months. The dependency of SRTs on the spatial configuration of auditory and visual stimuli can be explained by cortical control of the superior colliculus. Our finding of no differences in SRTs between the ipsilateral and visual-only conditions suggests that there are multiple pathways for controlling the superior colliculus and that these pathways have different developmental time courses. PMID- 16715934 TI - [Intelligence of hemiplegic children: an association with epilepsy and the laterality of the lesion]. AB - We studied thirty-one children with hemiplegia to elucidate the relationship of intelligence, the laterality of a lesion, and epilepsy, in these children. We investigated the influences of the age at the onset of the seizures, the number of anti-convulsants, and the duration of medication, on the DQ/FIQ. The results indicated a significantly higher DQ/FIQ and VIQ for right hemiplegic children than for left hemiplegic children. But the PIQ of the patients with either a left or right hemiplegia was not significantly different. A significantly higher DQ/FIQ, VIQ, and PIQ was also revealed for the children without epilepsy. In addition, there was also a tendency that the DQ/FIQ of children who suffered from epilepsy before the age of one year was lower than that of children who suffered from epilepsy after the age of one year. The present results provide evidence which supports the crowding hypothesis. PMID- 16715933 TI - [Neurological complications during and after the treatment of acute lymphocytic leukemia]. AB - To explore the frequency and prognosis of neurological complications of acute lymphocytic leukemia, retrospective studies were made of patients with acute lymphocytic leukemia. Neurological complications were found in 13 of 100 patients during and after treatment. They were caused by chemotherapy in 8 patients, irradiation therapy in 2, vitamin B1 deficiency in 1, and unknown in 2. Medications primarily relevant to these complications were methotrexate in 5 patients, L-asparaginase in 2, cytosin arabinoside in 1. The patients were diagnosed as having leukoencephalopathy (8), cerebrovascular injury (4), and Wernicke's encephalopathy (1). Symptomatic epilepsy was found in one patient, and mental retardation was seen in three patients during a 2-year course of treatment. We conclude that careful management is required in the treatment of acute lymphocytic leukemia, because of the persistence of neurological complications, although their severity is decreasing with advances in treatment. PMID- 16715936 TI - [Secondary progressive multiple sclerosis in childhood--interferon beta 1b treatment]. AB - Multiple sclerosis (MS) is rare in children and the efficacy and safety of interferon beta 1b (IFN-beta 1b) treatment in childhood MS has not yet been established. We started to treat a boy who suffered from relapsing remitting MS with IFN-beta 1b at 8.5 years of age, because he had severe neurological disability in consequence of frequent relapses and incomplete remission. After initiating IFN-beta 1b treatment, his clinical course moved to secondary progressive (SP) MS, and he demonstrated poor improvement in degenerative progression and his disability continued to worsen. We could speculate that IFN beta 1b was not effective regarding the degenerative component of childhood MS as in that of adult MS. SPMS is extremely rare in children, and this case would provide a very important implication to predict the tolerability of IFN-beta 1b treatment depending on the type of clinical course in childhood MS. PMID- 16715935 TI - [Polycystic ovary syndrome and hepatocellular adenoma related to long-term use of sodium valproate in a young woman]. AB - Previous studies have reported a high prevalence of polycystic ovary syndrome (PCOS) among women taking sodium valproate (VPA). We report the case of a 28 year old epileptic female taking VPA, who developed PCOS and later hepatocellular adenoma. She had been taking VPA for intractable epilepsy since the age of 15 months. At the age of 22 years, she suffered spontaneous rupture of a liver tumor that was diagnosed as hepatocellular adenoma. At the age of 24 years, bilateral polycystic ovaries were found by transabdominal ultrasonography, and PCOS was diagnosed. VPA may directly influence steroidogenesis in the ovary and cause hyperandrogenemia with ensuing PCOS. It is known that abnormality in the sex hormones contributes to the onset of hepatocellular adenoma. Therefore, we speculate that hyperandrogenemia due to VPA contributed to the development of hepatocellular adenoma in this case. PMID- 16715937 TI - [Changes of electroencephalographic findings in a case of migraine with various neurological symptoms]. AB - We report here an 1-year-old boy who was diagnosed as sporadic hemiplegic migraine demonstrated transient changes in electroencephalographic (EEG) background activity accompanied with migraine attacks. The attacks were associated with various neurological complications such as impairment of consciousness and hemiplegia and so on, which coincided with the changes of EEG findings. When his first headache developed with fever and impairment of consciousness, diffuse slow waves (1-2 Hz, 250-300 microV) appeared transiently. The slow wave gradually improved thereafter, and disappeared on the 24th day. On the 25th day, when he had the second attack with left hemiplegia and impairment of consciousness and sensation, slow waves, (2-3 Hz, 250-300 microV) appeared on right side of the brain. Constriction of the right middle cerebral artery was found on MR angiography at that time. The EEG change had improved to normal background activity 3 days after the episode. This unique case demonstrated that transient EEG changes seem to be correlated with the existence of unconsciousness and the laterality of the ischemic insult. We may assume that the changes of electroencephalographic findings may represent the cortical dysfunction caused by vascular constriction. PMID- 16715938 TI - [Case of theophylline-induced seizures successfully treated with mild hypothermic therapy and methylpredonisolone pulse therapy]. PMID- 16715939 TI - [Smooth introduction of nasal intermittent positive pressure ventilation (NIPPV) for a patient with Fukuyama type congenital muscular dystrophy with severe mental retardation]. PMID- 16715940 TI - [Molecular epidemiology of tuberculosis by the use of IS6110 restriction fragment length polymorphism: a study from 2001 to 2003]. AB - OBJECTIVE: To analyze the situation of tuberculosis infection by DNA fingerprinting in the middle and eastern part of Osaka, Japan. DESIGN: We performed IS6110 restriction fragment length polymorphism (RFLP) on 1200 isolates from tuberculosis patients who visited our hospital from January 2001 to December 2003. A cluster was defined as a series of isolates with more than 90% similarity by IS6110 RFLP and those with the same drug-susceptibility pattern. The isolates with fewer than six copies of IS6110 were considered to be clustered if the IS6110 RFLP patterns and the variable numbers of tandem repeats with 16 regions of ETR and MIRU "allele profile" were identical. RESULTS: The number of samples in incremental study periods was 422 in 2001, 817 between 2001 and 2002 and 1200 between 2001 and 2003. The percentage of clustered cases was 27.8% in 2001, 19.1% in 2002 and 19.5% in 2003. The cumulative percentage of clustered cases was 27.8% in the first year, 29.7% over two years and 32.6% over three years. The percentage of clustered cases of isolates with a drug resistance was significantly lower (25.0%) than that of drug susceptible isolates (33.7%). Next, we investigated the clustered cases by gender and age. The percentage of clustered cases with isolates from young males and females (0-19 years old) was 23.8%. In contrast, the percentage of clustered cases with isolates from 20-59 year-old females gradually decreased from 14.7% to 4.4%. Conversely, the percentage of clustered cases from young and middle aged male (20-59 years old) was higher (20.2%-32.4%) than that of females. CONCLUSION: The sharp increase in the cumulative cluster formation rate was curbed by the decline in the tuberculosis incidence rate in Osaka, Japan, after the first year of examination. We thought that this phenomenon suggests the success of the anti-tuberculosis measure in Japan. PMID- 16715941 TI - [Antimycobacterial susceptibility against nontuberculous mycobacteria using brothmic NTM]. AB - PURPOSE: Recently the incidence of pulmonary nontuberculous mycobacteria infection has increased among patients not only implicated with AIDS, but also without predisposing conditions. However, an effective antimicrobial therapy for the disease has not been established yet, because of the absence of highly active therapeutic drugs. We compared the in vitro antimicrobial activities of five antituberculous drugs, clarithromycin and fluoroquinolones against 92 clinical isolates belonging to three species of slowly growing nontuberculous mycobacteria. MATERIAL AND METHODS: M. avium (31 strains), M. intracellulare (44 strains), and M. kansasii (17 strains), all of which were isolated from sputum specimens of previously untreated patients with pulmonary nontuberculous mycobacteria infection, were used. The eight agents tested were streptomycin, ethambutol, kanamycin, isoniazid, rifampicin, clarithromycin, levofloxacin and gatifloxacin. The drug susceptibility of these strains in terms of MIC (minimum inhibitory concentration) was determined by BrothMIC NTM. RESULTS: The MICs of rifampicin, clarithromycin, levofloxacin and gatifloxacin for all three species were low and gatifloxacin was more active than levofloxacin between two fluoroquinolones. Regarding clarithromycin, 100% of the strains were susceptible to 2 micrograms/ml or less and none of the strains were resistant on this level. In contrast, the MICs of ethambutol and isoniazid for M. avium and M. intracellulare were high and less active in vitro than the other antimicrobial agents. CONCLUSION: These MIC studies suggest that rifampicin, clarithromycin, levofloxacin, and gatifloxacin have excellent in vitro antimicrobial activities against M. avium, M. intracellulare and M. kansasii and especially clarithromycin may be very useful as a drug therapy for previously untreated patients. In the treatment of pulmonary nontuberculous mycobacterium infection, further studies are needed to evaluate the clinical effects of these drugs and to observe the drug resistance, on the basis of the results of the drug susceptibility test by BrothMIC NTM. PMID- 16715942 TI - [In vitro antituberculous activity of ofloxacin and levofloxacin against multidrug-resistant tuberculosis and clinical outcomes]. AB - OBJECTIVE: To investigate in vitro antituberculous activity of ofloxacin (OFLX) and levofloxacin (LVFX) against multidrug-resistant tuberculosis and to study the clinical outcomes. SUBJECTS AND METHODS: In vitro antituberculous activity of OFLX and LVFX against multidrug-resistant strains of Mycobacterium tuberculosis isolated from 46 patients with pulmonary tuberculosis and a retropective clinical analysis of 45 patients were investigated. RESULTS: In susceptibility testing, resistance rates to OFLX or LVFX were higher in intractable cases (7/20: 35%) and in cases with prior chemotherapy using new quinolones (5/12: 42 %). Sputum culture conversion was observed in 34 patients (76%), however 9 among them later reverted to positive culture. In a single variate proportional hazards model, risk factors related to poor outcomes (treatment failure or relapse) were resistance to OFLX or LVFX, advanced disease on chest radiograph, and the number of susceptible drugs four or less. In a multiple variate proportional hazards model, a risk factor was resistance to OFLX or LVFX. Eighteen patients (40%) died, and among them, 10 died of tuberculosis. Survival time of treatment failure patients was significantly shorter than patients with sputum culture conversion. CONCLUSION: Resistance to OFLX or LVFX was considered to be a risk factor related to treatment failure and relapse in multidrug-resistant tuberculosis. PMID- 16715943 TI - [A case of severe intestinal tuberculosis, treated with ciprofloxacin, kanamycin and prednisolone]. AB - A 31-year-old man was admitted to our hospital because of frequent diarrhea. Colonoscopy showed ring ulcers on the rectum and ascending colon and chest X-ray showed abnormal shadows which were diagnosed as tuberculosis by sputum PCR. He started treatment with isoniazid (INH), rifampicin (RFP), pyrazinamide (PZA) and streptomycin (SM), however, eruption and ileus were seen. Then, he was retreated with ciprofloxacin (CPFX), kanamycin sulfate (KM) and prednisolone (PSL). Subsequently, we added RFP and further added calcium para-aminosalicylate (PAS). All these treatment was effective, and he was discharged from the hospital. PMID- 16715944 TI - [A case of pulmonary tuberculosis complicated with an orthotopic liver transplantation]. AB - The infectious disease is one of the most important complications related to the organ transplantation. Patients using immunosuppressive agents often present atypical tuberculosis and the treatment of such case is far more difficult in some cases due to the liver damage and/or the drug interaction. We report a case of pulmonary tuberculosis in a patient of 60-year-old man using tacrolimus after an orthotopic liver transplantation. He had liver transplanted orthotopically for the long-term history of chronic hepatitis B and subsequent liver failure on January 28, 2004. An abnormal shadow was first detected on his chest X-ray film on October, 2004. He was admitted to our hospital after the smear of the gastric juice showed some acid-fast bacilli and tubercle bacilli were confirmed by polymerase chain reaction (PCR). Tuberculin skin test was positive (erythema 10 x 10) and the computed tomography (CT) scan of his chest revealed a nodular opacity with some smaller nodules scattered around in the right upper lobe. We started four anti-tuberculous drugs other than pyrazinamide (PZA) and rifampicin (RFP), which included isoniazid (INH), ethambutol (EB), streptomycin (SM), levofloxacin (LVFX). The liver enzyme was transiently elevated (AST 123 IU/I, ALT 103 IU/I) but improved after desensitization against INH. The blood concentration of tacrolimus preserved between 5 and 7 ng/ml and there was no need to change the dosage. PMID- 16715945 TI - In vivo bioluminescence imaging of cord blood derived mesenchymal stem cell transplantation into rat myocardium. AB - OBJECTIVE: The conventional method for the analysis of myocardial cell transplantation depends on postmortem histology. Here, we have sought to demonstrate the feasibility of a longitudinal monitoring of transplanted cell survival in living animals, accomplished with optical imaging techniques and pharmacological interventions. METHODS: Human cord blood (50 ml) was donated with parental consent. After getting cord blood derived mesenchymal stem cells (CBMSCs), cells were transfected (MOI = 100) overnight with adenovirus encoding firefly luciferase gene (Ad-CMV-Fluc). Our experimental Sprague-Dawley rats (n = 12) were given intramyocardial injections containing 1 x 10(6) CBMSCs, which had been made to express the firefly luciferase (Fluc) reporter gene. Optical bioluminescence imaging was then conducted using a cooled charged-coupled device (CCD) camera (Xenogen), beginning on the day after the transplantation (day 1). Groups of mice were intraperitoneally injected with cyclosporine (5 mg/kg) or tacrolimus (1 mg/kg), in an attempt to determine the degree to which cell survival had been prolonged, and these values were then compared with the cell survival values of the negative control group. The presence of transplanted CBMSCs on in vivo images confirmed by in situ hybridization for human specific Alu in the myocardium. RESULTS: Cardiac bioluminescence signals were determined to be present for 6 days after transplantation: day 1 (97000 +/- 9100 x 10(5) p/s/cm2/sr), day 3 (9600 +/- 1110 p/s/cm2/sr), and day 5 (3200 +/- 550 p/s/cm2/sr). The six mice that received either cyclosporine or tacrolimus displayed cardiac bioluminescence signals for a period of 8 days after transplantation. We observed significant differences between the treated group and the non-treated group, beginning on day 3 (tacrolimus; 26500 +/- 4340 p/s/cm2/sr, cyclosporine; 27200 +/- 3340 p/s/cm2/sr, non-treated; 9630 +/- 1180 p/s/cm2/sr, p < 0.01), and persisting until day 7 (tacrolimus; 12500 +/- 2946 p/s/cm2/sr, cyclosporine; 7310 +/- 1258 p/s/cm2/sr, non-treated; 2460 +/- 160 p/s/cm2/sr, p < 0.01). The human-derived CBMSCs were detected in the myocardium 3 days after transplantation by in situ hybridization. CONCLUSIONS: The locations, magnitude, and survival duration of the CBMSCs were noninvasively monitored with a bioluminescence optical imaging system. We determined that optical molecular imaging expedites the fast throughput screening of pharmaceutical agents, allowing for the noninvasive tracking of cell survival within animals. In rat cardiac CBMSC transplant models, transient immunosuppressive treatment with tacrolimus or cyclosporine was shown to improve donor cell survival. PMID- 16715946 TI - Prognostic value of FDG-PET in patients with ovarian carcinoma following surgical treatment. AB - OBJECTIVE: [corrected] To determine the prognostic value of FDG-PET after surgical resection in patients with ovarian carcinoma, we compared the results of FDG-PET and serum CA-125 level and prognosis of patients. METHODS: Eighteen patients underwent a total of 32 FDG-PET examinations following surgery for ovarian carcinoma from October 2001 to December 2002 at our hospital (median follow-up period, 31 months). Age of the patients at the time of the initial FDG PET examination ranged from 31 to 73 years (mean 52 years) and the period from surgery to the initial FDG-PET examination ranged from 5 to 109 months (mean 30 months). Serum CA-125 levels were determined on the occasion of each FDG-PET examination. Recurrent tumors were treated with surgery in 5 cases, radiotherapy in 2 cases, and chemotherapy in 9 cases. RESULTS: The initial FDG-PET examinations revealed that 13 cases had positive and 5 cases had negative findings, which included 2 false positive cases. The survival rate for all patients at 1 year and 2 years after the initial examination was 82% and 63%, respectively. Two-year survival rates in patients with positive and negative FDG PET findings were 51% and 83%, respectively, and the difference was not statistically significant (p = 0.19). Furthermore, 4 patients with normal CA-125 levels and 14 patients with elevated CA-125 levels showed 2-year survival rates of 100% and 51%, respectively, and they were not significantly different (p = 0.11). For all 32 examinations, the 2-year survival rates for patients with normal CA-125 levels (100%) were significantly higher (p = 0.025) than that for patients with elevated CA-125 levels (47%), however there was no significant difference (p = 0.20) between FDG-PET positive cases (53%) and negative cases (83%). CONCLUSION The prognosis of patients with positive FDG-PET findings was less favorable than that of patients with negative findings. However, over the mean extended observation period of about 2.5 years, no significant difference in the prognosis of patients was observed between the two groups. The results of the present study indicate that elevated serum CA-125 levels may be more useful for evaluating the prognosis of ovarian cancer during the post-operative follow-up than FDG-PET findings. PMID- 16715948 TI - Comparison of extraarticular leakage values of radiopharmaceuticals used for radionuclide synovectomy. AB - OBJECTIVES: Radionuclide synovectomy is a reliable therapy in patients with chronic synovitis. However, radiation doses delivered to non-target organ systems due to leakage of radioactive material from the articular cavity are an important disadvantage of this procedure. In this study we compared extraarticular leakage values of the 3 commonly used radiopharmaceuticals; 90Y-citrate, 90Y-silicate and 186Re-sulfide colloid. MATERIALS AND METHODS: Thirty-five patients with persistent synovitis were enrolled in the study. Twenty-two hemophilic, 8 rheumatoid arthritis and 5 patients with pigmented villonodular synovitis were studied. 90Y labeled silicate and citrate were used for knee joints and 186Re sulfide for intermediate sized joints. Radiocolloid leakage values were evaluated using a gamma camera with 20% window centered over the bremsstrahlung photopeak of 90Y and a respective window over the 137 keV photopeak of 186Re. Regions of interest were drawn over the injection site, the regional lymph nodes and the background areas. Leakage of radiocolloid was calculated by dividing the counts/pixel in the regional lymph node area to the counts/pixel in the injection site. RESULTS: No visible leakage was observed. The median leakage values calculated for 90Y-citrate, 90Y-silicate and 186Re-sulfide were found as 1.9%, 2.4% and 2.7%, respectively. The difference between the variability of leakage values was not statistically significant (p > 0.05). CONCLUSION: There was no significant difference in terms of extraarticular leakage between 9Y-citrate, 9Y silicate and 186Re-sulfide radiocolloids. PMID- 16715947 TI - Detection of maleate-induced Fanconi syndrome by decreasing accumulation of 125I 3-iodo-alpha-methyl-L-tyrosine in the proximal tubule segment-1 region of renal cortex in mice: a trial of separate evaluation of reabsorption. AB - OBJECTIVE: Fanconi syndrome is a renal dysfunction characterized by various combinations of renal tubular transport dysfunction involving amino acids, glucose, protein and other substances. Most reabsorption of amino acids occurs in proximal renal tubule segment 1 (S1). The present study evaluated the possibility of early detection of drug-induced Fanconi syndrome, based on decreased renal accumulation of 125I-3-iodo-alpha-methyl-L-tyrosine (125I-IMT), an amino acid transport marker, in the S1 region of renal cortex. The present experimental model used maleate (MAL)-induced Fanconi syndrome in mice. Results were compared between 125I-IMT and 3 other clinical renal radiopharmaceuticals: 99mTc-2,3 dimercaptosuccinic acid (99mTc-DMSA); 99mTc-mercaptoacetylglycylglycylglycine (99mTc-MAG3); and 99mTc-diethylenetriaminepentaacetic acid (99mTc-DTPA). METHODS: Male ddY mice (age, 6 weeks; body weight, 25 g) were used to create a Fanconi model of renal dysfunction. A single dose of maleate disodium salt was administered by intraperitoneal injection (6 mmol/kg). Hematoxylin and eosin (HE) staining of the renal cortex, renal autoradiography and measurement of renal radioactivity of labeled compounds were performed at 30, 60, 90 and 120 min after MAL injection. At 5 min after injection of labeled compounds (18.5 kBq for accumulation experiment, 670 kBq for autoradiography), animals were sacrificed by ether overdose and kidneys were removed. For the accumulation experiment, radioactivity was measured using a well-type scintillation counter. For autoradiography, 20-microm sections of frozen kidney were used with Bio-Imaging Analyzer. RESULTS: At 30 min after MAL injection, HE staining showed pyknosis in some proximal tubule cells. At that time, accumulations of 125I-IMT and 99mTc DMSA in the S1 region were approximately 67% and 55% of control levels (p < 0.005). MAL increased accumulation of 99mTc-DTPA in the S1 region, but had no effect on accumulation of 99mTc-MAG3 in the S 1 region. CONCLUSIONS: Decreased accumulation of 123I-IMT in the S1 region appears to represent a useful marker for detection of MAL-induced Fanconi syndrome. In future, we plan to assess the efficacy of using 125I-IMT to monitor renal dysfunction induced by nephrotoxic clinical drugs. PMID- 16715949 TI - Performance of list mode data acquisition with ECAT EXACT HR and ECAT EXACT HR+ positron emission scanners. AB - Recently, list mode (event-by-event) data acquisition with positron emission tomography (PET) has been widely noticed because list mode acquisition is superior to conventional frame mode data acquisition in terms of (1) higher efficiency of data storage, (2) higher temporal resolution, and (3) higher flexibility of data manipulation. The aim of this study is to investigate the performance of list mode data acquisition with ECAT EXACT HR and HR+ PET scanners (CTI PET Systems) and its feasibility in clinical applications. A cylindrical phantom (16 cm in diameter and length) filled with a 11C solution for the HR and a 15O solution for the HR+ was scanned several times by varying the radioactivity concentration with the list mode and frame mode acquisitions. The scans were also carried out with a septa (2D mode) and without a septa (3D mode) in order to evaluate the effect of the interplane septa on the quality of the list mode data. The acquired list mode data were sorted into a sinogram and reconstructed using a filtered back-projection algorithm. The count rate performance of the list mode data was comparable to that of the frame mode data. However, the list mode acquisition could not be performed when the radioactivity concentration in the field-of-view was high (exceeding 24 kBq/ml for the 3D mode) due to a lack of sufficient transfer speed for sending data from the memory to hard disk. In order to estimate the pixel noise in a reconstructed image, ten replicated data sets were generated from one list mode data. The reconstructed images with the 3D mode had a signal-to-noise ratio that was more than 60% better than that of the image with the 2D mode. The file size of the generated list mode data was also evaluated. In the case of ECAT EXACT HR+ with the 3D list mode, the list mode data with a generated file size of 2.31 Mbytes/ s were generated for 37 MBq injections. Our results suggest that careful attention must be paid to the protocol of the list mode data acquisition in order to obtain the highest performance of the PET scanner. PMID- 16715950 TI - Surveillance study for creating the national clinical database related to ECG gated myocardial perfusion SPECT of ischemic heart disease: J-ACCESS study design. AB - BACKGROUND: ECG-gated myocardial perfusion SPECT is widely applied to diagnose ischemic heart disease, and such findings are useful to predict patient prognosis. However, Japan does not have a database that correlates SPECT image findings with the prognosis of patients who have ischemic heart disease. METHODS: A large-scale clinical study involving 117 medical facilities throughout Japan was established to survey the clinical background and image findings of patients who have undergone ECG-gated stress perfusion SPECT. These patients were followed up for three years to investigate the occurrence of cardiac events. RESULTS: The 4,629 registered patients comprised 2,989 males (age 64.9 +/- 10.3 y, mean +/- SD) and 1,640 females (age 67.2 +/- 9.7 y). The most frequent complication was hypertension (54.5%), followed by hyperlipidemia (47.2%) and diabetes (29.4%). Percutaneous coronary intervention (PCI) or coronary artery bypass grafting (CABG) was conducted on 1,925 of the patients. SPECT examinations were ordered for further examination of chest pain (32.8%), periodic follow-up after coronary artery intervention (24.2%), screening for coronary artery disease (15.1%), follow-up of old myocardial infarction (14.9%), more detailed investigation of ECG or echocardiographic abnormalities (13.1%), etiological assessment of heart failure (1.6%), and further inspection for acute coronary syndrome (0.3%). The method of inducing stress was most often exercise loading at 68.8%, and infusion of either dipyridamole (14.6%) or adenosine triphosphate (ATP, 13.8%). The most frequently applied amount of 99mTc-tetrofosmin was an initial dose of 200 to 300 MBq combined with a second dose of 700 to 800 MBq (37.7%). The mean doses were 305 +/- 81 at the initial and 709 +/- 132 MBq at the second administration. A history of angina pectoris (41.2%) was the most frequent, followed by myocardial infarction (29.5%). CONCLUSIONS: During the two years of follow-up after registration, 46 of the 4,629 subjects have discontinued or dropped out, 134 have died, and 4,449 (97.8%) continue to undergo follow-up investigations. A complete report will be presented when the follow-up data for 3 years have been compiled and analyzed. PMID- 16715951 TI - Evaluation of the relationship between physiological FDG uptake in the heart and age, blood glucose level, fasting period, and hospitalization. AB - OBJECTIVE: Positron emission tomography (PET) with fluorodeoxyglucose (FDG) is widely used for evaluation of cancer and ischemic heart disease. Recently, increased myocardial FDG uptake has been reported to be related to some types of heart disease, such as sarcoidosis. However, the physiological increased FDG uptake in the heart often mimics the abnormal high uptake in these cases. In this study, we investigated the relationships between myocardial uptake and age, blood glucose level, fasting period, and hospitalization status (inpatient vs. outpatient). METHODS: A total of 159 non-diabetic patients were enrolled in the present study. Patients were imaged on a PET/CT scanner, and a three-dimensional region of interest (ROI) was drawn on the fused PET/CT image to measure the maximum standardized uptake value (SUV(max)) of the whole left ventricle. RESULTS: No significant relationships were observed between myocardial uptake and age or fasting period. Blood glucose level showed a significant relationship (p = 0.025) with myocardial uptake, but the R-square was extremely small (r2 = 0.03). With an SUV(max) threshold of 3.0, there was no significant difference between inpatients and outpatients. However, outpatients showed a significantly higher frequency of myocardial uptake over SUV(max) of 5.0 (chi2 test: p = 0.046). CONCLUSION: It is difficult to predict the degree of physiological uptake in the heart from data regarding age, fasting period, or blood glucose level. Outpatients tend to show higher myocardial uptake than inpatients, which may make it difficult to detect abnormally increased uptake in the heart. A long fasting period, such as overnight fasting, is an inadequate means to reduce the physiological uptake of FDG in the heart. PMID- 16715952 TI - Diastolic blood pressure influences cerebrovascular reactivity measured by means of 123I-iodoamphetamine brain single photon emission computed tomography in medically treated patients with occlusive carotid or middle cerebral artery disease. AB - OBJECTIVE: Impaired cerebrovascular reactivity (CVR) to vasodilating agents is a predictor of the onset and prognosis of ischemic stroke. It is realized that the CVR improves or worsens when measured periodically during the clinical course in medically treated patients with occlusive cerebrovascular disease. In these patients, we investigated the possible relationship between the interval change in CVR and that in systemic blood pressure (BP). METHODS: Forty-two patients (14 females and 28 males, mean age +/- SD: 65.3 +/- 8.8 years) with severe stenosis or occlusion of the common carotid, internal carotid, or middle cerebral arteries repeatedly underwent single photon emission computed tomography (SPECT) studies using 123I-iodoamphetamine to measure cerebral blood flow (CBF) distribution and CVR at a more-than-6-month interval (mean +/- SD: 18.5 +/- 8.8 months). The CVR was separately estimated in cerebral hemispheres ipsilateral and contralateral to the most severe vascular lesion as the % increase in CBF after acetazolamide loading to CBF at rest. Systemic BP was measured four times at enrollment and the follow-up SPECT studies during resting and acetazolamide loading. Average BP at each SPECT study was an average of BP measurements during resting and acetazolamide loading. Interval changes in CVR were correlated with those in average systolic BP, average diastolic BP, and average mean arterial BP. RESULTS: The interval changes in CVR were significantly correlated with those in average diastolic BP in the ipsilateral hemisphere (y = 0.71x + 1.43, r2 = 0.11, p < 0.05) and in the contralateral hemisphere (y = 0.88x - 0.46, r2 = 0.16, p < 0.05) but not with those in average systolic BP or average mean arterial BP. CONCLUSIONS: In medically treated patients with steno-occlusive carotid artery or middle cerebral artery lesions, the interval change in CVR to acetazolamide by means of 123I-IMP SPECT was influenced by the diastolic BP at the SPECT studies. Monitoring diastolic BP is important to evaluate interval change in CVR. PMID- 16715953 TI - Osteoid osteoma simulating osteomyelitis: differentiation with Tc-99m HIG scintigraphy. AB - We present a case of osteoid osteoma with a history of mild pain, local swelling and point tenderness on the right lower leg. The diagnosis of osteoid osteoma was difficult due to the atypical clinical history and misleading radiological and bone scan findings. When it is difficult to differentiate an osteoid osteoma from osteomyelitis using CT, MRI or bone scan; HIG scintigraphy can be used to exclude an infection. PMID- 16715954 TI - FDG PET scan in a primitive neuroectodermal tumor. AB - Primitive peripheral neuroectodermal tumor (PNET) is a rare tumor that often arises in soft tissue. Magnetic resonance imaging is a good diagnostic tool that can establish the extent of disease. The utility of positron emission tomography (PET) using F-18-fluoro-2-deoxy-D-glucose (FDG) in the diagnostic work-up and staging of PNET has not been well established. We present a case of PNET of the right upper extremity that did not show FDG uptake despite its large size and aggressive nature. PMID- 16715955 TI - SPECT in periodic lateralized epileptiform discharges (PLEDs): a case report on PLEDs. AB - Periodic lateralized epileptiform discharges (PLEDs), which are known as unusual electroencephalogram (EEG) patterns, are described in a patient who had stroke and seizures. This patient underwent Tc-99m HMPAO (hexamethyl propylene amine oxime) brain single photon emission computed tomography (SPECT) imaging both during PLEDs on EEG and after the cessation of PLEDs. The initial SPECT study revealed increased CBF in the left frontal and parietal cortex extending through the left temporal region and in the left basal ganglium. After the PLEDs disappeared, the second SPECT study showed decreased perfusion on the left frontal and parietal region in the brain. Brain SPECT findings supported the contention that PLEDs may be an ictal phenomenon. Here we also present a review on PLEDs and contributions of brain SPECT as a functional imaging modality to investigate the underlying mechanism of this interesting EEG pattern. PMID- 16715956 TI - Diaphyseal medullary stenosis with pleomorphic malignant fibrous histiocytoma of the bone: 99mTc hydroxymethylenediphosphonate and 201Tl chloride scintigraphy findings. AB - Diaphyseal medullary stenosis (DMS) is an extremely rare hereditary bone dysplasia, which was first described by Arnold in 1973. DMS has a high incidence of pleomorphic malignant fibrous histiocytoma (MFH). In this paper, we report the imaging findings of DMS with pleomorphic MFH of the bone, mainly describing 99mTc hydroxymethylenediphosphonate (HMDP) and thallium-201 (201Tl) chloride scintigraphy findings. On 99mTc HMDP scintigraphy, focal increased uptake area of the right femur corresponded to the area of bone marrow invasion of the tumor and bone infarction. The mechanism of the uptake of 99mTc HMDP to the extraosseous lesion was not clear. On 201Tl chloride scintigraphy, the increased uptake of the periphery of the mass seemed to reflect the aggressiveness of invasion and the cellularity. PMID- 16715957 TI - Effects of image reconstruction algorithm on neurotransmission PET studies in humans: comparison between filtered backprojection and ordered subsets expectation maximization. AB - OBJECTIVES: Both reconstruction algorithms, filtered backprojection (FBP) and ordered subsets expectation maximization (OSEM), are widely used in clinical positron emission tomography (PET) studies. Image reconstruction for most neurotransmission PET scan data is performed by FBP, while image reconstruction for whole-body [18F]FDG scan data is usually performed by OSEM. Although several investigators have compared FBP and OSEM in terms of the quantification of regional radioactivity and physiological parameters calculated from PET data, only a few studies have compared the two reconstruction algorithms in PET studies that estimate neurotransmission, i.e., neuroreceptor and neurotransporter binding. In this study we compared mean regional radioactivity concentration in the late phase and binding potential (BP) between FBP and OSEM algorithms in neurotransmission PET studies for [11C]raclopride and [11C]DASB. METHODS: Dynamic PET scans with [11C]raclopride in 3-dimensional mode were performed on seven healthy subjects. Dynamic PET scans with [11C]DASB in 2-dimensional mode were performed on another seven subjects. OSEM images were post-filtered so that its transverse spatial resolution became similar to that of FBP with the same Hanning filter (Kernel FWHM 6 mm). In both PET studies we calculated the BP of [11C]raclopride and [11C]DASB by a reference tissue model for each ROI (region of interest). RESULTS: There was no significant difference in mean regional radioactivity concentration between FBP and OSEM for [11C]raclopride and [11C]DASB. Only +2.4 - +3.2%, but still a significant difference in BP of [11C]raclopride between FBP and OSEM was observed in the striatum. There was no significant difference in BP between FBP and OSEM in other than the striatum for [11C]raclopride and in all regions for [11C]DASB. In addition, there was no significant difference in root mean square error between FBP and OSEM when BP was calculated. CONCLUSIONS: The BP values were similar between FBP and OSEM algorithms with [11C]raclopride and [11C]DASB. This study indicates that OSEM can be used for human neurotransmission PET studies for calculating BP although OSEM was not necessarily superior to FBP in the present study. PMID- 16715958 TI - System design and development of a pinhole SPECT system for quantitative functional imaging of small animals. AB - Recently, small animal imaging by pinhole SPECT has been widely investigated by several researchers. We developed a pinhole SPECT system specially designed for small animal imaging. The system consists of a rotation unit for a small animal and a SPECT camera attached with a pinhole collimator. In order to acquire complete data of the projections, the system has two orbits with angles of 90 degrees and 45 degrees with respect to the object. In this system, the position of the SPECT camera is kept fixed, and the animal is rotated in order to avoid misalignment of the center of rotation (COR). We implemented a three dimensional OSEM algorithm for the reconstruction of data acquired by the system from both the orbitals. A point source experiment revealed no significant COR misalignment using the proposed system. Experiments with a line phantom clearly indicated that our system succeeded in minimizing the misalignment of the COR. We performed a study with a rat and 99mTc-HMDP, an agent for bone scan, and demonstrated a dramatic improvement in the spatial resolution and uniformity achieved by our system in comparison with the conventional Feldkamp algorithm with one set of orbital data. PMID- 16715959 TI - [Hematopoietic and endothelial cell development from primate embryonic stem cells in vitro]. PMID- 16715960 TI - [Establishment of a novel molecular-targeted therapy for hematological malignancies mediated through reactive oxygen species (ROS)]. PMID- 16715961 TI - [The present status of, and problems with the development of FLT3 kinase inhibitors]. PMID- 16715962 TI - [State of the art in the treatment of chronic leukemias]. PMID- 16715964 TI - [New therapeutic strategy for Philadelphia chromosome-positive acute lymphoblastic leukemia]. PMID- 16715963 TI - [Recent advances in the clinical use of cytokines]. PMID- 16715965 TI - [Evaluation of the clinical relevance of serum measurements of free-light chains in patients with multiple myeloma]. AB - Measurement of serum free-light chains (FLCs) was performed using a recently developed immunoassay in 180 healthy individuals, 16 patients with multiple myeloma, and each 1 patient with Waldenstrom's macroglobulinemia, primary amyloidosis, or MGUS (monoclonal gammopathy of undetermined significance) to evaluate the clinical relevance of FLCs in the diagnosis and disease monitoring. Serum FLC levels of each patient were elevated compared with the levels of healthy individuals. The changes in FLC levels after treatment and at the time of relapse occurred earlier than those in serum immunoglobulin levels due to the short half-life of FLCs. Serum FLC levels were elevated in patients with Bence Jones type myeloma in spite of negative urinary immunoelectrophoresis. The serum FLC assay is a sensitive and useful tool for the diagnosis and monitoring of multiple myeloma and other B-cell proliferative disorders. PMID- 16715966 TI - [Ischemic colitis following the treatment of acute hemorrhage in a patient with acquired hemophilia A]. AB - A 66-year-old man with hypertension and hyperlipidemia developed a hemorrhagic stomal ulcer and massive hematoma of the face at 4 and 7 months, respectively, after fundusectomy for early gastric cancer. The diagnosis of acquired hemophilia A was made based on the marked prolongation of activated partial thromboplastin time, an extremely low factor VIII activity, and a very high-titer factor VIII inhibitor. After admission, oral prednisolone and cyclophosphamide were started. In addition, activated prothrombin complex concentrates and recombinant activated factor VII were intravenously administered which successfully controlled his hemorrhage. Only 1 week after the episode of bleeding, however, he complained of abdominal pain accompanied by watery stool with fresh blood. The diagnosis of ischemic colitis was made based on the clinical course and the findings on both CT-scan and colon fiberoscopy. The colitis spontaneously and quickly resolved with conservative observation. To the best of our knowledge, this is the first reported case of ischemic colitis that occurred in an acquired hemophilia patient without simultaneous administration of coagulation factors or antifibrinolytic agents. We should thus pay attention to the possible occurrence of thrombotic events even in acquired hemophilia patients in the presence of risk factors for thrombosis. PMID- 16715967 TI - [Role of CRAM (CRMP5) in growth cone development and semaphorin-mediated signaling]. PMID- 16715968 TI - [Hematopoietic stem cell niche in bone marrow]. PMID- 16715969 TI - [Structure and function of a PB1 domain, a novel cell polarity regulator]. PMID- 16715970 TI - [Regulation of cell cycle in pancreatic beta-cell and development of diabetes]. PMID- 16715972 TI - [Hepatitis C virus replication associated with lipid rafts]. PMID- 16715971 TI - [Biosynthetic enzymes responsible for sex pheromone production in Lepidoptera]. PMID- 16715973 TI - [Activity-dependent gene expression for neuronal plasticity]. PMID- 16715974 TI - [Role of NF-kappaB activation in inflammation-associated carcinogenesis]. PMID- 16715975 TI - [Alpha4 integrins control front-rear polarity during cell migration]. PMID- 16715976 TI - [Laparoscopic procedures in focal lesions of the liver]. AB - Seventy-three laparoscopic procedures were performed for the treatment of focal lesions of the liver. Fifty-four patients had operations for non-parasitic cysts of the liver and cystic disease. Polycystic disease of the liver was in 8 patients. Multiple cysts were in 19 patients, solitary - in 27. A total of 139 cysts were treated. Surgery was performed with the standard technique. Atypical marginal resection of the liver was performed in 16 patients for hemangioma (n=8), nodular hyperplasia (n=2), hepatic metastases (n=5), hamartoma (n=1). Atypical marginal resection by type of peritumor resection was conducted in 5 cases, and it was similar to segmentectomy in 11 cases. Laparoscopic cryodestruction of benign tumors and hepatic metastases was performed in 3 patients including in combination with atypical marginal resection in 2 cases. There were no intrasurgical complications which required laparotomies. There were no hemotransfusions. Long-term results were followed-up from 1 to 3 years in 25 patients with hepatic cysts and from 3 months to 3 years in 7 patients after marginal resection for hemangiomas and fibronodular hyperplasia. Ultrasonic examination revealed no recurrences of the disease. PMID- 16715977 TI - [Comparative assessment of short-term outcomes of cholecystectomy in minimally invasive surgical procedures for chronic calculous cholecystitis]. AB - A randomized blind trial was carried out for comparative evaluation of short-term results of surgical treatment of chronic calculous cholecystitis in 100 patients after laparoscopic cholecystectomy and in 100 patients after minimally-invasive cholecystectomy. Both groups contained geterogenous patients (morphology in gall bladder zone, concomitant diseases). Statistically significant (p=0.000001) decrease of hospital stay was revealed after laparoscopic cholecystectomy. It is necessary to keep exact selection criteria for each type of elective surgery in cholelithiasis. PMID- 16715979 TI - [Urgent laparoscopic surgeries in children of the first year of life]. AB - One hundred and ninety-two urgent laparoscopic operations in 179 infants (including 31 newborn infants) were performed. Indications for surgery were intestinal obstruction (108 children), acute diseases of abdominal organs (61), and gynecological diseases (10). The rate of conversion in 3 groups was 10.9, 4.6% and 0, respectively. In all the cases conversion was performed due to technical difficulties, there were no intrasurgical complications, diagnostic value of the method reached 100%. Combined laparoscopic surgeries were performed in 25.1% cases. Intestinal invagination was the most frequent cause of laparoscopy. Comparative analysis of open (28) and laparoscopic (85) desinvagination demonstrated the advantages of the latter one both during and after surgery. Laparoscopy is the useful method for diagnosis and treatment of acute abdominal disease requiring urgent surgery in infants. PMID- 16715978 TI - [Immune status late after surgery for spleen injuries]. AB - Immune status was studied in patients who had undergone surgery for spleen injuries (splenectomy, autolientransplantation and organ-saving surgeries). It is demonstrated that splenectomy leads to immunosuppression in long-term postoperative period. For prevention of these complications the organ-saving surgeries or autolientransplantation in addition to splenectomy are recommended. PMID- 16715980 TI - [Prediction of postoperative commissural complications in urgent abdominal surgery in children]. AB - Acetyl phenotype was examined in 166 patients with appedicular peritonitis and in 96 patients with acute adhesive intestinal obstruction. The age of the patients ranged from 5 months to 14 years. It was revealed that postoperative adhesive complications developed mostly in patients with fast acetylation (more than 76%). It is demonstrated that acetyl phenotype may be used as a screening-test for prediction of the risk of postsoperative adhesive complications and preventive treatment. PMID- 16715981 TI - [Transillumination phlebectomy in minimally-invasive surgical treatment of varicose disease]. AB - Transillumination phlebectomy with TriVex was used in 72 patients for the surgical treatment of lower extremities varicose veins. Diagnostic protocol, indications and surgical technique are described in detail. Short- and long-term results are assessed. Transillumination guarantees clear visualization of subcutaneous veins. It is an effective method in surgical treatment of varicose disease, permits to decrease number and length of incisions, time of surgery, to achieve good functional and esthetic results. PMID- 16715982 TI - [Results of percutaneous sclerotherapy of benign nodular affections of the thyroid gland with ethanol]. AB - The results of experimental (150 animals) and clinical (783 patients) studies of percutaneous sclerotherapy of benign nodular affections of the thyroid gland with ethanol are analyzed. Reduction of the cysts depends first of all on ratio of fluid to solid components. Technical features of the method and disputable points of mechanism of action are discussed. High efficacy of sclerotherapy in benign nodular affections of the thyroid gland is demonstrated. PMID- 16715983 TI - [Diagnostic and treatment program in acute adhesive obturative obstruction of the small bowel]. AB - Clinical classification of acute adhesive obturative obstruction of the small bowel is presented. Four stages of acute obstruction are distinguished. The proposed treatment and diagnostic program permitted to predict the course of obstruction, specify indications for surgical and conservative treatment, determine the terms of preoperative preparation. This program permitted to decrease the number of operated patients to 16% and decrease lethality in acute adhesive obturative obstruction of the small bowel to 1,67%. PMID- 16715984 TI - [Necrosis of epiploic appendages of the colon]. AB - Fifty-two patients with necrosis of epiploic appendages (EA) of the colon were treated. Necrosis of EA of the appendix was diagnosed in 4 (7,7%) patients, caecum - in 7 (13,5%), ascending colon - in 5 (9,6%), descending colon - in 2 (3,8%), and sigmoid colon - in 34 (65,4%) cases. Multiple necrosis was revealed in 3 (5,8%) patients. Surgical treatment of these patients is radical and justified. It prevents progress of destruction and development of complications. Long-term results are evaluated for the first time. PMID- 16715985 TI - [On the more secure intestinal sutures]. PMID- 16715986 TI - [Intraoperative ultrasound investigation and laser spectroscopy for diagnosis of cancer in patients with lung tumors]. AB - The experience in more than 2000 surgeries on the lungs is analyzed. Recently intraoperative ultrasonic and laser autofluorescent spectroscopic revision of tumors of the lungs and mediastinal organs is successfully used. Intrasurgical ultrasonic examination of the lungs permits to detect the structure and diffusion of primary lung tumor (invasion of vessels) that influences policy of treatment. Optical spectroscopy permits one to carry out intraoperative rapid diagnosis of benign and malignant lung tumors. PMID- 16715987 TI - [Instrument for formation of endosurgical nodes]. PMID- 16715988 TI - [Hemangiopericytoma of the lungs in a patient with angioleiomyolipoma of the kidney]. PMID- 16715989 TI - [Treatment of a patient with multiple intestinal fistulas complicated with migration of obturation device]. PMID- 16715990 TI - [Prognostic factors in surgical treatment of diffuse toxic goiter]. PMID- 16715991 TI - [The problem of postoperative pyoseptic complications in abdominal trauma with spleen injury in terms of immunological disorders]. PMID- 16715992 TI - [Mechanical systems of assisted circulation]. PMID- 16715993 TI - [Academician E. N. Meshalkin (the 90th anniversary of birth)]. PMID- 16715994 TI - Promotion of occupational health and safety research: foundation of a new independent administrative institution in Japan. PMID- 16715995 TI - Prevalence of respiratory symptoms among workers in industries of south Tehran, Iran. AB - OBJECTIVE: The aim of the present study was to determine the prevalence of respiratory symptoms from occupational lung hazards among workers in industries of south Tehran, IRAN. METHODOLOGY: This was a cross-sectional study in which by multistage random sampling items on demographic characteristics, cigarette smoking, occupational history and respiratory symptoms were collected of workers. RESULTS: The mean age of the workers was 38.5 (SD = 10.2) yr: age ranged from 19 to 70 yr. Of 880 workers under study, 252 (28.7%) were smoking. Also, it has been observed that workers exposed in the workplace with occupational chemical exposures such as dust, gas and fume pollutants. The prevalence of respiratory symptoms was cough (20.7%), phlegm (41.6%), dyspnea (41.7%), feel tightness (27.4%) and nose irritation (23.5%). CONCLUSIONS: Occupational exposures among workers in industries of south Tehran may cause respiratory symptoms and respiratory disorders, engineering controls and industrial hygiene is recommended. PMID- 16715996 TI - Indoor glutaraldehyde levels in the endoscope disinfecting room and subjective symptoms among workers. AB - We measured the environmental glutaraldehyde (GA) levels during the disinfection of endoscopes and also investigated the subjective symptoms of the workers engaged in that work. At 6 hospitals in the Tokyo and Kanagawa area, 8 rooms for endoscope washing and disinfecting the endoscopy equipment were surveyed. The geometric mean environmental GA levels in the 8 rooms were 1.3 to 19.6 ppb. The personal exposure levels at the time of replacing the antiseptic solution containing GA in two of the disinfecting rooms were 94.2 and 84.9 ppb. Subjective symptoms such as ophthalmic, nasal, respiratory, pharyngeal symptoms and nausea were more prevalent among workers than controls as evidenced from the questionnaire survey. PMID- 16715997 TI - Carcinogenicity and chronic toxicity of 1,4-dichloro-2-nitrobenzene in rats and mice by two years feeding. AB - Carcinogenicity and chronic toxicity of 1,4-dichloro-2-nitrobenzene (DCNB) were examined by feeding each group of 50 F344 rats and 50 BDF1 mice of both sexes a DCNB-containing diet at a concentration of 0 (control), 320, 800 or 2,000 ppm (w/w) for 2 yr. In rats, incidences of hepatocellular adenomas and carcinomas and their combined incidence were increased in the 2,000 ppm-fed males, together with increased incidence of basophilic cell foci in the 800 and 2,000 ppm-fed males. A dose-related increase in combined incidences of renal cell adenomas and carcinomas was noted. Incidence of Zymbal gland adenomas tended to increase in the 2,000 ppm-fed males. In mice, incidences of hepatocellular adenomas in the 800 and 2,000 ppm-fed females and hepatocellular carcinomas in the 2,000 ppm-fed males and in the 800 and 2,000 ppm-fed females were increased. Incidence of hepatoblastomas was increased in all DCNB-fed males and in the 2,000 ppm-fed females. Signs of chronic toxicity were characterized by centrilobular hypertrophy of hepatocytes with nuclear atypia in mice, increased relative liver weight in rats, a dose-related increase in incidences of chronic progressive nephropathy with advanced grades of severity in male rats, and decreased hemoglobin concentration and hematocrit accompanied by increased bone marrow hematopoiesis in female rats. Carcinogenic activity of DCNB was evaluated for the three different tumors, and sensitive signs of the chronic toxicity were dis- PMID- 16715998 TI - Overtime work, cigarette consumption, and addiction to cigarette among workers subject to mild smoking restrictions. AB - The goal of the present study was to investigate the relation of hours of overtime work to cigarette consumption and addiction to cigarette, which was measured by the heaviness of smoking index. The subjects were 571 male daily smokers who responded to a cross-sectional survey of municipal employees of a Japanese city office, in which smoking was permitted in designated areas. Those who engaged in moderate overtime work (10-29 h per month) consumed less number of cigarettes per day and had lower levels of heaviness of smoking index, compared with those who worked either shorter or longer hours of overtime, although the differences were not statistically significant. In the workplace, men who worked 50 h or longer overtime last month consumed, on average, 4 cigarettes more than men who worked less than 30 h of overtime. Home cigarette consumption decreased as hours of overtime work increased. In stratified analysis, there was a significant difference in daily cigarette consumption according to hours of overtime work among smokers in staff position or under low psychological work stress; showing reduced consumption associated with medium levels of overtime work, compared to either no overtime work or extended overtime hours. The U shaped relations of hours of overtime work to overall cigarette consumption and addiction to smoking deserve further investigations. PMID- 16715999 TI - Physical work load affects the maximum oxygen uptake. AB - PURPOSE: Maximum oxygen uptake (VO2max) is known to be influenced by life-style factors, such as exercise and smoking. We aimed to further test the hypothesis that, besides these, work form also contributes to increase VO2max among workers. METHOD: Three thousand six hundred and forty eight male workers with age 38.3 +/- 12.1 and 1,575 female workers with age 35.6 +/- 11.1, ranging from 20-year old to 69-year old, who participated in Total Health Promotion Plan at workplaces in Fukui Prefecture in 1998 were selected. Data on VO2max were analyzed for age, systolic blood pressure (SBP), body mass index (BMI), work form (sedentary, standing or ambulatory), exercise, and smoking. RESULTS: Multiple regression analysis showed that work form ("standing", "ambulatory"), and exercise habits might increase VO2max while BMI, age, and SBP might decrease VO2max in male participants. Exercise habits were suggested to increase VO2max while BMI, SBP, and age might decrease VO2max in females. In females smoking, was suggested to increase VO2max. CONCLUSION: After adjusting for age, BMI, SBP, exercise, and smoking, it was suggested that the physical work load, represented by the work form, may contribute to increases in VO2max in males. Implications of smoking among females with respect to VO2max is discussed. PMID- 16716000 TI - Associations between lifestyle and mental health in a group of Japanese overseas workers and their spouses resident in Dusseldorf, Germany. AB - This study investigated associations between lifestyle factors and selected aspects of mental health in a group of Japanese overseas workers and their accompanying spouses who were residing in and around Dusseldorf, Germany, in February 1994. Considering four aspects of mental health (depression, mental instability, nervousness and neurosis) and six lifestyle factors (alcohol consumption, sleeping hours, cigarette smoking, physical exercise, eating breakfast and eating snacks), a cross-sectional study involving 822 volunteers (486 workers and 336 spouses) was performed using the Todai Health Index (THI) for surveying self-perceived health and a lifestyle related self-administered questionnaire. Alcohol consumption had no associations with any of the four aspects of mental health, and only very weak inverse associations were found between the other five lifestyle factors and the four aspects of mental health in the workers group. In the spouses group, physical exercise was the only lifestyle factor significantly associated with mental health. PMID- 16716001 TI - Relation of average and highest solvent vapor concentrations in workplaces in small to medium enterprises and large enterprises. AB - The present study was initiated to examine the relationship between the workplace concentrations and the estimated highest concentrations in solvent workplaces (SWPs), with special references to enterprise size and types of solvent work. Results of survey conducted in 1010 SWPs in 156 enterprises were taken as a database. Workplace air was sampled at > or = 5 crosses in each SWP following a grid sampling strategy. An additional air was grab-sampled at the site where the worker's exposure was estimated to be highest (estimated highest concentration or EHC). The samples were analyzed for 47 solvents designated by regulation, and solvent concentrations in each sample were summed up by use of additiveness formula. From the workplace concentrations at > or = 5 points, geometric mean and geometric standard deviations were calculated as the representative workplace concentration (RWC) and the indicator of variation in workplace concentration (VWC). Comparison between RWC and EHC in the total of 1010 SWPs showed that EHC was 1.2 (in large enterprises with>300 employees) to 1.7 times [in small to medium (SM) enterprises with < or = 300 employees] greater than RWC. When SWPs were classified into SM enterprises and large enterprises, both RWC and EHC were significantly higher in SM enterprises than in large enterprises. Further comparison by types of solvent work showed that the difference was more marked in printing, surface coating and degreasing/cleaning/wiping SWPs, whereas it was less remarkable in painting SWPs and essentially nil in testing/research laboratories. In conclusion, the present observation as discussed in reference to previous publications suggests that RWC, EHC and the ratio of EHC/WRC varies substantially among different types of solvent work as well as enterprise size, and are typically higher in printing SWPs in SM enterprises. PMID- 16716002 TI - Involvement of thyroxine in ovarian toxicity of di-(2-ethylhexyl) phthalate. AB - Forced ovulation induced by the administration of exogenous gonadotropin is a useful marker for studying the ovarian toxicity of chemicals in experimental animals. We examined the toxicity of di-(2-ethylhexyl) phthalate (DEHP) in the ovaries of immature F344 female rats. Superovulation was induced by injections of equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG) in rats dosed with 125, 250, 500, 1,000 or 2,000 mg/kg body weight of DEHP for 4 consecutive days. The number of ova shed during superovulation significantly decreased in rats treated with DEHP at 500 mg/kg as compared with control, but no changes were observed in the number of ova in groups given other doses of DEHP. In control rats treated with olive oil, hypophysectomy reduced significantly the number of ovulated ova. When 2,000 mg DEHP was given to hypophysectomized (hypox) rats, the number of ova in the hypox group was significantly smaller than that in the intact group administered with the same doses of DEHP. In contrast, the numbers of ova of the intact and hypox groups did not significantly differ in rats given 500 mg DEHP. The levels of circulating thyroxine (T4) were significantly decreased by 2,000 mg DEHP in intact rats, and a tendency for T4 to decrease in T4 was also observed in hypox rats given 2,000 mg DEHP. These results suggest that daily administration of 500 mg DEHP suppressed superovulation in immature F344 rats by disrupting the hypothalamic-pituitary-ovarian axis in a manner similar to that of hypophysectomy. Decreased circulating T4 levels seemed to negate this disruption as observed in recovered superovulation after treatment with 2,000 mg DEHP. PMID- 16716003 TI - Tentative criteria for assessing workers exposure to toluene by urinary toluene screening. AB - This study assessed screening thresholds for determining workers exposure to toluene (Tol) by urinary Tol (Tol-U) and proposed applicable criteria for on-site settings. Participants' urine samples (n = 21) were collected at the end of the workday during the latter half of a week and the Tol-U concentration was assayed. Simultaneously, each worker's exposure dose to Tol in the breathing zone during work, Tol-TWA (time-weighted average), was measured. Tentative criteria were proposed. Level I, less than Tol-U 38 microg/l, has the least chance of exceeding Tol-OEL 50 ppm (occupational exposure limit for Tol recommended by the Japan Society for Occupational Health), probability 95% <. Level II, Tol-U 38-60 microg/l, has a low possibility of exceeding Tol-OEL. Level III, Tol-U 60-110 microg/l, has a high possibility of exceeding Tol-OEL. Level IV, more than Tol-U 110 microg/l, clearly exceeds Tol-OEL, probability 95% <. PMID- 16716004 TI - Field study to explore possible effects of styrene on auditory function in exposed workers. AB - OBJECTIVES: We conducted this study to examine, whether occupational styrene exposures are associated with reduced hearing ability. METHODS: The auditory function was investigated by pure tone audiometry and registration of transitory evoked otoacoustic emissions (TEOAE) in 32 workers of a fibre-reinforced plastic boat building factory. Sixteen subjects were laminators (mean age: 41 yr (SD: 8)) and therefore regularly exposed to styrene with mean duration of exposure to styrene of 7.5 yr (SD 5.0). The tests were applied to a reference group of 16 workers (mean age: 39 yr (SD: 8)) who were not directly exposed to styrene but had a similar noise exposure. RESULTS: A few and isolated correlations between the parameters of hearing acuity and exposure indices, such as current internal styrene exposures (sum of MA and PGA) and duration of styrene exposure, were statistically significant, but no consistent association was found. CONCLUSION: The results of this study do not support the assumption of an ototoxic effect of chronic styrene exposure in workers. PMID- 16716005 TI - Field evaluation of mercury vapor analytical methods: comparison of the "double amalgam method" and ISO 17733. AB - In this study, a gold amalgam method called the "Double amalgam method" was compared with the ISO 17733 method for mercury vapor analysis method. In terms of sensitivity and ease of operation, the amalgamation method is superior to the oxidation method. Two parallel samplings were carried out in this research at a button battery factory, where the mercury vapor level in the air was about 0.001 mg/m3 and at a fluorescent lamp factory, where the mercury vapor level was about 0.015 mg/m3. In the both cases, the measured values of the two showed good agreement with each other. As these two workplaces represent typical mercury levels in industries today, the double amalgam method is applicable to working environment measurement. PMID- 16716006 TI - Assessment of subjective stress in video display terminal workers. AB - Stress assessment in the workplace has been focused on its environmental, psychological and biological aspects. We carried out an evaluation of the subjective components of stress in a working population of 60 subjects employed in a large Public Service, 30 Video Display Terminal (VDT) workers (15 men and 15 woman) and 30 office-workers not assigned to VDT (15 men and 15 woman), by using the "Rapid Stress Assessment Scale": a short questionnaire of easy administration in work environment. VDT workers of both sexes showed higher total stress score vs. office workers (respectively p<0.05, p<0.05). Gender differences were present: female VDT workers showed higher scores of clusters anxiety (p<0.001) and aggressiveness (p<0.05); male VDT workers' score were significantly higher in somatization (p<0.05) and aggressiveness cluster. Our results showed that in VDT workers are experienced greater subjective response to stress than "office workers" and confirm the gender differences in stress experiencing. PMID- 16716007 TI - Significant improvement from chronic beryllium disease following corticosteroid pulse therapy. AB - Chronic beryllium disease (CBD) is a rare disease characterized by diffuse interstitial pulmonary granulomatosis. We report a case of CBD which exhibited marked improvement both subjectively and objectively following pulse therapy. The patient was a 36-year-old man whose chief complaint was dyspnea and a dry cough. Since July 1990, the patient had been working in the development of an automatic or mechanical technique for producing beryllium-copper alloy. It appeared likely that the patient may have been exposed to metal beryllium fumes generated from an opening located just above the furnace. The Be concentration exceeded 25 microg/m3 transiently in the breathing zone in this workplace. A chest X-ray film taken in October 1994 showed fine granular shadows throughout the entire lung fields. Around August 1998, the patient's dyspnea became aggravated. An X-ray taken at that time showed linear and reticular shadows, in addition to the diffuse fine granular shadow. In October 1998, after 3 days of methylprednisolone pulse therapy, oral prednisolone 30 mg was initiated. With this treatment, the patient's pulmonary function tests and blood gases improved. Once the patient's condition had improved sufficiently, the dosage of prednisolone was decreased by 2.5 mg every two weeks. The patient continues to be monitored. PMID- 16716008 TI - Air conditioning systems as non-infectious health hazards inducing acute respiratory symptoms. AB - Chronic and acute exposure to toxic aerosols belongs to frequent causes of airway diseases. However, asthma attacks due to long-distance inhalative exposure to organic solvents, transmitted via an air condition system, have not been reported so far. The present case illustrates the possibility of air conditioning systems as non-infectious health hazards in occupational medicine. So far, only infectious diseases such as legionella pneumophila pneumonia have commonly been associated to air-conditioning exposures but physicians should be alert to the potential of transmission of toxic volatile substances via air conditioning systems. In view of the events of the 11th of September 2001 with a growing danger of large building terrorism which may even use air conditioning systems to transmit toxins, facility management security staff should be alerted to possible non-infectious toxic health hazards arising from air-conditioning systems. PMID- 16716009 TI - Relationship of blood lead levels to blood pressure in exhaust battery storage workers. AB - Several researches has focused the hypothesis that low blood lead levels could be associated with an increased risk of hypertension. To assess the relation between occupational lead exposure and elevated blood pressure a group of 27 workers, age range from 27 to 62 years, mean (SD) 36.52 (+/- 8.16) yr; length of employment mean (DS) 2.97 (+/- 1.67) yr, were recruited as study subjects. The following variables were measured: blood lead concentration (BPb), delta-Aminolevulinic Acid Dehydratase (ALAD) activity, Zinc Protoporphirin (ZPP), creatinine, hematocrit, Body Mass Index (BMI) and Systolic Blood Pressure (SBP) and Diastolic Blood (DBP) Pressure. The results showed that long term occupational exposure was related to a slight increase of systolic and diastolic blood pressure among workers who had been exposed to higher level of lead with respect to workers exposed to lower level of lead. Furthermore, blood lead concentration (BPb) and ZPP resulted higher among workers exposed to higher level of ambient lead, while in the same group of workers ALAD activity resulted more inhibited. The authors concluded long term cumulative lead exposure can significantly increase blood pressure in low level Pb exposed workers. PMID- 16716010 TI - Exposure to organic solvents among handicraft car painters: A pilot study in Italy. AB - Car repair painters usually experience long-term exposure to many different solvents. In Italy, the greater part of car painting shops are "handicraft", i.e. 2-5 workers and small premises. Usually workers do not have specific duties, but everyone takes part in all different operations. Moreover, working time is not standardized but varies according to the workload, working methods tend to be traditional, and compliance to individual protection devices is poor. We have hence assessed, on a sample of 8 italian handicraft car painting shops, the exposure levels to solvents, implementing three classic exposure monitoring methods: environmental sampling with charcoal tubes, personal sampling with diffusive charcoal samplers, and urinary determination of unmetabolised solvents. A simple regression analysis was performed to evaluate relationships between the three series of data. The solvents analysed were toluene, ethylbenzene, 1, 2 dichloropropane, n-butylacetate, n-amylacetate, xylene isomers, ethylacetate, and benzene. Benzene was included due to several factors, among others its presence (1% v/v) in the italian unleaded gasoline. Benzene was found in all shops, at levels around or higher than the 8-h time-weighted average limit (8-h TLV-TWA). Other solvents were found in the different shops at various levels, 10(-2) - 10( 1) times the 8-h TLV-TWA. Air concentrations of toluene, n-butylacetate, xylenes, and benzene were positively correlated with urinary levels of the parent compounds, while a negative correlation was found for ethylbenzene. The health implications of workers' exposure level was briefly discussed. PMID- 16716011 TI - Cross-sectional analysis of blood lead level of entire Korean lead workers. AB - We analyzed blood lead (PbB) level from the 2003 health surveillance results of 13,043 lead workers from 1,217 total lead industries to evaluate lead intoxication at low level in Korea. Geometric mean of PbB was 6.08 microg/dl and 56.6% and 7.9% of total lead workers had PbB level over than 5 microg/dl and 25 microg/dl, respectively. Male showed relatively higher PbB level compared to women, but "Manufacture of Other Electronic Valves, Tubes and Electronic Components n.e.c." had more women than male in risk of low level lead exposure. While conventional high-risk industry such as "Manufacture of Accumulators" and "Other Basic Non-ferrous Metal Industries" were remained in high-risk group in lead exposure, there were high risks in other industries such as plastic, chemical and part manufacturing. Non-production tasks such as fork lift truck driving, maintenance, lab testing, and supporting function showed high blood lead level in addition to routine manufacturing processes such as smelting and soldering. PMID- 16716012 TI - Influence of tilt training on activation of the autonomic nervous system in patients with vasovagal syncope. AB - OBJECTIVE: Tilt training is a new treatment for vasovagal syncope. Its therapeutic efficacy is thought to be the result of the desensitization of cardiopulmonary receptors, but it could be the influence of the tilt training on the activation of the autonomic nervous system as well. METHODS AND RESULTS: The study group consisted of 24 vasovagal patients (17 women and 7 men) aged 32.5 +/- 11.8 years. The diagnostic head-up tilt test was performed according to the Italian protocol with nitroglycerin if necessary. The monitoring head-up tilt test was performed according to the Westminster protocol without provocation, after 1 to 3 months of tilt training. Holter ECG recordings for HRV parameters (time and frequency domain) were obtained from selected 2-min intervals before, during and after the diagnostic and monitoring tilt test. The diagnostic test was positive in the passive phase in 6 and after provocation in 18 patients. During the training period no syncope occurred. Analysing the HRV parameters we demonstrated the following findings: I. mRR decreases immediately after assumption of a vertical position in both tests (diagnostic and monitoring) but in the diagnostic test its further decrease occurs earlier than in the monitoring test; 2. the absolute power of the HF component is greater in the early phase of tilt after tilt training than in the corresponding period in the diagnostic test. CONCLUSIONS: After a longer period of tilt training the activation of the sympathetic nervous system in response to the erect position is diminished. PMID- 16716013 TI - The usefulness of tissue doppler imaging in the determination of left ventricular diastolic filling pressure. Comparison with other techniques. AB - The further examination of left ventricular diastolic function (LVDF) is important in terms of early diagnosis, therapy and follow-up of heart failure. Although cardiac catheterization is considered as the most accurate method in the evaluation of diastolic function, since it is invasive, noninvasive methods are preferred. This study was undertaken to examine whether mitral annular velocities assessed by pulsed tissue Doppler imaging (PTDI) were associated with invasive measures of diastolic LV pressure and whether additional information was gained over traditional transmitral and pulmonary venous flow velocity parameters. Doppler examination was performed in 102 patients referred to our clinic for cardiac catheterization. Doppler signals from the mitral inflow, pulmonary venous inflow, and PTDI of the mitral annulus were obtained. Mean left ventricular diastolic pressure (M-LVDP) was measured. The relationship between echocardiographic parameters and M-LVDP was investigated.A significant correlation was observed between M-LVDP and E/septal Em (r = 0.52, p < 0.000), E/lateral Em (r = 0.45, p < 0.0001), E/posterior Em (r = 0.46, p < 0.0001) E/anterior Em (r = 0.49, p < 0.0001), E/mean Em (r = 0.49, p < 0.0001), PVadur - MVadur (r = 0.51, p < 0.0001). The best echocardiographic parameters correlating with M-LVDP were E/septal Em and PVa-dur - MVa-dur. In conclusion, PTDI could be used in the assessment of LVDP. However, the combination of transmitral flow and pulmonary venous flow velocities with annular velocity can be proposed as the best method for assessing LV filling pressure that combines the influence of transmitral driving pressure and myocardial relaxation. PMID- 16716014 TI - Diabetes and impaired fasting glucose as predictors of morbidity and mortality in male coronary artery disease patients with reduced left ventricular function. AB - OBJECTIVES: To evaluate the prognostic value of impaired fasting glucose and diabetes mellitus in male patients with coronary artery disease and poor left ventricular function. METHODS AND RESULTS: From a prospective database on patients referred for gated myocardial perfusion imaging between 1998 and 2002 all male patients with a history of coronary artery disease and poor left ventricular function were selected. Poor function was defined as left ventricular ejection fraction < or = 40%. Subjects were classified as non-diabetics with fasting blood glucose levels < 110 mg/dL, non-diabetics with impaired fasting glucose (fasting blood glucose between 110 and 125 mg/dL) and diabetics. Median follow-up was 2.7years. End points were all-cause mortality, cardiac death and hospitalization for heart failure. One hundred and sixty patients were selected (age 65 +/- 9 years and left ventricular ejection fraction 29 +/- 8%). In univariate analysis atrial fibrillation, NYHA class, glycaemia and diabetes mellitus discriminated between survivors and non-survivors. In Cox multivariate regression analysis for all-cause mortality only NYHA class and diabetes mellitus remained significant. Kaplan Meier analysis showed that diabetics had the worst survival and non-diabetics with glucose < 110 mg/dL had the best survival. Non diabetics with impaired fasting glucose had intermediate survival. Analysis for cardiac death/hospitalization for heart failure showed similar results. CONCLUSION: In male patients with coronary artery disease and impaired left ventricular function diabetes mellitus and fasting glucose are strongly predictive of poor outcome. Diabetics have the worst prognosis but non-diabetics with impaired fasting glucose also are at higher risk compared to nondiabetics with low fasting blood glucose. PMID- 16716015 TI - Augmentation of revascularization and prevention of plasma leakage by angiopoietin-1 and vascular endothelial growth factor co-transfection in rats with experimental limb ischaemia. AB - BACKGROUND: Angiopoietin-I (Ang I) and vascular endothelial growth factor (VEGF) are endothelial cell-specific growth factors. In the current study, we tested the hypothesis that co-expression of AngI and VEGF could have an effect on development of leakage-resistant vessels. METHODS AND RESULTS: Expression plasmids, pcD2 (control plasmid), pcD2/AngI, pcD2NEGF121, or pcD2/AngI +pcD2NEGF121, were injected intramuscularly into an ischaemic hindlimb rat model, followed by electroporation. Collateral vessel development and skeletal muscle atrophy were assessed before and 7, 14, 28 days after treatment. Capillary density was significantly increased in the rats transfected with AngI or VEGF compared with that in the rats transfected with pcD2 alone (P < 0.05). Rats transfected with AngI + VEGF had the highest capillary density (P < 0.05). The mean perimeter ratio of ligated hindlimb to non-ligated hindlimb was lower with pcD2 treatment rats compared with that in the rats transfected with AngI, VEGF or AngI +VEGF (P < 0.01). Limb necrosis was observed in two of 33 control rats, but none in the AngI and/orVEGF gene-transferred rats. The amount of extravasated Evans blue in rat ligated hindlimb muscle (7 days after treatment) treated with both AngI and VEGF was significantly less compared with that in the rats transfected with VEGF alone (P < 0.01). CONCLUSIONS: The current study demonstrated that co-expression of AngI and VEGF genes in the ischaemic muscle effectively develops leakage-resistant vessels in the rat model. Therefore, this approach may provide a more appropriate therapeutic strategy in ischaemic vascular diseases. PMID- 16716016 TI - Inflection point of ascending aortic waveform is a predictive factor for major adverse cardiac events after successful coronary stent placement in acute myocardial infarction. AB - OBJECTIVE: The reflection waveform in the ascending aortic pressure has been reported to reflect systemic arterial stiffness, and increase the risk of coronary heart disease. The purpose of this study is to evaluate prospectively whether the reflection waveform in the ascending aortic pressure is associated with the risk of major adverse cardiac events (MACE) after successful coronary stenting in acute myocardial infarction (AMI). METHODS AND RESULTS: One hundred and twenty-five patients with AMI and undergoing successful coronary stenting were enrolled in this study. We measured the inflection time to determine the reflection waveform in the ascending aortic pressure at angioplasty. Inflection time was defined as the time interval from initiation of a systolic pressure waveform to the inflection point. During a period of 2945 person-months, we confirmed 42 cases of MACE. Shorter inflection time increased incident MACE. Kaplan-Meier analysis demonstrated a significantly reduced event-free rate in patients with inflection time < or = 86 ms (MACE; p < 0.01, cardiovascular death; p < 0.001 by log-rank test). The multiple-adjusted hazard ratio for a I standard deviation (SD) magnitude decrease in inflection time was 1.46 (95% confidence interval, 1.03 to 2.05) for MACE, and 5.76 (95% confidence interval, 1.78 to 18.67) for cardiovascular death. CONCLUSIONS: Shorter inflection time increased the risk of MACE after successful coronary stenting in patients with AMI. PMID- 16716017 TI - Pro-inflammatory cytokines and their receptors in chronic heart failure: do they really matter? AB - The pathophysiological insights into the chronic heart failure (CHF) syndrome and, consequently, the management of patients with CHF have dramatically changed over the past few decades. Antagonists of the activated neurohormonal pathways are now a mainstay in the therapeutic arsenal. It seems, however, that a neurohormonal ceiling might have been reached. Other humoral mal-adaptations have recently gained importance. One of them is the presence of a low-grade inflammatory status characterising patients with moderate to severe CHF. The presence, both at tissue level and in the circulation, of increased levels of pro inflammatory cytokines suggests that immune activation might be a relevant mechanism contributing to cardiac as well as peripheral manifestations of the disease. Whereas targeted anti-cytokine treatment has proven unsuccessful in the CHF population, the concept of immune modulation is still intensively studied. PMID- 16716018 TI - Dilated cardiomyopathy in childhood: prognostic features and outcome. AB - The natural history of dilated cardiomyopathy in children is difficult to predict due to the heterogeneous character of the disease. The outcome in infants and children is highly variable from complete recovery to death. In this study, 40 children diagnosed with dilated cardiomyopathy between 1995 and 2004 in our paediatric cardiology unit were reviewed with respect to clinical course and outcome, retrospectively. The medical history of these patients with dilated cardiomyopathy was reviewed to determine age, gender, family history, preceding viral illness, duration of symptoms before the diagnosis, symptoms and signs at presentation, treatment and outcome. The diagnosis was made on the basis of cardiomegaly and evidence of poor left ventricular function by echocardiography. Median age at diagnosis was 14 months, ranging from 2 months to 8 years. At presentation, 28 patients (70%) were under and twelve (30%) were above the age of two years. Twenty-eight (70%) patients had signs of congestive heart failure. Mean duration of follow-up was 40 +/- 24 months (ranging from 6 months to 9 years), 21 patients (52.5%) recovered, 17 patients (42.5%) had residual disease and two (5%) died. The cause of death in both patients was progressive cardiac failure. Sixteen of 28 patients (57%) who were below the age of two years and five of 12 patients (42%) who were above the age of two years at presentation recovered. The rate of recovery was significantly different between the two age groups (p < 0.05). Seventeen of 21 (81%) patients with a history of recent viral illness at presentation recovered. The mean duration of the disease among those who recovered was 11 +/- 8.3 months. Five of 19 (26%) patients without recent viral illness recovered. The mean duration of the disease in this group was 22 +/ 12 months. There was a significant difference between the two groups with respect to recovery and recovery time (p < 0.05). During the first 6 months after diagnosis, there was a significant difference between the patients who recovered and the patients who had residual disease with respect to improvement in the left ventricular FS (22 +/- 3.5%, 15.2 +/- 2.8%, respectively) (p < 0.05). In conclusion, in this study, the rate of recovery and survival is higher than in previous studies. A good outcome is related to age at presentation (< or = two years old), a history of viral disease within three months of presentation and improvement in ventricular function during the first 6 months after diagnosis. Intractable heart failure has an adverse effect on the outcome. PMID- 16716019 TI - Incidence and complication rate of infective endocarditis in the Dutch region of Walcheren: a 3-year retrospective study. AB - OBJECTIVE: To measure the incidence of infective endocarditis (IE) in the region of Walcheren and to estimate the complication and comorbidity rate of IE, we conducted a 3-year retrospective study of IE in the only hospital (Ziekenhuis Walcheren) of the region between January, 2002 and December, 2004. RESULTS: The total number of cases was 32. The calculated yearly incidence of IE was 9.61 cases per 100.000 inhabitants per year. The median age was 64 years (range 36-81 years). When applying the Duke criteria 28 patients (87.5%) were classified as definite IE and 4 patients (12.5%) as possible IE. Blood cultures were positive in 27 patients (84.4%). The most commonly isolated organisms were streptococci (37.5%). Staphylococcus aureus was isolated in 31.3% of positive blood cultures. Enteroccocus faecalis was identified in 3 cases (9.4%). In only 15.6% of the cases was the course of IE uncomplicated. The most frequent complications were heart failure (59.4%) and embolic events (34.4%). Cardiac surgery was performed in 37.5% of the cases. Concomitant morbidity was found in 75% of the patients. Especially, the high incidences of diabetes mellitus (28.1%), chronic renal failure (28.1%) and chronic obstructive pulmonary disease (21.9%) were remarkable. CONCLUSION: The calculated incidence of IE of 9.61 cases per 100,000 inhabitants per year was more than five times higher than the one reported in a nation-wide Dutch study of 1992. The present study demonstrates that IE remains a disease with a considerable mortality and complication rate. The majority of the patients with IE had non-cardiac comorbidity. Especially, the prevalence of diabetes mellitus and chronic renal failure in our population of patients with IE is remarkable. PMID- 16716020 TI - Tilt training: a new challenge in the treatment of neurally mediated syncope. AB - Clinical experience with repeated tilt testing and standing training has shown that this procedure can be used as a new therapy for the restoration of abnormal autonomic orthostatic reflexes. The introduction of tilt training therapy for neurally mediated syncope is a new and effective treatment. This new therapy has been used recently in several centres with excellent results. Since syncope is a severe social handicap for the patient, successful therapy will normalise the functional status and restore self-confidence. The use of tilt training can be considered as first-line therapy in this disorder. In the majority of the studies on tilt training, patients remain free of syncope during active treatment with this therapy. In about 50% of the patients with neurally mediated syncope already the second consecutive tilt test became negative. This therapeutic effect of in hospital repeated tilt tests is sustained by continued standing training at home. PMID- 16716021 TI - Salmonella paratyphi endocarditis. AB - Endocarditis is a rare complication of salmonellosis. The most frequently observed species is Salmonella typhi. Endocarditis caused by Salmonella paratyphi is extremely rare with only three previous cases documented in the medical literature. We report a case of endocarditis caused by Salmonella paratyphi in an otherwise healthy young patient, with full recovery after appropriate antibiotic treatment. PMID- 16716022 TI - Atrial fibrillation in hypertrophic cardiomyopathy: another indication for circumferential pulmonary vein ablation? AB - Atrial fibrillation is often highly symptomatic in patients with hypertrophic cardiomyopathy. In the present paper we describe the results of circumferential pulmonary vein ablation in two such patients. Although multiple antiarrhythmic drugs failed to prevent recurrences of AF in each patient, catheter ablation resulted in long-term maintenance of sinus rhythm and stabilization of heart failure. PMID- 16716023 TI - Living with a congenital aortic stenosis. PMID- 16716024 TI - Congenital valvular aortic stenosis. surgical treatment. PMID- 16716025 TI - Genetics of left ventricular outflow tract abnormalities. PMID- 16716026 TI - Percutaneous interventions for congenital aortic stenosis. PMID- 16716027 TI - A new approach to heart valve tissue engineering. PMID- 16716028 TI - Left heart in the fetus: planning of pregnancy. PMID- 16716029 TI - The role of imaging techniques in diagnosis and evaluation of congenital aortic stenosis. PMID- 16716030 TI - Congenital left heart outflow abnormalities in the newborn. PMID- 16716031 TI - Heart valve tissue engineering. PMID- 16716032 TI - Medication use among pregnant women. AB - OBJECTIVES: This study examines medication use among women aged 15 to 49, comparing pregnant women with their nonpregnant contemporaries. A portrait of women who used medication during pregnancy is also presented. DATA SOURCES: Analysis is based on data from the 1994/95 through 2002/03 National Longitudinal Survey of Children and Youth (NLSCY), as well as the 2003 Canadian Community Health Survey (CCHS) and the 1996/97 National Population Health Survey (NPHS). ANALYTICAL TECHNIQUES: Estimates of medication use are based on cross-sectional data. Logistic regression was used to determine factors associated with use of medication. MAIN RESULTS: Medication use among women aged 15 to 49 has risen over the last 10 years. Although the proportion of pregnant women who used medications increased, it remained below the figure for other women the same ages. Over one quarter of women (27%) were taking prescription medications while pregnant. PMID- 16716033 TI - Survival from cancer--up-to-date predictions using period analysis. AB - OBJECTIVES: This period analysis provides Canadian predictions of the short- and long-term relative survival of people recently diagnosed with cancer. Long-term period and cohort-based estimates are also compared. DATA SOURCES: Data are from the Canadian Cancer Registry, the Canadian Mortality Data Base, and Statistics Canada life tables. ANALYTICAL TECHNIQUES: Relative survival analyses were conducted using the life-table method; expected survival proportions were derived using the Ederer II approach. Period analysis estimates were based on the survival experience of cancer cases followed up in 2002. The cohort analyses involved people diagnosed in 1997 (5-year survival) or 1992 (10-year survival). National estimates exclude Quebec. MAIN RESULTS: Relative survival ratios were highest for thyroid (5-year, 97.7%) and prostate (95.2%) cancer and lowest for pancreatic cancer. Survival for many forms of cancer is higher than previously estimated by cohort-based analysis. The largest increases in 10-year relative survival were predicted for cancers of the prostate (13.0%) and rectum (9.7%). The largest predicted increases for 5-year survival were for cancers of the cervix uteri (5.4%) and rectum (4.5%), and for leukemia (3.7%). PMID- 16716034 TI - The effect of universal influenza immunization on vaccination rates in Ontario. AB - OBJECTIVES: This article examines the association between introduction of Ontario's Universal Influenza Immunization Program and changes in vaccination rates over time in Ontario, compared with the other provinces combined. DATA SOURCES: The data are from the 1996/97 National Population Health Survey and the 2000/01 and 2003 Canadian Community Health Survey, both conducted by Statistics Canada. ANALYTICAL TECHNIQUES: Cross-tabulations were used to estimate vaccination rates for the total population aged 12 or older, for groups especially vulnerable to the effects of influenza, and by selected socio demographic variables. Z tests and multiple logistic regression were used to examine differences between estimates. MAIN RESULTS: Between 1996/97 and 2000/01, the increase in the overall vaccination rate in Ontario was 10 percentage points greater than the increase in the other provinces combined. Increases in Ontario were particularly pronounced among people who were: younger than 65, more educated, and had a higher household income. Between 2000/01 and 2003, vaccination rates were stable in Ontario, while rates continued to rise in the other provinces. Even so, Ontario's 2003 rates exceeded those in the other provinces. PMID- 16716035 TI - Flu shots--national and provincial/territorial trends. PMID- 16716036 TI - Formalin produces depolarizations in human airway smooth muscle in vitro. AB - Respiratory irritants may result in airway smooth muscle (ASM) depolarization and bronchoconstriction. We examined the effect of formalin on membrane potentials in human ASM in two types of in vitro preparations: strip preparations, which contain functional sensory and motor nerve endings and cultured cells, which lack these nerve endings due to the tissue dissociation process. Depolarizations occurred in atropine-treated strip preparations in response to formalin exposures, but not in similarly-treated cultured cells, suggesting a role for non cholinergic mediators in formalin-induced depolarization. It is suggested that formalin may act as an irritant to produce bronchoconstriction that is mediated by the release of endogenous substance P (SP) from peripheral sensory nerve endings. This is supported by our observation that exogenous SP produced depolarizations of a magnitude similar to those produced by formalin in both strip preparations and cultured cells. In addition, capsaicin, which releases endogenous SP from nerve endings, produced depolarizations of a magnitude similar to formalin in strip preparations, but was without effect in cultured cells. PMID- 16716037 TI - Health effects of subchronic exposure to diesel-water-methanol emulsion emission. AB - The U.S. Environmental Protection Agency's National Ambient Air Quality Standards for ozone and particulate matter (PM) require urban non-attainment areas to implement pollution-reduction strategies for anthropogenic source emissions. The type of fuel shown to decrease combustion emissions components versus traditional diesel fuel, is the diesel emulsion. The Lubrizol Corporation, in conjunction with Lovelace Respiratory Research Institute and several subcontracting laboratories, recently conducted a health assessment of the combustion emissions of PuriNOx diesel fuel emulsion (diesel-water-methanol) in rodents. Combustion emissions from either of two, 2002 model Cummins 5.9L ISB engines, were diluted with charcoal-filtered air to exposure concentrations of 125, 250 and 500 microg total PM/m3. The engines were operated on a continuous, repeating, heavy-duty certification cycle (U.S. Code of Federal Regulations, Title 40, Chapter I) using Rotella-T 15W-40 engine oil. Nitrogen oxide (NO) and PM were reduced when engines were operated on PuriNOx versus California Air Resources Board diesel fuel under these conditions. Male and female F344 rats were housed in Hazleton H2000 exposure chambers and exposed to exhaust atmospheres 6 h/day, five days/week for the first 11 weeks and seven days/week thereafter. Exposures ranged from 61 to 73 days depending on the treatment group. Indicators of general toxicity (body weight, organ weight, clinical pathology and histopathology), neurotoxicity (glial fibrillary acidic protein assay), genotoxicity (Ames assay, micronucleus, sister chromatid exchange), and reproduction and development were measured. Overall, effects observed were mild. Emulsion combustion emissions were not associated with neurotoxicity, reproductive/developmental toxicity, or in vivo genotoxicity. Small decreases in serum cholesterol in the 500-microg/m3 exposure group were observed. PM accumulation within alveolar macrophages was evident in all exposure groups. The latter findings are consistent with normal physiological responses to particle inhalation. Other statistically significant effects were present in some measured parameters of other exposed groups, but were not clearly attributed to emissions exposure. Positive mutagenic responses in several strains of Salmonella typhimurium were observed subsequent to treatment with emulsion emissions subfractions. Based on the cholesterol results, it can be concluded that the 250-microg/m3 exposure level was the no observed effect level. In general, biological findings in exposed rats and bacteria were consistent with exposure to petroleum diesel exhaust in the F344 rat and Ames assays. PMID- 16716039 TI - Levels of retinyl palmitate and retinol in stratum corneum, epidermis and dermis of SKH-1 mice. AB - Vitamin A (retinol) regulates many biological functions, including epidermal cell growth. Retinyl palmitate (RP) is the major esterified form of retinol and the predominant component of retinoids in the skin; however, how endogenous levels of RP and retinol in the skin are affected by the age of the animal remains unknown. Furthermore, the levels of retinol and RP in the various skin layers - the stratum corneum, epidermis and dermis of skin - have not been reported. In this paper, we report the development of a convenient method for separation of the skin from SKH-1 female mice into the stratum corneum, epidermis, and dermis and the determination of the levels of RP and retinol in the three fractions by HPLC analysis. The total quantities of RP and retinol from the stratum corneum, epidermis, and dermis are comparable to those extracted from the same amount of intact skin from the same mouse. There was an age-related effect on the levels of RP and retinol in the skin and liver of female mice. An age-related effect was also observed in the stratum corneum, epidermis, and dermis. The levels of RP and retinol were highest in the epidermis of 20-week-old mice, and decreased when the age increased to 60- and 68-weeks. The total amount of RP at 20 weeks of age was found to be 1.52 ng/mg skin, and decreased about 4-fold at 60- and 68-weeks of age. A similar trend was found for the effects of age on the levels of retinol. PMID- 16716038 TI - Concentrations of Ni and V, other heavy metals, arsenic, elemental and organic carbon in atmospheric fine particles (PM2.5) from Puerto Rico. AB - Fine atmospheric particulate PM2.5 (particles with diameters of < 2.5 microm) were sampled in an urban industrialized area--Guaynabo, Puerto Rico (Figure 1)- and in a reference less polluted site Fajardo, Puerto Rico--and analyzed for trace metals, and inorganic and organic elemental carbon. PM2.5 samples were collected from November 2000 to September 2001 using an Andersen Instruments RAAS2.5-400 for periods of 72 h. Metals analyzed were arsenic (As), cadmium (Cd), copper (Cu), iron (Fe), nickel (Ni), lead (Pb), vanadium (V) and zinc (Zn) by atomic absorption. Levels of elemental and organic carbon (EC/OC) were also determined. All metals analyzed, except for Fe, were significantly higher in PM2.5 from Guaynabo when compared to Fajardo. Average levels of PM2.5 in Guaynabo were 11.6 versus 8.5 microg/m3 in Fajardo. Average levels of EC were 1.5 and <0.14 microg/m3; and OC levels were 2.2 and < 1 microg/m3 for Guaynabo and Fajardo, respectively. Levels of Ni (17 ng/m3) and V (40 ng/m3) determined in PM2.5 from the Guaynabo area were high when compared to other cities, and these metals could be responsible for respiratory problems reported in the area. Multivariate analyses showed strong relationships in Guaynabo between Ni and V, PM2.5 and Fe and As and Cu and Pb. In Fajardo, the strongest associations were obtained between PM2.5 and Fe, Cd and V and Ni and Pb and Cu, these last three elements exhibiting an inverse relationship. PMID- 16716040 TI - Thrombocytosis induced in mice after subacute and subchronic V2O5 inhalation. AB - Reports about vanadium (V) inhalation toxicity on the hematopoietic system, specifically about coagulation are limited. Therefore, we decided to evaluate the effects of V with a complete blood count and morphologic analysis of platelets on blood smears. CD-1 male mice inhaled V2O5 0.02 M 1 h twice weekly over 12 weeks. Blood samples were obtained by direct heart puncture; Wright stained smears were used for platelet quantification. An increase in platelet count from the third week of exposure was observed, as well as the presence of megaplatelets. Our results demonstrate, for the first time, that V induces thrombocytosis and it might correlate with some thromboembolic diseases. Further analysis is needed to evaluate the functionality of these platelets as well as the cause of its increase. PMID- 16716041 TI - Antioxidant enzyme activities and lipid peroxidation products in heart tissue of subacute and subchronic formaldehyde-exposed rats: a preliminary study. AB - OBJECTIVE: The aim of this experimental study was to evaluate the oxidant/antioxidant status and lipid peroxidation in the heart of rats exposed to formaldehyde (FA) inhalation for four weeks (subacute) or 13 weeks (subchronic) continuously. METHODS AND RESULTS: Sixty Wistar albino rats were divided into six groups randomly (ten in each group). The first and second groups were used as subacute and subchronic control groups. FA gas was generated from paraformaldehyde and pumped to a closed glass chamber. Rats were exposed to atmosphere containing 10 and 20 ppm FA (8 h/day, five days per week) during a four and 13 weeks period. After heart tissues were obtained and homogenized, thiobarbituric acid-reactant substances (TBARS) and nitric oxide (NO) levels, as well as superoxide dismutase (SOD) and catalase (CAT) activities, were measured. There were statistically significant findings in SOD and CAT activities in the study groups compared to the control group. Heart tissue SOD level was increased in the group exposed to subacute 10 and 20 ppm FA inhalation compared to the control group (P < 0.011 and <0.0001). In addition, heart tissue SOD level was increased in the group exposed to subchronic 10 and 20 ppm FA inhalation compared to the corresponding control group (P < 0.001). On the other hand, there were statistically significant decreases in CAT activity in subacute 10 and 20 ppm groups compared to the corresponding control group (P < 0.012 and < 0.039, respectively). Although not significant, TBARS levels were increased in both subacute 10 ppm (P = 0.100) and subchronic 20 ppm (P = 0.053) groups compared to their corresponding control groups. Tissue NO levels were unchanged upon FA inhalation. In the correlation analyses, a meaningful relationship between SOD and CAT activities in subchronic 10 ppm group (r = -0.685, P < 0.029); SOD activity and TBARS level in subchronic 20 ppm group (r = -0.675, P < 0.032); and CAT activity and NO level in subchronic 20 ppm group (r = -0.810, P < 0.005) were found. CONCLUSION: From the findings of our study, it can be interpreted that subacute and subchronic FA inhalation may stimulate oxidative stress and thus, some secondary toxic effects in cardiac cells and tissue. This increase in the oxidative stress could not induce lipid peroxidation in the membranous structure of cardiac cells. An increased SOD enzyme activity was thought to be secondary to decreased CAT activity, as a compensation mechanism, preventing heart tissue from destruction induced by FA. PMID- 16716042 TI - The effects of ginkgo biloba extract on tissue adenosine deaminase, xanthine oxidase, myeloperoxidase, malondialdehyde, and nitric oxide in cisplatin-induced nephrotoxicity. AB - This study was carried out to determine if Ginkgo Biloba Extract (GBE or Egb 761) exerts a beneficial effect against cisplatin-induced renal failure in rats. Sprague Dawley rats were divided into four groups. The first group (control) received orally 1 mL/kg/day of 0.9% saline by an oral carrier vehicle on days 1 to 10. The second group was injected with 7 mg/kg cisplatin intraperitoneally (i.p.) on the fourth day, once only. The third group (vit E+cisplatin) was administered 10 mg/kg/day i.p. vit E on 1 to 10 days with one dose of i.p. cisplatin (7 mg/kg) injection on the fourth day. The fourth group (GBE+cisplatin) was given GBE orally at 100 mg/mL/kg started on the first day up to the tenth day with one dose of cisplatin (7 mg/kg) injection on the fourth day. Cisplatin was found to lead a statistically significant increase in plasma BUN and creatinine levels, as well as urine micro total protein (MTP) levels, leading to acute renal failure (ARF) in rats. Renal xanthine oxidase (XO) activities increased in all groups (statistically significant in cisplatin + GBE-treated rats; P < 0.001). Adenosine deaminase (AD) activities were increased in cisplatin-treated rats, and decreased in cisplatin+GBE-treated (P < 0.041) and cisplatin+vit E-treated (P < 0.005) rats, compared to controls. Malondialdehyde (MDA), nitric oxide (NO) levels and myeloperoxidase (MPO) activities were increased in the kidney tissue of cisplatin-treated rats. Vit E improved plasma creatinine and urine MTP levels, together with tissue MDA, NO levels, and MPO activities. But GBE had no statistically significant effect on those parameters. These results indicate that increased XO, AD and MPO activities, as well as MDA and NO levels play a critical role in cisplatin nephrotoxicity. GBE has been shown to protect against cisplatin induced nephrotoxicity. PMID- 16716043 TI - Effects of dioxins on the quantitative levels of immune components in infants. AB - Dioxins (polychlorinated dibenzo-p-dioxin (PCDD)+polychlorinated dibenzofuran (PCDF)) and polychlorinated biphenyls (PCBs) are potentially hazardous compounds and have structural similarity with thyroid hormones. Animal studies have demonstrated that PCDDs, PCDFs and PCBs can alter immune functions. However, in humans it is not yet elucidated whether dioxins contained in breast milk have any effects on the immune functions in infants. To investigate the effects of dioxins on the immune system, we compared the quantitative levels of immune components between a breast-fed group and bottle-fed group, in which dioxin concentration is almost zero. Ratios of immune cells, such as CD4+ and CD8+ T-lymphocytes, as well as B-lymphocytes (CD19+ and/or CD20+) and NK cells (CD16+, CD56+) in peripheral blood lymphocytes, serum immunoglobulin level, and level of specific IgE antibody to allergens in the venous blood at 12 months of age were assessed in a subgroup of 281 infants. The relationship of post-natal dioxin exposure via breast feeding with the ratio of immunological markers and the level of humoral antibodies up to 12 month of age was not demonstrated. In conclusion, it would appear that the content of dioxins in breast milk in the Japanese general population is not enough to induce any change in theses-examined immunological parameters during the first year of life, although long-term effects remain to be evaluated. PMID- 16716044 TI - Influence of thyroxine and n-propylthiouracil on nephro-toxicity of inorganic arsenic in rat. AB - The effect of hyper or hypoactive thyroid on the renal toxicity of arsenic trioxide has been studied in rats. It was observed that pre-treatment of rats with thyroxine stimulates arsenic excretion in urine. The anti-thyroid drug n propylthiouracil (PTU), inhibits the accumulation of arsenic in renal tissue. Both treatments affect the renal pathology. Histopathological lesions are less severe in PTU and arsenic-treated rats in comparison to thyroxine and arsenic treated rats. Ultrastructural studies support light microscopical observations. An adaptive response was noticed against arsenic in PTU pre-treated rats. We attribute this response to decreased glutathione-S-transferase (GSH) activity and increased GSH synthesis in the kidney. A relationship between thyroidal activity and arsenic toxicity is suggested by present observations. PMID- 16716045 TI - [How the bovine viral diarrhea virus outwits the immune system]. AB - The interaction of bovine viral diarrhea virus (BVD virus) with its host has several unique features, most notably the capacity to infect its host either transiently or persistently. The transient infection stimulates an antiviral immune reaction similar to that seen in other transient viral infections. In contrast, being associated with immunotolerance specific for the infecting BVD viral strain, the persistent infection differs fundamentally from other persistent infections like those caused by lentiviruses. Whereas the latter are characterized by complex viral evasion of the host's adaptive immune response by mechanisms such as antigenic drift and interference with presentation of T cell epitopes, BVD virus avoids the immune response altogether by inducing both humoral and cellular immune tolerance. This is made possible by invasion of the fetus at an early stage of development. In addition to adaptive immunity, BVD virus also manipulates key elements of the host's innate immune response. The non cytopathic biotype of BVD virus, which is capable of persistently infecting its host, fails to induce type I interferon. In addition, persistently infected cells are resistant to the induction of apoptosis by double-stranded RNA and do not produce interferon when treated with this pathogen-associated molecular pattern (PAMP) that signals viral infection. Moreover, when treated with interferon, cells persistently infected with non-cytopathic BVD virus do not clear the virus. Surprisingly, however, despite this lack of effect on persistent infection, interferon readily induces an antiviral state in these cells, as shown by the protection against infection by unrelated viruses. Overall, BVD virus manipulates the host's interferon defense in a manner that optimises its chances of maintaining the persistent infection as well as decreasing the risks that heterologous viral infections may carry for the host. Thus, since not all potential host cells are infected in animals persistently infected with BVD virus, heterologous viruses replicating in cells uninfected with BVD virus will still trigger production of interferon. Interferon produced by such cells will curtail the replication of heterologous viruses only, be that in cells already infected with BVD virus, or in cells in which the heterologous virus may replicate alone. From an evolutionary viewpoint, this strategy clearly enhances the chances of transmission of BVD virus to new hosts, as it attenuates the negative effects that a global immunosuppression would have on the survival of persistently infected animals. PMID- 16716046 TI - Recent achievements and trends in the molecular diagnosis of bovine viral diseases--a view from the "OIE Collaborating Centre for the Application of Polymerase Chain Reaction Methods for the Diagnosis of Viral Diseases in Veterinary Medicine". AB - The activities of the "OIE Collaborating Centre for the Application of Polymerase Chain Reaction Methods for the Diagnosis of Viral Diseases" and its partner laboratories are summarised, providing a view of recent achievements and trends in the field the molecular diagnosis of bovine viral diseases. Results are briefly discussed concerning the diagnostic application of various techniques and approaches, such as PCR, semi-nested and nested PCR assays, phylogeny and molecular epidemiology, real-time PCR assays, "general" PCR systems, multiplex PCR, multi PCR, robotics in nucleic acid extraction, automated diagnosis, international standardisation and validation of the PCR-based diagnostic assays, and several future trends in molecular diagnostics. These achievements and trends lead to more rapid and specific detection of various viral infections in the cattle populations. Their application will certainly contribute to the reduction of losses which are caused by bovine viral diseases worldwide. PMID- 16716047 TI - [Genetic typing of classical swine fever viruses--a review]. AB - Classical swine fever (CSF) is a notifiable disease of domestic pigs and wild boar. It is caused by the highly contagious CSF virus and in its acute form the disease generally results in high morbidity and mortality. Due to the great economical impact an outbreak can cause to the pig industry it is one of the most important swine diseases worldwide. To limit the damage in the case of a new outbreak it is necessary to identify the virus as fast as possible. This information helps epidemiologists to trace the origin of the virus and to follow the virus spread. Genetic typing revealed that CSF virus genotypes, subgroups and types show a regional distribution making it an important tool for epidemiologists. Meanwhile, besides epidemiological data and nucleotide sequences from European isolates, information from isolates from South- and Central America, the Caribbean, Asia and recently from South Africa have become available. The data are stored in a database in the EU Reference Laboratory for CSF, accessible by the WWW (http://viro08.tiho-hanno ver.de). A new module was implemented that allows efficient automated genotyping. PMID- 16716048 TI - Replication of classical swine fever virus strains and isolates in different porcine cell lines. AB - Classical swine fever virus (CSFV) is an economically important pathogen of domestic pigs and wild boar. Due to the highly variable clinical picture of CSF, laboratory methods are essential for an unambiguous diagnosis. Virus isolation using cell culture is still considered the gold standard. It is based on the incubation of permissive cells with organ or leukocyte preparations followed by antigen detection. In the "EU Diagnostic Manual for CSF Diagnosis", the permanent cell line PK(15) (porcine kidney) is recommended. In the European Reference Laboratory (EURL) a clone of this cell line, PK(15)A, and the STE (swine testicular epitheloid) cell line are in use for propagation of CSFV. The aim of this work was to assess the relative ability of eleven permanent cell lines derived from various organs of wild boar and domestic pig, respectively, to support the replication of different strains and isolates in comparison to these cell lines. An avirulent and a highly virulent laboratory CSFV strain, and several recent field isolates from domestic pigs and wild boars were used. Titers were determined after one, two and three virus passages, and after 48 and 120 h of incubation. Of the eleven cell lines analyzed, two were found that replicated all the tested CSFV strains and field isolates. Those may be useful for improving diagnosis of CSFV and for preparing low-passaged virus stocks of new isolates. PMID- 16716049 TI - A versatile flow cytometry-based immunofluorescence inhibition assay for the detection of bovine viral diarrhea virus-specific antibodies. AB - A fast, sensitive and reliable flow cytometry-based (FACS = fluorescence activated cell sorting) immunofluorescence inhibition assay (FACS-IFI) for the detection of virus-specific antibodies in sera is described. The method was evaluated using sera from cattle experimentally infected with bovine viral diarrhea virus (BVDV). Virus-infected cells, which were fixed and permeabilized, were incubated with diluted sera from immunized or control animals. Monoclonal antibodies (mabs) against different viral proteins were added, and detected with ALEXA488-conjugated goat-antimouse antibodies. The fluorescence signals were detected by flow cytometry and determined as mean channel values. Results were expressed as percent fluorescence inhibition compared to standardized negative sera. The FACS-IFI test with sera from experimentally infected animals was highly sensitive and specific. Comparison of the FACS-IFI results with a commercially available blocking ELISA, an indirect ELISA and the standard serum neutralization test showed a strong correlation. Furthermore, the detection of protein-specific antibodies was possible using the FACS-IFI test. PMID- 16716050 TI - [Tick-borne encephalitis (TBE) with special emphasis on infection in horses]. AB - The tick-borne encephalitis (TBE), also known as early summer meningo encephalitis, is a geographically limited virus infection transmitted mainly by ticks. The importance of TBE is largely underestimated. The causative agent TBE Virus (TBEV) is grouped into the genus Flavivirus of the virus family Flaviviridae. Clinical disease including fatal outcomes has been described for men and dogs. With regard to horses only a limited number of case reports is available. In a study performed at the Institute of Virology, Justus-Liebig University Giessen serum samples from the German endemic region of Marburg Biedenkopf were tested for antibodies against TBEV. From 240 sera tested 7 (2.9%) were regarded as positive in a serum neutralization test (SNT). In an ELISA, performed in parallel to confirm the SNT results, 5 out of 7 positive sera from the SNT were also positive. The remaining two samples with low SNT-titres and all sera from horses negative in the SNT were also negative in the ELISA. This article is focussed on TBE of horses. In this context different aspects of TBE are included such as properties of the causative agent, interactions between causative agent, host animals and environment, spread of TBEV, pathogenesis, clinical symptoms, diagnosis and control. PMID- 16716051 TI - Roles of an extracellular matrix (ECM) receptor and ECM processing enzymes in demyelinating canine distemper encephalitis. AB - Canine distemper virus (CDV) belongs to the genus Morbillivirus of the Paramyxoviridae family. Due to the central nervous system (CNS) tropism of the virus and associated neuropathological changes, demyelinating canine distemper encephalitis (CDE) represents a relevant model for human demyelinating diseases like multiple sclerosis. The present review decribes the role of CD44 antigen (CD44), the principle cell surface receptor for hyaluronate and extracellular matrix (ECM) processing enzymes (matrix metalloproteinases [MMPs]) and their inhibitors (TIMPs) in the pathogenesis of demyelination. In acute and subacute CDE, a plaque-associated CD44 up-regulation is found that parallels astrocyte activation. Likewise, MMPs and TIMPs are prominently up-regulated in these lesions and are expressed mostly by astrocytes and microglia. In chronic lesions, CD44 expression declines together with the number of glial fibrillary acidic protein (GFAP) positive astrocytes. In addition, in this plaque type, CD44 is expressed on the cell membrane of perivascular mononuclear cells. In this phase, a decrease of MMP and TIMP expressions apart from MMP-11, -12, and -13 is obvious. In summary, CD44 and MMPs might be associated with the onset of demyelination and may interact to initiate ECM disturbances. Ligation of CD44 in the early phase may induce chemokines and cytokines and hence initiate and perpetuate the inflammatory process. In the chronic phase, it is conceivable that a MMP-TIMP imbalance may be the motor for lesion progression with a simultaneous influx of CD44-positive activated immune cells. PMID- 16716052 TI - Transmissible gastroenteritis virus infection: a vanishing specter. AB - About twenty years ago, a new coronavirus, porcine respiratory coronavirus (PRCoV), was detected in swine herds. This virus is related to transmissible gastroenteritis virus (TGEV); however, it is not enteropathogenic but causes only minor respiratory symptoms. As PRCoV shares some epitopes for neutralizing antibodies with TGEV, it acts like a nature-made vaccine against TGEV resulting in a drastic reduction of TGE outbreaks in Europe. A major difference between the two porcine coronaviruses is a large deletion in the surface protein S gene of PRCoV. Because of this structural difference, TGEV but not PRCoV has a sialic acid binding activity that allows the attachment to mucins and mucin-type glycoproteins. The sialic acid binding activity may allow TGEV to overcome the mucus barrier in the gut and to get access to the intestinal epithelium for initiation of infection. PMID- 16716053 TI - Case report: the significance of genotyping for the epidemiological tracing of classical swine fever (CSF). AB - In Germany, eleven outbreaks of CSF in domestic pig holdings were reported in 2002. They occurred exclusively in regions where CSF virus circulated in the wild boar population. In ten cases the phylogenetic analysis revealed that the isolates from domestic pigs and wild boar had identical sequences in the 5' non translated region (5'NTR). However, in one case a subtype was isolated which was slightly different from the virus subtype found in the wild boar population of that region. This case is decribed in detail. The epidemiological significance of different diagnostic methods is discussed, in particular the genetic typing of CSF virus isolates. PMID- 16716054 TI - [Case report: equine arteritis virus (EAV) as the cause of abortion in alpacas?]. AB - Here we report a case of a late abortion of a primiparous alpaca where genome fragments of the equine viral arteritis virus (EAV) could be detected in fetal tissues using reverse transcription polymerase chain reaction (RT-PCR). All five alpacas of the herd had virus neutralizing antibodies against EAV. EAV thus must be regarded as a potential agent for abortions in alpacas. Possible routes of introduction of the virus are discussed. PMID- 16716055 TI - Decompression illness presenting as breast pain. AB - We present two cases of decompression illness in women in whom the initial symptom causing distress after completion of the dives was breast pain. Both women were also subsequently found to have a patent foramen ovale. We postulate that breast pain may be an unusual under-recognized manifestation of decompression illness. PMID- 16716056 TI - Platelet function in humans is not altered by hyperbaric oxygen therapy. AB - A pilot survey of platelet function was performed on 6 patients undergoing hyperbaric oxygen therapy (2.0 ATA O2 for 2 hours, 6 days/week) for prophylaxis against osteoradionecrosis. Blood was drawn immediately prior to and after the first, tenth and twentieth treatment for measurements of platelet aggregation, ATP release and expression of activated alphalIb3 integrin. No significant differences were observed due to hyperbaric oxygen exposures. PMID- 16716057 TI - Monoplace hyperbaric chamber use of U.S. Navy Table 6: a 20-year experience. AB - We report a 20-year experience at LDS Hospital, Salt Lake City, UT using the U.S. Navy Treatment Table 6 (TT6) in an oxygen-filled monoplace hyperbaric chamber (1985-2004). Air breathing was provided via a demand regulator fitted with a SCUBA mouthpiece while the patient wore a nose clip. Intubated patients were mechanically ventilated with a Sechrist 500A ventilator, with a modified circuit providing air, when specified. We treated 90 patients: 72 divers (decompression sickness [DCS] = 67, arterial gas embolism [AGE] = 5), 10 hospital-associated AGE, and 8 miscellaneous conditions. They received a total of 118 TT6 (9 TT6 in intubated patients). Ninety-four percent of the TT6 schedules were tolerated and completed. The intolerance rate from two surveyed multiplace chambers was zero and 3% of 100 TT6 schedules each. Failure to complete the TT6 was due to oxygen toxicity (4) and claustrophobia (3). The U.S. Navy TT6 was well tolerated by patients with DCS or AGE treated in monoplace hyperbaric chambers, but tolerance may not be as high as when treated in the multiplace chamber. PMID- 16716058 TI - Proposing short-term observation units for the management of decompression illness. AB - Decompression illness (DCI) is a potentially life-threatening disease, often requiring hyperbaric oxygen therapy (HBO2) for symptom resolution. Once treated, current guidelines recommend an observation period of at least six hours for patients with neurological symptoms in case of relapse. Surveys have shown a symptom relapse rate as high as 38.5%, with half of those occurring in the first twenty-four hours. We propose that a short-term observation unit (OU) would be an ideal setting for these patients to be monitored. To evaluate this, we did a retrospective study of patients presenting with DCI at a major hyperbaric facility. One hundred and two consecutive patients were evaluated with DCI diagnosis and receiving HBO2. Forty-two (41.2%) patients had neurological sequelae; ten required more than one treatment for refractory symptoms or relapse. Thirty-eight of the forty-two patients received up to three treatments, which can be done within the time requirements of short-term observation. We conclude that OUs would provide a safe and efficient disposition for patients after receiving HBO2. PMID- 16716059 TI - Hyperbaric nitrogen and pentobarbital on synaptosomal membrane lipids and free fatty acids. AB - Nitrogen at high pressures and anesthetics increase lipid monolayer surface pressure and in turn modulates monolayer associated lipolytic enzyme activity that could alter membrane lipids. We tested the hypothesis that nitrogen at pressures of 5 and 10 megapascals (MPa) and pentobarbital induce alterations in synaptosomal membrane phospholipid and free fatty acid (FFA). Rat cortical synaptosomes in Krebs-Henseleit buffer were placed in steel chambers and incubated for four hours at 37 degrees C: at 5 or 10 MPa of O2/balance N2; at one 0.1 MPa on room air, and with 10 mg pentobarbital. Free fatty acids (FFA) were quantified by thin-layer and gas chromatography, and neutral and acidic lipids by high-pressure thin layer chromatography and protein by Biorad colorimetric assay. Statistical analyses were by ANOVA and posthoc analysis by Neuman-Keuls and Kruskal-Wallis tests at p < 0.05. Sphyngomyelin, phosphatidylcholine, phosphatidylethanolamine, cerebroside and cholesterol were unchanged by 5 and 10 MPa nitrogen and pentobarbital. Free fatty acids (16:00, 18:00, 18:01, 20:00, 22:0, 22:01 and 24:01) at 10 MPa were reduced compared to 5 MPa (p < 0.05) but unaffected by pentobarbital. The decrease in synaptosomal membrane FFA at 10 MPa suggests attenuated hydrolysis of membrane phospholipids without detectable alterations in membrane phospholipid composition. PMID- 16716060 TI - Post-exercise reduction in diffusing capacity of the lung after moderate intensity running and swimming. AB - A reduction in transfer factor of the lung for carbon monoxide (Tl(co)) is a consistent finding after saturation dives and is reported after some open sea air dives. Several diving associated factors may contribute to this reduction in Tl(co) including hyperoxia, venous gas microembolism, increased breathing resistance and immersion. Exercise, which inevitably is associated with open sea diving, may itself cause a reduction in Tl(co) up to at least 12 hours post exercise. Six trained swimmers and six trained runners who had never dived performed 30 min moderate intensity swimming and running on different days and in random order at approximately 75% of their maximal heart rate. Lung function including a flow-volume loop and Tl(co) was measured 30 min before and 60-90 min after exercise. There were no significant changes in dynamic lung volumes or maximal expiratory flow rates, but there was a reduction in Tl(co) of 4.5 (SD = 4.8) and 4.7 (SD = 4.6) % after swimming and running respectively (p < 0.01). There was no difference in this response between runners and swimmers, and the response was not associated with lung size. Even moderate exercise preceding measurements of Tl(co) should generally be accounted for, and this effect may contribute significantly to lung function changes immediately after open sea dives. PMID- 16716061 TI - Diffusing capacity and spirometry following a 60-minute dive to 4.5 meters. AB - The purpose of this study was to assess the contribution of SCUBA to the pulmonary effects of diving to 4.5 meters depth in healthy subjects using a randomized crossover control condition. Ten healthy divers performed two 60 minute 'dives' using SCUBA in a swimming pool. The non-immersed 1 ATA SCUBA control exposure took place at ambient pressure in the laboratory. Thirty minutes prior to, and 30 and 90 minutes post-exposure, FVC (forced vital capacity), FEV1.0 (forced expired volume), peak expiratory flow rate (PEFR), diffusing capacity (DL(co)), heart rate (HR) and temperature were measured. No significant differences were noted in HR, temperature or spirometry between the two conditions. A significant reduction in diffusing capacity occurred at 30 and 90 minutes after the pool dive (9.3% and 15.1%, respectively, p < 0.05). There was no concordant change in DL(co) following the non-immersed 1 ATA SCUBA control. Thus, a pool dive to 4.5 meters for 60 minutes causes a decrease in DL(co), without a change in spirometry, while breathing from SCUBA equipment without immersion causes no significant change in lung function. PMID- 16716062 TI - The heart rate of breath-hold divers during static apnea: effects of competitive stress. AB - Breath-hold divers compete with regard to depth, time and/or distance. The present observations were carried out on athletes performing static apnea where they perform one breath-hold for as long a duration as possible with the body and face immersed in water. Heart rate was measured on eight competitors participating in the Swedish Championship in static apnea 2001, both during the competition and during a separate training session using the Polar NV system. The duration of apneas during the competition ranged from 3 minutes 27 seconds to 5 minutes 33 seconds. The divers exhibited significantly faster heart rates prior to and during the first 90 seconds of apnea in connection with competition, than during training. One subject experienced a loss of motor control during the competition. We suggest that mental stress in humans, caused here by a competitive situation, leads to an increase in the heart rate during apnea. PMID- 16716063 TI - Plasma glucose response to recreational diving in novice teenage divers with insulin-requiring diabetes mellitus. AB - A growing number of individuals with insulin-requiring diabetes mellitus (IRDM) dive, but data on plasma glucose (PG) response to diving are limited, particularly for adolescents. We report on seven 16-17 year old novice divers with IRDM participating in a tropical diving camp who had recent at least moderate PG control (HbA1c 7.3 +/- 1.1%) (mean +/- SD). PG was measured at 60, 30 and 10 min pre-dive and immediately following 42 dives. Maximum depth (17 +/- 6 msw) and total underwater times (44 +/- 14 min) were not extreme. Pre-dive PG exceeded 16.7 mmol x L(-1) (300 mg x dL(-1)) in 22% of dives. Males had significantly higher pre-dive levels (15.4 +/- 5.6 mmol x L(-1) [277 +/- 100 mg x dL(-1)] vs. 12.8 +/- 2.9 mmol x L(-1) [230 +/- 52 mg x dL(-1)], respectively) and greater pre-post-dive changes (-4.3 +/- 4.4 mmol x L(-1) [-78 +/- 79 mg x dL(-1)] vs. -0.5 +/- 4.3 mmol x L(-1) [-9 +/- 77 mg x dL(-1)], respectively). Post-dive PG was < 4.4 mmol x L(-1) [< 80 mg x dL(-1)] in two dives by two different males (3.4 and 3.9 mmol x L(-1) [61 and 70 mg x dL(-1)]). No symptoms or complications of hypoglycemia were reported. These data show that in a closely monitored situation, and with benign diving conditions, some diabetic adolescents with good control and no secondary complications may be able to dive safely. The impact of purposeful elevation of PG to protect against hypoglycemia during diving remains to be determined. PMID- 16716064 TI - The absence of hearing loss in otologically asymptomatic recreational scuba divers. AB - We undertook a retrospective cohort study of 16 experienced recreational scuba divers and 16 matched non-diver controls to determine the prevalence of hearing loss and, if present, the likely causes of this loss. Each subject was required to be aged 55 years or less and to have no history or likelihood of hearing loss. An audiologist, blinded to each subject's group status, undertook all examinations. There were no significant differences in group demographics. All divers were highly experienced (median number of dives 725). Comparison of mean hearing thresholds (range 250-8000 Hz) revealed no significant differences between divers and non-divers for both air and bone conduction studies. The only exception was at 6000 Hz where the air conduction threshold was significantly higher in divers than in non-divers (p = 0.03). However, there were no significant differences in Pure Tone and High Frequency averages. We conclude that experienced recreational scuba divers do not have elevated hearing threshold levels overall when compared to non-diver controls. This conclusion differs from that of investigators who have examined the hearing of experienced professional divers. Further investigation is indicated to further investigate this discrepancy and to determine whether the apparent hearing loss among the divers at 6000Hz was an isolated departure from normal hearing thresholds or, in fact, the result of diving. PMID- 16716065 TI - Structural and thermodynamic basis for the enhanced transcriptional control by the human papillomavirus strain-16 E2 protein. AB - Strain 16 of the human papillomavirus is responsible for the largest number of cases of cervical cancers linked to this virus, and the E2 protein is the transcriptional regulator of all viral genes. We present the first structure for the DNA binding domain of HPV16 E2 bound to DNA, and in particular, to a natural cognate sequence. The NMR structure of the protein backbone reveals that the overall conformation remains virtually unchanged, and chemical shift analysis of the protein bound to a shorter DNA duplex uncovered a contact out of the minimal E2 DNA binding site, made by lysine 349. This contact was confirmed by titration calorimetry and mutagenesis, with a contribution of 1.0 kcal mol(-)(1) to binding energy. HPV16 E2 has the highest DNA binding affinity and exerts a strict transcriptional control, translated into the repression of the E6 and E7 oncogenes. These novel features provide the structural and thermodynamic basis for this tight transcriptional control, the loss of which correlates with carcinogenesis. PMID- 16716066 TI - The conformational dynamics of a metastable serpin studied by hydrogen exchange and mass spectrometry. AB - Serpins are a class of protease inhibitors that initially fold to a metastable structure and subsequently undergo a large conformational change to a stable structure when they inhibit their target proteases. How serpins are able to achieve this remarkable conformational rearrangement is still not understood. To address the question of how the dynamic properties of the metastable form may facilitate the conformational change, hydrogen/deuterium exchange and mass spectrometry were employed to probe the conformational dynamics of the serpin human alpha(1)-antitrypsin (alpha(1)AT). It was found that the F helix, which in the crystal structure appears to physically block the conformational change, is highly dynamic in the metastable form. In particular, the C-terminal half of the F helix appears to spend a substantial fraction of time in a partially unfolded state. In contrast, beta-strands 3A and 5A, which must separate to accommodate insertion of the reactive center loop (RCL), are not conformationally flexible in the metastable state but are rigid and stable. The conformational lability required for loop insertion must therefore be triggered during the inhibition reaction. Beta-strand 1C, which anchors the distal end of the RCL and thus prevents transition to the so-called latent form, is also stable, consistent with the observation that alpha(1)AT does not spontaneously adopt the latent form. A surprising degree of flexibility is seen in beta-strand 6A, and it is speculated that this flexibility may deter the formation of edge-edge polymers. PMID- 16716067 TI - Cyan and yellow super fluorescent proteins with improved brightness, protein folding, and FRET Forster radius. AB - Enhanced cyan and yellow fluorescent proteins are widely used for dual color imaging and protein-protein interaction studies based on fluorescence resonance energy transfer. Use of these fluorescent proteins can be limited by their thermosensitivity, dim fluorescence, and tendency for aggregation. Here we report the results of a site-directed mutagenesis approach to improve these fluorescent proteins. We created monomeric optimized variants of ECFP and EYFP, which fold faster and more efficiently at 37 degrees C and have superior solubility and brightness. Bacteria expressing SCFP3A were 9-fold brighter than those expressing ECFP and 1.2-fold brighter than bacteria expressing Cerulean. SCFP3A has an increased quantum yield (0.56) and fluorescence lifetime. Bacteria expressing SYFP2 were 12 times brighter than those expressing EYFP(Q69K) and almost 2-fold brighter than bacteria expressing Venus. In HeLa cells, the improvements were less pronounced; nonetheless, cells expressing SCFP3A and SYFP2 were both 1.5 fold brighter than cells expressing ECFP and EYFP(Q69K), respectively. The enhancements of SCFP3A and SYFP2 are most probably due to an increased intrinsic brightness (1.7-fold and 1.3-fold for purified recombinant proteins, compared to ECFP & EYFP(Q69K), respectively) and due to enhanced protein folding and maturation. The latter enhancements most significantly contribute to the increased fluorescent yield in bacteria whereas they appear less significant for mammalian cell systems. SCFP3A and SYFP2 make a superior donor-acceptor pair for fluorescence resonance energy transfer, because of the high quantum yield and increased lifetime of SCFP3A and the high extinction coefficient of SYFP2. Furthermore, SCFP1, a CFP variant with a short fluorescence lifetime but identical spectra compared to ECFP and SCFP3A, was characterized. Using the large lifetime difference between SCFP1 and SCFP3A enabled us to perform for the first time dual-lifetime imaging of spectrally identical fluorescent species in living cells. PMID- 16716068 TI - Light control of mitochondrial complex I activity by a photoresponsive inhibitor. AB - We recently developed a new class of inhibitors of bovine heart mitochondrial NADH-ubiquinone oxidoreductase (complex I), named Deltalac-acetogenin [Ichimaru et al. (2005) Biochemistry 44, 816-825]. The inhibitory potency of Deltalac acetogenin is remarkably affected by the molecular shape of the alkyl side chains. We speculated that if the shape of the side chains can be changed by the trans-cisphotoisomerization of the azobenzene unit that is introduced into the chain moiety, the inhibitory effect could be switched on and off in a reversible manner. Such a photoresponsive inhibitor may allow rapid, remote, and noninvasive control of complex I activity. Therefore, we here synthesized Deltalac-acetogenin (3) possessing an azobenzene unit in the side chains. (1)H NMR, HPLC, and UV visible absorption analyses indicated that the azobenzene unit in 3 is rapidly and reversibly trans-cis isomerized by photoirradiation in chloroform and ethanol. The inhibitory effect of trans,trans-3 on complex I activity in submitochondrial particles was more potent than that of cis,cis-3. When 3 was applied at the nanomolar level to complex I, the inhibitory effect was reversibly reduced and enhanced by alternating irradiation by UV and visible light, respectively. The present study gives a positive clue to the light control of complex I activity. PMID- 16716069 TI - Reaction geometry and thermostable variant of pyranose 2-oxidase from the white rot fungus Peniophora sp. AB - Pyranose 2-oxidase catalyzes the oxidation of a number of carbohydrates using dioxygen; glucose, for example, is oxidized at carbon 2. The structure of pyranose 2-oxidase with the reaction product 2-keto-beta-d-glucose bound in the active center is reported in a new crystal form at 1.41 A resolution. The binding structure suggests that the alpha-anomer cannot be processed. The binding mode of the oxidized product was used to model other sugars accepted by the enzyme and to explain its specificity and catalytic rates. The reported structure at pH 6.0 shows a drastic conformational change in the loop of residues 454-461 (loop 454 461) at the active center compared to that of a closely homologous enzyme analyzed at pH 4.5 with a bound acetate inhibitor. In our structures, the loop is highly mobile and shifts to make way for the sugar to pass into the active center. Presumably, loop 454-461 functions as a gatekeeper. Apart from the wild type enzyme, a thermostable variant was analyzed at 1.84 A resolution. In this variant, Glu542 is exchanged for a lysine. The observed stabilization could be a result of the mutated residue changing an ionic contact at a comparatively weak interface of the tetramer. PMID- 16716070 TI - Effects of temperature and viscosity on R67 dihydrofolate reductase catalysis. AB - R67 dihydrofolate reductase (DHFR) is a novel homotetrameric protein that possesses 222 symmetry and a single, voluminous active site pore. This symmetry poses numerous limitations on catalysis; for example, two dihydrofolate (DHF) molecules or two NADPH molecules, or one substrate plus one cofactor can bind. Only the latter combination leads to catalysis. To garner additional information on how this enzyme facilitates transition-state formation, the temperature dependence of binding and catalysis was monitored. The binding of NADPH and DHF is enthalpy-driven. Previous primary isotope effect studies indicate hydride transfer is at least partially rate-determining. Accordingly, the activation energy associated with transition-state formation was measured and is found to be 6.9 kcal/mol (DeltaH(++)(25) = 6.3 kcal/mol). A large entropic component is also found associated with catalysis, TDeltaS(++)(25) = -11.3 kcal/mol. The poor substrate, dihydropteroate, binds more weakly than dihydrofolate (DeltaDeltaG = 1.4 kcal/mol) and displays a large loss in the binding enthalpy value (DeltaDeltaH = 3.8 kcal/mol). The k(cat) value for dihydropteroate reduction is decreased 1600-fold compared to DHF usage. This effect appears to derive mostly from the DeltaDeltaH difference in binding, demonstrating that the glutamate tail is important for catalysis. This result is surprising, as the para-aminobenzoyl glutamate tail of DHF has been previously shown to be disordered by both NMR and crystallography studies. Viscosity studies were also performed and confirmed that the hydride transfer rate is not sensitive to sucrose addition. Surprisingly, binding of DHF, by both K(m) and K(d) determination, was found to be sensitive to added viscogens, suggesting a role for water in DHF binding. PMID- 16716071 TI - Closing of the flaps of HIV-1 protease induced by substrate binding: a model of a flap closing mechanism in retroviral aspartic proteases. AB - The active site of aspartic proteases is covered by one or more flaps, which control access to the active site and play a significant role in the binding of the substrate. An extensive conformational change of the flaps takes place upon binding of substrate to the active site. A long molecular dynamics simulation was performed on the complex consisting of a peptide (CA-p2) from a natural substrate cleavage site of the gag/pol polyprotein placed in the active site of HIV-1 protease (PR) with an open flap conformation. During the simulation, the substrate induced the closing of the flaps into the closed conformation in an asymmetrical way through a hydrophobic intermediate state cluster. The nature of the residues of HIV-1 PR identified to be important in the flap closing mechanism is conserved across known structures of retroviral aspartic proteases family. The flap closing mechanism described in HIV-1 PR is proposed to be a general model for flap closing in retroviral aspartic proteases. PMID- 16716072 TI - The extraordinary specificity of xanthine phosphoribosyltransferase from Bacillus subtilis elucidated by reaction kinetics, ligand binding, and crystallography. AB - Xanthine phosphoribosyltransferase (XPRTase) from Bacillus subtilis is a representative of the highly xanthine specific XPRTases found in Gram-positive bacteria. These XPRTases constitute a distinct subclass of 6-oxopurine PRTases, which deviate strongly from the major class of H(X)GPRTases with respect to sequence, PRPP binding motif, and oligomeric structure. They are more related with the PurR repressor of Gram-positive bacteria, the adenine PRTase, and orotate PRTase. The catalytic function and high specificity for xanthine of B. subtilis XPRTase were investigated by ligand binding studies and reaction kinetics as a function of pH with xanthine, hypoxanthine, and guanine as substrates. The crystal structure of the dimeric XPRTase-GMP complex was determined to 2.05 A resolution. In a sequential reaction mechanism XPRTase binds first PRPP, stabilizing its active dimeric form, and subsequently xanthine. The XPRTase is able also to react with guanine and hypoxanthine albeit at much lower (10(-)(4)-fold) catalytic efficiency. Different pK(a) values for the bases and variations in their electrostatic potential can account for these catalytic differences. The unique base specificity of XPRTase has been related to a few key residues in the active site. Asn27 can in different orientations form hydrogen bonds to an amino group or an oxo group at the 2-position of the purine base, and Lys156 is positioned to make a hydrogen bond with N7. This and the absence of a catalytic carboxylate group near the N7-position require the purine base to dissociate a proton spontaneously in order to undergo catalysis. PMID- 16716073 TI - Alpha-amino-beta-carboxymuconic-epsilon-semialdehyde decarboxylase (ACMSD) is a new member of the amidohydrolase superfamily. AB - The enzymatic activity of Pseudomonas fluorescens alpha-amino-beta-carboxymuconic epsilon-semialdehyde decarboxylase (ACMSD) is critically dependent on a transition metal ion [Li, T., Walker, A. L., Iwaki, H., Hasegawa, Y., and Liu, A. (2005) J. Am. Chem. Soc. 127, 12282-12290]. Sequence analysis in this study further suggests that ACMSD belongs to the amidohydrolase superfamily, whose structurally characterized members comprise a catalytically essential metal cofactor. To identify ACMSD's metal ligands and assess their functions in catalysis, a site-directed mutagenesis analysis was conducted. Alteration of His 9, His-177, and Asp-294 resulted in a dramatic loss of enzyme activity, substantial reduction of the metal-binding ability, and an altered metallocenter electronic structure. Thus, these residues are confirmed to be the endogenous metal ligands. His-11 is implicated in metal binding because of the strictly conserved HxH motif with His-9. Mutations at the 228 site yielded nearly inactive enzyme variants H228A and H228E. The two His-228 mutant proteins, however, exhibited full metal-binding ability and a metal center similar to that of the wild-type enzyme as shown by EPR spectroscopy. Kinetic analysis on the mutants indicates that His-228 is a critical catalytic residue along with the metal cofactor. Since the identified metal ligands and His-228 are present in all known ACMSD sequences, it is likely that ACMSD proteins from other organisms contain the same cofactor and share similar catalytic mechanisms. ACMSD is therefore the first characterized member in the amidohydrolase superfamily that represents a C C breaking activity. PMID- 16716074 TI - Optimization of a ribosomal structural domain by natural selection. AB - A conserved, independently folding domain in the large ribosomal subunit consists of 58 nt of rRNA and a single protein, L11. The tertiary structure of an rRNA fragment carrying the Escherichia coli sequence is marginally stable in vitro but can be substantially stabilized by mutations found in other organisms. To distinguish between possible reasons why natural selection has not evolved a more stable rRNA structure in E. coli, mutations affecting the rRNA tertiary structure were assessed for their in vitro effects on rRNA stability and L11 affinity (in the context of an rRNA fragment) or in vivo effects on cell growth rate and L11 content of ribosomes. The rRNA fragment stabilities ranged from -4 to +9 kcal/mol relative to the wild-type sequence. Variants in the range of -4 to +5 kcal/mol had almost no observable effect in vivo, while more destabilizing mutations (>7 kcal/mol) were not tolerated. The data suggest that the in vivo stability of the complex is roughly -6 kcal/mol and that any single tertiary interaction is dispensable for function as long as a minimum stability of the complex is maintained. On the basis of these data, it seems that the evolution of this domain has not been constrained by inherent structural or functional limits on stability. The estimated stability corresponds to only a few ribosomes per bacterial cell dissociated from L11 at any time; thus the selective advantage for any further increase in stability may be so small as to be outweighed by other competing selective pressures. PMID- 16716076 TI - Regulation of integrin alphaIIbbeta3 activation by distinct regions of its cytoplasmic tails. AB - The short cytoplasmic tails regulate activation of integrin adhesion receptors via clasping/unclasping of their membrane-proximal helices. Using integrin alpha(IIb)beta(3) as a model, we show that a previously reported activating mutation alpha(IIb)(R(995)D) that perturbs the electrostatic interface in the clasp only partially activates alpha(IIb)beta(3) and that extensive activation of the receptor is achieved by complete deletion of alpha(IIb) CT or triple mutations in alpha(IIb)(V(990)A/F(992)A/R(995)D) that disrupt both electrostatic and hydrophobic interfaces in the clasp. The results provide quantitative evidence for an equilibrium-based integrin activation process where shifting the equilibrium to the fully activated state requires total unclasping of the cytoplasmic tails. We further demonstrate that while the C-terminal region of the alpha(IIb) tail minimally influences alpha(IIb)beta(3) activation, the C-terminal region of the beta(3) tail is critically involved. A disease-causing mutation of S(752)P in this region, but not S(752)A, suppressed partial activation induced by R(995)D or the talin head domain but did not affect activation induced by alpha(IIb) truncation. NMR spectroscopy revealed that S(752)P but not the S(752)A mutation disrupted a C-terminal helix within the beta(3) tail, suggesting that the C-terminal helix may regulate the equilibrium-based clasping/unclasping process. Together, these data provide molecular insights into how distinct regions of the cytoplasmic tails differentially and cooperatively regulate integrin activation. PMID- 16716075 TI - Flexible 5-guanidino-4-nitroimidazole DNA lesions: structures and thermodynamics. AB - 5-Guanidino-4-nitroimidazole (NI), derived from guanine oxidation by reactive oxygen and nitrogen species, contains an unusual flexible ring-opened structure, with nitro and guanidino groups which possess multiple hydrogen bonding capabilities. In vitro primer extension experiments with bacterial and mammalian polymerases show that NI incorporates C as well as A and G opposite the lesion, depending on the polymerase. To elucidate structural and thermodynamic properties of the mutagenic NI lesion, we have investigated the structure of the modified base itself and the NI-containing nucleoside with high-level quantum mechanical calculations and have employed molecular modeling and molecular dynamics simulations in solution for the lesion in B-DNA duplexes, with four partner bases opposite the NI. Our results show that NI adopts a planar structure at the damaged base level. However, in the nucleoside and in DNA duplexes, steric hindrance between the guanidino group and its linked sugar causes NI to be nonplanar. The NI lesion can adopt both syn and anti conformations on the DNA duplex level, with the guanidino group positioned in the DNA major and minor grooves, respectively; the specific preference depends on the partner base. On the basis of hydrogen bonding and stacking interactions, groove dimensions, and bending, we find that the least distorted NI-modified duplex contains partner C, consistent with observed incorporation of C opposite NI. However, hydrogen bonding interactions between NI and partner G or A are also found, which would be compatible with the observed mismatches. PMID- 16716077 TI - Effect of glycosylation on the function of a soluble, recombinant form of the transferrin receptor. AB - Production of the soluble portion of the transferrin receptor (sTFR) by baby hamster kidney (BHK) cells is described, and the effect of glycosylation on the biological function of sTFR is evaluated for the first time. The sTFR (residues 121-760) has three N-linked glycosylation sites (Asn251, Asn317, and Asn727). Although fully glycosylated sTFR is secreted into the tissue culture medium ( approximately 40 mg/L), no nonglycosylated sTFR could be produced, suggesting that carbohydrate is critical to the folding, stability, and/or secretion of the receptor. Mutants in which glycosylation at positions 251 and 727 (N251D and N727D) is eliminated are well expressed, whereas production of the N317D mutant is poor. Analysis by electrospray ionization mass spectrometry confirms dimerization of the sTFR and the absence of the carbohydrate at the single site in each mutant. The effect of glycosylation on binding to diferric human transferrin (Fe(2) hTF), an authentic monoferric hTF with iron in the C-lobe (designated Fe(C) hTF), and a mutant (designated Mut-Fe(C) hTF that features a 30 fold slower iron release rate) was determined by surface plasmon resonance; a small ( approximately 20%) but consistent difference is noted for the binding of Fe(C) hTF and the Mut-Fe(C) hTF to the sTFR N317D mutant. The rate of iron release from Fe(C) hTF and Mut-Fe(C) hTF in complex with the sTFR and the sTFR mutants at pH 5.6 reveals that only the N317D mutant has a significant effect. The carbohydrate at position 317 lies close to a region of the TFR previously shown to interact with hTF. PMID- 16716078 TI - Effect of copper and manganese on the de novo generation of protease-resistant prion protein in yeast cells. AB - The prion protein (PrP) is the key protein implicated in diseases known as transmissible spongiform encephalopathies. PrP has been shown to bind manganese and copper, the latter being involved in the normal function of the protein. Indeed, upon expression in yeast we noted a major increase in intracellular copper and a decrease in manganese. Interestingly, protease-resistant PrP(Sc) like protein (PrP(res)) formation was induced when PrP-expressing yeast cells were grown in copper- and/or manganese-supplemented media. The pattern of PrP banding in SDS-PAGE was dominantly determined by manganese. This conformational transition was stable against EDTA treatment but not in the presence of the copper chelators bathocuproinedisulfonic acid or clioquinol. Conclusively, PrP itself influences manganese and copper metabolism, and a replacement of copper in PrP complexes with manganese is highly likely under the condition of copper depletion or if excess amounts of copper and manganese are present. Taken together, our present study demonstrates the involvement of PrP in the regulation of intracellular metal ion homeostasis and uncovers copper and, more severely, manganese ions as in vivo risk factors for the conversion into PrP(Sc). PMID- 16716079 TI - A spectroscopic study on the interactions of porphyrin with G-quadruplex DNAs. AB - Free-base porphyrin (5,10,15,20-tetrakis(1-methyl-4-pyridyl)-21H,23H-porphine) (H(2)TMPyP4) has been shown to be an effective telomerase inhibitor by an in vitro assay. Here, we examined the interactions of the H(2)TMPyP4 with three distinct G-quadruplex DNAs, the parallel-stranded (TG(4)T)4, dimer-hairpin-folded (G(4)T(4)G(4))2, and monomer-folded AG(3)(T(2)AG(3))(3), by ultraviolet resonance Raman spectroscopy (UVRR), UV-vis absorption spectroscopy, fluorescence spectroscopy, and surface-enhanced Raman spectroscopy (SERS). The data obtained by the continuous variation titration method show that the binding stoichiometry of H(2)TMPyP4/G-quadruplex is 2:1 for (TG(4)T)4 and 4:1 for (G(4)T(4)G(4))2 or AG(3)(T(2)AG(3))(3). The results of SERS spectra, UV-vis absorption titration, and fluorescence emission spectra together with the binding stoichiometries reveal that two H(2)TMPyP4 molecules are externally stacked at two ends of the parallel (TG(4)T)4 G-quadruplex, whereas H(2)TMPyP4 molecules can intercalate within their diagonal or lateral loop regions and intervals between two G-tetrads for (G(4)T(4)G(4))2 and AG(3)(T(2)AG(3))(3) G-quadruplexes. The binding of H(2)TMPyP4 to (TG(4)T)4 G-quadruplex results in the hypochromicity of the UV Raman signal of (TG(4)T)4, indicating that the stacking effects between H(2)TMPyP4 and DNA bases are significant. The Raman hyperchromicities and shifts are observed after the binding of H(2)TMPyP4 to both (G(4)T(4)G(4))2 and AG(3)(T(2)AG(3))(3) G-quadruplexes. This indicates that the intercalative H(2)TMPyP4 can lengthen the vertical distance between adjacent G-tetrads of (G(4)T(4)G(4))2 and AG(3)(T(2)AG(3))(3) and change their conformations. The present study provides new insights into the effect of H(2)TMPyP4 binding on the structures of G-quadruplexes and also demonstrates that Raman spectroscopy is an ideal method for examining the interaction between drugs and G-quadruplexes. PMID- 16716080 TI - Synthesis of HPr(Ser-P)(His-P) by enzyme I of the phosphoenolpyruvate: sugar phosphotransferase system of Streptococcus salivarius. AB - HPr is a protein of the bacterial phosphoenolpyruvate:sugar phosphotransferase transport system (PTS). In Gram-positive bacteria, HPr can be phosphorylated on Ser(46) by HPr(Ser) kinase/phosphorylase (HPrK/P) and on His(15) by enzyme I (EI) of the PTS. In vitro studies have shown that phosphorylation on one residue greatly inhibits the second phosphorylation. However, streptococci contain significant amounts of HPr(Ser-P)(His approximately P) during exponential growth, and recent studies suggest that phosphorylation of HPr(Ser-P) by EI is involved in the recycling of HPr(Ser-P)(His approximately P). We report in this paper a study on the phosphorylation of Streptococcus salivarius HPr, HPr(Ser-P), and HPr(S46D) by EI. Our results indicate that (i) the specificity constant (k(cat)/K(m)) of EI for HPr(Ser-P) at pH 7.9 was approximately 5000-fold smaller than that observed for HPr, (ii) no metabolic intermediates were able to stimulate HPr(Ser-P) phosphorylation, (iii) the rate of HPr phosphorylation decreased at pHs below 6.5, while that of HPr(Ser-P) increased and was almost 10 fold higher at pH 6.1 than at pH 7.9, (iv) HPr(S46D), a mutated HPr alleged to mimic HPr(Ser-P), was also phosphorylated more efficiently under acidic conditions, and, lastly, (v) phosphorylation of Bacillus subtilis HPr(Ser-P) by B. subtilis EI was also stimulated at acidic pH. Our results suggest that the high levels of HPr(Ser-P)(His approximately P) in streptococci result from the combination of two factors, a high physiological concentration of HPr(Ser-P) and stimulation of HPr(Ser-P) phosphorylation by EI at acidic pH, an intracellular condition that occurs in response to the acidification of the external medium during growth of the culture. PMID- 16716081 TI - Heparin activates beta-secretase (BACE1) of Alzheimer's disease and increases autocatalysis of the enzyme. AB - BACE1 is an aspartic protease that generates the N-terminus of the beta-amyloid protein (Alphabeta) from the beta-amyloid precursor protein (APP). BACE1 is a key target for Alzheimer drug development. However, little is known about the physiological regulation of the enzyme. Heparin can promote beta-secretase cleavage of APP in neuroblastoma cells. However, heparin has also been reported to directly inhibit BACE1 activity in vitro. To clarify the role of heparin in regulating BACE1, we examined the effect of heparin on the activity of recombinant human BACE1 (rBACE1) in vitro. Low concentrations (1 microg/mL) of heparin were found to stimulate rBACE1, increasing enzyme V(max) and decreasing the K(M). In contrast, higher concentrations of heparin (10 or 100 microg/mL) were inhibitory. Heparin affinity chromatography demonstrated that heparin interacted strongly with the zymogen form of rBACE1 and bound to a peptide homologous to the N-terminal pro sequence of BACE1. Mature (pro sequence cleaved) enzyme lacked the capacity to be stimulated by heparin, indicating that the pro domain was necessary for the stimulation by heparin. Furthermore, in the presence of stimulatory concentrations of heparin, there was an increase in autocatalytic cleavage of the protease domain and a subsequent loss of enzyme activity in vitro. Our results strongly suggest that heparin stimulates the partially active BACE1 zymogen, and we propose that the activation is mediated by high-affinity binding of heparin to the pro domain. Our study provides evidence that heparan sulfate proteoglycans could regulate the rate of Alphabeta production in vivo. PMID- 16716082 TI - The peripheral stalk participates in the yeast ATP synthase dimerization independently of e and g subunits. AB - It is now clearly established that dimerization of the F(1)F(o) ATP synthase takes place in the mitochondrial inner membrane. Interestingly, oligomerization of this enzyme seems to be involved in cristae morphogenesis. As they were able to form homodimers, subunits 4, e, and g have been proposed as potential ATP synthase dimerization subunits. In this paper, we provide evidence that subunit h, a peripheral stalk component, is located either at or near the ATP synthase dimerization interface. Subunit h homodimers were formed in mitochondria and were found to be associated to ATP synthase dimers. Moreover, homodimerization of subunit h and of subunit i turned out to be independent of subunits e and g, confirming the existence of an ATP synthase dimer in the mitochondrial inner membrane in the absence of subunits e and g. For the first time, this dimer has been observed by BN-PAGE. Finally, from these results we are now able to update our model for the supramolecular organization of the ATP synthase in the membrane and propose a role for subunits e and g, which stabilize the ATP synthase dimers and are involved in the oligomerization of the complex. PMID- 16716083 TI - Aggregation/fibrillogenesis of recombinant human prion protein and Gerstmann Straussler-Scheinker disease peptides in the presence of metal ions. AB - In this study we investigated the role of Cu(2+), Mn(2+), Zn(2+), and Al(3+) in inducing defective conformational rearrangements of the recombinant human prion protein (hPrP), which trigger aggregation and fibrillogenesis. The research was extended to the fragment of hPrP spanning residues 82-146, which was identified as a major component of the amyloid deposits in the brain of patients affected by Gerstmann-Straussler-Scheinker (GSS) disease. Variants of the 82-146 wild-type subunit [PrP-(82-146)(wt)] were also examined, including entirely, [PrP-(82 146)(scr)], and partially scrambled, [PrP-(82-146)(106)(-)(126scr)] and [PrP-(82 146)(127)(-)(146scr)], peptides. Al(3+) strongly stimulated the conversion of native hPrP into the altered conformation, and its potency in inducing aggregation was very high. Despite a lower rate and extent of prion protein conversion into altered isoforms, however, Zn(2+) was more efficient than Al(3+) in promoting organization of hPrP aggregates into well-structured, amyloid-like fibrillar filaments, whereas Mn(2+) delayed and Cu(2+) prevented the process. GSS peptides underwent the fibrillogenesis process much faster than the full-length protein. The intrinsic ability of PrP-(82-146)(wt) to form fibrillar aggregates was exalted in the presence of Zn(2+) and, to a lesser extent, of Al(3+), whereas Cu(2+) and Mn(2+) inhibited the conversion of the peptide into amyloid fibrils. Amino acid substitution in the neurotoxic core (sequence 106-126) of the 82-146 fragment reduced its amyloidogenic potential. In this case, the stimulatory effect of Zn(2+) was lower as compared to the wild-type peptide; on the contrary Al(3+) and Mn(2+) induced a higher propensity to fibrillation, which was ascribed to different binding modalities to GSS peptides. In all cases, alteration of the 127-146 sequence strongly inhibited the fibrillogenesis process, thus suggesting that integrity of the C-terminal region was essential both to confer amyloidogenic properties on GSS peptides and to activate the stimulatory potential of the metal ions. PMID- 16716084 TI - 14-3-3 Mediates phosphorylation-dependent inhibition of the interaction between the ubiquitin E3 ligase Nedd4-2 and epithelial Na+ channels. AB - Although recent studies show that the 14-3-3 protein is a negative regulator of ubiquitin E3 protein ligases, the molecular mechanism remains largely unknown. We previously demonstrated that 14-3-3 specifically binds one of the E3 enzymes, Nedd4-2 (a human gene product of KIAA0439, termed hNedd4-2), which can be phosphorylated by serum glucocorticoid-inducible protein kinase 1 (SGK1); this binding protects the phosphorylated/inactive hNedd4-2 from phosphatase-catalyzed dephosphorylation [Ichimura, T., et al. (2005) J. Biol. Chem. 280, 13187-13194]. Here we report an additional mechanism of 14-3-3-mediated regulation of hNedd4-2. Using surface plasmon resonance spectrometry, we show that 14-3-3 inhibits the interaction between the WW domains of hNedd4-2 and the PY motif of the epithelial Na(+) channel, ENaC. The inhibition was dose-dependent and was dependent on SGK1 catalyzed phosphorylation of Ser468 located between the WW domains. Importantly, a mutant of hNedd4-2, which can be phosphorylated by SGK1 but cannot bind 14-3-3, reduced SGK1-mediated stimulation of the ENaC-induced current in Xenopus laevis oocytes. In addition, 14-3-3 had similar effects on hNedd4-2 that had been phosphorylated by cAMP-dependent protein kinase (PKA). Our results, together with the recent finding on 14-3-3/parkin interactions [Sato, S., et al. (2006) EMBO J. 25, 211-221], suggest that 14-3-3 suppresses ubiquitin E3 ligase activities by inhibiting the formation of the enzyme/substrate complex. PMID- 16716086 TI - Human polynucleotide phosphorylase protein in response to oxidative stress. AB - 8-Oxo-7,8-dihydroguanine (8-oxoGua) is generated in nucleic acids as well as in their precursors due to the actions of oxygen radicals produced through a normal cellular metabolism. Since oxidized guanine can pair with both cytosine and adenine, it causes alterations in the phenotypic expression when it is present in RNA. To prevent such an outcome, organisms must have some mechanism for eliminating such oxidized guanine nucleotides from RNA and its precursors. In mammalian cells, MTH1 and NUDT5 proteins degrade 8-oxoGTP and 8-oxoGDP to 8 oxoGMP, which is an unusable form for RNA synthesis. In a search for proteins functioning at the RNA level, polynucleotide phosphorylase (PNP) protein has been suggested to be a good candidate for such a role. The human PNP protein has an ability to bind specifically to RNA containing 8-oxoGua. When human cells are exposed to agents that induce oxidative stress, such as hydrogen peroxide and menadion, the amounts of PNP protein decrease rapidly while amounts of other proteins in the cells do not change after such treatments. No specific decrease in the PNP protein level is observed when cells are treated with ACNU and cycloheximide at doses sufficient to provide the same degree of growth suppression. These results imply that the PNP protein might thus play a role in excluding oxidized forms of RNA from the translation mechanism. PMID- 16716085 TI - The interaction of recombinant subdomains of the procollagen C-proteinase with procollagen I provides a quantitative explanation for functional differences between the two splice variants, mammalian tolloid and bone morphogenetic protein 1. AB - The procollagen C-proteinase (PCP) is a zinc peptidase of the astacin family and the metzincin superfamily. The enzyme removes the C-terminal propeptides of fibrillar procollagens and activates other matrix proteins. Besides its catalytic protease domain, the procollagen C-proteinase contains several C-terminal CUB modules (named after complement factors C1r and C1s, the sea urchin UEGF protein, and BMP-1) and EGF-like domains. The two major splice forms of the C-proteinase differ in their overall domain composition. The longer variant, termed mammalian tolloid (mTld, i.e., PCP-2), has the protease-CUB1-CUB2-EGF1-CUB3-EGF2-CUB4-CUB5 composition, whereas the shorter form termed bone morphogenetic protein 1 (BMP-1, i.e., PCP-1) ends after the CUB3 domain. Two related genes encode proteases similar to mTld in humans and have been termed mammalian tolloid like-1 and -2 (mTll-1 and mTll-2, respectively). For mTll-1, it has been shown that it has C proteinase activity. We demonstrate that recombinant EGF1-CUB3, CUB3, CUB3-EGF2, EGF2-CUB4, and CUB4-CUB5 modules of the procollagen C-proteinase can be expressed in bacteria and adopt a functional antiparallel beta-sheet conformation. As shown by surface plasmon resonance analysis, the modules bind to procollagen I in a 1:1 stoichiometry with dissociation constants (K(D)) ranging from 622.0 to 1.0 nM. Their binding to mature collagen I is weaker by at least 1 order of magnitude. Constructs containing EGF domains bind more strongly than those consisting of CUB domains only. This suggests that a combination of CUB and EGF domains serves as the minimal functional unit. The binding affinities of the EGF-containing modules for procollagen increase in the order EGF1-CUB3 < CUB3-EGF2 < EGF2-CUB4. In the context of the full length PCP, this implies that a given module has an affinity that continues to increase the more C-terminally the module is located within the PCP. The tightest binding module, EGF2-CUB4 (K(D) = 1.0 nM), is only present in mTld, which might provide a quantitative explanation for the different efficiencies of BMP-1 and mTld in procollagen C-proteinase activity. PMID- 16716087 TI - Identification of FX in the heliobacterial reaction center as a [4Fe-4S] cluster with an S = 3/2 ground spin state. AB - Type I homodimeric reaction centers, particularly the class present in heliobacteria, are not well understood. Even though the primary amino acid sequence of PshA in Heliobacillus mobilis has been shown to contain an F(X) binding site, a functional Fe-S cluster has not been detected by EPR spectroscopy. Recently, we reported that PshB, which contains F(A)- and F(B)-like Fe-S clusters, could be removed from the Heliobacterium modesticaldum reaction center (HbRC), resulting in 15 ms lifetime charge recombination between P798(+) and an unidentified electron acceptor [Heinnickel, M., Shen, G., Agalarov, R., and Golbeck, J. H. (2005) Biochemistry 44, 9950-9960]. We report here that when a HbRC core is incubated with sodium dithionite in the presence of light, the 15 ms charge recombination is replaced with a kinetic transient in the sub-microsecond time domain, consistent with the reduction of this electron acceptor. Concomitantly, a broad and intense EPR signal arises around g = 5 along with a minor set of resonances around g = 2 similar to the spectrum of the [4Fe-4S](+) cluster in the Fe protein of Azotobacter vinelandii nitrogenase, which exists in two conformations having S = (3)/(2) and S = (1)/(2) ground spin states. The Mossbauer spectrum in the as-isolated HbRC core shows that all of the Fe is present in the form of a [4Fe-4S](2+) cluster. After reduction with sodium dithionite in the presence of light, approximately 65% of the Fe appears in the form of a [4Fe-4S](+) cluster; the remainder is in the [4Fe-4S](2+) state. Analysis of the non-heme iron content of HbRC cores indicates an antenna size of 21.6 +/- 1.1 BChl g molecules/P798. The evidence indicates that the HbRC contains a [4Fe-4S] cluster identified as F(X) that is coordinated between the PshA homodimer; in contrast to F(X) in other type I reaction centers, this [4Fe-4S] cluster exhibits an S = (3)/(2) ground spin state. PMID- 16716089 TI - The status and future directions of cardiac surgery. PMID- 16716090 TI - Colesevelam hydrochloride in the management of dyslipidemia. AB - Dyslipidemia is a highly heterogeneous group of disorders strongly influenced by both genetic and environmental factors. Dyslipidemia significantly increases risk for atherosclerotic disease and all of its various clinical manifestations. Identifying patients with dyslipidemia and initiating therapies aimed at normalizing the lipid profile has been demonstrated to significantly reduce the risk for myocardial infarction, stroke and cardiovascular mortality in both the primary and secondary prevention settings. Guidelines in Europe, Canada and the USA emphasize the need to reduce the burden of atherogenic lipoproteins in serum and to raise levels of high-density lipoproteins in patients at risk for cardiovascular events. Statins have emerged as front-line therapy for managing dyslipidemia, especially in patients with elevated serum levels of low-density lipoprotein cholesterol. As guidelines emphasize the need to reduce serum low density lipoprotein cholesterol to lower levels, goal attainment can be challenging. The use of combination therapy increases the likelihood of therapeutic success for many patients. Furthermore, a significant percentage of patients with dyslipidemia either cannot achieve goals on statin monotherapy, choose not to take a statin or do not tolerate these drugs due to adverse side effects, such as myalgias, weakness or hepatotoxicity. This article summarizes the pharmacology, clinical efficacy and safety of colesevelam hydrochloride, a bile acid-binding resin. Bile acid-binding resins are orally administered anion exchange resins that are not absorbed systemically. These agents bind bile acids and reduce their reabsorption at the level of the terminal ileum and prevent their enterohepatic recirculation. Colesevelam has a favorable side effect and toxicity profile and significantly impacts serum levels of lipoproteins when used as monotherapy or when used in combination with either statins or ezetimibe. PMID- 16716091 TI - Sildenafil, a PDE5 inhibitor, in the treatment of pulmonary hypertension. AB - Pulmonary hypertension is a devastating disorder, characterized by vascular proliferation, intimal hypertrophy and vasoconstriction. In this disorder, alterations in the nitric oxide pathway have borne out to be important in not only vascular proliferation, but also in the maintenance of vascular tone. After synthesis by soluble guanylate cyclase, cGMP effects vasodilation via protein kinase G and other mediators, and is hydrolyzed by phosphodiesterases (PDEs). PDE5 is abundantly expressed in the mammalian lung and its inhibition by sildenafil has been demonstrated to improve pulmonary vascular physiology in vitro and in vivo animal models of pulmonary hypertension. Recent human data has confirmed the efficacy of sildenafil in therapy for humans with pulmonary arterial hypertension. The following review will discuss the underlying basic science supporting the use of sildenafil, as well as human evidence supporting the critical role of this drug in therapy of patients with pulmonary hypertension. PMID- 16716092 TI - Alteplase for acute ischemic stroke. AB - Alteplase, an intravenously administered form of recombinant tissue plasminogen activator (rt-PA), remains the only US FDA-approved thrombolytic treatment for acute ischemic stroke within 3 h of symptom onset. Patients treated with intravenous rt-PA are at least 30% more likely to have minimal or no disability at 3 months compared with placebo. Despite an increased risk of symptomatic intracranial hemorrhage, rt-PA does not increase mortality. The benefit achieved with rt-PA is cost effective and sustained 1 year after treatment. Despite its clear benefit, rt-PA remains underutilized. Although the future of acute ischemic stroke treatment will most likely involve a multi-faceted treatment approach, the primary objective remains to establish recanalization of the involved vessel. For patients with acute ischemic stroke within the first 3 h of symptom onset, rt-PA remains the first step in accomplishing this goal. PMID- 16716093 TI - Perindopril/indapamide combination in the first-line treatment of hypertension and end-organ protection. AB - This article examines evidence-based findings in the literature on the efficacy of perindopril 2 mg/indapamide 0.625 mg, a first-line, low-dose antihypertensive drug combination. In regulatory Phase II and III trials, perindopril/indapamide significantly lowered blood pressure compared with other first-line therapies (atenolol, losartan and irbesartan). This was also the case in STRAtegies of Treatment in Hypertension: Evaluation, a postregistration study versus current monotherapies and stepped-care therapy with different classes of antihypertensive agents. The efficacy/safety ratio (both clinical and with regard to laboratory parameters) of perindopril/indapamide was good. Perindopril/indapamide provides additional antihypertensive efficacy compared with each component used alone and with current monotherapies, with major efficacy on systolic blood pressure, an important predictor of cardiovascular risk. It also reduces pulse pressure, an independent cardiovascular risk factor, large-vessel arterial stiffness and microcirculatory alterations. The fixed dosage of a once-daily tablet, ensures optimal ease of use and enhances patient compliance. Perindopril/indapamide also reduces target organ damage in patients at high cardiovascular risk, such as patients with cardiac hypertrophy and Type 2 diabetics with albuminuria. These benefits, together with the good efficacy/tolerability ratio, fulfill the requirements of the European Society of Hypertension and of the Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure guidelines for low-dose, first-line combination therapy in hypertension. PMID- 16716094 TI - Effects of prematurity on heart rate control: implications for sudden infant death syndrome. AB - In Western countries, 5-11% of all infants are born before 37 weeks of gestation, and with improvements in modern intensive care techniques the number of these preterm infants that survive continues to increase. Preterm birth is one of the leading causes of neonatal morbidity and mortality in developed countries, accounting for 60-80% of infant deaths in those without congenital anomalies. Furthermore, in the post-neonatal period, preterm infants are at four-times greater risk of sudden infant death syndrome. It has been suggested that this increased risk is due to immature autonomic control. This article provides an overview of studies assessing autonomic control of the cardiovascular system in preterm infants. PMID- 16716095 TI - Premature cardiovascular disease in young women with heterozygous familial hypercholesterolemia. AB - Heterozygous familial hypercholesterolemia is associated with elevated low density lipoprotein cholesterol levels and the development of premature cardiovascular disease. Despite this general statement, data regarding the incidence of cardiovascular disease in young women with familial hypercholesterolemia are lacking. In this review, information of age-specific incidence, risk factors and therapeutic avenues in women with heterozygous familial hypercholesterolemia are discussed. PMID- 16716096 TI - Aldosterone antagonists: effective add-on therapy for the treatment of resistant hypertension. AB - Resistant hypertension is defined as blood pressure that remains above target levels despite treatment with three different antihypertensive agents. Cross sectional analyses and hypertension outcome studies indicate that it is a common clinical problem, which will undoubtedly become increasingly prevalent with an aging and increasingly overweight population. Secondary causes of hypertension are common in patients with resistant hypertension, particularly hyperaldosteronism, with a prevalence of approximately 15-20%. This, however, is likely to be an underestimation of the role excess aldosterone plays in causing resistance to treatment. In subjects with resistant hypertension, suppressed renin levels are common, exceeding 60% in studies conducted by the authors and from centers elsewhere in the world, suggesting occurrence of excess aldosterone beyond cases of true primary aldosteronism. Recent clinical studies indicate that aldosterone antagonists provide significant additional blood pressure reduction when added to treatment regimens of patients with resistant hypertension independent of aldosterone levels. These agents are generally well tolerated. Hyperkalemia is an uncommon complication of aldosterone antagonists, but it can occur. Therefore, biochemical monitoring is necessary, particularly in high-risk patients. PMID- 16716097 TI - Management and therapeutic options in pediatric pulmonary hypertension. AB - Idiopathic pulmonary arterial hypertension is a rare and potentially fatal condition. Without treatment, survival is only approximately 2.8 years from diagnosis. However, if the pulmonary hypertension is secondary to other causes, especially to congenital heart disease, it is possible to survive for 30 years or more without treatment. In recent years, remarkable progress has been made, risk factors have been identified and improved imaging techniques, including echocardiography, computer tomography and magnetic resonance imaging, are available. The condition can affect children at any age from fetal life through to adulthood. Patients can present to the respiratory pediatrician with unresponsive asthma, to the neurologist with faints or to the general pediatrician with failure to thrive. Over the last few years there have been significant developments in the available therapy for managing this complicated disease, which have improved the prognostic outlook, such as oral bosentan and sildenafil, intravenous epoprostenol and interventional catheterization with atrial septostomy. This article reviews the current knowledge about causation, investigation and treatment of children with pulmonary hypertension in the clinical setting. PMID- 16716098 TI - Myocardial regeneration by embryonic stem cell transplantation: present and future trends. AB - Embryonic stem cells are a promising source for myocardial regeneration due to their pluripotency and plasticity. In theory, embryonic stem cells are capable of self-renewal in an unlimited fashion, and can differentiate into any cell type required for cell-based therapy, including cardiac myocytes. In recent years, embryonic stem cells have been transplanted for cardiac regeneration in animal models, and the results are encouraging. However, there are still many hurdles to be overcome for the clinical application of embryonic stem cells. PMID- 16716099 TI - The emerging link between nutrition, inflammation and atherosclerosis. AB - A number of dietary components modulate the inflammatory response in humans, thereby affecting cardiovascular risk. As basic research provides a better understanding of the molecular mechanisms of vascular function regulation by nutrients, clinical investigation and outcome studies demonstrate the relevance of dietary factors to the prevention and treatment of vascular disease. Benefits of dietary interventions may be attributable to weight loss or to more specialized mechanisms in which inflammation is targeted directly. Available evidence indicates that dietary intervention should be an integral part of therapeutic approaches for treating conditions such as the metabolic syndrome and, ultimately, for the prevention of cardiovascular disease. PMID- 16716100 TI - Bilateral internal mammary artery grafting. AB - The superiority of the left internal mammary artery over the saphenous vein graft led many surgeons to adopt bilateral internal mammary artery (BIMA) as a good surgical option for further improving late outcome of patients undergoing myocardial revascularization. However, routine use of BIMA was limited by some potential drawbacks: the increase of deep sternal wound problems, especially in diabetic patients; the shortness of right internal mammary artery (RIMA), which limited its utilization as an in situ graft; the low patency rate if grafted to the right coronary artery; and the longer operative time. The skeletonization of the internal mammary artery along with a better glucose control in diabetic patients significantly reduced the incidence of deep sternal problems. Another problem to solve was finding the optimal target of the RIMA. The general consensus is that RIMA appears to be more efficient if grafted to the lateral wall. The Y or T configuration of double mammary arteries could be more helpful to reach the lateral target vessels. Finally, recent reports clearly demonstrate the superiority of BIMA over single internal mammary artery in terms of survival or quality of life. The latter finding has also been reported in diabetic patients. PMID- 16716101 TI - Role of heparin and low-molecular-weight heparins in the management of acute ischemic stroke. AB - The numerous large-scale randomized clinical trials performed during the last decade on either unfractionated heparin, or low molecular weight heparin have not been able to demonstrate undisputed benefits in patients with acute ischemic stroke, compared with no treatment or aspirin. However, a large number of these trials, including the International Stroke Trial and Chinese Acute Stroke Trial, exhibit severe methodological limitations and need to be interpreted with caution. Knowledge of thromboembolism pathophysiology and clinical experience leads to the theory that heparins will prevent red thrombus formation, propagation and embolism. Heparins effectively prevent venous thrombosis and pulmonary embolism. More trials are needed to test heparins in patients whose cardiocerebrovascular lesions are better defined by newer neuroimaging techniques. The efficacy of heparins has not been adequately tested in patients with defined stroke subtypes and occlusive vascular lesions. Heparins should not be indiscriminately given to all patients with acute ischemic stroke. High quality, randomized trials that adequately study heparin use in patients using modern technology for vascular lesions and stroke subtypes are lacking, and need to be performed. PMID- 16716103 TI - Antithrombotic therapy in heart failure. PMID- 16716102 TI - Role of beta-adrenergic receptor signaling and desensitization in heart failure: new concepts and prospects for treatment. AB - The use of beta-blockers to antagonize beta-adrenergic receptor signaling in the heart has become a standard method of treatment for heart failure, resulting in positive clinical outcomes alone and in conjunction with other modulators of cardiomyocyte contractility. However, an entire explanation for improved cardiac function in patients using beta-blockers is unknown, and in fact may be quite complicated, considering the numerous intracellular signaling pathways associated with beta-adrenergic receptors. Stimulation of beta-adrenergic receptors during both normal conditions and during heart failure activate several distinct signaling cascades, which influence cardiomyocyte contraction, hypertrophy and apoptosis. This review explores the signaling cascades induced by beta-adrenergic receptor activation in normal and desensitized states to provide new insight into the effective treatment of cardiac dysfunction. PMID- 16716105 TI - Lentivirus-mediated RNA interference therapy for human immunodeficiency virus type 1 infection. AB - RNA interference (RNAi) is a natural mechanism by which small interfering RNAs (siRNAs) operate to specifically and potently downregulate the expression of a target gene. This downregulation has been demonstrated by targeting siRNAs to the mRNA (posttranscriptional gene silencing) as well as to the gene promoter, regulating gene expression epigenetically by transcriptional gene silencing. These observations significantly broaden the role RNA plays in the cell and suggest that siRNAs could prove to be a potent future therapeutic for the treatment of diseases such as human immunodeficiency virus type 1 (HIV-1) infection. The specificity and simplicity of design and the ability to express siRNAs from mammalian promoters make the use of siRNAs to target and suppress virtually any gene or gene promoter of interest a soon-to-be-realized technology. However, the delivery and stable expression of siRNAs to target cells remain an enigma that could be surmounted, at least regarding the treatment of HIV-1 infection, by the application of lentiviral vectors to deliver and express anti HIV-1 siRNAs in target cells. This review focuses on the development, delivery, and potential therapeutic use of antiviral siRNAs in treating HIV-1. PMID- 16716106 TI - Correction of feline lipoprotein lipase deficiency with adeno-associated virus serotype 1-mediated gene transfer of the lipoprotein lipase S447X beneficial mutation. AB - Human lipoprotein lipase (hLPL) deficiency, for which there currently exists no adequate treatment, leads to excessive plasma triglycerides (TGs), recurrent abdominal pain, and life-threatening pancreatitis. We have shown that a single intramuscular administration of adeno-associated virus (AAV) serotype 1 vector, encoding the human LPL(S447X) variant, results in complete, long-term normalization of dyslipidemia in LPL(/) mice. As a prelude to gene therapy for human LPL deficiency, we tested the efficacy of AAV1-LPL(S447X) in LPL(/) cats, which demonstrate hypertriglyceridemia (plasma TGs, >10,000 mg/dl) and clinical symptoms similar to LPL deficiency in humans, including pancreatitis. Male LPL(/) cats were injected intramuscularly with saline or AAV1-LPL(S447X) (1 x 10(11)-1.7 x 10(12) genome copies [GC]/kg), combined with oral doses of cyclophosphamide (0 200 mg/m(2) per week) to inhibit an immune response against hLPL. Within 3-7 days after administration of >or=5 x 10(11) GC of AAV1-LPL(S447X) per kilogram, the visible plasma lipemia was completely resolved and plasma TG levels were reduced by >99% to normal levels (10-20 mg/dl); intermediate efficacy (95% reduction) was achieved with 1 x 10(11) GC/kg. Injection in two sites, greatly limiting the amount of transduced muscle, was sufficient to completely correct the dyslipidemia. By varying the dose per site, linear LPL expression was demonstrated over a wide range of local doses (4 x 10(10)-1 x 10(12) GC/site). However, efficacy was transient, because of an anti-hLPL immune response blunting LPL expression. The level and duration of efficacy were significantly improved with cyclophosphamide immunosuppression. We conclude that AAV1-mediated delivery of LPL(S447X) in muscle is an effective means to correct the hypertriglyceridemia associated with feline LPL deficiency. PMID- 16716107 TI - Efficacy of severe acute respiratory syndrome vaccine based on a nonhuman primate adenovirus in the presence of immunity against human adenovirus. AB - Replication-deficient human adenovirus type 5 (AdH5) vectors can induce strong transgene product-specific cellular and humoral responses. However, many adult humans have neutralizing antibodies (NAbs) against AdH5 as a result of natural infection with this virus. Therefore, a chimpanzee adenovirus C7 (AdC7) vector was developed to circumvent interference by preexisting immunity to AdH5. This study evaluated the impact of preexisting immunity to human adenovirus on the efficacy of adenovirus-based vaccines against the coronavirus that causes severe acute respiratory syndrome (SARS-CoV). Efficacy was assessed after intramuscular injection of the vector into mice and was measured as the frequency of SARS-CoV specific T cells and NAbs against SARS-CoV. Immunogenicity of the AdH5-based vaccine was significantly attenuated or completely abolished when the preexisting anti-AdH5 NAb titer was higher than 40. Because 27% of human serum samples from the United States tested so far have an anti-AdH5 NAb titer higher than 40, our results suggested that a significant percentage of humans with preexisting anti AdH5 immunity would not be candidates for vaccination with an AdH5-based genetic vaccine. In contrast, preexisting anti-AdH5 NAbs have a minimal effect on the potency of the AdC7-based genetic vaccine. Taken together, our studies warrant the further development of AdC7 as a vaccine carrier for human trials. PMID- 16716108 TI - Direct adenoviral transfer of bone morphogenetic protein-2 cDNA enhances fracture healing in osteoporotic sheep. AB - Osteoporosis, a major public health burden, is associated with increased fracture risk. Fracture healing in osteoporosis is delayed, with reduced callus formation and impaired biomechanical properties of newly formed bone leading to high risk of fixation failure. Adenoviral gene transfer of bone morphogenetic protein-2 (BMP-2) has been shown to enhance fracture healing. This study evaluated the ability of gene transfer to enhance bone healing in osteoporosis. An established sheep model of osteoporosis with well-characterized alterations in fracture healing was used. Osteotomies were created surgically in the tibias of adult female sheep and monitored for 8 weeks, using radiographic, biomechanical, and histological methods. For pilot experiments, primary ovine osteoblasts and mesenchymal stem cells were transduced with a recombinant adenovirus carrying BMP 2 cDNA (Ad.BMP-2). Large increases in alkaline phosphatase production and mineralization confirmed the ability of human BMP-2 to stimulate osteoblastic differentiation in sheep. In vivo bending stiffness measurements during fracture healing as well as ex vivo torsional stiffness measurements demonstrated stiffer callus tissue after treatment with Ad.BMP-2. The differences were found mainly in the early fracture-healing period. Computed tomography demonstrated that animals receiving the BMP-2 cDNA had larger cross-sectional callus area and higher callus density. Histological examination of the tibias confirmed enhanced callus formation. Direct, local adenoviral delivery of an osteogenic gene thus led to enhanced healing of fractures in an ovine model of osteoporosis. These promising data encourage the further development of genetic approaches to enhance bone healing in patients suffering osteoporosis-associated fractures. PMID- 16716109 TI - Coexpression of the uracil phosphoribosyltransferase gene with a chimeric human nerve growth factor receptor/cytosine deaminase fusion gene, using a single retroviral vector, augments cytotoxicity of transduced human T cells exposed to 5 fluorocytosine. AB - Donor T lymphocytes genetically engineered to express a "suicide gene" to facilitate negative selection represent a promising strategy for the management of graft-versus-host disease occurring after allogeneic hematopoietic cell transplantation (HCT). For this purpose, the herpes simplex virus thymidine kinase (HSV-tk) gene, although well studied, has limitations. Cytosine deaminase (CD), an alternative gene for negative selection, converts 5-fluorocytosine (5 FC) to the toxic metabolite 5-fluorouracil (5-FU). Sensitivity of cells to 5-FU can be further increased by expression of uracil phosphoribosyltransferase (UPRT), which catalyzes the conversion of 5-FU to 5-fluorouridine monophosphate. By using a chimeric gene (NG/CD) expressing the truncated human nerve growth factor receptor (NGFR) for positive selection fused to the Saccharomyces cerevisiae CD gene, we investigated strategies to achieve optimal T cell eradication by CD and UPRT expression, utilizing a single retroviral vector. Three vector strategies were compared on the basis of NGFR expression by flow cytometry, western analysis, and enzymatic activity. A construct (NG/CDiU) expressing UPRT and NG/CD, using a bicistronic message, provided the greatest UPRT activity and killing, reducing the lethal dose of 5-FC sufficient to eradicate 90% of cells from 38.7 microg/ml (300 microM) (NG/CD expression alone) to 0.13 microg/ml (1 microM). This approach provides an effective alternative to the HSV-tk system for eradication of donor T lymphocytes after allogeneic HCT. PMID- 16716111 TI - Reversal of renal fibrosis, inflammation, and glomerular hypertrophy by kallikrein gene delivery. AB - Evidence suggests that the progression of renal fibrosis is a reversible process. Because inflammation plays a crucial role in the development of renal injury, we examined the effect of kallikrein and activation of the kinin B2 receptor on the reversal of salt-induced inflammation and renal fibrosis in Dahl salt-sensitive (DSS) rats. Four weeks after high salt loading, when renal injury was apparent, adenovirus harboring the human tissue kallikrein gene was injected into DSS rats. To determine the role of the B2 receptor in mediating the actions of kallikrein, icatibant, a kinin B2 receptor antagonist, was infused with kallikrein gene delivery. Two weeks after adenovirus injection, salt-induced glomerular sclerosis, tubular protein cast formation, and monocyte/ macrophage accumulation in the kidney were notably reversed by kallikrein. Decreased intercellular adhesion molecule-1 expression paralleled this observation. Kallikrein gene delivery also dramatically reduced collagens I, III, and IV and reticulin deposition, accompanied by a decline in myofibroblast accumulation and transforming growth factor-beta(1) expression. Moreover, kallikrein reversed salt induced glomerular hypertrophy and inhibited the increase in levels of the cell cycle-inhibitory proteins p21 and p27. These protective actions of kallikrein were abolished by icatibant, indicating a B2 receptor-mediated event. In addition, kallikrein protected against salt-induced renal injury by diminishing urinary protein and blood urea nitrogen levels. Furthermore, kallikrein gene delivery restored nitric oxide production and suppressed NADH oxidase activity and superoxide generation. These results indicate that tissue kallikrein, through the kinin B2 receptor, reverses salt-induced inflammation, renal fibrosis, and glomerular hypertrophy via suppression of oxidative stress. PMID- 16716112 TI - CD46 represents a target for adenoviral gene therapy of malignant glioma. AB - Malignant gliomas remain refractory to adenovirus serotype 5 (Ad5) gene therapy because of the lack of the primary adenoviral receptor, the coxsackie-adenovirus receptor (CAR), on tumor cells. To bypass the dependence on CAR, we investigated the expression of adenovirus serotype 3 (Ad3) receptor, or CD46, on glioma cells. First, we analyzed the expression of CD46 by RT-PCR on primary and passaged glioma cells. We then performed immunofluorescence studies to examine protein expression of CAR and CD46 on the same tumor lines. Finally, we constructed a replication-defective Ad vector that binds to CD46 and contains a luciferase transgenic cassette in place of the deleted E1 region: Ad5/3 (containing tail/shaft domain of Ad5 and knob domain of Ad3). These vectors were analyzed in vitro and in vivo against malignant glioma and compared with wild-type Ad5 or control vector Ad3/5 (containing tail of Ad5, shaft of Ad3, and knob of Ad5). The chimeric vector Ad5/3 showed a significant increase in the transduction efficiency of glioma tumor cells. At the same time, blocking the CD46 receptor caused a 65% inhibition of adenoviral infection when using Ad5/3. Taken together, these results indicate that CD46 is overexpressed by malignant glioma. Retargeting to the Ad3 receptor enhances gene transfer and offers a novel target for gene therapy of malignant brain tumors. PMID- 16716110 TI - Quantification of high-capacity helper-dependent adenoviral vector genomes in vitro and in vivo, using quantitative TaqMan real-time polymerase chain reaction. AB - First-generation adenoviral (Ad) and high-capacity adenoviral (HC-Ad) vectors are efficient delivery vehicles for transferring therapeutic transgenes in vivo into tissues/organs. The initial successes reported with adenoviral vectors in preclinical trials have been limited by immune-related adverse side effects. This has been, in part, attributed to the use of poorly characterized preparations of adenoviral vectors and also to the untoward immune adverse side effects elicited when high doses of these vectors were used. HC-Ads have several advantages over Ads, including the lack of viral coding sequences, which after infection and uncoating, makes them invisible to the host's immune system. Another advantage is their large cloning capacity (up to approximately 35 kb). However, accurate characterization of HC-Ad vectors, and of contaminating replication-competent adenovirus (RCA) or helper virus, is necessary before these preparations can be used safely in clinical trials. Consequently, the development of accurate, simple, and reproducible methods to standardize and validate adenoviral preparations for the presence of contaminant genomes is required. By using a molecular method that allows accurate, reproducible, and simultaneous determination of HC-Ad, contaminating helper virus, and RCA genome copy numbers based on real-time quantitative PCR, we demonstrate accurate detection of these three genomic entities, within CsCl-purified vector stocks, total DNA isolated from cells transduced in vitro, and from brain tissue infected in vivo. This approach will allow accurate assessment of the levels and biodistribution of HC Ad and improve the safety and efficacy of clinical trials. PMID- 16716113 TI - Chimeric adeno-associated virus/antisense U1 small nuclear RNA effectively rescues dystrophin synthesis and muscle function by local treatment of mdx mice. AB - Duchenne muscular dystrophy (DMD) is a X-linked myopathy in which deletions and point mutations in the dystrophin gene abolish dystrophin expression. The defect can often be corrected at the posttranscriptional level by exon skipping. In an animal model of DMD, the mdx mouse, a point mutation in exon 23 of the dystrophin gene introduces a premature stop codon. Skipping of this exon reestablishes the open reading frame in the dystrophin mRNA. We have obtained persistent exon skipping in mdx mice by local muscle injection of AAV vectors expressing antisense sequences fused to either U1 or U7 small nuclear RNA (snRNA). In the transduced muscles, dystrophin expression, amelioration of muscle morphology, and significant force recovery were obtained. These data indicate that the expression of antisense snRNAs, combined with their efficient muscular delivery through AAV vectors, is a powerful strategy for the therapeutic treatment of DMD. Like U7 snRNA, spliceosomal U1 snRNA is also a suitable backbone for the expression of antisense molecules active in exon skipping. PMID- 16716116 TI - Regulation of human T-cell leukemia virus type 1 gene expression by Sp1 and Sp3 interaction with TRE-1 repeat III. AB - Transcription factors of the Sp family are known to play key roles in the regulation of both constitutive as well as cell type- and differentiation stage specific gene expression. Binding sites for factors of the Sp family (Sp1 and Sp3) have previously been identified within the U3 region of the human T-cell leukemia virus type 1 (HTLV-1) long terminal repeat (LTR). Although previous studies have demonstrated that Sp1 and Sp3 can interact with the Tax-responsive element 1 (TRE-1) repeat III, the sequences required for Sp1/Sp3 binding have not been mapped in detail. Herein, we demonstrate that the GC-rich regions flanking the viral cAMP-responsive element (CRE) within TRE-1 repeat III exhibit substantial affinity for both Sp1 and Sp3. We demonstrate that purified Sp1 competes with purified CREB for binding to TRE-1 repeat III due to the physical proximity of the Sp1/Sp3 and ATF/CREB binding sites, while purified Sp1 forms a multiprotein complex with purified CREB in the presence of Tax as demonstrated by electrophoretic mobility shift (EMS) analyses. Sp1 and Sp3 binding to the U3 region of the HTLV-1 LTR in the presence of Tax in vivo was confirmed by chromatin immunoprecipitation using HTLV-1-infected T cells (SLB-1 and C8166). Overexpression of Sp1 was modestly enhanced, while overexpression of Sp3 inhibited basal and Tax-mediated transactivation of the HTLV-1 LTR in U-937 cells (which express relatively low levels of endogenous Sp1 and Sp3). Furthermore, the modest upregulation of LTR activation caused by overexpression of Sp1 could be blocked by site-directed mutagenesis of the GC-rich Sp1/Sp3 binding sites within TRE-1 repeat III. These results suggest that both Sp1 and Sp3 transcription factor binding to TRE-1 repeat III participate in regulation of HTLV-1 viral gene expression. PMID- 16716115 TI - TLR9-independent activation of B lymphocytes by bacterial DNA. AB - The intracellular Toll-like receptor 9 (TLR9) is unique in its ability to recognize single-stranded DNA unmethylated at CpG motifs. Work from this laboratory showed that plasmid DNA is spontaneously internalized in B lymphocytes. This event is followed by the upregulation of costimulatory molecules and the acquisition of antigen presenting function by these cells. However, it is not known whether this phenomenon depends on TLR9. Because of the relevant role played by DNA-based drugs in immunotherapy and vaccination, and the central role of TLR9 signaling by CpG motifs, we decided to investigate whether signaling through TLR9 is a prerequisite for spontaneous transgenesis of lymphocytes. Here we found that transgene expression and upregulation of CD40 and CD86 costimulatory molecules was not inhibited by chloroquine treatment. Spontaneous transgenesis also occurred in B lymphocytes from TLR9-/- mice, and the injection of TLR9-/- transgenic B lymphocytes in C57Bl/6 mice induced both CD4 and CD8 T cell responses comparable to those induced by wild-type B lymphocytes. Collectively, these results suggest that plasmid DNA activates mammalian B lymphocytes through a TLR9 independent pathway. PMID- 16716117 TI - Antitumor therapeutic effects of e7 subunit and DNA vaccines in an animal cervical cancer model: antitumor efficacy of e7 therapeutic vaccines is dependent on tumor sizes, vaccine doses, and vaccine delivery routes. AB - We previously reported that E7 subunit and DNA vaccines are both capable of inducing antitumor protection through induction of antigen-specific CTL. In this study, we investigated their ability to control established tumors according to tumor size, vaccine doses, and vaccine delivery routes. Antitumor therapeutic efficacy of both vaccine types was dependent on tumor burden. However, E7 subunit vaccines induced a higher level of antitumor therapeutic activities at the tested dose compared to DNA vaccines. This was concomitant with induction of antibody, CTL, and IFN-gamma responses, as well as histologic changes (heavy infiltration of lymphocytes and presence of apoptotic bodies). In vaccine dose titration assays, 50 and 100 microg of DNA vaccines exhibited an equivalent antitumor efficacy to 0.5 and 1 microg of E7 subunit vaccines, respectively, i.e., a 100 fold difference in E7 dosage, suggesting the importance of vaccine doses for achieving antitumor immunity. Furthermore, tumors of a larger size were controlled by intratumoral injection with E7 subunit vaccines, underscoring the importance of vaccine delivery routes for antitumor therapeutic efficacy. Thus, these data suggest that antitumor therapeutic efficacy of E7 therapeutic vaccines is determined by vaccine doses, vaccine delivery routes, and tumor sizes, and that these vaccines could be another addition to conventional therapy modalities against cervical cancer. PMID- 16716118 TI - CYP17, SRD5A2, CYP1B1, and CYP2D6 gene polymorphisms with prostate cancer risk in North Indian population. AB - To investigate the involvement of the CYP17, SRD5A2, CYP1B1, and CYP2D6 variants with prostate cancer, a case-control study of 100 patients and an equal number of age-matched control men was conducted. There appears to be a nonsignificant increase with risk of prostate cancer for individuals carrying one copy of the CYP17 A2 allele (OR, 1.80; 95% CI, 0.99-3.29, P=0.05). The risk was increased in individuals having two A2 alleles (OR; 2.81, 95% CI, 1.06-7.40, P=0.03). Compared with men having the VV genotype of SRD5A2 gene, there was no significant association between the VL genotype and the risk of prostate cancer (OR; 0.54, 95% CI; 0.29-1.03, P=0.06). There was no difference in the occurrence of the genotype LL between controls and prostate cancer patients (OR; 0.90, 95% CI; 0.43 1.89, P=0.79). There was a nonsignificant increased risk of prostate cancer for individuals carrying the CYP1B1Leu/Val genotype (OR, 1.70, 95% CI, 0.91-3.17, P =0.09), which was increased in those having the Val/Val allele (OR, 3.38; 95% CI, 1.13-10.07, P=0.02). Relative to men homozygous for the wild-type allele in CYP2D6 gene, those heterozygous for the B allele had an odds ratio of 1.78 (95% CI, 0.76-4.17, P=0.18) for patients, and for homozygous individuals, it was 1.95 (0.55-6.93, P=0.30). These observations have suggested that the CYP17 A2/A2, CYP1B1 Val/Val, and CYP2D6 genotypes may be associated with an altered risk of prostate cancer, while the CYP2D6 and SRD5A2 V89L polymorphism have no association with its risk in the North Indian population. PMID- 16716119 TI - Common pathway of chromosome condensation in Mammalian cells. AB - Chromatin folding in the interphase nucleus is not known. We compared the pattern of chromatin condensation in Indian muntjac, Chinese hamster ovary, murine pre B, and K562 human erythroleukemia cells during the cell cycle. Fluorescent microscopy showed that chromosome condensation follows a general pathway. Synchronized cells were reversibly permeabilized and used to isolate interphase chromatin structures. Based on their structures two major categories of intermediates were distinguished: (1) decondensed chromatin and (2) condensed chromosomal forms. (1) Chromatin forms were found between the G1 and mid-S phase involving veil-like, supercoiled, fibrous, ribboned structures; (2) condensing chromosomal forms appeared in the late-S, G2, and M phase, including strings, chromatin bodies, elongated pre-chromosomes, pre-condensed chromosomes, and metaphase chromosomes. Results demonstrate that interphase chromosomes are clustered in domains; condensing interphase chromosomes are linearly arranged. Our results raise questions related to telomer sequences and to the chemical nature of chromosome connectivity. PMID- 16716120 TI - Novel immunoblotting monoclonal antibodies against human and rat CD36/fat used to identify an isoform of CD36 in rat muscle. AB - CD36, a surface membrane glycoprotein, functions as a class B scavenger receptor that binds to several distinct ligands. Included among these is oxidized low density lipoprotein (Ox-LDL), a major trigger of atherosclerotic lesions, and the levels of CD36 activity and Ox-LDL uptake may have an impact on coronary artery disease. In addition, recent studies in rodents have shown that CD36, also known as FAT, controls the levels of free fatty acids and triglycerides in plasma, and is an important regulator of the metabolic pathways involved in insulin resistance. Despite the importance of measuring CD36 expression in different tissues there is a paucity of good immunoblotting antibodies, particularly for rodent tissue. Here, using GST-fusion proteins incorporating the cysteine cluster encoded by exons VIII, IX, and X of the CD36 gene as immunogen, we have generated a panel of monoclonal antibodies that are excellent blotting reagents for human and rat CD36. With these reagents we were able to visualize an additional, faster migrating CD36 band in rat muscle, likely representing a minor splice variant of CD36 (CD36var.1) hitherto seen only in the human HEL cell line. PMID- 16716122 TI - Design of clinical trials of empiric antifungal therapy in patients with persistent febrile neutropenia: considerations and critiques. AB - Febrile neutropenic patients who receive antibiotics are at risk for fungal infections. This risk increases greatly with the length and severity of neutropenia. Because diagnostic tests for fungal infections lack sensitivity and specificity and because established fungal infections are associated with poor outcomes, empiric antifungal therapy is frequently given to patients with fever that persists despite antibacterial therapy. Early trials of empiric amphotericin B showed reductions in the number of invasive fungal infections and in related morbidity and mortality. However, as a result of infusion-related and renal adverse effects of amphotericin B, newer agents, such as lipid formulations of amphotericin B, extended-spectrum azoles, and echinocandins, have been developed. Although these alternatives have been associated with decreased toxicity, improved efficacy has not been clearly demonstrated. Although empiric antifungal therapy can prevent undetected breakthrough infections and morbidity associated with many fungal infections, its shortcomings include overtreatment, toxicity, and increased costs of unnecessary treatment. Recent studies have highlighted several questions in trial design and data interpretation. For example, what is the appropriate study design? Who should be enrolled in studies of empiric antifungal therapy? How should successful therapy be defined? These issues are reviewed to determine whether new antifungal agents should be evaluated for empiric use in patients with fever and neutropenia. PMID- 16716121 TI - Mitogen-activated protein kinase pathway was significantly activated in human bronchial epithelial cells by nicotine. AB - Nicotine is potentially associated with the onset of chronic obstructive pulmonary disease (COPD) and lung cancer. To gain insights into the molecular mechanism underlying such nicotine-induced conditions, microarray- bioinformatics analysis was carried out in the present study to explore the gene expression profiles in human bronchial epithelial cells (HBECs) treated with 5 microM nicotine for 4, 8, and 10 h. Of 1,800 assessed genes overall, 260 (14.4%) were upregulated and 17 (0.9%) down regulated significantly. Gene ontology analysis demonstrated that most of the differentially expressed genes belonged to the category of molecular function, especially to the subcategories of enzyme activity. The integration of obtained information with bioinformatics tools in DAVID and KEGG databases indicated that the greatest number of overexpressed genes was involved in mitogen-activated protein kinase (MAPK) pathway. Membrane array analysis subsequently suggested that both extracellular signal-regulated kinase (ERK) 1/2 and c-Jun-NH(2)-terminal kinase (JNK) signalings but not p38 MAPK signaling were activated in response to nicotine. Pretreatment of HBECs with specific inhibitors against ERK 1/2 and JNK but not p38 could significantly inhibit nicotine-induced interleukin- 8 production. These results suggest that MAPK pathway may mediate the effect of nicotine through ERK 1/2 and JNK but not p38 in HBECs treated with nicotine. PMID- 16716123 TI - Treatment options for invasive fungal infections. AB - Clinicians face an increasing occurrence of invasive fungal infections. These are due not only to traditional yeast and mould species but also to rare pathogens that can be difficult to treat. The introduction of new agents has expanded the options for treating common and rare mycotic infections with antifungal efficacy at least equal, and safety far superior, to that of a once-limited choice of therapies. Patients with invasive mycoses frequently have concomitant disorders and require multidrug regimens. Clinicians must be aware of the potential for interactions among agents available for treating invasive mycoses in patients with serious underlying conditions. PMID- 16716124 TI - Decision making in antifungal monotherapy versus combination therapy. AB - The availability of new antifungal agents with novel mechanisms of action and improved tolerability has widened the possibilities for combination therapy for difficult-to-treat opportunistic mycoses. However, empiricism largely governs this therapy, especially in patients with invasive mould infections for whom there is a dire need to improve outcomes. Because of difficulties associated with the design and conduct of clinical trials of combination therapy for opportunistic mycoses, most studies are still performed in the laboratory or in animal models. Methods to assess combined antifungal effects in vitro and in animals are poorly standardized, and little evidence suggests that the data generated from these studies are relevant to treating patients. Even without solid evidence to support routine combination therapy, certain principles can guide its use in select patients. PMID- 16716125 TI - Candida resistance and its clinical relevance. AB - Since the 1970s, the death rate from Candida infection has risen in conjunction with increasing numbers of patients at risk for serious fungal infections, such as those immunocompromised because of tissue or organ transplantation, chemotherapy, acquired immunodeficiency syndrome, or advanced age. Candidal infections are not only prevalent but also associated with considerable mortality and morbidity. In 2005, the overall mortality rate was 44% within 30 days of the first blood culture positive for any Candida species. Given the substantial morbidity and mortality, clinicians must include fungal infection in the differential diagnosis for at-risk patients, and they must quickly select appropriate antifungal therapy for those with identified infection. However, clinicians cannot use the minimum inhibitory concentration to select antifungal therapy in the same way they use it to treat bacterial infections. The relationship between in vitro susceptibility and clinical effect is not as direct with antifungals as it is with antibiotics. As long as Candida species continue to be the major causes of fungal infections, improving outcomes remains an important therapeutic goal. A key element is preventing and managing drug resistance. Further study of treatment duration, dosage, intermittent-versus continuous administration schedules, and other treatment options are needed to determine their effect on resistance. Therapy that combines agents with complementary mechanisms of action may increase potency and broaden the spectrum of antifungal efficacy while decreasing the number of resistant organisms. PMID- 16716127 TI - Clinical pharmacy services, pharmacy staffing, and adverse drug reactions in United States hospitals. AB - Adverse drug reactions (ADRs) were examined in 1,960,059 hospitalized Medicare patients in 584 United States hospitals in 1998. A database was constructed from the MedPAR database and the National Clinical Pharmacy Services survey. The 584 hospitals were selected because they provided specific information on 14 clinical pharmacy services and on pharmacy staffing; they also had functional ADR reporting systems. The study population consisted of 35,193 Medicare patients who experienced an ADR (rate of 1.8%). Of the 14 clinical pharmacy services, 12 were associated with reduced ADR rates. The most significant reductions occurred in hospitals offering pharmacist-provided admission drug histories (odds ratio [OR] 1.864, 95% confidence interval [CI] 1.765-1.968), drug protocol management (OR 1.365, 95% CI 1.335-1.395), and ADR management (OR 1.360, 95% CI 1.328-1.392). Multivariate analysis, performed to further evaluate these findings, showed that nine variables were associated with ADR rate: pharmacist-provided in-service education (slope -0.469, p=0.018), drug information (slope -0.488, p=0.005), ADR management (slope -0.424, p=0.021), drug protocol management (slope -0.732, p=0.002), participation on the total parenteral nutrition team (slope 0.384, p=0.04), participation on the cardiopulmonary resuscitation team (slope -0.506, p=0.008), medical round participation (slope -0.422, p=0.037), admission drug histories (slope -0.712, p=0.008), and increased clinical pharmacist staffing (slope -4.345, p=0.009). As clinical pharmacist staffing increased from the 20th to the 100th percentile (from 0.93+/-0.77/100 to 5.16+/-4.11/100 occupied beds), ADRs decreased by 47.88%. In hospitals without pharmacist-provided ADR management, the following increases were noted: mean number of ADRs/100 admissions by 34.90% (OR 1.360, 95% CI 1.328-1.392), length of stay 13.64% (Mann Whitney U test [U]=11047367, p=0.017), death rate 53.64% (OR 1.574, 95% CI 1.423 1.731), total Medicare charges 6.88% (U=111298871, p=0.018), and drug charges 8.16% (U=108979074, p<0.001). Patients in hospitals without pharmacist-provided ADR management had an excess of 4266 ADRs, 443 deaths, 85,554 patient-days, $11,745,342 in total Medicare charges, and $1,857,744 in drug charges. The implications of these findings are significant for our health care system, especially considering that the study population represented 15.55% of 12,261,737 Medicare patients and 5.71% of the 34,345,436 patients admitted to all U.S. hospitals. PMID- 16716128 TI - Atypical antipsychotics and pituitary tumors: a pharmacovigilance study. AB - STUDY OBJECTIVE: To analyze the disproportionality of reporting of hyperprolactinemia, galactorrhea, and pituitary tumors with seven widely used antipsychotic drugs. DESIGN: Retrospective pharmacovigilance study. DATA SOURCE: United States Food and Drug Administration's Adverse Event Reporting System (AERS) database. INTERVENTION: We initially identified higher-than-expected postmarketing reports of pituitary tumors associated with risperidone, a potent dopamine D2-receptor antagonist antipsychotic, by analyzing reporting patterns of these tumors in the AERS database. To further examine this association, we analyzed disproportionate reporting patterns of pituitary tumor reports for seven antipsychotics with different affinities for blocking D2 receptors: aripiprazole, clozapine, olanzapine, quetiapine, risperidone, ziprasidone, and haloperidol. MEASUREMENTS AND MAIN RESULTS: To conduct both of these analyses, we used the Multi-item Gamma Poisson Shrinker (MGPS) data mining algorithm applied to the AERS database. The MGPS uses a Bayesian model to calculate adjusted observed:expected ratios of drug-adverse event associations (Empiric Bayes Geometric Mean [EBGM] values) in huge drug safety databases. The higher the adjusted reporting ratio, or EBGM value, the greater the strength of the association between a drug and an adverse event. Risperidone had the highest adjusted reporting ratios for hyperprolactinemia (EBGM 34.9, 90% confidence interval [CI] 32.8-37.1]), galactorrhea (EBGM 19.9, 90% CI 18.6-21.4), and pituitary tumor (EBGM 18.7, 90% CI 14.9-23.3) among the seven antipsychotics, and one of the highest scores for all drugs in the AERS database. Some tumors were associated with visual field defects, hemorrhage, convulsions, surgery, and severe (>10-fold) prolactin elevations. The EBGM values for risperidone for these adverse events were higher in women, but high EBGM values for these events were also seen in men and children. Moreover, the rank order of the EBGM values for pituitary tumors corresponded to the affinities of these seven drugs for D2 receptors. CONCLUSION: Treatment with potent D2-receptor antagonists, such as risperidone, may be associated with pituitary tumors. These findings are consistent with animal (mice) studies and raise the need for clinical awareness and longitudinal studies. PMID- 16716129 TI - Scientific collaboration results in higher citation rates of published articles. AB - STUDY OBJECTIVE: The primary objective was to analyze the relationship between the citation rate of an article and the extent of collaboration. The secondary objective was to analyze the relationship between the number of authors/article and the number of institutions/article for the period of study. METHODS: We counted the number of original research articles published in six leading journals--Cell, Science, Nature, New England Journal of Medicine, The Lancet, and Journal of the American Medical Association--for the years 1975, 1985, and 1995. For each article, we determined the number of authors and the number of separate institutions. We also determined the number of times each article that was published in 1995 was cited in future scientific articles from the Science Citation Index database. RESULTS: Science, Cell, Nature, New England Journal of Medicine, The Lancet, and Journal of the American Medical Association had 2014, 868, 3856, 643, 785, and 465 total articles published/3-year study period, respectively. There was a median of 2, 2, 2, 3, 3, and 3 institutions/article, respectively. All of the final models had a significant linear author component for which all of the parameter estimates were positive, yet variable. Thus, the number of times an article was cited correlated significantly with the number of authors and the number of institutions. CONCLUSION: A correlation exists between the number of authors and the number of times an article is cited in other articles. Investigators who are open to collaborations and those who seem to adequately manage those collaborations produce a superior product that results in a higher impact. PMID- 16716130 TI - Potentially inappropriate drug use and health-related quality of life in the elderly. AB - STUDY OBJECTIVE: To investigate the relationship between two widely used, generic health-related quality of life (HRQOL) measures, Short Form-12 (SF-12) and EuroQol's EQ-5D, and potentially inappropriate drug use in an elderly cohort. DESIGN: Longitudinal retrospective cohort study. DATA SOURCE: Medical Expenditure Panel Survey, panel 5. PARTICIPANTS: Respondents aged 65 years or older. MEASUREMENTS AND MAIN RESULTS: Participants with potentially inappropriate drug use were identified by using National Drug Codes based on the Beers updated criteria. The dependent variable, HRQOL, was measured by using self-reported scores on the SF-12 (physical component summary, mental component summary, and global rating of health) and the EQ-5D (index and visual analog scale). The HRQOL data were regressed onto scores for potentially inappropriate drug, age, sex, number of prescriptions, race, comorbidity, and previous HRQOL. Regression analysis revealed statistically significant models for all five HRQOL equations (adjusted R2=26.50-53.83%, p<0.0001). However, potentially inappropriate drug use was not a significant predictor of HRQOL in any of the models tested. Previous HRQOL, as measured by using the SF-12 (global, physical component summary, and mental component summary) and the EQ-5D (index and visual analog scale), was a significant predictor of HRQOL; number of prescriptions was also a significant predictor of HRQOL, as measured by using the SF-12 (global and physical component summary) and the EQ-5D (index and visual analog scale). CONCLUSION: The results supported others showing that a significant proportion of the elderly receive care that is potentially harmful and not supported by evidence-based practice. Use of a disease-specific HRQOL scale may be more responsive to measuring the impact of potentially inappropriate drugs on patients' HRQOL. PMID- 16716131 TI - Association of refill adherence and health care use among adults with hypertension in an urban health care system. AB - STUDY OBJECTIVES: To determine the rates of undersupply, appropriate supply, and oversupply of antihypertensive drugs, as measured by refill adherence, among patients with complicated and uncomplicated hypertension (i.e., patients who have and have not, respectively, experienced hypertension-related target organ damage), and to examine the association of refill adherence with hospitalization and health care costs among these patients. DESIGN: Retrospective analysis of electronic medical records. SETTING: An urban, public health care system. PATIENTS: A total of 15,206 patients aged 18 years or older whose electronic medical records indicated a clinical diagnosis of hypertension based on the International Classification of Diseases, Ninth Revision, Clinical Modification codes, and who had received at least one prescription of an antihypertensive drug from 1995-2001. MEASUREMENTS AND MAIN RESULTS: We used multivariable analyses to investigate the association of refill adherence with hospitalization and costs. On average, 53% of patients had appropriate supplies (80-120% of supplies needed), 7% had undersupplies, and 40% had oversupplies of drug annually. For patients with complicated hypertension, an undersupply of drug was associated with a 15% greater probability (p=0.009) and an oversupply with a 16% greater probability (p<0.0001) of hospitalization. Among those with uncomplicated hypertension, oversupply was associated with an 11% greater probability (p=0.0002) of hospitalization; undersupply was not associated with greater probability of hospitalization. Total health care costs were lower for patients with undersupplies and higher for those with oversupplies of drug. CONCLUSION: Among adults in an urban health care system with complicated hypertension, both undersupply and oversupply of drug were associated with increased hospitalization rates. Monitoring refill adherence of patients, particularly those with low income, minority status, and complicated hypertension, may be useful for targeting patients with undersupplies of drug to encourage refill adherence and identifying patients with oversupplies, who are at high risk of hospitalization. PMID- 16716132 TI - Population pharmacokinetics of cyclosporine in chinese cardiac transplant recipients. AB - STUDY OBJECTIVE: To characterize the population pharmacokinetics of cyclosporine in Chinese patients undergoing cardiac transplantation and to identify the demographic and clinical covariates affecting cyclosporine clearance. DESIGN: Population pharmacokinetic analysis using data from a retrospective chart review. SETTING: Specialty hospital in Hong Kong for treatment of cardiac and pulmonary diseases. PATIENTS: Thirty-eight Chinese adult patients (mean age 46 yrs) who had undergone routine cyclosporine therapeutic drug monitoring after cardiac transplantation between January 1, 1991, and December 31, 2003. MEASUREMENTS AND MAIN RESULTS: Data regarding dosing, demographics, clinical laboratory values, and concurrent drugs were collected retrospectively. Data were included if patients had blood cyclosporine concentrations determined for at least 12 weeks after transplantation; an average of 18 blood samples/patient were collected. Population modeling was performed using a one-compartment linear model with first order absorption and elimination. Various demographic and clinical covariates were tested for their significant effects on the apparent oral clearance (Cl/F) of cyclosporine. The stability of the final population model was evaluated by using the bootstrap resampling method. Statistically significant associations were observed between Cl/F and each of the following covariates: body weight (BW), use of diltiazem (DIL), and hematocrit value (HCT). The final model was Cl/F=5.00*(1-DIL)+365/HCT+(0.144*BW). The interindividual variabilities of Cl/F and apparent volume of distribution were 14.5% and 40.2%, respectively. The mean parameter estimates obtained from bootstrap analyses were highly consistent with those obtained with the original data set. CONCLUSION: The estimated Cl/F values of cyclosporine in our Chinese cardiac transplant recipients appeared to be similar to those reported for Caucasian cardiac transplant recipients. Thus, our data provide support that a cyclosporine dosage regimen similar to that in Caucasian patients may be needed in Chinese cardiac transplant recipients. However, further studies are required to determine the optimum cyclosporine dosage regimen in the Chinese population. PMID- 16716133 TI - Addition of dexmedetomidine to standard sedation regimens after cardiac surgery: an outcomes analysis. AB - STUDY OBJECTIVE: To characterize inpatient use of intravenous sedatives in the real-world setting, and to evaluate clinical and economic outcomes when dexmedetomidine was used with midazolam and propofol for select cardiovascular procedures. DESIGN: 12-month retrospective analysis. DATA SOURCE: An administrative claims database of operational data from a nationally representative sample of 250 medical and surgical hospitals. PATIENTS: Patients who received midazolam plus propofol (9996 patients) or dexmedetomidine, midazolam, plus propofol (356 patients) after cardiac valve or vessel surgery. MEASUREMENTS AND MAIN RESULTS: The source of patient demographics (e.g., age, sex, Charlson Comorbidity Index) and outcomes (e.g., charges, length of stay, mortality rate) was the hospital billing claim form. Patients in the dexmedetomidine-midazolam-propofol cohort tended to be younger and male and to have fewer comorbidities than those midazolam-propofol cohort. The primary outcomes for the three-drug cohort showed significant reductions in total charges/patient (approximately $18,000, p<0.05), total hospital length of stay (0.6 days, p<0.0001), days in the intensive care unit or cardiac care unit (3.87 days, p<0.0001), and mortality (2%, p=0.0142). Although pharmacy charges were higher (approximately $4000/patient), lower charges for the intensive care or cardiac care unit, operating room, room and board, and respiratory services were observed in the dexmedetomidinemidazolam-propofol cohort compared with the two drug cohort. Also, mechanical ventilation was shorter by approximately 0.5 day in the three-drug cohort (p<0.01). CONCLUSION: These initial findings of a real world assessment of dexmedetomidine use with other agents suggest favorable clinical and economic outcomes. Further research through randomized clinical trials of dexmedetomidine is warranted to better understand its optimum patient population, dosage, and the causality of the results, and to confirm the potential clinical and economic benefits observed in our patients. PMID- 16716134 TI - Single-dose rasburicase 6 mg in the management of tumor lysis syndrome in adults. AB - Rasburicase is currently approved at a dosage of 0.15-0.2 mg/kg once/day for 5 days in pediatric patients with cancer to lower plasma uric acid concentrations and manage tumor lysis syndrome (TLS). Information on rasburicase dosing in adults is limited, with some data on using rasburicase as a single dose instead of multiple daily doses. Therefore, we evaluated the efficacy of a single dose of rasburicase for preventing or managing TLS in adults. We collected retrospective data for 11 adults with hematologic malignancies who received a single 6-mg dose of rasburicase. All patients received intravenous hydration with urinary alkalinization and allopurinol; however, due to adverse reactions, two patients received short courses of allopurinol. Only patients at high risk for TLS (e.g., large tumor burden, increasing uric acid concentration) or those with TLS received rasburicase. The single dose of rasburicase 6 mg resulted in a median 0.0773-mg/kg dose (range 0.0232-0.1361 mg/kg). The single 6-mg dose rapidly lowered uric acid concentrations in 10 of the 11 patients. The median uric acid concentration of 11.7 mg/dl (range 7.4-17.4 mg/dl) declined to 2.0 mg/dl (range 0.5-15.4 mg/dl) within a day after rasburicase administration (p=0.022). In these 10 patients, uric acid concentrations remained low despite subsequent chemotherapy, and none required additional rasburicase doses. The only patient who did not respond to the single 6-mg rasburicase dose was a morbidly obese man (259 kg, body mass index 87 kg/m2) who subsequently responded to an additional dose of rasburicase 12 mg. These results warrant further investigation of a single 6-mg dose of rasburicase in adults with TLS or at high-risk for developing TLS. PMID- 16716135 TI - Pharmacokinetics of intravenous immunoglobulin: a systematic review. AB - BACKGROUND: Intravenous immunoglobulin (IGIV) is used in the treatment of a wide variety of immune disorders. To our knowledge, no comprehensive or systematic review on the pharmacokinetics of IGIV has been published despite the availability of many published individual studies. OBJECTIVE: To systematically review published studies of the pharmacokinetics of IGIV. METHODOLOGY: We conducted a search of PubMed/MEDLINE from January 1966-September 2005 and EMBASE from January 1980-September 2005 for English-language articles on the pharmacokinetics of IGIV. This search was supplemented by a bibliographic review of all relevant articles. RESULTS: Data elements extracted from these articles included study design, number of study subjects, indication for IGIV therapy, IGIV treatment regimen (formulation, dosage, and duration), pharmacokinetic parameters (clearance, volume of distribution, elimination rate constant, and half-life), analytic methodology, pharmacokinetic model, and blood sampling times. The United States Preventive Services Task Force rating scale was used to categorize the 50 pertinent citations identified in our literature search. According to the rating scale, 12 studies were level I (prospective, randomized, controlled studies), 3 were level II-1 (prospective, nonrandomized, controlled studies), 30 were level II-2 (prospective, nonrandomized, uncontrolled [cohort] studies), and 5 were level III (case reports or descriptive studies). CONCLUSION: The pharmacokinetics of IGIV shows considerable intra- and interpopulation variability among patients with normal immunoglobulin levels, patients with primary immunodeficiency diseases, bone marrow transplant recipients, patients with immune deficiency due to chronic lymphocytic leukemia or multiple myeloma, very low birth weight neonates, neonates with suspected sepsis, high-risk infants in the neonatal intensive care unit, high-risk infants with cardiopulmonary disease, children with cryptogenic West or Lennox-Gastaut syndrome, women and infants with fetal alloimmune thrombocytopenia, and women with recurrent spontaneous abortions. Despite the large number of studies characterizing the pharmacokinetics of IGIV, major literature gaps include lack of information on IGIV clearance or area under the curve parameters and target serum immunoglobulin G concentrations. Further study is needed to rigorously characterize the pharmacokinetic properties of IGIV in a range of patient populations. PMID- 16716136 TI - Evolving role of vasopressin in the treatment of cardiac arrest. AB - Sudden cardiac arrest is a major public heath problem, affecting more than 450,000 individuals annually. Response time and the initiation of cardiopulmonary resuscitation (CPR) remain the most important factors determining successful revival. During resuscitation, sympathomimetics are given to enhance cerebral and coronary perfusion pressures in an attempt to achieve restoration of spontaneous circulation. Epinephrine has been the preferred vasopressor since the inception of advanced cardiac life support, although the lack of definitive evidence regarding its effectiveness has created much controversy surrounding its use, including the optimum dosage. Vasopressin is an alternative vasopressor that, when given at high doses, causes vasoconstriction by directly stimulating smooth muscle V1 receptors. The 2000 American Heart Association (AHA) guidelines commented that vasopressin is a reasonable first-line vasopressor in patients with ventricular fibrillation or pulseless ventricular tachycardia. Since release of those guidelines, additional human studies support an expanded role for vasopressin, whereas other studies cast doubt regarding its efficacy compared with epinephrine. The AHA recently released revised guidelines for CPR and emergency cardiovascular care. The consensus was that vasopressors should remain a part of pulseless sudden cardiac arrest management, with epinephrine 1 mg every 3-5 minutes being the recommended adrenergic of choice. In these revised guidelines, the role of vasopressin expanded beyond previous recommendations, despite the recommendation being downgraded to class indeterminate. The guidelines comment that one dose of vasopressin 40 U may replace the first or second dose of epinephrine in all pulseless sudden cardiac arrest scenarios, including asystole and pulseless electrical activity. A consistent theme with all vasopressors in sudden cardiac arrest is that additional studies are necessary to clearly document greater efficacy compared with no treatment. Further evaluation is warranted to better assess the role of vasopressin in asystolic sudden cardiac arrest, as well as its use with epinephrine, and to determine its optimal timing of administration and potential synergistic effects. PMID- 16716137 TI - Update on apomorphine for the rapid treatment of hypomobility ("off") episodes in Parkinson's disease. AB - As Parkinson's disease progresses, fluctuations between akinesia, or hypomobility ("off" times), and mobility ("on" times) increase in frequency despite optimized pharmacotherapy. Motor fluctuations include predictable shortening of therapeutic effects, nocturnal or early morning akinesia, random hypomobility, and delayed mobility (variable responses to individual doses of drugs). Current oral antiparkinson drugs are inadequate for rapid and consistent relief of symptoms during hypomobility. Apomorphine, an injectable dopamine agonist recently introduced in the United States, is indicated for the management of hypomobility associated with advanced Parkinson's disease. Subcutaneous apomorphine is effective for rapid and consistent rescue from hypomobility, with a magnitude of motor improvement similar to that of levodopa. The effect begins within 20 minutes after dosing and lasts approximately 100 minutes. Therapeutic rescue doses are 2-6 mg, and patients typically require approximately three rescue doses/day. Apomorphine is associated with a clinically significant potential to cause nausea and orthostatic hypotension. These potential effects can be managed with antiemetic prophylaxis and appropriate determination of the therapeutic rescue dose. PMID- 16716138 TI - Docetaxel-associated epiphora. AB - Docetaxel is a semisynthetic taxane indicated for the treatment of advanced breast, prostate, and non-small cell lung cancers; it is also used for the treatment of various other solid tumors. The standard intermittent dosage of docetaxel is 60-100 mg/m2 every 3 weeks. At this dose and schedule, myelosuppression is common and neutropenia is usually the dose-limiting toxicity. Weekly administration of docetaxel 20-42 mg/m2 is being tested in the treatment of advanced solid tumors in order to improve patient tolerance by reducing the interval dose and to maintain therapeutic efficacy by increasing overall dose intensity. Asthenia and peripheral neuropathy can limit continued administration of weekly docetaxel. Epiphora (excess tearing due to narrowing or blockage of the lacrimal outflow passages) is associated with repeated weekly administration of docetaxel. This adverse effect can interfere with activities of daily life and negatively affect quality of life. Epiphora may be an underreported adverse effect of treatment because of underrecognition by clinicians and patient embarrassment with respect to seemingly uncontrolled tearing. The use of weekly docetaxel administration is expanding; therefore, patients should be educated to recognize and report signs and symptoms of epiphora. It is important for clinicians participating in the care of patients undergoing treatment with docetaxel to monitor for excess tearing and signs of eye irritation to ensure timely management of treatment-related epiphora. PMID- 16716139 TI - Leflunomide and warfarin interaction: case report and review of the literature. AB - A 61-year-old Caucasian woman receiving long-term anticoagulation with warfarin for recurrent thromboembolism and atrial fibrillation was found to have an elevated international normalized ratio (INR) after she started leflunomide therapy for rheumatoid arthritis. Her INR had been stable for 4 months before this event. The patient required an overall decrease of 22% in her weekly warfarin dose to maintain a therapeutic INR within the goal range of 2.0-3.0 after adding leflunomide therapy. A comprehensive PubMed/MEDLINE search was conducted to identify literature addressing the potential interaction between warfarin and leflunomide. Evidence describing the interaction and its potential mechanism was limited to one published case report and to in vitro data, respectively. Our case report provides additional support that such an interaction exists and that it was at least partly responsible for the subsequent increase in the patient's INR. Therefore, continued evaluation and documentation of this potential drug interaction is imperative. To reduce the risk of adverse effects related to excessive anticoagulation with the start of leflunomide in patients taking warfarin, clinicians should increase their frequency of INR monitoring and adjust the warfarin dosage accordingly to maintain therapeutic anticoagulation. PMID- 16716140 TI - Telithromycin-induced digoxin toxicity and electrocardiographic changes. AB - A 58-year-old woman who had been taking digoxin 0.25 mg/day for more than 35 years for heart palpitations after mitral valve repair was prescribed a 5-day course of telithromycin for acute bronchitis. On the sixth day of therapy, she came to the emergency department complaining of general malaise and having experienced three episodes of syncope over the previous 2 days. Laboratory analysis revealed elevated digoxin plasma levels, and electrocardiography showed several nonspecific repolarization anomalies. Telithromycin is known to increase digoxin plasma levels; however, the clinical significance of this interaction is not known. To our knowledge, this is the first report of elevated plasma digoxin levels associated with signs and symptoms of toxicity. This drug interaction determined as probable according to the Naranjo adverse drug reaction probability scale-may be mediated by P-glycoprotein. By inhibiting the transport of digoxin by P-glycoprotein, telithromycin may have decreased digoxin elimination in the intestinal lumen and its renal tubular excretion, resulting in elevated plasma levels and drug toxicity. Clinicians should be aware of possible digoxin toxicity after concomitant administration with telithromycin, especially in patients who are at risk, such as those with electrolyte abnormalities and decreased renal function. PMID- 16716141 TI - Progressive esophagitis caused by Candida albicans with reduced susceptibility to caspofungin. AB - Candida esophagitis, a defining illness of acquired immunodeficiency syndrome (AIDS), requires systemic antifungal therapy. Candida albicans can become resistant to commonly administered azole antifungal agents. An attractive alternative is caspofungin, an echinocandin antifungal that has generally displayed predictable activity against C. albicans. We report the case of a 29 year-old woman with AIDS who developed recurrent esophagitis caused by a strain of C. albicans that showed reduced susceptibility to caspofungin (elevated minimum inhibitory concentration of 8 mg/L). Analysis of the strain revealed that it contained a serine-to-proline substitution at position 645 in the FKS1 gene. Clinicians who prescribe caspofungin to treat esophagitis caused by C. albicans should recognize the potential risk, albeit slight, for acquired resistance to caspofungin and possibly other echinocandin antifungal agents in the face of persistent disease. In patients who are refractory or unresponsive to caspofungin therapy, susceptibility testing and/or alternative therapy should be considered. PMID- 16716142 TI - Increased thyroid-stimulating hormone levels associated with concomitant administration of levothyroxine and raloxifene. AB - A 47-year-old Caucasian woman had a 3.5-year history of primary hypothyroidism treated with levothyroxine. Her levothyroxine dosage of 0.05 mg/day had been stable for the past 15 months. She was then prescribed raloxifene for prevention of osteoporosis secondary to early menopause. During the next 30 months, her levothyroxine dosage had to be gradually increased. The patient had been taking levothyroxine and raloxifene at the same time each day on an empty stomach. During the months of her levothyroxine dosage changes, however, she separated administration of levothyroxine and raloxifene by 12 hours; the patient then became hyperthyroid. Eventually, her levothyroxine needs decreased, and she returned to the same levothyroxine dosage she had taken before separating administration of the two drugs. These findings suggest that raloxifene decreased the absorption of levothyroxine when the two agents were coadministered. Assessment of causality using the Naranjo adverse drug reaction probability scale resulted in a possible association for this adverse event. Another published case report provides findings similar to our patient's experience. The possibility of a malabsorption interaction between levothyroxine and raloxifene is significant, as hypothyroidism is common among postmenopausal women-the same population that is the target of osteoporosis therapy with agents such as raloxifene. The mechanism by which raloxifene decreases levothyroxine absorption is unknown. Further investigation of this potential interaction is warranted. Until then, clinicians should be alert to the potential for an interaction between raloxifene and levothyroxine. PMID- 16716143 TI - Reduction of high-risk polypharmacy drug combinations in patients in a managed care setting. PMID- 16716144 TI - WITHDRAWN:Chimeric hypoxanthine guanine phosphoribosyltransferases: Effect of segment swapping on xanthine specificity and tetramer stability. AB - The paper entitled "Chimeric hypoxanthine guanine phosphoribosyltransferases: Effect of segment on xanthine specificity and tetramer stability", which was published online on 22 May 2006, was withdrawn at the author's request. PMID- 16716145 TI - Integrin signalling regulates the nuclear localization and function of the lysophosphatidic acid receptor-1 (LPA1) in mammalian cells. AB - We show that LPA1 (lysophosphatidic acid receptor-1) is constitutively localized in the nucleus of mammalian cells. LPA1 also traffics from cell membranes to the nucleus in response to LPA (lysophosphatidic acid). Several lines of evidence suggest an important role for cell-matrix interaction in regulating the constitutive nuclear localization of LPA1. First, the RGDS peptide, which blocks cell matrix-induced integrin clustering and cytoskeletal rearrangement, reduced the number of cells containing LPA1 in the nucleus. Secondly, a higher proportion of cells contained nuclear LPA1 when adhesion on fibronectin-coated glass was compared with adherence to polylysine-coated glass. Thirdly, pre-treatment of cells with the Rho kinase inhibitor (Y27632) or the myosin light chain kinase inhibitor (ML9) reduced the number of cells containing nuclear LPA1. The addition of LPA and/or Ki16425 (which binds to LPA1) to isolated nuclei containing LPA1 induced the phosphorylation of several proteins with molecular masses of 34, 32, 14 and 11 kDa. These findings demonstrate that trafficking of LPA1 to the nucleus is influenced by cell-matrix interactions and that nuclear LPA1 may be involved in regulating intranuclear protein phosphorylation and signalling. PMID- 16716146 TI - HIV-1 integrase is capable of targeting DNA to the nucleus via an importin alpha/beta-dependent mechanism. AB - In addition to its well-documented role in integration of the viral genome, the HIV-1 enzyme IN (integrase) is thought to be involved in the preceding step of importing the viral cDNA into the nucleus. The ability of HIV to transport its cDNA through an intact nuclear envelope allows HIV-1 to infect non-dividing cells, which is thought to be crucial for the persistent nature of HIV/AIDS. Despite this, the mechanism utilized by HIV-1 to import its cDNA into the nucleus, and the viral proteins involved, remains ill-defined. In the present study we utilize in vitro techniques to assess the nuclear import properties of the IN protein, and show that IN interacts with members of the Imp (Importin) family of nuclear transport proteins with high affinity and exhibits rapid nuclear accumulation within an in vitro assay, indicating that IN possesses potent nucleophilic potential. IN nuclear import appears to be dependent on the Imp alpha/beta heterodimer and Ran GTP (Ran in its GTP-bound state), but does not require ATP. Importantly, we show that IN is capable of binding DNA and facilitating its import into the nucleus of semi-intact cells via a process that involves basic residues within amino acids 186-188 of IN. These results confirm IN as an efficient mediator of DNA nuclear import in vitro and imply the potential for IN to fulfil such a role in vivo. These results may not only aid in highlighting potential therapeutic targets for impeding the progression of HIV/AIDS, but may also be relevant for non-viral gene delivery. PMID- 16716147 TI - Functional expression of a biologically active fragment of soluble gp130 as an ELP-fusion protein in transgenic plants: purification via inverse transition cycling. AB - In murine models of Crohn's disease, rheumatoid arthritis and colon cancer, IL-6 (interleukin-6) signalling via the sIL-6R (soluble IL-6 receptor; termed IL-6 trans-signalling) has been shown to promote the pathology associated with these conditions. These detrimental activities can, however, be selectively blocked by soluble forms of the gp130 (glycoprotein 130) receptor. Although sgp130 (soluble gp130) therefore represents a viable therapeutic modality for the treatment of these conditions, the mass manufacture of such biologics is often expensive. The advent of molecular farming has, however, provided an extremely cost-effective strategy for the engineering of recombinant proteins. Here, we describe the expression and production of a biologically active sgp130 variant that is expressed in transgenic tobacco plants as an ELP (elastin-like peptide)-fusion protein (mini-gp130-ELP). Mini-gp130-ELP consists of the first three domains of gp130 (Ig-like domain and cytokine binding module) fused to 100 repeats of ELP. Expression of mini-gp130-ELP did not affect the growth rate or morphology of the transgenic plants, and purification was achieved using inverse transition cycling. This approach led to an overall yield of 141 microg of purified protein per g of fresh leaf weight. The purified mini-gp130-ELP specifically inhibited sIL-6R-mediated trans-signalling as measured by binding to the IL-6-sIL-6R complex and through its ability to block sIL-6R-mediated activation of STAT3 (signal transducer and activator of transcription 3) phosphorylation and proliferation in human hepatoma cells and murine pre-B-cells. Consequently, the present study validates the potential application of molecular farming in transgenic tobacco plants as a strategy for the expression and purification of therapeutically advantageous biologics such as sgp130. PMID- 16716149 TI - Identification and functional characterization of hCLS1, a human cardiolipin synthase localized in mitochondria. AB - In eukaryotic cells, CLS (cardiolipin synthase) is involved in the final step of cardiolipin synthesis by catalysing the transfer of a phosphatidyl residue from CDP-DAG (diacylglycerol) to PG (phosphatidylglycerol). Despite an important role of cardiolipin in regulating mitochondrial function, a gene encoding the mammalian CLS has not been identified so far. We report in the present study the identification and characterization of a human cDNA encoding the first mammalian CLS [hCLS1 (human CLS1)]. The predicted hCLS1 peptide sequence shares significant homology with the yeast and plant CLS proteins. The recombinant hCLS1 enzyme expressed in COS-7 cells catalysed efficiently the synthesis of cardiolipin in vitro using CDP-DAG and PG as substrates. Furthermore, overexpression of hCLS1 cDNA in COS-7 cells resulted in a significant increase in cardiolipin synthesis in intact COS-7 cells without any significant effects on the activity of the endogenous phosphatidylglycerophosphate synthase of the transfected COS-7 cells. Immunohistochemical analysis demonstrated that the recombinant hCLS1 protein was localized to the mitochondria when transiently expressed in COS-7 cells, which was further corroborated by results from subcellular fractionation analyses of the recombinant hCLS1 protein. Northern-blot analysis showed that the hCLS1 gene was predominantly expressed in tissues that require high levels of mitochondrial activities for energy metabolism, with the highest expression in skeletal and cardiac muscles. High levels of hCLS1 expression were also detected in liver, pancreas, kidney and small intestine, implying a functional role of hCLS1 in these tissues. PMID- 16716150 TI - Rituximab in refractory autoimmune bullous diseases. AB - Treatment of autoimmune blistering diseases consists of systemic glucocorticosteroids usually in combination with additional immunosuppressants such as azathioprine and mycophenolate mofetil or immunomodulators such as dapsone, antibiotics, intravenous immunoglobulins, and immunoadsorption. In some patients, these treatment regimens are not sufficient to control disease activity and/or lead to intolerable adverse events. Rituximab, originally developed for the treatment of non-Hodgkin's lymphoma, is an anti-CD20 humanized monoclonal antibody leading to transitory B-cell depletion. For this indication, rituximab is widely employed, and severe side-effects rarely observed. Subsequently, the B cell-depleting effect of rituximab has been exploited successfully in various autoimmune disorders, including autoimmune blistering diseases. Here, we review the effect of rituximab in such diseases. To date, application of rituximab has been reported in 26 treatment-resistant patients with the vulgaris, foliaceus, and paraneoplastic variants of pemphigus as well as in bullous pemphigoid and epidermolysis bullosa acquisita. All but a single patient showed clinical improvement with reduction of lesion formation. In about a third, a clinical remission requiring further immunsuppressive medication was achieved, and in about a quarter, complete remission was induced. In addition, the mode of action and adverse events of rituximab as well as adjuvant immunosuppressive treatments, and the effect on levels of circulating autoantibodies in these patients are discussed. PMID- 16716148 TI - Inhibition of MEK/ERK1/2 signalling alters endothelial nitric oxide synthase activity in an agonist-dependent manner. AB - eNOS (endothelial nitric oxide synthase) activity is post-translationally regulated in a complex fashion by acylation, protein-protein interactions, intracellular trafficking and phosphorylation, among others. Signalling pathways that regulate eNOS activity include phosphoinositide 3-kinase/Akt, cyclic nucleotide-dependent kinases [PKA (protein kinase A) and PKG], PKC, as well as ERKs (extracellular-signal-regulated kinases). The role of ERKs in eNOS activation remains controversial. In the present study, we have examined the role of ERK1/2 in eNOS activation in HUVEC-CS [transformed HUVEC (human umbilical-vein endothelial cells)] as well as a widely used model for eNOS study, transiently transfected COS-7 cells. U0126 pretreatment of HUVEC-CS potentiated ATP stimulated eNOS activity, independent of changes in intracellular Ca2+ concentration ([Ca2+]i). In COS-7 cells transiently expressing ovine eNOS, U0126 potentiated A23187-stimulated eNOS activity, but inhibited ATP-stimulated activity. Compensatory changes in phosphorylation of five key eNOS residues did not account for changes in A23187-stimulated activity. However, in the case of ATP, altered phosphorylation and changes in [Ca2+]i may partially contribute to U0126 inhibition of activity. Finally, seven eNOS alanine mutants of putative ERK1/2 targets were generated and the effects of U0126 pretreatment on eNOS activity were gauged with A23187 and ATP treatment. T97A-eNOS was the only construct significantly different from wild-type after U0126 pretreatment and ATP stimulation of eNOS activation. In the present study, eNOS activity was either potentiated or inhibited in COS-7 cells, suggesting agonist dependence for MEK/ERK1/2 signalling [where MEK is MAPK (mitogen-activated protein kinase)/ERK kinase] to eNOS and a complex mechanism including [Ca2+]i, phosphorylation and, possibly, intracellular trafficking. PMID- 16716152 TI - Oesophageal involvement during attacks in pemphigus vulgaris patients. AB - BACKGROUND: Pemphigus vulgaris (PV) is a rare autoimmune bullous skin disorder characterized by frequent involvement of the mucous membranes, usually beginning at the mouth. AIMS: To investigate the oesophageal involvement in patients with PV and to explore the primary relationship of the disease with symptoms such as dysphagia, odynophagia and retrosternal burning. METHODS: Oesophageal involvement was investigated by upper gastrointestinal endoscopy and biopsy during the early phase of the attacks in 26 patients with PV (12 men, 14 women, age range 24-63 years). RESULTS: Histopathological examination and direct immunofluorescence of the oesophageal biopsy specimens revealed pemphigus involvement in 12 of 26 patients (46.15%). CONCLUSION: The oesophagus is an important predilection zone for PV, thus care must be taken to detect these lesions at an early stage. PMID- 16716151 TI - The impact of immunosuppression on erythema migrans. A retrospective study of clinical presentation, response to treatment and production of Borrelia antibodies in 33 patients. AB - BACKGROUND: Little is known about the potential influence of immunosuppression on erythema migrans, the hallmark of early Lyme borreliosis. METHODS: We performed a retrospective study to assess the impact of immunosuppression on erythema migrans in 33 patients with a malignant or autoimmune disease, chronic infection, or immunosuppressive therapy for organ transplantation. Only patients with active disease status and/or current immunosuppressive therapy were included. Pre treatment clinical parameters, such as presentation of the skin lesion and presence of extracutaneous signs and symptoms, the disease course during a median follow-up of 9 months after therapy and serum anti-Borrelia burgdorferi antibodies before therapy and by the end of follow-up in the 33 immunosuppressed patients were statistically compared with 75 otherwise healthy patients with erythema migrans. The 75 control patients were matched for sex, age and antibiotic therapy. RESULTS: With the exception of the site of erythema migrans lesions, which were found more often on the trunk than on the legs in the immunosuppressed patients (vice versa in immunocompetent patients), we found no significant differences for all investigated parameters between the two groups. CONCLUSIONS: It appears that immunosuppression does not influence clinical presentation, response to therapy, or production of anti-B. burgdorferi antibodies of patients with erythema migrans. It is thus not necessary to treat immunosuppressed patients with erythema migrans differently from immunocompetent patients. PMID- 16716153 TI - Methotrexate and ciclosporin combination for the treatment of severe psoriasis. AB - BACKGROUND: Psoriasis is a chronic skin disease that often requires long-term therapy to control the symptoms. Combination therapies for severe psoriasis have advantages for disease control and are thought to reduce long-term side-effects. OBJECTIVE: To assess the efficacy and side-effects of methotrexate plus ciclosporin used in combination for the treatment of severe psoriasis. METHODS: In this prospective study, 20 patients were treated with the combination of methotrexate and ciclosporin. Methotrexate was given intramuscularly as a single weekly dose of 10 mg and ciclosporin at a dose of 3.5 mg/kg/day in two equally divided doses. Clinical response was assessed according to clinical outcome and the Psoriasis Area and Severity Index which were evaluated at the beginning of therapy (PASI1), after cessation of one agent (PASI2), and at the end of therapy (PASI3). RESULTS: All the patients had previously received one or more systemic treatment. There were 10 women and 10 men (mean age 44 years). The median (minimum to maximum) duration of methotrexate and ciclosporin treatments were 12.5 (4-55) and 14.0 (4-80) weeks, respectively. Median duration of combination therapy was 9.5 weeks (range 4-50). The median of previously used and end-of study cumulative doses of methotrexate were 181.8 mg (range 0-785) and 330.8 mg (range 50-845), respectively. The median PASI scores were decreased by 77.4% (range 51.2-90.2) and 75.9% (range 10.1-100) at PASI2 and PASI3, respectively. CONCLUSION: Patients with severe psoriasis had clinically significant improvement after the initiation of combination therapy. Healing rate was decreased upon cessation of one of the medications. Short-term side-effects were minor, transient and manageable. Long-term follow-up of patients treated with this combination is needed. PMID- 16716154 TI - Successful use of a thermoplastic dressing in two cases of the trigeminal trophic syndrome. AB - Treatment of trigeminal trophic syndrome is challenging and often unsatisfactory, particularly in patients with cognitive impairment. We report the novel use of a thermoplastic dressing in two patients with trigeminal trophic syndrome. Use of the dressing resulted in successful healing of ulceration in both patients, which has been maintained in the short term, representing a simple and economical therapeutic option in the management of this difficult condition. PMID- 16716155 TI - Calibre persistent labial artery: clinical features and noninvasive radiological diagnosis. AB - Calibre persistent labial artery (CPLA) is defined as a primary arterial branch that penetrates into the submucosal tissue without division or decrease in diameter. It usually presents as an asymptomatic papule on the lower lip and can be easily misdiagnosed as a varix, haemangioma, venous lake, mucocele or fibroma. When it is ulcerated, squamous cell carcinoma is the most usual differential diagnosis. The most frequently used method to confirm the diagnosis of CPLA has been excisional biopsy, which carries the risk of profuse bleeding. Angiography, another invasive method, has also been used. Here, we report a case of a 20-year old woman with a 5-year history of multiple CPLA lesions involving both upper and lower lips. In this case, the diagnosis was made clinically and confirmed by Doppler ultrasonography, which is a noninvasive and simple diagnostic tool. PMID- 16716156 TI - Familial pachydermoperiostosis in association with protein-losing enteropathy. AB - In this report we describe a rare association of pachydermoperiostosis with protein-losing enteropathy (PLE) in a family of three brothers. The first brother had the complete form of pachydermoperiostosis along with PLE. The second brother had the 'forme fruste' of pachydermoperiostosis, with minimal skin changes, bony abnormalities and PLE, which was due to intestinal lymphangiectasia. The third brother had an incomplete form of pachydermoperiostosis without evidence of PLE. To our knowledge, the association of pachydermoperiostosis with PLE due to intestinal lymphangiectasia has not been reported previously. PMID- 16716157 TI - Possible isotretinoin-induced keloids in a patient with Behcet's disease. AB - Isotretinoin-induced keloid formation has occasionally been reported in patients who have undergone dermabrasion or laser treatment. This report describes a man with Behcet's disease (BD) who was prescribed isotretinoin for nodulocystic acne. After approximately 8 weeks of this treatment, the patient developed extensive keloids. There are no previous reports of possible isotretinoin-induced keloid formation in the setting of BD. This article discusses possible connections between BD, keloid formation and isotretinoin, which may be manifested in this case. PMID- 16716158 TI - Large plaque-like glomuvenous malformation (glomangioma) simulating venous malformation. AB - Glomuvenous malformations and venous malformations are vascular lesions that can be distinguished on the basis of clinical and pathological features. A vascular lesion of the skin and superficial and deep soft tissues of a lower limb in a 5 year-old child is described. The clinical and radiological features, including skeletal muscle involvement, were typical of venous malformation, whereas the histopathological features were those of a glomuvenous malformation. The clinical and histopathological features are briefly discussed. PMID- 16716159 TI - Squamous cell carcinoma developing in two Chinese patients with chronic discoid lupus erythematosus: the need for continued surveillance. AB - Squamous cell carcinoma (SCC) is a rare late sequel of chronic discoid lupus erythematosus (CDLE). We report two cases of SCC developing in Chinese patients with CDLE. The first patient had prior biopsies from the same site that showed viral warts and the second patient had multiple histologically confirmed viral warts around the vicinity of the previously excised SCC. In this paper, we emphasize the need to be suspicious of warty lesions on skin afflicted by CDLE, and repeat biopsies should be performed if there is failure to respond to conventional therapy. PMID- 16716160 TI - Cutaneous abscess by Trichosporon asahii developing on a steroid injection site in a healthy adult. AB - We report a rare case of cutaneous abscess by Trichosporon asahii in an immunocompetent adult. A 31-year-old Korean woman presented to our hospital with a cutaneous abscess. She had received an intralesional steroid injection 4 months earlier on the site of a hypertrophic scar. Direct sequencing of the intergenic spacer regions of the rRNA genes identified T. asahii. The decreased local immunity after the steroid injection might have triggered the infection by T. asahii. A cutaneous abscess formation by T. asahii in an immunocompetent patient is an unusual cutaneous finding that to our knowledge has not been reported previously. The local immune reaction of the skin is important for the prevention of Trichosporon infection. PMID- 16716161 TI - The role of mitochondria in ageing and carcinogenesis. AB - Mitochondria can perform multiple cellular functions including energy production, cell proliferation and apoptosis. These organelles contain their own genetic material, mitochondrial DNA (mtDNA), which is maternally inherited. Although much smaller than the nuclear genome, mtDNA is equally important, as it has been hypothesized to play a crucial role in ageing and carcinogenesis. This is partly due to the fact that mitochondria represent the major site for the generation of cellular oxidative stress and play a key role in mediating programmed cell death (apoptosis). Damage to mtDNA is therefore an important contributor to human ageing, cancer and neurodegenerative diseases. The most relevant footprints of mtDNA damage are point mutations of single bases, or deletions of the 16.5-kb mitochondrial genome. This review will focus on the key roles of mitochondrial function and mtDNA in oxidative stress production and as a mediator of apoptosis, and on the use of mtDNA as a biomarker of sun exposure. This will be related to the contribution of mitochondria and mtDNA in the ageing process and cancer, with a specific focus on human skin. In conclusion, it is likely that the interplay between nuclear and mitochondrial genes may hold the final understanding of the mitochondrial role in these disease processes. PMID- 16716162 TI - A novel single-nucleotide polymorphism of the Fcgamma receptor IIIa gene is associated with genetic susceptibility to systemic lupus erythematosus in Chinese populations: a family-based association study. AB - BACKGROUND: Systemic lupus erytematosus (SLE) is characterized by the presence of various autoantibodies and the deposition of immune complexes which are cleared by Fcgamma receptors. OBJECTIVES: Family-based association analysis was performed to investigate whether the FCGR3A-72S/R and FCGR3A-270T/R polymorphisms are risk factors for SLE in a Chinese population. METHODS: In total, 119 patients with SLE from 95 nuclear families, aged 14-78 years, who met the American College of Rheumatology 1997 criteria were recruited, as were 316 family members of these patients. We studied two single-nucleotide polymorphisms (SNPs) encoding nonsynonymous substitution in the FCGR3A gene with respect to genetic susceptibility to SLE in a collection of 435 subjects from 95 nuclear families. We performed the genotyping using PCR restriction fragment length polymorphism. RESULTS: Our results showed that FCGR3A-72R/S have an excess of transmission of the R allele from heterozygous parents to affected offspring (transmission disequilibrium test chi2 = 9.30, P = 0.0032). Univariate (single-marker) family based association tests demonstrated that a variant allele at SNP rs403016 of the FCGR3A gene was significantly associated with genetic susceptibility to SLE (exon 3, Z = 2.5444, P = 0.01097) in an additive model. The R and S allele frequencies were 39.4% and 60.6%, respectively. The frequencies of FCGR3A 72R/R, R/S and SS genotypes were 9.1%, 60.6% and 30.3%, respectively. However, the FCGR3A-270T/S SNP was not found in this Chinese population. CONCLUSION: This study suggests a linkage disequilibrium of the FCGR3A-72R/S SNP with SLE, and supports the notion that a novel polymorphism of the FCGR3A-72R/S SNP is associated with genetic susceptibility to SLE in Chinese populations. PMID- 16716163 TI - Genetic heterogeneity in acrokeratosis verruciformis of Hopf. AB - BACKGROUND: Acrokeratosis verruciformis of Hopf (AKV) is a rare genodermatosis characterized by multiple flat-topped, flesh-coloured papules on the dorsa of hands and feet, and punctuate keratoses on the palms and soles. A mutation in the ATP2A2 gene has been shown to be associated with AKV and with Darier's disease (DD). OBJECTIVES: To explore the molecular aetiology of AKV and DD. METHODS: We investigated the clinical and histological information in two families and a sporadic case with AKV and one family and a sporadic case with DD in China. Mutation analysis of ATP2A2 was performed by PCR and direct sequencing, and genotyping and linkage analysis performed using six polymorphic microsatellite markers spanning the locus at 12q23-12q24 containing ATP2A2. RESULTS: Mutational analysis showed no mutation in ATP2A2 among the AKV patients, but we found two novel mutations (p.C318F and p.M719fs) in the DD patients. The genotyping and linkage analysis results revealed no linkage evidence of the locus at 12q23-12q24 in a large AKV family. CONCLUSIONS: Our findings provide evidence for the genetic heterogeneity of AKV and demonstrate that mutations in genes other than ATP2A2 are responsible for AKV in a proportion of the Chinese population. PMID- 16716164 TI - Content of 5-alpha-reductase (type 1 and type 2) mRNA in dermal papillae from the lower abdominal region in women with hirsutism. AB - BACKGROUND: Androgens influence the growth of terminal hair. The dermal papilla contains androgen receptors and the enzymes 5-alpha-reductase types 1 and 2. Both of these enzymes convert testosterone to the more active androgen, 5-alpha dihydrotestosterone. The male distribution pattern of terminal hair in females is termed hirsutism. It is most common among women with hyperandrogenism; however, it may also affect patients with normal androgen levels (idiopathic hirsutism). OBJECTIVES: The aim of this study was to assess the expression of 5-alpha reductase types 1 and 2 mRNA in dermal papillae from the lower abdominal skin in women with hirsutism. METHODS: The study included 42 subjects, 24 with a diagnosis of polycystic ovary syndrome (PCOS) and 18 with idiopathic hirsutism (IH). In all patients, free serum testosterone was measured. RESULTS: The mean +/ SD concentration of free serum testosterone was 7.2 +/-5.3 pmol/L in the total group of patients, 10.8 +/- 4.0 pmol/L in patients with PCOS, and 2.5 +/- 1.7 pmol/L in patients with IH. Quantitative analysis was then performed for the mRNA of 5-alpha-reductase types 1 and 2, both of which were found within the dermal papillae from the lower abdominal skin region. The number of mRNA copies/microg of total RNA for 5-alpha-reductase type 1 was statistically significantly higher than that for type 2 in both groups of examined patients. We also demonstrated a positive correlation between the number of mRNA copies/microg of total RNA for 5 alpha-reductase types 1 and 2 and the concentration of free serum testosterone in women with PCOS and IH. Considering all patients together, we found a positive correlation between the number of mRNA copies/microg of total RNA for 5-alpha reductase type 2 and the concentration of free serum testosterone. There was also a tendency towards a positive correlation between the number of mRNA copies/microg of total RNA for 5-alpha-reductase type 1 and the concentration of free serum testosterone. CONCLUSION: The results of our study suggest that testosterone increases expression of 5-alpha-reductase types 1 and 2 in dermal papillae from the lower abdominal region in patients with hirsutism. PMID- 16716165 TI - Endothelin-1 is significantly elevated in plasma of patients with vitiligo treated with psoralen plus ultraviolet A. AB - BACKGROUND: Recent evidence suggests that systemic psoralen plus ultraviolet A (PUVA) therapy may have a stimulatory effect on melanocytes, not only locally but also systemically. Aim. We aimed to assess endothelin-1 (ET-1), a potent melanocyte mitogen, in plasma of PUVA-treated paients with vitiligo. METHODS: ET 1 was sequentially assessed (using ELISA) in patients with nonsegmental vitiligo treated with PUVA (n = 20), at 8, 16 and 24 h following the PUVA session. Evaluations took place at 0, 1 and 3 months of therapy. Patients with psoriasis (n = 15) treated identically and healthy subjects not receiving any therapy (n = 15) served as controls. Vitiligo Area Scoring Index (VASI) and Psoriasis Area Severity Index (PASI) scores were simultaneously evaluated. RESULTS: ET-1 was significantly lower in vitiligo than in psoriasis at month 0 (8.2 +/- 3.6 vs. 13.7 +/- 5.4 pg/mL; P = 0.03) and it was significantly higher in both than in healthy controls at all time points of the PUVA sessions (P < 0.001). In vitiligo, it significantly increased at month 3 at 8 (8.2 +/- 3.6 vs. 10.8 +/- 2.7 pg/mL; P = 0.02) and 16 h (8.2 +/- 3.6 vs. 11.5 +/- 3.9 pg/mL; P < 0.01), whereas in psoriasis, it significantly decreased at month 3 at 8 (13.7 +/- 5.4 vs. 3.5 +/- 0.4 pg/mL; P < 0.01) and 16 h (13.7 +/- 5.4 vs. 6.3 +/- 4 pg/mL; P = 0.01). In contrast to psoriasis, sequential values of vitiligo revealed insignificant variance (P > 0.05). VASI score significantly decreased at month 3 (19 +/- 9.6 vs. 11.9 +/- 7.3; P < 0.01), whereas PASI score significantly decreased at months 1 (38.2 +/- 16.1 vs. 13.8 +/- 3; P < 0.05) and 3 (38.2 +/- 16.1 vs. 7 +/- 2.6; P = 0.03). There was a significant indirect correlation of ET 1 with VASI score (P < 0.01) and a significant direct correlation with PASI score (P < 0.01). CONCLUSION: Systemic PUVA therapy in vitiligo may have a generalized mitogenic effect on melanocytes through the release of ET-1 into the circulation. PMID- 16716166 TI - Type I interferon-associated skin recruitment of CXCR3+ lymphocytes in dermatomyositis. AB - BACKGROUND: Dermatomyositis (DM) is an autoimmune disease of unknown origin affecting skin and muscles. Infiltrating autoreactive T lymphocytes are thought to play an important pathogenetic role, but it is unclear which mechanisms are involved in the recruitment of these cells. Recent studies provided evidence that a type I interferon (IFN)-driven immune response, including the recruitment of T cells via IP10/CXCR3 interactions, might be important for the generation of skin lesions of cutaneous lupus erythematosus (CLE), an autoimmune disease that shares some clinical and histopathological features with DM. We hypothesized that a similar mechanism might also be involved in the pathogenesis of DM skin lesions. METHODS: Skin biopsies of 23 donors (11 DM, 5 healthy controls, 7 CLE controls) were analysed by immunohistochemistry using monoclonal antibodies against CD3, CD4, CD8, CD20, CD68, CD123, the chemokine receptor CXCR3 and its ligand IP10/CXCL10, and the myxovirus-resistance protein A (MxA)-protein, which is a specific marker for type I IFNs. RESULTS: We detected strong expression of the MxA protein in all DM skin biopsies, indicating involvement of type I IFNs. Expression of MxA was closely associated with expression of the interferon inducible protein IP10/CXCL10 and the recruitment of CXCR3+ lymphocytes. Plasmacytoid dendritic cells appear to be an important source of type I IFNs in DM. DISCUSSION: Our results support the hypothesis that lesional type I IFN signalling, induction of IP10 expression, and recruitment of potentially autoreactive T cells via IP10/CXCR3 interaction are involved in the pathogenesis of DM skin lesions. PMID- 16716167 TI - Serum substance P and calcitonin gene-related peptide levels in Behcet's disease and their association with disease activity. AB - BACKGROUND: Substance P (SP) and calcitonin gene-related peptide (CGRP) are neuropeptides that have a role in several cutaneous diseases and inflammations. Aim. To evaluate SP, CGRP and serum interleukin (IL)-8 levels in Behcet's disease (BD) and to explore the relationship of these peptides with BD activity. METHODS: The study group comprised 30 patients with BD, and 30 healthy individuals acted as controls. Serum levels of SP, CGRP and IL-8 were determined by micro-ELISA test during the active and inactive disease periods of patients with BD. These data were compared with each other and controls. Active and inactive periods of BD were established. RESULTS: The mean +/- SD serum CGRP (ng/ml) and IL-8 levels (pg/ml) in inactive BD (5.87 +/- 2.49 and 0.62 +/- 0.24, respectively) were significantly higher than the control group (4.74 +/- 1.17 and 0.46 +/- 0.11) (P < 0.05 for both). The difference between serum CGRP and IL-8 levels in active BD were also significantly higher than in inactive BD (P < 0.05 for both). Serum SP values (ng/ml) in active BD (18.27 +/- 5.38) were significantly higher than in inactive BD (15.26 +/- 5.74) and controls (12.6 +/- 4.45) (P < 0.05 for all), whereas the difference between the serum SP values in inactive BD and the control group was not statistically significant (P > 0.05). CONCLUSION: Serum SP and CGRP may have a role in the pathogenesis of BD. In addition, serum IL-8, SP and CGRP levels can be used as laboratory parameters indicating activity in BD. PMID- 16716169 TI - Oral anticoagulants may prevent NSAID-induced urticaria. PMID- 16716168 TI - Severe cholinergic urticaria successfully treated with scopolamine butylbromide in addition to antihistamines. PMID- 16716170 TI - Phakomatosis pigmentovascularis type II b associated with Sturge-Weber syndrome. PMID- 16716172 TI - Bullous cellulitis and myonecrosis secondary to Escherichia coli in a patient with cirrhosis. PMID- 16716171 TI - Digitate dermatosis responding to bicalutamide therapy. PMID- 16716173 TI - Acute myeloid leukaemia presenting with eczema. PMID- 16716174 TI - An unusual case of an acquired acral partial lipodystrophy (Barraquer-Simons syndrome) in a patient with extrinsic allergic alveolitis. PMID- 16716175 TI - The eclipse naevus and cockade naevus: are they two of a kind? PMID- 16716176 TI - Onychotillomania, major depressive disorder and suicide. PMID- 16716177 TI - Vitamin B12 and folate deficiency anaemia associated with isotretinoin treatment for acne. PMID- 16716178 TI - A case of follicular keratosis with trichostasis on an amputated limb. PMID- 16716179 TI - Pustular psoriasis over keloids: is it a Koebner phenomenon? PMID- 16716180 TI - An old classic. PMID- 16716181 TI - Recurrent blistering of the scalp with scarring. PMID- 16716182 TI - An unusual nodular lesion. PMID- 16716183 TI - Crusted lesions in a baby. PMID- 16716184 TI - Papular tuberculides post-BCG vaccination: case report and review of the literature in Japan. PMID- 16716185 TI - Giant bullous cutaneous larva migrans. PMID- 16716186 TI - A brother and sister with Werner's syndrome demonstrating extensive tendon calcification and sacroiliitis. PMID- 16716188 TI - Cell cycle execution point analysis of ORC function and characterization of the checkpoint response to ORC inactivation in Saccharomyces cerevisiae. AB - Chromosomal replication initiates through the assembly of a prereplicative complex (pre-RC) at individual replication origins in the G1-phase, followed by activation of these complexes in the S-phase. In Saccharomyces cerevisiae, the origin recognition complex (ORC) binds replication origins throughout the cell cycle and participates in pre-RC assembly. Whether the ORC plays an additional role subsequent to pre-RC assembly in replication initiation or any other essential cell cycle process is not clear. To study the function of the ORC during defined cell cycle periods, we performed cell cycle execution point analyses with strains containing a conditional mutation in the ORC1, ORC2 or ORC5 subunit of ORC. We found that the ORC is essential for replication initiation, but is dispensable for replication elongation or later cell cycle events. Defective initiation in ORC mutant cells results in incomplete replication and mitotic arrest enforced by the DNA damage and spindle assembly checkpoint pathways. The involvement of the spindle assembly checkpoint implies a defect in kinetochore-spindle attachment or sister chromatid cohesion due to incomplete replication and/or DNA damage. Remarkably, under semipermissive conditions for ORC1 function, the spindle checkpoint alone suffices to block proliferation, suggesting this checkpoint is highly sensitive to replication initiation defects. We discuss the potential significance of these overlapping checkpoints and the impact of our findings on previously postulated role(s) of ORCs in other cell cycle functions. PMID- 16716189 TI - Predictive base substitution rules that determine the binding and transcriptional specificity of Maf recognition elements. AB - Small Maf transcription factors possess a basic region-leucine zipper motif through which they form homodimers or heterodimers with CNC and Bach proteins. Different combinations of small Maf and CNC/Bach protein dimers bind to cis acting DNA elements, collectively referred to as Maf-recognition elements (MAREs), to either activate or repress transcription. As MAREs defined by function are often divergent from the consensus sequence, we speculated that sequence variations in the MAREs form the basis for selective Maf:Maf or Maf:CNC dimer binding. To test this hypothesis, we analyzed the binding of Maf-containing dimers to variant sequences of the MARE using bacterially expressed MafG and Nrf2 proteins and a surface plasmon resonance-microarray imaging technique. We found that base substitutions in the MAREs actually determined their binding preference for different dimers. In fact, we were able to categorize MAREs into five groups: MafG homodimer-orientd MAREs (Groups I and II), ambivalent MAREs (Group III), MafG:Nrf2 heterodimer-orientd MAREs (Group IV), and silent MAREs (Group V). This study thus manifests that a clear set of rules pertaining to the cis-acting element determine whether a given MARE preferentially associates with MafG homodimer or with MafG:Nrf2 heterodimer. PMID- 16716190 TI - Clathrin anchors deubiquitinating enzymes, AMSH and AMSH-like protein, on early endosomes. AB - Endosomal sorting of ubiquitinated membrane proteins for trafficking to lysosomes is executed by a complex of two ubiquitin-binding proteins, Hrs and STAM, that localizes on a microdomain of early endosomes with a flat clathrin coat. AMSH is a deubiquitinating enzyme that interacts with STAM and is implicated in the down regulation of epidermal growth factor receptor. AMSH has a close homolog, AMSH like protein (AMSH-LP). Here we show that AMSH-LP is also a deubiquitinating enzyme that acts on early endosomes. We further show that AMSH and AMSH-LP bind to the terminal domain of clathrin heavy chain via a novel clathrin-binding site conserved between these proteins. Exogenously expressed AMSH and AMSH-LP co localized with clathrin on early endosomes. However, deletion of the clathrin binding site from the proteins, as well as RNA interference-mediated depletion of clathrin heavy chain, resulted in a failure of AMSH and AMSH-LP to localize on endosomes. In contrast, a mutant of AMSH that lacks the ability to bind STAM localized normally on endosomes. We suggest that AMSH and AMSH-LP are anchored on the early endosomal membrane via interaction with the clathrin coat. PMID- 16716191 TI - Role of the Rho GTPase-activating protein RICS in neurite outgrowth. AB - The Rho family of small GTPases, including RhoA, Rac1 and Cdc42, are critical regulators of the actin cytoskeleton. In neuronal systems, Rho GTPase-activating proteins (RhoGAPs) and their substrates, Rho GTPases, have been implicated in regulating multiple processes in the morphological development of neurons, including axonal growth and guidance, dendritic elaboration and formation of synapses. RICS is mainly expressed in the brain and functions as a RhoGAP protein for Cdc42 and Rac1 in vitro. To examine the biological function of RICS, we disrupted the RICS gene in mice. RICS knockout mice developed normally and were fertile. However, when cultured in vitro, Cdc42 activity in RICS(-/-) neurons was higher than that in wild-type neurons. Consistent with this finding, hippocampal and cerebellar granule neurons derived from RICS(-/-) mice bore longer neurites than those from wild-type mice. These findings suggest that RICS plays an important role in neurite extension by regulating Cdc42 in vivo. PMID- 16716192 TI - Non conservation of the meiotic function of the Ski8/Rec103 homolog in Arabidopsis. AB - Meiotic recombination involves the formation and repair of DNA double-strand breaks (DSB). One of the genes required for DSB formation in the yeast Saccharomyces cerevisiae, Ski8/Rec103, is intriguing because it also plays a role in cytoplasmic RNA metabolism, a function difficult to relate to DSB formation. The meiotic role of Ski8 is conserved in several fungi, but has not been investigated outside this kingdom. We identified the Ski8 homolog in Arabidopsis thaliana and isolated two mutants. We showed that the Arabidopsis Ski8 homolog was required for normal plant development and growth, suggesting a conserved somatic function, but that it was not required for meiotic recombination or progression. The data presented here provide strong evidence that the meiotic role of Ski8 is not conserved in Arabidopsis and sequence analysis suggests that this may also be the case in a range of other species. PMID- 16716193 TI - Decreased basal mucus secretion by Slp2-a-deficient gastric surface mucous cells. AB - Synaptotagmin-like protein (Slp) 2-a is a putative Rab27A/B-effector protein and is implicated in intracellular membrane transport. However, the precise tissue distribution of Slp2-a protein and its functions remain largely unknown. In this study we used a specific anti-Slp2-a antibody to investigate the tissue distribution of Slp2-a in mice and found that Slp2-a is most abundantly expressed in mouse stomach. Co-immunoprecipitation experiments indicated that Slp2-a interacts with Rab27A/B in vivo. We also discovered that Slp2-a and Rab27A/B are predominantly localized at the apical region of gastric-surface mucous cells, where mucus granules are accumulated. Analysis of Slp2-a mutant mice generated by homologous recombination showed a reduced number of mucus granules, a deficiency of granule docking with the apical plasma membrane in the gastric-surface mucous cells and reduction of mucus secretion by Slp2-a-deficient gastric primary cells. Based on these results, we propose that Slp2-a is part of the mucin secretory machinery in surface mucous cells of mouse stomach. PMID- 16716194 TI - Role of APP phosphorylation in FE65-dependent gene transactivation mediated by AICD. AB - Consecutive cleavages of Alzheimer's amyloid beta-protein precursor (APP) generate intracellular domain fragment (AICD). Interaction of APP and/or AICD with the adaptor protein FE65 is thought to modulate the metabolism of APP and the function of AICD. Phosphorylation or amino acid substitution of APP and AICD at threonine 668 (Thr668) suppresses their association with FE65. Here, we analyzed the function of APP and AICD phosphorylation in the nuclear translocation of FE65. In brain, AICD was present as phosphorylated and non phosphorylated forms with non-phosphorylated AICD being dominantly detected in the nucleus. However, a mutant AICD (AICDa), in which Thr668 of AICD was replaced with Ala, was also mostly localized to the nucleus. These observations indicate that phosphorylation of AICD does not regulate the translocation of FE65 and that FE65 does not accompany AICD into the nucleus. APP was known to tether FE65 to the membrane. We found that phosphorylation of APP liberated membrane-bound FE65, which was then translocated into the nucleus where it up-regulated gene transactivation mediated by AICD, which was translocated into the nucleus independently of FE65. Therefore, phosphorylation of APP but not AICD modulates FE65-dependent gene transactivation mediated by AICD through the regulation of FE65 intracellular localization. PMID- 16716195 TI - The four members of the Drosophila metallothionein family exhibit distinct yet overlapping roles in heavy metal homeostasis and detoxification. AB - Four metallothionein genes are present in the Drosophila melanogaster genome, designated MtnA, MtnB, MtnC, MtnD, all of which are transcriptionally induced by heavy metals through the same metal-responsive transcription factor, MTF-1. Here we show, by targeted mutagenesis, that the four metallothionein genes exhibit distinct, yet overlapping, roles in heavy metal homeostasis and toxicity prevention. Among the individual metallothionein mutants, the most prominent distinction between them was that MtnA-defective flies were the most sensitive to copper load, while MtnB-defective flies were the most sensitive to cadmium. Using various reporter gene constructs and mRNA quantification, we show that the MtnA promoter is preferentially induced by copper, while the MtnB promoter is preferentially induced by cadmium. Such a metal preference is also observed at the protein level as the stoichiometric, spectrometric and spectroscopic features of the copper and cadmium complexes with MtnA and MtnB correlate well with a greater stability of copper-MtnA and cadmium-MtnB. Finally, MtnC and MtnD, both of which are very similar to MtnB, display lower copper and cadmium binding capabilities compared to either MtnA or MtnB. In accordance with these binding studies, Drosophila mutants of MtnC or MtnD have a near wild type level of resistance against copper or cadmium load. Furthermore, eye-specific over expression of MtnA and MtnB, but not of MtnC or MtnD, can rescue a "rough eye" phenotype caused by copper load in the eye. Taken together, while the exact roles of MtnC and MtnD remain to be determined, the preferential protection against copper and cadmium toxicity by MtnA and MtnB, respectively, are the result of a combination of promoter preference and metal binding. PMID- 16716196 TI - ELKS, a protein structurally related to the active zone protein CAST, is involved in Ca2+-dependent exocytosis from PC12 cells. AB - The active zone protein CAST binds directly to the other active zone proteins RIM, Bassoon and Piccolo, and it has been suggested that these protein-protein interactions play an important role in neurotransmitter release. To further elucidate the molecular mechanism, we attempted to examine the function of CAST using PC12 cells as a model system. Although PC12 cells do not express CAST, they do express ELKS, a protein structurally related to CAST. Endogenous and exogenously expressed ELKS, RIM2 and Bassoon were colocalized in punctate signals in PC12 cells. Over-expression of full-length ELKS resulted in a significant increase in stimulated exocytosis of human growth hormone (hGH) from PC12 cells, similar to the effect of full-length RIM2. This increase was not observed following over-expression of deletion constructs of ELKS that lacked either the last three amino acids (IWA) required for binding to RIM2 or a central region necessary for binding to Bassoon. Moreover, over-expression of the NH(2)-terminal RIM2-binding domain of Munc13-1, which is known to inhibit the binding between RIM and Munc13-1, inhibited the stimulated increase in hGH secretion by full length RIM2. Furthermore, this construct also inhibited the stimulated increase in hGH secretion induced by full-length ELKS. These results suggest that ELKS is involved in Ca(2+)-dependent exocytosis from PC12 cells at least partly via the RIM2-Munc13-1 pathway. PMID- 16716197 TI - Comprehensive analysis of the ICEN (Interphase Centromere Complex) components enriched in the CENP-A chromatin of human cells. AB - The centromere is a chromatin structure essential for correct segregation of sister chromatids, and defects in this region often lead to aneuploidy and cancer. We have previously reported purification of the interphase centromere complex (ICEN) from HeLa cells, and have demonstrated the presence of 40 proteins (ICEN1-40), along with CENP-A, -B, -C, -H and hMis6, by proteomic analysis. Here we report analysis of seven ICEN components with unknown function. Centromere localization of EGFP-tagged ICEN22, 24, 32, 33, 36, 37 and 39 was observed in transformant cells. Depletion of each of these proteins by short RNA interference produced abnormal metaphase cells carrying misaligned chromosomes and also produced cells containing aneuploid chromosomes, implying that these ICEN proteins take part in kinetochore functions. Interestingly, in the ICEN22, 32, 33, 37 or 39 siRNA-transfected cells, CENP-H and hMis6 signals disappeared from all the centromeres in abnormal mitotic cells containing misaligned chromosomes. These results suggest that the seven components of the ICEN complex are predominantly localized at the centromeres and are required for kinetochore function perhaps through or not through loading of CENP-H and hMis6 onto the centromere. PMID- 16716198 TI - Transgenic rescue of erythroid 5-aminolevulinate synthase-deficient mice results in the formation of ring sideroblasts and siderocytes. AB - Molecular defects in erythroid 5-aminolevulinate synthase (ALAS-E), the first enzyme in the heme biosynthetic pathway, cause X-linked sideroblastic anemia (XLSA). However, ring sideroblasts, the hallmark of XLSA, were not found in ALAS E-deficient mouse embryos, indicating that simple ALAS-E-deficiency is not sufficient for ring sideroblast formation. To investigate the developmental stage specific pathogenesis caused by heme-depletion, we attempted a complementation rescue of ALAS-E-deficiency. We exploited transgenic mouse lines expressing human ALAS-E at approximately half that of wild-type levels. In these hypomorphic embryos, most of the primitive erythroid cells were transformed into ring sideroblasts. The majority of the circulating definitive erythroid cells became siderocytes, enucleated erythrocytes containing iron deposits, and definitive ring sideroblasts were also observed. These iron-overloaded cells suffered from an alpha/beta globin chain imbalance. Despite the iron overload, transferrin receptors were highly expressed in the erythroid cells, suggesting they contribute to the formation of ring sideroblasts and siderocytes. These results indicate that a partially depleted heme supply provokes ring sideroblast formation. The experimental generation of ring sideroblasts in animals would contribute to our understanding of the iron metabolism and its disorder in erythroid cells. PMID- 16716200 TI - The mechanism of emergenesis. AB - Since each individual produced by the sexual process contains a unique set of genes, very exceptional combinations of genes are unlikely to appear twice even within the same family. E. O. Wilson (1978)The intraclass correlations of monozygotic twins who were separated in infancy and reared apart (MZA twins) provide estimates of trait heritability, and the Minnesota Study of Twins Reared Apart [MISTRA: Bouchard et al. (1990), The sources of human psychological differences: the Minnesota study of twins reared apart, Science 250, 223-228] has demonstrated that MZA pairs are as similar in most respects as MZ pairs reared together. Some polygenic traits--e.g. stature, IQ, harm avoidance, negative emotionality, interest in sports--are polygenic-additive, so pairs of relatives resemble one another on the given trait in proportion to their genetic similarity. But the existence and the intensity of other important psychological traits seem to be emergent properties of gene configurations (or configurations of independent and partially genetic traits) that interact multiplicatively rather than additively. Monozygotic (MZ) twins may be strongly correlated on such emergenic traits, while the similarity of dizygotic (DZ) twins, sibs or parent offspring pairs may be much less than half that of MZ pairs. Some emergenic traits, although strongly genetic, do not appear to run in families. MISTRA has provided at least two examples of traits for which MZA twins are strongly correlated, and DZA pairs correlate near zero, while DZ pairs reared together (DZTs) are about half as similar as MZTs. These findings suggest that even more traits may be emergenic than those already identified. Studies of adoptees reared together (who are perhaps more common than twins reared apart) may help to identify traits that are emergenic, but that also are influenced by a common rearing environment. PMID- 16716201 TI - The molecular genetics of cognition: dopamine, COMT and BDNF. AB - The important contribution of genetic factors to the development of cognition and intelligence is widely acknowledged, but identification of these genes has proven to be difficult. Given a variety of evidence implicating the prefrontal cortex and its dopaminergic circuits in cognition, most of the research conducted to date has focused on genes regulating dopaminergic function. Here we review the genetic association studies carried out on catechol-O-methyltransferase (COMT) and the dopamine receptor genes, D1, D2 and D4. In addition, the evidence implicating another promising candidate gene, brain-derived neurotrophic factor (BDNF) in neuropsychological function, is assessed. Both the COMT val158met polymorphism and the BDNF val66met variant appear to influence cognitive function, but the specific neurocognitive processes involved continue to be a matter of debate. Part of the difficulty is distinguishing between false positives, pleiotropy and the influence of a general intelligence factor, g. Also at issue is the complexity of the relevant neuromolecular pathways, which make the inference of simple causal relationships difficult. The implications of molecular genetic cognitive research for psychiatry are discussed in light of these data. PMID- 16716202 TI - Tissue-specific alternative splicing of BK channel transcripts in Drosophila. AB - BK-type calcium-activated potassium channels are large conductance channels that respond to changes in intracellular calcium and membrane potential. These channels are used in a wide variety of cell types and have recently been linked to drug sensitivity and tolerance. In both Drosophila and mammals, BK channels are encoded by the slowpoke gene. The Drosophila slowpoke gene includes 14 alternative exons distributed among five sites of alternative splicing. Presumably, the purpose of alternative processing is to provide transcripts tailored to the needs of the cell. The slowpoke gene is expressed in nervous, muscle and epithelial tissues. To determine whether splicing is controlled in a tissue- and/or developmental-specific manner, we built tissue- and developmental specific cDNA libraries that preserved the relative frequency of various slowpoke splice variants. These libraries were screened by colony hybridization using alternative exon-specific DNA probes to document the frequency of individual alternative exons in different developmental stages and distinct tissue types. We demonstrate that slowpoke transcripts undergo tissue- and developmental-specific splicing in Drosophila and some exons are diagnostic for specific tissues. PMID- 16716203 TI - Investigation of A218C tryptophan hydroxylase polymorphism: association with familial suicide behavior and proband's suicide attempt characteristics. AB - According to WHO, suicide accounts for about 1,000,000 deaths worldwide every year. In view of these dramatic data, several studies have tried to identify possible biological mechanisms and markers of suicide. Genes encoding for proteins involved in the serotonergic transmission are major candidates in association studies of suicidal behavior. The gene that codes for tryptophan hydroxylase (TPH), the rate-limiting enzyme in the biosynthesis of serotonin, is one of these candidates. Two polymorphisms in intron 7 of this gene (A218C and A779C) have been described, but their role in suicidal behavior remains uncertain. TPH A218C polymorphism was analyzed in a sample of 248 psychiatric patients and 63 healthy controls. In addition, at least one close relative member was interviewed to assess family suicidal behavior history. Our research confirmed that a positive history of suicide attempts in a family member is associated with the chance of an individual to attempt suicide. Furthermore, we demonstrated that familial suicide attempts are more lethal and frequently more violent. We were not able to find significant differences of the TPH genotype frequencies between patients and controls. The TPH A218C genotypes were not associated with a history of suicide attempt and the lethality of the most lethal lifetime suicide attempt and suicide attempt method. The authors conclude that the A218C polymorphism of the TPH gene may not be a susceptibility factor for suicidal behavior in this group of psychiatric patients but confirm that a family suicidal behavior history increases the proband's suicide attempt risk. PMID- 16716204 TI - Effects of repeated maternal separation on prepulse inhibition of startle across inbred mouse strains. AB - A growing body of research implicates genetic factors and childhood trauma in the etiology of neuropsychiatric diseases such as schizophrenia. However, there remains little understanding of how genetic variation influences early life stress to affect later disease susceptibility. Studies in rats have shown that postnatal maternal separation (MS) results in later deficits in prepulse inhibition of the acoustic startle response (PPI), an impairment in sensorimotor gating found in schizophrenic patients. In the present study, genetic differences in the effects of repeated MS on PPI were examined in eight inbred strains of mice (129S1/SvImJ, 129P3/J, A/J, BALB/cJ, BALB/cByJ C57BL/6J, DBA/2J and FVB/NJ). Mice were assigned to either MS (180 min/day on postnatal days P0-P13), 'handling' (15 min/day, P0-P13) or facility-reared conditions and tested for PPI at 12 weeks of age. Results demonstrated major strain differences in the production of viable offspring irrespective of MS, leading to the exclusion of 129P3/J, A/J and BALB/cJ from the study. Pups from the five remaining strains exhibited marked differences in the acoustic startle response and PPI, confirming previous strain comparisons. However, MS produced no significant effects on PPI in any of the strains tested. A second form of postnatal stress (repeated footshock) also failed to alter PPI in the one strain studied, C57BL/6J. Present results demonstrate that the form of MS studied herein does not provide a robust model of early life stress effects on PPI in the mouse strains tested. The development and validation of a reliable mouse model of early life stress remains an important research goal. PMID- 16716205 TI - Holeboard discrimination learning in mice. AB - We have adapted to mice a holeboard-learning task, which allows simultaneous assessment of spatial working and reference-memory performance. The holeboard apparatus consists of an open-field chamber with a 16-hole floor insert. Across trials, animals have to learn that the same four holes of 16 are always baited. Here, we show that C57BL/6 mice readily acquire this task within 4 days when submitted to six trials per day or within 8 days when submitted to only four trials per day. We also show that C57BL/6, Swiss-Webster, CD-1 and DBA/2 mice acquire this task similarly, despite the fact that some differences could be observed in measures of exploratory activity during habituation and training. Moreover, the muscarinic antagonist scopolamine disrupts learning at doses of 0.1 and 1.0 mg/kg, although the highest dose appeared to have side-effects. Lastly, we found that amyloid precursor protein transgenic mice have a selective disruption in their working-memory performance only during reversal training (i.e. after a change in the configuration of the baited holes). Overall, our data indicate that this spatial learning task is well adapted to mice and will be useful to characterize spatial memory in various genetic or pharmacological mouse models. PMID- 16716206 TI - Association study between kynurenine 3-monooxygenase gene and schizophrenia in the Japanese population. AB - Several lines of evidence suggest that metabolic changes in the kynurenic acid (KYNA) pathway are related to the etiology of schizophrenia. The inhibitor of kynurenine 3-monooxygenase (KMO) is known to increase KYNA levels, and the KMO gene is located in the chromosome region associated with schizophrenia, 1q42-q44. Single-marker and haplotype analyses for 6-tag single nucleotide polymorphisms (SNPs) of KMO were performed (cases = 465, controls = 440). Significant association of rs2275163 with schizophrenia was observed by single-marker comparisons (P = 0.032) and haplotype analysis including this SNP (P = 0.0049). Significant association of rs2275163 and haplotype was not replicated using a second, independent set of samples (cases = 480, controls = 448) (P = 0.706 and P = 0.689, respectively). These results suggest that the KMO is unlikely to be related to the development of schizophrenia in Japanese. PMID- 16716207 TI - Phenotype-genotype association grid: a convenient method for summarizing multiple association analyses. AB - BACKGROUND: High-throughput genotyping generates vast amounts of data for analysis; results can be difficult to summarize succinctly. A single project may involve genotyping many genes with multiple variants per gene and analyzing each variant in relation to numerous phenotypes, using several genetic models and population subgroups. Hundreds of statistical tests may be performed for a single SNP, thereby complicating interpretation of results and inhibiting identification of patterns of association. RESULTS: To facilitate visual display and summary of large numbers of association tests of genetic loci with multiple phenotypes, we developed a Phenotype-Genotype Association (PGA) grid display. A database-backed web server was used to create PGA grids from phenotypic and genotypic data (sample sizes, means and standard errors, P-value for association). HTML pages were generated using Tcl scripts on an AOLserver platform, using an Oracle database, and the ArsDigita Community System web toolkit. The grids are interactive and permit display of summary data for individual cells by a mouse click (i.e. least squares means for a given SNP and phenotype, specified genetic model and study sample). PGA grids can be used to visually summarize results of individual SNP associations, gene-environment associations, or haplotype associations. CONCLUSION: The PGA grid, which permits interactive exploration of large numbers of association test results, can serve as an easily adapted common and useful display format for large-scale genetic studies. Doing so would reduce the problem of publication bias, and would simplify the task of summarizing large scale association studies. PMID- 16716208 TI - Using the Ages and Stages Questionnaire to teach medical students developmental assessment: a descriptive analysis. AB - BACKGROUND: After a survey of medical graduates' skills found a lack of confidence in developmental assessment, a program was introduced with the broad aims of increasing medical student confidence and respect for the parents' role in childhood developmental assessment. Research has shown that parents' concerns are as accurate as quality screening tests in assessing development, so the program utilised the Ages and Stages Questionnaire, a parent completed, child development assessment tool. METHOD: To evaluate the program, an interpretative analysis was completed on the students' reports written during the program and a questionnaire was administered to the parents to gain their perception of the experience. As well, student confidence levels in assessing growth and development were measured at the end of the paediatric term. RESULTS: Although there was an increase in student confidence in developmental assessment at the end of the term, it was not statistically significant. However the findings indicated that students gained increased understanding of the process and enhanced recognition of the parental role, and the study suggested there was increased confidence in some students. Parents indicated that they thought they should be involved in the teaching of students. CONCLUSION: The ASQ was shown to have been useful in an education program at the level of advanced beginners in developmental assessment. PMID- 16716210 TI - Topiramate plus citalopram in the treatment of Compulsive-Impulsive Sexual Behaviors. AB - Compulsive-Impulsive Sexual Behaviors (C-ISBs) include repetitive sexual acts and compulsive sexual thoughts which occur so frequently and with such intensity that they interfere with sexual intimacy and interpersonal and occupational functioning and whose categorization and effective treatments are still unclear. We report the case of a patient affected by C-ISBs and bipolar disorder of type II who improved dramatically after three months' addition of topiramate to citalopram. Topiramate is a powerful anticonvulsant which has recently been proposed also for the treatment of migraine, bipolar disorder and binge eating disorder. This case-report suggests that topiramate might be beneficial in augmentation with citalopram in patients suffering from C-ISBs, although controlled studies to confirm our findings are needed. PMID- 16716209 TI - Widespread ectopic expression of olfactory receptor genes. AB - BACKGROUND: Olfactory receptors (ORs) are the largest gene family in the human genome. Although they are expected to be expressed specifically in olfactory tissues, some ectopic expression has been reported, with special emphasis on sperm and testis. The present study systematically explores the expression patterns of OR genes in a large number of tissues and assesses the potential functional implication of such ectopic expression. RESULTS: We analyzed the expression of hundreds of human and mouse OR transcripts, via EST and microarray data, in several dozens of human and mouse tissues. Different tissues had specific, relatively small OR gene subsets which had particularly high expression levels. In testis, average expression was not particularly high, and very few highly expressed genes were found, none corresponding to ORs previously implicated in sperm chemotaxis. Higher expression levels were more common for genes with a non-OR genomic neighbor. Importantly, no correlation in expression levels was detected for human-mouse orthologous pairs. Also, no significant difference in expression levels was seen between intact and pseudogenized ORs, except for the pseudogenes of subfamily 7E which has undergone a human-specific expansion. CONCLUSION: The OR superfamily as a whole, show widespread, locus dependent and heterogeneous expression, in agreement with a neutral or near neutral evolutionary model for transcription control. These results cannot reject the possibility that small OR subsets might play functional roles in different tissues, however considerable care should be exerted when offering a functional interpretation for ectopic OR expression based only on transcription information. PMID- 16716212 TI - Improvement of bioprocess monitoring: development of novel concepts. AB - The advancement of bioprocess monitoring will play a crucial role to meet the future requirements of bioprocess technology. Major issues are the acceleration of process development to reduce the time to the market and to ensure optimal exploitation of the cell factory and further to cope with the requirements of the Process Analytical Technology initiative. Due to the enormous complexity of cellular systems and lack of appropriate sensor systems microbial production processes are still poorly understood. This holds generally true for the most microbial production processes, in particular for the recombinant protein production due to strong interaction between recombinant gene expression and host cell metabolism. Therefore, it is necessary to scrutinise the role of the different cellular compartments in the biosynthesis process in order to develop comprehensive process monitoring concepts by involving the most significant process variables and their interconnections. Although research for the development of novel sensor systems is progressing their applicability in bioprocessing is very limited with respect to on-line and in-situ measurement due to specific requirements of aseptic conditions, high number of analytes, drift, and often rather low physiological relevance. A comprehensive survey of the state of the art of bioprocess monitoring reveals that only a limited number of metabolic variables show a close correlation to the currently explored chemical/physical principles. In order to circumvent this unsatisfying situation mathematical methods are applied to uncover "hidden" information contained in the on-line data and thereby creating correlations to the multitude of highly specific biochemical off-line data. Modelling enables the continuous prediction of otherwise discrete off-line data whereby critical process states can be more easily detected. The challenging issue of this concept is to establish significant on-line and off-line data sets. In this context, online sensor systems are reviewed with respect to commercial availability in combination with the suitability of offline analytical measurement methods. In a case study, the aptitude of the concept to exploit easily available online data for prediction of complex process variables in a recombinant E. coli fed-batch cultivation aiming at the improvement of monitoring capabilities is demonstrated. In addition, the perspectives for model-based process supervision and process control are outlined. PMID- 16716211 TI - The effect of a worksite based walking programme on cardiovascular risk in previously sedentary civil servants [NCT00284479]. AB - BACKGROUND: A significant proportion of Europeans do not meet the recommendations for 30 mins of physical activity 5 times per week. Whether lower frequency, moderate intensity exercise alters cardiovascular disease (CVD) risk has received little attention. This study examined the effects of 45 minutes self-paced walking, 2 d. wk(-1) on aerobic fitness, blood pressure (BP), body composition, lipids and C-Reactive Protein (CRP) in previously sedentary civil servants. METHODS: 37 subjects (24 women) aged 41.5 +/- 9.3 years were randomly assigned to either two 45 minute walks per week (walking group) or no training (control group). Aerobic fitness, body composition, blood pressure (BP), CRP and lipoprotein variables were measured at baseline and following 8 weeks. Steps counts were measured at baseline and during weeks 4 and 8 of the intervention. RESULTS: Compared to the control group, the walking group showed a significant reduction in systolic BP and maintained body fat levels (P < 0.05). There were no changes other risk factors. Subjects took significantly more steps on the days when prescribed walking was performed (9303 +/- 2665) compared to rest days (5803 +/- 2749; P < 0.001). CONCLUSION: These findings suggest that walking twice per week for 45 minutes at approximately 62% HRmax, improves activity levels, reduces systolic BP and prevents an increase in body fat in previously sedentary adults. This walking prescription, however, failed to induce significant improvements in other markers of cardiovascular disease risk following eight weeks of training. PMID- 16716213 TI - Identification of a Bacillus thuringiensis Cry11Ba toxin-binding aminopeptidase from the mosquito, Anopheles quadrimaculatus. AB - BACKGROUND: Aminopeptidase N (APN) type proteins isolated from several species of lepidopteran insects have been implicated as Bacillus thuringiensis (Bt) toxin binding proteins (receptors) for Cry toxins. We examined brush border membrane vesicle (BBMV) proteins from the mosquito Anopheles quadrimaculatus to determine if APNs from this organism would bind mosquitocidal Cry toxins that are active to it. RESULTS: A 100-kDa protein with APN activity (APNAnq 100) was isolated from the brush border membrane of Anopheles quadrimaculatus. Native state binding analysis by surface plasmon resonance shows that APNAnq 100 forms tight binding to a mosquitocidal Bt toxin, Cry11Ba, but not to Cry2Aa, Cry4Ba or Cry11Aa. CONCLUSION: An aminopeptidase from Anopheles quadrimaculatus mosquitoes is a specific binding protein for Bacillus thuringiensis Cry11Ba. PMID- 16716214 TI - Biological efficacy of low versus medium dose aspirin after coronary surgery: results from a randomized trial [NCT00262275]. AB - BACKGROUND: The beneficial effect of aspirin after coronary surgery is established; however, a recent study reported the inability of low doses (100 mg) to inhibit postoperative platelet function. We conducted a double-blind randomised trial to establish the efficacy of low dose aspirin and to compare it against medium dose aspirin. METHODS: Patients undergoing coronary surgery were invited to participate and consenting patients were randomised to 100 mg or 325 mg of aspirin daily for 5 days. Our primary outcome was the difference in platelet aggregation (day 5 - baseline) using 1 microg/ml of collagen. Secondary outcomes were differences in EC50 of collagen, ADP and epinephrine (assessed using the technique of Born). RESULTS: From September 2002 to April 2004, 72 patients were randomised; 3 patients discontinued, leaving 35 and 34 in the low and medium dose aspirin arms respectively. The mean aggregation (using 1.1 microg/ml of collagen) was reduced in both the medium and low dose aspirin arms by 37% and 36% respectively. The baseline adjusted difference (low - medium) was 6% (95% CI -3 to 14; p = 0.19). The directions of the results for the differences in EC50 (low - medium) were consistent for collagen, ADP and epinephrine at -0.07 (-0.53 to 0.40), -0.08 (-0.28 to 0.11) and -4.41 (-10.56 to 1.72) respectively, but none were statistically significant. CONCLUSION: Contrary to recent findings, low dose aspirin is effective and medium dose aspirin did not prove superior for inhibiting platelet aggregation after coronary surgery. PMID- 16716216 TI - Self-harm in British South Asian women: psychosocial correlates and strategies for prevention. AB - OBJECTIVE: To review the rates of self-harm in British South Asian women, look into the factors that contribute to these high rates of self-harm and discuss possible strategies for prevention and provision of culturally sensitive service for South Asian women who harm themselves. METHOD: Review. RESULTS: South Asian women are significantly more likely to self harm between ages 16-24 years than white women. Across all age groups the rates of self harm are lower in South Asian men as compared to South Asian women. These women are generally younger, likely to be married and less likely to be unemployed or use alcohol or other drugs. They report more relationship problems within the family. South Asian women are less likely to attend the ER with repeat episode since they hold the view that mainstream services do not meet their needs. CONCLUSION: South Asian women are at an increased risk of self harm. Their demographic characteristics, precipitating factors and clinical management are different than whites. There is an urgent need for all those concerned with the mental health services for ethnic minorities to take positive action and eradicate the barriers that prevent British South Asians from seeking help. There is a need to move away from stereotypes and overgeneralisations and start from the user's frame of reference, taking into account family dynamics, belief systems and cultural constraints. PMID- 16716215 TI - Spatial normalization of array-CGH data. AB - BACKGROUND: Array-based comparative genomic hybridization (array-CGH) is a recently developed technique for analyzing changes in DNA copy number. As in all microarray analyses, normalization is required to correct for experimental artifacts while preserving the true biological signal. We investigated various sources of systematic variation in array-CGH data and identified two distinct types of spatial effect of no biological relevance as the predominant experimental artifacts: continuous spatial gradients and local spatial bias. Local spatial bias affects a large proportion of arrays, and has not previously been considered in array-CGH experiments. RESULTS: We show that existing normalization techniques do not correct these spatial effects properly. We therefore developed an automatic method for the spatial normalization of array CGH data. This method makes it possible to delineate and to eliminate and/or correct areas affected by spatial bias. It is based on the combination of a spatial segmentation algorithm called NEM (Neighborhood Expectation Maximization) and spatial trend estimation. We defined quality criteria for array-CGH data, demonstrating significant improvements in data quality with our method for three data sets coming from two different platforms (198, 175 and 26 BAC-arrays). CONCLUSION: We have designed an automatic algorithm for the spatial normalization of BAC CGH-array data, preventing the misinterpretation of experimental artifacts as biologically relevant outliers in the genomic profile. This algorithm is implemented in the R package MANOR (Micro-Array NORmalization), which is described at http://bioinfo.curie.fr/projects/manor and available from the Bioconductor site http://www.bioconductor.org. It can also be tested on the CAPweb bioinformatics platform at http://bioinfo.curie.fr/CAPweb. PMID- 16716217 TI - Measuring inequality: tools and an illustration. AB - BACKGROUND: This paper examines an aspect of the problem of measuring inequality in health services. The measures that are commonly applied can be misleading because such measures obscure the difficulty in obtaining a complete ranking of distributions. The nature of the social welfare function underlying these measures is important. The overall object is to demonstrate that varying implications for the welfare of society result from inequality measures. METHOD: Various tools for measuring a distribution are applied to some illustrative data on four distributions about mental health services. Although these data refer to this one aspect of health, the exercise is of broader relevance than mental health. The summary measures of dispersion conventionally used in empirical work are applied to the data here, such as the standard deviation, the coefficient of variation, the relative mean deviation and the Gini coefficient. Other, less commonly used measures also are applied, such as Theil's Index of Entropy, Atkinson's Measure (using two differing assumptions about the inequality aversion parameter). Lorenz curves are also drawn for these distributions. RESULTS: Distributions are shown to have differing rankings (in terms of which is more equal than another), depending on which measure is applied. CONCLUSION: The scope and content of the literature from the past decade about health inequalities and inequities suggest that the economic literature from the past 100 years about inequality and inequity may have been overlooked, generally speaking, in the health inequalities and inequity literature. An understanding of economic theory and economic method, partly introduced in this article, is helpful in analysing health inequality and inequity. PMID- 16716218 TI - Use of complementary and alternative medicine in Germany - a survey of patients with inflammatory bowel disease. AB - BACKGROUND: Previous studies have suggested an increasing use of complementary and alternative medicine (CAM) in patients with inflammatory bowel disease (IBD). The aim of our study was to evaluate the use of CAM in German patients with IBD. METHODS: A questionnaire was offered to IBD patients participating in patient workshops which were organized by a self-help association, the German Crohn's and Colitis Association. The self-administered questionnaire included demographic and disease-related data as well as items analysing the extent of CAM use and satisfaction with CAM treatment. Seven commonly used CAM methods were predetermined on the questionnaire. RESULTS: 413 questionnaires were completed and included in the analysis (n = 153 male, n = 260 female; n = 246 Crohn's disease, n = 164 ulcerative colitis). 52 % of the patients reported CAM use in the present or past. In detail, homeopathy (55%), probiotics (43%), classical naturopathy (38%), Boswellia serrata extracts (36%) and acupuncture/Traditional Chinese Medicine (TCM) (33%) were the most frequently used CAM methods. Patients using probiotics, acupuncture and Boswellia serrata extracts (incense) reported more positive therapeutic effects than others. Within the statistical analysis no significant predictors for CAM use were found. 77% of the patients felt insufficiently informed about CAM. CONCLUSION: The use of CAM in IBD patients is very common in Germany, although a large proportion of patients felt that information about CAM is not sufficient. However, to provide an evidence-based approach more research in this field is desperately needed. Therefore, physicians should increasingly inform IBD patients about benefits and limitations of CAM treatment. PMID- 16716219 TI - Tissue localization of collagenase and leucine aminopeptidase in the bovine filarial parasite Setaria cervi. AB - BACKGROUND: Like other helminth proteases, filarial proteases have also been shown to require for parasite survival inside the host and mediate various physiologic processes such as tissue invasion, feeding, embryogenesis and host immune evasion. Many of these proteases have shown potential for vaccines and chemotherapeutic agents against active filarial infections. Setaria cervi is a bovine filarial parasite and serves as a good parasite model for the studies in lymphatic filariasis. Recently, a 175 kDa collagenase and leucine aminopeptidase (LAP) have been purified and characterized from the bovine filarial parasite S. cervi and shown to be potential vaccine candidate and diagnostic marker, respectively for human lymphatic filariasis. However, their tissue localizations and putative roles in the parasite biology have not yet been examined and thus remain unclear. Therefore, the current study attempts to localize and explore the putative roles of these two enzymes in S. cervi. METHODS: The tissue distributions of 175 kDa collagenase and leucine aminopeptidase in S. cervi were examined by immunohistochemical and histochemical methods, respectively. Immune sera obtained from the jirds immunized with collagenase served as primary antibody, rabbit anti-mouse IgG-HRP conjugate as secondary antibody and DAB as the substrate for the immunostaining of collagenase. Leu-betaNA was used as the substrate for the histochemical staining of LAP. RESULTS: Both the collagenase and LAP were present in the body wall; however, they differ in their distribution pattern in different layers of body wall. Collagenase was mainly localized in epicuticle, cuticle, syncytial hypodermis and the nerve cord region whereas LAP was more concentrated in epicuticle, longitudinal muscle layers and almost absent or very faintly stained in syncytial hypodermis and nerve cord region. Both collagenase and LAP showed their common distributions in intestine, uterus and mature eggs, growing embryos and mf. Very strong immunostaining of collagenase in the outer body surface of the parasite indicates its major role in host-parasite relationship whereas the presence of LAP in muscular region suggests its role in tissue remodeling. The common presences of collagenase and LAP in the S. cervi intestine, ovary, uterus, eggs and mf suggest that they also have collaborative roles in molting, nutrition and embryogenesis. The data obtained on their immunological characterizations and their presence in important parasite organs give strong indication that they are critical for the survival of filarial parasite and thus can be good vaccine candidates and/or diagnostic markers for human lymphatic filariasis. CONCLUSION: The manuscript reports for the first time the tissue distribution of collagenase and LAP in the bovine filarial parasite S. cervi and discuss their putative roles in vivo. Our findings also open the avenue to examine the roles of these two proteases in vivo, which will require further experiments like using their natural substrates and/or specific inhibitors in each tissues. PMID- 16716221 TI - Gene by environment QTL mapping through multiple trait analyses in blood pressure salt-sensitivity: identification of a novel QTL in rat chromosome 5. AB - BACKGROUND: The genetic mechanisms underlying interindividual blood pressure variation reflect the complex interplay of both genetic and environmental variables. The current standard statistical methods for detecting genes involved in the regulation mechanisms of complex traits are based on univariate analysis. Few studies have focused on the search for and understanding of quantitative trait loci responsible for gene x environmental interactions or multiple trait analysis. Composite interval mapping has been extended to multiple traits and may be an interesting approach to such a problem. METHODS: We used multiple-trait analysis for quantitative trait locus mapping of loci having different effects on systolic blood pressure with NaCl exposure. Animals studied were 188 rats, the progenies of an F2 rat intercross between the hypertensive and normotensive strain, genotyped in 179 polymorphic markers across the rat genome. To accommodate the correlational structure from measurements taken in the same animals, we applied univariate and multivariate strategies for analyzing the data. RESULTS: We detected a new quantitative train locus on a region close to marker R589 in chromosome 5 of the rat genome, not previously identified through serial analysis of individual traits. In addition, we were able to justify analytically the parametric restrictions in terms of regression coefficients responsible for the gain in precision with the adopted analytical approach. CONCLUSION: Future work should focus on fine mapping and the identification of the causative variant responsible for this quantitative trait locus signal. The multivariable strategy might be valuable in the study of genetic determinants of interindividual variation of antihypertensive drug effectiveness. PMID- 16716220 TI - Outer membrane protein genes and their small non-coding RNA regulator genes in Photorhabdus luminescens. AB - INTRODUCTION: Three major outer membrane protein genes of Escherichia coli, ompF, ompC, and ompA respond to stress factors. Transcripts from these genes are regulated by the small non-coding RNAs micF, micC, and micA, respectively. Here we examine Photorhabdus luminescens, an organism that has a different habitat from E. coli for outer membrane protein genes and their regulatory RNA genes. RESULTS: By bioinformatics analysis of conserved genetic loci, mRNA 5'UTR sequences, RNA secondary structure motifs, upstream promoter regions and protein sequence homologies, an ompF -like porin gene in P. luminescens as well as a duplication of this gene have been predicted. Gene loci for micF RNA, as well as OmpC protein and its associated regulatory micC RNA, were not found. Significantly, a sequence bearing the appropriate signatures of the E. coli micA RNA was located. The ompA homolog was previously annotated in P. luminescens. CONCLUSION: Presence of an ompF-like porin in P. luminescens is in keeping with the necessity to allow for passage of small molecules into the cell. The apparent lack of ompC, micC and micF suggests that these genes are not essential to P. luminescens and ompC and micF in particular may have been lost when the organism entered its defined life cycle and partially protected habitat. Control of porin gene expression by RNA may be more prevalent in free- living cells where survival is dependent on the ability to make rapid adjustments in response to environmental stress. Regulation of ompA by micA may have been retained due to a necessity for ompA control during one or both stages of the P. luminescens life cycle. PMID- 16716222 TI - Construction, purification, and characterization of a chimeric TH1 antagonist. AB - BACKGROUND: TH1 immune response antagonism is a desirable approach to mitigate some autoimmune and inflammatory reactions during the course of several diseases where IL-2 and IFN-gamma are two central players. Therefore, the neutralization of both cytokines could provide beneficial effects in patients suffering from autoimmune or inflammatory illnesses. RESULTS: A chimeric antagonist that can antagonize the action of TH1 immunity mediators, IFN-gamma and IL-2, was designed, engineered, expressed in E. coli, purified and evaluated for its in vitro biological activities. The TH1 antagonist molecule consists of the extracellular region for the human IFNgamma receptor chain 1 fused by a four aminoacid linker peptide to human 60 N-terminal aminoacid residues of IL-2. The corresponding gene fragments were isolated by RT-PCR and cloned in the pTPV-1 vector. E. coli (W3110 strain) was transformed with this vector. The chimeric protein was expressed at high level as inclusion bodies. The protein was partially purified by pelleting and washing. It was then solubilized with strong denaturant and finally refolded by gel filtration. In vitro biological activity of chimera was demonstrated by inhibition of IFN-gamma-dependent HLA-DR expression in Colo 205 cells, inhibition of IFN-gamma antiproliferative effect on HEp-2 cells, and by a bidirectional effect in assays for IL-2 T-cell dependent proliferation: agonism in the absence versus inhibition in the presence of IL-2. CONCLUSION: TH1 antagonist is a chimeric protein that inhibits the in vitro biological activities of human IFN-gamma, and is a partial agonist/antagonist of human IL-2. With these attributes, the chimera has the potential to offer a new opportunity for the treatment of autoimmune and inflammatory diseases. PMID- 16716223 TI - Smoking, respiratory symptoms and likely asthma in young people: evidence from postal questionnaire surveys in the Wythenshawe Community Asthma Project (WYCAP). AB - BACKGROUND: Although it is recognised that smoking is a major risk factor for subjects with chronic obstructive pulmonary disease and is associated with respiratory symptoms, there is less agreement concerning the relationship between asthma and smoking. This study aims to examine the relationship between cigarette smoking and asthma prevalence. METHOD: Data were used from two postal questionnaire surveys (1999 and 2001) in two general practice populations, using a respiratory questionnaire based on the ECRHQ and a generic quality of life questionnaire (EQ-5D). Only subjects less than 45 years old were included in the survey. An empirical definition of likely asthma was used based on respiratory questionnaire responses. Smoking was examined according to three categories, current smoker, ex smoker and never smoker. RESULTS: Almost 3500 subjects were included in the analyses. Current smokers had a higher prevalence of likely asthma compared to never smokers, odds ratio (OR) 1.59 (95% confidence interval (CI) 1.24 to 2.04). and also compared to ex smokers OR 1.79 (CI 1.25 to 2.56), but there was no difference between ex smokers and never smokers (OR 1.00 (0.75 1.35)). Current smoking was also positively associated with all symptoms but not with a history of hayfever/eczema. CONCLUSION: Although the positive association found between current smoking and obstructive airways disease is likely to be due to the effect of cigarettes on asthma, it could reflect an association with early COPD (GOLD stages 0 or 1). Smoking cessation has a beneficial effect on the prevalence of respiratory symptoms and is therefore of paramount importance among these young adults. PMID- 16716224 TI - Hannover study on long-stay hospitalization - part I: prediction of long-stay hospitalisation in cases of chronic mental illness. AB - BACKGROUND: The problem of long-stay hospitalization is still a pressing issue. In this study we examined the possibility of detecting and characterising the group at risk of long-stay hospitalization in advance. METHODS: This study examines the data of patients in the urban catchment area of the Medical University of Hannover, capital of Lower Saxony, Germany, during a period of 10 years. RESULTS AND CONCLUSION: The introduced "psychosocial risk-score", calculated at the first institutional contact, was able to predict the risk of long-term hospitalization. Characteristics of social disintegration, especially with regard to employment status, are of particular importance. PMID- 16716225 TI - Update: cohort mortality study of workers highly exposed to polychlorinated biphenyls (PCBs) during the manufacture of electrical capacitors, 1940-1998. AB - BACKGROUND: The National Institute for Occupational Safety and Health previously reported mortality for a cohort of workers considered highly exposed to polychlorinated biphenyls (PCBs) between 1939 and 1977 at two electrical capacitor manufacturing plants. The current study updated vital status, examined liver and rectal cancer mortality previously reported in excess in this cohort and evaluated mortality from non-Hodgkin's lymphoma (NHL) and cancers of the stomach, intestine, breast, prostate, skin (melanoma) and brain reported to be in excess in other cohort and case-control studies of PCB-exposed persons. METHODS: Mortality was updated through 1998 for 2572 workers. Age-, gender-, race- and calendar year-adjusted standardized mortality ratios (SMRs) and 95% confidence intervals (CI) were calculated using U.S., state and county referent rates. SMRs using U.S. referent rates are reported. Duration of employment was used as a surrogate for exposure. RESULTS: Consistent with the previous follow-up, mortality from biliary passage, liver and gall bladder cancer was significantly elevated (11 deaths, SMR 2.11, CI 1.05 - 3.77), but mortality from rectal cancer was not (6 deaths, SMR 1.47, CI 0.54 - 3.21). Among women, mortality from intestinal cancer (24 deaths, SMR 1.89, CI 1.21 - 2.82) and from "other diseases of the nervous system and sense organs", which include Parkinson's disease and amyotrophic lateral sclerosis, (15 deaths, SMR 2.07, CI 1.16 - 3.42) were elevated. There were four ALS deaths, all women (SMR 4.35, CI 1.19-11.14). Mortality was elevated for myeloma (7 deaths, SMR 2.11, CI 0.84 - 4.34), particularly among workers employed 10 years or more (5 deaths, SMR 2.80, CI 0.91 - 6.54). No linear associations between mortality and duration of employment were observed for the cancers of interest. CONCLUSION: This update found that the earlier reported excess in this cohort for biliary, liver and gall bladder cancer persisted with longer follow-up. Excess mortality for intestinal cancer among women was elevated across categories of duration of employment; myeloma mortality was highest among those working 10 years or more. The small numbers of deaths from liver and intestinal cancers, myeloma and nervous system diseases coupled with the lack of an exposure-response relationship with duration of employment preclude drawing definitive conclusions regarding PCB exposure and these causes of death. PMID- 16716226 TI - A jumping profile Hidden Markov Model and applications to recombination sites in HIV and HCV genomes. AB - BACKGROUND: Jumping alignments have recently been proposed as a strategy to search a given multiple sequence alignment A against a database. Instead of comparing a database sequence S to the multiple alignment or profile as a whole, S is compared and aligned to individual sequences from A. Within this alignment, S can jump between different sequences from A, so different parts of S can be aligned to different sequences from the input multiple alignment. This approach is particularly useful for dealing with recombination events. RESULTS: We developed a jumping profile Hidden Markov Model (jpHMM), a probabilistic generalization of the jumping-alignment approach. Given a partition of the aligned input sequence family into known sequence subtypes, our model can jump between states corresponding to these different subtypes, depending on which subtype is locally most similar to a database sequence. Jumps between different subtypes are indicative of intersubtype recombinations. We applied our method to a large set of genome sequences from human immunodeficiency virus (HIV) and hepatitis C virus (HCV) as well as to simulated recombined genome sequences. CONCLUSION: Our results demonstrate that jumps in our jumping profile HMM often correspond to recombination breakpoints; our approach can therefore be used to detect recombinations in genomic sequences. The recombination breakpoints identified by jpHMM were found to be significantly more accurate than breakpoints defined by traditional methods based on comparing single representative sequences. PMID- 16716227 TI - Transfection of oral squamous cell carcinoma with human papillomavirus-16 induces proliferative and morphological changes in vitro. AB - BACKGROUND: Human papillomavirus has been implicated in virtually all cervical cancers and is believed to be the primary etiological factor that transforms cervical epithelia. The presence of HPV in oral cancers suggests that HPV may play a similar role in transforming the oral epithelia. The prevalence of HPV in oral cancers is highly variable, however, presenting problematic issues regarding the etiology of oral cancers, which must be investigated more thoroughly. Past analyses of HPV in cancers of the oral cavity have largely been confined to retrospective studies of cancer patients. The purpose of this study was to examine the potential for HPV16 infection to alter the proliferative phenotype of oral squamous cell carcinoma in vitro. RESULTS: This study found that the oral squamous cell carcinoma cell line, CAL27, transfected with HPV16, exhibited significantly increased proliferation, compared with non-transfected CAL27. The increased proliferation was observed under low density conditions, even in the absence of serum. Moreover, these effects were specific to proliferation, adhesion, and morphology, while cell viability was not affected. CONCLUSION: This study represents one of the first investigations of the effects of HPV16 infection on the proliferation, adhesion, and morphology of an oral squamous cell carcinoma cell line in vitro. The finding that HPV16 has the ability to measurably alter adhesion and proliferative potential is significant, indicating that HPV may have multiple influences on precancerous and cancerous lesions and should be explored as a risk factor and mediator of cancer phenotypes. These measurements and observations will be of benefit to researchers interested in elucidating the mechanisms of oral cancer transformation and the factors governing carcinogenesis and progression. PMID- 16716228 TI - Phylogeny based discovery of regulatory elements. AB - BACKGROUND: Algorithms that locate evolutionarily conserved sequences have become powerful tools for finding functional DNA elements, including transcription factor binding sites; however, most methods do not take advantage of an explicit model for the constrained evolution of functional DNA sequences. RESULTS: We developed a probabilistic framework that combines an HKY85 model, which assigns probabilities to different base substitutions between species, and weight matrix models of transcription factor binding sites, which describe the probabilities of observing particular nucleotides at specific positions in the binding site. The method incorporates the phylogenies of the species under consideration and takes into account the position specific variation of transcription factor binding sites. Using our framework we assessed the suitability of alignments of genomic sequences from commonly used species as substrates for comparative genomic approaches to regulatory motif finding. We then applied this technique to Saccharomyces cerevisiae and related species by examining all possible six base pair DNA sequences (hexamers) and identifying sequences that are conserved in a significant number of promoters. By combining similar conserved hexamers we reconstructed known cis-regulatory motifs and made predictions of previously unidentified motifs. We tested one prediction experimentally, finding it to be a regulatory element involved in the transcriptional response to glucose. CONCLUSION: The experimental validation of a regulatory element prediction missed by other large-scale motif finding studies demonstrates that our approach is a useful addition to the current suite of tools for finding regulatory motifs. PMID- 16716229 TI - Novel insights into the aetiology and pathophysiology of increased airway inflammation during COPD exacerbations. AB - Airway inflammation increases during acute exacerbations of COPD. Extrinsic factors, such as airway infections, increased air pollution, and intrinsic factors, such as increased oxidative stress and altered immunity may contribute to this increase. The evidence for this and the potential mechanisms by which various aetiological agents increase inflammation during COPD exacerbations is reviewed. The pathophysiologic consequences of increased airway inflammation during COPD exacerbations are also discussed. This review aims to establish a cause and effect relationship between etiological factors of increased airway inflammation and COPD exacerbations based on recently published data. Although it can be speculated that reducing inflammation may prevent and/or treat COPD exacerbations, the existing anti-inflammatory treatments are modestly effective. PMID- 16716231 TI - Stigma in response to mental disorders: a comparison of Australia and Japan. AB - BACKGROUND: There are few national or cross-cultural studies of the stigma associated with mental disorders. Australia and Japan have different systems of psychiatric health care, and distinct differences in cultural values, but enjoy similar standards of living. This study seeks to compare the nature and extent of stigma among the public in the two countries. METHODS: A household survey of the public was conducted in each country using similar methodologies. The Australian study comprised a national survey of 3998 adults aged over 18 years. The Japanese survey involved 2000 adults aged 20 to 69 from 25 regional sites distributed across the country. Interviewees reported their personal attitudes (personal stigma, social distance) and perceptions of the attitudes of others (perceived stigma, perceived discrimination) in the community with respect to four case vignettes. These vignettes described a person with: depression; depression with suicidal ideation; early schizophrenia; and chronic schizophrenia. RESULTS: Personal stigma and social distance were typically greater among the Japanese than the Australian public whereas the reverse was true with respect to the perception of the attitudes and discriminatory behaviour of others. In both countries, personal stigma was significantly greater than perceived stigma. The public in both countries showed evidence of greater social distance, greater personal stigma and greater perceived stigma for schizophrenia (particularly in its chronic form) than for depression. There was little evidence of a difference in stigma for depression with and without suicide for either country. However, social distance was greater for chronic compared to early schizophrenia for the Australian public. CONCLUSION: Stigmatising attitudes were common in both countries, but negative attitudes were greater among the Japanese than the Australian public. The results suggest that there is a need to implement national public awareness interventions tailored to the needs of each country. The current results provide a baseline for future tracking of national stigma levels in each country. PMID- 16716230 TI - Avoidable mortality across Canada from 1975 to 1999. AB - BACKGROUND: The concept of 'avoidable' mortality (AM) has been proposed as a performance measure of health care systems. In this study we examined mortality in five geographic regions of Canada from 1975 to 1999 for previously defined avoidable disease groups that are amenable to medical care and public health. These trends were compared to mortality from other causes. METHODS: National and regional age-standardized mortality rates for ages less than 65 years were estimated for avoidable and other causes of death for consecutive periods (1975 1979, 1980-1985, 1985-1989, 1990-1994, and 1995-1999). The proportion of all cause mortality attributable to avoidable causes was also determined. RESULTS: From 1975-1979 to 1995-1999, the AM decrease (46.9%) was more pronounced compared to mortality from other causes (24.9%). There were persistent regional AM differences, with consistently lower AM in Ontario and British Columbia compared to the Atlantic, Quebec, and Prairies regions. This trend was not apparent when mortality from other causes was examined. Injuries, ischaemic heart disease, and lung cancer strongly influenced the overall AM trends. CONCLUSION: The regional differences in mortality for ages less than 65 years was attributable to causes of death amenable to medical care and public health, especially from causes responsive to public health. PMID- 16716232 TI - Topology and weights in a protein domain interaction network--a novel way to predict protein interactions. AB - BACKGROUND: While the analysis of unweighted biological webs as diverse as genetic, protein and metabolic networks allowed spectacular insights in the inner workings of a cell, biological networks are not only determined by their static grid of links. In fact, we expect that the heterogeneity in the utilization of connections has a major impact on the organization of cellular activities as well. RESULTS: We consider a web of interactions between protein domains of the Protein Family database (PFAM), which are weighted by a probability score. We apply metrics that combine the static layout and the weights of the underlying interactions. We observe that unweighted measures as well as their weighted counterparts largely share the same trends in the underlying domain interaction network. However, we only find weak signals that weights and the static grid of interactions are connected entities. Therefore assuming that a protein interaction is governed by a single domain interaction, we observe strong and significant correlations of the highest scoring domain interaction and the confidence of protein interactions in the underlying interactions of yeast and fly. Modeling an interaction between proteins if we find a high scoring protein domain interaction we obtain 1, 428 protein interactions among 361 proteins in the human malaria parasite Plasmodium falciparum. Assessing their quality by a logistic regression method we observe that increasing confidence of predicted interactions is accompanied by high scoring domain interactions and elevated levels of functional similarity and evolutionary conservation. CONCLUSION: Our results indicate that probability scores are randomly distributed, allowing to treat static grid and weights of domain interactions as separate entities. In particular, these finding confirms earlier observations that a protein interaction is a matter of a single interaction event on domain level. As an immediate application, we show a simple way to predict potential protein interactions by utilizing expectation scores of single domain interactions. PMID- 16716233 TI - Variation in recruitment across sites in a consent-based clinical data registry: lessons from the Canadian Stroke Network. AB - BACKGROUND: In earlier work, we found important selection biases when we tried to obtain consent for participation in a national stroke registry. Recognizing that not all registries will be exempt from requiring consent for participation, we examine here in greater depth the reasons for the poor accrual of patients from a systems perspective with a view to obtaining as representative sample as possible. METHODS: We determined the percent of eligible patients who were approached to participate and, among those approached, the percent who actually consented to participate. In addition we examined the reasons why people were not approached or did not consent and the variation across sites in the percent of patients approached and consented. We also considered site variation in restrictions on the accrual and data collection process imposed by either the local research ethics board or the hospital. RESULTS: Seventy percent of stroke patients were approached, with wide variations in approach rates across sites (from: 41% to 86%), and considerable inter-site variation in hospital policies governing patient accrual. Chief reasons for not approaching were discharge or death before being approached for consent. Seventeen percent of those approached refused to participate (range: 5% to 75%). Finally, 11% of those approached did not participate due to language or communication difficulties. CONCLUSION: We found wide variation in approach and agree rates across sites that were accounted for, in part, by different approaches to accrual and idiosyncratic policies of the hospitals. This wide variation in approach and agree rates raises important challenges for research ethics boards and data protection authorities in determining when to waive consent requirements, when to press for increased quality control, when to permit local adaptation of the consent process, and when to permit alternatives to individual express consent. We offer several suggestions for those registries that require consent for participation. PMID- 16716234 TI - Structural effects and potential changes in growth factor signalling in penis projecting autonomic neurons after axotomy. AB - BACKGROUND: The responses of adult parasympathetic ganglion neurons to injury and the neurotrophic mechanisms underlying their axonal regeneration are poorly understood. This is especially relevant to penis-projecting parasympathetic neurons, which are vulnerable to injury during pelvic surgery such as prostatectomy. We investigated the changes in pelvic ganglia of adult male rats in the first week after unilateral cavernous (penile) nerve axotomy (cut or crush lesions). In some experiments FluoroGold was injected into the penis seven days prior to injury to allow later identification of penis-projecting neurons. Neurturin and glial cell line-derived neurotrophic factor (GDNF) are neurotrophic factors for penile parasympathetic neurons, so we also examined expression of relevant receptors, GFRalpha1 and GFRalpha2, in injured pelvic ganglion neurons. RESULTS: Axotomy caused prolific growth of axon collaterals (sprouting) in pelvic ganglia ipsilateral to the injury. These collaterals were most prevalent in the region near the exit of the penile nerve. This region contained the majority of FluoroGold-labelled neurons. Many sprouting fibres formed close associations with sympathetic and parasympathetic pelvic neurons, including many FluoroGold neurons. However immunoreactivity for synaptic proteins could not be demonstrated in these collaterals. Preganglionic terminals showed a marked loss of synaptic proteins, suggesting a retrograde effect of the injury beyond the injured neurons. GFRalpha2 immunofluorescence intensity was decreased in the cytoplasm of parasympathetic neurons, but GFRalpha1 immunofluorescence was unaffected in these neurons. CONCLUSION: These studies show that there are profound changes within the pelvic ganglion after penile nerve injury. Sprouting of injured postganglionic axons occurs concurrently with structural or chemical changes in preganglionic terminals. New growth of postganglionic axon collaterals within the ganglion raises the possibility of the formation of aberrant synaptic connections between injured and un-injured ganglion neurons. Together these changes demonstrate a broader effect on the pelvic autonomic circuitry than simply loss of neuroeffector connections. These structural changes are accompanied by potential changes in neurotrophic factor signalling due to altered expression of receptors for members of the GDNF family. Together our results advance understanding of the responses of pelvic autonomic nerve circuits to injury and may assist in designing strategies for promoting regeneration. PMID- 16716235 TI - Racial and ethnic disparities in the control of cardiovascular disease risk factors in Southwest American veterans with type 2 diabetes: the Diabetes Outcomes in Veterans Study. AB - BACKGROUND: Racial/ethnic disparities in cardiovascular disease complications have been observed in diabetic patients. We examined the association between race/ethnicity and cardiovascular disease risk factor control in a large cohort of insulin-treated veterans with type 2 diabetes. METHODS: We conducted a cross sectional observational study at 3 Veterans Affairs Medical Centers in the American Southwest. Using electronic pharmacy databases, we randomly selected 338 veterans with insulin-treated type 2 diabetes. We collected medical record and patient survey data on diabetes control and management, cardiovascular disease risk factors, comorbidity, demographics, socioeconomic factors, psychological status, and health behaviors. We used analysis of variance and multivariate linear regression to determine the effect of race/ethnicity on glycemic control, insulin treatment intensity, lipid levels, and blood pressure control. RESULTS: The study cohort was comprised of 72 (21.3%) Hispanic subjects (H), 35 (10.4%) African Americans (AA), and 226 (67%) non-Hispanic whites (NHW). The mean (SD) hemoglobin A1c differed significantly by race/ethnicity: NHW 7.86 (1.4)%, H 8.16 (1.6)%, AA 8.84 (2.9)%, p = 0.05. The multivariate-adjusted A1c was significantly higher for AA (+0.93%, p = 0.002) compared to NHW. Insulin doses (unit/day) also differed significantly: NHW 70.6 (48.8), H 58.4 (32.6), and AA 53.1 (36.2), p < 0.01. Multivariate-adjusted insulin doses were significantly lower for AA (-17.8 units/day, p = 0.01) and H (-10.5 units/day, p = 0.04) compared to NHW. Decrements in insulin doses were even greater among minority patients with poorly controlled diabetes (A1c > or = 8%). The disparities in glycemic control and insulin treatment intensity could not be explained by differences in age, body mass index, oral hypoglycemic medications, socioeconomic barriers, attitudes about diabetes care, diabetes knowledge, depression, cognitive dysfunction, or social support. We found no significant racial/ethnic differences in lipid or blood pressure control. CONCLUSION: In our cohort, insulin-treated minority veterans, particularly AA, had poorer glycemic control and received lower doses of insulin than NHW. However, we found no differences for control of other cardiovascular disease risk factors. The diabetes treatment disparity could be due to provider behaviors and/or patient behaviors or preferences. Further research with larger sample sizes and more geographically diverse populations are needed to confirm our findings. PMID- 16716237 TI - Retroperitoneal metastatic squamous cell carcinoma of the tonsil (with elevated beta human chorionic gonadotrophin): a misdiagnosis as extra-gonadal germ cell tumour. AB - Head and neck cancers usually spread first to the regional lymph nodes but rarely may metastasize to distant sites. Metastasis to distant lymph node groups is a rare event. Furthermore, delayed multiple metastases without local recurrence is relatively uncommon. A case of retroperitoneal metastasis from a squamous cell carcinoma of the tonsil, secreting beta human chorionic gonadotrophin (beta-hCG), is reported. A 58-year-old man had undergone a tonsillectomy and chemo radiotherapy for squamous cell carcinoma of the left tonsil and 13 months later presented with non-specific abdominal pain. The serum beta-hCG levels were high and an abdominal ultrasound scan revealed hydronephrosis on the left side. A computed tomography scan demonstrated para-aortic retroperitoneal lymphadenopathy. The patient underwent an open lymph node biopsy. The initial pathological analysis was interpreted as extra-gonadal germ cell tumour and the patient received chemotherapy. A subsequent review was consistent with a metastatic squamous cell carcinoma of the tonsil, as immunohistochemical studies showed positive staining for epithelial membrane antigen and cytokeratins 5/6 but a negative reaction to placental alkaline phosphatase. Following this, the chemotherapy regimen was changed; however, a restaging scan demonstrated progression, and the patient died from aspiration pneumonia secondary to alcohol intoxication. To our knowledge, this is the first reported case of retroperitoneal metastasis from a squamous cell carcinoma of the tonsil, secreting beta-hCG and causing hydronephrosis. This case highlights the necessity of using clinical, histological, immunohistological and ultrastructural examination to establish precise diagnosis and to avoid inappropriate treatment. PMID- 16716236 TI - Genetic diversity among five T4-like bacteriophages. AB - BACKGROUND: Bacteriophages are an important repository of genetic diversity. As one of the major constituents of terrestrial biomass, they exert profound effects on the earth's ecology and microbial evolution by mediating horizontal gene transfer between bacteria and controlling their growth. Only limited genomic sequence data are currently available for phages but even this reveals an overwhelming diversity in their gene sequences and genomes. The contribution of the T4-like phages to this overall phage diversity is difficult to assess, since only a few examples of complete genome sequence exist for these phages. Our analysis of five T4-like genomes represents half of the known T4-like genomes in GenBank. RESULTS: Here, we have examined in detail the genetic diversity of the genomes of five relatives of bacteriophage T4: the Escherichia coli phages RB43, RB49 and RB69, the Aeromonas salmonicida phage 44RR2.8t (or 44RR) and the Aeromonas hydrophila phage Aeh1. Our data define a core set of conserved genes common to these genomes as well as hundreds of additional open reading frames (ORFs) that are nonconserved. Although some of these ORFs resemble known genes from bacterial hosts or other phages, most show no significant similarity to any known sequence in the databases. The five genomes analyzed here all have similarities in gene regulation to T4. Sequence motifs resembling T4 early and late consensus promoters were observed in all five genomes. In contrast, only two of these genomes, RB69 and 44RR, showed similarities to T4 middle-mode promoter sequences and to the T4 motA gene product required for their recognition. In addition, we observed that each phage differed in the number and assortment of putative genes encoding host-like metabolic enzymes, tRNA species, and homing endonucleases. CONCLUSION: Our observations suggest that evolution of the T4-like phages has drawn on a highly diverged pool of genes in the microbial world. The T4-like phages harbour a wealth of genetic material that has not been identified previously. The mechanisms by which these genes may have arisen may differ from those previously proposed for the evolution of other bacteriophage genomes. PMID- 16716238 TI - Reinke's oedema, hormones and hormone replacement therapy. AB - OBJECTIVE: To study the implications of hormone replacement therapy (HRT) treatment in the pathogenesis of Reinke's oedema (via a possible pseudo hypothyroidism effect), and also to study why the disease affects a predominantly post-menopausal female population. DESIGN: Prospective case series study. SETTING: Two teaching hospitals and two district general hospitals in Scotland, UK. SUBJECTS: Thirty-three patients diagnosed with Reinke's oedema who presented in the out-patient department before or after treatment. RESULTS: Thyroid function tests were normal in all but two cases. Only three patients were receiving HRT treatment. CONCLUSIONS: The study produced no evidence to support a relationship between HRT treatment and the pathogenesis of Reinke's oedema; this supports previous studies which concluded that thyroid function was not related to the development of Reinke's oedema. Some new ideas regarding hormonal factors in Reinke's oedema are discussed. PMID- 16716239 TI - A comparative study of patient satisfaction with day case and in-patient major ear surgery. AB - INTRODUCTION: Major ear surgery can be safely performed on a day case basis (i.e. six hour stay). This study aimed to ascertain whether patients had the same level of satisfaction and speed of recovery following major ear surgery when it was performed as a day case compared with performance as an in-patient procedure. METHOD: A cross-sectional survey, by postal questionnaire, of patient satisfaction with day case and in-patient major ear surgery was carried out with 158 patients. Comparisons were made between the responses of the two groups. RESULTS: The response rate was 71 per cent. Patients returned to work significantly sooner following day surgery (p < 0.025) but felt their operation to be of significantly more benefit following in-patient surgery (p < 0.05). CONCLUSIONS: Patient satisfaction following day case major ear surgery is as good as that following in-patient surgery. This has supported the expansion of this service in our unit. PMID- 16716240 TI - Prognostic relevance of anamnestic and diagnostic parameters in low-frequency hearing impairment. AB - Low-frequency hearing impairment (LFHI) is mainly attributed to endolymphatic hydrops and has a great variety of possible outcomes. At present, no conservative therapeutic regimen has proven to be 'gold-standard', and information about the prognostic indicators of LFHI is scarce. In a retrospective investigation, we evaluated the records of 90 patients who had been treated with infusions improving blood perfusion. In patients lacking complete remission, dehydration infusion therapy was added. We also undertook audiometric follow up. We calculated the outcomes after infusion therapy, dehydration therapy and after long-time hearing follow up, and we determined the prognostic relevance of several parameters of anamnesis and clinical examination to outcomes, for both therapeutic interventions and long-time hearing. The prognosis of LFHI was significantly correlated to certain anamnestic and clinical parameters; a short duration of the disease, lack of vertigo and female gender implied a better outcome. The pretherapeutic hearing threshold was an important prognostic factor; the outcome was significantly worse in patients with distinct hearing impairment in low or high frequencies, compared with that in patients with little hearing loss. Whereas vertigo was a negative prognostic factor, the results of quantitative vestibular testing were irrelevant to the outcome. The glycerol test failed to predict the effectiveness of dehydration therapy and lacked any value in predicting prognosis. These results allow the clinician to focus the anamnesis and diagnostic examination on prognostically relevant parameters, thus enabling a better estimation of the long-term disease course and improved counselling of patients. Furthermore, these results help to distinguish valuable from irrelevant diagnostic procedures. PMID- 16716241 TI - Actinomycosis of the tongue: a diagnostic dilemma. AB - Actinomycotic infections of the cervicofacial region are not uncommon. However, actinomycosis of the tongue is rare. A mass that may mimic both benign and malignant neoplasms can be seen at clinical presentation and may mislead the diagnosis. We report a patient who presented with a tumour-like tongue mass causing speech disturbance, diagnosed as actinomycosis. PMID- 16716242 TI - Evaluation of a new diagnostic modality for distal assessment of advanced, obstructing hypopharyngeal cancer. AB - INTRODUCTION: Pre-operative endoscopic assessment of the distal extension of hypopharyngeal cancer is essential for proper surgical extirpation. This assessment is frequently not feasible in advanced, obstructing tumours. AIMS: To study the role of a proposed new diagnostic technique: intra-operative open oesophagoscopy, in distal assessment of advanced hypopharyngeal cancer. MATERIALS AND METHODS: A clinicopathological study, including 35 consecutive patients with obstructing hypopharyngeal cancer. RESULTS: Intra-operative open oesophagoscopy revealed inferior submucosal tumour extension in 19 out of 22 cases proven histopathologically, with a sensitivity, specificity and accuracy of 86, 100 and 91 per cent, respectively. Oesophageal skip lesions were detected in two cases. Intra-operative open oesophagoscopy findings surpassed data obtained from pre operative radiological investigations and influenced the extent of resection performed. Accordingly, 19 patients had a total laryngopharyngectomy for local disease control, while 16 patients needed an additional total oesophagectomy. Histopathologically negative inferior resection margins were obtained in all cases. CONCLUSIONS: Intra-operative open oesophagoscopy was found to be a reliable diagnostic modality for distal assessment of obstructing hypopharyngeal cancer in cases in which pre-operative distal endoscopic examination was not feasible. PMID- 16716243 TI - Bicycle helmets: lack of efficacy against brain injury. AB - This paper is a rebuttal of the criticism by Hagel and Pless of my 2005 article in which I dispute the conclusion of a Cochrane Collaboration review that all types of standard bicycle helmet protect against injury to the brain. The main ground of rebuttal is that my critics take the relevant efficacy of helmets as given and argue from there. PMID- 16716244 TI - Effect of suramin on trypomastigote forms of Trypanosoma cruzi: changes on cell motility and on the ultrastructure of the flagellum-cell body attachment region. AB - Suramin has been previously reported to inhibit distinct cell enzymes and to affect the synthesis and distribution of cytoskeleton proteins. Our study indicates that prolonged incubation of Trypanosoma cruzi infected-LLC-MK2 cells in the presence of 500 microM suramin during the intracellular development of the parasite caused morphological changes on trypomastigote forms characterized by a partial or complete detachment of the flagellum from the cell body, besides an accentuated decrease on parasite motility. Immunofluorescence analysis of the region of adhesion between the cell body and the flagellum on trypomastigotes obtained from suramin-treated host cells after the completion of cell cycle did not show any difference in the localization of FAZ antigens recognized by 4D9 and L3B2 monoclonal antibodies despite the presence of a detached flagellum. On the other hand, suramin caused a significant increase on the phenotypic expression of FRA antigen, which was observed throughout the surface of trypomastigotes. Cytochemical localization of cationized ferritin in trypomastigotes obtained from suramin-treated host cells showed that anionic particles gained access to the space between the cell and flagellar membranes, as well as to the flagellar pocket, indicating an alteration on extracellular components of the region of adhesion between the cell body and the flagellum. PMID- 16716245 TI - [Long term results of treatment by simple surgical closure of perforated gastroduodenal ulcer followed by eradication of Helicobacter pylori]. PMID- 16716246 TI - Influences of animal mucins on lysozyme activity in solution and on hydroxyapatite surfaces. AB - The purpose of this study was to investigate the influence of animal mucins on lysozyme activity in solution and on the surface of hydroxyapatite (HA) beads. The effects of animal mucins on lysozyme activity in solution were examined by incubating porcine gastric mucin (PGM) or bovine submaxillary mucin (BSM) with hen egg-white lysozyme (HEWL) or salivary samples. HA-immobilised animal mucins or lysozyme were used to determine the influence of animal mucins on lysozyme activity on HA surfaces. Lysozyme activity was determined by turbidity measurement of a Micrococcus lysodeikticus substrate suspension. Protein concentration was determined by ninhydrin assay. PGM inhibited the activity of HEWL and salivary lysozyme in solution. The amount of inhibition was dependent on mucin concentration, incubation time and temperature, and the structural integrity of the mucin. The inhibition of salivary lysozyme activity by PGM was greater in submandibular/sublingual saliva than in parotid saliva. The inhibition of lysozyme activity by PGM was markedly dependent on pH. However, BSM did not inhibit the in-solution lysozyme activities of HEWL and clarified saliva. Both PGM and BSM bound to HA surfaces, and HA-adsorbed animal mucins increased the subsequent adsorption of lysozyme. When HA beads were exposed to a mixture of HEWL and PGM or BSM, lysozyme activity on the HA surfaces was significantly increased. The results suggest that animal mucins affect lysozyme activity, and the effects are different on HA surfaces compared with in solution. Further research is needed to determine the effect of animal mucins on lysozyme activity in vivo. PMID- 16716247 TI - Mitochondrial membrane permeability transition and cell death. AB - Mitochondria are important organelles for energy production, Ca2+ homeostasis, and cell death. In recent years, the role of the mitochondria in both apoptotic and necrotic cell death has received much attention. In apoptotic and necrotic death, an increase of mitochondrial membrane permeability is considered to be one of the key events, although the detailed mechanism remains to be elucidated. The mitochondrial membrane permeability transition (MPT) is a Ca2+-dependent increase in the permeability of the mitochondrial membrane that leads to loss of Deltapsi, mitochondrial swelling, and rupture of the outer mitochondrial membrane. The MPT is thought to occur after the opening of a channel, which is termed the permeability transition pore (PTP) and putatively consists of the voltage dependent anion channel (VDAC), the adenine nucleotide translocator (ANT), cyclophilin D (Cyp D: a mitochondrial peptidyl prolyl-cis, trans-isomerase), and other molecule(s). Our studies of mice lacking Cyp D have revealed that it is essential for occurrence of the MPT and that the Cyp D-dependent MPT regulates some forms of necrotic cell death, but not apoptotic death. We have also shown that two anti-apoptotic proteins, Bcl-2 and Bcl-x(L), block the MPT by directly inhibition of VDAC activity. Here we summarize a role of the MPT in cell death. PMID- 16716248 TI - LL-37, the only human member of the cathelicidin family of antimicrobial peptides. AB - Antimicrobial peptides and their precursor molecules form a central part of human and mammalian innate immunity. The underlying genes have been thoroughly investigated and compared for a considerable number of species, allowing for phylogenetic characterization. On the phenotypical side, an ever-increasing number of very varied and distinctive influences of antimicrobial peptides on the innate immune system are reported. The basic biophysical understanding of mammalian antimicrobial peptides, however, is still very limited. This is especially unsatisfactory since knowledge of structural properties will greatly help in the understanding of their immunomodulatory functions. The focus of this review article will be on LL-37, the only cathelicidin-derived antimicrobial peptide found in humans. LL-37 is a 37-residue, amphipathic, helical peptide found throughout the body and has been shown to exhibit a broad spectrum of antimicrobial activity. It is expressed in epithelial cells of the testis, skin, the gastrointestinal tract, and the respiratory tract, and in leukocytes such as monocytes, neutrophils, T cells, NK cells, and B cells. It has been found to have additional defensive roles such as regulating the inflammatory response and chemo attracting cells of the adaptive immune system to wound or infection sites, binding and neutralizing LPS, and promoting re-epthelialization and wound closure. The article aims to report the known biophysical facts, with an emphasis on structural evidence, and to set them into relation with insights gained on phylogenetically related antimicrobial peptides in other species. The multitude of immuno-functional roles is only outlined. We believe that this review will aid the future work on the biophysical, biochemical and immunological investigations of this highly intriguing molecule. PMID- 16716249 TI - Electrostatic interactions of colicin E1 with the surface of Escherichia coli total lipid. AB - The surface properties of colicin E1, a 522-amino acid protein, and its interaction with monolayers of Escherichia coli (E. coli) total lipid and 1,2 Dimyristoyl-sn-Glycero-3-Phosphocholine (DOPC) were studied using the Langmuir Blodgett (LB) technique. Colicin E1 is amphiphilic, forming a protein monolayer at the air/buffer interface. The protein is thought to interact with the E. coli total lipid head groups through electrostatic interactions, followed by its insertion into the lipid monolayers. Supported lipid bilayers (SLBs) of E. coli total lipid and DOPC, deposited onto mica at the cell membrane equivalence pressure for E. coli and incubated with colicin E1, were imaged by contact mode atomic force microscopy (CM-AFM). Colicin E1 formed protein aggregates on DOPC SLBs, while E. coli total lipid SLB was deformed following its incubation with colicin E1. Corresponding lateral force images, along with electrostatic surface potentials for colicin E1 P190, imply a direct interaction of colicin E1 with lipid head groups facilitating their charge neutralization. PMID- 16716250 TI - Conditions affecting the re-alignment of the antimicrobial peptide PGLa in membranes as monitored by solid state 2H-NMR. AB - The cationic antimicrobial peptide PGLa is electrostatically attracted to bacterial membranes, binds as an amphiphilic alpha-helix, and is thus able to permeabilize the lipid bilayer. Using solid state (2)H-NMR of non-perturbing Ala d(3) labels on the peptide, we have characterized the helix alignment under a range of different conditions. Even at a very high peptide-to-lipid ratio (1:20) and in the presence of negatively charged lipids, there was no indication of a toroidal wormhole structure. Instead, PGLa re-aligns from a surface-bound S-state to an obliquely tilted T-state, which is presumably dimeric. An intermediate structure half-way between the S- and T-state was observed in fully hydrated multilamellar DMPC vesicles at 1:50, suggesting a fast exchange between the two states on the time scale of >50 kHz. We demonstrate that this equilibrium is shifted from the S- towards the T-state either upon (i) increasing the peptide concentration, (ii) adding negatively charged DMPG, or (iii) decreasing the level of hydration. The threshold concentration for re-alignment in DMPC is found to be between 1:200 and 1:100 in oriented samples at 96% humidity. In fully hydrated multilamellar DMPC vesicles, it shifts to an effective peptide-to-lipid ratio of 1:50 as some peptides are able to escape into the bulk water phase. PMID- 16716251 TI - The structure, function and regulation of the nodulin 26-like intrinsic protein family of plant aquaglyceroporins. AB - The nodulin 26-like intrinsic protein family is a group of highly conserved multifunctional major intrinsic proteins that are unique to plants, and which transport a variety of uncharged solutes ranging from water to ammonia to glycerol. Based on structure-function studies, the NIP family can be subdivided into two subgroups (I and II) based on the identity of the amino acids in the selectivity-determining filter (ar/R region) of the transport pore. Both subgroups appear to contain multifunctional transporters with low to no water permeability and the ability to flux multiple uncharged solutes of varying sizes depending upon the composition of the residues of the ar/R filter. NIPs are subject to posttranslational phosphorylation by calcium-dependent protein kinases. In the case of the family archetype, soybean nodulin 26, phosphorylation has been shown to stimulate its transport activity and to be regulated in response to developmental as well as environmental cues, including osmotic stresses. NIPs tend to be expressed at low levels in the plant compared to other MIPs, and several exhibit cell or tissue specific expression that is subject to spatial and temporal regulation during development. PMID- 16716252 TI - NMR studies of aurein 1.2 analogs. AB - Aurein 1.2 is an antimicrobial and anticancer peptide isolated from an Australian frog. To improve our understanding of the mechanism of action, two series of peptides were designed. The first series includes the N-terminal membrane anchor of bacterial glucose-specific enzyme IIA, aurein 1.2, and a newly identified aurein 1.2 analog from human LL-37 (LLAA). The order of antibacterial activity is LLAA>aurein 1.2>>the membrane anchor (inactive). The structure of LLAA in detergent micelles was determined by (1)H NMR spectroscopy, including structural refinement by natural abundance (13)C(alpha), (13)C(beta), and (15)N chemical shifts. The hydrophobic surface area of the 3D structure is related to the retention time of the peptide on a reverse-phase HPLC column. The higher activity of LLAA compared to aurein 1.2 was attributed to additional cationic residues that enhance the membrane perturbation potential. The second peptide series was created by changing the C-terminal phenylalanine (F13) of aurein 1.2 to either phenylglycine or tryptophan. A closer or further location of the aromatic rings to the peptide backbone in the mutants relative to F13 is proposed to cause a drop in activity. Phenylglycine with unique chemical shifts may be a useful NMR probe for structure-activity relationship studies of antimicrobial peptides. To facilitate potential future use for NMR studies, random-coil chemical shifts for phenylglycine (X) were measured using the synthetic peptide GGXGG. Aromatic rings of phenylalanines in all the peptides penetrated 2-5 A below the lipid head group and are essential for membrane targeting as illustrated by intermolecular peptide lipid NOE patterns. PMID- 16716253 TI - Identification of phosphoproteins associated with maintenance of transformed state in temperature-sensitive Rous sarcoma-virus infected cells by proteomic analysis. AB - To identify phosphotyrosine-containing proteins essential for maintaining the transformed state, we studied the tyrosine phosphorylation profile of temperature sensitive mutant of Rous sarcoma virus, tsNY68, infected cells (68N7). Shifting the temperature from 39 degrees C (nonpermissive) to 32 degrees C (permissive) markedly increased the expression of phosphotyrosine-containing cell membrane proteins of approximately 40kDa, as assessed by SDS-PAGE. Membrane and nuclear proteins were separated by two-dimensional gel electrophoresis and immunoblotted with anti-phosphotyrosine antibody. Proteins showing temperature-dependent changes in phosphorylation profile were subjected to in-gel digestion with trypsin and analyzed by mass spectrometry. Five proteins were identified: heterogeneous nuclear ribonucleoprotein (hnRNP) A3, hnRNP A2, annexin II, phosphoglycerate mutase 1, and triosephosphate isomerase 1. hnRNP A3 was phosphorylated at serine residues and had both serine and tyrosine phosphorylated sites. These results suggest an important complementary role for proteomics in identifying molecular abnormalities associated with tumor progression that may be attractive candidates for tumor diagnosis. PMID- 16716254 TI - Molecular genetic determinants of human brain size. AB - Cognitive skills such as tool use, syntactical languages, and self-awareness differentiate humans from other primates. The underlying basis for this cognitive difference has been widely associated with a high encephalization quotient and an anatomically distinct, exceptionally large cerebral cortex. Investigations on congenital microcephaly had revealed several genes that affect mammalian brain size when mutated. At least four of these, microcephalin (MCPH1), abnormal spindle-like microcephaly-associated (ASPM), cyclin-dependent kinase 5 regulatory associated protein 2 (CDK5RAP2), and centromere-associated protein J (CENPJ) are known to have undergone significant positive selection in the great apes and human lineages during primate evolution. MCPH1 and ASPM both have very young single nucleotide polymorphism haplotypes associated with modern humans, and these genes are presumably still evolving in Homo sapiens. Microcephalin has a role in DNA damage response and regulation of cell cycle checkpoints. The other known microcephaly-associated genes encode microtubule-associated centrosomal proteins that might regulate neural progenitor cell division and cell number. Recent reports have also unveiled a previously unknown function of ephrins and Eph in the regulation of neural progenitor cell death with a consequential effect on brain size. Understanding the mechanism for developmental control of brain organogenesis by these genes, and others such as FOXP2, shall provide fresh perspectives on the evolution of human intelligence. PMID- 16716256 TI - The critical role of caspases activation in hypoxia/reoxygenation induced apoptosis. AB - Hypoxia/reoxygenation insult can be found in many tissues, including heart, brain, and tumor. It is believed that cell death may be resulted after cells were subjected to chronic hypoxia or reoxygenation after chronic hypoxia. The molecular mechanism for reoxygenation induced cell death is so far not clear and will require further study, in particular, to be distinguished from the pathways associated only with chronic hypoxia. In this study, the cell death mechanism in human squamous carcinoma A431 cells after hypoxia/reoxygenation insult is examined. It is demonstrated that although caspase-9 and -3 were activated during both hypoxia and reoxygenation, only those caspases activated during reoxygenation were responsible for reoxygenation induced apoptosis. Activation of caspase-9 and -3 during reoxygenation is believed to be triggered by the ROS formation at the time of reoxygenation. Addition of catalase during reoxygenation was found to attenuate reoxygenation induced apoptosis and caspase activation. PMID- 16716257 TI - Novel hydroxyl radical scavenging antioxidant activity assay for water-soluble antioxidants using a modified CUPRAC method. AB - Reactive oxygen species (ROS) such as superoxide anion, hydroxyl ((*)OH), peroxyl, and alkoxyl radicals may attack biological macromolecules giving rise to oxidative stress-originated diseases. Since (*)OH is very short-lived, secondary products resulting from (*)OH attack to various probes are measured. Although the measurement of aromatic hydroxylation with HPLC/electrochemical detection is more specific than the low-yield TBARS test, it requires sophisticated instrumentation. As a more convenient and less costly alternative, we used p aminobenzoate, 2,4- and 3,5-dimethoxybenzoate probes for detecting hydroxyl radicals generated from an equivalent mixture of Fe(II)+EDTA with hydrogen peroxide. The produced hydroxyl radicals attacked both the probe and the water soluble antioxidants in 37 degrees C-incubated solutions for 2h. The CUPRAC (i.e., our original method for total antioxidant capacity assay) absorbance of the ethylacetate extract due to the reduction of Cu(II)-neocuproine reagent by the hydroxylated probe decreased in the presence of (*)OH scavengers, the difference being proportional to the scavenging ability of the tested compound. A rate constant for the reaction of the scavenger with hydroxyl radical can be deduced from the inhibition of color formation. The second-order rate constants of the scavengers were determined with competition kinetics by means of a linear plot of A(0)/A as a function of C(scavenger)/C(probe), where A(0) and A are the CUPRAC absorbances of the system in the absence and presence of scavenger, respectively, and C is the molar concentration of relevant species. The 2,4- and 3,5-dimethoxybenzoates were the best probes in terms of linearity and sensitivity. Iodide, metabisulfite, hexacyanoferrate(II), thiourea, formate, and dimethyl sulfoxide were shown by the modified CUPRAC assay to be more effective scavengers than mannitol, glucose, lysine, and simple alcohols, as in the TBARS assay. The developed method is less lengthy, more specific, and of a higher yield than the classical TBARS assay. The hydroxyl radical scavenging rate constants of ascorbic acid, formate, and hexacyanoferrate(II) that caused interference in other assays could be easily found with the proposed procedure. PMID- 16716255 TI - Anti-inflammatory effect of roasted licorice extracts on lipopolysaccharide induced inflammatory responses in murine macrophages. AB - Licorice, the roots of Glycyrrhiza inflata, is used by practitioners of alternative medicine to treat individuals with gastric or duodenal ulcers, bronchitis, cough, arthritis, adrenal insufficiency, and allergies. We investigated the anti-inflammatory properties of 4 licorice extracts: extracts of roasted licorice obtained by ethanol (rLE) or water extraction (rLW) and extracts of raw licorice obtained by ethanol (LE) or water extraction (LW). rLE demonstrated strong anti-inflammatory activity through its ability to reduce nitric oxide and prostaglandin E(2) production in the LPS-stimulated mouse macrophage cell, RAW264.7. It also inhibited the production of pro-inflammatory cytokines and CD14 expression on the LPS-stimulated RAW264.7 cells. Further study indicated that LPS-induced degradation and phosphorylation of Ikappa-Balpha, along with DNA-binding of NF-kappaB, was significantly inhibited by rLE exposure in RAW264.7 cells. In the murine model, we found that in vivo exposure to rLE induced an increase in the survival rate, reduced plasma levels of TNF-alpha and IL-6, and increased IL-10 production in LPS-treated mice. Collectively, these data suggest that the use of rLE may be a useful therapeutic approach to various inflammatory diseases. PMID- 16716258 TI - TIMP-2 promotes cell spreading and adhesion via upregulation of Rap1 signaling. AB - We previously demonstrated that TIMP-2 treatment of human microvascular endothelial cells (hMVECs) activates Rap1 via the pathway of paxillin-Crk-C3G. Here, we show that TIMP-2 overexpression in hMVECs by adenoviral infection enhances Rap1 expression, leading to further increase in Rap1-GTP. TIMP-2 expression, previously reported to inhibit cell migration, also leads to cell spreading accompanied with increased cell adhesion. HMVECs stably expressing Rap1 display a similar phenotype as hMVECs-TIMP-2, whereas the expression of inactive Rap1 mutant, Rap1(38N), leads to elongated appearance with greatly reduced cell adhesion. Furthermore, the phenotype of hMVECs-Rap1(38N) was not reversed by TIMP 2 overexpression. TIMP-2 greatly promotes the association of Rap1 with actin. Therefore, these findings suggest that TIMP-2 mediated alteration in cell morphology requires Rap1, TIMP-2 may recruit Rap1 to sites of actin cytoskeleton remodeling necessary for cell spreading, and enhanced cell adhesion by TIMP-2 expression may hinder cell migration. PMID- 16716259 TI - Elevated extracellular calcium stimulates secretion of bone morphogenetic protein 2 by a macrophage cell line. AB - It has been suggested that macrophages and multinucleated giant cells are responsible for phagocytosis of resorbable calcium phosphate (CaP) compounds implanted in bone defects. However, function of macrophages around the CaP, if continuously exposed to various concentration of extracellular calcium ions ([Ca(2+)](o)), is still unknown. The present study showed that when resorbable octacalcium phosphate was implanted in mouse calvaria, macrophage-like cells were observed around the implant during bone formation. Then, experiments were designed to investigate whether secretion of bone morphogenetic protein 2 (BMP-2) is enhanced by [Ca(2+)](o) in a macrophage cell line (J774A.1) in vitro. The mRNA expression and the secretion of BMP-2 in J774A.1 cells were significantly increased when incubated in the medium with [Ca(2+)](o) up to 14mM. The promotion of mRNA expression was maintained even when incubated with a small amount of minute CaP crystals. The present results suggest that [Ca(2+)](o) above physiological concentration may stimulate macrophages to induce osteogenic cytokine, such as BMP-2, for bone formation by osteoblast. PMID- 16716260 TI - Murine leukemia virus particles activate Rac1 in HeLa cells. AB - A number of viruses, when they bind to cells, activate intracellular signals that facilitate post-binding steps of infection. To determine if retroviruses activate intracellular signaling, we transduced HeLa cells with amphotropic retroviruses produced by TelCeB6 cells and examined cell lysates for activated Rac1. We found that retroviruses activate Rac1. Rac1 activation was blocked when cells were depleted of cholesterol, cultured in suspension, or incubated with an anti beta(1) integrin antibody, and when viruses were treated with heparinase III. Retrovirus activation of Rac1 did not require the amphotropic envelope protein. Gene transfer was reduced 2.4-fold when viruses were treated with heparinase III, but did not change when cells were transduced in the presence of function blocking anti-beta(1) integrin antibodies. The implications of these findings with respect to retrovirus-cell interactions are discussed. PMID- 16716261 TI - Purification and identification of a BMP-like factor from bovine serum. AB - Myogenic differentiation is suppressed in vitro by unknown factors present in fetal bovine serum (FBS). We found that specific inhibitors of bone morphogenetic proteins (BMPs) stimulated myogenic differentiation even in the presence of 20% FBS, which in turn activated specific BMP signaling. Moreover, these specific BMP inhibitors blocked maturation of osteoblastic cells induced by FBS, indicating that BMP-like factor(s) in serum regulate both myogenic and osteoblastic differentiation. The factor identified had an apparent molecular weight (Mw) of over 100kDa on a Superdex 200 column for molecular sieving HPLC, but an apparent Mw of 33kDa on SDS-PAGE under non-reducing conditions. Analysis of a purified preparation from FBS (5L) by liquid chromatography-tandem mass spectrometry revealed the presence of an amino acid sequence conserved between mature human and murine BMP-4. This is the first study to show that BMP-4 is present in FBS as a large complex. PMID- 16716262 TI - Exo-Dye-based assay for rapid, inexpensive, and sensitive detection of DNA binding proteins. AB - We reported herein a rapid, inexpensive, and sensitive technique for detecting sequence-specific DNA-binding proteins. In this technique, the common exonuclease III (ExoIII) footprinting assay is coupled with simple SYBR Green I staining for monitoring the activities of DNA-binding proteins. We named this technique as ExoIII-Dye-based assay. In this assay, a duplex probe was designed to detect DNA binding protein. One side of the probe contains one protein-binding site, and another side of it contains five protruding bases at 3' end for protection from ExoIII digestion. If a target protein is present, it will bind to binding sites of probe and produce a physical hindrance to ExoIII, which protects the duplex probe from digestion of ExoIII. SYBR Green I will bind to probe, which results in high fluorescence intensity. On the contrary, in the absence of the target protein, the naked duplex probe will be degraded by ExoIII. SYBR Green I will be released, which results in a low fluorescence intensity. In this study, we employed this technique to successfully detect transcription factor NF-kappaB in crude cell extracts. Moreover, it could also be used to evaluate the binding affinity of NF-kappaB. This technique has therefore wide potential application in research, medical diagnosis, and drug discovery. PMID- 16716263 TI - IGF-I and IGF-II stimulate directed cell migration of bone-marrow-derived human mesenchymal progenitor cells. AB - Insulin-like growth factors (IGFs) are known to be key regulators of bone growth, remodeling, and repair. Since all these processes depend on the recruitment of cells with the potential to be committed to the osteoblastic lineage, we studied possible effects of IGF-I and -II on migration of human mesenchymal progenitor cells (MPC) using a modified Boyden chamber assay. The results were compared to those of primary osteoblasts and in vitro-osteogenic-differentiated MPC. IGF-I and -II stimulated cell migration of all these cell populations in a dose dependent manner from 1 to 100ng/mL. The maximal chemotactic index (CI) was 4-5 for MPC and primary osteoblasts and about 3 for in vitro-differentiated MPC. Checkerboard analysis revealed that IGFs stimulated true directed cell migration (chemotaxis) and not simply chemokinesis. Addition of an antibody against the type I IGF receptor (alphaIR3) completely abolished (MPC) or markedly reduced (primary osteoblasts) the chemotactic effects of each of the IGFs. IGFBP-3 itself had no direct effect, while IGFBP-5 stimulated MPC migration at concentrations of 80 and 160ng/mL. Parallel application of IGFBP-3 had borderline inhibitory effects while the addition of 40ng/mL of IGFBP-5 enhanced the chemotactic effect of IGF-I on MPC. In conclusion, our results show that IGF-I and -II are chemotactic factors for MPC and indicate that IGFBP-5 both modulates the IGF-I effect and directly stimulates migration of human mesenchymal progenitor cells. PMID- 16716265 TI - Toxin insights into nicotinic acetylcholine receptors. AB - Venomous species have evolved cocktails of bioactive peptides to facilitate prey capture. Given their often exquisite potency and target selectivity, venom peptides provide unique biochemical tools for probing the function of membrane proteins at the molecular level. In the field of the nicotinic acetylcholine receptors (nAChRs), the subtype specific snake alpha-neurotoxins and cone snail alpha-conotoxins have been widely used to probe receptor structure and function in native tissues and recombinant systems. However, only recently has it been possible to generate an accurate molecular view of these nAChR-toxin interactions. Crystal structures of AChBP, a homologue of the nAChR ligand binding domain, have now been solved in complex with alpha-cobratoxin, alpha conotoxin PnIA and alpha-conotoxin ImI. The orientation of all three toxins in the ACh binding site confirms many of the predictions obtained from mutagenesis and docking simulations on homology models of mammalian nAChR. The precise understanding of the molecular determinants of these complexes is expected to contribute to the development of more selective nAChR modulators. In this commentary, we review the structural data on nAChR-toxin interactions and discuss their implications for the design of novel ligands acting at the nAChR. PMID- 16716264 TI - Liver PPARalpha and UCP2 are involved in the regulation of obesity and lipid metabolism by swim training in genetically obese db/db mice. AB - Swim training for 6 weeks significantly decreased body weight gain, adipose tissue mass, and adipocyte size in both sexes of genetically obese db/db mice compared with their respective sedentary controls. Swim training also caused significant decreases in serum levels of free fatty acids, triglycerides, and total cholesterol in both sexes of obese mice. Concomitantly, hepatic mRNA levels of peroxisome proliferator-activated receptor alpha (PPARalpha) target enzymes responsible for mitochondrial and peroxisomal fatty acid beta-oxidation were significantly increased by swim training. Moreover, mRNA levels of uncoupling protein 2 (UCP2) in liver were also markedly increased by swim training. In conclusion, these results suggest that swim training-induced transcriptional activation of hepatic PPARalpha target enzymes and UCP2 may effectively prevent body weight gain, adiposity, and lipid disorders caused by leptin receptor deficiency in both sexes of mice. PMID- 16716266 TI - Effect of opioid receptor ligands injected into the rostral lateral hypothalamus on c-fos and feeding behavior. AB - The lateral hypothalamic area (LHa) is an important brain site for the regulation of food intake. Central injection of opioids increases food intake, and the LHa contains mu and kappa opioid receptors, both of which are involved in feeding behavior. It is unclear whether opioids impact feeding when injected directly into the rostral portion of the LHa (rLHa) in rats. We performed a series of studies in which free-feeding rLHa-cannulated rats were injected with opioid agonists (DAMGO, morphine, dynorphin, U-50488H) followed by the measurement of food intake at 1, 2, and 4 h postinjection. To determine whether opioid receptor ligands administered into the rLHa affect neuronal activation in this brain site, we studied cFos immunoreactivity (cFos IR) in response to rLHa stimulation with naltrexone. We found that the only compound that stimulated feeding behavior was morphine. The other agonists had no effect on food consumption. Naltrexone injection into the rLHa increased neural activation in the LHa, indicating the presence of functional opioid receptors in this region. These data suggest that although neuronal activity is affected by opioid agents acting in the rLHa, administration of selective mu and kappa opioid ligands in this subdivision of the LHa does not have a reliable effect on feeding behavior. PMID- 16716267 TI - Depression of NMDA-receptor-mediated segmental transmission by ketamine and ketoprofen, but not L-NAME, on the in vitro neonatal rat spinal cord preparation. AB - Activation of spinal N-methyl-D-aspartate (NMDA) receptors and then the nitric oxide and the arachidonic acid pathways is important in pain transmission. This study assessed the effects of the NMDA receptor channel blocker ketamine, the nitric oxide synthase inhibitor L-NAME, and the cyclooxygenase inhibitor ketoprofen in nociceptive transmission using an in vitro neonatal rat spinal cord preparation. Supramaximal electrical stimulation of the dorsal root evoked the A fibre- and C-fibre-mediated high intensity excitatory postsynaptic potential (EPSP) in the ipsilateral ventral root. Low intensity stimulation evoked the A fibre-mediated monosynaptic compound action potential (MSR) superimposed on the low intensity EPSP. Both the low intensity EPSP and the high intensity EPSP contain NMDA-receptor-mediated components. Only ketamine and ketoprofen depressed the synaptic responses. Ketamine depressed all three spinal reflexes with IC(50) values (with 95% CI) of 10.80 microM (5.97 to 19.54 microM) for the MSR, 8.29 microM (4.53 to 14.17 microM) for the low intensity EPSP, and 5.35 microM (3.05 to 9.40 microM) for the high intensity EPSP. Ketoprofen depressed the low intensity EPSP and the high intensity EPSP only; IC(50) values (with 95% CI) were 354.5 microM (217.5 to 576.8 microM) and 302.7 microM (174.0 to 526.7 microM), respectively. Reflexes recovered after drug washout. These data demonstrated that ketamine and ketoprofen, but not L-NAME, depressed NMDA-mediated nociceptive transmission in spinal cord preparations from neonatal rats. PMID- 16716268 TI - The adult rat hippocampal slice revisited with multi-electrode arrays. AB - The multi-electrode arrays (MEA) technology for the recording of brain slices is available for more than 10 years. However, despite its relative simplicity, this recording technique is not widely used in academic or pharmaceutical research laboratories. We illustrate here that MEA provide multiple possibilities to investigate some network physiological properties as well as to evaluate the pharmacological effects of compounds. We first document that MEA allow to trigger and to record conventional FP which are inhibited by the block of action potential propagation (with 500 nM TTX). FP recorded with MEA are sensitive to ionic substitutions, to ionotropic glutamate receptor antagonists (CNQX or NBQX) and to energetic failure. Second, we illustrate that different "classical" protocols (paired-pulse, LTP, chemical LTD), revealing synaptic plasticity mechanisms, could be performed. Third, we document that MEA allow spatial and temporal discriminations for the effects of known pharmacological compounds such as competitive antagonist (gabazine, bicuculline) and allosteric modulators (steroids) of GABA(A) receptors. In conclusion, we illustrate that MEA recordings of adult rat hippocampal slices constitute a powerful and sensitive system to evaluate the effect of molecules on basic synaptic propagation/transmission and on synaptic plasticity processes. PMID- 16716269 TI - Molecular characterization and expression of a two-pore domain potassium channel in the CNS of Aplysia californica. AB - A cDNA encoding a two-pore domain potassium (K2p) channel subunit, AcK2p2, was cloned from the CNS of the marine opisthobranch Aplysia californica. This is the second K2p subunit to be identified in molluscs. Like the K2p subunit cloned previously from Aplysia, AcK2p2 appears to be more closely related to human K2p channels than to any from Drosphila melanogaster or Caenorhabditis elegans. However, the overall identity is much lower (24% with human TALK-1) and phylogenetic analysis indicates that AcK2p2 cannot be grouped into any established mammalian subclass. We analyzed the distribution of this channel by in situ hybridization in whole mount preparations of the CNS. Less than a dozen of the approximately 20,000 neurons in the CNS expressed AcK2p2 at high levels, with the consistently intense labeling seen in a single bilaterally symmetrical pair of pedal neurons. The neuron-specific expression pattern seen for this channel is consistent with data from a variety of organisms that implicate K2p channels as determinants of neuronal phenotype and function. PMID- 16716270 TI - Characteristics of IA currents in adult rabbit cerebellar Purkinje cells. AB - Classical conditioning the rabbit nictitating membrane involves changes in synaptic and intrinsic membrane properties of cerebellar Purkinje cell dendrites, and a 4-aminopyridine (4-AP)-sensitive potassium channel underlies these membrane properties. We characterized I(A) currents in adult, rabbit Purkinje cells to determine whether I(A) is the target channel involved in learning. Whole-cell recordings of Purkinje cell somas and dendrites revealed a fast activating and inactivating current with half maximal activation at -27.08 +/- 3.48 mV and 25.51 +/- 1.15 mV in somas and dendrites, respectively; half maximal inactivation at -58.91 +/- 2.34 mV and -49.90 +/- 2.58 mV; and a recovery time constant of 22.81 +/- 1.92 ms and 16.60 +/- 4.26 ms. Outside-out patch recordings from cerebellar Purkinje cell somas confirmed these 4-AP-sensitive currents with half maximal activation at -13.85 +/- 1.17 mV and half maximal inactivation at -55.07 +/- 5.54 mV. More importantly, there was an overlap of activation and incomplete inactivation at potentials from -60 to -40 mV, suggesting a "window" current that was responsible for subthreshold variations of membrane potential and might underlie conditioning-specific increases in Purkinje cell excitability. The potassium current was inhibited by 4-AP and by Heteropodatoxin, a specific blocker of Kv4.2 and Kv4.3 channels, but not by Stromatoxin, a blocker of Kv4.2 channels. Mouse monoclonal antibody labeling identified both Kv4.3 and Kv4.2 subunits in the granule cell layer but only Kv4.3 subunits in the molecular layer. This is the first demonstration of A-type currents in adult, rabbit Purkinje cells that may play a role in regulating membrane potential and firing frequency and comprise the target channel mediating conditioning-specific changes of excitability in rabbit Purkinje cell dendrites. PMID- 16716271 TI - Acceptor-dependent regioselectivity of glycosynthase reactions by Streptomyces E383A beta-glucosidase. AB - The nonnucleophilic mutant E383A beta-glucosidase from Streptomyces sp. has proven to be an efficient glycosynthase enzyme, catalyzing the condensation of alpha-glucosyl and alpha-galactosyl fluoride donors to a variety of acceptors. The enzyme has maximal activity at 45 degrees C, and a pH-dependence reflecting general base catalysis with an apparent kinetic pKa of 7.2. The regioselectivity of the new glycosidic linkage depends unexpectedly on the acceptor substrate. With aryl monosaccharide acceptors, beta-(1-->3) disaccharides are obtained in good to excellent yields, thus expanding the synthetic products available with current exo-glycosynthases. With xylopyranosyl acceptor, regioselectivity is poorer and results in the formation of a mixture of beta-(1-->3) and beta-(1-->4) linkages. In contrast, disaccharide acceptors produce exclusively beta-(1-->4) linkages. Therefore, the presence of a glycosyl unit in subsite +II redirects regioselectivity from beta-(1-->3) to beta-(1-->4). To improve operational performance, the E383A mutant was immobilized on a Ni2+-chelating Sepharose resin. Immobilization did not increase stability to pH and organic solvents, but the operational stability and storage stability were clearly enhanced for recycling and scaling-up. PMID- 16716272 TI - Isolation and structural analysis of ajugose from Vigna mungo L. AB - The hexasaccharide ajugose, alpha-D-galactopyranosyl-(1-->6)-alpha-D galactopyranosyl-(1-->6)-O-alpha-D-galactopyranosyl-(1-->6)-alpha-D galactopyranosyl-(1-->6)-alpha-D-glucopyranosyl-(1<-->2)-beta-D-fructofuranoside, generally uncommon in legumes, was detected in the seeds of Vigna mungo L. by TLC and paper chromatography. Ajugose was then isolated by silica gel chromatography and its structure was established by acid and enzymatic hydrolysis, fast atom bombardment mass spectrometry and both one- and two-dimensional 1H and 13C NMR techniques. PMID- 16716273 TI - Chemoenzymatic synthesis and application of a sialoglycopolymer with a chitosan backbone as a potent inhibitor of human influenza virus hemagglutination. AB - Sialoglycopeptide (SGP) is referred as the glycopeptide in hen's egg yolk, which has an N-linked, complex-type, disialyl biantennary oligosaccharide with an alpha (2-->6)-sialyl N-acetyllactosamine residue. The residue is known as a binding ligand of type-A human influenza virus hemagglutinin. We describe herein a simple synthesis of a sialoglycopolymer with a chitosan backbone as a potent inhibitor of human influenza virus hemagglutination that makes use of the natural source ingredient, SGP, and the transglycosylation activity of endo-beta-N acetylglucosaminidase from Mucor hiemalis (Endo-M). Its inhibitiory activity for influenza virus hemagglutination is 40 times higher than that of SGP, and its competitive inhibition is determined to be over 300 times higher than that of fetuin. These results indicate that a sialoglycopolymer having a multivalent sialo-oligosaccharide could potentially be used for the prevention of influenza virus infection. PMID- 16716274 TI - A novel one-pot 'green' synthesis of stable silver nanoparticles using soluble starch. AB - Stable silver nanoparticles have been synthesized by using soluble starch as both the reducing and stabilizing agents; this reaction was carried out in an autoclave at 15 psi, 121 degrees C for 5 min. Nanoparticles thus prepared are found to be stable in aqueous solution over a period of three months at room temperature (approximately 25 degrees C). The size of these nanoparticles was found to be in the range of 10-34 nm as analyzed using transmission electron micrographs. The X-ray diffraction analysis revealed the face-centred cubic (fcc) geometry of silver nanoparticles. Iodometric titration confirmed the entrapment of silver nanoparticles inside the helical amylose chain. These silver nanoparticles embedded in soluble starch produced a typical emission peak at 553 nm when excited at 380 nm. The use of environmentally benign and renewable materials like soluble starch offers numerous benefits of eco-friendliness and compatibility for pharmaceutical and biomedical applications. PMID- 16716275 TI - Solid-phase chemical tools for glycobiology. AB - Techniques involving solid supports have played crucial roles in the development of genomics, proteomics, and in molecular biology in general. Similarly, methods for immobilization or attachment to surfaces and resins have become ubiquitous in sequencing, synthesis, analysis, and screening of oligonucleotides, peptides, and proteins. However, solid-phase tools have been employed to a much lesser extent in glycobiology and glycomics. This review provides a comprehensive overview of solid-phase chemical tools for glycobiology including methodologies and applications. We provide a broad perspective of different approaches, including some well-established ones, such as immobilization in microtiter plates and to cross-linked polymers. Emerging areas such as glycan microarrays and glycan sequencing, quantum dots, and gold nanoparticles for nanobioscience applications are also discussed. The applications reviewed here include enzymology, immunology, elucidation of biosynthesis, and systems biology, as well as first steps toward solid-supported sequencing. From these methods and applications emerge a general vision for the use of solid-phase chemical tools in glycobiology. PMID- 16716276 TI - N-Glycosyl-thiophene-2-carboxamides: synthesis, structure and effects on the growth of diverse cell types. AB - A range of N-glycosyl-thiophene-2-carboxamides, including a 6H-thieno[2,3 c]pyridin-7-one and a bivalent compound, have been synthesised and assayed for their effects on DNA synthesis in bovine aortic endothelial cells or on the growth of synoviocytes. Per-O-acetylated analogues of the glycoconjugates were significantly more effective inhibitors when compared to their corresponding non acetylated analogues, indicating that the lower potency observed for hydroxylated derivatives is due to less efficient transport of these compounds across the cell membrane. Thiophene-2-carboxamide was inactive as an inhibitor of bFGF induced proliferation, confirming the requirement of the carbohydrate residue for the observed biological properties. Glucose, mannose, galactose and 2-amino-2-deoxy glucose analogues were active as were a variety of substituted thiophene derivatives; the 6H-thieno[2,3-c]pyridin-7-one conjugate was inactive. Conformational analysis of the title compounds was investigated. X-ray crystal structural analysis of four N-glucosyl-thiophene-2-carboxamides showed that the pyranose rings adopted the expected 4C1 conformations and that Z-anti structures were predominant (H1-C1-N-H anomeric torsion angle varied from -168.2 degrees to 175.0 degrees ) and that the carbonyl oxygen and sulfur of the thiophene adopted an s-cis conformation in three of the isomers. In a crystal structure of a 3 alkynyl derivative, the hydrogen atom of the NH group was directed toward the acetylene group. The distance between the hydrogen atom and acetylene carbons and angles between nitrogen, hydrogen and carbon atoms were consistent with hydrogen bonding and this was supported by IR and NMR spectroscopic studies. The geometries of thiophene-2-carboxamides were explored by density functional theory (DFT) and Moller-Plesset (MP2) calculations and the s-cis conformer of thiophene 2-carboxamide was found to be more stable than its s-trans isomer by 0.83 kcal mol(-1). The s-cis conformer of 3-ethynyl-thiophene-2-carboxamide was 5.32 kcal mol(-1) more stable than the s-trans isomer. The larger stabilisation for the s cis conformer in the 3-alkynyl derivatives is explained to be due to a moderate hydrogen bonding interaction between the alkyne and NH group. PMID- 16716277 TI - Low- and high-resolution nuclear magnetic resonance (NMR) characterisation of hyaluronan-based native and sulfated hydrogels. AB - Hyaluronan-based hydrogels were synthesised using different crosslinking agents, such as 1,3-diaminopropane (1,3-DAP) and 1,6-diaminohexane (1,6-DAE). The hydrogels were sulfated to provide materials (Hyal-1,3-DAP, Hyal-1,6-DAE, HyalS 1,3-DAP and HyalS-1,6-DAE) that were characterised by both high- and low resolution nuclear magnetic resonance (NMR) spectroscopy. The (13)C NMR spectra of the materials were analysed to identify, characterise and study the crosslinking degree of the hydrogels. The crosslinking degree was also determined by potentiometric titration and the effectiveness of the two techniques was compared. Measurements of longitudinal relaxation times (spin-lattice) and of NOE enhancement were used to study the mobility of the hydrogels. Low-resolution NMR studies allowed the determination of the water transport properties in the hydrogels. In addition, the swelling degree for the various hydrogels was calculated as a function of the longitudinal and transversal relaxation times of the water molecules. Lastly, the self-diffusion coefficients of the water in interaction with the four polysaccharides were measured by the pulsed field gradient spin echo (PFGSE) sequence. PMID- 16716278 TI - Complete assignment of 1H and 13C NMR spectra of Gigartina skottsbergii lambda carrageenan using carrabiose oligosaccharides prepared by enzymatic hydrolysis. AB - lambda-Carrageenan extracted from Gigartina skottsbergii tetrasporophyte was completely digested by a purified Pseudoalteromonas carrageenovora lambda carrageenase. The main digestion products were fractionated and analysed by (1)H and (13)C NMR spectroscopy. All the oligosaccharides observed belong to the neo carrabiose oligosaccharide series indicating that the lambda-carrageenase cleaves the beta-(1-->4) glycosidic bonds. (1)H and (13)C NMR spectra recorded on oligomers from DP 2 to DP 8 were fully interpreted allowing unambiguous assignment of the lambda-carrageenan spectra. Besides the typical oligo-lambda carrageenans, we have also characterised a heptasulfated tetrasaccharide which demonstrates the random over-sulfation along the chain of G. skottsbergii lambda carrageenan. PMID- 16716279 TI - Penta-, hexa-, and heptasaccharide acceptor products of alternansucrase. AB - In the presence of suitable acceptor molecules, dextransucrase makes a homologous series of oligosaccharides in which the isomers differ by a single glucosyl unit, whereas alternansucrase synthesizes one trisaccharide, two tetrasaccharides, etc. For the example of maltose as the acceptor, if one considers only the linear, unbranched possibilities for alternansucrase, the hypothetical number of potential products increases exponentially as a function of the degree of polymerization (DP). Experimental evidence indicates that far fewer products are actually formed. We show that only certain isomers of DP >4 are formed from maltose in measurable amounts, and that these oligosaccharides belong to the oligoalternan series rather than the oligodextran series. When the oligosaccharide acceptor products from maltose were separated by size-exclusion chromatography and HPLC, only one pentasaccharide was isolated. Its structure was alpha-D-Glcp-(1-->6)-alpha-D-Glcp-(1-->3)-alpha-D-Glcp-(1-->6)-alpha-D-Glcp-(1- >4)-D-Glc. Two hexasaccharides were formed in approximately equal quantities: alpha-D-Glcp-(1-->3)-alpha-D-Glcp-(1-->6)-alpha-D-Glcp-(1-->3)-alpha-D-Glcp-(1- >6)-alpha-D-Glcp-(1-->4)-D-Glc and alpha-D-Glcp-(1-->6)-alpha-D-Glcp-(1-->6) alpha-D-Glcp-(1-->3)-alpha-D-Glcp-(1-->6)-alpha-D-Glcp-(1-->4)-D-Glc. Just one heptasaccharide was isolated from the reaction mixture, alpha-D-Glcp-(1-->6) alpha-D-Glcp-(1-->3)-alpha-D-Glcp-(1-->6)-alpha-D-Glcp-(1-->3)-alpha-D-Glcp-(1- >6)-alpha-D-Glcp-(1-->4)-D-Glc. We conclude that the enzyme is incapable of forming two consecutive alpha-(1-->3) linkages, and does not form products with more than two consecutive alpha-(1-->6) linkages. The distribution of products may be kinetically determined. PMID- 16716280 TI - The cardiac beta-adrenoceptors: Where and why? PMID- 16716281 TI - Membrane potassium currents in human radial artery and their regulation by nitric oxide donor. AB - OBJECTIVE: The human radial artery has demonstrated superior long-term results as a graft in coronary bypass surgery, but undesirable post-surgical spasm limits its clinical application. Few have examined its excitatory properties, especially the underlying ion channel mechanisms. In this study, we investigated the kinetic and pharmacological properties of the smooth muscle membrane potassium currents of this important artery. METHODS AND RESULTS: Using whole cell patch-clamp techniques, we found the K(+) current to be voltage-dependent and outwardly rectifying. Voltage-dependent inactivation was observed, being half-maximal at +28.0 mV but incomplete even at +40 mV. The K(+) currents were predominantly sensitive to the K(Ca) blocker tetraethylammonium (TEA; 63.9+/-12.1% inhibition, p<0.05), less sensitive to the Kv blocker 4-aminopyridine (4-AP; 32.8+/-4.4% inhibition, p<0.05), and the K(ATP) blocker glibenclamide (28.7+/-8.5% inhibition), at -20 mV testing potential. Resting membrane potential was -52.0+/ 6.8 mV (n=5), and suppression of K(+) currents by TEA and iberiotoxin (IbTx) caused membrane depolarization. Western blot analysis with channel-specific antibodies confirmed the presence of K(Ca) and Kv channel proteins. TEA evoked 20.7+/-9.9% of the contractile response to 60 mM KCl, whereas IbTx caused about 10% of the above response at 10(-7) M. The nitric oxide donor SNAP augmented membrane K(+) currents in a concentration-dependent fashion; the augmentation was completely suppressed by TEA, but was relatively insensitive to the guanylate cyclase inhibitor ODQ. CONCLUSIONS: The radial artery manifests mainly Ca(2+) dependent K(+) currents at rest; this current is augmented by nitric oxide through a cGMP- and protein kinase G-independent action. The relatively depolarized membrane potential, as well as its muscular structure, predisposes the radial artery to spasm. Agents that activate the Ca(2+)-dependent K(+) current could be of therapeutic value in preventing post-surgical vasospasm. PMID- 16716282 TI - Effects of isocoumarins isolated from Paepalanthus bromelioides on mitochondria: uncoupling, and induction/inhibition of mitochondrial permeability transition. AB - Isolated mitochondria may undergo uncoupling, and in presence of Ca(2+) at different conditions, a mitochondrial permeability transition (MPT) linked to protein thiol oxidation, and demonstrated by CsA-sensitive mitochondrial swelling; these processes may cause cell death either by necrosis or by apoptosis. Isocoumarins isolated from the Brazilian plant Paepalanthus bromelioides (Eriocaulaceae) paepalantine (9,10-dihydroxy-5,7-dimethoxy-1H naptho(2,3c)pyran-1-one), 8,8'-paepalantine dimer, and vioxanthin were assayed at 1-50 microM on isolated rat liver mitochondria, for respiration, MPT, protein thiol oxidation, and interaction with the mitochondrial membrane using 1,6 diphenyl-1,3,5-hexatriene (DPH). The isocoumarins did not significantly affect state 3 respiration of succinate-energized mitochondria; they did however, stimulate 4 respiration, indicating mitochondrial uncoupling. Induction of MPT and protein thiol oxidation were assessed in succinate-energized mitochondria exposed to 10 microM Ca(2+); inhibition of these processes was assessed in non energized organelles in the presence of 300 microM t-butyl hydroperoxide plus 500 microM Ca(2+). Only paepalantine was an effective MPT/protein thiol oxidation inducer, also releasing cytochrome c from mitochondria; the protein thiol oxidation, unlike mitochondrial swelling, was neither inhibited by CsA nor dependent on the presence of Ca(2+). Vioxanthin was an effective inhibitor of MPT/protein thiol oxidation. All isocoumarins inserted deeply into the mitochondrial membrane, but only paepalantine dimer and vioxantin decreased the membrane's fluidity. A direct reaction with mitochondrial membrane protein thiols, involving an oxidation of these groups, is proposed to account for MPT induction by paepalantine, while a restriction of oxidation of these same thiol groups imposed by the decrease of membrane fluidity, is proposed to account for MPT inhibition by vioxanthin. PMID- 16716283 TI - Resequencing the G6PT1 gene reveals a novel splicing mutation in a patient with glycogen storage disease type 1b. PMID- 16716284 TI - IFCC and Clinica Chimica Acta--50 years of partnership. PMID- 16716285 TI - Role of lipoic acid in reducing the oxidative stress induced by cyclosporine A. AB - BACKGROUND: Cyclosporine A (CsA) is the first choice immunosuppressant universally used for the prevention of allograft rejection in solid organ transplantation and immune-mediated diseases. However, with increasing use, evidence has accumulated that CsA therapy is associated with a variety of side effects, the most important being nephrotoxicity. We investigated the potential role of DL-alpha lipoic acid (LA), a universal antioxidant in combating the oxidative stress induced by CsA. METHODS: Adult male albino Wistar rats were divided into 4 treatment groups. Two groups received CsA (25 mg/kg body weight, orally for 21 days) to induce nephrotoxicity. One of these groups received LA treatment (20 mg/kg body weight, orally) for 21 days concurrently during CsA administration. A vehicle treated control group and a LA drug control were also included. RESULTS: CsA-induced nephrotoxicity was assessed in terms of increased activities of serum marker enzymes; alkaline phosphatase, acid phosphatase and lactate dehydrogenase. An apparent rise in the activities of N-acetyl-beta-D glucosaminidase, beta-glucuronidase and cathepsin D were seen in the renal tissue of CsA given rats, which were reversed upon treatment with LA. CsA administration induced significant elevation in lipid peroxidation along with abnormal levels of enzymic (superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione-S-transferase and glucose-6-phosphate dehydrogenase) and non-enzymic antioxidants (glutathione, vitamins C and E) in the rat kidney. LA administration improved renal function, by bringing about a significant decrease in peroxidative levels and increase in antioxidant status. CONCLUSION: These results indicate that LA has a protective action against CsA nephrotoxicity and suggest that the LA may find clinical application against a variety of toxins where cellular damage is a consequence of reactive oxygen species. PMID- 16716286 TI - Serum protein oxidation in patients with rheumatoid arthritis and effects of infliximab therapy. AB - OBJECTIVE: To examine protein oxidation in rheumatoid arthritis (RA) and evaluate its evolution after infliximab therapy in a subgroup of patients. METHODS: Seventy-one consecutive patients with RA were included. Among them, 30 patients refractory to conventional therapy were treated with infliximab. Serum markers of oxidative stress were determined at baseline and before the infusions of infliximab at weeks 6 and 30. Baseline values were compared with those in 30 healthy volunteers. RESULTS: Mean levels of serum carbonyl groups were significantly higher in RA patients than in controls (1.29+/-0.76 versus 0.58+/ 0.39 nmol/mg of protein, p<0.0001), whereas thiol levels were found to be lower (238.3+/-61.6 versus 316.5+/-54.8 micromol/L, p<0.0001). Thiol levels inversely correlated with the disease activity score (r=-0.42, p=0.004), and with CRP values (r=-0.45, p=0.001). Immunoblots showed that albumin and heavy chain immunoglobulin were oxidized more markedly than in healthy volunteers. Significantly lower levels of thiol groups were detected in patients with refractory RA disease (208.9+/-66.8 versus 264.2+/-43.0 micromol/L, p<0.0004) but concentrations of carbonyl groups were similar. Short-term treatment with infliximab significantly decreased carbonyl groups (0.97+/-0.47 nmol/mg protein, p=0.02) and increased thiol (231.2+/-48.7 micromol/L, p=0.02) levels. CONCLUSION: Our results highlight free radical protein damage in RA and a link with inflammation, as underlined by the beneficial effects of infliximab. PMID- 16716287 TI - Theoretical analysis on retention behavior of pigments in reversed-phase high performance liquid chromatographic (HPLC). AB - Quantitative structure-retention relationship (QSRR) models have been used successfully to predict and explain retention behavior of pigments in reversed phase high-performance liquid chromatography (HPLC). The semi-empirical quantum chemical method (PM3) in Gaussian98 was employed to calculate a set of molecular descriptors of pigments. Using multiple linear regression (MLR), we obtained empirical functions with high correlation coefficient between retention times and quantum-chemical descriptors. This analysis indicated that the proposed QSRR models were satisfactory. PMID- 16716288 TI - Wildfire air pollution and daily mortality in a large urban area. AB - Unusual air pollution episodes, such as when smoke from wildfires covers a large urban area, can be used to attempt to detect associations between short-term increases in particulate matter (PM) concentrations and subsequent mortality without relying on the sophisticated statistical models that are typically required in the absence of such episodes. The objective of this study was to explore whether acute increases in PM concentrations from wildfire smoke cause acute increases in daily mortality. The temporal patterns of daily nonaccidental deaths and daily cardiorespiratory deaths for June of 2002 in the Denver metropolitan area were examined and compared to those in two nearby counties in Colorado that were not affected by the wildfire smoke and to daily deaths in Denver in June of 2001. Abrupt increases in PM concentrations in Denver occurred on 2 days in June of 2002 as a result of wildfire smoke drifting over the Denver area. Small peaks in mortality corresponded to both of the PM peaks, but the first mortality peak also corresponded to a peak of mortality in the control counties, and cardiorespiratory deaths began to increase on the day before the second peak. Further, there was no detectable increase in cardiorespiratory deaths in the hours immediately following the PM peaks. Although the findings from this study do not rule out the possibility of small increases in mortality due to abrupt and dramatic increases in PM concentrations from wildfire smoke, in a population of over 2 million people no perceptible increases in daily mortality could be attributed to such events. PMID- 16716289 TI - Mink as a sentinel species in environmental health. AB - The concept of "sentinel species" is important in the environmental health sciences because sentinel species can provide integrated and relevant information on the types, amounts, availability, and effects of environmental contaminants. Here we discuss the use of mink (Mustela vison) as a sentinel organism by reviewing the pertinent literature from exposure- and effects-based studies. The review focuses on mercury (Hg) and polychlorinated biphenyls (PCBs), as they are persistent, ubiquitous, and bioaccumulative contaminants of concern to both humans and wildlife. Mink are widely distributed, abundant, and regularly trapped in temperate, aquatic ecosystems, and this makes them an excellent model to address issues in environmental pollution on both temporal and spatial scales. As a high-trophic-level, piscivorous mammal, mink can bioaccumulate appreciable concentrations of certain pollutants and have been shown to be sensitive to their toxic effects. The husbandry and life history of mink are well understood, and this has permitted controlled dosing experiments to be conducted using animals reared in captivity. These manipulative studies have yielded important quantitative information on exposure-response relationships and benchmarks of adverse health effects, and have also allowed the cellular mechanisms underlying toxic effects to be explored. Furthermore, the data accrued from the laboratory continue to validate observations made in the field. Research derived from mink can bridge and integrate multiple disciplines, and the information collected from this species has allowed environmental health scientists to better understand and characterize pollution effects on ecosystems. PMID- 16716290 TI - Blockade of cannabinoid CB1 receptors alters contextual learning and memory. AB - The endocannabinoid system appears to have an important role in specific aspects of learning and memory, yet there has been no systematic study of the role of cannabinoid receptors in contextual fear conditioning. The present study examined the effects of cannabinoid CB(1) receptor blockade on the acquisition, consolidation, and expression of contextual fear using the selective cannabinoid CB(1) receptor antagonist AM251. AM251 produced a decrease in the expression of contextual fear when administered prior to training, testing, or both. This effect was observed when footshock was signaled by an auditory cue but not in an unsignaled shock version of the task. Moreover, blocking cannabinoid CB(1) receptors had no effect on consolidation of contextual memory regardless of the conditioning paradigm. These data indicate that inhibition of cannabinoid CB(1) receptors produces specific deficits in processing contextual information and that the effects of CB(1) antagonists on contextual learning may differ from effects on other types of learning. PMID- 16716291 TI - ATP stimulates interleukin-6 production via P2Y receptors in human HaCaT keratinocytes. AB - We evaluated the role of ATP in functions of human HaCaT keratinocytes. ATP was released from HaCaT cells by changing the culture medium. Reverse transcription polymerase chain reaction analysis revealed that HaCaT cells expressed multiple P2 purinergic receptor mRNAs. UTP was the most potent agonist to increase the intracellular Ca2+ concentration ([Ca2+]i). UTP and ATP caused the accumulation of [3H]inositol phosphates, suggesting that UTP binds to the Gq/11-coupled P2Y receptor. UTP increased IL-6 mRNA and protein levels, and the increases were inhibited by a P2 purinergic receptor antagonist (suramin, 300 microM). While a protein kinase C inhibitor (GF109203X, 10 microM) was without effect, an intracellular free Ca2+ chelator (BAPTA-AM, 50 microM) suppressed UTP-mediated IL 6 induction. These results suggest that 1) ATP is released from HaCaT cells upon physical stimulation and may act as an autocrine molecule, and 2) the stimulation of P2Y receptors causes IL-6 production via mRNA expression through [Ca2+]i elevation. PMID- 16716292 TI - Angiotensin II potentiates inflammatory edema in rats: Role of mast cell degranulation. AB - The aim of this study was to evaluate the effect of angiotensin II on models of acute inflammation. This study shows that angiotensin II potentiates the carrageenan- and dextran-induced paw edema. The administration of angiotensin II does not change the myeloperoxidase activity, neither the tissue content of interleukin-1 beta and tumor necrosis alpha nor the neutrophil migration to the peritoneal cavity, but induces significant enhancement of mast cell degranulation. The anti-histamine, mepyramine, and the anti-serotonin, metisergyde, reduce the angiotensin II-facilitated dextran-induced edema. Our results suggest that angiotensin II increases the vascular permeability through induction of mast cell degranulation and that this effect is mediated by the angiotensin AT2 receptor, since the angiotensin AT1 receptor antagonist and the angiotensin AT2 receptor agonist potentiated the paw edema. PMID- 16716293 TI - Ketamine enhances the expression of serine racemase and D-amino acid oxidase mRNAs in rat brain. AB - We have evaluated the effects of the acute administration of noncompetitive N methyl-D-aspartate receptor antagonist, ketamine, on the expression of serine racemase and D-amino acid oxidase mRNAs in several brain areas of rats. The ketamine administration produced a dose-dependent and transient elevation in the levels of serine racemase and D-amino acid oxidase mRNAs in all the brain areas. These findings suggest that there is a relationship between the gene expression of the d-serine-related enzymes and the blockade of the N-methyl-D-aspartate receptors. PMID- 16716294 TI - Alpha1L-adrenoceptors mediate contractions of the isolated mouse prostate. AB - The subtype of alpha1-adrenoceptor mediating noradrenaline-induced contractile responses in isolated mouse prostate glands was investigated. Adrenoceptor agonists were able to produce concentration-dependent contractions with the following rank order of potency: adrenaline > or = noradrenaline > or = clonidine = phenylephrine > dopamine > or = isoprenaline. Concentration-response curves to noradrenaline of the prostatic smooth muscle were antagonised by prazosin, N-[2 (2-cyclopropylmethoxyphenoxy)ethyl]-5-chloro-alpha, alpha-dimethyl-1H-indole-3 ethanamine (RS-17053), 2-(2,6-dimethoxyphenoxyethyl)aminomethyl-1,4-benzodioxane (WB 4101), tamsulosin and yohimbine with mean antagonist affinity estimates (pA2 or apparent pKB) of 8.12+/-0.10, 6.56+/-0.11, 8.38+/-0.06, 10.14+/-0.19 and 7.38+/-1.36 respectively. Propranolol (1 microM) had no antagonist activity (P = 0.994, n = 6). Yohimbine (0.01, 0.1, 1 microM) had no antagonist activity in the presence of prazosin (0.1 microM) (P > or = 0.059). The results obtained indicate that alpha1-adrenoceptors mediate the contractile response in isolated preparations of the mouse prostate. Furthermore, the particular subtype of alpha1 adrenoceptor mediating the response to exogenously administered noradrenaline corresponds to the alpha1L-subtype, the same subtype as that which has been shown to mediate noradrenaline-induced contractile activity in the human prostate. PMID- 16716295 TI - Upstream signaling of protein kinase C-epsilon in xenon-induced pharmacological preconditioning. Implication of mitochondrial adenosine triphosphate dependent potassium channels and phosphatidylinositol-dependent kinase-1. AB - Xenon elicits preconditioning of the myocardium via protein kinase C-epsilon. We determined the implication of (1) the mitochondrial adenosinetriphosphate dependent potassium (K(ATP)) channels and (2) the 3'phosphatidylinositol dependent kinase-1 (PDK-1) in activating protein kinase C-epsilon. For infarct size measurements, anaesthetized rats were subjected to 25 min of coronary artery occlusion followed by 120 min of reperfusion. Rats received xenon 70% during three 5-min periods before ischaemia with or without the K(ATP) channel blocker 5 hydroxydecanoate or Wortmannin as PI3K/PDK-1 inhibitor. For Western blot, hearts were excised at five time points after xenon preconditioning (Control, 15, 25, 35, 45 min). Infarct size was reduced from 42+/-6% (mean+/-S.D.) to 27+/-8% after xenon preconditioning (P<0.05). Western blot revealed an increased activation of PKC-epsilon after 45 min and of PDK-1 after 25 min during xenon preconditioning. 5-hydroxydecanoate and Wortmannin blocked both effects. PKC-epsilon is activated downstream of mitochondrial K(ATP) channels and PDK-1. Both pathways are functionally involved in xenon preconditioning. PMID- 16716296 TI - Identification of stem cells from human umbilical cord blood with embryonic and hematopoietic characteristics. AB - We identified stem cells from the umbilical cord blood, designated cord blood stem cells (CB-SC). CB-SC displayed important embryonic stem (ES) cell characteristics including expression of ES-cell-specific molecular markers including transcription factors OCT-4 and Nanog, along with stage-specific embryonic antigen (SSEA)-3 and SSEA-4. CB-SC also expressed hematopoietic cell antigens including CD9, CD45 and CD117, but were negative for CD34. CB-SC displayed very low immunogenicity as indicated by expression of a very low level of major histocompatibility complex (MHC) antigens and failure to stimulate the proliferation of allogeneic lymphocytes. CB-SC could give rise to cells with endothelial-like and neuronal-like characteristics in vitro, as demonstrated by expression of lineage-associated markers. Notably, CB-SC could be stimulated to differentiate into functional insulin-producing cells in vivo and eliminated hyperglycemia after transplantation into a streptozotocin-induced diabetic mouse model. These findings may have significant potential to advance stem-cell-based therapeutics. PMID- 16716297 TI - Reversible, activity-dependent targeting of profilin to neuronal nuclei. AB - The actin cytoskeleton in pyramidal neurons plays a major role in activity dependent processes underlying neuronal plasticity. The small actin-binding protein profilin shows NMDA receptor-dependent accumulation in dendritic spines, which is correlated with suppression of actin dynamics and long-term stabilization of synaptic morphology. Here we show that following NMDA receptor activation profilin also accumulates in the nucleus of hippocampal neurons via a process involving rearrangement of the actin cytoskeleton. This simultaneous targeting to dendritic spines and the cell nucleus suggests a novel mechanism of neuronal plasticity in which profilin both tags activated synapses and influences nuclear events. PMID- 16716298 TI - Human astrocytes can be induced to differentiate into cells with neuronal phenotype. AB - Several recent studies have proposed that astrocytes may contribute to neurogenesis, not only as a source of trophic substances regulating it, but also as stem cells themselves. In order to better understand these mechanisms, primary astrocyte cultures were established from human fetal brain. After 3-4 weeks in culture, astrocytes (about 95% GFAP+; neurofilament, NF-; neuro-specific enolase, NSE-) were treated with a cocktail of protein kinase activators and FGF-1. After 5 h of treatment, most cells showed morphological changes that increased progressively up to 24-48 h, exhibiting a round cell body with long processes. Immunocytochemistry showed that treatment-induced NF and NSE expression in about 40% of cells. Nestin expression increased after treatment, whereas GFAP immunostaining was not significantly modified. Western blot and RT-PCR confirmed the results. No neuronal electrophysiological properties were observed after treatment, suggesting an incomplete maturation under these experimental conditions. Understanding the regenerative capability and neurogenic potential of astrocytes might be useful in devising therapeutic approaches for a variety of neurological disorders. PMID- 16716299 TI - Nerve growth factor effect on human primary fibroblastic-keratocytes: possible mechanism during corneal healing. AB - In response to corneal injury, cytokines and growth factors play a crucial role by influencing epithelial-stromal interaction during the healing and reparative processes which may resolve in tissue remodeling and fibrosis. While transforming growth factor-beta1 (TGF-beta1) is considered the main profibrogenic modulator of these process, recently the nerve growth factor (NGF) appears as a pleiotropic modulator of wound-healing and inflammatory responses. Interestingly in the cornea, where NGF, trkA(NGFR) and p75(NTR) are expressed by epithelial cells and keratocytes, the NGF eye-drop induces the healing of neurotrophic or autoimmune corneal ulcers. During corneal healing, quiescent keratocytes are replaced by active fibroblast-like keratocytes/myofibroblasts. While the NGF effect on epithelial cells has been investigated, no data are reported for NGF effects on fibroblastic-keratocytes, during corneal healing. NGF, trkA(NGFR) and p75(NTR) were found expressed by fibroblastic-keratocytes. NGF was able to induce fibroblastic-keratocyte differentiation into myofibroblasts, migration, Metalloproteinase-9 expression/activity and contraction of a 3D collagen gel, without affecting their proliferation and collagen production. These data also show a two-directional control of fibroblastic-keratocytes by NGF and TGF-beta1. To sum up, the findings of this study indicate that NGF can modulate some functional activities of fibroblastic-keratocytes, thus substantiating the healing effects of NGF on corneal wound-healing. PMID- 16716300 TI - The study of c-Src kinase and pStat3 protein expression in retinoblastoma. AB - We examine the immunoreactivity of the non-receptor tyrosine kinase, c-Src kinase and its downstream molecule, signal transducer and activator of transcription 3 (pStat3) in retinoblastoma (RB), and correlation with invasiveness and differentiation. Tumor samples from 40 patients with RB were available for the study. There were 18 tumors in group 1 (non-invasive) and 22 tumors in group 2 (invasive). The immunoreactivity of c-Src kinase and pStat3 was compared in the two groups of tumors. Group 1 (non-invasive) RB showed intermediate c-Src kinase immunoreactivity (Allred score 4-5) in 14/18 tumors and low immunoreactivity (Allred score 2-3) in 4/18 tumors. pStat3 was intermediate (Allred score 4-5) in 6/18 tumors and negative (Allred score 0) in 12/18 tumors. Group 2 (invasive) RB showed high c-Src kinase immunoreactivity (Allred score 6-8) in 22/22 tumors and high pStat3 (Allred score 6-8) in 19/22 tumors. The expression of c-Src kinase (P<0.001) and pStat3 (P<0.001) was significantly higher in group 2 RB. Src kinase expression (P<0.05) and pStat3 expression (P<0.05) was higher in the poorly differentiated tumors compared to moderately- and well-differentiated tumors. The increased expression of c-Src kinase and pStat3 expression could play a role in the invasiveness of group 2 tumors. Further characterization of the pathways involved in the pathogenesis of RB will shed light on fundamental mechanisms of tumorigenesis. PMID- 16716301 TI - Leishmania donovani: an in vitro study of antimony-resistant amphotericin B sensitive isolates. AB - Drug sensitivity of clinically antimony-unresponsive Leishmania donovani isolates from Eastern Sudan was evaluated in an in vitro culture system against sodium stibogluconate (Pentostam) and Amphotericin B. Eight isolates, six from antimony resistant and two from clinically responsive patients were included in the study. Parasites were tested as promastigotes and four of them were selected to be tested as amastigotes using a murine macrophage-like cell line. The results indicated that the conventional promastigotes and amastigotes-screening assays did not correlate with the clinical picture of patients. In vivo unresponsiveness does not necessarily mean primary parasite resistance. Amphotericin B could be a suitable second line drug in patients unresponsive to pentostam and without concomitant diseases, if close hospital monitoring is available. Promastigotes sensitivity testing concentrations are virtually incomparable with the in vivo clinically curable doses and the amastigotes/macrophage test concentrations. PMID- 16716302 TI - Analysis of mosquito bloodmeals using RFLP markers. AB - An important variable in the amplification of arthropod vector-borne diseases is the degree of contact between human hosts and mosquito vectors. To analyze this interaction, a DNA based method was developed to differentiate human bloodmeals from other sources in the mosquito Anopheles stephensi (Diptera: Culicidae) Liston. A portion of the host mitochondrial DNA cytochrome B genes were PCR amplified and classified to the species level based on their restriction fragment length polymorphism (RFLP). The cytochrome B sequences showed sufficient interspecific polymorphism to distinguish between human, cow, sheep, chicken, and guinea pig hosts. XhoI could distinguish human from other vertebrates whereas TaqI alone could separate the others. The importance of these results in epidemiological studies of malaria and other vector borne diseases is discussed. PMID- 16716303 TI - Pathology of Tnf-deficient mice infected with Plasmodium chabaudi adami 408XZ. AB - Tumor necrosis factor alpha (Tnf) plays a pleiotropic role in murine malaria. Some investigations have correlated Tnf with hypothermia, hyperlactatemia, hypoglycemia, and a suppression of the erythropoietic response, although others have not. In this study, we have evaluated parasitemia, survival rate and several pathological features in C57BL/6JTnf(-/-) and C57BL/6JTnf(+/+) mice after infection with Plasmodium chabaudi adami 408XZ. Compared to the C57BL/6JTnf(+/+) mice, C57BL/6JTnf(-/-) mice showed increased parasitemia and decreased survival rate, whereas blood glucose, blood lactate and body weight were not significantly different. However, C57BL/6JTnf(-/-) mice suffered significantly more from severe anemia and hypothermia than C57BL/6JTnf(+/+) mice. These results suggest that Tnf is an important mediator of parasite control, but not of anemia development. We hypothesize that the high mortality observed in the Tnf knock-out mice is due to increased anemia and pathology as a direct result of increased levels of parasitemia. PMID- 16716304 TI - An essential role of Cdc42-like GTPases in mitosis of HeLa cells. AB - Here we used RNA interference and examined possible redundancy amongst Rho GTPases in their mitotic role. Chromosome misalignment is induced significantly in HeLa cells by Cdc42 depletion and not by depletion of either one or all of the other four Cdc42-like GTPases (TC10, TCL, Wrch1 or Wrch2), four Rac-like GTPases or three Rho-like GTPases. Notably, combined depletion of Cdc42 and all of the other four Cdc42-like GTPases significantly enhances chromosomal misalignment. These observations suggest that Cdc42 is the primary GTPase functioning during mitosis but that the other four Cdc42-like GTPases can also assume the mitotic role in its absence. PMID- 16716305 TI - Elevated phenylalanine levels interfere with neurite outgrowth stimulated by the neuronal cell adhesion molecule L1 in vitro. AB - Elevated levels of phenylalanine (Phe) as observed in patients with phenylketonuria interfere with proper neuronal development, leading to severe psychomotor deficits and mental retardation. We have analyzed the effects of Phe on neurite outgrowth in vitro. When expressed in fibroblasts, the neuronal cell adhesion molecules L1 and plexin B3 strongly increase the length of neurites emanating from cerebellar neurons in co-culture experiments. Elevated Phe blocks L1-mediated, but not plexin B3-mediated outgrowth, whereas tyrosine is ineffective. Elevated Phe also interferes with aggregation of fibroblasts overexpressing L1, suggesting that the pathological effect of elevated Phe occurs by interfering with L1-mediated cell adhesion. PMID- 16716306 TI - Regulation of gonadotropin secretion and puberty onset by neuromedin U. AB - Neuromedin U (NMU), an anorexigenic peptide, was originally isolated from porcine spinal cord in 1985. As NMU is abundant in the anterior pituitary gland, we investigated the effects of NMU on gonadotropin secretion. Both NMU and its receptors, NMUR1 and NMUR2, were expressed in the pituitary gland. NMU suppressed LH and FSH releases from rat anterior pituitary cells. Moreover, NMU-deficient mice exhibit an early onset of vaginal opening. The LHbeta/FSHbeta ratio, which is an index of puberty onset, is high in young NMU-deficient mice. These results indicate that NMU suppresses gonadotropin secretion and regulates the onset of puberty. PMID- 16716307 TI - In vitro inhibition of transthyretin aggregate-induced cytotoxicity by full and peptide derived forms of the soluble receptor for advanced glycation end products (RAGE). AB - Familial amyloidotic polyneuropathy is a neurodegenerative disorder characterized by systemic extracellular deposition of transthyretin (TTR) amyloid fibrils. The latter have been proposed to trigger neurodegeneration through engagement of the receptor for advanced glycation end products (RAGE). Here we show that TTR interaction with RAGE is conserved across mouse and human species and is not dependent on RAGE glycosylation. Moreover, strand D of TTR structure seems important for the TTR-RAGE interaction as well as a motif in RAGE (residues 102 118) located within the V-domain; this motif suppressed TTR aggregate-induced cytotoxicity in cell culture. PMID- 16716308 TI - CHOP is involved in neuronal apoptosis induced by neurotrophic factor deprivation. AB - Neurotrophic factors are essential for the survival of neurons. We found that the endoplasmic reticulum (ER) stress-C/EBP homologues protein (CHOP) pathway to be activated during neurotrophic factor deprivation-induced apoptosis in PC12 neuronal cells and in primary cultured neurons, and this apoptosis was suppressed in the neurons from chop(-/-) mice. In addition, we found that CHOP is expressed in the subventricular zone (SVZ) and striatum of the young adult mouse brain. The number of apoptotic cells in the SVZ decreased in chop(-/-) mice. These results indicate that the ER stress-CHOP pathway plays a role in neuronal apoptosis during the development of the brain. PMID- 16716309 TI - Hyperosmotic stress activates p65/RelB NFkappaB in cultured cardiomyocytes with dichotomic actions on caspase activation and cell death. AB - NFkappaB is a participant in the process whereby cells adapt to stress. We have evaluated the activation of NFkappaB pathway by hyperosmotic stress in cultured cardiomyocytes and its role in the activation of caspase and cell death. Exposure of cultured rat cardiomyocytes to hyperosmotic conditions induced phosphorylation of IKKalpha/beta as well as degradation of IkappaBalpha. All five members of the NFkappaB family were identified in cardiomyocytes. Analysis of the subcellular distribution of NFkappaB isoforms in response to hyperosmotic stress showed parallel migration of p65 and RelB from the cytosol to the nucleus. Measurement of the binding of NFkappaB to the consensus DNA kappaB-site binding by EMSA revealed an oscillatory profile with maximum binding 1, 2 and 6h after initiation of the hyperosmotic stress. Supershift analysis revealed that p65 and RelB (but not p50, p52 or cRel) were involved in the binding of NFkappaB to DNA. Hyperosmotic stress also resulted in activation of the NFkappaB-lux reporter gene, transient activation of caspases 9 and 3 and phosphatidylserine externalization. The effect on cell viability was not prevented by ZVAD (a general caspase inhibitor). Blockade of NFkappaB with AdIkappaBalpha, an IkappaBalpha dominant negative overexpressing adenovirus, prevented activation of caspase 9 (more than that caspase 3) but did not affect cell death in hyperosmotically stressed cardiomyocytes. We conclude that hyperosmotic stress activates p65 and RelB NFkappaB isoforms and NFkappaB mediates caspase 9 activation in cardiomyocytes. However cell death triggered by hyperosmotic stress was caspase- and NFkappaB-independent. PMID- 16716310 TI - Purification of the skeletal muscle protein Endopin 1B and characterization of the genes encoding Endopin 1A and 1B isoforms. AB - In the present work, a new endopin-like serpin designed mEndopin 1B was purified from bovine muscle. Biochemical characterizations (amino acid sequencing and Maldi-Tof mass spectrometry peptide mapping) demonstrated that the purified protein is different from the previously described Endopin 1, renamed mEndopin 1A. The genes and cDNA of both endopins were characterized. The cDNA sequence of mEndopin 1B encodes a predicted protein of 411 amino-acids with a molecular mass of 43808Da. The mEndopin 1B gene comprised four coding exons and an additional 5' untranslated exon. The reactive site sequence of mEndopin 1B is somewhat different from that of mEndopin 1A. Nevertheless, both serpins have a similar peptidase inhibitory pattern against examined proteases (elastase, trypsin, plasmin and chymotrypsin). The high expression of both mEndopin 1A and 1B in bovine serum and tissues and their high efficiency to inhibit elastase (k(ass) approximately 10(6)-10(7) M(-1) s(-1)) suggested that these serpins might play a major role in inflammatory processes. PMID- 16716311 TI - Frozen-thawed embryo transfer and live birth: Long-term follow-up after one oocyte retrieval. AB - Frozen-thawed embryos accounted for 39% (249 of 639) of live births from 931 consecutive first oocyte retrievals after median follow-up of 6.5 years with consistent use of pronuclear-stage freezing and cleavage-stage transfer. Survival after thaw was 95% (2,129 of 2,247). Implantation and live birth rates per individual frozen-thawed embryo transfered were 22% (431 of 1,937) and 18% (346 of 1,937), respectively. PMID- 16716312 TI - Basal and cycle androgen levels correlate with in vitro fertilization stimulation parameters but do not predict pregnancy outcome. AB - OBJECTIVE: To evaluate androgen levels before and during IVF. To assess for an association between androgen levels and IVF stimulation parameters or IVF pregnancy outcome. DESIGN: Prospective cohort study. SETTING: Residency-based IVF program. PATIENT(S): One hundred seventeen infertility patients. INTERVENTION(S): Androgen levels were evaluated on basal day 3 and during the IVF stimulation cycle. MAIN OUTCOME MEASURE(S): Pregnancy outcome rates and IVF stimulation parameters. RESULT(S): Mean serum androgen levels did not differ among different pregnancy outcomes. Multiple linear regression analysis revealed that body mass index (BMI) and oocyte number had a significant positive association with basal testosterone levels. Mean ovarian volume correlated negatively and follicle number correlated positively with testosterone levels on day 6 of stimulation. Peak E(2) and BMI correlated positively with testosterone on day of hCG administration. The interval change in androgen levels throughout the IVF cycle was not associated with outcome rates. Likewise, threshold analysis did not reveal any significant androgen level that affected pregnancy outcome. CONCLUSION(S): Serum androgen levels during IVF correlate with IVF stimulation parameters. However, these data do not support an influence of serum androgen levels on IVF pregnancy outcome rates. PMID- 16716313 TI - Deciding the fate of supernumerary frozen embryos: a survey of couples' decisions and the factors influencing their choice. AB - OBJECTIVE: To investigate the decisions that couples make regarding supernumerary frozen embryos, the factors influencing these decisions, and the degree of difficulty involved in reaching a decision; and to canvass attitudes toward donating embryos to stem-cell research. DESIGN: Anonymous postal survey. SETTING: A large, private IVF clinic in a major city in Victoria, Australia. PATIENT(S): A consecutive cohort of couples who contacted the Monash IVF clinic in relation to embryos in long-term storage. INTERVENTION(S): Subjects completed a survey regarding decisions about surplus frozen embryos. MAIN OUTCOME MEASURE(S): Couples' decision regarding supernumerary embryos and reasons for the decision, experience of deciding, and attitudes about embryo donation for stem-cell research. RESULT(S): Forty percent (123/311) returned completed questionnaires. The most common decision was donation to research (42%). Altruistic motives and desire not to waste embryos were determinants of embryo donation. Determinants of disposal were not wanting a full sibling to existing children and opposition of embryo research. Forty-five percent found deciding distressing. The majority (69%) approved of embryo donation to stem-cell research. CONCLUSION(S): Most couples preferred embryos to come to some use rather than being disposed of. Almost half the sample reported finding the decision making distressing. A majority approved of embryo donation for stem-cell research. PMID- 16716314 TI - Embryo quality is more important for younger women whereas age is more important for older women with regard to in vitro fertilization outcome and multiple pregnancy. AB - OBJECTIVE: To determine the efficiency of embryo scoring systems for multiple pregnancy in women undergoing IVF procedures. DESIGN: Retrospective record analysis. SETTING: University hospital, tertiary medical center. PATIENT(S): Three hundred one patients undergoing controlled ovarian stimulation, IVF/intracytoplasmic sperm injection (ICSI), and day 3 embryo transfer. INTERVENTION(S): IVF/ICSI and embryo transfer. MAIN OUTCOME MEASURE(S): Rate of pregnancy and rate of multiple pregnancy. RESULT(S): The score of the best three embryos (Top3) was more correlated with IVF outcome than were the number of good embryos (P=.009) or the cumulative embryo score (P=.038). In the logistic regression model, Top3 was more relevant to IVF outcome and multiple pregnancy for younger patients than was age (P<.05). For older patients, age was more correlated with IVF outcome and multiple pregnancy than was embryo morphology (P<.05). CONCLUSION(S): The embryo morphology criteria can help reduce the number of embryos transferred into younger patients. We could use the age of patients as an indicator to determine the number of embryos transferred into older patients. PMID- 16716315 TI - Pregnancy outcome in patients with early recurrent abortion following laparoscopic tubal corneal interruption of a fallopian tube with hydrosalpinx. AB - OBJECTIVE: To investigate the outcome of pregnancy following laparoscopic unilateral tubal fulguration of hydrosalpinges in patients with recurrent spontaneous early abortion. DESIGN: Prospective randomized controlled trial. SETTING: University tertiary center. PATIENT(S): Thirteen patients with history of unexplained recurrent early spontaneous abortion and a unilateral hydrosalpinx diagnosed by sonography and hysterosalpingography in whom other causes of abortion were excluded. INTERVENTION(S): The treatment group (group I) consisted of seven women who underwent laparoscopic unilateral tubal fulguration. The control group (group II) consisted of six patients for whom no surgical intervention was performed. MAIN OUTCOME MEASURE(S): Continuation of pregnancy over first trimester. RESULT(S): Six patients in the treatment group and five in the control group conceived. Five patients in the treatment group and none in the control group had pregnancy beyond the first trimester and finally reached 36-40 weeks gestational age (P=.02). CONCLUSION(S): Laparoscopic tubal fulguration improves pregnancy outcome in selected patients with previous recurrent early abortion and a unilateral hydrosalpinx. PMID- 16716316 TI - The presence of ovarian endometriomas is associated with a reduced responsiveness to gonadotropins. AB - OBJECTIVE: To clarify whether the presence of ovarian endometriomas is associated with a reduced responsiveness to ovarian hyperstimulation. DESIGN: Observational study. SETTING: University teaching hospital. PATIENT(S): Women selected for in vitro fertilization-intracytoplasmic sperm injection (IVF-ICSI) cycles who were found to have unilateral ovarian endometriomas and who did not undergo previous ovarian surgery. INTERVENTION(S): Ovarian hyperstimulation using gonadotropins. MAIN OUTCOME MEASURE(S): Number of codominant follicles (mean diameter >15 mm) in the affected and in the contralateral intact gonads. RESULT(S): Thirty-six patients were enrolled. They underwent 56 IVF-ICSI cycles. The number of codominant follicles in the intact and affected ovaries were 4.0 +/- 2.2 and 3.0 +/- 1.7, respectively (P=.01). This difference corresponded to a mean reduction (95% confidence interval [CI]) of 25% (6%-44%). This deleterious effect was more evident in women with larger cysts, in those with more than one cyst, and in those who were more responsive to ovarian hyperstimulation. CONCLUSION(S): The presence of ovarian endometriomas is associated with a reduced responsiveness to gonadotropins. PMID- 16716317 TI - ProteinChip technology is a useful method in the pathogenesis and diagnosis of endometriosis: a preliminary study. AB - OBJECTIVE: To test the feasibility of ProteinChip (Ciphergen Biosystems, Inc., Fremont, CA) technology as a proteomic tool in discovering and identifying proteins that are differentially expressed in endometrium, endometriotic tissue, and normal peritoneum from women with and without endometriosis. DESIGN: Differential analysis of protein expression in women with and without endometriosis. SETTING: University hospital. PATIENT(S): A total of nine patients during their secretory phase (days 20-22) were selected for this study on the basis of cycle phase and presence/or absence of endometriosis. INTERVENTION(S): Twelve tissues used in the study included six endometrial biopsies from women with mild endometriosis (n = 3) and a normal pelvis (n = 3) as well as paired samples of peritoneal endometriotic lesions (n = 3) and macroscopically normal peritoneum biopsies (n = 3) from three women with endometriosis. MAIN OUTCOME MEASURE(S): Numerous expression differences were observed in the above comparisons, representing both up-regulation and down-regulation in protein and peptide expression levels. RESULT(S): Endometrial expression for a number of proteins and peptides in the range of 2.8-12.3 kDa was 3-24 times lower in women with endometriosis than in those without endometriosis. When compared with normal peritoneum, endometriotic lesions showed an increased expression for a set of proteins and peptides in the range of 3-96 kDa, and especially an up-regulated cluster of proteins between 22 and 23 kDa, identified to be transgelin, a smooth muscle actin-binding protein. CONCLUSION(S): This preliminary study demonstrated that differential protein profiling by using ProteinChip array technology is feasible, reproducible, and may be developed into a powerful tool for endometriosis research. PMID- 16716318 TI - Instability in the transmission of the myotonic dystrophy CTG repeat in human oocytes and preimplantation embryos. AB - OBJECTIVE: To elucidate the timing and variability of CTG repeat expansion within the human dystrophia myotonica protein kinase (DMPK) gene in early development. DESIGN: Triplet-primed polymerase chain reaction was used to amplify the expanded CTG repeat in oocytes and embryos obtained from myotonic dystrophy 1 (DM1) patients, and a heminested polymerase chain reaction approach was used to amplify the normal CTG repeats in supernumerary IVF embryos. SETTING: University hospital laboratory. PATIENT(S): Two DM1-affected females undergoing preimplantation genetic diagnosis who carried different CTG repeats. Also, 61 IVF patients who carried a (CTG)(5-18) and (CTG)(19-37) normal DMPK repeat. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): The degree of expansion of the repeat in the oocytes and embryos compared with the DM1-affected maternal repeat and the size of the (CTG)(19-37) repeat compared with the parental size in IVF embryos. RESULT(S): The degree of repeat expansion was greater than the DM1 maternal lymphocyte for two of four oocytes, including a germinal vesicle-stage oocyte and 17 of 20 three cell to blastocyst stage embryos. A change in the (CTG)(19-37) repeat was seen in 7 (7%) of 95 paternal transmissions but in no maternal transmissions. CONCLUSION(S): Because the repeat was already expanded in the immature oocyte, the initial expansion most likely occurs during oogenesis. A variable degree of DMPK(CTG)(n) expansion in the embryo is seen from different mothers. In addition, instability in paternal transmission of normal-range (CTG)(19-37) repeats occurs at the level of the embryo. PMID- 16716319 TI - Use of laser-assisted intracytoplasmic sperm injection (ICSI) in patients with a history of poor ICSI outcome and limited metaphase II oocytes. AB - In this study, laser-assisted intracytoplasmic sperm injection (ICSI) improved the fertilization rate and the embryo quality in patients with a history of poor ICSI outcome and with limited metaphase II oocytes. This technique is less traumatic to the oocytes during the procedure, and the use of the technique may be expanded. PMID- 16716320 TI - Transvaginal oocyte retrieval for in vitro fertilization complicated by ovarian abscess during pregnancy. AB - OBJECTIVE: To describe an ovarian abscess presenting very late after oocyte retrieval for IVF with several unusual clinical features. DESIGN: Case report. SETTING: Academic medical center. PATIENT(S): A 35-year-old nulliparous woman underwent IVF with uncomplicated transvaginal oocyte retrieval (TVOR), resulting in a dizygotic twin pregnancy. At 13 weeks of pregnancy she presented with vaginal discharge, but was otherwise constitutionally well. At 30 weeks she developed a low-grade fever, and the diagnosis of ovarian abscess was made. She received broad-spectrum antibiotics, and the abscess was drained percutaneously after cesarean delivery of twins. INTERVENTION(S): Antimicrobial therapy; cesarean section; postpartum percutaneous drainage. MAIN OUTCOME MEASURE(S): Clinical and radiologic resolution of infection. RESULT(S): Complete resolution of the abscess; delivery of healthy twins. CONCLUSION(S): Infectious complications of TVOR and other surgical procedures usually occur within days of the intervention. Our case illustrates the possibility of infectious complications of TVOR presenting months after the procedure. Our patient did not become acutely ill due to the formation of a spontaneous vaginal fistula, which allowed the abscess to drain. The optimal management of this complication is unclear, but final resolution of any pelvic abscess generally requires drainage. PMID- 16716321 TI - A combination treatment of prednisone, aspirin, folate, and progesterone in women with idiopathic recurrent miscarriage: a matched-pair study. AB - OBJECTIVE: To compare a combination treatment of prednisone, aspirin, folate, and progesterone with no treatment in women with idiopathic recurrent miscarriage (IRM). DESIGN: Matched-pair study. SETTING: Academic research institution. SUBJECT(S): Women with a history of IRM, defined as three or more consecutive miscarriages before 20 weeks' gestation without associated anatomic, cytogenetic, hormonal, and infectious pathologies or antiphospholipid syndrome. INTERVENTION(S): Eighty of 210 eligible women consented to participate and were treated with prednisone (20 mg/d) and progesterone (20 mg/d) for the first 12 weeks of gestation, aspirin (100 mg/d) for 38 weeks of gestation, and folate (5 mg every second day) throughout their pregnancies. Fifty of 80 women became pregnant; they were compared with 52 women with IRM (matched for age and number of miscarriages), who became pregnant without treatment during the same observation period. MAIN OUTCOME MEASURE(S): Live birth rate, complications of pregnancy, such as preeclampsia, premature birth, and intrauterine growth restriction, and therapy-related side effects. RESULT(S): The overall live birth rates of the treatment and control groups were 77% (40 of 52) and 35% (18 of 52) (P=.04). The rates of first and second trimester miscarriage among the treatment and control groups were 19% (10 of 52) and 0 (0 of 52), and 63% (33 of 52) and 2% (1 of 52), respectively (P=.09 and P=1.0, respectively). The median gestational age at birth and median birth weight did not differ between the groups. We observed two and three cases of premature birth among the treatment and control groups, respectively (P=.3) and no cases of intrauterine growth restriction and Cushing's disease. Of 80 women who started treatment, one woman had an ectopic pregnancy and one woman terminated her pregnancy due to fetal chromosome aberration (trisomy 18). Three women stopped treatment due to nausea, depression, and tachycardia. CONCLUSION(S): A combination treatment of prednisone, aspirin, folate, and progesterone is associated with a higher live birth rate compared with no treatment in women with IRM. PMID- 16716322 TI - Efficiency of changing the embryo transfer time from day 3 to day 2 among women with poor ovarian response: a prospective randomized trial. AB - OBJECTIVE: To compare the outcome of day 2 and day 3 embryo transfers in women demonstrating poor ovarian response. DESIGN: Prospective randomized clinical trial. SETTING: Private assisted reproductive technology center. PATIENT(S): Two hundred eighty-one women demonstrating poor ovarian response to controlled ovarian hyperstimulation. INTERVENTION(S): Women who were poor responders were randomly allocated to day 2 or day 3 embryo transfer following oocyte retrieval. MAIN OUTCOME MEASURE(S): Implantation rates and pregnancy rates per oocyte retrieval and embryo transfer. RESULT(S): The clinical pregnancy rates per oocyte retrieval (37.2% vs. 21.4%, respectively; P<.05) and per embryo transfer (38.9% vs. 24.1%, respectively; P<.05) were significantly higher in the day 2 embryo transfer group compared with day 3. On the other hand, implantation rates were not different between groups (23.9% vs. 17.2%, respectively; P=.08). CONCLUSION(S): Our results demonstrated that transfering embryos on day 2 could provide an alternative to the management of poor responder patients. PMID- 16716323 TI - Effect of the mode of assisted reproductive technology conception on obstetric outcomes for survivors of the vanishing twin syndrome. AB - Survivors of the "vanishing" twin syndrome, which occurred in dichorionic twins only, and singletons that began as singletons, had similar mean gestational duration and birth weights, as well as similar frequencies of maternal and neonatal complications. This similarity persisted when conventional IVF and IVF plus ICSI cases were separately evaluated. PMID- 16716324 TI - Effects of short-term and long-term metformin treatment on menstrual cyclicity in women with polycystic ovary syndrome. AB - The purpose of this retrospective study was to compare the frequency of menstrual cyclicity between two groups of patients with polycystic ovary syndrome: women who were followed while on metformin for 3-6 months and those who were followed for >6 months. The results showed that metformin is highly effective in normalizing menstrual cyclicity in women with polycystic ovary syndrome (the overall response rate was 69%, with 88% of responders achieving normal cyclicity), especially with a treatment duration of 6 months or longer (the response rate was 40% higher for women who were treated with metformin for >6 months vs. 3-6 months, 77% vs. 55%). PMID- 16716325 TI - Treatment with leuprolide acetate and hormonal add-back for up to 10 years in stage IV endometriosis patients with chronic pelvic pain. AB - This pilot study examined the effect of a low-dose E and pulsed progestogen hormone therapy (HT) regimen for add-back during long-term GnRH-agonist therapy on bone mineral density (BMD) in five patients with stage IV endometriosis. Bone mineral density was stable after initiation of HT for the entire follow-up period (up to 10 years). One patient stopped her treatment on two occasions to conceive and was successful each time with delivery of a normal baby. No patient had return of pelvic pain after HT add-back. PMID- 16716326 TI - Pubertal exposure to estrogenic chemicals affects behavior in juvenile and adult male rats. AB - In this paper, we tested the hypothesis that exposure to estrogens of different source and estrogenic potency at early puberty could affect the development of socio-sexual behavior in the male rat. Puberty is regarded as a second stage of the ontogenetic period, in the sexual maturation of mammals, particularly sensitive to gonadal hormone milieu. We treated animals orally, from postnatal day 23 to 30, with an environmentally compatible dose of bisphenol A (BPA, 40 microg/kg/day) and with a dosage of ethinylestradiol (EE, 0.4 microg/kg/day) comparable to the human oral contraceptives. Exposure to EE altered the temporal pattern of male sexual activity, reducing performance, in the adult animals; slight modifications, in the same direction, were observed with BPA. Short-term behavioral effects were observed in the treated animals, both with BPA and EE: the exploratory drive, directed to a stimulus object and to the environment, as well as to conspecifics, was reduced in the juveniles. Modifications in the circulating T levels were observed after treatments: T was reduced in the juveniles, both with BPA and EE. The decrement persisted in the adult animals but reached significance only in the BPA group. On the whole, effects of pubertal exposure on behavior are more marked with EE than BPA. This can be due to the much higher estrogenic potency of EE; the direction of the behavioral effects of BPA, compared with EE, is however indicative of an estrogenic mechanism. PMID- 16716327 TI - Children's nurses' post-operative pain management practices: an observational study. AB - BACKGROUND: Children continue to experience unrelieved moderate to severe pain post-operatively despite the evidence to guide practice being readily available. Previous studies have relied on self-report measures; there is a need to establish exactly how nurses manage children's pain in practice. OBJECTIVES: To ascertain how nurses actually manage post-operative pain in children and whether pain management practices adhere to current best practice guidelines. DESIGN: An observational study was carried out. Structured and unstructured data were collected. SETTING: A children's surgical ward in the English Midlands caring for children from birth to 16 years. PARTICIPANTS: Registered nurses (n=13) took part in the study. METHODS: Each participant was observed continuously for a period of 5 hours per shift for two to four shifts each. The role of the observer as participant was adopted whereby the researcher could shadow the nurse and act primarily as an observer. Data were collected for 36 shifts (185 hours). RESULTS: While nurses administered analgesic drugs when a child complained of pain, in most other areas practices did not conform to current recommendations and are in need of improvement. Nurses did not, for example, routinely assess a child's pain, nor use non-drug methods of pain relief on a regular basis. CONCLUSIONS: The sub-optimal pain management practices may be attributable to several factors. The professional culture of nursing and/or ward culture may result in poor pain management practices being perpetuated. Nurses may not have the requisite theoretical knowledge to manage pain effectively. A lack of priority may also be attributed to pain management. These areas need exploring further. PMID- 16716328 TI - Higher serum uric acid is associated with increased arterial stiffness in Japanese individuals. AB - Hyperuricemia is postulated to be a risk factor for atherosclerotic diseases, although whether it is independent of classical atherogenic risk factors is controversial. The automatic computer-assisted measurement of brachial-ankle pulse wave velocity (baPWV) is a valid and reproducible method by which to assess arterial stiffness, a potential surrogate marker of early atherosclerosis. By analyzing cross-sectional data from 982 individuals who underwent health screening, we have investigated whether serum uric acid is associated with high baPWV, which was determined as the highest quartile of baPWV values, in a sex specific manner. Multivariate analysis showed that the odds ratios (95% CI) of the highest baPWV quartile across the sex-specific quartiles of serum uric acid were 1.0, 2.80 (0.93-8.40), 2.13 (0.74-6.19), and 2.76 (1.01-7.55) in women, and 1.0, 1.10 (0.55-2.20), 1.97 (1.04-3.75), and 2.24 (1.10-4.56) in men after adjusting for age, total and HDL-cholesterol, BMI, systolic blood pressure, triglycerides, fasting glucose and smoking status. The association between uric acid and high baPWV was observed in both subjects with metabolic syndrome and those without. These data suggest that in both genders, serum uric acid level is associated with increased baPWV, a marker of arterial stiffness, and is in part independent of other conventional risk factors for atherosclerosis and metabolic syndrome. PMID- 16716329 TI - Dispersive liquid-liquid microextraction combined with gas chromatography-flame photometric detection. Very simple, rapid and sensitive method for the determination of organophosphorus pesticides in water. AB - A new method was used for the extraction of organophosphorus pesticides (OPPs) from water samples: dispersive liquid-liquid microextraction (DLLME) coupled with gas chromatography-flame photometric detection (GC-FPD). In this extraction method, a mixture of 12.0 microL chlorobenzene (extraction solvent) and 1.00 mL acetone (disperser solvent) is rapidly injected into the 5.00 mL water sample by syringe. Thereby, a cloudy solution is formed. In fact, the cloudy state is because of the formation of fine droplets of chlorobenzene, which has been dispersed among the sample solution. In this step, the OPPs in water sample are extracted into the fine droplets of chlorobenzene. After centrifuging (2 min at 5000 rpm), the fine droplets of chlorobenzene are sedimented in the bottom of the conical test tube (5.0+/-0.3 microL). Sedimented phase (0.50 microl) is injected into the GC for separation and determination of OPPs. Some important parameters, such as kind of extraction and disperser solvent and volume of them, extraction time, temperature and salt effect were investigated. Under the optimum conditions, the enrichment factors and extraction recoveries were high and ranged between 789-1070 and 78.9-107%, respectively. The linear range was wide (10 100,000 pg/mL, four orders of magnitude) and limit of detections were very low and were between 3 to 20 pg/mL for most of the analytes. The relative standard deviations (RSDs) for 2.00 microg/L of OPPs in water with internal standard were in the range of 1.2-5.6% (n=5) and without internal standard were in the range of 4.6-6.5%. The relative recoveries of OPPs from river, well and farm water at spiking levels of 50, 500 and 5000 pg/mL were 84-125, 88-123 and 93-118%, respectively. The performance of proposed method was compared with solid-phase microextraction (SPME) and single drop microextraction. DLLME is a very simple and rapid (less than 3 min) method, which requires low volume of sample (5 mL). It also has high enrichment factor and recoveries for extraction of OPPs from water. PMID- 16716330 TI - Recent applications of liquid chromatography-mass spectrometry in forensic science. AB - Recent years have seen the development of powerful technologies that have provided forensic scientists with new analytical capabilities, unimaginable only a few years ago. With liquid chromatography-mass spectrometry (LC-MS) in particular, there has been an explosion in the range of new products available for solving many analytical problems, especially for those applications in which non-volatile, labile and/or high molecular weight compounds are being analysed. The aim of this article is to present an overview of some of the most recent applications of LC-MS (/MS) to forensic analysis. To this end, our survey encompasses the period from 2002 to 2005 and focuses on trace analysis (including chemical warfare agents, explosives and dyes), the use of alternative specimens for monitoring drugs of abuse, systematic toxicological analysis and high throughput analysis. It is not the intention to provide an exhaustive review of the literature but rather to provide the reader with a 'flavour' of the versatility and utility of the technique within the forensic sciences. PMID- 16716331 TI - Determination of protein oxidation by mass spectrometry and method transfer to quality control. AB - Characterization and quantitative analysis of oxidation plays an important role in biopharmaceutical development. This study demonstrates an approach to the assessment of susceptible to oxidation methionine residues in monoclonal antibodies and recombinant proteins. A method for the determination of oxidation levels by peptide mapping with mass spectrometric (MS) detection is described and its advantages compared to the UV detection are presented. Good linearity and reproducibility for determination of oxidation with MS detection are demonstrated (R2 > 0.99; RSDs of 4-9%). Aspects of method transfer to quality control group (QC) are discussed. As well, a quick and easy flow injection/MS method is proposed to substitute for peptide map analysis. Peptide coverage, linearity, reproducibility, robustness, sensitivity and quantitative oxidation results are compared for the flow injection/MS and LC/MS approaches. PMID- 16716332 TI - The role of molecular interaction fields on enantioselective and nonselective separation of chiral sulfoxides. AB - The separation of a series of 23 asymmetric sulfoxides, including the three proton pump inhibitors (PPI) omeprazole, lansoprazole and pantoprazole was investigated by HPLC, under reversed-phase elution with amylose tris(3,5 dimethylphenylcarbamate), amylose tris[(S)-1-phenylethylcarbamate] and amylose tris(3,5-dimethoxyphenylcarbamate) chiral stationary phases, CSP1-3, respectively. The whole set of sulfoxides showed better enantioselectivity and enantioresolution on CSP1. However, the three PPI were enantioseparated only when using CSP1 and CSP3. It was observed an improved enantioselectivity and enantioresolution on CSP3. The mechanisms of retention were evaluated by molecular interaction fields (MIF) generated via GRID force field, which yielded the geometric reasons leading to the scenario outlined. The enantioselective and nonselective interactions are discussed in terms of the reported selectivity. The steric structural outline of the CSP nonselective interaction sites is of major importance to deliver the sulfoxides to the chiral selective sites where the enantioselective interactions take place. PMID- 16716333 TI - Determination of oleamide and erucamide in polyethylene films by pressurised fluid extraction and gas chromatography. AB - A pressurized fluid extraction (PFE) and gas chromatography-flame ionization detection (GC-FID) method is proposed to determine the slip agents in polyethylene (PE) films. The study of PFE variables was performed using a fractional factorial design (FFD) for screening and a central composite design (CCD) for optimizing the main variables obtained from the Pareto charts. The variables that were studied include temperature, static time, percentage of cyclohexane and the number of extraction cycles. The final condition selected was pure isopropanol (two times) at 105 degrees C for 16min. The recovery of spiked oleamide and erucamide was around 100%. The repeatability of the method was between 9.6% for oleamide and 8% for erucamide, expressed as relative standard deviation. Finally, the method was applied to determine oleamide and erucamide in several polyethylene films and the results were statistically equal to those obtained by pyrolysis and gas-phase chemiluminescence (CL). PMID- 16716334 TI - Synthesis of ordered large pore SBA-15 spherical particles for adsorption of biomolecules. AB - The synthesis of spherical particles of mesoporous silicates (SBA-15) with mesopore diameter upto 127A, and particle diameter of 4-10 microm has been achieved. The SBA-15 spheres were obtained using pluronics P123 (EO20PO70EO20) as a surfactant coupled with cetyltrimethylammonium bromide (CTAB) as a co surfactant. Ethanol played a very important role in the formation of silica spheres, as it delays the reaction rate of the SBA-15 synthesis. A wide range of pore diameters (28-127 A) of these spherical SBA-15 materials with large surface area >700 m2/g has been synthesized. The effects of temperature, ethanol, CTAB and swelling agent have also been studied. The SBA-15 samples were characterized using small angle X-ray scattering (SAXS), N2 adsorption-desorption, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The spherical SBA-15 after mechanical grinding was used for protein adsorption, and adsorption capacity was compared with that of conventional fibrous SBA-15. The spherical SBA-15 particles with pore diameter of 127 A have a very high capacity of 700 mg/g for lysozyme at pH 7. PMID- 16716335 TI - Continuous flow microextraction combined with high-performance liquid chromatography for the analysis of pesticides in natural waters. AB - Continuous flow microextraction (CFME) combined with high-performance liquid chromatography-ultraviolet (HPLC-UV) detection has been applied to the analysis of five widely used pesticides, simazine, fensulfothion, etridiazole, mepronil and bensulide, present at trace levels in water samples. CFME employs a single organic solvent drop positioned at the tip of a polyether ether ketone (PEEK) tubing, which is immersed in a continuous flowing aqueous sample solution in a 0.5-ml glass chamber. The PEEK tubing acts as the organic drop holder and fluid delivery duct. Analytes are partitioned between the organic drop and the bulk sample solution. Important extraction factors including type of solvent, its volume, sample solution flow rate, extraction time, its pH and addition of salt were investigated. All pesticides exhibit good linearity in the investigated concentration range of 25-250 ng ml(-1) with coefficients of determination (R2) ranging from 0.9879 to 0.9999 under the optimized conditions. Detection limits lower than 4 ng ml(-1) were obtained for all analytes. The method was evaluated by analyzing natural water sample collected from a reservoir in Singapore. This study for the first time demonstrated the compatibility of CFME procedure and HPLC separation. PMID- 16716336 TI - Counter-current chromatography using hexane/surfactant-containing water solvent systems. AB - We developed a n-hexane/surfactant-containing water solvent system in counter current chromatography (CCC) in order to separate hydrophobic compounds. By using the upper phase as the mobile phase, we have separated steroid samples. Retention times of steroids progesterone and delta4-androstene-3,17-dione increased slightly by increasing the concentration below the critical micellar concentration (CMC) of surfactant sodium 1-heptanesulfonate. However, the retention times increased drastically while the SHS concentrations were above the CMC. The partition of these two steroids in the two phases was significantly dependent on the interaction with micelles. Aromatic hydrocarbons were not retained by the lower phase no matter what the surfactant concentrations were. Their hydrophobic interaction with n-hexane greatly exceeded that with the micellar solution. The retention times of esters, however, were only slightly affected by the surfactant addition even above the CMC. The weaker interaction between esters and the micellar solution was probably due to their higher polarity. The micellar solvent systems provide an alternative way for hydrophobic sample separations in CCC, but the performance is limited. PMID- 16716337 TI - Determination of chlorine dioxide in water by gas chromatography-mass spectrometry. AB - A sensitive gas chromatographic method has been established for the determination of chlorine dioxide in water. With weak basic conditions (pH 9.0), chlorine dioxide reacts with iodide to form iodine, which reacts with 2,6-dialkylphenol to form 4-iodo-2,6-dialkylphenol. The volatile organic derivative was extracted with ethyl acetate, and then measured by gas chromatography-mass spectrometry (GC-MS). The reaction of the active proton of 2,6-dialkylphenols (2,6-dimethylphenol, 2,6 di-isopropylphenol and 2,6-di-tert-butylphenol) with iodine was tested, and compared to each other in terms of reactivity and stability of the derivatives. 2,6-dimethylphenol showed rapid reaction with iodine, and its derivative was stable for 2 weeks. The detection limit of chlorine dioxide in water was about 1.0 ng/mL, and the calibration curve showed good linearity with r2 = 0.998. The existent concentration of chlorine dioxide in water was calculated from multiplying the concentration calculated from the calibration curve of 4-iodo-2,6 dimethylphenol by 0.544. The method was sensitive, reproducible and simple enough to permit the reliable analysis of chlorine dioxide at the low ng/mL level in water. PMID- 16716338 TI - Large-scale purification of 13-dehydroxybaccatin III and 10-deacetylpaclitaxel, semi-synthetic precursors of paclitaxel, from cell cultures of Taxus chinensis. AB - The taxane derivatives 13-dehydroxybaccatin III (13-DHB III) and 10 deacetylpaclitaxel (10-DAP) can be used as semi-synthetic precursors of paclitaxel. These precursors were isolated in a simple and economical procedure during purification of paclitaxel from plant cell culture extracts. No additional costs for cell culture or extraction by silica-gel low-pressure chromatography were incurred. Precipitation from dichloromethane/n-hexane followed by HPLC on ODS and silica-gel resins resulted in paclitaxel of 99.5% purity with 80% overall yield. ODS low-pressure chromatography and THF/n-hexane precipitation yielded 13 dehydroxybaccatin III at >99% purity with 87.1% overall yield, and ODS low pressure chromatography alone yielded 10-deacetylpaclitaxel at >90% purity with 93.4% overall yield. These compounds are of sufficient purity for use in semi synthesis of paclitaxel. Both compounds were obtained in high yield at >99.5% purity using ODS-HPLC. The procedures described allow the simultaneous purification of 13-dehydroxybaccatin III, 10-deacetylpaclitaxel, and paclitaxel with only minimal additional expense for reagents or equipment. PMID- 16716339 TI - Molecular recognition of mono- and disaccharides through interaction with p iodophenylboronic acid in capillary electrophoresis with a chemiluminescence detection system. AB - Molecular recognition of mono- and disaccharides was performed making use of the interaction between their diol groups and p-iodophenylboronic acid in capillary electrophoresis (CE) with a chemiluminescence (CL) detection system. p Iodophenylboronic acid acted as an enhancer for luminol-horseradish peroxidase hydrogen peroxide CL reaction. p-Iodophenylboronic acid was injected as a sample into the present system to give a CL peak on the electropherogram. The CL intensities were examined using running buffers including mono- and disaccharides. The CL intensities with 1-methyl-D-glucoside, D-saccharose, D maltose, D-glucose, and D-fructose decreased in this order. The decrease in CL intensity was based on the formation by p-iodophenylboronic acid of cyclic esters with mono- and disaccharides, particularly with those including cis-diol groups. That is, the decrease in CL intensity affected the specific complexation between p-iodophenylboronic acids and saccharides, leading to the molecular recognition of saccharides. We also report separation of a mixture of p-iodophenol and p iodophenylboronic acid as well as estimation of the apparent binding constant between p-iodophenylboronic acid and saccharides taking advantage of their molecular recognition behavior. PMID- 16716340 TI - Octyl-functionalized hybrid silica monolithic column for reversed-phase capillary electrochromatography. AB - Hybrid silica monolithic stationary phase functionalized with octyl groups was synthesized by a two-step acid/base-catalyzed hydrolysis/co-condensation of tetraethoxysilane (TEOS) and n-octyltriethoxysilane (C(8)-TEOS). The influences of determining factors in the sol-gel process such as the monomer ratio and water content on the monolith formation were systematically investigated. An increase in the TEOS/C(8)-TEOS ratio in the polymerization mixture shifted the pore size distribution towards smaller pore diameter with larger pore volume. The optimal TEOS/C(8)-TEOS volume ratio was found to be 90/50, under which condition the median pore diameter of the monolith was around 1.0 microm with pore volume of 3.25 cm(3)/g. The chromatographic characteristics of the monolithic column prepared with the optimized fabrication condition were studied. Some aromatic compounds including alkylbenzenes, polycyclic aromatic hydrocarbons (PAHs) and phenols were successfully separated on the octyl-functionalized silica monolithic column with high column efficiency up to 180,000 plates/m. PMID- 16716341 TI - Extraction and preconcentration of salbutamol and terbutaline from aqueous samples using hollow fiber supported liquid membrane containing anionic carrier. AB - This paper presents a new three-phase liquid-phase microextraction (LPME) strategy for extraction and preconcentration of salbutamol (SB) and terbutaline (TB) from aqueous samples, including urine. The drugs were extracted from 11 ml of aqueous sample (source phase; SP) into an organic phase with microliter volume located inside the pores of a polypropylene hollow fiber, and then back-extracted into 24 microl of a second aqueous solution as the receiving phase (RP), located in the lumen of the hollow fiber. In preliminary experiments, we tried to transport the drugs using a pH gradient between the two sides of the hollow fiber. Due to the existence of both amine and phenolic groups on the drugs, very little transport occurred and enrichment factors (EF) less than one were obtained. Further experiments were done in the presence of bis(2-ethylhexyl) monohydrogenphosphoric acid (D2EHPA) or methyltrioctylammonium chloride (Aliquat 336) in the organic phase, to extract drugs from acidic and basic matrices, respectively. Results showed that transport of drugs from alkaline solution into 1M of sodium bromide occurred when the membrane was impregnated with dihexyl ether containing 20% Aliquat 336. To optimize the EF, the effects of different parameters such as the nature of organic solvent used to impregnate the membrane, compositions and volumes of SP and RP, type and concentration of carrier, extraction time and stirring rate were investigated. Optimal results were obtained in the presence of 0.005 M of NaOH (pH 11.70) in the SP, 1M of NaBr in the RP, 20% of Aliquat 336 in dihexyl ether as membrane impregnation solvent, stirring rate of 500 rpm and extraction time of 60 min. Under these conditions, enrichment factors of 52.9 and 213.1, dynamic linear ranges of 20-5000 and 10 5000, and limits of detection of 2.5 and 0.5 ng/ml were obtained for salbutamol and terbutaline, respectively. Also determination of drugs in environmental water and urine samples in the range of nanograms per millilitre with RSDs<10% was possible using HPLC-photodiode array detection or HPLC-MS. PMID- 16716342 TI - Comparison of complexed species of Eu in alumina-bound and free polyacrylic acid: a spectroscopic study. AB - The speciation of Eu complexed with polyacrylic acid (PAA) and alumina-bound PAA (PAA(ads)) was studied at pH 5 in 0.1 M NaClO(4). Structural parameters were obtained from (7)F(0) -->(5)D(0) excitation spectra measured by laser-induced fluorescence spectroscopy as well as from Eu L(III)-edge extended X-ray absorption fine structure (EXAFS) spectra. The coordination mode was also investigated by infrared spectroscopy. To elucidate the nature of the complexed species, Eu-acetate complexes were used as references. The spectroscopic techniques show that two carboxylate groups with 2-3 (EuPAA) and 4-5 (EuPAA(ads)) water molecules are coordinated to Eu in the first coordination sphere. For EuPAA(ads), the coordination between carboxylate groups and Eu appears to be bidendate. A similar coordination is probable for EuPAA but the EXAFS data indicate a slightly distorted coordination. The results show that the degree of freedom of carboxylate groups is not the same for free or adsorbed PAA. For PAA, the degree of freedom is constrained by the flexibility of the methylene chain. When PAA is adsorbed on alumina, the polymer chains cannot any more be treated as independent chains. One may rather assume formation of aggregates that form an organic layer at the mineral surface presenting a complex arrangement of carboxylate groups. PMID- 16716343 TI - Acidic solution properties of beta-casein-graft-dextran copolymer prepared through Maillard reaction. AB - The copolymer of beta-casein-graft-dextran was prepared using the Maillard reaction and the acidic solution properties of the copolymer were studied with dynamic light scattering. At pH range 4-5 where is close to the isoelectric point of beta-casein, the copolymer forms micelles which are spherical verified by atomic force microscopy imaging. The size and existent time of the micelles depend on the graft degree and the length of dextran side chains of the copolymers. During storage at pH 4.6, the micelles formed by the copolymers with short side chains and low graft degree tend to precipitate, while the micelles formed by the copolymers with long side chains and high graft degree tend to dissociate. The micellization of the copolymers can be suppressed by adding NaCl. Optical microscopy and turbidity studies show that the copolymers dissolved in molecular state and with higher hydrophilicity have better emulsifying ability. PMID- 16716344 TI - Successful simultaneous measurement of cell membrane and cytokine induced phosphorylation pathways [CIPP] in human peripheral blood mononuclear cells. AB - Phenotyping and simple enumeration of peripheral blood mononuclear cells (PBMC) is of limited value for the assessment of many clinical states. As a preferred alternative, cell surface phenotyping may be combined with functional assays for enhanced assessment of altered cells circulating in patients. One simple, yet informative and rapid approach is to examine signaling within individual cells following brief periods of stimulation via flow cytometry. Although monocytes and lymphoid cells can be distinguished based on size, current permeabilization strategies necessary for identifying intracellular phosphorylated signaling molecules largely compromise the labeling of cell surface proteins used to distinguish individual cellular subsets. We have successfully developed conditions that allow for simultaneous detection of cell surface proteins and intracellular phosphorylated proteins in human PBMC following rapid in vitro cytokine stimulation. We analyzed permeabilized CD4, CD8, CD14, CD19, and CD56 expressing cells together with intracellular pSTAT1, pSTAT3, pSTAT5, pSTAT6, pp38 MAPK, or pERK1/2 within total PBMC. Of the permeabilizing conditions tested, 75% methanol enabled superior simultaneous detection of both cell surface and intracellular epitopes. This method enables the rapid functional analysis of subsets within complex cell mixtures and provides an opportunity for assessing abnormalities arising in the setting of acute or chronic inflammatory states. PMID- 16716345 TI - A novel modification of a flow cytometric assay of phosphorylated STAT1 in whole blood lymphocytes for rapid detection of interferon-alpha signal in vivo. AB - Phosphorylation of signal transducer and activator of transcription factor 1 (STAT1) is a key response in the type I interferon (IFN) signal cascade. We developed a novel flow cytometric assay for phosphotyrosine-STAT1 (p-STAT1) to rapidly monitor in vivo IFN signaling. Mouse blood stimulated with mouse IFN alpha was hemolyzed with lysis buffer in place of lymphocyte purification, permeabilized with methanol, and stained with an Alexa Fluor 488-conjugated anti p-STAT1 antibody. The cells were also stained with phycoerythrin (PE)-conjugated anti-CD45 antibody for eliminating debris (CD45-negative) from leukocytes (CD45 positive), and with PE covalently linked to cyanin 5-conjugated anti-Gr-1 antibody for separating lymphocytes (Gr-1-negative) and granulocytes (Gr-1 positive). When whole blood was treated with IFN-alpha, the Alexa Fluor 488 intensity of lymphocytes increased, reaching a peak within 1 h, and this increase was statistically significant at IFN-alpha concentrations of 100 U/mL and higher. When IFN-alpha was administered intravenously to mice, the Alexa Fluor 488 intensity of blood lymphocytes was increased, reaching a peak in 1 h and returning to baseline at 18 h, and this increase was dose-dependent, with statistically significant increases seen at doses of 1,000 U/body and higher. The kinetics and dose-responses of p-STAT1 levels in the spleen, lung, and liver were similar to those in blood lymphocytes. This new flow cytometric assay of p-STAT1 in peripheral blood leukocytes will be useful for examining IFN-alpha signaling and for monitoring tissue response to IFN-alpha in vivo. PMID- 16716346 TI - The effects of the slug biological control agent, Phasmarhabditis hermaphrodita (Nematoda), on non-target aquatic molluscs. AB - The nematode Phasmarhabditis hermaphrodita is used as a commercial biological control agent of slugs in the UK. Although it is known to affect other terrestrial mollusc species, its effects on freshwater molluscs are not known. The present study investigated the effects of P. hermaphrodita on the survival of juvenile Lymnaea stagnalis and Physa fontinalis, two common freshwater snails, at 'spray tank' concentration and a 50% diluted 'spray tank' concentration over a 14 day period. Survival of L. stagnalis was significantly reduced at both application levels but P. fontinalis suffered no mortalities over the experimental period. The possible differential mechanisms of pathology between the two host species are discussed. PMID- 16716347 TI - PARP inhibition prevents postinfarction myocardial remodeling and heart failure via the protein kinase C/glycogen synthase kinase-3beta pathway. AB - The inhibition of glycogen synthase kinase-3beta (GSK-3beta) via phosphorylation by Akt or protein kinase C (PKC), or the activation of mitogen-activated protein kinase (MAPK) cascades can play a pivotal role in left ventricular remodeling following myocardial infarction. Our previous data showed that MAPK and phosphatidylinositol-3-kinase/Akt pathways could be modulated by poly(ADP ribose)polymerase (PARP) inhibition raising the possibility that cardiac hypertrophic signaling responses may be favorably influenced by PARP inhibitors. A novel PARP inhibitor (L-2286) was tested in a rat model of chronic heart failure following isoproterenol-induced myocardial infarction. Subsequently, cardiac hypertrophy and interstitial collagen deposition were assessed; additionally, mitochondrial enzyme activity and the phosphorylation state of GSK 3beta, Akt, PKC and MAPK cascades were monitored. PARP inhibitor (L-2286) treatment significantly reduced the progression of postinfarction heart failure attenuating cardiac hypertrophy and interstitial fibrosis, and preserving the integrity of respiratory complexes. More importantly, L-2286 repressed the hypertrophy-associated increased phosphorylation of panPKC, PKC alpha/betaII, PKC delta and PKC epsilon, which could be responsible for the activation of the antihypertrophic GSK-3beta. This work provides the first evidence that PARP inhibition beneficially modulates the PKC/GSK-3beta intracellular signaling pathway in a rat model of chronic heart failure identifying a novel drug target to treat heart failure. PMID- 16716349 TI - Upregulation of ID protein by growth and differentiation factor 5 (GDF5) through a smad-dependent and MAPK-independent pathway in HUVSMC. AB - GDF5 (growth and differentiation factor five), a member of the TGF-beta superfamily, binds specifically to BMPR1b, BMPR2 and ACTR2a receptors forming a heterodimeric complex, thereby inducing phosphorylation of smad1, 5, 8 and translocation to the nucleus. ID1 (inhibitor of differentiation or DNA binding) is essential for G1 to S phase transition inhibiting DNA binding thereby playing an important role in the control of differentiation, proliferation and angiogenesis. The objective of this study was, therefore, to characterize the signal transduction pathway of GDF5, especially the involvement of ID1, in human umbilical vein smooth muscle cells (HUVSMC). We observed the expression of BMPR1a, BMPR1b, BMPR2, ACTR2a, smad1, smad 5, ID1, ID2 and ID3 in HUVSMC. Application of GDF5 upregulated ID1 and ID3 expression by involvement of the smad signaling pathway. GDF5 caused phorsphorylation of smad1 followed by upregulation of ID1 and ID3. Co-incubation with anti-GDF5 prevented these effects. GDF5 significantly inhibited phosphorylation of p38 MAPK and induced phosphorylation of ERK. The specific inhibitor of p38 MAPK or ERK, SB203580 or U0126 did not induce ID protein expression. Smad1 siRNA transfection inhibited the upregulation of ID protein. GDF5 had chemotactic activity in HUVSMC; this effect was partly blocked by transfection of smad1 or ID1 siRNA. Our results indicate that GDF5 induces ID1 and ID3 in HUVSMC by a smad-dependent, MAPK-independent pathway. GDF5 binds to specific receptors, thereby inducing phosphorylation and translocation of smad1 to the nucleus where it is involved in the regulation of transcription. Since ID1 has been shown to be crucial for cell cycle control, we propose that GDF5 could be involved in the process of angiogenesis. PMID- 16716348 TI - Beta-adrenergic receptors signaling and heart failure in mice, rabbits and humans. PMID- 16716350 TI - Transthyretin: no association between serum levels or gene variants and schizophrenia. AB - It has been proposed that schizophrenia results from an environmental insult in genetically predisposed individuals. Environmental factors capable of modulating transcriptional activity and their carriers could link the genetic and environmental components of schizophrenia. Among these is transthyretin (TTR), a major carrier of thyroid hormones and retinol-binding protein (RBP). Retinoids and thyroid hormones regulate the expression of several genes, both during development and in the adult brain. Decreased TTR levels have been reported in the cerebrospinal fluid of patients with depression and Alzheimer's disease, and the absence of TTR influences behavior in mice. DNA variants capable of altering TTR ability to carry its ligands, either due to reduced transcription of the gene or to structural modifications of the protein, may influence development of the central nervous system and behavior. In the present study we searched for variants in the regulatory and coding regions of the TTR gene, and measured circulating levels of TTR and RBP. We found a novel single nucleotide polymorphism (SNP), ss46566417, 18 bp upstream of exon 4. Neither this SNP nor the previously described rs1800458 were found associated with schizophrenia. In addition, serum TTR and RBP levels did not differ between mentally healthy and schizophrenic individuals. In conclusion, our data does not support an involvement of the TTR gene in the pathophysiology of schizophrenia. PMID- 16716352 TI - Combat veterans show normal discrimination during differential trace eyeblink conditioning, but increased responsivity to the conditioned and unconditioned stimulus. AB - The question addressed in the present study was whether post-traumatic stress disorder (PTSD) results in associative learning impairments. To answer this question, differential trace eyeblink (EB) conditioning was studied in combat veterans with PTSD, combat veterans without PTSD, and non-combat veterans without PTSD. Veterans with PTSD showed normal EB discrimination, suggesting that associative learning is not impaired by PTSD. Veterans with PTSD also showed normal extinction. However, subjects with PTSD showed more EB conditioned responses (CRs), as well as increased CR amplitude. Increased response amplitude to the airpuff unconditioned stimulus presented alone (viz. the unconditioned response), as well as to the airpuff on CS+ trials during conditioning also occurred in the subjects with PTSD. These findings suggest increased reactivity in combat veterans with PTSD, compared to those without PTSD, but such heightened reactivity does not affect somatomotor associative learning. PMID- 16716353 TI - Ionizing radiation to prevent arterial intimal hyperplasia at the edges of the stent: induces necrosis and fibrosis. AB - BACKGROUND: Although ionizing radiation has been proposed for the prevention of intimal hyperplasia in coronary and peripheral arteries in multicenter clinical trials, information is lacking on how irradiation affects arterial histology after stenting and especially how it affects the edges of the stent. We investigated intimal hyperplasia recasting with histological changes in arterial wall at the edges of the stent after arterial stenting followed by adequate external radiation for the prevention of intimal hyperplasia in pigs. MATERIALS AND METHODS: The aorta was experimentally stented in 30 pigs who were then assigned to two groups: irradiation with 20 Gy and a control group with no irradiation. The aorta was resected for morphometric and histological studies 6 weeks after procedure. RESULTS: Intimal thickness was reduced and the intima/media ratio was significantly lower in irradiated groups than in control pigs. In the irradiated group histological examination at the edges of the stent showed thin neointimal proliferation with an intact endothelium. In all sections analyzed in the 20-Gy irradiated group the vascular media at 45 days contained necrotic areas and fibrosis with calcifications. CONCLUSIONS: After arterial injury, adequate ionizing radiation effectively reduces neointimal thickening. Irradiation-induced histological changes include previously undetected recasting with necrosis and fibrosis at the arterial edges of the stent. The parietal recasting we observed in animal arteries irradiated at high doses is unclear and a cause of concern especially after clinical spontaneous dissection was recently reported. The use of ionizing radiation for the prevention of arterial restenosis awaits confirmation with a long-term follow-up including specific experimental histological analyses. PMID- 16716351 TI - Clinical vs. self-report versions of the quick inventory of depressive symptomatology in a public sector sample. AB - OBJECTIVES: Recent work using classical test theory (CTT) and item response theory (IRT) has found that the self-report (QIDS-SR(16)) and clinician-rated (QIDS-C(16)) versions of the 16-item quick inventory of depressive symptomatology were generally comparable in outpatients with nonpsychotic major depressive disorder (MDD). This report extends this comparison to a less well-educated, more treatment-resistant sample that included more ethnic/racial minorities using IRT and selected classical test analyses. METHODS: The QIDS-SR(16) and QIDS-C(16) were obtained in a sample of 441 outpatients with nonpsychotic MDD seen in the public sector in the Texas Medication Algorithm Project (TMAP). The Samejima graded response IRT model was used to compare the QIDS-SR(16) and QIDS-C(16). RESULTS: The nine symptom domains in the QIDS-SR(16) and QIDS-C(16) related well to overall depression. The slopes of the item response functions, a, which index the strength of relationship between overall depression and each symptom, were extremely similar with the two measures. Likewise, the CTT and IRT indices of symptom frequency (item means and locations of the item response functions, b(i) were also similar with these two measures. For example, sad mood and difficulty with concentration/decision making were highly related to the overall depression severity with both the QIDS-C(16) and QIDS-SR(16). Likewise, sleeping difficulties were commonly reported, even though they were not as strongly related to overall magnitude of depression. CONCLUSION: In this less educated, socially disadvantaged sample, differences between the QIDS-C(16) and QIDS-SR(16) were minor. The QIDS-SR(16) is a satisfactory substitute for the more time consuming QIDS-C(16) in a broad range of adult, nonpsychotic, depressed outpatients. PMID- 16716354 TI - Recipient blood pre-transplant transfusion prolongs hepatic allograft survival in rats. AB - BACKGROUND: The pre-transplant administration of donor antigens to recipients is reported to prolong transplanted organ survival. We investigated the effect of pre-transplant intraportal administration of recipient blood on rat hepatic allograft survival. MATERIALS AND METHODS: Male LEW (RT1l) and ACI (RT1a) rats were used as transplant recipients and donors, respectively. Before transplantation, donors were transfused with recipient blood. Experimental animals were divided into groups as follows: group I, no treatment; group II, pre treatment with recipient blood via the penile vein 7 days before transplantation; group III, pre-treatment with recipient blood via the portal vein 5 days before transplantation; and group IV, pre-treatment with recipient blood via the portal vein 7 days before transplantation. Serum interferon (IFN)-gamma concentrations were measured post-operatively. RESULTS: Animals in group I survived a mean of 10.1 +/- 0.7 days. The survival of groups II and III was 10.6 +/- 1.6 and 13.1 +/ 0.9 days, respectively. The survival rate in group IV was prolonged significantly to 33.7 +/- 2.6 days. Serum concentrations of IFN-gamma were increased significantly in group IV, as compared with group I. The ratio of OX76+CD4+ or OX76+CD8+ T cells to OX76-CD4+ or OX76-CD8+ T cells was greater in group IV, as compared group I. OX76+CD8+ T cells from hepatic allografts in group IV expressed IFN-gamma and interleukin (IL)-10, but not IL-2 mRNA. Apoptotic hepatic infiltrates were greater in group IV, as compared to group I. CONCLUSION: The cytokine profile of donor CD8+ T cells from allografts treated by the intraportal administration of recipient blood is associated with apoptosis of graft-infiltrating cells and the prolonged survival of hepatic allografts in rats. PMID- 16716355 TI - Systolic contraction within aneurysmal rabbit myocardium following transplantation of autologous skeletal myoblasts. AB - OBJECTIVE: Transplantation of autologous skeletal myoblasts (SKMB) into infarcted heart (or cellular cardiomyoplasty, CCM) augments myocardial performance in animal models of myocardial infarction. However, the effect of CCM in the setting of ventricular aneurysm has not been evaluated. This study analyzes the effects of transplanted SKMB on regional wall motion in a rabbit model of postinfarct ventricular aneurysm. We hypothesize that CCM, performed early after myocardial infarction, prevents the progression of dyskinetic wall motion. METHODS: Twenty six rabbits underwent apical left ventricular cryoinfarction and soleus muscle biopsy for in vitro isolation of skeletal myoblasts. At 2 weeks postinfarct, the presence of ventricular aneurysm was detected in 23/26 animals by sonomicrometry and micromanometry. Seventeen of 23 animals were randomized to receive either 108 autologous myoblasts (CCM) or vehicle. Regional stroke work, percent systolic shortening, and synchronicity of regional wall motion were determined prior to, and four weeks following, injection (CCM; n = 9; vehicle, n = 8). Wall motion was evaluated under baseline and stress (dobutamine, 10 (g/kg/min) conditions. Six animals did not undergo randomization, but their hearts were used to measure the size of infarction. RESULTS: Four weeks following treatment of animals with ventricular aneurysm, systolic contractile activity was present in most animals treated with myoblasts but in none treated with vehicle (5/7 versus 0/6, respectively, P < 0.05). Dobutamine tended to accentuate the differences seen at baseline between the groups. CONCLUSIONS: This study demonstrates a high incidence of systolic contractile activity in a previously aneurysmal region of myocardium following CCM and may represent a novel therapy for the prevention and treatment of postinfarct aneurysm. PMID- 16716356 TI - The flow phase of wound metabolism is characterized by stimulated protein synthesis rather than cell proliferation. AB - BACKGROUND: Healing of a skin wound requires net protein deposition to repair the tissue defect and new epidermal cells to cover the wound. However, the true course of changes in rates of cell proliferation and protein deposition following skin injury has not been previously determined. This experiment was to measure DNA fractional synthetic rate (FSR), reflecting cell division, and protein FSR and fractional breakdown rate (FBR) in skin wound at three times after injury. MATERIALS AND METHODS: The experiment consisted of a surgery and a tracer infusion on separate days. During the surgery (day 0), a donor wound was created and indwelling catheters were inserted into the carotid artery and jugular vein under general anesthesia. On day 1, day 3, or day 7, stable isotope tracer infusion was performed in conscious rabbits to determine DNA FSR and protein FSR and FBR in the wound. RESULTS: Protein FSR and FBR in the day 7 wound were 20.5 +/- 8.4 and 12.6 +/- 4.7%/day, respectively, which were greater (P < 0.01-0.05) than the corresponding values in the day 1 and day 3 wounds. Net protein deposition (FSR-FBR) in day 7 wound (7.9 +/- 6.0%/day) was greater (P < 0.05) than in day 3 wound (0.8 +/- 2.4%/day). DNA FSRs were 1.94 +/- 0.58, 2.43 +/- 0.96, 2.86 +/- 0.90%/day in the day 1, day 3 and day 7 wounds, respectively (P = 0.2). CONCLUSIONS: The flow phase in the wound is characterized by increased protein synthesis rather than cell proliferation; net protein deposition in the wound is a major cause of protein requirements in severe burns. PMID- 16716357 TI - Evaluation of hepatic interleukin-6 secretion following portal vein ligation using a minimal surgical stress model. AB - BACKGROUND: Animal portal vein ligation (PVL) model requires laparotomy under general anesthesia, which induces systemic inflammatory response, and makes the analysis of early signals for hepatic regeneration difficult. We developed a nonstress PVL model, which performs the ligation after surgical stress is subsided, and evaluated the hepatic IL-6 production at early phase of PVL. MATERIALS AND METHODS: After laparotomy under general anesthesia, a monofilament suture was placed around the left portal vein and knotted, but not tied, and the two ends were delivered through each flank. Seven days after laparotomy, PVL was created by pulling the two ends of the ligature, tightening the ligature. RESULTS: Biochemical assays indicated that the surgical stress of laparotomy resolved on day 7. Rate of liver regeneration of the nonligated lobe in this model was similar to that in the conventional PVL model. The serum interleukin-6 concentration rose rapidly (within 3 h) after PVL and remained elevated until 6 h. RT-PCR demonstrated that transcripts of IL-6 gene significantly increased 3 h after ligation in the nonembolized lobes. Furthermore, in situ hybridization for IL-6 mRNA was stained at sinusoidal endothelial cells, especially periportal area. CONCLUSIONS: Our PVL model minimizing surgical stress was comparable to the stress of clinical portal vein embolization. Using this model, we found an increased expression of IL-6 mRNA in the sinusoidal endothelial cells predominantly at the periportal area. PMID- 16716358 TI - Early increased MT1-MMP expression and late MMP-2 and MMP-9 activity during Angiotensin II induced aneurysm formation. AB - BACKGROUND: Angiotensin II (Ang II) is associated with a variety of cardiovascular diseases including aneurysm formation. The aim of this study was to evaluate the temporal changes in MT1-matrix metalloproteinase (MMP) and MMP-2 and -9 expression and activity during the course of Ang II induced experimental aneurysm formation. METHODS: Apolipoprotein E knockout mice (ApoE null) were infused with either 1000 ng/kg/min of Ang II (n = 20) or saline (n = 20) and then sacrificed at 7, 14, 21, and 28 days of infusion (n = 5/group/strain). Aortic diameters were measured by digital microscopy. Systolic blood pressure (SBP) was measured in the rodent tail. Suprarenal abdominal aortas had MT1-MMP mRNA levels and MMP-2 and MMP-9 mRNA levels and activity quantitated using reverse transcriptase-polymerase chain reaction (rt-PCR) and gelatin zymography, respectively. Statistical analyses included nonpaired t-test, Fisher's exact test, and analysis of variance (ANOVA). RESULTS: Aneurysms occurred in 40, 40, 20, and 80% of ApoE null-Ang II mice at 7, 14, 21, and 28 days, respectively. An early and significant rise in MT1-MMP mRNA occurred in ApoE null mice infused with Ang II mice, while there was no significant change in MMP-2 or MMP-9 mRNA levels. Total MMP-2 and MMP-9 activity increased over time in ApoE null mice infused with Ang II, peaking at 28 days (ANOVA, P < 0.01). SBP was significantly elevated by 7 days in ApoE null mice infused with Ang II compared to ApoE null mice infused with saline (123 +/- 16 versus 102 +/- 6 mm Hg, P < 0.05). CONCLUSIONS: Angiotensin II induces an early increase in aortic MT1-MMP expression with a subsequent increase in MMP-2 and MMP-9 activity. The process by which these changes cause aneurysm formation warrants further investigation. PMID- 16716359 TI - Early cognitive impairment predicts long-term depressive symptoms and quality of life after stroke. AB - OBJECTIVE: The aim of the present study was to examine the predictive value of cognitive impairment in the acute phase after stroke as a risk factor for long term (six to ten months after stroke) depressive symptoms (DS) and a reduced quality of life (QOL), independent of demographic and neurological predictors. METHODS: We evaluated 143 patients within the first 3 weeks post-stroke. Predictor variables included domain-specific cognitive function, demographic data, vascular risk factors, lesion characteristics, and clinical factors. Predictor variables associated with long-term DS (Montgomery Asberg Depression Rating Scale >or=7) and QOL (Stroke-Specific Quality of Life Scale) were identified with multiple logistic and linear regression. RESULTS: Long-term DS were independently predicted by cognitive impairment at baseline, DS at baseline, female sex, diabetes mellitus, and previous TIA(s). Cognitive impairment, increasing age, and functional dependence predicted a reduced QOL, whereas hypercholesterolaemia predicted a better QOL. Among all cognitive disorders, unilateral neglect was the greatest risk factor for DS after 6 months, whereas a disorder in visual perception and construction affected QOL the most. CONCLUSIONS: Cognitive impairment and vascular risk factors are important predictors of long-term DS and QOL after stroke. The prognostic value of cognition suggests a reactive component in the development or continuation of long-term DS. PMID- 16716360 TI - Overexpression of uncoupling protein 4 promotes proliferation and inhibits apoptosis and differentiation of preadipocytes. AB - Uncoupling proteins are a family of mitochondrial proteins involved in energy metabolism. We previously showed that uncoupling protein 4 (UCP4) is differentially expressed in omental adipose tissue in diet-induced obese and normal rats. However, the effect of UCP4 on adipocytes is unclear. In this work, we established a stable preadipocyte cell line overexpressing UCP4 to observe the direct effect of UCP4 on adipocytes. Cells overexpressing UCP4 showed significantly attenuated differentiation of preadipocytes into adipocytes. During differentiation, expression of adipogenesis-associated markers such as fatty acid synthetase, peroxisome proliferator-activated receptor gamma, CCAAT enhancer binding protein alpha, adipocyte lipid binding protein and lipoprotein lipase were downregulated. Preadipoctes expressing UCP4 grew faster and more of them stayed in S phase compared to control cells. In addition, UCP4 overexpression protected preadipocytes from apoptosis induced by serum deprivation. Our results demonstrate that overexpression of UCP4 can promote proliferation and inhibit apoptosis and differentiation of preadipocytes. PMID- 16716361 TI - Ouabain-induced hypertension enhances left ventricular contractility in rats. AB - Chronic ouabain treatment produces hypertension acting on the central nervous system and at vascular levels. However, cardiac effects in this model of hypertension are still poorly understood. Hence, the effects of hypertension induced by chronic ouabain administration ( approximately 8 microg day(-1), s.c.) for 5 weeks on the cardiac function were studied in Wistar rats. Ouabain induces hypertension but not myocardial hypertrophy. Awake ouabain-treated rats present an increment of the left ventricular systolic pressure and of the maximum positive and negative dP/dt. Isolated papillary muscles from ouabain-treated rats present an increment in isometric force, and this effect was present even when inotropic interventions (external Ca(2+) increment and increased heart rate) were performed. However, the sarcoplasmic reticulum activity and the SERCA-2 protein expression did not change. On the other hand, the activity of myosin ATPase increased without changes in myosin heavy chain protein expression. In addition, the expression of alpha(1) and alpha(2) isoforms of Na(+), K(+)-ATPase also increased in the left ventricle from ouabain-hypertensive rats. The present results showed positive inotropic and lusitropic effects in hearts from awake ouabain-treated rats, which are associated with an increment of the isometric force development and of the activity of myosin ATPase and expression of catalytic subunits of the Na(+), K(+)-ATPase. PMID- 16716362 TI - ENOS is not activated by nebivolol in human failing myocardium. AB - Nebivolol is a highly selective beta(1)-adrenoceptor blocker with additional vasodilatory properties, which may be due to an endothelial-dependent beta(3) adrenergic activation of the endothelial nitric oxide synthase (eNOS). beta(3) adrenergic eNOS activation has been described in human myocardium and is increased in human heart failure. Therefore, this study investigated whether nebivolol may induce an eNOS activation in cardiac tissue. Immunohistochemical stainings were performed using specific antibodies against eNOS translocation and eNOS serine(1177) phosphorylation in rat isolated cardiomyocytes, human right atrial tissue (coronary bypass-operation), left ventricular non-failing (donor hearts) and failing myocardium after application of the beta-adrenoceptor blockers nebivolol, metoprolol and carvedilol, as well as after application of BRL 37344, a specific beta(3)-adrenoceptor agonist. BRL 37344 (10 microM) significantly increased eNOS activity in all investigated tissues (either via translocation or phosphorylation or both). None of the beta-blockers (each 10 microM), including nebivolol, increased either translocation or phosphorylation in any of the investigated tissues. In human failing myocardium, nebivolol (10 microM) decreased eNOS activity. In conclusion, nebivolol shows a tissue-specific eNOS activation. Nebivolol does not activate the endothelial eNOS in end-stage human heart failure and may thus reduce inhibitory effects of NO on myocardial contractility and on oxidative stress formation. This mode of action may be of advantage when treating heart failure patients. PMID- 16716364 TI - Effect of resistance exercise on dehydroepiandrosterone sulfate concentrations during a 72-h recovery: relation to glucose tolerance and insulin response. AB - Serum dehydroepiandrosterone sulfate (DHEA-S) concentration is known to be associated with the whole-body insulin sensitivity. The main purpose of the study was to investigate the effect of resistance exercise on DHEA-S concentration during a 72 h post-exercise recovery, and its relation to glucose tolerance and insulin sensitivity. Morning fasted serum samples was obtained from 19 male volunteers (aged 21.1+/-0.4 years) 24 h before the onset of exercise and 24 h, 48 h, and 72 h following exercise for measurements of DHEA-S, cortisol, and TNF alpha. Oral glucose tolerance test (OGTT) and insulin response were determined 24 h before and 48 h after exercise. We found that resistance exercise causes a delayed suppression in serum DHEA-S levels during recovery (48 h and 72 h). This exercise challenge did not affect glucose tolerance, but insulin response during OGTT was significantly elevated. The increased insulin level was not associated with serum levels of cortisol and TNF-alpha. In conclusion, the present study found that resistance exercise has a DHEA-S lowering effect that persisted for 72 h. This change could be related to the elevated insulin concentrations during OGTT. PMID- 16716363 TI - Evaluation of antioxidant activity and potential toxicity of 1-buthyltelurenyl-2 methylthioheptene. AB - The aim of the present study was to evaluate pharmacological and toxicological properties of 1-buthyltelurenyl-2-methylthioheptene (compound 1). In vitro, compound 1 at 1 microM was effective in reducing lipid peroxidation induced by Fe/EDTA. Compound 1 presented neither thiol peroxidase nor thiol oxidase activity and did not change delta-ALA-D (delta-aminolevulinate dehydratase) activity (10 400 microM). Calculated LD(50) of compound 1, administered by oral route, was 65.1 micromol/kg. Rats treated with compound 1 did not reveal any motor impairment in the open field. Hepatic, renal and cerebral lipid peroxidation in treated rats did not differ from those in control rats. Conversely, 0.5 micromol/kg of compound 1 decreased lipid peroxidation in spleen. Delta-ALA-D activity in liver and spleen was inhibited in rats treated with the higher dose of compound 1 but no significant differences were detected in renal delta-ALA-D activity. AST (aspartate aminotransferase) and ALT (alanine aminotransferase) activities as well as urea and creatinine levels were increased by high doses of compound 1 (50-75 micromol/kg). Compound 1 induced a significant decrease in plasma triglyceride levels but none of the doses tested changed the cholesterol level. This is a promising compound for more detailed pharmacological studies involving organotellurium compounds. PMID- 16716365 TI - Statistical tests for analysing directed movement of self-organising animal groups. AB - We discuss some theory concerning directional data and introduce a suite of statistical tools that researchers interested in the directional movement of animal groups can use to analyse results from their models. We illustrate these tools by analysing the results of a model of groups moving under the duress of certain informed indistinguishable individuals, that arises in the context of honeybee (Apis mellifera) swarming behaviour. We modify an existing model of collective motion, based on inter-individual social interactions, allowing knowledgeable individuals to guide group members to the goal by travelling through the group in a direct line aligned with the goal direction. PMID- 16716366 TI - Non-random distribution of instability-associated chromosomal rearrangement breakpoints in human lymphoblastoid cells. AB - Genomic instability is observed in tumors and in a large fraction of the progeny surviving irradiation. One of the best-characterized phenotypic manifestations of genomic instability is delayed chromosome aberrations. Our working hypothesis for the current study was that if genomic instability is in part attributable to cis mechanisms, we should observe a non-random distribution of chromosomes or sites involved in instability-associated rearrangements, regardless of radiation quality, dose, or trans factor expression. We report here the karyotypic examination of 296 instability-associated chromosomal rearrangement breaksites (IACRB) from 118 unstable TK6 human B lymphoblast, and isogenic derivative, clones. When we tested whether IACRB were distributed across the chromosomes based on target size, a significant non-random distribution was evident (p<0.00001), and three IACRB hotspots (chromosomes 11, 12, and 22) and one IACRB coldspot (chromosome 2) were identified. Statistical analysis at the chromosomal band-level identified four IACRB hotspots accounting for 20% of all instability associated breaks, two of which account for over 14% of all IACRB. Further, analysis of independent clones provided evidence within 14 individual clones of IACRB clustering at the chromosomal band level, suggesting a predisposition for further breaks after an initial break at some chromosomal bands. All of these events, independently, or when taken together, were highly unlikely to have occurred by chance (p<0.000001). These IACRB band-level cluster hotspots were observed independent of radiation quality, dose, or cellular p53 status. The non random distribution of instability-associated chromosomal rearrangements described here significantly differs from the distribution that was observed in a first-division post-irradiation metaphase analysis (p=0.0004). Taken together, these results suggest that genomic instability may be in part driven by chromosomal cis mechanisms. PMID- 16716367 TI - Balance marks cognitive changes in old age because it reflects global brain atrophy and cerebro-arterial blood-flow. AB - In healthy old age biomarkers such as Balance robustly correlate with measures of mental abilities such as scores on tests of intelligence, reaction times and memory. A plausible explanation is that balance reflects general physiological fitness and so also neurophysiological integrity, but direct evidence is lacking. Brain scans measured age-associated loss of brain volume and cerebro-arterial blood flow (CBf) in 69 volunteers aged from 62 to 81 years who also took the Tinetti Balance test battery, 3 tests of fluid intelligence, 3 tests of decision speed and a memory test. Balance, but not atrophy or CBf, predicted intelligence test scores. Balance, atrophy, and CBf all independently predicted speed and memory scores but, after variance in atrophy and CBf had been considered, predictions from Balance were no longer significant. It appears that in these tests Balance marks cognitive performance in old age because it reflects gross age-related neurophysiological changes. PMID- 16716368 TI - Clerodanes and other constituents of Cleidion spiciflorum. AB - The polyoxygenated clerodane, spiciflorin (1a), was isolated from Cleidion spiciflorum (Burm. f.) Merr. (Euphorbiaceae). Other constituents were the glucoside of anol (2), columbin, scopoletin, 3,3',4-O-trimethylellagic acid, acetylaleuritolic acid, common triterpenes and phenols. PMID- 16716369 TI - Location and biosynthesis of monoterpenyl fatty acyl esters in rose petals. AB - The upper epidermal layer of cells and the epicuticular wax surface of Lady Seton rose petals are sites of biosynthesis and accumulation, respectively, of a family of terpenyl fatty acyl esters. These esters are based mainly on the acyclic monoterpene alcohol geraniol coupled primarily to fatty acids of chain lengths 16 20 and in mass terms represent from 14% to 64% of the total monoterpenes present in the petals. The lipophilic nature of these non-volatile esters of the monoterpene alcohols contrasts with that of the lipophilic volatile parent alcohols themselves and with the hydrophilic, non-volatile, glucoside derivative of the other principal petal fragrant compounds, the phenylpropanoids, beta phenyl ethanol and benzyl alcohol. These latter compounds are also synthesised and are resident in the petal. Biosynthetic studies confirmed that the petal upper epidermal cell layer has the capacity to incorporate mevalonic acid into the monoterpene component of the fatty acyl ester. The biosynthesis of the monoterpene component of the fatty acyl ester occurs via the mevalonic acid pathway in Lady Seton as well as in the hybrid tea rose Fragrant Cloud. In the latter flower the biosynthesis of geraniol was biosynthetically trans as was the formation of nerol and citronellol. Both geraniol and nerol were shown to be precursors of citronellol via an NADPH dependent reductase reaction. Oleic acid is assimilated into the acyl moiety of the terpenyl ester in Lady Seton isolated petal discs. It is probable that the lipophilic non-volatile terpenyl fatty acyl esters represent a stable storage form of the corresponding alcohols from their residency within the epicuticular wax layer. These acyl esters may realise, on hydrolysis, additional aroma notes from the living flower and potentially commercially significant quantities of the fragrant terpenols during oil of rose essence production. PMID- 16716370 TI - A galloylated cyanogenic glycoside from the Australian endemic rainforest tree Elaeocarpus sericopetalus (Elaeocarpaceae). AB - A cyanogenic glycoside - 6'-O-galloylsambunigrin - has been isolated from the foliage of the Australian tropical rainforest tree species Elaeocarpus sericopetalus F. Muell. (Elaeocarpaceae). This is the first formal characterisation of a cyanogenic constituent in the Elaeocarpaceae family, and only the second in the order Malvales. 6'-O-galloylsambunigrin was identified as the principal glycoside, accounting for 91% of total cyanogen in a leaf methanol extract. Preliminary analyses indicated that the remaining cyanogen content may comprise small quantities of sambunigrin, as well as di- and tri-gallates of sambunigrin. E. sericopetalus was found to have foliar concentrations of cyanogenic glycosides among the highest reported for tree leaves, up to 5.2 mg CN g(-1) dry wt. PMID- 16716371 TI - Evaluation of a standard immunofluorescence assay and a new flow cytometric method for the detection of autoreactive antibodies in dogs with tumours. AB - The major purpose of the presented study was to develop and to evaluate a flow cytometry-based assay (IIFC) for the determination of autoreactive antibodies in sera from canine cancer patients. A blinded study demonstrated the poor reproducibility of the standard, slide-based and microscopically evaluated indirect immunofluorescence test (IIF), especially with sera displaying a cytoplasmic reactivity. In the IIFC, the intra assay coefficient of variance ranged between 5% and 11%, the inter assay variance between 8% and 25%. The IIFC resulted in significantly less positive results among canine cancer patients (16%) than the IIF (40%). The latter results were due to low titered sera indicating that the standard assay may lead to a high proportion of false positive results. The limitation of the IIFC is that no conclusions can be made about the sub cellular localization of the fluorescence. However, this cytometry based assay makes a more objective and standardized detection of canine autoreactive antibodies possible. PMID- 16716372 TI - Lack of activity of cadmium in in vitro estrogenicity assays. AB - Prompted by reports about strong estrogenic effects of cadmium, attempts were made to reproduce these observations using the yeast estrogen screen (YES) and the E-Screen assays. For the first time, possible activation of the Src/MAPK pathway was also investigated. In the YES, only a slight activation (10% of a maximal effect) of the estrogen receptor alpha (ERalpha) was observed at cadmium concentrations between 5 x 10(-7) M and 5 x 10(-6) M. In the E-Screen assay, carried out by two laboratories, the heavy metal was without observable cell proliferative effects when tested in the range between 6 x 10(-11) M and 1 x 10( 5) M. However, in both assays, cadmium led to a reduction of the effects of 17beta-estradiol (E2). Treatment of MCF-7 human breast cancer cells with 1 x 10( 7) M cadmium failed to induce phosphorylation of Src and the MAP kinases Erk1 and Erk2-effects shown to occur with E2 and epidermal growth factor (EGF). In summary, we were unable to confirm the strong estrogenicity of cadmium reported recently by a number of laboratories. This apparent absence of effects in our hands is not due to a lack of uptake of the metal or to effective protection against cadmium by high levels of glutathione or metallothionein, since toxicity and an antagonism of E2 responses were observed both in the YES and the E-Screen. PMID- 16716373 TI - DTX5c, a new OA sulphate ester derivative from cultures of Prorocentrum belizeanum. AB - Prorocentrum belizeanum is a dinoflagellate known for its okadaic acid (OA) and dinophysitoxins (DTXs) production, both OA and DTX are polyether toxins of the Diarrhetic Shellfish Poisoning (DSP) group. We have recently published the isolation of a new diol-ester of okadaic acid from cultures of P. belizeanum. On this occasion we present a new sulphated water-soluble derivative, DTX-5c, isolated from this microalga, whose structure was established on the basis of its spectroscopical data. PMID- 16716374 TI - Genetic analysis of poxvirus mRNA cap methyltransferase: suppression of conditional mutations in the stimulatory D12 subunit by second-site mutations in the catalytic D1 subunit. AB - The guanine-N7 methyltransferase domain of vaccinia virus mRNA capping enzyme, composed of catalytic vD1(498-844) and stimulatory vD12 subunits, can function in vivo in Saccharomyces cerevisiae in lieu of the essential cellular cap methyltransferase Abd1. Coexpression of both poxvirus subunits is required to complement the growth of abd1Delta cells. A double-alanine scan of the vD12 protein identified lethal and temperature-sensitive vD12 alleles. We used this mutant collection to perform a forward genetic screen for compensatory changes in the catalytic subunit that suppressed the growth phenotypes of the vD12 mutants. The screen reiteratively defined a small ensemble of amino acids in vD1(498-844) at which mutations restored methyltransferase function in conjunction with defective vD12 proteins. Reference to the crystal structure of the microsporidian cap methyltransferase suggests that distinct functional classes of suppressors were selected, including: (i) those that map to surface-exposed loops, which likely comprise the physical subunit interface; (ii) those in or near the substrate binding sites, which presumably affect or mimic inter-subunit allostery. PMID- 16716375 TI - Morbillivirus nucleoprotein possesses a novel nuclear localization signal and a CRM1-independent nuclear export signal. AB - Morbilliviruses, which belong to the Mononegavirales, replicate its RNA genome in the cytoplasm of the host cell. However, they also form characteristic intranuclear inclusion bodies, consisting of nucleoprotein (N), in infected cells. To analyze the mechanisms of nucleocytoplasmic transport of N protein, we characterized the nuclear localization (NLS) and nuclear export (NES) signals of canine distemper virus (CDV) N protein by deletion mutation and alanine substitution of the protein. The NLS has a novel leucine/isoleucine-rich motif (TGILISIL) at positions 70-77, whereas the NES is composed of a leucine-rich motif (LLRSLTLF) at positions 4-11. The NLS and NES of the N proteins of other morbilliviruses, that is, measles virus (MV) and rinderpest virus (RPV), were also analyzed. The NLS of CDV-N protein is conserved at the same position in MV-N protein, whereas the NES of MV-N protein is located in the C-terminal region. The NES of RPV-N protein is also located at the same position as CDV-N protein, whereas the NLS motif is present not only at the same locus as CDV-N protein but also at other sites. Interestingly, the nuclear export of all these N proteins appears to proceed via a CRM1-independent pathway. PMID- 16716376 TI - TNFalpha and GM-CSF-induced activation of the CAEV promoter is independent of AP 1. AB - Caprine arthritis encephalitis virus transcription is under the control of the viral promoter within the long terminal repeat. Previous studies with the closely related maedi visna lentivirus have indicated that viral transcription is dependent upon the AP-1 transcription factor. Other studies have indicated a potential role for the cytokines TNFalpha and GM-CSF in CAEV pathogenesis by increasing viral loads in infected tissues. The hypotheses that AP-1 transcription factors are necessary for transcriptional activation of the CAEV promoter and that CAEV transcriptional activation results from treatment with the cytokines GM-CSF and TNFalpha were tested with a stably transduced U937 cell line. Here, we found that TNFalpha and GM-CSF activated CAEV transcription in U937 cells. However, this activation effect was not blocked by SP600125, an inhibitor of Jun N-terminal kinase. SP600125 effectively prevented Jun phosphorylation in cells subsequently treated with cytokines. The cytokines TNFalpha and GM-CSF therefore activate CAEV transcription, and this effect occurs independently of AP-1. A set of progressive deletion mutants was utilized to show that TNFalpha-induced expression depends on an element or elements within the U3 70-bp repeat. PMID- 16716377 TI - Expression of Kaposi's sarcoma-associated herpesvirus-encoded K10/10.1 protein in tissues and its interaction with poly(A)-binding protein. AB - The K10/10.1 protein is encoded by a cluster of interferon regulatory factor (IRF) homologues in the Kaposi's sarcoma-associated herpesvirus (KSHV, human herpesvirus 8, HHV-8) genome. In the present study, we showed that an anti-K10 antibody reacted with a 110-kDa protein encoded by the K10/10.1 gene of KSHV in KSHV-infected primary effusion lymphoma (PEL) cell lines. Expression of K10/10.1 protein was induced by phorbol ester in KSHV-infected cells. A reporter gene assay demonstrated that K10/10.1 protein did not influence promoter activity of human interferon genes, regardless of its homology to human IRFs. Poly(A)-binding protein (PABP) was identified as a partner of K10/10.1 protein. Immunoprecipitation revealed that K10/10.1 protein interacted with PABP specifically in PEL cell lines. IFA revealed co-localization of K10/10.1 protein and PABP in the nucleus of KSHV-infected cells. These data suggest that K10/10.1 protein may affect the translational status or stability of mRNA in host cells. PMID- 16716378 TI - Polychlorinated dibenzo-p-dioxins, dibenzofurans, and dioxin-like polychlorinated biphenyls in rice plants: possible contaminated pathways. AB - Uptake of dioxins was confirmed in rice plants. We determined (i) dioxin concentrations and isomer profiles in leaf+stem samples of rice plants grown in three soils with different dioxin concentrations and isomer profiles and (ii) temporal changes in dioxin concentrations and isomer profiles in rice grown in a paddy field on which agricultural chemicals containing dioxin impurities had been used in previous years. Principal-component and chemical mass-balance analyses of the data showed that dioxin concentrations and isomer profiles in the rice grown in the different soils were similar, even though the dioxin concentrations and isomer profiles of the soils were different. However, the dioxin concentrations in the rice were influenced by dioxin levels in the atmospheric gas phase. Changes in dioxin levels during the growing period of the rice were also strongly influenced by adhesion of the soil to plant components during the early stage of growth, but gradually the influence of the atmospheric gas phase became predominant. Calculated dietary uptake estimates indicated that, even in rice grown in Japan on soil contaminated with high concentrations of dioxins, the amount of dioxins ingested by humans from rice is extremely small, and the amount of dioxins ingested by livestock from forage rice is similar to that for other forage crops. PMID- 16716379 TI - A critical review of patient-rated quality of life studies of long-term survivors of cervical cancer. AB - OBJECTIVES: Increasing survival rates of cervical cancer (CC) patients and the trend towards more toxic multimodal therapy have led to focus on the quality of life (QOL) of cervical cancer survivors (CCSs). The aim of this critical review was to summarize and discuss the research findings of QOL in CCSs based on self report measures in terms of physical, psychosocial, and sexual well-being. METHODS: Electronic databases were used to identify studies published between 1966 and August 2005. A quality assessment using methodological and treatment related criteria was performed to distinguish between studies with good and less good methodology. RESULTS: Twenty-three studies were included, whereof eight had a good methodology. Eight studies used at least one questionnaire that had not been validated previously, and only one of the validated questionnaires had been tested in former studies of CCSs. The studies with good methodology focused primarily on sexual and social function after treatment, and less on physical and psychological well-being. The trend is that radiotherapy is more associated with reduced QOL dimensions than surgery or chemotherapy. In earlier stages of CC and following surgery alone, there seem to be minor differences between CCSs and control groups concerning various QOL domains. CONCLUSIONS: Reviewed studies indicate that quality of life in cervical cancer survivors is reduced compared to the general female population following radiotherapy, but less so following surgery and earlier stages of cervical cancer. Shortcomings of both methodology and content of the studies reviewed preclude definite conclusions concerning QOL for the moment. PMID- 16716380 TI - Dystocia following prolonged retention of a dead fetus in an Asian elephant (Elephas maximus). AB - A 32-year-old nulliparous female Asian elephant (Elephas maximus) showed signs of parturition 8 months later than predicted from the breeding records. However, while serosanguineous fluid, necrotic tissue and pieces of amnion were expelled, second-stage labor did not progress. Since the fetus was not found during an endoscopic examination of the vestibule, it was assumed that the elephant had calved unseen and she was returned to the forest to recuperate. Twelve months later, the elephant showed clear signs of second-stage labor accompanied by a bulge in the perineum and passage of keratinized nail through the vulva. A 35 cm episiotomy incision was made in the perineum just below the anus, via which chains were attached to the forelimbs of the fetus. Traction on the forelimbs alone proved insufficient to achieve delivery because the fetal head kept rotating and impacting in the pelvis. However, traction applied via a hook inserted behind the mandibular symphysis allowed the head to be elevated and extended, and the fetus to be delivered. The episiotomy wound was sutured in two layers and although the skin did not heal during primary closure it subsequently healed uneventfully by second intention. Retrospective evaluation of the elephant's serum progestagens profile demonstrated a fall to baseline at the suspected onset of parturition, supporting the supposition that the fetus was retained in the uterus for 12 months after parturition began. It is suggested that serum progestagens concentrations should be monitored regularly in mated elephant cows to verify the establishment of pregnancy and to better estimate the expected timing, and the onset of calving. PMID- 16716382 TI - Canine brucellosis: outbreaks and compliance. AB - Canine infertility has many causes that must be considered during evaluation of abnormal reproductive function. An important infectious agent is Brucella canis. Classically deemed a major reason of abortion, this organism also produces infertility in stud dogs and poses a potential health hazard to dogs and humans. The State of Georgia has, out of necessity, instigated regulations to manage outbreaks and seek compliance by educating the pet owner population about this disease. A review of its etiology, methods of transmission, pathophysiology, clinical signs, diagnosis, serology and culture, pathology, treatment options, and regulated prevention featured by Georgia, are presented. PMID- 16716381 TI - Surgical methods of contraception and sterilization. AB - Many techniques for surgically sterilizing dogs and cats have been described; each technique offers advantages and disadvantages to both the patient and surgeon. Techniques that have been described include traditional midline ovariohysterectomy, lateral flank ovariohysterectomy, castration, early age gonadectomy, ovariectomy, laparoscopic ovariohysterectomy and ovariectomy, and vasectomy. Regardless of the technique selected, strict adherence to sound surgical technique and asepsis is mandatory for good surgical outcome with minimal complications. This review will discuss surgical principles, complications, outcomes, as well as relevant current literature associated with each of these techniques of surgical sterilization. PMID- 16716383 TI - Documented and anecdotal effects of certain pharmaceutical agents used to enhance semen quality in the dog. AB - Prostaglandin F(2alpha), gonadotropin releasing hormone, cabergoline and various nutriceuticals have all been recommended by reproductive practitioners to improve sperm motility and morphology and to increase sperm numbers in the ejaculate of the dog. Increasing sperm quantity and quality in the canine ejaculate would benefit all assisted reproductive techniques used in this species. The purpose of this manuscript is to review the documented and anecdotal effects of certain pharmaceuticals used to enhance semen quality in the dog. PMID- 16716384 TI - A special issue in honour of Professor V.M. Shille. PMID- 16716385 TI - Ultrasonography for pregnancy diagnosis and evaluation in queens. AB - In the present paper, we describe the clinical utility of ultrasonography for diagnosing and evaluating pregnancy in domestic cats. Ultrasonography is a non invasive technique that permits an accurate diagnosis of pregnancy and allows serial evaluation of the developing embryo/fetus and the extrafetal structures. The first ultrasonographic indication of pregnancy is a gestational chamber seen on day 10 after mating as a small circular anechoic structure. From day 30, it is possible to recognize different fetal organs, and between 38 and 43 days, the gender of the fetus can be determined. Measurements obtained during the second half of gestation can be used to determine fetal age and calculations can then be made that may more accurately predict the time of parturition. Further studies are needed in the queen to determine the applicability of the echo-Doppler technique used routinely in human obstetric medicine. This type of ultrasonography could potentially provide useful information about fetal health and the maturity of the placenta. PMID- 16716386 TI - Reproductive losses caused by bovine viral diarrhea virus and leptospirosis. AB - Bovine viral diarrhea virus and Leptospira spp. are two of the common pathogenic organisms responsible for reproductive losses in cattle worldwide. Both can be come endemic in herds resulting in chronic low-grade reproductive losses or they can be introduced into relatively naive herds, resulting in substantial reproductive losses over a short period of time. Both organisms are a differential diagnoses for common reproductive losses that veterinarians investigate, including low conception rates and abortions. PMID- 16716387 TI - Neuroanatomically separable effects of imageability and grammatical class during single-word comprehension. AB - The present study characterizes the neural correlates of noun and verb imageability and addresses the question of whether components of the neural network supporting word recognition can be separately modified by variations in grammatical class and imageability. We examined the effect of imageability on BOLD signal during single-word comprehension of nouns and verbs. Subjects made semantic similarity judgments while undergoing functional magnetic resonance imaging (fMRI). Nouns and verbs were matched on imageability, and imageability varied continuously within a grammatical category. We observed three anatomically separable effects: a main effect of grammatical class, a main effect of imageability, and an imageability by grammatical class cross-over interaction. The left superior parietal lobule and a region in the left fusiform responded similarly to increases in noun and verb imageability; the left superior temporal gyrus showed greater activity for verbs than nouns after imageability was matched across grammatical class; and, in both the left middle temporal gyrus and the left inferior frontal lobe, a decrease in noun but not verb imageability resulted in higher BOLD signal. The presence of reliable and anatomically separable main effects of both imageability and grammatical class renders unlikely the hypothesis that previously reported dissociations between nouns and verbs can be dismissed as imageability effects. However, some regions previously thought to respond to grammatical class or imageability instead respond to the interaction of these variables. PMID- 16716388 TI - Left posterior auditory-related cortices participate both in speech perception and speech production: Neural overlap revealed by fMRI. AB - Recent neuroimaging studies and neuropsychological data suggest that there are regions in posterior auditory cortex that participate both in speech perception and speech production. An outstanding question is whether the same neural regions support both perception and production or whether there exist discrete cortical fields subserving these functions. Previous neurophysiological studies suggest that there is indeed regional overlap between these systems, but those studies used a rehearsal task to assess production. The present study addressed this question in an event-related fMRI experiment in which subjects listened to speech and in separate trials, performed a covert object naming task. Single subject analysis revealed regions of coactivation for speech perception and production in the left posterior superior temporal sulcus (pSTS), left area Spt (a region in the Sylvian fissure at the parietal-temporal boundary), and left inferior frontal gyrus. These results are consistent with lesion data and previous physiological data indicating that posterior auditory cortex plays a role in both reception and expression of speech. We discuss these findings within the context of a neuroanatomical framework that proposes these neural sites are a part of an auditory-motor integration system. PMID- 16716389 TI - Sex differences in cerebral laterality of language and visuospatial processing. AB - Sex differences on language and visuospatial tasks are of great interest, with differences in hemispheric laterality hypothesized to exist between males and females. Some functional imaging studies examining sex differences have shown that males are more left lateralized on language tasks and females are more right lateralized on visuospatial tasks; however, findings are inconsistent. Here we used functional magnetic resonance imaging to study thirty participants, matched on task performance, during phonological and visuospatial tasks. For each task, region-of-interest analyses were used to test differences in cerebral laterality. Results indicate that lateralization differences exist, with males more left lateralized during the phonological task and showing greater bilateral activity during the visuospatial task, whereas females showed greater bilateral activity during the phonological task and were more right lateralized during the visuospatial task. Our data provide clear evidence for differences in laterality between males and females when processing language versus visuospatial information. PMID- 16716390 TI - Modeling the brain-pituitary-gonad axis in salmon. AB - To better understand the complexity of the brain-pituitary-gonad axis (BPG) in fish, we developed a biologically based pharmacodynamic model capable of accurately predicting the normal functioning of the BPG axis in salmon. This first-generation model consisted of a set of 13 equations whose formulation was guided by published values for plasma concentrations of pituitary- (FSH, LH) and ovary- (estradiol, 17alpha,20beta-dihydroxy-4-pregnene-3-one) derived hormones measured in Coho salmon over an annual spawning period. In addition, the model incorporated pertinent features of previously published mammalian models and indirect response pharmacodynamic models. Model-based equations include a description of gonadotropin releasing hormone (GnRH) synthesis and release from the hypothalamus, which is controlled by environmental variables such as photoperiod and water temperature. GnRH stimulated the biosynthesis of mRNA for FSH and LH, which were also influenced by estradiol concentration in plasma. The level of estradiol in the plasma was regulated by the oocytes, which moved along a maturation progression. Estradiol was synthesized at a basal rate and as oocytes matured, stimulation of its biosynthesis occurred. The BPG model can be integrated with toxico-genomic, -proteomic data, allowing linkage between molecular based biomarkers and reproduction in fish. PMID- 16716391 TI - Cloning and expression pattern of peroxisome proliferator-activated receptor alpha in the thicklip grey mullet Chelon labrosus. AB - Aquatic organisms living in coastal and estuarine areas are exposed to diverse contaminants which can cause peroxisome proliferation. Peroxisome proliferators are agonists of peroxisome proliferator-activated receptors (PPARs), members of the nuclear receptor superfamily. We have recently demonstrated expression of the three PPAR isoforms in liver of mullet Chelon labrosus and other fish species by immunohistochemistry. The goal of the present study was first to clone PPARalpha and second to investigate its expression pattern in various tissues of mullet. PCR-based screening of mullet cDNA with PPARalpha specific degenerate primers resulted in amplification, subcloning and sequencing of a 1090 bp cDNA fragment (AY618315) that encodes mullet PPARalpha and exhibits highest amino acid identity to fish Sparus aurata PPARalpha (90%). Semi-quantitative RT-PCR was used to characterize the expression of PPARalpha in brain, muscle, liver, spleen, gill, heart and female gonad of juvenile and adult male and female mullet. For this, mullet 18S-rRNA (AY825252), beta-actin (AY836368) and elongation factor alpha (AY836369) were cloned and used as internal reference for RT-PCR. Expression of PPARalpha was detected in all tissues, was highest in liver and lowest in adult male and female muscle. PMID- 16716392 TI - Cloning of Ki-ras and Ha-ras cDNAs from the hermaphroditic fish Rivulus marmoratus (Cyprinodontiformes, Rivulidae) and its expression after exposure to 4 nonylphenol. AB - Previous studies on ras proto-oncogene genes in fish have been focused on chemical-associated carcinogenesis, and the expression of fish ras genes was not well-characterized. We investigated Ki- and Ha-ras genes from the hermaphroditic fish Rivulus marmoratus to understand better their expression patterns in specific tissues, as well as their responses to endocrine-disrupting chemicals such as 4-nonylphenol (4-NP). By investigating expression patterns, we found that the R. marmoratus Ki-ras (Rm Ki-ras) gene showed an alternative splicing event between exons 4A and 4B according to tissue types, which is different from the expression pattern of mammalian Ki-ras genes. In the Rm Ki-ras gene, there were two different expressed types, with exons 1-2-3-4A-4B (long form) and with exons 1-2-3-4B (short form). In the Rm Ki-ras gene, the long form was expressed strongly in the gonad and intestine, and the short form was expressed ubiquitously, except for a low level of expression in the liver. Following 4-NP exposure (300 microg/L), the Rm Ki-ras long form in the liver was significantly expressed, while it was expressed moderately in the ovaries. However, the Rm Ha ras gene was significantly over-expressed in the brain, while its expression in the gonad was down-regulated. In relation to these modulations after 4-NP exposure, we searched the Rm Ha- and Ki-ras promoter regions and found several ERE-half sites, that may be involved in the modulation of ras gene expression following 4-NP exposure. These genes could be applicable as new biomarker genes for assessing exposure to endocrine-disrupting chemicals (EDCs). Further, this implies the disturbance of ras-dependent signal transduction following EDC exposure. PMID- 16716393 TI - Development of new-tools to investigate toxicological hazard due to endocrine disruptor organochlorines and emerging contaminants in Mediterranean cetaceans. AB - The possibility that certain Mediterranean cetaceans are subject to toxicological hazard due to organochlorines and emerging contaminants, such as polybrominated diphenyl ethers (PBDEs) with endocrine disrupting capacity, was investigated using non-lethal methods. The need for new biomarkers for EDCs and for a "cell model" to explore the different susceptibilities to several classes of ECDs, including emerging contaminants, led us to culture fibroblasts of different cetacean species ("dolphins in test tubes"). We then explored interspecies and gender susceptibility to OC-EDCs and PBDEs using qualitative and semi quantitative evaluation of target proteins, such as CYP1A and CYP2B in cultured cetacean fibroblasts (Stenella coeruleoalba, Tursiops truncatus and Balaenoptera physalus), by western blot and immunofluorescence techniques. PMID- 16716394 TI - Heme oxygenase induction and biliverdin excretion: implications for the bile fluorescence biomarker. AB - The measurement of bile fluorescence has become a popular biomarker to demonstrate the exposure of fish to polycyclic aromatic hydrocarbons. Conflicting data have been published on how to normalize bile fluorescence. To investigate if normalization to biliverdin is a suitable method, experiments were performed to study the mechanisms related to biliverdin excretion in fish. Channel catfish (Ictalurus punctatus) were dosed with mixtures of benzo[a]pyrene and cadmium, chlorinated phenols or borneol. The results showed that under increasing toxicant stress, more biliverdin was excreted per amount of protein. To investigate if the increased biliverdin excretion was related to increased heme degradation, enzymatic activity of heme oxygenase (HO) was measured in liver homogenates. The fish dosed with chemical mixtures had significantly higher HO activity than the control fish, and a significant correlation was observed between HO activity and biliverdin concentration in the bile. It is concluded that chemical mixtures of environmental pollutants can induce HO activity and that this chemical stress leads to increased biliverdin excretion. The elucidation of this mechanistic pathway warrants that bile fluorescence is better expressed per amount of bile protein than per biliverdin absorption. PMID- 16716395 TI - Nanoparticulate drug carriers based on hybrid poly(D,L-lactide-co-glycolide) dendron structures. AB - We describe a general method for incorporating target moieties in a well-defined arrangement into the surface of biocompatible polyester poly(D,L-lactic-co glycolic acid) (PLGA) materials using dendrons. In this way it is possible to obtain nanoparticles (NPs) with a high degree of surface coverage. This new strategy was successfully applied to the preparation of peptide- and beta-D glucose-covered NPs. The first application is based on the discovery of NPs made of conjugates between PLGA and short peptidic sequences able to cross the blood brain barrier (BBB) after systemic administration. In this paper, we used a branched structure (dendron) in order to prepare a derivative of PLGA able to form, by simple nanoprecipitation, NPs with a higher degree of surface coverage than previously reported by us, characteristic that could influence the uptake by the liver and spleen. The NPs thus obtained retain the ability to cross the BBB and possess a core-shell structure, as evidenced from zeta-potential, X-ray photoelectron (ESCA) spectroscopy and elemental analyses. These results are comparable with the NPs obtained by the derivatization of preformed NPs. The same strategy, namely the use of a branched spacer (a dendron or a G1 dendrimer) inserted between one end of the PLGA chain and a derivatizing molecule, was also successfully applied to obtain beta-D-glucose-covered NPs; in this case, the surface analysis of the NPs was performed by using high resolution magic angle spinning (HRMAS) NMR spectroscopy and zeta-potential measurements. PMID- 16716396 TI - Monitoring cell adhesion on tantalum and oxidised polystyrene using a quartz crystal microbalance with dissipation. AB - The quartz crystal microbalance with dissipation (QCM-D) (Q-Sense AB, Sweden) has been established as a useful tool for evaluating interactions between various biological and non-biological systems, and there has been increasing interest in using the QCM-D technique for cell monitoring applications. This study investigated the potential of the QCM-D to characterise the initial adhesion and spreading of cells in contact with protein precoated biocompatible surfaces. The QCM-D technique is attractive for monitoring cell adhesion and spreading as it allows in situ real-time measurements. The adhesion of NIH3T3 (EGFP) fibroblasts to tantalum (Ta) and oxidised polystyrene (PS(ox)) surfaces precoated with serum proteins was examined using the QCM-D for a period of either 2 or 4 h. Time-lapse photography was performed at 30 min intervals to visually examine cell adhesion and spreading in order to relate cell morphology to the QCM-D response. Following adsorption of albumin, fibronectin or newborn calf serum onto the surfaces, QCM-D measurements showed that cells adhered and spread on the fibronectin and serum coated surfaces, while few cells adhered to the albumin coated surfaces. Cells adhered to albumin coated surfaces had a rounded morphology. The responses to fibronectin and serum precoated surfaces were quite different for each of the underlying substrates indicating that the process of cell adhesion and spreading elicits different responses depending on both the protein coating composition and the influence of the underlying substrate. The different response may be due to extracellular matrix remodelling as well as cytoskeletal changes. Frequency (f) and dissipation (D) changes associated with cell adhesion were less than would be expected from the Sauerbrey relation due to the viscoelastic properties of the cells. PMID- 16716397 TI - Fludarabine-induced autoimmune hemolytic anemia in a CLL patient: rituximab as the treatment of choice? PMID- 16716398 TI - Lactational exposure to malathion inhibits brain acetylcholinesterase in mice. AB - The organophosphorus (OP) pesticide malathion is a highly neurotoxic compound. Although some studies have reported neurotoxicity signs after the in utero exposure to OP pesticides, there is no evidence of the exclusive contribution of the lactational exposure to malathion as a possible cause of neurotoxicity in the offspring. In this study, we investigated the exclusive contribution of malathion exposure through maternal milk on the activity of acetylcholinesterase (AChE), as well as on biochemical parameters related to the oxidative stress (glutathione levels, lipid peroxidation and glutathione reductase and glutathione peroxidase activities) in the brain of suckling mice. The same parameters were also evaluated in the brains of the respective mothers, which where directly exposed to malathion during the lactational period (daily s.c. injections; doses of 20, 60 and 200mg/kg of body weight). Our results showed that the lactational exposure to malathion caused a high inhibitory effect of the brain AChE activity in the offspring, even when dams were exposed to the lowest malathion dose (20mg/kg). Brain AChE activity was also inhibited in mothers; however, only at the highest malathion dose (200mg/kg). No changes were observed in the biochemical parameters related to the oxidative stress for both dams and pups brains. The present study shows, for the first time, that the exposure of neonatal mice to malathion via lactation inhibits the activity of brain AChE in the offspring. These data, summed to the fact that OP pesticides are excreted in human milk, makes relevant the lactational exposure to these xenobiotics in terms of human health concerns. PMID- 16716399 TI - Development and validation of a test battery to assess subtle neurodevelopmental differences in children. AB - There is increasing concern over the impact of low-dose exposures to environmental chemicals on children's neurobehavioral function. To determine subtle alterations in children's function, it is necessary to move beyond global measures such as IQ and employ tests that can detect small, subtle neurodevelopmental effects across a broad array of behavioral domains. We investigated the sensitivity and specificity of a battery of 63 neurodevelopmental tests or tasks designed to detect outcomes representing the type of subtle neurodevelopmental deficits caused by exposure to neurotoxicants in school-aged children. We studied Neonatal Intensive Care Unit (NICU) graduates, a population known to be at risk for both major and mild anomalies in perception, motor functioning, learning, memory and cognition. This population served as a surrogate to evaluate the capacity of these tests and tasks to predict such deficits. The subjects' histories of previous exposures to any environmental neurotoxicants was not ascertainable, but exposures to elevated levels was not suspected. Over one-third of the 63 measures proved capable of detecting pre-diagnosed lower IQ, the presence of a learning disability (LD) or a neonatal risk profile with at least 70% sensitivity and specificity. Some tests were differentially sensitive and specific, depending upon the presence or absence of one or more of several covariates such as gender, age, hearing status, or familiarity with computers. Tests were also eliminated from the battery if they were affected by too many covariates. We propose calling the final battery of tests that are specific and sensitive to subtle neurodevelopmental changes the Rochester test battery (RTB). Further studies are needed to confirm the capability of the RTB to detect subtle changes associated with neurotoxic exposures. PMID- 16716400 TI - The association between neurobehavioral performance and lung function. AB - This study investigated the relationship between neurobehavioral performance and pulmonary function in young and middle adults. Data were collected from 178 subjects (88 male, 90 female) aged 20-50 years. Neurobehavioral performance was assessed by the Korean version of The Swedish Performance Evaluation System (SPES K), on five items: simple reaction time, symbol digit, digit span, and finger tapping speed for dominant and non-dominant hands. Pulmonary function, as indicated by the 1-s forced expiratory volume (FEV1), was measured according to American Thoracic Society criteria and the potential association was analyzed by linear regression. For simple reaction time, symbol digit, and digit span, the performance of subjects with FEV1 levels exceeding 3.29 l was 2-4% greater than those with the lowest levels of less than 2.60 l, after controlling for the effects of age, sex, education, smoking, and height. Finger tapping speed, however, did not show any significant association. These findings suggest that pulmonary function may influence neurobehavioral performance, especially cognitive abilities, throughout adulthood. PMID- 16716401 TI - Iron-sulfur clusters: why iron? AB - This communication addresses a simple question by means of density functional calculations: Why is iron used as the metal in iron-sulfur clusters? While there may be several answers to this question, it is shown here that one feature - the well-defined inner-sphere reorganization energy of self-exchange electron transfer - is very much favored in iron-sulfur clusters as opposed to metal substituted analogues of Mn, Co, Ni, and Cu. Furthermore, the conclusion holds for both 1Fe and 2Fe type iron-sulfur clusters. The results show that only iron provides a small inner-sphere reorganization energy of 21 kJ/mol in 1Fe (rubredoxin) and 46 kJ/mol in 2Fe (ferredoxin) models, whereas other metal ions exhibit values in the range 57-135 kJ/mol (1Fe) and 94-140 kJ/mol (2Fe). This simple result provides an important, although partial, explanation why iron alone is used in this type of clusters. The results can be explained by simple orbital rules of electron transfer, which state that the occupation of anti-bonding orbitals should not change during the redox reactions. This rule immediately suggests good and poor electron carriers. PMID- 16716402 TI - Clinical significance and distribution of putative virulence markers of 116 consecutive clinical Aeromonas isolates in southern Taiwan. AB - OBJECTIVES: The objectives of this study were to elucidate the clinical manifestations of Aeromonas infections and the association of putative virulence genes with clinical invasiveness. METHODS: 116 consecutive clinical Aeromonas isolates collected from July 1999 to June 2001 in a medical center in southern Taiwan were included. All isolates were identified by biochemical phenotyping and their genomic sequences encoding eight putative virulence factors, including cytolytic enterotoxin (AHCYTOEN), aerolysin (aerA), hemolysin (hlyA), heat-labile enterotoxin (alt), heat-stable enterotoxin (ast), and components of type III secretion system (ascV, aexT and ascF-ascG) were analyzed using polymerase chain reaction and colony blot hybridization. The association of clinical diseases of the patients with the putative virulence genes in the isolates was analyzed. RESULTS: Sixty-two percent of Aeromonas isolates caused clinically evident infections, of which the major clinical manifestations were primary bacteremia (40%), followed by soft tissue infections (27%), and hepatobiliary tract infections (15%). Liver cirrhosis (36%), malignancy (25%), and hepatobiliary diseases (13%) were the major underlying diseases in patients with Aeromonas bacteremia. The majority (64%) of patients with Aeromonas hepatobiliary infections had underlying hepatobiliary diseases, whereas 71% of those with soft tissue infections had antecedent water- or soil-related injuries. The crude fatality rate for Aeromonas infections was 26%. Aeromonas hydrophila complex was the most common (52%) of the three major complex groups investigated, followed by Aeromonas sobria complex (24%) and Aeromonas caviae complex (23%). None of the eight putative virulence factors was associated with invasiveness or bacteremia. CONCLUSIONS: Primary bacteremia, soft tissue infections, and hepatobiliary tract infections are the three major clinical manifestations of invasive Aeromonas infections in southern Taiwan. This study found no association between the presence of AHCYTOEN, aerA, hlyA, alt, ast, ascV, aexT or ascF-ascG genes in Aeromonas isolates and the development of extra-intestinal infections or bacteremia. PMID- 16716403 TI - Arguments pro disinfection in diagnostic electron microscopy: a response to Madeley and Biel. PMID- 16716404 TI - Pancreatic abscess due to Aeromonas hydrophila. AB - Aeromonas hydrophila is an uncommon cause of deep-seated infection in man. We present what we believe to be the first reported case of a pancreatic abscess due to A. hydrophila. The outcome of this case attests to the potentially lethal nature of infection that can occur with this organism. PMID- 16716405 TI - Atypical cutaneous manifestation of HSV-2 with Candida albicans co-infection in a patient with HIV-1. AB - Herpes simplex virus type 2 (HSV-2) infection was one of the first opportunistic infections identified among patients with AIDS. In the literature there are many data suggesting that the natural history of HSV-2 infection is altered in HIV-HSV 2 co-infected patients. Furthermore, a relationship between HIV seropositivity and HBV infection because of their analogous way of transmission is also described. We report the case of a 37-year-old patient who suffered from multiple painful ulcerative lesions of the perianal region. Laboratory examination showed positivity for HIV and HBV infections. In HIV-positive patients perianal HSV-2 can have atypical manifestations, especially if co-infection by Candida albicans occurs. PMID- 16716406 TI - Therapeutic potential of treating chronic obstructive pulmonary disease (COPD) by neutralising granulocyte macrophage-colony stimulating factor (GM-CSF). AB - Chronic obstructive pulmonary disease (COPD) is a major incurable global health burden and will become the third largest cause of death in the world by 2020. It is currently believed that an exaggerated inflammatory response to inhaled irritants, in particular cigarette smoke, causes progressive airflow limitation. This inflammation, where macrophages and neutrophils are prominent, leads to oxidative stress, emphysema (loss of lung structure), small airways fibrosis and mucus hypersecretion. However, COPD responds poorly to current anti-inflammatory treatments including potent glucocorticosteroids, which produce little or no benefit. In this review we consider the therapeutic potential of targeting granulocyte macrophage-colony stimulating factor (GM-CSF) for the treatment of COPD. GM-CSF is a major regulator of both macrophage and neutrophil activation and survival in the lung-these cells are intimately linked to COPD. Animal data indicates that neutralisation of GM-CSF ameliorates experimental COPD and predicts therapeutic utility in treating stable COPD and treating exacerbations. As such, GM-CSF represents an attractive therapeutic target for the treatment of COPD. PMID- 16716407 TI - Periodically interrupted amperometry. A way of improving analytical performance of membrane coated electrodes. AB - Amperometry is a powerful voltammetric measuring method. Its application is specially advantageous when used in combination with a separation step or with some other sample treatment method providing selectivity. The selectivity is often achieved by coating the amperometric working electrode surface with a membrane of special character. Size exclusion membrane, immobilized enzyme containing reaction layer, protecting dialysis membrane, perm selective ion exchange film etc can be mentioned here. In conventional amperometry the measuring potential is continuously applied, therefore in case of membrane coated electrodes the electrode process depletes the diffusion layer. In this work the performance of a new periodically interrupted amperometric (PIA) measuring program has been investigated in case of glucose enzyme sensor. The measuring program allowing time for reloading the diffusion layer provided higher current and therefore improved sensitivity and lower limit of detection. PMID- 16716408 TI - Topographic differences in mean computational sleep depth between healthy controls and obstructive sleep apnoea patients. AB - In this work, topographic differences in computational sleep depth between healthy controls and obstructive sleep apnoea syndrome (OSAS) patients have been examined. Sleep depth estimation was based on continuous monitoring of the mean frequency of the EEG. During the experiments, all-night sleep EEG recordings of carefully age and gender matched sets of 16 healthy controls and 16 OSAS patients were compared on six electrode locations (Fp1-M2, Fp2-M1, C3-M2, C4-M1, O1-M2, and O2-M1). To optimise the diagnostic ability of the method, we examined the influence of 45 sets of adjustable analysis parameters on the ability of the method to show differences in computational sleep depth between the diagnostic groups. The results show clearly that although the visual scores for a set of epochs are the same for both clinical groups, computational sleep depth measure still shows deeper local sleep for healthy controls, both during NREM and REM sleep. Although the best achievable performance in different sleep stages is reached in different EEG derivations and with different parameter values, computation of sleep depth with 1-s output resolution in non-overlapping segments of 2s (400 samples) with maximum analysis band frequency of 20.5 Hz and 51-point moving median smoothing on Fp2-M1 or O1-M2 leads to near-optimal performance in deep sleep or wakefulness/light sleep, respectively. PMID- 16716409 TI - The antigen recognized by MOMA-I is sialoadhesin. AB - The monoclonal antibody MOMA-1 is a widely-used marker for marginal metallophilic macrophages in spleen and some other subsets of macrophages. The antigen recognized by MOMA-1 has yet to be characterized, but its expression pattern is similar to that of sialoadhesin (Sn, CD169, Siglec-1), a member of the sialic acid binding Ig-like lectin (Siglec) family. Using flow cytometry of Sn transfected cells and staining of lymphoid tissue sections from Sn-deficient mice, we demonstrate here that the antigen recognized by MOMA-1 is Sn. PMID- 16716410 TI - Increased levels of the CD40:CD40 ligand dyad in the cerebrospinal fluid of rats with vitamin B12(cobalamin)-deficient central neuropathy. AB - The levels of the soluble (s) CD40:sCD40 ligand (L) dyad, which belongs to the tumor necrosis factor (TNF)-alpha:TNF-alpha-receptor superfamily, are significantly increased in the cerebrospinal fluid (CSF), but not the serum of cobalamin (Cbl)-deficient (Cbl-D) rats. They were normalized or significantly reduced after treatment with Cbl, transforming growth factor-beta1 or S-adenosyl L-methionine, and the normal myelin ultrastructure of the spinal cord was concomitantly restored. The concomitance of the two beneficial effects of these treatments strongly suggests that the increases in CSF sCD40:sCD40L levels may participate in the pathogenesis of purely myelinolytic Cbl-D central neuropathy in the rat. In keeping with this, an anti-CD40 treatment prevented myelin lesions. PMID- 16716411 TI - Design, development and evaluation of a competitive RT-PCR for quantitation of GBV-C RNA. AB - Although GB virus C (GBV-C) hepatocyte pathogenicity is still controversial, it appears that at least some strains of this virus are lymphotropic. During the past few years, several reports have documented an apparently beneficial role played by GBV-C in the course of HIV-1 infection. At present, a commercial kit for GBV-C RNA quantitation is not available. In this study, a competitive RT-PCR method for GBV-C in serum samples is described. The sensitivity of the assay proved to be 10(4) and 10(3) genomic equivalents for positive and negative sense RNAs, respectively. This method will discriminate specifically between positive and negative strand RNAs with a discrimination index of at least five log10. Out of 60 samples from different hematological disorders (n = 49), HIV-1 positive patients (n = 7), and blood donors (n = 4), 10 proved to be GBV-C RNA positive. Viral load ranged from 1.1 x 10(7) to 2.34 x 10(8) genomic equivalents/ml. Such values correlated linearly (r = 0.986) with those obtained by a 10-fold serial dilution method. In studies exploring the GBV-C pathogenicity, the measurement of viral load may contribute to understand the possible mechanisms involved. PMID- 16716412 TI - Importance of virus-medium interactions on the biological activity of wild-type Heliothine nucleopolyhedroviruses propagated via suspension insect cell cultures. AB - In vitro serial passaging of nucleopolyhedroviruses often results in virus instability, leading to reduction of both yield and biological activity of polyhedra (virus occlusion bodies). In this study, uncloned Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (HaSNPV) and cloned Helicoverpa zea single-nucleocapsid nucleopolyhedrovirus (HzSNPV) were each serially passaged five times in both low cost (VPM) and commercial (Excell 401) media using H. zea cell cultures. When the experimental data was analysed as a 2(3) full factorial design (testing two levels of virus, medium and passage number), the passage number was shown to have the most significant effect on polyhedra yield, while the virus-medium interaction had the most important effect on polyhedra biological activity. This interaction was most pronounced for HzSNPV, which experienced a dramatic decline in biological activity when switched from its original Excell 401 medium to the in-house VPM medium. This suggests that genetically homogeneous viruses are less able to adapt to perturbations in the nutrient environment, hence medium changes should be avoided. Therefore, it is important to standardise the scale-up process of nucleopolyhedrovirus biopesticides at the earliest stage of development, especially when it requires both a low cost medium and a plaque purified virus. PMID- 16716413 TI - The neural substrates of cognition. PMID- 16716414 TI - Antiviral therapy of chronic hepatitis B. AB - Treatment of chronic hepatitis B remains a clinical challenge. Long-term viral suppression is a major goal of antiviral therapy to improve the clinical outcome of the patients. Antiviral treatment of chronic hepatitis B relies currently on immune modulators such as interferon alpha and its pegylated form, and viral polymerase inhibitors. Because of the slow kinetics of viral clearance and the spontaneous viral genome variability, viral mutants resistant to nucleoside analogs may be selected. However, the development of new antiviral agents is rapidly improving the offing of therapy of chronic hepatitis B. These new therapeutic advances are reviewed in this manuscript. PMID- 16716415 TI - Pharmacology of current and promising nucleosides for the treatment of human immunodeficiency viruses. AB - Nucleoside antiretroviral agents are chiral small molecules that have distinct advantages compared to other classes including long intracellular half-lives, low protein binding, sustained antiviral response when a dose is missed, and ease of chemical manufacture. They mimic natural nucleosides and target a unique but complex viral polymerase that is essential for viral replication. They remain the cornerstone of highly active antiretroviral therapy (HAART) and are usually combined with non-nucleoside reverse [corrected] transcriptase and protease inhibitors to provide powerful antiviral responses to prevent or delay the emergence of drug-resistant human immunodeficiency virus (HIV). The pharmacological and virological properties of a selected group of nucleoside analogs are described. Some of the newer nucleoside analogs have a high genetic barrier to resistance development. The lessons learned are that each nucleoside analog should be treated as a unique molecule since any structural modification, including a change in the enantiomeric form, can affect metabolism, pharmacokinetics, efficacy, toxicity and resistance profile. PMID- 16716416 TI - Chronic exposure of rats to noise: relationship between long-term memory deficits and slow wave sleep disturbances. AB - Noise is now recognized as a serious health problem in our modern societies. Although its deleterious and direct effects on cognitive tasks (long-term memory, mental arithmetic activity, visual tasks, etc.) are clearly admitted, no studies have determined a delayed indirect effect of noise on cognitive processes. Furthermore, the link between sleep disturbances related to environmental noise (EN) exposure and these indirect deteriorations of human performances has never been demonstrated. This could be due to inappropriate evaluation of sleep as well as to uncontrolled and confounding factors such as sex, age, and also inter individual vulnerability. Based on a recently validated animal model [Rabat A, Bouyer JJ, Aran JM, Le Moal M, Mayo W. Chronic exposure to an environmental noise permanently disturbs sleep in rats: inter-individual vulnerability. Brain Res 2005;1059:72-82], aims of the present study were (i) to determine long-term memory (LTM) deficits following a chronic exposure to EN and (ii) to link these behavioral problems to sleep disturbances related to EN. For this purpose in a first experiment, LTM performances were evaluated before and following 9 days of EN. Results show LTM deficits following a chronic exposure to EN with inter individual vulnerability. Vulnerability profile was related to the psychobiological profile of rats. Results of the second experiment show LTM deficits correlated to both debt of slow wave sleep (SWS) and to daily decrease of SWS bout duration. Our results demonstrate that chronic exposure to noise indirectly disturbs LTM possibly through SWS disturbances and suggest a possible role of the stress hormonal axis in these biological effects of noise. PMID- 16716417 TI - Olanzapine differentially affects 5-HT2Aand2C receptor mRNA expression in the rat brain. AB - This study examined regional changes in rat brain mRNA levels encoding 5-HT(2A) and 5-HT(2C) receptors following chronic olanzapine treatment. The immediate effect (2h after the last treatment) was a down-regulation of 5-HT(2A) receptor mRNA expression, predominantly in the hypothalamus, limbic system and striatum, while a rebound effect was observed 48 h later. 5-HT(2C) receptor mRNA expressions were decreased in the substantia nigra. Correlations between 5-HT(2A) receptor mRNA expression and total food intake, weight gain and energy efficiency were observed. PMID- 16716418 TI - Transient transfection of Plasmodium vivax blood stage parasites. PMID- 16716419 TI - EhPAK2, a novel p21-activated kinase, is required for collagen invasion and capping in Entamoeba histolytica. AB - p21-activated kinases (PAKs) are a highly conserved family of enzymes that are activated by Rho GTPases. All PAKs contain an N-terminal Cdc42/Rac interacting binding (CRIB) domain, which confers binding to these GTPases, and a C-terminal kinase domain. In addition, some PAKs such as Cla4p, Skm1p and Pak2p contain an N terminal pleckstrin homology (PH) domain and form a distinct group of PAK proteins involved in cell morphology, cell-cycle and gene transcription. Here, we describe a novel p21-activated kinase, denominated EhPAK2, on the parasitic protozoan Entamoeba histolytica. This is the first reported Entamoeba PAK member that contains a N-terminal PH domain and a highly conserved CRIB domain. EhPAK2 CRIB domain shares 29% of amino acid identity and 53% of amino acid homology with these of DdPAKC from Dictyostelium discoideum and Cla4p from Saccharomyces cerevisiae and binds in vitro and in vivo to EhRacA GTPase. This domain also possesses the conserved residues His123, Phe134 and Trp141, which are important for the interaction with the effector loop and strand beta2 of the GTPase; and the residues Met121 and Phe145, which are specific for the interaction of EhPAK2 with EhRacA. Functional studies of EhPAK2 showed that its C-terminal kinase domain had activity toward myelin basic protein. Cellular studies showed that Entamoeba trophozoites transfected with the vector pExEhNeo/kinase-myc, had a 90% decrease in the ability to invade a collagen matrix as well as severe defects in capping, suggesting the involvement of EhPAK2 in these cellular processes. PMID- 16716420 TI - Biomimetic approaches to protein and gene delivery for tissue regeneration. AB - Novel therapeutic strategies that promote wound healing seek to mimic the response of the body to wounding, to regenerate rather than repair injured tissues. Many synthetic or natural biomaterials have been developed for this purpose and are used to deliver wound therapeutics in a controlled manner that prevents unwanted and potentially harmful side-effects. Here, we review the natural and synthetic biomaterials that have been developed for protein and gene delivery to enhance tissue regeneration. Particular emphasis is placed on novel biomimetic materials that respond to environmental stimuli or release their cargo according to cellular demand. Engineering biomaterials to release therapeutic agents in response to physiologic signals mimics the natural healing process and can promote faster tissue regeneration and reduce scarring in severe acute or chronic wounds. PMID- 16716421 TI - Mitochondrial membrane potential and ischemic neuronal death. AB - Mitochondria are intracellular organelles in which high energy phosphate is produced. Ischemia causes depletion of the materials necessary to produce this phosphate and strongly affects the electron transport chain. Apoptosis commences during and after ischemia. As such, it is likely that a significant relationship exists between inactivation of electron transport and apoptosis. Mitochondrial membrane potential (MMP) reflects performance of the electron transport chain and can indicate a pathological disorder of this system. In an experimental setting, oxygen-glucose depletion (OGD) in neuronal cell culture has been employed to simulate an ischemic condition. The relationship between MMP and subsequent neuronal death during and after OGD has been examined. MMP dissipation and concomitant neuronal death have been reported, but recent studies have demonstrated mitochondrial hyperpolarization preceding neuronal death. The direction of MMP polarization depends on the extent of OGD. Long OGD results in depolarization, while shorter OGD induces hyperpolarization. Neurons are still viable during hyperpolarization, but the process may switch on the apoptotic cascade. Meanwhile, dissipation of MMP seems to be a consequence of severe energy deficit, leading to necrosis. MMP may be a marker of subsequent apoptosis, although a causal relationship remains to be determined. PMID- 16716422 TI - Pattern differentiation of excitatory and inhibitory synaptic inputs on distinct neuronal types in the rat caudal nucleus of the tractus solitarius. AB - Region- and size-specific neuronal organizations of the caudal nucleus of the tractus solitarius (cNTS) were investigated, followed by analyses of excitatory and inhibitory synaptic input patterns onto specific cell types by patch clamp recordings and immunoelectron microscopy. Cell size distribution and numerical density of cNTS neurons were examined in subregions at levels of the area postrema. In the subpostremal and dorsomedial subnuclei, characterized by the presence of dense glutamatergic and sparse GABAergic somata, small calbindin neurons constituted 42% of the total cells. The medial subnucleus contained large numbers of glutamatergic, GABAergic, and catecholaminergic somata and large tyrosine hydroxylase-containing cells constituted 13% in this region. In total, small neurons (<150 microm2) represented about 80% of the cell population in the cNTS. Predominant excitatory postsynaptic currents were observed in the adult small neurons, while inhibitory postsynaptic currents were more evident in larger neurons, irrespective of subnuclear location. This distinct differentiation of postsynaptic current patterns was not evident in neonates. GABAergic synapses were more frequently associated with dendrites of large catecholaminergic cells (73%) than with those of small calbindin-containing cells (10%) in adults. These results indicate that differential synaptic input patterns were developmentally established in distinct small and large neurons. PMID- 16716423 TI - Listeria innocua and Lactobacillus delbrueckii subsp. bulgaricus employ different strategies to cope with acid stress. AB - Responses of Listeria innocua and Lactobacillus delbrueckii subsp. bulgaricus to a rapid change in extracellular pH (pHex) from pHex 6 to a range of concentrations down to pHex 3.0 were examined, using HCl and lactic acid (LA) as acidulants. A new fluorescent probe 5-(and-6)-carboxy-2', 7'-dichlorofluorescein diacetate succinimidyl ester (CDCFDA-SE) was employed that enabled reliable measurements of intracellular pH (pHi) to a minimum pHi of 4.0. Changes in pHi and H+ fluxes from immobilised bacteria were measured using fluorescence ratio imaging microscopy (FRIM) and a non-invasive ion flux measuring technique (MIFE), respectively. L. innocua maintained a relatively constant pHi of 5.5-6.1 at pHex 4 and 5 via H+ extrusion. In contrast, L. delbrueckii subsp. bulgaricus progressively lowered pHi towards pHex over the entire pHex range examined. The type of acidulant used influenced pH regulation with both pHi and H+ -fluxes being more severely affected by LA compared to HCl. Overall, our data demonstrated different adaptive strategies in these two bacteria. While L. innocua expels protons to maintain a constant pHi, L. delbrueckii subsp. bulgaricus allows proton entry after acidic treatment so that pHi follows pHex. PMID- 16716424 TI - Occurrence of ochratoxin A in wine and ochratoxigenic mycoflora in grapes and dried vine fruits in South America. AB - Grape and wine production in South America represents about 6.6% and 10% respectively of the world grape and wine production. The available information on the ochratoxigenic mycoflora and ochratoxin A (OTA) presence in wine grapes, wines, grape juices and dried vine fruits is limited. Surveys have been carried out in Argentina and Brazil which showed that Aspergillus niger aggregate are predominant in the Argentinean varieties while from the Brazilian varieties the species A. niger, Aspergillus ochraceus and Aspergillus carbonarius were isolated. A mycobiota survey from wine grapes in Argentina showed that while Alternaria alternata was predominant, Aspergillus section Nigri species were isolated from 60% of samples. About 41% of black Aspergilli isolates produced OTA with levels ranging from 2 to 24.5 ng mL(-1). In another study, about 83% of A. carbonarius isolates from dried vine fruits produced OTA, with levels ranging from 2 to 5200 ng mL(-1). A survey of grape juices and wines of Brazilian, Argentinean and Chilean origin were found to contain very low levels of OTA. Studies are in progress in Latin America on the ecophysiology of ochratoxigenic fungi and OTA occurrence to reduce the impact of this toxin in the food chain. PMID- 16716425 TI - Molecular identification of the edible ectomycorrhizal fungus Lactarius deliciosus in the symbiotic and extraradical mycelium stages. AB - Specific rDNA ITS amplifications, microsatellite-primed PCR and ITS-SSCP analysis were applied to identify and characterize pre-selected isolates of the edible ectomycorrhizal fungus Lactarius deliciosus in different stages of the life cycle. Sampling was performed from pure cultures, mycorrhizas and soil from experimental plots established with nursery-inoculated pine seedlings. A newly designed reverse primer (LDITS2R) combined with the universal forward ITS1 allowed to perform specific amplifications of L. deliciosus from all the samples. Microsatellite-primed PCR using the (GTG)5 oligonucleotide as a primer showed clear polymorphisms among the different L. deliciosus isolates. The patterns of mycorrhiza samples showed additional bands corresponding to the plant DNA. Single strand conformation polymorphism (SSCP) analysis of the specific rDNA ITS fragment amplified from 18 L. deliciosus isolates showed nine clearly different patterns. Mycorrhiza and soil samples showed coincident patterns with their respective fungal isolates. Specific rDNA ITS amplifications had not been previously used for SSCP analysis of ectomycorrhizas and extraradical mycelium. This relatively simple and inexpensive technique allows tracking L. deliciosus isolates in different stages of the fungus development. Specific ITS-SSCP analysis is promising in studies of the persistence of inoculated L. deliciosus isolates and their competitiveness with native ectomycorrhizal fungi, especially at the extraradical mycelium stage. PMID- 16716426 TI - High level expression of bioactive recombinant human growth hormone in the milk of a cloned transgenic cow. AB - Transgenic farm animals have been proposed as an alternative to current bioreactors for large scale production of biopharmaceuticals. However, the efficiency of both methods in the production of the same protein has not yet been established. Here we report the production of recombinant human growth hormone (hGH) in the milk of a cloned transgenic cow at levels of up to 5 g l(-1). The hormone is identical to that currently produced by expression in E. coli. In addition, the hematological and somatometric parameters of the cloned transgenic cow are within the normal range for the breed and it is fertile and capable of producing normal offspring. These results demonstrate that transgenic cattle can be used as a cost-effective alternative for the production of this hormone. PMID- 16716427 TI - The cytostat: A new way to study cell physiology in a precisely defined environment. AB - In a cytostat, a continuous culture is monitored and controlled by an automated flow cytometer system, based on the determination of the cell concentration and the single cell property distribution of the growing cell population. The growing culture can be maintained at steady state even at such low cell concentrations that the bioreactor medium composition is negligibly changed by the few cells. Therefore, the cell environment is precisely defined by the feed composition since products of cell growth are not present in significant amounts. Effects on cell growth of nutrients, of toxic compounds such as drugs, or of products made by the cells, if added to the feed medium, can be readily isolated. Using the cytostat, it is shown here that ethanol assumes the triggering function for the increase in cell size in Saccharomyces cerevisiae normally only seen at critical growth rates above critical cell densities. This suggests that ethanol assumes a quorum sensing function on cell growth when a critical cell density is reached. PMID- 16716428 TI - Preparative use of isolated CYP102 monooxygenases -- a critical appraisal. AB - Isolated P450 monooxygenases have for long been neglected catalysts in enzyme technology. This is surprising as they display a remarkable substrate specificity catalyzing reactions, which represent a challenge for classic organic chemistry. On the other hand, many P450 monooxygenases are membrane bound, depend on rather complicated electron transfer systems and require expensive cofactors such as NAD(P)H. Their activities are low, and stability leaves much to be desired. The use of bacterial P450 monooxygenases from CYP102 family allows overcoming some of these handicaps. They are soluble and their turnovers are high, presumably because their N-terminal heme monooxygenase and their C-terminal diflavin reductase domain are covalently linked. In recent years, protein engineering approaches have been successfully used to turn CYP102 monooxgenases into powerful biocatalysts. PMID- 16716429 TI - Photoreactive cellulose membrane--A novel matrix for covalent immobilization of biomolecules. AB - We report a simple and mild procedure for the preparation of a photoreactive cellulose membrane capable of forming a covalent bond with a biomolecule in presence of 365 nm UV light. Photoreactive cellulose membrane was prepared by the reaction of fluoro group of 1-fluoro-2-nitro-4-azidobenzene (FNAB) and hydroxyl group of the cellulose in an alkaline medium. X-ray photoelectron spectroscopy (XPS) of the photoreactive cellulose confirmed the incorporation of FNAB moiety. Azido group of the photoreactive membrane on exposure to UV light transforms into highly reactive nitrene which binds with a protein. The efficacy of the activated membrane was checked by immobilizing glucose oxidase (GOD) onto it in presence of light. Immobilized GOD was found to have improved thermal, pH and storage stability. Photoreactive cellulose membrane was successfully used in enzyme linked immunosorbent assay (ELISA) technique. The antibody immobilized onto such support by UV irradiation in 30 min showed similar ELISA value than the antibody immobilized onto a polystyrene ELISA plate in 12h incubation at 4 degrees C by conventional method. PMID- 16716430 TI - Izumoring: a strategy for bioproduction of all hexoses. AB - Izumoring is a new concept for the bioproduction of all hexose sugars - 16 aldohexoses, eight ketohexoses and 10 hexitols -- using enzymatic and microbiological reactions. The biocatalysts necessary for the strategy were (i) d tagatose 3-epimerase [which epimerizes all ketohexoses at C-3 of the sugar], (ii) polyol dehydrogenases [which catalyze oxidation-reduction reactions between ketohexoses and the corresponding hexitols], and (iii) aldose isomerases [which catalyze isomerization reactions between aldohexoses and corresponding aldoketoses]. All ketohexoses, aldohexoses and hexitols may be arranged in a symmetric ring connected by the biochemical reactions, allowing the design for the bioproduction of all 34 hexose sugars. The ring shows there are four entrances to the l-hexose world from the natural d-hexoses. The Izumoring ring shows clearly the relationship and the position of all 34 six carbon sugars. PMID- 16716431 TI - Improving baculovirus transduction of mammalian cells by surface display of a RGD motif. AB - An RGD-containing peptide, comprising 23 amino acids from the foot-and-mouth disease virus (FMDV) VP1 protein was engineered into the envelope of Autographa californica nuclear polyhedrosis virus surface (AcNPV) using two different display strategies. The RGD-motif is a well-described tripeptide, that by binding to cell surface integrins facilitates virus entry into cells. This epitope was displayed, either by directly modifying the native major envelope protein gp64 of AcNPV, or by incorporating a second, modified version of gp64 onto the virus surface. Transduction efficiencies of four mammalian cell lines were compared by detecting the expression of the reporter gene green fluorescent protein (gfp), delivered by the baculovirus genome. Our results showed that insertion of the RGD peptide into the envelope protein gp64 leads to enhanced specific uptake of baculoviral particles in mammalian cells, only when a combination of wild-type and mutant gp64 was present on the viral surface. Whenever the RGD-peptide was directly inserted into the native gp64, the overall amount of gp64 envelope protein was diminished, leading to decreased viral uptake. PMID- 16716432 TI - Advances in cloning, functional analysis and heterologous expression of fungal polyketide synthase genes. AB - Fungal polyketides comprise a diverse group of secondary metabolites that play an important role for drug discovery, as pigments, and as mycotoxins. Their biosynthesis is governed by multidomain enzymes, so-called fungal type I polyketide synthases (PKS). Investigating the molecular basis of polyketide biosynthesis in fungi is of great importance for ecological and pharmacological reasons. In addition, cloning, functional analysis and expression of fungal PKS genes also set the basis for engineering the yet largely untapped biosynthetic potential. PMID- 16716436 TI - Fatigue after stroke. AB - OBJECTIVES: To examine the phenomenon of fatigue after stroke and to review the knowledge about frequency, consequences, associated factors, physiopathology and treatment. MATERIALS AND METHOD: Medline was systematically searched with the following keywords: stroke, fatigue, sleep disorders, exercise, and rehabilitation. All relevant articles found in the references were screened as well. RESULTS AND DISCUSSION: Fatigue is a common complaint after stroke and occurs in 39-72% of stroke survivors. Some studies show a severe functional impact of this symptom as well as a high mortality rate. Available evidence concerning associated factors is limited, but fatigue is clearly multifactorial. Some studies show that limited exercise capacity, increased gait energy cost, sleep-disordered breathing and sleep disorders can be related to physical fatigue. Other studies show a link between fatigue and depression. The existence of primary fatigue is still controversial. Treatment must follow a diagnostic approach. Treadmill training, among other treatments, improves fitness reserve and lowering of the energy cost of hemiparetic gait, which could be useful in relieving fatigue. PMID- 16716433 TI - Fatigue in patients with cardiovascular disease. AB - Fatigue is a frequent complaint during cardiovascular disease and can sometimes constitute the first clinical manifestation of this disease. It is responsible for deterioration of the quality of life and prognosis. Although physical and mental fatigue are often intimately interrelated, these two aspects of fatigue correspond to different pathophysiological mechanisms and different clinical features and the neurobiological links between the two are only just beginning to be studied. Physical fatigue is related to loss of efficacy of the effector muscle, due to multiple causes: mismatch of cardiac output during exercise, muscle and microcirculatory deconditioning, neuroendocrine dysfunction, associated metabolic disorders. Mental fatigue corresponds to predominantly depressive mood disorders with a particular entity, vital exhaustion. The diagnostic approach is designed to eliminate other organic causes of fatigue. Functional tests investigating physical (exercise capacity) and mental dimensions (mood disorders) can be used to analyse their respective roles and to propose personalized management, in which rehabilitation has an essential place due to its global approach. The objective of this reduction of fatigue is threefold: to improve independence, to improve quality of life and to limit morbidity and mortality. PMID- 16716437 TI - Fatigue after spinal cord injury. AB - OBJECTIVES: To identify variables increasing fatigue following spinal cord injury (SCI) and their functional consequences. METHODS: A search of the Medline and Reedoc databases with the keywords SCI, fatigue, intrinsic muscular fatigue, chronic fatigue, aging, training, electrostimulation, quality of life and the same words in French. RESULTS: Two kinds of fatigue are identified following SCI. Intrinsic fatigue in muscles totally or partially paralysed at the level of or below the spinal cord lesion; this peripheral fatigue is due to denervation, total or partial loss of motoneurons, or histological and metabolical changes in muscle; it is well-defined by electrophysiological technology; spasticity and spasms have little influence on its development; it is reversible in part with long term electrostimulation, but at this time, electroneuroprosthetic techniques do not reduce the excessive energetic cost to stand up and walk. Chronic fatigue appears in the long term following SCI; it is linked with aging, physiological, and psychological deconditioning; some data point to chronic fatigue after SCI similar to post-polio syndrome and chronic fatigue syndrome, which may explain the central nature of the fatigue; training programs could be useful in delaying this chronic fatigue and as a consequence, increasing the latent quality of life. CONCLUSION: Muscular intrinsic fatigue after SCI is always of a peripherical nature in muscles partially or totally paralysed. Chronic fatigue during aging greatly decreases quality of life. Both intrinsic and chronic fatigue could be anticipated by electrostimulation technique on the one hand and long term training on the other. PMID- 16716438 TI - Fatigue and traumatic brain injury. AB - Fatigue is frequent and disabling in patients with traumatic brain injury (TBI). Its mechanisms are complex and multifactorial. We performed a literature review of reports of the condition using the following key words: brain injury, depression, neuroendocrine dysfunction, and treatment. Five scales have been used to evaluate fatigue in TBI patients: the Fatigue Severity Scale, the visual analog scale (VAS) for fatigue, the Fatigue Impact Scale, the Barrow Neurological Institute (BNI) Fatigue Scale and the Cause of Fatigue (COF) Questionnaire. The BNI Fatigue Scale and the COF Questionnaire have been designed specifically for brain-injured patients. Fatigue is present in 43-73% of patients and is one of the first symptoms for 7% of them. Fatigue does not seem to be significantly related to injury severity not to time since injury. It can be related to mental effort necessary to overcome attention deficit and slowed processing ("coping hypothesis"). It can also be related to sleeping disorders and depression, although the relation between fatigue and depression are debated. Finally, fatigue can also be related to infraclinical pituitary insufficiency (growth hormone insufficiency, hypocorticism). To date, no published study of treatment of fatigue after TBI exists. PMID- 16716440 TI - [Psychobehavioural assessment for chronic low back pain]. AB - INTRODUCTION: Psychobehavioural assessment may be a complementary approach to assess response to treatment for pain and disability in patients with low back pain. OBJECTIVES: To assess the correlation between psychobehavioural factors and pain and disability in patients with chronic low back pain in France. METHODS: We asked 83 patients with chronic low back pain to complete questionnaires during a primary care consultation by a general practitioner. Pain was measured by a visual analog scale (VAS), disability by the Quebec Back Pain Disability Scale and psychobehavioural factors by the Fear-Avoidance Beliefs Questionnaire (FABQ) and the Coping Strategy Questionnaire (CSQ). RESULTS: Pain was poorly correlated with anxiety (R = 0.36) and scores on the FABQ 1 (R = 0.46) and FABQ 2 (R = 0.30) and not correlated with depression (R = 0.22), duration of pain evolution (R = 0,10) and body mass index (R = 0.12). The duration of stopping work was poorly correlated with disability (R = 0.35) and FABQ 1 score (R = 0.43) and not correlated with pain (R = 0.11), anxiety (R = 0.11), depression (R = 0.26) and FABQ 2 score (R = 0.23). Disability was poorly correlated with scores on the FABQ 1 (R = 0.45) and FABQ 2 (R = 0.3), anxiety (R = 0.39) and depression (R = 0.47) and not correlated with pain (R = 0.25). Dramatization is the only way to cope with pain and was correlated with pain (R = 0.34), scores on the FABQ 1 (R = 0.47) and FABQ 2 (R = 0.43), disability (R = 0.38), anxiety (R = 0.44) and depression (R = 0.46). The use of prayer was poorly correlated with FABQ 1 score (R = 0.37) and anxiety (R = 0.30). Distraction was poorly correlated with scores on the FABQ 1 (R = 0.40) and FABQ 2 (R = 0.30). No strategy was correlated with duration of pain and stopping work. DISCUSSION-CONCLUSION: All the correlation assessments with psychobehaviour factors are weak. So evaluating each of these parameters will be interesting. Duration of pain was not correlated with increasing pain or change in behavioural strategy. Psychobehavioural factors are more invalidating than pain. More study is needed to assess psychobehavioural therapies in patients with low back pain. PMID- 16716443 TI - Temporal trends in the age-specific prevalence of diabetes and diabetic retinopathy in older persons: Population-based survey findings. AB - PURPOSE: To compare the age-specific prevalence of diabetes and diabetic retinopathy (DR) over 6 years. METHODS: The Blue Mountains Eye Study (BMES) examined 3654 residents (82.4% response) aged >49 years in BMES I (1992-1994). Survivors (n = 2335) and newly eligible residents (n = 1174) were examined in BMES II (1997-2000). Diabetes was defined by history or fasting plasma glucose > or =7.0 mmol/L. DR was graded from 6-field retinal photographs using the Airlie House Classification. Age-specific prevalence in 5-year intervals was compared, as samples of each age group were independent. RESULTS: The overall diabetes prevalence increased from 7.8 to 9.9% (p = 0.002) while the age-specific prevalence increased in age groups 60-74 years, over 6 years. A slightly higher prevalence of mild levels of DR (p = 0.018) but lower prevalence of moderate severe levels of DR (p = 0.049) was evident in BMES II compared to BMES I. Factors significantly associated with DR prevalence were diabetes duration > or =20 years (BMES I, OR 7.6 [2.9-20.5]; BMES II, OR 6.1 [2.5-15.4]) and blood glucose level (BMES I, OR 1.1 [1.0-1.2]; BMES II, OR 1.2 [1.1-1.3] per mmol/L increase). CONCLUSIONS: An increased prevalence of diabetes, but decreased prevalence of severe levels of DR, was documented in two survey samples from the same region over 6 years. PMID- 16716445 TI - A real-time QRS detection method based on moving-averaging incorporating with wavelet denoising. AB - In this paper, a simple moving average-based computing method for real-time QRS detection is proposed. In addition, for signal preprocessing our detection algorithm also incorporates a wavelet-based denoising procedure to effectively reduce the noise level for electrocardiogram (ECG) data. The overall computational structure of the proposed algorithm allows the QRS detection to be performed and implemented in real-time with high time- and memory-efficiency. Algorithm performance was evaluated against the MIT-BIH Arrhythmia Database. The numerical results indicated that the novel algorithm finally achieved about 99.5% of the detection rate for the standard database, and also, it could function reliably even under the condition of poor signal quality in the measured ECG data. PMID- 16716444 TI - Glucose levels in the normal range predict incident diabetes in families with premature coronary heart disease. AB - PURPOSE: Little is known about excess risk of incident diabetes conferred by fasting plasma glucose (FPG) within the normal range (<5.6 mmol/l) for high risk families. METHODS: Healthy 30-59 year old non-diabetic siblings (N = 542) of index cases with documented premature coronary disease were followed prospectively for type 2 diabetes. RESULTS: During 8.7+/-3 years of follow-up, incident diabetes was identified in 7.8%. Rates were incremental with baseline non-diabetes FPG thresholds of 5.0, 5.6, 6.1, and 6.7 mmol/l (p for trend < 0.0001). FPG was the strongest predictor of incident diabetes even across levels within the normal range. The multivariable adjusted relative risk was 14.9 (95% CI = 3.4-65.2) at FPG thresholds > or =5.0 mmol/l versus FPG <5.0 mmol/l. The maximal diagnostic efficiency for FPG was 5.50 mmol/l; with sensitivity and specificity 0.782. All FPG thresholds in the normal range between 5.0 and 5.6 mmol/l showed efficiency levels >0.74. The overall area under the ROC curve predicting incident diabetes for normal and prediabetes ranges of FPG was 0.867. CONCLUSION: Higher FPG levels within the designated "normal" range in high risk families are a potent independent risk factor for type 2 diabetes and may serve as a sentinel to trigger primary preventive interventions. PMID- 16716448 TI - Decades of TREE. PMID- 16716446 TI - Mismatch repair system (MMR) status correlates with response and survival in non small cell lung cancer (NSCLC) patients. AB - Pre-clinical data suggested a relationship between inactivation of hMLH1 and hMSH2 and resistance to drugs like cisplatin and carboplatin, but not oxaliplatin. We then hypothesised that NSCLC showing loss of expression of the mismatch repair system (MMR), could be refractory to cisplatin-based, but not to oxaliplatin-based chemotherapy. Immunohistochemical expression of hMLH1 and hMSH2 was analysed on tumour samples from 93 advanced NSCLC, receiving chemotherapy with either cisplatin or oxaliplatin in combination with gemcitabine. Patients showing loss of hMLH1 or hMSH2 expression in > or = 50% of tumour cells were deemed MMR-negative (Group A), whereas cases with a normal hMLH1 or hMSH2 expression in > 50% of the tumour cells were defined MMR-positive (Group B). No differences in the response and progression rate were found in the whole patients population and in the gemcitabine/cisplatin group for both hMLH1 and hMSH2. In the gemcitabine/oxaliplatin group response rate was 38% and 0% (p=0.04) for patients with or without loss of hMSH2 expression. Median survival according to MMR status in Groups A and B, respectively was: 17 months versus 9 months for hMLH1 (p=0.031) and 10 months versus 9 months for hMSH2 (p=0.8330). Both the difference in response rate and in median survival observed according to MMR status seem to confirm what has been suggested by preclinical studies. PMID- 16716447 TI - Cisplatin plus etoposide chemotherapy followed by thoracic irradiation and paclitaxel plus cisplatin consolidation therapy for patients with limited stage small cell lung carcinoma. AB - PURPOSE: To evaluate the efficacy and tolerance of a cisplatin plus etoposide regimen followed by thoracic radiotherapy (TRT) and paclitaxel plus cisplatin consolidation chemotherapy in patients with limited stage small cell lung cancer (SCLC). PATIENTS AND METHODS: Thirty-nine patients with limited SCLC were enrolled onto this study. Patients received three courses of cisplatin 75 mg/m2 i.v., day 1 and etoposide 100 mg/m2 i.v., days 1-3 (EP regimen), followed by TRT (45-56 Gy administered in 15 fractions), and three courses of paclitaxel 175 mg/m2 i.v., day 1 and cisplatin, as previously, on day 2 (PP regimen); cycles were repeated every 21 days. RESULTS: All patients were evaluable for toxicity and 34 for response. The overall response rate was 67% (CR: 26%; PR: 41%; intention-to-treat analysis) (95% CI: 53.0-84.2%). After a median follow-up period of 15 months, the median survival time was 15 months, the median time to tumor progression 8.3 months and the 1-year survival rate 53.8%. Grade 3/4 neutropenia occurred in 39% and 36% of patients receiving EP and PP regimens, respectively. The incidence of febrile neutropenia was 5% and 3% for EP and PP regimens, respectively. Other hematologic and non-hematologic toxicities were mild, with the exception of esophagitis occurring in 36% of patients during and/or immediately after radiotherapy. CONCLUSION: Consolidation therapy with PP after sequential EP and thoracic radiotherapy is feasible and well-tolerated; however, the efficacy results are comparable with those previously obtained in the same patients' population using a combination of EP and TRT. PMID- 16716449 TI - Competitive sorption and diffusion of chromate and sulphate in a flow system with goethite in gel beads. AB - Column experiments and model simulations were employed to evaluate the processes involved in multicomponent solute transport in a system with heterogeneous flow. Column experiments were performed with goethite embedded in polyacrylamide gel beads. The gel forms an immobile water region that can be accessed by diffusion. A two-region transport model with diffusion into spheres was combined with a surface complexation model to predict reactive transport in the goethite-gel bead system. Chromate and sulphate breakthrough curves were measured in a set of transport experiments, along with corresponding changes in the pH of the effluent. Sorption and transport of sulphate and chromate in separate columns were predicted from independently measured sorption parameters. The model overestimated the pH changes in the effluent, possibly because of proton buffering by the polyacrylamide gel. The effect of competitive sorption on transport was examined in experiments with both anions present. The model predicted the effect of competition very well in a system initially equilibrated with sulphate, followed by infiltration with chromate. However, when sulphate was infiltrated after equilibration with chromate, chromate desorption and sulphate adsorption were clearly overestimated by the transport model. The exchange between the more strongly bound chromate and the sulphate added subsequently may be too slow to cause a substantial chromate peak in the effluent. This suggests that the local equilibrium assumption was not applicable in this case. PMID- 16716450 TI - Modelling radionuclide transport for time varying flow in a channel network. AB - Water flowrates and flow directions may change over time in the subsurface for a number of reasons. In fractured rocks flow takes place in channels within fractures. Solutes are carried by the advective flow. In addition, solutes may diffuse in and out of stagnant waters in the rock matrix and other stagnant water regions. Sorbing species may sorb on fracture surfaces and on the micropore surfaces in the rock matrix. We present a method by which solute particles can be traced in flowing water undergoing changes in flowrate and direction in a complex channel network where the solutes can also interact with the rock by diffusion in the rock matrix. The novelty of this paper is handling of diffusion in the rock matrix under transient flow conditions. The diffusive processes are stochastic and it is not possible to follow a particle deterministically. The method therefore utilises the properties of a probability distribution function for a tracer moving in a fracture where matrix diffusion is active. The method is incorporated in a three dimensional channel network model. Particle tracking is used to trace out a multitude of flowpaths, each of which consists of a large number of channels within fractures. Along each channel the aperture and velocity as well as the matrix sorption properties can vary. An efficient method is presented whereby a particle can be followed along the variable property flowpath. For stationary flow conditions and a network of channels with advective flow and matrix diffusion, a simple analytical solution for the residence time distribution along each pathway can be used. Only two parameter groups need to be integrated along each path. For transient flow conditions, a time stepping procedure that incorporates a stochastic Monte-Carlo like method to follow the particles along the paths when flow conditions change is used. The method is fast and an example is used for illustrative purposes. It is exemplified by a case where land rises due to glacial rebound. It is shown that the effects of changing flowrates and directions can be considerable and that the diffusive migration in the matrix can have a dominating effect on the results. PMID- 16716451 TI - Alternative oxidase regulation in roots of Vigna unguiculata cultivars differing in drought/salt tolerance. AB - The alternative oxidase (Aox) was studied at different levels (transcript, protein and capacity) in response to an osmotic shock applied to roots of cowpea (Vigna unguiculata). Two cultivars of V. unguiculata were used, Vita 3 and Vita 5, tolerant and sensitive to drought/saline stress respectively. The seedlings (17-day-old) were grown in hydroponic conditions and submitted to NaCl (100 and 200 mM) or 200.67 g L(-1) PEG 6000 (iso-osmotic condition to 100 mM NaCl). The VuAox1 and VuAox2a mRNA were not detected in either cultivar under all tested conditions while the VuAox2b gene was differently expressed. In the tolerant cultivar (Vita 3), the expression of VuAox2b gene was stimulated by an osmotic stress induced by PEG which was associated with a higher amount and capacity of the Aox protein. In the same cultivar, this gene was under-expressed in salt stress conditions with poor effect on the protein level. In the sensitive cultivar (Vita 5), the transcript level of the VuAox2b was unchanged in response to PEG treatment, even though the protein and the capacity tended to increase. Upon salt stress, the VuAox2b gene was over-expressed. At 100mM NaCl, this VuAox2b gene over-expression led to a higher amount and capacity of Aox. This effect was reduced at 200 mM NaCl. Overall, these results suggest complex mechanisms (transcriptional, translational and post-translational) for Aox regulation in response to osmotic stress. PMID- 16716452 TI - The symptomless leaf infection with grapevine leafroll associated virus 3 in grown in vitro plants as a simple model system for investigation of viral effects on photosynthesis. AB - The photosynthetic changes evaluated by oxygen evolution, chlorophyll fluorescence, photoacoustics, and delayed fluorescence (DF) were studied in leaves of grown in vitro for 8 weeks grapevine plants (Vitis vinifera) infected by grapevine leafroll-associated virus 3 (GLRaV-3). The infected leaves were characterized during the viral infection without visible disease symptoms. The symptomless infection led to a decrease in plant biomass. The non-photochemical fluorescence quenching, qN, declined, whereas the photochemical quenching, qP, and the Chl a/b ratio were not significantly affected. Photoacoustic and oxygen evolution measurements showed that the energy storage and oxygen evolution rate decreased in the infected leaves. Enhanced alternative electron sinks during the symptomless viral infection were also estimated. The changes in fluorescence and DF temperature curves demonstrated an enhanced stability of the thylakoid membranes in the infected leaves. This effect was clearly expressed at high actinic light intensities. The viral infected in vitro grown grapevine plants were used in the present study as a simplified model system that allow to avoid the involvement of different environmental factors that could interfere with the GLRaV infection and the virus-grapevine interactions. Thus, the 'pure' impact of the viral infection on photosynthesis could be investigated. PMID- 16716453 TI - Expression and localization of calcium-dependent protein kinase isoforms in chickpea. AB - Calcium-dependent protein kinases (CPKs) play important roles in multiple signal transduction pathways but the precise role of individual CPK is largely unknown. We isolated two cDNAs encoding two CPK isoforms (Cicer arietinum CPKs-CaCPK1 and CaCPK2) of chickpea. Their expression in various organs and in response to various phytohormones, and dehydration, high salt stress and fungal spore in excised leaves as well as localization in leaf and stem tissues were analyzed in this study. CaCPK1 protein and its activity were ubiquitous in all tissues examined. In contrast, CaCPK2 transcript, CaCPK2 protein and its activity were almost undetectable in flowers and fruits. Both CaCPK1 and CaCPK2 transcripts and proteins were abundant in roots but in minor quantities in leaves and stems. Of the three phytohormones tested, viz. indole-3-acetic acid (IAA), gibberellin (GA(3)) and benzyladenine (BA), only BA increased both CaCPK1 and CaCPK2 transcripts, proteins and their activities. GA(3) induced accumulation of CaCPK2 transcript and protein but CaCPK1 remained unaffected. The expression of CaCPK1 and CaCPK2 in leaves was enhanced in response to high salt stress. Treatments with Aspergillus sp. spores increased expression of CaCPK1 in chickpea leaf tissue but had no effect on CaCPK2. Excised leaves subjected to dehydration showed increase in CaCPK2 expression but not in CaCPK1. Both isoforms were located in the plasma membrane (PM) and chloroplast membrane of leaf mesophyll cells as well as in the PM of stem xylem parenchyma cells. These results suggest specific roles for CaCPK isoforms in phytohormone/defense/stress signaling pathways. PMID- 16716454 TI - The CART gene: structure and regulation. AB - CART peptides are important neuropeptides that are involved in a variety of physiologic processes. The regulation of the CART gene is critical since peptides are regulated and secreted in response to specific stimuli. CART mRNA must also be controlled in order to respond to specific stimuli such as psychostimulant drugs and leptin. The regulation of the CART gene is central to maintaining homeostasis of peptide production. The 5' upstream region of the CART gene contains powerful regulatory elements that must be involved in transcriptional regulation via different signaling pathways. This review touches on several aspects related to CART gene regulation such as: (i) CART genomic structure, (ii) stimuli that alter CART mRNA levels, (iii) promoter characterization, (iv) role of the cAMP/PKA/CREB signal transduction pathway, and (v) role of the CART 5' and 3' ends in CART mRNA regulation. The goal of this review is to present current data so as to encourage further work in the field of CART gene regulation. PMID- 16716455 TI - Nutrient-induced alpha-amylase and protease activity is regulated by crustacean cardioactive peptide (CCAP) in the cockroach midgut. AB - The midgut plays a major role in digestion and absorption of nutrients in insects, and contains endocrine cells throughout the epithelial layer that express neuropeptides, including crustacean cardioactive peptide (CCAP). In the present study, we demonstrate regulation of digestive enzyme activities by CCAP in response to nutrient ingestion in the cockroach, Periplaneta americana. The midgut of the cockroach exhibits maximal alpha-amylase and protease activities 3 h after intake of either starch or casein, but not of non-nutrients. Similar time dependent responses of CCAP expression in midgut endocrine cells were observed after feeding starch and casein, but not after non-nutrients. We also show that incubation of the dissected midgut with CCAP leads to an increase in alpha amylase and protease activity in a time-dependent manner, with the maximal activity at 2 h. Taken together, our data indicate the existence of an inducible mechanism where endocrine cells in the midgut are stimulated to synthesize and secrete CCAP by nutrients, and CCAP then up-regulates the activity of digestive enzymes. PMID- 16716456 TI - Effect of alpha-melanotropin hormone on serum levels of luteinizing hormone and progesterone in experimental rat autoimmune oophoritis. AB - We studied the effect of alpha-melanotropin hormone (alpha-MSH) on experimental autoimmune oophoritis (EAO), an inflammatory process induced in female rats. During proestrus, serum levels of LH and progesterone in rats with EAO were higher than those of control rats. However, administration of alpha-MSH to these rats decreased the levels of LH. Similarly, in the following diestrus, rats with EAO had high levels of LH but treatment with alpha-MSH decreased the levels to diestrus 2 control values. Treatment with alpha-MSH also reduced the LH levels of control rats in diestrus 2 compared to untreated controls. However, alpha-MSH treatment had no effect on progesterone levels of either control or rats with EAO. Thus, although alpha-MSH induced notable changes in levels of LH, this decrease was unable to block the illness. PMID- 16716457 TI - The anti-nociceptive effect of BmK AS, a scorpion active polypeptide, and the possible mechanism on specifically modulating voltage-gated Na+ currents in primary afferent neurons. AB - In the present study, we investigated the anti-nociceptive effect and the underlying mechanism of BmK AS, an active peptide purified from scorpion Buthus martensi Karsch. The results showed that BmK AS can significantly relieve formalin-induced two-phase spontaneous flinching response and carrageenan-induced mechanical hyperalgesia. Using the whole-cell patch-clamp recording, exposure of acutely isolated sensory neurons to 500 nM BmK AS produced a one-fold decrease in the number of action potentials (APs) evoked by a ramp of depolarizing current. To investigate the mechanism of action of BmK AS, isolated membrane current and Ca2+ influx on rat primary sensory neurons were examined. BmK AS produced insignificant effect on voltage-dependent I(K) and KCl or caffeine-induced Ca2+ influx, but caused remarkable suppressive effect on tetrodotoxin-resistant (TTX R) and tetrodotoxin-sensitive (TTX-S) I(Na). Further experiments showed that BmK AS reduced the peak TTX-R and TTX-S Na+ conductance in a dose-dependent manner, prompted the voltage-dependent activation, and caused a negative shift of the steady-state inactivation of TTX-R and TTX-S I(Na). Thus, the present results indicate the anti-nociceptive response of BmK AS may be ascribed to its specific modulation of voltage-gated Na+ channels of sensory neurons. PMID- 16716458 TI - Synthesis, DNA binding and cytotoxicity of new pyrazole emodin derivatives. AB - A series of new anthrapyrazoles were derived from emodin by attaching various cationic alkyl amino side chains onto a pyrazole ring which had been incorporated into the anthraquinone chromophore. Compared with emodin, the derivatives had significantly higher DNA binding affinity based on interaction with calf thymus DNA, and much more potent cytotoxicity against different tumor cells. The derivatives with a mono-cationic alkyl side chain exhibited the highest DNA binding affinity and cytotoxicity. PMID- 16716459 TI - [Rhabdomyolysis induced by fenofibrate monotherapy]. AB - INTRODUCTION: Rhabdomyolysis with fibrate have been reported when fibrate are associated with statin or during renal insufficiency or hypothyroidism. CASE RECORD: We describe one patient with diabetes mellitus treated by fenofibrate monotherapy since several years; 48 h after gliclazide therapy was introduced, rhabdomyolysis occurred. DISCUSSION: Responsibilities of deshydratation and / or drug interaction with gliclazide. are discussed. PMID- 16716460 TI - Use of rubella seroepidemiological data for assessment of previous vaccination policy and for decision making in response to epidemics in Israel. AB - We examined the prevalence of rubella antibodies in a representative sample of the Israeli population. Three thousand three hundred and twenty-six sera collected during 1997 and 1998, from an age-stratified general population sample were tested for specific IgG antibodies against rubella. The sero-positivity rates to rubella were higher among females as compared with males (89.1% versus 82.3%, respectively (p < 0.001). This difference was the result of much lower sero-positivity rates among males in the age group 13-17, with the lowest value (56.3%) among subjects aged 16. Male subjects of this age group were in 2000 the target of an outbreak of rubella among 18-19-year old male recruits of the Israel Defense Force. The data of this study served to assess previous exposure to the wild virus or vaccine strains, to identify pockets of low level of immunity and contributed to decision making in response to the onset of a rubella outbreak. PMID- 16716461 TI - Prevalence of influenza vaccination and associated factors among community dwelling Hong Kong residents of age 65 or above. AB - A total of 877 Hong Kong Chinese community-dwelling elderly were telephone interviewed. Of them, 63.2% had heard of influenza vaccination (IV), 31.2% had ever had IV (of which 93.3% did so in the last 6 months) and 51.4% were inclined toward receiving free IV in the coming year. Multivariate analyses showed that older age, perceptions of side effects, perceptions of effectiveness duration, and knowing where to get vaccinated were significantly associated with having ever been vaccinated. These variables are related to the Health Belief Model. The prevalence of IV appeared to be increasing as a result of the recent SARS epidemic. PMID- 16716463 TI - A comparison of the soil migration and plant uptake of radioactive chlorine and iodine from contaminated groundwater. AB - A 6-month soil column experiment was conducted to compare the upward migration and plant uptake of radiochlorine and radioiodine from shallow, near-surface contaminated water tables. Both fixed and fluctuating water tables were studied. After 6 months, (36)Cl activity concentrations were relatively uniform throughout the soil profile apart from an accumulation at the soil surface, which was especially marked under a fluctuating water table scenario. In contrast, (125)I (a surrogate for (129)I) tended to accumulate at the boundary between the anoxic conditions at the base of the column and the oxic conditions above, due to its redox-dependent sorption behaviour. The uptake of (36)Cl by perennial ryegrass was much greater than that of (125)I due to its greater migration into the rooting zone and its ready availability in soil solution. In the context of radioactive waste disposal, where these radionuclides may potentially be released into groundwater, (36)Cl would be expected to present a greater potential for contamination of the biosphere than (129)I. PMID- 16716462 TI - Enhanced antigen-specific antibody production following polyplex-based DNA vaccination via the intradermal route in mice. AB - DNA vaccination is an attractive approach with various advantages over conventional vaccination. The present study was undertaken to examine whether polyplex-based DNA vaccination could be used to modulate immune responses by plasmid DNA (pDNA). Methylated bovine serum albumin (mBSA) was used as a model of a cationic macromolecular carrier of pDNA encoding obalbumin (OVA) and the effects of polyplex formation of pDNA with mBSA on the antigen-specific immune responses were examined. Anti-OVA IgG antibody production was significantly increased following intradermal immunization with the polyplex compared with naked pDNA, although the induction of cytotoxic T lymphocyte activity was lowered by polyplex formation. We also demonstrated that the disposition and gene expression of pDNA following intradermal injection could be manipulated by polyplex formation. Intriguingly, we also found that the migration of dendritic cells to the injected site could be induced by polyplex formation probably due to a high level of tumor necrosis factor alpha production from the keratinocytes treated with mBSA/pDNA complexeses. Thus, the present study has demonstrated that the immune responses could be biased towards a Th2-type response by polyplex based DNA vaccination through manipulation of not only pDNA disposition but also dendritic cell migration. PMID- 16716464 TI - Use of a surgeons gown to drape an operating chair. PMID- 16716465 TI - Survey on the perioperative use of TNFalpha inhibitors in rheumatoid hand surgery. PMID- 16716466 TI - Arthoscopic diagnosis of partial scapholunate ligament tears as a cause of radial sided wrist pain in patients with inconclusive x-ray and MRI findings. AB - To clarify the pathology of radial-sided wrist pain with inconclusive X-ray and MRI findings, we performed arthroscopic examinations of 11 wrists in 10 patients. Physical examination and various image investigations could not identify the cause of the pain. Arthroscopy revealed partial to complete tears of the scapho lunate interosseous ligament and synovitis and/or chondral bone defects at the scaphotrapezio-trapezoidal joint in all 11 wrists. Surgical procedures consisted of eight simple synovectomies, two ligament reconstructions and one percutaneous pinning. Pain relief was achieved in 10 wrists. One wrist which had a simple synovectomy did not recover, so underwent secondary scaphotrapezio-trapezoidal fusion. In conclusion, we found that various degrees of scapholunate interosseous ligament tear and scaphotrapezio-trapezoidal joint osteoarthritis were the main causes of radial-sided wrist pain with inconclusive X-ray and simple MRI findings. PMID- 16716468 TI - Pronation contracture of the forearm due to iatrogenic scar formation of the distal membranous part of the forearm interosseous membrane. AB - A case of successful treatment of pronation contracture of the forearm due to iatrogenic scar formation in the distal membranous part of the interosseous membrane of the forearm is presented and the management of this problem is discussed. PMID- 16716469 TI - Diagnostic strategy for suspected scaphoid fractures in the presence of other fractures in the carpal region. AB - Bone scintigraphy will identify up to 25% of occult scaphoid bone fractures after negative scaphoid X-rays. Consequently, it deserves a place in the diagnostic process of suspected scaphoid fractures. However, the role of bone scintigraphy is less clear if scaphoid X-rays show other fractures in the carpal region. We analysed 111 consecutive patients with a suspected scaphoid fracture on physical examination. Scaphoid X-rays revealed 61 fractures. Fifty-five patients had scaphoid fractures only and six patients had other fractures in the carpal region but no scaphoid fracture. In 50 cases, no bone injury was seen on these X-rays. In three out of the six patients with other fractures in the carpal region, bone scintigraphy revealed four occult concomitant fractures: one scaphoid, one scaphoid and trapezial and one capitate fracture. In conclusion, bone scintigraphy is required when scaphoid X-rays do not confirm a suspected scaphoid fracture, even in the presence of other fractures in the carpal region. PMID- 16716470 TI - Re: the anatomical basis for reverse first to fifth dorsal metacarpal arterial flaps. Omokawa et. al. PMID- 16716472 TI - Metacarpal shaft fractures of the fingers: treatment with interosseous loop wire fixation and immediate postoperative finger mobilisation in a wrist splint. AB - It is widely believed that interosseous loop wire fixation of metacarpal shaft fractures alone is not rigid enough to allow immediate postoperative mobilisation. In this report, the author reviews the results in 36 cases of metacarpal shaft fractures of the fingers treated by interosseous loop wire fixation and immediate postoperative mobilisation of all finger joints. The study included 30 males and six females, with a mean age of 31 (range 12-52) years. The fracture pattern was transverse in 26 and oblique, or spiral, in the remaining 10 patients. Following loop wire fixation, the wrist was immobilised using a volar plaster splint for 3 weeks but all finger joints were mobilised immediately after surgery. After a mean follow-up of 8 weeks, all patients, except two, had regained full range of motion of the fingers and no complications were noted. It was concluded that interosseous loop wire fixation of metacarpal shaft fractures is rigid enough to allow immediate postoperative finger mobilisation in a wrist splint and achieve good functional results. PMID- 16716473 TI - Mechanical strength of mediopatellar plica--the influence of its fiber content. AB - BACKGROUND: The fibrous mediopatellar plica can cause high contact pressure on the adjacent articular cartilage and lead to its degeneration. The purpose of this study was to investigate the biomechanical properties of the plica and to correlate this with the plica's fiber content and patients' ages. METHODS: An experimental study on the tensile strength of the mediopatellar plica was conducted using high precision micro-force tensile tests. These tests were undertaken on plica specimens taken from 50 knees of patients with different ages. The force-deflection curves resulting from these tests were recorded and transferred to stress-stain curves to obtain the Young's moduli of these specimens. In addition, pathological tissue dyeing tests were used to assess the fiber content ratio of each specimen. The relationship of the Young's moduli of these specimens with the severity of their pathologic change was also evaluated. FINDINGS: The Young's modulus of the plica was found to be ranging from 10 to 110 MPa. It has positive correlation with patient's age. The relationship between the fiber content ratio and Young's modulus can be fitted properly using a quadratic regression model. The Young's modulus of the plica was also positively correlated with the severity of its pathologic change. INTERPRETATION: The test results indicated that as patients get older, the fiber content of the mediopatellar plica and the Young's modulus of the plica will increase accordingly. We also demonstrated that the Young's modulus of the medial plica was positively correlated with the severity of the plica lesion. PMID- 16716474 TI - A comparison of kinetic gait parameters for 3-13 year olds. AB - BACKGROUND: Normative gait data is essential for diagnosing and treating abnormal gait patterns. The examination of the onset of adult-like kinetic gait patterns in children has generated inconsistent results. The purpose of this study was to identify age-related differences in kinematic and kinetic gait parameters across children aged 3-13 years old. METHODS: A motion capture system and three force plates were employed to compute sagittal joint angles and joint kinetics during walking and compare results between children aged 3-4 years (n=13), 5-6 years (n=10), 7-8 years (n=12), and 9-13 years (n=12). Anthropometric data was estimated using a mathematical model (elliptical cylinder method). Peak flexion and extension joint angles and moments, and peak concentric and eccentric joint powers were analyzed using multivariate analyses of variance. FINDINGS: For most of the variables examined, similar results were obtained across age groups. Reduced peak hip flexion moments and knee extension moments were observed in the 3-4 year olds compared to the oldest group of walkers. Compared to the 9-13 year olds, reduced ankle joint moments and power were observed in most age groups. INTERPRETATION: The results suggest that adult-like kinetic patterns for the hip and knee were attained by 5 years of age. However, for the ankle joint, adult like patterns are not achieved until nine years of age or older. These findings stress the importance of using age-matched normative data for clinical gait analysis. PMID- 16716475 TI - Diagnosis of megaloblastic anaemias. AB - There are a large number of causes of megaloblastic anaemia. The most frequent are disorders resulting in vitamin B(12) or folate deficiency. The diagnostic process often consists first of establishing the presence of B(12) or folate deficiency and then of determining the cause of deficiency. The blood count, blood film, serum B(12) assay, and red cell and serum folate assays are the primary investigations. Other useful investigations include serum/plasma methylmalonic acid (MMA), plasma total homocysteine (tHCYS) and serum holo transcobalamin II assays. All currently used tests have limitations regarding specificity or sensitivity or both and the metabolite assays are not widely available. An understanding of these limitations is essential in formulating any diagnostic strategy. The wide use of serum B(12) and metabolite assays has resulted in the increasingly early diagnosis of B(12) deficiency, often in patients without B(12)-related symptoms (subclinical deficiency). Food cobalamin malabsorption is the most frequent cause of a low serum B(12). At least 25% of low serum B(12) levels are not associated with elevated metabolite levels and may not indicate B(12) deficiency. Some of these are caused by partial deficiency of transcobalamine I. PMID- 16716476 TI - The environmental risk assessment of human pharmaceuticals in the overall EU regulatory affairs process. AB - The approval of new human pharmaceutical products in the EU requires an assessment of potential environmental risks related to the use by patients, besides the evaluation of the human safety, efficacy, and quality evaluation. The current guidance by the European Medicines Agency (EMEA), describing the process of the environmental risk assessment for human drugs, covers a two-tiered assessment programme with a modelling step for an environmental exposure scenario and a subsequent step of environmental fate and effects testing. The following paper describes ways how the requirements of the environmental risk assessment can be sensibly incorporated in the overall approval process of a pharmaceutical product including the risk/benefit analysis for the patient. The resources for environmental testing and assessment programmes can be employed economically, if the pharmacological, toxicological, and pharmacokinetic information obtained during the development programme of a human pharmaceutical is used to develop substance-specific test programmes and to evaluate the environmental risk assessment taking into account the pharmacodynamic properties and the use pattern by patients. Finally, we suggest that the environmental risk evaluation process as part of drug approvals should adhere to a focussed assessment strategy considering existing knowledge and the therapeutic needs. PMID- 16716478 TI - How does social comparison within a self-help group influence adjustment to chronic illness? A longitudinal study. AB - Despite the growing popularity of self-help groups for people with chronic illness, there has been surprisingly little research into how these may support adjustment to illness. This study investigated the role that social comparison, occurring within a self-help group, may play in adjustment to chronic illness. A model of adjustment based on control process theory and response shift theory was tested to determine whether social comparisons predicted adjustment after controlling for the catalyst for adjustment (disease severity) and antecedents (demographic and psychological factors). A sample of 301 people with Meniere's disease who were members of the Meniere's Society UK completed questionnaires at baseline and 10-month follow-up assessing adjustment, defined for this study as functional and goal-oriented quality of life. At baseline, they also completed measures of the predictor variables i.e. the antecedents (age, sex, living circumstances, duration of self-help group membership, self-esteem, optimism and perceived control over illness), the catalyst (severity of vertigo, tinnitus, hearing loss and fullness in the ear) and mechanisms of social comparison within the self-help group. The social comparison variables included the extent to which self-help group resources were used, and whether reading about other members' experiences induced positive or negative feelings. Cross-sectional results showed that positive social comparison was indeed associated with better adjustment after controlling for all the other baseline variables, while negative social comparison was associated with worse adjustment. However, greater levels of social comparison at baseline were associated with a deteriorating quality of life over the 10-month follow-up period. Alternative explanations for these findings are discussed. PMID- 16716477 TI - Distrust, social fragmentation and adolescents' health in the outer city: Beirut and beyond. AB - This study examined the association between place and components of social capital among adolescents living in three impoverished communities outside of Beirut, the capital city of Lebanon. We utilized a unique data set that includes a wide range of social capital items to assess the association between place, social capital and self rated health. The analyses were based on data collected by trained interviewers during the Spring of 2003 from a stratified random sample of 1294 adolescents aged 13-19 years, drawn from a sampling frame constructed from area maps and detailed household listing of the target population. Descriptive statistics and odds ratios from logistic regression models were used to assess the strength of associations between (1) the social capital items and community of residence and (2) social capital, community and self-rated health controlling for age, sex and income. Findings show that distrust and social fragmentation were generally prevalent among adolescents living in impoverished suburban communities. Even though social networks, especially the presence of family and relatives, were strong, instrumental social exchange was relatively scarce. Social capital items varied significantly by community, and the stock of social capital in one of the three communities surveyed-the Palestinian refugee camp-was quite distinctive. Findings from logistic regression models showed that social capital as measured by a simple index is strongly associated with self rated health status, controlling for community, age, sex and income. Some implications of the findings are discussed. PMID- 16716479 TI - Oxidation of apolipoprotein B-containing lipoproteins and serum paraoxonase/arylesterase activities in major depressive disorder. AB - Major depressive disorder (MDD) is blaimed to play a role in the onset of coronary artery disease (CAD). The aim of the present study was to investigate serum paraoxonase/arylesterase activities and oxidation of apolipoprotein B containing lipoproteins in patients with MDD. Oxidation of lipoproteins plays an important role in atherogenesis and the enzyme paraoxonase, has been shown to prevent lipoprotein oxidation. Furthermore, low paraoxonase activity was suggested to predict CAD. Eighty-six patients who fully met the fourth Diagnostic and Statistical Manual of Mental Disorders criteria for MDD and 36 healthy control subjects were included in the study. Serum paraoxonase and arylesterase activities were determined spectrophotometrically. Malondialdehyde (MDA) levels of apolipoprotein B-containing lipoproteins were determined before (basal) and after incubation with copper-sulphate, that yielded basal- and Delta-MDA values, respectively. Serum paraoxonase/arylesterase activities were significantly reduced in the post-treatment group compared with the pre-treatment group. Basal MDA (MDA) level was significantly higher in the MDD group compared with the control group. Delta-MDA level of the severe MDD group was significantly higher than that of the control group. There was a positive correlation between the oxidizability of apolipoprotein B-containing lipoproteins and the severity of the disease. Total cholesterol, HDL-cholesterol, LDL-cholesterol, apolipoprotein B levels were significantly higher and apolipoprotein AI levels were significantly lower in the MDD group compared with those of the control group. The findings of the present study suggest that: 1) antidepressant treatment might reduce serum paraoxonase activity/mass; 2) oxidation and oxidizability of apolipoprotein B containing lipoproteins seem to be increased in MDD. PMID- 16716480 TI - Human erythrocytes are affected in vitro by extracts of Ugni molinae leaves. AB - Ugni molinae Turcz, also known as "Murtilla", is a plant that grows in the south of Chile. Infusions of their leaves have long been used in traditional native herbal medicine. The chemical composition of the leaves indicates the presence of polyphenols, which have antioxidant properties. In order to evaluate the mechanisms of their antioxidant properties and the toxicity of the aqueous extracts of leaves, the extracts were induced to interact with human red cells, their isolated unsealed membranes (IUM) and large unilamellar vesicles (LUV) of dimyristoylphosphatidyltidylcholine (DMPC), representative of phospholipid classes located in the outer monolayer of the erythrocyte membrane. Scanning electron microscopy (SEM) observations indicated that the extracts achieved a significant alteration in the shape of the erythrocytes as they changed their discoid shape to echinocytes. According to the bilayer couple hypothesis, the shape change indicates that the polyphenols were located in the outer moiety of the red cell membrane. This conclusion was confirmed by the fluorescence experiments performed in IUM and DMPC LUV. In fact, the extracts produced slight initial increases followed by sharp decreases at higher concentrations in the anisotropy and general polarization parameters. These results imply that the extracts induced structural perturbations in the acyl chain and polar group packing arrangements of the erythrocyte IUM and DMPC LUV lipid bilayers: first ordering and afterwards disordering them as the extract concentration increased. PMID- 16716481 TI - Effects of tetrabromobisphenol A, brominated flame retardant, in ICR mice after prenatal and postnatal exposure. AB - Tetrabromobisphenol A (TBBPA), brominated flame retardant, is produced in the largest amounts globally for use in plastics or building materials. TBBPA has been detected in sediment, air at the dismantling plant or human serum samples. In the present study, we examined the effects of prenatal and postnatal exposure to TBBPA in mice. TBBPA (99.1% pure) in diet was administered to pregnant ICR mice at doses of 0% (control), 0.01%, 0.1% or 1.0% from gestational day 0 to weaning at postnatal day 27. The average daily food intake and body weight of dams showed no significant differences between the control and treated groups. There were no dose-related effects on reproductive data. Serum concentrations of total-cholesterol and liver weights of treated dams and offspring were higher than those of the control mice. Histological findings in treated dams or offspring showed the increase of focal necrosis of hepatocytes and inflammatory cell infiltration in the liver, and increase of dilation or atrophy of renal tubules and cyst in the kidney. TBBPA was developed as a new, safe class of flame retardant and was not highly toxic. However, the present data suggested that TBBPA caused a lipid metabolic disorder and hepatic or kidney lesion, under these conditions. PMID- 16716482 TI - Glutathione S-transferase polymorphisms and ochratoxin A toxicity in primary human urothelial cells. AB - The mycotoxin ochratoxin A (OTA) is a worldwide contaminant of human food. OTA is genotoxic, immunotoxic, teratogenic and carcinogenic in rodents and can cause nephropathy in pigs. High amounts of OTA can cause nephropathy in humans. Moreover, evidence has been accumulated that OTA is a genotoxic carcinogen. Nevertheless, the mechanism that leads to OTA toxicity has not been fully resolved and it is discussed if a bioactivation of OTA is necessary or not. In this study the genotoxicity of OTA was investigated in primary human urothelial cells by means of alkaline single cell gel electrophoresis (Comet assay). Primary cultured human urothelial cells derived from tissue specimens of urological patients were incubated with 100 microM OTA for 3 h. In contrast to recently published results in MDCK cell lines, the cell cultures showed great interindividual differences in the extent of DNA damage. To evaluate these great interindividual differences the influence of the genotype of the isoenzymes of glutathione S-transferase (GST), namely GSTT1, GSTM1 and GSTP1 on the genotoxic potential of OTA was examined. The genotypes of these polymorphic enzymes were determined by polymerase chain reaction (PCR) and the distributions of the genotypes were correlated with the extent of DNA damage. We found associations between the genotypes of the polymorphic GST isoenzymes and the extent of DNA damage between subgroups with and without OTA-related DNA damage. From these results we conclude that genetic predisposition has the potential to influence OTA genotoxicity. PMID- 16716483 TI - Metabolomic, proteomic and biophysical analyses of Arabidopsis thaliana cells exposed to a caesium stress. Influence of potassium supply. AB - The incorporation and localisation of 133Cs in a plant cellular model and the metabolic response induced were analysed as a function of external K concentration using a multidisciplinary approach. Sucrose-fed photosynthetic Arabidopsis thaliana suspension cells, grown in a K-containing or K-depleted medium, were submitted to a 1 mM Cs stress. Cell growth, strongly diminished in absence of K, was not influenced by Cs. In contrast, the chlorophyll content, affected by a Cs stress superposed to K depletion, did not vary under the sole K depletion. The uptake of Cs was monitored in vivo using 133Cs NMR spectroscopy while the final K and Cs concentrations were determined using atomic absorption spectrometry. Cs absorption rate and final concentration increased in a K depleted external medium; in vivo NMR revealed that intracellular Cs was distributed in two kinds of compartment. Synchrotron X-ray fluorescence microscopy indicated that one could be the chloroplasts. In parallel, the cellular response to the Cs stress was analysed using proteomic and metabolic profiling. Proteins up- and down-regulated in response to Cs, in presence of K+ or not, were analysed by 2D gel electrophoresis and identified by mass spectrometry. No salient feature was detected excepting the overexpression of antioxidant enzymes, a common response of Arabidopsis cells stressed whether by Cs or by K-depletion. 13C and 31P NMR analysis of acid extracts showed that the metabolome impact of the Cs stress was also a function of the K nutrition. These analyses suggested that sugar metabolism and glycolytic fluxes were affected in a way depending upon the medium content in K+. Metabolic flux measurements using 13C labelling would be an elegant way to pursue on this line. Using our experimental system, a progressively stronger Cs stress might point out other specific responses elicited by Cs. PMID- 16716484 TI - The ribosome's response to codon-anticodon mismatches. AB - The ribosome is a molecular machine that synthesizes polypeptides from aminoacyl tRNAs according to the sequence of the mRNA template. Codon reading by the anticodon of tRNA is controlled by a network of ribosome contacts that are specific for each position of the codon-anticodon duplex and involve A-minor RNA interactions. Rapid and accurate tRNA selection is accomplished by switching the conformation of the decoding site between accepting and rejecting mode, regardless of the thermodynamic stability of the respective codon-anticodon complexes or their interactions at the decoding site. The forward reactions are particularly sensitive to mismatches and determine the variations in the extent of misreading of near-cognate codons, both during initial selection and proofreading. This review emphasizes the progress made in understanding the mechanisms that determine recognition and selection of tRNA by the translational machinery. PMID- 16716485 TI - Autoregulation of Oct-1 gene expression is mediated by two octa-sites in alternative promoter. AB - Transcription of the oct-1 gene is regulated by two alternative promoters: U promoter and L promoter located upstream of the exons 1U and 1L, respectively. The L promoter contains two octamer sequences of opposite orientation: proximal (ATTTGCAT) and distal (ATGCAAAT), showing high affinity toward the Oct proteins. Binding of the Oct-1 protein to the octa-sites located in the L promoter region has been confirmed in footprinting experiments. Dual luciferase assay using wild type and mutated promoters have indicated that mutations in the proximal octa site resulted in significant transcription enhancement both in myeloma cell line NS/0 and in fibroblast cell line 3T3 (about twofold and fivefold, respectively), whereas mutations in the distal site decreased the promoter activity (about 10% and 40%, respectively). Mutations in both octa-sites enhanced the effect and increased transcription to about fourfold in myeloma cell line NS/0 and about sixfold in fibroblast cell line 3T3. These results demonstrate that transcription of the oct-1 gene may be autoregulated by two octa-sites within the L promoter. Different function and interactive tissue-specific effect of distal and proximal octamer sequences can be suggested. PMID- 16716486 TI - Inhibitory circuits in the thalamus and hippocampus--an appraisal after 40 years. AB - After scientific successes in the study of synaptic activation and inhibition of motoneurons and unraveling mechanisms underlying presynaptic inhibition, Sir John Eccles was interested in studying synaptic mechanisms governing the activity of neurons in the brain stem, cerebellum, and various cortical areas. In this new arena, his group discovered several principles, which have later been shown to generalize across brain structures and have substantial functional significance. Among these were the first identification and location of inhibitory synapses in the cerebral cortex and recurrent inhibitory systems in the hippocampus, cerebellum, and thalamus. PMID- 16716487 TI - Role of the AP-1 transcription factor c-Jun in developing, adult and injured brain. AB - The transcription factor activator protein-1 (AP-1) consists of a variety of dimers composed of members of the Jun and Fos families of proteins. However, it is the up-regulation of c-jun that is a particularly common event in the developing, adult as well as in injured nervous system that serves as a model of transcriptional control of brain function. In view of the long list of excellent in depth overviews on the different members of the Jun family and associated molecules, the primary focus of the current paper is to focus on c-Jun specifically and discuss the evidence on the involvement of this transcription factor in ischaemia and stroke, in seizures, during learning and memory, or following axonal injury and during successful regeneration. Functional studies employing in vivo strategies using gene deletion, targeted expression of dominant negative isoforms and pharmacological inhibitors all suggest a bipotential role of c-Jun, in mediating neurodegeneration and cell death, as well as in plasticity and repair. Phosphorylation of c-Jun, and the activation of its upstream kinases (Jun N-terminal Kinase (JNK) 1-3, JNK kinases (JNKK)) is required in many but not in all forms of these events, with only a partial overlap of the Jun-, JNK- or JNKK(n)-dependent functions. Moreover, a better understanding of the non overlapping roles could considerably increase the potential of pharmacological agents to improve neurological outcome following trauma, neonatal encephalopathy and stroke, or neurodegenerative disease. PMID- 16716489 TI - Diversity of potassium channels in neuronal dendrites. AB - Complex computations in the nervous system begin with electrical signals generated in single neurons. Such signals include action potentials mediated by the opening of voltage-dependent ion channels, and synaptic potentials arising from neurotransmitter receptor activation. The amplitude, waveform, and propagation of action potentials and synaptic potentials influence cellular signaling in profound ways, and are largely determined by activities of ion channels in the cell membrane. The location and properties of ion channels therefore play critical roles in shaping electrical signaling in the neuron, which is the foundation for more complex computations at network levels. This review summarizes what we know about the great diversity of K(+) channels found in neuronal dendrites, the subcellular compartment where synaptic signals integrate and where various forms of plasticity occur. Specifically, we discuss the molecular identity, the distribution, kinase modulation, biophysical properties, and functional roles of a variety of K(+) channels including voltage gated, calcium-activated, and ligand-gated/G-protein coupled K(+) channels. One emerging theme from recent literature is the recognition that K(+) channels are powerful regulators of the function of dendrites. A second theme indicates that this K(+) channel regulation depends on their unique subcellular distribution. In particular, the mechanisms underlying the establishment and maintenance of non uniform distributions of ion channels are beginning to be understood in greater detail. An especially intriguing aspect of above mechanisms is that they are achieved through protein kinase phosphorylation and may thus be activity dependent. In parts of this review, we choose to focus on CA1 pyramidal neurons of the rodent hippocampus and the K(+) channels in their dendrites. Being one of the best-characterized cell types in the nervous system, the CA1 pyramidal neuron has long been studied as a prototypic neuron from which general rules of neuronal computation and synaptic plasticity emerge. A great deal of what we know about dendritic K(+) channels comes from studies on CA1 pyramidal neurons. Where available, we also include up-to-date findings on dendritic K(+) channels in other cell types. PMID- 16716488 TI - Spinal reflexes, mechanisms and concepts: from Eccles to Lundberg and beyond. AB - This review focuses on investigations by Sir John Eccles and co-workers in Canberra, AUS in the 1950s, in which they used intracellular recordings to unravel the organization of neuronal networks in the cat spinal cord. Five classical spinal reflexes are emphasized: recurrent inhibition of motoneurons via motor axon collaterals and Renshaw cells, pathways from muscle spindles and Golgi tendon organs, presynaptic inhibition, and the flexor reflex. To set the scene for these major achievements I first provide a brief account of the understanding of the spinal cord in "reflex" and "voluntary" motor activities from the beginning of the 20th century. Next, subsequent work is reviewed on the convergence on spinal interneurons from segmental sensory afferents and descending motor pathways, much of which was performed and inspired by Anders Lundberg's group in Gothenburg, SWE. This work was the keystone for new hypotheses on the role of spinal circuits in normal motor control. Such hypotheses were later tested under more natural conditions; either by recording directly from interneurons in reduced animal preparations or by use of indirect non-invasive techniques in humans performing normal movements. Some of this latter work is also reviewed. These developments would not have been possible without the preceding work on spinal reflexes by Eccles and Lundberg. Finally, there is discussion of how Eccles' work on spinal reflexes remains central (1) as new techniques are introduced on direct recording from interneurons in behaving animals; (2) in experiments on plastic neuronal changes in relation to motor learning and neurorehabilitation; (3) in experiments on transgenic animals uncovering aspects of human pathophysiology; and (4) in evaluating the function of genetically identified classes of neurons in studies on the development of the spinal cord. PMID- 16716490 TI - Cytokine concentrations during the first days of life. AB - OBJECTIVE: To evaluate the cytokine concentration patterns during the first 5 days of life by measuring serum concentrations of type-1 cytokines, like interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) and type-2 cytokines, like IL-4, as well as the receptors of IL-2 (sIL-2R) and IL-4 (sIL-4R) during the early neonatal period. SUBJECTS AND METHODS: Forty-two healthy term neonates were included in the study. Cytokine concentrations were measured in umbilical cord, in the 1st and 5th day after birth and compared with those in serum of 30 healthy adults. RESULTS: IL-2 concentrations presented a decrease trend from umbilical cord to 5th day, while sIL-2R showed a significant elevation from umbilical cord to 5th day after birth. IL-4 concentrations did not differ significantly among umbilical cord, the 1st and the 5th day, while the sIL-4R showed the highest values in the 1st day after birth. Both IL-4 and sIL-4R concentrations in neonatal samples were elevated compared to adults. IFN-gamma concentrations increased significantly from umbilical cord to 5th day of life. CONCLUSION: Our findings indicate a dysregulation among IL-2, IL-4 and IFN-gamma concentrations during the 1st day after birth, favoring a more precocious expression of IL-2 and IL-4 against IFN-gamma that seems to be ameliorated in the end of the 1st week of life. PMID- 16716491 TI - Non-closure versus closure of peritoneum during cesarean section: a randomized study. AB - BACKGROUND: Adhesion bands are reported to be among the most common complications of gynecological surgery procedures. The aim of this study was to compare the frequency of post-surgical adhesion among patients with closed or open peritoneal repair in the subsequent cesarean sections. METHODS: A prospective randomized trial was performed on 45 patients who underwent cesarean section during a 5-year period. Among these patients, 24 patients were operated on by closure of the peritoneum and 21 patients were operated on by the non-closure method. Overall, 31 patients had a second pregnancy and cesarean section within the time of the study. These patients were evaluated to determine if they had any adhesion between the omentum and abdominal wall or uterus or between the uterus and abdominal wall or rectus muscle. RESULTS: The two groups were similar to each other with regard to the causes of cesarean section. Seven cases of adhesions were diagnosed during subsequent cesarean sections among 13 patients (54%) with peritoneal non-closure, compared with 3 in 18 women (15%) with peritoneal closure (relative risk: 3.2; 95% confidence interval: 1.0-10.2). CONCLUSION: Closure of the peritoneum increases the operating time, but may decrease the risk of adhesions. The results of this study suggest that, during cesarean section, suture of the peritoneum may be a better option than leaving it unsutured. PMID- 16716493 TI - Selected biologically relevant ions at the air/water interface: a comparative molecular dynamics study. AB - Interfacial behavior of selected biologically and technologically relevant ions is studied using molecular dynamics simulations employing polarizable potentials. Propensities of choline, tetraalkylammonium (TAA), and sodium cations, and sulfate and chloride anions for the air/water interface are analyzed by means of density profiles. Affinity of TAA ions for the interface increases with their increasing hydrophobicity. Tetramethylammonium favors bulk solvation, whereas cations with propyl and butyl chains behave as surfactants. The choice of counter anions has only a weak effect on the behavior of these cations. For choline, sodium, chloride and sulfate, the behavior at the air/water interface was compared to the results of our recent study on the segregation of these ions at protein surfaces. No analogy between these two interfaces in terms of ion segregation is found. PMID- 16716492 TI - The stability and characteristics of a DNA Holliday junction switch. AB - A Holliday junction (HJ) consists of four DNA double helices, with a branch point discontinuity at the intersection of the component strands. At low ionic strength, the HJ adopts an open conformation, with four widely spaced arms, primarily due to strong electrostatic repulsion between the phosphate groups on the backbones. At high ionic strength, screening of this repulsion induces a switch to a more compact (closed) junction conformation. Fluorescent labelling with dyes placed on the HJ arms allows this conformational switch to be detected optically using fluorescence resonance energy transfer (FRET), producing a sensitive fluorescent output of the switch state. This paper presents a systematic and quantitative survey of the switch characteristics of such a labelled HJ. A short HJ (arm length 8 bp) is shown to be prone to dissociation at low switching ion concentration, whereas an HJ of arm length 12 bp is shown to be stable over all switching ion concentrations studied. The switching characteristics of this HJ have been systematically and quantitatively studied for a variety of switching ions, by measuring the required ion concentration, the sharpness of the switching transition and the fluorescent output intensity of the open and closed states. This stable HJ is shown to have favourable switch characteristics for a number of inorganic switching ions, making it a promising candidate for use in nanoscale biomolecular switch devices. PMID- 16716494 TI - The benefits and costs of reducing emissions from the electricity sector. AB - Recent federal policy proposals to reduce emissions of sulfur dioxide (SO(2)), nitrogen oxides (NO(x)), and mercury from the US electricity sector promise important improvements in air quality and reductions in acid deposition. The cost of achieving these reductions depends on the form and stringency of the regulation. In this research, we analyze the economic benefits and costs of the US Environmental Protection Agency's (EPA's) Clean Air Interstate Rule (CAIR) as characterized in the supplemental rule proposed in June 2004, and the Clean Air Mercury Rule (CAMR) as proposed in February 2004. The assessment integrates a model of the electricity sector, two models of atmospheric transport of air pollutants, and a model of environmental and public health endpoints affected by pollution. We model explicitly the emissions of SO(2), NO(x), mercury and carbon dioxide (CO(2)) and the effects of changes in emissions of SO(2) and NO(x) on environmental and public health. The manner in which mercury emissions are regulated will have important implications not only for the cost of the regulation, but also for emission levels for SO(2) and NO(x) and where those emissions are located. We find the economic benefits of CAIR and CAMR are far greater than the costs. Recent estimates of benefits of reductions in mercury and acidification indicate that our model captures the lion's share of quantifiable benefits. We also find that the EPA would have been justified on economic grounds in pursuing additional SO(2) emissions reductions beyond the requirements of CAIR. PMID- 16716495 TI - Surgical treatment of Peyronie's disease: a critical analysis. AB - OBJECTIVE: The present paper reviews surgical treatment alternatives for patients with Peyronie's disease using knowledge obtained from the contemporary literature. METHODS: : All aspects of surgical treatment for Peyronie's disease were examined on the basis of MEDLINE database researches. RESULTS: Surgical treatment should be delayed until the acute inflammatory phase has resolved and should be considered in patients with deformity that impairs sexual function. Currently, surgical treatment alternatives are reconstructive surgery by either lengthening the concave side (incision and grafting) or shortening the convex side (Nesbit procedure or plication) of the penis, and implantation of penile prosthesis with or without incision of the plaque. PD patients with good erectile capacity are candidates for reconstructive surgery. Meanwhile, implantation of penile prosthesis with or without remodeling should be considered for patients without adequate erectile capacity. CONCLUSIONS: The aim of the surgical treatment in Peyronie's disease is to correct the deformity while preserving or improving erectile capacity of the penis. Appropriate treatment options should be individualized according to the patients' expectations and erectile capacity. PMID- 16716496 TI - Levels of polychlorinated biphenyls (PCBs) in transformer oils from Korea. AB - Using Korean waste official method, polychlorinated biphenyls (PCBs) were analyzed from 33 transformer oils collected under National Institute of Environmental Research (NIER) in Republic of Korea (ROK) and their emission rate in Korea was estimated. Practical quantification limits analyzed by this method were in the ranges of 0.02 and 0.05 mg L(-1), and recovery of samples was in the ranges of 77-97%. The analytical results showed that PCBs in the transformer oils were identified as Aroclor 1242, 1254 or 1260. Twenty-nine samples were determined to contain PCBs of >0.05 mg L(-1) and among these samples, nine samples contained to have >2 mg kg(-1) which is limit of Waste Control Act (WCA) with regard to PCBs in transformer oil from Korea. The concentrations of PCBs in transformer oils were between N.D. (not detected) and 48.33 mg kg(-1). PCBs concentrations did not exceed 50 mg kg(-1) which is generally accepted limit for the definition of waste in POPs Guideline of Basel Convention. PMID- 16716498 TI - Kinetic and isotherm studies of cadmium adsorption on manganese nodule residue. AB - The adsorption equilibrium and kinetics studies of cadmium (Cd) ions from aqueous solutions on manganese nodule residue were carried out by considering the influence of various parameters, such as contact time, solution pH and initial metal concentration in solution, temperature and adsorbent quantity. The adsorption of Cd increased with an increase in the concentrations of this metal in solution. Presence of manganese and iron content in manganese nodule residue (MNR) played a significant role in Cd(II) ions removal. The linear forms of the Langmuir and Freundlich equations were utilized for experiments with metal concentrations of 200 mg/L for Cd(II) as functions of solution pH (2.0-6.0). First-order rate equation and pseudo second-order rate equations were applied to study adsorption kinetics. Mass transfer study was also done to know the reaction rate. Thermodynamic parameters, such as standard Gibb's free energy (deltaG(o)), standard enthalpy (deltaH(o)) and standard entropy (deltaS(o)), were also evaluated by Van't Hoff equation. Thus, adsorption of Cd on this adsorbent was found to be spontaneous and exothermic thermodynamically. The adsorption capacity for Cd was found to be 19.8 mg/g of MNR. Under the optimised conditions, cadmium level was brought down from 100 mg/L to Cd less than detection limits and from 200 to 2 mg/L. Thus, the wastewater after cadmium removal could be safely disposed off on to land or sewage. Finally, the metal loaded adsorbent was subjected to desorption using different mineral acids and leaching by using toxicity characteristic leaching procedure (TCLP), synthetic precipitation leaching procedure (SPLP) tests for its further treatment or its safe disposal. PMID- 16716497 TI - Sono-sorption as a new method for the removal of lead ion from aqueous solution. AB - Sorption of Pb(II) as a toxic heavy metal ion, onto styrene-divinylbenzene co polymer resin in the presence of ultrasound (sono-sorption) and in its absence (classical method) is presented in this work. Batch experiments were conducted to study the effect of the main parameters such as the amount of sorbent, concentration of sorbate, contact time, intensity of ultrasound and temperature on this sorption process. The equilibrium between liquid and solid phase was described by Langmuir model better than the Freundlich model. The time to reach equilibrium especially in the presence of ultrasound was very fast. This indicates that the resin mentioned has a high potential for the sorption of this pollutant ion. The isotherm constants were obtained under several different conditions. These constants were used in the calculation of the thermodynamic parameters of sorption. According to the results, the kinetics of sorption was in agreement with both pseudo-first-order and first-order reversible models. PMID- 16716499 TI - Auto-ignition and upper explosion limit of rich propane-air mixtures at elevated pressures. AB - The auto-ignition limits of propane-air mixtures at elevated pressures up to 15 bar and for concentrations from 10 mol% up to 70 mol% are investigated. The experiments are performed in a closed spherical vessel with a volume of 8 dm3. The auto-ignition temperatures decrease from 300 degrees C to 250 degrees C when increasing the pressure from 1 bar to 14.5 bar. It is shown that the fuel concentration most sensitive to auto-ignition depends on initial pressure. A second series of experiments investigates the upper flammability limit of propane air mixtures at initial temperatures up to 250 degrees C and pressures up to 30 bar near the auto-ignition area. Finally the propane auto-oxidation is modelled using several detailed kinetic reaction mechanisms and these numerical calculations are compared with the experimental results. PMID- 16716500 TI - Distribution of extractable fractions of heavy metals in sludge during the wastewater treatment process. AB - Sludge samples were collected from different treatment steps of Gaobeidian wastewater treatment plant (WWTP) of Beijing City, PR China, to investigate the distributions of total and chemical fractions of Fe, Mn, Ni, Cu, Zn, Cr, Pb, and Mo in different sludges. The highest total concentrations were found for Fe, Mn, Pb, and Mo in digested sludge (DS), Ni and Cr in thickened sludge (TS), Zn in dewatering sludge (DWS), and Cu in active sludge (AS). The lowest concentrations were observed in AS, except for Cu in TS. Significant differences of total metal concentration were observed between AS and TS (or DS), suggesting that sludge thickening and digesting treatments significantly influenced the total metal concentrations. Fe, Cu, Ni, Cr, Mo, and Pb distributed principally in the residual fraction in all sludges, while Zn and Mn presented in a highly available fraction. For same metal in different sludges, the portion of easily mobile fraction decreased significantly along the wastewater treatment process, and metals in AS presented in the highest available fraction. Organic matter contents, TN, and TP of sludges exhibited a significant positive correlation with the concentrations of exchangeable and reducible fraction of Pb, Mo, Cr, Cu, and Fe, while sludge pH demonstrated significant negative correlations with the concentrations of these metals. PMID- 16716501 TI - The investigation on cationic exchange capacity of zeolites: the use as selective ion trappers in the electrokinetic soil technique. AB - The cation exchange capacity (CEC) of porous zeolites allows to adsorb in the framework cavities the cations as pollutant heavy metal ions. We investigate the CEC behaviour of different zeolites in different experimental conditions; in solution where the ion's mobility is spontaneous and free and in the electrokinetic system where the ion's mobility is driven by the electric field. The aim of this study is to investigate if the CEC is an useful property to create a special interface region of zeolites, that if placed in the electrokinetic cell, just before the cathode, could allow to capture and concentrate the heavy metallic ions, during their migrating process. The zeolite 13X investigated in the electrokinetic proofs, retains a good high ions adsorption, even if quite smaller than the relevant free solution condition and well acts as confined trap for the heavy metal ions. In fact no trace of metallic deposition are present on the electrode's surface. PMID- 16716502 TI - Barrier and operational risk analysis of hydrocarbon releases (BORA-Release). Part II: Results from a case study. AB - This paper presents results from a case study carried out on an offshore oil and gas production platform with the purpose to apply and test BORA-Release, a method for barrier and operational risk analysis of hydrocarbon releases. A description of the BORA-Release method is given in Part I of the paper. BORA-Release is applied to express the platform specific hydrocarbon release frequencies for three release scenarios for selected systems and activities on the platform. The case study demonstrated that the BORA-Release method is a useful tool for analysing the effect on the release frequency of safety barriers introduced to prevent hydrocarbon releases, and to study the effect on the barrier performance of platform specific conditions of technical, human, operational, and organisational risk influencing factors (RIFs). BORA-Release may also be used to analyse the effect on the release frequency of risk reducing measures. PMID- 16716503 TI - Photodegradation of orange I in the heterogeneous iron oxide-oxalate complex system under UVA irradiation. AB - To understand the photodegradation of azo dyes in natural aquatic environment, a novel photo-Fenton-like system, the heterogeneous iron oxide-oxalate complex system was set up with the existence of iron oxides and oxalate. Five iron oxides, including gamma-FeOOH, IO-250, IO-320, IO-420 and IO-520, were prepared and their adsorption capacity was investigated in the dark. The results showed that the saturated adsorption amount (gamma(max)) was ranked the order of IO-250 > IO-320 > gamma-FeOOH > IO-420 > IO-520 and the adsorption equilibrium constant (Ka) followed the order of IO-250 > IO-520 > gamma-FeOOH > IO-420 > IO-320. The effect of initial pH value, the initial concentrations of oxalate and orange I on the photodegradation of orange I were also investigated in different iron oxide oxalate systems. The results showed that the photodegradation of orange I under UVA irradiation could be enhanced greatly in the presence of oxalate. And the optimal oxalate concentrations (C(ox)0) for gamma-FeOOH, IO-250, IO-320, IO-420 and IO-520 were 1.8, 1.6, 3.5, 3.0 and 0.8 mM, respectively. The photodegradation of orange I in the presence of optimal C(ox)0 was ranked as the order of gamma FeOOH > IO-250 > IO-320 > IO-420 > IO-520. The optimal range of initial pH was at about 3-4. The first-order kinetic constant for the degradation of orange I decreased with the increase in the initial concentration of orange I. Furthermore, the variation of pH, the concentrations of Fe3+ and Fe2+ during the photoreaction were also strongly dependent on the C(ox)0 and iron oxides. PMID- 16716504 TI - Glass-ceramic materials from electric arc furnace dust. AB - Electric arc furnace dust (EAFD) was vitrified with SiO2, Na2CO3 and CaCO3 powders in an electric furnace at ambient atmosphere. Vitreous products were transformed into glass-ceramic materials by two-stage heat treatment, at temperatures determined by differential thermal analysis. Both vitreous and glass ceramic materials were chemically stable. Wollastonite (CaSiO3) was separated from the parent matrix as the dominant crystalline phase, verified by X-ray diffraction analysis and energy dispersive spectrometry. Transmission electron microscopy revealed that wollastonite crystallizes mainly in its monoclinic form. Knoop microhardness was measured with the static indentation test method in all initial vitreous products and the microhardness values were in the region of 5.0 5.5 GPa. Devitrification resulted in glass-ceramic materials with microhardness values strongly dependent on the morphology and orientation of the separated crystal phase. PMID- 16716505 TI - Preparation and application of organo-modified zeolitic material in the removal of chromates and iodides. AB - The removal of chromates and iodides from aqueous solutions by organo-modified tuffs from the Pentolofos area (Thrace, Greece) was investigated using (51)Cr- and (131)I-labelled solutions and gamma-ray spectroscopy. The zeolitic material was modified by hexadecyltrimethyl-ammonium bromide (HDTMA-Br) and octadecyltrimethyl-ammonium bromide (ODTMA-Br) and characterized by scanning electron microscopy, FT-IR spectrometry and zeta potential measurements. Both experimental study and modelling indicated that both organo-zeolitic sorbents have a bigger affinity for iodide than for chromate. The chromium uptake did not seem to be influenced by the type of modifier but showed, as expected, a dependence on the solution pH. The maximum sorption capacity (2.27 mg/g) of Cr(VI) was achieved for the solution of initial pH 4. On the other hand, the HDTMA-modified tuff showed a lower sorption affinity for iodides than did the ODTMA-modified one (3.37 and 4.02 mg/g, respectively). PMID- 16716506 TI - Expression of GABAA receptor beta2/3 subunits in the rat major pelvic ganglion. AB - Several pharmacological and physiological studies have suggested that GABA(A) receptors (GABA(A) Rs) may exist in the rat major pelvic ganglion (MPG), a large coalescent pelvic ganglion that contains both sympathetic and parasympathetic components which innervates pelvic organs. However, the presence of GABA(A) R in the MPG has never been demonstrated directly by morphological studies. In the present study, we used immunohistochemistry to demonstrate the existence of GABA(A) R beta2/3 subunits for the first time in the rat MPG. We also analyzed the neurochemical properties of MPG neurons expressing GABA(A) R beta2/3 subunits. GABA(A) R beta2/3-immunoreactive (-IR) neurons occupied 27.4+/-7.0% of the whole neuronal population, and many of these (77.6%) were co-localized with tyrosine hydroxylase (TH). Likewise, most (86.5%) of TH-IR neurons were GABA(A) R beta2/3-positive. GABA(A) R beta2/3 subunits were also expressed in a few VIP- or NOS-IR neurons, the cholinergic or non-adrenergic, non-cholinergic (NANC) neurons. These results suggest that GABA(A) Rs are involved in the modulation of most sympathetic, noradrenergic neurons and also a subset of VIP and NOS neurons of the rat MPG. PMID- 16716507 TI - Development of baclofen tolerance in a rat model of chronic spasticity and rigidity. AB - Systemic or spinal treatment with baclofen has been associated with the development of tolerance in patients with chronic spasticity. In the present study, we used a rat model of spinal ischemia-induced spasticity to characterize the development of baclofen tolerance after chronic intrathecal (i.t.) baclofen infusion. Following the induction of spinal ischemia and the development of behavioral spasticity, animals were implanted with i.t. catheters connected to osmotic pumps to continuously infuse baclofen (1.0 microg/0.5 microl/h). Hindleg peripheral muscle resistance (PMR) was measured periodically after initiation of chronic infusion and after bolus i.t. baclofen injection (1.0 microg). Peripheral muscle resistance was significantly decreased at the onset of baclofen infusion, however, after 5-7 days of infusion a progressive return of spasticity was noted, where baseline PMR values returned to preinfusion levels. At the same time, the efficacy of bolus i.t. baclofen treatment also decreased, where after 5 days of baclofen infusion 1.0 microg (i.t.) baclofen only reduced PMR by 10% (compared to 40-50% preinfusion). Baclofen efficacy progressively returned once continuous infusion was stopped. These data demonstrate that transient spinal ischemia leads to the development of spasticity which is sensitive to spinal baclofen. Chronic i.t. infusion leads to a progressive development of tolerance. This model offers potential to study tolerance mechanisms after spinal injury, and aid in drug discovery for use in baclofen-tolerant patients. PMID- 16716508 TI - Mu opioid receptors are extensively co-localized with parvalbumin, but not somatostatin, in the dentate gyrus. AB - In the rat dentate gyrus, mu opioid receptor (MOR) agonists disinhibit principal cells, promoting excitation, but whether MOR protein is differentially distributed to interneuron subtypes is unknown. Here, the distribution of MOR immunoreactivity was semi-quantitatively examined in neurochemically identified interneurons using fluorescence microscopy. We find that MOR- and parvalbumin immunoreactivities are frequently co-localized, while MOR- and somatostatin immunoreactivities are less commonly co-localized. This suggests that MORs are most frequently on interneurons specialized to inhibit granule cell output, and are on a limited number of interneurons that inhibit granule cell distal dendrites. PMID- 16716509 TI - An MEG study into the visual perception of apparent motion in depth. AB - This study investigated (1) the responses associated with the perception of motion in depth induced by a series of scale-changing paradigms, and (2) the locations of the dipoles in the brain elicited by stimuli of motion in depth and in-plane motion. The former was determined using two types of stimulation: real motion and apparent motion in scale (AMS; where two frames with size-changed patterns were presented alternately); and the latter was determined by moving a full-field checkerboard pattern forward or rightward smoothly with the same time course. The results from analyzing magnetoencephalography (MEG) component M160 to differentiate the signals of perception can be summarized as follows: (1) the neurons stimulated by apparent motion (AM) might be similar to those stimulated by a real motion, since there was no statistical difference associated with the signals at M160 and the dipole locations; (2) the perceptional signal of motion in depth seems to be more sensitive when scale-changing information is present; and (3) asymmetrical responses are present in the visual system, with responses being more sensitive to expanding stimuli than to contracting ones, and with the activity being more prominent in the right occipitotemporal area. Overall, this study indicates that the responses evoked by the stimuli causing motion in depth are allocated more to area V3a rather to area V5. PMID- 16716510 TI - Differential time course and intensity of PFC activation for men and women in response to emotional stimuli: a functional near-infrared spectroscopy (fNIRS) study. AB - Using functional near-infrared spectroscopy (fNIRS) we recorded prefrontal cortex (PFC) activation during positive, negative and neutral film clips, based on affective ratings according to their valence and arousal, to assess gender differences in cerebral activation in 15 male and 15 female volunteers. To record PFC activation, five movie clips were presented on a 17-in. TFT screen. The recordings included a pre-stimulus 5-s local baseline and "on" and "off" segments of data, referring to fNIRS Oxy-Hb levels while stimulation (movie clip) was present and during an inter-stimulus blank screen. Our data showed gender differences in the delay period to initial PFC activation and in the course and intensity of activation produced by affective visual stimuli. During the exposure or "on" period of the stimuli we observed more pronounced overshoot and undershoot in men versus women across the range of emotions elicited. This effect was even more pronounced following stimulus cessation ("off" period). The results indicate that gender and the duration of recordings may affect the results of emotional neuroimaging studies. PMID- 16716511 TI - Visual search for biological motion: an event-related potential study. AB - To investigate the neural response to detection of biological motion (BM) surrounded by distractors, event-related potentials (ERPs) were recorded. Scrambled motion with the same velocity vector as the BM but randomized initial starting points was used as the distractor. The number of distractors was varied to control the difficulty of the task. The behavioral data showed that the reaction time increased with the number of distractors. Moreover, the ERP results showed that enhanced negativity was elicited at posterior electrodes contralateral to the location of the BM as with the conventional N2pc-like component, which is related to selection of the target. The increment of reaction time indicated that the attentional process is involved in the detection of BM. Furthermore, the ERP waveforms suggest that, even when detection of the target requires form-from-motion processing, similar neural mechanisms to those involved in conventional visual search tasks were employed here. PMID- 16716512 TI - gamma-Tocopherol attenuates MPTP-induced dopamine loss more efficiently than alpha-tocopherol in mouse brain. AB - Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated mouse has been widely used as a rodent model of Parkinson's disease. In this study, alpha-tocopherol (alphaT) transfer protein knockout (heteromutant type, alpha-TTP((+/-))) mice were used to evaluate the protective effects of alphaT and gamma-tocopherol (gammaT) against MPTP-induced neurotoxicity. The intraperitoneal administration of MPTP to mice induced a decrease in the striatal levels of dopamine (DA) 3 days after the administration in both alpha-TTP((+/-)) and wild-type mice; these mice were fed an alphaT-deficient diet for 3 weeks before the MPTP administration. The DA levels in the alpha-TTP((+/-)) mice, which had been fed a gammaT-fortified diet (0.10 wt.%) for 3 weeks and were administered with MPTP, were recovered to those of the control, whereas there was no significant protective effect of alphaT despite the considerably higher striatal concentration of alphaT than gammaT. The immunohistochemical study also revealed that gammaT exerted a protective effect against neurodegenerative toxicity of MPTP. Collectively, this is the first report showing that the protective effect of gammaT is stronger than that of alphaT against the MPTP-induced damage of dopaminergic neurons in the mouse. PMID- 16716513 TI - Observations of crypt neuron-like cells in the olfactory epithelium of a cartilaginous fish. AB - A new receptor neuron (RN) type was recently described in bony fish olfactory epithelium (OE): the crypt receptor neuron. This name is due to its main feature: the presence, at the apical part, of a deep invagination into which cilia protrude. The presence of this receptor neuron type is well documented in different species of bony fishes but it has never been described in cartilaginous fishes. In this study we demonstrate that crypt neuron-like cells are present in the olfactory epithelium of the elasmobranch Scyliorhinus canicula (Linnaeus, 1758). Histological observations allowed us to detect the presence of a few egg shaped cells, characterized by a crypt like zone; alpha-tubulin immunoreactivity suggested the presence of cilia in the same area; fluorocrome conjugated lectin bindings suggested a distinctive mucus composition inside the presumptive crypt. The possible presence of crypt neuron-like cells in chondrichthyes would represent an interesting common feature between bony and cartilaginous fishes. PMID- 16716514 TI - Modulation of brain apoptosis-related proteins by the opioid antagonist naltrexone in mice. AB - Neuronal loss by apoptosis has been implicated in some neural pathologic disorders. Increasing evidence suggests a neuroprotective effect for opioid antagonists, such as naloxone and naltrexone, in a variety of neural damage experimental models and in the clinic. The purpose of the present study was to analyse the effects of naltrexone on the expression levels of proteins regulating the extrinsic (FasL and Fas) and the mitochondrial (Bcl-2, Bcl-xL, Bad and Bax) apoptotic pathways, as well as the active fragment of the executioner caspase-3 in the mouse brain. Western blotting showed that a single injection of naltrexone (1 mg/kg) induced a down-regulation of the pro-apototic proteins Fas, FasL, Bad and Bax. Our results suggest that naltrexone provides neuronal protection against injuries activating either mitochondrial, or death receptor-apoptotic pathways. PMID- 16716515 TI - Corticotropin releasing factor in urine--a possible biochemical marker of fibromyalgia. Responses to massage and guided relaxation. AB - The purpose of this preliminary study was to evaluate the relationship between a possible biochemical marker of stress, 24-h urinary concentrations of Corticotropin Releasing Factor-Like Immunoreactivity (CRF-LI), and ratings of stress-related symptoms like depression and anxiety, as well as to evaluate pain and emotional reactions in patients with fibromyalgia (FM). Another purpose was to study the effects of massage and guided relaxation, with respect to change in the same variables. Urine sampling and ratings were performed before treatments, after and 1 month after completed treatments. Concentrations of CRF-LI was analysed with radioimmnoassay technique. For the assessment of depression, anxiety and pain the CPRS-A questionnaire was used and for rated pain and emotional reactions the NHP questionnaire was used. The 24-h urinary concentration of the CRF-LI was found to be related to depression, mood and inability to take initiative. After treatment the urinary CRF-LI concentrations and the rated levels of pain and emotional reactions were found to have decreased. In conclusion, the 24-h urinary CRF-LI concentration may be used as a biochemical marker of stress-related symptoms such as depression in patients with FM and possibly also other conditions characterized by chronic pain. Therapies such as massage and guided relaxation may be tried for the amelioration of pain and stress but further studies are required. PMID- 16716516 TI - Testing two accounts of pain underestimation. AB - Two important influences on pain underestimation by health care professionals were investigated by varying specific cues with reference to underestimation of patients' pain: when observers are not allowed to talk to patients and when observers expect social cheating. One hundred and twenty health care professionals watched videotaped facial expressions of pain patients and estimated their pain. The first group only saw the faces, the second group was given patients' self-reports in addition and the last group was given a context cue priming them to expect cheating in addition to faces and patients' ratings. Health care professionals generally underestimated patients' pain, but this varied depending on the cues given. Those viewing the face without patients' ratings underestimated pain to a greater extent than health care professionals provided with patients' ratings. Health care professionals primed to expect cheating underestimated pain as much as those seeing only patients' faces. Therefore, both accounts, verbal report as important but missing cue as well as an alerted cheating detection device, could account for underestimation. PMID- 16716517 TI - Explaining sex differences in chronic musculoskeletal pain in a general population. AB - Many studies report a female predominance in the prevalence of chronic musculoskeletal pain (CMP) but the mechanisms explaining these sex differences are poorly understood. Data from a random postal questionnaire survey in the Dutch general population were used to examine whether sex differences in the prevalences of CMP are due to sex differences in the distribution of known potential risk factors for CMP (exposure model) and/or to the different importance of risk factors for CMP (i.e. show different strength of association) in men and women (vulnerability model). In the present analyses, 909 men and 1178 women aged 25-65 were included. CMP was defined as pain lasting longer than 3 months and was assessed for 10 anatomical locations (neck, shoulder, higher back, elbow, wrist/hand, lower back, hip, knee, ankle, foot). Sex differences in CMP could not be explained by a different distribution of age, educational level, smoking status, overweight, physical activity, and pain catastrophizing. Having no paid job was associated with CMP, explaining part of the sex differences, but its role seems complex. Risk factors with a sex-specific association were: overweight (all pain locations) and older age (lower extremities)--both having only an effect among women--and pain catastrophizing (upper extremities), which was stronger associated with CMP among men than among women. In conclusion, sex differences in prevalence of CMP may partly be explained by sex differences in vulnerability to risk factors for CMP. Future research towards sex-specific identification of risk factors for CMP is warranted. Eventually this may lead to sex-specific prevention and management of CMP. PMID- 16716518 TI - Sensory neuropathy and signs of central sensitization in patients with peripheral arterial disease. AB - Patients with peripheral arterial disease (PAD) may develop a broad range of peripheral nerve dysfunctions including pain and sensory deficiencies due to chronic ischemia mostly involving the lower limbs. To investigate the degree of sensory abnormalities in such patients quantitative sensory testing (QST) might be a useful tool. Forty-five patients and 20 controls were enrolled in the present study and underwent QST according to the protocol of the German Research Network on Neuropathic Pain. PAD was graded according to the Rutherford classification. PAD patients were divided into two groups: 16 patients with critical limb ischemia (severe PAD) and 29 patients with intermittent claudication (moderate PAD). QST revealed impaired cold and warm detection, increased mechanical and vibration detection thresholds, and increased perceptual wind-up on the affected leg (all p<0.001). Paradoxical heat sensation (p<0.05) and dynamic mechanical allodynia (p<0.01) were also observed. Subgroup analysis of patients without diabetes (control n=20, moderate PAD n=21, severe PAD n=8) confirmed most of these findings. In patients with severe PAD, sensory deficits were more pronounced than in patients with moderate PAD and were detected even in the face. These data indicate that QST can detect sensory abnormalities in PAD patients. While the pattern of decreased perception suggests deafferentation for Abeta-, Adelta-, and C-fiber inputs, the presence of allodynia suggests that central sensitization also plays a role in the pain state of PAD patients. Subgroup analysis points towards a PAD-associated peripheral neuropathy independent of diabetes. PMID- 16716519 TI - An atypical case of Trypanosoma cruzi infection in a young English Mastiff. AB - A case of Trypanosoma cruzi infection in a young English Mastiff from Texas is presented. Clinical signs and laboratory findings included subcutaneous edema, lymphadenopathy, weight loss, and hypoalbuminemia. Cytology of a lymph node revealed numerous amastigotes. No trypomastigotes were observed in buffy coat preparation of peripheral blood, and on histologic evaluation, most organs contained numerous interstitial pseudocysts. Initial serology was positive for both T. cruzi and Leishmania, and immunohistochemistry supported a diagnosis of Leishmania. However, additional serology supported a T. cruzi infection, and cultivation of organisms isolated from a lymph node revealed morphology consistent with T. cruzi. In addition, PCR analysis resulted in a 504 bp fragment with 99% homology to a flagellar protein of T. cruzi. Although uncommon, autochthonous cases of both T. cruzi and Leishmania have been reported in the United States. Clinical signs observed with both diseases can show many similarities, cytology may be indistinguishable, as in this case, and serological cross-reactivity is common. This case demonstrates an unusual presentation of T. cruzi and the use of multiple testing strategies to support its diagnosis. PMID- 16716520 TI - Culicoides bottimeri as a vector of Haemoproteus lophortyx to quail in California, USA. AB - Arthropod sampling via periodic direct bird examination and regular light trapping was conducted between June 2000 and October 2002 to survey for potential vectors of Haemoproteus lophortyx to nonnative, captive-raised bobwhite quail (Colinus virginianus) in northern California, USA. Examination of individual bobwhite quail (from 5 weeks of age through adult, total n=76) was conducted on several dates during the transmission period (June-October). No ectoparasites, including hippoboscid flies (reported as Haemoproteus vectors to wild quail in early literature), were collected from the birds. Trapping with ultraviolet light suction traps near the quail revealed nine species of biting midges (Culicoides spp.). Of these, 94% were C. bottimeri, which was abundant near the birds, and 65% of collected C. bottimeri were engorged with blood. C. bottimeri adult activity began in late-April, slightly before the onset of disease in the quail. Activity peaked between July and late-September, coincident with maximum reported transmission, and adult activity ceased by early-November. Nonengorged C. bottimeri had a parity rate of 43.6% overall, suggesting excellent survival for biological transmission of a pathogen like H. lophortyx. A controlled study was done injecting a macerated slurry of pooled, nonengorged, wild-caught C. bottimeri into the peritoneum of 1-day-old bobwhite quail hatchlings held in insect-proof containers. Blood smears 13-19 days later confirmed H. lophortyx infection in zero controls but all insect-injected quail. Biting midges, especially C. bottimeri, transmit H. lophortyx to captive quail and probably are the dominant vector to native California quail (Callipepla californica) as well. PMID- 16716521 TI - Control of Haemonchus contortus in goats with a sustained-release multi-trace element/vitamin ruminal bolus containing copper. AB - The objective of this study was to determine the effectiveness of a sustained release multi-trace element/vitamin ruminal bolus (TEB) containing copper administered to mature does for control of gastrointestinal nematodes (GIN) during summer months and during late gestation. Haemonchus contortus was the predominant nematode during these trials. In Experiment 1, yearling Spanish does were untreated or administered TEB (n=11/group) on Day 0 (August 2005) prior to breeding. In Experiment 2, Spanish and Boer does were untreated or administered TEB 6 weeks before kidding. Fecal egg counts (FEC) and blood packed cell volume (PCV) were determined weekly between Days 0 and 42. In both experiments, FEC were reduced within 7 days in TEB-treated compared with untreated does. PCV was similar between treatment groups (Experiment 1) or tended to be lower in the TEB group (Experiment 2). GIN control did not persist more than 28 days. These studies suggest that TEB may be an effective means of GIN control in mature goats, but additional control measures may be necessary. PMID- 16716522 TI - Complex formation between endogenous toxin bilirubin and polyamidoamine dendrimers: a spectroscopic study. AB - The effects of 4th and 5th generation cationic, neutral and anionic polyamidoamine (PAMAM) dendrimers on bilirubin absorbance and fluorescence were studied. Cationic and neutral PAMAM dendrimers shifted the bilirubin absorption maximum from 435 to 442-455 nm, increased the peak absorbance 1.5-fold, shifted the bilirubin fluorescence excitation and emission maxima, increased the fluorescence emission several-fold and significantly protected bilirubin against photodestruction. Using double fluorescence titration technique allowed to receive such constant of binding and the number of binding centers at 20 degrees C: for PAMAM G4 dendrimer, (2.4+/-1.4) x 10(6) (mol/l)(-1) and 0.07+/-0.012; for PAMAM G4-OH dendrimer, (3.1+/-1.3) x 10(6) (mol/l)(-1) and 0.08+/-0.014; for PAMAM G5 dendrimer, (7.6+/-3.6) x 10(6) (mol/l)(-1) and 0.09+/-0.02; and for PAMAM G5-OH dendrimer, (8.5+/-3.2) x 10(6) (mol/l)(-1) and 0.09+/-0.02. These effects can be explained by the formation of bilirubin-PAMAM dendrimer complexes and the formation of bilirubin monomers from tetramers. The formation of complexes sharply increased bilirubin solubility. We conclude that cationic and neutral PAMAM dendrimers bind bilirubin effectively and suggest that such dendrimers may serve as detoxication agents for hydrophobic endogenous toxins. PMID- 16716523 TI - Habenula lesions alter synaptic plasticity within the fimbria-accumbens pathway in the rat. AB - Both the habenula and the nucleus accumbens, and especially the glutamatergic innervation of the latter from the hippocampus, have been hypothesized to be involved, in different ways, in the pathophysiology of cognitive disturbances in schizophrenia. Lesions of the habenula produce disturbances of memory and attention in experimental animals. As the habenular nuclei have been shown to influence the release of many neurotransmitters, both in the hippocampus and the nucleus accumbens, we examined in this study the effects of bilateral habenula lesions on the plasticity of the fimbria-nucleus accumbens pathway, by means of the long-term depression phenomenon in freely moving rats. Long-term depression, induced within the shell region of the nucleus accumbens by low-frequency stimulation of the fimbria, was exaggerated and showed greater persistence in habenula-lesioned rats compared with sham-operated animals. These results indicate that plasticity in the fimbria-nucleus accumbens pathway is altered by habenula lesions in a way similar to previously-reported effects of stress and the psychosis-provoking agent ketamine. Moreover, they strengthen the views that the habenula belongs to systems, mediating higher cognitive functions, which involve the hippocampus and the nucleus accumbens. Finally, this study suggests that dysfunction of the habenula could contribute to cognitive alterations in diseases such as schizophrenia, where the habenula is reported to exhibit exaggerated calcification. PMID- 16716524 TI - Differential effects of voluntary physical exercise on behavioral and brain derived neurotrophic factor expression deficits in Huntington's disease transgenic mice. AB - Huntington's disease is a fatal neurodegenerative disorder caused by a mutation of the huntingtin gene and involves progressive motor abnormalities (including chorea), cognitive deficits (dementia) as well as psychiatric symptoms. We have previously demonstrated that environmental enrichment slows the onset and progression of Huntington's disease in transgenic mice. Here, we investigated the effects of enhanced physical exercise on disease progression and brain-derived neurotrophic factor expression. Standard-housed Huntington's disease mice developed phenotypic rear-paw clasping by 16 weeks of age, displayed abnormal rearing behavior, deficits in motor co-ordination and of spatial working memory. Huntington's disease mice with access to running wheels exhibited delayed onset of rear-paw clasping, normalized levels of rearing behavior and amelioration of the cognitive deficits. However, in contrast to our previous environmental enrichment studies, there was no rescue of motor coordination deficits in wheel running Huntington's disease mice. An abnormal accumulation of brain-derived neurotrophic factor protein in the frontal cortex of Huntington's disease mice was unaffected by running. Striatal and hippocampal brain-derived neurotrophic factor protein levels were unchanged. Brain-derived neurotrophic factor mRNA levels were reduced in the anterior cortex, striatum and hippocampus of Huntington's disease mice, and only striatal deficits were ameliorated by running. Overall, we show that voluntary physical exercise delays the onset of Huntington's disease and the decline in cognitive ability. In addition, our results reveal that some aspects of hippocampal dependent memory are not entirely reliant on sustained hippocampal brain-derived neurotrophic factor expression. PMID- 16716525 TI - Sensitivity of rat temporalis muscle afferent fibers to peripheral N-methyl-D aspartate receptor activation. AB - The temporalis muscle is a common source of pain in headache and chronic craniofacial pain conditions such as temporomandibular disorders, which have an increased prevalence in women. The characteristics of slowly conducting temporalis afferent fibers have not been investigated. Therefore, the aim of the present study was to examine the characteristics of slowly conducting temporalis muscle afferent fibers and to determine whether these fibers are excited by activation of peripheral N-methyl-D-aspartate receptors. The response properties of a total of 117 temporalis afferent fibers were assessed in male and female rats. A majority of these fibers had high mechanical thresholds and slow conduction velocities (<10 m/s). The mechanical threshold of the temporalis afferent fibers was inversely correlated with afferent conduction velocity, however, no sex-related differences in mechanical threshold were identified. There were also no sex-related differences in N-methyl-D-aspartate-evoked afferent discharge. Indeed, injection of a high concentration (1600 mM) of N methyl-D-aspartate into the temporalis muscle was necessary to evoke significant afferent discharge. Thirty minutes after the initial injection of N-methyl-D aspartate into the temporalis muscle, a second injection of N-methyl-D-aspartate produced a response only about 50% as large as the initial injection. Co injection of ketamine (20 mM) with the second injection of N-methyl-D-aspartate significantly decreased N-methyl-D-aspartate-evoked afferent discharge in both sexes. This concentration of ketamine is greater than that needed to attenuate afferent discharge evoked by injection of glutamate into the masseter muscle. These results suggest that unlike masseter afferent fibers, temporalis afferent fibers are relatively insensitive to peripheral N-methyl-D-aspartate receptor activation. PMID- 16716526 TI - Chronic morphine administration results in tolerance to delta opioid receptor mediated antinociception. AB - Delta opioid receptor agonists produce only a moderate degree of antinociception, possibly reflecting the predominantly intracellular location of delta opioid receptor. However, recent studies suggest that short term morphine pretreatment can increase delta opioid receptor-mediated antinociception by promoting the translocation of delta opioid receptor to the cell surface. Even more striking sensitization has been reported after long term morphine pretreatment and withdrawal in locomotor tests. In the present study we therefore examined the effects of longer term morphine pretreatment and withdrawal on delta opioid receptor-mediated antinociception in the formalin test. Male adult rats were pretreated daily with morphine (10 mg/kg s.c.) or saline for 10 days, and were tested acutely with the delta opioid receptor agonist [D-Ala2,Glu4]-deltorphin (intrathecal) at 0, 7 and 14 days of withdrawal. Unexpectedly, chronic morphine pre-exposure resulted in tolerance to [D-Ala2,Glu4]-deltorphin-induced antinociception, and this occurred at 0 and 7 but not 14 days of morphine withdrawal. Morphine challenge at withdrawal day 7 confirmed the presence of tolerance to the antinociceptive effects of this drug. Chronic morphine pretreatment also resulted in tolerance to the locomotor stimulant effect of [D Ala2,Glu4]-deltorphin (given i.c.v.), contrary to a previous report of sensitization. However, consistent with previous reports, short term (2 day) pretreatment with morphine did result in sensitization to [D-Ala2,Glu4] deltorphin. Subsequent in vitro analysis, using [125I][D-Ala2,Glu4]-deltorphin or guanosine 5'(gamma-35S-thio) triphosphate autoradiography, did not reveal any changes in delta opioid receptor binding or function resulting from chronic morphine pretreatment. In conclusion, chronic morphine pretreatment caused tolerance to delta opioid receptor-mediated behavioral effects with no clear change at the receptor level. PMID- 16716527 TI - Sensory system-predominant distribution of leukotriene A4 hydrolase and its colocalization with calretinin in the mouse nervous system. AB - Leukotriene B4 is a potent lipid mediator, which has been identified as a potent proinflammatory and immunomodulatory compound. Although there has been robust evidence indicating that leukotriene B4 is synthesized in the normal brain, detailed distribution and its functions in the nervous system have been unclear. To obtain insight into the possible neural function of leukotriene B4, we examined the immunohistochemical distribution of leukotriene A4 hydrolase, an enzyme catalyzing the final and committed step in leukotriene B4 biosynthesis, in the mouse nervous system. Immunoreactivity for leukotriene A4 hydrolase showed widespread distribution with preference to the sensory-associated structures; i.e. neurons in the olfactory epithelium and vomeronasal organ, olfactory glomeruli, possibly amacrine cells, neurons in the ganglion cell layer and three bands in the inner plexiform layer of the retina, axons in the optic nerve and tract up to the superior colliculus, inner and outer hair cells and the spiral ganglion cells in the cochlea, vestibulocochlear nerve bundle, spinal trigeminal tract, and lamina II of the spinal cord. Double immunofluorescence staining demonstrated that most of the leukotriene A4-hydrolase-immunopositive neurons coexpressed calretinin, a calcium-binding protein in neurons. The ubiquitous distribution of leukotriene A4 hydrolase was in sharp contrast with the distribution of leukotriene C4 synthase [Shimada A, Satoh M, Chiba Y, Saitoh Y, Kawamura N, Keino H, Hosokawa M, Shimizu T (2005) Highly selective localization of leukotriene C4 synthase in hypothalamic and extrahypothalamic vasopressin systems of mouse brain. Neuroscience 131:683-689] which was confined to the hypothalamic and extrahypothalamic vasopressinergic neurons. These results suggest that leukotriene B4 may exert some neuromodulatory function mainly in the sensory nervous system, in concert with calretinin. PMID- 16716528 TI - Alterations of the expression and activity of midbrain nitric oxide synthase and soluble guanylyl cyclase in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced Parkinsonism in mice. AB - The study was aimed at investigating the expression and the activity of neuronal nitric oxide synthase, and of soluble guanylyl cyclase and phosphodiesterase activities that regulate guanosine 3',5'-cyclic monophosphate level in the midbrain, in a mouse model of PD using 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine injections. Adult male mice of the C57/BL strain were given three i.p. injections of physiological saline or three i.p. injections of 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine solution in physiological saline at 2 h intervals (summary 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine dose: 40 mg/kg), and were killed 3, 7, or 14 days later. mRNA, protein level, and/or activities of neuronal nitric oxide synthase, soluble guanylyl cyclase, phosphodiesterase and guanosine 3',5'-cyclic monophosphate were determined. Immunohistochemistry showed about 75% decrease in the number of tyrosine hydroxylase-positive neurons in the substantia nigra pars compacta. Mice treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine showed increased midbrain guanylyl cyclase and total nitric oxide synthase activities at 3, 7, and 14 days post-treatment. The specific neuronal nitric oxide synthase inhibitor 7 nitroindazole (10 microM) and the specific inducible nitric oxide synthase inhibitor 1400W (10 microM) inhibited the 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine-induced excess in nitric oxide synthase activity by 63-70 and 13-25%, respectively. The increases in total midbrain nitric oxide synthase activity were accompanied by elevated guanosine 3',5'-cyclic monophosphate, enhanced expression of neuronal nitric oxide synthase and of the beta1 subunit of guanylyl cyclase at both mRNA and protein levels that persisted up to the end of the observation period, and by enhanced neuronal nitric oxide synthase and guanylyl cyclase beta1 immunoreactivities in substantia nigra pars compacta 7 and 14 days after the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine treatment. The increases in guanylyl cyclase activity were found to occur exclusively due to increased maximal enzyme activity. No 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine-induced change in phosphodiesterase activity has been detected in any brain region studied. 7-Nitroindazole prevented a significant increase in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced midbrain guanosine 3',5' cyclic monophosphate level and neurodegeneration of dopaminergic neurons. These results raise the possibility that the nitric oxide/guanylyl cyclase/guanosine 3',5'-cyclic monophosphate signaling pathway may play a role in maintaining dopaminergic neurons function in substantia nigra pars compacta. PMID- 16716529 TI - Difference in pre- and post-treatment plasma DHEA levels were significantly and positively correlated with difference in pre- and post-treatment Hamilton depression scores following successful therapy for major depression. AB - OBJECTIVE: Clinical studies of endogenous concentrations of dehydroepiandrosterone (DHEA) and its relation to depression are limited. This study examined whether pre- and post-treatment changes in plasma DHEA levels are correlated with pre- and post-treatment differences in Hamilton depression scores following successful antidepressant therapy for major depression with venlafaxine XR. METHOD: Thirty-four medication-free major depressive outpatients (Hamilton Rating Scale for Depression 17, HAM-D 17 score > or = 17) were treated with antidepressants. At baseline, plasma DHEA levels of all subjects were measured but only those who remitted (HAM-D 17 score < or = 7) before the end of this study had their plasma DHEA levels measured at remission-onset. Blood from subjects was drawn at 0900-1100 h. Depression severity was assessed with the HAM D 17 scale at baseline, and on day 7, 14, 28, 56 and 84. Subjects were administered at minimum 75 mg/day venlafaxine XR until remission onset. RESULTS: Fifteen patients remitted before the end of this study. Plasma DHEA levels decreased from baseline to remission was significant (P=0.017). After controlling for age and gender, pre- and post-treatment difference in Hamilton depression scores and the pre- and post-treatment difference in DHEA concentrations were significantly correlated (P=0.044). CONCLUSION: This preliminary study provides the first clinical evidence identifying that difference in pre- and post treatment plasma DHEA levels were significantly and positively correlated with difference in pre- and post-treatment Hamilton depression scores following successful therapy with venlafaxine XR for major depression in remitters; but non remitters were not examined. It is not known if DHEA levels would show similar or dissimilar changes in non-remitters. PMID- 16716530 TI - Increased cortisol in women with intimate partner violence-related posttraumatic stress disorder. AB - BACKGROUND: Alterations of hypothalamic-pituitary-adrenal (HPA) axis function and sympathetic-adrenal activity have been proposed as key factors in biological models of posttraumatic stress disorder (PTSD). METHODS: We examined neuroendocrine function in female survivors of intimate partner violence (IPV) with lifetime (current or remitted) PTSD (n=29) and in women who were exposed to IPV but never developed PTSD (n=20). Salivary cortisol was collected as a marker of HPA axis function at 1, 4, 9, and 11 h after awakening. Platelet epinephrine and norepinephrine were assayed as markers of sympathetic-adrenal activation. RESULTS: Women with lifetime PTSD had significantly higher cortisol levels across the day compared to abuse-exposed participants without PTSD, after controlling for age, depression, severity, and latency of abuse. There were no significant group differences in levels of platelet catecholamines. CONCLUSIONS: Elevated cortisol levels may be a biomarker of IPV-related lifetime PTSD, reflecting long lasting changes associated with trauma-exposure or possibly a reflection of risk for PTSD in women. PMID- 16716531 TI - Diurnal salivary cortisol levels in postpartum mothers as a function of infant feeding choice and parity. AB - Daily stress and sleep deprivation can influence the diurnal pattern of cortisol, which normally consists of high morning levels and a gradual decline throughout the day. While most individuals have consistent declining cortisol concentrations over days, others display either flat or inconsistent profiles. Postpartum mothers experience considerable home demands and sleep deprivation, yet, breastfeeding mothers perceive lower stress and reduced negative mood states compared to bottlefeeders. On the other hand, multiparity (having more than one child) is associated with reduced steepness in diurnal cortisol decline. Interestingly, no study to date has investigated the diurnal cortisol pattern and its stability across days in postpartum women as a function of their choice of infant feeding and parity. In this study, we measured salivary cortisol at four different time points during the day, on two non-consecutive days in first-time (primiparous) and second-time (multiparous) mothers at 5-20 weeks postpartum who were exclusively breastfeeding or bottlefeeding, and in non-postpartum mothers of young children (1-6 years). Among multiparous mothers, we found that cortisol levels in those who were bottlefeeding were higher than in breastfeeding mothers at both awakening and 1600 h. This effect remained significant after controlling for individual differences related to infant feeding choice, such as estradiol levels, education and income. No effect of infant feeding choice on cortisol concentrations was observed in primiparous mothers. While a consistent decline across days was common, some mothers presented a flat or inconsistent profile, a profile that was not associated with infant feeding choice or parity. Importantly, mothers with consistent declining profiles had the highest household income. Our findings suggest that although breastfeeding might promote a tighter regulation of diurnal basal cortisol secretion, in particular for multiparous mothers who are likely to be exposed to greater home demands and maternal responsibilities, some aspects of socioeconomic status such as income can also play a significant role in the stability of diurnal cortisol secretion across days. PMID- 16716532 TI - A life cycle for Borrelia spirochetes? AB - Subsequent to Schaudinn and Hoffman's visualization of Treponema pallidum in 1905, many distinguished syphilologists proposed that spirochetes have a life cycle. What is the "essence" of a life cycle? Simply put, life cycles are diverse arrays of life forms, which emerge in an ordered sequence; which are "connected" to one another across primary and secondary hosts, and constitute a cycle with "circular" relationship between hosts. Fecal-oral life cycles and blood-to-blood life cycles are exemplary of host parasite relationships in this realm. The "blood-to-blood" begins and ends with an insect taking a blood "meal". In this operatic scenario, a "blood-less" insect functions simultaneously as a hypodermic needle and as an incubator for some of the infectious components. The initial phase is inside the body fluid compartment of an insect. The second phase is in the blood or body fluid of a warm-blooded mammal. Third, is the phase inside the cell of a mammalian host. And a final portion of the "life" marked by "death" of the parasitized mammalian cells and the release of infectious parasites which return to the "warm" blood where the "cold blooded" vector again takes a blood meal. The cycle then begins again. In each phase of a blood to blood life cycle, the infectious agent changes its shape. Blood phase "profiles" look different from "tissue phase" profiles. Some of the tissue phase profiles may be "invisible". Borrelia spirochetes offer an excellent example of a life cycle, by virtue of the insect vector to mammalian "piece", the blood and intracellular residence "pieces" and the morphologic diversity "piece". Stereotypes of what a spirochete "should " look like, have actually produced a state of "perseveration" in spirochetal pathobiology. We have been "stuck" like a broken record, on the corkscrew form, and have failed to see the rest of the life cycle. Cystic, granular, and cell wall deficient spirochetal profiles, which were well known in the 19th and 20th centuries by such titans as Schaudinn, Hoffman, Noguchi, Delamater, Steiner, and Mattman, have been repudiated by professional microbiologists, and by pathologists who practice and who confer the status of 21st century truths in microbiology matters. Proper microscopic study, as is required by Dr. Robert Koch's second postulate, for establishing links between microbes and disease, presupposes that the microscopist be aware of the complete array of morphologic repertoires of the alleged pathogen. (Morphologies, which are herein introduced.). PMID- 16716533 TI - Botulinum toxin injections to reduce adiposity: possibility, or fat chance? AB - Obese individuals often suffer from negative self-image. Many, even those with a normal body mass index, resort to pharmacotherapy (lipase inhibitors or appetite suppressants), mesotherapy and surgery (gastric volume reduction, liposuction or apronectomy) in a bid to remove excess adipose tissue. These treatments are associated with inherent morbidity and even mortality, and hence should not be undertaken lightly. The observation that denervation of adipose tissue results in lipoatrophy leads us to postulate that chemodenervation using botulinum toxin may achieve the same result, i.e. fat loss, and we explore the methods by which selective fat loss may be achieved. We concede that removal of subcutaneous fat does not, however, reduce the risks associated with the metabolic syndrome, as visceral (intra-abdominal) fat is not reduced by the removal of subcutaneous fat. PMID- 16716534 TI - A reformulation of the hygiene hypothesis: maternal infectious diseases confer protection against asthma in the infant. AB - Epidemiological studies have shown an inverse relationship between allergic respiratory diseases and the number of siblings. It was hypothesized that the lower prevalence of allergic respiratory diseases in large sibships was due to cross-infections between siblings. According to this hygiene hypothesis the increase in the prevalence of atopic diseases is caused by a decrease in the exposure to infections. It was believed that early infections were beneficial for health because of their contribution to the maturation of the immune system. However, in this interpretation a possible protective influence of the mother was overlooked. A new hypothesis is therefore proposed. Maternal exposure to infections induces immunological memory, which protects her children against allergic respiratory diseases. PMID- 16716535 TI - Antimicrobial activity of some endemic plants growing in the Eastern Black Sea Region, Turkey. AB - The Eastern Black Sea Region has an extensive flora because of ample rainfall lasting all year. In this study, antimicrobial effects of 74 crude extracts of 22 endemic plants were investigated. Among the 30 active crude extracts, water insoluble crude extracts from Betula medwediewii, Heracleum platytaenium, Primula longipes, Anthemis cretica ssp. argaea and Centaurea helenioides were the prominent ones with their MIC values. PMID- 16716536 TI - Within-compound associations are not sufficient to produce taste-mediated odor potentiation. AB - Rats were used in two flavor-aversion experiments to determine if within-compound associations could be detected with a taste+odor compound that would not support taste-mediated odor potentation. In Experiment 1, following taste+odor compound conditioning, postconditioning taste extinction significantly weakened the odor aversion. In Experiment 2, following taste+odor compound conditioning, postconditioning taste inflation significantly strengthened the odor aversion. There was no evidence that taste potentiated the odor aversion in either Experiment 1 or 2. Thus, the results demonstrate that the presence of within compound associations is not sufficient to produce taste-mediated odor potentiation. We offer a mediated conditioning explanation to account for the results of these two experiments. PMID- 16716537 TI - Influence of optical cues from conspecific females on escalation decisions during male-male interactions of jumping spiders. AB - An important prediction from game theory is that the value of a resource influences the level to which male-male conflict escalates. An earlier experimental study showed that males of Hypoblemum albovittatum, a common salticid in New Zealand, can discern by sight alone whether a female is in the vicinity and that, having detected a female's presence, males escalate (i.e., adopt behaviour that is likely to put them at greater risk of injury). Seeing a bare moving cork, a conspecific male or a prey item also elicits escalation, but not to the level elicited by seeing a conspecific female. Here the earlier study is extended by using non-conspecific females in control tests and by undertaking experiments with an additional six salticid species. Findings from these experiments suggest that the ability of interacting males to detect by sight alone that a conspecific, instead of a non-conspecific, female is present is widespread in the family Salticidae and that it may be routine for salticid males to use information about the presence of conspecific females when making strategic decisions related to escalation during encounters with other males. PMID- 16716538 TI - A frameshift in the coding region of a novel tomato class I basic chitinase gene makes it a pseudogene with a functional wound-responsive promoter. AB - A putative class I basic chitinase gene, assigned as psiBCH, was cloned from a tomato breeding line NC 24E. The gene contains a coding region with two introns. The predicted psiBCH open reading frame (ORF) is 971 bp and exhibits 81-88% identity at the nucleotide level with known class I basic chitinase genes from the Solanaceae family. However, the presence of a stop codon caused by a frameshift in the ORF of psiBCH makes it unusual among the other class I plant basic chitinases. This stop codon might be involved in the lower accumulation of fully spliced psiBCH RNA caused by nonsense-mediated decay (NMD), which is an RNA surveillance system universally found in eukaryotes. Sequence analysis of the 1883-bp 5'-flanking region of the psiBCH gene revealed the presence of potential wound-response promoter elements. To study the transcriptional regulation of the psiBCH gene, its 5'-flanking region containing the putative promoter was fused to the gus reporter gene and introduced into the tobacco genome via Agrobacterium tumefaciens-mediated transformation. Transgenic plants were functionally assayed for beta-glucuronidase activity. The psiBCH promoter drives the reporter gene expression in response to wounding stimuli. psiBCH promoter-GUS analysis indicates that wound-response of the tobacco transgene was rapid and localized in the wounded area following mechanical wounding. Therefore, our results suggest that the psiBCH promoter can provide targeted expression of genes, such as protease inhibitors in response to pest attack. PMID- 16716539 TI - Origins of sequence diversity in the malaria vaccine candidate merozoite surface protein-2 (MSP-2) in Amazonian isolates of Plasmodium falciparum. AB - The recent evolution of Plasmodium falciparum is at odds with the extensive polymorphism found in most genes coding for antigens. Here, we examined the patterns and putative mechanisms of sequence diversification in the merozoite surface protein-2 (MSP-2), a major malarial repetitive surface antigen. We compared the msp-2 gene sequences from closely related clones derived from sympatric parasite isolates from Brazilian Amazonia and used microsatellite typing to examine, in these same clones, the haplotype background of chromosome 2, where msp-2 is located. We found examples of msp-2 sequence rearrangements putatively created by nonreciprocal recombinational events, such as replication slippage and gene conversion, while maintaining the chromosome haplotype. We conclude that these nonreciprocal recombination events may represent a major source of antigenic diversity in MSP-2 in P. falciparum populations with low rates of classical meiotic recombination. PMID- 16716540 TI - Improved diagnosis of virulent ovine footrot using the intA gene. AB - Footrot is a mixed bacterial infection of the hooves of sheep. The gram-negative anaerobic bacterium Dichelobacter nodosus is the principal causative agent, with different strains causing diseases of different severity, ranging from benign to virulent. In Australia, in the state of New South Wales (NSW), only virulent footrot is subject to regulatory action, including quarantine. However, it is often difficult to distinguish benign footrot from virulent footrot in the initial stages of infection, or under adverse climatic conditions. The gelatin gel test, which measures the thermostability of secreted bacterial proteases, is the laboratory test most widely used in Australia to aid in the differential diagnosis of footrot. The proteases of virulent strains are, in general, more thermostable than the proteases of benign strains. However, there are some false positives in the gelatin gel test, which may lead to unnecessary quarantine procedures. We used Southern blot analysis on 595 isolates of D. nodosus from 124 farms on which sheep had benign or virulent footrot to test for the presence of the intA gene. We found that for D. nodosus strains which are stable in the gelatin gel test, there is a high correlation between the presence of the intA gene and the ability of the strain to cause virulent footrot. We also developed a PCR-based assay for the rapid detection of intA, which can be used to test DNA extracted from colonies grown on plates, or DNA extracted from cotton swabs of culture plates. PMID- 16716542 TI - Letter to the editor: "Is hypotension a major risk factor for neurological morbidity at term age in very preterm infants". PMID- 16716541 TI - The effect of Russian Maternity Home routines on breastfeeding and neonatal weight loss with special reference to swaddling. AB - BACKGROUND: Few investigations have considered evaluating the effects of certain combinations of ward routines like swaddling of the baby and separation of mother and baby on infant variables such as neonatal weight loss. AIMS: To study the effect of different ward routines in respect to proximity to mother and type of infant apparel, on breastfeeding parameters (amount of ingested milk, volume of supplements, number of breastfeeds, total duration of breastfeeding time) day 4 after birth as well as recovery from neonatal weight loss and infant's weight on day 5. STUDY DESIGN AND SUBJECTS: In a randomized trial with factorial design four treatment groups including 176 mother-infant dyads were studied 25-120 min after birth. Randomized treatments focused on care routines administered to the infants after delivery and later in the maternity ward as well as to the type of clothing the infants received. Group 1 infants were placed skin-to-skin with their mothers after delivery, and had rooming-in while in the maternity ward. Group 2 infants were dressed and placed in their mothers' arms after delivery, and roomed-in with mothers in the maternity ward. Group 3 infants were kept in the nursery both after birth and while their mothers were in the maternity ward. Group 4 infants were kept in the nursery after birth, but roomed-in with their mothers in the maternity ward. Equal numbers of infants were either swaddled or clothed in baby attire. Breastfeeding parameters were documented during day 4 after birth. Infant's weight was measured daily. RESULTS: Babies who were kept in the nursery received significantly more formula and significantly less breast milk, than did babies who roomed-in with their mothers. Swaddling did not influence the breastfeeding parameters measured. However, swaddled babies who had experienced a 2-h separation period after birth and then were reunited with their mothers tended to have a delayed recovery of weight loss compared to those infants who were exposed to the same treatment but dressed in clothes. Furthermore, swaddled babies who were kept in the nursery and received breast milk supplements had a significantly delayed recovery of weight loss after birth when compared to those infants ingesting only breast-milk. On day 5, regression analyses of predicted weight gain in the exclusively breastfed infants indicated a significant increase per 100 ml breast-milk (59 g), compared to the predicted weight gain on day 5 per 100 ml supplements in the swaddled babies (14 g) (P=0.001). CONCLUSION: Supplements given to the infants in the nursery had a negative influence on the amount of milk ingested. In addition, supplement feeding or a short separation after birth when combined with swaddling was shown to have a negative consequence to infant weight gain. PMID- 16716543 TI - Comparison of genotoxic and inflammatory effects of particles generated by wood combustion, a road simulator and collected from street and subway. AB - The health effects of exposure to airborne particles are of increasing concern in society. In order to protect public health, a clarification of the toxic properties of particles from different sources is of importance. The aim of this study was to investigate and compare the genotoxicity and the ability to induce inflammatory mediators of nine different particle types from wood and pellets combustion, from tire-road wear and collected from an urban street and a subway station. The comet assay was used to assess genotoxicity after exposure of the human lung cell line A549. Inflammatory effects were measured as induction of IL 6, IL-8 and TNF-alpha after exposure of human macrophages. We found that all particles tested caused DNA damage and those from the subway caused more damage than the other particles (p<0.001) likely due to redox-active iron. In contrast, particles collected from an urban street were most potent to induce inflammatory cytokines. Particles from tire-road wear collected using a road simulator were genotoxic and able to induce cytokines. Finally, more effective combustion of wood led to less emission of particles, but those emitted did not show less toxicity in this study. PMID- 16716544 TI - Culture of mature trophoblastic giant cells from bovine placentomes. AB - The mostly binucleate trophoblast giant cells (TGC) found in bovine placentomes, in addition to synthesizing and releasing hormones play an important role in fetal development and maternal adaptation to pregnancy. Placentomes from early gestation were collected, and for isolation of mature TGC, three cellular disaggregation methods, mechanical (MECH), enzymatic by trypsin (TRYP) or collagenase (COLL) were compared to each other. Further on, the cell survival in culture medium (DMEM) supplemented with either 10% fetal calf serum (FCS) or 10% serum replacement (SR) on culture plates free of any substrate was evaluated over a period of 90 days by trypan blue exclusion. The cells were further characterized by HOECHST 33342 nuclear staining, and immunocytochemical staining with monoclonal antibodies against vimentin and cytokeratin. A mean total rate of TGC survival of 82.56% was recorded. Statistical analysis showed significantly higher survival rates after enzymatic disaggregation with COLL (86.23%) than following MECH (80.38%) or TRYP (80.91%) treatment. Supplementation of DMEM with FCS resulted in significantly higher cellular survival rates (87.13%) when compared to the addition of SR (77.73%). Analysis of the influence of both, disaggregation method and medium supplementation on TGC survival revealed statistically significant differences between the following groups: MECH-SR (71.09%) was significantly lower than all other groups; TRYP-SR (78.03%) was significantly different from all other groups; TRYP-FCS (83.43%) and COLL-SR (84.08%) were significantly lower than MECH-FCS (89.98%) which together with COLL FCS (88.25%) showed the highest cellular survival rate. In summary, our results show that TGC isolated from early gestation placentomes may be viable for more than 90 days of culture. However, whether these TGC produce placental lactogen throughout this period has yet to be determined. PMID- 16716545 TI - The temporal representation of the delay of iterated rippled noise with positive or negative gain by chopper units in the cochlear nucleus. AB - The role of chopper units in representing the pitch of complex sounds is unresolved. Traditionally chopper units have been regarded as primarily responding to the stimulus envelope of complex stimuli. This has been supported by the response of chopper units to iterated rippled noise (IRN) as they can provide a robust representation of the delay of IRN with positive gain (+) in their first-order interspike intervals and for some chopper units this representation is relatively level independent. The envelope modulation of IRN(+), and pitch, is at the reciprocal of the delay, the pitch of IRN with negative gain (IRN(-)) is often at twice the delay. This distinction between IRN(+) and IRN(-) can be used to help determine whether a unit is simply responding to modulation or to stimulus fine structure. Chopper units with relatively high best frequencies (BF) are unable to represent the distinction between IRN(+) and IRN(-). However, in this study it is shown that at least some chopper units, with low BFs (<1.25 kHz), can represent the pitch of the IRN(-) as perceived perceptually. PMID- 16716546 TI - Nephrotoxicity study of total rhubarb anthraquinones on Sprague Dawley rats using DNA microarrays. AB - Total rhubarb anthraquinones (TRAs) are the active therapeutic components from the rhizomes of Rheum palmatum L. (Polygonaceae), which are widely used in traditional Chinese medicines (TCMs) and have been reported to have cell toxicity recently. This study focuses on the toxicity of TRAs on Sprague Dawley (S.D.) rats. TRAs administrated per os for 13 weeks induced nephrotoxicity on S.D. rats as renal tubule epithelial cells swelled and denatured in tissue slice examination. After high-density oligonucleotide microarrays scanning, we have identified mitogen-activated protein kinase (MAPK) kinase 6 to be the target gene which causes cell cycle arrest and proliferation inhibition and contributes to nephrotoxicity on S.D. rats. PMID- 16716547 TI - Lethal paradoxical cerebral vein thrombosis due to suspicious anticoagulant rodenticide intoxication with chlorophacinone. AB - Superwarfarin exposure is a growing health problem, described in many countries. The authors report a case of suspicious chlorophacinone poisoning with a problematic diagnosis. They review the literature and discuss particularities of anticoagulant rodenticide intoxication, as well as the apparent contradiction between anticoagulant intoxication and lethal thrombosis. PMID- 16716548 TI - Calcified chronic subdural hematoma mimicking calvarial mass: a case report. AB - Head trauma is a frequent cause of mortality and morbidity in the pediatric population. Chronic subdural hematoma is the most common traumatic lesion in head injury. Chronic subdural hematomas are rare in children older than 2 years old; they are more frequent during adolescence. Calcified or ossified chronic subdural hematoma is a rare entity that usually presents as a space-occupying lesion over the cerebral convexity. Chronic calcified subdural hematomas are reported less than chronic subdural hematomas. In this article, we report a successfully treated patient with surgical removal case of calcified chronic subdural hematoma mimicking calvarial mass. PMID- 16716549 TI - Amelioration of age-related inflammation and oxidative stress by PPARgamma activator: suppression of NF-kappaB by 2,4-thiazolidinedione. AB - Peroxisome proliferator-activated receptors (PPARs) are members of the nuclear hormone receptor superfamily of transcription factors and are key regulators in various pathophysiological processes related to energy metabolism including lipid and carbohydrate metabolism and inflammation. PPARgamma signaling pathways are reported to exert anti-inflammatory effects by inhibition of NF-kappaB. We previously reported that age-related oxidative stress and inflammatory reactions cause reduced PPARgamma during the aging process. In present study, we investigated the action of 2,4-thiazolidinedione (2,4-TZD), a well-known PPARgamma activator, on aging process using kidneys from Fischer 344 rats, young (9-month-old), old (22-month-old) and old-2,4-TZD fed (4 mg/kg for 10 days). The results showed that the 2,4-TZD treatment brought about several major changes, decrease of: (1) age-related oxidative stress; (2) p65 translocation and NF kappaB binding activity; (3) NF-kappaB-regulated gene expression, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and inflammatory mediators such as interleukin-1beta (IL-1beta), IL-6, adhesion molecules, VCAM-1 and P-selectin; and (4) age-related disturbance of the redox-status. Therefore, we concluded that 2,4-TZD exerted significant anti-oxidative and anti inflammatory effects in aged rats, most likely by its ability to attenuate oxidative stress. We propose that 2,4-TZD or other potent PPARgamma activators may be useful in the therapy against age-related inflammation. PMID- 16716550 TI - The relation between D-galactose injection and mitochondrial DNA 4834 bp deletion mutation. AB - Since,D-galactose (D-gal) overload model has been used as a premature aging model, we hypothesized that it may also lead to accelerated aging in the inner ear. Furthermore, though the mitochondrial DNA (mtDNA) 4834 bp deletion mutation has been considered as the marker of aging, there is no information available in the literature concerning the mtDNA 4834 bp deletion mutation condition of the D gal induced premature aging model. We investigate the changes in inner ear enzymatic activity, the occurring of mtDNA 4834 bp deletion in inner ear and other tissues and the relating hearing thresholds after the administration of high dosage (150 mg/kg per day) and low dosage (50 mg/kg per day) of D-gal to rats. Furthermore, the incidence of the mtDNA 4834 bp deletion in different tissues as well as in blood sample was compared. The results showed that daily subcutaneous injections of D-gal into rats for 8 weeks could lead to the biochemical defects and mtDNA 4834 bp deletion in the inner ear tissue and other tissues, which represent the typical aging animals, but the relating hearing threshold shifts (TS) were nearly identical in the three groups. This study also indicates that using of blood samples to detect mtDNA 4834 bp deletion in clinical research might lead to a 'false negative' result. A higher sensitive result could be gained using tissue biopsy to examine mtDNA 4834 bp deletion. PMID- 16716551 TI - MRI-guided percutaneous cryoablation of renal tumors: use of external manual displacement of adjacent bowel loops. AB - PURPOSE: We sought to investigate retrospectively the safety and effectiveness of using external hand compression to displace adjacent bowel loops during MRI guided percutaneous cryoablation of renal tumors. MATERIALS AND METHODS: Fourteen patients (six women, eight men; mean age: 72 years) with 15 renal tumors (mean diameter: 2.4 cm; range: 1.4-4.6 cm) adjacent to bowel were treated with MRI guided percutaneous cryoablation during which bowel was displaced manually. Bowel loop of concern was ascending colon (n=5), descending colon (n=8), descending colon and small bowel (n=1), ascending colon and small bowel (n=1). To analyze effectiveness of the maneuver, mean distance between tumor margin and bowel before and after the maneuver were compared and analyzed using paired Student's t test. Minimum distance between iceball edge and adjacent bowel with external manual displacement during freezing was also measured. Safety was assessed by analyzing post-procedural MR imaging for adjacent bowel wall thickening and focal fluid collections as well as patients' clinical and imaging follow-up. RESULTS: Mean distance between tumor margin and closest adjacent bowel increased from 0.8 cm (range: 0-2 cm) before external manual compression to 2.6 cm (range: 1.6-4.1 cm) with manual displacement (p<0.01). Mean minimum distance between iceball edge and closest adjacent bowel during the procedures was 1.6 cm (range: 0.5-3.5 cm). No evidence of bowel injury was encountered. Twelve of 15 tumors had follow-up (mean: 10 months) that showed no tumor recurrence. CONCLUSION: MRI-guided percutaneous cryoablation of renal tumors adjacent to bowel can be done safely and effectively using external hand compression to displace bowel loops. PMID- 16716552 TI - Pre-clinical testing of a phased array ultrasound system for MRI-guided noninvasive surgery of the brain--a primate study. AB - MRI-guided and monitored focused ultrasound thermal surgery of brain through intact skull was tested in three rhesus monkeys. The aim of this study was to determine the amount of skull heating in an animal model with a head shape similar to that of a human. The ultrasound beam was generated by a 512 channel phased array system (Exablate 3000, InSightec, Haifa, Israel) that was integrated within a 1.5-T MR-scanner. The skin was pre-cooled by degassed temperature controlled water circulating between the array surface and the skin. Skull surface temperature was measured with invasive thermocouple probes. The results showed that by applying surface cooling the skin and skull surface can be protected, and that the brain surface temperature becomes the limiting factor. The MRI thermometry was shown to be useful in detecting the tissue temperature distribution next to the bone, and it should be used to monitor the brain surface temperature. The acoustic intensity values during the 20 s sonications were adequate for thermal ablation in the human brain provided that surface cooling is used. PMID- 16716553 TI - Magnetic resonance imaging for hepatic radiofrequency ablation. AB - Image-guided radiofrequency (RF) ablation is a minimally invasive therapy option in the treatment of primary and secondary hepatic malignancies. Magnetic resonance (MR) imaging offers an accurate pre-interventional imaging having important impact on patient selection and planning of the ablation procedure. Peri-interventional imaging is used for targeting, monitoring, and controlling of the ablation procedure. Due to a high soft-tissue contrast offering delineation of tumor tissue and the surrounding anatomy, coupled with multiplanar capabilities, MR imaging is an advantageous targeting technique compared with ultrasonography (US) or computed tomography (CT). MR imaging is sensitive to thermal effects enabling a monitoring of ablation therapy subsequently being supportive to control the ablation procedure. Therefore, MR imaging can fulfil the conditions for overlapping ablations by enabling a precise repositioning of the MR compatible RF applicator if required. Thus, the probability of achieving complete coagulation in larger tumors within a single therapy session is potentially increased. A monitoring of thermal effects is moreover essential in order to prevent unintended tissue damage from critical structures in the surrounding of the target tissue. Post-interventional imaging is performed to assess treatment response after RF ablation and has prognostic impact, as an early detection of treatment failure, e.g. residual tumor tissue, enables immediate therapy. Nevertheless, differential diagnostic difficulties arise from benign periablational enhancement which may cover tumor tissue. Hence, further evaluation and improvement in the assessment of treatment response is essential. PMID- 16716554 TI - Optimization of HPLC chromatographic conditions for determination of Transkarbam 12 and its degradation products. AB - This paper deals with searching of HPLC chromatographic conditions for determination and separation of Transkarbam 12 (T 12) and its two main degradation products (omega-aminocaproic acid and dodecylalcohol). T 12 is a new substance which belongs to the group of accelerators of transdermal penetration. Chromatographic separation was achieved using Separon SGX C18 analytical column (150 mm x 3 mm i.d.; 5 microm). Mobile phase contained acetonitrile and sodium acetate buffer pH 4.5 at the flow rate of 1 ml/min. Separation was carried out under the conditions of gradient elution. After the modification of the structure by derivatization reagent (3,5-dinitrobenzoyl chloride) detection at wavelength 230 nm was realized. The aim of this study was not only the optimization of the separation of derivatization reagent and derivatized T 12, Ak and D but also optimal derivatization processes for all three substances. PMID- 16716555 TI - Determination of paracetamol: historical evolution. AB - Paracetamol is a common analgesic and antipyretic drug that is used for the relief of fever, headaches and other minor aches and pains. Their determination in pharmaceuticals is of paramount importance, since an overdose of paracetamol can cause fulminating hepatic necrosis and other toxic effects. Many analytical methodologies have been proposed for the determination of paracetamol. The aim of the present study is to evaluate the utility of different techniques for quantification of paracetamol content in pharmaceutical formulations and biological samples. PMID- 16716556 TI - Industrial and biotechnological applications of laccases: a review. AB - Laccases have received much attention from researchers in last decades due to their ability to oxidise both phenolic and non-phenolic lignin related compounds as well as highly recalcitrant environmental pollutants, which makes them very useful for their application to several biotechnological processes. Such applications include the detoxification of industrial effluents, mostly from the paper and pulp, textile and petrochemical industries, use as a tool for medical diagnostics and as a bioremediation agent to clean up herbicides, pesticides and certain explosives in soil. Laccases are also used as cleaning agents for certain water purification systems, as catalysts for the manufacture of anti-cancer drugs and even as ingredients in cosmetics. In addition, their capacity to remove xenobiotic substances and produce polymeric products makes them a useful tool for bioremediation purposes. This paper reviews the applications of laccases within different industrial fields as well as their potential extension to the nanobiotechnology area. PMID- 16716557 TI - Carica papaya lipase (CPL): an emerging and versatile biocatalyst. AB - In recent years, the Carica papaya lipase (CPL) is attracting more and more interest. This hydrolase, being tightly bonded to the water-insoluble fraction of crude papain, is thus considered as a "naturally immobilized" biocatalyst. To date, several CPL applications have already been described: (i) fats and oils modification, derived from the sn-3 selectivity of CPL as well as from its preference for short-chain fatty acids; (ii) esterification and inter esterification reactions in organic media, accepting a wide range of acids and alcohols as substrates; (iii) more recently, the asymmetric resolution of different non-steroidal anti-inflammatory drugs (NSAIDs), 2 (chlorophenoxy)propionic acids, and non-natural amino acids. Taking into account the novelty and the current interest of the topic, this review aims to highlight the origin, features, and applications of the C. papaya lipase, with the objective to prompt research groups to further investigate the spectra of applications that this emerging and versatile CPL could have in the future. PMID- 16716558 TI - [Hypothermia after spinal anaesthesia: errors in temperature measurement]. PMID- 16716559 TI - [Eating during labour: is it of any benefit?]. AB - Fasting during labour is questioned in France despite the historical recommendations by Curtis Mendelson. Solid food diet increases maternal nausea and vomiting of non digested food associated with a theoretical risk of severe aspiration syndrome. Clear fluids may improve the comfort of some parturients but it remains uncertain whether or not the obstetric consequences (i.e. duration of labour, Caesarean section rate) of carbohydrate supplementation are beneficial. PMID- 16716560 TI - [Obstructive pyocholecystitis and septicaemia related from Hafnia alvei: influence of diabetes mellitus]. PMID- 16716561 TI - Mitogen activated protein kinase p38 pathway is an important component of the anti-inflammatory response in Mycobacterium avium subsp. paratuberculosis infected bovine monocytes. AB - We investigated the role of cell signaling through the mitogen-activated protein kinase-p38 (MAPK p38) pathway on the antimicrobial functions and cytokine expression by bovine monocytes after ingestion of Mycobacterium avium subsp. paratuberculosis. We evaluated the dynamic secretion of interleukin (IL)-10, IL 12 and tumor necrosis factor-alpha (TNF-alpha) as well as phagosome acidification and organism killing at several time points after in vitro infection of bovine monocytes with M. avium subsp. paratuberculosis. Monocytes treated with M. avium subsp. paratuberculosis had a significant increase in IL-10 expression at 2, 4, and 6h post-infection and an increase expression of TNF-alpha at 2, 4, 6, and 24h post-infection. In contrast, IL-12 expression did not increase at any time point post-infection. Moreover, MAPK p38 was rapidly phosphorylated at 10 and 60 min after M. avium subsp. paratuberculosis ingestion. Chemical inhibition of the MAPK p38 signaling pathway (SB203580) resulted in decreased expression of IL-10 and increased expression of IL-12 at 6h post-infection. Chemically blocking the MAPK p38 pathway also increased acidification of phagosomes as well as increasing the capacity of macrophages to kill organisms. Taken together, these results indicated that selective activation of MAPK p38 may be a major mechanism exploited by M. avium subsp. paratuberculosis to circumvent the antimycobacterial effects of mononuclear phagocytes. PMID- 16716562 TI - Evaluation of antimutagenic activity and mechanisms of action of beta-glucan from barley, in CHO-k1 and HTC cell lines using the micronucleus test. AB - Due to the need to identify new antimutagenic agents and to determine their mechanism of action, the present study examined the mechanism of action of the beta-glucan with regard to antimutagenicity using the micronucleus assay in CHO k1 and HTC cell lines. The mutagenicity experiments were performed with three different concentrations of beta-glucan (5, 10, and 20 microg/mL), in wich only the highest dose showed mutagenic activity. In the antimutagenicity experiments, the same concentrations of beta-glucan were combined with a mutagenic agent, methylmethane sulfonate, or 2-aminoanthracene, using four different treatment protocols: pre-treatment, simultaneous treatment (simple and with pre incubation), and post-treatment. The results indicate that the CHO-k1 cell line treated with MMS presented a chemopreventive activity for all the doses of beta glucan in the different treatment protocols, except for the lowest dose in post treatment. When HTC cell line treated with MMS is analysed, a chemopreventive activity can be verified for the highest dose in both pre- and post-treatment. For the simple simultaneous treatment, the three doses demonstrated efficacy, while for the simultaneous treatment with pre-incubation only the intermediate concentration was effective. In HTC treated with 2AA both the lowest dose in the pre-treatment protocol and the post-treatment protocol did not show efficacy in preventing DNA damage. The evaluation of the different protocols and the damage decrease percentages observed suggest that beta-glucan has both desmutagenic and bioantimutagenic activity. It is necessary, however, to note that efficacy and mechanism of action are subject to variation when compared the two cell lines, since in HTC, representing a drug-metabolizing system, this substance can show a diminished chemopreventive capacity. PMID- 16716563 TI - Cognitive functioning and postconcussive symptoms in trauma patients with and without mild TBI. AB - Although there is a large body of research on mild traumatic brain injury (MTBI), the portion that pertains to acute patients (those less than 1 month postinjury) is relatively small and yields inconsistent findings. The potential contribution of non-neurological factors, such as pain and emotional distress, to the clinical picture in this population is also lacking. To address these issues, the cognitive performance and symptom complaints of 37 hospitalized MTBI subjects were compared to those of 39 hospitalized trauma subjects, averaging 4.5 days postinjury. MTBI subjects performed significantly worse on all cognitive measures, but did not differ from trauma subjects in their report of postconcussive symptoms. Analyses also revealed that cognitive performance was unrelated to pain severity and emotional distress. Postconcussive symptoms were similarly unrelated to pain severity, but were consistently related to emotional distress. Results are discussed in terms of their etiological and treatment implications. PMID- 16716564 TI - Model-based identification of cis-acting elements from microarray data. AB - Identification of transcriptional regulatory motifs continues to be a challenging problem in computational biology. We report a model-based procedure, MotifModeler, that uses global gene expression data to (1) identify cis-acting elements (CAE) that regulate gene expression under a given condition and (2) estimate the effects of the CAE on gene expression. MotifModeler repeatedly tests random subsets of all possible motifs of a given size and selects those that best fit a combinatorial model of the expression levels. We tested MotifModeler using data from a microarray experiment on the effects of interferon-alpha in peripheral blood monocytes. Focusing on 6-bp motifs, we predicted 16 stimulatory and 4 inhibitory motifs. Motifs were extended and compared to known binding sites in the TRANSFAC database using position-specific scoring matrices. Many predicted CAE match sites known to be involved in interferon action. MotifModeler demonstrated the potential to computationally identify CAE important in gene regulation. PMID- 16716565 TI - Reproductive and developmental toxicity of inhaled 2,3-dichloro-1,3-butadiene in rats. AB - Inhalation developmental and reproductive toxicity studies were conducted with 2,3-dichloro-1,3-butadiene (DCBD), a monomer used in the production of synthetic rubber. In the reproductive toxicity study, Crl:CD(SD)IGS BR rats (24/sex/group) were exposed whole body by inhalation to 0, 1, 5, or 50 ppm DCBD (6 h/day) for approximately 10-11 weeks total, through premating (8 weeks; 5 days/week), cohabitation of mating pairs (up to 2 weeks, 7 days/week), post-cohabitation for males (approximately 7 days) and from conception to implantation (gestation days 0-7 [GD 0-7]), followed by a recovery period (GD 8-21) for presumed pregnant females. Estrous cyclicity was evaluated during premating (last 3 weeks) and cohabitation. Reproductive organs and potential target organs, sperm parameters, and GD 21 fetuses (viability, weight, external alterations) were evaluated. In the developmental study, pregnant Crl:CD(SD)IGS BR rats (22/group) were exposed whole body by inhalation to 0, 1, 10, or 50 ppm DCBD (6 h/day) on GD 6-20; dams were necropsied on GD 21 (gross post-mortem only) and fetuses were evaluated (viability, weight, and external, visceral and skeletal exams). During the in life portion of the studies, body weight, food consumption, and clinical observation data were collected. At 50 ppm, gasping and labored breathing occurred in both studies during the first few exposures; body weight and food consumption parameters were affected in parental animals from both studies, but were more severely affected in the developmental study. Fetal weight was decreased in the developmental study at 50 ppm. Degeneration of the nasal olfactory epithelium was observed in the reproduction study at 50 ppm. There were no effects on reproductive function, embryo-fetal viability, or increases in fetal structural alterations in either study. The no-observed-adverse-effect level (NOAEL) for reproductive toxicity was 50 ppm. The NOAEL for systemic toxicity in the reproduction study was 5 ppm based on adverse effects on body weight and food consumption parameters and nasal olfactory epithelial toxicity at 50 ppm in parental rats. The NOAEL for maternal and developmental toxicity was 10 ppm based on reduced maternal weight gain and food consumption and reduced fetal weight at 50 ppm in the developmental toxicity study. PMID- 16716566 TI - AFLP fingerprinting for the analysis of genetic diversity within Necator americanus. AB - In the present study, we utilised the method of AFLP to screen for genetic variation within and among individuals of the blood-feeding human hookworm Necator americanus (Nematoda) from Africa, Asia and South America. A total of 45 adult worms (i.e. 20 from Ghana, 16 from Colombia and 9 from Nepal) were subjected to analysis using the restriction enzyme/primer combination HindIII+AG/BglII+AC. Cluster analysis divided N. americanus into multiple, genetically distinct groups, consistent with previous findings using ribosomal and mitochondrial DNA data sets. The results demonstrated the usefulness of AFLP fingerprinting for establishing genetic variation within N. americanus and reinforce its applicability to other parasitic helminths of human and/or veterinary health importance. PMID- 16716567 TI - G protein-coupled receptors for free fatty acids. AB - Free fatty acids (FFAs) are not only an important direct source of energy but they also play key roles in regulating a range of physiological responses. Although it was long assumed that such effects of FFAs must occur following cellular uptake, and potentially via their conversion to fatty acyl-CoAs, it is now apparent that FFAs also function directly as agonists at a number of G protein-coupled receptors (GPCRs). Tissue distribution studies and, subsequently, siRNA knock-down experiments have indicated key roles for these GPCRs in glucose homeostasis, adipogenesis, white cell recruitment and potentially in a range of other processes. Considerable interest is thus now centred on the generation of potent and selective small molecule ligands, both as tool compounds to further unravel the biology and physiological role of this group of GPCRs and as starting points for possible therapeutic intervention in a range of areas, particularly those associated with 'metabolic syndrome'. PMID- 16716568 TI - Are deficits in executive sub-processes simply reflecting more general cognitive decline in schizophrenia? AB - BACKGROUND: Schizophrenia is associated with both global and specific cognitive deficits. We sought to investigate whether deficits in executive subcomponents differed in their relationship to global cognitive impairments. METHOD: 95 patients were classified according to pre-morbid and current general cognitive ability as having either (a) intact pre-morbid and current general cognitive ability; (b) intact pre-morbid but deteriorated current ability, and (c) deteriorated both pre-morbid and current cognitive ability. All patients completed measures of verbal and spatial working memory, sustained selective attention, attentional set sifting, and inhibitory control. RESULTS: Deficits on both measures of working memory were associated with general cognitive ability. None of the attentional control deficits observed were associated with general ability. Further, spatial working memory deficits were also associated with more severe negative symptoms. CONCLUSIONS: These results provide further evidence of the discreet nature of attentional deficits in schizophrenia. By contrast, this study suggests that working memory deficits may to some extent index more general cognitive decline. Awareness of such overlap is important for schizophrenia genetics studies where working memory measures has been used to index supposedly discreet aspects of cognitive dysfunction. PMID- 16716569 TI - Ligation of E-cadherin molecules on murine resident Langerhans cells inhibits their maturation and chemokine production. PMID- 16716570 TI - New plasmid-mediated AmpC beta-lactamase (CMY-21) in Escherichia coli isolated in the UK. PMID- 16716571 TI - Quinine distribution in pregnant mice with Plasmodium berghei malaria. AB - Maternal malaria is associated with placental insufficiency that leads to intrauterine growth retardation and reduced birth weight. Malaria may impair the exchange of drugs across the placenta especially the transmission of antimalarial drugs to the foetus. The distribution of quinine and its 3-hydroxymetabolite in blood, tissues and foeto-placental unit was evaluated on day 18 of pregnancy of mice infected or not with Plasmodium berghei. During pregnancy, quinine distribution volume increases gradually with the rise of free quinine concentrations in plasma. Quinine concentrations increase in erythrocytes and most tissues without change in systemic clearance. A maternal-to-foetal gradient of 8:1 limits the exposure of foetus to quinine. During malaria, the systemic clearance of quinine and the 3-hydroxyquinine gradually decrease with the rising parasitaemia. Quinine concentrations increase slightly in most of the tissues. The weight of placentas decreases in a parasitaemia-dependant manner and is strongly related to the low uptake of quinine by placenta. Foetal weights and quinine concentrations in foetus only decrease for the highest parasitaemia. In this experimental model, pregnancy facilitates quinine uptake by erythrocytes and peripheral tissues. Malaria induces a hypotrophy of both placenta and foetus. In placenta, the marked decrease of quinine concentrations may impair the clearance of sequestered parasites. PMID- 16716572 TI - Pharmacological properties of the ubiquitous natural product betulin. AB - Betulin (lup-20(29)-ene-3beta,28-diol) is an abundant naturally occurring triterpene and it is found predominantly in bushes and trees forming the principal extractive (up to 30% of dry weight) of the bark of birch trees. Presently, there is no significant use for this easily isolable compound, which makes it a potentially important raw material for polymers and a precursor of biologically active compounds. Betulin can be easily converted to betulinic acid, which possesses a wide spectrum of biological and pharmacological activities. Betulinic acid has antimalarial and anti-inflammatory activities. Betulinic acid and its derivatives have especially shown anti-HIV activity and cytotoxicity against a variety of tumor cell lines comparable to some clinically used drugs. A new mechanism of action has been confirmed for some of the most promising anti HIV derivatives, which makes them potentially useful additives to the current anti-HIV therapy. Betulinic acid is specifically cytotoxic to several tumor cell lines by inducing apoptosis in cells. Moreover, it is non-toxic up to 500 mg/kg body weight in mice. The literature concerning derivatization of betulin for structure-activity relationship (SAR) studies and its pharmacological properties is reviewed. PMID- 16716573 TI - Motor imagery. AB - We describe general concepts about motor imagery and differences to motor execution. The problem of controlling what the subject actually does during imagery is emphasized. A major part of the chapter is dealing with mental training by imagery and the usage of motor imagination in athletes, musicians and during rehabilitation. Data of altered representations of the body after loss of afferent information and motor representation due to limb amputation or complete spinal cord injury are demonstrated and discussed. Finally we provide an outlook on additional work about motor imagery important for further understanding of the topic. PMID- 16716574 TI - [Epidemiological features and complications of central venous catheters in pediatric oncology: prospective study about 125 cases]. AB - OBJECTIVE: To report the indications and early and late catheter-related complications in a Tunisian unit of paediatric oncology. PATIENTS AND METHODS: This prospective study has been performed in a paediatric oncology unit of the Salah Azaiz Institute between 1989 and 2005. It concerns 58 girls and 57 boys with a median age of 7.9 years(4 months to 18 years) treated for cancer disease predominantly lymphoma (22%), sarcoma (23.2%) or leukaemia (8.5%) proposed for insertion of a central venous catheter (CVC). RESULTS: Excluding 2 insertion failures (1.6%), we placed 123 CVC (double for 10 patients), 43 (35%) exteriorised (EC) and 80 (65%) connected to an implantable site (IS). Catheters were placed in the subclavian vein in 59.2% of cases vs 32% for internal jugular vein and 8.8% for femoral vein. Early complications included 15 cases of multiple punctures (12%), 4 cases of pneumothorax (3.2%) and 6 of arterial punctures (4.8%) originating a cervical subcutaneous haematoma in 1 patient (0.8%). Late complications were represented by infection in 7 cases (5.7%). We observed 2 cases of intracardiac catheter migration due to catheter disconnection from the IS (1.6%) and 2 cases of thrombosis (1.8%). The mean life of CVC was longer for IS (305.2 days) than for EC (64.4 days). CONCLUSION: Implantable sites are effective progress for venous access in children with cancer. They improved the quality of care in pediatric oncology. PMID- 16716575 TI - [Dysfunctional parenting: prevention and care]. AB - This article exposes the work realised by a child psychiatry organization of prevention cares in pre- or post-partum, based on an intricately articulated network of professionals. The author mentions the signals likely to induce the establishment of a well-coordinated network whose effects will make care easier and allow for the creation of new links between professionals/parents; parents/child. We founded our observations on the practical case of a mother granted complete follow-up and attention from the beginning of pregnancy. The quality of exchanges demonstrates how these links between professionals can create a positive step into evolution of parenthood. PMID- 16716576 TI - [Necrotizing pneumonia in children: apropos of 4 cases]. AB - OBJECTIVES: To describe the necrotizing pneumonia in children, a severe affection which prevalence seems to increase; to review literature. PATIENTS AND METHODS: We report 4 cases of necrotizing pneumonia: symptoms, agents, diagnostic tools, treatment and long term evolution. RESULTS: In 2 cases, pneumatoceles could be seen at chest X-ray. Two patients presented a deficiency of anti-pneumococcal antibodies. Three needed insertion of a pleural chest tube of whom 1 had a resection of a small piece of necrotic lung. Duration of hospitalisation is longer than in uncomplicated pneumonias. CONCLUSION: Necrotizing pneumonia is a severe affection. Diagnosis has to be made by lung CT. Long term evolution is excellent in pediatric population with serious management at hospital. PMID- 16716577 TI - [Neurofibromatosis type 1 complications in the pediatric age: follow-up of a hundred cases]. AB - Neurofibromatosis 1 (NF1) is a frequent genetic disease. Diagnostic criterias were established in 1988. The patients can exhibit various and unpredictable complications. OBJECTIVES: To check the efficiency of a coordinated follow-up in specialized multidisciplinary centers providing a higher quality of management and to have a better knowledge of the complications including their true frequencies. POPULATION AND METHODS: We report a serie of 100 NF1 children who were followed-up during 4 years in a specialized center at the Tours University Hospital. Three hospital check-up at 2-5, 6-7, 14-15 years of age were performed as well as an annual physical examination. RESULTS: In our serie, the mean age was 7.8 years old with a sex ratio of 1. The mean age at diagnosis was 3.8 years old and the main diagnosis criteria were the cafe-au-lait spots and the family history for 80% of the patients. The optic nerve glioma has a low frequency of 5%. Learning disabilities clearly represent the most frequent complication (46% of the patients). CONCLUSION: An early detection of these difficulties is a priority for the appropriate management of these children. PMID- 16716578 TI - [Gastroesophageal reflux in childhood]. AB - Gastro-oesophageal reflux (GOR) is a physiological event defined as flow of gastric content into oesophagus. GOR is frequent in infants commonly subjected to regurgitation. While this symptom usually disappears within the first year of life, there is evidence that some symptoms may persist during childhood. In non complicated GOR, no investigation is indicated. PH-metry is only indicated in case of doubtful diagnosis or extradigestive manifestation. Upper gastro intestinal endoscopy is indicated when oesophagitis is suspected. First intention treatment is no intervention or dietetic measures. Thickened feeding reduces symptoms such as regurgitation but does not influence GOR. Except for cisapride which use is nowadays very limited, no prokinetics have demonstrated their efficacy on GOR. Anti-secretory drugs should be restricted to the treatment of oesophagitis. Investigations are necessary before prolonged use. PMID- 16716579 TI - Mammographic masses characterization based on localized texture and dataset fractal analysis using linear, neural and support vector machine classifiers. AB - OBJECTIVE: Localized texture analysis of breast tissue on mammograms is an issue of major importance in mass characterization. However, in contrast to other mammographic diagnostic approaches, it has not been investigated in depth, due to its inherent difficulty and fuzziness. This work aims to the establishment of a quantitative approach of mammographic masses texture classification, based on advanced classifier architectures and supported by fractal analysis of the dataset of the extracted textural features. Additionally, a comparison of the information content of the proposed feature set with that of the qualitative characteristics used in clinical practice by expert radiologists is presented. METHODS AND MATERIAL: An extensive set of textural feature functions was applied to a set of 130 digitized mammograms, in multiple configurations and scales, constructing compact datasets of textural "signatures" for benign and malignant cases of tumors. These quantitative textural datasets were subsequently studied against a set of a thorough and compact list of qualitative texture descriptions of breast mass tissue, normally considered under a typical clinical assessment, in order to investigate the discriminating value and the statistical correlation between the two sets. Fractal analysis was employed to compare the information content and dimensionality of the textural features datasets with the qualitative information provided through medical diagnosis. A wide range of linear and non linear classification architectures was employed, including linear discriminant analysis (LDA), least-squares minimum distance (LSMD), K-nearest-neighbors (K nn), radial basis function (RBF) and multi-layer perceptron (MLP) artificial neural network (ANN), as well as support vector machine (SVM) classifiers. The classification process was used as the means to evaluate the inherent quality and informational content of each of the datasets, as well as the objective performance of each of the classifiers themselves in real classification of mammographic breast tumors against verified diagnosis. RESULTS: Textural features extracted at larger scales and sampling box sizes proved to be more content-rich than their equivalents at smaller scales and sizes. Fractal analysis on the dimensionality of the textural datasets verified that reduced subsets of optimal feature combinations can describe the original feature space adequately for classification purposes and at least the same detail and quality as the list of qualitative texture descriptions provided by a human expert. Non-linear classifiers, especially SVMs, have been proven superior to any linear equivalent. Breast mass classification of mammograms, based only on textural features, achieved an optimal score of 83.9%, through SVM classifiers. PMID- 16716580 TI - Omega 6 to omega 3 fatty acid imbalance early in life leads to persistent reductions in DHA levels in glycerophospholipids in rat hypothalamus even after long-term omega 3 fatty acid repletion. AB - Failure to provide omega 3 fatty acids in the perinatal period results in alterations in nerve growth factor levels, dopamine production and permanent elevations in blood pressure. The present study investigated whether changes in brain (i.e., hypothalamus) glycerophospholipid fatty acid profiles induced by a diet rich in omega 6 fatty acids and very low in alpha-linolenic acid (ALA) during pregnancy and the perinatal period could be reversed by subsequent feeding of a diet containing ALA. Female rats (6 per group) were mated and fed either a low ALA diet or a control diet containing ALA throughout pregnancy and until weaning of the pups at 3 weeks. At weaning, the pups (20 per group) remained on the diet of their mothers until 9 weeks, when half the pups were switched onto the other diet, thus generating four groups of animals. At 33 weeks, pups were killed, the hypothalamus dissected from the male rats and analysed for glycerophospholipid fatty acids. In the animals fed the diet with very little ALA and then re-fed the control diet containing high levels of ALA for 24 weeks, the DHA levels were still significantly less than the control values in PE, PS and PI fractions, by 9%, 18% and 34%, respectively. In this group, but not in the other dietary groups, ALA was detected in all glycerophospholipid classes at 0.2-1.7% of the total fatty acids. The results suggest that omega 6-3 PUFA imbalance early in life leads to irreversible changes in hypothalamic composition. The increased ALA and reduced DHA proportions in the animals re-fed ALA in later life are consistent with a dysfunction or down-regulation of the conversion of ALA to 18:4n-3 by the delta-6 desaturase. PMID- 16716583 TI - A choice experiment of the residential preference of waste management services - the example of Kagoshima city, Japan. AB - Municipal governments have attempted to reduce waste disposal and increase recycling rates in Japan. However, it is difficult to get full cooperation from residents. Using experiments in which residents are given an opportunity to choose the components of the waste management system (choice experiments), we measure the cost of each characteristic of the waste collection services. The estimation result reveals that there are trade-offs between the risk, payments, and handling costs. The marginal loss in utility revenues from an increase in the types of waste being separated is almost 200 yen (US$ 1.74) and a 1% increase in the recycling rate raises the cost of service by 53 yen (US$ 0.46). In addition, the results show the need for communications about risks between municipal authorities and residents. PMID- 16716584 TI - Catalytic detoxification of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans in fly ash. AB - An efficient catalytic detoxification method for polychlorinated dibenzo-p dioxins and polychlorinated dibenzofurans (PCDD/Fs) in fly ash produced by municipal waste incinerator has been studied using palladium on carbon (Pd/C) catalyst. As one of the trace components in fly ash, the detoxification of PCDD/Fs is very difficult because of the interferences of other persistent components with higher concentrations. However, the detoxification reaction of PCDD/Fs shows higher activity in water/isopropanol solution using commercial Pd/C catalyst at 40 degrees C under normal pressure. The results indicated that the catalytic degradation of PCDFs has been found to be easier than that of PCDDs. Moreover, the dechlorination ratios were higher for octa- and hepta-chlorinated congeners than those for tetra- and penta-chlorinated ones. The detoxification process worked well in water. The dechlorination efficiencies of almost all of the PCDD/Fs congeners can reach over 99% within a shorter reaction time. PMID- 16716585 TI - Nanogold hollow balls with dendritic surface for hybridization of DNA. AB - Au hollow balls are fabricated by adsorption of gold 3.5 nm in diameter onto a mixed vesicle composed of mixed polymerized diacetylene which made of negative charged 10,12-pentacosadiynoic acid (PCDA) and positive charged 10,12 pentacosadiynoic acid 2'-aminoethylamide (PCDA-NH(2)). The morphology of these hollow spheres could be controlled by changing the ratio of PCDA and PCDA-NH(2) and the immobilization and hybridization ability of the gold hollow ball have been investigated using a quartz crystal microbalance (QCM). It was found that a dendritic surface in an appropriate ratio existed. The hybridization amount of target DNA is about three to five times for the Au-mixed hollow ball at an optimal ratio (PCDA/PCDA-NH(2)=1/3) as compared with that for pure Au-PCDA-NH(2), though the immobilization amount of ssDNA on these two samples are almost the same, and the detected limitation of target DNA is extend from 10(-9) to 10(-12) M. The stability against breakage by transportation, combined with the simplicity and efficiency of detection, would offer an important advantage over unpolymerized one. This result shows the possibility to control the morphology and surface of nanogold hollow spheres by changing the ratio of PCDA and PCDA NH(2) for the develop of a better DNA detection assay, further proving the idea that low surface coverage and higher DNA probe to target DNA ratios lead to optimal hybridization. PMID- 16716586 TI - microAg particle-based molecular sensing/recognition via surface-enhanced Raman spectroscopy. AB - In this study, we demonstrate that powders of commercially available 2-microm sized Ag (microAg) can be used as a core material for constructing molecular sensing/recognition units operating via surface-enhanced Raman scattering (SERS). This is possible because microAg powders are very efficient substrates for both the infrared and Raman-spectroscopic characterization of molecular adsorbates prepared in a similar manner on silver surfaces. The Raman spectrum of organic monolayers on powdered silver is a SERS spectrum. The agglomeration of microAg particles in a highly concentrated buffer solution could be prevented by the deposition of polar molecules like 1,4-phenylenediisocyanide (1,4-PDI), and mixed self-assembled monolayers of 1,4-PDI and N-(+)-biotinyl-6-aminocaproic acid on microAg particles were then confirmed via the SERS of 1,4-PDI to selectively recognize the avidin arrays formed on a separate biotinylated substrate. According to a dose response curve, avidin at >10(-6)g/mL could be easily identified by the present method. In addition, the non-specific adsorption of microAg particles was found to be negligibly small, probably because the Ag particles were too heavy to be retained on organic substrates solely by non specific interaction. PMID- 16716587 TI - Physico-chemical method for ammonia removal from synthetic wastewater using limestone and GAC in batch and column studies. AB - The purpose of the present study was to examine the removal of ammoniacal nitrogen (NH4-N) from synthetic wastewater using limestone (LS) and granular activated carbon (GAC) mixture as low cost adsorbent. In batch study, optimum shaking and settling times were 150 and 120 min, respectively. The LS-GAC mixture ratio of 25:15 removed about 58% NH4-N. The smaller particle size of medium yielded higher adsorption capacity. The equilibrium adsorption data followed the Freundlich isotherm (R2 > 0.98) but it showed weak bond. Adsorption kinetics were well described by the pseudo second-order rate model (R2 > 0.93). The upflow column showed that higher flow rate and initial concentration resulted in shorter column saturation time. The study showed that the usage of GAC could be reduced by combining GAC with LS for the removal of NH4-N from wastewater; thus reducing the cost of treatment. PMID- 16716588 TI - A gatekeeper residue for inhibitor sensitization of protein tyrosine phosphatases. AB - Allele-specific enzyme inhibitors are powerful tools in chemical biology. However, few general approaches for the discovery of such inhibitors have been described. Herein is reported a method for the sensitization of protein tyrosine phosphatases (PTPs) to small-molecule inhibition. It is shown that mutation of an active-site isoleucine to alanine (I219A) sensitizes PTP1B to inhibition by a class of thiophene-based inhibitors. This sensitization strategy succeeds for both 'orthogonal' inhibitors, designed to be incompatible with wild-type PTP active sites, and previously optimized wild-type PTP inhibitors. The finding that the I219A mutation sensitizes phosphatase domains to a variety of compounds suggests that isoleucine 219 may act as a 'gatekeeper' residue that can be widely exploited for the chemical-genetic analysis of PTP function. PMID- 16716589 TI - Effect of substitution on novel tricyclic HIV-1 integrase inhibitors. AB - A series of novel tricyclic inhibitors of HIV-1 integrase enzyme was prepared. The effect of substitution at C-6 of the 9-hydroxy-6,7-dihydropyrrolo[3,4 g]quinolin-8-one compounds was studied in vitro. Inhibitors with small side chains at C-6 were generally well tolerated by the enzyme, and the physicochemical properties of the inhibitors were improved by substitution of a small alkyl group at this position. A second series of analogs bearing a sulfamate at the C-5 position with various C-6 substituents were prepared to explore the interplay between the two groups. The SAR of the two classes are not parallel; modification at C-5 impacts the effect of substitutions at C-6. PMID- 16716590 TI - A data model to predict HER2 status in breast cancer based on the clinical and pathologic profiles of a large patient population at a single institution. AB - Recently published clinical trial data have produced compelling evidence for increased survival when Herceptin is administered to patients whose tumors are HER2 amplified. Therefore, the accuracy of HER2 status is essential to determine which patients should or should not receive Herceptin. Although HER2 results obtained by FISH and IHC are often in agreement, there is a persistent group of cases in which results are discordant, particularly among tumors with intermediate results. A multivariable analysis was undertaken to determine relative significance of various clinical and pathologic findings for patients diagnosed with infiltrating ductal carcinoma, and a data model was produced that predicts which patients are most likely to have HER2 amplified tumors. Correlates of HER2 amplification were higher Scarff-Bloom-Richardson grade, younger age at diagnosis, and a comedo ductal carcinoma in situ component. PMID- 16716591 TI - Endosomal and non-endosomal functions of ESCRT proteins. AB - The three endosomal sorting complexes required for transport (ESCRTs) are integral to the degradation of endocytosed membrane proteins and multivesicular body (MVB) biogenesis. Here, we review evidence that ESCRTs have evolved as a specialized machinery for the degradative sorting of ubiquitinated membrane proteins and we highlight recent studies that have shed light on the mechanisms by which these complexes mediate protein sorting, MVB biogenesis, tumour suppression and viral budding. We also discuss evidence that some ESCRT subunits have evolved additional functions that are unrelated to membrane trafficking. PMID- 16716592 TI - Dynamometric analysis of the maximum force applied in aquatic human gait at 1.3m of immersion. AB - BACKGROUND: This work had the objective to analyze the values of the vertical and anteroposterior components of the ground reaction force (GRF) during the aquatic gait and the influence of the speed and the upper limb position on the GRF components values. METHODS: Sixty subjects, with average height between 1.6 and 1.85m and average age of 23 years, were divided in three groups according to the immersion level. The citizens walked over a walking platform, which had two force plates attached. The platform was located at a depth of 1.3m. The subjects walked over the platform in four different situations, with speed and upper limb position variations. For data analysis, descriptive and inferential statistics were used. FINDINGS: For the vertical component, the force values varied between 20% and 40% of the subjects' body weight according to the different data collection situations. For the anteroposterior component, the force values reached between 8% and 20% of the subjects' body weight corporal, also according with the data collection situation. INTERPRETATION (DISCUSSION): It was noted that for a given immersion level, the forces can vary according to the request that is imposed to the aquatic gait. It was concluded that either the speed as well as the position of the upper limb influence the values of the GRF components. An increase in the gait speed causes increase of the anteroposterior component (Fx), while an increase in the corporal mass out of the water causes increase mainly of the vertical component (Fy). Knowing the value of these alterations is important for the professional who prescribes activities in aquatic environment. PMID- 16716593 TI - Molecular chaperones: assisting assembly in addition to folding. AB - The common perception that molecular chaperones are involved primarily with assisting the folding of newly synthesized and stress-denatured polypeptide chains ignores the fact that this term was invented to describe the function of a protein that assists the assembly of folded subunits into oligomeric structures and only later was extended to embrace protein folding. Recent work has clarified the role of nuclear chaperones in the assembly of nucleosomes and has identified a cytosolic chaperone required for mammalian proteasome assembly, suggesting that the formation of other oligomeric complexes might be assisted by chaperones. PMID- 16716594 TI - Gamma-(monophenyl)phosphono glutamate analogues as mechanism-based inhibitors of gamma-glutamyl transpeptidase. AB - Gamma-glutamyl transpeptidase (GGT, EC 2.3.2.2) catalyzes the hydrolysis and transpeptidation of extracellular glutathione and plays a central role in glutathione homeostasis. We report here the synthesis and evaluation of a series of hydrolytically stable gamma-(monophenyl)phosphono glutamate analogues with varying electron-withdrawing para substituents on the leaving group phenols as mechanism-based and transition-state analogue inhibitors of Escherichia coli and human GGTs. The monophenyl phosphonates caused time-dependent and irreversible inhibition of both the E. coli and human enzymes probably by phosphonylating the catalytic Thr residue of the enzyme. The inactivation rate of E. coli GGT was highly dependent on the leaving group ability of phenols with electron withdrawing groups substantially accelerating the rate (Bronsted betalg = -1.4), whereas the inactivation of human GGT was rather slow and almost independent on the nature of the leaving group. The inhibition potency and profiles of the phosphonate analogues were compared to those of acivicin, a classical inhibitor of GGT, suggesting that the phosphonate-based glutamate analogues served as a promising candidate for potent and selective GGT inhibitors. PMID- 16716595 TI - Molecular modeling of non-covalent binding of homochiral (3S,3'S)-astaxanthin to matrix metalloproteinase-13 (MMP-13). AB - Inhibitors for matrix metalloproteinases (MMPs) are under investigation for the treatment of various important chronic illnesses, including cancer, arthritis, and cardiovascular disease (CVD). In particular, MMP-13 is currently being probed as a potential key target in CVD and malignant disease due to its documented effects on extracellular matrix (ECM) remodeling, important in the pathophysiology of these diseases. Within the family of related mammalian MMP enzymes, MMP-13 possesses a large hydrophobic binding pocket relative to that of other MMPs. Homochiral astaxanthin (3S,3'S-AST; 3S,3'S-dihydroxy-beta,beta carotene-4,4'-dione), an important antioxidant and anti-inflammatory xanthophyll carotenoid, is an active metabolite of several novel soft drugs in clinical development; it is also extensively used and tested as a human nutraceutical. In the current study, the prediction of the geometry and energetics of its binding to human MMP-13 was conducted with molecular modeling. The method used was found to predict the energy of binding of known ligands of MMP-13 with great precision. Blind docking using the whole protein target was then used in order to identify the possible binding site(s) of AST. AST was predicted to bind at several sites in close proximity to the active center. Subsequent analyses focused on the binding site at the atomic (i.e., amino acid sequence) level suggested that AST can bind to MMP-13 with high affinity and favorable energetics. Therefore, the modeling study predicts potential direct enzyme-inhibitory activity of AST against MMP-13, a behavior that may be exploited in mammalian systems in which pathological upregulation of MMP activity is paramount. PMID- 16716596 TI - Tyrosinase inhibition studies of cycloartane and cucurbitane glycosides and their structure-activity relationships. AB - In the present paper, tyrosinase inhibition studies and structure-activity relationship of eight cycloartane glycosides and one cucurbitane glycoside and its genin, which were isolated from Astragalus (Leguminoseae) and Bryonia (Cucurbitaceae) plants, have been discussed. The activities are compared with two reference tyrosinase inhibitors, kojic acid and l-mimosine. These studies and the SAR showed that the askendoside B which exhibited highly potent (IC50 =13.95 microM) tyrosinase inhibition could be a possible lead molecule for the development of new medications of several skin diseases related with the over expression of the enzyme tyrosinase, like hyperpigmentation. The molecule also may be interesting for the cosmetic industries as a skin whitening agent. PMID- 16716597 TI - In vivo and in vitro study of porcine oviductal epithelial cells, cumulus oocyte complexes and granulosa cells: a scanning electron microscopy and inverted microscopy study. AB - The morphology and structure of porcine oviductal epithelial cells (POEC), cumulus-oocyte complexes (COCs) and granulosa cells (GC) were investigated in vivo and in vitro conditions using scanning electron microscopy (SEM) and inverted microscopy. The POEC contained columnar ciliated cells and spherical shaped non-ciliated cells. Both non- and ciliated cells appeared either in groups or distributing among each other. However, the isolation of cells was observed after culture for 48 h. A total of 921 oocytes from 20 ovaries was isolated resulting in an average of 46 oocytes per ovary. They were round in shape, surrounded by zona pellucida with layers of cumulus cells ranging between 89.16 and 144.68 microm in size. As for COCs, they were classified into 4 types; intact , multi-, partial-cumulus cell layers and completely denuded oocyte. Interestingly, changes in morphology of COCs with intact and multi-cumulus cell layers were observed in the in vitro study. The GCs in the follicular fluid were also round in shape and found as clusters. After culturing in in vitro for 48 h, no change in morphology was observed. The GC appeared in smaller clusters or were present as single cells and their sizes ranged from 6 to 8 microm. The results obtained from this study allow us to have a better understanding of the morphology and nature of cells under both in vivo and in vitro conditions. This information is also important for the study of their secretions and biochemical compositions, which is of great importance to the use of cells as feeder cells in in vitro fertilization in current studies. PMID- 16716598 TI - Regulation of angiogenesis by hypoxia-inducible factor 1. AB - Hypoxia is an imbalance between oxygen supply and demand that occurs in cancer and in ischemic cardiovascular disease. Hypoxia-inducible factor 1 (HIF-1) was originally identified as the transcription factor that mediates hypoxia-induced erythropoietin expression. More recently, the delineation of molecular mechanisms of angiogenesis has revealed a critical role for HIF-1 in the regulation of angiogenic growth factors. In this review, we discuss the role of HIF-1 in developmental, adaptive and pathological angiogenesis. In addition, potential therapeutic interventions involving modulation of HIF-1 activity in ischemic cardiovascular disease and cancer will be discussed. PMID- 16716600 TI - G-CSF-induced evacuation of sinusoidal NK cells and the facilitation of liver regeneration in a partial hepatectomy. AB - Since liver regeneration after partial hepatectomy (PHx) is known to improve by pretreatment with recombinant human G-CSF (rhG-CSF), we investigated the mechanism by evaluating the distribution and activity of sinusoidal NK cells. F344 rats were treated with rhG-CSF (250 microg/kg/day) for 5 days before PHx. Pretreatment with rhG-CSF improved the serum ALT levels and DNA biosynthesis of the remnant liver tissues at 20 h after PHx. Notably, the rhG-CSF pretreatment decreased the number of NK cells in the liver determined by immunohistochemistry using anti-NKR-P1A mAb before and at 20 h after PHx with no significant change in the NK activity per cell base, while also increasing the number of NK cells in the peripheral blood detected by flow cytometry. The rhG-CSF induced a pre-PHx downregulation of the IL-12p70 protein levels, while also promoting the post-PHx reduction of the protein levels of IL-12p70 and IFN-gamma. Conversely, rhG-CSF had no effect on the pre-PHx mRNA levels or the PHx-induced upregulation of mRNA levels of TNF-alpha, IL-1beta, IL-6, TGF-beta, IL-10, HGF, and c-Met determined by real-time RT-PCR. These results strongly suggest that rhG-CSF-induced facilitation of liver regeneration is achieved by immunoregulation through the intrahepatic IL-12 downregulation and evacuation of sinusoidal NK cells. PMID- 16716602 TI - Expression and purification of the Mycobacterium tuberculosis complex-restricted antigen CFP21 to study its immunoprophylactic potential in mouse model. AB - Secreted proteins encoded by different regions of difference (RDs) from the genome of Mycobacterium tuberculosis have been considered as attractive candidates for vaccination against tuberculosis owing to their absence in most BCG strains. In this study, the structural gene for the RD2 locus encoding protein CFP21 was PCR amplified and expressed as a fusion protein with hexahistidine residues in Escherichia coli. Expression of CFP21 in E. coli under transcriptional regulation of the T7 promoter yielded a protein located within inclusion bodies. The inclusion bodies were solubilized in the presence of 8M urea and the protein was purified to homogeneity under denaturing conditions at low pH using nitrilotriacetic acid (Ni-NTA) affinity chromatography. The denatured protein was renatured by gradient dialysis against a decreasing concentration of urea. The purified protein was shown to have esterase activity. CFP21 protein was evaluated for immunogenicity in C57BL/6J mice. We observed an elevated T cell proliferative response and production of IFN-gamma and IL-12 (p40). CFP21 also induced an optimum level of cytotoxic T cell activity and induced a strong humoral response as indicated by higher levels of specific IgG1 and IgG2a antibody isotypes. In addition, a moderate level of protection was observed against experimental tuberculosis. This is the first report describing esterase activity of the M. tuberculosis complex-restricted protein CFP21 and its protective potential against experimental tuberculosis. PMID- 16716603 TI - Selective neutrality of mitochondrial ND2 sequences, phylogeography and species limits in Sitta europaea. AB - Variation and geographic differentiation in mitochondrial DNA (mtDNA) was studied in the widespread and phenotypically variable Eurasian nuthatch (Sitta europaea). To assess whether sequences were evolving in a selectively neutral fashion, we used McDonald-Kreitman [Nature 351 (1991) 652] tests and a tree-based method, which suggested that although ND2 sequences are affected by natural selection against slightly deleterious alleles, the effects do not compromise phylogeographic inferences. Three phylogenetic species-level clades of nuthatches were discovered, corresponding to the Caucasus, southern Europe, and northern Europe plus Asia. Unimodal mismatch distributions within each clade suggest that populations have undergone recent growth. A westward range expansion was inferred from the geographic pattern in nucleotide diversity. Although samples were insufficient, it is possible that nuthatches in England and Japan are recently differentiated. Two specimens of the subspecies S. e. arctica formed a sister group to all other S. europaea, differing by ca. 10% uncorrected sequence divergence, pointing the need for additional study of this phenotypically distinct taxon. As with other species, mtDNA data support major phenotypic distinctions, but not subspecies. PMID- 16716599 TI - The growth factor independence-1 transcription factor: new functions and new insights. AB - The growth factor independence-1 (Gfi1) transcription factor is required for proper development of neuroendocrine cells, sensory neurons, and blood. Patients with mutations in Gfi1 exhibit severe congenital neutropenia (SCN) or non-immune chronic idiopathic neutropenia of adults. Gfi1 was initially described as an oncoprotein that mediates tumor progression in a mouse model of leukemia; however, recent data suggest that Gfi1 may act as either an oncogene or an anti proliferative tumor suppressor gene depending on the cell type. Here we review the latest literature on Gfi1, and emphasize its role in the hematopoietic, sensory and neuroendocrine systems. PMID- 16716604 TI - Single dose glucagon (0.5 mg IV bolus) administration in healthy human volunteers is a robust model for assessment of glycogenolysis: characterisation of the glucose excursion after glucagon challenge. AB - INTRODUCTION: In the assessment of potential new treatments for Type 2 diabetes, robust pharmacological methods are helpful in assessing efficacy, defining dose response, duration of effect and ultimately in deciding whether to progress compounds to the next phase of drug development. Hepatic glucose handling is abnormal in Type 2 diabetes. We evaluated glucagon challenge as a way of assessing effects on the glycogenolytic pathway. METHODS: In each of 2 studies healthy subjects received glucagon as an IV bolus of 0.5 mg studied after an overnight fast and plasma glucose was monitored before and for 180 min after glucagon challenge. Study 1 was a double-blind placebo controlled study comparing glucagon administered twice with saline placebo. In study 2, subjects were studied on a single occasion and the glucagon challenge was carried out in the morning and then repeated 7 h later. In study 2, insulin concentrations were also monitored before and after the glucagon challenge. RESULTS: In study 1, glucose rose in a reproducible manner with a peak glucose 20 min after challenge falling to baseline values by 120 min and then fell below values for saline challenge between 120 and 180 min. Analysis of the data showed that the corrected AUC(0-20) min was the most robust variable and could be expected to detect clinically relevant changes in small numbers (<10) of subjects. In study 2, we demonstrated that when glucagon challenge was repeated 7 h after the first challenge, the glucose excursion was highly variable. The plasma insulin response was robust following the initial challenge but variable following the second challenge. DISCUSSION: We have demonstrated that an IV bolus glucagon challenge (0.5 mg) results in a reproducible rise in glucose in healthy volunteers and can be repeated within a week but when repeated on the same day gave a poorly reproducible rise in glucose. Glucagon challenge may be useful in studying novel drugs that affect glycogen handling in the liver. PMID- 16716605 TI - MRI of articular cartilage in OA: novel pulse sequences and compositional/functional markers. AB - Osteoarthritis (OA) is a leading cause of disability worldwide. Magnetic resonance imaging (MRI), with its unique ability to image and characterize soft tissue non-invasively, has proven valuable in assessing cartilage in OA. The development of new, fast imaging methods with high contrast show promise to improve the magnetic resonance (MR) evaluation of this disease. In addition to morphologic MRI methods, MRI contrast mechanisms under development may reveal detailed information about the physiology of cartilage. It is anticipated that these and other MRI techniques will play an increasingly important role in assessing the success or failure of therapies for OA. On December 5 and 6, 2002, OMERACT (Outcome Measures in Rheumatology Clinical Trials) and OARSI (Osteoarthritis Research Society International) held a workshop in Bethesda, MD aiming at providing a state-of-the-art review of imaging outcome measures for OA of the knee to help guide scientists and pharmaceutical companies in the use of MRI in multi-site studies of OA. Applications of MRI were initially reviewed by a multidisciplinary, international panel of expert scientists and physicians from academia, the pharmaceutical industry and regulatory agencies. The findings of the panel were then presented to a wider group of participants for open discussion. The following report summarizes the results of these discussions with respect to novel MRI pulse sequences for evaluating articular cartilage of the knee in OA and notes any additional advances that have been made since. PMID- 16716606 TI - The effect of tibial lengthening on immature articular cartilage of the knee joint. AB - OBJECTIVE: To investigate the acute response of immature articular cartilage, in the distraction and consolidation phases, to 30% tibial lengthening. DESIGN: Sixteen immature New Zealand white rabbits underwent diaphyseal lengthening of the left tibia by callotasis at a distraction rate of 0.4mm twice daily. A sham control group of 12 rabbits underwent fixation and osteotomy without lengthening. In each group, half of the rabbits were killed at the end of the distraction phase or at an equivalent time period and the rest were killed after an additional 5 weeks (consolidation phase). The tibial condyles and synovial fluid in the knee joint cavity were taken for laboratory examination. Sulfated glycosaminoglycan in synovial fluid was estimated using a colorimetric method. Sections along the mid-coronal plane of the whole of the tibial condyles were examined histologically and by scanning electron microscopy. A grading system was used to make a semiquantitative assessment of the histopathological changes in articular cartilage. RESULTS: Damage to the immature articular cartilage had occurred by the end of the distraction period and the cartilage continued to deteriorate in the consolidation phase of 5 weeks. However, when compared with a similar study in a mature rabbit model, damage to immature cartilage appeared less severe. CONCLUSION: This model of 30% lengthening caused acute cartilage damage which did not recover in the short term. The result may have implications for longer lengthening in children; the effects may be disadvantageous and lead to degenerative diseases later in life. PMID- 16716607 TI - Differentiating characterization of human umbilical cord blood-derived mesenchymal stem cells in vitro. AB - It has been demonstrated that the number and differentiating potential of bone marrow mesenchymal stem cells (MSCs) decrease with age. Therefore, the search for alternative sources of MSCs is of significant value. In the present study, MSCs were isolated from umbilical cord blood (UCB) by combining gradient density centrifugation with plastic adherence. Cultured cells were treated with ascorbate acid-2-phosphate, dexamethasone, beta-glycerophosphate dexamethasone, insulin, 1 methyl-3-isobutylxamthine, indomethacin, beta-mercaptoethanol, butylated hydroxyanisole, FGF-4 and HGF. Differentiating characterization of UCB-derived MSCs were detected by cytochemistry, immunocytochemistry, radioimmunoassay, RT PCR and urea assay. The results showed UCB-derived MSCs could differentiate into osteoblasts, adipocytes and neuron-like cells. When MSCs were cultured with FGF-4 and HGF, approximately 63.6% of cells became small, round and epithelioid on day 28 by morphology. Compared with the control, levels of AFP in the supernatant liquid increased significantly from day 12 and were higher on day 28 (P<0.01). Albumin increased significantly from day 16 (P<0.01). Urea was first detected on day 20 (P<0.01), and continued to increase on day 28 (P<0.01). Cells first expressed CK-18 on day 16 through immunocytochemistry analysis. RT-PCR analysis showed that differentiated cells could express a number of hepatocyte-specific genes in a time-dependent manner. Glycogen storage was first seen on day 24. Our results suggest that UCB-derived MSCs can differentiate not only into osteoblasts, adipocytes and neuron-like cells, but also into hepatocytes. Human UCB-derived MSCs are a new source of cell types for cell transplantation and therapy. PMID- 16716608 TI - Morphological and growth alterations in Vero cells transformed by cisplatin. AB - Cisplatin is an antineoplastic agent used to treat solid tumours, such as ovarian, testicular and bladder tumours. However, studies in vitro and in vivo have shown that cisplatin is mutagenic, genotoxic and tumorigenic in other tissues and organs. In this work, we examined the effect of cisplatin on Vero cells, a fibroblast-like cell line. The morphological characteristics were investigated using phase contrast microscopy, scanning electron microscopy and the actin cytoskeleton was labelled with fluorescein isothiocyanate-phalloidin. Cell proliferation was assessed based on the growth curve. Cultured Vero cells treated with cisplatin showed behavioural and morphological alterations associated with cellular transformation. The transformed cells grew in multilayers and formed cellular aggregates. The proliferation and morphological characteristics of the transformed cells were very different from those of control ones. Since transformed Vero cells showed several characteristics related to neoplastic growth, these cells could be a useful model for studying tumour cells in vitro. PMID- 16716609 TI - Effects of Ginkgolide on the development of NOS and AChE positive neurons in the embryonic basal forebrain. AB - Extract of Ginkgo biloba (EGb) has been therapeutically used for several decades to increase peripheral and cerebral blood flow so as to prevent cardiovascular and neurovascular diseases. However, the role of EGb in neuroprotective effects has received much attention recently. In this study, we investigated the effect of EGb on the development of NOS and AChE positive neurons in the rat embryonic basal forebrain. The results showed that treated with EGb, the OD of MTT staining analysis, and the numbers, the cell sizes and circumferences of NOS and AChE positive neurons were greatly promoted. These data suggest that EGb had similar effects of the neurotrophins such as NGF and BDNF in promoting the development of NOS and AChE positive neurons in the rat embryonic basal forebrain. PMID- 16716610 TI - Prevalence of MRSA in emergency and elective patients admitted to a vascular surgical unit: implications for antibiotic prophylaxis. AB - OBJECTIVES: MATERIALS AND METHODS: 200 consecutive emergency/transfer and 150 consecutive elective patients admitted between April 2004 and January 2005, were studied. Data was obtained from departmental Morbidity and Mortality records and the computerised laboratory medicine information system. RESULTS: 261 (75%) of the 350 patients were screened for MRSA on admission (target 100%). The proportions of emergency/transfer and elective patients screened were similar (78% and 72% respectively). The prevalence of MRSA carriage detected by admission screening in emergency/transfer patients 30/153 (20%), was significantly higher (p<0.0001) than in elective patients 2/108 (2%). A simple decision analysis model suggests that gentamicin should be used when the prevalence of MRSA reaches 10% and vancomycin when the prevalence reaches 50%. CONCLUSIONS: The high prevalence of MRSA colonisation in emergency/transfer patients has important implications for pre-operative antibiotic prophylaxis. PMID- 16716611 TI - Early carotid endarterectomy after ischemic stroke: the results of a prospective multicenter Italian study. PMID- 16716612 TI - A modified technique of delayed basilic transposition - initial results. AB - The radiocephalic wrist fistula remains the "gold standard" for primary vascular access placement according to NKF -DOQI guidelines. However, liberal inclusion criteria for dialysis therapy, as well as increasing longevity and prevalence of diabetes, have led to an increased number of patients with distal vessels unsuitable for vascular access creation. Another available location, and one used with increasing frequency, is the anterior cubital fossa. Arteriovenous anastomoses in this location leads to the development of two possible vascular accesses: cephalic and basilic. The basilic vein needs to be transposed to the anterior aspect of the arm. However, this procedure is technically difficult and carries a high risk of complications, often associated with axial torsion or kinking of the transposed vein during placement in the subcutaneous tunnel. The authors propose a simple technical modification which minimizes the risk of these complications. The main advantage is the prevention of axial torsion and kinking, as well as avoiding venous stenosis by transecting the vein at its widest point. PMID- 16716613 TI - Prognostic significance of raised cardiac troponin T in patients presenting with acute limb ischaemia. AB - OBJECTIVE: To study the relation between serum cardiac troponin T (cTnT) and mortality in patients presenting with acute limb ischaemia secondary to an embolism. MATERIAL AND METHODS: A two years prospective study of all patients admitted to the vascular unit with a diagnosis of acute limb ischaemia secondary to an embolism. On admission all patients had an ECG. A blood sample was taken for measurement of cTnT, CRP, serum biochemistry, full blood count and clotting. All embolectomies were performed under local anaesthesia. Patients were followed until discharge from hospital and up to twelve months after surgery. RESULTS: There were 37 patients with lower limb and 2 patients with upper limb ischaemia. Twenty four patients were female and fifteen were male, with the mean age of 76 years (50-95) for women and 84 years (77-90) for men. Seventeen patients (44%) had a raised cTnT. The patients with raised cTnT were older than those with normal cTnT [86y (77-92) vs 77y (51-95), p = 0.01, t test]. The mean cTnT was 0.20 microg/L (range: 0.11-0.27). Only two patients with raised cTnT gave a history of chest pains. All of the patients with an elevated cTnT had also raised CRP. There was no significant difference in the serum creatinine in the group of patients with elevated cTnT compared to those with normal cTnT [112 micromol/L (range 98-159) vs 119 micromol/L (range: 47-177), p = ns]. The cumulative survival for cTnT+ patients at 7 days was 53% and that of cTnT- patients was 100%. The cumulative survival for cTnT+ and cTnT- patients was statistically different (p = 0.0000, chi2 = 13.1, Log Rank test). Using regression analysis, an elevated cTnT was found to be an independent predictor of outcome. CONCLUSION: A significant proportion of patients presenting with an acutely ischaemic limb have an elevated cTnT. An elevated cTnT may be an early marker of overall disease severity and a predictor of outcome. PMID- 16716614 TI - Occult renal impairment is common in patients with peripheral vascular disease and normal serum creatinine. AB - OBJECTIVE: The incidence of peripheral vascular disease (PVD) and angiography/angioplasty is rising annually. The UK Small Aneurysm Trial and other trials have shown renal function is a predictor of increased mortality and failed infrainguinal bypass despite patent vessels. Renal function is classically assessed by serum creatinine (SCr). However, SCr can be normal despite significant renal impairment. A more sensitive test is creatinine clearance (CrCl) as determined by 24-hour urine collection in combination with SCr. We studied the incidence of renal impairment, as defined by CrCl, in PVD patients with normal SCr. METHODOLOGY: All patients with PVD sufficient to necessitate angiography and normal SCr (< or =120 micromol/l - men; < or =97 micromol/l - women) had their CrCl assessed prior to angiography: using both 24-hour urine collection and the Cockcroft-Gault formula. Various blood tests, a detailed history and examination were performed. A control group of arthritic patients, age and sex-matched with similar SCr, also had their CrCl determined. RESULTS: 65 of 76 patients (86%) with normal SCr had a subnormal CrCl (<100 ml/min) and 49 (65%) had a CrCl below 60 ml/min. In the control group of arthritic patients, the proportion having impaired CrCl was significantly less - 67% below 100 mls/min (p=0.0471) and only 15% below 60 mls/min (p<0.0001). The median and interquartile range CrCl of 52 [38-81] mls/min for PVD patients was significantly worse than for control patients (80 [68-119] mls/min -p<0.0001). The Cockcroft-Gault formula for calculating CrCl did not correlate well with the urinary CrCl for the control group but did for PVD patients (p<0.0001). Factors associated with a significantly reduced CrCl were age of at least 75 years, SCr of at least 85 micromol/l and a history of coronary heart disease (all p<0.05). This had a sensitivity of 88% and specificity of 82% for identifying subnormal CrCl. Statin use was associated with a significantly improved CrCl (p=0.040). CONCLUSION: Most PVD patients with normal serum creatinine have occult, significantly impaired renal function as defined by creatinine clearance. Vascular surgeons should include creatinine clearance in pre-operative assessment of renal function especially in patients over 75 years old, with a history of coronary heart disease or a serum creatinine over 85 micromol/l. The method of determining creatinine clearance could be the Cockcroft-Gault calculation or ideally 24-hour urinary creatinine clearance measurement. This would allow appropriate early referral to a nephrologist for further investigation and management. It is worth noting that statin use seems to be associated with a protective effect on renal function. PMID- 16716615 TI - Continuous brachial plexus blockade in combination with the NMDA receptor antagonist memantine prevents phantom pain in acute traumatic upper limb amputees. AB - BACKGROUND: Hyperexcitability of N-methyl-d-aspartate acid (NMDA) receptors may play an important role in the development of phantom limb pain (PLP). AIM OF THE STUDY: To investigate whether early treatment with the NMDA antagonist memantine attenuates phantom pain memory formation in traumatic amputees. METHODS: In a randomized, double-blind, controlled trial 19 patients with acute traumatic amputation of the upper extremity were investigated. All patients received postoperative analgesia by continuous brachial plexus anesthesia (ropivacaine 0.375% 5 ml/h) for at least 7 days. In addition, the patients received either memantine (20-30 mg daily, n=10) or placebo (n=9) for 4 weeks. RESULTS: Memantine treatment reduced the number of requested ropivacacine bolus injections during the first week and resulted in a significant decrease of PLP prevalence and intensity at 4 weeks and 6 months follow up, but not at 12 months follow up. CONCLUSIONS: We conclude that memantine can reduce intensity of phantom limb pain and might also prevent the development of PLP. However, despite the very early begin of treatment; no long-term effect on established PLP was evident. PMID- 16716616 TI - Practical design of a 4 Tesla double-tuned RF surface coil for interleaved 1H and 23Na MRI of rat brain. AB - MRI is proving to be a very useful tool for sodium quantification in animal models of stroke, ischemia, and cancer. In this work, we present the practical design of a dual-frequency RF surface coil that provides (1)H and (23)Na images of the rat head at 4 T. The dual-frequency RF surface coil comprised of a large loop tuned to the (1)H frequency and a smaller co-planar loop tuned to the (23)Na frequency. The mutual coupling between the two loops was eliminated by the use of a trap circuit inserted in the smaller coil. This independent-loop design was versatile since it enabled a separate optimisation of the sensitivity and RF field distributions of the two coils. To allow for an easy extension of this simple double-tuned coil design to other frequencies (nuclei) and dimensions, we describe in detail the practical aspects of the workbench design and MRI testing using a phantom that mimics in vivo conditions. A comparison between our independent-loop, double-tuned coil and a single-tuned (23)Na coil of equal size obtained with a phantom matching in vivo conditions, showed a reduction of the (23)Na sensitivity (about 28 %) because of signal losses in the trap inductance. Typical congruent (1)H and (23)Na rat brain images showing good SNR ((23)Na: brain 7, ventricular cerebrospinal fluid 11) and spatial resolution ((23)Na: 1.25 x 1.25 x 5mm(3)) are also reported. The in vivo SNR values obtained with this coil were comparable to, if not better than, other contemporary designs in the literature. PMID- 16716617 TI - Cell adhesion characteristics of a monocytic cell line derived from rainbow trout, Oncorhynchus mykiss. AB - In experiments investigating the adhesive properties of the rainbow trout splenic monocyte-like cell line RTS11 it was found that the cells bound with low affinity to plates coated with bovine serum albumin (BSA) but that phorbol ester-induced activation/differentiation greatly increased adhesion to BSA. Similarly, pre exposure to 500 microM MnCl(2) at time of plating, increased RTS11 adhesion to BSA coated plates, in agreement with the reported ability of divalent cations such as Mn(2+) to activate integrins. Integrins are a diverse family of heterodimeric cell surface glycoproteins that have been shown to mediate cell cell and cell-extracellular matrix adhesion. Transcripts of the beta(2)-integrin CD18 were detected by PCR in RTS11 but not in RTG-2 cells, a fibroblastic lineage derived from rainbow trout gonads. These results suggest that differentiated RTS11 express molecules related to members of the beta(2)-integrin subfamily such as the macrophage lineage marker Mac-1 (CD11b/CD18) and/or p150,95 (CD11c/CD18) and possibly as well alpha(4)beta(1) of the beta(1)-integrin subfamily. PMID- 16716618 TI - Correlation of seasonal acclimatization in metabolic enzyme activity with preferred body temperature in the Eastern red spotted newt (Notophthalmus viridescens viridescens). AB - Eastern red spotted newts, as aquatic adults, are active year round. They are small and easy to handle, and thus lent themselves to a laboratory study of seasonal changes in preferred body temperature and biochemical acclimatization. We collected newts in summer (n=20), late fall (n=10) and winter (n=5). Ten each of the summer and late fall newts were subjected to an aquatic thermal gradient. Summer newts maintained higher cloacal temperatures than late fall newts (26.8+/ 0.5 degrees C and 17.2+/-0.4 degrees C, respectively). In addition, the activity of three muscle metabolic enzymes (cytochrome c oxidase (CCO), citrate synthase (CS) and lactate dehydrogenase (LDH)) was studied in all newts collected. Newts compensated for lower late fall and winter temperatures by increasing the activity of CCO during those seasons over that in summer newts at all assay temperatures (8, 16 and 26 degrees C). The activity of CS was greater in winter over summer newts at 8 and 16 degrees C. No seasonal differences in LDH activity were demonstrated. These data in newts indicate that this amphibian modifies some muscle metabolic enzymes in relation to seasonal changes and can modify its behavioral in a way that correlates with those biochemical changes. PMID- 16716619 TI - Effects of temperature and calcium availability on cardiac contractility in Synbranchus marmoratus, a neotropical teleost. AB - An isometric muscle preparation was used to investigate the importance of the ventricular sarcoplasmic reticulum (SR) and extracellular Ca2+ (1.25 up to 11.25 mM) to force generation at 25 degrees C (acclimation temperature), 15 and 35 degrees C. The post-rest tension and force-frequency relationship were conducted with and without 10 microM ryanodine in the bathing medium. Increments in extracellular Ca2+ resulted in increases in twitch force development only at 35 degrees C. A significant post-rest potentiation was recorded for the control preparations at 25 degrees C (100% to 119.8+/-4.1%). However, this post-rest potentiation was inhibited by ryanodine only at 25 degrees C (100% to 97.6+/ 1.5%). At 35 degrees C, force remained unchanged in the control preparations, but a significant post-rest decay was recorded in the presence of ryanodine (100% to 76.6+/-4.6%) while at 15 degrees C, ryanodine was not able to preventing the post rest potentiation observed in the control preparations. The increases in the imposed contraction frequency caused a decline of the force at 25 and 35 degrees C and ryanodine decreased significantly peak tension at both temperatures. The findings suggest a high or medium calcium turnover, possibly related to the presence of a functional SR, whose functionality is diminished when temperature is decreased. PMID- 16716620 TI - Do sperm contribute to the buffering capacity of steelhead trout (Oncorhynchus mykiss) semen? AB - The motility of salmonid sperm is pH-sensitive and the buffering capacity of the seminal plasma is low. The objective of the present study was to determine the extent to which sperm contribute to the buffering capacity of whole steelhead (Oncorhynchus mykiss) semen. To determine the buffering capacity, semen and seminal plasma samples were titrated with HCl and pH measurements taken at 1-2 min. The buffering capacity of semen was not different from that of seminal plasma over the pH range 7.5 to 8.5 and was approximately 15% to 20% less over the range 6.0 to 7.0. Comparable results were obtained for the semen and seminal plasma of the chinook salmon (Oncorhynchus tshawytscha). To assess whether the intracellular environment could influence the buffering capacity, the effects of cell disruption with n-butanol and Triton X-100 (TX-100) were determined. Over the pH range 7.5 to 8.5, the presence of n-butanol or TX-100 resulted in a doubling of the buffering capacity of the semen; TX-100, but not n-butanol, increased semen buffering capacity over the pH range 6.0 to 7.0. To determine whether the sperm's intracellular compartment might contribute to the buffering capacity over a longer duration, semen and seminal plasma samples were acidified with HCl and the pH measured over several hours. These data suggest that intact sperm contribute no more than about 25% to the buffering capacity of whole semen. The buffering capacity of steelhead semen and seminal plasma were comparably and modestly temperature sensitive. The results suggest that the sperm may contribute to the buffering capacity of the semen over a physiological pH range, however, if so, the effect is relatively small. PMID- 16716621 TI - Specific dynamic action: a century of investigation. AB - Specific dynamic action (SDA) is the term used to refer to the increased metabolic expenditure that occurs in postprandial animals. Postprandial increases in metabolism were first documented in animals over two hundred years ago, and have since been observed in every species thus far examined. Ironically, the ubiquity of this physiological response to feeding understates its complex nature. This review is designed to summarize both classical and modern hypotheses regarding the causality of SDA as well as to review important findings from the past century of scientific research into SDA. A secondary aim of this work is to emphasize the importance of carefully designed experiments and systematic hypothesis testing to make more rapid progress in understanding the physiological processes that contribute to SDA. I also identify three areas in SDA research that deserve more detailed investigation. The first area is identification of the causality of SDA in 'model' organisms. The second area is characterization of SDA responses in novel species. The third area is exploration of the ecological and potential evolutionary significance of SDA in energy budgets of animals. PMID- 16716622 TI - Evolution of GnRH ligands and receptors in gnathostomata. AB - Gonadotropin-releasing hormone (GnRH) is the final common signaling molecule used by the brain to regulate reproduction in all vertebrates. Until now, a total of 24 GnRH structural variants have been characterized from vertebrate, protochordate and invertebrate nervous tissue. Almost all vertebrates already investigated have at least two GnRH forms coexisting in the central nervous system. Furthermore, it is now well accepted that three GnRH forms are present both in early and late evolved teleostean fishes. The number and taxonomic distribution of the different GnRH variants also raise questions about the phylogenetic relationships between them. Most of the GnRH phylogenetic analyses are in agreement with the widely accepted idea that the GnRH family can be divided into three main groups. However, the examination of the gnathostome GnRH phylogenetic relationships clearly shows the existence of two main paralogous GnRH lineages: the ''midbrain GnRH" group and the "forebrain GnRH" group. The first one, represented by chicken GnRH-II forms, and the second one composed of two paralogous lineages, the salmon GnRH cluster (only represented in teleostean fish species) and the hypophysotropic GnRH cluster, also present in tetrapods. This analysis suggests that the two forebrain clades share a common precursor and reinforces the idea that the salmon GnRH branch has originated from a duplication of the hypophysotropic lineage. GnRH ligands exert their activity through G protein-coupled receptors of the rhodopsin-like family. As with the ligands, multiple GnRHRs are expressed in individual vertebrate species and phylogenetic analyses have revealed that all vertebrate GnRHRs cluster into three main receptor types. However, new data and a new phylogenetic analysis propose a two GnRHR type model, in which different rounds of gene duplications may have occurred in different groups within each lineage. PMID- 16716623 TI - Adaptations to fasting in a terrestrial mustelid, the sable (Martes zibellina). AB - The aim of this study was to investigate whether the actively wintering Palearctic sable Martes zibellina has evolved physiological adaptations to tolerate nutritional scarcity. Sixteen farm-bred male sables were divided into a fed control group and an experimental group fasted for 4 days. The rate of weight loss in the sable was similar to other medium-sized mustelids. Fasting led to hypoglycaemia and to a decreased lymphocyte percentage. The sable derived metabolic energy from both subcutaneous and intraabdominal white adipose tissues and the relative decrease in fat mass was the largest for the retroperitoneal and subcutaneous depots. Metabolic energy derived partly from body proteins indicated by the increased plasma levels of urea, uric acid and total essential amino acids. Triacylglycerols accumulated in the livers of the fasted sables and the increased plasma aminotransferase activities suggested hepatic dysfunction. The decreased plasma insulin concentrations and the elevated cortisol levels probably contributed to stimulated lipolysis and protein catabolism. Moreover, fasting increased the plasma ghrelin concentrations of the sables and down-regulated the thyroid activity. PMID- 16716624 TI - Influence of diets supplemented with vitamins C and E on pirarucu (Arapaima gigas) blood parameters. AB - This study evaluated the influence of diets supplemented with 500, 800, 1200 mg kg-1 of vitamin C (ascorbic acid or AA) and vitamin E (alpha-tocopherol or alpha T) on the physiological responses of pirarucu fed for 2 months. Weight and mortality were not affected by dietary vitamin type or their concentrations. Significant increase (p<0.05) on the red blood cells count was obtained on treatments with 800 and 1200 mg AA kg-1 and on the hemoglobin concentration on treatment with 500 mg alpha-T kg-1 relatively to control. Mean corpuscular volume presented a significant decrease (p<0.05) on treatment with 800 and 1200 mg AA kg 1 when compared to control. Mean corpuscular hemoglobin concentration was significantly high (p<0.05) on treatment with 500 mg alpha-T kg-1. Only in vitamin C treatments, we noticed a significant increase (p<0.05) in the number of leucocytes relative to control. All fish in the vitamin-supplemented treatments, except 500 mg AA kg-1, had high total protein values compared to control. Fish treated with 800 or 1200 mg alpha-T kg-1 also showed increases in plasma glucose concentrations. Our results suggest that 800 and 1200 mg AA kg-1 are probably the most suitable concentrations for pirarucu diets, although high vitamin E diets are not necessary for quantitative leucocyte increases for this species. PMID- 16716625 TI - A second form of collagenous lectin from the tunicate, Styela plicata. AB - This study characterised a 90 kDa lectin from an invertebrate chordate, the tunicate Styela plicata. One- and two-dimensional electrophoresis showed that the apparent molecular weight of this protein is maintained under both reducing and non-reducing conditions, suggesting that its native form is a monomer. The 90 kDa lectin was localised within a single type of hemocyte (morula cells), but was secreted from those cells when tunicates were challenged with the inflammatory elicitor, zymosan. Functional studies showed that the 90 kDa protein binds to galactose-based sugars in a divalent cation-dependent manner. Amino acid composition analysis and N-terminal amino acid sequencing indicated that the 90 kDa lectin is related to a previously characterised, collagenous lectin from S. plicata, splic43. However, peptide mass fingerprinting identified numerous differences between the two proteins. This suggests that the 90 kDa molecule represents a novel protein that is involved in host defence. PMID- 16716626 TI - Insights into the substrate specificity of a novel snake venom serine peptidase by molecular modeling. AB - The cDNA encoding BthaTL, a serine peptidase from the venom of the snake Bothrops alternatus, was cloned and sequenced. The deduced primary structure shows over 62% of identity with snake venom thrombin-like enzymes (SVTLEs), molecules with high substrate specificity toward different natural substrates. Indeed, a phylogenetic reconstruction by two different methods clustered this enzyme close to other SVTLEs. These enzymes generally affect the hemostatic system in several ways, and therefore are used as tools in pharmacology and clinical diagnosis. A three-dimensional model of BthaTL was built by homology modeling using TSV-PA (Trimeresurus stejnegeri venom plasminogen activator) crystal structure as template. BthaTL model showed that the typical catalytic triad conformation of serine peptidases was preserved. The calcium coordination ligands were absent or adopt an unfavorable conformation, preventing interactions with metals. On the other hand, the Asp97-Arg174 saline bridge of TSV-PA was not found and its specificity determinant Phe193 is replaced by a Gly in BthaTL. The substitution of essential residues in the neighborhoods of the catalytic site cleft of BthaTL indicates that these two proteins do not share the same enzymatic specificity, what means that BthaTL will probably not activate plasminogen. Such observations may be helpful in the understanding of the molecular mechanism for substrate specificity of these enzymes. PMID- 16716627 TI - Effects of insulin, triiodothyronine and fat soluble vitamins on adipocyte differentiation and LPL gene expression in the stromal-vascular cells of red sea bream, Pagrus major. AB - Various kinds of hormones including insulin, triiodothyronine (T(3)) and fat soluble vitamins have been proposed as mediators of adipocyte differentiation in mammals. To investigate the factors which are responsible for fish adipocyte differentiation, we developed a serum-free culture system of stromal-vascular cells of red sea bream adipose tissue and examined the effects of bovine insulin, T(3), and fat-soluble vitamins (all-trans retinoic acid, retinyl acetate and 1,25 dihydroxyvitamin D(3)) on the differentiation-linked expression of the lipoprotein lipase (LPL) gene. As assessed by the increase in LPL gene expression after 3 day cultivation, like in mammalian adipocytes, insulin enhanced the adipocyte differentiation in a concentration-dependent manner. During 2 week cultivation, bovine insulin promoted lipid accumulation in differentiating adipocytes concentration-dependently until the terminal differentiation. These results indicate that the differentiation of fish adipocytes is inducible by insulin alone. T(3) alone had no effect but enhanced the differentiation-linked LPL gene expression in the presence of insulin. Fat-soluble vitamins, unlike in mammalian adipocytes, did not show any significant effects. The method developed in this study should be of interest for the characterization of factors involved in fish adipocyte differentiation. PMID- 16716628 TI - C-type natriuretic peptide in growth: a new paradigm. AB - C-type natriuretic peptide (CNP), acting through its receptor, natriuretic peptide receptor-B (NPR-B), plays a critical role in linear growth. Knockout mice for CNP and NPR-B are dwarfed, and transgenic mice overexpressing CNP are overgrown. CNP has a direct regulatory effect on growth plate chondrocytes, acting primarily to promote terminal differentiation and hypertrophy. In humans, homozygous NPR-B mutations are the cause of acromesomelic dysplasia, Maroteaux type (AMDM), a severe form of disproportionate dwarfism. A patient with AMDM and the NPR-B knockout mouse both have low insulin-like growth factor I (IGF-I) levels, suggesting an interaction between these regulatory systems. Heterozygous carriers of NPR-B mutations also have reduced stature, but no other abnormalities. Hence, heterozygous NPR-B mutations are another cause of "idiopathic" short stature. The CNP-NPR-B system has only recently been found to be an important regulator of human growth, and abnormalities in this system have clinical implications. Considerable work is needed to further understand this new paradigm of human growth regulation. PMID- 16716631 TI - Genes and mutations conferring antimicrobial resistance in Salmonella: an update. AB - Resistance to various classes of antimicrobial agents has been encountered in many bacteria of medical and veterinary relevance. Particular attention has been paid to zoonotic bacteria such as Salmonella. Over the years, various studies have reported the presence of genes and mutations conferring resistance to antimicrobial agents in Salmonella isolates. This review is intended to provide an update on what is currently known about the genetic basis of antimicrobial resistance in Salmonella. PMID- 16716629 TI - Rskalpha-actin/hIGF-1 transgenic mice with increased IGF-I in skeletal muscle and blood: impact on regeneration, denervation and muscular dystrophy. AB - Human IGF-I was over-expressed in skeletal muscles of C57/BL6xCBA mice under the control of the rat skeletal alpha-actin gene promoter. RT-PCR verified expression of the transgene in skeletal muscle but not in the liver of 1- and 21-day old heterozygote transgenic mice. The concentration of endogenous mouse IGF-I, measured by an immunoassay which does not detect human IGF-I, was not significantly different between transgenic mice and wild-type littermates (9.5 +/ 0.8 and 13.3 +/- 1.9 ng/g in muscle; 158.3 +/- 18.6 and 132.9 +/- 33.1 ng/ml in plasma, respectively). In contrast, quantitation with antibodies to human IGF-I showed an increase in IGF-I of about 100 ng/ml in plasma and 150 ng/g in muscle of transgenic mice at 6 months of age. Transgenic males, compared to their age matched wild-type littermates, had a significantly higher body weight (38.6 +/- 0.53 g vs. 35.8 +/- 0.64 g at 6 months of age; P < 0.001), dry fat-free carcass mass (5.51 +/- 0.085 vs. 5.08 +/- 0.092 g; P < 0.001) and myofibrillar protein mass (1.62 +/- 0.045 vs. 1.49 +/- 0.048 g; P < 0.05), although the fractional content of fat in the carcass was lower (167 +/- 7.0 vs. 197 +/- 7.7 g/kg wet weight) in transgenic animals. There was no evidence of muscle hypertrophy and no change in the proportion of slow type I myofibres in the limb muscles of Rskalpha actin/hIGF-I transgenic mice at 3 or 6 months of age. Phenotypic changes in Rskalpha-actin/hIGF-I mice are likely to be due to systemic as well as autocrine/paracrine effects of overproduction of IGF-I due to expression of the human IGF-I transgene. The effect of muscle specific over-expression of Rskalpha actin/hIGF-I transgene was tested on: (i) muscle regeneration in auto transplanted whole muscle grafts; (ii) myofibre atrophy following sciatic nerve transection; and (iii) sarolemmal damage and myofibre necrosis in dystrophic mdx muscle. No beneficial effect of muscle specific over-expression of Rskalpha actin/hIGF-I transgene was seen in these three experimental models. PMID- 16716632 TI - The epidemiology of antibiotic resistance in Campylobacter. AB - Antibiotic resistance, particularly with the fluoroquinolones and macrolide antibiotics, has now emerged globally with thermophilic campylobacters, including Campylobacter jejuni and C. coli, giving rise to concerns about how these organisms have acquired such resistance characteristics, as well as consequences for human and animal treatment. This review examines (i) the clinical epidemiology of antibiotic resistance in human and animal thermophilic campylobacters, (ii) an update on resistance rates globally, (iii) surveillance of antimicrobial resistance in campylobacters originating from animals, particularly poultry, (iv) the role of the environment in the acquisition and transmission of antibiotic-resistant campylobacters, as well as (v) issues of biocide resistance in campylobacters. PMID- 16716633 TI - Antimicrobial resistance islands: resistance gene clusters in Salmonella chromosome and plasmids. AB - Genes conferring simultaneous resistance to different classes of antimicrobials, confer a selective advantage to the host, particularly when those corresponding antibiotics are administered. Multiple resistance genes clustered within the same genetic locus (resistance island) can be transferred en bloc to other organisms. In this chapter we review novel multidrug resistance islands recently described in Salmonella. PMID- 16716634 TI - [Assisted Reproductive Techniques: SOS!]. PMID- 16716635 TI - [Proposition of a simplified investigation model for adverse outcomes in obstetrics and gynaecology]. AB - Investigation of adverse outcomes in obstetrics and gynaecology requires a systematic and systemically oriented approach. The main objective of these assessments is to decrease their incidence. Lethal complications in obstetrics and gynaecological surgery have become so rare in developed countries that we have to grapple mainly with severe morbidities. A model for such investigation is presented, based on answers to 3 questions: What? How? and Why? These questions correspond to 3 components of quality assurance: Outcome, Process, and Structure, respectively. Aspects of Reason's model of accidents are also included, in the form of "active failures" and "latent conditions." The latter are divided into basic contributory factors, 4 of which are derived from a model of human factors influencing behaviour in anaesthesia or flight-crew environments. To illustrate its basic nature, the model is applied to anonymized cases of surgical complications. Application of the model to the reactive investigation of close calls is discussed, as is application to the proactive interrogation of the care system for the presence and nature of hazards that pose threats to safety. PMID- 16716636 TI - [Prenatal screening for Down syndrome: from sequential to combined estimated risk]. PMID- 16716637 TI - Biomedical sensor using thick film technology for transcutaneous oxygen measurement. AB - Transcutaneous blood gas monitoring is a non-invasive measurement technique for obtaining fast and relatively accurate responses to determine the respiratory conditions of patients. In this investigation, a screen-printed, disposable, transcutaneous oxygen sensor based on the working principle of amperometry, incorporates an integral heating element to enhance transcutaneous diffusion of blood gases typically at 44 degrees C. A Clark cell configuration is employed with gold working and counter electrodes and a silver/silver chloride reference electrode. Two different types of electrolytes namely potassium nitrate gel and polyelectrolyte (Nafion) have been studied under laboratory test conditions. A fully computer-controlled gas testing rig has been constructed to automate the varying of oxygen levels. Linear relationships have been established with an averaged sensitivity level of 0.029 microA/mmHg. In addition, a brief pilot clinical trial involving a fully grown human subject has been carried out alongside a commercial transcutaneous blood gas analyser. The investigations have shown that although the measured signals are weaker than those obtained from the laboratory test, the thick film sensor displays a repeatable and linear relationship when correlating with the commercial system. This study has greatly contributed towards the understanding for the suitability of the materials in achieving a viable, low-cost biomedical sensor. PMID- 16716638 TI - Optimal cut of trabecular network. AB - It has been shown recently that failure of mechanically tested trabecular bone samples can be localized within a part of the volume of the samples. Bone volume fraction BV/TV of failure regions was found to be a better determinant of the mechanical competence of the specimens and was smaller than BV/TV of the whole samples. These results suggest that localization of a failure within a part of an inhomogeneous trabecular network can be related to the presence of a surface of minimal cut-a surface separating the analyzed trabecular sample into two disjoint parts in such a way, that the separation requires removal of minimal possible amount of bone material. Thus, to properly address the problem of mechanical competence of a trabecular bone sample, one must be able to detect and describe the surface of minimal cut. In this paper an algorithm localizing surfaces of minimal cut within 3D trabecular structures is introduced. PMID- 16716639 TI - Retinal assessment using optical coherence tomography. AB - Over the 15 years since the original description, optical coherence tomography (OCT) has become one of the key diagnostic technologies in the ophthalmic subspecialty areas of retinal diseases and glaucoma. The reason for the widespread adoption of this technology originates from at least two properties of the OCT results: on the one hand, the results are accessible to the non specialist where microscopic retinal abnormalities are grossly and easily noticeable; on the other hand, results are reproducible and exceedingly quantitative in the hands of the specialist. However, as in any other imaging technique in ophthalmology, some artifacts are expected to occur. Understanding of the basic principles of image acquisition and data processing as well as recognition of OCT limitations are crucial issues to using this equipment with cleverness. Herein, we took a brief look in the past of OCT and have explained the key basic physical principles of this imaging technology. In addition, each of the several steps encompassing a third generation OCT evaluation of retinal tissues has been addressed in details. A comprehensive explanation about next generation OCT systems has also been provided and, to conclude, we have commented on the future directions of this exceptional technique. PMID- 16716640 TI - The lumbar multifidus: does the evidence support clinical beliefs? AB - The contribution of the trunk muscles to spinal stability is well established. There is convincing evidence for the role of multifidus in spinal stability. Recently, emphasis has shifted to the deep fibres of this muscle (DM) and five key clinical beliefs have arisen: (i) that DM stabilizes the lumbar spine whereas the superficial fibres of lumbar multifidus (SM) and the erector spinae (ES) extend and/or rotate the lumbar spine, (ii) that DM has a greater percentage of type I (slow twitch) muscle fibres than SM and ES, (iii) that DM is tonically active during movements of the trunk and gait, whereas SM and ES are phasically active, (iv) that DM and the transversus abdominis (TrA) co-contract during function, and (v) that changes in the lumbar paraspinal muscles associated with LBP affect DM more than SM or ES. This paper reviews the biomechanical, electromyographic, histochemical and morphological data that underpin these beliefs. Although there is support for the importance of the lumbar multifidus and the specific contribution of this muscle to intervertebral control, several of the clinical beliefs have little or no support and require further evaluation. These findings have implications for clinical practice. PMID- 16716641 TI - Associated sagittal spinal movements in performance of head pro- and retraction in healthy women: a kinematic analysis. AB - Sagittal head excursions are frequently used as diagnostic and treatment tools by physiotherapists. Retractions are performed to promote good head-on-body orientation. This study examined the regional contribution of spinal movements to head pro- and retraction in addition to the effect of a more or less restrained sitting position in healthy women. Fourteen healthy women performed seven sagittal head excursions in a more or less restrained sitting position, during which time their kinematic response was measured with an optoelectronic system. Total anterior/posterior head excursion was smaller (P=0.005) in the more restrained sitting position. In both sitting positions, approximately 60% of the total anterior/posterior head excursion originated from the cervical spine, almost 30% from the cervicothoracic spine C7-T4, and approximately 10% from thoracic regions down to T12. Middle thoracic vertical displacement was smaller (p=0.005) in the more restrained sitting position. A high correlation was found between total head excursion and the cervicothoracic unit displacements in both sitting positions (r=0.79, r=0.85, respectively). In each sitting position, the craniovertebral angle, and the tragus-C7-horizontal line decreased in protraction. Movements in the thoracic region contributed to the total head excursion. Therefore, clinicians should recognize the thoracic contribution to sagittal head excursion when using pro- and retraction as a diagnostic and treatment tool. PMID- 16716642 TI - Tissue pathophysiology, neuroplasticity and motor behavioural changes in painful repetitive motion injuries. PMID- 16716643 TI - (+)Insert smooth muscle myosin heavy chain (SM-B): from single molecule to human. AB - In smooth muscle, alternative mRNA splicing of a single gene produces four myosin heavy chain (SMMHC) isoforms. Two of these isoforms differ by the presence [(+)insert] or absence [(-)insert] of a seven amino acid insert in the motor domain. This insert enhances the kinetic properties of myosin at the molecular level but its exact role at the cell and tissue levels still has to be elucidated. This review focuses on the expression and biological functions of the (+)insert isoform. Current knowledge is summarized regarding its tissue distribution in animals and humans. Studies at the molecular, cellular and tissue levels that aimed at understanding the contribution of this isoform to smooth muscle mechanical function are presented with a particular focus on velocity of shortening. In addition, the altered expression of the (+)insert isoform in diseases and models of diseases and the compensatory mechanisms that occur when the (+)insert is knocked out are discussed. The need for additional studies on the relationship of this isoform to contractile performance and how expression of this isoform is regulated are also considered. PMID- 16716644 TI - Isolation of plasmid pKM101 in the Stocker laboratory. AB - pKM101 is a mutagenesis-enhancing resistance transfer plasmid (R plasmid) that was introduced into several tester strains used in the Salmonella/microsome mutation assay (Ames test). Plasmid pKM101 has contributed substantially to the effectiveness of the Ames assay, which is used on a world-wide basis to detect mutagens and is required by many government regulatory agencies for approval to market new drugs and other chemical agents. Widely used since 1975, the Ames test is still regarded as one of the most sensitive genetic toxicity assays and a useful short-term test for predicting carcinogenicity in animals. Plasmid pKM101, which is a deletion derivative of plasmid R46 (also referred to as R-Brighton after its origin of isolation in Brighton, England), has also been used to elucidate molecular mechanisms of mutagenesis. It was isolated in the laboratory of Professor Bruce A.D. Stocker at Stanford University as part of my doctoral research with 20 R plasmids. Professor Stocker's phenomenal insight into the genetics of Salmonella typhimurium and plasmid behavior was a major factor that led to the isolation of pKM101. This paper includes a tribute to Bruce Stocker, together with a summary of my research with mutagenesis-enhancing R plasmids and a brief discussion of the molecular mechanisms involved in pKM101 plasmid mediated bacterial mutagenesis. PMID- 16716645 TI - The equilibrium structures, vibrational spectra, NLO and directional properties of transition dipole moments of diguanidinium arsenate monohydrate and diguanidinium phosphate monohydrate. The theoretical DFT calculations. AB - For diguanidinium arsenate monohydrate and diguanidinium phosphate monohydrate the energies were minimized and the theoretical vibrational frequencies and potential energy distribution (PED) were calculated by density functional method. The 6-31++G(d,p) basis set was used. The assignment of the bands has been made on the basis of the calculated PED. For calculated equilibrium geometries two methods for determination of direction of transition dipole moments (TDM) were used. The oriented gas model was used for calculation of "static" TDM and from the other side the "dynamic" approximation of TDM by analysis of changes in internal coordination during characteristic vibrations was performed. The restricted Hartree-Fock (RHF) methods were used for calculation of the hyperpolarizability for both investigated molecules. The theoretical results are compared with experimental value of beta. PMID- 16716646 TI - Density functional theory and Hartree-Fock studies: geometry, vibrational frequencies and electronic properties of Anderson-type heteropolyanion, [XM6O24]n (X=TeVI, IVII and M=Mo, W) and [SbVW6O24]7-. AB - The geometry, IR and Raman spectra and electronic properties of Anderson-type heteropolyanions with main-group high oxidation state heteroatom, [Te(VI)Mo(6)O(24)](6-), [I(VII)Mo(6)O(24)](5-), [Sb(V)W(6)O(24)](7-), [Te(VI)W(6)O(24)](6-) and [I(VII)W(6)O(24)](5-) have been investigated using Hartree-Fock (HF) and density functional theory (DFT) methods. HF method has good results in geometry parameters but poorer than DFT method in the results of vibrational frequencies. Also we have investigated the effect of LanL2DZ augmented in the vibrational frequencies. With attention to relative charge and size of the cavity occupied by XO(6) subunit in these anions, we suggest that the general formula [XO(6)(n-)@M(6)O(18)] to describe electronic properties of these anions. PMID- 16716647 TI - Structural, spectral, linear and nonlinear optical properties of new nonlinear optical L-histidinium trichloroacetate crystals. AB - Single crystals of a new histidinium salt: L-histidinium trichloroacetate {abbreviated as LHTCA; [(C(3)N(2)H(4))CH(2)CH(NH(3))(CO(2))](+)CCl(3)COO(-)} were grown by slow evaporation of an aqueous solution at room temperature. The compound crystallizes in a non-centrosymmetric space group P2(1) of monoclinic system with cell parameters a=5.4505(18) A, b=25.769(8) A, c=9.210(2) A and beta=99.98(2) degrees. The vibrational structure of the compound confirms the presence of various functional groups in the molecule. The UV-vis-NIR spectrum shows a good transparency in the whole of the region from ultraviolet to near IR. The Kurtz powder SHG measurement confirms the frequency doubling of the crystal. Thermal behaviour of the crystals has been investigated by DSC analysis. These preliminary results suggest that LHTCA crystal can act as a potential and promising candidate for frequency doubling applications even from the near UV region to the near IR. PMID- 16716648 TI - Spectroscopy property of Ag nanoparticles. AB - Three silver nanoparticles of different size characterized by the UV-vis absorbance spectra and TEM images were prepared by citrate reduction and laser ablation with excitation of 532 and 248 nm. It is proved that all of them are effective SERS-active substrates, whereas, enhancement effect of silver colloids has not too much relation with the size and shape of the Ag nanoparticles. However, different photoluminescence spectra were observed from these three particles, indicating that the photoluminescence property of silver nanoparticles is dependent on the size. The spectra shift to higher energies with decreasing particle size. In addition, we also tentatively give the assignment of the luminescence bands. PMID- 16716650 TI - Ab initio and DFT studies of the vibrational spectra of benzofuran and some of its derivatives. AB - The vibrational spectra of benzofuran and some of its derivatives have been systematically investigated by ab initio and density functional B3LYP methods. The harmonic vibrational wavenumbers and intensity of vibrational bands were calculated at ab initio and DFT levels invoking different basis sets up to 6 311++g**. Vibrational assignments have been made and it has been found that the calculated DFT frequencies agree well in most cases with the observed frequencies for each molecule. Conformational studies have also been carried out and it is evident from ab initio calculations that 2(3H) benzofuranone is more stable than 3(2H) benzofuranone in support to our earlier semiempirical results. PMID- 16716649 TI - Charge transfer complex studies between some non-steroidal anti-inflammatory drugs and pi-electron acceptors. AB - Charge transfer (CT) complexes of some non-steroidal anti-inflammatory drugs, naproxen and etodolac which are electron donors with some pi-acceptors, such as tetracyanoethylene (TCNE), 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ), p chloranil (p-CHL), have been investigated spectrophotometrically in chloroform at 21 degrees C. The coloured products are measured spectrophotometrically at different wavelength depending on the electronic transition between donors and acceptors. Beer's law is obeyed and colours were produced in non-aqueous media. All complexes were stable at least 2 h except for etodolac with DDQ stable for 5 min. The equilibrium constants of the CT complexes were determined by the Benesi Hildebrand equation. The thermodynamic parameters DeltaH, DeltaS, DeltaG degrees were calculated by Van't Hoff equation. Stochiometries of the complexes formed between donors and acceptors were defined by the Job's method of the continuous variation and found in 1:1 complexation with donor and acceptor at the maximum absorption bands in all cases. PMID- 16716651 TI - Spectrophotometric and spectrofluorimetric methods for analysis of tramadol, acebutolol and dothiepin in pharmaceutical preparations. AB - Sensitive spectrophotometric and spectrofluorimetric methods are described for the determination of tramadol, acebutolol and dothiepin (dosulepin) hydrochlorides. The two methods are based on the condensation of the cited drugs with the mixed anhydrides of malonic and acetic acids at 60 degrees C for 25-40 min. The coloured condensation products are suitable for the spectrophotometric and spectrofluorimetric determination at 329-333 and 431-434 nm (excitation at 389 nm), respectively. For the spectrophotometric method, Beer's law was obeyed from 0.5 to 2.5 microg ml(-1) for tramadol, dothiepin and 5-25 microg ml(-1) for acebutolol. Using the spectrofluorimetric method linearity ranged from 0.25 to 1.25 microg ml(-1) for tramadol, dothiepin and 1-5 microg ml(-1) for acebutolol. Mean percentage recoveries for the spectrophotometric method were 99.68+/-1.00, 99.95+/-1.11 and 99.72+/-1.01 for tramadol, acebutolol and dothiepin, respectively and for the spectrofluorimetric method, recoveries were 99.5+/ 0.844, 100.32+/-0.969 and 99.82+/-1.15 for the three drugs, respectively. The optimum experimental parameters for the reaction has been studied. The validity of the described procedures was assessed. Statistical analysis of the results has been carried out revealing high accuracy and good precision. The proposed methods were successfully applied for the determination of the selected drugs in their pharmaceutical preparations with good recoveries. The procedures were accurate, simple and suitable for quality control application. PMID- 16716652 TI - FTIR, FT-Raman spectra and ab initio DFT vibrational analysis of 2,4-dichloro-6 nitrophenol. AB - The FTIR and FT-Raman spectra of 2,4-dichloro-6-nitrophenol (2,4-DC6NP) has been recorded in the region 4000-400 cm(-1) and 3500-100 cm(-1), respectively. The optimized geometry, frequency and intensity of the vibrational bands of (2,4 DC6NP) were obtained by the ab initio and DFT levels of theory with complete relaxation in the potential energy surface using 6-31G(d,p) and 6-311+G(d,p) basis sets. The harmonic vibrational frequencies were calculated and the scaled values have been compared with experimental FTIR and FT-Raman spectra. The observed and the calculated frequencies are found to be in good agreement. The experimental spectra also coincide satisfactorily with those of theoretically constructed bar type spectrograms. PMID- 16716653 TI - Synthesis, molecular structure and vibrational spectra of a dimeric complex formed by cobalt and glycine. AB - The structure of [Co(gly)(2)(OH)(2)].1.5(H(2)O) was solved by X-ray diffraction. It crystallizes in the space group P-1, with two independent dimmers in the unit cell. The results for the calculated vibrational spectra are in good agreement with the experimental one. The infrared spectrum and ab initio calculations are consistent with the crystallographic results. PMID- 16716654 TI - Anti-Stokes Yb3+ emission--valuable structure information in spectra of rare earth compounds measured with FT-Raman spectrometers. AB - Raman spectroscopy is a powerful and simple method which proved to be very useful in studies of solids. The most widely used Raman spectrometers are FT-Raman instruments with YAG:Nd(3+) laser as an excitation source. However, in the case of samples containing rare earth elements, the quality of FT-Raman spectra is often low due to strong fluorescence effects. We show that, in such cases, anti Stokes part of the Raman spectra often contains strong, well resolved bands identified as multiphonon-assisted emission bands of Yb(3+) present as an impurity. We show on several examples that analysis of these bands may provide useful structure information, similar to that obtained by "Eu structure probe" method in optical spectroscopy. The Yb(3+) emission can be also measured using standard luminescence detection systems. However, the application of FT-Raman system allows one to obtain good quality spectra in a much cheaper, easier and faster way (in times as short as a few seconds). Moreover, high-sensitivity of FT Raman spectrometers allows to detect even very small amounts of Yb(3+) impurity. PMID- 16716656 TI - Simple and sensitive method for spectrofluorimetric determination of dodecyl benzene sulfonic acid sodium. AB - The interaction of poly(diallyldimethyl ammonium chloride) (PDDA) with dodecyl benzene sulfonic acid sodium (DBS) has been studied by fluorescence spectra. The fluorescence of DBS can be greatly enhanced by addition of PDDA, owing to the interaction between PDDA and DBS. The enhancement intensity of fluorescence was proportional to the concentration of DBS over the range 2.5x10(-7) to 9.6x10( 5)molL(-1). Its detection limit is 3.5x10(-7)molL(-1). The method has high sensitivity and selectivity and was applied to the determination of trace amounts of DBS in water samples with satisfactory result. PMID- 16716655 TI - Vibrational spectra and normal coordinate analysis of diazepam, phenytoin and phenobarbitone. AB - Vibrational spectroscopy is an important tool for the structural investigation of the organic molecules. In the present investigation, a normal coordinate analysis has been carried out on some anti-epileptic drugs, viz. diazepam, phenytoin and phenobarbitone. Diazepam is a derivative of benzodiazepine, phenytoin is a derivative of hydanation and pheonobarbitone is a barbiturate. The infrared spectra of the compounds are recorded in the region 4000-400 cm(-1) and Raman spectra are recorded in the region 3500-50 cm(-1). From the structural point of view, diazepam, phenytoin and phenobarbitone have been assumed to C(s) point group. A systematic set of symmetry coordinates has been constructed for these compounds and Wilson's FG matrix method has been applied for the normal coordinate analysis using general quadratic valance force field. The potential energy distribution is also calculated to check the vibrational band assignments. PMID- 16716657 TI - FTIR features of lithium-iron phosphates as electrode materials for rechargeable lithium batteries. AB - The essential structural features of lithium-metal phosphates (LMP) have been studied using FTIR spectroscopy which is a sensitive tool to probe the local environment in the solid materials. Various LMP materials where M is iron have been investigated including phospho-olivine LiFePO(4), diphosphate LiFeP(2)O(7), Nasicon-type phosphate Li(3)Fe(2)(PO(4))(3) and dihydrate FePO(4).2H(2)O. Vitreous and amorphous materials are also considered. Analysis of internal and external modes of vibration allows to distinguish between the different phases and the type of cationic environment in the framework. Results corroborate the contribution of the main factors which are responsible for the complexity of the spectra, i.e. departure from ideal symmetry, interactions between polyhedra, bridging atoms and lattice distortion. PMID- 16716658 TI - A potent angiogenic factor, vascular endothelial growth factor, improves the survival of the on-going acute hepatic failure in rats. AB - Although it has been shown that a simultaneous administration of the vascular endothelial growth factor (VEGF); a potent angiogenic factor, could improve the overall survival of chemically induced acute hepatic failure (AHF) in rats, it has not been elucidated yet whether this salvage effect can be observed in the on going AHF or not. For future clinical application, we examined the effect of VEGF on the on-going AHF. A combination of d-galactosamine (Gal-N) and lipopolysaccharide (LPS) was administered to induce AHF in rats. The survival rate and several indices were compared with or without VEGF treatment at 12 and 24h after the intoxication. Even after the establishment of severe liver injury, the overall survival and the serum ALT elevation were significantly improved by treatment with VEGF. The proliferation of the hepatocytes and sinusoidal endothelial cells (SEC) was also stimulated by VEGF. Furthermore, VEGF prevented the destruction of the architecture of the hepatic sinusoids. Since VEGF significantly improved the survival of the on-going AHF, the exogenous VEGF administration may represent a feasible new therapeutic strategy for AHF in the future. PMID- 16716659 TI - Ventricular pre-excitation and cardiac hypertrophy mimicking hypertrophic cardiomyopathy in a Turkish family with a novel PRKAG2 mutation. AB - BACKGROUND: Mutations in PRKAG2, the gene for the gamma2 regulatory subunit of AMP-activated protein kinase, cause cardiac hypertrophy and electrophysiological abnormalities. We identified a novel mutation in PRKAG2 causing familial ventricular pre-excitation and severe cardiac hypertrophy. METHODS AND RESULTS: We studied 30 members of one family and 120 healthy controls. Molecular analysis of PRKAG2 gene revealed one missense mutation in exon 14 which was confirmed by restriction enzyme digestion. We identified a G to A transition, resulting in a Glu506Lys substitution in the PRKAG2 gene in 8 of the family members, who all had cardiac hypertrophy and ventricular pre-excitation. High incidence of right ventricular hypertrophy and left ventricular outflow tract obstruction are other prominent features of this novel PRKAG2 mutation. Family members without mutation had no cardiac disease. The 120 unrelated healthy individuals did not show this mutation. CONCLUSIONS: Coexistence of unexplained ventricular hypertrophy and pre excitation should prompt the diagnosis of PRKAG2 mutations and these patients should be referred for genetic analysis. The possible alteration of AMP-activated protein kinase activity due to genetic defects in PRKAG2 may serve as a template for developing more specific therapies in the treatment of patients with this mutation. PMID- 16716660 TI - Correlation between serial measurements of N-terminal pro brain natriuretic peptide and ambulatory cardiac filling pressures in outpatients with chronic heart failure. AB - BACKGROUND: Serial measurements of N-terminal pro brain natriuretic peptide (NT proBNP) have been suggested for the management of outpatients with chronic heart failure (CHF). The relationship between NT-proBNP plasma levels and central haemodynamic parameters in this setting is not known. METHODS: In 19 outpatients with CHF, NT-proBNP was related to central haemodynamic information, continuously measured with an implanted haemodynamic monitor (IHM) during 24 h of daily living activities ("24 h") and during supine rest ("rest"). In 13 patients, three to seven serial measurements were obtained with a mean time interval of 39 days (range 19-113). RESULTS: At the first visit (n=19), NT-proBNP plasma levels were dispersed over a wide range of filling pressures and not correlated with the 24 h median of the right ventricular systolic pressure (RVSP) and the estimated pulmonary artery pressure (ePAD). However, in the individual patient, serial measurements yielded significant positive correlations between NT-proBNP and RVSP (p=0.006) and ePAD (p=0.001). During "24 h" compared with "rest", the median RVSP and ePAD were elevated by 20+/-16% and 32+/-18%, respectively, and corresponded better with NT-proBNP (p<0.05). CONCLUSION: In outpatients with CHF, single measurements of NT-proBNP are not correlated with cardiac filling pressures. However, serial measurements of NT-proBNP in each individual patient show a significant positive correlation with central haemodynamic parameters and reflect changes in the haemodynamic state over time. PMID- 16716661 TI - Type II diabetic mice exhibit contractile dysfunction but maintain cardiac output by favourable loading conditions. AB - BACKGROUND: Cardiomyopathy in type II diabetes is incompletely understood. The leptin receptor-deficient (db/db) mouse is a well-accepted model of type II diabetes. To date, left ventricular contractility has not been studied in animal models of type II diabetes with in vivo load-independent parameters. AIM: To determine cardiac function in db/db mice in vivo. METHODS: Cardiac function in 12 and 24-week-old db/db and wild-type mice was assessed using a microtip-pressure conductance catheter. RESULTS: Left ventricular contractile dysfunction, measured by load-independent parameters (preload recruitable stroke work, end-systolic elastance, dP/dt-V(ed)), is present in diabetic mice from age 24 weeks onwards. Despite this contractile dysfunction, the conventional parameters cardiac output, ejection fraction and dP/dt(max) were maintained, which was due to an increased preload and decreased afterload. Ventriculo-arterial coupling was increased and mechanical efficiency significantly reduced in db/db mice. CONCLUSION: Our results demonstrate that, despite impaired cardiac contractility and mechanical efficiency, cardiac output is maintained in db/db mice by favourable loading conditions and that in vivo load-independent measurements are necessary to fully characterize cardiac performance in animal models of pathophysiological states. PMID- 16716662 TI - The effect of visors on head and facial injury in National Hockey League players. AB - There has been an increase in the number of concussions sustained by players in the National Hockey League (NHL). While wearing a helmet is now required by the NHL, the face visor remains optional. It is unknown to what degree face visors influence concussion, other head injury and eye-injury rates at the professional level. Data from the 2001-2002 NHL season were examined. It was found that wearing a face visor did not significantly influence the prevalence of concussion. Visor protection did, however, minimise eye-injuries and other, non concussion head injuries. These data suggest that, while a visor may prevent some head and eye-injuries, other measures may be necessary to reduce the number of concussions. PMID- 16716663 TI - Proficiency in coronary angiography: local experience and college requirements. AB - BACKGROUND: Proficiency in coronary angiography is a necessary skill for all cardiology trainees and is a requirement of training for the Royal Australasian College of Physicians. AIMS: The purpose of this study was to compare the local experience of advanced trainees with the College guidelines. METHODS: A retrospective analysis of fluoroscopy time and radiation exposure of the first 150 cases of three trainees between 1997 and 2001. These data were also compared to those from three experienced cardiologists. RESULTS: Advanced trainees are significantly slower than senior colleagues when starting (p<0.01) and improvements are made after 150 cases. Not all trainees improved by the same degree over the study period and most remained slower than consultants after 150 cases. Radiation doses were higher in general although there was significant individual variation between the trainee and the effect of time was diverse. CONCLUSIONS: The performance of 150 coronary angiograms as a primary operator should be considered an arbitrary number. While improvements are seen in any task when performed repeatedly, the question of when an individual is proficient is less clear. Some individualization of assessment and monitoring would seem appropriate to ensure that trainees gain proficiency in performing coronary angiography during their advanced training period. PMID- 16716751 TI - Operon prediction based on SVM. AB - The operon is a specific functional organization of genes found in bacterial genomes. Most genes within operons share common features. The support vector machine (SVM) approach is here used to predict operons at the genomic level. Four features were chosen as SVM input vectors: the intergenic distances, the number of common pathways, the number of conserved gene pairs and the mutual information of phylogenetic profiles. The analysis reveals that these common properties are indeed characteristic of the genes within operons and are different from that of non-operonic genes. Jackknife testing indicates that these input feature vectors, employed with RBF kernel SVM, achieve high accuracy. To validate the method, Escherichia coli K12 and Bacillus subtilis were taken as benchmark genomes of known operon structure, and the prediction results in both show that the SVM can detect operon genes in target genomes efficiently and offers a satisfactory balance between sensitivity and specificity. PMID- 16716752 TI - The potential clinical role of ultrasonic strain and strain rate imaging in diagnosing acute rejection after heart transplantation. AB - BACKGROUND: There has been a continued search for a more sensitive noninvasive technique for detecting sub-clinical acute rejection in heart transplant recipients. Ultrasonic deformation imaging (strain/strain rate) is sensitive in detecting sub-clinical abnormalities in regional systolic function and could potentially be sufficiently sensitive to detect changes in deformation induced by graft rejection. AIM: To assess the use of strain (S) and strain rate (SR) imaging as a noninvasive method for monitoring and diagnosing acute rejection in heart transplant recipients. METHODS AND RESULTS: A prospective preliminary study was carried out involving 31 consecutive heart transplant patients who underwent a total of 106 routine follow up endomyocardial biopsy with correlative cardiac ultrasound data. To assess regional longitudinal deformation, ultrasonic S and SR data were acquired from the intraventricular septum, left ventricular (LV) lateral and right ventricular free walls (RVFW). For radial deformation, data were obtained from the LV posterior wall (LVPW). According to the International Society of Heart and Lung Transplantation criteria, 88 biopsies (Group 1) had grade 0 or IA rejection, and 18 biopsies (Group 2) had > or =grade IB rejection. Longitudinal peak systolic S and SR were decreased (p<0.05) in Group 2, compared to Group 1 in the RVFW basal and apical segments and the basal and mid segments of the LV lateral wall. Radial peak systolic S and SR were significantly lower (p<0.001) in Group 2, compared to Group 1. CONCLUSIONS: S/SR imaging might be a good technique and an additional tool for detecting > or =IB grade of acute rejection. The myocardial deformation, as assessed by S/SR imaging could be of clinical value in monitoring and diagnosing acute rejection in heart transplant recipients and could improve patients' management by reducing the number of biopsies performed. PMID- 16716753 TI - Mobile right heart thrombus and massive pulmonary embolism. AB - The current report describes a patient with pulmonary embolism, treated unsuccessfully with heparin. Transthoracic echocardiography revealed free floating right heart thrombus. Migrating deep vein thrombus to the right heart was suspected. Transesophageal echocardiography confirmed origin of the thrombus in the inferior cava vein. Mortality rate of mobile right heart thrombus is over 40%, therefore urgent surgical embolectomy was performed with relief of symptoms. PMID- 16716754 TI - Comparison of neuropsychological outcomes after selective amygdalohippocampectomy versus anterior temporal lobectomy. AB - Transsylvian selective amygdalohippocampectomy (TSA) is an operative technique designed to spare unaffected brain tissue during surgical treatment for mesial temporal lobe epilepsy. In contrast to standard anterior temporal lobectomy (ATL), the advantages of TSA with respect to postoperative cognitive outcome are equivocal. We compared cognitive function before and after surgery in 49 patients with unilateral mesial temporal lobe seizures who underwent either ATL (n=17) or TSA (n=32). All patients received neuropsychological testing before and 1 year after surgery. The intelligence quotient (IQ) increased postoperatively in both surgical groups. Memory evaluation in the ATL group revealed a postoperative decline in nonverbal memory after right-sided resection and a postoperative decline in verbal memory after left-sided resection. In the TSA group, there was a slight postoperative decline only in verbal memory after left-sided resection, but other memory function was well preserved. In particular, there was significant postoperative improvement in verbal memory after right-sided resection. Overall, memory function was better preserved in the TSA group than in the ATL group. PMID- 16716755 TI - Sildenafil induces angiogenic response in human coronary arteriolar endothelial cells through the expression of thioredoxin, hemeoxygenase and vascular endothelial growth factor. AB - This study was undertaken to investigate the effect of phosphodiesterase-5 (PDE5) inhibitor, sildenafil, on angiogenic response in human coronary arteriolar endothelial cells (HCAEC). The cells exposed to sildenafil (1-20 microM) demonstrated significantly accelerated tubular morphogenesis with the induction of thioredoxin-1 (Trx-1), hemeoxygenase-1 (HO-1) and VEGF. Sildenafil induced VEGF and angiopoietin specific receptors such as KDR, Tie-1 and Tie-2. This angiogenic response was repressed by tinprotoporphyrin IX (SnPP), an inhibitor of HO-1 enzyme activity. Sildenafil below 1 muM has no angiogenic effect as evidenced by reduced tuborogenesis. Sildenafil along with SnPP inhibited both VEGF and Angiopoietin-1 (Ang-1) protein expression. Therefore our results demonstrated for the first time that sildenafil is a very potent pro-angiogenic factor. PMID- 16716756 TI - The complex role of PPARgamma in renal dysfunction in obesity: managing a Janus faced receptor. AB - Obesity is frequently accompanied by insulin resistance, type II diabetes, hypertension and atherosclerosis, a cluster of pathologies that are the major components of the metabolic syndrome. Obesity is a known cause for renal dysfunction that leads to two major renal pathologies: hypertension and glomerular and tubulointerstitial injury. Peroxizome proliferator activated receptors (PPARs) are transcription factors belonging to the nuclear hormone receptor superfamily with important functions in the regulation of metabolism. The role of PPARgamma isoforms in adipogenesis and vascular inflammation associated to obesity has been vastly studied and is well recognized, albeit not completely mechanistically understood. Also, the effect of various PPARgamma agonists on blood pressure reduction in different forms of hypertension, including obesity related hypertension has been reported, but the mechanisms involved are only beginning to be studied. Even less clear is the concurrent beneficial effect of PPARgamma agonists thiazolinendiones (TZD) on blood pressure reduction in different forms of hypertension and, at the same time, in some cases, the significant water retention leading to edema and heart failure. The occurrence of both these apparently opposite effects on the renal water and sodium handling suggests a complex role of PPARgamma in the kidney that is likely related to the metabolic state. Also, PPARgamma activation leads to a reduction in mesangial cell proliferation while stimulating apoptosis. TZD treatment reduces albuminuria in obese and diabetic humans and rodent models suggesting protective effects against renal tubuloglomerular injury. The focus of this review is to present and critically discuss the recent findings on the roles of PPARgamma in the kidney in direct relation to renal function and renal injury in obesity and obesity-initiated diabetes. PMID- 16716757 TI - Covered versus uncovered wallstent for malignant extrahepatic biliary obstruction: a cohort comparative analysis. AB - BACKGROUND & AIMS: We examined the use of both covered and uncovered Wallstents in patients with malignant extrahepatic biliary obstruction to determine whether use of covered Wallstents prolonged stent patency without increasing procedure related complications. METHODS: Commercially available silicone-covered Wallstents were prospectively evaluated in 98 patients who underwent insertion from December 2002-December 2004. Those data were compared with data obtained by retrospectively studying 108 consecutive patients who underwent endoscopic insertion of uncovered Wallstents between January 2000-November 2002, and for whom accurate and complete follow-up details were available. RESULTS: The mean stent patency was similar for both groups, 148.9 days (range, 3-667 days) for covered Wallstents and 143.5 days (3-910 days) for uncovered Wallstents (P = .531). The 2 groups also did not significantly differ in terms of cumulative stent patency according to the Kaplan-Meier lifetime table. The incidence of mild pancreatitis was similar in both groups, occurring in 6 patients with covered stents (6/98, 6.1%) and 2 patients with uncovered stents (2/108, 1.9%) (P = .154). Excluding those patients who had previously undergone cholecystectomy, acute cholecystitis occurred in 5 of 88 patients with covered Wallstents (6.1%) and 1 of 100 patients with uncovered stents (1%) (P = .104). Stent migration occurred in 6 patients with covered stents (6.1%) and 0 patients with uncovered stents (P = .011). CONCLUSION: Covered Wallstents might not prolong stent patency compared with uncovered Wallstents when used in the management of malignant extrahepatic biliary obstruction. PMID- 16716758 TI - Incidence of colectomy during long-term follow-up after cyclosporine-induced remission of severe ulcerative colitis. AB - BACKGROUND & AIMS: Cyclosporine (CSA) has been shown to be effective in steroid refractory ulcerative colitis (UC) and as an alternative to glucocorticosteroids in patients with severe attacks of UC. Our aim was to investigate the long-term efficacy of CSA. METHODS: We conducted a retrospective cohort study of all patients admitted to our institution with an attack of UC treated with intravenous CSA between November 1992 and October 2004. Patients who responded to intravenous CSA were switched to Neoral for 3 months. Kaplan-Meier curves were used for survival analysis with quantitative variables compared using a 2-tailed Student t test with qualitative variables and differences compared with a chi(2) analysis. RESULTS: A total of 118 (83%) of the 142 patients had an initial response to CSA and avoided colectomy during hospitalization. Of the 118 patients, 41 (35%) [corrected] required a future colectomy. The rate of colectomy in those already on azathioprine compared with those starting azathioprine concurrently with CSA was 59% vs 31%, respectively (P < .05). Also, 88% of patients already on azathioprine and requiring colectomy underwent surgery within the first year of receiving CSA. Life-table analysis shows that although only 33% of patients require colectomy at 1 year, 88% will require colectomy at 7 years. CONCLUSIONS: CSA is an effective short- to medium-term treatment for patients with severe UC but at 7 years, 88% of patients will require a colectomy. Azathioprine-naive patients have better outcomes. PMID- 16716759 TI - A phase I trial with transgenic bacteria expressing interleukin-10 in Crohn's disease. AB - BACKGROUND & AIMS: The use of living, genetically modified bacteria is an effective approach for topical delivery of immunomodulatory proteins. This strategy circumvents systemic side effects and allows long-term treatment of chronic diseases. However, treatment of patients with a living, genetically modified bacterium raises questions about the safety for human subjects per se and the biologic containment of the transgene. METHODS: We treated Crohn's disease patients with genetically modified Lactococcus lactis (LL-Thy12) in which the thymidylate synthase gene was replaced with a synthetic sequence encoding mature human interleukin-10. Ten patients were included in a placebo-uncontrolled trial. Patients were assessed daily for the presence of potential adverse effects by direct questioning and assessment of disease activity. We evaluated the presence and kinetics of LL-Thy12 release in the stool of patients by conventional culturing and quantitative polymerase chain reaction of LL-Thy12 gene sequences. RESULTS: Treatment with LL-Thy12 was safe because only minor adverse events were present, and a decrease in disease activity was observed. Moreover, fecally recovered LL-Thy12 bacteria were dependent on thymidine for growth and interleukin-10 production, indicating that the containment strategy was effective. CONCLUSIONS: Here we show that the use of genetically modified bacteria for mucosal delivery of proteins is a feasible strategy in human beings. This novel strategy avoids systemic side effects and is biologically contained; therefore it is suitable as maintenance treatment for chronic intestinal disease. PMID- 16716760 TI - Extracorporeal shockwaves induce the expression of ATF3 and GAP-43 in rat dorsal root ganglion neurons. AB - Although extracorporeal shockwave has been applied in the treatment of various diseases, the biological basis for its analgesic effect remains unclear. Therefore, we investigated the dorsal root ganglion neurons of rats following shockwave exposure to the footpad to elucidate its effect on the peripheral nervous system. We used activating transcription factor 3 (ATF3) and growth associated phosphoprotein (GAP-43) as markers for nerve injury and axonal regeneration, respectively. The average number of neurons immunoreactive for ATF3 increased significantly in the treated rats at all experimental time points, with 78.3% of those neurons also exhibiting immunoreactivity for GAP-43. Shockwave exposure induced injury of the sensory nerve fibers within the exposed area. This phenomenon may be linked to the desensitization of the exposure area, not the cause of pain, considering clinical research with a particular absence of painful adverse effect. Subsequent active axonal regeneration may account for the reinnervation of exposed area and the amelioration of the desensitization. PMID- 16716761 TI - The short-term effect of water-pipe smoking on the baroreflex control of heart rate in normotensives. AB - The aim of this study is to analyse the acute effect of water-pipe smoking on heart rate, blood pressure and the baroreflex control of heart rate. SUBJECTS AND METHODS: Non-invasive continuous methods were used for investigating inter-beat interval (IBI; ms), systolic blood pressure (SBP; mm Hg), diastolic blood pressure (DBP; mm Hg), pulse pressure (PP; mm Hg), mean blood pressure (MBP; mm Hg); baroreflex sensitivity in ms/mm Hg (BRS) and baroreflex sensitivity in Hz/mm Hg (BRSf), were determined by spectral analysis, in 20 normotensive volunteers age of 27+/-6 years (mean+/-S.D.) who served as their own control. The measurements were taken twice. The first measurement was taken before the water pipe smoking session (this is after >12 h of smoking cessation with a complete stopping of alcohol, coffee or tea consumption). The second measurement was taken during a 5 min period immediately after that session. (In the smoking session the volunteer smokes 5 g Maassel (fruit flavoured tobacco) for a period of 45 min). RESULTS: The inter-beat interval decreased (846+/-100 to 709+/-109 ms, p=0.0003), SBP increased (110+/-13 to 123+/-12 mm Hg, p=0.004), DBP increased (67+/-11 to 81+/-11 mm Hg, p=0.0002), PP decreased (43+/-10 to 41+/-9 mm Hg, p=0.46 ns), MBP increased (82+/-10 to 95+/-11 mm Hg, p=0.0001), BRS decreased (9.16+/-4 to 5.67+/ 3 ms/mm Hg, p=0.003) and BRSf (0.013+/-0.005 to 0.011+/-0.004 Hz/mm Hg, p=0.3 ns). CONCLUSIONS: Water-pipe smoking induced a high increase in heart rate, SBP, DBP, MBP and markedly impaired BRS. All of those markers are known as risk factors in cardiovascular diseases, hence it is concluded that water-pipe smoking is not as harmless as is thought. PMID- 16716762 TI - Selective regimen shift and demographic growth increase associated with the emergence of high-fitness variants of canine parvovirus. AB - The natural evolution of Canine parvovirus (CPV) is characterized by a variety of mutations, mainly in the VP1/VP2 gene. Although positive selection has been previously reported in CPV, little is known about its overall contribution to viral adaptation in the canine population. Herein, the influences of the evolutive constraints on CPV during a period of viral adaptation into a previously uninfected population are more clearly investigated. To do this, 31 sequences of VP1/VP2 gene obtained from symptomatic domestic dogs in Brazil were used, sampled from 1980 to 2000. Marked evolutionary changes in CPV associated with a process of fine-tuning adaptation were observed. Specifically, sequences from the 1980s revealed two distinct antigenic types (i.e. 2a and 2b) cocirculating in Brazil. Moreover, analysis of the selective regimen showed that 90% of the VP2 sites were conserved (d(N)/d(S)=0). In contrast, sequences from the 1990s were composed solely of CPV-2a with 96% of VP2 sites under purifying selection (d(N)/d(S)<1) and site 297 under strong positive selection (omega=4.9). Important features regarding the demographic history of CPV in Brazil were also observed. The viral population size passed through a short period of explosive growth that declined and then stabilized into a constant rate of spread. Remarkably, the explosive growth coincided with the appearance of CPV variants that presented a unique repertoire of mutations never before seen in other worldwide strains. The analysis also showed that the estimated nucleotide substitution was similar to those commonly observed in fast evolving RNA viruses. The present results demonstrated the adaptive potential of CPV to acquire, in short interval of 10 years, key mutations in the VP1/VP 2 gene that increased viral fitness and enabled the virus to disseminate even in vaccinated dogs. PMID- 16716763 TI - Application of cell-based biosensors for the detection of bacterial elicitor flagellin. AB - Cell-based biosensors, bioelectronic portable devices containing plant living cells have been used for monitoring some physiological changes induced by pathogen-derived signal molecules called flagellin. The screen-printed electrodes have been adapted for preparation of biosensors. The proton-sensitive thick films have been printed using composite bulk modified with edition of RuO(2). Obtained disposable electrodes were made possible to measure the pH change with well sensitivity and reproducibility. Tobacco cells attached to the electrode surface, cell-based biosensor, can be used for the detection of flagellin, the virulence factor of bacterial pathogen. We culture tobacco cells on the surface of such electrotransducer for several weeks and monitor of potential of cells under flagellin stimulation. The detection of the electrochemical proton gradient across the plasma membrane serves as the analytical signal. The electrode response depended upon H(+) concentration in extracellular solution. It can be conveniently observed on the surfaces of biosensors. Suitable stability and the good response time of constructed biosensors were observed. Future development of these cell-based biosensors could draw advances in selective monitoring of microbial pathogens and other physiologically active components. Moreover, this new method is much faster compared with the traditional microbial testing. PMID- 16716764 TI - Nonlinear current-voltage relationship of the plasma membrane of single CHO cells. AB - The application of electric field pulses to Chinese Hamster Ovary (CHO) cells causes membrane electroporation (MEP). If a voltage or current ramp is applied to the cellular membrane of a single CHO cell, the membrane conductance increases nonlinearly with field strength reaching saturation. In particular, the kinetics of the induced conductance changes represents the data basis for the interpretation in terms of underlying structural changes. The current/voltage characteristic is found to be continuous, but displays occasionally a sharp increase in the conductance. The step-like increases are interpreted to reflect the formation of one (or more) larger pore(s). The analysis of current clamp data yields pores of radius (r(p)) in the range of 2.5< or =r(p)/nm< or =20; the pores of the voltage clamp data are in the range of 2.5< or =r(p)/nm< or =55. The larger pores occur predominantly during hyperpolarising and less frequently during depolarising conditions, respectively. The different kinetics of pore formation in the hyperpolarising condition, where the inward field increases, and the depolarising condition, where the inward field first decreases and then increases in the opposite direction, suggests structural asymmetry with respect to the direction of the electric membrane field. At the required higher voltage, the effect of the resting potential is negligibly small. PMID- 16716765 TI - Is the quality and cost of food affected if industrially produced trans fatty acids are removed? AB - Since the start in Italy and Denmark more than 10 years ago the application of fats and oils without trans fatty acids (TFA) has increased all over the world. Today the food industry uses enormous resources to decrease the content of TFA in existing and new products. The food industry has in most cases succeeded in making consumer accepted products without TFA-most of the quality issues have been related to the technological properties of the fat- and oil-based ingredients. The change from the traditional process of selective hydrogenation to fractionation and interesterification has caused a change in the demand of vegetable oils and process capacities in the market, but until now most of these changes has been absorbed by the market without significant changes in the cost of the raw materials. PMID- 16716766 TI - The scientific basis for TFA regulations-is it sufficient? Comments from the USA. AB - We presently have definitive evidence from multiple randomized trials that trans fatty acids (TFA) have adverse effects on blood lipids, near definitive evidence that TFA increase inflammatory markers in blood, and strong evidence from prospective epidemiologic studies that high TFA intake is associated elevated risks of coronary heart disease. Food additives should be allowed to be used only if there is reasonable certainty of no harm. Thus, the present US Food and Drug Administration position of allowing TFA in the food supply is indefensible, and large numbers of Americans are dying prematurely because of its failure to act to responsibly. PMID- 16716767 TI - Double-strand break repair in bacteriophage T4: coordination of DNA ends and effects of mutations in recombinational genes. AB - Coordination of DNA ends during double-strand break (DSB) repair was studied in crosses of bacteriophage T4 in which DSBs were induced site-specifically by SegC endonuclease in the DNA of only one of the parents. Coupling of the genetic exchanges to the left and to the right of the DSB was measured in the wild-type genetic background as well as in T4 strains bearing mutations in several recombination genes: 47, uvsX, uvsW, 59, 39 and 61. The observed quantitative correlation between the degree of coupling and position of the recombining markers in relation to the DSB point implies that the two variants of the splice/patch-coupling (SPC) pathway, the "sequential SPC" and the "SPC with fork collision", operate during DSB repair. In the 47 mutant with or without a das suppressor, coupling of the exchanges was greatly reduced, indicating a crucial role of the 47/46 complex in coupling of the genetic exchanges on the two sides of the DSB. From the observed dependence of the apparent coupling on the intracellular ratio of breakable and unbreakable chromosomes in different genetic backgrounds it is inferred that linking of the DNA ends by 47/46 protein is the mechanism that accounts for their concerted action during DSB repair. A mechanism of replicative resolution of D-loop intermediate (RR pathway) is suggested to explain the phenomenology of DSB repair in DNA arrest and uvsW mutants. A "left" "right" bias in the recombinogenic action of two DNA ends of the broken chromosome was observed which was particularly prominent in the 59 (41-helicase loader) and 39 (topoisomerase) mutants. Phage topoisomerase II (gp39-52-60) is indispensable for growth in the DNA arrest mutants: the doubles 47(-)39(-), uvsX 39(-) and 59(-)39(-) are lethal. PMID- 16716768 TI - Purification of VP3 protein of infectious bursal disease virus using nickel ion immobilized regenerated cellulose-based membranes. AB - In this study, hexa-histidine tagged VP3 protein of infectious bursal disease virus (IBDV) was purified using immobilized metal ion affinity technique from the fermentation of Escherichia coli BL21 (DE3) containing a recombinant plasmid with a VP3 gene. The purification efficiencies of VP3 protein (TVP3 and DeltaTVP3) using Ni(2+)-NTA commercial agarose gels and Ni(2+)-IDA regenerated cellulose based membranes at 4 degrees C were compared. A good washing condition for removing most impurity proteins was found as 20 mM NaH(2)PO(4), 500 mM NaCl, 40 mM imidazole, pH 7.8, whereas an efficient elution condition was 20 mM NaH(2)PO(4), 500 mM NaCl, 500 or 750 mM imidazole, pH 7.8. By applying these conditions to the flow experiments, similar recovery (86-88%) and purity (98-99%) for VP3 were obtained in both gel column (1 ml gel) and membrane cartridge (four membrane disks) under the flow rate of 1.7 ml/min for protein loading and 2.7 ml/min for protein elution. Regarding that the membrane process exhibited some advantages such as shorter residence time and lower cost, a better process efficiency in a large-scale system could be expected for the Ni(2+)-IDA membranes. PMID- 16716769 TI - Sample enrichment techniques in capillary electrophoresis: focus on peptides and proteins. AB - Compared to chromatography-based techniques, the concentration limits of detection (CLOD) associated with capillary electrophoresis are worse, and these have largely precluded their use in many practical applications. To overcome this limitation, researchers from various disciplines have exerted tremendous efforts toward developing strategies for increasing the concentration sensitivities of capillary electrophoresis (CE) systems, via the so-called sample enrichment techniques. This review highlights selected developments and advances in this area as applied to the analyses of proteins and peptides in the last 5 years. PMID- 16716770 TI - Liquid chromatography method for quantifying D-threo-1-phenyl-2-palmitoylamino-3 morpholino-1-propanol (D-threo-PPMP) in mouse plasma and liver. AB - A high-performance liquid chromatography (HPLC) method was developed to measure levels of d-threo-1-phenyl-2-palmitoylamino-3-morpholino-1-propanol (d-threo PPMP) in mouse plasma and liver. d-threo-PPMP was measured by HPLC with a Luna Pheny-Hexyl column (5 microm, 250 mm x 4.6 mm) employing UV detection at 210 nm using a mobile phase of potassium phosphate buffer (20mM, pH 3.0)-acetonitrile in a 45:55 (v/v) ratio. d-threo-1-phenyl-2-pentadecanoylamino-3-morpholino-1 propanol (PC15MP) was employed as an internal standard (IS). The lower limit of quantitation (LLOQ) was 0.3 microg/ml. The assay was linear over a concentration range of 0.3-10 microg/ml, with acceptable precision and accuracy. Assayed in plasma, the intra- and inter-day validation for all coefficients of variation (R.S.D.%) were found less than 15%. The method was applied to samples from athymic (nu/nu) mice treated with d-threo-PPMP by intraperitoneal injection. d threo-PPMP levels of approximately 10-20 microg/ml ( approximately 20-40 microM) in plasma and approximately 45 microg/g in liver were obtained. The present method can be used to quantify d-threo-PPMP in mice for bioavailability and dose response studies. PMID- 16716771 TI - Hair to document exposure to glibenclamide. AB - Among the drugs that are used to incapacitate victims such as kids or elderly for sedation or for criminal gain such as sexual offences or robberies, glibenclamide, an antidiabetic was never mentioned. To document the interest of hair testing in such forensic situations, we have developed an original method to test for glibenclamide. A 30-year-old man was admitted to the Emergency Unit for coma and seizures after a party with some members of his family. Blood glucose was 0.40 g/l. A hair specimen was collected several weeks after the event and divided into two segments of 2 cm. Twenty milligrams of each segment cut into small pieces were incubated overnight in a phosphate buffer (pH 5.5), in presence of gliclazide used as internal standard (IS). A liquid/liquid extraction was realized with a mixture of diethyl ether/methylene chloride, and hair extract was separated on a XTerra MS C18 column using a gradient of acetonitrile and formate buffer. Detection of glibenclamide was achieved using two transitions: m/z 493.9 to 168.9 and 493.9 to 368.8. Linearity was observed from 5 to 1000 pg/mg (r2 = 0.956) with a limit of quantification at 5 pg/mg and a clean-up recovery of about 61%. Within-batch precision and bias were 9.0 and 9.5%, respectively. Ion suppression tested on drug-free hair was about 50%. Glibenclamide tested positive in the two consecutive segments (root to 2 cm: 23 pg/mg and 2-4 cm: 31 pg/mg). These findings were in accordance with a repetitive exposure to the drug. The concentrations were compared with those obtained after a single and a daily dose administration. In the hair of a subject receiving a single 5mg dose and collected 4 weeks later, glibenclamide was detected in the proximal segment at 5 pg/mg. After a 20 mg/day dose, the hair concentration of a subject under glibenclamide therapy was 650 pg/mg. PMID- 16716772 TI - Determination of a prostaglandin D2 antagonist and its acyl glucuronide metabolite in human plasma by high performance liquid chromatography with tandem mass spectrometric detection--a lack of MS/MS selectivity between a glucuronide conjugate and a phase I metabolite. AB - A method for the determination of a prostaglandin D(2) receptor antagonist (I, a compound being evaluated for the prevention of niacin induced flushing) and its acyl glucuronide metabolite (II) in human plasma is presented. The method utilized high performance liquid chromatography (HPLC) with tandem mass spectrometric (MS/MS) detection using an atmospheric pressure chemical ionization (APCI) interface operated in the positive ionization mode. The product ion was a radical cation generated via a homolytic bond cleavage. A chemical analog of the drug was used as internal standard (III). The acyl glucuronide metabolite (II) was detected using the same precursor-to-product ion transition used for the parent compound after chromatographic separation of I and II. Drug and metabolite were extracted using semi-automated, 96-well format solid phase extraction (SPE), and chromatography was performed using a reverse phase analytical column with an isocratic mobile phase. The chromatographic retention factor (k') of II was found to be highly sensitive to mobile phase formic acid concentration. An adjustment in mobile phase formic acid concentration improved the chromatographic separation between II and a mono-hydroxylated metabolite after an unexpected lack of MS/MS selectivity between the two molecules was observed. The dependence of retention factor on formic acid concentration (k' increased as formic acid concentration decreased) was thought to indicate polar interactions between II and the stationary phase. The stability of II in spiked human plasma was determined. The rate of hydrolysis back to parent compound was relatively low (approximately 0.1 and 0.5% per hour at room temperature and 4 degrees C, respectively) indicating that significant changes in analyte concentrations did not occur during sample processing. The concentration range of the assay was 10-2500 ng/mL for both drug and glucuronide metabolite. PMID- 16716773 TI - Isolation of lipids from photosystem I complex and its characterization with high performance liquid chromatography/electrospray ionization mass spectrometry. AB - A method for simultaneous analysis of lipids extracted from photosystem I complex was developed with high performance liquid chromatography/electrospray ionization mass spectrometry. The photosystem I complex was firstly solubilized and separated using deoxycholate polyacrylamide gel electrophoresis method after ultrasonic treatment of the sample (leaves of pea, Pisum sativum L.). The Photosystem I complexes were electrophoretically eluted from the deoxycholate polyacrylamide gel electrophoresis bands containing them, and the electron transport activity of the eluent measured as confirmation. Lipids, which were isolated from the complex having photosystem I activity, were separated and characterized with high performance liquid chromatography/electrospray ionization mass spectrometry. Five lipids, monogalactosyldiacylglycerol, digalactosyldiacylglycerol, phosphatidylglycerol, sulphoquinovosyldiacylglycerol and phosphaditylcholine were found combining with photosystem I complex. Different species of these lipids were found in the ESI mass spectra and the compositions of the acyl groups in them were determined. PMID- 16716774 TI - Sensitive quantification of chosen drugs by reversed-phase chromatography with electrochemical detection at a glassy carbon electrode. AB - Reversed-phase-high-performance liquid chromatographic method with electrochemical detection has proven to be a highly sensitive and selective method for determination of trace components in complex biological samples, and the electrochemical detector becomes an important alternative tool to ultraviolet and fluorescence detectors. A rapid and sensitive method for the accurate determination of metoclopramide, hydrochlorothiazide, imipramine and diclofenac in serum or plasma samples is described. The method is based on liquid-liquid extraction. The compounds were separated on C-18 column as stationary phase with a different binary mixture as mobile phase. Proposed method was validated with respect to specificity, linearity range, limit of detection and quantitation, precision, accuracy and successfully applied in a pharmacokinetic studies. PMID- 16716775 TI - Determination of lycopene and beta-carotene by high-performance liquid chromatography using sudan I as internal standard. AB - A new method, using sudan I as internal standard to determine the content of lycopene or beta-carotene in samples, was developed. According to UV-vis absorption spectra, sudan I, lycopene and beta-carotene all had absorption peaks at 450 nm. They could be separated absolutely by high-performance liquid chromatography (HPLC) with retention time of 2.7, 6.6 and 10.1 min, respectively. The related equations between sudan I and lycopene or beta-carotene content were obtained and verified by determining the content of lycopene or beta-carotene in Blakeslea trispora cells. The relative error was below 1.4% for determining lycopene content and below 1.9% for beta-carotene. Intra-day variability for lycopene determination was less than 3.4% and for beta-carotene was less than 1.4%. The mean recovery of lycopene or beta-carotene was 96.1 and 97.9%, respectively. PMID- 16716776 TI - LC/MS/MS analysis of quaternary ammonium drugs and herbicides in whole blood. AB - Quaternary ammonium drugs (atracurium, bretylium, edrophonium, ipratropium, mivacurium, neostigmine, pancuronium and rocuronium) and herbicides (difenzoquat, diquat and paraquat) in human whole blood were analysed by LC/MS/MS with positive electrospray ionisation (ESI), following extraction with Bond Elut LRC-CBA cartridges. Internal standards were benzyldimethylphenylammonium chloride monohydrate and ethyl viologen for drug and herbicide analysis, respectively. Ion pair chromatography used heptafluorobutyric acid (15 mM)-ammonium formate (20 mM) buffer adjusted to pH 3.30 with formic acid and a linear gradient from 5 to 90% methanol run over 18 min. Recoveries ranged from 79.7 to 105.1%, detection limits were between 3.6 and 20.4 ng/ml and the intra- and inter-day precisions were less than 18.6% at a concentration of 10 ng/ml. The method was applied to a case of accidental paraquat poisoning in which the concentration of paraquat in blood was 0.64 mg/l, which is within the range associated with fatal paraquat poisoning. PMID- 16716777 TI - Circular dichroism studies of subtilisin Carlsberg immobilised on micron sized silica particles. AB - Immobilised enzymes are widely used in industry, but the reasons for loss of activity of such biocatalysts are usually not known. We have used circular dichroism (CD) to investigate the structure of one such system, i.e., subtilisin Carlsberg (SC) immobilised on silica gel particles (60 microm). A number of technical problems have to be overcome in order to obtain appropriate data from which conclusions can be drawn. A rotating cell holder has been developed to avoid sedimentation of the silica particles during the collection of spectra. By moving the cell holder as close as possible to the detector window, the effects of differential scattering can be minimised. However, the effects of absorption flattening limit the extent to which reliable quantitative information on secondary structure content can be obtained from far UV CD studies. We have used an empirical approach based on absorbance units derived from the high-tension voltage to correct for absorption flattening effects. After applying the correction there was satisfactory agreement with the solution spectra. Comparison of the fresh and used (inactive) SC-silica gel spectra in organic media reveals substantial change in the secondary structure. Additional evidence for loss of native conformation is provided by the significant decrease in the near UV CD spectrum. These results for the first time clearly demonstrate the origin of enzyme instability in the immobilised state. PMID- 16716779 TI - The role of bright liver echo pattern on ultrasound B-mode examination in the diagnosis of liver steatosis. AB - AIM: The observation of bright liver echo pattern on ultrasound is commonly considered a sign of hepatic steatosis. However, the interference of liver fibrosis on sensitivity and specificity of bright liver echo pattern has caused many to question its effectiveness as a diagnostic tool. The objective of this study was to evaluate the sensitivity, specificity and predictive values of bright liver echo pattern for liver steatosis. PATIENTS AND METHODS: We studied 235 consecutive patients suspected of having liver disease of various aetiologies. Median age was 52 years (range, 17-72 years), and there was a male/female ratio of 1:18. All patients underwent ultrasound examination before liver biopsy and was performed by two operators. The presence or absence of bright liver echo pattern and posterior attenuation or areas with different patterns of fat infiltration were noted. Histologic evaluation was performed and graded by Ishak score. Steatosis was categorised as absent, 0-2%, 3-29% to 30-49% or >50%. RESULTS: Interobserver concordance was high. Bright liver echo pattern was found in 67% of patients with steatosis of any degree and 89% of patients with steatosis of >or=30%. Only three patients without steatosis, who had a low Ishak score, demonstrated bright liver echo pattern on ultrasonography. The sensitivity, specificity, positive predictive value and negative predictive value of bright liver echo pattern for steatosis were 64%, 97%, 96.0% and 65%, respectively. Among the subgroup of patients who had steatosis of >or=30%, the sensitivity, specificity, positive predictive value and negative predictive value of bright liver echo pattern were 91%, 93%, 89% and 94%, respectively. The sensitivity, specificity, positive predictive value and negative predictive value of posterior attenuation and/or skip areas associated with bright liver echo pattern for steatosis were 89.7%, 100%, 100% and 92.3%, respectively. Univariate analysis showed bright liver echo pattern to be associated only with steatosis and not with fibrosis. CONCLUSION: We concluded that the presence of bright liver echo pattern is a sign of liver steatosis and that liver fibrosis does not interfere with ultrasound measurements. Posterior attenuation and/or skip areas are closely related to steatosis of >or=30%. PMID- 16716778 TI - Hybrid spectrin type repeats produced by exon-skipping in dystrophin. AB - Dystrophin is the protein whose defect underlies Duchenne Muscular Dystrophy, DMD, a common (1:3500 male births) and fatal condition in which muscle tissue deteriorates leading to death in the second or third decade of life. Dystrophin is coded for by the largest human gene, and one of the most complex. It is translated from at least 7 distinct promoters, with the largest transcripts (which are the ones involved in DMD) containing 79 exons over >2.5 Mbp [K.F. O'Brien, L.M. Kunkel, Dystrophin and muscular dystrophy: past, present, and future, Mol. Genet. Metab. 74 (2001) 75-88, H.M. Sadoulet-Puccio, L.M. Kunkel, Dystrophin and its isoforms, Brain Pathol. 6 (1996) 25-35]. Exacerbating this complexity, it has recently been shown that dystrophin is subject to extensive alternative RNA processing, potentially producing a wide variety dystrophin variants [M. Sironi, R. Cagliani, U. Pozzoli, A. Bardoni, G.P. Comi, R. Giorda, N. Bresolin, The dystrophin gene is alternatively spliced throughout its coding sequence FEBS Lett 517 (2002) 163-166]. The structure of the dystrophin protein is highly modular, with the most common module being a motif termed the spectrin type repeat, or STR, of which there are 24. Each STR is roughly coded for by two exons, and the most common type of multiple exon-skipping events start and end at introns in the middle of STRs [R.G. Roberts, A.J. Coffey, M. Bobrow, D.R. Bentley, Exon structure of the human dystrophin gene Genomics 16 (1993) 536-538, M. Koenig, L.M. Kunkel, Detailed analysis of the repeat domain of dystrophin reveals four potential hinge segments that may confer flexibility, J. Biol. Chem. 265 (1990) 4560-4566]. This would produce fractional STR modules, however, the concept of STRs as proteins domains makes the viability of such fractional motifs questionable. However, certain of these events produce pairs of potentially complementary fractional domain that might reassemble into a hybrid STR motif. We have constructed model fragment corresponding to one such exon-skipping event, and show that the hybrid STR so produced is viable, and furthermore that some of the properties of the protein containing it differ substantially of the native, un-skipped parent. PMID- 16716780 TI - The derivation of clinical-grade human embryonic stem cell lines. AB - The pluripotent nature of human embryonic stem cells (hESC) has attracted great interest in using them as a source of cells or tissue in cell therapy. However, in order to be used in regenerative medicine, the pluripotent hESC lines should be established and propagated according to good manufacturing practice quality requirements. The cultures should be animal substance free in order to exclude the risk of infections and immunogenity. They should also be genetically and epigenetically normal. The detailed molecular mechanisms of their pluripotency are still not defined. Using human feeder cells, a medium containing only human proteins, the mechanical isolation of the inner cell mass and mechanical passaging of hESC, is a safe option until a functional defined medium containing physiological concentrations of regulatory factors is available. PMID- 16716781 TI - Post-genomic vaccine development. AB - For over a century, vaccines were developed according to Pasteur's principles of isolating, inactivating and injecting the causative agent of an infectious disease. The availability of a complete microbial genome sequence in 1995 marked the beginning of a genomic era that has allowed scientists to change the paradigm and approach vaccine development starting from genomic information, a process named reverse vaccinology. This can be considered as one of the most powerful examples of how genomic information can be used to develop therapeutic interventions, which were difficult or impossible to tackle with conventional approaches. As the genomic era progressed, it became apparent that multi-strain genome analysis is fundamental to the design of universal vaccines. In the post genomic era, the next challenge of the vaccine biologist will be the merging of the vaccinology with structural biology. PMID- 16716783 TI - The abnormal urinalysis. AB - The urinalysis is one of the most commonly performed tests in pediatrics, and whether expected or incidental, abnormal findings are common. Understanding the strengths and limitations of the urinalysis allows one to maximize its use as a screening tool while avoiding expensive and unnecessary evaluations. This article reviews the significance of abnormal results on urine dipstick testing and urine microscopy. Causes of false positive and false negative results are summarized. Initial diagnostic considerations of abnormal urinalysis findings are provided where appropriate. PMID- 16716784 TI - Genitourinary imaging techniques. AB - In the current era of pediatric uroradiology, use of nuclear medicine, ultrasonography, CT, and MRI has been valuable in the identification and management of genitourinary diseases. Excellent information about the renal parenchyma and renal function is currently attainable with current cross sectional imaging techniques that can identify tissue differentiation of lesions, distinguish dilatation of the pelvocalyceal system, and determine margins of the kidney and perirenal space. Invasive angiography is limited in application specifically to vascular diseases, although they are uncommon in childhood. Because of these newer techniques, intravenous urography has lost its position as the "cornerstone" of urinary tract imaging and is used mainly to identify pathologic conditions of the ureters. PMID- 16716785 TI - Urinary incontinence. AB - Urinary incontinence is a common primary care problem. With the proper understanding of normal and abnormal bladder function, pediatricians can create a protocol that will benefit most children seen with this condition. Emphasis should be placed on primary therapy: establishing healthy dietary habits, treating constipation, maintaining appropriate hygiene, and developing a regular voiding pattern. When the initial evaluation identifies an anatomic abnormality or when primary therapy fails, referral to a pediatric urologist is warranted. PMID- 16716786 TI - Pediatric urinary tract infections. AB - Pediatric urinary tract infections are common. These infections have been recognized as a source of acute morbidity and long-term medical consequences in adulthood. There are various risk factors and clinical presentations in children with urinary tract infections. The main objectives in management include prompt diagnosis, appropriate antimicrobial therapy, identification of anatomic anomalies, and, in select patients, long-term follow-up. PMID- 16716787 TI - Evaluation of gross hematuria. AB - Gross hematuria in children is a common complaint that often leads patients to seek urgent care. The diagnostic evaluation can be chosen based on specific patient history and physical examination. When a patient is asymptomatic, hypercalciuria and mild forms of glomerulonephritis are common causes of gross hematuria. Although they are less common in children, special care should be taken to investigate for renal and bladder tumors. PMID- 16716788 TI - Vesicoureteral reflux. AB - Vesicoureteral reflux, the abnormal flow of urine from the bladder into the ureter, is one of the most common congenital anomalies found in children. The association of vesicoureteral reflux with urinary tract infections and renal scarring has important clinical implications. New insights into pathogenesis and new surgical techniques are changing the approach to the management of this disorder. PMID- 16716789 TI - Prenatal hydronephrosis. AB - Hydronephrosis is the most common genitourinary tract anomaly identified on prenatal ultrasound studies. Ureteropelvic junction obstruction accounts for approximately 50% of the cases of prenatally detected hydronephrosis. Postnatal evaluation allows for the identification of the cause and further management. Rarely, in utero intervention may be performed for severe oligohydramnios associated with hydronephrosis. Prenatal consultation with a pediatric urologist is useful in decreasing parental anxiety and facilitating postnatal management. PMID- 16716790 TI - Recent advances in external genitalia. AB - In the last decade, significant progress in the knowledge and management of external genital anomalies has been achieved, including an improved understanding of epidemiology and pathophysiology of these anomalies, the identification of etiologic genetic defects, and significant improvements in surgical approaches that have decreased complications and improved cosmetic outcomes. We highlight the most clinically important advances of the commonly encountered external genital anomalies. PMID- 16716791 TI - Surgical management of urolithiasis. AB - Pediatric patients with urolithiasis present unique challenges. Interventional techniques developed for adult patients have been adopted and adapted to facilitate effective and safe treatment in this population. Management must be stratified and individualized, taking into account the many factors described in this article. Long-term follow-up and metabolic evaluation are essential components of the overall treatment strategy. Interventional management will continue to evolve with progressive refinements in instrumentation and techniques. PMID- 16716792 TI - Medical evaluation and treatment of urolithiasis. AB - Nephrolithiasis is responsible for 1 in 1000 to 1 in 7600 pediatric hospital admissions annually throughout the United States. Seventy-five percent of children with nephrolithiasis have an identifiable predisposition to stone formation. This article reviews the different causes and disease states associated with nephrolithiasis in the pediatric population. The initial evaluation and the metabolic evaluation of children with nephrolithiasis are reviewed. Treatment modalities for the different stone types are also described. PMID- 16716793 TI - Hypertension in children and adolescents. AB - Hypertension is one of the most common health problems in the United States and a powerful independent risk factor for cardiovascular and renal disease. Until recently, the incidence of persistent hypertension in children has been low, with a range of 1% to 3%. Recent data indicate that over the last decade, however, average blood pressure levels have risen substantially among American children. Obesity and other lifestyle factors, such as physical inactivity and increased intake of high-calorie, high-salt foods, are thought to be responsible for this trend. Hypertension in children is currently recognized as an important health issue. There is increasing evidence that hypertension has its antecedents during childhood, because adult blood pressure often correlates with childhood blood pressure. Hypertension in children also is viewed as a significant risk factor for the development of cardiovascular disease in adulthood. PMID- 16716794 TI - Pediatric urologic emergencies and urgencies. AB - The more common urologic problems seen in infancy and childhood that require urgent or emergent pediatric urologic referral are discussed, including a brief description of the usual presenting signs or symptoms, differential diagnoses, proper diagnostic work-up recommended before or at the time of referral, and the usual therapeutic course of management after evaluation by the urologist. These conditions include the acutely swollen scrotum, scrotal masses, penile swelling and erythema, exstrophy, hematuria, urinary retention, abdominal masses, and various genital abnormalities including interlabial masses, hypospadias with any degree of cryptorchidism, and ambiguous genitalia. This article is designed to provide the primary care practitioner with a focused review and a useful resource for managing children who have genitourinary abnormalities in the hospital or clinic setting. PMID- 16716795 TI - "What's really going on down there?" A practical approach to the adolescent who has gynecologic complaints. AB - The purpose of this article is to review the common complaints related to the vaginal area in pubertal girls and offer an approach to those presenting with nonurologic perineal problems or issues related to emerging sexuality. Included is a discussion of the symptoms, diagnosis, and treatment of physiologic discharge, vulvovaginal candidiasis, bacterial vaginosis, trichomoniasis, gonorrhea, and chlamydia. Other vulvovaginal complaints are considered, and screening tests that can help differentiate the origin of the problem as urologic or gynecologic are briefly addressed. PMID- 16716797 TI - Systemic therapy for ovarian cancer: current status and new treatments. AB - Current systemic therapy for ovarian cancer consists of a combination of carboplatin and paclitaxel. While the majority of patients achieve clinical complete remission after six cycles of chemotherapy, the relapse rate stands at over 50%. Median survival time for patients after recurrence is approximately 2 years. New treatment approaches for patients with advanced ovarian cancer include consolidation and maintenance therapy, intraperitoneal administration of cytotoxic agents, new combination chemotherapy regimens, the development of new cytotoxic agents, and molecular-targeted therapies. These agents will be evaluated either singularly or with chemotherapy. Currently, the Gynecologic Oncology Group is evaluating a combination of bevacizumab together with paclitaxel and carboplatin in previously untreated patients with advanced ovarian cancer. This trial is based on phase II data that suggest that bevacizumab as a single agent has significant activity in patients with recurrent ovarian cancer. In addition, the Gynecologic Oncology Group will be conducting phase II trials of different combinations of intraperitoneal chemotherapy in an effort to decrease toxicity associated with current intraperitoneal regimens that have shown an improvement in survival in patients with small-volume stage III disease. The Gynecologic Oncology Group will also be conducting a trial of maintenance therapy in patients who enter clinical complete remission with paclitaxel plus carboplatin, comparing observation with monthly paclitaxel or monthly paclitaxel poliglumex. Novel new cytotoxic and biologic agents are also being evaluated as single agents in phase II trials in patients with recurrent ovarian cancer. PMID- 16716799 TI - Optimizing gemcitabine regimens in ovarian cancer. AB - Ovarian cancer represents the leading cause of death from gynecologic neoplasms. The chance of response to secondary treatment is currently disappointing; few agents have shown notable activity in recurrent/progressive patients. Among these agents, gemcitabine represents one of the most interesting newer antineoplastic agents, showing significant activity, synergism with cisplatin, and a mild toxicity profile in both platinum-sensitive and platinum-resistant (and also taxane-pretreated) recurrent/progressive patients. Moreover, first-line combination chemotherapy including gemcitabine has shown promising response rates in phase I and II studies. The ongoing phase III, five-arm, randomized Gynecologic Oncology Group Protocol 182/International Collaborative Ovarian Neoplasm 5 study should clarify the clinical impact of the addition of a third drug to the standard paclitaxel plus carboplatin treatment regimen. PMID- 16716798 TI - Management of platinum-sensitive recurrent ovarian cancer. AB - The majority of patients with ovarian cancer will relapse despite state-of-the art first-line surgery and chemotherapy. There are two subgroups of patients with recurrent ovarian cancer: those with platinum-resistant disease and those with platinum-sensitive disease. Re-treatment with single-agent platinum has long been considered standard therapy for patients with platinum-sensitive disease, and, based on its favorable therapeutic profile, carboplatin has become the treatment agent of choice. High response rates are seen with platinum agents used in combination with paclitaxel or gemcitabine. The International Collaborative Group for Ovarian Neoplasia (ICON) and the Arbeitsgemeinschaft fur Gynaekologische Onkologie Studiengruppe Ovarialkarzinom (AGO-OVAR) recently conducted a trial (ICON4/AGO-OVAR-2.2) comparing platinum monotherapy with platinum plus paclitaxel combined. Results showed that overall survival and progression-free survival are improved by combination therapy. Similarly, a significant benefit in progression free survival for carboplatin plus gemcitabine versus carboplatin monotherapy was seen in the Gynecologic Cancer InterGroup trial. The toxicity profiles and schedules of carboplatin plus paclitaxel and carboplatin plus gemcitabine are different, with the taxane combination having greater neurotoxicity and alopecia, less hematologic toxicity, and requiring longer drug infusions (although fewer days of treatment per cycle) than the gemcitabine combination. Based on the results of these two trials, combination chemotherapy should be considered the standard treatment of recurrent platinum-sensitive ovarian cancer. The choice of treatment needs to take into account the increase in side effects when using combination chemotherapy compared with carboplatin monotherapy, and the different toxicities of the two combination regimens. PMID- 16716800 TI - The role of gemcitabine in first-line treatment of advanced ovarian carcinoma. AB - The combination of paclitaxel and carboplatin is now established as the standard first-line chemotherapy regimen for advanced ovarian cancer. Ways in which this standard therapy can be further improved are being investigated, and several approaches have been taken. One approach is to integrate a different cytotoxic agent into the standard combination. Gemcitabine is one such cytotoxic agent that is of particular interest because it has activity in disease resistant to treatment with paclitaxel plus carboplatin, and also has a different mechanism of action to that of the two agents in the standard combination. Gemcitabine plus cisplatin has shown activity in phase II trials and results are awaited from a phase III trial comparing paclitaxel plus carboplatin with gemcitabine plus carboplatin doublets. Using agents sequentially has been shown to be as effective as using them in combination; therefore, using a gemcitabine plus platinum doublet sequentially with paclitaxel plus carboplatin has also been studied. A phase III trial has closed and results are expected shortly. Triplet combinations have also been shown to be effective in early stage trials, although dose limiting myelosuppression occurs with gemcitabine plus paclitaxel plus carboplatin. Two phase III trials of this triplet have finished and are awaiting data maturation. Lastly, a sequential triplet of gemcitabine plus paclitaxel plus carboplatin has been investigated, but high pulmonary toxicity led to the study being halted. The role of gemcitabine in first-line advanced ovarian cancer will become much clearer once the results from the various phase III trials are published. PMID- 16716801 TI - Unresolved issues in the chemotherapeutic management of gynecologic malignancies. AB - Despite substantial documented progress in the chemotherapeutic management of women with gynecologic malignancies, a number of highly clinically relevant issues remain unresolved. In ovarian cancer, questions asked by practicing oncologists, patients, and their families include the role of maintenance chemotherapy in routine management of the disease, determining which patients with early stage disease do not require cytotoxic therapy, defining the most appropriate management paradigm for both recurrent and resistant cancer, determining which taxane to combine with carboplatin in the first-line regimen, and developing an optimal intraperitoneal primary chemotherapy regimen for small volume residual advanced disease. In endometrial cancer, important issues include defining first-line chemotherapy for metastatic or recurrent disease and mixed mesodermal endometrial cancer, and determining whether there is a role for adjuvant cytotoxic chemotherapy in the setting of high-risk early stage cancer. Finally, in cervical cancer, there is a critical need to determine the most appropriate treatment strategy for metastatic cancer, as well as to find an effective second-line treatment option for platinum-resistant disease. It is reasonable to anticipate that future clinical trials will answer these and other unresolved issues, just as past well-designed studies have resulted in major improvements in both the survival and quality of life of women with this group of malignancies. PMID- 16716803 TI - Immunomodulating capacity of commercial fish protein hydrolysate for diet supplementation. AB - Dietary proteins harbour bioactive peptides that can be released by a fermentation process. Fish proteins are a valuable and little-exploited source of potentially active biopeptides. The aim of this research was to evaluate the effects of a commercially available fermented fish protein concentrate (Seacure) (FPC) derived from a fermentation process, on the mucosal immune response in a murine model. BALB/c mice received the FPC or the non-fermented powder at different concentrations (0.20, 0.25 or 0.30mg/ml) for 2, 5 or 7 consecutive days. At the end of each feeding period, histological studies of the gut were carried out and the phagocytic activity of peritoneal macrophages, the number of IgA+ cells in the small intestine lamina propria and bronchial tissue and the number of IL-4+, IL-6+, IL-10+, IFNgamma+ and TNFalpha+ cells in the small intestine lamina propria were determined. Different accumulative doses of FPC did not induce any inflammatory immune response and the normal morphology of the small intestine was not affected. Phagocytic activity of peritoneal macrophages was enhanced following FPC administration at 0.3mg/ml for 7 consecutive days. The number of IgA+ cells increased in the small intestine lamina propria but not in the bronchial tissue. IL-4, IL-6 and IL-10 were all significantly increased in the lamina propria of the small intestine of animals that received FPC. At the same time, some pro-inflammatory cytokines such as IFNgamma and TNFalpha also increased, but the intestinal homoeostasis was maintained and no tissue damage was observed. We conclude that FPC is an immunomodulating food with a demonstrated capacity to enhance non-specific host defense mechanisms. PMID- 16716802 TI - Macrophage antioxidant enzymes regulate Chlamydia pneumoniae chronicity: evidence of the effect of redox balance on host-pathogen relationship. AB - Latency, chronicity and recurrent nature are the features of Chlamydia pneumoniae biology which play a central role in the course and outcome of C. pneumoniae-host interaction. Since redox status is directly an indicator of inflammatory response via molecular signaling mechanisms, we decided to study the regulatory role of macrophage cellular redox balance on the molecular indices of C. pneumoniae chronicity. We examined GSH-GSSG status, the activities of antioxidant enzymes (SOD, GPx and gamma-GCS), along with their protein and gene expression, the MOMP and cHSP-60 protein and gene expression, and the consequence of redox balance on the establishment of productive infection in macrophages. Results showed that C. pneumoniae caused changes in GSH-GSSG levels, antioxidant enzymes activity, mRNA gene and protein expression in macrophages. The relevance of this to the state and status of C. pneumoniae in macrophages was assessed by inhibitor induced attenuation of antioxidant enzymes and there was evidence that, while SOD attenuation did not significantly affect MOMP and cHSP-60 gene and protein expression, gamma-GCS attenuation increased cHSP-60 gene and protein expression. The increase in molecular evidence of chronic forms of C. pneumoniae (cHSP-60) was consistent with decrease in normal forms of C. pneumoniae. These findings reflect the importance of redox balance modulation on the outcome of C. pneumoniae infection in macrophages, a significant process in the pathogenesis of chlamydial diseases. PMID- 16716804 TI - Effects of intravenous immunoglobulin and methylprednisolone on human umbilical vein endothelial cells in vitro. AB - Endothelial cells (ECs) do more than just play a role in distinguishing blood and tissues. These cells are also influenced by various chemical mediators present in the blood and tissues. In addition, they produce diverse cytokines, chemokines, adhesion molecules, and growth factors. Therefore, ECs are actively involved in the inflammatory and immune response. We investigated the effects of intravenous immunoglobulin (IVIG) and methylprednisolone (MP) on activated human ECs, by examining the individual and combined effects of the drugs. Human umbilical vein ECs (HUVECs) obtained from the umbilical cords of healthy newborns were cultured. After the HUVECs were treated with interleukin (IL)-1beta, the effects of IVIG and/or MP on the activated HUVECs were assessed by cell proliferation, mRNA expression, and the production of vascular cell adhesion molecule (VCAM)-1, IL 1beta, and vascular EC growth factor (VEGF). IVIG and MP inhibited HUVEC proliferation. IVIG and MP significantly down regulated mRNA expression and the production of VCAM-1, IL-1beta, and VEGF. The combination of IVIG and MP generally showed a greater suppressive effect on mRNA expression and on the production of VCAM-1, IL-1beta, and VEGF. Our results suggest that some of the corticosteroid-sparing effects of IVIG observed in patients with severe asthma could be related to a decreased ability of ECs to proliferate, and to a down regulation of the expression of molecules involved in the onset and progression of airway inflammation. PMID- 16716805 TI - Differential inhibitory effects of baicalein and baicalin on LPS-induced cyclooxygenase-2 expression through inhibition of C/EBPbeta DNA-binding activity. AB - To evaluate the possible mechanisms responsible for the anti-inflammatory effects of baicalein or baicalin, lipopolysaccharide (LPS)-induced inflammatory responses in cultured Raw 264.7 cells were studied. In the present study, baicalein and baicalin, a flavonoid present in the root of Scutellaria baicalensis Georgi, were examined for their effects on LPS-induced cyclooxygenase-2 (COX-2) gene expression in Raw 264.7 macrophages. Baicalein, but not baicalin, inhibited COX-2 gene expression in LPS-induced Raw 264.7 cells. However, both polyphenolic compounds inhibited LPS-induced inducible nitric oxide synthase (iNOS) protein expression, iNOS mRNA expression, and NO production in a dose-dependent manner. To investigate the mechanism by which baicalein inhibits COX-2 gene expression, we examined activation of mitogen-activated protein kinases (MAPKs) in Raw 264.7 cells. We did not observe any significant change in the phosphorylation of MAPKs between baicalein- and baicalin-treated cells. Baicalein and baicalin had no effect on LPS-induced nuclear factor-kappaB (NF-kappaB) and cAMP response element binding protein (CREB) DNA binding activity. Baicalein, but not baicalin, significantly inhibited the DNA binding activity of CCAAT/enhancer binding protein beta (C/EBPbeta) These results indicated that differential effects of baicalein and baicalin on COX-2 gene expression in LPS-induced Raw 264.7 cells were mediated through inhibition of C/EBPbeta DNA binding activity. Taken together, these results suggest that baicalein acts to inhibit inflammation through inhibition of COX-2 gene expression through blockade of C/EBPbeta DNA binding activity. PMID- 16716806 TI - Se-methylselenocysteine enhances PMA-mediated CD11c expression via phospholipase D1 activation in U937 cells. AB - CD11c/CD18 is expressed primarily on myeloid cells, where its expression is regulated both during differentiation and during monocyte maturation into tissue macrophages, and is also a receptor for fibrinogen and lipopolysaccharide (LPS). We focused on the molecular mechanisms leading to the activation of CD11c expression in differentiating U937 cells. During phorbol myristate acetate (PMA) induced differentiation of U937 cells, we found that the mRNA expression of CD11c was increased. Se-methylselenocysteine (Se-MSC) potentiated up-regulation of CD11c expression and its promoter activity and increased PLD1 activity without affecting the level of PLD1 protein in PMA-treated cells. To examine the regulation mechanism of PMA and Se-MSC on CD11c gene expression through the activation of PLD1, we analyzed changes in the CD11c mRNA level and the promoter activity following treatment of a selective PLD inhibitor n-butanol. The combinatory effect of PMA and Se-MSC on CD11c gene expression was abolished by n butanol in a dose-dependent manner. Further, introduction of PLD1 gene into U937 cells increased CD11c mRNA expression and activated CD11c promoter activity in a dose-dependent manner. These results showed that Se-MSC increased PMA-induced CD11c expression through the activation of PLD1 signaling pathway. To our knowledge, this is the first report that expression of the CD11c gene is regulated by PLD1 and is enhanced by Se-MSC during PMA-induced U937 differentiation. PMID- 16716807 TI - Differences in CXCR4-mediated signaling in B cells. AB - Among all chemokine receptors CXCR4 possesses a unique response profile and distinguishes itself through a prolonged signaling capacity. Here, we investigated the signaling capacity of CXCR4 to its so far known unique ligand CXCL12 in B cell lines and primary CD19(+) B lymphocytes. During lymphopoiesis, CXCR4 is continuously expressed on the surface of B cells. However, its signaling profile changes inasmuch preB and proB cells migrate towards CXCL12, mobilize intracellular calcium and activate the small GTPases Rac1 and Cdc42, whereas mature B cells do not show these responses, albeit the cells retain the capability to migrate in response to CXCL13 and CCL21. By contrast, stimulation of B cells with CXCL12 at all stages of development results in the activation of the MAP-kinase cascade and in rapid CXCR4 internalization. The pathways leading to ERK1/2 activation are different in preB and mature B cell lines. In either case, ERK1/2 activation is pertussis toxin sensitive, but only in mature B-cells inhibition of PI3-kinase causes an almost complete block of ERK1/2 activation. Taken together, the results show that CXCR4 changes its coupling to downstream signal-transduction pathways in B cells, suggesting that receptor activity may depend on accessory proteins. PMID- 16716808 TI - Identification of a rat bone marrow-derived dendritic cell population which secretes both IL-10 and IL-12: evidence against a reciprocal relationship between IL-10 and IL-12 secretion. AB - The qualitative nature of immune responses induced by dendritic cells (DCs) is influenced by the balance of pro-inflammatory (e.g. IL-12) and anti-inflammatory (e.g. IL-10) cytokines that they secrete. Evidence to date suggests that IL-12 and IL-10 secretion is reciprocally regulated and that IL-10 inhibits IL-12 secretion. This study identifies a population of resting, immature rat bone marrow-derived DCs (BMDCs) which secretes IL-10, the IL-12(p70) heterodimer and the free IL-12(p40) subunit, the latter in vast excess of IL-12(p70). Counter intuitively, activation with LPS induces the secretion of high and equivalent levels of IL-10 and IL-12(p40), but only quantitatively small increases in IL 12(p70). Neutralization of IL-10 increased the secretion of IL-12(p40) by resting BMDCs, but decreased IL-12(p40) secretion by LPS-activated BMDCs. Pre-incubation of resting BMDCs for 24h with neutralizing antibody to IL-10 reduced the subsequent secretion of IL-10 in allogeneic cultures of Lewis CD3(+) T cells with resting and LPS-activated Wistar BMDCs, and enhanced IL-12(p40) secretion in allogeneic cultures with LPS-activated BMDCs. IL-10 neutralization had no effect on the levels of IL-12(p70), IFN-gamma or IL-4 in allogeneic cultures. In summary, this study has identified a population of rat BMDCs that secretes low levels of bioactive IL-12(p70), but high levels of IL-10 and IL-12(p40). These findings argue against the concept that there is a reciprocal relationship between IL-10 and IL-12 secretion. They might also have implications for understanding the role of DCs in post-activation qualitative skewing of immune responses. PMID- 16716811 TI - Pathology of the pleura. AB - The pleura and lung are intimately associated and share many pathologic conditions. Nevertheless, they represent two separate organs of different embryonic derivation and with different yet often symbiotic functions. In this article, the authors explore the pathologic manifestations of the many conditions that primarily or secondarily affect the pleura. PMID- 16716812 TI - Pleural fibrosis. AB - Pleural fibrosis can result from a variety of inflammatory processes. The response of the pleural mesothelial cell to injury and the ability to maintain its integrity are crucial in determining whether normal healing or pleural fibrosis occurs. The pleural mesothelial cell, various cytokines, and disordered fibrin turnover are involved in the pathogenesis of pleural fibrosis. The roles of these mediators in producing pleural fibrosis are examined. This article reviews the most common clinical conditions associated with the development of pleural fibrosis. Fibrothorax and trapped lung are two unique and uncommon consequences of pleural fibrosis. The management of pleural fibrosis, including fibrothorax and trapped lung, is discussed. PMID- 16716813 TI - Imaging of pleural disease. AB - Imaging plays an important role in the diagnosis and subsequent management of patients with pleural disease. The presence of a pleural abnormality is usually suggested following a routine chest x-ray, with a number of imaging modalities available for further characterization. This article describes the radiographic and cross-sectional appearances of pleural diseases, which are commonly encountered in every day practice. The conditions covered include benign and malignant pleural thickening, pleural effusions, empyema and pneumothoraces. The relative merits of CT, MRI and PET in the assessment of these conditions and the role of image-guided intervention are discussed. PMID- 16716815 TI - Pleural manometry. AB - The goals of therapeutic thoracentesis are to remove the maximum amount of pleural fluid to improve dyspnea and to facilitate the diagnostic evaluation of large pleural effusions. Pleural manometry may be useful for immediately detecting an unexpandable lung, which may coexist when any pleural fluid accumulates. Pleural manometry may improve patient safety when removing large amounts of pleural fluid. The basics of pleural space mechanics are discussed as they apply to the normal pleural space and to pleural effusion associated with expandable and unexpandable lung. This article also discusses the instrumentation required to perform bedside manometry, how manometry may decrease the risk of re expansion pulmonary edema when large amounts of fluid are removed, and the diagnostic capabilities of manometry. PMID- 16716814 TI - Pleural ultrasonography. AB - Ultrasonography has achieved acceptance as a routine clinical tool for clinicians managing pleural disease. This article provides an overview of the field of pleural ultrasonography with an emphasis on clinical applicability and procedure guidance. PMID- 16716816 TI - Discriminating between transudates and exudates. AB - The dichotomous classification of pleural fluid as a transudate or an exudate simplifies diagnostic efforts in determining the cause of pleural effusions. Multiple pleural fluid tests are available to discriminate between these two classes of effusions. Tests commonly used in clinical practice depend on the detection in pleural fluid of large-molecular-weight chemicals that enter the pleural space to greater degrees in conditions associated with exudative compared with transudative effusions. Considerable misclassifications can occur with all available testing strategies, so clinicians benefit from adopting a nondichotomous, bayesian approach for interpreting test results. PMID- 16716817 TI - The approach to the patient with a parapneumonic effusion. AB - Parapneumonic effusion is a common clinical problem, and those that go on to develop pleural infection have high morbidity and mortality. The process of pleural infection evolution involves changes in pleural physiology that are increasingly being elucidated and understood. The microbiology of pleural infection has changed over recent years, with clear differences emerging between hospital- and community-acquired infections. Using biochemical surrogates of infection, chest drainage can be undertaken rationally for those who do not respond to antibiotics alone. Recent data suggest that fibrinolytics do not influence outcomes in pleural infection. The optimal type and timing of surgery remain controversial. PMID- 16716818 TI - The spectrum of pleural effusions after coronary artery bypass grafting surgery. AB - Pleural effusions are common after coronary artery bypass grafting (CABG) surgery and can be categorized by time intervals: perioperative (within the first week), early (within 1 month), late (2-12 months), or persistent (after 6 months). The perioperative effusions are usually attributable to diaphragm dysfunction or internal mammary artery harvesting and are typically self-limited. Early effusions are usually attributable to postcardiac injury syndrome and may require corticosteroid treatment. Although late effusions can have multiple causes, persistent effusions are attributable to trapped lung and often require decortication. Diagnostic thoracentesis should be performed for patients with large symptomatic pleural effusions or fever after CABG surgery. The range of management includes observation, therapeutic thoracentesis, corticosteroids, or decortication depending on the cause and course of the effusion. PMID- 16716819 TI - Pleural effusions of extravascular origin. AB - Most pleural effusions are caused by hydrostatic and oncotic pressure imbalance, inflammation or infection, or abnormalities in lymphatic drainage. A select number of effusions are caused by fluid of extravascular origin. Some of these effusions result from complications of treatment, whereas others are a ramification of the underlying disease. The incidence, pathogenesis, clinical presentation, chest radiographic manifestations, pleural fluid analysis, diagnosis, and management are discussed. PMID- 16716820 TI - The undiagnosed pleural effusion. AB - The most common causes for undiagnosed transudative effusions are congestive heart failure and hepatic hydrothorax. Pleural fluid N terminal pro-brain natriuretic peptide levels higher than 1500 pg/mL are virtually diagnostic of congestive heart failure. The most common causes for undiagnosed exudative pleural effusions are malignancy, pulmonary embolism, and tuberculosis. Clinical characteristics of patients with a malignant pleural effusion are symptoms for more than 1 month, absence of fever, blood-tinged pleural fluid, and CT findings suggestive of malignancy. Thoracoscopy is useful to establish the diagnosis of malignancy and tuberculosis. PMID- 16716821 TI - Staphylococcal superantigens of the enterotoxin gene cluster (egc) for treatment of stage IIIb non-small cell lung cancer with pleural effusion. AB - There has been renewed interest in the superantigens as antitumor agents with the discovery of a group of bacterial superantigens known as the enterotoxin gene cluster (egc staphylococcal enterotoxins [SEs]). This article discusses the mechanisms by which egc SEs induce tumor killing and pleurodesis. The application of SE homolog and nucleic acid compositions as vaccines and for treatment of established tumors is reviewed. Finally, the use of native SEs ex vivo intratumorally and intravesicularly administered superantigens against established tumors-is described and the interrelation between superantigen therapy and chemoradiotherapy. PMID- 16716822 TI - Management of malignant pleural mesothelioma. AB - Malignant mesothelioma is increasing in incidence globally and has no known cure. Its unique clinical feature of local infiltration along tissue planes makes it a difficult neoplasm to manage. There have been few randomized controlled trials regarding treatment options, although these have increased in recent years, and results are eagerly awaited. This article summarizes important advances in the management of mesothelioma, especially diagnostic and therapeutic aspects. PMID- 16716823 TI - Pleural disease in lymphangioleiomyomatosis. AB - Pleural disease is a common complication of lymphangioleiomyomatosis (LAM). The incidence and recurrence rates of secondary spontaneous pneumothorax in LAM are the highest among chronic pulmonary disorders. Most patients have at least one pneumothorax before LAM is diagnosed, and pneumothorax is often the sentinel event that leads to the diagnosis. Although early, definitive treatment for recurrent pneumothorax by pleurodesis is recommended, the failure rate for chemical and surgical approaches is high. Chylothorax occurs owing to obstruction of lymphatics by proliferating smooth muscle cells and often requires pleurodesis to control expanding and recurrent effusions. Because of the rarity of the LAM, few data exist on whether the occurrence of pleural complications in LAM accelerates functional decline or portends a worse prognosis. PMID- 16716824 TI - Management of spontaneous pneumothorax. AB - Management of patients with a spontaneous pneumothorax continues to challenge clinicians. Recent guidelines help provide care pathways for these patients and highlight the many areas in need of additional study. Management options for spontaneous pneumothoraces should be selected based primarily upon a patient's clinical status. Observation or pleural air drainage, in selected patients, plays a significant role in patients with primary spontaneous pneumothorax. By contrast, pleural air drainage plays the central role in patients with a secondary spontaneous pneumothorax. Surgically directed recurrence prevention and air leak management are preferred for both primary and secondary spontaneous pneumothorax patients. PMID- 16716825 TI - Adaptive cortical changes and the functional correlates of visuo-motor integration in relapsing-remitting multiple sclerosis. AB - Cortical reorganization has been demonstrated during performance of a motor task in patients with multiple sclerosis. Converging evidence suggests that changes in gray matter volume represent an early hallmark of the disease. We used functional MRI to investigate the role of cortical adaptive mechanisms in maintaining visuo motor function in the face of structural damage. Two cohorts of patients with clinically definite relapsing-remitting multiple sclerosis were compared with healthy controls matched for demographic, motor and cognitive characteristics during the performance of a visuo-motor integration task. Direct comparison between the two groups demonstrated a greater response of the contralateral dorsal premotor cortex and of the ipsilateral superior parietal cortex in relapsing-remitting multiple sclerosis patients. The functional MRI changes in these areas were strongly correlated with decreased gray matter volumes and increased lesion burden, respectively. Our study demonstrated a selective involvement of the parieto-premotor circuitry in a relatively early stage of the disease, which was not influenced by clinical, motor or cognitive variables. Moreover these results confirm the potential for functional recovery and the adaptive role of these areas in the motor reorganization of multiple sclerosis patients. PMID- 16716826 TI - Prolonged intracortical delay of long-latency reflexes: electrophysiological evidence for a cortical dysfunction in multiple sclerosis. AB - Convincing evidence suggests that long-latency reflexes (LLRs) are capable of testing the transcortical sensorimotor reflex arch. By subtracting the sum of the latencies of N20 (afferent branch) and transcranially elicited motor evoked potentials (MEP; efferent branch) from the LLR II latency, the cortical relay time (CRT) can also be obtained, which is alleged to represent the time required for the cortical sensorimotor integration. The aim of the present study was to investigate if a cortical dysfunction occurs in multiple sclerosis (MS). Median nerve somatosensory evoked potentials (SEPs), MEPs and LLRs were recorded from the upper limbs of 23, not severely disabled MS patients in acute phases of the disease. Eighteen age and sex matched healthy volunteers served as controls. N20, MEP, LLR II latencies were measured, and the CRT was calculated for each limb. The statistical comparison between patients and controls was only weakly significant by taking into account conduction times along either the afferent (N20) or the efferent (MEP) pathways. On the contrary, it turned out to be considerably significant if both branches of the transcortical sensorimotor reflex arch, together with the intracortical pathway, were simultaneously tested by means of the LLRs. Moreover, the patients showed a significantly higher CRT compared with that found in the control subjects. These findings are consistent with a prolonged intracortical delay of LLRs in the MS group and suggest the occurrence of conduction velocity slowing and/or synaptic transmission impairment along the sensorimotor intracortical pathway in MS. PMID- 16716828 TI - Release of vesicular Zn2+ in a rat transient middle cerebral artery occlusion model. AB - In the brain, Zn(2+) is stored in synaptic vesicles of a subgroup of glutamatergic nerve terminals. Although it has been reported that this Zn(2+) is released upon the excitation of nerves in vitro, there has been little study of the release of Zn(2+) during ischemia in vivo. Here, using brain microdialysis, the release of vesicular Zn(2+) was investigated in vivo. When the vesicular Zn(2+) was released into the synaptic cleft by a depolarizing stimulation achieved by perfusion with Ringer's solution containing high K(+) (100mM KCl), a significant increase in the extracellular concentration of Zn(2+) could be detected by microdialysis. Then, we investigated the release of vesicular Zn(2+) in a rat transient middle cerebral artery occlusion model using microdialysis. Consequently, the extracellular Zn(2+) level in the cortex increased within 15 min of the start of occlusion and reached a peak at 30 min, which was about twice the basal level. After 30 min, it declined with time returning to the basal level 15 min after reperfusion, which was performed after 60 min of occlusion. The results suggest that vesicular Zn(2+) would be released into the synaptic cleft during brain ischemia in vivo. PMID- 16716827 TI - Regional protein levels of cytosolic phospholipase A2 and cyclooxygenase-2 in Rhesus monkey brain as a function of age. AB - Limited evidence suggests that brain cytosolic phospholipase A(2) (cPLA(2)), which selectively releases arachidonic acid (AA) from membrane phospholipids, and cyclooxygenase-2 (COX-2), the rate-limiting enzyme for AA metabolism to prostanoids, change as a function of normal aging. In this study, we examined the protein levels of cPLA(2) and COX-2 enzymes in hippocampus, frontal pole and cerebellum from young (2-5 years old), middle-aged (8-11 years old) and old (23 years old) male and female Rhesus monkeys. In the cerebellum, cPLA(2) protein level was higher in the young brain as compared to levels seen at both middle aged and old. Similarly, in the frontal pole, the young brain showed a higher level of COX-2 protein as compared to the levels seen at both older ages. For both, once an animal reached 8-11 years of age the levels appeared to remain relatively constant over the next decade. Immunohistochemistry of COX-2 protein within the brain demonstrated no significant change in the localization to neurons within the frontal pole. Qualitatively, a greater number of neurons were positively stained for COX-2 in the young brain than in the aged brain. Based on the previous reports of localization of cPLA(2) and COX-2 at post-synaptic sites in neurons results from the current study suggest that the elevated protein levels of the two enzymes seen in the younger brain is related to the greater potential for synaptic plasticity across multiple neurons as a function of age and that cPLA(2) and COX-2 may be considered as post-synaptic markers. PMID- 16716829 TI - Modulatory effects of d-serine and sarcosine on NMDA receptor-mediated neurotransmission are apparent after stress in the genetically inbred BALB/c mouse strain. AB - Abnormalities of NMDA receptor-mediated neurotransmission are involved in the pathophysiology of schizophrenia, Alzheimer's disease, substance abuse and seizure disorders. The NMDA receptor is implicated in schizophrenia because phencyclidine (PCP), a noncompetitive NMDA receptor antagonist, binds to a hydrophobic domain within the channel, precipitating a schizophreniform psychosis in susceptible persons. Pharmacological, environmental, and genetic variables alter NMDA receptor-mediated neurotransmission. Inbred mouse strains differ in their sensitivity to some of the behavioral effects of MK-801 (dizocilpine), a PCP analogue. The NMDA receptor complex in the BALB/c strain could reflect a unique stoichiometric combination of receptor subunits resulting in a higher proportion of the channels in the open configuration, a higher affinity of MK-801 for its hydrophobic channel domain, and/or a combination of the above. The BALB/c mouse strain, "stressed" mice, and behavioral consequences of MK-801 administration represent models of altered glutamatergic neural transmission. We were interested in examining the effect of stress on the modulatory properties of d-serine and sarcosine. d-Serine is a naturally occurring glycine agonist that modulates the ability of l-glutamate to influence the opening of the NMDA receptor-associated ionophore, and sarcosine is a naturally occurring glycine reuptake inhibitor. The data suggest that 24h after stress, d-serine and sarcosine interact synergistically to reduce MK-801's ability to antagonize electrically precipitated tonic hindlimb extension. Under conditions of stress, modulatory effects of d-serine and sarcosine on the antiseizure effect of MK-801 are observed that are not apparent in the nonstress condition. The results could be relevant to the development of glycinergic interventions for the treatment of neuropsychiatric disorders. PMID- 16716830 TI - The brain 5HTergic response to an acute sound stress in rats with generalized (absence and audiogenic) epilepsy. AB - The brain serotoninergic (5HTergic) system of epileptic subjects can influence their vulnerability to stress. We studied the putative dependency of 5HT neurotransmission parameters on emotional stress, and the presence, types and severity of seizures using rats with genetic generalized (absence and/or audiogenic) epilepsy, of WAG/Rij and Wistar strains. The animals were stressed by exposure to a short aversive noise or left without sound stimulation. Tissue concentrations of 5HT, tryptophan (TRT) and 5-hydroxyindolacetic acid (5HIAA) were assessed by HPLC. The stressor activated the 5HTergic system within thalamus (5HIAA elevated), frontal cortex (5HT, TRT elevated), hypothalamus (increased TRT) in all rats. However, the normal (non-epileptic) rats displayed the highest response in the frontal cortex and the lowest one in the thalamus, as compared to the epileptic rats. Absence-epileptic rats exhibited higher thalamic 5HIAA increase than their controls. Significant correlations existed between propensity of absence epilepsy and 5HTergic parameters measured in the cortex and hypothalamus of absence-epileptic rats. No major difference was found between groups with and without audiogenic epilepsy. The results imply that the stress response depends on the presence of epileptic pathology and the seizure type and severity. The brain 5HT may be involved in the control of the paroxysms and behaviour in absence-epileptic subjects. PMID- 16716831 TI - Magnesium and riboflavin combination therapy following cortical contusion injury in the rat. AB - Previous research has shown that magnesium chloride (MgCl(2)) and riboflavin (B(2)) both significantly improve functional recovery when administered shortly after frontal cortical contusion injury (CCI). The purpose of the present study was to examine the ability of combination treatments of MgCl(2) and B(2) to improve functional outcome following unilateral CCI. One hour post-injury, rats were administered MgCl(2) (1.0 mmol/kg), B(2) (7.5mg/kg), MgCl(2)+B(2) (1 mmol/kg+7.5mg/kg), 1/2 MgCl(2)+1/2 B(2) (0.5 mmol/kg and 3.75 mg/kg), or saline. Two days following CCI rats were tested on a battery of sensorimotor (vibrissae- >forelimb placing and tactile removal test) and motor (staircase test). A regimen of MgCl(2)+B(2) significantly reduced the initial impairment and facilitated the rate of recovery on the tactile removal test and facilitated the rate of recovery on the forelimb placing test. The half-dose combination did not significantly improve functional recovery on the tactile removal test compared to the individual treatments; however, it did improve performance on the forelimb placing test compared to saline treatment. Administration of MgCl(2) improved performance on the placing and tactile removal tests on 2 post-operative days, as did treatment with B(2) on the tactile removal test. The results indicate that the full combination of MgCl(2)+B(2) significantly improved functional recovery to a greater extent than the individual treatments or the low dose combination group on forelimb placing but not on tactile removal. These findings suggest that administration of MgCl(2)+B(2) may provide better therapeutic action than individual treatments. PMID- 16716832 TI - Peripheral and mesencephalic transfer of a synthetic gene for the thymic peptide thymulin. AB - Thymulin is a thymic peptide with antiinflammatory activity in the brain. We constructed a recombinant adenoviral vector, RAd-FTS, expressing a synthetic DNA sequence encoding met-FTS, a biologically active analog of thymulin and used it for peripheral and central gene transfer in rats. Thymulin concentration in serum and brain tissue was determined by bioassay. Reporter gene expression in the substantia nigra (SN) was quantitated by enzymohistochemistry or fluorescence microscopy using an appropriate image analysis software. A single intramuscular injection (10(8) plaque forming units (pfu)/animal) of RAd-FTS in thymectomized rats (nondetectable serum thymulin) induced supraphysiologic serum thymulin levels for at least 110 days (123+/-22 fg/ml versus 598+/-144 fg/ml in intact and vector-injected rats, respectively). Stereotaxic intranigral injection of RAd-FTS induced steady expression levels of met-FTS for at least 90 days, whereas expression of adenovirally transferred reporter genes coding for green fluorescent protein fused to HSV thymidine kinase (GFP-TK)(fus) or E.coli beta galactosidase (beta-gal), declined drastically within a month (% transgene expression in the SN on post-injection day 30 relative to day 2 was: 18, <1 and 125%, for beta-gal, (GFP-TK)(fus) and met-FTS, respectively). We conclude that RAd-FTS constitutes a suitable biotechnological tool for the assessment of peripheral and central thymulin gene therapy in animal models of nigral dopaminergic neurodegeneration induced by pro-inflammatory agents. PMID- 16716833 TI - Evidences for calcium-dependent inactivation of calcium current at the frog motor nerve terminal. AB - Assessment of calcium-dependent inactivation of calcium current in nerve terminals is not feasible due to technical reasons. Perineural measurement of calcium-flow, however, might be utilized as indirect means to evaluate synaptic currents. Using perineural recording from frog neuromuscular junction, supra threshold stimuli applied to motor nerve in paired-pulse manner with varying inter-pulse intervals (5-50 ms) are demonstrated in this study to cause paired pulse depression (PPD) of Ca(2+)-current. PPD of Ca(2+)-flow was reduced at lower extracellular Ca(2+) concentrations, in BAPTA-AM and EGTA-AM treated preparations and after replacing extracellular Ca(2+) with Sr(2+). Using perineural measurement of calcium current as an indirect model to investigate synaptic ionic activity, our findings demonstrate that PPD may be attributed to calcium dependent inactivation of Ca(2+)-current, which may serve as negative feedback in response to massive Ca(2+) entry to motor nerve terminals. A putative sensor of Ca(2+)-current is also proposed in this study. PMID- 16716834 TI - Risperidone attenuates brain damage after focal cerebral ischemia in vivo. AB - Since their introduction, atypical neuroleptic agents have been discovered to have some beneficial effects beyond their effectiveness as neuroleptic drugs. Among these initially unexpected effects are their potential effects as mood stabilizers in bipolar disorder and their efficacy in improving long-term outcome in schizophrenia. These effects recently raised the question whether these drugs may also have some neuroprotective effect in the brain. To examine this matter, in this study we evaluated the neuroprotective effect of risperidone after permanent focal cerebral ischemia. Anaesthetized male C57BL/6j mice were submitted to permanent thread occlusion of the middle cerebral artery (MCA). Risperidone (0.1, 1 or 10 mg/kg) or vehicle was applied intraperitoneally just after permanent ischemia. Twenty-four hours after permanent ischemia, brain injury was evaluated by triphenyltetrazolium chloride staining (TTC). Risperidone (0.1, 1 and 10 mg/kg) showed significant neuroprotection after permanent focal cerebral ischemia. PMID- 16716836 TI - Oxidative and antioxidative capacity in children with cerebral palsy. AB - The superiority of oxidative stress and/or the inadequacy of antioxidant capacity have an important role in disease. Decreased antioxidant availability has been observed in the pathogenesis of many different diseases affecting the brain, such as mitochondrial disorders, cerebral ischaemia and epilepsy. Oxidative and antioxidative status in children with cerebral palsy aged 1-12 years was investigated in this study and compared with healthy controls. Sixty-nine patients with cerebral palsy and 42 controls were enrolled in the study. Lipid peroxidation in the cerebral palsy group was significantly higher than that in the controls (7.54+/-3.64 micromol H(2)O(2)/L and 5.84+/-1.25 micromol H(2)O(2)/L, respectively) (P=0.02). Serum total antioxidant capacity levels were also markedly lower in the CP group than in the control group (1.42+/-0.22 mmol Trolox equiv./L and 1.64+/-0.17 mmol Trolox equiv./L, respectively) (P=0.003). Uric acid and albumin concentrations were lower in the study group than in the control group. Based on these results, we concluded that oxidants were increased and antioxidants were decreased in the cerebral palsy group, and, as a result, the oxidative/antioxidative balance shifted to the oxidative side in children with cerebral palsy. PMID- 16716835 TI - Aversive stimulus attenuates impairment of acquisition in a delayed match to position T-maze task caused by a selective lesion of septo-hippocampal cholinergic projections. AB - Infusion of 192 IgG-saporin (SAP) into the medial septum (MS) of rats selectively destroys cholinergic neurons projecting to the hippocampus and impairs acquisition of a delayed matching to position (DMP) T-maze task. The present study evaluated whether introduction of a mild aversive stimulus 30 min prior to training would attenuate the deficit in DMP acquisition caused by the SAP lesions. Male Sprague-Dawley rats received medial septal infusions of either artificial cerebrospinal fluid or SAP (0.22 microg in 1.0 microl). Fourteen days later, all animals were trained to perform the DMP task. Half of the SAP-treated animals and controls received an intraperitoneal injection of saline each day, 30 min prior to training. Results show that intraperitoneal saline attenuated the impairment in DMP acquisition in SAP lesioned rats. These results suggest that a mild aversive stimulus can attenuate cognitive deficits caused by medial septal cholinergic lesions. PMID- 16716837 TI - Calcineurin inhibitors cause an acceleration of the neurological phenotype in a mouse transgenic for the human Huntington's disease mutation. AB - Calcineurin (CaN) is a Ca(2+)- and calmodulin-dependent protein serine-threonine phosphatase that is thought to play an important role in the neuronal response to changes in the intracellular Ca(2+) concentration. CaN has been implicated in numerous physiological processes including learning and memory. Decreases in CaN expression are thought to be responsible for some of the pathological features seen in brain ischemia, Down's syndrome and Alzheimer's disease. In this study, we examined the possibility of CaN playing a role in the progressive neurological phenotype of the R6/2 mouse of Huntington's disease. We studied the effects of the CaN inhibitors cyclosporin A and FK506 on the progressive neurological phenotype in the R6/2 mouse. We found that an immunosuppressive dose of both drugs dramatically accelerated the main features of the neurological phenotype in R6/2 mice. This was unlikely to be due solely to the immunosuppressive action of these drugs, since treatment with cyclophosphamide, an immunosuppressant drug with a mechanism of action that is not mediated via CaN, did not have deleterious effects on the R6/2 mouse. If anything, cyclophosphamide improved the neurological symptoms in the R6/2 mice. Together, our data suggest a central role for CaN in the deleterious phenotype of the R6/2 mouse. Treatments aimed at preventing the loss of CaN or stimulating its function may be beneficial in the treatment of HD. PMID- 16716838 TI - Inhibitory effect of Cd2+ on glycine-induced chloride current in rat hippocampal neurons. AB - The effects of cadmium (Cd(2+)) on glycine-induced Cl(-) current (I(Gly)) were investigated in acutely dissociated rat hippocampal CA1 neurons using the conventional whole-cell patch-clamp technique in this study. We found that Cd(2+) reversibly and concentration-dependently, reduced the amplitudes of I(Gly), with an IC(50) of 1.27 mM and Hill coefficient of 0.45. The depression of I(Gly) by Cd(2+) was independent of membrane voltage between -60 and +40 mV and did not involve a shift in the reversal potential of the current. A non-competitive inhibition was suggested by a double reciprocal plot of the effects of Cd(2+) on the concentration-response curve of the I(Gly). Since intracellular dialysis with 3mM Cd(2+) failed to modify I(Gly), it was suggested that the site of action of Cd(2+) is extracellular. The suppression of I(Gly) by Zn(2+) was unaffected by 3mM Cd(2+), which indicated that Zn(2+) and Cd(2+) bind to independent sites on glycine receptor. The results show that Cd(2+) decreases I(Gly) in acutely dissociated rat hippocampal neurons and the depression of I(Gly) by Cd(2+) may contribute to worsen the neurotoxicological impairment. PMID- 16716839 TI - Econazole attenuates cytotoxicity of 1-methyl-4-phenylpyridinium by suppressing mitochondrial membrane permeability transition. AB - Defects in mitochondrial function have been shown to participate in the induction of neuronal cell injury. The effect of econazole against the cytotoxicity of 1 methyl-4-phenylpyridinium (MPP(+)) in differentiated PC12 cells was assessed in relation to the mitochondrial membrane permeability changes. Treatment of PC12 cells with MPP(+) resulted in the nuclear damage, decrease in the mitochondrial transmembrane potential, cytosolic accumulation of cytochrome c, activation of caspase-3, increase in the formation of reactive oxygen species (ROS) and depletion of GSH. Econazole (0.25-2.5 microM) inhibited the cytotoxicity of MPP(+) or rotenone. The addition of econazole (0.5 microM) significantly attenuated the MPP(+)-induced mitochondrial damage, elevation of intracellular Ca(2+) level and cell death. However, because of the cytotoxicity, econazole at 5 microM did not attenuate the toxicity of MPP(+). The results show that econazole at the low concentrations may reduce the MPP(+)-induced viability loss in PC12 cells by suppressing the mitochondrial permeability transition, leading to activation of caspase-3 and the elevation of intracellular Ca(2+) levels, which are associated with the increased formation of ROS and depletion of GSH. PMID- 16716841 TI - American Indian adolescents in substance abuse treatment: diagnostic status. AB - The goal of this study was to describe the prevalence of Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) psychiatric disorders among a sample of American Indian (AI) adolescents in residential substance abuse treatment. Data on 89 AI adolescents admitted to a tribally operated residential substance abuse treatment program were collected. Participants reported using a mean of 5.26 substances; 20% percent met DSM-IV criteria for four or more substance use disorders. Marijuana abuse/dependence was the most common substance use disorder (84.3%). Eighty-two percent met criteria for at least one DSM-IV nonsubstance use disorder, the most common of which was conduct disorder (74.2%). These results suggest strong diagnostic parallels between these AI adolescents and their non-AI counterparts who have participated in similar studies, including the considerable diagnostic complexity that was common among the participants in this study. These diagnostic patterns suggest that emerging practices for treating substance-abusing adolescents that have been developed for use with non AI adolescents warrant consideration for use with AI youths. PMID- 16716842 TI - Is conventional wisdom wrong? Coverage for substance abuse treatment under Medicaid managed care. AB - Conventional wisdom suggests that coverage for substance abuse treatment under Medicaid is generally poor, and that access to care may be reduced when control over behavioral health services is given to private health plans, such as those under Medicaid managed care. To examine this premise, this study reports on a cross-sectional comparative survey of state Medicaid managed care programs conducted in the year 2000. Although not all states provided substance abuse benefits under their Medicaid programs, our findings suggest that a majority of states used managed care arrangements to provide substance abuse treatment, with most providing an array of covered services. Most Medicaid behavioral health plans were fully capitated. The number of comprehensive health plans providing substance abuse services was slightly higher than the number of behavioral health carveouts. About half of the waiver programs that covered substance abuse treatment covered methadone maintenance, but waiver programs employing comprehensive health plans were more likely to provide coverage for methadone maintenance. PMID- 16716843 TI - Voucher-based contingent reinforcement of marijuana abstinence among individuals with serious mental illness. AB - Previous studies by our group have used money given contingent on abstinence to reduce drug use by individuals with schizophrenia. In this study, we examined the sensitivity of marijuana use by individuals with serious mental illness to voucher-based contingent reinforcement, which represents the first study to date investigating the efficacy of voucher incentives with this population. This within-subject reversal design consisted of three conditions: 4-week baseline, 12 week voucher intervention, and 4-week baseline. During baseline periods, subjects received 10 US dollars vouchers per urine specimen, independent of urinalysis results. During voucher intervention, only specimens testing negative for marijuana earned vouchers, with total possible earnings of 930 US dollars. Seven adults with schizophrenia or other serious mental illnesses participated in the study. The percentage of marijuana-negative specimens was significantly greater during voucher intervention than during baseline periods. These results provide evidence that marijuana use among individuals with serious mental illness is sensitive to voucher-based incentives and further support the potential feasibility of using voucher-based contingency management to reduce substance abuse in this challenging population. PMID- 16716844 TI - Motivation and readiness for therapeutic community treatment: psychometric evaluation of the Dutch translation of the Circumstances, Motivation, Readiness, and Suitability scales. AB - There is empirical evidence that treatment motivation and readiness are closely linked to retention. Several instruments have been developed to measure these concepts; such instruments can reliably assess the stages of motivation and readiness, and predict treatment retention and outcome. However, limited efforts have been undertaken in Europe to translate those instruments and to determine their reliability and validity. In this study, the psychometric properties of a Dutch translation of the Circumstances, Motivation, Readiness, and Suitability (CMRS) scales by De Leon and Jainchill and the shortened CMR Intake Version are illustrated in a Flemish sample of therapeutic community residents (n = 207). Exploratory factor analyses confirm three CMR subscales, although confirmatory factor analysis reveals only a moderate fit for both the CMRS and the CMR. In contrast, the predictive power of the instrument is strong, specifically for 1 year retention. Significant correlations were found between CMRS measures and age, drug severity, legal referral, and social problems. It is concluded that the Dutch translation of the CMRS and the CMR can be employed as a valid indicator of treatment motivation and readiness, although additional research concerning factorial structures is needed. PMID- 16716845 TI - Acupuncture treatment for opiate addiction: a systematic review. AB - A review of the efficacy of acupuncture as treatment for opiate addiction, covering 33 years of reported literature in western scientific journals, was systematically undertaken. Some abstracts from Chinese language journals were also briefly reviewed. Supportive evidence often came from non-controlled nonblinded methodologies. When well-designed clinical trials (randomized, controlled, single-blind methodologies) were used, there was no significant evidence for acupuncture being a more effective treatment than controls. Some of the current supportive evidence for efficacy came from Chinese journals that have not been translated into English yet. PMID- 16716846 TI - Characteristics of substance abuse treatment programs providing services for HIV/AIDS, hepatitis C virus infection, and sexually transmitted infections: the National Drug Abuse Treatment Clinical Trials Network. AB - Illicit drug users sustain the epidemics of human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS), hepatitis C (HCV), and sexually transmitted infections (STIs). Substance abuse treatment programs present a major intervention point in stemming these epidemics. As a part of the "Infections and Substance Abuse" study, established by the National Drug Abuse Treatment Clinical Trials Network, sponsored by National Institute on Drug Abuse, three surveys were developed; for treatment program administrators, for clinicians, and for state and District of Columbia health and substance abuse department administrators, capturing service availability, government mandates, funding, and other key elements related to the three infection groups. Treatment programs varied in corporate structure, source of revenue, patient census, and medical and non medical staffing; medical services, counseling services, and staff education targeted HIV/AIDS more often than HCV or STIs. The results from this study have the potential to generate hypotheses for further health services research to inform public policy. PMID- 16716847 TI - Is alcoholism typology a predictor of both Alcoholics Anonymous affiliation and disaffiliation after treatment? AB - Twelve Steps (TS) has demonstrated effectiveness; induction into Alcoholics Anonymous (AA) is a primary objective of TS and is a pivotal mechanism explaining its effectiveness. However, evidence suggests that, after treatment, dropout from AA is high. This study investigated whether alcohol problem severity predicted both AA affiliation and disaffiliation among clients receiving TS. This study of a Project MATCH sample included 453 alcohol-dependent clients randomly assigned to TS who reported AA attendance during treatment. Greater alcohol problem severity predicted AA attendance; opposite to prediction, less alcohol-impaired clients were more than twice as likely to discontinue AA attendance after treatment. When sustained AA attendance is desired, the evaluation of client pretreatment alcohol involvement may be useful for identifying potential AA dropout after TS treatment. Findings also indicate that, among treatment-seeking problem drinkers, AA dropout and disaffiliation are distinct, albeit correlated, constructs that require future investigation. PMID- 16716848 TI - Safety, efficacy, and tolerability of nelfinavir-containing antiretroviral therapy for patients coinfected with HIV and hepatitis C undergoing methadone maintenance. AB - The safety, efficacy, and tolerability of nelfinavir (NFV)-containing antiretroviral therapy were evaluated in patients coinfected with HIV and hepatitis C undergoing methadone maintenance at an urban outpatient opioid treatment program serving a minority adult population. Eligibility covered methadone-maintained patients coinfected with HIV and hepatitis C who had received or were currently receiving NFV. The yield was 51 case patients. Parameters examined looked into safety, efficacy, and tolerability. Nelfinavir was discontinued in 2 patients for liver function abnormalities but resumed in 1 patient. One patient developed laboratory abnormalities during NFV therapy that were not present before NFV therapy; in 12 case patients, pre-NFV therapy liver function abnormalities resolved completely during NFV therapy. There was a statistically significant increase in CD4 count during NFV therapy. Viral load decreased or was unchanged in 10 case patients and increased in 8, of whom 5 had a CD4 count increase during NFV therapy. Three patients had diarrhea and 4 patients had constipation. Nelfinavir was not discontinued -- neither was dose adjusted -- in any of these patients. Patients who had received NFV > or =36 months had a smaller increase in mean methadone dose as compared with patients who had received NFV <36 months. The results show that NFV is safe, efficacious, and well tolerated. PMID- 16716849 TI - A practical system for monitoring the outcomes of substance use disorder patients. AB - Outcomes monitoring and management are receiving mounting attention because of increased emphasis on health care accountability and cost containment. Efficient, practical outcomes monitoring systems (OMSs) are crucial if health care system performance is to be determined and effective/cost-effective treatments are to be identified, but such practical monitoring systems generally are lacking. This article describes the features of such a system for monitoring the care received by, and the substance use and psychosocial outcomes of, patients treated for substance use disorders (SUDs) in the Department of Veterans Affairs (VA). In contrast to a 15-21% follow-up rate achieved by VA SUD program staff under a previously mandated system-wide monitoring system, the monitoring system used in this project achieved a 67% follow-up rate without paying patients for their participation. We provide data on patient characteristics and treatment outcomes, estimate the cost of implementing this type of monitoring system on a broad scale, and provide recommendations for OMSs in other large health care organizations. PMID- 16716850 TI - Alcohol relapse repetition, gender, and predictive validity. AB - Earlier exploratory work on a scoring algorithm for the Reasons for Drinking Questionnaire. Exploratory findings from the Reasons for Drinking Questionnaire presented a number of interesting findings, but were limited by a fairly low consistency in type of relapse between the first relapse and the second relapse (63%). This scoring algorithm objectively classifies alcohol relapses into one of three types (negative affect, social pressure, or craving/cued). While examining gender differences in the type of first relapse, evidence indicating that relapses were more consistent for men (81%) than for women (44%) was uncovered. For initial posttreatment relapses, women were more likely to have negative affect relapses, and men were more likely to have social pressure relapses. For men, negative affect relapses were predicted by the Beck Depression Inventory score. For women, negative affect relapses were predicted by the Alcohol Dependence Scale score, and craving/cued relapses were predicted by situational craving. PMID- 16716851 TI - Depression among heroin users: 12-Month outcomes from the Australian Treatment Outcome Study (ATOS). AB - A cohort comprising 495 heroin users were interviewed for the Australian Treatment Outcome Study and were re-interviewed at 12-month follow-up. The rate of current major depression declined significantly from 26% to 11% for the follow up period. Those with current major depression on follow-up experienced fewer total days in treatment, but engaged in more treatment episodes. In comparison to those without depression, depressed individuals had less exposure to methadone/buprenorphine maintenance and residential rehabilitation for the follow up period, but spent more time in detoxification. Those with current major depression on follow-up also reported heavier heroin and other drug use, more risk-taking behaviors, poorer physical health, and greater psychopathology than those without a diagnosis of current major depression. Although caution in interpreting these relationships is advised given the potential for confounding by client characteristics, the findings of this study illustrate the need to consider depression in the treatment of heroin dependence. PMID- 16716852 TI - Early adoption of buprenorphine in substance abuse treatment centers: data from the private and public sectors. AB - The recent approval of buprenorphine for the treatment of opiate dependence offers an opportunity to analyze innovation adoption in community-based treatment. Using data collected from national samples of 299 privately funded and 277 publicly funded treatment centers, this research examines buprenorphine adoption using baseline data collected between 2002 and 2004 as well as follow-up data collected 12 months later. Private centers were significantly more likely than public centers to report current use of buprenorphine. The baseline data indicated that early adoption was positively associated with center accreditation, physician services, availability of detoxification services, current use of naltrexone, and the percentage of opiate-dependent clients. Multivariate analyses of follow-up data suggest that adoption was greater in accredited centers, for-profit facilities, organizations offering detoxification services, and naltrexone-using centers. Future research should continue to monitor the extent to which buprenorphine is adopted in these settings. PMID- 16716853 TI - Evidence-based nursing and mental health concerns: a cross-national discussion. PMID- 16716854 TI - College women's experience of stalking: mental health symptoms and changes in routines. AB - BACKGROUND: Stalking is a serious public health and societal concern affecting many college women. PURPOSE: The purpose of this study was to explore college women's experiences of stalking. The specific aims were to compare victims and nonvictims on physical and mental health indicators and to identify lifestyle changes made in response to being stalked. METHODS: In this cross-sectional design, 601 women from two universities completed a stalking questionnaire, a mental health screening tool, and an injury checklist. Data analysis included frequencies, multivariate analysis of variance, analysis of variance, and chi(2) analysis. RESULTS: A quarter of the sample reported experiencing stalking, most often by an intimate or dating partner. Individuals who reported experiencing stalking reported significantly more mental health symptoms and lower perceived physical health status than individuals who did not. Victims reported changing routines, behaviors, and activities. CONCLUSIONS: Psychiatric nurses must be knowledgeable about stalking and its impact on health. Nurses can provide support, services, and community referrals. PMID- 16716855 TI - Emergency department visits for suicidality in three hospitals. AB - The purposes of this study were to describe suicidal persons who come to the emergency department (ED) and to examine the relationship between clinical and health service characteristics and decisions regarding post-ED care. Data were collected from three hospital EDs by means of a retrospective review of records. During the 1-month study period, 163 ED visits were for suicidal ideation (f = 110) or behavior (f = 53). The mean age of the patients was 36.5 years (range = 5 87 years); 51% were female patients. Clinical decisions about post-ED care tended to be cautious, regardless of a patient's level of suicidality, with 71% of patients either transferred for psychiatric evaluation or admitted to the psychiatric unit. Emergency department staff tended to be slightly more conservative than mental health professionals, but the difference in their decisions about disposition was not significant. PMID- 16716857 TI - Hopelessness and suicide risk emerge in psychiatric nurses suffering from burnout and using specific defense mechanisms. AB - Burnout in psychiatric nurses is a phenomenon of great concern. We conducted an investigation of 120 nurses working in the psychiatric, general medicine/rehabilitation, and critical care/surgery wards to explore the correlations of level of burnout and defense mechanisms with hopelessness. The nurses were administered the Maslach Burnout Inventory, Gleser and Ihilevich's Defense Mechanisms Inventory, and Beck's Hopelessness Scale. The results indicate that burnout and some of the defense mechanisms predicted the level of hopelessness -- a predictor of suicide risk. PMID- 16716856 TI - Older adults in the SeniorWISE study at risk for mild cognitive impairment. AB - As part of a larger clinical trial, SeniorWISE (Wisdom Is Simply Exploration), this study provides baseline affective, cognitive, and functional ability data and reports on the likely incidence of mild cognitive impairment (MCI) in a tri ethnic community sample of older adults (N = 265). Seventy-eight individuals had memory complaints, whereas 105 had none. Of the complainants, 32 had normal memory function and 46 had poor memory performance. Among those without memory complaints, 42 had no memory impairment and 63 had poor memory performance. Forty six individuals (17%) met the criteria of poor everyday memory functioning and memory complaints whereas 81 (31%) would be considered to be at risk based on other MCI criteria. This pattern of results suggests that those with declining memory are less aware of their deficits in activities of daily living that their actual performance would suggest are occurring. PMID- 16716858 TI - Relationships among trait, situational, and comparative optimism: clarifying concepts for a theoretically consistent and evidence-based intervention to maximize resilience. AB - Research evidence identifies trait optimism as the strongest individual predictor of adolescent resilience when framed within Lazarus's theory of stress and coping. As a precursor to the development of an intervention to maximize resilience, this study clarifies the concept of optimism. Relationships among trait, situational, and comparative optimism were explored. The sample (N = 132) consisted of rural adolescents between 14 and 18 years old who were attending public school. The results demonstrate weak correlations, suggesting that adolescents' perceptions of situations may be amenable to an intervention to maximize resilience but that adolescents also do not expect to experience health or environmental problems. Implications for developing an intervention are discussed. PMID- 16716860 TI - Creating an organizational diversity vision: goals, outcomes, and future directions of the International Society of Psychiatric Nurses. AB - The aim of this article was to chronicle the development and status of a diversity vision within a psychiatric nursing organization. Organizational activities to support diversity and cultural competence, along with challenges to a full integration of these values, are presented. Psychiatric nurses need to be engaged in the process of developing personal and organizational cultural competence to eliminate barriers that contribute to disparities in effective mental health care. PMID- 16716859 TI - Re: Group therapy for survivors of childhood sexual abuse. PMID- 16716861 TI - Down the drain: the cost of medications wasted in hospice. PMID- 16716862 TI - What is palliative care performed in certified palliative care units in Japan? PMID- 16716863 TI - Percutaneous vertebroplasty as a palliative measure in the setting of chronic infection. PMID- 16716864 TI - Metastatic breast cancer causing gas gangrene. PMID- 16716865 TI - Subanesthetic ketamine for cancer pain and scientific rigor in cancer pain trials. A reply to Jackson et al. PMID- 16716866 TI - Ketamine and HTLV-1 myelopathy: NMDA blockade and immunomodulation? PMID- 16716868 TI - Fentanyl patch sufficient analgesia for only one day. PMID- 16716869 TI - Reversal of morphine with naloxone precipitates haloperidol-induced extrapyramidal side effects. PMID- 16716870 TI - A randomized, double-blind, placebo-controlled study of intrathecal ziconotide in adults with severe chronic pain. AB - Safety and efficacy data from a study of slow intrathecal (IT) ziconotide titration for the management of severe chronic pain are presented. Patients randomized to ziconotide (n = 112) or placebo (n = 108) started IT infusion at 0.1 microg/hour (2.4 microg/day), increasing gradually (0.05-0.1 microg/hour increments) over 3 weeks. The ziconotide mean dose at termination was 0.29 microg/hour (6.96 microg/day). Patients' baseline Visual Analogue Scale of Pain Intensity (VASPI) score was 80.7 (SD 15). Statistical significance was noted for VASPI mean percentage improvement, baseline to Week 3 (ziconotide [14.7%] vs. placebo [7.2%; P = 0.036]) and many of the secondary efficacy outcomes measures. Significant adverse events (AEs) reported in the ziconotide group were dizziness, confusion, ataxia, abnormal gait, and memory impairment. Discontinuation rates for AEs and serious AEs were comparable for both groups. Slow titration of ziconotide, a nonopioid analgesic, to a low maximum dose resulted in significant improvement in pain and was better tolerated than in two previous controlled trials that used a faster titration to a higher mean dose. PMID- 16716872 TI - The effect of a physical exercise program in palliative care: A phase II study. AB - The purpose of this pilot study was to assess the effects of a physical exercise program on physical performance and quality of life (QOL) in a population with incurable cancer and a short life expectancy. Thirty-four patients participated in a 50-minute group exercise program twice a week for 6 weeks. Physical performance was measured by three tests: "6-minute walk test," "timed repeated sit to stand," and "functional reach." Fatigue was measured by the Fatigue Questionnaire. QOL was assessed by the European Organization for Research and Treatment of Cancer Core Quality of Life Questionnaire. The outcome variables were assessed before and after the intervention. The walk length increased and the "timed repeated sit to stand" was reduced (P < 0.05). Emotional functioning improved and physical fatigue was reduced (P < 0.05). Physical exercise seems to be a feasible way to improve well-being among patients with incurable cancer. Future randomized trials are needed to confirm the results. PMID- 16716871 TI - Caregiving at the end of life: Perceptions of health care quality and quality of life among patients and caregivers. AB - This study explored the association between perceptions of health care quality and quality of life in patients with advanced metastatic cancer and their informal caregivers (n=39). Patients' and caregivers' perceptions of health care quality, mental health, health-related quality of life, symptoms, and burden were measured. The key findings included the following: 1) patients' mental health and depression scores correlated with those of caregivers, suggesting that the mental health of patients and their caregivers are associated; 2) patients and caregivers shared similar perceptions regarding health care quality; 3) the presence of depression in caregivers correlated with caregivers being less satisfied with the health care being given to their patients (this correlation did not exist for patients, a finding that may be due in part to the protective buffering effect that caregivers provide their patients as illness progresses); and 4) a modified Primary Care Assessment Survey, originally designed for primary care patients, was a useful measure of health care assessment for both patients and caregivers. These data suggest that patients with advanced disease and their caregivers share similar perceptions and evolve as a "unit of care," and caregivers, as unique and important members of the patient's health care team, are also in need of care. When depressed, caregivers may unilaterally lose trust by becoming less satisfied with the quality of health care being provided to their patients. PMID- 16716873 TI - Status of cancer pain in Hanoi, Vietnam: A hospital-wide survey in a tertiary cancer treatment center. AB - Little is known about the prevalence of cancer pain in many developing countries. We report a hospital-wide survey of pain in a tertiary cancer center in Hanoi, Vietnam. All inpatients and outpatients age 18 years or older were approached for participation in the study. Data were collected using the Brief Pain Inventory. Results showed a 70% response rate. Prevalence of moderate to severe pain was 50% (89/178), with 23% reporting severe ratings of pain at its "worst" and 33% reporting severe impairment in their ability to work due to pain. Only 1% and 40% reported total and partial pain relief from medications, respectively. This study is among the first to provide a representative view of pain in a tertiary cancer treatment center in Hanoi, Vietnam. The findings provide empirical support for the need for better programmatic efforts to improve relief of cancer pain in developing countries, including Vietnam. PMID- 16716875 TI - The influence of awareness of terminal condition on spiritual well-being in terminal cancer patients. AB - We developed a Spirituality Transcendence Measure (STM) and studied whether awareness of terminal illness affects spiritual well-being in terminal cancer patients. Three sources of spiritual transcendence--the situational, the moral and biographical, and the religious aspect--were assessed in the STM. Cronbach's alpha of the STM was 0.95, and the principle axis factor analysis extracted only one factor. Thirty-seven terminal cancer patients with male predominance (59.5%) were studied. Awareness of terminal illness was associated with a higher total STM score (Z = -2.21, P = 0.027), along with the individual scores for each of the three transcendences (Z = -2.39, P = 0.017; Z = -2.71, P = 0.007; and Z = 1.96, P = 0.050). Acceptance of death was associated with a higher situational score (Z = 2.01, P = 0.046) and a higher religious score (Z = -2.27, P = 0.023). Announcement of testament was associated with a higher situational score (Z = 2.30, P = 0.021). We conclude that awareness of terminal illness is associated with spiritual well-being. Telling the complete truth is necessary even when dealing with terminal conditions. PMID- 16716874 TI - Pediatric pain prevalence and parents' attitudes at a cancer hospital in Jordan. AB - There has been little research on implementation of pediatric pain programs. These studies are part of a project to develop such a program for the King Hussein Cancer Centre in Jordan. Study 1 captured information on pain prevalence in 35 children using chart reviews and parent/child interviews to establish baseline pain burden. Forty-seven percent of children had pain at the time of interview; 11% had "a lot" of pain and only 22% received analgesics. Twenty-two parents were interviewed in Study 2 to identify attitudes toward pain management. Thematic analysis revealed six themes: 1) pain can and should be managed; 2) God's will; 3) parent's worst pain was emotional pain due to child's diagnosis; 4) belief that their presence could ameliorate their child's pain; 5) desire for shared decision making; and 6) the child's responsibility to express pain. These study results were used to inform the action research approach in the overall project. PMID- 16716876 TI - Drug utilization review on a tertiary palliative care unit. AB - Drugs are indispensable for the management of symptoms in palliative care patients, and account for a significant proportion of expenditures on a Tertiary Palliative Care Unit (TPCU). Drug expenditures for Edmonton's TPCU increased by 40% in 2002 compared to 2001. Fifty-five percent of the increase was attributable to injectable fentanyl, oral and injectable ondansetron, and total parenteral nutrition (TPN). As there was no increase in the unit cost of these drugs between 2001 and 2002, the increased expenditures reflected increased utilization. The hypothesis of this study was that the increased utilization of these drugs reflected appropriate prescribing. The objective was to compare the indications for prescribing these drugs in 2002 against evidence- and consensus-based criteria. Patients who received these drugs while admitted to the TPCU from January 1 to December 31, 2002 were identified through the pharmacy database. Evidence- and consensus-based criteria for drug utilization were developed. Prescribing indications were retrospectively compared against the criteria. Drug prescriptions were categorized as follows: (1) meeting criteria, (2) not meeting criteria, or (3) uncertain. The drugs under study were prescribed during 48 out of 234 admissions to the TPCU in 2002. Prescriptions for fentanyl met criteria in 26 of 29 cases. Indications were unsuccessful therapy with morphine, hydromorphone, and oxycodone (20), requirement for rapid titration from fentanyl patch (5), renal failure (2), and sublingual administration for breakthrough pain (1). Prescriptions for ondansetron met criteria in 19 of 21 cases. Indications were nausea refractory to metoclopramide and dexamethasone (13), and nausea related to radiotherapy or chemotherapy (6). Prescriptions for TPN met criteria for initiation in only one of five cases. However, in all cases, TPN had been started prior to admission. In cases where death was considered imminent, TPN was continued pending consultation with the patient and family regarding discontinuation. These data indicate that the increased prescribing of fentanyl and ondansetron on the TPCU satisfied evidence- and consensus-based criteria in most cases, apparently justifying the associated increase in drug expenditures. This type of analysis may be useful whenever increased drug utilization requires review. A cost effectiveness analysis would be the next step in evaluating the costs vs. the benefits. The issue of discontinuing TPN in palliative care patients requires further investigation. PMID- 16716877 TI - A comparison of the abuse liability of tramadol, NSAIDs, and hydrocodone in patients with chronic pain. AB - Concern about abuse/dependence in chronic pain patients taking opioid analgesics may lead to undertreatment of pain, yet little is known about the prevalence of abuse/dependence in these patients and how it differs among analgesic agents. The objective of this study was to assess the prevalence of tramadol abuse compared to nonsteroidal anti-inflammatory drugs (NSAIDs) and hydrocodone-containing analgesics in patients with chronic noncancer pain (CNP). The study had three arms. The first arm consisted of subjects prescribed tramadol alone; the second of subjects randomized to either NSAIDs or tramadol; and the third of subjects randomized to hydrocodone or tramadol. Each investigator received two boxes of prescriptions randomized so that one in every four prescriptions was for tramadol. Upon deciding on the therapeutically appropriate arm, the physician selected the appropriate box, opened the next envelope and completed the enclosed prescription. After the initial randomization, physicians could prescribe whatever medication was therapeutically appropriate. A total of 11,352 subjects were enrolled. Up to nine interviews using a structured questionnaire were conducted over a 12-month period. An algorithm called the "Abuse Index" was developed to identify subjects who were abusing the drug. The primary components of the index were increasing dose without physician approval, use for purposes other than intended, inability to stop its use, and withdrawal. The percent of subjects who scored positive for abuse at least once during the 12-month follow up were 2.5% for NSAIDs, 2.7% for tramadol, and 4.9% for hydrocodone. When more than one hit on the algorithm was used as a measure of persistence, abuse rates were 0.5% for NSAIDs, 0.7% for tramadol, and 1.2% for hydrocodone. Thus, the results of this study suggest that the prevalence of abuse/dependence over a 12 month period in a CNP population that was primarily female was equivalent for tramadol and NSAIDs, with both significantly less than the rate for hydrocodone. PMID- 16716879 TI - Epidemiology of crystal arthropathy. AB - Gout is an inflammatory arthritis mediated by the crystallization of uric acid within the joints and often is associated with hyperuricemia. Data suggest that the overall disease burden of gout remains substantial and may be increasing. Identifying and characterizing modifiable risk factors for gout is a major step in preventing and managing this painful condition. As more scientific data on the risk factors and comorbidities of gout become available, their integration into gout prevention and care strategies may become essential. This article reviews the relevant epidemiologic data, with a focus on recent progress and data on other crystal arthropathies. PMID- 16716880 TI - Hyperuricemia and associated diseases. AB - After introduction of urate-lowering therapy, asympotomatic hyperuricemia was treated with allopurinol or uricosuric agents in the belief that hyperuricemia and/or gout caused chronic kidney disease. Epidemiologic studies in the 1970s, however, failed to confirm the view that hyperuricemia and gout were independent risk factors for chronic kidney disease. As a result, urate-lowering pharmacotherapy is generally not recommended at the present time in the management of asymptomatic hyperuricemia even though recent epidemiological, experimental, and clinical studies have prompted reexamination of a causal role for hyperuricemia (with or without gout) in chronic kidney disease as well as other important disorders including cardiovascular disease, hypertension, and metabolic syndrome. The issue of such a role remains unresolved and this article reviews the current status of the relationship between hyperuricemia and associated disorders. PMID- 16716881 TI - Gout: update on some pathogenic and clinical aspects. AB - Pathogenesis of gout inflammation remains unknown, but recent advances have been made with respect to initiation, amplification, and self-limitation. Direct crystal-cell membrane contact leads to cell activation, involving membrane associated molecules. Resolution of acute gout inflammation is a mechanism that is controlled by monocyte-macrophage switch, which results in the loss of cytokine production capability and, conversely, the ability to produce anti inflammatory molecules. MRI and ultrasound findings provide preliminary data that are not yet used in clinical trials. Diagnosis and management recommendations are missing, but this gap will be filled soon with the upcoming European League Against Rheumatism Task Force's recommendations on gout. PMID- 16716882 TI - Renal urate transport. AB - Serum uric acid is determined by a balance between production and renal excretion. Luminal reabsorption of urate by the proximal tubule from the glomerular ultrafiltrate involves coupling between sodium-anion cotransport and urate-anion exchange. Apical sodium-coupled cotransport of lactate, ketoacids, nicotinate, and pyrazinoate increases intracellular levels of these anions in proximal tubular cells, stimulating the apical absorption of luminal urate via anion exchange. Hyperuricemia occurs when plasma levels of these anions increase; for example, hyperuricemia is a well-recognized concomitant of lactic acidosis and ketoacidosis. Relevant developments in the molecular and renal physiology of urate homeostasis are reviewed. PMID- 16716883 TI - Treatment of crystal arthropathy--history and advances. AB - The history of gout and the many distinguished historical figures who have suffered the agonies of this crystal deposition disorder have claimed the attention of medical historians like no other disease. Its treatment with uric acid lowering drugs became a twentieth century paradigm for the successful management and prevention of a chronic rheumatic disease, but the colorful history of the treatment of gout and crystal deposition disorders stretches back over 4,000 years. PMID- 16716884 TI - Clinical trials in crystal arthropathy. AB - A few clinical trials have evaluated therapeutic agents for crystal-associated arthropathy. Most of the studies are uncontrolled and observational. Management of patients who have acute crystal arthropathies usually is symptomatic with long term management depending on crystal composition. In trials of gout, studies focus on acute symptomatic treatment, foregoing chronic management, which is aimed at reducing the concentration of serum urate. In those who have calcium crystals, however, there is no definitive or effective long-term treatment in chronic gout. The xanthine oxidase inhibitor and uricosurics are the agents used most commonly. Newer compounds in clinical trials show promise as effective and safe therapeutic options. PMID- 16716885 TI - Calcium crystal deposition diseases: update on pathogenesis and manifestations. AB - Basic calcium phosphate (BCP) and calcium pyrophosphate dihydrate crystals are the most common types of pathologic calcium-containing crystals. Although these crystals long have been associated with a variety of rheumatic syndromes, recent evidence implicates BCP crystals in the pathogenesis of breast cancer and atherosclerosis. Although understanding of molecular mechanisms involved in generating these pathologic effects has been advanced significantly in recent years, they still are understood incompletely. Such advances are essential to the ongoing search for effective therapies for crystal-associated diseases. PMID- 16716887 TI - Calcium deposition and associated chronic diseases (atherosclerosis, diffuse idiopathic skeletal hyperostosis, and others). AB - Extracellular matrix mineralization or calcification occurs in many pathologic conditions, including atherosclerosis, medial wall calcification, diffuse idiopathic skeletal hyperostosis, and chondrocalcinosis. Vascular wall calcification is the most common and involves two mechanisms: passive calcification resulting from breakdown of the protection system and active calcification resulting from transdifferentiation of mesenchymal cells in the vascular wall to bone. Although reports are conflicting, several matrix proteins are identified as protective factors against dystrophic calcification in nonosseous tissues. Serum matrix Gla protein may be a marker of osteometabolic syndromes that cause hyperostosis and plays a role in Milwaukee shoulder syndrome. PMID- 16716886 TI - Calcium crystal deposition and osteoarthritis. AB - Calcium crystals are common and under-recognized participants in osteoarthritis. Excellent evidence supports two hypotheses explaining the relationship between calcium crystal deposition and osteoarthritis. There is ample support for the theory that calcium crystals cause or worsen osteoarthritis and equally compelling evidence to support the theory that osteoarthritis causes or worsens calcium crystal formation. Further research in this area will improve understanding of the pathogenesis of these conditions and should lead to the development of effective therapy for all types of degenerative arthritis. PMID- 16716888 TI - Imaging features of crystal-induced arthropathy. AB - Crystal-induced arthropathies constitute a spectrum of inflammatory arthritides that is induced by cellular reaction to crystal deposition in and around joints. A variety of microcrystals may be deposited and can induce an inflammatory response. The three most common types of crystal-induced arthropathy are gout, calcium pyrophosphate dihydrate deposition disease, and calcium hydroxyapatite deposition disease. Each has a characteristic clinical presentation, crystal type that may be aspirated from affected tissues, and radiographic appearance. Each of these entities may occur as a primary abnormality or secondary to an underlying disorder. Sometimes these diseases may coexist in the same joint or individual. Imaging frequently plays a crucial role in the diagnosis of crystal-induced arthropathies and may help to monitor disease progression and treatment response. PMID- 16716889 TI - Sphingolipid signaling and redox regulation. AB - Sphingolipids including ceramide and its derivatives such as ceramide-1 phosphate, glycosyl-ceramide, and sphinogosine (-1-phosphate) are now recognized as novel intracellular signal mediators for regulation of inflammation, apoptosis, proliferation, and differentiation. One of the important and regulated steps in these events is the generation of these sphingolipids via hydrolysis of sphingomyelin through the action of sphingomyelinases (SMase). Several lines of evidence suggest that reactive oxygen species (ROS; O2-, H2O2, and OH-,) and reactive nitrogen species (RNS; NO, and ONOO-) and cellular redox potential, which is mainly regulated by cellular glutathione (GSH), are tightly linked to the regulation of SMase activation. On the other hand, sphingolipids are also known to play an important role in maintaining cellular redox homeostasis through regulation of NADPH oxidase, mitochondrial integrity, and antioxidant enzymes. Therefore, this paper reviews the relationship between cellular redox and sphingolipid metabolism and its biological significance. PMID- 16716890 TI - Protein thiol oxidation in health and disease: techniques for measuring disulfides and related modifications in complex protein mixtures. AB - Oxidant species are known to contribute to disease and dysfunction in biological systems. However, evidence has been progressively accumulating that demonstrates a more fundamental role for many oxidant species in the regulation of everyday function of healthy cells. Redox dependent signaling events involving the post translational, oxidative modification of proteins has now been accepted as an important regulatory process, although the full extent of such mechanisms is yet to be determined. Some protein cysteinyl thiols are known to be susceptible to a number of redox-dependent modifications, including an interchange between the reduced thiol and several different oxidized disulfide states. Here, the role of oxidants as regulatory entities is reviewed, as are the many different ways protein disulfide formation can be analysed in complex protein mixtures. This includes an overview of many of the Proteomic strategies that can be used to identify proteins that form disulfides when pro-oxidizing conditions arise in cells, as well as related methods for studying intermediates that may precede disulfide formation. PMID- 16716891 TI - Lack of CuZnSOD activity: a pointer to the mechanisms underlying age-related loss of muscle function, a commentary on "absence of CuZn superoxide dismutase leads to elevated oxidative stress and acceleration of age-dependent skeletal muscle atrophy". PMID- 16716892 TI - Subcellular localization of tyrosine-nitrated proteins is dictated by reactive oxygen species generating enzymes and by proximity to nitric oxide synthase. AB - Using high-resolution immuno-electron microscopy the steady-state subcellular distribution of tyrosine-nitrated proteins in different cells and tissues was evaluated. In quiescent eosinophils and neutrophils in the bone marrow intracellular nitrated proteins were mainly restricted to the peroxidase containing secretory granules. The inducible nitric oxide synthase (iNOS) was expressed in the same granules. Proteins nitrated on tyrosine residues were also abundant in the cytosol of circulating erythrocytes. In the vasculature, nitrated proteins were mainly located in mitochondria and endoplasmic reticulum of the endothelial cells, fibroblasts, and smooth muscle cells. Endogenous nitrated proteins were also found in chondrocytes in cartilage, where it was typically associated with the cytoplasmic interface of the endoplasmic reticulum membrane. Nitrated proteins were also prominent in the peroxisomes of liver hepatocytes and of secretory cells in the lacrimal gland. Challenge of mouse dendritic cells with lipopolysaccharide induced iNOS protein expression in cytosol and peroxisomes and was associated with an increased 3-nitrotyrosine formation in cytosol, mitochondria, and peroxisomes. These data indicate that nitric oxide-dependent protein tyrosine nitration is a physiologically relevant process localized within specific subcellular compartments in close proximity to iNOS and to enzymes capable of peroxidative chemistry and reactive oxygen species production. PMID- 16716893 TI - Ozone induces oxidative stress in rat alveolar type II and type I-like cells. AB - Ozone is a highly reactive gas present in urban air, which penetrates deep into the lung and causes lung injury. The alveolar epithelial cells are among the first cell barriers encountered by ozone. To define the molecular basis of the cellular response to ozone, primary cultures of rat alveolar type II and type I like cells were exposed to 100 ppb ozone or air for 1 h. The mRNA from both phenotypes was collected at 4 and 24 h after exposure for gene expression profiling. Ozone produced extensive alterations in gene expression involved in stress and inflammatory responses, transcription factors, antioxidant defenses, extracellular matrix, fluid transport, and enzymes of lipid metabolism and cell differentiation. Real-time reverse transcription-polymerase chain reaction and Western blot analysis verified changes in mRNA and protein levels of selected genes. Besides the increased stress response, ozone exposure downregulated genes of cellular differentiation. The changes were more prominent at 4 h in the type I like phenotype and at 24 h in the type II phenotype. The type I-like cells were more sensitive to ozone than type II cells. The genome-wide changes observed provide insight into signal pathways activated by ozone and how cellular protection mechanisms are initiated. PMID- 16716894 TI - Arachidonic acid suppresses growth of human lung tumor A549 cells through down regulation of ALDH3A1 expression. AB - Expression of aldehyde dehydrogenase 3A1 (ALDH3A1) in certain normal and tumor cells is associated with protection against the growth inhibitory effect of reactive aldehydes generated during membrane lipid peroxidation. We found that human lung tumor (A549) cells, which express high levels of ALDH3A1 protein, were significantly less susceptible to the antiproliferative effects of 4 hydroxynonenal compared to human hepatoma HepG2 or SK-HEP-1 cells that lack ALDH3A1 expression. However, A549 cells became susceptible to lipid peroxidation products when they were treated with arachidonic acid. The growth suppression of A549 cells induced by arachidonic acid was associated with increased levels of lipid peroxidation and with reduced ALDH3A1 enzymatic activity, protein, and mRNA levels. Furthermore, arachidonic acid treatment of the A549 cells resulted in an increased expression of peroxisome proliferator-activated receptor gamma (PPARgamma), whereas NF-kappaB binding activity was inhibited. Blocking PPARgamma using a selective antagonist, GW9662, prevented the arachidonic acid-mediated reduction of ALDH3A1 expression as well as the growth inhibition of A549 cells, suggesting the central role of PPARgamma in these phenomena. The increase in PPARgamma and the reduction in ALDH3A1 were also prevented by exposing cells to vitamin E concomitant with arachidonic acid treatment. In conclusion, our data show that the arachidonic acid-induced suppression of A549 cell growth is associated with increased lipid peroxidation and decreased ALDH3A1 expression, which may be due to activation of PPARgamma. PMID- 16716895 TI - Potential artifacts in the measurement of DNA deamination. AB - Attack on DNA by some reactive nitrogen species results in deamination of adenine and guanine, leading to the formation of hypoxanthine and xanthine, respectively. Published levels of these products in cellular DNA have varied widely. Although these two deamination products are often measured by GC-MS analysis, the procedure of acid hydrolysis to release DNA bases for derivatization poses a risk of artifactual deamination of the DNA. In this study, we demonstrated the artifactual formation of these two deamination products during acid hydrolysis and hence developed a method for detecting and measuring 2'-deoxyinosine, the nucleoside of hypoxanthine. Our assay for 2'-deoxyinosine employs nuclease P1 and alkaline phosphatase to achieve release of the nucleosides from DNA, followed by HPLC prepurification with subsequent GC-MS analysis of the nucleosides. This assay detected an increase in the levels of 2'-deoxyinosine in DNA when commercial salmon testis DNA was treated with nitrous acid. We also used it to measure levels in various rat tissues of both normal and endotoxin-treated rats, but could not find increased 2'-deoxyinosine formation in tissues even though *NO production was substantially increased. PMID- 16716896 TI - Silver activates mast cells through reactive oxygen species production and a thiol-sensitive store-independent Ca2+ influx. AB - In genetically susceptible human and/or experimental animals, heavy metals such as mercury, gold, and silver have been shown to highly induce adverse immunological reactions such as allergy and autoimmunity, in which mast cell degranulation is implicated as playing a role. We previously reported that silver activates mast cells and induces Ca2+ influx without stimulating intracellular signaling events required for activation of store-operated Ca2+ channels (SOCs). The purpose of the present study was to elucidate the possible involvement of reactive oxygen species (ROS) in the biological effects of silver. Analysis using oxidant-sensitive fluorescent probes such as dichlorodihydrofluorescein and scopoletin, as well as MCLA-amplified chemiluminescence, showed that silver induced intracellular production and/or extracellular release of ROS. Silver induced mast cell degranulation in a Ca2+ -dependent manner. Unlike IgE antigen, silver-induced Ca2+ influx was not affected by depletion of internal Ca2+ stores. Instead, the metal-induced Ca2+ influx was abolished and reversed by the cell impermeant thiol-reducing agent dithiothreitol, indicating the regulation by oxidation of vicinal thiols on the cell surface. Consistent with this view, Ca2+ influx was blocked by the glutathione peroxidase mimetic ebselen (2-phenyl-1,2 benzisoselenazol-3(2H)-one) and the superoxide dismutase mimetic manganese(III) tetrakis 4-(benzoic acid)porphyrin, but not by exogenously added catalase or superoxide dismutase. These findings indicate that silver evokes the release of ROS and oxidation of thiols critical for the activation of a Ca2+ channel other than SOC. Such a novel ROS-dependent pathway might play a role in mast cell degranulation in metal-induced allergic and autoimmune reactions. PMID- 16716898 TI - Early thymic T cell development in young transgenic mice overexpressing human Cu/Zn superoxide dismutase, a model of Down syndrome. AB - Previous studies have shown that transgenic mice overexpressing Cu/Zn superoxide dismutase, a model of Down syndrome, exhibit premature thymic involution. We have performed a flow cytometry analysis of the developing thymus in these homozygous transgenic mice (hSOD1/hSOD1: Tg-SOD). Longitudinal follow-up analysis from day 3 to day 280 showed an early thymic development in Tg-SOD mice compared with controls. This early thymic development was associated with an increased migration of mature T cells to peripheral lymphoid organs. BrdU labeling showed no difference between Tg-SOD and control mice, confirming that the greater number of peripheral T cells in Tg-SOD mice was not due to extensive proliferation of these cells but rather to a greater pool of emigrant T cells in Tg-SOD. PMID- 16716897 TI - Postischemic hyperoxia reduces hippocampal pyruvate dehydrogenase activity. AB - The pyruvate dehydrogenase complex (PDHC) is a mitochondrial matrix enzyme that catalyzes the oxidative decarboxylation of pyruvate and represents the sole bridge between anaerobic and aerobic cerebral energy metabolism. Previous studies demonstrating loss of PDHC enzyme activity and immunoreactivity during reperfusion after cerebral ischemia suggest that oxidative modifications are involved. This study tested the hypothesis that hyperoxic reperfusion exacerbates loss of PDHC enzyme activity, possibly due to tyrosine nitration or S nitrosation. We used a clinically relevant canine ventricular fibrillation cardiac arrest model in which, after resuscitation and ventilation on either 100% O2 (hyperoxic) or 21-30% O2 (normoxic), animals were sacrificed at 2 h reperfusion and the brains removed for enzyme activity and immunoreactivity measurements. Animals resuscitated under hyperoxic conditions exhibited decreased PDHC activity and elevated 3-nitrotyrosine immunoreactivity in the hippocampus but not the cortex, compared to nonischemic controls. These measures were unchanged in normoxic animals. In vitro exposure of purified PDHC to peroxynitrite resulted in a dose-dependent loss of activity and increased nitrotyrosine immunoreactivity. These results support the hypothesis that oxidative stress contributes to loss of hippocampal PDHC activity during cerebral ischemia and reperfusion and suggest that PDHC is a target of peroxynitrite. PMID- 16716899 TI - Causes of DNA single-strand breaks during reduction of chromate by glutathione in vitro and in cells. AB - Carcinogenic chromates induce DNA single-strand breaks (SSB) that are detectable by conventional alkali-based assays. However, the extent of direct breakage has been uncertain because excision repair and hydrolysis of Cr-DNA adducts at alkaline pH also generate SSB. We examined mechanisms of SSB production during chromate reduction by glutathione (GSH) and assessed the significance of these lesions in cells using genetic approaches. Cr(VI) reduction was biphasic and the formation of SSB occurred exclusively during the slow reaction phase. Catalase or iron chelators completely blocked DNA breakage, as did the use of GSH purified by a modified Chelex procedure. Thus, the direct intermediates of GSH-chromate reactions were unable to cause SSB unless activated by H2O2. SSB repair-deficient XRCC1(-/-) and proficient XRCC1+ EM9 cells had identical survival at doses causing up to 60% clonogenic death and accumulation of 1 mM Cr(VI). However, XRCC1(-/-) cells displayed higher lethality in the more toxic range and the depletion of GSH made them hypersensitive even to moderate doses. Elevation of cellular catalase or GSH levels eliminated survival differences between XRCC1(-/ ) and XRCC1+ cells. In summary, formation of toxic SSB in cells occurs at relatively high chromate doses, requires H2O2, and is suppressed by high GSH concentrations. PMID- 16716900 TI - Absence of CuZn superoxide dismutase leads to elevated oxidative stress and acceleration of age-dependent skeletal muscle atrophy. AB - We describe a novel phenotype in mice lacking the major antioxidant enzyme, CuZn superoxide dismutase (Sod1(-/-) mice), namely a dramatic acceleration of age related loss of skeletal muscle mass. Sod1(-/-) mice are 17 to 20% smaller and have a significantly lower muscle mass than wild-type mice as early as 3 to 4 months of age. Muscle mass in the Sod1(-/-) mice is further reduced with age and by 20 months, the hind-limb muscle mass in Sod1(-/-) mice is nearly 50% lower than in age-matched wild-type mice. Skeletal muscle tissue from young Sod1(-/-) mice has elevated oxidative damage to proteins, lipids, and DNA compared to muscle from young wild-type mice. The reduction in muscle mass and elevated oxidative damage are accompanied by a 40% decrease in voluntary wheel running by 6 months of age and decreased performance on the Rota-rod test at 13 months of age, but are not associated with a decline in overall spontaneous activity. In some of the old Sod1(-/-) mice, the loss in muscle mass is also associated with the presence of tremors and gait disturbances. Thus, the absence of CuZnSOD imposes elevated oxidative stress, loss of muscle mass, and physiological consequences that resemble an acceleration of normal age-related sarcopenia. PMID- 16716901 TI - Oxidative stress induces actin-cytoskeletal and tight-junctional alterations in hepatocytes by a Ca2+ -dependent, PKC-mediated mechanism: protective effect of PKA. AB - Oxidative stress elevates Ca2+ and, presumably, activates Ca2+ -dependent PKCs. We analyzed the participation of Ca2+ -dependent PKCs in actin disorganization and tight-junctional impairment induced by the pro-oxidant tert butylhydroperoxide (tBOOH) in isolated rat hepatocyte couplets. tBOOH (100 microM) augmented radical oxygen species (ROS), as indicated by increased lipid peroxidation (+217%, p < 0.05) and intracellular production of 2',7' dichlorofluorescein (+36%, p < 0.05). Cytosolic Ca2+ and PKCalpha translocation to membrane, an indicator of PKCalpha activation, were also elevated by tBOOH (+100 and +79%, respectively, p < 0.05). tBOOH increased the number of couplets displaying membrane blebs (+278%, p < 0.001) and caused redistribution of F actin. tBOOH induced tight-junctional impairment, as indicated by a reduction in the percentage of couplets retaining presecreted cholyllysylfluorescein in their canalicular vacuoles (-54%, p < 0.001). tBOOH induced redistribution of the tight junctional-associated protein ZO-1. All these events were prevented by the panspecific PKC inhibitors H7 and staurosporine, the Ca2+ -dependent PKC inhibitor Go6976, the intracellular Ca2+ chelator BAPTA/AM, and the PKA activator dibutyryl-cyclic AMP. Furthermore, PKC inhibition and PKA activation not only prevented but also fully reversed tBOOH-induced blebbing. Conversely, tBOOH induced ROS formation and Ca2+ elevation remained unchanged. We conclude that ROS induce hepatocellular actin-cytoskeleton rearrangement and tight-junctional impairment by a PKC-mediated, Ca2+ -dependent mechanism, which is counteracted by PKA. PMID- 16716902 TI - Chain scission of hyaluronan by carbonate and dichloride radical anions: potential reactive oxidative species in inflammation? AB - The reactions of the carbonate and dichloride radical anions, CO3- and Cl2-, with the extracellular matrix glycosaminoglycan hyaluronan (HA) have been studied using the kinetic technique of pulse radiolysis and also by steady-state irradiation combined with gel permeation chromatography/multiangle laser light scattering(gpc/MALLS) to measure the rates of reaction with HA and the yield of HA chain scission, respectively. For comparison, the same measurements were made for the reactions of the free radicals *OH, Br2*-, and N3*. The carbonate and dichloride radical anions were found to react relatively quickly with HA (7.0 x 10(5) and 6.9 x 10(6) dm3 mol(-1) s(-1), respectively) although they are much less reactive than the hydroxyl radical, *OH. Significant yields (20 and 38%, respectively) of chain scission of HA by these radical anions were also determined from the gpc/MALLS experiments, providing some support for their potential participation in the depolymerization of HA in vivo. These results are compared with data obtained for the other free radicals (hydroxyl, azide radicals, and dibromide radical anions) investigated in this study in order to gain an insight into their mechanism of reaction with HA. Earlier chain scission yields of HA by hydroxyl radicals determined by the authors have also been revised using the gpc/MALLS technique employed in the current study. The yields of 52% (absence of air) and 44% (in air) are much lower than the previous values. In the current study, the effect of oxygen on the yields of HA chain breaks is discussed in terms of the reactivity of HA peroxyl radicals in the presence of superoxide radical anions. The relevance of the results of this study to mechanisms of inflammation is discussed. PMID- 16716903 TI - Reduction in molecular synthesis or enzyme activity of superoxide dismutases and catalase contributes to oxidative stress and neurogenic hypertension in spontaneously hypertensive rats. AB - A balance between production and elimination of reactive oxygen species such as superoxide anion (O2*-) and hydrogen peroxide (H2O2) tightly regulates the homeostasis of cellular oxidative stress, which contributes to a variety of cardiovascular diseases, including hypertension. The present study assessed the hypothesis that O2*- or H2O2 levels augmented by the reduced molecular synthesis or enzyme activity of superoxide dismutase (SOD), catalase (CAT), or glutathione peroxidase (GPx) in the rostral ventrolateral medulla (RVLM), where sympathetic premotor neurons that generate tonic vasomotor tone are located, contribute to the pathogenesis of hypertension. We found that copper/zinc SOD (SOD1), manganese SOD (SOD2), or CAT, but not GPx, mRNA or protein expression and enzyme activity in the RVLM of spontaneously hypertensive rats (SHR) were significantly lower than those in normotensive Wistar-Kyoto (WKY) rats, along with a significantly higher level of O2*- or H2O2. A causative relationship between these biochemical correlates of oxidative stress and neurogenic hypertension was established when gene transfer by microinjection of adenovirus encoding SOD1, SOD2, or CAT into the bilateral RVLM promoted a long-lasting reduction in arterial pressure in SHR, but not WKY rats, accompanied by an enhanced SOD1, SOD2, or CAT protein expression or enzyme activity and reduced O2*- or H2O2 level in the RVLM. These results together suggest that downregulation of gene expression and enzyme activity of the antioxidant SOD1, SOD2, or CAT may underlie the augmented levels of O2*- and H2O2 in the RVLM, leading to oxidative stress and hypertension in SHR. PMID- 16716904 TI - In vivo detection of free radicals induced by diethylnitrosamine in rat liver tissue. AB - Diethylnitrosamine (DEN) is a well-known carcinogenic substance that requires microsomal activation before it can react with DNA to cause mutations and cancer. The aim of this study was to use in vivo spin trapping and spin probe techniques to investigate whether free radicals are generated in rat liver tissue during DEN activation. We used alpha-phenyl-n-tert-butylnitrone (PBN) as the spin trapping agent, which was delivered through an intraperitoneal injection before DEN administration. One hour after DEN administration, multicomponent PBN adducts in the bile were detected, and the intensities were diminished by the cytochrome P450 inhibitor SKF-525A. A computer simulation of the ESR signals revealed the presence of a lipid-derived radical. Using the in vivo spin probe/ESR technique, the signal decay rate of methoxycarbonyl-PROXYL was significantly increased in the DEN-treated group compared with the rate in the vehicle group. The enhanced signal decay rate was restored with PBN and/or SKF-525A pretreatment. These results suggested that lipid-derived free radicals were generated in the liver within 1 h after DEN administration. PMID- 16716905 TI - Rapid intracellular acidification and cell death by H2O2 and alloxan in pancreatic beta cells. AB - Pancreatic beta-cell death induced by oxidative stress plays an important role in the pathogenesis of diabetes mellitus. We studied the relation between rapid intracellular acidification and cell death of pancreatic beta-cell line NIT-1 cells exposed to H2O2 or alloxan. Intracellular pH was measured by a pH-sensitive dye, and cell damage by double staining with Annexin-V and propidium iodide using flow cytometry. H2O2 and alloxan caused a rapid fall in intracellular pH and suppressed Na+/H+ exchanger activity in the NH4Cl prepulse method. H2O2 induced necrotic cell death, which shifted to apoptotic cell death when initial acidification was prevented by pH clamping to 7.4 using nigericin (unclamped cells vs clamped cells, necrosis 43.8 +/- 5.8% vs 21.1 +/- 10.6%, P < 0.05; apoptosis 8.0 +/- 1.9% vs 44.5 +/- 5.0%, P < 0.01). pH-clamped cells showed enhanced caspase 3 activity and proapoptotic Bax expression. On the other hand, NIT-1 cells were resistant to alloxan toxicity, but treatment with alloxan and nigericin strikingly enhanced the cytotoxicity. Antioxidants partly prevented cell death, although intracellular pH remained similarly acidic. The rapid intracellular acidification was not the cause of cell death but a significant determinant of the mode of death of H2O2 -treated beta cells, whereas no link between cell death and acidification was demonstrated in alloxan toxicity. PMID- 16716906 TI - Endothelial cell superoxide anion radical generation is not dependent on endothelial nitric oxide synthase-serine 1179 phosphorylation and endothelial nitric oxide synthase dimer/monomer distribution. AB - Tetrahydrobiopterin (BH4) and heat shock protein 90 (hsp90) have been anticipated to regulate endothelial nitric oxide synthase (eNOS)-dependent superoxide anion radical (O2*-) generation in endothelial cells. It is not known, however, whether hsp90 and BH4 increase O2*- in a synergistic manner, or whether this increase is a consequence of downstream changes in eNOS phosphorylation on serine 1179 (eNOS S1179) and changes in dimer/monomer distribution. Here O2*- production from purified BH4 -free eNOS and eNOS:hsp90 complexes determined by spin-trapping methodology showed that hsp90 neither inhibits O2*- nor alters the requirement of BH4 to inhibit radical release from eNOS. In endothelial cells, O2*- detection with the novel high-performance liquid chromatography assay of 2-hydroxyethidium showed that inhibition of hsp90 did not increase O2*-, while a significant increase in O2*- was detected in BH4 -depleted cells. Radicicol, a hsp90 inhibitor, disrupted eNOS:hsp90 association, decreased eNOS-S1179, but increased biopterin production in a dose-dependent fashion. These changes were followed by an increase in eNOS activity, demonstrating that high biopterin levels offset inhibition of eNOS phosphorylation and diminished interaction with hsp90. In contrast, depletion of biopterin did not affect hsp90 levels or interaction with eNOS or eNOS dimer/monomer ratio in bovine aorta endothelial cells (BAECs). We conclude that low BH4 but not inhibition of hsp90 increases O2*- in BAECs by mechanism(s) that unlikely involve phosphorylation to eNOS-S1179 or eNOS monomerization. PMID- 16716907 TI - A probabilistic model of gaze imitation and shared attention. AB - An important component of language acquisition and cognitive learning is gaze imitation. Infants as young as one year of age can follow the gaze of an adult to determine the object the adult is focusing on. The ability to follow gaze is a precursor to shared attention, wherein two or more agents simultaneously focus their attention on a single object in the environment. Shared attention is a necessary skill for many complex, natural forms of learning, including learning based on imitation. This paper presents a probabilistic model of gaze imitation and shared attention that is inspired by Meltzoff and Moore's AIM model for imitation in infants. Our model combines a probabilistic algorithm for estimating gaze vectors with bottom-up saliency maps of visual scenes to produce maximum a posteriori (MAP) estimates of objects being looked at by an observed instructor. We test our model using a robotic system involving a pan-tilt camera head and show that combining saliency maps with gaze estimates leads to greater accuracy than using gaze alone. We additionally show that the system can learn instructor specific probability distributions over objects, leading to increasing gaze accuracy over successive interactions with the instructor. Our results provide further support for probabilistic models of imitation and suggest new ways of implementing robotic systems that can interact with humans over an extended period of time. PMID- 16716908 TI - Antifungal effects of the volatile oils from Allium plants against Trichophyton species and synergism of the oils with ketoconazole. AB - In an attempt to develop stable and safe antifungal agents from natural products (daily foodstuffs in particular), the activities of essential oils from Allium sativum for. pekinense, A. cepa, and A. fistulosum against three Trichophyton species responsible for severe mycoses in humans were investigated and compared with activity of allicin in this study. The fungistatic activities of Allium oils were evaluated by the broth dilution method and disk diffusion assay. The combined effects of Allium oils with ketoconazole were tested by the checkerboard titer test. Among the tested oils, A. sativum for. pekinense oil exhibited the strongest inhibition of growth of T. rubrum, T. erinacei, and T. soudanense with MICs (minimum inhibiting concentrations) of 64microg/ml, while the activities of A. cepa and A. fistulosum were relatively mild. The inhibiting activities of the oils on Sabouraud agar plates were dose dependent against Trichophyton species. Additionally, these oils showed significant synergistic antifungal activity when combined with ketoconazole in the checkerboard titer test and disk diffusion test. PMID- 16716909 TI - Screening of novel nuclear receptor agonists by a convenient reporter gene assay system using green fluorescent protein derivatives. AB - Nuclear receptors represent a very good family of protein targets for the prevention and treatment of diverse diseases. In this study, we screened natural compounds and their derivatives, and discovered ligands for the retinoic acid receptors (RARs) and the farnesoid X receptor (FXR). In the reporter assay systems of nuclear receptors presented here, two fluorescent proteins, enhanced yellow fluorescent protein (EYFP) and enhanced cyan fluorescent protein (ECFP), were used for detection of a ligand-based induction and as an internal control, respectively. By optimizing the conditions (e.g., of hormone response elements and promoter genes for reporter plasmids), we established a battery of assay systems for ligands of RARs, retinoid X receptor (RXR) and FXR. The screening using the reporter assay system can be carried out without the addition of co factors or substrates. As a result of screening of more than 140 compounds, several compounds were detected which activate RARs and/or FXR. Caffeic acid phenylethyl ester (CAPE), known as a component of propolis from honeybee hives, and other derivatives of caffeic acid up-regulated the expression of reporter gene for RARs. Grifolin and ginkgolic acids, which are non-steroidal skeleton compounds purified from mushroom or ginkgo leaves, up-regulated the expression of the reporter gene for FXR. PMID- 16716910 TI - Protective effects of a standard extract of Mangifera indica L. (VIMANG) against mouse ear edemas and its inhibition of eicosanoid production in J774 murine macrophages. AB - A standard aqueous extract of Mangifera indica L., used in Cuba as antioxidant under the brand name VIMANG, was tested in vivo for its anti-inflammatory activity, using commonly accepted assays. The standard extract of M. indica, administered orally (50-200mg/kg body wt.), reduced ear edema induced by arachidonic acid (AA) and phorbol myristate acetate (PMA) in mice. In the PMA model, M. indica extract also reduced myeloperoxidase (MPO) activity. In vitro studies were performed using macrophage cell line J774 stimulated with pro inflammatory stimuli lipopolysaccharide-interferon gamma (LPS-IFNgamma) or calcium ionophore A23187 to determine prostaglandin PGE(2) or leukotriene LTB(4) release, respectively. The extract inhibited the induction of PGE(2) and LTB(4) with IC(50) values of 21.7 and 26.0microg/ml, respectively. Mangiferin (a glucosylxanthone isolated from the extract) also inhibited these AA metabolites (PGE(2), IC(50) value=17.2microg/ml and LTB(4), IC(50) value=2.1microg/ml). These results represent an important contribution to the elucidation of the mechanism involved in the anti-inflammatory and anti-nociceptive effects reported for the standard extract of M. indica VIMANG. PMID- 16716911 TI - In vitro effects of genistein and resveratrol on the production of interferon gamma (IFNgamma) and interleukin-10 (IL-10) by stimulated murine splenocytes. AB - Phytoestrogens are a group of plant-derived biologically active substances with a chemical structure that resembles that of 17beta-estradiol (E2). As the presence of estrogen receptors (ER) has been identified in several immune cells, phytoestrogens may also have a great impact on the immune system. The aim of our study was to determine the in vitro effects of genistein and resveratrol on the production of interferon-gamma (IFNgamma) and interleukin-10 (IL-10) by stimulated murine splenocytes and compare them with the effects of natural E2. Phorbol 12-miristate 13-acetate (PMA) together with ionomycin was used to stimulate the cells. E2 and genistein did not show any significant effects on the stimulated production of IFNgamma. Resveratrol had a mild inhibitory effect on IFNgamma production at the concentration of 10(-7)M; however, this difference did not reach statistical significance (p>0.05). IL-10 levels in the splenocytes culture supernatants were found to be increased in the presence of E2, genistein and resveratrol; however, these effects were also not statistically significant. To determine whether the exposure to our studied phytoestrogens induced a shift in the T-helper 1/T-helper 2 (Th1/Th2) balance, we calculated the ratio between the production of IFNgamma, the prototypic Th1 cytokine, and the production of IL 10, the prototypic Th2 cytokine, at different concentrations of our tested substances. Genistein at the concentrations of 10(-6) and 10(-7)M and resveratrol at the concentrations of 10(-6)M decreased significantly the IFNgamma/IL-10 ratio. This decrease was comparable to that of E2 at the concentrations of 10( 7)M. From our in vitro experiments we conclude that genistein and resveratrol, similarly to E2, by decreasing the IFNgamma/IL10 ratio may shift the Th1/Th2 balance towards the Th2 response. PMID- 16716912 TI - Binding studies of an arabinogalactan-protein from Echinacea purpurea to leucocytes. AB - Flow cytometric investigations show binding of an isolated arabinogalactan protein (AGP) from pressed juice of the aerial parts of Echinacea purpurea to the cell surface of human leucocytes. AGP demonstrates binding to lymphocytes, monocytes and granulocytes of different donors (n=8). Competition assays with two antibodies, directed against CD4 and CD8, revealed no interaction of AGP with these receptors, leading to the conclusion that binding of AGP to leucocytes is mediated via other structures. PMID- 16716913 TI - Hypoglycemic activity of polysaccharide, with antioxidation, isolated from cultured Cordyceps mycelia. AB - Cordyceps sinensis, a well-known traditional Chinese medicine, possesses anti tumor, immunostimulant and antioxidant activities; however, the identities of active components have not been determined. In our previous study using antioxidant activity-guided fractionation [Li et al., 2003. A polysaccharide isolated from Cordyceps sinensis, a traditional Chinese medicine, protects PC12 cells against hydrogen peroxide-induced injury. Life Sci. 73, 2503-2513], a polysaccharide of molecular weight approximately 210kDa was isolated from cultured Cordyceps mycelia by ion-exchange and sizing chromatography. The isolated polysaccharide, named CSP-1, which has strong anti-oxidation activity, contains glucose, mannose and galactose in the ratio of 1:0.6:0.75. In the present study, we demonstrated the hypoglycemic effect of CSP-1 on normal and alloxan-diabetic mice and streptozotocin (STZ)-diabetic rats. The basal glucose level did not differ significantly among the normal mice. CSP-1 (at 200 and 400mg/kg body wt./day for 7 days, p.o.), however, significantly reduced the blood glucose level by 12.0+/-3.2% and 22.5+/-4.7% in normal mice, respectively (p<0.05). When administered at a dose of higher than 200mg/kg body wt. daily for 7 days, CSP-1 produced a significant drop in blood glucose level in both STZ induced diabetic rats and alloxan-induced diabetic mice. The serum insulin levels in diabetic animals were also increased by administration of CSP-1 (p<0.05). CSP 1 with hypoglycemic properties increased circulating insulin level in diabetic animals, which suggests that CSP-1 may stimulate pancreatic release of insulin and/or reduce insulin metabolism. PMID- 16716914 TI - In vitro glucose uptake activity of Aegles marmelos and Syzygium cumini by activation of Glut-4, PI3 kinase and PPARgamma in L6 myotubes. AB - The purpose of the present study is to investigate the effect of methanolic extracts of Aegles marmelos and Syzygium cumini on a battery of targets glucose transporter (Glut-4), peroxisome proliferator activator receptor gamma (PPARgamma) and phosphatidylinositol 3' kinase (PI3 kinase) involved in glucose transport. A. marmelos and S. cumini are anti-diabetic medicinal plants being used in Indian traditional medicine. Different solvent extracts extracted sequentially were analysed for glucose uptake activity at each step and methanol extracts were found to be significantly active at 100ng/ml dose comparable with insulin and rosiglitazone. Elevation of Glut-4, PPARgamma and PI3 kinase by A. marmelos and S. cumini in association with glucose transport supported the up regulation of glucose uptake. The inhibitory effect of cycloheximide on A. marmelos- and S. cumini-mediated glucose uptake suggested that new protein synthesis is required for the elevated glucose transport. Current observation concludes that methanolic extracts of A. marmelos and S. cumini activate glucose transport in a PI3 kinase-dependent fashion. PMID- 16716915 TI - Vascular effects of 7-epiclusianone, a prenylated benzophenone from Rheedia gardneriana, on the rat aorta. AB - The vascular effects of 7-epiclusianone on the rat aorta were investigated. In the rat aortic rings with functional endothelia, 7-epiclusianone up to 10microM induced a concentration-dependent vasodilatation of the sustained contractions induced by phenylephrine (0.3microM). At concentrations higher than 10microM, 7 epiclusianone induced a concentration-dependent contraction in the aortic rings. The vasodilator effect of 7-epiclusianone was drastically decreased with L-NAME (100microM) as well as in endothelium-denuded aortic rings. Moreover, indomethacin (10microM) induced a significant shift to the left in the vasodilator but did not modify the vasoconstrictor effect of 7-epiclusianone. In arteries without pre-contraction, 7-epiclusianone (3-100microM) induced concentration-dependent contraction only in endothelium-intact and in the presence of L-NAME (100microM). This effect was inhibited by indomethacin (10microM) and ZM230487 (1microM), selective inhibitors of cyclooxygenase and of 5-lipoxygenase, respectively. We can conclude that at low concentrations 7 epiclusianone induces an endothelium-dependent vasodilator effect in rat aortic rings. At higher concentrations and in conditions where NO synthase was inhibited, 7-epiclusianone induces a vasocontractile effect. Nitric oxide seems to participate in the vasodilatation, while endothelial cyclooxygenase- and 5 lipoxygenase-derived products play a role in the vasoconstrictor effect. PMID- 16716916 TI - The role of adrenal corticosteroids in the anti-inflammatory effect of the whole extract of Harpagophytum procumbens in rats. AB - This study was carried out to evaluate whether the anti-inflammatory response in rats to the whole extract of Harpagophytum procumbens is a consequence of adrenal corticosteroid release. Carrageenan-induced inflammatory responses in the hindpaws were evaluated in control, sham-operated and adrenalectomized rats. The extract was administered orally (by gavage) or intraperitoneally, 30min prior to injury stimulus. Blood samples were then collected, and the number of circulating leukocytes was estimated. Pretreatment with the whole extract of H. procumbens reduced the intensity of inflammatory response in normal, sham-operated and adrenalectomized animals. When administered orally, the extract was ineffective. The reduced number of circulating leukocytes observed following intraperitoneal injection of the extract characterized adrenal hyperactivity. The inhibitory effect of the whole extract of H. procumbens on acute inflammatory response in the rat, when administered intraperitoneally, does not depend on the release of adrenal corticosteroids. PMID- 16716917 TI - Knipholone, a selective inhibitor of leukotriene metabolism. AB - Inhibition of leukotriene formation is one of the approaches to the treatment of asthma and other inflammatory diseases. We have investigated knipholone, isolated from the roots of Kniphofia foliosa, Hochst (Asphodelaceae), for inhibition of leukotriene biosynthesis in an ex vivo bioassay using activated human neutrophile granulocytes. Moreover, activities on 12-lipoxygenase from human platelets and cycloxygenase (COX)-1 and -2 from sheep cotyledons and seminal vesicles, respectively, have been evaluated. Knipholone was found to be a selective inhibitor of leukotriene metabolism in a human blood assay with an IC(50) value of 4.2microM. However, at a concentration of 10microg/ml, the compound showed weak inhibition of 12(S)-HETE production in human platelets and at a concentration of 50microM it produced no inhibition of COX-1 and -2. In our attempt to explain the mechanism of inhibition, we examined the antioxidant activity of knipholone using various in vitro assay systems including free radical scavenging, non-enzymatic lipid peroxidation, and metal chelation. Knipholone was found to be a weak dose-independent free radical scavenger and lipid peroxidation inhibitor, but not a metal chelator. Therefore, the leukotriene biosynthesis inhibitory effect of knipholone was evident by its ability either to inhibit the 5-lipoxygenase activating protein (FLAP) or as a competitive (non-redox) inhibitor of the enzyme. Cytotoxicity results also provided evidence that knipholone exhibits less toxicity for a mammalian host cell. PMID- 16716918 TI - Video-assisted thoracoscopic lobectomy in infants. AB - OBJECTIVE: Congenital lung malformations are often discovered on routine prenatal sonography or postnatal imaging. Lesions such as congenital cystic adenomatoid malformation or pulmonary sequestration may be asymptomatic at birth, and their management is controversial. Thoracoscopy in children has been mainly used for lung biopsy and for the treatment of empyema and recurrent pneumothorax. Very few reports of more technically demanding procedures, such as lobectomy, are currently available. This report evaluates the safety and efficacy of video assisted thoracoscopic (VATS) lobectomy in infants and small children with asymptomatic prenatally diagnosed lung lesions. METHODS: During 2004, six patients underwent VATS lobectomy without a mini-thoracotomy. Mean age was 10 months (range, 6-19 months). Preoperative diagnosis included congenital cystic adenomatoid malformation (n = 5) and an extralobar pulmonary sequestration. All patients were asymptomatic and surgery was performed electively. Three or four 3 5mm ports were used. Single lung ventilation and controlled low pressure pneumothorax were used in every case. A bipolar sealing device was the preferred mode of vessel ligation and bronchi were closed with interrupted sutures. A chest tube was left in all cases. RESULTS: All the procedures were completed thoracoscopically. Operating times ranged from 70 to 215 min (mean, 130 min). There were five lower lobe and one middle lobe resections. There were no intraoperative complications and chest tubes were left in place 1-4 days. Two patients showed postoperative hemothorax that stopped spontaneously. Hospital stay ranged from 4 to 9 days (mean, 7 days). CONCLUSIONS: VATS lobectomy in small infants is a feasible and safe technique. Decreased postoperative pain, a shorter hospital stay, and a better cosmetic result are definite advantages of this minimally invasive procedure. Long-term morbidity due to a major thoracotomy incision is avoided. PMID- 16716919 TI - Immunogenic and allergenic potentials of natural and recombinant innocuous proteins. AB - A new aspect of protein immunogenic and allergenic properties has become important recently, when there is a higher chance that our immune system will be exposed to novel protein antigens and/or familiar protein antigens with an unprecedented high frequency and large amount. These proteins are innocuous, nontoxic, and noninvasive by themselves, and include various natural proteins from the environment and recombinant proteins from industry. The technical term allergenic has been used for such proteins and their abilities to induce specific IgE production and to cross-link IgE/Fc epsilonRI on the surface of mast cells and basophiles have been recognized. As for the environmental proteins, some physicochemical properties (solubility, stability, and permeability across a mucosal epithelium) of the proteins indirectly play important roles in their allergenic potential because they do not originate from invasive pathogens as vehicles. Indeed, several lines of experimental evidences have been accumulated indicating that all proteins are absorbed across mucosal epithelia by transcellular transport and/or through interstitial spaces among the epithelial cells but not at equal levels. Some animal models have been established for natural sensitization to some allergenic proteins by feeding or intragastric administration without an adjuvant and, in a few cases, some symptoms resembling human allergy and even anaphylaxis have been induced by oral challenge with the proteins. Sometimes, even to self-proteins, the immunogenic or allergenic potential is given by post-translational modifications and possibly by unknown structural/conformational alterations, when they are exogenous self-proteins, such as recombinant human proteins for drug use. Despite the accumulation of knowledge and the progress in analytical technology on protein allergenicity, it is still crucial to predict the allergenic potential of novel and unused proteins. However, some animal models are applicable for assessing the relative allergenic potential of processed proteins in comparison with that of native proteins in preclinical studies. PMID- 16716920 TI - N-linked glycan structures of a mouse monoclonal antibody produced from tobacco BY2 suspension-cultured cells. AB - cDNA encoding H- and L-chains from a mouse monoclonal antibody was introduced into tobacco BY2 cells, and the resulting sugar chain structures of plant produced antibodies were analyzed by a combination of HPLC, exoglucosidase digestion and matrix-assisted laser desorption/ionization time-of-flight (MALDI TOF) mass spectrometry. The glycan structures determined were Man(5-6)GlcNAc2 (22.3%), GlcNAcMan5GlcNAc2 (3.1%), GlcNAcMan3FucXylGlcNAc2 (24.4%), GlcNAcMan3XylGlcNAc2 (17.8%), Man3FucXylGlcNAc2 (24.3%), and Man3XylGlcNAc2 (8.1%). The major glycan structures of the antibodies produced by transgenic suspension-cultured cells contain typical plant bisecting beta(1,2)-xylose and alpha(1,3)-fucose residues, suggesting the posttranslational modification of a recombinant antibody in the late Golgi apparatus. PMID- 16716921 TI - Bioconversion of D-psicose to D-tagatose and D-talitol by Mucoraceae fungi. AB - Rhizopus oryzae MYA-2483, which cannot utilize D-psicose as a sole source of carbon, converted D-psicose to two other compounds. These compounds were identified by NMR and IR as D-tagatose and D-talitol. In this study, we describe for the first time the bioconversion of D-psicose to D-tagatose. Various strains of Mucoraceae fungi, to which R. oryzae MYA-2483 belongs, exhibited conversion activity similar to that of R. oryzae MYA-2483. There is the possibility that a considerable number of fungi belonging to Mucoraceae possess such D-psicose conversion activity. PMID- 16716922 TI - Effect of salt concentration on intracellular accumulation of lipids and triacylglyceride in marine microalgae Dunaliella cells. AB - In order to get the high liquefaction yield from marine algae cell mass to fuel oil, the effect of salt stress on the accumulation of lipids and triacylglyceride in Dunaliella cells was investigated. Although initial NaCl concentration higher than 1.5 M markedly inhibited cell growth, increase of initial NaCl concentration from 0.5 (equal to sea water) to 1.0 M resulted in a higher intracellular lipid content (67%) in comparison with 60% for the salt concentration of 0.5 M. Addition of 0.5 or 1.0 M NaCl at mid-log phase or the end of log phase during cultivation with initial NaCl concentration of 1.0 M further increased the lipid content (70%). PMID- 16716923 TI - Effects of methanol feeding methods on chimeric alpha-amylase expression in continuous culture of Pichia pastoris. AB - The effects of two types of methanol feeding methods in a continuous culture of Pichia system on the cell growth and recombinant protein expression were studied using chimeric alpha-amylase as a model protein. With the feeding of methanol by a DO-stat method, the alpha-amylase concentration in the fermentation broth increased with decreasing dilution rate and reached 173 mg/l at a dilution rate of 0.013 h(-1), at which the maximum volumetric productivity of alpha-amylase was obtained. Although almost the same productivity was attained at 0.04 h(-1) with continuous methanol feeding, the alpha-amylase concentration was one third that compared with feeding by the DO-stat method, that is, 55 mg/l. Furthermore, at this dilution rate, the medium volume needed per unit time was three times that required when DO-stat was used. Therefore, continuous culture with methanol feeding by the DO-stat method may be a promising method for the production of recombinant proteins on an industrial scale by Pichia pastoris. PMID- 16716924 TI - Effects of water-soluble carbohydrate content on silage fermentation of wheat straw. AB - To determine a suitable initial water-soluble carbohydrate (WSC) content to make wheat straw natural fermentation successful and to study fermentation characteristics, glucose was used to adjust the initial WSC content to 1.4%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, and 10.0% dry matter (DM) in the wheat straw. At 30 d of fermentation, there were three pHs: when the initial WSC content was 1.4%, the pH was 5.5; when the initial WSC contents were 4.0%, 5.0% and 6.0%, the pHs were near 5.1; and when the initial WSC contents were 7.0%, 8.0%, 9.0% and 10.0%, the pHs were near 4.0. The pattern of changes in WSC content during the fermentation was similar to that in pH. At 30 d of fermentation, there existed a dividing line in WSC remnants between the initial 6.0% WSC treatment and the initial 7.0% WSC treatment. When the initial WSC content was more than 7.0%, the remaining WSC content was more than 23.7 g/kg DM. When the initial WSC content was less than 6.0%, the remaining WSC content was less than 13.6 g/kg DM. Particularly for the 1.4% WSC treatment, the remaining WSC content was 2.1 g/kg DM. The results of the microbiological enumeration showed that with the increase in initial WSC content, the numbers of lactic acid bacteria (LAB) and other bacteria generally decreased. Denaturing gradient gel electrophoresis (DGGE) results showed that when the initial WSC content was beyond 7.0%, the LAB of the fermentation system were detected. PMID- 16716925 TI - Morphology of spheroidal hepatocytes within injectable, biodegradable, and thermosensitive poly(organophosphazene) hydrogel as cell delivery vehicle. AB - An aggregate of specific cells is often regarded as a better form of single-cell in artificial organs and mammalian cell bioreactors for improved cell-specific functions. In this study, freshly harvested primary rat hepatocytes, cultivated as spheroids and entrapped in a synthetic thermoreversible extracellular matrix, were examined for differentiation morphology and enhanced liver-specific functions and compared with a control set (single-cell hepatocytes). A copolymer of poly(organophsophazene) hydrogel, an injectable, biodegradable, and thermosensitive matrix, was used to entrap hepatocytes as spheroids or single cells. In a 28-d culture period, the spheroidal hepatocytes maintained a higher viability and produced albumin and urea at constant rates, whereas the single hepatocytes showed a reduction in the level of albumin secretion in the hydrogel. The culturing of spheroidal hepatocytes is a potentially useful three-dimensional cell system for application in bioartificial liver devices and bioreactors. PMID- 16716926 TI - Gene expression property of high-density three-dimensional tissue of HepG2 cells formed in radial-flow bioreactor. AB - In our previous study, we examined three-dimensional culture using 5-ml radial flow bioreactor (RFB) and showed that genes encoding cell cycle related proteins were suppressed in a stable phase. In this study, we analyzed the gene expression profiles of RFB-cultivated HepG2 cells and found that vascular endothelial growth factor (VEGF) production was strongly induced in the stable phase compared with the growth phase or static two-dimensional culture. When human umbilical vein endothelial cells (HUVECs) were grown under the conditioned medium of the stable phase, it was found that the formation of new blood vessels was induced in the angiogenesis model. DNA microarray analysis showed that the expression levels of both genes related to cell cycle arrest and which are known as tumor markers have increased in the stable phase. This result suggests that HepG2 cells in the stable phase maintain an active tumor phenotype. In addition, the expression of genes induced in the hypoxic condition was also induced in the stable phase. When the culture was carried out under a higher dissolved oxygen (DO) concentration, VEGF production did not decrease significantly and the new blood-vessel-forming ability of the conditioned medium was not suppressed. This suggests that the induction of VEGF production in a stable phase is not affected by DO during the tested level. These results suggest that the RFB cell culture system may be used to assess tumor progression mechanism under three-dimensional condition in vitro. PMID- 16716927 TI - Production of Fab fragment corresponding to surface protein antigen of Streptococcus mutans serotype c-derived peptide by Escherichia coli and cultured tobacco cells. AB - The cDNA of a mouse Fab fragment was cloned from a hybridoma cell line that produces a mouse monoclonal antibody, KH5, that reacts with the peptide fragment of the surface protein antigen of Streptococcus mutans serotype c (PAc). After transfection with cDNA, recombinant Fab fragments were produced by Escherichia coli (T15 Fab) and cultured tobacco cells (X253 and X262 Fabs). The antipeptide activities of T15 and X253 were similar to that of KH5. X253 was secreted into the culture media, which had a specific affinity for the PAc peptide. PMID- 16716928 TI - Fate of transforming bacterial genome following incorporation into competent cells of Bacillus subtilis: a continuous length of incorporated DNA. AB - In contrast to the conventional transformation of Bacillus subtilis using purified DNA, those using DNA in lysed protoplasts have a high transformation efficiency and enable whole-genome transfer into competent B. subtilis [Akamatsu, T. and Taguchi, H., Biosci. Biotechnol. Biochem., 65, 823-829 (2001)]. Here, we examined the length of incorporated continuous DNA by analyzing the cotransfer ratio with selected and unselected markers, on the basis of a new experimental design. The cotransfer ratio of a selected marker with an unselected marker on the opposite side of the genetic map of the B. subtilis chromosome was about 5.6% and could be interpreted as congression (double transformation) ratio. In the wild-type strain, the cotransfer ratio of cysA (113 kb position on 4215 kb of B. subtilis chromosome) with metC (1384 kb) and leuB (2891 kb) was 0.77%, twice the value (5.6% x 5.6%=0.31%) calculated from the congression ratio. Moreover, in a genetic background, the cotransfer ratios of metC with cysA and leuB, and metC with cysA and arg1 (3012 kb) were 2.7% and 7.2%, respectively. These results strongly suggest that the length of continuous DNA incorporated into B. subtilis is most probably greater than 1271 kb. When the DNA from the protoplast lysate was fragmented by mixing, the cotransfer ratios of arg1 with metC, and arg1 with metC and trpC (2374 kb) were 2.8% and 0.16%, respectively. A high cotransfer ratio (2.7-7.2%) could not, therefore, be obtained using the fragmented DNA. Based on these observations, we propose a working hypothesis on the mechanism of the transformation of competent B. subtilis by DNA in protoplast lysates (LP transformation). PMID- 16716929 TI - Cloning and characterization of nif structural and regulatory genes in the purple sulfur bacterium, Halorhodospira halophila. AB - Halorhodospira halophila is a halophilic photosynthetic bacterium classified as a purple sulfur bacterium. We found that H. halophila generates hydrogen gas during photoautotrophic growth as a byproduct of a nitrogenase reaction. In order to consider the applied possibilities of this photobiological hydrogen generation, we cloned and characterized the structural and regulatory genes encoding the nitrogenase, nifH, nifD and nifA, from H. halophila. This is the first description of the nif genes for a purple sulfur bacterium. The amino-acid sequences of NifH and NifD indicated that these proteins are an Fe protein and a part of a MoFe protein, respectively. The important residues are conserved completely. The sequence upstream from the nifH region and sequence similarities of nifH and nifD with those of the other organisms suggest that the regulatory system might be a NifL-NifA system; however, H. halophila lacks nifL. The amino acid sequence of H. halophila NifA is closer to that of the NifA of the NifL-NifA system than to that of NifA without NifL. H. halophila NifA does not conserve either the residue that interacts with NifL or the important residues involved in NifL-independent regulation. These results suggest the existence of yet another regulatory system, and that the development of functional systems and their molecular counterparts are not necessarily correlated throughout evolution. All of these Nif proteins of H. halophila possess an excess of acidic residues, which acts as a salt-resistant mechanism. PMID- 16716930 TI - Microbial community in methanogenic packed-bed reactor successfully operating at short hydraulic retention time. AB - The microbial community in a thermophilic anaerobic packed-bed reactor, which had been successfully operated to convert acetic and butyric acids to methane at a short hydraulic retention time (from 24 h to 1.9 h), was investigated. Archaea closely related to known methanogens were detected by 16S rRNA gene analyses of the effluents, together with diverse types of unidentified bacteria. PMID- 16716931 TI - Robustness of cascade pH and dissolved oxygen control in symbiotic nisin production process system of Lactococcus lactis and Kluyveromyces marxianus. AB - In symbiotic processes, different organisms coexist stably and interact by sharing the same metabolites and environmental conditions. The robustness of a symbiotic nisin production process system composed of the lactic acid bacterium Lactococcus lactis subsp. lactis (ATCC11454) and dairy yeast Kluyveromyces marxianus (MS1) was studied. It was found that this symbiotic process system was robust to the initial disturbance in the inoculum sizes of both microorganisms and pH. PMID- 16716932 TI - Quantitative expression analysis of inulinase gene cluster of Penicillium sp. strain TN-88. AB - The filamentous fungus Penicillium sp. strain TN-88 carries the endoinulinase gene inuC and the exoinulinase gene inuD that are linked head-to-head on the genome and divergently transcribed from an 859-bp intergenic region [Moriyama et al., Biosci. Biotechnol. Biochem., 66, 1887-1896 (2002)]. Quantitative real-time PCR amplification revealed that the transcription levels of the inuC and inuD genes increased 42- and 3260-fold in inulin-grown mycelia, respectively. Sucrose as well as fructose did not induce the expression of the inuC or inuD gene at all. The levels of inuC and inuD transcripts in mycelia grown on the glucose/inulin mixture were both below their basal levels in glucose-grown mycelia. Thus, glucose exerts a strong carbon catabolite repression on the expression of the two genes. PMID- 16716933 TI - Synthesis and structure analysis of novel disaccharides containing D-psicose produced by endo-1,4-beta-D-xylanase from Aspergillus sojae. AB - Xylosylpsicoses (XPs) were synthesized by transglycosylation (TG) with arabinoxylan (AX) and D-psicose (D-psi) using endo-1,4-beta-D-xylanase (EC 3.2.1.8) from Aspergillus sojae. The structures of XPs were analyzed by HPLC to determine sugar composition and molecular mass, by methylation analysis using GC MS to determine linkage, and by 1H- and 13C-NMR spectrometries to obtain the anomeric configuration of glycosidic linkage. By chemical analysis, it was found that the structures of XPs are Xylp beta1-4Psip, Xylp beta1-5Psip, Xylp beta1 1Psif and Xylp beta1-6Psif. PMID- 16716934 TI - Generation of monoclonal antibodies using simplified single-cell reverse transcription-polymerase chain reaction and cell-free protein synthesis. AB - The single-step PCR amplification of IgG Light chain (Lc) and Heavy chain (Hc) (Fd portion) from the cDNAs of a single cell was facilitated using a low concentration of cDNA-specific primers with 5' homotags in the presence of a homotag-specific primer. This method was found to be successful in generating a functional antibody with an antigen-binding activity and useful for the high throughput generation or screening of monoclonal antibodies. PMID- 16716936 TI - Community dynamics of methanotrophic bacteria during composting of organic matter. AB - In this study, we describe the effects of composting on the diversity, abundance and activity of the methanotrophic community present in the compost. Composting was allowed to proceed for 10 weeks in an in-vessel reactor. Self-heating capacity (Rottegrad) indicated that compost maturity was reached after 4 weeks. After 6 weeks, a second thermophilic phase was induced by manually increasing temperature to investigate whether or not the methanotrophs shifted back to the thermophilic population. Polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) with primers specific for type I methanotrophs of 16S rDNA and 16S rRNA were used to characterize the composition of the microbial community. Cluster and diversity analyses of RNA DGGE patterns were more sensitive than those of DNA DGGE patterns, and revealed that mesophilic and thermophilic methanotrophic communities could be differentiated. Moreover, it was seen that the diversity of the community was low during the thermophilic phase and increased during the final maturation phase. Real-time PCR analysis was also performed on the DNA and RNA extracts and showed no changes in the abundance of type I methanotrophs during the composting process (10(9) DNA copies/g compost). However, RNA-related activity did change, with the lowest activity (10(7) cDNA copies/g compost) observed during the thermophilic phase, subsequently increasing to its maximum value (10(9) cDNA copies/g compost), and finally decreasing during the maturation phase. This study confirmed the population dynamics, as seen for general groups such as bacteria and fungi during composting, for a very specific and sensitive group of bacteria, it is the type I methanotrophs. PMID- 16716935 TI - Metabolic engineering of cell cultures versus whole plant complexity in production of bioactive monoterpene indole alkaloids: recent progress related to old dilemma. AB - Monoterpene indole alkaloids (MIAs) are a large class of plant alkaloids with significant pharmacological interest. The sustained production of MIAs at high yields is an important goal in biotechnology. Intensive effort has been expended toward the isolation, cloning, characterization and transgenic modulation of genes involved in MIA biosynthesis and in the control of the expression of these biosynthesis-related genes. At the same time, considerable progress has been made in the detailed description of the subcellular-, cellular-, tissue- and organ specific expressions of portions of the biosynthetic pathways leading to the production of MIAs, revealing a complex picture of the transport of biosynthetic intermediates among membrane compartments, cells and tissues. The identification of the particular environmental and ontogenetic requirements for maximum alkaloid yield in MIA-producing plants has been useful in improving the supply of bioactive molecules. The search for new bioactive MIAs, particularly in tropical and subtropical regions, is continuously increasing the arsenal for therapeutic, industrially and agriculturally useful molecules. In this review we focus on recent progress in the production of MIAs in transgenic cell cultures and organs (with emphasis on Catharanthus roseus and Rauvolfia serpentina alkaloids), advances in the understanding of in planta spatial-temporal expression of MIA metabolic pathways, and on the identification of factors capable of modulating bioactive alkaloid accumulation in nontransgenic differentiated cultures and plants (with emphasis on new MIAs from Psychotria species). The combined use of metabolic engineering and physiological modulation in transgenic and wild-type plants, although not fully exploited to date, is likely to provide the sustainable and rational supply of bioactive MIAs needed for human well being. PMID- 16716937 TI - Identification of enzyme responsible for erythritol utilization and reaction product in yeast Lipomyces starkeyi. AB - We have identified the enzyme responsible for erythritol utilization and its reaction product in the yeast Lipomyces starkeyi CBS 1807. The enzyme, a polyol dehydrogenase requiring NAD+ as a coenzyme, was induced by erythritol in this yeast. We confirmed that the enzyme product was L-erythrulose by MS, NMR, and polarimeter analyses, meaning that we clarified the first step of erythritol utilization in yeasts for the first time. In the case of the oxidative reaction, D-threitol, (2R,3R)-2,3-butanediol, and erythritol were much better substrates than 21 other polyols tested. These three substrates are tetroses and have an R configuration at C-3, and whose third carbon results in easiest oxidation in this enzyme. The research of the substrate specificity in the reductive reaction demonstrated that L-erythrulose and dihydroxyacetone were better substrates, that D-acetoin was inactive and L-erythrose (aldose) was slightly active. PMID- 16716938 TI - Removal of hydrogen sulfide by sulfate-resistant Acidithiobacillus thiooxidans AZ11. AB - Toxic H2S gas is an important industrial pollutant that is applied to biofiltration. Here, we examined the effects of factors such as inlet concentration and space velocity on the removal efficiency of a bacterial strain capable of tolerating high sulfate concentrations and low pH conditions. We examined three strains of Acidithiobacillus thiooxidans known to have sulfur oxidizing activity, and identified strain AZ11 as having the highest tolerance for sulfate. A. thiooxidans AZ11 could grow at pH 0.2 in the presence of 74 g l( 1) sulfate, the final oxidation product of elemental sulfur, in the culture broth. Under these conditions, the specific sulfur oxidation rate was 2.9 g-S g DCW (dry cell weight)(-1) d(-1). The maximum specific sulfur oxidation rate of A. thiooxidans AZ11 was 21.2 g-S g-DCW(-1) d(-1), which was observed in the presence of 4.2 g-SO4(2-) l(-1) and pH 1.5, in the culture medium. To test the effects of various factors on biofiltration by this strain, A. thiooxidans AZ11 was inoculated into a porous ceramic biofilter. First, a maximum inlet loading of 670 g-S m(-3) h(-1) was applied with a constant space velocity (SV) of 200 h(-1) (residence time, 18 s) and the inlet concentration of H2S was experimentally increased from 200 ppmv to 2200 ppmv. Under these conditions, less than 0.1 ppmv H2S was detected at the biofilter outlet. When the inlet H2S was maintained at a constant concentration of 200 ppmv and the SV was increased from 200 h(-1) to 400 h(-1) (residence time, 9 s), an H2S removal of 99.9% was obtained. However, H2S removal efficiencies decreased to 98% and 94% when the SV was set to 500 h(-1) (residence time, 7.2 s) and 600 h(-1) (residence time, 6 s), respectively. The critical elimination capacity guaranteeing 96% removal of the inlet H2S was determined to be 160 g-S m(-3) h(-1) at a space velocity of 600 h(-1). Collectively, these findings show for the first time that a sulfur oxidizing bacterium has a high sulfate tolerance and a high sulfur oxidizing activity below pH 1. PMID- 16716939 TI - Characterization of monofunctional catalase KatA from radioresistant bacterium Deinococcus radiodurans. AB - Catalase plays a key role in protecting cells against toxic reactive oxygen species. Here we report on the cloning, purification and characterization of a catalase (KatA, DR1998) from the extremely radioresistant bacterium Deinococcus radiodurans. The size of purified D. radiodurans KatA monomer was 65 kDa while gel filtration revealed that the size of the enzyme was 240 kDa, suggesting that KatA formed a homotetramer in solution. Purified KatA displayed a final specific activity of 68,800 U/mg of protein. The catalase activity of KatA was inhibited by sodium azide, sodium cyanide and 3-amino-1,2,4-triazole. The absorption spectrum of KatA exhibited a Soret band at 408 nm. The position of the spectral peak remained unchanged following reduction of KatA with dithionite. No peroxidase activity was found for KatA. These results demonstrate that D. radiodurans KatA is a typical monofunctional heme-containing catalase. The stability of KatA with respect to H2O2 stress was superior to that of commercially available Aspergillus niger and bovine liver catalases. The relative abundance of KatA in cells in addition to the H2O2 resistance property may play a role in the survival strategy of D. radiodurans against oxidative damage. PMID- 16716940 TI - Biodesulfurization of alkylated forms of dibenzothiophene and benzothiophene by Sphingomonas subarctica T7b. AB - Sphingomonas subarctica T7b was isolated from soil in Toyotomi, Hokkaido, Japan as an organism capable of desulfurizing aromatic hydrocarbons in light gas oil (LGO) through enrichment culture. S. subarctica T7b could grow on mineral salt sulfur-free (MSSF) medium with the n-tetradecane oil phase containing dibenzothiophene (DBT), alkyl dibenzothiophenes (alkyl DBTs) or alkyl benzothiophenes (alkyl BTs) as the sole sulfur source and desulfurize these compounds, but could not utilize the tetradecane as a carbon source. This is the first report of a gram-negative bacterium which can desulfurize 4,6-dibutyl DBT and 4,6-dipentyl DBT. The desulfurized product of DBT produced by this strain was 2-hydroxybiphenyl, as in the case of other DBT-desulfurizing bacteria. S. subarctica T7b could desulfurize LGO and the sulfur content was decreased to 41% within 36 h. PMID- 16716941 TI - Lipase localization in Rhizopus oryzae cells immobilized within biomass support particles for use as whole-cell biocatalysts in biodiesel-fuel production. AB - To identify the lipase responsible for the methanolysis activity of fungus whole cell biocatalysts, the lipase localization of Rhizopus oryzae cells was determined. Western blot analysis showed that R. oryzae cells produce two types of lipase with different molecular masses of 34 and 31 kDa; the former (ROL34) was bound to the cell wall, whereas the latter (ROL31) was mainly bound to the cell membrane. It was found that cell immobilization within reticulated polyurethane foam biomass support particles strongly inhibits the secretion of membrane-bound lipase into the culture medium. An investigation of the relationship between ROL34 and ROL31 suggested that ROL31 originates from the cleavage of a 28-amino-acid residue at the N-terminus of ROL34. The addition of olive oil to the culture medium led to the retention of increased amounts of lipase within the cell. This phenomenon was further confirmed by an immunofluorescence labeling of hyphal cells. When cells were cultivated with various substrate-related compounds, such as olive oil and oleic acid, the intracellular methanolysis activity strongly correlated with the relative amounts of the membrane-bound lipase, which suggests that ROL31 localized in the membrane plays a crucial role in the methanolysis activity of R. oryzae cells. PMID- 16716942 TI - DNA taken into Bacillus subtilis competent cells by lysed-protoplast transformation is not ssDNA but dsDNA. AB - Competent Bacillus subtilis incorporates whole-genome DNA (4215 kb) from the protoplast lysate of B. subtilis subtilis [Akamatsu, T. and Taguchi, H., Biosci. Biotechnol. Biochem., 65, 823-829 (2001)]. A continuous incorporated DNA is longer than 1500 kb [J. Biosci. Bioeng., 101, 257-262 (2006)]. Whether the incorporated DNA is single-stranded (ssDNA) or double-stranded DNA (dsDNA) has been studied by examining the transforming activity of the incorporated DNA. B. subtilis BEST7027 was used as the donor strain, which has a heterologous region consisting of the 145 kb region of the Synechocystis sp. PCC6803 genome and erm gene. The donor DNA was transferred to a wild-type or a recA recipient strain (AYG2 or SYN9), and protoplast lysate was prepared from the transformants and used as the donor DNA source for the second recipient strain (AU1 or AV1). The intergenote region showed a significant transforming activity. When DNase I was added to both cells collected from the first transformation mixture and the following protoplastization, the result was similar to that obtained without DNase I. All of the observations strongly suggest that the incorporated DNA is dsDNA, and the transformation of competent B. subtilis by DNA in protoplast lysate is different from that by purified DNA taken up conventionally. PMID- 16716943 TI - Efficient biosynthesis of D-allose from D-psicose by cross-linked recombinant L rhamnose isomerase: separation of product by ethanol crystallization. AB - Mass production of a rare aldohexose D-allose from D-psicose was achieved in a batch reaction by crude recombinant L-rhamnose isomerase (L-RhI) cross-linked with glutaraldehyde. The D-psicose substrate was, in turn, mass produced from a naturally abundant ketohexose D-fructose by immobilized recombinant D-tagatose 3 epimerase (D-TE). At an equilibrium state, 25% of D-psicose was isomerized to D allose, that is, 25 g of D-allose was obtained from 100 g of D-psicose. The D allose product was easily separated and crystallized from the reaction mixture that contains 25%D-allose, 8%D-altrose and 67%D-psicose using ethanol. Empirically, approximately 338 g, that is, 90% of a theoretical overall yield for the purification of pure D-allose crystals was produced from 1.5 kg of D-psicose within 30 d using a constructed bioreactor. The cross-linked enzyme had an operative half-life of two months after repeated usages. PMID- 16716944 TI - Accelerated aromatic compounds degradation in aquatic environment by use of interaction between Spirodela polyrrhiza and bacteria in its rhizosphere. AB - Accelerated degradation of organic chemicals by aquatic plant-bacterial associations was reported for the first time with elucidation of the role and contribution of aquatic plant and bacteria in its rhizosphere using a fast growing giant duckweed, Spirodela polyrrhiza. The results clearly showed the accelerated degradation of all the three aromatic compounds (phenol, aniline and 2,4-dichlorophenol [2,4-DCP]) tested by aquatic plant-bacterial associations. In phenol degradation system, phenol-degrading bacteria indigenous to the rhizosphere fraction of S. polyrrhiza mainly contributed, while in aniline degradation system S. polyrrhiza mainly contributed by stimulating aniline degrading bacteria both in the rhizosphere and balk water fraction. On the other hand in 2,4-DCP degradation system, S. polyrrhiza itself mainly contributed to its removal by uptake and degradation. Thus, the mechanisms for accelerated removal of aromatic compounds were quite different depending on the substrates. S. polyrrhiza showed selective accumulation of phenol-degrading bacteria in its rhizosphere fraction, while aniline- and 2,4-DCP-degrading bacteria were not much accumulated. S. polyrrhiza secreted peroxidase and laccase. However, both of the enzymatic activities increased with the addition of aromatic compounds, degrading ability of S. polyrrhiza itself should be owing to the production of peroxidase rather than laccase because the change of peroxidase activity and concentration of each aromatic compound well concurred. From the results obtained in the present study, it can be concluded that the feasibility of the use of aquatic plant-bacterial associations to accelerate the degradation of organic chemicals especially recalcitrant compounds in aquatic environment was shown. PMID- 16716945 TI - Construction of an additional metal-binding site in human metallothionein-2. AB - We have constructed a new metal-binding site in the human metallothionein-2 (hMT 2), using the protein as a scaffold to investigate the structure and function of metal-binding. Potential metal-binding sites were designed within hMT-2 on the basis of structures generated by homology modeling. Amino acid residues D11, C13, C26 and S28 in the beta-domain of hMT-2 (hMT-2beta) were found, by computer search, to form a potential tetrahedral Cys4 metal-binding site. Six mutant proteins were constructed with the following amino acid substitutions: D11C, S28C and D11C/S28C in hMT-2 and the same mutations in hMT-2beta, respectively. These single-mutant and double-mutant proteins bound one gram atom of cadmium or zinc ions per gram molecule of protein more than the corresponding wild-type proteins. The circular dichroism spectra suggested that the structures of the single-mutant proteins that bound Cd or Zn were similar to that of the D11C/S28C double-mutant proteins. To evaluate the metal-binding affinity of the mutant proteins, we performed pH titrations of wild-type and mutant proteins. The stability with changes in pH of all the mutant proteins was higher than that of the wild-type proteins, and that of the double-mutant D11C/S28C protein was highest. Consequently, it appears that we were able to create novel proteins that bound metal ions at high density and with high affinity. PMID- 16716946 TI - Characterization of transient expression system for retroviral vector production. AB - The production of retroviral vectors using a transient expression system has been improved to obtain a high-titer virus preparation that is difficult to produce using packaging cell lines due to the cytotoxic or cytostatic effect of transgenes. Here, we used one such production method, the so-called Q-vector system, and examined its potential for virus production. The Q-vector system could produce a similar level of viral vectors compared with the packaging cell system but the production seemed to depend on the size and nature of transgenes. In the process of investigation of the quantitative difference in viral components between the transient expression system and the packaging cell system, we found that the Q-vector system could express higher amounts of viral RNA and proteins compared with the packaging cell system. However, this did not lead to a higher virus titer compared with that produced by the packaging cell system. This suggests that retroviral RNA transcribed from the plasmid in the transient system seemed to be used mainly for translation and only some of the RNA molecules were packaged in viral particles. PMID- 16716947 TI - Improved microcirculatory effect of D-allose on hepatic ischemia reperfusion following partial hepatectomy in cirrhotic rat liver. AB - D-allose, one of the rare sugars produced from D-psicose, has been shown to be effective against reperfusion injury after ischemia and partial hepatectomy in cirrhotic rat liver by improving remnant liver blood flow and survival rates, and decreasing liver enzyme levels and liver tissue injury levels. These findings demand further study of the clinical implications of this sugar in view to the advancing fields of liver surgery and transplantation. PMID- 16716948 TI - Functional expression of streptococcal galactosyltransferase in baculovirus/insect cell expression system. AB - The cpsIaJ gene of Streptococcus agalactiae type Ia codes for beta-1,4 galactosyltransferase. In this study, the functional expression of His-tagged CpsIaJ in a baculovirus expression system was performed, because the efficient functional expression of this enzyme in Escherichia coli had been unsuccessful. Using a partially purified enzyme preparation, we found that the enzyme had a restricted substrate specificity and that the entire structure of the substrate GlcNAc beta1-3Gal beta1-4Glc was required for the activity. Furthermore, mutations in a conserved DXD motif caused the loss of the enzyme's activity. PMID- 16716949 TI - Perforator flaps. PMID- 16716950 TI - Breast reconstruction with the deep inferior epigastric perforator flap: history and an update on current technique. AB - Perforator flaps allow the transfer of the patient's own skin and fat in a reliable manner with minimal donor site morbidity. For breast reconstruction, the abdomen typically is our primary choice as a donor site. The deep inferior epigastric perforator (DIEP) flap remains our first choice as an abdominal perforator flap and has become a mainstay for the repair of mastectomy defects. It allows the transfer of the same tissue from the abdomen to the chest for breast reconstruction as the TRAM flap without sacrifice of the rectus muscle or fascia. We discuss our current techniques and specific issues related to the surgery. We present the results of 1095 cases of free tissue transfers from the abdomen for reconstruction of the breast. PMID- 16716951 TI - Preoperative planning of deep inferior epigastric artery perforator flap reconstruction with multislice-CT angiography: imaging findings and initial experience. AB - BACKGROUND: Autologous breast reconstruction with abdominal tissue is one of the best options after mastectomy. Reconstruction with deep inferior epigastric perforator (DIEAP) flaps requires a precise location and preoperative evaluation of perforating vessels. The objective of this report is to demonstrate the usefulness of multislice-CT (MSCT) angiography for preoperative planning in patients undergoing DIEAP flap reconstruction. METHODS: Six consecutive women were considered for breast reconstruction with DIEAP flaps after previous mastectomy for breast cancer. Preoperative MSCT angiography was performed to localise the arterial perforators. Axial images, multiplanar reconstructions (MPR) and 3D volume-rendered images were analysed. Findings were correlated with surgery. Initial experience and imaging findings will be described. RESULTS: Accurate identification of the main perforators was achieved in all six patients with a very satisfactory concordance between MSCT angiography and surgical findings. No unreported vessels were found. Location, course, anatomical variations and relations of the superficial inferior epigastric artery were reported. The very small perforators, were equally evaluated and described. CONCLUSIONS: Preoperative evaluation of perforator arteries with MSCT angiography is feasible in patients undergoing breast reconstruction. This technique provides a noninvasive global approach of the vascular anatomy and the entire anterior abdominal wall. However, more patients need to be evaluated in order to clarify the potential aspects pointed in this report. PMID- 16716952 TI - Multidetector-row computed tomography in the planning of abdominal perforator flaps. AB - An accurate preoperative evaluation of the vascular anatomy of the abdominal wall is extremely valuable in improving the surgical strategy in abdominal perforator flaps. The multidetector-row computer tomography offers thin slice coverage of extended volumes with an extremely high spatial resolution. From October 2003 to December 2004, 66 female patients had breast reconstruction surgery in our department using the deep inferior epigastric artery perforator flap. Our multidetector-row computer tomography studies were performed using a 16-detector row computer tomography scanner. The image assessment was carried out using the following protocol: we first identified the best three perforators from each side of the abdomen. Then we conducted a three-dimensional reconstruction of the abdomen by identifying exactly where the three best perforators emerged from the rectus abdominis fascia. We then transferred the data obtained from the image to the patient using a coordinate system. In addition, we also placed the dominant perforators in the patient by using a conventional hand-held Doppler. During the operation we compared intra-operative findings, Doppler results and computer tomography outcomes. Neither false positive nor false negative results were found in the computer tomography outcome. Multidetector-row computer tomography provides us with an easy method of interpreting the virtual anatomic dissection in three dimensions. It has high sensitivity and specificity and provides a good quality evaluation of the perforator vessels. This information allows reduction of operating time and safer performance of surgery. The multidetector-row computer tomography is a highly effective tool in the preoperative study of abdominal perforator flaps. PMID- 16716953 TI - Microcirculation in DIEP flaps: a study of the haemodynamics using laser Doppler flowmetry and lightguide reflectance spectrophotometry. AB - The deep inferior epigastric perforator (DIEP) flap is ideally suited for breast reconstruction. The DIEP flap reduces abdominal wall morbidity when compared to a transverse rectus abdominis (TRAM) flap, however, there maybe a compromise of the flap vascularity. Early venous problems and late fat necrosis in DIEP flaps can occur. There is a limited literature on the haemodynamic and microcirculation of the DIEP flap. Sixteen consecutive patients who underwent immediate breast reconstruction with DIEP flap were prospectively monitored during the post operative time for a minimum period of 48h using laser Doppler flowmetry (LDF) and lightguide reflectance spectrophotometry (LRS) to analyse blood flow and oxygenated haemoglobin percentage in the cutaneous microcirculation of the flap. LDF demonstrated an increase of capillary flow in comparison to the pre-operative levels. LRS showed an initial decrease of the values and a varying pattern of increase in relation to time, to reach the pre-operative levels. A correlation between lower values of post-operative LDF and LRS measurements and late fat necrosis and early venous congestion was demonstrable in this series. This study demonstrates a change in the cutaneous microcirculation of the lower anterior abdominal wall following its elevation as a DIEP flap. These findings could explain the clinical phenomenon occurring in this flap in the early post operative period. Significant correlation between longer harvesting time and re exploration and between longer ischaemia time and localised fat necrosis was found. PMID- 16716955 TI - Breast reconstruction with gluteal artery perforator flaps. AB - BACKGROUND: Several alternatives exist for breast cancer reconstruction with perforator flaps. For those patients in whom the buttock is the best choice as a source for autologous tissue, the IGAP and SGAP flaps are an excellent option. These flaps allow the reliable transfer of skin and soft tissue from the buttock without the associated donor site morbidity of a muscle flap. INDICATIONS: Most women requiring tissue transfer to the chest from the buttock for breast reconstruction or other reasons are candidates for IGAP or SGAP flaps. Do to an improved donor site contour and scar, we now prefer to use the IGAP to the SGAP flap. Absolute contraindications specific to perforator flap breast reconstruction in our practice include history of previous liposuction of the donor site or active smoking (within 1 month prior to surgery). ANATOMY AND TECHNIQUE: IGAP and SGAP flaps are based on perforators from either the superior or inferior gluteal artery. These perforators are carefully dissected free from the surrounding gluteus maximus muscle, which is spread in the direction of the muscle fibres and safely preserved. The vascular pedicle is anastomosed to recipient vessels in the chest and the donor site closed primarily. CONCLUSIONS: IGAP and SGAP flaps allow the safe and reliable transfer of tissue from the buttock for breast reconstruction as an alternative to soft tissue transfer from an abdominal donor site or even as a first choice in selected patients. PMID- 16716956 TI - Reconstruction of skull base and dura using rectus abdominis muscle combined with a vascularised fascial perforator flap. AB - The goal of reconstruction of the skull base is to repair dural defects with a watertight seal and separate it from the nasopharynx and the exterior environment with well vascularised tissue. This prevents cerebro-spinal fluid leakage and secondary contamination leading to the potentially life threatening complications of meningitis or extradural abscess. Following large composite resections, traditional techniques to repair the dura involve the use of an autologous fascial graft or a pericranial flap, whilst a regional pedicled or free muscle flap is used to close the dead space defect. We describe a technique performed in two cases, whereby a single flap, the rectus abdominis muscle free flap, can be used to provide vascularised reconstruction both of the dura and the skull base. The anterior rectus sheath, islanded on a single perforator vessel, is used as a vascularised layer to reconstruct dura, whilst the supporting rectus abdominis muscle provides bulk to obliterate dead space. We show that this flap is suitable for reconstruction even in the presence of chronic infection. Advantages of a vascularised reconstruction are the rapid healing of the wound, even after radiotherapy, the delivery of systemic antibiotics to the site of the operation, and that it may allow early postoperative radiotherapy to be planned. PMID- 16716957 TI - The versatility of the inter-costal artery perforator (ICAP) flaps. AB - INTRODUCTION: Anatomy and classification of intercostal perforator flaps in addition to our experience with will be demonstrated for different indications. MATERIAL AND METHODS: The intercostal vessels form an arcade between the aorta and the internal mammary vessels. Different pedicled perforator flaps can be raised on this neurovascular bundle to cover defects on the trunk. They are classified as following: dorsal intercostal artery perforator flap (DICAP); lateral intercostal artery perforator (LICAP); and anterior intercostal artery perforator (AICAP) flap. RESULTS: Between 2001 and 2004, 20 pedicled (ICAP) flaps were harvested in 16 patients. The indications were: immediate partial breast reconstruction in eight patients who had a quadrantectomy for breast cancer; midline back and sternal defects in three patients who had radical excisions for a dermatofibrosarcoma or malignant melanoma; and autologous breast augmentation (four bilateral and one unilateral flap) in five post-bariatric-surgery patients. The average flap dimension was 18 x 8 cm(2) (range 8 x 5-24 x 12 cm(2)). There were two DICAP flaps, two (AICAP) flaps and 16 (LICAP) flaps. All but two flaps were based on one perforator. Mean harvesting time was 45min for a single flap. Bilateral breast augmentation with LICAP flap necessitated longer operative time (range 2-3h) depending whether it was combined or not with mastopoexy. Complete flaps survival was obtained. All donor sites were closed primarily. CONCLUSION: The (ICAP) flaps provide valuable options in breast surgery; and for challenging defects on the trunk without sacrifice of the underlying muscle. PMID- 16716958 TI - Flap-in-flap method for enhancing the advancement of a V-Y flap. AB - The V-Y principal is used in a number of flap reconstruction techniques. V-Y flaps suffer from design problems of the advancing edge, which is usually concave, trying to match another concave edge on the opposite side of the defect. The flap-in-flap technique uses a second V-Y flap at the advancing edge of the main flap to overcome this mismatch, and also improves the amount of advancement possible. The technique has been used in three patients, demonstrating reliability and a further advantage, which is the ability of the second flap to rotate at an angle to the main flap to aid in reconstructing a complex defect such as the alar base of the nose. Further experience with this technique should widen its applications. PMID- 16716959 TI - Lower limb salvage to allow planned trans-tibial amputation. AB - Early, primary amputations are still necessary for certain patients sustaining high impact lower limb trauma. For cases with extensive proximal tibial bone loss a turn-up bone flap technique has been described to achieve a below-knee stump suitable for a prosthesis. However, in certain circumstances, for example if posterior soft tissues are injured, this type of reconstruction is not possible. This case report demonstrates that converting a severe open proximal tibial fracture to a successful below-knee amputation is also possible with acute limb shortening, flap cover and planned subsequent trans-tibial amputation. PMID- 16716960 TI - Stenting the pancreatic duct. PMID- 16716961 TI - Rectal cancer: from outcomes of care to process of care. AB - This paper represents a current opinion on the impact surgeons may have on the variability of the quality of care of rectal cancer surgery. No systematic review of the evidence available in the literature is provided. The objective is to present a concise insight on selected outcomes of care studies, to review the limitations of such studies and to discuss the value of process of care studies. Outcomes of care studies measure what happens to patients, and process of care studies measure what is done to patients. Three variables are reviewed: training, volume and individual skill. It is concluded that the quality of the selected outcomes of care studies is not sufficient to draw definitive conclusions on whether surgeons are a variable. Further efforts should prompt process of care studies on rectal cancer surgery. This implies that outcomes should be measured, processes of care modified and outcomes measured again. This cycle should be continuously repeated in order to achieve the best quality of care. PMID- 16716962 TI - Prevalence, comorbidity and impact of irritable bowel syndrome in Norway. AB - OBJECTIVE: To study the prevalence of irritable bowel syndrome (IBS) and its comorbidity in a Norwegian adult population. MATERIAL AND METHODS: In 2001, 11,078 inhabitants (aged 30-75 years) in Oppland County were invited to take part in a public health survey. A total of 4622 subjects (42%) completed the questionnaires on symptoms of IBS (Rome II criteria), comorbidity, health-care visits and medications. The impact of comorbidity on global health, working disability and use of health-care resources in subjects with IBS was explored by stepwise logistic regression. RESULTS: The population prevalence of IBS was 388/4622 (8.4% (95% CI: 7.6-9.4%)) with a female predominance and an age dependent decrease. The proportion who had consulted for IBS ranged from 51% among 30-year-olds to 79% in 75-year-olds (p=0.05). IBS was associated with musculoskeletal complaints (OR = 2.4-3.4 for six different items), fibromyalgia (OR = 3.6 [2.7-4.8]), mood disorder (OR = 3.3 (2.6-4.3)), reduced global health (OR = 2.6 (2.1-3.2)), working disability (OR = 1.6 (1.2-2.1)), more frequent health-care visits and use of medications (OR 1.7-2.3). When controlling for comorbidity, reduced global health (OR = 1.5 (1.1-2.0)) and use of alternative health care (OR = 1.7 (1.3-2.4)) remained associated with IBS. Severity of abdominal pain/discomfort was a predictor of having to seek a physician for IBS (OR = 1.3 (1.2-1.5)). CONCLUSIONS: Symptoms of IBS were reported by 8% of Norwegian adults and had resulted in consultations with physicians for the majority in the long run. Subjects with IBS in the community were characterized by frequent somatic and psychiatric comorbidity. Their observed reduced health, working disability and increased use of health resources were largely explained by comorbid symptoms and disorders. PMID- 16716963 TI - Low social class is linked to upper gastrointestinal symptoms in an Australian sample of urban adults. AB - OBJECTIVE: The epidemiology of gastrointestinal (GI) symptoms has been described in population surveys, yet their distribution by socio-economic (social) class remains largely uninvestigated. The aim of this study was to evaluate the influence of social class on GI symptoms in an urban sample of Australian adults. MATERIAL AND METHODS: The prevalence of 25 GI symptoms was determined by postal questionnaire. Five latent symptom groups were identified by a principal components analysis (PCA) (Esophageal, Dysmotility-like, Nausea/vomiting, Constipation and Diarrhea). These components were used to model the association between GI symptoms and adult social class. Social class was assigned according to a census-based measure of area disadvantage, and to highest level of completed education. Age- and sex-adjusted odds ratios - as identified by unconditional logistic regression - were used to describe the relationship between symptom groups and adult social class. RESULTS: The effects of area disadvantage and education on Esophageal and Dysmotility-like symptoms were pronounced, with persistent trends for elevated symptom rates amongst the lower social classes (all p<0.01 on age- and sex-adjusted effects). When defined by area disadvantage, the odds ratios for Nausea/vomiting were significantly elevated among the lowest social class group (p=0.01), whereas the odds for Constipation were significantly elevated among the upper-middle social class when defined by education (p=0.001). Diarrhea was not associated with social class whether defined by area disadvantage or education. CONCLUSIONS: Low social class is a risk factor for upper GI complaints. PMID- 16716964 TI - Inhibitory effect of GLP-1 on gastric motility persists after vagal deafferentation in pigs. AB - BACKGROUND: Glucagon-like peptide 1 (GLP-1) is an intestinal hormone that is secreted in response to meal ingestion. GLP-1 inhibits gastric emptying and reduces postprandial gastric secretion and may play a physiological regulatory role in controlling appetite and energy intake in humans. The GLP-1 receptors have been identified in several organs including the stomach, brain and pancreas. The GLP-1 mechanism of action on insulin secretion is at least partly mediated via receptors on the pancreatic islet, but the mechanism by which GLP-1 retards gastric emptying is not known and may involve neural interactions, although GLP-1 has no effect on vagally stimulated motor activity of the isolated porcine antrum. MATERIAL AND METHODS: Previously, an experimental model was developed with centrally (insulin hypoglycaemia) induced vagally mediated stimulation of antral motility, recorded by force transducers, in anaesthetized pigs. This model has now been developed further to include vagal deafferentation to determine the role of the afferent vagus in mediating the inhibitory effect of GLP-1 on gastric motility. RESULTS: Intravenous infusion of GLP-1 resulting in slightly supraphysiological plasma levels inhibited the antral contractile force, with the amplitude falling from 29.9+/-5.7 mm to 14.6+/-3.5 mm (p<0.001). After vagal deafferentation GLP-1 still inhibited antral motility (from 36.6+/-6.4 mm to 25+/ 4.4 mm (p<0.019). The decrease in amplitude was the same before and after deafferentation. CONCLUSIONS: GLP-1 significantly inhibited centrally induced antral motility and the inhibitory effect of GLP-1 on gastric motility persisted after vagal deafferentation, supporting the hypothesis that the inhibitory effect results from direct interaction of GLP with receptors in the CNS, which in turn reduce vagal efferent output. PMID- 16716965 TI - Vitamin D deficiency in patients operated on for gastric lymphoma. AB - OBJECTIVE: To determine the nutritional status in patients treated for gastric non-Hodgkin's lymphoma (NHL). MATERIAL AND METHODS: Patients treated during the period 1990-99 according to a protocol including primary gastric surgery for eligible patients were registered prospectively in successive patients. Those with aggressive lymphomas in stage IE-IIE underwent gastric surgery followed by CHOP-like chemotherapy. Patients with indolent lymphomas and localized disease did not receive any further treatment if the operation was considered radical; otherwise, they received local radiotherapy after surgery. Patients with advanced disease underwent gastric surgery only if there was a considerable risk of bleeding or perforation. Patients below the age of 80 years and in complete remission were offered a cross-sectional examination a median of 102 months later, including clinical examination with estimation of body mass index, upper endoscopy and blood tests (haemoglobin, ferritin, serum-Fe, total iron-binding capacity (TIBC), vitamin B(12), homocysteine, vitamin D status, parathyroid hormone (PTH), albumin and electrolytes). RESULTS: Forty patients were identified, of whom 33 met for follow-up examination. Seventeen patients had a partial gastrectomy (PG), 9 a total gastrectomy (TG) and 7 patients were not operated on. The patients in the TG group had significant weight loss. Furthermore, the patients in the TG group had a lower storage iron content (s ferritin and s-iron saturation), lower s-vitamin D, higher s-PTH and homocysteine than the other groups. CONCLUSIONS: If surgery is necessary for gastric lymphomas, a PG should be performed when possible. The patients should receive dietary advice and be followed-up at least yearly for nutritional deficiencies. Regular intake of vitamin D and -B(12), calcium, folate and iron should be considered. PMID- 16716966 TI - ColoPlus, a new product based on bovine colostrum, alleviates HIV-associated diarrhoea. AB - OBJECTIVE: HIV-associated diarrhoea occurs in nearly all patients with acquired immunodeficiency syndrome (AIDS) in the developing countries. Diarrhoea is caused by the HIV-related immune dysfunction and is pivotal in the decrease of the helper T-cell (CD4 + ) population. Enteric pathogens in HIV-associated diarrhoea are, for example, Cryptosporidium, Amoeba and Campylobacter species. Bovine colostrum is the first milk the suckling calf receives from the cow. It is rich in immunoglobulins, growth factors, antibacterial peptides and nutrients. It supplies the calf with a passive immunity before its own active immunity is established. ColoPlus is a product based on bovine colostrum and is designed for slow passage through the gastrointestinal tract, as well as having a high nutritional value. The aim of the study was to investigate whether ColoPlus given orally can influence the severe diarrhoea associated with HIV infection. MATERIAL AND METHODS: The study was carried out at Braithwaite Memorial Specialist Hospital, Port Harcourt, Nigeria. Thirty patients with HIV-associated diarrhoea were included in the study. The patients were treated with ColoPlus for 4 weeks in an open-labelled non-randomized study, after an observational period of one week. After a post-treatment period of another two weeks, treatment with anti-HIV drugs was started, if deemed appropriate. The effects on the frequency of stool evacuations per day, on body-weight, fatigue, haemoglobin levels and CD4+ counts before (week 1) and after treatment with ColoPlus (week 7) were measured. RESULTS: There was a dramatic decrease in stool evacuations per day from 7.0+/ 2.7 to 1.3+/-0.5 (+/-SD), a substantial decrease in self-estimated fatigue of 81%, an increase in body-weight of 7.3 kg per patient and an increase in CD4+ count by 125%. CONCLUSION: ColoPlus may be an important alternative or additional treatment in HIV-associated diarrhoea. PMID- 16716967 TI - Vascular endothelial growth factor (VEGF164) ameliorates intestinal epithelial injury in vitro in IEC-18 and Caco-2 monolayers via induction of TGF-beta release from epithelial cells. AB - OBJECTIVE: VEGF is a glycoprotein with various (e.g. angiogenic) activities. So far, research has focused on its angiogenic properties. VEGF receptors are localized on epithelial cells of patients with inflammatory bowel disease (IBD) and also on Caco-2 and IEC-18 cells. Our aim was to evaluate the role of VEGF on intestinal epithelial cell (IEC) migration and proliferation by utilizing an established in vitro model. METHODS: IEC-18 and Caco-2 monolayers were wounded with a razor blade as described previously. Cells were incubated in medium w/o rat VEGF(164). After 24 h, migration was assessed by counting cells across the wound edge. Migration was blocked with neutralizing TGF-beta(1) antibodies. IEC proliferation was assessed using the MTT (3-[4, 5-Dimethylthiazol-2-yl]-2, 5 diphenyl-tetrazolium bromide) test. Semi-quantitative changes of the TGF-beta(1) mRNA expression were evaluated before and after stimulation of the cells with VEGF(164) by RT-PCR. Statistical analysis was performed with ANOVA and the Wilcoxon test. RESULTS: VEGF(164) significantly induced epithelial cell migration in Caco-2 and IEC-18 cells compared to control. TGF-beta(1) antibodies completely abolished this VEGF-induced cell migration. TGF-beta(1) mRNA significantly increased in IEC-18 and Caco-2 cells after stimulation with VEGF. VEGF significantly inhibited epithelial cell proliferation in IEC-18 and in Caco-2 cells, indicating that the observed effects on cell migration were not due to any proliferate effects. CONCLUSION: VEGF effects on epithelial cell migration play an important part in epithelial cell restitution by maintaining mucosal homeostasis after mucosal injury. This effect is mediated by TGF-beta(1). Our results obtain another possible role for increased VEGF levels in the intestinal mucosa of patients with IBD as reported recently by others. PMID- 16716968 TI - Norovirus outbreak in a pediatric oncology unit. AB - OBJECTIVE: Norovirus (NV) is an etiologic agent of outstanding importance that can cause severe epidemic gastroenteritis in day-care centers, schools, nursing homes, and hospitals. Therefore NV requires foremost attention as a pathogen responsible for epidemics of gastroenteritis in immunocompromised inpatients. In this study, a NV outbreak in a pediatric oncology unit is described and the consequences for this high-risk population are discussed. MATERIAL AND METHODS: Stool and vomitus samples from 11 patients were tested for NV and other relevant viruses during the outbreak by reverse transcriptase-polymerase chain reaction (RT-PCR) and/or enzyme-linked immunosorbent assay (ELISA) (whenever an appropriate ELISA was available). Norwalk virus PCR amplifications were sequenced and phylogenetic analysis was performed. RESULTS: The index patient and the chain of infection were identified. Follow-up investigation surprisingly demonstrated viral shedding for a maximum of 140 days (median 23 days). Three patients experienced severe or life-threatening symptoms, probably related to NV infection. CONCLUSIONS: In the event of an outbreak of gastroenteritis (involving two or more symptomatic patients) in a pediatric oncology unit, the search for NV in stool or vomitus specimens should be initiated in good time. As long as the data are limited regarding whether a detectable viral antigen or RNA in stools represents an infectious virus, patients have to be isolated as long as the diagnostic assays remain positive. During the acute phase of the illness, health care workers should wear masks in addition to practicing meticulous hand hygiene with a disinfectant of proven activity against NV. Pediatric oncology patients must be closely monitored during follow-up investigations as they may shed the virus for months. There is some evidence from the outbreak described here that those patients face a greater risk of severe NV-related complications. PMID- 16716969 TI - Contribution of CARD15 variants in determining susceptibility to Crohn's disease in Sweden. AB - OBJECTIVE: Crohn's disease (CD) is a chronic inflammatory bowel disorder caused by environmental and genetic factors. Mutations in the CARD15 gene have been associated with CD. No previous case-control CARD15 study has been performed in the Swedish population. MATERIAL AND METHODS: The study comprised of 321 individuals: 178 with CD and 143 healthy controls (HCs), all from Stockholm County. All were genotyped for the three main CD-associated CARD15 variants (R702W, G908R and 1007fs) and phenotypic associations were investigated. RESULTS: The allele frequencies of the R702W variant (4.5% CD versus 0.7% HC, p=0.008, OR = 6.8) and the G908R variant (2.0% CD versus 0% HC, p=0.045) were more common in CD patients than in controls. No significant difference in1007fs variant allele frequency was found between CD patients and controls (2.0% CD versus 1.7% HC, p = 0.8, OR = 1.1). Carriage of CARD15 variants was more common in the CD patients than in controls (15.2% CD versus 4.2% HC, p = 0.001, OR = 4.1, population attributable risk (PAR) = 11.4%). Genotype-phenotype analysis demonstrated that CARD15 variants were associated with ileal disease (p=0.0006, OR = 9.3, CI = 2.2 34) and protective for colonic CD (p = 0.01, OR = 0.18). An association between CARD15 variants and ileal CD (p=0.004, OR = 6.6) was confirmed by multivariate analyses. CONCLUSIONS: The CARD15 variants R702W and G908R, but not 1007fs, are associated with susceptibility to CD in Stockholm County. Genotype-phenotype analysis shows an association with ileal CD. The contribution of these CARD15 mutations in Swedish CD patients overall is low in relation to studies elsewhere in Central Europe and North America, but is consistent with emerging data from elsewhere in Scandinavia and in Northern Europe. PMID- 16716970 TI - Cytomegalovirus infection in ulcerative colitis. AB - OBJECTIVE: Cytomegalovirus (CMV) infection has been reported as an exacerbating factor in inflammatory bowel disease but the relationship between CMV infection and ulcerative colitis (UC) remains unclear. There has been no detailed research to elucidate the clinicopathologic features of CMV infection in UC using surgical specimens. The aim of this study was to investigate the clinicopathologic features of CMV infection in UC patients who had undergone colectomy. MATERIAL AND METHODS: Surgical specimens taken from UC patients were examined for CMV infection. The patients were divided into three groups: severe, refractory, and UC-associated dysplasia or cancer according to the operative indications. CMV infection rates were evaluated and a comparison of clinical parameters was made between CMV-positive and CMV-negative patients, and the risk factors for CMV infection were analyzed using multivariate analyses. RESULTS: It was found that 25% of 32 patients were positive for CMV in the severe UC group; 8.3% of 72 patients were positive for CMV in the refractory UC group. None of the 22 patients was positive for CMV in the UC-associated dysplasia or cancer group. The CMV-positive rate in the severe UC group was significantly higher than that in the other groups (p<0.05). Patients' age at the time of operation was higher in the CMV-positive group than in the CMV-negative group among the patients with severe UC (p<0.01), and age at operation was an independent risk factor for CMV infection. CONCLUSIONS: CMV is found more frequently in severe UC than refractory UC and UC-associated cancer or dysplasia. Higher age can be a risk factor for CMV infection in patients with severe UC. However, a high steroid dose may not always be a risk factor for CMV infection. PMID- 16716971 TI - Histamine inactivation in the colon of pigs in relationship to abundance of catabolic enzymes. AB - OBJECTIVE: Catabolism of histamine plays a crucial role in the intestine in preventing intoxication by luminal histamine. Two enzymes are involved, namely histamine N-methyltransferase (HMT) and diamine oxidase (DAO). The purpose of this study was to find a link between histamine catabolism and the activities of HMT and DAO. MATERIAL AND METHODS: Epithelia of porcine proximal colon were mounted in Ussing chambers. After mucosal addition of (3)H-histamine (100 micromol x l(-1)) and (14)C-mannitol, the appearance of non-catabolized histamine, (3)H-histamine label (hist-rad) and (14)C-mannitol label were measured in parallel on the serosal side. Activities of HMT and DAO were determined in the proximal colon and proximal jejunum. RESULTS: Differences between the fluxes of histamine and hist-rad indicated catabolic conversion of 81.4+/-1.6% histamine during epithelial transit. Fluxes of hist-rad and histamine increased linearly with increasing mannitol fluxes but the percentage of catabolized histamine was not related to either mannitol or hist-rad fluxes. However, the percentage of catabolized histamine rose with increasing DAO activity. Given a negative correlation between DAO and HMT activities, the fraction of catabolized histamine decreased with increasing HMT activity. HMT activity was comparable in the colon and jejunum, but DAO activity was approximately nine times higher in the jejunum. CONCLUSIONS: Permeation, but not the relative efficiency of catabolism, of histamine depends on epithelial/paracellular tightness. While previous studies have shown that colonic HMT essentially catabolizes the bulk of histamine during permeation, DAO activity seems to be more variable and limiting for the overall efficiency of the catabolic process. PMID- 16716972 TI - Fecal calprotectin remains high during glucocorticoid therapy in children with inflammatory bowel disease. AB - OBJECTIVE: Fecal calprotectin is a promising marker for the assessment of gastrointestinal inflammation. Fecal calprotectin levels were followed-up in children with inflammatory bowel disease (IBD) who were introduced to glucocorticoid therapy. The aim of this study was to assess whether the changes in fecal calprotectin levels reflect therapeutic responses. MATERIAL AND METHODS: Fecal calprotectin was measured by enzyme immunoassay in 57 children (mean age 9.8 years, range 0.9-18 years) who underwent colonoscopies (IBD n=31, non-IBD disease n=13, normal n=13) and followed-up in 15 children (mean age 13 years, range 3.6-18 years) who were introduced to glucocorticoid therapy because of active IBD at 0, 2, and 4 weeks and at 4-week intervals until one month after discontinuation of the therapy. RESULTS: Fecal calprotectin was <100 microg/g in 70% of the children with normal findings on colonoscopy or a non-IBD disease. Fecal calprotectin was >100 microg/g in all but one child with active IBD and in 13/15 of those children who were introduced to glucocorticoids by the clinicians. Fecal calprotectin values decreased within 4 weeks in line with clinical improvement in 7 children and normalized in 4/15 children during the follow-up. Fecal calprotectin increased in 5/8 of the non-steroid-dependent children after discontinuation of glucocorticoids. CONCLUSIONS: Fecal calprotectin is a sensitive marker for chronic colitis. In active disease treated with glucocorticoids, fecal calprotectin levels declined in line with the clinical improvement but seldom fell within the normal range, which suggests ongoing inflammation in a clinically silent disease. The measurement of fecal calprotectin may provide new tools for the assessment of the level of gut inflammation in children with chronic colitis in the follow-up of clinical responses. PMID- 16716973 TI - Colonic mucosal tears in collagenous colitis. AB - In general, the colonic mucosa is macroscopically normal in collagenous colitis, although minor, non-specific abnormalities may be found. Significant endoscopic abnormalities, "mucosal tears" representing longitudinal mucosal lacerations, have been reported in a few patients with collagenous colitis. We report the cases of three women with collagenous colitis and mucosal tears detected at the index colonoscopy in order to illustrate the endoscopic characteristics and review the literature. Including the present cases, a total of 12 patients with mucosal tears and collagenous colitis have been reported. In 10 patients, the mucosal lacerations involved the ascending or the transverse colon. Three of the 12 patients had a colonic perforation immediately after the colonoscopy. The colonoscopist should be aware that the risk of perforation is likely to be increased when mucosal tears are present. PMID- 16716974 TI - Polyethylene glycol, unique among laxatives, suppresses aberrant crypt foci, by elimination of cells. AB - OBJECTIVE: Polyethylene glycol (PEG), an osmotic laxative, is a potent inhibitor of colon cancer in rats. In a search for the underling mechanisms, the hypothesis that fecal bulking and moisture decrease colon carcinogenesis was tested. We also investigated the PEG effects on crypt cells in vivo. MATERIAL AND METHODS: Fischer 344 rats (n=272) were injected with the colon carcinogen, azoxymethane. They were then randomized to a standard AIN76 diet containing one of 19 laxative agents (5% w/w in most cases): PEG 8000 and other PEG-like compounds, carboxymethylcellulose, polyvinylpyrrolidone, sodium polyacrylate, calcium polycarbophil, karaya gum, psyllium, mannitol, sorbitol, lactulose, propylene glycol, magnesium hydroxide, sodium phosphate, bisacodyl, docusate, and paraffin oil. Aberrant crypt foci (ACF) and fecal values were measured blindly after a 30 day treatment regimen. Proliferation, apoptosis, and the removal of cells from crypts were studied in control and PEG-fed rats using various methods, including TUNEL and fluorescein dextran labeling. RESULTS: PEG 8000 reduced the number of ACF 9-fold in rats (p<0.001). The other PEGs and magnesium hydroxide modestly suppressed ACF, but not the other laxatives. ACF number did not correlate with fecal weight or moisture. PEG doubled the apoptotic bodies per crypt (p<0.05), increased proliferation by 25-50% (p<0.05) and strikingly increased (>40-fold) a fecal marker of epitheliolysis in the gut (p<0.001). PEG normalized the percentage of fluorescein dextran labeled cells on the top of ACF (p<0.001). CONCLUSIONS: Among laxatives, only PEG afforded potent chemoprevention. PEG protection was not due to increased fecal bulking, but in all likelihood to the elimination of cells from precancerous lesions. PMID- 16716975 TI - Three randomized long-term surveillance trials in patients with sporadic colorectal adenomas. AB - OBJECTIVE: Guidelines for surveillance of patients with previous sporadic colorectal adenomas are based on retrospective long-term follow-up and prospective short-term studies. The aim of the present studies was to compare relative risk (RR) of new neoplasia as well as complications, using different intervals between examinations in long-term surveillance. MATERIAL AND METHODS: Between l98l and l991, patients with pedunculated and small, flat and sessile adenomas were allocated at random to a 24 months (group A) or 48 months (group B) interval between surveillance colonoscopies (n=671). Patients with flat and sessile adenomas greater than 5 mm in diameter were randomized to intervals of 6 months (group C) or 12 months (group D) between l981 and 1987 (n=73). Finally, 200 patients with similar adenomas as in groups C and D were randomized to 12 months (group E) or 24 months (group F) from 1988 to 2000. The study ended in 2002. RESULTS: Advanced adenomas were equally as frequent in group A and group B, but colorectal cancer (CRC) was found significantly more often in group B (RR = 6.2 (1.0-117.4)). Severe complications occurred in 4 patients in group A and 2 patients in group B. Advanced new adenomas tended to be more frequent in group D than in C (p=0.08), but only one CRC was detected and this was in group C. There was no significant difference in the risk of CRC between the E and F groups, but the two cancers in group E were both early stage, in contrast to those in group F. Severe complications were seen in one patient in group E and also in group F. CONCLUSIONS: The results suggest that 2-year intervals should be used between colonoscopies in patients with previous pedunculated adenomas and small, flat and sessile adenomas, whereas larger, flat and sessile adenomas may need intervals of 1 year. PMID- 16716976 TI - Efficacy of s-type stents for the treatment of the main pancreatic duct stricture in patients with chronic pancreatitis. AB - OBJECTIVE: Stents have been used to relieve pancreatic duct stricture and upstream dilatation. However, many of these stents are straight-type stents originally manufactured for biliary use. A plastic stent that was developed for use in the pancreatic duct was used in this study and its usefulness investigated. MATERIAL AND METHODS: The stent (s-type stent: 10 Fr in diameter) has two alternate flexions and the shape resembles the tilde mark " approximately " in appearance. After obtaining informed consent, stents were placed in 20 patients with abdominal pain caused by chronic pancreatitis and stricture of the distal main pancreatic duct. The stents were removed according to the clinical manifestations and replaced with new ones if the stricture persisted. RESULTS: In total, 33 stents were placed in 20 patients. Pain relief was attained in 19 patients (95%). The stricture improved after one stenting in 8 patients (40%). Owing to persistent stricture, the stenting was repeated in 11 patients. The 50% stent indwelling period was 369.0 days. No proximal or distal migration of the stent occurred and there were no serious complications. CONCLUSIONS: In view of its long durability as a stent and no migration, the s-stent is safe and useful for the management of pancreatic ductal strictures in patients with chronic pancreatitis. PMID- 16716977 TI - Treatment and outcome in pancreatic pseudocysts. AB - OBJECTIVE: Pancreatic pseudocyst is a common complication of both acute and chronic pancreatitis. The aim of the present study was to evaluate the efficacy and complications of different treatment regimens. MATERIAL AND METHODS: All patients > or =15 years of age admitted to Lund University Hospital from 1994 to 2003 with pancreatic pseudocysts were analysed retrospectively. Pseudocysts were defined according to the Atlanta classification. RESULTS: Forty-four patients (29 M (66%), mean age 55+/-14 years) were included in the study, and all were subjected to treatment on 88 occasions. Mean size of pseudocysts at diagnosis was 9.6+/-6.8 cm (1.5-40 cm). Recurrence after treatment was 1.0+/-1.1 times (range 0 4). No difference was found in recurrence rate or pseudocyst size when comparing conservative versus interventional treatment, but patient weight was higher (p=0.013) and acute pancreatitis was more frequent (p=0.046) in conservatively treated patients. Surgical treatment tended to be associated with a lower recurrence rate as compared with percutaneous treatments. The rate of hospital admissions was a median 3 (0-16) and median length of stay (LOS) was 12 days (0 141 days). Six patients (14%) had complications and 3 died (7%). Pseudocysts > or =8 cm did not differ significantly from smaller pseudocysts regarding the choice of conservative treatment, LOS, recurrence and gastrointestinal obstruction, but there was a trend towards more complications in the group with larger pseudocysts (5 versus 1). CONCLUSIONS: Patients with pancreatic pseudocysts require frequent hospital admissions and repeated treatments. Larger pseudocysts do not imply more recurrences. The lowest recurrence rate overall was seen after open surgery. PMID- 16716978 TI - Abnormal relationship between dissociation and hypnotic susceptibility in irritable bowel syndrome. PMID- 16716979 TI - Hyperalbuminemia and elevated transaminases associated with high-protein diet. AB - While high protein diets are increasing in popularity, there is a lack of data on their potential adverse effects. We describe two patients on high protein supplements and exercising for physical fitness. Both developed intermittent abdominal pain, transient elevations in transaminases and hyperalbuminemia without there being any identifiable cause. The symptoms and abnormalities on the laboratory tests resolved after the high protein intake was discontinued. While the pathogenesis and importance of these abnormalities need further study, the findings raise concerns regarding the safety of high protein diets combined with high intensity exercise. PMID- 16716980 TI - Granulation and fibrotic esophagus. AB - There is very little information in the medical literature on fibrotic esophageal lesions. These lesions do not appear to be malignant, but are recalcitrant to different treatments including removal, and the use of esophageal stents does not discourage the lesion from continuing to grow. The cause of these lesions remains unknown and the reasons for overgrowth in stents are poorly understood. We report on a patient who presented with dysphagia and a recalcitrant fibrotic esophageal lesion. PMID- 16716981 TI - Octreotide in the treatment of refractory ascites of cirrhosis. PMID- 16716982 TI - Metabolism and nutrition in patients with moderate and severe traumatic brain injury: A systematic review. AB - PRIMARY OBJECTIVE: To examine the evidence on the metabolic state and nutritional treatment of patients with moderate-to-severe traumatic brain injury (TBI). RESEARCH DESIGN: A systematic review of the literature. METHODS AND PROCEDURES: From 1547 citations, 232 articles were identified and retrieved for text screening. Thirty-six studies fulfilled the criteria and 30 were accepted for data extraction. MAIN OUTCOMES AND RESULTS: Variations in measurement methods and definitions of metabolic abnormalities hampered comparison of studies. However, consistent data demonstrated increased metabolic rate (96-160% of the predicted values), of hypercatabolism (-3 to -16 g N per day) and of upper gastrointestinal intolerance in the majority of the patients during the first 2 weeks after injury. Data also indicated a tendency towards less morbidity and mortality in early fed patients. CONCLUSIONS: The impact of timing, content and ways of administration of nutritional support on neurological outcome after TBI remains to be demonstrated. PMID- 16716983 TI - A personal intervention substance abuse treatment approach: Substance abuse treatment in a least restrictive residential model. AB - The following paper will detail demographic information about alcohol and substance abuse issues in the acquired brain injury (ABI) population. Included in such a review will be a systematic presentation of the current data on the incidence of such issues as well as a comprehensive treatment model with outcome data provided. Pre- and post-test measures of separate functional outcome areas were employed to assess the effects of the treatment package. The paper will document the core elements of a substance abuse treatment programme that is based upon community inclusion and positive, proactive treatment methods in facilitating decreases in substance abuse behaviour. The programme employed a host of individual and group session protocols specifically targeted at education and self-monitoring of behaviour in order to facilitate more effective self control of addictive behaviours. The positive outcomes achieved by the participants involved in the study provide clinical justification for a proactive behavioural approach to substance abuse in the ABI population. PMID- 16716984 TI - Domains of a TBI minimal data set: community reintegration phase. AB - Efforts to establish a minimal data set for the period of community integration after traumatic brain injury (TBI) have been the focus of a working group of leading researchers, clinicians and advocates attending the Galveston Brain Injury Symposium in 2001 and 2002. The purpose of the data set is to increase the consistency and interpretability of TBI research and programme evaluation by providing a common set of measures to document key aspects of community integration. It is intended to be sensitive to intervention, comprehensive, reflect satisfaction and outcome and promote future treatment/therapy. In deciding on the components of a minimal data set, the tri-partite model of outcomes was adopted (initially employed in psychotherapy, adapted to TBI) which focuses on the needs of the individual, society and the health provider. Scales are recommended for the respective components of the model. PMID- 16716985 TI - The structure of functional and community outcomes following traumatic brain injury. AB - OBJECTIVES: To investigate the dimensionality of functional and community outcomes following serious TBI. To identify items that fit, misfit or are redundant, as well as to assess person misfit. METHODS: Rating-scale (Rasch) analysis was applied to 1-year follow-up data from 231 cases in the US National TBI Model Systems database. Items selected for analysis included all items indicative of global outcomes, disability, activity or participation. RESULTS: A powerful singular measurement dimension was identified. Item reliability was very high (0.98), as was person reliability (0.97). The dimension fit over 90% of cases; that is approximately 10% of cases displayed anomalous patterns of functioning that indicated that their functioning was not measurable in terms of the general dimension identified. There was tension within the dimension between ratings of dependency (FIM) and cognitive functioning in everyday life (NFI). Most-but not all-neuropsychological test scores misfit the outcome dimension. CONCLUSIONS: Actual dimensionality was distinct from the named scales employed. A unidimensional measure model fit the data much better than expected. This outcome dimension might be called 'general community functioning'. In the future, it should be possible to develop more valid and parsimonious measures of community outcomes following TBI. PMID- 16716986 TI - MR imaging of head trauma: visibility of contusions and other intraparenchymal injuries in early and late stage. AB - The aim of this study was to investigate the visibility of traumatic brain lesions on conventional magnetic resonance images (MRI) in early and late phase. Thirty-six patients were studied 1 week and 1 year after a traumatic brain injury. A similar MRI technique was used in both studies; T2-weighted fast or turbo spin echo images, fluid attenuated inversion recovery (FLAIR) images and T1 weighted images were used for analysis. The number and extent of contusions and semi-quantitative score of other traumatic intraparenchymal lesions were compared in the early and late phase. Contusions were seen in 18 patients both in acute and 1 year MRI; the number and extent of visible contusions was significantly decreased at 1 year. Other traumatic intraparenchymal lesions were detected in 12 patients in early MRI and in 10 patients in late MRI. The number of visible lesions and the semi-quantitative scores were significantly lower at 1 year. There is a significant decrease in the visibility of both cortical contusions and other intraparenchymal injuries in late MRI studies compared with studies in acute stage using conventional imaging techniques. Thus, early phase MRI is essential for the detection of brain injury at least using conventional imaging techniques. PMID- 16716987 TI - Working memory and proverb comprehension in adolescents with traumatic brain injury: a preliminary investigation. AB - PRIMARY OBJECTIVE: This study investigated the relationship between working memory and comprehension of low-familiarity proverbs in adolescents with traumatic brain injury (TBI). METHODS AND PROCEDURES: Ten adolescents, aged 12-21 years who had suffered a TBI prior to the age of 10 years and 10 individually age matched peers with typical development participated in the study. The participants listened to short paragraphs containing a proverb and interpreted the meaning of the proverb using a forced-choice task. In addition, participants engaged in a task that evaluated working memory ability. MAIN OUTCOMES AND RESULTS: Analysis revealed that individuals with TBI differed from their non injured peers in their understanding of proverbs. In addition, working memory capacity influenced performance for all participants. CONCLUSIONS: The importance of considering working memory when evaluating figurative language comprehension in adolescents with TBI is highlighted. Implications for future research, particularly with regard to varying working memory and task demands, are considered. PMID- 16716989 TI - Dysautonomia and heart rate variability following severe traumatic brain injury. AB - PRIMARY OBJECTIVE: To investigate disconnection theories postulated as the cause of dysautonomia following severe traumatic brain injury (TBI) through analysis of heart rate variability (HRV). METHODS AND PROCEDURES: Data were collected on age matched subjects with and without dysautonomia (eight subjects in each group) and 16 non-injured controls. Data included injury details, continuous electrocardiograph recordings and rehabilitation outcome. MAIN OUTCOMES AND RESULTS: The TBI group revealed significant differences in HRV parameters both compared to controls and between dysautonomic and non-dysautonomic subjects. Additionally, HRV parameters for dysautonomic subjects showed evidence of an uncoupling of the normal relationship between heart rate and sympathetic/parasympathetic balance. HRV changes persisted for the dysautonomia group for a mean of 14 months post-injury. CONCLUSIONS: Dysautonomic subjects revealed prolonged uncoupling of heart rate and HRV parameters compared to non dysautonomic subjects and controls. These findings represent direct pathophysiological evidence supporting the disconnection theory postulated to produce dysautonomia following TBI. PMID- 16716988 TI - Social communication skills in persons with post-acute traumatic brain injury: three perspectives. AB - OBJECTIVE: To describe social communication skills problems identified by individuals with traumatic brain injury (TBI) compared to significant other (SO) and clinician ratings; and associations between these skills and participation outcome measures. DESIGN: Cohort study. METHODS: Sixty individuals with TBI > or = 1 year post-injury were administered measures of social communication, societal participation, social integration and life satisfaction. Clinicians and SOs rated the social communication skills of the subjects. RESULTS: Subjects were able to identify social communication skills problems, associated with lower ratings of community integration and satisfaction with life. Males reported higher scores in social communication and social integration than females. SOs and clinicians identified more social skills problems than subjects. CONCLUSIONS: Persons with TBI experience social communication skills deficits, associated with decreased societal participation and life satisfaction. Further research is needed to determine efficacy of social communication skills treatment and association with improved participation and satisfaction with life. PMID- 16716990 TI - The development of the therapeutic working alliance, patients' awareness and their compliance during the process of brain injury rehabilitation. AB - PURPOSE: To examine the development and interaction of the therapeutic alliance, patients' compliance and awareness during the process of brain injury rehabilitation and the role of demographic and injury related variables in this process. Subjects were 86 patients who underwent a holistic neuropsychological outpatient rehabilitation programme. Patients had suffered a traumatic brain injury (n = 27), a cerebrovascular accident (n = 49) or another neurological insult (n = 10). MEASURES: The therapeutic alliance between clients and their primary therapists, clients' awareness and their compliance were rated four times during the 14-week rehabilitation programme. The therapeutic alliance was rated by both clients and therapist using the Working Alliance Inventory (WAI), awareness and compliance were rated by the therapists. RESULTS: The development of the process measures over time is described in the article. Clients' and therapists' perspectives on their alliance tended to converge over time. Clients' experience of their emotional bond with their therapist added as much to the prediction of clients' awareness as the localization of their brain injury. Clients' awareness was related to their compliance and mediated the impact of the therapeutic alliance on their compliance. DISCUSSION: A good working alliance is the basis of successful rehabilitative work. The article discusses therapeutic implications of the results. PMID- 16716991 TI - UFOV performance and driving ability following traumatic brain injury. AB - PRIMARY OBJECTIVE: To investigate the relationship between performance on the Useful Field of View Test (UFOV) and driving performance following traumatic brain injury (TBI). PARTICIPANTS: Sixty people with TBI referred for driving evaluation. MEASURES: Useful Field of View Test, Global Rating Scale and Driver Assessment Scale. RESULTS: Subject performance diminished as the complexity of the UFOV sub-tests increased. There was a significant relationship between UFOV performance, particularly on the second sub-test, and on-road driving performance. Subject age and Trail Making Test, Part B were also predictive of driving performance. CONCLUSIONS: The UFOV can be used as a screening measure to determine readiness to participate in an on-road driving assessment. PMID- 16716993 TI - Utility of Glasgow Coma Scale-Extended in symptom prediction following mild traumatic brain injury. AB - STUDY OBJECTIVE: To examine the efficacy of the Glasgow Coma Scale-Extended (GCS E) for the prediction of symptoms commonly associated with mild traumatic brain injury (TBI). METHOD: Three hundred and sixty-one participants with a mild TBI were evaluated using the GCS-E and the Standardized Assessment of Concussion. A sub-group of 185 participants took part in a more extensive evaluation, which also included measures of depression and vestibular symptoms. All participants had a Glasgow Coma Scale score of 15, but experienced varying lengths of post traumatic amnesia (PTA) as measured by the GCS-E. RESULTS: Use of the GCS-E for assessment of PTA duration revealed that longer lengths of amnesia following mild TBI were associated with greater incidence of dizziness, depression and cognitive impairments during the first weeks after injury. CONCLUSION: Results suggest that the GCS-E is a useful tool for the prediction of symptoms associated with mild TBI. PMID- 16716992 TI - Predictive value of S-100B protein and neuron specific-enolase as markers of traumatic brain damage in clinical use. AB - OBJECTIVE: S-100B and NSE proteins are considered to be neurobiochemical markers for the brain damage. The aim of this study was to consider the diagnostic and prognostic validity of the initial serum levels of S-100B and NSE in clinical use. METHODS: Forty-five patients with traumatic brain injury were included in this prospective study. Neurologic examination and CCT-scan were performed. S 100B and NSE were analysed. Patients were divided in two groups depending on the severity of injury. RESULTS: The results showed a significant difference between the S-100B serum concentration and the two groups-minor head injuries and severe head injuries. A statistically significant correlation was observed between an increase of S-100B and NSE serum values and a cerebral pathological finding in CT scans. CONCLUSION: The clear correlation between S-100B and NSE serum concentrations and CCT findings does not validate both markers as an independent predictor of diagnosis and prognosis of brain injury. PMID- 16716994 TI - The impact of mild developmental learning difficulties on neuropsychological recovery from head trauma. AB - OBJECTIVES: To investigate the prevalence of mild developmental learning difficulties (MDLD) in a group of patients who had sustained head trauma, to determine the impact on cognitive and neurobehavioural recovery. METHODS: A case control design comprised 136 consecutive cases (55 MDLD; 81 control). Information regarding a history of MDLD was obtained during clinical interview, with neurobehavioural problems recorded using a checklist. Neuropsychological assessment and questionnaire measures of anxiety and depression were completed. RESULTS: Between group cognitive differences were only found on Digit Symbol (p < 0.01) and Trails B (p < 0.05) tests. The MDLD group reported a higher incidence of impulsive aggression and mood swings (both p < 0.01). Adjustment problems in the MDLD group were inferred from higher anxiety and depression ratings (both p < 0.05). CONCLUSIONS: The MDLD group did not appear to display a more generalized deficit in cognitive functions. However, neurobehavioural problems, plus the incidence and degree of affective changes, suggest greater psychological vulnerability than controls. PMID- 16716995 TI - Feedback of brain-imaging findings: effect on impaired awareness and mood in acquired brain injury. AB - PRIMARY OBJECTIVE: To investigate the utility of feedback of brain-imaging findings as an intervention for improving impaired awareness, depression and anxiety levels. RESEARCH DESIGN AND METHODS: A prospective within-subjects repeated measures design was used with an intervention sample of 17 adults with acquired brain injury. Baseline and post-intervention measures included self report and questionnaire-based self-awareness instruments and self-report questionnaires for depression and anxiety. Participants also completed a range of neuropsychological tests. INTERVENTION: The intervention consisted of a session where a Consultant Neurologist explained the findings of brain scans in terms of pathological findings and possible neurobehavioural outcome. RESULTS: Scores on measures of unawareness and two of three mood measures decreased significantly following the intervention. This improvement was maintained at 2-week follow-up. CONCLUSION: Individuals with an acquired brain injury may benefit from a feedback procedure where the findings of brain scans are presented. PMID- 16716996 TI - Traumatic brain injury among Australian prisoners: rates, recurrence and sequelae. AB - PRIMARY OBJECTIVE: To establish the prevalence, severity, recurrence and sequelae of past traumatic brain injury (TBI) among individuals recently received into custody. RESEARCH DESIGN: Cross-sectional random sample of men recently received into the New South Wales' (NSW) criminal justice system. PROCEDURES: Participants were screened for a history of TBI including the injury setting, severity, treatment and sequelae of up to five separate TBI episodes. OUTCOMES AND RESULTS: Of 200 study participants, 82% endorsed a history of at least one TBI of any severity and 65% a history of TBI with a loss of consciousness (LOC). Multiple past TBIs were common, as were ongoing sequelae. Treatment for the TBI was more common among those TBIs with a LOC compared with no LOC (66% vs. 39%). CONCLUSIONS: Among individuals entering the criminal justice system, past TBI is common and often associated with ongoing neuropsychiatric and social sequelae. Screening for TBI at the point of reception may be warranted to better understand and treat those with ongoing neuropsychiatric sequelae arising from the TBI. PMID- 16716997 TI - The incidence of visual perceptual impairment in patients with severe traumatic brain injury. AB - PRIMARY OBJECTIVES: To investigate the incidence of visual perceptual impairments in a sample of patients with traumatic brain injury (TBI) using the Occupational Therapy Adult Perceptual Screening Test (OT-APST), compare incidence rates to a normative sample and explore the relationship between the presence of visual perceptual impairment and the severity of cognitive and functional impairment following TBI. RESEARCH DESIGN: Cohort study using a convenience sample of patients with TBI and a normative sample. METHODS AND PROCEDURES: Thirty-one patients with severe TBI and 195 healthy people were compared on the OT-APST and measures of cognition and function. MAIN OUTCOMES AND RESULTS: The most common impairments in the TBI sample were unilateral neglect (45.2%) and impairments of body scheme (25.8%) and constructional skills (25.8%). Significantly higher incidences of agnosia, apraxia, unilateral neglect and impairments in body scheme and constructional skills were found in the TBI sample compared to the normative sample. No significant relationship was found between the presence of visual perceptual impairments and the level of cognitive and functional impairment after TBI. CONCLUSIONS: Visual perceptual changes are evident in patients with severe TBI when compared to a normative sample. Routine use of a screening tool such as the OT-APST may help identify visual perceptual impairments in these patients and the need for more detailed assessment. PMID- 16716998 TI - Confirmatory factor structure of the Center for Epidemiologic Studies-Depression scale (CES-D) in mild-to-moderate traumatic brain injury. AB - PRIMARY OBJECTIVE: The Center for Epidemiologic Studies Depression scale (CES-D) is a frequently-used self-report measure of depressive symptom severity. Brief depression screening measures can be important in the identification and prediction of depression following traumatic brain injury. The objective of this study was to investigate the validity of the CES-D in measuring depressive symptoms in patients with mild-to-moderate TBI as it has been rarely used in neurologically compromised populations. RESEARCH DESIGN: Inception cohort. METHODS AND PROCEDURES: The CES-D was administered to 340 participants with mild to-moderate TBI at 3-months post-injury. MAIN OUTCOMES AND RESULTS: Confirmatory factor analysis of the CES-D indicated that the data are a reasonable fit similar to that of Radloff 's original 4-factor model. CONCLUSIONS: These findings suggest that the CES-D may be appropriate for use in patients with mild-to moderate TBI. PMID- 16716999 TI - EMA assessment of tongue-jaw co-ordination during speech in dysarthria following traumatic brain injury. AB - PRIMARY OBJECTIVE: To investigate the spatio-timing aspects of tongue-jaw co ordination during speech in individuals with traumatic brain injury (TBI). It was hypothesized that both timing and spatial co-ordination would be affected by TBI. RESEARCH DESIGN: A group comparison design wherein Mann-Whitney U-tests were used to compare non-neurologically impaired individuals with individuals with TBI. METHODS AND PROCEDURES: Nine non-neurologically impaired adults and nine adults with TBI were involved in the study. Electromagnetic articulography (EMA) was used to track tongue and jaw movement during /t/ and /k/, embedded in sentence and syllable stimuli. MAIN OUTCOMES AND RESULTS: Analysis of group data did not reveal a significant difference in spatio-timing tongue-jaw co-ordination between the control group and TBI group. On an individual basis, a proportion of individuals with TBI differed from non-neurologically impaired participants with regard to articulatory order and percentage of jaw contribution to /t/. CONCLUSIONS: EMA assessment results supported perceptual data; those adults who presented with severe articulatory disturbances exhibited the most deviant spatio timing tongue-jaw co-ordination patterns. This finding could provide a new and specific direction for treatment, directed at combined movement patterns. PMID- 16717000 TI - Misconceptions about brain injury: a survey replication study. AB - PRIMARY OBJECTIVE: The researchers sought to update information about the public's knowledge about brain injury and to document progress made toward correcting misconceptions. RESEARCH DESIGN: Survey replication. METHODS AND PROCEDURES: Three hundred and eighteen respondents from the lay public answered questions about general brain injury knowledge, coma and unconsciousness, memory deficits and brain injury recovery. Chi-square analyses revealed significant differences among respondents based on age, gender, education and personal experience with brain injury or brain injury survivors. Findings were also compared with those reported by previous researchers administering similar questionnaires. MAIN OUTCOMES AND RESULTS: Misconceptions persist regarding some brain injury sequelae. Although most respondents knew general information, large numbers reported incorrect beliefs about memory problems, coma and unconsciousness and recovery. CONCLUSIONS: Despite the frequency with which it occurs, the general public persists in holding misconceptions about brain injury. Greater public awareness is necessary to prompt appropriate health care funding and rehabilitative service decisions. PMID- 16717002 TI - Patient safety. PMID- 16717003 TI - Patient safety during medication administration: the influence of organizational and individual variables on unsafe work practices and medication errors. AB - Medication errors are a leading cause of unintended harm to patients, both in Australia and internationally, and there is now a concerted attempt to identify and correct individual and workplace factors that encourage medication errors. The current study used structural equation modelling to measure organizational climate and to test a model with hypothesized links between climate and unsafe medication administration behaviours. The study also examined the possible mediating role of stress and morale. Data were collected from 176 nurses working in rural areas in Australia. The model provided a reasonable fit to the data with organizational climate accounting for 39% of the variance in individual distress, which in turn explained 7% of the variance in self-reported violations. The only variable that made a direct contribution to errors was violations, which accounted for 24% of the variance in medication errors. These findings highlight the importance of monitoring the state of the whole health system. Deficiencies at the organizational level affect the psychological well-being of hospital employees, and distressed employees are more likely to engage in substandard work practices that ultimately endanger the patients under their care. PMID- 16717004 TI - Work stress and patient safety: observer-rated work stressors as predictors of characteristics of safety-related events reported by young nurses. AB - This study investigates the link between workplace stress and the 'non singularity' of patient safety-related incidents in the hospital setting. Over a period of 2 working weeks 23 young nurses from 19 hospitals in Switzerland documented 314 daily stressful events using a self-observation method (pocket diaries); 62 events were related to patient safety. Familiarity of safety-related events and probability of recurrence, as indicators of non-singularity, were the dependent variables in multilevel regression analyses. Predictor variables were both situational (self-reported situational control, safety compliance) and chronic variables (job stressors such as time pressure, or concentration demands and job control). Chronic work characteristics were rated by trained observers. The most frequent safety-related stressful events included incomplete or incorrect documentation (40.3%), medication errors (near misses 21%), delays in delivery of patient care (9.7%), and violent patients (9.7%). Familiarity of events and probability of recurrence were significantly predicted by chronic job stressors and low job control in multilevel regression analyses. Job stressors and low job control were shown to be risk factors for patient safety. The results suggest that job redesign to enhance job control and decrease job stressors may be an important intervention to increase patient safety. PMID- 16717005 TI - Patient safety in outpatient surgery: the viewpoint of the healthcare providers. AB - The objective of this study is to understand the viewpoint of healthcare providers with regard to patient safety in outpatient surgery settings. Two methods were used to gather data from the healthcare providers: (1) questionnaire with open-ended questions about six predefined stages of the patient care process; (2) survey with closed questions. With the first method, the main quality and safety of care issues concerned communication to patients, coordination of reports and forms, patient and staff time pressures and standards of care. The first two stages of the outpatient surgery process, i.e. patient work-up prior to day of surgery and patient admission and preparation on the day of surgery, yielded many more comments than the other four stages. The results of the structured questionnaire show that, overall, the healthcare providers report high quality of care provided by themselves (98%) and their surgery centre (96%). With regard to patient safety (i.e. cancellations of surgeries, patient safety problems and serious mistakes), there was a clear difference in perceptions reported by the physicians vs. the nurses and other staff. Nurses and other staff were more likely to report patient safety problems than physicians. The combination of qualitative data from the initial questionnaire and the quantitative data from the structured questionnaire provides a rather complete view of the outpatient surgery staff perceptions of quality and safety of care. This research highlights the importance of getting input from the healthcare providers regarding the quality and safety of care rather than relying only on traditional measures about patient outcomes. PMID- 16717006 TI - From cognition to the system: developing a multilevel taxonomy of patient safety in general practice. AB - The paper describes the process of developing a taxonomy of patient safety in general practice. The methodologies employed included fieldwork, task analysis and confidential reporting of patient-safety events in five West Midlands practices. Reported events were traced back to their root causes and contributing factors. The resulting taxonomy is based on a theoretical model of human cognition, includes multiple levels of classification to reflect the chain of causation and considers affective and physiological influences on performance. Events are classified at three levels. At level one, the information-processing model of cognition is used to classify errors. At level two, immediate causes are identified, internal and external to the individual. At level three, more remote causal factors are classified as either 'work organization' or 'technical' with subcategories. The properties of the taxonomy (validity, reliability, comprehensiveness) as well as its usability and acceptability remain to be tested with potential users. PMID- 16717007 TI - Error recovery in a hospital pharmacy. AB - A field study was performed in a hospital pharmacy aimed at identifying positive and negative influences on the process of detection of and further recovery from initial errors or other failures, thus avoiding negative consequences. Confidential reports and follow-up interviews provided data on 31 near-miss incidents involving such recovery processes. Analysis revealed that organizational culture with regard to following procedures needed reinforcement, that some procedures could be improved, that building in extra checks was worthwhile and that supporting unplanned recovery was essential for problems not covered by procedures. Guidance is given on how performance in recovery could be measured. A case is made for supporting recovery as an addition to prevention based safety methods. PMID- 16717008 TI - Error detection: a study in anaesthesia. AB - Although error has been shown as the main cause of accidents in complex systems, little attention has been paid to error detection. However, reducing the consequences of error depends largely on error detection. The goal of this paper is to synthesize the existing scientific knowledge on error detection, mostly based on studies conducted in laboratory or self reporting and to further knowledge through the analysis of a corpus of cases collected in a complex system, anaesthesia. By doing this, this paper is better able to describe how this knowledge can be used to improve understanding of error detection modes. An anaesthesia accident reporting system developed and organized at two Belgian University Hospitals was used in order to collect information about the error detection patterns. Results show that detection of errors principally occurred through the standard check (routine monitoring of the environment). Significant relationships were found between the type of error and the error detection mode, and between the type of error and the training level of the anaesthetist who committed the error. PMID- 16717009 TI - Prospective memory failures as an unexplored threat to patient safety: results from a pilot study using patient simulators to investigate the missed execution of intentions. AB - This study investigated failures of prospective memory (PM) as a relevant but neglected error type in medicine. A patient simulator was used to investigate PM failures. The influence of subjective importance (high, low) and type of intention (educational, internal, external) on the (missed) execution of intention was investigated in a 2 x 2 design. The effects on missed executions by importance (high < low) and type of intention (educational < external < internal) were hypothesized. Of 73 valid intentions in 40 prepared simulator scenarios 19 (26%) were missed overall. A total of 64% of unimportant and 80% of important intentions were executed 79% of educational 67% of external and 72% of internal intentions were executed. Neither difference was statistically significant using chi(2) tests. Interaction was significant for missed executions (p = 0.025; n = 19; df = 2; chi(2) = 7.41) and for executions (p = 0.002; n = 54; df = 2; chi(2) = 12.50). Despite low statistical support and some methodological limitations, it was possible to show that PM failures are relevant to patient safety and that patient simulators are a suitable but so far unused tool for their investigation. PMID- 16717010 TI - Identification of systems failures in successful paediatric cardiac surgery. AB - Patient safety will benefit from an approach to human error that examines systemic causes, rather than blames individuals. This study describes a direct observation methodology, based on a threat and error model, prospectively to identify types and sources of systems failures in paediatric cardiac surgery. Of substantive interest were the range, frequency and types of failures that could be identified and whether minor failures could accumulate to form more serious events, as has been the case in other industries. Check lists, notes and video recordings were employed to observe 24 successful operations. A total of 366 failures were recorded. Coordination and communication problems, equipment problems, a relaxed safety culture, patient-related problems and perfusion related problems were most frequent, with a smaller number of skill, knowledge and decision-making failures. Longer and more risky operations were likely to generate a greater number of minor failures than shorter and lower risk operations, and in seven higher-risk cases frequently occurring minor failures accumulated to threaten the safety of the patient. Non-technical errors were more prevalent than technical errors and task threats were the most prevalent systemic source of error. Adverse events in surgery are likely to be associated with a number of recurring and prospectively identifiable errors. These may be co incident and cumulative human errors predisposed by threats embedded in the system, rather than due to individual incompetence or negligence. Prospectively identifying and reducing these recurrent failures would lead to improved surgical standards and enhanced patient safety. PMID- 16717011 TI - Measuring intra-operative interference from distraction and interruption observed in the operating theatre. AB - An observational tool was developed to record distraction and interruption in the operating theatre during surgery. Observed events were assigned to pre-defined categories and rated in relation to the level of team involvement - the sum of which was treated as a measure of intra-operative interference. Many events (0.29 +/- 0.02 per min) were observed and rated in 50 general operations sampled from a single operating theatre. The rating of individual events (r(s) = 0.65) and of cases (r(s) = 0.89) correlated between independent observers. Interference levels (1.04 +/- 0.07/min) also correlated with door opening frequency (0.68 +/- 0.03/min) (r = 0.47, p < 0.001). Some sources of interference were intrinsic to the work of the surgical team, including equipment, procedure and environment, while others were extraneous, including bleepers, phone calls and external staff. The findings highlight the need to further develop measures of interference, to assess its variation, intensity and its effect on surgical team performance. PMID- 16717012 TI - A review of environmental hazards associated with in-patient falls. AB - Slips, trips and falls present the greatest risk to in-patients in terms of exposure (frequency of occurrence) but only present a low severity risk in terms of mortality. The risk factors have been categorized as intrinsic (individual to the patient, e.g. visual impairment, balance problems and medicine use) or extrinsic (environmental). Many recommendations have been made concerning the management of environmental hazards but, of these, only beds rails have supporting research evidence. Other recommendations include patient assessment, footwear, flooring, lighting, staffing levels and bed alarms. However, three systematic reviews and the current narrative review have all failed to find research evaluating the benefits of these recommendations. The most robust evidence relates to the use of bed rails. This research suggests that bed rails not only fail to reduce the frequency of falls, but may also exacerbate the severity of injury. As Maslow's Hierarchy of Needs model has been used as a framework for nursing models of care, it was chosen as the basis for the development of an environmental hazard assessment model. The environmental hazards are revisited using this model in order to take an ergonomic or patient centred approach for risk assessment. PMID- 16717013 TI - Considering the impact of medicine label design characteristics on patient safety. AB - Medication errors involving patients receiving the wrong medicines, the wrong dosages or failure to take medicines according to the prescribed schedule are a substantial threat to patient safety. In the medical domain, research evidence on the benefits of improved labelling are piecemeal and often single-product or single-manufacturer driven and often do not inform the more general process of label design. Government and other guidelines on this topic are often low level and non-specific, often failing to give evidence-based guidance. However, there is a wealth of evidence-based research findings in related areas such as food labelling, chemical labelling and more general warnings research, which can provide systematic evidence on the effects of design characteristics such as font size, colour, signal words and linguistic usage on crucial performance variables such as compliance, understandability and discriminability. This research is reviewed and its relevance to medicine labelling is presented. PMID- 16717015 TI - Audiology in South Africa. AB - Audiology in South Africa is an established profession facing the challenge of serving a diverse population in predominantly developing contexts. The profession has developed over the last half century from an adjunct to speech-language pathology into a profession in its own right. Several tertiary institutions offer undergraduate training in audiology with optional postgraduate qualifications. Institutions are continually adapting to a profession characterised by rapid change--evidenced even in the very composition of the profession itself. This article aims to provide an overview of the development and current status of audiology as a profession in South Africa. PMID- 16717016 TI - The suffering of tinnitus in childhood and adolescence. AB - OBJECTIVES: Investigate the profile of young patients complaining of tinnitus and study the prevalence and the severity of tinnitus in schoolchildren. DESIGN: Ninety-five consecutive patients (55 boys), 8-20 years old, seeking help for tinnitus at our clinic were studied using audiometry, structural interviews, Tinnitus Severity Questionnaire (TSQ) and Hospital Anxiety and Depression Scale (HAD), together with Visual Analog Scales (VAS) on tinnitus loudness and annoyance. RESULTS: The onset of tinnitus was twice as often sudden than gradual. In 54% of the children, tinnitus had started after noise exposure, most commonly listening to music. Correlations were found between the pure-tone average (PTA) of 3, 4, and 6 kHz and TSQ scores; between TSQ and HAD subscales; and between TSQ and VAS. According to the Hospital Anxiety Depression Scale, 32% and 14.5%, respectively, were above the cut level for clinical anxiety and depressive disorders. The girls showed more signs of anxiety disorders than the boys. CONCLUSIONS: The majority of the help seekers had been exposed to noise, mostly music. Predisposing factors for tinnitus severity are high-frequency hearing loss and anxiety and depressive disorders. PMID- 16717017 TI - Reported hearing protection use in young adults from Sweden and the USA: effects of attitude and gender. AB - The present study investigates differences between a Swedish and an American sample of young students regarding attitudes towards noise and the use of hearing protection at concerts. The study population was comprised of 179 participants from Sweden and 203 participants from the United States, who ranged in age from 17 to 21 years. Questionnaires were used to gather information on hearing symptoms and attitudes towards noise (Youth Attitude to Noise Scale). Multivariate analysis of variance revealed that attitudes towards noise differed significantly due to gender and country. Men had slightly more positive attitude towards noise than women, and men from the USA had more positive attitudes than men from Sweden. Least positive were the women from Sweden (except regarding attitudes towards the ability to concentrate in noisy environments). Multivariate logistic regression analysis was used to examine the influence of attitudes towards noise and country on young people's use of hearing protection at concerts. The results indicated that attitudes and country explained 50% of the variance in use of hearing protection. PMID- 16717018 TI - The ASSR: clinical application in normal-hearing and hearing-impaired infants and adults, comparison with the click-evoked ABR and pure-tone audiometry. AB - The objective of this study was to investigate the clinical application of the ASSR (GSI Audera). It was completed in two parts: Study 1. Correlation between the ASSR-based threshold estimations and the conventional pure-tone thresholds in adults; and Study 2. Correlation between the average of the 2-4 kHz ASSR-based threshold estimations and c-ABR thresholds in children. The ASSRs were recorded in awake adults and sleeping infants with a range of hearing loss at CFs of 0.5 to 4 kHz and MFs between 46 and 95 Hz. The results show that in hearing-impaired adults (thresholds > 40 dBHL) good correlations can be observed between the behavioural thresholds and the ASSR-based threshold estimations. For the normal- to near-normal-hearing adults, a significant correspondence exists between the ASSR-based threshold estimations and FPTA. In children, strong correlations were found between the c-ABR and the 2-4 kHz ASSR-based threshold estimation average. These studies illustrate that the GSI Audera ASSR can accurately predict the behavioural audiogram in hearing-impaired subjects. In subjects with normal hearing the individual ASSR-based threshold estimations scatter too much. Instead the average of the ASSR-based threshold estimations corresponds well with the FPTA. PMID- 16717019 TI - Spectral loudness summation as a function of duration for hearing-impaired listeners. AB - Spectral loudness summation was measured for 10- and 1000-ms long bandpass-noise signals with bandwidths in the range of 0.2 to 6.4 kHz centered at 2 kHz for nine listeners with hearing impairment of primarily cochlear origin. Loudness matches between signals with the same duration and different bandwidth were obtained using an adaptive two-interval, two-alternative forced-choice procedure with interleaved tracks. The 3200-Hz wide reference signal had a level of 45 dB or 65 dB SPL. Every signal was individually frequency equalized prior to presentation in order to ensure audibility of all spectral components for the two reference input levels. Generally the same amount of spectral loudness summation for 10- and 1000-ms long signals was obtained for both reference levels. However, some hearing-impaired listeners show, similarly to normal-hearing listeners, a larger spectral loudness summation for short than for long duration signals, indicating that duration effects in spectral loudness summation are caused by retrocochlear processes. PMID- 16717020 TI - Evaluation of an audiovisual-FM system: speechreading performance as a function of distance. AB - A research program was undertaken to evaluate the efficacy of an audiovisual-FM system as a speechreading aid. The present study investigated the effects of the distance between the talker and the speechreader on a visual-speech perception task. Sentences were recorded simultaneously with a conventional Hi8 mm video camera, and with the microcamera of an audiovisual-FM system. The recordings were obtained from two talkers at three different distances: 1.83 m, 3.66 m, and 7.32 m. Sixteen subjects completed a visual-keyword recognition task. The main results of the investigation were as follows: For the recordings obtained with the conventional video camera, there was a significant decrease in speechreading performance as the distance between the talker and the camera increased. For the recordings obtained with the microcamera of the audiovisual-FM system, there were no differences in speechreading as a function of the test distances. The findings of the investigation confirm that in a classroom setting the use of an audiovisual-FM system may constitute an effective way of overcoming the deleterious effects of distance on speechreading performance. PMID- 16717021 TI - Effects of applied contact force and volume control setting on output force levels of the BAHA Softband. AB - The BAHA Softband has been developed to provide a transcutaneous anchor for a BAHA until a child is a surgical candidate for the percutaneous BAHA implant. We tested the objective output force level of the BAHA Classic 300 and Compact connected to a Softband on an artificial mastoid to determine: (1) the effects of direct contact force on output force levels (dB); and (2) the required volume control setting to ensure audibility of speech (assuming an average adult reference equivalent threshold force level). Direct contact force was varied from 2 to 5 N in 1 N steps. Output force level increased with increasing contact force. However, the average increase was 3 dB or lower, suggesting that the contact force is of minor importance. Volume control setting appears to be of much greater importance. It is suggested that the volume setting of either device be set to at least 2.5 to ensure audibility of conversational speech. Data from normal-hearing adults with simulated conductive hearing losses are presented to validate this conclusion. PMID- 16717022 TI - Hearing loss induced by occupational and impulse noise: results on threshold shifts by frequencies, age and gender from the Nord-Trondelag Hearing Loss Study. AB - The aim of the study was to compare the frequency-specific effects of noise on hearing acuity across the range 250-8000 Hz and the extent to which the patterns of frequency-specific threshold shifts differ between occupational noise and impulse noise. Pure-tone audiometry was administered to an adult general population sample with 51 975 subjects who also provided questionnaire information about noise exposure and other risk factors. Threshold shifts induced by life-long occupational noise and impulse noise (mostly shooting) were estimated separately in six age and sex groups for eight frequencies. Reported noise exposure, as well as observed threshold shifts, were moderate among women. Threshold shifts averaged over both ears among subjects in the higher 2% of exposure to occupational noise, reached 13 dB (3000 Hz, age 65 years + ) among men and were generally largest at 3000-4000 Hz. The shifts induced by impulse noise reached approximately 8 dB among men 45-64 years and men 65 years+. The effects of impulse noise were strongest at 3000-8000 Hz and varied little within this frequency range. PMID- 16717023 TI - Chlorosilane acute inhalation toxicity and development of an LC50 prediction model. AB - The acute inhalation toxicity of 10 chlorosilanes was investigated in Fischer 344 rats using a 1-h whole-body vapor inhalation exposure and a 14-day recovery period. The median lethal concentration (LC50(1)) for each material was calculated from the nominal exposure concentrations and mortality. Experimentally derived LC50(1) values for monochlorosilanes (4257-4478 ppm) were greater than those for dichlorosilanes (1785-2092 ppm), which were greater than those for trichlorosilanes (1257-1611 ppm). Apparent was a strong structure-activity relationship (r2 = .97) between chlorine content and LC50(1) value. Estimated LC50(1) values for mono-, di-, and trichlorosilanes were determined to be 3262, 1639, and 1066 ppm, respectively, utilizing this relationship and the lower limit of the 95% prediction interval. The LC50(1) values determined in this series of studies were greater than that reported for hydrogen chloride (3124 ppm), when expressed on a chlorine equivalence basis (3570-5248 ppm), demonstrating that the acute toxicity of these chlorosilanes is similar to or less than that for hydrogen chloride. The good correlation between chlorine content and LC50(1) provides a sound basis for estimation of LC50(1) for chlorosilanes not already evaluated. The use of structure-activity relationships is consistent with the chemical industry and federal agency initiatives to reduce, refine, and/or replace the use of animals in testing without compromising the quality of health and safety assessments. PMID- 16717024 TI - Health effects of subchronic exposure to environmental levels of hardwood smoke. AB - Hardwood smoke is a contributor to both ambient and indoor air pollution. As part of a general health assessment of multiple anthropogenic source emissions conducted by the National Environmental Respiratory Center, a series of health assays was conducted on rodents exposed to environmentally relevant levels of hardwood smoke. This article summarizes the study design and exposures, and reports findings on general indicators of toxicity, bacterial clearance, cardiac function, and carcinogenic potential. Hardwood smoke was generated from an uncertified wood stove, burning wood of mixed oak species. Animals were exposed to clean air (control) or dilutions of whole emissions based on particulate (30, 100, 300, and 1000 micromg/m3). F344 rats, SHR rats, strain A/J mice, and C57BL/6 mice were exposed by whole-body inhalation 6 h/day, 7 days/wk, for either 1 wk or 6 mo. Effects of exposure on general indicators of toxicity, bacterial clearance, cardiac function, and carcinogenic potential were mild. Exposure-related effects included increases in platelets and decreases in blood urea nitrogen and serum alanine aminotransferase. Several other responses met screening criteria for significant exposure effects but were not consistent between genders or exposure times and were not corroborated by related parameters. Pulmonary histopathology revealed very little accumulation of hardwood smoke particulate matter. Parallel studies demonstrated mild exposure effects on bronchoalveolar lavage parameters and in a mouse model of asthma. In summary, the results reported here show few and only modest health hazards from short-term to subchronic exposures to realistic concentrations of hardwood smoke. PMID- 16717025 TI - Vehicular air pollution, playgrounds, and youth athletic fields. AB - In spite of epidemiological evidence concerning vehicular air pollution and adverse respiratory/cardiovascular health, many athletic fields and school playgrounds are adjacent to high traffic roadways and could present long-term health risks for exercising children and young adults. Particulate matter (PM(1),0.02-1.0 microm diameter) number counts were taken serially at four elementary school athletic/playground fields and at one university soccer field. Elementary school PM1 measurements were taken over 17 days; measurements at the university soccer field were taken over 62 days. The high-traffic-location elementary school field demonstrated higher 17-day [PM1] than the moderate and 2 low traffic elementary school fields (48,890 +/- 34,260, 16,730 +/- 10,550, 11,960 +/- 6680, 10,030 +/- 6280, respective mean counts; p < .05). The 62-day mean PM1 values at the university soccer field ranged from 115,000 to 134,000 particles cm(-3). Lowest mean values were recorded at measurement sites furthest from the highway (approximately 34,000 particles cm(-3)) and followed a second order logarithmic decay (R2 = .999) with distance away from the highway. Mean NO2 and SO2 levels were below 100 ppb, mean CO was 0.33 +/- 1.87 ppm, and mean O3 was 106 +/- 47 ppb. Ozone increased with rising temperature and was highest in the warmer afternoon hours (R = .61). Although the consequence of daily recess play and athletic activities by school children and young athletes in high ambient [PM1] conditions has not yet been clearly defined, this study is a critical component to evaluating functional effects of chronic combustion-derived PM exposure on these exercising schoolchildren and young adults. Future studies should examine threshold limits and mechanistic actions of real-world particle exposure. PMID- 16717026 TI - Air pollution and hospital admissions for asthma in a tropical city: Kaohsiung, Taiwan. AB - This study was undertaken to determine whether there is an association between air pollutants levels and hospital admissions for asthma in Kaohsiung, Taiwan. Hospital admissions for asthma and ambient air pollution data for Kaohsiung were obtained for the period from 1996 through 2003. The relative risk of hospital admission was estimated using a case-crossover approach, controlling for weather variables, day of the week, seasonality, and long-term time trends. In the single pollutant models, on warm days (> or = 25 degrees C) statistically significant positive associations were found in all pollutants except sulfur dioxide (SO2). On cool days (< or = 25 degrees C) all pollutants were significantly associated with asthma admissions For the two pollutant models, CO and O3 were significant in combination with each of the other four pollutants on warm days. On cool days NO2 remained statistically significant in all the two-pollutant models. This study provides evidence that higher levels of ambient pollutants increase the risk of hospital admissions for asthma. PMID- 16717028 TI - Effects of environmental tobacco smoke on respiratory symptoms and pulmonary function. AB - The aim of this study was to examine the effects of environmental tobacco smoke (ETS) on pulmonary function and respiratory symptoms. During periodic medical examination, 392 French nonsmokers responded to an interviewer-administered questionnaire. Then spirometry was performed to assess pulmonary function. All of the subjects were carefully examined by two occupational physicians. ETS exposure at the workplace was more common than this exposure at home (20% vs. 5%). ETS exposure was significantly associated with forced vital capacity (FVC; -3.16%; 95% CI: -5.67 to -0.64) and forced expiratory volume in 1s (FEV1; -2.90%; 95% CI: -5.59 to -0.23). Abnormal FVC results were significantly increased in exposed subgroup [odds ratio = 2.71 (95% CI: 1.09 to 6.75)]. We did not find any significant dose-response relationship between ETS exposure and lung function results. The effects of ETS exposure on respiratory symptoms and diseases (asthma, wheezing, chronic bronchitis, and dyspnea) were not significant. Thus, this study showed that there was a significant inverse association between exposure to ETS and pulmonary function. Even pulmonary function results inferior to the lower limit of normal may be possible. A stricter legislation against ETS is proposed. PMID- 16717027 TI - Gene expression profiling in lung tissues from mice exposed to cigarette smoke, lipopolysaccharide, or smoke plus lipopolysaccharide by inhalation. AB - The purpose of this study was to investigate whether coexposure to lipopolysacchride (LPS) will heighten the inflammatory response and other pulmonary lesions in mice exposed to cigarette smoke, and thus to evaluate the potential use of this LPS-compromised mouse model as a model for chronic obstructive pulmonary disease (COPD) investigation. AKR/J male mice were exposed to HEPA-filtered air (sham control group), cigarette smoke (smoke group), LPS (LPS group), or smoke plus LPS (smoke-LPS group) by nose-only inhalation. Lungs were collected at the end of the 3-wk exposure and processed for microarray analysis. Clustering and network analysis showed decreased heat-shock response and chaperone activity, increased immune and inflammatory response, and increased mitosis in all three exposed groups. Two networks/function modules were exclusively found in the smoke-LPS group, that is, the downregulated muscle development/muscle contraction process and the upregulated reactive oxygen species production process. Notably, the number of genes and function modules/networks associated with inflammation was reduced in the smoke-LPS group compared to the LPS group. The most upregulated gene in the smoke group, MMP12, is a matrix metalloproteinase that preferentially degrades elastin and has been implicated in COPD development. NOXO1, which was upregulated in all three treatment groups, positively regulates the expression of a subunit of NADPH oxidase (NOX1), a major source of reactive oxygen species, and may play an important role in the pathogenesis of COPD. Serum amyloid A1, which is an acute phase systemic inflammation marker and can be induced by LPS exposure, was significantly upregulated in the LPS and smoke-LPS groups. MARCO, a scavenger receptor expressed in macrophages that may play a significant role in LPS-induced inflammatory response, was upregulated in the LPS group and the smoke-LPS group, but not in the smoke group. In conclusion, gene expression profiling identified genes and function modules that may be related to COPD pathogenesis and may be useful as biomarkers to monitor COPD progression. In addition, an LPS-compromised mouse model showed potential as a useful tool for studying cigarette smoke associated COPD. PMID- 16717029 TI - Short-term effects of smoking cessation on blood antioxidant parameters and paraoxonase activity in healthy asymptomatic long-term cigarette smokers. AB - The aim of this study was to compare short-term effects of smoking cessation on blood oxidant/antioxidant status, cholesterol levels, and paraoxonase activity. Sixteen healthy, asymptomatic long-term cigarette smokers (mean age: 35 +/- 9 yr) participated in the study in the smoking cessation program. After and before smoking cessation, subjects were examined for oxidant/antioxidant status, cholesterol level, paraoxonase activity, breath carbon monoxide levels, and blood carboxyhemoglobin values. When compared to previous values, subjects were revealed statistically significant decreases in malondialdehyde and carbon monoxide levels 4 wk after smoking cessation. The ratio of high-density lipoprotein (HDL) low-density lipoprotein (LDL) cholesterol was found to be increased. Significantly increased to paraoxonase activity was also observed in the blood samples obtained after cigarette cessation period. It was concluded that all these changes observed after smoking cessation might be of importance in the reduction of cardiovascular risk parameters in the smokers. PMID- 16717030 TI - Effect of sulfur dioxide inhalation on CYP2B1/2 and CYP2E1 in rat liver and lung. AB - Sulfur dioxide (SO2) is a ubiquitous air pollutant, present in low concentrations in the urban air and in higher concentrations in the working environment. In this study, we investigated the effects of inhaled SO2 on the O-dealkylase of pentoxyresorufin (PROD) and p-nitrophenol hydroxylases (p-NP) activities and mRNA levels of CYP2B1/2 and CYP2E1 in the lung and liver of Wistar rats. Male Wistar rats were housed in exposure chambers and treated with 14.11 +/- 1.53, 28.36 +/- 2.12, and 56.25 +/- 4.28 mg/m3 SO2 for 6 h/day for 7 days, while control rats were exposed to filtered air in the same condition. The mRNAs of CYP2B1/2 and 2E1 were analyzed in livers and lungs by using reverse-transcription polymerase chain reaction (RT-PCR). Our results showed that the PROD activities and mRNA of CYP2B1/2 were decreased in livers and lungs of rats exposed to SO2. The p-NP activities and mRNA of CYP2E1 were decreased in lungs but not in livers of rats exposed to SO2. Total liver microsomal cytochrome P-450 (CYP) contents were diminished in SO2-exposed rats. These results lead to two conclusions: (1) SO2 exposure can suppress CYP2B1/2 and CYP2E1 in lungs and CYP2B1/2 in livers of rats, thus modifying the liver and lung toxication/detoxication potential, and (2) the total liver microsomal CYP contents were diminished, although the activity and mRNA expression of CYP2E1 in rat livers were not affected by SO2 exposure. PMID- 16717031 TI - Hair as a biological indicator of drug use, drug abuse or chronic exposure to environmental toxicants. AB - In recent years hair has become a fundamental biological specimen, alternative to the usual samples blood and urine, for drug testing in the fields of forensic toxicology, clinical toxicology and clinical chemistry. Moreover, hair-testing is now extensively used in workplace testing, as well as, on legal cases, historical research etc. This article reviews methodological and practical issues related to the application of hair as a biological indicator of drug use/abuse or of chronic exposure to environmental toxicants. Hair structure and the mechanisms of drug incorporation into it are commented. The usual preparation and extraction methods as well as the analytical techniques of hair samples are presented and commented on. The outcomes of hair analysis have been reviewed for the following categories: drugs of abuse (opiates, cocaine and related, amphetamines, cannabinoids), benzodiazepines, prescribed drugs, pesticides and organic pollutants, doping agents and other drugs or substances. Finally, the specific purpose of the hair testing is discussed along with the interpretation of hair analysis results regarding the limitations of the applied procedures. PMID- 16717032 TI - Observations of alanine aminotransferase and aspartate aminotransferase in THRIVE studies treated orally with ximelagatran. AB - Treatment of acute venous thromboembolism (VTE) and prophylaxis of recurrent events has been investigated in the THRIVE (THRombin Inhibitor in Venous Thrombe Embolism) Treatment and the THRIVE III trial using the oral direct thrombin inhibitor ximelagatran. Alanine aminotransferase (ALAT) increased in 9.6% and 6.4% of patients in the THRIVE Treatment and THRIVE III trials, respectively. The authors analysed the time course of the ALAT and in additionally of aspartate aminotransferase (ASAT) in blood from 52 and 23 patients participating in the THRIVE Treatment and the THRIVE III trials in Germany. Analysis of variance for repeated measures and t test were performed. In the THRIVE Treatment trial, ALAT was significantly higher at week 2 for enoxaparin/warfarin (p => .0039, t test) and at months 3 and 6 for ximelagatran (p = .0453, p = .0014, respectively). ASAT and ASAT/ALAT ratio values did not increase and not differ for both groups. In the THRIVE III trial, ALAT and ASAT did not increase and did not differ compared to the comparator placebo. 2 x 36 mg Ximelagatran, induced higher ALAT values at months 3 and 6 compared to 2 x 24 mg ximelagatran (p = .0105, p = .0063, respectively). ASAT did not differ between the two doses of ximelagatran. The ASAT/ALAT ratios were lower at week 2 for enoxaparin/warfarin (t-test, p = .0032) and at month 3 and 6 for 2 x 36 mg versus warfarin or 2 x 24 mg Ximelagatran (p between .0187 and .0002). The authors conclude that ALAT increases dose dependently during therapy with ximelagatran. The less frequent and lower increase of ASAT values compared to ALAT values indicates a nontoxic effect of ximelagatran on liver cells. PMID- 16717033 TI - Acute toxicity pilot evaluation of proliferol in rats and swine. AB - Proliferol is an investigational new drug containing lidocaine hydrochloride 0.25%, dextrose 12.5%, glycerin 12.5%, and phenol 1.0% in aqueous solution. Despite extensive previous experience with similar drug solutions administered in humans by intraligamentous injection for chronic musculoskeletal conditions for over 50 years, animal toxicity data are unavailable. A pilot study was conducted to assess acute toxic effects prior to undertaking further assessment of this drug. Test animals were four Sprague-Dawley rats and four Yucatan mini-swine. Rats received injections into lumbar paraspinal muscles, whereas swine received injections into lumbosacral ligaments in an attempt to mirror the method of administration in humans. Two doses were studied equivalent to 1x and 5x the typical human dose. Outcomes measured at 24 h and 14 days included clinical observations, clinical chemistry, hematology, urinalysis, local tolerance, and major organ histopathology. In rats and swine, results from clinical chemistry, hematology, and urinalysis were indicative of acute local inflammation. At the high dose, marked (rats) and moderate (swine) short-term above-normal levels in certain liver enzymes were noted. In rats and swine, local tolerance results were indicative of acute local inflammatory changes in the skin, subcutis, and muscle around the injection sites. In rats and swine, major organ histopathology results did not reveal lesions attributable to the drug and clinical observations were within normal limits. In swine, fibroplasia was noted in deeper muscle tissues after 14 days. Injections of Proliferol in lumbar paraspinal muscles in rats and lumbosacral ligaments in swine elicited a modest acute local inflammatory response with no other indications of local or systemic toxicity. PMID- 16717034 TI - The effects of vehicles on the human dermal irritation potentials of allyl esters. AB - Allyl esters, frequently used in the fragrance industry, often contain a certain percentage of free allyl alcohol. Allyl alcohol is known to have a potential for delayed skin irritation. Also present in the finished product are different solvent systems, or vehicles, which are used to deliver the fragrances based upon their intended application. This study was conducted to determine whether different vehicles affect the skin irritation potential of five different allyl esters. The allyl esters tested were allyl amyl glycolate, allyl caproate, allyl (cyclohexyloxy)acetate, allyl cyclohexylpropionate, and allyl phenoxyacetate in the vehicles diethyl phthalate, 3:1 diethyl phthalate:ethanol, and 1:3 diethyl phthalate:ethanol at concentrations of 0.1%, 0.5%, 1.0%, and 2.0% (w/w). A modified cumulative irritation test was conducted in 129 human subjects. Test materials (0.3 ml) were applied under occlusion to skin sites on the back for 1 day (24 h) using Hill Top chambers. Irritation was assessed at 1, 2, 4, and 5 days following application of test materials. Cumulative irritation scores varied considerably among test materials. There were no delayed irritation observations. The highest irritation scores were observed at the 2.0% concentration for all test materials. The irritation scores for allyl amyl glycolate, allyl (cyclohexyloxy)acetate, and allyl phenoxyacetate were highest in 1:3 diethyl phthalate:ethanol, thus the resulting calculated no-observed-effect levels, 0.12%, 0.03%, and 0%, respectively, were much lower for this vehicle compared to the diethyl phthalate vehicle, 0.33%, 0.26%, 0.25%, respectively. These data showed a trend for lower concentration thresholds to induce irritation when higher levels of ethanol were used in the vehicle. PMID- 16717035 TI - Lead absorption and excretion in rats given insoluble salts of pectin and alginate. AB - Exposure to environmental lead remains a widespread problem in most industrialized countries. Usage of modern agents purposed for elimination of heavy metals as well as for therapy and prevention of chronic poisoning does frequently result in toxic signs. Dietary nonstarch polysaccharides were suggested to be effective when used for this purpose. The present study was conducted to estimate metal binding capacity and effects of calcium salts of pectate and alginate on lead absorption, distribution, and removal with feces. Under in vitro conditions calcium alginate showed the highest lead-binding capacity in comparison with other agents studied. Metal binding capacity of calcium pectate was slightly lower. In rats simultaneous administration of lead acetate and suspensions containing calcium alginate or calcium pectate prevented metal absorption and significantly reduced lead accumulation in inner organs and femur. In experiments estimating lead removal from inner organs and femur in rats preliminary exposed to the heavy metal, calcium alginate and calcium pectate were the most effective agents studied in comparison with others, as indicated by reduced lead concentration in organs and femur as well as increased metal content in feces of laboratory animals. The results suggest that calcium pectate and calcium alginate may be considered perspective dietary compounds purposed for prevention and treatment of chronic lead poisoning. PMID- 16717036 TI - Developmental toxicity study of pure trans-capsaicin in rats and rabbits. AB - Human environmental and dietary exposure to trans-capsaicin--the pungent ingredient in chili peppers--is ubiquitous. Moreover, based on the highly selective agonism of trans-capsaicin for TRPV1 receptors, drug products containing high concentrations of trans-capsaicin are under development as analgesics. For instance, a high-concentration (8% w/w) pure trans-capsaicin dermal patch (designated NGX-4010) is in advanced clinical evaluation for the management of neuropathic pain of peripheral origin. Our objective was to investigate effects of trans-capsaicin on embryo/fetal development, consequent to maternal exposure, from implantation to closure of the hard palate. trans Capsaicin was delivered systemically by means of either a patch [NGX-4010 (25, 37.5, or 50 cm(2))] to pregnant Sprague-Dawley rats on days of presumed gestation (DGs) 7 through 17, or via a 10% w/v capsaicin liquid formulation (CLF), at dosages of 3, 6.5 or 13 mul/cm(2) applied to a 200-cm(2) area on the back on DGs 7 though 19 to timed-mated New Zealand white rabbits. In rats, the maternal no observable-effect level (NOEL) was less than 25 cm(2) but no cesarean-sectioning or litter parameters were affected by application of NGX-4010 at patch sizes as high as 50 cm(2). The only test article-related observations were delays in skeletal ossification, evident as significant reductions in the average number of metatarsals and ossified hindlimb and forelimb phalanges that occurred in the 50 cm(2) NGX-4010 dose group. Although the values for ossified metatarsals were outside the historical control range, ossified hindlimb and forelimb phalanges were within historical control ranges. No other gross external, soft tissue, or skeletal fetal alterations (malformations or variations) were caused by application of the NGX-4010. In rabbits, the maternal NOEL was less than 3 mul/cm(2) CLF (or 0.3 mg/cm(2)trans-capsaicin) per 200 cm(2), but no cesarean sectioning or litter parameters were affected. No fetal alterations (malformations or variations) were caused by dosages of CLF as high as 13 mul/cm(2) (or 1.3 mg/cm(2)trans-capsaicin). Taken together, these data suggest that tran s-capsaicin should not be considered a developmental toxicant. PMID- 16717039 TI - The Balkan Piedmont: male suicide rates pre-war, wartime, and post-war in Serbia and Montenegro. AB - The epidemiology of suicide in Serbia and Montenegro from 1989 to 2003, a period of civil war, is presented. Following the break-up of former Yugoslavia, Serbia and Montenegro underwent a period of war from 1991-1994 and another in 1999. During the war years, the number of suicides increased, reaching its peak in 1993. Male suicides outnumbered female suicides by a ratio of 2:1. Male suicides decreased slightly after the war of 1991-1994 only to rise in 1997 and continue at this higher level throughout the nineties. In Serbia alone, male suicide reached its peak in 2002 (nearly 29/100,000). The methods of suicide changed significantly, with the use of firearms doubling during and after the war years. Speculations are offered about the findings, many consistent with Durkheim's classical hypothesis concerning suicide and unpopular wars. PMID- 16717037 TI - Oncogenicity evaluations of chemopreventive soy components in p53((+/-)) (p53 knockout) mice. AB - Epidemiologic data suggest that soy consumption may protect against cancer induction in several tissues in humans. Although the soy components responsible for this activity remain unidentified, isoflavones (e.g., genistein) and protease inhibitors (e.g., Bowman-Birk inhibitor complex [BBIC]) demonstrate chemopreventive activity in several animal cancer models. As part of their preclinical development for cancer prevention, PTI G-2535 (a soy isoflavone mixture containing 45% genistein, 23% daidzein, and 4% glycitein) and BBIC were evaluated for oncogenicity in p53((+/-)) mice. In separate studies, groups of 25 p53((+/-)) mice/sex received daily gavage exposure to PTI G-2535 (0, 250, 1000, or 2500 mg/kg/day) or BBIC (0, 500, 1000, or 2000 mg/kg/day) for 6 months. The high doses of both PTI G-2535 and BBIC were limited by viscosity. p-Cresidine (400 mg/kg/day) served as a positive-control article in both studies. PTI G-2535 induced no gross toxicity in any animal, but did induce a dose-related suppression of body weight gain in male mice. Modest hematologic alterations and increased liver and spleen weights were seen in both sexes exposed to the isoflavone mixture. BBIC had no significant effect on body weight, food consumption, clinical pathology, or organ weights in either sex. Histopathologic evaluations demonstrated no increases in the incidence of either benign or malignant tumors in any group of p53((+/-)) mice exposed to PTI G-2535 or to BBIC. By contrast, the positive-control article, p-cresidine, induced urinary bladder cancers in both studies. Neither PTI G-2535 nor BBIC demonstrates any evidence of oncogenicity in the p53((+/-)) mouse model. PMID- 16717040 TI - Country of birth and suicide: a follow-up study of a national cohort in Sweden. AB - The aim of this study was to examine whether the hypothesized association between country of birth and suicide rates remains after adjustment for age, marital status, socioeconomic status, and hospitalization due to psychiatric disorders or substance abuse. A Swedish cohort of 4.4 million individuals aged 25-64 years was followed from January 1, 1994, to December 31, 1999 for suicide. Suicide rates varied according to country of birth. Among men the highest risk of suicide was found among men from Finland. Among women the highest risk of suicide was found among women from Finland, Poland, and Eastern Europe. Key factors to prevent suicide include early detection and treatment of psychiatric disorders and/or substance abuse, especially among certain population groups. PMID- 16717041 TI - Rating suicidality in schizophrenia: items on global scales (HoNOS and CPRS) correlate with a validated suicidality rating scale (InterSePT). AB - The use of general scales such as the Health of the Nation Outcome Scale (HoNOS) and the Comprehensive Psychopathological Rating Scale (CPRS) are widespread. Both scales contain non-validated items on suicidality. This study aimed to establish whether an association between these suicidality items and the validated scale, InterSePT exists. Forty patients with a diagnosis of schizophrenia or schizo affective disorder were assessed using the three scales. A highly significant association between the two items and the validated scale was found. The suicidality items from HoNOS and CPRS appear to be strong indicators of suicidality in this patient population. The findings allow for use of the suicidality items in clinical assessments and re-investigation of suicidality of other studies where these general rating scales have been used. PMID- 16717042 TI - The accuracy of prevalence estimations for suicide attempts. how reliably do adolescents and young adults report their suicide attempts? AB - This study explores the accuracy of prevalence estimations for suicide attempts. Data came from the Early Developmental Stages of Psychopathology (EDSP) Study, a prospective community study (mean follow-up period was 42 months) of 3,021 respondents aged 14 to 24 years at the outset of the study. Suicide attempters are at least 1.6 times more likely to drop out than subjects with no suicide attempts and suicidal ideas. A total of 8% of all suicide attempters answered in the negative the depression-related gate questions of all surveys. One-third of all baseline suicide attempters did not report their suicide attempt again at the four years later assessment. In particular, 80% of all nonreporters were female, and almost 60% were aged 14-17 at baseline. PMID- 16717043 TI - Adolescent perceptions of a suicide prevention group on an inpatient unit. AB - The purpose of this study was to present a psychoeducational suicide prevention group for psychiatrically hospitalized suicidal adolescents, and explore participant perceptions of the strengths and weaknesses of this group therapy experience using archival data. Over the course of approximately three years, 250 adolescents admitted to a psychiatric inpatient unit attended the suicide prevention group. Almost all adolescents indicated that they learned something helpful in the group that would keep them from attempting suicide in the future. The creation of a "Reasons To Live List" was reported as most helpful by adolescents while the creation of a "Safety List" was deemed least helpful. However, some differences did emerge as a function of history of prior suicidal behavior. Recommendations for future treatment research with suicidal adolescent psychiatric inpatients are offered. PMID- 16717044 TI - A study of multi-media suicide education in Nunavut. AB - A CD-ROM on suicide prevention was evaluated in an Inuit hamlet to examine if knowledge is acquired after viewing this multi-media format of education. Twenty four respondents showed knowledge increased after viewing. Forty six percent had never used a computer yet 95% felt the technology could train counselors. Such technology may improve knowledge and skills in this cultural setting and perhaps may be useful in other cross-cultural environments. PMID- 16717045 TI - Suicidality in bipolar disorders--psychoanalytic contribution. AB - This paper gives an overview of psychoanalytic contributions to the understanding of suicidal behavior in bipolar patients. Although little specific literature is available, many authors have contributed to the understanding of these patients' psychodynamics and suicidality in various papers. Different points of emphasis are described, among these are defensive strategies, narcissistic personality structure, and dealing with intense feelings such as object loss. Using detailed case descriptions, the inner world of bipolar patients as it relates to their suicidality, along with the appropriate psychoanalytically oriented approach to treatment, are highlighted. PMID- 16717046 TI - Understanding suicide through studies of diaries: the case of Cesare Pavese. AB - The diary left by Cesare Pavese covering the 15 years prior to his suicide is examined. The noteworthy pattern is that mention of failure with women was immediately followed in his diary by denigration of his literary work. Other features of his life, such as the loss of his father when he was six, are also discussed. PMID- 16717047 TI - The impact of lithium long-term medication on suicidal behavior and mortality of bipolar patients. PMID- 16717048 TI - Two wrongs don't make a right. PMID- 16717049 TI - Morbidity in expatriates--a prospective cohort study. AB - BACKGROUND: Expatriates comprise an important, but rarely studied subset of international travellers. This study was performed to assess the incidence of health events in an expatriate group and to evaluate factors affecting this incidence. METHODS: A cohort of 2020 Foreign and Commonwealth Office (FCO) staff and partners living abroad were followed-up over 1 year. The main outcome measure was incidence of illness or injury serious enough to require consultation with a doctor. Data collection was by means of a self-administered questionnaire. Poisson regression was used to estimate the rates of health events and to test for association between health events and a number of independent variables. RESULTS: The incidence of health events was 21%. Trauma (incidence 5%), musculoskeletal disorders (incidence 4%) and infectious disease (incidence 3%) were the principal causes of morbidity. The incidence of psychological disorders was low (1%). Of significance, employees were at increased risk of morbidity when compared to partners, with a higher incidence of health events [incidence rate ratio (IRR) 1.4, 95% CI 1.1-1.9] and psychological disorders (IRR 5.9, 95% CI 1.0 34.1). Moreover, unaccompanied employees were at increased risk of health events (IRR 1.3, 95% CI 1.0-1.7), and of traumatic injury (IRR 2.3, 95% CI 1.3-4.3) when compared to accompanied employees. CONCLUSION: While the morbidity in FCO personnel is low in comparison to other expatriate groups, the higher risk of morbidity in employees and unaccompanied individuals merits further research, particularly to ascertain whether work demands, isolation or risk-taking behaviour are contributory factors. PMID- 16717050 TI - Vibration exposure and myocardial infarction incidence: the VHEEP case-control study. AB - OBJECTIVES: The main objectives of this study were to assess the risk of contracting first episode of myocardial infarction (MI) subsequent to vibration exposure and to assess a possible exposure-response relationship. METHODS: The Vasternorrland heart epidemiology programme (VHEEP, a part of the Stockholm heart epidemiology programme study) was the source of the data. VHEEP is a population based case-control study of risk factors for acute MI. Exposure information was collected by questionnaire and vibration exposure was assessed in 218 cases and 257 controls. Relative risks were estimated using odds ratios (ORs) from binary logistic regression. RESULTS: The results show that the OR of acute MI when exposed to vibration was 1.6 (95% CI: 1.1-2.4). It was not possible, however, to determine whether an exposure-response relationship was present. CONCLUSIONS: Working entailing vibrating machines is associated with an increased risk for acute MI. PMID- 16717051 TI - Hand-arm vibration syndrome with concomitant arterial thrombosis in the hands. AB - BACKGROUND: Hand-arm vibration syndrome (HAVS) refers to the vascular, neurological and musculoskeletal effects that may occur in workers with prolonged exposure to vibrating tools. Hypothenar hammer syndrome (HHS) is a lesion of the ulnar artery at the level of the hamate bone secondary to single or repeated episodes of trauma to the hypothenar eminence. The literature suggests that digital arterial thrombosis and HHS may be associated with the use of vibrating tools. AIM: This study will familiarize investigators with the range of vascular abnormalities seen in workers using vibrating tools, and highlight the importance of screening for arterial thrombosis in the hand when assessing hand-arm vibration-exposed patients. METHODS: In the patients referred to our clinic for HAVS assessment, three were identified during the period 2001 to 2004 who had vascular occlusions in the hands in addition to HAVS. In addition to standardized HAVS vascular investigations, all three patients had arteriograms based on a significantly positive Allen's test. RESULTS: All three cases had documented HAVS based on vascular testing. Arteriograms revealed a spectrum of severity of arterial thromboses from severe HHS, to occlusion limited to the digital arteries. CONCLUSION: Our study reports three cases of HAVS with concomitant HHS and/or digital artery thrombosis. These findings support previous reports of an association between HAVS and vascular thrombosis in the hands. Screening for arterial occlusive problems in the hands should be included in the HAVS work up. PMID- 16717052 TI - Comment on: suboptimal CD4 gains in HIV-infected patients receiving didanosine plus tenofovir. PMID- 16717053 TI - Pharmacokinetic interaction between methotrexate and piperacillin/tazobactam resulting in prolonged toxic concentrations of methotrexate. PMID- 16717054 TI - Dissimilar distribution of Trypanosoma cruzi clones in humans after chemotherapy with allopurinol and itraconazole. AB - OBJECTIVES: The aim of this work was to study the distribution of Trypanosoma cruzi clones after treatment failure with itraconazole or allopurinol in infected humans. METHODS: Blood samples from treated and untreated individuals were used to detect T. cruzi by PCR assays and were confirmed by hybridization tests using total kinetoplast DNA as a universal probe. Also, xenodiagnosis (XD) tests were performed with Triatoma infestans fed from the same group of patients. We performed Southern-blot analyses of PCR products from blood or XD samples using a panel of four genotype-specific probes: corresponding to T. cruzi clones TcI, TcIIb, TcIId and TcIIe. The membranes were hybridized with radiolabelled probes and exposed in a Personal Molecular Imager. RESULTS: When comparing the presence of T. cruzi clones in the allopurinol-treated group with the non-treated group significant differences were only observed for XD samples. Clone TcI was present in 9/13 (69.2%) of the XD samples of the treated group, but only in 8/27 (29.6%) in the non-treated group (P = 0.0178). When the itraconazole-treated group and the control group were compared, significant differences were found in both the blood and XD samples. In blood, the clone TcIIb was detected in 6/17 (35.5%) of the treated group and in 18/27 (66.7%) of the non-treated group (P = 0.0207). When XD samples were analysed, the clone TcI was observed in 14/17 (82.3%) of the itraconazole-treated group but only in 8/27 (29.6%) of the control group (P = 0.0006), which suggests resistance of this clone to itraconazole. CONCLUSIONS: We detected a dissimilar distribution of T. cruzi clones in treated and untreated groups of patients. The presence of TcI increased in patients treated with allopurinol and itraconazole, whereas the presence of TcIIb decreased in itraconazole-treated patients. The type of T. cruzi clone that prevails suggests that TcI is resistant to both drugs and that TcIIb is susceptible to itraconazole. PMID- 16717055 TI - In vitro susceptibilities of aerobic and facultatively anaerobic Gram-negative bacilli isolated from patients with intra-abdominal infections worldwide: 2004 results from SMART (Study for Monitoring Antimicrobial Resistance Trends). AB - OBJECTIVES: SMART (Study for Monitoring Antimicrobial Resistance Trends) is an ongoing study to monitor worldwide antimicrobial resistance trends among aerobic and facultatively anaerobic Gram-negative bacilli (GNB) isolated from intra abdominal infections. This 2004 report summarizes the most recently completed annual data from SMART. METHODS: During 2004, 81 medical centres from 28 countries in five global regions collected intra-abdominal GNB for antimicrobial susceptibility testing using broth microdilution according to the Clinical and Laboratory Standards Institute guidelines. RESULTS: A total of 6156 unique aerobic and facultatively anaerobic GNB were isolated from intra-abdominal infections. Enterobacteriaceae composed 86% of the total isolates. Among the 12 antimicrobial agents tested, the carbapenems and amikacin were the most consistently active against the Enterobacteriaceae. Escherichia coli was the most commonly isolated species (48%), and the susceptibility rate to the quinolones was lowest in Asia/Pacific and Latin America. Extended-spectrum beta-lactamases (ESBLs) were detected phenotypically in 10% of E. coli, 17% of Klebsiella spp. and 22% of Enterobacter spp. worldwide, representing a slight increase over the two previous years. ESBL producers typically had a more antibiotic-resistant profile than non-ESBL producers but were usually susceptible to the carbapenems. CONCLUSIONS: Antimicrobial resistance among GNB isolated from intra-abdominal infections continued to be a problem worldwide in 2004, with the highest rates of resistance overall in the Asia/Pacific region. The carbapenems and amikacin were the most consistently active agents in vitro against Enterobacteriaceae isolated from intra-abdominal infections worldwide. PMID- 16717056 TI - Contextualizing salutogenesis and Antonovsky in public health development. AB - More than 20 years have passed since the American-Israeli medical sociologist Aaron Antonovsky introduced his salutogenic theory 'sense of coherence' as a global orientation to view the world, claiming that the way people view their life has a positive influence on their health. Sense of coherence explains why people in stressful situations stay well and even are able to improve their health. The origin of salutogenesis derives from the interviews of Israeli women with experiences from the concentration camps of the Second World War who in spite of this stayed healthy. Sixty years after the Holocaust this paper aim to shed light on the salutogenic theory in the context of public health and health promotion. In addition, other approaches with salutogenic elements for the explanation of health are considered. A potential direction for public health of the early 21st century is proposed. The historical paradox is to honour the victims of the Holocaust and see the birth of post-modern public health and the salutogenic framework through the experience of its survivors in the ashes of Modernity. PMID- 16717057 TI - Fusion between CIC and DUX4 up-regulates PEA3 family genes in Ewing-like sarcomas with t(4;19)(q35;q13) translocation. AB - Ewing's family tumors (EFTs) are highly malignant tumors arising from bone and soft tissues that exhibit EWS-FLI1 or variant EWS-ETS gene fusions in more than 85% of the cases. Here we show that CIC, a human homolog of Drosophila capicua which encodes a high mobility group box transcription factor, is fused to a double homeodomain gene DUX4 as a result of a recurrent chromosomal translocation t(4;19)(q35;q13). This translocation was seen in two cases of soft tissue sarcoma diagnosed as Ewing-like sarcoma. CIC-DUX4 exhibits a transforming potential for NIH 3T3 fibroblasts, and as a consequence of fusion with a C-terminal fragment of DUX4, CIC acquires an enhanced transcriptional activity, suggesting that expression of its downstream targets might be deregulated. Gene expression analysis identified the ETS family genes, ERM/ETV5 and ETV1, as potential targets for the gene product of CIC-DUX4. Indeed, CIC-DUX4 directly binds the ERM promoter by recognizing a novel target sequence and significantly up-regulates its expression. This study clarifies the function of CIC and its role in tumorigenesis, as well as the importance of the PEA3 subclass of ETS family proteins in the development of EFTs arising through mechanisms different from those involving EWS-ETS chimeras. Moreover, the study identifies the role of DUX4 that is closely linked to facioscapulohumeral muscular dystrophy in transcriptional regulation. PMID- 16717058 TI - TRPV6 exhibits unusual patterns of polymorphism and divergence in worldwide populations. AB - A striking footprint of positive selection was recently identified on chromosome 7q34-35 that spans at least 115 kb and encompasses four known genes (KEL, TRPV5, TRPV6, EPHB6). The signature of selection was observed in only one of the two populations analyzed suggesting the action of geographically restricted selective pressures. However, as only two populations were analyzed, it remains unknown whether the signature of selection extends to additional populations. To address this issue and begin to dissect the evolutionary history of this region in more detail, we performed an in-depth population genetic analysis on TRPV6, which is a calcium-permeable ion channel thought to mediate the rate-limiting step of dietary calcium absorption. We demonstrate that the rate of TRPV6 protein evolution is significantly accelerated in the human lineage, but only for a haplotype defined by three non-synonymous SNPs (C157R, M378V and M681T) that are nearly fixed for the derived alleles in non-African populations. Interestingly, we found that these three non-synonymous SNPs have high posterior probabilities for being targets of positive selection and are therefore strong candidates for mediating the population-specific signatures of selection in this region. In addition, we resequenced the exons corresponding to the C157R, M378V and M681T polymorphisms in 90 geographically diverse individuals and characterized their global allele frequency distribution by genotyping them in 1064 individuals from 52 populations. These data strongly suggest that the TRPV6 haplotype defined by the derived alleles at C157R, M378V and M681T conferred a selective advantage that varied spatially, and perhaps temporally, during human history. PMID- 16717059 TI - Failure of MBNL1-dependent post-natal splicing transitions in myotonic dystrophy. AB - In myotonic dystrophy (DM), expression of RNA containing expanded CUG or CCUG repeats leads to misregulated alternative splicing of pre-mRNA. The repeat bearing transcripts accumulate in nuclear foci, together with proteins in the muscleblind family, MBNL1 and MBNL2. In transgenic mice that express expanded CUG repeats, we show that the splicing defect selectively targets a group of exons that share a common temporal pattern of developmental regulation. These exons undergo a synchronized splicing switch between post-natal day 2 and 20 in wild type mice. During this post-natal interval, MBNL1 protein translocates from a predominantly cytoplasmic to nuclear distribution. In the absence of MBNL1, these physiological splicing transitions do not occur. The splicing defect induced by expanded CUG repeats in mature muscle fibers is closely reproduced by deficiency of MBNL1 but not by deficiency of MBNL2. A parallel situation exists in human DM type 1 and type 2. MBNL1 is depleted from the muscle nucleoplasm because of sequestration in nuclear foci, and the associated splicing defects are remarkably similar to those observed in MBNL1 knockout mice. These results indicate that MBNL1 participates in the post-natal remodeling of skeletal muscle by controlling a key set of developmentally regulated splicing switches. Sequestration of MBNL1, and failure to maintain these splicing transitions, has a pivotal role in the pathogenesis of muscle disease in DM. PMID- 16717060 TI - Reduced expression of top1beta gene induces programmed cell death and alters ascorbate metabolism in Daucus carota cultured cells. AB - Topoisomerase I (topo I) is a nuclear enzyme which plays a fundamental role in several pathways involving changes in DNA topology. The topo I-mediated reaction is accomplished by the transient covalent binding of the enzyme to DNA (topo I DNA complex). Stabilization of the topo I-DNA complex, leading to irreversible double-strand breaks, has been reported to occur in animal cells under oxidative stress conditions and during apoptosis. In order to study the existence of a putative link between the topo I-mediated DNA damage and ascorbate (ASC) metabolism, also involved in the responses against oxidative stress and in the apoptotic process in plants, Daucus carota cells showing reduced expression of the top1beta gene encoding the topo Ibeta isoform were produced, using an antisense RNA strategy. Two independent transgenic lines (AT1-beta/22 and beta/36), characterized by a slow growth phenotype, resistance to camptothecin, a specific inhibitor of topo I, but sensitivity to etoposide, an inhibitor of topo II, were investigated in this study. In the absence of external stimuli, AT1 beta/22 and beta/36 cells underwent programmed cell death (PCD) in a precocious phase of the growth curve. ASC metabolism showed remarkable differences in AT1 beta/22 and beta/36 cells, compared with control, and the observed alterations were similar to those occurring in tobacco Yellow Bright-2 cells induced to enter PCD by exogenous stimuli. However, differently from other studied examples of PCD, overproduction of reactive oxygen species was not detected in AT1-beta/22 and beta/36 cells. The relevance of these findings in relation to the signalling pathways leading to PCD is discussed. PMID- 16717062 TI - Re: Touch screen computer systems in the rheumatology clinic offer a reliable and user-friendly means of collecting quality-of-life and outcome data from patients with rheumatoid arthritis. PMID- 16717063 TI - Polymorphisms of the FCRL3 gene in a Spanish population of systemic lupus erythematosus patients. PMID- 16717064 TI - Patients' own ability to assess activity of their rheumatoid arthritis. PMID- 16717065 TI - A patient-centred approach to drug regulation. PMID- 16717066 TI - Severe infections following leflunomide therapy for rheumatoid arthritis. PMID- 16717067 TI - Metabolic syndrome and risk for heart failure in middle-aged men. AB - OBJECTIVE: To explore metabolic syndrome as a possible risk factor for development of heart failure (HF). DESIGN: Community-based cohort study. SETTING: Uppsala, Sweden. PARTICIPANTS: 2314 50-year-old men free from HF, myocardial infarction and valvular disease at baseline were enrolled between 1970 and 1974 and were followed up until the age of 70. A modified National Cholesterol Education Program definition of metabolic syndrome was used with body mass index in the place of waist circumference. MAIN OUTCOME MEASURE: First hospitalisation for HF. RESULTS: In multivariable Cox proportional hazards models adjusted for established risk factors for HF (hypertension, diabetes, ECG left ventricular hypertrophy, smoking and body mass index), the presence at baseline of metabolic syndrome (hazard ratio 1.66, 95% confidence interval (CI) 1.02 to 2.70) was a predictor of subsequent HF. This relation was even stronger after adjustment for the presence of an acute myocardial infarction during follow up in addition to the other established risk factors for HF (hazard ratio 1.80, 95% CI 1.11 to 2.91). CONCLUSION: Metabolic syndrome was a significant predictor of HF, independent of established risk factors for HF including an interim myocardial infarction, during two decades of follow up in a community-based sample of middle aged men. This implies that metabolic syndrome provides important risk information beyond that of established risk factors for HF. PMID- 16717068 TI - American, British and European recommendations for statins in the primary prevention of cardiovascular disease applied to British men studied prospectively. AB - OBJECTIVE: To compare national and international recommendations for statin treatment in the primary prevention of cardiovascular disease (CVD) in middle aged men. DESIGN: Application of the current American, British and European recommendations to results of a prospective study. PARTICIPANTS: Men aged 49-65 years (n = 1653) who participated in the Caerphilly Prospective Study. MAIN OUTCOME MEASURES: Proportion of patients who would receive statin treatment, the number needed to treat (NNT) to prevent one first CVD event (myocardial infarction or stroke) over 10 years and the potential number of events prevented over 10 years in the whole population (population impact) by the use of statins in accordance with each set of guidelines, assuming a reduction of risk in the range 10-50% from the observed events and baseline risk factors. RESULTS: 212 events were noted. For an anticipated reduction in first CVD events of 30% with statin treatment, the NNT was 26.0, if the whole population was treated. The lowest NNT was 12.1 for the National Service Framework, achieved when only 14% of the men received a statin. This prevented the lowest number of events (19.2/212), however, and had the smallest population impact on CVD incidence (-9.1%). The American and earlier Joint British Societies guidelines, although giving NNTs of around 21, prevented more events and had a greater population impact of -21.6% to -23.3%. They did, however, target about 60% of the male population. The British Hypertension Society guidelines and new Joint British Societies recommendations achieved the greatest population impact of -27% while maintaining the NNT at 22.2. They did, however, target three quarters of this population. CONCLUSION: Even effective preventive treatment will have little impact in preventing disease if patients at typical risk are not treated. Whether cholesterol lowering on such a scale should be attempted with drugs raises philosophical, psychological and economic considerations, particularly in view of the high likelihood of individual benefit from statin treatment. More effective nutritional policies to reduce serum cholesterol on a population level and reduce the requirement for statins in primary prevention should also be considered. PMID- 16717069 TI - Non-steroidal anti-inflammatory drugs and risk of first hospital admission for heart failure in the general population. AB - OBJECTIVES: To estimate the risk of a first hospital admission for heart failure (HF) associated with the use of non-steroidal anti-inflammatory drugs (NSAIDs). METHODS: Cohort study with a nested case-control analysis based on the UK General Practice Research Database. Overall, 1396 cases of first hospital admission for non-fatal HF were identified (January 1997 to December 2000) and compared with a random sample of 5000 controls. RESULTS: The incidence rate was 2.7/1000 person years. Prior clinical diagnosis of HF was the main independent risk factor triggering a first HF hospitalisation (relative risk 7.3, 95% confidence interval (CI) 6.1 to 8.8). The risk of a first hospital admission for HF associated with current use of NSAIDs was 1.3 (95% CI 1.1 to 1.6) after controlling for major confounding factors. No effects of dose and duration were found. The relative risk in current users of NSAIDs with prior HF was 8.6 (95% CI 5.3 to 13.8) compared with patients who did not use NSAIDs and without prior clinical diagnosis of HF. CONCLUSION: Use of NSAIDs was associated with a small increase in risk of a first hospitalisation for HF. In patients with prior clinical diagnosis of HF, the use of NSAIDs may lead to worsening of pre-existing HF that triggers their hospital admission. This increased risk, although small, may result in considerable public health impact, particularly among the elderly. PMID- 16717070 TI - NETGEN: generating phylogenetic networks with diploid hybrids. AB - SUMMARY: NetGen is an event-driven simulator that creates phylogenetic networks by extending the birth-death model to include diploid hybridizations. DNA sequences are evolved in conjunction with the topology, enabling hybridization decisions to be based on contemporary evolutionary distances. NetGen supports variable rate lineages, root sequence specification, outgroup generation and many other options. This note describes the NetGen application and proposes an extension of the Newick format to accommodate phylogenetic networks. AVAILABILITY: NetGen is written in C and is available in source form at http://www.phylo.unm.edu/~morin/. PMID- 16717071 TI - Prognostic value of circulating pregnancy-associated plasma protein levels in patients with chronic stable angina. AB - AIMS: Unstable coronary atherosclerotic plaque can be present in patients with chronic stable coronary artery disease (CAD). Our objective was to assess whether measurement of plasma pregnancy-associated plasma protein (PAPP-A) level, a reflection of plaque instability, in patients with chronic stable CAD had an independent prognostic value on the subsequent incidence of death, acute coronary syndrome (ACS), and revascularization. METHODS AND RESULTS: Patients referred for coronary angiography were recruited. A cohort of 103 patients with stable symptoms for at least 6 weeks and with a coronary angiogram showing at least a 50% luminal diameter narrowing formed our study population. Median follow-up was 4.9 years. Mean age was 65+/-10 years. In a multivariable model that included CAD traditional risk factors, ejection fraction, extent of coronary atherosclerosis, prior history of myocardial infarction, prior revascularization, discharge medications, and C-reactive protein, the plasma PAPP-A was found to be significantly associated with the endpoint of future death [adjusted hazard ratio (HR) 5.29; 95% CI 1.27-22.0; P=0.023] and with the endpoint of future death and ACS (adjusted HR 3.56; 95% CI 1.27-10.0; P=0.015), but not with the endpoint of future death and revascularization. CONCLUSION: Measurement of plasma PAPP-A level in patients with chronic stable CAD has an independent prognostic value on the occurrence of death and ACS. PMID- 16717073 TI - What to 'eat and chew' in acute myocardial infarction? PMID- 16717072 TI - Quality of life after coronary revascularization in the elderly. AB - AIMS: To describe health status outcomes at 4 years for a cohort of elderly patients with cardiac disease. METHODS AND RESULTS: Using the Alberta Provincial Project for Outcomes Assessment in Coronary Heart Disease, an outcomes initiative capturing all patients undergoing cardiac catheterization in Alberta, Canada, health status was measured using the Seattle Angina Questionnaire (SAQ) and crude and risk-adjusted outcomes were determined and compared for patients treated with percutaneous coronary intervention or coronary artery bypass surgery (CABG) vs. medical therapy. Response rates among surviving, consenting patients were 64.8% for patients <70 years (n=7883), 77.3% for patients aged 70-79 years (n=2940), and 77.7% for patients >or=80 years of age (n=439). For patients aged <70 years, and those aged 70-79 years, for all dimensions of the SAQ, scores were significantly better for patients treated with revascularization procedures than with medical therapy. For patients over the age of 80 years, scores for patients treated with CABG in particular were significantly better, with the exception of exertional capacity. At 3 years, all scores remained stable or improved, and continued to favour revascularization. CONCLUSION: Elderly patients undergoing revascularization have better health status at 4 years than do those in the same age group who do not undergo revascularization. These findings suggest that age should not deter against revascularization given the combined survival and quality-of-life benefits. PMID- 16717074 TI - Relation between baseline plaque burden and subsequent remodelling of atherosclerotic left main coronary arteries: a serial intravascular ultrasound study with long-term (> or =12 months) follow-up. AB - AIMS: Glagov's histopathological observation and non-serial intravascular ultrasound studies (IVUS) concluded that compensatory coronary remodelling diminishes as 40% atherosclerotic plaque burden is reached. We tested this hypothesis with serial IVUS. METHODS AND RESULTS: Serial IVUS examinations of 46 atherosclerotic non-stenotic left main stems (18+/-8 months apart) were analysed to assess the relation between baseline plaque burden (=plaque+media area/vessel area) vs. serial remodelling (=vessel area at baseline - at follow-up). There were 25 plaques with baseline plaque burden <40% (30.1+/-6.6%, group A) and 21 plaques with baseline plaque burden > or =40% (46.1+/-5.8%, group B). There was no relation between baseline plaque burden vs. subsequent changes in vessel area overall (r=0.07, P=0.7), for group A (r=0.03, P=0.6), and group B (r=0.01, P=0.8). The frequency of positive serial remodelling (vessel area increase) vs. negative or intermediate serial remodelling (no change or decrease) were similar in group A [17 (68%) vs. 8 (32%)] and group B lesions [18 (86%) vs. 3 (14%)] (P=0.2). CONCLUSION: IVUS demonstrates that serial coronary remodelling is not related to baseline plaque burden. Lesions with baseline plaque burden <40% may subsequently show a lack of compensation or frank arterial shrinkage, whereas lesions with baseline plaque burden >40% may continue to develop compensatory enlargement. PMID- 16717075 TI - Detection of myocardial infarction by delayed contrast-enhanced multislice computed tomography. PMID- 16717076 TI - Arrhythmogenic right ventricular dysplasia/cardiomyopathy assessed with 64-slice computed tomography. PMID- 16717077 TI - Molecular analysis of pericardial fluid: a 7-year experience. AB - AIMS: Aetiological investigations of pericardial effusion are often negative. We evaluate the interest of systematic analysis of fluid in order to diminish the number of pericarditis classified as idiopathic. METHODS AND RESULTS: We performed a systematic analysis of pericardial fluid and biopsy specimens, using cultures and molecular analyses for the identification of bacteriological, fungal, and viral agents, as well as histopathological examination of 106 pericardial fluid samples. The aetiological diagnosis was determined clinically and by non-invasive procedures in 40 and nine patients, respectively. In the remaining 57 patients, 14 neoplasias and 17 infections were diagnosed. Molecular procedures identified seven viral (Enterovirus, Herpes simplex virus, and Epstein Barr virus in four, two, and one of the cases, respectively), one fungal (Cryptococcus neoformans), and nine bacterial infections. Four of these bacteria were not diagnosed by culture because of prior antibiotics treatment (Mycobacterium tuberculosis in two cases, Streptococcus pneumoniae in one case, and Actinomyces neuii in one case). The aetiology remained undetermined in 26 patients. CONCLUSION: The systematic use of molecular techniques permitted a significant increase in aetiological diagnoses of punctured pericardial effusions when compared with cultures (39.5 vs. 13.9%, respectively; P<0.01). It is particularly beneficial for patients with a previous antibiotic regimen or suspicion of tuberculosis. PMID- 16717078 TI - The relative myocardial blood volume differentiates between hypertensive heart disease and athlete's heart in humans. AB - AIMS: The adaptation of the myocardial microcirculation in humans to pathologic and physiologic stress has not been examined in vivo so far. We sought to test whether the relative blood volume (rBV) measured by myocardial contrast echocardiography (MCE) can differentiate between left ventricular (LV) hypertrophy (LVH) in hypertensive heart disease and athlete's heart. METHODS AND RESULTS: Four groups were investigated: hypertensive patients with LVH (n = 15), semi-professional triathletes with LVH (n = 15), professional football players (n = 15), and sedentary control individuals without cardiovascular disease (n = 15). MCE was performed at rest and during adenosine-induced hyperaemia. The rBV (mL mL(-1)), its exchange frequency (beta, min(-1)), and myocardial blood flow (mL min(-1) g(-1)) were derived from steady state and refill sequences of ultrasound contrast agent. Hypertensive patients had lower rBV (0.093 +/- 0.013 mL mL(-1)) than triathletes (0.141 +/- 0.012 mL mL(-1), P < 0.001), football players (0.129 +/- 0.014 mL mL(-1), P < 0.001), and sedentary individuals (0.126 +/- 0.018 mL mL(-1), P < 0.001). Conversely, the exchange frequency (beta) was significantly higher in hypertensive patients (11.3 +/- 3.8 min(-1)) than in triathletes (7.4 +/- 1.8 min(-1)), football players (7.7 +/- 2.3 min(-1)), and sedentary individuals (9.0+/-2.5 min(-1)). An rBV below 0.114 mL mL(-1) distinguished hypertensive patients and triathletes with a sensitivity of 93% and a specificity of 100%. CONCLUSION: Pathologic and physiologic LVH were differentiated non invasively and accurately by rBV, a measure of vascularisation assessed by MCE. PMID- 16717079 TI - Cardiac catheterization and long-term chromosomal damage in children with congenital heart disease. AB - AIMS: Medical radiological exposure is associated with an additional risk of cancer. Children with repaired congenital heart disease (CHD) are theoretically at a relatively greater cancer risk as the radiological exposure can be intensive in these patients. Chromosomal aberrations test (CA) and micronucleus assay (MN) in peripheral blood lymphocytes are biomarkers of chromosomal damage and intermediate endpoints in carcinogenesis. METHODS AND RESULTS: The frequency of CA and MN was assessed in three groups of patients: Group I, 32 exposed patients (17 males, age=15.5+/-8.3 years) who underwent cardiac procedures employing ionizing radiation (mostly cardiac catheterization) for CHD between 1965 and 2000; Group II, 32 healthy age- and sex-matched subjects (17 males, age=14.1+/ 12.3 years), and Group III, 10 newborn non-exposed patients (7 males) with CHD. Exposed patients of Group I had a mean value of 2.9+/-1.4 cardiac catheterization (range 1-5) procedures per person. The mean frequency of CA was higher in the exposed patients (Group I=2.8+/-1.9% vs. Group II=0.7+/-0.7%; vs. Group III=0.8+/ 0.8%; P<0.0001). Similarly, the mean values of MN were higher in the exposed patients (Group I =12.3+/-5.1 per thousand vs. Group II=6.0+/-3.8 per thousand; vs. Group III=4.4+/-1.4 per thousand; P<0.0001). CONCLUSION: Cardiac ionizing procedures are associated with a long-lasting mark in the chromosomal damage of exposed children with CHD. PMID- 16717081 TI - The impact of new onset anaemia on morbidity and mortality in chronic heart failure: results from COMET. AB - AIMS: Anaemia is a common comorbidity in chronic heart failure (CHF). The predictors of new onset anaemia (NOA) and its long-term prognostic value, particularly in patients treated with beta-blockers, are not known. METHODS AND RESULTS: In COMET, 3029 patients with CHF in NYHA II-IV and EF <35% were randomized to carvedilol or metoprolol tartrate and were followed for an average of 58 months. Plasma haemoglobin (Hb) concentrations were measured at a central laboratory at randomization, at four monthly intervals for the first year and annually thereafter. According to WHO criteria, anaemia was defined when Hb measured <13 g/dL for men and <12 g/dL for women. We considered anaemia to be severe when Hb <11.5 g/dL for men and <10.5 g/dL for women. The baseline mean Hb was 14.2 +/- 1.5 g/dL (n = 2996) and 15.9% of patients had anaemia (males, 16.0%; females, 15.2%). At baseline, severe anaemia was found in 3.3% of patients (males, 3.6%; females, 2.0%). During the study, all-cause mortality (RR 1.47) death or hospitalization (RR 1.28), and heart failure hospitalization (RR 1.43, all P < 0.0001) were higher in anaemic when compared with non-anaemic patients. In patients without anaemia at baseline, at the end of the study, the cumulative frequency of NOA was 28.1% in males and 27.0% in females. NOA increased over time from 14.2% at year 1 to 27.5% at year 5. Predictors of NOA were: higher age, diuretic dose, creatinine (all P < 0.0001), higher serum potassium, lower serum sodium, body mass index, and use of aldosterone antagonists, carvedilol, and digitalis (all P < 0.03). Treatment with carvedilol (vs. metoprolol tartrate) was associated with a 24% increased risk to develop NOA (P = 0.0047), but not severe anaemia (P = 0.18). Patients with a Hb decrease of >3 g/dL (RR 3.37, P < 0.0001) or of 2.0-3.0 g/dL (RR 1.47, P = 0.011) from baseline had an increased subsequent mortality when compared with patients having Hb increases of 0-1.0 g/dL. CONCLUSION: In stable ambulatory CHF patients, development of NOA is frequent and can be predicted by a set of clinical variables. Decreases in Hb over time relate to future increased morbidity and mortality. PMID- 16717080 TI - Tissue Doppler imaging predicts left ventricular dysfunction and mortality in a murine model of cardiac injury. AB - AIMS: Currently available non-invasive imaging methods frequently fail to detect alterations in left ventricular (LV) function despite histological evidence of injury. Tissue Doppler imaging (TDI) can detect subtle LV dysfunction. The aim of this study was to investigate whether TDI indices can predict LV systolic dysfunction and mortality following exposure to doxorubicin (DOX) in mice. METHODS AND RESULTS: TDI-derived peak endocardial systolic velocity (V(ENDO)) and strain rate (SR), as well as M-mode and two-dimensional indices of LV systolic function, were measured serially in mice after receiving DOX as a single dose (20 mg/kg). Haemodynamic measurements were obtained invasively before and at 1, 2, 4, and 5 days after the single DOX dose. Cardiac apoptosis was measured before and at 1 day after DOX. V(ENDO) and SR decreased after 1 and 2 days, respectively, whereas changes in fractional shortening (FS) and LV ejection fraction (LVEF) were not detected before 5 days. The reduction in both V(ENDO) and SR correlated with the decrease in dP/dt(MAX), and the change in V(ENDO) correlated with the early increase in cardiac cell apoptosis. In a subsequent experiment, DOX was administered at 4 mg/kg/week for 5 weeks, and LV function was followed serially for 16 weeks. In this chronic experiment, TDI indices decreased before FS and LVEF, correlated with late LV dysfunction, and predicted DOX-induced mortality. CONCLUSION: In a murine model of DOX-induced cardiac injury, TDI detects LV dysfunction prior to alterations in conventional echocardiographic indices and predicts mortality. This study suggests that TDI may be a reliable tool to detect early subtle changes in DOX-induced cardiac dysfunction. PMID- 16717082 TI - Accuracy and precision of test weighing to assess milk intake in newborn infants. AB - BACKGROUND: Test weighing is commonly used to estimate milk intake in newborn infants. OBJECTIVE: To assess the accuracy and precision of test weighing in clinical practice. METHODS: Infants fed by bottle, cup, or nasogastric tube were weighed before and immediately after feeding by a blinded investigator. Actual milk intake was determined by reading the millilitre scale of the milk container before and after feeding. The accuracy and precision of test weighing was assessed by examining the frequency distribution of the difference between weight change and actual milk intake. RESULTS: Ninety four infants completed the study. The mean difference between weight change and actual milk intake was 1.3 ml, indicating good accuracy. The precision of test weighing, however, was poor: 95% of differences between weight change and actual milk intake ranged from -12.4 to 15 ml. The maximum difference was 30 ml. Imprecision was not influenced by the presence of monitor or oxygen saturation wires, intravenous lines, or vomiting of the infant. CONCLUSIONS: Test weighing is an imprecise method for assessing milk intake in young infants. This is probably because infant weighing scales are not sensitive enough to pick up small changes in an infant's weight after feeding. Because of its unreliability, test weighing should not be used in clinical practice. PMID- 16717083 TI - Supply problems of unlicensed and off-label medicines after discharge. AB - A total of 709 unlicensed and off-label medicines were prescribed for 338 patients discharged from a paediatric hospital between 1 November 2003 and 31 January 2004. Thirty three per cent of patients had difficulty obtaining these medications in primary care which caused treatment disruption. The main problems were: (1) community pharmacies being unable to supply; and (2) GPs' refusal to prescribe. PMID- 16717084 TI - Managed Care Network for the assessment of cardiac problems in children in a district general hospital: a working model. AB - AIM: To assess a model for cardiology assessments in children with suspected heart disease by a general paediatrician with special expertise in paediatric cardiology (PsePC) in a district general hospital. METHODS: A new monthly "screening" clinic was established in May 2004 by the PsePC to reduce the burden of new referrals on outreach tertiary paediatric cardiology services. All patients were to have echocardiograms as part of their referral for cardiac assessment. Over a one year period (May 2004-April 2005), through audit, details of referrers, indications for referral, echocardiography assessments, and subsequent management were recorded. This was compared with the pattern of patients seen in the joint paediatric cardiology outreach clinics over a two year period (May 2003-April 2005). RESULTS: In the "screening" clinic, there were 75 appointments for 65 patients seen in 12 months. Fifty five of these patients had normal echocardiographic studies. Of the 47 referrals with heart murmurs in asymptomatic children, four had structurally abnormal hearts on echocardiographic assessment. Between May-October 2003 and November 2003-April 2004, the number of new patients with normal echocardiographic studies seen in the paediatric cardiology outreach clinic was 33/106 (31%) and 28/110 (25.4%) respectively. Following the introduction of the "screening" clinic, the number decreased to 21/99 (21%) during May-October 2004, and 10/102 (9.8%) during November 2004-April 2005. CONCLUSION: This model can work effectively in order to identify pathology requiring input of a paediatric cardiologist more appropriately. Paediatricians with specific training in paediatric cardiology are potentially well equipped to provide this basic screening service. PMID- 16717085 TI - Clinical and laboratory characteristics of non-E. coli urinary tract infections. AB - Comparison of the clinical and laboratory characteristics of infants and children with urinary tract infection caused by E. coli (n = 107) or other pathogens (n = 32) yielded a significantly higher association of non-E. coli disease with urinary tract anomalies, younger age, and previous antibiotic treatment. Underlying urinary tract anomalies were noted in 18 patients, of whom 14 (77%) were infected by non-E. coli pathogens. The most frequent anomaly was grade 3-4 vesicoureteral reflux (50%), followed by hydronephrosis (22.7%), ureteropelvic junction obstruction (9%), hypospadias (4.5%), pinpoint meatus (4.5%), and dysplastic kidney (4.5%). PMID- 16717086 TI - PP2A regulates BCL-2 phosphorylation and proteasome-mediated degradation at the endoplasmic reticulum. AB - Anti-apoptotic activity of BCL-2 is mediated by phosphorylation at the endoplasmic reticulum (ER), but how this phosphorylation is regulated and the mechanism(s) by which it regulates apoptosis are unknown. We purified macromolecular complexes containing BCL-2 from ER membranes and found that BCL-2 co-purified with the main two subunits of the serine/threonine phosphatase, PP2A. The association of endogenous PP2A and BCL-2 at the ER was verified by co immunoprecipitation and microcystin affinity purification. Knock down or pharmacological inhibition of PP2A caused degradation of phosphorylated BCL-2 and led to an overall reduction in BCL-2 levels. We found that this degradation was due to the action of the proteasome acting selectively at the ER. Conversely, overexpression of PP2A caused elevation in endogenous BCL-2. Most importantly, we found that PP2A knock down sensitized cells to several classes of death stimuli (including ER stress), but this effect was abolished in a genetic background featuring knock in of a non-phosphorylatable BCL-2 allele. These studies support the hypothesis that PP2A-mediated dephosphorylation of BCL-2 is required to protect BCL-2 from proteasome-dependent degradation, affecting resistance to ER stress. PMID- 16717087 TI - Utilization of an amphipathic leucine zipper sequence to design antibacterial peptides with simultaneous modulation of toxic activity against human red blood cells. AB - The toxicity of naturally occurring or designed antimicrobial peptides is a major barrier for converting them into drugs. To synthesize antimicrobial peptides with reduced toxicity, several amphipathic peptides were designed based on the leucine zipper sequence. The first one was a leucine zipper peptide (LZP); in others, leucine residues at the a- and/or d-position were substituted with single or double alanine residues. The results showed that LZP and its analogs exhibited appreciable and similar antibacterial activity against the tested gram-positive and gram-negative bacteria. However, the substitution of alanine progressively lowered the toxicity of LZP against human red blood cells (hRBCs). The substitution of leucine with alanine impaired the binding and localization of LZP to hRBCs, but had little effect on the peptide-induced damage of Escherichia coli cells. Although LZP and its analogs exhibited similar permeability, secondary structures, and localization in negatively charged membranes, significant differences were observed among these peptides in zwitterionic membranes. The results suggest a novel approach for designing antibacterial peptides with modulation of toxicity against hRBCs by employing the leucine zipper sequence. Also, to the best of our knowledge, the results demonstrate that this sequence could be utilized to design novel cell-selective molecules for the first time. PMID- 16717088 TI - Investigation of Cu(I)-dependent 2,4,5-trihydroxyphenylalanine quinone biogenesis in Hansenula polymorpha amine oxidase. AB - Copper, a mediator of redox chemistries in biology, is often found in enzymes that bind and reduce dioxygen. Among these, the copper amine oxidases catalyze the oxidative deamination of primary amines utilizing a type(II) copper center and 2,4,5-trihydroxyphenylalanine quinone (TPQ), a covalent cofactor derived from the post-translational modification of an active site tyrosine. Previous studies established the dependence of TPQ biogenesis on Cu(II); however, the dependence of cofactor formation on the biologically relevant Cu(I) ion has remained untested. In this study, we demonstrate that the apoform of the Hansenula polymorpha amine oxidase readily binds Cu(I) under anaerobic conditions and produces the quinone cofactor at a rate of 0.28 h(-1) upon subsequent aeration to yield a mature enzyme with kinetic properties identical to the protein product of the Cu(II)-dependent reaction. Because of the change in magnetic properties associated with the oxidation of copper, electron paramagnetic resonance spectroscopy was employed to investigate the nature of the rate-limiting step of Cu(I)-dependent cofactor biogenesis. Upon aeration of the unprocessed enzyme prebound with Cu(I), an axial Cu(II) electron paramagnetic resonance signal was found to appear at a rate equivalent to that for the cofactor. These data provide strong evidence for a rate-limiting release of superoxide from a Cu(II)(O(2)(.)) complex as a prerequisite for the activation of the precursor tyrosine and its transformation for TPQ. As copper is trafficked to intracellular protein targets in the reduced, Cu(I) state, these studies offer possible clues as to the physiological significance of the acquisition of Cu(I) by nascent H. polymorpha amine oxidase. PMID- 16717089 TI - Peptide aptamer-mediated inhibition of target proteins by sequestration into aggresomes. AB - Peptide aptamers (PAs) can be employed to block the intracellular function of target proteins. Little is known about the mechanism of PA-mediated protein inhibition. Here, we generated PAs that specifically bound to the duck hepatitis B virus (HBV) core protein. Among them, PA34 strongly blocked duck HBV replication by inhibiting viral capsid formation. We found that PA34 led to a dramatic intracellular redistribution of its target protein into perinuclear inclusion bodies, which exhibit the typical characteristics of aggresomes. As a result, the core protein is efficiently removed from the viral life cycle. Corresponding findings were obtained for bioactive PAs that bind to the HBV core protein or to the human papillomavirus-16 (HPV16) E6 protein, respectively. The observation that PAs induce the specific sequestration of bound proteins into aggresomes defines a novel mechanism as to how this new class of intracellular inhibitors blocks the function of their target proteins. PMID- 16717090 TI - Double-stranded RNA-dependent protein kinase phosphorylation of the alpha-subunit of eukaryotic translation initiation factor 2 mediates apoptosis. AB - As the molecular processes of complex cell stress signaling pathways are defined, the subsequent challenge is to elucidate how each individual event influences the final biological outcome. Phosphorylation of the translation initiation factor 2 (eIF2alpha)atSer(51) is a molecular signal that inhibits translation in response to activation of any of four diverse eIF2alpha stress kinases. We used gene targeting to replace the wild-type Ser(51) allele with an Ala in the eIF2alpha gene to test the hypothesis that translational control through eIF2alpha phosphorylation is a central death stimulus in eukaryotic cells. Homozygous eIF2alpha mutant mouse embryo fibroblasts were resistant to the apoptotic effects of dsRNA, tumor necrosis factor-alpha, and serum deprivation. TNFalpha treatment induced eIF2alpha phosphorylation and activation of caspase 3 primarily through the dsRNA-activated eIF2alpha kinase PKR. In addition, expression of a phospho mimetic Ser(51) to Asp mutant eIF2alpha-activated caspase 3, indicating that eIF2alpha phosphorylation is sufficient to induce apoptosis. The proapoptotic effects of PKR-mediated eIF2alpha phosphorylation contrast with the anti apoptotic response upon activation of the PKR-related endoplasmic reticulum eIF2alpha kinase, PERK. Therefore, divergent fates of death and survival can be mediated through phosphorylation at the same site within eIF2alpha. We propose that eIF2alpha phosphorylation is fundamentally a death signal, yet it may promote either death or survival, depending upon coincident signaling events. PMID- 16717091 TI - The polycomb group protein EZH2 is required for mammalian circadian clock function. AB - We examined the importance of histone methylation by the polycomb group proteins in the mouse circadian clock mechanism. Endogenous EZH2, a polycomb group enzyme that methylates lysine 27 on histone H3, co-immunoprecipitates with CLOCK and BMAL1 throughout the circadian cycle in liver nuclear extracts. Chromatin immunoprecipitation revealed EZH2 binding and di- and trimethylation of H3K27 on both the Period 1 and Period 2 promoters. A role of EZH2 in cryptochrome-mediated transcriptional repression of the clockwork was supported by overexpression and RNA interference studies. Serum-induced circadian rhythms in NIH 3T3 cells in culture were disrupted by transfection of an RNA interfering sequence targeting EZH2. These results indicate that EZH2 is important for the maintenance of circadian rhythms and extend the activity of the polycomb group proteins to the core clockwork mechanism of mammals. PMID- 16717092 TI - Structure of the p53 core domain dimer bound to DNA. AB - The p53 tumor suppressor protein binds to DNA as a dimer of dimers to regulate transcription of genes that mediate responses to cellular stress. We have prepared a cross-linked trapped p53 core domain dimer bound to decamer DNA and have determined its structure by x-ray crystallography to 2.3A resolution. The p53 core domain subunits bind nearly symmetrically to opposite faces of the DNA in a head-to-head fashion with a loophelix motif making sequence-specific DNA contacts and bending the DNA by about 20 degrees at the site of protein dimerization. Protein subunit interactions occur over the central DNA minor groove and involve residues from a zinc-binding region. Analysis of tumor derived p53 mutations reveals that the dimerization interface represents a third hot spot for mutation that also includes residues associated with DNA contact and protein stability. Residues associated with p53 dimer formation on DNA are poorly conserved in the p63 and p73 paralogs, possibly contributing to their functional differences. We have used the dimeric protein-DNA complex to model a dimer of p53 dimers bound to icosamer DNA that is consistent with solution bending data and suggests that p53 core domain dimer-dimer contacts are less frequently mutated in human cancer than intra-dimer contacts. PMID- 16717093 TI - The importance of P-loop and domain movements in EF-Tu for guanine nucleotide exchange. AB - Elongation factor Ts (EF-Ts) is the guanine nucleotide exchange factor for elongation factor Tu (EF-Tu). An important feature of the nucleotide exchange is the structural rearrangement of EF-Tu in the EF-Tu.EF-Ts complex caused by insertion of Phe-81 of EF-Ts between His-84 and His-118 of EF-Tu. In this study, the contribution of His-118 to nucleotide release was studied by pre-steady state kinetic analysis of nucleotide exchange in EF-Tu mutants in which His-118 was replaced by Ala or Glu. Intrinsic as well as EF-Ts-catalyzed release of GDP/GTP was affected by the mutations, resulting in an approximately 10-fold faster spontaneous nucleotide release and a 10-50-fold slower EF-Ts-catalyzed nucleotide release. The effects are attributed to the interference of the mutations with the EF-Ts-induced movements of the P-loop of EF-Tu and changes at the domain 1/3 interface, leading to the release of the beta-phosphate group of GTP/GDP. The K(d) for GTP is increased by more than 40 times when His-118 is replaced with Glu, which may explain the inhibition by His-118 mutations of aminoacyl-tRNA binding to EF-Tu. The mutations had no effect on EF-Tu-dependent delivery of aminoacyl-tRNA to the ribosome. PMID- 16717095 TI - Mechanism of assembly of G protein betagamma subunits by protein kinase CK2 phosphorylated phosducin-like protein and the cytosolic chaperonin complex. AB - Phosducin-like protein (PhLP) is a widely expressed binding partner of the G protein betagamma subunit complex (Gbetagamma) that has been recently shown to catalyze the formation of the Gbetagamma dimer from its nascent polypeptides. Phosphorylation of PhLP at one or more of three consecutive serines (Ser-18, Ser 19, and Ser-20) is necessary for Gbetagamma dimer formation and is believed to be mediated by the protein kinase CK2. Moreover, several lines of evidence suggest that the cytosolic chaperonin complex (CCT) may work in concert with PhLP in the Gbetagamma-assembly process. The results reported here delineate a mechanism for Gbetagamma assembly in which a stable ternary complex is formed between PhLP, the nascent Gbeta subunit, and CCT that does not include Ggamma. PhLP phosphorylation permits the release of a PhLP x Gbeta intermediate from CCT, allowing Ggamma to associate with Gbeta in this intermediate complex. Subsequent interaction of Gbetagamma with membranes releases PhLP for another round of assembly. PMID- 16717094 TI - The mechanism of direct heme transfer from the streptococcal cell surface protein Shp to HtsA of the HtsABC transporter. AB - The heme-binding proteins Shp and HtsA are part of the heme acquisition machinery found in Streptococcus pyogenes. The hexacoordinate heme (Fe(II)-protoporphyrin IX) or hemochrome form of holoShp (hemoShp) is stable in air in Tris-HCl buffer, pH 8.0, binds to apoHtsA with a K(d) of 120 +/- 18 microm, and transfers its heme to apoHtsA with a rate constant of 28 +/- 6s(-1) at 25 degrees C, pH 8.0. The hemoHtsA product then autoxidizes to the hexacoordinate hemin (Fe(III) protoporphyrin IX) or hemichrome form (hemiHtsA) with an apparent rate constant of 0.017 +/- 0.002 s(-1). HemiShp also rapidly transfers hemin to apoHtsA through a hemiShp.apoHtsA complex (K(d) = 48 +/- 7 microM) at a rate approximately 40,000 times greater than the rate of simple hemin dissociation from hemiShp into solvent (k(transfer) = 43 +/- 3s(-1) versus k(-hemin) = 0.0003 +/- 0.00006 s( 1)). The rate constants for hemin binding to and dissociation from HtsA (k'(hemin) approximately 80 microm(-1) s(-1), k(-hemin) = 0.0026 +/- 0.0002 s( 1)) are 50- and 10-fold greater than the corresponding rate constants for Shp (k(hemin) approximately 1.6 microM(-1) s(-1), k(-hemin) = 0.0003 s(-1)), which implies that HtsA has a more accessible active site. However, the affinity of apoHtsA for hemin (k(hemin) approximately 31,000 microm(-1)) is roughly 5-fold greater than that of apoShp (k(hemin) approximately 5,300 microM(-1)), accounting for the net transfer from Shp to HstA. These results support a direct, rapid, and affinity-driven mechanism of heme and hemin transfer from the cell surface receptor Shp to the ATP-binding cassette transporter system. PMID- 16717096 TI - Phosphorylation of viral RNA-dependent RNA polymerase and its role in replication of a plus-strand RNA virus. AB - Central to the process of plus-strand RNA virus genome amplification is the viral RNA-dependent RNA polymerase (RdRp). Understanding its regulation is of great importance given its essential function in viral replication and the common architecture and catalytic mechanism of polymerases. Here we show that Turnip yellow mosaic virus (TYMV) RdRp is phosphorylated, when expressed both individually and in the context of viral infection. Using a comprehensive biochemical approach, including metabolic labeling and mass spectrometry analyses, phosphorylation sites were mapped within an N-terminal PEST sequence and within the highly conserved palm subdomain of RNA polymerases. Systematic mutational analysis of the corresponding residues in a reverse genetic system demonstrated their importance for TYMV infectivity. Upon mutation of the phosphorylation sites, distinct steps of the viral cycle appeared affected, but in contrast to other plus-strand RNA viruses, the interaction between viral replication proteins was unaltered. Our results also highlighted the role of another TYMV-encoded replication protein as an antagonistic protein that may prevent the inhibitory effect of RdRp phosphorylation on viral infectivity. Based on these data, we propose that phosphorylation-dependent regulatory mechanisms are essential for viral RdRp function and virus replication. PMID- 16717097 TI - Insulin reverses growth hormone-induced homologous desensitization. AB - Growth hormone (GH) is secreted in a pulsatile pattern to promote body growth and metabolism. GH exerts its function by activating several signaling pathways, including JAK2/STAT and MEK/ERK. ERK1/2 activation by GH plays important roles in gene expression, cell proliferation, and growth. We previously reported that in rat H4IIE hepatoma cells after an initial GH exposure, a second GH exposure induces STAT5 phosphorylation but not ERK1/2 phosphorylation (Ji, S., Frank, S. J., and Messina, J. L. (2002) J. Biol. Chem. 277, 28384-28393). In this study the mechanisms underlying GH-induced homologous desensitization were investigated. A second GH exposure activated the signaling intermediates upstream of MEK/ERK, including JAK2, Ras, and Raf-1. This correlated with recovery of GH receptor levels, but was insufficient for GH-induced phosphorylation of MEK1/2 and ERK1/2. Insulin restored the ability of a second GH exposure to induce phosphorylation of MEK1/2 and ERK1/2 without altering GH receptor levels or GH-induced phosphorylation/activation of JAK2 and Raf-1. GH and insulin synergized in promoting cell proliferation. Further investigation suggested that insulin increased the amount of MEK bound to KSR (kinase suppressor of Ras) and restored GH-induced tyrosine phosphorylation of KSR. Previous GH exposure also induced desensitization of STAT1 and STAT3 phosphorylation, but this desensitization was not reversed by insulin. Thus, insulin-regulated resensitization of GH signaling may be necessary to reset the complete response to GH after a normal, physiologic pulse of GH. PMID- 16717098 TI - The oxygen transport system in three species of the boreal fish family Gadidae. Molecular phylogeny of hemoglobin. AB - The Arctic and Antarctic marine faunas differ by age and isolation. Fishes of the two polar regions have undergone different regional histories that have driven the physiological diversities. Antarctic fish are highly stenothermal, in keeping with stable water temperatures, whereas Arctic fish, being exposed to seasonal temperature variations, exhibit higher physiological plasticity. This study reports the characterization of the oxygen transport system of three Arctic species of the family Gadidae, namely the Arctic cod Arctogadus glacialis, the polar cod Boreogadus saida, and the Atlantic cod Gadus morhua. Unlike Antarctic notothenioids, the blood displays high multiplicity, i.e. it has three hemoglobins, similar to many other acanthomorph teleosts. In the most abundant hemoglobin, oxygen binding is modulated by heterotropic effectors, with marked Bohr and Root effects. Remarkably, in two species (A. glacialis and B. saida), the Hill coefficient is very close to one in the whole pH range, indicating the apparent absence of cooperativity. The amino acid sequences have been used to gain insight into the evolution history of globins of polar fish. The results indicate that Arctic and Antarctic globins have different phylogenies and lead us to suggest that the selective pressure of environment stability allows the phylogenetic signal to be maintained in the Antarctic sequences, whereas environmental variability would tend to disrupt this signal in the Gadidae sequences. PMID- 16717099 TI - A screen for genes of heme uptake identifies the FLC family required for import of FAD into the endoplasmic reticulum. AB - Although Candida albicans and Saccharomyces cerevisiae express very similar systems of iron uptake, these species differ in their capacity to use heme as a nutritional iron source. Whereas C. albicans efficiently takes up heme, S. cerevisiae grows poorly on media containing heme as the sole source of iron. We identified a gene from C. albicans that would enhance heme uptake when expressed in S. cerevisiae. Overexpression of CaFLC1 (for flavin carrier 1) stimulated the growth of S. cerevisiae on media containing heme iron. In C. albicans, deletion of both alleles of CaFLC1 resulted in a decrease in heme uptake activity, whereas overexpression of CaFLC1 resulted in an increase in heme uptake. The S. cerevisiae genome contains three genes with homology to CaFLC1, and two of these, termed FLC1 and FLC2, also stimulated growth on heme when overexpressed in S. cerevisiae. The S. cerevisiae Flc proteins were detected in the endoplasmic reticulum and the FLC genes encoded an essential function, as strains deleted for either FLC1 or FLC2 were viable, but deletion of both FLC1 and FLC2 was synthetically lethal. FLC gene deletion resulted in pleiotropic phenotypes related to defects in cell wall integrity. High copy suppressors of this synthetic lethality included three mannosyltransferases, VAN1, KTR4, and HOC1. FLC deletion strains exhibited loss of cell wall mannose phosphates, defects in cell wall assembly, and delayed maturation of carboxypeptidase Y. Permeabilized cells lacking FLC proteins exhibited dramatic loss of FAD import activity. We propose that the FLC genes are required for import of FAD into the lumen of the endoplasmic reticulum, where it is required for disulfide bond formation. PMID- 16717100 TI - Thyroid hormone stimulates protein synthesis in the cardiomyocyte by activating the Akt-mTOR and p70S6K pathways. AB - Thyroid hormones affect cardiac growth and phenotype; however, the mechanisms by which the hormones induce cardiomyocyte hypertrophy remain uncharacterized. Tri iodo-L-thyronine (T3) treatment of cultured cardiomyocytes for 24 h resulted in a 41 +/- 5% (p < 0.001) increase in [(3)H]leucine incorporation into total cellular protein. This response was abrogated by the phosphatidylinositol 3-kinase (PI3K) inhibitor, wortmannin. Co-immunoprecipitation studies showed a direct interaction of cytosol-localized thyroid hormone receptor TRalpha1 and the p85alpha subunit of PI3K. T3 treatment rapidly increased PI3K activity by 52 +/- 3% (p < 0.005), which resulted in increased phosphorylation of downstream kinases Akt and mammalian target of rapamycin (mTOR). This effect was abrogated by pretreatment with wortmannin or LY294002. Phosphorylation of p70(S6K), a known target of mTOR, occurred rapidly following T3 treatment and was inhibited by rapamycin and wortmannin. In contrast, phosphorylation of the p85 variant of S6K in response to T3 was not blocked by LY294002, wortmannin, or rapamycin, thus supporting a T3 activated pathway independent of PI3K and mTOR. 40 S ribosomal protein S6, a target of p70(S6K), and 4E-BP1, a target of mTOR, were both phosphorylated within 15-25 min of T3 treatment and could be inhibited by wortmannin and rapamycin. Thus, rapid T3-mediated activation of PI3K by cytosolic TRalpha1 and subsequent activation of the Akt-mTOR-S6K signaling pathway may underlie one of the mechanisms by which thyroid hormone regulates physiological cardiac growth. PMID- 16717101 TI - Domain structure and protein interactions of the silent information regulator Sir3 revealed by screening a nested deletion library of protein fragments. AB - Transcriptional silencing in yeast is mediated by the interactions of silent information regulator (Sir) proteins with chromatin and with one another. The stable association of Sir3 with Sir4 is mediated by a C-terminal region of Sir3 that has additional functions including the dimerization of Sir3. We have developed a simple, robust expression screening methodology that allows for the unbiased identification of functional protein domains expressed from nested deletion libraries of full-length genes. Using these methodologies, Sir3 dimerization was shown to be mediated by two separate domains. One of these domains also binds cooperatively to the C-terminal coiled-coil motif of Sir4 and dimerization further increases the affinity of Sir3 for Sir4. The resulting Sir3 Sir4 complexes form progressively higher order assemblies with increasing protein concentration, with implications for the mechanism of gene silencing. PMID- 16717102 TI - G1/S arrest induced by histone deacetylase inhibitor sodium butyrate in E1A + Ras transformed cells is mediated through down-regulation of E2F activity and stabilization of beta-catenin. AB - Tumor cells are often characterized by a high and growth factor-independent proliferation rate. We have previously shown that REF cells transformed with oncogenes E1A and c-Ha-Ras do not undergo G(1)/S arrest of the cell cycle after treatment with genotoxic factors. In this work, we used sodium butyrate, a histone deacetylase inhibitor, to show that E1A + Ras transformants were able to stop proliferation and undergo G(1)/S arrest. Apart from inducing G(1)/S arrest, sodium butyrate was shown to change expression of a number of cell cycle regulatory genes. It down-regulated cyclins D1, E, and A as well as c-myc and cdc25A and up-regulated the cyclin-kinase inhibitor p21(waf1). Accordingly, activities of cyclin E-Cdk2 and cyclin A-Cdk2 complexes in sodium butyrate treated cells were decreased substantially. Strikingly, E2F1 expression was also down-modulated at the levels of gene transcription, the protein content, and the E2F transactivating capability. To further study the role of p21(waf1) in the sodium butyrate-induced G(1)/S arrest and the E2F1 down-modulation, we established E1A + Ras transformants from mouse embryo fibroblast cells with deletion of the cdkn1a (p21(waf1)) gene. Despite the absence of p21(waf1), sodium butyrate-treated mERas transformants reveal a slightly delayed G(1)/S arrest as well as down-modulation of E2F1 activity, implying that the observed effects are mediated through an alternative p21(waf1)-independent signaling pathway. Subsequent analysis showed that sodium butyrate induced accumulation of beta catenin, a downstream component of the Wnt signaling. The results obtained indicate that the antiproliferative effect of histone deacetylase inhibitors on E1A + Ras-transformed cells can be mediated, alongside other mechanisms, through down-regulation of E2F activity and stabilization of beta-catenin. PMID- 16717103 TI - Detection of sulfide release from the oxygen-sensing [4Fe-4S] cluster of FNR. AB - The Escherichia coli FNR protein regulates the transcription of >100 genes in response to environmental O2, thereby coordinating the response to anoxia. Under O2-limiting conditions, FNR binds a [4Fe-4S]2+ cluster through four cysteine residues (Cys20, Cys23, Cys29, Cys122). The acquisition of the [4Fe-4S]2+ cluster converts FNR into the transcriptionally active dimeric form. Upon exposure to O2, the cluster converts to a [2Fe-2S]2+ form, generating FNR monomers that no longer bind DNA with high affinity. The mechanism of the cluster conversion reaction and the nature of the released iron and sulfur are of considerable current interest. Here, we report the application of a novel in vitro method, involving 5,5' dithiobis-(2-nitrobenzoic acid), for determining the oxidation state of the sulfur atoms released during FNR cluster conversion following the addition of O2. Conversion of [4Fe-4S]2+ to [2Fe-2S]2+ clusters by O2 for both native and reconstituted FNR results in the release of approximately 2 sulfide ions per [4Fe 4S]2+ cluster. This demonstrates that the reaction between O2 and the [4Fe-4S]2+ cluster does not require sulfide oxidation and hence must entail iron oxidation. PMID- 16717104 TI - GlycoPEGylation of recombinant therapeutic proteins produced in Escherichia coli. AB - Covalent attachment of polyethylene glycol, PEGylation, has been shown to prolong the half-life and enhance the pharmacodynamics of therapeutic proteins. Current methods for PEGylation, which rely on chemical conjugation through reactive groups on amino acids, often generate isoforms in which PEG is attached at sites that interfere with bioactivity. Here, we present a novel strategy for site directed PEGylation using glycosyltransferases to attach PEG to O-glycans. The process involves enzymatic GalNAc glycosylation at specific serine and threonine residues in proteins expressed without glycosylation in Escherichia coli, followed by enzymatic transfer of sialic acid conjugated with PEG to the introduced GalNAc residues. The strategy was applied to three therapeutic polypeptides, granulocyte colony stimulating factor (G-CSF), interferon-alpha2b (IFN-alpha2b), and granulocyte/macrophage colony stimulating factor (GM-CSF), which are currently in clinical use. PMID- 16717105 TI - Beneficial effects of metolazone in a rat model of preeclampsia. AB - Preeclampsia is a disorder that continues to exact a significant toll with respect to maternal morbidity and mortality as well as fetal wastage. Furthermore, the treatment of this disorder has not changed significantly in 50 years and is unsatisfactory. The use of diuretics in this syndrome is controversial because there is a concern related to potential baleful effects of volume contraction leading to a possible further decrement in the perfusion of the maternal-fetal unit. Metolazone is a diuretic/antihypertensive agent, which has a therapeutic effect on blood pressure (BP) in human essential hypertension without causing a natriuresis. We administered the drug in nondiuretic doses in a rat model of preeclampsia previously developed in this laboratory. The drug reduced BP without an accompanying natriuresis. Although there was a trend toward an improvement in intrauterine growth restriction, as determined by litter size and the number of pups demonstrating malformations, the values did not reach statistical significance. We conclude that metolazone, in low dosage, is an effective antihypertensive in this rat model. These studies have implications for the treatment of the human disorder. PMID- 16717106 TI - L-arginine chlorination results in the formation of a nonselective nitric-oxide synthase inhibitor. AB - Reduced nitric oxide (NO) bioavailability and impaired vascular function are the key pathological characteristics of inflammatory diseases such as atherosclerosis. We have recently found that leukocyte-derived hypochlorous acid is able to react with the nitric-oxide synthase (NOS) substrate L-arginine to produce chlorinated L-arginine (cl-L-Arg). Interestingly, cl-L-Arg potently inhibits the formation of NO metabolites in cultured endothelial cells. It is unknown whether cl-L-Arg has a direct inhibitory effect on endothelial NOS (eNOS). In addition, the effect of cl-L-Arg on the other NOS isoforms, neuronal NOS (nNOS) and inducible NOS (iNOS), is also unknown. Therefore, we designed the current study to test the effects of cl-L-Arg on eNOS, nNOS, and iNOS. Using recombinant NOS, we found that cl-L-Arg had a direct inhibitory effect on the activity of NOS. The effect of cl-L-Arg on NOS activity is nonselective, as all three NOS isoforms were inhibited with a similar IC(50). We further determined the effect of cl-L-Arg on the three NOS isoforms at the tissue level. The results demonstrated that cl-L-Arg potently inhibited all three NOS isoform-mediated vessel reactivities, as well as the NOS signaling molecule cGMP. Cl-L-Arg might serve as a novel endogenous NOS inhibitor and an important mediator for vascular dysfunction under inflammatory conditions such as atherosclerosis. Blocking cl-L Arg formation may be a new therapeutic approach to cardiovascular diseases. PMID- 16717107 TI - Collagenase-2 and -3 mediate epidermal growth factor receptor transactivation by bradykinin B2 receptor in kidney cells. AB - We have previously shown that stimulation of extracellular signal-regulated protein kinase (ERK) by bradykinin (BK) in murine inner medullary collecting duct (mIMCD)-3 cells is mediated by epidermal growth factor receptor (EGFR) transactivation. The mechanism of EGFR transactivation seemed to be novel, because it does not require phospholipase C, Ca(2+), calmodulin, protein kinase C, G alpha(i) subunits, or EGFR-B(2) receptor heterodimerization. In this study, we demonstrated the involvement of matrix metalloproteinases (MMPs) in B(2) receptor-induced EGFR transactivation using their broad-spectrum inhibitors batimastat and N-[(2R)-2-(hydroxamidocarbonylmethyl)-4-methylpentanoyl]-l tryptophan methylamide (Galardin) (GM-6001). Selective inhibitors for collagenase 2 and -3 (MMP-8 and MMP-13, respectively) blocked BK-induced EGFR phosphorylation and ERK activation, whereas inhibitors for MMP-1, -2, -3, -7, or -9 were without effect. Transfection of mIMCD-3 cells with MMP-8 small interfering RNA (siRNA) resulted in approximately 50% decrease of BK-induced ERK activation. A neutralizing antibody against MMP-13 as well as transfection with MMP-13 siRNA produced a similar effect. Inhibition of both collagenases resulted in approximately 65% decrease of BK-induced ERK activation, supporting roles for both enzymes. Stimulation of mIMCD-3 cells with 10 nM BK increased the activity of collagenases in concentrated culture media within 10 min. Moreover, recombinant MMP-13 and MMP-8, when applied to mIMCD-3 cells for 10 min without BK, stimulated tyrosine phosphorylation of EGFR and caused approximately 250% increase over basal ERK phosphorylation comparable with BK-induced ERK activation. Collagenases-induced ERK activation was inhibited by 4-(3 chloroanilino)-6,7-dimethoxyquinazoline (AG-1478) and thus dependent on EGFR tyrosine kinase activity. This study demonstrates a novel role for collagenase-2 and -3 in signaling of the G(q)-coupled BK B(2) receptor in mIMCD-3 cells. PMID- 16717108 TI - Elimination of Cerenkov interference in a fibre-optic-coupled radiation dosemeter. AB - An optical fibre point dosemeter based on the gated detection of the luminescence from a Cu(1+)-doped fused quartz detector effectively eliminated errors due to Cerenkov radiation and native fibre fluorescence. The gated optical fibre dosemeter overcomes serious problems faced by scintillation and optically stimulated luminescence approaches to optical fibre point dosimetry. The dosemeter was tested using an external beam radiotherapy machine that provided pulses of 6 MV X rays. Gated detection was used to discriminate the signal collected during the radiation pulses, which included contributions from Cerenkov radiation and native fibre fluorescence, from the signal collected between the radiation pulses, which contained only the long-lived luminescence from the Cu(1+)-doped fused quartz detector. Gated detection of the luminescence provided accurate, real-time dose measurements that were linear with absorbed dose, independent of dose rate and that were accurate for all field sizes studied. PMID- 16717110 TI - EPR properties of intact and deproteinated dentin. AB - The effect of sample preparation on dentin electron paramagnetic resonance (EPR) spectra was investigated. The analysis was performed on dentin samples prepared by pure mechanical treatment or by an alkaline deproteination method. It was observed that (1) mechanical treatment induces both stable and transient signals, depending on the specific mechanical operation applied; and (2) sodium hydroxide deproteination removes the native signal, but at the same time introduces new confounding signals in the EPR spectrum. Also, it increases the radiation sensitivity of dentin. These findings suggest that attention must be paid to dentin sample preparation. PMID- 16717109 TI - Novel shielding materials for space and air travel. AB - The reduction of dose onboard spacecraft and aircraft by appropriate shielding measures plays an essential role in the future development of space exploration and air travel. The design of novel shielding strategies and materials may involve hydrogenous composites, as it is well known that liquid hydrogen is most effective in attenuating charged particle radiation. As precursor for a later flight experiment, the shielding properties of newly developed hydrogen-rich polymers and rare earth-doped high-density rubber were tested in various ground based neutron and heavy ion fields and compared with aluminium and polyethylene as reference materials. Absorbed dose, average linear energy transfer and gamma equivalent neutron absorbed dose were determined by means of LiF:Mg,Ti thermoluminescence dosemeters and CR-39 plastic nuclear track detectors. First results for samples of equal aerial density indicate that selected hydrogen-rich plastics and rare-earth-doped rubber may be more effective in attenuating cosmic rays by up to 10% compared with conventional aluminium shielding. The appropriate adaptation of shielding thicknesses may thus allow reducing the biologically relevant dose. Owing to the lower density of the plastic composites, mass savings shall result in a significant reduction of launch costs. The experiment was flown as part of the European Space Agency's Biopan-5 mission in May 2005. PMID- 16717111 TI - A comprehensive comparative study of the predose effect for three quartz crystals of different origin. AB - The study of the thermoluminiscence (TL) sensitivity of quartz due to heat and irradiation treatments is of importance in dating and retrospective dosimetry. A comprehensive comparative study of the predose effect was carried out for three types of quartz of different origin. Complete TL vs. dose and sensitivity S vs. predose curves were obtained for the dose range of 0.1 < D < 400 Gy. Additional complete sensitivity vs. predose curves were obtained for samples which underwent a combined predose irradiation and a subsequent heat treatment to 500 degrees C. Although the TL vs. dose curves showed very different behaviours, the sensitivity vs. predose curves showed several common characteristics. The sensitivity vs. predose curves showed abrupt changes approximately 10 Gy. The sensitivity after a combined predose irradiation and heat treatment to 500 degrees C showed a very gradual change in the whole dose range studied. These results are explained qualitatively by using the modified Zimmerman model for quartz. PMID- 16717112 TI - Study of an improved Allyl Di-Glycol carbonate sheet for high energy proton detection. AB - An allyl di-glycol carbonate (ADC) sheet which has been utilised as a neutron detector for personal dosimetry has recently been studied for its application as a device for radiation exposure control for astronauts in space, where protons are the dominant radiation. It is known that the fabrication process, modified by adding some kind of antioxidant to improve the sensitivity of ADC to high energy protons, causes a substantial increase in false tracks, which disturb the automatic counting of proton tracks using the auto-image analyser. This made clear the difficulty of fabricating ADC sheets which have sufficient sensitivity to high energy protons, while maintaining a good surface. In this study, we have tried to modify the fabrication process to improve the sensitivity to high energy protons without causing a deterioration of the surface condition of ADC sheets. We have successfully created fairly good products. PMID- 16717113 TI - Autoimmune arthritis associated with mutated interleukin (IL)-6 receptor gp130 is driven by STAT3/IL-7-dependent homeostatic proliferation of CD4+ T cells. AB - Mice homozygous for the F759 mutation in the gp130 interleukin (IL)-6 receptor subunit have enhanced gp130-mediated signal transducer and activator of transcription (STAT)3 activation and spontaneously developed a lymphocyte mediated rheumatoid arthritis-like joint disease. Here, we show that the development of the disease is dependent on both major histocompatibility complex (MHC) II-restricted CD4+ T cells and IL-6 family cytokines. In spite of the necessity for CD4+ T cells, the gp130 mutation was only required in nonhemtopoietic cells for the disease. The gp130 mutation resulted in enhanced production of IL-7. Conditional knockout of STAT3 in nonlymphoid cells showed that the enhancement of IL-7 production was dependent on STAT3 activation by IL-6 family cytokines. Homeostatic proliferation of CD4+ T cells was enhanced in gp130 mutant mice and acceleration of homeostatic proliferation enhanced the disease, whereas the inhibition of homeostatic proliferation suppressed the disease. Anti IL-7 antibody treatment inhibited not only the enhanced homeostatic proliferation, but also the disease in gp130 mutant mice. Thus, our results show that autoimmune disease in gp130 mutant mice is caused by increased homeostatic proliferation of CD4+ T cells, which is due to elevated production of IL-7 by nonhematopoietic cells as a result of IL-6 family cytokine-gp130-STAT3 signaling. PMID- 16717115 TI - T helper type 1-specific Brg1 recruitment and remodeling of nucleosomes positioned at the IFN-gamma promoter are Stat4 dependent. AB - Transcriptional competence of the interferon-gamma (IFN-gamma) locus is enhanced as Th1 effectors develop from naive CD4 T lymphocytes; conversely, this gene is repressed during Th2 differentiation. We now show that the Switch (Swi)-sucrose nonfermenter (SNF) component Brahma-related gene 1 (Brg1) is recruited, and positioned nucleosomes are remodeled, in a Th1-specific manner that is dependent on the transcription factor Stat4 and calcineurin phosphatase activity. Interference with specific components of mammalian Swi-SNF complexes decreased CD4 T cell differentiation into IFN-gamma-positive Th1 cells. These findings reveal a collaborative mechanism of IFN-gamma gene regulation during Th1 differentiation and suggest that a Th1-specific chromatin structure is created by early recruitment of Swi-SNF complexes and nucleosome remodeling dependent on Stat4 and calcineurin activation. PMID- 16717114 TI - Impaired degranulation but enhanced cytokine production after Fc epsilonRI stimulation of diacylglycerol kinase zeta-deficient mast cells. AB - Calcium and diacylglycerol are critical second messengers that together effect mast cell degranulation after allergen cross-linking of immunoglobulin (Ig)E bound FcepsilonRI. Diacylglycerol kinase (DGK)zeta is a negative regulator of diacylglycerol-dependent signaling that acts by converting diacylglycerol to phosphatidic acid. We reported previously that DGKzeta-/- mice have enhanced in vivo T cell function. Here, we demonstrate that these mice have diminished in vivo mast cell function, as revealed by impaired local anaphylactic responses. Concordantly, DGKzeta-/- bone marrow-derived mast cells (BMMCs) demonstrate impaired degranulation after Fc epsilonRI cross-linking, associated with diminished phospholipase Cgamma activity, calcium flux, and protein kinase C betaII membrane recruitment. In contrast, Ras-Erk signals and interleukin-6 production are enhanced, both during IgE sensitization and after antigen cross linking of Fc epsilonRI. Our data demonstrate dissociation between cytokine production and degranulation in mast cells and reveal the importance of DGK activity during IgE sensitization for proper attenuation of Fc epsilonRI signals. PMID- 16717117 TI - Role of the caspase-1 inflammasome in Salmonella typhimurium pathogenesis. AB - Caspase-1 is activated by a variety of stimuli after the assembly of the "inflammasome," an activating platform made up of a complex of the NOD-LRR family of proteins. Caspase-1 is required for the secretion of proinflammatory cytokines, such as interleukin (IL)-1beta and IL-18, and is involved in the control of many bacterial infections. Paradoxically, however, its absence has been reported to confer resistance to oral infection by Salmonella typhimurium. We show here that absence of caspase-1 or components of the inflammasome does not result in resistance to oral infection by S. typhimurium, but rather, leads to increased susceptibility to infection. PMID- 16717116 TI - A cyanobacterial LPS antagonist prevents endotoxin shock and blocks sustained TLR4 stimulation required for cytokine expression. AB - Toll-like receptors (TLRs) function as primary sensors that elicit coordinated innate immune defenses through recognition of microbial products and induction of immune and proinflammatory genes. Here we report the identification and biological characterization of a lipopolysaccharide (LPS)-like molecule extracted from the cyanobacterium Oscillatoria Planktothrix FP1 (cyanobacterial product [CyP]) that is not stimulatory per se but acts as a potent and selective antagonist of bacterial LPS. CyP binds to MD-2 and efficiently competes with LPS for binding to the TLR4-MD-2 receptor complex. The addition of CyP together with LPS completely inhibited both MyD88- and TRIF-dependent pathways and suppressed the whole LPS-induced gene transcription program in human dendritic cells (DCs). CyP protected mice from endotoxin shock in spite of a lower capacity to inhibit LPS stimulation of mouse DCs. Interestingly, the delayed addition of CyP to DCs responding to LPS strongly inhibited signaling and cytokine production by immediate down-regulation of inflammatory cytokine mRNAs while not affecting other aspects of DC maturation, such as expression of major histocompatibility complex molecules, costimulatory molecules, and CCR7. Collectively, these results indicate that CyP is a potent competitive inhibitor of LPS in vitro and in vivo and reveal the requirement of sustained TLR4 stimulation for induction of cytokine genes in human DCs. PMID- 16717119 TI - IFNgamma-dependent, spontaneous development of colorectal carcinomas in SOCS1 deficient mice. AB - Approximately 20% of human cancers are estimated to develop from chronic inflammation. Recently, the NF-kappaB pathway was shown to play an essential role in promoting inflammation-associated cancer, but the role of the JAK/STAT pathway, another important signaling pathway of proinflammatory cytokines, remains to be investigated. Suppressor of cytokine signaling-1 (SOCS1) acts as an important physiological regulator of cytokine responses, and silencing of the SOCS1 gene by DNA methylation has been found in several human cancers. Here, we demonstrated that SOCS1-deficient mice (SOCS1-/- Tg mice), in which SOCS1 expression was restored in T and B cells on a SOCS1-/- background, spontaneously developed colorectal carcinomas carrying nuclear beta-catenin accumulation and p53 mutations at 6 months of age. However, interferon (IFN)gamma-/- SOCS1-/- mice and SOCS1-/- Tg mice treated with anti-IFNgamma antibody did not develop such tumors. STAT3 and NF-kappaB activation was evident in SOCS1-/- Tg mice, but these were not sufficient for tumor development because these are also activated in IFNgamma-/- SOCS1-/- mice. However, colons of SOCS1-/- Tg mice, but not IFNgamma /- SOCS1-/- mice, showed hyperactivation of STAT1, which resulted in the induction of carcinogenesis-related enzymes, cyclooxygenase-2 and inducible nitric oxide synthase. These data strongly suggest that SOCS1 is a unique antioncogene which prevents chronic inflammation-mediated carcinogenesis by regulation of the IFNgamma/STAT1 pathways. PMID- 16717118 TI - An Ixodes scapularis protein required for survival of Anaplasma phagocytophilum in tick salivary glands. AB - Anaplasma phagocytophilum is the agent of human anaplasmosis, the second most common tick-borne illness in the United States. This pathogen, which is closely related to obligate intracellular organisms in the genera Rickettsia, Ehrlichia, and Anaplasma, persists in ticks and mammalian hosts; however, the mechanisms for survival in the arthropod are not known. We now show that A. phagocytophilum induces expression of the Ixodes scapularis salp16 gene in the arthropod salivary glands during vector engorgement. RNA interference-mediated silencing of salp16 gene expression interfered with the survival of A. phagocytophilum that entered ticks fed on A. phagocytophilum-infected mice. A. phagocytophilum migrated normally from A. phagocytophilum-infected mice to the gut of engorging salp16 deficient ticks, but up to 90% of the bacteria that entered the ticks were not able to successfully infect I. scapularis salivary glands. These data demonstrate the specific requirement of a pathogen for a tick salivary protein to persist within the arthropod and provide a paradigm for understanding how Rickettsia-like pathogens are maintained within vectors. PMID- 16717120 TI - Dopamine versus epinephrine for cardiovascular support in low birth weight infants: analysis of systemic effects and neonatal clinical outcomes. AB - BACKGROUND: Early postnatal adaptation to transitional circulation in low birth weight infants frequently is associated with low blood pressure and decreased blood flow to organs. Catecholamines have been used widely as treatment, despite remarkably little empirical evidence on the effects of vasopressor/inotropic support on circulation and on clinically important outcomes in sick newborn infants. AIMS: To explore the effectiveness of low/moderate-dose dopamine and epinephrine in the treatment of early systemic hypotension in low birth weight infants, evaluate the frequency of adverse drug effects, and examine neonatal clinical outcomes of patients in relation to treatment. DESIGN/METHODS: Newborns of <1501-g birth weight or <32 weeks of gestational age, with a mean blood pressure lower than gestational age in the first 24 hours of life, were assigned randomly to receive dopamine (2.5, 5, 7.5, and 10 microg/kg per minute; n = 28) or epinephrine (0.125, 0.250, 0.375, and 0.5 microg/kg per minute; n = 32) at doses that were increased stepwise every 20 minutes until optimal mean blood pressure was attained and maintained (responders). If this treatment was unsuccessful (nonresponders), sequential rescue therapy was started, consisting first of the addition of the second study drug and then hydrocortisone. OUTCOME MEASURES: These included: (1) short-term changes (first 96 hours, only responders) in heart rate, mean blood pressure, acid-base status, lactate, glycemia, urine output, and fluid-carbohydrate debit; and (2) medium-term morbidity, enteral nutrition tolerance, gastrointestinal complications, severity of lung disease, patent ductus arteriosus, cerebral ultrasound diagnoses, retinopathy of prematurity, and mortality. RESULTS: Patients enrolled in this trial did not differ in birth weight or gestational age (1008 +/- 286 g and 28.3 +/- 2.3 weeks in the dopamine group; 944 +/- 281 g and 27.7 +/- 2.4 weeks in the epinephrine group). Other main antenatal variables were also comparable. However, responders and nonresponders differed significantly with respect to the need for cardiorespiratory resuscitation at birth (3% vs 23%), Critical Risk Index for Babies score (3.8 +/- 3 vs 7 +/- 5), and premature rupture of membranes >24 hours (39.5% vs 13.6%), respectively. No differences were found in the rate of treatment failure (dopamine: 36%; epinephrine: 37%) or need for rescue therapy according to treatment allocation. Groups did not differ in age at initiation of therapy (dopamine: 5.3 +/- 3.9 hours; epinephrine: 5.2 +/- 3.3 hours), but withdrawal was significantly later in the dopamine group. For short-term changes, mean blood pressure showed a significant increase from baseline throughout the first 96 hours with no differences between groups. However, epinephrine produced a greater increase in heart rate than dopamine. After treatment began, epinephrine patients showed higher plasma lactate (first 36 hours) and lower bicarbonate and base excess (first 6 hours) and received more bicarbonate. Patients in the epinephrine group also had higher glycemia (first 24 hours) and needed insulin therapy more often. Groups did not differ in urine output or fluid carbohydrate supply during the first 96 hours. For medium-term morbidity, there were no differences in neonatal clinical outcomes in responders. However, significant differences were found in the incidence of patent ductus arteriosus, bronchopulmonary dysplasia, need for high-frequency ventilation, occurrence of necrotizing enterocolitis, and death between responders and nonresponders. CONCLUSIONS: Low/moderate-dose epinephrine is as effective as low/moderate-dose dopamine for the treatment of hypotension in low birth weight infants, although it is associated with more transitory adverse effects. PMID- 16717121 TI - Salmonella enterica serotype Choleraesuis infections in pediatric patients. AB - OBJECTIVE: Among >2500 nontyphoid Salmonella serotypes, Salmonella enterica serotype Choleraesuis shows a high predilection to cause systemic infections in humans. The objective of this study was to delineate the clinical and microbiologic features of pediatric patients with Salmonella Choleraesuis infections. METHODS: Between May 1999 and February 2003, a total of 33 patients who were <18 years of age had culture-confirmed S Choleraesuis infections. Clinical features, laboratory values, treatment, outcome, and antimicrobial susceptibility patterns of the bacterial isolates were analyzed. RESULTS: There were 24 males and 9 females with a mean age of 3 years. Fever (rectal temperature > or =38 degrees C; 94%) was the most common clinical presentation. Sixteen (52%) had fever lasting >5 days before admission. Only 18 (54%) patients had diarrhea. The most common mode of infection is occult bacteremia without focal infection. Compared with data obtained from adult patients, the gastrointestinal manifestations appeared more frequently seen in pediatric patients. However, among the 18 who presented with diarrhea, 14 had concomitant bloodstream infection. Only 1 patient, who was a case of acute leukemia, died of S Choleraesuis sepsis. Resistance to ceftriaxone, ciprofloxacin, ampicillin, trimethoprim-sulfamethoxazole, and chloramphenicol was found in 6%, 28%, 88%, 76%, and 83% of the isolates, respectively. CONCLUSION: Children with S Choleraesuis infections usually presented with occult bacteremia with mild gastrointestinal involvement. The mortality of S Choleraesuis infections in previously healthy children is low. Ciprofloxacin resistance among S Choleraesuis isolates from pediatric patients was lower than that of isolates from adult patients. In view of the high rate of multidrug resistance, third-generation cephalosporins seem to be the drug of choice for treatment of invasive S Choleraesuis infections. PMID- 16717122 TI - Cardiac involvement in Fukuyama-type congenital muscular dystrophy. AB - BACKGROUND: Fukuyama-type congenital muscular dystrophy is an autosomal recessive disorder characterized by generalized skeletal muscle weakness and hypotonia from early infancy and by mental retardation. Little is known about cardiac involvement in patients with Fukuyama-type congenital muscular dystrophy. This study evaluated whether cardiac involvement exists in patients with Fukuyama-type congenital muscular dystrophy. METHODS AND RESULTS: We evaluated left ventricular function using M-mode and Doppler echocardiography in 34 patients with Fukuyama type congenital muscular dystrophy. The age ranged from 6 months to 30 years (median: 6 years). A total of 64 recordings were analyzed. Left ventricular dimensions and parameters of systolic function measured included left ventricular end-diastolic dimension, left ventricular fractional shortening, left ventricular wall thickness, and the mean velocity of circumferential fiber shortening and end systolic wall stress relationship. Left ventricular end-diastolic dimension z score >2 was observed in 2 patients (6%). Left ventricular fractional shortening <0.28 and/or reduced mean velocity of circumferential fiber shortening in the mean velocity of circumferential fiber shortening-end-systolic wall stress relationship were observed in 16 patients (47%). A significant correlation between age and left ventricular fractional shortening was observed, and left ventricular fractional shortening decreased with age. Of 12 patients >15 years of age, 10 (83%) showed decreased left ventricular systolic function. Left ventricular fractional shortening was normal in most patients <10 years of age, and it was reduced in most patients >15 years of age. Five patients died of heart failure or respiratory problems, and a histologic examination confirmed the presence of myocardial fibrosis. No patients showed increased left ventricular wall thickness or a conduction abnormality on electrocardiograms. CONCLUSION: Cardiac involvement exists in patients with Fukuyama-type congenital muscular dystrophy and becomes evident in older children in the second decade. A cardiac evaluation, including echocardiograms and subsequent follow-up, is important, especially in patients >10 years of age. PMID- 16717123 TI - Paradigm lost: milton connects kinesin heavy chain to miro on mitochondria. AB - The kinesin motor typically binds to cargo through its light chains. In this issue Glater et al. demonstrate a new type of linkage through the adapter protein, milton, and the mitochondrial membrane GTPase, miro. This is an important result because it represents a new mechanism of cargo binding and because miro's ability to bind GTP and calcium suggests that it is involved in the regulation of mitochondrial transport. PMID- 16717124 TI - Direct membrane protein-DNA interactions required early in nuclear envelope assembly. AB - Among the earliest events in postmitotic nuclear envelope (NE) assembly are the interactions between chromatin and the membranes that will fuse to form the NE. It has been proposed that interactions between integral NE proteins and chromatin proteins mediate initial membrane recruitment to chromatin. We show that several transmembrane NE proteins bind to DNA directly and that NE membrane proteins as a class are enriched in long, basic domains that potentially bind DNA. Membrane fractions that are essential for NE formation are shown to bind directly to protein-free DNA, and our data suggest that these interactions are critical for early steps in NE assembly. PMID- 16717125 TI - Structural damage to meiotic chromosomes impairs DNA recombination and checkpoint control in mammalian oocytes. AB - Meiosis in human oocytes is a highly error-prone process with profound effects on germ cell and embryo development. The synaptonemal complex protein 3 (SYCP3) transiently supports the structural organization of the meiotic chromosome axis. Offspring derived from murine Sycp3(-)(/)(-) females die in utero as a result of aneuploidy. We studied the nature of the proximal chromosomal defects that give rise to aneuploidy in Sycp3(-)(/)(-) oocytes and how these errors evade meiotic quality control mechanisms. We show that DNA double-stranded breaks are inefficiently repaired in Sycp3(-)(/)(-) oocytes, thereby generating a temporal spectrum of recombination errors. This is indicated by a strong residual gammaH2AX labeling retained at late meiotic stages in mutant oocytes and an increased persistence of recombination-related proteins associated with meiotic chromosomes. Although a majority of the mutant oocytes are rapidly eliminated at early postnatal development, a subset with a small number of unfinished crossovers evades the DNA damage checkpoint, resulting in the formation of aneuploid gametes. PMID- 16717126 TI - Mouse SYCP2 is required for synaptonemal complex assembly and chromosomal synapsis during male meiosis. AB - During meiosis, the arrangement of homologous chromosomes is tightly regulated by the synaptonemal complex (SC). Each SC consists of two axial/lateral elements (AEs/LEs), and numerous transverse filaments. SC protein 2 (SYCP2) and SYCP3 are integral components of AEs/LEs in mammals. We find that SYCP2 forms heterodimers with SYCP3 both in vitro and in vivo. An evolutionarily conserved coiled coil domain in SYCP2 is required for binding to SYCP3. We generated a mutant Sycp2 allele in mice that lacks the coiled coil domain. The fertility of homozygous Sycp2 mutant mice is sexually dimorphic; males are sterile because of a block in meiosis, whereas females are subfertile with sharply reduced litter size. Sycp2 mutant spermatocytes exhibit failure in the formation of AEs and chromosomal synapsis. Strikingly, the mutant SYCP2 protein localizes to axial chromosomal cores in both spermatocytes and fetal oocytes, but SYCP3 does not, demonstrating that SYCP2 is a primary determinant of AEs/LEs and, thus, is required for the incorporation of SYCP3 into SCs. PMID- 16717127 TI - The origin and maintenance of mammalian peroxisomes involves a de novo PEX16 dependent pathway from the ER. AB - Peroxisomes are ubiquitous organelles that proliferate under different physiological conditions and can form de novo in cells that lack them. The endoplasmic reticulum (ER) has been shown to be the source of peroxisomes in yeast and plant cells. It remains unclear, however, whether the ER has a similar role in mammalian cells and whether peroxisome division or outgrowth from the ER maintains peroxisomes in growing cells. We use a new in cellula pulse-chase imaging protocol with photoactivatable GFP to investigate the mechanism underlying the biogenesis of mammalian peroxisomes. We provide direct evidence that peroxisomes can arise de novo from the ER in both normal and peroxisome-less mutant cells. We further show that PEX16 regulates this process by being cotranslationally inserted into the ER and serving to recruit other peroxisomal membrane proteins to membranes. Finally, we demonstrate that the increase in peroxisome number in growing wild-type cells results primarily from new peroxisomes derived from the ER rather than by division of preexisting peroxisomes. PMID- 16717128 TI - Distinct p53 acetylation cassettes differentially influence gene-expression patterns and cell fate. AB - The activity of the p53 gene product is regulated by a plethora of posttranslational modifications. An open question is whether such posttranslational changes act redundantly or dependently upon one another. We show that a functional interference between specific acetylated and phosphorylated residues of p53 influences cell fate. Acetylation of lysine 320 (K320) prevents phosphorylation of crucial serines in the NH(2)-terminal region of p53; only allows activation of genes containing high-affinity p53 binding sites, such as p21/WAF; and promotes cell survival after DNA damage. In contrast, acetylation of K373 leads to hyperphosphorylation of p53 NH(2)-terminal residues and enhances the interaction with promoters for which p53 possesses low DNA binding affinity, such as those contained in proapoptotic genes, leading to cell death. Further, acetylation of each of these two lysine clusters differentially regulates the interaction of p53 with coactivators and corepressors and produces distinct gene-expression profiles. By analogy with the "histone code" hypothesis, we propose that the multiple biological activities of p53 are orchestrated and deciphered by different "p53 cassettes," each containing combination patterns of posttranslational modifications and protein-protein interactions. PMID- 16717129 TI - Axonal transport of mitochondria requires milton to recruit kinesin heavy chain and is light chain independent. AB - Mitochondria are distributed within cells to match local energy demands. We report that the microtubule-dependent transport of mitochondria depends on the ability of milton to act as an adaptor protein that can recruit the heavy chain of conventional kinesin-1 (kinesin heavy chain [KHC]) to mitochondria. Biochemical and genetic evidence demonstrate that kinesin recruitment and mitochondrial transport are independent of kinesin light chain (KLC); KLC antagonizes milton's association with KHC and is absent from milton-KHC complexes, and mitochondria are present in klc (-/-) photoreceptor axons. The recruitment of KHC to mitochondria is, in part, determined by the NH(2) terminus splicing variant of milton. A direct interaction occurs between milton and miro, which is a mitochondrial Rho-like GTPase, and this interaction can influence the recruitment of milton to mitochondria. Thus, milton and miro are likely to form an essential protein complex that links KHC to mitochondria for light chain independent, anterograde transport of mitochondria. PMID- 16717130 TI - Paxillin phosphorylation at Ser273 localizes a GIT1-PIX-PAK complex and regulates adhesion and protrusion dynamics. AB - Continuous adhesion formation and disassembly (adhesion turnover) in the protrusions of migrating cells is regulated by unclear mechanisms. We show that p21-activated kinase (PAK)-induced phosphorylation of serine 273 in paxillin is a critical regulator of this turnover. Paxillin-S273 phosphorylation dramatically increases migration, protrusion, and adhesion turnover by increasing paxillin GIT1 binding and promoting the localization of a GIT1-PIX-PAK signaling module near the leading edge. Mutants that interfere with the formation of this ternary module abrogate the effects of paxillin-S273 phosphorylation. PAK-dependent paxillin-S273 phosphorylation functions in a positive-feedback loop, as active PAK, active Rac, and myosin II activity are all downstream effectors of this turnover pathway. Finally, our studies led us to identify in highly motile cells a class of small adhesions that reside near the leading edge, turnover in 20-30 s, and resemble those seen with paxillin-S273 phosphorylation. These adhesions appear to be regulated by the GIT1-PIX-PAK module near the leading edge. PMID- 16717131 TI - Neisseria meningitidis infection of human endothelial cells interferes with leukocyte transmigration by preventing the formation of endothelial docking structures. AB - Neisseria meningitidis elicits the formation of membrane protrusions on vascular endothelial cells, enabling its internalization and transcytosis. We provide evidence that this process interferes with the transendothelial migration of leukocytes. Bacteria adhering to endothelial cells actively recruit ezrin, moesin, and ezrin binding adhesion molecules. These molecules no longer accumulate at sites of leukocyte-endothelial contact, preventing the formation of the endothelial docking structures required for proper leukocyte diapedesis. Overexpression of exogenous ezrin or moesin is sufficient to rescue the formation of docking structures on and leukocyte migration through infected endothelial monolayers. Inversely, expression of the dominant-negative NH(2)-terminal domain of ezrin markedly inhibits the formation of docking structures and leukocyte diapedesis through noninfected monolayers. Ezrin and moesin thus appear as pivotal endothelial proteins required for leukocyte diapedesis that are titrated away by N. meningitidis. These results highlight a novel strategy developed by a bacterial pathogen to hamper the host inflammatory response by interfering with leukocyte-endothelial cell interaction. PMID- 16717132 TI - Maintenance of HCV-specific T-cell responses in antibody-deficient patients a decade after early therapy. PMID- 16717133 TI - Complete regression of cutaneous lesions of refractory Ph+ ALL after 4 weeks of treatment with BMS-354825. PMID- 16717134 TI - Discordant distribution of JAK2V617F mutation in siblings with familial myeloproliferative disorders. PMID- 16717135 TI - Discovery of a novel shp2 protein tyrosine phosphatase inhibitor. AB - Shp2 is a nonreceptor protein tyrosine phosphatase (PTP) encoded by the PTPN11 gene. It is involved in growth factorinduced activation of mitogen-activated protein (MAP) kinases Erk1 and Erk2 (Erk1/2) and has been implicated in the pathogenicity of the oncogenic bacterium Helicobacter pylori. Moreover, gain-of function Shp2 mutations have been found in childhood leukemias and Noonan syndrome. Thus, small molecule Shp2 PTP inhibitors are much needed reagents for evaluation of Shp2 as a therapeutic target and for chemical biology studies of Shp2 function. By screening the National Cancer Institute (NCI) Diversity Set chemical library, we identified 8-hydroxy-7-(6-sulfonaphthalen-2-yl)diazenyl quinoline-5-sulfonic acid (NSC-87877) as a potent Shp2 PTP inhibitor. Molecular modeling and site-directed mutagenesis studies suggested that NSC-87877 binds to the catalytic cleft of Shp2 PTP. NSC-87877 cross-inhibited Shp1 in vitro, but it was selective for Shp2 over other PTPs (PTP1B, HePTP, DEP1, CD45, and LAR). It is noteworthy that NSC-87877 inhibited epidermal growth factor (EGF)-induced activation of Shp2 PTP, Ras, and Erk1/2 in cell cultures but did not block EGF induced Gab1 tyrosine phosphorylation or Gab1-Shp2 association. Furthermore, NSC 87877 inhibited Erk1/2 activation by a Gab1-Shp2 chimera but did not affect the Shp2-independent Erk1/2 activation by phorbol 12-myristate 13-acetate. These results identified NSC-87877 as the first PTP inhibitor capable of inhibiting Shp2 PTP in cell cultures without a detectable off-target effect. Our study also provides the first pharmacological evidence that Shp2 mediates EGF-induced Erk1/2 MAP kinase activation. PMID- 16717136 TI - Divergent activities of human glutathione transferases in the bioactivation of azathioprine. AB - Azathioprine is a thiopurine prodrug clinically used for immunosuppression in the treatment of inflammatory diseases and in pharmacological regimens of organ transplantations. Its pharmacological action is based on the release of 6 mercaptopurine, but the biochemical processes underlying this biotransformation have remained obscure. In this investigation, human glutathione transferases (GSTs) from seven distinct classes were assayed with azathioprine. GSTs A1-1, A2 2, and M1-1, all abundantly expressed in human liver, displayed the highest activity among the 14 GSTs tested. The uncatalyzed reaction of azathioprine with glutathione was estimated to be less than 1% of the GST-catalyzed biotransformation. GST M1-1 is polymorphic with a frequently occurring null allele, and GSTs A1-1 and A2-2 show variable expression levels in human subjects, implying significant differences in the rate of 6-mercaptopurine release from azathioprine. Individuals expressing high GST activity are apparently predisposed for adverse reactions to azathioprine treatment, both by promoting excessively high concentrations of free 6-mercaptopurine and its toxic metabolites and by depleting cellular glutathione. These novel aspects of GST-dependent azathioprine biotransformation have not been considered previously. PMID- 16717137 TI - Brief report: the impact of maternal posttraumatic stress disorder symptoms and child gender on risk for persistent posttraumatic stress disorder symptoms in child trauma victims. AB - OBJECTIVE: To longitudinally examine the impact of maternal posttraumatic stress disorder symptoms (PTSS) on child adjustment following a child's traumatic injury, focusing on child gender differences. METHODS: Forty-one child traumatic injury victims aged 8-18 years and their biological mothers were interviewed over two follow-ups (6 weeks and 7 months). Children were administered the Clinician Administered Posttraumatic Stress Disorder (PTSD) Scale for Children and Adolescents (CAPS-CA), whereas mothers completed the CAPS. RESULTS: Six weeks post trauma, maternal PTSS were significantly related to PTSS in boys but not in girls. However, at 7 months, maternal PTSS were strongly related to child PTSS in both boys and girls. Significant 6-week maternal distress-child gender interactions suggested that maternal PTSS, especially avoidance, predicted greater 7-month PTSS but that this was primarily because of a significant relationship in females. CONCLUSIONS: Maternal distress was found to negatively impact subsequent child adjustment, particularly in females. These results underscore the importance of considering family-centered interventions for child PTSD, especially in girls. PMID- 16717138 TI - Appraisal and coping with daily stressors by pediatric patients with chronic abdominal pain. AB - OBJECTIVE: To test the hypothesis that pain patients differ from well children in their appraisal and coping with daily stressors and to test a model of the relation of stress appraisal and coping to symptoms and disability. METHODS: Pediatric patients with chronic abdominal pain (n = 143) and well children (n = 104) completed a 5-day diary study regarding their appraisal and coping with daily stressors. Somatic symptoms, depressive symptoms, and functional disability were assessed 2 months later. RESULTS: Compared to well children, pain patients were less confident of their ability either to change or to adapt to stress and were less likely to use accommodative coping strategies. Different patterns of stress appraisal were associated with active, passive, and accommodative coping. Both appraisals and coping were significantly related to symptoms and disability. CONCLUSIONS: The relation between stress and symptoms in pediatric pain patients may be explained in part by their appraisal and coping with stressors. The relation between appraisal and coping was consistent with Lazarus, R. S., & Folkman, S. (1984). Stress, Appraisal, and Coping. New York: Springer. PMID- 16717139 TI - Decision-making competence and adherence to treatment in adolescents with diabetes. AB - OBJECTIVE: To examine decision-making competence (the ability to form effective plans for managing different situations) in a sample of adolescents with type I diabetes and their parents. We hypothesized that adolescent decision-making competence would mediate the relationship between parent-adolescent communication and adherence to treatment. METHODS: The sample consisted of 63 adolescents and their parents. Parent-adolescent communication during a problem-solving task was assessed, as well as adolescent maladaptive decision-making (adolescent report), adherence to treatment (parent and provider report; number of glucose tests), and metabolic control (HbA1C). RESULTS: Parent-adolescent communication was associated with adherence to treatment but not with adolescent decision-making. Poorer decision-making was associated with lower adherence per parent report but not provider report or the number of glucose tests. Decision-making competence did not mediate the relationships between parent-adolescent communication and adherence. CONCLUSIONS: These results are consistent with prior research demonstrating associations between parent-adolescent communication and adherence and identify adolescent decision-making competence as another potentially important correlate of adherence. These findings highlight several areas for future research. PMID- 16717140 TI - Pediatric asthma morbidity: the importance of symptom perception and family response to symptoms. AB - OBJECTIVE: To determine whether family response to asthma symptoms mediates the relationship between child symptom perception and morbidity. METHODS: A total of 122 children with asthma, aged between 7 and 17 years (47% females; 25% ethnic minorities), were recruited from three sites. Participants completed a family asthma management interview and 5-6 weeks of symptom perception assessment. RESULTS: Family response to symptoms mediated the relationship between child underestimation of symptoms and asthma morbidity and partially mediated the relationship between accurate symptom perception and morbidity. In contrast, although child overestimation of symptoms and family response to symptoms were independently related to asthma morbidity, a mediational model was not supported. CONCLUSIONS: Our study found support for the role of family response to symptoms in mediating the relationship between child symptom perception and morbidity, particularly with regard to underestimation of symptoms, underscoring the need for behavioral tools to accurately recognize and optimally respond to exacerbations. PMID- 16717143 TI - Brachial and central arterial pressure. PMID- 16717141 TI - Functional noncoding sequences derived from SINEs in the mammalian genome. AB - Recent comparative analyses of mammalian sequences have revealed that a large number of nonprotein-coding genomic regions are under strong selective constraint. Here, we report that some of these loci have been derived from a newly defined family of ancient SINEs (short interspersed repetitive elements). This is a surprising result, as SINEs and other transposable elements are commonly thought to be genomic parasites. We named the ancient SINE family AmnSINE1, for Amniota SINE1, because we found it to be present in mammals as well as in birds, and some copies predate the mammalian-bird split 310 million years ago (Mya). AmnSINE1 has a chimeric structure of a 5S rRNA and a tRNA-derived SINE, and is related to five tRNA-derived SINE families that we characterized here in the coelacanth, dogfish shark, hagfish, and amphioxus genomes. All of the newly described SINE families have a common central domain that is also shared by zebrafish SINE3, and we collectively name them the DeuSINE (Deuterostomia SINE) superfamily. Notably, of the approximately 1000 still identifiable copies of AmnSINE1 in the human genome, 105 correspond to loci phylogenetically highly conserved among mammalian orthologs. The conservation is strongest over the central domain. Thus, AmnSINE1 appears to be the best example of a transposable element of which a significant fraction of the copies have acquired genomic functionality. PMID- 16717144 TI - Influences of norepinephrine transporter function on the distribution of sympathetic activity in humans. AB - Previous studies suggest that neuronal norepinephrine transporter function may regulate the distribution of sympathetic activity among blood vessels, heart, and kidney; we tested the functional relevance in humans. Sixteen healthy men (26+/-1 years) ingested 8 mg of the selective norepinephrine reuptake transporter inhibitor reboxetine or a matching placebo on 2 separate days in a double-blind, randomized, crossover fashion. We monitored heart rate, thoracic bioimpedance, blood pressure, glomerular filtration rate, and renal blood flow. Ninety minutes after ingestion of the test medication, subjects were tilted to a 45 degrees head up position, where they remained for an additional 30 minutes. Reboxetine increased supine systolic blood pressure through an increase in cardiac output whereas systemic vascular resistance decreased. Furthermore, reboxetine increased heart rate, particularly with a head-up tilt. Supine plasma renin activity was 0.71+/-0.15 ng angiotensin (Ang)/L per mL/h with placebo and 0.36+/-0.07 ngAng/L per mL/h with reboxetine (P<0.01). Supine plasma Ang II concentrations were also decreased with reboxetine. Both plasma renin activity and Ang II concentrations remained suppressed during head-up tilt. On placebo, renal vascular resistance increased with head-up tilt. The response was abolished with norepinephrine reuptake inhibition. We conclude that norepinephrine reuptake function profoundly influences the distribution of sympathetic activity between the heart, vasculature, and kidney in humans. All of these changes are physiologically relevant because they lead to corresponding changes in organ function. PMID- 16717145 TI - Telmisartan prevents obesity and increases the expression of uncoupling protein 1 in diet-induced obese mice. AB - The aim of the present study was to clarify the effect of telmisartan, an angiotensin II receptor blocker, on the development of obesity and related metabolic disorders in diet-induced obese mice. Treatment with telmisartan dissolved in drinking water at a dosage of 5 mg/kg per day for 14 days attenuated the diet-induced weight gain without affecting food intake in diet-induced obese mice compared with controls using nontreated water. Telmisartan treatment decreased the weight of visceral adipose tissue and the triglyceride content in the liver and skeletal muscle. In addition, hyperglycemia, hyperinsulinemia, and hypertriglyceridemia in diet-induced obese mice all improved with telmisartan treatment. Furthermore, telmisartan treatment increased adiponectin mRNA in visceral white adipose tissue and was associated with a concomitant change in the serum adiponectin level. In contrast, the treatment reduced the serum level of resistin. Finally, telmisartan treatment increased the mRNA expression of uncoupling protein 1 in brown adipose tissue and was accompanied by an increase in oxygen consumption. In conclusion, telmisartan treatment might prevent the development of obesity and related metabolic disorders by altering the levels of adiponectin, resistin, and uncoupling protein 1 in diet-induced obese mice. Our results indicate that telmisartan can be used as a therapeutic tool for metabolic syndrome, including visceral obesity. PMID- 16717146 TI - 11beta-hydroxysteroid dehydrogenase type 2 activity in hypothalamic paraventricular nucleus modulates sympathetic excitation. AB - Aldosterone stimulates the sympathetic nervous system by binding to a select population of brain mineralocorticoid receptors (MR). These MR have an equal affinity for corticosterone that is present in substantially higher concentrations, but are held in reserve for aldosterone by activity of the enzyme 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD-2), which converts corticosterone to an inactive metabolite. Thus, colocalization of MR and 11beta HSD-2 activity may help identify brain regions that mediate the effects of aldosterone. The present studies tested the hypothesis that 11beta-HSD-2 activity regulates MR-mediated responses in the paraventricular nucleus (PVN) of the hypothalamus, a forebrain region implicated in sympathetic regulation. Real-time polymerase chain reaction revealed the presence of 11beta-HSD-2 mRNA in PVN. In anesthetized adult male Sprague-Dawley rats, microinjection of the 11beta-HSD-2 inhibitor carbenoxolone (CBX) into PVN increased mean arterial pressure, heart rate, and renal sympathetic nerve activity. Intracerebroventricular injections of CBX excited PVN neurons and increased mean arterial pressure, heart rate, and renal sympathetic nerve activity. The ability of CBX to increase sympathetic activity by inhibiting 11beta-HSD-2, thereby permitting corticosterone to activate MR, was confirmed by the following: Intracerebroventricular glycyrrhizic acid, another 11beta-HSD-2 inhibitor, mimicked the sympathoexcitatory effects of CBX; the sympathoexcitatory effects of CBX were blocked by spironolactone, a MR antagonist. Neither CBX nor glycyrrhizic acid elicited a response in adrenalectomized rats. These findings suggest that MR in PVN contribute to sympathetic regulation and may be activated by aldosterone or corticosterone (or cortisol in humans) depending on the state of 11beta-HSD-2 activity. PMID- 16717147 TI - Effects of nitroglycerin on erythrocyte rheology and oxygen unloading: novel role of S-nitrosohemoglobin in relieving myocardial ischemia. AB - BACKGROUND: We hypothesized that nitroglycerin improves O2 delivery to ischemic tissue by altering erythrocyte rheology and O2 unloading through an increase in bioactive nitric oxide (NO) content. METHODS AND RESULTS: Twelve dogs with resting flow-reducing single-vessel stenosis were studied at rest and during intracoronary infusion of nitroglycerin (0.3 to 0.6 microg.kg(-1).min(-1)). Half the dogs also had occlusion of the remote coronary artery to remove any collateral effects. Systemic and coronary hemodynamics, myocardial blood flow (MBF), whole blood viscosity (WBeta), erythrocyte charge (EC) and mobility (EM), regional myocardial O2 delivery and consumption, and tissue O2 pressure (Po2) were measured. No changes in systemic hemodynamics were seen with nitroglycerin. Despite flow-limiting stenosis, MBF increased significantly in the central 25% of the ischemic bed, which was associated with an approximately 19% decrease in WBeta. There was a good correlation (r=0.87) between the two. The decrease in WBeta was associated with a decrease in EC and an increase in EM (r=0.83). The nitroglycerin-induced increase in tissue Po2 was disproportionate to the increase in MBF, indicating enhanced O2 unloading. Erythrocyte S-nitrosothiol content (reflecting mainly S-nitrosohemoglobin) was significantly higher for blood exposed in vitro to 0.1 micromol/L nitroglycerin or the NO donor SNAP, as compared with control (18.9+/-8.8 and 10.5+/-6.5 versus 2.6+/-0.5x10(-5), P<0.05). CONCLUSIONS: The augmented MBF in the ischemic microcirculation during nitroglycerin administration occurs in tandem with increased erythrocyte S nitrosothiol content, EM, and O2 unloading. These additional microvascular mechanisms may contribute to the powerful antiischemic effects of nitroglycerin, especially during low-flow states. PMID- 16717148 TI - Relationship of BMPR2 mutations to vasoreactivity in pulmonary arterial hypertension. AB - BACKGROUND: Vasoreactivity tests are fundamental in evaluating pulmonary arterial hypertension (PAH). Mutations of the transforming growth factor-beta type II receptor gene, BMPR2, predispose to the development of pulmonary hypertension and may alter the response to vasodilators. Previous investigations have not examined the relationship of BMPR2 mutations to vasoreactivity. METHODS AND RESULTS: We identified 133 consecutive unrelated patients with either idiopathic or familial PAH. Sixty-six patients were excluded because we lacked either DNA samples (n=18) or complete data from a vasoreactivity test (n=48). The remaining 67 patients were screened for BMPR2 DNA sequence variations, and specific variations were confirmed by gene sequencing. The vasoreactivity of patients with nonsynonymous BMPR2 variations was compared with that of patients without nonsynonymous BMPR2 variations. We found nonsynonymous BMPR2 variations in 27 of 67 patients with idiopathic (n=16 of 52) or familial (n=11 of 15) PAH. Vasoreactivity was identified in 3.7% of 27 patients with nonsynonymous BMPR2 variations and in 35% of 40 patients without nonsynonymous BMPR2 variations (P=0.003). Five of the 27 nonsynonymous variations occur commonly in healthy individuals. None of the remaining 22 patients with BMPR2 variations demonstrated vasoreactivity, and the analysis remained unchanged when we assumed that nonsynonymous BMPR2 variations were present in all 15 patients with familial PAH. CONCLUSIONS: Patients with familial or idiopathic PAH and nonsynonymous BMPR2 variations are unlikely to demonstrate vasoreactivity. Further trials are required to determine whether long term therapy can be directed by tests for BMPR2 variations. PMID- 16717149 TI - Enhanced ventricular untwisting during exercise: a mechanistic manifestation of elastic recoil described by Doppler tissue imaging. AB - BACKGROUND: The cascade of events by which early diastolic left ventricular (LV) filling increases with exercise is not fully elucidated. Doppler tissue imaging (DTI) can detect myocardial motion, including torsion, whereas color M-mode Doppler (CMM) can quantify LV intraventricular pressure gradients (IVPGs). METHODS AND RESULTS: Twenty healthy volunteers underwent echocardiographic examination with DTI at rest and during submaximal supine bicycle exercise. We assessed LV long-/short-axis function, torsion, volume, inflow dynamics, and early diastolic IVPG derived from CMM data. LV torsion and untwisting velocity increased with exercise (torsion, 11+/-4 degrees to 24+/-8 degrees ; untwisting velocity, -2.0+/-0.7 to -5.6+/-2.3 rad/s) that was associated with an increase in IVPG (1.4+/-0.5 to 3.7+/-1.2 mm Hg). Untwisting in normal subjects occurred during isovolumic relaxation and early filling, significantly before long-axis lengthening or radial expansion. The clinical feasibility of this method was tested in 7 patients with hypertrophic cardiomyopathy (HCM); torsion was higher at rest but did not increase with exercise (16+/-4 degrees to 14+/-6 degrees), whereas untwisting was delayed and unenhanced (-1.6+/-0.8 to -2.3+/-1.2 rad/s). In concert, IVPG was similar at rest (1.2+/-0.3 mm Hg), but the exercise response was blunted (1.6+/-0.8 mm Hg). In normal subjects and HCM patients, there was a similar linear relation between IVPG and untwisting rate, with an overall correlation coefficient of r=0.75 (P<0.0001). CONCLUSIONS: LV untwisting appears to be linked temporally with early diastolic base-to-apex pressure gradients, enhanced by exercise, which may assist efficient LV filling, an effect that appears blunted in HCM. Thus, LV torsion and subsequent rapid untwisting appear to be manifestations of elastic recoil, critically linking systolic contraction to diastolic filling. PMID- 16717150 TI - Significance of a fragmented QRS complex versus a Q wave in patients with coronary artery disease. AB - BACKGROUND: Q waves on a 12-lead ECG are markers of a prior myocardial infarction (MI). However, they may regress or even disappear over time, and there is no specific ECG sign of a non-Q-wave MI. Fragmented QRS complexes (fQRSs), which include various RSR' patterns, without a typical bundle-branch block are markers of altered ventricular depolarization owing to a prior myocardial scar. We postulated that the presence of an fQRS might improve the ability to detect a prior MI compared with Q waves alone by ECG. METHODS AND RESULTS: A cohort of 479 consecutive patients (mean+/-SD age, 58.2+/-13.2 years; 283 males) who were referred for nuclear stress tests was studied. The fQRS included various morphologies of the QRS (<120 ms), which included an additional R wave (R') or notching in the nadir of the S wave, or >1 R' (fragmentation) in 2 contiguous leads, corresponding to a major coronary artery territory. The Q wave was present in 71 (14.8%) patients, an fQRS was present in 191 (34.9%) patients, and an fQRS and/or a Q wave was present in 203 (42.3%) patients. Sensitivity, specificity, and the negative predictive value for myocardial scar as detected by single photon emission computed tomography analysis were 36.3%, 99.2%, and 70.8%, respectively, for the Q wave alone; 85.6%, 89%, and 92.7%, respectively, for the fQRS; and 91.4%, 89%, and 94.2%, respectively, for the Q wave and/or fQRS. CONCLUSIONS: The fQRS on a 12-lead ECG is a marker of a prior MI, defined by regional perfusion abnormalities, which has a substantially higher sensitivity and negative predictive value compared with the Q wave. PMID- 16717151 TI - Percutaneous treatment with drug-eluting stent implantation versus bypass surgery for unprotected left main stenosis: a single-center experience. AB - BACKGROUND: Improvements in results with percutaneous coronary intervention (PCI) with drug-eluting stents (DES) may extend their use in patients with left main coronary artery (LMCA) stenosis. METHODS AND RESULTS: Two hundred forty-nine patients with LMCA stenosis were treated with PCI and DES implantation (n=107) or coronary artery bypass grafting (CABG) (n=142), in a single center, between March 2002 and July 2004. A propensity analysis was performed to adjust for baseline differences between the two cohorts. At 1 year, there was no statistical difference in the occurrence of death in PCI versus CABG both for the unadjusted (OR=0.291; 95% CI=0.054 to 1.085; P=0.0710) and adjusted analyses (OR=0.331; 95% CI=0.055 to 1.404; P=0.1673). PCI was correlated to a lower occurrence of the composite end points of death and myocardial infarction (unadjusted OR=0.235; 95% CI=0.048 to 0.580; P=0.0002; adjusted OR=0.260; 95% CI=0.078 to 0.597; P=0.0005) and death, myocardial infarction, and cerebrovascular events (unadjusted OR=0.300; 95% CI=0.102 to 0.617; P=0.0004; adjusted OR=0.385; 95% CI=0.180 to 0.819; P=0.01). No difference was detected in the occurrence of major adverse cardiac and cerebrovascular event at the unadjusted (OR=0.675; 95% CI=0.371 to 1.189; P=0.1891) and adjusted analyses (OR=0.568; 95% CI=0.229 to 1.344; P=0.2266). CONCLUSIONS: At 1 year, in this single-center, retrospective experience, there was no difference in the degree of protection against death, stroke, myocardial infarction, and revascularization between PCI with DES and CABG for LMCA disease. PMID- 16717152 TI - Impact of transforming growth factor-beta1 on atrioventricular node conduction modification by injected autologous fibroblasts in the canine heart. AB - BACKGROUND: Atrioventricular (AV) nodal ablation for management of atrial fibrillation (AF) is irreversible and requires permanent pacemaker implantation. We hypothesized that as an alternative, implantation of autologous fibroblasts in the perinodal region would focally modify AV nodal conduction and that this modulation would be enhanced by pretreatment with transforming growth factor beta1 (TGF-beta1), a stimulant of fibroblasts. METHODS AND RESULTS: Skin biopsies were taken from 12 mongrel dogs, and derived fibroblasts were dissociated and grown in culture for 2 weeks. Multiple injections (0.25 mL) were made through an 8F NOGA catheter along the fast/slow AV nodal pathways as guided by an electroanatomic mapping system. Seven dogs received fibroblasts alone (1x10(6) cells/mL), 7 dogs received TGF-beta1 (5 microg), 4 dogs received fibroblasts and TGF-beta1 (1x10(6) cells/mL+5 microg), and 4 dogs received saline only. AV node function was assessed at baseline and after 4 weeks. Saline (80 mL) with assigned therapy (0.25 mL per injection) was injected into the peri-AV nodal region in each dog. At baseline, the AH interval (66+/-3 ms) and the average RR interval (331+/-17 ms) in pacing-induced AF were similar in each cohort. The increase in AH interval in normal sinus rhythm was longer after fibroblast (23+/-4 versus 5+/ 5 ms; P=0.05) and fibroblast plus TGF-beta1 (50+/-5 versus 5+/-5 ms; P<0.001) injections than with saline alone, with similar findings during high right atrium and distal coronary sinus pacing. The AH interval was not significantly increased after TGF-beta1 injections. The AH interval was significantly longer after fibroblast plus TGF-beta1 injections than with either therapy (TGF-beta1 or fibroblasts) alone. The RR interval during AF was increased in dogs that received fibroblasts alone (110+/-36 versus -41+/-34 ms) and to a greater extent with the addition of TGF-beta1 (294+/-108 versus -41+/-34 ms). No AV block was seen in any cohort at 4 weeks. Labeled fibroblasts that expressed vimentin were identified in all dogs that received cell injections at 4 weeks. CONCLUSIONS: AV nodal modification can be achieved with injected fibroblasts without the creation of AV block. The effect on AV node conduction is substantially enhanced by pretreatment of fibroblasts with TGF-beta1. These data have therapeutic potential for the management of rapid ventricular rate during AF without pacemaker implantation. PMID- 16717153 TI - Inhibiting p90 ribosomal S6 kinase prevents (Na+)-H+ exchanger-mediated cardiac ischemia-reperfusion injury. AB - BACKGROUND: Pharmacological and genetic studies indicate that the (Na+)-H+ exchanger isoform 1 (NHE1) plays a critical role in myocardial ischemia and reperfusion (I/R) injury. We found that p90 ribosomal S6 kinase (RSK) phosphorylated serine 703 of NHE1, stimulating 14-3-3 binding and NHE1 activity. Therefore, we hypothesized that inhibiting RSK in cardiomyocytes would prevent NHE1 activation and decrease I/R-mediated injury. METHODS AND RESULTS: To examine the role of RSK in vivo, we generated transgenic mice with cardiac-specific overexpression of dominant negative RSK (DN-RSK-TG). DN-RSK-TG hearts demonstrated normal basal cardiac function and morphology. However, myocardial infarction (left coronary artery occlusion for 45 minutes) in DN-RSK-TG hearts was significantly reduced at 24 hours of reperfusion from 46.9+/-5.6% area at risk in nontransgenic littermate controls to 26.0+/-4.2% in DN-RSK-TG (P<0.01). Cardiomyocyte apoptosis was significantly reduced after I/R in DN-RSK (0.9+/ 0.2%) compared with nontransgenic littermate controls (6.2+/-2.6%). Importantly, activation of RSK and interaction of 14-3-3 with NHE1, necessary for agonist stimulated NHE1 activity, were increased by I/R and inhibited by 70% in DN-RSK-TG (P<0.01). Next, we transduced rat neonatal cardiomyocytes with adenovirus expressing DN-RSK (Ad.DN-RSK) and measured NHE1 activity. The baseline rate of pH recovery in acid-loaded cells was equal in cells expressing LacZ or DN-RSK. However, NHE1 activation by 100 micromol/L H2O2 was significantly inhibited in cells expressing DN-RSK (0.16+/-0.02 pH units/min) compared with Ad.LacZ (0.49+/ 0.13 pH units/min). Apoptosis induced by 12 hours of anoxia followed by 24 hours' reoxygenation was significantly reduced in cells expressing Ad.DN-RSK (18.6+/ 2.0%) compared with Ad.LacZ (29.3+/-5.4%). CONCLUSIONS: In summary, RSK is a novel regulator of cardiac NHE1 activity by phosphorylating NHE1 serine 703 and a new pathological mediator of I/R injury in the heart. PMID- 16717155 TI - Images in cardiovascular medicine. A swelling of the right neck and sudden death. PMID- 16717154 TI - Maturational and adaptive modulation of left ventricular torsional biomechanics: Doppler tissue imaging observation from infancy to adulthood. AB - BACKGROUND: Left ventricular (LV) torsional deformation, based in part on the helical myocardial fiber architecture, is an important component of LV systolic and diastolic performance. However, there is no comprehensive study describing its normal development during childhood and adult life. METHODS AND RESULTS: Forty-five normal subjects (25 children and 20 adults; aged 9 days to 49 years; divided into 5 groups: infants, children, adolescents, and young and middle-age adults) underwent assessment of LV torsion and untwisting rate by Doppler tissue imaging. LV torsion increased with age, primarily owing to augmentation in basal clockwise rotation during childhood and apical counterclockwise rotation during adulthood. Although LV torsion and untwisting overall showed age-related increases, when normalized by LV length, they showed higher values in infancy and middle age. The proportion of untwisting during isovolumic relaxation was lowest in infancy, increased during childhood, and leveled off thereafter, whereas peak untwisting performance (peak untwisting velocity normalized by peak LV torsion) showed a decrease during adulthood. CONCLUSIONS: We have shown the maturational process of LV torsion in normal subjects. Net LV torsion increases gradually from infancy to adulthood, but the determinants of this were different in the 2 age groups. The smaller LV isovolumic untwisting recoil during infancy and its decline in adulthood may suggest mechanisms for alterations in diastolic function. PMID- 16717156 TI - Images in cardiovascular medicine. Methemoglobinemia as a rare complication during a transesophageal echocardiogram. PMID- 16717158 TI - Letter regarding article by Khan et al, "prognostic value of troponin T and I among asymptomatic patients with end-stage renal disease: a meta-analysis". PMID- 16717160 TI - Cardiovascular disease management in Ireland. PMID- 16717161 TI - Rate control: is local better? PMID- 16717157 TI - Images in cardiovascular medicine. Multimodality imaging of cardiac sarcoidosis before and after steroid therapy. PMID- 16717162 TI - Reperfusion therapy starts in the ambulance. PMID- 16717163 TI - The dueling hazards of incomplete revascularization and incomplete data. PMID- 16717164 TI - Anemia in chronic heart failure: prevalence, etiology, clinical correlates, and treatment options. PMID- 16717165 TI - Ranolazine for the treatment of chronic angina and potential use in other cardiovascular conditions. PMID- 16717166 TI - The value of BNP testing. PMID- 16717167 TI - Atherosclerosis imaging of asymptomatic individuals: is the sales cart before the evidence horse? PMID- 16717168 TI - Noninvasive imaging of atherosclerosis among asymptomatic individuals. PMID- 16717169 TI - Accuracy of B-type natriuretic peptide tests to exclude congestive heart failure: systematic review of test accuracy studies. AB - BACKGROUND: Congestive heart failure (CHF) is a major public health problem. The use of B-type natriuretic peptide (BNP) tests shows promising diagnostic accuracy. Herein, we summarize the evidence on the accuracy of BNP tests in the diagnosis of CHF and compare the performance of rapid enzyme-linked immunosorbent assay (ELISA) and standard radioimmunosorbent assay (RIA) tests. METHODS: We searched electronic databases and the reference lists of included studies, and we contacted experts. Data were extracted on the study population, the type of test used, and methods. Receiver operating characteristic (ROC) plots and summary ROC curves were produced and negative likelihood ratios pooled. Random-effect meta analysis and metaregression were used to combine data and explore sources of between-study heterogeneity. RESULTS: Nineteen studies describing 22 patient populations (9 ELISA and 13 RIA) and 9093 patients were included. The diagnosis of CHF was verified by echocardiography, radionuclide scan, or echocardiography combined with clinical criteria. The pooled negative likelihood ratio overall from random-effect meta-analysis was 0.18 (95% confidence interval [CI], 0.13 0.23). It was lower for the ELISA test (0.12; 95% CI, 0.09-0.16) than for the RIA test (0.23; 95% CI, 0.16-0.32). For a pretest probability of 20%, which is typical for patients with suspected CHF in primary care, a negative result of the ELISA test would produce a posttest probability of 2.9%; a negative RIA test, a posttest probability of 5.4%. CONCLUSIONS: The use of BNP tests to rule out CHF in primary care settings could reduce demand for echocardiography. The advantages of rapid ELISA tests need to be balanced against their higher cost. PMID- 16717170 TI - Cost-effectiveness of B-type natriuretic peptide testing in patients with acute dyspnea. AB - BACKGROUND: B-type natriuretic peptide (BNP) is a quantitative marker of heart failure that seems to be helpful in its diagnosis. METHODS: We performed a prospective randomized study (B-Type Natriuretic Peptide for Acute Shortness of Breath Evaluation) including 452 patients who presented to the emergency department with acute dyspnea to estimate the long-term cost-effectiveness of BNP guidance. Participants were randomly assigned to a diagnostic strategy involving the measurement of BNP levels (n = 225) or assessment in a standard manner (n = 227). Nonparametric bootstrapping was used to estimate the distribution of incremental costs and effects on the cost-effectiveness plane during 180 days of follow-up. RESULTS: Testing of BNP induced several important changes in management of dyspnea, including a reduction in the initial hospital admission rate, the use of intensive care, and total days in the hospital at 180 days (median, 10 days [interquartile range, 2-24 days] in the BNP group vs 14 days [interquartile range, 6-27 days] in the control group; P = .005). At 180 days, all-cause mortality was 20% in the BNP group and 23% in the control group (P = .42). Total treatment cost was significantly reduced in the BNP group (7930 dollars vs 10,503 dollars in the control group; P = .004). Analysis of incremental 180-day cost-effectiveness showed that BNP guidance resulted in lower mortality and lower cost in 80.6%, in higher mortality and lower cost in 19.3%, and in higher or lower mortality and higher cost in less than 0.1% each. Results were robust to changes in most variables but sensitive to changes in rehospitalization with BNP guidance. CONCLUSION: Testing of BNP is cost-effective in patients with acute dyspnea. PMID- 16717171 TI - A brief measure for assessing generalized anxiety disorder: the GAD-7. AB - BACKGROUND: Generalized anxiety disorder (GAD) is one of the most common mental disorders; however, there is no brief clinical measure for assessing GAD. The objective of this study was to develop a brief self-report scale to identify probable cases of GAD and evaluate its reliability and validity. METHODS: A criterion-standard study was performed in 15 primary care clinics in the United States from November 2004 through June 2005. Of a total of 2740 adult patients completing a study questionnaire, 965 patients had a telephone interview with a mental health professional within 1 week. For criterion and construct validity, GAD self-report scale diagnoses were compared with independent diagnoses made by mental health professionals; functional status measures; disability days; and health care use. RESULTS: A 7-item anxiety scale (GAD-7) had good reliability, as well as criterion, construct, factorial, and procedural validity. A cut point was identified that optimized sensitivity (89%) and specificity (82%). Increasing scores on the scale were strongly associated with multiple domains of functional impairment (all 6 Medical Outcomes Study Short-Form General Health Survey scales and disability days). Although GAD and depression symptoms frequently co occurred, factor analysis confirmed them as distinct dimensions. Moreover, GAD and depression symptoms had differing but independent effects on functional impairment and disability. There was good agreement between self-report and interviewer-administered versions of the scale. CONCLUSION: The GAD-7 is a valid and efficient tool for screening for GAD and assessing its severity in clinical practice and research. PMID- 16717172 TI - The impact of prescribing safety alerts for elderly persons in an electronic medical record: an interrupted time series evaluation. AB - BACKGROUND: Considerable effort and attention have focused on medication safety in elderly persons; one approach that has been understudied in the outpatient environment is the use of computerized provider order entry with clinical decision support. The objective of this study was to examine the effects of computerized provider order entry with clinical decision support in reducing the use of potentially contraindicated agents in elderly persons. METHODS: With data from a 39-month period of a natural experiment, we evaluated changes in medication dispensing using interrupted time series analysis to estimate changes, controlling for prealert prescribing trends. The setting was a large health maintenance organization in the Pacific Northwest. All adult enrollees of the health plan participated. The intervention was computerized alerts cautioning against using certain medications in elderly persons. The main outcome measure was dispensing per 10,000 members per month. RESULTS: Following the implementation of the drug-specific alerts, a large and persistent reduction (5.1 prescriptions per 10,000, P=.004), a 22% relative decrease from the month before alert implementation, in the exposure of elderly patients to nonpreferred medications was observed. We found no evidence of a decrease in use of nonpreferred agents for nonelderly patients. The reduction seen in use of nonpreferred agents for elderly persons was driven primarily by decreases in dispensing for tertiary tricyclic agents. CONCLUSIONS: We found that alerts in an outpatient electronic medical record aimed at decreasing prescribing of medication use in elderly persons may be an effective method of reducing prescribing of contraindicated medications. The effect of the alerts on patient outcomes is less certain and deserves further investigation. PMID- 16717173 TI - Lowering the threshold for discussions of domestic violence: a randomized controlled trial of computer screening. AB - BACKGROUND: Women experiencing domestic violence (DV) frequent health care settings, but DV is rarely identified. METHODS: We conducted a randomized controlled trial to determine the effect of computer screening on health care provider-patient DV communication at 2 socioeconomically diverse emergency departments (EDs). Consenting nonemergent female patients, aged 18 to 65 years, were randomized to self-administered computer-based health risk assessment, with a prompt for the health care provider, or to "usual care"; all visits were audiotaped. Outcome measures were rates of DV discussion, disclosure, and services. RESULTS: Of 2169 eligible patients, 1281 (59%) consented; 871 (68%) were successfully audiotaped, and 903 (71%) completed an exit questionnaire. Rates of current DV risk on exit questionnaire were 26% in the urban ED and 21% in the suburban ED. In the urban ED, the computer prompt increased rates of DV discussion (147/262 [56%] vs 123/275 [45%]; P = .004), disclosure (37/262 [14%] vs 23/275 [8%]; P = .07), and services provided (21 [8%] vs 10 [4%]; P = .04). Women at the suburban site and those with private insurance or higher education were much less likely to be asked about experiences with abuse. Only 48% of encounters with a health care provider prompt regarding potential DV risk led to discussions. Both inquiries about and disclosures of abuse were associated with higher patient satisfaction with care. CONCLUSIONS: Computer screening for DV increased but did not guarantee that DV would be addressed during ED encounters. Nonetheless, it is likely that low-cost interventions that allow patients the opportunity to self-disclose can be used to improve detection of DV. PMID- 16717174 TI - Performance-based physical function and future dementia in older people. AB - BACKGROUND: The association of physical function with progression to dementia has not been well investigated. We aimed to determine whether physical function is associated with incident dementia and Alzheimer disease (AD). METHODS: We performed a prospective cohort study of 2288 persons 65 years and older without dementia. Patients were enrolled from 1994 to 1996 and followed up through October 2003. Main outcome measures included incident dementia and AD. RESULTS: During follow-up 319 participants developed dementia (221 had AD). The age specific incidence rate of dementia was 53.1 per 1000 person-years for participants who scored lower on a performance-based physical function test at baseline (< or = 10 points) compared with 17.4 per 1000 person-years for those who scored higher (> 10 points). A 1-point lower performance-based physical function score was associated with an increased risk of dementia (hazard ratio, 1.08; 95% confidence interval, 1.03-1.13; P < .001), an increased risk of AD (hazard ratio, 1.06; 95% confidence interval, 1.01-1.12; P = .01), and an increased rate of decline in the Cognitive Ability Screening Instrument scores (0.11 point per year; 95% confidence interval, 0.08-0.14; P < .001) after adjusting for age, sex, years of education, baseline cognitive function, APOE epsilon4 allele, family history of AD, depression, coronary heart disease, and cerebrovascular disease. CONCLUSIONS: Lower levels of physical performance were associated with an increased risk of dementia and AD. The study suggests that poor physical function may precede the onset of dementia and AD and higher levels of physical function may be associated with a delayed onset. PMID- 16717175 TI - Safety of high doses of influenza vaccine and effect on antibody responses in elderly persons. AB - BACKGROUND: Immune responses after influenza immunization are reduced in elderly individuals, the group at greatest risk for complications and death after influenza. Improved vaccines are needed to address this problem. METHODS: Ambulatory individuals 65 years and older (N = 202) were assigned randomly to receive a single intramuscular injection of the 2001-2002 formulation of trivalent inactivated influenza vaccine containing 15, 30, or 60 microg of hemagglutinin per strain (up to 180 microg total per dose) or placebo. Clinical and serologic responses were assessed during the month after immunization. RESULTS: Increasing dosages of vaccine elicited significantly higher serum antibody levels, frequencies of antibody responses, and putative protective titers after vaccination. Mean serum hemagglutination inhibition antibody titers 1 month after immunization in groups given 0-, 15-, 30-, and 60-microg dosages were 23, 37, 50, and 61 against influenza A/H1N1; 43, 86, 91, and 125 against influenza A/H3N2; and 10, 14, 18, and 24 against influenza B, respectively. Mean serum hemagglutination inhibition and neutralizing antibody levels against the 3 vaccine antigens in participants given the 60-microg dosage were 44% to 71% and 54% to 79%, respectively, higher than those in participants given the standard 15 microg dosage, and the 60-microg dosage level nearly doubled the frequency of antibody responses in those whose preimmunization antibody titers were in the lower half of the antibody range. Dose-related increases in the occurrence of injection site reactions were observed (P<.001), but all dosages were well tolerated. CONCLUSION: The improved immunogenicity of high-dose influenza vaccine among elderly persons should lead to enhanced protection against naturally occurring influenza. PMID- 16717176 TI - Management of urinary incontinence in Medicare managed care beneficiaries: results from the 2004 Medicare Health Outcomes Survey. AB - BACKGROUND: Despite the high prevalence of urinary incontinence (UI) among older persons and the existence of effective treatments, UI remains underreported by patients and underdiagnosed by clinicians. We measured the occurrence of UI problems in Medicare managed care beneficiaries, frequency of physician-patient communication regarding UI, and frequency of UI treatment. METHODS: We used cross sectional data from the 2004 Medicare Health Outcomes Survey, which measured self reported UI (accidental leakage of urine) and UI problems in the past 6 months, 36-Item Short-Form Health Survey health measures, discussions of UI with a health care provider, and receipt of UI treatment. RESULTS: The overall incidence of UI within the past 6 months was 37.3%, consistent with previous estimates. Problems with UI were strongly associated with poorer self-reported health. Mean 36-Item Short-Form Health Survey physical and mental health scores were lower by more than 5 points (on a 100-point scale, P<.001) for respondents with major UI problems when controlling for age, sex, race, Hispanic ethnicity, and major comorbidities. These differences were among the largest of any condition measured. Only 55.5% of those with self-reported UI problems reported discussing these problems during their recent visit to a physician or other health care provider. The rate of patient-reported UI treatment was 56.5% and was lower (P<.001) for older individuals (eg, 46.3% for those aged 90-94 years) or those with poor self-reported health status (50.5%). CONCLUSIONS: Among older persons, UI is common, underdiagnosed, and associated with substantial functional impairment. There appears to be considerable opportunity to mitigate the effects of UI on health and quality of life among community-dwelling older persons. PMID- 16717177 TI - Serum urea nitrogen, creatinine, and estimators of renal function: mortality in older patients with cardiovascular disease. AB - BACKGROUND: Renal dysfunction predicts increased mortality in cardiovascular patients, but the best renal estimator for quantifying risks is uncertain. We compared admission serum urea nitrogen (SUN) level, creatinine level, Modification of Diet in Renal Disease (MDRD) rate, and Mayo estimated glomerular filtration rate (eGFR) for predicting mortality. METHODS: In a retrospective cohort of Medicare patients (aged > or = 65 years) hospitalized for myocardial infarction (n = 44,437) and heart failure (n = 56,652), renal estimators were compared for linearity with 1-year mortality risk, magnitude of risk, and relative importance for predicting risk (percentage variance explained) in proportional hazards models. RESULTS: The SUN level, creatinine level, and Mayo eGFR had linear associations with mortality. These measures predicted steadily increased risk in patients who experienced a myocardial infarction with a SUN level greater than 17 mg/dL (> 6.1 mmol/L), a creatinine level greater than 1.0 mg/dL (> 88.4 micromol/L), and a Mayo eGFR of less than 100 mL/min per 1.73 m2; and in patients who experienced heart failure with a SUN level greater than 16 mg/dL (> 5.7 mmol/L), a creatinine level greater than 1.1 mg/dL (> 97.2 micromol/L), and a Mayo eGFR of 90 mL/min per 1.73 m2 or less. In contrast, the MDRD eGFR had a J-shaped association and failed to identify increased risks in 50.0% of patients who experienced a myocardial infarction (with an MDRD eGFR > 55 mL/min per 1.73 m2) and 60.0% of patients who experienced heart failure (with an MDRD eGFR > 44 mL/min per 1.73 m2). The SUN level and Mayo eGFR had the greatest magnitude of risks. In myocardial infarction and heart failure patients, adjusted mortality increased by 3% and 7%, respectively, per 5-U increase in SUN, and by 3% and 9%, respectively, per 10-U decrease in Mayo eGFR (P<.001), based on models including both renal measures. Of all the measures, SUN had the greatest magnitude of relative importance for predicting mortality. CONCLUSIONS: In older cardiovascular patients, SUN- and creatinine-based measures were powerful predictors of postdischarge mortality. Only MDRD eGFR was less adequate in quantifying risks for patients with mild impairment. Novel estimators, such as the Mayo eGFR, may play an important role in outcomes' prognostication for these patients. PMID- 16717178 TI - Breast cancer prevention: time for randomized controlled trials with statins. PMID- 16717179 TI - Clinical trials are required to prove the chemopreventive worth of statins. PMID- 16717180 TI - Prevention of hip fracture in elderly women with Alzheimer disease. PMID- 16717181 TI - Suppression of methamphetamine-seeking behavior by nicotinic agonists. AB - To understand the mechanism of methamphetamine (MAP) craving from the viewpoint of nicotinic acetylcholinergic transmission, we examined the responsible site of the brain for anticraving effects produced by nicotinic agonists by using a MAP self-administration paradigm in rats. Systemic nicotine and an acetylcholinesterase inhibitor, donepezil, attenuated the reinstatement of MAP seeking behavior by means of the activation of nicotinic acetylcholinergic receptors, but not muscarinic acetylcholinergic receptors, in the nucleus accumbens core, prelimbic cortex, amygdala, and hippocampus. Among these regions, with the exception of the hippocampus, we also found functional differences in this reinstatement. The nicotinic antagonist mecamylamine alone did not reinstate MAP-seeking behavior. These results suggest that the inactivation of nicotinic acetylcholinergic transmission may be an essential factor in the appearance of MAP-seeking behavior, and, thus, the normalization of the inactivated state may result in the suppression of the reinstatement. Our findings also indicate that there are functional differences in the responsible brain subregions. Extending this view to the treatment of MAP dependence, our results suggest that activators of nicotinic acetylcholinergic transmission are possible anticraving agents. PMID- 16717183 TI - Complex of linker histone H5 with the nucleosome and its implications for chromatin packing. AB - Linker histones are essential for chromatin filament formation, and they play key roles in the regulation of gene expression. Despite the determination of structures of the nucleosome and linker histones, the location of the linker histone on the nucleosome is still a matter of debate. Here we show by computational docking that the globular domain of linker histone variant H5 (GH5) has three distinct DNA-binding sites, through which GH5 contacts the DNA at the nucleosome dyad and the linker DNA strands entering and exiting the nucleosome. Our results explain the extensive mutagenesis and crosslinking data showing that side chains spread throughout the GH5 surface interact with nucleosomal DNA. The nucleosome DNA contacts positively charged side chains that are conserved within the linker histone family, indicating that our model extends to linker histone nucleosome interactions in general. Furthermore, our model provides a structural mechanism for formation of a dinucleosome complex specific to the linker histone H5, explaining its efficiency in chromatin compaction and transcription regulation. Thus, this work provides a basis for understanding how structural differences within the linker histone family result in functional differences, which in turn are important for gene regulation. PMID- 16717182 TI - Histamine signaling through the H(2) receptor in the Peyer's patch is important for controlling Yersinia enterocolitica infection. AB - Enteric pathogens such as Yersinia enterocolitica readily colonize and induce disease within the lymphatic tissues of the small intestine. To gain a comprehensive view of the host response to pathogens within these tissues, we determined the transcriptional profiles of intestinal lymphatic tissue infected with Y. enterocolitica. Expression analysis using Affymetrix GeneChips revealed a complex host response in the Peyer's patches and mesenteric lymph nodes after oral infection with Y. enterocolitica. Interestingly, histidine decarboxylase (Hdc) was significantly up-regulated in response to Y. enterocolitica infection. HDC is the enzyme solely responsible for the production of the biogenic amine histamine. Although histamine is well known for its role in allergy and for its effects on immunity and inflammation, little is known about its role or specific histamine receptors during the host response to bacterial infection. In this study, we provide evidence that histamine signaling through the histamine H(2) but not the H(1) receptor is important for controlling Y. enterocolitica infection within the Peyer's patches and mesenteric lymph nodes of mice. PMID- 16717185 TI - A missing model in reproductive skew theory: the bordered tug-of-war. AB - Models of reproductive skew can be classified into two groups: transactional models, in which group members yield shares of reproduction to each other in return for cooperation, and tug-of-war models, in which group members invest group resources in a tug-of-war over their respective reproductive shares. We synthesize these two models to yield a "bordered tug-of-war" model in which the internal tug-of-war is limited ("bordered") by the requirement that group members must achieve a certain amount of reproduction lest they pursue a noncooperative option leading to group breakup. Previous attempts to synthesize these two models did not allow for the fact that the tug-of-war will affect group output, which in turn feeds back on the reproductive payments required by group members to remain cooperative. The bordered tug-of-war model, which does not assume complete reproductive control by any individual and allows for conflict within groups, predicts that the degree of within-group selfishness will increase as the noncooperative options become less attractive, e.g., as ecological constraints on solitary breeding increase. When the noncooperative option involves fighting for the group resource (e.g., territory) and leaving if the fight is lost, the subordinate's overall share of reproduction is predicted to be independent of its relatedness to the dominant and to increase the greater its probability of winning the fight, the less the value of the territory, and the greater its personal payoff for leaving. The unique predictions of the bordered tug-of-war model may fit skew data from a number of species, including meerkats, lions, and wood mice. PMID- 16717184 TI - Liver-specific deletion of acetyl-CoA carboxylase 1 reduces hepatic triglyceride accumulation without affecting glucose homeostasis. AB - In animals, liver and white adipose are the main sites for the de novo fatty acid synthesis. Deletion of fatty acid synthase or acetyl-CoA carboxylase (ACC) 1 in mice resulted in embryonic lethality, indicating that the de novo fatty acid synthesis is essential for embryonic development. To understand the importance of de novo fatty acid synthesis and the role of ACC1-produced malonyl-CoA in adult mouse tissues, we generated liver-specific ACC1 knockout (LACC1KO) mice. LACC1KO mice have no obvious health problem under normal feeding conditions. Total ACC activity and malonyl-CoA levels were approximately 70-75% lower in liver of LACC1KO mice compared with that of the WT mice. In addition, the livers of LACC1KO mice accumulated 40-70% less triglycerides. Unexpectedly, when fed fat free diet for 10 days, there was significant up-regulation of PPARgamma and several enzymes in the lipogenic pathway in the liver of LACC1KO mice compared with the WT mice. Despite the significant up-regulation of the lipogenic enzymes, including a >2-fold increase in fatty acid synthase mRNA, protein, and activity, there was significant decrease in the de novo fatty acid synthesis and triglyceride accumulation in the liver. However, there were no significant changes in blood glucose and fasting ketone body levels. Hence, reducing cytosolic malonyl-CoA and, therefore, the de novo fatty acid synthesis in the liver, does not affect fatty acid oxidation and glucose homeostasis under lipogenic conditions. PMID- 16717186 TI - Developmental basis for hind-limb loss in dolphins and origin of the cetacean bodyplan. AB - Among mammals, modern cetaceans (whales, dolphins, and porpoises) are unusual in the absence of hind limbs. However, cetacean embryos do initiate hind-limb bud development. In dolphins, the bud arrests and degenerates around the fifth gestational week. Initial limb outgrowth in amniotes is maintained by two signaling centers, the apical ectodermal ridge (AER) and the zone of polarizing activity (ZPA). Our data indicate that the cetacean hind-limb bud forms an AER and that this structure expresses Fgf8 initially, but that neither the AER nor Fgf8 expression is maintained. Moreover, Sonic hedgehog (Shh), which mediates the signaling activity of the ZPA, is absent from the dolphin hind-limb bud. We find that failure to establish a ZPA is associated with the absence of Hand2, an upstream regulator of Shh. Interpreting our results in the context of both the cetacean fossil record and the known functions of Shh suggests that reduction of Shh expression may have occurred approximately 41 million years ago and led to the loss of distal limb elements. The total loss of Shh expression may account for the further loss of hind-limb elements that occurred near the origin of the modern suborders of cetaceans approximately 34 million years ago. Integration of paleontological and developmental data suggests that hind-limb size was reduced by gradually operating microevolutionary changes. Long after locomotor function was totally lost, modulation of developmental control genes eliminated most of the hind-limb skeleton. Hence, macroevolutionary changes in gene expression did not drive the initial reduction in hind-limb size. PMID- 16717187 TI - Alcohol-sensitive GABA receptors and alcohol antagonists. PMID- 16717188 TI - TL1A: a mediator of gut inflammation. PMID- 16717189 TI - Evolving stealth: genetic adaptation of Pseudomonas aeruginosa during cystic fibrosis infections. PMID- 16717190 TI - Thymic output in aged mice. AB - Using GFP to mark recent thymic emigrants (RTEs) in mice carrying a GFP transgene driven by the recombination-activating gene 2 promoter, we demonstrate that RTEs are readily detectable even in 2-year-old mice, despite the fact that the proportion of the peripheral T cell pool comprised of RTEs declines with age. Although the number of RTEs decreases after reaching a peak at 6 weeks of age, thymic output as a function of thymic size is surprisingly age-independent. The CD4:CD8 ratio of RTEs declines with age, partly because of a striking decrease in steady-state proliferation of CD4+ RTEs in older mice. RTEs in aged mice undergo phenotypic maturation in the lymphoid periphery with delayed kinetics compared with young mice. RTEs from aged mice secrete less IL-2, proliferate less well, and achieve only weak expression of early-activation markers compared with more mature naive peripheral T cells from the same mice. The proportion of GFP- cells in the CD4+ and CD8+ thymic compartments increases with age, partly as a result of leakiness in the aged thymus, allowing reentry of naive peripheral T cells. PMID- 16717191 TI - An S-nitrosothiol (SNO) synthase function of hemoglobin that utilizes nitrite as a substrate. AB - Red blood cells (RBCs) act as O(2)-responsive transducers of vasodilator and vasoconstrictor activity in lungs and tissues by regulating the availability of nitric oxide (NO). Vasodilation by RBCs is impaired in diseases characterized by hypoxemia. We have proposed that the extent to which RBCs constrict vs. dilate vessels is, at least partly, controlled by a partitioning between NO bound to heme iron and to Cysbeta93 thiol of hemoglobin (Hb). Hemes sequester NO, whereas thiols deploy NO bioactivity. In recent work, we have suggested that specific micropopulations of NO-liganded Hb could support the chemistry of S nitrosohemoglobin (SNO-Hb) formation. Here, by using nitrite as the source of NO, we demonstrate that a (T state) micropopulation of a heme-NO species, with spectral and chemical properties of Fe(III)NO, acts as a precursor to SNO-Hb formation, accompanying the allosteric transition of Hb to the R state. We also show that at physiological concentrations of nitrite and deoxyHb, a S nitrosothiol precursor is formed within seconds and produces SNO-Hb in high yield upon its prompt exposure to O(2) or CO. Deoxygenation/reoxygenation cycling of oxyHb in the presence of physiological amounts of nitrite also efficiently produces SNO-Hb. In contrast, high amounts of nitrite or delays in reoxygenation inhibit the production of SNO-Hb. Collectively, our data provide evidence for a physiological S-nitrosothiol synthase activity of tetrameric Hb that depends on NO-Hb micropopulations and suggest that dysfunction of this activity may contribute to the pathophysiology of cardiopulmonary and blood disorders. PMID- 16717192 TI - The RNA-binding protein Musashi is required intrinsically to maintain stem cell identity. AB - A key goal of regenerative medicine is an understanding of the genetic factors that define the properties of stem cells. However, stem cell research in mammalian tissue has been hampered by a paucity of stem cell-specific markers. Although increasing evidence suggests that members of the Musashi (Msi) family of RNA-binding proteins play important functions in progenitor cells, it remains unclear whether there is a stem cell-autonomous requirement for Msi because of an inability to distinguish stem cells from early-lineage cells in mammalian tissues. Here, using the Drosophila testis as a model system for the study of stem cell regulation, we show specific evidence for a cell-autonomous requirement for Msi family proteins in regulating stem cell differentiation, leading to the identification of an RNA-binding protein required for spermatogonial stem cell maintenance. We found that loss of Msi function disrupts the balance between germ line stem cell renewal and differentiation, resulting in the premature differentiation of germ-line stem cells. Moreover, we found that, although Msi is expressed in both somatic and germ cells, Msi function is required intrinsically in stem cells for maintenance of stem cell identity. We also discovered a requirement for Msi function in male meiosis, revealing that Msi has distinct roles at different stages of germ cell differentiation. We describe the complementary expression patterns of the murine Msi paralogues Msi1 and Msi2 during spermatogenesis, which support the idea of distinct, evolutionarily conserved roles of Msi. PMID- 16717193 TI - Cytosolic chaperonin protects folding intermediates of Gbeta from aggregation by recognizing hydrophobic beta-strands. AB - Cytosolic chaperonin containing t-complex polypeptide 1 (CCT)/TRiC is a group II chaperonin that assists in the folding of newly synthesized proteins. It is a eukaryotic homologue of the bacterial group I chaperonin GroEL. In contrast to the well studied functions of GroEL, the substrate recognition mechanism of CCT/TRiC is poorly understood. Here, we established a system for analyzing CCT/TRiC functions by using a reconstituted protein synthesis by using recombinant elements system and show that CCT/TRiC strongly recognizes WD40 proteins particularly at hydrophobic beta-strands. Using the G protein beta subunit (Gbeta), a WD40 protein that is very rich in beta-sheets, as a model substrate, we found that CCT/TRiC prevents aggregation and assists in folding of Gbeta, whereas GroEL does not. Gbeta has a seven-bladed beta-propeller structure; each blade is formed from a WD40 repeat sequence encoding four beta-strands. Detailed mutational analysis of Gbeta indicated that CCT/TRiC, but not GroEL, preferentially recognizes hydrophobic residues aligned on surfaces of beta strands in the second WD40 repeat of Gbeta. These findings indicate that one of the CCT/TRiC-specific targets is hydrophobic beta-strands, which are highly prone to aggregation. PMID- 16717194 TI - Microtubule affinity-regulating kinase 2 functions downstream of the PAR-3/PAR 6/atypical PKC complex in regulating hippocampal neuronal polarity. AB - The PAR-3/PAR-6/atypical PKC (aPKC) complex is required for axon-dendrite specification of hippocampal neurons. However, the downstream effectors of this complex are not well defined. In this article, we report a role for microtubule affinity-regulating kinase (MARK)/PAR-1 in axon-dendrite specification. Knocking down MARK2 expression with small interfering RNAs induced formation of multiple axon-like neurites and promoted axon outgrowth. Ectopic expression of MARK2 caused phosphorylation of tau (S262) and led to loss of axons, and this phenotype was rescued by expression of PAR-3, PAR-6, and aPKC. In contrast, the polarity defects caused by an MARK2 mutant (T595A), which is not responsive to aPKC, were not rescued by the PAR-3/PAR-6/aPKC complex. Moreover, polarity was abrogated in neurons overexpressing a mutant of MARK2 with a deleted kinase domain but an intact aPKC-binding domain. Finally, suppression of MARK2 rescued the polarity defects induced by a dominant-negative aPKC mutant. These results suggest that MARK2 is involved in neuronal polarization and functions downstream of the PAR 3/PAR-6/aPKC complex. We propose that aPKC in complex with PAR-3/PAR-6 negatively regulates MARK(s), which in turn causes dephosphorylation of microtubule associated proteins, such as tau, leading to the assembly of microtubules and elongation of axons. PMID- 16717195 TI - Alternative splicing in concert with protein intrinsic disorder enables increased functional diversity in multicellular organisms. AB - Alternative splicing of pre-mRNA generates two or more protein isoforms from a single gene, thereby contributing to protein diversity. Despite intensive efforts, an understanding of the protein structure-function implications of alternative splicing is still lacking. Intrinsic disorder, which is a lack of equilibrium 3D structure under physiological conditions, may provide this understanding. Intrinsic disorder is a common phenomenon, particularly in multicellular eukaryotes, and is responsible for important protein functions including regulation and signaling. We hypothesize that polypeptide segments affected by alternative splicing are most often intrinsically disordered such that alternative splicing enables functional and regulatory diversity while avoiding structural complications. We analyzed a set of 46 differentially spliced genes encoding experimentally characterized human proteins containing both structured and intrinsically disordered amino acid segments. We show that 81% of 75 alternatively spliced fragments in these proteins were associated with fully (57%) or partially (24%) disordered protein regions. Regions affected by alternative splicing were significantly biased toward encoding disordered residues, with a vanishingly small P value. A larger data set composed of 558 SwissProt proteins with known isoforms produced by 1,266 alternatively spliced fragments was characterized by applying the pondr vsl1 disorder predictor. Results from prediction data are consistent with those obtained from experimental data, further supporting the proposed hypothesis. Associating alternative splicing with protein disorder enables the time- and tissue-specific modulation of protein function needed for cell differentiation and the evolution of multicellular organisms. PMID- 16717196 TI - Engineered viral vaccine constructs with dual specificity: avian influenza and Newcastle disease. AB - Avian influenza viruses of the H5 and H7 hemagglutinin subtypes, and Newcastle disease virus (NDV), are important pathogens in poultry worldwide. Specifically, the highly pathogenic H5N1 avian influenza virus is a particular threat because it has now occurred in more than 40 countries on several continents. Inasmuch as most chickens worldwide are vaccinated with a live NDV vaccine, we embarked on the development of vaccine prototypes that would have dual specificity and would allow a single immunization against both avian influenza and Newcastle disease. Using reverse genetics, we constructed a chimeric avian influenza virus that expressed the ectodomain of the hemagglutinin-neuraminidase gene of NDV instead of the neuraminidase protein of the H5N1 avian influenza virus. Our second approach to creating a bivalent vaccine was based on expressing the ectodomain of an H7 avian influenza virus hemagglutinin in a fusogenic and attenuated NDV background. The insertion into the NDV genome of the foreign gene (containing only its ectodomain, with the transmembrane and cytoplasmic domains derived from the F protein of NDV) resulted in a chimeric virus with enhanced incorporation of the foreign protein into virus particles. A single immunization of chickens with this improved vaccine prototype virus induced not only a 90% protection against an H7N7 highly pathogenic avian influenza virus, but also complete immunity against a highly virulent NDV. We propose that chimeric constructs should be developed for convenient, affordable, and effective vaccination against avian influenza and Newcastle disease in chickens and other poultry. PMID- 16717198 TI - Patient page. More heads are better than one: stroke team offers best results. PMID- 16717197 TI - Newcastle disease virus expressing H5 hemagglutinin gene protects chickens against Newcastle disease and avian influenza. AB - Newcastle disease virus (NDV)-expressing avian influenza virus (AIV) hemagglutinin (HA) of subtype H5 was constructed by reverse genetics. A cloned full-length copy of the genome of the lentogenic NDV strain Clone 30 was used for insertion of the ORF encoding the HA of the highly pathogenic AIV isolate A/chicken/Italy/8/98 (H5N2) in the intergenic region between the NDV fusion and hemagglutinin-neuraminidase (HN) genes. Remarkably, two species of HA transcripts were detected in cells infected with the resultant NDVH5. In a second recombinant (NDVH5m), a NDV transcription termination signal-like sequence located within the HA ORF was eliminated by silent mutations. Consequently, NDVH5m produced 2.7-fold more full-length HA transcripts, expressed higher levels of HA, and also incorporated more HA protein into its envelope than NDVH5. NDVH5m stably expressed the modified HA gene for 10 egg passages and both recombinants were found innocuous after intracerebral inoculation of 1-day-old chickens. Immunization of chickens with NDVH5m induced NDV- and AIVH5-specific antibodies and protected chickens against clinical disease after challenge with a lethal dose of velogenic NDV or highly pathogenic AIV, respectively. Remarkably, shedding of influenza virus was not observed. Furthermore, immunization with NDVH5m permitted serological discrimination of vaccinated and AIV field virus infected animals based on antibodies against the nucleoprotein of AIV. Therefore, recombinant NDVH5m is suitable as a bivalent vaccine against NDV and AIV and may be used as marker vaccine for the control of avian influenza. PMID- 16717200 TI - MRI findings of rapidly progressive ophthalmoplegia and blindness in mucormycosis. PMID- 16717202 TI - The lumping and splitting of inflammatory CNS diseases. PMID- 16717201 TI - Lead toxicity, white matter lesions, and aging. PMID- 16717203 TI - If not pharmacology, maybe physics. PMID- 16717204 TI - A 3-year clinical follow-up of adult patients with 3243A>G in mitochondrial DNA. AB - OBJECTIVE: To follow the clinical course of patients with the mitochondrial DNA mutation 3243A>G for 3 years. METHODS: Thirty-three adult patients with the 3243A>G mutation entered a 3-year follow-up study. They were clinically evaluated annually, audiometry was performed, and samples were drawn for the analysis of blood chemistry and mutation heteroplasmy in leukocytes. Holter recording was performed three times during the follow-up and echocardiography, neuropsychological assessment, and quantitative EEG and brain imaging conducted at entry and after 3 years. RESULTS: The incidence of new neurologic events was low during the 3-year follow-up. Sensorineural hearing impairment (SNHI) progressed, left ventricular wall thickness increased, mean alpha frequency in the occipital and parietal regions decreased, and the severity of disease index (modified Rankin score) progressed significantly. The rate of SNHI progression correlated with mutation heteroplasmy in muscle. The increase in left ventricular wall thickness was seen almost exclusively in diabetic patients. Seven patients died during the follow-up, and they were generally more severely affected than those who survived. CONCLUSIONS: Significant changes in the severity of disease, sensorineural hearing impairment, left ventricular hypertrophy, and quantitative EEG were seen in adult patients with 3243A>G during the 3-year follow-up. PMID- 16717205 TI - Past adult lead exposure is linked to neurodegeneration measured by brain MRI. AB - OBJECTIVE: To determine whether cumulative lead dose in former organolead workers was associated with MRI measures of white matter lesions (WML) and global and structure-specific brain volumes. METHODS: MRIs, tibia lead, and other measures were obtained from 532 former organolead workers with a mean age of 56 years and a mean of 18 years since last occupational exposure to lead. Cumulative lead dose was measured by tibia lead, obtained by X-ray fluorescence, and expressed as microg lead per gram of bone mineral (microg Pb/g). WML were evaluated using the Cardiovascular Health Study grading scale. A total of 21 global and specific brain regions were evaluated. RESULTS: A total of 36% of individuals had WML grade of 1 to 7 (0 to 9 scale). Increasing peak tibia lead was associated with increasing WML grade (p = 0.004). The adjusted OR for a 1 microg Pb/g increase in tibia lead was 1.042 (95% CI = 1.021, 1.063) for a CHS grade of 5+ (> or = 5 vs < 5). In linear regression, the coefficient for tibia lead was negative for associations with all structures. Higher tibia lead was significantly related to smaller total brain volume, frontal and total gray matter volume, and parietal white matter volume. Of nine smaller specific regions of interest, higher tibia lead was associated with smaller volumes for the cingulate gyrus and insula. CONCLUSIONS: These data suggest that cumulative lead dose is associated with persistent brain lesions, and may explain previous findings of a progressive decline in cognitive function. PMID- 16717206 TI - Revised diagnostic criteria for neuromyelitis optica. AB - BACKGROUND: The authors previously proposed diagnostic criteria for neuromyelitis optica (NMO) that facilitate its distinction from prototypic multiple sclerosis (MS). However, some patients with otherwise typical NMO have additional symptoms not attributable to optic nerve or spinal cord inflammation or have MS-like brain MRI lesions. Furthermore, some patients are misclassified as NMO by the authors' earlier proposed criteria despite having a subsequent course indistinguishable from prototypic MS. A serum autoantibody marker, NMO-IgG, is highly specific for NMO. The authors propose revised NMO diagnostic criteria that incorporate NMO-IgG status. METHODS: Using final clinical diagnosis (NMO or MS) as the reference standard, the authors calculated sensitivity and specificity for each criterion and various combinations using a sample of 96 patients with NMO and 33 with MS. The authors used likelihood ratios and logistic regression analysis to develop the most practical and informative diagnostic model. RESULTS: Fourteen patients with NMO (14.6%) had extra-optic-spinal CNS symptoms. NMO-IgG seropositivity was 76% sensitive and 94% specific for NMO. The best diagnostic combination was 99% sensitive and 90% specific for NMO and consisted of at least two of three elements: longitudinally extensive cord lesion, onset brain MRI nondiagnostic for MS, or NMO-IgG seropositivity. CONCLUSIONS: The authors propose revised diagnostic criteria for definite neuromyelitis optica (NMO) that require optic neuritis, myelitis, and at least two of three supportive criteria: MRI evidence of a contiguous spinal cord lesion 3 or more segments in length, onset brain MRI nondiagnostic for multiple sclerosis, or NMO-IgG seropositivity. CNS involvement beyond the optic nerves and spinal cord is compatible with NMO. PMID- 16717207 TI - Effect of lamotrigine on cognition in children with epilepsy. AB - BACKGROUND: Lamotrigine does not affect cognition in healthy adult volunteers or adult patients with epilepsy, but its effect on cognition in children is uncertain. OBJECTIVE: To compare the effect of lamotrigine and placebo on cognition in children with well-controlled or mild epilepsy. METHOD: In a double blind, placebo-controlled, crossover study, 61 children with well-controlled or mild epilepsy were randomly assigned to add-on therapy with either lamotrigine followed by placebo or placebo followed by lamotrigine. Each treatment phase was 9 weeks, the crossover period 5 weeks. A neuropsychological test battery was performed during EEG monitoring at baseline and at the end of placebo and drug phases. The paired Student' t test was used for statistical analysis for neuropsychological data (two tailed) with a p value of 0.01 considered significant. Carryover and period effect were analyzed with generalized linear modeling (SPSS 10). RESULTS: Forty-eight children completed the study. Seizure frequency was similar during both treatment phases. No significant difference was found in continuous performance, binary choice reaction time, verbal and nonverbal recognition, computerized visual searching task, verbal and spatial delayed recognition, and verbal and nonverbal working memory between placebo and lamotrigine treatment phase. There was no significant carryover and period effect when corrected for randomization. CONCLUSION: Lamotrigine exhibits no clinically significant cognitive effects in adjunctive therapy for children with epilepsy. PMID- 16717208 TI - Elderly-onset essential tremor is associated with dementia. AB - OBJECTIVE: To determine whether essential tremor (ET) is associated with prevalent dementia. METHODS: The authors identified all persons with dementia and ET in a population-based study in central Spain (the Neurological Disorders in Central Spain [NEDICES] Study). Dementia was diagnosed using the Diagnostic and Statistical Manual of Mental Disorders (4th ed.) criteria. RESULTS: Thirty-one (11.4%) of 273 ET cases had dementia vs 204 (6.0%) of 3,382 non-ET subjects (controls) (odds ratio [OR] = 2.00, 95% CI = 1.34 to 2.98, p = 0.001). In a model that adjusted for age, stroke, and educational level, OR = 1.35, 95% CI = 0.87 to 2.18, and p = 0.17. In an adjusted model, ET cases with tremor onset after age 65 were 70% more likely to be demented than were controls (OR = 1.70, 95% CI = 1.04 to 2.76, p = 0.03), whereas ET cases with tremor onset at age < or = 65 and controls were equally likely to be demented (OR = 0.38, 95% CI = 0.09 to 1.73, p = 0.21). CONCLUSIONS: Older-onset essential tremor (ET) was associated with dementia. Prospective studies are required to elucidate the association between ET and dementia. PMID- 16717209 TI - Differential modulation of plasma beta-amyloid by insulin in patients with Alzheimer disease. AB - BACKGROUND: Hyperinsulinemia and insulin resistance are risk factors for memory impairment and Alzheimer disease (AD). Insulin regulates levels of the amyloid beta-peptide (Abeta) in vitro in neuronal cultures and in vivo in the CSF of normal older adults. OBJECTIVE: To determine whether insulin affected plasma Abeta levels and whether such effects differed for patients with AD compared with normal older adults. METHODS: Fifty-nine patients with AD and 50 healthy older adults each received infusions of saline and of insulin (1.0 mU.kg(-1).min(-1)) with accompanying dextrose to maintain euglycemia. A subset of participants (19 AD, 12 normal) received two additional conditions, in which insulin was infused at a lower (0.33 mU.kg(-1).min(-1)) and higher (1.67 mU.kg(-1).min(-1)) rate. Plasma insulin and Abeta were measured after 120 minutes of infusion. RESULTS: Adults with AD had higher plasma insulin vs normal adults at the two higher infusion rates, despite receiving comparable amounts of insulin. For normal adults, insulin reduced plasma Abeta levels at the middle (1.0 mU.kg(-1).min(-1)) dose, with attenuated effects at lower and higher doses. In contrast, for patients with AD, insulin raised plasma Abeta levels at the two higher doses (1.0 and 1.67 mU.kg(-1).min(-1)). CONCLUSIONS: These results suggest that patients with Alzheimer disease (AD) have reduced insulin clearance and insulin-provoked plasma amyloid beta-peptide (Abeta) elevation. Abnormal regulation of peripheral Abeta by insulin may contribute to AD risk. PMID- 16717210 TI - Characteristics of CADASIL in Korea: a novel cysteine-sparing Notch3 mutation. AB - OBJECTIVE: To elucidate the phenotype, genotype, and MRI findings of Korean patients with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) and mutation carriers. METHODS: The authors studied 40 members of nine unrelated Korean CADASIL families. After genetic analysis of Notch3, clinical and MRI findings were correlated in 27 mutation carriers. RESULT: Notch3 mutation sites were C174R (one family, n = 3), R133C (one family, n = 3), R587C (one family, n = 1), R544C (two families, n = 5), and R75P (four families, n = 15). The clinical features were typical of CADASIL, but the frequency of migraine in the Korean population appears low. MRI abnormalities were found in 54% of the mutant carriers, the most common being white matter hyperintensities. The prevalence of lacunes and microbleeds increased with patient age. Anterior temporal areas were less often involved in subjects with R75P mutations than in those where mutations occurred in other sites (p = 0.02). Gradient echo imaging identified microbleedings in 33% of mutation carriers (64% of those with abnormal MRI), whereas diffusion-weighted MRI showed abnormal findings in only one patient. Neurologic disability was related to the number of lacunar infarcts and the lesion volume of white matter hyperintensities (p < 0.001) whereas MMSE score was related to the number of lacunar infarcts (p < 0.005). CONCLUSIONS: Although Korean cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) mutation carriers show similar clinical and MRI findings, these abnormalities appear less frequently than in other populations. Relatively frequent microbleedings on gradient echo imaging suggest that treatment should be individualized according to MRI findings. The novel mutation of R75P, not involving a cysteine residue, is related to less frequent involvement of the anterior temporal area, thus broadening the spectrum of CADASIL. PMID- 16717211 TI - Brain volume changes in CADASIL: a serial MRI study in pure subcortical ischemic vascular disease. AB - BACKGROUND: Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an inherited small vessel disease causing stroke and subcortical vascular dementia. Recent studies in sporadic subcortical ischemic vascular disease have drawn attention to brain atrophy as a clinically important marker of disease progression. However, little is known about the role of brain atrophy and its clinical correlates in CADASIL. METHOD: The authors determined the normalized brain volume (NBV) and percent brain volume change (PBVC) over 2 years in 76 CADASIL subjects (45.1 +/- 9.7 years) using the SIENA (structural image evaluation using normalization of atrophy) software and its adaptation for cross-sectional measurements (SIENAX). Baseline assessments included systolic blood pressure (SBP), homocysteine levels, BMI, and APOE genotyping. T2-lesion volumes and clinical scales were assessed at both time points. RESULTS: The NBV significantly correlated with all clinical scores (Rankin, NIH Stroke Scale, Barthel, structured interview for the diagnosis of Alzheimer dementia and multi-infarct dementia, Mattis dementia rating scale) at both time points independently of age and sex. PBVC correlated with changes of all clinical scores (all p < 0.01) except for the Mattis dementia rating scale (p = 0.10). In a linear regression model, age (p < 0.001), male sex (p < 0.01), and SBP (p = 0.07) were the main risk factors for a lower NBV at baseline. Age (p < 0.001) and SBP (p = 0.01) were risk factors for brain volume loss during follow up. Sample size estimates showed that the number of individuals needed to demonstrate a treatment effect in a trial can be reduced when PBVC is used as an endpoint. CONCLUSIONS: This study identifies brain atrophy as an important aspect of the disease process in CADASIL and establishes significant correlations with multiple clinical aspects including cognition. Age and systolic blood pressure are risk factors for brain volume loss during follow-up. Percent brain volume change seems promising as an adjunct outcome measure in future interventional trials. PMID- 16717212 TI - The cognitive profiles of CADASIL and sporadic small vessel disease. AB - BACKGROUND: Interpretation of treatment trials in vascular dementia is confounded by the presence of coexistent Alzheimer disease (AD) pathology. The younger onset genetic disease cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) offers a model of pure vascular dementia, in which such confounding is unlikely. To validate CADASIL's use as a model it is important to show it results in a similar cognitive impairment. METHODS: The same neuropsychological assessment was administered to patients with CADASIL (n = 34, 14 of whom had had stroke), sporadic small vessel disease (SVD) presenting with lacunar stroke and having confluent leukoaraiosis (n = 54), and healthy controls (n = 25). RESULTS: A similar pattern of neuropsychological impairment was seen in the two diseases, with prominent early executive dysfunction. Patients with CADASIL and SVD performed worse than controls on Trails switching test (CADASIL p = 0.006; SVD p < 0.001), and on verbal fluency test (CADASIL p = 0.015; SVD p = 0.004). The SVD group also performed worse on immediate (p = 0.050) and delayed (p = 0.049) memory. When only patients with CADASIL with stroke were included in analysis with SVD subjects, all of whom had had stroke, a very similar cognitive profile was seen. The only difference was on verbal fluency, where CADASIL subjects performed worse (p = 0.044). CONCLUSION: Patients with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) and small vessel disease show a similar pattern of cognitive deficits. This suggests that CADASIL provides a model of pure vascular dementia relevant for sporadic small vessel disease vascular dementia. PMID- 16717213 TI - CT after subarachnoid hemorrhage: relation of cerebral perfusion to delayed cerebral ischemia. AB - BACKGROUND: Delayed cerebral ischemia (DCI) after subarachnoid hemorrhage (SAH) is difficult to predict. The authors studied the relation between several parameters of brain perfusion at admission and development of DCI. METHODS: The authors analyzed the admission CT perfusion (CTP) scans of 46 patients scanned within 72 hours after SAH. They assessed cerebral blood volume (CBV) and flow (CBF), mean transit time (MTT), and time to peak (TTP) for eight predefined regions of interest. For patients with and without DCI, the authors compared perfusion quantitatively and semiquantitatively. With receiver-operator characteristic (ROC) curves, the authors assessed the relationship between DCI and perfusion parameters. To assess the potential prognostic value, they calculated sensitivity and specificity of optimal threshold values for the semiquantitative data. RESULTS: DCI was not significantly related with quantitative perfusion values. For the semiquantitative data, patients with DCI had significantly more asymmetry in perfusion, and ROC curves indicated a good relation (0.75 to 0.81). Optimal threshold values distinguishing between patients with and without DCI were 0.77 for CBV and 0.72 for CBF ratios, and 0.87 seconds for MTT and 1.0 second for TTP differences. The corresponding sensitivity was 0.75 for all parameters; the specificity was 0.70 for CBV, 0.93 for CBF, 0.70 for MTT, and 0.90 for TTP. CONCLUSIONS: Delayed cerebral ischemia (DCI) is related to perfusion asymmetry on admission CT perfusion (CTP). The cerebral blood flow ratio (comparing contralateral regions of interest) seems the best prognosticator for development of DCI. Further studies are needed to investigate the additional value of CTP to other prognosticators for DCI and to validate the chosen threshold values. PMID- 16717215 TI - Intraplaque dissection of the basilar artery. PMID- 16717214 TI - Decreasing incidence of lacunar vs other types of cerebral infarction in a Japanese population. AB - BACKGROUND: There is scant information on secular trends in the incidence and survival of ischemic stroke subtypes. METHODS: The authors established three cohorts of Hisayama residents age > or =40 years in 1961 (1,618 subjects), 1974 (2,038 subjects), and 1988 (2,637 subjects). They followed up with each cohort for 12 years, comparing the incidence and survival rate of ischemic stroke subtypes. Morphologic examinations by autopsy or brain imaging was performed on most of the ischemic stroke cases in all cohorts. RESULTS: The age-standardized incidence of lacunar infarction significantly declined by 59% for men and by 28% for women from the first to the second cohort. It continued to decline by 41% for men, but the decline decelerated for women between the second and third cohort. The age-standardized incidence of atherothrombotic infarction tended to decline from the first to the second cohort, whereas it was sustained between the second and third cohort for both sexes. The age-standardized incidence of cardioembolic infarction was unchanged throughout the cohorts. In these cohorts, mean blood pressure levels among hypertensive subjects and the prevalence of current smoker decreased with time, though the prevalence of hypertension remained stable. The 5 year survival rate after lacunar infarction significantly improved among the cohorts, but those of atherothrombotic and cardioembolic infarction did not. CONCLUSIONS: These data suggest that, in the Japanese population, the incidence of lacunar infarction steadily declined for the last 40 years. The improvement of hypertension control and decreasing prevalence of smoking might be responsible for this trend. PMID- 16717216 TI - Aldose reductase inhibition alters nodal Na+ currents and nerve conduction in human diabetics. AB - BACKGROUND: In diabetic nerves, activation of the polyol pathway via an aldose reductase and the resulting impairment of the Na(+)-K(+) pump would lead to a decreased transaxonal Na+ gradient and thereby reduced nodal Na+ currents. OBJECTIVE: To investigate whether the aldose reductase inhibitor (ARI) epalrestat improves nodal Na+ currents and nerve conduction in human diabetic neuropathy. METHODS: The authors conducted a 6-month, open clinical trial with an ARI, epalrestat, in 30 patients with mild-to-moderate diabetic neuropathy. The latent addition technique and measurements of the strength-duration time constant were used to estimate nodal persistent Na+ currents in median motor axons. Excitability testing and extensive nerve conduction studies including F-wave analyses were performed before and 1 and 6 months after the initiation of treatment with oral epalrestat. RESULTS: Within a month of the start of treatment, there was a significant improvement in nerve conduction, particularly in conduction times across the carpal tunnel and F-wave latencies. The results of latent addition (p < 0.05) and strength-duration time constant (p = 0.06) suggested increased nodal persistent Na+ currents. At 6 months, nerve conduction continued to improve. CONCLUSIONS: Aldose reductase pathway inhibition could rapidly increase nodal Na+ currents and thereby improve the slowing of nerve conduction, presumably because of a restoration of the membranous Na+ gradient. PMID- 16717217 TI - Association between tPA therapy and raised early matrix metalloproteinase-9 in acute stroke. AB - BACKGROUND: Matrix metalloproteinase-9 (MMP9) is expressed in acute ischemic stroke and up-regulated by tissue plasminogen activator (tPA) in animal models. The authors investigated plasma MMP9 and its endogenous inhibitor, tissue inhibitor of metalloproteinase (TIMP1), in tPA-treated and -untreated stroke patients. METHODS: Nonstroke control subjects and consecutive ischemic stroke patients presenting within 8 hours of onset were enrolled. Blood was sampled within 8 hours and at 24 hours, 2 to 5 days and 4 to 6 weeks. MMP9 and TIMP1 were analyzed by ELISA and gel zymography. RESULTS: Fifty-two cases (26 tPA treated, 26 tPA untreated) and 27 nonstroke control subjects were enrolled. Hyperacute MMP9 was elevated in tPA-treated vs tPA-untreated patients (medians 43 vs 28 ng/mL; p = 0.01). tPA therapy independently predicted hyperacute MMP9 after adjustment for stroke severity, volume, and hemorrhagic transformation (p = 0.01). There was a trend toward lower hyperacute TIMP1 levels in tPA-treated vs tPA-untreated patients (p = 0.06). Hyperacute MMP9 was correlated to poor 3-month modified Rankin Scale outcome (r = 0.58, p = 0.0005). CONCLUSION: Tissue plasminogen activator independently predicted plasma matrix metalloproteinase-9 (MMP9) in the first 8 hours after human ischemic stroke. As MMP9 may be an important mediator of hemorrhagic transformation, alternative thrombolytic agents or therapeutic MMP9 inhibition may increase the safety profile of acute stroke thrombolysis. PMID- 16717218 TI - Preservation of cognitive function in dystonia treated by pallidal stimulation. AB - OBJECTIVE: To assess the effects of bilateral pallidal deep brain stimulation (DBS) on mood and cognitive performance in patients with dystonia before surgery (at baseline, while patients received their usual treatment) and 12 months postoperatively (while patients received neurostimulation and their medications) in a multicenter prospective study. METHODS: Twenty-two patients with primary generalized dystonia were evaluated with tests focused on executive functions. The authors considered the patients' severe disability and selected the following tests: Raven Progressive Matrices 38, Similarities and Arithmetic subtests of the Wechsler Adult Intelligence Scale-R, Grober and Buschke, Wisconsin Card Sorting Test (WCST), verbal fluency, Trail Making Test, and the Beck Depression Inventory. Median age at surgery was 30 years (range = 14 to 54 years), median duration of disease was 18.5 years (range = 4 to 37 years). RESULTS: Before surgery, no patients showed cognitive decline or depression. The surgical procedure appeared to be benign cognitively. One year after surgery, free recall improved. There was a significant reduction in the number of errors in the WCST. No behavioral or mood changes were found. CONCLUSIONS: Bilateral pallidal stimulation has a good benefit-to-risk ratio as it did not negatively affect cognitive performance and mood in primary dystonia, while a significant motor improvement was obtained. Moreover, a significant mild improvement in executive functions was observed, which may have been related either to the surgical treatment or to the marked decrease in anticholinergic drugs. PMID- 16717219 TI - Benjamin Franklin, electricity, and the palsies: on the 300th anniversary of his birth. AB - Benjamin Franklin was involved not only with the nature of electricity but with its possible medical utility. He conducted electrical experiments on people with palsies, notably those caused by stroke, to see if electricity from machines could restore movement. Franklin recognized that electricity was not the miraculous cure it was hoped to be, and he presented his findings in 1757 as communication to the Royal Society. Although he did not provide names or individual case studies in this communication, subsequently published in 1758, his personal letters reveal that he treated at least two important colonists: James Logan, William Penn's secretary and a prominent public official in Pennsylvania, and Jonathan Belcher, governor of several provinces. Franklin's private letters shed light on how he conducted his clinical "tryals" and why he drew the conclusions he did in his report to the Royal Society. PMID- 16717221 TI - Lumbar radiculopathy due to chondrocalcinosis. PMID- 16717220 TI - Expanding the clinical spectrum of POMT1 phenotype. AB - Mutations in POMT1 have been identified in Walker-Warburg syndrome and in patients with limb-girdle muscular dystrophy and mental retardation (LGMD2K). The authors report new POMT1 mutations in three unrelated children with severe motor impairment, leg hypertrophy, and mental retardation but without brain and ocular malformations. These patients are similar to LGMD2K but have earlier onset and more severe motor disability. The current findings expand the spectrum of POMT1 associated phenotypes. PMID- 16717222 TI - Neuromyelitis optica diagnosis in clinically isolated syndromes suggestive of multiple sclerosis. AB - The authors assessed the extent of overlap between current diagnostic criteria of neuromyelitis optica (NMO) and multiple sclerosis (MS) by applying NMO criteria to a large cohort of 320 patients with clinically isolated syndromes (CIS). Twenty-three (7.2%) patients fulfilled NMO absolute criteria at some time and 1 (0.3%) also fulfilled one major supportive criterion. Therefore, even by systematically applying NMO criteria in patients with CIS, NMO diagnosis is reached infrequently. PMID- 16717223 TI - Fixation-sensitive myoclonus in Lafora disease. AB - The authors report a patient with Lafora disease, whose myoclonus was suppressed by passive eye closure. Neurophysiologic studies disclosed that fixation was the most important enhancer of myoclonus. Magnetoencephalographic studies of visual evoked fields revealed abnormal activation of the visual corticocortical pathway via the insular cortex not seen in controls. The authors hypothesize that abnormal activation of the insular cortex may be involved in triggering the mechanism of fixation-sensitive myoclonus. PMID- 16717224 TI - Narcolepsy-cataplexy associated with precocious puberty. AB - In children, narcolepsy may be the symptom of a brain lesion or genetic disease. The authors report two cases with severe narcolepsy-cataplexy emerging in childhood in close temporal association with obesity and precocious puberty. PMID- 16717225 TI - Autosomal recessive ataxia with peripheral neuropathy and elevated AFP: novel mutations in SETX. AB - Mutations in the Senataxin gene (SETX) are associated with autosomal recessive ataxia-ocular apraxia 2 (AOA2) and autosomal dominant juvenile ALS (ALS4). Here, the authors describe novel homozygous missense mutations in SETX, M274I, and R1294C, found in two siblings with ataxia, peripheral neuropathy, and increased serum alpha-fetoprotein level and three other siblings with heterozygous missense mutations who were neurologically asymptomatic. The results demonstrate that the double missense mutations are responsible for AOA2 but not for ALS4. PMID- 16717226 TI - Phrenic neuropathy due to neuralgic amyotrophy. AB - The authors reviewed the medical records of 33 patients diagnosed with idiopathic phrenic neuropathy and found that 17 patients had clinical features of neuralgic amyotrophy. They concluded that a careful clinical and electrodiagnostic evaluation may implicate neuralgic amyotrophy as a causative disease in patients with apparently isolated phrenic neuropathy. PMID- 16717227 TI - Diagnostic evaluation of clinically normal subjects with chronic hyperCKemia. AB - The authors analyzed muscle biopsy specimens of 104 patients with creatine kinase activity greater than 500 UI/L (normal 10 to 170 UI/L) without signs of muscle weakness. They achieved a definite or probable diagnosis in 55% of cases. The most frequently identified diseases were glycogen storage diseases, muscular dystrophies, and inflammatory myopathies. The probability of making a diagnosis was higher in children and when creatine kinase level was greater than 2,000 UI/L. PMID- 16717228 TI - Genetic heterogeneity in paroxysmal nonkinesigenic dyskinesia. AB - Paroxysmal nonkinesigenic dyskinesia (PNKD) is characterized by attacks of dystonia or chorea lasting minutes to hours. Recently, mutations in the myofibrillogenesis regulator 1 gene (MR-1) have been identified in 10 unrelated PNKD kindreds. The authors describe a Canadian PNKD family who does not have mutations in the MR-1 gene and links to a separate locus at 2q31. This indicates that there are at least two different genes responsible for PNKD. PMID- 16717229 TI - Selective dendritic degeneration of medium spiny neurons in dementia with Lewy bodies. AB - Dementia with Lewy bodies (DLB) is the second most common form of dementia and shows more severely impaired performance on tests of executive functions compared to Alzheimer disease. Here the authors demonstrate selective spinodendritic degeneration of medium spiny neurons in regions of the caudate nucleus that subserve executive functions and propose that this may underlie, at least in part, the heightened executive dysfunction observed in patients with DLB. PMID- 16717230 TI - Late in-stent thrombosis following carotid angioplasty and stenting. AB - Acute in-stent thrombosis is a well-known complication of carotid angioplasty and stenting (CAS) and often due to resistance to or inadequate treatment with platelet-inhibiting agents. The authors report three cases of a delayed in-stent thrombosis after more than a week but less than 3 months after CAS. In all cases, the postprocedural antiplatelet regimen was discontinued to enable the treatment of a relevant comorbidity. PMID- 16717231 TI - Bilateral substantia nigra involvement in vaccine-associated poliomyelitis. PMID- 16717232 TI - Ocular neuromyotonia secondary to a cavernous sinus meningioma. PMID- 16717233 TI - Transient downbeat nystagmus from West Nile virus encephalomyelitis. PMID- 16717234 TI - Orthostatic tremor in monozygotic twins. PMID- 16717235 TI - Co-occurrence of a cavernous malformation and contralateral moyamoya. PMID- 16717236 TI - Reduced penetrance of intermediate size alleles in spinocerebellar ataxia type 10. PMID- 16717237 TI - The greatest generation. PMID- 16717238 TI - . . . soundtrack. . . PMID- 16717239 TI - Should the Babinski sign be part of the routine neurologic examination? PMID- 16717240 TI - Rate of progression differs in frontotemporal dementia and Alzheimer disease. PMID- 16717241 TI - CSF levels of angiogenesis-related proteins in patients with leptomeningeal metastases. PMID- 16717242 TI - Initial emergency department blood pressure as predictor of survival after acute ischemic stroke. PMID- 16717243 TI - Antiplatelet therapy contributes to acute deterioration of intracerebral hemorrhage. PMID- 16717244 TI - Long-term 24-hour duodenal infusion of levodopa: outcome and dose requirements. PMID- 16717245 TI - Serum autoantibodies do not differentiate PANDAS and Tourette syndrome from controls. PMID- 16717248 TI - Etiology of Parkinson's disease. AB - The etiology of Parkinson's disease (PD) has long been thought to involve both genetic and environmental factors, but until recently there has been no direct evidence to support either one as a causative factor. However, in the past 8 years six different genes have been identified as causing familial PD. Together, they support the notion that common pathogenetic mechanisms exist across the etiologic spectrum of PD. Specifically, mutations in alpha-synuclein, parkin, UCHL1, DJ1, PINK1, and LRRK2 cause PD, with a Mendelian pattern of inheritance. DJ1 and PINK1 are mitochondrial proteins and overexpression of alpha-synuclein and parkin induce mitochondrial defects. These same proteins are involved in the response to oxidative stress and affect proteasomal function. In contrast, few environmental factors have been characterized. Nevertheless, those toxins that have been demonstrated to have the ability to cause nigrostriatal cell death appear to interact by interfering with mitochondrial function, inducing oxidative stress, and modifying proteasomal function. Therefore, common themes are beginning to emerge in the etiopathogenesis of PD. This bodes well for research focused on the development of treatments that will modify the course of PD. PMID- 16717249 TI - The need for neuroprotective therapies in Parkinson's disease: a clinical perspective. AB - Slowing of disease progression remains a major unmet need in the treatment of Parkinson's disease (PD). Multiple factors are responsible for progression of disability in this disorder including worsening of cardinal motor features due to progressive nigral pathology, the evolution of poorly levodopa-responsive symptoms like freezing, postural instability and falls as well as motor complications of sustained treatment with levodopa. In addition, non-motor symptoms including cognitive decline, autonomic failure, sleep disorders and pain become increasingly prevalent with advancing disease and add to the overall burden of this disease. So far no treatment has been shown to significantly retard the progression of overall disability, and neuroprotective trials have been limited by design issues and a narrow focus on rates of decline of motor scores or imaging markers of nigrostriatal dysfunction only. Current evidence suggests that it may soon be possible to define populations at increased risk to develop PD and thus to target the "preclinical" phase of PD for neuroprotection. Such future trials will test intervention for their ability to prevent or retard the development of clinically overt PD in at-risk individuals. PMID- 16717250 TI - The pathogenesis of cell death in Parkinson's disease. AB - Concepts of pathogenesis in Parkinson's disease (PD) have been based on attempts to understand the mechanisms responsible for nigral dopaminergic cell death. Pathogenesis has been proposed to involve oxidative and nitrative stress, excitotoxicity, inflammation, mitochondrial dysfunction, and altered proteolysis. These processes are considered to form a complex cascade of interrelated events that lead to neuron death by way of apoptosis. However, current views on pathogenic mechanisms in PD may not be as exact as commonly proposed. Future concepts of pathogenesis in PD need to incorporate events leading to the destruction of non-dopaminergic nuclei and to distinguish between primary factors that are responsible for disease initiation and secondary factors that contribute to disease progression. Importantly, there is a need to determine whether PD is a single illness with a common pathogenesis or a group of related illnesses with different pathogenic mechanisms. This is an essential step to understanding pathogenesis and is critical to the development of comprehensive neuroprotective approaches to treatment. PMID- 16717251 TI - Proteasomal dysfunction in sporadic Parkinson's disease. AB - The cause and mechanism of neuronal death in sporadic Parkinson's disease (PD) continue to elude investigators. Recently, alterations in proteasomal function have been detected in the brain of patients with the illness. The biochemical basis of the defect and its relevance to the disease process are now being studied. The available results suggest that proteasomal dysfunction could underlie protein accumulation, Lewy body formation, and neuron death in PD. The cause of proteasomal dysfunction is unknown at present, but this could relate to gene mutations, oxidative damage, ATP depletion, or the actions of environmental toxins. It remains to be established if proteasomal dysfunction plays a primary or a secondary role in the initiation or progression of the neurodegenerative process in PD. PMID- 16717252 TI - Issues in neuroprotection clinical trials in Parkinson's disease. AB - Despite advances in understanding the pathogenesis of Parkinson's disease (PD), treatments that favorably influence the course of illness have remained elusive. In order to identify potentially neuroprotective interventions, improved clinical trial designs are needed. Researchers have to carefully consider what type of PD patients, how to measure the impact of putative neuroprotective agents, how to choose which interventions to study and what sequence of clinical trial designs is most appropriate. The possible use of futility studies to rapidly identify strongly ineffective agents and the use of delayed start designs to identify disease modifying interventions may both be important advances. Improved clinical trial design along with continued new insights into PD pathogenesis will likely lead to the identification of agents which can favorably influence the course of this disease. PMID- 16717253 TI - Clinical trials aimed at detecting neuroprotection in Parkinson's disease. AB - Potential neuroprotective therapies for Parkinson's disease (PD) are being identified in the laboratory and evaluated in the clinic in an effort to improve long-term outcomes for patients. Several clinical trial designs and methodologies have been used in an attempt to identify neuroprotective effects of medications. Such studies have evaluated (a) time to onset of a clinical milestone of disease progression, (b) progression of clinical symptoms from untreated baseline to an untreated endpoint obtained after wash-out of study intervention, (c) progression of clinical symptoms in early PD, (d) change in imaging markers over time, and (e) a combination of clinical (wash-out) and imaging markers. None of these approaches has yet provided a definitive means to evaluate neuroprotection. Clinical outcomes can be confounded by symptomatic effects of treatments, and imaging markers can be affected by pharmacologic or pharmodynamic changes resulting from treatment. Better methods of assessing putative neuroprotection in PD are needed. PMID- 16717254 TI - Rationale for considering that propargylamines might be neuroprotective in Parkinson's disease. AB - A neuroprotective therapy that slows or stops disease progression is the major unmet medical need in Parkinson's disease (PD). Current evidence indicates that cell death in PD occurs, at least in part, by way of a signal-mediated apoptotic process. This raises the possibility that anti-apoptotic agents might be neuroprotective in PD. Propargylamines have been demonstrated to be potent anti apoptotic agents in both in vitro and in vivo studies, presumably by maintaining glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a dimer and thereby preventing its nuclear translocation where it blocks upregulation of anti apoptotic proteins. Selegiline is a monamine oxidase type B (MAO-B) inhibitor that incorporates a propargyl ring within its molecular structure. It was shown to delay the need for symptomatic therapy in untreated PD patients in the DATATOP study, but interpretation is confounded by its symptomatic effects. Rasagiline is another MAO-B inhibitor that contains a propargyl ring and has protective effects in laboratory models. A clinical trial utilizing a delayed start design demonstrated that patients initiated on rasagiline at baseline are improved at one year in comparison to patients initiated on placebo and switched to rasagiline at 6 months even though both groups were on the same treatment for the last 6 months of the study. These results argue against the benefit being due to a symptomatic effect and are consistent with rasagiline having a protective effect. PMID- 16717255 TI - Clinical trials with rasagiline: evidence for short-term and long-term effects. AB - Rasagiline (N-propargyl-1 (R)-aminoindan) is a selective, potent irreversible inhibitor of MAO-B that possesses neuroprotective and anti-apoptotic properties in a variety of in vitro and in vivo animal models relevant to Parkinson's disease (PD). Several randomized controlled clinical trials have demonstrated the safety and efficacy of rasagiline as monotherapy in PD and as adjunctive therapy for patients receiving levodopa. In addition, the 1-year randomized, delayed start analysis of the TEMPO study suggests that rasagiline may slow the rate of progression of PD. The randomized delayed-start paradigm has potential to differentiate short-term symptomatic effects from long-term effects of anti parkinsonian agents. In the future, long-term trials to examine the potential disease-modifying effects of rasagiline, which incorporate biological markers as well as clinical endpoints, may further elucidate the role of rasagiline in the treatment of both early and advanced PD. PMID- 16717256 TI - Gene transfer of trophic factors and stem cell grafting as treatments for Parkinson's disease. AB - Current therapies for Parkinson's disease (PD) are limited in their ability to control PD symptomatology, are associated with motor and psychiatric side effects, and do not prevent disease progression. Considerable scientific and media interest has focused on the potential value of gene and stem cell therapies to overcome these problems and to enhance the quality of life for PD patients. Gene therapies utilize a viral vector to deliver a protein of interest to specific brain region. Clinical trials of gene therapy are currently underway using adeno-associated virus to deliver AADC to the striatum, the trophic factor nurturin to the striatum, and GAD to the STN. To date, no serious adverse effects have been noted, but only a small number of patients have been studied. Stem cells are pluripotential cells that offer the potential of generating unlimited numbers of optimized dopamine cells for transplantation. Stem cells can be grown and expanded in tissue culture and then induced to differentiate into dopamine neuronal phenotypes. Transplantation of these cells into the striatum is associated with behavioral improvement in 6-OHDA rodents and MPTP monkeys. Still, only small numbers of transplanted dopaminergic cells survive, and benefits are modest. Clinical trials in PD have not yet been performed. There is considerable enthusiasm for the potential of these procedures, but there remains much to learn in the laboratory and neither has been established to be effective as a treatment for PD. Long term safety and efficacy trials have not been performed in PD patients and the potential of unanticipated side effects must be addressed. Further, neither treatment is expected to improve the non-dopaminergic features of PD. PMID- 16717258 TI - Informed choice and deaf children: underpinning concepts and enduring challenges. AB - This article concerns the first stage of a research and development project that aimed to produce both parent and professional guidelines on the promotion and provision of informed choice for families with deaf children. It begins with a theoretical discussion of the problems associated with the concept of informed choice and deaf child services and then focuses specifically on why a metastudy approach was employed to address both the overcontextualized debate about informed choice when applied to deaf children and the problems associated with its investigation in practice with families and professionals. It presents a detailed analysis of the conceptual relevance of a range of identified studies "outside" the field of deafness. These are ordered according to 2 main conceptual categories and 7 subcategories-(a) the nature of information: "information that is evaluative, not just descriptive"; "the difficulties of information for a purpose"; "the origins and status of information"; and "informed choice and knowledge, not informed choice and information" and (b) parameters and definitions of choice: "informed choice as absolute and relative concept", "preferences and presumptions of rationality", and "informed choice for whom?" Relevant deaf child literature is integrated into the discussion of each conceptual debate in order both to expand and challenge current usage of informed choice as applied to deaf children and families and to delineate possible directions in the planning of the next stage of the main project aimed at producing parent/professional guidelines. PMID- 16717257 TI - Schizophrenia in translation: the presence of absence: habenular regulation of dopamine neurons and the encoding of negative outcomes. AB - Many patients with schizophrenia have pronounced deficits in the use of negative feedback to guide problem solving and learning, as seen on tasks like the Wisconsin Card Sorting Test. There is now a compelling body of evidence from nonhuman primates that suggests transient decreases in dopamine cell activity may reflect the occurrence of unexpected negative outcomes, such as the absence of an expected reward, and, generalizing to the human, the occurrence of negative feedback or the absence of expected reward. We present preliminary evidence that habenula projections to the midbrain are capable of producing a transient, but nearly complete, inhibition of dopamine neurons at a population level similar to that observed in behaving primates following an unexpected negative outcome. Human functional imaging studies offer further evidence that the habenula is activated following receipt of unexpected negative feedback or the absence of expected positive feedback. We present initial evidence that patients with schizophrenia lack appropriate modulation of habenula activity in response to feedback. Collectively, these data suggest that the habenula may play a critical role in mediating the feedback-processing deficits of schizophrenia. PMID- 16717259 TI - Eliminating barriers to DTC advertising for OTCs and natural health products. PMID- 16717260 TI - Monitoring the price of new drugs. PMID- 16717261 TI - Canada's future physicians: clinicians, researchers or teachers? PMID- 16717262 TI - Are there better ways to determine wait times? PMID- 16717263 TI - The lesser evils of battling round goby infiltration. PMID- 16717264 TI - An underrecognized cause of respiratory distress in a neonate. PMID- 16717265 TI - Getting physicians to accept new information technology: insights from case studies. AB - BACKGROUND: The success or failure of a computer information system (CIS) depends on whether physicians accept or resist its implementation. Using case studies, we analyzed the implementation of such systems in 3 hospitals to understand better the dynamics of physicians' resistance to CIS implementation. METHODS: We selected cases to maximize variation while allowing comparison of CIS implementations. Data were collected from observations, documentation and interviews, the last being the main source of data. Interviewees comprised 15 physicians, 14 nurses and 14 system implementers. Transcripts were produced; 45 segments of the transcripts were coded by several judges, with an appropriate level of intercoder reliability. We conducted within-case and cross-case analyses of the data. RESULTS: Initially, most staff were neutral or enthusiastic about the CIS implementations. During implementation, the level of resistance varied and in 2 instances became great enough to lead to major disruptions and system withdrawal. Implementers' responses to physicians' resistance behaviours played a critical role. In one case, the responses were supportive and addressed the issues related to the real object of resistance; the severity of resistance decreased, and the CIS implementation was ultimately successful. In the other 2 cases, the implementers' responses reinforced the resistance behaviours. Three types of responses had such an effect in these cases: implementers' lack of response to resistance behaviours, antagonistic responses, and supportive responses aimed at the wrong object of resistance. INTERPRETATION: The 3 cases we analyzed showed the importance of the roles played by implementers and users in determining the outcomes of a CIS implementation. PMID- 16717266 TI - Balancing the cyclooxygenase portfolio. PMID- 16717267 TI - Physicians will happily adopt information technology. PMID- 16717268 TI - Disabled medicine. PMID- 16717269 TI - Opioids for chronic noncancer pain: a meta-analysis of effectiveness and side effects. AB - BACKGROUND: Chronic noncancer pain (CNCP) is a major health problem, for which opioids provide one treatment option. However, evidence is needed about side effects, efficacy, and risk of misuse or addiction. METHODS: This meta-analysis was carried out with these objectives: to compare the efficacy of opioids for CNCP with other drugs and placebo; to identify types of CNCP that respond better to opioids; and to determine the most common side effects of opioids. We searched MEDLINE, EMBASE, CENTRAL (up to May 2005) and reference lists for randomized controlled trials of any opioid administered by oral or transdermal routes or rectal suppositories for CNCP (defined as pain for longer than 6 mo). Extracted outcomes included pain, function or side effects. Methodological quality was assessed with the Jadad instrument; analyses were conducted with Revman 4.2.7. RESULTS: Included were 41 randomized trials involving 6019 patients: 80% of the patients had nociceptive pain (osteoarthritis, rheumatoid arthritis or back pain); 12%, neuropathic pain (postherpetic neuralgia, diabetic neuropathy or phantom limb pain); 7%, fibromyalgia; and 1%, mixed pain. The methodological quality of 87% of the studies was high. The opioids studied were classified as weak (tramadol, propoxyphene, codeine) or strong (morphine, oxycodone). Average duration of treatment was 5 (range 1-16) weeks. Dropout rates averaged 33% in the opioid groups and 38% in the placebo groups. Opioids were more effective than placebo for both pain and functional outcomes in patients with nociceptive or neuropathic pain or fibromyalgia. Strong, but not weak, opioids were significantly superior to naproxen and nortriptyline, and only for pain relief. Among the side effects of opioids, only constipation and nausea were clinically and statistically significant. INTERPRETATION: Weak and strong opioids outperformed placebo for pain and function in all types of CNCP. Other drugs produced better functional outcomes than opioids, whereas for pain relief they were outperformed only by strong opioids. Despite the relative shortness of the trials, more than one-third of the participants abandoned treatment. PMID- 16717270 TI - Diabetes susceptibility. PMID- 16717271 TI - Inconsistent position on SSRI ads. PMID- 16717272 TI - Another proposal for primary care. PMID- 16717274 TI - Medical wait lists. PMID- 16717276 TI - Medical wait lists. PMID- 16717277 TI - Medical wait lists. PMID- 16717278 TI - Caenorhabditis elegans reporter fusion genes generated by seamless modification of large genomic DNA clones. AB - By determining spatial-temporal expression patterns, reporter constructs provide significant insights into gene function. Although additionally providing information on subcellular distribution, translational reporters, where the reporter is fused to the gene coding sequence, are used less frequently than simpler constructs containing only putative promoter sequences. Because these latter constructs may not contain all necessary regulatory elements, resulting expression patterns must be interpreted cautiously. To ensure inclusion of all such elements and provide details of subcellular localization, construction of translational reporters would, preferably, utilize genomic clones, containing the complete locus plus flanking regions and permit seamless insertion of the reporter anywhere within the gene. We have developed such a method based upon lambda Red-mediated recombineering coupled to a robust two-step counter-selection protocol. We have inserted either gfp or cfp precisely at the C-termini of three Caenorhabditis elegans target genes, each located within different fosmid clones, and examined previously with conventional reporter approaches. Resulting transgenic lines revealed reporter expression consistent with previously published data for the tagged genes and also provided additional information including subcellular distributions. This simple and straightforward method generates reporters highly likely to recapitulate endogenous gene expression and thus represents an important addition to the functional genomics toolbox. PMID- 16717279 TI - Genome prediction of PhoB regulated promoters in Sinorhizobium meliloti and twelve proteobacteria. AB - In proteobacteria, genes whose expression is modulated in response to the external concentration of inorganic phosphate are often regulated by the PhoB protein which binds to a conserved motif (Pho box) within their promoter regions. Using a position weight matrix algorithm derived from known Pho box sequences, we identified 96 putative Pho regulon members whose promoter regions contained one or more Pho boxs in the Sinorhizobium meliloti genome. Expression of these genes was examined through assays of reporter gene fusions and through comparison with published microarray data. Of 96 genes, 31 were induced and 3 were repressed by Pi starvation in a PhoB dependent manner. Novel Pho regulon members included several genes of unknown function. Comparative analysis across 12 proteobacterial genomes revealed highly conserved Pho regulon members including genes involved in Pi metabolism (pstS, phnC and ppdK). Genes with no obvious association with Pi metabolism were predicted to be Pho regulon members in S.meliloti and multiple organisms. These included smc01605 and smc04317 which are annotated as substrate binding proteins of iron transporters and katA encoding catalase. This data suggests that the Pho regulon overlaps and interacts with several other control circuits, such as the oxidative stress response and iron homeostasis. PMID- 16717280 TI - Downstream signaling mechanism of the C-terminal activation domain of transcriptional coactivator CoCoA. AB - The coiled-coil coactivator (CoCoA) is a transcriptional coactivator for nuclear receptors and enhances nuclear receptor function by the interaction with the bHLH PAS domain (AD3) of p160 coactivators. The C-terminal activation domain (AD) of CoCoA possesses strong transactivation activity and is required for the coactivator function of CoCoA with nuclear receptors. To understand how CoCoA AD transmits its activating signal to the transcription machinery, we defined specific subregions, amino acid motifs and protein binding partners involved in the function of CoCoA AD. The minimal transcriptional AD was mapped to approximately 91 C-terminal amino acids and consists of acidic, serine/proline rich and phenylalanine-rich subdomains. Transcriptional activation by the CoCoA AD was p300-dependent, and p300 interacted physically and functionally with CoCoA AD and was recruited to a promoter by the interaction with CoCoA AD. The FYDVASAF motif in the CoCoA AD was critical for the transcriptional activity of CoCoA AD, the interaction of CoCoA with p300, the coactivator function of CoCoA for estrogen receptor alpha and GRIP1 and the transcriptional synergy among coactivators GRIP1, CARM1, p300 and CoCoA. Taken together these data extend our understanding of the mechanism of downstream signaling by the essential C terminal AD of the nuclear receptor coactivator CoCoA; they indicate that p300 is a functionally important interaction partner of CoCoA AD and that their interaction potentiates transcriptional activation by the p160 coactivator complex. PMID- 16717281 TI - A modified version of a Fos-associated cluster in HBZ affects Jun transcriptional potency. AB - Like c-Fos, HBZ (HTLV-I bZIP factor) is able to interact with c-Jun but differs considerably from c-Fos in its ability to activate AP-1-responsive genes since HBZ rather inhibits transcriptional activity of c-Jun. To better understand the molecular mechanisms involved in this down-regulation of c-Jun activity, a large number of HBZ/c-Fos chimeras was constructed and analyzed for their ability to interact with c-Jun, to bind to the AP-1 motif and to stimulate expression of a reporter gene containing the collagenase promoter. By this approach, we demonstrate that the DNA-binding domain of HBZ is responsible for its inhibitory effect on the trans-activation potential of c-Jun. However, unexpectedly, we found that exchange of a cluster of six charged amino acids immediately adjacent to the DNA contact region altered significantly transcriptional activity of chimeras. This particular subdomain could be involved in efficient presentation of the AP-1 complex to the transcriptional machinery. To confirm this role, specific residues present in the cluster of HBZ were substituted for corresponding amino acids in c-Fos. Unlike the JunD-activating potential of wild type HBZ, this mutant was no longer able to stimulate JunD activity, confirming the key role of this particular cluster in regulation of Jun transcriptional potency. PMID- 16717282 TI - High potency silencing by single-stranded boranophosphate siRNA. AB - In RNA interference (RNAi), double-stranded short interfering RNA (ds-siRNA) inhibits expression from complementary mRNAs. Recently, it was demonstrated that short, single-stranded antisense RNA (ss-siRNA) can also induce RNAi. While ss siRNA may offer several advantages in both clinical and research applications, its overall poor activity compared with ds-siRNA has prevented its widespread use. In contrast to the poor gene silencing activity of native ss-siRNA, we found that the silencing activity of boranophosphate-modified ss-siRNA is comparable with that of unmodified ds-siRNA. Boranophosphate ss-siRNA has excellent maximum silencing activity and is highly effective at low concentrations. The silencing activity of boranophosphate ss-siRNA is also durable, with significant silencing up to 1 week after transfection. Thus, we have demonstrated that boranophosphate modified ss-siRNA can silence gene expression as well as native ds-siRNA, suggesting that boranophosphate-modified ss-siRNAs should be investigated as a potential new class of therapeutic agents. PMID- 16717283 TI - Multiple physical forms of excised group II intron RNAs in wheat mitochondria. AB - Plant mitochondrial group II introns do not all possess hallmark ribozymic features such as the bulged adenosine involved in lariat formation. To gain insight into their splicing pathways, we have examined the physical form of excised introns in germinating wheat embryos. Using RT-PCR and cRT-PCR, we observed conventional lariats consistent with a two-step transesterification pathway for introns such as nad2 intron 4, but this was not the case for the cox2 intron or nad1 intron 2. For cox2, we detected full-length linear introns, which possess non-encoded 3'terminaladenosines, as well as heterogeneous circular introns, which lack 3' nucleotide stretches. These observations are consistent with hydrolytic splicing followed by polyadenylation as well as an in vivo circularization pathway, respectively. The presence of both linear and circular species in vivo is supported by RNase H analysis. Furthermore, the nad1 intron 2, which lacks a bulged nucleotide at the branchpoint position, comprised a mixed population of precisely full-length molecules and circular ones which also include a short, discrete block of non-encoded nucleotides. The presence of these various linear and circular forms of excised intron molecules in plant mitochondria points to multiple novel group II splicing mechanisms in vivo. PMID- 16717284 TI - Target prediction for small, noncoding RNAs in bacteria. AB - Many small, noncoding RNAs in bacteria act as post-transcriptional regulators by basepairing with target mRNAs. While the number of characterized small RNAs (sRNAs) has steadily increased, only a limited number of the corresponding mRNA targets have been identified. Here we present a program, TargetRNA, that predicts the targets of these bacterial RNA regulators. The program was evaluated by assessing whether previously known targets could be identified. The program was then used to predict targets for the Escherichia coli RNAs RyhB, OmrA, OmrB and OxyS, and the predictions were compared with changes in whole genome expression patterns observed upon expression of the sRNAs. Our results show that TargetRNA is a useful tool for finding mRNA targets of sRNAs, although its rate of success varies between sRNAs. PMID- 16717285 TI - Structure-based prediction of insertion-site preferences of transposons into chromosomes. AB - Mobile genetic elements with the ability to integrate genetic information into chromosomes can cause disease over short periods of time and shape genomes over eons. These elements can be used for functional genomics, gene transfer and human gene therapy. However, their integration-site preferences, which are critically important for these uses, are poorly understood. We analyzed the insertion sites of several transposons and retroviruses to detect patterns of integration that might be useful for prediction of preferred integration sites. Initially we found that a mathematical description of DNA-deformability, called V(step), could be used to distinguish preferential integration sites for Sleeping Beauty (SB) transposons into a particular 100 bp region of a plasmid [G. Liu, A. M. Geurts, K. Yae, A. R. Srinivassan, S. C. Fahrenkrug, D. A. Largaespada,J. Takeda, K. Horie, W. K. Olson and P. B. Hackett (2005) J. Mol. Biol., 346, 161-173 ]. Based on these findings, we extended our examination of integration of SB transposons into whole plasmids and chromosomal DNA. To accommodate sequences up to 3 Mb for these analyses, we developed an automated method, ProTIS, that can generate profiles of predicted integration events. However, a similar approach did not reveal any structural pattern of DNA that could be used to predict favored integration sites for other transposons as well as retroviruses and lentiviruses due to a limitation of available data sets. Nonetheless, ProTIS has the utility for predicting likely SB transposon integration sites in investigator-selected regions of genomes and our general strategy may be useful for other mobile elements once a sufficiently high density of sites in a single region are obtained. ProTIS analysis can be useful for functional genomic, gene transfer and human gene therapy applications using the SB system. PMID- 16717286 TI - Yeast Protein Interactome topology provides framework for coordinated functionality. AB - The architecture of the network of protein-protein physical interactions in Saccharomyces cerevisiae is exposed through the combination of two complementary theoretical network measures, betweenness centrality and 'Q-modularity'. The yeast interactome is characterized by well-defined topological modules connected via a small number of inter-module protein interactions. Should such topological inter-module connections turn out to constitute a form of functional coordination between the modules, we speculate that this coordination is occurring typically in a pairwise fashion, rather than by way of high-degree hub proteins responsible for coordinating multiple modules. The unique non-hub-centric hierarchical organization of the interactome is not reproduced by gene duplication-and divergence stochastic growth models that disregard global selective pressures. PMID- 16717287 TI - Defining characteristics of Tn5 Transposase non-specific DNA binding. AB - While non-specific DNA plays a role in target localization for many recombinases, transcription factors and restriction enzymes, the importance of non-specific DNA interactions for transposases has not been investigated. Here, we discuss non specific DNA-Tn5 Transposase (Tnp) interactions and suggest how they stabilize the Tnp and modulate Tnp localization of the 19 bp Tnp recognition end sequences (ESes). DNA protection assays indicate that full-length Tnp interacts efficiently with supercoiled DNA that does not contain ESes. These interactions significantly prolong the lifetime of Tnp, in vitro. The balance between non-specific DNA bound and free Tnp is affected by DNA topology, yet, intermolecular transfer of active Tnp occurs with both supercoiled and linear non-specific DNA. Experiments with substrates of varying lengths show that Tn5 Tnp can utilize non-specific DNA to facilitate localization of an intramolecular ES over distances less than 464 bp. Finally, synaptic complex formation is inhibited in the presence of increasing concentrations of supercoiled and linear pUC19. These experiments strongly suggest that Tn5 Tnp has a robust non-specific DNA binding activity, that non specific DNA modulates ES sequence localization within the global DNA, most likely through a direct transfer mechanism, and that non-specific DNA binding may play a role in the cis bias manifested by Tn5 transposition. PMID- 16717290 TI - Reimbursement for cancer treatment: coverage of off-label drug indications. AB - Approximately half of the uses of anticancer chemotherapy drugs are for indications other than those referenced in the United States Food and Drug Administration approved label. Some managed care organizations and private health insurance plans have declined to reimburse the cost of drugs used off-label to treat cancer on the ground that these uses are "experimental" or "investigational." Cancer patients and their providers have experienced similar problems in the Medicare and Medicaid program. To a large extent, these issues have been addressed through legislation enacted in 1993 that requires coverage of medically appropriate cancer therapies including off-label uses recognized by established drug compendia and peer-reviewed literature. Congress has fashioned a system that has worked well, as reflected in improvements in cancer morbidity and mortality. Now, however, after more than a decade of success, the system requires attention. This statement of policy from the American Society of Clinical Oncology encourages the Secretary of the United States Department of Health and Human Services to address these unmet needs in order to ensure that patients with cancer have access to clinically appropriate treatment, as reflected in timely compendia listings and reports of studies in the medical literature. PMID- 16717289 TI - American Society of Clinical Oncology guideline for antiemetics in oncology: update 2006. AB - PURPOSE: To update the 1999 American Society of Clinical Oncology guideline for antiemetics in oncology. UPDATE METHODOLOGY: The Update Committee completed a review and analysis of data published from 1998 thru February 2006. The literature review focused on published randomized controlled trials, and systematic reviews and meta-analyses of published phase II and phase III randomized controlled trials. RECOMMENDATIONS: The three-drug combination of a 5 hydroxytryptamine-3 (5-HT(3)) serotonin receptor antagonist, dexamethasone, and aprepitant is recommended before chemotherapy of high emetic risk. For persons receiving chemotherapy of high emetic risk, there is no group of patients for whom agents of lower therapeutic index are appropriate first-choice antiemetics. These agents should be reserved for patients intolerant of or refractory to 5-HT3 serotonin receptor antagonists, neurokinin-1 receptor antagonists, and dexamethasone. The three-drug combination of a 5-HT3 receptor serotonin antagonist, dexamethasone, and aprepitant is recommended for patients receiving an anthracycline and cyclophosphamide. For patients receiving other chemotherapy of moderate emetic risk, the Update Committee continues to recommend the two-drug combination of a 5-HT3 receptor serotonin antagonist and dexamethasone. In all patients receiving cisplatin and all other agents of high emetic risk, the two drug combination of dexamethasone and aprepitant is recommended for the prevention of delayed emesis. The Update Committee no longer recommends the combination of a 5-HT3 serotonin receptor antagonist and dexamethasone for the prevention of delayed emesis after chemotherapeutic agents of high emetic risk. CONCLUSION The Update Committee recommends that clinicians administer antiemetics while considering patients' emetic risk categories and other characteristics. PMID- 16717288 TI - Disparate requirements for the Walker A and B ATPase motifs of human RAD51D in homologous recombination. AB - In vertebrates, homologous recombinational repair (HRR) requires RAD51 and five RAD51 paralogs (XRCC2, XRCC3, RAD51B, RAD51C and RAD51D) that all contain conserved Walker A and B ATPase motifs. In human RAD51D we examined the requirement for these motifs in interactions with XRCC2 and RAD51C, and for survival of cells in response to DNA interstrand crosslinks (ICLs). Ectopic expression of wild-type human RAD51D or mutants having a non-functional A or B motif was used to test for complementation of a rad51d knockout hamster CHO cell line. Although A-motif mutants complement very efficiently, B-motif mutants do not. Consistent with these results, experiments using the yeast two- and three hybrid systems show that the interactions between RAD51D and its XRCC2 and RAD51C partners also require a functional RAD51D B motif, but not motif A. Similarly, hamster Xrcc2 is unable to bind to the non-complementing human RAD51D B-motif mutants in co-immunoprecipitation assays. We conclude that a functional Walker B motif, but not A motif, is necessary for RAD51D's interactions with other paralogs and for efficient HRR. We present a model in which ATPase sites are formed in a bipartite manner between RAD51D and other RAD51 paralogs. PMID- 16717291 TI - Powerful prognostic stratification by [18F]fluorodeoxyglucose positron emission tomography in patients with metastatic breast cancer treated with high-dose chemotherapy. AB - PURPOSE: This study examines the use of [(18)F]fluorodeoxyglucose positron emission tomography (FDG-PET) for the evaluation of the therapeutic response for patients treated with high-dose chemotherapy (HDC) with autologous stem cell transplantation for metastatic breast cancer (MBC) focusing on prognostic stratification. PATIENTS AND METHODS: Forty-seven patients with MBC were treated with a maximum of three cycles of HDC. Therapeutic response was assessed with conventional imaging (CImg; including a computed tomography in all cases and ultrasound, mammography, and bone scanning as clinically indicated) and by FDG PET study performed after the last cycle of HDC. Parameters analyzed for predicting survival were FDG-PET and CImg results, pattern of disease, prior treatment, and HDC regimen. RESULTS: Complete responses were observed in 16 patients (37%) with CImg and 34 patients (72%) with FDG-PET. The FDG-PET result was the most powerful and independent predictor of survival; patients with a negative post-treatment FDG-PET had a longer median survival than patients with a positive FDG-PET (24 months v 10 months; P < .001). By multivariate analysis the relative risk (RR) of death was higher in patients with FDG-PET-positive disease (RR, 5.3), prior anthracycline treatment (RR, 3.3), or with visceral metastasis (RR, 2.4). CONCLUSION: A single FDG-PET study performed after completion of HDC for MBC can powerfully stratify for survival. This may have implications for how we should assess outcome after conventional-dose therapy for MBC and warrants additional study. PMID- 16717292 TI - Bone marrow transplantation versus prolonged intensive chemotherapy for children with acute lymphoblastic leukemia and an initial bone marrow relapse within 12 months of the completion of primary therapy: Children's Oncology Group study CCG 1941. AB - PURPOSE: To compare conventional sibling bone marrow transplantation (CBMT), BMT with alternative donor (ABMT), and chemotherapy (CT) for children with acute lymphoblastic leukemia (ALL) and an early first marrow relapse. PATIENTS AND METHODS: After informed consent, 214 patients with ALL and early marrow relapse began multiagent induction therapy. One hundred sixty-three patients with fewer than 25% marrow blasts and count recovery at the end of induction (second remission [CR2]) were allocated by donor availability. Fifty patients with sibling donors were allocated to CBMT. Seventy-two patients were randomly allocated between ABMT and CT while 41 patients refused allocation. RESULTS: Overall, 3-year event free survival from entry is 19% +/- 3%. Thirty-two of 50 CBMT patients (64%) and 19 of 37 ABMT patients (51%) underwent transplantation in CR2 with 3-year disease-free survival of 42% +/- 7% and 29% +/- 7%. The 3-year DFS is 29% +/- 7%, 21% +/- 7%, and 27% +/- 8% for patients allocated to CBMT, ABMT, and CT, respectively. Contrary to protocol, 12 of 35 patients allocated to CT underwent BMT in CR2. Of these, five patients died after BMT and 5 patients relapsed. CONCLUSION: More than one half of patients died, failed reinduction, or relapsed again before 3 months after CR2 (median time to BMT). Intent-to-treat pair-wise comparison of ABMT with CT, CT with CBMT, and CBMT with ABMT yields hazards of 1.2, 1.1, 0.8 with P values of .56, .80, and .36, respectively. Outcomes remain similar and poor for children with ALL and early marrow relapse. BMT is not a complete answer to the challenge of ALL and early marrow relapse. PMID- 16717293 TI - Phase I/II study of cetuximab in combination with cisplatin or carboplatin and fluorouracil in patients with recurrent or metastatic squamous cell carcinoma of the head and neck. AB - PURPOSE: This was an open, randomized, multicenter, phase I/II study to investigate the safety and tolerability of cetuximab in the first-line treatment of recurrent/metastatic squamous cell carcinoma of the head and neck (SCCHN). PATIENTS AND METHODS: Treatment comprised cetuximab (initial dose 400 mg/m2 with subsequent weekly doses of 250 mg/m2) in combination with 3-week cycles of either cisplatin (100 mg/m2) or carboplatin (area under the curve, 5), each in combination with a 5-day infusion of fluorouracil (FU) at escalating doses of 600, 800, and 1,000 mg/m2/d. The study was divided into two phases: A, the first two cycles (6 weeks) focusing on the safety and tolerability of combination therapy; and B, the remaining time for those benefiting from therapy until disease progression or intolerable toxicity. RESULTS: Fifty-three patients were enrolled onto the study. The incidence of dose-limiting toxicities in phase A was acceptable. The most common grade 3/4 adverse events in both groups were leucopenia (38%), asthenia (25%), vomiting (14%), and thrombocytopenia (15%), which are consistent with the known safety profiles of cetuximab, cisplatin/carboplatin, and FU. The overall response rate among patients was 36%, with no clear trend toward an increased efficacy at the highest dose of FU, and no impact of the concomitant chemotherapy regimens on cetuximab pharmacokinetics. CONCLUSION: The combination of cetuximab, cisplatin/carboplatin, and FU was reasonably well tolerated and active in recurrent/metastatic SCCHN, and merits additional investigation. An FU dose of 1,000 mg/m2/d in combination with cisplatin or carboplatin can be recommended for additional studies. PMID- 16717295 TI - Preexisting cognitive impairment in women before cardiac surgery and its relationship with C-reactive protein concentrations. AB - Preoperative cognitive state is seldom considered when investigating the effects of cardiac surgery on cognition. In this study we sought to determine the prevalence of cognitive impairment in women scheduled for cardiac surgery using nonhospitalized volunteers as a reference group and to examine the relationship between C-reactive protein levels and cognitive impairment. Psychometric testing was performed in 108 postmenopausal women scheduled for cardiac surgery and in 58 nonhospitalized control women. High sensitivity C-reactive protein levels were measured in the surgical patients. Preoperative cognitive impairment was defined as >2 sd lower scores on > or =2 tests compared with the controls. Cognitive impairment was present in 49 of 108 (45%) patients. C-reactive protein levels were higher for patients with compared with those without cognitive impairment (median, 8.1 mg/L versus 4.7 mg/L; P = 0.04). Based on multivariate logistic regression analysis, patient age, lower attained level of education, type 2 diabetes mellitus, and prior myocardial infarction identified risk for cognitive impairment (P < 0.05) but C-reactive protein levels did not (P = 0.09). In conclusion, cognitive impairment is prevalent in women before cardiac surgery. C reactive protein levels are increased in women with this condition but the relationship between this inflammatory marker and preexisting cognitive impairment is likely secondary to the acute phase reactant serving as a marker for other predisposing conditions. PMID- 16717296 TI - The marked reduction in mixed venous oxygen saturation during early mobilization after cardiac surgery: the effect of posture or exercise? AB - Early mobilization after cardiac surgery induces a marked reduction in mixed venous oxygen saturation (Svo2). Using pulmonary artery catheters and indirect calorimetry, we investigated the effects of exercise and postural change on cardiac index (CI) and Svo2 before and on the first morning after coronary artery bypass surgery. Sixteen patients with an ejection fraction >0.50 were studied at rest, during supine bicycle exercise, and during passive standing. Supine cycling at 30 W increased CI by 1.5 +/- 0.8 L x min(-1) x m(-2) before and 0.9 +/- 0.7 L x min(-1) x m(-2) after surgery (P < 0.05), whereas Svo2 was reduced from 80% +/- 4% at rest to 63 +/- 6% preoperatively (P < 0.05) and from 71% +/- 5% to 46% +/- 11% postoperatively (P < 0.05). Passive standing reduced CI by 0.8 +/- 0.5 L x min(-1) x m(-2) before and 0.3 +/- 0.4 L x min(-1) x m(-2) after surgery (P < 0.05). Svo2 was reduced from 79% +/- 5% to 64% +/- 7% preoperatively (P < 0.05) and from 72% +/- 6% to 60% +/- 6% postoperatively (P < 0.05). The exercise challenge revealed an altered cardiovascular response after surgery, causing a larger reduction in Svo2 for the same workload. Passive standing significantly reduced Svo2 both days, but this effect was less pronounced after surgery. The response to postural change and exercise was altered after surgery and may both contribute to the reduction in Svo2 during postoperative mobilization. PMID- 16717298 TI - Increased interleukin-6 after cardiac surgery predicts infection. AB - Early diagnosis and treatment of infection after cardiac surgery with cardiopulmonary bypass (CPB) improves outcome. Conventional laboratory tests, such as C-reactive protein and white blood cell count can not distinguish patients with early infection from those with systemic inflammatory response syndrome but without infection. After CPB, there is a systemic release of proinflammatory and antiinflammatory cytokines, including tumor necrosis factor alpha, interleukin (IL)-6, and IL-10. We investigated the predictive ability of these variables for infection after cardiac surgery. Forty-six patients with impaired left ventricular ejection fraction (<60%), scheduled for cardiac surgery, were included. Plasma samples were drawn 1 day before and immediately before surgery, on admission to the intensive care unit, and on days 1, 3, and 7 after surgery. Infection was identified according to the criteria of the Centers for Disease Control and Prevention. After surgery 13 patients developed an infection. In patients with infection, confirmed a median of 4 days after surgery, all measurements of IL-6, and IL-10 on postoperative day 3 were significantly increased. Tumor necrosis factor-alpha, leukocytes, and C-reactive protein were not increased in these patients. Immediately after surgery blood glucose was significantly increased in patients with infection. Increased IL-6 after CPB is predictive of infection after cardiac surgery in patients with impaired left ventricular function. PMID- 16717297 TI - Autoregulation of human jejunal mucosal perfusion during cardiopulmonary bypass. AB - Animal studies have suggested that autoregulation of intestinal blood flow is severely impaired during cardiopulmonary bypass (CPB). We investigated the jejunal mucosal capacity to autoregulate perfusion during nonpulsatile CPB (34 degrees C) in 10 patients undergoing elective cardiac surgery. Changes in mean arterial blood pressure (MAP) were induced by altering the CPB flow rate randomly for periods of 3 min from 2.4 L/min/m2 to either 1.8 or 3.0 L/min/m2. Jejunal mucosal perfusion (JMP) was continuously recorded by laser Doppler flowmetry. A typical pattern of flow motion (vasomotion) was recorded in all patients during CPB. Variations in CPB flow rates caused no significant changes in mean JMP, jejunal mucosal hematocrit, or red blood cell velocity within a range of MAP from 50 +/- 15 to 74 +/- 16 mm Hg. The vasomotion frequency and amplitude was positively correlated with CPB flow rate. IV injections of prostacyclin (10 microg, Flolan) blunted vasomotion and increased JMP from 192 +/- 53 to 277 +/- 70 (P < 0.05) perfusion units despite a reduction in MAP from 59 +/- 12 to 45 +/- 10 mm Hg (P < 0.05). Prostacyclin-induced vasodilation resulted in loss of mucosal autoregulation (pressure-dependent perfusion). We conclude that autoregulation of intestinal mucosal perfusion is maintained during CPB in humans. PMID- 16717299 TI - Endothelial Ca2+ signal transduction is altered by postoperative serum from patients undergoing coronary surgery with cardiopulmonary bypass. AB - Endothelial dysfunction after surgery may be caused by alterations in the intracellular signaling properties of endothelial cells. Functional alterations are believed to be systemic and dependent on the amount of invasiveness. This led us to assume that there would be a mediator in the blood. Therefore, we investigated the influence of perioperative serum obtained from patients undergoing highly invasive surgical interventions (cardiac surgery [CS] with cardiopulmonary bypass) and less invasiveness (total joint arthroplasty [TJA]) on endothelial single cell Ca2+ responses. Aortic endothelial cells were incubated with preoperative and postoperative serum samples from 26 patients undergoing CS and from 15 patients undergoing TJA. Adenosine triphosphate (100 microM)-induced alterations in FURA-2 fluorescence was used to measure intracellular Ca2+ in single cells. In CS samples the induced [Ca2+]i signals were enhanced by postoperative serum (peak levels: 96 +/- 41 FU versus 116 +/- 45 FU; P < 0.05). These postoperative enhancements were absent in TJA patients serum. Preincubation of CS samples with nifedipine to block voltage gated Ca2+ channels did not alter this effect, but the absence of extracellular Ca2+ abolished the increased response from postoperative CS serum exposure. Ca2+ entry probed with Mn2+ quenching was increased in endothelial cells exposed with postoperative CS serum and Ca2+ entry correlated with postoperative circulating interleukin-6 levels (P < 0.007). Endothelial functional alterations after CS with cardiopulmonary bypass are attributable, in part, to systemic factors present in serum that lead to specific endothelial enhanced Ca2+-signaling. This enhancement can be separated in vitro as an increased Ca2+ entry not present in serum from patients recovering from TJA. PMID- 16717300 TI - N-acetylcysteine for the prevention of kidney injury in abdominal aortic surgery: a randomized, double-blind, placebo-controlled trial. AB - In this prospective, randomized, placebo-controlled, double-blind trial we studied the effects of IV N-acetylcysteine for prevention of renal injury in patients undergoing abdominal aortic surgery. Seventy patients without previously documented renal dysfunction were randomly allocated to receive either N acetylcysteine (150 mg/kg mixed in 250 mL of 5% dextrose infused in 20 min, followed by an infusion of 150 mg/kg in 250 mL of 5% dextrose over 24 h) or placebo. The infusion was started after the induction of anesthesia. The primary outcome measure was renal injury as measured by the increases in urinary N-acetyl beta-d-glucosaminidase (NAG)/creatinine ratio (indicator of renal tubular injury) and urinary albumin/creatinine ratio (indicator of glomerular injury). Renal function was assessed by measuring plasma creatinine and serum cystatin C concentrations. The urinary NAG/creatinine ratio increased significantly from baseline to before crossclamp and remained increased on day 5 in both groups. The urinary albumin/creatinine ratio increased significantly from baseline to 6 h after declamping in the N-acetylcysteine group. However, the changes in the NAG/creatinine ratio and the albumin/creatinine ratio were not significantly different between the two groups. Plasma creatinine and serum cystatin C values remained unchanged during the study period in both groups. In conclusion, N acetylcysteine did not offer any significant protection from renal injury during elective aortic operation in patients with normal preoperative renal function, and some degree of tubular injury seems to occur before aortic crossclamp. PMID- 16717301 TI - A comparison of three phosphodiesterase type III inhibitors on mechanical and metabolic function in guinea pig isolated hearts. AB - Little is known about of the comparative cardiac lusitropic and coronary vasoactive effects of type III phosphodiesterase inhibitors independent of their systemic circulatory effects. We hypothesized that phosphodiesterase inhibitors have dissimilar concentration-dependent effects on cardiac function and metabolism and that their coronary vasodilatory effects are solely dependent on flow autoregulation secondary to positive inotropic effects. Our aim was to compare the dose-response electrophysiologic, mechanical, vasodilatory, and metabolic properties of three clinically available phosphodiesterase inhibitors in isolated Langendorff perfused guinea pig hearts. We found that, over a range from 10(-7) to 10(-4) M, amrinone, enoximone, and milrinone each produced maximal concentration-dependent positive chronotropic (12%, 18%, 26%), inotropic (16%, 26%, 26%), and lusitropic (14%, 21%, 19%) effects. At clinical concentrations, all phosphodiesterase inhibitors increased heart rate, but only milrinone significantly enhanced contractility and relaxation (11%). Each phosphodiesterase inhibitor similarly increased contractility at its highest concentration; this was accompanied by an increase in oxygen consumption, which was matched by comparable increases in coronary flow and oxygen delivery. Coronary flow reserve was preserved at the highest concentration of each drug, indicating that an increased metabolic rate was responsible for the increase in coronary flow by each drug at each concentration. Over the concentrations examined, we conclude that each of the phosphodiesterase inhibitors does not directly promote coronary vasodilation and that milrinone has the most prominent effects on contractility and relaxation at clinically relevant concentrations. PMID- 16717302 TI - The usefulness of transesophageal echocardiography during intraoperative cardiac arrest in noncardiac surgery. AB - According to guidelines established by the American Society of Anesthesiologists and the Society of Cardiovascular Anesthesiologists, life-threatening hemodynamic disturbances are classified as a category I indication for the intraoperative use of transesophageal echocardiography (TEE). However, the usefulness of TEE during intraoperative cardiac arrest and its impact on patient management have not been rigorously investigated. Using our departmental TEE database, we identified a population of 22 patients who underwent noncardiac surgical procedures and experienced unexpected intraoperative hemodynamic collapse requiring the initiation of Advanced Cardiac Life Support procedures between the time of induction of general anesthesia and the termination of the surgical procedure. Results of TEE examinations, patient records, detailed operative records, and outcome of patients were reviewed for the utility of TEE to diagnose the etiology of the hemodynamic collapse. Furthermore, the impact on subsequent patient management was evaluated. A primary suspected diagnosis of the underlying pathological process was established in 19 of 22 patients with TEE, including 9 with thromboembolic events, 6 with acute myocardial ischemia, 2 with hypovolemia, and 2 patients with pericardial tamponade. A definitive diagnosis could not be made in 3 patients with TEE. In 18 patients, TEE guided specific management beyond implementation of Advanced Cardiac Life Support protocols, including the addition of surgical procedures in 12 patients. Fourteen patients survived to leave the operating room, and 7 of these patients were eventually discharged from the hospital. Thus, TEE may provide additional diagnostic information in patients with intraoperative cardiac arrest and may directly guide specific, potentially life-saving therapy. PMID- 16717303 TI - Acute intracardiac thrombus formation during thoracoabdominal aortic surgery. PMID- 16717304 TI - Sinus of valsalva aneurysm obstructing the right ventricular outflow tract. PMID- 16717305 TI - Development of acute opioid tolerance during infusion of remifentanil for pediatric scoliosis surgery. AB - We tested the hypothesis that continuous intraoperative infusion of remifentanil is associated with the development of clinically relevant acute opioid tolerance in adolescents undergoing scoliosis surgery. Thirty adolescents were randomly assigned to receive an intraoperative analgesic regimen consisting of continuous remifentanil infusion or intermittent morphine alone. Postoperative analgesic consumption was assessed with a patient-controlled analgesia device that was used to self-administer morphine. Cumulative postoperative morphine consumption, pain scores, and sedation scores were recorded by a blinded investigator every hour for the first 4 h postoperatively and then every 4 h for a total of 24 h. Cumulative morphine consumption in the remifentanil group was significantly more than that in the morphine group at each time point in the initial 24 h after surgery (P < 0.0001). At 24 h after surgery, cumulative morphine consumption was 30% greater in the remifentanil group (1.65 +/- 0.41 mg/kg) than in the morphine group (1.27 +/- 0.32 mg/kg) (95% confidence interval for the difference, 0.11 to 0.65 mg/kg). Differences in pain and sedation scores were not statistically significant. These data suggest that intraoperative infusion of remifentanil is associated with the development of clinically relevant acute opioid tolerance in adolescents undergoing scoliosis surgery. PMID- 16717306 TI - Intraoperative administration of tramadol for postoperative nurse-controlled analgesia resulted in earlier awakening and less sedation than morphine in children after cardiac surgery. AB - In adults, intraoperative administration of tramadol could result in earlier recovery and less sedation than morphine. In this controlled, randomized, double blind study, we investigated whether an intraoperative initial dose of tramadol could cause more rapid awakening from general anesthesia, less sedation, and earlier tracheal extubation than morphine in children during the immediate postoperative period. Forty children aged 1-6 yr, scheduled for atrial or ventricular septal defect repair and tracheal extubation in the pediatric intensive care unit, were randomly allocated to receive morphine, initial dose 0.2 mg/kg, or tramadol 2 mg/kg given at the end of sternal closure, followed by nurse-controlled analgesia (bolus 0.02 mg/kg of morphine and 0.2 mg/kg of tramadol) with background infusions (0.015 mg x kg(-1) x h(-1) for morphine and 0.15 mg x kg(-1) x h(-1) for tramadol). Postoperatively, children receiving tramadol had earlier awakening from general anesthesia (P = 0.02) and were less sedated at 1 and 2 h postoperatively (P = 0.03 and P = 0.01, respectively). Tracheal extubation was earlier in the tramadol group (P = 0.01). Lengths of pediatric intensive care unit stay did not differ between groups. Times to first trigger of nurse-controlled analgesia bolus and objective pain scores during the 48 h observation period were comparable between groups. The incidence of desaturation and emesis were similar between groups. The patients ate well and did not differ on Day 1 or Day 2. PMID- 16717307 TI - Brain magnetic resonance imaging increases core body temperature in sedated children. AB - An increasing number of children now undergo magnetic resonance imaging (MRI) under sedation. MRI requires a cool environment. Because children have a larger surface area to body weight ratio than adults and because active warming devices are not MRI compatible, hypothermia as a result of passive heat loss is a risk. Absorption of radiofrequency radiation generated by the scanning process, however, may partially offset this heat loss. To determine the effect of absorbed radiofrequency radiation on body temperature during MRI, we measured pre-MRI and post-MRI tympanic temperatures in 30 children who underwent brain MRI while sedated with chloral hydrate and covered with a hospital gown and blanket. The mean (+/- sd) age was 14.9 +/- 8.6 mo, and weight was 9.8 +/- 2.8 kg. During an average scan duration of 42 +/- 13 min, mean tympanic temperatures increased 0.5 degrees C from 36.9 degrees C +/- 0.4 degrees C to 37.4 degrees C +/- 0.3 degrees C; (95% CI difference, 0.3 degrees C to 0.7 degrees C; P < 0.001). Our findings suggest that children sedated with chloral hydrate for brain MRI did not become hypothermic but rather had increased body temperature despite minimal barriers to heat loss and no active warming. These results imply that aggressive measures to prevent passive heat loss during MRI studies may not be needed in all patients. PMID- 16717308 TI - Ultrasonographic-guided ilioinguinal/iliohypogastric nerve block in pediatric anesthesia: what is the optimal volume? AB - Recently, our study group demonstrated the usefulness of ultrasonographic guidance in ilioinguinal/iliohypogastric nerve blocks in children. As a consequence, we designed a follow-up study to evaluate the optimal volume of local anesthetic for this regional anesthetic technique. Using a modified step-up step-down approach, with 10 children in each study group, a starting dose of 0.2 mL/kg of 0.25% levobupivacaine was administered to perform an ilioinguinal/iliohypogastric nerve block under ultrasonographic guidance. After each group of 10 patients, the results were analyzed, and if all blocks were successful, the volume of local anesthetic was decreased by 50%, and a further 10 patients were enrolled into the study. Failure to achieve a 100% success rate within a group subjected patients to an automatic increase of half the previous volume reduction to be used in the subsequent group. Using 0.2 and 0.1 mL/kg of 0.25% levobupivacaine, the success rate was 100%. With a volume of 0.05 mL/kg of 0.25% levobupivacaine, 4 of 10 children received additional analgesia because of an inadequate block. Therefore, according to the protocol, the amount was increased to 0.075 mL/kg of 0.25% levobupivacaine, where the success rate was again 100%. We conclude that ultrasonographic guidance for ilioinguinal/iliohypogastric nerve blocks in children allowed a reduction of the volume of local anesthetic to 0.075 mL/kg. PMID- 16717309 TI - Video assessment of supraglottic airway orientation through the perilaryngeal airway in pediatric patients. AB - The COBRA perilaryngeal airway (PLA) is a new supraglottic airway device available for use in children. We studied the orientation of the larynx as viewed through the PLA using video obtained with a fiberoptic bronchoscope in 45 infants and children. Laryngeal view was scored, and position of the grill bars of the PLA and the epiglottis was assessed. An acceptable airway was obtained in all subjects, but the laryngeal view was nearly or completely obstructed in 76.9% of the patients 10 kg and less, with the epiglottis folded over the glottic opening. This was uncommon in larger children. The grill bars of the PLA were closely opposed to the epiglottis and supraglottic structures in nearly all subjects. We conclude that 1) the PLA provides an acceptable airway in infants and children, but infolding of the epiglottis with obstruction of the view of the glottic opening is common in infants, 2) extra vigilance for airway obstruction in this age group is necessary, and 3) the PLA's close opposition to the supraglottic structures suggests that removal in a deeper plane of anesthesia might minimize laryngeal stimulation. PMID- 16717310 TI - Central nervous system electrical synapses as likely targets for intravenous general anesthetics. PMID- 16717311 TI - General anesthetics inhibit gap junction communication in cultured organotypic hippocampal slices. AB - Gap junctions are protein channels that directly connect the cytosol of neighboring cells, thus forming electrical synapses and promoting synchronous neuronal activities. Such activities lead to the initiation and propagation of electroencephalogram oscillations implicated in cognition and consciousness. In this study, we investigated the effects of propofol, thiopental, and halothane on gap junction communication in cultured organotypic hippocampal slices by recovery of fluorescence after photo bleaching (FRAP) technique and electrophysiological recordings. Propofol 15 microM and thiopental 10 microM attenuated gap junction communication in slice cultures by 46.7% +/- 4.5% and 48.8% +/- 5.5%, respectively, as measured by FRAP. Smaller concentrations of propofol 5 microM and thiopental 2 microM did not change gap junction coupling. Accompanying the decreased gap junction communication, hippocampus slice cultures exposed to propofol 15 microM and thiopental 10 microM were found to have reduced electrophysiologic spontaneous discharges and primary after discharges evoked by a tetanic train of 50 Hz for 2 s. On the other hand, halothane 0.64 mM, a concentration slightly larger than twice its minimum alveolar concentration had no effect on gap junction coupling while halothane 2.8 mM blocked FRAP by 70%. The current study illustrates that anesthetic concentrations of propofol and thiopental, but not halothane, attenuate gap junction communication in cultured hippocampal slices. Suppression of gap junction function could compound the mechanisms of anesthetic actions. PMID- 16717312 TI - Onset time for sevoflurane/nitrous oxide induction in adults is prolonged with increasing age. AB - Inhaled induction of anesthesia is occasionally used in adults. Using a modified vital capacity sevoflurane/nitrous oxide (N2O) inhaled induction, we evaluated the effect of increasing age on the onset time of anesthesia. Twenty patients, aged 26-65 yr, performed a vital capacity breath followed by regular tidal breathing from an anesthesia circuit primed with sevoflurane 8%/N2O/O2. The following values were recorded: time to loss of eyelash reflex (LOER); time to bispectral index < or =60 (BIS < or =60); expired fraction of sevoflurane at the time of induction, LOER and BIS < or =60. The mean times and 95% confidence intervals to LOER and BIS < or =60 were 54 s (37-70 s) and 175 s (143-207 s), respectively, and were significantly prolonged by aging (r = 0.65; P = 0.002). Times to LOER and BIS < or =60, predicted from the regression line, were 3.9 and 2 times longer in a 60-yr-old than in a 30-yr-old patient. The expired fraction of sevoflurane measured at time to LOER and BIS < or =60 decreases with increase in age. We conclude that inhaled induction with sevoflurane/N2O is dramatically prolonged with increased age. PMID- 16717313 TI - The effect of sevoflurane on ciliary motility in rat cultured tracheal epithelial cells: a comparison with isoflurane and halothane. AB - Halothane and isoflurane potently depress airway ciliary motility. We compared the effect of sevoflurane on ciliary beat frequency (CBF) with that of halothane and isoflurane using purified and cultured rat tracheal epithelial cells. Rat tracheal epithelial cells were isolated from adult male Sprague-Dawley rats to establish an air-liquid interface culture. Apical surfaces of the cells were exposed to a fresh gas containing humidified and warmed (25 degrees C) air (vehicle) with or without sevoflurane (0%-4%), halothane (0%-2%), or isoflurane (0%-2%). The images of motile cilia were videotaped and CBF was analyzed using a computer. Baseline CBF (= 100%) and CBF 30 min after the exposure were measured. CBF 30 min after vehicle exposure was 101% +/- 4% (mean +/- sd). Exposures to 0.25%-2% sevoflurane did not change CBF significantly, although exposures to 0.25%-2% halothane or isoflurane decreased CBF dose-dependently. CBFs 30 min after exposures to 2% of sevoflurane, halothane, and isoflurane were 97% +/- 9%, 56% +/- 14%, and 47% +/- 6%, respectively (n = 5 each). Sevoflurane 4% reduced CBF significantly but slightly (84% +/- 2%, n = 5). These results show that sevoflurane has a direct cilioinhibitory action but its action is much weaker than that of halothane and isoflurane in isolated rat tracheal epithelial cells. PMID- 16717314 TI - The differential effects of halothane and isoflurane on electroencephalographic responses to electrical microstimulation of the reticular formation. AB - Isoflurane and halothane cause electroencephalographic (EEG) depression and neuronal depression in the reticular formation, a site critical to consciousness. We hypothesized that isoflurane, more than halothane, would depress EEG activation elicited by electrical microstimulation of the reticular formation. Rats were anesthetized with either halothane or isoflurane and stimulating electrodes were positioned in the reticular formation. In a crossover design, anesthetic concentration was adjusted to 0.8 and 1.2 minimum alveolar concentration (MAC) of halothane or isoflurane and electrical microstimulation was performed and the EEG responses were recorded. Microstimulation increased the spectral edge and median edge frequencies 2-2.5 Hz at 0.8 MAC for halothane and isoflurane and 1.2 MAC halothane. At 1.2 MAC isoflurane, burst suppression occurred and microstimulation decreased the period of isoelectricity (24% +/- 19% to 8% +/- 7%; P < 0.05), whereas the spectral edge and median edge frequencies were unchanged. At anesthetic concentrations required to produce immobility, the cortex remains responsive to electrical microstimulation of the reticular formation, although the EEG response is depressed in the transition from 0.8 to 1.2 MAC. These data indicate that cortical neurons remain responsive to synaptic input during isoflurane and halothane anesthesia. PMID- 16717315 TI - The effects of xenon on myogenic motor evoked potentials in rabbits: a comparison with propofol and isoflurane. AB - We compared the effects of xenon on myogenic motor evoked potentials (MEPs) with those of propofol and isoflurane in rabbits under ketamine/fentanyl anesthesia. Thirty animals were randomly allocated to one of 3 groups (n = 10 in each group). In the propofol group, propofol was administered at a rate of 0.4 mg x kg(-1) x min(-1) (small) and 0.8 mg x kg(-1) x min(-1) (large). In the isoflurane group, isoflurane was administered at 0.8% (small) and 1.6% (large). In the xenon group, xenon was administered at 35% (small) and 70% (large). Myogenic MEPs in response to stimulation with single pulse and a train of 5 pulses were recorded from the soleus muscle before, during (at small and large doses), and after the administration of each anesthetic. With single-pulse stimulation, MEPs were recorded in 90% and 50% of animals at small and large doses of xenon, respectively, and MEP amplitudes in the xenon and isoflurane groups were significantly lower compared with those in the propofol group. With train pulse stimulation, MEPs were recorded in 100% and 90% of animals at small and large doses of xenon, respectively, and a reduction in MEP amplitudes by xenon was more prominent than by propofol but less than isoflurane at large doses. These results suggest that MEP recording may be feasible under xenon anesthesia if multipulse stimulation is used, although xenon has suppressive effects on myogenic MEPs. PMID- 16717316 TI - The effects of cyclosporin A and insulin on ischemic spinal cord injury in rabbits. AB - We examined the effects of cyclosporin A (CsA), a drug that inhibits mitochondrial permeability transition pore, and insulin on ischemic spinal cord damage in rabbits. We assigned rabbits to 5 groups (n = 6 in each); sham barrier opened group (sham BO), barrier-opened group (BO), barrier-opened-CsA group (BO CsA), barrier-opened-insulin group (BO-I), and barrier-opened-CsA-insulin group (BO-CsA-I). The blood-spinal cord barrier was opened to facilitate drug penetration by a mild injury to the lumber spinal cord on day 1. CsA (10 mg/kg per day IV) was administered on day 3 to day 5 (total 30 mg/kg). Insulin was administered 30 min before ischemia. In all groups, spinal cord ischemia was produced on day 5 by occluding the abdominal aorta for 13 min. Neurological and histopathological evaluations were performed 4 days after ischemia. In group BO CsA, blood glucose concentrations were significantly larger compared with the other four groups, and no protection was observed. In contrast, hindlimb motor function in groups BO-I and Bo-CsA-I and histopathology in group BO-CsA-I were significantly better than in groups sham BO, BO, and BO-CsA. The results indicate that insulin protects against ischemic spinal cord injury, whereas the effect of CsA is, at best, minimal. PMID- 16717317 TI - The neurotoxic effects of amitriptyline are mediated by apoptosis and are effectively blocked by inhibition of caspase activity. AB - Oral tricyclic antidepressants, widely used as adjuncts in the treatment of chronic pain, block sodium channels in vitro and nerve conduction in vivo. However, toxicity of amitriptyline has been observed after neural application. We therefore investigated the mechanism and possible prevention of amitriptyline neurotoxicity. To assess dose-dependent neurotoxicity of amitriptyline, we incubated neuron cultures from adult rat dorsal root ganglia with amitriptyline and quantified neuronal survival. Additionally, we investigated accepted markers of apoptosis (mitochondrial membrane potential, cytosolic cytochrome c, and activated caspase-3) and co-incubated amitriptyline with an inhibitor of caspase activity, z-vad-fmk, to assess the effect on cell survival. We found a dose dependent neurotoxic effect of amitriptyline. Neurons incubated with amitriptyline exhibited loss of mitochondrial membrane potential, release of cytochrome c into the cytoplasm, and activation of caspase-3. Co-incubation with z-vad-fmk substantially improved neuronal survival in culture. In conclusion, amitriptyline-induced neurotoxicity is mediated by apoptosis and is attenuated by inhibition of caspase activity, suggesting that inhibition of apoptotic pathways may be efficient at alleviating local anesthetic-induced neurotoxicity. In vivo studies will have to corroborate whether the co-injection of anti-apoptotic drugs with local anesthetics decreases neurotoxic side effects. PMID- 16717318 TI - A comparison of intracellular lidocaine and bupivacaine concentrations producing nerve conduction block in the giant axon of crayfish in vitro. AB - Clinically, lidocaine requires a larger concentration than bupivacaine to block nerves. Bupivacaine has a higher lipid solubility, tissue permeability, and affinity for sodium channels than lidocaine, resulting in greater anesthetic potency. Local anesthetics require access to the sodium channel from the intracellular milieu. In this study, we sought to determine the intracellular concentration of lidocaine and bupivacaine when a nerve was blocked in the giant axon of a crayfish. A solution of lidocaine or bupivacaine was perfused, and a nerve block was determined as the absence of an evoked action potential after tonic or phasic electrical stimulation. The intracellular lidocaine or bupivacaine concentration was measured using a lidocaine- or bupivacaine sensitive glass micro-electrode. A phasic block was more effectively and rapidly achieved with a smaller concentration of bupivacaine than with lidocaine. The intracellular concentration and intra- to extracellular ratios were significantly larger with lidocaine than with bupivacaine when nerve conduction was blocked. These findings suggest that bupivacaine has a higher potency than lidocaine, at least in the giant axon of a crayfish in vitro. The implications of the present results are that bupivacaine is a more potent nerve block and produces a use dependent (phasic) block at smaller concentrations than lidocaine. PMID- 16717319 TI - Lidocaine inhibition of inducible nitric oxide synthase and cationic amino acid transporter-2 transcription in activated murine macrophages may involve voltage sensitive Na+ channel. AB - Lidocaine has been reported to inhibit nitric oxide (NO) production in activated murine macrophages, but the role of inducible NO synthase (iNOS) in lidocaine induced inhibition of NO has not been explored. In addition, type-2 cationic amino acid transporter (CAT-2) and guanosine triphosphate cyclohydrolase I (GTPCH) also regulate iNOS activity. The effects of lidocaine on CAT-2 and GTPCH are unknown. To explore further these effects, confluent immortalized murine macrophages (RAW264.7 cells) were incubated with lipopolysaccharide (LPS) or in combination with lidocaine (5, 50, or 500 microM) for 18 h before harvesting. We also used tetrodotoxin (TTX) and veratridine to elucidate the possible role of voltage-sensitive Na+ channel. Our data demonstrated that LPS significantly upregulated transcription of iNOS and CAT-2 but not GTPCH in stimulated macrophages. In a dose-dependent manner, lidocaine significantly attenuated the LPS-induced upregulation of iNOS and CAT-2. Conversely, lidocaine significantly increased GTPCH transcription in LPS-stimulated macrophages. The effects of TTX on iNOS, CAT-2, and GTPCH expression were comparable to those of lidocaine. In addition, veratridine significantly attenuated the effects of lidocaine and TTX. We therefore concluded that lidocaine significantly inhibits iNOS and CAT-2 and, in turn, enhances GTPCH transcription in LPS-stimulated macrophages via a mechanism that possibly involves the voltage-sensitive Na+ channel. PMID- 16717320 TI - Electroencephalogram--entropy and acupuncture. AB - Computer-based measuring of the level of sedation and hypnosis is difficult and has proven to be challenging. The electroencephalogram (EEG) has been proposed as a potential method. Response entropy (RE) and state entropy (SE) are multifactor, dimensionless parameters of a new technology of EEG monitoring, and we investigated them for the first time in acupuncture research within this study. Both parameters have been alleged to reflect changes in the clinical state of sedation. Two different acupuncture schemes were tested in a randomized crossover trial with nine healthy volunteers (mean age +/- sd, 28.8 +/- 3.6 yr; 25-36 yr). Applying and stimulating acupuncture needles or performing laserneedle acupuncture at special sedation points decreased RE and SE significantly (P < or = 0.01; paired t-test) compared with the reference interval before acupuncture. In contrast, acupuncture of points for increasing "Qi-energy" did not decrease parameters of entropy. Specific acupuncture schemes produce specific, reproducible, and quantifiable effects on entropy parameters in the EEG. Therefore, entropy measurements during acupuncture seem to be worthy of further evaluation with a larger series of subjects. PMID- 16717321 TI - A new method for estimating arterial occlusion pressure in optimizing pneumatic tourniquet inflation pressure. AB - To reduce pressure-related injuries resulting from pneumatic tourniquet use, the lowest possible inflation pressure is recommended. Arterial occlusion pressure (AOP) is a measure of the cuff pressure required to maintain a bloodless surgical field. However, its determination method is time consuming, requires operator skill, and is therefore seldom used in current practice. An AOP estimation can be made by knowing the pressure transmitted to the underlying soft tissues. We measured upper and lower extremity tissue pressures under the tourniquet cuff at 100, 200, and 300 mm Hg of tourniquet inflation pressures in 30 anesthetized living adult patients. All patients received general anesthesia with neuromuscular relaxation. A Stryker intra-compartmental pressure monitor was used to measure tissue pressures under the tourniquet cuff. In all patients, the soft tissue pressures were consistently lower than the applied tourniquet inflation pressures. Our results revealed tissue padding coefficients for extremities 20 to 75 cm in circumferences. An estimation method of AOP was developed [AOP = (systolic blood pressure + 10)/Tissue padding coefficient]. The new AOP estimation method may be a simple, rapid, and clinically practical alternative to the AOP determination method. PMID- 16717322 TI - The implications of arterial Po2 oscillations for conventional arterial blood gas analysis. AB - In a surfactant-depletion model of lung injury, tidal recruitment of atelectasis and changes in shunt fraction lead to large Pao2 oscillations. We investigated the effect of these oscillations on conventional arterial blood gas (ABG) results using different sampling techniques in ventilated rabbits. In each rabbit, 5 different ventilator settings were studied, 2 before saline lavage injury and 3 after lavage injury. Ventilator settings were altered according to 5 different goals for the amplitude and mean value of brachiocephalic Pao2 oscillations, as guided by a fast responding intraarterial probe. ABG collection was timed to obtain the sample at the peak or trough of the Pao2 oscillations, or over several respiratory cycles. Before lung injury, oscillations were small and sample timing did not influence Pao2. After saline lavage, when Po2 fluctuations measured by the indwelling arterial Po2 probe confirmed tidal recruitment, Pao2 by ABG was significantly higher at peak (295 +/- 130 mm Hg) compared with trough (74 +/- 15 mm Hg) or mean (125 +/- 75 mm Hg). In early, mild lung injury after saline lavage, Pao2 can vary markedly during the respiratory cycle. When atelectasis is recruited with each breath, interpretation of changes in shunt fraction, based on conventional ABG analysis, should account for potentially large respiratory variations in arterial Po2. PMID- 16717323 TI - Paravertebral transcutaneous electrical nerve stimulation reduces movement during general anesthesia with isoflurane. AB - We evaluated paravertebral transcutaneous electrical nerve stimulation (TENS) as a means of enhancing anesthesia during hysterectomy. Patients were randomly assigned to experimental (n = 21) and control (n = 20) groups. Anesthesia with isoflurane was performed uniformly for all patients. Paravertebral (T6 and T7) TENS (50 mA, 15 Hz, continuously) was applied in the experimental group. After 15 min of isoflurane, a lower abdominal, skin-to-adipose-tissue incision was made. Seventeen of 21 patients in the experimental group showed no arm or leg movements during the incision, compared to 8 with 20 patients in the control group (P = 0.007). TENS deserves further exploration as an adjunct technique for general anesthesia. PMID- 16717324 TI - Morphine, oxycodone, methadone and its enantiomers in different models of nociception in the rat. AB - We studied the effects of the commonly used mu-opioid receptor agonists morphine, oxycodone, methadone and the enantiomers of methadone in thermal and mechanical models of acute pain and in the spinal nerve ligation model of neuropathic pain in rats. Subcutaneous administration of morphine, oxycodone, and methadone produced a dose-dependent antinociceptive effect in the tail flick, hotplate, and paw pressure tests. l-methadone, racemic methadone, and oxycodone had a similar dose-dependent antinociceptive effect, whereas the dose-response curve of morphine was shallower. In the spinal nerve ligation model of neuropathic pain, subcutaneous administration of morphine, oxycodone, methadone and l-methadone had antiallodynic effects in tests of mechanical and cold allodynia. l-methadone showed the strongest antiallodynic effect of the tested drugs. d-methadone was inactive in all tests. Morphine 5.0 mg/kg, oxycodone 2.5 mg/kg, and l-methadone 1.25 mg/kg decreased spontaneous locomotion 30 min after drug administration. In conclusion, in acute nociception all mu-opioid receptor agonists produced antinociception, with morphine showing the weakest effect. In nerve injury pain, l-methadone showed the greatest antiallodynic potency in both mechanical and cold allodynia compared with the other opioids. Opioids seem to have different profiles in different pain models. l-methadone should be studied for neuropathic pain in humans. PMID- 16717325 TI - The role of the craniospinal nerves in mediating the antinociceptive effect of transcranial electrostimulation in the rat. AB - Transcranial electrostimulation (TES) has been reported to elicit significant analgesia, but its mechanism of action has not been elucidated. In a recently introduced clinically relevant rat model of TES we have validated and characterized the TES antinociceptive effect, suggesting involvement of the sensory nerves of the rat's scalp in mediating that effect. In this study, we have further investigated the role of the craniospinal nerves by attempting to block the TES antinociceptive effect with local anesthetic injected under the TES electrodes. We also applied different transcutaneous electrical nerve stimulation modalities through the TES electrodes and compared the elicited antinociceptive effect to that of TES. The antinociceptive effect was assessed by measuring nociceptive thresholds in the tail-flick latency test in awake, unrestrained male rats. Data were analyzed by one-way analysis of variance followed by the Bonferroni t-test. The TES antinociceptive effect was significantly reduced after local anesthetic injection, and administration of 100 Hz transcutaneous electrical nerve stimulation was, over time, capable of eliciting the same degree of antinociceptive effect as TES. We conclude that sensory craniospinal nerves play a critical role in mediating the TES antinociceptive action and offer a hypothesis on the underlying mechanism(s) responsible for this action. PMID- 16717326 TI - An evaluation of a polyamine-deficient diet for the treatment of inflammatory pain. AB - Polyamines are thought to be involved in the regulation of numerous metabolic and electrophysiological processes in the nervous system. In this study we evaluated the effect of a synthetic polyamine-deficient diet on pain in a carrageenan (Car) induced inflammatory rat model. Inflammation was induced with a unilateral subcutaneous injection of Car in a plantar hindpaw in rats fed without (control group) or with (deficiency group) a polyamine-deficient diet. Ipsilateral and contralateral hyperalgesia was evaluated using the Randall-Sellito pressure test. Heart rate changes were also recorded under general anesthesia. Then, the effects of a bupivacaine sciatic nerve block and subcutaneous injection of naloxone or ketamine were evaluated for Car-induced hyperalgesia. Data were analyzed using analysis of variance followed by unpaired Student's t-test (significance P < 0.05). Before Car injection, no significant difference was observed in response to mechanical stimuli between the control and the deficiency groups (n = 114 in pooled data). Car injection induced significant ipsilateral and contralateral hyperalgesia in the control groups, whereas a significant analgesic effect appeared in the deficient groups on both the ipsilateral and contralateral hindpaws. This analgesic effect was confirmed by the electrocardiogram recording that showed a significant increase in heart rate in the control group after Car injection compared with the deficiency group that showed a decrease in heart rate under general anesthesia. Bupivacaine sciatic nerve block had no significant effect on hypoalgesia phenomena induced by polyamine deficiency. Naloxone administration had no effect in the control group but reversed the analgesic effect in the deficiency group. Ketamine administration induced a significant analgesic effect in the control group and partly reversed the analgesic effect in the deficiency group. In conclusion, a synthetic polyamine-deficient diet had a significant general analgesic effect on Car-induced mechanical hyperalgesia. The mechanism of analgesic action remains to be elucidated. PMID- 16717327 TI - Morphine-6-glucuronide: morphine's successor for postoperative pain relief? AB - In searching for an analgesic with fewer side effects than morphine, examination of morphine's active metabolite, morphine-6-glucuronide (M6G), suggests that M6G is possibly such a drug. In contrast to morphine, M6G is not metabolized but excreted via the kidneys and exhibits enterohepatic cycling, as it is a substrate for multidrug resistance transporter proteins in the liver and intestines. M6G exhibits a delay in its analgesic effect (blood-effect site equilibration half life 4-8 h), which is partly related to slow passage through the blood-brain barrier and distribution within the brain compartment. In humans, M6G's potency is just half of that of morphine. In clinical studies, M6G is well tolerated and produces adequate and long lasting postoperative analgesia. At analgesic doses, M6G causes similar reduction of the ventilatory response to CO2 as an equianalgesic dose of morphine but significantly less depression of the hypoxic ventilatory response. Preliminary data indicate that M6G is associated less than morphine with nausea and vomiting, causing 50% and 75% less nausea in postoperative and experimental settings, respectively. Although the data from the literature are very promising, we believe that more studies are necessary before we may conclude that M6G is superior to morphine for postoperative analgesia. PMID- 16717328 TI - Failure to recognize loss of incoming data in an anesthesia record-keeping system may have increased medical liability. AB - Automated anesthesia record-keeping systems (AARKs) are increasingly being used. There is a perception that AARKs may limit medical liability. We report a case in which our AARK may have increased our medical liability exposure. Nine months after a patient suffered a serious intraoperative complication, the anesthesiologist was named (as one of several defendants) in a claim alleging failure to properly monitor anesthetic care. One reason why the anesthesiologist was named related to a gap of 93 min in which no vital signs were documented in the anesthesia record. Relying on the physiological monitors to assess the patient's condition, the anesthesiologist did not recognize the interruption of data transmission, because the "active" medication window obscured the graphical display of the vital sign window. PMID- 16717329 TI - Erythromycin promotes gastric emptying during acute pain in volunteers. AB - In this double-blind cross-over study, we assessed whether erythromycin infusion is effective as a prokinetic drug against gastroparesis from acute pain. The effect of erythromycin on gastric emptying (GE) was measured in seven volunteers subjected to a standardized acute painful stimulus. The GE rate for solids was measured using the octanoic acid breath test. An acetaminophen absorption test measured the GE rate for liquids. Five minutes after ingestion of a 13C-labeled meal, the subjects received in randomized order either a test (placebo and erythromycin groups) or a control (control group) stimulus consisting of repeated 1-min immersion of a hand into 4 degrees C (test) or 37 degrees C (control) water, with 15 s for recovery between immersions, for a total of 20 min. While the stimulus was applied, 250 mL saline (control and placebo groups) or 250 mg erythromycin (erythromycin group) was infused. Pain and stress were evaluated using visual analog scales, and standard hemodynamic values were recorded throughout the study. Our results show that acute stress decreased GE for solids, which was significantly accelerated in the erythromycin group in comparison with the placebo group. GE for liquids was similar in the three groups. We conclude that erythromycin is effective as a prokinetic drug for solids in acute painful situations. PMID- 16717330 TI - Plasma level of N terminal pro-brain natriuretic peptide as a prognostic marker in critically ill patients. AB - We studied whether N-terminal pro brain natriuretic peptide (NT-pro BNP) measured at intensive care unit admission is an independent predictor of mortality in critically ill patients. We conducted a prospective observational cohort study enrolling 78 patients with APACHE II scores more than 12. Serum NT-pro BNP and cardiac troponin T were measured at admission, and echocardiography was performed within 24 h. The primary end-point was 30-day mortality. The median NT-pro BNP levels of the 22 (28.2%) patients who died were significantly more frequent than that of those who survived (8328 versus 1016 pg/mL; P = 0.001). Patients with NT pro BNP levels more than 1900 pg/mL had significantly more frequent mortality (47.2% versus 11.9%; P = 0.03). This group also had more frequent moderate to severe left ventricular dysfunction (30.6% versus 9.5%; P = 0.02) and abnormal cardiac troponin T levels (33.3% versus 14.3%; P = 0.05). Multivariate analyses adjusted for APACHE-II revealed that a NT-pro BNP level more than 1900 pg/mL is an independent predictor of mortality. PMID- 16717331 TI - Bolus configuration affects dose requirements of intracarotid propofol for electroencephalographic silence. AB - We hypothesized that an intracarotid bolus injection of propofol to produce electroencephalographic (EEG) silence would require a smaller dose of the drug compared with the continuous infusion of the drug. Furthermore, the bolus propofol dose will be a function of the bolus characteristics in each bolus (mass/volume). We compared the dose requirements of intracarotid propofol needed to maintain EEG silence when delivered as bolus injections to continuous infusions in rabbits. Subsequently, we compared whether four different bolus characteristics (concentration and volume) of propofol (0.33% x 0.1 mL, 0.33% x 0.3 mL, 1% x 0.1 mL, and 1% x 0.3 mL) affected the dose required to produce EEG silence. We found that the infusion rate of propofol required to sustain EEG silence was three-fold larger than the dose required by bolus injections, 22.8 +/ 11.9 vs 6.2 +/- 2.9 mL/h for infusion versus bolus, respectively (n = 7, P < 0.004). Furthermore, during bolus injection, the doses of propofol required to produce EEG silence were a direct function of the bolus volume and the mass of drug in each bolus, total dose = 3.6 + 29 x mg/bolus, n = 32, r = 0.85. For maximum regional effects of the bolus intracarotid drug injection, the bolus characteristics (volume and drug concentration) have to be optimized. PMID- 16717332 TI - The effects of indomethacin on intracranial pressure and cerebral hemodynamics during isoflurane or propofol anesthesia in sheep with intracranial hypertension. AB - The effect of indomethacin in reducing intracranial pressure (ICP) may be dependent on the choice of anesthetic regimen. We studied the effects of indomethacin on ICP and cerebral blood flow (CBF) during isoflurane or propofol anesthesia in a sheep model of intracranial hypertension. A crossover design was applied in which six sheep were anesthetized with isoflurane and propofol in a random order. Anesthetic depth was measured with response and state entropy. Changes in CBF, ICP, mean arterial blood pressure, arterio-venous oxygen difference, and Paco2 were measured at specific times before and after an IV indomethacin bolus (0.2 mg/kg). Response and state entropy values during anesthesia were similar in both groups. Isoflurane and propofol reduced CBF by 11% and 34%, respectively. Indomethacin caused a reduction in ICP within 15 s during both anesthetic regimens, with the decrease in ICP being significantly more pronounced during isoflurane (P = 0.009). In both anesthetic groups, indomethacin caused a simultaneous increase in mean arterial blood pressure and a further 17% versus 14% decrease in CBF from predrug values for isoflurane and propofol, respectively. The reduction in CBF was significantly more pronounced for propofol (P = 0.02). The effect on ICP, however, was most pronounced during isoflurane anesthesia. We suggest that the effect of indomethacin is partly mediated by an autoregulatory response. PMID- 16717333 TI - The effect of desflurane and sevoflurane on cerebral oximetry under steady-state conditions. AB - We studied the effect of sevoflurane and desflurane on regional cerebral oxygenation (rSO2). Twenty-two patients undergoing abdominal hysterectomy received sevoflurane and desflurane for 15 min each and 30 min apart under steady state conditions in a randomized, crossover manner to maintain a bispectral index (BIS) of 40-50. In another 22 patients undergoing the same anesthesia and surgery BIS was maintained at 20-30. During the 15-min administration of each anesthetic at steady-state conditions rSO2, BIS, inspired and end-tidal anesthetic concentrations, end-tidal CO2, Spo2, systolic and diastolic blood pressures, and heart rate were recorded every 3 min. The rSO2 did not differ between sevoflurane and desflurane when BIS values were maintained between 40-50 or 20-30. The MAC(BIS) values required to maintain BIS at 40-50 and at 20-30 were 1.0 versus 1.2 (P = 0.004) and 1.6 versus 1.8 (P < 0.001) for desflurane and sevoflurane respectively. Higher rSO2 values were obtained by 1.6 MAC (71 +/- 13) than by 1 MAC of desflurane (66 +/- 10; P < 0.001) and by 1.8 MAC (72 +/- 11) than by 1.2 MAC of sevoflurane (66 +/- 13; P < 0.001). In conclusion, equipotent concentrations of desflurane or sevoflurane in terms of BIS are associated with similar rSO2 values, but larger anesthetic concentrations of both anesthetics increased the rSO2 values. PMID- 16717334 TI - The use of hypertonic saline for treating intracranial hypertension after traumatic brain injury. AB - The past decade has witnessed a resurgence of interest in the use of hypertonic saline for low-volume resuscitation after trauma. Preliminary studies suggested that benefits are limited to a subgroup of trauma patients with brain injury, but a recent study of prehospital administration of hypertonic saline to patients with traumatic brain injury failed to confirm a benefit. Animal and human studies have demonstrated that hypertonic saline has clinically desirable physiological effects on cerebral blood flow, intracranial pressure, and inflammatory responses in models of neurotrauma. There are few clinical studies in traumatic brain injury with patient survival as an end point. In this review, we examined the experimental and clinical knowledge of hypertonic saline as an osmotherapeutic agent in neurotrauma. PMID- 16717335 TI - The effects of age on the median effective concentration of ropivacaine for motor blockade after epidural anesthesia with ropivacaine. AB - Minimal local analgesic concentrations have been defined as the median effective concentration (EC50). In this study, we sought to examine the effect of age on motor blockade and determine the motor block EC50 of elderly patients after epidural administration of ropivacaine in patients undergoing urological or minor lower limb surgery. ASA physical status I-II patients were enrolled in 1 of 2 age groups (Group 1: > or =70 yr; Group 2: <70 yr). Each received a 15-mL bolus of epidural ropivacaine without epinephrine. The first patient in each group received 0.425%. Up-down sequential allocation was used to determine subsequent concentrations at a testing interval of 0.025%. Effective motor blockade was defined as a modified Bromage score >0 within 30 min. The motor blockade EC50 of ropivacaine was 0.383% (95% confidence interval, 0.358%-0. 409%) in group 1 and 0.536% (95% confidence interval, 0.512%-0.556%) in group 2 (P < 0.01). We conclude that age is a determinant of motor blockade EC50 of ropivacaine with epidural administration. PMID- 16717336 TI - A comparison of epidural blockade produced by plain 1% lidocaine and 1% lidocaine prepared by dilution of 2% lidocaine with the same volume of saline. AB - Local anesthetics are commonly diluted with saline, but the influence of the dilution on the epidural anesthesia remains unclear. We randomized 40 patients scheduled for gynecological abdominal surgery under epidural anesthesia to one of two groups; those in group P received plain commercially prepared 1% lidocaine and those in group D received 1% lidocaine derived from 2% lidocaine and the same volume of saline was infused epidurally with an epidural catheter at L1-2. The pH and sodium and chloride ion concentrations of the solutions were measured. Sensory and motor blockade, foot skin temperature, arterial blood pressure, and heart rate were assessed at 5, 10, and 15 min after the epidural infusion. The spread of sensory blockade was significantly wider in group P at all assessment times than in group D. The increase of foot temperature and decrease of mean arterial blood pressure were significantly faster in group P than in group D. Although the mean pH values of the two solutions were similar, sodium and chloride ion concentrations of the diluted solution were significantly larger than those of the plain solution. We conclude that 2% lidocaine diluted with the same volume of saline produces less potent epidural blockade than commercially prepared plain 1% lidocaine. PMID- 16717337 TI - A comparison between double-injection axillary brachial plexus block and midhumeral block for emergency upper limb surgery. AB - In this prospective and randomized study, we compared a double-injection axillary (median and radial nerves) block with a midhumeral block in 90 patients undergoing emergency upper limb surgery. Time to perform the block, success rate, and patient tolerance were evaluated. The time to perform the block was 5 min longer in the midhumeral group. The success rate was similar in both groups (80% and 91% in groups axillary and midhumeral respectively), except for the musculocutaneous nerve. Patient tolerance was better in the axillary group. Double-injection axillary brachial plexus block is superior to midhumeral block for emergency hand surgery. PMID- 16717338 TI - The femoral nerve and its relationship to the lateral circumflex femoral artery. AB - Small branches of the femoral artery in the femoral triangle are not palpable and could increase the risk of intravascular injection during femoral nerve (FN) block. I evaluated the position of the lateral circumflex femoral artery (LCFA), a lateral branch of the femoral artery, in relationship to accepted landmarks for FN blockade, including the inguinal ligament, inguinal crease, and FA. Forty cadaver lower extremities were dissected. In 50% of specimens, the LCFA, as it crossed the FN, was within 1 cm of the inguinal crease, the recommended level for needle insertion for FN blockade. The mean depth of this artery at the inguinal crease was 1.7 cm, whereas the mean depth of the FN was 1.1 cm at this level. In most specimens, the LCFA coursed between the branches of the FN, although it sometimes lay deep to all of the branches. Knowledge of this anatomy may allow for safer FN blockade. PMID- 16717339 TI - Cauda equina syndrome and profound hearing loss after spinal anesthesia with isobaric bupivacaine. AB - A 33-yr-old man undergoing anorectal surgery developed cauda equina syndrome and bilateral profound hearing loss after single-injection spinal anesthesia with isobaric bupivacaine. There was no pain on needle placement. Neurologic assessment found impaired sensation to pinprick in the perineal region, lower extremity paralysis, and bowel and bladder incontinence. In addition, he developed a bilateral profound hearing loss involving the low frequencies, with the left side more affected than the right side. Although hearing impairment can occur with cerebrospinal fluid leakage, the etiology of cauda equina syndrome is uncertain. The simultaneous occurrence of these events has not been previously reported. PMID- 16717340 TI - Divergence paresis without positional headache: an unusual presentation of cerebrospinal fluid hypovolemia after spinal anesthesia. AB - We report a rare complication of spinal anesthesia-divergence paresis-which is characterized by an acquired horizontal diplopia at distance without evidence for abducens palsy. A 64-yr-old man underwent prostatectomy under spinal anesthesia with 2.5 mL of dibucaine hydrochloride 0.3% injected through a 20-gauge cutting tip spinal needle. Seventeen days after the operation, the patient noticed horizontal diplopia for distant objects. Although cranial magnetic resonance imaging demonstrated diffuse pachymeningeal gadolinium enhancement and subdural effusion, characteristic findings of cerebrospinal fluid hypovolemia, the patient had no positional headache. Gadolinium-enhanced magnetic resonance imaging may be useful when a patient develops neurologic symptoms after dural puncture. PMID- 16717341 TI - A systematic review (meta-analysis) of the accuracy of the Mallampati tests to predict the difficult airway. AB - The original and modified Mallampati tests are commonly used to predict the difficult airway, but there is controversy regarding their accuracy. We searched MEDLINE and other databases for prospective studies of patients undergoing general anesthesia in which the results of a preoperative Mallampati test were compared with the subsequent rate of difficult airway (difficult laryngoscopy, difficult intubation, or difficult ventilation as reference tests). Forty-two studies enrolling 34,513 patients were included. The definitions of the reference tests varied widely. For predicting difficult laryngoscopy, both versions of the Mallampati test had good accuracy (area under the summary receiver operating characteristic (sROC) curve = 0.89 +/- 0.05 and 0.78 +/- 0.05, respectively). For predicting difficult intubation, the modified Mallampati test had good accuracy (area under the sROC curve = 0.83 +/- 0.03) whereas the original Mallampati test was poor (area under the sROC curve = 0.58 +/- 0.12). The Mallampati tests were poor at identifying difficult mask ventilation. Publication bias was not detected. Used alone, the Mallampati tests have limited accuracy for predicting the difficult airway and thus are not useful screening tests. PMID- 16717342 TI - Body position affects manual dexterity. AB - To evaluate the relationship between posture and psychomotor efficiency, 20 anesthesia providers performed a test requiring manual dexterity and eye-hand coordination while seated, kneeling, and standing bent forward at the waist. Performance on the Grooved Pegboard Test improved 6%-10% (P < 0.005) when subjects were seated compared to kneeling and standing bent at the waist, positions that the subjects rated as more painful and less comfortable than sitting. Sitting improved manual dexterity more in subjects with poor scores than it did in subjects with good performance. This study demonstrates improved manual dexterity in subjects seated in a comfortable position. PMID- 16717343 TI - Risk factors for postoperative nausea and vomiting. AB - Knowledge of postoperative nausea and vomiting (PONV) risk factors allows anesthesiologists to optimize the use of prophylactic regimens. Modern PONV risk research began in the 1990s with publication of studies using logistic regression analysis to simultaneously identify multiple independent PONV predictors and publication of meta-analyses and systematic reviews. This literature shows that female gender post-puberty, nonsmoking status, history of PONV or motion sickness, childhood after infancy and younger adulthood, increasing duration of surgery, and use of volatile anesthetics, nitrous oxide, large-dose neostigmine, or intraoperative or postoperative opioids are well established PONV risk factors. Possible risk factors include history of migraine, history of PONV or motion sickness in a child's parent or sibling, better ASA physical status, intense preoperative anxiety, certain ethnicities or surgery types, decreased perioperative fluids, crystalloid versus colloid administration, increasing duration of anesthesia, general versus regional anesthesia or sedation, balanced versus total IV anesthesia, and use of longer-acting versus shorter-acting opioids. Early-phase menstruation, obesity and lack of supplemental oxygen are disproved risk factors. Current risk scoring systems have approximately 55%-80% accuracy in predicting which patient groups will suffer PONV. Further research examining genetic and under-investigated clinical patient characteristics as potential risk factors, and involving outpatients and children, should improve predictive systems. PMID- 16717344 TI - Pulmonary artery catheter burn after multiple transthoracic defibrillatory shocks. PMID- 16717345 TI - Retrieval of aspirated endotracheal tube. PMID- 16717346 TI - Paraplegia after spinal anesthesia as a result of previously undiagnosed vertebral tuberculosis. PMID- 16717347 TI - Circumferential subconjunctival anesthesia versus retrobulbar anesthesia for extracapsular cataract extraction. PMID- 16717348 TI - Emergent facial lacerations repair in children: nerve blocks to the rescue! PMID- 16717349 TI - Patient-controlled interscalene analgesia after shoulder surgery: catheter insertion by the posterior approach. PMID- 16717351 TI - Hypercarbia and arrhythmias resulting from faulty Bain circuit: a report of two cases. PMID- 16717350 TI - Activated recombinant factor VII to reverse oral anticoagulants for emergent cesarean delivery. PMID- 16717352 TI - Initial distribution volume of glucose early after cardiac surgery. PMID- 16717353 TI - Immediate allergic reaction to betamethasone during anesthesia. PMID- 16717354 TI - Can conventional peripheral nerve stimulators induce direct muscle depolarization? PMID- 16717355 TI - Frenular injury during insertion of the ProSeal laryngeal mask airway using the introducer tool technique. PMID- 16717356 TI - An unusual capnogram in a pediatric patient. PMID- 16717357 TI - Unintended consequences? Unanswered questions? PMID- 16717358 TI - Bolus vasopressin during hemorrhagic shock? PMID- 16717359 TI - Subarachnoid anesthesia for cesarean delivery in a parturient with Strumpell Lorrain disease. PMID- 16717360 TI - From the anesthesiologist's perspective: placement of vagal nerve stimulator. PMID- 16717361 TI - Stimulating cervical epidural catheter. PMID- 16717362 TI - Neck contracture release and reinforced tracheal tube obstruction. PMID- 16717363 TI - Rocuronium versus succinylcholine for rapid tracheal intubation. PMID- 16717364 TI - Time to accept allopathic physicians into AOA-approved residencies? PMID- 16717365 TI - Will the last DO turn off the lights? PMID- 16717366 TI - The national report card on the state of emergency medicine. PMID- 16717367 TI - Mechanism-of-injury approach to evaluating patients with blast-related polytrauma. AB - Civilians and military personnel alike are increasingly being exposed to explosives in war zones and other regions of political conflict and, consequently, they are suffering associated blast-related polytrauma (multiple complex injuries). Although acute, emergency-based medical care for patients with blast-related trauma has been well described, postacute clinical management- which is of greatest interest to primary care physicians and rehabilitation specialists--has not been well discussed or researched. The authors offer a description of the common injuries seen in patients with blast-related polytrauma, as well as a conceptual model of a potential evaluation and treatment strategy in the postacute setting. Although medical evaluation of a patient typically proceeds in a sequential manner based on primary symptoms, the authors advocate a parallel approach to patient evaluation based on mechanism (cause) of injury. Such an approach relies on knowledge of the typical physical and psychological sequelae associated with a particular mechanism of injury to guide patient assessment and treatment. The authors highlight the mechanism-of-injury approach used with patients who have blast-related polytrauma at the Veterans Health Administration's (VHA) Veterans Affairs Medical Center in Tampa, Fla, site of one of the VHA's four Polytrauma Rehabilitation Centers. PMID- 16717368 TI - Nonobstetric lacerations of the vagina. AB - Although obstetrically related trauma remains the most common cause of injury to the female genital tract, trauma of nonobstetric origin is not uncommon. Reports of traumatic injuries to the vagina, especially lacerations, have been infrequent in the literature and offer only a generalized approach to this problem. Severe vaginal lacerations may result in life-threatening blood loss. The authors report their recent experience with treating patients who have this type of trauma, review mechanisms of injury, and provide an organized treatment protocol for the nonobstetric patient with suspected vaginal laceration. Preparation for these emergencies circumvents dangerous delays and inadequate examination and treatment. PMID- 16717369 TI - Characteristics of osteopathic physicians choosing to practice rural primary care. AB - As increasing numbers of women and osteopathic physicians enter medical practice, it is important to identify their choices in medical specialty and practice location and the implications these choices have for the future of healthcare in the United States. In 2003, data from the Texas Medical Board and the Office of the [Texas] State Demographer were aggregated to compare the rates at which physicians differed in their choices to practice primary care specialties in a rural location. In addition, the impact of sex and type of medical degree on these choices was examined. Analyses revealed that male osteopathic physicians were 2.3 times more likely than all other physician groups to practice rural primary care. Analyses also revealed that female osteopathic physicians were more likely than other physicians to choose primary care as a specialty and were 2.5 times more likely than female allopathic physicians to practice primary care in a rural location. Policies intended to produce primary care or rural primary care physicians should take into account the effects of gender and osteopathic training. PMID- 16717370 TI - A 44-year-old woman with hematemesis and cutaneous hemorrhages as a result of superwarfarin poisoning. AB - The authors present the case of a 44-year-old American Indian woman with hematemesis, spontaneous cutaneous hemorrhages, and multiple ecchymoses. Coagulation factor analyses demonstrated both prolonged prothrombin time (PT, >40 s) and prolonged partial thromboplastin time (PTT, >120 s). Measurement of the serum level of brodifacoum (37 ng/mL), one of the superwarfarin agents commonly used in rodenticides, confirmed poisoning as the cause of the patient's symptoms. Substantial amounts of fresh frozen plasma and vitamin K were required to obtain normal coagulation parameters and maintain these parameters over a 3-week inhospital period. Oral administration of vitamin K (100 mg daily) maintained normal PT (14.1 s), PTT (33.0 s), and international normalized ratio (INR, 1.48) at 2 weeks after the patient was discharged from the hospital. By 2 months postdischarge, PT, PTT, and INR returned to elevated levels because of patient noncompliance with the prescribed tapering vitamin K regimen. PMID- 16717371 TI - Osteopathic manipulative treatment of a 26-year-old woman with Bell's palsy. AB - Bell's palsy is caused by a lesion of the facial nerve and results in unilateral paralysis or paresis of the face. The condition affects approximately 23 in 100,000 persons, with onset typically occurring between the ages of 10 and 40 years. The authors report the case of a 26-year-old woman with Bell's palsy, whom they treated with osteopathic manipulative treatment that was focused on the enhancement of lymphatic circulation. The osteopathic manipulative procedures used involved reducing restrictions around four key diaphragms (thoracic outlet, respiratory diaphragm, suboccipital diaphragm, cerebellar tentorium), as well as applying the thoracic pump, muscle energy, primary respiratory mechanism, and osteopathy in the cranial field. The authors, who were guided by the four principles of osteopathic philosophy, report that the patient's symptoms resolved within 2 weeks, during which two sessions of osteopathic manipulative treatment, each lasting approximately 20 minutes, were held. Patient recovery occurred without the use of pharmaceuticals. PMID- 16717372 TI - Relationships between scores on the COMLEX-USA Level 2-Performance Evaluation and selected school-based performance measures. AB - The Comprehensive Osteopathic Medical Licensing Examination USA Level 2 Performance Evaluation (COMLEX-USA Level 2-PE) is a national multistation performance examination designed to examine students' osteopathic clinical skills. The current study examines the relationship between achievement levels on the COMLEX-USA Level 2-PE and selected school-related variables for the class of 2005 at the West Virginia School of Osteopathic Medicine in Lewisburg, WVa (N=70). Significant (P<.01) correlations between the COMLEX-USA Level 2-PE summary performance and selected academic achievement measures include: weighted Physical Diagnosis grade, 0.41; weighted year 1 and year 2 Osteopathic Principles and Practice grade, 0.37: overall year 2 grade point average, 0.42; the objective structured clinical evaluation (OSCE) Physical Examination score, 0.40; and the OSCE Total Station score, 0.33. While further research is needed, the current study found modest but notable relationships between school-generated academic variables and performance on the COMLEX-USA Level 2-PE, and therefore supports the validity of the COMLEX-USA Level 2-PE examination for assessing the clinical skills of future osteopathic physicians. PMID- 16717373 TI - Survey on the clinical skills of osteopathic medical students. AB - As part of the standard-setting methods used by the National Board of Osteopathic Medical Examiners for its Comprehensive Osteopathic Medical Licensing Examination clinical skills performance evaluation (COMLEX-USA Level 2-PE), a self administered survey was distributed electronically and by mail to deans of colleges of osteopathic medicine, directors of graduate medical education programs, osteopathic medical students, and experts chosen demographically to represent osteopathic physicians in the United States. Groups were asked to rate fourth-year osteopathic medical students and interns on their clinical skills and acceptable pass rates and expected pass rates on the COMLEX-USA Level 2-PE. The surveys were not used systematically to compute the passing standards but to provide additional support for their validity. The viewpoints of the deans differed from those of the students, osteopathic graduate medical education program directors, and experts regarding clinical skills proficiencies and acceptable pass rates. However, all of the groups agreed that, on average, some students and interns do not have adequate clinical skills. These results provide additional support for requiring acceptable performance on a comprehensive clinical skills examination before admission to osteopathic graduate medical education programs. PMID- 16717375 TI - Progressive idea for osteopathic medical education. PMID- 16717378 TI - Osteopathic medical research. PMID- 16717376 TI - Manual therapies. PMID- 16717379 TI - Osteopathic manipulative treatment is for all DOs. PMID- 16717380 TI - Osteopathic physicians and disaster relief. PMID- 16717381 TI - Welcoming new graduates: some lessons learned. PMID- 16717382 TI - Anticoagulant and antiplatelet medications. PMID- 16717384 TI - Africanized killer bees: a case study. PMID- 16717385 TI - Amiodarone-induced thyroid dysfunction. PMID- 16717387 TI - Teaching cardiopulmonary resuscitation via the Web. PMID- 16717386 TI - Development and implementation of a multidisciplinary sepsis protocol. PMID- 16717388 TI - Congenital tracheal stenosis. PMID- 16717389 TI - Ask the experts. The standard of care for cardiac surgery recovery in the area of weaning. PMID- 16717391 TI - RN-driven, evidence-based practice. PMID- 16717392 TI - Facile syntheses of acyl dihydroxyacetone phosphates and lysophosphatidic acids having different acyl groups. AB - In this study, we report novel and simple chemical syntheses of acyl dihydroxyacetone phosphate (DHAP) and 1-acyl glycero-3-phosphate [lysophosphatidic acid (LPA)], key intermediaries in the formation of glycerolipids containing ester and ether bonds. The synthesis of acyl DHAPs involved acylating the dimethyl ketal of DHAP by acid anhydride using 4 pyrrolidinopyridine as the catalyst, and the resulting product was deketalized by HClO(4) in acetone to produce acyl DHAP. The acid anhydride was either added directly or generated in the reaction mixture from the corresponding fatty acid using dicyclohexylcarbodiimide as the condensing agent. Using these methods, a number of acyl DHAPs having short-, medium-, and long-chain saturated and unsaturated acyl groups were synthesized, with overall yields from 37% to 75%. The activities of these acyl DHAPs as substrates for guinea pig liver peroxisomal acyl DHAP:NADPH reductase and alkyl DHAP synthase were then determined. Next, starting from these acyl DHAPs, a variety of LPAs were synthesized by chemical reduction of the ketone group. Biological activities of these LPAs were determined by measuring their relative abilities to release intracellular Ca(2+) via the LPA receptor. A combined chemical-enzymatic method is also described to prepare the natural LPA from the racemic mixture. PMID- 16717394 TI - A case of hepatitis C-associated osteosclerosis in an elderly Japanese man. AB - Hepatitis C-Associated Osteosclerosis (HCAO) is characterized by a marked increase in bone mass with deep bone pain. Since 1992, eleven cases of HCAO have been reported. This report describes an elderly Japanese man with HCAO, whose clinical course we followed for 3 years. A 68-year-old man developed pain in both pretibial regions in June 2000, and he had frequent episodic loss of muscular strength in his hands. He had recieved blood transfusion for a bleeding ulcer 43 years before and was seropositive for hepatitis C virus. His serum alkaline phosphatase (ALP) level was markedly increased, while his serum calcium was slightly decreased and serum phosphate was normal. Skeletal radiographs of the lower extremities showed a progressive increase in skeletal density, but did not show any apparent deformity. Administration of nonsteroidal anti-inflammatory drugs led to a reduction in bone pain. Treatment with vitamin D3 and calcium decreased the number of episodes of sudden muscular weakness and maintained serum calcium within the normal range. Three years after the onset of the disease, bone mineral density of his lumbar vertebrae and left hip rose from 0.963 g/cm2 to 1.096 g/cm2, and from 0.938 g/cm2 to 1.383 g/cm2, respectively. His serum ALP level decreased from 2889 to 277 IU/L (normal range: 104-338) and serum calcium normalized. These findings were accompanied by a decrease in bone pain. This case and previous reports suggest that the skeletal tissue of this disease appears to be of good quality. PMID- 16717395 TI - Body mass index negatively influences glycated albumin, but not glycated hemoglobin, in diabetic patients. AB - Measurement of serum glycated albumin (GA) is accepted as an alternative method to evaluate chronic glycemic control in diabetic patients in whom measurement of HbA 1c is inadequate for some reason. Although GA levels are known to be influenced by serum albumin turnover besides glycemia, little is known about the physiological and pathological conditions affecting GA levels. This study was aimed to prove the effects of body mass index (BMI) on GA measurement in diabetic patients. We studied 209 patients with type 2 diabetes mellitus whose HbA 1c levels had been stable for at least the past three months. In the study patients HbA 1c and GA levels were found to be correlated to one another. Fasting plasma glucose (FPG) was significantly correlated with HbA 1c and GA. BMI showed a significant negative correlation with GA levels, whereas there was no correlation of BMI with HbA 1c levels. Multivariate regression analyses revealed that only FPG was positively correlated with HbA 1c, while FPG was positively and BMI was negatively correlated with GA. Only BMI was negatively correlated with the ratio of GA to HbA 1c. These results clearly demonstrate that GA levels are negatively influenced by BMI in diabetic patients. PMID- 16717396 TI - Profound reduction in T-helper (Th) 1 lymphocytes in peripheral blood from patients with concurrent type 1 diabetes and Graves' disease. AB - Type 1 diabetes likely is mediated by T-helper (Th) 1 lymphocytes, while Graves' disease may involve Th2 predominance. We investigated the balance between Th1 and Th2 cells and between Th1- and Th2-associated chemokine receptor expression on peripheral lymphocytes in subjects including patients with coexisting type 1 diabetes and Graves' disease. Peripheral blood mononuclear cells of all subjects were examined by flow cytometry for intracellular cytokines (IFN-gamma for Th1; IL-4 for Th2) and expression of the chemokine receptors CXCR3 (Th1-associated) and CCR4 (Th2-associated). Plasma concentrations of interferon-inducible protein (IP)-10, a CXCR3 ligand, and thymus and activation-regulated chemokine (TARC), a CCR4 ligand, were measured by enzyme-linked immunosorbent assays. IFN-gamma producing-T lymphocytes were significantly fewer in patients with coexisting type 1 diabetes and Graves' disease (12.4 +/- 6.8%, n = 6) than in healthy control subjects (19.9 +/- 4.1%, n = 6; P < 0.01) or patients with type 2 diabetes (19.1 +/- 4.5%, n = 5; P < 0.05). We found no significant difference in IFN-gamma producing T lymphocytes between healthy controls and patients with only type 1 diabetes (n = 8) or Graves' disease (n = 5). Plasma IP-10 concentrations were significantly higher in patients with coexisting type 1 diabetes and Graves' disease than in control subjects (106.3 +/- 30.48 vs. 66.7 +/- 25.3 pg/ml, P = 0.0343). Considering only patients with type 1 diabetes alone, duration of diabetes correlated positively with IFN-gamma-producing T lymphocytes (r = 0.773, P = 0.0242) and the ratio of CXCR3 to CCR4 receptor expression (r = 0.947, P = 0.0004). In conclusion, Th1-associated T lymphocytes were fewer in peripheral blood from patients having both type 1 diabetes and Graves' disease than in those with either disease alone. Numbers of peripheral Th1 lymphocytes increased with increasing time from onset of type 1 diabetes in patients with type 1 diabetes alone. PMID- 16717397 TI - Lack of puberty despite elevated estradiol in a 46,XY phenotypic female with Frasier syndrome. AB - Frasier syndrome is characterized by slowly progressive nephropathy, male pseudohermaphroditism, streak gonad, and high risk of gonadoblastoma development. Here we report a case of a 46,XY phenotypic female with Frasier syndrome, who was under hemodialysis. While her serum estradiol level was gradually increasing annually, gonadotropin level was constantly extremely high, and her appearance was still prepubertal. She was heterozygous for a novel guanine>adenine point mutation at position +1 of the splice donor site within intron 9 (IVS 9 + 1G>A) of the Wilms' tumor 1 gene. The possibility of this disease should be taken into consideration whenever we encounter a patient with steroid-resistant nephrotic syndrome and delayed puberty. PMID- 16717398 TI - A case of sleep apnea syndrome manifesting severe hypertension with high plasma norepinephrine levels. AB - A 55-year-old female was admitted to our hospital with severe hypertension (274/140 mmHg). Endocrinological examination revealed that her plasma levels of norepinephrine (NE) was elevated with high levels of urinary NE, normetanephrine and vanillylmandelic acid (VMA), suggesting the presence of pheochromocytoma. However, neither computed tomography nor MIBG scintigraphy detected any catecholamine-producing tumor in or outside the adrenal glands. She was screened with full polysomnography because of heavy snoring, and the diagnosis of severe obstructive sleep apnea syndrome (OSAS) was made. She was treated with calcium channel blocker for three weeks, but severe hypertension persisted. After treatment with nasal continuous positive airway pressure (CPAP) was added, her blood pressure gradually lowered week by week. Concomitantly, the levels of plasma and urinary NE, urinary normetanephrine and urinary VMA were normalized following nasal CPAP therapy for 2 weeks. Additional treatments with alpha adrenergic blocker further decreased her home blood pressure. After a year, she continued nasal CPAP therapy and her blood pressure was nearly below 160/100 mmHg. Urinary NE level was slightly above normal range and other catecholamines stayed within the normal range. This case shows that patients with OSAS could develop severe hypertension through elevated sympathetic tone, mimicking pheochromocytoma. Nasal CPAP therapy is recommended not only to improve hypertension and catecholamine excess but also to distinguish the condition from pheochromocytoma. PMID- 16717399 TI - Effects of erythropoietin on cardiac remodeling after myocardial infarction. AB - Erythropoietin (EPO) has been suggested to have a cardioprotective effect against ischemia. The purpose of this study was to examine the effects of EPO on cardiac remodeling after myocardial infarction (MI). MI was induced by ligation of the coronary artery in Wistar rats. The rats with MI were randomly divided into untreated MI and two EPO-treated MI groups. EPO was administered subcutaneously by injection once a day for 4 days after MI at 5000 U/kg or 3 times a week for 4 weeks at 1000 U/kg. Five days after MI, EPO prevented the increase in activated caspase 3, matrix metalloproteinase-2, and transcriptional activation of activator protein-1 in non-infarcted myocardium. Four weeks after MI, left ventricular weight, left ventricular end-diastolic pressure, and left ventricular dimension were increased, and ejection fraction and E wave deceleration time were decreased. EPO significantly attenuated this ventricular remodeling and systolic and diastolic dysfunction. In addition, EPO significantly attenuated the interstitial fibrosis and remodeling-related gene expression in non-infarcted myocardium. Furthermore, EPO significantly enhanced angiogenesis and reduced apoptotic cell death in peri-infarcted myocardium. In conclusion, when administered after MI, EPO prevents cardiac remodeling and improves ventricular function with enhanced angiogenesis and reduced apoptosis. PMID- 16717401 TI - Platelet separation from whole blood in an aqueous two-phase system with water soluble polymers. AB - Platelet-rich plasma (PRP) stimulates tissue healing and centrifugation is the only method for PRP preparation. The purpose of the present study was to develop a method to separate platelets without centrifugation. We used 16 polymers of different chemical characteristics and mixed each polymer with fresh whole blood containing anticoagulant citrate-dextrose. Then, we observed blood cell separation. Focusing on the effective polymers and comparing our method with the conventional centrifugation method, we examined platelet recovery rate and P selectin expression, which represents platelet activation. Poly-L-glutamic acid (PGA) and poly-L-aspartic acid (PAA) separated platelets efficiently; however, these polymers activated platelets. On the contrary, poly (2-methacryloyloxyethyl phosphorylcholine-co-n-butyl methacrylate) (PMB), which has phosphorylcholine residues mimicking a cellular membrane molecule, separated platelets moderately and the PMB-separated platelets were nearly inactive compared to the separation with the ordinary centrifugation and PGA. Conclusively, the present experiments demonstrated that without centrifugation platelets can be separated from whole blood with some water-soluble polymers, such as PGA, PAA, and PMB, and that PMB has an advantage not to activate platelets. PMID- 16717400 TI - Screening for control genes in mouse hippocampus after transient forebrain ischemia using high-density oligonucleotide array. AB - In conventional relative gene expression analysis (Northern blotting, RT-PCR, and in situ hybridization), housekeeping genes such as the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and beta-actin genes, whose expression levels are considered stable, have been used as control genes for normalization of RNA quantitation. However, it has been reported that the expression levels of these two control genes are affected by ischemia. Therefore, we have been searching for novel control genes whose expression levels are stable in a mouse model of transient forebrain ischemia. Using the GeneChip Mu6500 array set, we monitored the expression levels of approximately 6000 murine genes in the mouse hippocampus during 24 h of ischemia-reperfusion. To select stable genes, we applied the restricted criterion of a 1.5-fold change in expression level as the threshold. By adding statistical analysis with this criterion, we identified 10 genes as candidates for control genes from the GeneChip data. In this criterion, GAPDH and beta-actin genes were not included in the 10 genes as candidates for control genes. The present findings might be relevant to the use of control genes in quantitation of RNA, particularly in the study of mouse transient forebrain ischemia. PMID- 16717402 TI - Supplementation with urea and molasses and body weight, milk yield and onset of ovarian cyclicity in cows. AB - Ten multiparous crossbred local Zebu cows were randomly divided into two nutritional groups (A and B) to determine the effect of urea-molasses-mineral block supplementation on body weight gain, milk production, and onset of ovarian cyclicity after calving. Both groups had farm rations daily, but the supplemented group (B) was provided with an additional diet daily of 250 g urea-molasses mineral block. The cows in group A required 80-120 days (98.0 +/- 6.7 days) until peak milk progesterone concentrations and 60-80 days (72.0 +/- 3.8 days) were required for group B (p<0.05). Group B needed a shorter period for expression of standing estrus (91-101 days; mean 96.2 +/- 2.3 days) than group A [130-153 (141.6 +/- 4.6) days; p<0.01]. For groups A and B, body weight gain was 8.4 +/- 3.4 kg and 18.4 +/- 3.2 kg, respectively (p<0.01). The average milk production of groups A and B were 3.3 +/- 1.0 and 4.8 +/- 1.6 L/day, respectively (p>0.05). There was linear improvement in milk yield from Day 60 postpartum up to Day 90 of lactation in group B (supplemented). However, in group A, milk production decreased starting on Day 40 after parturition. PMID- 16717403 TI - [Equipment at the special nursing homes for the elderly: a workplace survey of new nursing homes in Osaka Prefecture]. AB - The number of those who need nursing care and the workers who provide it have been increasing year after year. In April 2000, the public nursing care insurance system was enacted by the Japanese government. After its enaction, care equipment came under scrutiny, but the situation regarding the installation of equipment is not fully understood. In order to understand the present state of care equipment in nursing facilities for the elderly, we conducted a workplace and interview survey. The surveyed facilities were 10 special nursing homes for the elderly in Osaka Prefecture which were established after April 2002. The average number of elderly residents was 79.0, the average value of degree of care was 3.52, and the average number of caregivers was 28.3 per facility. We found all facilities had installed some kinds of bathing equipment: stretcher type, 9 facilities; bath chair type, 8 facilities. In the facilities with bath-chairs, 6 facilities had special bathtubs, and 6 facilities had general bathtubs. However, all facilities had the working principle that transfer should be done manually, and the equipment for transfer such as a lifts, a transfer and roller board were not be installed. In changing diapers, bed height adjustment was not possible. And the Japanese standard type of wheelchair has a non-detachable armrest, creating a structural barrier when transferring elderly people from a wheelchair to a toilet seat. At all facilities the basis of care was that caregivers should do it manually. In particular, all facilities had only a weak recognition of the risks of transfer. This investigation shows that facilities for the elderly should rethink elderly care based on a reduction of care load and most importantly protection of caregivers' health. PMID- 16717405 TI - Antibiotic resistance in bacteria and its future for novel antibiotic development. AB - Since the first introduction of the sulfa drugs and penicillin into clinical use, large numbers of antibiotics have been developed and hence contributed to human health. But extensive use of antibiotics has raised a serious public health problem due to multiantibiotic resistant bacterial pathogens that inevitably develop resistance to every new drug launched in the clinic. Consequently, there is a pressing need to develop new antibiotics to keep pace with bacterial resistance. Recent advances in microbial genomics and X-ray crystallography provide opportunities to identify novel antibacterial targets for the development of new classes of antibiotics and to design more potent antimicrobial compounds derived from existing antibiotics respectively. To prevent and control infectious diseases caused by multiantibiotic resistant bacteria, we need to understand more about the molecular aspects of the pathogens' physiology and to pursue ways to prolong the life of precious antibiotics. PMID- 16717404 TI - Enzymatic synthesis of oligosaccharides and neoglycoconjugates. AB - Oligosaccharides involved in glycoconjugates play important roles in a number of biological events. To elucidate the biological functions of oligosaccharides, sufficient quantities of structurally defined oligosaccharides, are of limited availability by traditional purification methods, are required. Hence, chemical and enzymatic syntheses of oligosaccharides are becoming increasingly important in glycobiology and glycotechnology. In addition, oligosaccharides often occur as glycoconjugates attached to proteins or lipids. Hence, the development of simple and effective methods for synthesizing neoglycoconjugates such as neoglycoprotein and neoglycolipids is essential for an understanding of the biological function of these molecules. Here we review the most recent developments in the enzymatic synthesis of oligosaccharides and neoglycoconjugates. PMID- 16717406 TI - Molecular basis for the cellular senescence program and its application to anticancer therapy. AB - Although dysfunctional telomeres and oncogenic or stressful stimuli are known to trigger cellular senescence in normal human diploid cells, the molecules and signaling network involved in the cellular senescence program are not fully understood. We have been trying to identify cellular senescence-inducing factors by various means. First, we screened for an extrinsic signal that can induce cellular senescence in human lung adenocarcinoma cell line A549, and identified transforming growth factor-beta (TGF-beta) as the cellular senescence-inducing factor. Cancer cells senesced by treatment with TGF-beta impaired tumorigenicity both in vitro and in vivo, suggesting that cellular senescence functions as a tumor suppression mechanism. Next, we identified 86 independent senescence associated genes by subtractive screening using A549-derived cell lines. Thirdly, we established novel cell lines (AST cells) from A549 cells exposed to mild oxidative stress. AST cells demonstrated functional impairment of telomerase due to perturbed subcellular localization of human telomerase reverse transcriptase, suggesting that mild oxidative stress might affect the cell fate of cancer cells. These results should provide insight into the molecular basis of the cellular senescence program. PMID- 16717407 TI - Synthetic studies on breviones and structurally related natural products. AB - Breviones, allelopathic agents that have been isolated from Penicillium sp., are structurally unique diterpenoid derivatives. Breviones have attracted attention due to their bioactivity, because their allelopathic activities may offer agricultural use as environmentally benign herbicides. On the other hand, their structural uniqueness is also remarkable, and construction of their unique structure is a challenge from the viewpoint of organic synthesis. This review summarizes synthetic studies on breviones and structurally related natural products. PMID- 16717408 TI - Cloning and characterization of the HPr kinase/phosphorylase gene from Bacillus stearothermophilus No. 236. AB - The Bacillus stearothermophilus no. 236 gene encoding the bifunctional enzyme HprK/P, the key regulator of carbon catabolite repression/activation (CCR/CCA) in most Gram-positive bacteria, was cloned and the (His)(6)-tagged gene product was characterized in detail. The nucleotide sequence of the hprK/P gene corresponded to an open reading frame of 951 bp that encoded a polypeptide of 316 amino acid residues with a calculated molecular mass of 35,458 Da. The deduced amino acid sequence of the B. stearothermophilus no. 236 HprK/P showed 64.5% identity with the B. subtilis enzyme, allowing us to identify two highly conserved motifs, the nucleotide binding P-loop (Walker motif A) and the HprK/P family signature sequence in the C-terminal half of the protein. Furthermore, complementation experiments showed that the cloned hprK/P gene product was functionally active in the B. subtilis cells. The purified (His)(6)-tagged B. stearothermophilus no. 236 HprK/P migrated on SDS-PAGE gel as a single species with a molecular mass of about 36 kDa, and behaved in gel filtration like a hexameric protein. The recombinant protein catalyzes the pyrophosphate (PPi)-dependent (highest activity at pH 7.0 and 40 degrees C) as well as the ATP-dependent phosphorylation of Ser46 in HPr (maximum activity at pH 8.0 and 45 degrees C). It also catalyzes the inorganic phosphate-dependent dephosphorylation (phosphorolysis) of seryl phosphorylated HPr, optimally at pH 6.5 and 40 degrees C. BIAcore surface resonance analysis confirmed that a divalent cation, preferentially Mg(2+), was an indispensable cofactor for the three activities of the HprK/P. Fructose-1,6 bisphosphate (FBP) was observed to stimulate ATP-dependent kinase activity, while inorganic phosophate (Pi) inhibited ATP-dependent kinase activity. Mutations in the Walker motif A simultaneously abolished both types of kinase and phosphorylase activities. On the other hand, the conserved signature residues were confirmed to be involved in the PPi-dependent kinase and phosphorylase reactions. PMID- 16717409 TI - Purification and characterization of fumarase from Corynebacterium glutamicum. AB - Fumarase (EC 4.2.1.2) from Corynebacterium glutamicum (Brevibacterium flavum) ATCC 14067 was purified to homogeneity. Its amino-terminal sequence (residues 1 to 30) corresponded to the sequence (residues 6 to 35) of the deduced product of the fumarase gene of C. glutamicum (GenBank accession no. BAB98403). The molecular mass of the native enzyme was 200 kDa. The protein was a homotetramer, with a 50-kDa subunit molecular mass. The homotetrameric and stable properties indicated that the enzyme belongs to a family of Class II fumarase. Equilibrium constants (K(eq)) for the enzyme reaction were determined at pH 6.0, 7.0, and 8.0, resulting in K(eq)=6.4, 6.1, and 4.6 respectively in phosphate buffer and in 16, 19, and 17 in non-phosphate buffers. Among the amino acids and nucleotides tested, ATP inhibited the enzyme competitively, or in mixed-type, depending on the buffer. Substrate analogs, meso-tartrate, D-tartrate, and pyromellitate, inhibited the enzyme competitively, and D-malate in mixed-type. PMID- 16717410 TI - Purification and characterization of a Cl- -activated aminopeptidase from bovine skeletal muscle. AB - To elucidate the mechanisms involved in the increase in free amino acids during postmortem storage of meat, a novel aminopeptidase was purified from bovine skeletal muscle by ammonium sulfate fractionation and successive chromatographies such as DEAE-cellulose, Sephacryl S-200, Hydroxyapatite, Phenyl-Sepharose, and Hi Trap affinity column chromatography. The molecular mass of the enzyme was found to be 58 kDa on SDS-PAGE. This enzyme had optimum pH at around 7.5, and preferably hydrolyzed Ala-beta-naphthylamide (-NA) in amino acid-NAs. The activity was strongly inhibited by phenylmethansulfonyl fluoride (PMSF) and bestatin, suggesting that it is to be classified as a serine protease. Moreover, the activity was enhanced by chloride and nitrate ions, which is the most remarkable property of this enzyme. The enzyme appeared to be involved in the increase in free amino acids during postmortem storage of meat. PMID- 16717411 TI - Antihypertensive effect of angiotensin I-converting enzyme inhibitory peptides from a sesame protein hydrolysate in spontaneously hypertensive rats. AB - Sesame peptide powder (SPP) exhibited angiotensin I-converting enzyme (ACE) inhibitory activity, and significantly and temporarily decreased the systolic blood pressure (SBP) in spontaneously hypertensive rats (SHRs) by a single administration (1 and 10 mg/kg). Six peptide ACE inhibitors were isolated and identified from SPP. The representative peptides, Leu-Val-Tyr, Leu-Gln-Pro and Leu-Lys-Tyr, could competitively inhibit ACE activity at respective Ki values of 0.92 microM, 0.50 microM, and 0.48 microM. A reconstituted sesame peptide mixture of Leu-Ser-Ala, Leu-Gln-Pro, Leu-Lys-Tyr, Ile-Val-Tyr, Val-Ile-Tyr, Leu-Val-Tyr, and Met-Leu-Pro-Ala-Tyr according to their content ratio in SPP showed a strong antihypertensive effect on SHR at doses of 3.63 and 36.3 microg/kg, which accounted for more than 70% of the corresponding dosage for the SPP-induced hypotensive effect. Repeated oral administration of SPP also lowered both SBP and the aortic ACE activity in SHR. These results demonstrate that SPP would be a beneficial ingredient for preventing and providing therapy against hypertension and its related diseases. PMID- 16717412 TI - Identification of a catalytic residue of Clostridium paraputrificum N-acetyl-beta D-glucosaminidase Nag3A by site-directed mutagenesis. AB - Clostridium paraputrificum M-21 beta-N-acetylglucosaminidase 3A (Nag3A) is an enzyme classified in family 3 of the glycoside hydrolases. To identify catalytic residues of this enzyme, mutations were introduced into highly conserved Glu and Asp residues. Replacement of Asp175 with Ala abolished the catalytic activity without change in the circular dichroism spectrum, strongly suggesting that this residue is a catalytic residue, a nucleophile/base or a proton donor. Since the K(m) values of mutant enzymes D119N, D229N, D229A and D274N increased 17 to 41 times as compared with that of wild-type enzyme, Asp119, Asp229, and Asp274 appear to be involved in substrate recognition and binding. Taking previous studies into consideration, we presume that Asp303 is the catalytic nucleophile and Asp175 is the proton donor of C. paraputrificum Nag3A. PMID- 16717413 TI - Regulation of the body fat percentage in developmental-stage rats by methylxanthine derivatives in a high-fat diet. AB - We investigated the regulatory effects of structural differences among methylxanthine derivatives on the elevation of body fat percentage in developmental-stage rats. Caffeine, theophylline and theobromine were used as the methylxanthines. High-fat diets (20% lard) containing each methylxanthine (0.025%) were administered to male Sprague-Dawley rats for 12 weeks, with the result that the body fat percentage was generally reduced in each methylxanthine fed group. The abdominal adipose tissue weight in the caffeine group was also significantly lower than that in the control group, the serum cholesterol and triglyceride levels in the caffeine group also being significantly lower than the levels in the control group. The study results suggest that caffeine could contribute most to preventing arteriosclerotic diseases. PMID- 16717414 TI - Overexpression and characterization of a carboxypeptidase from the hyperthermophilic archaeon Thermococcus sp. NA1. AB - Genomic analysis of a hyperthermophilic archaeon, Thermococcus sp. NA1, revealed the presence of an 1,497 bp open reading frame, encoding a protein of 499 amino acids. The deduced amino acid sequence was similar to thermostable carboxypeptidase 1 from Pyrococcus furiosus, a member of peptidase family M32. Five motifs, including the HEXXH motif with two histidines coordinated with the active site metal, were conserved. The carboxypeptidase gene was cloned and overexpressed in Escherichia coli. Molecular masses assessed by SDS-PAGE and gel filtration were 61 kDa and 125 kDa respectively, which points to a dimeric structure for the recombinant enzyme, designated TNA1_CP. The enzyme showed optimum activity toward Z-Ala-Arg at pH 6.5 and 70-80 degrees C (k(cat)/K(m)=8.3 mM(-1) s(-1)). In comparison with that of P. furiosus CP (k(cat)/K(m)=667 mM(-1) s(-1)), TNA1_CP exhibited 80-fold lower catalytic efficiency. The enzyme showed broad substrate specificity with a preference for basic, aliphatic, and aromatic C-terminal amino acids. This broad specificity was confirmed by C-terminal ladder sequencing of porcine N-acetyl-renin substrate by TNA1_CP. PMID- 16717415 TI - The effect of pyruvate decarboxylase gene knockout in Saccharomyces cerevisiae on L-lactic acid production. AB - A plant- and crop-based renewable plastic, poly-lactic acid (PLA), is receiving attention as a new material for a sustainable society in place of petroleum-based plastics. We constructed a metabolically engineered Saccharomyces cerevisiae that has both pyruvate decarboxylase genes (PDC1 and PDC5) disrupted in the genetic background to express two copies of the bovine L-lactate dehydrogenase (LDH) gene. With this recombinant, the yield of lactate was 82.3 g/liter, up to 81.5% of the glucose being transformed into lactic acid on neutralizing cultivation, although pdc1 pdc5 double disruption led to ineffective decreases in cell growth and fermentation speed. This strain showed lactate productivity improvement as much as 1.5 times higher than the previous strain. This production yield is the highest value for a lactic acid-producing yeast yet reported. PMID- 16717416 TI - Effects of the principal nutrients on lovastatin production by Monascus pilosus. AB - Lovastatin production is dependent on the substrates provided. We investigated how several carbon and nitrogen sources in the medium affect lovastatin production by Monascus pilosus. M. pilosus required a suitable concentration of organic nitrogen peptone for high lovastatin production. As sole carbon source with peptone, although glucose strongly repressed lovastatin production, maltose was responsible for high production. Interestingly, glycerol combined with maltose enhanced lovastatin production, up to 444 mg/l in the most effective case. Moreover, an isolated mutant, in which glucose repression might be relieved, easily produced the highest level of lovastatin, 725 mg/l on glucose glycerol-peptone medium. These observations indicate that lovastatin production by M. pilosus is regulated by strict glucose repression and that an appropriate release from this repression by optimizing medium composition and/or by a mutation(s) is required for high lovastatin production. PMID- 16717417 TI - Accumulation of gamma-aminobutyric acid in rice germ using protease. AB - The accumulation of gamma-aminobutyric acid (GABA) in the rice germ by using protease was investigated. After the proteolytic hydrolysis of germ protein by trypsin, the amount of GABA reached about 2.26 g per 100 g of germ. This demonstrates that the GABA yield could be significantly increased by protease. Our method is efficient and safe for manufacturing health food enriched with GABA. PMID- 16717418 TI - Postmortem metabolic rate and calpain system activities on beef longissimus tenderness classifications. AB - In this study, we adopted a model of tenderness classification in order to determine the factors affecting the tenderness and tenderization characteristics of beef longissimus, using cluster analysis on the basis of Warner-Bratzler shear force and myofibril fragmentation index, at 1, 7, and 14 d. The rate of tenderization was effectively differentiated by pH, R-values, mu-calpain activity, and calpastatin activity. Differences among tenderness classes were generally detected at 3 and 9 h postmortem for metabolic rate, and at 9 and 24 h for the activities of mu-calpain and calpastatin. Early postmortem metabolic rate and calpain system activities were verified as important factors with regard to longissimus tenderization. PMID- 16717419 TI - Effects of storage temperature on the contents of sugars and free amino acids in tubers from different potato cultivars and acrylamide in chips. AB - To clarify the effects of storage temperature on potato components and acrylamide in chips, tubers from five cultivars were stored at various temperatures (2, 6, 8, 10, and 18 degrees C) for 18 weeks, and the contents of sugars, free amino acids in tubers, and acrylamide in chips after frying were analyzed. At temperatures lower than 8 degrees C, the contents of reducing sugars increased markedly in all cultivars, with similar increases in the acrylamide level and dark brown chip color. Free amino acids showed little change at the storage temperatures tested and varied within certain ranges characteristic of each cultivar. The contents of reducing sugars correlated well with the acrylamide level when the fructose/asparagine molar ratio in the tubers was <2. When the fructose/asparagine ratio was >2 by low-temperature storage, the asparagine content, rather than the reducing sugar content, was found to be the limiting factor for acrylamide formation. PMID- 16717420 TI - Improvement of the glaB promoter expressed in solid-state fermentation (SSF) of Aspergillus oryzae. AB - The glucoamylase-encoding gene (glaB) promoter should be very useful for recombinant protein production in solid-state fermentation (SSF) of Aspergillus oryzae. A 97-bp fragment containing the cis-element of the glaB promoter was inserted into the glaA promoter, which was little expressed in SSF. The chimeric promoter showed about a 24-fold increase in promoter activity in SSF. Eight copies of the 97-bp fragment were tandemly fused with the glaB promoter. The improved promoter showed about a 4.6-fold increase in promoter activity in SSF. The glaB gene was overexpressed under control of the improved glaB promoter in SSF. Recombinant glucoamylase production reached about 1524 mg/kg-broth for 2 d. The improved glaB promoter should be very useful for overproduction of a recombinant protein in SSF of A. oryzae. PMID- 16717421 TI - Dietary indigestible components exert different regional effects on luminal mucin secretion through their bulk-forming property and fermentability. AB - The purpose of this study was to examine the effects of dietary indigestible components on mucin secretion in the respective parts of the gastrointestinal tract through their physico-chemical properties. Rats were fed either a control diet or diets containing 5% polystyrene foam (PSF), 5% fructooligosaccharide (FOS), 5% PSF + 5% FOS, or 10% beet fiber for 10 d. Mucins in the small intestine and feces were greater in the PSF, PSF + FOS, and beet fiber groups than in the control and FOS groups. In the cecum, greater mucins were observed in the FOS, PSF + FOS, and beet fiber groups than in the control and PSF groups. None of the dietary treatment was effective on gastric mucins. Cecal mucins were significantly correlated with the cecal pool sizes of total short-chain fatty acids. The correlation between fecal mucins and fecal numbers was also significant. The results suggest that the effect of the bulk-forming property of the dietary indigestible component on mucin secretion is limited to the duct, while fermentability is effective only in the cecum. PMID- 16717423 TI - Rice bifunctional alpha-amylase/subtilisin inhibitor: cloning and characterization of the recombinant inhibitor expressed in Escherichia coli. AB - The complete nucleotide sequences of the cDNA and its gene that encode a bifunctional alpha-amylase/subtilisin inhibitor of rice (Oryza sativa L.) (RASI) were analyzed. RASI cDNA (939 bp) encoded a 200-residue polypeptide with a molecular mass of 21,417 Da, including a signal peptide of 22 amino acids. Sequence comparison and phylogenetic analysis showed that RASI is closely related to alpha-amylase/subtilisin inhibitors from barley and wheat. RASI was found to be expressed only in seeds, suggesting that it has a seed-specific function. A coding region of RASI cDNA without the signal peptide was introduced into Escherichia coli and was expressed as a His-tagged protein. Recombinant RASI was purified to homogeneity in a single step by Ni-chelating affinity column chromatography and characterized to elucidate the target enzyme. The recombinant inhibitor had strong inhibitory activity toward subtilisin, with an equimolar relationship, comparable with that of native RASI, and weak inhibitory activity toward some microbial alpha-amylases, but not toward animal or insect alpha amylases. These results suggest that RASI might function in the defense of the seed against microorganisms. PMID- 16717422 TI - Cloning, expression, and identification of immunological activity of an allergenic protein in tartary buckwheat. AB - A cDNA fragment encoding a 24 kDa allergenic protein in tartary buckwheat was obtained using reverse transcription PCR, 3'-rapid amplification of cDNA ends (RACE), and nest PCR. The cDNA clone contained 768 nucleotides, including 588 nucleotides in the open reading frame (ORF) and 180 nucleotides in the 3' terminal sequence. The ORF encoded a functional protein of 195 amino acids. It shared 95% and 93% nucleotide homology with the allergenic storage protein and a legumin-like protein from common buckwheat respectively. The encoding region was expressed in host strain Escherichia coli BL21 (DE3) induced by IPTG at 28 degrees C. The inclusion bodies of recombinant protein obtained were analyzed by western blot and purified by affinity chromatography. The purity of target protein reached above 95%. After they were refolded by step-wise dialysis, 68% of the inclusion bodies reached soluble state. An analysis of immunological activity showed that the recombinant protein had a specific IgE binding activity. This is the first report of the molecular cloning and expression of the major allergen from tartary buckwheat. PMID- 16717424 TI - Characterization of glycosynthase mutants derived from glycoside hydrolase family 10 xylanases. AB - Four xylanases belonging to glycoside hydrolase family 10-Thermotoga maritima XylB (TM), Clostridium stercorarium XynB (CS), Bacillus halodurans XynA (BH), and Cellulomonas fimi Cex (CF)-were converted to glycosynthases by substituting the nucleophilic glutamic acid residues with glycine, alanine, and serine. The glycine mutants exhibited the highest levels of glycosynthase activity with all four enzymes. All the glycine mutants formed polymeric beta-1,4-linked xylopyranose as a precipitate during reaction with alpha-xylobiosyl fluoride. Two glycine mutants (TM and CF) recognized X(2) as an effective acceptor molecule to prohibit the formation of the polymer, while the other two (CS and BH) did not. The difference in acceptor specificity is considered to reflect the difference in substrate affinity at their +2 subsites. The results agreed with the structural predictions of the subsite, where TM and CF exhibit high affinity at subsite 2, suggesting that the glycosynthase technique is useful for investigating the affinity of +subsites. PMID- 16717425 TI - Partial characterization and immunostimulatory effect of a novel polysaccharide protein complex extracted from Phellinus linteus. AB - Many polysaccharides isolated from mushroom are considered to be biological response modifiers and have been shown to enhance various immune responses in vivo and in vitro. We demonstrate that a novel polysaccharide-protein complex (PPC) extracted from Phellinus linteus was a potent immunomodulator. PPC had a molecular weight of approximately 73 kDa. It was composed of five different monosaccharides, predominantly D-glucose and D-mannose, in the molar ratio of 3:2, the main amino acid being aspartic acid. PPC had a unique mode of immunostimulation with regard to its cell-type specificity. PPC was found to markedly increase the proliferation of B cells, but not T cells. Although PPC and lipopolysaccharide (LPS) had a similar mode of action in B cells, they were differentiated by the fact that PPC-induced cellular activation was not inhibited by polymyxin B (PB), a specific inhibitor of LPS. PPC increased the cytokine production and nitric oxide (NO) from macrophages. PPC also enhanced the lytic death of NO-sensitive tumor cells, B16 melanoma, through the production of NO. In addition, PPC up-regulated the natural killer (NK) cell-mediated killing of tumor cells, YAC-1 lymphoma in vitro. These results suggest that PPC stimulated the tumoricidal activities of macrophages and NK cells, and induced the proliferation of B cells in vitro. This process may be the mechanism by which PPC produced its therapeutic effects. PMID- 16717426 TI - Reduction of allergenic proteins by the effect of the ripening inhibitor (rin) mutant gene in an F1 hybrid of the rin mutant tomato. AB - The ripening inhibitor (rin) mutant tomato yields non-ripening fruit, and the rin hybrid fruit (RIN/rin) shows an intermediate phenotype between the wild and mutant fruit, that is, red-ripe and extended shelf life. We found by a microarray analysis that the genes encoding possible allergenic proteins were expressed at a significantly lower level in the rin hybrid fruit than in the wild-type fruit. These allergenic proteins, which were beta-fructofuranosidase and polygalacturonase 2A (PG-2A), were confirmed to accumulate at a lower level in the rin hybrid fruit than in the wild-type fruit. The immunoglobulin E (IgE) in serum from a tomato-allergic patient showed lower reactivity to the extract of the rin hybrid fruit than to that of the wild fruit. These results suggest that the rin gene has the potential to regulate allergen accumulation in tomato fruit. PMID- 16717427 TI - Yeast cell death caused by mutation of the OST2 gene encoding the epsilon-subunit of Saccharomyces cerevisiae oligosaccharyltransferase. AB - An essential epsilon-subunit of oligosaccharyltransferase Ost2 is a yeast homolog of mammalian highly conserved DAD1 (defender against apoptotic death). In hamster cells, the Gly38Arg mutation in DAD1 causes apoptosis at restrictive temperatures due to a defect in N-linked glycosylation. To analyze the function of Ost2 in yeast cell death, we constructed Saccharomyces cerevisiae strains expressing Gly58Arg (corresponding to the Gly38Arg mutation in hamster DAD1), Gly86Arg, and Glu113Val mutant Ost2. At elevated temperatures, ost2 mutants arrested growth by decreasing cell viability. Phosphatidylserine exposure, a phenotypic marker of apoptosis in mammalian cells, was found in ost2 mutant cells at 37 degrees C, although DNA fragmentation was not clearly detected. A high concentration of sorbitol compensates for the temperature sensitivity of the ost2 mutant. These results suggest that apoptosis-like cell death in ost2 mutants is caused by the secondary effect of overall reduced protein N-linked glycosylation. PMID- 16717429 TI - Degradation of PrP(Sc) by keratinolytic protease from Nocardiopsis sp. TOA-1. AB - A keratinolytic alkaline proteae (NAPase) from Nocardiopsis sp. TOA-1 degraded a scrapie prion without any chemical or physical treatment. Optimal temperature and pH were 60 degrees C and above pH 10.0. The scrapie prion was completely degraded within 3 min under optimal conditions. PMID- 16717428 TI - Down-regulation of mdr1b mRNA expression in the kidneys of mice following maternal exposure to tributyltin chloride. AB - We investigated the change in renal mdr1b mRNA expression in offspring exposed to tributyltin chloride (TBTC) via the placenta and lactation or via lactation, using the real-time reverse transcription-polymerase chain reaction. Pregnant ICR mice were given water containing TBTC (0, 15, and 50 microg/ml) ad libitum from the start of pregnancy to weaning or from parturition to weaning. Exposure via the placenta and lactation significantly reduced the renal mdr1b level in offspring. Exposure to TBTC through the mother might impair the exclusion system of toxic compounds in offspring. PMID- 16717430 TI - Changes in the immunogold electron-microscopic localization of calpain in bovine skeletal muscle induced by conditioning and high-pressure treatment. AB - Changes in the localization of calpain in conditioned and pressurized bovine skeletal muscles were investigated by immunogold electron-microscopy. In the muscle immediately after thawing (control), the relative distributions of colloidal particles statistically calculated by counting the colloidal particles were about 65% and 35% in the I-band/Z-disk and A-band regions, respectively. In the muscle conditioned for 7 days, distribution of colloidal particles was more than two times greater in both the I-band/Z-disk and A-band regions than in the control muscle. Almost no change in either the absolute concentration or relative distribution of the colloidal particles was detectable during further storage. In the muscle exposed to a pressure of 100 MPa or 200 MPa, slightly more immunogold was detected in the sarcomere than in that of the control muscle. Increasing pressure up to 300 MPa enabled high-density particles to be seen throughout the sarcomere. Conversely, few particles were detected in the sarcomere of the muscle exposed to 400 MPa. These electron-microscopic observations were confirmed from the statistical analysis as with the conditioned muscles. It was clear from the results obtained that the pressure-induced changes in calpain localization were much more pronounced than those from conditioning. PMID- 16717431 TI - Effect of Dahi containing Lactococcus lactis on the progression of diabetes induced by a high-fructose diet in rats. AB - The antidiabetic effect of dahi was observed on high-fructose-induced diabetic rats. The fasting blood glucose, glycosylated haemoglobin, insulin, free fatty acids and triglyceride levels of the dahi fed group animals were significantly lower than those of the control group (p<0.05). The imparity of the glucose tolerance test was also delayed by one week in the dahi-fed animals. PMID- 16717432 TI - Biotransformation of (-)-menthone by human liver microsomes. AB - The aim of the current study was to investigate the metabolism of (-)-menthone by liver microsomes of humans. (-)-Menthone (1) was metabolized to (+)-neomenthol (2) (3-reduction) and 7-hydroxymenthone (3) by human liver microsomes. The metabolites formed were analyzed on GC and GC-MS. Kinetic analysis showed that K(m) and V(max) values for the metabolized (-)-menthone to respective (+) neomenthol and 7-hydroxymenthone by liver microsomes of human sample HG70 were 0.37 mM and 4.91 nmol/min/mg protein and 0.07 mM and 0.71 nmol/min/mg protein. PMID- 16717433 TI - Interaction of subtilisin BPN' and recombinant fungal protease inhibitor F from silkworm with substituted P1 site residues. AB - Fungal protease inhibitor F (FPI-F) from silkworm inhibits subtilisin and fungal proteases. FPI-F mutants P(1) residues of which, Thr(29), were replaced with Glu, Phe, Gly, Leu, Met, and Arg, were prepared. The inhibitory activities of mutated FPI-F against subtilisin and other mammalian proteases indicated that FPI-F might be a specific inhibitor toward subtilisin-type protease. PMID- 16717434 TI - Identification of 2-(cysteinyl)amido-2-deoxy-D-galacturonic acid residue from the sheath of Leptothrix cholodnii. AB - The sheath of Leptothrix cholodnii is a glycoconjugate composed of a polysaccharide and a peptide rich in cysteine. In this study, structural determination of the hydrazinolyzate of the sheath was carried out. Since the hydrazinolyzate is a polysaccharide incorporated with cysteine, it was S derivatized with a thiol-specific fluorogenic reagent, 4-(aminosulfonyl)-7-fluoro 2,1,3-benzoxadiazole (ABD-F). Fluorescent fragments were purified by HPLC, and their structures were analyzed by mass spectrometry and NMR spectroscopy. The sheath was found to contain 2-(cysteinyl)amido-2-deoxy-D-galacturonic acid residue. PMID- 16717436 TI - Decomposition reaction of sesamin in supercritical water. AB - The methylenedioxyphenyl moiety in the structure of sesamin and episesamin was changed into the catechol moieties, (1R,2S,5R,6S)-6-(3,4-dihydroxyphenyl)-2-(3,4 methylenedioxyphenyl)-3,7-dioxabicyclo[3,3,0]octane, (1R,2R,5R,6S)-2-(3,4 dihydroxyphenyl)-6-(3,4-methylenedioxyphenyl)-3,7-dioxabicyclo[3,3,0]octane, (1R,2R,5R,6S)-6-(3,4-dihydroxyphenyl)-2-(3,4-methylenedioxyphenyl)-3,7 dioxabicyclo[3,3,0]octane, (1R,2S,5R,6S)-2,6-bis(3,4-dihydroxyphenyl)-3,7 dioxabicyclo[3,3,0]octane, and (1R,2R,5R,6S)-2,6-bis(3,4-dihydroxyphenyl)-3,7 dioxabicyclo[3,3,0]octane, in supercritical water. These products had same structures as the sesamin metabolites which act as antioxidants in the liver. These features suggested the direct preparation of antioxidants from sesamin by a one-step reaction using supercritical water. PMID- 16717435 TI - Production of three O-methhylated esculetins with Escherichia coli expressing O methyltransferase from poplar. AB - O-Methyltransferase, POMT-9 was expressed in Escherichia coli. HPLC analysis of reaction products revealed three peaks corresponding to isoscopoletin, scopoletin, and scoparone, and their structures were determined using NMR. Biotransformation of esculetin with E. coli expressing POMT-9 generated scopoletin, isoscopoletin, and scoparone at 30.3, 21, and 31 microM respectively. POMT-9 is the first O-methyltransferase that produces three different O methylated products. PMID- 16717437 TI - Restriction of the in vitro formation of angiotensin II by leucinyl-arginyl tryptophan, a novel peptide with potent angiotensin I-converting enzyme inhibitory activity. AB - Leucinyl-arginyl-tryptophan (LRW) is a new peptide inhibitor of the angiotensin converting enzyme (ACE) that was previously predicted through quantitative structure-activity relationship modeling. LRW inhibited ACE activity in a competitive manner with a higher K(m) value in the presence of the peptide, and the in vitro formation of angiotensin II by ACE was significantly reduced in the presence of LRW up to 60 min of incubation time. PMID- 16717438 TI - Hepatoprotective effects of whey protein on D-galactosamine-induced hepatitis and liver fibrosis in rats. AB - The hepatoprotective effects of whey protein on two injections of D-galactosamine (300 mg/kg, i.p.) were investigated in rats fed a modified AIN-93M diet formulated with a protein source of casein or whey for 16 d. The whey protein containing diet clearly suppressed an increase in plasma alanine and aspartate aminotransferase activity, lactate dehydrogenase and bilirubin, which are hepatitis markers, and also hyaluronic acid, a fibrosis marker. In addition, it suppressed histopathological signs of portal fibrosis, bile duct proliferation, and perivenular sclerosis. These results suggest that supplementation with whey protein can help prevent the development of hepatitis and portal fibrosis. PMID- 16717439 TI - Aortic stiffness, flow-mediated dilatation and carotid intima-media thickness in obstructive sleep apnea: non-invasive indicators of atherosclerosis. AB - BACKGROUND AND OBJECTIVE: Obstructive sleep apnea (OSA) has a critical association with cardiovascular mortality and morbidity. Carotid intima-media thickness (IMT), flow-mediated dilatation (FMD) and aortic stiffness are early signs of atherosclerosis. The presence of subclinical atherosclerosis was assessed in OSA patients using these parameters. METHODS: 40 patients with OSA showing an apnea-hypopnea index (AHI) > or =5 (mean age 51.3 +/- 9 years, 32 males) and 24 controls (AHI < 5, mean age 51.9 +/- 5.2 years, 19 males) were enrolled in the study. In all subjects, polysomnographic examination and recordings were performed during sleep. IMT of the carotid artery, endothelium dependent/-independent vasodilation of the brachial artery and aortic elastic parameters were investigated using high-resolution Doppler echocardiography. RESULTS: The demographic data of the patients with OSA and controls were not significantly different. Subjects with OSA demonstrated higher values of aortic stiffness (7.1 +/- 1.88 vs. 6.42 +/- 1.56, respectively) and IMT (0.85 +/- 0.13 vs. 0.63 +/- 0.11 mm, p = 0.0001, respectively) but lower distensibility (9.47 +/ 1.33 vs. 11.8 +/- 3.36 cm(2)/dyn/10(6)) and FMD (4.57 +/- 1.3 vs. 6.34 +/- 0.83%, p = 0.0001, respectively) than the controls. The respiratory disturbance index correlated positively with aortic stiffness and IMT and negatively with distensibility and FMD. CONCLUSION: We observed blunted endothelium-dependent dilatation, increased carotid IMT and aortic stiffness in patients with OSA compared with matched control subjects. This is evident in the absence of other diseases, suggesting that OSA is an independent cause of atherosclerosis. These simple and non-invasive methods help to detect subclinical atherosclerosis in OSA. PMID- 16717440 TI - Phenotypic characterization of DFNA24: prelingual progressive sensorineural hearing impairment. AB - This article describes the hearing impairment (HI) phenotype which segregates in a large multi-generation Swiss-German family with autosomal dominant nonsyndromic HI. The locus segregating within this pedigree is located on chromosome 4q35-qter and is designated as DFNA24. For this pedigree, audiometric data on 25 hearing impaired family members are available. It was demonstrated that within this kindred the HI is sensorineural, bilateral, prelingual in onset, and progressive throughout life. Age-related typical audiograms depict steeply down-sloping curves, with moderate high-frequency HI at birth, then steady progression to moderate HI in the low frequencies, severe HI at mid-frequencies and profound HI at high frequencies by age 70. Annual threshold deterioration was approximately 0.5 dB/year at 1-2 kHz after correction for presbycusis. PMID- 16717441 TI - A modified version of tinnitus retraining therapy: observing long-term outcome and predictors. AB - Tinnitus retraining therapy (TRT) in Germany includes not only directive counseling and sound therapy but also stress management and facultative psychotherapeutic treatment. The aim of the present study was to investigate the impact of this modified version of TRT on certain tinnitus-related aspects of distress and variables that may predict treatment outcome. Clinical data from 92 patients undergoing outpatient TRT in the Charite Tinnitus Centre were evaluated retrospectively over 1 year. Treatment outcome was defined by changes in specific areas of tinnitus-related distress and assessed by the Tinnitus Questionnaire. Changes in audiometric frequency and loudness of tinnitus were examined by regular audiometric testing. The overall Tinnitus Questionnaire score was significantly reduced after 1 year. Severely affected tinnitus sufferers (decompensated tinnitus) profited more than less affected patients (compensated tinnitus). In cases of indicated psychotherapy, improvement was significant for the patients who took advantage of psychotherapeutic treatment during TRT but was not significant for those who interrupted or dismissed an indicated psychotherapy. Changes in tinnitus-specific areas of distress were most pronounced in the scales for emotional and cognitive distress and intrusiveness. Significant changes in sleep disturbances, auditory perceptual difficulties and somatic complaints were observed in patients with decompensated tinnitus. In patients with chronic tinnitus, modified TRT may lead to significant subjective improvement in certain tinnitus-related symptoms like emotional and cognitive distress and intrusiveness. Particularly patients suffering from severe tinnitus distress take advantage of therapy. Careful psychotherapeutic diagnostics and therapies and, if necessary, motivation to make use of psychotherapy seem to be essential preconditions for therapeutic success in patients with severe psychosomatic comorbidity. PMID- 16717442 TI - Comparative morphology of the avian cerebellum: I. Degree of foliation. AB - Despite the conservative circuitry of the cerebellum, there is considerable variation in the shape of the cerebellum among vertebrates. One aspect of cerebellar morphology that is of particular interest is the degree of folding, or foliation, of the cerebellum and its functional significance. Here, we present the first comprehensive analysis of variation in cerebellar foliation in birds with the aim of determining the effects that allometry, phylogeny and development have on species differences in the degree of cerebellar foliation. Using both conventional and phylogenetically based statistics, we assess the effects of these variables on cerebellar foliation among 91 species of birds. Overall, our results indicate that allometry exerts the strongest effect and accounts for more than half of the interspecific variation in cerebellar foliation. In addition, we detected a significant phylogenetic effect. A comparison among orders revealed that several groups, corvids, parrots and seabirds, have significantly more foliated cerebella than other groups, after accounting for allometric effects. Lastly, developmental mode was weakly correlated with relative cerebellar foliation, but incubation period and fledging age were not. From our analyses, we conclude that allometric and phylogenetic effects exert the strongest effects and developmental mode a weak effect on avian cerebellar foliation. The phylogenetic distribution of highly foliated cerebella also suggests that cognitive and/or behavioral differences play a role in the evolution of the cerebellum. PMID- 16717443 TI - Effects of midazolam and morphine on cerebral oxygenation and hemodynamics in ventilated premature infants. AB - BACKGROUND: Midazolam sedation and morphine analgesia are commonly used in ventilated premature infants. OBJECTIVES: To evaluate the effects of midazolam versus morphine infusion on cerebral oxygenation and hemodynamics in ventilated premature infants. METHODS: 11 patients (GA 26.6-33.0 weeks, BW 780-2,335 g) were sedated with midazolam (loading dose 0.2 mg/kg, maintenance 0.2 mg/kg/h) and 10 patients (GA 26.4-33.3 weeks, BW 842-1,955 g) were sedated with morphine (loading dose 0.05 mg/kg, maintenance 0.01 mg/kg/h). Changes in oxyhemoglobin (Delta cO2Hb) and deoxyhemoglobin (Delta cHHb) were assessed using near infrared spectrophotometry. Changes in cHbD (= Delta cO(2)Hb - Delta cHHb) reflect changes in cerebral blood oxygenation and changes in concentration of total hemoglobin (Delta ctHb = Delta cO2Hb + Delta cHHb) represent changes in cerebral blood volume (DeltaCBV). Changes in cerebral blood flow velocity (DeltaCBFV) were intermittently measured using Doppler ultrasound. Heart rate (HR), mean arterial blood pressure (MABP), arterial oxygen saturation (saO2) and transcutaneous measured pO2 (tcpO2) and pCO2 (tcpCO2) were continuously registered. Statistical analyses were carried out using linear mixed models to account for the longitudinal character study design. RESULTS: Within 15 min after the loading dose of midazolam, a decrease in saO2, tcpO2 and cHbD was observed in 5/11 infants. In addition, a fall in MABP and CBFV was observed 15 min after midazolam administration. Immediately after morphine infusion a decrease in saO2, tcpO2 and cHbD was observed in 6/10 infants. Furthermore, morphine infusion resulted in a persistent increase in CBV. CONCLUSIONS: Administration of midazolam and morphine in ventilated premature infants causes significant changes in cerebral oxygenation and hemodynamics, which might be harmful. PMID- 16717445 TI - Classification of DNA methylation patterns in tumor cell genomes using a CpG island microarray. AB - Our group has initiated experiments to epigenetically profile CpG island hypermethylation in genomic DNA from tissue specimens of head and neck squamous cell carcinoma (HNSCC) using a microarray of 12,288 CpG island clones. Our technique, known as a methylation-specific restriction enzyme (MSRE) analysis, is a variation of the differential methylation hybridization (DMH) technique, in that it is not an array comparison of two DNA samples using methylation-specific restriction enzymes. Instead, it is a comparison of a single DNA sample's response to a methylation-sensitive restriction enzyme (HpaII) and its corresponding methylation-insensitive isoschizomer (MspI). Estimation of the reproducibility of this microarray assay by intraclass correlation (ICC) demonstrated that in four replicate experiments for three tumor specimens, the ICC observed for a given tumor specimen ranged from 0.68 to 0.85 without filtering of data. Repeated assays achieved 87% concordance or greater for all tumors after filtering of array data by fluorescence intensity. We utilized hierarchical clustering on a population of 37 HNSCC samples to cluster tumor samples with similar DNA methylation profiles. Supervised learning techniques are now being utilized to allow us to identify associations between specific epigenetic signatures and clinical parameters. Such techniques will allow us to identify select groups of CpG island loci that could be used as epigenetic markers for both diagnosis and prognosis in HNSCC. PMID- 16717446 TI - SLUG (SNAI2) overexpression in embryonic development. AB - The Snail-related zinc-finger transcription factor, SLUG (SNAI2), is critical for the normal development of neural crest-derived cells and loss-of-function SLUG mutations have been proven to cause piebaldism and Waardenburg syndrome type 2 in a dose-dependent fashion. However, little is known about the consequences of SLUG overexpression in embryonic development. We report SLUG duplication in a child with a unique de novo 8q11.2-->q13.3 duplication associated with tetralogy of Fallot, submucous cleft palate, renal anomalies, hypotonia and developmental delay. To investigate the effects of Slug overexpression on development, we analyzed mice carrying a Slug transgene. These mice were morphologically normal at birth, inferring that Slug overexpression is not sufficient to cause overt morphogenetic defects. In the adult mice, there was a 20% incidence of sudden death, cardiomegaly and cardiac failure associated with incipient mesenchymal tumorigenesis. These findings, while not directly implicating Slug in congenital and acquired heart disease, raise the possibility that Slug overexpression may contribute to specific cardiac phenotypes and cancer development. PMID- 16717447 TI - Increased susceptibility to maternal aneuploidy demonstrated by comparative genomic hybridization analysis of human MII oocytes and first polar bodies. AB - Single cell comparative genomic hybridization (CGH) was employed to extensively investigate 24 unfertilized or in vitromatured meiosis II oocytes and their corresponding first polar bodies (PBs), to determine how and whether all 23 chromosomes participate in female meiosis I errors and to accurately estimate the aneuploidy rate in the examined cells. Results were obtained for 15 oocytes and 16 PBs, representing 23 eggs (MII oocyte-PB complexes) donated from 15 patients (average age 32.2 years). Abnormalities were detected in ten eggs, giving an overall aneuploidy rate of 43.5%. In all, fourteen anomalies were scored, with the fertilized oocyte being at risk of monosomy in eight cases and at risk of trisomy in six; chromosomes of various sizes participated. CGH was able to give a comprehensive aneuploidy rate, as both absence of chromosomal material and the presence of extra copies were accurately scored. The aneuploidy mechanisms determined were: classical whole univalent non-disjunction; chromatid predivision prior to anaphase I, leading to metaphase II imbalance. There was also evidence of germinal mosaicism for a trisomic cell line. Three patients appeared to be predisposed to meiosis I errors, based on the presence of either multiple abnormalities in one or more of their examined cells, or of the same type of abnormality in all of their cells. Exclusion of these susceptible patients reduces the aneuploidy rate to 20%. Various hypotheses are put forward to explain these observations in order to stimulate research into the complex nature of female meiotic regulation. PMID- 16717444 TI - Applying whole-genome studies of epigenetic regulation to study human disease. PMID- 16717448 TI - Isolation, mapping and identification of SNPs for four genes (ACP6, CGN, ANXA9, SLC27A3) from a bovine QTL region on BTA3. AB - On the basis of fine mapping of a quantitative trait loci region of BTA3 for milk fat content, an examination of the comparative map between cattle and human indicates that the annexin 9 protein gene (ANXA9) and the fatty acid transport protein type 3 gene (SLC27A3) are two strong candidate genes. The objective of the present study is to isolate, map and characterize these genes and identify polymorphisms that could be further utilized in linkage or association studies. Furthermore, two new genes which are in the same region, cingulin protein gene (CGN) and lysophosphatidic acid phosphatase protein gene (ACP6) were studied. DNA fragments (869, 1778, 1933 and 2618 bp) corresponding to partial sequences of ACP6,CGN,ANXA9 and SLC27A3 genes were isolated. Direct sequencing of PCR products amplified from different cattle breeds revealed 1, 4, 4 and 2 SNPs for ACP6, CGN,ANXA9 and SLC27A3, respectively. For ANXA9 one SNP was located in exon 5 (A- >G 951) resulting in an amino acid change from histidine to arginine. Finally, ACP6,CGN,ANXA9 and SLC27A3 genes were located on chromosome 3 between ILSTS096 and BMS819 markers, in a region in which quantitative trait loci (QTL) for several milk traits have been described. PMID- 16717449 TI - Cloning, chromosome mapping and expression characteristics of porcine ANGPTL3 and -4. AB - Angiopoietin-like protein 3 and -4 (ANGPTL3 and -4) are two members of angiopoietin-like proteins (ANGPTLs), which have the signature structure of the angiopoietin family but cannot bind to the TIE2 receptor. It has been reported that they both affect lipid metabolism by inhibiting the activity of lipoprotein lipase (LPL). Here we report the cDNA cloning, chromosome mapping and expression analysis of ANGPTL3 and -4 in pigs. Sequence analysis shows that ANGPTL3 contains an open reading frame of 1,389 bp, which encodes 462 amino acids, and ANGPTL4 contains a coding region of 1,239 bp, which encodes 412 amino acids. Porcine ANGPTL3 deduced amino acid sequence shares 83% and 73.7% identity with human and mouse, respectively, and ANGPTL4 shares 79.4% and 77.7% amino acid identity with human and mouse, respectively. Porcine ANGPTL3 and -4 were mapped to the 6q31- >q35 and 2q21-->q24 region, respectively, by radiation hybrid mapping. Tissue distribution analysis indicated that porcine ANGPTL3 mRNA was exclusively expressed in liver, and porcine ANGPTL4 was ubiquitously expressed with the highest abundance in white adipose tissue. Furthermore, the mRNA level of ANGPTL3 and -4 in liver and the mRNA level of ANGPTL4 in white adipose tissue were significantly higher in genetically obese pigs than in their lean counterparts. This is the first report of molecular cloning and characterization of ANGPTL3 and -4 in pigs, which will be helpful for a better understanding of the role of ANGPTLs in lipid metabolism. PMID- 16717450 TI - Chromosomal mechanisms underlying the karyotype evolution of the oriental voles (Muridae, Eothenomys). AB - We have investigated the karyotype relationships of two oriental voles, i.e. the Yulong vole (Eothenomys proditor, 2n = 32) and the large oriental vole (Eothenomys miletus, 2n = 56) as well as the Clarke's vole (Microtus clarkei, 2n = 52), by a combined approach of cross-species chromosome painting and high resolution G-banding comparison. Chromosome-specific painting probes were generated from flow-sorted chromosomes of E. proditor and hybridized onto metaphases of E. proditor, E. miletus and M. clarkei, leading to the establishment of genome-wide comparative chromosome maps. Our results demonstrate that Robertsonian translocations (centric fusions) have played a major role in the karyotype evolution of oriental voles with no obvious evidence for the involvement of tandem fusions as proposed previously and that the genome organizations of vole species are highly conserved. The comparative chromosome maps of these three vole species belonging to two phylogenetically distinct genera provide a framework for future studies on the karyotype evolution in voles. PMID- 16717451 TI - Karyotypes of Rana tagoi Okada with diploid number 28 in the Chausu Mountains of the Minamishinshu district of Nagano Prefecture, Japan (Anura: Ranidae). AB - Karyotypes of Tago's brown frog Rana tagoi from the Chausu mountains in Minamishinshu of Nagano Prefecture were examined by conventional Giemsa staining, C-banding and late replication (LR)-banding. Chromosome number was 2n = 28 in all cases. The 28 chromosomes consisted of four pairs (1-4) of large biarmed chromosomes, two pairs (5-6) of telocentric chromosomes and eight pairs (7-14) of small biarmed chromosomes. Chromosome pair 11 had a secondary constriction on the long arm. In females, the C-band on the long arm of chromosome pair 6 was detected in both homologs, but was absent from the arms of the homologs of chromosome pairs 5 and 9. In males, C-bands were found in the long arms of both homologs of chromosome pairs 5 and 6, were present only in one homolog of chromosome pair 5 for certain male specimens and found in only one homolog of chromosome pair 9. Specimens of R. tagoi (2n = 28) should thus have two pairs of telocentric chromosomes to provide the same number of chromosome arms, these originating quite likely from chromosome pair 1 in the 26-chromosome specimens by centric fission. Heteromorphic sex chromosomes of the XX-XY type in R. tagoi (2n = 28) in the Chausu mountains were identified. Karyotypes of tail-tip cells from a hybrid tadpole between female R. tagoi (2n = 26) from the Hinohara village in Tokyo and male R. tagoi (2n = 28) from the Chausu mountain population were examined by squash preparation. Chromosome number was 2n = 27 in all tadpoles. The 27 chromosomes consisted of one chromosome set of R. tagoi (2n = 28) and one of R. tagoi (2n = 26). PMID- 16717452 TI - Detection of B chromosomes in interphase hemolymph nuclei from living specimens of the grasshopper Eyprepocnemis plorans. AB - The two most important evolutionary properties of B chromosomes are their transmission rate (which suggests their selfishness when significantly higher than 0.5) and their net effects on carrier fitness (usually negative for parasitic Bs). The study of transmission rate unavoidably requires the analysis of many controlled crosses in order to accurately measure population average transmission rate. Therefore, getting a marker closely associated to B presence is of crucial importance to alleviate the load of performing many useless crosses between lacking B individuals. After investigating several cytogenetic techniques on several tissues that may be sampled without drastically damaging live specimens of the grasshopper Eyprepocnemis plorans, we report here the excellent results provided by the CMA3 fluorescence and C-banding techniques applied to hemocyte nuclei. These cells may be easily obtained from both males and females and provide information on B presence even during the interphase stage. The two cytogenetic techniques take advantage of the heterochromatic nature of the B chromosomes, so that Bs made predominantly of ribosomal DNA are revealed by CMA3 as bright bodies in the interphase hemocytes, and Bs mostly made of satellite DNA are visualized by C-banding as intensely stained bodies in these cells. PMID- 16717453 TI - Cytological characterization of sex chromosomes and ribosomal DNA location in Anastrepha species (Diptera, Tephritidae). AB - This paper reports a comparative analysis of heterochromatin organization in the sex chromosomes of the fruit fly Anastrepha. Mitotic chromosomes of eight Anastrepha species from different taxonomic groups were stained with DAPI and chromomycin A3 fluorochromes followed by C-banding. A specific sex-chromosome banding pattern was obtained for each of the analyzed species. Fluorescence in situ hybridization (FISH) was performed to investigate the chromosomal location of rDNA loci. In all cases the rDNA sequences were found to localize exclusively to the sex chromosomes. The results further extend the chromosomal knowledge of Anastrepha and allow a precise species identification. PMID- 16717454 TI - Nuclear DNA content and nuclear and cell volume are positively correlated in angiosperms. AB - Volumes of flow sorted nuclei were analyzed from two highly endopolyploid (diploids with endopolyploid tissues) species (Arabidopsis thaliana and Barbarea stricta), from a less endopolyploid species (Allium cepa) and from two non endopolyploid species (Chrysanthemum multicolor and Fritillaria uva-vulpis). Intraspecific as well as interspecific comparisons revealed a highly positive correlation (r > 0.99) between DNA content and nuclear volume. No significant differences between expected and measured nuclear volumes were noted indicating that chromatin packing is not increased with increasing DNA content in the tested plant species. In epidermis cells of A. thaliana, A. cepa and Ch. multicolor, a lower (r between 0.6 and 0.7) but significant positive correlation between nuclear volume and cell volume was found. This correlation is compatible with the hypothesis that endopolyploidization (EP = consecutive replication cycles not separated by nuclear and cell divisions) might speed up the growth of endopolyploid species and compensate for small genome size. PMID- 16717455 TI - A highly complex rea(2;3;11) and aniridia by position effect. AB - A two-year-old boy presenting with bilateral aniridia and psychomotor retardation had a de novo (2;3;11) highly complex rearrangement which was characterized as far as possible by means of G-banding and FISH assays with multiple probes including cosmids for the Wilms, Aniridia, Genital anomalies and Retardation (WAGR) region, alphoid repeats for chromosomes 2, 3 and 11, subtelomere probes for 2p/2q, 3p/3q and 11q and BACs for 2q32 and 3q13. We identified approximately 15 breakpoints with at least three interchromosomal and three intrachromosome anomalies involving chromosome 11. Both parents had normal karyotypes and no cryptic 11p rearrangements revealed by the chromosome 11 cosmid panel. The lack of a deletion of PAX6 pointed to the direct insertion of an approximately 300-kb segment involving the cosmids FO2121 and AO4160, and more specifically the insertion's proximal breakpoint in the approximately 150-kb segment between FO2121 and FAT5 (PAX6), as the responsible factor for the patient's aniridia via a position effect resulting in functional haploinsufficiency of the PAX6 gene. This case illustrates the importance of recognizing that de novo complex chromosomal rearrangements found in patients with diverse clinical features may contribute to the phenotype, but that multiple mechanisms and higher levels of complexity may be unmasked by high resolution molecular cytogenetic studies. PMID- 16717456 TI - A 17q21.31 microdeletion encompassing the MAPT gene in a mentally impaired patient. AB - About 15% of patients with a clinical phenotype of Angelman syndrome (AS) have an unknown etiology. We report a patient with features reminiscent of AS, including a pattern of characteristic facial anomalies as well as speech impairment, developmental delay and frequent laughter. In addition, the patient had features not commonly associated with AS such as heart malformations and scoliosis. She was negative in SNURF-SNRPN exon 1 methylation studies and the G-banded karyotype was normal. Array-based comparative genomic hybridization disclosed a deletion of maximally 1 Mb at 17q21.31. The deleted region contains the MAPT gene, implicated in late onset neurodegenerative disorders, and the STH and NP_056258.1 genes. Another gene, such as CRHR1, might also be included based on maximum possible size of the deletion. We suggest that microdeletions within the 17q21.31 segment should be considered as a possible cause of phenotypes resembling AS, particularly when easily controlled seizures and/or cardiac abnormalities are also present. PMID- 16717457 TI - Comparative mapping of oculocutaneous albinism type II (OCA2), transient receptor potential cation channel, subfamily M member 1 (TRPM1) and two equine microsatellites, ASB08 and 1CA43, among four equid species by fluorescence in situ hybridization. PMID- 16717458 TI - Chromosomal localization, spatio-temporal distribution and polymorphism of the porcine tripartite motif-containing 55 (TRIM55) gene. PMID- 16717459 TI - Assignment of the oviductal glycoprotein 1 gene (OVGP1) to porcine chromosome 4q22-->q23 by radiation hybrid panel mapping. PMID- 16717460 TI - Assignment of the subunit C of succinate dehydrogenase complex (SDHC) gene to bovine chromosome 2 with somatic and radiation hybrid panel mapping. PMID- 16717461 TI - Assignment of the bovine PTGFR and HSD3B1 genes to bovine chromosome 3 with somatic and radiation hybrid panel mapping. PMID- 16717462 TI - Assignment of the proteolipid protein 1 gene (PLP1) to tammar wallaby chromosome Xq by fluorescence in situ hybridization with a BAC clone. PMID- 16717463 TI - Physical mapping of the ATP2A2 gene to equine chromosome 8p14-->p12 by FISH and confirmation by linkage and RH mapping. PMID- 16717464 TI - Physical mapping of immunoglobulin loci IGH@, IGK@, and IGL@ in the opossum (Monodelphis domestica). PMID- 16717465 TI - Impact of early tirofiban administration on myocardial salvage in patients with acute myocardial infarction undergoing infarct-related artery stenting. AB - BACKGROUND/AIMS: The timing of GpIIb/IIIa inhibitor administration may be important in achieving early epicardial and myocardial reperfusion. We evaluated the effect of early tirofiban on myocardial salvage and cardiovascular outcome in patients with acute myocardial infarction (AMI) undergoing infarct-related artery stenting. METHODS: Patients (n = 66) with a first AMI presenting <6 h from onset of symptoms were randomized to either early administration of tirofiban in the emergency room (n = 32) or later administration in the catheterization laboratory (n = 34) (tirofiban bolus dose of 10 microg/kg, followed by 0.15 microg/kg for 24 h). The primary end-point was the degree of myocardial salvage, determined by means of serial scintigraphic studies with technetium-99m sestamibi. Thirty-day major adverse cardiac events were also assessed. RESULTS: There were no significant differences in patient characteristics or in their presentation. The mean door-to-balloon time was similar in both groups (43 +/- 12 and 53 +/- 9 min, p = 0.08). The early and late treatment groups received tirofiban 18 +/- 4 and 52 +/- 10 min after admission, respectively. Angiographic analysis revealed a higher initial frequency of TIMI grade 3 flow in the early group (31% vs. 12%, p = 0.04). Procedural success was achieved in all patients. Myocardial risk area were comparable between early and late treatment groups (35.6 +/- 12.2% vs. 39.3 +/- 14.0%, p = 0.6). Scintigraphic outcomes demonstrated a significant reduction in the final infarction size (11.8 +/- 5.2% vs. 22.4 +/- 6.2%, p = 0.01), and improvement in salvage index (0.68 +/- 0.22 vs. 0.44 +/- 0.18, p = 0.003) in favor of the early tirofiban group. The thirty-day composite end-point of death, recurrent MI or rehospitalization also favored the early group (6% early, 15% late, p = 0.06). CONCLUSION: Early tirofiban administration enhanced the degree of myocardial salvage and clinical outcome in patients with AMI undergoing infarct-related artery stenting. PMID- 16717466 TI - Investigating the enhancing effect of music on autobiographical memory in mild Alzheimer's disease. AB - The enhancing effect of music on autobiographical memory recall in mild Alzheimer's disease individuals (n = 10; Mini-Mental State Examination score >17/30) and healthy elderly matched individuals (n = 10; Mini-Mental State Examination score 25-30) was investigated. Using a repeated-measures design, each participant was seen on two occasions: once in music condition (Vivaldi's 'Spring' movement from 'The Four Seasons') and once in silence condition, with order counterbalanced. Considerable improvement was found for Alzheimer individuals' recall on the Autobiographical Memory Interview in the music condition, with an interaction for condition by group (p < 0.005). There were no differences in terms of overall arousal using galvanic skin response recordings or attentional errors during the Sustained Attention to Response Task. A significant reduction in state anxiety was found on the State Trait Anxiety Inventory in the music condition (p < 0.001), suggesting anxiety reduction as a potential mechanism underlying the enhancing effect of music on autobiographical memory recall. PMID- 16717467 TI - Advantage of the right lateral position compared with the lithotomy position during the perineal procedure in abdominoperineal resection. AB - BACKGROUND/AIMS: Abdominoperineal resection (APR) is generally performed in the lithotomy position (LT-APR). The aim of this retrospective study was to evaluate the advantage of the right lateral (RL) position during the perineal phase of APR, in extended APR (ext-APR) that needed additional excision of the pelvic organs and sacrococcyx, and in total pelvic exenteration (TPE). METHODS: The present study is based on 50 patients observed from January 1993 to December 2004 (43 with primary rectal malignancy, 7 with recurrent cancer), who had undergone the following surgeries: LT-APR was carried out in 20 patients, RL-APR in 13, LT ext-APR in 5, RL-ext-APR in 8, LT-TPE in 2 and RL-TPE in 2. Perioperative factors were compared between the LT and RL position in each operative procedure. RESULTS: When RL-APR was compared with LT-APR, operative time (including the time to change the position) and the amount of operative blood loss were significantly less in RL-APR (mean +/- SE = 314 +/- 16 vs. 381 +/- 18 min, p = 0.0156 and 598 +/- 78 vs. 1,160 +/- 171 g, p = 0.0168, respectively). The blood loss and operative time were also significantly less in RL-ext-APR than in LT-ext-APR (1,060 +/- 170 vs. 3,590 +/- 1,270 g, p = 0.0277 and 353 +/- 31 vs. 488 +/- 41 min, p = 0.0219, respectively). The average blood loss and operative time were 4,190 g and 650 min in LT-TPE, and 1,450 g and 609 min in RL-TPE, respectively. CONCLUSION: The RL position during the perineal phase following the abdominal phase in the LT position significantly decreases blood loss and operative time in APR and ext-APR. PMID- 16717468 TI - Predicting anastomotic disruption after emergent small bowel surgery. AB - BACKGROUND/AIMS: Small bowel anastomoses performed in the emergent setting have a high risk of leakage. Attention to technical detail is imperative but does not guarantee success in these situations. We sought out factors that could play a role in the process of anastomotic dehiscence under these conditions. METHODS: 70 patients underwent 74 emergency small bowel anastomoses over a 21-month period in our institution during this prospective study. Patients with anastomotic disruption formed the case group and those without, the control group. Several preoperative, intraoperative and postoperative variables identified at the outset of the study were analyzed for possible associations with anastomotic dehiscence. RESULTS: Suture line disruption occurred in 26 of 74 anastomoses (35%). The duration of symptoms before presentation did not differ significantly between groups. Hypoalbuminemia (p = 0.004), hyponatremia at presentation (p = 0.012), and intraoperative hypotension (p = 0.042) were found to be significantly associated with disruption. Neither the nature of the primary pathology in the bowel nor the anastomotic level had a significant bearing on anastomotic leakage. CONCLUSION: Risk factors for leakage of emergent small bowel anastomoses include hypoalbuminemia, hyponatremia at presentation, and intraoperative hypotension. Under these circumstances, the creation of a temporary stoma or exteriorization may be a wiser option than primary anastomosis. PMID- 16717469 TI - Treatment of rectal cancer: reduction of local recurrence after the introduction of TME - experience from one University Hospital. AB - BACKGROUND: Local recurrence (LR) of cancer after rectal surgery is followed by significant morbidity and mortality. Since the introduction of total mesorectal excision (TME) the rates of LR have decreased in many centres. The aim of this retrospective study was to investigate the effect of TME on the recurrence rates of rectal cancer and the impact of the surgeons. METHODS: All patients resected for invasive rectal cancer from 1990 until 2000 were initially included in the study. From February 1994, TME was adopted as the standard treatment (TME group). Before this period, rectal surgery was performed by the non-TME technique (non TME group). To obtain homogeneity, patients who underwent preoperative irradiation, emergency operations, pre- or intraoperative bowel perforation, residual tumour stage (R1,2) including Dukes' D stage and postoperative mortality within 31 days, were excluded. 139 patients in the non-TME group and 181 patients in the TME group were found eligible for analyses. RESULTS: The estimated LR rate at 1, 3 and 5 years was 7, 15 and 17% (non-TME) versus 4, 9 and 9% (TME) (p = 0.046, log-rank test). The anastomotic leakage rate was 6% (non-TME) versus 4% (TME) (not significant). Perioperative blood loss >500 ml, reoperations during the hospital stay and lymph node (N) stage were the independent risk factors for LR in the multivariate analysis. The case volume did not significantly influence LR rates. However, the variability of individual surgical results was reduced after the introduction of TME. CONCLUSIONS: TME yields significantly lower LR rates compared with traditional surgery. Since the introduction of TME, experience with rectal surgery has been gathered by a limited number of surgeons. The results of individual surgeons have consistently improved and the variability of individual surgical results is now at a lower level. PMID- 16717470 TI - Postoperative joint symptoms: a risk factor for pouchitis in ulcerative colitis patients. AB - BACKGROUND: The prevalence and significance of joint symptoms appearing in patients with ulcerative colitis (UC) following a colectomy are unclear. AIM: We investigated the relationship between joint symptoms during steroid tapering following an ileal pouch-anal anastomosis (IPAA) and the cumulative risk for developing pouchitis. PATIENTS AND METHODS: The medical records of 571 patients who underwent an IPAA with a mucosectomy were retrospectively reviewed to evaluate their joint symptoms. A diagnosis of pouchitis was obtained using the Pouchitis Disease Activity Index (PDAI) and the cumulative risk of pouchitis was estimated using a Kaplan-Meier life table analysis. RESULTS: Joint symptoms during steroid tapering were reported by 126 (22.0%) of the UC patients and each of those had involvement of the small joints of the hand. The main symptoms were pain and stiffness, especially in the morning. The cumulative risk for developing pouchitis after 10 years was found to be 20% in patients who experienced joint symptoms during steroid tapering and 10% in those without those symptoms (p = 0.001). CONCLUSION: The presence of joint symptoms during steroid tapering is a significant risk factor for the development of pouchitis in patients who have undergone an IPAA for UC. PMID- 16717471 TI - Long-term graft outcome of pediatric liver transplantation in Copenhagen: analysis of the first 51 cases. AB - BACKGROUND: Graft loss after liver transplantation remains a significant problem, especially in pediatric patients. The aim of this study was to assess our initial series of pediatric liver transplantation and to identify the risk factors that influence graft outcome. METHODS: The first 51 transplantations were analyzed retrospectively. All transplantations were stratified into three groups according to graft type (full-size, reduced-size, and living-related-donor graft). Survival data of the grafts were stratified and multivariate analysis conducted with respect to preoperative and surgical factors. RESULTS: Seventeen of all the transplants were full-size grafts and 34 technical-variant grafts (27 reduced size grafts from cadavers and 7 living-related-donor grafts). The overall graft survival rates were 65, 62 and 53% at 1, 3 and 5 years, respectively. Twenty three of 51 grafts (45%) were lost. Poor status of the recipients (hospitalization or intensive care unit care before surgery), a retransplanted graft, and a reduced-size graft were independent risk factors for graft failure. With experience, overall graft survival has improved significantly and the differences in graft survival between graft types have disappeared. CONCLUSIONS: To improve graft survival after pediatric liver transplantation, the timely referral of potential recipients to the transplant team and employing a meticulous technique during the operation, particularly for the technical-variant graft, are required. PMID- 16717472 TI - Epidermal growth factor receptor expression in pancreatic carcinoma using tissue microarray technique. AB - BACKGROUND: The effect of overexpression of epidermal growth factor receptor (EGFR) in pancreatic carcinoma is not clear. Utilizing tissue microarrays, we evaluated EGFR expression in pancreatic cancer to determine the association of EGFR expression with histopathologic characteristics and patient outcome. METHODS: 71 cases of pancreatic adenocarcinoma and 18 cases of chronic pancreatitis were retrieved from archival files. Tissue cores from donor blocks were arrayed to create a tissue microarray. Sections were stained with EGFR and the intensity of membranous staining and percentage of tumor cells showing immunoreactivity were determined. At least 1% membranous staining and 1+ intensity were considered positive. RESULTS: EGFR was present in 49 of 71 (69%) cases of pancreatic adenocarcinoma and 7 of 18 (39%) cases of chronic pancreatitis (p = 0.03). There was no statistically significant correlation between intensity or extent of EGFR expression and tumor grade, size, or lymph node status. While median survival was nearly twice as long when EGFR was expressed (15.2 vs. 8.3 months), this did not reach statistical significance. CONCLUSIONS: EGFR is overexpressed in pancreatic cancers independent of histopathologic characteristics and does not predict survival. Immunohistochemical analysis of EGFR staining may help in identifying candidates for EGFR inhibitor therapy. PMID- 16717473 TI - Compartimentalization for chylothorax originating from the abdomen after extended esophagectomy. Report of two cases and review of the literature. AB - BACKGROUND: Chyle leakage from the chest after extended esophagectomy originating from the abdomen is a rare complication with various clinical presentations and treatments. METHODS: Two cases of chylothorax originating from the abdomen are discussed and the literature concerning diagnosis, management and outcome is reviewed. RESULTS AND CONCLUSION: Initially conservative measures should be installed; however, prolonged conservative treatment should be avoided. Reoperation gives an opportunity to identify the leak. If the leakage originates from the abdomen, compartimentalization is the essential step to solve the problem. PMID- 16717474 TI - Prevalence and risk factors of HPV infection among high-risk rural and urban Lithuanian women. AB - AIMS: To investigate the prevalence, persistence and risk factors of high oncogenic risk human papillomavirus (HPV) among urban and rural women of reproductive age coming to consult a gynaecologist. METHODS: A prospective cohort study in urban (Kaunas) and rural (Marijampole) regions of Lithuania. The data were collected in 8 healthcare institutions from women seeking consultation of gynaecologists using a questionnaire for finding out demographic, social, behavioural and biomedical factors. HPV DNA was determined by molecular hybridization method (hybrid capture version II) determining HPV of high oncogenic risk. RESULT: 1,120 women participated in the study. The prevalence of high-risk HPV among the studied women was 25.1%. It was higher among the urban women than among the rural women. The prevalence of high-risk HPV was increased if the subjects had 2 or more sexual partners during the last 12 months (OR 2.81; 95% CI 1.83-4.32), were 19 years of age or younger (OR 2.68; 95% CI 1.47-4.91), were smoking (OR 1.81; 95% CI 1.16-2.81), and had secondary or lower education level (OR 1.43; 95% CI 1.01-2.04). This infection was obviously associated with high- and low-grade squamous intraepithelial changes of the cervix (OR 1.66, 95% CI = 1.08-2.53). CONCLUSION: The incidence rate for cervical cancer in Lithuania is one of the highest in comparison with other European countries. HPV infection was also particularly common in the studied population. About one-fourth of the women were infected with high-risk HPV infection. Young and less educated women were found to be the group that was most exposed to HPV, and therefore public health interventions and education seem to be essential in programs aimed at reducing the incidence of cervical cancer. PMID- 16717475 TI - Estimating haplotype effects on dichotomous outcome for unphased genotype data using a weighted penalized log-likelihood approach. AB - OBJECTIVE: To develop a method to estimate haplotype effects on dichotomous outcomes when phase is unknown, that can also estimate reliable effects of rare haplotypes. METHODS: In short, the method uses a logistic regression approach, with weights attached to all possible haplotype combinations of an individual. An EM-algorithm was used: in the E-step the weights are estimated, and the M-step consists of maximizing the joint log-likelihood. When rare haplotypes were present, a penalty function was introduced. We compared four different penalties. To investigate statistical properties of our method, we performed a simulation study for different scenarios. The evaluation criteria are the mean bias of the parameter estimates, the root of the mean squared error, the coverage probability, power, Type I error rate and the false discovery rate. RESULTS: For the unpenalized approach, mean bias was small, coverage probabilities were approximately 95%, power ranged from 15.2 to 44.7% depending on haplotype frequency, and Type I error rate was around 5%. All penalty functions reduced the standard errors of the rare haplotypes, but introduced bias. This trade-off decreased power. CONCLUSION: The unpenalized weighted log-likelihood approach performs well. A penalty function can help to estimate an effect for rare haplotypes. PMID- 16717476 TI - Association between alcohol intake and domain-specific cognitive function in older women. AB - Moderate levels of alcohol intake may be associated with better cognitive function; however, this relationship may vary between cognitive domains. Women, aged 65-80 years, enrolled in the Women's Health Initiative (WHI) randomized clinical trials of hormone therapy, underwent annual standardized testing for global cognitive function through the ancillary WHI Memory Study (average follow up of 4.5 years) and domain-specific cognitive function through the WHI Study of Cognitive Aging (average follow-up of 1.7 years). Compared to nondrinkers, women reporting moderate levels of alcohol intake (or=1 drink/day by 5.7% for verbal knowledge (p < 0.001) and by 5.7% for phonemic fluency (p = 0.004), compared to never-drinkers. Moderate levels of alcohol intake are associated with somewhat better cognition, which may be expressed most strongly in functions related to verbal knowledge and phonemic fluency. However, our observational study cannot rule out confounding associations with unmeasured factors. PMID- 16717477 TI - Effects of itopride hydrochloride on plasma gut-regulatory peptide and stress related hormone levels in healthy human subjects. AB - Itopride hydrochloride (itopride), a gastrokinetic drug, has recently been evaluated for its clinical usefulness in functional dyspepsia. We investigated effects of itopride on human plasma gastrin-, somatostatin-, motilin-, and cholecystokinin (CCK)-like immunoreactive substances (IS); adrenocorticotropic hormone (ACTH)-immunoreactive substances (IS), and cortisol under stress conditions in healthy subjects. A single administration of itopride caused significant increases in plasma somatostatin- and motilin-IS levels compared to placebo. Itopride significantly decreased plasma CCK-IS, and suppressed the ACTH IS level compared to placebo. We hypothesize that itopride may have an accelerating gastric emptying effect, and a modulatory effect on the hypothalamo pituitary-adrenal axis and autonomic nervous functions. These effects might be beneficial in stress-related diseases, suggesting that itopride has clinicopharmacological activities. PMID- 16717478 TI - Role of NO and prostaglandins in acute hypoxic vasoconstriction in sheep pulmonary veins. AB - The aim of this study was to investigate the effect of hypoxia on and the role of nitric oxide (NO) and cyclooxgenase inhibition in hypoxia-induced vasoconstriction in sheep isolated pulmonary veins. We used the potent pulmonary vasoconstrictor U46619, a thromboxane analog, as a precontractile agent. Our results showed that hypoxia caused a vasoconstriction both under resting tone and in U46619 (10(-6) mol/l) precontracted pulmonary veins. In the presence of the nonselective NO synthase inhibitior Nomega-nitro-L-arginine methyl ester (L-NAME; 3 x 10(-5) mol/l), the hypoxic pulmonary vasoconstriction (HPV) was significantly increased in veins under resting force. However, there was a decrease in HPV in pulmonary veins precontracted with U46619 in the presence of L-NAME. Moreover, L NAME markedly augmented the U46619-induced pulmonary contractions under normoxic conditions. Cyclooxygenase inhibition with indomethacin (10(-5) mol/l) significantly reduced the HPV both under resting tone and in precontracted veins. Indomethacin also significantly decreased the U46619-induced pulmonary contractions prior to the induction of hypoxia. Our findings suggest that NO and prostaglandins can act as a modulators of the hypoxic vasoconstriction in isolated pulmonary veins. PMID- 16717479 TI - Full and partial post-traumatic stress disorder among World War II prisoners of war. AB - BACKGROUND: The aim of the study is to evaluate the frequency and characteristics of full and partial post-traumatic stress disorder (PTSD) among 66 World War II Nazi prisoners of war, some of whom had been deported to death camps. METHODS: The Structured Clinical Interview for DSM-IV was used to assess the presence of PTSD and major depression. The Dissociative Experiences Scale and the 58-item version of the Hopkins Symptom Checklist were also administered. Partial PTSD was defined as the presence of at least one symptom in each of the clusters defined by DSM-IV. RESULTS: Forty-eight percent of the subjects have a partial PTSD, while 20% reported the full syndrome. When compared to subjects with full PTSD, those with partial PTSD showed a significantly lower frequency of comorbid depression and lower levels of psychological distress, as measured by the Symptom Checklist. No differences were found with regards to dissociative symptoms and help-seeking. CONCLUSIONS: The problem of definition of PTSD partial syndromes deserves more attention in the literature. PMID- 16717480 TI - Narcissistic regulation of the self and interpersonal problems in depersonalized patients. AB - BACKGROUND: Psychoanalytical theories coincide in understanding depersonalization (DP) as a disorder of narcissistic self-regulation. DP is described as an ego defense against overwhelming shame resulting in a splitting of an observing ego detached from the experiencing self. In contrast to a behavioral-cognitive theory on DP, which suggests that the catastrophic appraisal of normal transient DP maintains the disorder, psychodynamic approaches stress that DP is an important defensive function for the individual. We examine this psychodynamic aspect more closely as it relates to narcissistic self-regulation and interpersonal behavior in depersonalized patients. SAMPLING AND METHODS: Thirty-five patients with pathological DP are compared with 28 patient controls concerning their narcissistic self-regulation and interpersonal behavior. For the assessment, we used the German Narcissism Inventory and the Inventory of Interpersonal Problems. The two groups were controlled for sociodemographic data, comorbidity with a personality disorder, and the General Severity Index of the Symptom Check List-90 R. RESULTS: Bonferroni-corrected group comparison showed that the depersonalized patients are characterized by perceiving themselves as helpless, hopeless, socially isolated and worthless, perceiving others as bad and disappointing, and that they avoid interpersonal relations and reality significantly more than other patients with equal symptom severity. CONCLUSIONS: Treatment approaches on DP should take the issue of low self-esteem, pervasive shame and the related defensive social avoidance into account. Further empirical research on psychodynamic concepts of DP is warranted also for the sake of linking modern neurobiological findings with clinical experience. PMID- 16717481 TI - Auditory recognition memory, conscious recollection, and executive function in patients with schizophrenia. AB - BACKGROUND: Dual-process models propose that recognition memory (RM) involves two processes: conscious recollection and familiarity-aware memory. Studies investigating RM in schizophrenia report a selective deficit in conscious recollection and intact levels of familiarity-driven RM for stimuli presented in the visual and olfactory domains. It has been suggested that abnormalities in conscious recollection result from a breakdown in frontal strategic memory processes involved in encoding and retrieval and executive functions linked to reality monitoring and decision making. We investigated three predictions arising from these proposals. Firstly, if conscious recollection abnormalities arise from a central impairment, then these abnormalities should not be domain specific. Secondly, if the deficits in conscious recollection arise from a breakdown in executive processes, deficiencies should be correlated with executive dysfunction. Finally, the conscious recollection deficiencies are likely to be more severe in schizophrenia, a condition associated with marked executive dysfunction relative to Major Depressive Disorder, Recurrent (MDDR), in which executive dysfunction is less marked. METHODS: The remember/know paradigm was used to investigate RM for voices in three groups: patients with schizophrenia (n = 14), patients with MDDR (n = 16), and normal controls (n = 16). Executive function was assessed using the Wisconsin Card Sorting Task. RESULTS: Patients with schizophrenia made significantly fewer remember responses than normal controls (p < 0.01), despite normal levels of discrimination and familiarity driven auditory RM. Patients with MDDR did not differ significantly from either normal controls or patients with schizophrenia. Executive dysfunction was limited to the schizophrenia group and was not correlated with conscious recollection deficiencies. CONCLUSIONS: Patients with schizophrenia exhibit a deficit in conscious recollection for auditory RM of voices. These findings, when considered alongside remember/know data collected from the same set of patients for olfactory and visual RM, support proposals that abnormalities in conscious recollection stem from a breakdown in central rather than domain-specific processes. PMID- 16717483 TI - Japanese patients' descriptions of "the good nurse": personal involvement and professionalism. AB - According to recent nursing literature, virtue ethics is regaining attention as a way to establish a comprehensive nursing ethical model, which will appropriately reflect actual nursing. This study explored the characteristics of "the good nurse" as an ethical ideal from Japanese patients' perspectives. The findings described the good nurse for Japanese nursing care recipients and delineated their expectations of her or him. For Japanese patients, the good nurse was a person with personal involvement and professional comportment. The study revealed the participants' perspectives of the importance of person-to-person relationships between nurses and patients, and of nurses' professional disposition and competency. PMID- 16717482 TI - A qualitative analysis of the meaning of aging for women with disabilities with policy implications. AB - This is a report of a hermeneutic phenomenological study of the meaning of aging for women with childhood-onset disabilities due to the effects of paralytic polio. Twenty-five women aged 55-65 years were interviewed 2 to 4 times regarding their life course experiences and the meaning they assigned to aging. Field notes, audiotaped interviews, life course charts, and demographics were used in thethematic analysis that produced 5 themes: Bodies Change, Disrupted Meaning, The Unpredictibility of Aging, Slowing Down, and Changing Perspective. Overall, the findings indicate that the experiences of disability due to the result of paralytic polio could not be separated from the experiences of aging, which ultimately led the author to question policies that distribute and fund benefits based on age and disability status without an understanding of the varied experiences of women with disabilities. PMID- 16717484 TI - Bringing safety and responsiveness into the forefront of care for pregnant and parenting aboriginal people. AB - Poor access to prenatal care for Aboriginal people is well documented, and is explicated as an unethical barrier to care resulting from colonial and neocolonial values, attitudes, and practices. A postcolonial standpoint, participatory research principles, and a case study design were used to investigate 2 Aboriginal organizations' experiences improving care for pregnant and parenting Aboriginal people. Data were collected through exploratory interviews and small-group discussions with purposefully selected community leaders, providers, and community members. The study found that safety in healthcare relationships and settings, and responsiveness to individuals' and families' unique experiences and capacities must be brought into the forefront of care. Results suggest that the intention of care must be situated within a broader view of colonizing relations to improve early access to, and relevance of, care during pregnancy and parenting for Aboriginal people. PMID- 16717485 TI - Mutual moral caring actions: a framework for community nursing practice. AB - A community practice framework is presented as the synthesis of research findings from the analysis of a critical ethnonursing study of women in recovery from chemical dependence. Critical Social Theory is used to examine the paradoxical experiences of women from their lifeworld and system within the community. The framework focuses on the mutual moral caring actions of the community nurse and the women in the recovery. It is supported by the concepts of transcultural nursing ethics. The utility of the framework is to promote clarity of speech and parity of community membership for women in recovery from chemical dependence and their return to the community. PMID- 16717487 TI - Results from the Nurse Manifest 2003 study: nurses' perspectives on nursing. AB - The purpose of this article is to raise awareness, inspire action, and open dialogue about professional values and workplace conditions. Results of the Nurse Manifest 2003 Study are presented through meta-synthesis of group summaries answering the questions: What is it like to practice nursing? and, What changes do nurses desire to support practice? The results illuminate the underlying framework of professional values that splits our profession, nursing units, and educational institutions. An analysis of the similarities and differences within and between groups reflect values which are rooted in the history of our profession and affected by present working conditions. PMID- 16717486 TI - Taking responsibility: toward an understanding of morality in practice: a critical review of the empirical and selected philosophical literature on the social organization of responsibility. AB - Although the nursing literature overflows with references to the myriad things for which nurses and patients are de facto responsible, nurses have never explicitly examined the social construction of responsibility in any clinical context. This article reviews and integrates the empirical and philosophical literature on moral responsibility in the context of mental health nursing. It selectively reviews both traditional and feminist philosophical accounts to more deeply understand the socially constructed nature of responsibility and the implications for understanding morality in practice. It seeks to illuminate the concept of "taking responsibility" qua moral responsibility and asks what makes this notion of responsibility particularly "moral." PMID- 16717489 TI - An ethic of analysis: an argument for critical analysis of research interviews as an ethical practice. AB - Nursing literature is replete with discussions about the ethics of research interviews. These largely involve questions of method, and how careful study design and data collection technique can render studies more ethical. Analysis, the perennial black box of the research process, is rarely discussed as an ethical practice. In this paper, I introduce the idea that analysis itself is an ethical practice. Specifically, I argue that political discourse analysis of research interviews is an ethical practice. I use examples from my own research in a prison control unit to illustrate what this might look like, and what is at stake. PMID- 16717490 TI - Moving beyond the "exotic": applying postcolonial theory in health research. AB - Postcolonialism offers nursing scholarship a framework for understanding culture and identity as fluid and complex, historically situated, and discursively constructed. This article describes one version of implementing postcolonial theory, using examples from a research project conducted with urban American Indians on the topic of diabetes. I demonstrate the influence and value of postcolonialism throughout the research process. A postcolonial approach can help nursing researchers and practitioners avoid reproducing injustices and stereotypes, illuminate the complexities of life at the intersections, and contribute to the construction of a more socially just world. PMID- 16717492 TI - Fundamental or foundational obligation? Problematizing the ethical call to spiritual care in nursing. AB - Spiritual nursing care is increasingly being cited in the nursing literature as a fundamental ethical obligation. This obligation is based upon the argument that nurses provide holistic care, spirituality is a universal dimension of the person, and so nurses should care for the spiritual dimension. However, the literature on the spiritual dimension in nursing illustrates widely differing foundational assumptions about this important aspect of care. The philosophic categories of humanism, theism, and monism can be used to illustrate the different understandings of the spiritual dimension, and the implications of these understandings for the competence of the nurse and the nature of the nurse patient interaction in the context of spiritual care. PMID- 16717491 TI - Ethical multiculturalism: an evolutionary concept analysis. AB - A concept analysis of ethical multiculturalism using Rogers' evolutionary approach is presented. It includes attributes, antecedents, and consequences of ethical multiculturalism. Attributes include moral reasoning, cultural competence, beneficence/nonmaleficence, and respect for persons and communities. Antecedents are culture knowledge; cultural awareness, sensitivity, encounters, and skill; and understanding of ethical principles. Consequences include protection of human subjects, preservation of cultural norms, dignity of participants and communities, and perceived value by individuals and communities. Ethical multiculturalism is defined as the use of moral reasoning to apply the ethical principles of beneficence and respect for persons and communities in a culturally competent manner to research in various societies or cultures. PMID- 16717493 TI - Better reproductive healthcare for women with disabilities: a role for nursing leadership. AB - This paper examines the reproductive healthcare experiences of women with disabilities in the light of commonly accepted principles of biomedical ethics. Recommendations are made for nursing to assume a leadership role in reducing gender and disability inequity in health care. PMID- 16717494 TI - Listening in thin places: ethics in the care of persons with Alzheimer's disease. AB - Healthcare professionals may see prolonging life and use of scarce resources as futile if the quality of life of persons with Alzheimer's disease is deemed to be poor. Thus, nurses and other healthcare providers need to question traditional assumptions of quality of life. This article presents a phenomenological study of ethical concerns related to quality of life as described by persons with Alzheimer's disease, family caregivers, and professional caregivers. The theme of "Creating a Quality of Life Through Connected Lives" calls forth an ethics of respect for the individual experience and its connecting relationships. PMID- 16717495 TI - An ethics of testimony: prisoner nurses at Auschwitz. AB - This historical research report presents and analyzes 2 recently identified narratives of women who underwent sterilization experiments at the Auschwitz concentration camp during World War II. A description of the historical and contextual background is presented in which involvement of the prisoner nurse occurred in the sterilization experiments. Using a critical feminist perspective, the ethics of nursing involvement are discussed in these experiments, with an emphasis on the political dimension. Salient implications are explored for contemporary nursing. PMID- 16717496 TI - Mobile subjectivities: positioning the nonunitary self in critical feminist and postmodern research. AB - Most scholarly work is written from the perspective of the author being a unitary subject occupying a sole, rational, and unified position. This article argues that scholarship may be enhanced by the author adopting multiple subject positions as a methodological framework. Such an adoption is advantageous in working against the romance of the notion of a single truth while also maintaining teleological values congruent with critical and feminist agendas. This article outlines the conceptual development of this methodological framework, the rationale for its development, an explication of the concept of multiple subjectivity, and an exemplar of its application within nursing research. PMID- 16717497 TI - Bridging the distance between the objectivism of research and the subjectivity of the researcher. AB - A phenomenological study of researchers' experience of their work is presented. The disparity between researchers' subjective experience and the inherent objectivism of science and research is discussed. Bridging the distance between the immediate experience of research and the positivistic/objectivistic nature of research is proposed as a way to keep the research community cognizant of values and beliefs that may get pushed aside in the clamor for factual information and material gain. A synthesis of intentionality is discussed as a method for disclosing a researcher's constitutive part in phenomenological research. Expanding the traditional format for research reports to include a brief biographical account of the inception of the research is proposed and an example included. PMID- 16717498 TI - Insomnia in patients with traumatic brain injury: frequency, characteristics, and risk factors. AB - OBJECTIVES: To document the frequency of insomnia (according to DSM-IV and ICSD criteria), to describe its sociodemographic and clinical characteristics, and to identify potential predictors of insomnia in persons with traumatic brain injury (TBI). PARTICIPANTS AND PROCEDURE: Four hundred fifty-two participants aged 16 years and older with minor to severe TBI answered a questionnaire pertaining to quality of sleep and fatigue. MAIN OUTCOME MEASURES: Proportion of participants fulfilling criteria for insomnia symptoms or syndrome. Validated measures of insomnia severity, fatigue level, and psychologic distress. Results of a logistic regression analysis. RESULTS: Overall, 50.2% of the sample reported insomnia symptoms and 29.4% fulfilled the diagnostic criteria for an insomnia syndrome. For the latter participants, insomnia was a severe and chronic condition remaining untreated in almost 60% of cases. Risk factors associated with insomnia were milder TBIs, and higher levels of fatigue, depression, and pain. CONCLUSION: Insomnia is a prevalent condition after TBI requiring more clinical and scientific attention as it may have important repercussions on rehabilitation. PMID- 16717499 TI - Outcomes in TBI with violent versus nonviolent etiology in a predominantly rural setting. AB - OBJECTIVE: To evaluate differences in outcome in persons with violent versus nonviolent traumatic brain injury (TBI) etiology. DESIGN: Two-group (violent vs nonviolent) 1-year follow-up study. SETTING: Midwestern medical center TBI Model System serving a predominantly rural catchment area. PARTICIPANTS: Forty-five (n = 19 violent TBI etiology; n = 26 nonviolent TBI etiology) inpatients with primary diagnosis of TBI followed up as outpatients 1 year after injury. MAIN OUTCOME MEASURES: Substance use, income source, employment status, Wechsler Adult Intelligence Scale--Revised (abbreviated version), Logical Memory I and II from the Wechsler Memory Scale--Revised, Wide Range Achievement Test--Third Edition Reading subtest, Trail-Making Tests A and B, Rey Auditory Verbal Learning Test, Community Integration Questionnaire, Neurobehavioral Functioning Inventory. RESULTS: Members of the violent group were more likely to be men, of a racial minority, unemployed, and have low income. Substance abuse was common among both groups prior to injury, with significant declines at 1 year. Follow-up also revealed significant group differences in verbal intelligence, social integration, productivity, and source of income, but nonsignificant difference in employment. In all comparisons, more favorable outcomes were found for the nonviolent group. CONCLUSIONS: Persons with violent injury etiology have poorer premorbid functioning and are likely to have less favorable outcomes than the general population with TBI. Therefore, persons with violent TBI etiology may require more intensive aftercare programming to promote improved rehabilitation outcomes. PMID- 16717500 TI - Impaired awareness of deficits and treatment adherence among people with traumatic brain injury or spinal cord injury. AB - OBJECTIVE: To examine the relation between impaired awareness of deficits (IAD) and treatment adherence and to verify previous findings regarding the types of disabilities that people with traumatic brain injury (TBI) tend to underestimate. DESIGN: Cross-sectional study. PARTICIPANTS: Twenty-four persons with moderate to severe TBI and 16 persons with traumatic spinal cord injury (SCI) admitted to an inpatient neurorehabilitation program. MAIN OUTCOME MEASURES: IAD assessed using the short version of the Problem Checklist of the Head Injury Family Interview and treatment adherence using the Medical Regimen Adherence Scale. RESULTS: Presence of IAD is linked with poor adherence. Patients with TBI significantly underestimate their emotional/behavioural and cognitive disabilities, but accurately assess their physical disabilities, whereas patients with SCI accurately assess all spheres. Patients with TBI are significantly less aware of their disabilities compared with the patients with SCI as expected. CONCLUSIONS: These results confirm IAD's link to treatment adherence, and support the observation that IAD is a common clinical feature of brain injury that should be carefully considered when making prognoses and developing and applying interventions with this population. PMID- 16717501 TI - Predicting family functioning after TBI: impact of neurobehavioral factors. AB - OBJECTIVE: To identify risk factors for poor family functioning and neurobehavioral problems after traumatic brain injury (TBI) or orthopedic injuries (OI). DESIGN: Longitudinal analyses of data from an inception cohort. PARTICIPANTS: Seventy-five patients with moderate/severe TBI, 47 patients with mild TBI, and 44 patients with OI at discharge; and 49 patients with moderate/severe TBI, 24 patients with mild TBI, and 33 patients with OI at 1-year follow-up. OUTCOME MEASURES: Measures of family functioning (Family Assessment Device) and Neurobehavioral Functioning Index at hospital discharge and 1-year follow-up. RESULTS: At discharge, patients with moderate/severe TBI had more symptoms of depression, memory/attention problems, and motor impairments than patients with OI and greater communication difficulties than patients with OI or mild TBI. At follow-up, patients with moderate/severe TBI continued to have more problems in memory/attention, depression, and communication. Approximately one third of each group had unhealthy family functioning at each assessment period. Few patients reported both impaired family functioning and clinical depression. Distressed family functioning correlated strongly with increased rates of neurobehavioral symptoms. Family dysfunction at follow-up was best predicted by family dysfunction at discharge and depression or memory/attention deficits at follow-up. CONCLUSIONS: After TBI, patients at the greatest risk for distress at follow-up were those with family dysfunction at discharge and continued neurobehavioral problems. High-risk families need to be identified so that necessary referrals and/or treatment can be offered. PMID- 16717502 TI - Coping and emotional adjustment following traumatic brain injury. AB - OBJECTIVES: To examine the association between coping style and emotional adjustment following traumatic brain injury. PARTICIPANTS: Thirty three individuals who had sustained a traumatic brain injury (mean duration of posttraumatic amnesia = 32 days) between 1(1/2) months and almost 7 years previously. MEASURES: Coping Scale for Adults, Hospital Anxiety and Depression Scale, Rosenberg Self-Esteem Scale, State-Trait Anger Expression Inventory, and the Sickness Impact Profile. RESULTS: Approximately 50% of the sample reported clinically significant levels of anxiety and depression. Coping characterized by avoidance, worry, wishful thinking, self-blame, and using drugs and alcohol was associated with higher levels of anxiety, depression, and psychosocial dysfunction and lower levels of self-esteem. Coping characterized by actively working on the problem and using humor and enjoyable activities to manage stress was associated with higher self-esteem. Lower premorbid intelligence (measured via the National Adult Reading Test) and greater self-awareness (measured via the Self-Awareness of Deficits Interview) were associated with an increased rate of maladaptive coping. CONCLUSIONS: The strong association between the style of coping used to manage stress and emotional adjustment suggests the possibility that emotional adjustment might be improved by the facilitation of more adaptive coping styles. It is also possible that improving emotional adjustment may increase adaptive coping. The development and evaluation of interventions aimed at facilitating adaptive coping and decreasing emotional distress represent important and potentially fruitful contributions to enhancing long-term outcome following brain injury. PMID- 16717504 TI - Usefulness of diffusion tensor imaging for evaluation of motor function in patients with traumatic brain injury: three case studies. AB - OBJECTIVES: To determine whether diffusion tensor imaging (DTI) can detect diffuse axonal injury, and to evaluate the association of DTI findings with motor function in patients with traumatic brain injury. DESIGN: Three case studies. SETTING: An inpatient rehabilitation unit in Korea. PARTICIPANTS: Three patients with traumatic brain injury in whom conventional neuroimaging showed normal appearing white matter. MAIN OUTCOME MEASURES: Patients were studied with DTI. Fractional anisotropy (FA) was measured from 3 different anatomic locations on both sides of the corticospinal tract. Motor function was evaluated using the motoricity index. RESULTS: Fractional anisotropy tended to be reduced in normal appearing corticospinal tracts that were remote from the involved segment. Diffusion tensor imaging showed reduction of FA in areas consistent with motor dysfunction. CONCLUSION: Fractional anisotropy of the corticospinal tract may be used in the detection of diffuse axonal injury. The association between decreased motoricity index and decreased FA suggests that DTI may be useful in evaluating patients with traumatic brain injury. PMID- 16717503 TI - Complicated mild traumatic brain injury on the inpatient rehabilitation unit: a multicenter analysis. AB - OBJECTIVE: To determine characteristics of patients with complicated mild traumatic brain injury (CMTBI) on the inpatient rehabilitation unit and to accentuate limits of current classification systems for patients with mild TBI. DESIGN: Multicenter analysis of individuals with a Glasgow Coma Scale (GCS) score of 13 to 15 (lowest score in 24 h) and positive computed tomography findings admitted to inpatient rehabilitation for TBI. SETTING: 16 TBI Model System centers funded by the National Institute on Disability and Rehabilitation Research. PARTICIPANTS: 373 adults with CMTBI. MAIN OUTCOME MEASURES: Functional Independence Measure (FIM) and Disability Rating Scale (DRS) scores, length of stay, and medical complications. RESULTS: 16% of the sample experienced serious cranial complications (eg, cerebrospinal fluid leak, intracranial hypertension), with seizures being the most common during the course of inpatient rehabilitation. Most patients demonstrated substantial improvements on functional outcome measures during rehabilitation (mean admission FIM 68.2, discharge FIM 100.6; admission DRS 9.1, discharge DRS 4.8). Age, acute length of stay, and FIM score on rehabilitation admission were significant predictors of rehabilitation length of stay. Factors not predictive of rehabilitation duration were GCS score, ethnicity, gender, incidence of fractures, and incidence of cranial/noncranial complications. CONCLUSION: Despite favorable GCS scores, some patients with CMTBI suffer a variety of serious cranial and noncranial complications. Such patients appear to benefit from additional brain injury services offered in inpatient rehabilitation. Further study of the unique treatment needs of these patients is warranted. PMID- 16717505 TI - Gender, sex steroids, and neuroprotection following traumatic brain injury. PMID- 16717506 TI - Update on antidepressants. PMID- 16717507 TI - Corticosteroids in TBI: is the story closed? PMID- 16717511 TI - Pouch design and size paradox (aka) the potty paradox-a topic for an "incontinent" ostomy consensus panel. PMID- 16717512 TI - Barriers to incontinence research funding. PMID- 16717513 TI - Reporting P values. PMID- 16717514 TI - Pressure ulcer pain. AB - OBJECTIVE: To describe and interpret the complexities of the pain experienced by persons with painful pressure ulcers. DESIGN: Heideggerian hermeneutic phenomenology. SETTING AND SUBJECTS: Ten participants with pressure ulcers hospitalized in an acute-care community hospital were interviewed. METHODS: Multiple taped interviews, transcripts, and field notes were collected over an 8 month period. Data analysis occurred over a 16-month period and included an initial process of naming the text, identifying themes, and writing the interpretations of each participant's story. Secondary analysis comprised reading across the texts for commonalities and shared meanings. These data were related to other literature sources, shared with a team of researchers familiar with the Heideggerian hermeneutic phenomenology process, and with wound, ostomy, and continence nurses. RESULTS: A total of 22 themes and 1 constitutive pattern were identified. Participants with varying stages of pressure ulcers shared poignant stories about their pain. Many related their accounts and instances of actions by health personnel and the treatment interventions that were painful. Some related their accounts of communication failures that contributed to stress, tension, and anxiety. CONCLUSIONS: The findings of this study clearly indicate the need for increased attention to pain assessment and pain management in nursing education, nursing practice, and nursing research. The findings also suggest that the commonly used definition of pain is insufficient and needs revision. PMID- 16717515 TI - EPUAP pressure ulcer prevalence survey in Sweden: a two-year follow-up of quality indicators. AB - OBJECTIVE: The aim of this study was to determine the effects of quality improvement programs for pressure ulcer prevention by conducting a follow-up survey in a hospital in Sweden. DESIGN: A cross-sectional survey design with comparison between data collected in 2002 and 2004. SETTING AND SUBJECTS: All inpatient areas were surveyed in the surgical, medical, and geriatric departments in a university hospital. A total of 369 patients were included. INSTRUMENTS: The European Pressure Ulcer Advisory Panel data collection form including some additional questions. METHODS: The 1-day survey was conducted on March 23, 2004. Each patient was visited by 2 registered nurses, who inspected the patient's skin for any pressure ulcer classified according to the EPUAP grading system. RESULTS: There were no significant differences in gender, age, or Braden score between the patients in surgical, medical, or geriatric care in 2002 and 2004. The overall prevalence of pressure ulcers was 33.3% (grade 1 excluded: 10.9%) in 2002 and 28.2% (grade 1 excluded: 14.1%) in 2004. In surgical care, the prevalence was reduced from 26.8% to 17.3% (P = .051). In medical care, the prevalence was 23.6% in 2002 and 26.7% in 2004. Corresponding prevalence figures for geriatric care were 59.3% and 50.0%. A quarter of the patients in surgical care, a third in medical care, and more than half in geriatric care had a pressure ulcer upon arrival at the ward. The use of pressure-reducing mattresses had increased significantly from 16.0% to 42.7% in medical care (P = .000). CONCLUSIONS: The EPUAP methodology has facilitated the introduction of pressure ulcer as a quality indicator at hospital level. Pressure ulcer prevalence surveys with a standardized methodology should be repeated on a regular basis in order to stimulate quality improvement. PMID- 16717516 TI - Comparison of two pressure ulcer preventive dressings for reducing shear force on the heel. AB - OBJECTIVE: We compared the shear forces exerted over the heel between a pressure ulcer preventive dressing and a thin-film dressing in a clinical setting. Interface pressures were measured as well. DESIGN: Quasi-experimental clinical trial. SETTING AND SUBJECTS: Participants were 30 elderly patients (5 men, 25 women; mean age, 86.4 +/- 8.0 years) hospitalized in a geriatrics hospital in Japan; all had a Braden score of less than 14 (mean, 10.1 +/- 1.1). Informed consent was obtained from all the patients.A shear force and pressure sensor including a strain gauge, and an oval airbag-type pressure sensor were used. METHODS: The sensor was attached to one heel using double-sided tape; then the target dressing was applied over the sensor and on the opposite heel to avoid the influence of the opposite heel on the shear force measurement. Interface pressures were measured with the patient in a stationary supine position; shear force was then measured at 0.2-second intervals, while the sheet was manually pulled at a velocity of around 5 cm/second. Shear force was determined by averaging the stable shear force lasting for 10 measurement points. RESULTS: The mean interface pressures with the PPD and the film dressings were 70.7 +/- 16.5 and 70.2 +/- 15.2 mmHg, respectively; this difference was not statistically significant. The shear force produced during the pulling of the sheet was 2.2 +/- 1.4 and 11.7 +/- 5.8 N, respectively (P < .001, Wilcoxon signed-rank test). CONCLUSIONS: The results of this study suggest that a dressing with a low friction external surface (such as the pressure ulcer preventive dressing) can significantly reduce shear force. However, results also suggest that external dressings do not significantly reduce interface pressures and cannot be used as a substitute for heel elevation in an immobile patient. PMID- 16717517 TI - Pressure ulcer prevention in homecare: do Dutch homecare agencies have an evidence-based pressure ulcer protocol? AB - OBJECTIVE: The purpose of this study was to gain insight into the availability and quality of protocols for pressure ulcer prevention in homecare agencies in the Netherlands. DESIGN: A descriptive study was completed. SETTING AND SUBJECTS: Forty-one homecare agencies in the Netherlands that provide nursing care were queried. INSTRUMENTS: Three instruments were used to collect data: (1) a structured questionnaire containing 46 closed and open-ended questions, (2) a checklist used by experts to analyze the protocols for conformity to guidelines, and (3) a tool used to generate a numerical score for each protocol based on the experts' reviews. METHODS: A questionnaire was mailed to all homecare agencies in the Netherlands that provided nursing care. The quality of each protocol was judged and scored by 3 pressure ulcer prevention experts. The scores were analyzed using descriptive statistics. RESULTS: A pressure ulcer protocol was available in 78% of the agencies. Seventy-five percent had at least 1 wound care nurse who spent an average of 10 hours per week on pressure ulcer prevention. In 20% of the agencies, no introduction or instruction was given to the nurses when the protocol was implemented. In 25% of the agencies, nurses did not participate in the revision of the protocol. At the end of 2003, only 13% of the agencies had executed 1 or more revisions of their protocol since 2002, when the last Dutch pressure ulcer guideline was introduced. The 26 pressure ulcer prevention protocols had a mean score of 47 points out of a maximum of 100 points (range 9 to 82; SD, 18). CONCLUSIONS: Although the use of protocols is considered an important adjunct in the prevention of pressure ulcers, 22% of the participating agencies did not have a pressure ulcer prevention protocol and 25% did not have wound care nurses, indicating a need for further promotion of standardized pressure ulcer prevention strategies. In addition, the available protocols were frequently of low quality or outdated, reflecting a need for increased attention to current and accurate tools to guide nursing practice. PMID- 16717518 TI - Discharge information needs of patients after surgery. AB - Patients who have undergone surgical procedures often have self-care concerns in their preparation for discharge from the hospital. This article examines the research literature about information needs of postoperative patients prior to their discharge. The most common concerns were the incision/wound care, pain management, activity level, monitoring for complications, symptom management, elimination, and quality of life. Because of their clinical knowledge of the perioperative experience, wound, ostomy, and continence nurses and other advanced practice nurses have a critical role in the development of discharge-educational programs for postoperative patients and caregivers. Because unmet discharge needs can contribute to poor patient outcomes and readmission, it is critical that wound, ostomy, and continence nurses, advanced practice nurses, and clinical staff nurses accurately identify patients' informational needs and find ways to meet these needs especially with the aging population, new/advanced surgical procedures, vulnerability/poverty, and literacy level of patients. PMID- 16717520 TI - Are true-negative nitrite results affected by a two-hour delay in dipstick/pad analysis of urine in incontinence pads? AB - OBJECTIVES: To determine if true-negative nitrite results of a urine dipstick pressed into an incontinence pad (dipstick/pad method) are affected by a 2-hour delay in analysis. DESIGN: A quantitative study. SETTING AND SUBJECTS: Clean catch urine specimens from a convenience sample of clinic patients, staff, and long-term care facility residents. INSTRUMENT: Changes from negative to positive for each group of urine specimens were evaluated using percentages within the groups. METHODS: Urine specimens were collected and a portion was cultured. Urine specimens negative for a urinary tract infection were included in the study. A portion of the specimen was poured into an incontinence pad. Initial nitrite results were determined using a dipstick pressed into an incontinence pad. Pads with true-negative dipstick/pad nitrite results were tested 2 hours later in the same manner. Urine culture results determined groups: mixed colonies; 50,000 to 75,000 colony-forming units per milliliter of a single type of uropathogen; greater than or equal to 50,000 cfu/mL diphtheroids; no significant growth; and no growth. RESULTS: Of the 443 urine cultures negative for a urinary tract infection, 441 initial dipstick/pad nitrite results were negative. Two initial true-negative nitrite dipstick/pad results, or 0.5%, changed from negative to positive over the 2-hour period: 1 in the "mixed colonies" group (0.4%) and 1 in the "no growth" group (0.7%). CONCLUSIONS: Results of this study indicate that true-negative nitrite results of a dipstick pressed into urine in an incontinence pad do not appear to be affected by a 2-hour delay in analysis. PMID- 16717521 TI - Tag F315: an opportunity for WOC nurses. AB - Urinary incontinence affects more than 50% of elderly patients in hospitals and in nursing homes. Of the estimated $16.3 billion spent annually, 90% is spent on management, whereas only 10% is spent on diagnosis and treatment. With their June 2005 release of the F315 tag, the Centers for Medicare and Medicaid Services are taking steps to change the circumstances of this disorder. This tag provides several guidelines for continence care. One of the most noticeable mandates is that of "restoring as much normal bladder function as possible." This mandate provides various novel opportunities for the WOC nurses and advanced practice nurses with WOC certification. PMID- 16718266 TI - Melanoma, Parkinson's disease and levodopa: causal or spurious link? A review of the literature. AB - Since the early 1970s, a number of case reports have suggested that levodopa therapy for Parkinson's disease increases the risk of cutaneous malignant melanoma. As yet, no formal epidemiological study has been conducted to verify this hypothesis. To elucidate the relationship between levodopa and the risk of cutaneous malignant melanoma, a systematic literature search using computerized bibliographic databases was done. This review presents the case history evidence for and against the hypothesis of a causal association, and explores possible epidemiological, genetic, social, biochemical and toxicological factors that may increase the risk of melanoma in Parkinson's disease patients. All the case reports in the literature were considered. We concluded that (1) there is no epidemiological or experimental evidence of a causal role of levodopa in increasing the risk of melanoma incidence or progression; (2) there is good evidence of an excess risk of melanoma in patients with Parkinson's disease; (3) there is good evidence of a protective effect of tobacco smoking on the risk for Parkinson's disease; (4) there is good evidence of positive correlation between social class and melanoma risk; (5) the relationship between the risk of Parkinson's disease and the risk of melanoma may be due to a common genetic profile or it can be attributed to a confounding role of social class, associated with both melanoma and Parkinson's disease possibly through an inverse relationship with tobacco smoking. PMID- 16718267 TI - Induction of secreted matrix metalloproteinase-9 activity in human melanoma cells by extracellular matrix proteins and cytokines. AB - Degradation of matrix proteins that constitute the dermal-epidermal junction and dermis by proteolytic enzymes is an essential step of melanoma invasion and metastasis, and this is primarily achieved by the matrix metalloproteinases. In this report, using zymography, we compared the basal secretion levels of active matrix metalloproteinase-2 and matrix metalloproteinase-9 to levels in response to various extracellular matrix proteins, cytokines, and growth factors in normal human melanocyte cells and melanoma cell lines from different stages of neoplastic progression. Basal matrix metalloproteinase-9 activity was only detected in vertical growth phase and metastatic melanoma cell lines, suggesting that matrix metalloproteinase-9 is a candidate biomarker for identifying vertical growth phase and metastatic melanomas. Most melanoma cell lines and cultured normal melanocytes produced high levels of matrix metalloproteinase-2. In addition, both tumor necrosis factor-alpha and interleukin-1beta are strong inducers of active matrix metalloproteinase-9 in vertical growth phase melanoma cell lines, indicating a possible role of these cytokines in the switch from radial growth phase to vertical growth phase. We propose that these proinflammatory cytokines promote melanoma invasion in part through upregulating matrix metalloproteinase-9. Both these cytokines are released from keratinocytes in the epidermis by ultraviolet radiation. Thus, our study suggests that the microenvironment of melanoma cells is an important feature in melanoma progression, and ultraviolet-radiation-induced cytokines might promote the progression of melanoma through the release or activation of matrix metalloproteinases. PMID- 16718268 TI - Expression of ecto-5'-nucleotidase (eN, CD73) in cell lines from various stages of human melanoma. AB - Ecto-5'-nucleotidase is a GPI-anchored enzyme localized in cell membrane lipid rafts. Although it is highly expressed in many tumour cells, its specific function during tumorigenesis is unclear. We have found that, among different melanoma cells, upregulated expression of ecto-5'-nucleotidase is associated with a highly invasive phenotype. Analysis of other cell membrane proteins involved in melanoma adhesion and metastasis demonstrated that expression of alpha5, beta1, beta3-integrin subunits and CD44 was elevated gradually in accordance with increasing metastatic potential. Expression of alphav-integrin and caveolin-1 was seen mostly in cells derived from metastatic melanomas. Furthermore, in contrast to N-cadherin, which was unaltered in all lines, we could not detect E-cadherin in any cell type. Functional assays demonstrated that highly expressed ecto-5' nucleotidase is a catalytically competent protein that is very sensitive to inhibition by concanavalin A. The interaction with concanavalin A also caused increased association of ecto-5'-nucleotidase-rich lipid rafts with much heavier cytoskeletal complexes as determined by density gradient centrifugation. A similar shift towards heavier cytoskeletal fractions also took place with other proteins coexpressed with ecto-5'-nucleotidase, such as alphav, alpha5, beta1 and beta3-integrins, caveolin-1 and CD44. As ConA-induced clustering may reflect the interactions of membrane proteins with extracellular matrix, we also analysed the effect of several extracellular matrix proteins on the in-situ activity of ecto 5'-nucleotidase in WM9 cells and found that tenascin C strongly inhibited ecto-5' nucleotidase activity and adenosine generation from AMP. We also developed WM9 cells with reduced ecto-5'-nucleotidase expression and tested differences in cell adhesion on various extracellular matrix proteins. WM9 cells attached significantly weaker to tenascin C layer. These observations indicate that expression of ecto-5'-nucleotidase correlates with a number of metastasis-related markers and thus may have a function in this process. Furthermore, our data suggest that, in addition to generating adenosine, ecto-5'-nucleotidase may have independent roles in adhesion and interaction with extracellular matrix components in melanoma. PMID- 16718269 TI - Expression of the human endogenous retrovirus-K transmembrane envelope, Rec and Np9 proteins in melanomas and melanoma cell lines. AB - The human endogenous retrovirus-K encodes two potential tumor proteins, Rec and Np9. Rec is related to the Rev protein of HIV-1 and has been shown to be associated with tumor development in nude mice. Having shown the expression of human endogenous retrovirus-K in human melanomas and melanoma cell lines, tools were developed to allow the expression of the transmembrane envelope, Rec and Np9 mRNA and proteins to be studied in more detail. The expression of spliced env, rec and np9 was investigated by reverse transcriptase-polymerase chain reaction using a set of primers developed to discriminate between full-length and spliced mRNA. Env-specific, Rec-specific and Np9-specific antisera were produced, characterized and used to study protein expression in melanomas and melanoma cell lines by immunohistochemistry, immunofluorescence and Western blot analyses. Existence of human endogenous retrovirus-K Rec and Np9-specific antibodies in the sera of melanoma patients were analyzed by Western blot of immunofluorescence studies. The expression of both spliced env and rec mRNA was detected in 39% of the melanomas and in 40% of the melanoma cell lines and np9 mRNA was detected in 29 and 21%, respectively. In normal neonatal melanocytes, spliced rec mRNA was detected in the absence of spliced env mRNA. Using antisera specific for Rec and Np9, Rec protein was found in 14% of the melanomas but Np9 in none. In addition, cell surface expression of the putatively immunosuppressive transmembrane envelope protein and release of virus particles were shown. Antibodies specific for neither Rec nor Np9 were detected. The transmembrane envelope protein, Rec and Np9 proteins are expressed in melanoma cells with a pattern similar to that seen in teratocarcinoma cell lines. Additional experiments are needed to determine their involvement, if any, in cell proliferation and tumor progression. PMID- 16718270 TI - Different expression of lysosome-associated membrane protein-1 in human melanomas and benign melanocytic lesions. AB - Lysosome-associated membrane protein-1 is a protein with a significant content of beta1,6-branched N-glycans. It is thought that enhanced expression of lysosome associated membrane protein-1 in tumour cells may promote invasion by influencing both adhesion to extracellular matrix and perhaps also binding to endothelial cells. The present study was aimed at examining levels of lysosome-associated membrane protein-1 in human melanomas and benign pigmented lesions to evaluate whether this protein might be considered a potential molecular marker of melanoma progression. The expression of lysosome-associated membrane protein-1 was for the first time determined immunohistochemically in formalin-fixed paraffin-embedded specimens comprising 42 primary cutaneous melanomas, 15 lymph node melanoma metastases (11 correlated with primary tumours), three melanoma recurrences (correlated with both primary and metastatic melanomas), 27 nevi and four epithelial tumours (two seborrhoeic keratoses and two basal cell carcinomas). Our results demonstrate that development and progression of melanoma are associated with changes of the lysosome-associated membrane protein-1 level. The expression was strongest in melanoma recurrences and lymph node metastases, weaker in primary cutaneous melanomas and not detectable in melanocytes of pigmented nevi. Nodular melanomas expressed lysosome-associated membrane protein-1 at higher level than superficially spreading melanomas. PMID- 16718271 TI - A prospective single arm phase II study of dacarbazine and treosulfan as first line therapy in metastatic uveal melanoma. AB - Uveal melanoma is relatively uncommon accounting for fewer than 5% of all melanoma cases. Localized tumours are curable by local therapy but a significant percentage of patients go on to have a relapse with metastatic disease. Uncertainty remains concerning the level of activity of dacarbazine in uveal melanoma as opposed to that in the cutaneous form. Recently, a possible role for treosulfan in uveal disease has been reported. A phase II study was therefore undertaken to assess the objective response rate of the combination of dacarbazine and treosulfan in previously untreated patients with metastatic uveal melanoma. All patients received dacarbazine 850 mg/m and treosulfan 8 g/m(2) every 21 days up to a maximum of six cycles. Fifteen patients enrolled in the study. As expected, the major toxicities were haematological (particularly thrombocytopaenia) but the treatment was generally well tolerated. No responses were seen; however, disease stabilization was achieved in two patients. Median progression free survival from the start of chemotherapy was 12 weeks and median overall survival was 30 weeks. This study, using the combination of dacarbazine and treosulfan, while well tolerated, did not confirm earlier reports suggesting treosulfan is active in uveal melanoma. PMID- 16718272 TI - Possibilities of improving the parameters of hyperthermia in regional isolated limb perfusion using epidural bupivacaine and accurate temperature measurement of the three layers of limb tissue. AB - The present study presents the author's modification of the method, which aims to create proper parameters of the treatment. The selected group consisted of 15 women and eight men, with a mean age of 57.2 years (range from 26 to 72 years). The patients were divided into two groups, depending on whether they were given epidural bupivacaine (group I - 13 patients treated between the years 2001 and 2004) or not [group II (control) - 10 patients treated earlier, between the years 1997 and 2000]. We observed a significant change in the temperature of thigh muscles (P=0.009) and shank muscles (P=0.006). In the control group II, there was a statistically significant difference (P=0.048) in the temperatures between the muscles and subcutaneous tissue on the one hand and the shank skin on the other. That difference was mean 0.67 degrees Celsius (from 0.4 to 0.9) during the perfusion after applying the cytostatic. The temperature of the skin was lower than the temperature of the deeper tissues of the shank and did not exceed 39.9 degrees Celsius. Such a difference in the temperatures was not observed in case of the group I patients who were given bupivacaine into the extrameningeal space before applying the cytostatic. The difference in the temperatures was on average 0.26 degrees Celsius and was not statistically significant (P=0.99), whereas the shank skin temperature was 40.0-40.6 degrees Celsius. The attained results imply that despite the noticeable improvement in the heating of the limb muscles after application of bupivacaine, the improvement in the heating of the skin and subcutaneous tissue is still not satisfactory, although the growing tendency implies such a possibility. PMID- 16718273 TI - Penile metastasis of cutaneous malignant melanoma: a true hematogenous spread?: Case report and review of the literature. AB - Penile involvement has been implicated as a metastatic site in several tumors; approximately 300 cases have been reported. Of these, only two cases showed cutaneous melanoma as the primary site. Our patient presented with a painless mass on the penile shaft together with other distant metastases. A magnetic resonance image demonstrated two sites of deposit in the subcutaneous tissue on the radix penis, and fine needle aspiration cytology of the mass confirmed the presence of melanoma cells. The patient died of systemic disease without any further treatment for penile involvement. This unusual involvement is presented with a review of the related literature. PMID- 16718274 TI - Complete and long-lasting regression of disseminated multiple skin melanoma metastases under treatment with cyclooxygenase-2 inhibitor. AB - Experimental and clinical evidence indicates that non-steroidal anti-inflammatory drugs and cyclooxygenase-2 inhibitors may have anti-cancer activities. Here we report on a patient with a metastatic melanoma of the leg who experienced a complete and sustained regression of skin metastases upon continuous single treatment with the cyclooxygenase-2 inhibitor rofecoxib. Our observations indicate that the inhibition of cyclooxygenase-2 can lead to the regression of disseminated skin melanoma metastases, even after failure of chemotherapy. PMID- 16718275 TI - Diagnostics and biomarker development: priming the pipeline. AB - The decrease in the rate at which novel medical products are reaching the market, despite major scientific achievements and investment that might have predicted otherwise, is causing much concern. Although this 'pipeline problem' has often been discussed in the context of drug development, it is also crucial to examine the unique characteristics of the pipeline for biomarkers and diagnostics. Here, we characterize the pipeline problem for biomarkers and diagnostics, and consider what steps could be taken to solve it. PMID- 16718276 TI - Electrical stimulation-evoked resistance exercise therapy improves arterial health after chronic spinal cord injury. AB - STUDY DESIGN: Repeated measures training intervention. OBJECTIVES: To evaluate the effects of neuromuscular electrical stimulation (NMES)-induced resistance exercise therapy on lower extremity arterial health in individuals with chronic, complete spinal cord injury (SCI). We define "arterial health" using three surrogate markers: (a) resting diameter, (b) flow-mediated dilation (FMD), and (c) arterial range. SETTING: Department of Kinesiology, University of Georgia, USA. METHODS: We assessed five 36+/-5-year-old male individuals with chronic, complete SCI before, during, and after 18 weeks of training. The quadriceps femoris muscle group of both legs were trained twice a week with 4 x 10 repetitions of unilateral, dynamic knee extensions. The health of the posterior tibial artery was assessed using a B-mode ultrasound unit equipped with a high resolution video capture device. Proximal occlusion was used to evoke ischemia for 5 min and then for 10 min. FMD was calculated using the peak diameter change (above rest) following 5 min occlusion. Arterial range was calculated using minimum (during occlusion) and maximum diameters (post 10 min occlusion). Hierarchical linear modeling accounted for the nested (repeated measures) experimental design. RESULTS: FMD improved from 0.08+/-0.11 mm (2.7%) to 0.18+/ 0.15 mm (6.6%) (P=0.004), and arterial range improved from 0.36+/-0.28 to 0.94+/ 0.40 mm (P=0.001), after 18 weeks of training. Resting diameter did not significantly change. CONCLUSIONS: Home-based, self-administered NMES resistance exercise therapy consisting of 80 contractions/week improved FMD and arterial range. This provides evidence that resistance exercise therapy can improve arterial health after SCI, which may reduce the risk of future cardiovascular disease. PMID- 16718277 TI - Effects of antibacterial therapy on PSA change in the presence and absence of prostatic inflammation in patients with PSA levels between 4 and 10 ng/ml. AB - We investigated the effect of prostatic inflammation on prostate-specific antigen (PSA) and per cent-free PSA levels changing after antibacterial therapy. We evaluated 48 patients whose PSA levels were between 4 and 10 ng/ml, without any suspicious findings on digital rectal examination, with no infection findings in urine analysis. Prostatic inflammation was assessed with prostatic massage. All the patients were given antibiotic therapy for 3 weeks. Patients were re evaluated 3 weeks after antibacterial therapy with PSA (free/total) and urinalysis. Ten core biopsies were taken with transrectal ultrasound. No differences were found in terms of age, pre- and post-treatment PSA, and PSA varying between patients with and without inflammation in the prostatic massage. In 18 patients, PSA decreased below 4 ng/ml. Prostate cancer was found in 10.8% of the patients with PSA between 4 and 10 ng/ml and none of the patients with PSA values below 4 ng/ml. We suggest an antibiotic therapy for 3 weeks without regarding inflammation findings when PSA is in the gray zone, for biopsy decision. PMID- 16718279 TI - Ontogenic reduction of Aph-1b mRNA and gamma-secretase activity in rats with a complex neurodevelopmental phenotype. AB - Selectively bred apomorphine susceptible (APO-SUS) rats display a complex behavioral phenotype remarkably similar to that of human neurodevelopmental disorders, such as schizophrenia. We recently found that the APO-SUS rats have only one or two Aph-1b gene copies (I/I and II/II rats, respectively), whereas their phenotypic counterpart has three copies (III/III). Aph-1b is a component of the gamma-secretase enzyme complex that is involved in multiple (neuro)developmental signaling pathways. Nevertheless, surprisingly little is known about gamma-secretase expression during development. Here, we performed a longitudinal quantitative PCR study in embryos and the hippocampus of I/I, II/II and III/III rats, and found gene-dosage dependent differences in Aph-1b, but not Aph-1a, mRNA expression throughout pre- and post-natal development. On the basis of the developmental mRNA profiles, we assigned relative activities to the various Aph-1a and -1b gene promoters. Furthermore, in the three rat lines, we observed both tissue-specific and temporal alterations in gamma-secretase cleavage activity towards one of its best-known substrates, the amyloid-beta precursor protein APP. We conclude that the low levels of Aph-1b mRNA and gamma secretase activity observed in the I/I and II/II rats during the entire developmental period may well underlie their complex phenotype. PMID- 16718280 TI - Association between glutamic acid decarboxylase genes and anxiety disorders, major depression, and neuroticism. AB - Abnormalities in the gamma-aminobutyric acid (GABA) neurotransmitter system have been noted in subjects with mood and anxiety disorders. Glutamic acid decarboxylase (GAD) enzymes synthesize GABA from glutamate, and, thus, are reasonable candidate susceptibility genes for these conditions. In this study, we examined the GAD1 and GAD2 genes for their association with genetic risk across a range of internalizing disorders. We used multivariate structural equation modeling to identify common genetic risk factors for major depression, generalized anxiety disorder, panic disorder, agoraphobia, social phobia and neuroticism (N) in a sample of 9270 adult subjects from the population-based Virginia Adult Twin Study of Psychiatric and Substance Use Disorders. One member from each twin pair for whom DNA was available was selected as a case or control based on scoring at the extremes of the genetic factor extracted from the analysis. The resulting sample of 589 cases and 539 controls was entered into a two-stage association study in which candidate loci were screened in stage 1, the positive results of which were tested for replication in stage 2. Several of the six single-nucleotide polymorphisms tested in the GAD1 region demonstrated significant association in both stages, and a combined analysis in all 1128 subjects indicated that they formed a common high-risk haplotype that was significantly over-represented in cases (P=0.003) with effect size OR=1.23. Out of 14 GAD2 markers screened in stage 1, only one met the threshold criteria for follow-up in stage 2. This marker, plus three others that formed significant haplotype combinations in stage 1, did not replicate their association with the phenotype in stage 2. Subject to confirmation in an independent sample, our study suggests that variations in the GAD1 gene may contribute to individual differences in N and impact susceptibility across a range of anxiety disorders and major depression. PMID- 16718281 TI - Perinatal and childhood origins of cardiovascular disease. AB - BACKGROUND: Features of the metabolic syndrome comprise a major risk for cardiovascular disease and will increase in prevalence with rising childhood obesity. We sought to identify early life influences on development of obesity, hypertension and dyslipidemia in children. METHODS AND RESULTS: Cluster analysis was used on a subset of a longitudinal Australian birth cohort who had blood samples at age 8 (n=406). A quarter of these 8-year-olds fell into a cluster with higher body mass index, blood pressure (BP), more adverse lipid profile and a trend to higher serum glucose resembling adult metabolic syndrome. There was a U shaped relationship between percentage of expected birth weight (PEBW) and likelihood of being in the high-risk cluster. The high-risk cluster had elevated BP and weight as early as 1 and 3 years old. Increased likelihood of the high risk cluster group occurred with greatest weight gain from 1 to 8 years old (odds ratio (OR)=1.4, 95% confidence interval (CI)=1.3-1.5/kg) and if mothers smoked during pregnancy (OR=1.82, CI=1.05-3.2). Risk was lower if children were breast fed for >/=4 months (OR=0.6, 95% CI=0.37-0.97). Newborns in the upper two quintiles for PEBW born to mothers who smoked throughout pregnancy were at greatest risk (OR=14.0, 95% CI=3.8-51.1) compared to the nadir PEBW quintile of non-smokers. CONCLUSION: A U-shaped relationship between birth weight and several components of the metabolic syndrome was confirmed in a contemporary, well nourished Western population of full-term newborns, but post-natal weight gain was the dominant factor associated with the high-risk cluster. There was a prominence of higher as well as lowest birth weights in those at risk. Future health programs should focus on both pre- and post-natal factors (reducing excess childhood weight gain and smoking during pregnancy), and possibly the greatest benefits may arise from targeting the heaviest, as well as lightest newborns, especially with a history of maternal smoking during pregnancy. PMID- 16718282 TI - The effect of a medium-term activity- and diet-induced energy deficit on subjective appetite sensations in obese children. AB - OBJECTIVE: To assess the subjective appetite responses to an imposed activity- and diet-induced energy deficit during a residential intervention programme for obese children. DESIGN: A 6-week intervention of fixed, reduced dietary intake and 6 h/day of skill-based physical activity while resident in a weight loss camp. SUBJECTS: Thirty-eight obese (mean body mass index (BMI)=34.9 kg m(-2)) boys and girls (mean age 13.9+/-1.57). MEASUREMENTS: An electronic appetite rating system (EARS) was used to periodically measure subjective appetite sensations at the start (WK1) and at the end (WK6) of the camp. Subjective ratings of hunger and fullness were compared at the start and end of 6 weeks of an activity- and diet-induced-based weight loss intervention. RESULTS: At the end of the 6 weeks, the children had lost 8.4 kg in body mass. The diurnal profiles of subjective appetite sensations demonstrated clear oscillations in hunger and fullness. There was a significant increase in hunger (P<0.0001) and decrease in fullness (P<0.005) at the end (WK6) of the medium-term energy deficit. In WK6, morning ratings of hunger were higher than in WK1 (P<0.005) and the fixed energy evening meal induced a lower suppression of hunger (P<0.02). CONCLUSION: A programme of fixed, reduced-dietary intake combined with an activity and behavioural programme was successful at inducing a significant reduction in body mass in obese children. Subjective sensations of appetite were sensitive to a medium-term negative energy balance and weight loss. These data are essential as we continue to evolve methods of treatment for overweight and obese children. PMID- 16718283 TI - Association between sleeping hours, working hours and obesity in Hong Kong Chinese: the 'better health for better Hong Kong' health promotion campaign. AB - OBJECTIVE: To study the inter-relationships between sleeping hours, working hours and obesity in subjects from a working population. RESEARCH DESIGN: A cross sectional observation study under the 'Better Health for Better Hong Kong' Campaign, which is a territory-wide health awareness and promotion program. SUBJECTS: 4793 subjects (2353 (49.1%) men and 2440 (50.9%) women). Their mean age (+/-s.d.) was 42.4+/-8.9 years (range 17-83 years, median 43.0 years). Subjects were randomly selected using computer-generated codes in accordance to the distribution of occupational groups in Hong Kong. RESULTS: The mean daily sleeping time was 7.06+/-1.03 h (women vs men: 7.14+/-1.08 h vs 6.98+/-0.96 h, P<0.001). Increasing body mass index (BMI) was associated with reducing number of sleeping hours and increasing number of working hours reaching significance in the whole group as well as among male subjects. Those with short sleeping hour (6 h or less) and long working hours (>9 h) had the highest BMI and waist in both men and women. Based on multiple regression analysis with age, smoking, alcohol drinking, systolic and diastolic blood pressure, mean daily sleeping hours and working hours as independent variables, BMI was independently associated with age, systolic and diastolic blood pressure in women, whereas waist was associated with age, smoking and blood pressure. In men, blood pressure, sleeping hours and working hours were independently associated with BMI, whereas waist was independently associated with age, smoking, blood pressure, sleeping hours and working hours in men. CONCLUSION: Obesity is associated with reduced sleeping hours and long working hours in men among Hong Kong Chinese working population. Further studies are needed to investigate the underlying mechanisms of this relationship and its potential implication on prevention and management of obesity. PMID- 16718284 TI - Cultural conflicts in the weight loss experience of overweight Latinos. AB - OBJECTIVE: In spite of the high prevalence of obesity in the Latino population, there is limited recent information that can be used by health-care providers to develop culturally appropriate weight loss strategies for this population. Therefore, we describe weight loss experiences, attitudes and barriers in overweight Latino adults. DESIGN: Qualitative study using focus group methodology. SUBJECTS: Twenty-one overweight adults (body mass index >/=25, age >/=20 years) self-identified as Latinos. METHODS: Subjects participated in one of three focus groups. Reccurring themes within group discussions were identified by three independent investigators, one who was ethnicity concordant. RESULTS: Themes included the presence of mixed messages when determining one's appropriate weight, with participants' desire to lose weight to be healthy (based on professional advice and personal experience) conflicting with the cultural idea that being overweight is healthy. Participants described discordance when adapting to the mainstream, leading to the loss of healthy traditional habits. Participants expressed interest in weight loss and familiarity with dieting and weight loss interventions. They desired culturally appropriate nutrition education and reassurance regarding healthy dieting from health-care providers. The importance of interactions with peers during education was another relevant theme, and participants were overwhelmingly positive about group education. CONCLUSIONS: To improve health promotion for Latinos, cultural factors distinctive to this underserved population, and barriers they articulate, should be considered when developing weight loss interventions. PMID- 16718286 TI - Body mass, weight control behaviours, weight perception and emotional well being in a multiethnic sample of early adolescents. AB - OBJECTIVE: To investigate weight perception, dieting and emotional well being across the range of body mass index (BMI) in a population-based multiethnic sample of early adolescents. DESIGN: Cross-sectional population-based survey. SUBJECTS: In total, 2789 adolescents 11-14 years of age from three highly deprived regional authorities in East London, in 2001. MEASUREMENTS: Data were collected by student-completed questionnaire on weight perception, dieting history, mental and physical health, health behaviours, social capital and sociodemographic factors. Height and weight were measured by trained researchers. Overweight was defined as BMI > or =85th centile and obesity as BMI > or =98th centile. Underweight was defined as BMI< or =15th centile. RESULTS: In all, 73% were from ethnic groups other than white British. Valid BMI were available for 2522 subjects (90.4%) of whom 14% were obese. Only 20% of overweight boys and 51% of overweight girls assessed their weight accurately. Accuracy of weight perception did not vary between ethnic groups. In all, 42% of girls and 26% of boys reported current dieting to lose weight. Compared with white British teenagers, a history of dieting was more common among Bangladeshi, Indian and mixed ethnicity boys and less likely among Pakistani girls. Self-esteem was not associated with BMI in girls but was significantly lower in obese boys than those of normal weight (P=0.02). Within ethnic subgroups, self-esteem was significantly lower in overweight white British boys (P=0.03) and obese Bangladeshi boys (P=0.01) and Bangladeshi girls (P=0.04), but significantly higher in obese black African girls (P=0.01) than those of normal weight. Obese young people had a higher prevalence of psychological distress (P=0.04), except among Bangladeshi teenagers, where overweight and obese young people had less psychological distress than those of normal weight (P=0.02). Birth outside the UK was associated with reduced risk of obesity in girls (P=0.02) but not with history of dieting, weight perception or psychological factors in either gender. CONCLUSION: High levels of current dieting for weight control and inaccurate perception of body mass are common across all ethnic groups. However, dieting history and the associations of obesity with self-esteem and psychological distress vary between ethnic groups. Interventions to prevent or treat obesity in black or minority ethnicity groups must consider cultural differences in the relationship between body mass, self-esteem and psychological distress. PMID- 16718285 TI - The association between early childhood stunting and weight status in late adolescence. AB - BACKGROUND: Cross-sectional studies have shown associations between stunting and overweight; however, there are few prospective studies of stunted children. OBJECTIVES: To determine whether stunting before age 2 years is associated with overweight and central adiposity at 17-18 years and whether growth in height among stunted children predicts body mass index (BMI) in late adolescence. DESIGN: Prospective cohort study. PARTICIPANTS: One-hundred and three participants stunted by age 2 years and 64 non-stunted participants (78% of participants enrolled in childhood). Participants were measured in early childhood and at ages 7, 11 and 17 years. RESULTS: Stunted subjects remained shorter and had lower BMIs, smaller skinfolds and circumferences than non-stunted subjects. Overweight (BMI >/=25 m(2)) was not significantly different among stunted and non-stunted male subjects (5.2 and 12.5%) but non-stunted female subjects were more likely to be overweight than those who experienced early childhood stunting (11.1 and 34.4%, P=0.013). Centralization of fat (waist to hip ratio (WHR), subscapular/triceps skinfold ratio (SSF/TSF)) did not differ between stunted and non-stunted groups (mean WHR 0.77 and mean SSF/TSF 1.18 in both groups). Stunted subjects with greater increases in height-for-age for the intervals 3-7 and 7-11 years had higher BMI at age 17 years (P=0.04 and P=0.001, respectively). CONCLUSION: Participants stunted by age 2 years were less likely to be overweight than those who were never stunted. This suggests that cross sectional studies of the association between stunting and overweight may be misleading. Among stunted children, greater linear growth during mid- to late childhood was associated with greater BMI at age 17 years. PMID- 16718287 TI - Nocturia in adults: draft New Zealand guidelines for its assessment and management in primary care. AB - Nocturia is a common bothersome condition. An ad hoc group of interested clinicians from a variety of backgrounds has developed draft guidelines for the assessment and management of this condition in primary care in New Zealand. The guidelines propose four steps in the assessment and management: clinical evaluation; simple investigations; assignment of a provisional diagnosis; and management based on the provisional diagnosis. For nocturnal polyuria-associated nocturia, the draft guidelines recommend that: lifestyle measures should be used as part of the management; if a patient complaining of nocturia has other features of overactive bladder, then bladder retraining and/or anticholinergics can be used; hypnosedatives should not be used to treat nocturia in older adults because of the increased risk of falls; loop diuretics given in the afternoon should be considered for the treatment; and desmopressin can be considered in the management of nocturnal polyuria associated nocturia but that it should be used cautiously in people aged over 65 because of the risk of hyponatraemia. A draft algorithm based on international guidelines is presented. PMID- 16718288 TI - Current and former smoking increases mortality in patients on peritoneal dialysis. AB - AIMS: There is limited information on the effects of smoking behaviour on mortality in patients with end-stage renal failure (ESRF). This study aimed to assess the interaction of smoking on death rate in patients with renal failure on dialysis. METHODS: All patients (n=1293) commencing peritoneal dialysis between 1985 and 1995 for renal failure in New Zealand were prospectively followed 6 monthly until 1997 and data entered on the National database. Mortality rates were calculated from the national database and rates in patients with diabetes compared with those without diabetes and in those who did or did not smoke. RESULTS: Follow-up data was available on all patients for a range of 20-40 months. 35% of the patients were clinically classified as having diabetic nephropathy as the cause of renal failure (11% type 1, 24% type 2). Seventeen percent of the total cohort were current smokers, 45% former smokers and 38% lifetime non smokers at dialysis commencement. These rates were similar between patients with diabetes (18% current, 51% former, 32% non-smoker) and those without diabetes (17% current, 42% former, 41% non-smoker). At survey end in 1997, 43% of the patients without diabetes had died compared with 59% of patients with type 1 diabetes (p<0.05) and 62% of patients with type 2 diabetes (p<0.05). The age-adjusted mortality of patients with a history of current or former smoking was higher than non-smokers. Those patients with diabetes and a history of smoking had even higher mortality. CONCLUSIONS: Patients with a current or former history of smoking on peritoneal dialysis are at greatly increased risk of death. A strategy of aggressive smoking cessation efforts should be adopted for these patients at the earliest opportunity. PMID- 16718289 TI - Maori have a much higher incidence of community-acquired pneumonia and pneumococcal pneumonia than non-Maori: findings from two New Zealand hospitals. AB - To determine the incidence rates of community-acquired pneumonia and pneumococcal pneumonia requiring hospitalisation among Maori and non-Maori, an observational study was conducted in Christchurch and Hamilton. Self-reported data were collected using an interviewer-administered questionnaire. Routine clinical, radiological, and microbiological techniques were used apart from the BinaxNow pneumococcal antigen test for diagnosis of this infection. Census data was used to determine the denominator for statistical analyses. The pneumonia rate overall was 3.03 times higher among Maori than non-Maori (p<0.001). Differences were significant for each 10-year age group from age 45-74 years (p<0.05). The rate of pneumococcal pneumonia was 3.23 fold higher for Maori than non-Maori (p<0.001), but it did not reach statistical significance in the age-related comparisons. These ethnic disparities are of major concern, and policy planners should consider further interventions to improve the efficacy of current anti-smoking campaigns and to undertake studies of conjugate pneumococcal vaccines for Maori. PMID- 16718290 TI - After hours healthcare for older patients in New Zealand. AB - AIMS: To explore patterns of general practice after hours service use in different age groups, and to identify possible reasons for any differences between older and younger people in their use of after hours services. METHODS: Attendance data from two after hours clinics (AHCs) and the local hospital emergency department (ED) for 2002 were collected and analysed statistically. RESULTS: Older people, especially the "young-old" (aged 65 to 74) used after hours health centres at a lower rate than younger adults. Older people attended the ED at a higher rate than younger adults, except for the "young-old" who attended at a similar rate. European ethnicity, symptoms resulting from an accident, and increasing severity of the illness were positively correlated with ED attendance. CONCLUSIONS: Older people presented to after hours clinics less than would be expected, especially given their greater morbidity. Older people attended the Hutt Valley ED after hours at a higher rate than younger people, but at a lower rate than in normal hours. Older people were sicker on arrival at ED (especially after hours) than younger people. These results have implications for service delivery and also equity issues as they affect older people. PMID- 16718291 TI - Older people and after hours care. PMID- 16718292 TI - Hip fractures: a deadly and silent epidemic. PMID- 16718293 TI - Neurophysiological findings in a case of cervical anterior spinal artery syndrome: compound muscle action potentials, a marker for prognosis. AB - We report a case of cervical anterior spinal artery syndrome (ASAS). MRI showed abnormal hypointense on T1-weighted images and hyperintense on T2-weighted images from vertebrae C2 to T3. The lesion involved the anterior two-third bilaterally. Spinal angiography showed the superior segmental obstruction of the anterior spinal artery. Regarding nerve conduction studies, no CMAP (the compound muscle action potentials) could be obtained in either median nerves or ulnar nerves, and F-waves were absent. Six months after the onset, there was no any recovery of strength in both arms of the ASAS patient. In our opinion, CMAP could be seen a marker of prognosis for ASAS patients, and absent CMAP might forecast the bad prognosis. PMID- 16718294 TI - Surgical inclination in senior medical students from the University of Auckland: results of the 2005 Senior Students Survey. AB - AIMS: To determine the proportion of senior medical students who are surgically inclined and to assess whether gender differences exist in surgical inclination. STUDY DESIGN: Cross-sectional survey. Twenty-five point questionnaire. Likert scale response ranking. SETTING: University of Auckland Medical School, New Zealand. PARTICIPANTS: 218 surveys were emailed to functioning addresses of fourth and fifth year students.156 students emailed responses (71.60% response rate). RESULTS: Twenty percent of students were found to be surgically inclined (95% CI 0.15-0.26). The proportion of surgically inclined males was significantly higher than females (p<0.01). A greater proportion of surgically inclined students found time spent in the operating theatre educationally valuable than non-surgically inclined students (p<0.01). No difference exists in the number of different procedures undertaken by students (p<0.05). CONCLUSION: Males are significantly more likely to be surgically inclined than females at the University of Auckland Medical School. PMID- 16718295 TI - Portal venous thrombophlebitis in a case of perforated appendicitis: lessons from a case. AB - Portal and mesenteric pyelephlebitis is a rarely recognised condition associated with a high morbidity. It usually develops secondary to infection in the drainage area of the portal venous system. We report a case of perforated acute appendicitis complicated by superior mesenteric venous pyelephlebitis and thrombocytopaenia. Appendicectomy and treatment with broad-spectrum antibiotics, anticoagulation, and platelets led to a full recovery. Follow-up imaging revealed complete resolution of the thrombosis. The literature is reviewed and the operative and non-operative approaches for the management of mesenteric and portal venous thrombosis are discussed. PMID- 16718296 TI - Medical image. Colon obstruction. PMID- 16718298 TI - In defence of Ayurvedic medicine. PMID- 16718299 TI - Buteyko breathing technique and asthma in children: a case series. PMID- 16718297 TI - Audit of morbidity and mortality following neck of femur fracture using the POSSUM scoring system. AB - AIMS: The aims of this study were to compare the morbidity and mortality data for patients undergoing surgical fixation of a fractured neck of femur (during a 6 month period) to the predicted morbidity and mortality rates obtained from the POSSUM (Physiological and Operative Severity Score for the enUmeration of Morbidity and Mortality) scoring system, adapted for orthopaedic patients. The predictive accuracy of the orthopaedic POSSUM system is evaluated for this population. The 1-year mortality for the males and females of the study group (mean ages) is compared to the 1-year mortality of male and female New Zealanders of the same age. METHODS: Physiological and operative data was collected from patient notes; patient morbidity and mortality were obtained at 30 days and at 1 year postoperatively. The data were analysed with the orthopaedic POSSUM scoring system. RESULTS: 225 complete datasets were obtained. The mean age of the patients was 83 years; 75% were female. The observed 30-day morbidity and mortality rates were 58% and 12% respectively. The observed 1-year mortality was 38% for males (mean age 79 years) and 29% for females (mean age 84 years). New Zealand census data predicts 7% and 6.4% mortality respectively based on these mean ages. CONCLUSIONS: The POSSUM system allocates patients into groups of varying risk. The observed data shows higher numbers of complications, including death, in patients allocated into higher risk groups. The 1-year mortality is much higher than that predicted based on mean patient age from the New Zealand abridged life table. PMID- 16718300 TI - New Zealand cancer patients should have access to erythropoietin treatment. AB - Erythropoietin was the first haematopoietic growth factor to be cloned and put to clinical use in patients with anaemia. In New Zealand, the agent is approved and funded for patients with anaemia secondary to renal failure. Although there is Medsafe approval for its use in various other conditions, including cancer patients, it is not funded by Pharmac for these conditions. There is good evidence that erythropoietin will induce meaningful increases in haemoglobin levels in approximately 60% of cancer patients. Non-responders can be identified within 2-4 weeks of starting therapy and the drug discontinued, thus improving the cost-effectiveness of the programme. Responders have a significant reduction in blood transfusions, have an improved quality of life, and possibly have a better tumour response to chemo-radiotherapy, thus resulting in longer survival. British and American guidelines advocate a significantly greater use of erythropoietin than the very restrictive New Zealand use. Pharmac should review the current evidence as to the benefits of increased erythropoietin usage in New Zealand, and endorse increased access to this agent. PMID- 16718301 TI - Regarding New Zealand Medical Association's position on the minimum purchase age for alcohol. PMID- 16718302 TI - Evaluating fidelity: predictive validity for a measure of competent adherence to the Oregon model of parent management training. AB - When efficacious interventions are implemented in real-world conditions, it is important to evaluate whether or not the programs are practiced as intended. This article presents the Fidelity of Implementation Rating System (FIMP), an observation-based measure assessing competent adherence to the Oregon model of Parent Management Training (PMTO). FIMP evaluates 5 dimensions of competent adherence to PMTO (i.e., knowledge, structure, teaching skill, clinical skill, and overall effectiveness) specified in the intervention model. Predictive validity for FIMP was evaluated with a subsample of stepfamilies participating in a preventive PMTO intervention. As hypothesized, high FIMP ratings predicted change in observed parenting practices from baseline to 12 months. The rigor and scope of adherence measures are discussed. PMID- 16718303 TI - An efficacious theory-based intervention for stepfamilies. AB - This article evaluates the efficacy of the Oregon model of Parent Management Training (PMTO) in the stepfamily context. Sixty-seven of 110 participants in the Marriage and Parenting in Stepfamilies (MAPS) program received a PMTO-based intervention. Participants in the randomly assigned experimental group displayed a large effect in benefits to effective parenting practices with resultant decreases in child noncompliance and in home and school problem behaviors. MAPS findings replicate and extend those of the Oregon Divorce Study with single mothers while employing a similar multiple-method, multiple-measure randomized design, with Intent-to-Treat and structural equation modeling analyses. We discuss direct and indirect effects on distal outcomes and the generalizability of PMTO to differing family contexts. PMID- 16718304 TI - Attention affects the recognition of briefly presented visual stimuli in infants: an ERP study. AB - This study examined the effect of attention in infants on the ERP changes occurring during the recognition of briefly presented visual stimuli. Infants at ages 4.5, 6 and 7.5 months were presented with a Sesame Street movie that elicited periods of attention and inattention, and computer-generated stimuli were presented overlaid on the movie for 500 ms. One stimulus was familiar to the infants and was presented frequently, a second stimulus was familiar but presented infrequently, and a set of 14 novel stimuli were presented infrequently. An ERP component labeled the 'Nc' (Negative Central, about 450-550 ms after stimulus onset) was larger during attention than inattention and increased in magnitude over the three testing ages during attention. Late slow waves in the ERP (from 1000 to 2000 ms post-stimulus onset) consisted of a positive slow wave in response to the infrequent familiar stimulus at all three testing ages. The late slow wave in response to the infrequent novel stimulus during attention was a positive slow wave for the 4.5-month-old infants, to a positive-negative slow wave for the 6-month-old infants and a negative slow wave for the 7.5-month-old infants. These results show attention facilitates the brain response during infant recognition memory and show that developmental changes in recognition memory are closely related to changes in attention. PMID- 16718305 TI - Physiological and Emotional Reactivity to Learning and Frustration. AB - This study examined the behavioral (arm, facial) autonomic (heart rate, respiratory sinus arrhythmia [RSA], and adrenocortical axis) reactivity of 56 4 month-old infants in response to contingency learning and extinction-induced frustration. During learning, infants displayed increases in operant arm response and positive emotional expressions. Changes in average RSA(V(NA)) paralleled the observed changes in facial expressions in general and maintained an inverse relation with heart rate throughout most of the session. When frustrated by extinction, infants displayed increases in negative expressions, heart rate, and a brief increase in RSA(V(NA)) followed by a significant decrease. No significant changes were observed for cortisol. These behavioral and facial responses are consistent with earlier work. The physiological changes, along with the facial expressions and instrumental responses, indicate that the autonomic nervous system functions as a coordinated affect system by 4 months of age. PMID- 16718306 TI - Characterization of microsatellite markers in the tsetse fly, Glossina pallidipes (Diptera: Glossinidae). AB - Glossina pallidipes is a vector of African trypanosomiasis. Here we characterize eight new polymorphic microsatellite loci in 288 G. pallidipes sampled from 12 Kenya populations. The number of alleles per locus ranged from four to 36 with a mean of 20.5 +/- 10.1. Expected single locus heterozygosities varied from 0.044 to 0.829. Heterozygosity averaged 0.616 +/- 0.246. No linkage disequilibrium was found. We also report results in eight other tsetse species estimated by using the primers developed in G. pallidipes. The primers worked best in G. swynnertoni and G. austeni and worst in G. m. morsitans and G. m. submorsitans. PMID- 16718307 TI - Recent progress in cobalt-mediated [2 + 2 + 2] cycloaddition reactions. AB - For many years, our research group has been interested in the new developments of cobalt-mediated cyclizations. In this article, our recent achievements in the field of inter- and intramolecular [2 + 2 + 2] cyclizations are compiled. PMID- 16718308 TI - Smart amphiphiles: hydro/organogelators for in situ reduction of gold. AB - New urea containing gels have been used to prepare and stabilise gold nanoparticles by in situ reduction. PMID- 16718309 TI - Base-catalysed asymmetric hydroamination/cyclisation of aminoalkenes utilising a dimeric chiral diamidobinaphthyl dilithium salt. AB - A dimeric proline derived diamidobinaphthyl dilithium salt represents the first example of a chiral main group metal based catalyst for asymmetric hydroamination/cyclisation reactions of aminoalkenes. PMID- 16718310 TI - Highly diastereoselective amplification from a dynamic combinatorial library of macrocyclic oligoimines. AB - Cadmium promoted diastereoselective amplification of a single member from a dynamic combinatorial library of stereoisomeric oligoimines of different sizes allows the efficient preparation of a new macrocyclic polyamine. PMID- 16718311 TI - An azido-metal-isonicotinate complex showing long-range ordered ferromagnetic interaction: synthesis, structure and magnetic properties. AB - A new 3D Cu(II) complex [Cu1.5(N3)2(isonicotinate)]n [1], which features two types of bridging modes for azide (mu(1,1) and the rare asymmetric mu(1,1,3)) where the three bonds of the mu(1,1,3)-N3(-) group to Cu exhibit three different distances, has been synthesized and characterized, and magnetic measurements indicate that [1] experiences long-range ferromagnetic ordering at approximately 6 K. PMID- 16718312 TI - Selective aerobic oxidation in supercritical carbon dioxide catalyzed by the H5PV2Mo10O40 polyoxometalate. AB - Selective aerobic oxidation of benzylic alcohols and of activated aromatic hydrocarbons occurs in supercritical CO2 as reaction medium using H5PV2Mo10O40 as a quasi-heterogeneous catalyst without further additives or co-solvents; efficient recycling is possible and no metal leaching is detectable in the product stream. PMID- 16718313 TI - Monitoring the formation of TTF dimers by Na+ complexation. AB - The formation of the two dimeric species [(TTF)2]+* and (TTF+*)2 can be monitored by complexation of Na+ on a calix[4]arene-TTF assembly. PMID- 16718314 TI - Low pressure Pd-catalyzed carbonylation in an ionic liquid using a multiphase microflow system. AB - A low pressure microflow system was developed for palladium-catalyzed multiphase carbonylation reactions in an ionic liquid. The microflow system resulted in superior selectivity and higher yields in carbonylative Sonogashira coupling and amidation reactions of aryl iodides compared to the conventional batch system. PMID- 16718316 TI - Assessment of ordered and complementary pore volumes in polymer-templated mesoporous silicas and organosilicas. AB - A method to determine the volumes of ordered mesopores and complementary small pores in polymer-templated ordered mesoporous silicas and organosilicas is proposed on the basis of the existing relation between the pore width and unit cell values obtained by the XRD structure modeling and the adsorption pore volume. PMID- 16718315 TI - Sensory hybrid host materials for the selective chromo-fluorogenic detection of biogenic amines. AB - Pyrylium-containing mesoporous materials have been used for the chromo fluorogenic sensing of biogenic amines in an aqueous environment. PMID- 16718317 TI - A novel organic chromophore for dye-sensitized nanostructured solar cells. AB - A novel and efficient polyene-diphenylaniline dye for dye-sensitized solar cells has been synthesized. The dye has a short synthesis route and is readily adsorbed on TiO2 under a variety of dye-bath conditions. The overall solar-to-energy conversion efficiency is over 5% in the preliminary tests, in comparison with the conventional N719 dye which gives 6% under the same conditions. The dye is designed for future use also in solid state devices, with triarylamine based hole conductors. PMID- 16718318 TI - Construction of superhydrophobic surfaces by fibrous aggregation of perfluoroalkyl chain-containing organogelators. AB - Superhydrophobic surfaces, characterized by water contact angles greater than 150 degrees, can be produced by means of intermediate organogels, which were formed by perfluoroalkyl chain-containing organogelators with volatile organic solvents. PMID- 16718319 TI - Ferrimagnetic Mn2SnO4 nanowires. AB - Single-crystalline Mn2SnO4 nanowires were first synthesized by chemical vapor deposition; they consist of inverse spinel structure grown with the [111] direction; the nanowires have a ferrimagnetic phase below 46 K (T(C)) with large hysteresis; this ferrimagnetic transition is probably due to the presence of Mn3+ ions at octahedral sites. PMID- 16718320 TI - Effect of tetrabutylphosphonium cation on the physico-chemical properties of amino-acid ionic liquids. AB - Tetraalkylphosphonium-based amino-acid ionic liquids show lower viscosities and higher decomposition temperatures (>300 degrees C) than previously reported ammonium-based amino-acid ionic liquids. PMID- 16718321 TI - A novel mediatorless microbial fuel cell based on direct biocatalysis of Escherichia coli. AB - A mediatorless microbial fuel cell based on the direct biocatalysis of Escherichia coli shows significantly enhanced performance by using bacteria electrochemically-evolved in fuel cell environments through a natural selection process and a carbon/PTFE composite anode with an optimized PTFE content. PMID- 16718322 TI - A platinum-catalyzed annulation reaction leading to medium-sized rings. AB - A platinum-catalyzed domino process with intermediate benzopyrylium cations reaches its optimum utility in the formation of 7- and 8-membered rings. With iron(III) chloride, a tetracyclic product is isolated, derived from an oxidative transformation of a metal-carbene intermediate. PMID- 16718323 TI - Easy access to the family of thiazole N-oxides using HOF x CH3CN. AB - An efficient procedure for transferring an oxygen atom to thiazole-containing compounds, resulting in the corresponding N-oxides, was developed by using HOF x CH3CN; mild reaction conditions, high yields and easy purification are the main features of this novel route, while X-ray structural analysis reveals a hydrogen bond between the N-oxide functionality and a water molecule. PMID- 16718324 TI - Fabrication of bismuth subcarbonate nanotube arrays from bismuth citrate. AB - Bismuth subcarbonate ((BiO)2CO3) nanotubes with uniform diameters of about 3-5 nm were fabricated from bismuth citrate; these nanotubes exhibit antibacterial properties against Helicobacter pylori (50% inhibition at 10 microg mL(-1)), a bacterium causing peptic ulcers and gastritis. PMID- 16718325 TI - Scanning electrochemical microscopy under illumination: an elegant tool to directly determine the mobility of charge carriers within dye-sensitized nanostructured semiconductors. AB - The diffusion constant of the ferrocenium ion in dye-sensitized nanostructured materials has been determined by time-of-flight experiments under working solar cell conditions with scanning electrochemical microscopy. PMID- 16718326 TI - Synthesis of pyrroles: reaction of chromium N-alkylaminocarbene complexes with alpha,beta-unsaturated aldehydes. AB - N-alkylaminocarbene complexes of chromium were found to react with alpha,beta unsaturated aldehydes to give pyrroles in good yields. PMID- 16718327 TI - Bergman cyclopolymerization within the channels of functional hybrid nanocomposites formed by co-assembly of silica and polymerizable surfactant monomer. AB - The Bergman cyclopolymerization of polymerizable surfactant monomer was carried out within the hexagonal channels of functional hybrid nanocomposite formed by co assembly with silica. PMID- 16718328 TI - Characterisation of the thermally accessible spin triplet state in dimers of the 7pi ClCNSSS+* in the solid state. AB - [ClCNSSS]2(2+) is the first example of a thiazyl radical dimer where population of a thermally excited spin triplet state has been detected, as is proved by VT powder and single-crystal EPR spectroscopy. PMID- 16718330 TI - A star-shaped ruthenium complex with five ferrocenyl-terminated arms bridged by trans-platinum fragments. AB - We present the synthesis of the new heteropolytopic penta(4 ethynylphenyl)cyclopentadiene ligand, its complexation through the Cp ring to ruthenium tris(indazolyl)borate and through the terminal alkyne groups to five ferrocenyl ethynyl platinum units, yielding an undecanuclear heterotrimetallic complex. PMID- 16718329 TI - Desymmetrization of diols by a tandem oxidation/Wittig olefination reaction. AB - Diols are desymmetrized by a tandem oxidation/Wittig olefination to give alpha,beta-unsaturated hydroxy esters without the requirement for protecting group strategies; the alpha,beta-unsaturated hydroxy esters are transformed into dienyl diesters using a second oxidation/Wittig olefination sequence using PCC. PMID- 16718331 TI - Instant nano-hydroxyapatite: a continuous and rapid hydrothermal synthesis. AB - Nano-particle hydroxyapatite (HA) rods, were rapidly synthesised using a three pump continuous hydrothermal process (using a water feed at up to 400 degrees C and at 24 MPa): the product was obtained as a highly crystalline and phase pure material, without the need for an ageing step or subsequent heat treatment. PMID- 16718332 TI - A tripodal sulfur ligand for the selective ruthenium-catalysed hydrogenation of dimethyl oxalate. AB - The first example of a catalyst utilising a sulfur-based ligand [MeC(CH2SBu)3] for the selective hydrogenation of dimethyl oxalate to methyl glycolate is reported. PMID- 16718333 TI - Chiral diphosphites and diphosphoramidites as cheap and efficient ligands in Rh catalyzed asymmetric olefin hydrogenation. AB - Chiral diphosphites and diphosphoramidites derived from BINOL or diphenylprolinol are efficient ligands in asymmetric Rh-catalyzed olefin hydrogenation, provided the proper achiral backbone is chosen. PMID- 16718334 TI - The vitality of uranium molecular chemistry at the dawn of the XXIst century. AB - The intent of this Dalton Perspective is to highlight the recent advances in uranium molecular chemistry, with the results reported during the 2000-2006 period. This discipline is currently witnessing an impressive development, together with the theoretical chemistry and solid-state chemistry of the f elements, and its face has profoundly changed, revealing unsuspected structural and reactivity features. This progress required and was facilitated by the use of new precursors. Studies of low-valent compounds gave a better insight into lanthanide(III)/actinide(III) differentiation and led to the discovery of unusual reactions, including activation of small molecules. A number of tetravalent uranium complexes, in particular polynuclear compounds, have been synthesized, which exhibit exciting structures and physicochemical properties. The potential of uranium(III) and uranium(IV) complexes in catalysis has been confirmed. The uranyl complexes, from mononuclear species to supramolecular assemblies, reveal a variety of novel structures, changing the generally accepted ideas on the coordination geometry and the stability of the UO2(2+) ion. PMID- 16718335 TI - Structural, spectroscopic, and electrochemical behavior of trans-phenolato cobalt(III) complexes of asymmetric NN'O ligands as archetypes for metallomesogens. AB - In order to understand and predict structural, redox, magnetic, and optical properties of more complex and potentially mesogenic electroactive compounds such as [Co(III)(L(t-BuLC))2]ClO4 (1), five archetypical complexes of general formula [Co(III)(L(RA))2]ClO4, where R = H (2), tert-butyl (3), methoxy (4), nitro (5), and chloro (6), were obtained and studied by means of several spectrometric, spectroscopic, and electrochemical methods. The complexes 2, 4, and 6 were characterized by single-crystal X-ray diffraction, and show the metal center in an approximate D2h symmetry. Experimental results support the fact that the electron donating or withdrawing nature of the phenolate-appended substituents changes dramatically the redox and spectroscopic properties of these compounds. The 3d6 electronic configuration of the metal ion dominates the overall geometry adopted by these compounds with the phenolate rings occupying trans positions to one another. Formation of phenoxyl radicals has been observed for 1, 3, and 6, but irreversible ligand oxidation takes place upon bulk electrolysis. These data were compared to detailed B3LYP/6-31G (d)-level computational calculations and have been used to account for the results observed. A comparison between compound 1 and archetype 3, validates the approach of using archetypical models to study metal-containing soft materials. PMID- 16718336 TI - Influence of gamma irradiation on hydrophobic room-temperature ionic liquids [BuMeIm]PF6 and [BuMeIm](CF3SO2)2N. AB - Stability of neat hydrophobic Room-Temperature Ionic Liquids (RTIL) [BuMeIm]X, where [BuMeIm]+ is 1-butyl-3-methylimidazolium and X- is PF6-, and (CF3SO2)2N-, was studied under gamma radiolysis (137Cs) in an argon atmosphere and in air. It was found that the density, surface tension, and refraction index of RTILs are unchanged even by an absorbed dose of approximately 600 kGy. Studied RTILs exhibit considerable darkening when subjected to gamma irradiation. The light absorbance of ionic liquids increases linearly with the irradiation dose. Water has no influence on radiolytic darkening. A comparative study of [BuMeIm]X and [Bu4N][Tf2N] leads to the conclusion that the formation of colored products is related to gamma radiolysis of the [BuMeIm]+ cation. The radiolytic darkening kinetics of RTILs is influenced by the anions as follows: Cl- < (CF3SO2)2N- < PF6 . Electrospray ionization mass spectrometry and NMR analysis reveal the presence of nonvolatile radiolysis products at concentrations below 1 mol% for an absorbed dose exceeding 1200 kGy. Initial step of BuMeIm+ cation radiolysis is the loss of the Bu* group, the H* atom from the 2 position on the imidazolium ring, and the H* atom from the butyl chain. Radiolysis of ionic liquid anions yields F* and CF3* from PF6- and [Tf2N]-, respectively. Recombinations of these primary products of radiolysis lead to various polymeric and acidic species. PMID- 16718337 TI - Migratory insertion in N-heterocyclic carbene-containing Fe carbonyl complexes: an experimental and theoretical study. AB - The compound [Fe(eta-C5H5)(CO)2(Me)] reacts thermally with N-heterocyclic carbenes (L) to give both alkyl, [Fe(eta-C5H5)(L)(CO)(Me)], and acyl, [Fe(eta C5H5)(L)(CO)(COMe)], derivatives. The reaction temperature has been shown to affect the product distribution. The alkyl and acyl derivatives exist in an equilibrium that is more easily perturbed than in the tertiary phosphine analogues. DFT studies on the reactivity of [Fe(eta-C5H5)(CO)2(Me)] with PH3 and dihydroimidazole-2-ylidene (IH) have shown that CO exchange is energetically favoured for IH, and energetically disfavoured for PH3. The products of CO induced migratory insertion, [Fe(eta-C5H5)(L)(CO)(COMe)], are more stable than the parent alkyl, [Fe(eta-C5H5)(L)(CO)(Me)], compounds. This stabilisation is larger when L = IH than when L = PH3. Stabilisation of the transition state by agostic interactions was seen in both instances, but this was significantly more pronounced for L = IH. PMID- 16718339 TI - Platinum(II) complexes bearing 1,1'-bis(diphenylphosphino)ferrocene as building blocks for functionalized redox active porphyrins. AB - Reactions of the cationic complex ions [PtMe(Me2SO)(PP)]+ (PP = dppf (1,1' bis(diphenylphosphino)ferrocene) and dppe (1,2-bis(diphenylphosphino)ethane)) with 5,10,15,20-tetrakis(4-pyridyl)-21H,23H-porphyrin (TpyP) led to the formation of the symmetrical tetraplatinated porphyrin complexes, [PtMe(PP)]4TpyP.X4 (PP = dppf, X = CF3SO3-, 3, and PP = dppe, X = BF4-, 5) containing the organometallic fragment [PtMe(PP)]. The precursor sulfoxide platinum complexes [PtMe(Me2SO)(dppf)]CF3SO3, 2 and [PtMe(Me2SO)(dppe)]BF4, 4, were prepared by halide abstraction from [PtMeCl(dppf)], 1, and by controlled protonolysis of [PtMe2(dppe)] respectively, in the presence of a small amount of dimethyl sulfoxide. All these starting platinum(II) compounds, as well as the porphyrin derivatives 3 and 5, were fully characterized through elemental analysis, 1H NMR mono- and bidimensional, 31P[1H], 31P-1H HMBC, UV/Vis absorption and photophysical measurements. The X-ray crystal structure of complex 1 has been determined. In order to ascertain the electronic influence of ferrocene, the spectroscopic and redox properties of 3 were compared with those of TPyP and of the analogous 5. Cyclic voltammetry (CV), differential pulse voltammetry (DPV), 1H and 31P NMR data, and UV/Vis data, all suggest a certain degree of communication between the central porphyrin and the peripheral hetero-bimetallic fragments. In contrast, no detectable interaction among these peripheral groups seem to come into play. Unlikely from the porphyrin derivative 5, formation of well defined fluorescent mesoscopic ring structures was easily achieved by simple evaporation from diluted dichloromethane solutions of 3. PMID- 16718338 TI - Synthesis, a case of isostructural packing, and antimicrobial activity of silver(I)quinoxaline nitrate, silver(I)(2,5-dimethylpyrazine) nitrate and two related silver aminopyridine compounds. AB - The synthesis and low temperature crystal structures of [Ag(quinoxaline)]n(NO3)n, 1, [Ag(2,5-dimethylpyrazine)(NO3)]n, 2 and [Ag4(3-aminopyridine)4(NO3)4]n 3 are presented. The quinoxaline compound forms a 1D coordination polymer with the characteristic linear 2-coordination figure of silver(I), the N-Ag-N angle being 164.2(1) degrees, and only weak silver-nitrate interactions. In addition there is an interaction giving pairs of parallel chains as the main structural theme. The 2,5-dimethylpyrazine compound has approximately trigonal-planar coordination, also binding one nitrate at the relatively short Ag-O distances 2.444(3) angstroms and 2.484(3) angstroms, respectively, for the two crystallographically different silver atoms. This also results in a 1D coordination polymer that, despite the large differences in the Ag(I) coordination environment, is isostructural with 1. [Ag4(3-aminopyridine)4(NO3)4]n 3 forms a 2D coordination polymer by bridging nitrate ions. The antimicrobial activity of 1-3, and also of [Ag3(2-aminopyridine)4](NO3)3, 4 was screened for 13 different pathogens and substantial activity was shown for 1 against Escherichia coli and Pseudomonas aeruginosa (MIC 4 microg cm(-3)) and somewhat lower activity was registered against Sarcina lutea and Salmonella typhi for 1, Bordetella bronchiseptica for 2, Salmonella typhi and Pseudomonas aeruginosa for 3, and Escherichia coli and Shigella sonnie for 3 (MIC 8 microg cm(-3)). Only low activity was shown against the yeast Candida albicans for 1, 2 and 4 whereas no activity against this pathogen was registered for 3. PMID- 16718340 TI - Synthesis, structures and reactions of cyclometallated gold complexes containing (2-diphenylarsino-n-methyl)phenyl (n = 5, 6). AB - Reaction of (C6H3-2-AsPh2-n-Me)Li (n = 5 or 6) with [AuBr(AsPh3)] at -78 degrees C gives the corresponding cyclometallated gold(I) complexes [Au2[(mu-C6H3-n Me)AsPh2]2] [n = 5, (1); n = 6, (9)]. 1 undergoes oxidative addition with halogens and with dibenzoyl peroxide to give digold(II) complexes [Au2X2[(mu-C6H3 5-Me)AsPh2]2] [X = Cl (2a), Br (2b), I (2c) and O2CPh (3)] containing a metal metal bond between the 5d9 metal centres. Reaction of 2a with AgO2CMe or of 3 with C6F5Li gives the corresponding digold(II) complexes in which X = O2CMe (4) and C6F5 (6), respectively. The Au-Au distances increase in the order 4 < 2a < 2b < 2c < 6, following the covalent binding tendency of the axial ligand. Like the analogous phosphine complexes, 2a-2c and 6 in solution rearrange to form C-C coupled digold(I) complexes [Au2X2[mu-2,2-Ph2As(5,5-Me2C6H3C6H3)AsPh2]] [X = Cl (5a), X = Br (5b), X = I (5c) and C6F5 (7)] in which the gold atoms are linearly coordinated by As and X. In contrast, the products of oxidative additions to 9 depend markedly on the halogens. Reaction of 9 with chlorine gives the gold(I) gold(III) complex, [ClAu[mu-2-Ph2As(C6H3-6-Me)]AuCl[(6-MeC6H3)-2-AsPh2] kappa2As,C] (10), which contains a four-membered chelate ring, Ph2As(C6H3-6-Me), in the coordination sphere of the gold(III) atom. When 10 is heated, the ring is cleaved, the product being the digold(I) complex [ClAu[mu-2-Ph2As(C6H3-6 Me)]Au[AsPh2(2-Cl-3-Me-C6H3)]] (11). Reaction of 9 with bromine at 50 degrees C gives a monobromo digold(I) complex (12), which is similar to 11 except that the 2-position of the substituted aromatic ring bears hydrogen instead halogen. Reaction of 9 with iodine gives a mixture of a free tertiary arsine, (2-I-3 MeC6H3)AsPh2 (13), a digold diiodo compound (14) analogous to 11, and a gold(I) gold(III) zwitterionic complex [I2Au(III)[(mu-C6H3-2-AsPh2-6-Me)]2Au(I)] (15) in which the bridging units are arranged head-to-head between the metal atoms. The structures of 2a-2c and 4-15 have been determined by single-crystal X-ray diffraction analysis. The different behaviour of 1 and 9 toward halogens mirrors that of their phosphine analogues; the 6-methyl substituent blocks C-C coupling of the aryl residues in the initially formed oxidative addition product. In the case of 9, the greater lability of the Au-As bond in the initial oxidative addition product may account for the more complex behaviour of this system compared with that of its phosphine analogue. PMID- 16718341 TI - Chiral aggregation and spontaneous resolution of thiosemicarbazone metal complexes. AB - Chiral aggregation and spontaneous resolution of thiosemicarbazone metal complexes MnL2 (1 and 1') (HL = acetylpyrazine thiosemicarbazone) were achieved through cooperation of hydrogen bonding and pi-pi stacking interactions. Compound 1 crystallized in a chiral space group P4(1)2(1)2 and the molecules exhibited lambda-configuration. Head-to-tail pyrazine-amino hydrogen bonding linked the molecules together presenting a two-dimensional homochiral sheet. The interlayer pi-pi stacking interactions linked the layers in a homochiral array and extended the 2D homochirality to a three-dimensional network. Compound 1 crystallized in space group P4(3)2(1)2, the opposite handedness of P4(1)2(1)2. The molecules, the two-dimensional hydrogen-bonded sheets and the three-dimensional network all exhibited the opposite chiralities to those of 1. Solid state CD spectra measurements confirmed the occurrence of spontaneous resolution of MnL2 and demonstrated that one of the enantiomers of the compound was always in excess. The zinc complexes (2 and 3) and nickel complex 4 crystallized in the chiral space group P4(3)2(1)2. Similar two-dimensional pyrazine-amine hydrogen bonded chiral sheets and three-dimensional networks were found in the crystal packing patterns. The cadmium complex 5 crystallized in a chiral space group P2(1)2(1)2(1), however, it exhibited a similar chiral packing pattern. The only difference was that enlarged Cd-S and Cd-N bond lengths induced the potential 4(1) symmetry. All results indicated that the supramolecular synthon, the cooperation of the pyrazine-amino hydrogen bonding and the pi-pi stacking interactions, were robust enough so that the orientation of the molecules in the solid state could be predicted to a reasonable degree of accuracy. PMID- 16718342 TI - Synthesis, molecular structure and norbornene polymerization behavior of three coordinate nickel(I) complexes with chelating anilido-imine ligands. AB - Reaction of lithium salts of anilido-imine ligands bearing bulky substituentes on the nitrogen donor atoms with trans chloro(phenyl)bis(triphenylphosphane)nickel(II) results in the formation of two rare three-coordinate nickel(I) complexes [(Ar1N=CHC6H4NAr2)Ni(I)PPh3] (1: Ar1 = Ar2 = 2,6-i-Pr2C6H3; 2: Ar1 = 2,6-Me2C6H3, Ar2 = 2,6-i-Pr2C6H3). The molecular structures of complexes 1 and 2 have been confirmed by single crystal X-ray analyses. These two complexes exhibit paramagnetic properties as measured by their EPR and 1H NMR spectra. After being activated with methylaluminoxane (MAO) these complexes could polymerize norbornene to afford addition-type polynorbornene (PNB) with high molecular weight M(w) (10(6) g mol(-1)), catalytic activities being high, up to 2.82 x 10(7) g(PNB) mol(-1)(Ni) h(-1). PMID- 16718343 TI - Synthesis and characterisation of four- and six-membered P-Se heterocycles. AB - The preparation, spectroscopic characterisation and crystal structures of [FcP(mu Se)Se]2, [FcP(mu-Se2)Se]2 and [PhP(mu-Se2)Se]2 are reported. Crystallographic data reveal planar four-membered PSePSe and skewed six-membered P2Se4 rings, respectively, in all cases with trans arrangement of organic substituents and exo selenium atoms. Whilst stable at room temperature in solid state, NMR data suggest the six-membered rings of both the ferrocenyl and phenyl compounds decompose in the solution with loss of red selenium, forming PSe2PSe five membered rings. PMID- 16718344 TI - An electrochemical and DFT study on selected beta-diketiminato metal complexes. AB - Selected homoleptic metal beta-diketiminates M(I)L and M(II)L2 [M(I) = Li or K, M(II) = Mg, Ca or Yb; L: L(Ph) = [N(SiMe3)C(Ph)]2CH, L(Bu(t)) = N(SiMe3)C(Ph)C(H)C(Bu(t))N(SiMe3), L* = [N(C6H3Pr(i)2-2,6)C(Me)]2CH] have been studied by cyclic voltammetry (CV). The primary reduction (E(p)red, the peak reduction potential measured vs. SCE in thf containing 0.2 M [NBu4][PF6] with a scan rate 100 mV s(-1) at a vitreous carbon electrode at ambient temperature) is essentially ligand-centred: E(p)red being ca. -2.2 V (LiL(Ph) and KL(Ph)) and 2.4 V [Mg(L(Ph))2, LiL(Bu(t)) and Ca(L(Ph))2], while LiL* is significantly more resistant to reduction (E(p)red = -3.1 V). These observations are consistent with the view that the two (L(Ph)) or single (L(Bu(t))) C-phenyl substituent(s), respectively, are available for -electron-delocalisation of the reduced species, whereas the N-aryl substituents of L* are unable to participate in such conjugation for steric reasons. The primary reduction process was reversible on the CV-time scale only for LiL(Bu(t)), Ca(L(Ph))2 and Yb(L(Ph))2. For the latter this occurs at a potential ca. 500 mV positive of Ca(L(Ph))2, consistent with the notion that the LUMO of Yb(L(Ph))2 has substantial metal character. The successive reversible steps, each separated by ca. 500 mV, indicate that there is strong electronic communication between the two ligands of Yb(L(Ph))2. The overall three-electron transfer sequence shows that the final reduction level corresponds to [Yb(II)(L(Ph))2-(L(Ph))3-]. DFT calculations on complexes Li(L(Ph))(OMe2)2 and Li2(L(Ph))(OMe2)3 showed that both HOMO and LUMO orbitals are only based on the ligand with a HOMO-LUMO gap of 4.21 eV. Similar calculations on a doubly reduced complex Yb[(mu-L(Ph))Li(OMe2)]2 demonstrated that there is a considerable Yb atomic orbital contribution to the HOMO and LUMO of the complex. PMID- 16718345 TI - Preparation of five- and six-coordinate aryl(hydrido) iridium(III) complexes from benzene and functionalized arenes by C-H activation. AB - The reaction of the in situ generated cyclooctene iridium(I) derivative trans [IrCl(C8H14)(PiPr3)2] with benzene at 80 degrees C gave a mixture of the five coordinate dihydrido and hydrido(phenyl) iridium(III) complexes [IrH2(Cl)(PiPr3)2] 2 and [IrH(C6H5)(Cl)(PiPr3)2] 3 in the ratio of about 1 : 2. The chloro- and fluoro-substituted arenes C6H5X (X = Cl, F), C6H4F2 and C6H4F(CH3) reacted also by C-H activation to afford the corresponding aryl(hydrido) iridium(III) derivatives [IrH(C6H4X)(Cl)(PiPr3)2] 7, 8, [IrH(C6H3F2)(Cl)(PiPr3)2] 9-11 and [IrH[C6H3F(CH3)](Cl)(PiPr3)2] 12, 13, respectively. The formation of isomeric mixtures had been detected by 1H, 13C, 19F and 31P NMR spectroscopy. Treatment of 3 and 7-13 with CO gave the octahedral carbonyl iridium(III) complexes [IrH(C6H3XX')(Cl)(CO)(PiPr3)2] 5, 14-20 without the elimination of the arene. The reactions of trans-[IrCl(C8H14)(PiPr3)2] with aryl ketones C6H5C(O)R (R = Me, Ph), aryl ketoximes C6H5C(NOH)R (R = Me, Ph) and benzaloxime C6H5C(NOH)H resulted in the formation of six-coordinate aryl(hydrido) iridium(III) compounds 21-25 with the aryl ligand coordinated in a bidentate kappa2-C,O or kappa2-C,N fashion. With C6H5C(O)NH2 as the substrate, the two isomers [IrH[kappa2-N,O-NHC(O)C6H5](Cl)(PiPr3)2] 26 and [IrH[kappa2-C,O C6H4C(O)NH2](Cl)(PiPr3)2] 27 were prepared stepwise. Treatment of trans [IrCl(C8H14)(PiPr3)2] with benzoic acid gave the benzoato(hydrido) complex [IrH[kappa2-O,O-O2CC6H5](Cl)(PiPr3)2] 29 which did not rearrange to the kappa2 C,O isomer. PMID- 16718346 TI - Neuropathology of Alzheimer disease: pathognomonic but not pathogenic. AB - Neuropathological changes in subjects with dementia are, by definition, end-stage phenomena. While such changes allow case characterization and lend themselves to disease classification and modeling, the lesions themselves are not etiological. This truth would appear to be self-evident, yet the medical and scientific literature suggests otherwise. Indeed it is now customary to view amyloid plaques in Alzheimer disease as primary etiological, neurotoxic lesions and, hence, removing them (e.g., by immunotherapy) is believed to lead to clinical improvement. The foundation for this line of thinking lies in the existence of rare kindreds with mutations in amyloid-beta, or mutations believed to be involved in the processing of amyloid-beta, and then the extrapolation of the inherited condition to sporadic disease. We believe that this overall construct ignores early events that are more critical to onset and progression of sporadic disease. Likewise, we have studied subjects with sporadic Alzheimer disease, as well as early onset familial Alzheimer disease and Down's syndrome, over a spectrum of ages, and have found that markers of oxidative stress precede amyloid deposits in all three conditions. Amyloid and neurofibrillary pathology in the Alzheimer brain show a decrease in oxidative stress relative to vulnerable but morphologically intact neurons, suggesting that neurodegenerative lesions are compensatory phenomena, and thus manifestations of cellular adaptation. The pathology of neurodegenerative diseases should be viewed as the end-stage consequence, as opposed to cause, of the disease processes, so that early disease processes that are amenable to intervention can be properly recognized and treated. PMID- 16718347 TI - Relative paucity of tau accumulation in the small areas with abundant Abeta42 positive capillary amyloid angiopathy within a given cortical region in the brain of patients with Alzheimer pathology. AB - Cerebral amyloid angiopathy (CAA) is a manifestation of amyloid beta-protein (Abeta) accumulation in the elderly as well as in patients with Alzheimer's disease (AD). Two types of CAA have been noted, based on the type of vasculature in which Abeta is deposited: cerebral capillary amyloid angiopathy (capCAA) and non-capCAA. Non-capCAA is a common form of CAA that consists of Abeta deposited in arteries and arterioles. Recent information on Abeta metabolism in the brain suggests that non-capCAA is associated with Abeta secretion into the cerebrospinal fluid via the perivascular space, whereas capCAA is associated with Abeta removal to blood plasma via the capillary endothelium. Abeta40, a major and relatively soluble Abeta isoform, is deposited predominantly in non-capCAA, and Abeta42, which is insoluble and associated more closely than Abeta40 with AD, is deposited predominantly in capCAA. Studying small areas of microscopic size within a given cortical region, we found an inverse association of capCAA and senile plaques. We also found a relative paucity of tau pathology in the small areas with abundant capCAA compared with the small areas with abundant senile plaques within a cortical region with the same cytoarchitecture. We suppose that both capCAA and senile plaques indicate high Abeta42 in the neuropil but that only Abeta42 in the form of insoluble deposits in senile plaques promotes tau abnormality. PMID- 16718348 TI - Canine cognitive deficit correlates with diffuse plaque maturation and S100beta ( ) astrocytosis but not with insulin cerebrospinal fluid level. AB - Like humans, canines develop with aging beta-amyloid (Abeta) plaques and a progressive cognitive deficit on tasks similar to those used in diagnosis and follow-up of Alzheimer's disease. Owing to that, dogs are quite unique to investigate the early events taking place in the diffuse Abeta plaque maturation and its relationship with cognitive deficit. The aim of the present investigation was to study the link between the diffuse Abeta plaque maturation and the astro- and microglial reactivity. The involvement of insulin and beta-subunit of S100 protein (S100beta) overexpression in the process was also investigated. Abeta plaques were measured and counted in prefrontal cortex of 16 pet dogs of different breeds, weight and sex, classified as control and with a light or severe cognitive deficit. A correlation between canine graded cognitive deficit, diffuse plaque maturation, and S100beta (-) astrocytosis, but not with cerebrospinal fluid insulin level, was found that may reflect the very early events of Abeta deposition in Alzheimer's disease. PMID- 16718349 TI - Accelerated Tau deposition in the brains of individuals infected with human immunodeficiency virus-1 before and after the advent of highly active anti retroviral therapy. AB - This study aims to investigate the influence of human immunodeficiency virus (HIV) infection on the neurodegenerative processes normally associated with ageing. We have looked for evidence of beta amyloid and hyperphosphorylated Tau deposition in HIV-infected subjects before and after the advent of highly active anti-retroviral therapy (HAART). In addition we have looked for evidence of axonal damage. We have compared these HIV-positive cases with age-matched controls and with older non-demented controls. We find no evidence of significant premature beta amyloid deposition in HIV-infected cases; however, we do observe elevated levels of hyperphosphorylated Tau in the hippocampus of many HIV infected subjects, compared with age-matched controls. The greatest levels of hyperphosphorylated Tau are noted in HAART-treated subjects. Axonal damage marked by expression of beta amyloid pre-cursor protein (BAPP) was highly variable in all groups including control subjects. We surmise that HIV infection and/or the use of anti-retroviral therapy may predispose to accelerated neuroageing in the form of hyperphosphorylated Tau deposition in the hippocampus. Within the age groups studied these significant neuropathological changes remained subclinical and were not yet associated with cognitive impairment. PMID- 16718351 TI - Regulation of microglial cell responses in murine Toxoplasma encephalitis by CD200/CD200 receptor interaction. AB - Under autoimmune inflammatory conditions within the brain, evidence suggests that neurons downregulate microglial activation through CD200/CD200R interaction, which reduces disease severity. To gain insight into the regulation of intracerebral immune reactions by resident brain cells in chronic cerebral infections, the expression of the CD200 antigen and the CD200R as well as the functional role of CD200/CD200R interactions were characterized in murine Toxoplasma encephalitis. In the normal brain of C57BL/6 wild type mice, CD200 was ubiquitously expressed on neurons, their axons, cerebral endothelial cells, and plexus macrophages. CD200R was expressed at very low levels on cerebral macrophages and microglia without differences between CD200-/- and wild type mice. Infection of C57BL/6 mice with Toxoplasma gondii induced an upregulation of CD200R on microglia and of CD200 on blood vessel endothelial cells. In Toxoplasma encephalitis of CD200-/- mice, microglial cell numbers strongly increased due to an enhanced proliferation indicated by increased Ki-67 immunoreactivity. In addition, microglial activation was increased in CD200-/- mice as evidenced by a further upregulation of already high MHC class II levels as well as an increased expression of the anti-parasitic effector molecules, TNF and iNOS. The increased microglial cell activation resulted in a reduced intracerebral parasite burden and an increased survival rate. Thus, in Toxoplasma encephalitis, microglial activity was regulated via CD200/CD200R-mediated interaction further pointing to an intrinsic regulation of brain resident cells under inflammatory CNS conditions. PMID- 16718350 TI - Transient axonal injury in the absence of demyelination: a correlate of clinical disease in acute experimental autoimmune encephalomyelitis. AB - Axonal degeneration contributes to the transient and permanent neurological deficits seen in multiple sclerosis, an inflammatory disease of the central nervous system. To study the immunological mechanisms causing axonal degeneration, we induced experimental autoimmune encephalomyelitis (EAE) in wildtype Lewis rats and Lewis rats with a slowly progressive myelin degeneration due to proteolipid protein (PLP) overexpression. EAE was triggered either by the transfer of encephalitogenic T-cells alone or by the co-transfer of T-cells with demyelinating antibodies. Inducible nitric oxide synthase (iNOS) expression in perivascular macrophages was associated with a transient functional disturbance of axons, reflected by the focal and reversible accumulation of amyloid precursor protein. Clinical disease correlated with the numbers of APP positive axon spheroids. Demyelination was associated with a further increase of iNOS expression in macrophages and with a higher degree of axonal injury. Our studies suggest that nitric oxide and its metabolites contribute to axonal pathology and possibly also to subsequent neurological dysfunction in EAE. PMID- 16718352 TI - Cytogenetic features of ependymoblastomas. AB - Ependymoblastomas are very rare and highly malignant embryonal tumours of the central nervous system with distinctive multilayered rosettes being the main histopathological feature. They are a diagnostically challenging subtype of embryonal tumours, whose genetic features are unknown. Primary ependymoblastomas from four children (one boy, three girls; mean age 24.8 months, range 4-41 months) were investigated by comparative genomic hybridisation (CGH), to our knowledge constituting the only cohort of this entity studied by cytogenetic means. DNA copy number changes were found in each case, consisting mainly of gains of chromosome 2 as well as losses of chromosomes 6q and 13q (75% each). The tumours showed between one and five aberrations with a mean of 3.25 DNA copy number changes per case, with gains being less frequent than losses (1.25 gains vs 2 losses per case). The youngest patient showed the least imbalances (one), whereas the oldest child presented with the most aberrations (five). Clinical follow-up data were available for three of the four patients. All three had died of their disease after a post-operative survival of 9 months (range 6-14 months). Our CGH data suggest that ependymoblastomas show distinct and fairly consistent chromosomal aberrations. PMID- 16718353 TI - Podoplanin expression in primary central nervous system germ cell tumors: a useful histological marker for the diagnosis of germinoma. AB - Podoplanin, a mucin-like transmembrane sialoglycoprotein, promotes platelet aggregation and may be involved in cancer cell migration, invasion, metastasis, and malignant progression. Podoplanin/aggrus is highly expressed in testicular seminoma, suggesting that it may be a sensitive marker for testicular seminomas. Here we investigated the expression of podoplanin in central nervous system (CNS) germ cell tumors (GCTs) by immunohistochemical staining of tumor samples from 62 patients. In 40 of 41 (98%) germinomas (including germinomatous components in mixed GCTs), podoplanin was diffusely expressed on the surface of germinoma cells; lymphocytes, interstitial cells, and syncytiotrophoblastic giant cells were negative for podoplanin. Except for immature teratomas (12/17; 71%), podoplanin expression was absent in non-germinomatous GCTs, including seven teratomas, seven embryonal carcinomas, seven yolk sac tumors, and seven choriocarcinomas. In immature teratomas, focal podoplanin staining was observed in fewer than 10% of immature squamous and columnar epithelial cells. Thus, podoplanin expression may be a sensitive immunohistochemical marker for germinoma in CNS GCTs. As such, it may be useful for diagnosis, for monitoring the efficacy of treatment, and as a potential target for antibody-based therapy. PMID- 16718354 TI - Nitric oxide synthase in muscular dystrophies: a re-evaluation. AB - Duchenne and Becker muscular dystrophies (DMD and BMD) are associated with decreased total nitric oxide (NO). However, mechanisms leading to NO deficiency with consequent muscle-cell degeneration remain unknown. To address this issue, we examined skeletal muscles of DMD and BMD patients for co-expression of NO synthase (NOS) with nitrotyrosine and transcription factor CREB, as well as with enzymes engaged in NO signaling. Employing immunocytochemical labeling, Western blotting and RT-PCR, we found that, in contrast to the most commonly accepted view, neuronal NOS was not restricted to the sarcolemma and that muscles of DMD and BMD patients retained all three NOS isoforms with an up-regulation of the inducible NOS isoform, CREB and nitrotyrosine. We suggest that enhanced nitrotyrosine immunostaining in muscle fibers as well as in the vasculature of DMD and BMD specimens reflects massive oxidative stress, resulting in withdrawal of NO from its regular physiological course via the scavenging actions of superoxides. PMID- 16718355 TI - Functional recovery improvement is related to aberrant reinnervation trimming. A comparative study using fresh or predegenerated nerve grafts. AB - Experimentally predegenerated nerve grafts have been demonstrated to improve recovery. In a 12 month-long study, we compared the degree of recovery of conventional and predegenerated grafts in rat median nerve repair. To induce predegeneration the ulnar donor nerve was crushed and grafting to the median nerve was performed 2 weeks later. The day of recovery and the improvement of finger flexion strength were studied by the grasping test. At 3, 6, 9, and 12 months after surgery retrograde labeling studies and flexor carpi radialis muscle ATPase histochemistry were performed. In the predegenerated grafts, the recovery of finger flexion occurred 19.6+/-1.5 days after surgery and was significantly faster than that in the conventional group. Twelve months after surgery, a similar rate of 85% of grasping strength recovery in relation to the normal control rats was demonstrated for the conventional and predegenerated grafts. After grafting, a larger number of motoneurons, compared to the normal controls, were retrograde labeled in the median nerve. This surplus of retrograde labeled motoneurons in the predominantly sensory branch of the median nerve represented misdirected motor fibers. There was a time-related decrease in the number of labeled motoneurons, which correlated to functional grasping strength recovery. Muscle reinnervation induced a predominance of type I over type II muscle fibers. Forty percent of type I fibers were grouped indicating that collateral sprouting plays a prominent role during muscle reinnervation. Regeneration in predegenerated grafts was faster but the final rate of recovery was similar to conventional grafts. PMID- 16718356 TI - Co-existence of paraganglioma and myxopapillary ependymoma of the cauda equina. PMID- 16718357 TI - Complete nucleotide sequence of the genomic RNA of an Amami-O-shima strain of East Asian Passiflora potyvirus. PMID- 16718358 TI - A broader definition of 'the virus species'. AB - We propose that the formal definition of a virus species by the International Committee on Taxonomy of Viruses (ICTV) should be broadened by removing the restrictive word "polythetic" from the current definition, so that any characters can be used to define species. This change will bring the definition of virus species into line with the species definitions of cellular organisms and broaden the range of characters available for describing virus species. PMID- 16718359 TI - Tissue-specific and fatty acid transporter-specific changes in heart and soleus muscle over a 1-yr period. AB - Rates of fatty acid oxidation increase rapidly in both rat heart and skeletal muscle in the early postnatal period. Therefore, we examined in heart and soleus muscle, (a) whether there were rapid changes in fatty acid transporter (FAT/CD36, FABPpm) mRNA and protein expression early in life (days 10 -36) and thereafter (days 84, 160, 365), and (b) whether the rates of fatty acid transport and the plasmalemmal content of FAT/CD36 and FABPpm were altered. Protein expression was altered rapidly from day 10-36 in both heart (FAT/CD36 only, +21%, P < 0.05)) and soleus muscle (FAT/CD36 + 100%, P < 0.05; FABPpm -20%, P < 0.05), with no further changes thereafter (P < 0.05). Rates of fatty acid transport (day 10 vs day 160) were increased in heart (+33%, P < 0.05) and muscle (+85%, P < 0.05), and were associated with concomitant increases in plasmalemmal FABPpm (+44%, P < 0.05) and FAT/CD36 (+16%, P < 0.05) in the heart, and only plasmalemmal FAT/CD36 in muscle (+90%, P < 0.05). Therefore, known changes in the rates of fatty acid oxidation in heart and muscle early in life appear to be accompanied by a concurrent upregulation in the rates of fatty acid transport and the expression of FAT/CD36 in heart and muscle, as well as an increase in plasmalemmal FAT/CD36 and FABPpm in the heart, and only plasmalemmal FAT/CD36 in soleus muscle. We speculate that the rapid upregulation of fatty acid transport rates in heart and muscle are needed to support the increased rates of fatty oxidation that have been previously observed in these tissues. PMID- 16718360 TI - The activities of antioxidant enzymes and the level of malondialdehyde in cerebellum of rats subjected to methotrexate: protective effect of caffeic acid phenethyl ester. AB - Methotrexate (MTX), a folic acid antagonist, is widely used as a cytotoxic chemotherapeutic agent. MTX-associated neurotoxicity is an important clinical problem. The aim of this study was to investigate the role of caffeic acid phenethyl ester (CAPE) on cerebellar oxidative stress induced by MTX in rats. A total of 19 adult male rats were divided into three experimental groups as follows: MTX group (MTX treated), MTX+CAPE group (MTX+CAPE treated), and control group. MTX was administered intraperitoneally (i.p.) with a single dose of 20 mg kg(-1) on the second day of experiment. CAPE was administered i.p. with a dose of 10 micromol kg(-1) day(-1) for 7 days. Malondialdehyde (MDA) levels and activities of superoxide dismutase (SOD) and catalase (CAT) were determined in cerebellar tissue of rats. MTX caused to significant increase in MDA levels (an important marker of lipid peroxidation) in the MTX group compared with the controls (p = 0.006). CAPE significantly reduced the MTX induced lipid peroxidation in the MTX+CAPE group compared to the MTX (p = 0.007). The activities of SOD and CAT were significantly increased in the MTX group when compared with the control group (p = 0.0001, p = 0.004, respectively). The increased activities of these enzymes were significantly reduced by CAPE treatment (p = 0.004, p = 0.034, respectively). As a result, CAPE may protect from oxidative damage caused by MTX treatment in rat cerebellum. PMID- 16718361 TI - Hypercholesterolemia enhances tolerance to lethal systemic hypoxia in middle-aged mice: possible role of VEGF downregulation in brain. AB - Hypercholesterolemia (HCL) is commonly associated with impaired vascular relaxation response and augmented vasoconstriction. Interestingly, it was shown that animals with HCL were less vulnerable to seizures and several clinical studies also revealed a better outcome after stroke in the patients with HCL. To this context, the present study was designed to test the hypothesis that HCL would enhance the animals' resistance to severe systemic hypoxia and in turn prolong their survival time under such noxious condition. Four groups of middle aged (mean age: 51.1 +/- 2.8 weeks) male C57BL/6J wild-type mice (C57BL-WT) and low-density lipoprotein receptor knockout mice (LDLR-KO) were included in the study: two groups were exposed to severe normobaric hypoxia (5% F(I)O(2)) and other two groups were used for brain tissue sample collection and Western blot analysis. The survival time under the hypoxic condition was recorded for each animal. Individual blood samples were collected immedtately after the cessation of spontaneous breathing for measuring plasma total cholesterol (TCL) and triglycerides. The results show that the hypoxia survival time was longer in LDLR KO than C57BL-WT (i.e. 3.7 +/- 0.5 versus 2.3 +/- 0.2 min; P < 0.05). A positive correlation was found between TCL and the survival time (r (2) = 0.43; P < 0.05). Furthermore, a significant downregulation of vascular endothelial growth factor (VEGF) was observed in the brain tissue of LDLR-KO, as compared with C57BL-WT (n, = 3/group; P < 0.05), whereas expression of heme oxygenase 1 was similar in these two groups. We conclude that HCL enhances resistance to lethal systemic hypoxia (i.e. 61% increase in survival time) in middle-aged mice. This paradoxical protective effect of HCL was associated with a concomitant downregulation of cerebral VEGF expression, which could potentially blunt the hypoxia-triggered and VEGF-mediated pathophysiological events leading to death. PMID- 16718362 TI - Short term training attenuates opening of the mitochondrial permeability transition pore without affecting myocardial function following ischemia reperfusion. AB - Opening of the mitochondrial permeability transition pore (PTP) is known to occur during reperfusion of the ischemic heart and to cause dysfunction and injury. The purpose of the present study was to determine whether short-term training (treadmill dunning for 5 days, 30 m.min(-1), 0%) in male Sprague Dawley rats reduces the occurrence of PTP opening in the ischemic-reperfused heart. Hearts from control (C) and trained (T) rats perfused in the Langendorff mode were submitted to ischemia-reperfusion (I-R: 30 and 40 min respectively). In situ PTP opening was quantified using the mitochondrial 2-deoxy [(3)H]glucose ([(3)H]DOG) entrapment method. Following I-R, the recovery of intact mitochondria upon isolation was significantly greater in T vs C hearts (11.7 +/- 0.5 vs 9.1 +/- 0.4 mU citrate synthase.g(-1) wet ventricles, p < or = 0.01). Training also reduced the entrapment of mitochondrial [(3)H]DOG normalized for the loss of intact mitochondria (14.4 +/- 1.4 vs 9.6 +/- 0.8 [(3)H]DOG ratio units, p < or = 0.01). However, under the experimental conditions used the recovery of contractile function, coronary flow and release of LDH in the coronary effluent were similar in both experimental groups. Taken together, these results suggest that short term training can confer mitochondrial protection and reduce PTP opening. PMID- 16718363 TI - Pluripotency potential of human adipose-derived stem cells marked with exogenous green fluorescent protein. AB - Musculoskeletal tissues regeneration requires rapid expansion of seeding cells both in vitro and in vivo while maintaining their multilineage differentiation ability. Human adipose-derived stem cells (ASCs) are considered to contain multipotent mesenchymal stem cells. Monolayer cultures of human ASCs were isolated from human lipoaspirates and passaged 3 times and then infected with replication-incompetent adenoviral vectors carrying green fluorescent protein (Ad/GFP) genes. Then, Ad/GFP infected human ASCs were transferred to osteogenic, chondrogenic, adipogenic, and myogenic medium. The morphological characterization of induced cells was observed using phase-contrast microscopy and fluorescence microscopy. The expression of marker proteins or genes was measured by immunocytochemical and RT-PCR analysis. Osteopontin (OPN), and osteocalcin (OCN) were positive in osteogenic lineages, aggrecan and SOX9 were positive in chondrogenic ones, peroxisome proliferator-activated receptor (PPAR-gamma2) and lipoprotein lipase (LPL) were positive in adipogenic ones, and myogenin and myod1 was positive in myogenic ones. At the same time, the results of fluorescence microscopic imaging proved that the high level of GFP expression during ASCs differentiation maintained stable nearly 2 months. So the exogenous GFP and multilineage potential of human ASCs had no severe influences on each other. Since the human ASCs can be easily obtained and abundant, it is proposed that they may be promising candidate cells for further studies on tissue engineering. Imaging with expression of GFP facilitates the research on ASCs physiological behavior and application in tissue engineering during differentiation both in vitro and in vivo. PMID- 16718364 TI - ZBTB34, a novel human BTB/POZ zinc finger protein, is a potential transcriptional repressor. AB - BTB/POZ protein family plays a key role in many biological processes both in Drosophila and vertebrates through regulating the transcriptional activities of some downstream genes. Here, we obtained a novel member of human BTB/POZ protein family, named as ZBTB34 (Zinc finger and BTB domain containing 34), which encodes 504 amino acid residues with a BTB/POZ domain at its N-terminus that is similar to the same domain of other known transcription regulators. RT-PCR analysis indicated that ZBTB34 was expressed ubiquitously in most adult human tissues, and whilst immunofluorescence assays showed that ZBTB34 was mainly localized to nucleus. Interestingly, the reporter assay in mammalian cells suggested that ZBTB34 might function as a transcriptional repressor. This present work as the first report about the functional exploration of the novel ZBTB34 gene would be contributed to profound understanding of the transcriptional regulation via BTB/POZ protein family. PMID- 16718365 TI - Effect of ascorbic acid supplementation on certain oxidative stress parameters in the post reperfusion patients of myocardial infarction. AB - Reperfusion injury causes oxidative stress thereby resulting in an imbalance between oxidant-antioxidant systems. In the present communication, the effect of ascorbic acid supplementation has been studied on certain oxidant and antioxidant parameters in the blood of the patients with myocardial infarction before and after thrombolysis. In patients after thrombolysis, the activity of antioxidant enzyme, superoxide dismutase, in the blood was found to be significantly reduced where as the activity of the oxidant enzyme, xanthine oxidase, was found to be significantly increased. Malondialdehyde levels, the index of free radical mediated damage, was also found to be significantly elevated in thrombolysed patients compared to the patients before thrombolysis. Supplementation of vitamin C to the post reperfusion patients restored these parameters back to normal or near normal levels. PMID- 16718366 TI - Pesticides induced oxidative stress in thymocytes. AB - The role of oxidative stress in immune cell toxicity caused by the pesticides lindane, malathion and permethrin was investigated in thymic cells from C57BL/6 mice. Thymocytes treated with any of these pesticides (concentrations ranging between 50-150 microM) were found to generate both superoxide ((*)O(2) (-)) and H(2)O(2). The production of (*)O(2) (-) was detected with hydroethidine-ethidium bromide assay. H(2)O(2) production was monitored with a flow cytometric fluorescent (DCFH-DA) assay. All three pesticides stimulated (*)O(2) (-) release after 5 min exposure. Lindane and permethrin, but not malathion, continued to have significant (p < or = 0.05) effects on (*)O(2) (-) generation following 15 min of exposure. The lindane + malathion mixture was found to cause more-than additive increase in (*)O(2) (-) production compared to individual pesticide treatments (at both 5 and 15 min). However, the effect of the lindane + permethrin mixture was not significantly different than individual components of this mixture. The effects of these pesticides on levels of antioxidant enzymes were also investigated, and only mixtures were found to have significant (p < or = 0.05) effects. Thus, lindane + malathion and lindane + permethrin mixtures increased total superoxide dismutase (SOD) specific activity, had no effect on catalase levels and inhibited GSH-peroxidase and GSH-reductase specific activities. Although the results of these studies do not explain the mechanism of action of these pesticides on the generation of (*)O(2) (-) and H(2)O(2), it is worthy of note that mixtures of these chemicals have oxidative responses greater than those of single chemicals. PMID- 16718367 TI - Erythrocyte oxidant/antioxidant status in essential hyperhidrosis. AB - Essential hyperhidrosis is a disorder of excessive, bilateral, and relatively symmetric sweating occurring in the axillae, palms, soles, or craniofacial region without obvious etiology. Nitric oxide may play a physiological part in the production and/or excretion of sweat in skin eccrine glands. Tempol, a SOD mimetic, increases the half-life of NO and results in vasodilatation, hypotension, and reflex activation of sympathetic nervous system. Reactive oxygen species (ROS) may directly activate both central and peripheral sympathetic nervous system activity. We assessed the levels of malondialdehyde (MDA), the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) of red blood cells in patients with essential hyperhidrosis (n = 31) compared to age-and sex-matched healthy controls (n = 28). Erythrocyte activities of SOD and level of MDA were detected significantly higher (p = 0.020, p = 0.004 and respectively) and activities of CAT and GSH-Px were significantly lower (p = 0.0001, p = 0.0001 respectively) in patients than controls. Our results support the hypothesis that oxidative damage resulting from increased ROS production along with insufficient capacity of antioxidant mechanisms may be involved in pathogenesis of EH. PMID- 16718368 TI - A sphingolipid rich lipid fraction isolated from attenuated Leishmania donovani promastigote induces apoptosis in mouse and human melanoma cells in vitro. AB - Lipids, especially sphingolipids, are emerging as inducer of apoptosis in a wide range of immortal cells, potentiating their therapeutic application in cancer. In the present study, a sphingolipid rich lipid fraction (denoted here as ALL), isolated from an attenuated strain of Leishmania donovani promastigote, was tested for its tumoricidal activity taking melanoma, the dreaded form of skin cancer cells, as model. ALL was found to induce chromatin condensation, internucleosomal DNA fragmentation and phosphatidylserine externalization with enhanced cell population in sub-G1 region in both mouse and human melanoma systems, namely B16F10 and A375 respectively. These are the hallmarks of cells undergoing apoptosis. Further analysis demonstrated that ALL treated melanoma cells showed significant increase in ROS generation, mitochondrial membrane potential depolarization, release of cytochrome c, and caspase-3 activation, which are the events closely involved in apoptosis. These findings indicate that one or more bioactive sphingolipid(s)/ceramide(s) present in ALL could be the causative agent(s) for the induction of apoptosis in melanoma cells. Further studies are thus necessary to identify these specific bioactive sphingolipid(s)/ceramide(s) and to establish their mechanism of action, in order to explore their use as anticancer agents. PMID- 16718369 TI - (-)Schisandrin B is more potent than its enantiomer in enhancing cellular glutathione and heat shock protein production as well as protecting against oxidant injury in H9c2 cardiomyocytes. AB - Effects of schisandrin B enantiomers ((+)Sch B and (-)Sch B) treatment on cellular reduced glutathione (GSH) level and heat shock protein (Hsp)25/70 production were investigated in H9c2 cardiomyocytes. (+)Sch B and (-)Sch B at 6.25 muM produced a time-dependent and biphasic change in cellular GSH level and Hsp25/70 production, with the stimulatory effect of (-)Sch B being more potent. The GSH- and Hsp-enhancing effects were accompanied by a parallel cytoprotection against xanthine oxidase/xanthine-induced toxicity, with the biphasic time course of (+)Sch B- or (-)Sch B-induced protection being superimposed with that of the increase in GSH level but not Hsp25/70 production. The results indicate that ( )Sch B produces more potent enhancing effects on cellular GSH and Hsp production as well as protection against oxidative injury than (+)Sch B in cardiomyocytes. PMID- 16718370 TI - Regulation of connexin-43-mediated growth inhibition by a phosphorylatable amino acid is independent of gap junction-forming ability. AB - The ability of the gap junction phosphoprotein connexin-43 (Cx43) to inhibit DNA synthesis in primary cardiomyocytes is regulated by serine (S) 262, a protein kinase C phosphorylation site that also affects metabolic coupling. We have now examined if the S262-regulated growth suppression is operating in transformed cells and if so whether it depends on gap junction channel forming ability. Serine 262 became phosphorylated in response to protein kinase C stimulation in HEK293 cells transiently expressing either Cx43 or the non-channel-forming carboxy-terminal tail of Cx43 (Cx43CT). Expression of either wild type Cx43 or Cx43CT inhibited DNA synthesis, as did their mutated versions simulating lack of phosphorylation by carrying an S262-to-alanine substitution. The ability to inhibit DNA synthesis was eliminated when expressing mutated versions of either Cx43 or Cx43CT simulating constitutive phosphorylation by carrying an S262-to aspartate substitution. We conclude that S262 phosphorylation cancels growth inhibition by Cx43 independently of channel-forming ability. PMID- 16718371 TI - Physiological basis for effect of physical conditioning on chronic ethanol induced hypertension in a rat model. AB - The study aim was to investigate the interaction of physical conditioning and chronic ethanol ingestion on blood pressure (BP), heart rate (HR), nitric oxide (NO) and oxidants/antioxidants balance in the plasma of rats. Male Fisher rats were divided into four groups of seven animals each and treated as follows: (1) Control (5% sucrose, orally) daily for 12 weeks; (2) ethanol (4 g kg(-1), orally) daily for 12 weeks; (3) exercise training on treadmill plus sucrose daily for 12 weeks and (4) exercise training on treadmill followed by ethanol (4 g kg(-1), orally) daily for 12 weeks. The body weight, BP and HR were recorded every week. The animals were sacrificed under ether anesthesia after 12 weeks, blood collected in heparinzed vials, plasma isolated and analyzed. The results show that exercise training significantly lowered the weight gain 6-12 weeks in ethanol treated rats compared to ethanol alone or control rats. The mean arterial BP was significantly elevated 6-12 weeks after ethanol ingestion without significant alterations in HR. Exercise training lowered the BP close to the normal control values in ethanol fed rats. Ethanol significantly decreased the plasma NO levels, reduced to oxidized glutathione ratio (GSH/GSSG) and antioxidant enzymes-superoxide dismutase (CuZn-SOD, and Mn-SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) activities while plasma NADPH oxidase activity and malondialdehyde (MDA) levels were significantly elevated compared to control. Exercise training significantly restored the depletion of plasma NO levels, GSH/GSSG ratio, and antioxidant enzyme activities and normalized the MDA levels and NADPH oxidase activity in the plasma of ethanol treated rats. The study concluded that physical conditioning attenuates the chronic ethanol-induced hypertension by augmenting the NO bioavailability and reducing the oxidative stress response in the plasma of rats. PMID- 16718372 TI - SREBP activation by antipsychotic- and antidepressant-drugs in cultured human liver cells: relevance for metabolic side-effects? AB - Drug-induced weight gain is a major problem in the treatment of psychiatric disorders, especially with some antipsychotic- and antidepressant drugs. We have recently demonstrated that antipsychotic- and antidepressant drugs activate the SREBP (sterol regulatory element-binding proteins) transcription factors in human and rat glial cells, with subsequent up-regulation of downstream genes involved in cholesterol- and fatty acid biosynthesis. Since stimulation of cellular lipogenesis in the liver could be of relevance for the metabolic side effects of these drugs, we have now investigated the effects of antidepressants, antipsychotic- and mood-stabilizing drugs on cell cultures of human liver cells. For several of the drugs being strongly associated with weight gain (clozapine, imipramine, and amitriptyline), we observed a very pronounced activation of SREBP. Ziprasidone and buproprion, however, which are not associated with weight gain, did hardly stimulate the SREBP system. For haloperidol, olanzapine and mirtazapine, the correspondence between metabolic side effects and SREBP stimulation in liver cells was less obvious. The mood-stabilizers did not increase SREBP activation. The results indicate a relationship between drug induced activation of SREBP in cultured human liver cells and weight gain side effects of antidepressant and antipsychotic drugs. PMID- 16718373 TI - Associations of protein 4.2 with band 3 and ankyrin. AB - Protein-protein and protein-lipid interactions are thought to play the vital role in maintenance and deformation of red blood cell (RBC) membrane. Protein 4.2, a 76-KDa peripheral protein, binds to the cytoplasmic domain of band 3 (CDB3) and also interacts with ankyrin in RBCs. In order to explore the characteristics of protein 4.2-CDB3-ankyrin interactions, three protein 4.2-derived recombinant proteins encompassing amino acid residues 31-200, 1-300, and 187-260 respectively were expressed in Escherichia coli. Their interactions with CDB3 and ankyrin were investigated by using Far-Western blot and pull-down assay. The results showed that the CDB3-binding site of protein 4.2 is located in the region of residues 200-211 and the ankyrin-binding site is located in the region of residues 187-200 of protein 4.2. Our findings also suggested that the ankyrin D34 domain can interact directly with protein 4.2. The proper tertiary structures of these protein 4.2 fragments are essential for protein 4.2-ankyrin interaction. Meanwhile, ankyrin can enhance the interaction between protein 4.2 and CDB3. PMID- 16718374 TI - Differential alterations in metabolic pattern of the spliceosomal UsnRNAs during pre-malignant lung lesions induced by benzo(a)pyrene: modulation by tea polyphenols. AB - The differential alterations of the spliceosomal UsnRNAs (U1, U2, U4, U5, and U6) were reported to be associated with cellular proliferation and development. The attempt was made in this study to analyze the metabolic pattern of the spliceosomal UsnRNAs during the development of pre-malignant lung lesions induced in experimental mice model system by benzo(a)pyrene (BP) and also to see how tea polyphenols, epigallocatechin gallate (EGCG) and epicatechin gallate (ECG), modulate the metabolism of these UsnRNAs during the lung carcinogenesis. No significant changes in the level of the UsnRNAs were seen in the inflammatory lung lesions at 9th week due to treatment of BP. However, there was significant increase in the level of U1 ( approximately 2.5 fold) and U5 ( approximately 47%) in the hyperplastic lung lesions at 17th week. But in the mild dysplastic lung lesions at 26th week, the level of UsnRNAs did not change significantly. Whereas, in the dysplastic lung lesions at 36th week there was significant increase in the level of the U2 ( approximately 2 fold), U4 ( approximately 2.5 fold) and U5 ( approximately 2 fold). Due to the EGCG and ECG treatment the lung lesions at 9th week appeared normal and in the 17th, 26th, and 36th week it appeared as hyperplasia. The level of the UsnRNAs was significantly low in the lung lesions at 9th week (only U2 and U4 by EGCG), at 17th week (only U1 by EGCG/ECG), at 26th week (U1 by ECG; U2, U4 and U5 by EGCG/ECG) and at 36th week (U1 by ECG, U2 and U4 by EGCG/ECG). Whereas, there was significant increase in the level of U5 (by EGCG/ECG) and U6 (by EGCG only) in the lung lesions at 36th and 26th week respectively. This indicates that the metabolism of the spliceosomal UsnRNAs differentially altered during the development of pre-malignant lung lesions by BP as well as during the modulation of the lung lesions by the tea polyphenols. PMID- 16718375 TI - Long-term effect of Trigonella foenum graecum and its combination with sodium orthovanadate in preventing histopathological and biochemical abnormalities in diabetic rat ocular tissues. AB - Trigonella foenum graecum seed powder (TSP) and Sodium Orthovanadate (SOV) have been shown to demonstrate antidiabetic effects by stabilizing glucose homeostasis and carbohydrate metabolism in experimental type-1 diabetes. However their efficacy in controlling histopathological and biochemical abnormalities in ocular tissues associated with diabetic retinopathy is not known. The purpose of this study was to investigate the comparative efficacy of individual as well as combination therapy of TSP and SOV in 8 weeks diabetic rat lens and retina. Retinas and lenses were taken from control, alloxan-induced diabetic rats and diabetic rats treated separately with insulin, 5%TSP, SOV (0.6 mg/ml) and a combined dose of SOV (0.2 mg/ml) and 5%TSP for 60 days. Control and each experimental group had six rats. Alterations in the activities of enzymes HK (hexokinase), AR (aldose reductase), SDH (sorbitol dehydrogenase), G-6-PD (glucose-6-phosphate dehydrogenase), GPx (glutathione peroxidase), GR (glutathione reductase) and levels of metabolites like sorbitol, fructose, glucose, MDA (malondialdehyde) and GSH (reduced glutathione) were measured in the cytosolic fraction of lenses besides measuring blood glucose levels and glycosylated haemoglobin. Histopathological abnormalities were studied in the lens using photomicrography and retina using transmission electron microscopy. Blood glucose, glycosylated haemoglobin levels and polyol pathway enzymes AR and SDH increased significantly causing accumulation of sorbitol and fructose in the diabetic lens and treatment with SOV and TSP significantly (p < 0.05) decreased these to control levels. Similarly, SOV and TSP treatments modulated the activities of HK, G-6-PD, GPx and GR in the rat lens to control values. Ultrastructure of the diabetic retina revealed disintegration of the inner nuclear layer cells with reduction in rough endoplasmic reticulum and swelling of mitochondria in the bipolar cells; and these histopathological events were effectively restored to control state by SOV and TSP treatments. In this study SOV and TSP effectively controlled ocular histopathological and biochemical abnormalities associated with experimental type-1 diabetes, and a combination regimen of low dose of SOV with TSP demonstrated the most significant effect. In conclusion, the potential of SOV and TSP alone or in low dose combination may be considered as promising approaches for the prevention of diabetic retinopathy and other ocular disorders. PMID- 16718376 TI - Chronic heat-shock treatment driven differentiation induces apoptosis in Leishmania donovani. AB - The present study investigates the role of apoptosis in the regulation of cell numbers of Leishmania donovani during the in vitro differentiation of promastigote stage to amastigote stage in axenic conditions. We report that apoptosis is induced in Leishmania donovani due to chronic heat-shock treatment of 37 ( degrees )C that also mediates the differentiation of promastigotes to amastigotes. This is characterized by the fragmentation of DNA, blebbing in the parasite cell membrane, nuclear condensation, formation of preapoptotic bodies and involvement of Ca(++) in the apoptotic process. The flowcytometric analysis shows an early and steep rise in percentage apoptotic nuclei till 48-hour stage of differentiation and then a gradual decline, suggesting synergistic action of Ca(++) ATPase and probably Hsp70. Hsp70 might be rescuing cells from apoptosis in the death signaling pathway. Incubation of the culture with Ca(++) chelator EGTA (1 mM) brings down the percentage of apoptotic nuclei considerably showing thereby that calcium is needed for the process of cell death here that occurs by apoptosis. The survival of the infective individuals appears to be decided by the parasite in the early stages of its differentiation. Our studies show the potential of the physiological temperature of 37 ( degrees )C in inducing apoptosis in Leishmania donovani and the therapeutic use it can be put to. PMID- 16718378 TI - Activity and expression of ecto-5'-nucleotidase/CD73 are increased by thyroid hormones in vascular smooth muscle cells. AB - Extracellular nucleotides ATP, ADP, AMP and adenosine are well known signaling molecules of the cardiovascular system that are involved in several physiological processes: cell proliferation, platelet aggregation, inflammatory processes and vascular tonus. The levels of these molecules are controlled by ecto-NTPDases and ecto-5'-nucleotidase/CD73 (ecto-5'-NT/CD73) actions, which are responsible for the complete ATP degradation to adenosine. The thyroid hormones, thyroxine (T(4)) and triiodothyronine (T(3)), play important roles in the vascular system promoting vasodilatation. Here we investigated the influence of thyroid hormones on the enzyme cascade that catalyzes the interconversion of purine nucleotides in vascular smooth muscle cells (VSMC). Exposure of VSMCs to 50nM T(3) or T(4) did not change ATP and ADP hydrolysis significantly. However, the same treatment caused an increase of 75% in AMP hydrolysis, which was time-dependent but dose independent. Moreover, T(3) treatment significantly increased ecto-5'-NT/CD73 mRNA expression, which suggests a genomic effect of this hormone upon ecto-5' NT/CD73. In addition to the importance of the ecto-5'-NT in cell proliferation and differentiation, its overexpression could result in higher extracellular levels of adenosine, an important local vasodilatator molecule. PMID- 16718379 TI - Iron uptake and release by macrophages is sensitive to propranolol. AB - In this study we have tested the effects of d-propranolol (D-Pro) on the iron uptake, iron release and oxidative response of iron-loaded cells in a cellular model of iron-overload using isolated rat peritoneal macrophages incubated with iron-dextran (Fe-D). Pretreatment of macrophages with D-Pro (5-200 microM) prior to Fe-D exposure decreased the cellular iron content and partially prevented iron release from latex-activated macrophages. Release of reactive oxygen species from activated cells was detected by dichlorodihydrofluorescein (DCDHF, 5 microM) oxidation. We found that loading cells with Fe-D increased their response to latex, which was prevented by the lysosomotropic antioxidant agent D-Pro (10 microM). PMID- 16718377 TI - NFAT3 is required for EGF-induced COX-2 transcription, but neither iNOS transcription nor cell transformation in Cl 41 cells. AB - Epidermal growth factor (EGF) has been reported to act as a tumor promoter in several tissues, such as skin, in association with the induction of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS). However, molecular mechanisms involved in these regulations are not well defined. This study addressed a potential role of nuclear factor of activated T cells 3 (NFAT3) in EGF-induced COX-2 and iNOS transcription and cell transformation in mouse epidermal Cl 41 cells. We found that EGF markedly induced anchorage-independent growth (cell transformation) of Cl 41 cells, as well as COX-2 (> 6-fold) and iNOS (> 5-fold) promoter-dependent transcription. The EGF-induced COX-2 transcription was blocked by knockdown of NFAT3 with NFAT3 siRNA, whereas the transcription of iNOS and cell transformation induced by EGF were not affected. Although our recent studies supported that NFAT3 plays an essential role in chemical carcinogen benzo[a]pyrene-7,8-diol-9,10-epoxide (B[a]PDE)-induced cell transformation, the data presented here demonstrated that NFAT3 is required for EGF-induced COX-2 transcription, but neither iNOS transcription nor cell transformation, indicating that the role of NFAT3 in regulating cell transformation is carcinogen-specific. PMID- 16718381 TI - Analysis of loss of heterozygosity and immunohistochemistry in BRCA1 gene in sporadic breast cancers. AB - BRCA1 is a tumour suppressor gene (TSG), which predisposes cancer to both breast and ovary. The primary objective of the present study is to ascertain the involvement of BRCA1 gene in the pathogenesis of sporadic breast cancer women in Chennai (South India) by analysing its protein expression by immunohistochemistry (IHC) and loss of heterozygosity (LOH) for confirmation of the involvement of TSG in the study population. We found down regulation of BRCA1 protein (54%) in IHC and it was correlated with the clinicopathological parameters of the patients. We found near significant correlation (P < 0.063) between BRCA1 protein expression and clinicopathological parameters. We found 30% LOH in our study and it was also correlated with the clinicopathological parameters. No correlation was found between LOH and clinicopathological parameters. Though we found no correlation, the results revealed in this study support the involvement of BRCA1 TSG in the pathogenesis of sporadic breast cancer women in Chennai (South India). PMID- 16718383 TI - Biofeedback and cognitive coping in the treatment of pediatric habit cough. AB - Habit cough is a persistent "barking" cough that does not have a medical basis. The current study evaluated a biofeedback approach using skin temperature feedback with a family focus in the treatment of an 11-year-old girl diagnosed with habit cough. Treatment consisted of six, one hour sessions with the family for part of the session and then individually with the girl. Individual treatment involved skin-temperature biofeedback to teach relaxation during the coughing episodes. The girl was cough free at the end of the sixth session and remained cough free at the end of a one and two year follow-ups. The use of a single case baseline design demonstrated the reduction of coughing and increase in extracurricular activity. It is, noteworthy that the girl demonstrated a significant ability to increase skin temperature during treatment and when asked to try to control her cough. PMID- 16718382 TI - Oxidant stress and damage in post-ischemic mouse hearts: effects of adenosine. AB - Despite the general understanding that ischemia-reperfusion (I/R) promotes oxidant stress, specific contributions of oxidant stress or damage to myocardial I/R injury remain poorly defined. Moreover, whether endogenous 'cardioprotectants' such as adenosine act via limiting this oxidant injury is unclear. Herein we characterized effects of 20 min ischemia and 45 min reperfusion on cardiovascular function, oxidative stress and damage in isolated perfused mouse hearts (with glucose or pyruvate as substrate), and examined whether 10 microM adenosine modified these processes. In glucose-perfused hearts post-ischemic contractile function was markedly impaired (< 50% of pre-ischemia), cell damage assessed by lactate dehydrogenase (LDH) release was increased (12 +/- 2 IU/g vs. 0.2 +/- 0.1 IU/g in normoxic hearts), endothelial-dependent dilation in response to ADP was impaired while endothelial-independent dilation in response to nitroprusside was unaltered. Myocardial oxidative stress increased significantly, based on decreased glutathione redox status ([GSSG]/[GSG + GSSH] = 7.8 +/- 0.3% vs. 1.3 +/- 0.1% in normoxic hearts). Tissue cholesterol, native cholesteryl esters (CE) and the lipid-soluble antioxidant alpha-tocopherol (alpha TOH, the most biologically active form of vitamin E) were unaffected by I/R, whereas markers of primary lipid peroxidation (CE-derived lipid hydroperoxides and hydroxides; CE-O(O)H) increased significantly (14 +/- 2 vs. 2 +/- 1 pmol/mg in normoxic hearts). Myocardial alpha -tocopherylquinone (alpha-TQ; an oxidation product of alpha -TOH) also increased (10.3 +/- 1.0 vs. 1.7 +/- 0.2 pmol/mg in normoxic hearts). Adenosine treatment improved functional recovery and vascular function, and limited LDH efflux. These effects were associated with an anti oxidant effect of adenosine, as judged by inhibition of I/R-mediated changes in glutathione redox status (by 60%), alpha-TQ (80%) and CE-O(O)H (100%). Provision of 10 mM pyruvate as sole substrate (to by-pass glycolysis) modestly reduced I/R injury and changes in glutathione redox status and alpha-TQ, but not CE-O(O)H. Adenosine exerted further protection and anti-oxidant actions in these hearts. Functional recoveries and LDH efflux correlated inversely with oxidative stress and alpha -TQ (but not CE-O(O)H) levels. Collectively, our data reveal selective oxidative events in post-ischemic murine hearts, which are effectively limited by adenosine (independent of substrate). Correlation of post-ischemic cardiovascular outcomes with specific oxidative events (glutathione redox state, alpha-TQ) supports an important anti-oxidant component to adenosinergic protection. PMID- 16718380 TI - IL-6 signaling via the STAT3/SOCS3 pathway: functional analysis of the conserved STAT3 N-domain. AB - The conserved N-domain of the STAT proteins has been implicated in several activities crucial to cytokine signaling including receptor recruitment and STAT activation, cooperative DNA binding and STAT-dependent gene expression. We evaluated the role of the STAT3 N-domain in the IL-6 signal transduction pathway leading to Socs3 gene expression, an essential mechanism that controls the quality and magnitude of IL-6-dependent transcriptional responses. Based on the model for STAT N-domain function in cooperative gene expression and the presence of tandem STAT binding motifs in the murine Socs3 promoter, we anticipated that stabilizing interactions between adjacent STAT3 dimers via N-domain sequences might be essential for Socs3 gene expression. This was underscored by the tight conservation in the location and sequence of the tandem STAT binding sites between the murine and human Socs3 promoters. Using reconstitution into Stat3-/- mouse embryonic fibroblasts (Stat3-/- MEFs), we find that a STAT3 N-domain deletion mutant (Delta 133STAT3) is activated by tyrosine phosphorylation in response to IL-6 and then undergoes dephosphorylation with kinetics similar to full-length STAT3. These results highlight important differences compared to other STATs where the N-domain has been shown to mediate activation (STAT4) or dephosphorylation (STAT1). STAT3 binds predominantly to a single STAT consensus site in the Socs3 promoter, despite the presence of an adjacent STAT motif. Significantly, Delta 133STAT3 stimulates expression of the endogenous Socs3 gene in Stat3-/- MEFs upon IL-6 treatment with an activity similar to reconstituted STAT3, demonstrating that the N-domain is dispensable for Socs3 gene expression. We propose that the Socs3 gene in its chromosomal context is activated by the IL 6/STAT3 pathway independent of STAT3 N-domain sequences. PMID- 16718397 TI - Including pre-AIDS mortality in back-calculation model to estimate HIV prevalence in France, 2000. AB - Since the advent of highly active antiretroviral therapy (HAART) the lengthening of AIDS incubation time has led to a decrease of AIDS incidence and mortality, and to the increase of the proportion of pre-AIDS mortality. The objective was to develop an extension of the back-calculation model by including pre-AIDS mortality and to estimate HIV prevalence in France. Our previous back-calculation model was modified to take into account the probability of survival for HIV infected individual using the relative risk to die at different period for an HIV infected person versus the general population (psi). AIDS cases reported to InVS (Institut de Veille Sanitaire) until March 2003 were back-calculated to estimate HIV incidence until December 2000. AIDS deaths occurring until December 2000 were used to obtain HIV prevalence in 2000. Plausible intervals were calculated taking into account uncertainties on AIDS incubation time. Taking into account pre-AIDS mortality increased the goodness-of-fit of the model to the data. The relative risk, psi, was estimated as 3 for homo-bisexual men, haemophiliacs and transfused cases, 10 for intravenous drug users, and 4 for heterosexual cases, with no difference over period. HIV prevalence in 2000 was estimated as 88,200 (with a plausible interval of 52,300-168,000), versus 78,200 when mortality was not considered. Pre-AIDS mortality estimates show the amount of this mortality during the era of HAART but also evidenced its existence before HAART. Taking into account pre-AIDS mortality of HIV-infected person in the back-calculation model increased the estimated HIV prevalence. PMID- 16718398 TI - High prevalence of vitamin D deficiency, secondary hyperparathyroidism and generalized bone pain in Turkish immigrants in Germany: identification of risk factors. AB - INTRODUCTION: The aim of the study was to determine the prevalence of vitamin D deficiency, secondary hyperparathyroidism (sHPT), generalized bone pain and predictors of vitamin D deficiency in a cohort of 994 healthy adult urban residents (589 males, 405 females; age range: 16-69 years) consisting of 101 Germans, 327 Turkish residents of Turkey and 566 Turkish immigrants living in Germany. METHODS: The mean (+/- standard deviation) for 25-hydroxyvitamin D [25(OH)D] and biointact parathyroid hormone (BioPTH) for the German men and women was 68.4 nmol/l and 26.7 pg/ml, respectively. Turkish residents of Turkey had a mean 25(OH)D and BioPTH of 40.6 nmol/l and 27.5 pg/ml, respectively, whereas Turkish residents of Germany had a 25(OH)D of 38.1 nmol/l and a BioPTH of 35.6 pg/ml. RESULTS: Vitamin D insufficiency was common among Turkish nationals independent of whether they lived in Turkey or Germany; 75% had 25(OH)D levels of <50 nmol/l. Turkish females had a higher prevalence of 25(OH)D deficiency (<25 nmol/l) than Turkish males: 30 and 19% of Turkish females living in Germany and Turkey were severely vitamin D deficient compared to 8% and 6% of Turkish males living in Germany and Turkey, respectively. With respect to BioPTH levels, 31% of Turkish females and 21% of Turkish males had elevated BioPTH levels in contrast to only 15% of females and 4% of males living in Turkey. Unconditional logistic regression analysis identified the most important predictors for low 25(OH)D levels as sex, body mass index, lack of sun exposure and living at a higher latitude. Additionally, wearing a scarf and number of children were found to be an independent risk factor for vitamin D deficiency in Turkish women living in Turkey and Germany. A strong correlation between low 25(OH)D levels and higher rates and longer duration of generalized bone and/or muscle aches and pains (often diagnosed as fibromyalgia) was observed. CONCLUSION: Secondary hyperparathyroidism and vitamin D deficiency was found to be common among Turkish immigrants living in Germany, especially in veiled women. Therefore, the monitoring of vitamin D status--i.e. 25(OH)D and PTH--in Turkish immigrants is warranted and once a deficiency is identified, it should be appropriately treated. PMID- 16718399 TI - The effect of a low-carbohydrate diet on bone turnover. AB - INTRODUCTION: Low-carbohydrate diets have become popular as weight loss techniques. These diets are high in protein, saturated fats, and omega-6 fatty acids. They also lead to a ketogenic state. These factors could lead to increased bone turnover. This study was designed to see whether a low-carbohydrate diet would lead to increased bone turnover in humans. METHODS: Thirty patients (15 study subjects and 15 controls) were recruited for this 3-month study. The 15 patients on the diet were instructed to consume less than 20 g of carbohydrates per day for the 1st month and then less than 40 g per day for months 2 and 3. Control subjects had no restrictions on their diet. The primary end point was urinary N-telopeptide (UNTx) at 3 months. Secondary end points included UNTx at 1 month, bone-specific alkaline phosphatase (BSAP) at 1 month, bone turnover ratio (BSAP/UNTx) at 1 month, and weight loss. RESULTS: The mean UNTx in the study subjects increased by 1.6 [95% confidence interval (CI) +/-22.8] compared with an increase of 1.9 (95% CI +/-17.6) in the controls at 3 months (p=0.86). The mean UNTx decreased by 2.2 (95% CI +/-27.2) and 3.1 (95% CI +/-17.6) at 1 month in the dieters and controls, respectively (p=0.36). The mean BSAP decreased by 0.53 (95% CI +/-2.96) in the dieters and increased by 0.34 (95% CI +/-2.92) in the controls at 1 month (p=0.27). The bone turnover ratio increased by 0.08 (95% CI +/-0.81) in the dieters and by 0.05 (95% CI +/- 0.27) in the controls at 1 month (p=0.78). The dieters lost 6.39 kg versus 1.05 kg for the controls at 3 months (p=0.0008). CONCLUSIONS: Although the patients on the low-carbohydrate diet did lose significantly more weight than the controls did, the diet did not increase bone turnover markers compared with controls at any time point. Further, there was no significant change in the bone turnover ratio compared with controls. PMID- 16718400 TI - Laryngeal branching pattern of the inferior laryngeal nerve, before entering the larynx. AB - The objective of the present work was to investigate the laryngeal branching pattern of the inferior laryngeal nerve (ILN) in detail before the branches entered the larynx. In 49 specimens 96 sides were examined for this project, including 27 males and 22 female cadavers. In 7 sides (7.3%, 5 on the left and 2 on the right) the ILN passed as a single trunk into the larynx below the inferior constrictor muscle. In 82 sides (85.4%, 40 on the left and 42 on the right) two laryngeal branches originated from the nerve. The nerve divided either just before entering the larynx (58.3%, 24 on the left and 32 on the right) or 15-32 mm below the inferior constrictor muscle (27.1%, 16 on the left and 10 on the right). In 69 sides (71.8%), the anterior and/or posterior laryngeal branches subdivided into one or two sub-branches. The anterior (observed in 49 sides, 51%) and posterior laryngeal branches supplied all intrinsic laryngeal muscles except the cricothyroid muscle and the mucosa below the vocal cords, respectively, while their sub-branches reached the cricopharyngeal part of the inferior constrictor muscle, esophagus and/or the thyroid gland. In 7 sides (7.3%, 4 on the left and 3 on the right) the nerve divided into three laryngeal branches. In conclusion, the branching pattern of the nerve may be important pitfalls of the thyroidal and laryngeal surgery. Owing to this the surgeon should keep in mind the risk of extra laryngeal division of the nerve and not confuse laryngeal and extra laryngeal branches. PMID- 16718402 TI - Investigation of interaction between two neutralizing monoclonal antibodies and SARS virus using biosensor based on imaging ellipsometry. AB - Two neutralizing human scFv, b1 and h12 were identified initially using ELISA,employing highly purified virus as the coating antigen. The biosensor technique based on imaging ellipsometry was employed directly to detect two neutralizing monoclonal antibodies and serial serum samples from 10 SARS patients and 12 volunteers who had not SARS. Further, the kinetic process of interaction between the antibodies and SARS-CoV was studied using the real-time function of the biosensor. The biosensor is consistent with ELISA that the antibody h12 showed a higher affinity in encountering the virus than antibody b1. The affinity of antibody b1 and antibody h12 was 9.5 x 10(6) M(-1) and 1.36 x 10(7) M(- 1), respectively. As a label free method, the biosensor based on imaging ellipsometry proved to be a more competent mechanism for measuring serum samples from SARS patients and the affinity between these antibodies and the SARS coronavirus. PMID- 16718401 TI - Anatomy of right recurrent nerve node: why does early metastasis of esophageal cancer occur in it? AB - Early, distant and/or skip metastasis of squamous-cell thoracic esophageal cancer frequently occurs in the right recurrent nerve node (recR). However, the specific lymphatic route without a nodal relay, such as the submucosal ascending route, was not known for the recR afferent. Using 20 donated cadavers, macroscopic, and histological observations were performed on the recR and its surrounding lymphatics, especially afferent routes from the esophagus to the recR. Most afferent vessels of the recR originated from the right paratracheal node. However, the recR often (12/20) received a major submucosal lymphatic drainage route ascending along the thoracic esophagus. The submucosal vessel came out of the esophagus and ran in a longitudinal connective tissue mass along the right tracheo-esophageal groove. A direct drainage route was often (13/20) seen from the recR to the venous system. Moreover, because of the specific histology, collaterals seemed to be present around the recR. In the regional nodes of the intrathoracic esophagus, the recR histology was characterized by the high proportion of lymphocyte accumulating areas or the cortex. From the midthoracic level, metastatic cancer cells seemed to reach the recR via esophageal submucosal vessels in the early stage. Large lymphocyte accumulating areas of the recR suggested higher filtration capacity than other distal nodes. However, the collateral of the recR and its direct drainage to the venous system suggested that the recR involvement often corresponds to a systemic disease. PMID- 16718403 TI - Design of a novel MEMS platform for the biaxial stimulation of living cells. AB - Micromechanical systems are increasingly being used as tools in biological applications, since their characteristic dimensions permit to operate at the same length scale of the structures under investigation. Here, we present a methodology for the design, fabrication and operation of a tool for the assessment of mechanical properties of single cells. In particular, we describe a microsystems platform to study bio-mechanical response of single living cells to in-plane biaxial stretching. The proposed device employs a new linkage design in order to obtain the displacement of the quadrants of a sliced circular plate in mutually-orthogonal directions using just one linear actuator. With this linkage geometry, the whole device has only one degree of freedom. This results in a very predictable and reliable mechanical behaviour, thereby allowing use a simple and easily available control electronics. Results of this study have relevance for the design of a powerful yet simple BioMEMS platform for the characterization of living cells as in-plane bi-axial loading simulated the conditions experienced by cells in vivo more realistically than a uniaxial stretching. PMID- 16718404 TI - 3D tissue culture substrates produced by microthermoforming of pre-processed polymer films. AB - We describe a new technology based on thermoforming as a microfabrication process. It significantly enhances the tailoring of polymers for three dimensional tissue engineering purposes since for the first time highly resolved surface and bulk modifications prior to a microstructuring process can be realised. In contrast to typical micro moulding techniques, the melting phase is avoided and thus allows the forming of pre-processed polymer films. The polymer is formed in a thermoelastic state without loss of material coherence. Therefore, previously generated modifications can be preserved. To prove the feasibility of our newly developed technique, so called SMART = Substrate Modification And Replication by Thermoforming, polymer films treated by various polymer modification methods, like UV-based patterned films, and films modified by the bombardment with energetic heavy ions, were post-processed by microthermoforming. The preservation of locally applied specific surface and bulk features was demonstrated e.g. by the selective adhesion of cells to patterned microcavity walls. PMID- 16718406 TI - Integrated polymerase chain reaction chips utilizing digital microfluidics. AB - This study reports an integrated microfluidic chip for polymerase chain reaction (PCR) applications utilizing digital microfluidic chip (DMC) technology. Several crucial procedures including sample transportation, mixing, and DNA amplification were performed on the integrated chip using electro-wetting-on-dielectric (EWOD) effect. An innovative concept of hydrophobic/hydrophilic structure has been successfully demonstrated to integrate the DMC chip with the on-chip PCR device. Sample droplets were generated, transported and mixed by the EWOD-actuation. Then the mixture droplets were transported to a PCR chamber by utilizing the hydrophilic/hydrophobic interface to generate required surface tension gradient. A micro temperature sensor and two micro heaters inside the PCR chamber along with a controller were used to form a micro temperature control module, which could perform precise PCR thermal cycling for DNA amplification. In order to demonstrate the performance of the integrated DMC/PCR chips, a detection gene for Dengue II virus was successfully amplified and detected. The new integrated DMC/PCR chips only required an operation voltage of 12V(RMS) at a frequency of 3 KHz for digital microfluidic actuation and 9V(DC) for thermal cycling. When compared to its large-scale counterparts for DNA amplification, the developed system consumed less sample and reagent and could reduce the detection time. The developed chips successfully demonstrated the feasibility of Lab-On-a-Chip (LOC) by utilizing EWOD-based digital microfluidics. PMID- 16718407 TI - Dielectrophoresis tweezers for single cell manipulation. AB - Positioning single cells is of utmost importance in areas of biomedical research as diverse as in vitro fertilization, cell-cell interaction, cell adhesion, embryology, microbiology, stem cell research, and single cell transfection. Here we describe dielectrophoretic tweezers, a sharp glass tip with electrodes on either side, capable of trapping single cells with electric fields. Mounted on a micromanipulator, dielectrophoresis tweezers can position a single cell in three dimensions, holding the cell against fluid flow of hundreds of microns per second with more than 10 pN of force. We model the electric field produced by the tweezers and the field produced by coaxial microelectrodes. We show that cells are trapped without harm while they divide in the trap. In addition, dielectrophoretic tweezers offer the possibility for trapping, electroporating, and microinjecting a single cell with one probe. PMID- 16718405 TI - Biological application of multi-component nanowires in hybrid devices powered by F1-ATPase motors. AB - In this paper, construction of hybrid device by integrating nanowires with F(1) ATPase motors is described. The nickel nanowires and multi-segment nanowires, including gold and nickel, were fabricated by electrochemical deposition in nanoporous templates. The nickel nanowires functionalized by biotinylated peptide can be assembled directly onto F(1)-ATPase motors to act as the propellers. If the multi-component nanowires, including gold and nickel, were selectively functionalized by the thiol group modified ssDNA and the synthetic peptide, respectively, the biotinylated F(1)-ATPase motors can be attached to the biotinylated peptide on nickel segment of the nanowires. Then, the multi component nanowires can also be used as the propellers, and one may observe the rotations of the multi-component nanowires driven by F(1)-ATPase motors. Therefore, introduction of multiple segments along the length of a nanowire can lead to a variety of multiple chemical functionalities, which can be selectively bound to cells and special biomolecules. This method provides an insight for the construction of other hybrid devices with its controlling arrangement of different biomolecule on designed nanometer scale structures. PMID- 16718424 TI - Variations of patient doses in interventional examinations at different angiographic units. AB - PURPOSE: We analyzed doses for various angiographic procedures using different X ray systems in order to assess dose variations. METHODS: Dose-area product (DAP), skin doses from thermoluminescent dosimeters and air kerma measurements of 308 patients (239 diagnostic and 69 interventional) were assessed for five different angiographic units. All fluoroscopic and radiographic exposure parameters were recorded online for single and multiprojection studies. Radiation outputs of each X-ray system were also measured for all the modes of exposure using standard protocols for such measurements. RESULTS: In general, the complexity of the angiographic procedure was found to be the most important reason for high radiation doses. Skill of the radiologist, management of the exposure parameters and calibration of the system are the other factors to be considered. Lateral cerebral interventional studies carry the highest risk for deterministic effects on the lens of the eye. Effective doses were calculated from DAP measurements and maximum fatal cancer risk factors were found for carotid studies. CONCLUSIONS: Interventional radiologists should measure patient doses for their examinations. If there is a lack of necessary instrumentation for this purpose, then published dose reports should be used in order to predict the dose levels from some of the exposure parameters. Patient dose information should include not only the measured quantity but also the measured radiation output of the X-ray unit and exposure parameters used during radiographic and fluoroscopic exposures. PMID- 16718423 TI - A new device for vascular embolization: report on case of two pulmonary arteriovenous fistulas embolization using the amplatzer vascular plug. AB - A pulmonary arteriovenous fistula (PAVF) is a rare vascular malformation commonly treated by embolization with coils or balloons to prevent the risk of several serious complications such as cerebral embolism and brain abscess. A 32-year-old female with two PAVFs and neurological ischemic manifestations has been successfully treated by transcatheter embolization of both fistulas using a new device (Amplatzer Vascular Plug). This self-expanding cylindrical nitinol mesh cage with high radial strength allows a chance of relocation until properly positioned. It is preferred to coils or balloons because a large caliber of feeding artery implied high risk of uncontrollable distal embolization. There appear to be no reports in the literature concerning use of this device, which could represent a useful innovative tool in embolotherapies, especially in large vascular areas. PMID- 16718425 TI - Nontraumatic rupture of lumbar artery causing an intravertebral body pseudoaneurysm: treatment by transcatheter embolization. AB - We report a case of nontraumatic rupture of the lumbar artery that led to a pseudoaneurysm in the vertebral body and massive retroperitoneal to retropleural hematoma. The pseudoaneurysm was successfully treated by endovascular embolization. To our knowledge, idiopathic rupture of the lumbar artery has been reported in a limited number of cases and pseudoaneurysm formation in the vertebral body has not been presented in the literature. The etiology of hemorrhage has been discussed based on CT, MRI, and three-dimensional rotational angiography. PMID- 16718426 TI - Deployment of stents in two separate colorectal strictures in a patient with extracolonic pelvic metastases. PMID- 16718427 TI - Mid-term results after endovascular stent-grafting of descending aortic aneurysms in high-risk patients. AB - PURPOSE: To analyze our experience with endovascular stent-grafting of descending aortic aneurysms in high-risk patients. METHODS: Nineteen patients underwent endovascular stent-graft repair of descending aortic aneurysms using the Talent Stent Graft System (Medtronic). All patients were considered high-risk for open surgical repair due to their age, requirement for emergency surgery, and comorbidities. Computed tomography and/or MR tomography were performed at 3, 6 and 12 months postoperatively and thereafter every 12 months. RESULTS: Secondary technical success was 100%. Thirty-day mortality was 5%. Incidence of postoperative stroke and paraplegia were 5% each. One patient required a second stent-graft due to a type I endoleak during the same hospital stay (primary technical success 95%). All patients have been followed for a median of 20 months. No migration, wire fractures or endoleak appeared during follow-up. CONCLUSION: Endovascular stent-grafting had a low 30-day mortality and morbidity in high-risk patients. One patient developed an aortoesophageal fistula 40 days after stent implantation. Stent-graft repair is a valuable supplement to surgical therapy in high-risk patients. PMID- 16718428 TI - Endovascular treatment of supra-aortic extracranial stenoses in patients with vertebrobasilar insufficiency symptoms. AB - PURPOSE: Stenoses and thromboses of vessels feeding the vertebrobasilar territory can evoke serious disturbances including ischemic stroke. We present our experience of endovascular interventions for patients with signs of vertebrobasilar insufficiency (VBI) resulted from subclavian, vertebral and brachiocephalic arterial stenoses. METHODS: Twenty-one patients (10 men) aged from 35 to 84 years (mean 64.3 years) with symptoms compatible with VBI underwent balloon angioplasty and stenting of subclavian (SA), innominate (IA) and vertebral (VA) arteries. Procedures were done by radiologists experienced in systematic stenting of the lesions. VBI was manifested by persistent signs in 15 patients, and by transitory ischemic attacks in the posterior circulatory territory in 4 (19%). Two patients (10%) experienced ischemic strokes (in the vertebrobasilar circulation in both cases). In 3 patients (14%) VBI was accompanied by upper limb vascular insufficiency symptoms. All cases were resistant to medical treatment. A neurologist assessed complaints, initial VBI signs and their alteration after intervention in all patients. Outcomes were measured with the 5-point scale suggested by Malek et al.: (1) excellent result (asymptomatic, no neurologic deficits and no symptoms of vertebrobasilar ischemia); (2) good (no neurologic deficits, at most one transient episode of vertebrobasilar ischemia over a period of 3 months after treatment); (3) fair (minimal neurologic deficit and at most one transient episode per month of vertebrobasilar ischemia); (4) poor (no improvement compared with neurologic status before treatment and/or persistent symptoms of vertebrobasilar ischemia); (5) death (regardless of cause). Endovascular treatment was performed for SA stenosis in 15 patients, for SA occlusion in 2, for IA stenosis in 2, and for VA stenosis in 2. There were 15 cases of atherosclerosis, 2 of aortoarteritis, 4 of proximal SA kinking. SA and IA stenoses ranged from 60% to 100% (mean 74.5%), VA stenoses were 90% in both cases. RESULTS: Initial technical success was achieved in 96% of cases. There were no postprocedural complications or deaths. During 6 36 months (mean 21.3 months) of follow-up all patients showed improvement in VBI symptoms or upper limb ischemia. Within 36 months after the procedure outcomes were estimated as excellent and good in 13 patients (76%) and poor in 2 (12%), the last being attributed to atherosclerosis progression in other vascular areas. Restenosis in the stented area has developed in 1 patient (6%). CONCLUSIONS: Balloon angioplasty and stenting of extracranial vertebrobasilar arterial stenoses appeared to be effective in endovascular treatment of medically resistant VBI. Further investigations are required to clarify the role of subclavian artery kinking in VBI development and indications for various methods of its correction. PMID- 16718432 TI - Saccadic instabilities in albinism without nystagmus. AB - Albinism effects a surprising manipulation of the visual pathway in which some of the normally uncrossed axons of the temporal retina instead cross at the chiasm. An expected consequence of this misrouting is that subjects with albinism will have difficulty in specifying the targets of saccades. Usually albinos have nystagmus so the stability of their saccadic eye movements is not readily accessible, but some albinos do not have nystagmus. In these subjects it was found that they had frequent saccadic intrusions, the sizes of which were correlated with velocities of steady drifts in fixations (r = 0.802, P < 0.05). An explanation for the correlation between the amplitudes of the intrusions and the velocities of the drifts is that it is due to a common failure in the development of a saccadic system which is responsible for converting a given retinal displacement into a matching eye movement, with the extent of the failure reflecting the severity of the misrouting. PMID- 16718433 TI - Upregulation of persistent and ramp sodium current in dorsal horn neurons after spinal cord injury. AB - Traumatic spinal cord injury (SCI) results not only in motor impairment, but also in chronic central neuropathic pain, which often is refractory to conventional treatment approaches. Upregulated expression of sodium channel Nav1.3 has been observed within the spinal dorsal horn neurons after SCI, and appears to contribute to neuronal hyperresponsiveness and pain-related behaviors. In this study we characterized the changes in sodium current properties within dorsal horn neurons after contusive SCI. Four weeks after adult male Sprague-Dawley rats underwent T9 spinal cord contusion injury, when behavioral nociceptive thresholds were decreased to both mechanical and thermal stimuli, whole-cell patch-clamp recordings were performed on acutely dissociated lumbar dorsal horn neurons. The cells demonstrated characteristic fast-activating and fast-inactivating sodium currents. SCI led to a shift of the steady-state activation and inactivation of the sodium current towards more depolarized potentials. The shifted steady-state inactivation shows similarities to that obtained from axotomized dorsal root ganglions, which were shown to upregulate Nav1.3. Small slow depolarizations below action potential threshold produced ramp currents, which were markedly enhanced by SCI (from 182 +/- 41 to 338 +/- 55 pA). The density of the noninactivating persistent sodium current was also significantly enhanced in neurons from SCI animals (from 17.4 +/- 3.2 to 27.7 +/- 4.4 pA/pF at 50-70 ms of depolarization). The increased persistent sodium current and ramp current, which are consistent with upregulation of Nav1.3 within dorsal horn neurons, suggest a basis for the hyperresponsiveness of these neurons following SCI. PMID- 16718434 TI - Phylogeography of the genus Cardiandra based on genetic variation in cpDNA sequences. AB - We investigated the phylogenetic relationships within the genus Cardiandra based on plastid DNA sequences. The phylogenetic tree showed that Cardiandra populations from the Ryukyu Islands (Japan) and Taiwan were monophyletic (Ryukyu Taiwan clade), whereas taxa from China and mainland Japan were sisters to this clade. The divergence time between the Ryukyu-Taiwan clade and the other species was estimated to be 0.082 MYA, i.e., the late Pleistocene. The infrageneric and/or infraspecific differentiation of Cardiandra is estimated to have depended largely on allopatric differentiation caused by the presence or division of the past landbridge of the Ryukyu Islands, which connected mainland Japan to the Asian Continent during the Quaternary. PMID- 16718445 TI - Perivascular nerve damage in the cerebral circulation following traumatic brain injury. AB - Traumatic brain injury (TBI) causes cerebral vascular dysfunction. Most have assumed that it was the result of endothelial and/or smooth muscle alteration. No consideration, however, has been given to the possibility that the forces of injury may also damage the perivascular nerve network, thereby contributing to the observed abnormalities. To test this premise, we subjected rats to impact acceleration. At 6 h, 24 h and 7 days post-TBI, cerebral basal arteries were removed and processed with antibody targeting protein gene product 9.5 (PGP-9.5), with parallel assessments of 5-hydroxytryptamine (5-HT) accumulation in the perivascular nerves. Additionally, Fluoro-Jade was also used as a marker of axonal degeneration. The perivascular nerve network revealed no abnormality in sham animals. However, by 6 h post injury, Fluoro-Jade reactivity appeared in the perivascular regions, with the number of fibers increasing with time. By 24 h post injury, a significant reduction in the perivascular 5-HT accumulation occurred, together with a reduction in PGP-9.5 fiber staining. At 7 days, a recovery of the PGP-9.5 immunoreactivity occurred, however, it did not reach a control-like distribution. These studies suggest that neurogenic damage occurs following TBI and may be a contributor to some of the associated vascular abnormalities. PMID- 16718449 TI - Tempol prevents harmful effects of remote ischemia reperfusion injury on healing of experimental colonic anastomoses. AB - BACKGROUND AND AIMS: Tempol (4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl) is a water-soluble analogue of the spin label TEMPO. As an antioxidative agent, it is a member of nitroxides, which detoxifies superoxide and possibly other toxic radicals in vivo. In this study, we aimed to investigate whether tempol prevents harmful systemic effects of superior mesenteric ischemia-reperfusion on left colonic anastomosis in rats. MATERIALS AND METHODS: Anastomosis of the left colon was performed in 30 rats that were divided into three groups each having ten animals: sham-operated control (group I), 60 min of intestinal ischemia reperfusion by superior mesenteric artery occlusion (group II), and tempol treated group (30 mg/kg before and after the ischemia-reperfusion (group III). On postoperative day 5, all animals were killed and anastomotic bursting pressures were measured in vivo. Tissue samples were obtained for further investigation of anastomotic hydroxyproline content, perianastomotic malondialdehyde, and glutathione levels. RESULTS: There was a statistically significant increase in the quantity of myeloperoxidase activity and malondialdehyde levels in group II, along with a decrease in glutathione levels, anastomotic hydroxyproline content, and bursting pressure values when compared to controls. However, all of the investigated parameters were normalized in tempol-treated animals (group III). CONCLUSION: We conclude that tempol significantly prevents harmful systemic effects of reperfusion injury on colonic anastomoses in a rat model of superior mesenteric artery occlusion. PMID- 16718450 TI - The clinical value of a negative multi-detector computed tomographic angiography in patients suspected of coronary artery disease: A meta-analysis. AB - The aim of this meta-analysis was to calculate the sensitivity of contrast enhanced multi-detector computed tomography (MDCT) compared with coronary angiography (CAG) in incident patients suspected of coronary artery disease (CAD). We searched PubMed, Embase, bibliographies of original papers and reviews to identify original papers including > or =20 patients. Two independent reviewers selected papers and judged eligible papers on quality. Heterogeneity was assessed and homogeneous subgroups were pooled. Of the 15 included studies, ten provided moderately homogeneous patient-based analyses with absolute diagnostic numbers (n = 630 patients). Pooled sensitivity was 89% (95% confidence interval: 85-92%). Scanners with 16 detectors (n = 4) had higher sensitivities (pooled sensitivity: 91%) than four-detector scanners (n = 6; pooling not possible due to heterogeneity). Seven studies reported sensitivity for a proximal stenosis, but different definitions were used making pooling impossible; sensitivities ranged from 75 to 100%. The sensitivity of four- and 16-detector MDCT is not sufficient to rule out any stenosis in patients suspected of CAD. No conclusions can be drawn with respect to the sensitivity for clinically relevant or proximal stenoses. PMID- 16718451 TI - The radiologic evaluation of tinnitus. AB - Tinnitus ("ringing in the ears") is a prevalent symptom in the general population, and often brings patients to medical attention. Many causes of tinnitus are evident radiographically. The most frequently-encountered causes of tinnitus are discussed, and imaging recommendations are provided. PMID- 16718452 TI - The impact of motion artifacts on the reproducibility of repeated coronary artery calcium measurements. AB - The purpose of this study is, using a 16-section multidetector-row helical computed tomography (MDCT) scanner with retrospective reconstruction, to compare variability in repeated coronary calcium scoring and qualitative scores of the motion artifacts. One hundred forty-four patients underwent two subsequent scans using MDCT. According to Agatston and volume algorithms, the coronary calcium scores during mid-diastole (the center corresponding to 70% of the R-R cycle) were calculated and the inter-scan variability was obtained. Motion artifacts from coronary artery calcium were subjectively evaluated and classified using a 5 point scale: 1, excellent; no motion artifacts; 2, fine, minor motion artifacts; 3, moderate, mild motion artifacts; 4, bad, severe motion artifacts; 5, poor, doubling or discontinuity. Each reading was done by vessels (left main, left descending, left circumflex and right coronary arteries) and the motion artifact score (mean of the scales) was determined per patient. The variability in the low (1.2+/-0.2) and high (2.4+/-0.6) motion artifact score groups was 7+/-6 (median, 6)% and 19+/-15 (16)% on the Agatston score (P<0.01) and 7+/-7 (6)% and 16+/-13 (14)% on the volume score (P<0.01), respectively. In conclusion, motion has a significant impact on the reproducibility of coronary calcium scoring. PMID- 16718453 TI - A narrative account of the impact of positive thinking on discussions about death and dying. AB - GOALS OF WORK: The purpose of this study was to explore the experience of autologous stem cell transplant recipients (ASCT) and those who care for them. MATERIALS AND METHODS: This was a qualitative prospective, longitudinal study. Ten patients who were about to have ASCT and nine carers were recruited to the study. Interviews were to be conducted at regular intervals six times over 2 years. The narratives of two widowed carers were analysed using Grounded Theory and read for themes on positive thinking and death. MAIN RESULTS: Positive thinking has a range of meanings, and its use can have a range of consequences. It can either be a useful coping strategy or can interfere with important conversations and planning about the end of life, and subsequently add to the distress of a grieving partner. CONCLUSIONS: It is important for patients, their partners and their health professionals to be able to discuss potential adverse consequences of illness, including death, without being hindered by the obligation to be positive or optimistic. PMID- 16718456 TI - The dying patient: new Israeli legislation. PMID- 16718461 TI - [Coma. The prognostic value of evoked potentials in patients after traumatic brain injury]. AB - BACKGROUND: The prognosis after traumatic coma is often unclear. We investigated the prognostic value of somatosensory (SSEP) and early acoustic (EAEP) evoked potentials on comatose patients in the intensive care unit regarding long-term outcome. Different evaluation systems were investigated. METHODS: This was a retrospective analysis of 100 patients. SSEP and EAEP were examined at different times and analysed according to the Riffel score. Combinations of the different types of potentials were evaluated regarding possible improvement of outcome prediction. RESULTS: The positive predictive value of at least one missing peak V of the EAEP regarding a fatal prognosis was 83%. The negative predictive value of the EAEP was 96%. A good outcome (GOS 4+5) could be predicted by bilateral normal SSEP and EAEP with a positive predictive value of 98%. CONCLUSIONS: Early evaluation of SSEP and EAEP allows reliable prognostic predictions regarding a later outcome in patients with severe traumatic brain injury and should therefore be used more often for intensive care patients. PMID- 16718460 TI - Cerebral perfusion computerized tomography: influence of reference vessels, regions of interest and interobserver variability. AB - INTRODUCTION: There are still no standardized guidelines for perfusion computerized tomography (PCT) analysis. METHODS: A total of 61 PCT studies were analyzed using either the anterior cerebral artery (ACA) or the middle cerebral artery (MCA) as the arterial reference, and the superior sagittal sinus (SSS) or the vein of Galen (VG) as the venous reference. The sizes of regions of interest (ROI) were investigated comparing PCT results obtained using a hemispheric ROI combined with vascular pixel elimination with those obtained using five smaller ROIs located over the cortex and basal ganglia. In addition, interobserver variations were explored using a standardized protocol. RESULTS: MCA-based measurements of cerebral blood flow (CBF) and blood volume (CBV) were in accordance with those obtained with the ACA except in 16 patients with ischemic stroke, in whom CBF was overestimated by the ipsilateral MCA. Venous maximal intensity was significantly lower with the VG when compared with the SSS, resulting in overestimation of CBF and CBV. However, in 13.3% of patients the VG ROI yielded higher maximal intensities than the SSS ROI. There was no difference in PCT results between hemispheric ROI and averaged separate ROI when vascular pixel elimination was used. Finally, interobserver variations were as high as 11% for CBF and 12% for CBV. CONCLUSION: The present results suggest that pathological rather than anatomical considerations should dictate the choice of the arterial ROI. For venous ROI, although SSS seems to be adequate in most instances, deep cerebral veins may occasionally generate higher maximal intensities and should therefore be selected. Importantly, significant user dependency should be taken into account. PMID- 16718462 TI - IL-1beta-induced pro-apoptotic signalling is facilitated by NCAM/FGF receptor signalling and inhibited by the C3d ligand in the INS-1E rat beta cell line. AB - AIMS/HYPOTHESIS: IL-1beta released from immune cells induces beta cell pro apoptotic signalling via mitogen-activated protein kinases (MAPKs) and nuclear factor-kappaB (NF-kappaB). In neurons, the neural cell adhesion molecule (NCAM) signals to several elements involved in IL-1beta-induced pro-apoptotic signalling in beta cells. Pancreatic beta cells express NCAM, but its biological effects in these cells are unclear. The aim of this study was to investigate whether there is cross-talk between NCAM signalling and cytokine-induced pro-apoptotic signalling. MATERIALS AND METHODS: Western blotting was used to investigate levels of NCAM and inducible nitric oxide synthase, phosphorylation of Src and MAPKs, and cleavage of caspase-3. MAPK activity was investigated with an in vitro kinase assay. Apoptosis was detected by cleaved caspase-3 and a Cell Death Detection ELISA(plus) assay. NCAM-induced fibroblast growth factor receptor (FGFR) activation was investigated in NCAM(-/-) Trex293 cells where FGFR phosphorylation was measured by Western blotting after NCAM transfection. RESULTS: Pre-exposure of INS-1E cells to the FGFR-inhibitor SU5402, but not to the Src-inhibitor PP2, dose-dependently inhibited IL-1beta-mediated MAPK activity. A synthetic peptide, C3d, reported to bind NCAM, did not activate MAPK or Akt as reported in neurons but inhibited IL-1beta-induced MAPK activity, thereby mimicking the effect of SU5402. Furthermore, C3d inhibited NCAM-induced FGFR phosphorylation and apoptosis induced by IL-1beta plus IFN-gamma, but did not affect IL-1beta-induced NF-kappaB signalling. CONCLUSIONS/INTERPRETATION: We suggest that NCAM signalling through FGFR is required for efficient IL-1beta pro apoptotic signalling by facilitating IL-1beta-induced MAPK activation downstream of the NF-kappaB-MAPK branching point. Further, these data identify a novel function of C3d as an inhibitor of NCAM-induced FGFR activity and of IL-1beta induced MAPK activation in beta cells. PMID- 16718464 TI - Understanding of diabetes prevention studies: questionnaire survey of professionals in diabetes care. AB - AIMS/HYPOTHESIS: Diabetes prevention studies have reported reductions of diabetes risk by up to 60%. Since the underlying metabolic changes are small, the clinical significance of this effect may be overestimated. The present survey explores the extent to which different formats of presenting study results may influence diabetes healthcare professionals' perceptions of the importance of intervention effects on diabetes risk. SUBJECTS, MATERIALS AND METHODS: Participants of three European diabetes conferences (160 nurse educators, 112 physicians, 27 other professionals) were presented with a questionnaire that included nine items, in which results from three diabetes prevention studies were presented in different ways. RESULTS: Participation rate was 96%. Effects were interpreted as important or very important by 92% (255/276) when results were presented as proportions of subjects with diabetes (14% intervention group, 29% control group), by 87% (248/285) when results were communicated as a risk reduction of 57%, by 39% (110/284) when the corresponding fasting plasma glucose values were presented (mean difference 0.3 mmol/l), and by 18% (52/283) when glycosylated haemoglobin values were used (6.0 vs 6.1%). Corresponding results of the three diabetes prevention studies were rated as being of identical importance by only 23, 13 and 16% of participants, respectively. CONCLUSIONS AND INTERPRETATION: Healthcare professionals rate the benefit of preventive interventions substantially higher when changes in diabetes risk are communicated rather than related glycaemic parameters. Transformation of continuous metabolic data into diagnostic categories may impair understanding of study effects. PMID- 16718463 TI - Signalling mechanisms linking hepatic glucose and lipid metabolism. AB - Fatty liver and hepatic triglyceride accumulation are strongly associated with obesity, insulin resistance and type 2 diabetes, and are subject to nutritional influences. Hepatic regulation of glucose and lipid homeostasis is influenced by a complex system of hormones, hormonally regulated signalling pathways and transcription factors. Recently, considerable progress has been made in elucidating molecular pathways and potential factors that are affected in insulin resistant states. In this review we discuss some of the key factors that are involved in both the regulation of glucose and lipid metabolism in the liver. Understanding the molecular network that links hepatic lipid accumulation and impaired glucose metabolism may provide targets for dietary or pharmacological interventions. PMID- 16718465 TI - Renal amino acid, fat and glucose metabolism in type 1 diabetic and non-diabetic humans: effects of acute insulin withdrawal. AB - AIMS/HYPOTHESIS: The aim of this study was to test the hypothesis that type 1 diabetes alters renal amino acid, glucose and fatty acid metabolism. MATERIALS AND METHODS: We studied five C-peptide-negative, type 1 diabetic subjects during insulin replacement (glucose 5.6 mmol/l) and insulin deprivation (glucose 15.5 mmol/l) and compared them with six non-diabetic subjects. Leucine, phenylalanine, tyrosine, glucose and palmitate tracers were infused after an overnight fast and samples were obtained from the renal vein, femoral vein and femoral artery. RESULTS: Insulin deprivation significantly increased whole-body fluxes (20-25%) of phenylalanine, tyrosine and leucine, and leucine oxidation (50%). Kidney contributed 5-10% to the whole-body leucine and phenylalanine flux. A net uptake of phenylalanine, conversion of phenylalanine to tyrosine (5 micromol/min) and net release of tyrosine (approximately 5 micromol/min) occurred across the kidney. Whole-body (three-fold) and leg (two-fold) leucine transamination increased but amino acid metabolism in the kidney did not alter with diabetes or insulin deprivation. Insulin deprivation doubled endogenous glucose production, renal glucose production was unaltered by insulin deprivation and diabetes (ranging between 100 and 140 micromol/min). Renal palmitate exchange was unaltered by insulin deprivation. CONCLUSIONS/INTERPRETATION: In conclusion, kidney post-absorptively accounts for 5-10% of whole-body protein turnover, 15 20% of leucine transamination and 10-15% of endogenous glucose production, and actively converts phenylalanine to tyrosine. During insulin deprivation, leg becomes a major site for leucine transamination but insulin deprivation does not affect renal phenylalanine, leucine, palmitate or glucose metabolism. Despite its key metabolic role, insulin deprivation in type 1 diabetic patients does not alter many of these metabolic functions. PMID- 16718467 TI - The economics of inclusion body processing. AB - Many recombinant proteins are often over-expressed in host cells, such as Escherichia coli, and are found as insoluble and inactive protein aggregates known as inclusion bodies (IBs). Recently, a novel process for IB extraction and solubilisation, based on chemical extraction, has been reported. While this method has the potential to radically intensify traditional IB processing, the process economics of the new technique have yet to be reported. This study focuses on the evaluation of process economics for several IB processing schemes based on chemical extraction and/or traditional techniques. Simulations and economic analysis were conducted at various processing conditions using granulocyte macrophage-colony stimulating factor, expressed as IBs in E. coli, as a model protein. In most cases, IB processing schemes based on chemical extraction having a shorter downstream cascade demonstrated a competitive economic edge over the conventional route, validating the new process as an economically more viable alternative for IB processing. PMID- 16718468 TI - Impact of extreme temperatures on daily mortality in Madrid (Spain) among the 45 64 age-group. AB - This paper analyses the relationship between extreme temperatures and mortality among persons aged 45-64 years. Daily mortality in Madrid was analysed by sex and cause, from January 1986 to December 1997. Quantitative analyses were performed using generalised additive models, with other covariables, such as influenza, air pollution and seasonality, included as controls. Our results showed that impact on mortality was limited for temperatures ranging from the 5th to the 95th percentiles, and increased sharply thereafter. During the summer period, the effect of heat was detected solely among males in the target age group, with an attributable risk (AR) of 13.3% for circulatory causes. Similarly, NO(2) concentrations registered the main statistically significant associations in females, with an AR of 15% when circulatory causes were considered. During winter, the impact of cold was exclusively observed among females having an AR of 7.7%. The magnitude of the AR indicates that the impact of extreme temperature is by no means negligible. PMID- 16718469 TI - Paclitaxel nanoparticle inhibits growth of ovarian cancer xenografts and enhances lymphatic targeting. AB - OBJECTIVES: Ovarian cancer has the highest mortality of all the gynecologic cancers. The antitumor agent paclitaxel has been proved to be efficient in the treatment of ovarian cancer. Our study is to develop a polymeric drug delivery system for paclitaxel and determine whether paclitaxel nanoparticle can inhibit growth of ovarian carcinoma xenografts in Fisher344 (F344) rats by intraperitoneal administration. The mechanism of paclitaxel nanoparticles in rats bearing ovarian cancer has been investigated in this study. METHODS: Synthesize paclitaxel loading nanoparticle (PLA) by ultrasonic emulsification; MTT analysis identified cytotoxic activity of paclitaxel nanoparticle in vitro; rat ovarian carcinoma cells were injected into the peritoneal cavity of F344 rats. The antitumor effect of paclitaxel nanoparticle in vivo has been evaluated by measuring tumor weight and ascite volume. At the end of the procedure rats were sacrificed; tumors were excised and processed for PCNA staining, tissue terminal deoxynucleotide transferase-mediated dUTP nick and labeling assay and RT-PCR to evaluate the proliferative and apoptotic changes and cancer transfer-related gene expression induced by PLA. Paclitaxel concentration in plasma, pelvic lymph nodes, liver, and heart were determined by high-performance liquid chromatography. RESULTS: Paclitaxel nanoparticle and PTX (Cremophor) showed equivalent cytotoxic activity in vitro. In rats implanted carcinoma cells, paclitaxel nanoparticles significantly reduced tumor weight and ascites volume, and induced apoptosis of tumor cells. PLA also inhibited cell proliferation and matrix metalloproteinase 9 mRNA expression. The paclitaxel concentration of pelvic lymph nodes in PLA treated animals was 20-fold higher than that of free PTX treated animals at 48 h after intraperitoneal administration. CONCLUSION: The intraperitoneal administration of paclitaxel nanoparticle can significantly inhibit the progression of ovarian carcinoma in peritoneal cavity of female F344 rat. The paclitaxel nanoparticle is safe and lymphatic targeting. PMID- 16718470 TI - Pegylated liposomal doxorubicin in combination with vinorelbine as salvage treatment in pretreated patients with advanced breast cancer: a multicentre phase II study. AB - PURPOSE: To investigate the activity and tolerance of pegylated liposomal doxorubicin in combination with vinorelbine in pretreated patients with metastatic breast cancer. PATIENTS AND TREATMENT: Thirty-six women with metastatic breast cancer were enrolled. The median age was 64 years, 80% of the patients had a performance status of 0-1, 30 (83%) had visceral disease and 83% had received prior taxanes while 50% anthracyclines. Treatment consisted of pegylated liposomal doxorubicin (40 mg/m2 on day 1) and vinorelbine (25 mg/m2 on days 1 and 15) every 4 weeks. RESULTS: In an intention-to-treat analysis 2 (6%) complete and 12 (33%) partial responses were observed (overall response rate 39%; 95% CI: 23-54.8%); 8 (22%) and 14 (39%) patients experienced stable and progressive disease, respectively. The median TTP was 6.5 months and the median survival time 14.2 months. The 1-year survival rate was 54.1%. Grade 3 and 4 neutropenia occurred in 21 (58%) patients, grade 3-4 anemia in four (11%) and grade 4 thrombocytopenia in one (3%). Two (6%) patients developed febrile neutropenia. Non-hematologic toxicity was mild and easily manageable. There was no clinically important cardiac toxicity or treatment-related deaths. CONCLUSIONS: The combination of pegylated liposomal doxorubicin and vinorelbine is an active and well tolerated salvage regimen in patients with metastatic breast cancer which merits further evaluation. PMID- 16718472 TI - Melanocyte differentiation antigen RAB38/NY-MEL-1 induces frequent antibody responses exclusively in melanoma patients. AB - Expression pattern and immunogenicity are critical issues that define tumor antigens as diagnostic markers and potential targets for immunotherapy. The development of SEREX (serological analysis of recombinant expression libraries) has provided substantial progress in the identification of tumor antigens eliciting both cellular and humoral immune responses in cancer patients. By SEREX, we have previously identified RAB38/NY-MEL-1 as a melanocyte differentiation antigen that is highly expressed in normal melanocytes and melanoma tissues but not in other normal tissues or cancer types. In this study, we further demonstrate that RAB38/NY-MEL-1 is strongly immunogenic, leading to spontaneous antibody responses in a significant proportion of melanoma patients. The immune response occurs solely in malignant melanoma patients and was not detected in patients with other diseases, such as vitiligo, affecting melanocytes. Fine analysis of the spontaneous anti-RAB38/NY-MEL-1 antibody response reveals a polyclonal B cell recognition targeting various epitopes, although a dominant immunogenic region was preferentially recognized. Interestingly, our data indicate that this recognition is not rigid in the course of a patient's response, as the dominant epitope changes during the disease evolution. Implications for the understanding of spontaneous humoral immune responses are discussed. PMID- 16718473 TI - Improved perceived health status persists three months after a special sports camp for children with congenital heart disease. AB - INTRODUCTION: Although summer and sports camps for children with congenital heart disease are organized in many countries and regions, empirical data on the effects of such camps is limited. OBJECTIVES: The aim of the present study was to investigate changes in the perceived health status and habitual physical activities in children attending a special sports camp. MATERIALS AND METHODS: In this longitudinal study, 25 children with congenital heart disease who participated in a three-day multi-sports camp were included. The perceived health status was measured using the Child Health Questionnaire-Child Form, CHQ-CF87, completed by the child at the start of the camp (T1), at the end of the camp (T2), and 3 months after the camp concluded (T3). Habitual physical activities were assessed by means of a modified version of the Baecke questionnaire, which was completed by one of the parents at T1 and T3. RESULTS: During the sports camp, we observed significant improvements in the children's perception of their physical functioning, role-physical functioning, general health, role-emotional functioning, self-esteem, mental health, and general behavior. For physical functioning, role-emotional functioning, and family activities, high scores persisted three months after the sports camp concluded. The habitual physical activities (sport and leisure time) of the children remained unchanged. DISCUSSION AND CONCLUSION: In conclusion, we propose that a special sports camp for children with congenital heart disease may improve specific dimensions of subjective health status. Our study confirms a previous report on the benefits of such camps for afflicted children. If these findings can be further corroborated in other settings, participation in sport camps should be advocated as a simple, noninvasive means to promote healthier children. PMID- 16718474 TI - Seasonality in pediatric asthma admissions: the role of climate and environmental factors. AB - BACKGROUND: Population-based data from Taiwan are used to examine seasonality in pediatric asthma admissions (proxy for asthma exacerbations) and associations with air pollutants and climatic factors. Monthly admission rates per 100,000 population, classified into three age groups, 0 approximately 2, 2 approximately 5, and 6 approximately 14 years (calculated from a total of 27,275 hospitalizations during 1998-2001) were subjected to autoregressive integrated moving average (ARIMA) modeling to examine seasonality. Spearman rank correlations were used to examine associations with criterion air pollutants (PM(10), SO(2), CO, O(3), NO(2)) and meteorological factors (ambient temperature, relative humidity, atmospheric pressure, rainfall, and sunshine hours). RESULTS: Both seasonality and associations with air pollutants and climate factors vary by age group. Among under-twos, the rates are lowest in January-February and highest in November, with a trough in June-July. Among preschoolers, the rates are lowest in June-July and highest in November, with two upsurges in August and March. Among school-goers, admission rates are lowest during June-August, with upsurges in March and September. The number of weather and pollutant predictors increases with age. Among under-twos, only two factors, PM(10) and rainfall, significantly predict admissions. For preschoolers, five factors (PM(10), CO, O(3), temperature, and pressure), and for school-goers, all air pollutants except NO(2,) and all climatic factors except rainfall are significant. CONCLUSION: Seasonality in pediatric asthma admissions vary by age in a subtropical island setting. PMID- 16718477 TI - Osteoporotic femoral fracture in a child with propionic acidaemia presenting as non-accidental injury. PMID- 16718476 TI - Overweight and obesity and their associated factors in adolescents in Tehran, Iran, 2004-2005. AB - OVERVIEW: Obesity is a significant health crisis around the world. Of great concern are the data pointing to the recent increase in the prevalence of obesity irregardless of age group and country. Overweight and obesity in adolescence are markers of overweight and obesity in adults, respectively. Very little data are currently available on the prevalence of childhood obesity in Iran, and more research on the risk factors is required before preventive public health programs can be formulated and put into practice. OBJECTIVE: The objective of this study was to quantify the prevalence of overweight and obesity and their associated factors in adolescent children living in Tehran. MATERIALS AND METHODS: During a multistage stratified cluster sampling, 2900 students (1200 males and 1700 females) aged 11-17 years were selected from 20 secondary schools in the school year of 2004-2005. A questionnaire was filled, and weight and height were measured. DISCUSSIONS AND CONCLUSIONS: The body mass index (BMI) was calculated and adjusted for age and sex. Prevalences of overweight and obesity were 17.9 and 7.1%, respectively. BMI increased with age, and it was higher in those who had lower levels of physical activity. Age at menarche was negatively associated with BMI. There was no relationship between macro- and micronutrient intake and overweight and obesity. This study highlights the high prevalence of overweight and obesity in adolescent children in Tehran. PMID- 16718478 TI - Maternal vitamin A deficiency and neonatal microphthalmia: complications of biliopancreatic diversion? AB - Biliopancreatic diversion (BPD) for morbid obesity carries a serious risk of nutritional deficiencies that might impair embryogenesis. Consequently, attention should be given to the potential of risk to the fetus of BPD in women of childbearing age. We present the case of a pregnant woman who had undergone BPD 8 years previously, with documented vitamin A deficiency, who gave birth to a child with bilateral microphthalmia. Infectious and genetic causes of microphthalmia were excluded. A search of the literature revealed that vitamin A deficiency may cause a disruption of ocular development. We conclude that nutritional deficiencies may cause a spectrum of fetal malformations. As the effect of BPD relies on malabsorption, fetal risk should be considered before BPD is offered to morbid obese women of childbearing age. PMID- 16718479 TI - Avian stanniocalcin-2 is expressed in developing striated muscle and joints. AB - The glycoprotein hormone stanniocalcin (STC) has originally been described in the teleost kidney. Since then, STC homologs have been identified in various genomes including human, mouse, rat, Xenopus and zebrafish. In mammals, two STC genes, STC1 and STC2, are known. We cloned a chicken STC homolog to analyze its expression pattern during chick development. Sequence analyses revealed a high sequence similarity of the chicken STC (cSTC) clone to mammalian STC2. Interestingly the expression pattern of cSTC2 largely resembles those of murine STC1: we found expression of cSTC2 in the nephric tubules, in the myocardium, in skeletal muscle cells from the onset of differentiation, and in synovial joint anlagen of the limbs. PMID- 16718480 TI - Research in child and adolescent psychopharmacology: recent accomplishments and new challenges. AB - RATIONALE: Research in pediatric psychopharmacology has expanded considerably in the last 10 years. Still, controversy remains about the effectiveness and safety of commonly used psychotropics and their role in child treatment, thus pointing to the need for more in-depth and targeted investigations. OBJECTIVES: To review recent accomplishments and current limitations of pediatric psychopharmacology, and discuss approaches to further research. METHODS: Selective review of the relevant literature and research in progress. RESULTS: Controlled clinical trials have been conducted in many common psychiatric disorders in children and adolescents, thus providing a basis on which evidence-based treatment guidelines can be constructed. Little innovation has, however, occurred in treatment development and testing. Safety concerns are prominent and have a major influence on clinical practice and drug utilization. CONCLUSIONS: While a research infrastructure has been successfully built for conducting pediatric clinical trials, important aspects such as long-term treatment effects, optimal sequencing and individualization of interventions, and integration of neuroscience findings into innovative, theory-driven treatment development remain to be addressed. PMID- 16718483 TI - Functional roles of the pepper pathogen-induced bZIP transcription factor, CAbZIP1, in enhanced resistance to pathogen infection and environmental stresses. AB - Transcription factors often belong to multigene families and their individual contribution in a particular regulatory network remains difficult to assess. We identify and functionally characterize the pepper bZIP transcription factor CAbZIP1 gene isolated from pepper leaves infected with Xanthomonas campestris pv. vesicatoria. Transient expression analysis of the CAbZIP1-GFP fusion protein in Arabidopsis protoplasts revealed that the CAbZIP1 protein is localized in the nucleus. The N-terminal region of CAbZIP1 fused to the GAL4 DNA-binding domain is required to activate transcription of reporter genes in yeast. The CAbZIP1 transcripts are constitutively expressed in the pepper root and flower, but not in the leaf, stem and fruit. The CAbZIP1 gene is locally or systemically induced in pepper plants infected by either X. campestris pv. vesicatoria or Pseudomonas fluorescens. The CAbZIP1 gene is also induced by abiotic elicitors and environmental stresses. The CAbZIP1 transgenic Arabidopsis exhibits a dwarf phenotype, indicating that CAbZIP1 may be involved in plant development. The CAbZIP1 overexpression in the transgenic Arabidopsis plants confers enhanced resistance to Pseudomonas syringae pv. tomato DC3000, accompanied by expression of the AtPR-4 and AtRD29A. The transgenic plants also exhibit increased drought and salt tolerance during all growth stages. Moreover, the transgenic plants are tolerant to methyl viologen-oxidative stress. Together, these data suggest that the CAbZIP1 transcription factor function as a possible regulator in enhanced disease resistance and environmental stress tolerance. PMID- 16718482 TI - The signal attenuation rat model of obsessive-compulsive disorder: a review. AB - During the last 30 years, there have been many attempts to develop animal models of obsessive-compulsive disorder (OCD), in the hope that they may provide a route for furthering our understanding and treatment of this disorder. The present paper reviews a recently developed rat model of OCD, namely, signal attenuation. Results of pharmacological and lesion studies are presented and evaluated with respect to the pharmacology and pathophysiology of OCD. It is argued that signal attenuation is a rat model of OCD with construct (derived from similarity in the underlying mechanisms), predictive (derived from similarity in response to treatment), and face (derived from phenomenological similarity between "compulsive" behavior in the model and compulsions in OCD patients) validity. PMID- 16718484 TI - Co-expression of the borage Delta 6 desaturase and the Arabidopsis Delta 15 desaturase results in high accumulation of stearidonic acid in the seeds of transgenic soybean. AB - Two relatively rare fatty acids, gamma-linolenic acid (GLA) and stearidonic acid (STA), have attracted much interest due to their nutraceutical and pharmaceutical potential. STA, in particular, has been considered a valuable alternative source for omega-3 fatty acids due to its enhanced conversion efficiency in animals to eicosapentaenoic acid when compared with the more widely consumed omega-3 fatty acid, alpha-linolenic acid (ALA), present in most vegetable oils. Exploiting the wealth of information currently available on in planta oil biosynthesis and coupling this information with the tool of genetic engineering it is now feasible to deliberately perturb fatty acid pools to generate unique oils in commodity crops. In an attempt to maximize the STA content of soybean oil, a borage Delta(6) desaturase and an Arabidopsis Delta(15) desaturase were pyramided by either sexual crossing of transgenic events, re-transformation of a Delta(6) desaturase event with the Delta(15) desaturase or co-transformation of both desaturases. Expression of both desaturases in this study was under the control of the seed-specific soybean beta-conglycinin promoter. Soybean events that carried only the Delta(15 )desaturase possessed a significant elevation of ALA content, while events with both desaturases displayed a relative STA abundance greater than 29%, creating a soybean with omega-3 fatty acids representing over 60% of the fatty acid profile. Analyses of the membrane lipids in a subset of the transgenic events suggest that soybean seeds compensate for enhanced production of polyunsaturated fatty acids by increasing the relative content of palmitic acid in phosphatidylcholine and other phospholipids. PMID- 16718485 TI - EST generation and analyses towards identifying female gametophyte-specific genes in Zea mays L. AB - The embryo sac (female gametophyte) plays an important role in double fertilization. The female gametophyte is composed of four specific cell types: the synergids that attract pollen tubes, the egg cell and central cell which are fusion partners for the two sperm cells, and the antipodal cells whose function is unknown. As a resource for gene discovery and to help identify genes exhibiting cell-specific expression patterns, we constructed cDNA libraries from female gametophytes and from egg cells of maize and sequenced more than 8,500 ESTs. These libraries represent diverse transcripts, potentially corresponding to 3,850 genes (contigs and singletons) from the female gametophyte and 963 genes (contigs and singletons) from the egg cell. In each collection, 16% of the contigs/singletons have no matches in databases and 3-5% encode hypothetical proteins; novel hypothetical proteins (not found within the female gametophyte contigs) were identified among the egg cell contigs. We examined 65 contigs by RT PCR and 19 genes that were potentially female gametophyte-specific were identified. We used in situ hybridization to determine expression specificity for seven genes: one transcript was expressed both in the egg cell and in the central cell, one was expressed in the egg cell and synergids, two were expressed in the central cell, two were expressed in the synergids, and one was expressed in the central cell and the synergids. Four of these encode small, potentially secreted peptides that are dissimilar except for a conserved triple cysteine motif near their C-terminus. These EST resources should prove useful for identifying female gametophyte or cell-specific genes. PMID- 16718490 TI - [The MIS anterolateral approach for THA]. AB - The MIS anterolateral approach for hip replacement surgery uses the intermuscular plane between gluteus medius and tensor fascia latae. The patient is placed in the lateral position. Retractors with soft tissue friendly profile and an operating table with modified leg support are recommended. The technique provides very good exposure of the acetabulum and femur and preserves muscle integrity so that rehabilitation can be rapid. Currently the experience comprises over 1000 primary cases performed using the MIS anterolateral approach since March 2003. The clinical results including pain, blood loss, range of motion, functional score, Harris Hip Score, and radiographic studies are based on the complete examinations of 48 consecutive patients. There are significant differences with improved clinical results compared with patients after standard approaches. After a fundamental learning curve there is no evidence of increased complications. The anterolateral approach is a standard approach suitable for wide application. The experience includes difficult primary cases like dysplasia, rheumatic disease, and ankylosis and even for revision surgery the technique can be routinely used. Restrictions are not known either in patients' characteristics or in indications. PMID- 16718491 TI - Femoral artery occlusion after blunt trauma: diagnosis by multislice CT angiography. PMID- 16718492 TI - A physiologically based pharmacokinetic (PB/PK) model for multiple exposure routes of soman in multiple species. AB - A physiologically based pharmacokinetic (PB/PK) model has been developed in advanced computer simulation language (ACSL) to describe blood and tissue concentration-time profiles of the C(+/-)P(-) stereoisomers of soman after inhalation, subcutaneous and intravenous exposures at low (0.8-1.0 x LD(50)), medium (2-3 x LD(50)) and high (6 x LD(50)) levels of soman challenge in three species (rat, guinea pig, marmoset). Allometric formulae were used to compute the compartment volumes, blood flow rates, tidal volume and respiratory rate based upon total animal weight. Blood/tissue partition coefficients for soman, initial carboxylesterase and acetylcholinesterase levels and the rate constants for interactions between soman and these enzymes were species-dependent and were obtained from in vitro measurements reported in the literature. The model incorporated arterial and venous blood, lung, kidney, liver, richly perfused, poorly perfused and fat tissue compartments as well as subcutaneous and nasal exposure site compartments. First-order absorption from linearly filled soman deposits into metabolizing exposure site compartments was employed to model subcutaneous and inhalation exposures. The model was validated by comparing the predicted and observed values for C(+/-)P(-)-soman in arterial blood at various times following exposure and by regression analysis. Sensitivity analysis was used to determine the effects of perturbations in the model parameters on the time-course of arterial C(-)P(-)-soman concentrations for different exposure routes. In our evaluation of 28 datasets, predicted values were generally within 95% confidence limits of the observed values, and regression coefficients comparing predicted and observed data were greater than 0.85 for 95% of the intravenous and subcutaneous datasets and 25% of the inhalation datasets. We conclude that the model predicts the soman toxicokinetics for doses >or=1 x LD(50) for intravenous and subcutaneous exposures and inhalation exposures of 8 min or less sufficiently well to allow its use in the modeling of bioscavenger protection. PMID- 16718493 TI - RelA1 gene control of Escherichia coli lipid structure and cell performance during glucose limited fed-batch conditions. AB - At increasing glucose limitation, typical for fed-batch cultivation performance, cultivation of Escherichia coli (relA1) results in development of a lipid structure that radically differs from the wild type and is characterised by accumulation of neutral phospholipids and saturated fatty acids. The mutant can, furthermore, not change the level of cardiolipin, which is generally the hallmark of changes to severe glucose limitation. The result suggests an increased negative control in the mutant with respect to the flux to phosphatidyl glycerol and cardolipin as well as to unsaturated fatty acids. Opposite to the wild type, the cardiolipin-depleted membrane is more fragile with respect to sonication and osmotic chock, at severe limitation, and results in extensive foaming during the process. Protein leakage and cell lysis is, however, lower in the mutant most likely due to the increased amounts of saturated fatty acids, which might be a possible strategy to overcome the reduced amounts of membrane-strengthening cardiolipin. The membrane potential of the outer surface is negative, however less negative for the mutant. This was supported by aqueous two-phase extraction experiments which, furthermore indicated a difference in outer surface hydrofobicity. These findings suggest that the relA1 gene has a defined, but ppGpp-independent, role in cells with a slowly decreasing metabolism of glucose to control the membrane morphology. PMID- 16718494 TI - What are the risk factors in the development of osteoarthritis following ACL reconstruction? PMID- 16718495 TI - In vivo stability of total knee arthroplasty using a navigation system. AB - The aim of this study was to evaluate in vivo stability for mediolateral laxity in extension and anteroposterior laxity in 90 degrees of flexion and to correlate these and the range of motion (ROM) in 42 total knee arthroplasties (TKA) performed using a navigation system, with a minimum 1-year follow-up. The following parameters were measured at the final follow-up: mediolateral laxity in extension and anteroposterior laxity in 90 degrees of flexion as determined by stress radiographs and a Telos arthrometer, modified HSS scores (excluding laxity and range of motion) and the range of motion (ROM). The mean modified HSS score was 82% of 82 maximum allowable points, and the mean postoperative ROM was 128.1+/-10.4 degrees . Mean medial laxity was 3.5+/-1.4 degrees , mean lateral laxity was 4.4+/-2.2 degrees and mean anteroposterior laxity was 7.1+/-4.1 mm. We found no significant correlation between mediolateral laxity and postoperative ROM. However, a significant correlation was found between postoperative ROM and anteroposterior laxity. In conclusion, the use of a navigation system in TKA assists the surgeon to achieve good in vivo stability. Short-term clinical results are promising. PMID- 16718496 TI - Identification and characterization of HLA-class-I-restricted T-cell epitopes in the putative tumor-associated antigens P21-activated serin kinase 2 (PAK2) and cyclin-dependent kinase inhibitor 1A (CDKN1A). AB - Multiple myeloma (MM) is one of the most common hematological malignancies. Despite a variety of therapeutical approaches including high-dose cytostatic treatment with subsequent autologous or allogeneic stem cell transplantation, as well as vaccination, cures remain rare exceptions. An important issue for future immunological treatments is the identification and characterization of appropriate tumor-associated antigens. However, the number of tumor-associated antigens in MM is limited. PBK/TOPK and activated serin kinase 2 (PAK2) are novel serin kinases that have recently been identified. PBK/TOPK is overexpressed in Burkitt lymphoma, acute lymphoblastic leukemia, and MM; PAK2 is expressed in malignant lymphatic cells. The cyclin kinase inhibitor 1A (CDKN1A) is overexpressed in MM compared to normal plasma cells. We hereby identified and characterized for the first time HLA-class-I-restricted immunogenic peptides in the amino acid sequences of PAK2 and CDKN1A. Using two independent prediction algorithms, we identified two peptides in PAK2 and three peptides in CDK1NA with high binding to HLA-A2. Using an IFN-gamma ELISPOT assay, we could demonstrate the presence and functional activity of CD8-peptide-specific T cells with all tested peptides. To show HLA-A2-restricted antigen recognition, the specific inhibition of T cell recognition was demonstrated with an anti-HLA-A2-blocking antibody. By analysis of peripheral blood of 34 healthy donors for the presence and functional activity of CD8 T cells specific for these peptides, we could demonstrate that peptide T-cell precursors specifically recognizing at least one of the tested peptides are present in 50-60% of the tested donors and that these T-cell precursors can be expanded in vitro. We conclude that PAK2- and CDKN1A derived peptides can elicit a strong and consistent CD8 T-cell response in an in vitro model. Further investigations will examine the presence and functionality of such T cells in the tumor-bearing host. PMID- 16718497 TI - Relapse of acute promyelocytic leukemia presenting as granulocytic sarcoma in the hip. PMID- 16718498 TI - Thalidomide for POEMS syndrome. PMID- 16718499 TI - The discrete dynamics of monotonically decomposable maps. AB - We extend results of Gouze and Hadeler (in Nonlinear World 1:23-34, 1994) concerning the dynamics generated by a map on an ordered metric space that can be decomposed into increasing and decreasing parts. Our main results provide sufficient conditions for the existence of a globally asymptotically stable fixed point for the map. Applications to discrete-time, stage-structured population models are given. PMID- 16718500 TI - Effects of hyposmotic stress on exocytosis in isolated turbot, Scophthalmus maximus, hepatocytes. AB - The effect of hyposmotic shock on exocytosis was examined in isolated hepatocytes of turbot, a marine flatfish, using the molecular probe FM1-43. Sudden exposure to a reduced osmolality caused an increase in cell exocytic activity related to the osmotic gradient between intra- and extracellular fluids. Cytoskeletal microtubules could contribute to this hyposmotic-induced exocytosis since colchicine inhibited the process. Protein kinase C, phosphatidylinositol-3 kinase, phospholipases A2, C and D could constitute key enzymes in the mechanism since their inhibition by specific agents altered the hyposmotic-induced exocytic activity. Moreover, arachidonic acid and derivates from the 5-lipoxygenase pathway as well as calcium could participate in the process. As regulatory volume decrease (RVD) exhibited by turbot hepatocytes following hyposmotic stimulation involves similar features, a potential role of exocytosis in volume regulation is suggested. In particular, exocytosis could serve RVD by contributing to ATP release since this latter process similarly appeared to be phospholipase D dependent and related to the osmotic gradient. This study provides the first evidence of a volume-sensitive exocytosis that could aim at volume constancy in a marine teleost fish cell type. PMID- 16718501 TI - Vagus nerve stimulation in children less than 5 years old. AB - INTRODUCTION: Vagus nerve stimulation (VNS) has been used in both adults and older children with varying success. MATERIALS AND METHODS: We retrospectively reviewed our experience with VNS in very young children (below 5 years old). The mean age at stimulator implantation was 20.5 months. Two patients were below 2 years old at implantation and two patients were below 1 year old at their initial surgery. The average follow up time for this group was 22 months. RESULTS: Of the six patients (three males and three females) with long-term follow up, 83% had a significant decrease in the frequency of their seizure. Of these, two are seizure free (33%), three are improved (50%), and one (17%) has had no change in seizure status at their most recent clinical examination. Age at implantation of the vagus nerve stimulator did not seem to correlate with patient success. In this group, atonic seizures were found to best respond to VNS with cessation of this type of seizure in two patients. No patients were made worse by the procedure and no morbidity was observed related to VNS. CONCLUSIONS: Based on our small patient cohort, it appears that VNS in very young children with life-threatening epilepsy can be efficacious. Larger groups and other institutional experiences are now needed to verify our findings. PMID- 16718502 TI - Primary combined squamous and small cell carcinoma of the larynx: Report of two cases and discussion of treatment modalities. AB - Combined small cell carcinoma (SMCC) of the larynx consists of SMCC admixed with a component of squamous cell carcinoma or adenocarcinoma. These tumors are very rare and, to date, only a few cases have been fully described. This points out the lack of information available about the correct management of these patients. Here, we describe two additional cases of combined SMCC of the larynx that illustrate the difficulties that we can encounter to diagnose correctly these patients and, by consequence, to treat them adequately. PMID- 16718505 TI - Central adaptations during repetitive contractions assessed by the readiness potential. AB - Physiological fatigue, a loss of maximal force producing capacity, may originate both from changes at the peripheral and at the central level. The readiness potential (RP) provides a measure to study adaptations to physiological fatigue at the motor cortex. We have studied the RP in the course of repetitive contractions at a high force level. Fourteen female healthy subjects made repetitive force grip contractions at 70% of their maximal voluntary contraction (MVC) for 30 min. Contractions were self-paced and inter-squeeze interval was about 7 s. During the repetitive contractions, the area under the curve of the RP almost doubled at electrode Cz and increased fourfold at electrodes C3' and C4'. The onset of negativity moved forward from 1.5 to 1.9 s before force onset at Cz and from 1.0 to 1.6 s and 1.7 s before force onset at C3' and C4', respectively. EMG amplitude and median frequency did not change significantly and MVC after the fatiguing exercise was 93% of MVC before, indicating relatively little physiological fatigue. The increase of the RP during the repetitive contractions is clearly in excess of the almost absent signs of peripheral fatigue. Because the increase of the RP does not lead to an increased force production, we propose that it is a central adaptation counteracting the decrease of cortical efficiency during repetitive contractions. PMID- 16718506 TI - Mechanisms behind the postprandial increase in cardiac output: a clue obtained from transplanted hearts. AB - Consumption of a meal is followed by an increase in cardiac output (CO) which appears to be closely related to the concomitant increase in blood flow to the gastrointestinal organs. To gain information on the mechanism behind this increase in CO we have previously used Doppler ultrasound technique to record circulatory responses to a standardized meal in five patients with recently transplanted and thus denervated hearts. We obtained the surprising result that they reacted to the consumption of a meal with a greater increase in CO than did five matched normal controls. The patients also presented above-normal levels of heart rate (HR) and CO at rest. The same five patients have now been tested 18 months later to re-examine their remarkable cardiac response to ingestion of a meal. The hearts of two patients showed some signs of reinnervation, whereas the hearts of the other three were apparently still fully denervated. However, all five patients once again evolved a marked cardiac response to ingestion of a meal. Postprandial CO reached significantly higher levels in the patients than in the controls. The persistence of such a pronounced postprandial augmentation of CO in transplanted and largely denervated hearts strengthens the assumption that the heart is induced to increase its postprandial performance through the action of a humoral agent of some sort, possibly one of the hormones from the duodenal pancreatic region. PMID- 16718507 TI - Distance of walking in childhood and femoral bone density in perimenopausal women. AB - Kuopio osteoporosis risk factor and prevention (OSTPRE) study is a population based study from Eastern Finland. At baseline in 1989-91, bone densitometry of lumbar spine and femoral neck as assessed by DXA was carried out on women aged 48 58 (n = 3,222). In 1993, menarcheal age and health habits during adolescence were inquired from a postal inquiry. In 1996, a random sample of 254 women who had been premenopausal at baseline was interviewed over phone. They were asked how many kilometers per day they had walked to school and back, in each grade of primary school. The study sample (N = 185) was formed by excluding women with menarcheal age of >14 or <11 years. Women with any reported HRT history were also excluded. The mean age of the study sample was 50.7 (1.63) years, weight 70.8 (13.1) kg, height 161.5 (5.0) cm, and mean walking distance to and fro from the school at ages from 9 to 11 years was 2.7 (1.7) km. In regression analysis, the walking distance was associated with femoral BMD (r = 0.18, P = 0.015). After adjusting for baseline age, weight and height, this association persisted (P = 0.025). When walking distance was categorized as I = 0-0.549 km, II = 0.55-1.99 km, III = 2.0-4.99 and IV = 5.0 km and more, the respective means for femoral BMDs were 0.92, 0.97, 0.98 and 1.01 g/cm(2). Statistical significance persisted after adjusting for height, weight, age, grip strength, calcium intake, smoking, place of residence, use of contraceptive pills, physical load of work and baseline physical activity (P = 0.032). A 10 year follow-up revealed no changes in bone loss rate between the groups and femoral BMD benefits persisted (repeated measures analysis = NS). Walking distance was not associated with spinal bone density. Even though walking is a low impact activity, walking before menarche may have a moderate but long-lasting positive effect on femoral peak bone density. Lack of walking and similar low impact physical activities during peak growth years may have a negative effect on peak bone mass formation. PMID- 16718508 TI - Age-related fatigability of the ankle dorsiflexor muscles during concentric and eccentric contractions. AB - This study compares the fatigability of the ankle dorsiflexors during five sets of 30 maximal concentric and eccentric contractions in young and elderly adults. The torque produced by the ankle dorsiflexors and the average surface electromyogram (aEMG) of the tibialis anterior were continuously recorded. The contribution of central and peripheral mechanisms to muscle fatigue was tested before, after each set of contractions, and during a 30 min recovery period by the superimposed electrical stimulation method. The compound muscle action potential (M-wave), the mechanical response to single (twitch) and paired (doublet) stimulation, and the postactivation potentiation were also recorded. Compared with young subjects, elderly adults exhibited a greater loss of torque for concentric (50.2 vs. 40.9%; P<0.05) and eccentric (42.1 vs. 27.1%; P < 0.01) contractions. Although young subjects showed a lesser decrease in torque during the eccentric compared with concentric contractions, elderly adults experienced similar fatigability for the two types of contractions despite a comparable depression in the EMG activity of both groups and contraction types (10-20%). As tested by the interpolated-twitch method and aEMG/M-wave ratio, voluntary activation was not altered during either type of contraction or for either age group. During the two fatigue tasks, only elderly adults experienced a decrease in M-wave area (26.4-35.4%; P < 0.05). All together, our results suggest that the fatigue exhibited by both young and elderly adults during maximal concentric and eccentric contractions mainly involved peripheral alterations and that elderly adults may also have experienced a decline in neuromuscular propagation. PMID- 16718510 TI - Is the psychological refractory period effect for ideomotor compatible tasks eliminated by speed-stress instructions? AB - It has been argued that the psychological refractory period (PRP) effect is eliminated with two ideomotor compatible tasks when instructions stress fast and simultaneous responding. Three experiments were conducted to test this hypothesis. In all experiments, Task 1 required spatially compatible manual responses (left or right) to the direction of an arrow, and Task 2 required saying the name of the auditory letter A or B. In Experiments 1 and 3, the manual responses were keypresses made with the left and right hands, whereas in Experiment 2 they were left-right toggle-switch movements made with the dominant hand. Instructions that stressed response speed reduced reaction time and increased error rate compared to standard instructions to respond fast and accurately, but did not eliminate the PRP effect on Task 2 reaction time. These results imply that, even when response speed is emphasized, ideomotor compatible tasks do not bypass response selection. PMID- 16718509 TI - Expression and activity of the glutamate transporter EAAT2 in cardiac hypertrophy: implications for ischaemia reperfusion injury. AB - The expression and activity of the glutamate transporter, excitatory amino acid transporter 2 (EAAT2), in cardiac hypertrophy were investigated with respect to glutamate's potential as a cardioprotective agent. Sarcolemmal vesicles (SV) isolated from hypertrophic hearts of male spontaneously hypertensive rats (SHR) or normotrophic hearts from age-matched male Wistar Kyoto rats (WKY) were used to measure the relative level of EAAT2 expression by Western blotting and the initial rate of 0-0.3 mM L-[(14)C]glutamate uptake. The effects of 20-min global normothermic ischaemia +/-0.5 mM glutamate on cardiac function were measured in isolated working SHR/WKY hearts. In a separate series of hearts, glutamate, lactate and ATP levels were measured. Both the level of EAAT2 expression and the V (max) for sodium-dependent L-[(14)C]glutamate uptake were significantly greater in SHR SV compared to WKY SV. The reperfusion cardiac output (CO) of SHR hearts was significantly worse than that of the WKY hearts (24.3+/-2.2 ml/min vs 39.8+/ 3.3 ml/min, n=7/9+/-SE, p<0.01). The addition of 0.5 mM L-glutamate improved the SHR reperfusion CO to 45.2+/-5 ml/min, (n=6+/-SE, p<0.01) but had no effect on WKYs (46.2+/-3.8 ml/min, n=6+/-SE). SHR with 0.5 mM L-glutamate had higher glutamate levels at the start of ischaemia, plus higher glutamate and ATP levels at the end of ischaemia compared to any other group. These results suggest that increased glutamate transporter expression and activity in the SHR hearts helped facilitate glutamate entry into the SHR cardiomyocytes leading to improved myocardial metabolism during ischaemia and better functional recovery on reperfusion. PMID- 16718511 TI - Effect of colour pop-out on the recognition of letters in crowding conditions. AB - The crowding effect of adjacent objects on the recognition of a target can be reduced when target and flankers differ in some feature, that is irrelevant to the recognition task. In this study, the mechanisms of this effect were explored using targets and flankers of the same and different colours. It was found that facilitation nearly equal to that of differently coloured targets and flankers can be observed with a differently coloured background blob in the location of the target. The different-colour effect does not require advance knowledge of the target and flanker colours, but the effect increases in the course of three trials with constant mapping of colours. The results are consistent with the notion of exogenous attention that facilitates the processing at the most salient locations in the visual field. PMID- 16718512 TI - Neospora caninum immunoblotting improves serodiagnosis of bovine neosporosis. AB - Neospora caninum ranges among the major causes of infectious abortion in cattle worldwide. The present study was designed to improve the serodiagnostic tools by complementing a conventional ELISA with a highly sensitive and species-specific N. caninum immunoblot. To evaluate this test combination, sera from several groups of cows were tested. The first group, consisting of experimentally infected calves, showed that immunoblot antibody reactivities were detectable 1 to 3 days earlier than those found in ELISA. The first immunodominant bands that appeared were a 29-kDa (NcSAG1) and a 36-kDa (NcSRS2) antigen. Other groups, based upon naturally infected cattle, were used to compare the diagnostic sensitivity of ELISA and immunoblotting. Overall, N. caninum immunoblotting exhibited a higher sensitivity (98%) than ELISA (87%). Conversely, immunoblotting also confirm in two other cases, true transient negativation in some animals. In general, banding patterns and band staining intensity correlated to the semiquantitative ELISA findings. On the other hand, the banding pattern could not be used to discriminate between sera from animals with a recent abortion and those of cows with latent N. caninum infection. We also addressed putative cross reactions due to infection with Toxoplasma gondii. Sera from animals with a serologically proven T. gondii infection were either clearly negative by Neospora immunoblotting or they yielded a specific immunoblot antibody profile indicating a double infection with N. caninum. Sera from animals with positive findings in both Toxoplasma and Neospora ELISA thus provided dichotomic results in the immunoblot by allowing to confirm or to rule out the specificity of the antibody reaction in Neospora ELISA. Altogether, our findings demonstrate that N. caninum immunoblotting is a very sensitive and specific complementary tool to improve the serology for N. caninum infections in cattle. PMID- 16718513 TI - [Clinical pathways--practical aid for the physician? Example: clarification of the suspicion of tuberculosis]. AB - Guidelines, clinical pathways and clinical algorithms are popular instruments nowadays to ensure quality as well as the economic efficiency of medical work. These terms themselves, however, are frequently defined only in a diffuse way. Thus, medical standard procedures often complicate clinical workflows more than to facilitate decision making in everyday life. In our department, feasible standardized approaches have been generated in the form of structured text documents, which on the one hand can aid clinical decision making at the bedside and on the other hand serve as medical sketches for the generation of operational treatment paths on an interdisciplinary level. Structure and content of such an instructional text are exemplified here using our standardized document for the diagnostic approach when tuberculosis is suspected. PMID- 16718517 TI - A psychiatric mother-baby day hospital for pregnant and postpartum women. AB - Major depression and other psychiatric disorders are common during pregnancy and the postpartum period, yet these disorders remain largely under-diagnosed and under-treated. Developing programs that are uniquely tailored to meet the needs of perinatal psychiatric patients can improve both the quality and acceptability of care. In this report, we describe the development and implementation of a novel mother-baby day hospital service designed to meet the mental health needs of this special population, and present preliminary data regarding treatment acceptability and effectiveness. Our experience using this model of care for the past five years has suggested that specialized units such as this one represent an acceptable, effective, fiscally viable approach to the care of pregnant and postpartum psychiatric patients. Further research is needed to more thoroughly assess the effectiveness of this type of specialized perinatal service. PMID- 16718515 TI - Requirements regarding dose rate and exposure time for killing of tumour cells in beta particle radionuclide therapy. AB - PURPOSE: The purpose of this study was to identify combinations of dose rate and exposure time that have the potential to provide curative treatment with targeted radionuclide therapy applying low dose rate beta irradiation. METHODS: Five tumour cell lines, U-373MG and U-118MG gliomas, HT-29 colon carcinoma, A-431 cervical squamous carcinoma and SKBR-3 breast cancer, were used. An experimental model with 10(5) tumour cells in each sample was irradiated with low dose rate beta particles. The criterion for successful treatment was absence of recovery of cells during a follow-up period of 3 months. The initial dose rates were in the range 0.1-0.8 Gy/h, and the cells were continuously exposed for 1, 3 or 7 days. These combinations covered dose rates and doses achievable in targeted radionuclide therapy. RESULTS: Continuous irradiation with dose rates of 0.2-0.3 and 0.4-0.6 Gy/h for 7 and 3 days, respectively, could kill all cells in each tumour cell sample. These treatments gave total radiation doses of 30-40 Gy. However, when exposed for just 24 h with about 0.8 Gy/h, only the SKBR-3 cells were successfully treated; all the other cell types recovered. There were large cell type-dependent variations in the growth delay patterns for the cultures that recovered. The U-118MG cells were most resistant and the U-373MG and SKBR-3 cells most sensitive to the treatments. The HT-29 and A-431 cells were intermediate. CONCLUSION: The results serve as a guideline for the combinations of dose rate and exposure time necessary to kill tumour cells when applying low dose rate beta irradiation. The shift from recovery to "cure" fell within a narrow range of dose rate and exposure time combinations. PMID- 16718520 TI - Concurrent craniospinal radiotherapy and intrathecal chemotherapy in patient with acute promyelocytic leukemia second relapsed in central nervous system (CNS) following allogeneic stem cell transplantation. PMID- 16718519 TI - Extension arm facilitated PEGylation of hemoglobin: correlation of the properties with the extent of PEGylation. AB - Human hemoglobin (Hb) conjugated with six copies of PEG-5K is nonhypertensive. The hexaPEGylated Hb exhibits molecular size homogeneity in spite of the chemical heterogeneity with respect to the sites of conjugation (Manjula et al., 2005). In the present study, Hb conjugated with an average of 4, 6, 8 and 10 copies of PEG 5K chains have been generated using the extension arm facilitated PEGylation protocol. Except for the tetraPEGylated Hb, all the other products exhibit molecular size homogeneity. The molecular, colligative and functional properties of PEG-Hb conjugates have been correlated with the extent of PEGylation. The results imply that six copies of PEG-5K chains are accommodated on Hb without significant crowding on the molecular surface. As more copies of PEG-5K chains are conjugated to form octa and deca PEGylated Hb, the PEG-chains conjugated appear to undergo transition from a mushroom (compact) to a brush-like conformation (extended conformation) with a concomitant decrease in the propensity of the molecule to transition from oxy to deoxy conformation in the presence of allosteric effectors. The viscosity and the colloidal osmotic pressure of Hb increase with the number of the PEG-chains conjugated in an exponential fashion. The composition of the PEGylated Hb generated appears to be controlled by (i) high reactivity of thiol groups of the extension arms on Hb with maleimide-PEG, (ii) increase in the viscosity of the reaction mixture as the level of PEGylation increases and (iii) increased resistance induced by the PEG shell of PEGylated Hb to accommodate more PEG-chains as the level of PEGylation increases. Potential implications of extent of PEGylation on the oxygen delivery by PEG-Hb conjugate in vivo have been discussed. PMID- 16718521 TI - Mitotic activity of multinucleated giant cells with glial fibrillary acidic protein immunoreactivity in glioblastomas: an immunohistochemical double labeling study. AB - To investigate the mitotic activity of multinucleated giant cells (MNGCs) with glial fibrillary acidic protein (GFAP) in glioblastomas, double immunohistochemical staining for GFAP and Ki67 was performed in formalin-fixed and paraffin-embedded specimens obtained from 12 primary glioblastomas with MNGCs including three giant cell glioblastomas. The Ki67 labeling index (LI:%) of GFAP+ tumor cells ranged from 0 to 5.6 (2.5+/-1.7, mean+/-standard deviation). The Ki67 LI of GFAP- tumor cells ranged from 18.6 to 35.9 (24.7+/-6.6). The Ki67 LI of GFAP+ cells was significantly lower than that of GFAP- cells (P<0.0001). The number of Ki67+ GFAP- MNGCs ranged from 0 to 23 (8.6+/-8.2). The number of Ki67- GFAP+ MNGCs ranged from 0 to 15 (6.2+/-5.1). There was no significant difference between them (P=0.42). The Ki67 LI of GFAP+ MNGCs was 10.4+/-12.6. The Ki67 LI of GFAP- MNGCs was 45.9+/-29.9. The Ki67 LI of GFAP+ MNGCs was significantly lower than that of GFAP- MNGCs (P<0.01). Our study suggested that MNGCs with mitotic capacity were not dominant and that GFAP+ MNGCs had little mitotic activity in glioblastomas. MNGCs identified in glioblastomas may develop via not only the proliferation of abnormal nuclei in a single tumor cell but also other processes. PMID- 16718522 TI - Technical hurdles in a pilot clinical trial of combined B7-2 and GM-CSF immunogene therapy for glioblastomas and melanomas. AB - OBJECTIVE: Malignant glioblastomas and melanomas continue to have a dismal prognosis despite advances in conventional therapy. This has led to investigations of novel treatment strategies including immunogene therapy. We report a pilot clinical trial of combined B7-2 and GM-CSF immunogene therapy for gliomas and melanomas and discuss technical hurdles encountered. METHODS: Patients with recurrent malignant gliomas or medically refractory melanomas were vaccinated with irradiated autologous tumor cells transduced with B7-2 and GM-CSF genes using a retroviral vector. Patients were monitored for toxicity, inflammatory/immune reactions, and clinical status. RESULTS: Vaccine preparation was attempted from 116 malignant glioma and 32 melanoma specimens. Adequate vaccines could only be prepared for five glioblastoma and three melanoma patients. Six patients (three recurrent glioblastomas and three melanomas) were actually vaccinated. Minor toxicities included flu-like symptoms (3/6), injection site erythema (4/6), and asymptomatic elevations in liver enzymes (3/6). Most patients showed evidence of an inflammatory response but specific anti-tumor immunity was not demonstrated. All six patients have died, although three patients with minimal residual disease at treatment had prolonged recurrence-free intervals after vaccination. CONCLUSIONS: Combined B7-2 and GM-CSF immunogene therapy for glioblastomas and melanomas using autologous tumor cells has many technical pitfalls hindering large scale application and evaluation. As a result, this pilot study was too limited to draw meaningful conclusions regarding safety or anti-tumor immunity. While immunotherapy has been promising in pre-clinical studies, alternate strategies will be required to bring these benefits to patients. PMID- 16718535 TI - Presenting symptoms of nonerosive and erosive esophagitis in pediatric patients. AB - Children and adolescents with symptomatic gastroesophageal reflux disease (GERD) and erosive esophagitis (EE) of grade >/=2 (n=45) or nonerosive esophagitis (NEE) (n=45) were assessed to determine the relationship between presenting symptoms, esophagitis severity, and patient age. Overall, regurgitation/vomiting, abdominal pain, and cough were the most frequent symptoms. The prevalence and severity of anorexia/feed refusal was significantly greater in EE versus NEE children; this symptom was also significantly more prevalent in younger (1-5 years) children (both NEE and EE groups) compared to older children. Cough was significantly less severe in NEE adolescents than in younger children. Cough, anorexia/feed refusal, and regurgitation/vomiting were more severe and heartburn was less severe in EE children aged 1-5 years compared with older patients. In conclusion, GERD in children manifests differently than that in adults and symptoms vary with patient age. Symptoms were not predictive of presence or lack of mucosal damage. PMID- 16718536 TI - Effect of physician training on fluoroscopy time during ERCP. AB - Our purpose was to compare fluoroscopy time during endoscopic retrograde cholangiopancreatography (ERCP) between endoscopists with different levels of experience. We performed a cross-sectional analysis of 269 consecutive ERCPs at an academic hospital during 1 year. Median fluoroscopy time was significantly longer in more complex cases, such as in therapeutic (406.5 s [IQR, 235.5-685]) compared to diagnostic (202 s [IQR, 141-481]; P = 0.002) procedures. The experience (number of prior ERCPs) of gastroenterology fellows involved in procedures was an independent predictor of shorter fluoroscopy time when controlling for patient and procedure characteristics (P < 0.0001). Median fluoroscopy time was 2.73 min shorter after at least 50 procedures had been performed (P = 0.039). Time for ERCPs involving fellows was not significantly longer than cases by attending physicians alone (P = 0.23). Increased experience is associated with lower radiation exposure during ERCP training. Radiation reduction methods should be prospectively investigated and integrated into training programs. PMID- 16718538 TI - Reflections on the evaluation of a Cambodian youth dance program. AB - Evaluating a youth program whose goals are to provide instruction in Cambodian dance, increase awareness and pride in Cambodian culture, promote healthy behaviors, and create linkages within the community has been a challenge. A primary source of conflict was incorporating evaluation methods that were required of all funded programs with our own specifically tailored measures. One of our concerns was that the required tools were not culturally appropriate for our participants. Our experiences reinforce the importance of forming partnerships that embrace principles of respect, equity, and empowerment among all involved before establishing a research agenda. The choices we made and did not make contributed to our struggles and frustration and also to the insight that was gained. Our analysis examines the importance of clear communication, cultural awareness, tailoring evaluation, and meaningful participation. We believe that the lessons we learned will help facilitate the conduct of culturally sensitive community-based research. PMID- 16718539 TI - Links between antisocial behavior and depressed mood: the role of life events and attributional style. AB - Comorbidity between antisocial behavior and depression in adolescence is widely recognized. This paper examines whether links with depressed mood differ among three subtypes of antisocial behavior: oppositionality, physical aggression and delinquency. In addition we examine two possible contributors to these links: negative life events that are dependent upon the individual's actions and depressogenic attributional style. Data are drawn from the G1219 large-scale community twin and sibling sample, and include 2409 questionnaire responses from youths aged between 13 and 19 living in the United Kingdom. Depressed mood was independently associated with both oppositionality and delinquency, but not with physical aggression. Dependent negative life events were strongly implicated in the association between delinquency and depressed mood, whereas depressogenic attributional style was implicated in the associations of both oppositionality and delinquency with depressed mood. Oppositionality remained a significant predictor of depressed mood after accounting for attributional style and life events whereas delinquency did not. The pattern of associations was largely similar in boys and girls. We discuss these results in terms of developmental models of the links between antisocial behavior and depressed mood. PMID- 16718540 TI - Does the quality of prenatal care matter in promoting skilled institutional delivery? A study in rural Mexico. AB - OBJECTIVES: To determine if the quality of prenatal care predicts skilled institutional delivery, a primary means of reducing maternal mortality. METHODS: The probability of skilled institutional delivery is predicted among 4173 rural low-income women of reproductive age in seven Mexican states, as a function of maternal retrospective reports about prenatal care services received in 1997 2003. RESULTS: Women who received most prenatal care procedures were more likely to have a skilled institutional delivery (OR 2.29, 95% CI 1.18, 4.44). Women who received less than the 75th percentile of prenatal care procedures were not significantly different from those who received no prenatal care. CONCLUSIONS: Policies promoting increased access to prenatal services should be linked to the promotion of practice standards to impact health and behavioral outcomes. PMID- 16718542 TI - Effects of family risk factors on dosage and efficacy of a family-centered preventive intervention for rural African Americans. AB - This study was designed to test hypotheses about family risk factors and their links to dosage and efficacy of a family-centered preventive intervention. Participants were 172 families with an 11 year-old child randomly assigned to the intervention condition in the Strong African American Families Program (SAAF). Two family risk factors, ratio of adults to children in the household and youth unconventionality, were negatively related to dosage, defined as number of intervention sessions attended. Dosage, in turn, was associated with changes in targeted parenting behavior across the 7 months between pretest and posttest. The effect of family risk factors on the link between program dosage and changes in parenting behavior was stronger for families experiencing more risks. The results highlight the need for engagement strategies for recruiting and retaining high risk families in preventive interventions. PMID- 16718555 TI - Chemically mediated species recognition in closely related Podarcis wall lizards. AB - In many animals, chemical signals play an important role in species recognition and may contribute to reproductive isolation and speciation. The Iberian lizards of the genus Podarcis, with up to nine currently recognized lineages that are often sympatric, are highly chemosensory and provide an excellent model for the study of chemically mediated species recognition in closely related taxa. In this study, we tested the ability of male and female lizards of two sister species with widely overlapping distribution ranges (Podarcis bocagei and P. hispanica type 1) to discriminate between conspecific and heterospecific mates by using only substrate-borne chemical cues. We scored the number of tongue flicks directed at the paper substrate by each individual in a terrarium previously occupied by a conspecific or a heterospecific lizard of the opposite sex. Results show that males of P. bocagei and P. hispanica type 1 are capable of discriminating chemically between conspecifics and heterospecifics of the opposite sex, but females are not. These results suggest that differences in female, but not male, chemical cues may underlie species recognition and contribute to reproductive isolation in these species. The apparent inability of females to discriminate conspecific from heterospecific males, which is not because of reduced baseline exploration rates, is discussed in the context of sexual selection theory and species discrimination. PMID- 16718556 TI - Kairomonal response of predators to three pine bast scale sex pheromones. AB - The kairomonal activity of the sex pheromones of three pine bast scales, Matsucoccus feytaudi, Matsucoccus josephi, and Matsucoccus matsumurae, as well as a new analog of the M. feytaudi sex pheromone, were investigated in pine forests of France, Portugal, and Italy. The response of the maritime pine bast scale predators, Elatophilus spp. and Hemerobius stigma, was used to test the influence of trapping methods, kairomone composition, and dose. Both predators showed significant attraction to all compounds except to the sex pheromone of M. josephi. Significant increase in captures was observed as a function of dose, and within the studied dose range, up to 2200 mug, no threshold saturation limits were observed for any of the attractive compounds. Trap design and size did not significantly influence predator captures, except for high population levels of Elatophilus crassicornis, when plate traps were more efficient than delta traps. Geographic variations were found in the kairomonal responses patterns of both predators, with the M. matsumurae sex pheromone being more attractive to the oriental populations from Corsica and Italy, whereas the western populations in Aquitaine and Portugal were more attracted to the M. feytaudi sex pheromone. PMID- 16718557 TI - Field evidence of an airborne enemy-avoidance kairomone in wolf spiders. AB - Hogna helluo, Pardosa milvina, and Trochosa ruricola are co-occurring species of wolf spiders within agricultural fields in the eastern USA. The largest species, H. helluo, is a common predator of the two smaller species, P. milvina and T. ruricola. H. helluo frequently resides within soil fissures where P. milvina and T. ruricola may be attacked when they enter or walk near these fissures. We tested the ability of P. milvina and T. ruricola to avoid H. helluo-containing burrows by detecting airborne enemy-avoidance kairomones associated with H. helluo. To simulate soil fissures and control for visual and vibratory means of predator detection, we baited funneled pitfall traps with one of the following (N = 20 traps/treatment): (1) blank (empty trap); (2) one house cricket (Acheta domesticus); (3) one adult female H. helluo; and (4) one adult male H. helluo. Over two separate 3-d periods, we measured pitfall capture rates of P. milvina and T. ruricola as well as other incidentally captured ground-dwelling arthropods. During the day, male P. milvina showed significant avoidance of pitfall traps baited with H. helluo of either sex but showed no avoidance of empty traps or those containing crickets. At night, male T. ruricola showed a qualitatively similar pattern of avoiding H. helluo-baited traps, but the differences were not statistically significant. We found no evidence that other ground-dwelling arthropods either avoided or were attracted to H. helluo-baited traps. This study suggests that an airborne enemy-avoidance kairomone may mediate behavior among male P. milvina in the field. PMID- 16718558 TI - Hexyl decanoate, the first trail pheromone compound identified in a stingless bee, Trigona recursa. AB - Foragers of many species of stingless bees guide their nestmates to food sources by means of scent trails deposited on solid substrates between the food and the nest. The corresponding trail pheromones are generally believed to be produced in the mandibular glands, although definitive experimental proof has never been provided. We tested the trail following behavior of recruits of Trigona recursa in field experiments with artificial scent trails branching off from natural scent trails of this stingless bee. First-time recruits (newcomers) did not follow these trails when they were laid with pure solvent or mandibular gland extract. However, they did follow trails made with labial gland extract. Chemical analyses of labial gland secretions revealed that hexyl decanoate was the dominant component (72.4 +/- 1.9% of all volatiles). Newcomers were significantly attracted to artificial trails made with synthetic hexyl decanoate, demonstrating its key function in eliciting scent-following behavior. According to our experiments with T. recursa, the trail pheromone is produced in the labial glands and not in the mandibular glands. Hexyl decanoate is the first component of a trail pheromone identified and proved to be behaviorally active in stingless bees. PMID- 16718559 TI - Evidence for four-component close-range sex pheromone in the parasitic wasp Glyptapanteles flavicoxis. AB - Females of the parasitic wasp Glyptapanteles flavicoxis (Hymenoptera: Braconidae) deposit a close-range sex pheromone from their abdominal tip that attracts conspecific males and elicits wing-fanning behavior. In this study, we isolated the pheromone components and determined their role in the males' behavior. In coupled gas chromatographic-electroantennographic detection (GC-EAD) analyses of the females' body extract, four components (below GC detection) elicited strong responses from male antennae. Monitored by GC-EAD, the components were separated by flash silica gel and high-performance liquid chromatography. Y-tube olfactometer experiments with one or more components revealed that all are necessary to elicit short-range attraction and wing-fanning responses by males. These components remained below detection threshold of the mass spectrometer (approximately 10 pg) even when 4500 female equivalents were analyzed in a single injection, which attests to the potency of the pheromone and the insects' sensitivity to it. PMID- 16718560 TI - Orientational behaviors and EAG responses of male codling moth after exposure to synthetic sex pheromone from various dispensers. AB - The effect of brief pheromone exposures on responses of codling moth (Cydia pomonella L.) males was tested by flight-tunnel and electroantennogram (EAG) studies. Males were preexposed to pheromone for up to 3 min as they sat in release cages or for shorter times (a few seconds to several min) upon initiating flights or orienting in plumes. Brief exposures to Isomate-C Plus dispensers nearly eliminated moth orientations to 0.1 mg codlemone [(E,E)-8,10-dodecadien-1 ol] and 0.1 mg three-component (codlemone/12OH/14OH, 100:20:5 ratio) lures 15 min later. However, there was no associated change in EAG responses between preexposed and control moths. Behavioral responses of Isomate-C Plus-exposed males were normal 24 hr following exposure. The reduced sexual responsiveness observed following exposure to Isomate dispensers appeared to be associated with an elevation of response threshold. Brief preexposure to 0.1 mg codlemone and three-component lures also reduced orientational behavior of males 15 min later, but to a lesser degree than when preexposed to Isomate-C Plus dispensers. Male behavior following preexposure to a 0.1 mg codlemone/pear ester [(2E,4Z)-2,4 decadienoate] lure (1:1 ratio) was no different from exposure to codlemone only. Orientational disruption in plots treated with 10 dispensers of Isomate-C Plus per tree was 88.3 and 95.9% for 1.0 and 0.1 mg codlemone lures, respectively. Some males did orient to 0.1 mg codlemone lures so we caution that flight-tunnel experiments on preexposure may overestimate the actual pheromone exposure dosage received by feral moths in treated orchards. Importantly, this work documents that a portion of feral males within a population has the capacity to overcome communicational disruption by high densities of Isomate-C Plus dispensers. PMID- 16718561 TI - Predator-labeling effect on byssus production in marine mussels Perna viridis (L.) and Brachidontes variabilis (Krauss). AB - Mussels Perna viridis and Brachidontes variabilis were exposed to chemical cues from the predatory crab Thalamita danae maintained on different diets, and byssal thread production of the mussels was studied. P. viridis produced the highest number as well as the thickest and longest byssal threads when they were exposed to crabs maintained on a diet of P. viridis as compared with those exposed to crabs maintained on a diet of the top shell Monodonta labio, the rock oyster Saccostrea cucullata, or crabs that were starved. For B. variabilis, results were similar, in that a diet containing B. variabilis elicited the greatest response as compared with other treatments. This indicates that the mussels were able to discriminate chemical cues released from predators maintained on different diets, and respond accordingly to the level of predation risk. By increasing the strength of byssal attachment as a defensive trait, the chance of being dislodged and consumed by crabs is reduced. As energy cost involved in the induction of an antipredatory response is considerable, this defensive trait seems to be an advantage to the mussels in enhancing efficiency. The short response time in byssal thread production allows the mussels to increase resistance against predation by crabs at the time when predation pressure is the highest in a tidal cycle. PMID- 16718562 TI - Chemical defenses of cryptic and aposematic Gastropterid molluscs feeding on their host sponge Dysidea granulosa. AB - Numerous opisthobranchs are known to sequester chemical defenses from their prey and use them for their own defense. Information on feeding biology is critical for understanding the ecology and evolution of molluscs, yet information on feeding biology is still scarce for many groups. Gastropterid molluscs are often found on sponges, but there is controversy as to whether they are true sponge feeders. On Guam, we found the gastropterids Sagaminopteron nigropunctatum and S. psychedelicum on the sponge Dysidea granulosa. They seem to rely on contrasting defense strategies as S. psychedelicum has vivid colors, consistent with the warning coloration found in many chemically defended opisthobranchs, whereas S. nigropunctatum is highly cryptic on the sponge. S. nigropunctatum is avoided by the pufferfish Canthigaster solandri in aquarium assays. We analyzed the secondary metabolites of the two species and found that both share polybrominated diphenyl ethers (BDEs) with their host sponge D. granulosa. S. psychedelicum and S. nigropunctatum sequester the major BDE in the sponge and accumulate it in the mantle at approximately the same concentration as in the sponge (4.03 and 2.37%, respectively), and concentrate it in their parapodia at over twice the sponge concentration (7.97 and 10.10%, respectively). We also detected trace amounts in the mucus secretion of S. psychedelicum, and quantified significant amounts in the mucus (1.84%) and egg masses (2.22%) of S. nigropunctatum. Despite contrasting color patterns displayed by the two gastropterid species, they seem to share a similar chemical defense strategy, i.e., they feed on D. granulosa and accumulate the major BDE of the sponge in their tissues. PMID- 16718563 TI - Selected ectomycorrhizal fungi of black spruce (Picea mariana) can detoxify phenolic compounds of Kalmia angustifolia. AB - Allelopathy has been implicated as a factor contributing toward failure of black spruce (Picea mariana) regeneration in Kalmia angustifolia-dominated sites in eastern Canada. Several phenolic acids of Kalmia origin inhibit primary root growth of black spruce. We tested the hypothesis that some well-adapted conifer ectomycorrhizae can degrade and detoxify water-soluble phenolic compounds produced by Kalmia and use the degraded products as a carbon source to stimulate growth. We found that hyphal growth of Paxillus involutus, a common ectomycorrhizal fungus of black spruce, was stimulated by water leachates of Kalmia leaf and litter. An equimolar mixture of three phenolic acids (ferulic, o coumaric, and o-hydroxyphenylacetic acid), commonly found in Kalmia, had no negative effects on fungal growth at 1 mM concentration. The o hydroxyphenylacetic (o-HPA) acid, which is known to be toxic to black spruce, was found to stimulate the growth of Laccaria laccata, L. bicolor, and P. involutus (isolates 211804 and 196554) by 38.4, 29.3, 25.0, and 18.9%, respectively, at 1 mM. Pure ferulic, o-coumaric, and o-HPA acids were degraded by 100, 98, and 79.5%, respectively, within 10 d in the presence of P. involutus 211804. However, L. laccata could not tolerate high concentrations of the Kalmia leachates. P. involutus and L. bicolor used o-HPA acid as a carbon source when cultured in noncarbon nutrient medium. The 0.5 and 0.2 mM o-HPA acid inhibited the root growth of black spruce. However, after solutions had been exposed to a culture of P. involutus, they had no significant effect on seedling growth of black spruce. We concluded that some ectomycorrhizal fungi, such as P. involutus and L. bicolor, are able to degrade Kalmia phenolics. Our findings point to a mechanism by which ectomycorrhizal species can control species interactions in higher plants by changing the rhizosphere chemistry. PMID- 16718564 TI - Effects of elevated carbon dioxide and ozone on foliar proanthocyanidins in Betula platyphylla, Betula ermanii, and Fagus crenata seedlings. AB - Proanthocyanidins (PAs) or condensed tannins are a major group of phenolic compounds in the leaves of birch trees and many other woody and herbaceous plants. These compounds constitute a significant allocation of carbon in leaves and are involved in plant responses to environmental stress factors, such as pathogens or herbivores. In some plants, PA concentrations are affected by atmospheric carbon dioxide (CO(2)) and ozone (O(3)) levels that may influence, for example, species fitness, community structure, or ecosystem nutrient cycling. Therefore, a study on the quantitative response of PAs to elevated concentrations of carbon dioxide (CO(2)) and ozone (O(3)) was undertaken in seedlings of Betula platyphylla, Betula ermanii, and Fagus crenata. Seedlings were exposed to ambient or elevated O(3) and CO(2) levels during two growing seasons in the Kanto district in Japan. Ten open-top chambers were used for five different treatments with two replicates: filtered air (FA), ambient air (ambient O(3), 43 ppb; ambient CO(2), 377 ppm), elevated O(3) (1.5 x ambient O(3), 66 ppb), elevated CO(2) (1.5 x ambient CO(2), 544 ppm), and elevated O(3) and CO(2) combined. In addition, seedlings growing in natural conditions outside of chambers were studied. Leaf samples were analyzed for total PA concentrations by butanol-HCl assay and for polymeric PA concentrations by normal-phase high-performance liquid chromatography. Total PA concentrations in leaves of all species were similarly affected by different treatments. They were significantly higher in seedlings treated with elevated CO(2) and O(3) combined, and in seedlings growing outside chambers compared with the FA controls. F. crenata contained only traces of polymeric PAs, but significant species x treatment interaction was observed in the polymeric PA concentrations in B. ermanii and B. platyphylla. In B. platyphylla, leaves treated with elevated CO(2) + O(3) differed significantly from all other treatments. It was suggested that the strongest effect of elevated CO(2) and O(3) combined on leaf PA contents resulted from the additive effect of these environmental factors on phenolic biosynthesis. PMID- 16718565 TI - Oviposition in Delia platura (Diptera, Anthomyiidae): the role of volatile and contact cues of bean. AB - The choice of a suitable oviposition site by female insects is essential for survival of their progeny. Both olfactory and contact cues of the oviposition site may mediate this choice. The polyphagous Delia platura (Diptera: Anthomyiidae), a severe agricultural pest of numerous crops, lays eggs in the soil close to germinating seeds. Maggots feed upon the cotyledons. Only little is known about the cues guiding oviposition behavior. In this study, the effects of both olfactory and contact cues of beans (Phaseolus vulgaris) on oviposition of D. platura females were tested. Egg deposition on germinated beans was preferred to egg deposition on ungerminated beans or on beans in different postgerminating developmental stages. Olfactory cues of germinating beans alone stimulated female flies to lay eggs. Additional contact cues of germinating beans seemed to enhance the response, but the difference was not significant. Surface extracts of germinating beans sprayed on surrogate beans showed that both polar and nonpolar substances stimulated oviposition of D. platura flies. Gas chromatography electroantennographic detection recordings of head space samples of germinating beans showed positive response of females to different compounds. We conclude that olfaction plays a major role when D. platura females are searching for oviposition sites. Volatile compounds released from germinating beans such as 4 hydroxy-4-methyl-2-pentanone, 1-hepten-3-one, 1-octen-3-ol, and 3-octanone should be considered as key compounds that mediate oviposition behavior. The use of different sensory modalities by closely related species of Delia is discussed. PMID- 16718566 TI - Early herbivore alert: insect eggs induce plant defense. AB - Plants are able to "notice" insect egg deposition and to respond by activating direct and indirect defenses. An overview of these defenses and the underlying mechanisms is given from a tritrophic perspective. First, the interface between plant and eggs is addressed with respect to the mode of attachment of eggs on the plant surface. It is elucidated which plant cells might respond to components from insect eggs or the egg deposition. The scarce knowledge on the elicitors associated with the eggs or the egg-laying female is outlined. Since endosymbiotic microorganisms are often present on the eggs, and microorganisms are also abundant on the leaf surface, the role of these hidden players for eliciting oviposition-induced plant responses is considered. Furthermore, the question of which physiological and molecular processes are induced within the plant in response to egg deposition is addressed. Second, studies on the response of the herbivorous insect to oviposition-induced plant defenses are outlined. Third, the importance of oviposition-induced plant volatiles and contact cues for host and prey location of parasitoids and predators is discussed in the context of other informative chemicals used by carnivores when searching for food. Finally, physiological and ecological costs of oviposition-induced plant responses are addressed. PMID- 16718567 TI - Effects of genetic modification on herbivore-induced volatiles from maize. AB - Large-scale implementation of transgenic crop varieties raises concerns about possible nontarget effects on other organisms. This study examines the effects of genetic modification on plant volatile production and its potential impact on arthropod population dynamics. We compared herbivore-induced volatile emissions from Bacillus thuringiensis Berliner (Bt) maize plants to those from a nontransformed isoline following exposure to various types of leaf damage. When equal numbers of Helicoverpa zea Boddie (Lepidoptera: Noctuidae) larvae fed on Bt and non-Bt maize, volatile emissions were significantly lower in the transgenic plants, which also exhibited less leaf damage. When damage levels were controlled by adding more larvae to Bt plants, the plants' volatile emissions increased but displayed significant differences from those of nontransgenic plants. Significantly higher amounts of linalool, beta-myrcene, and geranyl acetate were released from transgenic maize than from non-Bt plants. Manipulating the duration of feeding by individual larvae to produce similar damage patterns resulted in similar volatile profiles for Bt and non-Bt plants. Controlling damage levels more precisely by mechanically wounding leaves and applying larval regurgitant likewise resulted in similar emission patterns for Bt and non-Bt maize. Overall, changes in the herbivore-induced volatile profiles of Bt maize appeared to be a consequence of altered larval feeding behavior rather than of changes in biochemical plant defense pathways. The implications of these findings for understanding the impacts of plant-mediated cues on pest and natural enemy behavior in transgenic crop systems are discussed. PMID- 16718568 TI - Volatile emissions from an odorous plant in response to herbivory and methyl jasmonate exposure. AB - Induced volatile terpenes have been commonly reported among diverse agricultural plant species, but less commonly investigated in odorous plant species. Odorous plants synthesize and constitutively store relatively large amounts of volatiles, and these may play a role in defense against herbivores. We examined the effect of herbivory and methyl jasmonate (MeJA) exposure on the release of volatile organic compounds (VOCs) in the marsh elder, Iva frutescens, which contains numerous constitutive VOCs, mainly mono- and sesquiterpenes. Our specific goal was to test for the presence of inducible VOCs in a naturally occurring plant already armed with VOCs. The abundant, native specialist leaf beetle Paria aterrima was used in herbivore induction trials. VOCs were sampled from herbivore wounded and unwounded, and from MeJA treated and untreated I. frutescens. Total VOC emissions were significantly greater in response to herbivory and MeJA treatment compared to unwounded controls. Herbivore wounding caused a substantial shift in the emission profile (42 VOCs from wounded, compared to 8 VOCs from unwounded I. frutescens), and MeJA had a similar yet less substantial influence on the emission pattern (28 VOCs from MeJA treated compared to 8 VOCs from untreated I. frutescens). Constitutive VOC emissions predominated, but some VOCs were detected only in response to herbivory and MeJA treatment, suggesting de novo synthesis. Several VOCs exhibited a delayed emission profile in contrast to the rapid release of constitutive VOCs, and principal components analysis revealed they were not associated with constitutive emissions. While I. frutescens contains many constitutive VOCs that are released immediately in response to herbivory, it also produces novel VOCs in response to feeding by the specialist P. aterrima and MeJA treatment. PMID- 16718569 TI - Isothiocyanates stimulating oviposition by the diamondback moth, Plutella xylostella. AB - Recognition of cabbage as a host plant for the diamondback moth (DBM) has previously been shown to depend on compounds that are extracted by soaking intact foliage in chloroform. Analysis of such chloroform extracts by open column chromatography has now resulted in the isolation of highly active fractions that elicit oviposition on treated filter papers. Further separation of these fractions by high-performance liquid chromatography revealed the presence of two distinct groups of active compounds that may be classified as volatile and non volatile. The two prominent volatile components were separated and identified by mass spectrometry as the isothiocyanates, iberin (3-methylsulfinylpropyl isothiocyanate) and sulforaphane (4-methylsulfinyl-3-butenyl isothiocyanate). Subsequent bioassays of a range of isothiocyanates showed that iberin and sulforaphane were the most active of those tested. Other isothiocyanates with sulfur in the side chain were also active, whereas alkyl and phenyl isothiocyanates had only limited activity. In electrophysiological experiments, electroantennograms (EAGs) indicated positive responses of moth antennae to the isothiocyanates that were most active in behavioral assays. Since sulforaphane has been identified as a major inducer of anticarcinogenic activity in mouse tissue, a synthetic analog (exo-2-acetyl-5-isothiocyanatonorbornane) that shows similar inducer activity was tested on DBM. This bicyclic analog was highly active in both behavioral and EAG assays, suggesting similarity in receptor sites for the two types of biological activity. PMID- 16718570 TI - Defensive secretion components of the host Parastizopus armaticeps as kairomones for the cleptoparasite Eremostibes opacus. AB - The subsocial tenebrionid Parastizopus armaticeps Per. is parasitized by the closely related Eremostibes opacus Koch (Coleoptera: Tenebrionidae). We found that the pygidial defensive secretions of both species are similar and contain a mixture of 1,4-benzoquinones, 1-alkenes, and monoterpene hydrocarbons. The 1 alkenes are dominated by 1-undecene, with admixtures of 1-tridecene in both species and 1-pentadecene in P. armaticeps only. Methyl- and ethyl-1,4 benzoquinone are the major quinones of the secretions of both species. The monoterpene fractions consist of (-)-alpha-pinene, (-)-camphene, sabinene, (-) beta-pinene, and (-)-limonene. Volatiles trapped with Porapak Q at the entrance to the breeding burrows of P. armaticeps were identified as components of the defensive secretion. However, in contrast to the secretion, the 1,4-benzoquinones were almost completely absent in the volatiles. Bioassays investigating attraction showed that the cleptoparasite E. opacus was drawn to the monoterpene hydrocarbons, produced by P. armaticeps, and deterred by the 1,4-benzoquinones. The 1-alkenes had no effect. Among the monoterpenes, only (-)-camphene was attractive to E. opacus. This is one of the rare cases of chemical exploitation of defensive allomones, and the first based on odor homology. We have drawn an evolutionary scenario including various functional changes in the defensive secretion compounds, leading to the kairomonal exploitation. PMID- 16718571 TI - Geographic and seasonal variation in alkaloid-based chemical defenses of Dendrobates pumilio from Bocas del Toro, Panama. AB - Poison frogs contain an alkaloid-based chemical defense that is derived from a diet of certain alkaloid-containing arthropods, which include mites, ants, beetles, and millipedes. Variation in population-level alkaloid profiles among species has been documented, and more than 800 different alkaloids have been identified. In the present study, we examine individual alkaloid variation in the dendrobatid poison frog Dendrobates pumilio among seven populations and between two seasons on Isla Bastimentos, located in the Bocas del Toro archipelago of Panama. Alkaloid profiles vary among populations and between seasons, illustrating that chemical defense in this species can vary on a small spatial and temporal scale. Alkaloid variation among populations is marginally correlated with geographic distance, and close populations have profiles more similar to each other than to distant populations. Individuals within populations also vary in alkaloid profiles. Differences are attributed to both spatial and temporal variations in the availability of alkaloid-containing arthropods. Many of the alkaloids present in the skin of D. pumilio appear likely to be of ant origin, supporting the importance of myrmecophagy in chemical defense among poison frogs. However, a variety of frog skin alkaloids was recently detected in mites, suggesting that mites may also play an important role in chemical defense. PMID- 16718572 TI - Quantitative GC-ECD analysis of halogenated metabolites: determination of surface and within-thallus elatol of Laurencia obtusa. AB - Information on natural concentrations or variability of secondary metabolites in marine organisms may be important both to ecological/evolutionary and applied approaches. A gas chromatographic procedure with an electron capture detector (GC ECD) was developed to quantify the sesquiterpenoid elatol at the surface and within-thalli of 70 specimens of the red seaweed Laurencia obtusa. The concentration of elatol was highest within-thalli [9.89 mg g(-1) of L. obtusa, dry weight (d.w.)], compared to lower values found at the surface [0.006 mg g(-1) of L.obtusa (d.w.), or 0.5-10.0 ng cm(-2)]. This method provides a rapid and inexpensive quantification of small quantities of elatol, and probably may also be used to quantify other halogenated compounds usually found in red seaweeds. PMID- 16718573 TI - Comparison of glass vessels and plastic bags for enclosing living plant parts for headspace analysis. AB - Plants release volatile chemicals into their surrounding air space that can affect the physiology of neighboring plants and influence the behavior of insects. In studying these interactions, it is desirable to collect volatiles from plants that have not been excised and are growing under as natural conditions as possible. We compared a vessel of borosilicate glass and Nylon-6 or polyester [poly(ethyleneterephthalate) or PET] cooking bags for enclosing plants during collection of volatiles. A push-pull airflow system was used, and volatiles were trapped on Tenax TA and analyzed by gas chromatography after thermal desorption. Low levels of impurities were found for the glass vessel and polyester bags. Nylon bags contained higher levels and more impurities. Recoveries of standards of 10 plant volatiles were measured in static and dynamic systems. In a static air system, there was good recovery only from the glass vessel. In a dynamic system, there was generally good recovery from both the glass vessel and polyester bags. Recoveries of alpha-pinene and (Z)-jasmone were poor throughout. The former was shown to have a very low breakthrough volume on the Tenax TA adsorbent, and the latter may be strongly adsorbed on glass. All three materials were essentially transparent in the IR and visible (photosynthetic) range but with significantly different absorptions in the UV range. In a simulated dynamic entrainment in full sunlight, internal vessel temperatures were higher than ambient by up to 9.5 degrees C in the glass vessel and 7.5 degrees C in the polyester bag. Lower increases in temperature relative to ambient (<1 degrees C) were recorded when entrainments were conducted in the shade. In a field trial, the profiles of volatiles collected from an apple tree infested with rosy apple aphid using a glass vessel and a polyester bag were similar. Polyester bags are recommended as more convenient than glass vessels for the enclosure of plants during the collection of volatiles. PMID- 16718574 TI - Identification of sex pheromone components of a New Zealand geometrid moth, the common forest looper Pseudocoremia suavis, reveals a possible species complex. AB - Gas chromatography-electroantennographic detection analysis of sex pheromone gland extracts of the common forest looper Pseudocoremia suavis (Lepidoptera: Geometridae), a polyphagous defoliator of introduced Pinaceae and many New Zealand trees, revealed four compounds that elicited antennal responses. The two major active compounds (6Z)-cis-9,10-epoxynonadec-6-ene and (3Z,6Z)-cis-9,10 epoxynonadeca-3,6-diene were identified by comparison with known standards. Of the two minor active compounds, one was tentatively identified as (3Z,6Z)-cis 9,10-epoxyhenicosa-3,6-diene, whereas the other could not be identified because of insufficient amounts in extracts. (6Z)-cis-9,10-Epoxynonadec-6-ene, (3Z,6Z) cis-9,10-epoxynonadeca-3,6-diene, and (3Z,6Z)-cis-9,10-epoxyhenicosa-3,6-diene were present in P. suavis gland extracts from Eyrewell Forest, a Pinus radiata plantation in the South Island of New Zealand, in a ratio of 35:65:5, respectively. Trapping trials in Eyrewell Forest established that (6Z)-cis-9,10 epoxynonadec-6-ene attracted male P. suavis. However, addition of (3Z,6Z)-cis 9,10-epoxyhenicosa-3,6-diene to the lure at <10% of (6Z)-cis-9,10-epoxynonadec-6 ene reduced capture of male moths, suggesting that one of its enantiomers was acting as a behavioral antagonist. During January-March of 2005, a blend trial involving single, binary, and ternary mixtures of the three components at Eyrewell Forest and at three other sites (two in the South Island and one in the North Island) revealed the existence of a second taxon of P. suavis at the three additional sites that was attracted to lures containing (3Z,6Z)-cis-9,10 epoxynonadeca-3,6-diene, either singly or in binary and ternary mixtures with (6Z)-cis-9,10-epoxynonadec-6-ene and (3Z,6Z)-cis-9,10-epoxyhenicosa-3,6-diene. This second taxon was not attracted to lures loaded solely with (6Z)-cis-9,10 epoxynonadec-6-ene. PMID- 16718575 TI - Individual, sexual, seasonal, and temporal variation in the amount of sagebrush lizard scent marks. AB - Although many animals deposit scent marks, previous studies have focused almost entirely on rodents or on the chemical structure of the signal. Here, we study the quantity and temporal pattern of chemical deposition by the territorial sagebrush lizard Sceloporus graciosus, measuring both femoral pore and fecal deposits. Specifically, we tested whether variation in deposition is a good cue of individual and sexual identity and/or whether it is more closely associated with body size and reproductive state, indicators of physiological condition. The results support the latter hypothesis. We found that although the amount of fluid deposited on a single perch (rarely quantified in mammals) carries little information on individual or sexual identity, it reflects the physiological condition and reproductive state of individual lizards and is replenished on a roughly weekly cycle, potentially providing additional information on the producer's activity level. The amount of deposition may thus provide important information to chemical receivers making mate choice and territorial defense decisions. The results further suggest that seasonal increases in gland production allow lizards to mark more sites rather than to influence the quality of the signal on a single perch. PMID- 16718576 TI - Identification of phytotoxic substances from early growth of barnyard grass (Echinochloa crusgalli) root exudates. AB - Barnyard grass is a problematic weed worldwide. It competes with crops and causes reduction in crop yields. In this study, barnyard grass suppressed rice emergence, and the degree of rice inhibition was proportional to the density of barnyard grass. Root exudates of barnyard grass reduced germination and growth of lettuce, rice, and monochoria. Fifteen compounds potentially involved in the phytotoxic activities of barnyard grass were isolated and identified, including phenolics, long-chain fatty acids, lactones, diethyl phthalate, acenaphthene, and derivatives of phthalic acids, benzoic acid, and decane. Quantities of diethyl phthalate, decanoic acid, myristic acid, stearic acid, 7,8-dihydro-5,6 dehydrokavain, and 7,8-dihydrokavain were 2.7, 11.1, 19.6, 35.5, 10.3, and 15.5 microg/ml of barnyard grass root exudates, respectively. The two lactones exhibited the greatest inhibition, followed by the phenolics and the derivatives of phthalic acids. Fatty acids had stronger suppression than diethyl phthalate and ethyl ester-4-ethoxy-benzoic acid. The acenaphthene and decane derivatives were the least phytotoxic. The phytotoxins released by barnyard grass roots showed strong inhibition on growth of broadleaf indicator plants and paddy weeds, but were less effective on barnyard grass itself and rice. Our study revealed that in addition to competition, barnyard grass also interferes with rice and other plants in its surroundings by chemical means. PMID- 16718577 TI - The court of public opinion: lay perceptions of polygraph testing. AB - We sampled a total of 411 participants and randomly assigned them one of three brief trial vignettes that contained either no-polygraph evidence, evidence of a passed polygraph test, or evidence of a failed polygraph test. Participants rendered guilt judgments and answered a series of questions concerning the trial in particular, and polygraph tests in general. Similar to previous studies on the impact of polygraph evidence on jurors' guilt judgments, this sample of jury eligible adults indicated that they did not find polygraph test evidence to be persuasive. Moreover, it mattered little to participants whether the results indicated the defendant passed a polygraph test, or that he failed a polygraph test. However, when our findings are compared to those of previous surveys involving experts in the field of psychophysiology, they differ in a number of important respects. The implications for decisions regarding admissibility (e.g., U.S. v. Alexander, 1975 and U.S. v. Scheffer, 1998) are discussed. PMID- 16718578 TI - Overriding psychiatric advance directives: factors associated with psychiatrists' decisions to preempt patients' advance refusal of hospitalization and medication. AB - Psychiatric advance directives (PADs) are intended to support patients' treatment decisions during a crisis. However, PAD statutes give clinicians broad discretion over whether to carry out patients' advance instructions. This study uses data from a survey of psychiatrists (N=164) to examine reasons for overriding PADs. In response to a hypothetical vignette, 47% of psychiatrists indicated that they would override a valid, competently-executed PAD that refused hospitalization and medication. PAD override was more likely among psychiatrists who worked in hospital emergency departments; those who were concerned about patients' violence risk and lack of insight; and those who were legally defensive. PAD override was less likely among participants who believed that involuntary treatment is largely unnecessary in a high-quality mental health system. PMID- 16718581 TI - An empirical test of the behaviour analysis interview. AB - The present experiment is the first empirical test of the Behaviour Analysis Interview (BAI), an interview technique developed by F. E. Inbau, J. E. Reid, J. P. Buckley, & B. C. Jayne (2001) designed to evoke different verbal and non verbal responses from liars and truth-tellers. Inbau et al. expect liars to be less helpful than truth-tellers in investigations and to exhibit more nervous behaviours. Just the opposite predictions, however, follow from the deception literature, which notes that liars take their credibility less for granted and are therefore more aware of their responses and their impact on others. This suggests that liars' answers should be more helpful than truth-tellers' answers, and liars' non-verbal responses should appear more relaxed than truth-tellers' non-verbal responses. In the present experiment, 40 participants (undergraduate students) lied or told the truth about an event during a BAI interview. The interviews were coded according to Inbau et al.'s guidelines. The results showed that, compared to liars, truth-tellers (a) were more naive and evasive when explaining the purpose of the interview, and (b) were less likely to name someone who they felt certain did not commit the crime. Truth-tellers also exhibited more nervous behaviours. The results were consistent with the predictions of the deception literature, and directly opposed to the predictions of BAI. PMID- 16718582 TI - Delayed prosecutions of historic child sexual abuse: analyses of 2064 Canadian criminal complaints. AB - Recently, in many English-speaking countries, legal principles that had the effect of barring delayed criminal prosecutions have been abrogated. In these jurisdictions, criminal prosecutions of child sexual abuse that is alleged to have occurred in the distant past (historic child sexual abuse or HCSA) are a growing legal challenge. These cases raise myriad issues relevant to research and the development of public policy that would benefit from a considered exchange of ideas that is informed by a clear understanding of the phenomenon. Based on 2064 judicial decisions of Canadian criminal complaints of HCSA we describe the trial, the complainant, the accused, and the offence. In the context of these legal cases, we raise some of the germane issues as well as suggestions for future research and discussion that we believe are particularly current and pressing. PMID- 16718584 TI - Effects of lexical prosody and word familiarity on lexical access of spoken Japanese words. AB - Lexical prosody (e.g., stress and pitch accent) has been shown to constrain lexical activation of spoken words in various languages. In the present study, whether or not the constraint of lexical prosody is affected by word familiarity in lexical access of Japanese words was examined using a cross-modal priming task. The stimuli were pairs of prosodically different homophones (minimal accent pairs). When the targets were more familiar members of minimal accent pairs, the responses were facilitated by prior presentations of primes that were prosodically different homophones of the targets, suggesting that lexical prosody did not constrain lexical activation. In contrast, when less familiar members of minimal accent pairs were used as the targets, the prosodically different homophones did not facilitate the responses to the targets. These results suggest that the constraint of lexical prosody is not so strong but is affected by the factor of word relative familiarity. PMID- 16718585 TI - Metalexical awareness: development, methodology or written language? A cross linguistic comparison. AB - This study explores the ability to access word boundaries of pre-school children, using an on-line methodology (Karmiloff-Smith, Grant, Sims, Jones, & Cockle (1996). Cognition, 58, 197-219.), which has hardly been used outside English speaking countries. In a cross-linguistic study in the Netherlands and Norway, four and five-year-old children were asked to repeat the last word every time a narrator stopped reading a story. In total 32 target-words were used, both closed and open class words, and both monosyllabic and disyllabic words. The outcomes in both countries were different from those of the original English study (Karmiloff Smith et al., 1996): four- and five-year-olds were successful in only about 26% of the cases, whereas the success rate in the former English experiment was 75% for the younger and 96% for the older children. No differences were found between age groups and between open and closed class words. This methodology does reveal the ability to access word boundaries, but probably not because of the ease of the on-line methodology in itself, but rather because literacy introduces new representations of language, even in on-line processing. The outcomes implicate that the ability to mark word boundaries does not seem to be a valid indication of who is ready for reading. PMID- 16718583 TI - Role of parenting and maltreatment histories in unipolar and bipolar mood disorders: mediation by cognitive vulnerability to depression. AB - In this article, we review empirical research on the role of individuals' parenting and maltreatment histories as developmental antecedents for symptoms and diagnosable episodes of unipolar and bipolar spectrum disorders. Our review is focused on the following three overarching questions: (1) Do negative parenting and a history of maltreatment contribute risk to symptoms or diagnosable episodes of unipolar and bipolar disorders? (2) Are the associations of negative parenting and maltreatment histories with bipolar disorders similar to those for unipolar depression? and (3) Are the associations between negative parenting and maltreatment histories and unipolar and bipolar symptoms or disorders mediated by cognitive vulnerability to depression? We begin by discussing the methodological requirements for demonstrating a psychosocial risk factor and the methodological issues that plague the parenting and maltreatment literatures. Next, we review the extant studies on the role of parenting histories in unipolar and bipolar disorders. We consider the specificity and possible moderators of the parenting-mood disorder relationship, as well as cognitive vulnerability to depression as a mediator of this relationship. Then, we review studies on the association of maltreatment histories with unipolar and bipolar disorders and the role of cognitive vulnerability to depression as a mediator of this association. We conclude with an assessment of the state of the parenting and maltreatment literatures in unipolar and bipolar disorder with regard to our guiding questions. PMID- 16718586 TI - Prediction of molecular alignment of nucleic acids in aligned media. AB - We demonstrate--using the data base of all deposited DNA and RNA structures aligned in Pf1-medium and RDC refined--that for nucleic acids in a Pf1-medium the electrostatic alignment tensor can be predicted reliably and accurately via a simple and fast calculation based on the gyration tensor spanned out by the phosphodiester atoms. The rhombicity is well predicted over its full range from 0 to 0.66, while the alignment tensor orientation is predicted correctly for rhombicities up to ca. 0.4, for larger rhombicities it appears to deviate somewhat more than expected based on structural noise and measurement error. This simple analytical approach is based on the Debye-Huckel approximation for the electrostatic interaction potential, valid at distances sufficiently far away from a poly-ionic charged surface, a condition naturally enforced when the charge of alignment medium and solute are of equal sign, as for nucleic acids in a Pf1 phage medium. For the usual salt strengths and nucleic acid sizes, the Debye Huckel screening length is smaller than the nucleic acid size, but large enough for the collective of Debye-Huckel spheres to encompass the whole molecule. The molecular alignment is then purely electrostatic, but it's functional form is under these conditions similar to that for steric alignment. The proposed analytical expression allows for very fast calculation of the alignment tensor and hence RDCs from the conformation of the nucleic acid molecule. This information provides opportunities for improved structure determination of nucleic acids, including better assessment of dynamics in (multi-domain) nucleic acids and the possibility to incorporate alignment tensor prediction from shape directly into the structure calculation process. The procedures are incorporated into MATLAB scripts, which are available on request. PMID- 16718587 TI - Backbone 1H, 13C, and 15N resonance assignments of the N-terminal domain of FKBP38 (FKBP38NTD). PMID- 16718588 TI - Real-time computation of a patient's respiratory effort during ventilation. AB - OBJECTIVE: In this paper, a new algorithm is proposed to compute the spontaneously generated respiratory effort during ventilation. METHODS: The algorithm computes a ventilated patient's respiratory effort in real-time by analyzing the respiratory pressure and flow signals that are acquired from the ventilator. The method requires an initial period where the patient's respiratory muscles are fully relaxed, for example during or shortly after surgery. During this period the patient's inspiratory airway resistance R(in), the expiratory airway resistance R(ex), the lung-thorax compliance C(lt) and the residual pressure after an infinitely long expiration P(0) are estimated by fitting the measured flow onto the measured pressure at the mouth using a model of the patient's respiratory system. When the patient starts breathing, the relation between the measured pressure and the flow changes, from which the respiratory effort of the patient P(mus) can be computed. RESULTS: The pressure P(mus) can be computed in real-time by using an equivalent model of the respiratory system of the patient. The estimation can be done with a recursive least squares (RLS) method. Further, the resulting P(mus) signal appears to have a constant shape, in which the main changing factor is the maximum amplitude per breath. CONCLUSION: The respiratory effort increases over time until the patient is disconnected from the ventilator. We hope the maximum amplitude can be used as an indicator of the pressure the muscles of the patient are able to produce. This amplitude of the (mus)-signal in combination with the standard deviation (SD) may eventually lead to a new indicator to determine the moment that the patient can be weaned from the ventilator. This will have to be examined in the future. PMID- 16718589 TI - Ligophorus pilengas Sarabeev & Balbuena, 2004 (Monogenea: Ancyrocephalidae) is proposed as the senior synonym of L. gussevi Miroshnichenko & Maltsev, 2004. AB - Ligophorus pilengas Sarabeev & Balbuena, 2004 and L. gussevi Miroshnichenko & Maltsev, 2004, both recently described as new species from the so-iuy mullet Mugil soiuy in the Sea of Azov and Black Sea, were compared. A morphometric analysis of both forms strongly suggests that they represent the same species. According to the 'Principle of priority', L. pilengas is proposed as a senior synonym of L. gussevi. PMID- 16718590 TI - Descriptions of Wardula bartolii n. sp. (Digenea: Mesometridae) and three newly recorded accidental parasites of Boops boops L. (Sparidae) in the NE Atlantic. AB - Four species of digeneans parasitic in the bogue Boops boops from the Spanish coast of the NE Atlantic are described. All are new records for this host, but one species is new and the other three are considered to be accidental parasites. Wardula bartolii n. sp. (Mesometridae) is distinguished from its two congeners on the basis of a range of morphometrical features, such as the post-ovarian extent of the uterus, the relative length of the attachment organ, the position of the pharynx and vitelline fields, and the absence of anterior caecal diverticula. The other three species are Lecithaster confusus Odhner, 1905, Aponurus laguncula Looss, 1907 (Lecithasteridae) and Accacladium serpentulum Odhner, 1928) (Accacoeliidae). PMID- 16718591 TI - Catatropis hatcheri n. sp. (Digenea: Notocotylidae) from Heleobia hatcheri (Prosobranchia: Hydrobiidae) and notes on its life-cycle in Patagonia, Argentina. AB - A new species of Catatropis Oghner, 1905 from a freshwater Neotropical prosobranch snail, Heleobia hatcheri (Hydrobiidae), is described. Naturally infected snails were collected from Nahuel Huapi Lake in Andean Patagonia. The characteristics of the larval stages are also presented. Experimental adults were recovered from the distal region of the intestinal caeca of chicks and ducklings and natural adults from a wild duck Anas platyrhynchos. Adults of Catatropis hatcheri n. sp. can be distinguished from all other species of the genus in having 10-12 (11) ventral glands in each lateral row, the cirrus-sac extending back to between the first third and the middle of the body, the metraterm shorter than the cirrus-sac, a previtelline field of 1,258-1,544 (1,396), vitelline follicles reach back to the anterior border of the testes with some follicles extending slightly lateral to them, only external testicular margin lobed and genital pore in median line just posterior to the intestinal bifurcation. In addition, the eggs have one filament on each pole, the rediae contain one or two mature cercariae, and the cercariae are tri-oculate, with a long tail and encyst in the environment. PMID- 16718592 TI - Megalophallus deblocki n. sp. (Digenea: Microphallidae) from Rostrhamus sociabilis (Vieillot) (Aves: Accipitridae) in Paraguay. AB - A new species in the little-known digenean fauna of Paraguayan birds is described in detail, especially in relation to its terminal genitalia. The new form, Megalophallus deblocki n. sp., is a microphallid from the intestine of the snail kite Rostrhamus sociabilis (Vieillot). It is differentiated in detail from its congeners, but differs mainly in having a substantially larger phallus (e.g. in relation to the ventral sucker), a smaller ventral sucker and smaller eggs. Comments are included on the functional morphology of the terminal genitalia. PMID- 16718593 TI - Diplectanids infesting the gills of the barramundi Lates calcarifer (Bloch) (Perciformes: Centropomidae), with the proposal of Laticola n. g. (Monogenoidea: Diplectanidae). AB - Four species of the Monogenoidea, Laticola lingaoensis n. sp., L. latesi (Tripathi, 1957) n. comb. [previously Pseudorhabdosynochus latesi (Tripathi, 1957) Kritsky & Beverley-Burton, 1986], L. paralatesi (Nagibina, 1976) n. comb. [previously Diplectanum paralatesi Nagibina, 1976] and Diplectanum penangi Liang & Leong, 1991, are reported from the gills of Lates calcarifer (Centropomidae) from the South China Sea (new geographical records for L. latesi and D. penangi). Collections from off Bathurst Island, Northern Territory, Australia, represent a new geographic record for L. paralatesi; Chilka Lake, Orissa, India, is established as the type-locality for L. latesi. Laticola n. g. (Diplectanidae) is proposed for species with a spoon-shaped copulatory organ with two to four concentric incomplete ridges in the base. Laticola lingaoensis, the type-species of Laticola, is described, and L. latesi and L. paralatesi are redescribed based on specimens from the South China Sea. Pseudorhabdosynochus monosquamodiscusi Balasuriya & Leong, 1995 and Pseudorhabdosynochus yangjiangenesis Wu & Li, 2005 are considered junior subjective synonyms of L. latesi and L. paralatesi, respectively. PMID- 16718594 TI - Selachohemecus benzi n. sp. (Digenea: Sanguinicolidae) from the blacktip shark Carcharhinus limbatus (Carcharhinidae) in the northern Gulf of Mexico. AB - Selachohemecus benzi Bullard & Overstreet n. sp. infects the heart and kidney of the blacktip shark Carcharhinus limbatus in the northern Gulf of Mexico off Florida and Mississippi, USA. Specimens of S. olsoni Short, 1954, the only congener and only other named blood fluke reported from a chondrichthyan in the Gulf of Mexico, were collected from the heart of the Atlantic sharpnose shark Rhizoprionodon terraenovae from two new localities, Apalachicola Bay, Florida, and Mississippi Sound, Mississippi, USA. The new species differs from S. olsoni by having a larger body (1.4-3.8 mm long), robust tegumental body spines numbering 51-63 along each lateral body margin, a testis extending from the posterior caeca to the ovary, and a medial ovary with lobes. We amend the diagnosis of Selachohemecus Short, 1954 to accommodate it and provide a diagnostic key for all named chondrichthyan blood flukes. PMID- 16718595 TI - Pseudorhabdosynochus venus n. sp. (Monogenea: Diplectanidae) from Epinephelus howlandi (Perciformes: Serranidae) off New Caledonia. AB - Pseudorhabdosynochus venus n. sp. is described from specimens collected from the gills of Epinephelus howlandi off Noumea, New Caledonia, South Pacific. The male sclerotised quadriloculate organ of P. venus, 61-101 microm in internal length, has an anterior chamber with a thin anterior wall, a very short cone and a short posterior tube. The sclerotised vagina, 50-67 microm in total length, is composed of an anterior open trumpet, an S-shaped canal, a tear-shaped principal chamber and a spherical accessory chamber; all parts are heavily sclerotised. The two squamodiscs have 10-11 rows of separate rodlets and no central closed row of rodlets. P. venus is differentiated from all other species of Pseudorhabdosynochus by the spectacular morphology of its sclerotised vagina. It is the first diplectanid described from E. howlandi. PMID- 16718596 TI - Microbial iron transport, storage and metabolism. Proceedings of a conference. June 1-4, 2005. Paris, France. PMID- 16718597 TI - Gene regulation by transmembrane signaling. AB - Studies of the ferric citrate transport genes in Escherichia coli K-12 have revealed a novel type of transcriptional regulation. The inducer, ferric citrate, binds to an outer membrane protein and must not be transported into the cells to initiate transcription of the ferric citrate transport genes. Rather, a signaling cascade from the cell surface across the outer membrane, the periplasm, and the cytoplasmic membrane into the cytoplasm transmits information on the presence of the inducer in the culture medium into the cytoplasm, where gene transcription occurs. The outer membrane protein FecA serves as a signal receiver and as a signal transmitter across the outer membrane. The FecR protein serves as a signal receiver in the periplasm and as a signal transmitter across the cytoplasmic membrane into the cytoplasm, where the FecI sigma factor is activated to bind RNA polymerase and specifically initiate transcription of the fecABCDE transport genes by binding to the promoter upstream of the fecA gene. Transcription of the fecI fecR regulatory genes is repressed by Fe2+ bound to the Fur repressor protein. Under iron-limiting conditions, Fur is not loaded with Fe2+, the fecI and fecR genes are transcribed, and the FecI and FecR proteins are synthesized and respond to the presence of ferric citrate in the medium when ferric citrate binds to the FecA protein. Regulation of the fec genes represents the paradigm of a growing number of gene regulation systems involving transmembrane signaling across three cellular compartments. PMID- 16718598 TI - Ternary complex formation facilitates a redox mechanism for iron release from a siderophore. AB - While the naturally occurring reducing agents glutathione (GSH) and ascorbate (H2A) alone are ineffective at reducing iron(III) sequestered by the siderophore ferrioxamine B, the addition of an iron(II) chelator, sulfonated bathophenanthroline (BPDS), facilitates reduction by either reducing agent. A mechanism is described in which a ternary complex is formed between ferrioxamine B and BPDS in a rapidly established pre-equilibrium step, which is followed by rate limiting reduction of the ternary complex by glutathione or ascorbate. Spectral, thermodynamic, and kinetic evidence are given for ternary complex formation. Ascorbate was found to be slightly more efficient at reducing the ternary complex than glutathione (k4=2.1 x 10(-3) M(-1) s(-1) and k4=6.3 x 10(-4) M(-1) s(-1), respectively) at pH 7. Reduction is followed by a rapid ligand exchange step where iron is released from ferrioxamine B to form tris (BPDS)iron(II). The implications of these results for siderophore mediated iron transport and release are discussed. PMID- 16718599 TI - Characterization of TonB interactions with the FepA cork domain and FecA N terminal signaling domain. AB - The mechanism of TonB dependent siderophore uptake through outer membrane transporters in Gram-negative bacteria is poorly understood. In an effort to expand our knowledge of the interaction between TonB and the outer membrane transporters, we have cloned and expressed the FepA cork domain (11-154) from Salmonella typhimurium and characterized its interaction with the periplasmic C terminal domain of TonB (103-239) by isotope assisted FTIR and NMR spectroscopy. For comparison we also performed similar experiments using the FecA N-terminal domain (1-96) from Escherichia coli which includes the conserved TonB box. The FepA cork domain was completely unfolded in solution, as observed for the E. coli cork domain previously [Usher et al. (2001) Proc Natl Acad Sci USA 98, 10676 10681]. The FepA cork domain was found to bind to TonB, eliciting essentially the same chemical shift changes in TonB C-terminal domain as was observed in the presence of TonB box peptides. The FecA construct did not cause this same structural change in TonB. The binding of the FepA cork domain to TonB-CTD was found to decrease the amount of ordered secondary structure in TonB-CTD. It is likely that the FecA N-terminal domain interferes with TonB-CTD binding to the TonB box. Binding of the FepA cork domain induces a loss of secondary structure in TonB, possibly exposing TonB surface area for additional intermolecular interactions such as potential homodimerization or additional interactions with the barrel of the outer membrane transporter. PMID- 16718600 TI - Feo--transport of ferrous iron into bacteria. AB - Bacteria commonly utilise a unique type of transporter, called Feo, to specifically acquire the ferrous (Fe2+) form of iron from their environment. Enterobacterial Feo systems are composed of three proteins: FeoA, a small, soluble SH3-domain protein probably located in the cytosol; FeoB, a large protein with a cytosolic N-terminal G-protein domain and a C-terminal integral inner membrane domain containing two 'Gate' motifs which likely functions as the Fe2+ permease; and FeoC, a small protein apparently functioning as an [Fe-S]-dependent transcriptional repressor. We provide a review of the current literature combined with a bioinformatic assessment of bacterial Feo systems showing how they exhibit common features, as well as differences in organisation and composition which probably reflect variations in mechanisms employed and function. PMID- 16718601 TI - Iron transport in Escherichia coli: all has not been said and done. AB - During recent years new systems involved in iron transport were identified in the old workhorse Escherichia coli (and in other enterobacteria). This came as a bit of a surprise because one might think transport of this essential trace element was already thoroughly studied. Moreover, it appears that iron homeostasis consists not only of uptake but also of efflux of this potentially toxic redox active metal. New findings in E. coli will be discussed and compared to the situation in other bacteria. PMID- 16718602 TI - Iron and pathogenesis of Shigella: iron acquisition in the intracellular environment. AB - Shigella species are able to grow in a variety of environments, including intracellularly in host epithelial cells. Shigella have a number of different iron transport systems that contribute to their ability to grow in these diverse environments. Siderophore iron uptake systems, heme transporters, and ferric and ferrous iron transport systems are present in these bacteria, and the genes encoding some of these systems appear to have spread among the Shigella species by horizontal transmission. Iron is not only essential for growth of Shigella but also plays an important role in regulation of metabolic processes and virulence determinants in Shigella. This regulation is mediated by the repressor protein Fur and the small RNA RyhB. PMID- 16718603 TI - Parasitism of iron-siderophore receptors of Escherichia coli by the siderophore peptide microcin E492m and its unmodified counterpart. AB - Microcin E492 (MccE492) is an antibacterial peptide naturally secreted by Klebsiella pneumoniae RYC492. Initially described as an 84-residue unmodified peptide, it was also recently isolated in a posttranslationally modified form, MccE492m. The production of MccE492m is dependent on the synthesis of enterobactin and the mceABCDEFGHIJ gene cluster. The posttranslational modification was characterized as a trimer of N-(2,3-dihydroxybenzoyl)-L-serine (DHBS) linked to the Ser84-carboxylate via a beta-D-glucose moiety. MccE492m was shown to bind ferric ions through the trimer of DHBS. This is the first example of a novel type of antibacterial peptide termed siderophore-peptide. Recognition of MccE492m, but also of the unmodified MccE492, was shown to be mediated by the catecholate siderophore receptors FepA, Cir and Fiu at the outer membrane of E. coli. The siderophore-type modification was shown to be responsible for a significant enhancement of the microcin antibacterial activity. Therefore, we propose that MccE492 and MccE492m use iron-siderophore receptors for uptake into the target bacteria and that improvement of MccE492 antimicrobial activity upon modification results from an increase in the microcin/receptor affinity. PMID- 16718604 TI - Iron acquisition and transport in Staphylococcus aureus. AB - Pathogenic Gram-positive bacteria encounter many obstacles in route to successful invasion and subversion of a mammalian host. As such, bacterial species have evolved clever ways to prevent the host from clearing an infection, including the production of specialized virulence systems aimed at counteracting host defenses or providing protection from host immune mechanisms. Positioned at the interface of bacteria/host interactions is the bacterial cell wall, a dynamic surface organelle that serves a multitude of functions, ranging from physiologic processes such as structural scaffold and barrier to osmotic lysis to pathogenic properties, for example the deposition of surface molecules and the secretion of cytotoxins. In order to succeed in a battle with host defenses, invading bacteria need to acquire the nutrient iron, which is sequestered within host tissues. A cell-wall based iron acquisition and import pathway was uncovered in Staphylococcus aureus. This pathway, termed the isd or iron-responsive surface determinant locus, consists of a membrane transporter, cell wall anchored heme binding proteins, heme/haptoglobin receptors, two heme oxygenases, and sortase B, a transpeptidase that anchors substrate proteins to the cell wall. Identification of the isd pathway provides an additional function to the already bountiful roles the cell wall plays in bacterial pathogenesis and provides new avenues for therapeutics to combat the rise of antimicrobial resistance in S. aureus. This review focuses on the molecular attributes of this locus, with emphasis placed on the mechanism of iron transport and the role of such a system during infection. PMID- 16718605 TI - Roles of environmental heme, and menaquinone, in streptococcus agalactiae. AB - Most bacteria require iron for growth. However, as it may not be directly available under aerobic conditions, bacteria may use iron-sequestering molecules, such as bacterially encoded siderophores, or heme, which is the major iron source in the animal host. Bacteria may also assimilate heme for purposes other than as an iron source. Once internalised, heme can activate, for example, a heme dependent catalase and a cytochrome oxidase. In bacterial pathogen Streptococcus agalactiae, heme, in association with exogenous menaquinone, activates a respiratory chain. Respiration has radical effects on carbon metabolism. GBS respiration-grown cells display improved survival in an aerobic environment and greater virulence in a murine septicemia model. GBS might benefit from its ecological niches to capture heme and menaquinone, i.e., from other bacteria when it colonizes host mucosa, or from blood-containing organs during septicemia. PMID- 16718606 TI - Neutrophil gelatinase-associated lipocalin, a siderophore-binding eukaryotic protein. AB - NGAL (neutrophil gelatinase-associated lipocalin) also known as lcn2 or siderochalin is constitutively expressed in myelocytes and stored in specific granules of neutrophils. It is highly induced in a variety of epithelial cells during inflammation. Analysis of the crystal structure of NGAL expressed in E.coli showed that NGAL has the ability to bind catecholate type siderophores and in this way prevent bacteria from acquisition of siderophore-bound iron. NGAL (or 24p3 as the highly homologous murine orthologue is named) knock out mice have a profound defect in defense against E.coli after intraperitoneal injection. This defect can be mimicked in wild-type mice by providing siderophore iron, which cannot be sequestered by NGAL, testifying to the specific role of NGAL as a siderophore binding protein in innate immunity. Megalin, a scavenger receptor functions as a receptor for NGAL and mediates uptake into endosomes, but other NGAL receptors are likely to exist. PMID- 16718609 TI - Planar relationships of the semicircular canals in two strains of mice. AB - The mouse is increasingly important as a subject of vestibular research. Although many studies have focused on the vestibular responses of mice to angular rotation, the geometry of their semicircular canals has not been described. High voltage X-ray computed tomography was used to measure the anatomy of the semicircular canals of two strains of mice, C57Bl/6J and CBA/CaJ. The horizontal plane of a stereotaxic coordinate system was defined by the midpoints of the external auditory meati and the point where the incisors emerge from the maxilla. The centroids of the lumens of the bony canals were calculated, and planes that describe the canals were fit using a least-squares regression analysis to the resulting points. Vectors normal to each regressed plane were used to represent the corresponding canal's axis of rotation, and angles of these vectors relative to skull landmarks as well as to each other were calculated. The horizontal canal of the mouse was found to be angled anteriorly upward 17.8 degrees for CBA/CaJ and 32.6 degrees for C57Bl/6J from the reference horizontal plane. Angles between ipsilateral canals deviated up to 12.3 degrees from orthogonal, and angles between contralateral synergistic canals (left anterior-right posterior, right anterior-left posterior, and horizontal-horizontal) deviated from parallel by up to 14.8 degrees. The orientations of the canals within the head as well as the orientations of the canals relative to each other were significantly different between the two strains, suggesting that care must be taken in the design and interpretation of developmental and physiologic studies involving different mouse strains. PMID- 16718611 TI - A novel TECTA mutation in a Dutch DFNA8/12 family confirms genotype-phenotype correlation. AB - A novel TECTA mutation, p.R1890C, was found in a Dutch family with nonsyndromic autosomal dominant sensorineural hearing impairment. In early life, presumably congenital, hearing impairment occurred in the midfrequency range, amounting to about 40 dB at 1 kHz. Speech recognition was good with all phoneme recognition scores exceeding 90%. An intact horizontal vestibuloocular reflex was found in four tested patients. The missense mutation is located in the zona pellucida (ZP) domain of alpha-tectorin. Mutations affecting the ZP domain of alpha-tectorin are significantly associated with midfrequency hearing impairment. Substitutions affecting other amino acid residues than cysteines show a significant association with hearing impairment without progression. Indeed, in the present family progression seemed to be absent. In addition, the presently identified mutation affecting the ZP domain resulted in a substantially lesser degree of hearing impairment than was previously reported for DFNA8/12 traits with mutations affecting the ZP domain of alpha-tectorin. PMID- 16718610 TI - Characterization of an abundant COL9A1 transcript in the cochlea with a novel 3' UTR: Expression studies and detection of miRNA target sequence. AB - EST N66408 represents one of several large unique clusters expressed in the Morton human fetal cochlear cDNA library. N66408 is 575 bp in size and initial BLAST analysis of this sequence showed no homology to any known genes or expressed sequence tags (ESTs) from other organs or tissues. Sequence of the original cochlear clone from which N66408 was derived revealed that the corresponding cDNA was about 700 bp in size, including 125 bp at its 5' end with homology to the 3' end of COL9A1 in addition to 575 bp of novel sequence. RT-PCR analysis using primers specific to COL9A1 isoforms 1 and 2 detected expression of both isoforms in human fetal cochlea. Tissue in situ hybridization using the novel 3' UTR sequence as probe showed abundant expression in spiral limbus and spiral ligament, and a moderate level of expression in the organ of Corti. dbEST analysis of ESTs specific to the 3' UTR of COL9A1 showed 19 ESTs derived from various tissues; three polyadenylation sites were identified and the majority of these ESTs were derived from overlapping polyadenylation signals at the second site (position 749-758). Comparison of the 3' UTR of human COL9A1 with its orthologs as well as with dbEST uncovered a highly conserved region around the overlapping polyadenylation signals at position 749-758 in mammals. A search of the microRNA database revealed a highly conserved target sequence for miR-9 immediately preceding the overlapping polyadenylation signals in the novel 3' UTR of COL9A1, suggesting its role in posttranscriptional regulation of COL9A1. PMID- 16718612 TI - Demonstration of a longitudinal concentration gradient along scala tympani by sequential sampling of perilymph from the cochlear apex. AB - Local applications of drugs to the inner ear are increasingly being used to treat patients' inner ear disorders. Knowledge of the pharmacokinetics of drugs in the inner ear fluids is essential for a scientific basis for such treatments. When auditory function is of primary interest, the drug's kinetics in scala tympani (ST) must be established. Measurement of drug levels in ST is technically difficult because of the known contamination of perilymph samples taken from the basal cochlear turn with cerebrospinal fluid (CSF). Recently, we reported a technique in which perilymph was sampled from the cochlear apex to minimize the influence of CSF contamination (J. Neurosci. Methods, doi: 10.1016/j.jneumeth.2005.10.008 ). This technique has now been extended by taking smaller fluid samples sequentially from the cochlear apex, which can be used to quantify drug gradients along ST. The sampling and analysis methods were evaluated using an ionic marker, trimethylphenylammonium (TMPA), that was applied to the round window membrane. After loading perilymph with TMPA, 10 1-muL samples were taken from the cochlear apex. The TMPA content of the samples was consistent with the first sample containing perilymph from apical regions and the fourth or fifth sample containing perilymph from the basal turn. TMPA concentration decreased in subsequent samples, as they increasingly contained CSF that had passed through ST. Sample concentration curves were interpreted quantitatively by simulation of the experiment with a finite element model and by an automated curve-fitting method by which the apical-basal gradient was estimated. The study demonstrates that sequential apical sampling provides drug gradient data for ST perilymph while avoiding the major distortions of sample composition associated with basal turn sampling. The method can be used for any substance for which a sensitive assay is available and is therefore of high relevance for the development of preclinical and clinical strategies for local drug delivery to the inner ear. PMID- 16718614 TI - A biophysical model of the inner hair cell: the contribution of potassium currents to peripheral auditory compression. AB - The term peripheral auditory compression refers to the fact that the whole range of audible sound pressure levels is mapped into a narrower range of auditory nerve responses. Peripheral compression is the by-product of independent compressive processes occurring at the level of the basilar membrane, the inner hair cell (IHC), and the auditory nerve synapse. Here, an electrical-circuit equivalent of an IHC is used to look into the compression contributed by the IHC. The model includes a mechanically driven transducer potassium (K(+)) conductance and two time- and voltage-dependent basolateral K(+) conductances: one with fast and one with slow kinetics. Special attention is paid to faithfully implement the activation kinetics of these basolateral conductances. Optimum model parameters are provided to account for previously reported in vitro observations that demonstrate the compression associated with the gating of the transducer and of the basolateral channels. Without having to readjust its parameters, the model also accounts for the in vivo nonlinear IHC transfer characteristics. Model simulations are then used to investigate the relative contribution of the transducer and basolateral K(+) currents to the nonlinear IHC input/output functions in vivo. The simulations suggest that the voltage-dependent activation of the basolateral currents compresses the DC potential for stereocilia displacements above approximately 5 nm. The degree of compression exceeds 2-to-1 and is similar for all stimulation frequencies. The AC potential is compressed in a similar way, but only for frequencies below 800 Hz. The simulations further suggest that the nonlinear gating of the transducer current is responsible for the expansive growth of the DC potential with increasing sound level (slope of 2 dB/dB) at low sound pressure levels. PMID- 16718616 TI - Nasal immunization with anthrax protective antigen protein adjuvanted with polyriboinosinic-polyribocytidylic acid induced strong mucosal and systemic immunities. AB - PURPOSE: The current anthrax vaccine adsorbed (AVA) was originally licensed for the prevention of cutaneous anthrax infection. It has many drawbacks, including the requirement for multiple injections and subsequent annual boosters. Thus, an easily administrable and efficacious anthrax vaccine is needed to prevent the most lethal form of anthrax infection, inhalation anthrax. We propose to develop a nasal anthrax vaccine using anthrax protective antigen (PA) protein as the antigen and synthetic double-stranded RNA in the form of polyriboinosinic polyribocytidylic acid (pI:C) as an adjuvant. METHODS: Mice were nasally immunized with recombinant PA admixed with pI:C. The resulting PA-specific antibody responses and the lethal toxin neutralization activity were measured. Moreover, the effect of pI:C on dendritic cells (DCs) was evaluated both in vivo and in vitro. RESULTS: Mice nasally immunized with rPA adjuvanted with pI:C developed strong systemic and mucosal anti-PA responses with lethal toxin neutralization activity. These immune responses compared favorably to that induced by nasal immunization with rPA adjuvanted with cholera toxin. Poly(I:C) enhanced the proportion of DCs in local draining lymph nodes and stimulated DC maturation. CONCLUSIONS: This pI:C-adjuvanted rPA vaccine has the potential to be developed into an efficacious nasal anthrax vaccine. PMID- 16718617 TI - A specific picomolar hybridization-based ELISA assay for the determination of phosphorothioate oligonucleotides in plasma and cellular matrices. AB - PURPOSE: To develop and validate an ultrasensitive and specific hybridization based enzyme-linked immunosorbent assay method for quantification of two phosphorothioate oligonucleotides (PS ODNs) (G3139 and GTI-2040) in biological fluids. METHODS: This assay was based on hybridization of analytes to the biotin labeled capture ODNs followed by ligation with digoxigenin-labeled detection ODN. The bound duplex was then detected by anti-digoxigenin-alkaline phosphatase using Attophos (Promega, Madison, WI, USA) as substrate. S1 nuclease and major factors such as the hybridization temperature, concentration of capture probe, and the use of detergent were evaluated toward assay sensitivity, selectivity, and accuracy. RESULTS: The method is selective to the parent drugs with minimal cross reactivity (<6%) with 3'-end deletion oligomers for both G3139 and GTI-2040. A linear range of 0.05 to 10 nM (r2 > 0.99) was observed for GTI-2040 in a variety of biological matrices. For both G3139 and GTI-2040, the within-day precision and accuracy values were found to be <20% and 90-110%, respectively; the between-day precision and accuracy were determined to be <20% and 90-120%. Addition of S1 nuclease combined with washing step greatly improved the assay linearity and selectivity. The utility of this assay was demonstrated by simultaneous determination of GTI-2040 in plasma and its intracellular levels in treated acute myeloid leukemia patients. CONCLUSIONS: The validated hybridization enzyme-linked immunosorbent assay method is specific for quantitation of PS ODNs in biological samples to picomolar level. This method provides a powerful technique to evaluate plasma pharmacokinetics and intracellular uptake of PS ODNs in patients and shows its utility in clinical evaluations. PMID- 16718620 TI - Effects of surface electrical stimulation both at rest and during swallowing in chronic pharyngeal Dysphagia. AB - We tested two hypotheses using surface electrical stimulation in chronic pharyngeal dysphagia: that stimulation (1) lowered the hyoid bone and/or larynx when applied at rest, and (2) increased aspiration, penetration, or pharyngeal pooling during swallowing. Bipolar surface electrodes were placed on the skin overlying the submandibular and laryngeal regions. Maximum tolerated levels of stimulation were applied while patients held their mouth closed at rest. Videofluoroscopic recordings were used to measure hyoid movements in the superior inferior and anterior-posterior dimensions and the subglottic air column position while stimulation was on or off. Patients swallowed 5 ml liquid when stimulation was off, at low sensory stimulation levels, and at maximum tolerated levels (motor). Speech pathologists, blinded to condition, tallied the frequency of aspiration, penetration, pooling, and esophageal entry from videofluorographic recordings of swallows. Only significant (p = 0.0175) hyoid depression occurred during stimulation at rest. Aspiration and pooling were significantly reduced only with low sensory threshold levels of stimulation (p = 0.025) and not during maximum levels of surface electrical stimulation. Those patients who had reduced aspiration and penetration during swallowing with stimulation had greater hyoid depression during stimulation at rest (p = 0.006). Stimulation may have acted to resist patients' hyoid elevation during swallowing. PMID- 16718618 TI - Zebra mussel antifouling activity of the marine natural product aaptamine and analogs. AB - Several aaptamine derivatives were selected as potential zebra mussel (Dreissena polymorpha) antifoulants because of the noteworthy absence of fouling observed on Aaptos sponges. Sponges of the genus Aaptos collected in Manado, Indonesia consistently produce aaptamine-type alkaloids. To date, aaptamine and its derivatives have not been carefully evaluated for their antifoulant properties. Structure-activity relationship studies were conducted using several aaptamine derivatives in a zebra mussel antifouling assay. From these data, three analogs have shown significant antifouling activity against zebra mussel attachment. Aaptamine, isoaaptamine, and the demethylated aaptamine compounds used in the zebra mussel assay produced EC(50) values of 24.2, 11.6, and 18.6 microM, respectively. In addition, neither aaptamine nor isoaaptamine produced a phytotoxic response (as high as 300 microM) toward a nontarget organism, Lemna pausicostata, in a 7-day exposure. The use of these aaptamine derivatives from Aaptos sp. as potential environmentally benign antifouling alternatives to metal based paints and preservatives is significant, not only as a possible control of fouling organisms, but also to highlight the ecological importance of these and similar biochemical defenses. PMID- 16718621 TI - Cine magnetic resonance imaging with single-shot fast spin echo for evaluation of dysphagia and aspiration. AB - The aim of this study was to determine the feasibility of and interest in evaluation of swallowing using dynamic magnetic resonance imaging (cine-MRI) in patients with dysphagia and aspiration caused by an abnormal pharyngeal phase of swallow. A cohort of six patients previously treated for head and neck cancer with persistent dysphagia and/or aspiration were evaluated an average of 47 months after treatment. The morphology and mobility of the oral, oropharyngeal, and laryngeal structures were analyzed using cine-MRI using single-shot fast spin echo technology. The qualitative observations were compared with a clinical fiberoptic swallowing evaluation. Swallowing physiology was analyzable for dry (saliva) swallow in all patients. MRI was well-tolerated by all six patients and no clinical aspiration occurred during the MRI. In five of six cases, further information on the cause of dysphagia was obtained using cine-MRI compared with the clinical evaluation alone. In the remaining case, cine-MRI confirmed the clinical evaluation. Cine-MRI using the dry swallow technique is feasible and without risk in patients with clinical aspiration. Cine-MRI is complementary to clinical evaluation of swallowing in patients with an abnormal pharyngeal phase of swallowing resulting from treatment of cancer. PMID- 16718622 TI - Is the anatomical protrusion on the posterior hypopharyngeal wall associated with cadavers of only the elderly? AB - Cricopharyngeal bars are commonly seen on a barium swallow radiologic examination and represent the failure of the cricopharyngeus to relax. Traditionally, the bars have been considered as functional or physiologic protrusions. Recently, anatomical cricopharyngeal protrusion has been found in about 30% of cadavers of the elderly, suggesting that such a structural change may become a physical barrier that affects the normal deglutition in a living person. This suggests that such a radiographic finding should be carefully considered when interpreting radiologic and manometrical examinations and managing dysphagia of the elderly. However, the finding of the anatomical cricopharyngeal protrusion was based on the observation of cadavers of the elderly (mean age = 77 years). The aim of this study was to further investigate whether such an anatomical cricopharyngeal protrusion exists in cadavers of other ages. Using the dissection method, we examined 63 human cadavers that were divided into three age groups: young adult (6 females and 10 males, age = 16-24 years old), adult (8 females and 23 males, age = 25-64 years old), and early elderly (5 females and 11 males, age = 65-69 years old). We found that 57 of the 63 cadavers had a smooth mucosal surface on the posterior hypopharyngeal and upper esophageal wall. A slightly thickened posterior muscular wall was observed at the cricoid level in six cadavers (five from the adult group and one from the early elderly group), but its boundary was not clearly identified. Taking the previous reports together, our results suggest that an anatomical cricopharyngeal protrusion is closely associated with the aging process. PMID- 16718623 TI - Laterality of swallowing in healthy subjects by AP projection using videofluoroscopy. AB - Although anteroposterior projections in videofluoroscopic examination of swallowing (VF) provide clinically important information, the laterality of swallowing in healthy subjects has not been examined fully to date. One hundred sixty-seven healthy volunteers were prospectively studied. The subjects were asked to swallow 5 ml of barium solution three times while X-ray images were taken that showed the pathway of the solution from the pharynx into the esophagus to assess the laterality of swallowing. We classified patterns of swallowing into three types according to passage in the pharyngoesophageal segment as indicated by width: RD (right-side-dominant flow), LD (left-side-dominant flow), and NL (no laterality in flow). Fifty-eight percent of the subjects were classified as NL, 35% as LD, and 7% as RD. The ratio of types in women was NL:LD:RD = 7:2:1. There were no significant differences among the groups according to age. Although classifications in young men (age = 20-30 years) tended to be the same as in women regardless of age, ratios of LD tended to increase with age; 71% of elderly adults (age = 51-75 years) were classified as LD. These results demonstrate laterality in normal swallowing and will be helpful in determining treatment strategies for the patients with dysphagia. PMID- 16718615 TI - CYP induction-mediated drug interactions: in vitro assessment and clinical implications. AB - Cytochrome P450 (CYP) induction-mediated interaction is one of the major concerns in clinical practice and for the pharmaceutical industry. There are two major issues associated with CYP induction: a reduction in therapeutic efficacy of comedications and an induction in reactive metabolite-induced toxicity. Because CYP induction is a metabolic liability in drug therapy, it is highly desirable to develop new drug candidates that are not potent CYP inducer to avoid the potential of CYP induction-mediated drug interactions. For this reason, today, many drug companies routinely include the assessment of CYP induction at the stage of drug discovery as part of the selection processes of new drug candidates for further clinical development. The purpose of this article is to review the molecular mechanisms of CYP induction and the clinical implications, including pharmacokinetic and pharmacodynamic consequences. In addition, factors that affect the degree of CYP induction and extrapolation of in vitro CYP induction data to in vivo situations will also be discussed. Finally, assessment of the potential of CYP induction at the drug discovery and development stage will be discussed. PMID- 16718624 TI - Leptin and prolactin, but not corticosterone, modulate body weight and thyroid function in protein-malnourished lactating rats. AB - To understand the role of hormonal changes in the lower food ingestion and body weight in protein-restricted lactating rats as well as the higher serum T (3), higher deiodination, iodide and T (3) milk transfer, we measured maternal serum prolactin, leptin, TSH and corticosterone, which are hormones that could influence those parameters. After birth, dams were separated into: control-fed with a 23 % protein diet (n = 12) and PR (protein-restricted)-fed with an 8 % protein diet (n = 12). At the 4 (th) and 21 (st) day of lactation, half of the animals in each group were sacrificed. PR dams presented hyperleptinemia (day 4: + 20 %; day 21: + 19 %; p < 0.05) and hypoprolactinemia (day 4: - 85 %; day 21: - 92 %; p < 0.05), which could help explain the lower food consumption and body weight in lactating PR rats since leptin is anorexigenic and prolactin is orexigenic. Also, this hyperleptinemia could contribute for the increase in serum T (3) of PR dams, since leptin stimulates T (3) production, especially acting on deiodinases. Serum corticosterone was not different between PR and C groups, and TSH was lower only at the end of lactation. Thus, we suggest that both leptin and prolactin could play an important role in the body weight and thyroid hormone changes observed in protein-malnourished lactating rats. PMID- 16718625 TI - Association between functional FABP2 promoter haplotype and type 2 diabetes. AB - Fatty acid-binding protein 2 (FABP2) is a cytosolic protein expressed exclusively in epithelial cells of the small intestine. Some, albeit not conclusive, evidence indicates that the Thr-allele of FABP2 Ala54Thr polymorphism is associated with type 2 diabetes. More recently, common FABP2 promoter polymorphisms have shown association with postprandial increase of triglycerides, body composition and plasma lipid levels. Therefore, we reasoned that variants in the FABP2 promoter may also predispose to type 2 diabetes mellitus. In our Caucasian study population, we found three SNPs and three insertion-deletion polymorphisms that are in complete linkage disequilibrium defining promoter haplotype A and B within 1kb 5' of the FABP2 initiation codon. Haplotype calculations indicated that the FABP2 promoter and Ala54Thr variants were strongly linked. Functional analysis of promoter fragments demonstrated that haplotype difference is caused by polymorphisms within 260 bp downstream of the FABP2 initiation codon. Using a prospective case-control study nested within the EPIC-Potsdam cohort of 192 incident type 2 diabetes cases and 384 sex-/age-matched controls, male subjects carrying the FABP2 haplotype B allele showed significantly decreased risk of type 2 diabetes when adjusted for BMI (OR = 0.50, 95 % CI = 0.28 - 0.87, p < 0.05) and additional covariates (OR = 0.42, 95 % CI 0.22 - 0.81, p < 0.01). Further adjustment for the Ala54Thr polymorphism revealed an OR of 0.18 (95 % CI 0.06 - 0.49, p < 0.001). Similarly, Ala/Ala homozygote males carrying the promoter haplotype B had decreased risk (0.33, 0.11 - 0.94, p < 0.05) of type 2 diabetes after stratification for the Ala54Thr polymorphism. FABP2 promoter haplotypes or genotype combinations defined by the promoter and Ala54Thr polymorphism were not associated with BMI, body fat, leptin, HbA (1c), total cholesterol or HDL. In conclusion, our findings suggest that the functional FABP2 promoter haplotype may contribute to type 2 diabetes in a sex-specific manner. PMID- 16718626 TI - Hyperthyroid hearts display a phenotype of cardioprotection against ischemic stress: a possible involvement of heat shock protein 70. AB - Hyperthyroid hearts are shown to display a phenotype of cardioprotection against ischemic stress, but the underlying signaling mechanisms remain largely unknown. The present study investigated the possible relation of HSP70 to the thyroid hormone induced cardioprotection. HSP70 is a redox-regulated molecular chaperone, and enhances cell survival under stress. Thyroxin (25 microg/100 g body weight) was administered to Wistar male rats for four days (THYR-4d) and two weeks (THYR 14d), respectively, while untreated animals served as controls (CON-4d, CON-14d). Isolated hearts from control and thyroxin treated rats were subjected to 20 min zero-flow ischemia followed by 45 min of reperfusion (I/R). The amount of HSP70 in the myocardium for THYR-14d was 1.85 times the levels of CON-14d (p < 0.05). The levels of HSP70 expression were no different between THYR-4d and CON-4d, p > 0.05. This was only accompanied by an increase in MDA levels (used as an index of oxidative stress) in THYR-14d compared to untreated hearts. These changes corresponded to a differential response of the heart to I/R; post-ischemic recovery of function was significantly increased in THYR-14d compared to CON-14d, and was no different between the THYR-4d and CON-4d hearts. In conclusion, long term thyroxin administration results in increased tolerance of the myocardium to I/R and enhances the expression of HSP70 which may, at least in part, account for this response. PMID- 16718627 TI - Intracerebroventricular injection of proinsulin C-peptide does not influence food consumption in male Long-Evans rats. AB - Insulin and C-peptide are released in equimolecular concentrations in the circulation after food ingestion as they result from the enzymatic cleavage of proinsulin. In the brain, insulin inhibits food intake through hypothalamic receptors. In the present study, we tested the ability of C-peptide to modulate food intake when injected in the brain lateral ventricle of Long-Evans rats. For this purpose, 10 adult male rats (BW 320 - 350 g) were deprived of food overnight. They were intra-cerebroventricularly injected with 10 microg C-pepitde or vehicle (artificial cerebrospinal fluid [CSF]) during the first hour of the light period and chow intake was measured 1, 3, 6 and 24 hours after injection. Chow availability immediately triggered food consumption. Food intake was not different between CSF- and C-peptide-injected rats either after one hour (5.7 +/- 0.6 g [CSF] vs. 6.7 g +/- 0.5 g; ns) or after 24 hrs (23.3 +/- 1.4 g [CSF] vs. 25.1 g +/- 1.4 g; ns). In addition, a higher dose (20 microg/rat) had no effect at all in satiated rats one hour after injection or later contrary to the 100 % increase measured after injection of 2 microg of neuropeptide Y. Thus, we conclude that contrary to insulin, C-peptide does not regulate feeding behaviour in normal rats whatever their insulin status. PMID- 16718628 TI - Meal energy content is related to features of meal-related ghrelin profiles across a typical day of eating in non-obese premenopausal women. AB - Some features of the meal-related profile of ghrelin correspond to acute energy intake, suggesting a role in short-term energy homeostasis. Yet, no studies have examined this relationship across a typical day of eating when effects of time of day and or cumulative energy intake may also exist. AIM OF STUDY: To examine the relation between ghrelin and acute energy intake by quantifying changes in ghrelin over 24 hours in response to three typical meals and a snack occurring throughout the day. METHODS: Fourteen non-obese women consumed three meals and a snack at specific times; total ghrelin was measured repeatedly over 24 hours. RESULTS: Significant correlations existed between: 1) meal calories and the post meal trough (r = - 0.36; p < 0.05), and 2) the sum of breakfast and lunch calories and subsequent dinner rise (r = - 0.45; p < 0.02) and 3) the sum of breakfast and lunch calories and dinner peak (r = - 0.54; p < 0.003). Thus, as energy intake increased across the day, pre-meal rises of subsequent meals were reduced. CONCLUSION: Meal-related profiles of ghrelin are not only associated with the energy content of specific meals, but also with the accumulated calories prior to a subsequent meal, suggesting a role in the modulation of acute energy homeostasis. PMID- 16718629 TI - The effect of oral glucose loads on tissue metabolism during angiotensin II receptor and beta-receptor blockade in obese hypertensive subjects. AB - AT1 receptor blockers and ACE inhibitors decrease the risk for new onset diabetes mellitus. The phenomenon could be related to a direct angiotensin II effect on tissue metabolism. To address the issue, we recruited eighteen obese hypertensive patients. Patients were randomized to double-blind treatment with either valsartan (n = 8) or atenolol (n = 10) for thirteen weeks. They underwent an oral glucose tolerance test before and during active treatment, while metabolism was monitored through subcutaneous and intramuscular microdialysis and indirect calorimetry. After glucose ingestion, venous glucose and insulin concentrations increased rapidly while systemic free fatty acid concentrations were suppressed. Dialysate glucose and lactate concentrations increased briskly in adipose tissue and in skeletal muscle. Dialysate glycerol decreased profoundly in both tissues. Respiratory quotient increased markedly after glucose ingestion. These responses were identical at baseline and during active treatment either drug. We conclude that AT1 receptor blockade in obese hypertensive patients has no effect on interstitial glucose supply, lipolysis, and substrate oxidation. One possible explanation is that angiotensin II levels in obese hypertensives are not sufficient to elicit the metabolic changes that have been observed after direct angiotensin II application. The exact mechanism by which inhibition of the renin angiotensin-aldosterone system decreases the diabetes risk remains unresolved and requires further study. PMID- 16718630 TI - Circulating progesterone and obesity in men. AB - Progesterone can be detected in male plasma and has been considered to originate mainly from the adrenals. We have examined the association between circulating progesterone and obesity in a sample of thirty-eight lean to morbidly obese men aged 44.5 +/- 9.9 years (BMI: 44.3 +/- 12.8 kg/m (2)). Plasma concentrations of progesterone, 17-OH-progesterone as well as androstenedione, testosterone, DHT and DHEA-S were determined. Negative correlations were observed between plasma progesterone levels and body weight (r = - 0.47, p < 0.05), BMI (r = - 0.56, p < 0.001), waist circumference (r = - 0.58, p < 0.001), as well as subcutaneous adipocyte diameter (r = - 0.50, p < 0.05). Plasma levels of 17-OH-progesterone, DHEA-S, androstenedione, testosterone and DHT were also negatively associated with body weight, BMI and waist circumference. However, the ratio of 17-OH progesterone-to-progesterone and androstenedione-to-17-OH-progesterone were not related to these variables. A positive correlation was found between circulating progesterone and DHEA-S levels (r = 0.50, p < 0.002 after adjustment for age). Accordingly, using multivariate regression analyses, the best steroid predictor of progesterone level was plasma DHEA-S. Waist circumference was the best predictor of progesterone levels in a multivariate model including steroid concentrations as well as waist circumference, BMI and subcutaneous adipocyte diameter. In conclusion, plasma progesterone was negatively associated with markers of obesity such as BMI, waist circumference and subcutaneous adipocyte diameter in this sample of men. Circulating DHEA-S level was the best steroid correlate of plasma progesterone. We suggest that the low progesterone levels observed in obese men may reflect decreased adrenal C(19) steroid production in the adrenal cortex. Further research is needed to confirm this hypothesis. PMID- 16718631 TI - Association of serum adiponectin concentration to lipid and glucose metabolism in healthy humans. AB - BACKGROUND: Adiponectin is a recently discovered plasma protein with many associations to glucose and lipid metabolism. Due to its central role in cardiovascular diseases and insulin resistance, we studied the relationship between serum adiponectin and factors reflecting glucose and lipid metabolism. METHODS AND RESULTS: Thirty healthy participants (20M/10F, age 32.0 +/- 2.1 years, BMI 25.8 +/- 0.9 kg/m (2) and HbA (1c) 5.2 +/- 0.1 %) were studied four times at approximately one week intervals. The effects of a 4-hour euglycemic hyperinsulinemia (40 mU/m (2)/min), saline infusion (control), oral glucose, and oral fat load on serum adiponectin were studied. No significant correlation was found between serum adiponectin and insulin sensitivity before (r = 0.25) or after adjustment for age, BMI and gender (r = 0.04). Adiponectin concentration correlated inversely with HbA (1c) (r = - 0.43, p < 0.05), insulin concentration (r = - 0.38, p < 0.05) and triglyceride concentration (r = - 0.42, p < 0.05) but positively with HDL cholesterol (r = 0.38, p < 0.05). Metabolic procedures had no effect on serum adiponectin. CONCLUSIONS: Our findings favor the interpretation that adiponectin is not causally related to insulin sensitivity in healthy participants. The strongest associations of adiponectin in healthy participants are to be found to lipid metabolism. Serum levels of adiponectin are very stable and not acutely affected by hyperinsulinemia, oral glucose or fat load. PMID- 16718632 TI - Preliminary evidence of FABP2 A54T polymorphism associated with reduced risk of type 2 diabetes and obesity in women from a German cohort. AB - The T54 variant of the FABP2 gene has shown an association with the insulin resistance syndrome in some, but not all, studies. Here, we tested the hypothesis that the association between FABP2 A54T genotype and type 2 diabetes (T2DM) is confounded by body mass index (BMI) and is different between the two genders. 192 incident cases of T2DM and 384 sex- and age-matched controls were taken from the EPIC-Potsdam study cohort. Logistic regression analyses revealed that BMI was a strong confounder for diabetes risk association among women. When adjusted for BMI, the homozygous T54 variant was significantly associated with reduced risk of T2DM in women (OR = 0.24, 95 %CI: 0.07 - 0.82), but not in men in the co-dominant inheritance model. Accordingly, HbA (1c) values were significantly lower in women carrying two T54 alleles with BMI regarded as covariate. While accounting for potentially confounding effects, linear trends of increased BMI and leptin values were observed in women according to the presence of T54 alleles. The interaction term (p = 0.04) of continuous BMI and T54-coding genotypes suggested that the T54 variant is an effect-modifier for BMI in females. We conclude that the T54 allele of FABP2 A54T is associated both with higher BMI and reduced risk of T2DM in women from the German EPIC-Potsdam study. PMID- 16718633 TI - Increased TNF-alpha and decreased TGF-beta expression in peripheral blood leukocytes after acute myocardial infarction. AB - Inflammation contributes to the development of atherosclerosis and cardiovascular events. Counteracting pro- and anti-inflammatory responses of serum cytokines have been reported, but the relevance of TNF-alpha, TGF-beta and IL-6 gene expression in peripheral blood leukocytes and their contribution to systemic inflammation in atherosclerosis, especially after acute myocardial infarction (AMI), has not been investigated yet. Using quantitative RT-PCR, we determined temporal cytokine mRNA expression alterations in blood cells from patients with AMI (n = 51). Serum cytokine concentrations were analyzed in parallel using the ELISA technique. TNF-alpha mRNA expression rates and serum concentrations were significantly elevated in AMI patients compared to controls (n = 77), while mRNA expression and serum content of TGF-beta were decreased. Interestingly, we found no statistically significant correlation between transcript and protein levels, indicating that gene expression in leukocytes may be an independent sign for systemic inflammation. While IL-6 was significantly increased in serum from AMI patients with positive correlation to left ventricular dysfunction and negative correlation to ejection fraction, IL-6 mRNA levels did not differ between patients and controls. Gene expression alterations indicate a sophisticated regulation of counteracting TNF-alpha and TGF-beta cytokine expression in peripheral blood leukocytes after AMI with bias towards a pro-inflammatory situation. PMID- 16718634 TI - Evaluation of the components of the insulin-like growth factors system in GH deficient adults: effects of twelve-month rhGH treatment. AB - The impact of GH deficiency and rhGH replacement therapy on IGF-I, IGFBP-3 and ALS levels has been widely studied. There is less information available on IGF-II levels, the component of the ternary complex poorly dependent on GH. We investigate the components of IGFs system in 36 GHD adults (28M, 8F, age 45 +/- 14 yrs) before and after 12 months of rhGH therapy (mean dose 0.3 +/- 0.1 mg/day). One-hundred healthy sex- and age-matched subjects were studied for comparison. At baseline, GHD patients showed IGF-I and IGF-II levels and IGFs to IGFBP-3 molar ratios that were lower than controls. During therapy, IGF-I levels increased (p < 0.01) to normal range. IGF-II levels, though higher than at baseline (p < 0.01), remained lower than in controls (p < 0.01). ALS and IGFBP-3 significantly increased (p < 0.001). These modifications resulted in normalization in IGF-I to IGFBP-3 ratio, while no change in IGF-II to IGFBP-3 ratio was observed. In conclusion, the increase of serum IGF-II levels during rhGH treatment in GHD patients probably reflects the increase in the other components of ternary complex (ALS and IGFBP-3). However, serum IGF-II levels as well as IGF-II to IGFBP-3 ratio, although increased, were definitely lower than in controls. This last result, given the increasing evidences of a direct implication of IGF-II in cancer, may further confirm the safety of rhGH replacement in adults with severe GHD as diagnosed by appropriate stimulation tests. PMID- 16718635 TI - Cannabinoid receptor signaling directly inhibits thermogenesis and alters expression of adiponectin and visfatin. PMID- 16718636 TI - Pre-diabetes and metabolic syndrome in Germans. PMID- 16718637 TI - Determination of polycyclic aromatic hydrocarbons in olive oil by a completely automated headspace technique coupled to gas chromatography-mass spectrometry. AB - A new and completely automated method for the determination of ten relevant polycyclic aromatic hydrocarbons (PAHs) in olive oil is proposed using an extraction by the headspace (HS) technique. Quantification and confirmation steps are carried out by gas chromatography-mass spectrometry (GC-MS) combining simultaneously selected-ion monitoring (SIM) and tandem mass spectrometry (MS/MS). This combination offers on one hand an increased sensitivity and on the other hand, selective and reliable qualitative information. Sample pretreatment or clean-up are not necessary because the olive oil sample is put directly into an HS vial, automatically processed by HS and introduced into the GC-MS instrument for analysis. Because of its high selectivity and sensitivity, a triple-quadrupole (QqQ) detector coupled with the gas chromatograph allows us to limit handling. Each sample is completely processed in approximately 63 min (45 min for HS isolation and 18 min for GC-MS determination), a reduced time compared with previously published methods. The chemical and instrumental variables were preliminarily optimized using uncontaminated olive oil samples spiked with 25 microg kg(-1) of each target compound. The final method was validated to ensure the quality of the results. The precision was satisfactory, with relative standard deviation (RSD) values in the range 3-9%. Recovery rates ranged from 96 to 99%. Limits of detection (LOD) were calculated as 0.02-0.06 microg kg(-1) and the limits of quantification (LOQ) were obtained as 0.07-0.26 microg kg(-1). It must be mentioned that the LOD and LOQ are much lower than the maximum levels established by the European Union (EU) in oils and fats intended for direct human consumption or for use as an ingredient in foods, which are set at 2 microg kg( 1). All the figures of merit are completely in accordance with the latest EU legislation. This fact makes it possible to consider the proposed method as a useful tool for the control of PAHs in olive oils. PMID- 16718638 TI - Optimizing UV laser focus profiles for improved MALDI performance. AB - Matrix assisted laser desorption/ionization (MALDI) applications, such as proteomics, genomics, clinical profiling and MALDI imaging, have created a growing demand for faster instrumentation. Since the commonly used nitrogen lasers have throughput and life span limitations, diode-pumped solid-state lasers are an alternative. Unfortunately this type of laser shows clear performance limitations in MALDI in terms of sensitivity, resolution and ease of use, for applications such as thin-layer sample preparations, acceptance of various matrices (e.g. DHB for glycopeptides) and MALDI imaging. While it is obvious that the MALDI process has some dependence on the characteristics of the laser used, it is unclear which features are the most critical in determining laser performance for MALDI. In this paper we show, for the first time, that a spatially structured laser beam profile in lieu of a Gaussian profile is of striking importance. This result enabled us to design diode-pumped Nd : YAG lasers that on various critical applications perform as well for MALDI as the nitrogen lasers and in some respects even better. The modulation of the beam profile appears to be a new parameter for optimizing the MALDI process. In addition, the results trigger new questions directing us to a better understanding of the MALDI process. PMID- 16718639 TI - Determination of the propylene oxide-hemoglobin adduct by gas chromatography electron impact ionization mass spectrometry. AB - A gas chromatography-mass spectrometry method was developed for the determination of the propylene oxide (PO)-hemoglobin adduct. The adduct was identified as hydroxy propyl valine (HPV), and released from globin by the modified Edman degradation and extracted with ethyl ether. HPV and deuterated HPV (d6-HPV) were synthesized for identification and quality control. d6-HPV was used as an internal reference standard. The dried extract was completely derivatized with N methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA). The method detection limit (MDL) of the assay was 10 pmole/g for HPV, based on assayed hemoglobin of 0.1 g. The method was applied to the determination of the PO-hemoglobin adduct formed in young female Sprague-Dawley rats after treatment for 4 and 5 weeks with 5 and 10 mM PO via drinking water. HPV was detected by the proposed procedure. After 4 weeks, the concentration of HPV was 6.75 nmole/g hemoglobin during treatment with 5 mM, and 80.26 nmole/g hemoglobin during treatment with 10 mM. The adduct level in 5 weeks increased up to about 51.47 nmole/g during treatment with 5 mM PO in the drinking water and up to about 120.27 nmole/g during treatment with 10 mM PO. This method was also applied to determine the concentrations of HPV in the blood of 20 persons living near the Ulsan petroleum industrial complex in Korea. As a result, HPV-hemoglobin adduct was detected in the concentration range 0-1100 pmol/g in the human blood samples. PMID- 16718640 TI - Desorption/ionization on self-assembled monolayer surfaces (DIAMS). PMID- 16718641 TI - Serum iohexol analysis by micellar electrokinetic capillary chromatography. AB - A simple and rapid ( approximately 4 min) method for the measurement of iohexol in serum for assessing the glomerular filtration rate is described. It is based on direct serum injection on the capillary by MEKC. The method is linear between 8 and 260 mg/L, with an RSD of peak height of 2.9%. Several simple steps have contributed to an improved daily precision, such as choosing a high pH buffer, increasing the SDS concentration, frequent standardization, and eliminating any sample pretreatment. PMID- 16718643 TI - Capillary electrophoresis for the analysis of glycoprotein pharmaceuticals. AB - Carbohydrate chains in glycoprotein pharmaceuticals play important roles for the expression of their biological activities, but the structure and compositions of carbohydrate chains are dependent on the conditions for their production. Therefore, evaluation of the carbohydrate chains is quite important for productive process development, characterization of product for approval application, and routine quality control. The oligosaccharides themselves have complex structure including blanching and various glycosidic linkages, and oligosaccharides in one glycoprotein pharmaceutical generally have high heterogeneity, and characterization of oligosaccharide moiety in glycoprotein has been a challenging target. In these situations, CE has been realized as a powerful tool for oligosaccharide analysis due to its high resolution and automatic operating system. This review focuses on the application of CE to the glycoform analysis of glycoproteins and profiling of the N-linked glycans released from glycoprotein pharmaceuticals. Current applications for structure analysis using CE-MS(n) technique and glycan profiling method for therapeutic antibody are also described. PMID- 16718642 TI - Interlaboratory study of a NACE method for the determination of R-timolol content in S-timolol maleate: assessment of uncertainty. AB - Analyses of statistical variance were applied to evaluate the precision and practicality of a CD-based NACE assay for R-timolol after enantiomeric separation of R- and S-timolol. Data were collected in an interlaboratory study by 11 participating laboratories located in Europe and North America. General qualitative method performance was examined using suitability descriptors (i.e. resolution, selectivity, migration times and S/N), while precision was determined by quantification of variances in the determination of R-timolol at four different impurity levels in S-timolol maleate samples. The interlaboratory trials were designed in accordance with the ISO guideline 5725-2. This allowed estimating for each sample, the different variances, i.e. between-laboratory (s2(Laboratories)), between-day (s2(Days)) and between-replicate (s2(Replicates)). The variances of repeatability (s2r) and reproducibility (s2R) were then calculated. The estimated uncertainty, derived from the precision estimates, seems to be concentration-dependent above a given threshold. This example of R-timolol illustrates how a laboratory can evaluate uncertainty in general. PMID- 16718644 TI - Micellar electrokinetic capillary chromatographic method for the quantitative analysis of uricosuric and antigout drugs in pharmaceutical preparations. AB - A simple MEKC method is described for the separation and quantification of seven widely used uricosuric and antigout drugs, including allopurinol (AP), benzbromarone (BZB), colchicine (COL), orotic acid (OA), oxypurinol (OP), probenecid (PB), and sulfinpyrazone (SPZ). The drugs were separated in a BGE of borate buffer (45 mM; pH 9.00) with SDS (20 mM) as the micellar source and the separated drugs were directly monitored with a UV detector (214 nm). Several parameters affecting the separation and analysis of the drugs were studied. Based on the normalized peak-area ratios of the drugs to an internal standard versus the concentration of the drugs, the method is applicable to quantify BZB, COL, and SPZ (each 5-200 microM), AP, OA, OP, and PB (each 10-200 microM) with detection limits (S/N = 3, 0.5 psi, 5 s injection) in the range of 0.6-4.0 microM. The precision (RSD; n = 5) and accuracy (relative error; n = 5) of the method for intraday and interday analyses of the analytes at three levels (30, 120, and 180 microM) are below 4% (n = 3). The method was demonstrated to be suitable for the analysis of AP and COL in commercial tablets with speed and simplicity. PMID- 16718645 TI - A novel Bicine running buffer system for doubled sodium dodecyl sulfate - polyacrylamide gel electrophoresis of membrane proteins. AB - A novel, Bicine-based SDS-PAGE buffer system was developed for the analysis of membrane proteins. The method involves molecular weight-based separations of fully denatured and solubilized proteins in two dimensions. This doubled SDS-PAGE (dSDS-PAGE) approach produced a diagonal arrangement of protein spots and successfully circumvented problems associated with membrane proteome analysis involving traditional gel-based methods. Membrane proteins from the anaerobic bacterium Clostridium thermocellum were used for these investigations. Tricine dSDS-PAGE and the newly developed Bicine-dSDS-PAGE were compared with the standard glycine-dSDS-PAGE (Laemmli protocol) in their suitability to separate C. thermocellum membrane proteins. Large-format gel experiments using optimized gel preparation and running buffer conditions revealed a 112% increase in protein spot count for Tricine-dSDS-PAGE and a 151% increase for Bicine-dSDS-PAGE, compared to glycine-dSDS-PAGE. The data clearly indicated that Bicine-dSDS-PAGE is a superior method for the analysis of membrane proteins, providing enhanced resolution and protein representation. PMID- 16718646 TI - Development and validation of a capillary electrophoresis method with laser induced fluorescence detection for the determination of captopril in human urine and pharmaceutical preparations. AB - This study describes the development of a CE method for the analysis of the antihypertensive drug captopril using LIF detection. The method is based on the derivatization of captopril with the fluorescent label 5 iodoacetamidofluorescein. The optimization of the electrophoretic electrolyte composition together with other variables, such as applied voltage and injection time, resulted in a solution of 20 mM phosphate buffer adjusted to pH 12.0. The calibration curve for the fluorescent captopril derivative was linear in the concentration range 3.5-6000 ng/mL with a detection limit of 0.5 ng/mL. Intra- and interday precision (at a concentration of about 100 times the LOD) were less than 0.86 and 1.16%, respectively, both expressed as RSD. The assay was successfully used for quantification of captopril in some marketed pharmaceutical preparations and urine samples. PMID- 16718647 TI - Determination of phenothiazines in pharmaceutical formulations and human urine using capillary electrophoresis with chemiluminescence detection. AB - A CE instrument coupled with chemiluminescence (CL) detection was designed for the determination of promethazine hydrochloride (PTH) and promazine hydrochloride (PMH) in real samples. An important enhancement of the CL emission of luminol with potassium ferricyanide was observed in the presence of these phenothiazines; so this system was selected for their detection after CE separation. Parameters affecting the electrophoretic separation were optimized in a univariate way, while those affecting CL detection were optimized by means of a multivariate approach based on the use of experimental designs. Chemometrics was also employed for the study of the robustness of the factors influencing the postcolumn CL detection. The method allows the separation of the phenothiazines in less than 4 min, achieving LODs of 80 ng/mL for PMH and 334 ng/mL for PTH, using sample injection by gravity. Electrokinetic injection was used to obtain lower LODs for the determination of the compounds in biological samples. The applicability of the CE-CL method was illustrated in the determination of PTH in pharmaceutical formulations and in the analysis of PMH in human urine, using a previous SPE procedure, achieving an LOD of 1 ng/mL and recoveries higher than 85%. PMID- 16718648 TI - Capillary electrochromatography with zwitterionic stationary phase on the lysine bonded poly(glycidyl methacrylate-co-ethylene dimethacrylate) monolithic capillary column. AB - A polymer-based neutral monolithic capillary column was prepared by radical polymerization of glycidyl methacrylate and ethylene dimethacrylate in a 100 mum id fused-silica capillary, and the prepared monolithic column was subsequently modified based on a ring opening reaction of epoxide groups with 1 M lysine in solution (pH 8.0) at 75 degrees C for 10 h to produce a lysine chemically bonded stationary phases in capillary column. The ring opening reaction conditions were optimized so that the column could generate substantial EOF. Due to the zwitterionic functional groups of the lysine covalently bonded on the polymer monolithic rod, the prepared column can generate cathodic and anodic EOF by varying the pH values of running buffer during CEC separation. EOF reached the maximum of -2.0 x 10(-8) m2v(-1)s(-1) and 2.6 x 10(-8) m2v(-1)s(-1) with pH of the running buffer of 2.25 and 10, respectively. As a consequence, neutral compounds, ionic solutes such as phenols, aromatic acids, anilines, and basic pharmaceuticals were all successfully separated on the column by CEC. Hydrophobic interaction is responsible for separation of neutral analytes. In addition, the electrostatic and hydrophobic interaction and the electrophoretic migration play a significant role in separation of the ionic or ionizable analytes. PMID- 16718649 TI - Capillary electrophoresis contributions to the hydromorphone metabolism in man. AB - CE-ESI multistage IT-MS (CE-MS(n), n < or = 4) and computer simulation of fragmentation are demonstrated to be effective tools to detect and identify phase I and phase II metabolites of hydromorphone (HMOR) in human urine. Using the same CE conditions as previously developed for the analysis of urinary oxycodone and its metabolites, HMOR and its phase I metabolites produced by N-demethylation, 6 keto-reduction and N-oxidation and phase II conjugates of HMOR and its metabolites formed with glucuronic acid, glucose, and sulfuric acid could be detected in urine samples of a patient that were collected during a pharmacotherapy episode with daily ingestion of 48 mg of HMOR chloride. The CE MS(n) data obtained with the HMOR standard, synthesized hydromorphol and hydromorphone-N-oxide, and CYP3A4 in vitro produced norhydromorphone were employed to identify the metabolites. This approach led to the identification of previously unknown HMOR metabolites, including HMOR-3O-glucide and various N oxides, structures for which no standard compounds or mass spectra library data were available. Furthermore, the separation of alpha- and beta-hydromorphol, the stereoisomers of 6-keto-reduced HMOR, was achieved by CE in the presence of the single isomer heptakis(2,3-diacetyl-6-sulfato)-beta-CD. The obtained data indicate that the urinary excretion of alpha-hydromorphol is larger than that of beta-hydromorphol. PMID- 16718650 TI - From oogonia to mature oocytes: inactivation of the maternal centrosome in humans. AB - The fine structure of human oogonia and growing oocytes has been reviewed in fetal and adult ovaries. Preovulatory maturation and the ultrastructure of stimulated oocytes from the germinal vesicle (GV) stage to metaphase II (MII) stage are also documented. Oogonia have large nuclei, scanty cytoplasm with complex mitochondria. During folliculogenesis, follicle cell processes establish desmosomes and deep gap junctions at the surface of growing oocytes, which are retracted during the final stages of maturation. The zona pellucida is secreted in secondary follicles. Growing oocytes have mitochondria, Golgi, rough endoplasmic reticulum (RER), ribosomes, lysosomes, and lipofuscin bodies, often associated with Balbiani bodies and have nuclei with reticulated nucleoli. Oocytes from antral follicles show numerous surface microvilli and cortical granules (CGs) separated from the oolemma by a band of microfilaments. The CGs are evidently secreted by Golgi membranes. The GV oocytes have peripheral Golgi complexes associated with a single layer of CGs close to the oolemma. They have many lysosomes, and nuclei with dense compact nucleoli. GV breakdown occurs by disorganization of the nuclear envelope and the oocyte enters a transient metaphase I followed by MII, when it is arrested and ovulated. Maturation of oocytes in vitro follows the same pattern of meiosis seen in preovulatory oocytes. The general organization of the human oocyte conforms to that of most other mammals but has some unique features. The MII oocyte has the basic cellular organelles such as mitochondria, smooth endoplasmic reticulum, microfilaments, and microtubules, while Golgi, RER, lysosomes, multivesicular, residual and lipofuscin bodies are very rare. It neither has yolk nor lipid inclusions. Its surface has few microvilli, and 1-3 layers of CGs, aligned beneath the oolemma. Special reference has been made to the reduction and inactivation of the maternal centrosome during oogenesis. The MII spindle, often oriented perpendicular to the oocyte surface, is barrel-shaped, anastral and lacks centrioles. Osmiophilic centrosomes are not demonstrable in human eggs, since the maternal centrosome is nonfunctional. However, oogonia and growing oocytes have typical centrioles, similar to those of somatic cells. The sperm centrosome activates the egg and organizes the sperm aster and mitotic spindles of the embryo, after fertilization. PMID- 16718651 TI - Neural basis of first and second language processing of sentence-level linguistic prosody. AB - A fundamental question in multilingualism is whether the neural substrates are shared or segregated for the two or more languages spoken by polyglots. This study employs functional MRI to investigate the neural substrates underlying the perception of two sentence-level prosodic phenomena that occur in both Mandarin Chinese (L1) and English (L2): sentence focus (sentence-initial vs. -final position of contrastive stress) and sentence type (declarative vs. interrogative modality). Late-onset, medium proficiency Chinese-English bilinguals were asked to selectively attend to either sentence focus or sentence type in paired three word sentences in both L1 and L2 and make speeded-response discrimination judgments. L1 and L2 elicited highly overlapping activations in frontal, temporal, and parietal lobes. Furthermore, region of interest analyses revealed that for both languages the sentence focus task elicited a leftward asymmetry in the supramarginal gyrus; both tasks elicited a rightward asymmetry in the mid portion of the middle frontal gyrus. A direct comparison between L1 and L2 did not show any difference in brain activation in the sentence type task. In the sentence focus task, however, greater activation for L2 than L1 occurred in the bilateral anterior insula and superior frontal sulcus. The sentence focus task also elicited a leftward asymmetry in the posterior middle temporal gyrus for L1 only. Differential activation patterns are attributed primarily to disparities between L1 and L2 in the phonetic manifestation of sentence focus. Such phonetic divergences lead to increased computational demands for processing L2. These findings support the view that L1 and L2 are mediated by a unitary neural system despite late age of acquisition, although additional neural resources may be required in task-specific circumstances for unequal bilinguals. PMID- 16718652 TI - Quantitative and reliable in vitro method combining scanning electron microscopy and image analysis for the screening of osteotropic modulators. AB - The increased generation and up-regulated activity of bone resorbing cells (osteoclasts) play a part in the impairment of bone remodeling in many bone diseases. Numerous drugs (bisphosphonates, calcitonin, selective estrogen receptor modulators) have been proposed to inhibit this increased osteoclastic activity. In this report, we describe a pit resorption assay quantified by scanning electron microscopy coupled with image analysis. Total rabbit bone cells with large numbers of osteoclasts were cultured on dentin slices. The whole surface of the dentin slice was scanned and both the number of resorption pits and the total resorbed surface area were measured. Resorption pits appeared at 48 h and increased gradually up to 96 h. Despite the observation of a strong correlation between the total resorption area and the number of pits, we suggest that area measurement is the most relevant marker for osteoclastic activity. Osteotropic factors stimulating or inhibiting osteoclastic activity were used to test the variations in resorption activity as measured with our method. This reproducible and sensitive quantitative method is a valuable tool for screening for osteoclastic inhibitors and, more generally, for investigating bone modulators. PMID- 16718653 TI - Analysis of intersubject variability in activation: an application to the incidental episodic retrieval during recognition test. AB - Since the brain may engage different neuronal systems for a single behavioral goal, activation may show intersubject variability expressed in the systematic recruitment of multiple distinct networks. We apply a principal component analysis (PCA) to activation over task conditions and subjects to reveal cortical networks that may underlie this intersubject variability. Normal subjects were presented with novel meaningless objects, which appeared in personally familiar or unfamiliar places. During a subsequent, event-related functional MRI (fMRI) experiment, each subject was presented with learned or novel objects in isolation and performed a learned/novel judgment. Recollection of places was not essential for the task, and may exhibit large intersubject variations. The right posterodorsal posterior cingulate cortex (pPCC) and left retrosplenial cortex, whose involvement in place-recognition has been previously established, were selected as regions of interest for the PCA. Neural responses to objects associated with familiar relative to unfamiliar places in pPCC, but not in the retrosplenial cortex, were negatively correlated with task-related activation (common over all objects) in the right anterolateral prefrontal cortex and the left intraparietal sulcus. The latter areas have been implicated previously in cognitive control and object recognition, respectively. These results suggest right prefrontal control over neural processes both in the left parietal cortex, related to object-recognition (enhancement), and pPCC, related to nonessential recollection of place-memory (suppression), but not in the retrosplenial cortex, related to the sense of familiarity. This analysis revealed an important aspect of functional anatomy that was not detectable using a conventional analysis of average activations. PMID- 16718654 TI - Volitional control of attention and brain activation in dual task performance. AB - This study used functional MRI (fMRI) to examine the neural effects of willfully allocating one's attention to one of two ongoing tasks. In a dual task paradigm, participants were instructed to focus either on auditory sentence comprehension, mental rotation, or both. One of the major findings is that the distribution of brain activation was amenable to strategic control, such that the amount of activation per task was systematically related to the attention-dividing instructions. The activation in language processing regions was lower when attending to mental rotation than when attending to the sentences, and the activation in visuospatial processing regions was lower when attending to sentences than when attending to mental rotations. Additionally, the activation was found to be underadditive, with the dual-task condition eliciting less activation than the sum of the attend sentence and attend rotation conditions. We also observed a laterality shift across conditions within language-processing regions, with the attend sentence condition showing bilateral activation, while the dual task condition showed a left hemispheric dominance. This shift suggests multiple language-processing modes and may explain the underadditivity in activation observed in the current and previous studies. PMID- 16718655 TI - Adult survivors of childhood cancer and unemployment: A metaanalysis. AB - A range of late effects is associated with the survival of childhood cancer, including problems with employment. The purpose of this metaanalysis was to assess the risk of unemployment of adult survivors of childhood cancer compared with healthy controls and to explore prognostic factors. A literature search of studies published between 1966 and January 2006 was conducted using the databases of MedLine, CINAHL, EMBASE, ClinPSYCH, PsycINFO, and OSHROM. The authors synthesized data using a random effects model. A total of 34 articles was found, in which 40 original empirical studies were reported, 24 of which were controlled studies. Survivors of childhood cancer were nearly twice as likely to be unemployed than healthy controls (odds ratio [OR] 1.85, 95% confidence interval [95% CI], 1.27-2.69). Survivors of central nervous system (CNS) and brain tumors were nearly 5 times more likely to be unemployed (OR 4.74, 95% CI, 1.21-18.65), whereas the risks for survivors of blood or bone cancers were elevated but not found to be statistically significant (OR 1.42, 95% CI, 0.79-2.55; OR 1.97, 95% CI, 0.88-4.40, respectively). No increased risk was found for survivors of other or mixed diagnoses (OR 0.97, 95% CI, 0.27-3.53). Furthermore, survivors in the U.S. had an overall 3-fold risk (OR 3.24, 95% CI, 2.16-4.86) of becoming unemployed, whereas no such risk was found for European survivors (OR 1.00, 95% CI, 0.58-1.70). Apart from type of diagnosis and country, predictors of unemployment were younger age, lower education or intelligence quotient, female gender, motor impairment or epilepsy, and radiotherapy. Adult survivors of childhood cancer are at risk of unemployment, especially the subgroup of survivors of CNS and brain tumors. Interventions to enhance participation in work life should be developed and evaluated. PMID- 16718656 TI - Mouse oocyte differentiation during antral follicle development. AB - During antral follicle development mouse oocytes undergo rearrangement of granulosa cell interactions and the oocytes released from follicles at the beginning or at the end of antral development are either devoid of denuded oocytes (DO) or strictly associated with cumulus-intact (CI) cumulus cells. In this study, these two oocyte classes were analyzed before germinal vesicle (GV) and after in vitro maturation (IVM) to evaluate (a) the ultrastructural aspect of oolemma microvilli by scanning electron microscopy analysis and (b) specific morphological markers of differentiation (chromatin organization, mitochondria, cortical granules, microfilaments, and spindle of metaphase II- MII-). At GV stage, CI oocytes exhibited remarkable differences (a) in the oolemma microvillar ultrastructure and distribution with respect to DO and (b) in the chromatin organization that was typical of meiotically competent germ cells. By contrast, homogeneous patterns of distribution of mitochondria, cortical granules, and microfilaments characterized both the oocyte classes. At the end of culture, CI oocytes, even when matured without cumulus cells, reached more efficiently the MII stage and acquired an ultrastructural microvillous configuration different from DO. In addition, MII-arrested DO had a higher percentage of meiotic spindles with abnormal morphology in comparison with preovulatory oocytes, while cortical granule and microfilament patterns revealed no appreciable differences between the groups. With regard to mitochondria, a polarized distribution of these organelles was found in 82% of DO and in 97% of CI oocytes. These observations suggested that the achievement of the full antral follicle development is a condition for the acquisition of specific qualitative properties that are essential for the production of fertilizable oocytes, both in in vivo and in vitro models as well. PMID- 16718657 TI - Morphodynamics of ovarian follicles during oogenesis in mice. AB - In the mouse, oogonia enter the prophase of the first meiotic division and differentiate into oocyte while developing in the fetal ovary. Shortly after birth, all oocytes are arrested in the dictyate stage of late prophase in the developing follicles; a small number of follicles reach the ovulatory stage; the rest are lost by apoptosis. The resumption of meiotic division and nuclear progression to metaphase II (oocyte maturation) occur in the ovulatory follicles. In this article we review recent morphological data that have clarified how cytokines and glycosaminoglycans (GAGs) are involved in mouse follicular development, atresia, and maturation during oogenesis, as exogenous/endogenous factors. (1) Microvascular networks and angiogenic factors (epidermal growth factor; GAGs) are deeply involved in selective mouse oocyte growth beyond approximately 20-30 microm in diameter. (2) Gonadotropin-inducible neuronal apoptosis inhibitory protein may indirectly affect oocyte survival as a result of the inhibition of apoptotic granulosa-cell death during folliculogenesis. (3) The pattern of oocyte degeneration depends on follicle and oocyte developmental stages, and follicle stimulating hormone accelerates the process of degeneration of oocytes. (4) The process of degeneration of mouse oocytes/eggs is modulated by tumor necrosis factor-alpha that is accumulated in the expanded cumulus during oocyte maturation. (5) A colloidal iron-positive substance was detected in the intercellular spaces of follicular tissue, especially in the cumulus mass. Cells located where the cumulus mass and granulosa cell layer interwound became enlarged during the resumption of oocyte meiosis. Colloidal iron-positive substances accumulated extensively within the intercellular spaces of the enlarged cells. PMID- 16718658 TI - Oocyte-follicle cell interactions during ovarian follicle development, as seen by high resolution scanning and transmission electron microscopy in humans. AB - The aim of this article is to summarize and update, through an integrated analysis by transmission electron microscopy (TEM) and high resolution scanning electron microscopy (SEM) after osmium-dimethyl sulfoxide-osmium (ODO) maceration, the studies of our research group on the morphodynamics of oocyte follicle cell associations during follicle development in humans. In resting oocytes, follicular cells project few and short cytoplasmic processes in the perioocytic space. They often form bulbous terminals very close to the oolemma where zonulae adherentes, maculae adherentes, and gap junctions are present. The oolemma mostly appears smooth with short and scanty microvilli. In early growing follicles, follicular cell projections appear as (a) long and tortuous microvilli or (b) large and short extensions. The oolemma shows numerous short microvilli. By TEM, long and thin follicular "intraooplasmic processes" have been seen to penetrate deeply into some oolemma invaginations. In macerated samples, they are observed by SEM to come very close to the nucleus and contact different oocyte organelles. These processes are more likely involved in early oocyte growth. In late growing follicles, oocyte-somatic cell interactions-now established through the interposition of the zona pellucida (ZP)-preserve the general features of early growth stage, with the exceptions of "intraooplasmic processes," which are no more present. In mature follicles subjected to a long ODO maceration, corona cells appear to contact the oocyte through an apical plume of numerous very long "curly hair-like microvilli." Corona cell microvilli, quite likely provide a sort of cytoplasmic skeleton for the ZP and they are possibly involved in (a) release of nutrients or removal catabolites to/from oocyte and vice versa and (b) transfer of substances to build up ZP. In conclusion, among oocyte and somatic cells a structural and functional association is revealed. This association, certainly highly dynamic in vivo, plays a key role in regulating the healthy folliculogenesis to assure a correct and timed oocyte maturation and ovulation. PMID- 16718659 TI - Morphological patterns of angiogenesis in ovarian follicle capillary networks. A scanning electron microscopy study of corrosion cast. AB - To describe the morphological characteristics of the ovarian follicle (F) capillary neoformation and regression, the angiogenic figures were studied by means of scanning electron microscopy of corrosion casts in developing and mature F of rabbit, pig, and cow. Developing F showed gradual neoformation of thecal capillaries characterized by budding and then sprouting, likely from preexisting interstitial vessels. Postcapillary venules frequently showed vasoconstriction rings (sphincters). Vasodilation followed capillary elongation. Mature F, in addition to vessel elongation and dilation, also presented infolding of dilated capillary walls, followed by capillary duplication and sinusoidalization. Periovulatory F mainly showed functional changes, such as capillary dilation, signs of iperpermeabilization, and ischemia, the latter being limited to the apical follicular area. Vessel regression was characterized by thinning of capillaries and presence of avascular areas within the atretic F wall at any stage. This study showed two main types of angiogenic patterns. (a) longitudinal elongation (in series, sprouting angiogenesis) characterizing the initial phase of F development and (b) parallel duplication (in parallel, infolding or intussusceptive angiogenesis), ending in capillary lateral replication or splitting, secondary to functional microvascular changes. Indirect evidence of the establishment of postcapillary resistances contributing to capillary remodeling, was also shown. It is concluded that the sequence of capillary neoformation in mammalian ovarian F occurs in six steps: (1) budding, (2) sprouting (and elongation), (3) dilation, (4) infolding (intussusception), (5) duplication (splitting and elongation), and (6) sinusoidalization. Capillary regression hits F at any stage and characterizes F atresia. PMID- 16718660 TI - Structural changes and cell properties of human ovarian surface epithelium in ovarian pathophysiology. AB - The surface epithelial cells of the ovary, which are modified peritoneal cells, form a single, focally pseudostratified layer. The Mullerian ducts differentiate after invagination of the coelomic mesothelium over the gonadal ridges during the 6th week of embryonic life. On the basis of the embryologically putative Mullerian potential of this epithelium, endometriosis can be explained by coelomic metaplasia from the peritoneum, including ovarian surface epithelium. Some pelvic endometriosis specimens have shown that epithelial cells on the ovary or pelvis are serially changed to endometriotic gland cells. Immunohistochemistry as well as scanning electron microscopy also reinforce the light-microscopical findings. A three-dimensional culture system demonstrated that human ovarian surface epithelial cells exhibited a glandular-stromal structure when they were cocultured with endometrial stromal cells in an estrogen-rich environment. Ovarian carcinomas in the epithelial-stromal category are thought to arise from the surface epithelium and its inclusions. The ovarian surface epithelium is physiologically involved in follicular rupture, oocyte release, and the subsequent repair of follicle wall during reproductive age. Simultaneously, ovulation may cause a loss of integrity of the surface epithelium, followed by accumulation of multiple mutations. The cortical invagination, surface stromal proliferation, and Mullerian differentiation of these cells are likely not to be an early step in the cancer development. However, the inclusion cysts are closely related with carcinogenesis because they are significantly more common in ovaries contralateral to those containing epithelial cancers than in control ovaries. As an in vitro study, ovarian carcinoma cell lines were established from simian virus 40 large T antigen-transformed human surface epithelial cells of the ovary. Further investigations of these cell lines may lead to insights into the preneoplastic and early stages of carcinomas. To clarify the pathogenesis of endometriosis and epithelial ovarian cancer, specifically designed studies of ovarian surface epithelium are required. PMID- 16718662 TI - Migration of human and mouse primordial germ cells and colonization of the developing ovary: an ultrastructural and cytochemical study. AB - This review is an account of the origin and migratory events of primordial germ cells until their settlement in the gonad before sexual differentiation in the human as well as mice. In this context, the morphodynamic characteristics of the migration of the primordial germ cells, the macromolecular characteristics of the extracellular matrix of the migratory pathway, and the factors involved in the germ cell guidance have been analyzed and discussed in the light of recent advances in this field, by means of immunocytochemical procedures. The events prior to gonadal morphogenesis and the origin of the somatic cell content of the human gonadal primordium have been also analyzed. In particular, evidences are presented showing that cells derived from the coelomic epithelium and mesenchyme are at the origin of the somatic components of the gonadal primordium, and that a mesonephric cell contribution to the generation of somatic cell components of the genital ridge in humans should be discarded due to the morphological stability of the different nephric structures during the period preceding the sexual differentiation of the gonad. PMID- 16718663 TI - Comparison of bone formation ingrafted periosteum harvested from tibia and calvaria. AB - Periosteum covers the bone surface and displays the potential to initiate bone formation, after injury to the bone. Numerous studies have demonstrated that the periosteum plays major roles in the healing process after bone fracture. Some reports have described that in the healing of long bone fractures, the periosteum forms new bone by intramembranous and endochondral ossification. Other researchers insist that healing of defects in membrane bone shows bone formation by intramembranous ossification. However, previous studies have not been able to clarify differences in bone formation patterns. We hypothesized that differences in bone formation pattern are associated with the periosteal potential for cell differentiation. The present study grafted periosteum, harvested from the tibia and calvaria, into the suprahyoid muscle, with the aim of interrupting release of factors from bone matrix. Bone formation, after grafting periosteum, harvested from the tibia and calvaria, was examined histologically and radiographically. Grafted tibial periosteum formed a large area of new bone by intramembranous and endochondral ossification, while grafted calvarial periosteum displayed intramembranous ossification. Grafted tibial periosteum formed a larger area of bone than grafted calvarial periosteum. Patterns of cell differentiation thus differ between grafted periosteum, harvested from the tibia and calvaria. PMID- 16718664 TI - Histological changes in rat nasal epithelia after unilateral neonatal naris occlusion. AB - The authors studied the extent of the different epithelia lining the nasal fossae of the albino rat after neonatal closure of one naris. Newborn pups were anesthetized by hypothermia and the external opening of their right naris cauterized, while littermates served as controls. Animals were sacrificed at 3 months, and the occluded (OCF) and nonoccluded (NOF) fossae of experimental animals as well as both fossae of control animals (CTF) were histologically studied. In both control and experimental animals, nasal fossae were lined by five different types of epithelia: squamous stratified, transitional, metaplastic, respiratory, and olfactory epithelia. It was found that closure of one naris provokes reorganization of the epithelial lining in both the occluded and nonoccluded side. In CTF airflow, physical conditions as well as pollutants and biological agents irritate the epithelial lining, causing squamous metaplasia as well as metaplastic epithelium showing inflammation in rostral levels. In CTF caudal levels, the metaplastic epithelium appears to a lesser degree and the respiratory epithelium prevails, except for the most caudal level where the olfactory epithelium is prevalent. In OCF, the protected environment created prevents the occurrence of metaplastic epithelium, the transitional, respiratory, and olfactory epithelia developing in the corresponding area instead. In NOF, where the airflow is double, the same pattern occurs as in CTF, although metaplastic epithelium values are approximately double, suggesting a clear linear effect. An outstanding feature observed was the increased extent of the olfactory epithelium in OCF regarding NOF, although changes in its morphological structure were not found. Airflow properties, including pressure, coldness, velocity, and turbulence, as well as biological and chemical hazards present in inflow, cause histological reorganization of the nasal epithelium lining during postnatal development. Results prove the need to consider airflow changes in nasal fossae surgery and point to the protective value of naris closure in ENT clinics, supporting it as a treatment of atrophic rhinitis. PMID- 16718665 TI - Electronic structure analyses of BN network materials using high energy resolution spectroscopy methods based on transmission electron microscopy. AB - Electronic structures of boron-nitride (BN) nanotubes and a BN cone-structure material were studied by using a high energy-resolution electron energy-loss spectroscopy (EELS) microscope. A trial of the whole electronic structure study of hexagonal BN (h-BN), which consists of flat BN honeycomb layers, was conducted by a combination of EELS and X-ray emission spectroscopy (XES) based on transmission electron microscopy (TEM) (TEM-EELS/XES). The pi and pi+sigma plasmon energies of BN nanotubes (BNT) were smaller than those of h-BN. The pi+sigma energy was explained by the surface plasmon excitation. The spectrum of a two-wall BNT of 2.7 nm in diameter showed a new spectral onset at 4 eV. The valence electron excitation spectra obtained from the tip region of the BN cone with an apex angle of 20 degrees showed similar intensity distribution with those of BNTs. The B K-shell electron excitation spectra obtained from the bottom edge region of the BN cone showed additional peak intensity when compared with those of h-BN and BNT. The B K-shell electron excitation spectra and B K-emission spectra of h-BN were compared with a result of a LDA band calculation. It showed that high symmetry points in the band diagram appear as peak and/or shoulder structures in the EELS and XES spectra. Interband transitions appeared in the imaginary part of the dielectric function of h-BN experimentally obtained were assigned in the band diagram. The analysis also presented that the LDA calculation estimated the bandgap energy smaller than the real material by an amount of 2 eV. Those results of TEM-EELS/XES analysis presented that high energy resolution spectroscopy methods combined with TEM is a promising method to analyze whole electronic structures of nanometer scale materials. PMID- 16718667 TI - Role of granulosa and theca cell interactions in ovarian follicular maturation. AB - We developed a culture system in which two types of ovarian follicular cells were allowed to attach to opposite sides of a collagen membrane. Using this in vitro cell culture system, we studied the effects of granulosa- and theca-cell interaction on the morphology, structure, and function of bovine ovarian follicular cells. In the first part of the study, we explored how the interaction between theca and granulosa cells affects the morphology and structure of the cells. This study was done using follicular cells collected from bovine ovarian follicles at the early developmental stage. Granulosa cells cultured alone were flattened, and formed a monolayer sheet. By contrast, granulosa cells cultured with theca cells were convex, and formed multilayer sheets. Theca cells cultured alone were thin, flat, and spindle-shaped. Theca cells cultured with granulosa cells were also spindle-shaped; however, they appeared convex and more densely packed when compared with theca cells cultured alone. In the second part of the study, the possible role of the cellular interaction in the control of differentiation and growth of granulosa and theca cells was investigated. When follicular cells were isolated from the early stage of follicular development, theca cells reduced progesterone and inhibin production by granulosa cells and augmented the growth of granulosa cells. When the cells were isolated from the late stage of follicular development, by contrast, theca cells augmented hormonal production by granulosa cells, and did not affect the growth of granulosa cells. The growth and androstenedione production by theca cells were increased by the presence of granulosa cells, irrespective of the origin of follicular cells. These results demonstrated that communication between two types of follicular cells results in reciprocal modulation of their morphology, structure, growth, and function. Cellular interactions seem to be one of the major factors controlling the differentiation and growth of the follicular cells during the follicular maturation process. In contrast to granulosa and theca cells cultured alone, cells in the coculture seemed to possess morphological and functional characteristics more similar to those of cells in the growing follicular wall in vivo. Thus, we speculate that the interaction between these two types of follicular cells is essential for the maintenance of original structure and function of the bovine follicular wall. PMID- 16718668 TI - Ultrastructural characteristics of human granulosa cells in a coculture system for in vitro fertilization. AB - The use of somatic cells for cocultures during in vitro fertilization (IVF) is currently finalized to obtain a higher number of healthy and viable embryos with a high potential of implantation. Among the different cell lines that can be used as feeder cells for cocultures, granulosa cells (GCs) are autologous cells, safe and easy to recover. The aim of the present study was to analyze the fine structure of human GCs used in a coculture system to evaluate, from a morphodynamic point of view, their role in supporting embryo development. GCs were collected during oocyte pick-up, 36 h after human chorionic gonadotropin administration, from patients undergoing IVF procedures, who had given their informed consent to be included in this protocol. After coculture, GCs were fixed and processed for light microscopy (LM) and transmission electron microscopy (TEM). By LM, GCs appeared as clusters of loosely packed cells, irregularly rounded or polyhedral in shape, varying in diameter from 18 to 25 microm. Mitotic cells, as well as regressing elements (with pyknotic nuclei or dense cytoplasm) and cell fragments were occasionally observed. By TEM, the plasma membrane was irregular due to the presence of cytoplasmic evaginations. Linear and annular gap junctions between neighboring GCs were found. GC nuclei, rounded and eccentrically located, contained finely dispersed chromatin, one (often two) prominent nucleoli and, infrequently, peripheral patches of heterochromatin. Numerous organelles populated the GC cytoplasm, among them, mitochondria were rod shaped or elongated, usually provided with tubular-vesicular cristae but occasionally showing atypical, longitudinally oriented cristae. Membranes of smooth endoplasmic reticulum, Golgi stacks and vesicles, secretory-like granules, cisternae of rough endoplasmic reticulum (RER), free ribosomes and polysomes, lysosomal-like bodies, microfilaments, and lipid droplets were also seen in the GC cytoplasm. In most cells, RER was scarcely represented and numerous lipid droplets filled the perinuclear space. On the contrary, some GCs contained an abundant RER and rare lipid droplets scattered in the cytoplasm. In conclusion, our data demonstrated the presence, in a coculture system, of GCs provided with ultrastructural characteristics typical of healthy, metabolically active, mostly steroidogenic cells. Protein-synthetic cells have also been detected. These data, evaluated at the light of biochemical and clinical studies, sustain the capability of human GCs cocultures to positively affect early embryo development in vitro by the secretion of steroids and proteins, putative "embryotrophic" factors. PMID- 16718669 TI - Unified structural equation modeling approach for the analysis of multisubject, multivariate functional MRI data. AB - The ultimate goal of brain connectivity studies is to propose, test, modify, and compare certain directional brain pathways. Path analysis or structural equation modeling (SEM) is an ideal statistical method for such studies. In this work, we propose a two-stage unified SEM plus GLM (General Linear Model) approach for the analysis of multisubject, multivariate functional magnetic resonance imaging (fMRI) time series data with subject-level covariates. In Stage 1, we analyze the fMRI multivariate time series for each subject individually via a unified SEM model by combining longitudinal pathways represented by a multivariate autoregressive (MAR) model, and contemporaneous pathways represented by a conventional SEM. In Stage 2, the resulting subject-level path coefficients are merged with subject-level covariates such as gender, age, IQ, etc., to examine the impact of these covariates on effective connectivity via a GLM. Our approach is exemplified via the analysis of an fMRI visual attention experiment. Furthermore, the significant path network from the unified SEM analysis is compared to that from a conventional SEM analysis without incorporating the longitudinal information as well as that from a Dynamic Causal Modeling (DCM) approach. PMID- 16718671 TI - Adult weight gain and histopathologic characteristics of breast cancer among postmenopausal women. AB - BACKGROUND: Although the link between postmenopausal breast cancer and adiposity is well established, the association between weight gain and specific histopathologic characteristics of breast carcinoma has not been studied carefully. METHODS: Using 1200 incident invasive breast cancers among 44,161 postmenopausal women who were not taking hormone therapy in the American Cancer Society's Cancer Prevention Study II Nutrition Cohort, the authors computed age adjusted rates and rate ratios (RR) for breast cancer by histology, stage, grade, and estrogen receptor (ER) and progesterone receptor (PR) status by categories of adult weight gain. RESULTS: Age-adjusted rates of breast cancer were highest for women who reported the most weight gain, regardless of histologic type. For weight gain >60 pounds, compared with weight gain < or =20 pounds the RR for ductal carcinoma was 1.89 (95% confidence interval [95%CI], 1.53-2.34), and the RR for lobular carcinoma was 1.54 (95%CI. 1.01-2.33). Weight gain was associated with increased risk at every tumor stage and grade. The risk for regional or distant stage was elevated significantly in every category of weight gain and was 3 times higher among women who had the greatest weight gain (RR, 3.15; 95%CI, 2.21-4.48). Weight gain was associated with increased risk of ER-positive/PR positive tumors (P for trend <.0001) but not ER-negative/PR-negative tumors (P for trend = .09). The results essentially remained unchanged when the analysis was restricted to women who had regular screening mammograms. CONCLUSIONS: Excess adiposity is an important contributor to breast cancer risk among postmenopausal women, regardless of histologic type, and especially for tumors of advanced stage and high grade. PMID- 16718672 TI - The fragmentation pathways of protonated Amiton in the gas phase: towards the structural characterisation of organophosphorus chemical warfare agents by electrospray ionisation tandem mass spectrometry. AB - Amiton (O,O-diethyl-S-[2-(diethylamino)ethyl] phosphorothiolate), otherwise known as VG, is listed in schedule 2 of the Chemical Weapons Convention (CWC) and has a structure closely related to VX (O-ethyl-S-(2 diisopropylamino)ethylmethylphosphonothiolate). Fragmentation of protonated VG in the gas phase was performed using electrospray ionisation ion trap mass spectrometry (ESI-ITMS) and revealed several characteristic product ions. Quantum chemical calculations provide the most probable structures for these ions as well as the likely unimolecular mechanisms by which they are formed. The decomposition pathways predicted by computation are consistent with deuterium-labeling studies. The combination of experimental and theoretical data suggests that the fragmentation pathways of VG and analogous organophosphorus nerve agents, such as VX and Russian VX, are predictable and thus ESI tandem mass spectrometry is a powerful tool for the verification of unknown compounds listed in the CWC. PMID- 16718670 TI - Inhibitory effects of triterpenes and flavonoids on the enzymatic activity of hyaluronic acid-splitting enzymes. AB - The effect of triterpenes and flavonoids on the activity of several hyaluronic acid-splitting enzymes was investigated. Studies showed that the inhibitory effect of the triterpenes glycyrrhizin and glycyrrhetinic acid is dependent on the source of hyaluronate lyase. Hyaluronate lyase from Streptococcus agalactiae (Hyal B) and recombinant hyaluronate lyase from Streptococcus agalactiae (rHyal B) demonstrated strongest inhibition. In contrast, hyaluronate lyases from Streptomyces hyalurolyticus (Hyal S), Streptococcus equisimilis (Hyal C) and hyaluronidase from bovine testis (Dase) showed only reduced inhibition action. A non-competitive dead end inhibition with Ki=3.1+/-1.8x10(-6) mol/mL and Kii=6.7+/ 2.4x10(-6) mol/mL was found for glycyrrhizin on recombinant hyaluronate lyase from Streptococcus agalactiae. The inhibitory effect of flavonoids on Hyal B, rHyal B and Dase was determined depending on the number of hydroxyl groups and side chain substituents in the molecule. Flavonoids with many hydroxyl groups inhibited hyaluronate lyase stronger than those with only a few. Native hyaluronate lyase (Hyal B) showed a more extensive inhibition than the recombinant protein (rHyal B). Accordingly, the inhibition by triterpenes and flavonoids is presumably specific for each hyaluronic acid (HA)-splitting enzyme. PMID- 16718673 TI - Characterization of high explosive particles using cluster secondary ion mass spectrometry. AB - The use of secondary ion mass spectrometry (SIMS) for the detection and spatially resolved analysis of individual high explosive particles is described. A C(8) (-) carbon cluster primary ion beam was used in a commercial SIMS instrument to analyze samples of high explosives dispersed as particles on silicon substrates. In comparison with monatomic primary ion bombardment, the carbon cluster primary ion beam was found to greatly enhance characteristic secondary ion signals from the explosive compounds while causing minimal beam-induced degradation. The resistance of these compounds to degradation under ion bombardment allows explosive particles to be analyzed under high primary ion dose bombardment (dynamic SIMS) conditions, facilitating the rapid acquisition of spatially resolved molecular information. The use of cluster SIMS combined with computer control of the sample stage position allows for the automated identification and counting of explosive particle distributions on silicon surfaces. This will be useful for characterizing the efficiency of transfer of particulates in trace explosive detection portal collectors and/or swipes utilized for ion mobility spectrometry applications. PMID- 16718675 TI - Problems in the naming of genes. PMID- 16718674 TI - Interception of the radicals produced in electrophilic fluorination with radical traps (Tempo, Dmpo) studied by electrospray ionization mass spectrometry. AB - The interaction of the nitroxide radical traps (Tempo and Dmpo) and radicals produced in the electrophilic fluorination of olefins (styrene and alpha methylstyrene) and Selectfluor (1-chloromethyl-4-fluoro-1,4 diazoniabicyclo[2.2.2]octanebis(tetrafluoroborate) (F-TEDA-BF(4)) (1)) was investigated by electrospray ionization mass spectrometry (ESI-MS). Tempo succeeded in intercepting the radical cationic intermediates and the radical adduct ions were detected at m/z 260 (for styrene) and m/z 274 (for alpha methylstyrene). Dmpo could also intercept the fluorine radical and radical adduct ions were detected at m/z 131, 132 and 152. The interception of the radical cationic intermediates and fluorine radical is good evidence for the presence of a single-electron transfer mechanism in the electrophilic fluorination. PMID- 16718676 TI - Holoprosencephaly: clinical evaluation on audiological and brainstem electrophysiological profiles. AB - This study evaluated audiological and electrophysiological profiles in 13 patients with holoprosencephaly. All patients had imaging evaluation by magnetic resonance imaging and molecular screening for the genes SHH, GLI2, and SIX3. Each patient underwent clinical (otological and vestibular antecedents, otoscopy) and instrumental (tympanometry, auditory brainstem response--ABR) evaluation to compare hearing and the electrophysiological profile possibly occurring in patients with these mutations. To our knowledge there are no systematic studies correlating molecular/imaging and evoked potentials in patients with HPE. Here, we discuss the audiological and electrophysiological profiles of patients and the possible role of the genes studied on the overall findings. PMID- 16718677 TI - Polyol synthesis in Aspergillus niger: influence of oxygen availability, carbon and nitrogen sources on the metabolism. AB - Polyol production has been studied in Aspergillus niger under different conditions. Fermentations have been run using high concentration of glucose or xylose as carbon source and ammonium or nitrate as nitrogen source. The growth of biomass, as freely dispersed hyphae, led to an increase of medium viscosity and hereby a decrease in mass transfer, especially oxygen transfer. The consequence was a decrease in DOT and the occurrence of a switch between fully aerobic conditions and oxygen-limited conditions. Metabolite quantification showed that polyols were the main metabolic products formed and represented up to 22% of the carbon consumed in oxygen-limited conditions. The polyol concentration and the polyol pattern depended strongly on the environmental conditions. This is due to a complex regulation of polyol production and to the fact that each polyol can fulfill different functions. In this study, erythritol, xylitol, and arabitol were produced as carbon storage compounds when the flux through the PP pathway exceeded the need in ribulose-5-phosphate for the biomass synthesis. Glycerol, erythritol, and xylitol seem to be involved in osmoregulation. Mannitol was produced when the catabolic reduction of charge was high. Its production involves the enzyme NAD-dependent mannitol-1-phosphate dehydrogenase and seems to be the main cytosolic route for the NADH reoxidation during oxygen limitation. PMID- 16718678 TI - Engineering Escherichia coli to improve culture performance and reduce formation of by-products during recombinant protein production under transient intermittent anaerobic conditions. AB - Three E. coli strains, named VAL22, VAL23, and VAL24, were engineered at the level of mixed-acid fermentation pathways to improve culture performance under transient anaerobic conditions. VAL22 is a single mutant with an inactivated poxB gene that codes for pyruvate oxidase which converts pyruvate to acetate. VAL23 is a double mutant unable to produce lactate and formate due to deletions of the ldhA and pflB genes that code for lactate dehydrogenase and pyruvate-formate lyase, respectively. VAL24 is a triple mutant with ldhA and pflB deleted and poxB inactivated. Engineered strains were cultured under oscillating dissolved oxygen tension (DOT) in a scale-down system, to simulate gradients occurring in large scale bioreactors. Kinetic and stoichiometric parameters of constant (10%) and oscillating DOT cultures of the engineered strains were compared with those of the parental strain, W3110. All strains expressed recombinant green fluorescent protein (GFP) as a protein model. Mutant strains showed improved specific growth rate, reduced by-product formation, and reduced specific glucose uptake rate compared to the parental strain, when cultured under oscillating DOT. In particular, lactate and formate production was abolished and acetate accumulation was reduced by 9-12%s. VAL24 showed the best performance, as specific growth and GFP production rates, and maximum GFP concentration were not affected by DOT gradients and were at least twofold higher than those of W3110 under constant DOT. Under oscillating DOT, VAL24 wasted about 40% less carbon into fermentation by-products than W3110. It was demonstrated that, although E. coli responds rapidly to DOT fluctuations by deviating to fermentative metabolism, such pathways can be eliminated as they are not necessary for bacterial survival during the short circulation times typical of large-scale cultures. The approach shown here opens new possibilities for designing metabolically engineered strains, with reduced sensitivity to DOT gradients and improved performance under typical conditions of large-scale cultures. PMID- 16718679 TI - Nanogels prepared by self-assembly of oppositely charged globular proteins. AB - Ovalbumin and lysozyme are two main proteins in hen egg white with the isoelectric points of 4.8 and 11, respectively. Herein we report the manufacture of stable, narrowly distributed nanogels (hydrodynamic radius about 100 nm) using a novel and convenient method: ovalbumin and lysozyme solutions were mixed at pH 5.3, the mixture solution was adjusted to pH 10.3, then subsequently stirred and heated. The nanogels were characterized using a combination of techniques. The nanogels have spherical shape and core-shell structure. The core is mainly composed of lysozyme and the shell is mainly composed of ovalbumin. The proteins in the nanogels are in denatured states and they are bound by intermolecular hydrophobic interactions, hydrogen bonds, and disulfide bonds. The charges of the nanogels can be modulated by the pH of the medium. The electrostatic repulsion of ovalbumin molecules on the nanogel surface stabilizes the nanogels in aqueous solution. The formation mechanism of the nanogels is discussed. PMID- 16718681 TI - Continuous enzymatic transesterification of sesame oil and a fully hydrogenated fat: effects of reaction conditions on product characteristics. AB - An immobilized lipase from Thermomyces lanuginosus (TL IM) was employed to mediate the continuous transesterification of sesame oil and fully hydrogenated soybean oil (FHSBO) in a packed-bed reactor operating at 70 degrees C. Reactions between sesame oil (rich in LLL (15.97%), LOL (31.56%), and OLO (21.15%) [L = linoleic; O = oleic]) and the fully hydrogenated fat ((73.7% SSS, 26.3% SPS) [S = stearic; P = palmitic]) produced semi-solid fats. These products are complex mixtures of triacylglycerol (TAG) species whose compositions depend on reaction conditions. The dependence of the steady state product TAG profile on space time was determined for four initial weight ratios of sesame oil to hydrogenated fat (90:10, 80:20, 70:30, and 60:40). Except for the trial involving a weight ratio of sesame oil to FHSBO of 60:40, near equilibrium conditions were achieved at space times of 30 min-1 h. The chemical, physical, and functional properties of the product semi-solid fats were characterized. The predominant TAG species in the quasi-equilibrium products obtained from the mixture initially containing 90% (w/w) sesame oil and 10% FHSBO were LOL (26.22%) and OLO (21.92%). For transesterification of 80% sesame oil and 20% FHSBO, the major product species were OOP (21.27%), LOL (17.46%), and OLO (13.93%). OOP (24.38%) was the major product for reaction of 70% sesame oil with 30% FHSBO. Appropriate choices of reaction conditions and initial ratios of sesame oil to FHSBO lead to TAG with melting profiles and solid fat contents (SFC) similar to those of a variety of commercial products. PMID- 16718682 TI - Cross-linked N-telopeptide of type I collagen (NTx) in urine as a predictor of periprosthetic osteolysis. AB - Periprosthetic osteolysis is often nonsymptomatic and hard to visualize by conventional radiography. Cross-linked N-telopeptide of type I collagen (NTx), a marker of osteoclast mediated bone resorption, has been suggested to evaluate local particulate-induced osteolysis in patients operated on with a total hip prosthesis. Urine specimens were sampled after hip joint replacement in 160 patients. NTx was analyzed by a commercially available ELISA kit. Osteolysis was identified in the acetabulum and confirmed at operation. Using analysis of covariance to correct for differences in age, gender, and time after operation, NTx (mean SD) was 36+/-12 BCE/nM creatinine in patients with osteolysis (n=33) and 27+/-13 BCE/nM creatinine in patients without osteolysis (n=127) (p=0.003). Eighteen hips of 38 (47%), demonstrating an annual wear of more than 0.2 mm and an NTx value above 29 BCE/nM creatinine, had been revised due to osteolysis. The osteolysis prevalence in this group was increased 10 times (CI 4-23, p<0.05). Indeed, NTx release and annual wear were both associated with increased prevalence of osteolysis, however, independently of each other. NTx seems a feasible marker of periprosthetic osteolysis. A preoperative baseline NTx level is likely needed for its use as a predictor of periprosthetic osteolysis in individual cases. PMID- 16718683 TI - Axonal degeneration stimulates the formation of NG2+ cells and oligodendrocytes in the mouse. AB - Proliferation of the adult NG2-expressing oligodendrocyte precursor cells has traditionally been viewed as a remyelination response ensuing from destruction of myelin and oligodendrocytes, and not to the axonal pathology that is also a characteristic of demyelinating disease. To better understand the response of the NG2+ cells to the different components of demyelinating pathology, we investigated the response of adult NG2+ cells to axonal degeneration in the absence of primary myelin or oligodendrocyte pathology. Axonal degeneration was induced in the hippocampal dentate gyrus of adult mice by transection of the entorhino-dentate perforant path projection. The acutely induced degeneration of axons and terminals resulted in a prompt response of NG2+ cells, consisting of morphological transformation, cellular proliferation, and upregulation of NG2 expression days 2-3 after surgery. This was followed by a reduction of cellular NG2 expression to subnormal levels from day 5 to 7 and reappearance of normal appearing NG2+ cells from day 10. Mice that had received repeated injections of bromodeoxyuridine from 24 to 72 h after surgery contained significant numbers of bromodeoxyuridine-incorporating oligodendrocytes in the areas with axonal degeneration at day 7. The results suggest that axonal degeneration induces a unique sequence of changes of NG2+ cells and that a subpopulation of the newly generated NG2+ cells differentiate into oligodendrocytes. PMID- 16718684 TI - Hydrogen sulphide regulates calcium homeostasis in microglial cells. AB - Hydrogen sulphide (H2S), which is produced endogenously from L-cysteine in mammalian tissues, has been suggested to function as a neuromodulator in the brain. However, the role of H2S in microglial cells is unclear. In this study, the effect of exogenous and endogenous H2S on intracellular calcium homeostasis was investigated in primary cultured microglial cells. Sodium hydrosulphide (NaHS), a H2S donor, caused a concentration-dependent (0.1-0.5 mM) increase in intracellular calcium concentration ([Ca2+]i). This effect was significantly attenuated in the presence of a calcium-free extracellular solution, Gd3+ (100 microM), a nonselective Ca2+ channel blocker, or thapsigargin (2 microM), an inhibitor of the sarcoplasmic/endoplasmic reticulum Ca2+ -ATPase. These observations suggest that the increase in [Ca2+]i in response to H2S involves both calcium influx across the plasma membrane and calcium release from intracellular stores. The H2S-induced calcium elevation is partly attenuated by H 89, a selective cAMP-dependent protein kinase (PKA) inhibitor, but not by U73122, a phospholipase C (PLC) inhibitor, and chelerythrine, a selective protein kinase C (PKC) inhibitor, suggesting the involvement of cAMP/PKA, but not PLC/PKC/phosphoinositol-3,4,5-inositol (IP3) pathway. Using RT-PCR, only cystathionine gamma-lyase (CSE), a H2S producing enzyme, was detected in primary cultures of microglia. Lowering endogenous H2S level with, D,L-propargylglycine and beta-cyano-L-alanine, two CSE inhibitors, significantly decreased [Ca2+]i, suggesting that endogenous H2S may have a positive tonic influence on [Ca2+]i homeostasis. These findings support the possibility that H2S may serve as a neuromodulator to facilitate signaling between neurons and microglial cells. PMID- 16718685 TI - Tolbutamide reduces glioma cell proliferation by increasing connexin43, which promotes the up-regulation of p21 and p27 and subsequent changes in retinoblastoma phosphorylation. AB - Our previous work has shown that tolbutamide increases gap junctional permeability in poorly coupled C6 glioma cells and that this effect is similar and additive to that found with dbcAMP, a well-known activator of gap junctional communication. Furthermore, the increase in gap junctional communication promoted by tolbutamide or dbcAMP is concurrent with the inhibition of proliferation of C6 glioma cells. In the present work, we show that tolbutamide and dbcAMP increase the synthesis of the tumor suppressor protein Cx43 and that they decrease the level of Ki-67, a protein expressed when cells are proliferating. These effects were accompanied by a reduction in the phosphorylation of pRb, mainly on Ser-795, a residue critical for the control of cell proliferation. The decrease in the phosphorylation of pRb is not likely to be mediated by a reduction in the levels of D-type cyclins, since instead of decreasing the expression of cyclins, D1 and D3 increased slightly after treatment with tolbutamide or dbcAMP. However, the Cdk inhibitors p21 and p27 were up-regulated after treatment with tolbutamide and dbcAMP, suggesting that they would be involved in the decrease in pRb phosphorylation. When Cx43 was silenced by siRNA, neither tolbutamide nor dbcAMP were able to up-regulate p21 and consequently to reduce glioma cell proliferation, as judged by Ki-67 expression. In conclusion, tolbutamide and dbcAMP inhibit C6-glioma cell proliferation by increasing Cx43, which correlates with a reduction in pRb phosphorylation due to the up-regulation of the Cdk inhibitors p21 and p27. PMID- 16718686 TI - Autosomal dominant atretic cephalocele with phenotype variability: report of a Brazilian family with six affected in four generations. AB - Atretic cephalocele is a clinicopathological entity, which is different from the common form of cephalocele. Its etiopathogenesis has not been completely explained and there are only two previous reports of familial recurrence. We report a Brazilian family with autosomal dominant inheritance with variable expressivity. PMID- 16718687 TI - Progressive nuclear factor-kappaB activation resistant to inhibition by contraction and curcumin in mdx mice. AB - Skeletal muscle of patients with Duchenne-type muscular dystrophy and mdx mice exhibits elevated activity of the transcription factor NF-kappaB (nuclear factor kappaB), which may play a role in muscle catabolism. We measured skeletal muscle NF-kappaB activity in mdx mice at three ages (10 days, 4 weeks, and 8 weeks) to test the hypothesis that NF-kappaB activity is elevated in an age-dependent manner in these mice. In addition, we tested the hypothesis that NF-kappaB activity could be reduced in mdx skeletal muscle by dietary supplementation with curcumin (1% w/v) or by fatiguing muscle contractions. We found that NF-kappaB activity was elevated at 4 and 8 weeks of age but not at 10 days, and was resistant to inhibition by either fatiguing contractions or dietary curcumin. We conclude that NF-kappaB activity is elevated in dystrophic skeletal muscle in an age-related manner and is resistant to inhibition by physiological and pharmacological means. These findings are consistent with a role for NF-kappaB activation in dystrophic muscle wasting but suggest that predicted interventions such as exercise or inhibitors of the early steps in the NF-kappa activation pathway may not be effective and that targeted research is needed to identify novel therapeutic strategies. PMID- 16718688 TI - Alterations in the gastrocnemius muscle of undernourished suckling rats. AB - Protein-energy undernutrition (PEU) during both the gestational and suckling periods causes a reduction in muscle fiber cross-section and a delay in fiber type differentiation. Such changes are not found when PEU occurs after the suckling period. To elucidate the consequences of PEU on muscle during the suckling period, we undernourished (UND) newborn rats by reducing their food intake, including a 6-hour fast every day, and doubling the number of pups suckled by the nursing rat. At day 24 of life, the gastrocnemius muscle showed a discrete reduction in lipids in the fibers of 5 control rats, and in 15 UND rats (P < 0.001). The type 2a and 2b fibers of the UND rats showed a smaller cross sectional area than control rats (P < 0.05). The distribution frequency histogram for the muscle fibers was skewed to the left in the undernourished rats, and this was significant for type 2a and 2b fibers. PEU during the suckling period may thus induce a delay in muscle fiber maturation but does not produce significant structural alterations. PMID- 16718689 TI - Detection of small-fiber neuropathy by sudomotor testing. AB - The symptoms of burning sensation affecting the feet, thought to be due to a distal small-fiber neuropathy (DSFN) affecting somatic unmyelinated fibers, are usually accompanied by vasomotor or sudomotor changes suggestive of involvement of autonomic fibers. We therefore examined the relationship between pattern of anhidrosis and DSFN and its etiology, comparing patients with "pure" DSFN (with normal nerve conduction) to those with clinical DSFN (minor conduction abnormalities). We reviewed 125 cases with a clinical phenotype of DSFN. These patients had distal burning discomfort, variable sensory deficits, and intact motor function. All had undergone assessment with thermoregulatory sweat test (TST), autonomic reflex screen (ARS), and nerve conduction studies and electromyography (NCS/EMG). TST showed a distal pattern of anhidrosis in 74%. The quantitative sudomotor axon reflex test (QSART) was abnormal in 74%, with 80% of those having a length-dependent pattern of anhidrosis/hypohidrosis. In total, 93% of patients had a distal pattern of abnormality on QSART or TST. The Composite Autonomic Severity Score (CASS) was used to quantify the severity and distribution of autonomic deficits: 98% had CASS abnormality (sudomotor, 98%; adrenergic, 43%; cardiovagal, 35%). EMG was normal or showed unrelated abnormalities in 75%. The most common etiologies of DSFN were idiopathic (73%), presumed hereditary (18%), and diabetes (10%). Sudomotor examination is thus a highly sensitive detection tool in DSFN. Autonomic involvement is mainly distal, and additionally may involve adrenergic and the long cardiovagal fibers. PMID- 16718690 TI - B lineage cells in the inflammatory central nervous system environment: migration, maintenance, local antibody production, and therapeutic modulation. AB - B cells have long played an enigmatic role in the scenario of multiple sclerosis pathogenesis. This review summarizes recent progress in our understanding of B cell trafficking, survival, and differentiation in the central nervous system (CNS). We propose four possible routes of intrathecal immunoglobulin-producing cells. The inflammatory CNS provides a unique, B-cell-friendly environment, in which B lineage cells, notably long-lived plasma cells, can survive for many years, perhaps even for a lifetime. These new findings offer a plausible explanation for the notorious persistence and stability of cerebrospinal fluid oligoclonal bands. Furthermore, we highlight similarities and differences of intrathecal immunoglobulin production in multiple sclerosis patients and patients with other CNS inflammatory conditions. Finally, we outline the possibly double edged effects of B cells and immunoglobulin in the CNS and discuss various therapeutic strategies for targeting the B-cell response. PMID- 16718691 TI - Risk factors for excessive sleepiness in older adults. AB - OBJECTIVE: To determine risk factors for excessive daytime sleepiness in older adults. METHODS: This is a cross-sectional study assessing multiple risk factors for excessive daytime sleepiness in older subjects (mean age, 78 years; range 65 98 years) with (n=149) and without (n=144) complaints of frequent excessive daytime sleepiness. Assessment of risk factors included full in-laboratory sleep studies. RESULTS: Excessive sleepiness among the elderly is multifactorial. Multivariable modeling identified the following as simultaneously significant risk factors for excessive sleepiness: severe sleep-disordered breathing (apnea hypopnea index, >30 episodes/hr), self-report of poor sleep quality, increased percentage of time in rapid eye movement sleep, pain at night at least three times per week, wheezing or whistling from chest at night, and medications with sleepiness as a side effect. Male sex also was associated with increased risk, whereas alcohol use (more than seven beverages per week) reduced the risk for sleepiness. Multiple risk factors were more commonly present in those with complaints of sleepiness. The presence of periodic limb movements, which are common in older adults, was not associated with sleepiness. INTERPRETATION: There is a distinct differential diagnosis of excessive daytime sleepiness in older adults. Many of the risk factors that we identified are treatable. PMID- 16718692 TI - Aerobic conditioning: an effective therapy in McArdle's disease. AB - OBJECTIVE: Susceptibility to exertional cramps and rhabdomyolysis in myophosphorylase deficiency (McArdle's disease [MD]) may lead patients to shun exercise. However, physical inactivity may worsen exercise intolerance by further reducing the limited oxidative capacity caused by blocked glycogenolysis. We investigated whether aerobic conditioning can safely improve exercise capacity in MD. METHODS: Eight MD patients (4 men and 4 women; age range, 33-61 years) pedalled a cycle ergometer for 30 to 40 minutes a day, 4 days a week, for 14 weeks, at an intensity corresponding to 60 to 70% of maximal heart rate. We monitored serum creatine kinase levels; changes in peak cycle work, oxygen uptake, and cardiac output; presence and magnitude of a spontaneous and glucose induced second wind; and citrate synthase and beta-hydroxyacyl coenzyme A dehydrogenase enzyme activities in quadriceps muscle. RESULTS: The prescribed exercise program increased average work capacity (36%), oxygen uptake (14%), cardiac output (15%), and citrate synthase and beta-hydroxyacyl coenzyme A dehydrogenase enzyme levels (80 and 62%, respectively) without causing pain or cramping or increasing serum creatine kinase. A spontaneous and glucose-induced second wind was present and was of similar magnitude in each patient before and after training. INTERPRETATION: Moderate aerobic exercise is an effective means of improving exercise capacity in MD by increasing circulatory delivery and mitochondrial metabolism of bloodborne fuels. PMID- 16718693 TI - Effects of neonatal dexamethasone treatment on hippocampal synaptic function. AB - OBJECTIVE: Synthetic glucocorticoid dexamethasone (DEX) is frequently used as a therapeutic agent to lessen the morbidity of chronic lung disease in premature infants. Surprisingly, little is known about the long-term neurodevelopmental outcomes of this therapy. METHODS: Using a schedule of tapering doses of DEX similar to that used in premature infants, we examined the consequences of neonatal DEX treatment on hippocampal synaptic plasticity of infants and associative memory later in their lives. RESULTS: Neonatal DEX treatment changed the direction of synaptic plasticity, favoring low-frequency, stimulation induced, long-term depression and opposing the induction of long-term potentiation by high-frequency stimulation in adolescent (5-week-old) rats, but these alterations disappeared in young adult (8-week-old) rats. The effects of DEX on long-term depression and long-term potentiation were found to correlate with an increase in the autophosphorylation of Ca(2+)/calmodulin-dependent protein kinase II and a decrease in the protein phosphatase 1 activity. Neonatal DEX treatment also disrupted memory retention in 5-week-old (but not 8-week-old) rats subjected to passive avoidance learning tasks. INTERPRETATION: These results suggest that neonatal DEX treatment alters hippocampal synaptic plasticity and contextual fear memory formation in later life, but these impairments apparently are not permanent. PMID- 16718694 TI - A mutation in the GABA(A) receptor alpha(1)-subunit is associated with absence epilepsy. AB - OBJECTIVE: To detect mutations in GABRA1 in idiopathic generalized epilepsy. METHODS: GABRA1 was sequenced in 98 unrelated idiopathic generalized epilepsy patients. Patch clamping and confocal imaging was performed in transfected mammalian cells. RESULTS: We identified the first GABRA1 mutation in a patient with childhood absence epilepsy. Functional studies showed no detectable GABA evoked currents for the mutant, truncated receptor, which was not integrated into the surface membrane. INTERPRETATION: We conclude that this de novo mutation can contribute to the cause of "sporadic" childhood absence epilepsy by a loss of function and haploinsufficiency of the GABA(A) receptor alpha(1)-subunit, and that GABRA1 mutations rarely are associated with idiopathic generalized epilepsy. PMID- 16718697 TI - Sleepiness in the elderly. PMID- 16718698 TI - Nerve center: events, people, and issues in academic neurology (interview with Eric R. Kandel). PMID- 16718699 TI - Novel missense mutation in ALS2 gene results in infantile ascending hereditary spastic paralysis. AB - OBJECTIVE: Recessive mutations in ALS2 (juvenile amyotrophic lateral sclerosis) are causative for early-onset upper motor neuron diseases, including infantile ascending hereditary spastic paralysis (IAHSP). The goal of this study is to identify novel disease-causing ALS2 mutations. METHODS: Mutations in ALS2 were screened by direct sequencing of complementary DNA obtained from patients' lymphoblasts. RESULTS: We report a novel ALS2 missense mutation in patients affected by IAHSP. This homozygous G669A mutation in exon 4 is predicted to result in a tyrosine substitution at cysteine 156 of the RCC1 (regulator of chromatin condensation)-like domain, encoding a putative guanine exchange factor for Ran guanosine triphosphatase, leading to a loss of ALS2 function due to instability of mutant protein. INTERPRETATION: These results highlight the important role of the RCC1-like domain in ALS2 stability and function that is essential for upper motor neuron maintenance. PMID- 16718700 TI - Reply: GDNF poses troubling questions for doctors, drug maker. PMID- 16718701 TI - Gene therapy of the brain in the dog model of Hurler's syndrome. AB - OBJECTIVE: A defect of the lysosomal enzyme alpha-L-iduronidase (IDUA) interrupts the degradation of glycosaminoglycans in mucopolysaccharidosis type I, causing severe neurological manifestations in children with Hurler's syndrome. Delivery of the missing enzyme through stereotactic injection of adeno-associated virus vectors coding for IDUA prevents neuropathology in affected mice. We examined the efficacy and the safety of this approach in enzyme-deficient dogs. METHODS: Because deficient dogs raise antibodies against IDUA in response to infusion, intracerebral vector injections were combined with an immunosuppressive regimen. RESULTS: Treatment was tolerated well. We observed broad dispersion of vector genomes in the brain of efficiently immunosuppressed dogs. The delivery of IDUA to large areas, which could encompass the entire brain, prevented glycosaminoglycan and secondary ganglioside accumulations. This condition was associated with drastic reduction of neuropathology throughout the encephalon. In contrast, vector injection combined with partial immunosuppression was associated with subacute encephalitis, production of antibodies against IDUA in brain tissues, and elimination of genetically modified cells. INTERPRETATION: Gene therapy directed to the entire brain is feasible and may be beneficial to children with Hurler's syndrome. The possibility of subacute encephalitis emphasizes the importance of preventing immune response against IDUA, a problem that needs to be considered in similar therapies for other genetic defects. PMID- 16718702 TI - Head injury and Parkinson's disease risk in twins. AB - OBJECTIVE: Head injury is an inconsistently reported risk factor for Parkinson's disease (PD). Many related variables might confound this association, such as differences in childhood and adolescent lifestyles or genetically determined risk taking behaviors. Twin studies circumvent some of these problems, because twins are genetically and environmentally much more similar than typical cases and control subjects. METHODS: We conducted a case-control study in 93 twin pairs discordant for PD ascertained from the National Academy of Sciences/National Research Council World War II Veteran Twins Cohort. RESULTS: A prior head injury with amnesia or loss of consciousness was associated with an increased risk for PD (odds ratio, 3.8; 95% confidence interval, 1.3-11; p = 0.014). Truncating observations 10 years before PD onset enhanced the association. Though less precise, the association was somewhat stronger in monozygotic than in dizygotic pairs. Risk increased further with a subsequent head injury (p trend = 0.022) and with head injuries requiring hospitalization. Duration of unconsciousness was not associated. In a subanalysis of 18 pairs concordant for PD, the twin with younger onset PD was more likely to have sustained a head injury, although numbers were small. INTERPRETATION: Our results suggest that mild-to-moderate closed head injury may increase PD risk decades later. PMID- 16718705 TI - Quantifying axonal loss after optic neuritis with optical coherence tomography. AB - OBJECTIVE: To determine to what degree changes in retinal nerve fiber layer (RNFL) thickness after optic neuritis (ON) correlate with either visual recovery or impairment. METHODS: ON can cause visible defects within the RNFL, which can be quantified using optical coherence tomography (OCT). It may be possible to predict visual recovery by measuring RNFL loss after ON. Fifty-four patients underwent repeated evaluations with optical coherence tomography and standardized ophthalmic testing after ON. Regression analyses were used to determine the relationship between RNFL thickness and visual function. RESULTS: Thinning of the RNFL was seen in the majority of patients (74%), and it tended to occur within 3 to 6 months of ON. The average RNFL value was thinner (p<0.0001) in the affected (78 microm) compared with the unaffected eye (100 microm). Patients with incomplete visual recovery demonstrated greater RNFL loss after ON. Regression analyses demonstrated a threshold of RNFL thickness (75 microm), below which RNFL measurements predicted persistent visual dysfunction. INTERPRETATION: Determination of RNFL thickness may predict visual recovery after ON, and lower RNFL values correlate with impaired visual function. Optical coherence tomography may have a potential role as a surrogate marker for axonal integrity within the optic nerve among patients with ON. PMID- 16718704 TI - Frontotemporal dementia: clinicopathological correlations. AB - OBJECTIVE: Frontotemporal lobar degeneration (FTLD) is characterized by impairments in social, behavioral, and/or language function, but postmortem studies indicate that multiple neuropathological entities lead to FTLD. This study assessed whether specific clinical features predict the underlying pathology. METHODS: A clinicopathological correlation was performed on 90 consecutive patients with a pathological diagnosis of frontotemporal dementia and was compared with an additional 24 cases accrued during the same time period with a clinical diagnosis of FTLD, but with pathology not typically associated with frontotemporal dementia. RESULTS: Postmortem examination showed multiple pathologies including tauopathies (46%), FTLD with ubiquitin-positive inclusions (29%), and Alzheimer's disease (17%). The pathological groups manifested some distinct demographic, clinical, and neuropsychological features, although these attributes showed only a statistical association with the underlying pathology. FTLD with ubiquitin-positive inclusions was more likely to present with both social and language dysfunction, and motor neuron disease was more likely to emerge in these patients. Tauopathies were more commonly associated with an extrapyramidal disorder. Alzheimer's disease was associated with relatively greater deficits in memory and executive function. INTERPRETATION: Clinical and neuropsychological features contribute to delineating the spectrum of pathology underlying a patient diagnosed with FTLD, but biomarkers are needed that, together with the clinical phenotype, can predict the underlying neuropathology. PMID- 16718706 TI - Migraine with aura. a risk factor for unprovoked seizures in children? PMID- 16718707 TI - Ibuprofen and the mouse model of Parkinson's disease. PMID- 16718709 TI - Calpainopathy and eosinophilic myositis. PMID- 16718711 TI - Adhesion of a free-standing newton black film onto a solid substrate. PMID- 16718710 TI - Diet and the risk for Alzheimer's disease. PMID- 16718712 TI - Progressive stabilization of zwitterionic structures in [H(Ser)(2-8)]+ studied by infrared photodissociation spectroscopy. PMID- 16718713 TI - The Heck reaction of electron-rich olefins with regiocontrol by hydrogen-bond donors. PMID- 16718714 TI - Self-supported and clean one-step cathodic coupling of activated olefins with benzyl bromide derivatives in a micro flow reactor. PMID- 16718715 TI - Precision study on capillary electrophoresis methods for metacycline. AB - A CE method for metacycline (MTC) determination was investigated in an inter laboratory experiment. Many problems were encountered in this study, most of which were related to the transfer of the method to different CE equipment. The reported problems could be classified into different categories: problems related to the precision, to the parameters in the protocol, and to the MTC peak shape. As the peak shape problem was partially responsible for the poor precision, a new CE method was developed in order to obtain a good MTC peak shape on all equipment. The precision of this new method for MTC determination was examined in an intermediate precision study, where the influence of the factors "time" and "equipment" was investigated. Although the new method could be transferred to different instruments, the precision remained poor mainly due to the contributions of the between-replicate and the between-injection variances. PMID- 16718716 TI - CE as orthogonal technique to HPLC for alprazolam degradation product identification. AB - The control of degradation products is currently a critical issue to the pharmaceutical industry. A degradation product that appeared in alprazolam tablets during their stability assay, 7-chloro-1-methyl-5-phenyl [1,2,4]triazolo[4,3-a]quinolin-4-amine, also named triazolaminoquinoline, was tested as possible candidate in the HPLC method employed for the study. The impurity showed the same retention time and spectra as the degradation product; but as all these compounds are very closely related, a confirmation with an independent technique was necessary, and CE was chosen for that purpose. Problems related to the adsorption of the analytes to the negatively charged silica surface were solved by employing a new polymeric capillary coating consisting of poly(3-aminopropylmethylsiloxane). The polymer provided EOF towards the anode, and the two compounds were separated in less than 8 min in a 60 cm total-length capillary, 75 microm id capillary with a BGE containing 50 mM phosphate buffer at pH 2.0 with 20% ACN. When the sample containing the degradation product was injected, the presence of triazolaminoquinoline was confirmed. PMID- 16718717 TI - Quantification of ganciclovir in human plasma using capillary electrophoresis. AB - A fast, simple, specific capillary electrophoretic method in the MEKC mode for the quantification of the antiviral drug ganciclovir is described. The separation was obtained using a 50 microm id fused-silica capillary, 60 mM borax buffer (pH 9.25) containing 40 mM SDS using ethenoadenosine as the internal standard. Sample preparation was done by ultrafiltration with a Microcon 30 000 kDa filter. The analytes were detected with UV detector at 254 nm. A sufficient sensitivity was achieved by using a bubble cell capillary. The linear range was from 0.5 to 10 mg/L with a LOQ of 0.5 mg/L. Correlation coefficients were better than 0.999 whereas inter- and intraday precision and accuracy were less than 10.7%. The analysis of patients' samples after administration of ganciclovir indicates that the method is suitable for drug monitoring in the clinic. PMID- 16718718 TI - Optimization of capillary electrophoretic enantioseparation for basic drugs with native beta-CD as a chiral selector. AB - This study presents the advantages of the 20 microm inner diameter (id) capillary for the enantioseparation of ten basic drugs with native beta-CD as the chiral selector. The apparent binding constants of each enantiomeric pair were determined to calculate the optimum beta-CD concentration ([beta-CD]opt) and the optimization was subsequently carried out. Comparison of the 20 microm id with 50 microm id were made in terms of the results obtained in the optimization and detection limits. Applying the optimum conditions for each compound, reproducible results (RSD from 0-3; n>5) were obtained for the 20 microm id capillary. Although the sensitivity is lower in the 20 microm id capillary, the LOD determined using this capillary is still found to be acceptable for the ten basic drugs studied. Enhanced resolution and faster analysis times were the main advantages observed with the use of this capillary in enantioseparation. PMID- 16718719 TI - Capillary electrophoresis and the clinical laboratory. AB - Over the past 15 years, CE as an analytical tool has shown great promise in replacing many conventional clinical laboratory methods, such as electrophoresis and HPLC. CE's appeal was that it was fast, used very small amounts of sample and reagents, was extremely versatile, and was able to separate large and small analytes, whether neutral or charged. Because of this versatility, numerous methods have been developed for analytes that are of clinical interest. Other than molecular diagnostic and forensic laboratories CE has not been able to make a major impact in the United States. In contrast, in Europe and Japan an increasing number of clinical laboratories are using CE. Now that automated multicapillary instruments are commercially available along with cost-effective test kits, CE may yet be accepted as an instrument that will be routinely used in the clinical laboratories. This review will focus on areas where CE has the potential to have the greatest impact on the clinical laboratory. These include analyses of proteins found in serum and urine, hemoglobin (A1c and variants), carbohydrate-deficient transferrin, forensic and therapeutic drug screening, and molecular diagnostics. PMID- 16718721 TI - Lewis acid stabilized OPI3: implications for the nature of free OPI3. AB - While reinvestigating the published synthesis of OPI(3), it became evident from the experiments that phosphoryl triodide may only be formed as an intermediate and that the end products of the reaction of OPCl(3) with LiI are P(V) oxides, PI(3), I(2), and LiCl. This is also in agreement with MP2/TZVPP calculations, which assign Delta(r)H degrees (Delta(r)G degrees ) [Delta(r)G degrees in CHCl(3)] for the disproportionation of OPI(3) as -7 (-18) [-17 kJ mol(-1)] (assuming P(4)O(10) as the P(V) oxide). The first products of this reaction visible in a low-temperature in situ (31)P NMR experiment are P(2)I(4) and PI(3), as well as traces of a compound that may be OPCl(2)I. By contrast, it was possible to prepare and structurally characterize Lewis acid [A] stabilized [A]<- OPX(3) adducts, where [A] is Al(OR(F))(3) for X=Br and Al(OR(F))(2)(mu F)Al(OR(F))(3) for X=I (R(F)=C(CF(3))(3)). These adducts are formed on decomposition of PX(4) (+)[Al(OR(F))(4)](-); high yields of Br(3)PO- >Al(OR(F))(3) (delta((31)P)=-65) were obtained, while I(3)PO-->Al(OR(F))(3) (delta((31)P)=-337) and I(3)PO-->Al(OR(F))(2)(mu-F)Al(OR(F))(3) (delta((31)P)= 332) are only formed as by-products. The main product of the room-temperature decomposition of PI(4) (+)[Al(OR(F))(4)](-) is PI(4) (+)[(R(F)O)(3)Al(mu F)Al(OR(F))(3)](-), which was also characterized by X-ray crystallography and was independently prepared from Ag(+)[(R(F)O)(3)Al(mu-F)Al(OR(F))(3)](-), PI(3), and I(2). PMID- 16718720 TI - Electrophoresis of pharmaceutical proteins: status quo. AB - The biotechnology industry has undergone rapid growth in recent years largely due to the development and success of protein-based therapeutics for a wide range of disorders. Similar to traditional pharmaceuticals, characterization of a therapeutic protein for its physicochemical properties, process monitoring and lot release is crucial. Electrophoresis in the slab-gel format has and continues to be a mainstay of the protein laboratory; and more recently, CE has begun to make significant inroads for protein analysis in industrial settings. This review focuses on the electrophoresis of proteins with an emphasis on protein-based therapeutics in the capillary, slab-gel and to a lesser extent, the microchip format. Reported applications of electrophoresis at several stages of the biopharmaceutical industry covering the period of 2000-2005 will be discussed. PMID- 16718722 TI - Self-assembly of semifluorinated dendrons attached to electron-donor groups mediates their pi-stacking via a helical pyramidal column. AB - Semifluorinated first-generation self-assembling dendrons attached via a flexible spacer to electron-donor molecules induce pi-stacking of the donors in the center of a supramolecular helical pyramidal column. These helical pyramidal columns self-organize in various columnar liquid crystal phases that mediate self processing of large single crystal liquid crystal domains of columns and self repair their intracolumnar structural defects. In addition, all supramolecular columns exhibit a columnar phase at lower temperatures that maintains the helical pyramidal columnar supramolecular structure and displays higher intracolumnar order than that in the liquid crystals phases. The results described here demonstrate the universality of this concept, the power of the fluorous phase or the fluorophobic effect in self-assembly and the unexpected generality of pyramidal liquid crystals. PMID- 16718723 TI - Gold catalysis: phenol synthesis in the presence of functional groups. AB - The effect of different substituents, such as bromo, chloromethyl, hydroxymethyl, formyl, acetyl, carboxy, and acylated hydroxymethyl and ammonium groups, on the furan ring of substrates in gold-catalyzed phenol synthesis has been investigated. The furan ring was also replaced by different heterocycles, such as pyrroles, thiophenes, oxazoles, and a 2,4-dimethoxyphenyl group; gold catalysis then delivered no phenols, but occasionally other products were obtained. [Ru(3)(CO)(12)] also catalyzed the conversion of 1 at a low rate, [Os(3)(CO)(12)] failed as a catalyst, and with [Co(2)(CO)(8)] the alkyne complex 19 can be obtained, it does not lead to any phenol but reacts with norbornene to give the product of a Pauson-Khand reaction. Efforts to prepare vinylidene complexes of 1 provided the only evidence for these species; in the presence of a phosphane ligand with ruthenium an interesting deoxygenation to 22 was observed. The phenol 2 c was converted to the allyl ether, a building block for para-Claisen rearrangements, and to the aryl triflate, a building block for cross-coupling reactions. PMID- 16718724 TI - Aromaticity of giant polycyclic aromatic hydrocarbons with hollow sites: super ring currents in super-rings. AB - We present a systematic theoretical study based on semi-empirical, Hartree-Fock (HF), and density functional theory (DFT) models of a series of polycyclic aromatic hydrocarbons (PAHs) that exhibit hollow sites. In this study we focus particularly on the magnetic criteria of aromaticity, namely (1)H NMR and nucleus independent chemical shifts (NICS), and on their relationships with other electronic properties. The computed shifts and NICS indices indicate that an external magnetic field induces exceptionally strong ring currents in even layered PAH doughnuts, in particular in the layer directly adjacent to the central hole of double-layered compounds. These exceptionally strong ring currents also correlate with particularly small HOMO-LUMO gaps and electronic excitation energies and to abnormally high polarizabilities, indicating in turn that these compounds have a more pronounced metallic character. Comparison is made with further depictions of aromaticity in these systems and in [18] [66]annulene rings by employing topological, structural, and energetic criteria. PMID- 16718725 TI - Chiral phosphane alkenes (PALs): simple synthesis, applications in catalysis, and functional hemilability. AB - A simple synthesis of a chiral phosphane alkene (PAL) involves: 1) palladium catalyzed Suzuki coupling of 10-bromo-5H-dibenzo[a,d]cyclohepten-5-ol (1) with phenylboronic acid to give quantitatively 10-phenyl-5H-dibenzo[a,d]cyclohepten-5 ol (2); 2) reaction of 2 with Ph(2)PCl under acidic conditions to give a racemic mixture of the phosphane oxide (10-phenyl-5H-dibenzo[a,d]cyclohepten-5 yl)diphenylphosphane oxide ((Ph)troppo(Ph), 3), which is separated into enantiomers by using high-pressure liquid chromatography (HPLC) on a chiral column; 3) reduction with trichlorosilane to give the enantiomerically pure phosphanes (R)- and (S)-(10-phenyl-5H-dibenzo[a,d]cyclohepten-5 yl)diphenylphosphane ((Ph)tropp(Ph), 4). This highly rigid, concave-shaped ligand serves as a bidentate ligand in Rh(I) and Ir(I) complexes. Catalysts prepared from [Rh(2)(mu(2)-Cl)(2)(C(2)H(4))(4)] and (S)-4 have allowed the efficient enantioselective 1,4-addition of arylboronic acids to alpha,beta-unsaturated carbonyls (Hayashi-Miyaura reaction) (5-0.1 mol % catalyst, up to 95% ee). The iridium complex (S,S)-[Ir((Ph)tropp(Ph))(2)]OTf ((S,S)-6; OTf=SO(3)CF(3)) has been used as a catalyst in the hydrogenation of various nonfunctionalized and functionalized olefins (turnover frequencies (TOFs) of up to 4000 h(-1)) and moderate enantiomeric excesses have been achieved (up to 67% ee). [Ir((Ph)tropp(Ph))(2)]OTf reversibly takes up three equivalents of H(2). The highly reactive octahedral [Ir(H)(2)(OTf)(CH(2)Cl(2))(H(2)-(Ph)tropp(Ph))(2)] could be isolated and contains two hydrogenated monodentate H(2)-(Ph)tropp(Ph) phosphanes, one CH(2)Cl(2) molecule, one triflate anion, and two hydrides. Based on this structure and extensive NMR spectroscopic studies, a mechanism for the hydrogenation reactions is proposed. PMID- 16718726 TI - Stereoelectronic effects on type I 1,2-dyotropic rearrangements in vicinal dibromides. AB - The effects of different substituents on type I-dyotropic rearrangements of open chain and cyclic 1,2-dibromo hydrocarbons have been studied by means of DFT calculations. The activation energy (E(a)) of this transformation decreases with the pi-donor ability of the substituent attached to the reacting ethylenic system. This is due to donation of electronic density by conjugation or hyperconjugation. This donation leads to longer C--C and C--Br bond lengths in the corresponding four-membered transition states (TSs). Linear relationships between the E(a) and either the sigma(p) Hammett substituent constants and the C- C bond length of the TSs were also found. In all cases, the processes have a high value of synchronicity, which is mainly independent on the substituents. A model based on the second-order perturbational analysis for one ethylene unit with two apical bromine radicals accounts for all the computed results. PMID- 16718727 TI - Ab initio molecular dynamics studies of ionic dissolution and precipitation of sodium chloride and silver chloride in water clusters, NaCl(H2O)n and AgCl(H2O)n, n = 6, 10, and 14. AB - An ab initio molecular dynamics method was used to compare the ionic dissolution of soluble sodium chloride (NaCl) in water clusters with the highly insoluble silver chloride (AgCl). The investigations focused on the solvation structures, dynamics, and energetics of the contact ion pair (CIP) and of the solvent separated ion pair (SSIP) in NaCl(H(2)O)(n) and AgCl(H(2)O)(n) with cluster sizes of n = 6, 10 and 14. We found that the minimum cluster size required to stabilize the SSIP configuration in NaCl(H(2)O)(n) is temperature-dependent. For n = 6, both configurations are present as two distinct local minima on the free-energy profile at 100 K, whereas SSIP is unstable at 300 K. Both configurations, separated by a low barrier (<10 kJ mol(-1)), are identifiable on the free energy profiles of NaCl(H(2)O)(n) for n = 10 and 14 at 300 K, with the Na(+)/Cl(-) pairs being internally solvated in the water cluster and the SSIP configuration being slightly higher in energy (<5 kJ mol(-1)). In agreement with the low bulk solubility of AgCl, no SSIP minimum is observed on the free-energy profiles of finite AgCl(H(2)O)(n) clusters. The AgCl interaction is more covalent in nature, and is less affected by the water solvent. Unlike NaCl, AgCl is mainly solvated on the surface in finite water clusters, and ionic dissolution requires a significant reorganization of the solvent structure. PMID- 16718728 TI - A second metalloid Ga18 cluster and its topological similarity to the high pressure Ga-II modification. AB - The topology of many modifications of elemental gallium is reflected in the large variety of metalloid Ga clusters that have been isolated as intermediates on the way from the metastable molecular GaX species (X=Cl, Br, I) by means of disproportionation to the bulk metal. Herein, we report the synthesis and characterization of the first metalloid cluster anion [Ga(18)(PtBu(2))(10)](3-) with the singular core topology that resembles the gallium high-pressure modification Ga-II. The stabilization of the cluster anion through ion-pair contacts with a chainlike "Li(4)Br(2) backbone" is discussed. Furthermore, the compound is discussed in context of the other metalloid clusters Ga(18)R(8) and Ga(22)R(8) (R=SitBu(3)) and their structural relation to the elemental modifications Ga-III and beta-Ga, respectively. PMID- 16718729 TI - Remarkable interplay of redox states and conformational changes in a sterically crowded, cross-conjugated tetrathiafulvalene vinylog. AB - Derivatives of 9-[2-(1,3-dithiol-2-ylidene)ethylidene]thioxanthene have been synthesized using Horner-Wadsworth-Emmons reactions of (1,3-dithiol-2 yl)phosphonate reagents with thioxanthen-9-ylidene-acetaldehyde (5). Further reactions lead to the sterically crowded cross-conjugated "vinylogous tetrathiafulvalene" derivative 9-[2,3-bis-(4,5-dimethyl-1,3-dithiol-2-ylidene) propylidene]thioxanthene (10). X-ray crystallography, solution electrochemistry, optical spectroscopy, spectroelectrochemistry, and simultaneous electrochemistry and electron paramagnetic resonance spectroscopy, combined with theoretical calculations performed at the B3LYP/6-31G(d) level, elucidate the interplay of the electronic and structural properties in these molecules. For compound 10, multistage redox behavior is observed: the overall electrochemical process can be represented by 10-->10(.+)-->10(2+)-->10(4+) with good reversibility for the 10- >10(.+)-->10(2+) transformations. At the tetracation stage there is the maximum gain in aromaticity at the dithiolium and thioxanthenium rings. Theory predicts that for 10, 10(.+), and 10(2+) the trans isomers are more stable than the cis isomers (by ca. 2-18 kJ mol(-1)), whereas for 10(4+) the cis isomer becomes more stable than the trans isomer (by ca. 25 kJ mol(-1)) [trans and cis refer to the arrangement of the two dithiole moieties with respect to the central ==C(R)- C(H)== fragment]. These data explain the detection in cyclic voltammograms of both trans and cis isomers of 10 and 10(.+) during the reduction of 10(4+) at fast scan rates (>100 mV s(-1)) when the cis-trans isomerization is not completed within the timescale of the experiment. The X-ray structure of the charge transfer complex (CTC) of 10 with 2,4,5,7-tetranitrofluorene-9 dicyanomethylenefluorene (DTeF) [stoichiometry: 10(.+)(DTeF)(2) (.-)2 PhCl] reveals a twisted conformation of 10(.+) (driven by the bulky thioxanthene moiety) and provides a very rare example of segregated stacking of a fluorene acceptor in a CTC. PMID- 16718730 TI - Divergent synthesis of L-sugars and L-iminosugars from D-sugars. AB - An efficient divergent synthesis of L-sugars and L-iminosugars from D-sugars is described. The important intermediate, delta-hydroxyalkoxamate, prepared from D glucono-/galactono-1,5-lactone, was cyclized under Mitsunobu conditions to give the O-cyclized oxime compound and the N-cyclized lactam compound as mixtures. A more detailed investigation revealed that the appropriate protecting groups and solvents controlled the specificity for the O-/N-cyclization of the delta hydroxyalkoxamate. Suitable protection at the 6-position of delta hydroxyalkoxamate, derived from D-glucono-1,5-lactone, afforded the corresponding O-alkylation product alone. Thus we succeeded in applying this to the total synthesis of L-iduronic acid. In contrast, with both TBDMS as the protecting group and RCN as the solvent the efficient conversion of D-glucono/galactono-1,5 lactone into the corresponding L-iminosugars (L-idonolactam and L-altronolactam) was achieved. PMID- 16718731 TI - Steric effects in the uncatalyzed and DMAP-catalyzed acylation of alcohols quantifying the window of opportunity in kinetic resolution experiments. AB - The kinetics of the reaction of several alcohols (benzyl alcohol, ethanol, 1 phenylethanol, cyclohexanol, and 1-methyl-1-phenylethanol) with a selection of anhydrides (acetic anyhydride, propionic anhydride, isobutyric anhydride, isovaleric anhydride, and pivalic anhydride) as catalyzed by 4-(N,N dimethylamino)pyridine (DMAP)/triethyl amine have been studied in CH(2)Cl(2) at 20 degrees C. In all cases the reaction kinetics can be described by rate laws containing a DMAP-catalyzed term and an uncatalyzed (background) term. The rate constants for the background reaction respond sensitively to changes in the steric demand of the alcohol and the anhydride substrates, making the reaction of cyclohexanol with acetic anhydride 526 times faster than the reaction with pivalic anhydride. Steric effects are even larger for the catalyzed reaction and the reactivity difference between acetic and pivalic anhydride exceeds a factor of 8000 for the reaction of cyclohexanol. There is, however, no linear correlation between the steric effects on the catalyzed and the uncatalyzed part. As a consequence there are substrate combinations with dominating catalytic terms (such as the reaction of benzyl alcohol with isobutyric anhydride), while other substrate combinations (such as the reaction of cyclohexanol with pivalic anhydride) are characterized through a dominating background process. The implications of these findings for the kinetic resolution of alcohols are discussed. PMID- 16718732 TI - Clinical proteomics: study of a cryogel. AB - Cryoproteins are proteins precipitating at low temperature. Usually, the precipitate contains immunoglobulins (Igs), and are therefore called cryoglobulins. Very rarely, Igs do not precipitate, but, upon cooling, form a gel. Here, we report a case of cryogel observed in a patient presenting with Waldenstrom's disease. Using proteomic tools, a monoclonal IgM was identified as being the cause of the gel formation. Furthermore, addition of H(2)O before incubation at 4 degrees C demonstrated that the monoclonal IgM was precipitable as a type I cryoglobulin (hypocryoglobulin). PMID- 16718733 TI - Matrix-mediated gene transfer to brain cortex and dorsal root ganglion neurones by retrograde axonal transport after dorsal column lesion. AB - BACKGROUND: In previous studies, we showed that the immobilisation of DNAs encoding basic fibroblast growth factor, neurotrophin-3 and brain-derived neurotrophic factor in a gene-activated matrix (GAM) promotes sustained survival of axotomised retinal ganglion cells after optic nerve injury. Here, we evaluated if the immobilisation of DNAs in a GAM could be an effective approach to deliver genes to axotomised dorsal root ganglion (DRG) neurones after spinal cord injury and if the matrix component of the GAM would modulate the deposition of a dense scar at the injury site. METHODS: We evaluated the expression of the thymidine kinase (TK) reporter gene in brain cortex and DRG after a bilateral T8 dorsal column (DC) lesion using PCR, RT-PCR and in situ hybridisation analyses. Collagen based GAMs were implanted at the lesion site and the cellular response to the GAM was assessed using cell-specific markers. RESULTS: At 1 week post-injury, PCR analyses confirmed that DNATK was retrogradely transported from the DC lesion where the GAM was implanted to the brain cortex and to caudal DRG neurones, and RT-PCR analyses showed expression of mRNATK. At 7 weeks post-injury, DNATK was still be detected in the GAM and DRG. In situ hybridisation localised DNATK and mRNATK within fibroblasts, glia, endothelial and inflammatory cells invading the GAM and in DRG neurones. Interestingly, the presence of a GAM also reduced secondary cavitation and scar deposition at the lesion site. CONCLUSIONS: These results establish that GAMs act as bridging scaffolds in DC lesions limiting cavitation and scarring and delivering genes both locally to injury-reactive cells and distally to the cerebral cortex and to DRG neuronal somata through retrograde axonal transport. PMID- 16718734 TI - Mechanism of plasmid delivery by hydrodynamic tail vein injection. II. Morphological studies. AB - BACKGROUND: The efficient delivery of plasmid DNA (pDNA) to hepatocytes by a hydrodynamic tail vein (HTV) procedure has greatly popularized the use of naked nucleic acids. The hydrodynamic process renders onto the tissue increased physical forces in terms of increased pressures and shear forces that could lead to transient or permanent membrane damage. It can also trigger a series of cellular events to seal or reorganize the stretched membrane. Our goal was to study the uptake mechanism by following the morphological changes in the liver and correlate these with the fate of the injected plasmid DNA. METHODS: We utilized both light microscopic (LM) and electron microscopic (EM) techniques to determine the effect of the HTV procedure on hepatocytes and non-parenchymal cells at various times after injection. The LM studies used paraffin-embedded livers with hematoxylin and eosin (H&E) staining. The immune-EM studies used antibodies labeled with sub-nanometer gold particles followed by silver enhancement to identify the location of injected pDNA at the subcellular level. The level of overall damage to liver cells was estimated based on alanine aminotransferase (ALT) release and clearance. RESULTS: Both the LM and EM results showed the appearance of large vesicles in hepatocytes as early as 5 min post injection. The number of vesicles decreased by 20-60 min. Plasmid DNA molecules often appeared to be associated with or inside such vesicles. DNA could also be detected in the space of Disse, in the cytoplasm and in nuclei. Non-parenchymal cells also contained DNA, but HTV-induced vesicles could not be observed in them. CONCLUSIONS: Our studies suggest an alternative or additional pathway for naked DNA into hepatocytes besides direct entry via membrane pores. It may be difficult to prove which of these pathways lead to gene expression, but the membrane pore hypothesis alone appears insufficient to explain why expression happens preferentially in hepatocytes. Further study is needed to delineate the importance of each of these putative pathways and their interrelationship in enabling oligonucleotide (siRNA) activity and pDNA expression. PMID- 16718735 TI - Azathioprine hepatotoxicity and the protective effect of liquorice and glycyrrhizic acid. AB - This study aimed to evaluate the responses of human hepatocytes to azathioprine hepatotoxicity in comparison with the well-studied azathioprine hepatotoxicity in rat hepatocytes and the effects of protective agents to suppress azathioprine hepatotoxicity. Azathioprine presented its hepatotoxicity at clinically relevant concentrations (lower than 10 microm) in primary rat hepatocytes after 48 h of treatment as shown by a severe decrease in cell viability as well as intracellular GSH depletion. However, primary human hepatocytes exhibited only significant intracellular GSH depletion after treatment with azathioprine at these clinically relevant concentrations, while a reduction in cell viability by 29% was only evidenced after 48 h of treatment with azathioprine at the high concentration of 50 microm. In addition, a monolayer culture of primary rat hepatocytes was used as an in vitro model to examine the protective effects of antihepatotoxic drugs including glutathione (GSH), N-acetylcysteine (NAC, a GSH precursor), liquorice and glycyrrhizic acid (GA), a major bioactive component of liquorice, against hepatotoxicity of 1 microm azathioprine. It was found that both liquorice and GA showed substantial protection according to assays of cell viability and intracellular GSH, while neither GSH nor NAC had such a protective function. Similarly, GA protected human hepatocytes from intracellular GSH depletion on exposure to 1 microm azathioprine. These results implied that GA or liquorice could be considered as potent protection agents against azathioprine hepatotoxicity. PMID- 16718736 TI - The protein fraction of Phyllanthus niruri plays a protective role against acetaminophen induced hepatic disorder via its antioxidant properties. AB - The aim of this study was to investigate the hepatoprotective action of the protein fraction of Phyllanthus niruri against acetaminophen (APAP) hepatotoxicity. The partially purified protein fraction of P. niruri was injected intraperitoneally in mice either prior to (preventive) or after the induction of toxicity (curative). Levels of different liver marker enzymes in serum and different antioxidant enzymes, as well as lipid peroxidation in total liver homogenates were measured in normal, control (toxicity induced) and P. niruri protein fraction-treated mice. P. niruri significantly reduced the elevated glutamate pyruvate transaminase (GPT) and alkaline phosphatase (ALP) levels in the sera of toxicity induced mice, compared with the control group. Lipid peroxidation levels were also reduced in mice treated with P. niruri protein fraction compared with the APAP treated control group. Among the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST) levels were restored to almost normal levels compared with the control group. P. niruri treatment also enhanced reduced hepatic glutathione (GSH) levels caused by APAP administration. The results demonstrated that the protein fraction of P. niruri protected liver tissues against oxidative stress in mice, probably acting by increasing antioxidative defense. PMID- 16718737 TI - Cytotoxicity in vitro of naphthazarin derivatives from Onosma arenaria. AB - The cytotoxicity of naphthazarin derivatives isolated from the roots of Onosma arenaria on human cervix adenocarcinoma cells (HeLa) and leukaemia K562 cells, as well on non-malignant peripheral blood mononuclear cells (PBMC) was studied. The results show that beta-hydroxyisovalerylalkannin, acetylalkannin and the pigment fraction exhibited high cytotoxicity in vitro against the tested cell lines, as well the healthy PBMC before or after activation with phytohaemagglutinin. PMID- 16718738 TI - Pesticide exposure assessment in rice paddies in Europe: a comparative study of existing mathematical models. AB - A comparative test was undertaken in order to identify the potential of existing mathematical models, including the rice water quality (RICEWQ) 1.6.4v model, the pesticide concentration in paddy field (PCPF-1) model and the surface water and groundwater (SWAGW) model, for calculating pesticide dissipation and exposure in rice paddies in Europe. Previous versions of RICEWQ and PCPF-1 models had been validated under European and Japanese conditions respectively, unlike the SWAGW model which was only recently developed as a tier-2 modelling tool. Two datasets, derived from field dissipation studies undertaken in northern Italy with the herbicides cinosulfuron and pretilachlor, were used for the modelling exercise. All models were parameterized according to field experimentations, as far as possible, considering their individual deficiencies. Models were not calibrated against field data in order to remove bias in the comparison of the results. RICEWQ 1.6.4v provided the highest agreement between measured and predicted pesticide concentrations in both paddy water and paddy soil, with modelling efficiency (EF) values ranging from 0.78 to 0.93. PCPF-1 simulated well the dissipation of herbicides in paddy water, but significantly underestimated the concentrations of pretilachlor, a chemical with high affinity for soil sorption, in paddy soil. SWAGW simulated relatively well the dissipation of both herbicides in paddy water, and especially pretilachlor, but failed to predict closely the pesticide dissipation in paddy soil. Both RICEWQ and SWAGW provided low groundwater (GW) predicted environmental concentrations (PECs), suggesting a low risk of GW contamination for the two herbicides. Overall, this modelling exercise suggested that RICEWQ 1.6.4v is currently the most reliable model for higher-tier exposure assessment in rice paddies in Europe. PCPF-1 and SWAGW showed promising results, but further adjustments are required before these models can be considered as strong candidates for inclusion in the higher-tier pesticide regulatory scheme. PMID- 16718739 TI - Detection of resistance to acetolactate synthase inhibitors in weeds with emphasis on DNA-based techniques: a review. AB - Resistance to herbicides inhibiting acetolactate synthase (ALS) has been increasing at a faster rate than in any other herbicide group. The great majority of these cases are due to various single-nucleotide polymorphisms in the ALS gene endowing target site resistance. Many diagnostic techniques have been devised in order to confirm resistance and help producers to adopt the best management strategies. Recent advances in DNA technologies coupled with the knowledge of sequence information have allowed the development of accurate and rapid diagnostic tests. While whole plant-based diagnostic techniques such as seedling bioassays or enzyme-based in vitro bioassays provide accurate results, they tend to be labour- and/or space-intensive and will only respond to the particular herbicides tested, making resolution of cross-resistance patterns more difficult. Successful DNA-based diagnosis of ALS inhibitor resistance has been achieved with three main techniques, (1) restriction fragment length polymorphism, (2) polymerase chain reaction amplification of specific alleles and (3) denaturing high-performance liquid chromatography. All DNA-based techniques are relatively rapid and provide clear identification of the mutations causing resistance. Resistance based on non-target mechanisms is not identified by these DNA-based methods; however, given the prevalence of target site-based ALS inhibitor resistance, this is a minor inconvenience. PMID- 16718740 TI - Variation of Musca domestica L. acetylcholinesterase in Danish housefly populations. AB - Anti-cholinesterase resistance is in many cases caused by modified acetylcholinesterase (MACE). A comparison was made of toxicological data and AChE activity gathered from 21 field populations and nine laboratory strains of houseflies, Musca domestica L., to elucidate the best way of generating data to provide advice for management strategies and gathering information for resistance risk assessment on the organophosphates azamethiphos and dimethoate and the carbamate methomyl, which have been the primary insecticides used against adult houseflies in Denmark. Cluster analysis was performed and > 2000 houseflies were assigned to one of three phenotypes based on total acetylcholinesterase activity as well as inhibition by azamethiphos, methomyl or omethoate. A cluster, i.e. a phenotype, with high total AChE activity and high sensitivity to azamethiphos and less sensitivity to inhibition by methomyl and omethoate was shown to be linked to methomyl resistance. It was not possible to define any clusters that could be linked to azamethiphos or dimethoate resistance. The five mutations V180L, G262A, G262V, F327Y and G365A causing anticholinesterase resistance in houseflies were all identified in the Danish housefly strains. The data are very heterogeneous, and a correlation of molecular genetic background and resistance of phenotypes is not obvious with the available data. PMID- 16718743 TI - Cuticular uptake of xenobiotics into living plants. Part 2: influence of the xenobiotic dose on the uptake of bentazone, epoxiconazole and pyraclostrobin, applied in the presence of various surfactants, into Chenopodium album, Sinapis alba and Triticum aestivum leaves. AB - This study has determined the uptake of three pesticides, applied as commercial or model formulations in the presence of a wide range of surfactants, into the leaves of three plant species (bentazone into Chenopodium album L. and Sinapis alba L., epoxiconazole and pyraclostrobin into Triticum aestivum L.). The results have confirmed previous findings that the initial dose (nmol mm(-2)) of xenobiotic applied to plant foliage is a strong, positive determinant of uptake. This held true for all the pesticide formulations studied, although surfactant concentration was found to have an effect. The lower surfactant concentrations studied showed an inferior relationship between the amount of xenobiotic applied and uptake. High molecular mass surfactants also produced much lower uptake than expected from the dose uptake equations in specific situations. PMID- 16718744 TI - Rodenticide grain bait ingredient acceptance by Norway rats (Rattus norvegicus), California ground squirrels (Spermophilus beecheyi) and pocket gophers (Thomomys bottae). AB - Vertebrate pest control in California is often accomplished through the use of rodenticide grain baits. These grain baits are composed of steam-rolled oats (SRO), a toxicant, an indicator dye and an oil combination. A series of tests were performed to determine the effects of various dye and oil formulations on acceptance of grain bait by Norway rats [Rattus norvegicus (Berk)], California ground squirrels [Spermophilus beecheyi (Richardson)] and pocket gophers (Thomomys bottae Eyd & Gerv). Seven different dyes, four oil formulations and clean (untreated) oats were tested for acceptance. The addition of the selected oils and dyes to grain resulted in no significant differences in consumption. This indicates that there is a wide variety of dyes that could be used in the formulation of rodenticides. These alternatives could aid in proper pesticide use, the deterrence of bait consumption by birds and possibly in ingredient adhesion to the finished bait. PMID- 16718745 TI - Captan and fenitrothion dissipation in field-treated cauliflowers and effect of household processing. AB - Field trial studies have been performed with captan and fenitrothion on cauliflower to propose maximum residue limits and to study the dissipation of the pesticides. Residue levels have been determined at different times following good laboratory practice using gas chromatography with mass spectrometric detection. The behaviour of residue levels of these compounds after household processing has been analysed using gas chromatography with electron capture detection. Seven days after treatment, residue levels of captan could be detected, but not of fenitrothion. The half-lives of dissipation for captan and fenitrothion were calculated as 0.9 and 1.8 days respectively. Washing did not significantly affect the residual amounts of captan and fenitrothion observed in raw vegetables; however, after cooking, captan had degraded completely, whereas residue levels of fenitrothion were not modified significantly. PMID- 16718746 TI - Gene expression analysis of dendritic/Langerhans cells and Langerhans cell histiocytosis. AB - Langerhans cell histiocytosis (LCH) is a neoplastic disorder that results in clonal proliferation of cells with a Langerhans cell (LC) phenotype. The pathogenesis of LCH is still poorly understood. In the present study, serial analysis of gene expression (SAGE) was applied to LCs generated from umbilical cord blood CD34+ progenitor cells to identify LC-specific genes and the expression of these genes in LCH was investigated. Besides the expression of several genes known to be highly expressed in LCs and LCH such as CD1a, LYZ, and CD207, high expression of genes not previously reported to be expressed in LCs, such as GSN, MMP12, CCL17, and CCL22, was also identified. Further analysis of these genes by quantitative RT-PCR revealed high expression of FSCN1 and GSN in all 12 LCH cases analysed; of CD207, MMP12, CCL22, and CD1a in the majority of these cases; and CCL17 in three of the 12 cases. Immunohistochemistry confirmed protein expression in the majority of cases. The expression of MMP12 was most abundant in multi-system LCH, which is the LCH type with the worst prognosis. This suggests that expression of MMP12 may play a role in the progression of LCH. These data reveal new insight into the pathology of LCH and provide new starting points for further investigation of this clonal proliferative disorder. PMID- 16718747 TI - Endoscopic therapies of gastroesophageal reflux disease. AB - The high prevalence of gastroesophageal reflux disease (GERD) in Western societies has accelerated the need for new modalities of treatment. Currently, medical and surgical therapies are widely accepted among patients and physicians. New potent antisecretory drugs and the development of minimally invasive surgery for the management of GERD are at present the pivotal and largely accepted approaches to treatment. The minimally invasive treatment revolution, however, has stimulated several new endoscopic techniques for GERD. Up to now, the data is limited and further studies are necessary to compare the advantages and disadvantages of the various endoscopic techniques to medical and laparoscopic management of GERD. New journal articles and abstracts are continuously being published. The Food and Drug Administration has approved 3 modalities, thus gastroenterologists and surgeons are beginning to apply these techniques. Further trials and device refinements will assist clinicians. This article will present an overview of the various techniques that are currently on study. This review will report the efficacy and durability of various endoscopic therapies for gastroesophageal reflux disease (GERD). The potential for widespread use of these techniques will also be discussed. Articles and abstracts published in English on this topic were retrieved from Pubmed. Due to limited number of studies and remarkable differences between various trials, strict criteria were not used for the pooled data presented, however, an effort was made to avoid bias by including only studies that used off-PPI scoring as baseline and intent to treat. PMID- 16718748 TI - Reassessment of functional dyspepsia: a topic review. AB - Dyspepsia itself is not a diagnosis but stands for a constellation of symptoms referable to the upper gastrointestinal tract. It consists of a variable combination of symptoms including abdominal pain or discomfort, postprandial fullness, abdominal bloating, early satiety, nausea, vomiting, heartburn and acid regurgitation. Patients with heartburn and acid regurgitation invariably have gastroesophageal reflux disease and should be distinguished from those with dyspepsia. There is a substantial group of patients who do not have a definite structural or biochemical cause for their symptoms and are considered to be suffering from functional dyspepsia (FD). Gastrointestinal motor abnormalities, altered visceral sensation, dysfunctional central nervous system-enteral nervous system (CNS-ENS) integration and psychosocial factors have all being identified as important pathophysiological correlates. It can be considered as a biopsychosocial disorder with dysregulation of the brain-gut axis being central in origin of disease. FD can be categorized into different subgroups based on the predominant single symptom identified by the patient. This subgroup classification can assist us in deciding the appropriate symptomatic treatment for the patient. PMID- 16718749 TI - Epidemiology of functional dyspepsia: a global perspective. AB - Dyspepsia refers to group of upper gastrointestinal symptoms that occur commonly in adults. Dyspepsia is known to result from organic causes, but the majority of patients suffer from non-ulcer or functional dyspepsia. Epidemiological data from population-based studies of various geographical locations have been reviewed, as they provide more realistic information. Population-based studies on true functional dyspepsia (FD) are few, due to the logistic difficulties of excluding structural disease in large numbers of people. Globally, the prevalence of uninvestigated dyspepsia (UD) varies between 7%-45%, depending on definition used and geographical location, whilst the prevalence of FD has been noted to vary between 11%-29.2%. Risk factors for FD have been shown to include females and underlying psychological disturbances, whilst environmental/ lifestyle habits such as poor socio-economic status, smoking, increased caffeine intake and ingestion of non-steroidal anti-inflammatory drugs appear to be more relevant to UD. It is clear that dyspepsia and FD in particular are common conditions globally, affecting most populations, regardless of location. PMID- 16718750 TI - Subtypes of functional dyspepsia. AB - Functional dyspepsia is a common clinical condition characterised by chronic or recurrent upper abdominal pain or discomfort commonly associated with a variety of associated gastrointestinal symptoms and a normal endoscopy. To standardise research-based approaches, an initial categorisation of into sub groups was agreed to, based on clusters of symptoms. However the early expectation that these subgroups would be associated with distinct pathophysiologies amenable to specific therapy has not been realised. A classification based on the most troublesome symptom has been suggested but the utility of this is also unclear. More recent data suggest that some of the pathophysiologic dysfunctions may be associated with specific symptoms and so provide a better tool for grouping patients. But this approach remains incomplete as current insights into the pathogenesis are still too limited for this to be satisfactory. In conclusion, no classification provides for an adequate treatment-based approach to the syndrome of functional dyspepsia. As a consequence treatment remains largely empiric. PMID- 16718751 TI - Functional dyspepsia: the role of visceral hypersensitivity in its pathogenesis. AB - Functional, or non-ulcer, dyspepsia (FD) is one of the most common reasons for referral to gastroenterologists. It is associated with significant morbidity and impaired quality of life. Many authorities believe that functional dyspepsia and irritable bowel syndrome represent part of the spectrum of the same disease process. The pathophysiology of FD remains unclear but several theories have been proposed including visceral hypersensitivity, gastric motor dysfunction, Helicobacter pylori infection and psychosocial factors. In this review, we look at the evidence, to date, for the role of visceral hypersensitivity in the aetiology of FD. PMID- 16718752 TI - Role of Helicobacter pylori in functional dyspepsia. AB - The aetiology of dyspepsia is unknown in the majority of patients. Helicobacter pylori (H pylori) is the cause in a subset of patients. A non invasive test to assess the presence of H pylori is recommended in the management of patients under the age of 50 presenting to a family practitioner with dyspepsia. A urea breath test or a stool antigen test are the most reliable non invasive tests. Eradication of H pylori will reduce the risk to the patient with dyspepsia of developing a peptic ulcer, reduce the complication rate if prescribed non-steroid anti-inflammatory drugs and later reduce the risk of gastric cancer. The recommended treatment for non ulcer dyspepsia associated with a H pylori infection should be a 10-d course of treatment with a PPI and two antibiotics. Treatment efficacy should be assessed four weeks after completing treatment with a urea breath test or a stool antigen test. PMID- 16718753 TI - Central serotonergic and noradrenergic receptors in functional dyspepsia. AB - Functional dyspepsia is a symptom complex characterised by upper abdominal discomfort or pain, early satiety, motor abnormalities, abdominal bloating and nausea in the absence of organic disease. The central nervous system plays an important role in the conducting and processing of visceral signals. Alterations in brain processing of pain, perception and affective responses may be key factors in the pathogenesis of functional dyspepsia. Central serotonergic and noradrenergic receptor systems are involved in the processing of motor, sensory and secretory activities of the gastrointestinal tract. Visceral hypersensitivity is currently regarded as the mechanism responsible for both motor alterations and abdominal pain in functional dyspepsia. Some studies suggest that there are alterations in central serotonergic and noradrenergic systems which may partially explain some of the symptoms of functional dyspepsia. Alterations in the autonomic nervous system may be implicated in the motor abnormalities and increases in visceral sensitivity in these patients. Noradrenaline is the main neurotransmitter in the sympathetic nervous system and again alterations in the functioning of this system may lead to changes in motor function. Functional dyspepsia causes considerable burden on the patient and society. The pathophysiology of functional dyspepsia is not fully understood but alterations in central processing by the serotonergic and noradrenergic systems may provide plausible explanations for at least some of the symptoms and offer possible treatment targets for the future. PMID- 16718754 TI - Cholecystokinin hyperresponsiveness in functional dyspepsia. AB - Functional dyspepsia (FD) is a common disorder of yet uncertain etiology. Dyspeptic symptoms are usually meal related and suggest an association to gastrointestinal (GI) sensorimotor dysfunction. Cholecystokinin (CCK) is an established brain-gut peptide that plays an important regulatory role in gastrointestinal function. It inhibits gastric motility and emptying via a capsaicin sensitive vagal pathway. The effects on emptying are via its action on the proximal stomach and pylorus. CCK is also involved in the regulation of food intake. It is released in the gut in response to a meal and acts via vagal afferents to induce satiety. Furthermore CCK has also been shown to be involved in the pathogenesis of panic disorder, anxiety and pain. Other neurotransmitters such as serotonin and noradrenaline may be implicated with CCK in the coordination of GI activity. In addition, intravenous administration of CCK has been observed to reproduce the symptoms in FD and this effect can be blocked both by atropine and loxiglumide (CCK-A antagonist). It is possible that an altered response to CCK may be responsible for the commonly observed gastric sensorimotor dysfunction, which may then be associated with the genesis of dyspeptic symptoms. PMID- 16718755 TI - Drug treatment of functional dyspepsia. AB - Symptomatic improvement of patients with functional dyspepsia after drug therapy is often incomplete and obtained in not more than 60% of patients. This is likely because functional dyspepsia is a heterogeneous disease. Although great advance has been achieved with the consensus definitions of the Rome I and II criteria, there are still some aspects about the definition of functional dyspepsia that require clarification. The Rome criteria explicitly recognise that epigastric pain or discomfort must be the predominant complaint in patients labelled as suffering from functional dyspepsia. However, this strict definition can create problems in the daily primary care clinical practice, where the patient with functional dyspepsia presents with multiple symptoms. Before starting drug therapy it is recommended to provide the patient with an explanation of the disease process and reassurance. A thorough physical examination and judicious use of laboratory data and endoscopy are also indicated. In general, the approach to treat patients with functional dyspepsia based on their main symptom is practical and effective. Generally, patients should be treated with acid suppressive therapy using proton-pump inhibitors if the predominant symptoms are epigastric pain or gastroesophageal reflux symptoms. Although the role of Helicobacter pylori (H pylori) in functional dyspepsia continues to be a matter of debate, recent data indicate that there is modest but clear benefit of eradication of H pylori in patients with functional dyspepsia. In addition, H pylori is a gastric carcinogen and if found it should be eliminated. Although there are no specific diets for patients with FD, it may be helpful to guide the patients on healthy exercise and eating habits. PMID- 16718756 TI - Functional dyspepsia: are psychosocial factors of relevance? AB - The pathogenesis of Functional Dyspepsia (FD) remains unclear, appears diverse and is thus inadequately understood. Akin to other functional gastrointestinal disorders, research has demonstrated an association between this common diagnosis and psychosocial factors and psychiatric morbidity. Conceptualising the relevance of these factors within the syndrome of FD requires application of the biopsychosocial model of disease. Using this paradigm, dysregulation of the reciprocal communication between the brain and the gut is central to symptom generation, interpretation and exacerbation. Appreciation and understanding of the neurobiological correlates of various psychological states is also relevant. The view that psychosocial factors exert their influence in FD predominantly through motivation of health care seeking also persists. This appears too one dimensional an assertion in light of the evidence available supporting a more intrinsic aetiological link. Evolving understanding of pathogenic mechanisms and the heterogeneous nature of the syndrome will facilitate effective management. Co morbid psychiatric illness warrants treatment with conventional therapies. Acknowledging the relevance of psychosocial variables in FD, the degree of which is subject to variation, has implications for assessment and management. Available evidence suggests psychological therapies may benefit FD patients particularly those with chronic symptoms. The rationale for use of psychotropic medications in FD is apparent but the evidence base to support the use of antidepressant pharmacotherapy is to date limited. PMID- 16718757 TI - Functional dyspepsia and irritable bowel syndrome, are they different entities and does it matter? AB - A high prevalence of overlap between functional dyspepsia and irritable bowel syndrome has been consistently and universally reported. Recent studies demonstrating shared common pathophysiological disturbances including delayed gastric emptying and visceral hypersensitivity involving more than one region, suggest that these patients have a generalised rather than regional, disorder of the gut. Furthermore, a study of the natural history of dyspepsia suggests that with time, a substantial proportion will evolve into IBS. The recognition of IBS in dyspeptic patients has potentially profound therapeutic importance. It could help to reduce the risk of unnecessary cholecystectomy in IBS patients. The ability to appreciate the extent of involvement could allow us to address the disturbances more comprehensively, and thereby achieve greater patient satisfaction with their treatment. PMID- 16718758 TI - Latest insights into the effects of Helicobacter pylori infection on gastric carcinogenesis. AB - There appears to be the strong association between Helicobacter pylori (H pylori) and gastric cancer. We reviewed the latest evidences about the effects of H pylori infection on gastric carcinogenesis, classified into epidemiology, dynamics of gastric mucosal changes, DNA damages, virulence factors, host factors, and source of gastric malignancy. Through the considerable progress made in research into virulence factors resulting from differences between H pylori strains, such as cagA positivity, as well as into host factors, such as gene polymorphisms, a diverse spectrum of H pylori-associated diseases, including gastric cancer, is beginning to lend itself to elucidation. The impact of the novel hypothesis advanced by Houghton et al proposing bone-marrow derived stem cells (BMDC) as a potential source of gastric malignancy on evolving research remains to be seen with interest. Further progress in research into H pylori eradication as a viable prophylaxis of gastric cancer, as well as into the mechanisms of gastric carcinogenesis, is to be eagerly awaited for the current year and beyond. PMID- 16718760 TI - Is interferon-beta an alternative treatment for chronic hepatitis C? AB - The treatment of chronic hepatitis C (CHC) is still far from optimal, particularly for those subpopulations that do not respond to the standard combination therapy with Interferon-alpha (IFNalpha) plus ribavirin. Although in some cases the use of higher doses or longer treatment periods may be effective, these approaches are generally associated with a higher incidence of adverse events, which may either lead to a reduction in patient compliance or require drug withdrawal. IFNbeta could represent an interesting alternative for treating CHC patients. Controversial data about IFNbeta efficacy in CHC exist, the main reason being that many results stem from pilot studies with small cohorts of patients. However, promising results have been obtained in some subgroups of patients that fail to respond to IFNalpha. Additionally, the good tolerability of IFNbeta represents an important advantage of the drug. The rates of dropouts in controlled clinical trials, as well as the need for dose reductions or treatment discontinuation are very low. It might be worth assessing the value of IFNbeta plus ribavirin in randomized studies with a larger cohort of patients, not eligible or not tolerating standard therapy, and for non-responders. PMID- 16718761 TI - Splenic vasculopathy in portal hypertension patients. AB - AIM: To investigate the interaction between portal hypertension, splanchnic hyperdynamic circulation and splanchnic vasculopathy by observing splenic arterial and venous pathological changes and the ro1e of extra-cellular matrix in the pathogenesis of portal hypertensive vasculopathy by measuring the expression of type I and type III procollagen mRNA in splenic venous walls of portal hypertensive patients. METHODS: Morphological changes of splenic arteries and veins taken from portal hypertensive patients (n=20) and normal controls (n=10) were observed under optical and electron microscope. Total RNA was extracted and the expression of type I and type III procollagen mRNA in splenic venous walls of portal hypertensive patients (n=20) was semi-quantitatively detected using reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Under optical microscope, splenic arterial intima was destroyed and internal elastic membrane and medial elastic fibers of the splenic arterial walls were degenerated and broken. Splenic venous intima became remarkably thick. Endothelia1 cells were not intact with formation of mural thrombus. The tunica media became thickened significantly due to hypertrophy of smooth muscles. Fibers and connective tissues were increased obviously. Under electron microscope, smooth muscle cells of the splenic arteries were degenerated and necrotized. Phenotypes of smooth muscle cells changed from constrictive into synthetic type. Red blood cells and platelets accumulated around the damaged endothelial cells. Synthetic smooth muscle cells were predominant in splenic veins and their cytoplasma had plentiful rough endoplasmic reticulum ribosomes and Golgi bodies. Along the vascular wall, a lot of collagen fibers were deposited, the intima was damaged and blood components accumulated. There was no significant difference in the expression of type I procollagen mRNA in splenic venous wall between the patients with portal hypertension and those without portal hypertension (P>0.05), but the expression of type III procoagen mRNA was significantly stronger in the patients with portal hypertension than in those without portal hypertension (P<0.01). CONCLUSION: Type III procollagen and collagen might be important extra-cellular matrix resulting in neointimal formation and vascular remodeling in the pathogenesis of portal hypertensive vasculopathy. The pathological changes in splenic arteries and veins exist in portal hypertension patients. There might be an interaction between portal hypertension, splanchnic hyperdynamic circulation and splanchnic vasculopathy. PMID- 16718759 TI - A concomitant review of the effects of diabetes mellitus and hypothyroidism in wound healing. AB - This paper reviews the negative impact of diabetes mellitus or hypothyroidism on wound healing, both in experimental and clinical settings. Since both are metabolic disorders of great clinical importance, special attention is given, not only to their pathophysiology, but also to their biochemical and histological effects on tissue integrity and regeneration. Also, special focus is awarded on wound healing of the gastrointestinal tract, i.e. in intestinal anastomosis, and how these disorders can lead to wound dehiscence. Since diabetes mellitus and hypothyroidism can coexist in clinical settings, more research must be directed on their influence on wound healing, considering them as one clinical entity. PMID- 16718762 TI - Pharmacokinetics and bioequivalence of ranitidine and bismuth derived from two compound preparations. AB - AIM: To evaluate the bioequivalence of ranitidine and bismuth derived from two compound preparations. METHODS: The bioavailability was measured in 20 healthy male Chinese volunteers following a single oral dose (equivalent to 200 mg of ranitidine and 220 mg of bismuth) of the test or reference products in the fasting state. Then blood samples were collected for 24 h. Plasma concentrations of ranitidine and bismuth were analyzed by high-performance liquid chromatography and inductively coupled plasma-mass spectrometry (ICP-MS), respectively. The non compartmental method was used for pharmacokinetic analysis. Log-transformed C(max), AUC( (0-t) ) and AUC( (0-infinity) ) were tested for bioequivalence using ANOVA and Schuirmann two-one sided t-test. T(max) was analyzed by Wilcoxon's test. RESULTS: Various pharmacokinetic parameters of ranitidine derived from the two compound preparations, including C(max), AUC( (0-t)), AUC( (0-infinity)), T(max) and T((1/2)), were nearly consistent with previous observations. These parameters derived from test and reference drug were as follows: C(max) (0.67 +/- 0.21 vs 0.68 +/-0.22 mg/L), AUC( (0-t) ) (3.1 +/- 0.6 vs 3.0 +/- 0.7 mg/L per hour), AUC( (0-infinity) ) (3.3 +/- 0.6 vs 3.2 +/- 0.8 mg/L per hour), T(max) (2.3 +/- 0.9 vs 2.1 +/- 0.9 h) and T((1/2)) (2.8 +/- 0.3 vs 3.1 +/- 0.4 h). In addition, double-peak absorption profiles of ranitidine were found in some Chinese volunteers. For bismuth, those parameters derived from test and reference drug were as follows: C(max) (11.80 +/- 7.36 vs 11.40 +/- 6.55 microg/L), AUC( (0 t) ) (46.65 +/- 16.97 vs 47.03 +/- 21.49 microg/L per hour), T(max) (0.50 +/- 0.20 vs 0.50 +/- 0.20 h) and T((1/2)) (10.2 +/- 2.3 vs 13.0 +/- 6.9 h). Ninety percent of confidence intervals for the test/reference ratio of C(max), AUC( (0 t) ) and AUC( (0-infinity) ) derived from both ranitidine and bismuth were found within the bioequivalence acceptable range of 80%-125%. No significant difference was found in T(max) derived from both ranitidine and bismuth. CONCLUSION: The two compound preparations are bioequivalent and may be prescribed interchangeably. PMID- 16718763 TI - Modulatory effects of Azadirachta indica on benzo(a)pyrene-induced forestomach tumorigenesis in mice. AB - AIM: To evaluate the chemopreventive effects of aqueous Azadirachta indica (A indica) leaf extract (AAILE) against benzo(a)pyrene [B(a)P]-induced forestomach tumorigenesis in Balb/c mice. METHODS: Female Balb/c mice were divided into four groups of 10-12 animals each. For induction of forestomach tumors, starting from d 14 of the experi-ment, mice of B(a)P and B(a)P+A indica groups were given intra gastric instillations of B(a)P (40 mg/kg), twice a week for four weeks. Mice of A indica and B(a)P+A indica groups were orally administered with AAILE (100 mg/kg), two weeks prior to B(a)P instillations till the end of the experiment. After 22 wk of the first B(a)P instillation, mice were sacrificed and the forestomachs were analyzed for development of tumors, scanning electron microscopy (SEM) and histopathology. RESULTS: Tumor incidence was observed to be 100% in mice that received only B(a)P. However, treatment with AAILE reduced the tumor incidence by 58.4% as observed in mice of B(a)P+A indica group when compared to that of B(a)P group. Similarly, the tumor burden and multiplicity were seen to decrease by 87.3% and 69.6% respectively in mice of B(a)P+A indica group when compared to those of B(a)P group. Scanning electron microscopy analysis showed that AAILE treatment itself did not cause any abnormalities on the surface architecture of forestomach epithelium. In tumorous forestomach, surface disruption was observed. Over the forestomach tumors of B(a)P group of mice certain rounded structures were seen in addition to closely placed tongue-shaped squamous cells. Interestingly, these rounded structures were not observed in B(a)P + A indica group of mice. Histopathalogically, the tumors were identical and diagnosed to be papillomas. Mice from control and A indica groups of mice did not develop any forestomach tumors and showed normal histo-architecture. CONCLUSION: The present data suggest that A indica exerts chemopreventive effects against B(a)P-induced forestomach tumors in murine model. Because of lack of toxicity and ubiquitous bioavailability, A indica may play a promising role in future drug discovery and development as far as chemoprevention of cancer is concerned. PMID- 16718764 TI - Efficacy and safety of ecabet sodium on functional dyspepsia: a prospective, double-blinded, randomized, multi-center controlled trial. AB - AIM: To compare ecabet sodium and cimetidine in relieving symptoms of functional dyspepsia. METHODS: We performed a multi-center, prospective, randomized, double blinded controlled trial to compare the clinical efficacy of ecabet sodium and cimetidine in patients with functional dyspepsia. Two-hundred and seventy-two patients with dyspeptic symptoms fulfilling the Rome-II criteria were enrolled from 7 centers. In the study group (115 patients), 1.5 g ecabet sodium was given twice a day. In the control group (121 patients), 400 mg cimetidine was given twice a day. Symptoms and parameters of quality of life were analyzed at baseline, 3, 14, and 28 d after initiating the treatment. RESULTS: Two-hundred and thirty-six patients completed the clinical trial. After 4 wk of treatment, the rates of improvement in patients with dyspeptic symptoms were not different between two groups (77.4% in the ecabet group and 79.3% in the cimetidine group, respectively, P > 0.05). Likewise, the rates of symptomatic improvement were not different at 3 d and 14 d. The parameters of quality of life did not change significantly during the study period in both groups. There was no clinically significant adverse event in both groups. CONCLUSION: In patients with functional dyspepsia, ecabet sodium has similar clinical efficacy with cimetidine. PMID- 16718766 TI - Effect of somatostatin analogue octreotide injected into the third cerebral ventricle on pentagastrin-induced gastric acid secretion in rats. AB - AIM: To investigate the effect of long-lasting somatostatin analogue octreotide (Oct) injected into the third cerebral ventricle (TCV) on gastric acid secretion in rats. METHODS: TCVs were cannulated in male Wistar rats anesthetized with sodium pentobarbital. One week later acute gastric lumen perfusion was carried out and gastric acid was continuously washed with 37 degree Celsius saline by a perfusion pump. Gastric perfusion samples were collected every 10 min and titrated by 0.01 moL/L NaOH to neutral. On the basis of subcutaneous (sc) injection of pentagastrin (G-5, 160 microg/kg), Oct (0.025 microg, 0.05 microg, 0.1 microg, n=12 in each group) or vehicle (pyrogen-free physiological saline, n=10) was injected into the TCV. Before and after the TCV injection, 1 h total acid output (TAO) was determined and experimental data were expressed in change rate (%) of TAO. RESULTS: Oct (0.025, 0.05 and 0.1 microg) injected into the TCV resulted in change rate of 1.56% (P>0.05), 20.21% (P<0.01) and 37.82% of TAO (P<0.001), respectively. Moreover, comparison in change rate of TAO among these 3 doses showed P<0.05 between 0.025 microg and 0.05 microg, P<0.01 between 0.025 microg and 0.1 microg, and P<0.05 between 0.05 microg and 0.1 microg. However, sc injection of 0.05 microg Oct had no effect on G-5 stimulated gastric acid secretion. CONCLUSION: Octreotide injected into the third cerebral ventricle inhibits gastrin-induced gastric acid secretion in a dose-dependent manner. PMID- 16718765 TI - Expression of angiostatin cDNA in human gallbladder carcinoma cell line GBC-SD and its effect on endothelial proliferation and growth. AB - AIM: To explore the influence of angiostatin up-regulation on the biologic behavior of gallbladder carcinoma cells in vitro and in vitro, and the potential value of angiostatin gene therapy for gallbladder carcinoma. METHODS: A eukaryotic expression vector of pcDNA3.1(+) containing murine angiostatin was constructed and identified by restriction endonuclease digestion and sequencing. The recombinant vector pcDNA3.1-angiostatin was transfected into human gallbladder carcinoma cell line GBC-SD with Lipofectamine 2000, and paralleled with the vector and mock control. The resistant clone was screened by G418 filtration. Angiostatin transcription and protein expression were examined by RT PCR, immunofluorescence and Western-blot. The supernatant was collected to treat endothelial cells. Cell proliferation and growth in vitro were observed under microscope. RESULTS: Murine angiostatin cDNA was successfully cloned into the eukaryotic expression vector pcDNA3.1 (+). After 14 d of transfection and selection with G418, macroscopic resistant cell cloning was formed in the experimental group transfected with pcDNA 3.1(+)-angiostatin and vector control. But untreated cells died in the mock control. Angiostatin was detected by RT-PCR and protein expression was detected in the experimental group by immunofluorescence and Western-blot. Cell proliferation and growth in vitro in the three groups were observed respectively under microscope. No significant difference was observed in the growth speed of GBC-SD cells between groups that were transfected with and without angiostatin. After treatment with supernatant, significant differences were observed in endothelial cell (ECV-304) growth in vitro. The cell proliferation and growth were inhibited. CONCLUSION: Angiostatin does not directly inhibit human gallbladder carcinoma cell proliferation and growth in vitro, but the secretion of angiostatin inhabits endothelial cell proliferation and growth. PMID- 16718767 TI - Clinico-pathological aspects of colorectal serrated adenomas. AB - AIM: To study the association of colorectal serrated adenomas (SAs) with invasive carcinoma, local recurrence, synchronicity and metachronicity of lesions. METHODS: A total of 4,536 polyps from 1,096 patients over an eight-year period (1987-1995) were retrospectively examined. Adenomas showing at least 50% of serrated architecture were called SAs by three reviewing pathologists. RESULTS: Ninety-one (2%) of all polyps were called SAs, which were found in 46 patients. Invasive carcinomas were seen in 3 out of 46 (6.4%) patients, of whom one was a case of familial adenomatous polyposis (FAP). A male preponderance was noted and features of a mild degree of dysplasia were seen in majority (n=75, 83%) of serrated adenomas. Follow-up ranged 1-12 years with a mean time of 5.75 years. Recurrences of SAs were seen in 3 (6.4%) cases, synchronous SAs in 16 (34.8%) cases and metachronous SAs in 9 (19.6%) cases. CONCLUSION: Invasive carcinoma arising in serrated adenoma is rare, accounting for 2 (4.3%) cases studied in this series. PMID- 16718768 TI - Platelet-activating factor in cirrhotic liver and hepatocellular carcinoma. AB - AIM: Platelet-activating factor (PAF) is a pro-inflammatory and angiogenic lipid mediator. Here we aimed to investigate levels of PAF, lyso-PAF (the PAF precursor), phospholipase A(2) (PLA(2), the enzymatic activity generating lyso PAF), acetylhydrolase activity (AHA, the PAF degrading enzyme) and PAF receptor (PAF-R) transcripts in cirrhotic liver and hepatocellular carcinoma (HCC). METHODS: Twenty-nine patients with HCC were enrolled in this study. Cirrhosis was present in fourteen patients and seven had no liver disease. Tissue PAF levels were investigated by a platelet-aggregation assay. Lyso-PAF was assessed after its chemical acetylation into PAF. AHA was determined by degradation of [(3)H] PAF. PLA(2) levels were assessed by EIA. PAF-R transcripts were investigated using RT-PCR. RESULTS: Elevated amounts of PAF and PAF-R transcripts 1 (leukocyte type) were found in cirrhotic tissues as compared with non-cirrhotic ones. Higher amounts of PAF and PAF-R transcripts 1 and 2 (tissue-type) were found in HCC tissues as compared with non-tumor tissues. PLA(2), lyso-PAF and AHA levels were not changed in cirrhotic tissues and HCC. CONCLUSION: While the role of PAF is currently unknown in liver physiology, this study suggests its potential involvement in the inflammatory network found in the cirrhotic liver and in the angiogenic response during HCC. PMID- 16718769 TI - Determination of anti-endomysium IgA antibodies in the diagnosis of celiac disease: comparison of a novel ELISA-based assay with conventional immunofluorescence. AB - AIM: To evaluate the novel anti-endomysium (anti-EMA) detection based on ELISA. METHODS: Anti-EMA IgA was measured by a novel ELISA in 196 patients with gastrointestinal symptoms and suspected mal-absorption. Data were compared with those obtained by the conventional IF test. RESULTS: A good concordance of 98% was found between these two assays. In sera of 161 patients (82%) both assays tested negative whereas in sera of 31 patients (16%) both assays tested positive for the presence of anti-EMA antibodies. Discrepancies between EMA-ELISA and EMA immunofluorescence (IF) were found in only 4 patients (2%). CONCLUSION: This ELISA can replace IF for the detection of anti-EMA antibodies and provide clinicians with an excellent tool to screen for celiac disease in patients with gastrointestinal complaints. PMID- 16718770 TI - Establishment of model of visceral pain due to colorectal distension and its behavioral assessment in rats. AB - AIM: To establish a visceral pain model via colorectal distension (CRD) and to evaluate the efficiency of behavioral responses of CRD by measuring the score of abdominal withdrawal reflex (AWR) in rats. METHODS: Thirty-eight male SD rats weighing 180-240 g were used to establish the visceral pain model. The rat was inserted intra-anally with a 7 cm long flexible latex balloon under ether anesthesia, and colorectal distensions by inflating the balloon with air were made 30 min after recovering from the anesthesia. Five AWR scores (AWR0 to AWR4) were used to assess the intensity of noxious visceral stimuli. It was regarded as the threshold of the minimal pressure (kPa). For abdominal flatting was induced by colorectal distension. RESULTS: A vigorous AWR to distension of the descending colon and rectum was found in 100% of the awake rats tested. The higher the pressure of distension, the higher the score of AWR. The distension pressures of 0, 2.00, 3.33, 5.33 and 8.00 kPa produced different AWR scores (P<0.05). The pain threshold of AWR was constant for up to 80 min after the initial windup (first 1 3 distensions), the mean threshold was 3.69+/-0.35 kPa. Systemic administration of morphine sulfate elevated the threshold of visceral pain in a dose-dependent and naloxone reversible manner. CONCLUSION: Scoring the AWR during colorectal distensions can assess the intensity of noxious visceral stimulus. Flatting of abdomen (AWR 3) to CRD as the visceral pain threshold is clear, constant and reliable. This pain model and its behavioral assessment are good for research on visceral pain and analgesics. PMID- 16718771 TI - Generation and characterization of a transgenic mouse model for pancreatic cancer. AB - AIM: To generate a SV40Tag transgenic tumor animal model and to study the mechanism underlying tumorigenesis. METHODS: A mammary gland expression vector containing SV40Tag DNA was generated. Transgene fragments were microinjeted into fertilized eggs of FVB mice. The genetically manipulated embryos were transferred into the oviducts of pseudo-pregnant female mice. PCR and Northern blot analysis were used for genotype analysis of F1 and F2 mice. Transgene expression was detected by RT-PCR and immunohistochemistry. RESULTS: SV40Tag gene was detected in two lines of transgenic mice. One of them delivered the transgene to F1 and a tumor was found in the pancreas of these mice. RT-PCR and immunohistochemistry showed that SV40Tag gene was expressed in the tumor. Pathological characterization of the transgenic mice demonstrated that the tumor belonged to pancreatic cystic neoplasm. CONCLUSION: SV40Tag transgenic mouse model can be successfully established. The transgenic mice develop a pancreatic tumor, which can be used for investigation of the molecular mechanism of tumorigenesis in vivo. PMID- 16718772 TI - Analysis of variabilities of serum proteomic spectra in patients with gastric cancer before and after operation. AB - AIM: To study the variabilities of serum proteomic spectra in patients with gastric cancer before and after operation in order to detect the specific protein markers that can be used for quick diagnosis of gastric cancer. METHODS: Proteomic spectra of 46 serum samples from patients with gastric cancer before and after operation and 40 from normal individuals were generated by IMAC-Cu protein chip and surface-enhanced laser desorption/ ionization time of flight mass spectrometry. RESULTS: Fourteen differentially expressed proteins in serum were screened by analysis of proteomic spectra of preoperative patients and normal individuals. We obtained 4 proteins (heat shock protein 27, glucose regulated protein, prohibitin, protein disulfide isomerase A3) making up marker pattern which was able to class the patient-team and normal-team. These marker patterns yielded 95.7% sensitivity and 92.5% specificity, respectively. The proteins over-expressed in serum of preoperative patients were obviously down regulated. CONCLUSION: Specific protein markers of gastric cancer can be used for the quick diagnosis of gastric cancer and judgment of prognosis. SELDI-TOF-MS is a useful tool for the detection and identification of new protein markers in serum. PMID- 16718773 TI - Long-term survival of a case with multiple liver metastases from duodenal gastrointestinal stromal tumor drastically reduced by the treatment with imatinib and hepatectomy. AB - Constitutive activation of KIT receptor tyrosine kinase is a critical factor in the pathogenesis of gastrointestinal stromal tumors (GISTs). But there is little information on whether combination of imatinib mesylate (IM) and surgical treatment can prolong survival in the cases with unresectable multiple liver metastases. We report a case of postoperative recurrence of GIST treated by the tyrosine kinase inhibitor IM and surgical treatment. The initial complete response (CR) to treatment continued for 18 mo, but single liver metastasis showed regrowth in the left hepatic lobe during IM treatment. After partial resection of the recurrent tumor, postoperative course was uneventful and the patient has survived without recurrence for 24 mo. Currently, imatinib is the first-line therapy for non-resectable GISTs, but a single agent therapy often leads to tumor resistance. Even if tolerance to imatinib occurs, a combination of imatinib and surgical treatment can prolong survival in some cases as reported here. However, further studies on a large number of cases of recurrent GIST are necessary to evaluate the effectiveness of IM treatment combined with surgery. PMID- 16718774 TI - Post-traumatic hepatic artery pseudo-aneurysm combined with subphrenic liver abscess treated with embolization. AB - A 23-year-old man with post-traumatic hepatic artery pseudo-aneurysm and subphrenic liver abscess was admitted. He underwent coil embolization of hepatic artery pseudo-aneurysm. The pseudo-aneurysm was successfully obstructed and subphrenic liver abscess was controlled. Super-selective trans-catheter coil embolization may represent an effective treatment for hepatic artery pseudo aneurysm combined with subphrenic liver abscess in the absence of other therapeutic alternatives. PMID- 16718775 TI - Current and future applications of magnetic resonance imaging and spectroscopy of the brain in hepatic encephalopathy. AB - Hepatic encephalopathy (HE) is a common neuro-psychiatric abnormality, which complicates the course of patients with liver disease and results from hepatocellular failure and/or portosystemic shunting. The manifestations of HE are widely variable and involve a spectrum from mild subclinical disturbance to deep coma. Research interest has focused on the role of circulating gut-derived toxins, particularly ammonia, the development of brain swelling and changes in cerebral neurotransmitter systems that lead to global CNS depression and disordered function. Until recently the direct investigation of cerebral function has been difficult in man. However, new magnetic resonance imaging (MRI) techniques provide a non-invasive means of assessment of changes in brain volume (coregistered MRI) and impaired brain function (fMRI), while proton magnetic resonance spectroscopy (1H MRS) detects changes in brain biochemistry, including direct measurement of cerebral osmolytes, such as myoinositol, glutamate and glutamine which govern processes intrinsic to cellular homeostasis, including the accumulation of intracellular water. The concentrations of these intracellular osmolytes alter with hyperammonaemia. MRS-detected metabolite abnormalities correlate with the severity of neuropsychiatric impairment and since MR spectra return towards normal after treatment, the technique may be of use in objective patient monitoring and in assessing the effectiveness of various treatment regimens. PMID- 16718776 TI - Cellular and molecular aspects of gastric cancer. AB - Gastric cancer remains a global killer with a shifting burden from the developed to the developing world. The cancer develops along a multistage process that is defined by distinct histological and pathophysiological phases. Several genetic and epigenetic alterations mediate the transition from one stage to another and these include mutations in oncogenes, tumour suppressor genes and cell cycle and mismatch repair genes. The most significant advance in the fight against gastric cancer came with the recognition of the role of Helicobacter pylori (H pylori) as the most important acquired aetiological agent for this cancer. Recent work has focussed on elucidating the complex host/microbial interactions that underlie the neoplastic process. There is now considerable insight into the pathogenesis of this cancer and the prospect of preventing and eradicating the disease has become a reality. Perhaps more importantly, the study of H pylori-induced gastric carcinogenesis offers a paradigm for understanding more complex human cancers. In this review, we examine the molecular and cellular events that underlie H pylori induced gastric cancer. PMID- 16718777 TI - Helicobacter pylori infection causes gastric cancer? A review of the epidemiological, meta-analytic, and experimental evidence. AB - Since the discovery of Campylobacter-like organisms Helicobacter pylori (H pylori) more than two decades ago the possibility of a relationship with gastric cancer has been postulated, tested and supposedly proven. There have been numerous human studies of various designs from many countries around the world. Several meta-analyses have been published and more recently a small number of experimental animal studies were reported looking at the association between H pylori infection and gastric cancer. Over the years, the human epidemiological studies have produced conflicting results; the meta-analyses have as one would expect produced similar pooled estimates; while the early experimental animal studies require replication. The exact mechanisms by which H pylori might cause gastric cancer are still under investigation and remain to be elucidated. PMID- 16718778 TI - Selective inhibition of cell growth by activin in SNU-16 cells. AB - AIM: To investigate whether activin regulates the cell proliferation of human gastric cancer cell line SNU-16 through the mRNA changes in activin receptors, Smads and p21(CIP1/WAF1). METHODS: The human gastric cancer cell lines were cultured, RNAs were purified, and RT-PCRs were carried out with specifically designed primer for each gene. Among them, the two cell lines SNU-5 and SNU-16 were cultured with activin A for 24, 48 and 72 h. The cell proliferation was measured by MTT assay. For SNU-16, changes in ActRIA, ActRIB, ActRIIA, ActRIIB, Smad2, Smad4, Smad7, and p21(CIP1/WAF1) mRNAs were detected with RT-PCR after the cells were cultured with activin A for 24, 48 and 72 h. RESULTS: The proliferation of SNU-16 cells was down regulated by activin A whereas other cells showed no change. Basal level of inhibin/activin subunits, activin receptors, Smads, and p21(CIP1/WAF1) except for activin betaB mRNAs was observed to have differential expression patterns in the human gastric cancer cell lines, AGS, KATO III, SNU-1, SNU-5, SNU-16, SNU-484, SNU-601, SNU-638, SNU-668, and SNU-719. Interestingly, significantly higher expressions of ActR IIA and IIB mRNAs were observed in SNU-16 cells when compared to other cells. After activin treatment, ActR IA, IB, and IIA mRNA levels were decreased whereas ActR IIB mRNA level increased in SNU-16 cells. Smad4 mRNA increased for up to 48 h whereas Smad7 mRNA increased sharply at 24 h and returned to the initial level at 48 h in SNU-16 cells. In addition, expression of the p21(CIP1/WAF1), the mitotic inhibitor, peaked at 72 h after activin treatment in SNU-16 cells. CONCLUSION: Our results suggest that inhibition of cell growth by activin is regulated by the negative feedback effect of Smad7 on the activin signaling pathway, and is mediated through p21(CIP1/WAF1) activation in SNU-16 cells. PMID- 16718779 TI - Inhibition of hepatitis B virus DNA replicative intermediate forms by recombinant interferon-gamma. AB - AIM: To evaluate the in vitro anti-HBV activity of recombinant human IFN-gamma, alone and in combination with lamivudine. METHODS: A recombinant baculovirus HBV/HepG2 culture system was developed which could support productive HBV infection in vitro. Expression of HBsAg and HBeAg in infected HepG2 culture medium was detected by commercial enzyme immunoassays. HBV DNA replication intermediates were detected in infected cells by Southern hybridization and viral DNA load was determined by dot hybridization. RESULTS: IFN-gamma at 0.1 to 5 microg/L efficiently down regulated HBsAg expression in transduced HepG2 cells. At 5 microg/L, IFN-gamma also suppressed HBV DNA replication in these cells. While treatment with a combination of lamivudine and IFN-gamma showed no additive effect, sequential treatment first with lamivudine and then IFN-gamma was found to be promising. In this culture system the best HBV suppression was observed with a pulse of 2 micromol/L lamivudine for two days, followed by 1 microg/L IFN gamma for another four days. Compared to treatment with lamivudine alone, the sequential use of 0.2 micromol/L lamivudine for two days, followed by 5 microg/L IFN-gamma for six days showed a 72% reduction in HBV cccDNA pool. CONCLUSION: This in vitro study warrants further evaluation of a combination of IFN-gamma and lamivudine, especially in IFN-alpha non-responder chronic hepatitis B patients. A reduced duration of lamivudine treatment would also restrict the emergence of drug-resistant HBV mutants. PMID- 16718780 TI - T cell immune response is correlated with fibrosis and inflammatory activity in hepatitis B cirrhotics. AB - AIM: To explore the relationship among interferon-gamma (IFN-gamma) activity, fibrogenesis, T cell immune responses and hepatic inflammatory activity. METHODS: Peripheral blood samples from a total of 43 hepatitis B cirrhotic patients (LC) and 19 healthy controls (NC) were collected to measure their serum levels of IFN gamma, interleukin-2 (IL-2), soluble interleukin-2 receptor (sIL-2R), interleukin 10 (IL-10) and three serological markers of fibrosis including hyaluronic acid (HA), procollagen type III peptide (PIIIP), and type IV collagen were measured using a double antibody sandwich ELISA. Also, serum total bilirubin (TB) and alanine aminotransferase (ALT) were measured by routine measures. RESULTS: The concentrations of serological markers of fibrosis in patients with active cirrhosis (ALC) were significantly higher than those in stationary liver cirrhosis (SLC) or NC groups. The levels of serological markers in HBeAg-positive patients were significantly higher than those in HBeAg-negative patients. In SLC and ALC patients, a negative linear correlation was found between IFN-gamma levels and the serological markers of fibrosis. IFN-gamma and IL-2 levels in the ALC group were significantly higher than those in the SLC and NC groups, but the statistical difference was not significant between the latter two. In contrast, IL-10 levels in the SLC group were significantly higher than that in the NC group, but no significant difference was found between SLC and ALC groups. The sIL-2R level was elevated gradually in all these groups, and the differences were significant. Positive linear correlations were seen between IFN-gamma activity and ALT levels (r = 0.339, P < 0.05), and IL-2 activity and TB levels (r = 0.517, P < 0.05). sIL-2R expression was positively correlated with both ALT and TB levels (r = 0.324, 0.455, P < 0.05), whereas there was no statistically significant correlation between IL-10 expression and serum ALT and TB levels (r = -0.102, -0.093, P > 0.05). Finally, there was a positive correlation between IFN gamma and IL-2 levels. CONCLUSION: T cell immune responses are correlated with fibrosis and hepatic inflammatory activity and may play an important role in liver cirrhosis. PMID- 16718781 TI - Image cytometric DNA analysis of mucosal biopsies in patients with primary achalasia. AB - AIM: To determine DNA aneuploidy in mucosal biopsies of achalasia patients for subsequent rapid diagnosis. METHODS: Biopsies from the middle third of the esophagus were obtained in 15 patients with achalasia. Immunohistochemical staining was carried out with monoclonal antibodies MIB-1 for Ki67 and PAb 1801 for p53, in addition to the conventional histologic examination for dysplasia. Nuclei of fresh biopsy material were enzymatically and mechanically isolated, and the DNA content was determined with image cytometry after Feulgen staining. DNA grading of malignancy was assessed according to Boecking to determine the variability of DNA values noted around the normal diploid peak. Further indices measured included the aneuploid rate, and the 5c-, 7c- and 9c-exceeding rate. RESULTS: The histological examination did not demonstrate dysplasia; while MIB-1 (basal) showed a positive reaction in 8/15 achalasia specimens, p53 was negative in all specimens. Image cytometric DNA analysis detected aneuploidy in 4/15 (26.7%) specimens. Samples from 15 patients with squamous cell carcinoma as well as specimens obtained exclusively 2 cm proximal to the tumor served as reference tests. All carcinomas (15/15) as well as 9 of the peritumoral samples (9/15) were aneuploid. The comparison of biopsies from achalasia patients with peritumoral and carcinoma specimens revealed statistically significant differences regarding the aneuploid rate (diploid: P < 0.0001; tetraploid: P = 0.001), grading of malignancy according to Boecking (P < 0.0001) and the 5c- (P < 0.0001), 7c- (P < 0.0001), and 9c- (P = 0.0001) exceeding rate with progredient DNA alterations in the respective order. CONCLUSION: The finding that DNA aneuploidy was identified by image cytometry in esophageal specimens of patients with achalasia, which may be due to specific chromosomal alterations presenting as precancerous lesions in 27% of patients, leads us to conclude that image cytometry represents a valuable screening tool. PMID- 16718782 TI - Gamma-aminobutyric acid secreted from islet beta-cells modulates exocrine secretion in rat pancreas. AB - AIM: To investigate the role of endogenous gamma-amino-butyric acid (GABA) in pancreatic exocrine secretion. METHODS: The isolated, vascularly perfused rat pancreas was employed in this study to eliminate the possible influences of extrinsic nerves and hormones. Cholecystokinin (CCK; 10 pmol/L) was intra arterially given to stimulate exocrine secretion of the pancreas. RESULTS: Glutamine, a major precursor of GABA, which was given intra-arterially at concentrations of 1, 4 and 10 mmol/L, dose-dependently elevated the CCK stimulated secretions of fluid and amylase in the normal pancreas. Bicuculline (10 micromol/L), a GABA(A) receptor antagonist, blocked the enhancing effect of glutamine (4 mmol/L) on the CCK-stimulated exocrine secretions. Glutamine, at concentrations of 1, 4 and 10 mmol/L, dose-dependently increased the GABA concentration in portal effluent of the normal pancreas. The effects of glutamine on the CCK-stimulated exocrine secretion as well as the GABA secretion were markedly reduced in the streptozotocin-treated pancreas. CONCLUSION: GABA could be secreted from beta-cells into the islet-acinar portal system after administration of glutainine, and could enhance the CCK-stimulated exocrine secretion through GABA(A) receptors. Thus, GABA in islet beta-cells is a hormone modulating pancreatic exocrine secretion. PMID- 16718783 TI - Gastro-protective action of lafutidine mediated by capsaicin-sensitive afferent neurons without interaction with TRPV1 and involvement of endogenous prostaglandins. AB - AIM: Lafutidine, a histamine H2 receptor antagonist, exhibits gastro-protective action mediated by capsaicin-sensitive afferent neurons (CSN). We compared the effect between lafutidine and capsaicin, with respect to the interaction with endogenous prostaglandins (PG), nitric oxide (NO) and the afferent neurons, including transient receptor potential vanilloid subtype 1 (TRPV1). METHODS: Male SD rats and C57BL/6 mice, both wild-type and prostacyclin IP receptor knockout animals, were used after 18 h of fasting. Gastric lesions were induced by the po administration of HCl/ethanol (60% in 150 mmol/L HCl) in a volume of 1 mL for rats or 0.3 mL for mice. RESULTS: Both lafutidine and capsaicin (1-10 mg/kg, po) afforded dose-dependent protection against HCl/ethanol in rats and mice. The effects were attenuated by both the ablation of CSN and pretreatment with N(G) nitro-L-arginine methyl ester, yet only the effect of capsaicin was mitigated by prior administration of capsazepine, the TRPV1 antagonist, as well as indomethacin. Lafutidine protected the stomach against HCl/ethanol in IP receptor knockout mice, similar to wild-type animals, while capsaicin failed to afford protection in the animals lacking IP receptors. Neither of these agents affected the mucosal PGE2 or 6-keto PGF(1alpha) contents in rat stomachs. Capsaicin evoked an increase in [Ca2+]i in rat TRPV1-transfected HEK293 cells while lafutidine did not. CONCLUSION: These results suggest that although both lafutidine and capsaicin exhibit gastro-protective action mediated by CSN, the mode of their effects differs regarding the dependency on endogenous PGs/IP receptors and TRPV1. It is assumed that lafutidine interacts with CSN at yet unidentified sites other than TRPV1. PMID- 16718784 TI - Helicobacter species ribosomal DNA in the pancreas, stomach and duodenum of pancreatic cancer patients. AB - AIM: To determine whether gastric and enteric Helicobacter species are associated with pancreatic cancer. METHODS: Patients with exocrine pancreatic cancer (n = 40), neuroendocrine cancer (n = 14), multiple endocrine neoplasia type 1 (n = 8), and chronic pancreatitis (n = 5) were studied. Other benign pancreatic diseases (n = 10) and specimens of normal pancreas (n = 7) were included as controls. Pancreatic tissue specimens were analyzed by Helicobacter-specific PCR-assay and products were characterized by denaturing gradient electrophoresis and DNA sequencing. From a subset of the pancreatic cancer patients, gastric and/or duodenal tissue as well as gallbladder and ductus choledochus tissue were analyzed. Gallbladder and choledochus samples were included as controls. Stomach and duodenum samples were investigated to analyze whether a gastric helicobacter might disseminate to the pancreas in pancreatic cancer patients. Pancreatic specimens were analyzed by Bacteroides-specific PCR for detecting the translocation of indigenous gut microbes to the diseased pancreas. RESULTS: Helicobacter DNA was detected in pancreas (tumor and/or surrounding tissue) of 75% of patients with exocrine cancer, 57% of patients with neuroendocrine cancer, 38% of patients with multiple endocrine neoplasia, and 60% of patients with chronic pancreatitis. All samples from other benign pancreatic diseases and normal pancreas were negative. Thirty-three percent of the patients were helicobacter-positive in gastroduodenal specimens. Surprisingly, H. bilis was identified in 60% of the positive gastroduodenal samples. All gallbladder and ductus choledochus specimens were negative for helicobacter. Bacteroides PCR assay was negative for all pancreatic samples. CONCLUSION: Helicobacter DNA commonly detected in pancreatic cancer suggests a possible role of the emerging pathogens in the development of chronic pancreatitis and pancreatic cancer. PMID- 16718785 TI - Correlation between TIMP-1 expression and liver fibrosis in two rat liver fibrosis models. AB - AIM: To evaluate serum TIMP-1 level and the correlation between TIMP-1 expression and liver fibrosis in immune-induced and CCL4-induced liver fibrosis models in rats. METHODS: Immune-induced and CCL4-induced liver fibrosis models were established by dexamethasone (0.01 mg) and CCL4 respectively. Serum TIMP-1 level was detected with ELISA, while histopathological grade of liver biopsy was evaluated. Spearman rank-correlation test was used to analyse the difference of the correlation between the TIMP-1 expression and hepatic fibrosis in the two fibrosis models. Furthermore, in situ hybridization was used to determine the expression difference of TIMP-1 mRNA in the two models. RESULTS: Positive correlation existed between serum TIMP-1 level of immune induced group and the histopathological stages of fibrosis liver of corresponding rats (Spearman rank correlation test, r(s) = 0.812, P < 0.05), and the positive in situ hybridization signal of TIMP-1 mRNA was strong. In CCL4-induced liver fibrosis model, the correlation between the serum TIMP-1 level and the severity of hepatic fibrosis was not statistically significant(Spearman rank-correlation test, r(s) = 0.229, P > 0.05). And compared with immune-induced model, the positive in situ hybridization signal of TIMP-1 mRNA was weaker, while the expression variation was higher in hepatic fibrosis of the same severity. CONCLUSION: The correlations between TIMP-1 expression and liver fibrosis in two rat liver fibrosis models are different. In immune-induced model, serum TIMP-1 level could reflect the severity of liver fibrosis, while in CCL4-induced model, the correlation between the serum TIMP-1 level and the severity of hepatic fibrosis was not statistically significant. PMID- 16718786 TI - An efficient method of sorting liver stem cells by using immuno-magnetic microbeads. AB - AIM: To develop a method to isolate liver stem cells fast and efficiently. METHODS: Fetal mouse liver cells were characterized by cell surface antigens (c Kit and CD45/TER119) using flow cytometry. The candidate liver stem cells were sorted by using immuno-magnetic microbeads and identified by clone-forming culture, RT-PCR and immunofluorescence assays. RESULTS: The c-Kit-(CD45/TER119)- cell population with 97.9% of purity were purified by immuno-magnetic microbeads at one time. The yield of this separation was about 6% of the total sorting cells and the cell viability was above 98%. When cultured in vitro these cells had high clone-forming and self-renewing ability and expressed markers of hepatocytes and bile duct cells. Functionally mature hepatocytes were observed after 21 d of culture. CONCLUSION: This method offers an excellent tool for the enrichment of liver stem cells with high purity and viability, which could be used for further studies. It is fast, efficient, simple and not expensive. PMID- 16718787 TI - Effect of recombinant human interleukin-11 on expressions of interleukin-11 receptor alpha-chain and glycoprotein 130 in intestinal epithelium cell line-6 after neutron irradiation. AB - AIM: To explore the effect of recombinant human interleukin-11 (rhIL-11) on the expressions of interleukin-11 receptor alpha-chain (IL-11Ralpha) and an additional signal transducer glycoprotein 130 (gp130) in intestinal epithelium cell line-6 (IEC-6) after neutron irradiation. METHODS: Cultured IEC-6 cells were exposed to 4.0Gy neutron and treated with 100 ng/mL rhIL-11 12 h prior to or immediately after irradiation. The apoptosis and necrosis rates and expressions of IL-11Ralpha and gp130 were observed by flow cytometry, immunohistochemistry, Western blot and image analysis. RESULTS: The apoptosis rate of IEC-6 cells was increased by irradiation at 6 h (P < 0.01), IL-11 stimulation resulted in a decreased apoptosis rate in irradiated IEC-6 cells (P < 0.05). In normal control IEC-6 cells, intense immunoreactivity of IL-11Ralpha was located within the cell membrane and cytoplasm. The level of IL-11Ralpha expression significantly decreased at 6 h after irradiation (P < 0.01) and restored at 24 h after irradiation. In IEC-6 cells treated with both radiation and rhIL-11, the level of IL-11Ralpha expression was higher than that of irradiated cells (P < 0.05). When it came to gp130 protein, it was located in the cytoplasm of IEC-6 cells. After irradiation, we found a progressive decrease in the expression of gp130 protein (P < 0.05) in 48 h post-radiation, while in rhIL-11-stimulated cells, it came back to normal level at 24 h after irradiation and decreased at 48 h, but was still higher than that of only irradiated cells (P < 0.05). CONCLUSION: rhIL-11 can protect IEC-6 cells from neutron irradiation. The protective effect of rhIL 11 might be connected with its ability to up-regulate the expressions of specific ligand-binding subunit IL-11Ralpha and signal-transducing subunit gp130. PMID- 16718788 TI - Effect of early administration of exogenous basic fibroblast growth factor on acute edematous pancreatitis in rats. AB - AIM: To observe the therapeutic effect of early administration of exogenous Basic fibroblast growth factor (bFGF) on acute edematous pancreatitis (AEP) in rats. METHODS: Thirty male Sprague-Dawley rats were randomly divided into three (n = 10): normal control group (group I), AEP group (group II) and AEP with bFGF treatment group (group III). AEP was induced by subcutaneous injection of cerulein (5.5 microg/kg and 7.5 microg/kg) at 1 h interval into rats of groups II and III. Three hours after induction of AEP, 100 microg/kg bFGF was administrated intraperitoneally for 1 h to group III rats. For test of DNA synthesis in acinar cells, 5-bromo-2'-deoxyuridine (BrdU) labeling solution was intraperitoneally injected into the rats of groups II and III 24 h after bFGF treatment. The changes in serum amylase, lipase, pancreatic tissue wet/dry ratio were detected. RESULTS: In bFGF treatment group, there was a significant decrease in the volume of serum amylase, lipase and the pancreatic wet/dry weight ratio(1383.0+/-94.6 U/L, 194.0+/-43.6 U/L, 4.32+/-0.32) compared to AEP group (3464+/-223.7 U/L, 456+/-68.7 U/L, 6.89+/-0.47) (P < 0.01), and no significant difference was found between bFGF treatment and control group (1289+/-94.0 U/L, 171+/-23.4 U/L, 4.12+/ 0.26, P > 0.05). The inflammatory changes such as interstitial edema, polymorphonuclear neutrophils (PMNs) and vacuolization were significantly ameliorated compared to AEP group (P < 0.01). A small number of BrdU-labeled nuclei were observed in acinar cells of AEP rats (1.8+/-0.3 nuclei/microscopic field, n = 10) while diffuse BrdU-labeled nuclei were found in bFGF-treated rats (18.9+/-1.4 nuclei/microscopic field, n = 10) (P < 0.01). Immunohistochemical study showed increased DNA synthesis in pancreatic acinar cells. CONCLUSION: Early administration of exogenous bFGF has significant therapeutic effect on cerulein-induced acute edematous pancreatitis in rats. Its mechanism is related to the amelioration of inflammation and facilitation of pancreatic regeneration. PMID- 16718789 TI - Hepatitis B virus x gene and cyanobacterial toxins promote aflatoxin B1-induced hepatotumorigenesis in mice. AB - AIM: To assess the combinative role of aflatoxin B1(AFB1), cyanobacterial toxins (cyanotoxins), and hepatitis B virus (HBV) x gene in hepatotumorigenicity. METHODS: One-week-old animals carrying HBV x gene and their wild-type littermates were intraperitoneally (ip) injected with either single-dose AFB1 [6 mg/kg body weight (bw)], repeated-dose cyanotoxins (microcystin-LR or nodularin, 10 microg/kg bw once a week for 15 wk), DMSO (vehicle control) alone, or AFB1 followed by cyanotoxins a week later, and were sacrificed at 24 and 52 wk post treatment. RESULTS: AFB1 induced liver tumors in 13 of 29 (44.8%) transgenic mice at 52 wk post-treatment, significantly more frequent than in wild-type mice (13.3%). This significant difference was not shown in the 24-wk study. Compared with AFB1 exposure alone, MC-LR and nodularin yielded approximately 3-fold and 6 fold increases in the incidence of AFB(1)-induced liver tumors in wild-type animals at 24 wk, respectively. HBV x gene did not further elevate the risk associated with co-exposure to AFB1 and cyanotoxins. With the exception of an MC LR-dosed wild-type mouse, no liver tumor was observed in mice treated with cyanotoxins alone at 24 wk. Neither DMSO-treated transgenic mice nor their wild type littermates had pathologic alterations relevant to hepatotumorigenesis in even up to 52 wk. CONCLUSION: HBV x gene and nodularin promote the development of AFB(1)-induced liver tumors. Co-exposure to AFB1 and MC-LR tends to elevate the risk of liver tumors at 24 wk relative to exposure to one of them. The combinative effect of AFB1, cyanotoxins and HBVx on hepatotumorigenesis is weak at 24 wk. PMID- 16718790 TI - Alterations of seminal and hormonal parameters: An extrahepatic manifestation of HCV infection? AB - AIM: To evaluate the possible influences of HCV infection and relative antiviral treatment on seminal parameters and reproductive hormonal serum levels. METHODS: Ten male patients with HCV-related chronic hepatitis and 16 healthy male volunteers were studied. In all subjects seminal parameters (nemaspermic concentration, progressive motility, morphology) and hormonal levels were determined. Seminal parameters and inhibin B, follicle-stimulating hormone, luteinizing hormone, total and free testosterone, estradiol, prolactine in patients were measured after six and twelve months of antiviral combined (interferon+ribavirin) treatment. RESULTS: Patients before treatment showed a significantly lower nemaspermic motility and morphology as well as lower inhibin B and free testosterone levels than controls. Inhibin B levels in cases were improved six and 12 mo after treatment in five responders (161.9+/-52.8 pg/mL versus 101.7+/-47.0 pg/mL and 143.4+/-46.1 pg/mL versus 95.4+/-55.6 pg/mL, respectively). Hormonal pattern of patients did not significantly change after treatment, with the exception of estradiol levels with an initial reduction and an overall subsequent increment (19.7+/-6.4 pg/mL versus 13.6+/-5.0 pg/mL versus 17.3+/-5.7 pg/mL). However in 1-year responders a significant increment of free testosterone (14.2+/-2.54 pg/mL versus 17.1+/-2.58 pg/mL) occurred. An impairment of nemaspermic morphology occurred, while other seminal parameters did not change significantly during antiviral treatment. CONCLUSION: Patients with HCV infection show worse spermatic parameters than controls, suggesting a possible negative influence of virus on spermatogenesis, with further mild impairment during antiviral treatment. However therapy could improve the spermatic function, as suggested by the increased inhibin B levels and improved hormonal pattern in responders. Further studies are needed to confirm these preliminary intriguing results. PMID- 16718791 TI - Production of reactive oxygen species and expression of inducible nitric oxide synthase in rat isolated Kupffer cells stimulated by Leptospira interrogans and Borrelia burgdorferi. AB - AIM: To evaluate the production of reactive oxygen species (ROS) and the expression of inducible nitric oxide synthase (iNOS) in rat isolated Kupffer cells (KCs) stimulated by Leptospira interrogans and Borrelia burgdorferi. METHODS: Rat Kupffer cells were separated by perfusion of the liver with 0.05% collagenase, and purified by Percoll gradients. Purified Kupffer cells were tested in vitro with alive L. interogans and B. burgdorferi preparations. The production of ROS was determined by chemiluminescence, whereas iNOS protein expression was evaluated by Western blot assay using anti-iNOS antibodies. RESULTS: B. burgdorferi and to a less extent L. interrogans induced ROS production with a peak 35 min after infection. The chemiluminescence signal progressively diminished and was undetectable by 180 min of incubation. Leptospirae and borreliae induced an increased iNOS expression in Kupffer cells that peaked at 6 hours and was still evident 22 h after infection. CONCLUSION: Both genera of spirochetes induced ROS and iNOS production in rat Kupffer cells. Since the cause of liver damage both in leptospiral as well as in borrelial infections are still unknown, we suggest that leptospira and borrelia damage of the liver can be initially mediated by oxygen radicals, and is then maintained at least in part by nitric oxide. PMID- 16718792 TI - Depressed-type (0-IIc) colorectal neoplasm in patients with family history of first-degree relatives with colorectal cancer: A cross-sectional study. AB - AIM: To investigate the correlation of depressed-type (0-IIc) colorectal neoplasm and family history of first-degree relatives (FDR) with colorectal cancer (CRC). METHODS: This cross-sectional study was conducted from June 2000 to October 2002 at National Cancer Center Hospital East. Eligible patients undergoing initial total colonoscopy were surveyed regarding family history of CRC among FDR by a questionnaire prior to colonoscopic examinations. All endoscopic findings during colonoscopy were recorded and the macroscopic classification of the early stage neoplasm/cancer was classified into two types (0-IIc vs non 0-IIc). Odds ratios (OR) and 95% confidence intervals (CI) were calculated by univariate and multivariate logistic regression to estimate the association between macroscopic features and clinicopathological data including gender, age, and family history of FDR with CRC. RESULTS: The OR of an association between family history of FDR with CRC and overall early stage neoplasm adjusted by gender and age was 1.85 (95% CI: 1.31-2.61, P = 0.0004), that for non 0-IIc neoplasm was 1.71 (95% CI: 1.22-2.41, P = 0.0017) and for 0-IIc colorectal neoplasm was 2.78 (95% CI: 1.49 5.16, P = 0.0031). CONCLUSION: Our study shows a significant association between a family history of FDR with CRC and 0-IIc colorectal neoplasm. When patients with a family history of FDR with CRC undergo colonoscopy, colonoscopists should check carefully for not only polypoid, but also depressed-type (0-IIc) lesions. PMID- 16718793 TI - Hydrogen and methane gases are frequently detected in the stomach. AB - AIM: To investigate the incidence of bacterial overgrowth in the stomach by using a new endoscopic method in which intragastric hydrogen and methane gases are collected and analyzed. METHODS: Studies were performed in 490 consecutive patients undergoing esophagogastroscopy. At endoscopy, we intubated the stomach without inflation by air, and 20 mL of intragastric gas was collected through the biopsy channel using a 30 mL syringe. Intragastric hydrogen and methane concentrations were immediately measured by gaschromatography. H pylori infection was also determined by serology. RESULTS: Most of intragastric hydrogen and methane levels were less than 15 ppm (parts per million). The median hydrogen and methane values (interquartile range) were 3 (1-8) ppm and 2 (1-5) ppm, respectively. The high hydrogen and methane levels for indication of fermentation were decided if the patient had the values more than 90 percentile range in each sample. When a patient had a high level of hydrogen or methane in one or more samples, the patient was considered to have fermentation. The overall incidence of intragastric fermentation was 15.4% (73/473). Intragastric methane levels were higher in the postoperative group than in other groups. None of the mean hydrogen or methane values was related to H pylori infection. CONCLUSION: Hydrogen and methane gases are more frequently detected in the stomach than expected, regardless of the presence of abdominal symptoms. Previous gastric surgery influences on the growth of methane-producing bacteria in the fasting stomach. PMID- 16718794 TI - Seventy-five gram glucose tolerance test to assess carbohydrate malabsorption and small bowel bacterial overgrowth. AB - AIM: To investigate non-invasively the incidence of absorption of carbohydrates in diabetic patients during an oral glucose tolerance test (OGTT) and to determine whether malabsorption may be associated with insulin secretion and insulin resistance. METHODS: A standard 75-g OGTT was performed in 82 diabetic patients. The patients received 75 g of anhydrous glucose in 225 mL of water after an overnight fasting and breath samples were collected at baseline and up to 120 min after ingestion. Breath hydrogen and methane concentrations were measured. Blood glucose and serum insulin concentrations were measured before ingestion and at 30, 60, 90, 120 min post-ingestion. RESULTS: When carbohydrate malabsorption was defined as subjects with an increase of at least 10 ppm (parts per million) in hydrogen or methane excretion within a 2-h period, 28 (34%) had carbohydrate malabsorption. According to the result of increased breath test, 21 (75%) patients were classified as small bowel bacterial overgrowth and 7 (25%) as glucose malabsorption. Patients with carbohydrate malabsorption were older and had poor glycemic control as compared with those without carbohydrate malabsorption. The HOMA value, the sum of serum insulin during the test and the Delta insulin/Delta glucose ratio were greater in patients with carbohydrate malabsorption. CONCLUSION: Insulin resistance may be overestimated by using these markers if the patient has carbohydrate malabsorption, or that carbohydrate malabsorption may be present prior to the development of insulin resistance. Hence carbohydrate malabsorption should be taken into account for estimating insulin resistance and beta-cell function. PMID- 16718795 TI - Low serum levels of ghrelin are associated with gallstone disease. AB - AIM: To explore the role of ghrelin in gallstone disease. METHODS: We carried out a cross-sectional study in 150 subjects, 38 with gallstones (cases) and 112 controls. We also did a real-time PCR-RT study in twenty gallbladder samples each. Body mass index (BMI), serum insulin, ghrelin, and serum lipids were measured. Logistic regression analyses (univariate and multivariate) were conducted to estimate the probability of gallstone disease associated with serum ghrelin concentrations. RESULTS: Cases were statistically different from controls in gender distribution (P = 0.01), age (53 vs 44 yr, P = 0.002), BMI (28 vs 25; P = 0.004), and glucose (5.26 vs 4.98 mmol/L; P = 0.05). The prevalence of ghrelin serum levels above the third tercile was lower in subjects without metabolic syndrome (P < 0.05). In a multivariate model, we found a protective effect, when ghrelin values were higher than the median value (OR = 0.27, 95%CI 0.09-0.82, P = 0.02). Twenty (20%) gallbladder specimens expressed ghrelin mRNA. CONCLUSION: Serum ghrelin concentrations are associated with a protective effect of GD. PMID- 16718796 TI - Ileoscopy in 39 hematochezia patients with normal colonoscopy. AB - AIM: To assess the role of retrograde terminal ileoscopy in hematochezia patients with normal colonoscopy. METHODS: Between January 1997 and March 2005, 39 hematochezia patients (males 36, females 3, mean age 44.7 years) with a reported normal colonoscopy underwent a repeat colonoscopy. After reaching the cecum, attempt was made to localize the ileocecal valve and intubate the terminal ileum. Any abnormality in the mucosa of the terminal ileum was carefully recorded and biopsies were obtained from suspicious-looking lesions. RESULTS: During the study period there were 39 patients admitted for hematochezia in whom colonoscopy till cecum did not reveal any abnormality. Full-length colonoscopy till the cecum could be performed in all the patients. The terminal ileum could be intubated in 36 patients. No abnormality was noted in 31 patients. Ileal ulcers were noted in two patients. Nodularity along with ulceration of the ileal mucosa, a Dieulafoy's lesion, and an angiomatous malformation were noted in one patient each. Histological examination of the biopsies obtained from the ulcers revealed typical tuberculous lesion in the patient with nodularity and ulceration. One of the patients had typhoid ulcers and another had non-specific ulcers. CONCLUSION: Retrograde terminal ileoscopy gives limited but valuable information, in patients with hematochezia and should be attempted in all such patients. PMID- 16718797 TI - Efficacy of one-day quadruple therapy for H pylori infection in Chinese patients. AB - AIM: To compare the efficacies of one-day quadruple therapy and seven-day triple therapy in Chinese patients. METHODS: Sixty consecutive patients with nonulcer dyspepsia and confirmed H pylori infection were randomized to receive either omeprazole 40 mg, amoxycillin 1 g, and furazolidone 100 mg, all twice a day for 7 d or omeprazole 20 mg (at breakfast and dinner), amoxicillin 1 g, furazolidone 200 mg, and colloidal bismuth subcitrate 220 mg four times for only one day. H pylori status was determined before and at least 5 weeks after therapy by endoscopy with antral and corpus biopsies for rapid urease test and histology. RESULTS: H pylori eradication was successful in 66.67% (20/30) patients in the 7 d group and in 36.67% (11/30) patients in the 1-d group (P = 0.037). Side effects were induced by the treatment in 13.3% (4/30) patients of each group, but these were all self-limiting, short-lasting, and did not require any specific treatment. CONCLUSION: The one-day quadruple therapy is less effective than the one-week regimen in curing H pylori infection in Chinese patients. PMID- 16718798 TI - Optimal injection volume of epinephrine for endoscopic treatment of peptic ulcer bleeding. AB - AIM: To define the optimal injection volume of epinephrine with high efficacy for hemostasis and low complication rate in patients with actively bleeding ulcers. METHODS: This prospective, randomized, comparative trial was conducted in a medical center. A total of 228 patients with actively bleeding ulcers (spurting or oozing) were randomly assigned to three groups with 20, 30 and 40 mL endoscopic injections of an 1:10000 solution of epinephrine. The hemostatic effects and clinical outcomes were compared between the three groups. RESULTS: There were no significant differences in all background variables between the three groups. Initial hemostasis was achieved in 97.4%, 98.7% and 100% of patients respectively in the 20, 30 and 40 mL epinephrine groups. There were no significant differences in the rate of initial hemostasis between the three groups. The rate of peptic ulcer perforation was significantly higher in the 40 mL epinephrine group than in the 20 and 30 mL epinephrine groups (P < 0.05). The rate of recurrent bleeding was significantly higher in the 20 mL epinephrine group (20.3%) than in the 30 (5.3%) and 40 mL (2.8%) epinephrine groups (P < 0.01). There were no significant differences in the rates of surgical intervention, the amount of transfusion requirements, the days of hospitalization, the deaths from bleeding and 30 d mortality between the three groups. The number of patients who developed epigastric pain due to endoscopic injection, was significantly higher in the 40 mL epinephrine group (51/76) than in the 20 (2/76) and 30 mL (5/76) epinephrine groups (P < 0.001). Significant elevation of systolic blood pressure after endoscopic injection was observed in the 40 mL epinephrine group (P < 0.01). Significant decreasing and normalization of pulse rates after endoscopic injections were observed in the 20 mL and 30 mL epinephrine groups (P < 0.01). CONCLUSION: Injection of 30 mL diluted epinephrine (1:10000) can effectively prevent recurrent bleeding with a low rate of complications. The optimal injection volume of epinephrine for endoscopic treatment of an actively bleeding ulcer (spurting or oozing) is 30 mL. PMID- 16718799 TI - Conversion to sirolimus immunosuppression in liver transplantation recipients with hepatocellular carcinoma: Report of an initial experience. AB - AIM: To report a retrospective analysis of preliminary results of 36 patients who received sirolimus (SRL, Rapamune, rapamycin) in a consecutive cohort of 248 liver allograft recipients. METHODS: Thirty-six liver transplant patients with hepatocellular carcinoma (HCC) who were switched to SRL-based immunosuppression therapy from tacrolimus were enrolled in this study. The patients who were diagnosed as advanced HCC before orthotopic liver transplantation (OLT) were divided into group A (n = 11), those who were found to have HCC recurrence and/or metastasis after OLT were assigned to group B (n = 18), and those who developed renal insufficiency caused by calcineurin inhibitor (CNI) were assigned to group C (n = 7) after OLT. RESULTS: The patients were followed up for a median of 10.4 mo (range, 3.8-19.1 mo) after conversion to SRL therapy and 12.3 mo (range, 5.1 34.4 mo) after OLT. Three patients developed mild acute cellular rejection 2 wk after initiating SRL therapy, which was fully reversed after prednisolone pulse therapy. In group A, only 1 patient was found to have HCC recurrence and metastasis 12 mo after OLT. In group B, 66.7% (12/18) patients (2 with progressive tumor, 7 with stable tumor and 3 without tumor) were still alive due to conversing to SRL and/or resection for HCC recurrence at the end of a median follow-up of 6.8 mo post conversion and 10.7 mo posttransplant. In group C, no HCC recurrence was demonstrated in 7 patients, and renal function became normal after SRL therapy. Thrombocytopenia (n = 2), anemia (n = 8), and oral aphthous ulcers (n = 7) found in our cohort were easily manageable. CONCLUSION: The conversion to SRL-based immunosuppression may inhibit the recurrence and metastasis of HCC and improve CNI-induced renal insufficiency in OLT patients with HCC. PMID- 16718800 TI - Expression of trefoil factors 1 and 2 in precancerous condition and gastric cancer. AB - AIM: To study the expression of trefoil factor 1 (TFF1) and TFF2 in precancerous condition and gastric cancer and to explore the relationship between TFFs and tumorigenesis, precancerous condition and gastric cancer. METHODS: The expression of TFF1 and TFF2 was immunohistochemically analyzed in paraffin-embedded samples from 140 patients including 35 cases of chronic superficial gastritis (CSG), 35 cases of gastric ulcer (GU), 35 cases of chronic atrophic gastritis (CAG) and 35 cases of gastric cancer (GC). RESULTS: TFF1 and TFF2 were located in cytoplasm of gastric mucous cells. In CSG, GU, CAG and GC, the level of TFF1 expression had a decreased tendency (P < 0.05). The expression of TFF2 was higher in GU than in CSG, but the difference was not significant. The expression of TFF2 also had a decreased tendency in GU, CAG, and GC (P < 0.05). CONCLUSION: The reduced expression of TFF1 and TFF2 in precancerous conditions and gastric cancer may be associated with the proliferation and malignant transformation of gastric mucosa. More investigations are needed to explore the mechanism of TFFs and the relationship between TFFs and gastric cancer. PMID- 16718801 TI - Primary shunt hyperbilirubinaemia in a large four-generation family confirming autosomal dominant genetic disorder. AB - AIM: To describe the pattern of inheritance and confirm the diagnostic criteria of primary shunt hyperbilirubinaemia (PSH). METHODS: Forty members of a family pedigree across four generations were included in this study. All family members were interviewed and investigated by physical examination, hematology and liver function test and the pattern of inheritance was analyzed. RESULTS: Nine of the forty family members suffered primary shunt hyperbilirubinaemia. The mature erythrocytes of the propositus were irregular in shape and size. The pedigree showed transmission of the trait through four generations with equal distribution in male and female. No individual with a primary shunt hyperbilirubinaemia was born to unaffected parents. The penetrance was complete in adult. CONCLUSION: The pattern of inheritance is autosomal dominant. The abnormality of erythrocytes and decrease in white blood cell could be supplemented in the diagnosis of PSH. The PSH is a genetic disorder and could by renamed as hereditary shunt hyperbilirubinaemia. PMID- 16718802 TI - This year's Nobel Prize to gastroenterology: Robin Warren and Barry Marshall awarded for their discovery of Helicobacter pylori as pathogen in the gastrointestinal tract. PMID- 16718803 TI - Experimental human pain models in gastro-esophageal reflux disease and unexplained chest pain. AB - Methods related to experimental human pain research aim at activating different nociceptors, evoke pain from different organs and activate specific pathways and mechanisms. The different possibilities for using mechanical, electrical, thermal and chemical methods in visceral pain research are discussed with emphasis of combinations (e.g., the multimodal approach). The methods have been used widely in assessment of pain mechanisms in the esophagus and have contributed to our understanding of the symptoms reported in these patients. Hence abnormal activation and plastic changes of central pain pathways seem to play a major role in the symptoms in some patients with gastro-esophageal reflux disease and in patients with functional chest pain of esophageal origin. These findings may lead to an alternative approach for treatment in patients that does not respond to conventional medical or surgical therapy. PMID- 16718804 TI - Functional oesophago-gastric junction imaging. AB - Despite its role in disease there is still no definitive method to assess oesophago-gastric junction competence (OGJ). Traditionally the OGJ has been assessed using manometry with lower oesophageal sphincter pressure as the indicator. More recently this has been shown not to be a very reliable marker of sphincter function and competence against reflux. Disorders such as gastro oesophageal reflux disease and to a lesser extend achalasia still effects a significant number of patients. This review looks at using a new technique known as impedance planimetry to profile the geometry and pressure in the OGJ during distension of a bag. The data gathered can be reconstructed into a dynamic representation of OGJ action. This has been shown to provide a useful representation of the OGJ and to show changes to the competence of the OGJ in terms of compliance and distensibility as a result of endoluminal therapy. PMID- 16718805 TI - Gastric accommodation assessed by ultrasonography. AB - Gastric accommodation is important for the understanding of the pathophysiology in functional dyspepsia and is also relevant for symptom generation in other disorders. The term gastric accommodation has at least three different meanings: The accommodation process, the accommodation reflex, and the accommodation response. The gastric accommodation process is a complex phenomenon that describes how the size of the gastric compartment changes in response to a meal. The electronic barostat is considered the gold standard in assessing gastric accommodation. Imaging methods, including MRI, SPECT, and ultrasonography may also be used, particularly in patients who are stress-responsive, e.g. functional dyspepsia patients, as a non-invasive and less stress-inducing method is favourable. Ultrasonography satisfies these criteria as it does not by itself distort the physiological response in stress-responsive individuals. PMID- 16718806 TI - Functional findings in irritable bowel syndrome. AB - The pathophysiology of IBS is complex and still incompletely known. Both central and peripheral factors, including psychosocial factors, abnormal GI motility and secretion, and visceral hypersensitivity, are thought to contribute to the symptoms of IBS. Several studies have demonstrated altered GI motor function in IBS patients and the pattern differs between IBS subgroups based on the predominant bowel pattern. Few studies have so far addressed GI secretion in IBS, but there are some evidence supporting altered secretion in the small intestine of IBS patients. Visceral hypersensitivity is currently considered to be perhaps the most important pathophysiological factor in IBS. Importantly, several external and internal factors can modulate visceral sensitivity, as well as GI motility, and enhanced responsiveness within the GI tract to for instance stress and nutrients has been demonstrated in IBS patients. Today IBS is viewed upon as a disorder of dysregulation of the so-called brain-gut axis, involving abnormal function in the enteric, autonomic and/or central nervous systems, with peripheral alterations probably dominating in some patients and disturbed central processing of signals from the periphery in others. PMID- 16718807 TI - Evaluation of the biliary tract in patients with functional biliary symptoms. AB - The aim of this paper was to describe functional biliary syndromes and methods for evaluation of the biliary tract in these patients. Functional biliary symptoms can be defined as biliary symptoms without demonstrable organic substrate. Two main syndromes exist: Gallbladder dysfunction and sphincter of Oddi dysfunction. The most important investigative tools are cholescintigraphy and endoscopic sphincter of Oddi manometry. In gallbladder dysfunction a scintigraphic gallbladder ejection fraction below 35% can select patients who will benefit from cholecystectomy. Endoscopic sphincter of Oddi manometry is considered the gold standard in sphincter of Oddi dysfunction but recent development in scintigraphic methods is about to change this. Thus, calculation of hilum-to-duodenum transit time and duodenal appearance time on cholescintigraphy have proven useful in these patients. In conclusion, ambient methods can diagnose functional biliary syndromes. However, there are still a number of issues where further knowledge is needed. Probably the next step forward will be in the area of sensory testing and impedance planimetric methods. PMID- 16718809 TI - Morphology and motor function of the gastrointestinal tract examined with endosonography. AB - Endosonography is a useful tool for studying the morphology and motor function of the gastrointestinal tract. Intraluminal ultrasonography is the common denomination of ultrasound examinations using intracorporal transducers which are inserted into the GI tract. Thus, the visceral wall and adjacent structures can be imaged in detail. This review describes the usefulness of endosonography in gastroenterology, in particular with respect to studies of the biomechanical and motor function of the gastrointestinal tract. New techniques such as 3-D EUS, elastography and strain rate imaging are discussed. PMID- 16718808 TI - Upper gastrointestinal sensory-motor dysfunction in diabetes mellitus. AB - Gastrointestinal (GI) sensory-motor abnormalities are common in patients with diabetes mellitus and may involve any part of the GI tract. Abnormalities are frequently sub-clinical, and fortunately only rarely do severe and life threatening problems occur. The pathogenesis of abnormal upper GI sensory-motor function in diabetes is incompletely understood and is most likely multi factorial of origin. Diabetic autonomic neuropathy as well as acute suboptimal control of diabetes has been shown to impair GI motor and sensory function. Morphological and biomechanical remodeling of the GI wall develops during the duration of diabetes, and may contribute to motor and sensory dysfunction. In this review sensory and motility disorders of the upper GI tract in diabetes is discussed; and the morphological changes and biomechanical remodeling related to the sensory-motor dysfunction is also addressed. PMID- 16718810 TI - New perspectives of studying gastrointestinal muscle function. AB - The motor function of the gastrointestinal tract has primarily been studied using manometry and radiography, though more indirect tests have also been applied. Manometry and radiography do not provide detailed information about the muscle properties as can be assessed from studies of muscle properties in muscle strips in vitro. In recent years a technique based on impedance planimetric measurement of pressure-cross-sectional area relations in a distending bag has proven to provide more detailed information about the muscle function in vivo. This review shows examples of new muscle function analysis such as length-tension diagrams, force-velocity curves and preload-afterload diagrams. PMID- 16718811 TI - Three-dimensional surface model analysis in the gastrointestinal tract. AB - The biomechanical changes during functional loading and unloading of the human gastrointestinal (GI) tract are not fully understood. GI function is usually studied by introducing probes in the GI lumen. Computer modeling offers a promising alternative approach in this regard, with the additional ability to predict regional stresses and strains in inaccessible locations. The tension and stress distributions in the GI tract are related to distensibility (tension strain relationship) and smooth muscle tone. More knowledge on the tension and stress on the GI tract are needed to improve diagnosis of patients with gastrointestinal disorders. A modeling framework that can be used to integrate the physiological, anatomical and medical knowledge of the GI system has recently been developed. The 3-D anatomical model was constructed from digital images using ultrasonography, computer tomography (CT) or magnetic resonance imaging (MRI). Different mathematical algorithms were developed for surface analysis based on thin-walled structure and the finite element method was applied for the mucosa-folded three layered esophageal model analysis. The tools may be useful for studying the geometry and biomechanical properties of these organs in health and disease. These studies will serve to test the structure-function hypothesis of geometrically complex organs. PMID- 16718812 TI - Immune therapy including dendritic cell based therapy in chronic hepatitis B virus infection. AB - Hepatitis B virus (HBV) infection is a global public health problem. Of the approximately 2 billion people who have been infected worldwide, more than 400 million are chronic carriers of HBV. Considerable numbers of chronic HBV carriers suffer from progressive liver diseases. In addition, all HBV carriers are permanent source of this virus. There is no curative therapy for chronic HBV carriers. Antiviral drugs are recommended for about 10% patients, however, these drugs are costly, have limited efficacy, and possess considerable side effects. Recent studies have shown that immune responses of the host to the HBV are critically involved at every stage of chronic HBV infection: (1) These influence acquisition of chronic HBV carrier state, (2) They are important in the context of liver damages, (3) Recovery from chronic HBV-related liver diseases is dependent on nature and extent of HBV-specific immune responses. However, induction of adequate levels of HBV-specific immune responses in chronic HBV carriers is difficult. During the last one decade, hepatitis B vaccine has been administered to chronic HBV carriers as a therapeutic approach (vaccine therapy). The present regimen of vaccine therapy is safe and cheap, but not so effective. A dendritic cell-based therapeutic vaccine has recently been developed for treating chronic HBV infection. In this review, we will discuss about the concept, scientific logics, strategies and techniques of development of HBV-specific immune therapies including vaccine therapy and dendritic cell-based vaccine therapy for treating chronic HBV infection. PMID- 16718813 TI - Non-steroidal anti-inflammatory drugs in prevention of gastric cancer. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) including cyclooxygenase 2 (COX-2) selective inhibitors, are potential agents for the chemoprevention of gastric cancer. Epidemiological and experimental studies have shown that NSAID use is associated with a reduced risk of gastric cancer although many questions remain unanswered such as the optimal dose and duration of treatment. The possible mechanisms for the suppressor effect of NSAIDs on carcinogenesis are the ability to induce apoptosis in epithelial cells and regulation of angiogenesis. Both COX dependent and COX-independent pathways have a role in the biological activity of NSAIDs. Knowledge of how NSAIDs prevent neoplastic growth will greatly aid the design of better chemopreventive drugs and novel treatments for gastric cancer. PMID- 16718814 TI - Anticancer effect of jinlongshe granules on in situ-transplanted human MKN-45 gastric cancer in nude mice and xenografted sarcoma 180 in Kunming mice and its mechanism. AB - AIM: To study the antitumor effect of Chinese compound jinlongshe (JLS) granules on sarcoma 180 and MKN-45 human gastric cancer cell lines in vivo and its mechanism. METHODS: After establishment of S180 sarcoma (S180) and MKN-45 gastric cancer model of nude mice, the tumor-bearing mice were divided into 5 groups at random. Three experimental groups were respectively given the aqueous extract of JLS granules at doses of 120 g, 60 g and 20 g/(kg per 6/wk, i.g.) for 3 wk in S180 and 6 wk in nude mice model. Positive control was given cyclophosphamide (Cy) at a dose of 50 mg/(kg per 3/wk, i.g.) for 3 wk in S180 models and 5 Fluorouracil (5-FU) 20 mg/(kg per 3/wk, i.g.) for 3 wk in nude mice model. Negative control was given normal saline (NS) at a dose of 0.18 g/(kg per 6/wk, i.g.) respectively. After 3 wk in mice bearing S180 tumor and 6 wk in nude mice model, the experimental animals were sacrificed and the masses of tumor were weighed, and the rates of tumor inhibition of each treated group were calculated respectively. To determine the antitumor mechanisms, the morphological changes, cell cycle and apoptosis were observed in MKN-45 nude mice model. Annexin V FITC/PI double staining FCM assay was used to further determine the live cells, apoptotic cells, necrotic cells and debris. RESULTS: The inhibitory rates of JLS granules at the doses of 20 g/kg, 60 g/kg and 120 g/kg were 50.31%, 55.94% and 68.13% (P < 0.01) in nude mice models and 40.90%, 50.32% and 58.46% (P < 0.01) in S180 model. The inhibitory rate of Cy was 85.22% in S180 models and the inhibitory rate of 5-FU was 53.43% in nude mice model (P < 0.01). Nuclear chromatin and margination were observed under a transmission electron microscope (TEM). The G0/G1 phase was arrested, typical apoptotic peak appeared, the apoptotic rate was 22.81%-38.54% in three JLS granule-treated groups. Annexin V FITC/PI double staining FCM assay showed that the apoptotic cells were 4.36%, 3.08% and 7.08% in three dosages, most cells were localized in the low right quadrant. CONCLUSION: Jinlongshe granules possess anti-tumor effects on experimental tumor models in vivo, and apoptosis induction is one of its anti tumor mechanisms. PMID- 16718815 TI - Partial Beclin 1 silencing aggravates doxorubicin- and Fas-induced apoptosis in HepG2 cells. AB - AIM: To investigate the role of Beclin 1 on the susceptibility of HepG2 cells to undergo apoptosis after anti-Fas antibody or doxorubicin treatment. METHODS: Beclin 1 silencing was achieved using RNA interference. DNA ploidy, the percentage of apoptotic cells and the mitochondrial membrane potential were assessed by flow cytometry. Levels of Beclin 1, Bcl-X(L) and cytochrome c, and the cleavage of poly (ADP-ribose) polymerase (PARP) were assayed by using Western blots. RESULTS: Beclin 1 expression decreased by 75% 72 h after Beclin 1 siRNA transfection. Partial Beclin 1 silencing significantly increased the percentage of subG1 cells 24 and 40 h after treatment with doxorubicin or anti-Fas antibody, respectively, and this potentiation was abrogated by treatment with a pan-caspase inhibitor. Partial Beclin 1 silencing also increased PARP cleavage, mitochondrial membrane depolarization and cytosolic cytochrome c. The pro-apoptotic consequences of partial Beclin 1 silencing were not associated with a decline in Bcl-X(L) expression. CONCLUSION: Partial Beclin 1 silencing aggravates mitochondrial permeabilization and apoptosis in HepG2 cells treated with an anti Fas antibody or with doxorubicin. PMID- 16718816 TI - Down-regulation of survivin expression by small interfering RNA induces pancreatic cancer cell apoptosis and enhances its radiosensitivity. AB - AIM: To investigate the inhibitory effect of small interfering RNA (siRNA) on the expression of survivin in pancreatic cancer cell line PC-2 and the role of siRNA in inducing PC-2 cell apoptosis and enhancing its radiosensitivity. METHODS: A siRNA plasmid expression vector against survivin was constructed and transfected into PC-2 cells with Lipofectamine 2000. The down regulation of survivin expression was detected by semi-quantitative RT-PCR and immunohistochemical SP method and the role of siRNA in inducing PC-2 cell apoptosis and enhancing its radiosensitivity was detected by flow cytometry. RESULTS: The sequence-specific siRNA efficiently and specifically down-regulated the expression of survivin at both mRNA and protein levels. The expression inhibition ratio was 81.25% at mRNA level detected by semi-quantitative RT-PCR and 74.24% at protein level detected by immunohistochemical method. Forty-eight hours after transfection,apoptosis was induced in 7.03% cells by siRNA and in 14.58% cells by siRNA combined with radiation. CONCLUSION: The siRNA plasmid expression vector against survivin can inhibit the expression of survivin in PC-2 cells efficiently and specifically. Inhibiting the expression of survivin can induce apoptosis of PC-2 cells and enhance its radiosensitivity significantly. RNAi against survivin is of potential value in gene therapy of pancreatic cancer. PMID- 16718817 TI - Hyperlipidemia intensifies cerulein-induced acute pancreatitis associated with activation of protein kinase C in rats. AB - AIM: To investigate the effects of hyperlipidemia on acute pancreatitis (AP) and the possible mechanisms. METHODS: Rat models of hyperlipidemia and AP were established by Triton WR1339 and cerulein respectively. Human albumin was used to treat AP complicated by hyperlipidemia. In each group, we compared the histological score, volume of ascites, ratio of pancreatic wet/dry weight, serum amylase (AMY) and pancreatic acinar cell apoptosis. The level of protein kinase C (PKC) membrane translocation in pancreatic tissue was detected by Western blot. RESULTS: In the hyperlipidemia model established by Triton WR1339, triglyceride (TG) increased remarkably and reached its peak 6 h after injection, and most rats developed mild acute pancreatitis. Histological score, volume of ascites, ratio of wet/dry weight and serum AMY in AP animals with hyperlipidemia were obviously higher than those in AP animals (P < 0.05) and decreased after albumin therapy but not significantly (P > 0.05). Apoptotic cells detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) increased in AP animals with hyperlipidemia and did not change distinctly after albumin therapy. PKC membrane translocation level increased in AP animals with hyperlipidemia and decreased remarkably after albumin therapy (P < 0.05). CONCLUSION: Hyperlipidemia may induce AP or intensify pancreatic injury. Albumin therapy can not alleviate pancreatic lesion effectively. PKC activation may be one mechanism by which AP is intensified by hyperlipidemia. PMID- 16718818 TI - Oxygen radical formation does not have an impact in the treatment of severe acute experimental pancreatitis using free cellular hemoglobin. AB - AIM: Microcirculatory dysfunction and free oxygen radicals are important factors in the pathogenesis of severe acute pancreatitis. Additional oxygen delivery might enhance lipid peroxidation but may also improve pancreatic microcirculation. This study assesses the effect of free cellular bovine hemoglobin on the formation of oxygen radicals and microcirculation in a rodent model of severe acute pancreatitis. METHODS: Fifteen minutes after induction of acute pancreatitis Wistar rats received either 0.8 mL bovine hemoglobin (HBOC 200), hydroxyethyl starch (HES) or 2.4 mL of normal saline to ensure normovolemic substitution. After 6 h of examination the pancreas was excised and rapidly processed for indirect measurement of lipid peroxidation products malondialdehyde (MDA) and reduced glutathione (GSH) in pancreatic tissue. RESULTS: The single application of HBOC-200 improved pancreatic microcirculation and reduced histopathological tissue damage significantly. Tissue concentration of MDA did not differ between the groups. Also no differences in GSH levels were detected. CONCLUSION: Though the single application of HBOC-200 and HES improve pancreatic microcirculation, no differences in lipid peroxidation products were detected. The beneficial effect of additional oxygen supply (HBOC-200) does not lead to enhanced lipid peroxidation. PMID- 16718819 TI - MRCP and MRI findings in 9 patients with autoimmune pancreatitis. AB - AIM: To evaluate magnetic resonance cholangiopancreatography (MRCP) findings in conjunction with magnetic resonance (MR) images in autoimmune pancreatitis (AIP) patients. METHODS: Nine patients with AIP underwent MRI, MRCP, endoscopic retrograde cholangiopancreatography (ERCP), computed tomography, and ultrasonography. The MRCP and MR images taken before and after steroid therapy were reviewed and compared with other imaging modalities. The MRCP findings of the AIP cases were compared to those of 10 cases with carcinoma of the head of the pancreas. RESULTS: On MRCP, the narrowed portion of the main pancreatic duct noted on ERCP was not visualized, while the non-involved segments of the main pancreatic duct were visualized. The degree of upstream dilatation of the proximal main pancreatic duct was milder than that seen in cases of pancreatic carcinoma. Stenosis or obstruction of the lower bile duct was detected in 8 patients. MR images showed enlargement of the pancreas with decreased signal intensity on T1-weighted MR images, increased signal intensity on T2-weighted MR images, and, in 3 patients, a hypointense capsule-like rim. After steroid therapy, the previously not visualized portion of the main pancreatic duct was seen, along with improvement of the bile duct stenosis. Pancreatic enlargement decreased, and the abnormal signal intensity on both T1- and T2-weighted MR images became isointense. CONCLUSION: MRCP cannot differentiate irregular narrowing of the main pancreatic duct seen with AIP from stenosis of the main pancreatic duct seen with pancreatic carcinoma. However, MRCP findings in conjunction with MR imaging of pancreatic enlargement that shows abnormal signal intensity on T1- and T2-weighted MR images are useful in supporting a diagnosis of AIP. PMID- 16718820 TI - Incidence and predictors of upper gastrointestinal bleeding in patients receiving low-dose aspirin for secondary prevention of cardiovascular events in patients with coronary artery disease. AB - AIM: The use of low-dose aspirin to prevent cardiovascular disease events is well established. However, the incidence and predictors of upper gastrointestinal bleeding (UGIB) with its use are unknown. We studied prospectively the incidence and outcome of peptic ulceration in low-dose aspirin users. METHODS: A total of 991 patients with coronary artery disease (CAD) on low-dose aspirin were prospectively followed-up for two years for the occurrence and clinical features of first hospitalized episode of UGIB. RESULTS: UGIB had a bimodal presentation with 45% occurring within four months of aspirin initiation and had an overall prevalence of 1.5% per year. There was no UGIB-related death. Hypertension (OR = 4.6, 95%CI 1.5-14.7, P = 0.009), history of peptic ulceration (OR = 3.1, 95%CI 1.1-9.0, P = 0.039), tertiary education (OR = 3.08, 95%CI 1.1-9.0, P = 0.039) and higher lean body mass (P = 0.016) were independent factors associated with UGIB. Use of nitrate did not reduce UGIB. CONCLUSION: The incidence of UGIB in patients with CAD on long-term low-dose aspirin is low, but is accompanied with significant morbidity. With prolonged use of aspirin, UGIB continues to be a problem for those with risk factors and especially in patients with a history of peptic ulcers, in which UGIB tends to occur early after aspirin therapy. PMID- 16718821 TI - Relationship between IFN-gamma gene polymorphism and susceptibility to intrauterine HBV infection. AB - AIM: To explore the susceptibility of children to intrauterine HBV infection by studying the relationship between IFN-gamma gene polymorphism, including IFN gamma+874A/T single nucleotide polymorphism (SNP) and CA repeat microsatellite polymorphism and intrauterine HBV infection. METHODS: A TaqMan fluorescence polymerase chain reaction in the IFN-gamma+874A/T single nucleotide polymorphism was tested in the intrauterine HBV infection group (group I) and the normal immune children group (group II). Capillary electrophoresis was performed in the above two groups to assay the IFN-gamma CA repeat microsatellite polymorphism. RESULTS: Frequencies of AA, AT and TT genotypes were 67.4%, 19.6% and 13.0% in the intrauterine HBV infection group, and 45.2%, 30.1% and 24.7% in the normal immune children group, respectively. A significant difference was found in the frequency distribution of IFN-gamma+874 genotype between the two groups (chi2 = 5.102, P = 0.02389). In the intrauterine HBV infection group the AA genotype was more common than in the normal immune group. Frequency of IFN-gamma+874A allele was 77.17% in the intrauterine HBV infection group, and 60.27% in the normal immune children group. In the intrauterine HBV infection group the IFN-gamma+874A allele was more common than in normal immune group. A significant difference was found in the frequency distribution between the two groups (chi2 = 7.238, P = 0.02389, OR = 2.228, 95% CI = 1.244-3.992). (CA12)+/(CA12)+ of IFN-gamma CA microsatellite polymorphism was 11.90% in the intrauterine HBV infection group and 26.47% in the normal immune children group. A significant difference was found in the frequency distribution between the two groups (chi2 = 5.64, P = 0.0176). Frequency of IFN-gamma CA repeat was 25% in the intrauterine HBV infection group and 43.38% in the normal immune children group. The frequency of IFN-gamma CA repeat was less in the intrauterine HBV infection group than in normal immune group. A significant difference was found in the frequency distribution between the two groups (chi2 = 7.548, P = 0.0060). CONCLUSION: There is a relationship between IFN-gamma+874A/T SNP and intrauterine HBV infection as well as between IFN-gamma CA microsatellite polymorphism and intrauterine HBV infection. IFN-gamma gene polymorphism might be important in determining individual's susceptibility to intrauterine HBV infection. PMID- 16718822 TI - Diverticulitis in the young patient--is it different ? AB - AIM: To prospectively study the incidence and the natural history of acute diverticulitis in young patients. METHODS: A total of 207 patients hospitalized at our hospital between January 2000 to February 2005 with the diagnosis of acute diverticulitis were included. Their demographic characteristics, medical history, physical, radiographic and endoscopic findings as well as therapy were recorded. Patients were followed every 6 mo for the first year and later annually. RESULTS: The mean patients' age was 61 (range 27-92) years. Twenty-five patients (12%) were younger than 45 years. Acute diverticulitis was significantly more prevalent among male in the young age group as compared to the older age group (19/25, 76% vs 61/182, 33%, respectively, P = 0.0001). Complications occurred more often in the young age group; 32% vs 13%, (P = 0.002). During follow-up, 6 patients (28%) remained asymptomatic in the young age group as compared to 87 patients (55%) in the older age group (P = 0.024). As a result, sigmoidectomies were performed twice as often in the young age group (38% vs 13%, P = 0.002). CONCLUSION: Diverticulitis in young patients has a male predominance, a more aggressive course with a higher rate of complications and a higher recurrence rate. An earlier surgical approach might be considered in young patients with acute diverticulitis. PMID- 16718823 TI - Effects of ischemic preconditioning on cyclinD1 expression during early ischemic reperfusion in rats. AB - AIM: To observe the effect of ischemic preconditioning on cyclinD1 expression in rat liver cells during early ischemic reperfusion. METHODS: Fifty-four SD rats were randomly divided into ischemic preconditioning group (IP), ischemia/reperfusion group (IR) and sham operation group (SO). The IP and IR groups were further divided into four sub-groups (n = 6). Sham operation group (SO) served as the control group (n = 6). A model of partial liver ischemia/reperfusion was used, in which rats were subjected to liver ischemia for 60 min prior to reperfusion. The animals in the IP group underwent ischemic preconditioning twice for 5 min each time prior to the ischemia/reperfusion challenge. After 0, 1, 2, and 4 h of reperfusion, serum and liver tissue in each group were collected to detect the level of serum ALT, liver histopathology and expression of cyclinD1 mRNA and protein. Flow cytometry was used to detect cell cycle as the quantity indicator of cell regeneration. RESULTS: Compared with IR group, IP group showed a significantly lower ALT level in 1 h to 4 h sub-groups (P < 0.05). Proliferation index (PI) indicated by the S-phase and G2/M-phase ratio [(S+G2/M)/(G0/G1+S+G2/M)] was significantly increased in IP group at 0 and 1 h (26.44 +/- 7.60% vs 18.56 +/- 6.40%, 41.87 +/- 7.27% vs 20.25 +/- 6.70%, P < 0.05). Meanwhile, cyclinD1 protein expression could be detected in IP group. But in IR group, cyclinD1 protein expression occurred 2 h after reperfusion. The expression of cyclinD1 mRNA increased significantly in IP group at 0 and 1 h (0.568 +/- 0.112 vs 0.274 +/- 0.069, 0.762 +/- 0.164 vs 0.348 +/- 0.093, P < 0.05). CONCLUSION: Ischemic preconditioning can protect liver cells against ischemia/reperfusion injury, which may be related to cell proliferation and expression of cyclinD1 during early ischemic reperfusion. PMID- 16718824 TI - Lymph node micrometastasis and its correlation with MMP-2 expression in gastric carcinoma. AB - AIM: To examine matrix metalloproteinase-2 (MMP-2) expression in gastric cancer tissues and to evaluate its relationship with lymph node micrometastasis. METHODS: The authors studied 850 lymph nodes resected from 30 patients with gastric carcinoma who underwent gastrectomy with lymphadenectomy using reverse transcription polymerase chain reaction (RT-PCR) assay in addition to H-E staining. MMP-2 expression of the tumor tissues was detected by immunohistochemical technique (EliVision plus). RESULTS: MMP-2 expression was positive in 21 (70%) cases and negative in 9 (30%) cases. No significant correlations were found between MMP-2 expression and other variables such as age, gender, tumor location, tumor diameter, Lauren classification and lymphatic invasion. In contrast, MMP-2 expression correlated significantly with depth of tumor infiltration (P = 0.022), lymph node metastasis (P = 0.030) and tumor differentiation (P = 0.043). Lymph node micrometastases were detected in 77 (12.5%) lymph nodes of 14 (46.7%) gastric carcinoma patients. MMP-2 expression was positive in 12 (85.7%) of the 14 patients with lymph node micrometastasis, and in 9 (56.3%) of the 16 patients without lymph node micrometastasis (P = 0.118). CONCLUSION: Our results demonstrate that MMP-2 expression has significant correlation with tumor invasion, tumor differentiation and lymph node metastases. MMP-2 expression may be an important biological characteristics and significant prognostic parameter of gastric carcinoma. We also conclude that MMP-2 may participate in the development of lymph node micrometastasis of gastric carcinoma. Further investigations are needed to draw a conclusion. PMID- 16718825 TI - Clinical value of CT three-dimensional imaging in diagnosing gastrointestinal tract diseases. AB - AIM: To discuss the clinical value of CT three-dimensional (3-D) imaging in diagnosing gastrointestinal tract diseases. METHODS: Three-D imaging findings of 52 patients were retrospectively analyzed. Three-D imaging methods included shaded surface display (SSD), volume rendering (VR), virtual endoscopy (VE) and multiplanar reformatting (MPR). The diagnosis results of CT 3-D were evaluated by comparison with those of endoscopy and/or surgical finding. RESULTS: Fifty-two patients with gastrointestinal tract diseases were diagnosed by CT 3-D imaging, of whom 50 cases were correctly diagnosed and 2 were misdiagnosed. There were 33 cases of gastric diseases (27 with carcinoma, 5 with peptic ulcer and 1 with leiomyoma) and 19 large intestinal diseases (10 with colon carcinoma, 2 with carcinoma of the rectum, 5 with colon polypus and 2 with tuberculosis of the ileocecal junction). Twenty-two cases with prominent lesions (9 with subsequent hollow lesions), 20 with stenosis of cavity (8 with concomitant prominent lesions) and 10 with hollow lesions (5 with concomitant prominent lesions) were shown in 3-D images. The minimal lesion shown was 1.0 cm x 0.8 cm x 0.5 cm. CONCLUSION: CT 3-D imaging, a non-invasive examination without pain, can display clearly and directly the lesions of gastrointestinal tract with accurate location and high diagnosis accuracy. It is an important complementary technique to endoscopy. PMID- 16718826 TI - Successful aspiration and ethanol sclerosis of a large, symptomatic, simple liver cyst: case presentation and review of the literature. AB - Simple liver cysts are congenital with a prevalence of 2.5%-4.25%. Imaging, whether by US, CT or MRI, is accurate in distinguishing simple cysts from other etiologies, including parasitic, neoplastic, duct-related, and traumatic cysts. Symptomatic simple liver cysts are rare, and the true frequency of symptoms is not known. Symptomatic simple liver cysts are predominantly large (> 4 cm), right sided, and more common in women and older patients. The vast majority of simple hepatic cysts require no treatment or follow-up, though large cysts (> 4 cm) may be followed initially with serial imaging to ensure stability. Attribution of symptoms to a large simple cyst should be undertaken with caution, after alternative diagnoses have been excluded. Aspiration may be performed to test whether symptoms are due to the cyst; however, cyst recurrence should be expected. Limited experience with both laparoscopic deroofing and aspiration, followed by instillation of a sclerosing agent has demonstrated promising results for the treatment of symptomatic cysts. Here, we describe a patient with a large, symptomatic, simple liver cyst who experienced complete resolution of symptoms following cyst drainage and alcohol ablation, and we present a comprehensive review of the literature. PMID- 16718827 TI - Autoimmune pancreatitis metachronously associated with retroperitoneal fibrosis with IgG4-positive plasma cell infiltration. AB - Retroperitoneal fibrosis is an uncommon disorder characterized by the formation of a dense plaque of fibrous tissue in the retroperitoneum, and its etiology remains unknown. Autoimmune pancreatitis is a rare type of chronic pancreatitis characterized by fibrosis with abundant infiltration of IgG4-positive plasma cells and lymphocytes and obliterative phlebitis in the pancreas. We present a case of autoimmune pancreatitis that developed 10 mo after the occurrence of retroperitoneal fibrosis. Histological findings of the resected retroperitoneal mass were marked periureteral fibrosis with abundant infiltration of IgG4 positive plasma cells and lymphocytes and obliterative phlebitis. These findings suggest a common pathophysiological mechanism for retroperitoneal fibrosis and autoimmune pancreatitis in this case. Some cases of retroperitoneal fibrosis might be a retroperitoneal lesion of IgG4-related sclerosing disease. PMID- 16718828 TI - Gastric signet-ring adenocarcinoma presenting with breast metastasis. AB - Breast metastases from gastric cancer are extremely rare. A case report of a 37 year-old female with breast inflammatory invasion and ascites is described. Breast biopsy revealed carcinomatous invasion of the lymphatics from adenocarcinoma cells with signet-ring features. Estrogen (ER) and progesterone receptors (PR) and c-erb-B2 were negative. Upper gastrointestinal endoscopy revealed a prepyloric ulcerative mass. Histopathologic examination of the lesion showed infiltration from a high-grade adenocarcinoma, identical with that of the breast. Immunostaining was positive for cytokeratins CK-7 and CK-20 and CEA and negative for ER and PR. Ascitic fluid cytology was positive for adenocarcinoma cells. Mammography was not diagnostic. Abdominal CT scanning revealed large ovarian masses suggestive of metastases (Krukenberg's tumor). A cisplatin-based regimen was given but no objective response was observed. The patient died six months after initial diagnosis. A review of the literature is performed. PMID- 16718829 TI - Multiple pyogenic liver abscess. AB - Multiple pyogenic liver abscesses have been rarely described. We report a fatal case of multiple pyogenic liver abscesses affecting a 38-year-old woman requiring surgical drainage. Evolution was marked by occurrence of a septic shock with multi-organ system failure. The patient died 48 h after surgery. Causes, therapeutics and outcome of the disease are discussed. PMID- 16718830 TI - Management of parastomal ulcers. AB - Management of surgically placed ostomies is an important aspect of any general surgical or colon and rectal surgery practice. Complications with surgically placed ostomies are common and their causes are multifactorial. Parastomal ulceration, although rare, is a particularly difficult management problem. We conducted a literature search using MD Consult, Science Direct, OVID, Medline, and Cochrane Databases to review the causes and management options of parastomal ulceration. Both the etiology and treatments are varied. Different physicians and ostomy specialists have used a large array of methods to manage parastomal ulcers; these including local wound care; steroid creams; systemic steroids; and, when conservative measures fail, surgery. Most patients with parastomal ulcers who do not have associated IBD or peristomal pyoderma gangrenosum (PPG) often respond quickly to local wound care and conservative management. Patients with PPG, IBD, or other systemic causes of their ulceration need both systemic and local care and are more likely to need long term treatment and possibly surgical revision of the ostomy. The treatment is complicated, but improved with the help of ostomy specialists. PMID- 16718831 TI - Advances in alimentary tract imaging. AB - Advances in imaging techniques are changing the way radiologists undertake imaging of the gastrointestinal tract and their ability to answer questions posed by surgeons. In this paper we discuss the technological improvements of imaging studies that have occurred in the last few years and how these help to better diagnosing alimentary tract disease. PMID- 16718832 TI - Controversies in the treatment of common anal problems. AB - Treating common benign anal diseases has evolved towards more outpatient procedures with better outcome. However, minimizing post-procedure morbidities such as pain and the avoidance incontinence remain the most significant concerns. We introduce some controversies and highlight the developments in current surgical practice for the treatment of common anal problems. PMID- 16718833 TI - Controversies in the treatment of gastroesophageal reflux and achalasia. AB - The immense success of laparoscopic surgery as an effective treatment of gastroesophageal reflux disease (GERD) and achalasia has established minimal invasive surgery as the gold standard for these two conditions with lower morbidity and mortality, shorter hospital stay, faster convalescence, and less postoperative pain. One controversy in the treatment of GERD evolves around laparoscopic antireflux surgery (LARS) as the preferred treatment for Barrett's esophagus and the procedure's potential to reduce the risk of adenocarcinoma of the esophagus. GERD has also been associated with respiratory symptoms, asthma and laryngeal injury, and a second controversy prompts discussions about whether total or partial fundoplication is the more appropriate treatment for GERD. A new and promising alternative in the treatment of GERD is endoluminal therapy. Three types of this new treatment option will be discussed: radiofrequency energy delivered to the lower esophageal sphincter, the creation of a mechanical barrier at the gastroesophageal junction, and the direct endoscopic tightening of the lower esophageal sphincter. Laparoscopic surgery is discussed not only as a very effective treatment for GERD but also as permanent cure for achalasia. This review analyzes the three most important treatment options for achalasia: medications, pneumatic dilatation, and surgical therapy. Medications as the only true non-invasive option in the treatment of achalasia are not as effective as LARS because of their short half-life and variable absorption due to the poor esophageal emptying. The second treatment option, pneumatic dilatation, involves the stretching of the lower esophagus and is still considered the most effective non-surgical treatment for achalasia. Finally, surgical therapy for achalasia and the two major controversies concerning this laparoscopic treatment are discussed. The first involves the extent to which the myotomy is extended onto the stomach, and the second concerns the necessity and type of antireflux procedure to prevent GERD after myotomy. LARS and laparoscopic Heller myotomy are the agreed upon as the gold standards for surgical treatment of GERD and achalasia, respectively. In the hands of an experienced laparoscopic surgeon both are safe and effective treatments for patients with excellent subjective and objective long-term results with at least 90% patient satisfaction. PMID- 16718834 TI - Choledocholithiasis: evolving standards for diagnosis and management. AB - Cholelithiasis, one of the most common medical conditions leading to surgical intervention, affects approximately 10 % of the adult population in the United States. Choledocholithiasis develops in about 10%-20% of patients with gallbladder stones and the literature suggests that at least 3%-10% of patients undergoing cholecystectomy will have common bile duct (CBD) stones. CBD stones may be discovered preoperatively, intraoperatively or postoperatively Multiple modalities are available for assessing patients for choledocholithiasis including laboratory tests, ultrasound, computed tomography scans (CT), and magnetic resonance cholangiopancreatography (MRCP). Intraoperative cholangiography during cholecystectomy can be used routinely or selectively to diagnose CBD stones. The most common intervention for CBD stones is ERCP. Other commonly used interventions include intraoperative bile duct exploration, either laparoscopic or open. Percutaneous, transhepatic stone removal other novel techniques of biliary clearance have been devised. The availability of equipment and skilled practitioners who are facile with these techniques varies among institutions. The timing of the intervention is often dictated by the clinical situation. PMID- 16718835 TI - Treatment strategies in obstructed defecation and fecal incontinence. AB - Obstructed defecation (OD) and fecal incontinence (FI) are challenging clinical problems, which are commonly encountered in the practice of colorectal surgeons and gastroenterologists. These disorders socially and psychologically distress patients and greatly impair their quality of life. The underlying anatomical and pathophysiological changes are complex, often incompletely understood and cannot always be determined. As a consequence, many medical, surgical, and behavioral approaches have been described, with no panacea. Over the past decade, advances in an understanding of these disorders together with rational and similar methods of evaluation in anorectal physiology laboratories (ARP), radiology studies, and new surgical techniques have led to promising results. In this brief review, we discuss treatment strategies and recent updates on clinical and therapeutic aspects of obstructed defecation and fecal incontinence. PMID- 16718836 TI - Gastrointestinal manifestations of endocrine disease. AB - The hormonal interactions among the systems throughout the body are not fully understood; many vague clinical symptoms may in fact be manifestations of underlying endocrine diseases. The aim of the following review is to discuss gastrointestinal manifestations of surgically correctable endocrine diseases, focusing on abnormalities of thyroid function, cancer and finally autoimmune diseases. We also review manifestations of pancreatic endocrine tumors, and multiple endocrine neoplasia. PMID- 16718837 TI - Less common neoplasms of the pancreas. AB - Recently, there has been an increased recognition of neoplasms of the pancreas other than ductal adenocarcinoma. Although not as well studied or characterized as pancreatic adenocarcinoma there are many distinct lesions which exhibit diverse biological behaviors and varying degrees of malignancy. These lesions include: endocrine neoplasms, cystic tumors, solid pseudopapillary tumors, acinar cell carcinoma, squamous cell carcinoma, primary lymphoma of the pancreas, and metastatic lesions to the pancreas. These less common neoplasms are being diagnosed more frequently as the number and sensitivity of diagnostic imaging studies increase. This review article discusses the clinical course, diagnosis, and treatment of these less common, but quite relevant, neoplasms of the pancreas. PMID- 16718838 TI - Modern management of rectal cancer: a 2006 update. AB - The goal of this review is to outline some of the important surgical issues surrounding the management of patients with early (T1/T2 and N0), as well as locally advanced (T3/T4 and/or N1) rectal cancer. Surgery for rectal cancer continues to develop towards the ultimate goals of improved local control and overall survival, maintaining quality of life, and preserving sphincter, genitourinary, and sexual function. Information concerning the depth of tumor penetration through the rectal wall, lymph node involvement, and presence of distant metastatic disease is of crucial importance when planning a curative rectal cancer resection. Preoperative staging is used to determine the indication for neoadjuvant therapy as well as the indication for local excision versus radical cancer resection. Local excision is likely to be curative in most patients with a primary tumor which is limited to the submucosa (T1N0M0), without high-risk features and in the absence of metastatic disease. In appropriate patients, minimally invasive procedures, such as local excision, TEM, and laparoscopic resection allow for improved patient comfort, shorter hospital stays, and earlier return to preoperative activity level. Once the tumor invades the muscularis propria (T2), radical rectal resection in acceptable operative candidates is recommended. In patients with transmural and/or node positive disease (T3/T4 and/or N1) with no distant metastases, preoperative chemoradiation followed by radical resection according to the principles of TME has become widely accepted. During the planning and conduct of a radical operation for a locally advanced rectal cancer, a number of surgical management issues are considered, including: (1) total mesorectal excision (TME); (2) autonomic nerve preservation (ANP); (3) circumferential resection margin (CRM); (4) distal resection margin; (5) sphincter preservation and options for restoration of bowel continuity; (6) laparoscopic approaches; and (7) postoperative quality of life. PMID- 16718839 TI - Update on obesity surgery. AB - The prevalence of obesity in the United States has reached epidemic proportions. With more than 30 million Americans clinically obese, the younger population has also been affected. Surgical therapy should be offered to the severely obese patient who is refractory to nonsurgical therapy, as established by the 1991 NIH Consensus Conference on Gastrointestinal Surgery for Severe Obesity. Surgery is currently the most effective therapy for weight loss. It is far more effective than any other treatment modality, both in terms of the amount of weight loss and in terms of durability in maintaining weight loss. PMID- 16718840 TI - Pediatric inflammatory bowel disease. AB - Inflammatory bowel disease is an important cause of gastrointestinal pathology in children and adolescents. The incidence of pediatric inflammatory bowel disease is increasing; therefore, it is important for the clinician to be aware of the presentation of this disease in the pediatric population. Laboratory tests, radiology studies, and endoscopic procedures are helpful in diagnosing inflammatory bowel disease and differentiating between Crohn's disease and ulcerative colitis. Once diagnosed, the goal of medical management is to induce remission of disease while minimizing the side effects of the medication. Specific attention needs to be paid to achieving normal growth in this susceptible population. Surgical management is usually indicated for failure of medical management, complication, or malignancy. Algorithms for diagnostic evaluation and treatment of pediatric inflammatory bowel disease are presented. The specific psychosocial issues facing these patients are also discussed in this review as are the future goals of research in the complex problem of pediatric inflammatory bowel disease. PMID- 16718841 TI - Surgical perspectives in gastrointestinal disease: a study of quality of life outcomes in esophageal, pancreatic, colon, and rectal cancers. AB - Outcomes assessment in surgery traditionally has included data regarding peri operative mortality and morbidity, as well as long-term survival and recurrence in surgical oncology. However, quality of life (QOL) is another important patient related outcome measure. QOL data can be used to tailor treatment and improve clinical outcomes by detecting physical or psychological problems in patients that otherwise might be overlooked, but which have profound implications for the effective delivery of care. We review several well-validated QOL instruments developed specifically for use in patients with gastrointestinal malignancies, including esophageal, pancreatic and colorectal cancers, and conclude that QOL assessment routinely should be included in clinical trials of novel treatments. PMID- 16718842 TI - Evolution of surgical skills training. AB - Surgical training is changing: one hundred years of tradition is being challenged by legal and ethical concerns for patient safety, work hours restrictions, the cost of operating room time, and complications. Surgical simulation and skills training offers an opportunity to teach and practice advanced skills outside of the operating room environment before attempting them on living patients. Simulation training can be as straight forward as using real instruments and video equipment to manipulate simulated "tissue" in a box trainer. More advanced, virtual reality simulators are now available and ready for widespread use. Early systems have demonstrated their effectiveness and discriminative ability. Newer systems enable the development of comprehensive curricula and full procedural simulations. The Accreditation Council of Graduate Medical Education's (ACGME) has mandated the development of novel methods of training and evaluation. Surgical organizations are calling for methods to ensure the maintenance of skills, advance surgical training, and to credential surgeons as technically competent. Simulators in their current form have been demonstrated to improve the operating room performance of surgical residents. Development of standardized training curricula remains an urgent and important agenda, particularly for minimal invasive surgery. An innovative and progressive approach, borrowing experiences from the field of aviation, can provide the foundation for the next century of surgical training, ensuring the quality of the product. As the technology develops, the way we practice will continue to evolve, to the benefit of physicians and patients. PMID- 16718843 TI - Management of diverticular disease is changing. AB - Diverticular disease of the colon is primarily a disease of humans living in westernized and industrialized countries. Sixty percent of humans living in industrialized countries will develop colonic diverticula. It is rare before the age of 40, but more prone to complications when it occurs in the young. By age 80, over 65% of humans have colonic diverticula. The cause remains uncertain, but epidemiologic studies attribute it to dietary fiber deficiency. The cause of diverticulitis remains uncertain, but new observations and hypotheses suggest that it is due to chronic inflammation in the bowel wall. Standard medical therapies of bowel rest and antibiotics are still the recommended treatment. However, changing concepts and new therapies indicate that anti-inflammatory agents such as mesalamine and possibly probiotics may be helpful in shortening the course and perhaps preventing recurrences. Standard surgical treatment for perforation for severe acute disease has developed so that two-stage procedures are recommended. In addition, laparoscopic surgery has proven safe and may slowly become the technique of choice. PMID- 16718845 TI - Updates on esophageal and gastric cancers. AB - Esophageal and gastric cancers are both common and deadly. Patients present most often after disease progression and survival is therefore poor. Due to demographic variability and recent changes in disease incidence, much emphasis has been placed on studying risk factors for both esophageal and gastric cancers. However, with increasing understanding of these diseases, low survival rates persist and continued intensive studies are necessary to optimize treatment plans. This review article discusses updates in the evolving epidemiology, clinical presentation, risk factors, and diagnostic and treatment modalities of esophageal and gastric cancers. PMID- 16718844 TI - Role of calcium and other trace elements in the gastrointestinal physiology. AB - Calcium is an essential ion in both marine and terrestrial organisms, where it plays a crucial role in processes ranging from the formation and maintenance of the skeleton to the regulation of neuronal function. The Ca(2+) balance is maintained by three organ systems, including the gastrointestinal tract, bone and kidney. Since first being cloned in 1993 the Ca(2+)-sensing receptor has been expressed along the entire gastrointestinal tract, until now the exact function is only partly elucidated. As of this date it still remains to be determined if the Ca(2+)-sensing receptor is involved in calcium handling by the gastrointestinal tract. However, there are few studies showing physiological effects of the Ca(2+)-sensing receptor on gastric acid secretion and fluid transport in the colon. In addition, polyamines and amino acids have been shown to activate the Ca(2+)-sensing receptor and also act as allosteric modifiers to signal nutrient availability to intestinal epithelial cells. Activation of the colonic Ca(2+)-sensing receptor can abrogate cyclic nucleotide-mediated fluid secretion suggesting a role of the receptor in modifying secretory diarrheas like cholera. For many cell types changes in extracellular Ca(2+) concentration can switch the cellular behavior from proliferation to terminal differentiation or quiescence. As cancer remains predominantly a disease of disordered balance between proliferation, termination and apoptosis, disruption in the function of the Ca(2+)-sensing receptor may contribute to the progression of neoplastic disease. Loss of the growth suppressing effects of elevated extracellular Ca(2+) have been demonstrated in colon carcinoma, and have been correlated with changes in the level of CaSR expression. PMID- 16718846 TI - Update in management of mesenteric ischemia. AB - Mesenteric ischemia disorders are precipitated by a circulation insufficiency event that deprives one or several abdominal organs of adequate respiration to meet metabolic demands. Although mesenteric ischemia occurs infrequently, the mortality rate is from 60% to 100%, depending on the source of obstruction. The successful outcome is dependent upon a high index of suspicion and prompt management. We briefly review the pathophysiology and presentation of the various ischemic entities and review the current state of the art in diagnosis and treatment. Despite advances in both diagnosis and treatment, prompt diagnosis and supportive care remain critical for successful outcome. New imaging techniques, endovascular therapy and emerging research may improve our approach to this deadly condition. PMID- 16718847 TI - Surgical perspectives in peptic ulcer disease and gastritis. AB - For much of the twentieth century, surgery was frequently the solution for peptic ulcer disease. Our understanding of the pathophysiology of ulcers paralleled the development of potent pharmaceutical therapy. As the surgical world developed parietal cell vagotomy which would minimize the complications of surgery, patients failing medical therapy became rare. Emergent surgery for complicated peptic ulcers has not declined however. The development of proton pump inhibitors and the full understanding of the impact of H pylori has led to a trend towards minimalism in surgical therapy for complicated peptic ulcer disease. In addition to the changes in patient care, these developments have had an impact on the training of surgeons. This article outlines these trends and developments. PMID- 16718848 TI - Evolution in the treatment of metastatic colorectal carcinoma of the liver. AB - Metastatic colorectal cancer to the liver is associated with a uniform poor prognosis without treatment. Advances in therapy over the past decades have now allowed surgical resections of the liver to occur with a low morbidity and mortality. Improvements in chemotherapy regimes have paralleled technical improvements and now allow a new group of patients to become eligible for surgical resection. This chapter will review the recent advances in surgical and chemotherapeutic regimes in metastatic colorectal cancer to the liver. PMID- 16718849 TI - Ethical tensions in solid organ transplantation: the price of success. AB - Solid organ transplantation has rapidly developed into the therapy a choice for end-stage organ failure. The expansion of its use has resulted is a large deficiency in organ supply. To address this, the field of organ transplantation has attempted to develop new strategies that would increase the availability of organs for transplant. Some of these strategies include expansion of the donor pool by increasing the number of living donors or using deceased donor organs that may be marginal or "expanded". The intent is to bring life-saving therapy to individuals in need; however, much of this expansion has been brought forward without clear prospective guidelines. This article focuses on the current disparity between organ supply and demand, and how this has impacted the use of living donors and development of the "expanded donor" concept. PMID- 16718850 TI - Hepatic perfusion disorders: etiopathogenesis and related diseases. AB - In this article, we have reviewed the hepatic perfusion disorder (HPD), etiopathogenesis of HPD and corresponding diseases. Review of the literature was based on computer searches (PubMed, Index Medicus) and personal experiences. We considered HPD reflects perfusion differences due to redistribution of arterial blood flow among segments, subsegments and lobes of the liver. The plain CT scan findings of HPD manifests as triangular or wedge-shaped areas of low attenuation. On contrast-enhanced CT scan, HPD manifests multiple (or single) transient wedge shaped, rotundloid or irregular appearance, homogeneous hyperattenuation (in less cases, hypoattenuation) during the hepatic arterial phase (HAP) and isoattenuated or slightly hyperattenuated areas during the portal arterial phase. Dynamic enhanced magnetic resonance (MR) features are similar to enhanced CT scan. Angiographic findings include non-opacification of portal vein on portograms or wedge-shaped segmental staining in arterial and parenchymal phases on hepatic angiograms. The causes of HPD are arterioportal shunts (APS), intrahepatic vascular compressions and portal vein occlusion, steal phenomenon by hypervascular tumors, vascular variations and any other unknown reasons. It is very important for radiologists to be familiar with the various appearances of HPD to avoid false-positive diagnosis of pseudolesions and not to overestimate the extent of the disease. PMID- 16718851 TI - Effects of nucleus ambiguus and dorsal motor nuclei of vagus on gastric H(+) and HCO(3)(-) secretion in rats. AB - AIM: To determine the effects of electrical stimulation of nucleus ambiguus (NA) and dorsal motor nuclei of vagus (DMV) on gastric acid and bicarbonate secretion in rats. METHODS: NA and DMV in rats were electrically stimulated. Pylorus ligation or esophagus perfusion was used to collect the gastric secretion. The titratable H(+) quantum, H(+) concentration, HCO(3)(-) secretion quantum were measured. RESULTS: Electrical stimulation of NA had no effects on the volume of gastric juice, titratable acidity and acid concentration, but elicited a pronounced increase in the total bicarbonate. However, electrical stimulation of DMV significantly increased the titratable acidity, the volume of gastric juice and the acid concentration. Similarly, electrical stimulation of either NA or DMV decreased the respiratory frequency and sinus bradycardia. CONCLUSION: NA in rats can not control the secretion of gastric acid but the secretion of bicarbonate in gastric juice, while DMV controls the secretion of gastric acid. PMID- 16718852 TI - Direct ex vivo analysis of dendritic cells in patients with hepatocellular carcinoma. AB - AIM: To analyze the phenotype and function of dendritic cells (DC) from patients with hepatocellular carcinoma (HCC) in order to understand their role in this disease. METHODS: Myeloid dendritic cells were enumerated in peripheral blood of HCC patients. CD80, CD83, CD86 and HLA-DR expression on naive and stimulated myeloid dendritic cells from peripheral blood were analyzed. Myeloid dendritic cells were isolated from peripheral blood and their function was tested. Phagocytosis was analyzed using FITC-dextran beads, peptide specific stimulation, the capacity to stimulate allogeneic T cells and secretion of cytokines upon poly dI:dC was tested. RESULTS: Myeloid dendritic cells were reduced in patients with HCC. No differences in CD80, CD83, CD86 and HLA-DR expression were found on naive and stimulated myeloid dendritic cells from HCC patients and healthy controls. Normal phagocytosis or stimulation of peptide specific T cells was observed in contrast to an impaired allo-stimulatory capacity and a reduced IL-12 secretion. CONCLUSION: Impaired IL-12 production of mDCs in patients could lead to an impaired stimulatory capacity of naive T cells suggesting that IL-12 directed therapies may enhance tumor specific immune responses in HCC patients. PMID- 16718853 TI - Investigation of HER-2 codon 655 single nucleotide polymorphism frequency and c ErbB-2 protein expression alterations in gastric cancer patients. AB - AIM: To investigate both whether the risk of gastric cancer is associated with the Ile/Val single nucleotide polymorphism (SNP) of human epidermal growth factor receptor-2 (HER-2) transmembrane domain-coding region at codon 655 and the suggested existence of HER-2 expression in gastric cancer cases in a Turkish patient group. METHODS: Polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP) strategy was used to analyze the presence of HER-2 SNP at codon 655. c-erbB-2 expression pattern was analyzed by immunohistochemistry. The results were compared between gastric carcinoma group and chronic gastritis group, as well as between clinicopathological parameters and carcinoma. RESULTS: Results showed that Ile/Val genotype accounted for 20% within the Turkish gastric carcinoma group, and none in chronic gastritis group, and this genotyping was associated with stage IV gastric cancers (P=0.04). Positive membranous HER-2 immunoreactivity, on the other hand, accounted for 24% within the Turkish gastric carcinoma group and none from chronic gastritis cases; further, it was correlated with intestinal type carcinomas (P=0.007), and stage III-IV carcinomas (P=0.004). CONCLUSION: These observations imply that the tested HER-2 SNP may participate in the development and progression of gastric cancer. Thus, after confirming these results with large sample groups, HER-2 codon 655 SNP and/or c-erbB-2 overexpression may also be used as a poor prognostic indicator for gastric carcinomas. PMID- 16718854 TI - Ultrasonic interventional analgesia in pancreatic carcinoma with chemical destruction of celiac ganglion. AB - AIM: To detect the therapeutic effects of chemical destruction of celiac ganglion in patients with pancreatic carcinoma with intractable pain. METHODS: Ninety seven cases with advanced pancreatic carcinoma received chemical destruction of celiac ganglion-5 mL pure alcohol injection around celiac artery under ultrasonic guidance. The changes of visual analogue scale (VAS), serum substance P (Sub P), beta-endopeptide (beta-EP) and T-lymphocyte subtypes level were compared between pre- and post-therapy. RESULTS: Successful rate of puncture was 98.7%, with one failure. No serious complications such as traumatic pancreatitis, pancreatic fistula, abdominal cavity hemorrhage or peritoneal infection occurred. VAS, serum Sub P and beta-EP level significantly changed after treatment (8.0+/-2.3 vs 4.6+/ 2.1, 254.1+/-96.7 vs 182.4+/-77.6, 3.2+/-0.8 vs 8.8+/-2.1, P<0.01, P<0.05, P<0.01) with complete relief rate 54.2%, partial relief rate 21.9%, ineffective rate 12.5% and recurrent rate 10.7%. The T-lymphocyte subtypes level remarkably increased when compared with that of pre-therapy (46.7+/-3.7 vs 62.5+/-5.5, P<0.01). CONCLUSION: Our study suggests that chemical destruction of celiac ganglion under ultrasonic guidance is highly safe, and can evidently relieve cancer pain and improve the cellular immunity in patients with advanced pancreatic carcinoma. PMID- 16718855 TI - For better or worse? PMID- 16718856 TI - Psychiatric disorder is deeply entangled in a sociocultural web, of which gender is one strand. PMID- 16718857 TI - No matter how snazzy the software, mouse-driven decisions will always be subject to human error. PMID- 16718858 TI - Woman on the verge. PMID- 16718859 TI - Models for universal health care: how to fix a broken system. PMID- 16718860 TI - Rehabilitation of traumatic brain injury. Current guidelines and beyond. PMID- 16718861 TI - US Food and Drug Administration pushed to change. PMID- 16718862 TI - Court ends Colombia's abortion ban. PMID- 16718863 TI - Use of receiver operating characteristic (ROC) analysis to evaluate sequence matching. AB - In this paper, we borrow the idea of the receiver operating characteristic (ROC) from clinical medicine and demonstrate its application to sequence comparison. The ROC includes elements of both sensitivity and specificity, and is a quantitative measure of the usefulness of a diagnostic. The ROC is used in this work to investigate the effects of scoring table and gap penalties on database searches. Studies on three families of proteins, 4Fe-4S ferredoxins, lysR bacterial regulatory proteins, and bacterial RNA polymerase sigma-factors lead to the following conclusions: sequence families are quite idiosyncratic, but the best PAM distance for database searches using the Smith-Waterman method is somewhat larger than predicted by theoretical methods, about 200 PAM. The length independent gap penalty (gap initiation penalty) is quite important, but shows a broad peak at values of about 20-24. The length dependent gap penalty (gap extension penalty) is almost irrelevant suggesting that successful database searches rely only to a limited degree on gapped alignments. Taken together, these observations lead to the conclusion that the optimal conditions for alignments and database searches are not, and should not be expected to be, the same. PMID- 16718864 TI - Oligonucleotide frequencies in DNA follow a Yule distribution. AB - We show that ranked oligonucleotide frequencies in both protein-coding and non coding regions from several genomes fit poorly to the Zipf distribution, but that the same frequency data give excellent fit to the Yule distribution. The parameters of the Yule distribution for oligonucleotide frequencies in exons are the same (within error limits) as the parameters for introns. This precludes application of Yule or Zipf distribution of ranked oligonucleotide frequencies to annotating new genomic sequences. PMID- 16718865 TI - Chaotic interactions of self-replicating RNA. AB - A general system of high-order differential equations describing complex dynamics of replicating biomolecules is given. Symmetry relations and coordinate transformations of general replication systems leading to topologically equivalent systems are derived. Three chaotic attractors observed in Lotka Volterra equations of dimension n = 3 are shown to represent three cross-sections of one and the same chaotic regime. Also a fractal torus in a generalized three dimensional Lotka-Volterra Model has been linked to one of the chaotic attractors. The strange attractors are studied in the equivalent four-dimensional catalytic replicator network. The fractal torus has been examined in adapted Lotka-Volterra equations. Analytic expressions are derived for the Lyapunov exponents of the flow in the replicator system. Lyapunov spectra for different pathways into chaos has been calculated. In the generalized Lotka-Volterra system a second inner rest point--coexisting with (quasi)-periodic orbits--can be observed; with an abundance of different bifurcations. Pathways from chaotic tori, via quasi-periodic tori, via limit cycles, via multi-periodic orbits- emerging out of periodic doubling bifurcations--to "simple" chaotic attractors can be found. PMID- 16718866 TI - Providing HIV/AIDS care in a changing environment. Self-management and the chronic care model. PMID- 16718867 TI - A bio-psycho-social-spiritual approach to HIV and homelessness. PMID- 16718868 TI - Homeless in the age of AIDS. PMID- 16718870 TI - When your new diagnosis is with HIV. PMID- 16718869 TI - HIV as an emerging chronic disease: the challenge of "living with living". PMID- 16718871 TI - HIV therapy: a double whammy for heart disease and diabetes? PMID- 16718872 TI - Another diagnosis. Now what do I do? PMID- 16718873 TI - Personal perspective. Living with a multiple diagnosis. An interview with Body Positive board member Patricia Shelton. PMID- 16718874 TI - Managing fatigue in HIV/AIDS. PMID- 16718875 TI - Report from the 13th retrovirus conference. Integrase inhibitors on the way. PMID- 16718876 TI - Report from the 13th retrovirus conference. New data on TMC114 and TMC125. PMID- 16718877 TI - Report from the 13th retrovirus conference. Vicriviroc less effective than efavirenz in treatment-naive patients. PMID- 16718878 TI - Report from the 13th retrovirus conference. Treatment interruption, interrupted. PMID- 16718879 TI - Report from the 13th retrovirus conference. Should we be starting treatment earlier? PMID- 16718880 TI - Report from the 13th retrovirus conference. Metabolic complications and cardiovascular disease. PMID- 16718881 TI - Report from the 13th retrovirus conference. Renal tenofovir debate continues. PMID- 16718882 TI - Report from the 13th retrovirus conference. Extending the role of atazanavir. PMID- 16718883 TI - Report from the 13th retrovirus conference. Antiretroviral therapy in resource limited settings. PMID- 16718884 TI - Report from the 13th retrovirus conference. The new ABCs: antiretrovirals, barriers, and circumcision. PMID- 16718885 TI - Report from the 13th retrovirus conference. More on false-positive results in rapid HIV testing. PMID- 16718886 TI - Report from the 13th retrovirus conference. Expanding HIV testing. PMID- 16718887 TI - Antiretroviral rounds. A troublesome twosome: AIDS and persistent bacteremia. PMID- 16718888 TI - Truvada trials hold promise for new HIV prevention strategy. Once-a-day might keep HIV away. PMID- 16718889 TI - A completely new target for fighting HIV infection? Target involves DNA repair genes. PMID- 16718890 TI - Metabolic problems in children on ARTs found. Solution is finding balance in treatment. PMID- 16718892 TI - Therapy for family, patient improves adherence. Intervention designed to reduce drug abuse relapse. PMID- 16718891 TI - Research findings suggest new course against HIV. Nef protein plays a major role. PMID- 16718893 TI - DM program keeps AIDS patients out of hospital. One-on-one education is the key to success. PMID- 16718894 TI - HIV subtype efficient mortality predictor. Subtype D and AD lead to rapid deaths. PMID- 16718895 TI - Global. U.N.: doctor shortage harms AIDS fight. PMID- 16718897 TI - Attorney's HIV dementia fails to save client from penalty. PMID- 16718898 TI - GAO report shows Bush's global AIDS policy is failing. PMID- 16718896 TI - ADAP. Nearly 800 on U.S. waiting lists for AIDS drugs. PMID- 16718899 TI - Dismissal of an HIV-positive man's disability claim reversed. PMID- 16718900 TI - Discrimination. Inmate's ineffective counsel claim fails. PMID- 16718902 TI - Criminal transmission. HIV-positive Canadian man faces additional charges. PMID- 16718901 TI - High court to decide spousal HIV disclosure. PMID- 16718904 TI - Project Inform analysis: results of a "cure" study. PMID- 16718903 TI - Lessons in hope. PMID- 16718905 TI - Entry inhibitors: a race to the finish line. PMID- 16718906 TI - Understanding Medicare Part D. PMID- 16718907 TI - dD4FC development discontinued. PMID- 16718908 TI - Prevention. Officials nationwide urge proper needle disposal. PMID- 16718909 TI - Treatment. New York jails use computers to improve inmate HIV care. PMID- 16718911 TI - Global. Groups oppose restrictions on AIDS funding in court. PMID- 16718910 TI - Privacy. Judge blocks Kansas law to report sex under age 16. PMID- 16718912 TI - Global. WHO declares emergency against AIDS, TB, malaria. PMID- 16718913 TI - Fear of exposure to HIV falls short of damage recovery. PMID- 16718915 TI - Privacy. County must answer for broadcasting man's serostatus. PMID- 16718914 TI - Sentencing. False-positive HIV test fails to rescue man from guilty plea. PMID- 16718916 TI - Care and repair of your main artery. Early recognition and novel treatments are reducing the risks of vessel rupture. PMID- 16718917 TI - Blood pressure drugs linked with Alzheimer's protection. PMID- 16718918 TI - When cardiovascular drugs clash. Knowing how medications interact can protect you from risky side effects. PMID- 16718919 TI - The metabolic syndrome debate. What it means to you when experts disagree about a set of risk factors. PMID- 16718920 TI - Sex life linked to heart health. Erectile dysfunction may weigh in as heavily as family history, smoking. PMID- 16718921 TI - What is cardiomyopathy and what parts of the body does it affect besides the heart itself? What are its real causes, and what medications work best? Should I look at alternatives? PMID- 16718922 TI - Update on the recommendations for the routine use of pneumococcal conjugate vaccine for infants. An Advisory Committee Statement (ACS). PMID- 16718923 TI - Maternal depression: effects on parenting practices...and on the child when mother's depression improves. PMID- 16718924 TI - The September epidemic of asthma hospitalizations...and do antibiotics in infancy lead to asthma? PMID- 16718925 TI - Second FDA panel advises on stimulants. PMID- 16718926 TI - How parents view their child's weight. PMID- 16718927 TI - Postconditioning's protection of THSG on cardiac ischemia-reperfusion injury and mechanism. AB - 2,3,5,4'-tetra-hydroxystilbene-2-O-glucoside (THSG), the water-soluble active components extracted from dried tuber root of Polygonum multiflorum (Polygonaceae), can promote the release of nitric oxide (NO) from vascular endothelial cells and has strong antioxidation. The postconditioning's protection of THSG on cardiac ischemia-reperfusion injury and the mechanism were investigated. After reperfusion for 3 h following occlusion of rat left anterior descending coronary artery (LAD) for 30 min, SalphaT recovery speed, arrhythmia and cardiac infarct size were observed. The ischemic size and infarct size was identified by using Evans blue and TTC staining methods respectively. The results showed that the infarct size in THSG 7. 5 mg/kg postconditioning group was significantly decreased from 43.6% +/- 9.1% in mode group to 16.5% +/- 6.5% (P < 0.01). SalphaT recovery was quicker and the incidence of arrhythmia (55.6% vs 100%, P < 0.05) was significantly lower than in control group. The infarct size in THSG+glybenclamide group was greater than in THSG group, but equivalent to that in control group (46.8% +/- 9.8% vs 43.6% +/- 9.1%, P > 0.05), SalphaT recovery speed slower and the incidence of arrhythmia also lower (33.3% vs 100%, P < 0.01), suggesting that glybenclamide could abolish the effects of THSG postconditioning reducing the cardiac infart size. It was concluded that THSG administration before reperfusion could effectively alleviate the cardiac reperfusion injury and possessed the postconditioning effects of reducing cardiac infarct size, which might be related with the K(ATP) channel opening. PMID- 16718928 TI - Inhibition of the VEGF expression and cell growth in hepatocellular carcinoma by blocking HIF-1alpha and Smad3 binding site in VEGF promoter. AB - In order to investigate the inhibitory effects on the vascular endothelial growth factor (VEGF) expression and cell growth in hapatocellular carcinoma (HCC) by blocking HIF-1alpha and Smad3 binding site in the VEGF promoter, antisense oligodeoxynucleotides (ASODN) were designed to block HIF-1alpha and Smad3 binding site in the VEGF promoter. Different concentrations of ASODN and ODN were transfected into HCC cells respectively. The expression of VEGF mRNA and protein was detected by SABC, Western blot and RT-PCR techniques and the inhibitory effects on the expression of VEGF and cell growth of the HCC cells stimulated by the supernatants were determined by using MTT method. Immunohistochestry revealed that after co-inoculation of hepatocellular carcinoma cells with different concentrations of ODN and ASODN for 48 h, there was no significant difference in the expression of VEGF protein between ODN group and control group (P > 0.05), but there was significant difference between ASODN group and control group (P < 0.05). At a concentration of 10 micromol/L ASODN, the difference was very significant (P < 0.01). Western blot and RT-PCR revealed that, after treatment for 48 h at a concentration of 10 micromol/L. the integral gray levels and RNA odds were 59743.2 +/- 10412.5 and 0.783 +/- 0.032 in ODN group, and 38694.5 +/- 10925.1 and 0.468 +/- 0.015 in ASODN group, respectively, with the difference being very significant (P < 0.01). Antisense ODN could inhibit the growth of HCC cells in a concentration-dependent manner. It was concluded that anti-gene technique of aiming at HIF-1alpha action site in the VEGF promoter could suppress the VEGF expression and inhibit HCC cell growth, and it is promising that anti gene technique works as a new gene therapeutic tool for anti-angiogenesis of HCC. PMID- 16718929 TI - Up-regulation of niacinamide in intervertebral disc aggrecan in vitro. AB - The regulatory effects of niacinamide (Nia) on intervertebral disc (IVD) aggrecan in vitro was investigated. Chiba's 10 ng/mL interleukin-1 (IL-1)-induced rabbit IVD degeneration model in vitro was established. 0.5, 0.25 and 0.05 mg/mL Nia was added to normal and degenerated IVDs for intervention. On the first and second week after intervention, safranin O-fast green staining intensity and glycosaminoglycan (GS) content were measured. The expression of aggrecan core protein was detected by RT-PCR. The results showed: (1) After treatment with 0.5 mg/mL Nia for one week, the GS content in nucleus pulposus (NP) was increased by 44.8% as compared with control group (P < 0 01); The GS content in IL-1 induction groups was increased with the increase of Nia concentrations: After treatment with 0.5 mg/mL for one week, the GS content in NP was increased by 68.3% as compared with control group (P < 0.01). After two weeks, GS content in NP and fibrous rings was still higher than in control group at the same period (P < 0.01) and untreated group (P < 0.01). (2) Safranin O-fast green staining revealed that with the increase of Nia concentrations, staining density in NP and fibrous rings was increased and histological structure damage to IVDs by IL-1beta was alleviated. (3) RT-PCR showed that the expression of core protein gene in IL 1beta-induced degenerated IVDS was increased with the increase of Nia concentrations. It was concluded that under conditions in vitro, Nia could up regulate the expression of aggrecan in IVDs and protect IVDs from IL-1beta induced degeneration at least partially, which offers a potential choice for IVD degeneration clinical therapy. PMID- 16718930 TI - [University training for coordinators]. AB - We need an adequate quality level of information between the building design and manufacturing process for Health and Safety in the work site. A fully-developed communication system is needed, for H&S Coordinators, in order to integrate alphanumerical and graphical protocols. The Aim is: (1) to organise information on the building process, keeping communications in function of everyone's needs; (2) to carry out an audit process ensuring that all levels of contractors personnel are implementing the Project of Health and Safety Management Plan effectively; (3) To make all that with a Program Interface relatively simple. The synergy between Safety Manager and Designers generates a system output integrating all sorts of alphanumerical, graphical information and prescriptions. This system output has several targets: (1) to integrate Design and H&S in a specific Data-Base; (2) to define the preconditions of the work phases; (3) to develop the Work Program of Enterprise in function of the Health and Safety Plan in compliance with the contractor site-specific plan; (4) to develop a program process for the audit; (5) to review the contractor's Project for H & S; (6) to manage responsibility; (7) to keep corrective action. PMID- 16718931 TI - [A 35-year-old man with recurrent mouth, periurethral, and scrotal ulcers]. PMID- 16718932 TI - A simple regression method for mapping quantitative trait loci in line crosses using flanking markers. AB - The use of flanking marker methods has proved to be a powerful tool for the mapping of quantitative trait loci (QTL) in the segregating generations derived from crosses between inbred lines. Methods to analyse these data, based on maximum-likelihood, have been developed and provide good estimates of QTL effects in some situations. Maximum-likelihood methods are, however, relatively complex and can be computationally slow. In this paper we develop methods for mapping QTL based on multiple regression which can be applied using any general statistical package. We use the example of mapping in an F(2) population and show that these regression methods produce very similar results to those obtained using maximum likelihood. The relative simplicity of the regression methods means that models with more than a single QTL can be explored and we give examples of two lined loci and of two interacting loci. Other models, for example with more than two QTL, with environmental fixed effects, with between family variance or for threshold traits, could be fitted in a similar way. The ease, speed of application and generality of regression methods for flanking marker analysis, and the good estimates they obtain, suggest that they should provide the method of choice for the analysis of QTL mapping data from inbred line crosses. PMID- 16718934 TI - Spondylodiscitis after facet joint steroid injection: a case report and review of the literature. AB - Spondylodiscitis is a rare complication after facet joint steroid injection. This article presents a 78-y-old male with chronic back pain and facet joint arthritis who developed Pseudomonas aeruginosa L2-L3 and L3-L4 spondylodiscitis after computed tomography-guided facet joint steroid injection. Magnetic resonance imaging, fine needle aspiration and cultures confirmed the diagnosis. The patient was treated with intravenous administration of amikacin, and imipenem plus cilastatin for 4 weeks, followed by oral administration of ciprofloxacin for another 10 weeks. Sterile preparation prior to facet joint steroid injection should be stressed. To the best of our knowledge, this is the only reported case of iatrogenic spondylodiscitis after facet joint steroid injection. This iatrogenic complication should be considered in the risk-benefit analysis of facet joint diagnostic or therapeutic injections. PMID- 16718933 TI - Combination therapy with daptomycin, vancomycin, and rifampin for recurrent, severe bone and prosthetic joint infections involving methicillin-resistant Staphylococcus aureus. AB - Methicillin-resistant Staphylococcus aureus (MRSA) infections are commonly treated with vancomycin (VAN) or another glycopeptide antibiotic. However, when vancomycin fails or infections recur, there are few other therapeutic options. Presented here are 2 cases where a novel combination of daptomycin, vancomycin, and rifampin resolved recurrent MRSA bone and prosthetic joint functions. PMID- 16718935 TI - Isolated tuberculosis of the vastus lateralis muscle: a case report. AB - Tuberculosis is 1 of the most deadly diseases in the world. With an increase in the incidence of HIV worldwide, tubercular infections at unusual sites are being reported. Skeletal muscle tuberculosis without bony involvement is an extremely rare presentation of tuberculosis. A case of isolated tuberculosis of the vastus lateralis muscle without any evident primary focus in a 30-y-old immunocompetent female is presented. PMID- 16718936 TI - Severe disseminated tuberculosis in a 4-month-old infant initially presenting with multiform cutaneous lesions. AB - A 4-month-old female infant presented with 1 month history of horizontal nystagmus and discrete multiform skin lesions. The patient was initially diagnosed with congenital nystagmus and staphylococcal skin infection not responding to antimicrobial agents. The development of severe systemic symptoms led to extensive investigations and the diagnosis of disseminated tuberculosis. Mycobacterium tuberculosis (MTB) was isolated from the skin and cerebrospinal fluid. The patient was treated with isoniazid, rifampicin, pyrizinamide, streptomycin and dexamethasone. Skin lesions resolved completely but severe neurological deficits persisted. PMID- 16718937 TI - Multifocal skeletal tuberculosis with Rhodotorula minuta co-infection. AB - Multifocal skeletal tuberculosis comprises lesions occurring at 2 or more locations of the skeleton. We present an atypical case of multifocal skeletal tuberculosis coexisting with Rhodotorula minuta infection in a non immunocompromized host. The advantages of computed tomography-guided biopsy in establishing the diagnosis and thus influencing treatment are also discussed. PMID- 16718938 TI - [The 18th spring meeting of the Japanese Society of Allergology. Tokyo, Japan. May 30-31, June 1, 2006. Abstracts]. PMID- 16718939 TI - Thymoma with immunodeficiency (Good's syndrome): review of the literature apropos three cases. AB - Good's syndrome is the association of thymoma with immunodeficiency, characterized by hypogammaglobulinaemia, depleted B-cells, diminished T-cells and inversion of the CD4/CD8 ratio. The initial clinical presentation is either with a mass lesion-thymoma that is usually benign, or with recurrent infections due to immunodeficiency. Thymectomy usually favourably affects associated conditions, such as pure red cell aplasia, but does not improve hypogammaglobulinaemia, thus the patient remains dependent on intravenous immune globulin and prone to infections. Infections usually affect the respiratory and/or the gastrointestinal tract. Common respiratory, opportunistic, and eventually life-threatening infections may occur. Moreover, patients with Good's syndrome may present other haematological conditions. We report 3 cases with long follow-up, sharing some common manifestations of the syndrome, but also showing unique features. The principal features of this rare syndrome are further discussed. PMID- 16718940 TI - [The 90th Congress of the Japanese Society of Legal Medicine. Fukuoka, Japan. April 26-28, 2006. Abstracts]. PMID- 16718942 TI - [The 49th annual meeting of the Japanese Society of Nephrology. Tokyo, Japan. June 14-16, 2006. Abstracts]. PMID- 16718941 TI - Lamotrigine-induced toxic epidermal necrolysis in three patients treated for bipolar disorder. AB - Exfoliative dermatitis, including Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN), is a mild-to-life-threatening process that has been described after exposure to many antiepileptic drugs. The increased use of antiepileptic drugs for treatment of bipolar disorder and neurologic disorders has extended the risk of exfoliative disorder to this population of patients, and these patients and their health care providers may not be familiar with the risks involved with these drugs. We describe the cases of a 28-year-old woman with bipolar 1 disorder initially treated with lamotrigine, and two adolescent girls with bipolar 2 disorder treated with lamotrigine after poor responses to other drug regimens. In all three patients, rashes progressed to toxic epidermal necrolysis in spite of treatment with corticosteroids at their local hospitals; thus, they were transferred to our burn treatment center. Response to early corticosteroid treatment in suppressing progression of exfoliation was variable in these patients. Ultimately, two of the three required ventilatory support; their conditions improved within 8-32 days of treatment, and they were discharged from the hospital. Case reports of lamotrigine-induced exfoliative disorder in patients with bipolar disorder have been published. However, these three patients were admitted to our burn treatment center within a 12-month period. Our institution admits approximately 10-12 patients with TEN/year, and the increased use of lamotrigine for treatment of bipolar disorder is likely to result in more patients with TEN. Therefore, health care professionals need to be aware of the early signs and symptoms of exfoliative dermatotoxicity when treating patients with lamotrigine. PMID- 16718943 TI - Severe serum sickness reaction to oral and intramuscular penicillin. AB - Serum sickness is a type III hypersensitivity reaction mediated by immune complex deposition with subsequent complement activation, small-vessel vasculitis, and tissue inflammation. Although the overall incidence of serum sickness is declining because of decreased use of heterologous sera and improved vaccinations, rare sporadic cases of serum sickness from nonprotein drugs such as penicillins continue to occur. Drug-induced serum sickness is usually self limited, with symptoms lasting only 1-2 weeks before resolving. We report an unusual case of a severe and prolonged serum sickness reaction that occurred after exposure to an intramuscular penicillin depot injection (probable relationship by Naranjo score) and discuss how pharmacokinetics may have played a role. Clinicians should be familiar with serum sickness reactions particularly as they relate to long-acting penicillin preparations. Accurate diagnosis in conjunction with cessation of drug exposure and prompt initiation of antiinflammatory treatment with corticosteroids can produce complete recovery PMID- 16718944 TI - American Association of Clinical Endocrinologists Medical Guidelines for Clinical Practice for the diagnosis and treatment of hypertension. PMID- 16718945 TI - Lepirudin anticoagulation for heparin-induced thrombocytopenia after cardiac surgery in a pediatric patient. AB - A 21-month-old boy received anticoagulation with lepirudin for heparin-induced thrombocytopenia (HIT) after undergoing cardiac surgery and receiving extracorporeal membrane oxygenation (ECMO). This report illustrates the significance of HIT in pediatric patients after cardiac surgery and the successful administration of lepirudin in this setting. To our knowledge, this is the first published report of lepirudin administered to treat HIT in a child after cardiac surgery and ECMO. Although guidelines exist that suggest the potential administration of lepirudin as treatment for children with HIT, further studies are needed to determine the safest yet most effective dosage for this population. PMID- 16718946 TI - Recurrent acute nitrofurantoin-induced pulmonary toxicity. AB - Nitrofurantoin may be used for prophylaxis of recurrent urinary tract infections in women; however, this agent has been associated with acute, subacute, and chronic pulmonary adverse reactions. The acute reaction occurs in about 1/5,000 women after their first exposure to the drug. We report the occurrence of two successive, highly probable (by Naranjo score) nitrofurantoin-induced acute pulmonary reactions in the same patient. On day 4 of prophylaxis with nitrofurantoin 100 mg/day (to prevent urinary tract infections), the patient developed intense substernal pain and pressure. On day 8, she experienced intense substernal burning. She went to the emergency department, where she vomited and was hypotensive and febrile. Her chest radiograph showed bilateral infiltrates. The patient was diagnosed with pneumonia and was prescribed levofloxacin; she stopped taking the nitrofurantoin. Her symptoms subsided over the next 3 days. Ten days later, the patient restarted the nitrofurantoin, and she returned to the emergency department after again experiencing sudden intense substernal burning, nausea, vomiting, shivering, and weakness. Nitrofurantoin was discontinued; her symptoms resolved quickly, and prophylaxis with trimethoprim-sulfamethoxazole was begun. After 1 year, the patient had experienced no further pulmonary symptoms or urinary tract infections. Drug toxicity must be considered in patients who develop pulmonary symptoms while taking nitrofurantoin. Symptoms are often misdiagnosed as other ailments, potentially subjecting patients to unnecessary treatments and delaying discontinuation of nitrofurantoin. Patients should be advised to contact a physician if breathing difficulties or unusual symptoms develop while taking nitrofurantoin, as this could result in earlier recognition of this drug reaction. PMID- 16718947 TI - Probable interaction between tolterodine and warfarin. AB - 53-year-old woman with a history of deep vein thrombosis and pulmonary embolism was receiving warfarin to prevent thromboembolic complications; her international normalized ratio (INR) had been stable for 1 month. Extended-release tolterodine 4 mg/day was then prescribed to manage overactive bladder. On her next anticoagulation clinic visit, the patient's INR had increased, although the dosage of warfarin had been reduced when the tolterodine had been prescribed. Due to the absence of other contributing factors and the temporal relationship between tolterodine and prolonged INR, the event was determined to be a probable drug interaction. When patients are prescribed tolterodine and warfarin concurrently, clinicians should monitor INR carefully, and a reduction in warfarin dosage may be required. PMID- 16718948 TI - Mucins in the diagnosis and differential diagnosis of pancreatic cystic neoplasms: report of 40 cases. PMID- 16718949 TI - Quality of care provided to patients with diabetes mellitus in Puerto Rico; managed care versus fee-for-service experience. AB - OBJECTIVE: To evaluate and compare the quality of diabetes care in a large managed care system and fee-for-service payment system in Puerto Rico. METHODS: This retrospective cross-sectional study assessed the adherence to standards of diabetes care in 1,687,202 subjects--226,210 from a fee-for-service population and 1,460,992 from a managed care group. Patients with diabetes mellitus were identified from insurance claims reports. Type of health-care provider, service location, number of visits, and laboratory utilization were also assessed. RESULTS: From the analysis, we identified 90,616 patients with diabetes (5.4% of the overall study group). Of these, 66,587 (73.5%) were found to have at least one encounter with a physician in a medical visit. Of the 66,586 patients with diabetes who visited a physician, only 4% were treated by an endocrinologist. General laboratory utilization was 34% for the entire population of patients with diabetes studied. In the group of patients with documented laboratory tests, 93% had a documented fasting blood glucose test; in contrast, hemoglobin A lc testing was performed in only 9% of the patients. The fee-for-service group had a higher rate of visits to medical specialists and general laboratory utilization, whereas the managed care group had a higher rate of hospital admissions and emergency department visits. CONCLUSION: The quality of diabetes management and the subsequent outcomes are related to patient and health-care provider adherence to standards of care. In this analysis, we found that patients and physicians are responsible for low compliance with recognized standards of diabetes care in Puerto Rico. The lack of adequate management will lead to increased mortality, development and severity of chronic complications, and increased emergency department utilization. Therefore, health-care providers and payers should find ways to achieve more effective promotion of adherence to accepted standards of care for patients with diabetes. PMID- 16718950 TI - Pilot measurements of ELF contact currents in some electric utility occupations. AB - Contact currents from touching objects with different voltages can produce electric fields within the body that produce neurological and other biological effects. To begin measuring these exposures among electric utility workers, a new contact current meter (CCM) was tested in a pilot study at Southern California Edison. The CCM was worn for 82 full-shift measurements by 76 volunteers from eight occupations who did not work directly with energized electrical equipment. The volunteers were exposed to an average of 285.8 contact current events above the meter's 1-microA threshold, but most of these were electrostatic spark discharges. Fourteen employees experienced an average of 135.1 contact currents events whose primary frequency was 60 Hz. Using a circuit model of the human body, the average contact currents going from arm to arm was 9.8 microA (maximum = 178.0 microA), and the average going down the torso was 25.5 microA (maximum = 662.0). The maximum exposures were experienced by a technical support employee working in a substation. All measurements in this pilot study were below the 3000 microA maximum permissible exposure for contact currents set by the Institute of Electrical and Electronic Engineers (IEEE). Combining these current measurements with the results of high-resolution dosimetry, the internal electric fields averaged an estimated 1.7 mV/m in the heart (maximum = 21.0 mV/m), and 1.9 mV/m in the hematopoietic bone marrow in the torso (maximum = 56.5 mV/m). These internal electric fields from contact currents are below the basic restriction of 943 mV/m in the IEEE exposure standards but are above 1 mV/m, a level where biological effects have been often reported in laboratory studies. Safety concerns limited the measurements to de-energized equipment, so we did not obtain data on work in energized high-voltage environments, the most likely sources of high contact currents. This pilot study identified other improvements to the contact current meter that would make it better able to measure exposures in future health studies. PMID- 16718951 TI - House dust collection efficiency of the high volume small surface sampler on worn carpets. AB - The High Volume Small Surface Sampler (HVS3) is a dust-sampling vacuum that allows for set airflow and back pressure during sampling, increasing precision. Total dust collection efficiency of the HVS3 has been evaluated only on new carpets-not worn carpets. We performed a factorial study to assess the impact of carpet wear, dust deposition level, carpet type, and relative humidity during sampling on HVS3 collection efficiency. House dust was aerosolized in a 1-m3 exposure chamber and allowed to settle on test carpets and reference filters. Dust was embedded into the carpets and later extracted with the HVS3 under controlled environmental conditions according to established protocols. Overall collection efficiency was high, 88.3%. Collection efficiency was significantly higher at low relative humidity levels (30%) relative to high (75%) (p = < 0.0001), though differentially between cut-pile and closed-loop carpets. Collection efficiency of carpets with high wear was significantly lower than those with midlevel wear (p = 0.01). These results demonstrate that the design of the HVS3 partially corrects for differences in dust load and carpet type. However, collection efficiency of the HVS3 is affected by high levels of carpet wear and ambient humidity during sampling. PMID- 16718953 TI - Statement on travellers and sexually transmitted infections. An Advisory Committee Statement (ACS). PMID- 16718952 TI - Jules Tinel (1879-1952) and Paul Hoffman (1884-1962). PMID- 16718954 TI - [Early Orthodontic Treatments: Do the Results Justify All the Effort? 78th Scientific Meeting of the French Society of Dento-Facial Orthopedics. Brussels, 25-28 May 2006]. PMID- 16718955 TI - Design and culture innovations combine at Baldrige winner. PMID- 16718956 TI - [Proceedings from the scientific meeting on medical opinioning: Dilemmas and problems of medical opinion. Poznan, 16-18 June 2005]. PMID- 16718957 TI - Abortion, intimacy, and the duty to gestate. AB - In this article, I urge that mainstream discussions of abortion are dissatisfying in large part because they proceed in polite abstraction from the distinctive circumstances and meanings of gestation. Such discussions, in fact, apply to abortion conceptual tools that were designed on the premiss that people are physically demarcated, even as gestation is marked by a thorough-going intertwinement. We cannot fully appreciate what is normatively at stake with legally forcing continued gestation, or again how to discuss moral responsibilities to continue gestating, until we appreciate in their own terms the goods and evils distinctive of gestational connection. To underscore the need to explore further the meanings of gestation, I provide two examples of the difference it might make to legal and moral discussions of abortion if we appreciate more fully that gestation is an intimacy. PMID- 16718958 TI - Contra-contraception: a growing number of conservatives see birth control as part of an ailing culture that overemphasizes sex and devalues human life. Is this the beginning of the next culture war? PMID- 16718959 TI - Doctors object as drug makers learn who's prescribing what. PMID- 16718960 TI - Panel faults Pfizer in '96 clinical trial in Nigeria. PMID- 16718961 TI - Disgraced Korean cloning scientist indicted. PMID- 16718962 TI - Colombian court legalizes some abortions. PMID- 16718963 TI - Screening for vertebral fracture with dual x-ray absorptiometry. PMID- 16718964 TI - Laparoscopic gastric bypass surgery for morbid obesity. PMID- 16718965 TI - Full-field digital mammography. PMID- 16718966 TI - Exhaled nitric oxide monitoring as a guide to treatment decisions in chronic asthma. PMID- 16718967 TI - Pulmonary vein isolation for treatment of atrial fibrillation. PMID- 16718968 TI - Special report: wireless esophageal pH monitoring. PMID- 16718970 TI - The staff of the future. PMID- 16718969 TI - Computer-aided detection with full-field digital mammography. PMID- 16718972 TI - Campaigning for continuing care. PMID- 16718973 TI - Progress on elder abuse. PMID- 16718974 TI - Guided by the light. PMID- 16718975 TI - Living well in later life. PMID- 16718976 TI - Will Wanless work? PMID- 16718977 TI - Ask the experts. Improving care for older people in hospital. PMID- 16718978 TI - Without prejudice: results and realisation of a training needs audit in nursing homes. AB - An analysis of training needs among registered nurses and carers in nursing homes will be used to form education action plans that will foster a learning culture and, ultimately, improve patient care PMID- 16718980 TI - Behind the scenes. PMID- 16718979 TI - Pressure ulcer prevention in older people who sit for long periods. AB - Pressure ulcers cause distress to patients and give health professionals cause for concern because of the dramatic effects on patients' quality of life, as well as the implications for resources (Hampton and Collins 2001). By understanding the changes in tissue physiology that take place and the complications of sitting, nurses can make pressure ulcer prevention part of their practice. PMID- 16718983 TI - Ginny Storey. PMID- 16718984 TI - What the future holds. PMID- 16718985 TI - Reflecting on change. PMID- 16718986 TI - Transforming the organisation of care. PMID- 16718987 TI - Using change in nursing practice: a case study approach. PMID- 16718988 TI - Redesigning patient services. PMID- 16718989 TI - Leading in teams: part 2. PMID- 16718990 TI - Effectiveness of an oral hygiene chew to reduce dental deposits in small breed dogs. AB - Over a period of 4-months, feeding small breed dogs with a regular dry diet supplemented with a daily oral hygiene chew resulted in a significant reduction in plaque deposition (17.3%) and calculus accumulation (45.8%) compared with dogs fed the dry diet alone. Daily feeding of a specifically designed oral hygiene chew that reduces plaque and calculus accumulation may play a role in the severity of canine periodontal disease. PMID- 16718991 TI - Feline caries in two cats from a 13th century archeological excavation. AB - Mandibles of two cats containing carious lesions were discovered among the previously published findings of feline dental resorptive lesions from materials examined at an archaeological museum. These lesions were too small to be noted on radiographs, and consisted of two inconspicuous enamel lesions in a mandibular left first molar tooth (309), a clinically visible white spot area containing an enamel lesion in a mandibular left fourth premolar tooth (308), and a root surface caries in the 308 of a different specimen. Histologic examination using special stains and polarized light revealed both initial and early initial stage enamel caries, as well as root surface caries. Knoop hardness measurements confirmed these findings, considered the first documented cases of feline caries. PMID- 16718992 TI - Management of multiple dental infections in a dog with diabetes mellitus. AB - A dog with diabetes mellitus was treated for multiple dental fractures and refractory hyperglycemia. Five teeth were found to have endodontic infections. Three extractions and two root canal procedures were performed to resolve these infections, resulting in improved regulation of blood glucose levels. Treatment planning included special considerations in management of diabetes in the anesthetic and perioperative periods. PMID- 16718993 TI - Full-thickness labial advancement flap following mast cell tumor excision. PMID- 16718994 TI - Pathophysiology and management of surgical and chronic oral pain in dogs and cats. PMID- 16718995 TI - Surgical endodontic therapy of the mandibular canine tooth. PMID- 16718996 TI - [Special project "improvement of the Japanese Society of Allergology and Medical Treatment for Allergic Disease"]. PMID- 16718997 TI - [IGE-FCepsilonRI-mast cell paradigm in allergy]. PMID- 16718998 TI - [Management of food allergy (Food allergy management 2005 by National Food Allergy Research Group)]. PMID- 16718999 TI - [Prognosis of bronchial asthma in adults]. AB - BACKGROUND: The prognosis of adult asthma, whether a long-term remission is available, is still unknown. In this paper we investigated the prognosis of adult asthma patients. METHODS We sent a questionnaire by mail to 1168 patients who had been taken care in our clinic until 1990-1992, but disappeared afterward. Those patients were asked their present status of clinical condition of asthma including symptoms, medication, etc. RESULTS: Delivery of mail was failed in 370 patients because of changed address. 430 of 798 patients replied the mail and 86 patients out of 430 patients were in remission state with no symptom without any medications. The characterictis of these patients in remission are early hospital visit after developing asthma, mild in severity, mild in obstructive lung function and mild in bronchial hypersensitivity to acetylcholine at the first visit hospital. CONCLUSION: We concluded that some of adult asthmatic patients might become in clinical remission. PMID- 16719000 TI - [Immedeate and delayed-type reactivity to fungi and effects of antifungal drugs on atopic dermatitis]. AB - OBJECTIVE: To investigate immediate and delayed-type reactivity for fungi in atopic dermatitis (AD) patients and the effect of antifungal therapy. METHODS: We examined immediate and delayed-type reactivity in AD patients for Candida albicans and Malassezia furfur by skin prick test (SPT), and estimated the effect of amphotericin B (AMPH) and itraconazole (ITCZ). RESULTS: Twenty eight of 40 patients showed positive immediate-type reaction and 10 of 27 patients did delayed-type reaction for Candida albicans. As for Malassezia furfur, positive immediate-type reaction was shown in 30 of 40 patients and positive delayed-type reaction did in 4 of 27 patients. The RAST score of specific IgE to Candida albicans was low in the patients with positive delayed-type skin reaction for Candida albicans, while the score was high in the patients with the negative delayed-type skin reaction. Both of AMPH and ITCZ were effective to the patients with positive immediate-type reaction for Candida albicans in SPT. The skin reaction for Malassezia furfur was stronger in the patients treated with ITCZ effectively than in the patients treated not effectively. In addition, ITCZ was effective in all patients except one, who showed positive reaction for Malassezia furfur accompanied with negative reaction for Candida albicans in SPT. CONCLUSION: Fungal allergy is one of the aggravation factors of AD, and SPT is useful to evaluate fungal allergy and to choose effective antifungal therapy. PMID- 16719001 TI - [Analysis of the prevalence of subtypes of urticaria and angioedema]. AB - BACKGROUND: The prevalence of various subtypes of urticaria in Japan has not been analyzed. METHODS: We have classified 260 (105 males and 155 females) out patients with urticaria based on the classification in Guidelines for the diagnosis and treatment of urticaria and angioedema" prepared by the Japanese Dermatological Association with slight modification. RESULTS: The urticaria in 200 out of 260 (76.9%) patients was idiopathic, and 101 (38.8%) patients showed more than one type of urticaria or angioedema. Among 146 patients with chronic urticaria, 90 (61.6%) were complicated by other types of urticaria; 59 (40.4%) by factitia, 29 (19.9%) by angioedema, and 12 (8.2%) by other types of urticaria. Deep pressure urticaria has been assumed as a rare type of urticaria in Japan, but was diagnosed in 5 patients with other types of urticaria. CONCLUSION: Most of the subtypes of urticaria could be diagnosed mainly by history and physical examinations. Correct diagnosis of the subtypes of urticaria should be the basis of better understanding and the treatment of this disease. PMID- 16719002 TI - [Feature of food allergy developed during infancy (1)--relationship between infantile atopic dermatitis and food allergy]. AB - BACKGROUND: Most of food allergy (FA) cases during childhood start as infantile atopic dermatitis (AD) at the ages of a few months old. We tried to clarify the association between infantile AD and FA during infancy. METHODS: We analyzed relationship between AD and FA during infancy among patients with 208 cases, who had visited our outpatient clinic with chief complaint of "eczema" from 1998 to 2000. RESULTS: Among 208 cases, 148 cases (71%) were diagnosed as infantile AD, moreover 109 cases (74%) were diagnosed as FA among infantile AD. The most frequent food antigens among infantile AD were egg (72.3%), cow's milk (39.9%), wheat (12.2%) and soybean (7.4%), respectively, in addition to these food antigens, food allergy was widely recognized against peanuts, sesame, meats, buckwheat, fishes and potato. In terms of food antigen, 44 cases with single food allergy against egg were seen out of 46 single allergy cases, whereas 36 cases with double food allergy against both egg and cow's milk were seen in 63 multiple food allergy cases. Although the value of antigen specific IgE against egg and cow's milk was recognized for the diagnosis of food allergy during infancy, even cases with negative IgE against those foods were proved to be food allergy by food elimination and provocation tests. In contrast to egg and cow's milk, positive IgE against rice, soybean, and wheat did not always correlate with the results of the diagnosis of food allergy. Concerning risk factors of AD, family history of any allergy diseases and passive smoking were recognized in comparison with infantile eczema. Neither the nutrition method nor incomplete elimination of diet during pregnancy and lactation had anything to do with the development of AD. CONCLUSION: When infantile AD cases were not improved by environmental control, skin care and application of steroid ointment, it would be important for doctors to think of the possibility of FA. PMID- 16719004 TI - Patient-led reform: from rhetoric to reality. PMID- 16719003 TI - Self-regulation--your call. PMID- 16719005 TI - Promoting public participation. PMID- 16719006 TI - A framework for success. PMID- 16719007 TI - The breastfeeding network: ripples in the pond. PMID- 16719008 TI - Express yourself and make a difference! PMID- 16719009 TI - Reach out to all mothers. PMID- 16719010 TI - Maternal pushing: to coach or not to coach. PMID- 16719011 TI - Controlling the capricious cervix (2). PMID- 16719012 TI - Midwifery basics: complications (7). Eclampsia. PMID- 16719014 TI - Evidence-based practice and critical care nursing. PMID- 16719013 TI - Time to get out of bed. PMID- 16719015 TI - You're fired! Sir Alan Sugar's response to the case made by NHS financial managers? PMID- 16719016 TI - Partnership working between a higher education institution and NHS TRUSTS: developing an acute and critical care module. AB - The underpinning philosophy of preparing nurses to work within a modern National Health Service (NHS) is to ensure patient safety and quality of clinical care. There is increasing recognition that post-registration education should become more clinically focused and adopt a more work-based approach. Critical to the success of such an approach is partnership working between education providers and NHS Trusts. Changes in the delivery of acute care in the NHS have resulted in an increased number of highly dependent patients in both critical and non critical care areas. This article describes a partnership approach between a Higher Education Institution (HEI) and two NHS Trusts to deliver a degree level module to address the subsequent educational implications of these changes. This approach ensured module credibility and clinical focus. Student feedback confirms the clinical relevance of the module. Staff from the NHS had the opportunity to develop teaching skills by preparing teaching materials, delivering teaching sessions and receiving feedback on these. Areas of good practice were identified and there was sharing of skills, knowledge and expertise between both institutions. Joint working can enhance clinical relevance of educational provision and ensure consistency between theory and practice. In addition, there were a number of wider benefits to both institutions that resulted from this initiative. PMID- 16719018 TI - The best and worst aspects of the ICU research coordinator role. AB - BACKGROUND: The Research Coordinator (RC) role is a relative new addition to staffing profiles in Australasian Intensive Care Units (ICUs). The RC plays a pivotal role in conducting ethically and scientifically sound research. There have been anecdotal reports of the RC role in various speciality areas. However, limited research has been undertaken into the role, and only one study has been published from the intensive care setting. AIM: To evaluate which factors the RC found to be the best and worst attributes of the role. METHODS: A cross-sectional web-based cohort study was conducted in 2004 collecting free text information from RCs in ICUs in Australia and New Zealand. RESULTS: Forty-nine participants (71%) completed the study with 273 entries into the two categories of 'best' (60%) and 'worst' (40%) aspects of the role. We identified four thematic clusters in both categories: (1) 'How the job was structured' (2) 'The worth of the job' (3) 'What the work involves' and (4) 'Who I work with'. Both categories received proportionally the same number of responses in each theme with (1) the most frequent and (4) the least number of entries. CONCLUSION: There is much variation between RCs in almost every descriptor of the role. The Australasian Intensive Care RC values autonomy, respect and intellectual stimulation with the scope for extending the role to suit the individual and finds isolation, under-recognition and workload, often not sufficiently compensated, to be the worst aspects of the position. PMID- 16719017 TI - Staff experiences of an early warning indicator for unstable patients in Australia. AB - The objective of this study is to explore nursing and medical staff's perceptions of a clinical marker referral tool implemented to assist in the early identification of unstable patients in the general surgical and medical ward environment. A descriptive, exploratory survey design was undertaken 6 months after the implementation of a clinical marker referral tool. The target population for the survey was all ward nursing, junior medical staff and intensive care unit (ICU) registrars in a metropolitan tertiary referral hospital in Australia. The survey consisted of open-ended and closed-ended questions, as well as statements asking participants to explore their perceptions, attitudes and perceived understanding of the clinical marker referral tool. The surveys were sent to all targeted staff in a personally addressed envelope via the internal mail system. Overall, nursing and medical staff (n = 178) responses were positive to the clinical marker project/tool, offering clear guidelines for staff to respond to the patient's clinical condition and contact the medical staff and the ICU liaison team as appropriate. Furthermore, comments were made in relation to the ICU liaison team acting as a 'support' and 'prompt back-up' for nursing staff when needed. However, ward medical staff had reservations with the clinical markers chosen and with ward nurses being able to contact the ICU registrar after hours. Additionally, the ICU registrars commented on an increase to their workload in having to review unstable patients after hours. These results suggest that the ward nurses required additional support and guidance in caring for the unstable patient in the ward which may improve patient outcomes. Further research investigating the less-favourable responses of the ward medical staff and ICU medical staff is warranted. PMID- 16719020 TI - [Social patrimony and free acces to science]. PMID- 16719019 TI - Triage in the emergency department--a qualitative study of the factors which nurses consider when making decisions. AB - Triage, as a concept, is relatively new in Sweden and means 'sorting'. The triage process was developed to grade patients who needed immediate care. Triage is currently important for the emergency treatment system, and nurses are expected to work with it professionally. The aim of this study is to describe how nurses implement triage when patients arrive at the emergency department of a county hospital, situated in a rural area of Sweden, as well as to highlight the factors considered when prioritizing, in connection with nurses' decision-making. The method used was observations of 19 nurses, with minimal disturbance in their triage work, followed by a short tape-recorded interview, during which the nurses were asked to reflect upon their decision of priorities. Qualitative content analysis of data has been used. The results were divided into two areas, internal factors and external factors. The internal factors reflect the nurse skills and personal capacity. The external factors reflect work environment, including high workload and practical arrangements, and should always be perceived and taken into consideration. Using these factors as a basis, the patients' clinical condition, clinical history, various examinations and tests form an assessment, which subsequently results in a prioritization. PMID- 16719021 TI - [Treatment and temperature series study for use in public health. The case of Castilla-La Mancha, Spain]. AB - BACKGROUND: Numerous articles relate atmospheric variables to health indicators. In large regions, such as Castilla-La Mancha, it may be necessary to divide the region into areas in terms of the atmospheric variables available by selecting a representative weather station for each zone. This article focuses on analyzing the daily temperature data from numerous Castilla La Mancha observatories and reducing the number thereof to a few representative stations for being used in studies relating atmospheric variables to health indicators in this region. METHODS: Castilla-La Mancha weather stations were selected in terms of the number of years available and missing data. After filling in the gaps in the selected series, to detect any possible discontinuities and to homogenize the series, the daily temperature data is used in hierarchical cluster and factorial analyses by principal components. RESULTS: Factorial analyses extract one single factor by using the maximum, mean or minimum temperature series. For the maximum temperatures, this factor explains 93.45% of the variance, with an eigenvalue of 39.249. The "Compuesta" station in Toledo shows correlation coefficients in the principal components matrix of 0.987, 0.991 and 0.981 respectively for the maximum, mean and minimum temperature series. CONCLUSIONS: Castilla-La Mancha is an isoclimatic region in terms of the temperature, the "Compuesta" station in Toledo being selected as the representative station for the region for public health studies. The results afford the possibility of conducting studies broken down into small units such as the provinces, with the stations in the government capitals as a reference. PMID- 16719022 TI - [Cardiovascular risk percentiles avoid overestimation or underestimation of risk calculated by the SCORE risk function]. AB - BACKGROUND: Cardiovascular risk evaluation is of the utmost importance for treatment of patients with cardiovascular risk factors. There are various scales or equations and they can be adapted to a particular population to try to avoid over or under estimation of risk. We propose risk percentiles as a new method of adapting risk scoring systems to our population. METHODS: A cross sectional study of the prevalence of cardiovascular risk factors in the province of Palencia (ERVPA: Cardiovascular Risk Study in Palencia) was conducted. Age, gender, blood pressure, total cholesterol, tobacco consumption and diabetes were evaluated. 514 subjects aged 20 - 79 taken from the general population were studied in 9 health centers from Palencia (Spain). Fatal cardiovascular risk was measured with SCORE project equations. Percentiles were calculated and compared to both equations and a Spearman's correlation coefficient and a kappa agreement coefficient were calculated. RESULTS: The percentages of high risk range from 0% in young subjects to 92% in males over the age of 70 and 67% in females over the age of 70. The concordance with high risk country equations has a kappa coefficient of 0.741. Concordance varies greatly with age and gender. On comparing the percentiles, the concordance between both equations is almost absolute. CONCLUSIONS: Adopting risk percentiles as a method of cardiovascular risk evaluation permits us to adapt any risk equation to a particular area and avoids underestimation or overestimation of risk. Risk percentiles allow us to extrapolate absolute risk for any age. PMID- 16719023 TI - [Health system interventions assessment in Spain: an approach through the analysis of the time trends and the geographical variability of avoidable mortality between 1986 -2001]. AB - BACKGROUND: Many studies have proposed Avoidable Mortality (ME) to monitor the performance of health services although its usefulness is limited by the multiplicity of the avoidable mortality lists being used. Time trends from 1986 2001 and the geographical distribution of avoidable mortality by provinces, are presented for Spain. METHODS: An Avoidable Mortality consensus list is being used. It includes avoidable mortality through the intervention of health services (ISAS in Spanish) and through health policy interventions (IPSI in Spanish). Time trends are analyzed adjusting Poisson or Joinpoint regression models and the annual percentages of change (APC) are estimated. Changes in geographical distribution between the first half of the analysed period and the second are tested by means of standard mortality ratios (SMR) and comparative mortality rates (CMR) for each province. RESULTS: Between 1986 and 2001 avoidable mortality decreased (APC: -1.68; CI: -1.99 and -1.38) slightly more than non-avoidable mortality (APC: -1.28; CI: -1.40 and -1.17). Higher reduction was observed for ISAS mortality (APC: -2.77; CI: -2.89 and -2.65) and an irregular trend for IPSI (between 1986-1990 increase APC: 4.86; CI: 3.32 and 6.41, between 1990-95 stabilization APC: -0.03; CI: -2.32 and 2.31 and finally 1995-2001 decrease APC: 3.57; CI: -4.72 and -2.40). CONCLUSIONS: Avoidable mortality decreased more than non avoidable mortality and important geographical variability can be observed among provinces which should be monitored in order to identify the health services weaknesses. The higher ISAS mortality was observed in southern provinces and the higher IPSI mortality in some areas on the coast. The pattern is somewhat similar for both analyzed periods. PMID- 16719024 TI - [Correspondences from 10th to 9th Revision of the International Classification of Diseases in the causes of death lists of the National Institute of Statistics and the Regional Health Authority of Murcia in Spain]. AB - BACKGROUND: Different countries have conducted comparability studies between Revisions 10 and 9 of the International Classification of Diseases for aggregate lists of causes of death. In Spain, the COMPARA project was aimed at evaluating the impact of the revision change. METHODS: Descriptive cross-sectional epidemiological study of 88,048 deaths recorded in Spain in 1999 with the underlying cause of death doubled coded in ICD-9 and ICD-10. The theoretical correspondences between the ICD on the lists of the National Institute of Statistics and Murcia are established. The comparability rates and their confidence intervals, and the total kappa index were calculated. RESULTS: A decline in infectious diseases (-1.7%) and viral hepatitis, (-12.3%) declined under Tenth revision, while AIDS showed an increase (5.7%). Neoplasms increased a little (0.3%) with the inclusion of the Mielodisplasic Syndrome (55.2%). Diabetes mellitus is increased (2.1%). Mental disorders declined on dementia being shifted to Alzheimer's disease (28.6%). Cardiovascular diseases dropped slightly (-1.4%), without any impact on cerebrovascular diseases, although acute myocardial infarct decreased (-0.6%) while ischemic heart disease increased (0.3%). Pneumonia decreased (-12.5%) and hepatic cirrhosis grows (4.3%). Ill-defined conditions increased due to cardiorespiratory insufficiencies. The external causes show no change without including the accuracy of ICD-9. The National Institute of Statistics 102 groups list obtained a total kappa index of 95.4%, similar to the Murcia variants. CONCLUSIONS: Although ICD-10 has a lesser overall impact, the significant comparability rates of the causes of death groups between the revisions with important absolute differences should be taken into account. PMID- 16719025 TI - [Juan Bautista Juanini: chemical analysis of the air pollution in Madrid (1679)]. PMID- 16719026 TI - A programme of static positional stretches does not reduce hemiplegic shoulder pain or maintain shoulder range of motion--a randomized controlled trial. AB - OBJECTIVE: To evaluate the effectiveness of a programme of static positional stretches and positioning of the stroke-affected shoulder for maintaining shoulder external rotation and decreasing hemiplegic shoulder pain. DESIGN: Randomized controlled trial with pretest and posttest design. SETTING: Inpatient rehabilitation unit. SUBJECTS: Thirty-two participants (17 treatment, 15 comparison) with a first time stroke who were admitted for rehabilitation. INTERVENTIONS: Treatment participants completed a programme of static positional stretches of the stroke-affected shoulder twice daily and positioned the stroke affected upper limb in an armrest support at all other times when seated. MAIN MEASURES: The main outcome measures were pain-free range of motion into external rotation, pain in the stroke-affected shoulder at rest and with movement, motor recovery and functional independence. RESULTS: All participants demonstrated a significant loss of external rotation (P = 0.005) with no significant group differences. All participants demonstrated a significant improvement in motor recovery (P < 0.01) and functional independence (P < 0.01) with no significant group differences. There were no significant effects for pain. The comparison group recorded a decrease in mean pain reported with movement from admission to discharge, and the treatment group recorded an increase. CONCLUSIONS: Participation in the management programme did not result in improved outcomes. The results of this study do not support the application of the programme of static positional stretches to maintain range of motion in the shoulder. The effect of increasing pain for the treatment group requires further investigation. PMID- 16719027 TI - Strapping the hemiplegic shoulder prevents development of pain during rehabilitation: a randomized controlled trial. AB - OBJECTIVE: To determine whether strapping (therapeutic or placebo) the 'at risk' shoulder prevented or delayed development of hemiplegic shoulder pain better than standard care. DESIGN: Randomized controlled trial. SETTING: Three inpatient rehabilitation hospitals, Melbourne, Australia. PATIENTS: Thirty-three patients 'at risk' of developing hemiplegic shoulder pain were randomized, one withdrew. Patients at risk were considered to be those with low or no muscle function around the shoulder (Motor Assessment Scale, upper arm score < 3). INTERVENTIONS: Standard care for control group. Therapeutic strapping or placebo strapping with lightweight adhesive tape. All strapping was maintained over four weeks. MAIN OUTCOME MEASURES: The primary outcome was number of pain-free days measured on Ritchie Articular Index. Secondary outcome measures included range of movement, function (Motor Assessment Scale, upper arm component) and affected arm muscle tone (Modified Ashworth Scale). RESULTS: Only one patient in the therapeutic strapping group developed pain. One-way ANOVA revealed that mean number of pain free days over the 28-day period was significantly different (P = 0.02), with the difference between the therapeutic strapping and the control group. Patients in the therapeutic strapping group had a mean of 26.2 pain-free days, while those in the placebo group and control group had a mean of 19.1 and 15.9 pain-free days respectively. Range of movement and function improved between admission and discharge but were not significantly different between groups. CONCLUSIONS: Therapeutic strapping limited development of hemiplegic shoulder pain during rehabilitation in at risk stroke patients. Placebo strapping has an effect, with a larger study (n = 30 per group) needed to detect whether there are differences between therapeutic and placebo strapping. PMID- 16719028 TI - The effect of a task-oriented intervention on arm function in people with stroke: a randomized controlled trial. AB - OBJECTIVE: To evaluate the efficacy of a task-oriented intervention in enhancing arm function in people with stroke. DESIGN: Two-centre, observer-blinded, stratified, block-randomized controlled trial. SETTING: General community. PATIENTS: Ninety-one individuals within one year of a first or recurrent stroke consented to participate between May 2000 and February 2003. INTERVENTIONS: The experimental intervention involved practice of functional, unilateral and bilateral tasks that were designed to improve gross and fine manual dexterity whereas the control intervention was composed of walking tasks. Members in both groups participated in three sessions a week for six weeks. MAIN OUTCOME MEASURE(S): The primary test of arm function was the Box and Block Test. Secondary tests included the Nine-Hole Peg Test, maximal grip strength, the Test d'Evaluation des Membres superieurs des Personnes Agees (TEMPA) and the Stroke Rehabilitation Assessment of Movement. RESULTS: Results are for the more affected arm. Baseline performance on the Box and Block Test was an average of 26 blocks (standard deviation (SD) = 16) in the experimental group (n = 47) and 26 blocks (SD = 18) in the control group (n = 44). These values represent approximately 40% of age-predicted values. Values for the postintervention evaluation were an average of 28 (SD = 17) and 28 (SD = 19) blocks for the experimental and control group respectively. No meaningful change on other measures of arm function was observed. CONCLUSIONS: A task-oriented intervention did not improve voluntary movement or manual dexterity of the affected arm in people with chronic stroke. PMID- 16719029 TI - Nutritional supplementation and resistance training in nutritionally at risk older adults following lower limb fracture: a randomized controlled trial. AB - OBJECTIVE: To describe the independent and combined effects of oral nutrition supplementation and resistance training on health outcomes in nutritionally at risk older adults following lower limb fracture. DESIGN: Randomized controlled trial with 12-week masked outcome assessment. SETTING: Teaching hospital. PARTICIPANTS: One hundred nutritionally at risk older adults hospitalized following a fall-related lower limb fracture. INTERVENTION: Commenced seven days after injury. Consisted of daily multinutrient energy-dense oral supplement (6.3 kJ/mL) individually prescribed for six weeks (n = 25), tri-weekly resistance training for 12 weeks (n = 25), combined treatment (n = 24) or attention control plus usual care and general nutrition and exercise advice (n = 26). MEASUREMENTS: Weight change, quadriceps strength, gait speed, quality of life and health care utilization at completion of the 12-week intervention. RESULTS: At 12 weeks, all groups lost weight: nutrition -6.2% (-8.4, -4.0); resistance training -6.3% ( 8.3, -4.3); nutrition and resistance training -4.7% (-7.4, -2.0); attention control -5.2% (-9.0, -1.5). Those receiving resistance training alone lost more weight than those receiving the combined treatment (P= 0.029). Significant weight loss was prevented if supplement was consumed for at least 35 days. Groups were no different at 12 weeks for any other outcome. CONCLUSION: Frail, undernourished older adults with a fall-related lower limb fracture experience clinically significant weight loss that is unable to be reversed with oral nutritional supplements. Those receiving a programme of resistance training without concurrent nutrition support are at increased risk of weight loss compared with those who receive a combined nutrition and resistance training intervention. In this high-risk patient group it is possible to prevent further decline in nutritional status using oral nutritional supplements if strategies are implemented to ensure prescription is adequate to meet energy requirements and levels of adherence are high. PMID- 16719030 TI - Immediate effects of an individually designed functional ankle-foot orthosis on stance and gait in hemiparetic patients. AB - OBJECTIVE: To evaluate the immediate effects of individually designed functional in-shoe ankle-foot orthoses (AFO) made of soft and hard cast on balance, standing, and gait parameters in hemiparetic patients. DESIGN: Crossover design with randomized order of the intervention. SETTING: A rehabilitation centre for adults with neurological disorders. SUBJECTS: Twenty-eight patients with hemiparesis due to stroke or traumatic brain injury. MEASURES: Postural sway, standing and gait parameters based on ground reaction forces in two conditions: Patients were randomly assigned to varying sequences of wearing AFO in footwear or wearing footwear alone. RESULTS: AFO significantly improved weight-bearing on the affected leg (affected/ unaffected side symmetry: 2.25 +/- 1.5 with AFO versus 3.4 +/- 2.5 without AFO, P<0.05) and postural sway in stance (12.5 mm +/- 5.2 with AFO versus 15.7 mm +/- 6.7 without AFO, P < 0.05), double stance duration (21.1 +/-14.4% of gait cycle with AFO versus 25.9 +/- 21.6% of gait cycle without AFO, P < 0.05), and symmetry ratios of gait parameters such as stance duration (2.0 +/- 1.5 s with AFO versus 3.3 +/- 3.6 s without AFO, P < 0.05) and deceleration forces (1.6 +/- 0.5 with AFO versus 1.9 +/- 0.6 without AFO, P < 0.05) during gait. No significant differences were observed in all other symmetry ratios of gait parameters. CONCLUSION: An individually designed functional in-shoe AFO can improve stance and gait parameters, even in a single use, in patients with hemiparesis. PMID- 16719031 TI - Validation of self-reported fall events in intervention studies. AB - OBJECTIVE: To determine if self-reported data recalling at least one fall over a six-month recall period could be used as a reliable measure of falls by comparing retrospective self-report with a 'gold standard' method of reporting these events using a prospective calendar-reported method. DESIGN: Prospective study. SETTING: Community-dwelling older people living in New South Wales (NSW) and Queensland in Australia. SUBJECTS: A stratified sample of 264 randomly selected participants from a concurrent randomized controlled trial (RCT) of preventive health assessment for people over the age of 70 years. INTERVENTIONS: Intervention and control group participants in the RCT kept a calendar of fall events for a six month validation substudy. At the end of six months, participants were asked to retrospectively report their falls over the six-month period. MAIN RESULTS: Results indicated that percentage agreement between retrospective self-report and the calendar report of falls was 84%, however, sensitivity was 56% (95% confidence interval (CI) 44.1-67.5). Ten participants (4%) gave false positive self-reports of falls, and 33 (13%) participants gave false negative self-reports of falls. Therefore, retrospective self-reported fall rates were likely to involve under-reporting of falls. Significant differences were found in the sensitivity of retrospectively self-reported falls reported by the intervention group participants (sensitivity = 71%; 95% CI 56.6-85.5) compared with the control group (sensitivity = 40.5%; 95% CI 24.7-56.4, P = 0.008). Validity of self-reported injuries as a result of a fall was less accurate (71% agreement; sensitivity 24% (95% CI 16.0-33.6). CONCLUSIONS: As recall of falls was more accurate in the intervention group than in the control group, it could appear that the intervention had the effect of increasing falls compared to the control group. Using these results, a method was devised to weight self-reported retrospective data to compensate for the poorer recall demonstrated in the control group, so that a more accurate estimate of falls could be derived from the self-reported data. PMID- 16719032 TI - A new clinical test of dynamic standing balance in the frontal plane: the side step test. AB - OBJECTIVE: To confirm retest reliability for the side-step test and to investigate the validity of the side-step test by correlating performance on this test with that on other commonly used measures of balance and mobility. SETTING: Katta Public Hospital, Miyagi, Japan. DESIGN: Cross-sectional study. Intraclass correlation coefficient was examined by calculating the intra-rater reliability of the side-step test. Moreover, the validity of the test was examined by correlating maximum side-step length with gait parameters. SUBJECTS: This study recruited 28 hemiplegic subjects (17 with left-sided hemiplegia and 11 with right sided hemiplegia). MEASURES: We measured balance ability and walking ability as follows: dynamic balance ability was assessed from maximum side-step length from the side-step test, static balance ability from one-footed standing duration, and walking ability from maximum walking speed, stride length and cadence. Balance ability was measured on both the affected and unaffected side. Maximum side-step length was standardized in terms of lower extremity length. RESULTS: Intraclass correlation coefficient of maximum side-step length was 0.97 bilaterally. There was a high linear correlation between maximum side-step length and both maximum walking speed and stride length; Pearson's product-moment correlation coefficient ranged from 0.84 to 0.89. One-footed standing duration was significantly correlated with maximum side-step length, maximum walking speed and stride length, and these relationships were non-linear. CONCLUSION: The reliability and validity of the side-step test was confirmed. PMID- 16719033 TI - Statistical methods for analysing Barthel scores in trials of poststroke interventions: a review and computer simulations. AB - BACKGROUND AND PURPOSE: Arguments persist as to whether parametric or non parametric methods should be used to analyse ordinal data in trials. This paper aims to assess methods used for presenting and analysing an ordinal scale, the Barthel Index, in trials of poststroke interventions. METHODS: All randomized controlled trials (RCTs) of poststroke interventions published from 1995 to 2004 in two journals (Stroke and Clinical Rehabilitation) were scrutinized for methods used to present and analyse Barthel scores. Computer simulations were used to compare the type I errors and the statistical power of different statistical methods under a range of assumed circumstances. RESULTS: One hundred and fifty six RCTs were identified within the two journals. The central tendency of Barthel scores was measured by the median in 47 trials and by the mean in 35 trials. Non parametric analyses of Barthel scores were conducted in 47 trials and parametric methods used in 18 trials. The results of computer simulations demonstrate that the t-test has a similar type I error rate and statistical power when compared with the rank sum test. However, when a zero final Barthel score is assigned to patients who have died, the statistical power of the t-test is much reduced. The possible maximal statistical power of dichotomization and ordinal regression is usually much lower than that of the rank sum test. CONCLUSIONS: To facilitate comparison and meta-analysis, we recommend that mean values (with standard deviations or standard errors) of Barthel scores should be routinely reported in trials of poststroke interventions. The rank sum test appears the most powerful inferential technique for detecting differences in Barthel scores. PMID- 16719034 TI - Determinants of physical health in frail older people: the importance of self efficacy. AB - OBJECTIVE: There is continued uncertainty regarding the strength of association between performance-based and self-report measures of physical functioning, and of their relationship to self-efficacy and health-related quality of life (HRQoL). This study assessed the inter-relationships between such measures, and the predictors of 'physical' aspects of HRQoL in frail older patients. DESIGN: We used statistical models to determine the predictors of 'physical' HRQoL, according to the physical component summary score and the physical functioning domain of the 36-item short form (SF-36) questionnaire. SETTING: Patients were recruited from hospitals in Australia and New Zealand and followed up in their homes. SUBJECTS: Two hundred and forty-three frail older patients. MAIN MEASURES: Physical functioning was assessed using three performance-based measures (Timed Up and Go Test, gait speed and the Berg Balance Scale) and five self-report measures, including the modified falls self-efficacy scale, at three and six months after registration. RESULTS: A moderate association (r = 0.48-0.55) was found between each of the performance-based and self-report measures, including the SF-36 physical component summary score. Multiple linear regression analyses showed that the performance-based measures and falls self-efficacy predicted 33% of the SF-36 physical component summary score. Falls self-efficacy was the single highest predictor of both the SF-36 physical component summary score and SF-36 physical functioning domain. A curvilinear relationship was found between the SF 36 physical functioning domain and two variables: falls self-efficacy and the Berg Balance Scale. CONCLUSIONS: Although performance-based and self-report measures provide complementary but distinct measures of physical function, psychosocial factors such as self-efficacy have a strong influence on the HRQoL of frail older people. PMID- 16719035 TI - Appreciating the best. PMID- 16719036 TI - Toys--friend or foe? A study of infection risk in a paediatric intensive care unit. AB - Toys are an established part of the hospital experience for the child and family. They are seen as a source of comfort and security and form part of the child friendly environment. However, they can also act as a source of healthcare associated infection which can be harmful to children, especially those who are in intensive care environments. This small-scale study was conducted in a paediatric intensive care unit at a large teaching hospital and involved swabbing those toys that had been brought in by families and those that were provided by the hospital. Findings show that 85 per cent of the toys harboured viable bacteria, which could be damaging to the child's health. Recommendations for practice are identified to ensure that toys remain a component of the child's hospitalisation yet are safe in relation to the transmission of infections not just in PICU but in all paediatric settings. PMID- 16719037 TI - The psychosocial impact on parents of tube feeding their child. AB - A review of 13 papers investigating parents' experience of long-term tube feeding in disabled children and young people identified a significant impact on parents and families related to oral feeding, decision making and tube feeding itself. Mixed messages and pressure from health professionals and relatives made decision making about tube feeding more difficult for parents. Making the decision to tube feed or proceed to gastrostomy was described in terms of 'giving in'. Parents expressed a need for consistent, accurate information. Once tube feeding was established there is a positive impact on the lives of the child and family - although some parents reported reduced support and continued feelings of inadequacy. The significance parents attach to oral feeding and their information and respite care needs when tube feeding must be recognised and further explored. PMID- 16719038 TI - Levels of continence in children with cerebral palsy. AB - Little is known about the levels of continence in children with cerebral palsy and what factors affect this. This study was conducted to determine levels of urine and stool continence in children with cerebral palsy in relation to their learning disability (LD) and mobility. The aim was to enable us to predict level of continence that can be achieved for these children with a given level of learning disability and mobility. Data were obtained from medical notes and by telephone interview on mobility, degree of learning disability and urine and stool continence of 55 children in Southern Derbyshire. Most of the children who have nil to moderate disabilities in learning and mobility were able to achieve a decent level of continence around a median age of three years. Therefore, their toilet training should be started at the usual age with ongoing support from parents/carers. In children with severe learning disability and severe immobility, the probability of continence is very slim after the age of eight years. After this age it may be better to divert resources to other aspects of care, for example nutrition and physiotherapy, to improve their quality of life. PMID- 16719039 TI - 'Being on the other side': a mother's experience of intensive care. PMID- 16719040 TI - Experiences of mothers of children with Down syndrome. AB - This qualitative study explored the experiences and lifestyles of families in Turkey with children with Down syndrome, including the impact on family members. Twelve mothers with a Down syndrome child (three from each of the age groups 1-3 years, 4-6 years, 7-12 years and 13-18 years of age) participated in the study. The data were collected during in-depth interviews and were evaluated using qualitative data analysis methods. Families were affected socially, physically, economically and emotionally by having a child with Down syndrome. PMID- 16719041 TI - Preparing children and young people for stoma surgery. AB - There is little published literature regarding children and young people with stomas and their preoperative care. Research in adults reveals negative psychosocial consequences, but the perceptions of children and young people may be different, depending on the reason for the stoma formation and the degree of involvement in decisions. Post-operative problems and later negativity can be minimised with informed decision making and partnership from the outset between the healthcare team, parents and child. Preparation approaches known to be successful for other forms of surgery can be adapted to prepare the child/young person for stoma surgery. PMID- 16719042 TI - An evaluation of two subcutaneous infusion devices in children receiving palliative care. AB - Children and young people with advanced cancer frequently have a central venous access device (central line) in place which can be used for intravenous medication for symptom management. Those that do not may need a subcutaneous device for medication infusion. Two subcutaneous devices were evaluated in four children requiring infusion of opioids, anti-emetics, anti-muscarinic agents and benzodiazepines during their palliative care. The length of time the device was in situ and local skin reactions were monitored and the views of children, parents and nurses obtained. Out of 32 evaluations, 28 (88 per cent) indicated that the Thalaset s/c device was preferred to the winged device in terms of child comfort and nurse satisfaction. Although limited by the number of children in the evaluation, the results support the use of the Thalaset s/c device as part of symptom management for children and young people who do not have central lines. PMID- 16719043 TI - Palliative care for children: a public health initiative. AB - This article draws parallels between the work of a children's palliative care team and the health promoting/ prevention focus of public health. From the primary prevention role in supporting decisions about screening for genetic disorders to advocacy for adequate respite, the palliative care team makes a significant but largely unrecognised contribution to community health. Reframing the palliative care service in this way may help organisations responsible for commissioning and delivering services to more clearly see the essential benefits to the wider community of such services and to locate them within their planning priorities. PMID- 16719044 TI - Pig-to-nonhuman primate islet xenotransplantation: a review of current problems. AB - Islet allotransplantation has been shown to have potential as a treatment for type 1 diabetic patients. Xenotransplantation, using the pig as a donor, offers the possibility of an unlimited number of islets. This comprehensive review focuses on experience obtained in pig-to-nonhuman primate models, particularly with regard to the different types of islets (fetal, neonatal, adult) and isolation procedures used, and the methods to determine islet viability. The advantages and disadvantages of the methods to induce diabetes (pancreatectomy, streptozotocin) are discussed. Experience in pig-to-nonhuman primate islet transplantation studies is reviewed, including discussion of the possible mechanisms of rejection and the immunosuppressive regimens used. The research carried out to date has led to workable animal models to study islet xenotransplantation, but several questions regarding methodology remain unanswered, and details of these practicalities require to be adequately addressed. The encouraging porcine islet survival reported recently provides an indicator for future immunosuppressive regimens. PMID- 16719045 TI - Coinhibitory T-cell signaling in islet allograft rejection and tolerance. AB - Autoaggressive T cells directed against insulin secreting pancreatic beta-cells mediate the development of type 1 diabetes. Islet transplantation offers superior glycemic control over exogenous insulin, but chronic immunosuppression limits its broad application. Pathogenic T cells are also important in allograft rejection. Inducing and maintaining antigen-specific peripheral T-cell tolerance toward beta cells is an attractive strategy to prevent autoimmune disease, and to facilitate treatment of diabetes with islet allografts without long-term immunosuppression. Recent efforts have focused on blocking costimulatory T-cell signals for tolerance induction. Although costimulatory blockade can prolong graft survival, true immunological tolerance remains elusive. Costimulatory signals may even be required for the maintenance of peripheral tolerance. The discovery of novel coinhibitory T-cell pathways, including CTLA-4, PD-1, and BTLA, offers an alternative approach. Stimulating negative T cell cosignals alone or in combination may help induce tolerance. The focus of this review is to summarize the strategies directed at turning off the immune response by exploiting these negative cosignaling pathways in tolerance induction in islet transplantation. Activating several coinhibitory pathways together may be synergistic in preventing pathogenic T-cell responses. Tolerance induction will likely rely on understanding the balance of positive and negative signals affecting the state of T-cell activation. PMID- 16719046 TI - Viability and functionality of bovine chromaffin cells encapsulated into alginate PLL microcapsules with a liquefied inner core. AB - Implantation of adrenal medullary bovine chromaffin cells (BCC), which synthesize and secrete a combination of pain-reducing neuroactive compounds including catecholamines and opioid peptides, has been proposed for the treatment of intractable cancer pain. Macro- or microencapsulation of such cells within semipermeable membranes is expected to protect the transplant from the host's immune system. In the present study, we report the viability and functionality of BCC encapsulated into microcapsules of alginate-poly-L-lysine (PLL) with a liquefied inner core. The experiment was carried out during 44 days. Empty microcapsules were characterized in terms of morphology, permeability, and mechanical resistance. At the same time, the viability and functionality of both encapsulated and nonencapsulated BCC were evaluated in vitro. We obtained viable BCC with excellent functionality: immunocytochemical analysis revealed robust survival of chromaffin cells 30 days after isolation and microencapsulation. HPLC assay showed that encapsulated BCC released catecholamines basally during the time course study. Taken together, these results demonstrate that viable BCC can be successfully encapsulated into alginate-PLL microcapsules with a liquefied inner core. PMID- 16719047 TI - One-step induction of neurons from mouse embryonic stem cells in serum-free media containing vitamin B12 and heparin. AB - We present a simple method for neural cell fate specification directly from mouse embryonic stem cells (ES cells) in serum-free conditions in the absence of embryoid body formation. Dissociated ES cells were cultured in serum-free media supplemented with vitamin B12 and heparin, but without any expensive cytokines. After 14 days in culture, beta-tubulin type III (TuJ1) and tyrosine hydroxylase (TH)-positive colonies were detected by immunocytochemical examinations. In addition, specific gene analyses by RT-PCR demonstrated expression of an early central nerve system, mature neuron, and midbrain dopaminergic neuron-specific molecules (i.e., nestin, middle molecular mass neurofilament protein, Nurr1, and TH, respectively). Dopamine was also detected in the culture media by reverse phase HPLC analysis. These facts indicate that addition of vitamin B12/heparin to serum-free culture media induced neurons from ES cells, which included cells that released dopamine. Other supplements, such as putrescine, biotin, and Fe2+, could not induce neurons from ES cells by themselves, but produced synergistic effects with vitamin B12/heparin. The rate of TuJ1+/TH+ colony formation was increased threefold and the amounts of dopamine released increased 1.5-fold by the addition of a mixture of putrescine, biotin, and Fe2+ to vitamin B12/heparin culture media. Our method is a simple tool to differentiate ES cells to dopaminergic neurons for the preparation of dopamine-releasing cells for the cell transplantation therapy of Parkinson's disease. In addition, this method can facilitate the discovery of soluble factors and genes that can aid in the induction of the ES cell to its neural fate. PMID- 16719048 TI - Engrafted neural progenitor cells express a tissue-restricted reporter gene associated with differentiated retinal photoreceptor cells. AB - Neural progenitor cells (NPCs) have shown ability to repair injured CNS, and might provide precursors to retinal neurons. NPCs were isolated from the brains of 14 day murine embryos of transgenic mice that express beta-galactosidase (beta gal) on the arrestin promoter, which specifically directs expression to retinal photoreceptor cells. NPCs were transferred to adult, syngeneic mice via inoculation into the anterior chamber of the eye, the peritoneal cavity, or the brain. At 14 weeks postgrafting, tissues were collected and examined to determine if differentiated NPC progeny were present in retina based on histochemical detection of beta-gal. Four of six anterior chamber-inoculated recipients showed Bluo-gal-stained cells in retina, indicating the presence of transferred NPCs or their progeny. Because the progenitor cells do not express beta-gal, positive staining indicates differentiation leading to activation of the arrestin promoter. Two recipients inoculated by the intraperitoneal route also exhibited Bluo-gal staining in retina. The NPCs did not express beta-gal if inoculated into brain, but survived and dispersed. Most recipients, regardless of inoculation route, were PCR positive for beta-gal DNA in extraocular tissues, but no Bluo-gal staining was found outside of the retina. Injury to the retina promoted, but was not required, for progenitor cell engraftment. beta-Gal-positive cells were concentrated in the outer layers of the retina. In summary, a reporter gene specifically expressed in differentiated retinal photoreceptor cells due to the activity of the arrestin promoter was expressed in recipient mouse retina following transfer of NPCs prepared from the beta-gal transgenic mice. The presence of beta-gal DNA, but not Bluo-gal staining, in spleen and other tissues revealed that the cells also migrated elsewhere and took up residence in other organs, but did not undergo differentiation that led to beta-gal expression. PMID- 16719049 TI - Subnormothermic preservation maintains viability and function in a porcine hepatocyte culture model simulating bioreactor transport. AB - Bioartificial liver (BAL) systems have been developed to bridge patients with acute liver failure (ALF) to liver transplantation or liver regeneration. Clinical application of BAL systems is dependent on the supportive quality of cells used and direct availability of the whole system. Reliable transport of BAL systems from the laboratory to remote treatment centers is therefore inevitable. Subsequently, preservation conditions play a crucial role during transport of a BAL, with temperature being one of the most determining factors. In this study, we assessed the effect of subnormothermic preservation on freshly isolated porcine hepatocytes cultured in monolayer under oxygenation. Additionally, the effect of the University of Wisconsin (UW) preservation solution was compared with Williams' E (WE) culture medium at 4 degrees C. The control group was cultured for 3 days at 37 degrees C, whereas the transport groups were cultured at 4 degrees C, 15 degrees C, 21 degrees C, or 28 degrees C for 24 h at day 2. All groups were tested each day for cell damage and hepatic functions. Subnormothermic culture (i.e., 15 degrees C to 28 degrees C) for a period of 24 h did not reduce any hepatic function and did not increase cellular damage. In contrast, culture of hepatocytes in WE medium and preservation in UW solution at 4 degrees C significantly reduced hepatic function. In conclusion, freshly isolated porcine hepatocytes can be preserved for 24 h at subnormothermic temperatures as low as 15 degrees C. Future research will focus on the implementation of the AMC-BAL in an oxygenated culture medium perfusion system for transport between the laboratory and the hospital. PMID- 16719051 TI - Comparison of cooling systems during islet purification. AB - Islet isolation is a complex procedure that includes digestion and purification of pancreatic tissue. As we move towards clinical regulatory control and standardization, understanding of the detailed stages of the procedure have become increasingly important. Purification on a COBE 2991 density gradient allows human islets to be separated from a large volume of acinar tissue. Cooling the gradient and tissue is thought to be important to reduce metabolic activity but cooling systems for the gradient are expensive, with limited availability. In this study, the efficiency of cooling methods for the COBE 2991 cell separator has been investigated. The two cooling systems were: a) COBE 2991 modified internally to allow coolant (polyethylene glycol) from a chiller to circulate either side of the spindle and around the bowl (original system), and b) an air cooled system using an air conditioner to blow cold air into the bowl from above (air cooler system). Cooling required 20 min for the original system and temperature was stabilized within 4-7 degrees C. The air system cooled rapidly but was not stable. There was an increase in the temperature of the medium with using both systems during centrifugation because of heat generated by the COBE machine; however, the temperature of the medium after centrifugation with the air system was significantly higher than that with the original system (13.3 +/- 0.2 degrees C vs. 8.7 +/- 0.7 degrees C, p < 0.05). The original cooler system was found to be more efficient at reducing heat generated by the COBE machine than the air system. Further investigation of the importance of the recorded temperatures is required. PMID- 16719050 TI - Quantitative assay for quality assurance of human cells for clinical transplantation. AB - Transplantation of human cells after isolation and culture has become an important alternative for treatment of acute or chronic skin wounds. To increase the efficacy and reduce cost for transplantation of skin cells, more efficient and accurate techniques for evaluation of cell proliferation are needed. Hemocytometer counts provide a valid assessment of cell proliferation and viability, but they are very labor intensive and require removal of the cells from their substrate. In this study, hemocytometer counts were compared with a fluorometric assay (n = 21 per condition) that uses the commercially available reagent alamarBlue, which is reduced to a fluorescent substrate by cellular dehydrogenases. Human epidermal keratinocytes were inoculated at 200, 600, 2000, and 6000 cells/cm2 incubated for 6 days in modified MCDB 153 medium. Alamar Blue was incubated with cells for 2 h at 37 degrees C, and fluorescence was measured with a microplate reader at 590 nm. Hemocytometer counts (x10(-4)) from the respective cell inoculation densities were 0.30 +/- 0.04, 1.07 +/- 0.10, 6.37 +/- 0.62, and 16.99 +/- 0.96. Fluorescence values (x10(-3)) for the respective inoculation densities were 0.14 +/- 0.01, 0.34 +/- 0.02, 1.20 +/- 0.09, and 1.79 +/- 0.12. Regression analysis showed a statistical significant (p < 0.0001) correlation (r2 = 0.87) between cell counts and optical density from the alamarBlue assay. These data demonstrate that alamarBlue provides a valid substitute for cell counts to assess cell proliferation before clinical transplantation of engineered skin. AlamarBlue also allows repeated, nondamaging assessment of living cells over time. These advantages are expected to increase the validity and reliability of quality assurance standards for transplanted skin cells, and to increase the efficacy of healing of cutaneous wounds. PMID- 16719052 TI - Estimation of pancreas weight from donor variables. AB - Previous studies have identified several donor factors affecting the outcome of islet isolation. Pancreas weight has not been considered as a donor selection criterion, because a value cannot be obtained prior to organ procurement. However, a larger pancreas will likely contain a higher number of islets. Therefore, the prediction of pancreas weight would be helpful in donor selection, benefiting cost and efficiency of the islet isolation laboratory. The purpose of this study was to investigate normal pancreas weight in cadaveric donors and identify pancreas weight predictors from demographic data of cadaveric organ donors. We retrospectively analyzed data on pancreas weight from 354 cadaveric donors with respect to gender, age, body weight, body height, body mass index (BMI), and body surface area (BSA). In men, pancreas weight correlated more closely with body weight than with age, height, or BMI. BSA was as strong a correlate of pancreas weight as body weight. In women, pancreas weight had a similar pattern of relationships, with generally lower correlation coefficients. On the basis of the observation of gender-specific pancreas weight difference in elderly donors, stepwise multiple linear regression analyses were conducted separately for younger (< or =40 years) and elderly (> or =41 years) donors. In younger donors, body weight and age were the major predictors of pancreas weight [pancreas weight (g) = 4.355 + 0.742 x body weight (kg) + 0.837 x age (years) (R2 = 0.564, p < 0.001)]. In contrast, pancreas weight of elderly donors was best predicted by BSA and gender [pancreas weight (g) = -17.624 + 60.036 x BSA (m2) - 7.152 x gender (R2 = 0.372, p < 0.001; "gender": 1 = female, 0 = male)]. Pancreas weight was found to be positively associated with pre- and postpurification islet yields. These formulae should contribute to the estimation of pancreas weight, and thus improve donor selection for islet isolation and transplantation. PMID- 16719053 TI - Ameliorating injury during preservation and isolation of human islets using the two-layer method with perfluorocarbon and UW solution. AB - This study assessed the effects of a two-layer method (TLM), using perfluorocarbon and UW solution, on the quality of human pancreata following storage and islet yield/function after isolation. In part A, TLM was applied immediately after procurement and the energetic profile was compared to a group treated with UW solution only (control) throughout 24-h storage. In part B, cadaveric human pancreata were procured and subjected to a TLM after cold storage in UW solution (TLM group) or UW solution (control group). Energetics, lipid peroxidation, and islet recovery/function were assessed after preservation at 4 degrees C. In part A, after 9-h storage, the energetic profile (ATP, ATP/ADP, energy charge) for the TLM group was superior to controls. In part B, TLM treatment resulted in consistently greater ATP, ATP/ADP, and energy charge values than with storage in UW solution alone (p < 0.05). UW treatment resulted in 40% greater peroxidative damage than in the TLM group (p < 0.05). Islet recovery and functional viability were 30-40% higher following TLM treatment (p < 0.05). These data support the hypothesis that islet viability and yields can be significantly improved using a brief period of TLM treatment following conventional UW storage; reduced energetic and oxidative stress are implicated as potential mechanisms. PMID- 16719054 TI - Functional MR microimaging of pancreatic beta-cell activation. AB - The increasing incidence of diabetes and the need to further understand its cellular basis has resulted in the development of new diagnostic and therapeutic techniques. Nonetheless, the quest to noninvasively ascertain beta-cell mass and function has not been achieved. Manganese (Mn)-enhanced MRI is presented here as a tool to image beta-cell functionality in cell culture and isolated islets. Similar to calcium, extracellular Mn was taken up by glucose-activated beta-cells resulting in 200% increase in MRI contrast enhancement, versus nonactivated cells. Similarly, glucose-activated islets showed an increase in MRI contrast up to 45%. Although glucose-stimulated Ca influx was depressed in the presence of 100 microM Mn, no significant effect was seen at lower Mn concentrations. Moreover, islets exposed to Mn showed normal glucose sensitivity and insulin secretion. These results demonstrate a link between image contrast enhancement and beta-cell activation in vitro, and provide the basis for future noninvasive in vivo imaging of islet functionality and beta-cell mass. PMID- 16719055 TI - Size-dependent revascularization of transplanted pancreatic islets. AB - For their survival and optimal function, pancreatic islets depend posttransplantation on a rapid and adequate revascularization. Native islets display a marked size-dependent heterogeneity in both angioarchitecture and degree of blood perfusion. This study evaluated whether there also are differences in the degree of revascularization of islets of different size when transplanted. Mouse pancreatic islets were isolated by collagenase digestion, and cultured in vitro for 4-7 days before transplantation. Groups of 200 islets with a diameter either exceeding or being below 100 microm were implanted beneath the left renal capsule of syngeneic C57 BL/6 mice. One month posttransplantation, graft-bearing kidneys were removed. Histological specimens were prepared and stained for endothelium with the lectin Bandeiraea simplicifolia. Pancreata from nontransplanted control animals were prepared similarly. The vascular density in transplanted islets was markedly lower than in native islets. However, islet transplants composed of small islets (<100 microm in diameter) had a vascular density in the endocrine tissue twice that in transplants of larger islets (>100 microm). The connective tissue stroma surrounding smaller islets was also more revascularized than in corresponding grafts with large islets. The vascular density in the connective tissue stroma surrounding the individual islets in the grafts was markedly higher than in the endocrine parts per se. These combined observations indicate that smaller islets have a higher capacity to stimulate regrowth of blood vessels following transplantation. Further studies on islet differences with regard to revascularization capacity may teach us strategies for treatment of transplanted islets to improve their revascularization. PMID- 16719056 TI - Timing of cord blood treatment after experimental stroke determines therapeutic efficacy. AB - Embolic stroke is thought to cause irreparable damage in the brain immediately adjacent to the region of reduced blood perfusion. Therefore, much of the current research focuses on treatments such as anti-inflammatory, neuroprotective, and cell replacement strategies to minimize behavioral and physiological consequences. In the present study, intravenous delivery of human umbilical cord blood cells (HUCBC) 48 h after a middle cerebral artery occlusion (MCAo) in a rat resulted in both behavioral and physiological recovery. Nissl and TUNEL staining demonstrated that many of the neurons in the core were rescued, indicating that while both necrotic and apoptotic cell death occur in ischemia, it is clear that apoptosis plays a larger role than first anticipated. Further, immunohistochemical and histochemical analysis showed a diminished and/or lack of granulocyte and monocyte infiltration and astrocytic and microglial activation in the parenchyma in animals treated with HUCBC 48 h poststroke. Successful treatment at this time point should offer encouragement to clinicians that a therapy with a broader window of efficacy may soon be available to treat stroke. PMID- 16719057 TI - Genetically engineered human mesenchymal stem cells produce met-enkephalin at augmented higher levels in vitro. AB - We have reported that transplantation of adrenal medullary chromaffin cells that release endogenous opioid peptides into pain modulatory regions in the CNS produce significant antinociceptive effects in patients with terminal cancer pain. However, the usefulness of this procedure is minimal because the availability of human adrenal tissue is very limited. Alternative xenogeneic materials, such as porcine and bovine adrenal chromaffin cells present problems of immune rejection and possible pathogenic contamination. In an attempt to develop opioid peptide-producing cells of autologous origin, we have transfected human mesenchymal stem cells (hMeSCs) with a mammalian expression vector containing a fusion gene of green fluorescent protein (GFP) and human preproenkephalin (hPPE), a precursor protein for enkephalin opioid peptides. Enkephalins are major neurotransmitters that play an important role in analgesia by activating peripheral opioid receptors. Following the establishment of stable transfection of hMeSCs, the expressions of hPPE and GFP were confirmed and the production of methionine enkephalin (Met-enkephalin) was significantly increased compared to control naive hMeSCs (p < 0.05). Our in vitro data demonstrated that genetically engineered hMeSCs with transfected hPPE gene can constitutively produce opioid peptide Met-enkephalin at an augmented high level. hMeSCs are relatively easy to isolate from a patient's bone marrow aspirates and expand in culture by repeated passages. Autologous hMeSCs would not require immunosuppression when transplanted back into the same patient. Through targeted gene manipulation such as hPPE gene transfection, this may offer a virtually unlimited safe cell supply for the treatment of opioid-sensitive pain in humans. PMID- 16719058 TI - Behavioral and histological characterization of intrahippocampal grafts of human bone marrow-derived multipotent progenitor cells in neonatal rats with hypoxic ischemic injury. AB - Children born with hypoxic-ischemic (HI) brain injury account for a significant number of live births wherein no clinical treatment is available. Limited clinical trials of stem cell therapy have been initiated in a number-of neurological disorders, but the preclinical evidence of a cell-based therapy for neonatal HI injury remains in its infancy. One major postulated mechanism underlying therapeutic benefits of stem cell therapy involves stimulation of endogenous neurogenesis via transplantation of exogenous stem cells. To this end, transplantation has targeted neurogenic sites, such as the hippocampus, for brain protection and repair. The hippocampus has been shown to secrete growth factors, especially during the postnatal period, suggesting that this brain region presents as highly conducive microenvironment for cell survival. Based on its neurogenic and neurotrophic factor-secreting features, the hippocampus stands as an appealing target for stem cell therapy. Here, we investigated the efficacy of intrahippocampal transplantation of multipotent progenitor cells (MPCs), which are pluripotent progenitor cells with the ability to differentiate into a neuronal lineage. Seven-day-old Sprague-Dawley rats were initially subjected to unilateral HI injury, which involved permanent ligation of the right common carotid artery and subsequent exposure to hypoxic environment. At day 7 after HI injury, animals received stereotaxic hippocampal injections of vehicle or cryopre served MPCs (thawed just prior to transplantation) derived either from Sprague Dawley rats (syngeneic) or Fisher rats (allogeneic). All animals were treated with daily immunosuppression throughout the survival period. Behavioral tests were conducted on posttransplantation days 7 and 14 using the elevated body swing test and the rotarod to reveal general and coordinated motor functions. MPC transplanted animals exhibited reduced motor asymmetry and longer time spent on the rotarod than those that received the vehicle infusion. Both syngeneic and allogeneic MPC transplanted injured animals did not significantly differ in their behavioral improvements at both test periods. Immunohistochemical evaluations of graft survival after behavioral testing at day 14 posttransplantation revealed that syngeneic and allogeneic transplanted MPCs survived in the hippocampal region. These results demonstrate for the first time that transplantation of MPCs ameliorated motor deficits associated with HI injury. In view of comparable behavioral recovery produced by syngeneic and allogeneic MPC grafts, allogeneic transplantation poses as a feasible and efficacious cell replacement strategy with direct clinical application. An equally major finding is the observation lending support to the hippocampus as an excellent target brain region for stem cell therapy in treating HI injury. PMID- 16719059 TI - Progressive dopamine neuron loss in Parkinson's disease: the multiple hit hypothesis. AB - Animal models have been an essential tool for researchers and clinicians in their efforts to study and treat Parkinson's disease (PD). Thus, the various ways 6 hydroxydopamine is employed, the use of MPTP in rodents and nonhuman primates, the prenatal exposure to bacterial endotoxin, the postnatal exposure to environmental toxins such as paraquat and rotenone, the assessment of dopamine (DA) neurons in genetic knockout mouse, and even the behavioral analysis of fruit flies and worms have added significantly to our knowledge base of PD--or have they? Are these animal models manifesting a true model of PD? Have the 7786 published studies (to date) on PD with animal models led to a clearer understanding of its etiology, treatment, or progression? In this review we critically assess this question. We begin with a succinct history of the major contributions, which have led to the current animal models of PD. We then evaluate the primary issue of the progressive loss of DA neurons, which, except for a few studies, has not been addressed in animal models of PD, even though this is the major pathological characteristic of the disease. Lastly, we discuss the possibility that more than one risk factor for PD may be necessary to develop an animal model that shows synergy--the progressive loss of DA neurons. Thus, the multiple hit hypothesis of PD-that is, the effect of more then one risk factor may be the start of new era in animal models of PD that is one step closer to mimicking the pathology of PD in humans. PMID- 16719060 TI - Neural repair strategies for Parkinson's disease: insights from primate models. AB - Nonhuman primate models of Parkinson's disease (PD) have been invaluable to our understanding of the human disease and in the advancement of novel therapies for its treatment. In this review, we attempt to give a brief overview of the animal models of PD currently used, with a more comprehensive focus on the advantages and disadvantages presented by their use in the nonhuman primate. In particular, discussion addresses the 6-hydroxydopamine (6-OHDA), 1-methyl-1,2,3,6 tetrahydopyridine (MPTP), rotenone, paraquat, and maneb parkinsonian models. Additionally, the role of primate PD models in the development of novel therapies, such as trophic factor delivery, grafting, and deep brain stimulation, are described. Finally, the contribution of primate PD models to our understanding of the etiology and pathology of human PD is discussed. PMID- 16719061 TI - CNS gene therapy and a nexus of complexity: systems and biology at a crossroads. AB - Gene therapy is a potentially promising new treatment for neurodegenerative disorders such as Alzheimer's disease (AD), which has been difficult to treat with conventional therapeutics. Viral vector-mediated somatic gene therapy is a rapidly developing methodology for providing never before achieved capability to deliver specific genes to the CNS in a highly localized and controlled manner. With the advent and refinements of this technology one focus is directed to which genes are the most appropriate to select for specific disease indications. Nerve growth factor (NGF), a potent survival factor for critical cell populations that degenerate in AD, has been chosen already for clinical gene therapy trials in human AD patients. Much knowledge about the pathophysiological underpinnings of AD is still lacking to make clear which patients may benefit from a gene therapy approach. Moreover, a detailed understanding of sustained NGF action in the normal and diseased CNS needs to be resolved before conclusions can be drawn regarding the utility of NGF gene therapy. Systematic efforts to acquire this new knowledge should compel clinically and biologically sophisticated efforts to advance gene therapy for neurodegenerative diseases. PMID- 16719062 TI - [Complex treatment of patients with postburn esophageal stricture]. AB - Results of treatment of 136 patients with postburn esophageal stricture were studied. In 1998-2001 yrs. there were treated 77 patients using bougienage and operative method, in 2002-2005 yrs bougienage, balloon dilation, prolonged balloon dilation, insertion of endoprosthesis, operative treatment were performed in 59 patients. Simultaneous application of methods of treatment have guaranteed the excellent and good results obtaining in up to 73% of patients. PMID- 16719063 TI - [Diagnosis and treatment of Zollinger-Ellison syndrome]. AB - In 1970-2005 yrs. 65 patients with Zollinger-Ellison syndrome were observed and operated on in the clinic. The decisive meaning in diagnosis owes radioimmunological determination of gastrin level in the blood and its changes while conduction of tests with calcium and secretin. Surgical tactics was determined by localization, number and character of gastrinomas. During the first period of work gastrectomy with complete excision of gastrinproducing tumor constituted the operation of choice. Implementation of intraoperative method of ultrasonography have permitted to excise the benign gastrinoma when her localization was favourable with preservation of stomach. Minimal life span after gastrectomy, performed for nonresectable malignant gastrinoma, have constituted 9 years. PMID- 16719064 TI - [Application of cyprolet in abdominal surgery in a complex antibiotic prophylaxis]. AB - In 90 patients with strangulated inguinal hernia without signs of intestinal necrosis antibioticoprophylaxis was conducted, which included antibiotic Cyprolet (cyprofloxacine) manufactured by "Dr. Reddy's" company, what have promoted the risk reduction of the purulent-septic complications occurrence. The antibacterial preparation choice was preceded by investigation of the exudates microbic spectrum, what have permitted the pathogenetic substantiation of expedient specifical application Cyprolet for antibioticoprophylaxis in conditions of emergent surgery. PMID- 16719065 TI - [Combined surgical treatment of duodenal ulcer disease and reflux-esophagitis]. AB - Surgical treatment was conducted in 328 patients with duodenal ulcer disease and reflux-esophagitis. In all the patients selective proximal vagotomy and correcting operation on cardia was performed. All the patients survived after the operation. There were examined 250 (76.2%) patients in terms up to 20 years, good and satisfactory results were noted in 98.4%, poor--in 1.6%. PMID- 16719066 TI - [Surgical treatment of perforative gastroduodenal ulcer in patients living in a catastrophic flood zone]. AB - In a period of catastrophic flood in 1998 and 2001 yrs there was noted the enhancement of general quantity of patients with gastroduodenal ulcer disease as well as with complicated course, including gastroduodenal ulcer perforation. In the period of flood in Zakarpattya in 2001 the quantity of patients, who were admitted to hospital, to Tyachiv and Vinogradiv central district hospitals in particular, too late, was enhanced as twice. In conditions of extreme situation it is expedient to conduct prophylactic treatment of patients, who are staying under the dispensary observation for gastroduodenal ulcer disease. Improvement of the surgical treatment results in patients with gastroduodenal ulcer disease complications in the disaster zone may be achieved due to accurate organization and coordination on all the stages of medical care to population, application of modern endoscopic technologies. PMID- 16719067 TI - [An early ileus after operations performed on the intestine]. AB - The results of treatment of 87 patients, in whom an early ileus have occurred after performance of operations on the intestine, were analyzed. The main unresolved problems are a timely diagnosis and adequate tactics of treatment. There was proposed the method of their own of the short intestinal loop syndrome prophylaxis after performance of the small intestine extensive resection. Complications have had occurred in 12.6% of patients, postoperative mortality was 3.4%. Elaboration of effective methods of the adhesions process prophylaxis after performance of operations on the abdominal cavity organs constitutes perspective trend for further investigations. PMID- 16719068 TI - [Peculiarities of diagnosis and surgical tactics in cystic pancreatic tumours]. AB - Results of selective analysis of examination and operative treatment data in patients with clinical diagnosis of cystic pancreatic tumour for ten-year period were presented. The causes of diagnostic and tactic mistakes were analyzed. The volume of necessary methods of instrumental and laboratory diagnosis, indications for operative treatment, volume of which depends on the tumour morphological characteristic, were determined. In serous cystadenoma the tumour excision or economic pancreatic resection is indicated; in mucinous tumour the extended pancreatic resection is indicated. PMID- 16719069 TI - [Anesthesiologic support of surgical intervention for diffuse purulent peritonitis]. AB - Original method of anesthesiological support of operative intervention in patients with diffuse purulent peritonitis is adduced. The method efficacy is confirmed by the results of studying of the central hemodynamics and metabolic component indices as well as the organism biological resistance. PMID- 16719070 TI - [Local signs of the reperfusion syndrome in obliterating atherosclerosis of the lower extremity arteries and their treatment]. AB - There were examined 32 patients with obliterating atherosclerosis of the lower extremities arteries, in whom the reperfusion syndrome have occurred after performance of the restoration operation. In 8 patients aorto-femoral shunting was performed and in 24--femoro-popliteal one. There was established that local signs of the reperfusion syndrome, as a rule, have occurred in patients with critical ischemia of the lower extremities preoperatively. Characteristic signs of the reperfusion syndrome were noted, the methods of its prophylaxis and treatment were adduced. PMID- 16719071 TI - [Method of surgical treatment of "elephantiasis" of the limb: old philosophy, new approach]. AB - The authors had summarized more than 25-year experience of surgical treatment of patients with the lower and upper extremities lymphedema. The aspiration resectional-plastic method of treatment of severe forms of the limb lymphedema, which intends in the first stage the resection of fibrously changed tissues, preserving the portion of slightly changed cutaneous-subcutaneous flaps and regions in projection of localization of main lymphatic collectors, under which aspiration lipectomy is performed. Postresectional wound surface is covered with special synthetic tissue. In the next stage, after appearance of granulation tissue, autodermoplasty is performed. Two-staged cutaneous plasty and repeated aspirational lipectomy provides for optimal survival of full thickness cutaneous transplants, reduction of the limb volume, improvement of the lymphatic system draining function. PMID- 16719072 TI - [Application of minimally invasive lumbar sympathectomy in the treatment of patients with obliterating diseases of the lower extremity vessels]. AB - The method of performance and immediate results of miniinvasive lumbar sympathectomy using retroperitoneal miniaccess were adduced in the work. In 2005 yr on the base of division of vascular surgery were operated 19 patients. Application of endoscopic instruments and optic system of illumination gave the possibility to remove lumbar sympathetic nodes in conditions of narrow (3-5 cm) surgical access and operative field. Introduction of miniinvasive method have promoted to reduce essentially the postoperative complications rate, the pain syndrome intensity, the reduction of duration of the patients postoperative rehabilitation, the cosmetic effect raising. PMID- 16719073 TI - [The external respiration function in patients with purulent-septic complications]. AB - In 7 patients with postoperative purulent-septic complications the external respiration function (ERF) was studied. Preoperatively in them were observed significant changes of the ERF speed characteristics. The average volume speed in 25-75% interval (AVS 25-75) of the forced vital pulmonary capacity (FVPC) was lower than normal level by 43%, the extreme volume speed (EVS) of FVPC--by 49%, the moment volume speed in inspiration 50% of FVPC (EVS 50)--by 42%, EVS in expiration 75% of FVPC (MOS 75)--by 48%. This have witnessed the pronounced obstructive pulmonary changes, which influenced negatively the postoperative period course. Insignificantly expressed restrictive changes did not cause the purulent-septic complications occurrence. PMID- 16719074 TI - [Laparoscopic interventions for adrenal gland tumor]. AB - Clinical course, hormonal characteristics and efficacy of rehabilitation were studied by the authors in 12 patients with suprarenal gland tumor, operated, using minimally invasive endoscopic intervention--laparoscopic adrenalectomy (LA). Efficacy, indications, contraindications, role of LA in the treatment of surgical adrenal diseases were analyzed. LA is characterized by invasiveness and traumaticity, adequate radicalism, quick rehabilitation, and good remote results. LA is indicated for the treatment of majority of hyperplastic and tumoral diseases of suprarenal glands: it is possible to operate 60% of patients with surgical diseases of suprarenal glands using endoscopic technology. PMID- 16719075 TI - [Combined cosmetic operation of the face]. AB - The method of performance of combined cosmetic operation in 58 women patients in 1996-2005 yrs. was analyzed. As a primary procedure for rejuvenation the combined operation of the face and neck skin lifting permits to achieve impressive results, prevents the "lateral bend" and hollow eyes formation. In patients with unfavourable consequences of the earlier performed standard face lifting the conduction of combined operation permits to eliminate them effectively. PMID- 16719076 TI - [Benign node in thyroid gland: "let's operate", "let's observe" or "let's try to treat"?]. PMID- 16719077 TI - [Severe course of gastroesophageal reflux disease caused by intra-abdominal tumoral compression]. PMID- 16719078 TI - The debate is over. PMID- 16719079 TI - Dead zones might masculinize fish. PMID- 16719080 TI - One-step process removes chromium from wastewater. PMID- 16719081 TI - Chicken poop and arsenic. PMID- 16719083 TI - The Weinberg proposal. PMID- 16719082 TI - Sorting out sources of perfluorinated chemicals. PMID- 16719084 TI - Toxin-free treated wood. PMID- 16719085 TI - Harnessing natural attenuation of pharmaceuticals and hormones in rivers. PMID- 16719086 TI - Modeling switchgrass derived cellulosic ethanol distribution in the United States. AB - Discussions of alternative fuel and propulsion technologies for transportation often overlook the infrastructure required to make these options practical and cost-effective. We estimate ethanol production facility locations and use a linear optimization model to consider the economic costs of distributing various ethanol fuel blends to all metropolitan areas in the United States. Fuel options include corn-based E5 (5% ethanol, 95% gasoline) to E16 from corn and switchgrass, as short-term substitutes for petroleum-based fuel. Our estimates of 1-2 cents per L of ethanol blend for downstream rail or truck transportation remain a relatively small fraction of total fuel cost. However, for even the relatively small blends of ethanol modeled, the transportation infrastructure demands would be comparably larger than the current demands of petroleum. Thus if ethanol is to be competitive in the long run, then in addition to process efficiency improvements, more efficient transportation infrastructure will need to be developed, such as pipelines. In addition to these results, national and regional policy challenges on how to pay for and optimize a new fuel and distribution infrastructure in the United States are discussed. PMID- 16719087 TI - N saturation symptoms in chaparral catchments are not reversed by prescribed fire. AB - Fire is a critical ecosystem process in many landscapes and is particularly dominant in the chaparral shrublands of southern California which are also exposed to high levels of atmospheric N deposition. Few studies have addressed the combined effects of these two disturbance factors. In this study we evaluate the hydrologic and biogeochemical response of a control and a prescribed burn catchment over a 15-year period. Streamwater nitrate concentrations and export in the burned catchment were higher than those in the unburned catchment for 7-10 years after the burn and concentrations remained high in both catchments during the entire study. Therefore, fire is not an effective mitigation tool for N deposition in these semi-arid systems. Additionally, the extended N export in this system indicates that chaparral ecosystems do not recovertheir N retention capabilities as rapidly as humid systems do when subjected to disturbance. PMID- 16719088 TI - Measurement of helium isotopes in soil gas as an indicator of tritium groundwater contamination. AB - The focus of this study was to define the shape and extent of tritium groundwater contamination emanating from a legacy burial ground and to identify vadose zone sources of tritium using helium isotopes (3He and 4He) in soil gas. Helium isotopes were measured in soil-gas samples collected from 70 sampling points around the perimeter and downgradient of a burial ground that contains buried radioactive solid waste. The soil-gas samples were analyzed for helium isotopes using rare gas mass spectrometry. 3He/4He ratios, reported as normalized to the air ratio (RA), were used to locate the tritium groundwater plume emanating from the burial ground. The 3He (excess) suggested that the general location of the tritium source is within the burial ground. This study clearly demonstrated the efficacy of the 3He method for application to similar sites elsewhere within the DOE weapons complex. PMID- 16719089 TI - Isomer distribution of perfluorocarboxylates in human blood: potential correlation to source. AB - Detection of perfluorocarboxylate anions (PFCAs), such as perfluorooctanoate (C7F15COO-, PFOA), at ng/g levels in human tissues has engendered public scrutiny of industrial fluorochemicals. Routes of PFCA exposure for the general human population are likely diverse given direct (industrially produced) and indirect (production from precursor organofluorines) sources. Major industrial production of organofluorines, including PFCAs, stems from either electrochemical fluorination (ECF) or telomerization. ECF products are a mixture of structural isomers (linear and branched perfluoroalkyls) and telomerization products are assumed to have one perfluorocarbon arrangement, typically linear. The objective of this research was to investigate structural isomer patterns of PFCAs in human blood. Volatile derivatives of PFCAs in human blood were analyzed by GC-(NCI)-MS for quantitation and isomers. PFOA was the dominant PFCA (mean 4.4 ng/g). Blood serum isomer profiles consisted of predominantly (mean approximately 98%) the linear isomer for each PFCA (C8-C11). There were similarities in branched isomer patterns of an ECF PFOA standard with both PFOA and PFNA in blood. Direct exposure to ECF PFOA, which has a legacy of production for uses in fluoropolymer industries, is postulated to be a source of the observed branched isomer pattern. Predominance of linear PFCA isomers and the [even PFCA] > [odd PFCA] concentration trend in blood is suggestive of additional input from a strictly linear perfluoroalkyl source. PMID- 16719090 TI - Sedimented organic nitrogen isotopes in freshwater wetlands record long-term changes in watershed nitrogen source and land use. AB - Although historic land use is widely recognized as an important determinant of watershed N cycling, efforts to examine land use legacy effects are limited by incomplete historical data. This research evaluates N isotopes of sedimented organic matter (delta15N(org)), in a palynological context, as a long-term proxy of changes in N source to wetland biota. N and S isotope measurements of organic sediments, fossil plant fragments, and living plants are used to explore isotope stratigraphies of wetland sediment cores. Processes potentially contributing to isotope stratigraphies are investigated including the following: a change in N source, diagenesis, and denitrification. We document the delta15N(org) stratigraphy of a core from the Smithsonian Environmental Research Center, MD, U.S.A. spans approximately 350 years, during which time delta15N(org) increases from +2 per thousand to +7 per thousand. Reconstructed population density and wastewater inputs to the watershed suggest that the increase in delta15N reflects changing land use from forested conditions to increasing nutrient inputs from human waste. Our results illustrate the importance of hydrologic connectivity in delivering waste-derived N in a watershed characterized by relatively low human population density. These results also demonstrate how this approach can expand the temporal horizon over which we can assess human impacts to watershed N dynamics. PMID- 16719091 TI - Monitoring algal toxins in lake water by liquid chromatography tandem mass spectrometry. AB - Microcystins (MCs) and cylindrospermopsin (CYL) are potent natural toxins produced by cyanobacteria (blue-green algae) that grow worldwide in eutrophic freshwaters and cause animal and human water-based toxicoses. The main purpose of this work has been assessing the contamination levels of some MCs and CYL in eutrophic Italian lake (Albano) water. To do this, we have developed an original analytical method involving MC extraction with a sorbent (Carbograph 4) cartridge. CYL is a highly polar compound that is scarcely retained by any sorbent material. To analyze this toxin, we directly injected 0.5 mL of filtered lake water into the liquid chromatography (LC) column. Analytes were quantified by LC coupled to tandem mass spectrometry in the multireaction monitoring mode. The recovery of five selected MCs added to an analyte free lake water sample at three different concentrations (50, 150, and 500 ng/L) ranged between 93 and 103% with RSD values no larger than 8%. Limits of quantification (LOQ) of the five MCs were within the 2-9 ng/L range, whereas the LOQ of CYL was 300 ng/L. The occurrence and abundance of cyanotoxins in Lake Albano was monitored over four months (Sept-Dec 2004) by analyzing water samples collected monthly at the center of the lake and at different depths (from 0 to -30 m). During survey and with the MS/MS system operating in the parent ion scan mode, we individuated two demethylated forms of MC-RR and one demethylated variety of MC-LR. Demethylated MC-RRs are known to be even more toxic than MC-RR toward zooplanktic grazers. CYL was the most-abundant toxin during the first three monitoring months. To the best of our knowledge, this is the first work reporting concentration levels of CYL in lake water. PMID- 16719092 TI - Health risks in infants associated with exposure to perfluorinated compounds in human breast milk from Zhoushan, China. AB - Recent studies have reported the ubiquitous distribution of perfluorinated compounds (PFCs), especially perfluorooctanesulfonate (PFOS) and perfluorooctanoic acid (PFOA), in wildlife and human whole blood or serum. In 2003 a solid phase extraction method was developed, which allowed the measurement of PFCs in human breast milk. In the present study, PFCs in samples of human breast milk from 19 individuals from Zhoushan, China, were analyzed by modifying a previously established method, based on weak-anion exchange extraction. PFOS and PFOA were the two dominant chemicals detected in all the milk samples. Concentrations of PFOS and PFOA ranged from 45 to 360 ng/L and 47 to 210 ng/L, respectively. The maximum concentrations of other PFCs were 100 ng/L for perfluorohexanesulfonate (PFHxS), 62 ng/L for perfluorononanoate (PFNA), 15 ng/L for perfluorodecanoate (PFDA) and 56 ng/L for perfluoroundecanoate (PFUnDA). Statistically significant correlations between various PFCs suggested a common exposure source to humans. No statistically significant correlation was found between concentrations of either PFOS or PFOA and maternal age, weight, or infant weight. Rate of consumption of fish was found to be positively correlated with PFNA, PFDA, and PFUnDA concentrations. Daily intake of PFOS for the child via breast milk with greater PFOS concentrations exceeded the predicted conservative reference dose in 1 of 19 samples, indicating that there may be a small potential risk of PFOS for the infants in Zhoushan via the consumption of breast milk. PMID- 16719093 TI - Bioaccumulation, biotransformation, and metabolite formation of fipronil and chiral legacy pesticides in rainbow trout. AB - To assess the fate of current-use pesticides, it is important to understand their bioaccumulation and biotransformation by aquatic biota. We examined the dietary accumulation and enantioselective biotransformation of the chiral current-use pesticide fipronil, along with a mixture of selected chiral [alpha hexachlorocyclohexane (alpha-HCH), heptachlor epoxide (HEPX), polychlorinated biphenyls (PCBs) 84, 132, 174, o,p'-DDT, and o,p'-DDD] and nonchiral (p,p'-DDT, p,p'-DDD) organochlorine compounds in juvenile rainbow trout (Oncorhynchus mykiss). Fish rapidly accumulated all compounds, as measured in the carcass (whole body minus liver and GI tract) during the 32 d uptake phase, which was followed by varying elimination rates of the chemicals (half-lives (t1/2s) ranging from 0.6 d for fipronil to 77.0 d for PCB 174) during the 96 d depuration period. No biotransformation was observed for alpha-HCH, HEPX, PCB 174, o,p'-DDT, or o,p'-DDD based on consistent enantiomeric fractions (EFs) in the fish and their t1/2s falling on a log K(ow)--log t1/2 relationship established for recalcitrant contaminants in fish. p,p'-DDT and PCBs 84 and 132 were biotransformed based on the former's t1/2 position below the log K(ow)--log t1/2 relationship, and the PCBs change in EF. Fipronil was rapidly biotransformed, based on a change in EF, a t1/2 that fell below the log K(ow)--log t1//2 relationship, which accounted for 88% of its elimination, and the rapid formation of fipronil sulfone, a known metabolite. Fipronil sulfone was found to persist longer (t1/2 approximately 2 d) than its parent compound fipronil (t1/2 approximately 0.6 d) and needs to be considered in fate studies of fipronil. This research demonstrates the utilities of the log K(ow)--log t1/2 relationship as a mechanistic tool for quantifying biotransformation and of chiral analysis to measure biotransformation in fish. PMID- 16719094 TI - Remarkable findings concerning PBDEs in the terrestrial top-predator red fox (Vulpes vulpes). AB - In the present study, we have analyzed muscle, liver, and adipose tissue of 33 red foxes from Belgium for their content of polybrominated diphenyl ethers (PBDEs). Median sums of seven tri- to hepta-BDEs (BDE 28, BDE 47, BDE 99, BDE 100, BDE 153, BDE 154, and BDE 183) were 2.2, 2.4, and 3.4 ng/g lipid weight in adipose tissue, liver, and muscle, respectively. These levels were lower than those found in various species of voles and mice, the main prey species of the red fox. This is probably related to the high capacity of the foxes to metabolize and eliminate lower brominated congeners. BDE 209 generally dominated the PBDE congener profiles in the red fox samples. In samples containing BDE 209, this congener contributed, on the average, approximately 70% to the total PBDE content. BDE 209 was measured in concentrations as high as 760 ng/g lipid weight in the liver, but the detection frequency was not more than 40%. In animals with the highest BDE 209 levels, this congener was detected in muscle, liver, as well as in adipose tissue. Other abundant congeners were BDE 153 and BDE 47, which prevail in other terrestrial species. The particular PBDE congener profile observed in the red fox resembles that seen in grizzly bears from Canada, but differs from those previously reported for terrestrial avian species. Our data confirms unambiguously that BDE 209 does bioaccumulate in terrestrial top predators, such as the red fox. PMID- 16719095 TI - Experimentally altered groundwater inflow remobilizes acidity from sediments of an iron rich and acidic lake. AB - To study the impact of changes in groundwater flow and chemistry on acidity export from sediments in acid mine drainage (AMD) polluted lakes, a column experiment was carried out. Schwertmannite rich sediment was subjected to three different flow rates (0, 5, and 20 L m(-2) a(-1)), two percolate chemistries (1/1 mmol L(-1) vs 10/15 mmol L(-1) sulfate/ferrous iron, pH 5), and DOC input (approximately 2.5 mmol C L(-1)). Percolation induced acidity export in all percolated treatments (8.8-40.4 mol m(-2) a(-1)) by accelerated proton generation from schwertmannite transformation (18.0-35.9 mol m(-2) a(-1)) and ferrous iron release (3.8-11.6 mol m(-2) a(-1)) from the sediment matrix. Mobilization increased with flow rate and decreased with sulfate and iron concentrations. Unspecifically bound ferrous iron contents increased within the sediment (up to 40.5 mol m(-2) a(-1)) when iron concentrations in the percolate were high. Reduced sulfur species formed following raises in pH, but acidity consumption through this process (0.3-6.6 mol m(-2) a(-1)) and the formation of carbonates (0.11-0.45 mol m(-2) a(-1)) remained small. The study thus suggests that increases in groundwater inflow remobilize acidity from AMD polluted sediments. PMID- 16719096 TI - Anaerobic biotransformation of roxarsone and related N-substituted phenylarsonic acids. AB - Large quantities of arsenic are introduced into the environment through land application of poultry litter containing the organoarsenical feed additive roxarsone (3-nitro-4-hydroxyphenylarsonic acid). The objective of this study was to evaluate the bioconversion of roxarsone and related N-substituted phenylarsonic acid derivatives under anaerobic conditions. The results demonstrate that roxarsone is rapidly transformed in the absence of oxygen to the corresponding aromatic amine, 4-hydroxy-3-aminophenylarsonic acid (HAPA). The formation of HAPA is attributable to the facile reduction of the nitro group. Electron-donating substrates, such as hydrogen gas, glucose, and lactate, stimulated the rate of nitro group reduction, indicating a microbial role. During long-term incubations, HAPA and the closely related 4-aminophenylarsonic acid (4 APA) were slowly biologically eliminated by up to 99% under methanogenic and sulfate-reducing conditions, whereas little or no removal occurred in heat-killed inoculum controls. Arsenite and, to a lesser extent, arsenate were observed as products of the degradation. Freely soluble forms of the inorganic arsenical species accounted for 19-28% of the amino-substituted phenylarsonic acids removed. This constitutes the first report of a biologically catalyzed rupture of the phenylarsonic group under anaerobic conditions. PMID- 16719097 TI - Absorption and adsorption of hydrophobic organic contaminants to diesel and hexane soot. AB - Soot particles vary in pore structure, surface properties, and content of authigenic (native) extractable organic chemicals. To better understand the effects of these properties on sorption, aqueous sorption isotherms for 14C labeled phenanthrene and 1,2,4-trichlorobenzene were obtained for four soots of varying properties: two diesel reference soots, a hexane soot, and an ozonated hexane soot. Substantial isotherm nonlinearity was observed. In comparison to diesel soot SRM 2975, diesel soot SRM 1650b had a much higher content of extractable authigenic organic chemicals, showed less sorption of 14C-labeled sorbate at low relative concentrations (Ce/Sw), and showed higher sorption at high Ce/Sw. In comparison to normal hexane soot, the ozonated hexane soot had a higher surface O/C ratio and showed substantially less sorption at all concentrations studied. The sorption differences were attributed to the noted differences in properties, and results were interpreted through a dual-mode sorption model that included the possibility of both surface adsorption (modeled using a Polanyi-based approach) and simple phase partitioning (linear absorption). Generally, such modeling indicated that overall uptake at low concentrations in all four soots was dominated by surface adsorption but that sorption at higher sorbate concentrations in SRM 1650b was heavily influenced by linear absorption within the natively bound organic phase. PMID- 16719098 TI - Regional patterns in the isotopic composition of natural and anthropogenic nitrate in groundwater, High Plains, U.S.A. AB - Mobilization of natural nitrate (NO3-) deposits in the subsoil by irrigation water in arid and semiarid regions has the potential to produce large groundwater NO3-concentrations. The use of isotopes to distinguish between natural and anthropogenic NO3- sources in these settings could be complicated by the wide range in delta15N values of natural NO3-. An approximately 10 000 year record of paleorecharge from the regionally extensive High Plains aquifer indicates that delta15N values for NO3- derived from natural sources ranged from 1.3 to 12.3 per thousand and increased systematically from the northern to the southern High Plains. This collective range in delta15N values spans the range that might be interpreted as evidence for fertilizer and animal-waste sources of NO3-; however, the delta15N values for NO3- in modern recharge (< 50 years) under irrigated fields were, for the most part, distinctly different from those of paleorecharge when viewed in the overall regional context. An inverse relation was observed between the delta15N[NO3-] values and the NO3-/Cl- ratios in paleorecharge that is qualitatively consistent with fractionating losses of N increasing from north to south in the High Plains. N and O isotope data for NO3- are consistent with both NH3 volatilization and denitrification, having contributed to fractionating losses of N prior to recharge. The relative importance of different isotope fractionating processes may be influenced by regional climate patterns as well as by local variation in soils, vegetation, topography, and moisture conditions. PMID- 16719099 TI - Interactions of DNA with clay minerals and soil colloidal particles and protection against degradation by DNase. AB - Adsorption, desorption, and degradation by nucleases of DNA on four different colloidal fractions from a Brown soil and clay minerals were studied. The adsorption of DNase I and the structures of native DNA, adsorbed and desorbed, were also investigated by Fourier Transform Infrared (FTIR), circular dichroism (CD), and fluorescence spectroscopy, to determine the protection mechanism of DNA molecules by soil colloids and minerals against enzymatic degradation. Kaolinite exhibited the highest adsorption affinity for DNA among the examined soil colloids and clay minerals. In comparison with organomineral complexes (organic clays), DNA was tightly adsorbed by H2O2-treated clays (inorganic clays). FTIR spectra showed that the binding of DNA on kaolinite and inorganic clays changed its conformation from the B-form to the Z-form, whereas montmorillonite and organic clays retained the original B-form of DNA. A structural change from the B to the C-form in DNA molecules desorbed from kaolinite was observed by CD spectroscopy and confirmed by fluorescence spectroscopy. The presence of soil colloids and minerals provided protection to DNA against degradation by DNase I. The higher level of protection was found with montmorillonite and organic clays compared to kaolinite and inorganic clays. The protection of DNA against nuclease degradation by soil colloids and minerals is apparently not controlled by the adsorption affinity of DNA molecules for the colloids and the conformational change of bound DNA. The higher stability of DNA seemed to be attributed mainly to the presence of organic matter in the system and the adsorption of nucleases on soil colloids and minerals. The information obtained in this study is of fundamental significance for the understanding of the behavior of extracellular DNA in soil environment. PMID- 16719100 TI - Chloride retention and release in a boreal forest soil: effects of soil water residence time and nitrogen and chloride loads. AB - The common assumption that chloride (Cl-) is conservative in soils and can be used as a groundwater tracer is currently being questioned, and an increasing number of studies indicate that Cl- can be retained in soils. We performed lysimeter experiments with soil from a coniferous forest in southeast Sweden to determine whether pore water residence time and nitrogen and Cl- loads affected Cl- retention. Over the first 42 days there was a net retention of Cl- with retention rates averaging 3.1 mg CI- m(-2) d(-1) (68% of the added Cl- retained over 42 days). Thereafter, a net release of Cl- at similar rates was observed for the remaining experimental period (85 d). Longer soil water residence time and higher Cl- load gave higher initial retention and subsequent release rates than shorter residence time and lower Cl- load did. Nitrogen load did not affect Cl transformation rates. This study indicates that simultaneous retention and release of Cl- can occur in soils, and that rates may be considerable relative to the load. The retention of Cl- observed was probably due to chlorination of soil organic matter or ion exchange. The cause of the shift between net retention and net release is unclear, but we hypothesize that the presence of O2 or the presence of microbially available organic matter regulates Cl- retention and release rates. PMID- 16719101 TI - Localized attraction correlates with bacterial adhesion to glass and metal oxide substrata. AB - Bacterial adhesion to surfaces does not always proceed according to theoretical expectations. Discrepancies are often attributed to surface heterogeneities that provide localized, favorable sites for bacterial attachment. The presence of these favorable deposition sites for bacteria, however, has never been directly measured. Atomic force microscopy (AFM) was used to quantify the distribution of attractive sites on clean substrata. Surfaces of silica and three different metal oxides mapped by adhesion force with regular or colloidal AFM tips showed a heterogeneous distribution of adhesion forces. Adhesion forces were normally distributed based on a colloid probe, but regular tips revealed a proportionately larger number of relatively more adhesive sites. No correlation was found between the average adhesion force (tip or colloid) and macroscopic adhesion tests using five strains of bacteria. However, when AFM tip results were compared to bacterial adhesion data on the basis of only the stickiest sites (the 5% of sites with the largest adhesion force), there was a good correlation of AFM data with adhesion data. These results demonstrate for the first time how overall bacterial adhesion to a surface effectively correlates with a relatively small fraction of highly adhesive sites rather than averaged adhesion force as detected using AFM. PMID- 16719102 TI - Phototransformation of the herbicide sulcotrione on maize cuticular wax. AB - Vegetation plays a key role in environmental cycling and the fate of many organic pollutants. This is especially the case for pesticides because plant leaves are their first reaction environment after application. It is commonly accepted that photochemical reactions of pollutants on plants predominantly take place in the cuticular wax coating of the leaves. Thus, we used films made of either cuticular wax extracted from maize or carnauba gray wax as a model support. Under simulated sunlight irradiation, sulcotrione (a new class of triketone herbicides) sorbed on cuticular wax films was photolyzed and mainly underwent an intramolecular cyclization. The photoproduct is a chromone derivative which was isolated and fully characterized. It is reported for the first time as a sulcotrione degradation product. The photoreactivity of formulated sulcotrione at the surface of cuticular waxes was investigated too. It photodegraded more rapidly than nonformulated sulcotrione. This study also shows that the rate of sulcotrione photolysis was much faster than the rate of penetration into the wax; photolysis should be, thus, a relevant process in real conditions. PMID- 16719103 TI - Trans boundary transport of pollutants by atmospheric mineral dust. AB - The transport of anthropogenic pollution by desert dust in the Eastern Mediterranean region was studied by analyzing major and trace element composition, organic species, and Pb isotope ratios in suspended dust samples collected in Jerusalem, Israel. Dust storms in this region are associated with four distinct synoptic conditions (Red Sea Trough (RS), Eastern High (EH), Sharav Cyclone (SC), and Cold Depression (Cyprus low, CD)) that carry dust mostly from North African (SC, CD, EH) and Arabian and Syrian (RS, EH) deserts. Substantial contamination of dust particles by Pb, Cu, Zn, and Ni is observed, while other elements (Na, Ca, Mg, Mn, Sr, Rb, REE, U, and Th) display natural concentrations. Sequential extraction of the abovementioned elements from the dust samples shows that the carbonate and sorbed fractions contain most of the pollution, yet the Al silicate fraction is also contaminated, implying that soils and sediments in the source terrains of the dust are already polluted. We identified the pollutant sources by using Pb isotopes. It appears that before the beginning of the dust storm, the pollutants in the collected samples are dominated by local sources but with the arrival of dust from North Africa, the proportion of foreign pollutants increases. Organic pollutants exhibit behavior similar and complementary to that of the inorganic tracers, attesting to the importance of anthropogenic-pollutant addition en route of the dust from its remote sources. Pollution of suspended dust is observed under all synoptic conditions, yet it appears that easterly winds carry higher proportions of local pollution and westerly winds carry pollution emitted in the Cairo basin. Therefore, pollution transport by mineral dust should be accounted for in environmental models and in assessing the health related effects of mineral dust. PMID- 16719105 TI - SOA formation from partitioning and heterogeneous reactions: model study in the presence of inorganic species. AB - A predictive model for secondary organic aerosol (SOA) formation by both partitioning and heterogeneous reactions was developed for SOA created from ozonolysis of alpha-pinene in the presence of preexisting inorganic seed aerosols. SOA was created in a 2 m3 polytetrafluoroethylene film indoor chamber under darkness. Extensive sets of SOA experiments were conducted varying humidity, inorganic seed compositions comprising of ammonium sulfate and sulfuric acid, and amounts of inorganic seed mass. SOA mass was decoupled into partitioning (OM(P)) and heterogeneous aerosol production (OM(H)). The reaction rate constant for OM(H) production was subdivided into three categories (fast, medium, and slow) to consider different reactivity of organic products for the particle phase heterogeneous reactions. The influence of particle acidity on reaction rates was treated in a previous semiempirical model. Model OM(H) was developed with medium and strong acidic seed aerosols, and then extrapolated to OM(H) in weak acidic conditions, which are more relevant to atmospheric aerosols. To demonstrate the effects of preexisting glyoxal derivatives (e.g., glyoxal hydrate and dimer) on OM(H), SOA was created with a seed mixture comprising of aqueous glyoxal and inorganic species. Our results show that heterogeneous SOA formation was also influenced by preexisting reactive glyoxal derivatives. PMID- 16719104 TI - Abiotic reduction of nitroaromatic compounds by aqueous iron(ll)-catechol complexes. AB - Complexation of iron(ll) by catechol and thiol ligands leads to the formation of aqueous species that are capable of reducing substituted nitroaromatic compounds (NACs) to the corresponding anilines. No reactions of NACs are observed in FelI only or ligand-only solutions. In solutions containing FeII and tiron, a model catechol, rates of NAC reduction are heavily dependent on pH, ligand concentration, and ionic strength. Observed pseudo-first-order rate constants (k(obs)) for 4-chloronitrobenzene reduction vary by more than 6 orders of magnitude, and the variability is well described by the expression k(obs) = k(FeL2)(6-) [FeL2(6-)], where [FeL2(6-)] is the concentration of the 1:2 FeII tiron complex and kFeL2(6-) is the bimolecular rate constant for 4 chloronitrobenzene reaction with this species. The high reactivity of this FeII species is attributed to the low standard one-electron reduction potential of the corresponding FeIII/FeII redox couple (EH0 = -0.509 V vs NHE). The relative reactivity of different NACs can be described by a linear free-energy relationship (LFER) with the one-electron reduction potentials of the NACs, EH1'(ArNO2). The experimentally derived slope of the LFER indicates that electron transfer is rate determining. These findings suggest that FeII-organic complexes may play an important, previously unrecognized, role in the reductive transformation of persistent organic contaminants. PMID- 16719106 TI - Monofluorinated analogues of polybrominated diphenyl ethers as analytical standards: synthesis, NMR, and GC-MS characterization and molecular orbital studies. AB - Polybrominated diphenyl ethers (PBDEs), a group of 209 individual congeners distinguishable by the number and position of bromines, are produced for use as flame retardants in consumer goods. PBDEs have become ubiquitous environmental contaminants, present in increasing levels in the environment and humans. In the present study, 10 individual monofluorinated analogues of PBDEs (F-PBDEs) and one difluorinated PBDE (FF-PBDE) were synthesized and characterized, and their gas chromatographic (GC) and mass spectrometric (MS) characteristics determined. The synthesis method utilized a nucleophilic reaction of bromophenols with diphenyliodonium salts and the perbromination of fluorosubstituted diphenyl ethers. Reaction yields were between 10% and 59% with > or = 98% purity. Apart from the aromatic ring carrying the fluorine atom, only minor chemical nuclear magnetic resonance (NMR) shift changes were observed in comparison to the corresponding parent PBDEs, with the exception that the J(F,H) coupling was stronger. Our preliminary data show that F-PBDEs and PBDEs have comparable retention times in gas chromatography with F-PBDEs demonstrating in general shorter or identical retention times, depending on the pattern of fluorine substitution. We also calculated the torsion angles and the dipole moments for both and report that there is a good correlation between GC retention times and the torsion angles but not with dipole moments. In MS, the difference of the ion peaks of the F-PBDE/ PBDE pairs is m/z 19 (F), which allows a simultaneous MS detection without separation. On the basis of GC separation, simultaneous MS detection, and the stability of fluorine due to its generally resistance to nucleophilic displacement, we propose that F-PBDEs may function as valuable potential standards, markers, and tracers in environmental analysis. PMID- 16719107 TI - Mitigation of ammonia and hydrogen sulfide emissions by stable aqueous foam microbial media. AB - Stable aqueous foam-microbial media consisting of protein-based foams and odor degrading bacteria were developed to control the emissions of odorous compounds. The optimum foam formulation was determined based on foam characteristics including 50% drainage time, foam lifetime, and foam expansion ratio. When only the aqueous foam was applied onto the surface of a test odor source (i.e., swine manure), ammonia emission was completely suppressed for about 177, 225, 265, 297, and 471 min when the height of foam barrier was 2.5, 5, 10, 15, and 30 cm, respectively. According to the increasing foam height, ammonia emission rates after breakthrough points decreased to 0.16, 0.13, 0.09, 0.07, and 0.02 mg/m3/min, and thus volatilized ammonia concentrations decreased significantly after 600 min. Hydrogen sulfide was similarly suppressed. Ammonia emission was better controlled by incorporating odor-degrading bacteria into the aqueous foam. The odor suppression capacity of the 5-cm foam barrier with microbes was more than eight times greater than that of the barrier only and was similar to that of 30-cm foam barrier without microbes after 1440 min. A significant amount of dinitrogen gas was evolved by the foam-microbial media, indicating a successful biological transformation of ammonia. PMID- 16719108 TI - Formation of bromochlorodibenzo-p-dioxins and dibenzofurans from the high temperature oxidation of a mixture of 2-chlorophenol and 2-bromophenol. AB - The homogeneous, gas-phase oxidative thermal degradation of a 50:50 mixture of 2 bromophenol and 2-chlorophenol was studied in a 1 cm i.d., fused silica flow reactor at a concentration of 88 ppm, with a reaction time of 2.0 s, over a temperature range of 300 to 1000 degrees C. Observed products in order of decreasing yield included the following: dibenzo-p-dioxin (DD), 4-bromo-6 chlorodibenzofuran (4-B,6-CDF), phenol, 4,6-dibromodibenzofuran (4,6-DBDF), 2,6 dibromophenol, 4,6-dichlorodibenzofuran (4,6-DCDF), 2-bromo-4-chlorophenol, 2,4 dibromophenol, 2-chloro-4-bromophenol, 4-monobromodibenzofuran (4-MBDF), 4 monochlorodibenzofuran (4-MCDF), dibenzofuran (DF), 1-monobromodibenzo-p-dioxin (1-MBDD), 1-monochlorodibenzo-p-dioxin (1-MCDD), 2,4,6-tribromophenol, naphthalene, chloronaphthalene, bromonaphthalene, chlorobenzene, bromobenzene, and benzene. The results are compared and contrasted with previous results reported for the oxidations of pure 2-chlorophenol and 2-bromophenol as well as results for the pyrolysis of the mixture of 2-chlorophenol and 2-bromophenol. 4,6 DBDF and 4,6-DCDF were observed in higher yields than under pyrolytic conditions but considerably less than the yields observed for the individual oxidation of 2 chlorophenol and 2-bromophenol. The effect on chlorine and bromine on the concentration of hydroxyl radical is shown to control the dioxin-to-furan ratio. PMID- 16719109 TI - Dechlorinating chloroacetanilide herbicides by dithionite-treated aquifer sediment and surface soil. AB - The prevalent use of chloroacetanilide herbicides has resulted in nonpoint contamination of some groundwater and surface water. We determined the efficacy of dithionite-treated sediment and soils to transform chloroacetanilides. When used alone, dithionite rapidly dechlorinates chloroacetanilides in water, with the following order of reactivity: propachlor > alachlor > acetochlor > metolachlor. Stoichiometric release of chloride occurs during reaction with dithionite, and thiosulfate herbicide derivatives are produced. Treating aquifer sediment with dithionite reduces native Fe(lII), creating a redox barrier of Fe(ll)-bearing minerals and surface-bound Fe(ll). Washing the reduced sediment (buffered with citrate-bicarbonate) with oxygen-free water removed Fe(ll) and excess dithionite and no alachlor transformation was observed. In contrast, a dithionite-treated surface soil, rich in clay and iron, effectively dechlorinated alachlor after washing. Exposing alachlor to aquifer sediment treated with dithionite in potassium carbonate buffer (pH 8.5-9.0) produced dechlorinated alachlor as the major degradation product. Our results provide proof-of-concept that dechlorination of chloroacetanilide herbicides by dithionite and dithionite treated aquifer sediment and soil is a remediation option in natural environments where iron-bearing minerals are abundant. PMID- 16719110 TI - Effect of bromide and iodide ions on the formation and speciation of disinfection byproducts during chlorination. AB - Two natural waters were fortified with various levels of bromide or iodide ions (0-30 microM) and chlorinated in the laboratory to study the impact of bromide and iodide ions on the formation and speciation of disinfection byproducts. Trihalomethanes (THMs), haloacetic acids (HAAs), total organic halogen (TOX), and its halogen-specific fractions total organic chlorine (TOCl), bromine (TOBr), and iodine (TOI), were measured in this work. The molar yields of THMs and HAAs increased as the initial bromide concentration increased. No significant change in TOX concentration was found for varying bromide concentrations. However, TOX concentrations decreased substantially with increasing initial iodide concentrations. At higher levels of bromide, there was a decreasing level of unknown TOX and unknown TOCl but an increasing level of unknown TOBr. The extent of iodine substitution was much lower than that of bromine substitution when comparing identical initial concentrations because a substantial amount of iodide was oxidized to iodate by chlorine. The tendency toward iodate formation resulted in the unusual situation where higher chlorine doses actually caused reduced levels of iodinated organic byproducts. Quantitative assessment of the results of this study showed a good agreement with kinetic data in the literature. PMID- 16719111 TI - Phenols and amine induced HO* generation during the initial phase of natural water ozonation. AB - The initial phase of ozone decomposition in natural water (t < 20 s) is poorly understood. It has recently been shown to result in very high transient HO* concentrations and, thereby, plays an essential role during processes such as bromate formation or contaminants oxidation. Phenols and amines are ubiquitous moieties of natural organic matter. Naturally occurring concentrations of primary, secondary, and tertiary amines, amino acids, and phenol were added to surface water, and ozone decomposition as well as HO* generation were measured starting 350 milliseconds after ozone addition. Six seconds into the process, 5 microM of dimethylamine and phenol had generated integral of HO* dt = 1 x 10(-10) M*s and 1.8 x 10(-10) M*s, respectively. With 10 microM dimethylamine and 1.5 mg O3/L, R(ct), (integral of HO*dt/ integral of O3dt) reached 10(-6), which is larger than in advanced oxidation processes (AOP) such as O3/H2O2. Experiments in the presence of HO*-scavengers indicated that a significant fraction of phenol induced ozone decomposition and HO* generation results from a direct electron transfer to ozone. For dimethylamine, the main mechanism of HO* generation is direct formation of O2*- which reacts selectively with O3 to form O3*-. Pretreatment of phenol-containing water with HOCl or HOBr did not decrease HO* generation, while the same treatment of dimethylamine-containing water considerably reduced HO* generation. PMID- 16719114 TI - Effects of natural water ions and humic acid on catalytic nitrate reduction kinetics using an alumina supported Pd-Cu catalyst. AB - Catalytic nitrate reduction was evaluated for the purpose of drinking water treatment. Common anions present in natural waters and humic acid were evaluated for their effects on NO3(-) hydrogenation over a bimetallic supported catalyst (Pd-Cu/gamma-Al2O3). Groundwater samples, with and without powder activated carbon (PAC) pretreatment, were also evaluated. In the absence of inhibitors the NO3- reduction rate was 2.4 x 10(-01) L/min g cat. However, the addition of constituents (SO4(2-), SO3(2-), HS-, CI-, HCO3-, OH-, and humic acid) on the order of representative concentrations for drinking water decreased the NO3- reduction rate. Sulfite, sulfide, and elevated chloride decreased the NO3- reduction rate by over 2 orders of magnitude. Preferential adsorption of Cl- inhibited NO3- reduction to a greater extent than NO2- reduction. Partial regeneration of catalysts exposed to SO3(2-) was achieved by using a dilute hypochlorite solution, however Cu dissolution occurred. Dissolved constituents in the groundwater sample decreased the NO3- reduction rate to 3.7 x 10(-03) L/min g cat and increased ammonia production. Removal of dissolved organic matter from the groundwater using PAC increased the NO3- reduction rate to 5.06 x 10(-02) L/min g cat and decreased ammonia production. Elemental analyses of catalysts exposed to the natural groundwater suggest that mineral precipitation may also contribute to catalyst fouling. PMID- 16719113 TI - Removal of toxic uranium from synthetic nuclear power reactor effluents using uranyl ion imprinted polymer particles. AB - Major quantities of uranium find use as nuclear fuel in nuclear power reactors. In view of the extreme toxicity of uranium and consequent stringent limits fixed by WHO and various national governments, it is essential to remove uranium from nuclear power reactor effluents before discharge into environment. Ion imprinted polymer (IIP) materials have traditionally been used for the recovery of uranium from dilute aqueous solutions prior to detection or from seawater. We now describe the use of IIP materials for selective removal of uranium from a typical synthetic nuclear power reactor effluent. The IIP materials were prepared for uranyl ion (imprint ion) by forming binary salicylaldoxime (SALO) or 4 vinylpyridine (VP) or ternary SALO-VP complexes in 2-methoxyethanol (porogen) and copolymerizing in the presence of styrene (monomer), divinylbenzene (cross linking monomer), and 2,2'-azobisisobutyronitrile (initiator). The resulting materials were then ground and sieved to obtain unleached polymer particles. Leached IIP particles were obtained by leaching the imprint ions with 6.0 M HCl. Control polymer particles were also prepared analogously without the imprint ion. The IIP particles obtained with ternary complex alone gave quantitative removal of uranyl ion in the pH range 3.5-5.0 with as low as 0.08 g. The retention capacity of uranyl IIP particles was found to be 98.50 mg/g of polymer. The present study successfully demonstrates the feasibility of removing uranyl ions selectively in the range 5 microg - 300 mg present in 500 mL of synthetic nuclear power reactor effluent containing a host of other inorganic species. PMID- 16719112 TI - Removal of Cr(VI) and Cr(lll) from aqueous solutions and industrial wastewaters by natural clino-pyrrhotite. AB - This paper introduced a simple method of treating Cr(Vl)-bearing toxic wastewaters using a natural mineral: clino-pyrrhotite. Laboratory bench-scale mixing experiments were carried out in both Cr(VI)-bearing artificial solutions and industrial wastewaters under controlled conditions. The effects of solution pH, Cr(VI) concentration, mineral grain size, mineral/solution ratio, and reaction time on the Cr(VI) removal were studied. Chromium was effectively removed from the solutions and wastewaters. After the treatment, the liquid was clean enough to be discharged directly into the natural environment. The Cr(VI) removal process involved sequentially the adsorption of Cr(VI), in the form of Cr2O7(2-) or CrO4(2-), onto the mineral surface, the reduction of the adsorbed Cr(VI) to Cr(lll), catalyzed at the vacant Fe sites of the mineral, and finally the precipitation of Cr(lll) as Cr2S3, Cr2O3, and Cr(OH)3 solid phases. Conditions such as a fine mineral grain size, an excessive quantity of clino pyrrhotite and a weak acidic media, favored the removal process. For clino pyrrhotite with a restricted grain size, the minimum required quantity of the mineral was proportional to the total quantity of Cr(VI) to be removed. Quantitatively, one cubic meter of industrial wastewater that contained approximately 1 mmol dm(-3) of Cr(VI) and had a pH value between 1 and 10 would be effectively treated after it was in contact with 220 kg of 145 +/- 28 microm clino-pyrrhotite for an hour. Furthermore, the quantity of the final solid waste byproduct was small, and the solid residue of clino-pyrrhotite could be reused after a simple rinse with water. Compared to the previous Cr(VI)-bearing wastewater treatment schemes, this method was simple, effective, economical, and environmentally sound. It has great potential for use in industrial-scale applications. PMID- 16719115 TI - Ionizing radiation-induced destruction of benzene and dienes in aqueous media. AB - Pulse radiolysis with spectrophotometric and conductometric detection was utilized to study the formation and reactions of radicals from benzene and dienes in aqueous solutions. The benzene OH adduct, *C6H6OH, reacts with O2 (k = 3 x 10(8) L mol(-1) s(-1)) in a reversible reaction. The peroxyl radical, HOC6H6O2*, undergoes O2*- elimination, bimolecular decay, and reaction with benzene to initiate a chain reaction, depending on the dose rate, benzene concentration, and pH. The occurrence of the chain reaction is demonstrated in low-dose-rate gamma radiolysis experiments where the consumption of O2 was monitored. 1,4 Cyclohexadiene, 1,4-hexadiene, and 1,4-pentadiene form OH-adducts and undergo H abstraction by O*- radicals. The OH-adducts react with O2 to form peroxyl radicals. These peroxyl radicals, however, do not undergo unimolecular O2*- elimination but rather decay by second-order processes, which lead to subsequent steps of O2*- elimination. PMID- 16719116 TI - Biofiltration and inhibitory interactions of gaseous benzene, toluene, xylene, and methyl tert-butyl ether. AB - This study evaluated the individual and combined removal capacities of benzene, toluene, and xylene (B, T, and X) in the presence and absence of methyl tert butyl ether (MTBE) in a polyurethane biofilter inoculated with a BTX-degrading microbial consortium, and further examined their interactive effects in various mixtures. In addition, Polymerase chain reaction-denaturing gradient gel electrophoresis and phylogenetic analysis of 16S rRNA gene sequences were used to compare the microbial community structures found in biofilters exposed to the various gases and gas mixtures. The maximum individual elimination capacities (MECs) of B, T, and X were 200, 238, and 400 g m(-3) h(-1), respectively. There was no significant elimination of MTBE alone. Addition of MTBE decreased the MECs of B,T, and X to 75, 100, and 300 g m(-3) h(-1), respectively, indicating that benzene was most strongly inhibited by MTBE. When the three gases were mixed (B + T + X), the removal capacities of individual B, T, and X were 50, 90, and 200 g m(-3) h(-1), respectively. These capacities decreased to 40, 50, and 100 g m(-3) h(-1) when MTBE was added to the mix. The MEC of the three-gas mixture (B + T + X) was 340 g m(-3) h(-1), and that of the four-gas mixture was 200 g m(-3) h(-1). Although MTBE alone was not degraded by the biofilter, it could be co metabolically degraded in the presence of toluene, benzene, or xylene with the MECs of 34, 23, and 14 g m(-3) h(-1), respectively. The microbial community structure analysis revealed that two large groups could be distinguished based on the presence or absence of MTBE, and many of the dominant bacteria in the consortia were closely related to bacteria isolated from aromatic hydrocarbon contaminated sites and/ or oil wastewaters. These findings provide important new insights into biofiltration and may be used to improve the rational design of biofilters for remediation of petroleum gas-contaminated airstreams according to composition types of mixed gases. PMID- 16719117 TI - Effects of advanced oxidation on green sand properties via iron casting into green sand molds. AB - The effects of advanced oxidation (AO) processing on the properties of green sand were studied via pouring cast iron into green sand molds. Upon cooling, the green sand molds were autopsied at various distances from the metal-sand interface. Autopsy green sand samples collected from a mold that incorporated AO water were characterized and compared to controlled samples collected from a similar autopsied mold made with conventional tap water (TAP). It was found that the AO processing removed a coating of coal pyrolysis products from the clay surface that typically accumulated on the clay surface. As a result, the AO-conditioned green sand retained 10-15% more active clay as measured bythe standard ultrasonic methylene blue titration than did the TAP-conditioned green sand. The AO processing also nearly doubled the generation of activated carbon from the normalized amount of coal composition of the green sand during the casting process. The AO-enhanced activated carbon generation and the AO-incurred clay surface cleaning provided the AO-conditioned green sand with higher normalized pore volume, and thus higher normalized m-xylene adsorption capacity, i.e., relative to before-metal-pouring conditions. Furthermore, mathematical analysis indicated that the AO-conditioned green sand better retained its important properties after pouring than did the TAP-conditioned green sand. Effectively, this meant after metal pouring, the AO-conditioned sample offered about the same net properties as the TAP-conditioned sample, even though the AO-conditioned sample contained less clay and coal before metal pouring. These results conformed to the full-scale foundry empirical finding that when AO is used, foundries need less makeup clay and coal addition through each casting cycle, and they release less air emissions. PMID- 16719118 TI - Interspecies correlation estimates predict protective environmental concentrations. AB - Environmental risk assessments often use multiple single species toxicity test results and species sensitivity distributions (SSDs) to derive a predicted no effect concentration in the environment, typically the 5th percentile of the SSD, termed the HC5. The shape and location of the distribution are best known when populated with numerous toxicity values. To help overcome the cost of multiple toxicity tests, we explored the potential of the U.S. EPA's Interspecies Correlation Estimation (ICE) program to predict single species toxicity values from a single known toxicity value. ICE uses the initial toxicity estimate for one species to produce correlation toxicity values for multiple species, which can be used to develop SSD and HC5. To test this approach to deriving HC5, we generated toxicity values based on measured toxicity values for three surrogate species Pimephales promelas (Fathead minnow), Onchorynchus mykiss (Rainbow trout), and Daphnia magna (water flea). Algal taxa were not used due to the paucity of high quality algal-aquatic invertebrate and algal-fish correlations. The compounds used (dodecyl linear alkylbenzenesulfonate (LAS), nonylphenol, fenvalerate, atrazine, and copper) have multiple measured toxicity values and diverse modes of action and toxicities. Distribution parameters and HC5 values from the measured toxicity values were compared with ICE predicted distributions and HC5 values. While distributional parameters (scale and intercept) differed between measured and predicted distributions, in general, the ICE-based SSDs had HC5 values that were within an order of magnitude of the measured HC5 values. Examination of species placements within the SSDs indicated that the most sensitive species were coldwater species (e.g., salmonids and Gammarus pseudolimnaeus). These results raise the potential of using quantitative structure activity models to estimate HC5s. PMID- 16719121 TI - Comment on "vehicle self-pollution intake fraction: children's exposure to school bus emissions". PMID- 16719119 TI - Effects of the feedlot contaminant 17alpha-trenbolone on reproductive endocrinology of the fathead minnow. AB - Trenbolone acetate is a growth promoter widely used for beef production in the U.S. Two biologically active metabolites of the acetate, 17beta- and 17alpha trenbolone, are ligands of vertebrate androgen receptors and comparatively stable in the waste of treated animals. Both have been detected in surface water associated with beef feedlots, suggesting a potential risk to aquatic animals. In previous work we evaluated the effects of beta-trenbolone on reproductive endocrinology of the fathead minnow (Pimephales promelas) in a 21-day test. The purpose of the present study was to conduct a similar set of experiments with alpha-trenbolone which, based on binding to mammalian androgen receptors, was expected to be less potent than beta-trenbolone. Fecundity of the fish was significantly reduced by alpha-trenbolone with an EC50 (95% confidence interval) of 0.011 (0.007-0.016) microg/L. In females, alpha-trenbolone reduced plasma vitellogenin and steroid concentrations and also induced the production of dorsal nuptial tubercles, structures normally present only in spawning males. Overall, effects of alpha-trenbolone on the reproductive system of the fish were qualitatively and quantitatively quite similar to those caused by beta trenbolone. Part of this similarity might arise from the fact that a substantial amount of the alpha-trenbolone appeared to be converted to beta-trenbolone by the fish. Tissue concentrations of the beta-isomer were consistently similar to or greater than concentrations of alpha-trenbolone, despite the fact that no beta trenbolone was detected in the exposure water. The present study demonstrates the importance of considering both alpha- and beta-trenbolone in assessing the potential ecological risk of androgens associated with beef feedlot discharges. PMID- 16719120 TI - Hypoxia affects sex differentiation and development, leading to a male-dominated population in zebrafish (Danio rerio). AB - Hypoxia is affecting thousands of square kilometers of water and has caused declines in fish populations and major changes in aquatic communities worldwide. For the first time, we report that hypoxia can affect sex differentiation and sex development of zebrafish (Danio rerio), leading to a male-biased population in the F1 generation (74.4% +/- 1.7% males in the hypoxic groups versus 61.9% +/- 1.6% males in the normoxic groups, n = 5; p < 0.05, chi2 test). The increase in males was associated with downregulations of various genes controlling the synthesis of sex hormones (i.e., 3beta-HSD, CYP11A, CYP19A, and CYP19B) as well as an increase in the testosterone/estradiol ratio. The male-dominated populations caused by hypoxia will have reduced reproductive success, thereby threatening the sustainability of natural fish populations. PMID- 16719122 TI - [Insertion of cost teaching in the discipline of administration applied to nursing]. AB - The objective of this text was to make theoretical rescue of the course of the undergraduate discipline Administration Applied to Nursing throughout the history of teaching of Nursing in Brazil, connecting it to the teaching of Costs. We noticed, inserted in the teaching of this discipline, many direct quotes related to the themes of Costs or Economics in Health, from the earliest curriculum in Nursing, dating back to 1890, to the National Curricular Policies of the Nursing Undergraduate Course, of 2001, in addition to indirect quotes related to management, decision taking, and advising in health projects, among others, but with no continuity. PMID- 16719123 TI - [Musculoskeletal diseases, work and lifestyle among public workers at a health institution]. AB - This is a cross-sectional study developed with 651 workers of various occupations. They answered a questionnaire covering general, labor and lifestyle data, in addition to the Work Ability Index, which has been developed by researchers from Finland. In terms of sociodemographics, the analysis of the factors associated with the occurrence of musculoskeletal disorder revealed them to be female, older, and with low level of schooling. As for labor characteristics, the disease was associated with those individuals with physically demanding work, longer working hours on a second job and longer working hours at the institution itself. Regarding lifestyle, the characteristics associated with the occurrence of the disease were obesity, long housework hours and absence of leisure activities. PMID- 16719124 TI - [Nursing diagnosis in the perioperative period of cardiac surgery]. AB - This study identified the nursing diagnoses of patients in the perioperative period of cardiac surgeries and verified the existence or not of agreement between the first author and other nurses in the identification of these diagnoses. Seventeen patients admitted for cardiac surgery were submitted to evaluation using instruments for the perioperative period. In the preoperative, out of the 9 diagnosis categories identified by the first author, three agreed with the nurses (example: Activity intolerance). In the trans-operative, seven categories showed agreement among the nurses and the first author (example: Risk for infection). In the post-operative, 11 categories showed agreement among the nurses and the first author (example: Risk for peripheral neurovascular dysfunction). PMID- 16719125 TI - [Urinary incontinence restrictions in women's life]. AB - Our purpose with this study was to verify the restriction of the urinary incontinence (UI) in a woman's daily life, considering the type of incontinence, and find out how women deal with that. It is a secondary analysis of data taken from a previous study that were obtained using open and closed questions. Interviews were held with 164 incontinent inpatients of gynecological and urological clinics of two hospital schools in the city of Campinas, in the State of Sao Paulo, with ages ranging from 25 to 85 years. Of this universe, 104 (64%) indicated one or more restrictions regarding their daily life activities, such as altered sexual (40.9%), social (33.5%), domestic (18.9%) and occupational activities (15.2%). Mixed urinary incontinence and urge incontinence were mentioned as that affected the most women's daily life. The disposable pad system was the most utilized strategy in dealing with UI. It was concluded that UI has a negative impact on the daily life of these women. PMID- 16719126 TI - [The meaning of being a mother of a child with a heart disease: a phenomenological study]. AB - The objective of this study is to understand what it means to a mother to have a child with a heart disease. In order to do so, a phenomenological approach was used. The study was carried out with mothers of children with congenital cyanotic cardiopathy at the Infant Cardiology Center of a public hospital in Campo Grande, State of Mato Grosso do Sul. Ten open interviews were made based on the following guiding question: What does it mean to you to have a child with a heart disease? The comprehensive analysis of the reported speeches made possible the construction of three categories: "Discovering the disease: being up against the unknown"; "Living together with cardiopathy: being with the child in the disease", and "Believing in a stronger force: the necessary support". The impact of the disease on the mother begins the moment the child is born. Desperation, guilt and insecurity are mentioned as present feelings. It is in God, however, that such mothers look for the necessary support to follow this new path--a path of pain and faith. PMID- 16719127 TI - [Maternal mortality on the family members' perspective]. AB - Maternal mortality is one of the health and social development indicators for countries. It is a tragedy for the family, as the death of a mother deprives the child of breastfeeding and maternal contact and also due to the fact that women maintain family unity. This study is aimed at understanding the meaning of death for the family members of women who have died. The population was formed by 10 family members of 7 women who died as a result of maternal problems in the municipality of Ribeirao Preto. The authors visited their homes and collected data through interviews. For analyzing the data the authors used thematic analysis. Three thematic categories were found: the meaning of maternal mortality; living maternal mortality in the family; and living maternal mortality in the health services. The visit confirmed that there are secondary factors that influenced in the occurrence of maternal deaths. PMID- 16719128 TI - [The effect of immersion baths on the length of childbirth labor]. AB - The objectives of this experimental, randomized, controlled trial study were to evaluate the effect of immersion baths on the length of the first stage of childbirth labor and on the frequency and length of the uterine contractions. Data were collected in a philanthropic public maternity hospital of the city of Sao Paulo whose month average is 1,100 births. The sample was comprised of 108 women in labor--54 in the control group and 54 in the experimental group that had immersion baths. The results showed that immersion baths did not have any influence on the length of labor and on the frequency of uterine contractions. However, the length of contractions was statistically shorter in the experimental group (experimental 41.9 versus control 44.6 min). The conclusion was that immersion baths are an alternative for the woman's comfort during labor, since it provides relief to her without interfering on the labor progression or jeopardizing the baby. PMID- 16719129 TI - [Inspection of the Hospital Infection Control Programs of Health Services by the Health Surveillance: a diagnosis of the situation]. AB - This study was aimed at recognizing and comparing the inspection conditions of the Hospital Infection Control Programs (HICP) through agents of two Technical Groups of Health Surveillance, in 2002, with the use of questionnaires. The results show that the agents have some knowledge about HI, but most of them did not inspect HICPs, were not trained to do so and did not follow specific guidelines nor used other resources. The most mentioned type of inspection was the inspection of physical structure, followed by HICP meeting records and statistics. It was concluded that the main difficulties in HICP inspection are concentrated on the lack of personnel, resources, motivation and technical capacity. It was also observed in the results a heterogeneity between the groups. PMID- 16719130 TI - [Patient satisfaction with nursing care: a comparison between two hospitals]. AB - This is a descriptive study with the objective of measuring and comparing patient satisfaction with the care provided by nurses as well as verifying differences in the satisfaction level in relation to the study's variables in two Sao Paulo hospitals. For data collection was used the Patient Satisfaction Instrument, translated and validated in Brazil. The sample was comprised of 211 patients, of which 128 from Hospital A and 83 from Hospital B. The statistical program SAS was used in the data analysis. The results showed that all patients reported a high level of satisfaction, and the five situations with highest averages were related to the domains Confidence and Professional. However, Hospital A patients showed higher satisfaction level than those of Hospital B. Considering the age variable, only the patients below the age of 60 in Hospital B tend to be more satisfied than the others. PMID- 16719131 TI - [Knowledge and practices of university day care center workers relative to acute respiratory infections in childhood]. AB - Given the increasing frequency of acute respiratory infections in day care centers, the objective of this work was to identify knowledge and practices of day care center workers relative to the prevention, precocious detection and management of these illnesses. Through fourteen structured interviews in two university day care centers of the city of Sao Paulo, it was verified that the workers: demonstrate familiarity with several respiratory illnesses; indicate fever and noisy breath as danger signals; consider pollution and transmission as main causes; refer to environment care as the best form of control; keep continuous observation of the child and interventions of hygiene and comfort, and greater emotional care; consider education and care complementary in child attendance, revealing the need for better preparation for caring; have in daily activities the source of their knowledge. PMID- 16719132 TI - [The influence of anxiety in coping strategies used during the pre-operative period]. AB - This study aims at characterizing the population analyzed regarding socio demographic aspects, identifying the anxiety level of patients undergoing surgery and the most used coping strategies during the pre-operative period, as well as observing the relation between anxiety level and coping strategies over the same period. The population was comprised of 40 patients undergoing surgery. For data collection, two self-applicable instruments were employed: Spielberger's anxiety level inventory and Lazarus, Folkman's coping strategies inventory. Data were collected over the pre-operative period. Most patients were women averaging 46 years old, with low level of schooling, previous surgery experience set before, and no previous intercurrence. The coping strategies more commonly used were social support and problem resolution. In regards to anxiety and coping strategies there was found a negative correlation between anxiety level, social support and problem resolution. PMID- 16719133 TI - [Social organizations and governmental institutions: perspectives on partnerships in children's health care through volunteers and the Pastoral da Crianca]. AB - The aim of this research is to present perspectives on partnerships between social organizations and governmental institutions in children's health care. This study reflects on social participation and relations between governmental and non-governmental services in constructing the consolidation of the Sistema Unico de Saude (Unified Health System), highlighting the role of volunteers and health professionals in this process. In child care, these associations are potential, due to the wide range and prominence of social organizations oriented towards children, particularly the Pastoral da Crianca (the Catholic Church's Child Pastoral), which makes it important to discuss public policies aimed at establishing and strengthening these links in the local and national spheres. PMID- 16719134 TI - [Psychometric contributions to the assessment of psychosocial responses in nursing]. AB - This article presents the methodological bases that guide the development of instruments to measure psychosocial variables. It describes the theoretical, empirical or experimental and analytical and exemplifies the theoretical steps through the experience of developing an instrument to measure powerlessness as a nursing diagnosis. The most difficult procedures for developing instruments to are the theoretical steps. They require the development of theoretical sensitivity as a continuous going forward and backward between concrete and abstract issues; they also require the development of tolerance towards ambiguity, but without abandoning the examination of evidences. PMID- 16719135 TI - [Continuing education in psychiatric nursing: a reflection on concepts]. AB - The concern with continuing education programs that prepare nurses to work in psychiatric nursing led us to conduct this study, the first one on the "development of psychiatric and mental health nursing personnel" research line. Its objective is to make a reflection on the concepts of continuing education, in service and permanent, found in recent nursing education literature. To carry out the literature review of the concepts we reviewed the MEDLINE and LILACS data bases, theses and dissertations and a number of classical titles on the theme. After the analysis of the concepts and discussions with experts, we concluded that the continuing in-service education denomination is the one that presents more consonance with the authors and experts that were consulted, as well as the one that better meets the demands of knowledge in the area of psychiatric nursing towards the quality of assistance. PMID- 16719136 TI - [The nursing diagnosis implementation process at the University Hospital of the University of Sao Paulo]. AB - More than two decades ago the Nursing Department at the University Hospital of the University of Sao Paulo implemented the care model called Nursing Care System (NCS), which is comprised of three phases: Background, Evaluation and Nursing Prescription. Since then, it is being developed by the Nursing Department nurses as a guiding instrument for care, teaching and research. Having in mind the automation of the NCS, the nurses began to discuss the need for changes that could speed up the work process with the proposal of implementation of the Nursing Diagnosis as another step of the NCS and with the review of nursing action/interventions. In order to do so, the adoption of a standardized system of the care process language became essential so as to make possible data capture, grouping and classification for analysis and their transformation into information. The present study aims at sharing with other nurses this experience on the implementation process of the Nursing Diagnosis as a second step of the NCS. PMID- 16719138 TI - [Mental illness in the view of patients and their families]. AB - With the aim of studying the comprehension of mental illness of patients and their families, we conducted a bibliographical study of articles in Brazilian periodicals, theses and dissertations in the period between 1993 and 2003. The identification of the sources was done through the automated search systems Latin American Literature on Health Sciences (LILACS) and the University of Sao Paulo's Bibliographic Data Bank On-line Global Catalog (DEDALUS). Nineteen papers were selected and four categories identified: family difficulty in the relationship with the patient; prejudice and stigma; organic/ biological explanation for the illness; and fear and pain of insanity. Most of the papers show that, regarding assistance, there is a need for support and expansion of the public health system in order to meet the demands. It is believed that the number of publications is small considering the importance of the problem, showing the need for new research. PMID- 16719137 TI - [Leadership in the nursing context]. AB - Leadership is one of the most indispensable tools for nursing work. This study is aimed at making a bibliographical survey about leadership in the nursing context and characterizing the scientific production on the topic based on year and place of publication and theme categorization. An online literature review, carried out using LILACS data, considered periodical material, dissertations and theses available in Portuguese and Spanish from the past twenty years. From the 31 selected works, 17 (54.8%) have been published in international nursing magazines, 10 (32.2%) have been published in Brazilian nursing periodicals, 3 (9.8%) were Master's Degree theses, and one (3.2%) was a teaching thesis, with publication concentrated between 1997 and 1998 (22.6%). When categorizing the references, the numbers were: 14 (45.2%) dealt with nurse leadership styles, 15 (48.5%) were reflections upon leadership, and 2 (6.4%) discussed leadership and communication. PMID- 16719139 TI - [Rehabilitation nursing: expanding horizons, legitimizing knowledge]. AB - The author makes reflections on Rehabilitation Nursing, marking the trajectory of this area in Nursing in terms of care, teaching and research. She discusses concepts and their applicability in Rehabilitation Nursing practice and presents the current demand for care in rehabilitation, as well as the capacity-building of nurses to work in this market, highlighting the pioneering initiatives of public institutions and higher education establishments in the city of Sao Paulo, which has made possible the specialization of nurses in Rehabilitation. PMID- 16719140 TI - [Work and health in the nursing day-to-day routine in a public emergency service]. PMID- 16719141 TI - MSD spring retreat. PMID- 16719142 TI - What physicians can do about the tobacco problem: best practice interventions for your practice and office. AB - This article is the first in a series of four providing up-to-date information about the tobacco problem and how physicians can effectively intervene. Three of the four articles are accredited for free continuing medical education (CME) credit for physicians. The CME credit is being provided free of charge by Nemours via the Web site www.pedseducation.org. Physicians may obtain credit by logging on to this site, registering, and completing the post-test and evaluation materials. This article reviews the best evidence-based approaches for tobacco intervention in the busy office. It discusses the implementation of the following components of a tobacco-control program to effectively and efficiently address the tobacco problem: 1) physician counseling to promote smoking cessation, 2) strategies to promote abstinence from smoking, 3) strategies to minimize environmental tobacco smoke exposure, 4) strategies to encourage tobacco use cessation, and 5) office-based interventions. By reading the article and completing the post-test, it is anticipated that the following learning objectives will be met: 1) physicians will become familiar with best practice interventions for tobacco counseling and limiting secondhand smoke exposure, and 2) physicians will become familiar with office based strategies to promote a strong anti-smoking message. PMID- 16719143 TI - Bone densitometry in pediatric patients. PMID- 16719144 TI - Early screening for nephropathy initiative. PMID- 16719146 TI - [Can positron-emission tomography modify the management of non-small-cell lung cancer?]. PMID- 16719145 TI - AHEC aims to recruit primary care physicians for underserved areas in Delaware. PMID- 16719147 TI - [Does chemotherapy influence the postoperative mortality?]. PMID- 16719149 TI - [Can therapeutic strategy favor the passage to chronicity in metastatic non-small cell lung cancer?]. PMID- 16719150 TI - [Maintenance chemotherapy in advanced NSCLC]. PMID- 16719151 TI - [What is to be expected from population studies in relation to clinical trials?]. PMID- 16719152 TI - [Is small cell lung cancer an orphan disease?]. PMID- 16719148 TI - [Current data concerning neoadjuvant chemotherapy]. PMID- 16719153 TI - [From ASCO and WCLC 2005 to clinical practice: "conventional" treatments]. PMID- 16719154 TI - [From ASCO and WCLC 2005 to the clinical practice: targeted therapies]. PMID- 16719155 TI - [Promising targeted molecular therapies for the future years in non-small-cell lung cancer]. PMID- 16719156 TI - [Megachemotherapy followed by autologous haematopoietic stem cell rescue in children with high risk CNS tumours]. AB - AIM: the Lublin bone marrow transplantation unit experience in megachemotherapy followed by autologus haematopoietic stem cell transplantation in children with high-risk CNS tumours is described. MATERIAL AND METHODS: 9 patients (8 boys and 1 girl) treated in our department between 1999-2004, with high risk malignant brain tumours were included into the study. Median age of the patients was 10 years (range 4-18 years). The group consisted of 5 cases of medulloblastoma, and single cases of ependynmoma, teratoma, astrocytoma anaplasticum and glioblastoma. Stem cell apheresis from peripheral blood was performed during chemotherapy after surgery. Megachemotherapy consisted of carboplatin in total dose 1200 mg/m2, etoposide in total dose 800mg/m2 and melphalan in total dose 200 mg/m2 in five patients. Remaining four children received carboplatin in total dose 1800 mg/m2, etoposide in total dose 750 mg/m and tiothepa in total dose 900 mg/m2. RESULTS: two patients relapsed one and two months after megachemotherapy respectively. No transplant related mortality was observed. Seven children are alive and well with the median observation time 23 months. CONCLUSION: megachemotherapy followed by stem cell rescue is a safe and feasible procedure in children with malignant brain tumours and may improve results of the treatment in high-risk patients, but these results should be con firmed in larger series of patients. PMID- 16719157 TI - [Malignant neoplasms localised in the parameningeal region in children treated at two Polish oncological centres-diagnostic and therapeutic dilemmas]. AB - Malignant neoplasms localized in the parameningeal region include mainly soft tissue sarcomas (MTM), non-Hodgkin s lymphomas (NHL-B) and, less frequently, nasopharyngeal carcinomas. The aim of the study was to analyze diagnostic and therapeutic problems in children with parameniingeal neoplasms treated in Departments of Paediatric Oncology in Gdansk and Lublin between 1992 and 2004. MATERIAL AND METHODS: The study includes 32 patients (M/F: 23/9), aged 2 to 17 years, mean 6,3 years. In 17 children MTM was diagnosed: in nine NHL-B-cell and in six--nasopharyngeal carcinoma (lymphoepithelioma). The diagnosis of NHL-B and undifferentiated MTM were made in two children treated previously for NHL-nonB and retinoblastoma. Two cases of NHL appeared in a girl with ataxia teleangiectasia syndrome. RESULTS: Initial symptoms lasted from 2 weeks to 24 months, mean 4,5 months for the whole group. In NHL patients mean period ofsymptoms was 4,5 weeks, in MTM- 5,5 months and in lynmphoepithelioma--7 months. Symptoms associated with the tumours localisation (snoring, breathing through the mouth, epistaxis, chronic purulent rhinitis, dysphagia and earache) predominated and were treated initially as upper respiratory tract infections. Cervical lymph nodes enlargement was observed in 30% children with MTM and 83% with lymphoepithelioma. Most of patients presented with highly advanced stages of neoplasms. MTM and NHL-B treatment was conducted according to the protocol approved by the Polish Paediatric Solid Tumours and Leukaemia/Lymphoma Studies Group. In patients with lymphoepithelioma different treatment schemes were administered, including chemo- and radiotherapy. Good response to therapy was found in 13/32 patients (41%). The group included 24% children with MTM (all with embryonic subtype), 56% with NHL-B and 67% with lymphoepithelioma. All these patients attained complete remission after standard line I therapy. But 13 children with MTM, four with NHL-B and two with lymphoepithelioma required more aggressive line II treatment because ofpoor response to therapy (NR) or relapse. Finally, 20 of 32 followed-up patients (62,5%) are in durable complete remission from 10 months to 11 years 4 months (mean 4 years) after therapy discontinuation. This group consists of all nine patients with NHL-B, 67% children with lymphoepithelioma and 41% with MTM. In six children (30%) persistent complications of oncological treatment occurred, including: hypoacusia, postradiation defect of the eye ball, postsurgical facial nerve palsy and cranio nasal fistula complicated with pneumocephaly. A patient with MTM of maxillary sinus developed a second neoplasm 2 years after first therapy. This was glioblastoma multiforme located in the left parietal lobe (outside the radiation field). At present, the boy is in complete remission 2,5 years after treatment for the second tumour Among 32 children with parameningeal neoplasms 11 patients died (nine with MTM and two with lymphoepithelioma), all in the phase of disease progression (five NR and six after relapse). In two of them (with MTM) the direct cause of death was myelosupressive, gastrotoxic and infectious complications of antitumour therapy. One child still undergoes treatment for MTM relapse. PMID- 16719158 TI - [Influence of surgery and radiotherapy on growth and pubertal development in children treated for brain tumour]. AB - INTRODUCTION: The increasing number of childhood cancer survivors has resulted in a growing interest in the late effects, which depend on type of treatment. Frequently, a brain tumour and its therapy in children are endocrinologically devastating. AIM OF STUDY: The aim of study was to compare growth and pubertal development in children after brain tumour therapy treated or not treated with recombinant growth hormone (rGH). MATERIAL AND METHODS: 18 children were included in this study. Group I - (12/18) not treated with rGH, after total resection of brain tumour: craniopharyngeoma (8/12), astrocytoma (2/12) ependymoma (1/12), germinoma (1/12). Mean time of remission was 5,0yrs (+/- 0,9). Group II - (6/12) treated with rGH, after subtotal resection of craniopharyngeoma (4/6), ependymoma (1/6), medulloblastoma (1/6) and cranial irradiation with mean total doses 46,5 Gy (+/- 5,65). Children were qualified for rGH replacement according to deceleration of growth and lower growth hormone secretion (< 10 ng/ml) in stimulating tests. Mean time of remission was 6,5 yrs (+/- 2,41). Growth, height in centimeters converted to standard deviation score--SDS, body mass index (BMI), pubertal status and hormonal tests, were also evaluated. RESULTS: All patients were treated with surgery with no cranial irradiation in prepubertal age. 100% children of group I needed substitution because of secondary hypothyreosis, 83% due to secondary adrenal insufficiency and 53% of diabetes insipidus. Mean height after brain tumour surgical treatment in group I was - 1,24 SDS (+/- 0,85) and did not significantly change in the time of observation. Two girls needed hormonal substitution for hypogonadotropic hypogonadism. Mean BMI after total resection of brain tumour was 18,09 (+/- 4,20) and significantly increased to 23,73 (+/- 2,82). In group II - all children presented multihormonal pituitary insufficiency. Mean deviation score of height before rGH treatment was - 3,84 SDS (+/- 2,87) and after mean time of rGH therapy of 1,5 yrs (+/- 1,2) decreased to 2,6 (+/- 1,06). Mean BMI before treatment with rGH 18, 06 (+/- 4,4) increased to 22,41 (+ 0,74) in the time of observation and decreased to 18,5 (+/- 2,87) after 1,5 years (+/- 1,2) of rGH treatment. CONCLUSIONS: 1. Children treated with surgery for brain tumour need substitution for secondary hypothyroidism, part of then need treatment for secondary adrenal and gonadal insufficiency and diabetes incipidus. 2. Children who were treated with surgery and/or cranial irradiation developed multihormonal pituitary insufficiency, growth failure and replacement rGh therapy was needed. 3. Total resection of brain tumour without chemo- and radiotherapy did not impair growth in first years after surgery. PMID- 16719159 TI - [Solid neoplasms localised within the parameningeal region and CNS infiltration]. AB - INTRODUCTION: Solid neoplasms primarily localised within parameningeal region constitute a significant diagnostic and therapeutic problem. Solid tumours developing in the primary site are soft tissue sarcomas (STS) and lymphoepithelioma. AIM OF STUDY: Evaluation of the incidence of CNS infiltration, histological type of tumour and results of treatment. MATERIAL AND METHOD: The study enrolled 14 patients being treated for solid tumours primarily localised within the perimeningeal region, in the Department of Paediatric Haematology and Oncology of the Medical University of Lublin between 1992 and 2004. The age of patients ranged from 2 up to 17 years (mean 8.5): 4 girls and 10 boys. RESULTS: Soft tissue sarcoma was diagnosed in 10/14 children: rhabdomyosarcoma (RMS) embryonale in 8 patients, RMS alveolare in 1, leiomyosarcoma in 1, and lymphoepithelioma in four children. The stage of disease was III degree in all children (without CNS infiltration). Secondary infiltration of CNS was diagnosed in 2/14 children with STS. CONCLUSION: Secondary CNS infiltration was diagnosed in 2/14 patients. Despite of combined treatment, results of treatment are still poor due to local recurrance with CNS infiltration. PMID- 16719160 TI - [Neurologic symptoms in the course of neuroblastoma in children. Own observations]. AB - INTRODUCTION: Neurological origin and multiple localization of neuroblastoma (NB) in children predisposed to the occurrence of neurological deficits. They usually present as symptoms of spinal cord compression, peripheral nerve palsy or central nervous system metastases. The aim of the study was to analyze retrospectively the frequency and characteristics of neurological disorders in patients with NB, treated in the Department of Paediatrics, Oncology, Haematology and Endocrinology, Medical University of Gdansk, Poland, between 1992 and 2004. MATERIAL AND METHODS: 53 children with NB, aged 1 month to 10 years 10 months were included in the study. RESULTS: Neurological symptoms were present in 16 children (30%), all with advanced NB (stages III and IV). In 12 of them neurological deficits predominated in the medical history, contributing to the neoplasm's diagnosis. Most of the patients (10 children) demonstrated symptoms of the lower limbs paresis and bladder/bowel sphincter dysfunction. Four children suffered from severe back pain. Mentioned neurological disorders preceeded NB diagnosis for median period of 2,5 months. In three patients neurological complications of surgical tumour resection occurred (Horner syndrome in two and foot dorsiflexorparesis in one child). No neurological side effects of chemo- and radiotherapy were observed. Neurological symptoms recovered completely with oncological treatment in eight patients, while in three a considerable neurological improvement was observed. No recovery of neurological deficits was obtained in patients with post-surgical complications and in children suffering from disease progression. Three patients died of NB dissemination, 13 children are alive including two patients undergoing therapy. CONCLUSIONS: 1. Most of NB patients recovered completely or partly from neurological disorders while on therapy. Persistent symptoms of neurogenic bladder with accompanying urinary tract infections constitute a considerable clinical problem in some children. 2. NB survivors require intensive interdisciplinary medical care of paediatric oncologist, neurologist, and nephrologist. PMID- 16719161 TI - [Diagnostic and therapeutic difficulties in soft tissue sarcomas localized in nonparameningeal head and neck region--own experiences]. AB - INTRODUCTION: Soft tissue sarcomas (MTM) localized within the nonorbital and non parameningeal head and neck region in children are associated with favourable prognosis. However in our material we have observed many therapeutic failures in this group of patients. The aim of the study was to analyze the reasons for disappointing results of oncological therapy in children with MTM treated between 1992 and 2004. MATERIAL AND METHODS: Nine children (M/F: 6/3; mean age 5,5 years). Five patients were diagnosed with rhabdomyosarcoma, four--with non rhlabdomyosarcoma, including: angiosarcoma, malignant triton tumour, fibrosarcoma and leiomyosarcoma. RESULTS: mean duration of initial symptoms was 7 months. The first sign of the neoplastic disease in all children was tumour. In as many as five of nine patients initially a false histopathological diagnosis was made based on material obtained from aspiration biopsy of the tumour performed in non oncological centres. This resulted in a significant delay of the proper diagnosis of malignant disease ranging from 2 to 15 months (mean 7 months). That is why all our patients presented with highly advanced stages of MTM or even in the phase of disease recurrence. Therapy was conducted according to the schemes: MMT-89, CWS 91, CWS-96 and CWS-2002. Durable complete remission after the first line therapy was obtained in one child only. Six patients developed MTM relapse and two- progression during chemotherapy. Finally five children remain disease-free after treatment termination with follow-up of 1 to 1,5 years. Four of then had microscopically complete delayed resection of the tumour or the relapse. Four patients died of neoplasm recurrence and progression. In three of them the proper diagnosis was delayed significantly and they were diagnosed in the first or even second relapse of the tumour. CONCLUSION: Unfavourable course and treatment results in MTM located in nonorbital and nonparameningeal head and neck region in our patients result from initial wrong histopathological diagnosis and delayed therapy institution. PMID- 16719162 TI - [Soft tissue sarcomas of bladder and prostate in children treated according to the CWS-96 protocol--report of the Polish Paediatric Solid Tumours Study Group]. AB - AIM: analysis of the course, outcome and therapy complications in bladder/prostate soft tissue sarcomas (STS) in children treated from I'1997 to II'2003 according to CWS-96 protocol in Poland. MATERIAL AND METHODS: 22 children (M/F: 17/5, age: 8 months - 17 years 2 months; median 5,3y). Histopathology: RMS 20 patients (RME-14), non-RMS-2 patients. RESULTS: 96% presented with advanced neoplasm (III - 14, IV - 7). In 18 patients (82%) primary tumour exceeded 5cm; in 17 (77%) extended beyond site of origin. Lymph nodes metastases were stated in six (27%), distant metastases in seven patients (32%). No patient underwent primary complete tumour excision. All received chemotherapy for high risk group. Response to chemotherapy after three cycles was favourable in eight and unfavourable in eight children. Delayed resections were performed in 11 patients (mutilating - 7) proving complete in only five. Radiotherapy (32-50,4Gy) was given to 16 patients, mainly after delayed incomplete surgery. Five patients developed local relapse, four--continual disease progression. None of these nine patients had ever complete tumour resection. Nine patients died: seven due to neoplasm progression, two of therapy complications (septic shock, ARDS). Thirteen patients are alive (59%) with mean follow-up of 42 months. Eight children had bladder removed with continent ileal pouch formed in four. CONCLUSIONS: 1) Advanced stages of bladder/prostate sarcomas in children suggest the necessity of earlier diagnosis. 2) Due to adjuvant chemo- and radiotherapy approximately 60% of patients with this diagnosis may be nowadays cured of bladder/prostate sarcomas without mutilating surgical procedures. PMID- 16719163 TI - [Genitourinary soft tissue sarcomas located outside bladder and prostate in children treated according to the CWS-96 protocol--report from the Polish Paediatric Solid Tumours Study Group]. AB - AIM: Analysis of therapy efficacy in non-bladder/prostate genitourinary sarcomas in children treated from I'1997 to VI'2003 with CWS-96 protocol in Poland. MATERIAL AND METHODS: 19 children (M/F: 15/4, age: 3m-17,5y; median 7,2y). Histopathology: RMS - 15pts (RME-13), non-RMS- 4. RESULTS: Primary site: testes - 9 patients, paratesticular region - 6, uterus - 2, vagina and ovary-1 of each. 63% presented with low stage neoplasm (I - 7, II - 5). Primary tumour exceeded 5cm and/or invaded surrounding tissues in 7 patients (37%). 3 patients had regional, 2 patients--distant lymph nodes metastases. Primary excision: complete in 7 patients, incomplete - 12 (microscopically - 5, macroscopically - 7). Six of 7 patients with macroscopic tumour residues responded to chemotherapy (CR-4, GR 2). One patient (stage III triton tumour of uterus) did not, respond but obtained complete remission after mutilating delayed surgery. No other patient required delayed tumour resection. Radiotherapy (23,5-54 Gy) was given to 8 patients. 3 children developed local relapse, 3 patients died (16%): 2 due to neoplasm progression, 1 of neutropenia-related sepsis. 16 patients are alive (84%) with mean follow-up 48 months. The only permanent complications result from mutilating surgery. CONCLUSIONS: 1) prognosis in children with non-bladder/prostate genitourinary sarcomas is favourable despite incomplete primary excision of the neoplasm. 2) chemotherapy and radiotherapy were accompanied by severe but transient myelosupression in the HR group. PMID- 16719164 TI - [Late side-effects of treatment in patients with nephroblastoma]. AB - The aim of the study was to analyse late side-effects of the antineoplastic treatment in patients with diagnosed nephroblastoma. MATERIAL AND METHODS: In the years 1986-2002, 80 children were treated in the Gdansk Centre, for nephroblastoma, 13 of them followed SIOP-6, 25 were treated according to SIOP-9 and the remaining 42, SIOP 93-01 programme. RESULTS: 70 patients had long-term clinical remission. 10 patients died, 3 of them due to treatment complications (all were treated in accordance with SIOP 92-01: 1 - mycotic sepsis, 1 intra operative hemorrhage and 1 - typhlitis), the other 7 due to disease progression (2 on the SIOP-6, 2 on the SIOP-9 and 3 on the SIOP 93-01 protocol). The late diagnosed complications of the treatment were: cardiomiopathy in 7 children, renal insufficiency in 1 case (the girl is presently after kidney transplant), tubulopathy in 4, proteinuria in 3, arterial 1 hypertension in 6, scoliosis in 9, chronic hepatitis (exclusively in children treated before 1994): type B in 11 (one girl has portal hypertension and esophageal varices), type C in 12 and both B and C in 5 children. Of the endocrinologic complications 4 patients were diagnosed with hipergonadotrophic hypogonadism. Secondary, neoplasms that were diagnosed several years from the completion of the nephroblastoma treatment, were the cause of death in 3 children (2 osteosarconma and 1 chondrosarconma). CONCLUSIONS: The advance in the treatment of neoplasia that has been achieved due to modern diagnostic and therapeutic procedures and the continual increase in the number of survivals obliges us the provide the patients with extensive specialistic and continuous medical care. PMID- 16719165 TI - [Relapses in children with nephroblastoma--a single centre experience]. AB - The aim of the study was to assess the incidence of relapse and outcome of treatment in children with nephroblastoma relapse. MATERIAL AND METHODS: In the years 1989-2002 at our centre 67 children were treated for nephroblastoma. Patients were divided into two subgroups. Group I comprised 25 children treated in accordance with the SIOP-9 programme, whilst group II consisted of 42 patients following the SIOP 93-01 protocol. RESULTS: In group I, five children had relapse diagnosed: local in 2, pulmonary in 2 and cerebral in one case. Two of the cases were diagnosed during the treatment, and the other 3 after 1 to 19 months from the completion of treatment. In group II relapse was diagnosed in 4 children: local in 2 and pulmonary in 2 cases. One case was diagnosed while on treatment, and the other 3 were diagnosed 1,2 and 4 months after completion of treatment. Among the children who followed the SIOP-9 programme three patients died, while among those following SIOP 93-01 two patients died. CONCLUSIONS: The prognosis in patients who relapsed remains uncertain. The histological type of tumour and the moment of relapse occurrence are significant prognostic factors. PMID- 16719166 TI - [Mediastinal involvement in the course of proliferative diseases in children in the materials of one institution]. AB - The mediastinum region may be the primary or secondary localization of neoplasms. The aim of our study was the assessment of clinical symptoms, histopathology and outcome of mediastinal tumours in children. MATERIALS AND METHODS: A group of 27 children, chosen from the group of 128 patients with malignancies, which were treated in the years 2000-2004 in the Oncology and Haematology Unit of the Paediatric Department in Katowice, has been studied. In this group there were 14 boys and 13 girls at the age between 2 and 12 years. We analysed the clinical symptoms prior to the diagnosis and the duration of these symptoms, histologic type of tumours and results of treatment. RESULTS: The primary localization in the mediastinum was diagnosed in 23 patients (85,2%) among them 15 (55,5%) of these children have been diagnosed as having Hodgkin's disease, and in 8 (29,6%) non Hodgkin's lymphoproliferative disease. Four of the children (14,8%) had mediastinal secondary localization of solid tumours (2 - neuroblastoma, 1 - carcinoma suprarenalis, 1 - carcinoma epitheliale of unknown origin). The most frequent symptoms were: fever (70,3%), weakness (66,6%), cough (55,5%), madiastinalpain (33,3%). The duration of these symptoms prior to the diagnosis was between 5 days and 182 clays. CONCLUSIONS: 1. In the group of neoplasms localized in the mediastinum, lymphoproliferative diseases are the most frequently diagnosed disorders. 2. In case of fever of unclear etiology and persistent cough, it is necessary to exclude mediastinal malignancy. 3. There is a need to improve the system of oncological education for medical students and doctors. PMID- 16719167 TI - [Assessment of treatment results in children with malignant liver tumours in own experience]. AB - AIM: The assessment of malignant liver tumours in children treated in our centre between years 1985-2004 has been made in order to analyze the prognostic factors and improvement in survival rate. MATERIAL AND METHODS: 17 patients with malignant liver tumours were followed-up. There were 10 (58,8%) patients with hepatoblastomas, 5 (29,4%) hepatocarcinomas, 1 (5,9%) undifferentiated embryonal sarcoma and 1 (5,9%) rhabdomyosarcoma. Primary metastatic disease was recognized in 3 cases as: hepatic vascular involvement, lungs, femoral bone and lymph nodes of liver hilus metastases. All patients underwent preoperative chemotherapy. Tumour resection was attempted in 13 (76,5%) cases; it was complete within adequate resection margins in 11 (64 7%). In 3 cases biliary fistulas occurred after surgery. Secondary metastases appeared in lungs, lymph nodes of liver hilus and central nervous system in 4 cases. RESULTS: Twelve patients are alive with median follow-up 34,0 mths, five died with median survival time 16,0 mths. Total excision of liver tumour had no statistical significance in lifetime prolongation (p =0,12). Survival rate was statistically longer in patients without metastatic disease (p=0,028). CONCLUSIONS: Complete surgical excision had no statistical significance in increasing survival time in liver tumour patients. Metastatic disease had statistical significance in shortening overall survival of patients with liver tumours. Unsatisfactory results in hepatocarcinoma treatment in children dramatically demonstrate the need for new treatment approaches. PMID- 16719168 TI - [Cardio-vascular disorders at the time of diagnosis of malignant solid tumours in children--own experiences]. AB - Cardio-vascular abnormalities in children with neoplastic disease are observed in some cases even during initial diagnostic examinations. The aim of the study was to evaluate the frequency and characteristics of cardiological and vascular disorders in the initial phase of malignant solid tumors in children. MATERIAL AND METHODS: The study included 236 paediatric patients diagnosed with malignant solid tumours, treated in the Department of Paediatrics, Haematology, Oncology and Endocrinology, Medical University of Gdank, between 1992 and 2004. Disorders of the myocardium and pericardium as well as abnormalities in vein blood flow were identified. Diagnostic examinations comprised. electrocardiography, echocardiography, chest X-ray, computed tomography and magnetic resonance imaging. Angiography was performed in selected cases. RESULTS: Abnormalities of the heart and vessels were detected in 35 children (15%). Most of them (83%) were associated, directly or indirectly, with neoplastic growth. High frequency of pericardial effusion was detected (16 patients). Cardiologic emergency requiring immediate cardiosurgical intervention,occurred in two children with neoplasms infiltrating myocardium and/or pericardium. Abnormal vein circulation presented as vena cava superior and vena cava inferior syndromes in ten children. Cardio vascular disorders not associated with neoplastic disease (congenital heart and vessel defects, pancarditis) were observed as rare conditions (2,5%) but in most cases resulted in delay in starting chemotherapy and modification of the treatment scheme. Cardio-vascular abnormalities associated with neoplastic growth tended to disappear during active chemo- and radiotherapy. CONCLUSIONS: The results of the study confirm that cardio-vascular abnormalities in children with cancer before therapy institution, are not infrequent. They may be life threatening and influence the course and outcome of therapy. These observations imply the necessity of cooperation between the paediatric oncologist, cardiologist and radiologist during the initial diagnostics of malignant solid tumours in children. PMID- 16719169 TI - [Orbital pseudotumour imitating a proliferative process]. AB - INTRODUCTION: Orbital pseudotumour is a non-specific inflammatory process of the orbit of unknown origin. It is a rarely diagnosed disease particularly in children, which imitates a neoplastic process. Typical clinical picture is a tumour localized in the orbit, causing various degree of exophtalmus and a decrease of globe mobility and vision. The extent of intraorbital changes are revealed by imaging studies (USG, TK, MR). Diagnosis is based on histopathology of tumour sample. In treatment steroid therapy, radiotherapy or chemotherapy in resistant cases are used. Relapse and malignant transformation are observed. CASE REPORT: We present a case of a 5-years old girl with orbital pseudotumour. In the histopathological examination there a small lymphoid cells, immunohistochemically there is mixed lymphocytic T and B infiltration (CD 3 (+), CD 20 (+), bcl (+), CD 43 (-)). She was treated with steroid therapy, and achieved complete regression of the tumour. At present she is regularly oncologically examined because of the possibility of malignant transformation. CONCLUSIONS: 1. Orbital pseudotumour should be included in the differential diagnosis of children with an orbital tumour 2. Corticosteroids seem to be the treatment of choice in orbital pseudotumour 3. Children with orbital pseudotumour should be regularly oncologically examined because of the possibility of malignant transformation. PMID- 16719170 TI - [Prognosis of clinical course of primary brain tumours in children in relationship to the duration and characteristic features of initial clinical symptoms]. AB - Primary tumours of the central nervous system belong to the most frequently occurring neoplastic diseases in paediatric patients. During the initial phase of disease development, the clinical symptoms of brain tumours might suggest disorders of other organs and their diagnosis is frequently delayed in relation to therapeutic possibilities. The aim of the study was to analyse the characteristic features and duration of preliminary symptoms arising due to brain tumours in paediatric patients treated in a single centre and to try to assess their prognostic significance for recurrence and death. MATERIAL AND METHODS: We performed a retrospective assessment of the characteristic features of preliminary symptoms of brain tumours in 81 paediatric patients (45 female, 35 male) in the age range of 1.5 month - 17.2 years. Those characteristic features included the duration of symptoms until diagnosis (Pre-Diagnostic Symptoms Interval, PSI) and their correlation with the tumour type, its localization and size at the moment of diagnosis. RESULTS: The mean duration of symptoms in the studied group was 3 months. In nearly 45% of patients PSI was longer then 3 months and in 17,5% it was over 6 months. The predominant preliminary disease symptoms were the symptoms of increased intracranial pressure (n=56, 69,1%) The longer PSI correlated with the disease recurrence rate (p=0.024) and death rate (p=0.04). When PSI was longer then 6 months, all the tumours diagnosed were larger then 30 mnm, however no relationship was found between PSI duration and the tumor size (p=0.35). There was no correlation between the tumour size and the frequency of death (p=0.8), but in patients with tumours smaller then 30 mm in their greatest dimension, the recurrence of the neoplastic process was more frequent. CONCLUSIONS: Duration of preliminary symptoms may have an effect on the tumour recurrence and on the rate of death in paediatric patients with brain tumours. Early diagnosis plays an evident role in prognosis. PMID- 16719171 TI - [Increased urinary excretion of the neurotoxic side-chain metabolites of ifosphamide in children with polymorphism of the glutathione S-transferases genes]. AB - From 5% to 30% of children treated with ifosphamide (IF) develop symptoms of neurotoxicity due to toxic metabolites of the drug: 2- and 3- dechloroifosphamide (2- and 3-DCIF) and chloracetaldehyde (CAA), which cause glutathione depletion in cells. The aim of the study is to establish the influence of polymorphism of genes encoding for glutathione S-transferases classes pi (GSTP1), mi (GSTM1) and theta (GSTT1) on frequency of neurotoxicity of IF and amounts of toxic metabolites of the drug excreted in urine. MATERIAL AND METHODS: Neurotoxicity of IF was assessed in 76 children (38 girls and 38 boys), aged 9 to 210 months with diffrent kinds of neoplasms. They were treated with IF in 3-hours infusion in doses from 1.5 g/m2 to 3 g/m2 for 3 to 5 days. Before chemotherapy, deletions of GSTT1, GSTM1 genes and transition at +313 A-G in GSTP1 gene were identified with PCR and PCR-FRLP method, respectively. Daily urine excretion of 2-DCIF, 3-DCIF and unmetabolised IF was assessed with nuclear magnetic resonance (31P NMR). RESULTS: Symptoms of neurotoxicity were observed in 14 (18%) of 76 examined children treated with IF Comparing to children without neurological symptoms, in children with encephalopathy urinary excretion of unchanged ifosphamide was lower (p=0.055) and 2DCIF and 3DCIF was increased. Concomitantly, in children with transition at 313 A-->G GSTP1 gene concentrations of 2DCIF and 3DCIF were increased. Excretion of unmetabolised IF was statistically significantly higher in children with deletion of GSTT1 gene (p=0.02). Moreover, no correlation was found between the GSTM1 genotype and the excretion of ifosphamide and its metabolites. CONCLUSION: The results suggest that ifosphamide can be the substrate for glutathione S-transferases. Polymorphism of genes coding for glutathione S-transferases can influence individual reactions to iphosphamide. PMID- 16719172 TI - [To cure a child with cancer. The principles of bio-psychosocial support]. AB - Currently most children with cancer can be cured due to tremendous progress in the treatment methods. A child truly cured is only the patient who is a long-time survivor from a medical point of view as well as with fill psychosocial adjustment. This is the reason for many efforts which are undertaken to establish the principles of psychosocial support of children with neoplastic disease. Also in Poland such activities have started. This multicentre study was performed on the optimalisation of biomedical and psychosocial support of cancer children and their parents based on the recommendations of SIOP Working Committee on Psychosocial Issues in Paediatric Oncology adjusted to Polish conditions. The analysis showed the important role of introducing the principles of biomedical and psychosocial support. The implemented unified model of giving information about cancer to the children tells us who of them needs the additional psychosocial interventions. There is also a neccessity to improve the communication skills of the paediatricians. PMID- 16719173 TI - Palliative care: helping patients live longer--and better--with cancer. PMID- 16719174 TI - Palliative care nurses: the Steel Magnolias of nursing. PMID- 16719179 TI - Should oncology nurses provide spiritual care? PMID- 16719180 TI - Pecking orders, social permits, & reverse discrimination. Interview by Shirley A Smoyak. AB - The first step in moving toward changing the current system is recognizing how these unspoken beliefs and "word compartments" create incredible pain and cause unnecessary delay in providing help. Anne also had eloquent things to say about the failure of parity, as far as people with mental illnesses are concerned. Using an e-mail list, Anne collected reflections of some Vermonters involved in the mental health field. Here are two of my favorites about the term "behavioral health": From a consumer: "To me, it implies if I would just change my behaviors, I wouldn't have a mental illness." From a nurse: "Call it what it is. We have cardiac care units, not happy heart units." If you have your own reflections or experiences with the descriptions in this editorial, do send them to us at jpn@slackinc.com. PMID- 16719181 TI - Considering the dominant discourse. PMID- 16719182 TI - Challenges in the diagnosis & treatment of bipolar depression--part 2: treatment options. PMID- 16719183 TI - Aging workers in search of employment & health insurance coverage. AB - Aging workers, ages 55 to 64, face challenges finding employment and health insurance coverage. In the search for employment, they frequently discover that their skills and needs do not match the opportunities in today's work environment. Roadblocks in the search have the potential to negatively affect overall health and well-being. This article examines employment and health insurance challenges faced by aging workers and suggests nursing interventions that will help reduce stress and promote healthy aging. PMID- 16719184 TI - Nursing in tele-mental health. AB - This article discusses a cooperative project involving an academic health sciences center, a state university, a women's shelter, and a community mental health center in East Texas, a medically underserved, rural region of Texas. The U.S. Department of Commerce provided grant funding to develop a telehealth network to provide an evaluation, referral, and treatment program for victims of domestic violence. Nurses were involved in all phases of the project, from initial conception through development, implementation, and evaluation. The authors concluded that all of the women involved in the study had significant mental health issues and other health concerns that were not being addressed by the existing health care delivery system. Without the use of telehealth, these women would have had limited access to primary health care and virtually no access to mental health services. The East Texas Tele-Mental Health Network demonstrates the value of the clinical and organizational skills that nurses bring to innovative models of mental health service delivery. PMID- 16719185 TI - Interviewing techniques for the Asian-American population. AB - The cultures of racial and ethnic minorities influence many aspects of mental illness, including communication styles, symptoms, coping strategies, family and community support, and willingness to seek treatment. This article presents the effects of Asian American/Pacific Islanders' beliefs and behaviors related to mental health. Strategies to enhance the process and outcomes of the psychiatric interview with members of this populatior are addressed. PMID- 16719186 TI - Educational needs of inpatients with severe & persistent mental illness: a partial replication. AB - Although a great deal has been written about the importance of patient education, few replications of past research have been conducted to increase clinicians' confidence in the conclusions of previous studies. This is especially true of studies conducted in clinical settings where findings may or may not be generalizable to other institutions. This partial replication study tested whether previous findings about patient education in an outpatient setting were applicable to an inpatient setting using a modified version of the instrument. A 1998 study published in this journal suggested that differences may exist between the perceived health education needs of outpatients and their health care providers. In this partial replication, we examined the differences between the educational needs of inpatients and those perceived by their attending nurses. As in the original study, we found discrepancies between patients' actual needs and nurses' perceptions of those needs, supporting the importance of assessing patients' health educational needs in the treatment planning process. PMID- 16719187 TI - Adrenal insufficiency in the critically III trauma population. AB - Acute adrenal insufficiency has been demonstrated in a number of patients with shock. This study was designed to evaluate the rate of occult adrenal insufficiency in the critically ill trauma population and to determine the impact of hypoproteinemia on the use of random cortisol levels as a marker for adrenal insufficiency. Forty-four patients were prospectively enrolled on admission to the trauma intensive care unit, with three excluded, for a total n of 41. Random total serum cortisol and albumin levels were drawn on hospital Days 1, 4, 8, and 14. Occult adrenal insufficiency was defined as a cortisol less than 25 mcg/dL in the setting of an albumin greater than 2.5 g/dL. The prevalence of cortisol less than 25 mcg/dL ranged from 51 to 81 per cent during the study period, and peaked on Days 4 and 8. Albumin 2.5 g/dL or less ranged from 37 to 60 per cent, and this prevalence also peaked on Days 4 and 8. The patients with a low albumin had a high prevalence of low cortisol, ranging from 67 to 100 per cent. The prevalence of adrenal insufficiency, with low cortisol and normal albumin, ranged from 41 to 82 per cent during the study period. None of our patients with occult adrenal insufficiency were treated with steroids, which was a decision made by the treating physicians. Among the patients with occult adrenal insufficiency, survival was 100 per cent. Occult adrenal insufficiency is common in critically ill trauma patients, and is a dynamic entity that can be acquired and even resolved during critical illness. Random cortisol of 25 mcg/dL may actually not be an adequate marker of occult adrenal insufficiency. Low albumin predicts a low cortisol. Hemodynamically stable occult adrenal insufficiency should not be treated with steroid replacement in the critically ill trauma patient, as survival in our series was 100 per cent without replacement. PMID- 16719188 TI - Complications of brain death: frequency and impact on organ retrieval. AB - Brain death is associated with complex hemodynamic, endocrine, and metabolic dysfunction that can lead to major complications with the potential donor. Untreated, this can progress to cardiovascular collapse with loss of valuable organs for transplantation. We hypothesized that brain death-related complications would have no effect on the number of organs donated if an aggressive donor management protocol was in place. We identified all successful organ donations between January 2000 and December 2003 and evaluated them for brain death-associated complications (defined as vasopressor requirement, coagulopathy, diabetes insipidus, cardiac ischemia, lactic acidosis, renal failure, and acute respiratory distress syndrome) and donated organs per donor. Sixty-nine organ donors were identified. Complications identified were as follows: intravenous vasopressor requirement in 97.1 per cent, coagulopathy in 55.1 per cent, thrombocytopenia in 53.6 per cent, diabetes insipidus in 46.4 per cent, cardiac ischemia in 30.4 per cent, lactic acidosis in 24.6 per cent, renal failure in 20.3 per cent, and acute respiratory distress syndrome in 13 per cent. There was no significant effect of complications on the average number of organs harvested, with the exception of an increase in organs harvested in the presence of diabetes insipidus. With the implementation of an aggressive organ donor management protocol, these complications can be effectively managed with no impact on the number of organs harvested for transplant. PMID- 16719189 TI - Recurrence after complete cryoablation of colorectal liver metastases: analysis of prognostic features. AB - This study critically evaluated the prognostic determinants for disease-free survival (DFS) after cryoablation for colorectal liver metastases. An observational cohort study of prospectively collected data on 135 patients who underwent cryoablation with or without resection for colorectal liver metastases was performed. Univariate and multivariate analyses were used to determine the prognostic factors for overall DFS, cryosite DFS, remaining liver DFS, and extrahepatic DFS. Overall, 115 patients (85%) developed recurrence at the cryosite (44%), and the remaining patients developed recurrence at the liver (62%) and extrahepatic site (71%). In univariate analysis, preoperative and postoperative carcinoembryonic antigen (CEA) were significant for overall DFS. Distribution of metastases, operation type, total number of metastases, number of cryotreated metastases, largest size of cryotreated metastasis, and postoperative CEA were significant for cryosite DFS. The number of cryotreated metastases and postoperative CEA were significant for remaining liver DFS. The largest size of cryotreated metastasis, and preoperative and postoperative CEA were significant for extrahepatic DFS. In multivariate analysis, resection plus cryoablation, < or =7 liver metastases and < or =3 cm cryotreated metastasis were independently associated with an improved cryosite DFS. Preoperative CEA of < or =5 ng/mL was independently associated with an improved overall and extrahepatic DFS. The role of CEA in colorectal metastasis is important. Resection plus cryoablation rather than cryoablation alone should be used for larger lesions. PMID- 16719190 TI - Hepatic tumor ablation: cryo versus radiofrequency, which is better? PMID- 16719191 TI - Prospective evaluation of an extubation protocol in a trauma intensive care unit population. AB - Little data exists regarding extubation protocols in critically injured trauma patients. The objective of the current study was to prospectively examine the impact of implementing an extubation protocol on the outcomes of ventilated trauma patients in a surgical intensive care unit (STICU). Trauma patients admitted to the STICU over a 15-month period at a Level 1 trauma center were prospectively evaluated. The total period was divided into an education and institution period (April 2002-November 2003) and an evaluation period (December 2003-July 2003). Patient demographics, hospital course, complications, and outcomes from period I were compared with those obtained during period II. From April 8, 2002 through July 5, 2003, 69 patients intubated for greater than 24 hours were included in our analysis. Thirty-three were treated during period I and 36 were treated during period II. Both groups were well matched in terms of age, sex, Injury Severity Score, and chest Abbreviated Injury Score. Ventilation days significantly decreased from a mean of 16.3 to 8.2 days (P = 0.04). ICU length of stay also decreased, nearly meeting significance. A rigorously enforced extubation protocol significantly decreased ventilator days in STICU patients. Continued education of health care providers is key to the success of the protocol. PMID- 16719192 TI - Sentinel lymph node biopsy for occult breast cancer detected during breast reduction surgery. AB - Breast reduction surgery is considered a relative contraindication to a sentinel node biopsy because of the possibility that lymphatics have been interrupted by the procedure. We describe six patients who underwent successful sentinel lymph node biopsy for occult carcinomas detected after breast reduction surgery. A subsequent skin-sparing mastectomy, along with a sentinel lymph node biopsy, was performed. Reconstruction was possible in five of six patients. Sentinel lymph node biopsy should not be considered a contraindication after breast reduction surgery. PMID- 16719193 TI - Nonoperative management of spontaneous splenic rupture in infectious mononucleosis: the role for emerging diagnostic and treatment modalities. AB - Infectious mononucleosis (IM) is a self-limiting lymphoproliferative disorder affecting teenagers and young adults. Splenomegaly is a common manifestation of IM and results in a compromised organ that may rarely rupture spontaneously, with significant morbidity and mortality. The IM spleen should be protected from even minor trauma. Although traditional management of spontaneous splenic rupture in IM has been splenectomy, the role of nonoperative management is evolving. The advent of endovascular interventional modalities has augmented the physician's armamentarium in managing these patients nonoperatively. We report a case of spontaneous splenic rupture in a patient with IM managed conservatively with the aid of splenic angiography. The option of arteriography, with or without embolization, should be considered in the management of all patients with spontaneous splenic rupture in the setting of IM. PMID- 16719194 TI - Giant hepatic hydatid cyst as a cause of small bowel obstruction. AB - Today, giant hydatid cysts are fairly rare even in endemic areas. We describe a case of an extremely large hydatid cyst of the liver that was causing massive compression of the neighboring organs, giving rise to a subocclusive syndrome of the intestine. After considering the various approaches available for the treatment of this disease, it was decided that radical surgery was still the best therapeutic choice, especially when extremely large cysts are involved. In particular, in clinical situations where it is difficult to perform radical surgical procedures, less aggressive surgery followed by drug treatment should be the treatment of choice. PMID- 16719195 TI - Acute appendicitis: is there a difference between children and adults? AB - Historically, the lack of classic symptoms and delay in presentation make diagnosing acute appendicitis more difficult in children, resulting in a higher perforation rate. Despite this, the morbidity of acute appendicitis is usually lower in children. We evaluated the current differences in clinical presentation, diagnostic clues, and the outcomes of acute appendicitis between the two age groups. A retrospective review of 210 consecutive cases of pediatric appendectomy and 744 adult cases for suspected acute appendicitis from January 1995 to December 2000. Pediatric patients were defined as being 13 years and younger. Pediatric patients were similar to adult patients with respect to duration of pain before presentation (2.4 +/- 4.3 days vs 2.5 +/- 7.3 days), number of patients previously evaluated (22.0 vs 17.7%), number of imaging tests (computed tomography or ultrasound; 32.9 vs 40.2%), and number of patients observed (16.7 vs 17.2%). However, pediatric patients required less time for emergency room evaluation (4.0 +/- 2.7 hours vs 5.7 +/- 4.9 hours, P = 0.0001). In children and adults, a history of classic, migrating pain had the highest positive predictive value (94.2 vs 89.6%), followed by a white blood cell count > or =12 x 109/L (91.5 vs 84.3%). The overall negative appendectomy rate was 10.0 per cent for children and 19.0 per cent for adults (P = 0.003); the perforation rate was 19.0 per cent and 13.8 per cent, respectively (P > 0.05). The perforation rate in children was not associated with a delay in presentation (perforated cases, 2.9 +/- 3.3 days compared with nonperforated cases, 2.3 +/- 4.6 days). Mortality and morbidity, including wound infection rate and intra-abdominal abscess rate, were similar. Contrary to traditional teaching, diagnosing acute appendicitis in children is similar to that in adults. A history of migratory pain together with physical findings and leukocytosis remain accurate diagnostic clues for children and adults. Perforation rate and morbidity in children is similar to those in adults. The outcomes of acute appendicitis in children are not associated with a delay in presentation or delay in diagnosis. PMID- 16719196 TI - A prospective, randomized trial of cognitive intervention for postoperative pain. AB - A single-blind, randomized prospective trial was performed at a university hospital to determine if preoperative relaxation training will decrease pain and narcotic demand postoperatively. A convenience sample of 49 patients undergoing lumbar and cervical spine surgery was randomized to receive instruction on relaxation techniques or routine preoperative information before surgery. Pain score and narcotic demand in the first 48 hours after surgery were the primary outcomes. Pain scores were higher in the relaxation (4.8 +/- 1.7) versus the standard preparation group (3.9 +/- 1.7) on postoperative day one (POD) 1, but lower on POD 2 (3.9 +/- 1.9 vs 4.1 +/- 1.9), whereas narcotic use (milligrams of IV morphine per hour) was higher in the relaxation group on POD 1 (1.14 +/- 0.94 vs 0.54 +/- 0.55) and POD 2 (0.86 +/- 0.73 vs 0.50 +/- 0.61). The differences were significant for narcotic demand (P = 0.01) but not for pain (P = 0.94). In conclusion, our results could not support the use of relaxation training for reducing postoperative pain and narcotic demand in this selected surgical population. PMID- 16719197 TI - Correlation between clinical and histological findings in parathyroid tumors suspicious for carcinoma. AB - Carcinoma of the parathyroid is a rare malignancy that can be cured surgically if the proper diagnosis and treatment is given initially. Arriving to the clinical suspicion of a malignancy preoperatively is by far the most important step for a good prognosis. Our goal is to review the correlation between clinical and final histopathological findings that can arouse the suspicion of such malignancy and their true predictive value in the diagnosis. All patients that underwent surgical removal of the parathyroid mass between March of 1992 and March of 2003 were reviewed retrospectively at Providence Hospital and Medical Centers. Among 168 patients who underwent parathyroid excision, 14 (8.3%) had hyperplasia of the parathyroid, 121 (72%) had benign adenoma, 25 (14.8%) had other benign lesions, and 8 (4.7%) patients had primary carcinoma of the parathyroid confirmed by pathology. Our mean serum calcium level was 11.57 mg/dL, which was lower than the mean level (12 mg/dL) for benign hyperparathyroidism. The mean tumor size was 2.18 cm, smaller than the proposed for malignant criteria, and none of the eight patients (0%) had any symptoms of hypercalcemia at the time of diagnosis. Seven of eight patients (87.5%) had frank signs of invasion together with other histological features, and two patients had associated papillary carcinoma of the thyroid. Five patients from our series did not meet clinical criteria for malignancy (tumor size > 3 cm, palpable mass, and serum calcium > 14 mg/dL), but had undisputable histological findings (high mitotic pattern, fibrous trabeculae, capsular invasion, vascular invasion, and nodular involvement). On the other hand, 17 patients with benign histology had tumors greater than 3 cm, and an additional 18 had palpable masses on physical examination. We believe that these patients need to be followed closely. The patients with diagnosis of parathyroid carcinoma, their kindred, and those with large adenomas may benefit from genetic screening for HRTP2 gene mutations in search of early detection of tumors suspicious for malignancy. This is based on the fact that we did not find correlation between the clinical presentation and the histological features in our patients with proven malignancy. PMID- 16719198 TI - Giant mesenteric fibromatosis presenting as small bowel obstruction. AB - Mesenteric fibromatosis is a proliferative fibroblastic neoplasia of the small intestine mesentery that may occur as a unique or multiple formation. Mesenteric fibromatosis is a rare, locally aggressive neoplasm and may present with abdominal discomfort, abdominal pain, weight loss, or symptoms of ureteral obstruction, mesenteric ischemia, or intestinal obstruction. It is of the utmost importance to distinguish mesenteric fibromatosis from gastrointestinal stromal tumors. Histopathology accurately differentiates between these two distinct entities. The preferred treatment is local surgical excision with a margin of uninvolved tissue. The involvement of important structures like the superior mesenteric artery and the superior mesenteric vein may pose a challenge during resection, but these tumors can be excised and the vessels repaired primarily. PMID- 16719199 TI - Deconstructing surgical education--teacher quality really matters: implications for attracting medical students to surgical careers. AB - Delineating those factors that enhance a student's surgical experience during medical school may be important to attracting "the best and the brightest" to surgery programs. Therefore, understanding the differences between an excellent and poor student evaluation from the student perspective is critical to surgical education, yet it remains ill defined. We concurrently assessed comprehensive student evaluations from a surgical clerkship over a 2-year period. The purpose of this study is to report the results of this audit. Two years of student directed evaluations were analyzed. Nine different surgical services were evaluated. Twenty-six data points were collected, including demographic, career, objective, and subjective information. Statistical analysis was performed using descriptive, chi2, and logistic regression tests. One hundred twenty-eight students rotated over 2 years, with 113 (88%) completing the assessment (61% men, 39% women). Men were more interested in surgical careers than women (4:1, P < 0.05). Medicine (22%) and surgical subspecialties (23%) were the most common career interests. Regression analysis demonstrated that age and gender were not predictors of outcome. The "highest rated and lowest rated" service were compared. Analysis demonstrated significant differences in three areas (operating room experience, and resident and faculty teaching.) The characteristics that separated the good teachers from the poor ones were the ability to challenge the student to think, providing useful feedback for their work, the ability to communicate ideas, and a positive attitude toward students/teaching. Student surgical evaluations underscore the role faculty and/or residents play in their education. PMID- 16719200 TI - Pancreaticoduodenectomy after neoadjuvant therapy in a Jehovah's witness with locally advanced pancreatic cancer: case report and approach to avoid transfusion. AB - Although the morbidity and mortality rates associated with pancreaticoduodenectomy (PD) have been improving over the past several decades, perioperative transfusions are often needed. Here, we review the preoperative planning and overall management of a Jehovah's Witness patient with locally advanced pancreatic cancer who would not accept blood transfusion. Management of this case is reviewed, along with the relevant literature regarding major surgery in the Jehovah's Witness population. The use of neoadjuvant chemoradiation was used successfully in locally advanced disease, allowing surgical resection. In addition, we outline a cogent strategy using pre-, intra-, and postoperative techniques to minimize blood loss and maintain hemoglobin at acceptable levels thereby preventing the need for transfusion. These strategies, once in place, may be able to reduce transfusions in all patients having major resections for malignancy. PMID- 16719201 TI - Prognostic importance of sclerosing variant in papillary thyroid carcinoma. AB - The diffuse sclerosing variant (DSV) of papillary thyroid carcinoma is known for its high aggressiveness, high incidence of lymph node metastases, and high incidence of pulmonary metastases, and thus its consequently poorer prognosis. In this study, we undertook a retrospective analysis of papillary thyroid carcinomas to assess whether DSV can be considered a predictive factor for prognosis. We performed a retrospective evaluation of the Department's database of patients with papillary thyroid carcinoma who had undergone total thyroidectomy from January 1992 to December 2000. Group I consisted of 83 DSV patients and Group II was 168 pure papillary carcinoma (PC) patients. A significant prevalence of multinodular thyroid disorder on diagnosis was found for PC (P < 0.05), whereas with DSV, there was a significantly higher prevalence of post-thyroiditis nodular thyroid disorder than with PC (P < 0.001). The incidence of laterocervical lymph node pathology on diagnosis was significantly higher for DSV (P < 0.05). In 3.6 per cent of PC patients and 15.7 per cent of DSV patients, we observed recurrences in the regional lymph nodes (P < 0.001). We found 1.2 per cent distant metastases in PC patients and 7.2 per cent in DSV patients (P < 0.05). One PC patient (0.6%) and three DSV patients (3.6%) died of tumor-related causes (P < 0.05). Our study demonstrated that diffuse sclerosing carcinoma leads to a poorer prognosis to the extent that its classification as an autonomous clinical pathological entity is justified. In conclusion, we can state that DSV is a form of papillary thyroid tumor characterized by its higher aggressiveness, diffuse intrathyroid growth, and high incidence of lymph node and pulmonary metastasis. Ultimately, this means a poorer prognosis. In the presence of risk factors indicating a possible increase in biological aggressiveness, adequate postoperative treatment and close follow-up become essential. PMID- 16719202 TI - Microdissection-based allelotyping: a novel technique to determine the temporal sequence and biological aggressiveness of colorectal cancer. AB - Pathologic staging in colorectal adenocarcinoma (CA) is based on the concept that the timing of metastatic tumor spread is directly related to the depth of the primary tumor invasion. To evaluate the temporal sequence of CA metastasis, we performed microdissection mutational profiling at multiple microscopic sites of primary and metastatic CA specimens. Twenty-one cases of CA were selected from fixed-tissue archives. Primary tumors were microdissected at the deepest point of invasion. Comparative mutational profiling for different genomic loci [1p36(CCM = cutaneous malignant melanoma], 3p26(OGGI = 8 oxoguanine DNA glycosylase), 5q23 (APC, MCC = mutated in colorectal cancer), 9p21(p16/CDKN2A = cyclin-dependent kinase 2A), 10q23(PTEN = phosphatase and tensin homolog [mutated in multiple advanced cancers 11), 12p12(K-ras-2 point mutation), 17p13(TP53), 18q25(DCC= deleted in colorectal cancer) was carried out on each microdissected tissue target using microsatellite loss of heterozygosity determination or DNA sequencing. All primary and metastatic sites of CA manifested acquired mutational change in 18 to 91 per cent of the genomic markers. In 15/21 (71%) cases, metastatic sites lacked a specific allelic loss seen in the corresponding primary tumor, indicating that the metastasis occurred before maximal depth of primary invasion. This was further supported by discordant mutational profiles between primary and secondary tumors, requiring divergent clonal evolution. This is the first report describing the temporal sequence and significance of sequential mutational acquisition in clinical tissue specimens with potential implications for a new molecular pathology approach to classify human cancer. PMID- 16719203 TI - A planned complex suicide: need for a high index of suspicion. AB - Unusual ways of committing suicide are reported in the medico-legal literature, but few refer to the so-called "planned complex suicide" (PCS). PCS, also termed primary combined suicide, is defined as the combination of two or more methods of injury by the suicidal person to prevent failure of a single method of suicide alone. In contrast, in secondary or unplanned complex suicide, the victim uses the second method of suicide only after failure of the first method.2 Combinations of shooting, hanging, burning, and/or poisoning have been used as suicidal methods. Sometimes the double use of one method such as simultaneous gunshot wound from two firearms or ingestion of two different toxic substances has been reported. This gives the suicidal person a high degree of protection against failure of one of the methods, as both modes of injury are relatively certain. This case report demonstrates the importance of the treating physician maintaining a high index of suspicion for the possibility PCS with the aim of avoiding a misdiagnosis that may be fatal. PMID- 16719204 TI - Gastric metastasis 14 years after mastectomy for breast lobular carcinoma: case report and literature review. AB - In planning treatment of a gastric neoplasm in a patient previously treated for lobular breast carcinoma, it is important to differentiate a primary gastrointestinal tract tumor from a metastatic form. We report a case of a breast lobular carcinoma metastatic to the stomach. The patient underwent a subtotal gastrectomy for symptomatic disease. Although gastric symptoms appeared 14 years after the breast carcinoma, immunohistochemical analysis of the surgical specimen helped to establish that the gastric lesion, thought to be primary, was effectively a metastatic repetition of the breast neoplasm. To better define treatment in a gastric neoplasm patient previously treated for breast carcinoma, the preoperative diagnosis should rule out a metastatic disease. The patient described received an adjuvant chemotherapy according to breast cancer protocol after gastric resection for symptomatic disease. The patient is still alive and undergoing chemotherapy for peritoneal carcinosis. PMID- 16719205 TI - Re: J-tip wire entrapment in a vena cava filter during central line placement. PMID- 16719206 TI - Selling sickness. PMID- 16719208 TI - What could the nematodes teach us about radiation protection? Plenty. PMID- 16719207 TI - Percutaneous vertebroplasty in patients with intractable pain from osteoporotic or metastatic fractures: A prospective study using quality-of-life assessment. AB - PURPOSE: Percutaneous vertebroplasty (PVP) is a minimally invasive outpatient procedure whereby vertebral compression fractures are stabilized by the injection of bone cement, or polymethyl methacrylate (PMMA). Rapid partial or complete pain relief can usually be achieved through this procedure. We prospectively evaluate the efficacy of PVP in the relief of pain and improvement in quality of life of patients with intractable pain from osteoporotic and metastatic fractures. MATERIALS AND METHODS: Patients with intractable pain from vertebral metastases (many resistant to palliative radiation therapy) and patients with intractable painful osteoporotic fractures were treated with parapedicular or transpedicular injection of PMMA. Plane X-rays, computed tomography (CT) scan, and magnetic resonance imaging (MRI) scan were performed on all patients. With a reflex hammer, percussion pain was correlated to the imaging abnormalities. The preplanning CT scan was used to calculate the exact entry point and angle of the bone-biopsy needle. All patients were assessed before and after the procedure for quality of life and amount of pain. The following measures were used: 1) the Edmonton Symptom Assessment System (ESAS), for global pain, nausea, tiredness, depression, anxiety, drowsiness, appetite, sense of well-being, and shortness of breath; 2) a site-specific pain score (SSPS); 3) the amount of analgesic intake in morphine equivalents in the last 24 hours; and 4) the Townsend Functional Assessment Scale (TFAS), ascertaining mobility. A postprocedural CT scan was performed the same day on all patients. Follow-up assessment consisted of a phone call at Days 1, 2, and 4 and Weeks 1, 2, 4, 8, and 12. Patients with recurrent back pain or complications were followed in the bone metastases clinic. RESULTS: Thirty patients (19 women, 11 men) were evaluated. Their median age was 68 years (range 31 to 87 years). Thirty procedures (n = 30) at 45 vertebral levels were performed; 13 were for pathologic fractures, and 17 were for osteoporotic fractures. SSPS showed a decrease of 2 or more levels in 88.5% of patients at 12 week follow-up. The mean SSPS with movement was 8.7 preprocedure and 1.8 postprocedure (P < 0.0001). Also, there was significant improvement in all 9 ESAS domains (P < 0.0004). The ingestion of analgesics in morphine equivalents showed a trend toward reduction post-PVP (P < 0.0599). When the patients with pathologic fractures were separated out, the reduction in ingestion of analgesics was significant (P < 0.0008). The TFAS demonstrated significant improvement in patient mobility and function. Extravertebral extravasation of cement was noted in 55.6% of the levels. We used a general linear mixed-model repeated-measures analysis of variance to analyze the data. CONCLUSIONS: PVP in osteoporotic and metastatic fractures significantly improved many patients' global quality-of-life scores and function by markedly decreasing their back pain and reducing their intake of pain medications. The procedure is safe, with no serious complications noted in our study. PMID- 16719209 TI - The interventional radiologist as "clinician": what does it mean? CanMEDS for the interventional radiologist. PMID- 16719210 TI - Detection rates in pediatric diagnostic imaging: A picture archive and communication system compared with a web-based imaging system. AB - OBJECTIVE: This prospective study assesses whether there are differences in accuracy of interpretation of diagnostic images among users of a picture archive and communication system (PACS) diagnostic workstation, compared with a less costly Web-based imaging system on a personal computer (PC) with a high resolution monitor. METHODS: One hundred consecutive pediatric chest or abdomen and skeletal X-rays were selected from hospital inpatient and outpatient studies over a 5-month interval. They were classified as normal (n = 32), obviously abnormal (n = 33), or having subtle abnormal findings (n = 35) by 2 senior radiologists who reached a consensus for each individual case. Subsequently, 5 raters with varying degrees of experience independently viewed and interpreted the cases as normal or abnormal. Raters viewed each image 1 month apart on a PACS and on the Web-based PC imaging system. McNemar tests were used to compare accuracy of interpretation across both imaging systems. Confidence intervals (CIs) were calculated for differences in the proportion assessed incorrectly on the PACS, compared with the Web-based PC imaging system. RESULTS: There was no relation between accuracy of detection and the system used to evaluate X-ray images (P = 0.92). The total percentage of incorrect interpretations on the Web based PC imaging system was 23.2%, compared with 23.6% on the PACS (P = 0.92). For all raters combined, the overall difference in proportion assessed incorrectly on the PACS, compared with the PC system, was not significant at 0.4% (95%CI, -3.5% to 4.3%). CONCLUSION: The high-resolution Web-based imaging system via PC is an adequate alternative to a PACS clinical workstation. Accordingly, the provision of a more extensive network of workstations throughout the hospital setting could have potentially significant cost savings. PMID- 16719211 TI - Canadian radiology residents' knowledge of sedation and analgesia: a web-based survey. AB - OBJECTIVE: An increasing number of procedures in the radiology department require the use of conscious sedation, with the agents often administered by the radiologist. We sought to determine the level of understanding of the nature and use of such agents in Canadian radiology residents. METHODS: A Web-based questionnaire was distributed to residents from 8 Canadian radiology residency programs. The questions concerned the pharmacology of common medications for conscious sedation, their indications and appropriateness for use, and the experience and attitudes of residents toward formal training in conscious sedation. RESULTS: A total of 178 surveys were dispersed and yielded an adjusted response rate of 51%. Most residents stated that they had not received any formal training in conscious sedation (65%) and were in favour (68%) of having such training. Although the residents typically correctly prescribed appropriate dosages of lorazepam (54%), midazolam (51%), and fentanyl (58.7%), excessively high dosages of midazolam were ordered by 15.9% of the residents. Knowledge regarding the onset of action and duration of commonly used medications was poor. Residents gave the correct response with regard to duration of action for lorazepam (23.8%), midazolam (31.9%), diazepam (15.9%), and fentanyl (28.6%). The correct responses to onset of action were as follows: for fentanyl, 22.2%; for midazolam, 19.1%; for lorazepam, 6.35%; and for diazepam, 11.1%. Residents were uncertain regarding the maximum dosage of local anesthetics that a patient could receive, with 1.5% and 20.6% correct responses regarding bupivacaine and lidocaine, respectively. CONCLUSION: Despite the recent publication of conscious sedation guidelines for nonanesthesiologists, this survey suggests that Canadian radiology residents are not receiving adequate training in the use of medications required for conscious sedation. PMID- 16719212 TI - Adenocarcinoma involving the uterine cervix: magnetic resonance imaging findings in tumours of endometrial, compared with cervical, origin. AB - PURPOSE: To determine the distinctive magnetic resonance imaging (MRI) features of cervical and endometrial adenocarcinoma that present clinically as a cervical mass. MATERIALS AND METHODS: From 1999 to 2002, 56 patients with adenocarcinoma on the initial biopsy of a cervical mass underwent MRI at our institution. Of these, 42 had a visible mass on MRI. Pathology review of all available tissue was the reference standard. A site of origin was determined by the pathologist in 38 of the 42 patients, and these were the cases evaluated; of these patients, 32 cases had adenocarcinoma and 6 had adenosquamous cancers. RESULTS: Findings were significantly more prevalent in patients with adenocarcinomas of endometrial, compared with cervical, origin for endometrial thickening (11 [73%] and 3 [13%], respectively; P = 0.0003), endometrial mass (11 [73%] and 1 [4%], respectively; P < 0.0001), endometrial cavity expansion by a mass (9 [60%] and 2 [9%], respectively; P = 0.001), and invasion of myometrium from endometrium (9 [60%] and 0, respectively; P < 0.0001). CONCLUSION: Adenocarcinomas of the endometrium that involve the cervix have MRI features that help distinguish them from primary adenocarcinomas of the cervix. PMID- 16719213 TI - Answer to case of the month #105. Methanol overdose. PMID- 16719214 TI - Answer to case of the month #106. Pelvic Digit. PMID- 16719215 TI - "I love you from the bottom of my heart". PMID- 16719216 TI - On a remarkable dental cement formulated by American Indians fifteen hundred years ago. PMID- 16719217 TI - Risk. PMID- 16719219 TI - The small business buy-sell agreement. PMID- 16719218 TI - Requirements for obtaining permits to administer anesthesia, etc. PMID- 16719220 TI - Dental water quality--the scientific evidence for current CDC guidelines. AB - Newly revised and updated guidelines from the Centers for Disease Control and Prevention recommend adoption of procedures to improve the microbiological quality of water used in dental treatment. These recommendations represent the consensus of experts from a wide range of scientific and clinical disciplines and are based on the best currently available evidence. Dentists and the manufacturers of dental equipment must consider the infection control, occupational health and risk-management implications of biofilm colonization in dental equipment, and take appropriate measures to provide water of appropriate microbiological quality clinical procedures. PMID- 16719221 TI - My personal Gulf War. PMID- 16719222 TI - A profile of female residents of Pennsylvania. AB - Specific information on the health of the women in Pennsylvania emphasizes the need for increased attention to needs of the more than six million female residents of the state. PMID- 16719225 TI - [Jubilee of health care system in B&H: 60 years of first medical school]. PMID- 16719223 TI - Formocreosol. PMID- 16719226 TI - [Early changes in rats cervical lymph nodes after pinealectomy]. AB - OBJECTIVES: Pineal gland acts as a reley where an anabolic activity be replaced to catabolic and conversely, to adapt a biological rhythm according to the organisme requirement. It is established that pineal extracts stimulate lymphopoiesis in the lymph node and had a radioprotective effects in lymph tissues. Lymph nodes reaction was investigated four weeks after pinealectomy, using qualitative and stereological. METHODS: One group of animals ( Wistar rats of both sex) was shame-pinealectomized (chirurgic treatment without pineal extirpation/control) and the other was submitted to surgical ablation of pineal gland.(experimental group). The animals were kept under the same ecologic conditions and they had free access to water supplies. Animals of the same group sacrificed at the same day between 10/14 hour to equal the effects of hormonal rhythm. The paraffin section died with HE (haematoxillin-eosin), PAS/alcian blue and impregnated by Bielschowsky method. For stereological analysis multipurpose M/42 test system was applied. Volume density of nodal structure and relative cellular distribution was evaluated. RESULTS: Cortical tissue of lymph nodes of pinealectomized rats was reduced, particularly the paracotical regions. Only in the female rats, decreased number of lymphocytes was observed in the cortex and modularly cords. Stereological analysis confirmed that increased volume density of the medulla is the result of medullar sinus dilatation. Under regressive changes of the cortical structures can be considered the involutary processes in germinal centers and reduction of the younger lymphocyte forms. Macrophage was activated and increased PAS + an inclusion was observed on the cortico/medullary junction and in the modularly cords. Reticular stroma was intensively impregnated and fibrils more coarsed and fragmented. DISCUSSION: Established changes in the lymph nodes of pinealectomized a rat was analogous with that observed after treatment with the pineal extracts but in the reverse means. It was evident that the structural dissociation of the reactivity had been established. Really, at one side we have involutive changes in lymphoid population and plasma cells but progressive in phagocytes and reticular cells. It is probably that increased concentration of corticosteroides and gonadal hormones after pinealectomy is responsible for these changes. CONCLUSION: Pineal gland-lymph nodes correlations established possibly via hypophyso-adreno-genital axis. PMID- 16719227 TI - [Cohort evaluation of treatment of smear positive tuberculosis patients]. AB - Direct observed treatment short course (DOTS) was introduced by National Tuberculosis (TB) Programme in our country in 1994. OBJECTIVE: to evaluate treatment outcome of tuberculosis patients by patient cohort group defining. PATIENTS AND METHODS: This study comprised 67 smear positive TB patients in 1998 and 86 patients in 1999. All patients were admitted in the University Clinic Sarajevo. RESULTS: Cure rate was achieved in 85.1% patients in 1998 and 91.8% in 1999.3% patients completed treatment in 1998 and 1.2% patients in 1999. Treatment success rate (cure rate and rate of completed treatment) was achieved in 88.1% patients in 1998 and 93% patients in 1999. Failure rate was 1.5 in 1998 and no failure was registered as treatment outcome in 1999. Treatment interrupted rate was registered in 5.9 patients in 1998 and 2.3 in 1999. There were no patients characterized as transferred out in 1998, but the rate of these patients in 1999 was 1.2 . Mortality rate was 4.5 in 1998 and 3.5 in 1999. CONCLUSION: DOTS expansion has contributed the increased percentage of successfully cured patients. PMID- 16719228 TI - [Analysis of surgical approach to diagnosis and therapy in solitary pulmonary nodules]. AB - OBJECTIVE: Descriptive-analytic appraisal of validity of surgical approach to diagnosis and therapy in solitary pulmonary nodules (SPNSs). TASK: to establish justification of surgical removing of SPNs which etiology is unclear after complete previous clinical investigation. PATIENTS AND METHODS: In the period between 1 January 1999 and 1 January 2004, included in this investigation were all patients with SPN of unclear origin who were referred to the Clinic for Thoracic Surgery-UCC Sarajevo after complete previous clinical investigation. RESULTS: We operated 60 patients with proved SPN. The malignancy was found in 44/60 (73,3%) patients. There were established following types of malignancy carcinoma planocellulare in 54,5%, adeno carcinoma in 36,5% carcinoma bronchioloalveolare in 4,5% and tumor carcinoides in 4, % patients. We did following types of operation atypical (wedge) resection in 20 (33,3%), lobectomy in 36 (60,0%) and pneumonectomy in 4 (6,7%) patients. There were neither intra- operative complications nor intra operative mortality. Postoperative morbidity include: pleural emphysema in 1 (1,65%) and stress ulcers in 1 (1,65%) patients. There was not early postoperative mortality (until 30 days after operation). CONCLUSION: The malignancy was the most frequent cause of the SPNs. Operative and postoperative morbidity and mortality are relatively negligible. High percentage of the malignancy in SPNs, negligible operative and post operative morbidity and mortality justify aggressive surgical approach to diagnosis and therapy in SPNs and point out to the necessity for early removing of this kind of pulmonary lesion. PMID- 16719229 TI - [Critical congenital heart diseases in Tuzla Canton area]. AB - Critical congenital heart diseases (CHD) represent extensive and complex possibilities changes of morphology and homodynamic on the heart, which vital jeopardize child at the time of birth so as at the first weeks of its life or at the first months of its life too. The purpose of this study is to determine frequency, structure and the mode of medical treatment critical UAS in the Tuzla Canton area to harmonies with experiences and results we have reached in order to treat children who have congenital heart disease. There was discovered CHD in the 493 (8,15 per 1000 live-born) from 60.493 live-born children in Tuzla Canton area during the 10-year period start at 01.01.1995. Critical congenital heart disease was discovered in 142 children what represent total prevalence of 2,35 per 1000 live-born children. Yearly prevalence was pretty consistent in observed period of the time and it was from 1,36 to 3,24 per 1000 live born children. The most frequent UAS was complete atrio ventricular canal, which has proved in the 22,4% critical UAS. The more then one-third (36,6 %) children with critical UAS have had extra cardiac anomalies and syndromes. The balloon septostomy was performed in only two of our patients, in 41 (28,8 %) cardiac surgery was performed. Only one of the children underwent surgery in the neonatal period. In 8 patients who were no surgery treated developed pulmonary hypertension, one of them had bacterial endocarditic. Of the total 142 children with critical CHD 99 (69,7%) died. The 92 of them died before surgery, 7 of them died after the surgery; 3 of the 7 in the early after surgery period; 4 of the 7 died because of complications in a later period. Timely discovering critical UAS in Tuzla Canton area is failure in the area prenatal diagnosis. The treatment of patients is inappropriate and without possibilities of emergency invasive cardiac treatment and surgical treatment. PMID- 16719230 TI - [Diagnostical values of antibody on A60 antigen in diagnostic of tuberculosis]. AB - Aim of this research was to investigate diagnostic value of discovering of antibody on A60 antigen in patients who were tested for presents of Mycobacterium tuberculosis in there biological samples. We tested a samples of sputum, gastric juice, urine, cerebrospinal fluid and punctate from group of 353 patients who were suspected for tuberculosis. In all patients we were looking for antibodies classes A60 antigen. We used immune chromatographic "Hexagon TB" test, Germane company "Human Geselschaft fur Biochemica und Diagnostica". From 353 patients we found 58 (16.43%) patients with positive BK, 79/22,38%) patients with positive Lowenstein culture and 122 (34,55%) patients with antibody in sera on A60 antigen. Patients who were BK and Lowenstein positive, have had antibody in 94,23% cases, Patients who were BK negative and Lowenstein positive have had antibodies in 70,37% cases and patients who were BK negative and Lowenstein negative have had antibody in 19,03% cases. Patients with BK positive and Lowenstein negative results have had antibody in 50,00% cases. Difference between results is significant (p<0,01). From 122 patients with positive antibodies, 52 were BK positive and 68 have had positive Lowenstein cultures. From 231 patients with no antibody, just 6 were BK positive and 11 Lowenstein positive. In 62 patients with positive antibodies, were BK and Lowenstein negative. We confirmed that antibody on A60 antigen in microbiological positive patients are more often then in microbiological negative patients (p<0,001). PMID- 16719231 TI - [Breast Paget disease: morphologic substrate and Paget cells characteristics]. AB - Authors analyzed 143 cases of breast Paget disease with special emphasis on basic tumor process and on immune histochemical characteristics of tumor cells. Breast Paget disease represent special clinical presentation of the breast tumor. The most often tumor in the breast are ductal carcinoma in situ, ductal carcinoma and in rare instance lobular carcinoma and lobular carcinoma in situ. There are several clinical combinations with or without involvement of axillary's lymph nodes. The best prognosis is in cases with in situ as underlying process. All other combinations have worse prognosis in relation to the combination with in situ lesion. The middle ages of patients with Paget disease is 10 years higher than the ages without areola involvement. Paget cells show the same immune histochemical characteristic as tumor cells. The authors propose mastectomy instead central segmentectomy. PMID- 16719232 TI - [Evaluation of the tumor of the pituitary gland (hypophysis) between 2000-2005. On the intensive care of The Endocrinology Clinic of the Clinical Center of the University of Sarajevo]. AB - INTRODUCTION: Tumors of the pituitary gland can be adenomas, and more often, cancers, representing 10-15% of intracranial neoplasm. We make a difference between them according to their size, expansion, hormone's activity and local and endocrinological manifestations. The goal of the research was to reveal the presence of tumors of the pituitary gland in the period 2000-2005 among patients hospitalized on the Intensive Care of the Endocrinology Clinic. PATIENTS AND METHODS: Data from anamnesis of all patients with verified tumor of the pituitary gland between 2000.-2005 were used in the research work. RESULTS: Total number of patients with the tumor of pituitary gland was 40, 29 women and 11 men, and 35 % of patients were aged between 45 and 60. Main symptoms: headache -16, eye trouble -17, sexual dysfunction -8, growth of acres -5, and others symptoms -8 patients. There were 23 (57,5%) cases of micro adenoma and 17 (42,5 %) macro adenomas. From the total number of patients 23 were operated, 8 micro adenomas and 15 macro adenomas. The number of secreting adenomas was 25 (62,5 %) and 15 (37,5 %) were non-secreting. Secreting adenomas were: prolactinomas--14, acromegalies -7, Cushing syndromes -2, TSH producing -2. Non-secreting adenomas were: craniopharyngeomas--11, meningiomas -2, and 2 non-secreting micro adenomas. Most of prolactinomas were treated with drugs, and 3 cases of prolactinoma were operated. 6 cases of acromegaly were operated and 1 micro adenoma is still under medical supervision, with the therapy of bromocriptine. 3 of those patients have the residuum, and 4 of them have high level of hGH. 11 craniopharyngeomas were operated and all patients, except one, are under substitution therapy. 2 meningiomas were operated and those patients are also under substitution therapy. 1 case of TSH producing tumor was operated, but residuum and hyperthyroidism are still presents, and the micro adenoma, for the hyperplasia reason, disappeared with adequate substitution therapy. The Cushing syndrome, confirmed with MRI diagnostics, wasn't operated; it remained under conservative therapy, and in the other case, where we didn't isolate a pathological substrate, the patient exited. CONCLUSION: Tumors of the pituitary gland we mainly treat applying surgery methods, together with post operational substitution therapy or additional therapy with drugs. Only prolactinomas are tumors which are successfully treated with drugs. PMID- 16719233 TI - Impact of rosiglitazone on glycaemic control, insulin levels and blood pressure values in patients with type 2 diabetes. AB - INTRODUCTION: The main metabolic defect responsible for hyperglycaemia in type 2 diabetes is insulin resistance and impaired B-cell function. Currently much more medications which combat impaired B-cell function are available then those which might inhance insulin sensitivity. A novel oral hypoglycaemic agent, rosiglitazone, targeting insulin resistance, not yet registered in our country, has been available in majority of Europian countries. The main aim of the study was to evaluate the efficacy of rosiglitazone in the treatment of patients with type 2 diabetes patients compared with glibenclamide monotherapy. METHODS: A total sample of 20 diabetic patients, who had not achieved satisfactory control with dietary modifications, was selected. 10 patients were randomised to rosiglitazone treatment in daily dosage of 4 to 8 mg (RG group) while the rest of the patients were receiving glibenclamide in the dosage of 3.5-10.5 mg daily (GC group). All the patients had their fasting and postprandial glycaemia, HbAlc, fasting insulin and blood pressure measured at baseline and 3 months after initiation of the treatment. Possible increase of the liver enzymes in the RG group was followed up. RESULTS: There was a significant reduction in glycaemic parameters in both groups in comparison to baseline values. However fasting insulin levels in the RG group were significantly reduced while slight but not significant increase was noted in GC group. Blood pressure values were significantly reduced in the RG group while no changes were noted in the GC group. CONCLUSION: Results indicate significant improvement of glucose utilisation as a consequence of reduced insulin resistance in the RG group. Treatment with rosiglitazone lead to improved insulin sensitivity which may predict longer term preservation of B-cell function. In addition, this novel agent also have favourable impact to blood pressure values which is of exceptional importance in the complex management of type 2 diabetic patient. PMID- 16719234 TI - [Bladder, bowel and sexual dysfunction in patient with mulitple sclerosis]. AB - Bladder, bowel and sexual dysfunction are not rare in multiple sclerosis (MS). The most frequent bladder disorders are in urgency, frequency as well as retention. In this study we analyzed bladder dysfunction (urgency, frequency and retention), defecation (constipation and incontinence) as well as sexual problems (libido, erection and vaginal lubrication) in patients with relapsing-remitting form of multiple sclerosis (MS) depending on disability status. Concerning Expanded Diasability Status Score (EDSS), patients have been divided in two groups: EDSS 0-4.5. (Group A), and EDSS 5.0-10.0 (group B). We analyzed 60 patients: 35 (58.3%) women, and 25 (41.7%) men. In both group A and B, with no bladder disorder was 18 patients (30%). In group A: 15 (25%); in group B: 3 (5%). With urgent incontinence were 36 (60%) patients - group A: 19 (31%); group B: 17 (28%). With retention were 6 (10%) patients - group A: 4 (6.6%); group B: 2 (3.3%). There is no statistical significant difference in number of patients with and with no urinary problems in both groups (p>0.05). Of all analyzed patients in group A and B with no urinary disorders were 9 men (15%) and 9 women (15%). With urinary problems were 16 men (26.7%) and 26 women (43.3%). Statistically it is significant higher number of women than men (p<0.05). With no defecation problems were 54 (90%) patients - in group A: 36 (60%); in group B: 18 (30%). With constipation were 5 (8.3%) - in group A: 2 (3.3%); in group B 3 (5%). In group B one patient (1.6%) had incontinence. There is no statistical significant difference in number of patients with and with no bowel elimination dysfunction in both groups p>0.05). 12 (20%) men had no bowel elimination problems and 24 women (40%), but 10 (16.6%) men and 8 (13.3%) women had the bowel elimination dysfunction. There is no statistical significance (p>0.05). No libido disturbance had 44 (73.3%) patients. In the group A: 35 (58.3%); in group B: 9 (15%). 16 (26.7%) patients had the libido disturbances. In group A: 3 (5%); in group B 13 (21.6%). Statistically there is significant lower number of patients with libido problems in group A (p<0.05). No vaginal lubrication had 21 (35%) women. In group A: 18 (30%); in group B 3 (5%). Vaginal lubrication disturbance had 14 (23.3%). In group A: 7 (11.6%); in group B 7 (11.6%). Statistically it is significant lower number of women with no lubrication in group B (p<0.05). No erection problems had 11 (18.3%) men. In group A: 9 (15%); in group B 2 (3.3%). Erection disturbance had 14 (23.3%). In group A: 4 (6.6%); in group B: 10 (16.6%). There is statistical significant lower number of men with erection problems in group A. (p<0.05). Of all analyzed patients in both groups A and B 18 men (30%) and 26 (43%) women had no problems with libido and 7 (11.6%) men and 9 (15%) women had the problems. There is no statistical significance in sex distribution (p>0.05). 11 (18.3%) men had no erection problems, 14 (23.3%) had the problems. 21 (35%) women had no vaginal lubrication problems, 14 (23.3%) women had the problems. Erection disturbances are not statistically significant to vaginal lubrication problems (p>0.05). PMID- 16719235 TI - [Clinical audit as method of quality improvement of healthcare in patients with diabetes, stroke and in cesarean section]. AB - INTRODUCTION: Clinical audit is used as a term for any kind of audit leaded by professionals in health care, and should not be complicated or unpleasant job. AIM: The aim of this work is to demonstrate the variations in practice between doctors working in a health care organization and the institutions (health centers - DZ and hospitals). Variations in practical work show the difference in quality of work between the doctors and the institutions, as well as variations in the use of resources and expenditure costs. METHODS: Three different questionnaires had been developed for three different clinical entities: diabetes, cesarean section, and stroke. The questionnaires had been created in collaboration with experts from each of those medical entities and based on research of literature, and valorized by the questionnaire for creating audits and calculations of ACQ (Audit Criteria Questionnaire) score. RESULTS: Diabetes- The hugest disproportion is in not measuring (none evidencing) regular annual screenings for retina and foot, but albumin and creatinin are being controlled in very high percentage. Stroke: In hospital A in checked medical records side of cerebral lesion was evidenced regularly, what was different in hospital B, while in hospital B 100% of swallowing disorders were evidenced. Cesarean section: In hospital A the average period of preoperative length of stay is four days, while in hospital B it is 0.4 days, while the postoperative length of stay for both hospitals is 6 days. DISCUSSION: Clinical audit is a systematic analysis of quality of health care and as per the Accreditation standards for hospitals, published by the Federal Agency for Health Care Quality and Accreditation-AKAZ; it is mandatory to perform the clinical audit in 3-5 areas twice, in the year precedes the accreditation survey. CONCLUSION: Using small sample, authors proved big variations in practice among doctors and among health institutions as well. It has direct consequence on quality and costs and can be solved using AKAZ accreditation standards. It is very difficult to improve the quality of health care without regularly performed and documented clinical audits. PMID- 16719236 TI - [Controversies about human cloning]. AB - Human cloning is one of the greatest scientific and research challenges that human kind faced so far. As much as we have reasons for excitement in front of achievements of the genetic engineering, also are great the fears about it. There is inevitable question: is the man aware where can cloning as an attack on a "natural laws for the life reproduction" can lead, or to say its impact on evolution diversities within kind that we belongs to. In this paper we have focus on finding the answer on the last question, with application of a multidisciplinary approach, which means not only scientific, but also ethical, as well as theology. PMID- 16719237 TI - [Craniofacial surgical treatment of the gigantic plasmacytoma of the skull base: case report]. AB - The complex craniofacial surgery of solitary bone plasmacytoma (SBP) of the skull base with excellent postoperative outcome was presented. This surgical management was result of the team approach and strategy of neurosurgeon (KD) and maxillofacial surgeon (TM). SBP of the skull base is very rare condition which should be treated by radical microsurgical resection and postoperative radiotherapy. PMID- 16719238 TI - [Vaccine for prevention of type 1 diabetes mellitus]. AB - Diabetes mellitus type 1, known as insulin dependent diabetes mellitus (IDDM), results from a chronic autoimmune destruction of the insulin secreting pancreatic beta cells. This autoimmune process is marked by circulating auto antibodies to beta cell antigens. GAD 65 (Glutamic Acid Decarboxylase) auto antibodies are present in 70-80% of newly diagnosed patients with type 1 diabetes, and it can be detected many years before clinical onset of the disease. New knowledge in the imunopathogenesis have given expectation that imunotherapy will become routine manner for cure of diabetes mellitus type 1. PMID- 16719239 TI - [Stents in treatment of laryngotracheal stenoses]. AB - Endo larynges tracheal stenoses due to endo luminal or extra luminal processes of benign or malignant character in majority of cases demand the resection of a stenosed part. The use of a stent represents a necessity. In this paper we analyzed 10- year experience of the ENT Clinic of the University Clinical Center Sarajevo regarding the use of stents. 26 patients underwent surgery in a period 1995-2005. Prolonged endo tracheal intubations are encountered as a primary cause of stenoses (73 %). Infection and granulations are noted as a complication in 15,3% of cases. Stent is removed in 16 patients within 3 months and in 7 patients after 12 months. PMID- 16719240 TI - [Aortic stenosis and atrial septal defect in pulmonary hypertension caused by aorto-pulmonary fenestration]. AB - This article writes about a 37-year-old patient with Eissemanger's syndrome. The catheterization was done when he when he was 7 years old and was diagnosed as follows: VSD, ASD and pulmonary hypertension. After repeated catheterization when patient was 23 years old the final diagnosis was established: Fenestra aortico pulmonalis, VSD, ASD, aortic stenosis gr II and pulmonary hypertension. The patient has been treating conservatively PMID- 16719241 TI - [IABP and off pump revascularization as prefered method in curement of patients with "low cardiatic output" syndrome]. AB - Treatment of patients with "low output" syndrome is very complex as well as surgery method as well as post operative treatment. Surgery requests experience and well trained surgical team which is able to produce off pump revascularization of myocardium and in short period of time eliminate global contra effects of ischemia already damaged myocardium. In post surgery period treatment of these patients demands maximal medication support and use of IABP which implantation, in this case, been very useful. PMID- 16719242 TI - Buprenorphine: effective treatment of opioid addiction starts in the office. PMID- 16719243 TI - Diagnosing allergic rhinitis: skin test vs. IgE testing. PMID- 16719244 TI - The diminishing role of FPs in caring for children. PMID- 16719245 TI - Carbamazepine for acute and chronic pain. PMID- 16719246 TI - Screening for depression. PMID- 16719247 TI - Dietary advice to lower cardiovascular risk. PMID- 16719248 TI - Hypertension in children and adolescents. AB - The development of a national database on normative blood pressure levels throughout childhood has contributed to the recognition of elevated blood pressure in children and adolescents. The epidemic of childhood obesity, the risk of developing left ventricular hypertrophy, and evidence of the early development of atherosclerosis in children would make the detection of and intervention in childhood hypertension important to reduce long-term health risks; however, supporting data are lacking. Secondary hypertension is more common in preadolescent children, with most cases caused by renal disease. Primary or essential hypertension is more common in adolescents and has multiple risk factors, including obesity and a family history of hypertension. Evaluation involves a thorough history and physical examination, laboratory tests, and specialized studies. Management is multifaceted. Nonpharmacologic treatments include weight reduction, exercise, and dietary modifications. Recommendations for pharmacologic treatment are based on symptomatic hypertension, evidence of end-organ damage, stage 2 hypertension, stage 1 hypertension unresponsive to lifestyle modifications, and hypertension with diabetes mellitus. PMID- 16719249 TI - Managing opioid addiction with buprenorphine. AB - Legislation has enabled physicians to treat opioid-dependent patients with an office-based maintenance program using buprenorphine, a partial mu-opioid receptor agonist. Clinical studies indicate buprenorphine effectively manages opioid addiction. Buprenorphine is more effective than placebo for managing opioid addiction but may not be superior to methadone if high doses are needed. It is comparable to lower doses of methadone, however. Treatment phases include induction, stabilization, and maintenance. Buprenorphine therapy should be initiated at the onset of withdrawal symptoms and adjusted to address withdrawal symptoms and cravings. Advantages of buprenorphine include low abuse potential and high availability for office use. Disadvantages include high cost and possible lack of effectiveness in patients who require high methadone doses. Most family physicians are required to complete eight hours of training before they can prescribe buprenorphine for opioid addiction. PMID- 16719250 TI - Information from your family doctor. Opioid addiction: what you should know. PMID- 16719251 TI - Diagnosing rhinitis: allergic vs. nonallergic. AB - Allergic rhinitis, the most common type of rhinitis, generally can be differentiated from the numerous types of nonallergic rhinitis through a thorough history and physical examination. Allergic rhinitis may be seasonal, perennial, or occupational. The most common cause of nonallergic rhinitis is acute viral infection. Other types of nonallergic rhinitis include vasomotor, hormonal, drug induced, structural, and occupational (irritant) rhinitis, as well as rhinitis medicamentosa and nonallergic rhinitis with eosinophilia syndrome. Since 1998, three large expert panels have made recommendations for the diagnosis of allergic and nonallergic rhinitis. Allergy testing (e.g., percutaneous skin testing, radioallergosorbent testing) is not necessary in all patients but may be useful in ambiguous or complicated cases. PMID- 16719252 TI - Testosterone treatments: why, when, and how? AB - Testosterone treatment is controversial for men and even more so for women. Although long-term outcome data are not available, prescriptions for testosterone are becoming more common. Testosterone is used primarily to treat symptoms of sexual dysfunction in men and women and hot flashes in women. Potential benefits include improved libido, increased bone mass, and increased sense of well-being. In individuals with human immunodeficiency virus infection or other chronic diseases, testosterone has been shown to improve mood and energy levels, even in patients with normal testosterone levels. Testosterone can be administered by injection, patch, topical gel, pill, or implant. Side effects in men include polycythemia and acne. Side effects in women include acne, hepatotoxicity, and virilization and usually only occur when testosterone is used in supraphysiologic doses. Long-term studies of the effects of testosterone on prostate cancer, breast cancer, and heart disease have not been completed. Mammograms and monitoring of prostate-specific antigen, hematocrit, and lipid levels are recommended for patients taking testosterone. PMID- 16719253 TI - Information from your family doctor. Testosterone therapy: what you should know. PMID- 16719254 TI - Screening for speech and language delay in preschool children: recommendation statement. PMID- 16719255 TI - Nocturnal enuresis. PMID- 16719256 TI - Swollen masses in the nose. PMID- 16719257 TI - Determining eligibility for gastric bypass surgery. PMID- 16719258 TI - [The uses of clinical studies]. PMID- 16719259 TI - [Number needed to treat]. PMID- 16719260 TI - [Ixodes ricinus. Health risks and approaches in prevention]. AB - Ticks belong to the parasitical mites. There are more than 1000 different species worldwide which do need blood of warm-blooded organisms for their own development. As ticks are well known to be vectors of several pathogens which could cause insidious infectious diseases, they are important in human and veterinarian medicine as well. It is not possible to control ticks in nature. Therefore the first step should be an effective prevention. In case of a bite the tick must be removed as soon as possible. Although Ixodes ricinus is more or less responsible for all health risks caused by ticks in Germany, further species like Rhipicephalus sanguineus, Dermacentor marginatus and Argas reflexus must not be forgotten, as these ticks have appeared more frequently in the recent past and they can be vectors of several diseases as well. This paper describes first and foremost the biological behaviour and development of Ixodes ricinus and the medical importance of this tick in human and veterinarian medicine. PMID- 16719262 TI - [Danger from stick-on tattoos]. PMID- 16719261 TI - [Interaction between an antiinfective agent and an immunosuppressant after liver transplantation]. AB - Interactions between immunosuppressants and antiinfectives are most important in pharmacotherapy. Patients receiving this drug combination must be monitored carefully. Individual dose adaptation and therapeutic drug monitoring (TDM) according to the SOAP scheme (subjective data, objective data, analysis, plan) are exemplified by the case report of a 47-year-old patient who had undergone a liver transplantation. PMID- 16719263 TI - [Anticoagulants. Self management is safer]. PMID- 16719264 TI - [Smoking cessation. Reduced mortality after 14 years]. PMID- 16719265 TI - [ Does lowered folic acid from alcohol increase breast cancer risk?]. PMID- 16719266 TI - [Oral cancer. Cyclooxygenase inhibitors for prevention: beneficial or risky]. PMID- 16719267 TI - [Adolescence. Body weight is influenced by physical activity]. PMID- 16719268 TI - [Treatment of meteorism]. PMID- 16719270 TI - [Homeopathy is placebo]. PMID- 16719269 TI - [Treatment with the first glycylcycline--tigecycline]. PMID- 16719271 TI - [Benefits and risks of drug industry "sponsorship"]. PMID- 16719272 TI - Roberts syndrome: study of 4 new Rgyptian cases with comparison of clinical and cytogenetic findings. AB - Roberts syndrome is a rare autosomal recessive genetic disorder (MIM 268300). It is characterized by pre and postnatal growth retardation, severe shortening of limbs with radial defects, oligodactyly and characteristic facial features. The present study reports 4 new cases of Roberts syndrome from 3 families presenting variable phenotypes. Patients were thoroughly investigated clinically and cytogenetically. By reviewing literature, we compared our cases to those previously reported. The rating severity system proposed by Van den Berg and Francke (30) was applied to correlate the phenotypic and cytogenetics changes. We observed more severe reduction defects in the upper limbs than in the lower limbs. While the main reduction defects in the upper limbs involved the thumb and radius ranging to phocomelia, absent or severely hypoplastic fibula was the main lower limb involvement. We emphasize this finding in the present investigation. Heterochromatin repulsion of chromosomes derived from Roberts syndrome patients is a characteristic cytogenetic abnormality. It was a constant finding in our studied patients demonstrated by DABI stain which supports the possibility that mutations in Roberts syndrome lie in centromere related proteins which may also play a role in body patterning. This was proved recently by Vega et al. (31). Application of the clinical rating score and its correlation with cytogenetic changes showed negative results. Cytogenetic studies in normal obligatory heterozygotes parents showed no changes. Phenotypic variability within the same family as well as between different families was observed. The ascertainment of 4 cases with Roberts syndrome from 3 Egyptian consanguineous families during one year in our department may indicate a high frequency of the Roberts syndrome allele among Egyptians. This confirms the need for molecular studies for early and accurate prenatal diagnosis to prevent such dramatic malformation syndrome. PMID- 16719273 TI - Six cases of cryptic subtelomeric translocations in four families: the use of subtelomeric FISH probes as a diagnostic tool. AB - Finding the diagnosis in children with mental retardation and a normal karyotype, whether or not associated with dysmorphic features, is important for defining an eventual syndrome and for genetic counselling of the families. Telomeric re arrangements may be a common and underestimated-to-date cause of non-syndromic mental retardation. Using a FISH-based approach combining subtelomeric probes, we report the detection of 4 cases of cryptic translocations t(2;10)(p25.3;q26.3), t(4;17)(p16.2;q25), t(4;20)(p16.2;q13) and t(5;7)(p15.3;q36) associated with MR and dysmorphic features. We discuss the usefulness of subtelomeric FISH in children with unexplained delayed psychomotor development, when the genetic cause remains unknown and the karyotype is normal. PMID- 16719274 TI - Congenital anomalies and developmental delay in a boy with double chromosome 6 derived supernumerary marker. AB - The frequency of small supernumerary marker chromosomes has been estimated to approximately 0.45 per 1000 newborns. They are usually seen as single marker chromosomes in a mosaic state. Two cytogenetically identical markers have been observed only occasionally. We report on a boy, with congenital heart defect, neonatal hypotonia, hypogenitalism, delayed psychomotor development and mild dysmorphic facial features. The GTG karyotype performed on peripheral blood lymphocytes revealed a mosaic male karyotype with three cell lines. One cell line had a normal karyotype. In the other two either single or double chromosome 6 derived supernumerary markers were present, leading to partial trisomy or partial tetrasomy of chromosome 6, respectively. PMID- 16719275 TI - Wolf Hirshhorn syndrome in a case of ring chromosome 4: phenotype and molecular cytogenetic findings. AB - Ring chromosome 4 associates concomitant loss of the telomeric 4p and 4q regions and leads to variable clinical manifestations depending on the size of the deleted chromosomal material. We report on a patient with ring chromosome 4, showing the Wolf-Hirshhorn Syndrome (WHS) phenotype and minor symptoms of distal 4q deletion syndrome; the severity of the signs of WHS masks the symptomatology of the 4q deletion syndrome. The absence of seizures despite the absence of the specific 4p16.3 region with haploinsufficiency of the LETM1 gene is striking. The double telomeric deletion due to the ring chromosome formation confirmed by FISH has been rarely described in WHS. PMID- 16719276 TI - Waardenburg syndrome in the Turkish deaf population. AB - Waardenburg Syndrome (WS) is an autosomal, dominantly inherited disorder that accounts for more than 2% cases of congenital deafness. The aim of this study is to determine the WS incidence among deaf pupils. Dysmorphological examination was performed on 720 children who were attending 7 special schools in Turkey and who had hearing disabilities. All subjects in the study were examined for WS diagnostic criteria. We detected 49 patients (6.8%) with WS among the 720 children examined. Six patients had WS type 1 (12.2%) and 43 had type 2 (87.8%). We observed 2 to 5 major diagnostic criteria for WS. Out of all the subjects in the study, only two patients have deaf first degree relatives. All subjects had been previously examined by physicians for deafness but none of them had been then diagnosed to have Waardenburg Syndrome. Instead, they were all misdiagnosed as to have nonsyndromic deafness. Awareness of WS diagnostic criteria by the physicans will provide accurate diagnosis for many deaf pupils and their first degree relatives who are able-to-hear WS patients and whose children are at risk for deafness. PMID- 16719277 TI - A rare case of aniridia and balanced translocation (5;11) (p15.3;q22) arising in the same subject: a challenge for genetic counseling. AB - The authors report on a case of isolated aniridia caused by haploinsufficiency of the PAX6 gene, which is located on 11p13, and a balanced translocation t(5;l1)(p15.3;q22) inherited respectively from his father and his mother. Due to the coincidence of two abnormalities in the same chromosome, the segregation of the mutant allele leading to aniridia and of the chromosomes involved in the translocation are not independent events. Considering that both monosomy and trisomy for 11q22-qter are unviable, his offspring may inherit either the PAX6 mutation or the balanced translocation. However, depending on the occurrence of crossing over, there is a possibility for him to have normal offspring; on the other hand, he may also father children with both anomalies. This unusual case, in which the proband has a presumably very low chance of completely normal offspring, turned to be a challenge for genetic counseling. PMID- 16719278 TI - Spectral karyotyping and fluorescence in situ hybridization analysis of de novo partial trisomy 7p (7p21.2-->pter) and partial monosomy 12q (12q24.33-->qter). AB - An 8-year-old boy presenting with hypotonia, moderate mental retardation, developmental delay, and psychomotor retardation is reported. Magnetic resonance imaging of the brain at age 3 years revealed a Dandy-Walker variant. Cytogenetic analysis of the peripheral blood revealed a derivative chromosome 12 with unknown additional material attached to the distal region of the long arm of chromosome 12. The parental karyotypes were normal. Spectral karyotyping (SKY) using the 24 color SKY probes and fluorescence in situ hybridization (FISH) using the specific 7p, 7q, 12p, and 12q telomeric probes confirmed a duplication of distal 7p and a deletion of terminal 12q. The karyotype of the proband was designated as 46,XY.ish der(12)t(7;12) (p21.2;q24. 33)(SKY+, 7pTEL+, 12qTEL-). The present case provides evidence for the association of partial trisomy 7p (7p21.2-->pter) and partial monosomy 12q (12q24.33-->qter) with a cerebellar malformation and the usefulness of SKY and FISH in the identification of a de novo aberrant chromosome resulting from an unbalanced translocation. PMID- 16719279 TI - Prenatal diagnosis of Meckel Gruber syndrome presenting with renal agenesis: report of a case. PMID- 16719280 TI - Lack of significant association between chromosome Y microdeletion and varicocele in Turkish patients. PMID- 16719281 TI - Ellis van Creveld syndrome associated with bilateral tibial exostoses. PMID- 16719282 TI - Unilateral symbrachydactyly of the foot. PMID- 16719283 TI - Molecular cytogenetic analysis of de novo partial monosomy 4p (4p16.2-->pter) and partial trisomy 8p (8p23.2-->pter). PMID- 16719284 TI - Recurrent Galloway Mowat syndrome associated with abnormal prenatal sonographic findings. PMID- 16719285 TI - Detection of euchromatic variants and unusual C band heterochromatin variants at genetic amniocentesis. PMID- 16719304 TI - [Structure and function of actin]. PMID- 16719305 TI - [Regulation of actin polymerization by Rho family]. PMID- 16719306 TI - [Role of cofilin phosphorylation in cell migration]. PMID- 16719307 TI - [Actin dynamics]. PMID- 16719308 TI - [Molecular process of the dynamic instability of microtubule]. PMID- 16719309 TI - [Structure and function of microtubule-associated proteins]. PMID- 16719310 TI - [Microtubule plus-end-binding proteins (+TIPs)]. PMID- 16719311 TI - [Intermediate filaments: the mechanisms for the assembly and the physiological functions]. PMID- 16719312 TI - [Evolution and mechanical models of the myosin superfamily]. PMID- 16719313 TI - [Regulation of myosin II in cell division and cell migration]. PMID- 16719314 TI - [Versatility of kinesins in microtubule interaction]. PMID- 16719315 TI - [Roles of kinesins in mitosis]. PMID- 16719316 TI - [Unique role of LIS1 and NDEL1 as a regulator of cytoplasmic dynein]. PMID- 16719317 TI - [Nuclear structure: its molecular basis and dynamics]. PMID- 16719318 TI - [Molecular mechanisms of organogenesis regulated by ECM: the role in epithelial branching morphogenesis]. PMID- 16719319 TI - [Inside-out integrin activation links micro- to macro-dynamics]. PMID- 16719320 TI - [Integrin signaling and Src family kinases]. PMID- 16719321 TI - [Biophysical mechanisms of neurite growth mediated by the cell adhesion molecule L1]. PMID- 16719322 TI - [Structure and function of cadherins]. PMID- 16719324 TI - [The roles of beta-catenin/plakoglobin in cadherin-mediated cell-cell adhesion]. PMID- 16719323 TI - [Alpha catenin and cadherin]. PMID- 16719325 TI - [Roles of IQGAP1 in E-cadherin-mediated cell-cell adhesion]. PMID- 16719326 TI - [Molecular mechanisms of cadherin trafficking]. PMID- 16719328 TI - [Overview of plasma membrane/cytoskeleton interaction]. PMID- 16719327 TI - [Signal crosstalk between cell-substratum adhesion and cell-cell adhesion]. PMID- 16719329 TI - [Membrane skeleton: interaction of the plasma membrane with the cytoskeleton]. PMID- 16719330 TI - [Rho family: master regulators of cytoskeleton and cell migration]. PMID- 16719331 TI - [GEFs and gaps of Rho family GTPases]. PMID- 16719332 TI - [Activity of Rho-family G proteins as visualized with FRET-based probes]. PMID- 16719333 TI - [Application of soft lithography in the study of mechanotransduction]. PMID- 16719334 TI - [Motility of fibroblast cells and its regulatory mechanisms]. PMID- 16719335 TI - [Cytoskeleton and adhesion of the migratory neurons]. PMID- 16719336 TI - [Directional sensing and superoxide production in neutrophils]. PMID- 16719337 TI - [Signaling mechanism connecting axon guidance molecules and growth cone cytoskeleton]. PMID- 16719338 TI - [Dynamic cell migration during development]. PMID- 16719339 TI - [Regulatory network for activation of mitotic kinases]. PMID- 16719340 TI - [Cytokinesis: formation of the contractile ring during cell division]. PMID- 16719341 TI - [Dynamics of mitotic spindle]. PMID- 16719342 TI - [Actin organization regulated by lipids]. PMID- 16719343 TI - [Cell polarization: lessons from C. elegans asymmetric cell division]. PMID- 16719344 TI - [Regulation of epithelial cell polarity mediated by aPKC-PAR complex]. PMID- 16719345 TI - [Neuronal polarity]. PMID- 16719346 TI - [Loss of adhesive function of desmogleins in bullous diseases: pemphigus and impetigo]. PMID- 16719347 TI - [Breast cancer invasiveness: how does it emerge?]. PMID- 16719348 TI - Molecular beacons: Fluorescent probes for detection of endogenous mRNAs in living cells. AB - A novel approach for detecting nucleic acid in solution has been adopted for real time imaging of native mRNAs in living cells. This method utilizes hybridization probes, called "molecular beacons", that generate fluorescent signals only when they are hybridized to a complementary target sequence. Nuclease-resistant molecular beacons are designed to efficiently hybridize to accessible regions within RNAs and then be detected via fluorescence microscopy. The target regions chosen for probe binding are selected using two computer algorithms, mfold and OligoWalk, that predict the secondary structure of RNAs and help narrow down sequence stretches to which the probes should bind with high affinity in vivo. As an example, molecular beacons were designed against regions of oskar mRNA, microinjected into living Drosophila melanogaster oocytes and imaged via confocal microscopy. PMID- 16719349 TI - Second-harmonic imaging of collagen. AB - Molecules that have no center of symmetry are able to convert light to its second harmonic, at twice the frequency and half the wavelength. This only happens with any efficiency at very high light intensities such as are given by a pulsed laser, and because the efficiency of the process depends on the square of the intensity, it will be focal plane selective in exactly the same way as two-photon excitation of fluorescence. Because of its unusual molecular structure and its high degree of crystallinity, collagen is, by far, the strongest source of second harmonics in animal tissue. Because collagen is also the most important structural protein in the mammalian body, this provides a very useful imaging tool for studying its distribution. No energy is lost in second-harmonic imaging, so the image will not fade, and because it is at a shorter wavelength than can be excited by two-photon fluorescence, it can be separated easily from multiple fluorescent probes. It is already proving useful in imaging collagen with high sensitivity in various tissues, including cirrhotic liver, normal and carious teeth, and surgical repair of tendons. PMID- 16719350 TI - Visualizing calcium signaling in cells by digitized wide-field and confocal fluorescent microscopy. AB - Calcium (Ca2+) is a fundamentally important component of cellular signal transduction. Dynamic changes in the concentration of Ca2+ ([Ca2+]) in the cytoplasm and within organelles are tightly controlled and regulate a diverse array of biological activities, including fertilization, cell division, gene expression, cellular metabolism, protein biosynthesis, secretion, muscle contraction, intercellular communication, and cell death. Measurement of intracellular [Ca2+] is essential to understanding the role of Ca2+ and for defining the underlying regulatory mechanisms in any cellular process. A broad range of synthetic and biosynthetic fluorescent Ca2+ sensors are available that enable the visualization and quantification of subcellular spatio-temporal [Ca2+] gradients. This chapter describes the application of wide-field digitized video fluorescence microfluorometry and confocal microscopy to quantitatively image Ca2+ in cells with high temporal and spatial resolution. PMID- 16719351 TI - Multifluorescence labeling techniques and confocal laser scanning microscopy on lung tissue. AB - Lung tissue consists of more than 40 individual cell types that might interact to produce adverse pathologies. After injury, a number of signaling proteins expressed in various epithelial and other cell types have been linked to the advent of apoptosis, compensatory proliferation, and adaptation to stress. We describe here the use of immunochemistry and multifluorescence approaches using confocal laser scanning microscopy to define the signaling pathways (protein kinases C and mitogen-activated protein kinases) activated by asbestos fibers after inhalation. Using these approaches, we are able to localize signaling events in distinct cell types of the lung and determine their status in the cell cycle (resting or nonresting). Moreover, we are able to determine whether various signaling proteins colocalize in cells and the sites affected by asbestos fibers. PMID- 16719352 TI - Evaluation of confocal microscopy system performance. AB - The confocal laser scanning microscope (CLSM) has enormous potential in many biological fields. When tests are made to evaluate the performance of a CLSM, the usual subjective assessment is accomplished by using a histological test slide to create a "pretty picture." Without the use of functional tests, many of the machines could be working at suboptimal performance levels, delivering suboptimum performance and possibly misleading data. To replace the subjectivity in evaluating a confocal microscope, tests were derived or perfected that measure field illumination, lens clarity, laser power, laser stability, dichroic functionality, spectral registration, axial resolution, scanning stability, photomultiplier tube quality, overall machine stability, and system noise. These tests will help serve as a guide for other investigators to ensure that their machines are working correctly to provide data that are accurate with the necessary resolution, sensitivity, and precision. Utilization of this proposed testing approach will help eliminate the subjective nature of assessing the CLSM and allow different machines to be compared. These tests are essential if one is to make intensity measurements. PMID- 16719353 TI - Quantitative analysis of atherosclerotic lesion composition in mice. AB - Comparative quantitation has become an increasingly desirable tool in determining compositional differences of aortic plaque lesion in transgenically altered mice. To this end, methodology has been developed to identify lipid, cellularity, collagen, and elastin components using traditional bright-field microscopy, fluorescence, and polarized light microscopy, employing both confocal and wide field imaging systems. Subsequent imaging processing and analysis on the digitally captured images reveals differences in compositional components as influenced by diet, age and gender. This method can be expanded to employ a rich variety of histochemical and immunohistochemical staining protocols. PMID- 16719354 TI - Applications of microscopy to genetic therapy of cystic fibrosis and other human diseases. AB - Gene therapy has become an extremely important and active field of biomedical research. Microscopy is an integral component of this effort. This chapter presents an overview of imaging techniques used in our facility in support of cystic fibrosis gene therapy research. Instrumentation used in these studies includes light and confocal microscopy, transmission electron microscopy, and scanning electron microscopy. Techniques outlined include negative staining, cryo electron microscopy, three-dimentional reconstruction, enzyme cytochemistry, immunocytochemistry, and fluorescence imaging. PMID- 16719356 TI - Near-clinical applications of laser scanning cytometry. AB - Biological samples from human tissues are characterized by complexity and heterogeneity. The ability to make rapid, reliable, quantitative fluorochromatic measurements on clinical samples allows the development of new and practical assays that could influence diagnosis and treatment in a variety of clinical applications. Laser scanning cytometry (LSC) is a very versatile and adaptable technology that allows for the quantitative analysis of cell samples that are unsuitable for flow cytometry by virtue of their presentation and context. Crucially, it allows the direct visualization of cells and rare events and the correlation of imagery with fluorochromatic measurements. In this chapter, we describe early experiments in the study of cytotoxic drug uptake and resistance in human tumor cells and in the study of sputum cells from asthmatic patients, which harness the specific capabilities of LSC to practical clinical problems. PMID- 16719357 TI - Laser capture microdissection. AB - Laser capture microdissection (LCM) is a technique for isolating pure cell populations from a heterogeneous tissue section or cytological preparation via direct visualization of the cells. This technique is applicable to molecular profiling of diseased and disease-free tissue, permitting correlation of cellular molecular signatures with specific cell populations. DNA, RNA, or protein analysis can be performed with the microdissected tissue by any method with adequate sensitivity. The principle components of LCM technology are (1) visualization of the cells of interest via microscopy, (2) transfer of laser energy to a thermolabile polymer with formation of a polymer-cell composite, and (3) removal of the cells of interest from the heterogeneous tissue section. LCM is compatible with a variety of tissue types, cellular staining methods, and tissue-preservation protocols that allow microdissection of fresh or archival specimens. LCM platforms are available as a manual system (PixCell; Arcturus Bioscience) or as an automated system (AutoPix). PMID- 16719355 TI - Laser scanning cytometry: principles and applications. AB - The laser scanning cytometer (LSC) is the microscope-based cytofluorometer that offers a plethora of analytical capabilities. Multilaser-excited fluorescence emitted from individual cells is measured at several wavelength ranges, rapidly (up to 5000 cells/min), with high sensitivity and accuracy. The following applications of LSC are reviewed: (1) identification of cells that differ in degree of chromatin condensation (e.g., mitotic or apoptotic cells or lymphocytes vs granulocytes vs monocytes); (2) detection of translocation between cytoplasm vs nucleus or nucleoplasm vs nucleolus of regulatory molecules such as NF-kappaB, p53, or Bax; (3) semiautomatic scoring of micronuclei in mutagenicity assays; (4) analysis of fluorescence in situ hybridization; (5) enumeration and morphometry of nucleoli; (6) analysis of phenotype of progeny of individual cells in clonogenicity assay; (7) cell immunophenotyping; (8) visual examination, imaging, or sequential analysis of the cells measured earlier upon their relocation, using different probes; (9) in situ enzyme kinetics and other time-resolved processes; (10) analysis of tissue section architecture; (11) application for hypocellular samples (needle aspirate, spinal fluid, etc.); (12) other clinical applications. Advantages and limitations of LSC are discussed and compared with flow cytometry. PMID- 16719358 TI - Analysis of asbestos-induced gene expression changes in bronchiolar epithelial cells using laser capture microdissection and quantitative reverse transcriptase polymerase chain reaction. AB - Laser capture microdissection (LCM) enables the removal of discrete microstructures or cell types from properly prepared histological sections. Extraction of RNA from microdissected tissue followed by quantitative reverse transcriptase-polymerase chain (QRT-PCR) reaction permits the analysis of cell type or microstructure-specific gene expression changes that occur in response to various stimuli in the environment. In our lab, the combination of LCM and QRT PCR has proven very useful in the determination of the in vivo gene expression changes that occur in bronchiolar epithelium in response to inhalation of crocidolite asbestos. A detailed description of the preparation of cDNA from bronchiolar epithelial cells obtained by LCM is described in this work. PMID- 16719359 TI - New approaches to fluorescence in situ hybridization. AB - Fluorescence in situ hybridization (FISH) is a nonisotopic labeling and detection method that provides a direct way to determine the relative location or copy number of specific DNA sequences in nuclei or chromosomes. With recent advancements, this technique has found increased application in a number of research areas, including cytogenetics, prenatal diagnosis, cancer research and diagnosis, nuclear organization, gene loss and/or amplification, and gene mapping. The availability of different types of probe and the increasing number of FISH techniques has made it a widespread and diversely applied technology. Multicolor karyotyping by multicolor FISH and spectral karyotyping interphase FISH and comparative genomic hybridization allow genetic analysis of previously intractable targets. We present a brief overview of FISH technology and describe in detail methods of probe labeling and detection for different types of tissue sample, including microdissected nuclei from formalin-fixed paraffin-embedded tissue sections. PMID- 16719360 TI - Microarray image scanning. AB - Of the technologies available for measuring gene expression, microarrays using cDNA targets is one of the most common and well-developed high-throughput techniques. With this technique, the expression levels of thousands of genes are measured simultaneously. DNA probes are immobilized on solid surfaces, either membrane-based or chemically coated glass surfaces. On glass arrays, the probes are hybridized with fluorescent-labeled target samples. Fluorescence intensities, which reflect gene expression levels, are detected by imaging the array using a laser or white-light source and capturing the image using photomultiplier tube detection or a charge-coupled device camera. Different laser-based scanners are used in laboratories to scan microarray images. This chapter discusses the imaging process and the protocols being developed. PMID- 16719361 TI - Near-field scanning optical microscopy in cell biology and cytogenetics. AB - Light microscopy has proven to be one of the most versatile analytical tools in cell biology and cytogenetics. The growing spectrum of scientific knowledge demands a continuous improvement of the optical resolution of the instruments. In far-field light microscopy, the attainable resolution is dictated by the limit of diffraction, which, in practice, is about 250 nm for high-numerical-aperture objective lenses. Near-field scanning optical microscopy (NSOM) was the first technique that has overcome this limit up to about one order of magnitude. Typically, the resolution range below 100 nm is accessed for biological applications. Using appropriately designed scanning probes allows for obtaining an extremely small near-field light excitation volume (some tens of nanometers in diameter). Because of the reduction of background illumination, high contrast imaging becomes feasible for light transmission and fluorescence microscopy. The height of the scanning probe is controlled by atomic force interactions between the specimen surface and the probe tip. The control signal can be used for the production of a topographic (nonoptical) image that can be acquired simultaneously. In this chapter, the principle of NSOM is described with respect to biological applications. A brief overview of some requirements in biology and applications described in the literature are given. Practical advice is focused on instruments with aperture-type illumination probes. Preparation protocols focussing on NSOM of cell surfaces and chromosomes are presented. PMID- 16719362 TI - Porosome: the fusion pore revealed by multiple imaging modalities. AB - Secretion occurs in all cells of multicellular organisms and involves the delivery of secretory products packaged in membrane-bound vesicles to the cell exterior. Specialized cells for neurotransmission, enzyme secretion, or hormone release utilize a highly regulated secretory process. Secretory vesicles are transported to specific sites at the plasma membrane, where they dock and fuse to release their contents. Similar to other cellular processes, cell secretion is found to be highly regulated and a precisely orchestrated event. It has been demonstrated that membrane-bound secretory vesicles dock and fuse at porosomes, which are specialized supramolecular structures at the cell plasma membrane. Swelling of secretory vesicles results in a buildup of pressure, allowing expulsion of intravesicular contents. The extent of secretory vesicle swelling dictates the amount of intravesicular contents expelled during secretion. The discovery of the porosome, its isolation, its structure and dynamics at nanometer resolution and in real time, and its biochemical composition and functional reconstitution into artificial lipid membrane have been determined. The molecular mechanism of secretory vesicle fusion at the base of porosomes and vesicle swelling have also been resolved. These findings reveal the molecular machinery and mechanism of cell secretion. In this chapter, the discovery of the porosome, its isolation, its structure and dynamics at nanometer resolution and in real time, and its biochemical composition and functional reconstitution into artificial lipid membrane are discussed. PMID- 16719363 TI - Secretory vesicle swelling by atomic force microscopy. AB - The swelling of secretory vesicles has been implicated in exocytosis, but the underlying mechanism of vesicle swelling remained unknown. Earlier studies from our laboratory demonstrated the association of the alpha-subunit of heterotrimeric GTP-binding protein G(alphai3) with zymogen granule membrane and implicated its involvement in vesicle swelling. Mas7, an active mastoparan analog known to stimulate Gi proteins, was found to stimulate the GTPase activity of isolated zymogen granules and cause swelling. Increase in vesicle size in the presence of GTP, NaF, and Mas7 were irreversible and found to be KCl sensitive. However, Ca2+ had no effect on zymogen granule size. Taken together, these results indicated that zymogen granules, the membrane-bound secretory vesicles in exocrine pancreas, swell in response to GTP mediated by a G(alphai3) protein. Subsequently, our studies demonstrated that the water channel aquaporin-1 (AQP1) is also present at the zymogen granule membrane and participates in rapid GTP induced and G(alphai3)-mediated vesicular water gating and swelling. Isolated zymogen granules exhibit low basal water permeability. However, exposure of granules to GTP results in a marked potentiation of water entry. Treatment of zymogen granules with the known water channel inhibitor Hg2+ is accompanied by a reversible loss in both the basal and GTP-stimulable water entry and vesicle swelling. Introduction of AQP1-specific antibody raised against the carboxy terminal domain of AQP1 blocked GTP-stimulable swelling of vesicles. Our results demonstrate that AQPI associated at the zymogen granule membrane is involved in basal GTP-induced and G(alphai3)-mediated rapid gating of water into zymogen granules of the exocrine pancreas. PMID- 16719364 TI - Imaging and probing cell mechanical properties with the atomic force microscope. AB - This chapter describes the use of the atomic force microscope (AFM) to probe and map out regional variations in apparent elastic properties of living cells. The importance of mechanics in the field of cell biology is becoming more widely appreciated, and the AFM has unique advantages for cell mechanics applications. However, care must be taken in the acquisition, analysis, Band interpretation of AFM indentation data. To help make this powerful technique accessible to a broad range of investigators, detailed procedures are provided for all stages of the AFM experiment from sample preparation through data analysis and visualization. PMID- 16719365 TI - Reflection contrast microscopy: The bridge between light and electron microscopy. AB - Reflection contrast microscopy (RCM) is a light microscopic method to image cells at high definition and enhanced sensitivity compared to conventional bright-field microscopy. RCM images have very high contrast, which makes them easily applicable for digital image analysis. Because ultrathin sections are mostly used in this method, RCM also functions by bridging light with electron microscopy: the combination of ultrastructural with histochemical studies. RCM can also replace electron microscopy for rapid and simple screening of large quantities of samples for immunocytochemical staining. Special attention is paid to small biological objects, which have to be processed for RCM. If you encounter the limits of brightfield microscopy, in resolution, sensitivity or handling of the specimen, RCM will be a feasible option. Reflection contrast microscopy methods use only slightly adjusted electron microscopy methods for specimen preparation. Therefore, many familiar techniques for ultrathin specimen preparation can be applied. It is essential that only refractive index differences exist in those areas that are of interest and that the further specimen is as optically homogenic as possible, with a refractive index as close to that of glass as possible. Therefore, plastic embedding is recommended. PMID- 16719366 TI - Three-dimensional analysis of single particles by electron microscopy: sample preparation and data acquisition. AB - Electron microscopy of single particles has recently become a very popular field in both biological and material sciences. It might be difficult for a novice researcher new to this field to know how to start tackling a new project. This chapter is designed to serve as a guideline for anyone starting a new project to determine a three-dimensional structure using single-particle techniques. The chapter describes the basic techniques necessary to prepare the samples and acquire the data to calculate a three-dimensional reconstruction in easy-to understand, step-by-step instructions. It starts with the basic preparation of support films and the usage of a variety of staining techniques needed to assess the quality of the sample and the viability of the project. It ends with a detailed description of vitreous ice preparations designed to acquire high resolution structural information. Guidelines and tips are given on how to record the best images with an electron microscope. Although this chapter is geared to researchers new to the field, experts might find it not only useful as a reference but also valuable because of the number of practical tips included. PMID- 16719367 TI - Three-dimensional reconstruction of single particles in electron microscopy image processing. AB - Three-dimensional electron microscopy of single macromolecular assemblies has made large strides forward over the last decade. A large number of image processing techniques have been developed and many have found general distribution. For the proper usage of the wide range of available techniques, a clear concept of all processing steps is essential. This chapter provides step-by step instruction for the three-dimensional reconstruction of an unknown macromolecule. Where possible, the limitations of the techniques are explained. The chapter attempts to be sufficiently general such so as not to adhere to a single image processing system. Described are alignment techniques for two and three dimensions, classification procedures, and the usage of three-dimensional reconstruction algorithms. PMID- 16719368 TI - A new microbiopsy system enables rapid preparation of tissue for high-pressure freezing. AB - A microbiopsy system was developed to overcome long sampling times for tissues before they are cryo-fixed by high-pressure freezing. A commercially available biopsy gun was adapted to the needs of small-organ excisions, and biopsy needles were modified to allow small samples (0.6 mm x 1.2 mm x 0.3 mm) to be taken. Specimen platelets with a central slot of the same dimensions as the biopsy are used. A self-made transfer device (in the meantime optimized by Leica Microsystems [Vienna, Austria]) coordinates the transfer of the excised sample from the biopsy needle into the platelet slot and the subsequent loading in a specimen holder, which is then introduced into a high-pressure freezer (Leica EM PACT; Leica Microsystems, Vienna, Austria). Thirty seconds preparation time is needed from excision until high-pressure freezing. Brain, liver, kidney and muscle excisions of anesthetised rats are shown to be well frozen. PMID- 16719369 TI - Cytochrome P450 nomenclature, 2004. AB - Aspects of cytochrome P450 (CYP) nomenclature are addressed. The rules for naming a P450 are outlined, though individuals should not name their own genes. The nomenclature is presented as a unifying principle to enhance communication across disciplines. Because of the historical nature of gene sequencing, sometimes names have to be changed, but this is kept to a bare minimum to avoid confusion in the literature. CYP names have now reached four digits owing to proliferation of CYP families in the fungi and lower eukaryotes. For example, CYP5034A1 is from Ustilago maydis. P450 sequence motifs are described that are useful in making global alignments. CYP clans are defined as clusters of CYP families. The clan names are useful in describing higher-order evolution of the gene superfamily. The nomenclature of orthologs and pseudogenes is also discussed. PMID- 16719370 TI - Spectral analyses of cytochromes P450. AB - UV/Vis spectroscopy is the major means of identifying intact holocytochrome P450. The carbon monoxide complex of the intact ferrous hemoprotein exhibits a characteristic spectrum between 448 and 452 nm, considerably distinct from the usual Soret absorption peaks of hemoproteins. Methods are described for identification and quantitation of cytochrome P450 (CYP) in membranes, in tissue homogenates, and in purified form, using difference spectroscopy and absolute spectroscopy. CYP are b-type cytochromes, containing protoporphyrin IX as the prosthetic group. Methods are also provided, using alkali and pyridine, for quantitation of the hemoprotein by this prosthetic group. In its oxidized, or ferric state, CYP exists as an equilibrium mixture of high- and low-spin configurations, each with distinctive UV/Vis absorption peaks. Substrate binding causes shifts in the spin equilibrium, and methods are shown for using these shifts for quantitation of substrate binding to CYP. PMID- 16719371 TI - Establishment of functional human cytochrome P450 monooxygenase systems in Escherichia coli. AB - Cytochromes P450 (CYP) have been expressed in a variety of systems such as mammalian cells, yeast, and bacteria. The bacterial system is technically the least demanding and provides large amounts of catalytically active P450s for metabolic and structural studies relating to preclinical drug development. This chapter provides a detailed technical description of the processes that allow the coexpression of various CYP isoforms together with CYP reductase in Escherichia coli and gives some examples of the results that can be achieved for the expression of human P450s. PMID- 16719372 TI - Purification of cytochromes P450: products of bacterial recombinant expression systems. AB - A general procedure for the solubilization of cytochrome P450 (P450) from bacterial membranes specifically for a human P450 expressed heterologously in the host Escherichia coli is described. The example involves the use of a P450 (3A4) with a C-terminal oligohistidine tag and includes sequential DEAE and metal affinity chromatography. PMID- 16719373 TI - Expression of recombinant flavin-containing monooxygenases in a baculovirus/insect cell system. AB - The baculovirus/insect cell heterologous expression system provides an important tool for investigating the catalytic activity of individual drug-metabolizing enzymes toward a particular substrate. In this chapter we describe a baculovirus/insect cell system that we have used for the expression of human and mouse flavin-containing monooxygenases. Methods are described for the generation of recombinant baculoviral DNAs, via both site-specific transposition in Escherichia coli and site-specific recombination in vitro; adaptation of Spodopterafrugiperda (Sf) 9 cells to shaking culture and to serum-free medium; cryopreservation and transfection of Sf9 cells; amplification of baculovirus and determination of viral titer; analysis of baculoviral DNA; and expression and analysis of recombinant proteins. PMID- 16719374 TI - Cytochrome P450 reconstitution systems. AB - Human liver microsomes contain multiple forms of cytochrome P450 (CYP or P450) that catalyze oxidation of a number of xenobiotic and endobiotic chemicals. Individual P450 forms have unique, but overlapping, substrate specificities. It is necessary to determine which P450s play more important roles in the oxidation of these chemicals. A good way of studying the roles of P450s in the metabolism of these chemicals is to reconstitute the activities by mixing purified P450s and nicotinamide adenine dinucleotide phosphate-cytochrome P450 reductase in the membranes of phospholipid vesicles. However, our studies have suggested that the conditions for reconstitution of activities vary depending on the P450 enzymes used. For example, some reactions catalyzed by P450s require cytochrome-b5 and a particular phospholipid environment for exerting their full catalytic activities. In this chapter, we describe optimal conditions that have been determined in our laboratories for the reconstitution of drug oxidation activities catalyzed by purified human CYP1A2, 2C9, 2E1, and 3A4. PMID- 16719375 TI - Catalytic assays for human cytochrome P450: an introduction. AB - Cytochrome P450 (P450) is a superfamily of individual monooxygenase enzymes that metabolize structurally diverse xenochemicals, including many clinically useful drugs and foreign chemicals widespread in the environment. P450 substrates that can be used to selectively monitor individual P450 enzymes or P450 subfamilies have been identified through studies using P450 enzyme-selective inhibitory antibodies and chemical inhibitors in conjunction with experiments utilizing individual cDNA-expressed P450 enzymes. This chapter describes P450 form selective substrates that can be used to monitor the activities of human P450 enzymes CYP1A, CYP1B1, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A, CYP4A11, and CYP7A1. Cautions that need to be exercised when using these substrates to probe for individual P450 activities in human liver and other tissues are discussed. PMID- 16719376 TI - Enzymatic analysis of cDNA-expressed human CYP1A1, CYP1A2, and CYP1B1 with 7 ethoxyresorufin as substrate. AB - Cytochrome P450 (P450) enzymes belonging to the CYP1 family are highly inducible by polycyclic aromatic hydrocarbons and other environmental chemicals and play a major role in the metabolism of many foreign chemicals and endogenous substances. We describe a spectrofluorometric method for determining 7-ethoxyresorufin O dealkylation catalyzed by CYP1A1, CYP1A2, and CYPB1. The formation of the enzymatic product, resorufin, is monitored continuously by fluorescence using an excitation wavelength of 530 nm and an emission wavelength of 580 nm. This method can be applied to assay P450-catalyzed formation of resorufin from other alkoxyresorufins, such as 7-methoxyresorufin, 7-benzyloxyresorufin, and 7 pentoxyresorufin. It can also be used to assay 7-ethoxyresorufin O-dealkylation activity in isolated hepatocytes and cultured cells that express this P450 activity. PMID- 16719377 TI - Spectrofluorometric analysis of CYP2A6-catalyzed coumarin 7-hydroxylation. AB - A spectrofluorometric method is described for the determination of CYP2A6 catalyzed coumarin 7-hydroxylation. Following acidification of the reaction mixture, the enzymatic product, 7-hydroxycoumarin, is recovered by a double extraction procedure and assayed using an excitation wavelength of 370 nm and an emission wavelength of 450 nm. This assay is applicable to enzymatic studies of cDNA-expressed CYP2A6 and can be used to monitor coumarin 7-hydroxylation activity in microsomes prepared from liver and other tissues and in isolated hepatocytes and cultured cells that express this cytochrome P450 activity. PMID- 16719378 TI - Determination of CYP2B6 component of 7-ethoxy-4-trifluoromethylcoumarin O deethylation activity in human liver microsomes. AB - The cytochrome P450 enzyme CYP2B6 plays an important role in the metabolism of structurally diverse drugs, including the anticancer drug cyclophosphamide, and may be an important determinant of clinical responses to these agents. A spectrofluorometric method is described for the determination of CYP2B6-catalyzed 7-ethoxy-4-trifluoromethylcoumarin O-deethylation activity in human liver microsomes. The specificity of this method for CYP2B6 is increased by the use of inhibitory antibodies to CYP1A2, CYP2C, and CYP2E1, which block the contributions of these higher-K(m) enzymes to human liver microsomal metabolism of 7-ethoxy-4 trifluoromethylcoumarin. The enzymatic product, 7-hydroxy-4 trifluoromethylcoumarin, is monitored by fluorescence using an excitation wavelength of 410 nm and an emission wavelength of 510 nm. This approach can be modified to assay the catalytic activity of cDNA-expressed CYP2B6. PMID- 16719379 TI - High-performance liquid chromatography analysis of CYP2C8-catalyzed paclitaxel 6alpha-hydroxylation. AB - Paclitaxel (Taxol) is a naturally occurring member of the taxane family of antitumor drugs, which act by stabilizing microtubules. Paclitaxel is inactivated in human liver by a cytochrome P450 (P450)-catalyzed 6alpha-hydroxylation reaction. A reverse-phase, high-performance liquid chromatographic assay is described for the analysis of paclitaxel 6alpha-hydroxylation catalyzed by human liver microsomes or cDNA-expressed P450 enzyme CYP2C8. Analytical separations are achieved using a C18 column with a linear gradient of 10-100% methanol, with detection at 230 nm. This method is applicable to enzymatic studies for determination of CYP2C8-catalyzed paclitaxel 6alpha-hydroxylation activity. PMID- 16719380 TI - Determination of CYP2C9-catalyzed diclofenac 4'-hydroxylation by high-performance liquid chromatography. AB - A reverse-phase, high-performance liquid chromatography method is described for quantification of diclofenac 4'-hydroxylation catalyzed by human liver microsomes or cDNA-expressed CYP2C9. Analytical separation is achieved using a C18 column developed with a gradient of 30% acetonitrile and 2 mM perchloric acid in water to 100% methanol, with detection at 280 nm. This method is applicable to enzymatic studies for determination of CYP2C9-catalyzed diclofenac 4' hydroxylation activity. PMID- 16719381 TI - CYP2C19-mediated (S)-mephenytoin 4'-hydroxylation assayed by high-performance liquid chromatography with radiometric detection. AB - A reverse-phase, high-performance liquid chromatography method is described for the quantification of 4'-hydroxymephenytoin formed enzymatically from 14C-labeled (S)-mephenytoin following incubation with cDNA-expressed CYP2C19 or human liver microsomes. Analytical separation is achieved using a C18 column developed with a gradient from 10 to 100% methanol, with detection using a scintillation detector. This method is applicable to enzymatic studies for determination of CYP2C19 catalyzed (S)-mephenytoin 4'-hydroxylation activity. PMID- 16719382 TI - CYP2D6-dependent bufuralol 1'-hydroxylation assayed by reverse-phase ion-pair high-performance liquid chromatography with fluorescence detection. AB - A reverse-phase, high-performance liquid chromatography method is described for the quantification of 1'-hydroxybufuralol formed enzymatically by the incubation of bufuralol with cDNA-expressed CYP2D6 or human liver microsomes. Analytical separation is achieved using a C18 column and a mobile phase consisting of 30% acetonitrile and 2 mM perchloric acid, with detection by fluorescence using an excitation wavelength of 252 nm and an emission wavelength of 302 nm. This method is applicable to enzymatic studies for determination of CYP2D6-catalyzed bufuralol 1'-hydroxylation activity. PMID- 16719383 TI - Spectrophotometric analysis of human CYP2E1-catalyzed p-nitrophenol hydroxylation. AB - The cytochrome P450 enzyme CYP2E1 catalyzes the oxidative metabolism of many solvents and other small organic molecules. A spectrophotometric method is described for determination of CYP2E1 activity by monitoring the formation of p nitrocatechol from p-nitrophenol by cDNA-expressed CYP2E1 or isolated liver microsomes. The enzymatic product, p-nitrocatechol, is assayed at 535 nm after acidification of the reaction mixture with trichloroacetic acid followed by neutralization using 2 M NaOH. This method is applicable to enzymatic studies for determination of P450-catalyzed p-nitrophenol hydroxylation activity. PMID- 16719384 TI - Thin-layer chromatography analysis of human CYP3A-catalyzed testosterone 6beta hydroxylation. AB - Testosterone and other steroid hormones have been studied as prototypic examples of endogenous substrates for hepatic cytochrome P450 (P450) enzymes. CYP3A enzymes from various species, including human, metabolize testosterone by a 6beta hydroxylation reaction, which is unique to this P450 subfamily. A thin-layer chromatographic method is described for the determination of 6beta hydroxytestosterone formed enzymatically by incubation of [14C]-testosterone with cDNA-expressed CYP3A enzymes or liver microsomes. 14C-labeled enzymatic products are applied to silica gel thin-layer plates, which are developed sequentially with methylene chloride:acetone (80:20) followed by chloroform, ethyl acetate, and absolute ethanol (80:20:14). Metabolite quantification is performed by autoradiography and liquid scintillation counting. This method is applicable to enzymatic studies for the determination of CYP3A-dependent testosterone 6beta hydroxylation activity in both human and animal liver microsomes. PMID- 16719385 TI - Determination of CYP4A11-catalyzed lauric acid 12-hydroxylation by high performance liquid chromatography with radiometric detection. AB - Lauric acid serves as an endogenous substrate for the cytochrome P450 enzyme CYP4A11. A reverse-phase, high-performance liquid chromatography method is described for the quantification of 12-hydroxylauric acid formed enzymatically by incubation of 14C-labeled lauric acid with cDNA-expressed CYP4A11 or human liver microsomes. Analytical separation is achieved using a C18 column and a gradient of 30% acetonitrile and 2 mM perchloric acid to 100% methanol, using a detection scintillation counter. This method is applicable to enzymatic studies for determination of lauric acid 12-hydroxylation activity. PMID- 16719386 TI - An isocratic high-performance liquid chromatography assay for CYP7A1-catalyzed cholesterol 7alpha-hydroxylation. AB - A normal-phase, isocratic high-performance liquid chromatography assay is described for cholesterol 7alpha-hydroxylation catalyzed by CYP7A1, which corresponds to the first and rate-limiting step in the conversion of cholesterol into bile acids. This method is based on the conversion of the primary cytochrome P450 metabolite, 7alpha-hydroxycholesterol, into 7alpha-hydroxy-4-cholesten-3-one in a reaction catalyzed by exogenous cholesterol oxidase, followed by chromatographic separation with monitoring at 254 nm. This technique is applicable to enzymatic studies for determination of cholesterol 7alpha hydroxylation activity catalyzed by cDNA-expressed CYP7A1 and animal or human liver microsomes. PMID- 16719388 TI - Benzydamine N-oxygenation as a measure of flavin-containing monooxygenase activity. AB - Benzydamine is a nonsteroidal anti-inflammatory drug that undergoes flavin containing monooxygenase (FMO)-dependent metabolism to a stable N-oxide. This metabolite can be quantified with high specificity and sensitivity by using a simple reverse-phase high-performance liquid chromatography (HPLC) assay with fluorescence detection. Studies with recombinant FMO enzymes demonstrate that FMOI and FMO3 are the primary catalysts of benzydamine N-oxygenation, with minimal contributions from cytochrome P450 enzymes. Investigations conducted with human liver microsomes confirm that FMO3, in large part, is responsible for benzydamine N-oxide formation in this tissue. These features render benzydamine a useful in vitro probe for FMO activity in a wide range of tissues and cell types. In addition, benzydamine appears to be a suitable in vivo probe for human liver FMO3. This chapter provides a detailed account of the experimental protocol for determining rates of formation of benzydamine N-oxide by FMO-containing enzyme fractions. PMID- 16719387 TI - Use of 7-ethoxycoumarin to monitor multiple enzymes in the human CYP1, CYP2, and CYP3 families. AB - 7-Ethoxycoumarin is metabolized by many cytochrome P450 enzymes active in foreign compound metabolism and has been used as a prototypic substrate to monitor P450 (P450) activity in both hepatic and extrahepatic tissues. A spectrofluorometric method is described for determination of P450-catalyzed 7-ethoxycoumarin O deethylation. Following acidification of the incubation mixture, the enzymatic product, 7-hydroxycoumarin, is recovered by a double-extraction procedure and measured at an excitation wavelength of 370 nm and an emission wavelength of 450 nm. This method is applicable to enzymatic studies to determine the catalytic activity of cDNA-expressed human enzymes in the CYP1, CYP2, and CYP3 families, and 7-ethoxycoumarin O-deethylation activity in microsomes isolated from liver and other tissues. PMID- 16719389 TI - Prochiral sulfoxidation as a probe for flavin-containing monooxygenases. AB - Asymmetric aryl alkyl sulfides (R-S-R') are metabolized by flavin-containing monooxygenase (FMO) and cytochrome P450 enzymes to enantiomerically enriched sulfoxide products (R-SO-R') that are readily analyzed with a host of commercially available chiral stationary phases. Prochiral sulfoxidation of probe compounds based on p-tolyl methyl sulfide is a particularly useful method for discriminating among FMO1, FMO3, and FMO5, because the stereochemistry of the resulting products is isoform dependent, but apparently species independent. If studies are performed with crude tissue microsomal preparations, the cytochrome P450 component must be quenched to unmask catalysis specifically by the FMO component of the tissue. This chapter details experimental protocols for stereochemical analysis of sulfoxides generated from methyl, ethyl, n-propyl, and n-butyl p-tolyl sulfide by purified FMO isoforms and tissue microsomal preparations. PMID- 16719390 TI - Targeting antipeptide antibodies toward cytochrome P450 enzymes. AB - An approach to raising antibodies is described that can be applied to the majority of cytochrome P450 (P450) enzymes. Its application is limited only by the availability of suitable protein sequence information. The method is based on immunizing animals with synthetic peptides that mimic small regions of target proteins. In practice, this has proved to be a relatively simple, rapid, and effective method of producing antibodies. The antibodies are particularly suited to immunoblotting (Western blotting) and immunocytochemistry. Unlike antibodies produced by other techniques, the epitopes of these antibodies are predetermined, allowing them to be directed toward specific regions of P450. Detailed practical information is described on the selection of regions to target, the synthesis and conjugation of peptides to carrier protein, immunization, and the assessment of the resultant antisera. The approach is illustrated using examples of antibodies targeted against rat and human P450. PMID- 16719391 TI - The human cytochrome P450 Allele Nomenclature Committee Web site: submission criteria, procedures, and objectives. AB - Interindividual variability in xenobiotic metabolism and drug response is extensive. Genetic factors are predicted to account for 15-30% of this variability in general, but for certain drugs the genetic factor is the major determinant for outcome of drug therapy. Of particular importance for drug metabolism, drug response, and adverse drug reactions are the cytochrome P450 (CYP) enzymes, many of which are polymorphic. An essential basis for research and applications regarding interindividual variability in xenobiotic metabolism and toxicity by polymorphic CYPs is to have a common nomenclature for genetic variants and a system that allows researchers to be rapidly updated within the field. Since 1999 this has been achieved by the operation of the Human Cytochrome P450 Allele Nomenclature Committee Web site (www.imm.ki.se/CYPalleles/), where novel allelic variants are published after peer review. Currently, this Web site covers the nomenclature for polymorphic alleles of 22 CYP isoforms including more than 200 functionally different variants. Each CYP has its own Web page, which lists the alleles with their nucleotide changes, their functional consequences, and links to publications where the allele has been identified and characterized. The CYP allele Web site offers a rapid on-line publication of new alleles, provides an overview of peer-reviewed data, and serves as a form of quality control on research on new alleles. PMID- 16719392 TI - Genotyping for cytochrome P450 polymorphisms. AB - Protocols for the extraction of DNA from human blood and for genotyping for a number of common cytochrome P450 polymorphisms using either polymerase chain reaction (PCR)-restriction fragment length polymorphism or PCR-single-strand conformational polymorphism (SSCP) analysis are described. Rapid high-throughput techniques are also available for analyses of this type, but they require access to specialized equipment and are not considered here. General guidelines for performing amplification using PCR are described together with electrophoresis protocols for analysis of restriction digests of PCR products with agarose and polyacrylamide gels including the use of polyacrylamide-based gels for SSCP analysis. Protocols for the following specific isoforms and alleles are also provided: CYP1A1 (*2B and *4 alleles), CYP2C8 (*3 and *4 alleles), CYP2C9 (*2, *3, and *11 alleles), CYP2C19 (*2 and *3 alleles), CYP2D6 (*3, *4, *5, and *6 alleles), CYP2E1 (*5A, *5B, and *6 alleles), and CYP3A5 (*3 allele). PMID- 16719393 TI - Hepatocyte cultures in drug metabolism and toxicological research and testing. AB - When a new chemical entity is tested for its safety for humans and their environment, in vivo experiments on living animals are usually conducted. However, in the early preclinical stage of drug development, in vitro techniques, and more specifically hepatocyte-based in vitro models, are currently being applied. A major problem, however, related to the use of hepatocytes and their cultures is their limited viability, which is associated with the loss of phase I and phase II biotransformation capacity. Therefore, in order to keep the hepatocytes functional for a longer period, the in vivo microenvironment is mimicked in vitro as closely as possible through the addition of soluble medium components, coculture with helper cells, or culture on an extracellular matrix (sandwich culture). We discuss the advantages and disadvantages of current short- and long-term hepatocyte culture systems are discussed as well as their proper use in toxicological research and testing. PMID- 16719394 TI - Isolation of rat hepatocytes. AB - In vitro models, based on liver cells or tissues, are indispensable in the early preclinical phase of drug development. An important breakthrough in establishing cell models has been the successful high-yield preparation of intact hepatocytes. In this chapter, the practical aspects of the two-step collagenase perfusion method, modified from the original procedure of Seglen, are outlined. Although applicable to the liver of various species, including human, the practical aspects of the method are explained here for rat liver. Critical parameters for the successful isolation of primary rat hepatocytes are highlighted and a troubleshooting guide is provided. In addition, a new development based on the inhibition of histone deacetylase activity is presented. This approach allows inhibition of cell-cycle reentry during hepatocyte isolation, a process known to underlie the dedifferentiation process of cultured hepatocytes. PMID- 16719395 TI - Rat hepatocyte cultures: conventional monolayer and cocultures with rat liver epithelial cells. AB - Primary cultures of hepatocytes are useful tools for both short- and long-term pharmacotoxicological research. Under conventional conditions, isolated hepatocytes form a monolayer and survive for about 1 wk but lose some liver specific functions, including xenobiotic biotransformation. In comparison with the conventional monolayer culture model, cocultures with rat liver epithelial cells (RLECs) have an extended lifespan and better maintain their drug metabolizing capacity, owing to the presence of cell-cell interactions. In this chapter, techniques for setting up conventional monolayer cultures and cocultures of hepatocytes with RLECs (including isolation, culture, and cryopreservation of RLECs) are described in detail. In addition, comments derived from our own experience are given for successfully culturing primary hepatocytes. PMID- 16719396 TI - Rat hepatocyte cultures: collagen gel sandwich and immobilization cultures. AB - Mimicking the in vivo microenvironment is one of the current strategies to maintain liver-specific functionality in primary cultured hepatocytes for long periods. Freshly isolated hepatocytes entrapped in collagen gel type I (collagen gel immobilization culture) or sandwiched between two layers of hydrated collagen type I (collagen gel sandwich culture) are known to display liver-specific functions (e.g., biotransformation capacity) for more than 6 wk. We describe how to set up both types of organotypical hepatocyte culture systems. Besides a detailed protocol, we give some practical tips, taken from our own experience with long-term hepatocyte culture. PMID- 16719397 TI - Hepatocytes in suspension. AB - Isolated hepatocytes are a physiologically relevant in vitro model exhibiting intact subcellular organelles, xenobiotic transport, and integrated phase I and phase II biotransformation. They represent the "gold standard" for investigating xenobiotic biotransformation and metabolic bioactivation. When used in suspension, they provide an easy-to-handle and relatively cheap in vitro system that can be used for up to 4 h. The use of animal- and human-derived hepatocytes allows interspecies comparisons of metabolic properties. In contrast with microsomes, which are easily prepared from human liver tissue and can be stored in liquid nitrogen with minimal loss of functionality, cryopreservation of isolated human hepatocytes has been shown to be more difficult: after thawing losses of cell viability and biotransformation capacity occur. We provide general recommendations for the appropriate use of hepatocytes in suspension for pharmaco toxicological studies. We also provide protocols for the cryopreservation of freshly isolated hepatocytes and their handling on thawing. PMID- 16719398 TI - Isolation of rat bone marrow stem cells. AB - Stem cell research has become an important field of study for molecular, cellular, and clinical biology as well as pharmaco-toxicology. Indeed, stem cells have a strong proliferative and unlimited self-renewal potential and are multipotent. In vivo as well as in vitro studies have confirmed the differentiation of adult bone marrow stem cells into muscle cells, adipocytes, cardiomyocytes, neuroectodermal cells, osteoblasts, chondroblasts, and so on. Recently, it has been shown that, under appropriate culture conditions, adult bone marrow stem cells may also differentiate into hepatocyte-like cells. Because of their extensive proliferative capacity and pluripotency, adult bone marrow stem cells could serve in the future as an unlimited source of hepatocytes for pharmaco-toxicological research and testing. We describe a protocol for isolation of mononuclear cells from adult rat bone marrow. PMID- 16719399 TI - Transfection of primary cultures of rat hepatocytes. AB - Five different transfection reagents-calcium phosphate, TransFast Transfection Reagent, Superfect Transfection Reagent, Effectene Transfection Reagent, and Tfx 20--were compared for their ability to effectively transfect primary cultures of male rat hepatocytes. Hepatocytes were isolated by the collagenase perfusion method and then cultured on Matrigel-coated plates for 24 h before transfection. The cells were transfected with either pGL3-Control or pGL3-Basic plasmids. The efficiency of transfection of each reagent was monitored using the dual luciferase reporter gene assay system. Superfect Transfection Reagent, Effectene Transfection Reagent and Tfx-20 were the most effective for the transfection of primary hepatocytes and gave comparable transfection efficiencies. Calcium phosphate was found to be the least effective transfection reagent and gave the most variable transfection results. Tfx-20 gave the least variable transfection results when different hepatocyte preparations were compared. PMID- 16719400 TI - Human hepatocyte culture. AB - Primary culture of human hepatocytes is an in vitro model widely used to investigate numerous aspects of liver physiology and pathology. The technique used to isolate human hepatocytes is based on two-step collagenase perfusion. Originally performed in situ for obtaining hepatocytes from the adult rat, this technique has been adapted to the ex vivo treatment of human liver from organ donors or from lobectomy resection for medical purposes. This chapter describes experimental protocols for the isolation of hepatocytes from human liver tissue and for the preparation of short- and long-term cultures in which cells retain a differentiated phenotype for at least 1 mo. The various aspects emphasized here include the conditions for obtaining tissue, quality control of tissue for efficient perfusion, collagenase perfusion parameters, solutions for perfusion and culture media, cell substrate, cell plating, specific equipment, and safety conditions. PMID- 16719401 TI - Determination of cellular localization of expression of flavin-containing monooxygenase genes in mouse tissues by in situ hybridization. AB - Methods are described for the cellular localization of expression of flavin containing monooxygenase (FMO) genes in various mouse tissues by in situ hybridization. These include the production of digoxigenin (DIG)-labeled antisense and sense RNA probes by transcription from FMO cDNA templates, the preparation of paraffin wax-embedded and cryostat tissue sections, the hybridization of RNA probes to tissue sections, and the specific detection of hybridized probes using an antibody to DIG. PMID- 16719402 TI - Deletion of genes from the mouse genome using Cre/loxP technology. AB - The steps required to delete genes from the mouse genome are illustrated by showing how a cluster of three flavin-containing monooxygenase (Fmo) genes (Fmol, Fmo2, and Fmo4) were deleted from mouse chromosome 1. Such large deletions are accomplished using loxP/Cre recombinase technology. Genomic clones corresponding to the genes to be deleted are first isolated, and then appropriate genomic fragments are cloned into vectors containing a loxP site. This produces targeting vectors, which are electroporated into mouse embryonic stem (ES) cells to allow a homologous recombination event to take place between the mouse genomic fragment, present within the vector, and the homologous sequences in the ES cell genome. Screening of ES cells for recombinants in which loxP sites have been inserted on either side of the gene cluster to be deleted is described. Recombination by Cre recombinase to produce ES cell lines carrying the deletion on chromosome 1 is also described. PMID- 16719403 TI - Characterization of targeted mouse embryonic stem cell chromosomes: karyotyping and fluorescence in situ hybridization of metaphase spreads. AB - The manipulation of genes in mouse embryonic stem (ES) cells can result in chromosome abnormalities. This chapter describes methods for karyotyping of the manipulated ES cell line before injection into blastocysts and the use of fluorescence in situ hybridization to confirm the deletion of a targeted gene. The method is illustrated by describing how an ES cell line targeted for the deletion of Fmo genes was characterized. PMID- 16719404 TI - Microinjection of targeted embryonic stem cells and establishment of knockout mouse lines for Fmo genes. AB - Methods are described for the injection of mouse embryonic stem cells, in which Fmo genes have been targeted to disrupt gene function, into 3.5-d-old blastocysts and the implantation of these into foster mothers. Successful injection and implantation of blastocysts will produce mice of mixed coat color (the chimera). Also described are methods to establish the success of blastocyst injection and implantation of germ-line transmission of the knockout (KO) mutation. Breeding strategies to produce congenic and isogenic KO mouse lines are outlined. Simple methods for the isolation of tail DNA, the tagging of mice, and record keeping of the line are also given. PMID- 16719405 TI - Use of reporter genes to measure xenobiotic-mediated activation of CYP gene transcription. AB - Transcriptional activation of CYP gene expression by xenobiotics may have fundamental effects on body physiology. It may result in the altered pharmacokinetics of other chemicals in the body, both xenobiotic and endogenous substrates, potentially altering their effects. This may often result in no observable clinical effect, but in a significant number of cases these interactions lead to altered physiology or failure of a therapeutic drug. It is therefore important to be able to screen novel chemical entities for their ability to activate CYP gene expression. In addition, through mechanistic studies of how such transcriptional activation occurs, the ability to predict and avoid such potential interactions is improved. Reporter gene assays provide a simple, high-throughput methodology for examining the transcriptional activation of CYP gene expression by xenobiotics. They are suitable for use in screening as well as mechanistic studies and are of use in both the drug discovery/development and research arenas. PMID- 16719406 TI - Two novel cell specific receptor proteins, CRLR and CD 117 in human glial tumors. AB - OBJECTIVE: CRLR (calcitonin receptor-like receptor) and CD 117, the gene product of c-kit have been shown to be expressed in cells of glial tumors, especially in those with higher malignancy. Here we report the distribution of these peptides in various cellular compartments within those tumors. MATERIAL: Both receptor proteins have been investigated in 95 glial tumor biopsies of different grades. METHODS: Both proteins were visualized by immunohistochemistry with antibodies either commercially available or raised for this purpose. RESULTS: Both receptor peptides can be identified in or around tumor blood vessels. CRLR occurs in some endothelial cells, especially in the microvascular proliferations of glioblastoma multiforme, whereas CD 117 preferentially occurs in cells of the thickened vascular wall within cells of pericyte or fibroblast morphology. Both antigens are found in addition in few neoplastic cells of overt astrocyte morphology. CONCLUSIONS: The occurrence of identical antigens in glial tumor blood vessels and in neighboring tumor cells underlines the common origin of "mesenchymal" and "neuroepithelial" components of such (malignant) glial neoplasms. PMID- 16719407 TI - Fatal Epstein-Barr virus meningitis: an autopsy report. AB - A 3-year-old Asian female presented with fever for 1 week and neck swelling for 1 day. Serology revealed a recent Epstein-Barr virus (EBV) infection. Late on the evening of admission, she developed confusion and would not follow commands. A CT scan showed diffuse cerebral edema and a cerebral flow scan demonstrated no blood flow to the brain. She was declared brain dead and expired on the following day. At autopsy, the brain weighted 1175 grams and grossly showed significant edema. Microscopically, the entire neuraxis revealed extensive leptomeningeal infiltrate of mainly CD8+ T lymphocytes, the majority of which expressed activated markers, HLA-DR and/or CD45RO, and monocytes/macrophages with intermixed numerous apoptotic/karyorrhectic nuclear fragments. These nuclear fragments were considered to be due to apoptosis of the expanded population of CD8+ T lymphocytes. Focal venulitis was noted. EBV-encoded small nuclear RNA in situ hybridization revealed positivity in the occasional lymphocytes. Interestingly, most intraparenchymal as well as leptomeningeal vascular endothelium showed HLA DR immunoreactivity. This finding has been reported primarily in the acute inflammatory/demyelinating conditions, not in the viral meningitis/meningoencephalitis, and was thought to be related to cytokines due to widespread inflammation in our case. Massive edema secondary to severe EBV meningitis can be fatal. PMID- 16719409 TI - Short rib-polydactyly syndrome: lethal chondrodysplasia associated with brain malformations in a 35-week-gestation infant. AB - This case report describes the neuropathological findings in an autopsy case of short rib-polydactyly syndrome (SRPS). The patient was a Japanese female neonate who was born at 35 weeks of gestation and died soon after birth due to severe cardiopulmonary insufficiency. Clinical and radiological findings were most consistent with SRPS type I (Saldino-Noonan type). General autopsy findings included situs inversus, persistent truncus arteriosus and endocardial cushion defect, hypoplastic lungs and adrenal glands, and vaginal atresia. Fixed brain weight was 330 g. Three different categories of pathological changes were detected in the brain. These were as follows: (1) multiple cyst formation in the parenchyma, (2) primary malformations of the nervous and mesenchymal tissues, and (3) deposition of an unusual substance in the cerebral white matter. The multiple cysts or cavities in the parenchyma may be due to severe hypoxic-ischemic insults related to the congenital heart anomaly. The primary malformations were summarized as follows: (1) capillary telangiectasia of the pia mater and choroid plexus, (2) olfactory dysplasia with asymmetry, (3) focal cortical dysplasia in the frontal lobe and cerebellum, (4) olivary dysplasia, and (5) enlargement of the posterior part of the lateral ventricle. Dysplastic changes of the nervous tissue can be classified into the group of neuronal migration disorders. Although biochemical properties of the unknown substance were not determined, it is considered to be some product derived from an inborn error of metabolism. Morphological data of SRPS is still scarce, and pathognomonic changes have not yet been elucidated. The present data suggests that coexistence of the nervous and mesenchymal malformations may be highly characteristic of SRPS. PMID- 16719408 TI - Fronto-laterally located supratentorial bronchogenic cyst: case report and review of the literature. AB - Bronchogenic cysts are rare findings within the central nervous system and even extremely seldom if they are located supratentorially. We report on a 25-year-old man who presented with a single generalized seizure without any accompanying neurological deficit 6 weeks prior to admission. MRI showed a smoothly limited, non-calcified cyst of a maximum of 55 mm in diameter. Large parts of the membrane were removed through a right anterior craniotomy, and a fenestration into the subarachnoidal space was performed. Histopathological examinations revealed a bronchogenic cyst. Previous reports of neurenteric cysts are reviewed. Therapeutic options, pathogenesis and categorization of this uncommon congenital entity are discussed. PMID- 16719410 TI - [Diagnosis and treatment of liver injuries]. AB - AIM: The aim is to specify contemporary diagnostic and therapeutic procedures of liver injuries. METHODOLOGY: The authors present a trial group of 117 patients with various stages of liver injuries hospitalized at the Traumacentrum Surgical Department of the Faculty Hospital in Plzen from 1.1.2000 to 31.9.2005. 61 injured (52.1%) were treated conservatively, 56 (47.9%) surgically, who underwent 87 procedures, in total. RESULTS: 10 polytrauma patients exited (8.5%) from a haemorrhagic shock (group Moore IV-VI). Six subjects (5.1%) had complications on hospitalization. Average hospitalization lasted 18 days (1-69 days). CONCLUSION: The basic diagnostic tools in liver injuries include "bed side" ultrasonography, spiral computer tomography, in more complicated traumas also MRI, angiography or endoscopic retrograde cholangiography. As a treatment strategy, conservative methods prevail in haemodynamically stable patients, regardless of the liver injury stage. The objective of the surgical management is to provide "damage control surgery", i.e. the first procedure is aimed at life-saving, no time demanding surgical procedures (e.g. liver tamponade), management of hypothermia, haemocoagulation and metabolic acidosis. PMID- 16719411 TI - [Prognostic factors of early recurrence of colorectal liver metastases and their usage in clinical praxis]. AB - Liver surgery for colorectal liver metastases (CLM) is very often followed by recurrence. The prediction of early relapse of CLM is important for successful treatment, long disease free period and for long survival of patients, because it could help us to choose adequate surgical and oncological treatment. The authors present contemporary overview of the prognostic factors of early CLM recurrence and their possible use in prognostic scoring systems. These could be very helpful in clinical practice. PMID- 16719412 TI - A prospective clinical feasibility study for multimodal "fast track" rehabilitation in elective pancreatic cancer surgery. AB - INTRODUCTION: Surgery on the pancreas is a major abdominal procedure leading to a number of pathophysiological alterations during the early post-operative period. Novel approaches to perioperative care including shortened pre-operative starving periods, pre-operative glucose load, sophisticated pain management and early enteral feeding have resulted in major improvements of surgical results after major colorectal surgery. These alterations of perioperative care have been introduced to visceral surgery as so-called fast track surgery or multimodal rehabilitation (multimodal rehabilitation, ERAS = enhanced recovery after surgery). So far it is not known whether or not these approaches can also be applied in pancreatic cancer surgery. METHODS: Twelve patients underwent fast track rehabilitation after pancreatic cancer surgery and their clinical course was compared with age-, sex-, and disease-matched control patients. In addition to clinical parameters (resumption of gastrointestinal function, complication rates, postoperative length of stay) we compared leukocyte counts and C-reactive protein serum levels of both patient groups. Patients recruited for this prospective study received clear carbohydrate rich fluid until two hours before surgery. Bowel preparation was reduced to one-time administration of a laxative and pain treatment consisted of thoracic epidural analgesia in combination with COX-II inhibitors. Intraoperative fluid administration was restricted to 500 cc of colloids and 500 cc of electrolytes. Oral food intake started on the day of surgery with clear fluids and was increased to a small amount of solid food on day 3 after surgery. Complete enteral nutrition was initiated on day 5 after surgery following opaque media examination of the upper gastrointestinal tract. Demission from hospital was planned on day 10 after surgery. RESULTS: The clinical course of patients undergoing fast track rehabilitation was significantly faster regarding resumption of bowel function and complete enteral nutrition. Furthermore, postoperative length of hospital stay was significantly shorter in fast track patients. CONCLUSION: While routine laboratory parameters showed similar changes during the postoperative course after pancreatic surgery, the clinical outcome parameters clearly indicated that the concept of fast track rehabilitation can be even beneficially applied to these high-risk patients undergoing elective surgery for pancreatic cancer. PMID- 16719413 TI - [The prepancreatic course of superior mesenteric vein--a case report]. AB - A case of a prepancreatic course of the superior mesenteric vein found at post mortem is described and a possible hypothesis of its development is offered. The prepancreatic course of the superior mesenteric or portal vein is not frequent, but in abdominal surgery it is necessary to take these anomalies into the consideration because of possible severe sequelae in the case of an inadvertent ligature of important vessels. PMID- 16719414 TI - [Re-laparotomy for complications of urgent abdominal disorders]. AB - Urgent abdominal disorders are a daily work routine at surgical departments. Therefore, all surgeons are familiar with the problematics, including all its pitfalls, e.g. high morbidity rates and mortality rates of the patients managed. The authors record urgent abdominal disorders mortality rates as well as rates of serious surgical complications requiring re-operations and their effect on the patients' prognosis. The trial group includes 1861 patients urgently operated during 2001-2004. 36 patients (1.9%) underwent re-laparotomies. Peritonitis in some form (50%), dehiscence of the laparotomy (31.2%), necrosis of the intestine, ileus, bleeding and some less frequent complications, were the most frequent indications for re-laparotomies. The thirty-day mortality rate in patients, operated for urgent abdominal disorders, was 4.8%. In the re-operated group, the mortality rate reached 19.4%. PMID- 16719415 TI - [Paliative managment of oesophageal carcinomas--our experience]. AB - The authors of this presentation explain their experience with methods of paliative care of oesophagus carcinoma. The indication criteria are described and compared advantages and disadvantages of those methods. There is an accent in most frequent method--oesophagus stent implementation. The results in the group of patients after the stent implementation are described, also complications and other solutions. PMID- 16719416 TI - Changing of the guard won't change the course of group purchasing. PMID- 16719417 TI - Suture prices stay the course in 2005. PMID- 16719418 TI - Cultures of safety that "stick": Sentara Norfolk and Self Regional. PMID- 16719419 TI - Communication isn't always about speaking English. PMID- 16719420 TI - Establishing a standard for pediatric inpatient diabetes education. AB - Education is a critical component of diabetes treatment and is crucial to achieve self-management outcomes in the pediatric patient. The education plan for new onset diabetes in a hospital setting must be goal oriented, consistent, and structured. Balancing the education needs, psychosocial response to diagnosis, family social support, and developmental needs of the child is often difficult considering hospital resource utilization and shortened length of stay (Leyden, Caravalho, & Saylor, 2000). Through analysis of current literature and current diabetes education processes within the hospital, a standard was developed that supports patient and family education from the moment of admission. The staff nurse is now the primary patient and family educator; therefore, educational opportunities are no longer compromised by off-shifts, weekends, and holidays, potentially increasing the length of stay. PMID- 16719421 TI - Insulin resistance syndrome in children. AB - Insulin resistance, a metabolic dysfunction in glucose metabolism, is associated with numerous physical health findings that have serious health consequences such as obesity, hyperlipidemia, hypertension, cardiovascular disease, and type 2 diabetes. The clustering of these diseases is termed insulin resistance syndrome, previously described as a metabolic condition exclusive to adults. Studies now indicate many of the components associated with insulin resistance syndrome are present in children and adolescents. Identifying children at risk for the development of insulin resistance is important for early prevention and intervention. Detecting early evidence of insulin resistance in children and adolescents can have a preventive effect on the disease components associated with insulin resistance. Primary care providers are in pivotal position for early recognition and treatment of insulin resistance in children. PMID- 16719422 TI - Management of pediatric patients with chronic kidney disease. AB - Chronic kidney disease in children is associated with complications that require nursing interventions in both the inpatient and outpatient settings. Given the progressive nature of the disease and the complexity of the treatment regimen, it is important that nurses be comfortable implementing acute and preventive care strategies and facilitating the coordination of care. In addition, the need for multiple therapies can be distressing for patients and their families, further supporting the role of the nurse in patient and family education and decision making regarding the plan of care. In this review, we discuss the pertinent issues of pediatric chronic kidney disease in the context of a case study to promote better understanding of real-world nursing practice. PMID- 16719424 TI - A well child visit. PMID- 16719423 TI - Caring for adoptive families: lessons in communication. AB - PURPOSE: To describe the experiences of families whose adopted children were hospitalized and to compare those experiences to the experiences of families of hospitalized biological children. METHOD: The parents of 10 adopted and 55 biological children participated in the study in which the source and intensity of stress for parents and the level of distress demonstrated by the children during their hospitalization were measured and compared. RESULTS: Adopted children hospitalized for the first time were significantly more distressed during hospitalization than biological children hospitalized for the first time. Adoptive parents experienced significantly more stress than biological parents related to how staff communicated with them about their child's illness. Concerns identified by several adoptive families included difficulties associated with having limited information about their child's family medical history and concerns about attachment issues. CONCLUSIONS: Health care providers need to be aware of adoptive parents' concerns about their child's response to hospitalization, attachment issues, and limited family medical history. The quality of communication with adoptive parents is especially important. PMID- 16719425 TI - Emergency contraception: a primer for pediatric providers. AB - Emergency contraception (EC) is a contraceptive method used safely and successfully by women for more than 30 years to prevent pregnancy. Nurses at all levels are often the first point of contact for a woman who is requesting EC, thus it is particularly important for them to stay abreast of both the facts regarding the use of this product and the current political controversies. It is particularly important for Nurse Practitioners (NPs) working in primary care with adolescents to remain cognizant of the significant barriers that remain for many women of all ages trying to access this important contraceptive tool. PMID- 16719426 TI - Evidence-based review of nursing interventions to prevent secondary disabilities in fetal alcohol spectrum disorder. PMID- 16719427 TI - Petting zoos. PMID- 16719428 TI - Pediatric health policy analysis: the Emergency Medical Services for Children (EMSC) Act and the Wakefield Act, utilizing social construction of target populations. PMID- 16719429 TI - In their own voices: families discuss end-of-life decision making--part 1. Interview by Elizabeth Ahmann. AB - One parent's experience with her daughter in a neonatal intensive care unit (NICU) and pediatric intensive care unit (PICU) setting sheds light on parental involvement in end-of-life decision making. For this parent, collaborative decision making was facilitated in an environment where the parents had gotten to know the health care providers over time. Health care provider style was a factor in providing parents with access to information needed to participate in decisions. Regular meetings away from the bedside facilitated parental information gathering and joint decision making. This parent also used outside advice in making key decisions. In contrast, interaction with a minimally communicative and authoritarian style health care professional added greatly to this parent's stress in an already difficult circumstance. These are her own words. PMID- 16719430 TI - Job satisfaction among neonatal nurses. AB - PURPOSE: The purpose of this study was to understand the job satisfaction of nurses who work in intensive care nurseries. The design used a convenience sample of eight nurses with an average of 11 years of active and current experience as neonatal intensive care nurses. METHOD/DATA COLLECTION: Human rights were protected according to the institutional guidelines. Data collection included semi-structured, intensive face-to-face interviews, observation, and field notes. The interviews were tape recorded and transcribed. Collaizzi's (1978) interpretation method was used to interpret and analyze the data using significant statements, formulated meanings, and clustering. Each participant was allowed to review the typed interview as one means of credibility. RESULTS: Analysis of the described experiences revealed that nurses were able to identify enough satisfying situations that compelled them to continue working in the neonatal intensive care unit (NICU). These rewards included compensation, team spirit, support from physicians, and advocacy. CONCLUSION: Knowledge of the factors that contribute to nurses' job satisfaction can provide a useful framework to implement policies to improve working conditions for nurses. PMID- 16719431 TI - The Initiative for Pediatric Palliative Care (IPPC) what is IPPC? IPPC is both an education and a quality. PMID- 16719432 TI - Specialty carve-outs address unique reinsurance issues. PMID- 16719433 TI - New tool estimates DM costs even without administrative claims data. PMID- 16719434 TI - Payer takes novel approach to Medicare risk with ancillary cap rates. PMID- 16719435 TI - Productivity still king in physician comp, but impact of resource utilization growing. PMID- 16719436 TI - Timely breast, cervical cancer screening improves with managed care penetration. PMID- 16719437 TI - Quadruplex-to-duplex transition of G-rich oligonucleotides probed by cationic water-soluble conjugated polyelectrolytes. AB - G-quartet DNA converts to duplex form in the presence of its complementary strand. This conformational change can be detected in real time by a homogeneous assay method based on the signal amplification of conjugated polyelectrolytes and the specific interaction of intercalating dyes with double-stranded DNA (dsDNA). The probe solution contains a cationic, conjugated polymer (CCP), G-quadruplex labeled with a fluorescein at the 5'-terminus (G-quadruplex-Fl), and ethidium bromide (EB). The addition of a complementary target results in the transition from G-quadruplex to duplex (dsDNA-Fl) and EB intercalation within the duplex structure. Excitation of the CCP leads to energy transfer from CCP to dsDNA-Fl (FRET-1) and then energy transfer from dsDNA-Fl to EB (FRET-2). Increasing the number of mismatched bases discourages dsDNA formation, which is detected in the assay. PMID- 16719438 TI - Catalytic reduction of NO to N2O by a designed heme copper center in myoglobin: implications for the role of metal ions. AB - The effects of metal ions on the reduction of nitric oxide (NO) with a designed heme copper center in myoglobin (F43H/L29H sperm whale Mb, CuBMb) were investigated under reducing anaerobic conditions using UV-vis and EPR spectroscopic techniques as well as GC/MS. In the presence of Cu(I), catalytic reduction of NO to N2O by CuBMb was observed with turnover number of 2 mol NO.mol CuBMb-1.min-1, close to 3 mol NO.mol enzyme-1.min-1 reported for the ba3 oxidases from T. thermophilus. Formation of a His-heme-NO species was detected by UV-vis and EPR spectroscopy. In comparison to the EPR spectra of ferrous-CuBMb-NO in the absence of metal ions, the EPR spectra of ferrous-CuBMb-NO in the presence of Cu(I) showed less-resolved hyperfine splitting from the proximal histidine, probably due to weakening of the proximal His-heme bond. In the presence of Zn(II), formation of a five-coordinate ferrous-CuBMb-NO species, resulting from cleavage of the proximal heme Fe-His bond, was shown by UV-vis and EPR spectroscopic studies. The reduction of NO to N2O was not observed in the presence of Zn(II). Control experiments using wild-type myoglobin indicated no reduction of NO in the presence of either Cu(I) or Zn(II). These results suggest that both the identity and the oxidation state of the metal ion in the CuB center are important for NO reduction. A redox-active metal ion is required to deliver electrons, and a higher oxidation state is preferred to weaken the heme iron proximal histidine toward a five-coordinate key intermediate in NO reduction. PMID- 16719439 TI - Assembly state of catalytic modules as chiral switches in asymmetric Strecker amino acid synthesis. AB - Self-assembled chiral polymetallic complexes often demonstrate novel properties as asymmetric catalysts. We report the three-dimensional structures of two such asymmetric catalysts (crystals A and B) for Strecker alpha,alpha-disubstituted amino acid synthesis. These complexes are constructed via assembly of the same chiral modules derived from d-glucose, but their assembly modes differ. The enantioselectivity in the Strecker reaction was dramatically switched, depending on which assembly mode was used: the catalyst generated in situ whose structure is represented by crystal B, or by crystal A. These findings provide insight into the functional importance of higher-order structures of an artificial catalyst. PMID- 16719440 TI - Rational design of a reversible pH-responsive switch for peptide self-assembly. AB - Peptide TZ1H, based on the heptad sequence of a coiled-coil trimer, undergoes fully reversible, pH-dependent self-assembly into long-aspect-ratio helical fibers. Substitution of isoleucine residues with histidine at the core d positions of alternate heptads introduces a mechanism by which self-assembly is coupled to the protonation state of the imidazole side chain. Circular dichroism spectroscopy, transmission electron microscopy, and microrheology techniques revealed that the self-assembly of TZ1H coincides with a distinct coil-helix conformational transition that occurs within a narrow pH range near the pKa of the imidazole side chains of the core histidine residues. PMID- 16719441 TI - Site-specific DNA photocleavage by rhodium intercalators analyzed by MALDI-TOF mass spectrometry. AB - The DNA photocleavage reaction of mismatch-selective Rh complexes has been analyzed by MALDI-TOF mass spectrometry as well as gel electrophoresis analysis of radioactively tagged oligonucleotides. Analogous results are obtained with these two techniques showing site-specific cleavage neighboring the mismatch to yield primarily 5'- and 3'-phosphate termini. Additional intermediates and products are observed, however, using MALDI-TOF analysis. MALDI-TOF mass spectrometry is seen to be particularly powerful in the analysis of DNA cleavage by site-specific molecules. The method requires no radioactive labeling, only little material, and analysis can be accomplished within minutes. Moreover, this mass spectral analysis of DNA cleavage yields direct information regarding products rather than simply the base pair site of cleavage. PMID- 16719442 TI - Direct writing of a conducting polymer with molecular-level control of physical dimensions and orientation. AB - Polymer nanostructures composed of poly(3-dodecylthiophene) (PDDT) have been directly written with control of polymer strand alignment and monolayer-by monolayer thickness down to a single molecular monolayer (2.6 nm). The molecularly ordered nanostructures were written on silicon oxide surfaces using thermal dip-pen nanolithography, where an atomic force microscope cantilever with integrated tip heater was precoated with solid PDDT. The PDDT was precisely deposited onto the surface when the tip temperature was set close to PDDT's melting temperature. PMID- 16719443 TI - Direct observation of a 1MLCT state by ultrafast transient absorption spectroscopy in Mo2(O2C-9-anthracene)4. AB - We have observed the excited-state dynamics of Mo2(O2C-9-anthracene)4 in THF using ultrafast transient UV/vis absorption spectroscopy. Following excitation at 514.5 nm into the metal-to-ligand charge-transfer (MLCT) state, transient absorption bands of the 1MLCT state appear at 410 and 610 nm. We assign these features to the 1MLCT state, which has a lifetime of approximately 10 ps. The decay of 1MLCT is accompanied by the formation of the triplet 3MLCT state, with an absorption band peaking at 410 nm. Excitation at 347 nm populates directly the ligand-centered (LC) state. From the rise of the 1MLCT band, the lifetime of the 1LC state is estimated to be less than 1 ps. PMID- 16719444 TI - Isolation and molecular structure of the organo-persulfuranes [12-S-6(C6)]. AB - Reactions of bis(2,2'-biphenylylene)sulfuranyl bis(tetrafluoroborate) [(8-S 4(C4)]2+ with organo-lithium reagents (PhLi and MeLi) gave bis(2,2' biphenylylene)di-C-substituted persulfuranes as a first persulfurane [12-S-6(C6)] having only carbon ligands. These compounds have been characterized by 1H and 13C NMR and mass spectroscopies. The structure of the dimethyl derivative was determined by X-ray crystallographic analysis, revealing that it has a distorted octahedral geometry with the two methyl ligands cis to each other, and subsequently, it was analyzed by an ab initio calculation. PMID- 16719445 TI - An efficiently extended class of unnatural base pairs. AB - A third DNA base pair, which is synthesized efficiently and selectively, would have wide ranging applications from synthetic organisms to nucleic acids biotechnology. Hydrophobic unnatural nucleobases offer a promising route to such a pair, but are often limited by inefficient extension, defined as synthesis immediately following the unnatural pair. Here, we describe a simple screen which enables the characterization of large numbers of previously uncharacterized hetero base pairs. From this screen, we identified a class of unnatural base pairs which are extended more efficiently than any unnatural base pair reported to date. Screening, when complemented by further kinetic analysis, can improve the understanding of the determinants of efficient extension as well as identify viable hetero base pairs. PMID- 16719446 TI - Nuclear Overhauser effect (NOE) enhancement of 11B NMR spectra of borane adducts in the solid state. AB - A strong 11B {1H} nuclear Overhauser effect (NOE) enhancement can be observed in solid-state 11B NMR spectra of borane adducts, yielding fractional enhancements, fI{S} = (I - I0)/I0, of the magic angle spinning (MAS) NMR signal of up to 155%. This is an interesting and unusual observation as 11B (spin I = 3/2) is a quadrupolar nucleus and the corresponding NOE is completely absent in solution. More generally, it shows that the NOE may have a wider role to play in solid state NMR studies of dynamics than has been envisaged hitherto. PMID- 16719447 TI - A highly selective and sensitive inorganic/organic hybrid polymer fluorescence "turn-on" chemosensory system for iron cations. AB - A polymer-based fluorescence "turn-on" chemosensor is demonstrated with high sensitivity and selectivity toward Fe2+ cations. This system is an inorganic/organic hybrid system in which Cu2+ is coordinated at the receptor site of a conjugated polymer sensor. The unique quenching behavior of the Cu2+ toward this conjugated polymer was used to lower the initial fluorescence intensity of the sensory system. This results in a highly sensitive and also selective sensory system toward Fe2+ as compared to other related divalent transition-metal cations in solution. PMID- 16719448 TI - Pt-catalyzed tandem epoxide fragmentation/pentannulation of propargylic esters. AB - A Pt-catalyzed pentannulation of propargylic esters containing an epoxide moiety has been developed. The present transformation achieves the formation of cyclopentenone products as single diastereomers in good yields. The observed products likely form from pyran intermediates that undergo an oxa-6pi electrocyclic ring opening to a functionalized dienone, followed by ring closure with an accompanying acyl shift. PMID- 16719449 TI - Significant enhancement of monooxygenase activity of oxygen carrier protein hemocyanin by urea. AB - Oxygenation of a series of p-substituted phenols to the corresponding catechols (phenolase activity) by the (mu-eta2:eta2-peroxo)dicopper(II) species of Octopus hemocyanin has been directly examined for the first time by using a UV-vis spectroscopic method in a 0.5 M borate buffer solution containing 8 M urea under anaerobic conditions. Preliminary kinetic studies have indicated that the reaction involves an electrophilic aromatic substitution mechanism as in the case of phenolase reaction of tyrosinase. The oxygenation of phenols by hemocyanin also proceeded catalytically when the reaction was carried out under aerobic conditions. PMID- 16719450 TI - Cu(II)-catalyzed functionalizations of aryl C-H bonds using O2 as an oxidant. AB - Cu(II)-catalyzed acetoxylation and halogenation of aryl C-H bonds are developed. ortho-Selectivity was observed with a wide range of 2-arylpyridine substrates. Both mono- and difunctionalizations are achieved by tuning the reaction conditions. Excellent functional group tolerance and use of O2 as a stoichiometric oxidant are significant advantages over our recently developed Pd catalyzed C-H functionalization reactions. These newly discovered reaction conditions are also applicable for cyanation, amination, etherification, and thioetherification of aryl C-H bonds. Mechanistic investigations are carried out to gain insights into the Cu(II)-catalyzed C-H functionalization reactions. PMID- 16719451 TI - The hidden role of acetate in the PbSe nanocrystal synthesis. AB - Monodisperse spherical, star-shaped, and octahedral PbSe nanocrystals were synthesized via a hot injection method. We show that the shape and size of the colloidal PbSe nanocrystals are determined by the concentration of acetate and that only acetate-free reaction mixtures result in spherical nanocrystals. The presence of acetate leads to efficient oriented attachment of smaller PbSe nanoparticles along the 100 crystal axis. Comparing different synthesis procedures from the recent literature with our observations, we propose that the acetate, naturally present in insufficiently dried reaction mixtures, is responsible for many of the PbSe crystal shapes reported in the literature. In addition we show that it is possible to synthesize these star-shaped nanocrystals so monodisperse that they form ordered monolayers with crystal alignment. PMID- 16719452 TI - Three-dimensional DNA crystals as molecular sieves. AB - DNA has proved to be a successful nanoscale building block because of its inherent programmability and its predictable structural features. One long standing goal of DNA nanotechnology has been the rational design and assembly of three-dimensional (3D) DNA crystals for use as molecular scaffolds, in molecular electronics assembly, and as molecular sieves. Here we demonstrate that rationally designed 3D DNA crystals with mesoporous features can function as molecular sieves by selectively adsorbing proteins based on size. PMID- 16719453 TI - Triphasic nanocolloids. AB - Triphasic nanocolloids, that is, nanocolloids with three distinct compartments, were successfully produced by use of electrified co-jetting. Simultaneous manipulation of the three parallel liquid with laminar flows yielded a liquid droplet with three interfaces between the jetting liquids. Under a high electric potential, a single liquid jet was produced from the triple point of the droplet. Separation between the phases was maintained throughout the jetting process and resulted in triphasic nanocolloids. Poly(ethylene oxide), poly(acrylic acid), and poly(acrylamide-co-acrylic acid) solutions were used and verified to be compatible with the process. Fluorescent-labeled biomolecules were selectively incorporated in each phase of the nanocolloids, and confocal laser scanning microscopy was employed for the characterization. Scanning electron microscopy results verified that the sizes of the colloids are in the range of nanocolloids. This simple and versatile technique to fabricate multicompartment nanocolloids is expected to have great impact on drug delivery, molecular imaging, and smart displays. PMID- 16719455 TI - all-anti-octasilane: conformation control of silicon chains using the bicyclic trisilane as a building block. AB - The perfect all-anti-octasilane composed of two bicyclic trisilane units with trimethylsilyl groups at the termini has been synthesized. Its X-ray crystal structure and spectroscopic data demonstrate the effective sigma-delocalization over the silicon framework, which is a definitive difference from the unconstrained n-Si8Me18. PMID- 16719454 TI - Room temperature ring-opening metathesis of pyridines by a transient Ti[triple bond]C linkage. AB - The transient titanium alkylidyne complex (PNP)TiCtBu (PNP = N[2-P(CHMe2)2-4 methylphenyl]2-), prepared from alpha-hydrogen abstraction of the corresponding alkylidene-alkyl species (PNP)Ti=CHtBu(CH2tBu), can readily cleave the C-N bond of N-heterocycles such as pyridine and 4-picoline at room temperature to afford azametallabicyclic systems. Experimental and theoretical studies strongly favor a ring-opening metathesis pathway where [2 + 2] cycloaddition of pyridine across the TiC linkage ultimately leads to C-N bond rupture. PMID- 16719456 TI - Cyclocarbophosphazene-containing tetrameric assemblies formed by the mediation of P-O-P and P-O-Cu linkages. AB - Cyclocarbophosphazene-containing tetrameric assemblies formed by the mediation of P-O-P and P-O-Cu linkages have been isolated and characterized. PMID- 16719458 TI - Construction of double-stranded metallosupramolecular polymers with a controlled helicity by combination of salt bridges and metal coordination. AB - We describe the construction of the first double-stranded metallosupramolecular helical polymers. We designed and synthesized a supramolecular duplex comprised of complementary m-terphenyl-based strands bearing a chiral amidine or achiral carboxylic acid together with two pyridine groups at the four ends. Supramolecular polymerization of the duplex with cis-PtPh2(DMSO)2 in 1,1,2,2 tetrachloroethane produced the double-stranded metallosupramolecular polymer with a controlled helicity of which the two complementary metallostrands are intertwined through the amidinium-carboxylate salt bridges. The structures and hydrodynamic dimensions of the metallosupramolecular polymers were characterized by 1H NMR, diffusion-ordered NMR, dynamic light scattering, absorption, and CD measurements. The polymeric structure was also visualized by atomic force microscopy. PMID- 16719457 TI - Enantioselective organocatalytic aminomethylation of aldehydes: a role for ionic interactions and efficient access to beta2-amino acids. AB - Organocatalytic Mannich addition of aldehydes to a formaldehyde-derived iminium species catalyzed by proline-derived chiral pyrrolidines provides beta-amino aldehydes with >/=90% ee. Mechanistic analysis of the proline-catalyzed reactions suggests that non-hydrogen-bonded ionic interactions at the Mannich reaction transition state can influence stereochemical outcome. The beta-amino aldehydes from our process bear a substituent adjacent to the carbonyl and can be efficiently converted to protected beta2-amino acids, which are important building blocks for beta-peptide foldamers that display useful biological activities. PMID- 16719459 TI - Tandem synthesis of core-shell brush copolymers and their transformation to peripherally cross-linked and hollowed nanostructures. AB - Core-shell brush copolymers were prepared on the basis of a tandem synthetic strategy and used as single molecular templates for the preparation of polymeric nanomaterials. An alkoxyamine-functionalized norbornene monomer was prepared and then polymerized by ring-opening metathesis polymerization. The well-defined polymer (Mn = 122 kDa, Mw/Mn = 1.13) contained one alkoxyamine functionality per repeat unit and was then used as a polyfunctional macroinitiator for sequential nitroxide-mediated radical polymerizations of isoprene and tert-butyl acrylate. The resulting well-defined brush copolymer (Mn = 1410 kDa, Mw/Mn = 1.23) was transformed to an amphiphilic core-shell brush copolymer comprising poly(isoprene)-b-poly(acrylic acid) grafts by hydrolysis. Subsequent cross linking of the poly(acrylic acid) block segments afforded peripherally cross linked brush copolymer nanostructures, which served, finally, as templates for hollowed nanoscale frameworks by ozonolysis of the poly(isoprene)-based cores. Each transformation led to dramatic changes in the nanoscale composition and structure which were detected by combinations of spectroscopic measurements, atomic force microscopy imaging in the solid state, and/or dynamic light scattering characterization in aqueous solution. PMID- 16719460 TI - Alpha,beta-unsaturated beta-silyl imide substrates for catalytic, enantioselective conjugate additions: a total synthesis of (+)-lactacystin and the discovery of a new proteasome inhibitor. AB - Chiral (salen)Al mu-oxo dimer 1 catalyzes the highly enantioselective conjugate addition of carbon-centered nucleophiles to alpha,beta-unsaturated silyl imides. Allyldimethylsilane-substituted imide 4 was identified as an optimal substrate, undergoing addition reactions with a variety of nitrile nucleophiles in high yield and enantiomeric excess. The silicon-containing products are synthetically useful chiral building blocks, as demonstrated by their application to an enantioselective total synthesis of the potent proteasome inhibitor (+) lactacystin (2). Elaboration of lactam 5a to the natural product was effected in 12 steps and in 11% overall yield and proceeded through an unusual spiro beta lactone intermediate (11). This compound was found to inhibit the chymotrypsin like site of the 26S proteasome at similar levels to known inhibitor clasto lactacystin beta-lactone (omuralide). PMID- 16719461 TI - Determination of the orientation of adsorbed cytochrome C on carboxyalkanethiol self-assembled monolayers by in situ differential modification. AB - The contact domain utilized by horse cytochrome c when adsorptively bound to a C(10)COOH self-assembled monolayer (SAM) was delineated using a chemical method based on differential modification of surface amino acids. Horse cytochrome c was adsorbed at low ionic strength (pH 7.0, 4.4 mM potassium phosphate) onto 10 microm diameter gold particles coated with HS(CH(2))(10)COOH SAMs. After in situ modification of lysyl groups by reductive Schiff-base methylation, the protein was desorbed, digested using trypsin, and the peptide mapped using LC/MS. Relative lysyl reactivities were ascertained by comparing the resulting peptide frequencies to control samples of solution cytochrome c modified to the same average extent. The least reactive lysines in adsorbed cytochrome c were found to be 13, 72, 73, 79, and 86-88, consistent with a contact region located up and to the left (Met-80 side) of the solvent-exposed heme edge (conventional front face view). The most reactive lysines were 39, 53, 55, and 60, located on the lower backside. The proposed orientation features a heme tilt angle of approximately 35 40 degrees with respect to the substrate surface normal. Factors that can complicate or distort data interpretation are discussed, and the generality of differential modification relative to existing in situ methods for protein orientation determination is also addressed. PMID- 16719462 TI - Indirect NMR spin-spin coupling constants 3J(P,C) and 2J(P,H) across the P-O...H C link can be used for structure determination of nucleic acids. AB - Calculated indirect NMR spin-spin coupling constants (3)J(P,C) and (2)J(P,H) were correlated with the local structure of the P-O...H-C linkage between the nucleic acid (NA) backbone phosphate and the H-C group(s) of a nucleic acid base. The calculations were carried out for selected nucleotides from the large ribosomal subunit (Ban et al. Science 2000, 289, 905) with the aim of identifying NMR parameters suitable for detection of certain noncanonical RNA structures. As calculations in the model system, dimethyl-phosphate-guanine, suggest, the calculated indirect spin-spin couplings across the linkage are sensitive to the mutual orientation and distance between the phosphate and nucleic acid base. A short distance between the nucleic acid base and phosphate group and the angles C...P-O and P...C-H smaller than 50 degrees are prerequisites for a measurable spin-spin interaction of either coupling (|J| > 1 Hz). A less favorable arrangement of the P-O...H-C motif, e.g., in nucleotides of the canonical A-RNA, results in an effective dumping of both spin-spin interactions and insignificant values of the NMR coupling constants. The present work indicates that quantum chemical calculations of the indirect spin-spin couplings across the P-O...H-C motif can help detect some rare but important backbone topologies, as seen for example in the reverse kink-turn. Measuring of (3)J(P,C) and (2)J(P,H) couplings can therefore provide critical constraints on the NA base and phosphate geometry and help to determine the structure of NAs. PMID- 16719463 TI - Mechanism of the palladium-catalyzed homocoupling of arylboronic acids: key involvement of a palladium peroxo complex. AB - The mechanism of the palladium-catalyzed homocoupling of arylboronic acids ArB(OH)(2) (Ar = 4-Z-C(6)H(4) with Z = MeO, H, CN) in the presence of dioxygen, leading to symmetrical biaryls, has been fully elucidated. The peroxo complex (eta(2)-O(2))PdL(2) (L = PPh(3)), generated in the reaction of dioxygen with the Pd(0) catalyst, was found to play a crucial role. Indeed, it reacts with the arylboronic acid to generate an adduct (coordination of one oxygen atom of the peroxo complex to the oxophilic boron atom of the arylboronic acid) characterized by (31)P NMR spectroscopy and ab initio calculations. This adduct reacts with a second molecule of arylboronic acid to generate trans-ArPd(OH)L(2) complexes. A transmetalation by the arylboronic acid gives trans-ArPdArL(2) complexes. The biaryl is then released in a reductive elimination. This reaction is at the origin of the formation of biaryls as byproducts in palladium-catalyzed Suzuki Miyaura reactions when they are not conducted under oxygen-free atmosphere. PMID- 16719464 TI - Rhodium-catalyzed ring-opening reactions of N-boc-azabenzonorbornadienes with amine nucleophiles. AB - In the presence of a rhodium catalyst (5 mol %) generated in situ from [Rh(cod)Cl](2) and (S,S')-(R,R')-C(2)-ferriphos (4a), the asymmetric ring-opening reaction of azabenzonorbornadienes (1a-m) with various aliphatic and aromatic amines (2a-l) proceeded with high enantioselectivity (up to >99% ee) to give the corresponding 1,2-diamine derivatives 3 in high yields. In the specific case of pyrrolidine as nucleophile, Et(3)NHCl was necessary as an additive for good reactivity and enantioselectivity. Additionally, a practical protocol was developed for the ring-opening of 1a with volatile amines at elevated temperatures and standard pressure, using R(2)NH(2)I and i-Pr(2)NEt. The experimental results showed that the nature of the chiral ligand has the significant impact on the reactivity of the catalyst and the use of excess amount (2.2 eq to Rh) of the chiral ligand plays an important role to improve the enantioselectivity in the present asymmetric reaction. PMID- 16719465 TI - Antiparallel-aligned neutral-ground-state and zwitterionic chromophores as a nonlinear optical material. AB - Efficient noncentrosymmetric arrangement of nonlinear optical (NLO) chromophores with high first-order hyperpolarizability (beta) for increased electro-optical (EO) efficiency has proven challenging as strong dipolar interactions between the chromophores encourage antiparallel alignment, attenuating the macroscopic EO effect. This work explores a novel approach to simultaneously achieve large beta values while providing an adjustable dipole moment by linking a strong neutral ground-state (NGS) NLO chromophore with positive beta to a zwitterionic (ZWI) chromophore with negative beta in an antiparallel fashion. It is proposed that the overall beta of such a structure will be the sum of the absolute values of the two types of chromophores while the dipole moment will be the difference. Molecules 1-3 were synthesized to test the feasibility of this approach. Molecular dynamics calculations and NMR data supported that the NGS chromophore component and the ZWI chromophore component self-assemble to an antiparallel conformation in chloroform. Calculations showed that the dipole moment of 1 is close to the difference of the two component chromophores. Hyper-Rayleigh scattering (HRS) studies confirmed that the first hyperpolarizability of 1 is close to the sum of the two component chromophores. These results support the idea that an antiparallel-aligned neutral-ground-state chromophore and a zwitterionic chromophore can simultaneously achieve an increase in beta and a decrease of the dipole moment. PMID- 16719466 TI - Controlling semiconductor/metal junction barriers by incomplete, nonideal molecular monolayers. AB - We study how partial monolayers of molecular dipoles at semiconductor/metal interfaces can affect electrical transport across these interfaces, using a series of molecules with systematically varying dipole moment, adsorbed on n GaAs, prior to Au or Pd metal contact deposition, by indirect evaporation or as "ready-made" pads. From analyses of the molecularly modified surfaces, we find that molecular coverage is poorer on low- than on high-doped n-GaAs. Electrical charge transport across the resulting interfaces was studied by current-voltage temperature, internal photoemission, and capacitance-voltage measurements. The data were analyzed and compared with numerical simulations of interfaces that present inhomogeneous barriers for electron transport across them. For high-doped GaAs, we confirm that only the former, molecular dipole-dependent barrier is found. Although no clear molecular effects appear to exist with low-doped n-GaAs, those data are well explained by two coexisting barriers for electron transport, one with clear systematic dependence on molecular dipole (molecule-controlled regions) and a constant one (molecule-free regions, pinholes). This explains why directly observable molecular control over the barrier height is found with high doped GaAs: there, the monolayer pinholes are small enough for their electronic effect not to be felt (they are "pinched off"). We conclude that molecules can control and tailor electronic devices need not form high-quality monolayers, bind chemically to both electrodes, or form multilayers to achieve complete surface coverage. Furthermore, the problem of stability during electron transport is significantly alleviated with molecular control via partial molecule coverage, as most current flows now between, rather than via, the molecules. PMID- 16719467 TI - Synthesis of fluoranthenes and indenocorannulenes: elucidation of chiral stereoisomers on the basis of static molecular bowls. AB - Cycloaromatization of a peri diyne with an external alkyne provides a general route to indeno-fused polynuclear aromatics. Fluoranthenes 9 (or 13) are easily accessible in good to excellent yields (75-99%; 18 examples) from the reaction of symmetric (or asymmetric) diynes 4 and alkynes 5 (or norbornadiene) in the presence of Wilkinson's catalyst. This formal [(2+2)+2] cycloaddition can also be applied to generate various indenocorannulenes 28 from 2,3-diethynylcorannulene derivatives 27 and alkynes 5. The indenocorannulenes 28 exist in a static bowl form at room temperature with bowl-to-bowl inversion barriers higher than 24 kcal/mol. This barrier renders the rate of inversion slow enough at room temperature to establish a class of chiral, bowl-shaped stereoiomers containing no tetrahedrally ligated atoms. The crystal structure of 28g provides insight into the bowl-shaped geometry of these compounds. This new synthetic method occurs under neutral conditions and tolerates various functional groups (e.g., alkyl, aryl, alcohols, and esters). PMID- 16719468 TI - Ultrafast dynamics in DNA: "fraying" at the end of the helix. AB - The dynamics of the electric fields in the interior of DNA are measured by using oligonucleotides in which a native base pair is replaced by a dye molecule (coumarin 102) whose emission spectrum is sensitive to the local electric field. Time-resolved measurements of the emission spectrum have been extended to a six decade time range (40 fs to 40 ns) by combining results from time-correlated photon counting, fluorescence up-conversion, and transient absorption. Recent results showed that when the reporter is placed in the center of the oligonucleotide, the dynamics are very broadly distributed over this entire time range and do not show specific time constants associated with individual processes (Andreatta, D.; et al. J. Am. Chem. Soc. 2005, 127, 7270). This paper examines an oligonucleotide with the reporter near its end. The broadly distributed relaxation seen before remains with little attenuation. In addition, a new relaxation with a well-defined relaxation time of 5 ps appears. This process is assigned to the rapid component of "fraying" at the end of the helix. PMID- 16719469 TI - Flexible biphenyl chromophore as a circular dichroism probe for assignment of the absolute configuration of carboxylic acids. AB - A general and nonempirical approach to determine the absolute configuration (AC) of 2-substituted chiral carboxylic acids by circular dichroism (CD) spectroscopy has been developed. In this protocol, the chiral acids are converted to the corresponding biphenyl amides, in which a flexible biphenyl probe gives rise to a Cotton effect at 250 nm (A band) in the CD spectrum, the sign of which is related to the acid AC. Two different mechanisms of transfer of chirality from the acid stereogenic center to the biphenyl moiety are operative in amides derived from 2 alkyl- and 2-aryl-substituted acids, respectively. For both classes of compounds, a model has been defined which allows one to predict, for a given acid AC, the preferred twist of the biphenyl moiety and thus the sign of the A band in the CD spectrum, related to the biphenyl torsion. Interestingly, while in alkyl substituted substrates the preferred biphenyl twist is determined only by steric interactions, in the aryl-substituted ones the structure of the prevalent conformer and thus the biphenyl twist are dictated by arene-arene edge-to-face stabilizing interactions. Following this protocol, the AC of a 2-substituted chiral acid can be established simply by preparing its biphenyl amides, recording the CD spectrum, and looking at the sign of the A band. From the sign of such a band, the torsion of the biphenyl can be deduced and then the acid AC. Substrates having different structures and functionalities have been investigated, always obtaining reliable AC assignments by this simple protocol. PMID- 16719470 TI - Direct observation of the release of phenylalanine from diphenylalanine nanotubes. AB - The core recognition motif of the amyloidogenic beta-amyloid polypeptide is a dipeptide of phenylalanine. This dipeptide readily self-assembles to form discrete, hollow nanotubes with high persistence lengths. The simplicity of the nanotube formation, combined with ideal physical properties, make these nanotubes highly desirable for a range of applications in bionanotechnology. To fully realize the potential of such structures, it is first necessary to gain a comprehensive understanding of their chemical and physical properties. Previously, the thermal stability of these nanotubes has been investigated by electron microscopy. Here, we further our understanding of the structural stability of the nanotubes upon dry-heating using the atomic force microscope (AFM), and for the first time identify their degradation product utilizing time of-flight secondary-ion mass spectrometry. We show that the nanotubes are stable at temperatures up to 100 degrees C, but on heating to higher temperatures begin to lose their structural integrity with an apparent collapse in tubular structure. With further increases in temperature up to and above 150 degrees C, there is a degradation of the structure of the nanotubes through the release of phenylalanine building blocks. The breakdown of structure is observed in samples that are either imaged at elevated temperatures or imaged following cooling, suggesting that once phenylalanine is lost from the nanotubes they are susceptible to mechanical deformation by the imaging AFM probe. This temperature induced plasticity may provide novel properties for these peptide nanotubes, including possible applications as scaffolds and drug delivery devices. PMID- 16719471 TI - Solid-state NMR study of the transformation of octacalcium phosphate to hydroxyapatite: a mechanistic model for central dark line formation. AB - For many years, octacalcium phosphate (OCP) has been postulated as the precursor phase of biological apatite in bones and teeth. In this work, we study the molecular mechanism of OCP to hydroxyapatite (HAp) transformation in vitro by several physical techniques, with particular emphasis on solid-state (31)P homonuclear double-quantum (DQ) NMR spectroscopy. The in vitro system is prepared by mixing urea, sodium phosphate monobasic dehydrate, and calcium nitrate tetrahydrate at 100 degrees C. The images obtained by scanning electron microscopy and transmission electron microscopy show that the bladelike OCP crystals will transform into hexagonal rod-shaped HAp crystals as the pH of the reaction mixture increases slowly from 4.35 to 6.69 in 12 h. Powder X-ray diffraction patterns indicate that a trace amount of monetite was also precipitated when the pH was around 5. Together with computer-assisted lattice matching, our DQ NMR data reveal that OCP crystals transform to HAp topotaxially, where the [000](HAp) and [20](HAp) axes are along the same directions as the [001](OCP) and [010](OCP) axes, respectively. On the basis of our in vitro results, the formation of the central dark line commonly found in biological hard tissues could be explained by the inherent lattice mismatch between OCP and HAp. Furthermore, the data of the (31)P{(1)H} cross-polarization NMR suggest that water molecules enter the hydration layers of OCP crystals via the hydrolysis reaction HPO(4)(2)(-) + OH(-) = PO(4)(3)(-) + H(2)O, which also accounts for the deprotonation of the HPO(4)(2)(-) ions during the transformation. PMID- 16719472 TI - Synthesis of butane-like SiGe hydrides: enabling precursors for CVD of Ge-rich semiconductors. AB - The synthesis of butane-like (GeH(3))(2)(SiH(2))(2) (1), (GeH(3))(2)SiH(SiH(3)) (2), and (GeH(3))(2)(SiH(2)GeH(2)) (3) Si-Ge hydrides with applications in low temperature synthesis of Ge-rich Si(1-x)Ge(x) optoelectronic alloys has been demonstrated. The compositional, vibrational, structural, and thermochemical properties of these compounds were studied by FTIR, multinuclear NMR, mass spectrometry, Rutherford backscattering, and density functional theory (DFT) simulations. The analyses indicate that the linear (GeH(3))(2)(SiH(2))(2) (1) and (GeH(3))(2)(SiH(2)GeH(2)) (3) compounds exist as a mixture of the classic normal (n) and gauche (g) conformational isomers which do not seem to interconvert at 22 degrees C. The conformational proportions in the samples were determined using a new fitting procedure, which combines calculated molecular spectra to reproduce those observed by varying the global intensity, frequency scale, and admixture coefficients of the individual conformers. The (GeH(3))(2)(SiH(2))(2) (1) species was then utilized to fabricate Si(0.50)Ge(0.50) semiconductor alloys reflecting exactly the Si/Ge content of the precursor. Device quality layers were grown via gas source MBE directly on Si(100) at unprecedented low temperatures 350-450 degrees C and display homogeneous compositional and strain profiles, low threading dislocation densities, and atomically planar surfaces. Low energy electron microscopy (LEEM) analysis has demonstrated that the precursor is highly reactive on Si(100) surfaces, with H(2) desorption kinetics comparable to those of Ge(2)H(6), despite the presence of strong Si-H bonds in the molecular structure. PMID- 16719473 TI - Synthesis of (-)-sordarin. AB - The first total synthesis of (-)-sordarin (1) was accomplished exploiting the following key reactions: (i) Ag(I)-catalyzed oxidative radical cyclization of a cyclopropanol derivative leading to a bicyclo[5.3.0]decan-3-one skeleton; (ii) Pd(0)-catalyzed intramolecular allylation reaction resulting in the entire strained bicyclo[2.2.1]heptan-2-one framework of sordaricin (2); (iii) selective dihydroxylation of terminal alkenes by the combined use of OsO(4) and PhB(OH)(2); and (iv) beta(1,2-cis)-selective glycosidation via a 1,3-anchimeric assistance from a 4-methoxybenzoyl group. PMID- 16719474 TI - Modulation of luminescence intensity of lanthanide complexes by photoinduced electron transfer and its application to a long-lived protease probe. AB - Luminescent lanthanide complexes (Tb(3+), Eu(3+), etc.) have excellent properties for biological applications, including extraordinarily long lifetimes and large Stokes shifts. However, there have been few reports of lanthanide-based functional probes, because of the difficulty in designing suitable complexes with a luminescent on/off switch. Here, we have synthesized a series of complexes which consist of three moieties: a lanthanide chelate, an antenna, and a luminescence off/on switch. The antenna is an aromatic ring which absorbs light and transmits its energy to the metal, and the switch is a benzene derivative with a different HOMO level. If the HOMO level is higher than a certain threshold, the complex emits no luminescence at all, which indicates that the lanthanide luminescence can be modulated by photoinduced electron transfer (PeT) from the switch to the sensitizer. This approach to control lanthanide luminescence makes possible the rational design of functional lanthanide complexes, in which the luminescence property is altered by a biological reaction. To exemplify the utility of our approach to the design of lanthanide complexes with a switch, we have developed a novel protease probe, which undergoes a significant change in luminescence intensity upon enzymatic cleavage of the substrate peptide. This probe, combined with time-resolved measurements, was confirmed in model experiments to be useful for the screening of inhibitors, as well as for clinical diagnosis. PMID- 16719476 TI - Quantitative structure-activity relationships of ruthenium catalysts for olefin metathesis. AB - A quantitative structure-activity relationship (QSAR) model is presented in which both the independent and dependent (response) variables are derived from density functional theory (DFT) calculations on a large set of 14-electron complexes, LCl(2)Ru=CH(2), with different dative ligands, L. The multivariate model thus correlates the properties of the 14-electron complexes with a calculated measure of activity, with modest computational cost, and reproduces the experimental order of activity for the Grubbs ruthenium catalysts for olefin metathesis. The accuracy and applicability of the model is to a large extent due to the use of highly specific geometric and electronic molecular descriptors which establish a direct connection between activity and chemically meaningful donor ligand properties. The ligands that most efficiently promote catalytic activity are those that stabilize the high-oxidation state (+4) metallacyclobutane intermediate relative to the ruthenium-carbene structures dominating the rest of the reaction pathway. Stabilization of the intermediate is ensured, among others, through ligand-to-metal sigma donation, whereas metal-to-ligand pi back-donation destabilizes the intermediate and lowers catalytic activity. A bulky dative ligand drives the reaction toward the less sterically congested metallacyclobutane species and thus contributes to catalytic activity. The multivariate model and the high-level descriptors furthermore provide practical handles for catalyst development as exemplified by the suggestion of several new donor ligands predicted to give more active and functional group tolerant ruthenium-based catalysts. The present strategy holds great promise for broader screenings of olefin metathesis catalysts as well as for development of homogeneous transition metal catalysts in general. PMID- 16719477 TI - Stepwise formation and characterization of covalently linked multiporphyrin-imide architectures on Si(100). AB - A major challenge in molecular electronics and related fields entails the fabrication of elaborate molecular architectures on electroactive surfaces to yield hybrid molecular/semiconductor systems. A method has been developed for the stepwise synthesis of oligomers of porphyrins linked covalently via imide units. A triallyl-porphyrin bearing an amino group serves as the base unit on Si(100), and the alternating use of a dianhydride (3,3',4,4'-biphenyltetracarboxylic dianhydride) and a porphyrin-diamine for reaction enables the rapid and simple buildup of oligomers composed of 2-5 porphyrins. The properties of these porphyrin "multad" films on Si(100) were interrogated using a variety of techniques. The charge densities of the redox-active porphyrin oligomers were determined via electrochemical methods. The stepwise growth was evaluated in detail via Fourier transform infrared (FTIR) spectroscopy and by selected X-ray photoelectron spectroscopic (XPS) studies. The morphology was probed via AFM methods. Finally, the thickness was evaluated by using a combination of ellipsometry and AFM height profiling, accompanied by selected XPS studies. Collectively, these studies demonstrate that high charge density, ultrathin, multiporphyrin films of relatively well-controlled thickness can be grown in a stepwise fashion using the imide-forming reaction. The increased charge densities afforded by the porphyrin multads may prove important for the fabrication of molecular-based information-storage devices. This bottom-up process for construction of surface-tethered molecular architectures complements the top-down lithographic approach for construction of functional devices with nanoscale dimensions. PMID- 16719475 TI - Increasing the accuracy of solution NMR structures of membrane proteins by application of residual dipolar couplings. High-resolution structure of outer membrane protein A. AB - The structure determination of membrane proteins is one of the most challenging applications of solution NMR spectroscopy. The paucity of distance information available from the highly deuterated proteins employed requires new approaches in structure determination. Here we demonstrate that significant improvement in the structure accuracy of the membrane protein OmpA can be achieved by refinement with residual dipolar couplings (RDCs). The application of charged polyacrylamide gels allowed us to obtain two alignments and accurately measure numerous heteronuclear dipolar couplings. Furthermore, we have demonstrated that using a large set of RDCs in the refinement can yield a structure with 1 A rms deviation to the backbone of the high-resolution crystal structure. Our simulations with various data sets indicate that dipolar couplings will be critical for obtaining accurate structures of membrane proteins. PMID- 16719478 TI - The properties of the [Mn12O12(O2CR)16(H2O)4] single-molecule magnets in truly axial symmetry: [Mn12O12(O2CCH2Br)16(H2O)4].4CH2Cl2. AB - Detailed studies are reported of a Mn(12) single-molecule magnet (SMM) in truly axial (tetragonal) symmetry. The complex is [Mn(12)O(12)(O(2)CCH(2)Br)(16)(H(2)O)(4)].4CH(2)Cl(2) (2.4CH(2)Cl(2) or Mn(12) BrAc), obtained by the standard carboxylate substitution method. The complex has an S = 10 ground state, typical of the Mn(12) family, and displays frequency dependent out-of-phase AC susceptibility signals and hysteresis in single-crystal magnetization vs applied DC field sweeps. Single-crystal high-frequency EPR spectra in frequencies up to 360 GHz exhibit narrow signals that are not overlapping multiplets, in contrast to [Mn(12)O(12)(O(2)CMe)(16)(H(2)O)(4)].2MeCO(2)H.4H(2)O (1 or Mn(12)-Ac), which also crystallizes in an axial (tetragonal) space group but which now is recognized to consist of a mixture of six hydrogen-bonded isomers in the crystal and thus gives multiple, inhomogeneously broadened EPR signals. Similarly, single crystal (55)Mn NMR spectra on Mn(12)-BrAc display much sharper signals than a single crystal of Mn(12)-Ac, and this allows one Mn(III) signal to show an almost baseline-resolved quintet from quadrupolar splitting ((55)Mn, I = 5/2, 100%), allowing quadrupole coupling parameters (e(2)qQ) to be determined. In addition, it was found that crushing crystals of Mn(12)-BrAc into a microcrystalline powder causes severe broadening and shifts of the NMR resonances, emphasizing the superiority of single-crystal studies. The combined results establish that Mn(12) BrAc is far superior to Mn(12)-Ac for the study of the intrinsic properties of the Mn(12) family of SMMs in axial symmetry, and for the search for new phenomena such as quantum interference effects caused by higher-order (>2nd-order) transverse terms in the spin Hamiltonian. PMID- 16719479 TI - Nanosized polymetallic resorcinarene-based host assemblies that strongly bind fullerenes. AB - Polymetallic nanodimensional assemblies have been prepared via metal directed assembly of dithiocarbamate functionalized cavitand structural frameworks with late transition metals (Ni, Pd, Cu, Au, Zn, and Cd). The coordination geometry about the metal centers is shown to dictate the architecture adopted. X-ray crystallographic studies confirm that square planar coordination geometries result in "cagelike" octanuclear complexes, whereas square-based pyramidal metal geometries favor hexanuclear "molecular loop" structures. Both classes of complex are sterically and electronically complementary to the fullerenes (C(60) and C(70)). The strong binding of these guests occurred via favorable interactions with the sulfur atoms of multiple dithiocarbamate moieties of the hosts. In the case of the tetrameric copper(II) complexes, the lability of the copper(II) dithiocarbamate bond enabled the fullerene guests to be encapsulated in the electron-rich cavity of the host, over time. The examination of the binding of fullerenes has been undertaken using spectroscopic and electrochemical methods, electrospray mass spectrometry, and molecular modeling. PMID- 16719481 TI - Compositional dependence of negative thermal expansion in the Prussian Blue analogues M(II)Pt(IV)(CN)6 (M = Mn, Fe, Co, Ni, Cu, Zn, Cd). AB - The effect of M(II) substitution on the magnitude of the negative thermal expansion (NTE) behavior within a series of Prussian Blue analogues, M(II)Pt(IV)(CN)(6) for M(II) = Mn, Fe, Co, Ni, Cu, Zn, Cd, has been investigated using variable-temperature powder X-ray diffraction (100-400 K). The NTE behavior varies widely with M(II) substitution, from near zero thermal expansion in NiPt(CN)(6) (alpha = dl/l dT = -1.02(11) x 10(-)(6) K(-)(1)) up to a maximum in CdPt(CN)(6) (alpha = -10.02(11) x 10(-)(6) K(-)(1)). The trend in the magnitude of the NTE behavior, with increasing atomic number (Z) of the M(II) ion, follows the order Mn(II) > Fe(II) > Co(II) > Ni(II) < Cu(II) < Zn(II) < Cd(II), which correlates with the trends for M(II) cation size, the lattice parameter, and structural flexibility as indicated by the temperature-dependent structural refinements and Raman spectroscopy. Analysis of the temperature dependence of the average structures suggests that the differences in the thermal expansion are due principally to the different strengths of the metal-cyanide binding interaction and, accordingly, the different energies of transverse vibration of the cyanide bridge, with enhanced NTE behavior for more flexible lattices. PMID- 16719480 TI - Selective DNA strand scission with binuclear copper complexes: implications for an active Cu2-O2 species. AB - A homologous series of binuclear copper(II) complexes [Cu(II)(2)(Nn)(Y)(2)](2+) (1-3) (n = 3-5 and Y = (ClO(4))(-) or (NO(3))(-)) were studied to investigate the intermediate(s) responsible for selective DNA strand scission in the presence of MPA/O(2) (MPA = 3-mercaptopropanoic acid). While the N3 complex does not react, the N4 and N5 analogues show comparable activity with strand scission occurring at a single-strand/double-strand junction. Identical reactivity is also observed in the alternate presence of H(2)O(2). Spectroscopic and reactivity studies with [Cu(II)(2)(N4)(Y)(2)](2+) (2) and H(2)O(2) are consistent with DNA oxidation mediated by formation of a side-on peroxodicopper(II) (Cu(2)-O(2)) complex. PMID- 16719483 TI - Convergent, enantioselective syntheses of guanacastepenes A and E featuring a selective cyclobutane fragmentation. AB - The evolution of a convergent strategy that led to efficient, enantioselective syntheses of both natural (+)- and unnatural (-)-guanacastepene E and formal total syntheses of (+)- and (-)-guanacastepene A is described. A union of five- and six-membered ring intermediates by an efficient pi-allyl Stille cross coupling reaction was followed by an intramolecular enone-olefin [2 + 2] photocycloaddition and a stereoelectronically controlled, reductive fragmentation of the resulting cyclobutyl ketone. The latter two transformations enabled controlled formation of the C-11 quaternary stereocenter and the central seven membered ring of the guanacastepenes. An enantiospecific synthesis of the functionalized five-membered ring vinyl stannane from the monoterpene R-(-) carvone featuring a carbon-carbon bond forming ring contraction was also developed. PMID- 16719482 TI - Polymorphism of the signaling molecule c-di-GMP. AB - Using UV, CD, and NMR, we demonstrate that the important bacterial signaling molecule involved in biofilm formation, cyclic diguanosine monophosphate (c-di GMP), exists as a mixture of five different but related structures in an equilibrium that is sensitive both to its concentration and to the metal present. At the lower concentrations used for UV and CD work (0.05-0.5 mM), Li(+), Na(+), Cs(+), and Mg(2+) favor a bimolecular self-intercalated structure, while K(+), Rb(+), and NH(4)(+) favor formation of one or more guanine quartet complexes as well. At the higher NMR concentrations ( approximately 30 mM), the bimolecular structures associate and rearrange to a mixture of all-syn and all-anti tetramolecular and octamolecular quartet complexes. With K(+) the octamolecular complexes predominate, while with Li(+) the tetramolecular and octamolecular quartet complexes are present in approximately equal amounts, along with the bimolecular structure. We also find that both guanine amino groups in c-di-GMP are essential for formation of the quartets, because substitution of inosine for one guanosine allows formation of only the bimolecular structure. Further, two molecules of c-di-GMP tethered together are constrained in such a way that limits their ability to form these quartet complexes. The polymorphism we describe may provide different options for this signaling molecule when performing its functions in a bacterial cell, with K(+) and its own local concentration controlling the equilibrium. PMID- 16719484 TI - Spontaneous CdTe --> alloy --> CdS transition of stabilizer-depleted CdTe nanoparticles induced by EDTA. AB - CdTe nanoparticles stabilized by l-cysteine are chemically transformed into CdS nanoparticles of the same diameter via an intermediate CdTeS alloy without any auxiliary source of sulfur. The reaction is induced by ethylenediaminetetraacetic acid dipotassium salt dehydrate (EDTA), which was demonstrated experimentally to act as a catalyst by partially removing thiol stabilizers from the nanoparticle surface. It is hypothesized that addition of EDTA facilitates Te(2-) release, and oxidation of Te(2-) drives the nanoparticle transition process. Unlike many reports on reactions catalyzed by nanocolloids, this is likely to be the first observation of a catalytic reaction in which nanoparticles function as a substrate rather than a catalyst. It opens new pathways for the synthesis of novel nanoscale II-VI and other semiconductors and represents an interesting case of chemical processes in nanocolloids with reactivity increased by depletion of the surface layer of thiol stabilizers. This includes but is not limited to accurate control over the particle composition and crystallization rate. The slow rate of the CdTe --> alloy --> CdS transition is important for minimizing defects in the crystal lattice and results in a substantial increase of the quantum yield of photoluminescence over the course of the transition. PMID- 16719485 TI - Unidirectional electronic ring current driven by a few cycle circularly polarized laser pulse: quantum model simulations for Mg-porphyrin. AB - A circularly polarized ultraviolet (UV) laser pulse may excite a unidirectional valence-type electronic ring current in an oriented molecule, within the pulse duration of a few femtoseconds (e.g., tau = 3.5 fs). The mechanism is demonstrated by quantum model simulation for |X = |1(1)A(1g) --> |E(+) = |4 (1)E(u+) population transfer in the model system, Mg-porphyrin. The net ring current generated by the laser pulse (I = 84.5 microA) is at least 100 times stronger than any ring current, which could be induced by means of permanent magnetic fields, with present technology. PMID- 16719486 TI - Highly chemo-, regio-, and stereoselective [3+2]-cyclization of activated and deactivated allenes with alkenyl Fischer carbene complexes: a straightforward access to alkylidenecyclopentanone derivatives. AB - A broad range of functionalized 5-alkylidenecyclopentene derivatives are synthesized by the rhodium(I)-catalyzed [3+2]-cyclization reaction of chromium alkenyl(methoxy)carbene complexes 1 and activated allenes. Thus, amidocyclopentenes 4a-n are readily available from N-allenylamides 2a-c, while phenoxyallene 2e gives access to phenoxycyclopentenes 6. In turn, the cyclization reaction with (alkoxycarbonyl)allenes 3 leads to (alkoxycarbonyl)methylidenecyclopentenes 7-10. In terms of selectivity, most cyclization reactions take place with complete chemo-, regio-, and diastereoselectivity. Representative cycloadducts are efficiently hydrolyzed to the corresponding 2-alkylidenecyclopentanones 11a-e without tautomerization or isomerization. Finally, a tentative reaction pathway is proposed that involves the rhodium(I) carbene complexes as the species responsible for the [3+2] cyclization. PMID- 16719487 TI - Synthesis and electronic factors in thermal cyclodimerization of functionalized aromatic trifluorovinyl ethers. AB - A series of 19 p-substituted aromatic trifluorovinyl ether compounds were prepared from versatile intermediate p-Br-C(6)H(4)-O-CF=CF(2) and underwent thermal radical mediated cyclodimerization to new difunctional compounds containing the 1,2-disubstituted perfluorocyclobutyl (PFCB) linkage. The synthetic scope demonstrates the functional group transformation tolerance of the fluorovinyl ether, and the dimers are useful as monomers for traditional step growth polymerization methods. (19)F NMR spectra confirmed that p-substitution affects the trifluorovinyl ether group chemical shifts. The first kinetic studies and substituent effects on thermal cyclodimerization were performed, and the results indicated that electron-withdrawing groups slow the rate of cyclodimerization. The data were further analyzed using the Hammett equation, and reaction constants (rho) of -0.46 at 120 degrees C and -0.59 at 130 degrees C were calculated. This study presents the first liner free energy relationship reported for the cyclodimerization of aromatic trifluorovinyl ethers to PFCB compounds. PMID- 16719488 TI - Hydrogenation of arenes over silica-supported catalysts that combine a grafted rhodium complex and palladium nanoparticles: evidence for substrate activation on Rh(single-site)-Pd(metal) moieties. AB - The complex Rh(cod)(sulfos) (Rh(I); sulfos = ( )O(3)S(C(6)H(4))CH(2)C(CH(2)PPh(2))(3); cod = cycloocta-1,5-diene), either free or supported on silica, does not catalyze the hydrogenation of benzene in either homogeneous or heterogeneous phase. However, when silica contains supported Pd metal nanoparticles (Pd(0)/SiO(2)), a hybrid catalyst (Rh(I)-Pd(0)/SiO(2)) is formed that hydrogenates benzene 4 times faster than does Pd(0)/SiO(2) alone. EXAFS and DRIFT measurements of in situ and ex situ prepared samples, batch catalytic reactions under different conditions, deuterium labeling experiments, and model organometallic studies, taken together, have shown that the rhodium single sites and the palladium nanoparticles cooperate with each other in promoting the hydrogenation of benzene through the formation of a unique entity throughout the catalytic cycle. Besides decreasing the extent of cyclohexa-1,3 diene disproportionation at palladium, the combined action of the two metals activates the arene so as to allow the rhodium sites to enter the catalytic cycle and speed up the overall hydrogenation process by rapidly reducing benzene to cyclohexa-1,3-diene. PMID- 16719489 TI - Reversible carbon-carbon bond formation between 1,3-dienes and aldehyde or ketone on nickel(0). AB - The reversible oxidative cyclization of dienes and aldehydes with nickel(0) proceeded to give eta(3):eta(1)-allylalkoxynickel complexes. The treatment of these complexes with carbon monoxide led to the formation of the corresponding lactone and/or the regeneration of a butadiene and an aldehyde concomitant with the formation of Ni(CO)(3)(PCy(3)). The scission of the nickel-oxygen bond of the allylalkoxy complexes with ZnMe(2) leading to eta(3)-allyl(methyl)nickel was very efficient to suppress the reverse reaction of the oxidative cyclization. The methylated eta(3)-allylnickel compound underwent the reductive elimination. The carbonylative coupling reaction of the eta(3)-allyl(methyl)nickel proceeded as well under a carbon monoxide atmosphere. Similarly, the addition of Me(3)SiCl to eta(3):eta(1)-allylalkoxynickel complexes was also efficient for the inhibition of the reverse reaction. The resulting eta(3)-1-siloxyethylallylnickel complex was treated with carbon monoxides followed by the addition of MeOH to give the expected hydroxyester. This method is efficient as well even for the eta(3):eta(1)-allyl(alkoxy)nickel complex containing acetone as a component, which was so prone to undergo the reverse reaction hampering its isolation. The isolation of the eta(3):eta(1)-allylalkoxynickel complex containing ketone as a component was made easier by the use of heavier butadiene and ketone, such as 2,3 dibenzyl-1,3-butadiene and benzophenone or by the use of cyclobutanone. The reaction with styrene oxide gave the eta(3):eta(1)-allylalkoxynickel containing phenylacetoaldehyde, an isomer of styrene oxide. PMID- 16719490 TI - A low-temperature synthesis for organically soluble HgTe nanocrystals exhibiting near-infrared photoluminescence and quantum confinement. AB - A new low-temperature, one-pot method is introduced for the preparation of organically passivated HgTe nanocrystals, without the use of highly toxic precursors. The nanocrystals show bright photoluminescence in the infrared telecommunication windows about 1300 and 1550 nm with quantum efficiencies between 55 and 60%. They have a zinc blende structure with a mean particle diameter of 3.4 nm, thus exhibiting quantum confinement effects. Particle growth is self-limited by temperature quenching, so a narrow size distribution is obtained. The measured size of the particles agrees with calculations using the pseudopotential method. PMID- 16719491 TI - Formation and decomposition of C3 metallacycles from ethylene and methylene on MoAl alloy thin films. AB - The reaction between adsorbed ethylene and methylene species has been investigated on a molybdenum-aluminum alloy grown from Mo(CO)(6) on a planar alumina film formed on a Mo(100) single crystal in ultrahigh vacuum. Di-sigma bonded ethylene reacts with carbene species, formed on the surface from methylene iodide, to form a C(3) metallacycle. This predominantly decomposes to yield propylene, while a smaller portion yields cross-metathesis products since (12)C(13)CH(4) is formed from reaction between (13)C(2)H(4) and (12)CH(2). This work demonstrates for the first time that the reaction proceeds in heterogeneous phase via a C(3) metallacycle as proposed in the Herisson-Chauvin mechanism. PMID- 16719492 TI - Reversible photoswitching of the coordination numbers of silicon in organosilicon compounds bearing a 2-(phenylazo)phenyl group. AB - Several organosilicon compounds bearing a 2-(phenylazo)phenyl group were synthesized from the corresponding chlorosilanes and 2-lithioazobenzene prepared by halogen-lithium transmetalation of 2-iodoazobenzene. Their structures were determined by (1)H, (13)C, (19)F, and (29)Si NMR spectra, UV-vis spectra, and X ray crystallographic analyses. In the UV-vis spectra, silyl groups caused red shifts of both the n-pi and pi-pi transitions of the azo group compared with the transitions of the unsubstituted azobenzene. The E-isomers of the fluorosilanes showed an intramolecular interaction between a nitrogen atom of the azo group and the silicon atom, leading their intermediate structures between a distorted trigonal bipyramidal structure and a tetrahedral structure around the silicon atoms, which were revealed by the X-ray crystallographic analyses and the NMR spectra. On the other hand, silanes without fluorine atoms showed tetrahedral structures in the absence of such an interaction. The photoirradiation of the E isomers of the fluorosilanes afforded reversibly the corresponding Z-isomers in good yields. The silicon atoms of the Z-isomers were found to be tetracoordinate in the absence of Si-N interactions by the (29)Si NMR spectra. The coordination numbers of the silicon atom of the fluorosilanes were reversibly switched between four and five by photoirradiation. These properties were compared to those of a tetrafluoro[2-(phenylazo)phenyl]silicate. PMID- 16719493 TI - U1 and U2 small nuclear RNA genetic linkage: a novel molecular tool for identification of six sole species (Soleidae, Pleuronectiformes). AB - We evaluated the usefulness of a genetic linkage between the U1 and U2 small nuclear RNAs for species identification. Six soles belonging to the genera Solea, Dicologlossa, and Microchirus were studied. A simple methodology based on two single PCRs is described. Reproducible band profiles were generated for all samples. This rapid and discriminatory molecular method is highly promising for determining the authenticity of sole fillets in the food industry. PMID- 16719494 TI - Development and validation of an HPLC/UV/MS method for simultaneous determination of 18 preservatives in grapefruit seed extract. AB - Grapefruit seed extracts are used in cosmetics, food supplements, and pesticides because of their antimicrobial properties, but suspicions about the true nature of the active compounds arose when synthetic disinfectants such as benzethonium or benzalkonium chloride were found in commercial products. The HPLC method presented herein allows the quality assessment (qualitative and quantitative) of these products for the first time. On the basis of a standard mixture of 18 preservatives most relevant for food and grapefruit products, a method was developed allowing the baseline separation of all compounds within 40 min. Optimum results were obtained with a C-8 stationary phase and a solvent system comprising aqueous trifluoroacetic acid, acetonitrile, and 2-propanol. The assay was fully validated and shown to be sensitive (LOD < or= 12.1 ng on-column), accurate (recovery rates > or = 96.1%), repeatable (sigma(rel) < or = 3.5%), precise (intra-day variation < or = 4.5%, interday variation < or = 4.1%), and rugged. Without any modifications the method could be adopted for LC-MS experiments, where the compounds of interest were directly assignable in positive ESI mode. The quantitative results of several products for ecofarming confirmed previous studies, as seven out of nine specimens were adulterated with preservatives in varying composition. The samples either contained benzethonium chloride (2.5-176.9 mg/mL) or benzalkonium chloride (138.2-236.3 mg/mL), together with smaller amounts of 4-hydroxybenzoic acid esters, benzoic acid, and salicylic acid. PMID- 16719495 TI - Chemical compositions, antioxidant capacities, and antiproliferative activities of selected fruit seed flours. AB - Seed flours from black raspberry, red raspberry, blueberry, cranberry, pinot noir grape, and chardonnay grape were examined for their total fat content, fatty acid composition, total phenolic content (TPC), total anthocyanin content (TAC), radical scavenging capacities against the peroxyl (ORAC) and stable DPPH radicals, chelating capacity against Fe(2+), and antiproliferative activities using the HT-29 colon cancer cell line. Significant levels of fat were detected in the fruit seed flours and their fatty acid profiles may differ from those of the respective seed oils. Cranberry seed flour had the highest level of alpha linolenic acid (30.9 g/100 g fat) and the lowest ratio of n-6/n-3 fatty acids (1.2/1). The ORAC value of the chardonnay seed flour was 1076.4 Trolox equivalents mumol/g flour, and its TPC was 186.3 mg gallic acid equivalents/g flour. These values were 3-12 times higher than the other tested fruit seed flours. Furthermore, the ORAC value was significantly correlated to the TPC under the experimental conditions (P < 0.05). These fruit seed flours also differed in their TAC values and Fe(2+)-chelating capacities. In addition, black raspberry, cranberry, and chardonnay grape seed flour extracts were evaluated for their antiproliferative effects using HT-29 colon cancer cells. All three tested seed flour extracts significant inhibited HT-29 cell proliferation. The data from this study suggest the potential of developing the value-added use of these fruit seed flours as dietary sources of natural antioxidants and antiproliferative agents for optimal human health. PMID- 16719497 TI - Isolation and characterization of rosmarinic acid oligomers in Celastrus hindsii Benth leaves and their antioxidative activity. AB - Antioxidative compounds were isolated from the 50% methanol extract of dried leaves of Celastrus hindsii. Eight phenolic compounds (1-8) were finally obtained by reversed-phase high-performance liquid chromatography, and their structures were elucidated by nuclear magnetic resonance spectrometry and mass spectrometry analyses. They were the five known compounds, rutin (1), kaempferol 3-rutinoside (2), rosmarinic acid (3), lithospermic acid (4), and lithospermic acid B (6), and three novel oligomers of rosmarinic acid, a dimer (5) and two trimers (7 and 8). The major components in the extract were rosmarinic acid (3) and lithospermic acid B (6). These phenolic compounds were shown to have antioxidative activities against the autoxidation of methyl linoleate in bulk phase and the radical initiated peroxidation of soybean phosphatidylcholine in liposomes. In the liposomal peroxidation, the number of phenolic hydroxyl group in each molecule was correlated with the effectiveness of antioxidative activity. PMID- 16719496 TI - Lipophilic hydroxytyrosyl esters. Antioxidant activity in lipid matrices and biological systems. AB - Antioxidant activities of lipophilic hydroxytyrosyl acetate, palmitate, oleate, and linoleate were compared with those of hydroxytyrosol, alpha-tocopherol, and butylhydroxytoluene (BHT) in both glyceridic matrix and biological systems. Aliquots of a glyceridic matrix spiked with various concentrations of antioxidant were subjected to accelerated oxidation in a Rancimat apparatus operated at 90 degrees C. The relationships between induction time (IT) and antioxidant concentration (mmol/kg) presented by hydroxytyrosol and hydroxytyrosyl acetate, palmitate, oleate, and linoleate were similar. Hydroxytyrosol and its esters showed greater antioxidant activity than alpha-tocopherol or BHT. We also evaluated the capacity of hydroxytyrosyl esters to protect proteins and lipids against oxidation caused by peroxyl radicals, using a brain homogenate as an ex vivo model. All tested compounds showed a protective effect in these systems, which was greater in preventing the generation of carbonyl groups in protein than of malondialdehyde in lipid. Inclusion of a lipophilic chain in the hydroxytyrosol molecule enhanced its antioxidant capacities in this biological model. PMID- 16719498 TI - Antioxidant potentials of flaxseed by in vivo model. AB - The present study reports the antioxidant activity of flaxseed as measured by feeding weanling albino rats with 5.0% and 10.0% of flaxseed (constituting approximately 0.75 and 1.5 g kg(-)(1)) for 14 days followed by challenging animals with 2.0 g kg(-)(1) b.w. CCl(4) as toxin. Activity was assessed by measuring hepatic marker enzymes like catalase, superoxide dismutase (SOD), and peroxidase and comparing with those from the normal group and from a group receiving toxin without flaxseed. Treatment of CCl(4) at dose of 2.0 g kg(-)(1) b.w. decreased the activities of various antioxidant enzymes such as catalase, superoxide dismutase (SOD), and peroxidase by 35.6%, 47.76%, and 53.0%, respectively, compared to the control group, and the lipid peroxidation value increased nearly 1.2-fold compared to that of the group treated with toxin without flaxseed. Pretreatment of rats with 5.0% flaxseed followed by CCl(4) treatment caused restoration of catalase, SOD, and peroxidase by 39.7%, 181.42%, and 123.7%, respectively, as compared to control. The group treated with 10.0% flaxseed has shown the restoration of 95.02%, 182.31%, and 136.0% of catalase, SOD, and peroxidase. In the case of the group treated with toxin without flaxseed, the level of superoxide dismutase and the catalse value decreased 91.4% and 55.33%, respectively, in comparison with the control group. These results clearly indicate the beneficial effect of flaxseed components as an antioxidant as seen by restoration of hepatic enzymes, which were varied from normal to one due to toxicity induced by toxin (CCl(4)). Owing to this property, the flaxseed known for its functional properties can be further extended to exploit its possible application for various health benefits as nutraceuticals and food ingredient. PMID- 16719499 TI - Inhibition of bacteriophage m13 replication with esterified milk proteins. AB - Esterified milk proteins [methylated (Met) or ethylated (Et) alpha-lactalbumin (ALA), beta-lactoglobulin (BLG), and beta-casein (BCN)], unmodified native milk proteins, and native basic proteins (calf thymus histone and hen egg white lysozyme) were tested for their antiviral activity against the bacteriophage M13 and for their influence on its replication (except BCN). All esterified milk proteins showed an antiviral activity against the bacteriophage M13, proportional to the extent of esterification and, hence, to the increased basicity of the modified proteins. Antiviral activity of 100% Met-BLG disappeared after its pepsinolysis but not after its trypsinolysis. The antiviral activity of Met-BLG was much higher than that of native basic proteins (histone and lysozyme). One hundred percent Met-BLG and 73% Et-BLG inhibited the replication of bacteriophage M13 completely, whereas 60% Met-ALA inhibited phage replication partially. Calf thymus histone inhibited the replication of bacteriophage M13 at a lower extent (20%) than Met- and Et-BLG (100% inhibition). Protein concentration, pH, and concentration of the Escherichia coli culture in the preincubation medium of the virus were other factors influencing antiviral activity. Interactions of esterified proteins with the phage DNA (phenol extracted) followed the same pattern as observed during studies of the inhibition of the phage replication: Met-BLG > Et-BLG > or = Met-ALA. PMID- 16719500 TI - Bioactive compounds in legumes and their germinated products. AB - Nineteen domestic legume varieties, including 6 soybeans, 7 black soybeans, 4 azuki beans, and 2 mung beans, were evaluated for contents of dietary fiber, total phenolics, and flavonoids. Nine varieties of legumes (black soybean TN6, TN3, BM, and WY; soybean KS1, KS2, and KS8; azuki bean AKS5 and AKS6) were good sources of bioactive compounds and were selected for germination tests. After short- and long-term germinations, the bioactive compounds were determined and compared with compositions of isoflavones in soybeans. The reducing power of legumes correlated well with their total flavonoid contents (r (2) = 0.9414), whereas less correlation was found between reducing power and total phenolics contents (r (2) = 0.6885). The dark-coat seeds, such as azuki beans and black soybeans, contained high amounts of phenolic compounds and contributed to high antioxidative ability, whereas their phenolics content and antioxidative abilities significantly decreased after short-term germination due to losses of pigments in the seed coats. After long-term germination, the contents of bioactive compounds (total phenolics and flavonoids) increased again and the ratio of aglycones to total isoflavones significantly increased in black soybeans. TN3 and TN6 seeds and their long-term germinated seeds and AKS5 seeds were identified as the legume samples that might have the highest antioxidant ability according to the results of chemometric analysis. Selection of the right legume varieties combined with a suitable germination process could provide good sources of bioactive compounds from legumes and their germinated products for neutraceutical applications. PMID- 16719501 TI - Structural and biological characterization of sulfated-derivatized oat beta glucan. AB - The structural, physicochemical, and biological properties of sulfated oat beta glucan were characterized. The degree of substitution of the sulfated oat-beta glucan was obtained by elemental analysis, which was 0.68. Compared to native oat beta-glucan, the FT-IR spectra of the derivative showed two new absorption bands at 1250 and 810 cm(-)(1), which would be attributed to (S=O) and (C-O-S) groups, respectively. The molecular weight of the sulfated beta-glucan was determined to be 68 kDa and its viscosity decreased by almost 2 orders of magnitude while its solubility increased by more than 100% compared to that of the native beta glucan. In addition, the sulfation caused the reduction of in vitro bile acid binding capacity of oat beta-glucan due to the new anionic character and decreased molecular weight. The sulfated derivative exhibited, however, anticoagulant activity which showed a concentration-dependent increase. PMID- 16719502 TI - Water-soluble genistin glycoside isoflavones up-regulate antioxidant metallothionein expression and scavenge free radicals. AB - Genistin has antioxidant activities; however, its insolubility in water often limits its biological availability in vivo. Using a novel transglycosylation process, the solubility of genistin glycosides was increased 1000 to 10000-fold, but it was not known whether these modified genistin glycosides maintained antioxidant activity. We found that both genistin and its glycosides similarly up regulated the transcription of several metallothionein (MT) antioxidant genes (MT1A, MT2A, MT1E, and MT1X), as well as the glucose 6-phosphate dehydrogenase (G6PD) gene in HepG2 cells. This gene induction was mediated by the sequestration of zinc in the cytosol, which up-regulated the metal-responsive transcription factor-1 (MTF-1) that induced MT gene expression. Although not as effective as ascorbic acid, genistin glycosides possessed slightly greater reducing power than genistin. We concluded that genistin and genistin glycosides have a direct antioxidant effect and an indirect antioxidant effect, perhaps via induction of MT by activity of MTF-1. PMID- 16719503 TI - Evaluation of the antioxidant capacity of limonin, nomilin, and limonin glucoside. AB - The antioxidant capacity (AOC) of three representative citrus limonoids, limonin, nomilin, and limonin glucoside, was examined by the oxygen radical absorbance capacity (ORAC), Trolox equivalent antioxidant capacity (TEAC), beta-carotene linoleic acid bleaching, and 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging assays. Pure compounds and proper negative (cinnamic acid) and positive (2,6-di-tert-butyl-4-methylphenol (BHT) and ascorbic acid) controls were used to remove any ambiguity in interpreting results. In all cases, limonin and nomilin gave results equivalent to those of cinnamic acid, indicating that they do not possess any inherent AOC and should not be considered antioxidants. Similar results were observed for limonin glucoside, with the exception of an anomalous result obtained from the beta-carotene-linoleic acid bleaching assay. Limonin glucoside was deemed not to be an antioxidant on the basis of the three unequivocal assays. PMID- 16719504 TI - Influence of peptide supply and cosubstrates on phenylalanine metabolism of Lactobacillus sanfranciscensis DSM20451(T) and Lactobacillus plantarum TMW1.468. AB - Bread spoilage is mainly due to the growth of filamentous fungi, and metabolites produced during sourdough fermentation by lactobacilli can inhibit fungal growth. One of these metabolites is phenyllactic acid (PLA), which is a catabolite from phenylalanine. In this work, the influence of peptide supply and cosubstrates was determined on PLA formation from phenylalanine by Lactobacillus plantarum TMW1.468 and Lactobacillus sanfranciscensis DSM20451(T). Transport of single amino acids is not efficient in lactobacilli, and only 1% of the offered phenylalanine was converted to PLA. PLA yields were increased 2-4-fold when peptides instead of single amino acids were used as a substrate. The accumulation of phenylalanine after peptide addition indicated that, after transport, transamination was the second limiting factor. In L. plantarum TMW1.468, PLA yields were increased from 5 to >30% upon the addition of alpha-ketoglutarate. In L. sanfranciscensis DSM20451, a combination of both citric acid and alpha ketoglutarate increased PLA formation. The combined effect of citric acid and alpha-ketoglutarate can be attributed to changes in the NAD/NADH ratio. PMID- 16719505 TI - Lactobacillus plantarum and Pediococcus pentosaceus starter cultures as a tool for microflora management in malting and for enhancement of malt processability. AB - Lactobacillus plantarum VTT E-78076 (E76) and Pediococcus pentosaceus VTT E-90390 (E390) starter cultures were added to the steeping water of normal malting barley in order to balance the microbial community and to enhance malt processability. In this study, we also investigated the effects of lactic acid-acidified MRS spent medium (MRS-LA) on malting performance. Malting trials with five different two-row barley varieties were carried out in 25 kg pilot scale. The starter cultures promoted yeast growth during malting and restricted the growth of harmful bacteria and Fusarium fungi. Furthermore, they had positive effects on malt characteristics. Reduction in wort viscosity and beta-glucan content and enhanced xylanase and microbial beta-glucanase activities were observed. Starter cultures notably improved lautering performance. Some of the beneficial effects were due to the lactic acid and low pH, as similar effects were obtained with MRS LA. Starter cultures offer a tool for tailoring of malt properties. PMID- 16719506 TI - New process for fungal delignification of sugar-cane bagasse and simultaneous production of laccase in a vapor phase bioreactor. AB - We propose a new process using a vapor phase bioreactor (VPB) to simultaneously (i) delignify sugar-cane bagasse, a residue of sugar production that can be recycled in paper industry, and (ii) produce laccase, an enzyme usable to bleach paper pulp. Ethanol vapor, used as laccase inducer, was blown up through a VPB packed with bagasse and inoculated with Pycnoporus cinnabarinusss3, a laccase hyperproducing fungal strain. After 28 days, the laccase activity in the ethanol treated bagasse was 80-fold higher (80 U g(ds)(-)(1)) and the bagasse delignification percentage was 12-fold (12%) higher than in the reference samples produced in the absence of ethanol, corresponding to a high overall pulp yield of 96.1%. In the presence of ethanol, the total soluble phenols amount was 2.5-fold (3 mg FA g(ds)(-)(1)) higher than that without ethanol. Six monomeric phenols were detected: p-coumaric (4-hydroxyphenyl-2-propenoic), ferulic (4-hydroxy-3 methoxyphenyl-2-propenoic), syringic (4-hydroxy-3,5-dimethoxybenzoic), vanillic (4-hydroxy-3-methoxybenzoic) and 4-hydroxybenzoic acids, and 2 methoxyhydroquinone. Higher concentrations of phenolic compounds were observed when ethanol vapor was added, confirming a more efficient bagasse delignification. After 28 days, the fungal-treated bagasse (with ethanol addition) was pulped and refined. For a freeness of 81 mL CSF, this processing required 50% less energy than with untreated bagasse (without inoculation and ethanol addition), which indicated a significant potential economy for the pulp and paper industry. Handsheets were made from pulp obtained after fungal-treated and untreated bagasse. Comparison of bagasse-pulp characteristics for freeness of 35 and 181 mL CSF showed an average increment by 35% for tensile index and breaking strength and length. VPB allowed a simultaneous production of laccase (90 U g(ds)(-)(1), after pressing of the bagasse) that improved the overall profitability of the process. PMID- 16719507 TI - Genomic organization of a diverse ACC synthase gene family in banana and expression characteristics of the gene member involved in ripening of banana fruits. AB - The banana is one of the typical climacteric fruits with great economic importance in agriculture. To understand the basic mechanism underlying banana ripening, gene clones for banana ACC synthase (EC 4.4.1.14), a key regulatory enzyme in the ethylene biosynthetic pathway, were characterized. Genomic clones were analyzed by restriction mapping, and the data in conjunction with sequence comparisons with the previously isolated PCR fragments indicated that at least nine ACC synthase genes (MACS1-9) exist in the banana genome. Southern blot analysis showed they are located in different regions of the banana genome. Three lambda genomic clones (GMACS-1, -9, and -12) were completely sequenced, and gene structures of MACS1 (corresponding to alleles of GMACS-9 and -12) and MACS2 (corresponding to GMACS-1) were elucidated. The coding regions of these three genes were all interrupted by three introns. The size and location of introns are similar to the ACC synthase genes from tomato and Arabidopsis. Northern analysis showed that only MACS1 expressed during fruit ripening and was inducible by exogenous ethylene treatment, which indicates MACS1 is a significant member of the ACC synthase gene family related to ripening in banana fruit. The transcription initiation site of GMACS-12 containing MACS1 was defined. There is a TATTAAT sequence located at position -31 to -25 that qualifies as a TATA box. The delineation of transcription unit in MACS1 will facilitate the promoter studies for this gene and allow its specific functions involved in fruit ripening to be determined. PMID- 16719508 TI - The use of Lactobacillus pentosus 1MO to shorten the debittering process time of black table olives (Cv. Itrana and Leccino): a pilot-scale application. AB - Fifty lactobacilli isolated from black table olive brines were evaluated for their salt tolerance, resistance to oleuropein and verbascoside, and ability to grow in modified filter-sterilized brines. A strain of Lactobacillus pentosus was selected and used as a starter to ferment, in pilot plant, black olives (Itrana and Leccino cv.) in brines modified for pH, carbohydrate, and growth factor concentrations, at 28 degrees C. The temperature-controlled fermentation of Leccino cv. olives resulted in obtaining ready-to-eat, high-quality table olives in a reduced-time process. HPLC analysis of phenolic compounds from fermented olives showed a decrease of oleuropein, a glucoside secoiridoid responsible for the bitter taste of olive drupes, and an increase of the hydroxytyrosol concentration. The selected strain of L. pentosus (1MO) allowed the reduction of the debittering phase period to 8 days. PMID- 16719509 TI - Interactions between yeast lees and wine polyphenols during simulation of wine aging. II. Analysis of desorbed polyphenol compounds from yeast lees. AB - In the first part of this work, the analysis of the polyphenolic compounds remaining in the wine after different contact times with yeast lees during simulation of red wine aging was undertaken. To achieve a more precise view of the wine polyphenols adsorbed on lees during red wine aging and to establish a clear balance between adsorbed and remnant polyphenol compounds, the specific analysis of the chemical composition of the adsorbed polyphenolic compounds (condensed tannins and anthocyanins) after their partial desorbtion from yeast lees by denaturation treatments was realized in the second part of the study. The total recovery of polyphenol compounds from yeast lees was not complete, since a rather important part of the initial wine colored polyphenols, especially those with a dominant blue color component, remained strongly adsorbed on yeast lees, as monitored by color tristimulus and reflectance spectra measurements. All anthocyanins were recovered at a rather high percentage (about 62%), and it was demonstrated that they were not adsorbed in relation with their sole polarity. Very few monomeric phenolic compounds were extracted from yeast lees. With the use of drastic denaturing treatments, the total recovery of condensed tannins reached 83%. Such tannins extracted from yeast lees exhibited very high polymeric size and a rather high percentage of galloylated residues by comparison with initial wine tannins, indicating that nonpolar tannins were preferentially desorbed from yeast lees by the extraction treatments. PMID- 16719510 TI - Use of Aloe vera gel coating preserves the functional properties of table grapes. AB - Table grapes (Vitis vinifera L. cv. Crimson Seedless) were coated with Aloe vera gel according to our developed patent (SP Patent P200302937) and then stored for 35 days at 1 degrees C, and the subsequent shelf life (SL) was monitored at 20 degrees C. Uncoated clusters showed a rapid loss of functional compounds, such as total phenolics and ascorbic acid. These changes were accompanied by reduction of the total antioxidant activity (TAA) and increases in total anthocyanins, showing an accelerated ripening process. On the contrary, table grapes coated with Aloe vera gel significantly delayed the above changes, such as the retention of ascorbic acid during cold storage or SL. Consequently, Aloe vera gel coating, a simple and noncontaminating treatment, maintained the functional properties during postharvest storage of table grapes. PMID- 16719511 TI - Salicylic acid, ethephon, and methyl jasmonate enhance ester regeneration in 1 MCP-treated apple fruit after long-term cold storage. AB - Volatile esters, primarily synthesized in peel tissues, are major aromatic components of apple fruits [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.]. The use of cold storage combined with 1-methylcyclopropene (1-MCP) treatment prolongs the life of apples but represses the regeneration of esters during poststorage ripening. In this study, the regeneration of total esters was significantly increased in apple fruits treated with salicylic acid (SA) and Ethephon (ETH) that had been treated once or twice with 1-MCP. However, methyl jasmonate (MeJA) treatment resulted in regeneration of total esters after a single 1-MCP treatment. To determine the mechanism by which SA, ETH, and MeJA regulate ester regeneration, the apple alcohol acyltransferase gene (MdAAT2) was investigated at the mRNA, protein, and enzyme activity levels. Genes associated with ethylene perception were also investigated by RT-PCR. The results suggest that MdAAT2 controls ester regeneration and that MdETR1 plays a key role in ethylene perception and regulation of downstream MdAAT2 gene expression during poststorage. Ester compounds and concentrations differed in peels treated with different signal molecules, indicating that regulation of the pathway upstream of straight-chain ester biosynthesis depended on the regulation of lipoxygenase (LOX) and alcohol dehydrogenase (ADH) activity by SA, ETH, and MeJA during poststorage ripening. PMID- 16719512 TI - Cell wall composition in juvenile and adult leaves of maize (Zea mays L.). AB - Many leaf characteristics vary with position along the culm in maize (Zea mays L.) due to the existence of vegetative phase change and heteroblasty. The objective of this work was to determine if differences in cell wall composition exist among developmental phases and between Cg1, a developmental mutant, and wild-type maize. In one experiment, the middle third of fully elongated leaf blades from lower and upper regions of the shoot was harvested (midribs removed) and analyzed for several cell wall components. Averaged over five inbreds (De811, Ia5125, Mo17, P39, and Wh8584), lower leaf blades had higher levels of xylose and lower levels of total uronosyls, glucose, arabinose, and galactose (P < 0.05) than did upper leaf blades. With the exception of glucose, upper and lower leaves of Cg1 plants varied in the same manner as their near-isogenic siblings, except cell walls of Cg1 plants were more "juvenile" than cell walls of wild-type siblings at the same leaf stage. These data support the hypothesis that Cg1 delays but does not eliminate the transition from juvenile-vegetative to adult vegetative phase. In a second experiment, juvenile (leaves 3 and 5), transition (leaf 7), and adult (leaves 9 and 11) leaves from inbreds B73 and De811 were harvested and analyzed as in the first experiment. As leaf number rose, total cell wall content of sample dry matter, total neutral sugars, glucose, xylose, and ester-linked monomers of p-coumaric acid and total ferulates including ferulate dimers increased linearly while total uronosyls acids, arabinose, and galactose declined linearly (P < 0.05). Glucose and xylose are major cell wall components released from cellulose and xylans after acid hydrolysis. Pectin, a minor component of grass cell walls, is composed of galacturonosyls, arabinose, and galactose. Secondary cell wall deposition increased between leaves 3 and 11 in a heteroblastic series, due to either increased cell wall content concomitant with decreased cell lumen size, changes in proportion of cell types (i.e., sclerenchyma), or a combination of these factors. PMID- 16719513 TI - Proteomic analysis of hen egg white. AB - Hen egg white is an original biological fluid in which major proteins have been widely studied, unlike the minor components. In this study, two-dimensional electrophoresis associated with mass spectrometry enabled the separation of 69 protein spots and their matching with major proteins, which were already known, and with minor proteins. Sixteen proteins were identified, and among them, two had never been previously detected in hen egg white, i.e., Tenp, a protein with strong homology with a bacterial permeability-increasing protein family (BPI), and VMO-1, an outer layer vitelline membrane protein. Thirteen proteins present a very wide polymorphism (ovotransferrin, ovomucoid, clusterin, etc.), some of them up to nine isoforms (ovoinhibitor). Eleven functional protein families were identified (serpin, transferrin, protease inhibitors Kazal, glycosyl hydrolases, lipocalin, bactericidal permeability-increasing protein, clusterin, UPAR/CD59/Ly6/ snake neurotoxin, cysteine protease inhibitor, VMO-1, and folate receptor families). These various biological functions could be interesting for further valorizations. In addition, three spots remain unidentified, probably because these proteins are not yet indexed in the international protein databanks. PMID- 16719514 TI - Quantification of selected volatile constituents and anions in Mexican Agave spirits (Tequila, Mezcal, Sotol, Bacanora). AB - A large collection (n = 95) of Mexican Agave spirits with protected appellations of origin (Tequila, Mezcal, Sotol, and Bacanora) was analyzed using ion and gas chromatography. Because of their production from oxalate-containing plant material, all Agave spirits contained significant concentrations of oxalate (0.1 9.7 mg/L). The two Tequila categories ("100% Agave" and "mixed") showed differences in the methanol, 2-/3-methyl-1-butanol, and 2-phenylethanol concentrations with lower concentrations in the mixed category. Mezcal showed no significant differences in any of the evaluated parameters that would allow a classification. Sotol showed higher nitrate concentrations and lower 2-/3-methyl 1-butanol concentrations. Bacanora was characterized by exceptionally high acetaldehyde concentrations and a relatively low ethyl lactate content. The methanol content was the most problematic compound regarding the Mexican standards: two Tequilas (4%), five Sotols (31%), and six Bacanoras (46%) had levels above the maximum methanol content of 300 g/hL of alcohol. In conclusion, the composition of Mexican Agave spirits was found to vary over a relatively large range. PMID- 16719515 TI - Effect of six decades of selective breeding on soybean protein composition and quality: a biochemical and molecular analysis. AB - To evaluate the extent of the genetic change and its effects on the seed protein composition of soybean cultivars released during the past 60 years, representative ancestral cultivars and those derived from selective breeding were grown in a side-by-side comparison. Total seed protein content, determined by combustion analysis of nitrogen, revealed a decline in the protein content after decades of selection and breeding. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis comparison of protein profiles of the soybean cultivars indicated that relative expression of most of the seed storage proteins had not varied substantially from the ancestral lines to the present commercial cultivars. There was noticeably less beta-subunit of beta-conglycinin, a protein devoid of sulfur amino acids, in the modern cultivars represented by Mustang, Pioneer 93B09, and Asgrow 3602. Comparison of the amino acid profiles of soybean seed, a benchmark of the protein's nutritional quality, revealed that the ancestral progenitor, G. soja, was significantly higher in cysteine, glutamic acid, histidine, and arginine than either the ancestral or the modern cultivars. Selective breeding over the past 60 years minimally affected the overall amino acid composition. The degree of divergence in the DNA sequence of the genes encoding glycinin and beta conglycinin in the ancestral and modern cultivars was investigated using Southern hybridization and the polymerase chain reaction. Even though some restriction fragment polymorphisms could be detected, overall, the banding patterns were remarkably similar among the ancestral cultivars and those derived from them, suggesting a high degree of conservation of seed-storage protein genes. The results of our study suggest that selection and breeding for yield during the past 60 years had no major influence on the protein composition, ostensibly because of limited genetic diversity among the parental lines. PMID- 16719516 TI - Validation of a method to quantify copper and other metals in olive fruit by ETAAS. Application to the residual metal control after olive tree treatments with different copper formulations. AB - An electrothermal atomization atomic absorption spectrometry method was validated to quantify residues of copper, aluminum, cadmium, chromium, iron, lead, and nickel in olive fruit. The linearity ranges under the optimized conditions were 0.19-20.0, 1.11-50.0, 0.02-2.0, 0.15-20.0, 0.80-20.0, 0.35-50.0, and 0.60-50.0 microg/L, respectively. The limits of quantification were, expressed in nanograms per gram of dry weight, 12.6, 74.0, 1.34, 10.0, 53.4, 23.4, and 40.0, respectively. For all of the metals the precision of the instrumental method was <6.3% and that of the analytical method was always <10%, except for aluminum, for which the precision was 12%. The accuracy of the method was evaluated according to the standard additions method, the recoveries being >90% for all of the added concentrations. An interference study was also carried out in a simulated matrix, and it was verified that the deviations of the expected values were <6% for all of the metals. The method was applied to the monitoring of the residues of the referred metals in olive fruits collected from trees pulverized with three different copper formulations available on the market to control fungal diseases. PMID- 16719517 TI - Flavonoid glycosides in bergamot juice (Citrus bergamia Risso). AB - A comprehensive profile of flavonoids in bergamot juice was obtained by a single DAD-ESI-LC-MS-MS course. Eight flavonoids were found for the first time, five of these are C-glucosides (lucenin-2, stellarin-2, isovitexin, scoparin, and orientin 4'-methyl ether), and three are O-glycosides (rhoifolin 4'-O-glucoside, chrysoeriol 7-O-neohesperidoside-4'-O-glucoside, and chrysoeriol 7-O neohesperidoside). A method is proposed to differentiate chrysoeriol and diosmetin derivatives, which are often indistinguishable by LC-MS-MS. In-depth knowledge of the flavonoid content is the starting point for bergamot juice exploitation in food industry applications. PMID- 16719518 TI - Antifungal activities of major tea leaf volatile constituents toward Colletorichum camelliae Massea. AB - A crude glycosidic fraction was prepared from fresh tea leaves and treated with the crude tea enzyme, fractions of cis-3-hexenol, linalool oxide I (cis furanoid), linalool oxide II (trans-furanoid), linalool, methyl salicylate, geraniol, benzyl alcohol, and 2-phenylethanol were monitored to be the major aglycone moieties by analyzing the released volatiles. The amount of the released aglycone moieties is 5.8 times higher than those in free form. For investigation of the functions of the glycosidically bound form aroma constituents in tea leaves, their antifungal activities were determined by antifungal assay. Geraniol, linalool, methyl salicylate, benzyl alcohol, and 2-phenylethanol exhibited significant antifungal activities toward Colletorichum camelliae Massea, although cis-3-hexenol and linalool oxides showed weaker activities by comparison. Among them, geraniol was shown to be the most potential antifungal substance with a MIC value of 440 microg/mL. The crude glycosidic fraction prepared from tea leaves also exhibited significant antifungal activities in a wide range of concentrations from 2 to 25 mg/mL in a PDA medium. It was deduced that the glycosidically bound volatiles are formed and stored in the intact tissue of tea leaf and hydrolyzed by the actions of both the endogenous and the exogenous glycosidases to release volatiles as antifungal substances when exposed to Colletorichum camelliae Massea. The results suggested that the higher content of the bound form geraniol in tea leaves of var. sinensis might be responsible for their stronger antipathogen properties toward tea leaf blight, as opposed to those of var. assamica. PMID- 16719519 TI - Kinetic modeling of 2,4-dichlorophenoxyacetic acid (2,4-D) degradation in soil slurry by anodic fenton treatment. AB - Anodic Fenton treatment (AFT) has been shown to be a promising technology in pesticide wastewater treatment. However, no research has been conducted on the AFT application to contaminated soils. In this study, the 2,4-D degradation kinetics of AFT in a silt loam soil slurry were investigated for the first time, and the effects of various experimental conditions including initial 2,4-D concentration, Fenton reagent delivery rate, amount of humic acid (HA) addition, and pH were examined. The 2,4-D degradation in soil slurry by AFT was found to follow a two-stage kinetic model. During the early stage of AFT (the first 4-5 min), the 2,4-D concentration profile followed a pseudo-first-order kinetic model. In the later stage (typically after 5 or 6 min), the AFT kinetic model provided a better fit. This result is most likely due to the existence of (*)OH scavengers and 2,4-D sorption on soil. The Fe(2+) delivery rate was shown to be a more significant factor in degradation rate than the H(2)O(2) delivery rate when the Fe(2+)/H(2)O(2) ratios were in the range of 1:2 to 1:10. The presence of HA in soil lowered the AFT rate, most probably due to the competition with 2,4-D for consumption of (*)OH and increased sorption of 2,4-D on soil. The optimal pH for 2,4-D degradation in soil slurry by AFT was observed to be in the range of pH 2 3. PMID- 16719520 TI - Studies on water transport through the sweet cherry fruit surface. 10. Evidence for polar pathways across the exocarp. AB - Water uptake through the fruit surface is considered as an important factor in cracking of sweet cherry (Prunus avium L.) fruit. Uptake may occur by diffusion and/or viscous flow along a polar pathway. To establish the mechanism of water uptake, the effects of viscosity and molecular weight of selected osmotica on water uptake into detached sweet cherry fruit were investigated. In addition we investigated the effect of temperature on penetration of 2-(1-naphthyl)[1 (14)C]acetic acid ([(14)C]NAA; pK(a) = 4.2) as a molecular probe in the non dissociated (pH 2.2) and dissociated (pH 6.2) forms. Rates of water uptake were linearly related to the inverse viscosity of gum arabic solutions (range of concentrations and dynamic viscosities 10-300 g L(-1) and 1.3 x 10(-3) to 115.9 x 10(-3) Pa s, respectively). When fruit was incubated in solutions of osmotica of differing molecular weight that were isotonic to the fruit's water potential, water uptake depended on the molecular weight of the osmoticum [range 58-6000 for NaCl to poly(ethylene glycol) 6000 (PEG 6000)]. There was no uptake from PEG 6000 solutions, but rates of water uptake increased as the molecular weight of the osmotica decreased. Apparent water potentials of sweet cherry fruit, determined by incubating fruit in concentration series of selected osmotica, increased as the molecular weight of the osmotica increased up to 1500 and remained constant between 1500 and 6000. Reflection coefficients (sigma) estimated from this relationship were closely related to hydrodynamic radii (r) of the osmotica [sigma = 1.0(+/-0.0) - [10.9(+/-0.9) x 10(-11)][r(-1) (m(-1))], R(2) = 0.97, P < 0.0001]. The permeability of the sweet cherry fruit exocarp to NAA (pK(a) = 4.2) and temperature dependence of NAA permeability (P(d)) as indexed by the energy of activation (E(a), temperature range 5-35 degrees C) were significantly higher for the non-dissociated NAA (pH 2.2, P(d) = 10.2(+/-0.8) x 10(-8) m s(-1), E(a) = 67.0 +/- 1.7 kJ mol(-)(1)) than for the dissociated NAA (pH 6.2, P(d) = 1.1(+/ 0.2) x 10(-8) m s(-1), E(a) = 51.8 +/- 1.9 kJ mol(-)(1)). The activation energy for penetration of the dissociated NAA was closely related to the stomatal density (R( 2) = 0.84, P < 0.0001) but less so for the non-dissociated NAA (R(2) = 0.30, P < 0.03). These data provide evidence for the presence of polar pathways through the sweet cherry fruit exocarp that allow water uptake by viscous flow. These pathways offer a potentially useful target for strategies to reduce water uptake and fruit cracking, provided that a technique is identified that selectively "plugs" these pathways. PMID- 16719521 TI - Anaerobic biotransformation of organo-arsenical pesticides monomethylarsonic acid and dimethylarsinic acid. AB - Monomethylarsonic acid (MMAV) and dimethylarsinic acid (DMAV) are extensively utilized as pesticides, introducing large quantities of arsenic into the environment. Once released into the environment, these organo-arsenicals are subject to microbial reactions. Aerobic biodegradation of MMAV and DMAV has been evaluated, but little is known about their fate in anaerobic environments. The objective of this study was to evaluate the biotransformation of MMAV and DMAV in anaerobic sludge. Biologically mediated conversion occurred under methanogenic or sulfate-reducing conditions but not in the presence of nitrate. Monomethylarsonous acid (MMAIII) was consistently observed as an important metabolite of MMAV degradation, and it was recovered in molar yields ranging from 5 to 47%. The main biotransformation product identified from DMAV metabolism was MMAV, which was recovered in molar yields ranging from 8 to 65%. The metabolites indicate that reduction and demethylation are important steps in the anaerobic bioconversion of MMAV and DMAV, respectively. PMID- 16719523 TI - Selection of potential impact odorants and sensory validation of their importance in typical chardonnay wines. AB - The aim of the present study was to validate the joint sensory impact of target compounds on the typicality degree of wine. Target compounds were selected from previous gas chromatography-olfactometry analysis. The preliminary experiment consisted in selecting odorants thought to have a positive effect on typical Chardonnay wines. Two sets of target compounds were chosen with regard to expected relationships between their concentrations and typicality scores. Target compounds were quantified in 20 wines. The second experiment was dedicated to the sensory evaluation of aroma models obtained by supplementation in wines. Three Chardonnay wines with intermediate typicality scores were supplemented with 6- or 10-compound combinations. The typicality degree of 24 samples was assessed by expert orthonasal perception. Wines supplemented with the 6-compound combinations were judged to be intermediate, whereas wines including the 10-compound combinations were considered to be quite representative of the Chardonnay concept. Such results confirm the active contribution of the 10 combined target compounds to typical Chardonnay wines. PMID- 16719522 TI - Effects of dissolved organic matter on permethrin bioavailability to Daphnia species. AB - Synthetic pyrethroids are widely used insecticides in both agricultural and urban environments. Recent studies show frequent appearances of pyrethroid residues in runoff effluents and sediments, which stimulated concerns over the potential ecotoxicological implications. Pyrethroids are known to have two contrasting characteristics, high aquatic toxicity and strong affinity for the solid phase, that may negate the actual toxicity in a multiphased system. This study evaluated the effect of dissolved organic matter (DOM) on the pyrethroid uptake by and acute toxicity to water-column invertebrates using permethrin as a model compound. During the bioassays, the freely dissolved permethrin concentration was simultaneously measured using poly(dimethylsiloxane) (PDMS) fibers as a biomimetic surrogate. The presence of DOM consistently decreased permethrin uptake and increased its LC(50). For instance, compared to the DOM-free treatment, the LC(50) of permethrin to Ceriodaphnia dubia in a pond water containing DOM at 10 mg L(-)(1) increased from 0.56 to 1.03 microg L(-)(1), whereas the bioaccumulation factor by Daphnia magna decreased by 56%. Permethrin accumulation on the PDMS fiber closely mimicked permethrin uptake by D. magna. Statistical analyses suggest that permethrin associated with DOM was completely unavailable to D. magna or C. dubia. The effect of DOM on permethrin bioavailability appeared to depend also on the source of the DOM. These results indicate that the inhibitory role of DOM should be considered in the development of toxicologically relevant water quality limits and in monitoring protocols for permethrin and other pyrethroids in runoff effluents and surface streams that ubiquitously contain DOM. PMID- 16719524 TI - Sorption behavior of volatile phenols at the oak wood/wine interface in a model system. AB - The sorption in a model system of aroma compounds of enological interest (mixture of the eight derivatives from guaiacol, 4-ethylphenol, and whiskylactone) onto wood was investigated to assess the influence of wood on the concentration of these volatiles during the aging of wine. To evaluate the influence of the solubility of aroma compounds in sorption phenomena, this parameter was determined for each volatile compound in model wine at 10 and 25 degrees C. The solubility is significantly higher in the model wine than in water and remains constant in the range of temperatures studied, except for guaiacol and vanillin. Kinetic and equilibrium sorptions were investigated. Sorption kinetics showed that the sorption equilibrium for all aroma compounds was reached after 6-7 days. From the study of the individual sorption isotherms, two distinct kinds of sorption behavior were observed depending on the presence or not of an ethylenic para substituent conjugated to the phenyl ring. K(ww) partition coefficients between the wood and the model wine were determined, which exhibited an unusual positive variation with temperature. PMID- 16719525 TI - Comparison of aroma volatiles in commercial Merlot and Cabernet Sauvignon wines using gas chromatography-olfactometry and gas chromatography-mass spectrometry. AB - Seventy-four aroma active compounds were observed in Merlot and Cabernet Sauvignon wines produced in California and Australia. Volatiles were sampled using solid phase microextraction and analyzed using time-intensity gas chromatography-olfactometry and gas chromatography-mass spectrometry (GC-MS). The most intense odorants were 3-methyl-1-butanol, 3-hydroxy-2-butanone, octanal, ethyl hexanoate, ethyl 2-methylbutanoate, beta-damascenone, 2-methoxyphenol, 4 ethenyl-2-methoxy-phenol, ethyl 3-methylbutanoate, acetic acid, and 2 phenylethanol. Aroma compounds were classified according to their aroma descriptor similarity and summed into nine distinct categories consisting of fruity, sulfury, caramel/cooked, spicy/peppery, floral, earthy, pungent/chemical, woody, and green/vegetative/fatty. Both Merlot and Cabernet Sauvignon wines were characterized by high fruity, caramel, green, and earthy aroma totals. Although there were distinct quantitative differences between Merlot and Cabernet wines, the relative aroma category profiles of the four wines were similar. Of the 66 volatiles identified by GC-MS, 28 were esters and 19 were minor alcohols. Between 81 and 88% of the total MS total ion chromatogram peak areas from each wine type were produced from only eight compounds: ethanol, ethyl octanoate, ethyl decanoate, ethyl acetate, 3-methyl-1-butanol, ethyl hexanoate, diethyl succinate, and 2-phenylethanol. Merlot wines from both Australia and California contained 4 5 times more ethyl octanoate than Cabernet Sauvignon wines from the same sources. PMID- 16719526 TI - Complex viscosity induced by protein composition variation influences the aroma release of flavored stirred yogurt. AB - Dairy protein composition is known to influence the structure and the texture characteristics of yogurt. The objective of the present work was therefore to investigate the impact of protein composition, at a constant protein level, on the physicochemical properties of 4% fat flavored stirred yogurt and, more specifically, on the rheological properties, the microstructure, and the aroma release. The results showed that caseinate-enriched yogurt generally presented changes in their microstructure network and had a higher complex viscosity than whey protein-enriched yogurt. To a lesser extent, the release of the majority of aroma compounds was lower in caseinate-enriched yogurt. It was therefore possible to quantify physicochemical interactions between aroma compounds and proteins. The influence of gel structure on the flavor release was observed and was in agreement with sensory characteristics previously studied for these products. PMID- 16719527 TI - Ontogenic profiling of glucosinolates, flavonoids, and other secondary metabolites in Eruca sativa (salad rocket), Diplotaxis erucoides (wall rocket), Diplotaxis tenuifolia (wild rocket), and Bunias orientalis (Turkish rocket). AB - As an influence of the Mediterranean diet, rocket species such as Eruca sativa L., Diplotaxis species, and Bunias orientalis L. are eaten all over the world at different ontogenic stages in salads and soups. They are all species within the plant order Capparales (glucosinolate-containing species), and all are from the family Brassicaceae. Predominantly, the leaves of these species are eaten raw or cooked, although Eruca flowers are also consumed. There is considerable potential with raw plant material for a higher exposure to bioactive phytochemicals such as glucosinolates, their hydrolysis products, and also phenolics, flavonoids, and vitamins such as vitamin C. These compounds are susceptible to ontogenic variation, and the few published studies that have addressed this topic have been inconsistent. Thus, an ontogenic study was performed and all samples were analyzed using a previously developed robust liquid chromatography/mass spectrometry method for the identification and quantification of the major phytochemicals in all tissues of the rocket species. Seeds and roots of both Eruca and Diplotaxis contained predominantly 4-methylthiobutylglucosinolate. Leaves of Eruca and Diplotaxis contained high amounts of 4 mercaptobutylglucosinolate with lower levels of 4-methylthiobutlyglucosinolate and 4-methylsulfinylbutylglucosinolate. Flowers of Eruca and Diplotaxiscontained predominantly 4-methylsulfinylbutyl-glucosinolate. In addition, roots of both Diplotaxisspecies contained 4-hydroxybenzylglucosinolate but 4 hydroxybenzylglucosinolate was absent from roots of Eruca. Seeds and seedlings of all Eruca contained N-heterocyclic compounds but no sinapine, whereas Diplotaxis contained sinapine but not the N-heterocycles. In all tissues of B. orientalis, 4 hydroxybenzylglucosinolate and 4-methylsulfinyl-3-butenylglucosinolate were predominant. All rocket tissues, except roots, contained significant levels of polyglycosylated flavonoids, with/without hydroxycinnamoyl acylation. The core aglycones were kaempferol, quercetin, and isorhamnetin. The exception was B. orientalis, which had a negligible seed flavonoid content as compared with the other species. Anthocyanins were only detected in Eruca flowers and consisted of a complex pattern of at least 16 different anthocyanins. PMID- 16719528 TI - Extraction of polyphenols from processed black currant (Ribes nigrum L.) residues. AB - The total phenol and anthocyanin contents of black currant pomace and black currant press residue (BPR) extracts, extracted with formic acid in methanol or with methanol/water/acetic acid, were studied. Anthocyanins and other phenols were identified by means of reversed phase HPLC, and differences between the two plant materials were monitored. In all BPR extracts, phenol levels, determined by the Folin-Ciocalteu method, were 8-9 times higher than in the pomace extracts. Acid hydrolysis liberated a much higher concentration of phenols from the pomace than from the black currant press residue. HPLC analysis revealed that delphinidin-3-O-glucoside, delphinidin-3-O-rutinoside, cyanidin-3-O-glucoside, and cyanidin-3-O-rutinoside were the major anthocyanins and constituted the main phenol class ( approximately 90%) in both types of black currant tissues tested. However, anthocyanins were present in considerably lower amounts in the pomace than in the BPR. In accordance with the total phenol content, the antioxidant activity determined by scavenging of 2,2'-azinobis(3-ethylbenzothiazoline-6 sulfonic acid) radical cation, the ABTS(*)(+) assay, showed that BPR extracts prepared by solvent extraction exhibited significantly higher (7-10 times) radical scavenging activity than the pomace extracts, and BPR anthocyanins contributed significantly (74 and 77%) to the observed high radical scavenging capacity of the corresponding extracts. PMID- 16719529 TI - Dose-dependent influence of commercial garlic (Allium sativum) on rats fed cholesterol-containing diet. AB - The aim of this investigation was to evaluate the dose-dependent influence of commercial garlic on rats fed cholesterol-containing diets. It was found that commercial garlic contains high concentrations of dietary fibers, microelements, and total polyphenols, and its total antioxidant capacity as determined by two independent assays [1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azinobis(3 ethylbenzthiazoline-6-sulfonic acid) (ABTS)] was similar to that of the original garlic samples. Wistar rats (35) were randomly divided into five diet groups, named control, Chol, Garlic500, Garlic750, and Garlic1000. Control rats were fed basal diet (BD), which included wheat starch, casein, soybean oil, and vitamin and mineral mixtures. To the BD of the Chol group was added 1% of cholesterol. To the BD of the other three groups (Garlic500, Garlic750, and Garlic1000) were added 1% of cholesterol and commercial garlic equal to 500, 750, and 1000 mg of raw garlic per kilogram of animal weight. After 4 weeks of the experiment only in rats from the Garlic500 group were a significant hindering in the rise in plasma lipids and also a significant hindering in a decrease of plasma antioxidant activity registered. A significant decrease in plasma circulating fibrinogen and an increase in the clotting time were found in the same group of rats (P < 0.05 in both cases). The fibrinogenolytic effect of garlic diets was visualized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In the fibrinogen fraction of Garlic500 the 66, 24, and 14 kDa protein bands were detected with weaker protein intensity than in the corresponding ones in the Garlic750 and Garlic1000 diet groups. In conclusion, the positive influences of commercial garlic on plasma lipids, proteins, antioxidant activity, and some indices of blood coagulation are dose-dependent. Therefore, commercial garlic (Elena, Zelazkow, Poland) could be a valuable component of atherosclerosis-preventing diets only in optimal doses. PMID- 16719530 TI - Wheat-kernel-associated endoxylanases consist of a majority of microbial and a minority of wheat endogenous endoxylanases. AB - The endoxylanases associated with wheat kernels consist of wheat endogenous endoxylanases on one hand and kernel-associated microbial endoxylanases on the other hand. Assessment of their presence, based on analysis of their enzymic activity, can be expected to be hampered by the presence in wheat of high levels of endogenous endoxylanase inhibitors, which are able to inhibit the wheat-kernel associated microbial endoxylanases. On the basis of preliminary experiments aimed at clarifying the distribution of the wheat-associated endoxylanases, a method to estimate total endoxylanase activities in wheat kernels was developed. Extensive washing of wheat kernels with universal buffer of pH 8.0 provided near quantitative separation of the microbial endoxylanases located on the surface of wheat kernels from the endogenous endoxylanases and endoxylanase inhibitors located in such kernels. The microbial or endogenous nature of the endoxylanases was confirmed by making use of the inhibition specificity of endoxylanase inhibitors. Determination of the endoxylanase activity in the washing liquid, corresponding to the microbial endoxylanase population, and the washed kernels, corresponding to the endogenous endoxylanase population, allowed estimation of the total endoxylanase activities associated with the wheat kernel. Results showed that microbial endoxylanases can account for over 90% of the total wheat associated endoxylanase activity and that the latter can be at least 5 times higher than the apparent endoxylanase activity. PMID- 16719531 TI - Chlorophyll catabolism pathway in fruits of Capsicum annuum (L.): stay-green versus red fruits. AB - The aim of the present study is to investigate the chlorophyll catabolism pathway of wild-type red and stay-green mutants of Capiscum annuum (L.) fruits. In the wild-type red lines chlorophyll catabolism is concomitant with the start of carotenogenesis, whereas in the stay-green mutant lines the chlorophylls coexist with that process, even in over-ripe fruit. During the first stages of ripening, the chlorophyll a/chlorophyll b ratio is similar for both genotypes, but as ripening proceeds, the ratio in the stay-green lines becomes very high as a result of a blocked degradation of chlorophyll a while chlorophyll b is degraded at a normal rate. The absence of dephytylated chlorophylls in the wild-type lines distinguishes these from the mutant lines, in which there is a sequential accumulation of chlorophyllide a and pheophorbide a. Allomerized chlorophylls (13(2)-OH-chlorophyll a and b) have also been identified in the catabolic process of the mutant lines, but are absent from the wild type. Consequently, an alteration in pheophorbide a oxygenase (PaO) activity seems to be responsible for the stay-green genotype in the lines of pepper analyzed in this study. PMID- 16719532 TI - Effect of cyanidin-3-O-glucoside on UVB-induced response in human keratinocytes. AB - One of the most significant risk factors associated with the development of skin disease is exposure to UVB radiation from the sun. In particular, UVB light can activate inflammatory and apoptotic pathways, leading to skin damage. Anthocyanins, a group of flavonoids present in many common vegetable foods, are known for their chemopreventive activity. The aim of this study was to evaluate the efficacy of cyanidin-3-O-glucoside (C3G) on modulation of cellular responses following exposure to UVB doses in human keratinocytes (HaCaT). In our study, UVB exposed cells showed significant increase of the translocation of transcription factors NF-kB and AP-1, overexpression of the proinflammatory cytokine IL-8, cleavage of procaspase-3 (a key step in apoptotic pathway), and DNA fragmentation. All these effects elicited by UVB exposure were clearly inhibited by pretreating HaCaT cells with C3G. In conclusion, our data demonstrate that C3G can protect skin cells against the adverse effects of UVB radiation and suggest that it might successfully be employed as a skin photoprotective agent. PMID- 16719533 TI - Structural identification and distribution of proanthocyanidins in 13 different hops. AB - Ten newly isolated hop proanthocyanidin oligomers and flavan-3-ol monomers from 13 different hops have been identified as gallocatechin, gallocatechin-(4alpha- >8)-catechin, gallocatechin-(4alpha-->6)-catechin, catechin-(4alpha-->8) gallocatechin, catechin-(4alpha-->6)-gallocatechin, afzelechin-(4alpha-->8) catechin, catechin-(4alpha-->8)-catechin-(4alpha-->8)-catechin, epicatechin (4beta-->8)-epicatechin-(4beta-->8)-catechin, catechin-(4alpha-->8)-gallocatechin (4alpha-->8)-catechin, and gallocatechin-(4alpha-->8)-gallocatechin-(4alpha-->8) catechin, together with seven previously isolated oligomers, namely, catechin, epicatechin, epicatechin-(4beta-->8)-catechin, epicatechin-(4beta-->8) epicatechin, catechin-(4alpha-->8)-catechin, catechin-(4alpha-->8)-epicatechin, and epicatechin-(4beta-->8)-catechin-(4alpha-->8)-catechin. These compounds were subjected to acid-catalyzed degradation in the presence of phloroglucinol or by partial or complete acid-catalyzed degradation and reaction with benzyl mercaptan followed by desulfurization. The resultant adducts when compared to authentic samples by high-performance liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometry and high-performance liquid chromatography electrospray ionization tandem mass spectrometry served to identify the precursors. The composition of proanthocyanidins from 13 different hops was similar, but the concentration of individual compounds showed some differences, which indicated that hop proanthocyanidin profiles are affected by geographic origin and are variable depending on the cultivars. PMID- 16719534 TI - Procyanidin and catechin contents and antioxidant capacity of cocoa and chocolate products. AB - Cocoa and chocolate products from major brands were analyzed blind for total antioxidant capacity (AOC) (lipophilic and hydrophilic ORAC(FL)), catechins, and procyanidins (monomer through polymers). Accuracy of analyses was ascertained by comparing analyses on a NIST standard reference chocolate with NIST certified values. Procyanidin (PC) content was related to the nonfat cocoa solid (NFCS) content. The natural cocoa powders (average 87% of NFCS) contained the highest levels of AOC (826 +/- 103 micromol of TE/g) and PCs (40.8 +/- 8.3 mg/g). Alkalized cocoa (Dutched powders, average 80% NFCS) contained lower AOC (402 +/- 6 micromol of TE /g) and PCs (8.9 +/- 2.7 mg/g). Unsweetened chocolates or chocolate liquor (50% NFCS) contained 496 +/- 40 micromol of TE /g of AOC and 22.3 +/- 2.9 mg/g of PCs. Milk chocolates, which contain the least amount of NFCS (7.1%), had the lowest concentrations of AOC (80 +/- 10 micromol of TE /g) and PCs (2.7 +/- 0.5 mg/g). One serving of cocoa (5 g) or chocolate (15 or 40 g, depending upon the type of chocolate) provides 2000-9100 micromol of TE of AOC and 45-517 mg of PCs, amounts that exceed the amount in a serving of the majority of foods consumed in America. The monomers through trimers, which are thought to be directly bioavailable, contributed 30% of the total PCs in chocolates. Hydrophilic antioxidant capacity contributed >90% of AOC in all products. The correlation coefficient between AOC and PCs in chocolates was 0.92, suggesting that PCs are the dominant antioxidants in cocoa and chocolates. These results indicate that NFCS is correlated with AOC and PC in cocoa and chocolate products. Alkalizing dramatically decreased both the procyanidin content and antioxidant capacity, although not to the same extent. PMID- 16719535 TI - Antioxidant activity and polyphenol and procyanidin contents of selected commercially available cocoa-containing and chocolate products in the United States. AB - In the United States, commercially available foods, including cocoa and chocolate, are being marketed with statements referring to the level of antioxidant activity and polyphenols. For cocoa-containing foods, there has been no comprehensive survey of the content of these and other chemistries. A survey of cocoa and chocolate-containing products marketed in the United States was conducted to determine antioxidant activity and polyphenol and procyanidin contents. Commercially available samples consisted of the top market share products in each of the following six categories: natural cocoa, unsweetened baking chocolate, dark chocolate, semisweet baking chips, milk chocolate, and chocolate syrup. Composite samples were characterized using four different methods: oxygen radical absorbance capacity (ORAC), vitamin C equivalence antioxidant capacity (VCEAC), total polyphenols, and procyanidins. All composite lots were further characterized for percent nonfat cocoa solids (NFCS) and percent fat. Natural cocoas had the highest levels of antioxidant activities, total polyphenols, and procyanidins followed by baking chocolates, dark chocolates and baking chips, and finally milk chocolate and syrups. The results showed a strong linear correlation between NFCS and ORAC (R (2) = 0.9849), total polyphenols (R (2) = 0.9793), and procyanidins (R (2) = 0.946), respectively. On the basis of principal component analysis, 81.4% of the sample set was associated with NFCS, antioxidant activity, total polyphenols, and procyanidins. The results indicated that, regardless of the product category, NFCS were the primary factor contributing to the level of cocoa antioxidants in the products tested. Results further suggested that differences in cocoa bean blends and processing, with the possible exception of Dutching, are minor factors in determining the level of antioxidants in commercially available cocoa-containing products in the United States. PMID- 16719536 TI - Concentrations of anthocyanins in common foods in the United States and estimation of normal consumption. AB - Anthocyanins (ACNs) are water-soluble plant pigments that have important functions in plant physiology as well as possible health effects. Over 100 common foods were screened for ACNs, and 24 of them were found to contain ACNs. Concentrations of total ACNs varied considerably from 0.7 to 1480 mg/100 g of fresh weight in gooseberry ('Careless' variety) and chokeberry, respectively. Not only does the concentration vary, but the specific anthocyanins present in foods are also quite different. Only six common aglycones, delphinidin, cyanidin, petunidin, pelargonidin, peonidin, and malvidin, were found in all of these foods. However, their sugar moieties and acylation patterns varied from food to food. Results from this study will add to the available data for the USDA Nutrient Database of flavonoids. On the basis of the concentration data and updated food intake data from NHANES 2001-2002, the daily intake of ACNs is estimated to be 12.5 mg/day/person in the United States. Of the different aglycones, cyanidin, delphinidin, and malvidin were estimated to contribute 45, 21, and 15%, respectively, of the total ACN intake. Nonacylated contributed 77% compared to 23% from acylated ACNs. PMID- 16719539 TI - Transdermal scopolamine for prevention of motion sickness : clinical pharmacokinetics and therapeutic applications. AB - A transdermal therapeutic system for scopolamine (TTS-S) was developed to counter the adverse effects and short duration of action that has restricted the usefulness of scopolamine when administered orally or parenterally. The plaster contains a reservoir of 1.5 mg of scopolamine programmed to deliver 0.5 mg over a 3-day period. A priming dose (140 microg) is incorporated into the adhesive layer to saturate certain binding sites within the skin and to accelerate the achievement of steady-state blood levels. The remainder is released at a constant rate of approximately 5 microg/hour. The protective plasma concentration of scopolamine is estimated to be 50 pg/mL. TTS-S attains that concentration after 6 hours; a steady state of about 100 pg/mL is achieved 8-12 hours after application. Yet 20-30% of subjects failed to attain the estimated protective concentration, and plasma concentrations measured in subjects who failed to respond to TTS-S were lower than in responders. These findings may explain some of the treatment failures. Overall, the product appears to be the approximate functional equivalent of a 72-hour slow intravenous infusion. A combination of transdermal and oral scopolamine (0.3 or 0.6 mg) was effective and well tolerated in producing desired plasma concentrations 1-hour post-treatment. TTS-S has proved to be significantly superior to placebo in reducing the incidence and severity of motion sickness by 60-80%. It was more effective than oral meclizine or cinnarizine, similar to oral scopolamine 0.6 mg or promethazine plus ephedrine, and the same as or superior to dimenhydrinate. The addition of ephedrine or the use of two patches did not improve its efficacy, but rather increased the rate of adverse effects. TTS-S was most effective against motion sickness 8-12 hours after application. Despite previous evidence to the contrary, a recent bioavailability study demonstrated similar intraindividual absorption and sustained clinical efficacy with long-term use of the drug. The adverse effects produced by TTS-S, although less frequent, are qualitatively typical of those reported for the oral and parenteral formulations of this agent. Dry mouth occurs in about 50-60% of subjects, drowsiness in up to 20%, and allergic contact dermatitis in 10%. Transient impairment of ocular accommodation has also been observed, in some cases possibly the result of finger-to-eye contamination. Low dose pyridostigmine was found effective in preventing cycloplegia but not mydriasis. Adverse CNS effects, including toxic psychosis (mainly in elderly and paediatric patients), have been reported only occasionally, as have difficulty in urinating, headache, rashes and erythema. Adverse effects were not correlated with plasma scopolamine concentrations. TTS-S produced only about half the incidence of drowsiness caused by oral dimenhydrinate or cinnarizine, and a level of adverse effects similar to that found with oral meclizine. Performance is not affected by short-term use. Prolonged or repeated application may cause some impairment of memory storage for new information. However, sea studies revealed significantly less reports of a decrement in performance or drowsiness due to prevention of sea sickness. The recommended dosage is a single TTS-S patch applied to the postauricular area at least 6-8 hours before the anti-motion sickness effect is required. For faster protection, the patch may be applied 1 hour before the journey in combination with oral scopolamine (0.3 or 0.6 mg). After 72 hours, the patch should be removed and a new one applied behind the opposite ear. Its place in therapy is mainly on long journeys (6-12 hours or longer), to avoid repeated oral doses, or when oral therapy is ineffective or intolerable. PMID- 16719540 TI - How may anticancer chemotherapy with fluorouracil be individualised? AB - Fluorouracil is used clinically against various solid tumours. Both fluorouracil toxicity and pharmacokinetics vary highly within and between individuals. The reasons why doses are not individualised routinely are difficulties in defining, predicting and achieving an optimal fluorouracil exposure or dose because of a narrow therapeutic index, nonlinear pharmacokinetics, variabilities in administration rates and metabolism, and in targets like thymidylate synthase. To individualise fluorouracil administration before the first dose, assessment of the individual dihydropyrimidine dehydrogenase (DPD) activity may be useful, because this genetically highly polymorphic enzyme controls approximately 80% of fluorouracil elimination. A complete or partial loss of DPD activity in 0.1 and 3 5% of Caucasians, respectively, leads to increased fluorouracil exposure and toxicity. Several methods to assess DPD activity in patients have been proposed (genotyping, various phenotyping methods), but each of them has limitations, as has the fluorouracil test dose approach. To adapt exposure towards fluorouracil a priori, a combination of genotyping and phenotyping may yield better prediction of toxicity than one method alone. A prerequisite for dose adaptation is the definition of fluorouracil exposure ranges with sufficient therapeutic activity, but without serious toxicity. While an increased risk of leukopenia, diarrhoea, stomatitis, and hand-foot syndrome during continuous 5-day infusions was related to fluorouracil exposures above an area under the plasma concentration-time curve (AUC) threshold of 25-30 mg.h/L, tumour response was higher when an AUC of approximately 30 mg.h/L was achieved, illustrating the extremely narrow therapeutic window of fluorouracil. Pharmacokinetic target values are less clear for other regimens, including chronomodulated regimens, which yielded a superior clinically efficacy and tolerability in several trials. However, the monitoring of fluorouracil plasma concentrations seems principally useful for individual a posteriori dose adjustment. Whether an adaptation of the fluorouracil starting dose to the results of two DPD activity tests before fluorouracil administration a priori, and the adaptation of doses to fluorouracil exposure a posteriori is a reasonable approach to better prevent toxicity and increase efficacy, remains to be evaluated in randomised clinical studies comparing these strategies to routine clinical safety monitoring. PMID- 16719541 TI - Possible interethnic differences in omeprazole pharmacokinetics : comparison of Jordanian Arabs with other populations. AB - BACKGROUND AND OBJECTIVE: Considerable ethnic differences have been reported in the incidence of the poor metaboliser (PM) genotype of cytochrome P450 (CYP) 2C19. The frequency of this genotype was found to be much higher in Oriental persons (13-23%) than in American or European populations (3-5%). There are, however, no valid data published for Arabic subjects. The present study was conducted to evaluate pharmacokinetic parameters of omeprazole after a single dose in healthy Jordanian Arabic subjects and to compare the results with data published for other populations. METHODS: Seventy-four healthy male Jordanian Arabic volunteers contributed to the study, which was performed at Al Essra Hospital in Amman, Jordan. After an overnight fast, omeprazole was administered as a single Losec 20mg capsule. A total of 20 blood samples were collected over a 10-hour period after administration. Omeprazole pharmacokinetic parameters were determined from the plasma concentration-time profiles using the WinNonlin software. Kolmogorov-Smirnov's test and probit plots of omeprazole area under the plasma concentration-time curve (AUC) data were used to analyse the frequency distribution of phenotypic data. RESULTS: The mean pharmacokinetic parameters and their corresponding coefficient of variation (CV%) for peak plasma concentration (Cmax), AUC from time zero to infinity (AUCinfinity), time to reach Cmax (tmax), apparent oral clearance (CL/F) and elimination half-life (t(1/2)) were 314.96 ng/mL (56%), 923.2 ng . h/mL (108.6%), 2.1h (44%), 0.66 L/h/kg (92%) and 1.5 h (56.6%), respectively. Interindividual differences in the current study were high for all pharmacokinetic parameters, yet comparable to CVs reported in nonphenotyped subjects identified within other ethnic groups (40.3-159% for AUC and 39-48.2% for Cmax). The frequency distribution of all parameters, particularly the AUC, was shown to be trimodal. This has proposed the presence of three distinct phenotypes, designated as extensive metabolisers (EMs), slow extensive metabolisers (SEMs), and PMs, with corresponding frequency of 36.5%, 39.2% and 24.3%, respectively. After stratification, the relative mean AUCs of omeprazole in EMs, SEMs and PMs were 1 : 2.7 : 9.3 (all p < 0.001). Accordingly, the CL/F of omeprazole showed a ratio of 9.8 : 3.6 : 1 for three phenotype groups, respectively. For other pharmacokinetic parameters including Cmax, t1/2, AUC normalised for bodyweight (AUCN), Cmax/dose and AUC/dose, there were also significant differences between the three groups. CONCLUSIONS: The current pharmacokinetic study revealed that the majority of the Jordanian Arabics seemed to be more properly classified within the EM phenotype. More specifically, the observed metabolic rates of heterozygous and homozygous Jordanian Arabic EMs were more comparable to those of Caucasian EMs than Oriental EMs. Consequently, higher dosage requirements can be expected among most of the Jordanian Arabics. Yet, the incidence of PMs is significant and they seemed to exhibit a similar pharmacokinetic pattern to Chinese PMs in terms of long-term exposure (clearance and AUC) as well as short-term exposure (Cmax) parameters, after adjustment for dose and bodyweight. Therefore, further clinical application of CYP2C19 polymorphism is anticipated in Jordanian Arabic mixed population, particularly if long-term use of omeprazole is intended. PMID- 16719542 TI - Predicting the toxicity of weekly docetaxel in advanced cancer. AB - OBJECTIVES: To determine the safety profile of 40 mg/m(2) docetaxel administered weekly to a mixed population of advanced cancer patients and identify predictors of toxicity and survival following treatment with weekly docetaxel in this population. PATIENTS AND METHODS: 68 patients with advanced cancer were enrolled into the study. Various patient characteristics, including inflammatory and nutritional status, docetaxel pharmacokinetics and liver function were investigated. Predictors of treatment-related toxicity and survival were analysed using multivariate logistic regression and Cox proportional hazards analysis, respectively. RESULTS: 27 patients (40%) experienced grade 3 or 4 toxicity, mainly gastrointestinal toxicities (20%), leukopenia (16%) and neutropenia (12%), during the first 8 weeks of docetaxel treatment. Docetaxel pharmacokinetics were the only predictive factor for haematological toxicity. The odds of severe haematological toxicity were approximately 9-fold higher for patients with reduced docetaxel clearance (e.g. <30 L/h). The odds of non-haematological toxicity were about 3-fold higher for patients with elevated levels of inflammatory markers: alpha(1)-acid glycoprotein (AAGP) >1.5 g/L or C-reactive protein >10 mg/L). Multivariate analysis indicated that weight loss, liver dysfunction and elevated levels of AAGP were independently significant predictors of survival. CONCLUSION: This is the first description of factors predictive of the toxicity and survival following weekly administration of docetaxel. Patients with reduced clearance of docetaxel and elevated markers of inflammation experienced worse adverse effects, while patients with weight loss, liver dysfunction and elevated markers of inflammation had worse survival. PMID- 16719543 TI - Cost-efficient higher-order crossover designs in comparative bioavailability studies. AB - BACKGROUND AND OBJECTIVE: Cost is an extremely important factor to consider when planning drug clinical trials. Higher-order crossover designs have recently drawn considerable attention in comparative bioavailability studies because of their desirable statistical properties. In this paper, we compared the cost efficiency of five commonly used higher-order crossover designs under certain cost function for comparative bioavailability studies. METHODS: Multivariate normal data were simulated under scenarios of a wide range of variability and correlations (coefficient of variation = 10-40%; correlation coefficient rho = 0.2-0.8). Monte Carlo simulations and mixed-effects models were carried out to obtain empirical sample sizes for each design using Schuirmann's two-one sided test procedure, under an 80% power and a 5% significance level, based on the US FDA bioequivalence criteria (80-125%). The five crossover designs studied were the two-period four-sequence (D2 x 4), the three-period two-sequence (D3 x 2), the three-period four-sequence (D3 x 4), the four-period two-sequence (D4 x 2), and the four-period four-sequence (D4 x 4). Costs for each design were then determined by a cost function, which takes into account costs for recruiting and screening, costs associated with period, and the overheads incurred for multiple sequences. Comparison of the costs for the above-mentioned designs was made under different scenarios. RESULTS: There was no single design uniformly dominating the others in terms of cost efficiency for comparative bioavailability studies. The designs D3 x 2 and D4 x 4 (especially the former) have the best overall performance in terms of cost efficiency for comparative bioavailability studies. They dominated the other designs under most of the scenarios. The design D2 x 4 showed the worst performance among the five crossover designs. CONCLUSIONS: A D3 x 2, and D4 x 2 crossover designs are recommended to achieve cost efficiency with a given power. The D2 x 4 crossover design is not recommended in general for comparative bioavailability studies. PMID- 16719544 TI - Exploring the relationship between expression of cytochrome P450 enzymes and gefitinib pharmacokinetics. AB - BACKGROUND AND OBJECTIVES: Exposure to gefitinib (IRESSA, ZD1839), an epidermal growth factor receptor-tyrosine kinase inhibitor, is highly variable between subjects. In an attempt to explain this variability, three pharmacokinetic studies were carried out in healthy volunteers to investigate the relationship between exposure to gefitinib and cytochrome P450 (CYP) 3A phenotype (study 1), CYP3A5 genotype (study 2) and CYP2D6 genotype (study 3). METHODS: In study 1 all 15 healthy volunteers received single oral doses of midazolam (7.5 mg), as a CYP3A probe, and gefitinib (500 mg), separated by an appropriate washout period. Plasma concentrations of midazolam and gefitinib were measured. In study 2, 73 healthy volunteers with previously defined single-dose gefitinib pharmacokinetic profiles were genotyped for CYP3A5. In study 3 a single oral dose of gefitinib (250 mg) was administered to poor and extensive CYP2D6 metabolisers (n = 15 in each group). Plasma concentrations of gefitinib and its major metabolite, M523595, were measured. Plasma concentrations of gefitinib, M523595 and midazolam were measured using high-performance liquid chromatography with tandem mass spectrometric detection, and appropriate pharmacokinetic parameters were determined by non-compartmental methods. Genetic analysis of CYP3A5 (study 2) and CYP2D6 (study 3) alleles was carried out using standard methodology. RESULTS: In study 1 there was some indication of a correlation between the area under the plasma concentration-time curve from time zero to infinity (AUCinfinity) values of midazolam and gefitinib, although this did not reach statistical significance (p = 0.062, regression analysis). In study 2 eight of 73 volunteers (11%) were identified as CYP3A5 expressers. No apparent relationship was observed between the occurrence of the CYP3A5 expresser genotype and gefitinib plasma clearance or terminal elimination halflife. In study 3 M523595 was not detected in any plasma samples collected from poor CYP2D6 metabolisers. Gefitinib geometric mean AUCinfinity and peak plasma drug concentration were higher in poor CYP2D6 metabolisers compared with extensive metabolisers (AUCinfinity 3060 vs 1430 ng . h/mL, p < 0.05, ANOVA), although the range of values was wide with considerable overlap between the groups. Gefitinib was well tolerated in both groups. CONCLUSIONS: Individual differences in CYP3A expression do not explain all the interindividual variability in gefitinib exposure. There is no apparent relationship between CYP3A5 genotype and gefitinib clearance. The lack of measurable levels of M523595 in poor CYP2D6 metabolisers confirms that production of this metabolite is mediated by CYP2D6. Although higher exposure to gefitinib occurs in individuals who are poor CYP2D6 metabolisers, genotyping prior to initiation of therapy and dosage adjustment are not warranted. PMID- 16719545 TI - NF-kappaB in photodynamic therapy: discrepancies of a master regulator. AB - Tumor eradication by photodynamic therapy (PDT) results from the onset of distinct killing processes. In addition to the well-known necrotic and apoptotic mechanisms, PDT initiates an inflammatory response that will indirectly contribute to tumor clearance. The NF-kappaB transcription factor is a major regulator of inflammation modulating the expression of cytokines, chemokines, and adhesion molecules in various cell types in response to a large number of stimuli. Besides, NF-kappaB regulates the expression of antiapoptotic genes, cyclooxygenases (COXs) and metalloproteinases (MMPs) as well, thereby favoring tumor cell proliferation and dissemination. In the present review, we aim to summarize the current knowledge on NF-kappaB status following photosensitization of cancer cells and endothelial cells. In order to unravel the NF-kappaB impact in PDT tumorigenicity and recurrences, we will stress the discrepancies of this major transcription factor relative to the signaling cascades underlying its activation and the cellular effects triggered by its translocation into the nucleus and its binding to its target genes. PMID- 16719546 TI - Statins in patients with chronic kidney disease: evidence from systematic reviews and randomized clinical trials. PMID- 16719547 TI - Deployment of early diagnosis and mefloquine-artesunate treatment of falciparum malaria in Thailand: the Tak Malaria Initiative. AB - BACKGROUND: Early diagnosis and treatment with artesunate-mefloquine combination therapy (MAS) have reduced the transmission of falciparum malaria dramatically and halted the progression of mefloquine resistance in camps for displaced persons along the Thai-Burmese border, an area of low and seasonal transmission of multidrug-resistant Plasmodium falciparum. We extended the same combination drug strategy to all other communities (estimated population 450,000) living in five border districts of Tak province in northwestern Thailand. METHODS AND FINDINGS: Existing health structures were reinforced. Village volunteers were trained to use rapid diagnostic tests and to treat positive cases with MAS. Cases of malaria, hospitalizations, and malaria-related deaths were recorded in the 6 y before, during, and after the Tak Malaria Initiative (TMI) intervention. Cross sectional surveys were conducted before and during the TMI period. P. falciparum malaria cases fell by 34% (95% confidence interval [CI], 33.5-34.4) and hospitalisations for falciparum malaria fell by 39% (95% CI, 37.0-39.9) during the TMI period, while hospitalisations for P. vivax malaria remained constant. There were 32 deaths attributed to malaria during, and 22 after the TMI, a 51.5% (95% CI, 39.0-63.9) reduction compared to the average of the previous 3 y. Cross sectional surveys indicated that P. vivax had become the predominant species in Thai villages, but not in populations living on the Myanmar side of the border. In the displaced persons population, where the original deployment took place 7 y before the TMI, the transmission of P. falciparum continued to be suppressed, the incidence of falciparum malaria remained low, and the in vivo efficacy of the 3-d MAS remained high. CONCLUSIONS: In the remote malarious north western border area of Thailand, the early detection of malaria by trained village volunteers, using rapid diagnostic tests and treatment with mefloquine-artesunate was feasible and reduced the morbidity and mortality of multidrug-resistant P. falciparum. PMID- 16719549 TI - Mistrust among minorities and the trustworthiness of medicine. PMID- 16719551 TI - Should language matter less to journals? PMID- 16719548 TI - Does random treatment assignment cause harm to research participants? AB - BACKGROUND: Some argue that by precluding individualized treatment, randomized clinical trials (RCTs) provide substandard medical care, while others claim that participation in clinical research is associated with improved patient outcomes. However, there are few data to assess the impact of random treatment assignment on RCT participants. We therefore performed a systematic review to quantify the differences in health outcomes between randomized trial participants and eligible non-participants. METHODS AND FINDINGS: Studies were identified by searching Medline, the Web of Science citation database, and manuscript references. Studies were eligible if they documented baseline characteristics and clinical outcomes of RCT participants and eligible non-participants, and allowed non-participants access to the same interventions available to trial participants. Primary study outcomes according to patient group (randomized trial participants versus eligible non-participants) were extracted from all eligible manuscripts. For 22 of the 25 studies (88%) meeting eligibility criteria, there were no significant differences in clinical outcomes between patients who received random assignment of treatment (RCT participants) and those who received individualized treatment assignment (eligible non-participants). In addition, there was no relation between random treatment assignment and clinical outcome in 15 of the 17 studies (88%) in which randomized and nonrandomized patients had similar health status at baseline. CONCLUSIONS: These findings suggest that randomized treatment assignment as part of a clinical trial does not harm research participants. PMID- 16719554 TI - Unique author identification number in scientific databases: a suggestion. PMID- 16719552 TI - Fetal alcohol syndrome and essential fatty acids. PMID- 16719555 TI - Increasing the participation: another factor. PMID- 16719556 TI - The impact of open access upon public health. PMID- 16719557 TI - Improving imperfect data from health management information systems in Africa using space-time geostatistics. AB - BACKGROUND: Reliable and timely information on disease-specific treatment burdens within a health system is critical for the planning and monitoring of service provision. Health management information systems (HMIS) exist to address this need at national scales across Africa but are failing to deliver adequate data because of widespread underreporting by health facilities. Faced with this inadequacy, vital public health decisions often rely on crudely adjusted regional and national estimates of treatment burdens. METHODS AND FINDINGS: This study has taken the example of presumed malaria in outpatients within the largely incomplete Kenyan HMIS database and has defined a geostatistical modelling framework that can predict values for all data that are missing through space and time. The resulting complete set can then be used to define treatment burdens for presumed malaria at any level of spatial and temporal aggregation. Validation of the model has shown that these burdens are quantified to an acceptable level of accuracy at the district, provincial, and national scale. CONCLUSIONS: The modelling framework presented here provides, to our knowledge for the first time, reliable information from imperfect HMIS data to support evidence-based decision making at national and sub-national levels. PMID- 16719559 TI - Mechanisms of photoisomerization of polyenes in confined media: from organic glasses to protein binding cavities. AB - Photochemical reactivities of model organic systems (stilbene and diphenylbutadiene) in organic glasses were first examined and compared with those in solution and in organized media. These observations were in turn compared with reactivities of polyene chromophores in protein binding cavities (specifically PYP, rhodopsin and bacteriorhodopsin). The obvious conclusion is that the preference for the most volume-conserving Hula-twist mechanism isomerization in organic glasses is because of the close interaction between the guest and the host molecules. In organized media (zeolites, crystals and protein binding cavities), the residual empty space coupled with any specific guest-host interactions that are characteristic of a given system, could lead to involvement of the more volume-demanding one-bond-flip (i.e. torsional relaxation) or bicycle pedal or an extended HT process in photoisomerization. PMID- 16719560 TI - Resistance evolution to Bt crops: predispersal mating of European corn borers. AB - Over the past decade, the high-dose refuge (HDR) strategy, aimed at delaying the evolution of pest resistance to Bacillus thuringiensis (Bt) toxins produced by transgenic crops, became mandatory in the United States and is being discussed for Europe. However, precopulatory dispersal and the mating rate between resident and immigrant individuals, two features influencing the efficiency of this strategy, have seldom been quantified in pests targeted by these toxins. We combined mark-recapture and biogeochemical marking over three breeding seasons to quantify these features directly in natural populations of Ostrinia nubilalis, a major lepidopteran corn pest. At the local scale, resident females mated regardless of males having dispersed beforehand or not, as assumed in the HDR strategy. Accordingly, 0-67% of resident females mating before dispersal did so with resident males, this percentage depending on the local proportion of resident males (0% to 67.2%). However, resident males rarely mated with immigrant females (which mostly arrived mated), the fraction of females mating before dispersal was variable and sometimes substantial (4.8% to 56.8%), and there was no evidence for male premating dispersal being higher. Hence, O. nubilalis probably mates at a more restricted spatial scale than previously assumed, a feature that may decrease the efficiency of the HDR strategy under certain circumstances, depending for example on crop rotation practices. PMID- 16719561 TI - The obligate human pathogen, Neisseria gonorrhoeae, is polyploid. AB - We show using several methodologies that the Gram-negative, diplococcal-bacterium Neisseria gonorrhoeae has more than one complete genome copy per cell. Gene dosage measurements demonstrated that only a single replication initiation event per chromosome occurs per round of cell division, and that there is a single origin of replication. The region containing the origin does not encode any genes previously associated with bacterial origins of replication. Quantitative PCR results showed that there are on average three genome copies per coccal cell unit. These findings allow a model for gonococcal DNA replication and cell division to be proposed, in which a minimum of two chromosomal copies exist per coccal unit within a monococcal or diplococcal cell, and these chromosomes replicate in unison to produce four chromosomal copies during cell division. Immune evasion via antigenic variation is an important mechanism that allows these organisms to continually infect a high risk population of people. We propose that polyploidy may be necessary for the high frequency gene conversion system that mediates pilin antigenic variation and the propagation of N. gonorrhoeae within its human hosts. PMID- 16719563 TI - Empirical support for optimal virulence in a castrating parasite. AB - The trade-off hypothesis for the evolution of virulence predicts that parasite transmission stage production and host exploitation are balanced such that lifetime transmission success (LTS) is maximised. However, the experimental evidence for this prediction is weak, mainly because LTS, which indicates parasite fitness, has been difficult to measure. For castrating parasites, this simple model has been modified to take into account that parasites convert host reproductive resources into transmission stages. Parasites that kill the host too early will hardly benefit from these resources, while postponing the killing of the host results in diminished returns. As predicted from optimality models, a parasite inducing castration should therefore castrate early, but show intermediate levels of virulence, where virulence is measured as time to host killing. We studied virulence in an experimental system where a bacterial parasite castrates its host and produces spores that are not released until after host death. This permits estimating the LTS of the parasite, which can then be related to its virulence. We exposed replicate individual Daphnia magna (Crustacea) of one host clone to the same amount of bacterial spores and followed individuals until their death. We found that the parasite shows strong variation in the time to kill its host and that transmission stage production peaks at an intermediate level of virulence. A further experiment tested for the genetic basis of variation in virulence by comparing survival curves of daphniids infected with parasite spores obtained from early killing versus late killing infections. Hosts infected with early killer spores had a significantly higher death rate as compared to those infected with late killers, indicating that variation in time to death was at least in part caused by genetic differences among parasites. We speculate that the clear peak in lifetime reproductive success at intermediate killing times may be caused by the exceptionally strong physiological trade-off between host and parasite reproduction. This is the first experimental study to demonstrate that the production of propagules is highest at intermediate levels of virulence and that parasite genetic variability is available to drive the evolution of virulence in this system. PMID- 16719562 TI - Control of phage Bxb1 excision by a novel recombination directionality factor. AB - Mycobacteriophage Bxb1 integrates its DNA at the attB site of the Mycobacterium smegmatis genome using the viral attP site and a phage-encoded integrase generating the recombinant junctions attL and attR. The Bxb1 integrase is a member of the serine recombinase family of site-specific recombination proteins and utilizes small (<50 base pair) substrates for recombination, promoting strand exchange without the necessity for complex higher order macromolecular architectures. To elucidate the regulatory mechanism for the integration and excision reactions, we have identified a Bxb1-encoded recombination directionality factor (RDF), the product of gene 47. Bxb1 gp47 is an unusual RDF in that it is relatively large (approximately 28 kDa), unrelated to all other RDFs, and presumably performs dual functions since it is well conserved in mycobacteriophages that utilize unrelated integration systems. Furthermore, unlike other RDFs, Bxb1 gp47 does not bind DNA and functions solely through direct interaction with integrase-DNA complexes. The nature and consequences of this interaction depend on the specific DNA substrate to which integrase is bound, generating electrophoretically stable tertiary complexes with either attB or attP that are unable to undergo integrative recombination, and weakly bound, electrophoretically unstable complexes with either attL or attR that gain full potential for excisive recombination. PMID- 16719565 TI - Burnout and risk of cardiovascular disease: evidence, possible causal paths, and promising research directions. AB - Burnout is characterized by emotional exhaustion, physical fatigue, and cognitive weariness, resulting from prolonged exposure to work-related stress. The authors review the accumulated evidence suggesting that burnout and the related concept of vital exhaustion are associated with increased risk of cardiovascular disease and cardiovascular-related events. The authors present evidence supporting several potential mechanisms linking burnout with ill health, including the metabolic syndrome, dysregulation of the hypothalamic-pituitary-adrenal axis along with sympathetic nervous system activation, sleep disturbances, systemic inflammation, impaired immunity functions, blood coagulation and fibrinolysis, and poor health behaviors. The association of burnout and vital exhaustion with these disease mediators suggests that their impact on health may be more extensive than currently indicated. PMID- 16719564 TI - B cell antigen receptor signaling and internalization are mutually exclusive events. AB - Engagement of the B cell antigen receptor initiates two concurrent processes, signaling and receptor internalization. While both are required for normal humoral immune responses, the relationship between these two processes is unknown. Herein, we demonstrate that following receptor ligation, a small subpopulation of B cell antigen receptors are inductively phosphorylated and selectively retained at the cell surface where they can serve as scaffolds for the assembly of signaling molecules. In contrast, the larger population of non phosphorylated receptors is rapidly endocytosed. Each receptor can undergo only one of two mutually exclusive fates because the tyrosine-based motifs that mediate signaling when phosphorylated mediate internalization when not phosphorylated. Mathematical modeling indicates that the observed competition between receptor phosphorylation and internalization enhances signaling responses to low avidity ligands. PMID- 16719566 TI - Distributed practice in verbal recall tasks: A review and quantitative synthesis. AB - The authors performed a meta-analysis of the distributed practice effect to illuminate the effects of temporal variables that have been neglected in previous reviews. This review found 839 assessments of distributed practice in 317 experiments located in 184 articles. Effects of spacing (consecutive massed presentations vs. spaced learning episodes) and lag (less spaced vs. more spaced learning episodes) were examined, as were expanding interstudy interval (ISI) effects. Analyses suggest that ISI and retention interval operate jointly to affect final-test retention; specifically, the ISI producing maximal retention increased as retention interval increased. Areas needing future research and theoretical implications are discussed. PMID- 16719567 TI - Moving from cognition to behavior: What the research says. AB - In 1994, R. G. Lord and P. E. Levy proposed a variant of control theory that incorporated human information processing principles. The current article evaluates the empirical evidence for their propositions and updates the theory by considering contemporary research on information processing. Considerable support drawing from diverse literatures was found for propositions concerning the activation of goal-relevant information, the inhibition of goal-irrelevant information, and the consequences of goal completion. These effects were verified by meta-analytic analyses, which also supported the meaningfulness of such effects on the basis of their unstandardized magnitudes. The authors conclude by proposing new directions for this version of control theory by invoking recent theorizing on goal emergence and the importance of velocity and acceleration information for goal striving and by reviewing research in cognitive neuroscience. PMID- 16719568 TI - Polarity correspondence: A general principle for performance of speeded binary classification tasks. AB - Differences in performance with various stimulus-response mappings are among the most prevalent findings for binary choice reaction tasks. The authors show that perceptual or conceptual similarity is not necessary to obtain mapping effects; a type of structural similarity is sufficient. Specifically, stimulus and response alternatives are coded as positive and negative polarity along several dimensions, and polarity correspondence is sufficient to produce mapping effects. The authors make the case for this polarity correspondence principle using the literature on word-picture verification and then provide evidence that polarity correspondence is a determinant of mapping effects in orthogonal stimulus response compatibility, numerical judgment, and implicit association tasks. The authors conclude by discussing implications of this principle for interpretation of results from binary choice tasks and future model development. PMID- 16719569 TI - The addiction-stroop test: Theoretical considerations and procedural recommendations. AB - Decisions about using addictive substances are influenced by distractions by addiction-related stimuli, of which the user might be unaware. The addiction Stroop task is a paradigm used to assess this distraction. The empirical evidence for the addiction-Stroop effect is critically reviewed, and meta-analyses of alcohol-related and smoking-related studies are presented. Studies finding the strongest effects were those in which participants had strong current concerns about an addictive substance or such concerns were highlighted through experimental manipulations, especially those depriving participants of the substance. Theories to account for addiction-related attentional bias are discussed, of which the motivational theory of current concerns appears to provide the most complete account of the phenomenon. Recommendations are made for maximizing the precision of the addiction-Stroop test in future research. PMID- 16719571 TI - Prediction of prosocial and emotional competence from maternal behavior in African American preschoolers. AB - This research investigates whether individual differences in African American preschoolers' prosocial behavior and emotional regulation ability were differentially predicted by parenting practices around prosocial behavior and emotions, once variance due to SES was explained. Seventy African American preschoolers were visited at their preschools and their prosocial and emotion regulation behaviors were observed. Mothers' emotion and prosocial-related socialization behaviors were observed during a separate home visit. Positive prosocial-related maternal variables (i.e., social approval of the act and praise of the child) were positively associated with prosocial behavior, above and beyond SES. On the other hand, positive emotion-related maternal variables, such as matching and discussion of emotions were more strongly related to emotion regulation behavior. Findings are discussed in terms of the lack of research on higher SES African American children and the associated methodological challenges of conducting developmental research on diverse populations. PMID- 16719572 TI - Constructions of Mexican American family grief after the death of a child: an exploratory study. AB - Using a collective case study ethnographic approach, nine individuals comprising three Mexican American families were interviewed about their family bereavement experiences after the death of a child. All families were Catholic, had surviving siblings, and had had three or more years to grieve their loss when interviewed. The deceased children ranged in age from 3 to 14, and all experienced sudden, traumatic, nonsuicide deaths. To provide a broader, contextual picture of their grief experiences, four individuals who supported these family members after the loss were also interviewed. Unique grief experiences were identified, and eight common themes emerged, reflecting the ways in which family members maintained their bond to the deceased: dreams, storytelling, keepsakes, sense of presence, faith-based connections, proximity connections, ongoing rituals, and pictorial remembrances. The cultural implications of family bereavement are highlighted. PMID- 16719573 TI - The relationships among trauma, stress, ethnicity, and binge eating. AB - The present study investigated whether trauma, stress, and discriminatory experiences influenced binge eating among 93 African American and 85 Caucasian women. Trauma and stress were significantly related to binge eating for both groups, although the stress- binge eating relationship was stronger for Caucasian women. Ethnicity did not moderate the relationship between trauma and binge eating, but did moderate the stress-binge eating relationship. Finally, the hypothesis that trauma and stress would influence binge eating through their effects on function of eating was partially supported; the relationship between stress and binge eating was partially mediated by function of eating among Caucasian women. The implications of these findings for our understanding of binge eating are discussed. PMID- 16719570 TI - Inventorying stressful life events as risk factors for psychopathology: Toward resolution of the problem of intracategory variability. AB - An explosion of research on life events has occurred since the publication of the Holmes and Rahe checklist in 1967. Despite criticism, especially of their use in research on psychopathology, such economical inventories have remained dominant. Most of the problems of reliability and validity with traditional inventories can be traced to the intracategory variability of actual events reported in their broad checklist categories. The purposes of this review are, first, to examine how this problem has been addressed within the tradition of economical checklist approaches; second, to determine how it has been dealt with by far less widely used and far less economical labor-intensive interview and narrative-rating approaches; and, third, to assess the prospects for relatively economical, as well as reliable and valid, solutions. PMID- 16719574 TI - Parent-child value transmission among Chinese immigrants to North America: the case of traditional mate preferences. AB - Parent-child similarities in traditional mate preferences were examined in Chinese immigrants to North America. Adult children (n = 63) rated their preferred mate characteristics. Children then completed measures of interdependence, family allocentrism, Chinese identity, and Asian values. Their parents (n = 63) also rated their own preferences for their child's spouse and their own Asian values. Parents reported greater preference for traditional mate characteristics than did their children. Parents' preferences were related to parents' own Asian values. Children's traditional mate preferences were predicted by their parents' preferences and their own family allocentrism. Family allocentrism was a marginally significant partial mediator of parent's influence on children's preferences. Family connectedness may, therefore, facilitate intergenerational transmission of values in immigrant Asian families. PMID- 16719575 TI - Behavioral acculturation and enculturation and psychological functioning among Asian American college students. AB - The purpose of this study was to examine Asian American college students' behavioral acculturation to U.S. cultural norms and behavioral enculturation to Asian cultural norms and their relationships to ratings on measures of cognitive flexibility, general self-efficacy, collective self-esteem, acculturative stress, and attitudes toward seeking professional psychological help. As hypothesized, the results indicated that both acculturation and enculturation were positively related to the membership dimension of collective self-esteem. Although not hypothesized, the results indicated that acculturation was positively related to cognitive flexibility, general self-efficacy, and the public dimension of collective self-esteem. In addition, enculturation was positively related to the private dimension and the importance of identity dimension of collective self esteem. PMID- 16719576 TI - Asian American college women's body image: a pilot study. AB - The purpose of this study was to examine the effects of values acculturation and the influence of media on Asian American college women's overall body image. A sample of 59 Asian American women from two large universities completed self report surveys, which included questions regarding values acculturation, media internalization, and overall body satisfaction. Results showed that Asian American women who identified more strongly with traditional Asian values reported higher levels of body image dissatisfaction. Further, Asian American women who reported higher internalization of media portrayals of beauty ideals reported higher body image dissatisfaction. Research and clinical recommendations are made to enhance psychologists' understanding of Asian American women's body image and acculturation. PMID- 16719577 TI - Exploring the association between color-blind racial ideology and multicultural counseling competencies. AB - The authors examined the association between color-blind racial ideology and self reported multicultural counseling competencies in 130 applied psychology students and mental health workers. Results from 1 sample (n = 79) indicated that greater levels of color-blind racial ideology as measured by the Color-Blind Racial Attitudes Scale (Neville, Lilly, Duran, Lee, & Browne, 2000) were (1) related to lower self-reported multicultural counseling awareness and knowledge as measured by the Multicultural Counseling Knowledge and Awareness Scale (MCKAS; Ponterotto, Gretchen, Utsey, Rieger, & Austin, 2002) and (2) accounted for a significant amount of variance in MCKAS scores over and above that explained by self-reported multicultural training, social desirability, and participant race. Findings from another sample also provided empirical support for the link between higher color blind racial ideology and lower multicultural case conceptualization ability (n = 51), after controlling for the influence of the number of multicultural course(s) taken. Implications of the findings and future directions are provided. PMID- 16719579 TI - South Asian women: identities and conflicts. AB - This study investigated the effects of education, level of religiosity, ethnic identity (internal and external), and racial identity statuses (conformity, dissonance, resistance, and awareness) on cultural value conflict for first and second generation South Asian women (N = 193). Cultural value conflict was examined in two areas, intimate relations and sex-role expectations. Results revealed that level of religiosity was more predictive of intimate relations conflict for second generation than first generation women. Furthermore, higher ratings in internal ethnic identity predicted greater intimate relations conflict in first generation women, while both internal and external ethnic identity predicted intimate relations conflict in second generation women. Finally, higher dissonance scores in racial identity predicted greater conflicts in sex-role expectations for second generation women alone. PMID- 16719578 TI - Frequency and intensity of social anxiety in Asian Americans and European Americans. AB - Asian American students have typically reported greater levels of social anxiety than European American students on self-report measures (e.g., Okazaki, 1997; Norasakkunkit & Kalick, 2002). This study employed an event-contingent experience sampling methodology to examine whether Asian American university students experienced social anxiety more often and more intensely than European Americans in their daily lives. Forty-five Asian American and 38 European American students participated in a two-week diary study. The results showed that on average, Asian Americans and European Americans reported a similar number of events that evoked anxiety in social situations, but Asian Americans reported more negative emotions on average in social situations than did European Americans. PMID- 16719580 TI - A cultural-contextual perspective on the validity of the MMPI-2 with American Indians. AB - This study investigated the normative validity of the MMPI-2 with two distinct American Indian tribes. Differences occurred on 8 of the 13 basic validity and clinical scales (F, 1, 4, 5, 6, 7, 8, 9) between the MMPI-2 norms and both tribal samples. Elevated MMPI-2 scores of American Indians may not only reflect the possibility of psychological distress spurred by historical oppression and present adversity, but also an expression of a divergent worldview. Considering the context of the historical and social production of knowledge about American Indians, it is argued that researchers and practitioners, when interpreting MMPI 2 results for American Indians, should seriously consider their interpretive points of reference, which may be impacted by dominant cultural belief systems. PMID- 16719581 TI - Moderator effects of cognitive ability and social support on the relation between race-related stress and quality of life in a community sample of black Americans. AB - The current study examined the combined moderating effects of cognitive ability and social support on the relation between race-related stress and quality of life in a sample of Black Americans. Participants (N = 323) were administered the Wonderlic Personnel Test (WPT; E. F. Wonderlic Associates, Inc., 1983), the Multidimensional Social Support Scale (MDSS; Winefield, Winefield, & Tiggemann, 1992), the Index of Race-Related Stress-Brief (Utsey, 1999), and the WHOQOL-BREF (The WHO Group, 1998). The findings indicated that cognitive ability and social support, conjointly, moderated the relation between individual and cultural race related stress and quality of life for Black Americans in the current sample. The paper concludes by discussing the study's findings, limitations, and by offering recommendations for future research related to this area of inquiry. PMID- 16719582 TI - Reasons to attend college among ethnically diverse college students. AB - A survey of 713 ethnically diverse university freshmen (463 Latino, 167 Asian American, 54 African American, 29 European American) assessed reasons for attending college, ethnic identity, family interdependence, and college adjustment. Results revealed three reasons for attending college not reported in previous research: to help one's family, to prove one's self-worth, and because of encouragement. Attending college to help the family was stronger among students from lower socioeconomic status backgrounds. Ethnic identity and family interdependence contributed positively to some reasons for attending college, but did not mediate ethnic differences; with cultural factors and socioeconomic status included as predictors, ethnic minority students still gave greater importance to family oriented motives than did European Americans. Career/personal and humanitarian motives contributed positively to college adjustment. PMID- 16719583 TI - A cross-cultural analysis of the Brief Symptom Inventory-18. AB - The Brief Symptom Inventory-18 (BSI-18: Derogatis, 2000) is an abbreviated version of the nine dimension, 53-item BSI (Derogatis, 1993) which is a shortened form of the 90-item Symptom Checklist-90-Revised (SCL-90-R: Derogatis, 1994). Criticism focused on factor structure (cf. Boulet & Boss, 1991; Ruiperez, Ibanez, Lorente, Moro, & Ortet, 2001) and the two older versions' poor discriminant validity suggest the scales' usefulness is limited to global scores only. Using principal component analysis, the authors explore the three-dimension factor structure of the BSI - 18 with the understudied population of Central American immigrants to the U.S. (N = 100). Results suggest one underlying factor best used in aggregate as a general screening for overall levels of psychological distress. Cultural differences are discussed. PMID- 16719584 TI - Editorial: Journal of Comparative Psychology. AB - Both continuity and change typically mark the changing of editors at a long established journal with extended editorial terms. Change is inherent in any dynamic field and is independent of editorship, but editors have an influence that should be wielded in a fair, responsible, judicious, and scientifically rigorous manner, while inevitably reflecting their own perspectives and values. The Journal of Comparative Psychology will continue to publish exciting, fascinating, assessable, controversial, and well-written reports on research, be the topic traditional, interdisciplinary, applied, or one breaking risky new ground. Editorial standards must be high, but appropriate for various subfields, and as editor the author will try to make those judgments carefully. The author would also like to see more submissions of brief reports describing exciting developments as well as submissions on significant theoretical, conceptual, and methodological issues during his tenure as editor. PMID- 16719585 TI - Song perception during the sensitive period of song learning in zebra finches (Taeniopygia guttata). AB - The sensitive period is a special time for auditory learning in songbirds. However, little is known about perception and discrimination of song during this period of development. The authors used a go/no-go operant task to compare discrimination of conspecific song from reversed song in juvenile and adult zebra finches (Taeniopygia guttata), and to test for possible developmental changes in perception of syllable structure and syllable syntax. In Experiment 1, there were no age or sex differences in the ability to learn the discrimination, and the birds discriminated the forward from reversed song primarily on the basis of local syllable structure. Similar results were found in Experiment 2 with juvenile birds reared in isolation from song. Experiment 3 found that juvenile zebra finches could discriminate songs on the basis of syllable order alone, although this discrimination was more difficult than one based on syllable structure. The results reveal well-developed song discrimination and song perception in juvenile zebra finches, even in birds with little experience with song. PMID- 16719586 TI - Do chimpanzees (Pan troglodytes) and 2-year-old children (Homo sapiens) understand double invisible displacement? AB - Chimpanzees (Pan troglodytes) and young children (Homo sapiens) have difficulty with double invisible displacements in which an object is hidden in two nonadjacent boxes in a linear array. Experiment 1 eliminated the possibility that chimpanzees' previous poor performance was due to the hiding direction of the displacement device. As in Call (2001), subjects failed double nonadjacent displacements, showing a tendency to select adjacent boxes. In Experiments 2 and 3, chimpanzees and 24-month-old children were tested on a new adaptation of the task in which four hiding boxes were presented in a diamond-shaped array on a vertical plane. Both species performed above chance on double invisible displacements using this format, suggesting that previous poor performance was due to a response bias or inhibition problem rather than a fundamental limitation in representational capacity. PMID- 16719587 TI - Behavioral limits of auditory temporal resolution in the rat: amplitude modulation and duration discrimination. AB - Thresholds for detecting the presence of amplitude modulation in a noise carrier were determined for rats using conditioned avoidance procedures. There was a progressive increase in threshold with modulation rates between 5 Hz and 2 kHz. Further tests were conducted to determine difference thresholds for detecting an increase in modulation rate for standard rates of 10, 50, and 100 Hz. The size of the difference threshold increased progressively as the standard rate increased. In addition, thresholds for detecting an increase in the duration of a noise burst were determined for various standard durations. The difference thresholds were constant for values between 10 and 50 ms but increased progressively, with standard durations between 0.1 and 1.0 s. PMID- 16719589 TI - Do dogs (Canis familiaris) seek help in an emergency? AB - The question of whether dogs recognize an emergency and understand the need to seek help from a bystander was tested in two experiments. In the first experiment, dogs' owners feigned a heart attack in an open field, and in the second experiment, dogs' owners experienced an accident in which a bookcase fell on them and pinned them to the floor. In these experiments, one or two bystanders were available to which dogs could go for help. The dogs' behavior was taped for 6 min after the owner had fallen and was later scored for the frequency and time the dogs spent performing different behaviors. In no case did a dog solicit help from a bystander. It is concluded that dogs did not understand the nature of the emergency or the need to obtain help. PMID- 16719588 TI - Sex-specific differences in olfactory sensitivity for putative human pheromones in nonhuman primates. AB - In humans, the volatile C19-steroids androsta-4,16-dien-3-one (AND) and estra 1,3,5(10),16-tetraen-3-ol (EST) have been shown to modulate autonomic nervous system responses, and to cause hypothalamic activation in a gender-specific manner. Using two conditioning paradigms, the authors here show that pigtail macaques and squirrel monkeys of both sexes were able to detect AND and EST at concentrations in the micromolar and mM range, respectively. Male and female spider monkeys, in contrast, differed markedly in their sensitivity to these two odorous steroids, with males not showing any behavioral responses to the highest concentrations of AND tested and females not responding to the highest concentrations of EST. These data provide the first examples of sex-specific bimodal distributions of olfactory sensitivity in a nonhuman primate species. PMID- 16719590 TI - Geometrical gaze following in common marmosets (Callithrix jacchus). AB - A series of experiments investigating the degree of gaze understanding in common marmosets (Callithrix jacchus) is reported. Results show that marmosets follow the gaze of a human experimenter readily and also use the gaze to locate food in a modified version of the object choice task if influences of chance probabilities and prepotent response tendencies are controlled for. In addition, this new version of the task allows the assessment of the accuracy of gaze following. Marmosets precisely extrapolate gaze direction, past distracting objects and from considerable distances, thereby meeting the criteria of so called geometrical gaze following. The presence of this ability in common marmosets suggests that higher forms of gaze following might be more widely distributed among nonhuman primates than previously thought. PMID- 16719592 TI - Comparative evolutionary psychology of sperm competition. AB - A comparative evolutionary psychological perspective predicts that species that recurrently faced similar adaptive problems may have evolved similar psychological mechanisms to solve these problems. Sperm competition provides an arena in which to assess the heuristic value of such a comparative evolutionary perspective. The sperm competition that results from female infidelity and polyandry presents a similar class of adaptive problems for individuals across many species. The authors first describe mechanisms of sperm competition in insects and in birds. They suggest that the adaptive problems and evolved solutions in these species provide insight into human anatomy, physiology, psychology, and behavior. The authors then review recent theoretical and empirical arguments for the existence of sperm competition in humans and discuss proposed adaptations in humans that have analogs in insects or birds. The authors conclude by highlighting the heuristic value of a comparative evolutionary psychological approach in this field. PMID- 16719591 TI - The relative importance of global and local landmarks in navigation by Columbian ground squirrels (Spermophilus columbianus). AB - In order to survive, small burrowing mammals need to remember the locations of escape burrows. Therefore, it is important to know what types of landmarks are used to aid navigation in the wild. The author tested the ability of free-ranging Columbian ground squirrels (Spermophilus columbianus) to locate escape burrows when local (e.g., vegetation pattern, local relief), global (e.g., forest edge, mountain outline), or both types of landmarks were obstructed. Results suggest that squirrels need both local and global landmarks of the environment for successful navigation, and that the upper portion of the horizon is especially important for orientation. Moreover, the lack of information from one type of landmark (local or global) cannot be completely compensated by the other type. PMID- 16719593 TI - Statistical classification of black-capped (Poecile atricapillus) and mountain chickadee (Poecile gambeli) call notes. AB - Both black-capped (Poecile atricapillus) and mountain chickadees (Poecile gambeli) produce a chick-a-dee call that consists of several distinct note types. In some regions, these 2 species live sympatrically, and it has been shown that 1 species will respond weakly to songs of the other. This suggests that chickadee song, and potentially other of their vocalizations, contains species-specific information. We tested the possibility that call notes were acoustically sufficient for species identification. Black-capped and mountain non-D notes were summarized as a set of 9 features and then analyzed by linear discriminant analysis. Linear discriminant analysis was able to use these notes to identify species with 100% accuracy. We repeated this approach, but with black-capped and mountain D notes that were summarized as a set of 4 features. Linear discriminant analysis was able to use these notes to identify species with 94% accuracy. This demonstrates that any of the note types in these chickadee calls possesses sufficient information for species classification. PMID- 16719594 TI - Chimpanzees (Pan troglodytes) conceal visual and auditory information from others. AB - Chimpanzees (Pan troglodytes) competed with a human for food. The human sat inside a booth, with 1 piece of food to her left and 1 to her right, which she could retract from her chimpanzee competitor's reach as needed. In Experiment 1, chimpanzees could approach either side of the booth unseen but then had to reach through 1 of 2 tunnels (1 clear, 1 opaque) for the food. In Experiment 2, both tunnels were clear and the human was looking away, but 1 of the tunnels made a loud noise when it was opened. Chimpanzees preferentially reached through the opaque tunnel in the first study and the silent tunnel in the second, successfully concealing their taking of the food from the human competitor in both cases. These results suggest that chimpanzees can, in some circumstances, actively manipulate the visual and auditory perception of others by concealing information from them. PMID- 16719595 TI - Self-control and tool use in tufted capuchin monkeys (Cebus apella). AB - Self-control is defined as forgoing immediate gratification to obtain a greater reward. Tool use may relate to self-control because both behaviors may require foresight and deliberate control over one's actions. The authors assessed 20 capuchin monkeys (Cebus apella) for the ability to delay gratification in a tool task. Subjects were given rod-shaped food items that could either be consumed immediately or be carried to an apparatus and used to extract a more preferred food. The authors found that some monkeys were able to exhibit self-control. Monkeys with relatively more tool use experience demonstrated the greatest levels of self-control. These results indicate that capuchins are capable of delaying gratification when a higher quality reinforcer is present and that tool experience can influence levels of self-control in this task. PMID- 16719596 TI - Medical students' and residents' gender bias in the diagnosis, treatment, and interpretation of coronary heart disease symptoms. AB - Two competing hypotheses explaining gender bias in cardiac care were tested. The first posits that women's coronary heart disease (CHD) symptoms are simply misinterpreted or discounted. The second posits that women's CHD symptoms are misinterpreted when presented in the context of stress. In two studies, medical students and residents randomized to 2 (male vs. female) x 2 (stress vs. nostress) experiments read vignettes of patients with CHD symptoms and indicated their diagnosis, treatment, and symptom origin interpretation. Both studies disconfirmed the first hypothesis and strongly supported the second. Only when stress was added did women receive significantly lower CHD diagnoses and cardiologist referrals than men and did the origin interpretation of women's CHD symptoms (e.g., chest pain) shift from organic to psychogenic. Neither participants' gender nor their attitude toward women influenced assessments. PMID- 16719597 TI - Relaxation versus cognitive therapies for dental fear--a psychophysiological approach. AB - By separating 127 adult dental-phobic patients according to fear etiology and psychophysiologic response style, the authors investigated the outcome of 2 dental fear treatments. Before and after either relaxation or cognitively oriented treatment, subjects were exposed to neutral and fear-relevant video sequences while the subjects' forehead muscle tension, heart rate, and skin conductance were recorded. Pre- to postintervention differences in self-reported dental fear, general fear, and trait anxiety were analyzed together with psychophysiological data. Both treatments resulted in a significant reduction of dental fear. Despite significant interaction effects of Treatment Modality x Psychophysiological Response Style, it could not be concluded that patients with different fear etiologies or response styles benefit differentially from the 2 treatments given. PMID- 16719599 TI - The impact of self-efficacy on behavior change and weight change among overweight participants in a weight loss trial. AB - Despite considerable clinical interest, attempts to link perceived self-efficacy with successful weight control have had mixed success. Definitive data on prospective associations between self-efficacy and weight loss are particularly sparse. This study examined relationships between self-efficacy beliefs, weight control behaviors, and weight change among individuals participating in a weight loss trial (N = 349, 87% women). Cross-sectionally, eating and exercise self efficacy beliefs were strongly associated with corresponding weight loss behaviors. Self-efficacy beliefs prospectively predicted weight control behavior and weight change during active treatment but not during follow-up. Mediational models indicate that people's weight control behaviors mediate the impact of self efficacy on weight change. PMID- 16719598 TI - Relation of successful dietary restriction to change in bulimic symptoms: a prospective study of adolescent girls. AB - Recent experimental evidence that dietary restriction results in decreased bulimic and depressive symptoms seems inconsistent with findings from prospective studies and etiologic theory. However, because the dieting manipulated in these experiments may be unrepresentative of real-world weight loss dieting, the authors tested whether successful dietary restriction was associated with decreases in these outcomes by using longitudinal data from a school-based study of 496 adolescent girls. Moderately overweight participants who evidenced successful dietary restriction showed significantly greater decreases in bulimic symptoms than weight-matched participants who did not show successful dietary restriction; however, there were no effects for depressive symptoms. In conjunction with past experimental findings, results seem to imply that successful dietary restriction curbs bulimic symptoms, suggesting that current etiologic models may need revision. PMID- 16719600 TI - Understanding action control: predicting physical activity intention-behavior profiles across 6 months in a Canadian sample. AB - Intention is theorized as the proximal determinant of behavior in some theories of motivation, but the need to understand predictors of action control (i.e., translating an intention into behavior) is warranted to tailor physical activity intervention efforts. The purpose of this study was to examine constructs of the transtheoretical model of behavior change (TTM) as predictors of physical activity intention-behavior profiles across 6 months in a large Canadian sample (N = 1,192). Results showed that 5 of the 8 possible intention-behavior profiles had a substantial number of participants: nonintenders, unsuccessful adopters, successful adopters, unsuccessful maintainers, and successful maintainers. Constructs of the TTM distinguished (p < .01) intention-behavior profiles. Self efficacy and the behavioral processes of change were particularly good predictors of action control (p < .01), but disaggregated beliefs and processes identified specific intervention targets for successful physical activity adoption and maintenance. The results validate that both action planning and action control are important when understanding physical activity behavior. PMID- 16719601 TI - Increased attention for computer-tailored health communications: an event-related potential study. AB - The authors tested whether individually tailored health communications receive more attention from the reader than nontailored health communications in a randomized, controlled trial among student volunteers (N = 24). They used objective measures of attention allocation during the message exposure. In a between-subjects design, participants had to read tailored or nontailored nutrition education messages and at the same time had to pay attention to specific odd auditory stimuli in a sequence of frequent auditory stimuli (odd ball paradigm). The amount of attention allocation was measured by recording event-related potentials (ERPs; i.e., N100 and P300 ERPs) and reaction times. For the tailored as opposed to the nontailored group, results revealed larger amplitudes for the N100 effect, smaller amplitudes for the P300 effect, and slower reaction times. Resource allocation theory and these results suggest that those in the tailored group allocated more attention resources to the nutrition message than those in the nontailored group. PMID- 16719602 TI - Randomized clinical trial of local anesthetic versus a combination of local anesthetic with self-hypnosis in the management of pediatric procedure-related pain. AB - A prospective controlled trial was conducted to compare the efficacy of an analgesic cream (eutectic mixture of local anesthetics, or EMLA) with a combination of EMLA with hypnosis in the relief of lumbar puncture-induced pain and anxiety in 45 pediatric cancer patients (age 6-16 years). The study also explored whether young patients can be taught and can use hypnosis independently as well as whether the therapeutic benefit depends on hypnotizability. Patients were randomized to 1 of 3 groups: local anesthetic, local anesthetic plus hypnosis, and local anesthetic plus attention. Results confirmed that patients in the local anesthetic plus hypnosis group reported less anticipatory anxiety and less procedure-related pain and anxiety and that they were rated as demonstrating less behavioral distress during the procedure. The level of hypnotizability was significantly associated with the magnitude of treatment benefit, and this benefit was maintained when patients used hypnosis independently. PMID- 16719603 TI - Causal ordering of physical self-concept and exercise behavior: reciprocal effects model and the influence of physical education teachers. AB - Does prior physical self-concept influence subsequent exercise behavior? On the basis of a large sample of physical education classes (2,786 students, 200 classes, 67 teachers) collected early (Time 1) and late (Time 2) in the school year, findings support a reciprocal effects model in which prior physical self concept and exercise behavior both influence subsequent physical self-concept and exercise behavior. Whereas variables from the theory of planned behavior (TOPB; behavioral intentions, perceived behavioral control, exercise attitudes) also contributed to the prediction of subsequent exercise behavior, the effect of prior physical self-concept was significant for subsequent outcomes after controlling these variables, suggesting that the TOPB should be supplemented with self-concept measures. On the basis of multilevel models, there were systematic differences in these variables for students taught by different teachers that generalized over time and across different classes taught by the same teacher. Support for the reciprocal effects model was robust. PMID- 16719604 TI - Adherence to antiretroviral medications in HIV: differences in data collected via self-report and electronic monitoring. AB - Controversy remains regarding the reliability of methods used to determine adherence to antiretroviral medication in HIV. In this study the authors compared adherence rates of 119 HIV-positive participants during a 6-month study, as estimated via electronic monitoring (EM) and self-report (SR). Adherence for both short (4-day) and long (4-week, or intervisit) periods was examined, as well as factors that underlie discrepancies between EM and SR. Results showed that intervisit EM estimates were consistently lower than those of SR. SR estimates based on shorter periods (4 days) were closer to those of EM. Higher discrepancies between EM and SR estimates were associated with lower cognitive functioning and externalized locus of control. These findings lend support for using both EM and SR methods; however, study design (e.g., length) and other factors (e.g., cognitive status, cost) should be considered. PMID- 16719605 TI - Identifying subgroups that succeed or fail with three levels of physical activity intervention: the Activity Counseling Trial. AB - The authors used recursive partitioning methods to identify combinations of baseline characteristics that predict 2-year physical activity success in each of 3 physical activity interventions delivered in the multisite Activity Counseling Trial. The sample consisted of 874 initially sedentary primary care patients, ages 35-75 years, who were at risk for cardiovascular disease. Predictors of 2 year success were specific to each intervention and represented a range of domains, including physiological, demographic, psychosocial, health-related, and environmental variables. The results indicate how specific patient subgroups (e.g., obese, unfit individuals; high-income individuals in stable health) may respond differently to varying levels and amounts of professional assistance and support. The methods used provide a practical first step toward identifying clinically meaningful patient subgroups for further systematic investigation. PMID- 16719606 TI - Moderators of the benefits of psychoeducational interventions for men with prostate cancer. AB - The authors examined whether 3 individual difference variables--self-esteem, self efficacy, and depressive symptoms--interacted with psychoeducational group interventions for men with prostate cancer (n = 250) to predict general and prostate-specific quality of life. Men with nonmetastatic prostate cancer were randomly assigned to an education intervention, an education plus group discussion intervention, or usual care and followed for 12 months. Eight groups of 10 to 12 men were convened in each condition. Men who began the study with lower self-esteem, lower prostate-specific self-efficacy, and higher depressive symptoms benefited the most from the interventions. Of these 3 moderator variables, the most consistent results emerged for self-esteem. That is, the benefits of the intervention were strongest for men with low self-esteem. PMID- 16719608 TI - Perceived racism and cardiovascular reactivity and recovery to personally relevant stress. AB - This study evaluated cardiovascular responses (CVR) to an active speech task with blatantly discriminatory (BRC) versus neutral (NRC) stimuli and an anger recall task in a sample of Black men (N = 73; age 18 to 47). Diastolic blood pressure scores were higher for NRC versus BRC stimuli during anger recall (p = .05). Moreover, persons in the NRC group who perceived high levels of racism (vs. no racism or BRC group) during active speech showed larger increases in blood pressure across postspeech rest, anger recall, and subsequent rest (p = .03). The notable elevation in CVR in response to an ambiguous event extends current models of racism suggesting that subtle racism is a psychosocial stressor that erodes health through chronically elevated CVR. PMID- 16719607 TI - A Social Problem-Solving Model of Adherence to HIV Medications. AB - HIV medication adherence remains a challenge and limits the degree to which treatment benefit can be maximized. This study tested an explanatory model of HIV medication adherence using a social problem-solving (SPS) framework. Associations of SPS with adherence are hypothesized to be direct and/or indirect via psychological health. HIV+ adults were interviewed using validated measures of SPS, psychological health, and antiretroviral therapy (ART) medication adherence. Structural equation modeling (SEM) techniques were used to test hypothesized relationships and to evaluate overall fit of the model to the data. SEM supported an indirect association (but not direct) of SPS on adherence via psychological health among the 545 HIV+ adults included in the analyses. Overall, the findings resulted in a model of adherence that offered very good fit to the data and correctly classified 97% of the cases as adherent versus nonadherent. Results support the use of SPS as a conceptual framework for understanding adherence to ART. Findings offer rationale and direction for SPS interventions to enhance adherence by improving psychological health. Such approaches, if effective, have the potential to positively impact psychological well being and adherence, thereby maximizing clinical benefit from treatment, which is linked to lower mortality from AIDS. PMID- 16719609 TI - Depression history and coping with chronic pain: a daily process analysis. AB - This study examined how a previous episode of depression is related to daily pain and reactions to pain among individuals with fibromyalgia, a chronic pain syndrome. Seventy-one women with fibromyalgia (including 30 who were previously depressed) rated their pain and mood 3 times daily for 30 days. Each night, participants rated the extent to which they responded to pain by catastrophizing, how much control they had over that day's pain, their ways of coping with pain that day, and the effectiveness of their coping efforts. Multivariate multilevel regression models revealed that after controlling for neuroticism and current depressive symptoms, formerly depressed and never-depressed individuals differed in how they coped with increased pain and in how they appraised the efficacy of their coping efforts. Formerly depressed participants who also reported more current depressive symptoms showed a greater decline in pleasant mood on more painful days than did formerly depressed participants who were experiencing fewer current depressive symptoms. These findings illustrate how a history of depression can be captured in the dynamics of daily life. PMID- 16719610 TI - Comparing intervention outcomes in smokers treated for single versus multiple behavioral risks. AB - This study examined treatment outcomes among smokers with single versus multiple behavioral risks. Data were drawn from smokers (N = 2,326) participating in 3 population-based, multibehavioral studies targeting cigarette smoking, high-fat diets, and high-risk sun exposure. Outcomes were compared for participants at risk for smoking only (13%), smoking plus 1 additional risk factor (37%), and smoking plus 2 additional risk factors (50%). The smoking only group tended to be female, older, more educated, lighter smokers, in the preparation stage of change for smoking, with more previous quit attempts and longer periods of abstinence. At 12 and 24 months follow-up, treatment of 1 or 2 coexisting risk factors did not decrease the effectiveness of smoking cessation treatment, and treatment for the coexisting factors was effective. PMID- 16719611 TI - Symptom perception in childhood asthma: the role of anxiety and asthma severity. AB - This study tested the relationship of anxiety and asthma severity to symptom perception. Eighty-six children diagnosed with mild or moderate asthma had symptom perception and pulmonary function measured throughout methacholine challenge (to induce bronchoconstriction). Higher trait anxiety was associated with heightened symptom perception (controlling for pulmonary function) at baseline. Greater asthma severity was associated with blunted symptom perception (controlling for pulmonary function) at the end of methacholine challenge and with a slower rate of increase in symptom perception across methacholine challenge. These results suggest that anxiety plays a role when children's symptoms are mild, whereas medical variables such as severity play a role in perception of changes in asthma symptomatology as bronchoconstriction worsens. PMID- 16719612 TI - Physical self-concept and self-esteem mediate cross-sectional relations of physical activity and sport participation with depression symptoms among adolescent girls. AB - The authors tested whether physical self-concept and self-esteem would mediate cross-sectional relations of physical activity and sport participation with depression symptoms among 1,250 girls in 12th grade. There was a strong positive relation between global physical self-concept and self-esteem and a moderate inverse relation between self-esteem and depression symptoms. Physical activity and sport participation each had an indirect, positive relation with global physical self-concept that was independent of objective measures of cardiorespiratory fitness and body fatness. These correlational findings provide initial evidence suggesting that physical activity and sport participation might reduce depression risk among adolescent girls by unique, positive influences on physical self-concept that operate independently of fitness, body mass index, and perceptions of sports competence, body fat, and appearance. PMID- 16719613 TI - Rail commuting duration and passenger stress. AB - Over 100 million Americans commute to work every weekday. Little is known, however, about how this aspect of work, which may indeed be the most stressful aspect of the job for some, affects human health and well-being. The authors studied a sample of 208 male and female suburban rail commuters who took the train to Manhattan, New York. The greater the duration of the commute, the larger the magnitude of salivary cortisol elevations in reference to resting baseline levels, the less the commuter's persistence on a task at the end of the commute, and the greater the levels of perceived stress. These effects were not moderated by gender. Commuting stress is an important and largely overlooked aspect of environmental health. PMID- 16719614 TI - A path analysis of factors associated with distress among first-degree female relatives of women with breast cancer diagnosis. AB - Patterns and predictors of psychological distress in first-degree female relatives (N = 624) of newly diagnosed breast cancer patients were explored. First-degree female relatives who were high monitors reported greater cancer specific and general distress than did low monitors. Greater optimism was associated with lower cancer-specific distress. Optimism's effect on general distress was moderated by women's level of monitoring. Greater optimism was associated with lower general distress for both high and low monitors, but the effect was stronger for high monitors than for low monitors. Avoidance and engaged coping were associated with higher distress. A close relationship with the cancer patient was related to higher cancer-specific distress but lower general distress. Further understanding of the process of adjustment in these women awaits longitudinal study. PMID- 16719615 TI - How well do people recall risk factor test results? Accuracy and bias among cholesterol screening participants. AB - The authors conducted a community-based cholesterol screening study to examine accuracy of recall for self-relevant health information in long-term autobiographical memory. Adult community residents (N = 496) were recruited to participate in a laboratory-based cholesterol screening and were also provided cholesterol counseling in accordance with national guidelines. Participants were subsequently interviewed 1, 3, or 6 months later to assess their memory for their test results. Participants recalled their exact cholesterol levels inaccurately (38.0% correct) but their cardiovascular risk category comparatively well (88.7% correct). Recall errors showed a systematic bias: Individuals who received the most undesirable test results were most likely to remember their cholesterol scores and cardiovascular risk categories as lower (i.e., healthier) than those actually received. Recall bias was unrelated to age, education, knowledge, self rated health status, and self-reported efforts to reduce cholesterol. The findings provide evidence that recall of self-relevant health information is susceptible to self-enhancement bias. PMID- 16719616 TI - The effects of applied tension on symptoms in French-speaking blood donors: a randomized trial. AB - Blood-donation-related symptoms such as dizziness and weakness discourage people from participating in this important health-related activity. Four hundred sixty seven young adult, French-speaking blood donors were randomly assigned to (a) a condition in which they learned a possible preventive technique called applied tension and were asked to practice it from the time they got on the donation chair until they were just about to get up, (b) a placebo condition in which they learned applied tension and were asked to practice it from the time they got on the chair until the insertion of the donation needle, or (c) a no-treatment control condition. Donors assigned to the treatment condition reported significantly fewer blood-donation-related symptoms than did donors assigned to the other conditions and rated their likelihood of returning to give blood again as greater than did those in the no treatment condition. Among donors whose chairs were not reclined, participants in the treatment condition had significantly smaller heart rate reactions to blood donation than did those in the other conditions. PMID- 16719617 TI - Remembering the message: the use of a reminder cue to increase condom use following a safer sex intervention. AB - Condom use interventions may be more powerful if they provide cues to recall safe sex messages when sexual activity occurs. The authors tested this notion by assigning sexually active introductory psychology students (N = 196) to a standard safe-sex intervention, a safe-sex with reminder intervention, or a control (drinking and driving) intervention. Participants assigned to the reminder intervention were given a "friendship bracelet" to wear and were instructed to have the bracelet remind them of the intervention. In a follow-up session (5-7 weeks later), they were asked questions pertaining to condom use. Of the 125 participants who had engaged in sexual intercourse, condom use at last intercourse was higher in the bracelet condition (55%) than in the standard (27%) or control (36%) conditions. The authors also found that the bracelet remained effective, even when participants were under the influence of alcohol. These findings therefore imply that health intervention programs may be more efficacious if they include strategies such as reminder cues to increase the salience of health information in the appropriate contexts. PMID- 16719619 TI - Comparing patients' predicted test scores from a regression equation with their obtained scores: a significance test and point estimate of abnormality with accompanying confidence limits. AB - In contrast to the standard use of regression, in which an individual's score on the dependent variable is unknown, neuropsychologists are often interested in comparing a predicted score with a known obtained score. Existing inferential methods use the standard error for a new case (s-subN+1) to provide confidence limits on a predicted score and hence are tailored to the standard usage. However, s-subN+1 can be used to test whether the discrepancy between a patient's predicted and obtained scores was drawn from the distribution of discrepancies in a control population. This method simultaneously provides a point estimate of the percentage of the control population that would exhibit a larger discrepancy. A method for obtaining confidence limits on this percentage is also developed. These methods can be used with existing regression equations and are particularly useful when the sample used to generate a regression equation is modest in size. Monte Carlo simulations confirm the validity of the methods, and computer programs that implement them are described and made available. PMID- 16719620 TI - The role of the corpus callosum in dichotic listening: a combined morphological and diffusion tensor imaging study. AB - The objective of the present study was to examine the role of the corpus callosum (CC) in dichotic listening. For this purpose, 40 right-handed healthy male participants were assessed with morphological and diffusion tensor imaging, which yielded macrostructural (midsagittal area) and microstructural (mean diffusion, fractional anisotropy) measures of the total CC and predefined subregions. Applying the standard consonant-vowel Bergen Dichotic Listening Test (including 3 different attentional instructions), the authors found dichotic listening performance to be substantially related to the integrity of the CC at both macro- and microstructural levels. Results indicate a dual role of the CC; it is relevant not only for the stimulus-driven (bottom-up) transfer of left-ear input to the left hemisphere but also for its attentional (top-down) modulation. PMID- 16719621 TI - Instrumental learning and relearning in individuals with psychopathy and in patients with lesions involving the amygdala or orbitofrontal cortex. AB - Previous work has shown that individuals with psychopathy are impaired on some forms of associative learning, particularly stimulus-reinforcement learning (Blair et al., 2004; Newman & Kosson, 1986). Animal work suggests that the acquisition of stimulus-reinforcement associations requires the amygdala (Baxter & Murray, 2002). Individuals with psychopathy also show impoverished reversal learning (Mitchell, Colledge, Leonard, & Blair, 2002). Reversal learning is supported by the ventrolateral and orbitofrontal cortex (Rolls, 2004). In this paper we present experiments investigating stimulus-reinforcement learning and relearning in patients with lesions of the orbitofrontal cortex or amygdala, and individuals with developmental psychopathy without known trauma. The results are interpreted with reference to current neurocognitive models of stimulus reinforcement learning, relearning, and developmental psychopathy. PMID- 16719622 TI - A novel set-shifting modification of the iowa gambling task: flexible emotion based learning in schizophrenia. AB - Although it might seem that people with schizophrenia would perform poorly on measures of emotion-based learning, several studies have shown normal levels of performance on the Iowa Gambling Task (IGT; C. E. Y. Evans, C. H. Bowman, & O. H. Turnbull, 2005; L. M. Ritter, J. H. Meador-Woodruff, & G. W. Dalack, 2004; B. Shurman, W. P. Horan, & K. H. Nuechterlein, 2005; K. E. Wilder, D. R. Weinberger, & T. E. Goldberg, 1998). The present article describes a newly developed modification of the IGT involving initial familiarization with the basic contingency pattern then 3 periods of contingency shift. Control participants showed substantial gains during the later trials of each shift period. Analyzed in terms of positive symptoms, those with schizophrenia were little different from control participants. Those high in negative symptoms could perform the basic task but showed remarkably poor performances (no better than chance) in the shift phases, retaining a preference for decks that had previously been "good," even when they experienced substantial losses. PMID- 16719623 TI - The influence of dopamine on semantic activation in Parkinson's disease: evidence from a multipriming task. AB - Research has suggested that semantic processing deficits in Parkinson's disease (PD) are related to striatal dopamine deficiency. As an investigation of the influence of dopamine on semantic activation in PD, 7 participants with PD performed a lexical-decision task when on and off levodopa medication. Seven healthy controls matched to the participants with PD in terms of sex, age, and education also participated in the study. By use of a multipriming paradigm, whereby 2 prime words were presented prior to the target word, semantic priming effects were measured across stimulus onset asynchronies (SOAs) of 250 ms and 1,200 ms. The results revealed a similar pattern of priming across SOAs for the control group and the PD participants on medication. In contrast, within-group comparisons revealed that automatic semantic activation was compromised in PD participants when off medication. The implications of these results for the neuromodulatory influence of dopamine on semantic processing in PD are discussed. PMID- 16719624 TI - How does the brain support script comprehension? A study of executive processes and semantic knowledge in dementia. AB - The neuropsychological substrate of scripts, routines which guide much of human behavior, is unclear. We propose a model of script comprehension characterized by the interaction of semantic knowledge for script content, and executive resources that organize this knowledge into goal directed behavior. We examined these neuropsychological components by asking participants with Alzheimer's disease (AD) and frontotemporal dementia (behavioral disorder/dysexecutive syndrome (BDD) and semantic dementia (SD) subtypes), to judge the coherence of four-phrase scripts. The BDD group detected significantly fewer sequencing errors than semantic errors; the AD and SD groups detected these errors with equal frequency. Independent semantic measures predicted both semantic and sequencing script errors, while executive measures predicted sequencing errors only. Findings support a multi-component model of script comprehension. PMID- 16719625 TI - Semantic memory in Alzheimer's disease and the frontotemporal dementias: a longitudinal study of 236 patients. AB - Using semantic dementia (SD) as a reference point, the authors assessed semantic memory in four other neurodegenerative disorders: progressive nonfluent aphasia (PNFA), frontal variant frontotemporal dementia (fvFTD), Alzheimer's disease (AD), and posterior cortical atrophy (PCA). Individuals with SD were more impaired than other groups on semantic measures and showed a characteristic pattern across tasks: category fluency (CF) worse than letter fluency (LF), naming worse than comprehension, and visual and verbal comprehension equally affected, suggesting disruption to an amodal semantic system. Individuals with AD demonstrated a similar pattern to a milder degree. Although PNFA, fvFTD, and PCA groups had abnormal scores (relative to controls) on most semantic measures, their differing patterns across measures indicate that the apparent semantic impairment in these conditions is largely secondary to other factors. PMID- 16719626 TI - Increased interhemispheric interaction is associated with earlier offset of childhood amnesia. AB - Experiment 1 found that mixed-handedness, which is associated with increased interaction between the left and right cerebral hemispheres relative to strong right-handedness, was associated with an earlier offset of childhood amnesia. In Experiment 2, bilateral saccadic eye movements, which have been shown to enhance interhemispheric interaction, were also associated with an earlier offset of childhood amnesia. These results build upon a growing body of research indicating an interhemispheric basis for the retrieval of episodic memories. Moreover, the results of Experiment 2 suggest that interhemispheric interaction has its effect on the retrieval, not encoding, of episodic memories. PMID- 16719627 TI - Exploring the neural organization of person-related knowledge: lateralization of lesion, category specificity, and stimulus modality effects. AB - While it is generally agreed that the right fusiform gyrus is specialized for face recognition, the question of whether knowledge about persons is lateralized in the temporal lobes is more contentious. Does knowledge about people differ from other kinds of object knowledge with respect to brain laterality? Are side of-lesion effects mediated by stimulus modality? This study aimed to investigate these questions by comparing patients with left temporal (LT) (n=8) and right temporal (RT) (n=11) lesions to control subjects (n=12) on verbal and visual tests of people, buildings, and objects. The RT group was impaired at recognizing famous faces, but not at choosing the picture of a famous building or a famous name from nonfamous distracters. The LT group was impaired at naming people, buildings, and objects, regardless of stimulus modality. When presemantic processing was controlled for, neither patient group was impaired in producing person-specific knowledge to faces or names, supporting the notion that semantic knowledge for people as for other kinds of objects, is stored in a distributed network across both hemispheres, regardless of stimulus modality. PMID- 16719628 TI - Risky decision making assessed with the gambling task in adults with HIV. AB - Decision making was assessed using a laboratory gambling task in 67 adults with the Human Immunodeficiency Virus (HIV+) and in 19 HIV-seronegative (HIV-) control participants. Neurocognitive test performance across several domains was also analyzed to examine potential cognitive mechanisms of gambling task performance. As predicted, the HIV+ group performed worse on the gambling task, indicating greater risky decision making. Specifically, the HIV+ group selected more cards from the "risky" or disadvantageous deck that included relatively large payoffs but infrequent large penalties. The control group also selected such risky cards but quickly learned to avoid them. Exploratory analyses also indicated that in the HIV+ group, but not in the control group, gambling task performance was correlated with Stroop Interference performance and long delay free recall on the California Verbal Learning Test, suggesting the role of inhibitory processes and verbal memory in the poorer gambling task performance in HIV. These findings indicate the usefulness of the gambling task as a laboratory tool to examine risky decision making and cognition in the HIV population. PMID- 16719630 TI - Visual selective attention in Parkinson's disease: dissociation of exogenous and endogenous inhibition. AB - Impairment in the inhibitory mechanism of visual selective attention in patients with Parkinson's disease (PD) is controversial. The present study sought to understand disparate findings in a manner analogous to the relative preservation of exogenously evoked movement and impairment of endogenously evoked movement. The authors examined inhibition of return (i.e., exogenously evoked inhibition; IOR) and negative priming (i.e., endogenously evoked inhibition; NP) in a group of 14 patients with PD and 14 healthy controls (HC). Unlike the HC, who demonstrated significant inhibition in both tasks, the group with PD demonstrated intact inhibition only in the IOR task. Dopamine replacement therapy did not affect performance. The findings are discussed within the context of a model that differentiates the essential involvement of the basal ganglia for endogenously evoked spatial inhibition. PMID- 16719629 TI - Focused and shifting attention in children with heavy prenatal alcohol exposure. AB - Attention deficits are a hallmark of the teratogenic effects of alcohol. However, characterization of these deficits remains inconclusive. Children with heavy prenatal alcohol exposure and nonexposed controls were evaluated using a paradigm consisting of three conditions: visual focus, auditory focus, and auditory-visual shift of attention. For the focus conditions, participants responded manually to visual or auditory targets. For the shift condition, participants alternated responses between visual targets and auditory targets. For the visual focus condition, alcohol-exposed children had lower accuracy and slower reaction time for all intertarget intervals (ITIs), while on the auditory focus condition, alcohol-exposed children were less accurate but displayed slower reaction time only on the longest ITI. Finally, for the shift condition, the alcohol-exposed group was accurate but had slowed reaction times. These results indicate that children with heavy prenatal alcohol exposure have pervasive deficits in visual focused attention and deficits in maintaining auditory attention over time. However, no deficits were noted in the ability to disengage and reengage attention when required to shift attention between visual and auditory stimuli, although reaction times to shift were slower. PMID- 16719631 TI - Selective attention deficits and subjective fatigue following traumatic brain injury. AB - The present study aimed to investigate the relationship between subjective fatigue and selective attention deficits following traumatic brain injury (TBI). Forty-six participants with mild-severe TBI and 46 healthy controls completed fatigue scales (Visual Analogue Scale--Fatigue, Fatigue Severity Scale [FSS] and Causes of Fatigue Questionnaire [COF]), and attentional measures including subtests from the Test of Everyday Attention, and the Complex Selective Attention Task (C-SAT). TBI participants reported greater fatigue on the FSS and COF, performed more slowly on attentional measures, and made more errors on the C-SAT. After controlling for anxiety and depression, fatigue was significantly correlated with performance only on the C-SAT. Findings suggest a relationship between subjective fatigue and impairment on tasks requiring higher order attentional processes. PMID- 16719632 TI - Orthopsychiatry in the 21st century. PMID- 16719633 TI - Differences in patterns of symptom attribution in diagnosing schizophrenia between African American and non-African American clinicians. AB - The authors examined clinician race differences in symptom attribution patterns in diagnosing psychiatric inpatients from a low-income, African American community. Different decision models were applied to patients based on clinician race. African American clinicians diagnosed schizophrenia with higher odds than non-African American clinicians when they believed hallucinations were present and avoided that diagnosis with lower odds when they considered substance abuse issues. Non-African American clinicians usually related the attribution of negative symptoms to the diagnosis of schizophrenia while African American clinicians did not make this linkage. The study highlights the need for more detailed examination of cultural influences on diagnostic judgments. PMID- 16719634 TI - Seeking help a second time: parents'/caregivers' characterizations of previous experiences with mental health services for their children and perceptions of barriers to future use. AB - This study examines the relationship between urban parents'/caregivers' previous experiences obtaining mental health care for their children and their perceptions of barriers to their children's use of services in the future. Assessments of prior treatment outcome and aspects of relationships with former providers were linked to endorsements of doubt about the utility of treatment as a potential barrier to the children's use of services in the future and the number of barriers parents endorsed. Implications for urban child mental health service delivery are drawn. PMID- 16719635 TI - Post-9/11 helpseeking by New York City parents on behalf of highly exposed young children. AB - This study examines factors related to helpseeking among New York City parents on behalf of their young children after the September 11th terrorist attacks. Data were gathered from 180 parents about their children (under age 5) through in depth parent interviews 9-12 months postdisaster. Parents were asked to describe their children's disaster-related experiences, their own and their children's mental health status, and post-9/11 helpseeking behavior for their children. Predictors of parental helpseeking for children's services included the emergence of new fears in children since 9/11, parent symptoms of depression, and parents' own helpseeking. The strongest predictor was children's direct exposure to the attacks. Fifteen percent (n = 27) of parents sought services for their very young children. Findings suggest that following 9/11, a familial orientation to helpseeking combined with children's specific disaster-related experiences may provide a basis for seeking services for young children, rather than children's apparent mental health status. PMID- 16719636 TI - Anger, guilt, and intergenerational communication of trauma in the interpersonal narratives of second generation Holocaust survivors. AB - The nature and expression of anger and guilt in sons and daughters of Holocaust survivors were studied by a quantitative and qualitative analysis of relationship narratives. Findings are discussed in relation to the reciprocal overprotectiveness between the surviving parents and their children in the context of intergenerational communication of trauma. PMID- 16719637 TI - Toward a positive psychology of mental retardation. AB - Researchers and practitioners alike have long focused on the external life conditions, adaptive behavior, and inclusion of persons with mental retardation. Using breakthroughs in positive psychology, this article proposes a new research agenda focused on the positive, internal states of those with mental retardation. It shows how major movements in the mental retardation field--quality of life, dual diagnosis, personality motivation, and families--have succeeded in some arenas but failed to address happiness and well-being. Examples of happiness--of positive emotions, flow, strengths, and virtues--are offered in people with genetic causes of mental retardation. Complexities related to etiology, measurement, flow, and a meaningful life are described, as is the vital role that mental retardation can play in the emerging science of positive psychology. PMID- 16719638 TI - Integrating evidence-based practice in the child mental health agency: a template for clinical and organizational change. AB - This article describes a linked set of activities designed to address the present "science-to-service" gap in evidence-based practice from the perspective of the nonprofit child mental health agency: (a) implementing "systems-of-care" principles; (b) logic modeling; (c) strategic partnerships with external researchers; (d) selective, in-depth program evaluation; (e) critical comparison with a limited number of evidence-based practice models of national significance; and (f) internal and external dissemination activities oriented to evidence-based practice. Implications are drawn both for continuous quality improvement of existing clinical services and for the potential for agencies to contribute high quality, science-based information to child mental health services, singly and through the future development of a national network of experimenting child mental health agencies. PMID- 16719639 TI - Ethnicity and life quality of recently discharged inpatients with schizophrenia. AB - Three months after inpatient discharge, quality of life was examined in 259 individuals with schizophrenia. As in findings by Lehman, Rachuba, and Postrado (1995), Black participants reported a higher quality than Whites after controlling for gender, degree of psychopathology, functioning level, substance abuse, and baseline life quality. Implications are discussed. PMID- 16719640 TI - Fifty delinquents in juvenile and adult court. AB - From an analysis of developmental assessments of 50 delinquents, the author found that those who received long adult sentences showed the same disabilities, past trauma, and immaturity as those who remained in the juvenile system. Half of those sent to adult prison had no prior arrests, including three 13-year-olds, a 14-year-old, and two 15-year-olds. A majority of the 50 youth had characteristics of teenagers likely to outgrow delinquency, yet more than half of them were sentenced to adult prison. The author encouraged experts to present specific recommendations for rehabilitative services to address the juveniles' disabilities, recovery from trauma, and development of mature thinking in response to the court's concerns about future reoffending. PMID- 16719641 TI - An empirically supported and culturally specific engagement and intervention strategy for African American adolescent males. AB - The need for effective culturally responsive treatments has become more urgent as the number of ethnic minority clients continues to increase. Previous research with a clinically referred sample of substance-abusing African American inner city teenagers found that treatment engagement increased when cultural content was incorporated in the therapeutic process (Jackson-Gilfort, Liddle, Tejeda, & Dakof, 2001). This article amplifies these findings by offering clinical guidelines for how to develop and implement culturally specific interventions that contribute to the therapeutic engagement of African American adolescent males. Clinical outcomes may be improved by integrating culturally responsive intervention methods within a multisystemic approach to the adolescent's treatment. PMID- 16719642 TI - Campus mental health services: recommendations for change. AB - College officials indicate that the number of students with serious mental illnesses has risen significantly. Recent media attention surrounding several high profile suicides has opened discussion of mental illness on campus. The authors summarize literature on college students and mental illness, including barriers to service receipt. Recommendations to improve campus-based responses to serious mental illness are presented on the basis of well-accepted service principles. PMID- 16719644 TI - Is three a crowd? Clients, clinicians, and managed care. AB - How does therapy change when therapists work under managed care plans? To explore this question, the authors conducted in-depth, semistructured interviews with therapists. The interview texts were analyzed using a structured thematic coding system. A central theme in the interviews was a culture clash between managed care companies and these therapists. Working for managed care organizations demanded several practices that violated the therapists' standard of care and professional ethics. Also, participants reported that managed care personnel misrepresented the nature of psychotherapy to clients, thereby undermining the therapeutic work and the therapist-client relationship. If these therapists' experiences are representative, the growth of managed care has serious implications for the nature of psychotherapy, practitioners' integrity and morale, and the public image of the therapy professions. PMID- 16719643 TI - Risk factors for deliberate self-harm among female college students: the role and interaction of childhood maltreatment, emotional inexpressivity, and affect intensity/reactivity. AB - Despite the clinical importance of deliberate self-harm, research on the risk factors for self-harm among nonclinical populations has been limited. This study examined the role of childhood maltreatment, emotional inexpressivity, and affect intensity/reactivity in the self-harm behavior of 249 female college students. Childhood maltreatment and low positive affect intensity/reactivity reliably distinguished women with frequent self-harm from women with no history of self harm, as did the combination of greater maltreatment, greater inexpressivity, and higher levels of affect intensity/reactivity (global and negative). Among women with a history of self-harm, emotional inexpressivity was associated with more frequent self-harm, as was the combination of greater maltreatment, greater inexpressivity, and lower levels of positive affect intensity/reactivity. PMID- 16719645 TI - Abstinence among female adolescents: do parents matter above and beyond the influence of peers? AB - The purpose of this study was to determine whether parental variables predict abstinence above and beyond the influence of peer variables in a sample of African American adolescent females (n = 568). In order to determine predictors of abstinence, hierarchical logistic regressions were conducted for the entire sample, the younger adolescent girls, and the older adolescent girls. Results indicated that for the total sample and older adolescents, both parental sexual attitudes and parent-adolescent relationship quality predicted abstinence after accounting for the variance associated with peer variables. For the younger girls, perceived parental attitudes were the only significant predictor. These findings suggest that adolescent sexual risk reduction interventions may benefit from including parents, stressing the importance of articulating parental sexual attitudes, and highlighting the significance of the parent-adolescent relationship. PMID- 16719646 TI - Children enrolled in public pre-K: the relation of family life, neighborhood quality, and socioeconomic resources to early competence. AB - This article presents data on the family and social environments of 501 children enrolled in public sponsored pre-K in 5 states and tests the relation of these resources to child competence. Structured interviews and questionnaires provide information from parents about the family's social and economic status. Direct assessments and teacher reports provide data on children's literacy, numeracy, and behavioral problems. A majority of the children served in public pre-K lived in poverty and showed decrements in language but not in other domains. A socioeconomic resource factor consisting of parental education, household income, and material need predicted all domains of children's functioning. Children from households high in socioeconomic resources entered pre-K with more well developed language and math skill but fewer behavioral problems than their disadvantaged peers. Neighborhood quality status was related to language competence and mother's marital status to math competence. Neighborhood quality and income level may have their impact on child competence through their relation to dyadic quality and the health and the psychological well-being of the parents. PMID- 16719647 TI - Instantiated features and the use of "rules.". AB - Classification "rules" in expert and everyday discourse are usually deficient by formal standards, lacking explicit decision procedures and precise terms. The authors argue that a central function of such weak rules is to focus on perceptual learning rather than to provide definitions. In 5 experiments, transfer following learning of family resemblance categories was influenced more by familiar-appearing features than by novel-appearing features equally acceptable under the rule. This occurred both when rules were induced and when rules were given at the beginning of instruction. To model this and other phenomena in categorization, features must be represented on 2 levels: informational and instantiated. These 2 feature levels are crucial to provide broad generalization while reflecting the known peculiarities of a complex world. PMID- 16719648 TI - Predicting the near and distant future. AB - Four studies investigated individuals' confidence in predicting near future and distant future outcomes. Study 1 found that participants were more confident in theory-based predictions of psychological experiments when these experiments were expected to take place in the more distant future. Studies 2-4 examined participants' confidence in predicting their performance on near and distant future tests. These studies found that in predicting their more distant future performance, participants disregarded the format of the questions (e.g., multiple choice vs. open ended) and relied, instead, on their perceived general knowledge (e.g., history knowledge). Together, the present studies demonstrate that predictions of the more distant future are based on relatively abstract information. Individuals feel more confident in predicting the distant future than the near future when the predictions concern outcomes that are implied by relatively abstract information. PMID- 16719649 TI - Insight and strategy in multiple-cue learning. AB - In multiple-cue learning (also known as probabilistic category learning) people acquire information about cue-outcome relations and combine these into predictions or judgments. Previous researchers claimed that people can achieve high levels of performance without explicit knowledge of the task structure or insight into their own judgment policies. It has also been argued that people use a variety of suboptimal strategies to solve such tasks. In three experiments the authors reexamined these conclusions by introducing novel measures of task knowledge and self-insight and using "rolling regression" methods to analyze individual learning. Participants successfully learned a four-cue probabilistic environment and showed accurate knowledge of both the task structure and their own judgment processes. Learning analyses suggested that the apparent use of suboptimal strategies emerges from the incremental tracking of statistical contingencies in the environment. PMID- 16719650 TI - From recurrent choice to skill learning: a reinforcement-learning model. AB - The authors propose a reinforcement-learning mechanism as a model for recurrent choice and extend it to account for skill learning. The model was inspired by recent research in neurophysiological studies of the basal ganglia and provides an integrated explanation of recurrent choice behavior and skill learning. The behavior includes effects of differential probabilities, magnitudes, variabilities, and delay of reinforcement. The model can also produce the violation of independence, preference reversals, and the goal gradient of reinforcement in maze learning. An experiment was conducted to study learning of action sequences in a multistep task. The fit of the model to the data demonstrated its ability to account for complex skill learning. The advantages of incorporating the mechanism into a larger cognitive architecture are discussed. PMID- 16719651 TI - SSL: a theory of how people learn to select strategies. AB - The assumption that people possess a repertoire of strategies to solve the inference problems they face has been raised repeatedly. However, a computational model specifying how people select strategies from their repertoire is still lacking. The proposed strategy selection learning (SSL) theory predicts a strategy selection process on the basis of reinforcement learning. The theory assumes that individuals develop subjective expectations for the strategies they have and select strategies proportional to their expectations, which are then updated on the basis of subsequent experience. The learning assumption was supported in 4 experimental studies. Participants substantially improved their inferences through feedback. In all 4 studies, the best-performing strategy from the participants' repertoires most accurately predicted the inferences after sufficient learning opportunities. When testing SSL against 3 models representing extensions of SSL and against an exemplar model assuming a memory-based inference process, the authors found that SSL predicted the inferences most accurately. PMID- 16719652 TI - Tradeoffs and theory: the double-mediation model. AB - Most theories of decision making suggest that, when options imply tradeoffs between their attributes, conflict increases as tradeoff size increases, because greater sacrifices are to be incurred in choosing one option instead of another. An alternative view is that conflict decreases as tradeoff size increases, because stronger arguments can be made for any decision. The authors propose a unified model, the double-mediation model, which combines the mediating effects of sacrifice and argumentation. Our model generally predicts an inverse U-shaped relation between tradeoff size and conflict. Results support this prediction. Also, when the decision situation increases the mediating effect of sacrifice relative to that of argumentation, the relation between tradeoff size and conflict changes in an upward direction; conversely, when the decision situation increases the mediating effect of argumentation relative to that of sacrifice, the relation changes in a downward direction. Results support these predictions as well. Commonalities and differences between our model and other formulations are discussed. PMID- 16719653 TI - The influence of improper sets of information on judgment: how irrelevant information can bias judged probability. AB - This article introduces 2 new sources of bias in probability judgment, discrimination failure and inhibition failure, which are conceptualized as arising from an interaction between error prone memory processes and a support theory like comparison process. Both sources of bias stem from the influence of irrelevant information on participants' probability judgments, but they postulate different mechanisms for how irrelevant information affects judgment. The authors used an adaptation of the proactive interference (PI) and release from PI paradigm to test the effect of irrelevant information on judgment. The results of 2 experiments support the discrimination failure account of the effect of PI on probability judgment. In addition, the authors show that 2 commonly used measures of judgment accuracy, absolute and relative accuracy, can be dissociated. The results have broad implications for theories of judgment. PMID- 16719654 TI - Dissociating uncertainty responses and reinforcement signals in the comparative study of uncertainty monitoring. AB - Although researchers are exploring animals' capacity for monitoring their states of uncertainty, the use of some paradigms allows the criticism that animals map avoidance responses to error-causing stimuli not because of uncertainty monitored but because of feedback signals and stimulus aversion. The authors addressed this criticism with an uncertainty-monitoring task in which participants completed blocks of trials with feedback deferred so that they could not associate reinforcement signals to particular stimuli or stimulus-response pairs. Humans and 1 of 2 monkeys were able to make cognitive, decisional uncertainty responses that were independent of feedback or reinforcement history within a task. This finding unifies the comparative literature on uncertainty monitoring. The dissociation of performance from reinforcement has theoretical implications, and the deferred-feedback technique has many applications. PMID- 16719655 TI - Is the binding of visual features in working memory resource-demanding? AB - The episodic buffer component of working memory is assumed to play a role in the binding of features into chunks. A series of experiments compared memory for arrays of colors or shapes with memory for bound combinations of these features. Demanding concurrent verbal tasks were used to investigate the role of general attentional processes, producing load effects that were no greater on memory for feature combinations than for the features themselves. However, the binding condition was significantly less accurate with sequential rather than simultaneous presentation, especially for items earlier in the sequence. The findings are interpreted as evidence of a relatively automatic but fragile visual feature binding mechanism in working memory. Implications for the concept of an episodic buffer are discussed. PMID- 16719656 TI - How to hit Scylla without avoiding Charybdis: comment on Perruchet, Tyler, Galland, and Peereman (2004). AB - M. Pena, L. L. Bonatti, M. Nespor, and J. Mehler argued that humans compute nonadjacent statistical relations among syllables in a continuous artificial speech stream to extract words, but they use other computations to determine the structural properties of words. Instead, when participants are familiarized with a segmented stream, structural generalizations about words are quickly established. P. Perruchet, M. D. Tyler, N. Galland, and R. Peereman criticized M. Pena et al.'s work and dismissed their results. In this article, the authors show that P. Perruchet et al.'s criticisms are groundless. PMID- 16719658 TI - Time as a guide to cause. AB - How do people learn causal structure? In 2 studies, the authors investigated the interplay between temporal-order, intervention, and covariational cues. In Study 1, temporal order overrode covariation information, leading to spurious causal inferences when the temporal cues were misleading. In Study 2, both temporal order and intervention contributed to accurate causal inference well beyond that achievable through covariational data alone. Together, the studies show that people use both temporal-order and interventional cues to infer causal structure and that these cues dominate the available statistical information. A hypothesis driven account of learning is endorsed, whereby people use cues such as temporal order to generate initial models and then test these models against the incoming covariational data. PMID- 16719659 TI - Context switch effects on acquisition and extinction in human predictive learning. AB - Four experiments tested context switch effects on acquisition and extinction in human predictive learning. A context switch impaired probability judgments about a cue-outcome relationship when the cue was trained in a context in which a different cue underwent extinction. The context switch also impaired judgments about a cue trained in a context different from the extinction context, whenever this training was concurrent with extinction of another cue. After extinction, new cue-outcome relationships learned, even in a different task, became context specific. Moreover, renewal was consistently observed. It is suggested that context switch effects result from a process by which ambiguity leads participants to attend to the contexts. PMID- 16719660 TI - Qualitative differences between implicit and explicit sequence learning. AB - Four experiments investigate the differences between implicit and explicit sequence learning concerning their resilience to structural and superficial task changes. A superficial change that embedded the SRT task in the context of a selection task, while maintaining the sequence, did selectively hinder the expression of implicit learning. In contrast, a manipulation that maintained the task surface, but decreased the sequence validity, affected the expression of learning specifically when it was explicit. These results are discussed in the context of a dynamic framework (Cleeremans & Jimenez, 2002), which assumes that implicit knowledge is specially affected by contextual factors and that, as knowledge becomes explicit, it allows for the development of relevant metaknowledge that modulates the expression of explicit knowledge. PMID- 16719661 TI - The intention superiority effect in motor skill learning. AB - Three experiments were conducted to determine if the intention to perform motor sequences in the future results in similar patterns of activation and inhibition as observed for verbal scripts. In Experiments 1 and 2, intention was induced by informing one group that they would be tested on the tasks following acquisition; the other group was not informed of the retention test. Recognition tests administered prior to and after the retention test indicated a strong intention superiority effect. However, intention instructions provided either at the end of acquisition (Experiment 1) or before acquisition (Experiment 2) failed to impact acquisition or retention performance of the motor sequences, but did influence the latency of responding on the retention test. Experiment 3 was designed to replicate the results of Experiments 1 and 2 using a within-subjects design and extend these findings to observation. The results indicated that intention instructions resulted in a strong intention superiority effect for both the physical and observational practice participants, but the performance on the intentional tasks was enhanced only for the observational practice group. PMID- 16719662 TI - Individual skill differences and large-scale environmental learning. AB - Spatial skills are known to vary widely among normal individuals. This project was designed to address whether these individual differences are differentially related to large-scale environmental learning from route (ground-level) and survey (aerial) perspectives. Participants learned two virtual environments (route and survey) with limited exposure and tested on judgments about relative locations of objects. They also performed a series of spatial and nonspatial component skill tests. With limited learning, performance after route encoding was worse than performance after survey encoding. Furthermore, performance after route and survey encoding appeared to be preferentially linked to perspective and object-based transformations, respectively. Together, the results provide clues to how different skills might be engaged by different individuals for the same goal of learning a large-scale environment. PMID- 16719663 TI - Allocentric-heading recall and its relation to self-reported sense-of-direction. AB - A sense of direction (SOD) computes the body's facing direction relative to a reference frame grounded in the environment. The authors report on three experiments in which they used a heading-recall task to tap the functioning of a SOD system and then correlated task performance with self-reported SOD as a convergent test of the task's construct validity. On each heading-recall trial, the participant judged the photographer's allocentric heading when photographing a pictured outdoor scene. Participants were tested over the full range of SOD ratings in Experiment 1, and in Experiments 2 and 3 heading-recall at the SOD extremes was tested. In all experiments, there was wide variability in heading recall accuracy that covaried with self-rated SOD. Parametric manipulation of various task parameters revealed some likely functional properties of the SOD system. The results support the psychological reality of a SOD system and further indicate that there are large individual differences in the efficacy with which the system functions. PMID- 16719664 TI - Specificity effects in training and transfer of speeded responses. AB - In 3 experiments, participants, on signal, moved a cursor from a central position to 1 of 8 numerically labeled locations on the circumference of a clock face. Movements were controlled by a mouse in 1 of 4 conditions: vertical reversal, horizontal reversal, combined reversals, or normal (i.e., no reversals). Participants were trained in 1, 2, or 3 of these conditions and were tested 1 week later with either the same or a different condition. There were improvements across training and perfect retention across the delay. There was little or no transfer, however, even when training involved combined reversals or multiple conditions. These results illustrate severe specificity of training and are interpreted in terms of acquired inhibition of normal responses. PMID- 16719665 TI - Hypothetical and certain inferences from conditional arguments read in texts. AB - In 3 experiments, affirmative and hypothetical probes were presented after narrative texts containing conditional arguments. According to the data, readers represented modus ponens deductions as certain, except when it was only a weakly necessary cause of a given effect. They represented any logically invalid inferences resulting from affirming the conditional consequent as hypothetical, except when it was the effect of strongly sufficient cause. Accordingly, readers must be processing conditional syntax as an asymmetric constraint. However, the underlying causal knowledge can be sufficient either to discredit or warrant the inferences. Thus according to the theory of natural logic, readers can draw formal deductions and be convinced of their necessity. This provides further evidence that readers can represent their inferences as hypothetical (N. Campion, 2004). PMID- 16719666 TI - The negated conditional: a litmus test for the suppositional conditional? AB - Under the suppositional account of conditionals, when people think about a conditional assertion, "if p then q," they engage in a mental simulation in which they imagine p holds and evaluate the probability that q holds under this supposition. One implication of this account is that belief in a conditional equates to conditional probability [P(q/p)]. In this paper, the authors examine a further implication of this analysis with respect to the wide-scope negation of conditional assertions, "it is not the case that if p then q." Under the suppositional account, nothing categorically follows from the negation of a conditional, other than a second conditional, "if p then not-q." In contrast, according to the mental model theory, a negated conditional is consistent only with the determinate state of affairs, p and not-q. In 4 experiments, the authors compare the contrasting predictions that arise from each of these accounts. The findings are consistent with the suppositional theory but are incongruent with the mental model theory of conditionals. PMID- 16719667 TI - All updateable objects in working memory are updated whenever any of them are modified: evidence from the memory updating paradigm. AB - In a series of experiments, participants were required to keep track of 1 or 2 working memory (WM) objects, having to update their values in 80% of the trials. Updating cost, defined as the difference between update and non-update trials, was larger when 2 objects were involved compared with when there was only 1 object was involved. This finding was interpreted as evidence that the updating process encompasses both objects in WM, even though only 1 of them is actually updated. This feature of WM updating is limited to objects defined as "updateable," throughout the trial sequence. The results are explained by the need to reprogram the phonological loop when updating or the need for desynchronization followed by resynchronization of WM contents. PMID- 16719668 TI - The distinctiveness of the word-length effect. AB - The authors report 2 experiments that compare the serial recall of pure lists of long words, pure lists of short words, and lists of long or short words containing just a single isolated word of a different length. In both experiments for pure lists, there was a substantial recall advantage for short words; the isolated words were recalled better than other words in the same list, and there was a reverse word-length effect: Isolated long words were recalled better than isolated short words. These results contradict models that seek to explain the word-length effect in terms of list-based accounts of rehearsal speed or in terms of item-based effects (such as difficulty of assembling items). PMID- 16719669 TI - Exploring a mnemonic debiasing account of the underconfidence-with-practice effect. AB - Judgments of learning (JOLs) underestimate the increase in recall that occurs with repeated study (the underconfidence-with-practice effect; UWP). The authors explore an account in terms of a foresight bias in which JOLs are inflated when the to-be-recalled target highlights aspects of the cue that are not transparent when the cue appears alone and the tendency of practice to alleviate bias by providing learners with cues pertinent to recall. In 3 experiments the UWP effect was strongest for items that induce a foresight bias, but delaying JOLs reduced the debiasing effects of practice, thereby moderating the UWP effect. This occurred when delayed JOLs were prompted by the cue alone (like during testing), not when prompted by the cue-target pair (like during study). PMID- 16719670 TI - Study efficacy and the region of proximal learning framework. AB - One of the most important reasons to investigate human metacognition is its role in directing how people study. However, limited evidence exists that metacognitively guided study benefits learning. Three experiments are presented that provide evidence for this link. In Experiment 1, participants' learning was enhanced when they were allowed to control what they studied. Experiments 2a-d replicated this finding and showed contributions of self-regulated study to learning. Experiments 3a and 3b showed that, when forced to choose among items they did not know, participants chose the easiest items and benefited from doing so, providing evidence for the link between metacognitive monitoring/control and learning, and supporting the region of proximal learning model of study-time allocation. PMID- 16719671 TI - The source of adult age differences in event-based prospective memory: a multinomial modeling approach. AB - Event-based prospective memory involves remembering to perform an action in response to a particular future event. Normal younger and older adults performed event-based prospective memory tasks in 2 experiments. The authors applied a formal multinomial processing tree model of prospective memory (Smith & Bayen, 2004) to disentangle age differences in the prospective component (remembering that you have to do something) and the retrospective component (remembering when to perform the action) of prospective memory performance. The modeling results, as well as more traditional analyses, indicate age differences in the resource demanding prospective component. PMID- 16719672 TI - Hidden covariation detection produces faster, not slower, social judgments. AB - In P. Lewicki's (1986b) demonstration of hidden covariation detection (HCD), responses of participants were slower to faces that corresponded with a covariation encountered previously than to faces with novel covariations. This slowing contrasts with the typical finding that priming leads to faster responding and suggests that HCD is a unique type of implicit process. The authors extended Lewicki's methodology and showed that participants exposed to nonsalient covariations between hair length and personality were subsequently faster to respond to faces with those covariations than to faces without, despite lack of awareness of the critical covariations. This result confirms that people can detect subtle relationships between features of stimuli and that, as with other types of implicit cognition, this detection facilitates responding. PMID- 16719673 TI - Evidence-based practice in psychology. AB - The evidence-based practice movement has become an important feature of health care systems and health care policy. Within this context, the APA 2005 Presidential Task Force on Evidence-Based Practice defines and discusses evidence based practice in psychology (EBPP). In an integration of science and practice, the Task Force's report describes psychology's fundamental commitment to sophisticated EBPP and takes into account the full range of evidence psychologists and policymakers must consider. Research, clinical expertise, and patient characteristics are all supported as relevant to good outcomes. EBPP promotes effective psychological practice and enhances public health by applying empirically supported principles of psychological assessment, case formulation, therapeutic relationship, and intervention. The report provides a rationale for and expanded discussion of the EBPP policy statement that was developed by the Task Force and adopted as association policy by the APA Council of Representatives in August 2005. PMID- 16719674 TI - Testimony and interrogation of minors: assumptions about maturity and morality. AB - This article examines the legal histories and social contexts of testimony and interrogation involving minors, developmental research on suggestibility and judgment, interactions between development and legal/sociological contexts, and the reasoning behind how minors are treated in different legal contexts. The authors argue (a) that young witnesses, victims, and suspects alike possess youthful characteristics that influence their ability to validly inform legal processes, some of which were recently recognized by the Supreme Court as they apply to the juvenile death penalty, and (b) that consideration should be given to reforming current practices in the context of juvenile interrogation. ((c) 2006 APA, all rights reserved). PMID- 16719675 TI - Beyond the hedonic treadmill: revising the adaptation theory of well-being. AB - According to the hedonic treadmill model, good and bad events temporarily affect happiness, but people quickly adapt back to hedonic neutrality. The theory, which has gained widespread acceptance in recent years, implies that individual and societal efforts to increase happiness are doomed to failure. The recent empirical work outlined here indicates that 5 important revisions to the treadmill model are needed. First, individuals' set points are not hedonically neutral. Second, people have different set points, which are partly dependent on their temperaments. Third, a single person may have multiple happiness set points: Different components of well-being such as pleasant emotions, unpleasant emotions, and life satisfaction can move in different directions. Fourth, and perhaps most important, well-being set points can change under some conditions. Finally, individuals differ in their adaptation to events, with some individuals changing their set point and others not changing in reaction to some external event. These revisions offer hope for psychologists and policy-makers who aim to decrease human misery and increase happiness. PMID- 16719676 TI - Venturing a 30-year longitudinal study. AB - Longitudinal inquiry has long been recognized as a uniquely powerful method for seeking understanding of psychological development. A 30-year longitudinal venture is described--its theoretical motivation, methodological rationale, and details of implementation. Some of the novel and implicative findings the study has generated are briefly described. Common to all of the results is an absolute reliance on long-term, widely ranging, independent data. Although specific aspects of the study have appeared over the years, its intentions and scope are recounted only here. By and large, the organizing constructs of ego-control and ego-resiliency find impressive support in various empirical inquiries, here quickly described. Methodologically, a number of savvy research procedures useful and perhaps even necessary in longitudinal research are conveyed. The troublesome burdens but ever-alluring attractions of longitudinal inquiry are noted. A forthcoming Web site will contain the extensive 30-year longitudinal data bank together with explanatory information. Psychological investigators may find these imminently available data resources useful. PMID- 16719679 TI - Comment: taking the theorizing in personality theories seriously. PMID- 16719681 TI - Comment: positive psychology versus the medical model? PMID- 16719682 TI - Using positive psychology with special mental health populations. PMID- 16719683 TI - Comment: treating "the poor"--classism or a rigid loyalty to theory? PMID- 16719684 TI - Comment: classism is overrated. PMID- 16719685 TI - Comment: classism is much more complex. PMID- 16719687 TI - Inactivation of sodium channel Scn8A (Na-sub(v)1.6) in Purkinje neurons impairs learning in Morris water maze and delay but not trace eyeblink classical conditioning. AB - To examine the isolated effects of altered currents in cerebellar Purkinje neurons, the authors used Scn8a-super(flox/flox), Purkinje cell protein-CRE (Pcp CRE) mice in which Exon 1 of Scn8a is deleted only in Purkinje neurons. Twenty male Purkinje Scn8a knockout (PKJ Scn8a KO) mice and 20 male littermates were tested on the Morris water maze (MWM). Subsequently, half were tested in 500-ms delay and half were tested in 500-ms trace eyeblink conditioning. PKJ Scn8a KO mice were impaired in delay conditioning and MWM but not in trace conditioning. These results provide additional support for the necessary participation of cerebellar cortex in normal acquisition of delay eyeblink conditioning and MWM and raise questions about the role, if any, of cerebellar cortex in trace eyeblink conditioning. PMID- 16719688 TI - AMPA receptor GluR2, but not GluR1, subunit deletion impairs emotional response conditioning in mice. AB - Deletions of gria1 or gria2 genes encoding alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic-acid-receptor subunits differ in their effects on appetitive conditioning. The authors investigated whether similar differences would occur in an aversive conditioning test. The ability of a discrete stimulus paired with footshock to subsequently inhibit food-maintained operant responding (conditioned emotional response) was examined in mice with deletions of gria1 or gria2 genes. Whereas gria1 knockout (KO) mice performed normally compared with wild-type (WT) controls, gria2 KO mice displayed no reduction in response rates when the shock-paired stimulus was presented. Nevertheless, gria2 KOs displayed evidence of freezing in a footshock-paired context, indicating that aversive learning could occur. In addition, gria1 KO mice showed some evidence of increased anxiety, and gria2 KOs showed reduced anxiety, in the elevated plus maze. PMID- 16719689 TI - Anxiogenic-like effect of chronic corticosterone in the light-dark emergence task in mice. AB - Chronic hypercortisolemia is a hallmark of neuroendocrine and psychiatric disorders, such as Cushing's disease and depression. Whether cortisol directly contributes to the altered mood and anxiety symptoms seen in these diseases remains unclear. To address this, the authors have modeled hypercortisolemia by administering corticosterone in the drinking water of female Swiss Webster mice for 17 or 18 days (13 mg/kg). Light-dark emergence, startle habituation, and startle reactivity were measured. Chronic but not acute treatment with corticosterone increased the latency to emerge into the light compartment, an anxiogenic-like effect. Chronic corticosterone treatment did not affect startle habituation, but did reduce startle reactivity. This study suggests that chronic hypercortisolemia may contribute to anxiety-related behavior in patients with Cushing's disease and depression. PMID- 16719690 TI - CS-US preexposure effects on trace eyeblink conditioning in young rats: potential implications for functional brain development. AB - Recent studies of delay eyeblink conditioning (EBC) in young rats have demonstrated different effects of various conditioned and unconditioned stimulus (CS-US) preexposure conditions on learning at different ages. The present study extends this research to trace EBC. Subjects experienced 1 of 3 preexposure conditions (paired CS-US, unpaired CS-US, or no stimuli) at either 20 or 24 days of age. Four days later, they were conditioned using either trace (Experiment 1) or delay (Experiment 2) EBC parameters. Results were similar at both ages tested. Paired preexposure facilitated acquisition of delay but not trace relative to context preexposure. Unpaired preexposure impaired acquisition of both delay and trace. These behavioral findings provide a foundation for hypotheses about the functional maturation of cerebellar, hippocampal, and entorhinal learning circuits. PMID- 16719691 TI - Reinforcing properties of ethanol in neonatal rats: involvement of the opioid system. AB - Toward understanding why infant rats ingest high levels of ethanol without initiation procedures, the authors tested effects of mu and kappa receptor antagonists on ethanol reinforcement in neonatal rats. After an intracisternal injection of CTOP (micro antagonist), nor-Binaltorphimine (kappa antagonist), or saline, newborn (3-hr-old) rats were given conditioning pairings of an odor with intraorally infused ethanol or a surrogate nipple with ethanol administered intraperitoneally (to minimize ethanol's gustatory attributes). In each case, these opioid antagonists reduced or eliminated ethanol's reinforcement effect. The same effects occurred with saccharin as the reinforcer in olfactory conditioning. The results imply that activation of mu and kappa receptors, apparently acting jointly, is necessary for reinforcement or that antagonists of this activity impair basic conditioning. PMID- 16719692 TI - Rats bred for high versus low anxiety responses neonatally demonstrate increases in lordosis, pacing behavior, and midbrain 3 alpha, 5 alpha-THP levels as adults. AB - Duration and intensity of lordosis is mediated by actions of the progesterone (P) metabolite, 3 alpha-hydroxy-5 alpha-pregnan-20-one (3 alpha,5 alpha-THP) at GABA sub(A) receptors in the midbrain ventral tegmental area. Because rats selectively bred for infantile vocalizations may differ in endogenous 3 alpha,5 alpha-THP secretion, their sexual behavior, midbrain, and plasma 3 alpha,5 alpha-THP levels as adult rats in behavioral estrus was examined. Rats bred for high rates of infantile vocalizations had shorter latencies and intervals between intromissions and ejaculation, higher lordosis quotients and ratings, more pacing of their sexual contacts, and had higher P and 3 alpha,5 alpha-THP levels in plasma and midbrain than did rats bred for low rats of infantile vocalizations. Thus, levels of 3 alpha,5 alpha-THP in the midbrain are associated with differences in sexual behavior of these rats. PMID- 16719693 TI - Not all rat strains are equal: differential unconditioned fear responses to the synthetic fox odor 2,4,5-trimethylthiazoline in three outbred rat strains. AB - Predator odors induce unconditioned fear in rats; however, the synthetic predator odor 2,4,5-trimethylthiazoline (TMT) either elicits robust fear behavior (e.g., freezing) or no fear responses at all. The authors investigated whether this is due to the use of different outbred rat strains. TMT induced robust freezing in Sprague-Dawley and Long-Evans rats but not in Wistar rats. All 3 strains avoided TMT, but Wistar rats were less sensitive to TMT. Wistar rats are capable of freezing; all 3 strains displayed the same amount of odor-cue conditioned freezing. Thus, TMT is a robust unconditioned fear stimulus in rats, and prior negative results from other laboratories were due to the choice of a rat strain (Wistar) that is less responsive to TMT. PMID- 16719694 TI - Selective lesions of the nucleus basalis magnocellularis impair cognitive flexibility. AB - The authors tested the hypothesis that the cholinergic nucleus basalis magnocellularis (NBM) is involved in solving problems requiring cognitive flexibility. Rats with 192 IgG-saporin lesions of the NBM were assessed for perseveration (i.e., cognitive inflexibility) in the serial reversal of an operant discrimination and during subsequent extinction testing. It was hypothesized that the NBM lesion and control groups would not differ in the acquisition of the initial, simple discrimination, because this task does not demand cognitive flexibility. In contrast, it was hypothesized that the NBM lesion group would show perseveration during serial reversal and extinction testing. Results generally supported these hypotheses, suggesting that the NBM plays an important role in mediating cognitive flexibility. PMID- 16719695 TI - Effect of nucleus basalis magnocellularis cholinergic lesions on fear-like and anxiety-like behavior. AB - Previous research has suggested that cholinergic neurons in the nucleus basalis magnocellularis and substantia innominata (NBM/SI) may be important in mediating aversive states. The authors investigated the effect of NBM/SI cholinergic lesions, induced with 192 IgG saporin, on behavioral measures of aversive states in rats. Behavior in the elevated plus maze and behavioral suppression induced by 2 fear-conditioned stimuli, a tone and a light, were evaluated. Lesions had no effect on any measures in the elevated plus maze but attenuated operant suppression induced by the light and attenuated freezing induced by the tone, though this last effect was not statistically significant. The results of the study suggest that NBM/SI cholinergic neurons may be important in mediating selective aspects of aversive states. PMID- 16719696 TI - Opioid receptors in the midbrain periaqueductal gray regulate prediction errors during pavlovian fear conditioning. AB - The authors used a within-subject blocking design to study the role of ventrolateral periaqueductal gray (v1PAG) opioid receptors in regulating prediction errors during Pavlovian fear conditioning. In Stage I, the authors trained rats to fear conditioned stimulus (CS) A by pairing it with shock. In Stage II, CSA and CSB were co-presented and followed with shock. Two novel stimuli, CSC and CSD, were also co-presented and followed with shock in Stage II. CSA blocked fear from accruing to CSB. Blocking was prevented by systemic pretreatment with naloxone. Blocking was also prevented in a dose-dependent and neuroanatomically specific fashion by vlPAG infusions of the micro-opioid receptor antagonist CTAP. These experiments show that v1PAG micro-opioid receptors contribute to Pavlovian fear learning by regulating predictive error. PMID- 16719697 TI - Effects of bed nucleus of the stria terminalis lesions on conditioned anxiety: aversive conditioning with long-duration conditional stimuli and reinstatement of extinguished fear. AB - Four experiments investigated the effects of lesions of the bed nucleus of the stria terminalis (BNST) on conditioned fear and anxiety. Though BNST lesions did not disrupt fear conditioning with a short-duration conditional stimulus (CS; Experiments 1 and 3), the lesion attenuated conditioning with a longer duration CS (Experiments 1 and 2). Experiment 3 found that lesions attenuated reinstatement of extinguished fear, which relies on contextual conditioning. Experiment 4 confirmed that the lesion reduced unconditioned anxiety in an elevated zero maze. The authors suggest that long-duration CSs, whether explicit cues or contexts, evoke anxiety conditioned responses, which are dissociable from fear responses to shorter CSs. Results are consistent with behavioral and anatomical distinctions between fear and anxiety and with a behavior-systems view of defensive conditioning. PMID- 16719698 TI - Neurophysiological theory of kamin blocking in fear conditioning. AB - Kamin blocking in fear conditioning is thought to reflect diminished processing of the unconditional stimulus (US) in the presence of a conditional stimulus (CS super(+)) that was previously paired with this US. According to Fanselow's (1998) hypothesis, the CS-super(+) drives output from the amygdala that ultimately produces analgesia by causing opiate release onto afferent pain circuits. This hypothesis was explored quantitatively through neurophysiological simulations. The results suggest that opiate-mediated, negative-feedback control of US processing is too slow for efficient blocking of cue conditioning. The reason is that conditioning-produced synaptic modifications can be induced before the opiate-mediated inhibition has any substantial effect on US processing. The results suggest the existence of an additional, faster-acting, inhibitory neurotransmitter in the blocking circuit. PMID- 16719699 TI - Classical tone-shock conditioning induces Zenk expression in the pigeon (Columba livia) hippocampus. AB - The hippocampus is involved in fear conditioning, although the molecular events underlying this function are still under investigation. The authors analyzed the expression of the Zenk proto-oncogene product within the pigeon (Columba livia) hippocampus after training with a classical aversive conditioning protocol using tone-shock associations. Control groups were trained with shock or tone alone or were only exposed to the experimental chamber and manipulated. Experimental pigeons showed significant increases of Zenk expression in the ventromedial region of the hippocampus, whereas both the experimental and shock groups had increased Zenk expression in the dorsal region. The expression of Zenk in specific neuronal populations within the pigeon hippocampus may be indicative of plasticity-associated aversive classical conditioning. PMID- 16719700 TI - Role of instrumental learning in tolerance to cathinone hypophagia. AB - The effects of dl-cathinone on milk intake and motor activity were investigated in bottle- and cannula-fed rats. Acute injections of cathinone produced dose dependent increases in activity in both groups but only produced decreased intake in bottle-fed rats. With chronic injections, tolerance to the suppression of intake developed in the bottle-fed group, accompanied by decreased activity. After the tolerance phase, switching from bottle to cannula feeding produced further increases in intake, whereas switching from cannula to bottle feeding produced decreased intakes. These results suggest that (a) cathinone suppresses intake by inducing locomotion and stereotypy, which interfere with the appetitive phase of feeding, and (b) tolerance to drug-induced hypophagia involves learning to suppress such movements, as proposed by the instrumental learning model. PMID- 16719701 TI - Administration of corticosterone after the first downshift trial enhances consummatory successive negative contrast. AB - Rats given access to a 32% sucrose solution and then downshifted to a 4% solution exhibit less contact with the sipper tube than unshifted controls always given access to 4% solution. This phenomenon, called consummatory successive negative contrast, was facilitated in Experiment 1 by a post-trial injection of corticosterone (3 mg/kg) administered immediately after the first downshift trial. Experiment 2 demonstrated that this facilitatory effect of post-trial corticosterone does not occur when administered 3 hr after the first downshift trial. These results support the hypothesis that corticosterone strengthens an aversive emotional component elicited by the surprising downshift in reward magnitude during the initial downshift trial. PMID- 16719702 TI - Context-dependent modulation by D(1) receptors: differential effects in hippocampus and striatum. AB - Place-specific firing by hippocampal and striatal neurons was recorded simultaneously following injection of a D(1) receptor antagonist (SCH23390) and during spatial working memory task performance. SCH23390-induced changes in unit responses were observed during light and dark test conditions. Although hippocampal place field locations were altered by the contextual change, the reliability and specificity of place fields was disrupted only by combining D(1) antagonism and a change in context. Striatal place field locations were reorganized after either contextual change or D(1) antagonism, without altering place field reliability and specificity. Disrupted velocity encoding by place cells in both regions was induced by darkness, whereas greater stability in acceleration encoding followed removal of D(1) receptor activity. Dopamine may differentially regulate hippocampal context learning and striatum-based predictive codes. PMID- 16719703 TI - Dopamine D(2) receptor activation modulates perceived odor intensity. AB - Dopaminergic modulation affects odor detection thresholds and olfactory discrimination capabilities in rats. The authors show that dopamine D(2) receptor modulation affects odor discrimination capabilities in a manner similar to the modulation of stimulus intensity. Performance in a simultaneous odor discrimination task was systematically altered by manipulations of both odorant concentration and D(2) receptor activation (agonist quinpirole, 0.025-0.5 mg/kg; antagonist spiperone, 0.5 mg/kg). Rats' discrimination performance systematically improved at higher odor concentrations. Blockade of D(2) receptors improved performance equivalent to increasing odor concentration by 2 log units, whereas activation of D(2) receptors reduced odor discrimination performance in a dose dependent manner. Bulbar dopamine release may serve a gain control function in the olfactory system, optimizing its sensitivity to changes in the chemosensory environment. PMID- 16719704 TI - Dorsal hippocampus inhibition disrupts acquisition and expression, but not consolidation, of cocaine conditioned place preference. AB - Cocaine abusers may experience drug craving upon exposure to environmental contexts where cocaine was experienced. The dorsal hippocampus (DHC) is important for contextual conditioning, therefore the authors examined the specific role of the DHC in cocaine conditioned place preference (CPP). Muscimol was used to temporarily inhibit the DHC and was infused before conditioning sessions or tests for CPP to investigate acquisition and expression of cocaine CPP, respectively. To investigate consolidation, rats received intra-DHC muscimol either immediately or 6 hr after conditioning sessions. Inhibition of DHC, but not the overlying cortex, disrupted acquisition and expression of cocaine CPP. It is interesting to note that there was no effect of post-conditioning DHC inhibition. The findings suggest that the DHC is important for both acquisition and recall, but not consolidation, of context-cocaine associations. PMID- 16719705 TI - Performance on spatial working memory tasks after dorsal or ventral hippocampal lesions and adjacent damage to the subiculum. AB - Rats with excitotoxic lesions of the dorsal or ventral hippocampus and control rats were trained on 2 spatial working memory tasks: the standard version of the radial maze with 8 baited arms and the non-matching-to-place procedure in the T maze. Dorsal lesions produced deficits in both tasks, whereas ventral lesions did not affect learning in either of them. A volumetric analysis of subicular damage showed that dorsal hippocampal lesions caused a deficit in the non-matching-to place only when accompanied by damage to the dorsal subiculum; on the other hand, lesions to the dorsal hippocampus impaired performance in the radial-arm maze regardless of the extent of subicular damage. PMID- 16719706 TI - Subdivisions of inferior temporal cortex in squirrel monkeys make dissociable contributions to visual learning and memory. AB - Inferior temporal cortex of squirrel monkeys consists of caudal (ITC), intermediate (ITI), and rostral (ITR) subdivisions, possibly homologous to TEO, posterior TE, and anterior TE of macaque monkeys. The present study compared visual learning in squirrel monkeys with ablations of ITC; ITI and ITR (group ITRd); or ITI, ITR, and more ventral cortex, including perirhinal cortex (group ITR+), with visual learning in unoperated controls. The ITC monkeys had significant impairments on pattern discriminations and milder deficits on delayed non-matching to sample (DNMS) of objects. The ITRd monkeys had deficits on some pattern discriminations but not on DNMS. The ITRd monkeys were significantly impaired on DNMS and some pattern discriminations. These results are similar to those found in macaques and support the proposed homologies. PMID- 16719707 TI - The ventral premotor cortex (vPM) and resistance to interference. AB - The authors tested a patient suffering from a circumscribed lesion of the right frontal operculum (FO) in 3 experiments of visual attention involving spatial orienting, maintenance of task-relevant priorities, and control of interference from new and old task-irrelevant items. The authors found that spatial orienting and active maintenance of priorities were intact, but there were difficulties in controlling interference from new and old irrelevant items. These results suggest that the FO is necessary for the direct control of interference, but its lesion alone is not enough to disturb spatial orienting processes or active maintenance of task priorities. The authors discuss the results in light of a hybrid cognitive model of attention. PMID- 16719708 TI - Differences in the sensations of cold on the anterior torso of control and spinal cord-transected individuals. AB - Application of cold stimuli to the sentient portion of the anterior torso of 12 spinal-cord-transected individuals (patients) and to comparable sites of 11 control participants showed that thresholds are lower for women than for men and that the difference between the sexes is maintained following spinal-cord transection. Patients of both sexes were more sensitive to cold stimuli than were controls. Estimates of stimulus intensity showed that participants reliably distinguished the 3 cool stimuli but that control women offered significantly larger estimates than control men. Spinal-cord transection produced an increase in the intensity of the sensations in women and a reduction in men. The changes in sensory perception that follow spinal-cord injury extend throughout the somatosensory system and involve all modalities. These changes cannot be explained as a simple release from inhibition. PMID- 16719709 TI - Reduced variability of postural strategy prevents normalization of motor changes induced by back pain: a risk factor for chronic trouble? AB - Variability is fundamental to biological systems and is important in posturomotor learning and control. Pain induces a protective postural strategy, although variability is normally preserved. If variability is lost, does the normal postural strategy return when pain stops? Sixteen subjects performed arm movements during control trials, when the movement evoked back pain and then when it did not. Variability in the postural strategy of the abdominal muscles and pain-related cognitions were evaluated. Only those subjects for whom pain induced a reduction in variability of the postural strategy failed to return to a normal strategy when pain stopped. They were also characterized by their pain-related cognitions. Ongoing perception of threat to the back may exert tighter evaluative control over variability of the postural strategy. PMID- 16719710 TI - Sustained attention in the mouse: a study of the relationship with the cerebellum. AB - To explore the role of the cerebellum in sustained attention, the authors tested lurcher, wildtype, and lurcher chimeric mice with zero, normal, and variable numbers of Purkinje cells, respectively, in a previously validated task of sustained attention. Results indicate that lurcher mice had a deficit in performance likely related to their motor disability, whereas lurcher chimeras performed similarly to wildtype controls. Presentation of auditory or visual distracters caused deficits in the performance of all mice that were specific to either signal (auditory) or non-signal (visual) events. The authors' results do not support a role of the cerebellum in sustained attention, instead indicating that behavioral changes are an indirect result of motor deficits. PMID- 16719711 TI - Impaired trace fear conditioning following neonatal ethanol: reversal by choline. AB - Neonatal ethanol exposure in animals results in performance deficits on tests of hippocampus-dependent spatial memory, and recent studies have shown that extra dietary choline can ameliorate some of these impairments. In this experiment, rats were administered 5.25 g/kg ig ethanol per day or sham intubations on Postnatal Days (PD) 4-9 and choline (0.1 ml of an 18.8 mg/ml solution) or saline subcutaneously on PD 4-20. On PD 30, rats were given delay or trace fear conditioning trials and were tested for conditioned stimulus-elicited freezing 24 hr later. Neonatal ethanol produced a profound impairment in trace conditioning that was reversed by choline. Groups did not differ in delay conditioned responding, indicating that neonatal ethanol produces a relatively selective cognitive deficit that can be alleviated with supplemental choline. PMID- 16719712 TI - Dopamine receptor blockade and extinction differentially affect behavioral variability. AB - Dopamine (DA) neurons respond to unexpected food delivery and are inhibited during the omission of expected reward. DA receptor blockade mimics some, but not all, aspects of non-reward (extinction) conditions. It was therefore of interest to ask whether DA receptor blockade produces extinction-like increases in behavioral variability in addition to its well-known operant response-suppressing effects. In the current experiment, rats were trained drug-free on an operant task in which they pressed on a keyboard. Two of the keys led to food on a continuous reinforcement schedule. Both response rates and behavioral variability were measured. Test day administration of D(1) and D(2) antagonists SCH23390 and raclopride, like extinction, suppressed responding but, unlike extinction, did not lead to an increase in variability. PMID- 16719713 TI - The consequences of atheoretical, task-driven experimentation: theoretical comment on Paban, Chambon, Jaffard, and Alescio-Lautier (2005). AB - V. Paban, C. Chambon, M. Jaffard, and B. Alescio-Lautier (2005) concluded that their evidence "demonstrates the independence of aging and cholinergic lesion effects" and stated that "this was quite unexpected" (p. 943). This commentary suggests that the authors' negative findings were predictable, resulting from a focus on behavioral tests and functions unrelated to present evidence and theory concerning the cognitive functions of forebrain cholinergic systems. The article by V. Paban et al. serves as a useful illustration for the importance of theory- and hypothesis-driven assessment of behavioral functions, and for the unproductive consequences of experiments treating behavioral tests as assays used merely for the generation of behavioral correlates. PMID- 16719715 TI - Environmental stress and lesion-bypass DNA polymerases. AB - In nature, microbes live under a variety of harsh conditions, such as excess DNA damage, starvation, pH shift, or high temperatures. Microbial cells respond to such stressful conditions mostly by switching global patterns of gene expression to relieve the environmental stress. The SOS response, which is induced by DNA damage, is one such global network of gene expression that plays a crucial role in balancing the genomic stability and flexibility that are necessary to adapt to harsh environments. Here, I review the roles of SOS-inducible and noninducible lesion-bypass DNA polymerases in mutagenesis induced by environmental stress, and discuss how these polymerases are coordinated for the replication of damaged chromosomes. Possible contributions of lesion-bypass DNA polymerase in hyperthermophilic archaea, e.g., Sulfolobus solfataricus, to genome maintenance are also discussed. PMID- 16719716 TI - Environmental biology of the marine Roseobacter lineage. AB - The Roseobacter lineage is a phylogenetically coherent, physiologically heterogeneous group of alpha-Proteobacteria comprising up to 25% of marine microbial communities, especially in coastal and polar oceans, and it is the only lineage in which cultivated bacteria are closely related to environmental clones. Currently 41 subclusters are described, covering all major marine ecological niches (seawater, algal blooms, microbial mats, sediments, sea ice, marine invertebrates). Members of the Roseobacter lineage play an important role for the global carbon and sulfur cycle and the climate, since they have the trait of aerobic anoxygenic photosynthesis, oxidize the greenhouse gas carbon monoxide, and produce the climate-relevant gas dimethylsulfide through the degradation of algal osmolytes. Production of bioactive metabolites and quorum-sensing-regulated control of gene expression mediate their success in complex communities. Studies of representative isolates in culture, whole-genome sequencing, e.g., of Silicibacter pomeroyi, and the analysis of marine metagenome libraries have started to reveal the environmental biology of this important marine group. PMID- 16719717 TI - Structure and function of RNA replication. AB - Contrary to their host cells, many viruses contain RNA as genetic material and hence encode an RNA-dependent RNA polymerase to replicate their genomes. This review discusses the present status of our knowledge on the structure of these enzymes and the mechanisms of RNA replication. The simplest viruses encode only the catalytic subunit of the replication complex, but other viruses also contribute a variable number of ancillary factors. These and other factors provided by the host cell play roles in the specificity and affinity of template recognition and the assembly of the replication complex. Usually, these host factors are involved in protein synthesis or RNA modification in the host cell, but they play roles in remodeling RNA-RNA, RNA-protein, and protein-protein interactions during virus RNA replication. Furthermore, viruses take advantage of and modify previous cell structural elements, frequently membrane vesicles, for the formation of RNA replication complexes. PMID- 16719718 TI - Transcriptional regulatory elements in the human genome. AB - The faithful execution of biological processes requires a precise and carefully orchestrated set of steps that depend on the proper spatial and temporal expression of genes. Here we review the various classes of transcriptional regulatory elements (core promoters, proximal promoters, distal enhancers, silencers, insulators/boundary elements, and locus control regions) and the molecular machinery (general transcription factors, activators, and coactivators) that interacts with the regulatory elements to mediate precisely controlled patterns of gene expression. The biological importance of transcriptional regulation is highlighted by examples of how alterations in these transcriptional components can lead to disease. Finally, we discuss the methods currently used to identify transcriptional regulatory elements, and the ability of these methods to be scaled up for the purpose of annotating the entire human genome. PMID- 16719719 TI - Evolution of plant pathogenicity in Streptomyces. AB - Among the multitude of soil-inhabiting, saprophytic Streptomyces species are a growing number of plant pathogens that cause economically important diseases, including potato scab. Streptomyces scabies is the dominant pathogenic species worldwide, but is only one of many that cause very similar disease symptoms on plants. Molecular genetic analysis is beginning to identify the mechanisms used by plant pathogenic species to manipulate their hosts. The nitrated dipeptide phytotoxin, thaxtomin, inhibits cellulose biosynthesis in expanding plant tissues, stimulates Ca2+ spiking, and causes cell death. A secreted necrogenic protein, Nec1, contributes to virulence on diverse plant species. The thaxtomin biosynthetic genes and nec1 lie on a large mobilizable PAI, along with other putative virulence genes including a cytokinin biosynthetic pathway and a saponinase homolog. The PAI is mobilized during conjugation and site-specifically inserts in the linear chromosome of recipient species, accounting for the emergence of new pathogens in agricultural systems. The recently available genome sequence of S. scabies will accelerate research on host-pathogen interactions. PMID- 16719720 TI - Phenazine compounds in fluorescent Pseudomonas spp. biosynthesis and regulation. AB - The phenazines include upward of 50 pigmented, heterocyclic nitrogen-containing secondary metabolites synthesized by some strains of fluorescent Pseudomonas spp. and a few other bacterial genera. The antibiotic properties of these compounds have been known for over 150 years, but advances within the past two decades have provided significant new insights into the genetics, biochemistry, and regulation of phenazine synthesis, as well as the mode of action and functional roles of these compounds in the environment. This new knowledge reveals conservation of biosynthetic enzymes across genera but raises questions about conserved biosynthetic mechanisms, and sets the stage for improving the performance of phenazine producers used as biological control agents for soilborne plant pathogens. PMID- 16719722 TI - Targeting norm-related beliefs about marijuana use in an adolescent population. AB - The integrative model of behavior prediction and priming theory were used to evaluate the effects of anti-marijuana advertisements in an experimental context. In 1 original study and 2 replications, 435 adolescents were randomly assigned to condition, and those in the experimental condition viewed 3 ads that challenged undesirable normative beliefs about marijuana use. The results showed that ad exposure had small but positive (anti-drug) effects on adolescents' considerations of the outcomes of using marijuana and their perceptions of the social normative climate surrounding marijuana use. Priming effects also were observed but generally ran counter to predictions. Implications of the findings for anti-drug campaigns are discussed. Potential explanations for the unexpected priming effects also are offered. PMID- 16719723 TI - Transforming emergency medicine through narrative: qualitative action research at a community hospital. AB - Emergency departments struggle daily to save lives in an environment characterized by staff shortages, limited resources, and an expanding patient population. This qualitative action research study focused on the nature of communication in an urban emergency room (ER) and the organizing practices employed by staff to cope with these environmental changes, highlighting disconnects between current practices and traditional models of emergent care. A narrative description of the ER culture served as an impetus for practical improvements at this site, providing staff with both a unique perspective and a useful tool for improving their emergency care practices. PMID- 16719724 TI - Differential effects of exposure to social norms campaigns: a cause for concern. AB - College students' processing of alcohol social norms messages, related effects on normative judgments, attitudes toward their own behaviors, and perception of undergraduate attitudes were examined using expectancy violation theories and social norms marketing. Data were collected from 2 universities (N = 393). Following message exposure, the majority moved their normative judgments toward the statistic provided in the message. Slight attitude change occurred but not always in the desired direction. Those most likely to develop unhealthier attitudes drank more than those who developed healthier attitudes, consistent with psychological reactance to the messages. Therefore, the effects of social norms campaigns on those at greatest risk for primary and secondary alcohol effects due to their increased alcohol consumption could lead to increased risk for those participants, indicating that the widespread use of social norms campaigns needs to be scrutinized. PMID- 16719725 TI - Parent-therapist communication and satisfaction with the services of a child development center: a comparison between Israeli parents-Jews and Bedouins. AB - This study evaluates the level of general satisfaction with the services of a child development center in 2 groups of Israeli parents, Jews and Bedouins. It also assesses which 3 dimensions of therapist-parent communication significantly explain general satisfaction with the center's services. A total of 193 parents of children ranging from 6 months to 6 years of age participated in the study (84% response rate). Parents' attitudes regarding their communication with the center's therapists were evaluated by 15 items that were clustered into 3 dimensions of communication: caring, collaboration, and interest. General satisfaction with the center's services was high in both groups. Relatively high average scores were also given to the studied dimensions of communication. Jews were more satisfied with the center's services than were Bedouins and ranked caring and collaboration significantly higher than did Bedouins. Caring and collaboration best explained the variability in general satisfaction in a multivariate analysis. Being a Jew or a Bedouin parent was not found to be a significant explanatory variable of general satisfaction with the center's services. Therapists' expressions of care and collaboration with parents seem to be key factors in explaining parents' general satisfaction with rehabilitation services in the 2 different cultural groups. These factors should therefore be nurtured in order to ensure parents' satisfaction and participation in the ongoing and demanding process of child rehabilitation. The question of whether this finding is applicable to other populations has to be further studied. PMID- 16719726 TI - Increasing selective exposure to health messages by targeting person versus behavior schemas. AB - Schema correspondence theory (Brannon and Brock, 1994) was applied to the topic of selective exposure to health information. The following question was asked: When do people prefer to expose themselves to health-relevant information tailored to match their own needs and values (i.e., recipient self-schema matching) versus the values and goals that the healthy behavior brings to mind (i.e., behavior schema matching)? In general, recipient self-schema matched messages tended to be preferred over behavior schema matched messages. However, this tendency was attenuated to the extent that the behavior had a very well defined (prototypical) schema. PMID- 16719727 TI - Identifying principal risk factors for the initiation of adolescent smoking behaviors: the significance of psychological reactance. AB - An in-school youth survey for a major state anti-tobacco media campaign was conducted with 1,831 students (Grades 6-12) from 70 randomly selected classrooms throughout the state. Tobacco users accounted for nearly 25% of the sample. Pretest questionnaires assessed demographic variables, tobacco use, and various other risk factors. Several predictors of adolescents' susceptibility to tobacco use, including prior experimentation with tobacco, school performance, parental smoking status, parents' level of education, parental communication, parental relationship satisfaction, best friend's smoking status, prevalence of smokers in social environment, self-perceived potential to smoke related to peer pressure, and psychological reactance, were examined using discriminant analysis and logistic regression to identify the factors most useful in classifying adolescents as either high-risk or low-risk for smoking uptake. Results corroborate findings in the prevention literature indicating that age, prior experimentation, and having friends who smoke are among the principal predictors of smoking risk. New evidence is presented indicating that psychological reactance also should be considered as an important predictor of adolescent smoking initiation. The utility of producing antismoking messages informed by an awareness of the key risk factors-particularly psychological reactance-is discussed both in terms of the targeting and design of anti-tobacco campaigns. PMID- 16719728 TI - Effects of exposure of youths at risk for smoking to television advertising for nicotine replacement therapy and Zyban: an experimental study. AB - Television advertising for nicotine replacement therapy (NRT) and Zyban exposes the entire population, including adolescents, to persuasive messages about these smoking-cessation products. There is a risk that adolescents exposed to the advertising might underestimate addictiveness or perceive an unintended message that it is easy to quit smoking. This is of concern because optimism about quitting is a major predictor of trial and progression to heavier smoking among youths. We randomly allocated 492 youths age 12 to 14 years to one of three viewing conditions in which they viewed either (a) 4 NRT ads, (b) 4 Zyban ads, or (c) 4 ads promoting nonpharmacologic cessation services, such as telephone quitlines. After viewing each ad twice, participants completed a 1-page rating form. After all ads had been viewed, youths completed a questionnaire that measured intentions to smoke in the future, perceived addictiveness of smoking, perceived risks and benefits of smoking, and perceived need for pharmaceutical products and services. There were no differences in the composition of groups by age, gender, or smoking uptake. Youths were more likely to agree that the NRT and Zyban ads, compared with the quitline ads, made it seem easy to quit smoking (p < .001). However, there were no systematic differences between groups in perceived addictiveness of smoking, intentions to smoke, or other outcomes. This study suggests that although ads for NRT and Zyban may create "face value" impressions that it is easier to quit, at least in an experimental context in which exposure to ads for telephone quitlines is equal, such appraisals do not undermine more enduring perceptions about smoking. Field research taking into account the relatively high volume of pharmaceutical cessation product advertising is needed. PMID- 16719729 TI - Compliance and intimacy: young adults' attempts to motivate health-promoting behaviors by romantic partners. AB - Benefits of relational intimacy on health may derive from its influence on the success of verbal attempts to motivate romantic partners' compliance with suggested protective behaviors. The intimacy levels of 209 participants' romantic relationships were assessed, as were motivations to change 4 health behaviors in response to 8 appeal strategies. Intimacy had significant impacts on the estimated effectiveness of several appeals. Also, as intimacy increased, caring replaced the provision of health information as the most efficacious strategy for motivating dietary modification. Overall, appeals seeking safer sex and those employing liking, caring, and threat strategies were rated as generally most effective. PMID- 16719731 TI - Bibliography. PMID- 16719730 TI - The case of the missing person: Alzheimer's disease in mass print magazines 1991 2001. AB - Alzheimer's disease is growing in incidence and prevalence in the developed world. Rates have been increasing as populations have been aging. There are still many unknowns regarding prevention, causes, and treatments. The purpose of this article is to analyze the portrayal of Alzheimer's in the highest-circulation mass print English-language magazines published in the United States and Canada over a period of a decade, specifically those for 1991, 1996, and 2001. This research investigates the portrayal of persons with Alzheimer's, the disease itself, caregivers and experts, and the dominant frames or discourses within which Alzheimer's is described. Twenty-five articles from the highest-circulation mass print magazines available in Canada were studied through qualitative and inductive research of both manifest and latent content. One of the most notable findings is the absence of the person with the disease as a person with a voice, with needs and desires. When the disease itself is described it is characterized as fearsome, relentless, and aggressive. Both the unquestioned duty of the individual caregiver and his or her (usually the caregiver is a woman) suffering are emphasized. The disease, its diagnosis, and potential treatments are framed almost exclusively within a medical rather than a political-economy or lifestyle frame. Minimal attention is paid to prevention, early stages of the disease, social support, options for care, or other alternative understandings of issues related to Alzheimer's. The theoretical and practical significance of these findings is considered. PMID- 16719733 TI - The safety and quality of health care: from Council to Commission. PMID- 16719734 TI - Shared meanings: preferred terms and definitions for safety and quality concepts. PMID- 16719735 TI - Sea change: public reporting and the safety and quality of the Australian health care system. AB - The pursuit of demonstrable safety and quality in health care is an evolving process; there has been notable progress in measuring safety and quality in Australia. The first attempts to measure outcomes were in the field of anaesthesia, while national perinatal mortality reports have provided clinically useful information for many years. Nationwide reporting by the Quality in Australian Health Care Study (QAHCS) in 2005 triggered a more systemic approach to safety and quality. Systemic reporting has begun to emerge in anaesthesia and surgery, for implantable devices, perinatal services and sentinel events; in some jurisdictions, statewide incident data are now reported annually. While debate continues about the issue of individual clinician performance, the real issue is the effectiveness of any reporting system to bring about change in both safety and quality. PMID- 16719736 TI - Health care safety and quality: where have we been and where are we going? AB - Health care will always be associated with risk, but the Australian Council for Safety and Quality in Health Care has achieved much in bringing health care safety and quality into public consciousness and beginning systemic change for improvement. Work is underway to develop safety and quality standards, and infrastructure and systems for measurement and evaluation; to increase workforce understanding of how to improve health care delivery; to increase consumer engagement in health care management; and to develop policy and understanding of the barriers to progress. With this foundation of reform, the future of the new Australian Commission on Safety and Quality in Health Care is promising, but it is up to us as health professionals and managers, with the help of the community, to improve the safety and quality of health care. PMID- 16719737 TI - Measurement of the safety and quality of health care. AB - Measurement of safety and quality is fundamental to health care delivery. A variety of measures are needed to fully understand the system; quantitative and qualitative measures are both useful in different ways. Measures need to be valid, reliable, accurate, timely, collectable, meaningful, relevant and important to those who will use them. Clinicians value appropriate measures and respond to them. PMID- 16719738 TI - Designing safer health care through responsive regulation. AB - Self-regulation by the health professions, while improving, is no longer enough; external drivers for safer health care include governments, funders and consumers. Enforced self-regulation is often more promising than a "command and control" strategy. Research evidence on the responsive regulatory pyramid and its options offers lessons for health care policy makers and managers. Start at the base of the regulatory pyramid - try persuasion first; move up the pyramid to secure compliance, and then be willing to move back down. Use existing capacities and structures, and if possible avoid new bureaucracies of control. PMID- 16719739 TI - Improving safety and quality: how can education help? AB - National efforts to improve the quality and safety of health care present challenges for medical education and training. Today's doctors need to be skilled communicators who know how to identify, prevent and manage adverse events and near misses, how to use evidence and information, how to work safely in a team, how to practise ethically, and how to be workplace teachers and learners. These competencies (knowledge, skills and attitudes) are set out in the National Patient Safety Education Framework (NPSF) of the Australian Council for Safety and Quality in Health Care. The NPSF is designed to help medical schools, vocational colleges, health organisations and private practitioners develop curricula to enable health professionals to work safely. The NPSF describes what doctors (depending on their level of knowledge and experience) can do to demonstrate competencies in a range of quality and safety activities. Medical schools, vocational colleges, health organisations and private practitioners need to work collaboratively with one another and with other health professionals to ensure that patient safety and quality curricula are implemented and evaluated, and that valid and reliable assessments of learning outcomes are developed. Interdisciplinary and vertically integrated education and training are needed, incorporating innovative methods, to create a safer health care system. PMID- 16719740 TI - The development of the National Reporting and Learning System in England and Wales, 2001-2005. AB - In 2001, the National Patient Safety Agency (NPSA) was created as part of a wider reform process to improve quality of care for patients in the National Health Services of England and Wales. The NPSA was charged with developing and implementing a national system for collecting and learning from reported patient safety incidents. Between 2003 and 2005, 303 447 incidents were reported from a wide range of health care settings. As a result, a range of interventions have been developed to improve safety. A number of lessons have been distilled from the experience of England and Wales, including that: clinical risk management system characteristics should be aligned with those of the national reporting system; and safety culture and information dissemination must be addressed at the same time as any new reporting system is implemented. These lessons should be of use to other countries implementing similar patient safety strategies. PMID- 16719741 TI - The WHO World Alliance for Patient Safety: towards the years of living less dangerously. AB - Improving the safety of patient care is now a global issue. In 2004, the World Health Assembly supported the creation of the World Alliance for Patient Safety to coordinate, spread and accelerate improvements in patient safety internationally. Australia has been at the forefront of international work on patient safety and is working closely with the World Alliance. The World Alliance is taking forward work in six main action areas: Patients for Patient Safety; Reporting and Learning; Taxonomy; Solutions; Research; and Global Patient Safety Challenge. Despite differences in context, there are many common challenges for countries seeking to establish patient safety programs and initiatives. Looking forward, long-term action on patient safety must be built and sustained with the commitment of policy makers and the active engagement of health care professionals. PMID- 16719742 TI - Indigenous health: burden or opportunity? PMID- 16719743 TI - Strengthening cardiac rehabilitation and secondary prevention for Aboriginal and Torres Strait Islander peoples. PMID- 16719744 TI - Better late than never: a national approach to trachoma control. PMID- 16719745 TI - Causes of inequality in life expectancy between Indigenous and non-Indigenous people in the Northern Territory, 1981-2000: a decomposition analysis. AB - OBJECTIVE: To identify the causes of the gap in life expectancy between Indigenous and non-Indigenous populations of the Northern Territory and how the causes have evolved over time. DESIGN AND SETTING: Analysis of NT death data over four 5-year periods, 1 January 1981 to 31 December 2000 inclusive. A decomposition method using discrete approximations (Vaupel and Romo) was applied to abridged life tables for the Indigenous and non-Indigenous populations of the NT. MAIN OUTCOME MEASURES: Contribution of causes of death, grouped according to global burden of disease groups and categories, to the life expectancy gap. RESULTS: The gap between the life expectancy of Indigenous and non-Indigenous people in the NT did not appear to narrow over time, but there was a marked shift in the causes of the gap. In terms of disease groups, the contribution of communicable diseases, maternal, perinatal and nutritional conditions halved during the 20 years to 2000. Meanwhile, the contribution of non-communicable diseases and conditions increased markedly. The contribution of injuries remained static. In terms of disease categories, the contribution of infectious diseases, respiratory infections and respiratory diseases declined considerably; however, these gains were offset by significantly larger increases in the contribution of cardiovascular diseases and diabetes for Indigenous women and cardiovascular diseases, cancers and digestive diseases for Indigenous men. CONCLUSIONS: The main contributors to the gap in life expectancy between the Indigenous and non Indigenous populations were non-communicable diseases and conditions, which are more prevalent in ageing populations. With the life expectancy of Indigenous people in the NT expected to improve, it is important that public health initiatives remain focused on preventing and managing chronic diseases. PMID- 16719746 TI - The outcome of critically ill Indigenous patients. AB - OBJECTIVE: To investigate the short-term outcome of critically ill Indigenous patients. DESIGN AND PARTICIPANTS: Retrospective cohort study using de-identified audit data from a tertiary intensive care unit (ICU) in Western Australia for the 11-year period 1 January 1993 to 31 December 2003. MAIN OUTCOME MEASURES: Hospital mortality (crude, and adjusted for severity of illness). RESULTS: Of 16 757 ICU patients, 1076 (6.4%) were identified as Indigenous. The Indigenous patients were younger and more commonly had chronic liver and renal diseases. Indigenous people represented 3.2% of the population of Western Australia in 2001, but represented 3.1% and 9.5% of all elective and emergency ICU admissions, respectively. Diagnoses of sepsis, pneumonia, trauma, and cardiopulmonary arrest were common among critically ill Indigenous patients. Following emergency admission, the crude hospital mortality for Indigenous patients was higher (22.7% v 19.2%; crude odds ratio, 1.24; 95% CI, 1.04-1.47) than for non-Indigenous patients. The crude hospital mortality of critically ill Indigenous patients was lower than that predicted by the APACHE II prognostic model and was similar to that of non-Indigenous patients after adjusting for severity of illness and chronic health status. CONCLUSIONS: The pattern of critical illness affecting Indigenous Australians in Western Australia was different from that affecting non Indigenous patients. The crude hospital mortality was high, but similar to that of non-Indigenous Australians after adjusting for severity of illness and chronic health status. PMID- 16719747 TI - Getting the most from Indigenous health research. PMID- 16719748 TI - Indigenous health research: a critical review of outputs over time. AB - OBJECTIVE: To determine the number and nature of publications on Indigenous health in Australia, Canada, New Zealand and the United States) in 1987-1988, 1997-1998 and 2001-2003. DATA SOURCES: MEDLINE and PsychLit databases were searched using the following terms: Aborigines or Aboriginal; Torres Strait Islander; Maori; American Indian; North American Indian, or Indian, North American; Alaska/an Native; Native Hawaiian; Native American; American Samoan; Eskimos or Inuit; Eskimos or Aleut; Metis; Indigenous. STUDY SELECTION: Publications were included if they were concerned with the health of Indigenous people of the relevant countries. 1763 Indigenous health publications were selected. DATA EXTRACTION: Publications were classified as either: original research; reviews; program descriptions; discussion papers or commentaries; or case reports. Research publications were further classified as either measurement, descriptive, or intervention. Intervention studies were then classified as either experimental or non-experimental. DATA SYNTHESIS: The total number of publications was highest in 1997-1998 for most countries. The most common type of publication across all time periods for all countries was research publications. In Australia only, the number of research publications was slightly higher in 2001-2003 compared with other time periods. For each country and at each time, research was predominantly descriptive (75%-92%), with very little measurement (0-11%) and intervention research (0-18%). Overall, of the 1131 research publications, 983 were descriptive, 72 measurement and 76 intervention research. CONCLUSIONS: The dominance of descriptive research in Indigenous health is not ideal, and our findings should be carefully considered by research organisations and researchers when developing research policies. PMID- 16719749 TI - Acute rheumatic fever and rheumatic heart disease: an insight into Aboriginal health disadvantage and remote Australia. PMID- 16719750 TI - B-cell antigen D8/17 is a marker of rheumatic fever susceptibility in Aboriginal Australians and can be tested in remote settings. AB - OBJECTIVE: To test the B-cell antigen D8/17 as a marker of past rheumatic fever (RF) in a predominantly Aboriginal Australian population, and to evaluate technical modifications to allow its use in remote settings. DESIGN AND SETTING: Cross-sectional survey in a remote Aboriginal community, a regional tertiary referral hospital and a tertiary paediatric centre in Melbourne. PARTICIPANTS: 106 people, including three with acute RF, 38 with a history of past RF, 20 relatives of these people, and 45 healthy controls. MAIN OUTCOME MEASURE: D8/17 expression in B cells. RESULTS: Blood was collected from each participant and the expression of D8/17 and CD19 in each sample was analysed by flow cytometry. The mean proportion of D8/17-positive B cells was 39.3% (SD, 11.8) in patients with previous RF, 22.5% (SD, 5.2) in first-degree relatives, 11.6% (SD, 7.2) in controls, and 83.7% (SD, 10.1) in patients with acute RF (analysis of variance test between means, P = 0.001). A cut-off of 22.1% of D8/17-positive B cells to indicate past RF yielded the highest percentage of correct results (95.4%). Delayed staining of whole blood (mean, 0.55 days; SD, 0.2) gave equivalent results to immediate staining, but the D8/17 assay on peripheral blood mononuclear cells was unreliable. CONCLUSIONS: The B-cell antigen D8/17 accurately identifies Australians with a past history of RF, and the assay is feasible in remote settings with access to facilities capable of performing D8/17 staining within half a day of sample collection. PMID- 16719751 TI - Practical challenges of conducting research into rheumatic fever in remote Aboriginal communities. AB - Before embarking on an epidemiological study of acute rheumatic fever in remote Aboriginal communities, researchers engaged in the processes of community consultation, consent and household enrollment. Community expectations and time constraints are not necessarily those of the funding bodies, and a considerable investment of time and local engagement was required before the project proceeded with local support. The remoteness of the communities, harsh climate and limited infrastructure made working conditions difficult. Nevertheless, the study was completed and the results are being returned to the local councils and households. The research team continues to maintain its relationship with each study community. PMID- 16719753 TI - Mental health, grief and family ties. The Dr Ross Ingram Memorial Essay Competition 2006. PMID- 16719752 TI - Challenging perceptions of non-compliance with rheumatic fever prophylaxis in a remote Aboriginal community. AB - AIM: To identify factors that affect rheumatic fever prophylaxis for remote living Aboriginal patients, and to determine the proportion who received adequate prophylaxis. DESIGN AND SETTING: Interview (with analysis based on principles of grounded theory) of patients with a history of rheumatic fever or rheumatic heart disease and their relatives, and health service providers in a remote Aboriginal community; audit of benzathine penicillin coverage of patients with rheumatic heart disease. PARTICIPANTS: 15 patients with rheumatic heart disease or a history of rheumatic fever, 18 relatives and 18 health care workers. RESULTS: Patients felt that the role of the clinic was not only to care for them physically, but that staff should also show nurturing holistic care to generate trust and treatment compliance. Differing expectations between patients and health care providers relating to the responsibility for care of patients absent from the community was a significant factor in patients missing injections. Neither a biomedical understanding of the disease nor a sense of taking responsibility for one's own health were clearly related to treatment uptake. Patients did not generally refuse injections, and 59% received adequate prophylaxis (> 75% of prescribed injections). CONCLUSION: In this Aboriginal community, concepts of being cared for and nurtured, and belonging to a health service were important determinants of compliance. PMID- 16719754 TI - Unknown family at the taxi stand. The Dr Ross Ingram Memorial Essay Competition 2006. PMID- 16719755 TI - Making a difference: the early impact of an Aboriginal health undergraduate medical curriculum. AB - OBJECTIVE: To describe the implementation of an integrated Aboriginal health curriculum into the medical course at the University of Western Australia (UWA) and the early effect on students' perceptions of their knowledge and ability in the area of Aboriginal health. DESIGN, SETTING AND PARTICIPANTS: Final-year medical students at UWA in 2003 (first cohort) and 2004 (second cohort) were surveyed by questionnaire (with answers on a five-point Likert scale) to assess their attitudes to various aspects of Aboriginal health. A subset of students provided open-ended comments on key priorities in Aboriginal health, cultural security and suggestions for Aboriginal health policy. INTERVENTIONS: Integrated learning experiences were implemented within each year of the medical course, based on specific learning outcomes in Aboriginal health. MAIN OUTCOME MEASURES: Changes in students' self-perceptions of their preparedness for and future commitment to working for change in Aboriginal health. RESULTS: Response rates were 76% and 85% in the 2003 and 2004 cohorts, respectively. Compared with first cohort students, second-cohort students were more likely to agree with items relating to their preparedness and ability to work with and care for Aboriginal and Torres Strait Islander people (P < 0.05); second-cohort students also reported greater preparedness to advocate and improve the health of Aboriginal people (P < 0.05); 65% of respondents in the second cohort (versus 34% in the first) agreed they had a social responsibility to work for change in Aboriginal health (P < 0.05). CONCLUSION: With a relatively small amount of targeted and structured teaching and learning in Aboriginal health, significant shifts in students' self-perceived levels of knowledge, skills and attitudes are possible. PMID- 16719756 TI - Perspectives on Aboriginal community controlled health services. PMID- 16719757 TI - The general practitioner. PMID- 16719758 TI - The Aboriginal health worker. PMID- 16719759 TI - The manager. PMID- 16719760 TI - The importance of Hand Talk in communication rehabilitation among Aboriginal Australians in the Northern Territory. PMID- 16719761 TI - Emphysematous pyelonephritis. PMID- 16719762 TI - Aboriginal incarceration: health and social impacts. AB - Each year up to a quarter of all young Aboriginal men have direct involvement with correctional services, and Aboriginal prisoners currently represent 22% of the total Australian prisoner population. The high rates of repeated short-term incarceration experienced by Aboriginal people in Australia have a multitude of negative health effects for Aboriginal communities and the wider society, while achieving little in terms of increased community safety. Well identified health and social priorities for Aboriginal people affected by incarceration include housing and tenancy support; mental health and wellbeing, including family violence, grief and loss support; substance misuse support; general health services, including hepatitis C management; and social inclusion, including the need for family and community integration, skills development and employment. The post-release period is a crucial time for the provision of integrated health and social services to address these priorities and to break the cycle of incarceration. To achieve significant health gains for Aboriginal people, there is a need to develop a broader collaborative approach to primary health care, incorporating social health and justice perspectives as fundamental components of health care planning. Health and human services have a critical role to play in developing community-based solutions to reduce excessive incarceration rates for Aboriginal people. PMID- 16719764 TI - Airway smooth muscle phenotype and function: interactions with current asthma therapies. AB - Asthma incidence has climbed markedly in the past two decades despite an increased use of medications that suppress airway inflammation and repress contraction of smooth muscle that encircles the airways. Asthmatics exhibit episodes of airway inflammation that potentiates reversible airway smooth muscle spasm. A hallmark diagnostic symptom of asthma is airway hyperresponsiveness to inhaled non-allergic stimuli, such as methacholine, that directly induce airway smooth muscle contraction. Inhaled gluccocorticoids are used for first-line prevention of airway inflammation, and are frequently combined with inhaled beta2 adrenoceptor agonists that can effectively relax airway smooth muscle and restore airway conductance. Leukotriene receptor antagonists and anti-cholinergics can also be used in many patients to ensure optimal control of symptoms. With increasing disease duration irreversible airway restriction develops from inflammation-driven fibro-proliferative airway remodeling that includes increased deposition of extracellular matrix, the accumulation of airway smooth muscle, and increased numbers of myofibroblasts. Mature airway smooth muscle cells are phenotypically plastic, enabling them to subserve contractile, proliferative, migratory and secretory functional responses that contribute to airway remodeling and persistent hyperresponsiveness. This review assesses current understanding of acute and chronic effects of common anti-asthma medications on the diverse phenotype and functional characteristics of airway smooth muscle cells. Furthermore, we describe the significance of these effects in the treatment of asthma symptoms and pathogenesis. PMID- 16719765 TI - Interactions between airway epithelial cells and dendritic cells: implications for the regulation of airway inflammation. AB - Airway epithelial cells (AEC) and dendritic cells (DC) are situated in close proximity within the airway epithelium, and are the first cells to encounter inhaled pathogens, allergens and environmental pollutants. AEC and DC interact through the release of cytokines and other soluble mediators and through direct cell-cell contact, and these interactions are likely to play an important role in maintaining immune homeostasis. Increasing evidence indicates that both AEC and DC from asthmatic individuals exhibit distinct functional properties, compared with AEC and DC from healthy individuals. Both animal models, and novel co culture models for directly studying human AEC/DC interactions are providing new insight into the cross-talk between these two important cells types, as a foundation for the development of new drug targets for use in asthma, cystic fibrosis and chronic obstructive pulmonary disease. PMID- 16719766 TI - TGF-beta: its role in asthma and therapeutic potential. AB - Asthma is a chronic disease of the airways affecting around 10% of the population. The majority of cases are well controlled with current therapies, however in approximately 20% of severe asthmatics the available therapeutic strategies are inadequate. Structural changes in the asthmatic airway, including an increase in smooth muscle mass and an increased deposition of extracellular matrix proteins, which correlate with airway hyperresponsivenes, reduced lung function and an increase in fibroblast/myofibroblast numbers, are not specifically targetted by current therapeutic agents and therefore represent an area of unmet need. The mechanisms involved in the development of airway remodelling are incompletely understood but are thought to involve one or more isoforms of transforming growth factor-beta (TGF-beta). The TGF-betas are pleiotropic mediators which have important roles in the regulation of inflammation, cell growth, differentiation and wound healing. All three mammalian isoforms of TGF-beta are present in the airways and at least TGF-beta1 and TGF beta2 have been shown to be increased in asthmatic airways and cells, together with evidence of increased TGF-beta signalling. In addition, evidence from animal models suggests that airway remodelling may be prevented or reversed using agents which target TGF-beta. Therefore modulation of TGF-betas or their activity represent a potential therapeutic target for asthma. This review focuses on the current knowledge of TGF-beta1-3, their their role in normal and asthmatic airways, as well as the potential for modulating the TGF-betas and their effects as a therapeutic approach to asthma. PMID- 16719767 TI - Extracellular matrix, integrins, and mesenchymal cell function in the airways. AB - Subepithelial fibrosis is one of the characteristic features of asthmatic airways. The fibrotic response includes an increase in volume occupied by extracellular matrix (ECM) tissue, and a change in the ECM composition favouring wound type collagens, fibronectin and a number of glycoproteins and proteoglycans normally associated with development. The altered ECM is likely to be deposited by the mesenchymal cells (including (myo) fibroblasts and smooth muscle) that are increased in number in asthmatic airways. In turn, the altered asthmatic ECM is likely to influence the function of the resident airway cells, and may be directly responsible for increasing proliferation, migration, ECM synthesis, inflammatory mediator release, and survival of resident mesenchymal cells. Therefore, the deposited ECM may perpetuate the disease phenotype. The different components of the ECM bi-directionally communicate with cells through a family of transmembrane receptors called integrins. Current research has begun to characterize: 1) the particular ECM components altered in airways disease; 2) the breadth of activity of different ECM components on airway cell function; and 3) the particular integrins responsible for mediating these effects. Further understanding of the role of integrins in transmitting responses of ECM in healthy or diseased airways may lead to novel targets for anti-asthma therapy. PMID- 16719768 TI - Role of chemokines in severe asthma. AB - The severe asthma phenotype is exhibited by a subset of asthma patients whose asthma symptom is poorly controlled by current therapies. Severe asthma represents a high unmet medical need and warrants research into the mechanisms driving the underlying pathophysiology. It is hypothesized that the underlying pathology associated with severe asthma is driving the symptoms experienced by these patients, which may share common features with mild to moderate asthma or may represent a unique pathological phenotype. For the purpose of this review, the pathophysiology associated with asthma in general are described and extended to incorporate severe asthma. Chemokines may contribute towards multiple features of asthma pathophysiology and this current review focuses on the biology of chemokines pertaining to asthma pathophysiology. Chemokines are important recruiters and activators of inflammatory cells and these infiltrating cells interact with resident cells, such as fibroblasts and it is through these pathways that chemokines appear to exert multiple biological actions. Clinical trials are underway with therapeutics targeting chemokine pathways for other inflammatory diseases. It is hoped that the information generated from these studies will contribute towards furthering our understanding of chemokine biology and be applied towards targeting severe asthma. PMID- 16719769 TI - Transcription factors in asthma: are transcription factors a new target for asthma therapy? AB - The essential features of persistent severe asthma include structural changes in the airway wall (remodelling). It is not known whether these are the sequelae of chronic inflammation or indeed its initiators. Several transcription factors have been implicated in the inflammatory process in asthma, including the glucocorticoid receptor (GR), NFkappaB, Activator Protein-1 (AP-1), Nuclear Factor of Activated T-cells (NF-AT), cyclic AMP Response Element Binding Protein and more recently, the CCAAT/Enhancer Binding Protein (C/EBP), Peroxisome Proliferator-activated Receptor (PPAR) and the bZIP transcription factor, Nrf2. Could a pathological de-regulation of one of these transcription factors explain the broad spectrum of asthma pathology and can their modulation lead to better symptom control? Although some of the transcription factors seem to be valid targets (NFkappaB, Nrf2 or STAT6) or tools (PPARgamma, -alpha and C/EBP-alpha) for new therapeutic approaches, since many transcription factors play a central role in tissue and organ homeostasis, a longterm general suppression or overexpression, would cause severe side effects in other organs. Cell type specific application of decoy or antisense oligonucleotides for NFkappaB, Nrf2 or STAT6, or specific agonists for PPARgamma and -alpha may help to control the inflammatory response in lung epithelial cells and infiltrated immune cells, but additional, unwanted, effects on other resident cells of the lung cannot be excluded and a beneficial effect over known anti-asthma drugs has first to be proven. In order to progress with such novel therapeutic strategies, the only option seems to be to link transcription factor inhibitors/activators to a cell type specific delivery system. PMID- 16719770 TI - Carbohydrates and epithelial repair - more than just post-translational modification. AB - Epithelia are the layers of cells that form barriers between external milieu and underlying tissues and thus, are important components of most organs of the body. Epithelial layers of organs, such as the lung, are exposed to various challenges resulting in frequent injury. Epithelial wound healing represents an important process by which repair restores the physical barrier lost as a result of cell damage and apoptosis. The repair of epithelial layers consists of a series of ordered events including epithelial cell spreading, migration proliferation and, differentiation. Carbohydrates attached to cell surface proteins and lipids can modulate the function of structures that they are conjugated to and therefore, can affect cell behavior. Although the basic mechanisms of epithelial repair are not entirely understood, many studies suggest glycoconjugates attached to proteins on the cell surface of epithelial cells play important roles in many of these cellular processes. In the present review, the role of carbohydrates in epithelial repair of different organs, including the sources of epithelial injury and current models of epithelial repair will be discussed with a focus on our understanding of the airway epithelium. With a better understanding of carbohydrates and their role in epithelial repair, new therapeutic targets for diseases involving damage to the epithelium can be identified. PMID- 16719771 TI - Zinc and its specific transporters as potential targets in airway disease. AB - The dietary group IIb metal zinc (Zn) plays essential housekeeping roles in cellular metabolism and gene expression. It regulates a number of cellular processes including mitosis, apoptosis, secretion and signal transduction as well as critical events in physiological processes as diverse as insulin release, T cell cytokine production, wound healing, vision and neurotransmission. Critical to these processes are the mechanisms that regulate Zn homeostasis in cells and tissues. The proteins that control Zn uptake and compartmentalization are rapidly being identified and characterized. Recently, the first images of sub-cellular pools of Zn in airway epithelium have been obtained. This review discusses what we currently know about Zn in the airways, both in the normal and inflamed states, and then considers how we might target Zn metabolism by developing strategies to monitor and manipulate airway Zn levels in airway disease. PMID- 16719772 TI - Protease-activated receptors (PARs) are partly pro-inflammatory and partly anti inflammatory: will PAR agonists or antagonists participate in future drug therapies? AB - Protease-activated receptors (PARs) are characterised by a unique mechanism of activation, which enables them to act as cellular sensors for protease activity. PARs are expressed throughout the cardiovascular, gastrointestinal and pulmonary systems, where they are potential drug targets for the treatment of disease. However, there are currently very few selective PAR antagonists or potent PAR agonists available as effective research tools, and moreover, there is considerable evidence to suggest that PARs can promote both pro-inflammatory and anti-inflammatory responses in a wide range of disease models. These confounding issues have, to date, prevented us from developing a clear understanding of the role of PARs in disease. Nevertheless, this review provides an overview of the distribution and function of PARs in the cardiovascular, gastrointestinal and pulmonary systems, and attempts to evaluate whether PAR agonists or antagonists have a place in future drug therapy. PMID- 16719773 TI - Rational drug design of selective epsilon opioid receptor agonist TAN-821 and antagonist TAN-1014. AB - Beta-endorphin (beta-EP) is generally classified as a mu and delta opioid receptor agonist but is also an agonist of the epsilon opioid receptor. Although several selective agonists and antagonists for mu, delta, and kappa opioid receptors are known, selective epsilon receptor agonists or antagonists have not been reported for some time. Recently, we designed and synthesized the selective epsilon receptor agonist, 17-(cyclopropylmethyl)-4,5alpha-epoxy-3,6beta-dihydroxy 6,14-endoethenomorphinan-7alpha-[N-methyl-N-phenethyl]carboxamide (TAN-821), and the selective epsilon receptor antagonist, 17-(cyclopropylmethyl)-4,5alpha-epoxy 6beta,21-epoxymethano-3-hydroxy-6,14-endoe-thenomorphinan-7alpha-(N phenethyl)carboxamide (TAN-1014). TAN-821 stimulated binding of the non hydrolyzable guanosine 5'-triphosphate analogue, guanosine 5'-(gamma-thio) triphosphate (GTPgammaS), to the mouse pons/medulla membrane via activation of the epsilon receptor. Moreover, TAN-821 given intracerebroventricularly (i.c.v.) produced marked, long-lasting, and dose-dependent antinociception in tail-flick and hot-plate tests. This antinociception induced by i.c.v. administered TAN-821 was blocked by i.c.v. pretreatment with the epsilon opioid receptor partial agonist beta-EP (1-27), but not the mu opioid receptor antagonist beta-FNA, the delta opioid receptor antagonist NTI, or the kappa opioid receptor antagonist nor BNI. On the other hand, i.c.v. injection of TAN-1014 alone produced no antinociception, and i.c.v. pretreatment with TAN-1014 attenuated the antinociception induced by i.c.v beta-EP. These results suggest that TAN-821 and TAN-1014 are respectively a selective epsilon receptor agonist and antagonist and that they may be useful tools for investigating the pharmacological properties of the epsilon opioid receptor. PMID- 16719774 TI - Chemistry and biology of chromatin remodeling agents: state of art and future perspectives of HDAC inhibitors. AB - Chromatin remodeling is a fundamental phenomenon in the life of eukaryotic cells, bearing implications to numerous physiological and pathological phenomena. This review outlines the chemistry of natural and synthetic agents endowed with the ability to interfere with such biological function, with a particular emphasis on histone deacetylase (HDAC) inhibitors. Other aspects covered in this article comprise structure activity relationships (SAR) and modes of action at molecular level, including the description of crystal structures of enzyme-inhibitor complexes. PMID- 16719775 TI - Roles of glycans and glycopeptides in immune system and immune-related diseases. AB - Almost all of the key molecules in organisms involved in the innate and adaptive immune response (including immunoglobulins, cytokines and cytokine receptors, complements, CD molecules, adhesions, T-cell receptors and major histocompatibility complex molecules) are glycoproteins. Besides, foreign antigens, such as many viral envelope proteins, are glycoproteins too. Carbohydrates attached to proteins or peptides are classified by the nature of their linkages to the protein, mostly as either N-linked (N-acetylglucosamine to asparagines) or O-linked (N-acetylgalactosamine to serine or threonine) oligosaccharides. The glycans have three major roles: firstly, the sugars confer stability on the proteins to which they are attached, protecting them from proteases and non-specific protein-protein interactions. Secondly, glycans play key roles in signal transduction, control of cell development and differentiation. Thirdly, specific regions of the oligosaccharide chains provide recognition epitopes, which influence innate and adaptive immune responses. Glycopeptides not only provide specific oligosaccharides, but also have specific information of amino acids sequences. The glycans and glycopeptides not only influence the structure and functions of immune molecules, but also influence the immune response. In addition, changes in glycans or glycopeptides may have a significant role in a variety of human immune-related diseases, such as rheumatoid, autoimmune disease, Wiskott-Aldrich syndrome, infection disease, cancer, etc. In this article, the roles of N-, O-glycans and glycopeptides in immune system and immune-related diseases are discussed. The potential therapeutic significance of the information is also mentioned. PMID- 16719776 TI - Targeting cytokines of the interleukin-12 family in autoimmunity. AB - In the past, autoimmunity was thought to be mediated by antibodies and immune complexes. It has now become clear that many autoimmune diseases, especially tissue specific, are T cell-mediated, or at least T cell-dependent. The pathogenesis of cell-mediated autoimmune diseases, including multiple sclerosis, uveitis, diabetes, arthritis, and others, is now thought to be, in a large measure, driven by interferon-gamma-producing, antigen-specific T cells, which are polarized toward the T helper type 1 (Th1) phenotype. Interleukin (IL)-12 and the more recently discovered IL-23 and IL-27 constitute a unique family of structurally-related, heterodimeric cytokines, which regulate cell-mediated immune responses and Th1-type inflammatory reactions. Thus, these cytokines may have a central role in the development and progression of cell-mediated autoimmune diseases. Therefore, pharmacologically targeting cytokines of the IL 12 family would be useful in the modulation of several autoimmune diseases. This review summarizes the recent findings concerning IL-12 family cytokine-mediated autoreactive inflammatory responses, and also describes some possible therapeutic interventions, including medicinal compounds at mitigating autoimmune inflammation. PMID- 16719777 TI - Aminolevulinic acid derivatives and liposome delivery as strategies for improving 5-aminolevulinic acid-mediated photodynamic therapy. AB - Photodynamic Therapy employing 5-aminolevulinic acid (ALA) as a precursor of the photosensitizer Protoporphyrin IX has become a promising approach to treat superficial cancers. However, the hydrophilic nature of the ALA molecule somewhat limits the penetration through the skin as well as all cell membranes. Different attempts are currently under investigation to enhance ALA penetration, such as the development of new synthetic and more lipophilic molecules derived from ALA and the incorporation of ALA into lipophilic vehicles such as liposomes. Among the new synthesized molecules, we can find ALA esters, ALA aminoacid derivatives and ALA dendrimers. In general, there is consensus that the promising results obtained in vitro with ALA esters cannot be reproduced in vivo. However, ALA methyl ester (1) has been widely used for treatment of skin malignancies and ALA hexyl ester (15) proved to be more powerful than ALA in bladder imaging. ALA aminoacid derivatives have been designed to use specific cellular aminopeptidases to targeting tumors, and it was shown that they can be metabolized to ALA with some specificity. PMID- 16719778 TI - Choline kinase: an important target for cancer. AB - Choline kinase (ChoK) is a cytosolic enzyme present in various tissues, which catalyzes the phosphorylation of choline to form phosphorylcholine (PCho) in the presence of ATP and magnesium. ChoK is important for the generation of two major membrane phospholipids, phosphatidylcholine (PC) and sphingomyelin (SM) and subsequently for the cell division. ChoK plays a vital role in cell signaling pathways and regulation of cell growth along with PCho involved in malignant transformation through ras oncogenes in different cancers such as breast, lung, colon, prostate, neuroblastoma, hepatic lymphomas, meningiomas and diverse murine tumours. The Ras effectors serine/threonine kinase (Raf-1), the Ral-GDP dissociation stimulator (Ral-GDS) and the phosphatidylinositol 3-kinase (PI3K) are involved in the activation of ChoK during tumorigenesis. ChoK gene induction seems to be associated with certain cell stress or cell defense. Nowadays, RNAi appear to be one of the most promising routes in the cancer therapy. The anticancer potential of both stable expression of siRNAs and their high sequence specificity by RNAi mediated suppression of oncogenic ras in human pancreatic carcinoma, human melanomas and ovarian cancer has been observed. It has an important role in sequence specific post-transcriptional gene silencing mechanism. Presently, the crystal structure of Caenorhabditis elegans choline kinase A-2 (ChoKA-2) is available, which may be useful for comparative modeling of human ChoK and further modeling studies. The present review aims at the general overview of importance, expression, structure, progress in molecular modeling, active site analysis and inhibitors of ChoK. It also highlights the recent role of ChoK in various types of Ras-dependent and Ras-independent carcinogenesis. PMID- 16719779 TI - Reactive oxygen species-induced gastric ulceration: protection by melatonin. AB - Gastric hyperacidity and ulceration of the stomach mucosa due to various factors are serious health problems of global concern. Although the mechanism of acid secretion from the parietal cells is now well understood, the processes involved in gastric ulceration are still not clear. Among the various causes of gastric ulceration, lesions caused by stress, alcohol consumption, Helicobacter pylori and due to use of non-steroidal anti-inflammatory drugs have been shown to be mediated largely through the generation of reactive oxygen species, especially the hydroxyl radical. A number of excellent drugs, developed over the decades, have proven useful in controlling hyperacidity and ulceration although their long term use is reported to be associated with various side effects. Hence the investigations continue with an objective to find a compound possessing anti secretory, anti-ulcer and antioxidant properties which will serve as a therapeutic agent to reduce gastric hyperacidity and ulcers. This article describes the role of reactive oxygen species in gastric ulceration, briefly presents a note on the currently available drugs controlling them, and focuses on the role of melatonin, a pineal secretory product, in protecting against gastric lesions. In experimental studies, melatonin has been shown to be effective in reducing mucosal breakdown and ulcer formation in a wide variety of situations. Additionally, the low toxicity of melatonin supports further investigation of this molecule as a promising gastro-protective agent. Finally, we include a commentary on how melatonin research with respect to gastric pathophysiology can move forward with a view to eventually using this indole as a therapeutic agent alone or in combination with the existing drugs to control gastric ulceration in humans in order to increase their efficacy and/or to reduce their side effects. PMID- 16719780 TI - Antidiabetic agents from medicinal plants. AB - Currently available therapeutic options for non-insulin-dependent diabetes mellitus, such as dietary modification, oral hypoglycemics, and insulin, have limitations of their own. Many natural products and herbal medicines have been recommended for the treatment of diabetes. The present paper reviews medicinal plants that have shown experimental or clinical antidiabetic activity and that have been used in traditional systems of medicine; the review also covers natural products (active natural components and crude extracts) isolated from the medicinal plants and reported during 2001 to 2005. Many kinds of natural products, such as terpenoids, alkaloids, flavonoids, phenolics, and some others, have shown antidiabetic potential. Particularly, schulzeines A, B, and C, radicamines A and B, 2,5-imino-1,2,5-trideoxy-L-glucitol, beta homofuconojirimycin, myrciacitrin IV, dehydrotrametenolic acid, corosolic acid (Glucosol), 4-(alpha-rhamnopyranosyl)ellagic acid, and 1,2,3,4,6 pentagalloylglucose have shown significant antidiabetic activities. Among active medicinal herbs, Momordica charantia L. (Cucurbitaceae), Pterocarpus marsupium Roxb. (Leguminoceae), and Trigonella foenum graecum L. (Leguminosae) have been reported as beneficial for treatment of type 2 diabetes. PMID- 16719781 TI - The mercaptopyruvate pathway in cysteine catabolism: a physiologic role and related disease of the multifunctional 3-mercaptopyruvate sulfurtransferase. AB - The conversion of cysteine to 3-sulfino-alanine is a major pathway in cysteine catabolism. Cysteine dioxygenase catalyzes the reaction and dietary intake of the essential amino acid methionine and the semi-essential amino acid cysteine increases the level of this enzyme by suppressing enzyme degradation via polyubiquitination. The production of cellular antioxidants such as glutathione, thioredoxin, and their families is important in cysteine metabolism, and these cellular antioxidants have critical roles in the maintenance of the cellular redox status. The mercaptopyruvate pathway, in which cysteine or aspartate transaminase catalyzes the transamination from cysteine to 3-mercaptopyruvate and then 3-mercaptopyruvate sulfurtransferase catalyzes the transsulfuration from 3 mercaptopyruvate to pyruvate, also contributes to maintain the cellular redox. 3 Mercaptopyruvate sulfurtransferase serves as an antioxidant protein: when the enzyme is exposed to stoichiometric amounts of the oxidant hydrogen peroxide, it is inhibited via the formation of low redox sulfenate at the catalytic site cysteine. On the other hand, activity is restored by the reductant dithiothreitol or reduced thioredoxin. 3-Mercaptopyruvate sulfurtransferase also detoxifies cyanide via transsulfuration from a stable persulfide at the catalytic site cysteine, a reaction intermediate, suggesting that cyanide detoxification is not necessarily an enzymatic reaction. Furthermore, a congenital defect of the enzyme causes mercaptolactate-cysteine disulfiduria associated with or without mental retardation, although the pathogenesis remains unclear. These facts suggest that 3-mercaptopyruvate sulfurtransferase has physiologic roles as an antioxidant and a cyanide antidote; is essential for neural function, and participates in cysteine degradation. PMID- 16719782 TI - Lipid bilayer fragments and disks in drug delivery. AB - Certain lipids can be dispersed as colloidally stable bilayer fragments (BF) or disks with interesting properties for solubilization and delivery of hydrophobic or amphiphilic drugs. They were first observed and characterized as such in the nineties but remained silent in the literature regarding applications for drug delivery. Only recently their potential for delivery of hydrophobic drugs started to be realized. This review deals with electrostatically or sterically stabilized bilayer fragments which provided excellent solubilization sites for antifungal drugs, acted as drugs themselves against bacteria or fungus, could be loaded with amphiphilic drugs or produced lipid-covered drug particles to be delivered as a synergistic formulation in vivo. PMID- 16719783 TI - Protein transduction: cell penetrating peptides and their therapeutic applications. AB - Cell penetrating proteins or peptides (CPPs) have the ability to cross the plasma membranes of mammalian cells in an apparently energy- and receptor-independent fashion. Although there is much debate over the mechanism by which this "protein transduction" occurs, the ability of CPPs to translocate rapidly into cells is being exploited to deliver a broad range of therapeutics including proteins, DNA, antibodies, oligonucleotides, imaging agents and liposomes in a variety of situations and biological systems. The current review looks at the delivery of many such molecules by various CPPs, and their potential therapeutic application in a wide range of areas. CPP ability to deliver different cargoes in a relatively efficient and non-invasive manner has implications as far reaching as drug delivery, gene transfer, DNA vaccination and beyond. Although many questions remain to be answered and limitations on the use of CPPs exist, it is clear that this emerging technology has much to offer in a clinical setting. PMID- 16719784 TI - Intervention of Toll-like receptor-mediated human innate immunity and inflammation by synthetic compounds and naturally occurring products. AB - Human innate immunity plays a pivotal role in host defense against various microbial challenges. Mediated by a family of Toll-like-receptors (TLR) and associated intracellular downstream signaling molecules, human innate immunity can specifically recognize diverse microbial products and many other non microbial environmental cues. Beyond its role of providing first line of defense, activation of innate immunity signaling can lead to expression of diverse pro- and anti- inflammatory mediators, which are critical for regulating various cell and tissue metabolism. Alteration in innate immunity signaling may therefore lead to infection and inflammatory diseases such as atherosclerosis, diabetes, and cancer. TLR receptors as well as intracellular signaling proteins can serve as therapeutic targets for treating various inflammatory diseases. Several synthetic ligands of TLR receptors such as lipid A analogs, poly(I:C), loxoribine, oligodeoxynucleotides have been shown to be effective in regulating innate immune response. This review discusses the potential, challenge, and recent progress of developing synthetic as well as naturally occurring TLR ligands in regulating innate immunity and treating inflammatory diseases. PMID- 16719785 TI - Premature ovarian failure (POF) syndrome: towards the molecular clinical analysis of its genetic complexity. AB - The Premature Ovarian Failure (POF) syndrome is a very heterogeneous clinical disorder due probably to the complex genetic networks controlling human folliculogenesis. Clinical subgroups of POF patients whose aetiology of ovarian failure is based on the same genetic factors are therefore difficult to establish. Some experimental evidence suggests that these genes might be clustered on the female sex chromosome in the POF1 and POF2 loci. This review is aimed to present an overview of the actual structural changes of the X chromosome causing POF, and to present a number of X and autosomal female fertility genes which are probably key genes in human folliculogenesis and are therefore prominent POF candidate genes. Towards the molecular analysis of their functional contribution to the genetic aetiology of POF in the clinic, an interdisciplinary scheme for their diagnostic analysis is presented in a pilot study focussed on chromosome analyses and the expression analysis of some major POF candidate genes (DAZL, DBX, FOXL2, INHalpha, GDF9, USP9X) in the leukocytes of 101 POF patients. It starts with a comprehensive and significantly improved clinical diagnostic program for this large and heterogeneous patient group. PMID- 16719786 TI - Synthesis, biological evaluation and 3D-QSAR of 1,3,5-trisubstituted-4,5-dihydro (1H)-pyrazole derivatives as potent and highly selective monoamine oxidase A inhibitors. AB - The present report provides a extended study of the chemistry, the inhibitory activity against monoamino oxidases (MAO), and molecular modeling including the 3D-QSAR hypothesis of 1,3,5-trisubstituted-4,5-dihydro-(1H)-pyrazole derivatives. Four series of about eighty novel pyrazoline derivatives were prepared and investigated for their ability to inhibit the activity of the A and B isoforms of MAO selectively. Most of the new synthesized compounds proved more reversible, potent, and selective inhibitors of MAO-A than of MAO-B, and could be taken into account to develop the search further in this field, knowing that reversible and selective MAO-A inhibitors are used as antidepressant and antianxiety drug. The 30 most active compounds show inhibitory activity on MAO-A in the 8.6 x 10(-8) - 9.0 x 10(-9)M range. Moreover, it should be pointed out that for most of them a high IC(50) > or = 10(-9)M value is associated with a high A-selectivity (Selectivity Index MAO-B/MAO-A in the 10,000-16,250 range). Furthermore, due to the presence of a chiral centre at the C5 position of the pyrazole moiety, we performed the semi-preparative chromatographic enantioseparation of the most potent, selective, and chiral compounds. The separated enantiomers were then submitted to in vitro biological evaluation, and from the results of these experiments it has been possible to point out a difference in inhibiting the two isoforms selectively between the racemic mixture and the single enantiomers. The molecular modeling work was carried out combining the Glide docking approach with CoMFA with the aim to rationalize the structure-activity relationships of each pyrazoline inhibitor toward MAO-A and MAO-B isoforms and to derive a suitable selectivity model. PMID- 16719787 TI - Antiobesity therapy: emerging drugs and targets. AB - Obesity and its associated morbidities and mortalities are the effects of imbalance between energy intake and expenditure. The healthcare burden for the treatment of obesity is significantly high, due to increased risk of secondary chronic diseases such as hypertension and associated co-morbidities such as diabetes and cardiovascular disease. Lack of physical activity, high fat diets and sedentary life styles are major factors contributing to obesity. However, genetic predisposition and ethnicity are increasingly found to cause obesity. Till date, approved therapeutics have addressed excess energy intake by acting on central neural circuits that regulate feeding or on peripheral mechanisms to reduce nutrient absorption from the gut. These approaches have met with moderate success; and recently with safety concerns, leaving an unmet medical need for effective and safe pharmacotherapy for obesity thereby posing a significant challenge to pharmaceutical industry. Potential antiobesity drugs, which are being investigated by different companies, can be classified in 4 broad categories: 1) Agents that primarily decrease appetite through central action; 2) Agents that primarily increase metabolic rate or affect metabolism through peripheral action; 3) Agents that act on gastrointestinal tract; and 4) Agents that not only affect obesity but also overall Metabolic Syndrome. The current review will deal mainly with different molecules, which are under development for the above-mentioned targets and also their potential benefits and disadvantages. PMID- 16719788 TI - Advance in antitumor agents targeting glutathione-S-transferase. AB - Glutathione-S-transferases (GSTs; EC 2.5.1.18), a family of phase II detoxification enzymes, catalyze the conjugation of glutathione with broad substrates including chemotherapeutic agents, and are involved in cell protection against apoptotic signals by inhibiting the stress-signaling cascade mediated by ASK1 (Apoptosis signal-regulating kinase)-JNK (c-Jun N-terminal kinase). As GSTs are overexpressed in some malignant tumors, GSTs is a promising therapeutic target. This article reviews the progress in the development of GSTs inhibitors and GSTs activated prodrugs. PMID- 16719790 TI - Compromise and care of the brain's compartments: the quintessence of the neurovascular unit. PMID- 16719789 TI - Intrabody-based approaches to cancer therapy: status and prospects. AB - Continuing developments from the study of cancer at the molecular level are yielding increasing numbers of targets that may be used for therapeutic intervention. Advances in the field of antibody engineering over the past several decades have given scientists the capability of directing the highly specific interaction of antibodies with antigens inward, to the intracellular compartments of living cells. These intracellular antibodies, i.e., intrabodies, are being developed to bind to, neutralize, or modify the function or localization of cancer-related targets and thereby affect the malignant phenotype. This has resulted in a promising new tool for the study and treatment of cancer. Due to recent advances in the development of the antibody engineering technologies, increasing numbers of intrabodies are being exploited to a growing list of cancer related, as well as other disease targets. There are still, however, many technical issues, particularly related to clinical applications of the intrabodies, that must be addressed before the full promise of this class of therapeutic agent is realized. This review will focus on the recent progress in the generation and use of intrabodies in the field of oncology. The technical issues associated with their further development will also be discussed. PMID- 16719791 TI - Endothelin-1 impairs retrograde axonal transport and leads to axonal injury in rat optic nerve. AB - The purpose of this study was to examine the effects of endothelin-1 (ET-1) on retrograde axonal transport in the rat optic nerve. Vehicle or ET-1 (0.2, 1, or 5 pmol/eye) were injected into the vitreous body in Sprague-Dawley rats. Retinal vessels were observed, using a fundus camera, before, and at 10 min, 3 days and 7 days after a single intravitreous injection. Two days after the injection, a neuronal tracer, fluoro gold, was administered via the superior colliculi to retrogradely label active retinal ganglion cells (RGCs). Five days after the tracer administration, retrogradely labeled RGCs were evaluated in the flat mounted retina, and cross sections from each optic nerve were graded for injury by four independent, masked observers. ET-1 at 5 pmol/eye caused a significant constriction of retinal vessels (versus the vehicle-treated group) at 10 min after the injection. Intravitreous injection of ET-1 caused a dose-related decrease in the number of retrogradely labeled RGCs. Injection of 5 pmol/eye ET-1 led to a statistically significant decrease in the number of retrogradely labeled RGCs (versus the vehicle-treated group). ET-1 at 1 and 5 pmol/eye caused histological optic nerve damage (evaluated using a graded scale). The histological optic nerve damage correlated with the number of retrogradely labeled RGCs. In conclusion, a single intravitreous injection of ET-1 impaired retrograde axonal transport in the rat optic nerve and this impairment correlated with the histological optic nerve damage. PMID- 16719792 TI - Sigma receptor activation reduces infarct size at 24 hours after permanent middle cerebral artery occlusion in rats. AB - The only available treatment for embolic stroke is recombinant tissue plasminogen activator, which must be administered within three hours of stroke onset. We examined the effects of 1,3-di-o-tolyguanidine (DTG), a high affinity sigma receptor agonist, as a potential treatment for decreasing infarct area at delayed time points. Rats were subjected to permanent embolic middle cerebral artery occlusion (MCAO) and allowed to recover before receiving subcutaneous injections of 15 mg/kg of DTG at 24, 48, and 72 hours. At 96 hours the rats were euthanized, and brains harvested and sectioned. Infarct areas were quantified at the level of the cortical/striatal and cortical/hippocampal regions in control (MCAO-only) and DTG treated animals using a marker for neurodegeneration, Fluoro-Jade. DTG treatment significantly reduced infarct area in both cortical/striatal and cortical/hippocampal regions by >80%, relative to control rats. These findings were confirmed by immunohistochemical experiments using the neuronal marker, mouse anti-neuronal nuclei monoclonal antibody (NeuN), which showed that application of DTG significantly increased the number of viable neurons in these regions. Furthermore, DTG blocked the inflammatory response evoked by MCAO, as indicated by decreases in the number of reactive astrocytes and activated microglia/macrophages detected by immunostaining for glial fibrillary acidic protein (GFAP) and binding of isolectin IB4, respectively. Thus, our results demonstrate that the sigma receptor-selective agonist, DTG, can enhance neuronal survival when administered 24 hr after an ischemic stroke. In addition, the efficacy of sigma receptors for stroke treatment at delayed time points is likely the result of combined neuroprotective and anti-inflammatory properties of these receptors. PMID- 16719793 TI - Endothelial colony forming capacity is related to C-reactive protein levels in healthy subjects. AB - The majority of clinical studies on endothelial progenitor cells (EPCs) focuses on the role of these cells in cardiovascular diseases and no systematic studies exist regarding their variations in healthy subjects. In order to define the burden of angiogenesis in physiological conditions we assessed the frequency of peripheral blood endothelial colonies (PB-ECs) and their relation with other factors possibly involved in their function such as high-sensitivity C-reactive protein (hs-CRP), endothelial cell-specific mitogen factor (VEGF) and tissue inhibitor of metalloproteinases-1 (TIMP-1) in a highly selected healthy population. A PB sample was obtained from 37/47 healthy subjects (age 40.2+/ 15.0yrs; M/F 15/22) without known cardiovascular risk factors. The serum level of hs-CRP, VEGF, TIMP-1, the frequency of PB-ECs by clonogenic assay, and the number of early EPCs and late EPCs by flow cytometry analysis were evaluated. PB-ECs were formed by 40.5% of studied subjects with a mean of 0.40+/-0.82 colonies/10(6) cells. The differences in the frequency of colony formation between genders were not statistically significant. The subjects with PB-ECs were characterized by higher values of hs-CRP, when compared with those not forming colonies, 0.276+/-0.230 vs 0.095+/-0.077 mg/l (p=0.003) respectively, and of VEGF, 328.3+/-162.9 vs 202.68+/-118.53 pg/ml (p=0.02). No significant differences were found in TIMP-1 values. The EPC clonogenic potential seems to be related to hs-CRP and VEGF levels even in healthy population supporting the concept that these mediators are involved in physiological ECs function. PMID- 16719795 TI - Cerebral ischemia and angiogenesis. AB - Angiogenesis occurs in a wide range of conditions. As ischemic tissue usually depends on collateral blood flow from newly produced vessels, acceleration of angiogenesis should be of therapeutic value to ischemic disorders. Indeed, therapeutic angiogenesis reduced tissue injury in myocardial or limb ischemia. In ischemic stroke, on the other hand, angiogenic factors often increase vascular permeability and thus may deteriorate tissue damage. In order to apply safely the therapeutic angiogenesis for ischemic stroke treatment, elucidating precise mechanism of brain angiogenesis is mandatory. In the present article, we review previous reports which investigated molecular mechanisms of angiogenesis. Endothelial cell mitogens, enzymes that degrade surrounding extracellular matrix, and molecules implicated in endothelial cells migration are induced rapidly in the ischemic brain. Their possible neuroprotective or injury exacerbating effects are discussed. Because therapeutic potential of angiogenic factors application had gained much attention, we here extensively reviewed relevant previous reports. In the future however, there is a need to consider angiogenesis in relation with regenerative medicine, as angiogenic factors sometimes possess neuron producing property. PMID- 16719796 TI - Neurovascular mechanisms of hypertension in pregnancy. AB - Normal pregnancy is associated with significant changes in the neuronal and vascular control mechanisms of blood pressure (BP). Preeclampsia (PE) is a major complication of pregnancy characterized by proteinuria, and increased vascular resistance and BP. If untreated, PE leads to eclampsia with serious seizures and severe hypertension. However, the neurovascular mechanisms of hypertension in pregnancy and PE are unclear. Studies in animal models of hypertension in pregnancy suggest that inadequate cytotrophoblast invasion of uterine spiral arteries causes reduction in uteroplacental perfusion pressure leading to placental ischemia/hypoxia. Placental ischemia may promote the release of biologically active factors such as cytokines and reactive oxygen species. These circulating factors may increase the vascular permeability, cross the blood-brain barrier, and affect the sympathetic tone and the neuronal control mechanisms of BP. Placental factors could also cause endothelial cell dysfunction and inhibit nitric oxide (NO)-cyclic guanosine monophosphate (cGMP), prostacyclin (PGI(2)) cyclic adenosine monophosphate (cAMP), and hyperpolarizing factor vascular relaxation pathways. Additionally, placental factors may induce endothelium derived contracting factors such as endothelin, thromboxane and angiotensin II, which stimulate Ca(2+)-dependent vascular smooth muscle (VSM) contraction or increase protein kinase C activity and enhance myofilament sensitivity to intracellular free calcium concentration ([Ca(2+)](i)). The increased sympathetic tone combined with systemic decrease in endothelium-dependent vascular relaxation and enhanced VSM contraction may contribute to the increased vascular resistance and BP associated with PE. The hypertensive state in severe PE may weaken the blood-brain barrier and precipitate convulsions and cerebral hemorrhage. Careful monitoring of maternal neuronal, endothelial, and VSM function during pregnancy should circumvent the life-threatening neurovascular complications of PE eclampsia. PMID- 16719797 TI - Autoantibodies associated with psychiatric disorders. AB - Growing evidence suggests that autoantibodies to neuronal or endothelial targets in psychiatric disorders exist and may be pathogenic. This review describes and discusses the possible role of autoantibodies related to the psychiatric manifestations in autoimmune diseases, autoantibodies related to the psychiatric disorders present in post-streptococcal diseases, celiac disease, chronic fatigue syndrome and substance abuse, and autoantibodies related to schizophrenia and autism, disorders now considered of autoimmune origin. PMID- 16719794 TI - Group I metabotropic receptor neuroprotection requires Akt and its substrates that govern FOXO3a, Bim, and beta-catenin during oxidative stress. AB - Metabotropic glutamate receptors are expressed throughout the nervous system, but their function as well as their ability to promote neuronal survival rests heavily upon the intracellular mechanisms governed by this family of G-proteins. In this regard, we examined one of the primary pathways that can oversee cell survival, namely protein kinase B (Akt1), and its functional integration with some of its substrates that may work in concert with group I metabotropic glutamate receptor (mGluRI) activation to protect primary hippocampal neurons during oxidative stress. We demonstrate that neuroprotection against free radical injury through mGluRI activation with DHPG requires the activation of Akt1, since loss of Akt1 activity assessed through its GSK-3alpha/beta substrate by pharmacological blockade of the phosphatidylinositide-3-kinase pathway or the gene silencing of Akt1 expression prevents neuronal protection during mGluRI activation. Closely coupled to the robust neuroprotection by mGluRI activation are the inhibitory phosphorylation and prevention of caspase 3 cleavage of the Forkhead transcription factor FOXO3a, the down-regulation of Bim expression, and the protection of beta-catenin by Akt1 against phosphorylation and degradation to promote its translocation from the cytoplasm to the nucleus and allow it to assist with a "pro-survival" cellular program. Further insight into the cellular mechanisms that determine neuronal protection by the metabotropic glutamate system will foster the successful therapeutic development of mGluRs for neurodegenerative disorders. PMID- 16719798 TI - Vascular changes of the retina and choroid in systemic lupus erythematosus: pathology and pathogenesis. AB - Systemic lupus erythematosus (SLE) is a chronic autoimmune disorder that affects multiple organ systems. When eyes are involved, the principle manifestations are hemorrhage, retinal cotton wool spots, microangiopathy and vaso-occlusion. Research in the past two decades has significantly contributed to our understanding about this disease in general and its therapeutic management, although knowledge about the mechanism of ocular involvement and pathogenesis in SLE is limited. This is an important issue, because the ocular symptoms in this disease could be potentially sight threatening in acute cases. Here, we present an overview of the clinical and histopathologic features of retinal and choroidal vascular changes, as seen in patients with SLE. We discuss the role of immune complex deposition in vascular pathogenesis in the eye. Reports indicated an involvement of antiphospholipid antibodies (APAs) in the retinal and choroidal vasculopathy in SLE, although their precise role in this process is uncertain. It is important to look for mechanisms of immune complex-mediated vasculopathy and role of inflammatory mediators in this process in SLE. For this, established animal models can be utilized in research to learn about the precise role of various autoantibodies and complements involved in disease pathogenesis. A clear knowledge about the immunopathogenesis is warranted, and the rationale for the future therapy should be based on reducing vascular inflammation as well as ameliorating autoimmunity in this disease. PMID- 16719799 TI - Successful drug discovery. PMID- 16719800 TI - Discovery and clinical development of dutasteride, a potent dual 5alpha-reductase inhibitor. AB - In this review the preclinical medicinal chemistry, biochemistry and clinical results achieved in the treatment of prostatic disease with dutasteride, a dual inhibitor of type 1 and type 2,5alpha-reductase are described. During the discovery phase, dutasteride was optimized to inhibit both forms of human 5 alpha reductase (5AR) via extensive structure activity relationship studies versus the cloned human isozymes. Dutasteride has subsequently been shown to improve disease measures in patients with symptomatic benign prostatic hyperplasia (BPH) in three randomized, placebo-controlled, Phase III clinical studies lasting for 2 years. Additionally, dutasteride is now under study for the ability to reduce the incidence of prostate cancer in men at high risk of the disease--an indication where the unique dual inhibitor nature, half-life and tolerability of dutasteride may be especially significant factors in determining treatment success. The connections between preclinical drug design and clinical outcomes during the discovery and development of dutasteride are exemplified. PMID- 16719801 TI - Optimization of small molecule drugs binding to highly polar target sites: lessons from the discovery and development of neuraminidase inhibitors. AB - Binding affinity optimization of small molecules interacting with polar binding sites on target proteins is a formidable, but not uncommon challenge in drug discovery. The challenge relates to the difficulty of integrating favourable and unfavourable polar, non-polar and conformation contributions into overall favourable binding energies. This review describes the surprising breakthrough findings leading to the development of Tamiflu, a clinically efficacious orally bioavailable drug targeting the active site of influenza neuraminidase (NA). The NA active site is highly polar and formed mostly by arginine, aspartate and glutamate residues. This active site structure evolved for efficient interaction with charged sialic acid moieties on glycoproteins and stabilization of an oxocarbonium ion in the transition state of the neuraminidase reaction. The initial strategy of optimizing polar interactions in transition state analogs led to NA inhibitors (NAIs) with sub-nanomolar binding affinities, but such compounds were highly polar and lacked oral bioavailability. The realization of the possibility to achieve high affinity binding in a highly polar active site through optimization of non-polar and van-der-Waals interactions initially appeared counterintuitive and required a few serendipitous findings, but was key to reduce the polarity of drug candidates, avoid large desolvation penalties and achieve oral bioavailability. PMID- 16719802 TI - Lessons from the drug discovery of lapatinib, a dual ErbB1/2 tyrosine kinase inhibitor. AB - Protein Kinases offer many opportunities for drug intervention points since phosphorylation is the most common post-translational modification. Phosphorylation regulates activity, location, degradation, conformation and the aberrant activity is implicated in many diseases, including cancer, inflammation, cardiovascular and central nervous system diseases. The focus of this review will be on the generation of highly effective signaling inhibitors targeting members of the erbB family of receptor tyrosine kinases, EGFR and ErbB-2, also known as transmembrane Type 1 receptor tyrosine kinases of the HER family of receptors. Ligand binding to the receptor causes a conformational change which activates the tyrosine kinase domain leading to autophosphorylation. This autophosphorylation activates the RAS/mitogen activated protein (MAP) kinase and phosphoinositol-3 kinase (PI3K) pathways leading to a myriad of signaling and cellular activities. Type 1 receptors are over-expressed in a variety of cancers and generally correlate with poor prognosis. For this reason, scientists at GlaxoSmithKline and many others in the scientific community, target the ATP binding site of the intracellular portion of the protein to block the aberrant signaling event. This review intends to cover the lessons learned in the discovery of lapatinib (GW572016, GW2016) by pulling together the various different publications that have been generated in distinct disciplines on aspects of the drug discovery program. Data analyses and correlation of assay data to help with the design of drug like molecules will be included and will demonstrate a break from the traditional focus on absolute potency as a guiding factor in lead compound selection. PMID- 16719803 TI - Biochemical mechanisms of New Molecular Entities (NMEs) approved by United States FDA during 2001-2004: mechanisms leading to optimal efficacy and safety. AB - The United States FDA approved 85 New Molecular Entities (NMEs) during the period from January 2001 to November 2004 of which 60 were pharmaceuticals with known molecular targets. The majority targeted enzymes (48%) or G-protein coupled receptors (GPCRs) (33%). Eighty percent of the NMEs interacted at the same site as endogenous effector; either as competitive inhibitor/antagonist (67%) or agonist (13%). Three biochemical operations defined the modes of action of the NMEs: 1) mass action competition (equilibrium), 2) a drug stabilized conformational change in the target that is important to the response (conformational) and/or 3) drug action is less-responsive to mass action competition with effectors due to non-equilibrium kinetics (non-equilibrium kinetic). Approximately 80% of the NMEs elicit a response utilizing conformational and/or non-equilibrium kinetic mechanisms. The remaining 20% of NMEs find mass action competition with the endogenous substrate or ligand sufficient for therapeutic utility. These observations indicate that for the majority of drug targets, mass action driven equilibrium binding alone may not be sufficient for maximal therapeutic utility. A key determinant of the biochemical mode of action for these NMEs was to minimize the potential for toxicity, either by providing a maximal response at a low dose to minimize off-target toxicities, or by providing a mechanism to minimize the incidence of mechanism-based toxicity while retaining a sufficiently efficacious response. This principle appears to be independent of target class and provides insight as to intrinsic biochemical features and approaches required for a maximal therapeutic index. PMID- 16719804 TI - A medicinal chemistry perspective on 4-aminoquinoline antimalarial drugs. AB - A broad overview is presented describing the current knowledge and the ongoing research concerning the 4-aminoquinolines (4AQ) as chemotherapeutic antimalarial agents. Included are discussions of mechanism of action, structure activity relationships (SAR), chemistry, metabolism and toxicity and parasite resistance mechanisms. In discussions of SAR, particular emphasis has been given to activity versus chloroquine resistant strains of Plasmodium falciparum. Promising new lead compounds undergoing development are described and an overview of physicochemical properties of chloroquine and amodiaquine analogues is also included. PMID- 16719805 TI - From artemisinin to new artemisinin antimalarials: biosynthesis, extraction, old and new derivatives, stereochemistry and medicinal chemistry requirements. AB - The artemisinin derivatives, dihydroartemisinin (DHA), artesunate, atemether and arteether, are currently used for treatment of malaria in artemisinin combination therapies (ACT) with longer half-life drugs. The demand is enormous--in 2005, the estimated global demand for one such ACT alone, artemether-lumifantrine, which constitutes about 70% of all current clinically-used ACTs, is for 120 million adult treatment courses. At 0.5 gm of artemether per total dose regimen, the amount of artemisinin required is approximately 114 tons. This has placed substantial stress on total artemisinin supplies world-wide, and considerable attention is being focussed on enhancing availability of artemisinin by improvement in horticultural practice and extraction of artemisinin from Artemisia annua. Artemisinic acid, which also occurs in A. annua, can be converted into artemisinin and is the ultimate target of a biotechnological approach, which if successful, will augment artemisinin supply in the future. The conversion of artemisinin into the known artemisinin derivatives, and problems with the methods are critically reviewed. Some attention is paid to mechanistic aspects which clarify stereochemistry. The current artemisinins are by no means ideal drugs. Artesunate in particular is incompatible with basic quinolines by virtue of proton transfer, and has intrinsic chemical instability. At pH 1.2, conversion to DHA is rapid, with t(1/2) 26 min, and at pH 7.4, t(1/2) is about 10 hours. With a pK(a) of 4.6, over 99% of artesunate will be ionized at pH 7.4, and thus uptake by passive diffusion from the intestinal tract will be minimal. Although a considerable effort has been vested in the search for new artemisinins, largely through functionalization of artemisinin at C-10, O-11 or at C-15 via artemisitene, or of DHA at C-10, deliberate enhancement of the 'druggability' of artemisinins by reducing lipophilicity, which at the same time will attenuate the neurotoxicity characteristic of the current derivatives, and enhance absorption, by and large has not been considered. A review of the various types of newer derivatives is given together with a consideration of medicinal chemistry aspects. PMID- 16719806 TI - Mechanisms of drug action and drug resistance in Leishmania as basis for therapeutic target identification and design of antileishmanial modulators. AB - The control of Leishmania infections relies primarily on chemotherapy. The arsenal of drugs available against Leishmania infections is limited and includes pentavalent antimonials, pentamidine, amphotericin B, miltefosine, paromomycin, allopurinol, and few other drugs at various stages of their development process. Knowledge about action and resistance mechanisms involved may allow the development of new drugs that minimise or circumvent drug resistance or may identify new targets for drug development. The aim of this review is to propose some chemical topics to design new modulators from the mechanisms of action of drugs and resistance mechanisms to drugs used in the clinic against Leishmania infections. Thus, different classes of ABC transporters extrude antimonials in Leishmania resulting in drug-resistant phenotypes. Compounds interfering with thiol and polyamine metabolism could be designed to inhibit the antimonial detoxication and therefore, such compounds could be used in combination with antimonials. New diamidines could be synthesized in regard to their ability to inhibit topoisomerase II. The challenge for amphotericin B is to be absorbed by oral route requiring labile physico-chemical modifications. New sesquiterpens and flavonoids have to be developed as reversant of antimonial resistance. Although some studies have focused on developing inhibitors against these resistant phenotypes, new efficient modulators that are able to inhibit drug efflux are needed. PMID- 16719807 TI - Glycine/NMDA receptor antagonists as potential CNS therapeutic agents: ACEA-1021 and related compounds. AB - Glutamate is the main excitatory neurotransmitter in central nervous system (CNS) and NMDA receptors are one of the major classes of ionotropic glutamate receptors. NMDA receptors have been known to play critical roles in normal CNS activities, as well as in many pathological conditions, including both acute and chronic diseases. The discovery of glycine as a coagonist of NMDA receptors has led to intensive research of glycine/NMDA antagonists as potential CNS drugs. The robust efficacy of glycine/NMDA antagonists, such as ACEA-1021 (5), in animal model of brain ischemia, together with good safety profile in animal models and in clinical trials, suggested that this class of NMDA antagonists should have good chance of success in the clinic as neuroprotectants. The clinical trial of ACEA-1021 for stroke was discontinued, mainly due to low solubility and lack of metabolism of the drug that led to the observation of crystals in the urine of some of the patients. However, through SAR studies, compounds such as ACEA-1416 (10) have been identified with improved properties, such as higher in vivo potency and site for potential metabolism. Therefore these compounds should be able to overcome some of the liabilities of ACEA-1021 and potentially could be developed as neuroprotectants. Based on the preclinical and clinical studies of glycine/NMDA antagonists, as well as the clinical experiences with t-PA, initiation of treatment within a short time window after the onset of stroke could be critical for the success of these antagonists in clinical trials. This can be accomplished by implementing the procedure developed for t-PA clinical trials, with modification based on the safety profile of glycine/NMDA antagonists, for future clinical trial to administer the drug as soon as possible after stroke onset. In addition, glycine/NMDA antagonists also have other potential therapeutic applications, such as for the treatment of traumatic brain injury, pain, cocaine overdose and convulsions. PMID- 16719808 TI - NMDA receptors are not alone: dynamic regulation of NMDA receptor structure and function by neuregulins and transient cholesterol-rich membrane domains leads to disease-specific nuances of glutamate-signalling. AB - Glutamate receptors of the N-methyl-D-asparate (NMDA-) subtype are tetrameric allosteric and ligand-gated calcium channels. They are modulated by a variety of endogenous ligands and ions and play a pivotal role in memory-related signal transduction due to a voltage-dependent block by magnesium, which makes them Hebbian coincidence detectors. On the structural level NMDA receptors have an enormous flexibility due to seven genes (NR1, NR2A-D and NR3A-B), alternative splicing, RNA-editing and extensive posttranslational modifications, like phosphorylation and glycosylation. NMDA receptors are thought to be responsible for excitotoxicity and subsequent downstream events like neuroinflammation and apoptosis and thus have been implicated in many important human pathologies, ranging from amyotrophic lateral sclerosis, Alzheimer's and Parkinson' disease, depression, epilepsy, trauma and stroke to schizophrenia. This fundamental significance of NMDA receptor-related excitotoxicity is discussed in the context of the developing clinical success of Memantine, but moreover set into relation to various proteomic and genetic markers of said diseases. The very complex localisational and functional regulation of NMDA receptors appears to be dependent on neuregulins and receptor tyrosine kinases in cholesterol-rich membrane domains (lipid rafts), calcium-related mitochondrial feedback-loops and subsynaptic structural elements like PSD-95 (post-synaptic density protein of 95 kD). The flexibility and multitude of interaction partners and possibilities of these highly dynamic molecular systems are discussed in terms of drug development strategies, in particular comparing high affinity and sub-type specific ligands to currently successful or promising therapies. PMID- 16719809 TI - NR2B selective NMDA antagonists: the evolution of the ifenprodil-type pharmacophore. AB - The quest for NR2B subtype selective NMDA antagonists started almost twenty years ago. The structure of ifenprodil, the prototype of this group of compounds, inspired many medicinal chemists in several research units. Different approaches led to the identification of the pharmacophore and several distinct classes of compounds containing this pharmacophore. From these studies a few drug candidates emerged and clinical trials proved the viability of the concept. This article attempted to follow the evolution from ifenprodil, a multiple ligand, to selective NR2B antagonists. PMID- 16719810 TI - Recent advances in the development of NR2B subtype-selective NMDA receptor antagonists. AB - Over activation of the NMDA receptor complex has been implicated in a number of neurological conditions. The use of NMDA antagonists as therapeutic agents has been limited by serious cognitive and motor side effects. Significant efforts have been reported in the development of NR2B subtype-selective antagonists, which have shown efficacy without the side effects observed with nonspecific NMDA antagonists. Classical ifenprodil-like molecules containing benzyl- and phenylpiperidines attached to a phenol or an appropriate isostere by a linker have provided valuable chemical leads as potential therapeutic agents. In this review, recent efforts in the discovery and development of structurally unique NR2B subtype-selective NMDA antagonists that do not fit the classical "ifenprodil like" pharmacophore will be discussed. PMID- 16719811 TI - Targeting PDZ domain proteins for treating NMDA receptor-mediated excitotoxicity. AB - N-methyl-D-aspartate (NMDA) receptors play essential roles in the normal physiology of neurons, and these receptors are also largely responsible for glutamate-induced excitotoxicity. Since treatments of glutamate-induced excitotoxicity by NMDA receptor inhibitors often result in adverse side effects, alternative treatment approaches have been actively sought in recent years. One potential approach is to target proteins and enzymes down stream of the NMDA receptor signaling pathways. Extensive studies in recent years have demonstrated that PDZ domains of PSD-95 play critical roles in scaffolding the NMDA receptor/neuronal nitric oxide synthase pathway. Therefore, PSD-95 PDZ domains become attractive targets for treatment of glutamate-induced overproduction of nitric oxide. The strategy is to develop small compounds that can effectively block protein-protein interactions mediated by the PDZ domains of PSD-95. Biochemical and structural studies of PDZ/target interactions have indicated that developing small molecules to compete with PDZ targets is a feasible approach. We provide an example demonstrating the discovery and further development of small molecules capable of disrupting PSD-95/NMDA receptor and/or PSD-95/neuronal nitric oxide synthase complexes. PMID- 16719812 TI - Relationships between the structure of dexoxadrol and etoxadrol analogues and their NMDA receptor affinity. AB - In the mid 1960s the (dioxolan-4-yl)piperidine derivatives dexoxadrol ((S,S)-1a) and etoxadrol ((S,S,S)-2a) were synthesized. Their pharmacological potential as analgesics, anesthetics and local anesthetics was evaluated in animal models and later on in clinical trials with patients. However, severe side effects including psychotomimetic effects, unpleasant dreams and aberrations stopped the clinical evaluation of dexoxadrol and etoxadrol. Both dioxolane derivatives represent NMDA receptor antagonists, which possess high affinity to the phencyclidine binding site within the NMDA receptor associated ion channel. In this review relationships between the structure of acetalic dexoxadrol analogues and homologues and their affinity toward the phencyclidine binding site of the NMDA receptor are summarized. In particular, high affinity is attained with compounds bearing two phenyl residues or one phenyl residue and an alkyl residue with two or three carbon atoms at the acetalic center. At least one oxygen atom of the oxygen heterocycle is necessary. Instead of the entire piperidine ring aminoalkyl substructures are sufficient for strong receptor interactions. Compounds with a primary amino moiety generally display the highest receptor affinity, whereas tertiary amines possess low affinity. Enlargement of the 1,3-dioxolane ring to a 1,3-dioxane ring or elongation of the oxygen heterocycle / amino group distance results in compounds with considerable NMDA receptor affinity. PMID- 16719813 TI - Tricyclic quinoxalinediones, aza-kynurenic acids, and indole-2-carboxylic acids as in vivo active NMDA-glycine antagonists. AB - This review article describes the development of in vivo active antagonists for the glycine binding site of the N-Methyl-D-Aspartate (NMDA) receptor. There were several difficulties in identifying a class of antagonists with in vivo efficacy and only a few compounds succeeded in emerging with activity in vivo. A series of tricyclic quinoxalinediones was highly potent glycine antagonists in vitro and the derivatives having a zwitterionic moiety including SM-18400 indeed showed in vivo activity. Similarly, tricyclic indole-2-carboxylic acids having a zwitterionic moiety such as SM-31900 were also active in vivo. In fact, SM-18400 and SM-31900 exhibited efficacy in several animal stroke models using intravenous infusion protocols. The practical syntheses of SM-18400 and SM-31900 as well as the novel synthesis of moderately active glycine antagonists, tricyclic azakynurenic acids, were also developed. PMID- 16719815 TI - N-methyl-D-aspartate receptor (NMDA) antagonists as potential pain therapeutics. AB - NMDA receptors are known to be involved in nociceptive transmission and pain processing. Many structurally diverse NMDA antagonists have been reported to have activity in both animal models and clinical models of neuropathic pain. Untoward side effects such as ataxia and sedation have severely limited the clinical uses of this class of potential therapeutics. However, antagonists at the glycine site, NR2B sites and weak-binding channel blockers have demonstrated an improved side effect profile in animal models of pain. These types of compounds may hold potential promise for future pain therapies. This review covers reported pain data surrounding representative examples of NMDA antagonists and provides a current assessment of potential clinical utility. PMID- 16719816 TI - Progress towards validating the NMDA receptor hypofunction hypothesis of schizophrenia. AB - This article describes recent progress towards validation of the N-methyl-D aspartate (NMDA) receptor hypofunction hypothesis of schizophrenia in preclinical models. Schizophrenia, a complex disease characterized by positive, negative and cognitive symptoms, affects 1% of the world population and requires lifelong, daily maintenance therapy. For the last several decades, thinking in this field has been dominated by the hypothesis that hyperfunction of dopamine pathways played a key role in schizophrenia. However, the therapeutic agents developed from this hypothesis have a slow onset of action and tend to improve only the positive symptoms of the disease. The NMDA receptor antagonist PCP has been shown to induce the positive, negative and cognitive symptoms of schizophrenia in healthy patients and cause a resurgence of symptoms in stable patients. These observations led to the NMDA receptor hypofunction hypothesis as an alternative theory for the underlying cause of schizophrenia. According to this hypothesis, any agent that can potentiate NMDA receptor currents has the potential to ameliorate the symptoms of schizophrenia. To date, NMDA receptor currents can be modulated by either direct action on modulatory sites on the NMDA receptor (i.e., the glycine co-agonist binding site) or indirectly by activation of G-protein coupled receptors (GPCRs) known to potentiate NMDA receptor function (i.e., mGluR5). This review will discuss the NMDA receptor hypofunction hypothesis, the NMDA receptor as an emerging target for the development of novel antipsychotic agents and progress towards in vivo target validation with GlyT1 inhibitors and mGluR5 positive allosteric modulators. Other potential targets for modulating NMDA receptor currents (polyamine sites, muscarinic receptors, etc...) will also be addressed briefly. PMID- 16719817 TI - Survival signaling pathways activated by NMDA receptors. AB - N-methyl-D-aspartate receptors (NMDAR) have a recognized role in neuronal plasticity while their excessive activation results in excitotoxic death. Therefore, NMDAR antagonists are considered for neuroprotective interventions. However, there is also an emerging role of NMDAR in supporting neuronal survival. Thus, during CNS development, basal NMDAR activity suppresses neuronal apoptosis while moderate NMDAR activation may, at least under some conditions, protect against excitotoxic/ischemic insults. These suggest that while protecting from excitotoxicity, NMDAR antagonists would also reduce pro-survival activity of NMDAR. Hence, the identification of the switches controlling pro-survival vs. pro excitotoxic outcome of NMDAR stimulation may lead to development of NMDAR antagonists that selectively block the excitotoxicity while enhancing the protective NMDAR signaling. On the other hand, the existence of anti apoptotic/pro-proliferative NMDAR signaling in transformed cells may result in new strategies to attack cancer. This review focuses on the emerging field of neuroprotective signaling mediators that are implicated in pro-survival activity of NMDAR. We discuss the evidence implicating either NR2B or nonNR2B NMDAR in mediating the protection. We also present the reports linking NMDAR-mediated protection to the activation of survival signaling kinases including ERK and Akt, or suppression of a pro-apoptotic kinase, GSK-3beta. The protective role of transcription factors is also discussed. Finally, we review the existing evidence suggesting that NMDAR support survival by regulating the pro-survival trophic factor signaling and/or the cell death machinery. Although NMDAR provide a major survival input to CNS neurons, the NMDAR-activated protective signaling is poorly understood and, therefore, deserves further research effort. PMID- 16719818 TI - The NMDA/D1 receptor complex as a new target in drug development. AB - Dopamine and glutamate have been shown to extensively interact in the striatum, nucleus accumbens, hippocampus and prefrontal cortex, to regulate different physiological functions, including locomotor activity, positive reinforcement, attention and working memory. Although dysfunctions of dopamine transmission have long been identified as critical determinants of neurological and neuropsychiatric disorders, such as Parkinson's disease and schizophrenia, there is now increasing evidence that concurrent alterations of dopamine and glutamate function may play a central role in the pathophysiology of these diseases. Thus, defining the characteristics of dopamine-glutamate interactions may be crucial to identify alternative molecular targets for the development of novel pharmacological tools. At the postsynaptic level, interactions between the dopamine D1 and the glutamate NMDA receptors appear to be particularly relevant. Different mechanisms are involved in this interactions: 1) D1R-dependent, second messenger-mediated phosphorylation of NMDAR subunits; 2) coordinated regulation of receptor trafficking at synaptic sites; 3) formation of an heteromeric D1/NMDA receptor complex. In this paper we review the molecular mechanisms, functional implications and pharmacological significance of D1R/NMDAR interaction via direct protein-protein oligomerization. PMID- 16719819 TI - Competitive Gly/NMDA receptor antagonists. AB - Glutamic acid (Glu) is the major excitatory neurotransmitter in the mammalian central nervous system (CNS) where it is involved in the physiological regulation of different processes. It has been well established that excessive endogenous Glu is associated with many acute and chronic neurodegenerative disorders such as cerebral ischemia, epilepsy, amiotrophic lateral sclerosis (ALS), Parkinson's and Alzheimer's diseases. In addition to the classical competitive glutamate receptor (GluR) antagonists, much effort has been directed toward the development of many different non-competitive antagonists of these receptors and, among them, compounds blocking the glycine site on the NMDA receptor complex (Gly/NMDA) have been widely investigated. Many Gly/NMDA receptor antagonists showed to be potential therapeutic agents in many neurological diseases such as stroke, epilepsy and neuropathic pain. Some of them, endowed also with favourable physicochemical properties and low secondary undesiderable effects, reached clinical trials. PMID- 16719820 TI - Can selective ligands for glutamate binding proteins be rationally designed? AB - A major neurotransmitter, L-Glutamate must be stored, transported and received, and these processes are mediated by proteins that bind this simple yet essential amino acid. Detailed evidence continues to emerge on the structure of Glu binding proteins, which includes both receptors and transporters. It appears that receptors and transporters bind to Glu in different conformations, which may present a pharmacological opportunity. This review will compare and contrast information available on Glu Receptors (AMPA, NMDA, KA and mGlu), excitatory amino acid transporters (EAATs), the system Xc- transporter (XCT) and the vesicular Glutamate transporter (GVT). The cross-reactivity of ligands which have been previously used to characterize the glutamate binding proteins with system Xc- raises some fundamental interpretational issues regarding the mechanisms through which these analogues produce CNS damage. Although at one time it was thought that unraveling selectivity among glutamate binding proteins was an intractable problem, recently the NMDA antagonist (memantine) has been approved for general medical practice for treatment of Alzheimer's disease. Two other agents are in advanced clinical trials: an Ampakine for potential improvement of cognitive disorders, and a selective mGlu agonist for treatment of anxiety. The prospects for unraveling cross-reactivity will be weighed in light of a critical comparison of the glutamate binding protein targets. PMID- 16719821 TI - ACE inhibitor-angiotensin receptor blocker combinations: a clinician's perspective. AB - ACE Inhibitors (ACEI) and angiotensin receptor blockers (ARB) inhibit the renin angiotensin system, but ACEI may do so incompletely when administered as monotherapy at conventional doses. In theory, combining an ACEI and ARB might be beneficial, whereas clinical evidence for this approach in hypertension is lacking. An ACEI-ARB combination is likely to be useful in proteinuric renal disease, but recent experimental evidence suggests that very high dose monotherapy with an ARB may be the best approach. However, the results of large outcome studies for combinations vs. ACEI or ARB monotherapy are still awaited. PMID- 16719822 TI - Topical imiquimod: mechanism of action and clinical applications. AB - Imiquimod is a synthetic imidazoquinoline heterocyclic amine of 240.3 Da (C14H16N4). Imiquimod is a cytokine inducer and a modifier of the innate immune response, as well as acquired antiviral and antitumor immune responses. Imiquimod 5% cream has proven to be an effective treatment for external genital warts, superficial basal cell carcinoma, and actinic keratosis. PMID- 16719824 TI - Regulatory T cells in cancer biology: a possible new target for biochemical therapies. AB - CD4+ CD25+ T cells are essential for maintenance of self-tolerance and therefore have been referred to as regulatory T cells (Treg). Experimental tumor models revealed that Treg are potent inhibitors of an anti-tumor immune response. Treg are expanded in human cancer. Currently, a variety of strategies for the induction of a specific anti-tumor immune response are tested in preclinical and clinical settings. Biochemical strategies modifying and/or depleting Treg in cancer patients for an enhancement of vaccine-based therapeutic concepts will be discussed in detail in this review. PMID- 16719823 TI - Antioxidative actions of statins: potential mechanisms for antiathersclerotic effects. AB - Inhibitors of 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase (statins) are widely used for the prevention of atherosclerotic diseases. The effects of statins on the generation of reactive oxygen species (ROS) by in vitro and in vivo were studied. Administration of statins significantly decreased ROS generation in vitro and in vivo. PMID- 16719825 TI - Rational design of non-hydroxamate histone deacetylase inhibitors. AB - While most inhibitors of histone deacetylases (HDACs) are hydroxamic acid derivatives, several non-hydroxamates have recently been developed as inhibitors and attracted quite a deal of attention. In this review, we present the rational design, inhibitory effect and antiproliferative activity of non-hydroxamate HDAC inhibitors. PMID- 16719826 TI - Ether lipids as anticancer agents: focus on non-phosphorus cationic glycerolipids. AB - This review focuses on the synthesis and mechanisms of antitumor activity of cationic non-phosphorus analogs of edelfosine. The role of variable length and mode of conjugation of the spacer group, the types of cationic 'head', and the length of the substituent at C2 atom of the glycerol backbone are discussed, providing the basis for rational design of lipophilic anticancer drugs, in particular, for elimination of multidrug resistant cells. PMID- 16719827 TI - Small molecule and biologic modulators of the immune response to hepatitis C virus. AB - Hepatitis C virus represents a major global health problem, with approximately 3% of the world population infected. Immune-response modifiers represent the standard of care, given the lack of approved antiviral agents having direct activity against the viral proteins. Although in recent years, improvements in therapy have been attained by combined treatment with pegylated interferon and ribavirin, the discovery and development of next-generation small molecule and biologic agents is ongoing. Several of these newer therapeutics are focused on modulating Toll-like receptors, interferon-alpha signaling, and the pro inflammatory cytokine balance. A comprehensive account of the lead compounds in development, the bioassays used for optimization of these immune response modifiers and their clinical status is presented. PMID- 16719828 TI - Reconsideration of Japanese traditional herbal medicine: new field of research and clinical medicine. AB - Japanese traditional herbal medicine, which is known as "Kampo", has received increasing attention as an alternative approach to the treatment of various diseases. In order to safely and effectively apply Kampo, it is essential to understand oriental diagnostics. Herein, an outline of such diagnostics is introduced, and current trends in Kampo research are reviewed. PMID- 16719829 TI - Molecular insights into the mechanisms of HIV-1 reverse transcriptase resistance to nucleoside analogs. AB - The causative agent of acquired immunodeficiency syndrome, HIV-1, depends on one of its enzymes, reverse transcriptase, to copy its single stranded RNA genome into a double stranded DNA nucleic acid suitable for integration in the host cell genome. In the last two decades, the advances in the knowledge of the kinetic mechanism of reverse transcription and in the determination of the crystallographic structures for the complexes of the enzyme with substrates and products were huge. However, all of this knowledge resulted in the design of RT inhibitors for which the virus, after a short period of exposure, becomes less susceptible, due to the development of resistance. The development of resistance is caused by the high frequency of viral mutation and the toxicity of those same drugs. Therefore, a closer look at all the available information might shed some light into this subject and help to develop new strategies to overcome the lack of long term clinical efficiency of these drugs. Here, we present a critical atomic level study of all the mutations that have been detected and reported so far, as a reaction of the enzyme to counteract the action of the inhibitors. PMID- 16719830 TI - Novel anti-viral therapy: drugs that block HIV entry at different target sites. AB - Drugs that block the entry of human immunodeficiency virus type 1 (HIV-1) into host cells abrogate the establishment of a productive infection and should ideally diminish the chances of HIV-1 developing resistance. This review will give an overview of the mechanism by which the envelope glycoprotein mediates HIV 1 entry and will summarize current drug developments. PMID- 16719831 TI - Fine tuning of PPAR ligands for type 2 diabetes and metabolic syndrome. AB - Type 2 diabetes mellitus (T2DM) is highly prevalent chronic disease. Recently, many biological targets are discovered for treatment of this disease. The identification of the nuclear hormone receptor peroxisome proliferator activated receptors (PPAR) and their subtypes alpha, gamma and delta or beta as targets for controlling lipid, glucose and energy homeostasis has proved to be exciting. As hyperlipidaemia, obesity and insulin resistance are independent risk factors for coronary heart disease and macrovascular complications of diabetes; new agents that increase insulin sensitivity as well as decrease hyperlipidaemia by distinct yet complementary mechanism are being studied as they may provide improved therapy for T2DM and related disorders. In this article, we review highly potent PPARgamma agonists, PPARalpha/gamma dual agonists, PPAR pan agonists, alternative PPAR ligands like partial agonists or selective PPAR modulators (SPPARMs) and antagonists from a chemist point of view. PMID- 16719832 TI - Pharmacological and clinical studies on purine nucleoside analogs--new anticancer agents. AB - More recently, three novel purine nucleoside analogs, including clofarabine, nelarabine and immucillin H, have been introduced into clinical trials. These agents have different metabolic properties, novel mechanism of action, and are undergoing phase I-II clinical studies for the treatment of hematopoietic malignancies. Pharmacology and anticancer activity of PNA are the subjects of this review. PMID- 16719833 TI - The emergence of plasma membrane calcium pump as a novel therapeutic target for heart disease. AB - The plasma membrane calcium/calmodulin dependent ATPase (PMCA) is a calcium extruding enzymatic pump important in the control of intracellular calcium concentration. PMCA is the only system for calcium extrusion in the majority of cells. In excitable cells such as cardiomyocytes however, PMCA has been shown to play only a minor role in calcium homeostasis. In these cells the main mechanism of calcium extrusion is the sodium calcium exchanger. However, increasing evidence points to an important role for PMCA in signal transduction; in particular in the nitric oxide signalling pathway. In this review we will discuss recent advances that support a key role for PMCA in signal transduction and the potential for therapeutic targeting of this molecule in the treatment of cardiac diseases. PMID- 16719834 TI - Recent advances in clarifying prion protein functions using knockout mice and derived cell lines. AB - Considerable information on the functions of prion protein (PrP) has been accumulated. One experimental approach is the use of PrP gene-knockout mice and derived cell lines. This approach has contributed to elucidating the functions of cellular prion protein (PrP(C)), such as its anti-oxidative and anti-apoptotic roles. This review will introduce the recent advances in prion biology made possible by the availability of these tools. PMID- 16719835 TI - The therapeutic potential of Angiotensin-(1-7) as a novel Renin-Angiotensin System mediator. AB - In this review, we show Angiotensin-(1-7) as a novel Renin Angiotensin System mediator that antagonizes cardiovascular and proliferative effects of Angiotensin II and exerts complex renal actions. We also speculate the possibility of new drugs for the treatment of cardiovascular, genitourinary and hepatic diseases by interfering with ACE2-Angiotensin-(1-7)-Mas axis. PMID- 16719836 TI - Differential analysis of secondary metabolites by LC-MS following strain improvement of Streptomyces lydicus AS 4.2501. AB - Metabolite variations in a high-yielding mutant and its parent strain were studied by comparative LC-MS analysis after strain improvement. Streptomyces lydicus AS 4.2501-P28, a propionate-resistant mutant isolated by the high frequency screening method using the principle of eliminating precursor inhibition effects, showed an increase of 267% in streptolydigin titre over the starting strain. Culture extracts of this mutant and its parent strain were analysed in parallel by an LC-MS technique, including full scan and extracted-ion scan, ESI-MS (electrospray-ionization MS) detection, DAD (diode-array detection) and MS2 (tandem MS) measurement. The main metabolic variations were obviously found in intermediates, metabolites and biosynthetic pathways: two unknown metabolites with the molecular [M-H]- ions at m/z 423.3 and 687.2, corresponding to two branch pathways, were blocked in the mutant, and the accumulation of a significant intermediate at m/z 363.1 [M-H]- decreased dramatically in the mutant cultures, resulting in the overproduction of streptolydigin (an antibiotic that inhibits prokaryotic RNA polymerase) in the mutant. Ion fragmentations of the tandem-MS spectra provided experimental evidence for the structural characterization of the three compounds obtained. In comparison with the traditional methods, comparative LC-MS analysis was rapid, sensitive and suitable for characterizing intermediates, metabolites and pathways for elucidation of the metabolic alterations after the isolation of improved strains. PMID- 16719837 TI - Immobilized protein ZZ, an affinity tool for immunoglobulin isolation and immunological experimentation. AB - At present, the common tool for affinity purification of IgG is immobilized Protein A, which is separated from native cell-wall components of Staphylococcus aureus. It is complicated and costly to prepare natural Protein A. ZZ protein is a synthetic Fc region-binding domain originated from B domain of Protein A. In the present study, recombinant ZZ protein with a hexahistidine tag at the N terminus was expressed in BL21 (DE3) under the control of T7 promoter. The protein was purified through one-step Ni2+ chelating affinity chromatography at a yield of 50 mg of protein/litre of culture. Then it was covalently coupled with Sepharose 4B with butane-1,4-diol diglycidyl ether. The protein ZZ-Sepharose 4B resin exhibited good performance in affinity purification of IgG, as well as in capturing the protein-interacting complexes in immunoprecipitation experiments. Compared with natural Protein A, the expression and purification of ZZ protein at high yield are very simple and low-cost. At this point, extensive applications of protein ZZ in immunoassays are practicable and to be anticipated. PMID- 16719838 TI - Biosynthesis of hyaluronan: direction of chain elongation. AB - The mechanism of hyaluronan biosynthesis in vertebrates had been proposed to occur at the reducing end of growing chains. This mechanism was questioned because a recombinant synthase appeared to add new monosaccharides to the non reducing end. I reinvestigated this problem with membranes from the eukaryotic B6 cell line. The membranes were incubated with UDP-[3H]GlcNAc and UDP-[14C]GlcA to yield differentially labelled reducing terminal and non-reducing terminal domains. Digestion of the product with a mixture of the exoglycosidases beta glucuronidase and beta-N-acetylglucosaminidase truncated the hyaluronan chain strictly from the non-reducing end. The change in 3H/14C ratio of the remaining hyaluronan fraction, during the course of exoglycosidase digestion, confirmed the original results that the native eukaryotic synthase extended hyaluronan at the reducing end. This mechanism demands that the UDP-hyaluronan terminus is bound to the active site within the synthase and should compete with the substrates for binding. Accordingly, increasing substrate concentrations enhanced hyaluronan release from the synthase. A model is proposed that explains the direction of chain elongation at the reducing end by the native synthase and at the non reducing end by the recombinant synthase based on a loss of binding affinity of the synthase towards the growing UDP-hyaluronan chain. PMID- 16719839 TI - In vivo contribution of amino acid sulfur to cartilage proteoglycan sulfation. AB - Cytoplasmic sulfate for sulfation reactions may be derived either from extracellular fluids or from catabolism of sulfur-containing amino acids and other thiols. In vitro studies have pointed out the potential relevance of sulfur containing amino acids as sources for sulfation when extracellular sulfate concentration is low or when its transport is impaired such as in DTDST [DTD (diastrophic dysplasia) sulfate transporter] chondrodysplasias. In the present study, we have considered the contribution of cysteine and cysteine derivatives to in vivo macromolecular sulfation of cartilage by using the mouse model of DTD we have recently generated [Forlino, Piazza, Tiveron, Della Torre, Tatangelo, Bonafe, Gualeni, Romano, Pecora, Superti-Furga et al. (2005) Hum. Mol. Genet. 14, 859-871]. By intraperitoneal injection of [35S]cysteine in wild-type and mutant mice and determination of the specific activity of the chondroitin 4-sulfated disaccharide in cartilage, we demonstrated that the pathway by which sulfate is recruited from the intracellular oxidation of thiols is active in vivo. To check whether cysteine derivatives play a role, sulfation of cartilage proteoglycans was measured after treatment for 1 week of newborn mutant and wild-type mice with hypodermic NAC (N-acetyl-L-cysteine). The relative amount of sulfated disaccharides increased in mutant mice treated with NAC compared with the placebo group, indicating an increase in proteoglycan sulfation due to NAC catabolism, although pharmacokinetic studies demonstrated that the drug was rapidly removed from the bloodstream. In conclusion, cysteine contribution to cartilage proteoglycan sulfation in vivo is minimal under physiological conditions even if extracellular sulfate availability is low; however, the contribution of thiols to sulfation becomes significant by increasing their plasma concentration. PMID- 16719840 TI - Isolation of neural crest-derived stem cells from rat embryonic mandibular processes. AB - BACKGROUND INFORMATION: Substantial evidence indicates the existence of NCSCs (neural crest-derived stem cells) in embryonic mandibular processes; however, they have not been fully investigated or isolated. The aim of the present study was to isolate stem cells from mandibular process during embryonic development by MACS (magnetic-activated cell sorting). The findings show that the cells are multipotent and self-renewing. RESULTS: LNGFR (low-affinity nerve-growth-factor receptor)+ cells were isolated from rat embryonic mandibular processes by MACS. The cells were grown in clonal culture by limiting dilution to assess their developmental potential. Clone analysis indicated that, first, LNGFR+ cells are multipotent, being able to generate at least neurons and Schwann cells, similar to peripheral neural crest stem cells. Secondly, multipotent LNGFR+ cells generate multipotent progenies, indicating that they are capable of self-renewal and therefore are stem cells. Thirdly, manipulation of the medium supplementation alters the fate of the isolated LNGFR+ cells. CONCLUSIONS: These results indicate that LNGFR antibodies label NCSCs with high specificity and purity, and suggest that positive selection using these antibodies may become the method of choice for obtaining multipotent cells from rat embryonic mandibular processes for tissue engineering or regenerative therapeutic use. PMID- 16719841 TI - Early alterations in myocardia and vessels of the diabetic rat heart: an FTIR microspectroscopic study. AB - Diabetes mellitus is associated with a high incidence and poor prognosis of cardiovascular disease. The aim of the present study was to examine the effect of relatively short-term (5 weeks) Type I diabetes on the left ventricle, the right ventricle and the vessel (vein) on the left ventricle of the myocardium at molecular level by FTIR (Fourier-transform infrared) microspectroscopy. The rats were categorized into two groups: control group (for the left ventricle myocardium, n=8; for the right ventricle myocardium, n=9; for the vein, n=9) and streptozotocin-induced diabetic group (for the left ventricle myocardium, n=7; for the right ventricle myocardium, n=9; for the vein, n=8). Two adjacent cross sections of 9 microm thickness were taken from the ventricles of the hearts in two groups of rats by using a cryotome. The first sections were used for FTIR microspectroscopy measurements. The second serial sections were stained by haematoxylin/eosin for comparative purposes. Diabetes caused an increase in the content of lipids, an alteration in protein profile with a decrease in alpha helix and an increase in beta-sheet structure as well as an increase in glycogen and glycolipid contents in both ventricles and the vein. Additionally, the collagen content was found to be increased in the vein of the diabetic group. The present study demonstrated that diabetes-induced alterations in the rat heart can be detected by correlating the IR spectral changes with biochemical profiles in detail. The present study for the first time demonstrated the diabetes-induced alterations at molecular level in both ventricle myocardia and the veins in relatively short-term diabetes. PMID- 16719843 TI - Metabolic syndrome: fact or fiction. PMID- 16719842 TI - Endogenous ADP-ribose enables calcium-regulated cation currents through TRPM2 channels in neutrophil granulocytes. AB - TRPM2 (transient receptor potential melastatin 2) is a Ca2+-permeable cation channel gated by ADPR (ADP-ribose) from the cytosolic side. To test whether endogenous concentrations of intracellular ADPR are sufficient for TRPM2 gating in neutrophil granulocytes, we devised an HPLC method to determine ADPR contents in HClO4 cell extracts. The reversed-phase ion-pair HPLC method with an Mg2+ containing isocratic eluent allows baseline resolution of one ADPR peak. Intracellular ADPR concentrations were approx. 5 muM in granulocytes and not significantly altered by stimulation with the chemoattractant peptide fMLP (N formylmethionyl-leucylphenylalanine). We furthermore determined intracellular concentrations of cADPR (cyclic ADPR) with a cyclase assay involving enzymatic conversion of cADPR into NAD+ and fluorimetric determination of NAD+. Intracellular cADPR concentrations were approx. 0.2 microM and not altered by fMLP. In patch-clamp experiments, ADPR (0.1-100 microM) was dialysed into granulocytes to analyse its effects on whole-cell currents characteristic for TRPM2, in the presence of a low (<10 nM) or a high (1 microM) intracellular Ca2+ concentration. TRPM2 currents were significantly larger at high than at low [Ca2+] (e.g. -225+/-27.1 versus -7+/-2.0 pA/pF at 5 muM ADPR), but no currents at all were observed in the absence of ADPR (ADPR concentration < or =0.3 microM). cADPR (0.1, 0.3 and 10 microM) was without effect even in the presence of subthreshold ADPR (0.1 microM). We conclude that ADPR enables an effective regulation of TRPM2 by cytosolic Ca2+. Thus ADPR and Ca2+ in concert behave as a messenger system for agonist-induced influx of Ca2+ through TRPM2 in granulocytes. PMID- 16719844 TI - Pediatric systemic lupus erythematosus: management issues in primary practice. AB - PURPOSE: To provide nurse practitioners (NPs) with an update on pediatric systemic lupus erythematosus (SLE) with an emphasis on management considerations for primary care practitioners. DATA SOURCES: An extensive literature review was conducted using both Medline and CINAHL databases. Research articles reflecting the most compelling findings were included in this review. CONCLUSIONS: NPs who care for children with SLE may be able to prevent or delay the morbidities associated with this disease and its treatments, if they keep abreast of the new information evolving in this realm of rheumatologic diseases. IMPLICATIONS FOR PRACTICE: As more is learned about pediatric SLE, better treatments have evolved such that the survival rates have increased. The primary care of pediatric SLE patients that is focused on preventing or delaying SLE morbidities may help to restore, maintain, or improve the quality of life for these patients. PMID- 16719845 TI - Modifiable risk factors for impaired fertility in women: what nurse practitioners need to know. AB - PURPOSE: To provide an overview of impaired fertility in childbearing-aged women, to review the current research on modifiable lifestyle risk factors implicated in its development, and to suggest strategies for nurse practitioners (NPs) to assist women in behavioral changes that will allow them to protect their fertility. DATA SOURCES: Original research articles and comprehensive review articles identified through Medline, CINAHL, and OVID databases. CONCLUSIONS: Research has shown that advancing age, a history of a sexually transmitted infection and/or pelvic inflammatory disease, extremes of body weight, and tobacco and caffeine use are potentially modifiable risk factors in the development of impaired fertility. IMPLICATIONS FOR PRACTICE: NPs must be aware of the link between these behaviors and the development of impaired fertility in order to assist women in preserving their fertility. Individual counseling, education, and community-wide education strategies are discussed. PMID- 16719846 TI - Pilot study: nurse practitioner communication and the use of recommended clinical preventive services. AB - PURPOSE: The purpose of the pilot study was to determine the feasibility of investigating nurse practitioners' (NPs) use of clinical preventive services (CPS) in practice and the influence of provider-patient communication in achieving the CPS outcome. NP-patient communication styles were also explored. DATA SOURCES: Five NPs were audio-taped each during one patient encounter. Using manifest content analysis, the transcripts were evaluated for the presence of CPS and communication styles. CONCLUSION: Because this was a pilot study and the sample size was small, no generalizations could be made. The presence of health promotion and disease prevention activities, as evaluated by the use of recommended CPS, was present in only one NP-patient encounter. NPs did use patient-centered communication style in all the encounters. No correlations could be made to link communication styles with CPS. IMPLICATIONS FOR PRACTICE: Further study with a larger sample size needs to be done. NPs may not be using health promotion and disease prevention, as demonstrated by CPS, in actual practice. This may be an area that needs greater emphasis in education and practice. PMID- 16719847 TI - When I go in to wake them ... I wonder: parental perceptions about congenital long QT syndrome. AB - PURPOSE: Congenital long QT syndrome (LQTS) affects an estimated 1 in 5,000 persons, is characterized by QT interval prolongation, and has a clinical presentation ranging from asymptomatic longevity to sudden death in the young as the initial event. The purpose of this study was to describe the experiences of parents who have a child or children with LQTS. Information from parents of children with this diagnosis can provide insight to healthcare providers who care for these families. DATA SOURCES: The literature reveals that very little is known about the psychosocial aspects of this potential sudden death syndrome. This was a secondary analysis of a 2002 qualitative phenomenological primary study done to explore fear of death and quality of life for 58 patients with LQTS. The secondary study analyzed responses derived from 31 parents of children with LQTS. CONCLUSIONS: We speculated this far-reaching clinical spectrum would arouse fear and uncertainty for a parent of a child with LQTS. Results of the study revealed that parents with young children described fear of their children dying and strategies they used to manage their fear, as well as frustrations about lack of knowledge of LQTS among healthcare providers. When the diagnosis of LQTS is established during adolescence, the impact on the lives of children and their families is more significant. IMPLICATIONS FOR PRACTICE: In order to support families experiencing the stress of living daily with someone with LQTS, healthcare providers including nurse practitioners need a better understanding of the symptoms, diagnosis, management, and lifestyle implications of LQTS. Further studies are needed to understand the long-term psychosocial effects of children on beta-blockers, children with implantable cardioverter defibrillators, and children, adolescents, and young adults who survive a sudden death event. PMID- 16719848 TI - Prescribing trends by nurse practitioners and physician assistants in the United States. AB - PURPOSE: As an important step in analyzing the role of nurse practitioners (NPs) and physician assistants (PAs), we examined their prescribing behavior. The intent is to study the characteristics of providers and patients, and the type of prescriptions written by NPs and PAs in primary care and to compare these activities to physicians. DATA SOURCES: The National Ambulatory Medical Care Survey (NAMCS) database was examined for prescriptions written by primary care clinicians (family and general medicine, internal medicine, and general pediatrics). A representative sample of 88,346 primary care visits over a 6-year period (1997-2002) was analyzed in which a prescription was written by an NP, a PA, or a physician in an urban or rural setting. CONCLUSIONS: The characteristics of all the patients seen were similar for geographical region of visit, age, and gender, but differed by ethnicity and race. An NP or a PA was the provider of record for 5% of the primary care visits in the NAMCS database. The three clinician types were likely to write at least one prescription for 70% of all visits, and the mean number of prescriptions was 1.3-1.5 per visit (range 0-5) depending on the provider. PAs were more likely to prescribe a controlled substance for a visit than a physician or an NP (19.5%, 12.4%, 10.9%, respectively). Only in nonmetropolitan settings did differences emerge. In rural areas, NPs wrote significantly more prescriptions than physicians and PAs. IMPLICATIONS FOR PRACTICE: We suggest that NPs and PAs may provide a role that is similar to that of physicians in primary care based on prescribing behavior. The prescribing behavior of PAs and NPs parallels that of physicians by the number of medications per visit, the types of therapeutic classes, and the type of patient. However, in nonmetropolitan areas, prescribing differences emerge between the three types of providers that bear further exploration. PMID- 16719849 TI - Community-acquired methicillin-resistant Staphylococcus aureus skin infections: report of a local outbreak and implications for emergency department care. AB - PURPOSE: To report a local outbreak of community-associated methicillin-resistant Staphylococcus aureas (CA-MRSA), discuss the incidence and the high rates of CA MRSA-related skin and soft tissue infections in our outpatient and emergency department (ED) population, and also review the diagnosis and management of CA MRSA in outpatients. DATA SOURCES: Case report, diagnostic evidence, scientific literature, published treatment guidelines, and a retrospective chart review. CONCLUSIONS: Clinical suspicions of high rates of CA-MRSA-related skin and soft tissue infections in our institution's outpatient and ED population was supported. Hospital-acquired MRSA (HA-MRSA) was also more prevalent than expected. Recognizing high-risk settings, patients and clinical features of CA MRSA is essential for early identification and proper management. The most common clinical manifestation of CA-MRSA we identified was a skin or soft tissue infection. IMPLICATIONS FOR PRACTICE: It is important that clinicians have an awareness of high-risk patients, perform routine culturing of soft tissue infections, and prescribe antibiotics based on culture and sensitivities. Awareness, prevention, early diagnosis, and implementation of effective antibiotic management by nurse practitioners can help limit an epidemic of CA MRSA. PMID- 16719851 TI - Cervical screening: striving for outcome beyond expectation. PMID- 16719852 TI - Rapid rescreening of cervical smears as a quality control method in a high-risk population. AB - OBJECTIVE: Cancer of the cervix is one of the commonest cancers in South Africa. Accurate cytological diagnosis is one of the prerequisites for an effective cervical screening programme and requires the implementation of appropriate quality assurance modalities. This study was undertaken to determine if rapid review of reportedly negative cervical smears is a useful internal quality assurance modality in an unscreened population with very high rates of cervical carcinoma. METHOD: Approximately 26% of all cervical smears received at the study institution between 1 January 1998 and 31 December 2003, and initially reported as negative or inadequate, underwent rapid review. RESULTS: A total of 62,866 (26%) cervical smears out of 241,796 reportedly negative or inadequate cervical smears underwent rapid review. An amended report was sent out in 373 (0.59%) of these 62,866 cervical smears. This included 101 cases of high-grade squamous intraepithelial lesion (HSIL) and high-grade atypical squamous cells (ASC-H), 143 low-grade squamous intraepithelial lesions, 54 atypical squamous cells of undetermined significance (ASC-US) and 33 atypical glandular cells that were not reported initially. The false-negative proportion for HSIL and ASC-H (combined) in this study was 5.76%. No squamous cell carcinomas were diagnosed on rapid review but one patient with HSIL/ASC-H on review had squamous cell carcinoma on biopsy. Three cytotechnologists had a lower sensitivity of primary screening and required retraining. CONCLUSIONS: Rapid review is beneficial as an internal quality assurance modality in an unscreened high-risk population and increases the detection of women with significant cervical lesions requiring treatment. The relatively low cost of rapid review compared with other rescreening modalities makes this an attractive option in low resource settings. PMID- 16719853 TI - Evaluation of 100% rapid rescreening of negative cervical smears as a quality assurance measure. AB - OBJECTIVE: The objective of this study was to compare the performance of 100% rapid rescreening, 10% random rescreening and the review of smears selected on the basis of clinical criteria, as a method of internal quality control of cervical smears classified as negative during routine screening. METHODS: A total of 3149 smears were analysed, 173 of which were classified as positive and 2887 as negative, while 89 smears were considered unsatisfactory. The smears classified as negative were submitted to 100% rapid rescreening, 10% random rescreening, and rescreening based on clinical criteria. The rescreening stages were blinded and results were classified according to the Bethesda 2001 terminology. Six cytologists participated in this study, two of whom were responsible for routine screening while the other four alternated in carrying out rescreening so that no individual reviewed the same slide more than once. RESULTS: The 100% rapid rescreening method identified 92 suspect smears, of which 42 were considered positive at final diagnosis. Of the 289 smears submitted to the 10% rescreening method, four were considered abnormal but only one was confirmed positive in the final diagnosis. Of the 690 smears rescreened on the basis of clinical criteria, 10 were considered abnormal and eight received a positive final diagnosis. CONCLUSIONS: The 100% rapid rescreening method is more efficient at detecting false-negative results than 10% random rescreening or rescreening on the basis of clinical criteria, and is recommended as an internal quality control method. PMID- 16719854 TI - Rapid cervicovaginal smear screening: method of quality control and assessing individual cytotechnologist performance. AB - AIM: To validate the method of rapid screening (RS) in the detection of cervical lesions and false-negative results as well as in quality control of cytotechnologist performance. MATERIAL AND METHODS: The RS method was validated on Papanicolaou-stained and initially conventionally analysed vaginal, cervical and endocervical (VCE) smears collected in an opportunistic programme for the detection of cervical carcinoma. The study included 3680 VCE smears from the Department of Gynaecologic Cytology, University Department of Gynaecology and Obstetrics, Zagreb University Hospital Center, Zagreb and from the Department of Clinical Cytology, Osijek University Hospital, Osijek. Histologically verified abnormal findings accounted for 10% of the study samples. Thirteen cytotechnologists, with no previous experience in RS, performed the test. Each slide was examined using the 'step' technique for 1.5 minutes, the findings were classified as negative or abnormal, and the abnormal ones were also classified according to differential cytological diagnosis. The results were compared with those obtained on initial screening. Abnormal findings from a group of initially negative findings were reanalysed using conventional methods to make definitive cytological diagnosis. RESULTS: RS yielded a sensitivity of 83.7%, specificity of 93.7%, positive predictive value of 62.4%, negative predictive value of 97.9% and diagnostic accuracy of 92.6%. Relative to the initial abnormal differential cytological diagnosis, the diagnostic value of RS increased with lesion severity [54.8%, 68.0% and 91.3% for cervical intraepithelial neoplasia (CIN) I, CIN II and CIN III respectively]. RS detected 38 additional positive findings; 94.2% of these were atypical squamous cells of undetermined significance (ASCUS)/abnormal glandular cells undetermined significance (AGUS) and CIN I. The rate of additional positive findings was 1.14% (38/3135). The false-negative rate of initial screening was 9.4% (38/406), and individual cytotechnologist sensitivity was 60.0-100.0%. CONCLUSION: RS could be introduced as an efficient method of quality control to improve the sensitivity of cytological screening as well as for quality control of cytotechnologist performance. PMID- 16719855 TI - Review of cervical smears from 76 women with invasive cervical cancer: cytological findings and medicolegal implications. AB - OBJECTIVE: To review cervical smears from 76 women which were taken prior to the diagnosis of invasive cervical cancer and to determine the appropriateness of the cytology reports issued on the smears. METHODS: Cervical smears, clinical records, cervical smear history and cytology reports from 76 women with invasive cervical cancer were reviewed. After microscopic review of the cervical smears, the cases were divided into two groups: Group 1 comprised 50 women who were found to have had at least one false-negative (F/N) smear report prior to the diagnosis of invasive cervical cancer. Group 2 comprised 26 women for whom no evidence of F/N reporting was found. RESULTS: A total of 209 cervical smears from the 50 women in group 1 were available for review (range 2-12 smears per woman); 100 of the 209 smears were considered to have been reported appropriately. Ninety-seven smears which had been reported originally as negative or inadequate were found, on review, to contain numerous severely dyskaryotic cells and were reclassified as F/N smears. All of the 50 women had at least one F/N smear and 29 had two or more. Twelve smears from eight women contained only a few (<200 severely dyskaryotic cells). Forty women developed invasive squamous carcinoma and 10 developed invasive adenocarcinoma. The stage at diagnosis ranged from 1A to stage 4. Seventy-one smears from the 24 women in group 2 were available for review (range 1-15 smears per woman). In two cases included in group 2, no smears were provided for review as the smears had been lost or mislaid. Review of the 71 smears confirmed the accuracy of the original cytological classification of the smear. Nineteen women were diagnosed with squamous cancer, two microinvasive cancer, one glassy cell, two adenocarcinomas, and one with adenosquamous carcinoma. One women was found to have an embryonal rhabdomyosarcoma of the corpus uteri involving the cervix. DISCUSSION/CONCLUSION: The medicolegal implications are discussed in the light of the above findings. Evidence of breach of duty of care was presented in all 50 cases in group I although causation was not established in every case. There was no evidence of failure of duty of care in terms of the standard of the cervical cytology reports issued or standard of clinical management in 17 of the 26 cases in group 2. However, in seven of the 26 cases in group 2, clinical management of the case was substandard due to failure to investigate symptoms of irregular bleeding regardless of a negative cytology report (two cases), failure to act upon a suspicious smear report or consecutive inadequate smear reports (two cases), failure of follow-up after treatment of CIN3 (two cases) and histological misdiagnosis (one case). PMID- 16719856 TI - Ultrasound guidance improves the adequacy of our preoperative thyroid cytology but not its accuracy. AB - AIMS: Our thyroid cytology audit results of 1990-1995 showed an unsatisfactory rate of 43.1% and prediction of neoplasia with a sensitivity of 86.8%. Increasingly, ultrasound scan (USS)-guided core sampling for cytology is proving a valuable tool instead of freehand fine needle aspiration (FNA) or following unsatisfactory freehand FNA. We present the results of freehand FNA and USS guided core samples for cytology in two separate patient groups in our centre. METHODS: Patients who had a thyroid resection and preoperative thyroid cytology in our institution between 1996 and 2002 were included. The histological diagnoses were correlated with the preceding cytology results. RESULTS: A total of 450 FNAs were performed on 394 patients. Freehand FNAs were performed for 348 (77.3%) samples and USS-guided core for 102 (22.7%) samples; 121 (26.8%) were repeat aspirates performed on 45 patients. Using aspiration cytology (AC) grading, freehand FNA was cytologically inadequate (AC0 or AC1) in 34.8% cases whereas USS-guided core was inadequate in 17.6% cases (P = 0.001). Freehand FNA (AC3, AC4, AC5) predicted neoplasia with a sensitivity of 83.2%, specificity of 46.6%, accuracy of 63.0%, positive predictive value of 56.0% and negative predictive value of 77.1%. USS-guided core sample for cytology (AC3, AC4, AC5) predicted neoplasia with a sensitivity of 93.5%, specificity of 26.0%, accuracy of 51.9%, positive predictive value of 43.9% and negative predictive value of 86.7%. CONCLUSIONS: Although USS-guided core provides more satisfactory specimens than freehand FNA, in our centre it does not provide increased accuracy. PMID- 16719857 TI - Fine needle aspiration cytology of cystic partially differentiated nephroblastoma of the kidney. PMID- 16719858 TI - Pituitary adenoma-neuronal choristoma (PANCH): cytomorphology on smear preparation. PMID- 16719859 TI - Granular cell tumour of the neurohypophysis on cytological squash preparations. PMID- 16719860 TI - The cytology of pulmonary well-differentiated foetal adenocarcinoma. PMID- 16719861 TI - Cytomorphology of secretory hyperplastic (lactational) nodule breast. PMID- 16719862 TI - Leukaemia cutis in T-cell acute lymphoblastic leukaemia. PMID- 16719863 TI - Ultraviolet B radiation suppresses Langerhans cell migration in the dermis by down-regulation of alpha4 integrin. AB - BACKGROUND/PURPOSE: Ultraviolet B (UVB) radiation affects the migration and function of epidermal Langerhans cells (LC) and causes immunosuppression of contact hypersensitivity. It is known that LC leaves the epidermis after exposure to UVB. To know the behavior of LC in the dermis after UVB radiation, we studied the effect of UVB radiation on the expression of integrin families on freshly isolated or cultured murine LC. We also examined whether UVB radiation affects the migration of LC to secondary lymphoid tissue chemokine (SLC/6Ckine). METHODS: Integrin expressions of murine LC cultured in epidermal cell suspension were analyzed using flowcytometry. We used murine LC sorted flowcytometrically for binding assay to extracellular matrix and for migration assay to chemokine. Skin explant assay and immnohistochemical staining for 'cords formation' were performed as previously described. RESULTS: Twenty and 40 mJ/cm2 of UVB radiation down-regulated the expression of alpha4 integrin on 24 h-cultured LC, but not that of alpha6, beta1, or beta4 integrin. The number of cultured LC adhered to fibronectin, a ligand for alpha4 integrin, was decreased after UVB irradiation, while that to laminin, a ligand for alpha6 integrin, was not influenced. UVB radiation reduced the number of migrating LC to SLC. Furthermore, skin sheet explant experiments showed that UVB radiation inhibited the 'cords' formation in dermal vessels of the 48 h-cultured skin. CONCLUSIONS: These data suggest that UVB radiation may suppress the migration of LC from the dermis to lymphatic vessels. UVB radiation may downregulate the adherence of LC to dermal fibronectin and migration to SLC, and consequently suppress the migration of LC from the UVB irradiated dermis to lymphatics. PMID- 16719864 TI - In vivo persistent pigment darkening method: a demonstration of the reproducibility of the UVA protection factors results at several testing laboratories. AB - BACKGROUND/PURPOSE: The aims of the present studies were to check that persistent pigment darkening (PPD) method can produce accurate and reproducible results for high UVA protection factors (UVAPF) and to provide data on the variability between laboratories and on the influence of skin types. METHODS: The Japanese Cosmetic Industry Association (JCIA) PPD method was used to determine the UVAPF in different laboratories of different sunscreen formulations with increasing UVAPF. Two formulations were tested at seven independent laboratories and five products within two laboratories. The influence of skin types on the UVAPF of two products was tested within one laboratory on two panels of volunteers. All laboratories complied with the JCIA method specifications. RESULTS: Reproducible results have been obtained between the different labs and a low and satisfactory variability was achieved with a panel size of 10 subjects. Furthermore, skin type was demonstrated to have no influence within the defined selection criteria. CONCLUSIONS: From this multiple center testing, the PPD method has been shown to be appropriate for testing sunscreen formulations with UVAPFs above 20. It is reasonable to expect that test results will be consistent if an identical protocol is used between laboratories. PMID- 16719865 TI - The inhibition spectrum of solar urticaria suppresses the wheal-flare response following intradermal injection with photo-activated autologous serum but not with compound 48/80. AB - BACKGROUND: The inhibition spectrum (IS) in solar urticaria was identified mainly in Japanese patients with solar urticaria, although the mechanism of action of the IS has not been elucidated. METHODS: Because an intradermal injection of action spectrum (AS)-irradiated serum in a case of solar urticaria induced a wheal response, we studied the responsiveness of the intradermal injection after an IS irradiation. RESULTS: An AS in this patient was composed of visible light shorter than 500 nm, while an IS was composed of visible light longer than 530 nm. When the IS was exposed immediately after the AS irradiation, the wheal response was inhibited. However, when the IS was exposed before the AS irradiation, the wheal response was not inhibited. An intradermal injection of her serum produced no reaction, whereas an intradermal injection of her serum pre irradiated with visible light induced a wheal flare response. Further examination revealed that the in vivo wheal-inducing activity of her serum irradiated with visible light could be attenuated by post-IS irradiation at the injection site, while the wheal-inducing activity of her visible light-irradiated serum was not inhibited by irradiation of the activated serum with the IS. The wheal-flare response induced by compound 48/80 and histamine was not altered by IS irradiation at the site of skin tests. CONCLUSION: These findings indicate that photoallergens in the patient's serum that are activated by visible light irradiation are responsible for the development of her symptoms and that the IS may suppress the wheal response by inhibiting the binding of the photoallergens to mast cells, not by inactivating the photoallergens and stabilizing mast cells. PMID- 16719866 TI - Treatment of localized vitiligo with targeted broadband UVB phototherapy: a pilot study. AB - BACKGROUND: Narrowband ultraviolet B (UVB) phototherapy and 308 nm excimer laser have produced encouraging therapeutic results for vitiligo. Repigmentation of various degrees was obtained in different studies. MATERIALS AND METHODS: Twenty nine vitiliginous lesions from six patients were treated with targeted, broadband UV-B phototherapy. UV fluences were started at 50% of the minimal erythema dose, then increased gradually. Treatments were carried out twice weekly for 12 weeks. RESULTS: Some degree of repigmentation occurred in all subjects. Responses varied among the different anatomic locations, with acral lesions achieving the least improvement. Onset of repigmentation was as early as 3 weeks of treatment in some subjects. Treatments were well tolerated, with only minimal erythema and hyperpigmentation. LIMITATIONS: This study was carried out in a smaller number of patients with skin types III and IV. The irradiation device was a broadband UVB device, and thus the results may not be similar to those obtained from a more monochromatic system such as an excimer laser. CONCLUSIONS: Targeted broadband UVB is an efficacious and safe modality for the treatment of localized vitiligo. PMID- 16719867 TI - Effects of some emollients on the transmission of ultraviolet. AB - BACKGROUND/PURPOSE: Various topical agents which can be used in combination with phototherapy may have blocking or enhancing effects. In this in vivo study, the effects of topical petrolatum, basis cream, glycerine and olive oil on the transmission of ultraviolet B (UVB) were investigated. METHODS: A phototest was performed to determine the minimal erythema dose (MED) on 32 volunteers and the test was repeated with white petrolatum, basis cream, glycerine, olive oil and sunscreen (0.3 cm3/25 cm2). The effects of each agent on MED was determined after 24 h. RESULTS: MEDs were increased with the application of white petrolatum and basis cream. However, MEDs were not changed with the application of glycerine and olive oil. CONCLUSION: The application of white petrolatum and basis cream, which have blocking effects, immediately before UVB therapy is not recommended. However, glycerine and olive oil, which does not have any effect, can be used before phototherapy. PMID- 16719868 TI - Pheomelanin and eumelanin in human skin determined by high-performance liquid chromatography and its relation to in vivo reflectance measurements. AB - BACKGROUND/PURPOSE: Clinical experience has shown that red haired, fair-skinned people have an increased frequency of skin cancer and in addition a relatively high content of pheomelanin and low content of eumelanin in their skin. Of these, eumelanin is presumed photoprotective, and pheomelanin phototoxic. Thus, a fast, easy, and non-invasive method for determining skin content of eumelanin and pheomelanin was sought. METHODS: Skin reflectance measurements using a UV Optimize apparatus were performed before taking suction blisters of the same skin area. Each skin sample was freeze dried and submitted to alkaline hydrogen peroxide degradation. Two eumelanin degradation compounds:Pyrrole-2,3 dicarboxylic acid (PDCA) and pyrrole-2,3,5-tricarboxylic acid (PTCA), and two pheomelanin degradation compounds: Thiazole-4,5-dicarboxylic acid (TDCA) and 1,3 thiazole-2,4,5-tricarboxylic acid (TTCA) were determined in a single chromatographic analysis. Each degradation compound was correlated to reflectance measurements of red (660 nm), green (555 nm), and blue (488 nm) light and additionally correlated to the pigmentation % and redness % given by UV-Optimize. RESULTS: Neither of the investigated parameters were significantly correlated to either PDCA or TTCA. Contrarily, a highly significant correlation was found for the eumelanin marker, PTCA (r2=0.79, P<0.0001). The pheomelanin marker, TDCA, was likewise found to be significantly correlated (r2=0.43, P<0.0001). CONCLUSION: Based on the coefficients of determination it was concluded that in vivo spectrophotometric reflectance measurements of human skin were highly useful for determining the eumelanin content of skin and to classify the pheomelanin content of skin non-invasively. This may be used individually and in population-based studies. PMID- 16719869 TI - Minimal heating dose: a novel biological unit to measure infrared irradiation. AB - BACKGROUND AND OBJECTIVE: Infrared (IR) rays, which comprise approximately 40% of the solar radiation which reaches the earth's surface, have received relatively scant attention. As no standard method has yet been agreed upon for the biological evaluation of IR irradiation, the objective of this study is to suggest a new unit for IR irradiation. METHODS: The skin temperature of 38 Korean volunteers was measured after IR irradiation with varying irradiance. RESULTS: Skin temperature after IR irradiation at an irradiance of 2.02 W/cm2 remained unchanged after 652+/-22 s (mean+/-standard error), which corresponds, in this case, to a total radiation dose of IR 1317.3+/-44.84 J/cm2. This quantity was designated as the minimal heating dose (MHD). We also demonstrated that MHD increased with increasing IR irradiance at lower IR irradiance (1.17 and 2.02 W/cm2), whereas it became constant at higher irradiance (2.87 and 3.22 W/cm2). No statistically significant correlations were detected between MHD and volunteers' ages, erythema index, or melanin index. CONCLUSION: We propose 'MHD' as a biological unit for the measurement of IR irradiation. PMID- 16719870 TI - Photodynamic action of red light for treatment of erythrasma: preliminary results. AB - BACKGROUND: Erythrasma is a superficial cutaneous infection caused by Corynebacterium minutissimum and is characterized by fluorescence under Wood's light (UV) because of the presence of porphyrins. These molecules are photosensitizing and we propose to assess efficacy of red light that activates porphyrins (photodynamic reaction) in treatment of this pathology. OBJECTIVES: Assessment of effects of photodynamic action of red light for treatment of erythrasma without exogenous photosensitizing molecules. METHODS: Thirteen patients with erythrasma were treated by one illumination (80 J/cm2) by red light (broad band, peak at 635 nm) without exogenous photosensitizing molecules. Disappearance or reduction of extent of lesions were observed 2 weeks later. If lesions were still present, a second irradiation was conducted with the same method. RESULTS: Preliminary results are presented. As a result of red light irradiation, we noticed a complete recovery for three patients and, in most other cases, reduction of extent of lesions (mean: -29% after one session). The treatment was well tolerated. CONCLUSION: We report first cases of photodynamic treatment of erythrasma. There are other reports of clinical applications of antimicrobial action of photodynamic therapy in dermatology (acne vulgaris, leishmaniasis, warts, etc.). But there are few applications without addition of exogenous photosensitizing agent. The originality and interest of our study is to use spontaneous presence of porphyrins in the lesions. This technique seems to be an interesting alternative, inexpensive and easy, for the treatment of this localized infection. But an optimal method is still to be determined to improve efficacy. PMID- 16719871 TI - Evaluation of the effects of NB-UVB in both segmental and non-segmental vitiligo affecting different body sites. AB - BACKGROUND: Narrow band (NB)-UVB has been used in the treatment of vitiligo for years but statistical evaluation of the clinical response in both segmental and non-segmental vitiligo patients has yet to be assessed. OBJECTIVES: Statistical evaluation of the clinical response of vitiligo patients to NB-UVB in both segmental and non-segmental types affecting different body sites. METHODS: This study included 150 patients with vitiligo either segmental (10%) or non-segmental (90%). NB-UVB therapy was given twice weekly till reaching our end point of 100% re-pigmentation or a cut point in unresponsive cases. Evaluation of the percentage of re-pigmentation was performed by total body photography and planimetry every 8 weeks. RESULTS: The overall response to therapy in the non segmental vitiligo group demonstrated that 48% of the patients showed marked response, 27% showed moderate response and 25% showed mild response after UVB therapy. The patients showed marked response in 76.3% in face lesions, 41.9% in trunk lesions and 37.6% in limbs lesions. None of the patients in the acral areas achieved marked response. The mean duration of therapy was 7.8 months. Moreover, the results demonstrated that the earlier the patient was treated, the better the response was especially for lesions on the face, trunk and limbs. On the other hand, in the segmental vitiligo group, patients showed no more than mild response to NB-UVB whatever the site of the lesion was. No side effects were encountered with NB-UVB therapy except for aggravation of the disease in two cases and erythema in one patient who was an outdoor worker and was skin type II. CONCLUSION: The type of vitiligo, the affected anatomical area and the disease duration are important factors that influence potential re-pigmentation. PMID- 16719872 TI - Ultraviolet A-1 as a treatment for ulcerative lichen planus of the feet. AB - Ulcerative lichen planus of the feet is a rare variant of lichen planus characterized by the constellation of a chronic bullous and ulcerative eruption limited to the feet, atrophy of involved skin and nails, and cicatricial alopecia. Only split-thickness skin grafting has here to been documented to be effective. We here report a case with ulcerative lichen planus of the feet successfully treated with medium-dose ultraviolet A-1. PMID- 16719873 TI - Haemorrhagic polymorphic light eruption: two cases of a rare variant. AB - Polymorphic light eruption is a common photosensitivity disorder of unknown aetiology, that usually presents in the spring or summer months as an intermittent non-scarring, itchy erythematous, papulo-vesicular eruption. We present two cases of haemorrhagic polymorphic light eruption, a rare variant of this condition of which there are no case reports in the literature. PMID- 16719874 TI - 308-nm excimer laser for the treatment of lymphomatoid papulosis and stage IA mycosis fungoides. PMID- 16719875 TI - New pediatric resuscitation guidelines: new evidence or new ideas? PMID- 16719876 TI - Anesthesia management of familial dysautonomia. AB - Familial dysautonomia (FD) is an autosomal recessive inherited disorder, predominantly affecting the Ashkenazi Jewish population that is characterized by sensory and autonomic neuropathy. The protean manifestations and perturbations result in high morbidity and mortality. However, as a result of supportive measures and centralized care, survival has improved. As surgical options are increasing to symptomatically treat FD, anesthesiologists need to be familiar with this disorder. Because the Dysautonomia Center at NYU Medical Center is a referral center for FD patients, we have attained considerable anesthetic experience with FD. This article reviews clinical features of FD that could potentially affect anesthetic management and outlines our present practices. PMID- 16719877 TI - Oral sedation with midazolam and diphenhydramine compared with midazolam alone in children undergoing magnetic resonance imaging. AB - BACKGROUND: The purpose of this study was to compare the safety and efficacy of oral midazolam and midazolam-diphenhydramine combination to sedate children undergoing magnetic resonance imaging (MRI). METHODS: We performed a prospective randomized double-blind study in 96 children who were randomly allocated into two groups. Group D received oral diphenhydramine (1.25 mg x kg(-1)) with midazolam (0.5 mg x kg(-1)), and Group P received oral placebo with midazolam (0.5 mg x kg( 1)) alone. Sedation scores, onset and duration of sleep were evaluated. Adverse effects, including hypoxemia, failed sedation, and the return of baseline activity, were documented. RESULTS: Diphenhydramine facilitated an earlier onset of midazolam sedation (P < 0.01), and higher sedation scores (P < 0.01). In children who received midazolam alone, 20 (41%) were inadequately sedated, compared with 9 (18%) children who received midazolam and diphenhydramine combination (P < 0.01). Time to complete recovery was not significantly different between the two groups. CONCLUSIONS: Our study indicates that the combination of oral diphenhydramine with oral midazolam resulted in safe and effective sedation for children undergoing MRI. The use of this combination might be more advantageous compared with midazolam alone, resulting in less sedation failure during MRI. PMID- 16719878 TI - Predicting which children benefit most from parental presence during induction of anesthesia. AB - BACKGROUND: The purpose of this large-scale prospective cohort study (n = 426) was to identify child and parent characteristics that are associated with low anxiety and good compliance during induction of anesthesia when parents are present. METHODS: Outcome variables included child's anxiety and child's compliance during induction of anesthesia. Predictor variables included demographics, temperament, trait (baseline) anxiety, coping style, and locus of control. RESULTS: Results of a linear regression model (overall proportion of variance accounted for equals 39.5%) showed that significant predictors of anxiety during induction of anesthesia while parents are present included: the child's age (DeltaR(2) = 0.315, P = 0.0001), behavior during previous medical visits (DeltaR(2) = 0.025, P = 0.001), child's activity level (DeltaR(2) = 0.016, P = 0.007), parent's state (contextual) anxiety (DeltaR(2) = 0.022, P = 0.001) and parent's locus of control (DeltaR(2) = 0.009, P = 0.036). A linear regression model that was constructed with compliance of the child as the outcome revealed similar findings. CONCLUSIONS: Children who benefit from parental presence are older, had lower levels of activity in their temperament, and had parents who were calmer and who valued preparation and coping skills for medical situations. The practical implications of these findings are discussed. PMID- 16719879 TI - Estimation of the parameters of cardiac function and of blood volume by arterial thermodilution in an infant animal model. AB - BACKGROUND: Experimental studies in adults and in animals have reported that estimation of the intracardiac volumes by arterial thermodilution is a more reliable method of blood volume estimation than pressure measurement. The objective of this study has been to analyze the values of cardiac function and blood volume in an infant animal model using the arterial thermodilution technique. METHODS: A total of 202 measurements of cardiac output were performed by femoral arterial thermodilution in 38 Maryland piglets weighing between 8 and 16 kg, to determine the normal values of blood volume obtained by arterial thermodilution (PiCCO method) in an infant animal model. The following parameters were measured: blood volume [global end-diastolic volume index (GEDVI), total intrathoracic blood index (ITBI), extravascular lung water index (ELWI), systolic volume index (SVI)] and parameters of cardiac and vascular function [systolic volume index (SVI), cardiac function index (CFI), left ventricular contractility (Dp/dtmax), and systemic vascular resistance index) (SVRI)]. RESULTS: The cardiac index, 4.3 +/- 1.2 l x min(-1) x m(2), was within the normal range. The GEDVI, 198 +/- 48.6 ml x m(2), and ITBI, 574.1 +/- 113.4 ml x m(2), were lower than the normal values reported in adults, whereas the ELWI, 16.3 +/- 5.2 ml x kg(-1), was higher. CONCLUSIONS: Intrathoracic and intracardiac volume values obtained by arterial thermodilution are lower than those considered normal in the adult, whereas the extravascular lung water is higher. These values must be taken into account when the PiCCO method is used in small children. PMID- 16719880 TI - Optimally fitted tracheal tubes decrease the probability of postextubation adverse events in children undergoing general anesthesia. AB - BACKGROUND: The air leak test is recommended for assessing the appropriate size of an uncuffed tracheal tube (TT) in children. Our objectives were to determine whether there is a certain threshold air leak value beyond which a higher risk for adverse events after removal of TT can be predicted and to define other risk factors related to extubation. METHODS: We enrolled 234 cases ranging from newborn to 9 years of age requiring tracheal intubation for elective or emergency surgery. General anesthesia was induced by a mask or intravenously. The TT size was calculated using the formula: [age (years)/4] + 4.5. After the induction of anesthesia, the patient's trachea was intubated and the correct position was confirmed. The attending anesthetist assessed the leak pressure. Incidences of adverse events (prolonged or barking cough, obstructed or prolonged inspiration or expiration, subcostal and sternal retractions, arterial desaturation, or laryngospasm) were recorded after removal of TT. RESULTS: Ten patients were excluded from the study. A total of 218 children underwent 224 operations under general anesthesia. Children who had an absent air leak at 25 cmH(2)O pressure had 2.8 times more adverse events during emergence from anesthesia than those with an audible air leak. Adverse events after the removal of TT were 3.7 times more likely to occur in children whose anesthesia was provided by a less experienced anesthesia trainee. CONCLUSIONS: Adverse events after removal of TT were more likely to occur in children with an absent air leak at 25 cmH(2)O pressure and in children whose anesthesia was provided by a less experienced anesthetist. PMID- 16719881 TI - Tramadol vs morphine during adenotonsillectomy for obstructive sleep apnea in children. AB - BACKGROUND: Optimal analgesia for children undergoing adenotonsillectomy for obstructive sleep apnea (OSA) is controversial. Tramadol may represent a superior choice over morphine in this group, with a potential to cause less postoperative sedation and respiratory depression. Optimal perioperative analgesia may allow expensive and time-consuming preoperative work-up and postoperative monitoring to be rationalized. METHODS: Sixty-six children were randomized to receive either perioperative tramadol or morphine in this double blinded, prospective, controlled trial. Postoperative sedation, pain, respiratory events, and vomiting were then compared between groups. RESULTS: There was no significant difference between the two groups in sedation scores 1 h after arrival in recovery (P = 0.24) or at any other time up to 6 h postoperation. There was also no evidence of a difference between the groups in pain scores up to 6 h postoperation. There were fewer episodes of postoperative desaturation (<94%) in the tramadol group up to 3 h postoperation, with 26% fewer episodes in the tramadol group during the second hour postoperation (P = 0.02). Overall, there was a trend toward fewer desaturation episodes in the tramadol group. CONCLUSIONS: Tramadol may be a suitable drug for children undergoing adenotonsillectomy for OSA. Further work is required to investigate this. PMID- 16719883 TI - Convection heating in pediatric general surgery - a comparison of warming alternatives in a mannequin study. AB - BACKGROUND: Numerous methods of patient warming are used to prevent intraoperative hypothermia in children. Commercially available forced air warming blankets are effective, but are single-use items. We tested a custom-designed heat dissipation unit (HDU) against one such commercially available blanket. METHODS: Air temperatures at various points around a mannequin under simulated operating conditions were recorded using thermistors and thermal imaging. The only variable changed was the heating method: a forced air blanket or a customized HDU with two draping techniques - cotton drapes with and without a plastic 'undersheet'. RESULTS: The three methods produced similar temperature increases and plateaux across the 11 thermistor points measured. There were no significant differences between temperatures at 1 h. A plastic sheet did not appear to enhance the effectiveness of the HDU in this study. Thermal imaging photography suggested more uniform heating of the mannequin with the HDU arrangements. CONCLUSIONS: The custom-built HDU compares favorably in our mannequin study with a Bair Hugger forced air warming blanket. As it is reusable, it offers considerable potential savings. PMID- 16719882 TI - The safety and effectiveness of a modified convection heating system for children during anesthesia. AB - BACKGROUND: Convection heating shows most promise in maintaining children's core temperatures under anesthesia. We have previously shown that a modified convection heating technique worked in a mannequin model and sought to establish its safety and effectiveness in a clinical study. METHODS: Children were recruited who were having elective surgery under general anesthesia lasting >90 min. The children were anesthetized and maintained in a room temperature of 21 degrees C. Warming was performed by a 'Bair Hugger' attached to a heat dissipation box, producing turbulent air from multiple outlet holes on its face. A plastic sheet covered the child, was attached to the top of the box, tucked into the sides of the bed and left open at the head end. Temperatures at various sites (air, skin, and core) were continuously monitored using thermistors connected to a datalogger and laptop. Analysis was performed using Excel. RESULTS: The study comprised 40 children ranging in age from 2 days to 12.5 years and weigh 2.5-73 kg. Operations were 'peripheral' (e.g. urethroplasty) lasting 90 min to major laparotomy lasting 590 min. Body surface area uncovered was 5-25%. Skin temperatures rose to a maximum of c. 40 degrees C. Core temperatures rose after a 12-min lag by 0.01-0.04 degrees C x min(-1). In children who became hyperthermic, cooling was readily achieved by turning the heating off and leaving the fan running. CONCLUSIONS: The technique is safe and effective for children throughout the pediatric range. The practice of increasing room temperature above 21 degrees C for elective cases should be abandoned. Continuous monitoring of core temperature is necessary to prevent hyperthermia. PMID- 16719884 TI - Management of a cadaveric orthotopic liver transplantation in a pediatric patient with complex congenital heart disease. AB - Pediatric orthotopic liver transplantations (OLT) are commonly performed nowadays. Two primary reasons for OLT in children are complications from either extrahepatic biliary atresia (EHBA) or inborn errors of metabolism. However, congenital liver disease may be associated with significant other congenital abnormalities. We present a case of a successful OLT in a pediatric patient with a history of EHBA, situs inversus, and complex congenital heart disease. The cardiac anomalies include dextrocardia, absence of the atrial septum (single atrium), single atrioventricular valve (a-v canal), and an incomplete ventricular septum. Prior surgery include a Kasai procedure for EHBA, banding of the proximal main pulmonary artery, and Broviac catheter placement. We present the anesthesia concerns and management for this complicated case. PMID- 16719885 TI - Grand mal seizures: an unusual and puzzling primary presentation of ruptured hepatic hydatid cyst. AB - We report a case of hepatic hydatidosis where the first clinical manifestations, generalized seizures after minor head and abdominal trauma, and delayed anaphylaxis, made the primary diagnosis difficult. Severe anaphylaxis has been reported as initial presentation of quiescent hepatic hydatidosis. In endemic areas, the diagnosis must be carefully ruled out in patients experiencing abrupt anaphylactic shock of uncertain etiology. The occurrence of unexplained vascular collapse after minor abdominal trauma in a patient originating from an endemic area should prompt the diagnosis and urgent treatment should be initiated; firstly emergency management of the anaphylactic shock and later, surgical treatment of the cysts. PMID- 16719886 TI - Acute pancreatitis after anesthesia with propofol in a child with glycogen storage disease type IA. AB - Glycogen storage disorder type 1A (GSD 1A) is an inherited disorder of glycogen metabolism characterized by fasting hypoglycemia, lactic acidosis, hyperuricemia, and hyperlipidemia. These children have a higher risk of developing pancreatitis because of hypertriglyceridemia. Drug-induced pancreatitis accounts for a small proportion of cases of pancreatitis. The mechanism of drug-induced pancreatitis include hypersensitivity, direct toxic injury or indirectly by inducing hypertriglyceridemia. Propofol is often the drug of choice for induction of anesthesia in ambulatory surgical procedures. There are various reports in the literature describing pancreatitis induced by propofol. We present a 4-year-old girl with GSD 1A, who required tonsillectomy and adenoidectomy under general anesthesia. She developed acute pancreatitis in the postoperative period. Propofol was used as a general anesthetic and the postoperative incidence of pancreatitis is discussed. PMID- 16719887 TI - Anesthesia in a child with fibrodysplasia ossificans progressiva. AB - We describe the anesthesia management of a 12-year-old girl, diagnosed with fibrodysplasia ossificans progressiva (FOP), who presented with a submandibular abscess. FOP is a rare, inherited disorder with heterotopic bone formation and progressive musculoskeletal disability. This disability ultimately confines patients to a wheelchair. Minor trauma following dental treatment may lead to ankylosis of the jaw. Subsequent to this disability, which resulted in poor dental hygiene, our patient developed a dental abscess. This spread along the mandibular margin and under the tongue. She presented with an impending airway compromise in an already difficult situation. The options for airway management in a child with limited mouth opening are discussed. PMID- 16719888 TI - Glottic scar bands following intubation. AB - We present two uncommon cases of abnormalities of the pediatric airway, which may present in the first instance to the anesthetist. Glottic scar bands are a result of intubation trauma and are a treatable cause of voice abnormalities and sometimes respiratory distress. PMID- 16719889 TI - Epidural analgesia in the surgery of congenital tracheal stenosis: slide tracheoplasty on cardiopulmonary bypass. AB - Epidural analgesia in children is highly effective and safe; however, it has not enjoyed great popularity in surgery that requires cardiopulmonary bypass. A major concern is the possibility of damage to blood vessels with the epidural needle or catheter and epidural hematoma formation. There seems to be a low incidence of epidural hematoma if certain guidelines are followed, so that in children, epidural analgesia can be used in selected patients, with safety, when surgical repair requires cardiopulmonary bypass. Epidural morphine has been used for clinical pain relief in pediatric cardiac surgery. Improved pulmonary function, suppressed hormonal and metabolic stress responses, easy early tracheal extubation, and good analgesia and sedation that allows neurological examination to alert any possibles hidden complications, are the advantages. A dedicated medical team is essential in the perioperative management to achieve maximum benefit for these patients. PMID- 16719890 TI - Unexpected hypernatremia during pulmonary resection in a 7-year-old child with hydatid disease. PMID- 16719891 TI - Anesthetic management with laryngeal mask in a child with Brachmann-de Lange syndrome. PMID- 16719892 TI - Lumbar epidural anesthesia for the child with Escobar syndrome. PMID- 16719893 TI - Propofol-induced bradycardia in a pediatric patient undergoing chemotherapy with the known cardiotoxic agent cytarabin. PMID- 16719895 TI - Low flow and temperature. PMID- 16719896 TI - An innocent perioperative position, an unexpected postoperative complication: bilateral foot drop. PMID- 16719897 TI - Intraoperative anaphylaxis to a chlorhexdine polymer in childhood. PMID- 16719899 TI - Negative impact of chronic obstructive pulmonary disease on the health-related quality of life of patients. Results of the EPIDEPOC study. AB - BACKGROUND: COPD is currently the fourth cause of morbidity and mortality in the developed world. Patients with COPD experience a progressive deterioration and disability, which lead to a worsening in their health-related quality of life (HRQoL). The aim of this work is to assess the Health-Related Quality of Life (HRQoL) of patients with stable COPD followed in primary care and to identify possible predictors of disease. METHODS: It is a multicenter, epidemiological, observational, descriptive study. Subjects of both sexes, older than 40 years and diagnosed of COPD at least 12 months before starting the study were included. Sociodemographic data, severity of disease, comorbidity, and use of health resources in the previous 12 months were collected. All patients were administered a generic quality-of-life questionnaire, the SF-12, that enables to calculate two scores, the physical (PCS-12) and the mental (MCS-12) component summary scores. RESULTS: 10,711 patients were evaluated (75.6% men, 24.4% women), with a mean age of 67.1 years (SD 9.66). The mean value of FEV1 was 35.9 +/- 10.0%. Mean PCS-12 and MCS-12 scores were 36.0 +/- 9.9 and 48.3 +/- 10.9, respectively. Compared to the reference population, patients with COPD had a reduction of PCS-12, even in mild stages of the disease. The correlation with FEV1 was higher for PCS-12 (r = 0.38) than for MCS-12 (r = 0.12). Predictors for both HRQoL components were sex, FEV1, use of oxygen therapy, and number of visits to emergency rooms and hospital admissions. Other independent predictors of PCS 12 were age, body mass index and educational level. CONCLUSION: Patients with stable COPD show a reduction of their HRQoL, even in mild stages of the disease. The factors determining the HRQoL include sex, FEV1, use of oxygen therapy, and number of visits to emergency rooms and hospital admissions. PMID- 16719900 TI - Primary care staff's views and experiences related to routinely advising patients about physical activity. A questionnaire survey. AB - BACKGROUND: United Kingdom public health policy has recently re-emphasised the role of primary health care professionals in tackling increasing levels of physical inactivity within the general population. However, little is known about the impact that this has had in practice. This study explores Scottish primary care staff's knowledge, attitudes and experiences associated with advising patients about physical activity during routine consultations. METHODS: A cross sectional questionnaire survey of general practitioners (or family physicians), practice nurses and health visitors based in four health regions was conducted during 2004. The main outcome measures included: (i) health professionals' knowledge of the current physical activity recommendations; (ii) practice related to routine physical activity advising; and (iii) associated attitudes. RESULTS: Questionnaires were returned by 757 primary care staff (response rate 54%). Confidence and enthusiasm for giving advice was generally high, but knowledge of current physical activity recommendations was low. In general, respondents indicated that they routinely discuss and advise patients about physical activity regardless of the presenting condition. Health visitors and practice nurses were more likely than general practitioners to offer routine advice. Lack of time and resources were more likely to be reported as barriers to routine advising by general practitioners than other professional groups. However, health visitors and practice nurses were also more likely than general practitioners to believe that patients would follow their physical activity advice giving. CONCLUSION: If primary health care staff are to be fully motivated and effective in encouraging and supporting the general population to become more physically active, policymakers and health professionals need to engage in efforts to: (1) improve knowledge of current physical activity recommendations and population trends amongst frontline primary care staff; and (2) consider the development of tools to support individual assessment and advice giving to suit individual circumstances. Despite the fact that this study found that system barriers to routine advising were less of a problem than other previous research has indicated, this issue still remains a challenge. PMID- 16719901 TI - Pre-natal and post-natal exposure to respiratory infection and atopic diseases development: a historical cohort study. AB - BACKGROUND: According to the hygiene hypothesis, infections in early life protect from allergic diseases. However, in earlier studies surrogate measures of infection rather than clinical infections were associated with decreased frequencies of atopic diseases. Exposure to infection indicating sub-clinical infection rather than clinical infection might protect from atopic diseases. OBJECTIVE: to investigate whether exposure to acute respiratory infections within pregnancy and the first year of life is associated with atopic conditions at age 5-14 years and to explore when within pregnancy and the first year of life this exposure is most likely to be protective. METHODS: Historical cohort study: Population level data on acute respiratory infections from the routine reporting system of the former German Democratic Republic were linked with individual data from consecutive surveys on atopic diseases in the same region (n = 4672). Statistical analyses included multivariate logistic regression analysis and polynomial distributed lag models. RESULTS: High exposure to acute respiratory infection between pregnancy and age one year was associated with overall reduced odds of asthma, eczema, hay fever, atopic sensitization and total IgE. Exposure in the first 9 months of life showed the most pronounced effect. Adjusted odds ratio's for asthma, hay fever, inhalant sensitization and total IgE were statistical significantly reduced up to around half. CONCLUSION: Exposure to respiratory infection (most likely indicating sub-clinical infection) within pregnancy and the first year of life may be protective in atopic diseases development. The post-natal period thereby seems to be particularly important. PMID- 16719902 TI - HIV and pre-neoplastic and neoplastic lesions of the cervix in South Africa: a case-control study. AB - BACKGROUND: Cervical cancer and infection with human immunodeficiency virus (HIV) are both major public health problems in South Africa. The aim of this study was to determine the risk of cervical pre-cancer and cancer among HIV positive women in South Africa. METHODS: Data were derived from a case-control study that examined the association between hormonal contraceptives and invasive cervical cancer. The study was conducted in the Western Cape (South Africa), from January 1998 to December 2001. There were 486 women with invasive cervical cancer, 103 control women with atypical squamous cells of undetermined significance (ASCUS), 53 with low-grade squamous intraepithelial lesions (LSIL), 50 with high-grade squamous intraepithelial lesions (HSIL) and 1159 with normal cytology. Odds ratios (OR) and 95% confidence intervals (CI) were calculated using multiple logistic regression. RESULTS: The adjusted odds ratios associated with HIV infection were: 4.4 [95% CI (2.3 - 8.4) for ASCUS, 7.4 (3.5 - 15.7) for LSIL, 5.8 (2.4 - 13.6) for HSIL and 1.17 (0.75 - 1.85) for invasive cervical cancer. HIV positive women were nearly 5 times more likely to have high-risk human papillomavirus infection (HR-HPV) present compared to HIV negative women [OR 4.6 (95 % CI 2.8 - 7.5)]. Women infected with both HIV and high-risk HPV had a more than 40 fold higher risk of SIL than women infected with neither of these viruses. CONCLUSION: HIV positive women were at an increased risk of cervical pre cancer, but did not demonstrate an excess risk of invasive cervical cancer. An interaction between HIV and HR-HPV infection was demonstrated. Our findings underscore the importance of developing locally relevant screening and management guidelines for HIV positive women in South Africa. PMID- 16719903 TI - Actin polymerisation at the cytoplasmic face of eukaryotic nuclei. AB - BACKGROUND: There exists abundant molecular and ultra-structural evidence to suggest that cytoplasmic actin can physically interact with the nuclear envelope (NE) membrane system. However, this interaction has yet to be characterised in living interphase cells. RESULTS: Using a fluorescent conjugate of the actin binding drug cytochalasin D (CD-BODIPY) we provide evidence that polymerising actin accumulates in vicinity to the NE. In addition, both transiently expressed fluorescent actin and cytoplasmic micro-injection of fluorescent actin resulted in accumulation of actin at the NE-membrane. Consistent with the idea that the cytoplasmic phase of NE-membranes can support this novel pool of perinuclear actin polymerisation we show that isolated, intact, differentiated primary hepatocyte nuclei support actin polymerisation in vitro. Further this phenomenon was inhibited by treatments hindering steric access to outer-nuclear-membrane proteins (e.g. wheat germ agglutinin, anti-nesprin and anti-nucleoporin antibodies). CONCLUSION: We conclude that actin polymerisation occurs around interphase nuclei of living cells at the cytoplasmic phase of NE-membranes. PMID- 16719904 TI - Texture discrimination and multi-unit recording in the rat vibrissal nerve. AB - BACKGROUND: Rats distinguish objects differing in surface texture by actively moving their vibrissae. In this paper we characterized some aspects of texture sensing in anesthetized rats during active touch. We analyzed the multifiber discharge from a deep vibrissal nerve when the vibrissa sweeps materials (wood, metal, acrylic, sandpaper) having different textures. We polished these surfaces with sandpaper (P1000) to obtain close degrees of roughness and we induced vibrissal movement with two-branch facial nerve stimulation. We also consider the change in pressure against the vibrissa as a way to improve the tactile information acquisition. The signals were compared with a reference signal (control)--vibrissa sweeping the air--and were analyzed with the Root Mean Square (RMS) and the Power Spectrum Density (PSD). RESULTS: We extracted the information about texture discrimination hidden in the population activity of one vibrissa innervation, using the RMS values and the PSD. The pressure level 3 produced the best differentiation for RMS values and it could represent the "optimum" vibrissal pressure for texture discrimination. The frequency analysis (PSD) provided information only at low-pressure levels and showed that the differences are not related to the roughness of the materials but could be related to other texture parameters. CONCLUSION: Our results suggest that the physical properties of different materials could be transduced by the trigeminal sensory system of rats, as are shown by amplitude and frequency changes. Likewise, varying the pressure could represent a behavioral strategy that improves the information acquisition for texture discrimination. PMID- 16719906 TI - Vibratory stimulation increase the electro-cutaneous sensory detection and pain thresholds in women but not in men. AB - BACKGROUND: Vibratory stimulation is a potential method for the treatment of pain. METHODS: The effect of vibration on the forearm on detection (DT) and pain thresholds (PT) induced by electro-cutaneous stimulation were investigated in healthy male and female volunteers. RESULTS: Women have lower baseline detection and pain thresholds as compared to men. Furthermore, women but not men report increased detection and pain thresholds after vibratory stimulation. CONCLUSION: Our findings indicate the potential usefulness of vibratory stimulation for pain treatment, and that gender differences should be considered in future evaluation of the method. PMID- 16719905 TI - Association study of genetic variants of pro-inflammatory chemokine and cytokine genes in systemic lupus erythematosus. AB - BACKGROUND: Several lines of evidence suggest that chemokines and cytokines play an important role in the inflammatory development and progression of systemic lupus erythematosus. The aim of this study was to evaluate the relevance of functional genetic variations of RANTES, IL-8, IL-1alpha, and MCP-1 for systemic lupus erythematosus. METHODS: The study was conducted on 500 SLE patients and 481 ethnically matched healthy controls. Genotyping of polymorphisms in the RANTES, IL-8, IL-1alpha, and MCP-1 genes were performed using a real-time polymerase chain reaction (PCR) system with pre-developed TaqMan allelic discrimination assay. RESULTS: No significant differences between SLE patients and healthy controls were observed when comparing genotype, allele or haplotype frequencies of the RANTES, IL-8, IL-1alpha, and MCP-1 polymorphisms. In addition, no evidence for association with clinical sub-features of SLE was found. CONCLUSION: These results suggest that the tested functional variation of RANTES, IL-8, IL-1alpha, and MCP-1 genes do not confer a relevant role in the susceptibility or severity of SLE in the Spanish population. PMID- 16719907 TI - Comparative genomic mapping of the bovine Fragile Histidine Triad (FHIT) tumour suppressor gene: characterization of a 2 Mb BAC contig covering the locus, complete annotation of the gene, analysis of cDNA and of physiological expression profiles. AB - BACKGROUND: The Fragile Histidine Triad gene (FHIT) is an oncosuppressor implicated in many human cancers, including vesical tumors. FHIT is frequently hit by deletions caused by fragility at FRA3B, the most active of human common fragile sites, where FHIT lays. Vesical tumors affect also cattle, including animals grazing in the wild on bracken fern; compounds released by the fern are known to induce chromosome fragility and may trigger cancer with the interplay of latent Papilloma virus. RESULTS: The bovine FHIT was characterized by assembling a contig of 78 BACs. Sequence tags were designed on human exons and introns and used directly to select bovine BACs, or compared with sequence data in the bovine genome database or in the trace archive of the bovine genome sequencing project, and adapted before use. FHIT is split in ten exons like in man, with exons 5 to 9 coding for a 149 amino acids protein. VISTA global alignments between bovine genomic contigs retrieved from the bovine genome database and the human FHIT region were performed. Conservation was extremely high over a 2 Mb region spanning the whole FHIT locus, including the size of introns. Thus, the bovine FHIT covers about 1.6 Mb compared to 1.5 Mb in man. Expression was analyzed by RT PCR and Northern blot, and was found to be ubiquitous. Four cDNA isoforms were isolated and sequenced, that originate from an alternative usage of three variants of exon 4, revealing a size very close to the major human FHIT cDNAs. CONCLUSION: A comparative genomic approach allowed to assemble a contig of 78 BACs and to completely annotate a 1.6 Mb region spanning the bovine FHIT gene. The findings confirmed the very high level of conservation between human and bovine genomes and the importance of comparative mapping to speed the annotation process of the recently sequenced bovine genome. The detailed knowledge of the genomic FHIT region will allow to study the role of FHIT in bovine cancerogenesis, especially of vesical papillomavirus-associated cancers of the urinary bladder, and will be the basis to define the molecular structure of the bovine homologue of FRA3B, the major common fragile site of the human genome. PMID- 16719908 TI - Monthly intravenous methylprednisolone in relapsing-remitting multiple sclerosis reduction of enhancing lesions, T2 lesion volume and plasma prolactin concentrations. AB - BACKGROUND: Intravenous methylprednisolone (IV-MP) is an established treatment for multiple sclerosis (MS) relapses, accompanied by rapid, though transient reduction of gadolinium enhancing (Gd+) lesions on brain MRI. Intermittent IV-MP, alone or with immunomodulators, has been suggested but insufficiently studied as a strategy to prevent relapses. METHODS: In an open, single-cross-over study, nine patients with relapsing-remitting MS (RR-MS) underwent cranial Gd-MRI once monthly for twelve months. From month six on, they received a single i.v. infusion of 500 mg methylprednisolone (and oral tapering for three days) after the MRI. Primary outcome measure was the mean number of Gd+ lesions during treatment vs. baseline periods; T2 lesion volume and monthly plasma concentrations of cortisol, ACTH and prolactin were secondary outcome measures. Safety was assessed clinically, by routine laboratory and bone mineral density measurements. Soluble immune parameters (sTNF-RI, sTNF-RII, IL1-ra and sVCAM-1) and neuroendocrine tests (ACTH test, combined dexamethasone/CRH test) were additionally analyzed. RESULTS: Comparing treatment to baseline periods, the number of Gd+ lesions/scan was reduced in eight of the nine patients, by a median of 43.8% (p = 0.013, Wilcoxon). In comparison, a pooled dataset of 83 untreated RR-MS patients from several studies, selected by the same clinical and MRI criteria, showed a non-significant decrease by a median of 14% (p = 0.32). T2 lesion volume decreased by 21% during treatment (p = 0.001). Monthly plasma prolactin showed a parallel decline (p = 0.027), with significant cross correlation with the number of Gd+ lesions. Other hormones and immune system variables were unchanged, as were ACTH test and dexamethasone-CRH test. Treatment was well tolerated; routine laboratory and bone mineral density were unchanged. CONCLUSION: Monthly IV-MP reduces inflammatory activity and T2 lesion volume in RR-MS. PMID- 16719909 TI - Topiramate in the treatment of compulsive sexual behavior: case report. AB - BACKGROUND: Among the multiple mechanisms of action of topiramate, AMPA/kainate antagonism may be particularly interesting for the treatment of disorders characterized by conditioned cognitive and behavioral cue reactivity. CASE PRESENTATION: We report the case of a patient consulting primarily for obesity and cue triggered snacking, who responded well on topiramate at doses up to 50 mg. Coincidentally he reported on an improvement of compulsive nonparaphilic sexual behaviors (consumption of prostitution), which was also strongly triggered by environmental cues. Both addictive behaviors (snacking and consumption of prostitution) reoccurred after discontinuation of topiramate and again responded reintroduction of the drug. CONCLUSION: The present case report of topiramate's effect on comorbid obesity and nonparaphilic addiction could be interpreted as a further indication that topiramate acts on the common pathway underlying conditioned behaviors and seems to be a treatment of behavioral disorders associated with environmental cues. PMID- 16719911 TI - Care seeking behaviour for childhood illness--a questionnaire survey in western Nepal. AB - BACKGROUND: The World Health Organization estimates that seeking prompt and appropriate care could reduce child deaths due to acute respiratory infections by 20%. The purpose of our study was to assess care seeking behaviour of the mothers during childhood illness and to determine the predictors of mother's care seeking behaviour. METHODS: A cross-sectional survey was conducted in the immunization clinics of Pokhara city, Kaski district, western Nepal. A trained health worker interviewed the mothers of children suffering from illness during the preceding 15 days. RESULTS: A total of 292 mothers were interviewed. Pharmacies (46.2%) were the most common facilities where care was sought followed by allopathic medical practitioners (26.4%). No care was sought for 8 (2.7%) children and 26 (8.9%) children received traditional/home remedies. 'Appropriate', 'prompt' and 'appropriate and prompt' care was sought by 77 (26.4%), 166 (56.8%) and 33 (11.3%) mothers respectively. The mothers were aware of fever (51%), child becoming sicker (45.2%) and drinking poorly (42.5%) as the danger signs of childhood illness. By multiple logistic regression analysis total family income, number of symptoms, mothers' education and perceived severity of illness were the predictors of care seeking behaviour. CONCLUSION: The results of the present study show that the mothers were more likely to seek care when they perceived the illness as 'serious'. Poor maternal knowledge of danger signs of childhood illness warrants the need for a complementary introduction of community-based Integrated Management of Childhood Illness programmes to improve family's care seeking behaviour and their ability to recognize danger signs of childhood illness. Socioeconomic development of the urban poor may overcome their financial constraints to seek 'appropriate' and 'prompt' care during the childhood illness. PMID- 16719910 TI - Expression of carbonic anhydrases IX and XII during mouse embryonic development. AB - BACKGROUND: Of the thirteen active carbonic anhydrase (CA) isozymes, CA IX and XII have been linked to carcinogenesis. It has been suggested that these membrane bound CAs participate in cancer cell invasion, which is facilitated by an acidic tumor cell environment. Since active cell migration is a characteristic feature of embryonic development, we set out to explore whether these isozymes are expressed in mouse embryos of different ages. The studies were focused on organogenesis stage. RESULTS: Immunohistochemistry demonstrated that both CA IX and XII are present in several tissues of the developing mouse embryo during organogenesis. Staining for CA IX revealed a relatively wide distribution pattern with moderate signals in the brain, lung, pancreas and liver and weak signals in the kidney and stomach. The expression pattern of CA XII in the embryonic tissues was also relatively broad, although the intensity of immunostaining was weak in most tissues. The CA XII-positive tissues included the brain, where the most prominent staining was seen in the choroid plexus, and the stomach, pancreas, liver and kidney. CONCLUSION: Membrane-bound CA isozymes IX and XII are expressed in various tissues during mouse organogenesis. These enzymes may regulate ion and pH homeostasis within the developing embryo. PMID- 16719912 TI - Patterns of smoking behavior among physicians in Yerevan, Armenia. AB - BACKGROUND: Physicians can play an important role in smoking prevention and control. This study will identify smoking prevalence among physicians in Yerevan, Armenia. It will also explore how the smoking behaviors of physicians, their perceived ability to influence patient smoking behavior, and their knowledge about health outcomes related to smoking influence their interaction with patients. METHODS: A cross-sectional, self-administered, anonymous survey was conducted in July, 2004, among 12 healthcare facilities in Yerevan. Analyses are based on responses from 240 physicians, representing a 70% response rate. RESULTS: The percentage of current smokers was significantly higher in men than women (48.5% vs. 12.8% regular and 6.8% vs. 4.5% occasional). Among current smokers, 52.7% of men compared with 13.0% of women had previously smoked in the presence of patients. Only 35.3% felt well prepared to assist patients to quit smoking. Physicians who smoke are less likely to ask their patients about their smoking behavior or believe their example is likely to influence their patients. Level of perceived preparedness to assist patients to quit smoking was positively associated with knowledge about known health risks associated with smoking. CONCLUSION: Smoking prevalence is high among physicians in the 12 healthcare facilities in Yerevan, and a large percentage of physician smoke in the presence of their patients. Physician smoking behavior and knowledge of smoking related health outcomes in Yerevan influences whether they counsel patients regarding smoking. PMID- 16719913 TI - Evaluation of the utilization of the preanaesthetic clinics in a university teaching hospital. AB - BACKGROUND: Dedicated out-patient preanaesthetic clinics are relatively recent phenomenon and information is sparse from developing world. This study attempted to evaluate the utilization of adult and paediatric preanaesthetic clinics and its impact on the cancellations of surgery in Trinidad. METHODS: All patients scheduled to have elective surgery during the period of twelve weeks were enrolled for prospective collection of data including demographics, the admitting diagnoses, surgical procedure, category of surgery and specialty, and the patients' attendance to preanaesthetic clinics. Cancellations on the day of surgery along with reasons were recorded. The difference between patients who attended and did not attend the clinic was analysed. RESULTS: Of 424 patients scheduled for procedures during the study period, 213 were adults and 211 were children. Overall 39% of adults and 46% of the children scheduled for surgery had previously attended the preanaesthetic clinic. Among adults, general surgery patients were the largest majority to attend the preanaesthetic clinic. The paediatric preanaesthetic clinic was mostly utilized by paediatric general surgery. Overall 30% of procedures in adults and 26% of those in children were cancelled. There was a statistically significant difference in cancellations between patients who attended and did not attend the preanaesthetic clinic (p = 0.004). There was a 52% more chance of the procedure getting cancelled if the patient did not attend the clinic. CONCLUSION: The study highlights the inadequate use of the preanaesthetic clinics and the impact of the clinics on last-minute cancellations. PMID- 16719914 TI - Trauma induces apoptosis in human thoracolumbar intervertebral discs. AB - BACKGROUND: Vertebral fractures resulting from high energy trauma often comprise the risk of posttraumatic degenerative changes in the affected intervertebral discs (IVD). Particularly in conservatively treated patients, or in cases after implant removal of an exclusively posterior stabilization, consecutive disc degeneration and the associated functional losing of the spinal segment clearly represent detrimental treatment results. In this regard, apoptosis of IVD cells has been suggested to be involved in the critical changes of the extracellular matrix. METHODS: To investigate whether fractures of the vertebrae induce apoptosis in the affected IVD, disc tissue from patients (n = 17) undergoing open reduction and internal fixation of thoracolumbar spine fractures were analysed in regards to caspase activity, apoptosis-receptor expression levels and gene expression of apoptosis-regulating proteins such as Bax and Bcl-2. Healthy IVD tissue (n = 3) obtained from patients undergoing surgical resection of adjacent vertebrae were used as control samples. RESULTS: In contrast to healthy control IVD tissues, samples from traumatic thoracolumbar IVD showed positive TUNEL staining and a significant increase of caspase-3/7 activity. Interestingly, analyses of the initiator caspase-8 and -9 revealed significantly increased activation levels compared to control values, suggesting the coexistent activation of both the extrinsic (receptor-mediated) and intrinsic (mitochondria mediated) apoptosis pathway. Accordingly, expression levels of the Fas receptor (FasR) mRNA were significantly increased. Although the TNF receptor I (TNFR I) was only slightly upregulated, corresponding TNFalpha from trauma IVD presented significantly increased mRNA expression values. Furthermore, traumatic IVD cells demonstrated significantly reduced expression of the mitochondria-bound anti apoptotic Bcl-2, thereby maintaining baseline transcriptional levels of the pro apoptotic Bax protein when compared to control IVD cells. CONCLUSION: Our data suggest that thoracolumbar fractures induce early caspase-dependent apoptosis in IVD cells of the affected intervertebral disc, in part, by downregulation of the anti-apoptotic protein Bcl-2 (intrinsic apoptosis pathway), as well as signalling via the death receptor complex (TNFR I and FasR). PMID- 16719915 TI - The mitochondrial lineage U8a reveals a Paleolithic settlement in the Basque country. AB - BACKGROUND: It is customary, in population genetics studies, to consider Basques as the direct descendants of the Paleolithic Europeans. However, until now there has been no irrefutable genetic proof to support this supposition. Even studies based on mitochondrial DNA (mtDNA), an ideal molecule for constructing datable maternal genealogies, have failed to achieve this. It could be that incoming gene flow has replaced the Basque ancient lineages but it could also be that these lineages have not been detected due to a lack of resolution of the Basque mtDNA genealogies. To assess this possibility we analyzed here the mtDNA of a large sample of autochthonous Basques using mtDNA genomic sequencing for those lineages that could not be unequivocally classified by diagnostic RFLP analysis and control region (HVSI and HVSII) sequencing. RESULTS: We show that Basques have the most ancestral phylogeny in Europe for the rare mitochondrial subhaplogroup U8a. Divergence times situate the Basque origin of this lineage in the Upper Palaeolithic. Most probably, their primitive founders came from West Asia. The lack of U8a lineages in Africa points to an European and not a North African route of entrance. Phylogeographic analysis suggest that U8a had two expansion periods in Europe, the first, from a south-western area including the Iberian peninsula and Mediterranean France before 30,000 years ago, and the second, from Central Europe around 15,000-10,000 years ago. CONCLUSION: It has been demonstrated, for the first time, that Basques show the oldest lineages in Europe for subhaplogroup U8a. Coalescence times for these lineages suggest their presence in the Basque country since the Upper Paleolithic. The European U8 phylogeography is congruent with the supposition that Basques could have participated in demographic re-expansions to repopulate central Europe in the last interglacial periods. PMID- 16719916 TI - Identification of novel functional sequence variants in the gene for peptidase inhibitor 3. AB - BACKGROUND: Peptidase inhibitor 3 (PI3) inhibits neutrophil elastase and proteinase-3, and has a potential role in skin and lung diseases as well as in cancer. Genome-wide expression profiling of chorioamniotic membranes revealed decreased expression of PI3 in women with preterm premature rupture of membranes. To elucidate the molecular mechanisms contributing to the decreased expression in amniotic membranes, the PI3 gene was searched for sequence variations and the functional significance of the identified promoter variants was studied. METHODS: Single nucleotide polymorphisms (SNPs) were identified by direct sequencing of PCR products spanning a region from 1,173 bp upstream to 1,266 bp downstream of the translation start site. Fourteen SNPs were genotyped from 112 and nine SNPs from 24 unrelated individuals. Putative transcription factor binding sites as detected by in silico search were verified by electrophoretic mobility shift assay (EMSA) using nuclear extract from Hela and amnion cell nuclear extract. Deviation from Hardy-Weinberg equilibrium (HWE) was tested by chi2 goodness-of fit test. Haplotypes were estimated using expectation maximization (EM) algorithm. RESULTS: Twenty-three sequence variations were identified by direct sequencing of polymerase chain reaction (PCR) products covering 2,439 nt of the PI3 gene (-1,173 nt of promoter sequences and all three exons). Analysis of 112 unrelated individuals showed that 20 variants had minor allele frequencies (MAF) ranging from 0.02 to 0.46 representing "true polymorphisms", while three had MAF < or = 0.01. Eleven variants were in the promoter region; several putative transcription factor binding sites were found at these sites by database searches. Differential binding of transcription factors was demonstrated at two polymorphic sites by electrophoretic mobility shift assays, both in amniotic and HeLa cell nuclear extracts. Differential binding of the transcription factor GATA1 at -689C>G site was confirmed by a supershift. CONCLUSION: The promoter sequences of PI3 have a high degree of variability. Functional promoter variants provide a possible mechanism for explaining the differences in PI3 mRNA expression levels in the chorioamniotic membranes, and are also likely to be useful in elucidating the role of PI3 in other diseases. PMID- 16719917 TI - Cadherin2 (N-cadherin) plays an essential role in zebrafish cardiovascular development. AB - BACKGROUND: Cadherins are cell surface adhesion molecules that play important roles in development of vertebrate tissues and organs. We studied cadherin2 expression in developing zebrafish heart using in situ hybridization and immunocytochemical methods, and we found that cadherin2 was strongly expressed by the myocardium of the embryonic zebrafish. To gain insight into cadherin2 role in the formation and function of the heart, we analyzed cardiac differentiation and performance in a cadherin2 mutant, glass onion (glo). RESULTS: We found that the cadherin2 mutant had enlarged pericardial cavity, disorganized atrium and ventricle, and reduced expression of a ventricular specific marker vmhc. Individual myocardiocytes in the glo mutant embryos became round shaped and loosely aggregated. In vivo measurements of cardiac performance revealed that the mutant heart had significantly reduced heart rate, stroke volume and cardiac output compared to control embryos. Formation of the embryonic vascular system in the glo mutants was also affected. CONCLUSION: Our results suggest that cadherin2 plays an essential role in zebrafish cardiovascular development. Although the exact mechanisms remain unknown as to the formation of the enlarged pericardium and reduced peripheral blood flow, it is clear that myocardiocyte differentiation and physiological cardiovascular performance is impaired when cadherin2 function is disrupted. PMID- 16719918 TI - Effect of Ebola virus proteins GP, NP and VP35 on VP40 VLP morphology. AB - Recently we described a role for Ebola virus proteins, NP, GP, and VP35 in enhancement of VP40 VLP budding. To explore the possibility that VLP structure was altered by co-expression of EBOV proteins leading to the observed enhancement of VP40 VLP budding, we performed density gradient analysis as well as electron microscopy studies. Our data suggest that VP40 is the major determinant of VLP morphology, as co-expression of NP, GP and VP35 did not significantly change VLP density, length, and diameter. Ultra-structural changes were noted in the core of the VLPs when NP was co-expressed with VP40. Overall, these findings indicate that major changes in morphology of VP40 VLPs were likely not responsible for enhanced budding of VP40 VLPs in the presence of GP, NP and/or VP35. PMID- 16719919 TI - Study protocol: a double blind placebo controlled trial examining the effect of domperidone on the composition of breast milk [NCT00308334]. AB - BACKGROUND: Domperidone, a drug that enhances upper gastric motility, is an anti dopaminergic medication that also elevates prolactin levels. It has been shown to safely increase the milk supply of lactating women. To date, researchers have analyzed the effects of domperidone on lactating woman with respect to the quantity of their milk production, adverse effects, and drug levels in the breast milk. However, the effect of domperidone on the macronutrient composition of breast milk has not been studied and current guidelines for fortification of human milk for premature infants do not distinguish between those women using or those not using domperidone. The purpose of this study is to evaluate the effect of domperidone (given to lactating mothers of very preterm infants) on the macronutrient composition of breast milk. METHODS/DESIGN: Mothers of infants delivered at less than 31 weeks gestation, who are at least 3 weeks postpartum, and experiencing lactational failure despite non-pharmacological interventions, will be randomized to receive domperidone (10 mg three times daily) or placebo for a 14-day period. Breast milk samples will be obtained the day prior to beginning treatment and on days 4, 7 and 14. The macronutrient (protein, fat, carbohydrate and energy) and macromineral content (calcium, phosphorus and sodium) will be analyzed and compared between the two groups. Additional outcome measures will include milk volumes, serum prolactin levels (measured on days 0, 4, and 10), daily infant weights and breastfeeding rates at 2 weeks post study completion and at discharge. Forty-four participants will be recruited into the study. Analysis will be carried out using the intention to treat approach. DISCUSSION: If domperidone causes significant changes to the nutrient content of breast milk, an alteration in feeding practices for preterm infants may need to be made in order to optimize growth, nutrition and neurodevelopment outcomes. PMID- 16719920 TI - Recurrent bacteremia with Helicobacter cinaedi: case report and review of the literature. AB - BACKGROUND: Helicobacter cinaedi is a rare pathogen in humans, occurring mostly in immuno-compromised patients, with a high potential for recurrence. We describe a case of a patient with lymphoma hospitalized for chemotherapy. CASE PRESENTATION: At admission, the patient presented with an indolent and non prurigenic macular rash around her implantable venous access device. Gram staining of blood cultures revealed the presence of spiral-shaped gram-negative rods that could not be grown upon subculture. Helicobacter cinaedi was identified by PCR. No other symptoms or pathology were observed in a whole body CT scan. The implantable venous access device was removed and empiric therapy by ceftriaxone and gentamicin for 2 weeks was initiated, followed by peroral clarithromycin 2 x 500 mg/day and later by levofloxacin 2 x 500 mg/day for 7 weeks. Oncologic remission was achieved 3 months later. However, the patient was re-hospitalized 2 months later for fever, shivering, reappearance of the macular non-prurigenic rash, diarrhea, cough and asthenia. Blood cultures grew H. cinaedi. Multiple investigations could not identify the source. Empiric antibiotic therapy of ceftriaxone and doxycycline was started for 2 weeks with resolution of symptoms, followed by an oral combination of amoxicillin, metronidazole and doxycycline for 2 months; doxycycline was continued for another month. Bacteremia has not recurred for a period of 19 months. CONCLUSION: Although H. cinaedi is considered to be a low virulent bacteria, its potential to cause recurrent bacteremia should not be underestimated. H. cinaedi could have an endovascular source of infection and should be treated for an adequate duration with combined antibiotherapy. PMID- 16719921 TI - Biosynthesis of isoprenoids, polyunsaturated fatty acids and flavonoids in Saccharomyces cerevisiae. AB - Industrial biotechnology employs the controlled use of microorganisms for the production of synthetic chemicals or simple biomass that can further be used in a diverse array of applications that span the pharmaceutical, chemical and nutraceutical industries. Recent advances in metagenomics and in the incorporation of entire biosynthetic pathways into Saccharomyces cerevisiae have greatly expanded both the fitness and the repertoire of biochemicals that can be synthesized from this popular microorganism. Further, the availability of the S. cerevisiae entire genome sequence allows the application of systems biology approaches for improving its enormous biosynthetic potential. In this review, we will describe some of the efforts on using S. cerevisiae as a cell factory for the biosynthesis of high-value natural products that belong to the families of isoprenoids, flavonoids and long chain polyunsaturated fatty acids. As natural products are increasingly becoming the center of attention of the pharmaceutical and nutraceutical industries, the use of S. cerevisiae for their production is only expected to expand in the future, further allowing the biosynthesis of novel molecular structures with unique properties. PMID- 16719922 TI - Combined fluticasone propionate and salmeterol reduces RSV infection more effectively than either of them alone in allergen-sensitized mice. AB - BACKGROUND: Respiratory syncytial virus (RSV) infection is the major cause of bronchiolitis in infants and is a risk factor for the development of asthma. Allergic asthmatics are more susceptible to RSV infection and viral exacerbation. METHODS: Since the effectiveness of corticosteroids in treating RSV infection has been controversial, we tested fluticasone propionate (FP) and salmeterol (Sal) alone versus FP plus Sal (FPS) on RSV-induced airway inflammation. Mice were sensitized and challenged with ovalbumin (OVA) and infected with RSV. Following infection they were treated with FP, Sal, or FPS intranasally and airway hyperreactivity (AHR), inflammation and RSV titers were examined. RESULTS: The group treated with FPS showed significantly lower AHR compared to the group treated with FP or Sal alone. The group treated with FP alone showed slightly decreased (non-significant) AHR compared to controls. Treatment with FPS resulted in significant decreases in the percentage of eosinophils and neutrophils in bronchoalveolar lavage fluid and in lung pathology compared to FP or Sal. FP alone decreased eosinophils but not neutrophils or lymphocytes, while Sal alone decreased eosinophils and neutrophils but not lymphocytes. FPS treatment of mice infected with RSV in the absence of allergen sensitization resulted in a 50% decrease of RSV titer in the lung and a reduction in neutrophils compared to FP or Sal. CONCLUSION: Together, these results indicate that fluticasone in combination with salmeterol is a more effective treatment for decreasing airway hyperreactivity and inflammation than either of them alone in allergen sensitized, RSV-infected mice. PMID- 16719923 TI - VPg of murine norovirus binds translation initiation factors in infected cells. AB - BACKGROUND: Norovirus genomic and subgenomic RNAs are covalently linked at the 5' nucleotide to a 15 kD protein called VPg. VPg of two human norovirus strains binds translation initiation factor eIF3 and other eIFs in vitro, suggesting VPg functions in initiation of protein synthesis on viral RNA. Human norovirus strains are not cultivable, and thus experimental evidence of interactions between VPg and eIFs in infected cells has been lacking. We used the cultivable murine norovirus MNV-1 as a model to study interactions between VPg and eIFs in infected cells. RESULTS: As shown previously for human norovirus VPg, MNV-1 VPg bound eIF3, eIF4GI, eIF4E, and S6 ribosomal protein in cell extracts by GST pull down assay. Importantly, MNV-1 VPg co-precipitated eIF4GI and eIF4E from infected macrophages, providing evidence that VPg interacts with components of the translation machinery in norovirus infected cells. CONCLUSION: The interactions between MNV-1 VPg and eIFs completely mimic those reported for the human norovirus VPg, illustrating the utility of MNV-1 as a relevant molecular model to study mechanisms of human norovirus replication. PMID- 16719924 TI - Disordered semantic representation in schizophrenic temporal cortex revealed by neuromagnetic response patterns. AB - BACKGROUND: Loosening of associations and thought disruption are key features of schizophrenic psychopathology. Alterations in neural networks underlying this basic abnormality have not yet been sufficiently identified. Previously, we demonstrated that spatio-temporal clustering of magnetic brain responses to pictorial stimuli map categorical representations in temporal cortex. This result has opened the possibility to quantify associative strength within and across semantic categories in schizophrenic patients. We hypothesized that in contrast to controls, schizophrenic patients exhibit disordered representations of semantic categories. METHODS: The spatio-temporal clusters of brain magnetic activities elicited by object pictures related to super-ordinate (flowers, animals, furniture, clothes) and base-level (e.g. tulip, rose, orchid, sunflower) categories were analysed in the source space for the time epochs 170-210 and 210 450 ms following stimulus onset and were compared between 10 schizophrenic patients and 10 control subjects. RESULTS: Spatio-temporal correlations of responses elicited by base-level concepts and the difference of within vs. across super-ordinate categories were distinctly lower in patients than in controls. Additionally, in contrast to the well-defined categorical representation in control subjects, unsupervised clustering indicated poorly defined representation of semantic categories in patients. Within the patient group, distinctiveness of categorical representation in the temporal cortex was positively related to negative symptoms and tended to be inversely related to positive symptoms. CONCLUSION: Schizophrenic patients show a less organized representation of semantic categories in clusters of magnetic brain responses than healthy adults. This atypical neural network architecture may be a correlate of loosening of associations, promoting positive symptoms. PMID- 16719925 TI - Can the ubiquitous power of mobile phones be used to improve health outcomes in developing countries? AB - BACKGROUND: The ongoing policy debate about the value of communications technology in promoting development objectives is diverse. Some view computer/web/phone communications technology as insufficient to solve development problems while others view communications technology as assisting all sections of the population. This paper looks at evidence to support or refute the idea that fixed and mobile telephones is, or could be, an effective healthcare intervention in developing countries. METHODS: A Web-based and library database search was undertaken including the following databases: MEDLINE, CINAHL, (nursing & allied health), Evidence Based Medicine (EBM), POPLINE, BIOSIS, and Web of Science, AIDSearch (MEDLINE AIDS/HIV Subset, AIDSTRIALS & AIDSDRUGS) databases. RESULTS: Evidence can be found to both support and refute the proposition that fixed and mobile telephones is, or could be, an effective healthcare intervention in developing countries. It is difficult to generalize because of the different outcome measurements and the small number of controlled studies. There is almost no literature on using mobile telephones as a healthcare intervention for HIV, TB, malaria, and chronic conditions in developing countries. Clinical outcomes are rarely measured. Convincing evidence regarding the overall cost-effectiveness of mobile phone " telemedicine" is still limited and good-quality studies are rare. Evidence of the cost effectiveness of such interventions to improve adherence to medicines is also quite weak. CONCLUSION: The developed world model of personal ownership of a phone may not be appropriate to the developing world in which shared mobile telephone use is important. Sharing may be a serious drawback to use of mobile telephones as a healthcare intervention in terms of stigma and privacy, but its magnitude is unknown. One advantage, however, of telephones with respect to adherence to medicine in chronic care models is its ability to create a multi-way interaction between patient and provider(s) and thus facilitate the dynamic nature of this relationship. Regulatory reforms required for proper operation of basic and value-added telecommunications services are a priority if mobile telecommunications are to be used for healthcare initiatives. PMID- 16719926 TI - PRED(TAP): a system for prediction of peptide binding to the human transporter associated with antigen processing. AB - BACKGROUND: The transporter associated with antigen processing (TAP) is a critical component of the major histocompatibility complex (MHC) class I antigen processing and presentation pathway. TAP transports antigenic peptides into the endoplasmic reticulum where it loads them into the binding groove of MHC class I molecules. Because peptides must first be transported by TAP in order to be presented on MHC class I, TAP binding preferences should impact significantly on T-cell epitope selection. DESCRIPTION: PRED(TAP) is a computational system that predicts peptide binding to human TAP. It uses artificial neural networks and hidden Markov models as predictive engines. Extensive testing was performed to valid the prediction models. The results showed that PRED(TAP) was both sensitive and specific and had good predictive ability (area under the receiver operating characteristic curve Aroc>0.85). CONCLUSION: PRED(TAP) can be integrated with prediction systems for MHC class I binding peptides for improved performance of in silico prediction of T-cell epitopes. PRED(TAP) is available for public use at [1]. PMID- 16719927 TI - Causal inference of regulator-target pairs by gene mapping of expression phenotypes. AB - BACKGROUND: Correlations between polymorphic markers and observed phenotypes provide the basis for mapping traits in quantitative genetics. When the phenotype is gene expression, then loci involved in regulatory control can theoretically be implicated. Recent efforts to construct gene regulatory networks from genotype and gene expression data have shown that biologically relevant networks can be achieved from an integrative approach. In this paper, we consider the problem of identifying individual pairs of genes in a direct or indirect, causal, trans acting relationship. RESULTS: Inspired by epistatic models of multi-locus quantitative trait (QTL) mapping, we propose a unified model of expression and genotype to identify quantitative trait genes (QTG) by extending the conventional linear model to include both genotype and expression of regulator genes and their interactions. The model provides mapping of specific genes in contrast to standard linkage approaches that implicate large QTL intervals typically containing tens of genes. In simulations, we found that the method can often detect weak trans-acting regulators amid the background noise of thousands of traits and is robust to transcription models containing multiple regulator genes. We reanalyze several pleiotropic loci derived from a large set of yeast matings and identify a likely alternative regulator not previously published. However, we also found that many regulators can not be so easily mapped due to the presence of cis-acting QTLs on the regulators, which induce close linkage among small neighborhoods of genes. QTG mapped regulator-target pairs linked to ARN1 were combined to form a regulatory module, which we observed to be highly enriched in iron homeostasis related genes and contained several causally directed links that had not been identified in other automatic reconstructions of that regulatory module. Finally, we also confirm the surprising, previously published results that regulators controlling gene expression are not enriched for transcription factors, but we do show that our more precise mapping model reveals functional enrichment for several other biological processes related to the regulation of the cell. CONCLUSION: By incorporating interacting expression and genotype, our QTG mapping method can identify specific regulator genes in contrast to standard QTL interval mapping. We have shown that the method can recover biologically significant regulator-target pairs and the approach leads to a general framework for inducing a regulatory module network topology of directed and undirected edges that can be used to identify leads in pathway analysis. PMID- 16719928 TI - Posttraumatic stress disorder among female street-based sex workers in the greater Sydney area, Australia. AB - BACKGROUND: This paper examines rates of exposure to work-related violence and other trauma, and the prevalence of lifetime and current posttraumatic stress disorder (PTSD) among female street-based sex workers. It also investigates associations between current PTSD symptoms and: demographic characteristics, psychiatric comorbidity, injecting and sex risk behaviours, and trauma history. METHODS: Cross sectional data collected from 72 women via face to face structured interviews. The interview included structured diagnostic assessment of DSM-IV PTSD; drug dependence; depression; experience of childhood trauma; and an assessment of sex working history. RESULTS: All but one of the women interviewed reported experiencing trauma, with the majority reporting multiple traumas that typically began in early childhood. Child sexual abuse, adult sexual assault and work related violence were commonly reported. Just under half of the women met DSM-IV criteria for PTSD and approximately one-third reported current PTSD symptoms. Adult sexual assault was associated with current PTSD symptoms. Depression and drug dependence were also highly prevalent; cocaine dependence in particular was associated with elevated rates of injecting risk and sexual risk behaviours. CONCLUSION: These women reported complex trauma histories and despite ongoing opportunities for clinical intervention, they continued to experience problems, suggesting that current models of treatment may not be appropriate. More targeted interventions, and integrated mental health and drug treatment services are needed to address the problems these women are experiencing. Outreach services to these women remain a priority. Education strategies to reduce risky injecting and sexual behaviours among sex workers should also remain a priority. PMID- 16719929 TI - Induction of revertant fibres in the mdx mouse using antisense oligonucleotides. AB - BACKGROUND: Duchenne muscular dystrophy is a fatal genetic disorder caused by dystrophin gene mutations that result in premature termination of translation and the absence of functional protein. Despite the primary dystrophin gene lesion, immunostaining studies have shown that at least 50% of DMD patients, mdx mice and a canine model of DMD have rare dystrophin-positive or 'revertant' fibres. Fine epitope mapping has shown that the majority of transcripts responsible for revertant fibres exclude multiple exons, one of which includes the dystrophin mutation. METHODS: The mdx mouse model of muscular dystrophy has a nonsense mutation in exon 23 of the dystrophin gene. We have shown that antisense oligonucleotides (AOs) can induce the removal of this exon, resulting in an in frame mRNA transcript encoding a shortened but functional dystrophin protein. To emulate one exonic combination associated with revertant fibres, we target multiple exons for removal by the application of a group of AOs combined as a "cocktail". RESULTS: Exons 19-25 were consistently excluded from the dystrophin gene transcript using a cocktail of AOs. This corresponds to an alternatively processed gene transcript that has been sporadically detected in untreated dystrophic mouse muscle, and is presumed to give rise to a revertant dystrophin isoform. The transcript and the resultant correctly localised smaller protein were confirmed by RT-PCR, immunohistochemistry and western blot analysis. CONCLUSION: This work demonstrates the feasibility of AO cocktails to by-pass dystrophin mutation hotspots through multi-exon skipping. Multi-exon skipping could be important in expediting an exon skipping therapy to treat DMD, so that the same AO formulations may be applied to several different mutations within particular domains of the dystrophin gene. PMID- 16719930 TI - An exploratory study of GP awareness of carer emotional needs in Western Australia. AB - BACKGROUND: The impact of caring for a family member or friend is a life changing experience. Often carers are struggling to cope with ongoing demands of caring for someone. At some point, most carers will approach their family physician for advice on aspects of their role. Carers Western Australia sought information and opinions of General Practitioners (GPs) regarding being a carer. This exploratory survey was to assess the perceptions of Western Australian GPs regarding their role in providing information and support to carers and their awareness of carer needs and issues. METHODS: A telephone survey design of an opportunistic sample of 66 Western Australian GPs was conducted. The responses were both closed and open-ended questions to allow for probing of responses. Responses were analysed using both qualitative and quantitative analysis. RESULTS: GPs are generally aware of their role in regard to carers and most doctors (88%) said that they had been approached for help in accessing services by carers. A majority of respondents said that carers and spoken to them (70%) and asked for help with (77%) emotional needs. However, when asked how these needs are met, GPs tended to provide practical assistance for the care recipient and carer as a means of addressing those needs. This primarily included providing referral to services to ensure that the carer has practical assistance in caring for the person. However, GPs are less able to provide the necessary emotional or psychological support needed by carers before crisis point is reached. Most doctors said that they had experienced difficulties in providing assistance to carers (84%) at some time. CONCLUSION: GPs may be unable or unwilling to provide the necessary assistance to carers who are showing signs of carer burnout and stress. The GP needs to adopt a more holistic approach when treating a patient as to the interaction with the caregiver. PMID- 16719931 TI - A pilot trial on safety and efficacy of erythrocyte-mediated steroid treatment in CF patients. AB - BACKGROUND: Chronic neutrophil inflammation of the respiratory tract tissues plays a key role in the pathogenesis and in prognosis of cystic fibrosis (CF). It is evident that an anti-inflammatory therapy represents an important step in the treatment of CF patients. Corticosteroids and ibuprofen have been proven to slow down the impairment of the pulmonary function in CF patients but their use is limited by the frequency of adverse events. A novel strategy for delivering low doses of steroids for long periods through the infusion of autologous erythrocytes loaded with dexamethasone has been recently set up. A recent study suggested the feasibility of therapy with low doses of corticosteroids delivered through engineered erythrocytes in CF patients. This study presents a further analysis of safety and efficacy of this therapy. METHODS: The treatment group was not randomised and the assignment was based on the patient's consent. Patients entered the study if they had a forced expiratory volume in 1 second (FEV1) <70%, puberty development completed, pancreatic insufficiency, and chronic pulmonary infection requiring frequent cycles of intravenous antibiotic therapy. Patients were excluded if they underwent systemic corticosteriod therapy in the three months preceding the experimental treatment or were on therapy with non-steroidal anti inflammatory drugs (NASDs), or if they had liver CF disease, allergic bronchopulmonary aspergillosis, or positive tuberculin test. Controls were patients who followed a standard treatment, who fulfilled the enrollment criteria, and who were matched to the experimental group by gender, age, and severity of the disease. RESULTS: Nine patients in the experimental group received the treatment once a month for a period of 24 month. Patients did not develop diabetes, cataract, or hypertension, or other typical side effects of steroid treatment during the follow up period. There was a constant improvement of FEV1 in patients undergoing the experimental treatment compared to a gradual decrease of the same parameter in the standard therapy group (P = 0.04). The average of clinic and radiological indexes did not vary. The number of infective relapses that have required antibiotic intravenous therapy was not different in the two groups, although the average of these episodes was slightly higher in the experimental therapy group. CONCLUSION: Intraerythrocyte corticosteroid treatment may stabilize the respiratory function in CF patients but is often considered too invasive by patients. The results obtained by our study may help planning an experimental, controlled, randomised study. A sample size of 150 patients per group would be sufficient for demonstrating such a difference with a 95% confidence interval and a power of 90%. PMID- 16719932 TI - Intron gain and loss in segmentally duplicated genes in rice. AB - BACKGROUND: Introns are under less selection pressure than exons, and consequently, intronic sequences have a higher rate of gain and loss than exons. In a number of plant species, a large portion of the genome has been segmentally duplicated, giving rise to a large set of duplicated genes. The recent completion of the rice genome in which segmental duplication has been documented has allowed us to investigate intron evolution within rice, a diploid monocotyledonous species. RESULTS: Analysis of segmental duplication in rice revealed that 159 Mb of the 371 Mb genome and 21,570 of the 43,719 non-transposable element-related genes were contained within a duplicated region. In these duplicated regions, 3,101 collinear paired genes were present. Using this set of segmentally duplicated genes, we investigated intron evolution from full-length cDNA supported non-transposable element-related gene models of rice. Using gene pairs that have an ortholog in the dicotyledonous model species Arabidopsis thaliana, we identified more intron loss (49 introns within 35 gene pairs) than intron gain (5 introns within 5 gene pairs) following segmental duplication. We were unable to demonstrate preferential intron loss at the 3' end of genes as previously reported in mammalian genomes. However, we did find that the four nucleotides of exons that flank lost introns had less frequently used 4-mers. CONCLUSION: We observed that intron evolution within rice following segmental duplication is largely dominated by intron loss. In two of the five cases of intron gain within segmentally duplicated genes, the gained sequences were similar to transposable elements. PMID- 16719935 TI - The struggle to detect circulating DNA. AB - In various diseases, such as cancer, autoimmune disease, sepsis or myocardial infarction, elevated levels of circulating DNA can be measured. However, its predictive value is under debate. Circulating DNA in plasma is protein-bound (nucleosomal) DNA. Quantification of circulating DNA can be performed by real time quantitative PCR or immunological methods such as ELISA. The diagnostic value of both methods can be impaired by inappropriate handling of the samples. Assessment of circulating DNA in patients admitted to the intensive care unit offers a tool for predicting morbidity and mortality. PMID- 16719933 TI - Expression of bioactive bone morphogenetic proteins in the subacromial bursa of patients with chronic degeneration of the rotator cuff. AB - Degeneration of the rotator cuff is often associated with inflammation of the subacromial bursa and focal mineralization of the supraspinatus tendon. Portions of the supraspinatus tendon distant from the insertion site could transform into fibrous cartilage, causing rotator-cuff tears owing to mechanical instability. Indirect evidence is presented to link this pathology to ectopic production and secretion of bioactive bone morphogenetic proteins (BMPs) from sites within the subacromial bursa. Surgically removed specimens of subacromial bursa tissue from patients with chronic tears of the rotator cuff were analyzed by immunohistochemistry and reverse transcription-PCR. Bioactive BMP was detected in bursa extracts by a bioassay based on induction of alkaline phosphatase in the osteogenic/myogenic cell line C2C12. Topical and differential expression of BMP 2/4 and BMP-7 mRNA and protein was found in bursa tissue. The bioassay of C2C12 cells revealed amounts of active BMP high enough to induce osteogenic cell types, and blocking BMP with specific antibodies or soluble BMP receptors Alk-3 and Alk 6 abolished the inductive properties of the extract. Sufficient information was gathered to explain how ectopic expression of BMP might induce tissue transformation into ectopic bone/cartilage and, therefore, promote structural degeneration of the rotator cuff. Early surgical removal of the subacromial bursa might present an option to interrupt disease progression. PMID- 16719937 TI - Systems biology of gene regulation fulfills its promise. PMID- 16719936 TI - Aspects of early arthritis. Traditional DMARD therapy: is it sufficient? AB - There is increasing evidence for beneficial effects of early DMARD (disease modifying antirheumatic drug) therapy over delayed treatment in patients who present with arthritis of recent onset. However, no universal consensus exists concerning the choice of initial drug or whether single drugs or combinations should be given as initial treatments. Recent studies have focused on the benefits of various strategies in which treatments were tailored to achieve low levels of disease activity, as assessed using validated response criteria. These studies demonstrated superiority of 'aggressive' over 'conventional' approaches. Whether the inclusion of tumour necrosis factor antagonists or other biologic targeted therapies in such strategies confers additional benefits in terms of improved long-term outcomes must be clarified by further studies. Assessment of risks in the individual patient, allowing individual 'tailoring' of the initial treatment, would be desirable. PMID- 16719938 TI - Stem cells in the genomic age. PMID- 16719940 TI - The obstetric forceps: shrouded in mystery. PMID- 16719941 TI - A conceptual model for the pathophysiology of vulvar vestibulitis syndrome. AB - Vulvar vestibulitis syndrome (vestibulitis), the most common type of chronic vulvovaginal pain, impairs the psychologic, physical, and reproductive health of approximately 10% of women at some point in their lives. Research on the pathophysiology of vestibulitis suggests abnormalities in 3 interdependent systems: vestibular mucosa, pelvic floor muscles, and central nervous system pain regulatory pathways. To date, causes and relative contributions of these abnormalities to the development and maintenance of vestibulitis remain poorly understood. Research consistently supports the conceptualization of vestibulitis as a chronic pain disorder-akin to fibromyalgia, irritable bowel disorder, and temporomandibular disorder (TMD)-that is far more complex than vestibular hypersensitivity alone. Nevertheless, the clinical diagnosis of vestibulitis continues to rely on subjective report of pain during intercourse and vestibular sensitivity on clinical examination after exclusion of other gynecologic disorders. We propose that current diagnostic criteria, which are based on highly subjective patient and clinician measures, are not sufficient to describe and properly classify the heterogeneous clinical presentations of this disorder. To inform clinical care or research, we must be able to objectively characterize women with vestibulitis. This narrative review critically appraises current conceptualization of vestibulitis and presents a context for studying vestibulitis as a chronic pain disorder, emphasizing the need for objective assessment of clinical features. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader should be able to state that vulvar vestibulitis is common; recall that the disorder has three major pathophysiological pathways and that understanding of these pathways is important in selecting treatment options, and explain that the clinician must attempt to properly classify the clinical presentations of the disorder. PMID- 16719943 TI - Subclinical hypothyroidism and pregnancy. AB - Pregnancy has a considerable impact on thyroid homeostasis that complicates the diagnosis of hypothyroidism. Pregnant women with overt hypothyroidism have appreciable maternal morbidity as well as perinatal morbidity and mortality. Treatment of such women has been shown to improve these outcomes. Controversy regarding the importance of identifying and treating women with subclinical hypothyroidism erupted after several reports have linked variously defined hypothyroidism with impaired neurodevelopment of the fetus. Frequently, these reports are erroneously considered as identifying women with subclinical hypothyroidism, when in fact, none of these studies specifically identified such women. To date, there have been no well-controlled, long-term studies on offspring of pregnant women with subclinical hypothyroidism as contemporaneously defined. Furthermore, these reports have led to conflicting and confusing position statements from several national entities as to whether all pregnant women should be screened for hypothyroidism and treatment prescribed for those found to have subclinical hypothyroidism. Importantly, there are also no published intervention trials specifically assessing the efficacy of such treatment to improve neurodevelopmental outcomes in offspring of women with subclinical hypothyroidism. It is acknowledged that obstetricians are under increasing pressure to screen for and treat pregnant women with subclinical hypothyroidism despite uncertainty that their offspring would benefit from therapy. National endocrine groups recommending such screening and treatment have emphasized that there is a need for large clinical trials to address these issues. Until such studies are completed, routine screening for and treatment of subclinical hypothyroidism during pregnancy is unwarranted and should be considered experimental. TARGET AUDIENCE: Obstetricians and Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader should be able to recall the difficulty in identifying and treating women with subclinical hypothyroidism, summarize the current thinking about the laboratory diagnosis of subclinical hypothyroidism, explain that there are no well controlled studies that indicate that treatment during pregnancy will improve neurodevelopmental outcome, and state that screening during pregnancy at this time is unwarranted. PMID- 16719939 TI - Clinical review: thyroid hormone replacement in children after cardiac surgery- is it worth a try? AB - Cardiac surgery using cardiopulmonary bypass produces a generalized systemic inflammatory response, resulting in increased postoperative morbidity and mortality. Under these circumstances, a typical pattern of thyroid abnormalities is seen in the absence of primary disease, defined as sick euthyroid syndrome (SES). The presence of postoperative SES mainly in small children and neonates exposed to long bypass times and the pharmacological profile of thyroid hormones and their effects on the cardiovascular physiology make supplementation therapy an attractive treatment option to improve postoperative morbidity and mortality. Many studies have been performed with conflicting results. In this article, we review the important literature on the development of SES in paediatric postoperative cardiac patients, analyse the existing information on thyroid hormone replacement therapy in this patient group and try to summarize the findings for a recommendation. PMID- 16719942 TI - Intrauterine insemination versus timed intercourse for cervical hostility in subfertile couples. AB - The postcoital test has poor diagnostic and prognostic characteristics. Nevertheless, some physicians believe it can identify scanty or abnormal mucus that might impair fertility. One way to avoid "hostile" cervical mucus is intrauterine insemination. With this technique, the physician injects sperm directly into the uterine cavity through a small catheter passed through the cervix; the theory is to bypass the "hostile" cervical mucus. Although most gynecologic societies do not endorse use of intrauterine insemination for hostile cervical mucus, some physicians consider it an effective treatment for women with infertility thought the result of cervical mucus problems. The aim of this review was to determine the effectiveness of intrauterine insemination with or without ovarian stimulation in women with cervical hostility who failed to conceive.We searched Cochrane Central Register of Controlled Trials (CENTRAL) on The Cochrane Library Issue 2, 2005, MEDLINE (1966 to June 2005), EMBASE (1980 to June 2005), POPLINE (to June 2005), and LILACS (to June 2005). In addition, we contacted experts and searched the reference list of relevant articles and book chapters. We included randomized and quasirandomized, controlled trials comparing intrauterine insemination with intercourse timed at the presumed fertile period. Participants were women with cervical hostility who failed to conceive for at least 1 year. We assessed the titles and abstracts of 386 publications and 2 reviewers independently abstracted data on methods and results from 5 studies identified for inclusion. The main outcome is pregnancy rate per couple. We did not pool the outcomes of the included 5 studies in a meta-analysis resulting from the methodological quality of the trials and variations in the patient characteristics and interventions. Narrative summaries of the outcomes are provided. Each study was too small for a clinically relevant conclusion. None of the studies provided information on important outcomes such as spontaneous abortion, multiple pregnancies, and ovarian hyperstimulation syndrome. There is no evidence from the published studies that intrauterine insemination is an effective treatment for cervical hostility. Given the poor diagnostic and prognostic properties of the postcoital test and the observation that the test has no benefit on pregnancy rates, intrauterine insemination (with or without ovarian stimulation) is unlikely to be a useful treatment for putative problems identified by postcoital testing. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader should be able to recall that there is a lack of adequate studies that support that intrauterine insemination (IUI) is an effective treatment of cervical hostility, explain that the postcoital test has poor diagnostic and prognostic properties, and state that the use of both tests has no benefit on pregnancy rates. EDITOR'S NOTE: Although many assisted reproductive technology (ART) programs no longer perform postcoital tests, many perform intrauterine insemination (IUI), often with gonadotropins or clomiphene citrate, in their subfertile patients. Therefore, this review article will be of value to our readers who treat subfertile patients with IUI, whether or not they perform postcoital tests. For additional explanations of the statistical tests employed in this review, see D. Grimes, KF Schulz, Obstetrical and Gynecologic Survey, 57; Supplement 3: S35, September 2002; and D. Grimes, KF Schulz, Obstetrical and Gynecologic Survey, Supplement 2, S53-S69, September 2005.-RBJ. PMID- 16719944 TI - Developmental capacity of Antarctic minke whale ( Balaenoptera bonaerensis ) vitrified oocytes following in vitro maturation, and parthenogenetic activation or intracytoplasmic sperm injection. AB - The present study investigated the effects of the sexual maturity of oocyte donors on in vitro maturation (IVM) and the parthenogenetic developmental capacity of fresh minke whale oocytes. The effects of cytochalasin B (CB) pretreatment and two types of cryoprotectant solutions (ethylene glycol (EG) or ethylene glycol and dimethylsulfoxide (EG + DMSO)) on the in vitro maturation of vitrified immature whale oocytes were compared, and the developmental capacity of vitrified immature whale oocytes following IVM and intracytoplasmic sperm injection examined (ICSI). The maturation rate did not differ significantly with sexual maturity (adult, 60.9%; prepubertal, 53.1%), but the parthenogenetic activation rate of oocytes from adult donors (76.7%) was significantly higher (p < 0.05) than that of oocytes from prepubertal donors (46.4%). The maturation rates after vitrification and warming were not significantly different between the EG (22.2%) and EG + DMSO groups (30.2%), or between the CB-treated (30.4%) and non-CB-treated groups (27.3%). These results indicate that parthenogenetic activation of in vitro matured oocytes from adult minke whales was superior to that from prepubertal whales, but that the developmental capacity of the whale oocytes after parthenogenetic activation or ICSI was still low. The present study also showed that CB treatment before vitrification and two kinds of cryoprotectants did not improve the IVM rate following the vitrification of immature whale oocytes. PMID- 16719945 TI - Behaviour of the vitelline envelope in Bufo arenarum oocytes matured in vitro in blockade to polyspermy. AB - During activation of amphibian eggs, cortical granule exocytosis causes elaborate ultrastructural changes in the vitelline envelope. These changes involve modifications in the structure of the vitelline envelope and formation of a fertilization envelope (FE) that can no longer be penetrated by sperm. In Bufo arenarum, as the egg traverses the oviduct, the vitelline envelope is altered by a trypsin-like protease secreted by the oviduct, which induces an increased susceptibility of the vitelline envelope to sperm lysins. Full-grown oocytes of B. arenarum, matured in vitro by progesterone, are polyspermic, although cortical granule exocytosis seems to occur within a normal chronological sequence. These oocytes can be fertilized with or without trypsin treatment, suggesting that the vitelline envelope is totally sperm-permeable. Vitelline envelopes without trypsin treatment cannot retain either gp90 or gp96. This suggests that these glycoproteins are involved in the block to polyspermy and that trypsin treatment of matured in vitro oocytes before insemination is necessary to enable vitelline envelopes to block polyspermy. The loss of the binding capacity in vitelline envelopes isolated from B. arenarum oocytes matured in vitro with trypsin treatment and activated by electric shock suggests that previous trypsin treatment is a necessary step for sperm block to occur. When in vitro matured oocytes were incubated with the product of cortical granules obtained from in vitro matured oocytes (vCGP), vitelline envelopes with trypsin treatment were able to block sperm entry. These oocytes exhibited the characteristic signs of activation. These results support the idea that B. arenarum oocytes can be activated by external stimuli and suggest the presence of unknown oocyte surface receptors linked to the activation machinery in response to fertilization. Electrophoretic profiles obtained by SDS-PAGE of solubilized vitelline envelopes from oocytes matured in vitro revealed the conversion of gp40 (in vitro matured oocytes, without trypsin treatment) to gp38 (ascribable to trypsin activity or cortical granule product activity, CGP) and the conversion of gp70 to gp68 (ascribable to trypsin activity plus CGP activity). Taking into account that only the vitelline envelopes of in vitro matured oocytes with trypsin treatment and activated can block sperm entry, we may suggest that the conversion of gp70 to gp68 is related to the changes associated with sperm binding. PMID- 16719946 TI - Expression of mRNA and protein localization of epidermal growth factor and its receptor in goat ovaries. AB - To examine the possibility that epidermal growth factor (EGF) and its receptor (EGF-R) are expressed throughout folliculogenesis, we studied the presence and distribution of EGF and EGF-R in goat ovaries. Ovaries of goats were collected and either fixed in paraformaldehyde for immunohistochemical localization of proteins, or used for the isolation of follicles, luteal cells and ovarian surface epithelium to study mRNA expression for EGF and EGF-R, using the reverse transcriptase polymerase chain reaction. EGF protein and mRNA were found in primordial, primary and secondary follicles as well as in small and large antral follicles and in surface epithelium, but in corpora lutea only the protein could be detected. Antral follicles expressed EGF mRNA in oocyte, cumulus, mural granulosa and theca cells. For EGF-R, both protein and mRNA were present at all stages of follicular development and in all antral follicular compartments. EGF-R protein and mRNA were also found in corpora lutea and surface epithelium. It is concluded that EGF and its receptor are expressed in goat ovarian follicles at all stages of follicle development, in corpora lutea, and in ovarian surface epithelium. PMID- 16719947 TI - CD9 protein appears on growing mouse oocytes at the time when they develop the ability to fuse with spermatozoa. AB - CD9 is a member of the tetraspanin superfamily proteins and is the only protein on the mouse oocyte which is known to be indispensable in sperm-egg fusion. Here, using indirect immunofluorescence we show that CD9 appears on the oolemma during the early stages of the growth of the oocyte, when it measures 13-22 microm in diameter. When the oocyte reaches a diameter of 17-22 microm, the density of CD9 in its oolemma is similar to the density of this protein in the cell membrane of the fully grown secondary oocyte. The appearance of CD9 in growing oocytes correlates with the previously reported time of the acquisition of fusibility between the spermatozoon and the egg. Accordingly we propose that during oogenesis the development of the ability of the oolemma to fuse with sperm may be regulated by synthesis of CD9 by the oocyte. PMID- 16719948 TI - Increase in DNA fragmentation and apoptosis-related gene expression in frozen thawed bovine blastocysts. AB - Evaluation of apoptosis and expression level of apoptosis-related genes is useful for examining the variation in embryo quality according to environmental change. The objective of this study was to investigate DNA fragmentation and apoptosis related gene expression patterns in frozen-thawed bovine blastocysts. In vitro produced day 7 blastocysts were frozen by two different vitrification methods (conventional 0.25 ml straw or MVC straw). After thawing, DNA fragmentation of surviving embryos was examined by TUNEL assay, and the expression patterns of their apoptotic genes (survivin, Fas, Hsp 70 and caspase-3) were evaluated using real-time quantitative reverse transcriptase polymerase chain reaction. In vitro survival rates of frozen-thawed embryos were higher following the MVC vitrification method (88.2% re-expanded at 24 h, 77.1% hatching at 48 h) than the conventional (C) vitrification method (77.0% re-expanded at 24 h, 66.7% hatching at 48 h). However, both vitrified methods resulted in a significantly higher apoptotic index (C vitrification method 11.9%, MVC vitrification method 11.0%) than in non-frozen embryos (3.0%). Expression levels of survivin, Fas, caspase-3, and Hsp 70 were also increased in the frozen-thawed embryos compared with non frozen embryos. These results indicate that the cryopreservation procedure might cause damage that results in an increase in DNA fragmentation and apoptosis related gene transcription, reducing developmental capacity of frozen-thawed embryos. PMID- 16719949 TI - Na+ /Ca2+ exchanger contributes to asterosap-induced elevation of intracellular Ca2+ concentration in starfish spermatozoa. AB - Asterosap, a group of equally active isoforms of sperm-activating peptides from the egg jelly of the starfish Asterias amurensis, functions as a chemotactic factor for sperm. It transiently increases the intracellular cGMP level of sperm, which in turn induces a transient elevation of intracellular Ca(2+) concentration ([Ca(2+)](i)). Using a fluorescent Ca(2+)-sensitive dye, Fluo-4 AM, we measured the changes in sperm [Ca(2+)](i) in response to asterosap. KB-R7943 (KB), a selective inhibitor of Na(+)/Ca(2+) exchanger (NCX), significantly inhibited the asterosap-induced transient elevation of [Ca(2+)](i), suggesting that asterosap influences [Ca(2+)](i) through activation of a K+-dependent NCX (NCKX). An NCKX activity of starfish sperm also shows K(+) dependency like other NCKXs. Therefore, we cloned an NCKX from the starfish testes and predicted that it codes for a 616 amino acid protein that is a member of the NCKX family. Pharmacological evidence suggests that this exchanger participates in the asterosap-induced Ca(2+) entry into sperm. PMID- 16719950 TI - Requirement for a localized, IP3R-generated Ca2+ transient during the furrow positioning process in zebrafish zygotes. AB - We report that the first localized Ca(2+) transient visualized in the blastodisc cortex of post-mitotic zebrafish zygotes has unique features. We confirm that this initial 'furrow positioning' Ca(2+) transient precedes the physical appearance of the first cleavage furrow at the blastodisc surface and that it has unique dynamics, which distinguish it from the subsequent furrow propagation transients that develop from it. This initial transient displays a distinct rising phase that peaks prior to the initiation of the two linear, subsurface, self-propagating Ca(2+) waves that constitute the subsequent furrow propagation transient. Through the carefully timed introduction of the Ca(2+) buffer, dibromo BAPTA, we also demonstrate the absolute requirement of this initial rising phase Ca(2+) transient in positioning the furrow at the blastodisc surface: no rising phase transient, no cleavage furrow. Likewise, the introduction of the inositol 1,4,5-trisphosphate receptor (IP3R) antagonist, 2-aminoethoxydiphenyl borate, eliminates both the rising phase transient and the appearance of the furrow at the cell surface. On the other hand, antagonists of the ryanodine receptor and NAADP-sensitive channels, or simply bathing the zygote in Ca(2+)-free medium, have no effect on the generation of the rising phase positioning transient or the appearance of the furrow at the surface. This suggests that like the subsequent propagation and deepening/zipping Ca(2+) transients, the rising phase furrow positioning transient is also generated specifically by Ca(2+) released via IP3Rs. We propose, however, that despite being generated by a similar Ca(2+) release mechanism, the unique features of this initial transient suggest that it might be a distinct signal with a specific function associated with positioning the cleavage furrow at the blastodisc surface. PMID- 16719951 TI - The localization of LAP2 beta during pronuclear formation in bovine oocytes after fertilization or activation. AB - We have shown that the assembly of lamin-associated polypeptide (LAP) 2beta was detected surrounding the chromatin mass around the time of extrusion of the second polar body (PB) in some fertilized oocytes, but not in most activated oocytes, by using A23187 and cycloheximide (CaA + CH). Here, we immunohistologically analysed the correlation between LAP2beta assembly and chromatin condensation in fertilized and activated oocytes during the second meiosis. In bovine cumulus cells, the onset of LAP2beta assembly was observed around anaphase chromosomes with strongly phosphorylated histone H3. No LAP2beta assembled around the chromosomes in the first and second polar bodies and the alternative oocyte chromatin (oCh) if histone H3 was phosphorylated. Only histone H3 of oCh was completely dephosphorylated during the telophase II/G1 transition (Tel II/G1), and then LAP2beta assembled around only the oCh without phosphorylated histone H3. In the oocytes activated by CaA + CH, LAP2beta did not assemble around the condensed oCh during the Tel II/G1 transition, although their histone H3 dephosphorylation occurred rather rapidly compared with that of the fertilized oocytes. The patterns of histone H3 dephosphorylation and LAP2beta assembly in oocytes activated by CaA alone showed greater similarity to those in fertilized oocytes than to those in oocytes activated by CaA + CH. These results show that LAP2beta assembles around only oCh after complete dephosphorylation of histone H3 after fertilization and activation using CaA alone, and that the timing of histone H3 dephosphorylation and LAP2beta assembly in these oocytes is different from that of somatic cells. The results also indicate that CH treatment inhibits LAP2beta assembly around oCh but not histone H3 dephosphorylation. PMID- 16719952 TI - Analysis of specific factors generating 2-cell block in AKR mouse embryos. AB - The phenomenon of the developmental arrest at the 2-cell stage of 1-cell embryos from some mouse strains during in vitro culture is known as the 2-cell block. We investigated the specific factors involved in the 2-cell block of AKR embryos by means of a modified culture system, the production of reconstructed embryos by pronuclear exchange and a cross experiment. In a culture medium with phosphate, 94.6% of 1-cell embryos from the C57BL mouse strain developed to the blastocyst stage, but 95.7% of embryos from the AKR mouse strain showed 2-cell block. Phosphate-free culture medium rescued the 2-cell block of AKR embryos and accelerated the first cell cycle of the embryos. Co-culture with BRL cells and a BRL-conditioned medium fractionated below 30 kDa also rescued the 2-cell block of AKR embryos. Examinations of in vitro development of reconstructed embryos and of embryos from F1 females between AKR and C57BL strains clearly demonstrated that the AKR cytoplast caused the 2-cell block. In the backcrossed female progeny between (AKR x C57BL) F1 males and AKR females, about three-quarters of the embryos were of the 2-cell blocking phenotype and about one-quarter were of the non-blocking phenotype. These results suggest that two genes are responsible for the 2-cell block of AKR embryos. PMID- 16719953 TI - Electrosurgical endoscopic cordectomy with microdissection electrodes: a comparative study with CO2 laser. AB - Microdissection electrodes (MEs) have previously been used to perform endoscopic cordectomies. We designed a prospective study in order to compare the ME with the CO2 laser technique. Over two years, 20 patients with T1 glottic carcinoma were operated on with CO2 laser and 20 with MEs. The device was chosen alternatively for each new patient. Two patients in both treatment groups had slight glottic incompetence. Three patients in each group showed web formation. The only granuloma was observed in a CO2 laser patient. Seven of the ME patients developed slight dysphonia, 10 developed medium grade dysphonia and three developed severe dysphonia. Seven of the CO2 laser patients developed slight dysphonia, seven developed medium grade dysphonia and six developed severe dysphonia. No statistically significant differences were observed on comparing the grade of dysphonia with patient age, T stage, type of cordectomy or surgical device. We consider the ME to be a useful and inexpensive alternative to CO2 laser. PMID- 16719954 TI - Auditory neuropathy in a patient with mitochondrial myopathy and multiple mtDNA deletions. AB - Auditory neuropathy (AN) is a hearing disorder characterized by the absence or severe distortion of the auditory brainstem responses, in the presence of preserved otoacoustic emissions. This peculiar combination suggests the presence of a defect impinging upon the functional complex formed by inner hair cells, the primary afferents (spiral ganglion neurones) and the first order synapses between hair cells and the cochlear nerve. Typically, AN patients show a severe speech perception impairment, which appears reduced out of proportion to pure tone threshold, but the clinical presentation of AN is quite complex. Hearing loss is a common symptom associated with mitochondrial diseases; however, AN has only rarely been reported in these disorders. Here we report a rare association, the first case observed in Italy, in a patient with autosomal recessive mitochondrial myopathy and mitochondrial DNA multiple deletions, and a hearing deficit with the audiological and electrophysiological features of AN. PMID- 16719955 TI - Squamous cell carcinoma in situ of the external auditory canal. AB - We report two cases of carcinoma in situ in the external auditory canal (EAC), presenting with symptoms such as pain, long-term itching of the ear, easy contact bleeding, canal otorrhea and hearing loss. Otoscopic examination revealed granulation tissue and a greyish-black tumour with irregular surface. The first patient had previously been diagnosed with otitis externa with persistent ear itching for the past three years. The second patient had received tympanoplasty for treatment of chronic otitis media on the right ear ten years ago. The first case was treated with wide excision, whereas the second patient received resection of the skin of the EAC together with its adjacent soft tissue, followed by skin grafting. No tumour recurrence was noted in the fourth and third post operative year for the first and second patient respectively. We suggest that EAC carcinoma can be detected early and treated. PMID- 16719956 TI - Adenoidectomy: does it work? AB - Adenoidectomy is a common, routine paediatric operation for which the evidence base for effectiveness is lacking. While there is a broad evidence base of variable quality for other common children's ENT operations, most published data including adenoidectomy is combined with the effect of combined tonsillectomy or grommet surgery. For the common indications for adenoidectomy, does it work? PMID- 16719957 TI - Distribution of Pholeter gastrophilus (Digenea) within the stomach of four odontocete species: the role of the diet and digestive physiology of hosts. AB - We compared the distribution of the digenean Pholeter gastrophilus in the stomach of 27 harbour porpoises, Phocoena phocoena, 27 striped dolphins, Stenella coeruleoalba, 18 bottlenose dolphins, Tursiops truncatus, and 100 long-finned pilot whales, Globicephala melas. The stomach of these species is composed of 4 chambers of different size, structure and function. In all species, P. gastrophilus was largely restricted to the glandular region of the stomach, but the parasite tended to favour the fundic chamber in bottlenose dolphins and harbour porpoises, the pyloric chamber in pilot whales, and none in striped dolphins. However, predictability at infrapopulation level was generally low, suggesting a weak preference of P. gastrophilus for any of the chambers. Three hypotheses were tested to investigate a common cause for the distribution of P. gastrophilus in all host species, namely, colonization of chambers was (1) sequential, (2) dependent on chamber size, or (3) dependent on the passage time of food through the whole stomach. The latter hypothesis was indirectly tested by assuming, based on previous evidence from other vertebrates, that the greater the size of the stomach and/or the energy content of prey, the greater the delay of food passage. We found no compelling evidence that chamber colonization was sequential, or related to chamber size in any species. However, the distribution of P. gastrophilus was significantly more anteriad when the host species had larger stomachs and, particularly, when hosts fed on prey with higher caloric content. Accordingly, the stomach distribution of P. gastrophilus at this scale seems to be passively driven by features of the diet and digestive physiology of each host species. This study provides a general framework to formulate null hypotheses in future studies on microhabitat choice by parasites. PMID- 16719958 TI - A tryptophan amphiphilic tetramerization domain-containing acetylcholinesterase from the bovine lungworm, Dictyocaulus viviparus. AB - Acetylcholine (ACh) is one of an array of neurotransmitters used by invertebrates and, analogous to vertebrate nervous systems, acetylcholinesterase (AChE) regulates synaptic levels of this transmitter. Similar to other invertebrates, nematodes possess several AChE genes. This is in contrast to vertebrates, which have a single AChE gene, transcripts of which are alternatively spliced to produce different types of the enzyme which vary at their C-termini. Parasitic nematodes have a repertoire of AChE genes which include those encoding neuromuscular AChEs and those genes which code for secreted AChEs. The latter proteins exist as soluble monomers released by the parasite during infection and these AChE are distinct from those enzymes which the nematodes use for synaptic transmission in their neuromuscular system. Thus far, Dictyocaulus viviparus is the only animal-parasitic nematode for which distinct genes that encode both neuromuscular and secreted AChEs have been defined. Here, we describe the isolation and characterization of a cDNA encoding a putative neuromuscular AChE from D. viviparus which contains a tryptophan amphiphilic tetramerization (WAT) domain at its C-terminus analogous to the common 'tailed' AChE form found in the neuromuscular systems of vertebrates and in the ACE-1 AChE from Caenorhabditis elegans. This enzyme differs from the previously isolated, D. viviparus neuromuscular AChE (Dv-ACE-2), which is a glycosylphosphatidylinositol-anchored variant analogous to vertebrate 'hydrophobic' AChE. PMID- 16719959 TI - Geographical variations in infectivity and susceptibility in the host-parasite system Schistosoma mansoni/Biomphalaria glabrata: no evidence for local adaptation. AB - We investigated local adaptation in the spatially structured natural Biomphalaria glabrata/Schistosoma mansoni host-parasite system in the marshy forest focus of Guadeloupe using cross-transplantation experiments. We demonstrated strong and highly significant variations in susceptibility/infectivity of host and parasite populations, respectively, but found no evidence of local adaptation neither for S. mansoni nor for B. glabrata. Environmental as well as genetic factors are discussed to explain susceptibility/infectivity variations between both host and parasite populations. The absence of local adaptation is discussed in relation to the metapopulation dynamics of both host and parasite, in particular their relative rates of dispersal at the scale under scrutiny. Our study constitutes the first cross-transplantation experiment concerning this host-parasite system of which both hosts and parasites came directly from the wild, excluding laboratory generations and experimental host passages. PMID- 16719960 TI - Comparison of the transmissibility of Trypanosoma congolense strains, isolated in a trypanosomiasis endemic area of eastern Zambia, by Glossina morsitans morsitans. AB - Transmission experiments were conducted to compare the transmissibility of genetically different Trypanosoma congolense (Savannah subgroup) strains isolated from cattle in a trypanosomiasis endemic area of eastern Zambia. A total of 17 strains were compared. Three strains were extremely virulent with a short pre patent period, high parasitaemia and a short median survival time (between 5 and 9 days) in mice. The remainder of the strains belonged to the moderate (6 strains) or low (8 strains) virulence categories with median survival times between 10 and 30 days and >30 days, respectively. Batches of 40 teneral Glossina morsitans morsitans (Diptera: Glossinidae) were offered a single bloodmeal on mice infected with one of those strains. Flies were dissected to determine their infection status 21 days later. The proportion of flies with procyclic and metacyclic infections differed significantly between trypanosome strains and were significantly higher in flies infected with extremely virulent strains (P=0.033 and P=0.016 for the differences in the procyclic infection rate of strains with moderate and low virulence, respectively and P=0.005 and P=0.019 for the differences in the metacyclic infection rate of strains with moderate and low virulence, respectively). On the other hand, moderately virulent strains had, in general, higher procyclic and metacyclic infection rates compared to low virulent strains. But the differences were not significant (P>0.05). The outcome of those experiments shows clear differences in transmissibility of trypanosome strains associated with their virulence. This observation confirms the theory for the evolution and maintenance of virulence in a parasite population and may explain the persistence of virulent trypanosome strains in a susceptible host population. PMID- 16719961 TI - The functional and developmental organization of cognitive developmental sequences. AB - This study examines the organization and development of 5 domains of reasoning (categorical, quantitative, spatial, causal, and propositional) and the construct validity of a test designed to measure development from early adolescence to early adulthood. The theory underlying the test is first summarized and the conceptual design of the test is then illustrated. Each domain was addressed by tasks tapping abilities known to be acquired in this age period. The test was administered to 629 adolescents ranging in age from 12 to 18 years. Confirmatory factor analysis validated the 5 domains of reasoning and revealed a common factor underlying all domains. The Rasch model was used to scale the items and specify the reliability of the test across the whole sample and within different groups of participants (female, male, students of gymnasium, and students of lyceum). This model showed that the test is highly reliable and invariant across groups. Cluster analysis and the saltus model were applied to uncover successive developmental stage-like levels of difficulty and showed the presence of five such levels. The procedural and representational characteristics of these levels were also specified and their implications for developmental and cognitive theory were discussed. PMID- 16719962 TI - Foreign-grammar acquisition while watching subtitled television programmes. AB - BACKGROUND: Past research has shown that watching a subtitled foreign movie (i.e. foreign language in the soundtrack and native language in the subtitles) leads to considerable foreign-language vocabulary acquisition; however, acquisition of the grammatical rules has failed to emerge. AIMS: The aim of this study was to obtain evidence for the acquisition of grammatical rules in watching subtitled foreign movies. Given an informal context, younger children were predicted to outperform older children in acquiring a foreign language; however, older children will take more advantage of explicit instruction compared with younger children. SAMPLE: In Experiment 1, 62 sixth-graders from a primary school and 47 sixth-graders from a secondary school volunteered to participate. The participants in Experiment 2 were 94 sixth-graders from primary schools and 84 sixth-graders from secondary schools. METHOD: The two experiments manipulated the instructions (incidental- vs. intentional-language learning). Moreover, before the experiments began, some participants explicitly received some of the foreign grammatical rules (presented rules), while the movie contained cases of presented rules as well as cases of rules which had to be inferred (not-presented rules). RESULTS: Rule acquisition through the movie only was not obtained; there was a strong effect of advance rule presentation but only on the items of presented rules, particularly among the older participants. CONCLUSIONS: Contrary to vocabulary, grammar may be too complicated to acquire from a rather short movie presentation. PMID- 16719963 TI - The effects of different forms of feedback on fuzzy and verbatim memory of science principles. AB - BACKGROUND: Previous models of the effects of feedback account for lower-order learning outcomes but do not adequately describe experimental findings for higher order learning. AIMS: Based on a connectionist model of feedback effects, this investigation aims to show that feedback that allows only one learner response facilitates proposition-specific verbatim encoding, while feedback that requires the learner to try again on error facilitates relational fuzzy encoding. Sample and methods. Volunteer high school students were randomly assigned to one of 5 print-based lesson treatments that consisted of four science expository texts with adjunct inference-level questions covering science principles. The five treatments included delayed feedback, single-try immediate feedback, multiple-try immediate feedback, and two control treatments, questions without feedback, and text only. A post-test given 5 days after instruction was designed to measure both verbatim and fuzzy outcomes. RESULTS: Multiple-try immediate feedback was best for paraphrased post-test questions (fuzzy) and worse for verbatim post-test questions. CONCLUSIONS: Fuzzy trace theory complements a connectionist model of feedback, and may provide a fruitful approach for describing the effects of feedback on different learning outcomes. PMID- 16719964 TI - Teachers' and students' verbal behaviours during cooperative and small-group learning. AB - BACKGROUND: Teachers play a critical role in promoting interactions between students and engaging them in the learning process. This study builds on a study by Hertz-Lazarowitz and Shachar (1990) who found that during cooperative learning teachers' verbal behaviours were more helpful to and encouraging of their students' efforts while during whole-class instruction, their verbal behaviours tended to be more authoritarian, rigid, and impersonal. AIM: This study seeks to determine if teachers who implement cooperative learning engage in more facilitative learning interactions with their students than teachers who implement group work only. The study also seeks to determine if students in the cooperative groups model their teachers' behaviours and engage in more positive helping interactions with each other than their peers in the group work groups. SAMPLES: The study involved 26 teachers and 303 students in Grades 8 to 10 from 4 large high schools in Brisbane, Australia. METHODS: All teachers agreed to establish cooperative, small-group activities in their classrooms for a unit of work (4 to 6 weeks) once a term for 3 school terms. The teachers were audiotaped twice during these lessons and samples of the students' language, as they worked in their groups, were also collected at the same time. RESULTS: The results show that teachers who implement cooperative learning in their classrooms engage in more mediated-learning interactions and make fewer disciplinary comments than teachers who implement group work only. Furthermore, the students model many of these interactions in their groups. CONCLUSIONS: The study shows that when teachers implement cooperative learning, their verbal behaviour is affected by the organizational structure of the classroom. PMID- 16719965 TI - Academic emotions from a social-cognitive perspective: antecedents and domain specificity of students' affect in the context of Latin instruction. AB - This study concentrates on two assumptions of a social-cognitive model outlining the development of academic emotions (emotions directly linked to learning, classroom instruction, and achievement), namely on their antecedents and domain specific organization. Our sample consisted of 200 students from Grades 7 to 10. Proposed relationships concerning the antecedents of academic emotions were tested in the context of Latin language instruction. Correlational analyses substantiated our assumptions concerning the relationships between academic emotions, students' cognitions, and aspects of the social environment. The mediating mechanisms proposed in the model were also confirmed using linear structural equation modelling. Subjective control- and value-related cognitions were found to mediate the relationship between aspects of the social environment and students' emotional experience. Our results further suggest that academic emotions are largely organized along domain-specific lines, with the degree of domain specificity varying according to the emotion in question. Implications for research and practice are discussed. PMID- 16719966 TI - Individual differences in children's understanding of inversion and arithmetical skill. AB - Background and aims. In order to develop arithmetic expertise, children must understand arithmetic principles, such as the inverse relationship between addition and subtraction, in addition to learning calculation skills. We report two experiments that investigate children's understanding of the principle of inversion and the relationship between their conceptual understanding and arithmetical skills. SAMPLE: A group of 127 children from primary schools took part in the study. The children were from 2 age groups (6-7 and 8-9 years). METHODS: Children's accuracy on inverse and control problems in a variety of presentation formats and in canonical and non-canonical forms was measured. Tests of general arithmetic ability were also administered. RESULTS: Children consistently performed better on inverse than control problems, which indicates that they could make use of the inverse principle. Presentation format affected performance: picture presentation allowed children to apply their conceptual understanding flexibly regardless of the problem type, while word problems restricted their ability to use their conceptual knowledge. Cluster analyses revealed three subgroups with different profiles of conceptual understanding and arithmetical skill. Children in the 'high ability' and 'low ability' groups showed conceptual understanding that was in-line with their arithmetical skill, whilst a 3rd group of children had more advanced conceptual understanding than arithmetical skill. CONCLUSIONS: The three subgroups may represent different points along a single developmental path or distinct developmental paths. The discovery of the existence of the three groups has important consequences for education. It demonstrates the importance of considering the pattern of individual children's conceptual understanding and problem-solving skills. PMID- 16719967 TI - Reading errors in first- and second-grade readers of a shallow orthography: evidence from Spanish. AB - BACKGROUND: Studies focusing on reading errors can help to understand how children learn to read and to structure key components of reading instruction. However, no prior studies have examined which letters might be a greater source of difficulty for beginning readers in Spanish. AIMS: First, to examine the pattern of reading errors in beginning readers in a shallow orthography like Spanish with a particular focus on both the context-dependant (c, g, r) and the visual similar (b, d, g, p, q) consonants. (The dual-route cascade model is used as a theoretical framework for predictions.) Second, to examine the effect of visual similarity on orientation reversals, using upper and lower case letters. SAMPLE: Ninety Spanish-speaking children (45 in Grade 1 and 45 in Grade 2) participated in the study. METHOD: Children were required to read aloud both words and non-words. Seven categories of errors were examined. To clarify the influence of visual similarity of easily confused consonants (b, d), upper and lower case letters were used. RESULTS: We found that most reading errors (as a function of opportunity) were due to context-dependant consonants (c, g, r), and a small proportion were due to the remaining consonants, or even the visually similar consonants. Further, visual similarity of reversible letters had a significant effect on orientation reversals. Finally, there was a significant influence of factors such as syllable frequency on the number of word errors. CONCLUSIONS: The implications of these findings for early reading instruction and future research are discussed. PMID- 16719968 TI - Evidence for the effectiveness of the Early Literacy Support programme. AB - BACKGROUND: It is widely recognized that effective interventions for poor reading involve training in phoneme awareness and letter-sound knowledge, linked in the context of reading books. From the applied perspective, it is important to gather data on the effectiveness of different forms of implementation of literacy support within this framework. AIM: We evaluate the effectiveness of the UK Early Literacy Support (ELS) programme (Department for Education and Skills [DfES], 2001) relative to a programme of Reading Intervention based on 'sound linkage' (Hatcher, Hulme, & Ellis, 1994). SAMPLE: The sample comprised 128 6-year-old children, from 16 primary schools in a Local Education Authority (LEA) in the north of England. METHOD: The children were nominated as in need of special help by their class teachers and allocated to one of the two programmes. RESULTS: After controlling for a difference in spelling ability at the start of the intervention, it was found that both groups of children made equivalent and significant gains in reading and spelling that were maintained at follow-up. The standardized scores were in the average range. Regression analyses confirmed the importance of initial reading ability as well as letter identification, phoneme manipulation, and sound linkage in predicting progress in learning to read and to spell. CONCLUSION: The ELS programme provides a cost effective method of boosting 6-year-old children's reading to an average level. PMID- 16719969 TI - Situational state balances and participation motivation in youth sport: a reversal theory perspective. AB - BACKGROUND: Reversal theory (Apter, 1982, 1989, ) is one of the motivational frameworks which attempts to examine human subjective experiences and behaviours. There are four dyads of metamotivational states (telic-paratelic, conformist negativistic, autic-alloic, and mastery-sympathy) and individuals may prefer to be in one rather than the other of a dyad of states in a specific context such as sport participation (i.e. situational state balances). AIMS: The purpose of this study was to examine the relationship between situational state balances and motives for sport and physical activity participation in adolescents using the theoretical framework of reversal theory. Their participation motives and the interacting factors of their situational state balances, gender, and level of participation were examined. SAMPLE: Secondary school students (N=1,235) aged about 14 to 20 years who participated in competitive or recreational sport completed the Participation Motivation Inventory (Gill, Gross, & Huddleston, 1983) to assess their motives for sport and physical activity participation, and the Apter Motivational Style Profile (Apter International, 1999) to assess their situational state balances. METHODS: Factor analysis of the participation motives yielded factors to which MANOVAs and ANOVAs were applied with situational state balance, gender, and participation level as independent variables. RESULTS: Factor analysis resulted in seven motive factors: status, team/friend, excitement/challenge, skill, energy release, fitness, and situational factors. MANOVAs and ANOVAs indicated significant differences in the sport motives between the situational state balances, genders, and levels of participation, and between pairs of situational state balance groups in males and females of competitive and recreational level. The autic-alloic dyad produced the strongest motive strength differences of the four pairs. CONCLUSIONS: The variable of situational state balances is significantly linked with participation motives in sport. PMID- 16719970 TI - Observation of children with attention-deficit hyperactivity (ADHD) problems in three natural classroom contexts. AB - BACKGROUND: Observation studies of students with attention-deficit hyperactivity disorder (ADHD) problems in natural classroom situations are costly and relatively rare. AIMS: The study enquired how teacher ratings are anchored in actual student classroom behaviours, and how the behaviour of children with ADHD problems differs from their classmates. The authors attempted to broaden the usual focus on disruptive and inattentive behaviours to elucidate the role of various on-task behaviours, as well as considering differences between classroom contexts. SAMPLE: DSM-III-R criteria were used in conjunction with a teacher rating scale to select a sample of 55 students with ADHD problems, and 55 matched controls from a population of 569 primary school students. METHOD: Students were observed in their natural classrooms using the Munich Observation of Attention Inventory (MAI; Helmke, 1988). Correlations between teacher reports and observation codes were computed, and systematic differences between students with ADHD problems and controls in different classroom contexts were examined using a generalized linear mixed model (GLMM). RESULTS: Global teacher reports showed moderate to strong correlations with observed student behaviours. Expected on task behaviour demonstrated the strongest relationship (r>-.70) with teacher reports. As hypothesized, the children with ADHD were more disruptive and inattentive than their matched peers. They were also less often inconspicuous on task as expected by their teachers. However, their behaviour was assigned to two other on-task categories more often than their peers, and this raised their total on-task behaviour to over 66%. Situational differences were found for all codes as well, which mostly affected all students in a similar way, not just students with ADHD. CONCLUSIONS: ADHD related behaviours are pervasive across the classroom situations coded. Teachers appear to distinguish between desirable and undesirable on-task behaviours. Nevertheless, assisting students with ADHD problems requires shaping both. Future studies need to include more differentiated codes for various types of on-task behaviours and also need to code the lesson context concurrently. PMID- 16719971 TI - Confirmatory factor analysis of the Approaches to Teaching Inventory. AB - BACKGROUND: The Approaches to Teaching Inventory (ATI) was developed to explore the relationship between students' approaches to learning, and teachers' approaches to teaching, in the physical sciences in higher education. It is increasingly being used in other contexts. AIM: To analyse the factor structure of the ATI. METHOD: Confirmatory factor analysis. SAMPLE: Inventory responses from over 1,000 university teachers, from 4 different countries and a wide range of disciplines, experience, and teaching level (year of students' study) were used as the sample. RESULTS: Cronbach's alpha values of the 2-scale inventory were acceptable for scale reliability, but not for the 4-scale structure. A good fit to the intended 2-factor structure was obtained by confirmatory factor analysis. Both Information transmission/teacher-focused and conceptual change/student-focused scales had well identified intention and strategy subscales. CONCLUSION: The ATI has been developed as a relational indicator of 2 dimensions of approaches to teaching. It is not intended for use in gathering a full, rich self-report of teaching, or in non-relational contexts. However, it has now been used in a number of different contexts, including different subject areas and cultures. There is evidence in this paper that in those contexts its factor structure is, in the main, consistent with the original design. This analysis also suggests ways that the ATI could be improved. PMID- 16719976 TI - A factor analysis of the Wechsler Adult Intelligence Scale 3rd Edition (WAIS-III) in a low IQ sample. AB - OBJECTIVES: Previous factor analytic studies of the WAIS, WAIS-R and WAIS-III used standardization samples, (representing the general population), clinical populations (e.g. psychiatric) and 'non-clinical' groups (e.g. older adults). These studies endorsed the reliability of the scales in such samples and supported theoretical models of intelligence. The WAIS-III (1997) includes four Indexes based on factor analysis, which provide clinically useful information to practitioners, but have not been validated in a low IQ population. However, the WAIS-III is often administered to individuals with suspected or proven low IQ, as it offers service providers, legislators and the Department of Health with a reliable measure of IQ. The aim of this study was to investigate the factor structure of the WAIS-III in a low IQ sample. METHODS: WAIS-III data was collected from assessments carried out in routine clinical practice from individuals with a full scale IQ of 74 or below (n=105). The data were subjected to factor analysis, using two types of factor analysis: principal axis factoring and principal components analysis. Orthogonal and oblique rotations were applied to the analyses. RESULTS: Only one robust solution could be extracted. This contained two factors, analogous to traditional verbal and performance sub scales. CONCLUSIONS: This study does not support the four-factor solution which underlies WAIS-III index scores in a low IQ population. PMID- 16719977 TI - Transforming between Beck Depression Inventory and CORE-OM scores in routine clinical practice. AB - OBJECTIVES: The Clinical Outcomes in Routine Evaluation - Outcome Measure (CORE OM) and the Beck Depression Inventory (BDI) are routinely used to assess emotional problems. It would be helpful to be able to compare scores when only one of the measures is available. We investigated the relationship between the measures and produced translation tables. METHODS: Level of agreement between CORE-OM and BDI-I was assessed for 2,234 clients who had completed both measures at referral for routine secondary care. Tables for predicting between the measures were constructed using several methods, including non-linear regression and non-parametric smoothing. Results were cross-validated on a separate sample of 326 clients. RESULTS: High correlations between the measures were obtained (r=.862 for female clients; r=.855 for male clients). Accuracy in predicting caseness is higher than predicting depression levels. CONCLUSIONS: CORE-OM and BDI can be compared in routine clinical settings with acceptable accuracy. PMID- 16719978 TI - The effect of a negative mood priming challenge on dysfunctional attitudes, explanatory style, and explanatory flexibility. AB - Ninety-seven undergraduates, 48 of whom had a history of self-reported major depression, completed measures of mood and cognitive style (e.g. explanatory style, explanatory flexibility, dysfunctional attitudes) prior to and directly after a negative mood priming challenge that consisted of listening to sad music and thinking about an upsetting past event. Eighteen of the previously depressed participants endorsed baseline levels of depression, explanatory style for negative events, and dysfunctional attitudes higher than levels reported by never depressed participants or euthymic participants with a history of depression. All three groups (never depressed participants, dysphoric participants with a history of depression, euthymic participants with a history of depression) demonstrated increases in dysphoria and dysfunctional attitudes in response to the negative mood priming challenge. Dysphoric participants with a history of depression, but not the other two groups, evidenced modest increases in explanatory style following the negative mood priming challenge. Finally, euthymic participants with a history of depression, but not the other two groups, evidenced drops in explanatory flexibility. Findings from the present study suggest that the cognitive theories of depression may benefit from examining both cognitive content and cognitive flexibility when assessing risk for depression. PMID- 16719980 TI - Risk factors for psychological distress following injury. AB - OBJECTIVES: To identify predictors of psychological morbidity among injured patients admitted to an Emergency Department (ED). DESIGN: A prospective cohort study. PARTICIPANTS: Participants were consecutive male ED attenders. 210 (97.7%) patients consented to participate. At one month, 128 (61.0%) responded, at six months, 114 (54.3%), at eighteen months 96 (45.7%). MAIN OUTCOME MEASURES: Measures immediately following injury were the Hospital Anxiety and Depression Scale, the Eysenck Personality Questionnaire and the McGill pain questionnaire. Recovery at one month was recorded using the SF-36 Health Survey, COPE scale, Perceived Stress Scale and Revised Impact of Events Scale. At six and eighteen months outcome was measured using the General Health Questionnaire (28 items) and Revised Impact of Events Scale. Multivariate analysis identified pre-morbid, accident-related and recovery factors influencing outcome at six and eighteen months. RESULTS: The strongest predictors of outcome were initial levels of anxiety and depression, prior history of mental health problems, early PTSD symptoms and involvement in litigation. These factors predicted between 40-60% of the variance at six months (p<0.001), and 50-60% of the variance in psychological distress at eighteen months (p<0.001). CONCLUSION: Factors identifying individuals at-risk from psychological distress following injury include those related to the immediate response and the recovery phases of injury. Further development is needed to convert identified predictors into a comprehensive screening tool for clinical use. PMID- 16719979 TI - Challenging behaviour and learning disabilities: the relationship between expressed emotion and staff attributions. AB - OBJECTIVES: Expressed emotion (EE) and attributions towards challenging behaviour (CB) were explored amongst a group of staff working within a residential and day service placement for people with learning disabilities. DESIGN: Using a cross sectional related-samples design, EE and attributions were measured amongst all staff working with one client with CB, and one client without CB. METHODS: Fifteen staff members completed the attributional questionnaire and the 5-minute speech sample (FMSS) to allow for EE ratings concerning staff relationships with two clients. One client exhibited CB, while the other did not, giving 2 samples. Attributional and EE ratings for each group were compared. This study did not employ vignette methodology. RESULTS: Staff working with a client with learning disabilities and CB attributed the CB as internal to the client and controllable by the client. Staff reported high levels of EE and made more critical comments towards the client with CB as compared with the client without CB. Furthermore, staff who reported high EE attributed CB as internal to the client and controllable by the client. CONCLUSIONS: Staff working with a client with challenging behaviour appeared to be making the 'fundamental attribution error'. The relationship between expressed emotion and attribution theory is discussed along with the methodological benefits of not relying on vignette methodology in research that examines challenging behaviour. PMID- 16719981 TI - Attributional style, defensive functioning and persecutory delusions: symptom specific or general coping strategy? AB - OBJECTIVE: Previous research has suggested that individuals with persecutory delusions use an exaggerated self-serving bias to protect themselves from real or delusional threats to their underlying self-concept. However, the research to date has been inconsistent thereby limiting the conclusion that can be inferred. A possible explanation for these discrepant findings may be due to the use of measures with questionable validity and reliability such as the Attributional Style Questionnaire and the Rosenberg Self-Esteem Inventory. The present study aims to re-examine this theory using an improved methodology to determine whether defensive functioning is a defining feature of persecutory delusions. DESIGN AND METHODS: Thirty-five participants suffering from recent onset psychosis were assessed in a cross-sectional design. Three tests of attributional style were used, two overt measures (Attributional Style Questionnaire, ASQ; and the Internal Personal and Situational Attributions Questionnaire, IPSAQ) and one covert measure (Pragmatic Inference Test). Self-esteem was measured using the Rosenberg Self-Esteem Inventory (SEI) and a semi-structured interview (Self Evaluation and Social Support interview - schizophrenia version). RESULTS: A self serving bias (SSB) was found in the total sample using the ASQ but not specifically in participants with persecutory delusions. In addition, the SSB was unrelated to levels of paranoia on any measure of attributional style. An underlying depressive attributional style was found in both subjects with and without persecutory delusions, but was not associated with levels of paranoia. Lower positive and higher negative self-esteem was associated with increased paranoia and was associated with a SSB. CONCLUSION: The results indicate that the attributional biases observed in individuals with persecutory delusions are not symptom-specific as previously suggested. The presence of an attributional, SSB was associated with lower psychotic experiences. The theoretical implications of the results will be discussed. PMID- 16719982 TI - Early intervention for relapse in schizophrenia: impact of cognitive behavioural therapy on negative beliefs about psychosis and self-esteem. AB - OBJECTIVES: The study aimed to test two hypotheses. Firstly, that participants who relapsed during the 12-month follow-up period of our randomized controlled trial, would show increased negative beliefs about their illness and reduced self esteem, in comparison to the non-relapsed participants. Secondly, that cognitive behavioural therapy (CBT) for early signs of relapse would result in a reduction in negative beliefs about psychosis and an improvement in self-esteem at 12 months. DESIGN AND METHODS: A total of 144 participants with schizophrenia or a related disorder were randomized to receive either treatment as usual (TAU; N=72) or CBT (N=72). Participants completed the Personal Beliefs about Illness Questionnaire (PBIQ; Birchwood, Mason, MacMillan, & Healy, 1993) and the Rosenberg Self-Esteem Scale (RSES; Rosenberg, 1965) at entry, 3 months, 6 months, and 12 months. RESULTS: At 12 months, relapsers showed greater increase in scores for PBIQ entrapment compared with non-relapsers. In addition, after controlling for baseline covariates (treatment group and PBIQ self versus illness), relapsers also showed greater increase in scores for PBIQ self versus illness at 12 months. Furthermore, in comparison to treatment as usual, participants who received CBT showed greater improvement in PBIQ loss and in Rosenberg self-esteem. CONCLUSIONS: The study provides evidence that relapse is associated with the development of negative appraisals of entrapment and self-blame (self vs. illness). In addition, this is the first study to show that CBT reduces negative appraisals of loss arising from psychosis and improvements in self-esteem. Implications for future research and treatment are discussed. PMID- 16719983 TI - Understanding of basic financial concepts among adults with mild learning disabilities. AB - OBJECTIVE: In the context of recent legal and social policy developments, which encourage clinicians to support vulnerable adults in making their own financial decisions, we explored understanding of basic financial concepts among adults with mild learning disabilities. METHOD: Using the literature about typically developing children, five measures entailing identification and ordering tasks were devised and were used to examine understanding of quantity, numbers and money among 30 men and women using specialist clinical services (mean FSIQ = 61.80; SD=10.59). RESULTS: The concept of quantity was easier to understand than those of numbers and money, and task difficulties increased with the magnitudes involved. The different types of conceptual understanding were closely related. CONCLUSIONS: These measures may be useful for analysing difficulties in, and informing interventions to maximize, independent financial decision making. PMID- 16719984 TI - The effects of rumination and distraction on over-general autobiographical memory retrieval during social problem solving. AB - OBJECTIVES: We examined the effects of rumination and distraction on over-general autobiographical memory (AM) retrieval during social problem solving (SPS), as well as SPS performance in terms of means and effectiveness. DESIGN AND METHODS: After undergoing a rumination or distraction manipulation, dysphoric and non dysphoric participants performed a SPS task and reported the memories retrieved during SPS. RESULTS: The dysphoric ruminators performed significantly less effectively on the SPS task than the other groups. However, the dysphoric ruminators did not differ from the dysphoric distracters on the numbers of categoric memories retrieved during SPS although they retrieved significantly more categoric memories than the non-dysphoric distracters and non-dysphoric ruminators. For all groups, the effectiveness score was significantly negatively correlated with categoric memories retrieved during SPS. CONCLUSION: The results demonstrated that in a dysphoric sample, rumination impaired SPS performance but did not influence categoric AM retrieval during SPS. The results suggested that in a dysphoric sample, rumination might affect SPS performance by affecting processes like initiation and motivation rather than AM retrieval during SPS. PMID- 16719985 TI - Dependent personality traits and information processing: assessing the interpretation of ambiguous information using the Thematic Apperception Test. AB - OBJECTIVES: This study was designed to investigate interpretation bias in people with dependent personality traits. METHOD: Eight Thematic Apperception Test (TAT) cards were administered to participants (N=56) who scored high or low on DSM-III R dependent personality pathology. Two independent judges rated the TAT stories using a rating list based on the cognitive model of the dependent and paranoid personality disorder. RESULTS: Controlling for self-esteem, the dependent interpretation bias appeared to be specific for dependent personality pathology. SEM analysis supported a mediation model in which beliefs mediate the relationship between DSM-III-R traits and interpretation bias. CONCLUSIONS: The findings in this study support the hypothesis that people with dependent traits are characterized by a schema-related interpretation bias and that this bias is mediated by dependent beliefs. PMID- 16719986 TI - Tolerance to microbial TLR ligands: molecular mechanisms and relevance to disease. AB - Many host cell types, including endothelial and epithelial cells, neutrophils, monocytes, natural killer cells, dendritic cells and macrophages, initiate the first line of defense against infection by sensing conserved microbial structures through Toll-like receptors (TLRs). Recognition of microbial ligands by TLRs induces their oligomerization and triggers intracellular signaling pathways, leading to production of pro- and anti-inflammatory cytokines. Dysregulation of the fine molecular mechanisms that tightly control TLR signaling may lead to hyperactivation of host cells by microbial products and septic shock. A prior exposure to bacterial products such as lipopolysaccharide (LPS) may result in a transient state of refractoriness to subsequent challenge that has been referred to as 'tolerance'. Tolerance has been postulated as a protective mechanism limiting excessive inflammation and preventing septic shock. However, tolerance may compromise the host's ability to counteract subsequent bacterial challenge since many septic patients exhibit an increased incidence of recurrent bacterial infection and suppressed monocyte responsiveness to LPS, closely resembling the tolerant phenotype. Thus, by studying mechanisms of microbial tolerance, we may gain insights into how normal regulatory mechanisms are dysregulated, leading ultimately to microbial hypo-responsiveness and life-threatening disease. In this review, we present current theories of the molecular mechanisms that underlie induction and maintenance of 'microbial tolerance', and discuss the possible relevance of tolerance to several infectious and non-infectious diseases. PMID- 16719987 TI - Compartmentalization of the inflammatory response in sepsis and SIRS. AB - Sepsis and systemic inflammatory response syndrome (SIRS) are associated with an exacerbated production of both pro- and anti-inflammatory mediators that are mainly produced within tissues. Although a systemic process, the pathophysiological events differ from organ to organ, and from organ to peripheral blood, leading to the concept of compartmentalization. The nature of the insult (e.g. burn, hemorrhage, trauma, peritonitis), the cellular composition of each compartment (e.g. nature of phagocytes, nature of endothelial cells), and its micro-environment (e.g. local presence of granulocyte-macrophage colony stimulating factor [GM-CSF] in the lungs, low levels of arginine in the liver, release of endotoxin from the gut), and leukocyte recruitment, have a great influence on local inflammation and on tissue injury. High levels of pro inflammatory mediators (e.g. interleukin-1 [IL-1], tumor necrosis factor [TNF], gamma interferon [IFN-gamma], high mobility group protein-1 [HMGB1], macrophage migration inhibitory factor [MIF]) produced locally and released into the blood stream initiate remote organ injury as a consequence of an organ cross-talk. The inflammatory response within the tissues is greatly influenced by the local delivery of neuromediators by the cholinergic and sympathetic neurons. Acetylcholine and epinephrine contribute with IL-10 and other mediators to the anti-inflammatory compensatory response initiated to dampen the inflammatory process. Unfortunately, this regulatory response leads to an altered immune status of leukocytes that can increase the susceptibility to further infection. Again, the nature of the insult, the nature of the leukocytes, the presence of circulating microbial components, and the nature of the triggering agent employed to trigger cells, greatly influence the immune status of the leukocytes that may differ from one compartment to another. While anti-inflammatory mediators predominate within the blood stream to avoid igniting new inflammatory foci, their presence within tissues may not always be sufficient to prevent the initiation of a deleterious inflammatory response in the different compartments. PMID- 16719988 TI - Endotoxin evaluation of eleven lipopolysaccharides by whole blood assay does not always correlate with Limulus amebocyte lysate assay. AB - More than 90% of all publications on endotoxin were carried out with endotoxins (lipopolysaccharide, LPS) from enterobacteriaceae. We compared the immune stimulatory potency of 11 different LPSs using human whole blood incubations. While the majority of LPSs induced cytokine release equipotently, a 1,000-fold more LPS from Pseudomonas aeruginosa or Vibrio cholerae was still less potent in inducing TNF, IL-1 beta, IL-10 and IFN-gamma though it potently induced nanogram quantities IL-8. All LPSs tested, regardless of the micro-organism, showed Toll like receptor (TLR)4-dependence, except for the LPSs from P. aeruginosa and V. cholerae, which were both TLR4- and TLR2-dependent. Interestingly, UV-inactivated P. aeruginosa bacteria, although Gram-negative, also showed TLR2- and TLR4 dependence. Re-purification of commercial LPS preparations by phenol re extraction led to a complete loss of the TLR2 dependency, indicating contamination with lipoproteins. In the Limulus amebocyte lysate assay, often performed to exclude contamination in purified water likely to originate from P. aeruginosa, P. aeruginosa LPS was only 2-fold less potent than LPS from S. abortus equi or the assay standard LPS from E. coli. This results in an overestimation of pyrogenic burden by a factor of 500 in the sample when compared with the biological activity of highly purified P. aeruginosa LPS in human whole blood. PMID- 16719989 TI - Impaired response of VLDL lipid and apoB secretion to endotoxin in the fasted rat liver. AB - Bacterial infection elicits hypertriglyceridemia attributed to increased hepatic production of very low-density lipoprotein (VLDL) particles and decreased peripheral metabolism. The mechanisms underlying VLDL overproduction in sepsis are as yet unclear, but seem to be fed/fasted state-dependent. To learn more about this, we investigated hepatocytes isolated from fasted rats, made endotoxic by 1 mg/kg lipopolysaccharide (LPS) injection, for their ability to secrete the VLDL protein and lipid components. The results were then related to lipogenesis markers and expression of genes critical to VLDL biogenesis. Endotoxic rats showed increased levels of serum VLDL-apoB (10-fold), -triglyceride (2-fold), and -cholesterol (2-fold), whereby circulating VLDL were lipid-poor particles. Similarly, VLDL-apoB secretion by isolated endotoxic hepatocytes was approximately 85% above control, whereas marginal changes in the output of VLDL lipid classes occurred. This was accompanied by a substantial rise in apoB and a moderate rise in MTP mRNA levels, but with basal de novo formation and efficiency of secretion of triglycerides, cholesterol and cholesteryl esters. These results indicate that during periods of food restriction, endotoxin does not enhance lipid provision to accomplish normal lipidation of overproduced apoB molecules, though this does occur to a sufficient extent to pass the proteasome checkpoint and secretion of lipid-poor, type 2 VLDL takes place. PMID- 16719990 TI - CORESS - a confidential reporting system for surgery. PMID- 16719993 TI - Consent in surgery. AB - A review of consent for surgery is timely. As the length of surgeons' training diminishes, despite the increasing interest in the content of the surgical curriculum, the law governing the process of gaining consent has been given scant attention. The advent of non-medically qualified surgical practitioners raises questions about the breadth of knowledge that is required to ensure that valid consent is obtained. Consent is as fundamental as any other basic principle on which surgical practice relies, and its use in patient care is a clinical skill. The 'traditional' approach to consent contained some negative elements. A doctor who was incapable of performing the proposed operation often obtained consent. In a genuine attempt to protect patients from anxiety, the rare-but-grave potential complications were sometimes not discussed. There was uncertainty about what should properly be disclosed, compounded by conflicting messages from the courts. The consent was sometimes taken from people who were ineligible to provide it. These could be viewed as aberrations, and some persist. Having clarified the necessity for consent, this review concludes that it should be obtained by the operating surgeon. The threshold for interventions that need formal consent is discussed, together with the legal tests for capacity. In considering the recent law, it becomes clear that any potential complication that the reasonable patient would need to take into consideration before deciding to give their consent is one that should be disclosed. PMID- 16719992 TI - The management of incisional hernia. AB - Many thousand laparotomy incisions are created each year and the failure rate for closure of these abdominal wounds is between 10-15%, creating a large problem of incisional hernia. In the past many of these hernias have been neglected and treated with abdominal trusses or inadequately managed with high failure rates. The introduction of mesh has not had a significant impact because surgeons are not aware of modern effective techniques which may be used to reconstruct defects of the abdominal wall. This review will cover recent advances in incisional hernia surgery which affect the general surgeon, and also briefly review advanced techniques employed by specialist surgeons in anterior abdominal wall surgery. PMID- 16719994 TI - Coronary artery stenosis. AB - The near exponential rise in percutaneous coronary intervention(PCI) in the treatment of patients with coronary artery disease and the consequent decline in referral of patients for coronary artery bypass grafting (CABG) has lead to a crisis in cardiac surgery. Is CABG, one of the most successful and widely applied surgical procedures, about to follow surgery for peptic ulcer disease into obsolescence? The question has serious implications for service provision and training as well as for informed patient consent. Keith Dawkins puts the case for PCI and gives a very clear and concise account of its inexorable rise to pre eminence. David Taggart has taken on the Goliath of interventional cardiology and its associated industry by persuasively marshalling the data from the evidence base which strongly favours surgery in triple vessel and left main coronary disease. He points to the lack of long-term results and also to the weaknesses of many of the comparative studies so far published. His arguments have been positively received on both sides of the Atlantic but it is by no means certain that they will bring about the multidisciplinary approach to providing patients with treatment options which he and others advocate. It also remains to be seen whether the evidence base when it is eventually acquired will vindicate the present increasing dominance of PCI over CABG. PMID- 16719997 TI - Abdominal radiograph requesting in the setting of acute abdominal pain: temporal trends and appropriateness of requesting. AB - INTRODUCTION: The biannual turnover of house surgeons has long been dreaded by paramedical staff because of fears of increased workloads generated by 'untrained' junior doctors. The aim of this study was to address this issue by examining both the quantity and quality of requests made for emergency abdominal radiographs made by 'experienced' house surgeons during the month of July and by the 'novices' during August. PATIENTS AND METHODS: All adult patients undergoing abdominal radiography (AXR) following admission as emergencies via the surgical directorate with abdominal signs were identified prospectively. The reports of the AXRs were reviewed to determine the total number of requests and the number of positive findings for the two groups. In addition, the hand-written request forms were recovered to determine the suitability of the requests according to nationally-accepted guidelines produced by the Royal College of Radiologists (RCR). RESULTS: During the study period, a total of 252 radiographs were performed consisting of 98 in July and 154 in August. The number of unreported films in each month were similar at 11 (11.2%) and 16 (10.4%), respectively, leaving 87 reported radiographs in July and 138 in August. There was no difference in the number of radiographs with positive findings (excluding degenerative spinal disease) for July (n = 19; 22%) and August (n = 33; 24%). Of the 225 reported films, RCR guidelines were followed in only 73 (32%) of 225 cases. When guidelines were adhered to, positive findings were identified in 56 (76.7%) of 73 cases whereas when guidelines were not followed positive findings were seen in only 13/139 (8.9%) of AXRs. CONCLUSIONS: We have demonstrated that the popular myth of the 'August syndrome' is unsubstantiated at least using the surrogate marker of abdominal radiograph requests. The worrying finding of a high number of unacceptable indications for the performance of abdominal radiographs deserves urgent attention both in terms of its financial implications and with regards reducing radiation exposure. A programme of education is proposed to emphasise the RCR guidelines with re-audit to assess adherence to the guidelines. PMID- 16719998 TI - A prospective audit of early pain and patient satisfaction following out-patient band ligation of haemorrhoids. AB - INTRODUCTION: Information regarding early morbidity, pain and patient satisfaction following band ligation of haemorrhoids is limited. This is the first report to address these issues specifically. PATIENTS AND METHODS: A total of 183 patients underwent the procedure over a 10-month period. Prospective data were collected using a detailed structured questionnaire regarding symptoms, analgesia requirements and patient satisfaction in the following week. RESULTS: The response rate was 74% (135/183). Pain scores were highest 4 h following the procedure. At 1 week, 75% of patients were pain-free, with 9 (7%) still experiencing moderate-to-severe pain. About 65% required oral analgesia, most frequently on the day of procedure. Rectal bleeding occurred in 86 patients (65%) on the day after banding, persisting in 32 (24%) at 1 week. Vaso-vagal symptoms occurred in 41 patients (30%) and were commonest at the time of banding. Eighty patients (59%) were satisfied with their experience and would undergo the procedure again. Patients requiring oral analgesia and those experiencing bleeding or vaso-vagal symptoms were significantly less likely to be satisfied with the procedure. Only 57% of the patients surveyed would recommend the procedure to a friend. CONCLUSIONS: Data from this large cohort of patients suggest that discomfort and bleeding may persist for a week or more following banding of haemorrhoids. Patients should be aware of this in order to make an informed decision as to whether to undergo the procedure, and surgeons should investigate ways of reducing it. Patient satisfaction may be further improved by more accurate counselling regarding the incidence of specific complications. PMID- 16719999 TI - The management of aspirin in transurethral prostatectomy: current practice in the UK. AB - INTRODUCTION: Stopping aspirin prior to transurethral prostatectomy (TURP) may minimise peri-operative blood loss, but it may also increase the risk of a significant cardiovascular event. There are no guidelines on the management of aspirin in TURP. This study sought to determine the variation in the peri operative management of transurethral prostatectomy (TURP) patients that are taking aspirin. MATERIALS AND METHODS: A questionnaire was sent to 444 consultant urologists in the UK from a list obtained from the British Association of Urologists. This resulted in 290 anonymous replies (65%), of which 287 were suitable for analysis. RESULTS: Of these 287 urologists, 178 (62%) ask patients to stop aspirin prior to TURP. Aspirin is stopped 9.8 days (median, 10 days; range, 2-30 days) prior to surgery, and recommenced 8.8 days (median, 7 days; range, 1-42 days) after surgery. In those that stop aspirin, 62% will stop aspirin in all patients, regardless of the indication, and 40% will cancel a TURP if aspirin use has inadvertently continued. Of the 287 urologists, 109 (38%) do not stop aspirin. CONCLUSIONS: There is a wide variation in the management of aspirin in TURP patients in the UK. Aspirin is being stopped in patients at high risk of serious cardiovascular disease, often for longer than necessary. There is a need for multidisciplinary guidelines to reduce variation in practice. PMID- 16720000 TI - PSA testing: are patients aware of what lies ahead? AB - INTRODUCTION: Screening for prostate cancer with serum prostate specific antigen (PSA) remains a controversial topic. The UK NHS Executive has issued extensive guidance stressing the importance of adequate counselling prior to performing this test. This study aims to assess men's knowledge of the PSA test at the time of their referral and their attitude towards screening. PATIENTS AND METHODS: A total of 219 men referred to urology via the 'fast track' prostate cancer service were recruited into the study. Of these, 191 were referred from primary care and 28 from secondary care. All men completed a questionnaire regarding their knowledge and expectation of the test. RESULTS: The response rate for completed questionnaires was 100%. Overall, 91 (41.5%) men were aware that their PSA had been performed prior to referral and only 79 (36%) men understood why the test was being done. Patients referred from secondary care appeared to be better informed. Despite these figures, 175 (80%) men said they would recommend PSA testing to a friend or colleague, and 196 (89%) men said the test should be broadly publicised. CONCLUSIONS: Nearly two-thirds of the men referred to urology with an elevated PSA were unaware that they had even had their PSA done. Information about the limitations of PSA testing and the consequence of a positive test result had been deficient. Informed counselling for the PSA test should form part of the consultation of any physician intending to undertake this test whether for lower urinary tract symptoms or for prostate cancer screening. PMID- 16720001 TI - Improving hand-washing performance - a crossover study of hand-washing in the orthopaedic department. AB - INTRODUCTION: Effective hand-washing is essential for reducing the spread of infection in hospitals. We aimed to evaluate hand-washing performance of hospital personnel and to determine if this could be improved by education. MATERIALS AND METHODS: A total of 55 personnel working in the orthopaedic department were asked to clean their hands using an alcohol gel containing a clear fluorescent substance. They were unaware of the assessment method. Performance was assessed by examining their hands under UV light to identify areas that had been neglected. Subjects could visualise which areas they had missed and were then educated regarding hand-washing technique and retested after 7 days. RESULTS: Of the 55 subjects, 53 completed the study. Individual performance varied widely. Following a simple educational intervention, 49 out of 53 subjects improved from an average of 7.8% to 2.3% area missed (P < 0.001). CONCLUSIONS: We suggest that hand-washing effectiveness needs to be improved and that a simple educational intervention can be effective. PMID- 16720002 TI - Ionising radiation exposure to orthopaedic trainees: the effect of sub-specialty training. AB - INTRODUCTION: We monitored image intensifier use by orthopaedic trainees to assess their exposure to ionising radiation and to investigate the influence of sub-specialty training. MATERIALS AND METHODS: Five different orthopaedic registrars recorded their monthly image intensifier screening times and exposure doses for all cases (trauma and elective), for a combined total of 12 non consecutive months. Radiation exposure was monitored using shoulder and waist film badges worn both by surgeons and radiographers screening their cases. RESULTS: Registrars in spinal sub-specialties were exposed to significantly higher doses per case and cumulative doses per month than non-spinal trainees (P < 0.05), but significantly lower screening times per case (P < 0.05). There were no significant differences in cumulative screening times per month (P > 0.05). Regression analysis for all surgeons showed a significant relationship between shoulder film badge reading and cumulative dose exposed per month (P < 0.05), but not for cumulative screening time. Shoulder film badge recordings were significantly higher for spinal compared with non-spinal registrars (P < 0.05), although all badges were below the level for radiation reporting. Only one radiographer badge recorded a dose above threshold. CONCLUSIONS: Whilst the long term effects of sub-reporting doses of radiation are not fully understood, we consider that this study demonstrates that trainees should not be complacent in accepting inadequate radiation protection. The higher doses encountered with spinal imaging means that sub-specialty trainees should be alerted to the risk of their increased exposure. The principle of minimising radiation exposure must be maintained by all trainees at all times. PMID- 16720003 TI - Above-knee versus below-knee stockings in total knee arthroplasty. AB - INTRODUCTION: Graduated compression stockings are frequently used following arthroplasty surgery for deep vein thrombosis (DVT) prophylaxis. There are often strongly held beliefs regarding whether below-knee or above-knee stockings should be used in total knee joint replacement. Many surgeons argue that below-knee stockings are more likely to induce wound complications because of increased swelling above the stocking and the elastic causing localised pressure on the wound. PATIENTS AND METHODS: This study is a prospective comparison of above and below-knee stockings in patients undergoing total knee joint replacement. A total of 50 patients were recruited and studied - 23 patients in the above-knee stocking group and 24 in the below-knee group. Swelling around the knee was measured over the course of an in-patient stay, together with wound complication rates and patients' preference. RESULTS: No statistical difference was found between the groups and no difference in wound complication rates was determined. CONCLUSIONS: Below-knee stockings are safe following total knee replacement and are preferred by patients. PMID- 16720004 TI - Mammography in symptomatic women attending a rapid diagnosis breast clinic: a prospective study. AB - INTRODUCTION: We determined whether it is safe to avoid mammograms in a group of symptomatic women with a non-suspicious history and clinical examination. PATIENTS AND METHODS: Symptomatic women aged 35 years or over newly referred to a rapid-diagnosis breast clinic underwent mammography on arrival in the clinic. A breast radiologist reported on the mammograms. An experienced clinician who was unaware of the mammogram findings examined patients and decided whether a mammogram was indicated or not. If not, a management plan was formulated. Mammogram findings were then provided to the clinician and any change to the original management plan as a result of mammography was recorded. RESULTS: In two thirds (67%) of 218 patients, the clinician felt a mammogram was indicated. Half (46%) of these mammograms showed an abnormality; of these abnormal mammograms, 41% were breast cancer. Among the third (n = 71) of mammograms felt not to be indicated, 3 showed abnormalities of which 2 were breast cancer. One cancer was not suspected clinically or mammographically but was diagnosed on cyto/histopathological assessment. CONCLUSIONS: A significant proportion of patients attending a symptomatic breast clinic have a non-suspicious history and normal clinical findings on examination. However, even in this group avoiding mammograms risks missing clinically occult breast cancers. It would appear sensible to offer mammograms to all symptomatic women over 35 years of age. PMID- 16720005 TI - How should a candidate assess varicose veins in the MRCS clinical examination? A vascular viewpoint. AB - INTRODUCTION: Varicose veins are a common problem and, therefore, regularly feature in the vascular bay of the MRCS clinical examination. Candidates are still being instructed to perform tests in the examination that are considered by many to be obsolete and inaccurate. Using the current cohort of vascular examiners, we aim to clarify which tests a candidate should be performing when assessing varicose veins. We also aim to assess basic surgical trainees' experience in the use of hand-held Doppler (HHD). MATERIALS AND METHODS: Postal questionnaires were sent to all English College Court examiners with a declared vascular interest to gain their opinion on what tests should be used in the vascular bay to assess primary varicose veins. E-mail questionnaires were also sent to basic surgical trainees to assess their experience in the use of hand held Doppler to assess varicose veins. RESULTS: There was a 100% response rate from the examiners with 93%, 86% and 79% feeling that clinical examination, HHD examination of the SFJ and HHD examination of the SPJ, respectively, should form part of the examination of primary varicose veins in the vascular bay. Only 50% indicated the Trendelenburg test and cough impulse and 57% believed the tap test should form part of the examination of varicose veins. Of the BSTs, 53% believed they could examine varicose veins with HHD. Of the BSTs who could use HHD, 74% had held a vascular SHO post. DISCUSSION: Published data and opinion show many consultant surgeons have totally abandoned the use of the Trendelenberg, cough, tap and Perthes tests and support the opinion that HHD increases the accuracy of the examination of varicose veins. This study shows the opinions of the examiners supports the evidence-based recommendations that, in the light of easily accessible HHD, the older tests are now outdated. The majority of BSTs who were able to use HHD had held a vascular SHO post (74%) but otherwise it was unlikely that the BST would be comfortable with this skill. CONCLUSIONS: The Brodie Trendelenburg (tourniquet) test, cough impulse and tap test are outdated but candidates should be aware of the principles and failings behind them. In the MRCS clinical examination, candidates should examine varicose veins by means of clinical examination and HHD as this is now accepted standard practice. To aid candidate education, the HHD technique should replace traditional clinical tests which continue to be taught in medical school and remain within the classical surgical text books. PMID- 16720006 TI - Audit of doctors' knowledge of major incident policies. AB - INTRODUCTION: The aim of this study was to audit doctors' knowledge of their hospital's major incident policy. MATERIALS AND METHODS: Pre- and post intervention interviews were conducted with doctors selected at random in a central London teaching hospital on where to report to in a major incident. Doctors working in hospitals with accident and emergency departments within the M25 motorway were asked if they had read or received training on their hospital's major incident policy. RESULTS: Pre-intervention, 4.4% of doctors knew where to report to in a major incident. A 1-sided information sheet on the major incident policy was distributed to doctors and posted in areas frequented by doctors. Following this intervention, 78% of doctors knew where to report to in a major incident. Doctors in only 2 out of 38 hospitals with accident and emergency departments within the M25 corridor had read or received training on their major incident policy. CONCLUSIONS: More needs to be done by hospitals to ensure that doctors have a better awareness of their hospital major incident policy. PMID- 16720009 TI - Comment on: Intervention to decompress the upper tracts in patients with established pelvic malignancies. PMID- 16720010 TI - Comment on: The use of the laryngeal mask in surgical tracheostomy. PMID- 16720011 TI - Comment on: Alternative swabs. PMID- 16720012 TI - Comment on: Percutaneous tracheostomy. PMID- 16720013 TI - Antihyperlipidemic and antidiabetic effects of umbelliferone in streptozotocin diabetic rats. AB - The aim of the study was to evaluate blood glucose and lipid lowering effects of Umbelliferone (UMB) in streptozotocin (STZ) diabetic rats. Male albino Wistar rats (180 to 200 g) were induced diabetes by administration of STZ (40 mg/kg) intraperitonially. Normal and diabetic rats were treated with UMB in 10 percent dimethyl sulfoxide (DMSO) for 45 days. Diabetic rats had increased plasma glucose and decreased insulin, total proteins (TP), and albumin in addition to decreased food intake and body weight. Elevation in total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), very low density lipoprotein cholesterol (VLDL C), triglycerides (TG), free fatty acids (FFA), and phospholipids (PL), and reduction in high density lipoprotein cholesterol (HDL-C) in the plasma were observed. Liver and kidney tissues of diabetic rats had elevation in the levels of TC, TG, FFA, and PL. Treatment with UMB decreased plasma glucose and increased insulin, TP, and albumin apart from food intake and body weight. In UMB-treated diabetic rats, plasma and tissue TC, TG, PL and FFA, and plasma LDL-C, VLDL-C, and HDL-C reversed to near normal. Thus, reduction of blood glucose and lipid profiles indicates that UMB has antidiabetic and antihyperlipidemic effects in diabetic rats. PMID- 16720014 TI - SUMO: a ubiquitin-like protein modifier. AB - SUMO (Small Ubiquitin-related Modifier) is a small protein that covalently attaches to a lysine residue of another protein in a reversible fashion. SUMO attachment to its substrate proteins causes changes in the localization, activity, or binding partners of the substrate. SUMO has been shown to play a role in a multitude of processes; these include chromosome segregation, cell cycle progression, and DNA damage recovery. Defects in the SUMO pathway have been demonstrated to affect tumorigenesis and the inflammatory response as well as other human diseases. PMID- 16720015 TI - The molecular mechanisms of classic Hodgkin's lymphoma. AB - Classic Hodgkin's lymphoma is characterized by the appearance of giant abnormal cells called Hodgkin and Reed-Sternberg (HRS) cells. HRS cells arise from germinal center B lymphocytes and in about 50 percent of patients, are infected with Epstein-Barr Virus. In addition, HRS cells show constitutive NF-kappaB activation and are resistant to apoptosis. This paper reviews several recent studies that for the first time implicate specific molecules in the pathogenesis of classic Hodgkin's lymphoma. Targeting these molecules could lead to the development of novel therapies for this disease. PMID- 16720016 TI - Management of disorders of the posterior pelvic floor. AB - INTRODUCTION: Constipation is a relatively common problem affecting 15 percent of adults in the Western world, and over half of these cases are related to pelvic floor disorders. This article reviews the clinical presentation and diagnostic approach to posterior pelvic floor disorders, including how to image and treat them. METHODS: A Pubmed search using keywords "rectal prolapse," "rectocele," "perineal hernia," and "anismus" was performed, and bibliographies of the revealed articles were cross-referenced to obtain a representative cross-section of the literature, both investigational studies and reviews, that are currently available on posterior pelvic floor disorders. DISCUSSION: Pelvic floor disorders can occur with or without concomitant physical anatomical defects, and there are a number of imaging modalities available to detect such abnormalities in order to decide on the appropriate course of treatment. Depending on the nature of the disorder, operative or non-operative therapy may be indicated. CONCLUSION: Correctly diagnosing pelvic floor disorders can be complex and challenging, and the various imaging modalities as well as clinical history and exam must be considered together in order to arrive at a diagnosis. PMID- 16720017 TI - Peripherin- and CGRP-immunoreactive nerve fibers in rat molars have different locations and developmental timing. AB - Developing rat molars gain mature sensitivity to electric stimulation at 4-5 weeks after eruption, but the related mechanisms are incompletely understood. Preliminary studies showed weak co-localization of calcitonin gene-related peptide (CGRP) immunoreactivity (IR) with peripherin (PER) or neurofilament protein (NF) in rat molar nerve fibers, while the latter two co-localized extensively. OBJECTIVE: Our goal was to compare timing and location of PER-IR and CGRP-IR innervation in rat first molars during tooth maturation. METHODS: We used single and double immunocytochemistry to study molars of rats aged 10 days to 1 year. Neural patterns were compared with odontoblast maturation stages, dentinogenesis, formation of cell-free and cell-rich zones, and root closure. RESULTS: Spatial and temporal patterns showed that most CGRP-IR and PER-IR have different terminal domains in teeth. PER-IR fibers were well established among immature odontoblasts prior to tooth eruption, but CGRP-IR fibers were absent. Two weeks after eruption of first molars, many CGRP-IR beaded fibers entered dentin, the larger PER-IR fibers began shifting away from odontoblasts towards the pulp, and the symmetrical PER-IR pulpal pattern was being established. The CGRP-IR fibers continued to increase their asymmetric dentinal innervation until root growth was completed, during which time odontoblasts matured, the cell-free and cell-rich zones appeared, and roots closed. CONCLUSIONS: Sensory maturation of rat molars coincides with closed root apices, extensive innervation of dentin by CGRP-IR nerve fibers, and the appearance of the mature avascular odontoblast layer next to cell-free and cell-rich zones in the pulp horns. PMID- 16720018 TI - 3'-Azido-3'-deoxythymidine (AZT) is a competitive inhibitor of thymidine phosphorylation in isolated rat heart and liver mitochondria. AB - Long-term use of 3'-azido-3'-deoxythymidine (AZT) is associated with various tissue toxicities, including hepatotoxicity and cardiomyopathy, and with mitochondrial DNA depletion. AZT-5'-triphosphate (AZTTP) is a known inhibitor of the mitochondrial DNA polymerase gamma and has been targeted as the source of the mitochondrial DNA depletion. However, in previous work from this laboratory with isolated rat heart and liver mitochondria, AZT itself was shown to be a more potent inhibitor of thymidine phosphorylation (IC50 of 7.0+/-1.0 microM AZT in heart mitochondria and of 14.4+/-2.6 microM AZT in liver mitochondria) than AZTTP is of polymerase gamma (IC50 of >100 microM AZTTP), suggesting that depletion of mitochondrial stores of TTP may limit replication and could be the cause of the mitochondrial DNA depletion observed in tissues affected by AZT toxicity. The purpose of this work is to characterize the nature of AZT inhibition of thymidine phosphorylation in isolated rat heart and rat liver mitochondria. In both of these tissues, AZT was found to be a competitive inhibitor of the phosphorylation of thymidine to TMP, catalyzed by thymidine kinase 2. The inhibition constant (Ki) for heart mitochondria is 10.6+/-4.5 microM AZT, and for liver mitochondria Ki is 14.0+/-2.5 microM AZT. Since AZT is functioning as a competitive inhibitor, increasing thymidine concentrations may be one mechanism to overcome the inhibition and decrease AZT-related toxicity in these tissues. PMID- 16720020 TI - Polychlorinated biphenyl exposure delays metamorphosis and alters thyroid hormone system gene expression in developing Xenopus laevis. AB - Polychlorinated biphenyls (PCBs) are ubiquitous environmental contaminants that disrupt thyroid hormone (TH) system function in numerous species. Previous studies have shown delayed metamorphosis in developing Xenopus laevis frogs exposed to PCBs, but the underlying molecular mechanisms have not been thoroughly investigated. In this research, developing X. laevis tadpoles were exposed to environmentally relevant concentrations (5, 50ppb) of Aroclor 1254 (A1254), a PCB mixture, dissolved in water and 0.25% dimethyl sulfoxide. Quantitative real-time reverse transcriptase polymerase chain reaction was used to measure expression of several TH system genes, other genes that regulate growth and development, and a xenobiotic response gene. Exposure to 50ppb A1254 significantly delayed metamorphosis and significantly altered gene expression of three thyroid system genes: transthyretin and types II and III deiodinase. Since all three genes regulate the amount of available, biologically active TH, PCB-induced changes in the expression of these genes may underlie alterations in metamorphic timing. PMID- 16720019 TI - Evaluation of nitroreductase and acetyltransferase participation in N nitrosodiethylamine genotoxicity. AB - N-Nitroso compounds, such as N-nitrosodiethylamine (NDEA), are a versatile group of chemical carcinogens, being suspected to be involved in gastrointestinal tumors in humans. The intestinal microflora can modify a wide range of environmental chemicals either directly or in the course of enterohepatic circulation. Nitroreductases from bacteria seem to have a wide spectrum of substrates, as observed by the reduction of several nitroaromatic compounds, but their capacity to metabolize N-nitroso compounds has not been described. To elucidate the participation of nitroreductase or acetyltransferase enzymes in the mutagenic activity of NDEA, the bacterial (reverse) mutation test was carried out with the strains YG1021 (nitroreductase overexpression), YG1024 (acetyltransferase overexpression), TA98NR (nitroreductase deficient), and TA98DNP6 (acetyltrasferase deficient), and YG1041, which overexpresses both enzymes. The presence of high levels of acetyltransferase may generate toxic compounds that must be eliminated by cellular processes or can lead to cell death, and consequently decrease the mutagenic effect, as can be observed by the comparison of strain TA98DNP6 with the strains TA98 and YG1024. The slope curves for TA98 strain were 0.66 rev/microM (R(2) = 0.51) and 52.8 rev/microM (R(2) = 0.88), in the absence and presence of S9 mix, respectively. For YG1024 strain, the slope curve, in the presence of S9 mix was 6897 rev/microM (R(2) = 0.78). Our data suggest that N-nitroso compounds need to be initially metabolized by enzymes such as cytochromes P450 to induce mutagenicity. Nitroreductase stimulates toxicity, while acetyltransferase stimulates mutagenicity, and nitroreductase can neutralize the mechanism of mutagenicity generating innoccuos compounds, probably by acting on the product generated after NDEA activation. PMID- 16720021 TI - Different expression of angiogenesis-related factors between human cultivated corneal and oral epithelial sheets. AB - We developed a cultivated oral mucosal epithelial sheet (COE) transplantation system to address severe human ocular surface disorders. Unlike the cultivated corneal epithelial sheet (CCE), the COE induces mild superficial peripheral neovascularization although central clarity is maintained. To evaluate the characteristic differences between CCE and COE regarding to angiogenesis, we examined the expression of angiogenesis-related factors in CCE and COE. Using samples of CCE and COE, we immunohistochemically determined protein expression of the angiogenesis related factors: Thrombospondin-1 (TSP-1), pigment epithelium derived factor (PEDF), endostatin, angiostatin, vascular endothelial growth factor (VEGF), Fms-like tyrosine kinase 1 (Flt-1), kinase insert domain receptor (KDR), and basic fibroblast growth factor (bFGF). We used Western blot analysis to confirm the factors that were immunohistochemically different in CCE and COE. The immunohistochemical staining intensity of TSP-1 was higher in CCE than COE and by Western blot analysis the expression of TSP-1 was significantly higher in CCE than COE (P<0.05). PEDF and endostatin stained moderately stronger in CCE than COE. Immunohistochemically there was no obvious difference between CCE and COE with respect to angiostatin, VEGF, Flt-1, KDR, and bFGF. In comparison with CCE, COE showed decreased expression of anti-angiogenic factors particularly TSP 1. This different expression may relate to the superficial peripheral neovascularization encountered after COE transplantation. PMID- 16720022 TI - Focus on molecules: bestrophin (best-1). PMID- 16720024 TI - How well does the metabolic syndrome defined by five definitions predict incident diabetes and incident coronary heart disease in a Chinese population? AB - We evaluate the ability of the metabolic syndrome (MetS) defined by five definitions for predicting both incident CHD and diabetes combined, diabetes alone, and CHD alone in a Chinese population. The screening survey for type 2 diabetes was conducted in 1994. A follow-up study of 541 high-risk non-diabetic individuals who were free of CHD at baseline was carried out in 1999 in Beijing area. The MetS was defined by the World Health Organization (WHO), European Group for the Study of Insulin Resistance (EGIR), American College of Endocrinology (ACE), the International Diabetes Federation (IDF), and the National Cholesterol Education Program and the American Heart Association (AHA) (updated NCEP) criteria. From a multiple logistic regression adjusting for age, sex, education, occupation, smoking, family history of diabetes, and total cholesterol, the relative risk of the ACE-defined MetS for incident diabetes alone (67 cases) was 2.29 (95% CI, 1.20-4.34). The MetS defined by the five definitions was associated with a 1.8-3.9 times increased risk for both incident CHD and diabetes combined (59 cases), and with a 1.9-3.0 times for total incident diabetes (126 cases). None of the five definitions predicted either incident CHD alone (177 cases) or total incident CHD (236 cases). In conclusion, the MetS defined by the current definitions appears to be more effective at predicting incident diabetes. PMID- 16720025 TI - Tackling matrix effects during development of a liquid chromatographic electrospray ionisation tandem mass spectrometric analysis of nine basic pharmaceuticals in aqueous environmental samples. AB - When developing an LC-MS/MS-method matrix effects are a major issue. The effect of co-eluting compounds arising from the matrix can result in signal enhancement or suppression. During method development much attention should be paid to diminish matrix effects as much as possible. The present work evaluates matrix effects from aqueous environmental samples in the simultaneous analysis of a group of nine specific pharmaceuticals with LC-ESI/MS/MS: flubendazole, propiconazole, pipamperone, cinnarizine, ketoconazole, miconazole, rabeprazole, itraconazole and domperidone. Solutions to diminish signal suppression were examined: optimisation of the sample preparation, decrease of the flow rate, and the use of appropriate internal standards. Several SPE-stationary phases were tested in view of retention of the analytes: Oasis HLB, C8, Phenyl, Strata X polymer RP sorbent and Strata X-polymeric SCX/RP sorbent. Oasis HLB showed the best retention for all analytes. The Oasis HLB SPE-method was optimised, but analyses showed high matrix suppression. Therefore, a second SPE-method, on a phenyl stationary phase (the second best option), was also optimised. A comparison of the matrix effect was made between the two procedures: the phenyl method was less subject to matrix effects, however, the average matrix effect (ME%) of 46% indicated that matrix effects where still present. Several optimisation options for the phenyl-method were evaluated: addition of a ferric nitrate solution before extraction, application of an alkaline wash step, and use of a second SPE-cartridge, either a NH2-column or a florisil column. A more efficient sample clean-up was achieved by applying the extract after extraction on the phenyl column and after dilution with chloroform, onto a NH2-column (average ME%: 53%). In addition, applying a post-column split (1:5), further reduced matrix effects (average ME%: 65%). Labelled internal standards are the best way to tackle matrix effects, but no such internal standards were commercially available for the analytes of interest. The thorough search and application of four internal standards (structural analogues) was beneficial and compensates the matrix effect partially (average ME%: 83%). In an attempt to reduce the analysis time Speedisk phenyl columns were applied. Under these conditions matrix effects decreased even more while recoveries were between 91 and 109%. Different kinds of surface water samples were analyzed, and different matrix effects were observed. For this reason, standard addition will be used to perform quantitative analysis. PMID- 16720023 TI - Biomechanics and wound healing in the cornea. AB - The biomechanical and wound healing properties of the cornea undermine the predictability and stability of refractive surgery and contribute to discrepancies between attempted and achieved visual outcomes after LASIK, surface ablation and other keratorefractive procedures. Furthermore, patients predisposed to biomechanical failure or abnormal wound healing can experience serious complications such as keratectasia or clinically significant corneal haze, and more effective means for the identification of such patients prior to surgery are needed. In this review, we describe the cornea as a complex structural composite material with pronounced anisotropy and heterogeneity, summarize current understanding of major biomechanical and reparative pathways that contribute to the corneal response to laser vision correction, and review the role of these processes in ectasia, intraocular pressure measurement artifact, diffuse lamellar keratitis (DLK) and corneal haze. The current understanding of differences in the corneal response after photorefractive keratectomy (PRK), LASIK and femtosecond assisted LASIK are reviewed. Surgical and disease models that integrate corneal geometric data, substructural anatomy, elastic and viscoelastic material properties and wound healing behavior have the potential to improve clinical outcomes and minimize complications but depend on the identification of preoperative predictors of biomechanical and wound healing responses in individual patients. PMID- 16720026 TI - Application of reverse-flow micellar electrokinetic chromatography for the simultaneous determination of flavonols and terpene trilactones in Ginkgo biloba dosage forms. AB - A reverse-flow micellar electrokinetic chromatographic (RF-MEKC) method was developed for the simultaneous qualitative determination of 10 components consisting of the flavonol glycosides, rutin and quercitrin, the flavonol aglycones, isorhamnetin, kaempferol and quercetin, the terpene trilactones, ginkgolides A, B, C and J and the sesquiterpene, bilobalide. This method was used to fingerprint Ginkgo biloba solid oral dosage forms and validated for the quantitation of the marker compounds, rutin and quercetin in some commercial products. In addition to the usual variables, the influence of some essential background electrolyte (BGE) components such as sodium dodecyl sulphate (SDS) and -cyclodextrin concentrations were investigated. A polyimide fused-silica square capillary column (75 microm I.D. x 360 microm O.D.) with a total length of 60.0 cm and effective length of 45.0 cm was used for the separation. The final BGE consisted of 20 mM phosphoric acid, 40 mM SDS and 12 mM -cyclodextrin (pH 2.2) using reverse polarity with a voltage of -17.5 kV. Samples were injected electrokinetically at -5 kV for 3 s for the qualitative analysis and hydrodynamically at 20 mbar for 0.6 s for the quantitative assay. The total run time was 22 min and the limits of detection were 3.13 microg/ml and 1.88 microg/ml for rutin and quercetin, respectively. Fingerprint profiles of the solid oral dosage forms and the results of the quantitative analysis indicated that there were major discrepancies in the marker content between products and illustrates the value of this method for use as a procedure to assess product quality of commercially available Ginkgo biloba products. PMID- 16720027 TI - Fast, comprehensive online two-dimensional high performance liquid chromatography through the use of high temperature ultra-fast gradient elution reversed-phase liquid chromatography. AB - A new approach to high speed, comprehensive online dual gradient elution 2DLC (LCxLC) based on the use of ultra-fast, high temperature gradient elution reversed phase chromatography is described. Entirely conventional gradient elution instrumentation and columns are assembled in a system which develops a total peak capacity of about 900 in 25 min; this is equivalent to roughly one peak/2 s. Each second dimension gradient is done in a cycle time of 21 s and the peak retention times measured for a set of twenty six indole-3-acetic acid (IAA) derivatives are reproducible to 0.2 s. Each peak eluting from the first dimension column is sampled at least twice across its width, as the corresponding peaks on the second dimension column appear in two or three consecutive second dimension chromatograms, clearly indicating that there is little loss in the resolution gained in the first dimension separation. Application to the separation of the low molecular weight components of wild-type and mutant maize seedlings indicates the presence of about 100 peaks on a timescale of 25 min. Compelling illustrations of the analytical potential of fast, high temperature 2DLC are evident in the clear presence of nine distinct peaks in a single second dimension chromatogram from a single quite narrow first dimension peak, and the great power of 2DLC to solve the "analytic dynamic range" problem inherent in the measurement of small peaks that are neighbors to a gigantic peak. PMID- 16720028 TI - Open tubular capillary electrochromatography migration behavior of enkephalins in etched chemically modified fused silica capillaries. AB - Fused silica capillaries for use in electrophoretic analyses are etched with ammonium bifluoride in the presence of a second inorganic salt (CuCl(2), CrCl(3), NaNO(3), or (NH(4))(2)CO(3)). The effects of the presence of these inorganic components in the surface matrix on the electromigration behavior of enkephalins are evaluated. Resolution, efficiency and peak shape are used to compare the various columns. In some cases the etched surface is then modified by the addition of an octadecyl moiety using a silanization/hydrosilation procedure. The surface properties of the etched capillaries can also be evaluated by electroosmotic flow measurements. RSDs of migration times under identical experimental conditions were <1%. PMID- 16720029 TI - Human monocytoid cells as a model to study Toll-like receptor-mediated activation. AB - THP-1 2A9, a subclone of the monocytoid cell line THP-1 and known to be exquisitely sensitive to LPS, was tested for TNF production following triggering by excess doses of TLR ligands. TLR2, TLR4 and TLR5 agonists, but neither TLR3 nor TLR9 agonists, induced TNF production. When used at lower concentrations, priming by calcitriol strongly influenced the sensitivity of cells to LPS and different TLR2 triggers (lipoteichoic acid (LTA), trispalmitoyl-cysteyl-seryl lysyl-lysyl-lysyl-lysine (Pam3Cys) and peptidoglycan (PGN)). Priming by calcitriol failed to modulate TLR2 and TLR4 mRNA and cell surface expression of these receptors. TNF signals elicited by TLR2 agonists were blocked by the TLR specific antibody 2392. CD14-specific antibodies showed variable effects. CD14 specific antibodies inhibited TNF induction by LTA. High concentrations partially inhibited TNF induction by Pam3Cys. The same antibodies failed to inhibit TNF induction by PGN. Thus, THP-1 2A9 cells respond by TNF production to some, but not all TLR agonists, and the wide variety of putative TLR2 agonists interact to variable degrees also with other cell-surface-expressed binding sites such as CD14. THP-1 2A9 cells might provide a model by which to investigate in more detail the interaction of pathogen-associated molecular patterns and monocytoid cell-surface-expressed pattern recognition receptors. PMID- 16720030 TI - PDGF-BB enhances monocyte chemoattractant protein-1 mRNA stability in smooth muscle cells by downregulating ribonuclease activity. AB - Platelet-derived growth factor (PDGF) has protean manifestations, including the regulation of growth and migration, in many cell types. We have previously reported that PDGF-BB induces the accumulation of monocyte chemoattractant protein (MCP)-1 mRNA in smooth muscle cells (SMC), in large part due to an increase in mRNA stability. To elucidate the mechanism by which PDGF-BB stabilizes MCP-1 mRNA, we have employed in vitro RNA gel mobility shift and decay assays. Cytoplasmic extracts from PDGF-BB-treated SMC increased the half-life of in vitro transcribed MCP-1 mRNA from approximately 45 min to >2 h. PDGF-BB inhibitable degradation was not dependent on specific regions of the MCP-1 mRNA and was equally effective on a variety of in vitro transcribed mRNAs. Angiotensin II had a similar effect on MCP-1 mRNA stability, whereas tumor necrosis factor alpha and basic fibroblast growth factor did not. The PDGF-BB-inhibitable RNAse activity was active at pH 6.6 and heat stable, but was sensitive to proteinase K. Extracts from PDGF-BB- or angiotensin II-treated cells inhibited the RNAse activity of control extracts, suggesting that the effect of PDGF-BB and angiotensin II are due to activation of a soluble inhibitor of the RNAse. The effect of PDGF-BB was blocked by inhibitors of tyrosine phosphorylation, but not by inhibitors of phosphatidylinositol 3-kinase or mitogen-activated protein kinases. These studies provide new insights into the mechanisms by which PDGF-BB enhances mRNA accumulation. PMID- 16720031 TI - Fusiform type alexia: pure alexia for words in contrast to posterior occipital type pure alexia for letters. AB - OBJECTIVE: To clarify the behavioral differences between patients with pure alexia from different lesions. METHODS: Two patients with pure alexia caused by damage to the fusiform or posterior occipital gyri were given reading and writing tests including kanji (Japanese morphograms) and kana (Japanese phonetic writing). RESULTS: Patient 1 (pure alexia from a fusiform gyrus lesion) had difficulty reading both kanji and kana, with kanji reading more impaired, and imageability and visual complexity effects (imageable or less complex words/characters were read better than nonimageable or more complex words/characters), whereas patient 2 (pure alexia from a posterior occipital gyri lesion) showed selective impairment of kana reading. CONCLUSION: Pure alexia for kanji (and kana; fusiform type) is characterized by impairments of both whole word reading, as represented in kanji reading, and letter identification, and is different from pure alexia for kana (posterior occipital type) in which letter identification is primarily impaired. Thus, fusiform type pure alexia should be designated pure alexia for words, whereas posterior occipital type pure alexia should be designated pure alexia for letters. PMID- 16720032 TI - Quantifying the contribution of actin networks to the elastic strength of fibroblasts. AB - The structural models created to understand the cytoskeletal mechanics of cells in suspension are described here. Suspended cells can be deformed by well-defined surface stresses in an Optical Stretcher [Guck, J., Ananthakrishnan, R., Mahmood, H., Moon, T.J., Cunningham, C.C., Kas, J., 2001. The optical stretcher: a novel laser tool to micromanipulate cells. Biophys. J. 81(2), 767-784], a two-beam optical trap designed for the contact-free deformation of cells. Suspended cells have a well-defined cytoskeleton, displaying a radially symmetric actin cortical network underlying the cell membrane with no actin stress fibers, and microtubules and intermediate filaments in the interior. Based on experimental data using suspended fibroblasts, we create two structural models: a thick shell actin cortex model that describes cell deformation for a localized stress distribution on these cells and a three-layered model that considers the entire cytoskeleton when a broad stress distribution is applied. Applying the models to data, we obtain a (actin) cortical shear moduli G of approximately 220 Pa for normal fibroblasts and approximately 185 Pa for malignantly transformed fibroblasts. Additionally, modeling the cortex as a transiently crosslinked isotropic actin network, we show that actin and its crosslinkers must be co localized into a tight shell to achieve these cortical strengths. The similar moduli values and cortical actin and crosslinker densities but different deformabilities of the normal and cancerous cells suggest that a cell's structural strength is not solely determined by cytoskeletal composition but equally importantly by (actin) cytoskeletal architecture via differing cortical thicknesses. We also find that although the interior structural elements (microtubules, nucleus) contribute to the deformed cell's exact shape via their loose coupling to the cortex, it is the outer actin cortical shell (and its thickness) that mainly determines the cell's structural response. PMID- 16720033 TI - Cariporide inhibits high glucose-mediated adhesion of monocyte-endothelial cell and expression of intercellular adhesion molecule-1. AB - The aim of this study was to examine whether cariporide, a new inhibitor of Na(+)/H(+) exchanger 1 (NHE-1), may inhibit high glucose-induced monocyte endothelial cell adhesion and the expression of intercellular adhesion molecule-1 (ICAM-1). Cultured endothelial cells were incubated with normal glucose control (5.5 mM), cariporide control (5.5 mM glucose plus 10 microM cariporide), hyperosmolarity (5.5 mM glucose plus 16.5 mM mannitol), high glucose (HG, 22 mM), low-concentration cariporide (22 mM glucose plus 0.1 microM cariporide), medium concentration cariporide (22 mM glucose plus 1 muM cariporide), and high concentration cariporide (22 mM glucose plus 10 microM cariporide) for 24 h. Monocytes were isolated from peripheral human blood. Adhered monocytes were quantified by measuring their protein content. ICAM-1 expression and NHE-1 activity was determined with enzyme-linked immunosorbent assay (ELISA) and pH sensitive fluorescent spectrophotometry. Exposure of endothelial cells to HG for 24 h caused an increase of adhesion of monocytes to endothelial cells and an increased expression of ICAM-1. However, these effects were reversed by treatment with cariporide (0.1, 1, 10 microM) in a concentration-dependent manner. Furthermore, cariporide (1 microM) was able to inhibit the activation of NHE-1 induced by HG in endothelial cells. These findings suggest that cariporide might inhibit HG-mediated monocyte-endothelial cell adhesion and expression of ICAM-1 by inhibiting the activation of NHE-1. PMID- 16720034 TI - In situ determination of solidosity profiles during activated sludge electrodewatering. AB - Two non-invasive techniques were evaluated for the on-line measurement of sludge solidosity profiles during both pressure and electrodewatering operations. In a first approach, a radioactive tracer adsorbed onto the sludge solids was monitored by a gamma camera. Although this technique appeared very flexible in use, the lack of resolution highly limited its usefulness for (electro)dewatering experiments. Improvement in gamma camera resolution by the development of new detectors might, however, increase the future applicability of this technique. In a second technique, nuclear magnetic resonance measurements on a specially designed electrodewatering unit were made. Hereby, reliable on-line measurements of the solidosity profiles of activated sludge during electrodewatering could be made, with a resolution of less than 1mm. Thus, the mechanisms of electroosmotic- and pressure-driven cake dewatering could be illustrated. Given the measurement time required for measuring one sludge profile, both techniques appeared mainly suited for slowly varying processes, such as activated sludge expression, and not for fast changing processes, such as the initial phases of sludge filtration. PMID- 16720035 TI - Development of an innovative vertical submerged membrane bioreactor (VSMBR) for simultaneous removal of organic matter and nutrients. AB - A novel vertical submerged membrane bioreactor (VSMBR) composed of anoxic and oxic zones in one reactor was developed in an attempt to reduce the problems concerning effective removal of pollutants from synthetic wastewater including glucose as a sole carbon source as well as membrane fouling. The optimal volume ratio of anoxic zone/oxic zone was found as 0.6. The desirable internal recycle rate and hydraulic retention time (HRT) for effective nutrient removal were 400% and 8h, respectively. Under these conditions, the average removal efficiencies of total nitrogen (T-N) and total phosphorus (T-P) were 75% and 71%, respectively, at the total chemical oxygen demand (T-COD)/T-N ratio of 10. In addition, the VSMBR showed high specific removal rates of nitrogen and phosphorus while the biomass growth yield from the reactor was about 20% of the conventional activated sludge process. PMID- 16720036 TI - Neoplastic transformation of the thyroid gland is accompanied by changes in cellular sialylation. AB - Cancer of the thyroid gland is one of the most common endocrine diseases. Histological evaluation is often complicated by difficulty in distinguishing between benign and malignant lesions. Abnormal glycosylation of cell structures, including changes in sialylation, is a feature of the neoplastic transformation process. The aim of this study was to evaluate associations between neoplastic changes in the thyroid gland and changes in sialylation, with reference to its terminal linkage type. Lectin histochemistry using three sialic acid-binding lectins: Tritrichomonas mobilensis lectin (TML), which recognizes sialic acid without linkage preference; Maackia amurensis leukoagglutinin (MAL), which preferentially binds alpha-2,3-linked sialic acid; and Sambucus nigra agglutinin (SNA), which preferentially binds alpha-2,6-linked sialic acid, were used for detection of sialylated glycoconjugates in 50 human thyroid gland specimens. These included papillary, follicular, oncocytic, medullary and anaplastic carcinomas, follicular adenomas and benign follicular and parenchymatous goiter. The luminal surface of follicular cells in normal thyroid glands, adenomas and goiters showed weak or absent labelling for sialic acid. Malignant transformation of the gland was accompanied by an increase of sialic acid positivity on follicular epithelial cells, especially of alpha-2,3-linked sialic acid. Strong luminal positivity for sialic acid was found in papillary carcinomas, whereas moderate positivity was seen in follicular carcinomas. Inconsistent, weak positivity for sialic acid was documented in medullary and anaplastic carcinomas. Increased membrane sialic acid on thyroid gland cells may be an important diagnostic pathological finding, that could be useful in distinction of malignant from benign thyroid lesions, especially with respect to aspiration cytology diagnostics. PMID- 16720037 TI - Critical evaluation of scientific articles and other sources of information: an introduction to evidence-based veterinary medicine. AB - The purpose of this paper is to briefly review key concepts regarding critical reading of the scientific literature to make informed decisions, in the context of evidence-based veterinary medicine. Key concepts are reviewed, based on the broader experience in human medicine, with adaptations, as indicated, to veterinary medicine. That a paper has been published in a peer-reviewed journal does not guarantee its credibility; guidelines are given regarding the general merit of different kinds of articles, as well as checklists and criteria that can be used to assess a paper. Specific study designs, their merits and limitations, are briefly discussed. Standard numerical indices for assessment of studies involving treatments and for assessments of diagnostic tests are summarized. Criteria for assessing drug trials are presented. The principles of statistical analysis are described, including practical considerations and common errors. Finally, numerous sources of bias are reviewed. PMID- 16720038 TI - Reduced contraction of skin equivalent engineered using cell sheets cultured in 3D matrices. AB - In order to alleviate their extensive contraction, human fibroblast sheets were cultured in combination with three-dimensional matrices (knitted poly(lactic-co glycolic acid) (PLGA) mesh and collagen-hyaluronic acid (CHA) sponge) to form contiguous dermal constructs for tissue engineering a bilayered skin equivalent. The resulting constructs were viable, and supported the development of bilayered skin equivalents which did not contract over the 4-week culture period. When implanted into full-thickness wounds in nude rats, cultured skin equivalents based on PLGA meshes registered a take rate of 100% and showed an extent of wound contraction that was statistically similar to autografts, while wounds grafted with PLGA meshes without cell sheets contracted more than autografts. On the other hand, skin equivalents based on CHA sponges were all sloughed off within 2 weeks of transplantation. In all cell sheet-incorporated specimens, cells from the constructs infiltrated and produced extracellular matrix within the neo dermis, shown by positive human leukocyte antigen and collagen I expression. This technique offers an alternative approach for scaffold-based tissue engineering to produce mechanically stable grafts with matured neo-tissue. PMID- 16720039 TI - In vitro biodegradation of three brushite calcium phosphate cements by a macrophage cell-line. AB - Depending upon local conditions, brushite (CaHPO4 x 2 H2O) cements may be largely resorbed or (following hydrolysis to hydroxyapatite) remain stable in vivo. To determine which factors influence cement resorption, previous studies have investigated the solution-driven degradation of brushite cements in vitro in the absence of any cells. However, the mechanism of cell-mediated biodegradation of the brushite cement is still unknown. The aim of the current study was to observe the cell-mediated biodegradation of brushite cement formulations in vitro. The cements were aged in the presence of a murine cell line (RAW264.7), which had the potential to form osteoclasts in the presence of the receptor for nuclear factor kappa B ligand (RANKL) in vitro, independently of macrophage colony stimulating factor (M-CSF). The cytotoxicity of the cements on RAW264.7 cells and the calcium and phosphate released from materials to the culture media were analysed. Scanning electron microscopy (SEM) and focused ion beam (FIB) microscopy were used to characterise the ultrastructure of the cells. The results showed that the RAW264.7 cell line formed multinucleated TRAP positive osteoclast-like cells, capable of ruffled border formation and lacunar resorption on the brushite calcium phosphate cement in vitro. In the osteoclast-like cell cultures, ultrastructural analysis by SEM revealed phenotypic characteristics of osteoclasts including formation of a sealing zone and ruffled border. Penetration of the surface of the cement, was demonstrated using FIB, and this showed the potential demineralising effect of the cells on the cements. This study has set up a useful model to investigate the cell-mediated cement degradation in vitro. PMID- 16720040 TI - Cartilage regeneration using mesenchymal stem cells and a PLGA gelatin/chondroitin/hyaluronate hybrid scaffold. AB - The study was to produce a novel hybrid poly-(lactic-co-glycolic acid) (PLGA) gelatin/chondroitin/hyaluronate (PLGA-GCH) scaffold and evaluate its potentials in cartilage repair. The porous PLGA-GCH scaffold was developed to mimic the natural extra cellular matrix of cartilage. The differentiated mesenchymal stem cells (MSCs) seeded on PLGA-GCH or PLGA scaffold were incubated in vitro and showed that, compared to PLGA scaffold, the PLGA-GCH scaffold significantly augmented the proliferation of MSCs and GAG synthesis. Then autologous differentiated MSCs/PLGA-GCH was implanted to repair full-thickness cartilage defect in rabbit, while MSCs/PLGA for the contra lateral cartilage defect (n=30). Fifteen additional rabbits without treatment for defects were used as control. Histology observation showed the MSCs/PLGA-GCH repair group had better chondrocyte morphology, integration, continuous subchondral bone, and much thicker newly formed cartilage compared with MSCs/PLGA repair group 12 and 24 weeks postoperatively. There was a significant difference in histological grading score between these two groups, which both showed much better repair than control. The present study implied that the hybrid PLGA-GCH scaffold might serve as a new way to keep the differentiation of MSCs for enhancing cartilage repair. PMID- 16720041 TI - Identification of arginase in human placental villi. AB - l-Arginine is the common substrate for arginase and nitric oxide synthase (NOS). Arginase converts l-arginine to urea and ornithine, which is the principal precursor for production of polyamines required for cell proliferation. Human placenta expresses endothelial NOS (eNOS) in syncytiotrophoblasts, but the expression of arginase has not been fully elucidated. Our aim was to investigate the expression and distribution patterns of arginase-I (A-I) and arginase-II (A II) in human placental villi in the first trimester and at term using immunohistochemistry, RT-PCR and Western blot analysis. The arginase enzyme activity in placental villi was also measured. Immunohistochemistry showed different distribution patterns of the arginase isoforms during gestation: A-I was observed only in cytotrophoblasts, while A-II was observed in both cytotrophoblasts and syncytiotrophoblasts. RT-PCR and Western blot analysis showed expression of A-I and A-II in the first trimester and at term in human placental villi. Expression of A-II and arginase activity was greater in the first trimester than at term. Differentiation of cytotrophoblasts into syncytiotrophoblasts may be associated with l-arginine metabolism through modulation of l-arginine availability for eNOS and A-I. And elevated arginase activity in the early gestational period may be responsible for proliferation of trophoblasts by increasing polyamines production. These results suggest that the l-arginine-ornithine-polyamine and l-arginine-nitric oxide pathways play a role in placental growth and development. PMID- 16720042 TI - Calcium signals and the in vitro migration of chick ciliary ganglion cells. AB - We have studied calcium signals and their role in the migration of neuronal and nonneuronal cells of embryonic chick ciliary ganglion (CG). In vitro, neurons migrate in association with nonneuronal cells to form cellular aggregates. Changes in the modulus of the velocity of the neuron-nonneuronal cell complex were observed in response to treatments that increased or decreased intracellular calcium concentration. In addition, both cell types generated spontaneous calcium activity that was abolished by removal of extracellular calcium. Calcium signals in neurons could be characterized as either spikes or waves. Neuronal spikes were found to be related to action potential generation whereas neuronal waves were due to voltage-independent calcium influx. Nonneuronal cells generated calcium oscillations that were dependent on calcium release from intracellular stores and on voltage-independent calcium influx. Application of thimerosal, a compound that stimulates calcium mobilization from internal stores, increased: (1) the amplitude of spontaneous nonneuronal oscillations; (2) the area of migrating nonneuronal cells; and (3) the velocity of the neuronal-nonneuronal cell complex. We conclude that CG cell migration is a calcium dependent process and that nonneuronal cell calcium oscillations play a key role in the modulation of velocity. PMID- 16720043 TI - The mitochondrial Na+/Ca2+ exchanger plays a key role in the control of cytosolic Ca2+ oscillations. AB - There is increasing evidence that mitochondria play an important role in the control of cytosolic Ca2+ signaling. We show here that the main mitochondrial Ca2+-exit pathway, the mitochondrial Na+/Ca2+ exchanger, controls the pattern of cytosolic Ca2+ oscillations in non-excitable cells. In HeLa cells, the inhibitor of the mitochondrial Na+/Ca2+ exchanger CGP37157 changed the pattern of the oscillations induced by histamine from a high-frequency irregular one to a lower frequency baseline spike type, surprisingly with little changes in the average Ca2+ values of a large cell population. In human fibroblasts, CGP37157 increased the frequency of the baseline oscillations in cells having spontaneous activity and induced the generation of oscillations in cells without spontaneous activity. This effect was dose-dependent, disappeared when the inhibitor was washed out and was not mimicked by mitochondrial depolarization. CGP37157 increased mitochondrial [Ca2+] and ATP production in histamine-stimulated HeLa cells, but the effect on ATP production was only transient. CGP37157 also activated histamine-induced Ca2+ release from the endoplasmic reticulum and increased the size of the cytosolic Ca2+ peak induced by histamine in HeLa cells. Our results suggest that the mitochondrial Na+/Ca2+ exchanger directly modulates inositol 1,4,5-trisphosphate-induced Ca2+ release and in that way controls cytosolic Ca2+ oscillations. PMID- 16720044 TI - Emergency department evaluation of the febrile traveler. AB - The emergency department evaluation of the febrile traveler presents the emergency physician with a set of unique and often challenging circumstances. In addition to evaluating and managing the usual array of community-acquired infections, the clinician must be prepared to diagnose and treat a host of uncommon and potentially life-threatening pathogens. This diseases range from widespread tropical diseases such as malaria to the more exotic and lethal viral hemorrhagic fevers. A thoughtful approach guided by geographic patterns of illness offers a reliable method for determining the most likely sources of fever in the returned traveler as well as a focused diagnostic and treatment strategy. PMID- 16720045 TI - Diminished percentage of CD4+ T-lymphocytes expressing interleukine-2 receptor alpha in chronic brucellosis. AB - OBJECTIVES AND METHODS: Despite the treatment, a considerable proportion of brucellosis patients develop chronic disease, characterized by atypical clinical picture and/or relapses. Th1 cytokines are critical for the clearance of Brucella infection and diminished production of IL-2 in response to PHA, has been described in chronic brucellosis. In order to investigate the role of IL-2R alpha (CD25) in disease outcome, we evaluated the ex vivo and PHA-induced percentage of peripheral CD4+ T-lymphocytes expressing CD25 in 13 acute brucellosis patients (AB), 22 chronic brucellosis patients (CB), 11 "clinically cured" subjects and 15 healthy volunteers (controls). Simultaneously, CD3+, CD4+ and CD8+ T-lymphocytes subsets were measured. RESULTS: The ex vivo percentage of CD4+/CD25+ T-cells was significantly higher in AB patients compared to "clinically cured" subjects (p=0.005) and controls (p=0.006). By contrast, CD4+/CD25+ T-cells were significantly lower in CB patients (p=0.001). T-lymphocytes subsets did not significantly differ between the groups. After PHA stimulation, CD4+/CD25+ T cells remained significantly lower in CB and specifically in the relapsing form of chronic disease compared to AB (p=0.044, 0.023). Additionally, CD8+ T lymphocytes were found to be significantly increased in CB and mainly in the relapsing subgroup of CB patients compared to AB (p=0.044, 0.011). CONCLUSION: Diminished percentage of peripheral CD4+ T-lymphocytes expressing IL-2R alpha is associated with chronic relapsing brucellosis. PMID- 16720046 TI - ELR+ CXC chemokines and their receptors (CXC chemokine receptor 1 and CXC chemokine receptor 2) as new therapeutic targets. AB - ELR+ CXC chemokines, by direct interaction with their cell surface receptors CXC chemokine receptor 1 (CXCR1) and CXC chemokine receptor 2 (CXCR2), are believed to be crucially involved in the direct migration and activation of leukocytes. ELR+ CXC chemokines are supposed to play a key role in several inflammatory diseases and this makes ELR+ CXC chemokines and their receptors attractive therapeutic targets. The first aim of this review is to discuss the potential pathological role of ELR+ CXC chemokines in different pathologies, including ulcerative colitis (UC), ischaemia/reperfusion injury (RI), bronchiolitis obliterans syndrome (BOS) and tumor progression. Moreover, the most recently described inhibitors of ELR+ CXC chemokines and their therapeutic indications will be reviewed. Finally, the mode of action and the potential therapeutical use of reparixin, a new potent and selective inhibitor of CXCR1/2 activity, and its chemical derivatives are also discussed. PMID- 16720047 TI - Homogeneous PCR nucleobase quenching assays to detect four mutations that cause neuronal ceroid lipofuscinosis: T75P and R151X in CLN1, and IVS5-1G>C and R208X in CLN2. AB - The neuronal ceroid lipofuscinoses (NCLs) are a family of autosomal recessive lysosomal storage diseases characterized by progressive epilepsy, dementia and visual loss. The juvenile form of the disease (onset age 4-8 years with visual loss) is usually caused by mutations in the CLN3 gene, but some cases have been shown to be due to specific mutations in the CLN1 or CLN2 genes, which are usually associated with NCL with onset in infancy or late infancy, respectively. The CLN1 mutations T75P and R151X, and the CLN2 mutations R208X and IVS5-1G>C, are found in many NCL patients with a juvenile presentation that is not due to CLN3 mutation. We have developed and validated a set of assays for these mutations using PCR followed by differential melting of a fluorescently labeled oligo probe, on a Roche LightCycler platform. The nucleobase quenching phenomenon was used to detect probe hybridization. The tests were validated using alternate assays: PCR followed by allele specific restriction enzyme digestion for the CLN1 mutations, and PCR followed by sequencing for the CLN2 mutations. The homogeneous PCR method gave 100% concordance of results with the alternate methods. This new assay, combined with a test for the common 1 kbp deletion in the CLN3 gene, provides a set of DNA-based assays suitable for detection of the most common mutations causing NCL with onset in the juvenile age range. PMID- 16720048 TI - New mouse oligodendrocyte precursor (mOP) cells for studies on oligodendrocyte maturation and function. AB - Oligodendrocyte precursor (OP) cells give rise to mature oligodendrocytes (OL), which are necessary for myelination of axons during CNS development and following damage to the myelin sheath that occurs in demyelinating diseases. To facilitate studies designed to understand OP maturation and OL function, we have developed OP cells that can be grown continuously, expanded, and differentiated into mature OLs. Cultures of late passage mOP cells grown in proliferation medium are highly pure early stage oligodendrocyte precursors where > 90% assume a characteristic bipolar morphology. Immunocytochemical analysis using antibodies that recognize progressive stages of OP maturation (A2B5, NG2, GD3 and O4) confirmed that mOP cells have a stable early stage OP cell phenotype. In addition, mOP cells can be induced to differentiate into mature forms of oligodendrocytes in vitro and in vivo, as characterized morphologically by the presence of multiple processes with secondary and tertiary branches, and by immunostaining and quantitative real-time PCR for the mature oligodendrocyte markers MBP, MAG, PLP, and MOBP. Finally, differentiation of mOP cells was accompanied by up-regulation of mRNA encoding Olig2 but not Olig1, which is consistent with previous findings showing that Olig2 is necessary for specification of oligodendrocytes. These new mOP cells should significantly benefit in vitro and in vivo studies on OP maturation and function. PMID- 16720050 TI - Comorbidity of PTSD and depression following myocardial infarction. AB - BACKGROUND: This study examines comorbidity of posttraumatic stress disorder (PTSD) and depression following myocardial infarction (MI). It has two aims: (a) to examine whether this comorbidity is predicted by the objective severity of the MI and the patients' initial appraisal and stress responses; and (b) to determine whether this comorbidity is associated with PTSD symptomatology, depression, physiological adjustment, and psychosocial adjustment seven months post-MI. METHOD: 116 MI patients were examined twice. At Time 1, within a week of the MI, initial appraisal, stress responses, and depressive reactions were assessed, and medical measures were obtained from patients' hospital records. At Time 2, seven months later, PTSD, depression, psychosocial functioning, and physical adjustment were assessed. RESULTS: Seven months post-MI, 16% of the patients were identified with PTSD; 8% with comorbid PTSD and depression; and 14% with high levels of depression without full PTSD. Initial level of depression was associated with comorbidity of PTSD and depression. While initial stress reactions did not predict comorbidity, they did predict depression, with or without comorbid PTSD. Comorbidity of PTSD and depression was associated with higher levels of adjustment difficulties. DISCUSSION: Comorbidity of PTSD and depression has some clinical significance. While initial level of depression predicts subsequent comorbidity of PTSD and depression, early stress reactions do not differentiate between MI patients who suffer from depression, with or without comorbid PTSD. PMID- 16720049 TI - Systematic, standardized and comprehensive neurological phenotyping of inbred mice strains in the German Mouse Clinic. AB - Neurological and psychiatric disorders are among the most common and most serious health problems in developed countries. Transgenic mouse models mimicking human neurological diseases have provided new insights into development and function of the nervous system. One of the prominent goals of the German National Genome Research Network is the understanding of the in vivo function of single genes and the pathophysiological and clinical consequences of respective mutations. The German Mouse Clinic (GMC) offers a high-throughput primary screen of genetically modified mouse models as well as an in-depth analysis in secondary and tertiary screens covering various fields of mouse physiology. Here we describe the phenotyping methods of the Neurological Screen in the GMC, exemplified in the four inbred mouse lines C57BL/6J, C3HeB/FeJ, BALB/cByJ, and 129S2/SvPas. For our primary screen, we generated "standard operating procedures" that were validated between different laboratories. The phenotyping of inbred strains already showed significant differences in various parameters, thus being a prerequisite for the examination of mutant mouse lines. PMID- 16720051 TI - Protein tyrosine kinase and p38 MAP kinase pathways are involved in stimulation of matrix metalloproteinase-9 by TNF-alpha in human monocytes. AB - Matrix metalloproteinase-9 (MMP-9), through its catalytic and non-catalytic activities, plays critical roles in inflammation, tumor invasion and angiogenesis. Human monocytes actively involved in inflammatory and tumoral states secrete proMMP-9 (92kDa). Endogenous TNF-alpha stimulates MMP-9 gene transcription in monocytes through NF-kappaB activation. In this study, we investigated the intracellular signaling pathways underlying TNF-alpha/NF-kappaB dependent expression of MMP-9 in monocytes using chemical inhibitors that specifically inhibit distinct kinase pathways. We confirmed the expression of MMP 9 by reverse transcription chain reaction (RT-PCR), ELISA and gelatin zymography. PGE2/cAMP inhibitor indomethacin, PI-3K inhibitor wortmannin, PKC inhibitor bisindolylmaleimide and PKA inhibitor H-89 did not affect the levels of released MMP-9. In contrast, MMP-9 mRNA and protein expression was down-regulated by p38 MAPK inhibitor SB203580 and protein tyrosine kinase (PTK) inhibitor tyrphostin 25. These inhibitors increased IkappaB-alpha levels, which correlate with decreased NF-kappaB activation. Although SB203580 induced a decrease in TNF-alpha release, addition of exogenous TNF-alpha did not reverse the inhibitory effect of SB203580 toward MMP-9 thus suggesting that SB203580 could modulate down-stream effects of TNF-alpha. In parallel, TIMP-1 levels decreased in the presence of SB203580. Both kinase inhibitors did not influence the maturation pathway of monocytes. Our results indicate that these two inhibitors of p38 MAPK and PTK pathways could be used as combined targets for inhibiting MMP-9 expression in inflamed tissues. PMID- 16720052 TI - The flavonoid ellagic acid from a medicinal herb inhibits host immune tolerance induced by the hepatitis B virus-e antigen. AB - The aim of this study is to characterize the role of ellagic acid, a flavonoid from a medicinal herb which blocks HBeAg secretion in a HBV infected cell line and in HBeAg transgenic mice, in immune tolerance in chronic HBV infection. Using the mouse strain C57ML/6, HBeAg-producing transgenic mice (HBeAg-Tg), under the control of metal ion-inducible promoter were generated. The effect on immune tolerance of HBeAg-Tg and the release of immune tolerance by the inhibitor of HBeAg secretion, ellagic acid, was tested using T/B cell proliferation, HBeAg/HBeAb production, cytotoxic T-lymphocyte (CTL) and cytokine assays. C57ML/6 based HBeAg-producing HBeAg-Tg mice were tolerant to HBeAg at the T and B-cell level, did not produce antibodies to HBeAg in vivo and in vitro, produced minimal levels of cytokines (IL-4 and IFN-gamma) and decreased CTL responses, while feeding mice with ellagic acid (5mg/kg body weight) blocked the immune tolerance caused by HBeAg. Our results suggest that host immune tolerance induced by HBeAg during HBV infection, a viral strategy to guarantee HBV infection, can be overcome by ellagic acid, thus it can be used as a therapeutic for HBV-carriers. PMID- 16720053 TI - Polycyclic peptide and glycopeptide antibiotics and their derivatives as inhibitors of HIV entry. AB - Antiviral activity and other biological properties of two groups of polycyclic peptides are discussed. Antibiotics of the complestatin-kistamycin group have a structural motif similar to that of the peptide core of antibacterial antibiotics of the vancomycin-teicoplanin group though no amino acid component in the chloropeptin-kistamicin antibiotics is identical to an amino acid incorporated in the peptide core of the antibiotics of the vancomycin-teicoplanin group. Chloropeptins and the hydrophobic several derivatives of antibacterial antibiotics are inhibitors of HIV and some other viruses. They interfere with the viral (i.e. HIV) entry process. Chemical modifications of natural glycopeptide antibiotics led to the compounds with antiviral properties whereas antibacterial properties were lost. These glycopeptide aglycons derivatives can be envisaged as potential lead compounds for application as microbicides against sexual HIV transmission. PMID- 16720054 TI - Glucagon-like peptide-1 inhibits LPS-induced IL-1beta production in cultured rat astrocytes. AB - Glia play an important role in neurotoxicity in neurodegenerative diseases. In this study, we investigated the expression of glucagon-like peptide-1 (GLP-1) and its receptor, and the effects of GLP-1 on lipopolysaccharide (LPS)-induced IL 1beta mRNA expression and IL-1beta production in glia. GLP-1-like immunoreactivity was observed in amoeboid microglia, but not ramified microglia or astrocytes. GLP-1 binding and GLP-1 receptor mRNA expression were observed in both astrocytes and microglia. GLP-1-induced morphological changes in microglia from the ramified type to the amoeboid type, suggesting an increase in production and release of endogenous GLP-1. GLP-1 prevented the LPS-induced IL-1beta mRNA expression, which effect was, in turn, inhibited by pretreatment with SQ22536, an adenylate cyclase inhibitor. GLP-1 also increased cAMP concentration and cAMP response element-binding protein phosphorylation in astrocytes. These results suggest that GLP-1 may be a modulator of inflammation in the central nervous system. PMID- 16720055 TI - Different memory types for generating saccades at different stages of learning. AB - It has been proposed that positional memories encoded in different types of reference frame are used for the reaching hand movement in different stages of learning. However, the types of reference frame employed for generating behavior at each stage of learning remain unclear, particularly for saccades. To examine the types of reference frame for target positions, we analyzed the saccade of monkeys performing an oculomotor task. The task required the animal to make a learning-based saccade to one of the eight landmark positions specified by the color of a fixation point. According to the color of the fixation point, the target landmark position was fixed in all experimental blocks (FIX trial) or altered to other landmark positions block by block (ALTER trial). Although the monkeys learned the target landmark position in both the FIX and the ALTER trials, once the landmarks became invisible, the success rate remained high only in the FIX trials. These results suggest that the target position was learned on the basis of the landmark positions in the early stage of learning. However, the memory of the target position in space was formed after sufficient training. When the fixation point was shifted horizontally by 5 degrees and the landmarks were invisible, the saccades in the ALTER trials were made to the normal target landmark position whereas those in the FIX trials were made to the point approximately 5 degrees shifted horizontally from the normal target landmark position. These results suggest that the target position in space was initially represented in the head-centered or world-centered reference frame and then in the eye-centered reference frame. Analysis of saccade end-points indicated that a kinematically similar saccade was generated for each FIX trial. These results showed that memories encoded by different reference frames were formed to generate a saccade while the saccade toward the same target was repeatedly executed. PMID- 16720056 TI - Characterization of Escherichia coli isolated from carcasses of beef cattle during their processing at an abattoir in Brazil. AB - Beef carcass sponge samples collected between March 2003 and August 2005 at an abattoir in Brazil were surveyed for the presence of Shiga toxin-producing Escherichia coli (STEC). Only one carcass among the 80 tested showed a STEC, stx2 encoding gene by PCR amplification. The frequency of carcass contamination by E. coli during processing was tested at three situations, respectively: preevisceration, postevisceration and postprocessing, during the rain and dry seasons. The prevalence of E. coli at the three points was of 30.0%, 70.0%, 27.5% in the rain season and of 22.5%, 55.0%, 17.5% during the dry season, respectively. The E. coli isolates exhibited a high level (45.0%) of multidrug resistance to two or more antimicrobial agents. PMID- 16720057 TI - Regression of cardiac hypertrophy in type 2 diabetes with hypertension by candesartan. AB - This study was designed to compare the effect of candesartan on cardiac left ventricular mass in Japanese patients with that of amlodipine. A total of 40 type 2 diabetic patients with hypertension and left ventricular hypertrophy (LVH) were randomly assigned to receive candesartan (n=20) or amlodipine (n=20). The two treatments when administered for 6 months significantly reduced systolic and diastolic blood pressures (BPs) to a comparable extent. Notably, candesartan significantly reduced left ventricular mass index (LVMI: from 131.5+/-4.5 to 112.1+/-5.9g/m(2), P=0.0009, M+/-S.E.M.), LV posterior wall thickness (PWTd: from 10.3+/-0.3 to 9.1+/-0.3mm, P=0.0052) and interventricular septal thickness (IVSTd: from 10.7+/-0.4 to 9.3+/-0.4mm, P=0.0019) as determined by echocardiography in diastole, but amlodipine treatment did not. LVMI, PWTd and IVSTd were decreased more significantly by the treatment with candesartan than by that with amlodipine (P=0.020, 0.031 and 0.043). The present study thus revealed that candesartan effectively induced regression of LVH in type 2 diabetic patients with hypertension due to effects beyond reduction in BP. PMID- 16720058 TI - Vinorelbine and cisplatin combination in pretreated patients with advanced non small cell lung cancer pretreated with a taxane-based regimen: a multicenter phase II study. AB - PURPOSE: To assess the efficacy and tolerance of the vinorelbine/cisplatin combination in non-small cell lung cancer patients pre-treated with a taxane based regimen. PATIENTS AND METHODS: Among the 32 enrolled patients, 28 (87.5%) had a PS (WHO) of 0-1 and 13 (40.6%) have previously received both platinum compounds and taxanes. Vinorelbine (25 mg/m2 on days 1 and 8) was given by a rapid i.v. infusion and cisplatin (80 mg/m2 on day 8) after appropriate hydration. The treatment was repeated every 3 weeks. RESULTS: A partial response was achieved in six patients (ORR=18.8%; 95% confidence interval: 5.23-32.27); 13 (44.8%) and 10 (34.5%) patients had stable and progressive disease, respectively (intention-to-treat analysis). Four partial responses were observed in patients who were previously treated with taxanes/platinum-containing regimens. The median time to tumor progression was 4.7 months (range, 1.3-15.4). After a median follow up period of 6.3 months (range, 1.3-15.4) the median overall survival was 7.6 months and the 1-year survival rate 17.7%. Grade 3 and 4 granulocytopenia was observed in 11 (34.4%) patients and grade 4 thrombocytopenia in one (3.1%). Eleven (34.4%) patients presented grade 2 and 3 anemia. Febrile neutropenia occurred in one (3.1%) patient. Grade 3 and 4 nausea/vomiting was reported in one (9.3%) patient each and grade 2 fatigue in four (12.5%). CONCLUSIONS: The combination of vinorelbine and cisplatin is an active and well tolerated salvage regimen in NSCLC patients pre-treated with taxane-based chemotherapy. PMID- 16720059 TI - Hypoxia increases endothelin-1 mRNA expression but not immunoreactive endothelin in the medium of T98G glioblastoma cells under cytokine treatment. AB - Endothelin-1 (ET-1) levels in the culture medium were considered to reflect the transcription of the ET-1 gene and the subsequent secretion of ET-1 from cultured cells. It has not been clarified how different ET-1 mRNA expression levels and immunoreactive (IR)-ET levels in the culture medium are in the cell culture system. We studied ET-1 mRNA expression levels and IR-ET levels in the medium of T98G glioblastoma cells treated with cytokines. T98G glioblastoma cells were cultured with cytokines (interferon-gamma 100 U/ml, tumor necrosis factor-alpha 20 ng/ml and interleukin-1beta 10 ng/ml) under normoxia or hypoxia (1% O(2)). Northern blot analysis showed that ET-1 mRNA expression levels were increased by tumor necrosis factor-alpha alone or a combination of tumor necrosis factor-alpha and interleukin-1beta, or three cytokines, and the increase was further enhanced under hypoxia. Particularly, relative expression levels of ET-1 mRNA were significantly higher under hypoxia than in normoxia in the treatment with a combination of three cytokines. IR-ET levels in the medium were increased by treatment with tumor necrosis factor-alpha, interleukin-1beta or a combination of tumor necrosis factor-alpha and interleukin-1beta, or three cytokines. In contrast to the mRNA expression levels, IR-ET levels in the medium of T98G cells treated with a combination of three cytokines were rather decreased under hypoxia compared with those in normoxia. These findings indicate that hypoxia induces ET 1 mRNA expression in the treatment of three cytokines, but IR-ET levels in the medium do not reflect this induction in T98G glioblastoma cells. PMID- 16720060 TI - Fragment 31-35 of beta-amyloid peptide induces neurodegeneration in rat cerebellar granule cells via bax gene expression and caspase-3 activation. A crucial role for the redox state of methionine-35 residue. AB - The amyloid beta-peptide (AbetaP) is the major protein component of brain senile plaques in Alzheimer's disease. The redox state of methionine-35 residue plays a critical role in peptide neurotoxic actions. We used the fragment 31-35 of AbetaP [AbetaP(31-35)], containing a single methionine-35 residue (Met-35), to investigate the relationship between the oxidative state of Met-35 and neurotoxic and pro-apoptotic actions induced by the peptide; in rat cerebellar granule cells (CGC), we compared the effects of AbetaP(31-35), in which the Met-35 is present in the reduced state, with those of a modified peptide with oxidized Met-35 [AbetaP(31-35)Met-35(OX)](,) as well as an AbetaP-derivative with Met-35 substituted by norleucine [AbetaP(31-35)Nle-35]. AbetaP(31-35) induced a time dependent decrease in cell viability. AbetaP(31-35)Met-35(OX) was significantly less potent, but still induced a significant decrease in cell viability compared to control. No toxic effects were observed after treatment with AbetaP(31-35)Nle 35. AbetaP(31-35) induced a 2-fold increase in bax mRNA levels after 4h, whereas AbetaP(31-35)Met-35(OX) raised bax mRNA levels by 41% and AbetaP(31-35)Nle-35 had no effect. Finally, AbetaP(31-35) caused a 43% increase in caspase-3 activity after 24h; AbetaP(31-35)Met-35(OX) caused only a 18% increase, and AbetaP(31 35)Nle-35 had no effect. These findings suggest that AbetaP(31-35)-induced neurodegeneration in CGC is mediated by a selective early increase in bax mRNA levels followed by delayed caspase-3 activation; the redox state of the single Met-35 residue is crucial in the occurrence and extent of the above phenomena. PMID- 16720061 TI - Vaccinia scars associated with better survival for adults. An observational study from Guinea-Bissau. AB - BACKGROUND: Live vaccines including BCG and measles may have non-targeted beneficial effects on childhood survival in areas with high mortality. The authors therefore undertook a survey of vaccinia scars to evaluate subsequent mortality. SUBJECTS: Based on a population census, a cohort of 1893 adults in urban Guinea-Bissau was examined in 1998 and followed until 2002. MAIN OUTCOME MEASURE: All cause mortality, excluding accidents. RESULTS: The median age of vaccinia vaccinations had been 16-18 years. Adults with a vaccinia scar had a mortality ratio (MR) of 0.60 (0.41-0.87) compared to those without any scar. The effect was stronger for women. Mortality decreased with each additional vaccinia scar (MR=0.73 (0.56-0.95)). Among 502 individuals with information on HIV infection, the age-adjusted HIV-2 prevalence was 2.45 (1.06-5.65) for those with a vaccinia scar. Control for district, ethnic group, schooling, place of birth, quality of housing and HIV status had little effect on the estimate. Since vaccinia and BCG scars could have been confused, mortality for adults with vaccinia and/or BCG scar was compared to those without, the MR being 0.61 (0.41 0.89). CONCLUSION: Known cultural or socio-economic factors possibly associated with access to vaccination had no influence on the mortality ratio for having a vaccinia scar. Hence, vaccinia vaccination may have a prolonged beneficial effect on adult survival. PMID- 16720062 TI - Effectiveness of influenza vaccination in preventing influenza-like illness among community-dwelling elderly: population-based cohort study in Japan. AB - A population-based cohort study was conducted during the 2003-2004 season to examine the effectiveness of influenza vaccine among community-dwelling elderly. The subjects consisted of 4787 elderly, ranging from 65 to 79 years. We either interviewed the elderly directly or their families regarding acute febrile illness, hospital visits, hospitalization and death by telephone every month. The vaccination status and physician-diagnosed clinical influenza (hereinafter referred as clinical influenza) were determined based on data obtained from the city office and hospitals, respectively. Influenza-like illness (ILI) was defined as an acute febrile illness (> or = 38.5 degrees C) during the epidemic period. After adjusting for confounders, vaccination decreased ILI significantly (odds ratio [OR], 0.38; 95% confidence interval [CI], 0.17-0.85), but not clinical influenza (OR, 0.76; 95% CI, 0.28-2.06). The results were inconclusive for preventing hospitalization for influenza or pneumonia (OR, 0.37; 95% CI, 0.09 1.47) and death (OR, 3.68; 95% CI, 0.75-18.12), due to the inadequate sample size. In conclusion, the influenza vaccination was thus found to be associated with a decreased ILI during the epidemic period in community-dwelling elderly. PMID- 16720063 TI - Overexpression of heat shock protein 27 is associated with good prognosis in the patient with oral squamous cell carcinoma. AB - We investigated the expression and biological significance of heat shock protein (HSP)-27 in patients with oral squamous cell carcinoma (SCC). The expression of HSP-27 was quantified immunohistochemically in specimens from 37 patients with oral SCC. Findings were correlated with lymph node metastases, effect of chemotherapy, and survival. The presence of HSP-27 was identified in 31 of the 37 specimens (84%). Expression was low in 4 patients (11%), intermediate in 13 (35%), and high in 14 (38%). There were significant differences in the therapeutic effect of chemotherapy (Oboshi-Shimosato's grade) and prognosis in relation to expression of HSP-27. We found no correlation between the extent of expression of HSP-27 and stage or differentiation of the tumour. PMID- 16720064 TI - Total mercury and mercury species in birds and fish in an aquatic ecosystem in the Czech Republic. AB - Total mercury and mercury species (methylmercury-MeHg, inorganic mercury--Hg(2+)) were determined in the aquatic ecosystem Zahlinice (Czech Republic). Four tissues (muscle, intestines, liver and kidney) of three bird species--cormorant, great crested grebe and Eurasian buzzard, muscle tissues of common carp, grass carp, northern pike, goldfish, common tench, perch and rudd, aquatic plants (reed mace and common reed), sediments and water were analysed. Relative contents of MeHg (of total Hg) were in the range from 71% to 94% and from 15% up to 62% in the muscle and intestines and in liver, respectively, for all birds. Statistically significant differences were found between contents of MeHg in liver tissues of young and adult cormorant populations (F(4.60)=56.71, P<10(-5)). Relative contents of MeHg in muscle tissues of fishes were in the range from 65.1% to 87.9% of total Hg. PMID- 16720065 TI - Epiphytic lichens as biomonitors of atmospheric pollution in Slovenian forests. AB - Two country-wide surveys using epiphytic lichens as biomonitors of atmospheric pollution carried out during 2000 and 2001 in Slovenia were compared with surveys in 1991 and 1992. In the first survey, epiphytic lichen cover was studied in more than 500 plots of the 4 x 4 km national grid carried out within the framework of forest decline inventories. In the second survey, the epiphytic lichen Hypogymnia physodes (L.) Nyl., was collected on a 16 x 16 km bioindication grid and analysed for S, N, As, Br, Ce, Cd, Cr, K, La, Mo, Rb, Sb, Th, U and Zn contents. Only 'forested area' sampling points were included in the present study. Lichen cover was low, with about 70% of plots with less than 10% foliose lichen cover. No relationship was found between Hypogymnia trace element, N and S concentrations and foliose epiphytic lichen cover. PMID- 16720066 TI - Effects of copper on early developmental stages of Lessonia nigrescens Bory (Phaeophyceae). AB - Copper effects on the early developmental gametophytic and sporophytic stages of the kelp Lessonia nigrescens were tested in gradients of increasing concentrations of ASV-labile copper. The results demonstrated a high sensitivity to copper of all life-history stages of the alga, where even the lowest tested concentration affected spore release as well as their subsequent settlement. More significant, concentrations higher than 7.87 microg L(-1) totally interrupted the development of the spores after they settle. This effect led to a failure in the formation of male and female gametophytes and, as a consequence, to a complete disruption of the normal life cycle of the kelp. Thus, we suggest that the absence of L. nigrescens from copper-enriched environments results from the high sensitivity of its early life cycle stages, which limits growth and maturation of the gametophytic microscopic phase and, as a consequence, prevents development of the macroscopic sporophytic phase. PMID- 16720067 TI - Assessment of a sewage sludge treatment on cadmium, copper and zinc bioavailability in barley, ryegrass and earthworms. AB - The toxicity and bioavailability of metals were assessed to verify the efficiency of a new chemical leaching process (METIX-AC) to minimize the risk of metals found in municipal sewage sludge. For this purpose, sludge samples were spiked with cadmium, copper and/or zinc before being treated using METIX-AC. The sludge decontamination resulted in a removal of spiked metals (79-89%), in a decrease of the more labile fractions, and in a corresponding increase of the residual fraction. The toxicity observed after exposure of two plant species, barley (Hordeum vulgare) and ryegrass (Lolium perenne), and a terrestrial invertebrate, Eisenia andrei, to sludge-soil mixtures, disappeared after treatment, although the adverse effects were minor before treatment. The sludge treatment also significantly decreased the bioaccumulation of cadmium, copper, and zinc in the exposed species. For cadmium, maximum tissue concentrations of 0.45+/-0.08 mg/kg in barley, 0.79+/-0.27 mg/kg in ryegrass, and 21.82+/-1.85 mg/kg in earthworm exposed to sludge before treatment decreased after treatment to values similar to those observed with negative controls. PMID- 16720068 TI - Possible neuroleptic malignant syndrome related to concomitant treatment with paroxetine and alprazolam. AB - A 74-year-old man with depressive symptoms was admitted to a psychiatric hospital due to insomnia, loss of appetite, exhaustion, and agitation. Medical treatment was initiated at a daily dose of 20 mg paroxetine and 1.2 mg alprazolam. On the 10th day of paroxetine and alprazolam treatment, the patient exhibited marked psychomotor retardation, disorientation, and severe muscle rigidity with tremors. The patient had a fever (38.2 degrees C), fluctuating blood pressure (between 165/90 and 130/70 mg mm Hg), and severe extrapyramidal symptoms. Laboratory tests showed an elevation of creatine phosphokinase (2218 IU/L), aspartate aminotransferase (134 IU/L), alanine aminotransferase (78 IU/L), and BUN (27.9 mg/ml) levels. The patient received bromocriptine and diazepam to treat his symptoms. 7 days later, the fever disappeared and the patient's serum CPK levels were normalized (175 IU/L). This patient presented with symptoms of neuroleptic malignant syndrome (NMS), thus demonstrating that NMS-like symptoms can occur after combined paroxetine and alprazolam treatment. The adverse drug reaction score obtained by the Naranjo algorithm was 6 in our case, indicating a probable relationship between the patient's NMS-like adverse symptoms and the combined treatment used in this case. The involvement of physiologic and environmental aspects specific to this patient was suspected. Several risk factors for NMS should be noted in elderly depressive patients whose symptoms often include dehydration, agitation, malnutrition, and exhaustion. Careful therapeutic intervention is necessary in cases involving elderly patients who suffer from depression. PMID- 16720069 TI - Analysis of inhibition, reactivation and aging kinetics of highly toxic organophosphorus compounds with human and pig acetylcholinesterase. AB - Organophosphorus compounds (OP) are in wide spread use as pesticides and highly toxic OP may be used as chemical warfare agents (nerve agents). OP inhibit acetylcholinesterase (AChE), therefore, standard treatment includes AChE reactivators (oximes) in combination with antimuscarinic agents. In the last decades, the efficacy of oximes has been investigated in various animal models, mostly in rodents. However, extrapolating animal data to humans is problematical because of marked differences between rodents and humans concerning the toxicokinetics of nerve agents, the pharmacokinetics of antidotes and the AChE enzyme kinetics. In order to improve the understanding of species differences and to enable a more reliable extrapolation of animal data to humans a study was initiated to investigate the effect of highly toxic nerve agents, i.e. VX, Russian VX (VR) and Chinese VX (CVX), with human and pig erythrocyte AChE. Hereby, the rate constants for the inhibition of AChE by these OP (ki) and for the spontaneous dealkylation (ka) and reactivation (ks) of OP-inhibited AChE as well as for the oxime-induced reactivation of OP-inhibited AChE by the oximes obidoxime, 2-PAM, HI 6, HLo 7 and MMB-4 were determined. Compared to human AChE pig AChE showed a lower sensitivity towards the investigated OP. Furthermore, a slower spontaneous dealkylation and reactivation of pig AChE was recorded. The potency of the investigated oximes was remarkably lower with OP-inhibited pig AChE. These data may contribute to a better understanding of species differences and may provide a kinetic basis for extrapolation of data from pig experiments to humans. PMID- 16720070 TI - Cardio protective effect of glucose-insulin infusion on acute digoxin toxicity in rat. AB - We recently observed a case of digoxin and insulin self-poisoning without cardiac repercussion. We raised the hypothesis that insulin may have a cardio-protective effect in case of digoxin toxicity. We have therefore evaluated the effect of glucose-insulin infusion on mortality and ECG abnormalities during acute digoxin toxicity in rats. Before and after a hyperinsulinemia-euglycemia clamp, rats in glucose-insulin-digoxin (GID) group (n=10) received an intravenous infusion of 12ml/h or 2,5ml/h digoxin (0.25mg/ml) respectively until death occured. Animals receiving digoxin or saline solution intravenously served as control (n=10). ECG recording was performed in all animals over the entire period. Serum insulin and digoxin concentrations were measured by ELISA method after digoxin administration. When digoxin was administered after the clamp, all animals in GID group were alive, whereas 80% of animals in the digoxin group were dead (p<0.001) after 30min. The administration of Digoxin provoked rapid death of rats in the digoxin group in 15+/-12min whereas in GID group the survival period was significantly increased to 38+/-3min (p<0.001). Twenty minutes after digoxin administration, P waves disappeared for 78% of animals in digoxin group while they were present in all rats of GID group (p<0.001). Animal death occurred after a digoxin infusion volume of 7.7+/-0.6ml and 3.0+/-2.4ml in GID and digoxin group respectively (p<0.001). Five minutes after digoxin administration, potassium plasmatic level increased significantly in digoxin group as compared to GID group: 7.1+/-2mmol/l versus 4.4+/-0.4mmol/l (p<0.001). When digoxin was infused before the clamp, 40% of animals in GID group were alive after 180min and the other 60% died after 137+/-40min whereas death of rats in the digoxin group occurred within 80+/-10min (p<0.001). The death of animals was preceded by the P waves disappearing. Thirty minutes after digoxin administration, the potassium plasmatic level increased significantly in the digoxin group as compared to the GID group: 6.9+/-0.5mmol/l versus 4.9+/-0.3mmol/l (p<0.001). At the time of death, both volume of digoxin infusion and serum digoxin concentration were increased in GID group as compared to digoxin group: 5.7+/-1.6ml versus 3.3+/ 0.4ml (p<0.001) and 10.7+/-8.3mg/l versus 8.5+/-4.6mg/l. CONCLUSION: Glucose insulin infusion delayed the abnormalities in cardiac conduction and improved rat survival after acute digoxin toxicity. These results suggest a cardioprotective effect of insulin in case of acute digoxin toxicity. PMID- 16720071 TI - Effects of social interaction on monocular/unihemispheric sleep in male and female domestic chicks. AB - The monocular/unihemispheric sleep was studied in male and female domestic chicks reared socially. In first week post-hatching, chicks were reared in couples whilst during the second week, for half of the chicks the cospecific was maintained (Conspec-chicks) and for the other half the conspecific was removed (No-Conspec-chicks). During the first week, female chicks showed a bias for more left-eye closure/right unihemipheric sleep, whilst male chicks did not show any eye-closure bias. In the second week, both female and male Conspec-chicks showed a bias for right eye-closure/left unihemispheric sleep. Female No-Conspec-chicks did not show any eye-closure bias whilst male No-Conspec-chicks showed a bias for more left eye-closure/right unihemispheric sleep. Based on the role of the avian brain lateralization, a bias for more right or left eye-closure could be associated with a prevalent activation of left or right hemisphere during wakefulness. Eye-opening during sleep might be connected with the hemisphere that was not or was less activated during wakefulness or with lateralization of the environment monitoring against predation. PMID- 16720072 TI - Serum lipid oxidizibility in term premature rupture of the membranes. AB - OBJECTIVE: In our previous studies we have shown that the process of term labor is associated with oxidative stress, as indicated by increased susceptibility of maternal serum lipids to copper induced peroxidation. In order to continue evaluating the role of oxidative stress in the labor process, we next tested whether term premature rupture of the membranes (PROM) is also associated with increased susceptibility of maternal serum lipids to copper induced peroxidation. DESIGN: A controlled prospective study. SETTING: Tertiary care centre. POPULATION: 31 healthy women with term PROM and 19 healthy pregnant women with intact membranes. The women were matched for maternal and gestational age. METHODS: Venous blood was drawn from the women (up to 6h after rupture of the membranes and prior to labor in the PROM group), and the kinetics of copper induced oxidation of serum lipids ex vivo were monitored spectroscopically at 37 degrees C by continuous recording of absorbance at 245 nm. RESULTS: The lag phase, reflecting resistance of serum lipids to oxidation, was similar in the PROM group when compared to the control group (43.7+/-3.2 versus 41.9+/-1.6 min, P=0.61). However, the maximal rate of oxidation (V(max)) and the maximal accumulation of absorbing products (OD(max)) were shorter in the PROM group when compared to the control group (5.14+/-0.26 versus 6.29+/-0.4010(-3) OD(245) nm/min, P=0.016; 0.61+/-0.03 versus 0.71+/-0.04 OD(245) nm, P=0.07). CONCLUSION: As opposed to term labor, term PROM is not associated with increased maternal systemic oxidative stress when compared to normal pregnant women. The role for oxidative stress in preterm PROM warrants further studies. PMID- 16720073 TI - Comparisons of follicular levels of sex steroids, gonadotropins and insulin like growth factor-1 (IGF-1) and epidermal growth factor (EGF) in poor responder and normoresponder patients undergoing ovarian stimulation with GnRH antagonist. AB - OBJECTIVE: To determine the serum and intrafollicular concentrations of sex steroids, epidermal growth factor (EGF) and insulin like growth factor-1 (IGF-1) in women demonstrating poor response to ovarian stimulation with gonadotropins and GnRH antagonists, and to compare the results with age-matched women displaying normal ovarian response. STUDY DESIGN: This is a prospective cross sectional study conducted in a private IVF center. Forty-eight age-matched women producing 5 or fewer oocytes (poor responders) or 10 or more oocytes (normoresponders) at the end of controlled ovarian stimulation for assisted conception participated in the experiment. Gonadotropins and GnRH antagonists were used for ovarian stimulation, while ICSI was employed for assisted fertilization. Serum and follicular concentrations of FSH, LH and sex steroids (estradiol, progesterone and testosterone), and follicular concentrations of EGF and IGF-1 were assayed in both groups. RESULTS: Serum and follicular levels of E(2) and progesterone were significantly lower in the poor responder group compared to the normoresponder group. Follicular level of testosterone was also lower in poor responders, but not to a level of statistical significance. The serum FSH level was higher in the poor responder group, but follicular levels of gonadotropins did not differ between the two groups. The follicular level of IGF 1 was significantly lower in poor responders. In contrast, the EGF concentration did not differ between the two groups. CONCLUSIONS: Decreased levels of sex steroids in poor responder patients undergoing COH with GnRH antagonist, suggests that reduced IGF-1 expression acts as a modulator of impaired ovarian steroidogenesis. PMID- 16720074 TI - Predictive value of a single early fetal weight estimate in normal pregnancies. AB - OBJECTIVES: To evaluate the accuracy of ultrasound-based fetal weight estimates made at 28-34 weeks of gestation in predicting small- and large-for-gestational age infants (SGA, LGA) at term. METHODS: Two-hundred and fifty-nine patients with a healthy, singleton pregnancy in whom fetal biometry measurements were routinely performed between 28 and 34 weeks' gestation, were recruited at term delivery. The sonographic estimated fetal weight (EFW) and the birth weight were converted to percentiles on the basis of locally developed growth charts and compared. Multivariate linear stepwise regression analysis was used to predict the birth weight and birth weight percentile. The resulting equation (projectile formula) was used to determine the calculated birth weight, and that value was compared with the actual birth weight. The Bland and Altman plot and Passing and Bablok regression were used to compare between the calculated birth weight and the actual birth weight. RESULTS: Mean gestational age at ultrasound examination was 32+/-1.6 weeks (28-34), and mean age at delivery was 39+/-1.7 weeks (37-42). The multivariate correlation between the calculated birth weight and the birth weight (R2 = 0.524) was higher than the correlation between the sonographic EFW and the birth weight (R2 = 0.083). Both the sonographic EFW and the calculated birth weight are characterized by low positive predictive values in predicting SGA or LGA infants. The calculated birth weight was more accurate in excluding SGA and LGA infants (negative predictive values of 99.5% and 100%, respectively). On method comparison tests, the calculated birth weight was not significantly different than the actual birth weight. CONCLUSIONS: Fetal weight estimation at the early third trimester poorly predicts the birth weight centile at term. It remains uncertain, however, if it would be useful to use the calculated birth weight in pregnancies with clinically suspected SGA or LGA fetuses. PMID- 16720075 TI - Hemodynamic changes of renal main arteries in pregnancy-induced hypertension. AB - OBJECTIVE: To detect Doppler ultrasonographic velocimetry alterations of renal main arteries in pregnancy-induced hypertension (PIH). STUDY DESIGN: The peak systolic and end-diastolic velocities (Vs, Vd), resistivity and pulsatility indices (RI, PI), systolic/diastolic ratio (S/D ratio), acceleration time (AT) and systolic acceleration (SA) of renal main arteries were obtained in 17 PIH women and 15 age- and gestation week-matched normal pregnant (NP) women. The data between the two groups were compared with unpaired student's t-test. RESULTS: The AT was much longer and SA much lower in PIH than those in NP, while no significant differences were found in Vs, Vd, RI, PI and S/D ratio. CONCLUSIONS: AT and SA could be better used than Vs, Vd, RI, PI or S/D for detection of hemodynamic changes of renal arteries in patients with PIH. It may also imply that larger arteries proximal to the renal main artery be significantly affected. PMID- 16720077 TI - A new artificial urinary sphincter with conditional occlusion for stress urinary incontinence: preliminary clinical results. AB - OBJECTIVES: To perform a preliminary clinical investigation to determine the safety and efficacy of a novel artificial urinary sphincter (AUS) with conditional occlusion for the treatment of stress urinary incontinence. METHODS: Male patients with urodynamically proven stress urinary incontinence after a prostatectomy were implanted with the novel AUS. They were followed up over a period of 12 months and the device tested for efficacy by using objective measurements of urinary leakage and continence. We derived a new measure for continence called the Continence Index. RESULTS: We have demonstrated that the patients receiving the new AUS showed a reduction of greater than 10-fold in mean daily leakage volume from 770.6 ml to 55.1 ml. There was an overall improvement in the Continence Index from 54% to 97%. CONCLUSIONS: The new AUS with conditional occlusion provides good continence rates and enables adjustment of regulating pressure in situ. PMID- 16720078 TI - Female sexual maturation and reproduction after prepubertal exposure to estrogens and endocrine disrupting chemicals: a review of rodent and human data. AB - Natural hormones and some synthetic chemicals spread into our surrounding environment share the capacity to interact with hormone action and metabolism. Exposure to such compounds can cause a variety of developmental and reproductive detrimental abnormalities in wildlife species and, potentially, in human. Many experimental and epidemiological data have reported that exposure of the developing fetus or neonate to environmentally relevant concentrations of some among these endocrine disrupters induces morphological, biochemical and/or physiological disorders in brain and reproductive organs, by interfering with the hormone actions. The impact of such exposures on the hypothalamic-pituitary gonadal axis and subsequent sexual maturation is the subject of the present review. We will highlight epidemiological human studies and the effects of early exposure during gestational, perinatal or postnatal life in female rodents. PMID- 16720079 TI - Levels of detection of Cryptosporidium oocysts in mussels (Mytilus galloprovincialis) by IFA and PCR methods. AB - Cryptosporidium spp. are monoxenous protozoan parasites that cause gastrointestinal diseases in humans and animals. Shellfish harvesting areas can become contaminated by the infectious stage of the parasite and humans are therefore at risk of infection either by consumption of shellfish, or by taking part in recreational activities in these areas. In the present study we determined the levels of detection, by IFA and PCR techniques, of Cryptosporidium oocysts in mussels experimentally contaminated with a theoretical number of oocysts. There was a significant correlation between the results obtained by both techniques (P<0.05). IFA and PCR were also applied to a total of 222 samples of mussels (Mytilus galloprovincialis) destined for human consumption. In the naturally contaminated samples, we detected a 31.1% of contamination and only Cryptosporidium parvum (previously denominated C. parvum genotype II) was identified. PMID- 16720080 TI - Epidemiology of skin cancer arisen from the burn scars in Nigerian Ibos. AB - During the period 20 February 1970-19 February 2000, burns resulting in squamous cell carcinoma of the skin were documented by using a histopathology data pool of surgical specimens kept by the author as regards his Ibos ethnic group in Nigeria, West Africa. There were 21 cases. The males outnumbered the females in the ratio of 3:1. The youngest patient was aged 8 years and the oldest 75 years (mean age 39 years). Most of the antecedent injuries occurred during childhood. The two etiologic agents of albinism and burns were combined in one patient while another rarity was the presentation of the cancer within keloids. In conclusion, in dark skinned races, research should be directed on the comparative role of burns in predisposing to squamous cell carcinoma in individuals whose skin is compromised by either albinism or keloids. PMID- 16720081 TI - Pattern of use and subjective effects of Salvia divinorum among recreational users. AB - BACKGROUND: Salvia divinorum is a member of the Lamiaceae family and contains the psychotropic diterpene and kappa-opioid receptor agonist salvinorin-A. Originally a shamanic inebriant used by the Mexican Mazatec Indians, the plant and its preparations are becoming increasingly popular among non-traditional users. METHODS: Demographic data and information on pattern of use and subjective effects were obtained by means of self-report questionnaires from a sample of 32 recreational users of salvia and other psychedelics. RESULTS: Involvement with salvia appeared to be a recent phenomenon. Smoking the extract was the preferred form of administration. Subjective effects were described as intense but short lived, appearing in less than 1 min and lasting 15 min or less. They included psychedelic-like changes in visual perception, mood and somatic sensations, and importantly, a highly modified perception of external reality and the self, leading to a decreased ability to interact with oneself or with one's surroundings. CONCLUSIONS: Although some aspects of the subjective effects reported were similar to high doses of classical psychedelics with serotonin-2A receptor agonist activity, the intense derealization and impairment reported appear to be a characteristic of salvia. The observed simultaneous high scores on the LSD and PCAG subscales of the Addiction Research Center Inventory (ARCI) have been previously reported for other kappa-opioid agonists, and support kappa receptor activation as the probable pharmacologic mechanism underlying the modified state of awareness induced by salvia. PMID- 16720082 TI - Smoking status in the initial weeks of quitting as a predictor of smoking cessation outcomes in pregnant women. AB - OBJECTIVE: Any smoking during the initial 2 weeks of attempting to quit predicts poor short- and longer-term outcomes in the general population of cigarette smokers. The present study examined whether that rule applies to pregnant women. METHODS: Data were obtained from 129 women participating in clinical trials on smoking-cessation examining the efficacy of voucher-based incentives delivered contingent on biochemically-verified abstinence or a control condition wherein incentives were given independent of smoking status. Smoking status was assessed in weeks 1 and 2 of the cessation effort and again at an end-of-pregnancy assessment scheduled at weeks 28-32 gestation using self-report and biochemical verification. RESULTS: Smoking in weeks 1 or 2 predicted smoking at the end-of pregnancy assessment independent of treatment condition. There was a 79% and 92% chance that those who smoked in weeks 1 or 2 would be classified as smokers at end-of-pregnancy in the contingent-incentive and control conditions, respectively. CONCLUSIONS: Clinicians assisting pregnant women trying to quit smoking may want to monitor progress in the initial weeks of the cessation effort and consider enhancing/changing the intervention when smoking is detected. PMID- 16720083 TI - Interactions of buprenorphine and dipotassium clorazepate on anxiety and memory functions in the mouse. AB - Buprenorphine, a partial mu-receptor agonist widely substituted for heroin in the treatment of addiction, is often misused in combination with benzodiazepines. Improved hedonic properties may result, but only at the cost of increased buprenorphine toxicity. In order to elucidate the appeal of the benzodiazepine buprenorphine combination, the present study looked at its neuropsycho pharmacological effects on various emotional and cognitive parameters in the mouse. On the basis of previous dose-response studies, the regimen used was buprenorphine 0.3mg/kg, s.c. plus dipotassium clorazepate 1, 4 and 16 mg/kg, i.p. Anxiety-like behaviour was assessed using the black and white test box, and memory processes were examined via the spontaneous alternation paradigm in the Y maze, and passive avoidance tests. Spontaneous locomotor activity was also evaluated. High doses of clorazepate impaired buprenorphine-induced hyperactivity and anxiogenic-like effects. They also increased buprenorphine-induced spontaneous alternation impairment, but did not modify its impact on long-term memory processes. These results suggest that the positive reinforcement experienced with the buprenorphine-benzodiazepine combination may be attributable, at least in part, to an increase in buprenorphine's sedative effect associated with a decrease in anxiogenicity. PMID- 16720084 TI - Influence of supplemental dietary sprouted wheat on reproduction in artificially inseminated doe rabbits. AB - The sprouted wheat contains great amounts of 6-methoxybenzoxazolinone (6-MBOA) a phenol compound that stimulates reproduction in certain small wild herbivorous mammals. The objective of the present study was to evaluate the effect of dietary sprouted wheat as supplement on reproduction in artificially inseminated doe rabbits. Two-month-old New Zealand White doe rabbits (n = 48) were randomly allocated to one of two treatments: (1) pelleted diet plus sprouted wheat for 6 consecutive days prior to service, (2) pelleted diet only (control). First insemination occurred when does reach 3200g body weight and bred 6 days after parturition across 6 months period during summer and autumn. Records from 41 does completing the experiment accounted for 192 inseminations and 142 kindlings equivalent to 4.6 +/- 0.15 and 3.5 +/- 0.15 litters per doe, respectively. Mean daily intake of sprouted wheat on wet and dry matter bases were 196.1 +/- 7.5 and 75.4 +/- 2.9g, respectively. The number of young born produced per doe during the trail was significantly (P = 0.02) greater in does fed sprouted wheat (28.1 +/- 1.2 versus 23.6 +/ -1.3 control). Does fed sprouted wheat had 0.65 +/- 0.06 receptivity rate at AI over 28 per cent greater (P = 0.001) than does in the control treatment. Sexual receptivity was not influenced by physiological status of does and season (P > 0.05). Diet and season had no effect (P>0.05) on kindling rate. However, physiological status had an effect (P = 0.002) on this variable. Kindling rates for nulliparous, lactating and non-lactating does were 0.95 +/- 0.08, 0.63 +/- 0.04 and 0.78 +/ -0.11, respectively. Sexually receptive does had a greater (P = 0.0001) kindling rate (0.95 +/- 0.05) than non-receptive females (0.63 +/-0.05). Does fed sprouted wheat produced larger (P = 0.02) litters than those in the control group: 7.7 +/- 0.30 and 6.8 +/- 0.32, respectively. There was an effect of season (P = 0.04) on the size of the litter at birth. Largest litters were born during autumn (7.9 +/- 0.37) than during summer (6.6 +/- 0.41). Receptive does had more (P = 0.002) young in the litter (7.9 +/- 0.28) than non receptive (6.6 +/- 0.35). Feeding sprouted wheat as a source of biological 6-MBOA enhanced sexual receptivity and prolificacy in artificially inseminated doe rabbits bred in summer and autumn. PMID- 16720086 TI - Solid-state properties and crystallization behavior of PHA-739521 polymorphs. AB - PHA-739521 is an experimental compound that exhibits polymorphism. The two anhydrous crystal forms, I and II, are characterized using powder X-ray diffractometry, thermal analyses, moisture sorption gravimetry. Both Forms I and II are non-hygroscopic and are stable to compaction pressure. The melting temperature is about 152 degrees C for Form I and 168 degrees C for Form II. Forms I and II are enantiotropically related where Form I is more stable below a transition temperature of approximately 70 degrees C. Crystallization behavior of this compound from solutions and during heating is also studied. Information obtained is used to design an appropriate crystallization process to successfully manufacture desired polymorph at large scale. PMID- 16720085 TI - Simultaneous modulation of transport and metabolism of acyclovir prodrugs across rabbit cornea: An approach involving enzyme inhibitors. AB - The aim of this study is to identify the class of enzymes responsible for the hydrolysis of amino acid and dipeptide prodrugs of acyclovir (ACV) and to modulate transport and metabolism of amino acid and dipeptide prodrugs of acyclovir by enzyme inhibitors across rabbit cornea. l-Valine ester of acyclovir, valacyclovir (VACV) and l-glycine-valine ester of acyclovir, gly-val-acyclovir (GVACV) were used as model compounds. Hydrolysis studies of VACV and GVACV in corneal homogenate were conducted in presence of various enzyme inhibitors. IC(50) values were determined for the enzyme inhibitors. Transport studies were conducted with isolated rabbit corneas at 34 degrees C. Complete inhibition of VACV hydrolysis was observed in the presence of Pefabloc SC (4-(2-aminoethyl) benzenesulfonyl-fluoride) and PCMB (p-chloromercuribenzoic acid). Similar trend was also observed with GVACV in the presence of bestatin. IC(50) values of PCMB and bestatin for VACV and GVACV were found to be 3.81+/-0.94 and 0.34+/-0.08muM respectively. Eserine, tetraethyl pyrophosphate (TEPP) and diisopropyl fluorophosphate (DFP) also produced significant inhibition of VACV hydrolysis. Transport of VACV and GVACV across cornea showed decreased metabolic rate and modulation of transport in presence of PCMB and bestain respectively. The principle enzyme classes responsible for the hydrolysis of VACV and GVACV were carboxylesterases and aminopeptidases respectively. Enzyme inhibitors modulated the transport and metabolism of prodrugs simultaneously even though their affinity towards prodrugs was distinct. In conclusion, utility of enzyme inhibitors to modulate transport and metabolism of prodrugs appears to be promising strategy for enhancing drug transport across cornea. PMID- 16720087 TI - Allele frequencies of six miniSTR loci (D10S1248, D14S1434, D22S1045, D4S2364, D2S441 and D1S1677) in a Spanish population. AB - Allele frequencies and forensic parameters for six miniSTR autosomal loci (D10S1248, D14S1434, D22S1045, D4S2364, D2S441 and D1S1677) were obtained from a sample of 264 unrelated individuals from Spain. No significant deviations from Hardy-Weinberg expectations were found. Due to the small PCR products (<125 bp), the use of these non-CODIS (NC) miniSTRs can increase the probability that a degraded sample can be typed. Additionally, these systems can be used in routine paternity analyses where more markers are needed to increase the power of exclusion or in complex paternity cases (e.g. involving closely related individuals). PMID- 16720088 TI - Pneumocephalus secondary to mastoid surgery: a case report. AB - Pneumocephalus secondary to mastoid surgery is uncommon. Serious sequelae and central nervous system infection may result from inadequate management. It is diagnosed by careful history, physical examination and imaging studies. Treatments of pneumocephalus secondary to mastoid surgery are conservative treatment or surgery. The aim of surgery is to detect communication between mastoid cavity and brain, and to correct it. We present a case of pneumocephalus secondary to mastoid surgery in 22-year-old man, which was treated with surgery. PMID- 16720089 TI - Guided self-determination improves life skills with type 1 diabetes and A1C in randomized controlled trial. AB - OBJECTIVE: To report 1-year results of newly developed method, guided self determination (GSD), applied in group training (GSD-GT) for Type 1 diabetes patients with persistent poor glycaemic control. METHODS: GSD was designed on the basis of qualitative research to help patients develop life skills with diabetes using worksheets filled in at home and coached by nurses in mutual reflection. We randomized 18-49-year-old adults at a Danish university hospital to either 16 h GSD-GT in 2001 or to similar training 1 year later. INCLUSION CRITERIA: mean A1C> or =8.0% for at least 2 years, disease onset < or =40 years and insulin treatment from onset. RESULTS: Thirty GSD-GT patients and 20 controls completed the study. GSD-GT patients did better than control patients in terms of (a) increased autonomy support perceived from health professionals (p<0.01); (b) higher frequency of self-monitored blood glucoses (p<0.001); (c) increased perceived competence in managing diabetes (p<0.01); (d) fewer diabetes-related problems (p<0.05); and (e) improved glycaemic control (p<0.01). CONCLUSION: GSD was effective in improving life skills with diabetes, including A1C, over a period of 1 year. PRACTICE IMPLICATIONS: GSD is a worthy candidate for further research. We consider it adjustable to people with type 2 diabetes and other chronic conditions. PMID- 16720090 TI - A comparison of transhiatal and transthoracic resections on the prognosis in patients with squamous cell carcinoma of the esophagus. AB - AIM: The aim of this study was to investigate the long-term prognosis for squamous cell carcinoma of the esophagus treated either by the transhiatal (TH) or by the transthoracic (TT) operative approach. PATIENTS AND METHODS: Two hundred and twenty-nine patients (median age: 56 (29-84) years) with squamous cell carcinoma of the esophagus underwent esophageal resection between September 1985 and April 2004. In 70 patients, the transhiatal approach and in 159, the transthoracic approach was applied. An extended mediastinal lymph-node dissection was only carried out in the course of the transthoracic technique. RESULTS: Demographic data and tumor stages were comparable in both groups. A significantly better long-term survival was observed in patients with transthoracic approach for those who had undergone curative procedures (R0) (24 versus 13 months), as well as for those either without (pN0) (38 versus 14 months) or with lymph-node involvement (pN1), and for those with > or =16 (=median) dissected thoracic lymph nodes (25 versus 12 months) (p<0.05*). Patients with regional lymph-node involvement (pN1) were seen to have a significant prognostic advantage in cases with more than 16 (=median), rather than less than 16 mediastinal lymph nodes resected (p=0.045*). CONCLUSION: The prognosis in patients with squamous cell carcinoma of the esophagus is influenced by the number of dissected mediastinal lymph nodes. Patients with regional lymph-node involvement appear to benefit from an extended lymphadenectomy, in spite of the higher rate of complications and mortality associated with this procedure. PMID- 16720091 TI - Pregnancy outcomes following exposure to serotonin reuptake inhibitors: a meta analysis of clinical trials. AB - Serotonin reuptake inhibitors (SRIs) are extensively used in management of clinical depression. Reports vary about the risk of these drugs during pregnancy. To determine the risk of exposure to SRIs, we pooled data from multiple clinical studies that investigated obstetrical outcomes in women exposed to this group of drugs during pregnancy. Studies were identified by search of PUBMED, OVID, and SCOPUS databases and the data were derived from 1990 to 2005 (August). Types of outcome investigated were spontaneous abortion, major malformations, cardiovascular malformations, and minor malformations. The criteria for inclusion of studies in this meta-analysis were exposure of women to any therapeutic dosage of SRI (citalopram, fluoxetine, fluvoxamine, paroxetine, sertraline) during pregnancy. Our results find that SRIs do not increase the risk of major, cardiovascular and minor malformations but do increase the risk of spontaneous abortion significantly. PMID- 16720092 TI - Maintenance of inhibitory interneurons and boutons in sensorimotor cortex between middle and old age in Fischer 344 X Brown Norway rats. AB - Ultrastructurally identified inhibitory synapses in layer II of rat sensorimotor cortex decline between middle and old age [Poe, B.H., Linville, C., Brunso Bechtold, J., 2001. Age-related decline of presumptive inhibitory synapses in the sensorimotor cortex as revealed by the physical disector. J. Comp. Neurol. 439, 65-72]. The current study investigated whether a loss or shrinkage of gamma aminobutyric acid (GABA)ergic interneurons contribute to that decline. Coronal sections from middle-aged (15-17 months) and old (25-29 months) Fischer 344 X Brown Norway male rats were immunoreacted with antibodies to the GABA synthesizing enzyme glutamic acid decarboxylase (GAD); the calcium-binding protein parvalbumin (PV), or the neuronal marker NeuN. The number of GAD immunoreactive (IR), PV-IR, and NeuN-IR cells were determined stereologically using the optical disector technique and the cross-sectional areas of GAD-IR cells were measured in layers II/III, IV, V and VI of sensorimotor cortex. Neither the number of GAD-IR or NeuN-IR cells, nor the size of GAD-IR cells, declined significantly between middle and old age. A modest decline in the PV-IR subset of inhibitory interneurons was observed, predominantly due to changes in layers V and VI. Stereological analysis of layer II/III GAD-IR boutons revealed a stability of immunocytochemically identified inhibitory terminals. Taken together, these results indicate a general maintenance of overall GABAergic neurons in sensorimotor cortex between middle and old age and the loss of ultrastructurally identified inhibitory synapses may be due to the decline of a subset of GABAergic terminals. PMID- 16720093 TI - Randomized maps for assessing the reliability of patients clusters in DNA microarray data analyses. AB - OBJECTIVE: Clustering algorithms may be applied to the analysis of DNA microarray data to identify novel subgroups that may lead to new taxonomies of diseases defined at bio-molecular level. A major problem related to the identification of biologically meaningful clusters is the assessment of their reliability, since clustering algorithms may find clusters even if no structure is present. METHODOLOGY: Recently, methods based on random "perturbations" of the data, such as bootstrapping, noise injections techniques and random subspace methods have been applied to the problem of cluster validity estimation. In this framework, we propose stability measures that exploits the high dimensionality of DNA microarray data and the redundancy of information stored in microarray chips. To this end we randomly project the original gene expression data into lower dimensional subspaces, approximately preserving the distance between the examples according to the Johnson-Lindenstrauss (JL) theory. The stability of the clusters discovered in the original high dimensional space is estimated by comparing them with the clusters discovered in randomly projected lower dimensional subspaces. The proposed cluster-stability measures may be applied to validate and to quantitatively assess the reliability of the clusters obtained by a large class of clustering algorithms. RESULTS AND CONCLUSION: We tested the effectiveness of our approach with high dimensional synthetic data, whose distribution is a priori known, showing that the stability measures based on randomized maps correctly predict the number of clusters and the reliability of each individual cluster. Then we showed how to apply the proposed measures to the analysis of DNA microarray data, whose underlying distribution is unknown. We evaluated the validity of clusters discovered by hierarchical clustering algorithms in diffuse large B-cell lymphoma (DLBCL) and malignant melanoma patients, showing that the proposed reliability measures can support bio-medical researchers in the identification of stable clusters of patients and in the discovery of new subtypes of diseases characterized at bio-molecular level. PMID- 16720094 TI - Estrogen-mediated regulation of CYP7B1: a possible role for controlling DHEA levels in human tissues. AB - The current study examines regulation of CYP7B1, a DHEA 7alpha-hydroxylase, by sex hormones. Transfection with estrogen receptor alpha and treatment with 17beta estradiol in human embryonic kidney 293 cells significantly increased CYP7B1 catalytic activity and mRNA, and stimulated a human CYP7B1 reporter gene. Transfection with estrogen receptor beta showed similar but less significant effects. In the absence of receptors, 17beta-estradiol suppressed CYP7B1 activity, suggesting that estrogenic effects may be different in cells not expressing receptors. Quantitation of CYP7B1 mRNA in adult and fetal human tissues showed markedly higher CYP7B1 mRNA levels in fetal tissues compared with the corresponding adult ones, except in the liver. This indicates a tissue specific, developmental regulation of CYP7B1 and suggests an important function for this enzyme in fetal life. DHEA secreted by fetal adrenals is an essential precursor for placental estrogen formation. Since CYP7B1 diverts DHEA from the sex hormone biosynthetic pathway, estrogen receptor-mediated up-regulation of CYP7B1 should lead to less DHEA available for sex hormone synthesis and may help to maintain normal levels of estrogens and androgens in human tissues, especially during fetal development. Regulation by estrogens may also be of importance in other processes where CYP7B1 is involved, including cholesterol homeostasis, cellular proliferation, and CNS function. PMID- 16720095 TI - Compensatory strategies during normal walking in response to muscle weakness and increased hip joint stiffness. AB - Compared to young adults, older adults exhibit a slower walking speed, smaller step length, shorter swing phase time and decreased range of motion in their lower extremity joints. The underlying mechanisms causing these gait adaptations is not well understood, with various musculoskeletal parameters being put forth as contributing factors, including increased joint stiffness and decreased isometric muscle strength. The objective of this study was to identify the necessary compensatory mechanisms to overcome such musculoskeletal deficits and regain a normal walking pattern. Understanding these mechanisms has important implications for designing effective rehabilitation interventions for older adults that target specific muscle groups and properties (e.g., isometric strength versus joint stiffness) to improve gait performance. Muscle-actuated forward dynamics simulations of normal walking were analyzed to quantify compensatory mechanisms in the presence of muscle weakness in specific muscle groups and increased hip joint stiffness. Of particular importance were the compensatory mechanisms provided by the plantar flexors, which were shown to be able to compensate for many musculoskeletal deficits, including diminished muscle strength in the hip and knee flexors and extensors and increased hip joint stiffness. This importance was further highlighted when a normal walking pattern could not be achieved through compensatory action of other muscle groups when the uniarticular and biarticular plantar flexor strength was decreased as a group. Thus, rehabilitation or preventative exercise programs may consider focusing on increasing or maintaining plantar flexor strength, which appears critical to maintaining normal walking mechanics. PMID- 16720097 TI - Psychiatric disorders in patients with psychogenic non-epileptic seizures, with and without comorbid epilepsy. AB - PURPOSE: The aim of this study is to describe similarities and differences in epidemiological, psychiatric and semiologic variables between patients with psychogenic none epileptic seizures (PNES) and comorbid epilepsy (mixed PNES), and patients with PNES without comorbid epilepsy (pure PNES). RESULTS: Forty three patients with PNES diagnosed by Video-EEG were included. Twenty-four had pure PNES, and ninteen mixed PNES. Female population, age, duration of PNES, psychiatric institutionalization, psychopharmacotherapy, dissociative disorders and posttraumatic stress disorder (PTSD), were significantly higher in the pure PNES patients. Suicide attempts, antiepileptic therapy, conversive, affective and personality disorders were frequent in both groups. In the analysis of seizure semiology, the total lack of responsiveness was significantly higher in the mixed PNES group. CONCLUSIONS: Pure PNES patients showed similarities and differences in the psychiatric profile, with a greater prevalence of dissociative disorders and PTSD, factors related to psychiatric severity. PMID- 16720096 TI - Parthenolide has limited effects on nuclear factor-kappa beta increases and worsens survival in lipopolysaccharide-challenged C57BL/6J mice. AB - Parthenolide, a sesquiterpene lactone, inhibited lipopolysaccharide (LPS) stimulated nuclear factor (NF)-kappabeta and cytokine production in vitro and in rats, and improved survival in LPS challenged Swiss albino mice. We investigated whether increased survival with parthenolide was associated directly with inhibition of NF-kappabeta and cytokines in LPS challenged C57BL/6J mice. In RAW 264.7 cells, parthenolide inhibited LPS-stimulated NF-kappabeta and cytokines (interleukin [IL]-1alpha, -1beta, -2, -4, -6, and -10, interferon-gamma, tumor necrosis factor-alpha, granulocyte macrophage-colony stimulating factor, migratory inhibitory protein-1 and -2alpha, JE, and RANTES). In mice (n = 366) receiving lethal intraperitoneal (i.p.) LPS (40 mg/kg), compared to placebo, each of 5 parthenolide doses (0.25 to 4 mg/kg i.p. following LPS) reduced survival at 168h and overall worsened the hazards ratio of survival (mean +/- S.E.M.) (1.29 +/- 0.12, p = 0.04). In other mice (241), compared to saline challenge, nonlethal LPS (2.5 mg/kg) increased NF-kappabeta in lung and kidney combined and 12 of 13 plasma cytokines early (1 and 3 h) and late (6, 9 and 12 h) (p < or = 0.002 for each). Compared to nonlethal LPS, lethal LPS increased NF-kappabeta and 12 of 13 cytokines early but not significantly and late significantly (p < or = 0.05 for each). With lethal LPS, compared to placebo, parthenolide (1 mg/kg) decreased NF kappabeta and 10 of 13 cytokines early and increased NF-kappabeta and 11 of 13 cytokines late (p < or = 0.02 for early vs. late). Although parthenolide inhibits NF-kappabeta and cytokines in vitro, its effects on these mediators and survival in animal sepsis models vary. Theses differences must be understood before parthenolide or related agents are applied clinically for sepsis. PMID- 16720098 TI - Immature porcine knee cartilage lesions show good healing with or without autologous chondrocyte transplantation. AB - OBJECTIVE: The purpose of this study was to find out how deep chondral lesions heal in growing animals spontaneously and after autologous chondrocyte transplantation. METHODS: A 6mm deep chondral lesion was created in the knee joints of 57 immature pigs and repaired with autologous chondrocyte transplantation covered with periosteum or muscle fascia, with periosteum only, or left untreated. After 3 and 12 months, the repair tissue was evaluated with International Cartilage Repair Society (ICRS) macroscopic grading, modified O'Driscoll histological scoring, and staining for collagen type II and hyaluronan, and with toluidine blue and safranin-O staining for glycosaminoglycans. The repair tissue structure was also examined with quantitative polarized light microscopy and indentation analysis of the cartilage stiffness. RESULTS: The ICRS grading indicated nearly normal repair tissue in 65% (10/17) after the autologous chondrocyte transplantation and 86% (7/8) after no repair at 3 months. At 1 year, the repair tissue was nearly normal in all cases in the spontaneous repair group and in 38% (3/8) in the chondrocyte transplantation group. In most cases, the cartilage repair tissue stained intensely for glycosaminoglycans and collagen type II indicating repair tissue with true constituents of articular cartilage. There was a statistical difference in the total histological scores at 3 months (P=0.028) with the best repair in the spontaneous repair group. A marked subchondral bone reaction, staining with toluidine blue and collagen type II, was seen in 65% of all animals. CONCLUSIONS: The spontaneous repair ability of full thickness cartilage defects of immature pigs is significant and periosteum or autologous chondrocytes do not bring any additional benefits to the repair. PMID- 16720099 TI - Parkinsonian deficits in sensory integration for postural control: temporal response to changes in visual input. AB - This study investigated the effect of Parkinson's disease (PD) on the time course for postural control following the removal and reinsertion of visual information. Twelve medicated PD patients (PD) and 12 age matched control (CTRL) subjects performed two 45-s quiet standing trials, during which visual feedback was available (0-15s), deprived (15-30s), and then restored (30-45s). The 45s test trial was divided into 5s time bins to compare the time-based effect of sensory reorganization during deprivation and reintegration. Results revealed an increase in Elliptical Sway Area (ESA) following visual deprivation for both groups; this increased ESA remained significantly higher than the baseline level for the duration of the deprivation period among PD patients and returned to baseline the level among CTRL. Despite elevated ESA at the end of visual deprivation among PD patients, neither group showed a change in ESA after visual information was restored. These results indicate a PD-associated deficit with the reorganization of sensory priorities for postural control, and may implicate the basal ganglia as being critical for integration of sensory information for postural control. PMID- 16720100 TI - Commentary on the SFTG international collaborative study on the in vitro micronucleus test: to Cyt-B or not to Cyt-B? PMID- 16720101 TI - Measles transmission during commercial air travel in Brazil. PMID- 16720102 TI - The effect of performance feedback on cardiorespiratory fitness field tests. AB - We investigated the effects of performance feedback (PF) on predicting maximal oxygen uptake (VO2 max) using the 20 m Multistage Shuttle Run Test (MST) and 20 m Square Shuttle Test (SST). The agreement between these two field tests in relation to laboratory VO2 max was also examined. Forty healthy males (age: 21.5+/-2.3; BMI: 23.7+/-2.0) randomly performed four indirect VO2 max tests; that is the MST and SST, as well as a modified version of MST (MSTMD) and SST (SSTMD). During MST and SST subjects received PF with respect to both test stage and running pace. In contrast, MSTMD and SSTMD incorporated auditory feedback which solely emitted signals regulating the running pace. Participants also performed a laboratory VO2 max treadmill test (TT). ANOVA demonstrated significant mean predicted VO2 max decrements in both MSTMD (p<0.001) and SSTMD (p<0.05) compared to MST and SST, respectively. In predicting TTVO2 max, the '95% limits of agreement' analysis indicated errors equal to 3.6+/-9.6 and 1.4+/-10.3 ml kg-1 min-1 with coefficients of variation of +/-10.0% and +/-10.9%, for MST and MSTMD, respectively. The corresponding '95% limits of agreement' values for SST and SSTMD were 0.1+/-5.0 and -1.1+/-6.1 ml kg-1 min-1 with coefficients of variation of +/-5.4% and +/-6.7%, respectively. It is concluded that the application of PF leads to superior field testing performances. PMID- 16720103 TI - Calling on exercise scientists to participate in broader efforts to prevent falls in older adults. PMID- 16720104 TI - Structural study of two proteins SigE and ORF1 to predict their roles in the biochemical oxidation of sulfur anions via the global sulfur oxidation operon (sox). AB - Microbial redox reactions involving inorganic sulfur compounds in the environment are one of the major reactions of the global sulfur cycle. These reactions are mediated by phylogenetically diverse prokaryotes containing the sulfur oxidizing gene cluster (sox). The sox gene cluster of alpha-Proteobacteria comprises of at least 15 genes, which form two transcriptional units. Recently two new orfs, which code for proteins named, SigE and ORF1, were identified in Starkeya novella. Sequence analyses reveal that SigE protein has the signature sequence of ECF-type sigma factors and a helix-turn-helix (HTH) DNA binding motif whereas ORF1 is possibly an anti ECF-sigma factor, which also has the signature sequence of the dsr family of sulfate ion binding proteins. We employed homology modeling to construct the three-dimensional structures of these proteins. The model of SigE was docked on to its promoter DNA to investigate the favourable binding modes of the protein. Interactions of SigE with its anti-sigma factor ORF1 were also reported after docking these proteins. We also identified the putative sulfate ion binding residues of ORF1 by docking sulfate ion on to it. Our study provides a rational framework for understanding of the structural as well as the molecular basis of the mechanism of the regulation of sulfur oxidation reactions by SigE and ORF1 proteins via the sox operon. PMID- 16720105 TI - Therapeutic effect of propolis and paclitaxel on hepatic phase I and II enzymes and marker enzymes in dimethylbenz(a)anthracene-induced breast cancer in female rats. AB - Propolis, a natural beehive product has been known for centuries for a variety of beneficial traditional medicinal properties. The present study was conducted to ascertain the antineoplastic potential of propolis along with paclitaxel against experimental mammary carcinogenesis. Female Sprague Dawley rats at 55 days of age were treated with dimethylbenz(a)anthracene to induce breast cancer. Paclitaxel at a dose of 33 mg/kg body mass intraperitoneally and propolis 50 mg/kg body weight orally was administered to the experimental animals, immediately after the carcinogen treatment and continued until the termination of the study. At the end of the treatment activities of phase I and II xenobiotic metabolizing enzymes and liver marker enzymes were measured. A significant increase in carcinogen activating enzymes, cytochrome P(450), cytochrome b(5) and NADPH cytochrome C reductase with concomitant decrease in phase II enzymes, glutathione transferase and UDP-glucuronyl transferase were observed in animals with mammary cancer. Furthermore there was a significant decrease in alanine aminotransferase, aspartate aminotransferase with a sharp increase in alkaline phosphatase, acid phosphatase and 5' nucleotidase. Propolis treatment caused the activity of these enzymes return to almost normal control levels, indicating the protective effect of propolis against dimethyl benz(a) anthracene induced carcinogenesis. On the basis of the observed results propolis can be considered a promising chemotherapeutic agent and can be administered as an adjuvant with paclitaxel chemotherapy. PMID- 16720106 TI - Development of the human nucleus of the solitary tract: a cyto- and chemoarchitectural study. AB - The present study investigated the prenatal development of the cyto- and chemoarchitecture of the human nucleus of the solitary tract from 9 to 35 weeks, by using Nissl staining and immunoreactivity to calbindin, calretinin, tyrosine hydroxylase and GAP-43. The nucleus began to gain heterogeneity and show different subnuclei as early as 13 weeks, and approached cytoarchitectural maturation from 21 to 25 weeks. The subnuclear division pattern observed in the fetal nucleus of the solitary tract at 25 weeks was very similar to that of the adult. Neurons immunoreactive to calbindin first appeared in the medial gastrointestinal area of the nucleus at 13 weeks, particularly within a putative gelatinosus subnucleus, while calretinin immunoreactivity during fetal life suggested the possible presence of a central subnucleus. Tyrosine hydroxylase immunoreactive neurons were seen in the medial subdivisions of the nucleus of the solitary tract as early as 13 weeks, but the population continued to increase until 25 weeks. Strong GAP-43 immunoreactivity was also present in the nucleus of the solitary tract at 13 weeks, especially in the dorsolateral and commissural subnuclei, while at 21 weeks there was a significant decline of GAP-43 expression. Results from the chemoarchitectural study showed that the human nucleus of the solitary tract expressed various neurochemical substances at an early developmental age (13 weeks), even before cellular and neuropil maturation was fully attained. Expression of these factors may play an important role in establishment and integration of viscerosensory function in the nucleus. PMID- 16720107 TI - Association between human African trypanosomiasis and the IL6 gene in a Congolese population. AB - Despite the importance of behavioural and environmental risk factors, there are arguments consistent with the existence of a genetic susceptibility to human African trypanosomiasis (HAT). A candidate gene association study was conducted in the Democratic Republic of Congo using a family-based sample which included a total of 353 subjects (86 trios; one case and parents (n=258) and 23 families with more than one case and parents (n=95)). Polymorphisms located on the IL1alpha, IL4, IL6, IL8, IL10, TNFalpha and IFNgamma genes were genotyped after re-sequencing of the genes for extensive SNP search. The T allele of the IL6(4339) SNP was significantly associated with a decreased risk of developing the disease (p=0.0006) and a suggestive association was observed for the IL1alpha(5417 T) SNP and an increased risk of developing the disease. These results suggest that genetic variability of the IL6 and to a lesser extent the IL1alpha gene are involved in the development of HAT. For the TNFalpha and IL10 gene polymorphisms, association results obtained here were different from those we observed in another population living under different epidemiologic conditions. This underlines the complexity of the interactions existing between host genetic polymorphisms, parasite diversity and behavioural and environmental risk factors in HAT. PMID- 16720108 TI - Analysing the evolutionary history of HCV: puzzle of ancient phylogenetic discordance. AB - Though recombination is an important evolutionary strategy in RNA viruses, only two cases of HCV recombinant strains have been reported. Our objective was to analyze the evolutionary history of the HCV genotypes aiming to obtain evidence of significant phylogenetic discordance due to either recombination or selective forces leading to convergent/divergent evolution. The data support an evolutionary preservation of the interferon-resistance related genomic region (ISDR) for the genotypes 1 and 4. On the other hand, there was no evidence that recombination has occurred in the past with the possible exception of genotype 4. Moreover, it is evidenced that genotypes 3 and 10 split more recently than genotypes 6-9 and 11. This analysis reverberates a commonly found pattern in rapidly evolving viruses, that is the strongly disturbed evolutionary history which deforms the uniform distribution of the phylogenetic relationships across the genome, and introduces a conservative inference framework for approaching this kind of data. PMID- 16720109 TI - Electrochemical parameters of ethamsylate at multi-walled carbon nanotube modified glassy carbon electrodes. AB - In this paper, some electrochemical parameters of ethamsylate at a multi-walled carbon nanotube modified glassy carbon electrode, such as the charge number, exchange current density, standard heterogeneous rate constant and diffusion coefficient, were measured by cyclic voltammetry, chronoamperometry and chronocoulometry. The modified electrode exhibits good promotion of the electrochemical reaction of ethamsylate and increases the standard heterogeneous rate constant of ethamsylate greatly. The differential pulse voltammetry responses of ethamsylate were linearly dependent on its concentrations in a range from 2.0 x 10(-6) to 6.0 x 10(-5) mol L(-1), with a detection limit of 4.0 x 10( 7) mol L(-1). PMID- 16720110 TI - Simulation of electroporated cell by chronopotentiometry. AB - Chronopotentiometry on planar lipid bilayer (BLM) is proposed as a method for modeling the electrical phenomena in electroporated cell. Two techniques are discussed: constant-current and linear-current chronopotentiometry. It is proposed that the constant-current chronopotentiometry may provide basis for modeling the electroporated cell shortly after the removal of the electric field, when activity of cellular pumps counteracts ionic fluxes through the electropore and ionic channels. The linear-current method can be considered for modeling the cell in the later stage after electroporation, when energetical resources of the cell are gradually getting exhausted and the activity of pumps decreases. Based on this idea, it may be postulated that the electropore in the cell has fluctuating dynamics whose stochastic characteristics, similarly as biological channels, shows 1/f noise. The model implies that the fluctuations would disappear leaving the electropore with a constant resistance when efficiency of the pumps becomes very small. The results of chronopotentiometry also may suggest that opening time, conductivity and selectivity of the electropore can be controlled by the cell environment or membrane composition. PMID- 16720111 TI - Study on the aggregation and electrochemical properties of Rose Bengal in aqueous solution of cyclodextrins. AB - The interactions of Rose Bengal (RB) with alpha-cyclodextrin (alpha-CD), hydroxypropyl-beta-cyclodextrin (HP-beta-CD), hydroxypropyl-gamma-cyclodextrin (HP-gamma-CD), heptakis(2,3,6-tri-O-methyll)-beta-cyclodextrin (TM-beta-CD) were studied in aqueous solutions of 0.1 M KClO(4) and 0.1 M LiClO(4) by vis absorption, fluorescence spectroscopy as well as electrochemical measurements at 298 K. The spectrophometric results indicate that RB is included in all beta- and gamma-CDs forming complexes with a stoichiometry 1:1 whose stability is slightly higher in KClO(4) than in LiClO(4) solutions. The complex stability constants determined for salt-containing CD solutions are lower than those for water solutions. The complexation of RB with beta- and gamma-CD and the differences between the complexes obtained in the presence of the two salts were confirmed by an electrochemical study. PMID- 16720112 TI - Characterisation of an industrial affinity process used in the manufacturing of digoxin-specific polyclonal Fab fragments. AB - This paper describes the effect of several variables on the affinity process for the production of the FDA approved biotherapeutic product Digoxin Immune Fab (Ovine) (DigiFab, Protherics Inc., TN, USA). The study considers the effects of column re-use on matrix capacity and on the subsequent recovery of the antibody product, and the impact of varying column loading on matrix performance. The methodology used could be equally applied to assess the feasibility of using an affinity matrix for commercial scale purification of alternative antibody derived biotherapeutics. The capacity and specific Fab recovery were calculated through 24h equilibrium and mass balance studies. Results were assessed against data obtained through confocal scanning laser microscopy. Scale-down experiments produced specific Fab recoveries and purities that were comparable with those at production scale. The matrix capacity was found to be 45+/-15 mg of Fab/ml of matrix. Through the use of fluorescent DigiFab and confocal scanning techniques, Fab uptake onto single affinity bead was evaluated. Average intensity values calculated for each sample provided direct real-time, measure of Fab binding and matrix capacity. The results suggest that the affinity matrix had a limited reuse life as a drop in recovery is observed following the completion of a small number of process cycles (30% after three runs). The findings support that which is seen at the current manufacturing scale, where the affinity column is used for a limited number of runs. Results from this study can be used as a basis for future optimisation of this purification process. PMID- 16720113 TI - The effect of the presence of globular proteins and elongated polymers on enzyme activity. AB - We have studied the effect of a crowded (macromolecular) solution on reaction rates of the decarboxylating enzymes urease, pyruvate decarboxylase and glutamate decarboxylase. A variety of crowding agents were used including haemoglobin, lysozyme, various dextrans and polyethylene glycol. Enzyme reaction rates of all three enzymes show two different types of effect that separate the globular proteins from the polysaccharides/polymers. Increasing concentration of globular proteins caused a dramatic rise in enzyme activity up to 30% crowding concentration then the activity decreased, whereas the polymers caused a concentration dependent decrease in activity. The viscosities of the globular proteins were low compared to the polymers. The increased activity with proteins may be due to the decreased amount of free water, which leads to higher effective concentration of substrates, or to an increased oligomeric state by self association. The lower activities of all enzymes with all agents at high concentrations may be explained by a decrease in rates of diffusion. An increase in protein crowding (decrease in cell water content) may contribute to pathological conditions including cataract and Alzheimer's disease. PMID- 16720117 TI - Travel sizes bring portions to the forefront. PMID- 16720118 TI - After the media feeding frenzy: whither the Women's Health Initiative Dietary Modification Trial? PMID- 16720119 TI - Health coaching: a new opportunity for dietetics professionals. PMID- 16720120 TI - Talking about hunger in a land of plenty. PMID- 16720121 TI - Advances in basic behavioral research will make the most important contributions to effective dietary change programs at this time. PMID- 16720122 TI - Psychosocial correlates of fruit, vegetable, and dietary fat intake among adolescent boys and girls. AB - OBJECTIVE: This study examined whether hypothesized psychosocial correlates of behavior change (family/peer influence, pros, cons, self-efficacy, parent/child change strategies, and household eating rules) are associated with consumption of fruits, vegetables, and dietary fat among adolescent boys and girls. DESIGN: This cross-sectional study used questionnaires to assess psychosocial variables and multiple 24-hour recall interviews to assess dietary intake (daily servings of fruits and vegetables and percentage energy intake from dietary fat). SUBJECTS: In this study, 878 adolescents (53.6% female, 57.9% white, mean age 12.8 years, age range 11 to 15 years) completed questionnaires. STATISTICAL ANALYSES PERFORMED: Hierarchical linear regressions were conducted on the entire sample as well as on subgroups based on sex and age (young/old). RESULTS: Results indicated that child behavior change strategies, decisional balance, and household rules were related to percentage energy intake from total fat, whereas child behavior change strategies, family influence, and household rules were related to daily servings of fruit and vegetables. More psychosocial correlates were found for older than for younger adolescents. CONCLUSIONS: Both psychological and social correlates of adolescent eating behaviors were identified, and correlates differed somewhat by adolescent subgroup. Based on these findings, promising intervention strategies that include the following should be evaluated: helping adolescents alter decisional balance, teaching behavior-change strategies, and helping parents support children's dietary changes and institute supportive household rules. PMID- 16720123 TI - Relationship of ghrelin and leptin hormones with body mass index and waist circumference in a random sample of adults. AB - OBJECTIVE: To evaluate the relationship of ghrelin and leptin hormones with body mass index (BMI) and waist circumference in a population-based random sample of adult men and women subsequently categorized from normal weight to severely obese based on BMI criteria. The relationship between total ghrelin and leptin was also evaluated. DESIGN: A cross-sectional study was conducted on adults. METHODS: Two hundred thirty-three men (n=107) and women (n=126) between the ages of 23 and 75 years were randomly selected from a database of over 600,000 adults who had previously participated in a population-based study conducted by the University of Utah Cardiovascular Genetics Program. Items collected included height, weight, waist circumference, and fasting blood samples. Blood samples were later thawed, and plasma leptin and total ghrelin levels were analyzed with radioimmunoassay kits. RESULTS: Leptin levels were directly associated with BMI (r=0.72, P=0.001) and waist circumference (r=0.71, P=0.0001), whereas total ghrelin was inversely associated with BMI (r=-0.39, P=0.001) and waist circumference (r=-0.37, P=0.001). There were also statistically significant linear changes in means across the BMI categories for both hormones. After controlling for BMI and waist circumference, the highly significant correlation of leptin with ghrelin (r= 0.39) was reduced but still significant (r=-0.17, P=0.01). CONCLUSION: Leptin increases and ghrelin decreases were linear over the five BMI categories, suggesting there is no threshold of BMI where the hormone levels change abruptly. There remain other factors beyond current BMI and waist circumference that induce a correlation of these two hormones. Further understanding of the role that each of these hormones has in influencing appetite and body weight may provide insight into mechanisms involved with obesity. PMID- 16720124 TI - Normal-weight adults consume more fiber and fruit than their age- and height matched overweight/obese counterparts. AB - OBJECTIVES: To assess differences in dietary intake of overweight/obese subjects and sex-, age-, and height-matched controls and to identify dietary components associated with increased deposition of body fat. DESIGN/SUBJECTS: A convenience sample of 52 overweight/obese and 52 normal-weight adults matched for sex, age (+/-1 year), and height (+/-1 inch) were recruited from the local area. Dietary intake was assessed with the Block 60-item food frequency questionnaire, physical activity was measured by the Yale Physical Activity Survey, and percent body fat was measured via dual-energy x-ray absorptiometry. STATISTICAL ANALYSES PERFORMED: Independent t tests compared between-group consumption of dietary components. The ability of dietary components to predict percent body fat before and after controlling for age-, sex-, and physical activity-related energy expenditure and other macronutrients was assessed with multiple regression analyses. Spearman correlation coefficients examined relationships among nutrients, Food Guide Pyramid servings, and percent body fat. RESULTS: Overweight/obese subjects consumed more total fat, saturated fat, and cholesterol and less carbohydrate, complex carbohydrate, and dietary fiber than control subjects. Reported intake of dietary fiber was inversely related to percent body fat without (R(2)=0.052, P=0.02) and with (R(2)=0.045, P=0.013) control for potential confounding factors. Servings of fruit per day were negatively related to percent body fat (r=-0.40, P<0.01). CONCLUSIONS: These findings suggest that the composition of a diet, especially low dietary fiber and fruit intake, plays a role in the etiology of obesity. PMID- 16720125 TI - Associations of decisional balance, processes of change, and self-efficacy with stages of change for increased fruit and vegetable intake among low-income, African-American mothers. AB - OBJECTIVE: To determine the relationship between stage of change and decisional balance, processes of change, and self-efficacy variables of the Transtheoretical Model to increase fruit and vegetable consumption by low-income, African-American mothers, and to assess the usefulness of the model for intervention efforts. DESIGN: We used a cross-sectional questionnaire design with a food frequency questionnaire and a staging algorithm to classify women by stage of change before enrollment. We used scaled questionnaire items based on think-aloud data to measure self-efficacy, processes of change, and pros and cons to increasing fruit and vegetable intake and variety. SUBJECTS/SETTING: We recruited a convenience sample of low-income, African-American mothers (n=420) aged 18 to 45 years with children <12 years of age to complete questionnaires at community-based sites. STATISTICAL ANALYSES PERFORMED: Analysis of variance, principal component analysis, chi(2) tests, correlation, and regression analyses were conducted. RESULTS: Perceptions of benefits for health and planning meals were stronger for women in later stages compared with earlier stages. The same relationship was observed for the use of processes of change related to becoming aware of health benefits and engaging in enabling behaviors, and variety of fruits and vegetables consumed. Self-efficacy based on eating fruits and vegetables for various eating occasions and in difficult situations was greater for women in later stages compared to earlier stages. CONCLUSIONS: Interventions for women in earlier stages should include making women aware of health benefits, increasing self efficacy, and improving ability to make plans and engage in behaviors to increase intake. PMID- 16720126 TI - Comparison of two nutrition education approaches to reduce dietary fat intake and serum lipids reveals registered dietitians are effective at disseminating information regardless of the educational approach. AB - OBJECTIVE: The purpose of this study was to compare an educational approach based on the Stages of Change Model with usual care education in reducing dietary fat intake and serum lipids in individuals with hyperlipidemia. DESIGN: A 40-week randomized control study was conducted. Four education sessions were provided on an outpatient basis during a 1-month period with follow-up every 6 weeks. PARTICIPANTS: The sample consisted of 141 men and women with hyperlipidemia (mean age 50+/-11 years and a mean body mass index of 30+/-6) randomly assigned to one of two education interventions. INTERVENTION: The stages of change group was provided with tailored dietary activities based on their readiness to change to reduce dietary fat intake. Dietary information and activities for the usual care program were developed for those individuals ready to make a change in their diets. MAIN OUTCOME MEASURES: Serum lipids, anthropometric measures, readiness to change, dietary intake, and exercise data were assessed at baseline, 4, 16, 28, and 40 weeks. STATISTICAL ANALYSES: A repeated measures analysis of variance was used to compare differences between groups across time. RESULTS: Total cholesterol, low-density lipoprotein cholesterol, and body weight decreased significantly at 4 weeks (P<0.05) for both groups and were sustained over time, with no differences between the groups. CONCLUSIONS: The stages of change education approach was not more effective than the usual care education approach in decreasing dietary fat intake and serum lipids in a classroom setting. PMID- 16720127 TI - Complementary and alternative medicine education in dietetics programs: existent but not consistent. AB - This descriptive survey was undertaken to determine the inclusion of complementary and alternative medicine topics into undergraduate didactic dietetics education. The response rate was 34% (n=92) of all directors (N=273) of didactic and coordinated dietetics programs. Almost all programs (n=81; 88%) include complementary and alternative medicine instruction in some form in their curricula; the majority of content is integrated into already existing nutrition courses. The nutrition courses most often containing complementary and alternative medicine were medical nutrition therapy, advance nutrition, and community nutrition. Topics addressed were varied and included herbal supplements, functional foods, Native-American healing, and quackery in medicine. Most directors indicated that complementary and alternative medicine is an important component of dietetics education, yet many indicated that students are not being adequately prepared in this area. The mean familiarity of program directors with complementary and alternative medicine competencies for dietetics practice was 6 on a scale, with 10 being the most knowledgeable. Respondents also identified whether complementary and alternative medicine and dietary supplement competencies were being addressed at all in their curricula. Lack of time seemed to be the limiting factor to incorporation of complementary and alternative medicine topics into the curricula. Evidence from this study indicates that current curricula are providing some complementary and alternative medicine content, but a core of knowledge is lacking. The complementary and alternative medicine competencies for entry-level dietetics practice anticipated by 2006 will be useful in helping educators adequately meet the needs of future professionals in the area of complementary and alternative medicine. PMID- 16720128 TI - New and existing oils and fats used in products with reduced trans-fatty acid content. AB - The US Food and Drug Administration's final ruling on trans-fatty acid labeling issued in 2003 has caused a rapid transformation in the fat and oil industries. Novel ingredients and improved technologies are emerging to replace partially hydrogenated fats in foods. We present an overview of the structure and formation of trans fatty acids in foods, and a comprehensive review of the newly formulated products and current procedures practiced by the edible oil industry to reduce or eliminate trans fatty acids in response to the Food and Drug Administration's regulations mandating trans fat labeling of foods. PMID- 16720130 TI - Stages of change algorithm for calcium intake by male college students. AB - Using a sample of college-aged men, we evaluated the validity of a staging algorithm for dairy calcium intake that had been previously validated in a sample of college women. College-aged men enrolled at a private university in the United States were randomly selected to participate. The sample consisted of 180 male participants. Self-reported calcium intake was measured using a 26-item food frequency questionnaire. Stages of change classifications were based on a four question algorithm for dairy calcium intake and a three-item self-efficacy questionnaire. Significant differences were found between dairy/calcium intake levels between pre-contemplation, contemplation/preparation, and action/maintenance groups. Results also showed that 46% of participants were in the action/maintenance stage. There was a significant difference in self-efficacy between members of the pre-action and action stages. The study suggests that this stage of change algorithm for dairy calcium intake is valid for use in a male college student population. PMID- 16720129 TI - Best practice methods to apply to measurement of resting metabolic rate in adults: a systematic review. AB - Several factors may alter apparent resting metabolic rate (RMR) during measurement with indirect calorimetry. Likewise, numerous indirect calorimetry measurement protocols have been developed over the years, and the methodology employed could influence test results. As part of a larger project to determine the role of indirect calorimetry in clinical practice, a systematic review of the literature was undertaken to determine the ideal subject condition and test methodology for obtaining reliable measurement of RMR with indirect calorimetry. Food, ethanol, caffeine, and nicotine affect RMR for a variable number of hours after consumption; therefore, intake of these items must be controlled before measurement. Activities of daily living increase metabolic rate, but a short rest (< or =20 minutes) before testing is sufficient for the effect to dissipate. Moderate or vigorous physical activity has a longer carryover effect and therefore must be controlled in the hours before a measurement of RMR is attempted. Limited data were found regarding ideal ambient conditions for RMR testing. Measurement duration of 10 minutes with the first 5 minutes deleted and the remaining 5 minutes having a coefficient of variation <10% gave accurate readings of RMR. Individuals preparing for RMR measurement via indirect calorimetry should refrain from eating, consuming ethanol and nicotine, smoking, and engaging in physical activity for varying times before measurement. The test site should be physically comfortable and the individual should have 10 to 20 minutes to rest before measurement commences. A 10-minute test duration with the first 5 minutes discarded and the remaining 5 minutes having a coefficient of variation of <10% will give an accurate measure of RMR. PMID- 16720131 TI - The effects of dieting on food and nutrient intake of lactating women. AB - The purpose of this report was to identify and evaluate dietary changes in women who were participating in a study on the effects of weight loss in overweight lactating women on the growth of their infants. Women were randomly assigned at 4 weeks postpartum to either restrict energy intake by 500 kcal/day (diet and exercise group) or to maintain usual dietary intake (control group) for 10 weeks. The diet and exercise group significantly decreased fats, sweetened drinks, sweets and desserts, snack foods, and energy intake. Micronutrient intake decreased in the diet and exercise group; however, mean intakes were not significantly different from those of the control group except for calcium and vitamin D. Both groups consumed less than 76% of the Recommended Dietary Allowance for vitamins E and C at the end of the study. Mean intake of all other nutrients was adequate in both groups. These results suggest that overweight lactating women can restrict their energy intake by 500 kcal per day by decreasing consumption of foods high in fat and simple sugars. However, they must be advised to increase their intakes of foods high in calcium and vitamin D. Increased intake of fruits and vegetables should also be recommended to all lactating women, as well as multivitamin and calcium supplements to those who do not consume adequate amounts of these foods. PMID- 16720132 TI - An educational program enhances food label understanding of young adolescents. AB - While much research has focused on intake and eating behaviors of young adolescents, few studies have examined their knowledge of the Nutrition Facts label. The goals of this project were to assess the understanding of Nutrition Facts labels and the response of young adolescents to an educational program about Nutrition Facts labels. Thirty-four young adolescents participated by answering a short questionnaire and taking a pretest about Nutrition Facts labels and their food choices, which involved using actual Nutrition Facts labels from foods typically eaten by adolescents. Then the subjects participated in a short educational program on how to read the Nutrition Facts label. This was followed by a posttest. Initial knowledge of the basic use of the Nutrition Facts label was marginal, with only 6.65 (standard deviation=2.23) or approximately 55% of pretest questions answered correctly. However, the subjects' scores improved significantly to 8.32 (standard deviation=2.01) or approximately 70% correct on the posttest (P<0.0001, Fisher's exact test) after the educational session. In conclusion, young adolescents can learn how to read and understand the Nutrition Facts labels through educational sessions. PMID- 16720133 TI - Consumers may not use or understand calorie labeling in restaurants. AB - This study was an investigation of the possible utility of calorie labeling legislation in restaurants in community (n=649) and college student (n=316) samples. Only 48% to 66% of participants presently looked at food labels, and 64% to 73% were able to report accurate knowledge of daily caloric needs. Furthermore, 44% to 57% reported that they were not likely to use food label information in restaurants if it were available. Therefore, public education campaigns focused on calorie requirements may need to precede restaurant labeling, and perhaps other possibilities in labeling formats should be considered (eg, defining foods as "low," "moderate," and "high" calorie). PMID- 16720134 TI - Experiences of food insecurity among urban soup kitchen consumers: insights for improving nutrition and well-being. AB - Adequate nutrition is an essential determinant of health. Disadvantaged individuals within the cities of developed countries continue to have poor health, yet the role of food insecurity in such groups is poorly understood. This cross-sectional study describes such experiences among 22 randomly selected participants who participated in interviews at a charity-run soup kitchen in urban Sydney, Australia. Interviews explored four constructs of food insecurity (quantitative, qualitative, psychological, and social), identifying related barriers and coping strategies. Reliable access to food was limited. Low income; high rents; poor health; and addictions to cigarettes, alcohol, illicit drugs, and gambling were associated with dependence on charities. Poor dentition and lack of food storage and cooking facilities were important barriers to adequate nutrition. Meals were missed and quantities restricted as a coping strategy. Participants demonstrated adequate knowledge and a desire to eat healthful food. Opportunities for social interaction and trust in soup kitchen staff were important motivators of attendance. Strategies to reduce food insecurity among seriously disadvantaged city dwellers should focus less on education and more on practical solutions, such as accessing affordable healthful food for those without kitchen facilities, improving dentition, and reducing addictions. It is also important to facilitate social networks with trusted support organizations. PMID- 16720135 TI - Entry-level dietetics practice today: results from the 2005 Commission on Dietetic Registration Entry-Level Dietetics Practice audit. PMID- 16720136 TI - Distinguishing entry-level RD and DTR practice: results from the 2005 Commission on Dietetic Registration Entry-Level Dietetics Practice audit. PMID- 16720137 TI - What are the facts and myths about mangosteen? PMID- 16720138 TI - The impact of prophylactic antibiotics on postoperative infection complication in elective laparoscopic cholecystectomy: a prospective randomized study. AB - BACKGROUND: The aim of this prospective randomized study was to investigate the necessity and impact of prophylactic antibiotics on postoperative infection complications in elective laparoscopic cholecystectomy. METHODS: At the time of induction of anesthesia, group A patients (n = 141) received 1 g cefazolin, and group B patients (control; n = 136) received 10 mL isotonic sodium chloride solution. Patients' characteristics and general operative outcomes were compared and analyzed. RESULTS: The overall rate of infection was 1.1% for total 277 patients (0.7% for group A patients and 1.5% for group B patients). No significant difference in infection complications was found between these 2 groups. Also any risk factors contributing to infection complications could not be found. CONCLUSIONS: We do not recommend the use of prophylactic antibiotics in elective laparoscopic cholecystectomy because they will not decrease the already low rate of postoperative infectious complications. PMID- 16720139 TI - Toward zero pancreatic fistula after pancreaticoduodenectomy with pancreaticogastrostomy. AB - BACKGROUND: Postoperative morbidity after pancreaticoduodenectomy has been associated mainly with the development of pancreatic fistula. However, postoperative complications unrelated to pancreatic fistula cannot be disregarded after pancreaticoduodenectomy. The aim of the present study was to investigate the postoperative morbidity in a large series of pancreaticoduodenectomies with pancreaticogastrostomies without pancreatic fistula. METHODS: The present study analyzed the data from 194 consecutive patients undergoing a pancreaticoduodenectomy with a pancreaticogastrostomy between July 1997 and June 2003 in whom no postoperative pancreatic fistula occurred. RESULTS: The overall rate of postoperative morbidity was 33.5%. Specific and general complications occurred in 16% and 17.5% of the patients, respectively. An American Society of Anesthesiologists (ASA) score of 3 and blood transfusion were the only independent factors associated with postoperative morbidity. CONCLUSIONS: Our study found that the overall morbidity after a pancreaticoduodenectomy with a pancreaticogastrostomy still remains high even in the absence of pancreatic fistula and is associated with the preoperative medical condition (ASA score) of the patients and with blood transfusion. PMID- 16720142 TI - Clinical usefulness of 18-fluorodeoxyglucose positron emission tomography in the management of patients with nonpancreatic periampullary neoplasms. AB - BACKGROUND: 18-Fluorodeoxyglucose positron emission tomography (18-FDG PET) has been investigated for the diagnosis and staging of gastrointestinal malignancies including pancreatic adenocarcinoma. The aim of this study was to examine the clinical usefulness of 18-FDG PET in the diagnosis and follow-up evaluation of patients with periampullary neoplasms. METHODS: Twenty-five patients underwent whole-body 18-FDG PET and abdominal computed tomography (CT). Pathologic confirmation was obtained in all patients by surgical resection or biopsy examination. The 18-FDG PET was analyzed visually and semiquantitatively using the standard uptake value (SUV). Positivity was assumed when a focal uptake occurred with an SUV of 2.5 or greater. RESULTS: Between January 1998 and December 2003, 14 ampullary, 7 bile duct, and 4 duodenal tumors were included in the study. PET showed increased focal uptake in 22 patients (88%): 11 of 14 (79%) ampullary tumors, and 100% of bile duct and duodenal tumors. PET showed a focal uptake in 11 of 12 patients without detectable mass at CT scan, and lymph node metastases in 6 patients. An SUV value of 2.7 discriminated adenomas or noninvasive cancers (n = 6) from invasive malignancies (n = 14). Follow-up evaluation including CT scan and PET was performed in 12 patients: PET showed recurrent disease not seen by CT in 4 patients, confirmed CT findings in 6 patients, and showed an unsuspected primary lung cancer in 1 patient and colon cancer in another patient. CONCLUSIONS: 18-FDG PET is very sensitive for detecting periampullary neoplasms. It may be useful to differentiate benign or borderline lesions from invasive tumors when no mass has been identified by traditional imaging. Finally, it is very useful in the follow-up evaluation of resected patients to identify recurrent disease or other malignancies. PMID- 16720141 TI - Management of involved or close resection margins in 120 patients with colorectal liver metastases: edge cryotherapy can achieve long-term survival. AB - BACKGROUND: The liver resection margin is prognostically significant for patients with colorectal liver metastases. Management plans for patients with suboptimal resection margins have not been adequately addressed. This article reports the long-term results of edge cryotherapy in 120 patients with suboptimal resection margins. METHODS: A retrospective analysis of prospectively collected clinical data of 120 patients with suboptimal hepatic resection margins was performed. Morbidity, mortality, recurrence, and survival results were analyzed. RESULTS: The median length follow-up was 30 months (range 1 to 139). The median disease free interval was 19 months (range 2 to 139). Cryosite, remaining liver, and extrahepatic recurrence rates were 10%, 36%, and 47%, respectively. The median survival was 39 months (range 1 to 139), and 1-, 3-, and 5-year survival rates were 89%, 55%, and 36%, respectively. Cryosite and extrahepatic recurrence were independently associated with a reduced survival outcome. COMMENTS: Edge cryotherapy to suboptimal liver resection margins can achieve a lower local recurrence rate and a longer survival advantage. PMID- 16720143 TI - Radiotherapy omission after breast-conserving surgery is associated with reduced breast cancer-specific survival in elderly women with breast cancer. AB - PURPOSE: To evaluate the effect of radiotherapy (RT) omission on survival in older breast cancer patients treated with breast-conserving surgery. METHODS: Data were analyzed for 4836 women aged 50 to 89 with T1-T2, N0-N1, M0 breast cancer. Tumor and treatment factors, relapse rates, and overall survival (OS) and breast cancer-specific survival (BCSS) were compared between women treated with and without RT in 3 age categories: 50 to 64 (n = 2398), 65 to 74 (n = 1665), and > or = 75 years (n = 773). RESULTS: Median follow-up was 7.5 years. Rates of RT omission significantly increased with advancing age (7%, 9%, and 26% in age 50 64, 65-74, and > or = 75 years respectively, P < .0001). RT omission was associated with significantly reduced local control, BCSS, and OS. Despite similar tumor characteristics and higher rates of systemic therapy use, women aged > or = 75 years were observed to have lower 5-year OS and BCSS when RT was omitted. CONCLUSION: These findings support the hypothesis that inadequate local therapy is associated with reduced survival in elderly women treated with breast conserving therapy. PMID- 16720144 TI - Elective neck dissection for carcinomas of the oral cavity: occult metastases, neck recurrences, and adjuvant treatment of pathologically positive necks. AB - BACKGROUND: Supraomohyoid neck dissection (SOHND) is currently performed in patients with carcinoma of the oral cavity with clinically negative neck. Most investigators consider SOHND as a staging procedure. METHODS: Records of 100 patients with cancer of the oral cavity and clinically negative neck undergoing SOHND were reviewed. The rate and significance of occult metastases are evaluated, the neck recurrences are analyzed and the indication of adjuvant radiation of pN+ necks is discussed. RESULTS: In 34 of 1814 of analyzed lymph nodes, metastatic disease was detected as follows: 30 macrometastases and 4 micrometastases. In 13 of 34 metastases (38%), extracapsular spread was observed. Twenty of 100 patients (20%) had to be upstaged. In 9 of 87 (10%) patients without local recurrence and with a minimal follow-up of 24 months, 5 ipsilateral (4 within the dissection field) and 5 contralateral neck recurrences were observed. Regional recurrence developed in 4% and 35% of patients with pN0 and pN+ necks, respectively. CONCLUSIONS: In 20% of patients with oral cavity tumors and pN0 neck, occult metastases were disclosed. Neck recurrences developed significantly more often in patients with pN+ than in those with pN0 necks. To evaluate the exact indication for an adjuvant treatment of patients with cN0/pN+ necks, prospective studies should be performed. PMID- 16720146 TI - Prevention of nausea and vomiting following breast surgery. AB - BACKGROUND: The purpose of this study was to determine the rate of nausea and vomiting in women following breast surgery (PONV) under general anesthesia (GA), before and after the introduction of a standardized prophylactic anti-emetic (AE) regimen. METHODS: We performed a retrospective review of eligible patients, between July 2001 and March 2003. Patients operated on before September 2002 had standard preoperative care (old cohort [OC]); patients operated on after September 2002 were treated prophylactically with oral dronabinol 5 mg and rectal prochlorperazine 25 mg (new cohort [NC]). Data were collected from hospital records regarding age, diagnosis, comorbid conditions, previous anesthesia history, anesthesia and operative details, episodes PONV, and use of AE. The rate and severity of PONV was calculated for both cohorts. RESULTS: Two hundred forty two patients were studied: 127 patients in the OC and 115 patients in the NC. The median age was 56 years (range 32 to 65). The rate of nausea and vomiting were significantly better in the patients treated prophylactically with dronabinol and prochlorperazine (59% vs. 15%, P < .0001 and 29% vs. 3%, P < .0001). Twenty patients in the OC were given some prophylactic AE treatment and 12 (60%) of them required further treatment; only 12 of 109 patients (11%) in the NC required further AE treatment (P < .0001). CONCLUSION: PONV is a significant problem in breast surgical patients. Preoperative treatment with dronabinol and prochlorperazine significantly reduced the number and severity of episodes of PONV. PMID- 16720145 TI - Sentinel lymph node positivity of patients with ductal carcinoma in situ or microinvasive breast cancer. AB - PURPOSE: The purpose of this study was to determine the rates of sentinel lymph node (SLN) positivity in patients with a final diagnosis of ductal carcinoma in situ (DCIS) or microinvasive breast cancer (MIC). METHODS: One hundred thirty patients underwent SLN mapping from 1998 to 2003 for DCIS or MIC. RESULTS: One hundred nine patients with DCIS and 21 with MIC underwent SLN mapping. One patient with bilateral DCIS underwent 2 SLN procedures; therefore, the results of 131 SLN procedures are included. On hematoxylin and eosin (H&E) staining, 4 of 110 patients (3.6%) with DCIS had positive SLNs. Four additional patients had positive SLNs by IHC staining only (3.6%). Two of 8 patients underwent completion axillary dissection, and neither had additional involved nodes on completion axillary dissection. One of the 21 patients with MIC had positive SLNs by hematoxylin and eosin (H&E) (4.8%), and another had an involved SLN by IHC staining (4.8%). The patient with the positive SLN by H&E had 1 additional node on completion axillary dissection. CONCLUSION: Rates of SLN positivity for patients with DCIS are modest, even in a high-risk population, and there is continuing uncertainty about its clinical importance. PMID- 16720148 TI - Open repair of pectus excavatum and carinatum deformities with minimal cartilage resection. AB - BACKGROUND: Numerous modifications of the Ravitch open repair of pectus excavatum (PE) and carinatum (PC) have been used by surgeons with inconsistent results. METHODS: During a 3-year period, 275 consecutive patients underwent open repair of PE and PC using a new less invasive technique. A small chip of costal cartilage was resected medially and laterally from each deformed cartilage, allowing it to barely touch the sternum and rib (laterally) after the sternum had been elevated or depressed, and twisted to the desired position. A support strut used for all patients was routinely removed within 6 months. RESULTS: With mean follow-up of 17 months, all but 5 patients regarded the results as very good or excellent. There were no major complications or deaths. CONCLUSION: Open repair using minimal cartilage resection is effective for all variations of PE and PC in patients of all ages, causes only mild pain, and produces good physiologic and cosmetic results. PMID- 16720147 TI - How accurate are published recurrence rates after rectal prolapse surgery? A meta analysis of individual patient data. AB - BACKGROUND: The literature shows wide variations of recurrence rates (RRs) after abdominal surgery for rectal prolapse. The aim of this meta-analysis was to evaluate the accuracy of published RRs. METHODS: An electronic search was performed with no restrictions. Inclusion criterion was abdominal surgery in at least 10 adults with follow-up evaluation of any length of time. Two reviewers screened 1669 references. A total of 190 investigators were asked to provide individual patient data that should be the same used at the time their reports were written. The RR was estimated by actuarial analysis. Investigators were asked for comments on results. RESULTS: Individual patient data from 6 reports published with 273 patients (186 women, 87 men) with a median age of 54 years (range, 18-88 y) were available. Abdominal surgery included mobilization with pexy (88%), with additional resection (7%), or mobilization only (5%). There were 16 recurrences at a median follow-up period of 3.94 years (range, .05-15.11 y). The effect of age (hazard ratio [HR], 2.010; P = .3443), sex (HR, 2.070; P = .4260), surgical technique (HR, .743; P = .7669), and publication (HR, 1.014; P = .8747) on RR was not significant. Two publications reported a RR of 0. In another report, the published and estimated RRs of 15% did not differ. Published RRs differed from estimated RRs in 3 reports (2.5% vs 4%; 7% vs 54%; and 9.6% vs 36%). The pooled odds ratios of 6 reports revealed a borderline significant difference between the published and estimated RRs (P = .066). CONCLUSIONS: Published RRs differed by as much as 47% from the RRs estimated by actuarial analysis depending on event definition and how the data were censored. PMID- 16720150 TI - Can LigaSure seal and divide the small bowel? AB - BACKGROUND: The LigaSure system (Valleylab, Boulder, CO) seals vessels by reforming collagen and elastin in vessel walls. We studied the feasibility and effectiveness of LigaSure in dividing porcine small bowel. METHODS: Twelve porcine small bowel portions were randomized to division with either endoscopic linear stapler or LigaSure system and the burst pressure of the sealed ends were measured. RESULTS: Segments sealed with staples (6), LigaSure Atlas (Valleylab) (7), LigaSure Xtd (Valleylab) (4), and double-sealed with LigaSure Atlas (Valleylab) (4) were compared. Burst pressure for stapled segments was 131 (standard deviation [SD] 19) mm Hg; failure occurred in the bowel wall. Burst pressures for the 3 groups of Ligasure segments were 27 (SD 5), 20 (SD 6), and 11 (SD 13) mm Hg, respectively, with no statistically significant difference among them; failure occurred at the sealed end. Burst pressure for stapled segments was significantly higher than that of Ligasure segments (P < .001). CONCLUSION: LigaSure does not safely seal small bowel. PMID- 16720149 TI - Modifications in the production of cytokines and growth factors in drainage fluids following mesh implantation after incisional hernia repair. AB - BACKGROUND: The aim of this study was to evaluate changes in the production of some cytokines (interleukins [ILs]-6, -10, -1, and -1ra), vascular endothelial growth factor, and beta-fibroblast growth factor after polypropylene mesh implantation. METHODS: Twenty female patients were divided into 2 groups. In 1 group, hernia repair was performed with conventional sutures (CR), whereas in the other group polypropylene mesh (MR) was used. Growth factors and cytokines production was analyzed in wound drain fluids based on the amount produced during 24 hours. RESULTS: IL-1 increased substantially in MR patients on postoperative days 1 and 2. IL1-ra and IL-10 production was always significantly higher in CR patients. IL-6 production did not show any considerable difference between the 2 groups. Vascular endothelial growth factor production was significantly higher in the MR than the CR group at all time points, whereas beta-fibroblast growth factor production was higher in the MR than the CR group only on postoperative day 1. COMMENTS: Our data suggest that different surgical procedures induce various levels of inflammation and that implantation of prostheses significantly stimulates the inflammatory response. PMID- 16720151 TI - Demonstration of a biliobronchial fistula with a hepatoiminodiacetic acid scan. AB - Biliobronchial fistula (BBF) is a rare complication after hepatic resection. A 68 year-old woman who had undergone a right hepatic trisegmentectomy followed by a hepaticojejunostomy with a Roux-en-Y anastomosis for gallbladder cancer and liver metastasis from colon cancer was admitted to our hospital for further investigation of persistent bilioptysis. We could not detect the cause of the bilioptysis on computed tomography or magnetic resonance cholangiopancreatography, but a hepatoiminodiacetic scan clearly showed the BBF. Endoscopic retrograde cholangiography is thought to be suitable for diagnosing BBF, but it is invasive and unavailable in patients who have undergone digestive reconstruction, such as a Roux-en-Y anastomosis. In such cases, hepatoiminodiacetic scanning should be used for diagnosing BBF. PMID- 16720152 TI - Hemobilia due to cystic artery pseudoaneurysm. PMID- 16720153 TI - Carcinoid tumors in the breast. AB - BACKGROUND: Carcinoid tumors in the breast are rare. Most represent metastases from other primary sites, but commonly are mistaken for primary breast lesions. METHODS: A literature search of the English language found 59 cases of carcinoid tumors in the breast, 21 (36%) of which were metastases. RESULTS: We present an additional 3 cases of carcinoid tumors metastatic to the breast and discuss the clinical, radiologic, and pathologic manifestations. CONCLUSIONS: It is important to differentiate between primary breast carcinoid tumor and metastatic disease to the breast because of differences in treatment. All palpable breast masses and mammographically detected lesions should undergo a biopsy examination. In those patients with a known history of carcinoid tumor, pertinent clinical history, and previous surgical specimens should be reviewed to avoid an unnecessary mastectomy. If there is no history of a prior carcinoid tumor, a thorough work-up to look for an occult primary tumor elsewhere should be performed. PMID- 16720154 TI - The use of virtual patients to teach medical students history taking and communication skills. AB - BACKGROUND: At most institutions, medical students learn communication skills through the use of standardized patients (SPs), but SPs are time and resource expensive. Virtual patients (VPs) may offer several advantages over SPs, but little data exist regarding the use of VPs in teaching communication skills. Therefore, we report our initial efforts to create an interactive virtual clinical scenario of a patient with acute abdominal pain to teach medical students history-taking and communication skills. METHODS: In the virtual scenario, a life-sized VP is projected on the wall of an examination room. Before the virtual encounter, the student reviews patient information on a handheld tablet personal computer, and they are directed to take a history and develop a differential diagnosis. The virtual system includes 2 networked personal computers (PCs), 1 data projector, 2 USB2 Web cameras to track the user's head and hand movement, a tablet PC, and a microphone. The VP is programmed with specific answers and gestures in response to questions asked by students. The VP responses to student questions were developed by reviewing videotapes of students' performances with real SPs. After obtaining informed consent, 20 students underwent voice recognition training followed by a videotaped VP encounter. Immediately after the virtual scenario, students completed a technology and SP questionnaire (Maastricht Simulated Patient Assessment). RESULTS: All participants had prior experience with real SPs. Initially, the VP correctly recognized approximately 60% of the student's questions, and improving the script depth and variability of the VP responses enhanced most incorrect voice recognition. Student comments were favorable particularly related to feedback provided by the virtual instructor. The overall student rating of the virtual experience was 6.47 +/- 1.63 (1 = lowest, 10 = highest) for version 1.0 and 7.22 +/- 1.76 for version 2.0 (4 months later) reflecting enhanced voice recognition and other technological improvements. These overall ratings compare favorably to a 7.47 +/- 1.16 student rating for real SPs. CONCLUSIONS: Despite current technological limitations, virtual clinical scenarios could provide students a controllable, secure, and safe learning environment with the opportunity for extensive repetitive practice with feedback without consequence to a real or SP. PMID- 16720155 TI - Directed postgraduate study result in quantitative improvement in American Board of Surgery In-Training Exam scores. AB - BACKGROUND: The collective study habits of 1 group of residents involved in educationally distinct periods of time in a community-based general surgery residency program were evaluated. METHODS: American Board of Surgery In-Training Exam (ABSITE) score results of 31 residents were calculated during 3 distinctive educational time periods: resident independent, self-directed study; resident directed study with weekly systematic textbook reviews; and faculty-directed study with additional formal basic science and clinical lectures. RESULTS: Aggregate higher scores were observed when ABSITE results for the directed study period were compared with those observed during the independent study period in mid-level resident years (postgraduate year [PGY] 2 to 4). CONCLUSIONS: With limited faculty resources, community-based surgery residency programs have more challenges in opportunities for resident acquisition of cognitive knowledge and subsequent quantitative improvement in ABSITE scores. This study demonstrated a successful methodology particularly in the face of mandated limitation of weekly resident work hours and diminishing allocated education resources. PMID- 16720156 TI - Assessment of basic human performance resources predicts performance of ureteroscopy. AB - BACKGROUND: Our objective was to predict endoscopic performance in a cadaver model using basic performance resources (BPRs) measurements. METHODS: Medical students (n = 16) underwent intense ureteroscopic training on a virtual reality ureteroscopy trainer and were rated on performing ureteroscopy on a cadaver. The medical students also underwent 13 validated BPR measurements. Urology residents also performed cadaveric ureteroscopy and BPRs. A predictive model built from urology residents' (n = 16) BPRs and performance assessment was used to predict medical student cadaveric ureteroscopy performance based on their BPRs alone. RESULTS: The predictive model built with urology residents predicted the ureteroscopic performance of 10 of 16 medical students within 15% of their rated ureteroscopic performance on the cadaver. CONCLUSIONS: A predictive model built with urology residents can moderately predict the ureteroscopic performance of medical students from BPRs. Additional in vivo evaluation is required. PMID- 16720157 TI - Easyloop knot: a simple and safe extracorporeal knot. AB - Thorough knowledge of laparoscopic suturing is of great importance to the laparoscopic surgeon, especially during the performance of advanced laparoscopic procedures. Intracorporeal and extracorporeal knot tying enhances the technical capabilities of the laparoscopic access, thus extending the spectrum of laparoscopic procedures to that of open surgery. We describe herein a new extracorporeal knot designed with an emphasis on simplicity and safety. PMID- 16720158 TI - Inferior branch-preserving superior head resection of the pancreas with gastric wall-covering method for intraductal papillary mucinous adenoma. AB - Intraductal papillary mucinous neoplasm (IPMN) of the pancreas showed significantly less aggressive pathologic features compared with common pancreatic cancer. Therefore, several organ-preserving techniques, including duodenum preserving pancreatic head resection, have been advocated in surgical procedures for IPMN. We describe a surgical technique of superior head resection of the pancreas with inferior branch preservation followed by a gastric wall-covering method for the prevention of pancreatic leakage in patients with IPMN of the pancreas head. PMID- 16720159 TI - Axillary lymph node count is lower after neoadjuvant chemotherapy. AB - BACKGROUND: Retrieval of fewer than 10 lymph nodes at axillary dissection (ALND) for breast cancer can represent anatomic variation or inadequate dissection. We postulated that despite aggressive ALND, a lower lymph node count is more frequent after neoadjuvant chemotherapy. METHODS: Patients who received neoadjuvant chemotherapy followed by ALND were compared with patients who received surgery first. All patients received a level I and II ALND at a single institution by one of the breast surgeons. The number of nodes retrieved at ALND was dichotomized into categories (< 10 and > or = 10), and compared using Fisher exact test. RESULTS: A total of 143 neoadjuvant and 170 surgery-first patients were studied. Patients treated with neoadjuvant chemotherapy were significantly more likely to have fewer than 10 lymph nodes retrieved at ALND than were the surgery-first patients (19/143 or 13% vs. 6/170 or 4%, P = .003). CONCLUSIONS: A low lymph node count is more common in patients after treatment with neoadjuvant chemotherapy and should not be assumed to represent an incomplete ALND. PMID- 16720160 TI - Lower nodal counts in axillary dissection following neoadjuvant chemotherapy: are there implications? PMID- 16720163 TI - C-Met antisense oligodeoxynucleotide inhibits growth of glioma cells. AB - BACKGROUND: C-Met, a receptor tyrosine kinase, and its ligand, hepatocyte growth factor, are critical in cellular proliferation, motility, and invasion and are known to be overexpressed in gliomas. The aim of our study was therefore to investigate the effect of transfected caroboxyfluorescein-5-succimidyl ester (FAM)-labeled c-Met antisense oligonucleotide (ASODN) on growth of glioma cells. METHODS: Conjugated FAM-labeled c-Met ASODN was encapsulated by LIPOFECTAMINE PLUS Reagent and then added into the human glioma cell line U251. Cultured cells were divided into 5 groups: control group, 500 nmol/L nonsense oligonucleotide (NSODN) group, 250 nmol/L ASODN group, 500 nmol/L ASODN group, and 750 nmol/L ASODN group. The intracellular distribution of c-Met ASODN was observed with fluorescence microscopy; cell growth was detected by methyl thiazole tetrazolium assay. The apoptosis of U251 cells was also examined with a flow cytometer. Semiquantitative reverse transcriptase polymerase chain reaction and Western blot examinations were carried for expression of c-Met messenger RNA (mRNA) and protein. RESULTS: The blue fluorescence was seen in the cytoplast and nuclei of cells of FAM-labeled c-Met ASODN groups with fluorescence microscopy after the cells were treated with FAM-labeled c-Met ASODN-LIPOFECTAMINE PLUS Reagent complex for 3 hours. Antisense (AS) oligonucleotide caused a statistically significant reduction of cell viability (P < .05), whereas NSODN had no such changes. The cell growth was also significantly inhibited by ASODN (P < .05). After transfection, 250, 500, and 750 nmol/L ASODN induced significant apoptotic response, about 4.67% +/- 2.86%, 8.65% +/- 3.18%, and 12.76% +/- 3.15% for 24 hours (P < .05) and 7.79% +/- 1.92%, 11.43% +/- 1.54%, and 15.78% +/- 1.86% for 48 hours (P < .01), respectively. However, 500 nmol/L NSODN did not induce any significant apoptotic response until 48 hours after transfection (P > .05). A significant loss of c-Met mRNA was presented in ASODN-treated cells, and this was not seen in treatment with NSODN. Protein level was significantly decreased 48 hours after c-Met ASODN transfected. CONCLUSIONS: Antisense oligonucleotide targeting c-Met can be identified as a most potent AS compound, which can inhibit cell growth and induce cell apoptosis. This provides evidence that c-Met plays a role in tumor progression of glioma by acting as an oncogene and suggests that c Met ASODN may provide a novel approach to therapy for human glioma. PMID- 16720165 TI - Efficacy and safety of hypertonic saline solutions in the treatment of severe head injury. AB - BACKGROUND: The present study was undertaken to evaluate the efficacy and safety of hypertonic saline (HS) in the treatment of intracranial hypertension after severe head injury. METHODS: This prospective, observational study was performed in an 11-bed neurosurgery intensive care unit of a teaching hospital. From February 2002 to September 2004, 18 severely head-injured patients with elevated intracranial pressure (ICP) and Glasgow Coma Scale scores of 5 to 8 (mean, 5.9 +/ 1.2) were admitted to the unit and treated according to a standard protocol. One dose per day of 3% saline was administered by rapid infusion (300 mL/20 min) when ICP values exceeded 20 mm Hg. After infusion, cerebral blood flow, ICP, blood pressure, end-tidal carbon dioxide, and heart rate were monitored continuously for 60 minutes and recorded. Serum osmolarity, sodium, potassium, chloride, arterial carbon dioxide pressure, arterial oxygen pressure, hemoglobin, lactic acid, and pH were measured immediately before infusion (zero time) and 20 and 60 minutes after infusion. Mean arterial pressure, cerebral perfusion pressure (CPP), mean flow velocity (MFV), and pulsatility index (PI) were also recorded and analyzed. RESULTS: Intracranial pressure fell immediately after initiation of infusion with further significant decreases observed at 20 and 60 minutes (30.4 +/- 8.5, 24.3 +/- 7.4, and 23.8 +/- 8.3 mm Hg, respectively; P < .01). At these respective times CPP increased significantly (78.7 +/- 8.7, 83.2 +/- 7.8, and 87.2 +/- 12.8 mm Hg), PI dropped rapidly (1.51 +/- 0.42, 1.38 +/- 0.32, and 1.34 +/- 0.33) and MFV increased (66.26 +/- 25.91, 71.92 +/- 28.13, and 68.74 +/- 28.44). Serum sodium increased from 141.3 +/- 7.2 to 146.3 +/- 7.2 mmol/L after 20 minutes and returned to 144.3 +/- 7.36 mmol/L at 60 minutes. Potassium concentrations decreased significantly from 3.9 +/- 0.39 to 3.55 +/- 0.35 mmol/L after 20 minutes (P < .01). Lactic acid values at 0, 20, and 60 minutes were 1.6 +/- 0.5, 1.47 +/- 0.48, and 1.38 +/- 0.53 mmol/L, respectively (P < .01). CONCLUSION: Rapid infusion of single dose daily of HS is a safe alternative for the treatment of elevated ICP in severe head injury. Further evaluations of long term consequences and complications and of maximal tolerance to this treatment are required. PMID- 16720167 TI - Endoscopic surgery for spontaneous basal ganglia hemorrhage: comparing endoscopic surgery, stereotactic aspiration, and craniotomy in noncomatose patients. AB - BACKGROUND: This prospective study aimed to evaluate the safety, neurological outcomes, and cost-effectiveness of 3 surgical procedures for spontaneous basal ganglia hemorrhage. METHODS: Ninety noncomatose patients with basal ganglia hemorrhages were randomized into 3 groups. Group A (n = 30) underwent endoscopic surgery, group B (n = 30) underwent stereotactic aspiration, and group C (n = 30) underwent craniotomy. Waiting time of surgery, length of operation time, and blood loss were compared between all groups. On the second operative day, we evaluated the amount of residual hematoma and the hematoma evacuation rate by computed tomography scan. Surgical mortality and complications were recorded 3 months after the procedure. Neurological outcomes were evaluated by functional independence measure (FIM) score, Barthel index score, and muscle power (MP) of affected limbs 6 months after surgery. We also evaluated the cost-effectiveness of each procedure. RESULTS: There was significant delay in waiting timing of the stereotactic aspiration (172.56 +/- 93.18 minutes; P < .001). Craniotomy had the longest operation time (229.96 +/- 50.57 minutes; P < .001). Blood loss was most significant in the craniotomy (236.13 +/- 137.45 mL; P < .001). The highest hematoma evacuation rate was seen in the endoscopic surgery (87% +/- 8%; P < .01). The mortality rate was 0% in group A, 6.7% in group B, and 13.3% in group C (P = .21). The complication rate was 3.3% in group A, 10% in group B, and 16.6% in group C (P = .62). The most major complications were rebleeding and infection. The FIM score was higher in the endoscopic surgery (79.90 +/- 36.64) than in the craniotomy (33.84 +/- 18.99; P = .001). The Barthel index score was also significantly better in the endoscopic surgery (50.45 +/- 28.59) than in the craniotomy (16.39 +/- 20.93; P = .006). There was more improvement in MP of affected limbs in endoscopic surgery than in craniotomy (P = .004). Endoscopic surgery was more cost-effective than craniotomy using FIM and Barthel index (P < .02 and P < .05, respectively). CONCLUSIONS: Both endoscopic surgery and stereotactic aspiration are minimally invasive and are effective procedures with low complication and mortality rates; however, the waiting timing of stereotactic aspiration is usually longer. Endoscopic surgery may be an appropriate substitute for stereotactic aspiration. It produces good neurological outcomes and aids in rapid hematoma evacuation. Craniotomy may be used for emergency decompression of enlarged hematoma if endoscopic surgery or stereotactic aspiration is not available. PMID- 16720170 TI - Brain abscess in 142 patients: factors influencing outcome and mortality. AB - BACKGROUND: With the introduction of CT, stereotactic techniques, and broad spectrum antibiotics, the outcome for brain abscess has dramatically improved. The purpose of this study was to identify prognostic factors by reviewing data on 142 patients with brain abscess. METHODS: Clinical data, including age, sex, medical history, duration of symptoms, initial neurological status, associated predisposing factors, laboratory data, treatment, and abscess characteristics, were considered as potential prognostic factors. A comparison was made between patients with favorable (GOS: moderate disability or good recovery) and those with unfavorable (GOS: death, persistent vegetative status, or severe disability) outcomes at discharge. Univariate (chi(2) analysis or Fisher's exact test) and multivariate logistic regression analyses were used to identify prognostic factors. Data were considered significant when the 2-tailed P value was lower than .05. RESULTS: There were 98 male and 44 female patients (male/female ratio, 2.2). Their average age at diagnosis was 41.5 years (range, 2-84 years). There were 105 patients with a favorable outcome and 37 with an unfavorable outcome. Both univariate and multivariate analyses indicated that patients who were male, had an initial GCS score >12, had no other septic complication, or had Gram positive cocci grown in abscess cultures had better outcomes. No association was found between outcome and other factors, including age, focal neurological deficits, seizures, laboratory findings, characteristics of the abscesses, associated factors, and treatment modalities. CONCLUSIONS: With the advancement of imaging studies and broad-spectrum antibiotic therapies, the outcome of brain abscess depends on prompt awareness of the diagnosis and effective infection control. PMID- 16720173 TI - Factors that affect the surgical outcomes of spinal dural arteriovenous fistulas. AB - BACKGROUND: Authors disclose the factors that affect the surgical outcome of the spinal dural AVFs from the retrospective analysis of 13 cases. METHODS: Thirteen patients with spinal dural AVF underwent microsurgical treatments between 1990 and 2004 at the Department of Neurosurgery, Aso Iizuka Hospital. The mean age was 62 years, and the median time to diagnosis was 38 months (range, 4-120 months). There were 12 men and 1 woman. The clinical features were characterized by spastic paraparesis in all 13 patients and micturition problem in 11 patients. All 13 patients showed the longitudinal extension of the high T2 intramedullary signals in magnetic resonance (MR) image. The microsurgical obliterations of the spinal dural AVFs were performed for all 13 patients because the endovascular treatments were difficult or failed to obliterate the lesions. RESULTS: The durations of the symptoms were not directly correlated to the preoperative neurological conditions and the postoperative outcomes. The patient's age and the preoperative severity of myelopathy affected the postoperative outcomes. The mode of the longitudinal extension of the high T2 intramedullary signals in MR image, rostral or caudal, did not show the statistical significance to the outcome. The poor outcomes were observed in elderly patients and patients with preoperative modified Rankin Scale 4 and 5. A case with multiple spinal dural AVFs is also presented. CONCLUSION: The spinal AVFs are treatable lesions through microsurgery or embolization. The likelihood of favorable outcome was affected by the severity of deficits and the patient's age. PMID- 16720176 TI - Intraoperative monitoring of cerebral blood oxygenation and hemodynamics during extracranial-intracranial bypass surgery by a newly developed visible light spectroscopy system. AB - BACKGROUND: Cerebrovascular reconstruction procedures run the risk of changing the balance between oxygen supply and consumption during surgery. We assessed the value of visual light spectroscopy for detecting changes in cerebral blood oxygenation (CBO) during superficial temporal artery-middle cerebral artery (STA MCA) anastomosis. METHODS: We developed a VLS monitoring system which permits continuous monitoring of CBO changes during surgery. Using the VLS, we evaluated the CBO changes in the MCA territory on the lesion side in 18 patients who underwent STA-MCA anastomosis. RESULTS: Temporary occlusion of the MCA (M4 portion) did not change the CBO in 17 patients. However, in the patient with dissecting aneurysm, it caused decreases of oxyhemoglobin and cortical oxygen saturation (CoSo(2)) associated with an increase of deoxyhemoglobin, although these CBO changes were normalized by STA blood flow. In 5 patients, STA blood flow increased the oxyhemoglobin and CoSo(2) and decreased the deoxyhemoglobin, indicating that cortical blood flow (CoBF) was increased. The CoSo(2) before anastomosis was significantly low in the patients who showed an increase of CoSo(2) by STA blood flow (63.0% +/- 2.5%) as compared with those who did not (72.0 +/- 6.1%, P = .024). CONCLUSION: Temporary occlusion of a cortical artery during bypass surgery did not affect the CBO in patients who had chronic cerebral ischemia, but caused acute ischemia in the patient who did not. STA blood flow increased the CoBF during surgery more frequently in patients who showed a low perfusion pressure. The VLS monitoring system is considered useful for evaluating bypass function and facilitates safe and accurate bypass surgery. PMID- 16720178 TI - Cognitive impairment associated with intraoperative and postoperative hypoperfusion without neurologic deficits in a patient undergoing carotid endarterectomy. AB - BACKGROUND: Neuropsychological testing detects cognitive impairment in 20% to 30% of patients after carotid endarterectomy (CEA). CASE DESCRIPTION: A 51-year-old man with asymptomatic right cervical internal carotid artery (ICA) stenosis underwent a CEA. Intraoperative transcranial regional cerebral oxygen saturation monitoring revealed ischemia in the right cerebral hemisphere during ICA clamping and transient hyperemia subsequent to ICA declamping. The patient recovered without the appearance of new neurologic deficits. Brain single-photon emission computed tomography performed immediately after CEA showed a decrease in cerebral blood flow in the right cerebral hemisphere. Diffusion-weighted magnetic resonance imaging showed no new abnormal findings. Positron emission tomography performed 2 months after surgery revealed decreased cerebral metabolic rate of oxygen in the right cerebral hemisphere, and neuropsychological testing demonstrated a decline in performance IQ relative to preoperative levels. CONCLUSIONS: Intraoperative ischemia and postischemic delayed hypoperfusion in CEA can impair cognition even in the absence of development of postoperative neurologic deficit. PMID- 16720180 TI - Decompressive hemicraniectomy for tumefactive demyelination with transtentorial herniation: observation. PMID- 16720181 TI - Proximal nerve root capillary hemangioma presenting as a lung mass with bandlike chest pain: case report and review of literature. AB - BACKGROUND: Capillary hemangiomas occasionally occur in the peripheral nervous system (PNS), presenting as mass lesions causing proximal nerve root symptoms, thus prompting neurosurgical intervention. In contrast to cavernous angiomas, which may also occasionally involve the PNS, capillary hemangiomas have little or no tendency for acute or chronic bleeding and, hence, lack the apoplectic symptomatic onset and neuroimaging features of hemosiderin deposition as seen in cavernous angiomas. CASE DESCRIPTION: A 56-year-old man presented with chronic, radiating, bandlike, chest wall pain. Chest radiograph revealed an apical lung mass suspicious for malignancy. Subsequent magnetic resonance imaging revealed an enhancing dumbbell-shaped mass extending into the spinal canal and chest cavity at T3, causing a slight degree of lung compression. Differential diagnostic considerations included schwannoma, neurofibroma, and meningioma. At operation, the spongy, reddish purple vascular tumor was easily separated from dura but had to be excised en bloc, with the T3 nerve root at the neural foramen. The large intrathoracic portion measuring 37 x 41 mm was left behind. Pathology was consistent with a capillary hemangioma. CONCLUSIONS: We present the second case in the English language literature of a dumbbell-shaped, epidural capillary hemangioma with intrathoracic extension. We review the literature on proximal nerve capillary hemangiomas and contrast these with cavernous angiomas and capillary hemangioblastomas, both of which occasionally involve proximal nerves. Unlike capillary hemangioblastomas, which may be multiple and associated with von Hippel-Lindau (VHL) syndrome, capillary hemangiomas are solitary lesions and have not been associated with an inherited disorder. PMID- 16720183 TI - Secondary central nervous system involvement by follicular lymphoma: case report and review of the literature. AB - BACKGROUND: We report a patient with indolent stage IV follicular lymphoma, grade 1, initially successfully treated with chemotherapy, who later developed aggressive diffuse large B-cell lymphoma in the parieto-occipital lobe 8 years after initial presentation. The differing patterns of lymphomatous involvement of the central nervous system (CNS) are briefly reviewed, with a focus on the patterns seen in secondary CNS spread by low-grade lymphomas. CASE DESCRIPTION: A 53-year-old man was diagnosed with stage IV follicular lymphoma, grade 1, in 1996. Although initial chemotherapy was successful, he developed several recurrences of lymphoma over the following years. In May 2004, he presented with a discrete, single, massive parieto-occipital lobe brain lesion. The mass failed to regress with empiric cranial external beam radiotherapy. Because of suspicion of an unusual infection, the lesion was surgically excised in its entirety. The mass proved to be an aggressive diffuse large B-cell lymphoma, transformed from his previous follicular cell lymphoma, with retention of strong Bcl-2 and Bcl-6 immunoreactivity. CONCLUSIONS: Parenchymal brain involvement, as opposed to dural or leptomeningeal, is a relatively uncommon pattern of spread to the CNS for systemic lymphomas. More significantly, follicular lymphomas are one of the least frequent types of indolent lymphomas to develop clinically apparent, secondary CNS spread. The presentation of an indolent follicular lymphoma with transformation to an aggressive diffuse large B-cell lymphoma within the brain parenchyma is rare. Its manifestation as a massive, singular lesion is unique and prompted diagnostic confusion. PMID- 16720184 TI - Meningioangiomatosis associated with neurofibromatosis: report of 2 cases in a single family and review of the literature. AB - BACKGROUND: Meningioangiomatosis (MA) is a rare benign disorder. It may occur sporadically or in association with neurofibromatosis (NF). The sporadic type typically presents with seizures, whereas that associated with NF is often asymptomatic. Of the 100 cases reported, only 14 are associated with NF. We now report 2 additional cases of MA associated with neurofibromatosis 2 (NF2) in a single family, with one occurring in the cerebellum. The etiology, pathology, and imaging features of MA are presented. CASE DESCRIPTION: A 38-year-old woman (patient 1) presented with a 4-month history of ataxia. She had been diagnosed previously with NF2. Magnetic resonance imaging (MRI) scans of the brain revealed bilateral acoustic neuromas and multiple calcified intracranial lesions. Her 13 year-old daughter (patient 2) presented with complex partial seizures. MRI scans of the brain revealed bilateral acoustic neuromas and a right parietal mass. Patient 1 underwent a suboccipital craniotomy to resect the right-sided acoustic neuroma. A small portion of normal-appearing cerebellar cortex was resected to avoid undue retraction. Histopathologic examination showed the presence of a lesion consistent with MA. Patient 2 underwent a right temporal-parietal craniotomy to remove the enhancing epileptogenic right posterior temporoparietal lesion. Histopathologic analysis showed a lesion consistent with meningioma and MA. CONCLUSIONS: MA has been reported infrequently in association with NF2. We now report 2 cases of MA associated with NF2 in one family, and we add the cerebellum to possible locations of occurrence. MA should be considered in the differential diagnosis of cortical lesions, particularly in patients with NF2. PMID- 16720185 TI - Psammomatous choroid plexus papilloma: three cases with atypical characteristics. AB - BACKGROUND: Intravertricular papillary neoplasms are derived from choroid plexus epithelium. Although choroid plexus tumors account for 0.4% to 0.6% of all brain tumors, they represent 2% to 4%. Approximately 80% of choroid plexus carcinomas arise in children. CASES DESCRIPTION: We describe 3 cases of choroid plexus papilloma (CPP) with profuse psammomatous bodies and calcifications that have lost their normal papillary architecture. Immunohistochemistry was positive for glial fibrillary acidic protein in 2 cases, and proliferating cellular nuclear antigen index was higher compared with regular CPPs. All 3 patients were female and were 12, 40, and 48 years old, respectively. CONCLUSION: We describe psammomatous CPPs and suggest a difference from CPPs because of the more aggressive clinical course, and higher nuclear proliferation index (proliferating cellular nuclear antigen) than the CPPs that lack psammoma bodies. PMID- 16720186 TI - A large arachnoid cyst of the lateral ventricle extending from the supracerebellar cistern--case report. AB - BACKGROUND: The pathogenetic mechanism of intraventricular arachnoid cyst development is still controversial, but is believed to originate from the vascular mesenchyme or as an extension of the arachnoid cyst in the subarachnoid space into the ventricle through the choroidal fissure. We report a case supporting the extension hypothesis and suggest differential points between an intraventricular arachnoid cyst that extended from the supracerebellar space and a lateral ventricular diverticulum that extended into the supracerebellar cistern. CASE DESCRIPTION: A 12-month-old girl presented with macrocephaly and developmental delay. Her magnetic resonance imaging showed an arachnoid cyst that had developed from the supracerebellar space in the posterior fossa, and which extended into the left lateral ventricle resulting in expansion of the left lateral ventricle and displacing the choroids plexus anteriorly and laterally and the midline to the right. We treated an intraventricular arachnoid cyst by endoscopic fenestration resulting in dramatic reduction of the intraventricular arachnoid cyst with large bilateral subdural fluid collection. We performed a subduroperitoneal shunt for subdural fluid collection and subsequent cystoperitoneal shunt for the remnant cyst. CONCLUSION: We suggest that this case supports the extension hypothesis from the subarachnoid space through the choroidal fissure into the lateral ventricle. We also suggest that one of the radiological differential points between an intraventricular arachnoid cyst and a ventricular diverticulum is displacement and compression of the choroid plexus of the lateral ventricle. PMID- 16720187 TI - Bilateral segmental absence of the internal carotid artery with rete compensation associated with absence of basilar artery: case report. AB - BACKGROUND: Bilateral ICA absence is a rare lesion. Collateral circulation to the middle and anterior cerebral arteries in the absence of one or both ICAs may develop transcranial anastomoses from the external carotid system, a so-called CRM. Very rarely, theses arterial channels are observed in humans. CASE DESCRIPTION: In the following case study, a 38-year-old man presented himself. He had a headache and scalp laceration after having had an accident. A right external carotid angiography identified anastomoses to distal intracranial vessel through the internal maxillary artery, but the intracranial vessels were only faintly visible. A right ascending pharyngeal arteriography showed an abnormal course and anastomoses with the intracranial vessel. The right ICA was absent. The left external carotid angiography demonstrated a network of tortous arteries in the region of the cavernous and petrous portion of ICA. The plexus of vessels on the left side communicated with the proximal part of the cavernous segment of the ICA. Both vertebral angiographies demonstrated a sudden diminution in caliber at the level of vertebrobasilar junction. There was an anomalous arterial collateral circulation originating from vertebral muscular and meningeal branch. There was also an absence of the basilar artery (BA). CONCLUSION: This is the first case with CRM associated with bilateral segmental ICA and BA absence. The cause of this bilateral segmental ICA and BA absence may be maldevelopment of vessel connective tissue or extracellular matrix. PMID- 16720190 TI - Treatment of ruptured fusiform aneurysm in the posterior cerebral artery with posterior cerebral artery-superior cerebellar artery anastomosis combined with parent artery occlusion: case report. AB - BACKGROUND: Surgical management of a fusiform aneurysm in the PCA sometimes requires occlusion of the parent artery. In such cases, reconstruction of the PCA may prevent the development of postoperative visual field defects. CASE DESCRIPTION: A 30-year-old woman with a fusiform aneurysm in the P2 of the left PCA presented with subarachnoid hemorrhage. The left superficial temporal artery and OA were hypoplastic. Left temporal craniotomy revealed that the lateromesencephalic ambient segment of the SCA and the posterior half portion of the P2 segment ran parallel and near each other, and that the calibers of both arteries were relatively consistent. Thus, the 2 arteries were anastomosed in a side-to-side fashion, and the anterior half of the left P2 segment was occluded by clipping proximal and distal to the aneurysm that was located at the left P2 segment just distal to the junction to the posterior communicating artery. The patient had an uneventful postoperative course, and postoperative cerebral angiography demonstrated resolution of the aneurysm, with perfusion of the left PCA from the left SCA via the anastomosis. CONCLUSIONS: Posterior cerebral artery superior cerebellar artery anastomosis is a useful method of achieving PCA reconstruction in the context of a PCA aneurysm requiring occlusion of the parent vessel and when superficial temporal artery or OA-PCA anastomosis cannot be performed. PMID- 16720191 TI - Rotational vertebral artery insufficiency resulting from cervical spondylosis: case report and review of the literature. AB - BACKGROUND: Four cases of vertebrobasilar insufficiency secondary to osteophyte formation at C5-C6 have been reported in the literature. In this article, we report the fifth such case and discuss the utility of dynamic computed tomographic angiography (CTA) in the management of this disorder. CASE DESCRIPTION: A 55-year-old right-handed man presented for evaluation after syncopal episodes associated with right-head turning. Workup revealed cervical spondylosis with stenosis. The C5-C6 level was significantly affected. Dynamic angiography revealed obstruction of vertebral artery flow with right-head turning secondary to an osteophyte at the foramen transversarium at C5-C6. This patient underwent a C5-C6 anterior cervical discectomy and fusion. He also underwent unroofing of the vertebral artery and drilling of the osteophyte at the foramen transversarium. Postoperative CTAs reveal reconstitution of flow in the vertebral artery with head turning. CONCLUSION: The utility of dynamic 3-dimensional CTA in the management of this disorder avoids the risk of invasive studies. PMID- 16720192 TI - Aneurysmal embolization of a blisterlike aneurysm of the internal carotid artery: a case report and review of the literature. AB - BACKGROUND: Little is known regarding the optimal management of a ruptured blisterlike aneurysm of the ICA. Because of the high risk for intraoperative bleeding, direct surgical treatments of these fragile lesions have generally been associated with a poor outcome. We herein report a very rare case of a ruptured blisterlike aneurysm that was successfully treated with coil embolization in the late period. CASE DESCRIPTION: The patient was 21 years old when he had a Hunt and Hess grade IV subarachnoid hemorrhage. At the time of the hemorrhage, 3D-CTA demonstrated a minimal aneurysmal enlargement located in the left C2 portion of the ICA. Because of his poor neurological condition and the risk for a premature rupture during early surgery, delayed surgery was thus scheduled. Cerebral angiography, 13 days later, revealed the shape and size of the aneurysm to have changed in form from a blisterlike aneurysm to a saccular-type one. Initially, we planned to treat the aneurysm by trapping with bypass surgery on the 15th day. However, we instead performed coil embolization on the 19th day because a thick thrombus was found to cover the aneurysm at the time of surgery on the 15th day. CONCLUSION: This is the first report of a ruptured blisterlike aneurysm that was successfully treated with coil embolization in the late period of a subarachnoid hemorrhage after operative confirmation of thrombus formation around the aneurysm. Our findings suggest that coil embolization in the late period appears to be an effective option in the management of selective cases of ruptured blisterlike aneurysms. PMID- 16720194 TI - Surgical exposure in retrosigmoid approach: do we need cerebellar retractors? AB - BACKGROUND: The aim of this study was to present our technique of posterior fossa surgery and to evaluate whether cerebellar retractors are needed. METHODS: Since March 1989, more than 500 surgeries were performed through retrosigmoid approach, and our surgical technique was described. The data were evaluated whether or not cerebellar retractors were used in the operations. RESULTS: The surgeries were vestibular neurectomy, acoustic neuroma removal, and neurovascular decompression in 351, 135, and 14 patients, respectively. Cerebellar retractors have not been used in vestibular neurectomies and in most acoustic neuroma surgeries. The cerebellar retractors have been used in all of the neurovascular decompression surgeries where better exposure of the root entry zones of the cranial nerves is needed, and intracranial endoscopy is applied for further exposure and decompression of the vascular loop. CONCLUSION: There is no need for cerebellar retractors when adequate conditions such as patient positioning, adequate anesthesia techniques, and surgical manipulations are applied in posterior fossa approach except for neurovascular decompression and some of the acoustic neuroma surgeries. PMID- 16720196 TI - A revolution in skull base surgery: the quality of life matters! PMID- 16720199 TI - Novel therapeutic strategies with alemtuzumab for chronic lymphocytic leukemia. AB - In the last 10 years purine analogs have become the chemotherapy of choice for the first-line treatment of chronic lymphocytic leukemia, principally because of their superior efficacy compared with alkylating agents. However, many patients experience a relapse after an initial response or become refractory to these agents. The introduction of immunotherapeutic agents has provided renewed hope for fludarabine-refractory patients. Several clinical trials have shown the efficacy of alemtuzumab in patients with fludarabine-refractory chronic lymphocytic leukemia, including those with poor prognostic factors. Current studies indicate that treatment with alemtuzumab can achieve remissions with undetectable residual disease, as assessed by highly sensitive methods such as quantitative polymerase chain reaction or 4-color flow cytometry. These results suggest new applications for alemtuzumab such as combination treatment with chemotherapeutics or immunotherapeutics, maintenance therapy, and in vivo bone marrow purging prior to transplantation. A number of clinical trials are under way assessing the role of alemtuzumab in these settings. PMID- 16720198 TI - Monoclonal antibody therapy for B-cell malignancies. AB - For decades, chemotherapy was the only available approach for patients with advanced lymphoid malignancies. Treatment paradigms were dramatically altered by the availability of novel and active targeted agents, particularly the monoclonal antibodies, alemtuzumab and rituximab. These agents are now playing an increasingly important role in the treatment of lymphoid malignancies. Alemtuzumab is being used earlier in the course of chronic lymphocytic leukemia in patients with a more intact immune system, when it is likely to have its greatest activity. The immunosuppressive properties of monoclonal antibodies are also being explored in the stem cell transplant setting, including in vivo purging and, with alemtuzumab, for the management of graft-versus-host disease. Rituximab has become ubiquitous in the treatment of most B-cell malignancies. Further research with this antibody is focused on optimizing its use and determining its role in each of the relevant disease states. In addition, newer antibodies are in development for treating chronic lymphocytic leukemia and other B-cell malignancies. New treatment regimens, including combinations of monoclonal antibodies, could enhance complete response rates and prolong progression-free survival, perhaps eventually improving our ability to cure patients with lymphoid malignancies. PMID- 16720200 TI - Beyond detectable minimal residual disease in chronic lymphocytic leukemia. AB - Lymphoproliferative disorders are characterized by the abnormal accumulation of aberrant lymphocytes, which frequently interfere with the processes associated with immunologic response and hematopoiesis. Chronic lymphocytic leukemia (CLL) has traditionally been considered indolent, with a prolonged clinical course. However, a large proportion of patients with CLL have severe symptoms, a poor prognosis, and often require more immediate treatment of their leukemia. Over the last 5 years, technology has transformed the approach to treating patients with CLL. Molecular markers are now available that characterize patients with poor risk disease who may benefit from earlier or more aggressive therapeutic intervention. Biological markers have also been identified that assist in predicting responses to specific agents and may help select an effective therapeutic approach. The advent of more targeted monoclonal antibody therapies, used in combination with chemotherapy regimens or as monotherapy, has the potential to eradicate disease to a point of undetectability by the most sensitive tests available, thereby possibly extending the goal of therapy to include a cure. Because recent data have shown that achieving minimal residual disease (MRD) in the bone marrow is one of the most important factors in predicting duration of remission, MRD may be an appropriate therapeutic end point for patients with poor-risk CLL. The use of rituximab with the cytotoxic agents cyclophosphamide and fludarabine has achieved complete remission with no detectable CLL, as assessed by MRD techniques in a significant proportion of previously untreated and previously treated CLL patients. Monotherapy with alemtuzumab has also been shown to achieve a complete response with undetectable MRD in patients with relapsed/refractory disease. This article reviews recent advances in identifying patients who have poor-risk disease and explores the potential importance of MRD status as an outcome measure of CLL therapy. PMID- 16720201 TI - Strategies in the management of alemtuzumab-related side effects. AB - B-cell chronic lymphocytic leukemia has traditionally been treated with alkylating agents and purine analogues. The introduction of alemtuzumab, a CD52 monoclonal antibody with significant clinical activity in chemotherapy refractory B-cell chronic lymphocytic leukemia, is accompanied by a side effect profile different from that resulting from chemotherapy. The intravenous administration of alemtuzumab is usually accompanied by transient infusion-related side effects manifesting primarily as flu-like symptoms. These reactions can be reduced by use of corticosteroid prophylaxis, and will subside gradually during continued treatment. Alternatively, administration of alemtuzumab subcutaneously may markedly reduce the occurrence of general side effects but results in limited transient local skin reactions in most patients. Neutropenia (grade 4) may occur in approximately 20% of patients, but is usually transient and/or responds promptly to colony stimulating factor therapy; episodes of febrile neutropenia are infrequent. The major side effect of alemtuzumab is T-cell depletion, leading to an increased risk of infection, in particular reactivation of cytomegalovirus. This event typically occurs 3 to 8 weeks after initiation of therapy, coinciding with the T-cell nadir. With vigilance and early detection, these infections are usually manageable and do not cause organ failure. Preliminary data indicate that other infections following alemtuzumab therapy do not seem to occur at a frequency that is higher than expected, probably because of the general prophylactic use of cotrimoxazole (trimethoprim and sulfamethoxazole) and valacyclovir in combination with clinical tumor regressions. The overall safety profile of alemtuzumab appears to be beneficial in relation to disease severity and prognosis in patients with fludarabine-refractory B-cell chronic lymphocytic leukemia. PMID- 16720202 TI - The role of alemtuzumab in nonmyeloablative hematopoietic transplantation. AB - Nonmyeloablative stem cell transplants provide a viable therapeutic option for older patients or patients with comorbid conditions, who were previously deemed to be ineligible for transplantation. Despite improvements in clinical outcomes, graft-versus-host disease (GVHD) remains a significant and potentially lethal complication. One approach by which GVHD has been managed is through introduction of new agents, such as alemtuzumab, into the conditioning regimen. Alemtuzumab is a humanized monoclonal antibody that targets the CD52 antigen, which is highly expressed on both B and T lymphocytes. By depleting T cells in both the donor and the recipient, alemtuzumab has been shown to prevent development of both acute and chronic GVHD, without inhibiting the benefits associated with the graft versus-leukemia effect. Clinical trials have shown that alemtuzumab is able to decrease the incidence of acute and chronic GVHD without impairing engraftment. Furthermore, in chronic lymphocytic leukemia therapy, alemtuzumab has been shown to purge malignant cells from the host to allow for harvesting for the purpose of autologous transplantation. Despite results showing that alemtuzumab can play an important role in managing GVHD, little information is available regarding a standardized dosing schedule. Greater insight into alemtuzumab's pharmacokinetic activity would assist in developing a schedule that can optimize alemtuzumab mediated T-cell depletion to prevent GVHD, while retaining sufficient host T-cell activity to encourage the graft-versus-leukemia effect and prevent relapse. PMID- 16720203 TI - The role of alemtuzumab in the management of T-cell malignancies. AB - T-cell malignancies are rare, making up 10% to 15% of all lymphoid neoplasms in adults. They include many different types of disorders such as T-cell prolymphocytic leukemia, T-cell large granular lymphocytic leukemia, adult T-cell leukemia/lymphoma, cutaneous T-cell lymphoma, and peripheral T-cell lymphoma, which are themselves divided into multiple subcategories. Most T-cell malignancies arise as a result of chromosomal abnormalities, including T-cell receptor rearrangement anomalies. Viral infections are implicated in the development of adult T-cell leukemia/lymphoma and some cases of peripheral T-cell lymphoma have been linked to Epstein-Barr virus or human immunodeficiency virus infection. With the possible exception of T-cell large granular lymphocytic leukemia, which often has an indolent course, T-cell malignancies have not responded well to conventional chemotherapeutic treatment. The introduction of monoclonal antibodies for the treatment of cancer has changed the outlook for patients with T-cell malignancies. Recent studies with single-agent alemtuzumab, an anti-CD52 monoclonal antibody, have shown improved response rates and survival in patients with T-cell prolymphocytic leukemia and cutaneous T-cell lymphoma. Preliminary data also suggest that alemtuzumab may have activity in patients with heavily pretreated peripheral T-cell lymphoma who are refractory to conventional chemotherapy. Preclinical studies with mice bearing human adult T-cell leukemia/lymphoma cells suggest that alemtuzumab may have a potential therapeutic role in this setting. Treatment of T-cell hematologic malignancies with alemtuzumab appears promising. Earlier treatment and combination with chemotherapeutic agents may improve treatment outcome for patients with these malignancies and allow for consolidation with stem cell transplant strategies in selected patients. PMID- 16720204 TI - Mannose-binding lectin: clinical implications for infection, transplantation, and autoimmunity. AB - Mannose-binding lectin (MBL) is a recognition molecule of the lectin pathway of complement and a key component of innate immunity. MBL variant alleles have been described in the coding region of the MBL gene, which are associated with low MBL serum concentration and impaired MBL structure and function. Both high and low serum levels of functional MBL have been associated with a variety of diseases and disease complications. Functioning as double-edged sword, low MBL serum levels have been shown to enhance the risk for infections. On the other hand, high MBL serum levels and high MBL activity have been associated with inflammatory diseases, transplant rejection, and diabetic nephropathy. Underscoring the Jekyll-and-Hyde character of MBL, both high and low serum MBL levels are associated with several aspects of autoimmune diseases. This review provides a general outline of the genetic and molecular characteristics of MBL and discusses MBL-disease association and its consequence in infection, transplantation, and autoimmunity. PMID- 16720205 TI - Soluble HLA-G in heart transplantation: their relationship to rejection episodes and immunosuppressive therapy. AB - The aims of this study were to quantify the level of soluble HLA-G in heart transplant patients, to determine the relationship between the sHLA-G levels and the appearance of acute rejection episodes, and to identify the influence of immunosuppressive therapy on sHLA-G levels. Analysis of sHLA-G, measured by enzyme-linked immunosorbent assay in the transplant patients, revealed the existence of two similarly sized groups of patients. One group displayed a significant increase (p < 0.001) in sHLA-G during the first month after transplantation while the other group maintained low levels of the molecule (0-30 ng/ml) throughout the study. The latter group displayed a high incidence of recurrent severe rejection. A significant increase (p < 0.01) in sHLA-G 2 hours after administration of immunosuppressive treatment (mycophenolate mofetil, cyclosporine A/FK506, corticoids) was found. These results suggest that sHLA-G participates in the induction of certain levels of immunological tolerance in these recipients. PMID- 16720206 TI - Mechanisms of antibody immunotherapy on clonal islet reactive T cells. AB - Clinical intervention trials evaluating the efficacy of antibody immunotherapy in type 1 diabetes are in progress. We tested effects on prediabetic islet antigen specific autoreactive T cells of antithymocyte globulin (ATG) and humanized monoclonal antibodies against CD3 (ChAglyCD3) or CD25 (daclizumab) with regard to downmodulation of the target protein, proliferation, cytokine production, complement-dependent cytotoxicity (CDC), antibody-dependent cell cytotoxicity (ADCC), and survival. ATG leads to depletion of autoreactive CD4+ T cells by ADCC, CDC, and apoptosis, whereas anti-CD3 and anti-CD25 inhibited T-cell autoreactivity in a nondepleting fashion. ATG treatment led to a cytokine burst of Th1- and Th2-associated cytokines. Modulation of cytokine release through humanized monoclonal antibodies was moderate and selective: anti-CD25 led to increased release of IL-2 and reduced production of TNFalpha, whereas anti-CD3 decreased release of interferon-y and IL-5 and increased secretion of IL-10. ATG and the humanized monoclonal antibodies displayed contrasting mechanisms of action. PMID- 16720207 TI - C3d and the septal microvasculature as a predictor of chronic lung allograft dysfunction. AB - Studies have shown a potential role for humoral rejection in the evolution of lung graft dysfunction, apparently based on antibodies without human leukocyte antigen specificity. The correlation between extent of C4d deposition with clinical status further illustrates the importance of humoral immunity. Our study examines the potential value of C3d as a further diagnostic adjunct. C3d deposition was examined in lung allograft specimens using frozen tissue indirect direct immunofluorescence (IIF) and avidin biotin immunohistochemical applied to paraffin embedded tissue. Intermediate/extensive amounts of C3d using IIF and immunohistochemical (IH) methodologies correlated with chronic graft dysfunction; IIF C3d deposition was associated with septal and bronchial wall fibrosis (p < 0.0001). Weak/absent amounts of IIF and IH C3d correlated with clinical stability (p < 0.0001). Higher levels of C3d by IH were more sensitive than by IIF as a bronchiolitis obliterans syndrome determinant. C3d and C4d deposition using immunofluorescence and IH were correlated (p < 0.00001). C3d deposition appears prognostically significant. Higher tissue expression of C3d mark chronic graft dysfunction/persistent graft failure following transplantation. The close correlation between C3d and C4d lends credence to the role of humoral allograft rejection as a pulmonary graft dysfunction contributing factor. C3d by IH manifests higher sensitivity but similar specificity compared to C3d by IIF. PMID- 16720208 TI - Idiopathic pulmonary fibrosis related to endothelial injury and antiendothelial cell antibodies. AB - Mechanisms underlying idiopathic pulmonary fibrosis are not well understood. This paper presents data supporting the hypothesis that microvascular endothelial cell injury and antiendothelial cell antibodies play roles in human idiopathic pulmonary fibrosis. Serologic and pathologic features of 40 patients diagnosed with idiopathic pulmonary fibrosis were evaluated. All patients had open lung biopsies indicating either usual or nonspecific interstitial pneumonitis. All biopsies had morphologic evidence of microvascular injury to the endothelium, and direct immunofluorescence testing revealed variable deposition of IgG, IgM, or IgA within septal microvasculature suggestive of humorally mediated microvascular injury. Ultrastructural studies revealed changes of endothelial cell injury and necrosis and evidence of repetitive episodes of microvascular injury characterized by basement membrane zone collagen deposition and lamellation. Serum samples demonstrated reactivity to multiple endothelial cell antigenic epitopes, and indirect immunofluorescent testing demonstrated a prominent pattern of fluorescence in pulmonary endothelial cell preparations. Serum samples were positive in 37/40 patients for antiphospholipid antibodies with one fourth having positive lupus anticoagulant tests accompanied by thrombotic episodes. In patients with idiopathic pulmonary fibrosis, Factor VIII levels and C-reactive protein levels were also elevated, supporting the presence of endothelial cell injury and inflammation. These data underscore a potential role for immune-based microvascular injury in the evolution of usual or nonspecific interstitial pneumonitis and indicate that those patients have evidence of microvascular injury and endothelial cell necrosis. The high prevalence of antiphospholipid antibodies in these patients may lead to an inherent thrombophilic tendency. PMID- 16720209 TI - Detection of CD8+ T cells sensitized to BK virus large T antigen in healthy volunteers and kidney transplant recipients. AB - BK virus (BKV) infections after renal transplantation are increasingly recognized. Development of immune monitoring strategies against BKV requires definition of antigenic epitopes. Hence, T cells from HLA-A02-positive healthy subjects and kidney transplant recipients were stimulated by BKV lysate pulsed on mature autologous dendritic cells and screened against four different T antigen peptides or against BKV lysate. IFN-gamma production was measured by ELISPOT assays. The peptide BKV362-371 (MLTERFNHIL) was naturally processed and recognized by five of six healthy subjects (39 +/- 11 IFN-gamma spots/100,000 cells) and five of seven kidney transplant recipients (21 +/- 12 IFN-gamma spots). Less frequent and weaker CD8+ T-cell responses were detected against three other peptides. Thus, BKV large T antigen is a target for CD8+ T-cell immunity. T-antigen-specific T-cytotoxic cells circulate in healthy blood donors, implying that transient expression of T antigen presumably occurs at sites of viral latency and helps maintain a constant pool of circulating CD8+ T memory cells. PMID- 16720210 TI - The relationship between HLA class II polymorphisms and somatic deletions in testicular B cell lymphomas of Dutch patients. AB - Several risk factors including immune deficiencies, infections, and autoimmune diseases have been established for non-Hodgkin's lymphoma (NHL). For diffuse large B cell lymphoma (DLBCL), the most common type of lymphoma, no risk factors have been described, which may be due to the intrinsic heterogeneity of this disorder. Previously we reported that, in contrast to nodal DLBCLs, the majority of testicular DLBCLs manifested complete loss of HLA-DR and -DQ expression associated with homozygous deletions of the corresponding genes. To determine the correlation between HLA class II polymorphisms and these lymphomas, we applied DNA typing for HLA-DRB1 and HLA-DQB1 on 50 Dutch patients with testicular and 48 with nodal DLBCL and compared the frequencies with a cohort of healthy Dutch controls. Both the patients with nodal and those with testicular DLBCL manifested significantly higher frequencies of HLA-DRB1*15 than the controls (p < 0.018, odds ratio 2.09 and p < 0.013, odds ratio 2.12, respectively). Moreover, a positive association was seen with HLA-DRB1*12 (p = 0.043, odds ratio 4.17) in the patients with testicular DLBCL, and a negative association was seen with HLA DRB1*07 (p = 0.022, odds ratio 0.13) in the patients with nodal DLBCL. Homozygous deletions of the HLA-DR/DQ region, evaluated by interphase fluorescence in situ hybridization were seen in 20 of 48 testicular tumors. No preferential loss or retention of a particular HLA-DR or -DQ allele was seen because all alleles were at least once retained or involved in a homozygous deletion. PMID- 16720211 TI - Lack of association of IL-10 promoter gene variants with type 1 diabetes in a French population. AB - Although genetic predisposition to type 1 diabetes shows a strong association with human leukocyte antigen (HLA) class II alleles, additional genes may influence the immune process and the progression of beta cell loss. Preliminary reports suggested that IL-10 gene polymorphisms contribute to susceptibility to type 1 diabetes. We analyzed the frequencies of three main variants of the promoter region of the IL-10 gene at the positions -1082, -819, and -592 in a cohort of 358 type 1 diabetic patients representing the same regional population pattern and 519 controls from the same region using an enzyme-linked oligonucleotide sorbent assay. We did not find any statistical association in the entire cohort or after stratification for high-risk HLA-DQ alleles. However, the IL-10 -1082 polymorphism was significantly associated with GAD and IA-2 antibodies at clinical onset. Such polymorphism is known to be associated with the reduction of secreted IL-10 which may support the concept of accelerated Th-1 T-cell reactivity. In conclusion, IL-10 promoter gene variants may contribute, but to a minor extent, to disease susceptibility in juvenile type 1 diabetes and should not be included in the routine genetic screening of high-risk individuals. PMID- 16720212 TI - Association of an extended haplotype of HLA class I alleles and their flanking microsatellites with spondyloarthropathies in South Indian patients. AB - Spondyloarthropathy (SpA) is a complex autoimmune disease known to have an association with the HLA system. The aims of the present study were to compare the suballelic association of HLA-B27 and other HLA class I genes with microsatellite markers spanning the HLA class I region in the South Indian population of Kerala. The five microsatellites were C1_2_A (D6S2793), C1_2_5 (D6S2811), C1_4_1 (D6S2927), MIB (D6S2810), and STR-MICA. HLA typing was performed in 67 SpA patients and 77 ethnically matched healthy controls by polymerase chain reaction using sequence-specific primers, whereas fluorescence labeled microsatellites were analyzed using GeneScan analysis. There was a significant association of HLA-B27 and Cw*02 with SpA, whereas B*44 had a negative association with the disease. Only two HLA-B27 subtypes, B*2704 and B*2705, were observed in the South Indian population. We were able to successfully predict the major B27 subtype B*2705 based on the C1_2_5 microsatellite. A significant association of different alleles of all the microsatellite markers with SpA was observed. An extended six-locus haplotype, B*2705-Cw*02-STR-MICA(A4)-C1_4_1 (213 bp)-C1_2_5 (178 bp)-MIB (340 bp), was significantly associated with SpA. PMID- 16720214 TI - HLA-DRB1 alleles and cervical squamous cell carcinoma: experimental study and meta-analysis. AB - Polymorphisms in human leukocyte antigen (HLA) genes have been implicated in the risk for cervical cancer. Association of certain class II HLA alleles with cervical cancer has been documented in various ethnic populations. The implications of such an association, however, are controversial. We analyzed 126 Chinese women with cervical squamous cell carcinoma (CSCC) and 289 healthy controls to test associations of certain HLA-DRB1 alleles. We then performed meta analyses combining our own experimental data and data from nine other published studies. We found no significant differences in HLA-DRB1 allele frequencies in both CSCC and HPV-16-positive CSCC patients and control subjects. Meta-analysis provided evidence that four allele families (HLA-DRB1*04, *07, *11, and *15) and seven alleles (HLA-DRB1*0403, *0405, *0407, *0701, *1501, *1502, and *1503) were positively associated and two allele families (HLA-DRB1*09 and *13) and four alleles (HLA-DRB1*0901, *1301, *1302, and *1602) were negatively associated with CSCC in all studies or in Caucasian subgroups. In conclusion, our meta-analysis confirms the apparent association between certain HLA-DRB1 allele families and alleles and CSCC, suggesting that oncogenesis in this disease may be related to defects in immunoregulation. Larger studies may be needed, particularly in various ethnic groups, to further substantiate these associations. PMID- 16720213 TI - Genetic markers linked to rheumatoid arthritis are also strongly associated with articular manifestations in ulcerative colitis patients. AB - Ulcerative colitis is often accompanied by the development of extraintestinal, mainly articular, manifestations. Genetic differences could be underlying that clinical heterogeneity. We performed a case-control study to determine whether TNFab microsatellites or HLA-DR alleles were associated with the development of articular manifestations in patients with ulcerative colitis. With that aim, a total of 84 ulcerative colitis patients with articular manifestations and 172 without them were genotyped for TNFab microsatellites and HLA-DR. A healthy control sample (n = 595) was also included for comparative purposes. Haplotypes were inferred with the Arlequin software. The influence of HLA-DRB1*0103 and HLA B27, factors previously known to be associated with extraintestinal manifestations, was specifically addressed. We observed that TNFa6b5 minihaplotype increases the susceptibility to developing articular manifestations in ulcerative colitis patients (p = 0.003, OR = 2.39). The locus HLA-DR does not appear to be involved in these extraintestinal manifestations by itself; however, the frequency of subjects carrying TNFa6b5 in combination with DR1, DR7, or DR11 is very significantly increased in patients with articular manifestations (p = 3.9 x 10(-8)). The associations found were independent of DRB1*0103 and HLA-B27. Thus, it seems that the development of articular manifestations in ulcerative colitis patients appears to be influenced by some genetic factor(s) present in some major histocompatibility complex haplotypes. PMID- 16720215 TI - Association analysis of MYO9B gene polymorphisms with celiac disease in a Swedish/Norwegian cohort. AB - Association between myosin IXB (MYO9B) gene variants and celiac disease (CD) has been reported in a study of a Dutch cohort. Six single nucleotide polymorphisms (SNPs) within the 3' part of the MYO9B gene showed significant genetic association and formed an associated haplotype. The current study aimed to replicate these findings in a Swedish/Norwegian cohort. Genotyping of the three SNPs which tagged the associated haplotype was performed in a CD family dataset (n = 326) and in an additional set of healthy controls (n = 562). Although our material provided reasonable power to detect the previously observed association, we were unable to replicate association with these SNPs. Lack of reproducibility could be explained by no or negligible contribution of MYO9B to the genetic predisposition to CD in the Swedish/Norwegian population. Alternatively, it might be due to variable linkage disequilibria in distinct populations in the tested SNPs and a causative mutation yet to be identified or to false positive findings (type I error) in the Dutch study. PMID- 16720216 TI - The association and linkage of the HLA-A2 class I allele with autism. AB - Previous research has revealed associations between autism and immune genes located in the human leukocyte antigen (HLA). To better understand which HLA genetic loci may be associated with autism, we compared the class I HLA-A and -B alleles in autistic probands with case control subjects from Caucasian families. The frequency of HLA-A2 alleles was significantly increased in autistic subjects compared with normal allelic frequencies from the National Marrow Donors Program (NMDP) (p = 0.0043 after allelic correction). The transmission disequilibrium test for the A2 allele revealed an increased frequency of inheritance for autistic children (p = 0.033). There were no significant associations of autism with HLA-B alleles; however, the A2-B44 and A2-B51 haplotypes were two times more frequent in autistic subjects. The association and linkage of the class I HLA-A2 allele with autism suggests its involvement in the etiology of autism. Possible roles are discussed for the HLA-A2 association in the presentation of microbial antigen within the central nervous system and/or in the establishment of synaptic and neuronal circuits in the developing brain. PMID- 16720217 TI - HLA-DQ polymorphism in Turkish patients with myasthenia gravis. AB - Genetic susceptibility to myasthenia gravis (MG) is reported frequently and varies depending on the clinical presentation of the patients. HLA-DQ genotyping was performed in 132 patients using polymerase chain reaction and sequence specific oligonucleotide hybridizations in the Turkish population for the first time in this study. Antibody positivities against acetylcholine receptor and titin were 81 and 27%, respectively. Sixty-five percent of the patients had disease onset before 40 years of age (EOMG). Overall distribution of DQA1*0103 (odds ratio (OR): 0.5) and DQB1*0502 (OR: 1.9) alleles was different in patients and an ethnically matched healthy control group. Among the subgroups, DQB1*02 was significantly more frequent in EOMG (OR: 1.8), in women with MG (OR: 2.4), and in women with EOMG (OR: 2.8), whereas DQA1*0102 and DQB1*502 (OR: 2.3 for both) were increased and DQA1*0103 (OR: 0.04) was decreased in men with MG. Seropositivity was associated with both DQA1*03 (OR: 12.1) and DQB1*0302 (OR: 14.2) in the patient group. DQA1*02 (OR: 4.9) was associated with the presence of anti-titin antibodies, whereas DQA1*0101 (OR: 3.7) and *0102 (OR: 2.9) were more frequent in patients without this antibody. The presence of thymoma in MG was positively associated with DQB1*0301 (OR: 2.8), and DQB1*02 (OR: 0.3) was significantly less frequent in this group. The HLA-DQ associations in subgroups of MG suggest that the heterogeneity of the disease may be influenced by different genes or even by different alleles. DQ alleles have proved to be relatively informative polymorphisms in studying MG. PMID- 16720218 TI - Identification of two novel HLA-C alleles, Cw*070105 and Cw*1408, from East African women. AB - HLA-C was shown to be a highly polymorphic gene that can be accurately typed by sequencing methodologies. We report two novel HLA-C alleles identified during sequence-based typing of East African populations; the novel alleles were confirmed by sequencing two separate polymerase chain reaction products and by molecular cloning and sequencing multiple clones. The first new allele is identical to Cw*0701 except for a single-nucleotide synonymous substitution at codon 182 (GCA-->GCG). The new allele has been named by the WHO nomenclature committee as Cw*070105. The second new allele is identical to Cw*1403 except for a nonsynonomous change at codon 21 (CAC-->CGC), changed from histidine to arginine. The new allele has been named by the WHO nomenclature committee as Cw*1408. PMID- 16720219 TI - Direct determination of single nucleotide polymorphism haplotype of NFKBIL1 promoter polymorphism by DNA conformation analysis and its application to association study of chronic inflammatory diseases. AB - We previously revealed that one of the human leukocyte antigen-linked susceptibility genes for Takayasu's arteritis (TA) was mapped between TNFA and MICB loci and that -63T allele of NFKBIL1, which is between TNFA and MICB loci, was associated with rheumatoid arthritis (RA) in the Japanese population. We have developed a novel typing method based on reference strand-mediated conformation analysis for the upstream sequence of the NFKBIL1 gene, where -422 (T)8/(T)9, 325 C/G, -263 A/G, and -63 T/A polymorphisms were found. Upon the analysis of the patients with TA (n = 84), those with RA (n = 120), and healthy control subjects (n = 217), five common haplotypes named IKBLp*01 through IKBLp*05 were found in the Japanese population. The frequency of IKBLp*03 was significantly increased in the patient with TA (57.1% vs 35.0%, giving an odds ratio of 2.47). In addition, the frequency of IKBLp*01, but not that of other -63T-bearing alleles, was increased in the patients with RA (73.3% vs 58.1%, giving an odds ratio of 1.99), suggesting that the susceptibility to RA was conferred not by -63T alone but by combination of single nucleotide polymorphisms in the NFKBIL1 promoter. A higher promoter activity associated with IKBLp*03 and a lower activity associated with IKBLp*01 may contribute to the susceptibility to TA and RA, respectively. PMID- 16720220 TI - Real-time PCR using fluorescent resonance emission transfer probes for HLA-B typing. AB - HLA genotyping by polymerase chain reaction (PCR) has some inherent labor intensive and effort-demanding limitations. To overcome them, we have developed a real-time PCR with hybridization probes approach able to obtain a medium-low resolution HLA-B genotyping with fewer tubes and probes and with a shorter time requirement. Our strategy used 18 simultaneous reactions amplifying HLA-B alleles and an internal control. Monitorization of both amplifications in each tube is performed by the simultaneous application of two fluorescent resonance emission transfer probes: the first probe, different for each tube, is specific for the HLA-B locus and the second probe detects the control gene. A medium-low resolution (300 HLA-B allelic groups) typing is obtained for each sample by analyzing the melting curve patterns. Because some alleles may be determined without using the complete set of reactions, we present an alternative strategy: a first round of seven reactions and, according to the result, a second (or third) round of PCRs to solve the ambiguities. This method was validated in pretyped clinical samples and the results were completely concordant. Moreover, fewer ambiguous results were obtained. In summary, we present a new, faster, and more accurate method than currently used PCR techniques to type HLA-B alleles. PMID- 16720221 TI - White matter T2 hyperintensity development and clinical deterioration after status epilepticus in a patient with dentatorubral-pallidoluysian atrophy. AB - Serial T2-weighted magnetic resonance imaging in a 29-year-old woman with juvenile type dentatorubral-pallidoluysian atrophy (DRPLA) demonstrated that a cerebral white matter hyperintensity appeared within 2 months after status epilepticus and persisted for more than 20 months. The patient had rapidly progressive mental regression and became akinetic after status epilepticus. The chronological relationship between the signal changes and the clinical deterioration suggested that the epilepsy, at least in part, contributed to the progression of white matter degeneration, the hallmark of DRPLA. PMID- 16720222 TI - Intravascular lymphomatosis mimicking disseminated encephalomyelitis and encephalomyelopathy. AB - Intravascular lymphomatosis is characterized by the presence of large lymphoma cells predominantly within small vessels. This report presents two patients with diagnostically misleading neurological manifestation of this disease. Case 1, a 63-year-old man, developed a sensorimotor transverse spinal cord syndrome and encephalopathy. Lumbar puncture revealed albuminocytological dissociation. Magnetic resonance imaging (MRI) showed progression of multifocal infarct-like lesions in the brain, the thoracic cord and the medullary cone. Autoimmune inflammation was suspected, and the patient received immunosuppressive therapy with immunoglobulins, steroids and azathioprine. He died 18 months after the onset of symptoms. Case 2, a 68-year-old man, showed fluctuating aphasia, disorientation, and fever for several months. Brain MRI-scan, electroencephalography (EEG) and cerebrospinal fluid (CSF) cytology were inconclusive. Premortal biopsy of lesions in liver and right suprarenal gland showed no further characterized malignancy. He died 6 months after the first occurrence of symptoms. Autopsy of both cases revealed an intravascular lymphomatosis. Tumour cells were seen disseminated in extranodal sites including heart, lung, adrenal gland, spleen, thyroid gland and brain. An intravascular lymphomatosis should be considered when a meningoencephalitic symptomatology is unclear. A biopsy of different organs including the brain and leptomeninges should not be delayed to ensure ante mortem diagnosis and to initiate chemotherapy. PMID- 16720223 TI - Penetrating craniocerebral injury caused by a pneumatic nail gun: an unsuccessful attempt of suicide. AB - Nail guns are powerful tools commonly used in the building industry. As a result of their improper use, many accidents of bodily injury, including death, have already been reported over the last 50 years; their use in suicide attempts, however, is rare. In this paper, an unusual case of unsuccessful suicidal craniocerebral penetrating injury committed with a pneumatic nail gun by a 23 year-old man is presented. The particular findings that suggest a suicidal attempt are also discussed. PMID- 16720224 TI - Nocardia abscess during treatment of brain toxoplasmosis in a patient with aids, utility of proton MR spectroscopy and diffusion-weighted imaging in diagnosis. AB - We report the case of a 30-year-old man with known HIV-positive status who developed, 4 months prior to admission, recurrent left partial motor seizures followed by left hemiparesis. At another hospital, contrasted CT scan of the head revealed right frontal hypodense lesion with mass effect and focal contrast enhancement. A small left occipital lesion was also present. HIV-associated brain toxoplasmosis was considered and phenytoin, pyrimethamine, clindamycin and antiretrovirals were administered. Hemiparesis improved but, 3 weeks prior to admission, he developed progressive headache and bilateral visual defects. Upon admission to our center, he was found with left homonymous hemianopsia, right hemiparesis and a large hypodense left occipital lesion on a head CT scan. Proton MR spectroscopy showed lactate at 1.3ppm, amino acids at 0.9ppm, and diffusion weighted imaging (DWI) revealed hyperintensity at the lesion, suggesting a pyogenic abscess. Aspiration yielded purulent material and Nocardia asteroides grew in culture. The patient was treated with trimethoprim-sulfametoxazole and recovered with a mild visual field residual defect. PMID- 16720225 TI - Superficial siderosis of the CNS: report of three cases and review of the literature. AB - Superficial siderosis (SS) is a rare entity. A chronic subarachnoid hemorrhage (SAH) leads to the deposition of ferric pigments and ions in the surface of the central nervous system (CNS). The cause of this hemorrhage is identified in half of the cases. The disease progresses slowly and the damage is often irreversible by the time diagnosis is established. The management focuses on the identification and ablation of the bleeding source. Knowledge about this entity is based on sporadic reports. We describe three cases of superficial siderosis and briefly review the literature. PMID- 16720226 TI - Cheiro-oral-pedal syndrome due to brainstem hemorrhage. AB - Cheiro-oral-pedal syndrome is characterized by specific sensory disturbance around the corner of the mouth, in the hand and in the foot on the same side. Lesions responsible for causing this syndrome vary. We report two cases of cheiro oral-pedal syndrome due to midbrain and pontine hemorrhage, respectively. Pontine hemorrhage producing cheiro-oral-pedal syndrome has been reported in three cases, but this is the first case that midbrain hematoma exhibits this syndrome. Damage in the sensory pathway can cause cheiro-oral-pedal syndrome. Difference in the threshold may explain the specific sensory pattern in this syndrome. Cheiro-oral pedal syndrome is caused by lacunar infarction in majority of the cases. However, it should be kept in mind that hematomas can cause cheiro-oral-pedal syndrome. PMID- 16720228 TI - Overview of non-Hodgkin's lymphoma. AB - OBJECTIVES: To provide an overview of non-Hodgkin's lymphoma (NHL). DATA SOURCES: Review and research articles. CONCLUSION: NHLs are divided primarily into two main categories based on their rate of growth: aggressive and indolent. Within these two groups are various subtypes that have various clinical features. Prognostic features have been useful in determining the potential outcome of treatment of patients with NHL. IMPLICATIONS FOR NURSING PRACTICE: Nurses play an important role in the education of patients regarding the type of lymphoma they have and the steps necessary to make an accurate diagnosis, which is important in determining the appropriate treatment. PMID- 16720229 TI - Mucosa-associated lymphoid tissue (MALT) lymphoma. AB - OBJECTIVES: To review the pathogenesis of MALT lymphoma, and the associations with antigenic stimulation, presentation, and unique treatment. DATA SOURCES: Research and review articles and textbooks. CONCLUSION: MALT lymphoma is a heterogeneous but unique pathologic form of extranodal B-cell non-Hodgkin's lymphoma. The gastrointestinal tract is the most common site of disease, but involvement of multiple other organ systems has been reported. In the case of gastric MALT lymphomas, H. pylori has been highly implicated as the stimulating agent. Treatment strategies for early disease involve eradication of H. pylori when it is involved and/or radiotherapy, which alone may result in cure of this lymphoma. Chemotherapy is used in more advanced or resistant disease. IMPLICATIONS FOR NURSING PRACTICE: Nurses play a significant role in the education of patients about this unique malignancy and follow-up regarding compliance with therapy. Attention to social needs of the patients is critical. PMID- 16720230 TI - AIDS-related lymphoma. AB - OBJECTIVE: To review the epidemiology, pathology, clinical features, prognostic factors, and treatment approaches of patients with AIDS-related lymphoma. DATA SOURCES: Research studies and review articles. CONCLUSION: Aggressive B-cell lymphoma has become one of the more common of the initial AIDS-defining illnesses in the United States. Median survival of affected patients has improved considerably with the use of highly active anti-retroviral therapy directed against human immunodeficiency virus, along with multi-agent chemotherapy, and outcome of such patients now approaches that of human immunodeficiency virus negative patients with aggressive lymphoma. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses must be knowledgeable of AIDS-related lymphoma to provide supportive care to this patient population. PMID- 16720231 TI - Cutaneous T-cell lymphoma. AB - OBJECTIVE: To review recent information on the epidemiology, etiology, clinical presentation, staging, treatment approaches, and nursing management of CTCL. DATA SOURCES: Research and review articles and textbooks. CONCLUSION: Identifying the etiology and finding a cure for cutaneous T-cell lymphoma is promising as our knowledge and understanding of the T-cell biology evolves, creating more options for innovative therapies or treatment modalities. IMPLICATIONS FOR NURSING PRACTICE: It is essential for the oncology nurse to understand the disease process, treatment modalities, and side effect profiles to provide astute nursing assessment, establishment of treatment goals, management of cutaneous symptoms, treatment of side effects, and pain control while preventing infection and providing psychological/social support throughout the course of the disease. PMID- 16720232 TI - Treatment approaches and nursing considerations for non-Hodgkin's lymphoma. AB - OBJECTIVE: To discuss some common non-Hodgkin's lymphoma subtypes, and current systemic approaches including chemotherapy, monoclonal antibodies, radioimmunotherapy, and vaccines. DATA SOURCES: Research and review articles, textbooks, and guidelines CONCLUSION: Non-Hodgkin's lymphoma is one of the most common types of malignancies for both men and women. It manifests itself in the lymph nodes, bone marrow, and extranodal sites, with or without B symptoms (fevers, sweats, weight loss) and it continues to be on the rise. Recent advances in treatment options have improved the potential for cure within its various subtypes, although much more work remains to be done. IMPLICATIONS FOR NURSING PRACTICE: Management of side effects remains a major challenge to nurses when caring for patients undergoing treatment. PMID- 16720234 TI - Survivorship: finding a new balance. AB - OBJECTIVES: To offer personal and professional perspectives on survivorship. DATA SOURCES: Personal experience, publications, government resources. CONCLUSION: Challenges experienced by the cancer patient are influenced by numerous factors, including age, type and extent of cancer, treatment schedule and effects, financial well-being, access to support and follow-up, information needs, and family reactions. Challenges are eased through a kind word and guidance from those whom patients lean on the most and respect, the nurse. IMPLICATIONS FOR NURSING PRACTICE: Nurses play an important role in preparing the patient and their families to have realistic expectations and balance the demands along the journey of survivorship. PMID- 16720233 TI - Hematopoietic cell transplantation in non-Hodgkin's lymphoma. AB - OBJECTIVE: To explore the role and outcomes using hematopoietic cell transplantation (HCT) as a treatment option with aggressive, follicular, mantle, T-cell, and HIV related non-Hodgkin's lymphoma (NHL). DATA SOURCES: Research and review articles and textbooks. CONCLUSION: High-dose chemotherapy and/or radiation therapy followed by HCT has been used to overcome resistance to standard-dose therapy and has been explored over the past 40 years and has shown long-term survival of approximately 10% to 50% in patients with relapsed or refractory lymphoma. IMPLICATIONS FOR NURSING PRACTICE: Nursing plays a significant role in the assessment and management of patients throughout the course of HCT. PMID- 16720235 TI - Discussion of targeting proteins in vivo: in vitro guidelines. PMID- 16720236 TI - Development of 111In-labeled tumor-associated antigen peptides for monitoring dendritic-cell-based vaccination. AB - Dendritic cells (DC) are professional antigen-presenting cells capable of inducing potent immune responses. In our ongoing clinical trials, human leukocyte antigen (HLA)-A2.1+ melanoma patients are vaccinated with mature DC, presenting tumor-derived peptides in major histocompatibility complexes (MHC) to naive T cells. Previously, we have shown that both intradermally and intranodally injected (111)In-labeled mature DC migrate to draining lymph nodes. However, little is known about the fate of the MHC-peptide complex after injection of these peptide-loaded DC. The aim of the present study was to develop radiolabeled, tumor-derived peptides to monitor their binding to MHC Class I. METHODS: The HLA-A2.1 binding peptide gp100:154-162mod (gp100:154m) was conjugated with diethylenetriamine pentaacetic acid (DTPA) either at the N terminus (alpha-DTPA-gp100:154m) or at the epsilon amino group of the Lys(154) residue (epsilon-DTPA-gp100:154m) and labeled with (111)In. RESULTS: The maximum specific activity for both peptides was 13 GBq/micromol. The IC50 of the alpha [(111)In]DTPA-gp100:154m peptide was >75 microM. The IC50 of the (111)In-labeled epsilon-DTPA-gp100:154m was 3 microM, similar to the unconjugated peptide. MHC binding studies showed specific binding of the epsilon-[(111)In]DTPA-gp100:154m peptide to the JY cells at 4 degrees C. Interestingly, no specific binding was observed for the alpha-[(111)In]DTPA-gp100:154m peptide. In contrast to the alpha [(111)In]DTPA-gp100:154m peptide, the epsilon-[(111)In]DTPA-gp100:154m peptide was recognized by cytotoxic T cells. CONCLUSION: When DTPA was conjugated to the epsilon NH2 group of the Lys(154) residue, MHC binding of the peptide was preserved and could still be recognized by cytotoxic T cells. These studies allow the noninvasive determination of the behavior of MHC-peptide complexes on DC in vivo. PMID- 16720237 TI - In vitro and in vivo evaluation of novel ligands for radioimmunotherapy. AB - Novel ligands cis-2,6-bis[N,N-bis(carboxymethyl)aminomethyl]-1-piperidineacetic acid (PIP-DTPA), cis-[(1R,11S)-6,9,15-Tris-carboxymethyl-3,6,9,15 tetraazabicyclo[9.3.1]pentadec-3-yl]-acetic acid (PIP-DOTA), cis-{2,7-bis-[bis carboxymethyl-amino)-methyl]-azepan-1-yl}-acetic acid (AZEP-DTPA), [2-(4,7-bis carboxymethyl-[1,4,7]triazacyclononan-1-yl-ethyl]-2-carbonylmethyl-amino] tetraacetic acid (NETA) and [{4-carboxymethyl-7-[2-(carboxymethylamino)-ethyl] perhydro-1,4,7-triazonin-1-yl}-acetic acid (NPTA) are investigated as potential chelators of 177Lu, 90Y, 212Pb and 213Bi for radioimmunotherapy (RIT). The new ligands are radiolabeled with 177Lu, 86/88/90Y, 203Pb and 205/6Bi, and in vitro stability and in vivo stability of the radiolabeled complexes are assessed in human serum and athymic mice, respectively. In vitro studies indicate that all radiolabeled complexes with the exception of 90Y-AZEP-DTPA are stable in serum for 5-11 days. All new ligands examined herein are found to tightly hold 177Lu in vivo. Piperidine-backboned DTPA (PIP-DTPA) complexes radiolabeled with all radioisotopes examined display excellent in vivo stability, that is, excretion without dissociation. The azepane-backboned DTPA derivative, AZEP-DTPA, appears ineffective in binding all but 177Lu in vivo. NETA and NPTA radiolabeled with 86Y or 177Lu exhibit rapid blood clearance and low organ uptakes. Significant accretion in the kidney, femur and/or liver is observed with 203Pb-labeled AZEP DTPA, PIP-DOTA and NPTA. Both 203Pb-PIP-DOTA and 205/6Bi-PIP-DOTA result in moderate to high renal accumulation of radioactivity. NETA exhibits improved renal accumulation with respect to PIP-DOTA for 205/6Bi but also shows significant liver uptake. Of all ligands studied, only PIP-DTPA appears to effectively bind 203Pb and 205/6Bi in vivo. PIP-DTPA, PIP-DOTA, NETA and NPTA all show strong evidence of rapid blood clearance and low organ uptake for 177Lu and 90Y. Serum stability and in vivo biodistribution results suggest PIP-DTPA as a potential chelating agent with broad applicability for use in 177Lu, 90Y, 212Pb and 213Bi RIT. PMID- 16720238 TI - Preparation and in vivo evaluation of a novel stabilized linker for 211At labeling of protein. AB - Significant improvement of in vivo stability of 211At-labeled radioimmunoconjugates achieved upon employment of a recently reported new linker, succinimidyl N-2-(4-[211At]astatophenethyl)succinamate (SAPS), prompted additional studies of its chemistry. The 211At radiolabeling of succinimidyl N-2 (4-tributylstannylphenethyl)succinamate (1) was noted to decline after storage at -15 degrees C for greater than 6 months. Compound 1 was found to degrade via a ring closure reaction with the formation of N-2-(4 tributylstannylphenethyl)succinimide (3), and a modified procedure for the preparation of 1 was developed. The N-methyl structural analog of 1, succinimidyl N-2-(4-tributylstannylphenethyl)-N-methyl succinamate (SPEMS), was synthesized to investigate the possibility of improving the stability of reagent-protein linkage chemistry. Radiolabeling of SPEMS with 211At generates succinimidyl N-2-(4 [211At]astatophenethyl)-N-methyl succinamate (Methyl-SAPS), with yields being consistent for greater than 1 year. Radiolabelings of 1 and SPEMS with 125I generated succinimidyl N-2-(4-[125I]iodophenethyl)succinamate (SIPS) and succinimidyl N-2-(4-[125I]iodophenethyl)-N-methyl succinamate (Methyl-SIPS), respectively, and showed no decline in yields. Methyl-SAPS, SAPS, Methyl-SIPS and SIPS were conjugated to Herceptin for a comparative assessment in LS-174T xenograft-bearing mice. The conjugates of Herceptin with Methyl-SAPS or Methyl SIPS demonstrated immunoreactivity equivalent to if not superior to the SAPS and SIPS paired analogs. The in vivo studies also revealed that the N-methyl modification resulted in a superior statinated product. PMID- 16720239 TI - In vitro and in vivo evaluation of 177Lu- and 90Y-labeled E. coli heat-stable enterotoxin for specific targeting of uroguanylin receptors on human colon cancers. AB - The human E. coli heat-stable enterotoxin (ST(h), amino acid sequence N1SSNYCCELCCNPACTGCY19) binds specifically to the guanylate cyclase C (GC-C) receptor, which is present in high density on the apical surface of normal intestinal epithelial cells as well as on the surface of human colon cancer cells. Analogs of ST(h) are currently being used as vectors targeting human colon cancers. Previous studies in our laboratory have focused on development of 111Indium-labeled ST(h) analogs for in vivo imaging applications. Here, we extend the scope of this work to include targeting of the therapeutic radionuclides 90Y and 177Lu. The peptide DOTA-F19-ST(h)(1-19) was synthesized using conventional Fmoc-based solid-phase techniques and refolded in dilute aqueous solution. The peptide was purified by RP-HPLC and characterized by MALDI-TOF MS and in vitro receptor binding assay. The DOTA-conjugate was metallated with nonradioactive Lu(III)Cl3 and Y(III)Cl3, and IC50 values of 2.6+/-0.1 and 4.2+/-0.9 nM were determined for the Lu- and Y-labeled peptides, respectively. 177Lu(III)Cl3 and 90Y(III)Cl3 labeling yielded tracer preparations that were inseparable by C18 RP HPLC, indicating that putative differences between Lu-, Y- and In coordination spheres are not observed in the context of labeled ST(h) peptides. In vivo biodistribution studies of the 177Lu-labeled peptide in severe combined immunodeficient (SCID) mice bearing T-84 human cancer tumor xenografts showed rapid clearance from the bloodstream, with >90 %ID in the urine at 1 h pi. Localization of the tracer within tumor xenografts was 1.86+/-0.91 %ID/g at 1 h pi, a value higher than for all other tissues with the exception of kidney (2.74+/-0.24 %ID/g). At 24 h pi, >98 %ID was excreted into the urine, and 0.35+/ 0.23 %ID/g remained in tumor, again higher than in all other tissues except kidney (0.91+/-0.46 %ID/g). Biodistribution results at 24 h pi for the 90Y labeled peptide mirrored those for the 177Lu analog, in agreement with the identical behavior of the labeled analogs by C18 RP-HPLC. These results demonstrate the ability of 177Lu- and 90Y-labeled ST(h) molecules to specifically target GC-C receptors expressed on T-84 human colon cancer cells. PMID- 16720240 TI - Development of a 111In-labeled peptide derivative targeting a chemokine receptor, CXCR4, for imaging tumors. AB - The chemokine receptor CXCR4 is highly expressed in tumor cells and plays an important role in tumor metastasis. The aim of this study was to develop a radiopharmaceutical for the imaging of CXCR4-expressing tumors in vivo. Based on structure-activity relationships, we designed a 14-residue peptidic CXCR4 inhibitor, Ac-TZ14011, as a precursor for radiolabeled peptides. For 111In labeling, diethylenetriaminepentaacetic acid (DTPA) was attached to the side chain of d-Lys(8) which is distant from the residues indispensable for the antagonistic activity. In-DTPA-Ac-TZ14011 inhibited the binding of a natural ligand, stromal cell-derived factor-1alpha, to CXCR4 in a concentration-dependent manner with an IC50 of 7.9 nM (Ac-TZ14011: 1.2 nM). In biodistribution experiments, more 111In-DTPA-Ac-TZ14011 accumulated in the CXCR4-expressing tumor than in blood or muscle. Furthermore, the tumor-to-blood and tumor-to-muscle ratios were significantly reduced by coinjection of Ac-TZ14011, indicating a CXCR4-mediated accumulation in tumor. These findings suggested that 111In-DTPA-Ac TZ14011 would be a potential agent for the imaging of CXCR4 expression in metastatic tumors in vivo. PMID- 16720241 TI - Double-stabilized neurotensin analogues as potential radiopharmaceuticals for NTR positive tumors. AB - INTRODUCTION: Overexpression of neurotensin (NT) receptors in exocrine pancreatic cancer and other neuroendocrine cancers make them interesting targets for tumor imaging and therapy. Modifications at the cleavage bonds 8-9 and 11-12 led to the synthesis of NT-XII, NT-XIII and NT-XVIII, three new stabilized analogues. (NalphaHis)Ac was coupled to the N-terminus for labeling with [(99m)Tc] tricarbonyl. METHODS: Stability was tested in vitro in human plasma and HT-29 cells. Binding to NT1 receptors and internalization/efflux were analyzed in intact HT-29 cells. Biodistribution studies were performed in nude mice bearing HT-29 xenografts. RESULTS: All analogues were very stable in human plasma, with half-lives of 20-21 days. Degradation in HT-29 cells was more rapid (t(1/2) of 6.5, 5 and 2.5 h for NT-XII, NT-XIII and NT-XVIII, respectively). They also showed high affinity and specificity for NT1 receptors. Bound activity was rapidly internalized at 37 degrees C. The pattern of externalization was different. NT-XII was released more slowly than NT-XIII and NT-XVIII (half of the activity still inside the cells after 24 h). Bigger differences were found in the biodistribution studies. NT-XII showed the highest tumor uptake as well as the best tumor to nontumor ratios. CONCLUSION: The modifications introduced in NT(8 13) increased plasma stability, maintaining unaffected the in vitro binding properties. The best biodistribution corresponded to NT-XII, which shows to be a good candidate for NT1 receptors overexpressing tumors. First clinical trials are ongoing. PMID- 16720242 TI - Radionuclide imaging of small-cell lung cancer (SCLC) using 99mTc-labeled neurotensin peptide 8-13. AB - OBJECTIVES: To prepare 99m technetium (99mTc)-labeled neurotensin (NT) peptide and to evaluate the feasibility of imaging oncogene NT receptors overexpressed in human small-cell lung cancer (SCLC) cells. METHODS: The NT analogue (Nalpha His)Ac-NT(8-13) was synthesized such that histidine was attached at the N terminus. The analogue was labeled with [99mTc(H2O)3(CO)3] at pH 7. 99mTc-(Nalpha His)Ac-NT(8-13) in vitro stability was determined by challenging it with 100 times the molar excess of DTPA, human serum albumin (HSA) and cysteine. The affinity, 99mTc-(Nalpha-His)Ac-NT(8-13) binding to SCLC cell line NCI-H446, was studied in vitro. Biodistribution and imaging with 99mTc-(Nalpha-His)Ac-NT(8-13) were performed at 4 and 12 h postinjection, and tissue distribution and imaging after receptor blocking were carried out at 4 h in nude mice bearing human SCLC tumor. Blood clearance was determined in normal mice. RESULTS: The affinity constant (Kd) of 99mTc-(Nalpha-His)Ac-NT(8-13) to SCLC cells was 0.56 nmol/L. When challenged with 100 times the molar excess of DTPA, HSA or cysteine, more than 97+/-1.8% radioactivity remained as 99mTc-(Nalpha-His)Ac-NT(8-13). Tumor-to muscle ratio was 3.35+/-1.01 at 4 h and 4.20+/-1.35 at 12 h postinjection. The excretory route of 99mTc-(Nalpha-His)Ac-NT(8-13) was chiefly through the renal pathway. In the receptor-blocking group treated with unlabeled (Nalpha-His)Ac NT(8-13), tumor-to-muscle ratio at 4 h was 1.25+/-0.55. CONCLUSION: The results suggest that 99mTc-(Nalpha-His)Ac-NT(8-13) specifically binds to the SCLC cells and made 99mTc-(Nalpha-His)Ac-NT(8-13) a desirable compound for further studies in planar or SPECT imaging of oncogene receptors overexpressed in SCLC cells. PMID- 16720243 TI - Rhemium-186-monoaminemonoamidedithiol-conjugated bisphosphonate derivatives for bone pain palliation. AB - To develop a radiopharmaceutical for the palliation of painful bone metastases based on the concept of bifunctional radiopharmaceuticals, we synthesized a bisphosphonate derivative labeled with rhenium-186 (186Re) that contains a hydroxyl group at the central carbon of its bisphosphonate structure, we attached a stable 186Re-MAMA chelate to the amino group of a 4-amino butylidene bisphosphonate derivative [N-[2-[[4-[(4-hydroxy-4,4-diphosphonobutyl)amino]-4 oxobutyl]-2-thioethylamino]acetyl]-2-aminoethanethiolate] oxorhenium (V) (186Re MAMA-HBP) and we investigated the effect of a hydroxyl group at the central carbon of its bisphosphonate structure on affinity for hydroxyapatite and on biodistribution by conducting a comparative study with [N-[2-[[3-(3,3 diphosphonopropylcarbamoyl)propyl]-2-thioethylamino]acetyl]-2 aminoethanethiolate] oxorhenium (V) (186Re-MAMA-BP). The precursor of 186Re-MAMA HBP, trityl (Tr)-MAMA-HBP, was obtained by coupling a Tr-MAMA derivative to 4 amino-1-hydroxybutylidene-1,1-bisphosphonate. 186Re-MAMA-HBP was prepared by a reaction with 186ReO(4-) and SnCl2 in citrate buffer after the deprotection of the Tr groups of Tr-MAMA-HBP. After reversed-phase high-performance liquid chromatography, 186Re-MAMA-HBP had a radiochemical purity of over 95%. Compared with 186Re-MAMA-BP, 186Re-MAMA-HBP showed a greater affinity for hydroxyapatite beads in vitro and accumulated a significantly higher level in the femur in vivo. Thus, the introduction of a hydroxyl group into 186Re complex-conjugated bisphosphonates would be effective in enhancing accumulation in bones. These findings provide useful information on the design of bone-seeking therapeutic radiopharmaceuticals. PMID- 16720244 TI - [11C]palmitate kinetics across the splanchnic bed in arterial, portal and hepatic venous plasma during fasting and euglycemic hyperinsulinemia. AB - PURPOSE: The liver is fundamental in regulating lipid metabolism, and it supplies fatty acids (FA) to the rest of the body in the form of triglycerides (TG); the time-related relevance of this process is incompletely defined. The aim of the study was to investigate the appearance of labeled TG in the hepatic vascular bed after [11C]palmitate injection during fasting and insulin stimulation. METHODS: Plasma [11C]palmitate kinetics in arterial, portal and hepatic venous lipid fractions was studied in eight anesthetized pigs during fasting or euglycemic hyperinsulinemia. Plasma analyses were conducted at 10 and 40 min after tracer injection. Corresponding liver positron emission tomography (PET) images were acquired for the semiquantitative determination of hepatic FA uptake. RESULTS: At 10 min, plasma levels of unchanged [11C]palmitate were lower in hyperinsulinemic than in fasting experiments in the artery and in the portal vein (P< or=.03), suggesting faster clearance. Levels of unmetabolized [11C]palmitate did not differ between portal and arterial plasma. In the fasting state, a tendency to a positive arterial and portal vs. hepatic venous gradient was observed, indicative of net hepatic [11C]palmitate extraction. Labeled TG were already detectable at 10 min (fasting vs. hyperinsulinemia, ns) and were higher in fasting than in hyperinsulinemic animals at 40 min (92+/-1% and 82+/-6% of arterial plasma radioactivity). Higher proportions of labeled TG were recovered in portal vein plasma, suggesting release by the gut. The portal and the arterial-portal vs. hepatic venous TG gradient tended to be positive. Accordingly, hepatic FA uptake was higher, but declined more rapidly during fasting than during hyperinsulinemia. CONCLUSION: The study indicates that the redistribution of [11C]palmitate between different lipid pools occurs within the short time interval of most PET experiments and is strongly influenced by insulin. Labeled TG constitute an additional [11C]palmitate source in the modeling of PET data. PMID- 16720245 TI - Asialoglycoprotein-receptor-targeted hepatocyte imaging using 99mTc galactosylated chitosan. AB - This study investigated the usefulness of 99mTc hydrazinonicotinamide galactosylated chitosan (HGC) in hepatocyte imaging. HGC was obtained by coupling the galactose moiety of both lactobionic acid and succinimidyl 6 hydrazinonicotinate hydrochloride (succinimidyl HYNIC). The coupled product was then radiolabeled with 99mTc using stannous chloride and tricine as reducing agent and coligand, respectively. Labeling efficiency was >90% both in room temperature and in serum up to 24 h after injection. The hepatic uptake properties of 99mTc HGC were studied in Balb/C mice. 99mTc HGC and 99mTc hydrazinonicotinamide chitosan (HC) were intravenously injected into mice, with receptor binding identified by coinjection with 9 and 14 mg of free galactose. Images were acquired with a gamma-camera. After injection via the tail vein of the mice, 99mTc HGC showed high selectivity for the liver, while 99mTc HC without a galactose group showed low liver uptake. In addition, the hepatic uptake of 99mTc HGC was blocked by coinjection of free galactose. Tissue distribution was determined at three different times (10, 60 and 120 min). The liver accumulated 13.16+/-2.72%, 16.11+/-5.70% and 16.55+/-2.28% of the injected dose per gram at 10, 60 and 120 min after injection, respectively. 99mTc HGC showed specific and rapid targeting of hepatocytes. It is a promising receptor-specific radiopharmaceutical with potential applications in liver imaging for the evaluation of hepatocytic function. PMID- 16720246 TI - Acute treatment with pentobarbital alters the kinetics of in vivo receptor binding in the mouse brain. AB - The effect of pentobarbital, a sedative-hypnotic barbiturate, on the in vivo binding of benzodiazepine receptors in the mouse brain was investigated. Dose related changes in the apparent binding of [3H]Ro15-1788 ([3H]flumazenil) in the cerebral cortex, cerebellum and pons-medulla were observed by pretreatment with pentobarbital. For quantification of the kinetic properties of the in vivo binding of [3H]Ro15-1788, time courses of radioactivity following its injection were examined, and kinetic analysis was performed using the compartment model. The time courses of radioactivity following injection of [3H]Ro15-1788 with 3 mg/kg Ro15-1788 were used as input function. In all regions studied, rate constants between input compartment and specific binding compartment were significantly decreased by pentobarbital. However, no significant alterations in the binding potential (BP=K3/K4) of benzodiazepine receptors by pentobarbital were observed in any of the regions. A saturation experiment indicated that the decrease in the input rate constant (K3), which includes both the association rate constant (k(on)) and the number of binding sites available (B(max)), was mainly due to decrease in k(on). These results suggest that apparent increases in binding at 20 min after tracer injection were due to the decrease in the association and dissociation rates of binding in vivo. PMID- 16720247 TI - Evaluation of (+)-p-[11C]methylvesamicol for mapping sigma1 receptors: a comparison with [11C]SA4503. AB - Vesamicol is a leading compound for positron emission tomography (PET) and single photon emission computed tomography (SPECT) tracers for mapping the vesicular acetylcholine transporter (VAChT). Recently, we found that (+)-p-methylvesamicol ((+)-PMV) has low affinity for VAChT (K(i)=199 nM), but has moderate to high affinity for sigma receptors: K(i)=3.0 nM for sigma1 and K(i)=40.7 nM for sigma2, and that sigma1-selective SA4503 (K(i)=4.4 nM for sigma1 and K(i)=242 nM for sigma2) has moderate affinity for VAChT (K(i)=50.2 nM). In the present study, we examined the potential of (+)-[11C]PMV as a PET radioligand for mapping sigma1 receptors as compared with [11C]SA4503. In rat brain, similar regional distribution patterns of (+)-[11C]PMV and [11C]SA4503 were shown by tissue dissection and by ex vivo autoradiography. Blocking experiments using (+/-)-PMV, (-)-vesamicol, SA4503, haloperidol and (+/-)-pentazocine showed that the two tracers specifically bound to sigma1 receptors, and that [11C]SA4503 exhibited greater specific binding than (+)-[11C]PMV. No sign of VAChT-specific binding by [11C]SA4503 was observed in the striatum, which is rich in VAChT sites. In conclusion, (+)-[11C]PMV specifically bound to sigma1 receptors in the brain, but to a lesser extent than [11C]SA4503, suggesting that (+)-[11C]PMV is a less preferable PET ligand than [11C]SA4503. On the other hand, the moderate affinity of [11C]SA4503 for VAChT is negligible in vivo. PMID- 16720248 TI - Chemical fate of the nicotinic acetylcholinergic radiotracer [123I]5-IA-85380 in baboon brain and plasma. AB - The fate of the nicotinic acetylcholinergic receptor radiotracer [123I]5-IA-85380 ([123I]5-IA) was studied in baboon by analyzing the chemical composition of brain tissue and plasma after intravenous administration of the tracer. Acetonitrile denaturation and high-performance liquid chromatography (HPLC) analysis showed predominantly unchanged (91-98%) parent tracer in all brain tissues examined, compared to significant metabolism (23% parent) in the plasma at 90 min postinjection, and control tissue recovery of 95-98%. [123I]5-IA was distributed to the thalamus with a standardized uptake value of 9.2 (0.04% dose/g) or a concentration 5.8 times higher than that of the cerebellum. The HPLC behavior of a synthesized sample of one hypothesized metabolite, 5-iodo-3-pyridinol (5-IP), was consistent with plasma radiometabolite fraction. Since only parent radiotracer compound was found in brain tissue, these results add confidence that information derived from single photon emission computed tomography images of 123I activity in the brain after [123I]5-IA administration can be interpreted as distribution of an intact radiotracer. PMID- 16720249 TI - The serotonin transporter in rhesus monkey brain: comparison of DASB and citalopram binding sites. AB - We have characterized the interaction of the serotonin transporter ligand [3H] N,N-dimethyl-2-(2-amino-4-cyanophenylthio)-benzylamine (DASB) with rhesus monkey brain in vitro using tissue homogenate binding and autoradiographic mapping. [3H] DASB, a tritiated version of the widely used [11C] positron emission tomography tracer, was found to selectively bind to a single population of sites with high affinity (K(d)=0.20+/-0.04 nM). The serotonin transporter density (B(max)) obtained for rhesus frontal cortex was found to be 66+/-8 fmol/mg protein using [3H]-DASB, similar to the B(max) value obtained using the reference radioligand [3H]-citalopram, a well-characterized and highly selective serotonin reuptake inhibitor (83+/-22 fmol/mg protein). Specific binding sites of both [3H]-DASB and [3H]-citalopram were similarly and nonuniformly distributed throughout the rhesus central nervous system, in a pattern consistent with serotonin transporter localization reported for human brain. Regional serotonin transporter densities, estimated from optical densities of the autoradiographic images, were well correlated between the two radioligands. Finally, DASB and fluoxetine showed dose dependent full inhibition of [3H]-citalopram binding in a competition autoradiographic study, with K(i) values in close agreement with those obtained from rhesus brain homogenates. This side-by-side comparison of [3H]-DASB and [3H] citalopram binding sites in rhesus tissue homogenates and in adjacent rhesus brain slices provides additional support for the use of [11C]-DASB to assess the availability and distribution of serotonin transporters in nonhuman primates. PMID- 16720250 TI - Synthesis and in vivo evaluation of [O-methyl-11C](2R,4R)-4-hydroxy-2-[2-[2-[2-(3 methoxy)phenyl]ethyl]phenoxy]ethyl-1-methylpyrrolidine as a 5-HT2A receptor PET ligand. AB - The serotonin2A (5-HT2A) receptor is implicated in the pathophysiology of schizophrenia and mood disorders, and in vivo studies of this receptor would be of value in studying the pathophysiology of these disorders and in measuring the relationship of clinical response to receptor occupancy for 5-HT2A antagonists such as atypical antipsychotics. Therefore, (2R,4R)-4-hydroxy-2-[2-[2-[2-(3 methoxy)-phenyl]ethyl]phenoxy]ethyl-1-methylpyrrolidine (MPM) (13), a selective and high-affinity (K(i)=0.79 nM) 5HT2A antagonist, has been radiolabeled with carbon-11 by O-methylation of the corresponding desmethyl analogue (2R,4R)-4 hydroxy-2-[2-[2-[2-(3-hydroxy)phenyl]ethyl]phenoxy]ethyl-1-methylpyrrolidine (12) with [11C]methyltriflate in order to determine the suitability of [11C]MPM to quantify 5-HT2A in living brain using PET. Desmethyl-MPM 12 and standard MPM were prepared, starting from 3-hydroxymethylphenol (2), in excellent yield. The yield obtained for radiolabeling was 40+/-5% (EOB), and the total synthesis time was 30 min at EOS. PET studies with [11C]MPM in baboon showed a distribution in the brain consistent with the known distribution of 5-HT2A receptors. The time activity curves for the high-binding regions peaked at approximately 45 min after injection. Blocking studies with M100907 demonstrated not only 38-57% blocking of tracer binding in brain regions known to have 5-HT2A receptors but also 38% blocking in cerebellum, which has a low 5-HT2A receptor concentration. Although [11C]MPM exhibits appropriate kinetics in baboon for imaging 5-HT2A receptors, its specific binding in cerebellum and higher proportion of nonspecific binding limit its usefulness for the in vivo quantification of 5-HT2A receptors with PET. PMID- 16720251 TI - 99mTc-pertechnetate uptake in hepatoma cells due to tissue-specific human sodium iodide symporter gene expression. AB - The sodium iodide symporter (NIS) gene could be used as an ideal reporter gene as well as a promising therapeutic gene. 99mTc-pertechnetate has proven to be more advantageous than 131I-iodide with respect to image quality, procedure and radiation dose in examination of thyroid uptake and scintigraphy. Herein, we investigated the feasibility of monitoring human sodium iodide symporter (hNIS) gene expression with 99mTc-pertechnetate in hepatoma cells (MH3924A) following tissue-specific expression. METHODS: MH3924A cells were stably transfected with the recombinant retroviral vector, in which hNIS cDNA was driven by murine albumin enhancer/promoter (mAlb) and coupled to hygromycin resistance gene using an internal ribosomal entry site. Functional NIS expression in hepatoma cells was confirmed by an 125I(-) uptake assay. The dynamic uptake and efflux of 99mTc pertechnetate was determined both in vitro and in vivo. RESULTS: The 99mTc pertechnetate was up to 254-fold higher in stably transfected MH3924A cells than in wild-type cells. However, the in vitro efflux of 99mTc-pertechnetate out of recombinant cells was rapid with a half-life of less than 2 min. Further, the in vivo studies yielded clear images and quantitative data of mAlbhNIS-infected tumor xenografts using 99mTc-pertechnetate and gamma camera. CONCLUSION: The current study demonstrates enhanced 99mTc-pertechnetate uptake in hepatoma cells in vitro and in vivo following tissue-specific gene transfer using a recombinant retrovirus with the albumin enhancer/promoter and the hNIS gene. It is feasible to monitor hNIS gene expression noninvasively and quantitatively using conventional gamma camera and 99mTc-pertechnetate. PMID- 16720252 TI - Effect of iron deficiency anemia on the biodistribution of 99mTc radiopharmaceuticals. AB - The distribution of colloids and labeled cells in organs is influenced by their intrinsic properties and by the state of the investigated subject. Iron deficiency remains an unsolved nutritional problem all over the world; one of its severe consequences is anemia. Because iron metabolism principally takes place in the liver, spleen, bone marrow, skeletal muscle and blood, we studied the effect of iron deficiency anemia on the biodistribution of 99mTc phytate, 99mTc gelatin colloid and 99mTc RBC (red blood cells labeled with 99mTc). Our results show that iron deficiency anemia modifies the pattern of biodistribution of the two colloids assayed. However, this behavior is different for both of them. This work contributes to studies that kinetically and statistically establish that iron deficiency anemia induces a significant inversion in the spleen-liver activity relationship when centellographic studies are performed with colloids such as 99mTc phytate. PMID- 16720253 TI - 175Yb-labeled hydroxyapatite: a potential agent for use in radiation synovectomy of small joints. AB - The preparation of 175Yb-labeled hydroxyapatite (HA) particle is described for possible use as an agent for radiation synovectomy (RS) of small-sized joints. 175Yb was produced by thermal neutron irradiation of enriched (98.6% in 174Yb) ytterbium target at a flux of approximately 3 x 10(13) n/cm(2)/s for 7 days. Specific activity of 5.5-6.0 GBq/mg and a very high radionuclidic purity to the extent of approximately 100% were obtained. In the work reported herein, HA could be labeled with 175Yb in very high radiochemical purity (>99%) using 10 mg of HA particle at pH approximately 7. The radiolabeled particulates showed excellent in vitro stability at room temperature. Serial scintigraphic images of normal as well as arthritis-bearing Wistar rats following intra-articular injection of 175Yb-HA particles in the knee joint showed complete retention of activity within the synovial cavity with no measurable activity leaching out from the joint till 144 h post-injection. PMID- 16720254 TI - Evolutionary function of dreams: A test of the threat simulation theory in recurrent dreams. AB - proposed an intriguing and detailed evolutionary theory of dreams which stipulates that the biological function of dreaming is to simulate threatening events and to rehearse threat avoidance behaviors. The goal of the present study was to test this theory using a sample of 212 recurrent dreams that was scored using a slightly expanded version of the DreamThreat rating scale. Six of the eight hypotheses tested were supported. Among the positive findings, 66% of the recurrent dream reports contained one or more threats, the threats tended to be dangerous and aimed at the dreamer, and when facing a threat, the dreamer tended to take defensive or evasive actions that were possible and reasonable. However, less than 15% of the recurrent dreams depicted realistic and probable situations critical for one's physical survival or reproductive success and the dreamer rarely succeeded in fleeing the threat despite important and appropriate efforts. The findings thus provide mixed support for the threat simulation theory. PMID- 16720256 TI - The changing face of case managers. PMID- 16720257 TI - Linking the power of information technology and case management interventions: an assessment of recent medical-management trends. AB - Imagine a case manager screening a large employer group of 20,000 participants and within less than a couple of minutes: Your first reaction may be one of disbelief. You might think, "Yes, maybe in 2020." However, the reality is that this type of information exchange in the context of case management (CM) is happening today, and it is revolutionizing the medical-management system as we once knew it. This article highlights some of the key changes that have recently occurred in the medical-management field and that are creating a paradigm shift in how CM services will be rendered in the future. PMID- 16720258 TI - Essential activities and knowledge domains of case management: new insights from the CCMC role and functions study. AB - The Commission for Case Manager Certification (CCMC) defines case management (CM) as "a collaborative process that assesses, plans, implements, coordinates, monitors, and evaluates the options and services required to meet an individual's health needs. [Case management] uses communication and available resources to promote quality, cost-effective outcomes." The practice of CM spans the entire health-care spectrum, including pre-acute, acute, and post-acute settings, and the involvement of varied care providers, such as nurses, social workers, rehabilitation counselors, physicians, and other allied health professionals. So what does it mean to practice as a case manager? What roles and job functions are performed and what knowledge is required of a professional in the field for effective practice? These highly relevant questions reflect the thinking of the CCMC commissioners when the latest Case Manager Role and Functions study was undertaken. The primary purpose of this research, which is conducted every 5 years by the CCMC, is to capture the current state of CM practice. This type of in-depth research is required to support an evidence-based certification examination such as the one offered by CCMC-the certified case manager (CCM) credential. Moreover, as the first and largest nationally accredited organization to certify US case managers, the CCMC recognizes its responsibility to undertake and promote scientifically conducted research in the field of CM. PMID- 16720259 TI - The lessons of case management. AB - The health-care system works, for the most part, and patients who need help from case management are the exception rather than the rule. This has always been my belief, and this is what I have always advised. We even had statistics: 5%-10% of Medicare patients are high risk and need a case manager, and 6%-10% of patients account for 85%-90% of costs. Although these figures may be accurate, the small percentage indicating patients who need a case manager is no longer accurate. The health-care system is fraught with pitfalls, bureaucracy, disinterest, and delay. Even straightforward care is no longer straightforward in its delivery. Case managers may not be the answer to the woes of the health-care system, but we are surely the patient's last hope for navigating the maze. PMID- 16720261 TI - Are you professionally and clinically competent? AB - Think about it-you graduated from medical school, a pharmacy, or nursing program and have been working in the world of health care free of books and tests for a number of years. There are no professors to keep you stimulated, no tests to ensure you keep up to date with the information required to complete your program, and no self-induced stress that pushes you to strive for that "A" to keep your grade point average up. So what is your motivation for keeping up with changes in today's fast-paced health-care world? PMID- 16720260 TI - Case managers play important part in employers' culture of health. AB - At Duncan Aviation, an aircraft support company based in Lincoln, Nebraska, meetings with the wellness staff are a job requirement. At least quarterly and often monthly, all managers and team leaders are required to schedule a presentation by the company's wellness staff as part of regular department or team meetings. "That way exposure to the wellness team and program is 100%, not just the 10% to 20% that we might have at a 'learn at lunch' program," explained wellness director Craig Johnson. "This is happening on company time. People are being paid to attend the presentation. It's part of the culture here." PMID- 16720262 TI - CE: "Medically futile" treatments require more than going to court. AB - With the unrelenting development of new medical technologies and increasingly more complex treatments, health-care providers sometimes find themselves faced with requests to provide treatments they believe to be medically futile. This language and resulting argument based on it can produce an adversarial posturing on the part of providers and families that frequently anticipates or threatens a legal solution. Although our legal system will choose sides on an issue, futility cases that have ended up in the courts have generally failed to definitively answer questions about how to address future dilemmas. A more helpful process is a clear procedure for addressing both sides of the question with the ultimate decision-making remaining within the health-care setting. The ethically appropriate solution lies within the context of a shared decision-making process between patient/family and physician/health-care provider that honors the values of both parties without assuming a unilateral decision-making stance. Case managers and direct-care providers, when faced with requests for treatments deemed to be medically inappropriate or futile, are challenged to understand and pursue this shared process. PMID- 16720263 TI - Bridging case management and primary care manager gaps in a military setting. AB - The team approach to case management (CM) has proven to be an effective method of providing quality outcomes, reducing fragmentation of care, improving communication, and reducing cost. Often CM teams consist of the patient, family/caregiver, physician, case manager, other health care personnel, clergy, home health agencies, employers, and health-plan administrators. This article focuses on the CM process implemented within a former TRICARE region to bridge the gap between the primary care manager (PCM) and CM. It discusses how the TRICARE Mid-Atlantic region identified and resolved barriers effecting collaboration between the PCM and CM. PMID- 16720264 TI - Speed dating for molecules CombinatoRx looks for that special synergy. PMID- 16720265 TI - New Ca2+ indicator has freedom to express. AB - In this issue of Chemistry & Biology, Tsien and colleagues describe a new family of highly sensitive genetically encoded calcium biosensors that are not affected by their local environment and can measure calcium concentration over a wide dynamic range. PMID- 16720266 TI - trans-Platinum reporting for duty. AB - HIV infects millions of people worldwide; however, no cures for this AIDS-causing retrovirus exist. A new approach using trans-platinum-nucleobase complexes to inhibit viral replication by disrupting the nucleocapsid protein-RNA interaction is presented in this issue of Chemistry & Biology. PMID- 16720267 TI - Activating hybrid modular interfaces in synthetic polyketide synthases by cassette replacement of ketosynthase domains. AB - Unnatural combinations of polyketide synthase modules often fail to make a polyketide product. The causes of these failures are likely complex and are not yet amenable to rational correction. One possible explanation is the inability of the ketosynthase (KS) domain to extend the ketide donated to it by the upstream module. We therefore addressed the problem by exchanging KS domains of the acceptor module in a combinatorial fashion and coexpressing these chimeric modules with ketide-donor modules that naturally interact with the transplanted KS. This approach was remarkably successful in activating previously unproductive bimodular combinations, and the results augur well for the ongoing development of molecular tools to design and produce novel polyketides. PMID- 16720268 TI - The gene cluster for fluorometabolite biosynthesis in Streptomyces cattleya: a thioesterase confers resistance to fluoroacetyl-coenzyme A. AB - A genomic library of Streptomyces cattleya was screened to isolate a gene cluster encoding enzymes responsible for the production of fluorine-containing metabolites. In addition to the previously described fluorinase FlA which catalyzes the formation of 5'-fluoro-5'-deoxyadenosine from S-adenosylmethionine and fluoride, 11 other putative open reading frames have been identified. Three of the proteins encoded by these genes have been characterized. FlB was determined to be the second enzyme in the pathway, catalyzing the phosphorolytic cleavage of 5'-fluoro-5'-deoxyadenosine to produce 5-fluoro-5-deoxy-D-ribose-1 phosphate. The enzyme FlI was found to be an S-adenosylhomocysteine hydrolase, which may act to relieve S-adenosylhomocysteine inhibition of the fluorinase. Finally, flK encodes a thioesterase which catalyzes the selective breakdown of fluoroacetyl-CoA but not acetyl-CoA, suggesting that it provides the producing strain with a mechanism for resistance to fluoroacetate. PMID- 16720269 TI - Cellular effects induced by the antitumor agent azinomycin B. AB - Studies on the mechanism of action of the antitumor agent azinomycin B in vitro suggest that the drug elicits its lethal effects by the formation of interstrand crosslinks within the major groove of DNA. Here, we demonstrate the biological effects of the drug in vivo. Fluorescence imaging revealed localization of azinomycin B in the nuclear region of yeast. Moreover, experiments with oligonucleotide microarrays examined the effects of the drug across the yeast transcriptome. The results demonstrated a robust DNA damage response that supports the proposed role of the drug as a covalent DNA modifying agent. RT-PCR analysis validated the gene changes, and flow cytometry of azinomycin-treated yeast cells demonstrated a phenotypic S phase shift consistent with transcriptional effects. PMID- 16720270 TI - A robust small-molecule microarray platform for screening cell lysates. AB - Herein we report the expanded functional group compatibility of small-molecule microarrays to include immobilization of primary alcohols, secondary alcohols, phenols, carboxylic acids, hydroxamic acids, thiols, and amines on a single slide surface. Small-molecule "diversity microarrays" containing nearly 10,000 known bioactive small molecules, natural products, and small molecules originating from several diversity-oriented syntheses were produced by using an isocyanate mediated covalent capture strategy. Selected printed bioactive compounds were detected with antibodies against compounds of interest. The new surface of the diversity microarrays is highly compatible with approaches involving cellular lysates. This feature has enabled a robust, optimized screening methodology using cellular lysates, allowing the detection of specific interactions with a broad range of binding affinity by using epitope-tagged or chimeric fluorescent proteins without prior purification. We believe that this expanded research capability has considerable promise in biology and medicine. PMID- 16720271 TI - Specific detection of glycans on a plasma membrane of living cells with atomic force microscopy. AB - Among the many alterations of cancer cells is the expression of different surface oligosaccharides. In this work, oligosaccharide expression in living cells (cancer and reference ones) was studied with atomic force microscopy by using lectins as probes. The unbinding force obtained for the same lectin type (concanavalin A or Sambucus nigra) suggested slightly dissimilar structures of binding sites of the same ligand type. For the lectin from Phaseolus vulgaris, a much larger unbinding force indicated a distinct structure of the binding site in cancer cells. The unbinding probability confirmed a higher content of both sialic acid and mannose-containing ligands in cancer and reference cells, respectively. These results demonstrate the potential of atomic force microscopy to directly probe the presence of molecules on a living cell surface, together with the quantitative description of their expression. PMID- 16720272 TI - Functional genomic analysis of alkaloid biosynthesis in Hyoscyamus niger reveals a cytochrome P450 involved in littorine rearrangement. AB - Tropane alkaloids are valuable pharmaceutical drugs derived from solanaceous plants such as Hyoscyamus niger (black henbane). The biosynthesis of these molecules, including the nature of the enigmatic rearrangement of (R)-littorine to (S)-hyoscyamine, is not completely understood. To test the hypothesis that a cytochrome P450 enzyme is involved in this rearrangement, we used virus-induced gene silencing to silence a cytochrome P450, CYP80F1, identified from H. niger roots by EST sequencing. Silencing CYP80F1 resulted in reduced hyoscyamine levels and the accumulation of littorine. Hyoscyamine was observed in CYP80F1-expressing tobacco hairy roots supplied with (R)-littorine. Expression in yeast confirmed that CYP80F1 catalyzes the oxidation of (R)-littorine with rearrangement to form hyoscyamine aldehyde, a putative precursor to hyoscyamine, and without rearrangement to form 3'-hydroxylittorine. Our data strongly support the involvement of CYP80F1 in the rearrangement of littorine to hyoscyamine. PMID- 16720273 TI - Ca2+ indicators based on computationally redesigned calmodulin-peptide pairs. AB - The binding interface of calmodulin and a calmodulin binding peptide were reengineered by computationally designing complementary bumps and holes. This redesign led to the development of sensitive and specific pairs of mutant proteins used to sense Ca(2+) in a second generation of genetically encoded Ca(2+) indicators (cameleons). These cameleons are no longer perturbed by large excesses of native calmodulin, and they display Ca(2+) sensitivities tuned over a 100-fold range (0.6-160 microM). Incorporation of circularly permuted Venus in place of Citrine results in a 3- to 5-fold increase in the dynamic range. These redesigned cameleons show significant improvements over previous versions in the ability to monitor Ca(2+) in the cytoplasm as well as distinct subcellular localizations, such as the plasma membrane of neurons and the mitochondria. PMID- 16720274 TI - Mimicking helical antibacterial peptides with nonpeptidic folding oligomers. AB - Unnatural oligomeric scaffolds designed to adopt defined secondary structures (e.g., helices), while retaining the chemical diversity of amino acid side chains, are of practical value to elaborate functional mimetics of bioactive alpha-polypeptides. Enantiopure N,N'-linked oligoureas as short as seven residues long have been previously shown to fold into a stable helical structure, stabilized by 12- and 14-membered H-bonded rings. We now report that eight residue oligoureas designed to mimic globally amphiphilic alpha-helical host defense peptides are effective against both gram-negative and gram-positive bacteria (including methicillin-resistant Staphylococcus aureus [MRSA]) and exhibit selectivity for bacterial versus mammalian cells. Circular dichroism (CD) spectroscopy studies suggest enhanced helical propensity of oligoureas in the presence of phospholipid vesicles. The utility of this new class of nonpeptidic foldamers for biological applications is highlighted by high resistance to proteolytic degradation. PMID- 16720275 TI - Targeting retroviral Zn finger-DNA interactions: a small-molecule approach using the electrophilic nature of trans-platinum-nucleobase compounds. AB - Noncovalent interactions are ubiquitous in ternary systems involving metal ions, DNA/RNA, and proteins and represent a structural motif for design of selective inhibitors of biological function. This contribution shows that small molecules containing platinated purine nucleobases mimic the natural DNA(RNA)-tryptophan recognition interaction of zinc finger peptides, specifically the C-terminal finger of HIV NCp7 protein. Interaction with platinum results in Zn ejection from the peptide accompanied by loss of tertiary structure. Targeting the NCp7-DNA interaction for drug design represents a conceptual advance over electrophiles designed for chemical attack on the zinc finger alone. These results demonstrate examples of a new platinum structural class targeting specific biological processes, distinct from the bifunctional DNA-DNA binding of cytotoxic agents like cisplatin. The results confirm the validity of a chemical biological approach for metallodrug design for selective ternary DNA(RNA)-protein interactions. PMID- 16720276 TI - Antagonists to human and mouse vascular endothelial growth factor receptor 2 generated by directed protein evolution in vitro. AB - Using directed in vitro protein evolution, we generated proteins that bound and antagonized the function of vascular endothelial growth factor receptor 2 (VEGFR2). Binders to human VEGFR2 (KDR) with 10-200 nM affinities were selected by using mRNA display from a library (10(13) variants) based on the tenth human fibronectin type III domain (10Fn3) scaffold. Subsequently, a single KDR binding clone (K(d) = 11 nM) was subjected to affinity maturation. This yielded improved KDR binding molecules with affinities ranging from 0.06 to 2 nM. Molecules with dual binding specificities (human/mouse) were also isolated by using both KDR and Flk-1 (mouse VEGFR2) as targets in selection. Proteins encoded by the selected clones bound VEGFR2-expressing cells and inhibited their VEGF-dependent proliferation. Our results demonstrate the potential of these inhibitors in the development of anti-angiogenesis therapeutics. PMID- 16720277 TI - Target range maximum of cyclosporine blood concentration two hours post dose in stable liver transplant patients. AB - Recently, single blood level measurement 2 hours after cyclosporine administration (C2) is taken as a more sensitive indicator of drug exposure in de novo transplant recipients than trough levels (C0). However, few studies focused on the determination of the C2 target range maximum and its associated adverse events in stable liver recipients. This prospective study was designed to assess the relative risk of developing CsA related side effects in patients with high C2 levels. Adverse effects were determined clinically, and by using a specially designed questionnaire. Eventual adverse events as well as C2 levels were determined repeatedly up to 4 times in 3-months intervals (observation period 9 +/- 3 months) in 36 long-term liver recipients (1-13.5 years post-transplant), in addition to conventional C0 levels. Cyclosporine dose was adjusted according to a predefined C0 target level range and clinical status. Totally 103 questionnaires and the corresponding paired CsA blood level records were obtained. C0 levels and C2 levels ranged from 90 to 287 (143 +/- 31) ng/ml and from 212 to 1358 (672 +/- 203) ng/ml respectively. No patient experienced a rejection episode during the observation period, demonstrating the efficiency of the immunosuppressive therapy. However, 33/36 patients (91%) showed symptoms attributable to CsA therapy. C2 levels above 750 ng/ml, determined at least twice in an interval of 3 months, were identified as a relevant risk factor for the presence of multiple adverse effects, which were defined as the combination of hypertension, renal insufficiency and more than two neurological complaints (RR = 3.11, p<0.01). This risk population was not completely identified by determination of C0 level. PMID- 16720278 TI - Assessment of body composition of patients with COPD. AB - The intention of the study was to determine nutritional state and body cell mass in patients with COPD in comparison with healthy volunteers between 50 and 75 years of age. Therefore, body cell mass (BCM), phase angle and the relation between extra cellular mass and body cell mass ECM/BCM was measured with the help of bioelectrical impedance analysis (BIA). While 10.4 % of COPD patients (male 5.0 %, female 18.8 %) had a BMI of < 18.5, no subject was underweight in the healthy volunteers. Overweight was found in 31.7 % patients (male 36.0 %, female 25.0 %) and in 54.2 % healthy subjects (male: 62.5 %, female 46.9 %), 17.1 % of our patients (male 16.0 %, female 18.8 %) versus 21.7 % of healthy subjects (male 14.3 %, female 28.1 %) were obese. While there was no statistic significance for a lower BMI in COPD patients, there were significantly decreased values concerning muscle mass -- represented by BCM-values -- and state of nutrition -- represented by BCM, phase angle and ECM/BCM values -- in COPD patients compared with healthy volunteers. These results suggest that BMI alone doesn't allow conclusions regarding to nutritional state and physical training. A malnutrition requiring intervention might exist in spite of a normal or even high BMI in COPD patients. PMID- 16720280 TI - Modulation of uterine contractility and peristalsis by oxytocin in estrogen primed non-pregnant swine uteri. AB - Oxytocin is one of the most potent uterotonic agents and is known to fluctuate throughout the menstrual cycle, showing an increase during sexual stimulation and arousal, with a peak during orgasm in women. To date, limited data are available on the effects of oxytocin on the regulation of uterine contractility and transport mechanisms in human reproduction. The goal of this study was to evaluate the effects of oxytocin on uterine contractility and peristalsis in estrogen-primed non-pregnant uteri. In an extracorporeal perfusion model of the swine uterus the effect of dynamic changes in uterine contractility and peristalsis in response to oxytocin and estrogen administration was observed. Spontaneous uterine contractility and oxytocin-induced uterine contractility and peristalsis with and without estrogen perfusion were assessed using an intrauterine double-chip microcatheter. Spontaneous peristalsis and oxytocin induced contraction waves without estrogen perfusion resulted in a slightly higher intrauterine pressure in the isthmus uteri in comparison with the corpus uteri, while the peristaltic waves were seen to start mostly in the corpus uteri, moving in the direction of the cervix. While after estrogen perfusion oxytocin produced a significant increase in intrauterine pressure in the isthmus uteri compared to the corpus uteri, and 80% of the peristaltic waves started in the isthmus uteri, moving in the direction of the corpus uteri. This observation strengthens the view that oxytocin is able to support directed transport mechanism in the female genital tract only in the presence of estrogens. The biological role of oxytocin increase during sexual stimulation and arousal with a peak during orgasm for the mechanisms of reproduction may be to stimulate directed uterine transport mechanisms in the presence of estrogens. PMID- 16720279 TI - Unilateral generalized morphea is a rare variant of localized scleroderma. AB - Localized scleroderma (LS) is a rare connective tissue disorder generally involving the entire dermis and usually limited to the subcutaneous tissue. However, it may progress to large indurated plaques, growth retardation, muscle atrophy, and even to flexion deformities or poorly healing ulcerations. LS has been classified as plaque, generalized, bullous, linear, and deep forms exhibiting different clinical subtypes. Recently, an unusual case of unilateral generalized morphea (UGM) in childhood extending from the middle dermis to the subcutaneous fat tissue has been reported. We here describe four young patients exhibiting a similar subtype of LS. All patients demonstrated a prominent unilateral skin involvement starting in childhood or adolescence. Histology revealed prominent accentuation of intradermal involvement. Except for positive anti-nuclear antibodies, no specific antibody pattern could be observed. In presenting these clinically homogenous cases we hereby introduce UGM as an extreme variant of the linear form of LS in childhood. As the onset of UGM usually occurs in pediatric patients, pediatricians should be cognizant of the presentation of this uncommon condition. Treatment with combined low-dose methotrexate and pulsed high-dose corticosteroid therapy might represent a promising treatment option for UGM. PMID- 16720281 TI - Disseminated mycobacterium bovis infection in an immunocompetent host. AB - We report about a rare case of disseminated Mycobacterium bovis infection in a 61 year old female immunocompetent patient with involvement of the lung, the brain, the spleen and spine. The patient had intracerebral tuberculomas with paradoxical enlargement during the first weeks of therapy. We reviewed the data of our microbiological department and found five other patients with Mycobacterium bovis infection diagnosed between 1999 and 2004, which are 5.8 % of all diagnoses of tuberculosis during this period. PMID- 16720282 TI - Aggressive angiomyxoma of the prostate mimicking benign prostatic hyperplasia. AB - Aggressive angiomyxoma (AAM) is a locally invasive soft tissue tumor with a high risk of local recurrence but without metastatic spread. The mesenchymal tumor is relatively site-specific and has a peak incidence in females in their 2nd or 4th decade. Only few cases in males have been reported in the literature. We describe what we think is the first case of an aggressive angiomyxoma arising in the prostate presenting with classical symptoms of benign prostatic hyperplasia. PMID- 16720283 TI - Gluteal compartment syndrome after prostatectomy caused by incorrect positioning. AB - Gluteal compartment syndrome is an uncommon and rare disease. Most reasonable causes for the development of this disease are trauma, drug induced coma, Ehlers Danlos syndrome, sickle cell associated muscle infarction, incorrect positioning during surgical procedures and prolonged pressure in patients with altered consciousness levels. The diagnosis requires a high index of suspicion, especially in postoperative patient where sedation or peridural anaesthesia can confound the neurological examination. Early signs include gluteal tenderness, decrease in vibratory sensation during clinical examination and increasing CK in laboratory findings. We present a case of a 52 year-old patient, who developed gluteal compartment syndrome after radical prostatectomy in lithotomic position. After operation, diuresis decreased [<50 ml/h] and CK [93927 U/l], LDH [1528 U/l], creatinin [1.5 mg/dl] and urea [20 mg/dl] increased in laboratory findings. Despite peridural anaesthesia, the patient complained about increasing pain in the gluteal region and both thighs. His thighs and the gluteal region were swollen. Passive stretch of the thighs caused enormous pain. The compartment pressure was 92 mmHg. Therefore, emergency fasciotomy was performed successfully. The gluteal compartment syndrome was most likely caused by elevated pressure on the gluteal muscle during operation. We suggest heightened awareness of positioning the patient on the operating table is important especially in obese patients with lengthy operating procedures. PMID- 16720284 TI - Benign symmetric lipomatosis (Launois-Bensaude syndrome). A rare cause of muscular weakness. AB - A 76-year-old female patient is presented who suffered from muscular weakness in arms and legs. She was obese and had a symmetric accumulation of fatty tissue with a bumpy structure at both arms which gave the patient a pseudoathletic appearance. Fatty tissue accumulations were present at both shoulders, arms, at both thighs, at the back and the abdomen. She suffered from benign symmetric lipomatosis (BSL), also called Launois-Bensaude syndrome (LBS), which is a rare disorder of unknown origin and poorly understood pathophysiology. It is believed to be a disease of disturbed lipogenesis induced by catecholamines. The syndrome is often associated with features of metabolic syndrome such as diabetes mellitus, hyperuricemia, hyperlipidemia and hypertension and is associated with polyneuropathy which is an integral part of the disease. Therapeutic options are pharmacological treatment with salbutamol and surgical procedures such as lipectomy or liposuction. PMID- 16720285 TI - Dosing pattern of renal replacement therapy in acute renal failure: current status and future directions. AB - Acute renal failure (ARF) is associated with poor clinical outcome, particularly in critically ill patients. Different renal replacement techniques are available for treatment of ARF, but a general consensus of standardization of treatment dose is lacking. Recent prospective investigations in critically ill patients with ARF utilizing either intermittent or continuous renal replacement therapy established a clear dose-outcome relationship. However, several surveys of current practices of renal replacement therapy dosing in acute renal failure revealed, that the majority of ICU patients with ARF received dialysis doses lower than prescribed for stable end-stage renal disease patients. Most nephrologists did not perform formal dose calculation. Future patterns of dosing renal replacement therapy should focus both on an earlier start and a higher dose to reduce the excess mortality associated with ARF in critically ill patients. PMID- 16720286 TI - Comparing the effect of ATRA, 4-HPR, and CD437 in bladder cancer cells. AB - Clinical trials have explored the use of natural and synthetic retinoids for the prevention of bladder cancer recurrence. Natural retinoids have been shown to inhibit bladder cancer growth. Here, we compared the effects of natural and synthetic retinoids in bladder cancer cells. Bladder cancer cell lines were treated with all-trans-retinoid acid (ATRA), N-4-hydroxyphenyl-retinamide (4-HPR) and 6-[3-(1-adamantyl)-4 hydroxyphenyl]-2-naphthalene carboxylic acid (CD437). Their effects on cell growth, apoptosis, cell cycle, gene expression, and retinoid acid receptors (RARs) and the JWA-retinoid response gene were assessed. Most of the bladder cancer cells were resistant to ATRA (1 and 10 microM). 4-HPR inhibited cell growth by 90% at 10 microM; however, CD437 showed the same effect at 1 microM. 4-HPR and CD437 increased G1 and decreased S phase. The three retinoids differentially affected p53, RARs, and JWA. Only CD437 increased Caspase 3 expression. The results demonstrated that 4-HPR and CD437 were more potent growth inhibitors and apoptosis inducers than ATRA. However, 4-HPR was effective at a concentration at least 10 microM. The in vitro results suggested the higher dose of 4-HPR in chemoprevention trial be considered. PMID- 16720287 TI - Subcellular structures of mycoplasmas. AB - Although the field of prokaryotic cell biology is well-advanced now, mycoplasmas were the first bacteria in which the existence of a cytoskeleton was postulated. Despite this head-start, the cytoskeletons of mycoplasmas are presently less well understood than those of other bacteria. This deficit is principally attributable to three factors: the novel nature of most of the cytoskeletal elements as compared with other bacteria, which have the advantage of being related to eukaryotic cytoskeletal proteins; differences among the cytoskeletons of different mycoplasma species; and the fastidiousness of mycoplasmas, which complicates efforts to perform protein biochemistry. In better studied mycoplasmas like Mycoplasma pneumoniae, a major component of the cytoskeleton is associated with the attachment organelle, a polar structure that is essential for adherence to host cells, involved in gliding motility, and associated with cell division. Mycoplasma mobile also has structures that appear to be involved in gliding motility, though in contrast to the structures of M. pneumoniae, these are extracellular. Some other species also have distinct subcellular structures. PMID- 16720288 TI - Mycoplasma hominis and Trichomonas vaginalis: a unique case of symbiotic relationship between two obligate human parasites. AB - Mollicutes are the smallest and simplest self-replicating microorganisms. Despite the minimal genome and apparent lack of complexity, mycoplasmas show a high degree of adaptation to the most diverse environments. Mycoplasma hominis is a human sexually transmitted mycoplasma which is able to establish a biological association with Trichomonas vaginalis, a pathogenic flagellated protist. M. hominis and T. vaginalis share the same specific natural niche, the human genitourinary tract. Symbiotic relationships between unicellular eukaryotes and bacteria are well known and have been extensively studied, providing interesting insights into the biology of one or both the symbionts. The relationship between T. vaginalis and M. hominis is unique in that it was the first described association of two obligated human parasites. Several aspects of this relationship have been investigated, showing how the trichomonad may be viewed not only as a new niche for M. hominis, but also as a "Trojan horse" for the transmission of the bacterial infection to the human host. PMID- 16720289 TI - Effect of pH-variation on insertion and ion channel formation of human calcitonin into planar lipid bilayers. AB - Human calcitonin is the physiological hormone involved in calcium-phosphorus homeostasis. However, its use is limited by its propensity to form aggregates. We find that the type of host lipid has a pronounced influence on human calcitonin fibrillation or incorporation, as assessed by channel formation, in planar lipid membranes at neutral pH. At pH 7, human calcitonin is able to interact and form channels with negatively charged dioleoyl-phosphatidylglycerol (DOPG) bilayers and with zwitterionic palmitoyl-oleoyl phosphatidylcholine (POPC) bilayers containing 15% negatively-charged DOPG, but not with POPC bilayers. At low pH (4.5 and 3.8), the conformational variation of the peptide enables it to insert into POPC and POPC:DOPG but not into DOPG bilayers. The model proposed for human calcitonin interaction and channel formation at acidic pH was based on theoretical predictions of the protonation-deprotonation state of some amino acids, in particular in the fibrillating sequence of peptide molecules; the length of the alpha-helix, and the electrostatic and/or hydrophobic interaction also seem to be relevant. These results may suggest that human calcitonin at low local pH could be involved in osteoclasts' calcium-sensitive permeability through channel formation and/or receptor interaction. PMID- 16720290 TI - Intravesical N-(4-hydroxyphenyl) retinamide and adriamycin induces apoptosis in bladder cancer. AB - The objective of this study was to evaluate the intravesical application of N-(4 hydroxyphenyl) retinamide (4-HPR) and adriamycin (ADM), as a treatment modality in a an animal model of chemically-induced bladder cancer. Bladder cancer developed in 50.0% of female Wistar rats, 4-6 weeks after intravesical application of the chemical carcinogen, N-methyl-N-nitrosourea (MNU). There was no significant difference in side effects induced by local versus systemic 4-HPR. Although tumor growth was inhibited by 4-HPR and ADM alone, tumor size was lower when both agents were used together. Apoptosis occurred at a higher rate in the combination group than when 4-HPR or ADM was used alone. The results suggest that intravesical use of 4-HPR and ADM may increase their efficacy in treatment of bladder cancer. PMID- 16720292 TI - Gene therapy of focal cerebral ischemia using defective recombinant adeno associated virus vectors. AB - This review presents our experience and results concerning cerebral stroke gene therapy with a rat model subjected to rAAV-vector delivered IL-1ra and GDNF. The methodology involving the production of high-titer recombinant adeno-associated virus vectors in the absence of helper adenovirus and the creation of a tri vessel ligation model of focal ischemic cerebral stroke in rats are described in detail. Furthermore, a literature review of other viral vectors, murine models of focal cerebral ischemia and candidates for therapeutic transgenes used for cerebral stroke gene therapy are presented. Lastly, the potentials and limitations of stroke gene therapy are discussed adding an analysis of possibilities of future experiment designs. PMID- 16720291 TI - Polymorphisms of metabolic enzyme genes, living habits and prostate cancer susceptibility. AB - In this report, genetic polymorphism of phase I and II metabolic enzyme (CYP2E1, CYP17, GSTM1 and GSTT1) genes, living habits, and risk of prostate cancer (PCa) was studied in 163 patients with prostate carcinoma of Han nationality in Southern China and 202 age-matched controls. The genotypic polymorphism of CYP2E1, CYP17, GSTM1 and GSTT1 genes was analyzed by PCR-RFLP assay using genomic DNA isolated from peripheral blood lymphocytes. The significant risk factors for PCa included long-term exposure to toxicant (OR=2.27, 95%CI: 1.26-4.09), the tumor history of lineal consanguinity (OR=2.19, 95%CI: 1.30-3.67), sexual history before age 30 of no more than 8 times per month (OR=1.85, 95%CI: 1.22-2.81), deep inhalation of cigarette smoke (OR=2.01, 95%CI: 1.20-3.37) or heavy smoking (OR=1.67,95%CI: 1.01-2.76). Among individuals with long-term heavy smoking without tea-drinking habit, the risk increased significantly (OR=4.27, 95%CI: 1.62-11.24 and OR), 2.76, 95%CI: 1.20-6.32). CYP2E1 C1/C1 genotype significantly increased the risk for PCa (OR=1.61, 95%CI: 1.04-2.49) with an apparent interaction with alcohol (OR=2.07, 95%CI: 1.07-4.00). However, stratification by the amount of accumulative smoking revealed that among people with a heavy smoking history, the individuals with the CYP2E1 C1/C1 genotype (OR=2.55, 95%CI: 1.20-5.43) and the individuals with GSTT1 null genotype (OR=2.23, 95%CI: 1.09 4.57) showed a significantly increased risk. Any other significant results with GSTM1 or CYP17 genes were not observed in this research. Individuals with more sensitive genotypes (from one to four) were at an increased risk. The data show that, in the development of PCa, there are many interactions among predisposing genotypes and genetic polymorphisms and unhealthy living habits. Individuals with more susceptible genotypes and unhealthy habits such as prolonged exposure to smoking are at an increased risk. PMID- 16720293 TI - Significance of erb-B2 immunoreactivity in cervical cancer. AB - C-erbB2 is over-expressed or amplified in many carcinomas. We assessed the relationship between erb-B2 immunoreactivity, and its predictive role in progression-free survival and treatment outcome in patients with cervical carcinoma. Sections from 65 cervical carcinoma were immunostained with antibody to p185 erbB2. Immunoreactive ErbB2 was found in 25 patients (38%) [+ 15 pts. (23%); ++ 10 pts. (15%)]. There were no correlation with age, performance status, grading and histology. Erb-B2 immunoreactivity significantly correlated with stage of the disease. Positive immunoreactivity was found in 63%, 44%, 14% and 0% of stage I, II, III and IV carcinomas, (p = 0.0045). Progression-free survival was longer in erb-B2 positive patients without reaching significance. No correlation was found between erbB2 and response to radiotherapy or chemotherapy. In conclusion, a significant proportion of stage I and II cervical cancer express erb-B2 compared to more advanced stages. Expression of the oncogene does not appear to be related to prognosis or treatment outcome. PMID- 16720294 TI - Women with HNPCC: a target population for the chemoprevention of gynecologic cancers. AB - Hereditary nonpolyposis colorectal cancer syndrome (HNPCC) is an autosomal dominant inherited cancer susceptibility syndrome that is associated with increased risk of development of endometrial and ovarian cancers. Therefore, women with HNPCC are candidates for chemoprevention of gynecological cancers. Although there have been a number of clinical trials examining chemoprevention strategies for colorectal cancer in HNPCC, the information regarding gynecological cancers in HNPCC remains limited. The purpose of this review is to provide an analytical background about the current knowledge regarding gynecological cancers in patients with HNPCC. PMID- 16720295 TI - From existing therapies to novel targets: a current view on melanoma. AB - Identifying new drugs and targets for melanoma therapy is critical, considering that melanoma, the most dangerous form of skin cancer, is resistant to currently available therapeutics. Much work has been focused on finding novel drugs and exploring different treatment options that could increase the overall survival of patients. In our laboratory we have developed mouse models to study melanoma. We discovered that aberrant expression of metabotropic glutamate receptor 1 (Grm1) in melanocytes promotes melanoma development in vivo. Grm1 is a seven transmembrane domain G-protein coupled receptor that is normally expressed and functional in the central nervous system. The natural ligand of Grm1 is glutamate. Signaling by the major neurotransmitter glutamate has been well characterized in neuronal cells; however glutamate signaling in other tissues is not well understood. We demonstrated that Grm1 signaling in melanoma cells is mediated by the Ras/Raf/MEK/ERK pathway, one of the major pathways previously shown to be activated in human melanoma cells. Based on these earlier studies and results from our recent work, we predict that inhibition of Grm1 signaling and its downstream cascade may potentially provide new, effective therapies for melanoma patients. In this review, we propose several attractive targets. PMID- 16720296 TI - Secreted WNT antagonists as tumor suppressors: pro and con. AB - Dysregulation of Wnt signaling is common in a variety of human malignancies. Activation of the canonical Wnt or beta-catenin pathway has been especially well documented in cancer, although other non-canonical Wnt signaling pathways also have been implicated in neoplasia. In most instances, constitutive signaling through the beta-catenin pathway involves activation of effector molecules or loss of tumor suppressor function downstream of Wnt binding to its cell surface receptors. Nonetheless, in recent years increasing evidence suggests that secreted Wnt antagonists act as tumor suppressors, with their expression often silenced by promoter hypermethylation. This implies that maximal constitutive signaling in cancer requires unimpaired Wnt stimulation at the cell surface as well as enhanced signal propagation within the cell. However, an understanding of the role secreted Wnt antagonists may play in cancer is complicated by the multiplicity of these proteins, their potential Wnt-independent activities and observations indicating that sometimes they may promote tumor growth. Just as the particular function of Wnt signaling in development and homeostasis varies with the setting, the impact of secreted Wnt antagonists on neoplasia depends on the molecular, cellular and tissue context. PMID- 16720297 TI - WNT signaling in malignant mesothelioma. AB - Neoplastic transformation of mesothelium is commonly associated with exposure to asbestos and gives rise to malignant mesothelioma, an aggressive disease that has proved particularly refractory to conventional anti-cancer therapies. The Wnt signaling pathways play key roles in fundamental processes, which include both development and homeostasis. The importance of these pathways in tumorigenesis is emphasized by the many cancers which show aberrations in Wnt signaling. In this review we examine the current evidence for activation of Wnt signaling and the abnormal expression of specific molecules in malignant mesothelioma. PMID- 16720298 TI - Development of ELISA for metallothionein-II allows determination of heavy metal pollution of fresh water. AB - Metallothionein (MT), a metal-binding protein induced primarily by heavy metals in vertebrates, is considered a biomarker for environmental heavy-metal contamination. To investigate heavy metal pollution in the freshwater environment, MT-I and MT-II were purified from livers of crucian carp (Carassius carassius) by gel exclusion chromatography and ion exchange chromatography. To detect the purified MT-II, a specific monoclonal antibody (mAb) against crucian carp MT-II was produced from the hybridoma strains by cell-cell fusion. By using Enzyme-Linked Immunosorbent Assay (ELISA) with this mAb, the purified crucian carp MT-II was detected with a high specificity and sensitivity. There was a good correlation between the amount of MT-II in carp livers and the concentration of heavy metals in water. ELISA was then used to evaluated the degree of heavy metal pollution in two freshwater systems. The results indicate that the MT-II content in carp liver tissue can be used as an indicator of environmental heavy-metal pollution. PMID- 16720299 TI - Genes regulating implantation and fetal development: a focus on mouse knockout models. AB - Timely and efficient regulation of blastocyst implantation and fetal growth are essential for the successful reproduction of viviparous mammals. Disruptions in this regulation can result in a wide variety of human gestational complications including infertility, spontaneous abortion, fetal growth restriction, and premature delivery. The role of several groups of factors, including cytokines, hormones, transcription factors, extra-cellular proteinases, and angiogenic factors has been suggested in both implantation and regulation of fetal growth. Due to the inherent difficulties of studying implantation and fetal development in humans, much of our knowledge of the genes involved in these processes has been derived from animal models. The critical genetic loci involved in blastocyst implantation and fetal growth will be discussed with a focus on those genes with available mouse knockout models. PMID- 16720300 TI - The tyrosine phosphatase HD-PTP is regulated by FGF-2 through proteasome degradation. AB - Angiogenesis is essential in development and wound healing and contributes to the pathogenesis of many diseases. The signalling pathways activated in angiogenesis are, in part ,known and the overall tyrosine phosphorylation of cellular proteins plays a relevant role. By RNA fingerprinting, we isolated a tyrosine phosphatase, HD-PTP, modulated in human endothelial cells exposed to human immunodeficiency virus type 1 Tat, a viral protein known to be angiogenic. For the first time, we describe HD-PTP at the protein level. HD-PTP, a 185 kDa cytosolic protein which is expressed in endothelial cells of different origin. We show that HD-PTP is upregulated by Tat at the mRNA but not at the protein level. HD-PTP protein is differentially modulated by two angiogenic growth factors. While Vascular Endothelial Growth Factor does not affect protein levels, Fibroblast Growth Factor-2 induces HD-PTP degradation via the proteasome system. PMID- 16720301 TI - Signaling events during male germ cell differentiation: update, 2006. AB - The intracellular transduction of exogenous and cell-autonomous stimuli triggers the transformation of a multipotent stem cell, the spermatogonion, into a highly differentiated, motile and fertile cell, the spermatozoon. This differentiation process is mediated by cell-cell contact and via key players including hormones, growth factors, and cytokines. Female hormones, estrogens and progestins, play a role in the production and functionality of spermatozoon. New findings, however, reconsider the direct action for estrogens on male germ cells while progestins work through non-canonical receptors. Similarly, testosterone, the male hormone, besides acting through its receptor expressed in the somatic cells of testis, seems to work by means of non-classical mechanisms. The recent identification of growth factors, transcriptional regulators, and media for in vitro growth of spermatogonial stem cells should now make it feasible to unravel the entire spermatogenic process. A peculiar feature of the meiotic cycle is the maintenance of condensed chromatin so that DNA duplication is prevented and reduction of genome is achieved. Recently, molecular mechanisms that lead to such a condensation have been discovered. Junctional intercellular complexes between Sertoli and germ cells are critical for coordinating spermatogenesis. Molecular players involved in such cell-cell communication have been identified in Sertoli cells. Now, there is also a need for unravelling the germ cell molecules involved. These issues are the major topics which are discussed here with the goal to suggest a possible answer. PMID- 16720302 TI - Cutaneous photoprotection and melanoma susceptibility: reaching beyond melanin content to the frontiers of DNA repair. AB - Melanoma is the most devastating form of skin cancer. The steady increase in the incidence of melanoma, its resistance to chemotherapy, together with its high potential to metastasize, have emphasized the importance of its prevention. It is becoming clear that solar ultraviolet radiation is a main culprit in the etiology of melanoma, the same as in basal and squamous cell carcinomas. It is commonly accepted that skin pigmentation and melanin content are principal determinants of the susceptibility to melanoma and other sun-induced skin cancers. Although this is generally true, however, prediction of melanoma risk based solely on pigmentary phenotype is not always precise and fails to identify high-risk individuals with dark skin color. Other important risk factors need to be considered and better defined, particularly DNA repair capacity. Emerging studies have revealed the role of melanoma susceptibility genes in regulating DNA repair, and indicated that melanoma patients have a lower DNA repair capacity than the general population. As the response of human melanocytes to ultraviolet radiation is modulated by an array of paracrine factors, we have focused our investigation on the role of melanocortins and the melanocortin 1 receptor, as well as endothelin-1, in this response. We have discovered novel roles for melanocortins and endothelin-1 as survival factors that rescue human melanocytes from ultraviolet radiation-induced apoptosis, and importantly enhance repair of DNA photoproducts and reduce the release of hydrogen peroxide that can cause oxidative stress. Our findings, together with epidemiological data showing that loss-of-function mutations in the melanocortin-1 receptor gene increase the risk of melanoma, substantiate the role of DNA repair in melanoma genesis, and suggest that responsiveness to melanocortins and endothelin-1 is important for melanoma prevention. PMID- 16720303 TI - Effect of straining on the muscles of the anterior abdominal wall. Identification of the 'straining-abdominal wall reflex'. AB - The external and internal oblique, transverse abdominis and the rectus abdominis muscles constitute the anterolateral abdominal wall muscles. They are striated and contract voluntarily. We investigated the hypothesis that contraction of these muscles by coughing or straining, can also occur as a reflex. Effect of straining on muscles was tested in 19 healthy volunteers. The intra-abdominal pressure was measured by a manometric catheter introduced into the rectum. The response of the muscles to straining was recorded by a needle electrode inserted into each of the muscles. Similar to voluntary contractions, sudden and sustained straining produced increase in the rectal pressure and the motor unit action potentials of each of the muscles which was abolished by anesthesia. These findings suggest presence of a 'straining-abdominal wall reflex'. PMID- 16720304 TI - Stress-induced hyperalgesia: animal models and putative mechanisms. AB - Stress has been shown to affect brain activity and promote long-term changes in multiple neural systems. A variety of environmental and/or stressful stimuli have been shown to produce analgesia, a phenomenon often referred to as stress-induced analgesia (SIA). However, acute and chronic stresses also produce hyperalgesia in various behavioral tests. There are now several animal models in which stress enhances nociceptive responses. The dysfunction of the hypothalamo-pituitary adrenocortical axis (HPA axis) and multiple neurotransmitter systems in the central nervous system (CNS), including endogenous opioid, serotonergic and noradrenergic systems, has been reported. These stress-induced hyperalgesia models may contribute to a better understanding of chronic pain and provide a more rational basis for drug therapies in a variety of pain syndromes. PMID- 16720305 TI - Degradation of securin in mouse and pig oocytes is dependent on ubiquitin proteasome pathway and is required for proteolysis of the cohesion subunit, Rec8, at the metaphase-to-anaphase transition. AB - Although securin/separase/cohesion pathway was reported to regulate chromosome segregation during meiotic metaphase-to-anaphase transition, little biochemical evidence was provided. We recently found that oocytes could not progress beyond meiotic metaphase when ubiquitin-proteasome pathway was inhibited, but the mechanisms remain unclear. In the present study, we investigated the quantity of securin and Rec8 protein and the localization of securin, a cohesion subunit, during oocyte meiosis providing data in support of the hypothesis that the effect of ubiquitin-proteasome pathway on metaphase-to-anaphase transition was mediated by regulating securin and Rec8 degradation in mouse and pig oocytes. In germinal vesicle-stage oocytes, immunostaining of securin was mainly localized in the germinal vesicle. Shortly after germinal vesicle breakdown, immunoreactive securin accumulated around the condensed chromosomes at prometaphase I. At metaphase I and metaphase II, when chromosomes were organized at the equatorial plate, immunoreactive securin was concentrated around the aligned chromosomes, putatively associated with the position of the metaphase spindle. The accumulation of securin could not be detected at anaphase I and anaphase II. In both mouse and pig oocytes, Western blot analysis showed that securin protein was low at germinal vesicle stage, reached the highest level at metaphase I, while decreased at anaphase I. Securin was increased again at metaphase II, while it was decreased at anaphase II. Rec8 protein was present in germinal vesicle-stage oocytes and remained until metaphase I, while it was decreased at anaphase I. Like securin, Rec8 was increased at metaphase II, while it was decreased again at anaphase II. The inhibition of the ubiquitin-proteasome pathway inhibited the decrease in securin and Rec8 at metaphase-to-anaphase transitions in both mouse and pig oocytes. Microinjection of securin antibody into MII-arrested oocytes leads to the degradation of Rec8. In conclusion, these results suggest that the proteolysis of securin is dependent on ubiquitin-proteasome pathway and is necessary for the degradation of Rec8 during meiotic metaphase-to-anaphase transitions in mouse and pig oocytes. PMID- 16720306 TI - The role of laparoscopy in pancreatic surgery. AB - Carcinoma of the pancreas is extremely common, with a five-year mortality rate of about 95-99%. Radical surgery requires good technical skill and can cause complications and operative mortality, but should be avoided in patients with extrapancreatic involvement. Advances in dynamic spiral CT-scan have decreased the number of unnecessary laparotomies. VLS is indicated in cases of pancreatic mass deemed resectable or "doubtful" by CT-scan. Direct laparoscopic visualization can be combined with intraoperative laparoscopic ultrasonography (LUS), which has shown a positive predictive value of resectability of 91%. Laparoscopic pancreatoduodenectomy (LPD) shows a high rate of complications and should be performed by very well-trained surgeons. Laparoscopic distal pancreatectomy (LDP) with an "en bloc" splenectomy and spleen preservation should be performed. PMID- 16720307 TI - Surgical treatment of differentiated thyroid carcinoma: a retrospective study. AB - A retrospective study was carried out to assessed reliability of the prognostic factors (histology, age, sex, and stage), and standard procedures for the surgical treatment of differentiated thyroid cancers (DTC). From the 144 DTC cases reviewed with follow-up ranging from 1 to 25 years (m = 6.33 years), total mortality for cancer was found to be 55% (8 patients), with a predictive positive value for recurrence of 95.4% and 91.8% at 12 and 24 months, respectively. Median survival was 8.8 years (range 1 to 25 years). The multivariate analysis showed that factors such as age > 45 years, histology of intermediate malignancy, size up to 1.5 cm, and presence of metastases, significantly worsened the prognosis, regardless of the intervention that was carried out. We suggest total thyroidectomy for the treatment of benign pathologies and confirmed or suspected cases of cancer. We reserve loboisthmectomy for the treatment of benign pathologies confined to one lobe or those with FNAB suggesting a follicular neoplasm. PMID- 16720308 TI - The two helicases of herpes simplex virus type 1 (HSV-1). AB - Herpes simplex virus type 1 (HSV-1) encodes two helicases both of which are essential for viral DNA synthesis. UL9 binds specifically to the origins of replication and is believed to initiate DNA replication at one of three origins of replication located in the HSV-1 genome. The heterotrimeric helicase-primase complex, encoded by the UL5, UL8 and UL52 genes, is believed to unwind duplex viral DNA at replication forks and to prime lagging strand synthesis. Functional analyses of UL9 and the helicase-primase complex will be discussed with attention to the roles these proteins play during HSV-1 replication. PMID- 16720309 TI - Real-time contrast-enhanced specific ultrasound in staging and follow-up of splenic lymphomas. AB - From January 2003 to April 2005 we studied 25 lymphoma patients (10 with HD, 4 with low-grade NHL, 6 with high-grade NHL and 5 with chronic lymphatic leukaemia; 14 men, 11 women, age range 28-79 years). After a baseline US study we rapidly injected 4.8 mL of the second-generation microbubble contrast agent SonoVue (Bracco, Italy). Contrast enhanced studies were carried out with the contrast specific software named Contrast Tuned Imaging (Esaote, Italy) using a continuous, harmonic acquisition and a low acoustic pressure. The CS-US findings were correlated with results of standard tools, including CT, MRI, US follow up. CS-US revealed correctly 47 out of the 52 lesions identified by CT scan, in the absence of false positive findings (sensitivity = 90%; Specificity = 100%, in comparison to CT scan). Complete concordance in evaluating the lesion extension of the CS-US in respect to CT was 88%, while underestimate occurred in 9% and overestimate in 3% of cases. On the contrary, basic sonography defined correctly the dimensional alteration in 52% of the cases, underestimated in 35% and overestimated in 13%, thus showing significantly lower accuracy (chi-square = 30.0, p < 0.001). In our experience, CS-US was superior to conventional sonography even from a qualitative point of view. PMID- 16720311 TI - Chemoprevention of breast cancer: current status and future prospects. AB - Chemoprevention plays an important role in the prevention of cancer. Due to being an estrogen-dependent cancer, breast carcinomas are ideal candidate for chemoprevention. The two main approaches to chemoprevention of breast cancer are to attain a balance in estrogen or to eliminate all estrogens by selective estrogen receptor modulators (SERMs) and aromatase inhibitors (AIs). A series of clinical trials have proven tamoxifene quite useful as a preventive strategy for women at high risk of breast cancer. The third-generation, AIs, hold a great promise in chemoprevention of breast cancer. In this review, the mechanisms, side effects and main clinical trials of SERMs and AIs are discussed. The use of other drugs including CoQ10, retinoids, cyclooxygenase (COX)-2 inhibitors, vitamin E, soy isoflavones, tea polyphenols and statins in chemoprevention is introduced. Since most studies on these drugs were conducted on animal models or cell culture, further studies are needed to determine the efficacy of these drugs in chemoprevention of human cancers. PMID- 16720312 TI - Perinatal invasive malignant diseases: a review of twenty-five cases in South China. AB - Malignant neoplastic diseases (MND) are unusual complications during perinatal period and compose a dilemma for both patients and the health practitioners. Little is known about the information in Chinese suffering perinatal MND. Analyzing medical records and questionnaire, information on a series of 25 patients with a diagnosis of perinatal MND was collected from 3 medical centers between 1992 and 2004. Among all the 25 patients, 10 selected termination of the pregnancies and the other 15 continued their pregnancies until labor voluntarily, both groups obtain anti-malignancies therapies during the perinatal period. The two groups were not statistically different for the age of pregnancy, gravid and parity number, interval weeks between symptoms emergence and diagnosis of invasive malignant disease, as well as the occurrence rates of major side effects induced by malignant therapies. No statistical differences in overall survival and disease-free survival between the two groups, including the age of pregnancy, gravid and parity number, obstetric bleeding rates, neonatal distress rates, neonatal weight and puerperal morbidity rates. The patients' neonates all show no serious complications. In conclusion, pregnancy may not affect the course of MND, and termination of pregnancy may not benefit the maternal-fetal conditions, in a macroscopical point of view. PMID- 16720310 TI - Corticotropin releasing hormone and the skin. AB - Cotricotropin-releasing hormone (CRH) and related peptides are produced in skin that is dependent on species and anatomical location. Local peptide production is regulated by ultraviolet radiation (UVR), glucocorticoids and phase of the hair cycle. The skin also expresses the corresponding receptors (CRH-R1 and CRH-R2), with CRH-R1 being the major receptor in humans. CRH-R1 is expressed in epidermal and dermal compartments, and CRH-R2 predominantly in dermal structures. The gene coding for CRH-R1 generates multiple isoforms through a process modulated by UVR, cyclic adenosine monophosphate (cAMP) and phorbol 12-myristate 13-acetate. The phenotypic effects of CRH in human skin cells are largely mediated by CRH-R1alpha through increases in concentrations of cAMP, inositol triphosphate (IP3), or Ca2+ with subsequent activation of protein kinases A (PKA) and C (PKC) dependent pathways. CRH also modulates the activity of nuclear factor of kappa light polypeptide gene enhancer in B-cells (NF-kappaB), activator protein 1 (AP-1) and cAMP responsive element binding protein (CREB). The cellular functions affected by CRH depend on cell type and nutritional status and include modulation of differentiation program(s), proliferation, viability and immune activity. The accumulated evidence indicates that cutaneous CRH is also a component of a local structure organized similarly to the hypothalamo-pituitary-adrenal axis. PMID- 16720313 TI - Gi proteins regulate lipopolysaccharide and Staphylococcus aureus induced cytokine production but not (1--> 3)-beta-D-glucan induced cytokine suppression. AB - Previous studies have demonstrated that bacterial lipopolysaccharide (LPS) and heat killed Staphylococcus aureus (SA) activation of inflammatory cells depended in part upon activation of heterotrimeric Gi proteins. It has also been shown that (1 --> 3) beta-D-glucan can suppress inflammatory cell activation by microbial products although the cellular mechanism of the glucan effect remains to be clearly defined. We hypothesized that Gi proteins function as a common convergent signaling pathway for both LPS and SA leading to monocyte mediator production. Additionally, we hypothesized that soluble glucan suppresses LPS and SA induced cytokine production via Gi protein coupled signaling. Human THP-1 promonocytic cells were pretreated with pertussis toxin (PTx, 100 ng/ml or 1 microgram/ml) 6 hours prior to stimulation with LPS (10 microgram/ml) and SA (10 microgram/ml) and/or soluble glucan (10 microgram/ml). Both LPS and SA significantly (p < 0.05) induced cytokine production IL-6 > TNF alpha > IL-1 beta > GM-CSF > IL-10 > IFN gamma. The induction of these cytokines was significantly (p < 0.05) suppressed by PTx. Glucan treatment alone had no effect on cytokine production but suppressed (P < 0.05) LPS and SA induced cytokines. PTx further augmented (p > 0.05) the inhibitory effect of glucan on the LPS and SA induced cytokine expression. The data support the hypothesis that Gi proteins function as a common signaling protein for both LPS and SA induction of pro-and anti inflammatory cytokines and that soluble glucan effectively suppresses cytokine production to the microbial stimuli. In contrast, the effect of soluble glucan on inhibiting cellular activation by LPS and SA is Gi protein independent. PMID- 16720314 TI - Growth inhibitory effects of quercetin on bladder cancer cell. AB - Quercetin, a flavonoid found in many fruits and vegetables, belongs to an extensive class of polyphenolic compounds. Previous studies reported that quercetin inhibits the proliferation of various cancer cells and tumor growth in animal models. We investigated the growth inhibition and colony formation of quercetin on three bladder cancer cells (EJ, J82 and T24). The expression of tumor suppressor genes and oncogenes such as P53, Survivin, PTEN, as well as the methylation status of these genes was also evaluated. We observed that quercetin induced apoptosis in bladder cancer cells in a time- and dose-dependent manner. Quercetin (100 micromolars) significantly inhibited EJ, T24 and J82 cell growth accompanied by an increase in the G0/G1 phase. In all cell lines, quercetin decreased the expression of mutant P53 and Survivin proteins. However, there was no change in the level of PTEN protein. Moreover, the DNA methylation levels of the estrogen receptor (Er-beta), P16INK4a and RASSF1A were strongly decreased (from 35 to 70%) in the quercetin-treated group compared to the control. In conclusion, our study suggested that quercetin inhibits growth, colony formation and hypermethylation of bladder cancer cell lines. Quercetin-induced apoptosis might be associated with a decrease in mutant P53 and Survivin proteins. PMID- 16720316 TI - Corticotropin-releasing hormone in nonhuman primates. AB - Understanding the many roles that corticotropin-releasing hormone (CRH) plays in facilitating the ordinary and extraordinary events that an individual faces during a lifetime is a complex task, and yet this knowledge is fundamental to understanding our own behaviour and physiology. During the past 25 years, the study of CRH in nonhuman primates, our closest genetic relatives, has grown rapidly. The intention of this review is to provide a broad overview of the research areas in which CRH has been investigated in monkeys and apes. The review begins with a detailed description of what we know about CRH, CRH receptors, and their distribution in the brain and periphery. The narrative then follows the life cycle, from the role of CRH in fertility, pregnancy and parturition, to the shaping of behaviour and neural processes by stressful experiences early in life. CRH is also examined in the context of its other regulatory roles, including appetitive behaviour and immune responses. Finally, the review examines the insights that nonhuman primate research offers us as to how CRH helps to shape our behaviour, whether it be our ability to socialize with our peers or to be a good parent. PMID- 16720317 TI - Angiogenic switch in Barrett's adenocarcinoma: the role of vascular endothelial growth factor. AB - The development of cancerous cells from the normal cells is the consequence of multiple genetic and epigenetic abuses. Activation of "Angiogenic switch" or formation of new blood vessels is one of the upshots of these abuses. Multiple factors are associated with the activation of angiogenic switch. Vascular endothelial growth factor (VEGF) and its down stream signaling molecules is important troupe of this event. In this article, we reviewed the role this troupe in the development of Barrett's adenocarcinoma and also discussed the possible remedies, which have the impact on blocking the function of this troupe. PMID- 16720318 TI - Phase I/II study of gemcitabine and epirubicine in stage IIIB-IV non small cell lung cancer. AB - Platinum-based chemotherapy currently represents standard treatment for advanced non-small cell lung cancer (NSCLC). Gemcitabine is one of the most promising agents currently in use in advanced NSCLC. As a single-agent, epirubicin, showed tumour response rates ranging from 17% to 36% in NSCLC. The aim of the present study was to evaluate the combination of gemcitabine and epirubicin in a phase I II study. Thirty chemotherapy-naive patients with stage III B-IV NSCLC received gemcitabine at a fixed dose of 1000 mg/m2 on days 1 and 8 every 3 weeks; epirubicin was administered every 21 days on day 1 at the initial dose of 80 mg/m2 which was subsequently escalated. Neutropenia was dose-limiting toxicity since it occurred in 3 out of five patients receiving epirubicin at the dose of 110 mg/m2. An objective response was observed in 14/30 patients, including 2 (7%) complete responses and 12 (40%) partial responses. Median duration of response was 12 months (range: 3 to 53 + months). Median overall survival was 16 months (range: 4 to 55 + months). The combination of gemcitabine and epirubicin is well tolerated. While the observed activity of this combinated treatment matches that of platinum-based regimens, the duration of response and survival have been sufficiently promising to initiate a phase II trial which is currently under way. PMID- 16720315 TI - Papillomavirus genome structure, expression, and post-transcriptional regulation. AB - Papillomaviruses are a group of small non-enveloped DNA tumor viruses whose infection usually causes benign epithelial lesions (warts). Certain types of HPVs, such as HPV-16, HPV-18, and HPV-31, have been recognized as causative agents of cervical cancer and anal cancer and their infections, which arise via sexual transmission, are associated with more than 95% of cervical cancer. Papillomaviruses infect keratinocytes in the basal layer of stratified squamous epithelia and replicate in the nucleus of infected keratinocytes in a differentiation-dependent manner. Viral gene expression in infected cells depends on cell differentiation and is tightly regulated at the transcriptional and post transcriptional levels. A noteworthy feature of all papillomavirus transcripts is that they are transcribed as a bicistronic or polycistronic form containing two or more ORFs and are polyadenylated at either an early or late poly(A) site. In the past ten years, remarkable progress has been made in understanding how this complex viral gene expression is regulated at the level of transcription (such as via DNA methylation) and particularly post-transcription (including RNA splicing, polyadenylation, and translation). Current knowledge of papillomavirus mRNA structure and RNA processing has provided some clues on how to control viral oncogene expression. However, we still have little knowledge about which mRNAs are used to translate each viral protein. Continuing research on post transcriptional regulation of papillomavirus infection will remain as a future focus to provide more insights into papillomavirus-host interactions, the virus life-cycle, and viral oncogenesis. PMID- 16720319 TI - Metabolism and ecology of purine alkaloids. AB - In this review, the biosynthesis, catabolism, ecological significance, and modes of action of purine alkaloids particularly, caffeine, theobromine and theophylline in plants are discussed. In the biosynthesis of caffeine, progress has been made in enzymology, the amino acid sequence of the enzymes, and in the genes encoding N-methyltransferases. In addition, caffeine-deficient plants have been produced. The ecology of purine alkaloids has not proved to be particularly promising. However, advances have been made in insecticidal and allelopathic fields, and in the role of microorganisms play in the changes that these compounds undergo in the soil. Caffeine inhibits cell plate formation during telophase throughout the development of coffee plants and other species. PMID- 16720320 TI - Delineation of the functional domains of the extracellular region of YWK-II Protein/APLP2 of sperm membrane. AB - The sperm membrane protein, designated as YWK-II protein/APLP2, is a member of the amyloid precursor protein (APP) superfamily and is a type I transmembrane protein involved in fertilization. Here, the structure-function of the domains of YWK-II protein was examined. Five segments with overlapping ends encompassing the entire extracellular region of mouse YWK-II gene were prepared, cloned and separately expressed in E. coli. The recombinant YWK-II segments were fused with glutathione S-transferase (GST), purified and evaluated for their antifertility activities by measuring their capacity to block in vitro mouse sperm-egg interaction. The structural domain(s) involved in the fertilization process was identified. The polypeptide segment corresponding to position 22-207 of YWK-II 763 inhibited the early stage of fertilization when the spermatozoa interacted with zona-free eggs; whereas the polypeptide segment 201-395 (lacking 309-364) of YWK-II-763 blocked sperm-egg membrane fusion. The remaining three segments, 201 395, 389-574 and 517-704 (lacking 613-624) of YWK-II-763, did not influence the in vitro fertilization process. The present results suggest that segment 22-308 of YWK-II-763 participates in the binding and fusion of sperm and egg plasma membranes thereby promoting fertilization. PMID- 16720321 TI - Protein C anticoagulant activity in relation to anti-inflammatory and anti apoptotic activities. AB - The anticoagulant protein C system is a dual function cofactor-dependent system. On one hand, it is designed to regulate coagulation, maintain the fluidity of the vasculature and prevent thrombosis. On the other hand, the protein C pathway provides anti-inflammatory and cytoprotective activities. Protein C, a vitamin K dependent serine protease zymogen that circulates in plasma, is converted by limited proteolysis to activated protein C (APC) by the thrombin-thrombomodulin endothelial protein C receptor complex on endothelial surfaces. APC and the cofactors of the protein C pathway exert two major distinct types of activities, namely a well-studied anticoagulant activity and a more recently revealed cytoprotective activity due to direct effects on cells. Because of these pleiotropic properties, APC and the protein C pathway components have important roles in the body's host-defense system and provide opportunities for therapeutic treatment of complex and challenging medical disorders, including thrombosis, severe sepsis and stroke. PMID- 16720322 TI - Cell adhesion molecules at the synapse. AB - Synapses are specialized intercellular junctions whose specificity and plasticity provide the structural and functional basis for the formation and maintenance of the complex neural network in the brain. The number, location, and type of synapses formed are well controlled, since synaptic circuits are formed in a highly reproducible way. This implies the existence of cellular and molecular properties that determine the connectivity of each neuron in the nervous system. Recent evidence has elucidated that these key features of the synapse are regulated by several families of cell-adhesion molecules (CAMs) enriched at synaptic junctions, including neuroligins, SynCAM, NCAM, L1-CAM, cadherins, protocadherins, and integrins. In this review we will discuss the various stages of synaptogenesis from the perspective of CAMs: Contact initiation, recruitment of presynaptic and postsynaptic proteins, synapse maturation/stabilization or elimination, and synaptic plasticity. We will also highlight some of the factors that regulate the function of these CAMs at the synapse, and discuss how dysfunction of these adhesive systems may contribute to several neurological disorders. PMID- 16720323 TI - Two dual specificity kinases are preferentially induced by wild-type rather than by oncogenic RAS-P21 in Xenopus oocytes. AB - In prior studies, we have found that oncogenic ras-p21 protein induces oocyte maturation using pathways that differ from those activated by insulin-induced wild-type ras-p21. Both oncogenic and wild-type ras-p21 require interactions with raf, but unlike oncogenic ras-p21, insulin-activated wild-type ras-p21 does not depend completely on activation of MEK and MAP kinase (MAPK or ERK) on the raf kinase pathway. To determine what raf-dependent but MAPK-independent pathway is activated by wild-type ras-p21, we have analyzed gene expression in oocytes induced to mature either with oncogenic ras-p21 or with insulin using a newly available Xenopus gene array. We find a number of proteins that are preferentially expressed in one or the other system. Of these, two proteins, both dual function kinases, T-Cell Origin Protein Kinase (TOPK) and the nuclear kinase, DYRK1A, are preferentially expressed in the insulin system. Confirming this finding, blots of lysates of oocytes, induced to mature with oncogenic ras p21 and insulin, with anti-TOPK and anti-DYRK1A show much higher protein expression in the lysates from the insulin-matured oocytes. Neither of these kinases activates or is activated by MAPK and is therefore an attractive candidate for being on a signal transduction pathway that is unique to insulin activated wild-type ras-p21-induced oocyte maturation. PMID- 16720324 TI - Epigallocatechin gallate, the main ingredient of green tea induces apoptosis in breast cancer cells. AB - Green tea has been suggested for prevention of cancers. In this study, the effect of the main constituent of green tea, epigallocatechin gallate (EGCG), on apoptosis of breast cancer cells was examined. EGCG induced apoptosis in T-47D cells through caspase cascade and the cells were detained at the G1 phase. The rate of apoptosis and activity of caspase-3 induced by EGCG was time and dose dependent. These findings suggest that EGCG might be useful in treatment and/or prevention of breast cancer by inducing apoptosis. PMID- 16720325 TI - Cellular immunology in HIV-1 positive African American women using alcohol and cocaine. AB - Co-use of illicit drugs, in particular cocaine and alcohol, is common among HIV 1(+) men and women of different ethnic groups. We compared cohorts of alcohol and cocaine co-users HIV-1(+) African American women and in cohorts of drug-free, or methamphetamine users HIV-1(+) men. We monitored clinical cellular immune parameters at repeated regular intervals. We found that significant inverse correlations between (CD8+)CD38+ cells and subpopulations of CD4+ cells distinguished by the expression of CD45RA in HIV-1(+) alcohol and cocaine co- users but not in drug-free HIV-1(+) patients. Following stratification for CD4+ cell number, we found the (CD4+)CD45RA+ subpopulation to be significantly higher (p < 0.05) in the drug user compared to drug-free HIV-1(+). Drug abuse may alter the change from the (CD4+)CD45RA+ to the (CD4+)CD45RA- phenotype selectively, which recovers in HIV-1+ methamphetamine abusers during treatment from baseline to 4-weeks, as manifested by improved IL-2 production in vitro. of TH1 and TH2 cytokines during progression to AIDS. PMID- 16720326 TI - WNT/Frizzled signaling in eye development and disease. AB - The canonical Wnt/Fzd signaling pathway is highly conserved among various species. Increasing evidence is accumulating for non-canonical Wnt signaling pathways, analogous to those discovered in Drosophila, being operative in vertebrates. Similarly, the networks of genes involved in eye development show significant conservation during evolution. The amenability of Drosophila for genetic manipulation and analysis of ocular phenotypes has delivered a great deal of information about the roles of the Wnt/Fzd signaling pathways at various stages of ocular development and growth, particularly in regulating the formation and size of the eye field, cell proliferation, polarity and differentiation. In addition to the numerous recent studies that have identified the expression of various components of these signaling pathways in the developing vertebrate eye, functional studies have revealed significant parallels in the way that Wnt/Fz signals regulate the formation of the vertebrate eye field and also the proliferation and differentiation of cells, particularly in the lens and retina. Significant advances have also recently been made in identifying mutations in these signaling pathways that underlie or contribute to various ocular diseases such as exudative vitreoretinopathy, retinal degenerations, cataract, ocular tumors and various congenital ocular malformations. Combined with the mechanistic studies in vertebrate and invertebrate models, these studies point to important functional roles for Wnt/Fzd pathways in the human eye. Further investigation of how these pathways function during eye development and growth may yield important insights into novel therapeutic approaches to treat or prevent diseases that cause blindness. PMID- 16720328 TI - Translesion synthesis DNA polymerases and control of genome stability. AB - Eukaryotic and prokaryotic genomes are replicated with amazingly high fidelity to assure faithful transmission of genetic information from one generation to the next. The accuracy of replication relies heavily on the ability of replicative DNA polymerases to efficiently select correct nucleotides for the polymerization reaction and excise mistakenly incorporated nucleotides using their intrinsic exonucleases. Cell also possess a variety of specialized DNA polymerases that help to overcome replication blocks when occasional unrepaired DNA lesions stall the replication machinery. The translesion synthesis (TLS) polymerases have an extremely low fidelity during copying undamaged DNA substrates, such that uncontrolled participation of these polymerases in DNA replication could present a threat to the genome stability. In this article, we discuss the properties of prokaryotic and eukaryotic TLS polymerases and their roles in modulating the rate of spontaneous and genotoxicant-induced mutations. We also review recent insights into the molecular mechanisms regulating the participation of error-prone TLS polymerases in the genome replication. Finally, we discuss the relationship between the functions of TLS polymerases and human disease. PMID- 16720327 TI - An oligonucleotide microarray study on gene expression profile in mouse testis of experimental cryptorchidism. AB - To investigate the germ cell apoptosis under body temperature in testis, we analyzed the gene expression patterns on day 1, day 4, day 7, day 14, day 28 and normal control adult mouse testis after experimental cryptorchidism (EC) using Affymetrix MOE430A microarray. Our data showed that EC led to the oxidative stress and gene expression fluctuation in the first 28 days, both of which were highly coincident in timing. Cryptorchid testis showed more effective antioxidative capability in the first 4 days, and suddenly lowered the capability from day 5 on, then gradually restored the antioxidation from day 10 to day 14, and turned to worse on day 28 again. The extensive high gene expression on day 4 after EC and the up-rising of oxidative stress level on day 5 and the abrupt down regulation of the gene on day 7 were closely related. From the chip data, we have found that the high level of reactive oxidative species (ROS) was not only related to the dysfunction or abnormality of the direct origin of ROS generation, but also related to the abnormality of the more upstream physiological events in energy metabolism, lipid metabolism. The selective regulation of metabolic substrate transporter in different cell population implied the existence of various regulation of the selective signal pathways among different cell populations by EC. PMID- 16720329 TI - Novel mutations of epidermal growth factor receptor in localized prostate cancer. AB - We recently demonstrated that EGFR protein overexpression is more common in African American (AA) prostate cancer patients compared to Caucasian patients. We further examine EGFR dysregulation by determining EGFR mutation status in the tyrosine kinase (TK) domain in prostate cancer patients of different ethnicity. Normal and tumor DNA from 89 radical prostatectomy cases were studied for mutations in the EGFR TK domain using genomic DNA sequencing. We identified 4 novel missense mutations in exons 19, 20 and 21 of EGFR TK domain: 3 in Koreans and 1 in Caucasian but none in AA. We also identified 5 distinct synonymous DNA sequence changes, which did not alter the encoded amino acid, in exons 20 and 21 in 31/89 (35%) patients. Interestingly, these synonymous sequence changes were not observed in normal DNA in 7(23%) patients, indicating the presence of de novo somatic mutation to a new synonymous sequence. Our data reveal that EGFR missense mutation in the TK domain occurs in localized prostate cancer. Our data also demonstrate the presence of somatic mutation to a new synonymous sequence in a subset of patients. Larger population-based studies are required to define the association between EGFR mutations and the ethnic background of patients. PMID- 16720331 TI - Corpora cavernosa as an alternative route for transfusion. AB - Routine intravenous blood transfusion is difficult when the blood pressure falls significantly or veins are inaccessible or are sclerotic due to multiple transfusions. Here, we describe the use of penile corpora cavernosa (CC), as an alternative route for blood transfusion and fluid replacement. The study was conducted in 15 men, 7 with massive burns, 6 with sclerotic veins from repeated injections, and 2 with extensive limb trauma. After the conventional methods of blood and fluid infusions were exhausted, a needle was inserted into CC for blood and fluid administration. During blood or saline infusions, penile shaft became elongated but returned to a normal length after termination of the infusion. There were no difficulties during needle insertion into CC, in varying the different transfusion rates, or in repetition of transfusion during the same or the subsequent days. Complications were rare with the exception of a subcutaneous penile hematoma in 2 patients which disappeared spontaneously. Erection was not disturbed in five patients who were followed for a mean of 10.4+/-1.8 months. These findings show that corpora cavernosa can be used for blood transfusion or for administration of fluids as a simple, easy, rapid, and safe vascular access in conditions in which conventional routes are inaccessible. PMID- 16720330 TI - Hydrogen peroxide acts as relaxing factor in human vascular smooth muscle cells independent of map-kinase and nitric oxide. AB - We previously showed that hydrogen peroxide (H2O2) induced resistance artery relaxation independent of endothelium. Thus, in this study we investigated the mechanism of relaxation induced by H2O2 on human renal vascular smooth muscle cell (HVSMC). HVSMC were stimulated with H2O2 and/or angiotensin II (Ang II), proline-rich-tyrosine-kinase-2 (PYK2), ERK1/2 MAP-Kinase, and myosin light chain 20 phosphorylation (Lc20) were assessed using Western blot analysis in the presence of potassium channel blockers, MAP-Kinase, and nitric oxide synthesis (NOS) inhibitors. H2O2 increased PYK2 and ERK1/2 phosphorylation, and at the same time decreased Lc20 phosphorylation. AngII increased phosphorylation of PYK2, ERK1/2 and Lc20, whereas, the pretreatment of HVSMC with H2O2 decreased Lc20 phosphorylation induced by AngII. MEK inhibition, decreased ERK1/2 phosphorylation, but had no effect on the inhibition of phosphorylation of Lc20 induced by H2O2. The inhibition of Ca2(+)-dependent K+ channels (BKCa) and NOS did not block the decrease of Lc20 phosphorylation in response to H2O2. On the other hand, pretreatment of HVSMC with 60 mM of KCl, increased rather than decreased Lc20 phosphorylation in response to H2O2. This study shows the evidence that H2O2 acts as a relaxing factor and as an activator of PYK2 and ERK1/2 in Human renal VSMC. The relaxation induced by H2O2 is independent of BKCa, ERK1/2 MAP-Kinase and NOS pathways. The relaxing effect to H2O2 changes to contracting effect when the potassium channels are compromised. PMID- 16720332 TI - Proteolytic enzymes in skeletal development: histochemical methods adapted to the study of matrix lysis during the transformation of a "cartilage model" into bone. AB - The replacement of a "cartilage model" by definitive bone is characterized by a series of localized excavations of the cartilage which are eventually followed by bone deposition. Each excavation requires lysis of cartilage components (defined here as the breakdown of a peptide bond) and their eventual resorption (defined here as microscopical visible cartilage loss). More precisely we have proposed that the lysis is affected by proteases capable of breaking down the main proteoglycan "aggrecan" and the main fibril element, "type II collagen". Four approaches combining biochemical, immunologic and microscopic techniques have been adapted to test this hypothesis. Each is applied to the rat tibial head's "cartilage model" where proteases have been shown to be major contributors to secondary ossification center formation. The approaches have been found both effective and distinct as cartilage resorbing enzymes have not only been identified but also detected in situ before and after activation. Achieved overall is an understanding of when, where and how specified proteases contribute to tissue component lyses. While the focus resides on the in situ proteolysis of cartilage, three of the approaches could be translated without change to other tissues, whereas one may require tissue specific adjustments before use. PMID- 16720333 TI - Mitochondrial Ca2+ transport, permeability transition and oxidative stress in cell death: implications in cardiotoxicity, neurodegeneration and dyslipidemias. AB - Mitochondrial Ca2+ transport is important in the maintenance of intracellular ion homeostasis, and also a key factor in the pathogenesis of many diseases. We discuss here the main aspects of mitochondrial Ca2+ transport, and how this transport is linked to changes in energy metabolism and redox state. Mitochondrial permeability transition, a consequence of excessive mitochondrial Ca2+ accumulation associated with oxidative stress is also discussed. Finally, our current understanding of the involvement of these mitochondrial processes in cardiac ischemia-reperfusion, neurodegeneration and dyslipidemias is presented. PMID- 16720334 TI - Up date of in vitro production of porcine embryos. AB - There have been intensive attempts to establish reliable in vitro maturation (IVM) and fertilization (IVF) methods in pigs. Although a great deal of progress has been made, current IVM-IVF systems still suffer from a low rate and poor quality of in vitro produced embryos . In this review, we will review the recent studies about IVM-IVF of porcine oocytes and the in vitro culture (IVC) system, especially modified in vitro production (IVP) system that produces high quality of porcine blastocysts. We then try to suggest practical ways to solve the problems mentioned above in the pigs. PMID- 16720335 TI - Nutritional and medicinal use of Cactus pear (Opuntia spp.) cladodes and fruits. AB - Natural products and health foods have recently received a lot of attention both by health professionals and the common population for improving overall well being, as well as in the prevention of diseases including cancer. In this line, all types of fruits and vegetables have been re-evaluated and recognized as valuable sources of nutraceuticals. The great number of potentially active nutrients and their multifunctional properties make cactus pear (Opuntia spp.) fruits and cladodes perfect candidates for the production of health-promoting food and food supplements. Although traditionally appreciated for its pharmacological properties by the Native Americans, cactus pear is still hardly recognized because of insufficient scientific information. However, recent studies on Opuntia spp. have demonstrated cactus pear fruit and vegetative cladodes to be excellent candidates for the development of healthy food. Therefore, this review summarizes current knowledge on the chemical composition of Opuntia cacti with particular emphasis in its use as food and medicine. PMID- 16720337 TI - Mechanisms controlling CDK9 activity. AB - This review primarily focuses on the mechanisms that modulate CDK9 activity and its recruitment to cellular genes, where it phosphorylates the C-terminal domain of RNA polymerase II (RNAPII) as well as negative elongation factors. CDK9 associates with each of four cyclins (T1, T2a, T2b and K), forming distinct positive transcription elongation factors (P-TEFb). Research done during the past decade has demonstrated a role for P-TEFb in stimulating elongation of otherwise paused RNAPII transcripts. Recent work suggests that P-TEFb also positively modulates other steps during transcription. In addition, "abnormal" CDK9 function is associated with certain diseases. Specifically, the activity of the cyclin T1/CDK9 complex is essential for HIV-1 replication and CDK9 upregulation is associated with cardiac hypertrophy. Thus, the role of CDK9 in these processes, and the possibility of therapeutically targeting CDK9, will also be briefly discussed. PMID- 16720336 TI - Smoking cessation program and CYP2A6 polymorphism. AB - The relationship between CYP2A6 genotype and smoking status remains unclear although several studies have been reported. In this study, we have investigated the significance of CYP2A6 genotype on smoking habit and treatment of nicotine patch. Sixty-one smokers (1.7%) working in a Japanese company (n = 3585) participated in this smoking cessation program. Forty-four of 61 (72.1%) smokers were treated by nicotine patch. A genotyping analysis was carried out for 41 (40 men and 1 women) of 61 participants (67.2%). The smoking cessation rate at 90 days was 54.1% (33/61). Age and smoking years in re-smoking group are significantly lower than those in smoking cessation group. The smoking cessation rate of participants treated with nicotine patch (63.6%; 28/44) was significantly higher than that of the group non-treated with nicotine patch (29.4%; 5/17) at 90 days (p < 0.05). The incidence of homozygotes of CYP2A6 gene deletion (CYP2A619934/19934) in 41 cases (9.8%; 4/41) could be higher than that in 894 healthy controls (3.7%; 33/894) (p = 0.12), while no other variant alleles (CYP2A619932, CYP2A619933 and CYP2A619936) were found. Age and smoking years of participants with CYP2A619934/19934 are significantly higher than those with CYP2A619931 positive. The scores of Fagerstrom test, an analysis for nicotine dependence, were slightly different between participants with CYP2A619934/19934 and CYP2A619931 positive. Although treatment of nicotine patch is efficacious to smoking cessation, cases with CYP2A619934/19934 might be more sensitive to nicotine adverse effects and more difficult to quit smoking once they have smoking habit. PMID- 16720338 TI - In vitro propagation of spermatogonial stem cells from KM mice. AB - Spermatogonial stem cells (SSCs) are a unique type of stem cells in that they transmit genetic information to the next generation by producing sperms. Studies of SSC proliferation and differentiation have been hampered by the inability of reconstructing these processes in vitro, particularly in a serum-free culture system. Several groups have reported the long term culture of SSCs during which SSCs self-renew and restore spermatogenesis when transplanted back to recipient testes. However, different protocols and mice with particular genetic background have been used by different laboratories, and the techniques have not been adopted widely. In the present study, we first established a SSC isolation and culture system composed of differential adherence selection of SSCs, serum-free medium and mouse embryonic fibroblast (MEF) feeder cells. SSCs from KM pups could be cultured on MEF feeders in StemPro-34 SFM Medium supplemented with glial cell line-derived neurotrophic factor (GDNF), soluble GDNF family receptor alpha-1 (GFRa1) and basic fibroblast growth factor (bFGF) for 1 month. These SSCs were characterized morphologically and by examining the expression of marker genes. Expression of Oct4 and Sox2, which are crucial factors in embryonic stem cell (ESC) self-renewal, were detected in our cultured SSCs, suggesting that SSCs may share with ESCs some common mechanisms in self-renewal regulation. We also found that LIF had no effect on the proliferation of cultured SSCs derived from KM mice. PMID- 16720339 TI - Mutation of siRNA results in thermodynamically unstable duplex which influences knockdown of dmrt1 by RNA interference. AB - MicroRNAs (miRNAs) constitute a growing class of non-coding RNAs that are thought to regulate gene expression by translational repression and mRNA degradation. We report here that short interfering RNA (siRNA) mutation significantly changed kinetics of the folding and unfolding of secondary structures and decreased Tm value of the duplex melting. The mutant duplex was more unstable thermodynamically than the normal structure. Furthermore negative effects of the mutation on RNA interference (RNAi) was observed in both mouse dmrt1 transfected COS-7 and Sertoli cells in which endogenous dmrt1 was expressed. However, interference efficiency of mutational and normal duplex in Sertoli cells was not significant in comparison with those in dmrt1 transfected COS-7 cells, suggesting complex regulatory mechanisms in RNAi in endogenous dmrt1 expression. Abundance of intronic miRNA structures observed in the dmrt1 gene may also contribute to the precise regulation cascade in the dmrt1 expression. These findings help in further understanding RNA silence in vertebrate development and drug design for target gene. PMID- 16720340 TI - Identification and analysis of novel genes expressed in the mouse embryonic facial primordia. AB - Craniofacial anomalies are a common feature of human congenital dysmorphology syndromes, suggesting that genes expressed in the developing face are likely to play a wider role in embryonic development. To facilitate the identification of genes involved in embryogenesis, we previously constructed an enriched cDNA library by subtracting adult mouse liver cDNA from that of embryonic day (E)10.5 mouse pharyngeal arch cDNA. From this library, 273 unique clones were sequenced and known proteins binned into functional categories in order to assess enrichment of the library (1). We have now selected 31 novel and poorly characterised genes from this library and present bioinformatic analysis to predict proteins encoded by these genes, and to detect evolutionary conservation. Of these genes 61% (19/31) showed restricted expression in the developing embryo, and a subset of these was chosen for further in silico characterisation as well as experimental determination of subcellular localisation based on transient transfection of predicted full-length coding sequences into mammalian cell lines. Where a human orthologue of these genes was detected, chromosomal localisation was determined relative to known loci for human congenital disease. PMID- 16720341 TI - Plasma epigenetic markers for cancer detection and prenatal diagnosis. AB - Successful detection of circulating nucleic acids has opened up new possibilities in cancer testing and prenatal diagnosis. Circulating DNA markers are useful in cancer detection, prognostication and monitoring. Cancer-associated molecular changes which can be detected include gene mutations, chromosomal rearrangements, microsatellite alterations, viral sequences, and, to be discussed in more detailed, gene promoter hypermethylation. Methylation analysis is commonly performed by DNA digestion with methylation-sensitive restriction endonucleases followed by polymerase chain reaction (PCR), or bisulfite modification followed by methylation-specific PCR (MSP). The detection of fetal DNA in maternal plasma has opened up new possibilities for noninvasive prenatal diagnosis. However, circulating fetal DNA detection has been based on exploiting gender and polymorphic differences between the fetus and mother. The recent discovery of epigenetic differences between the maternal and the fetal DNA detectable in maternal plasma has launched a hunt for fetal-derived epigenetic markers in maternal plasma. It is hoped that this type of universally applicable markers would be made available in a clinical diagnostic setting in the near future. PMID- 16720342 TI - Werner syndrome: molecular insights into the relationships between defective DNA metabolism, genomic instability, cancer and aging. AB - Werner syndrome is a segmental progeroid disease characterized by increased cancer and acceleration of specific age-related phenotypes, due to loss of a protein known as WRN. Extensive research over the last decade has revealed much about WRN biochemistry and the etiology of Werner syndrome. WRN possesses multiple DNA-dependent enzymatic activities (ATPase, helicase, exonuclease, and strand annealing) and interacts with factors having established roles in DNA metabolic pathways. Although the exact functions of WRN remain unclear, accumulating evidence points to roles in proper resolution of replication blockage and in telomere maintenance. If WRN function is lost (as exemplified in cells from Werner patients), problems with replication and DNA damage processing arise, probably resulting in an increased number or persistence of strand breaks. In turn, these events lead to chromosomal and telomeric abnormalities or activate checkpoints that bring about early senescence or increased apoptosis. Thus, elevated cancer incidence associated with Werner syndrome is due to increased chromosomal changes, while the accelerated aging characteristics probably stem from telomere dysfunction leading to accumulation of non-functional senescent cells or excessive apoptotic cell death over time. More research is needed to determine whether these specific DNA-dependent mechanisms contribute to development of aging characteristics in normal individuals. PMID- 16720343 TI - Epstein-Barr virus (EBV) and its associated human cancers--genetics, epigenetics, pathobiology and novel therapeutics. AB - Epstein-Barr virus (EBV) is a B-lymphotropic virus that is associated with a range of human malignancies. Although for many of these tumors the association has long been established, unraveling the precise role of EBV in disease pathogenesis has been more difficult. This review summarizes current knowledge concerning the association between EBV and human cancers, and illustrates how a deeper insight into viral latent gene expression, regulation and functions in different cell environments is already helping towards a better understanding of both the natural history of infection in normal individuals and how EBV contributes to malignant transformation. Finally, therapeutic strategies targeting EBV in tumors are discussed. PMID- 16720344 TI - Dual specificity phosphotase 18, interacting with SAPK, dephosphorylates SAPK and inhibits SAPK/JNK signal pathway in vivo. AB - The SAPK/JNKs play important roles in numerous cellular processes, and for this reason they have become putative drug targets. Most dual-specificity protein phosphatases (DSPs) play important roles in the regulation of mitogenic signal transduction and cell cycle control in response to extracellular stimuli. Dual specificity phosphatase 18 (DUSP18), a newly recognized SAPK/JNK phosphatase, is widely expressed. This expression is modulated in response to extracellular stimuli. By phosphorylation assay, pull down and coimmunoprecipitation experiments, it is shown here that DUSP18 interacts with SAPK/JNK and dephosphorylates it both in vitro and in vivo. DUSP18 does not dephosphorylate p38 or p44ERK1. Furthermore, DUSP18 inhibits SAPK/JNK pathway in vivo. Based on these findings, DUSP18 appears to serve an important role by regulation of SAPK/JNK pathway. PMID- 16720345 TI - Critical overview of mitochondrial nitric-oxide synthase. AB - The recent discovery of mitochondrial nitric-oxide synthase (mtNOS) has provided novel information on mitochondrial biology. mtNOS, localized at the inner membrane, functions as one of the important regulatory factors that affect cellular respiration in mitochondria. The distribution of mtNOS in diverse organs of many species suggests its biological importance. Changes in mtNOS expression and activity in many pathophysiological situations may imply its significant involvement in various NO-related biological phenomena. The functional coordination of mtNOS with other NOSs present in the same or adjacent tissues is unknown as it is most of the regulatory mechanisms of mtNOS expression. Thus, future studies will be required to validate the physiological significance of mtNOS, and to elucidate its regulatory mechanisms on cellular energy metabolism in mitochondria. PMID- 16720346 TI - Atheroma development in apolipoprotein E-null mice is not affected by partial inactivation of PTEN. AB - PTEN is a dual-specificity phosphatase that has been shown to inhibit vascular smooth muscle cell (VSMC) proliferation and migration, two key events in the ethiopathogenesis of atherosclerosis. Adenovirus-mediated PTEN overexpression inhibited the formation of vascular obstructive lesions induced by mechanical injury of the vessel wall. In this study, we investigated whether PTEN protects against atheroma formation in apolipoprotein E-null mice (apoE-/-), a widely used animal model characterized by the development of hypercholesterolemia and atherosclerosis. We examined atheroma development in the aorta of apoE-/- mice with an intact Pten gene and apoE-/- mice lacking one allele of Pten (Pten(+/ )apoE-/-) that were challenged for six weeks with an atherogenic diet. Compared with apoE-/- controls, Western blot analysis of arterial cell lysates from Pten(+/-)apoE-/- mice revealed a decrease in PTEN expression. This correlated with increased phosphorylation of AKT, thus demonstrating that Pten inactivation in Pten(+/-)apoE-/- mice has functional consequences. However, the extent of atherosclerosis was undistinguishable in both groups of fat-fed mice. Likewise, the atheroma of Pten(+/-)apoE-/- and apoE-/- mice displayed similar VSMC content, cellularity and rates of proliferation and apoptosis. Thus, in spite of the cytostatic and antimigratory activities of PTEN, and in contrast to previous studies demonstrating that Pten is haplo-insufficient for tumor suppression, our results demonstrate that atherosclerosis in hypercholesterolemic mice is not aggravated by partial inactivation of Pten. PMID- 16720347 TI - Remodeling of neuronal networks by stress. AB - Stress can be a threat to the physiological and psychological integrity of an individual and may result in psychic and behavioral changes. The stress response is mediated through in-concert activity of many brain areas and there is experimental evidence that stress induces structural changes in neuronal networks, in particular in the hippocampus, the prefrontal cortex and the amygdala. Within the hippocampal formation, stress exposure results in remodeling of dendrites of the CA3 pyramidal neurons and in reduced numbers of synapses on these neurons. Furthermore, stress inhibits adult neurogenesis in the dentate gyrus and appears to modulate the GABAergic system. In the prefrontal cortex, repeated exposure to stress causes dendritic retraction and loss of spines in pyramidal neurons whereas in the amygdala stress can elicit dendritic hypertrophy. These microscopically detectable changes in neuronal structures indicate the reorganization of neuronal networks. Moreover, molecular studies show that stress modulates expression of genes involved in neuronal differentiation and/or structural remodeling. Since a wealth of data documents the adverse effects of stress on emotions and cognition these alterations are commonly interpreted as the deleterious effect of chronic stress on the central nervous system. However, it is also possible that at least part of these changes reflect adaptive responses, as the network system rearranges its connections in order to cope with the changing requirements from the internal or external environment. PMID- 16720349 TI - Role of aldo-keto reductases in development of prostate and breast cancer. AB - Aldo-keto reductase (AKR) is a super gene family, consisting of fourteen families and more than 40 members overall. These proteins have been well known as metabolic enzymes of carbonyls, but recent data indicates that the members in AKR families 1 and 7 (AKR1 and AKR7) are involved in the development of some human and rodent tumors, such as in primary liver, lung, colorectal, prostate, and breast cancers. They are involved in the pathogenesis, diagnosis, and therapy of these tumors. This manuscript discusses the recent progression in AKR study in mammalian tumors, focusing on prostate and breast cancer. PMID- 16720350 TI - Changes in gene expression with increased transglutaminase 2 in a SH-SY5Y cell line. AB - Previously we showed that the overexpression of transglutaminase 2 (TGase 2) resulted in activation of NF-kappaB through polymerization of I-kappaBalpha. To explore the pathway of TGase 2-mediated NF-kappaB activation, a transcriptomic microarray analysis was performed. In a SH-SY5Y cell line transfected with TGase 2, 24 genes were up-regulated at least 1.6-fold and 26 genes were down-regulated, as compared to the wild-type cell line. Detailed analysis resulted in the identification of target genes involved in regulating inflammation, including tribbles homolog 3, peroxiredoxin 4, neuropeptide Y, galanin, and vasoactive intestinal peptide. Our data demonstrate that the increase in TGase 2 in the neuroblastoma causes functional changes in transcriptional regulation, especially in genes associated with inflammation. These changes in gene expression caused by increases in TGase 2 activity may contribute to the pathophysiologic processes of inflammatory diseases. PMID- 16720348 TI - Uridine phosphorylase in breast cancer: a new prognostic factor? AB - Uridine phosphorylase (UPase) is an enzyme that converts the pyrimidine nucleoside uridine into uracil. Upon availability of ribose-1-phosphate, UPase can also catalyze the formation of nucleosides from uracil as well as from 5 fluorouracil, therefore involved in fluoropyrimidine metabolism. UPase gene expression is strictly controlled at the promoter level by oncogenes, tumor suppressor genes, and cytokines. UPase activity is usually elevated in various tumor tissues, including breast cancer, compared to matched normal tissues and this induction appears to contribute to the therapeutic efficacy of fluoropyrimidines in cancer patients. In this review, we will discuss in detail the role of UPase in the activation of fluoropyrimidines and its effect on the prognosis of breast cancer patients. PMID- 16720351 TI - Simultaneous in vivo measurement of dopamine, serotonin and ascorbate in the striatum of experimental rats using voltammetric microprobe. AB - The in vivo performance of a voltammetric microprobe based on overoxidized poly(1,2-phenylenediamine) coated carbon fiber microelectrode (OPPD/CFME), developed in our laboratory, is presented. For this purpose, an OPPD microprobe was stereotaxicaly implanted in the striatum of a deeply anesthetized Wistar rat for the simultaneous measurement of dopamine, serotonin and ascorbate. Furthermore, the post mortem levels of these physiologically important compounds were monitored after the rats were terminated with an overdose of anesthetic introduced through an indwelling jugular catheter. Using cyclic (CV) and square wave (SWV) voltammetry, the OPPD/CFME was demonstrated to exhibit efficient separation of the voltammetric signals of dopamine, serotonin and ascorbate in the presence of biological matrix, with the SWV mode allowing more convenient detection regarding both sensitivity and selectivity. Explicit proof of in vivo dopamine detection at the OPPD/CFME was achieved via the absence of the dopamine signal in rats with unilateral lesions of nigrostriatal dopaminergic neurons, which was induced by the use of the selective dopaminergic neurotoxin 6 hydroxydopamine (6-OHDA). In stark contrast to non-treated rats, where a strong signal corresponding to 1.2 micro M dopamine was measured at ca. 90 s after the rats' death, the signal for dopamine in dopamine-depleted striatum of 6-OHDA rats was absent. A critical comparison of the in vivo performance of the OPPD/CFME to that of bare and Nafion-coated carbon fiber microelectrodes, often used in neurophysiological studies, clearly showed a significant advantage of the former microprobe. The OPPD/CFME allowed multiple and repetitive in vivo measurements, along with pre- and post-measurement external calibration, with no loss in selectivity and an acceptable loss in sensitivity, indicating that the active sensing sites were not adversely blocked by the components of the extracellular fluid, thus affirming the great practical in vivo applicability of the OPPD microprobe. PMID- 16720352 TI - Demonstration of a physiologic sphincter at duodeno-jejunal junction. AB - Current evidence suggests that there are three sphincters that regulate the flow of chyme from stomach to the duodenum. We investigated the hypothesis whether a fourth 'physiologic sphincter' exists at the duodeno-jejunal junction. The pressure response of the jejunum, duodeno-jejunal junction and duodenum to individual balloon distension of the jejunum and duodenum was recorded in 28 healthy subjected before and after anesthesizing the jejunum, duodeno-jejunal junction and duodenum. The duodeno-jejunal junction length was measured by the station pull-through technique. Duodenal balloon distension with 2 and 4 ml of normal saline did not change the pressures in the duodenum, duodeno-jejunal junction or jejunum (p > 0.05). Distension with 6 ml saline produced an increase of duodenum pressure (p < 0.01), a decrease of duodeno-jejunal junction pressure (p < 0.01), and no change in the pressure of the jejunum (p > 0.05), the balloon was expelled to the jejunum. Eight, and 10 ml duodenum balloon distension produced pressure changes similar to those of the 6 ml distension (p > 0.05). Jejunum balloon distension with 2 and 4 ml saline induced no jejunum, duodeno jejunal junction or duodenum pressure changes (p > 0.05). Six ml balloon distension effected increase of jejunum (p < 0.01) and duodeno-jejunal junction (p < 0.05) pressure, but no duodenum pressure changes (p > 0.05). Jejunum balloon distension with volumes more than 6 ml produced pressure changes similar to the 6 ml distension. Distension of the anesthetized duodenum, duodeno-jejunal junction or jejunum did not change the duodeno-jejunal junction pressure. A high pressure zone of 1.6+/-0.04 cm length was detected at the duodeno-jejunal junction. Together, the findings show that a high pressure zone exists at the duodeno jejunal junction suggesting that this region might act as a physiological sphincter. PMID- 16720353 TI - pH sensitive photophysical and photochemical properties of a pentaazadentate porphyrin-like gadolinium (III) complex. AB - The electronic absorption spectra, photobleaching property, emission, and triplet excited state characteristics of a pentaazadentate porphyrin-like gadolinium(III) complex have been investigated at acidic and basic conditions. The electronic absorption spectrum of this complex exhibits a Soret band at ca. 449 nm and a Q like band at ca. 880 nm when the pH value of the solution is lower than 6.6. At basic conditions, the Q-like band blue shifts to ca. 624 nm, while the Soret band only shows an approximate 20 nm hyposochromic shift. The acidic solution is relatively stable upon exposure to ambient light, but the basic solution photobleached to colorless in approximately 3 hours. Irradiation of the Soret band of basic solutions at 420 nm causes faster photobleaching than irradiation of the Q-like band (624 nm). The emission of this complex at pH = 6.5 appears at ca. 915 nm (max.) and 1016 nm, which is a mirror image of the Q-like band, indicating the nature of the emitting state being the lowest singlet excited state. At pH = 9.0, the emission band shifts to ca. 585 nm (max.) and 630 nm (shoulder). The triplet transient difference absorption spectrum of the solution at pH = 6.5 exhibits a bleaching band at ca. 460 nm, a narrow positive band at ca. 450 nm, and a broad, moderately intense absorption band from 480 nm extending to the near-IR region (700 nm). The triplet excited state lifetime deduced from the decay of the transient absorption is approximately 81 ns. PMID- 16720354 TI - Response and function of skeletal muscle heat shock protein 70. AB - In response to stress, cells produce a series of heat shock proteins (Hsps). One of the most prominent Hsps, is the 70 kDa Hsp (Hsp70). Hsp70 is a highly conserved and essential protein against stress. The skeletal muscle responds to a diverse group of stress signals namely, muscle contraction linked energy and milieu challenges, ischemia and exercise by producing Hsp70. The extent of this Hsp70 response in skeletal muscle depends on the type and intensity of the signal, and is characterized in a muscle fiber specific manner by a special time course. Hsp70 in the skeletal muscle is regulated at transcriptional, translational and posttranslational levels. Hsp70 serves as an indicator for cellular stress as a molecular chaperone, plays pivotal role in maintaining cellular homeostasis by preventing apoptosis, influences energy metabolism, facilitates cellular processes in terms of muscular adaptation and interacts with other signalling pathways. This review summarizes our current knowledge on the skeletal muscle Hsp70 response. PMID- 16720355 TI - Animal models for perinatal transmission of HIV-1. AB - Despite progress in the use of antiretroviral drugs, mother-to-child transmission of HIV still remains a serious medical problem in resource-poor areas. There is a need to find the best method for drug delivery to reduce transmission, while keeping the risk of selection for drug-resistant viral variants low. Even when infection is prevented during pregnancy, the risk of acquiring infection by breast feeding remains significant and in some settings, is unavoidable. The ability of antiretroviral drugs or vaccines to limit transmission by breast milk is unknown. HIV vaccines are still in an early phase of development and have not yet been tested in newborns, in part due to concerns about potential of low immunogenicity due to transplacental transfer of maternal antibodies. Alternative strategies have been proposed to limit transmission using passive prophylaxis by human monoclonal antibody, but to insure product safety, trials have been slowed. Due to such concerns, animal models may provide an alternative for testing efficacy in human newborns. In this review, advances made using such models will be compared for mother-to-child transmission of lentivirus with that of HIV-1. In addition, some perspectives on integrating the data obtained from these models as a groundwork for future clinical work will be presented. PMID- 16720356 TI - The role of tumor metastasis suppressors in cancers of breast and prostate. AB - Despite significant improvement in surgical techniques and chemotherapies, none of the current medical technologies "cure" metastatic disease, and the patients who have acquired metastatic cancer inevitably die from disseminated disease. Thus, there is a need for developing novel therapeutic approaches which can directly target metastatic tumor cells. However, advances in understanding the molecular mechanism of tumor metastases have lagged behind other developments in the cancer field. Tumor metastasis involves complex array of steps with each step requiring a coordination of the actions of many positive and negative factors. A number of tumor metastasis suppressors have been identified which suppress the formation of tumor metastasis without affecting the growth rate of the primary tumor. Such discoveries offer new approaches for curtailing tumor metastasis. This review summarizes our current understanding on these genes and their potential role in the progression of tumor metastases. PMID- 16720357 TI - Separate and combined effects of Cu and Cd on seedling growth and active oxygen metabolism system of Trifolium repens L. AB - Pot-culture experiments were used to examine the individual and combined effects of Cu and Cd pollutants on Trifolium repens L. seedlings, both on their growth and their active oxygen metabolism system, mainly superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) activities. The results showed that the negative action took place at low concentrations of Cu (less than 500 ppm) and Cd (less than 0.5 ppm), which had no obvious effects on the seedlings' growth. However, as the concentrations of Cu and Cd increased (500-3000 ppm and 0.5-50 ppm respectively), synergistic activities was observed, showing obvious negative effects (P less than 0.05). Compared with the control samples, the seedlings affected by Cu and Cd pollutants were shorter and smaller, their fresh/dry weight and content of soluble protein decreased drastically, their leaf electric conductivity increased, and the contents of their leaf pigments decreased. Chlorophyll a was more sensitive than chlorophyll b to Cu and Cd pollutants, and chlorophyll b was more sensitive than carotenoid. It was also shown that the active oxygen metabolism of T. repens seedlings was destroyed by high amounts of Cu and Cd, the balance of the anti-oxidase system was broken, and the CAT and SOD activities noticeably decreased while POD activity evidently increased. Cd had a more noticeable effect on seedling growth than Cu. PMID- 16720358 TI - Effects of IL-1 beta on RT1-A/RT1-DM at the maternal-fetal interface during pregnancy in rats. AB - Interleukin-1 (IL-1beta) beta and major histocompatibility complex (MHC) play an important role during pregnancy. Expression of non-classical class MHC II RT1-DM antigen and classical class MHC I RT1-A antigen induced by IL-1beta was examined by Northern blotting, Western blotting and immunohistochemistry. IL-1beta treatment significantly increased the expression of RT1-A and RT1-DM in early and mid pregnancy. In late pregnancy, expression of RT1-DM significantly increased in uteri and decreased in placenta. Immunohistochemical studies indicated that, in early pregnancy, RT1-DM protein mainly localized to luminal and glandular uterine epithelium, and RT1-A was present in deciduas basalis, outer layer of luminal epithelium and glandular epithelium. During mid and late pregnancy, RT1-DM was present in maternal blood vessels and syncytiotrophoblast of labyrinthine zone, and RT1-A was present in maternal blood vessels and trophoblastic epithelium of the labyrinthine layers. These findings show that exogenous IL-1beta affects expression of RT1-DM and RT1-A and does not affect the localization of corresponding molecules during pregnancy. PMID- 16720359 TI - Cyclooxygenase 2: understanding the pathophysiological role through genetically altered mouse models. AB - Cyclooxygenase (COX) -1 and -2 catalyze the first step in the biosynthesis of prostanoids. COX-1 is constitutively expressed in many tissues and seems to be involved in the housekeeping function of prostanoids. COX-2, the inducible isoform, accounts for the elevated production of prostaglandins in response to various inflammatory stimuli, hormones and growth factors. COX-2 expression has been also associated with cell growth regulation, tissue remodelling and carcinogenesis. More of these characteristics have been elucidate through using COX selective inhibitors. Recent advances in transgenic and gene-targeting approaches allow a sophisticated manipulation of the mouse genome by gene addition, gene deletion or gene modifications. The development of COX-2 genetically altered mice has provided models to elucidate the physiological and pathophysiological roles of this enzyme. PMID- 16720360 TI - The genetic and environmental factors involved in benzidine metabolism and bladder carcinogenesis in exposed workers. AB - Genetic susceptibility to bladder cancer in individuals exposed to arylamines may be explained by interindividual metabolic differences that lead to arylamine bioactivation or detoxification. In this article, occupational bladder cancer risk factors and the evidence that links benzidine exposure to bladder cancer are reviewed. Benzidine metabolism is described and compared with that of other aromatic amines. Metabolic polymorphisms and bladder cancer in the context of occupational exposure to aromatic amines are also reviewed, and the environmental and genetic relationships of benzidine exposure and genetic susceptibility are outlined. Only a few studies of bladder cancer genetic susceptibility in populations exposed occupationally to arylamines have been published. The results of these case-control studies show conflicting results, reflecting metabolic differences between monoarylamines and diarylamines such as benzidine. Additional studies and pooled analyses of existing data are needed to establish if individuals are at higher risk of bladder cancer given the presence of certain alleles that make them more susceptible to this disease. PMID- 16720361 TI - MUC1 is a scaffold for selectin ligands in the human uterus. AB - MUC1 is a large, transmembrane mucin glycoprotein abundantly expressed at the apical surface of uterine epithelia in all species examined to date. Loss of MUC1 at the time of embryo implantation occurs in many species; however, this does not appear to be the case in humans. Recent studies indicate that human blastocysts express L-selectin at their external surfaces raising the possibility that selectin ligands expressed at the apical surface of the uterine epithelium support early stages of blastocyst attachment. In the current study, we have used a panel of antibodies specific for selectin ligands to determine if MUC1 functions as a scaffold for these carbohydrate motifs in fertile women. The results demonstrate that MUC1 carries selectin ligands throughout the secretory phase of the menstrual cycle, including the mid-secretory (receptive) phase. Consequently, MUC1 represents a potential ligand for selectins expressed by human blastocysts. PMID- 16720362 TI - Steroid hormone mediated regulation of corticotropin-releasing hormone gene expression. AB - Corticotropin-releasing hormone (CRH), a 41 amino acid polypeptide, is expressed in many regions of central nervous system and peripheral tissues and mediates many physiological functions. Abnormal production of CRH is involved in some pathological processes. Among various endogenous factors that modulate CRH production, steroid hormones control CRH production by regulating its gene transcription. Although there is no classical steroid hormone response element in the CRH promoter, steroid hormones regulate CRH gene expression through protein protein interaction or by binding directly to response elements. PMID- 16720363 TI - Clinical trials. AB - A clinical trial is a research study in human patients aimed at answering specific health questions. If a clinical trial is well-conducted, it is the fastest way to find treatments that work in people and ways to improve health. A clinical trial is one of the final stages of a long research process. It determines whether new experimental treatments or new ways of using known therapies are safe and effective, but it must meet ethical and scientific quality requirements. This paper reviews the principal phases of clinical trials and discusses the basic requirements needed to ensure a well-conducted experimental study. PMID- 16720364 TI - Profiling of hepatocellular proteins by 1D PAGE-MALDI/MS/MS in a rat heat stress model. AB - Heat induced complications cause an increase in a large number of proteins which play a role in diverse pathways during heat shock. A detailed characterization of these proteins is essential for understanding the molecular mechanisms involved in heat stroke. In this report, the proteins present in rat liver were compared at 37 degrees C (control) and at core temperature (Tc) 42 degrees C (heat stress) by 1D PAGE and MALDI/MS/MS. Among proteins identified in the sample after heat stress are dimethyglycine dehydrogenase, transketolase, carboxylic ester hydrolase, pyruvate kinase, L-type pyruvate kinase, arginosuccinate synthetase; fumarylacetoacetate hydrolase and peptidylpropyl isomerase A. These findings show that analysis of large scale proteins by MALDI/MS/MS provides a better understanding of the molecular mechanisms associated with heat shock. The resolution of proteins examined by 1D-PAGE was less than that obtained with 2D PAGE. More specifically, 2D-PAGE allows better identification of low molecular weight proteins that can not be resolved by 1D-PAGE. PMID- 16720365 TI - Negative regulation of T-cell receptor activation by the cAMP-PKA-Csk signalling pathway in T-cell lipid rafts. AB - Spatial organization of signal proteins in specialized cholesterol and glycosphingolipid-enriched microdomains (lipid rafts) provide specificity in lymphocyte signalling. Src kinases associate with lipid rafts on the basis of their dual acylation in the N-terminus and initiate T cell signalling. The immunomodulatory signal enzyme protein kinase A (PKA) is a serine/threonine kinase that controls a number of processes important for immune activation by phosphorylation of substrates that alters protein-protein interactions or changes the enzymatic activity of target proteins in T cells. PKA substrates involved in immune activation include transcription factors, members of the MAP kinase pathway, phospholipases and the Src kinase Csk. The PKA type I isoenzyme localizes to lipid rafts during T cell activation and modulates directly the proximal events that take place after engagement of the T cell receptor. The most proximal and major target for PKA phosphorylation is the C-terminal Src kinase Csk which initiates a negative signal pathway that fine-tunes the T cell activation process. PMID- 16720366 TI - Assessment of an anti-HIV-1 combination gene therapy strategy using the antisense RNA and multimeric hammerhead ribozymes. AB - A combination gene therapy strategy using an ASPsi-gag antisense RNA (targeted against the packaging signal and the gag-coding region) and a multimeric hammerhead ribozyme Rz1-9 (targeted against nine sites within the env-coding region) or Rz1-14 (targeted against 14 sites within the 5' leader and the pro-, pol-, vif- and env-coding regions) was assessed for inhibiting HIV-1 replication. A murine stem cell virus (MSCV)-based MGIN vector was used to express Rz1-9, Rz1 14, ASPsi-gag, Rz1-9ASPsi-gag, or Rz1-14ASPsi-gag RNA in a CD4+ T lymphoid cell line. Stable transductants were shown to express similar levels of interfering RNA. HIV-1 replication was inhibited in cells expressing Rz1-9 and Rz1-14. Little inhibition of HIV-1 replication was observed in cells expressing ASPsi-gag RNA. Thus, the multimeric hammerhead ribozymes inhibit HIV-1 replication better than the antisense RNA. Inhibition of HIV-1 replication in cells expressing Rz1-9ASPsi gag or Rz1-14ASPsi-gag RNA was worse than that obtained with the multimeric ribozymes alone. This result suggests that co-expression of antisense RNA decreases the anti-HIV potential of ribozymes. The multimeric ribozymes and the antisense RNA were designed to target different sites within the HIV-1 RNA. They are not expected to interact with each other. Neither are they expected to compete with each other for binding to the HIV-1 RNA. Instead, the antisense RNA binding to its (1553 nt-long) target site may have resulted in a decreased ribozyme turn over. Furthermore, since the antisense RNA/HIV-1 RNA hybrids are degraded by the cells, the co-expressed antisense RNA may have led to ribozyme degradation. PMID- 16720367 TI - Matrix metalloproteinases and their inhibitors in bone: an overview of regulation and functions. AB - Matrix metalloproteinases (MMPs), a family of Zn2+-dependent endopeptidases, mediate different physiological processes by digesting components of the extracellular matrix. Nevertheless, overexpression of MMPs is implicated in the pathogenesis of various diseases. Different MMPs and tissue inhibitors of MMPs (TIMPs) are expressed in bone cells, and their biosynthesis is regulated by local and systemic hormones and factors. The levels of enzymatically active MMPs in bone are further controlled by secretion, activation of proenzymes, inhibition by TIMPs, cellular uptake and degradation. Members of the cysteine and serine family of proteinases also coordinate some of the biological activities of MMPs in bone. The functions of MMPs and TIMPs in bone include regulation of processes, such as degradation of collagen and other components of the bone matrix, migration and survival of bone cells, endochondral bone formation and bone resorption. Abnormal expression of MMPs may lead to pathological conditions affecting bone and cartilage. Various pharmacological agents can inhibit MMPs, and some of these inhibitors may be potential therapeutic agents for certain bone diseases. This review briefly describes the regulation and functions of different MMPs and TIMPs in bone, and provides an insight into the role of MMPs in bone development, remodeling and pathology. PMID- 16720368 TI - Antisense delivery and protein knockdown within the intact central nervous system. AB - The ability to down regulate the expression of a specific protein within the intact central nervous system (CNS) is highly desirable from both a research and therapeutic perspective. Antisense has the potential to do this. However, problems of invasive antisense delivery methods and short half life of remain problematic. We overcome this by using Pluronic gel to provide a sustained delivery antisense oligodeoxynucleotides (ODN's) to the intact central nervous system and achieving rapid penetration throughout the spinal cord in 2-3 hours and significant knockdown of our target protein connexin 43 (Cx43) in 4-8 hours (recovering at 48-72 hours). Interestingly CY3-siRNA probes could not be detected penetrating the intact CNS and no knockdown the Cx43 was found. This approach with conventional ODNs could provide a faster and cheaper alternative to knockout mice in the investigation of the functions of specific proteins within the CNS and may also have therapeutic implications for drug discovery and development. PMID- 16720369 TI - A novel electrochemical biosensor for the detection of uric acid and adenine. AB - A novel electrochemical biosensor for the detection of uric acid and adenine was prepared based on a gel containing multi-walled carbon nanotubes and room temperature ionic liquid of 1-octyl-3-methylimidazolium hexafluorophosphate. The electrochemistry of uric acid and adenine was studied in this gel modified electrode. There was a significant two-way electrocatalytic activity upon both oxidation and reduction of uric acid. Similar to a bare glassy carbon electrode, uric acid undergoes a 2e,2H+ oxidation in phosphate buffer in the modified electrode. A diimine, the oxidation product of uric acid, was found to be an unstable intermediate, which was converted by a follow-up hydration reaction to an imine alcohol, with the reaction rate constant of 8.5 +/- 0.3 M(-1).s(-1) according to Nicholson's theory. Under optimum conditions, linear calibration graphs were obtained over the concentration range of 1.0 x 10(-7) M approximately 1.0 x 10(-5) M (uric acid) and 1.0 x 10(-5) M approximately 6.0 x 10(-4) M (adenine). Based on the signal-to-noise ratio of 3, the detection limits of the current technique was found to be as low as 9.0 x 10(-8) M (uric acid) and 2.0 x 10(-6) M (adenine), respectively. This novel biosensor was successfully applied for the assay of uric acid in human urine. Because of its good stability and long term durability, such a gel modified electrode can provide a simple and easy approach for sensitive detection of uric acid and adenine. PMID- 16720370 TI - Spiroplasmas: evolutionary relationships and biodiversity. AB - Spiroplasmas are wall-less descendants of Gram-positive bacteria that maintain some of the smallest genomes known for self-replicating organisms. These helical, motile prokaryotes exploit numerous habitats, but are most often found in association with insects. Co-evolution with their insect hosts may account for the highly speciose nature of the genus Spiroplasma, with many spiroplasmas existing in obligate insect/plant transmission cycles. In addition to insect and plant hosts, spiroplasmas are found in association with ticks and crustaceans. Although most spiroplasma associations appear to be commensal, some cases of pathogenicity or mutualism have been described. Most notably, spiroplasmas have been identified as the causative agents of agricultural and aquacultural diseases and the sex ratio disorder in insects. Some spiroplasmas exhibit strict host and/or geographical ranges, but others are relative generalists. Species of the genus Spiroplasma have been traditionally classified into 34 groups based on cross-reactivity of surface antigens. Three of the serogroups contain closely related strain complexes that are further divided into subgroups. Phylogenetic reconstructions based on 16S rDNA sequence strongly support the closely related serogroups. To date, less than 40 Spiroplasma species have been fully characterized and given binomial names. Complete characterization of a new species involves numerous phenotypic and genotypic tests as outlined in the minimal standards document; this document is currently under revision to include phylogenetic data and a reevaluated set of required phenotypic and genotypic tests. The area of spiroplasma research is poised for major advances with new criteria for naming species in preparation, a dramatic increase in available molecular characters, the promise of full genome sequences, and advances in genetic tools for manipulation of these organisms. PMID- 16720371 TI - Recent advances in melanoma research. AB - At the end of the last decade, sporadic melanomas were still considered a genetic black box. Fortunately, in the last few years the box has been opened bringing to light melanoma-relevant oncogenes, aberrant signal transduction pathways, critical alterations in the melanoma cell cycle that go beyond p16INK4a, and melanoma- microenvironment interactions that are essential for tumor progression. This review will discuss some of the latest findings in melanoma research including the critical role of the MAPK pathway in the genesis of melanoma and senescence of nevi, the paradoxical tumor suppressor and oncogenic activities of the transcription factor MITF, and the unexpected oncogenic activities of the low molecular weight forms of cyclin E. PMID- 16720372 TI - Connexin 43 gene expression in mice with cardiopulmonary developmental defects. AB - Gap junctions are vital for cellular integrity, including homeostasis, morphogenesis, differentiation and growth in normal development of organs such as heart. Connexin 43 (Cx43) is a major gap junction protein. Our cDNA microarray analysis of normal and nitrofen-exposed neonatal mice with hypoplastic lungs, associated congenital diaphragmatic hernia (CDH) and heart developmental defects showed up-regulation of Cx43. Our objective was to establish if cardiopulmonary defects in nitrofen-exposed mice may be linked to altered expression of the Cx43 gene. We addressed our objective by performing northern blot analysis, real-time RT-PCR, immunoblotting and immunohistochemistry by localizing Cx43 in hearts and lungs of normal and nitrofen-exposed mice at different gestational stages. The data confirmed up-regulation of Cx43 expression in both hearts and lungs of CDH neonate mice and in lungs at other developmental stages except the pseudoglandular stage. However, Cx43 protein levels were either the same or less in hearts and lungs of nitrofen-exposed mice than in normal tissues except in pseudoglandular lungs. Different expressions of mRNA and protein suggest possible post-transcriptional or translational defects in Cx43. We observed dysmorphic hearts with exaggerated interventricular grooves and deep notches at the apex of the hearts in nitrofen-exposed fetal/neonatal mice; narrowed pulmonary out-flow and various degrees of craniofacial defects in 15-20% of the affected mice. Our data suggest a possible involvement of Cx43 in craniofacial, heart and lung defects in nitrofen-exposed mice. Such cardiopulmonary defects are also observed in human newborns with CDH. Thus, the murine data may help elucidate the pathways of cardiopulmonary defects in the human newborn condition. PMID- 16720373 TI - Transglutaminase 2 in inflammation. AB - Many reports have shown that the expression of transglutaminase 2 (TG 2) is increased in inflammatory diseases. Although during the last several decades multiple physiological roles for TG 2 have been demonstrated in various cell types, its role in the inflammatory process is not yet clear. TG 2 is a crosslinking enzyme that is widely used in many biological systems for tissue stabilization purposes and immediate defense against injury or infection. Aberrant activation of TG 2 activity in tissues contributes to a variety of diseases including neurodegenerative diseases, autoimmune diseases, and cancers. In most cases, TG 2 appears to form an inappropriate protein aggregate that may be cytotoxic enough to trigger inflammation and/or apoptosis. In some cases, such as celiac disease and rheumatoid arthritis, TG 2 is also associated with the pathogenic progression, as well as in the generation of autoantibodies. Recently, we discovered that increased TG 2 activity triggers NF-kappaB activation without I-kappaBalpha kinase signaling. TG 2 induces the polymerization of I-kappaBalpha rather than stimulating I-kappaBalpha kinase. This polymerization of I-kappaB results in the direct activation of NF-kappaB in various cell lines. We also found that TG inhibition reverses NF-kappaB activation. Interestingly, this coincides with the reversal of inflammation in conjunctivitis models by treatment with TG 2 inhibitors. Here, I introduce a new role for TG 2 as a signal modulator, which may suggest a new paradigm for the inflammatory process. PMID- 16720374 TI - Mechanisms of doxycycline-induced cytotoxicity on human bronchial epithelial cells. AB - Doxycycline (DOX), a synthetic tetracycline, may have potential utility in the management of cancers and in the treatment of chronic inflammatory diseases due to its role in growth, invasion and metastasis of many tumors, on cell proliferation and as inducer of apoptosis. Some studies established its role in the treatment of lesions induced by mustards, warfare agents causing severe damage with blistering and tissue detachment in exposed areas of the body. In the present study, the effect of Dox was investigated in a human bronchial epithelial cell line. Dox induced a time- and concentration-dependent cell proliferation inhibition, associated with a cell cycle arrest in S phase, a decrease in viability due to apoptosis and necrosis, and cell detachment. This latter was partly correlated with early activation of caspase-3 before detachment, and with mitochondrial alteration. Cell transfection with a Bcl-2 encoding vector showed a decrease both in mitochondrial depolarization and cell detachment. Dox-induced apoptosis included decrease in Bcl-2 expression, increase in Bak expression and caspase-3 and -9 activation but appeared to be p53- and Bax-independent. A better comprehension of the Dox-induced apoptotic pathway could allow to abolish its toxic effects, improving the therapeutic efficiency of Dox. PMID- 16720375 TI - Y chromosome and male infertility: update, 2006. AB - Male factor infertility accounts for about half the cases of couple infertility and in around 50% of cases its etiology remains unknown. Molecular genetic techniques have unveiled a number of etiopathogenetic factors, including microdeletions of the Yq. Y chromosome microdeletions removing the AZoospermia Factor (AZF) regions are the most frequent molecular genetic causes of oligo/azoospermia. The intense effort of many laboratories contributed to a better understanding of the clinical significance of this genetic anomaly and to the identification of fertility candidate genes in the AZF regions. Important progress has been made on the structure of the Y chromosome and the mechanism of deletion. Studies aimed to define a predisposing genetic background for Yq deletions were not successful, perhaps due to the low number of patients analyzed so far. The screening for Yq deletions became a routine diagnostic test that provides an etiology for spermatogenic disturbances, and assess in the prognosis for testicular sperm retrieval according to the type of deletion. Assisted reproductive techniques represent an efficient symptomatic therapy for men bearing Y microdeletions, however, this genetic defect is transmitted to the male offsprings, affecting their fertility. Future studies should focus on understanding the biological function of AZF genes which is an essential step for the development of more appropriate and knowledge-based therapies. PMID- 16720376 TI - MutY and MutY homologs (MYH) in genome maintenance. AB - The base excision repair carried out by bacterial MutY DNA glycosylase and eukaryotic MutY homolog (MYH) is responsible for removing adenines misincorporated into DNA opposite G and 7,8-dihydro-8-oxo-guanines (8-oxoG); thereby preventing G:C to T:A mutations. Escherichia coli MutY (EcMutY) can also remove adenines from A/C and A/5-hydroxyuracil and can remove guanines from G/8 oxoG mismatches at reduced rates. Thus, MutY has a minor role in reducing the mutagenic effects on G:C to A:T transitions and G:C to C:G transversions. The eukaryotic MYH can excise adenines misincorporated opposite GO, G, or C; remove 2 hydroxyadenines mispaired with A,G, and GO; excise G from G/GO mismatch weakly, thereby preventing G:C to T:A transversions. The in vitro and in vivo activities of MYH can be modulated by several proteins including apurinic/apyrimidinic endonuclease (APE1), proliferating cell nuclear antigen (PCNA), and mismatch recognition enzymes MSH2/MSH6. Recently, MYH has been shown to associate with the checkpoint proteins, Rad9, Rad1, and Hus1 (referred as the 9-1-1 complex). Thus, MYH-mediated base excision repair is coordinated with mismatch repair, DNA replication, cell-cycle progression, and DNA-damage checkpoints. Biallelic germ line mutations in the human MYH gene are associated with recessive inheritance of multiple colorectal adenomas and carcinoma. MYH mutations can cause G:C to T:A mutations of the adenomatous polyposis coli (APC), K-ras, and other genes that control cellular proliferation in the colon. PMID- 16720377 TI - The role of collagenolytic matrix metalloproteinases in the loss of articular cartilage in osteoarthritis. AB - Digestion of cartilage collagen is a critical step in the loss of articular cartilage in osteoarthritis (OA). The hypothesis that matrix metalloproteinases (MMPs) are the primary enzymes involved in cartilage collagen digestion is supported by evidence that indicates that: 1) one or more MMP can digest the extracellular matrix components in vitro, 2) the enzymes are expressed in OA cartilage at the place and time of cartilage destruction, 3) specific digestion products of MMP are present in the OA cartilage, and 4) experimental strategies that alter the expression and/or the activity of MMP alter the progression of cartilage destruction in OA. These observations suggest that MMPs in general and collagenolytic MMPs in particular are promising targets for treatment of OA. PMID- 16720378 TI - Detection of predefecatory rectosigmoid wave activity for prevention of fecal soiling in infants. AB - Identification of an electrophysiologic sign before defecation can prevent fecal soiling in infants. To identify such a sign, the contractile activity of sigmoid colon was recorded percutaneously in 48 healthy infants. The recorder was equipped with a digital clock synchronized to the recorder so as to set off an alarm upon significantly increased electromyographic activity of sigmoid colon. Examination of the recordings at high speed revealed three types of basal, signaling and predefecatory waves of activities. The 'basal' component was comprised of as negatively deflected slow waves. The signaling waves exhibited an increase in amplitudes, cycle rate and conduction velocity, were repeated 8.2+/ 1.2 times and lasted for 14.6+/-2.1 minutes prior to defecation, The 'predefecatory' waves preceded defecation by 40.3+/-7.3 seconds, showed a significant increase in wave parameters and sounded the alarm. The findings show a method for early detection of defecation that can be used clinically to prevent fecal soiling in infants. PMID- 16720379 TI - Matrix metalloproteinase dependent and independent collagen degradation. AB - Cleavage of the fibrillar collagens occurs during physiological conditions, as well as pathological conditions. The resistance of the fibrillar collagens to degradation is due to their rigid and compact structures. There are only a limited number of proteinases that have the capability to initiate the cleavage of the fibrillar collagens. These include some of the matrix metalloproteinases (MMPs) and cathepsins, as well as a few serine proteinases. The MMPs have long been implicated in the collagen degradation and remodeling that occurs at physiological pHs. The cathepsins, on the other hand, have been implicated in the collagen cleavage that occurs at acidic pHs, particularly the collagen degradation that is mediated by osteoclasts. In addition to the MMPs, a few serine proteinases have been implicated in the collagen degradation that occurs at neutral pH. The characteristics that contribute to the resistance of the fibrillar collagens to cleavage are discussed along with the MMPs, cathepsins, and serine proteinases that can cleave these collagens. PMID- 16720381 TI - NAD+ and NADH in cellular functions and cell death. AB - Increasing evidence has indicated that NAD+ and NADH play critical roles not only in energy metabolism, but also in cell death and various cellular functions including regulation of calcium homeostasis and gene expression. It has also been indicated that NAD+ and NADH are mediators of multiple major biological processes including aging. NAD+ and NADH produce the biological effects by regulating numerous NAD+/NADH-dependent enzymes, including dehydrogenases, poly(ADP-ribose) polymerases, Sir2 family proteins (sirtuins), mono(ADP-ribosyl)transferases, and ADP-ribosyl cyclases. Of particular interest, NAD+-dependent generation of ADP ribose, cyclic ADP-ribose and O-acetyl-ADP-ribose can mediate calcium homeostasis by affecting TRPM2 receptors and ryanodine receptors; and sirtuins and PARPs appear to play key roles in aging, cell death and a variety of cellular functions. It has also been indicated that NADH and NAD+ can be transported across plasma membranes of cells, and that extracellular NAD+ may be a new signaling molecule. Our latest studies have shown that intranasal NAD+ administration can profoundly decrease ischemic brain damage. These new pieces of information have fundamentally changed our understanding about NAD+ and NADH, suggesting novel paradigms about the metabolism and biological activities of NAD+ and NADH. Based on this information, it is tempted to hypothesize that NAD+ and NADH, together with ATP and Ca2+, may be four most fundamental components in life, which can significantly affect nearly all major biological processes. Future studies on NAD+ and NADH may not only elucidate some fundamental mysteries in biology, but also provide novel insights for interfering aging and many disease processes. PMID- 16720380 TI - MMP-9 expression is associated with leukocytic but not endothelial markers in brain arteriovenous malformations. AB - Brain arteriovenous malformations (BAVM) have high matrix metalloproteinase-9 (MMP-9) expression, the source of which is unclear. We hypothesized MMP-9 production might be due to inflammation in BAVM. Compared to control brain tissues (n = 5), BAVM tissue (n = 139) had a higher expression (by ELISA) of myeloperoxidase (MPO) (193 +/- 189 vs. 6 +/- 3, ng/mg, P < .001), MMP-9 (28 +/- 32 vs. 0.7 +/- 0.6, ng/mg, P < .001), and IL-6 (102 +/- 218 vs. 0.1 +/- 0.1, pg/mg, P < .001), but not eNOS (114 +/- 87 vs. 65 +/- 9, pg/mg, P = .09). MMP-9 expression in BAVM highly correlated with myeloperoxidase (R2 = .76, P < .001), as well as with IL-6 (R2 = .32, P < .001). In contrast, MMP-9 in BAVM poorly correlated with the endothelial marker, eNOS (R2 = .03, P = .05), and CD31 (R2 = .004, P = .57). Compared to non-embolized patients (n = 46), patients with pre operative embolization (n = 93) had higher levels of myeloperoxidase (236 +/- 205 vs. 106 +/- 108, ng/mg, P < .001) and MMP-9 (33 +/- 35 vs. 16 +/- 20, ng/mg, P < .001), however the correlation between MMP-9 and myeloperoxidase was equally strong for both groups (R2 = .69, n = 93, P < .001, for both). MMP-9 expression correlated with the lipocalin-MMP-9 complex, suggesting neutrophils as the MMP-9 source. MPO co-localized with majority of MMP-9 signal by immunohistochemistry. Our data suggest that inflammation is a prominent feature of BAVM lesional phenotype, and neutrophils appear to be a major source of MMP-9 in these lesions. PMID- 16720382 TI - Magnesium transporters: properties, regulation and structure. AB - The chemistry of Mg2+ is unique amongst biological cations, and the properties of Mg2+ transport systems reflect this chemistry. Prokaryotes carry three classes of Mg2+ transport systems: CorA, MgtA/B and MgtE. CorA and MgtE are widely distributed in both Eubacteria and Archaea, while the MgtA/B class is found primarily in the Eubacteria. Eukaryotic homologs of CorA, although clearly functional as Mg2+ transporters, have minimal sequence homology and include the Mrs2p mitochondrial Mg2+ channel and the ALR proteins of fungi. MgtE homologs are more recognizable in eukaryotes as the SLC41 class of transporters. The MgtA/B Mg2+ transporters belong to the P-type ATPase superfamily, but mediate Mg2+ influx down its electrochemical gradient rather than against the gradient as with other P-type ATPases. Their physiological role is not clear. CorA is the only Mg2+ transporter whose structure has been solved. It is a homopentamer with two transmembrane domains per monomer, the first of which forms the ion conduction pathway. Mg2+ transport involves first the binding of the fully hydrated cation to an extracellular binding loop connecting the transmembrane domains. Passage through the membrane involves no electrostatic interactions, but two cytosolic domains, one carrying extremely high concentrations of positive charge and the other negative charge appear to help control Mg2+ flux, in concert with an intracellular Mg2+ bound between domains of each monomer. Neither CorA nor MgtE appear to be transcriptionally regulated, implying they are primarily "housekeeping" genes. Nonetheless, mutation of the corA gene in Salmonella enterica serovar Typhimurium leads to attenuation of virulence and other defects, even though the strain carries two additional Mg2+ transporters and the mutant exhibits no Mg2+-dependent growth deficit. PMID- 16720383 TI - Pathways from glutamine to apoptosis. AB - It is fascinating that a relatively simple amino acid like glutamine is involved in such large variety of cellular reactions. Glutamine is required for nitrogen stimulated proliferation in many cells, but glutamine stimulates not only the growth of cells but also the expression of surface antigens, the formation of cytokines, the synthesis of heat shock proteins and many more vital events. Among all of them, apoptosis is a recent but outstanding incorporation to the whole of phenomena regulated by this peculiar amino acid. Apoptosis is an important process in a wide number of biologic systems. Apoptotic signalling mechanisms implicated in response to glutamine deprivation are cell type-specific. In any case, new findings indicate that glutamine availability is strongly related to the induction of apoptosis, working both as a nutrient and as a signalling molecule, acting directly or indirectly on the pathways leading to programmed cell death. Following, we will describe as glutamine and the related species glutamate, glutathione and glucosamine can play important roles in the pathways leading to apoptosis. PMID- 16720384 TI - The interaction of G-CSF with its receptor. AB - The function of the G-CSF receptor has been of considerable interest, particularly because of the clinical usefulness of G-CSF. The first step in receptor activation, which is the interaction of G-CSF with its receptor, has been studied by mapping the binding sites of neutralizing antibodies, by studying the complexes formed between G-CSF and various receptor fragments in solution and by mutagenesis of the receptor and ligand. In addition, the structure of G-CSF has been determined. Part of the ligand-binding domain of the receptor in complex with G-CSF has been crystallized and its structure described. Consideration of all these studies has allowed us to make a model of the complete ligand-binding domain in complex with G-CSF that accounts for the published data. The complex has a 2:2 stoichiometry, with two binding sites on both the ligand and receptor that are equivalent to site II and site III of the IL-6 receptor complex. This model was based on the published structure of gp130 in complex with viral IL-6, which we believe to be very similar. PMID- 16720385 TI - Recombinant adeno-associated viral vector encoding human VEGF165 induces neomicrovessel formation in the adult mouse brain. AB - Delivery of therapeutic genes represents a fascinating possibility to accelerate injury-repairing process in tissues that are otherwise difficult to treat, such as cerebral ischemia. Current studies indicate that gene transfer-induced focal angiogenesis in the brain may provide an important therapeutic strategy. In the present study, we reported the efficacy of induction of angiogenesis with an adeno-associated virus (AAV) vector expressing the 165 amino acid isoform of vascular endothelial growth factor (VEGF165). We found AAV serotype 1 has more efficiency in transduction of the brain tissue than AAV serotype 2. Quantitative vessel counting showed that microvessels in AAV-VEGF transduced mice significantly increased from 1 week up to 12 weeks compared to the control groups (AAV-VEGF: 316+/-58 vs. AAV-lacZ: 180+/-34 and saline: 152+/-35 vessels/mm2, at 6 weeks, p<0.05). Proliferating cell nuclear antigen (PCNA) staining confirmed these microvessels were actively proliferating. Double-labeled fluorescence staining demonstrated that neurons, astrocytes, and endothelial cells could express VEGF following AAV-VEGF gene transfer. AAV vectors did not elicit a detectable inflammatory response, cell loss or neuronal damage. Our data underline the importance of angiogenesis in the brain tissue and indicate that VEGF gene transfer might present a valuable approach to treat brain ischemic disorders. PMID- 16720386 TI - Adaptive cellular mechanisms in response to glutamine-starvation. AB - Glutamine (Gln) utilising cells suffer from Gln-starvation during critical illness when plasma Gln levels are decreased. This study investigates whether such cells activate adaptive mechanisms. Monocytic U937 cells were cultured at 0.6 and 0.2 mM Gln for up to four days. Within the first day a decrease of ATP (78% of control), intracellular free Gln (13%), Hsp70 (74%) and proliferation rate (79%) was observed. A prolonged culture at 0.6 mM Gln for additional three days led to a recovery of ATP (97%), Hsp70 (91%) and proliferation (92%). The intracellular free Gln increased only to 41%. At 0.2 mM Gln, however, all levels remained decreased. The activation of the metabolic sensor AMP activated kinase (AMPK) increased immediately in Gln-starving cells but regained normal values only in cells cultured at 0.6 mM. A proteomic analysis identified 23 proteins, which were affected by Gln starvation including metabolic enzymes, proteins involved in synthesis and degradation of RNA and proteins, and stress proteins. These data show that Gln-utilising cells activate adaptive mechanisms in response to Gln-starvation, which enable them to overcome a Gln shortage. At very low Gln concentrations, these adaptive mechanisms are not sufficient to countervail the lack of the amino acid. PMID- 16720387 TI - Identification of an octamer-binding site controlling the activity of the small breast epithelial mucin gene promoter. AB - e human small breast epithelial mucin (SBEM) gene has been identified as being preferentially expressed in mammary epithelial cells and over-expressed in breast tumors. In this report, we have characterized the promoter of SBEM gene in order to identify sequences responsible for this strong mammary expression. A series of SBEM promoter/luciferase constructs were transiently transfected into both breast (MCF-7, BT-20) and non-breast (HeLa and HepG2) cell lines. In addition to the minimal promoter and to a repressor region, we have identified an 87-bp sequence (-357/-270) driving a strong breast-specific expression. Site-directed mutagenesis of a putative octamer-binding transcription factor binding site located within this latter region led to a strong decrease of the transcriptional activity of the SBEM promoter. Furthermore, transient over-expression of Oct1 and Oct2 not only increased SBEM promoter reporter activity, but also enhanced endogenous SBEM mRNA level. Overall, the data suggest that octamer-binding transcription factors participate in the strong expression of SBEM gene in breast tissues. Clarifying the SBEM gene regulation will help to dissect mechanisms underlying transcription of normal breast and breast cancer-associated genes. PMID- 16720388 TI - Complex population dynamics and complex causation: devils, details and demography. AB - Population dynamics result from the interplay of density-independent and density dependent processes. Understanding this interplay is important, especially for being able to predict near-term population trajectories for management. In recent years, the study of model systems-experimental, observational and theoretical-has shed considerable light on the way that the both density-dependent and independent aspects of the environment affect population dynamics via impacting on the organism's life history and therefore demography. These model-based approaches suggest that (i) individuals in different states differ in their demographic performance, (ii) these differences generate structure that can fluctuate independently of current total population size and so can influence the dynamics in important ways, (iii) individuals are strongly affected by both current and past environments, even when the past environments may be in previous generations and (iv) dynamics are typically complex and transient due to environmental noise perturbing complex population structures. For understanding population dynamics of any given system, we suggest that 'the devil is in the detail'. Experimental dissection of empirical systems is providing important insights into the details of the drivers of demographic responses and therefore dynamics and should also stimulate theory that incorporates relevant biological mechanism. PMID- 16720389 TI - Critique of directionality theory. PMID- 16720390 TI - Alteration of the chemical environment disrupts communication in a freshwater fish. AB - It is well established that changes to the chemical environment can impair development, physiology and reproductive biology; by contrast, impacts on communication have not been widely reported. This is surprising given that chemical communication is the most widely used sensory modality in nature, and that variation in the chemical composition of the environment is the rule, not the exception. Here, we show that chemically mediated species recognition in a swordtail fish, Xiphophorus birchmanni, can be hindered by anthropogenic disturbance to the signalling environment. Females have a strong preference for conspecific male chemical cues, yet they hybridize in nature with the congener X. malinche. Wild-caught females showed a strong preference for conspecifics when tested in clean water, but failed to show a preference when tested in stream water subject to sewage effluent and agricultural runoff. We hypothesized that this was due to the interaction between chemical communication systems and humic acid (HA), a ubiquitous, natural product elevated to high levels by anthropogenic processes. When exposed to elevated concentrations of HA, female X. birchmanni again lost their preference for conspecific male chemical cues, while visual mating preferences and motivation to mate were retained. Sub-lethal concentrations of seemingly benign substances can thus have a drastic effect on natural populations through their specific impact on communication systems. PMID- 16720391 TI - Encounter success of free-ranging marine predator movements across a dynamic prey landscape. AB - Movements of wide-ranging top predators can now be studied effectively using satellite and archival telemetry. However, the motivations underlying movements remain difficult to determine because trajectories are seldom related to key biological gradients, such as changing prey distributions. Here, we use a dynamic prey landscape of zooplankton biomass in the north-east Atlantic Ocean to examine active habitat selection in the plankton-feeding basking shark Cetorhinus maximus. The relative success of shark searches across this landscape was examined by comparing prey biomass encountered by sharks with encounters by random-walk simulations of 'model' sharks. Movements of transmitter-tagged sharks monitored for 964 days (16754 km estimated minimum distance) were concentrated on the European continental shelf in areas characterized by high seasonal productivity and complex prey distributions. We show movements by adult and sub adult sharks yielded consistently higher prey encounter rates than 90% of random walk simulations. Behavioural patterns were consistent with basking sharks using search tactics structured across multiple scales to exploit the richest prey areas available in preferred habitats. Simple behavioural rules based on learned responses to previously encountered prey distributions may explain the high performances. This study highlights how dynamic prey landscapes enable active habitat selection in large predators to be investigated from a trophic perspective, an approach that may inform conservation by identifying critical habitat of vulnerable species. PMID- 16720392 TI - Termite-egg mimicry by a sclerotium-forming fungus. AB - Mimicry has evolved in a wide range of organisms and encompasses diverse tactics for defence, foraging, pollination and social parasitism. Here, I report an extraordinary case of egg mimicry by a fungus, whereby the fungus gains competitor-free habitat in termite nests. Brown fungal balls, called 'termite balls', are frequently found in egg piles of Reticulitermes termites. Phylogenetic analysis illustrated that termite-ball fungi isolated from different hosts (Reticulitermes speratus, Reticulitermes flavipes and Reticulitermes virginicus) were all very similar, with no significant molecular differences among host species or geographical locations. I found no significant effect of termite balls on egg survivorship. The termite-ball fungus rarely kills termite eggs in natural colonies. Even a termite species (Reticulitermes okinawanus) with no natural association with the fungus tended termite balls along with its eggs when it was experimentally provided with termite balls. Dummy-egg bioassays using glass beads showed that both morphological and chemical camouflage were necessary to induce tending by termites. Termites almost exclusively tended termite balls with diameters that exactly matched their egg size. Moreover, scanning electron microscopic observations revealed sophisticated mimicry of the smooth surface texture of eggs. These results provide clear evidence that this interaction is beneficial only for the fungus, i.e. termite balls parasitically mimic termite eggs. PMID- 16720393 TI - Small object detection neurons in female hoverflies. AB - While predators such as dragonflies are dependent on visual detection of moving prey, social interactions make conspecific detection equally important for many non-predatory insects. Specialized 'acute zones' associated with target detection have evolved in several insect groups and are a prominent male-specific feature in many dipteran flies. The physiology of target selective neurons associated with these specialized eye regions has previously been described only from male flies. We show here that female hoverflies (Eristalis tenax) have several classes of neurons within the third optic ganglion (lobula) capable of detecting moving objects smaller than 1 degrees . These neurons have frontal receptive fields covering a large part of the ipsilateral world and are tuned to a broad range of target speeds and sizes. This could make them suitable for detecting targets under a range of natural conditions such as required during predator avoidance or conspecific interactions. PMID- 16720394 TI - Attentional modulation of perceptual stabilization. AB - Perceptual priming is generally regarded as a passive and automatic process, as it is obtained even without awareness of the prime. Recent studies have introduced a more active form of perceptual priming in which priming for a subsequent ambiguous stimulus is triggered by the subjective percept, that is, interpretation of a previous ambiguous stimulus. This phenomenon known as stabilization does not require a conscious effort to actively maintain one perceptual interpretation. In this study, we show that distraction of attention, during and even after the prime presentation, interferes with the build-up of perceptual memory for stabilization. This implies that despite the apparent automaticity, stabilization involves an active attentional process for encoding and retention. The disruption during the encoding can be attributed to the reduction in sensory signals for the prime. However, the disruption during the retention suggests that the implicit memory trace of the prime necessitates the attentional resource to fully develop. The active nature of the build-up of perceptual memory for stabilization is consistent with the idea that perceptual memory increases its strength gradually over a few seconds. These findings suggest that seemingly automatic and effortless cognitive processes can compete with online perceptual processing for common attentional resources. PMID- 16720395 TI - Are capuchin monkeys (Cebus apella) inequity averse? AB - It has been reported that capuchin monkeys reject a less preferred food (LPF) when they see a partner capuchin receive a more preferred food (PF) for performing the same task. This behaviour was taken as evidence of 'inequity aversion', but an alternative hypothesis is that capuchins reject the LPF because of the mere presence of the PF. We tested this hypothesis in a paradigm, which consisted of presenting two different foods (one PF and one LPF) on a tray and allowing the capuchin to take only the LPF. Refusals to initiate the trial and refusals to take and eat the LPF were higher when the PF was hidden (hiding condition) and when the PF was accumulated in sight but out of reach of the subject (accumulation condition) compared to when two pieces of LPF were placed on the tray (control condition). Interestingly, the subject behaved as in the control condition when its partner was given and ate the PF (partner condition). We argue that capuchins' refusals were due to the frustration of seeing and not obtaining the PF, and that seeing the partner eating increases the LPF acceptance. PMID- 16720396 TI - Age-structured effects and disease interference in childhood infections. AB - Recent studies have demonstrated that ecological interference among some childhood diseases may have important dynamic consequences. An interesting question is, when would we expect the interference effect to be pronounced? To address the issue, here we develop a seasonally forced two-disease age-structured model, using empirically derived age-specific force of infection (ASFOI) for numerous infections of childhood. Our comparative numerical analysis shows that when the ASFOIs for the two diseases largely overlap, the dynamics predicted by the two-disease model are generally different from those predicted by the analogous single-disease model, suggesting strong fingerprints of disease interference. When the ASFOIs overlap less, on the other hand, both diseases behave as predicted by the single-disease model, suggesting weak interference. We conclude that age structure is an important factor that should be taken into account in order to explore the underlying mechanisms of disease interference. PMID- 16720397 TI - Good reindeer mothers live longer and become better in raising offspring. AB - Longevity is the main factor influencing individual fitness of long-lived, iteroparous species. Theories of life history evolution suggest this is because increased longevity allows individuals to (i) have more breeding attempts (time component), (ii) accumulate experience so as to become better able to rear offspring (experience component) or (iii) because individuals reaching old age have above-average quality (quality component). We assess empirically the relative influences of time, experience and quality on the relationship between longevity and individual fitness among female reindeer. Fitness increased with longevity due to all three processes. All females increased in success with age up to their penultimate year of life (experience component), the success of the terminal-breeding occasion was strongly dependent on longevity. Long-lived females had more successful breeding attempts during their life (time component), and had higher reproductive success at all ages, especially during the last year of life (individual quality component) than short-lived females. Our study reveals a more complex relationship between longevity and fitness in large mammals than the simple increase of the number of reproductive attempts when living longer. PMID- 16720398 TI - Barn swallows before barns: population histories and intercontinental colonization. AB - The barn swallow (Hirundo rustica) is one of most widely distributed swallows, owing in part to its recent switch from natural nest sites to human structures. We conducted phylogenetic analysis of mitochondrial (mt) and nuclear DNA to explore the recent evolutionary history of this species. Strongly supported mtDNA clades corresponded to Europe, Asia and North America plus the Baikal region of Asia. Analysis of sequence data from a sex-linked nuclear gene was unable to recover the phylogenetic splits in the mtDNA tree, confirming that the main clades evolved recently. The phylogenetic pattern suggests that the ancestral area of the barn swallow was the holarctic; most divergence events are consistent with vicariance. Most unexpectedly, analyses show that barn swallows from North America colonized the Baikal region in the recent past (one fixed substitution). This dispersal direction is opposite of that for most nearctic-palearctic taxon exchanges. Although this invasion was envisioned to coincide with the appearance of new types of human dwelling in the Baikal region, calibration of molecular divergence suggests an older dispersal event. A recent history of gene flow within the main palearctic clades is consistent with range and population expansion owing to new nesting opportunities provided by human settlements. Contrary to expectation, populations in North America appear historically larger and more stable than those in the palearctic. The Baikal population apparently has not increased greatly since colonization. PMID- 16720399 TI - Fitness landscapes support the dominance theory of post-zygotic isolation in the mussels Mytilus edulis and M. galloprovincialis. AB - We studied the genetic basis of post-zygotic isolation in the marine mussels Mytilus edulis and Mytilus galloprovincialis. Evidence was obtained for a high number of recessive Dobzhansky-Muller substitutions in the genome of these two mussel taxa. We analysed the segregation of unlinked diagnostic markers in the progeny of two backcrosses and an F2 cross, 36 h and 200 days after fertilization. Directional selection favouring M. galloprovincialis genotypes was observed in both kinds of cross. In the F2, epistatic interactions between each pair of chromosome fragments mapped by the markers were identified in addition. Our results imply that homozygous-homozygous interactions are required for breakdown of coadaptation, in accordance with the dominance theory of post zygotic isolation. Endogenous post-zygotic selection distributed over many loci throughout the genome provides the missing factor explaining the astonishing persistence and strength of barriers to neutral introgression in such a dispersive taxon as Mytilus. PMID- 16720400 TI - Alien mink predation induces prolonged declines in archipelago amphibians. AB - Amphibians are undergoing enigmatic global declines variously attributed to a complex web of anthropogenic forces. Alien predators pose a fundamental threat to biodiversity generally that is predicted to be most acute in island ecosystems. While amphibian eggs and tadpoles are vulnerable to aquatic predators, the effect of predators on adult, reproducing frogs, which most influence amphibian population processes, is unknown. Here, we report on the responses of amphibian populations in the outer Finnish Archipelago to a long-term, large-scale removal of American mink (Mustela vison Schreb.), an invasive predator linked to recent biodiversity loss across Europe. Removal increased both the densities and distribution of common frogs (Rana temporaria L.) but not those of common toads (Bufo bufo L.), which appear to escape mink predation because of their unpalatable skin. Importantly, the largest benefits of mink removal to frog recovery were slow to appear as frogs apparently have a delayed maturation in these harsh environments, which means we must be cautious about reliance upon short-term results. PMID- 16720401 TI - The predation costs of symmetrical cryptic coloration. AB - In psychological studies of visual perception, symmetry is accepted as a potent cue in visual search for cryptic objects, yet its importance for non-human animals has been assumed rather than tested. Furthermore, while the salience of bilateral symmetry has been established in laboratory-based search tasks using human subjects, its role in more natural settings, closer to those for which such perceptual mechanisms evolved, has not, to our knowledge, been investigated previously. That said, the salience of symmetry in visual search has a plausible adaptive rationale, because biologically important objects, such as prey, predators or conspecifics, usually have a plane of symmetry that is not present in their surroundings. We tested the conspicuousness to avian predators of cryptic artificial, moth-like targets, with or without bilateral symmetry in background-matching coloration, against oak trees in the field. In two independent experiments, symmetrical targets were predated at a higher rate than otherwise identical asymmetrical targets. There was a small, but significant, fitness cost to symmetry in camouflage patterns. Given that birds are the most commonly invoked predators shaping the evolution of defensive coloration in insects, this raises the question of why bilateral asymmetry is not more common in cryptic insects. PMID- 16720403 TI - The cost of catching up: increased winter mortality following structural growth compensation in the wild. AB - Although laboratory and observational studies suggest that many animals are capable of compensatory growth after periods of food shortage, few field experiments have demonstrated structural growth compensation in the wild. Here, we addressed the hypotheses that (i) food restriction can induce structural compensatory growth in free-living animals, (ii) that compensation is proportional to the level of body size retardation and (iii) that compensation induces mortality costs. To test these, wild brown trout (Salmo trutta) yearlings were brought to the lab, tagged individually, subjected to four levels of food deprivation (including a control), released back into the native stream and recaptured after one, five and ten months. Brown trout fully restored condition and partially restored mass within a month, whereas compensation in structure (i.e. body length) was not evident until after five months, supporting hypothesis 1. As the level of growth compensation was similar among the three deprived groups, hypothesis 2 was not supported. A final recapture after winter revealed delayed mortality, apparently induced by the compensatory response in the deprived groups, which is consistent with hypothesis 3. To our knowledge, this is the first field experiment demonstrating structural compensatory growth and associated costs in a wild animal population. PMID- 16720402 TI - Costs and benefits of a superinfection of facultative symbionts in aphids. AB - Symbiotic associations between animals and inherited micro-organisms are widespread in nature. In many cases, hosts may be superinfected with multiple inherited symbionts. Acyrthosiphon pisum (the pea aphid) may harbour more than one facultative symbiont (called secondary symbionts) in addition to the obligate primary symbiont, Buchnera aphidicola. Previously we demonstrated that, in a controlled genetic background, A. pisum infected with either Serratia symbiotica or Hamiltonella defensa (called R- and T-type in that study) were more resistant to attack by the parasitoid Aphidius ervi. Here, we examined the consequences of A. pisum superinfected with both resistance-conferring symbionts. We found that an A. pisum line co-infected with both S. symbiotica and H. defensa symbionts exhibits even greater resistance to parasitism by A. ervi than either of the singly infected lines. Despite this added benefit to resistance, superinfections of S. symbiotica and H. defensa symbionts appeared rare in our survey of Utah A. pisum symbionts, which is probably attributable to severe fecundity costs. Quantitative polymerase chain reaction estimates indicate that while the density of H. defensa is similar in singly and superinfected hosts, S. symbiotica densities increased dramatically in superinfected hosts. Over-proliferation of symbionts or antagonistic interactions between symbionts may be harmful to the aphid host. Our results indicate that in addition to host-symbiont interactions, interactions among the symbionts themselves probably play a critical role in determining the distributions of symbionts in natural populations. PMID- 16720404 TI - Variation in preference for a male ornament is positively associated with female eyespan in the stalk-eyed fly Diasemopsis meigenii. AB - There is currently much interest in mate preferences for sexual ornaments. However, few studies have focused on individual variation in mate preference despite its importance for the rate and direction of sexual selection. Females of the sexually dimorphic stalk-eyed fly, Diasemopsis meigenii, exhibit an unambiguous rejection response towards unattractive males bearing small ornaments. We investigated individual mate preferences using repeated sequential sampling of female rejection or acceptance responses to a wide range of male ornament phenotypes. We found significant variation in the strength of individual preference. In addition, preference was positively associated with female eyespan, a condition-dependent trait putatively linked to visual acuity. PMID- 16720405 TI - Variation among species in proteomic sulphur content is related to environmental conditions. AB - The elemental composition of proteins influences the quantities of different elements required by organisms. Here, we considered variation in the sulphur content of whole proteomes among 19 Archaea, 122 Eubacteria and 10 eukaryotes whose genomes have been fully sequenced. We found that different species vary greatly in the sulphur content of their proteins, and that average sulphur content of proteomes and genome base composition are related. Forces contributing to variation in proteomic sulphur content appear to operate quite uniformly across the proteins of different species. In particular, the sulphur content of orthologous proteins was frequently correlated with mean proteomic sulphur contents. Among prokaryotes, proteomic sulphur content tended to be greater in anaerobes, relative to non-anaerobes. Thermophiles tended to have lower proteomic sulphur content than non-thermophiles, consistent with the thermolability of cysteine and methionine residues. This work suggests that persistent environmental growth conditions can influence the evolution of elemental composition of whole proteomes in a manner that may have important implications for the amount of sulphur used by living organisms to build proteins. It extends previous studies that demonstrated links between transient changes in environmental conditions and the elemental composition of subsets of proteins expressed under these conditions. PMID- 16720406 TI - Fungal plant endosymbionts alter life history and reproductive success of aphid predators. AB - Endosymbionts occur in most plant species and may affect interactions among herbivores and their predators through the production of toxic alkaloids. Here, we ask whether effects of mycotoxins produced by the symbiosis of the fungal endophyte Neotyphodium lolii and the grass Lolium perenne are transmitted to the aphidophagous ladybird Coccinella septempunctata when feeding on cereal aphids Rhopalosiphum padi on infected plants. The larval development of coccinellids was extended, while their survival was reduced when feeding exclusively on aphids from infected plants. Ladybirds developing on aphids from infected plants showed reduced fecundity and impaired reproductive performance. Body size and symmetries of ladybird adults were not affected by the endophytes. Consistently strong, negative effects of endophytes on the lifetime performance of ladybirds indicates that mycotoxins are transmitted along food chains, causing significant damage for top predators. Such cascading effects will influence the population dynamics of aphid predators in the long term and could feedback to the primary plant producers. PMID- 16720407 TI - Multiphase versus single pot granulation process: influence of process and granulation parameters on granules properties. AB - High-shear wet granulation is widely used for the production of pharmaceutical dosage forms. Different equipment is available for high-shear granulation and drying. This review focuses on two main processes for granules production: multiphase consisting of high-shear granulation followed by drying in a separate apparatus, and single pot granulation/drying. At present, formulas are specifically developed with regard to the production equipment, which raises many problems when different industrial manufacturing equipment is used. Indeed, final granules properties are likely to depend on equipment design, process, and formulation parameters. Therefore, a good understanding of these parameters is essential to facilitate equipment changes. The aim of this review is to present the influence of equipment, process, and formulation parameters on granules properties, considering both the granulation and the drying steps of multiphase and single pot processes. PMID- 16720408 TI - Noncross-linked copolymers of dimethylaminoethyl methacrylate and methacrylic acid as oral drug carriers. AB - The purpose of this study was to synthesize new water-soluble ampholytic copolymers consisting of tertiary amine and carboxylic acid pendent groups for oral drug carriers. The polymers were prepared with a 1:1 molar ratio of dimethylaminoethyl methacrylate and methacrylic acid by free radical polymerization. After polymerization, polymer rods were recovered, dissolved (or swollen) in de-ionized water, and freeze-dried before obtaining fine powders. Drug release experiments with various drugs, representing a variety of drug solubility and types of amine, were carried out with compressed tablets (total weight of 600 mg) containing a variety of basic drugs in pH's of 1.5 and 7. Surprisingly, zero-order release kinetics even from a tablet geometry has been obtained with drug loading ranging from 20-50%. Drug release in pH 7 maintains a zero-order rate up to 80-85% release after a slight initial burst, whereas in pH 1.5 one may not find the initial burst and zero-order kinetics is extended up to 90-95% release. Drug release becomes faster in pH 1.5 than pH 7 due to the faster rate of protonation of the tertiary amine in acidic conditions. The release of basic drugs in pH 1.5 is not significantly different even with varying solubility and types of amine (primary, secondary, and tertiary). However, different drug release profiles in pH 7 are observed with different types of amine and solubility. PMID- 16720409 TI - Effect of oil phase lipophilicity on in vitro drug release from o/w microemulsions with low surfactant content. AB - In this work we investigated the effects of oil phase lipophilicity on in vitro drug release from topical o/w microemulsions (MEs) containing low percentages of emulsifiers. Three different lipids, isopropyl myristate (IPM), isopropyl palmitate (IPP), and isopropyl stearate (IPS), whose lipophilicity increased in the order IPM < IPP 10.0 mIU/L (highest tertile) was associated (P = 0.049) with increased 12-month risk of cardiovascular death or non-fatal myocardial infarction. Moreover, the combination of failed early reperfusion together with late PAPP-A elevation was strongly (7/13 versus 10/49 patients, P = 0.016) associated with adverse outcome. Admission PAPP-A did not correlate with admission C-reactive protein or cardiac troponin I. CONCLUSIONS: PAPP-A is elevated early in STEMI and then declines rapidly, a pattern consistent with release from the ruptured plaque. The variability of PAPP-A kinetics at 48 hours reflects the success of reperfusion. This study also shows that PAPP-A may have prognostic value in STEMI. PMID- 16720437 TI - Metabolic syndrome aggravates the increased endothelial activation and low-grade inflammation in subjects with familial low HDL. AB - BACKGROUND: Inhibition of cytokine-induced expression of adhesion molecules is one of the atheroprotective mechanisms of high-density lipoprotein (HDL). AIM: We investigated whether increased endothelial activation and low-grade inflammation are present in Finnish subjects with familial low HDL, and which factors contribute to the inflammatory parameters. METHOD: High-sensitivity C-reactive protein (hsCRP), soluble intercellular adhesion molecule-1 (sICAM-1), vascular cell adhesion molecule-1 (sVCAM-1), and sE-selectin were measured in 91 subjects with low HDL-cholesterol from 41 low-HDL families and in 112 normolipidemic controls with comparable age- and gender distribution. Presence of the features of the metabolic syndrome (MetS) was recorded. RESULTS: sVCAM-1, sICAM-1, sE selectin, and hsCRP were significantly higher in low-HDL subjects than in the controls (sVCAM-1: 560+/-147 ng/mL versus 496+/-95 ng/mL, P = 0.001; sICAM-1: 247+/-60 ng/mL versus 215+/-47 ng/mL, P<0.001; sE-selectin: 52+/-20 ng/mL versus 44+/-16 ng/mL, P = 0.022; and hsCRP: 1.73+/-2.05 mg/L versus 0.85+/-1.10 mg/L, P<0.001). Low-HDL subjects had increased body mass index (BMI) and waist, and elevated insulin and triglyceride levels. Adhesion molecules and hsCRP increased according to the number of the features of the MetS. CONCLUSIONS: The presence of the MetS in subjects with familial low HDL-cholesterol aggravates the low-grade inflammation and endothelial activation, and ultimately may add to the higher susceptibility for atherosclerotic disease in these individuals. PMID- 16720439 TI - Does the geometrical arrangement of the outer hair cell stereocilia perform a fluid-mechanical function? AB - CONCLUSIONS: Our experiments qualitatively show that the geometrical structure of the inner ear may have the features of a micro-pump. To further substantiate this hypothesis, additional experiments, particularly on in vivo preparations, are needed. OBJECTIVE: To introduce some new ideas about the functional purpose of the geometric arrangement of the outer hair cell stereocilia. Analogies to some recently developed valveless micro-pumps are pointed out. To illustrate these points, comparative experiments were performed using a simplified macro model. METHODS: Specific structures of the organ of Corti were simulated in a partially open, partially closed acrylic tank. This rough approximation allows the visualization of fluid flows that are generated as a result of the relative motions between the tectorial membrane and the reticular lamina. RESULTS: It was shown that the arrangement of the cochlear elements not only forces fluid to flow in a one-way direction, but also generates a fluid stream that flows through the "outlets" between each two V or W formations of stereocilia. These fluid streams are directed towards the inner hair cells. PMID- 16720438 TI - Spiral ligament fibrocytes release chemokines in response to otitis media pathogens. AB - CONCLUSION: Spiral ligament fibrocytes (SLFs) may be involved in the innate immune response of the inner ear by producing chemoattractants for recruiting inflammatory cells such as neutrophils and monocytes. OBJECTIVE: The purpose of this study was to investigate the cellular responses of SLFs when challenged by inflammatory stimuli such as components of otitis media pathogens or proinflammatory cytokines. MATERIALS AND METHODS: To detect released inflammatory cytokines and chemokines, cells were treated for 48 h with whole lysates of nontypable Haemophilus influenzae (NTHi), Streptococcus pneumoniae, or with interleukin 1 alpha (IL-1alpha). The culture medium was then collected and applied to protein arrays. To compare mRNA levels of chemokines, total RNA was extracted after 3 h of treatment with the above agents, and quantitative real time PCR was performed. RESULTS: Protein array analysis showed that in response to NTHi or S. pneumoniae, rat SLFs released monocyte chemotactic protein 1, macrophage inflammatory protein 3 alpha, TNF-alpha, and cytokine-induced neutrophil chemoattractant 2 and 3. Treatment with IL-1alpha, on the other hand, resulted in release of MCP-1 but not the other molecules. Tissue inhibitor of metalloproteinase 1 and vascular endothelial growth factor were released regardless of the inflammatory stimulus used. PMID- 16720440 TI - Effects of hearing level on habitual sniffing in patients with cholesteatoma. AB - CONCLUSIONS: Quality of hearing is one of the key conditions promoting habitual sniffing associated with closing failure of the Eustachian tube and sniffing may stop with deterioration of hearing due to progression of the disease. OBJECTIVE: To examine habitual sniffing associated with closing failure of the Eustachian tube and hearing level in patients with cholesteatoma. PATIENTS AND METHODS: A total of 171 patients with middle ear cholesteatoma treated in Tohoku University Hospital (106 sides in 101 patients, 63 males and 38 females, mean age 43.3 years), and Sendai Ear Surgi Center (70 sides in 70 patients, 37 males and 33 females, mean age 42.7 years) were included. The patients were divided into two groups based on the presence or absence of habitual sniffing, and pre- and postoperative hearing was compared between the two groups. RESULTS: Habitual sniffing was not found in patients with air conduction hearing level worse than 60 dB or air-bone gap larger than 40 dB. Habitual sniffing disappeared in all patients with expanded air-bone gaps in lower frequency regions after surgery. PMID- 16720441 TI - A longitudinal study of electrical stimulation levels and electrode impedance in children using the Clarion cochlear implant. AB - CONCLUSIONS: Electrical stimulation levels and electrode impedance values (EIVs) in children using the Clarion cochlear implant (CI) programmed with CIS strategy stabilized after 3 months of implant use. The data presented here may be useful as a general guideline for the programming of infants and young children and may further be of help for the identification of patients who fall outside the "average" range. OBJECTIVES: The purpose of the present study was to evaluate changes in electrical stimulation levels, i.e. threshold (T) levels, comfortable (M) levels, dynamic range (DR), and EIVs during the first 18 months of implant use, in children using the Clarion CI. MATERIALS AND METHODS: The maps of 18 pre lingual children (mean age at implantation 4.2 years; range 1-8), using the Enhanced Bipolar 1.2 or Bipolar standard electrode with the S-Series speech processor programmed with CIS strategy, were examined at five time points: connection, and 3, 6, 12, and 18 months post-initial stimulation. T levels, M levels, DR and EIVs were analyzed according to four cochlear segments: apical, apical-medial, medial-basal, and basal. RESULTS: During the first 3 months of implant use T levels increased to some extent, whereas M levels and DR increased significantly. From 3 months and through the entire follow-up, T and M levels as well as DR were stable. EIVs of current carrying electrodes decreased significantly from connection to the 3-month visit; thereafter a stabilization of values was evident. Electrical stimulation levels and EIVs did not differ among the cochlear segments during the entire follow-up. PMID- 16720442 TI - Testing utricular function by means of on-axis rotation. AB - CONCLUSIONS: Subjective visual vertical (SVV) estimation during on-axis rotation provides an efficient screening test of utricle function. The survey demonstrates that isolated disorders of peripheral utricular function can occur while SCC function appears normal. OBJECTIVE: The present study aimed to investigate estimation of SVV during constant velocity yaw rotation (with the head held on axis--to enhance any asymmetry between right and left utricular responses), as a useful screening test. MATERIALS AND METHODS: In all, 230 patients were recruited from the dizziness clinic. For each patient, the SVV was estimated (a) while held stationary, and (b) during constant angular velocity (240 degrees/s), with the head centred on-axis. Bithermal caloric testing was also performed in 201 of the patients. RESULTS: Of those patients with normal SVV results during stationary testing, 18.3% were pathological during rotation testing. In those cases with pathological SVV during stationary testing, a significantly greater deviation from the norm was observed during rotation (p<0.001). Of those patients with normal caloric responses, 44.4% showed pathological SVV estimates; this increased to 54.3% for cases with unilateral weakness, and 56.5% for unilateral loss. No clear correlation was found between reports of tilt illusion and pathological SVV, respectively, between rotatory vertigo and pathological caloric responses. PMID- 16720443 TI - Use of PLGA scaffold for mucociliary epithelium transfer in airway reconstruction: a preliminary study. AB - CONCLUSION: A PLGA biodegradable membrane can be used as a scaffold for mucociliary epithelium transfer. OBJECTIVES: The aim of this study was to examine the usefulness of the PLGA membrane as a biodegradable scaffold for mucociliary epithelium transfer in order for it to be used as a substitute for a skin graft for restoring mucosal defects in the airway. METHODS: A PLGA biodegradable membrane was synthesized using the immersion precipitation method, and morphologic characterization was carried out using scanning electron microscopy (SEM). The degradation test was performed by soaking the PLGA membrane in a culture medium, and the morphological changes were observed by SEM. Human nasal basal epithelial (HNBE) cells were cultured on the newly synthesized PLGA membrane, and the morphological changes were analyzed using SEM. The MUC5AC and MUC8 mRNA levels were analyzed by RT-PCR. RESULTS: The PLGA membrane for the mucociliary epithelium transfer was successfully fabricated. It had a 24 mm diameter, a 50 microm thickness, and many pores with diameters of approximately 3 microm. The PLGA membrane began to degrade from 7 days after it was soaked in the culture medium. It rapidly degraded from 3 weeks and severe destruction of the pore structure was noted from 4 to 6 weeks of soaking. The HNBE cells were well differentiated into the mucociliary epithelium on the PLGA membrane both phenotypically and genotypically. PMID- 16720444 TI - Expression of acidic fibroblast growth factor and basic fibroblast growth factor in nasal polyps. AB - CONCLUSION: Up-regulated expression of acidic fibroblast growth factor (aFGF) and basic fibroblast growth factor (bFGF) in nasal polyps may be related to epithelial proliferation and neoangiogenesis associated with nasal polyp growth. BACKGROUND AND OBJECTIVES: aFGF and bFGF are polypeptides that effect mitogenesis, neoangiogenesis and tissue repair. The purpose of this study was to investigate the expression of aFGF and bFGF at both the mRNA and protein levels in nasal polyps. MATERIALS AND METHODS: Tissue samples of nasal polyp and inferior turbinate mucosae were obtained. Reverse transcriptase (RT)-PCR, in situ hybridization, and Western blotting for aFGF and bFGF were performed in nasal polyp and nasal inferior turbinate. RESULTS: RT-PCR showed 50% aFGF and 29% bFGF expression positivity in 14 nasal polyps; however, 13 turbinates revealed no PCR products. aFGF and bFGF expression was localized to some inflammatory cells of nasal polyps by in situ hybridization. Western blotting identified aFGF and bFGF molecular masses of 18 kDa and 24 kDa, respectively, in nasal polyps, and these levels were higher than those observed in nasal turbinates. aFGF and bFGF proteins were also localized in some inflammatory cells of nasal polyps by immunostaining. PMID- 16720445 TI - The efficacy and safety of once-daily mometasone furoate nasal spray in nasal polyposis: a randomized, double-blind, placebo-controlled study. AB - CONCLUSION: In subjects with mild-to-moderate nasal polyposis, treatment with mometasone furoate nasal spray (MFNS) 200 microg once daily (QD) significantly decreases nasal congestion, reduces polyp size, and improves quality of life. OBJECTIVES: To evaluate the efficacy and safety of MFNS, administered QD in the morning, in subjects with mild-to-moderate nasal polyposis. SUBJECTS AND METHODS: This randomized, double-blind, double-dummy, placebo-controlled clinical trial enrolled subjects with mild-to-moderate nasal polyposis at 12 centers in Denmark, Finland, Norway, and Sweden. Inclusion criteria were: age > or = 18 years, a diagnosis of bilateral nasal polyps, and clinically significant nasal congestion. Following a 2-4-week run-in period, subjects were randomized to receive MFNS 200 microg QD or matching placebo for 16 weeks. RESULTS: A total of 298 subjects were randomized to treatment. Of those subjects included in the intent-to-treat efficacy analysis (n = 291), a statistically greater proportion of the MFNS group than the placebo group had improvements in investigator-assessed nasal congestion score between baseline and end point (the primary outcome) (74.3% vs 46.8%; p < 0.001). Significant benefits of MFNS were also seen for secondary end points, including polyp size, sense of smell, peak nasal inspiratory flow, therapeutic improvement, and quality-of-life measures. MFNS was well tolerated, with no unusual or unexpected adverse events. PMID- 16720446 TI - Can anatomical and functional features in the upper airways predict sleep apnea? A population-based study in females. AB - CONCLUSION: The importance of clinical findings in the nose and throat, including fiberoptic endoscopy during the Muller maneuver, in predicting sleep apnea is greater in normal-weight than in overweight women. OBJECTIVES: The aim of this study was to identify clinical features that could predict sleep apnea in women. METHOD: From 6817 women who previously answered a questionnaire concerning snoring habits, 230 women who reported habitual snoring and 170 women from the whole cohort went through a full-night polysomnography. A nose and throat examination including fiber endoscopic evaluation of the upper airways during the Muller maneuver was performed in a random selection of 132 women aged 20-70 years. RESULTS: Sleep apnea was defined as an apnea-hypopnea index of > or = 10. The influence of clinical features on the prevalence of sleep apnea varied between normal-weight and overweight women. A low soft palate, retrognathia, the uvula touching the posterior pharyngeal wall in the supine position, and a 75% or more collapse at the soft palate during the Muller maneuver were all significant predictors of sleep apnea in women with a body mass index (BMI) < 25 kg/m2 but not in overweight women. PMID- 16720447 TI - Adult laryngeal hemangioma CO2 laser excision. A single institution 3-year experience (Vittorio Veneto 2001-2003). AB - CONCLUSIONS: Adult supraglottic hemangiomas can be treated successfully with CO2 laser excision with limited morbidity. Extended laryngeal cavernous hemangiomas involving hypopharynx should be approached with staged CO2 laser surgical procedures. OBJECTIVES: Hemangioma of the adult larynx is an uncommon, benign lesion characterized by thin, friable mucosa overlying the vascular stroma. The optimal surgical approach to these lesions is still controversial because only anecdotal case reports or very limited series are available. We report a 3-year, retrospective, single institution study of the results of CO2 laser treatment of supraglottic hemangiomas in adults. PATIENTS AND METHODS: Six consecutive cases of adult laryngeal hemangioma were treated by the first author with CO2 laser microsurgery alone. RESULTS: In five of six cases, no recurrences have been diagnosed (median follow-up period: 29 months). One case presented limited persistence of disease in the retro-cricoid and arytenoids at 20-month follow-up control. PMID- 16720448 TI - Simultaneous expression of cyclooxygenase-2 and microsomal prostaglandin E synthase in squamous cell carcinoma of the larynx. AB - CONCLUSION: In laryngeal cancer, arachidonic acid may be metabolized to PGE2 via the cooperative actions of COX-2 and mPGES, which are induced in response to various stimuli. The COX-2-mPGES-PGE2 system may induce differentiation of cancer cells and prevent metastasis, thus improving the survival rate. OBJECTIVE: To examine the expression of COX-1, COX-2, and two downstream enzymes--microsomal PGE synthase (mPGES) and PGD synthase (PGDS)--using immunohistochemistry in human laryngeal squamous cell carcinoma (SCC). PATIENTS AND METHODS: Patients with laryngeal carcinoma were referred to the Department of Otolaryngology for treatment. Formalin-fixed, paraffin-embedded laryngeal carcinoma specimens were obtained from 24 patients. Immunohistochemical expression of COX-1, COX-2, mPGES, LPGDS, and HPGDS was investigated in 24 laryngeal carcinoma samples. RESULTS: Among the carcinomas, cytoplasmic immunoreactivity for COX-2 was found in tumor cells in 18 of 24 cases (72%) and that for mPGES in tumor cells in 23 of 24 cases (92%). The localization of mPGES was very similar to that of COX-2. COX-2 in well differentiated SCCs was higher than in poorly/moderately differentiated SCCs. In terms of lymph node metastasis, there was a significant difference in COX-2 expression. PMID- 16720449 TI - Endoglin expression is associated with poor oncologic outcome in oral and oropharyngeal carcinoma. AB - CONCLUSIONS: The present preliminary results suggest that endoglin (CD105) assessed micro-vessel density (MVD) in primary oral and oropharyngeal squamous cell carcinomas (SCCs) may identify patients at risk of disease recurrence or poor oncological outcome after treatment. OBJECTIVES: MVD is an independent prognostic indicator in several human malignancies. Endoglin antibodies have shown a greater specificity for tumour vasculature in comparison with pan endothelial markers. The present explorative study evaluated endoglin expression and its prognostic role in oral and oropharyngeal SCCs. PATIENTS AND METHODS: The study considered 13 consecutive cases of oral and oropharyngeal SCC with lymph node metastases (pN+) and 13 consecutive pN0 cases. CD105-assessed MVD was calculated at 400x magnification. RESULTS: The mean MVDs were 3.6 and 3.1 in pN+ and pN0 groups, respectively (p>0.05). The mean CD105-assessed MVDs were 4.7 in the group with locoregional recurrence and 2.9 in the group without locoregional recurrence or post-treatment diagnosis of distant metastasis (p=0.01). The mean CD105-assessed MVD in primary oral and oropharyngeal SCCs with poor oncological outcome (recurrence of disease or occurrence of distant metastasis) was 4.3. The mean MVD in primary oral and oropharyngeal SCCs with good outcome was 2.9. Statistical analysis showed a significant difference between CD105-assessed MVD in poor and good outcome groups (p=0.02). PMID- 16720451 TI - Intracochlear misdirected implantation of a cochlear implant. AB - This paper discusses a rare complication of an intracochlear misdirection of the electrode of a cochlear implant in a 55-year-old male. The patient received a cochlear implant using the mastoid-saving surgical approach. Intraoperative measurements showed impedance and NRT reactions suggesting a reasonable function of the inner ear and the implant. Postoperatively our patient suffered from passing vertigo. Postoperative CT scans revealed a misdirection implantation of the cochlear implant into the vestibular part of the inner ear. A deformed implant was removed and a reimplantation was successfully performed after enlarging and reshaping the cochleostomy. Our patient now enjoys all the benefits of an optimal functioning cochlear implant. Intracochlear misdirection of the electrode can occur even when intraoperative measurements seem to be normal. When a patient suffers from unexplained vertigo after cochlear implantation with a poor function of the implant a misdirection of the active electrode must be considered. New high resolution peroperative three-dimensional imaging techniques can probably help to avoid such insertion failures. PMID- 16720450 TI - Supracricoid laryngectomies: long-term oncological and functional results. AB - CONCLUSIONS: Our results confirm that supracricoid laryngectomies (SL) are reliable techniques for glottosupraglottic tumors, even for selected T3 and T4 cases, if the indications are correct. These surgical techniques allow a good quality of life with the preservation of the larynx. OBJECTIVE: SL with cricohyoidoepiglottopexy (CHEP) and cricohyoidopexy (CHP) have been popular over the last 20 years for the treatment of the glottic and/or supraglottic squamous cell carcinoma due to demonstrated good oncological and functional results. We report our experience with these techniques, with special focus on long-term oncological and functional results. PATIENTS AND METHODS: We retrospectively reviewed 206 patients who had undergone SL with CHEP or CHP technique between 1987 and 1998 for glottosupraglottic squamous cell carcinoma in our department. The long-term results for 206 patients with T1-T4 laryngeal carcinomas treated with SL are reported: 90.8% CHEP and 9.2% CHP. The mean follow-up was 62 months. RESULTS: Oncological results: the 5-year actuarial disease-free survival was 85%; the 5-year determinate actuarial survival was 88.3%. Functional results: organ preservation rate was 97%. Phonation was assessed according to the GRBAS scale. PMID- 16720452 TI - Hyalinizing clear cell carcinoma arising from the base of the tongue. AB - Hyalinizing clear cell carcinoma is a low-grade indolent and rare salivary gland tumor originally described by Milchgrub et al. in 1994. We herein report a case of this tumor of the base of the tongue. A 66-year-old Japanese woman presented with a large painless mass in the throat. Computed tomography and magnetic resonance imaging revealed a 40x30-mm well-defined ovoid tumor arising from the base of the tongue. She underwent tracheostomy followed by a resection of the tumor via the transmandibular approach combined with a right-sided supra-omohyoid neck dissection. Because the tumor invasion of the surrounding tissue was limited, the surgical defect at the base of the tongue was relatively small, and no reconstructive procedure needed to be performed. The tumor was histopathologically diagnosed as hyalinizing clear cell carcinoma of the minor salivary gland. Her postoperative clinical course was uneventful. No aspiration or difficulty upon deglutition was recognized when she started transoral ingestion on the eighth postoperative day. The patient is currently free from disease 21 months after surgery. The pathology, clinical characteristics, and treatment of hyalinizing clear cell carcinoma are bibliographically reviewed. PMID- 16720454 TI - Laryngeal metastasis from sigmoid colon adenocarcinoma followed by peristomal recurrence. AB - Considering clinical evidence,<2% of laryngeal malignancies are distant metastases. Cutaneous melanomas are the preponderant primaries metastasizing to the larynx, followed by renal cell carcinomas, breast and lung carcinomas. Colonic adenocarcinoma metastases to the larynx are extremely rare: only 13 cases have been described since 1972. We present the first case of sigmoid colon adenocarcinoma metastasis to the larynx in a patient who developed peristomal malignancy recurrence after total laryngectomy. Patients with metastasis of colonic adenocarcinoma to the larynx usually present with disseminated disease: in view of the poor chances of long-term survival of these patients conservative palliative laryngeal treatments should be carried out to improve the quality of life. Curative treatment should be considered only in isolated laryngeal metastasis. PMID- 16720453 TI - Extranodal Rosai-Dorfman disease: involvement of eye, nose and trachea. AB - Rosai-Dorfman disease (RDD) is a rare non-neoplastic histiocytic proliferative disorder characterized by painless lymphadenopathy. Extranodal lesions frequently occur in the head and neck regions. We report the clinical and histological features of extranodal RDD in a 43-year-old man with a previously unreported combination of multiple gross anterior epibulbar nodules in the right eye, submucosal masses of nasal septum and trachea, and no lymphadenopathy during the 12-year follow-up. The patient underwent ophthalmological, otolaryngological and systemic evaluation; gallium 67 scintigraphy; bronchoscopy; ophthalmic ultrasound; head and neck CT scan; biopsies of epibulbar, nasal and tracheal tissues; and septoplasty. Histological specimens showed lymphocytophagocytosis and positive immunoperoxidase staining for S100 protein in foamy histiocytes; both features were typical for RDD. No response to topical or systemic steroids or to radiation therapy was recorded. Removal of nasal septum masses resolved nasal obstruction. The diagnosis of RDD requires histological and, in challenging cases, immunohistological specimens and is difficult--especially with pure extranodal localizations as in our case. RDD should be suspected in cases of subconjunctival mass and/or submucosal nasal and tracheal swellings not responding to systemic steroids. PMID- 16720455 TI - Oral and intralesional steroid therapy in giant cell granuloma. AB - Giant cell granulomas account for 7% of all benign tumors of the jaw. They are commonly seen as asymptomatic masses. A 4-year-old boy with an exophitic, crusty, hemorrhagic mass on the anterolateral region of the right maxilla was referred to our department. Pathological examination was reported as giant cell granuloma. A mixture of prednisolone, lidocaine and adrenaline was injected intralesionally once a week and oral fluocortone was added to this therapy. A limited surgery could be performed. Intralesional prednisolone and systemic fluocortone therapy seems to be effective, as the lesion stopped growing and increase in bony structures was observed radiologically. PMID- 16720456 TI - Sarcomatoid carcinoma of the parotid gland with apparent metastasis of epidermoid elements to cervical lymph nodes. AB - A case of sarcomatoid carcinoma of the parotid gland is described in a 47-year old male. Despite radical surgery and postoperative chemoradiation therapy, a distant metastasis occurred 1 month after treatment. The primary site was composed of well-differentiated squamous cell carcinoma intermingled with spindle shaped malignant mesenchymal cells. Immunohistochemical studies showed that keratin was present in both epithelial cells and spindle cells, whereas vimentin was present in spindle cells and absent in epithelial cells. Two lymph nodes were metastasized by the epidermoid component. The origin of the sarcomatoid component and the differential diagnosis from malignant mixed tumours are discussed. PMID- 16720457 TI - Lysosomal diseases: natural course, pathology and therapy. Proceedings and abstracts of the 5th International Symposium, Valencia, Spain, April 2005. PMID- 16720458 TI - Understanding the biology of X-linked diseases. PMID- 16720459 TI - The pattern of inheritance of X-linked traits is not dominant or recessive, just X-linked. AB - Our modern concepts of genetic inheritance originated nearly a century ago. Early concepts of dominant and recessive inheritance were developed in insects and were subsequently applied to sex-linked inheritance in mammals. Years of clinical experience, however, suggest that the modern-day rules for X-linked dominant and recessive diseases do not explain why so many female carriers of X-linked 'recessive' disorders have an abnormal phenotype. In a review of 32 X-linked diseases we revealed an unexpectedly high degree of intermediate disease penetrance in females that cannot be explained by existing concepts. We recommend that the terms 'dominant' and recessive' be abandoned and that these disorders be referred to as X-linked. In this review we will present modified rules for X linked inheritance and propose hypotheses related to the potential mechanisms that may explain differences in disease expression in females. CONCLUSION: Past assumptions regarding factors that may affect phenotype in heterozygous females do not capture the extraordinarily variable expressivity of X-linked disorders in females and need to be revisited. PMID- 16720460 TI - X-chromosome inactivation: role in skin disease expression. AB - The occurrence of X inactivation in mammals has the consequence that all women are functional mosaics. In X-linked skin disorders, Lyonization usually gives rise to a mosaic pattern, as manifest by the appearance of the lines of Blaschko. This arrangement of lesions is observed in male-lethal X-linked traits, such as incontinentia pigmenti, focal dermal hypoplasia, Conradi-Hunermann-Happle syndrome, oral-facial-digital syndrome type 1 and MIDAS (microphthalmia, dermal aplasia and sclerocornea) syndrome, as well as in various X-linked non-lethal phenotypes, such as hypohidrotic ectodermal dysplasia of Christ-Siemens-Touraine, IFAP (ichthyosis follicularis-alopecia-photophobia) syndrome and X-linked dyskeratosis congenita. Analogous X-inactivation patterns have been documented in human bones, teeth, eyes and, possibly, the brain. Patterns that are distinct from the lines of Blaschko are also seen, such as the lateralization observed in CHILD (congenital hemidysplasia with ichthyosiform nevus and limb defects) syndrome, and the chequerboard pattern seen in women heterozygous for X-linked congenital hypertrichosis. Exceptional cases of either severe or absent involvement in a woman heterozygous for an X-linked trait can be explained by skewing of X inactivation. Some X-linked skin disorders are caused by genes that escape inactivation, which is why heterozygous female 'carriers' of these disorders do not show mosaicism. A well-known example is X-linked recessive ichthyosis due to steroid sulphatase deficiency, the locus for which is situated at the tip of the short arm of the X chromosome and does not undergo Lyonization. On the other hand, in the case of Fabry disease, the gene encoding alpha galactosidase A is subject to inactivation. Remarkably, however, the skin lesions of women do not show a mosaic pattern. CONCLUSION: In the various X-linked skin disorders, affected women show quite dissimilar degrees of involvement and forms of manifestation because X inactivation may give rise to different patterns of functional mosaicism. Paradoxically, no such pattern is observed in women with Fabry disease. Like many X-linked diseases, Fabry disease should neither be called recessive nor dominant, because these dichotomous terms are obscured by the mechanism of X inactivation. PMID- 16720461 TI - Skewed X inactivation in healthy individuals and in different diseases. AB - In female mammalian cells, one of the two X chromosomes is inactivated in early embryonic life. Females are mosaics for two cell populations, one with the maternal and one with the paternal X as the active chromosome. Skewed X inactivation is arbitrarily defined, often as a pattern where 80% or more of the cells show a preferential inactivation of one X chromosome. Inactivation is presumed to be permanent for all descendants of a cell; however, after about 55 years of age, the frequency of skewed X inactivation in peripheral blood cells increases, probably through selection. Unfavourable skewing of X inactivation, where the X chromosome carrying a mutant allele is the predominantly active X, has been found in affected female carriers of several X-linked disorders; however, for many X-linked disorders, a consistent relationship between the pattern of X inactivation and clinical phenotype has been difficult to demonstrate. One reason for this may be that peripheral blood cells are not a representative or relevant tissue in many disorders. In some severe X-linked disorders, post-inactivation selection takes place against the X chromosome carrying the mutant allele, leading to a completely skewed X-inactivation pattern. Skewed X inactivation has also been reported in young females with breast cancer, and may indicate an effect of X-linked genes on the development of this condition. CONCLUSION: The process of X inactivation and the resultant degree of skewing is clearly important for the expression of genetic diseases. It is also important to consider, however, that under normal conditions the frequency of skewed X inactivation increases with age in peripheral blood cells. Analysis of the expression of a large proportion of the genes on the X chromosome has revealed that X-chromosome inactivation is more heterogeneous than previously thought. PMID- 16720462 TI - Disease manifestations and X inactivation in heterozygous females with Fabry disease. AB - AIM: Fabry disease is an X-linked lysosomal storage disorder characterized by an accumulation of neutral glycosphingolipids in multiple organ systems caused by alpha-galactosidase A deficiency due to mutations in the GLA gene. The majority of heterozygous females show the characteristic signs and symptoms of the disease, and some of them are severely affected. The current hypothesis for the occurrence of disease manifestations in females is skewed X inactivation favouring the mutant GLA allele. METHOD: We analyzed the patterns of X inactivation in the leukocytes of 28 biochemically and genetically characterized symptomatic Fabry disease heterozygotes and their correlation with clinical and biochemical disease expression. RESULTS: X inactivation patterns in symptomatic females who are heterozygous for Fabry disease did not differ from those of female controls of the same age (p = 0.669). Thirteen (46%) of the 28 females with Fabry disease showed random X inactivation, ten (36%) moderate skewing, and five (18%) highly skewed X inactivation. Segregation analysis was performed in the families of six females who had highly or moderately skewed X inactivation. In four of these females, skewing favoured the wild-type GLA allele and in the other two skewing favoured the mutant allele. Patterns of X inactivation or the extent of skewing were not related to the severity of clinical manifestations or to residual enzyme activity. CONCLUSION: In this study we provide evidence that heterozygous females with Fabry disease show random X inactivation. Our data do not support the hypothesis that the occurrence and severity of disease manifestations in the majority of Fabry heterozygotes are related to skewed X inactivation. PMID- 16720463 TI - Assessing multiorgan disease severity: scoring systems and applicability in LSDs. PMID- 16720464 TI - The Mainz Severity Score Index (MSSI): development and validation of a system for scoring the signs and symptoms of Fabry disease. AB - Fabry disease is a rare lysosomal storage disorder caused by the deficient activity of the lysosomal enzyme alpha-galactosidase A. The resultant storage of undegraded glycolipids leads to the progressive development of potentially life threatening manifestations affecting multiple organ systems in the body. This paper describes the development of the Mainz Severity Score Index (MSSI), a scoring system for patients with Fabry disease. CONCLUSION: The MSSI score has been proven to be representative in patients with 'classic' Fabry disease and may be useful for monitoring clinical improvement in patients receiving enzyme replacement therapy. PMID- 16720465 TI - The use of scoring systems in patients with haematological malignancy. AB - The utility of predictive systems in patients with haematological malignancy stems both from the heterogeneity of disease presentation and from the inherent disadvantages of therapy. There is an acute morbidity and mortality associated with chemotherapy and bone marrow transplantation, a need for long term follow up, the possibility of treatment failure and the issue of cost. In general, the value of a scoring system is in its ability to quantify disease burden, classify patient subgroups and predict disease course. Successful scoring systems are disease-specific, have been validated in an independent population and are useful in different countries, centres and individuals. There should be good discrimination of poorly responsive and optimally responsive populations and a high rate of prediction of the disease course for individual patients. Optimal utility should be provided by a few variables which are independent, highly predictive and easy to obtain. Other considerations include economic evaluation and adaptation for population changes over time. CONCLUSION: Experience gained from the use of staging and scoring systems in haematology provides useful insights into their potential use in Fabry disease and other lysosomal storage diseases. Such systems may help to predict whom to treat, when to treat and what response is likely to be achieved. PMID- 16720466 TI - CNS pathology and vascular/circulatory abnormalities in Fabry disease. PMID- 16720467 TI - Magnetic resonance imaging changes in Fabry disease. AB - Recognized magnetic resonance imaging (MRI) abnormalities in the brains of patients with Fabry disease include the consequences of infarction and haemorrhage, non-specific white and grey matter lesions, vascular anomalies, in particular dolicho-ectasia, and a characteristic appearance of the posterior thalamus. A preliminary analysis of MRI findings in patients registered in FOS, the Fabry Outcome Survey, indicates that most patients had abnormal scans (25/47). The commonest abnormality, in males and females, was the presence of cerebral white matter lesions, the number of which increased with patient age. CONCLUSION: MRI is a valuable resource for assessing the CNS complications of Fabry disease, and their response to time and treatment. PMID- 16720468 TI - Increased carotid intima-media thickness in the absence of atherosclerotic plaques in an adult population with Fabry disease. AB - AIM: Fabry disease is considered primarily as a progressive small vessel disease, with ischaemic degenerative lesions involving the kidneys, brain and heart. Macrovascular involvement in male patients includes an accelerated wall hypertrophy of the radial artery and a thickening of the intima-media of the common carotid artery. The aim of this study is to evaluate the prevalence and severity of carotid artery atherosclerosis in hemizygous and heterozygous patients with Fabry disease, compared with a matched control population. METHODS: The common carotid artery intima-media thickness (IMT) of 53 patients with Fabry disease (24 men, 29 women) was measured by high-definition ultrasonography, and the presence or absence of atherosclerotic plaques reported. Results were compared with those of 120 age-matched healthy individuals (83 men, 37 women). RESULTS: The common carotid artery IMT was increased to the same extent in male and female patients with Fabry disease (706+/-211 microm and 749+/-395 microm, respectively) compared with that of the control population (614+/-113 microm). In the Fabry population, IMT did not correlate with either systolic blood pressure or with renal function (plasma creatinine). In the control population, only systolic blood pressure was positively and significantly correlated with IMT. Atherosclerotic plaques in the common carotid artery were not observed in any patient with Fabry disease, whereas 34% of the control population had carotid artery plaques, as evidenced by focal non-homogeneous intima-media thickening greater than 1.2 mm. CONCLUSION: This study presents evidence of a major increase in common carotid artery IMT, both in hemizygous and heterozygous patients with Fabry disease, in the absence of focal atherosclerotic plaques. These results suggest that the conduit arteries may be protected from atherosclerosis in Fabry disease. PMID- 16720469 TI - Fabry disease and vascular risk factors: future strategies for patient-based studies and the knockout murine model. AB - Fabry disease is secondary to deficiency of the lysosomal enzyme alpha galactosidase A, leading to altered glycosphingolipid metabolism and accumulation that is often associated with endothelial dysfunction. Current evidence suggests that there is impairment of the vascular nitric oxide pathway, with abnormalities evident in the cerebral circulation and in the dermal vasculature of patients with Fabry disease. Some of these findings have been confirmed in a mouse model of Fabry disease. The murine model, however, allows investigation of Fabry disease at a non-clinical level and a near complete investigation of biological processes within an affected tissue. This is of particular utility in allowing gene expression analysis of clinically inaccessible tissues such as the aorta. CONCLUSION: Future developments in array technology for proteins and DNA single nucleotide polymorphism analysis, together with gene expression microarray analysis, may open a new chapter in our understanding of the biology of lysosomal storage disorders. PMID- 16720470 TI - Damage at the cellular and organ levels in LSDs: possibility for prevention/reversibility with ERT. PMID- 16720471 TI - Lysosomal dysfunction, cellular pathology and clinical symptoms: basic principles. AB - Between 40 and 50 lysosomal storage disorders are known at present. Fine details of the pathogenic process involved are in general not known. This overview highlights the basic principles of lysosomal pathogenesis and the clinical consequences of defective genes involved in lysosomal functions. The subject is discussed in the context of the possibility of prevention and reversal of cellular and organ damage by enzyme replacement therapy. Also presented is a mechanical model for the muscle pathology observed in Pompe disease. Direct mechanical effects of the non-contractile inclusions - glycogen-loaded lysosomes seem to be a key factor in the loss of force during both early and late stages of the disease. CONCLUSION: Each lysosomal storage disorder and each patient with a given lysosomal disorder has unique molecular, pathological and clinical features. But, the order of pathological events is largely the same. Mutations in a gene cause lysosomal dysfunction which, in turn, results in cellular pathology affecting organ structure and function. Clinical symptoms are the ultimate manifestation. The reversibility of symptoms with enzyme replacement therapy will vary according to the disease, as well as the nature and stage of organ pathology. PMID- 16720472 TI - Inhibition and reversibility of renal changes: lessons from diabetic kidney disease. AB - This review describes the clinical characteristics of kidney disease in patients with diabetes in terms of functional and morphological changes, and summarizes the risk factors for progression of disease and the knowledge available today on various treatment modalities. New insights into the pathogenesis of kidney disease in diabetic patients are also reviewed in the context of the nephropathy of Fabry disease. Newly recognized pathways that play a role in the development/progression of kidney disease in patients with diabetes include metabolic factors, (e.g. advanced glycation end products), intracellular signalling proteins (e.g. protein kinase C) and growth factors/cytokines (e.g. growth hormone, insulin-like growth factors, transforming growth factor beta and vascular endothelial growth factor). As classic examples of progress in our understanding of the pathogenesis of kidney disease in patients with diabetes, the relationship between two growth factor/cytokine-systems and the development of diabetic kidney disease is reviewed, including a description of well-known or potential therapeutic strategies targeting the two systems. CONCLUSION: It is hoped that the new pathogenetic insights into diabetic kidney disease may facilitate the development of new drugs for the treatment of this and related kidney diseases. PMID- 16720473 TI - Reversibility of cellular and organ pathology in enzyme replacement trials in animal models of lysosomal storage diseases. AB - Enzyme replacement therapy (ERT) has now become a feasible treatment option for several lysosomal storage diseases (LSDs). Although the rationale behind this approach is straightforward, there are many factors that may influence the efficacy of treatment. The reversibility of cellular and organ pathology depends on several factors including the particular organ targeted, the dose and biodistribution of enzyme, the accessibility of the target cell to the infused enzyme, the abundance of receptors for mannose-6-phosphate and other ligands in the target tissue and the activity of endocytosis. In addition, each lysosomal enzyme is unique and its ability to reverse pathology must be individually determined according to source, glycosylation and phosphorylation status. The extent to which cellular pathology may be corrected depends upon the delivery of sufficient enzyme to the diseased tissues. CONCLUSION: Studies in animal models have identified numerous factors that influence the therapeutic efficacy of ERT. This would suggest that in patients affected by LSDs rigorous evaluation of each therapeutic preparation will be needed. PMID- 16720474 TI - Gaucher disease: multiple lessons from a single gene disorder. AB - Gaucher disease is the most common lysosomal storage disease. It is caused by a deficiency in the lysosomal enzyme glucocerebrosidase, a beta-glucosidase, which results in the accumulation of the lipid glucocerebroside in macrophages throughout the body. Gaucher disease is most common in the Ashkenazi Jewish population, and three mutations of the gene encoding glucocerebrosidase (GBA) have been shown to be prevalent in this population (c.1226 A > C [N370S], 84GG and IVS2[+1]). In non-Jewish patients, the most common mutation is c.1448 G > C (L444P). Until 15 years ago, treatment has been restricted to symptomatic interventions, such as splenectomy or hip replacement. However, there are now specific treatment options - enzyme replacement therapy and substrate reduction therapy. Future developments may include the use of chaperone therapy. CONCLUSION: The lessons that we have learned from Gaucher disease may well be applicable to the development of therapies for some of the other less common lysosomal storage diseases. PMID- 16720481 TI - Hans Spemann (1869-1941): discoverer of the neuronal organizer. PMID- 16720482 TI - Prophylaxis with palivizumab against respiratory syncytial virus infection in infants with congenital heart disease--who should receive it? AB - Respiratory syncytial virus (RSV) infection is the leading cause of infection of the lower respiratory tract in infants, and is also the leading cause of hospitalization during the first year of life. In this issue of Acta Paediatrica, Meberg and Bru report on the incidence and estimated costs of hospitalizations for RSV infections in children with congenital heart disease from a specified region in Norway. CONCLUSION: Incidence figures of hospitalization due to RSV infection in this high-risk group of infants were similar to the results from several other reports. The cost-benefit analysis of prophylaxis against RSV infection in infants with congenital heart disease, using the monoclonal antibody palivizumab, should also take into account potential savings attributable to such prophylaxis. PMID- 16720483 TI - Detecting heart defects in newborn infants--innocent murmurs mixed with silent dangers. AB - Detection of severe heart defects without unnecessary delay is a primary task for providers of infant care. Screening is challenged by the fact that a normal physical examination after birth does not exclude severe defects. Shorter hospital stay for newborn infants and a reduced number of examinations may further hamper a successful mission. Reassuringly, most critical heart defects are reported to be identified early. CONCLUSION: Increased awareness of the limitations of screening and continued monitoring of detection rates, as well as studies on how they can be improved, are warranted. PMID- 16720484 TI - Enhanced intensive care for the neonatal ductus arteriosus. AB - Failure of ductal closure is common in extremely low birth weight infants with significant postnatal morbidities from both pulmonary overcirculation (i.e. chronic lung disease) and/or systemic hypoperfusion (i.e. necrotizing enterocolitis). Early clinical signs of a hemodymanically significant ductus may be non-specific (i.e. hypotension, increasing ventilator requirements, metabolic acidosis) necessitating early screening by echocardiography. Cyclooxygenase inhibitors remain the first-line treatment option. Indomethacin remains the most commonly used agent, despite comparable efficacy and reduced risk of adverse events with ibuprofen. Surgical intervention is recommended after failure of medical therapy, contraindications to medical treatment or fulminating duct related cardiorespiratory deterioration. Wherever possible, surgical intervention in ELBW infants should be avoided in the first week of life due to the potential risks of ischemia-reperfusion cerebral hemorrhage. The postoperative course is often complicated by left ventricular failure, pulmonary edema, and/or hemodynamic instability requiring close monitoring and physiologically relevant therapeutic interventions. PMID- 16720485 TI - Respiratory syncytial virus infections in congenital heart defects- hospitalizations and costs. AB - AIMS: To register hospitalizations for respiratory syncytial virus (RSV) infections and estimate costs of prophylaxis with humanized monoclonal antibodies (palivizumab) against RSV, compared to hospital care, in cases with congenital heart defects (CHDs). METHODS: Population based study with prospective registration of CHDs. Costs for hospital treatment of RSV-infections in CHD patients calculated by means of the Norwegian Diagnosis Related Groups system. RESULTS: In 43 470 infants live born in the population through the 18-year period 1987-2004 a structural CHD was diagnosed in 527 (1.2%). A total of 898 (2.1%) hospitalizations for RSV-infections occurred in the study population 1987-2005. The hospital admittance rate was significantly higher for CHD-cases (4.8%) than for children without CHD (2%) (P = 0.002). Severe CHDs (need for surgery or catheter intervention) had a higher admittance rate (9.2%) compared to the group of remaining CHDs (3.3%) (P = 0.01). Number needed to treat with palivizumab to avoid one hospitalization for RSV-infection in cases of severe CHDs was calculated to 24, at costs of US dollar 195,000. The expenses for palivizuamab prophylaxis in severe CHDs were 31 times that of hospital treatment. CONCLUSION: Prophylaxis with palivizumab in severe CHDs is not cost-effective. PMID- 16720486 TI - Failure to diagnose critical heart malformations in newborns before discharge--an increasing problem? AB - AIM: To determine what proportion of newborns with critical heart defects are discharged undiagnosed from the maternity ward. METHODS: Data were retrieved retrospectively for infants born in our referral area from 1993 to 2001 and undergoing surgical or catheter-based intervention before 2 mo of age because of critical heart defects. RESULTS: 259 full-term infants had critical heart defects. Duct-dependent systemic circulation was present in 129, duct-dependent pulmonary circulation in 106, and 24 infants were not duct dependent but critically ill. In 51 infants (20%) the heart defect was not suspected before discharge from the maternity ward. Such late detection occurred more often in infants with duct-dependent systemic circulation (30%) and in children with defects that were not duct dependent (38%) than in children with duct-dependent pulmonary circulation (4%) (p<0.001). The proportion detected after discharge from the maternity ward increased from 13% in 1993-1995 and 21% in 1996-1998 to 26% in 1999-2001 (p<0.05). CONCLUSION: Many infants with critical congenital heart defects are not diagnosed before discharge from the neonatal maternity ward. The proportion discharged undiagnosed has increased. Neonatal screening fails mainly in children with duct-dependent systemic circulation. PMID- 16720487 TI - Maternal BMI and serum leptin concentration of infants in the first year of life. AB - AIM: To evaluate the relationship between serum leptin concentration in infants and infant and maternal BMI. METHODS: 75 healthy term infants aged 0-12 mo, 51 exclusively breastfed (BF) for at least 4 mo and 24 formula fed (FF), were included in a cross-sectional study. Weight, length and cranial circumference of infants and their mothers were measured, body mass index (BMI) was calculated using the formula weight (kg)/height (m2) and infant serum leptin concentration was determined. RESULTS: A significant positive correlation between infant serum leptin concentration and maternal BMI was observed in BF infants (r = 0.389, p = 0.005), confirmed in a multiple regression model adjusted for infant age and infant BMI (beta = 0.065, p = 0.006), but not in FF infants. As regards the first 4 mo of life, we have already observed a positive correlation between infant serum leptin concentration and maternal BMI in BF infants (beta = 0.113, p = 0.005), but not in FF infants. CONCLUSION: In BF infants, serum leptin concentration correlates positively with maternal BMI. Higher maternal BMI values might increase leptin levels in breast milk, and maternal adiposity could be involved in infant energy balance. PMID- 16720488 TI - Effects of sociodemographic factors on adherence to breastfeeding and other important infant dietary recommendations. AB - AIM: To examine in an affluent and healthy population the association between sociodemographic factors and the adherence to key infant dietary recommendations. METHODS: In a longitudinal prospective study, healthy newborns were selected randomly in maternity wards around Iceland and their diet recorded every month for one year (n = 124). In addition, a 48-hour-weighed-record, including information on added sugar, fruits, vegetables and the use of A and D vitamin drops was completed by 91 (73%) mothers at 9 and 12 months. Information about the mothers' education, smoking, age, parity, and family income was collected (n = 98; 79%). RESULTS: Regression analyses, including sociodemographic factors, showed exclusive breastfeeding to be positively associated with more education (P = 0.022) and non-smoking (P = 0.013) explaining 22% of the variance, and total breastfeeding (exclusive + partial) with non-smoking (P = 0.006). Multiple regression also showed children's intake of added sugar from 9-12 months to be positively associated with mother's smoking (P = 0.022) and negatively with age (P = 0.026), explaining 30% of variance. Sociodemographic factors were associated with children's fruit and vegetable consumption but not with the administration of AD-vitamin drops. However, when tested for significance between groups, younger mothers with fewer children seemed less likely to give AD-vitamin drops. CONCLUSIONS: In a group of mothers getting regular antenatal care and giving birth to healthy infants, sociodemographic factors can predict which mothers need special guidance concerning recommendations about diet in infancy. These were mothers with less education, who smoked, were younger and were having their first or second child. PMID- 16720489 TI - Feeding disorders related to nutrition. AB - AIM: To study the relationship between early feeding disorders and nutritional intake during the second and third year of life. METHODS: 15 children, nine girls and six boys, with a feeding problem identified at the Specialist Child Health Care Unit at a mean age of 14.5 mo were matched with respect to age and sex with healthy control children. Four-day dietary records were collected at inclusion (time 1) and follow-up of the study (time 2). Measures of growth at birth, at times 1 and 2 of the study, and at 3 y of age were obtained from Child Health Centre records. RESULTS: A lower intake of energy, carbohydrate and protein was evident at times 1 and 2 for the group of children with feeding disorders compared to the controls. Zinc and vitamin B6 were the only micronutrients that differed for the feeding disorder group compared to the control group at times 1 and 2. Children with feeding disorders decelerated in weight during the second year of life and in height during the third year of life as compared to controls. CONCLUSION: The criterion of a persistent failure to eat adequately with a subsequent failure to gain weight, required for the diagnosis of feeding disorders in DSM-IV, was supported by the findings in this study. The relation between nutrition and growth in feeding disorders has not, to our knowledge, been reported in previous research. PMID- 16720490 TI - Diarrheal illness in a cohort of children 0-2 years of age in rural Bangladesh: I. Incidence and risk factors. AB - AIM: To describe clinical characteristics and age- and season-specific incidences of diarrheal episodes, and to evaluate risk factors associated with the occurrence of diarrheal disease. METHODS: A total of 252 infants from rural Bangladesh were followed through household surveillance for 2 y from birth during the years 1993-1996. Demographic and household determinants were linked to the probability of illness using logistic regression models. RESULTS: The overall incidence of diarrhea was 4.25 episodes per child per year. Peak rates of overall, acute, and persistent diarrhea occurred in the 6-11-mo and 12-17-mo age groups. Diarrheal rates peaked during the spring and summer. Among host-related characteristics, having a sibling in the household and having had prior diarrhea were significant risk factors for diarrhea. Among environmental characteristics, spring season remained a highly statistically significant risk factor for diarrhea. CONCLUSION: Diarrheal disease continues to be a substantial burden in young children in rural Bangladesh. Most diarrheal episodes are of short duration, and should primarily be treated with oral rehydration therapy to prevent diarrhea-related mortality. Improved knowledge of oral rehydration therapy, feeding during episodes to prevent further malnutrition, prolonged breastfeeding, and the keeping of livestock in corralled areas of the home are advocated. PMID- 16720491 TI - Phagocytosis of Giardia lamblia trophozoites by human colostral leukocytes. AB - AIM: To determine the phagocytic activity of the polymorphonuclear and mononuclear cells present in human colostrum, and to verify the influence of opsonins in the adherence, ingestion and killing of Giardia lamblia trophozoites. METHODS: Polymorphonuclear and mononuclear phagocytes were incubated with G. lamblia trophozoites, in the presence as well as the absence of supernatant of human colostrum (the source of opsonins) for 30, 60 and 120 min. The trophozoites/phagocytes ratio was 1:1, and the percentage of phagocytosed trophozoites was determined by microscopic examination of acridine orange-stained cells. RESULTS: The mononuclear phagocytes presented more functional activity than the polymorphonuclear. The highest indexes of adherence (77.6 +/- 5.1), ingestion (68.9 +/- 5.5) and killing (48.5 +/- 4.9) were obtained through the incubation of mononuclear cells in the presence of colostrum supernatant for 120 min. CONCLUSION: The phagocytes of human colostrum were able to ingest G.lamblia trophozoites and presented microbicidal activity in vitro, suggesting that these phagocytes may act as an additional mechanism of protection against infant giardiasis through breastfeeding. PMID- 16720492 TI - Ten-year trends in obesity among Israeli schoolchildren: 1990-2000. AB - AIM: To analyze trends in childhood body mass index (BMI) in Israel between 1990 and 2000, and to determine the proportion of obese children using US and population-specific reference values. METHODS: Cross-sectional data from 13 284 second- and fifth-grade schoolchildren were collected, including age, sex, height, weight, country of birth, and time since immigration. Age- and sex specific BMI means and centiles were calculated, and the prevalence of obesity was determined using Israeli and US reference values. RESULTS: BMI values at the 95th centile increased monotonously over time in all age and sex categories. Between 1990 and 2000, 95th centile values increased by 12.7% and 11.8% among second-grade boys and girls, respectively. Among fifth-grade children, 95th centile values increased by 10.2% and 8.4%, respectively. Among second graders in 2000, 11.4% of both boys and girls exceeded the BMI value recorded at the 95th centile in 1990. Among fifth-graders in 2000, 10.7% of boys and 11.1% of girls exceeded the 1990 BMI reference value (p for all comparisons < 0.001). The proportion of obese children increased over time using both Israeli and US reference values. CONCLUSION: This substantial increase in childhood obesity poses a serious health threat, and requires implementation of suitable public health interventions. PMID- 16720493 TI - Health-related quality of life in children diagnosed with asthma, diabetes, juvenile chronic arthritis or short stature. AB - AIM: 1) To assess the reliability and validity of the Swedish version of the Child Health Questionnaire (CHQ), 2) to determine the correlation between children's and parents' responses to the CHQ, and 3) to describe and compare responses to the CHQ of four diagnostic groups. METHODS: A total of 199 Swedish children aged 9-16 with diagnoses of asthma (n = 53), diabetes (n = 48), short stature (n = 51) and juvenile chronic arthritis (JCA, n = 47) and their parents answered the CHQ and relevant validation instruments at a clinic check-up. Coefficient alphas were determined for all dimensions of the instrument, and all but four had acceptable to very good reliability (0.75-0.94). RESULTS: Concerning construct validity, the CHQ correlated significantly with appropriate dimensions of the validation instruments. In general, there were significant correlations between the children's and parents' responses. Comparisons between the diagnostic groups showed several significant differences. The short stature group had the highest quality of life and the JCA group the lowest. There were no sex differences, but children who had not reached puberty scored better on the dimensions of mental health and self-esteem. CONCLUSION: The Swedish version of the CHQ is a reliable and valid instrument. Furthermore, it is recommended to ask children themselves about their health-related quality of life. PMID- 16720494 TI - Familial dysautonomia's impact on quality of life in childhood, adolescence, and adulthood. AB - AIM: To evaluate the quality of life (QoL) of children, adolescents, and adults treated for familial dysautonomia (FD), a pervasive neurological disorder. METHODS: The Child Health Questionnaire was completed by parents of 71 patients, while an additional 74 patients completed the Short Form--36. RESULTS: FD imposed a greater physical than psychosocial burden on the child, while the young adults reported both mental and physical quality of life within the average range. Self esteem was problematic and improved with age, while both groups reported lowering physical quality of life as they grew older, with worsening general health that limited their role at school or work. CONCLUSION: Younger FD patients should be closely monitored for lowered self-esteem and referred for counseling when appropriate, while physical and occupational therapy should be provided in advance of expected lowered physical QoL and role fulfillment with increasing age. This becomes important as the need for additional surgical interventions, such as fundoplication with gastrostomy or spinal fusion, contribute to lower physical functioning. Given the high degree of parental involvement required for the varied manifestations of this multisystem disorder, the need for continued parental assessment and psycho-education about this chronic medical illness is warranted. PMID- 16720495 TI - Fever of unknown origin in 185 paediatric patients: a single-centre experience. AB - AIM: We conducted a prospective study to evaluate the causes and outcome in children with fever of unknown origin (FUO). METHODS: From 1990 to 1999, 185 children with FUO were evaluated. Initial evaluation included routine haematological analysis, Epstein-Barr virus (EBV) serology, urine, stool or blood cultures, chest X-ray and tuberculin probe. RESULTS: In 131 (70%) patients diagnosis was established, and 70 (37.8%) had infectious disease. EBV infection was the most common infection followed by visceral leishmaniasis (VL), urinary tract infection (UTI) and tuberculosis. Autoimmune disorders were diagnosed in 24 (12.9%), Kawasaki disease in 12 (6.4%), malignant diseases in 12 (6.4%) and miscellaneous conditions in 15 (8.1%) patients. In the remaining 54 (30%) patients, diagnosis was not established and most of them had self-limited disease. During the investigation, 26 (14%) patients developed serious organ dysfunction and five patients (two with virus-associated haemophagocytic syndrome, one with VL and two unknown) died. CONCLUSION: The most important infectious causes of FUO in our study were EBV infection and VL. Kawasaki disease represented a significant cause of FUO at the beginning of our study because it was not recognized by primary-care physicians. We report myelodysplastic syndrome as another emerging cause of paediatric FUO. Repeated clinical examination and careful use of specific laboratory examinations, invasive diagnostic procedures or imaging are crucial in approaching paediatric FUO. PMID- 16720496 TI - Complications of video-assisted gastrostomy in children with malignancies or neurological diseases. AB - AIM: To test the hypothesis whether the administration of cytostatic drugs close to surgery in children with malignancies influences the rate of postoperative complications. METHOD: Included in the study were 27 children with malignancies and a control group of 27 neurologically impaired children. All the children had nutritional problems and underwent a video-assisted gastrostomy (VAG) operation during the period 1997-2002. The children were postoperatively followed up. All complications were documented according to a protocol by a specially trained nurse and correlated to the time elapsed from completion of the last preoperative or the first postoperative cytostatic drug treatment. The complications in the two groups were compared. RESULTS: The children with malignant diseases did not have more postoperative complications of the VAG than those having neurological defects. There was no correlation to complications regarding timing of the operation and administration of cytostatic drugs. CONCLUSION: This study revealed no aggravated influence of cytostatic drug treatment on early postoperative problems of VAG. The timing of cytostatic drug administration in relation to the surgical intervention did not influence the frequency of postoperative complications. PMID- 16720497 TI - Asthma symptoms in early childhood--what happens then? AB - AIM: To study the outcome in early adulthood for children with early asthma symptoms and to analyse the factors associated with current asthma. METHODS: In a prospective study, we have re-investigated 89/101 children who were hospitalized before the age of two years due to wheezing. The children were investigated using a questionnaire and allergy and bronchial hyper-responsiveness tests at the age of 17-20 years and compared with age-matched controls. RESULTS: In the cohort, 43% had had asthma symptoms in the preceding 12 months compared with 15% in the control group. The strongest risk factors for asthma were current allergy, bronchial hyper-responsiveness and female gender. Female gender and passive smoking in infancy were independent infantile risk factors. In addition to female gender, two pathways led to current asthma: an allergic pathway from family atopy via the development of allergy and another pathway from early passive smoking via hyper-responsiveness and active smoking. CONCLUSION: In children with early wheezing disorder, current allergy, bronchial hyper-responsiveness and female gender were the strongest risk factors for asthma in early adulthood, while female gender and passive smoking in infancy were independent infantile risk factors. The effects of early passive smoking persist longer than previously reported. PMID- 16720498 TI - Increased levels of BAL cysteinyl leukotrienesinacute [corrected] RSV bronchiolitis. AB - BACKGROUND: Cysteinyl leukotrienes (CysLTs), including LTC4, LTD4 and LTE4, are pivotal mediators in the pathophysiology of asthma. AIM: To determine whether CysLT levels are increased in the lower airways of children with respiratory syncytial virus (RSV) bronchiolitis, as they are in asthmatic children, and to investigate a possible heterogeneity in CysLT levels in children with RSV bronchiolitis. METHODS: Bronchoalveolar lavage (BAL) fluids were obtained from children with acute RSV bronchiolitis (n = 20), from children with acute asthma who had no identifiable virus infection (n = 16) and from control subjects (n = 14). BAL cell counts and differentials were determined, and the concentrations of CysLTs were measured by ELISA. RESULTS: CysLT levels in the asthma (70.6 +/- 52.7 pg/ml, p < 0.001) and bronchiolitis groups (21.9 +/- 23.3 pg/ml, p < 0.05) were significantly higher than in the control group (8.7 +/- 5.2 pg/ml). Among bronchiolitis subjects, the eosinophil-positive subgroup (n = 6) showed significantly higher CysLT levels (49.0 +/- 26.7 pg/ml, p = 0.001) than the control group, but this was not observed in the eosinophil-negative subgroup (n = 14, 10.3 +/- 6.3 pg/ml, p = 0.47). CONCLUSION: CysLT levels are increased in the lower airways during RSV bronchiolitis, although their intensities are lower than those in acute asthma. Among bronchiolitis subjects, high CysLT producers could be distinguished from low CysLT producers by the presence of eosinophilia in BAL fluids, suggesting a pathophysiological heterogeneity in RSV bronchiolitis. PMID- 16720499 TI - Hyperglycaemia as a possible marker of invasive fungal infection in preterm neonates. AB - AIM: The incidence of invasive fungal infection in preterm newborns is rising steadily. Early recognition and treatment are imperative, but diagnosis is difficult as data from microbiological investigations are often poor, and clinical and laboratory signs do not help in differentiating bacterial from fungal infections. We evaluated whether glucose intolerance could represent a possible surrogate marker predictor of invasive fungal infection in preterm neonates. METHODS: We performed a case-control study on neonates with birthweight less than 1250 g admitted to our tertiary-level unit during the years 1998-2004 (n = 383), comparing those with invasive fungal infection (n = 45, group A) to matched controls with late-onset sepsis caused by bacterial agents (n = 46, group B). We investigated in both groups the occurrence of hyperglycaemia (serum glycaemia > 215 mg/dl, i.e. 12 mmol/l) in the first month of life, and its temporal relationship with the episodes of sepsis. RESULTS: Hyperglycaemia occurred significantly more often in group A (21/45, 46.6%) than in group B neonates (11/46, 23.9%) (OR 1.95, 95% CI 1.235-4.432, p = 0.008). Moreover, in 19 of 21 (90.4%) neonates with hyperglycaemia in group A, the carbohydrate intolerance episode typically occurred 72 h prior to the onset of invasive fungal infection; in contrast, no temporal relationship was found in neonates with bacterial sepsis (p = 0.002). Correction of hyperglycaemia was successfully achieved in all neonates of both groups, with no significant differences in the number of days of insulin treatment needed to normalize glycaemia (p = 0.15). CONCLUSIONS: Hyperglycaemia is significantly more frequent in neonates who subsequently develop fungal rather than bacterial late-onset sepsis, with a typical 3-d interval. We suggest that a preterm neonate whose birthweight is less than 1250 g in its first month of life should be carefully evaluated for systemic fungal infection whenever signs of carbohydrate intolerance occur. PMID- 16720501 TI - Case 1: otitis, headache and nerve palsy. PMID- 16720502 TI - Case 2: a 4-year old girl with photophobia and somnolence after a head trauma. PMID- 16720503 TI - Asplenia syndrome in a pair of monozygotic twins. AB - Asplenia syndrome is one of the heterotaxy syndromes, which many familial and animal studies suggest are caused by the loss of adequate genetic control of normal left-right asymmetry development. Moreover, there has not been any environmental factor documented to cause these syndromes. Asplenia syndrome occurring in a pair of monozygotic twins is reported. In view of the negative family history, a new germline mutant gene might be the aetiology of our patients. Both twins are associated with some degree of discordant complex heart defects within the context of a high degree of "mirror-image" of the unpaired thoracoabdominal organs. CONCLUSION: This report implies that sporadic asplenia syndrome might associate with new mutations and further genetic study may be indicated. These monozygotic twins' discordant phenotypes imply that some unidentified factors play an important role in their ultimate development of the same genetically determined abnormalities. PMID- 16720504 TI - Endogenous ethanol fermentation in a child with short bowel syndrome. AB - The term "auto-brewery syndrome" has been frequently used to describe patients who show features of alcohol intoxication because of abnormal yeast proliferation after ingesting carbohydrate-rich meals. We present a case of a 3-y-old girl with short bowel syndrome (SBS) who demonstrated signs of alcohol intoxication on repeated occasions. A blood test indicated an ethanol concentration of 15 mmol/l, and cultures from gastric fluid and faeces showed the presence of Candida kefyr. An association was found between the introduction of a carbohydrate-rich fruit drink and the occurrence of symptoms. CONCLUSION: The possibility of endogenous ethanol fermentation should be considered in patients with SBS and the diagnosis of auto-brewery syndrome added to the differential diagnosis list for D-lactic acidosis. Management includes both antifungal treatment and special diet modification. PMID- 16720505 TI - Infantile polyarteritis nodosa presenting as hyponatraemic hypertensive syndrome. AB - The association of arterial hypertension with hyponatraemic dehydration, known as hyponatraemic hypertensive syndrome (HHS), is a rare and serious hypertensive complication. Here, we describe a 17-mo-old girl who presented with severe hyponatraemic dehydration, hypokalaemia, polyuria, and nephrotic-range proteinuria associated with malignant arterial hypertension and systemic inflammatory disease. Diagnosis of classic polyarteritis nodosa (c-PAN) was made on the basis of renal arteriography demonstrating small arterial aneurysms in association with non-aneurismal changes such as arterial cut-off, arterial tapering stenosis and nephrogram perfusion defect. A decrease of blood pressure by antihypertensive treatment resulted in the normalization of HHS abnormalities. However, c-PAN became well controlled only after 4 mo of immunosuppressive therapy. CONCLUSION: The main interest of this case was the uncommon presentation of systemic polyarteritis nodosa in a very young child. Renal ischaemia from intrarenal vessel disease may have been the trigger event for HHS in our case. Management of PAN-associated severe arterial hypertension is based on immunosuppressive and antihypertensive treatment. PMID- 16720506 TI - Brunner's gland hyperplasia in a child with diabetes mellitus and coeliac disease. PMID- 16720508 TI - What is a top scientist like? PMID- 16720507 TI - Seasonality in the incidence of biliary atresia in Japan. PMID- 16720509 TI - Recurrent and transient depressive symptoms around delivery and maternal sensitivity. AB - In this study, the continuity, timing and severity of depressive symptoms around delivery was compared to maternal sensitivity in a community-based sample of 78 first-time mothers. Maternal mood was assessed before delivery (an interview and the General Health Questionnaire), 2 weeks and 4 months after delivery (the Edinburgh Postnatal Depression Scale), and maternal sensitivity 6-8 weeks after delivery (the CARE-Index). Nearly half (46%) of the mothers reported having depressive symptoms at least once, and half of these (23%) reported having such symptoms recurrently. Depressive mothers were significantly lower in sensitivity than non-depressive ones. Both pre- and postnatal, as well as recurrent and transient symptoms, affected the maternal interactive style, and both major and minor depressive symptoms were in most cases part of depressive continuity. Most (76%) mothers who reported depressive symptoms 2 weeks after delivery also reported recurrent symptoms. The most significant finding is considered to be that most (75%) mothers who were assessed to be at the risk level in sensitivity reported depressive symptoms. The public health nurses in the well-baby clinics identified only a few depressive mothers with recurrent symptoms. The conclusion of our study was that depressive symptoms at any time point around delivery constitute a risk of maternal interactive functioning, and identification of these problems is essential for well-timed and focused interventions. PMID- 16720510 TI - Construct validity of a newly developed instrument: profile of occupational engagement in people with schizophrenia, POES. AB - It is highly relevant to estimate to what extent a person with schizophrenia engages in occupations and participates in different life situations in order to understand the determinants of well-being in this group of people. This study aimed at examining the construct validity of the instrument Profile of Occupational Engagement in persons with Schizophrenia, POES. Global Assessment of Functioning, GAF, and a scale measuring Satisfaction with daily occupations and Activity level were chosen as standards against which POES was validated. As hypothesized, moderate associations were found between POES and GAF (0.73), Activity level (0.70), and Satisfaction with daily occupations (0.50). Regarding the separate items of POES, the items that concerned the range of occupations performed had the strongest association with Activity level, and the items that concerned ongoing occupations, i.e. Routines and Extent of performing meaningful occupations, correlated most strongly with Satisfaction with daily occupations. Thus, the strongest associations were found for the expected items. Altogether, POES seems to possess satisfactory construct validity and be a construct in its own right, as indicated by correlations of expected size and direction with the selected instruments and for expected items. PMID- 16720511 TI - Antipsychotic treatment and extrapyramidal symptoms amongst schizophrenic inpatients. AB - Extrapyramidal symptoms (EPS) are seen in 50-75% of patients treated with typical antipsychotics and are a cause of treatment failure in at least 30% of the patients. Using atypical antipsychotics, the EPS incidence is lower, but a low dosage strategy using typical antipsychotics is also known to cause fewer EPS. What conclusions can be drawn for the daily clinical practice? A naturalistic study including all schizophrenic inpatients in a psychiatric ward (n=123) analysed the effects of treatment concerning positive/negative symptoms, EPS, number of days to re-hospitalization and inpatient-days in the year after baseline admittance, using atypical and typical antipsychotics as recommended by the Danish Society of Psychiatry. The incidence of EPS was significantly higher in patients who were treated with typical antipsychotics in relation to atypical antipsychotics (46% vs. 12%, P<0.001). Patients with EPS had significantly more negative symptoms and a poorer level of function at discharge. Nevertheless, no difference regarding re-hospitalization and inpatient-days was found, whether the patient was treated with typical or atypical antipsychotics. However, it is important to underline that patients treated with atypical oral antipsychotic do as well as patients on typical depot antipsychotics. PMID- 16720512 TI - Important components of a short-term family group programme. From the Danish National Multicenter Schizophrenia Project. AB - Clinicians from three psychiatric departments have established family groups as a specific intervention for the relatives of patients with first-episode psychosis. The intervention manual is combining the psychoeducational model with psychodynamic understanding and principles. The aim of this study was to identify the special elements of the programme that were the most important to the relatives. A questionnaire was developed for the participants of the groups in order to establish their satisfaction concerning 1) The actual knowledge received; 2) improvement in ability to cooperate with the therapeutic system and other public institutions; 3) the possibility of sharing thoughts, feelings and problems; and 4) dealing with feelings of guilt and shame and the possibility of altering the relationship with the mentally ill relative. Thirty-five relatives of 26 patients filled in the questionnaire consisting of 15 specific items. More than 95% of the relatives appreciated the gained knowledge about schizophrenia as well as the possibility of sharing thoughts and feelings with others. Two specific elements had the highest rating as important factors for the learning condition: 1) to listen to others with similar problems and 2) the group leaders' attitude towards patients and relatives. PMID- 16720513 TI - The Children's Depression Inventory (CDI) as measure of depression in Swedish adolescents. A normative study. AB - Self-rating scales are an economical and practical aid in the diagnostic work-up. However, normative data from the general population are needed to interpret scores. Four hundred and five adolescents selected to be fairly representative of the general population (both ethnical Swedes and born abroad) filled in a questionnaire containing the Children's Depression Inventory (CDI), the Multidimensional Anxiety Scale for Children (MASC) and background data (including socio-economic status (SES). The CDI was reliable in terms of internal consistency (0.86) with a mean inter-item correlation of 0.18 and item-total score correlations ranging from 0.26 to 0.57. The CDI sub-scales (Negative Mood, Ineffectiveness, Anhedonia and Negative Self-esteem had good internal consistency values slightly above 0.60, except for sub-scale Interpersonal Problems with poor internal consistency (0.36). The 90th and 95th percentiles respectively were defined by scores 15 and 18 and above for boys and by scores 20 and 23 and above for girls. Common correlates of high scores were female gender, broken family but not SES, nor ethnicity. Also, some estimates of the convergent validity of the CDI were found in a Pearson correlation of 0.40 with the MASC total score and in the capacity of the CDI (predictive validity); OR = 1.1 in predicting suicidal ideation. The CDI has some, but as of yet not sufficient indications of being a valid and reliable measure of depression in adolescence and scores can be used to indicate, though not prove the absence or presence of, depression. PMID- 16720514 TI - Adverse drug reactions of antidepressants and antipsychotics: experience, knowledge and attitudes among Norwegian psychiatrists. AB - Efficient prevention of adverse drug reactions (ADRs) requires knowledge about their severity and pharmacological mechanisms and is dependent on reliable data on their frequencies and possible risk factors. The study was conducted to investigate the prescribers' experience and understanding of the ADRs of psychotropic drugs, and their attitude towards reporting these. In a questionnaire, physicians treating adult psychiatric patients were asked which ADRs that they regarded bothersome for some of the most widely used antidepressants and antipsychotics. Questions about the relationship between blockade of drug receptors and ADRs, and about the physicians' personal experience of and attitudes towards reporting of ADRs were also included. In total, 70 of 91 questionnaires (78%) were returned. The mean number of ADRs regarded bothersome ranged from 2.4 to 9.3 for the various drugs/drug classes. Qualified psychiatrists stated a significantly higher number of bothersome ADRs than did the residents. The percentage of physicians associating blockade of a receptor with a specific ADR varied from 76% (histamine receptor blockade and sedation) to 37% (alpha(1)-adrenergic blockade and tachycardia). Thirty-nine per cent of the physicians had never reported an ADR to the Norwegian Medicines Agency. The number of ADRs considered bothersome was relatively high. The pattern of these ADRs generally mirrored the typical ADR profiles of the drugs. The knowledge of the underlying mechanisms of ADRs was more or less incomplete. The reporting rate of ADRs to the national regulatory authorities was low. PMID- 16720515 TI - Video interaction guidance offered to mothers with postnatal depression: experiences from a pilot study. AB - In the period January-August 2003, a pilot study took place connected to a research project where video interaction guidance with postnatal depressed mothers was a central part. We wanted to gain some experiences with video interaction guidance with focus on the baby. The data were collected through interviews and video recordings and analysed in a phenomenological perspective. In short, the experiences from the guidance's with the six mothers can be described as follows: the mothers experienced their own coping, the mothers engaged in mutual dialogue with their babies and the mothers suffering was recognized. Our hypothesis is that the pictures on the TV screen are especially powerful to bring forth a change in the mothers' self-image. PMID- 16720516 TI - Improved detection and pharmacotherapy of major depression from 1989 to 2001 in psychiatric outpatient care in Finland. AB - There have been several reports of non-detection and undertreatment of major depression during the past decades. In this study, we investigated how accurately major depression was assessed, diagnosed and treated according to gold standards, and whether any trend could be found from 1989 to 2001 in Finland. In total, documents of 4447 patients were retrospectively checked at most four times to find the patients fulfilling the study criteria. Finally, 531 patients were included in the study. The major finding of the study was the improved diagnostic assessment of patients with major depression seen in psychiatric settings. However, a systematic and comprehensive approach in asking about and recording subtyping, severity and comorbidity of depression was insufficient. Another major finding of the study was the improved pharmacotherapy of patients with major depression. It seems to be relevant that problems in the quality of care for depression in psychiatric settings are now more likely to be related to suboptimal intensity and monitoring of treatment than to mere lack of treatment. PMID- 16720517 TI - Brief checklists for assessing violence risk among patients discharged from acute psychiatric facilities: a preliminary study. AB - Violence risk assessment instruments are increasingly being used. Their use has, however, mostly been confined to forensic psychiatry for assessing the risk among perpetrators to repeat violent acts. In general psychiatry, very few studies of violence risk among discharged persons from acute inpatient units have been conducted. The available instruments are extensive and time consuming. This study aimed at the construction of a brief checklist. A 33-item scale, the PS (Preliminary Scheme), strongly influenced by the established HCR-20 (Historical, Clinical and Risk Management Assessment Scheme) was developed to undergo logistic regression analysis and possible item reduction. One hundred and ten patients from an acute inpatient unit, scored with PS at discharge, were monitored for violent episodes throughout the following year. Risk assessments and violence registrations were then compared. Of the 110 patients, 29 (26%) had acted violently during the follow-up, with the PS showing a definite association with post-discharge violence. Receiver operating characteristics (ROC) for the instrument yielded an area under the curve (AUC) of 0.71 (P<0.01). Regression analysis indicated that the number of PS items could be strongly reduced without losing predictive validity. Even a four-item checklist showed a higher AUC (0.77) than the PS did with all 33 items. The four items were: 1) Previous violence, 2) Substance use problems, 3) Lack of empathy and 4) Stress. The development of a brief risk assessment instrument with good predictive properties seems possible. Further clinical trials are planned. Ethical aspects of violence prediction must always be considered. PMID- 16720518 TI - Immunomagnetic separation of adult human olfactory neural progenitors. AB - Olfactory neuroepithelium (ONe) has lifelong regenerative capacity owing to the presence of mitotically active progenitors. The accessibility of ONe makes it a unique source of progenitors for cell replacement strategies in the CNS. We have established lines of neurosphere forming cells (NSFCs) from adult postmortem ONe and patients undergoing nasal sinus surgery by endoscopic biopsy. These heterogeneous lines are composed primarily of an immature neuronally restricted and a small glial restricted subpopulation. More homogeneous subpopulations of the NSFCs are essential for detailed study of factors influencing their lineage restriction. Immunomagnetic bead separation using an antibody against tyrosine kinase (Trk) receptors (Trk-pan, which recognizes Trk-A, B, C) resulted in viable, enriched positive and negative subpopulations that could be analyzed immunocytochemically. The positive cells remained positive for the first week after which the number of Trk-pan expressing cells decreased. The negative subpopulation began to express Trk-pan immunoreactivity after five days in vitro. Both subpopulations reverted to the heterogeneous composition after two weeks. Furthermore, most NSFCs were positive for Trk-B, a few for Trk-A, while no reactivity was observed for Trk-C. Because NSFCs produce brain derived neurotrophic factor (BDNF) and express Trk B, the specific receptor for BDNF, it is likely that population dynamics are under a paracrine and/or autocrine regulatory mechanism. Lineage restriction analysis demonstrated that the isolated subpopulation had a restriction potential equivalent to the original heterogeneous population. These studies characterize further the NSFCs and support the future potential therapeutic use of ONe-derived progenitors for CNS injury and neurodegenerative disorders. PMID- 16720519 TI - Clonal analysis of adult human olfactory neurosphere forming cells. AB - Olfactory neuroepithelium (ONe) is unique because it contains progenitor cells capable of mitotic division that replace damaged or lost neurons throughout life. We isolated populations of ONe progenitors from adult cadavers and patients undergoing nasal sinus surgery that were heterogeneous and consisted of neuronal and glial progenitors. Progenitor lines have been obtained from these cultures that continue to divide and form nestin positive neurospheres. In the present study, we used clonal and population analyses to probe the self-renewal and multipotency of the neurosphere forming cells (NSFCs). NSFCs plated at the single cell level produced additional neurospheres; dissociation of these spheres resulted in mitotically active cells that continued to divide and produce spheres as long as they were subcultured. The mitotic activity of clonal NSFCs was assessed using bromodeoxyuridine (BrdU) incorporation. Lineage restriction of the clonal cultures was determined using a variety of antibodies that were characteristic of different levels of neuronal commitment: ss-tubulin isotype III, neural cell adhesion molecule (NCAM) and microtubule associated protein (MAP2), or glial restriction: astrocytes, glial fibrillary acidic protein (GFAP); and oligodendrocytes, galactocerebroside (GalC). Furthermore, nestin expression, a marker indicative of progenitor nature, decreased in defined medium compared to serum-containing medium. Therefore, adult human ONe-derived neural progenitors retain their capacity for self-renewal, can be clonally expanded, and offer multipotent lineage restriction. Therefore, they are a unique source of progenitors for future cell replacement strategies in the treatment of neurotrauma and neurodegenerative diseases. PMID- 16720520 TI - A method for determining identity and relative purity of carmine, carminic acid and aminocarminic acid. AB - Carmine is one of the few dyes currently certified by the Biological Stain Commission that is not assayed for dye content. Existing assay methods are complex and do not differentiate the three cochineal derivatives carmine, carminic acid and aminocarminic acid. The latter dye is relatively new to the food trade as an acid-stable red colorant and may eventually enter the biological stains market. The assay proposed here is a two-step procedure using quantitative spectrophotometric analysis at high pH (12.5-12.6) followed by a qualitative scan of a low pH (1.90-2.10) solution. Carmine is distinct at high pH, and the remaining dyes are easily distinguished at low pH. Four instances of mislabeling are documented from 18 commercial products, but the mislabeled dyes were not certified dyes. Samples from nearly all lots of carmine certified by the Biological Stain Commission from 1920 to 2004 proved to be carmine, but they varied widely in dye content. Batches from 1920 through the 1940s were significantly richer in dye content. Variability has been extreme since 2000, and most of the poorest lots have been submitted since 1990. PMID- 16720521 TI - A polychromatic staining method for epoxy embedded tissue: a new combination of methylene blue and basic fuchsine for light microscopy. AB - A simple and rapid method is described for staining semithin sections of material embedded in epoxy resin for observing tissues prior to transmission electron microscopy. The method is suitable for tissue fixed with a glutaraldehyde formaldehyde mixture and postfixed in osmium tetroxide. No etching or oxidizing procedures are necessary. Sections 0.5-0.8 microm thick are dried onto a slide and stained with either 0.75% methylene blue and 0.25% azure B or 0.5% methylene blue and 0.5% azure II in 0.5% aqueous borax and heated over a flame for 8-10 sec. The slides are rinsed with water, then stained the same way with 0.1% basic fuchsine in 5% aqueous ethanol. Cytoplasm stains blue; nuclei darker blue; collagen, mucus and elastin pink to red; fat and intracellular lipid droplets gray-green. PMID- 16720522 TI - Evolution of the silver and gold stains in neurohistology. AB - Scientific investigations depend on the reliability of the observations that can be made. This reliability is determined in part by the understanding of the techniques and technology used to make the observations. The limitations and the strengths of the methodology and the equipment used must be evaluated thoroughly. The extent to which this is and has been the case for the use of the metal based stains in neuroscience is the subject of this paper. I evaluate the metallic stains used for neuroscience from several perspectives. I review briefly the state of neurohistology prior to its "golden years," 1870-1910. Then I trace the development of the silver based stains used for neurohistology. I wanted to discuss the reasoning used by the originators of the silver based techniques in developing their specific procedures, but discovered that while procedures may be published, the methods and ideas used to arrive at the final procedures are not usually described in published work. PMID- 16720523 TI - Ultrastructural changes of nucleoli in common wheat induced by actinomycin D. AB - Common wheat root tip meristematic cells were treated with low concentrations of actinomycin D (ActD), then stained whole by silver nitrate. We showed by transmission electron microscopy that the typical nucleolar structure did not form, but a granular and fibrillar network was exhibited in the nucleolar region. Our results support a correlation between nucleolar organization/assembly and the activation of RNA Polymerase I transcription. Furthermore, we speculate that the fibrillar network present in the nucleolar region of ActD treated cells may represent the basic skeletal structure required to support the nucleolus. PMID- 16720526 TI - The energetics of walking on sand and grass at various speeds. AB - This study investigated the energetics of walking on sand and grass. Fourteen adult males, participated in the study. Participants had a mean age of 34.6 years old, 72.6 kg in mass and 172.5 cm in stature, who walked at 3, 4, 5, 6 and 7 km per h on sand and grass surfaces. Physiological measures included heart rate, O(2) uptake, CO(2) exhalation, ventilation and relative O(2) uptake using a MetaMax Ergospirometer. Speed was controlled in a methodology similar to the 'Multistage 20-m Shuttle Run Test'. Data were collected during physiological steady rate at each determined speed. A minimum of 2 h rest was enforced between randomized conditions. Results indicate that there was a significant increase (p < 0.01) in all measured physiological indices indicative of energy expenditure when walking on sand compared to grass at 3-7 km per h, with the greatest disparity between the surfaces (ratio = 1.63) in relative O(2) consumption at 5 km per h. PMID- 16720525 TI - Bedside safety rails: assessment of strength requirements and the appropriateness of current designs. AB - Bedside safety rails are used extensively throughout hospitals and residential care environments as a safety device to protect people against the risk of falling out of bed. However, several adverse incidents involving body entrapment have been linked to their use, which have resulted in serious injury to the bed occupants and, in more extreme cases, fatalities. Internationally agreed standards (i.e. European Committee for Standardisation (CEN) standards) for the design and testing of bed rails have recently been developed and are targeted at improved safety requirements (European Committee for Standardisation 19972000). The purpose of the present study was to: 1) determine whether the CEN test procedure for assessing bed rail strength and rigidity requirements is consistent with human force exertions to which bed rails may be exposed; 2) carry out loading tests on bed rails currently in use throughout the UK to determine whether they comply with the current safety recommendations proposed by the CEN standards.A laboratory study was conducted to determine the force exertions of 20 participants performing seven tasks (including activities of both bed occupants and care providers), which were considered representative of the forces to which bed rails could be exposed during normal use. Forces exerted by participants were measured using a force platform and distortions of the bed rails leading to potential entrapment zones were measured using linear displacement transducers. Static load tests were then conducted, in which incremental point loading was applied at the most adverse points on each of nine different bed rails (commonly used in UK nursing and residential care homes) in accordance with the test procedure detailed in the current CEN standards. Maximum static forces exerted by participants were found to range between 250 N and 350 N, which were within the 500 N force tolerance requirements set by the standards. Dynamic tasks involving 'rolling aggressively' against the bed rail produced the highest peak force of 722 N. None of the bed rails included in this study met the requirements of the CEN test procedures, principally due to horizontal lateral displacements being in excess of 50 mm for a 500 N applied force and/or the bed rails' inability to withstand inward horizontal forces of 500 N. Bed rails used with divan beds were often unable to withstand forces greater than 300 N due to turning moments exceeding the bed rail's restraining mechanism. Current CEN standards for assessing the strength and rigidity of bedside safety rails appear to be consistent with the human forces that might arise as a consequence of the normal movement and activities of a bed occupant or care provider. Although compliance with existing standards is not compulsory, findings from the tests carried out on the selection of bed rails in current use throughout the UK highlight the need for improvements in the design and manufacture of bed rails. PMID- 16720527 TI - Can simple anthropometric and physical performance tests track training-induced changes in maximal box-lifting ability? AB - The objective of this research was to determine if tests of lifting strength and anthropometry could be used to assess training-induced changes in box-lift performance. Ninety-two British Army recruits completed a 10-week training programme. Isometric and dynamic lifting strength, body composition and maximal box-lift to 1.45 m and 1.70 m were assessed before and after training. Training significantly increased box-lift performance, dynamic lifting strength and fat free mass, and reduced body fat. Statistically significant regression models (adjusted R(2) of 0.299 and 0.152) were produced that predicted the changes in box-lift performance from changes in body composition and lifting strength. However, the 95% limits of agreement between the observed and predicted change in box-lift performance showed wide margins of error (11-13 kg; 26-29%). Training induced changes in box-lift performance appear to be highly specific, which suggests that both training and testing procedures should also be specific to box lift performance. PMID- 16720528 TI - MAPO index for risk assessment of patient manual handling in hospital wards: a validation study. AB - Manual handling of disabled patients - as regards movement - is one of the major factors affecting acute low back pain of exposed nursing staff. In the absence of quantitative methods assessing this kind of risk, the Research Unit Ergonomics of Posture and Movement of Milan developed in 1997 a risk assessment method called Movement and Assistance of Hospital Patients (MAPO), which is applicable in hospital wards.A first study conducted in 1999 allowed the identification of three levels of MAPO index corresponding with increasing probabilities of being affected by acute low back pain. In accordance with the well-known traffic light model, for MAPO index values between 0 and 1.5 the risk is considered to be absent or negligible. For values between 1.51 and 5.00 the risk is considered to be moderate. For values exceeding 5.00 the risk is considered to be high. In view of the limitations of the previous study, the results needed confirmation and so, in 2000-2001, another cross-sectional study was carried out, which included 191 hospital wards for acute and chronic patients and 2603 exposed subjects. This paper presents the analytical results of the association between the MAPO index and acute low back pain in this new data sample. The agreement between results of the two studies indicates that the MAPO index can be used as a risk index, although with some caution, as detailed in the paper. It can assess the risk exposure level of patient manual handling in wards and can be a useful tool for planning effective preventive actions to reduce the risk of work-related musculoskeletal disorders in health-care workers looking after disabled patients. PMID- 16720529 TI - Evolvable work-centred support systems for command and control: creating systems users can adapt to meet changing demands. AB - Military command and control (C2) organizations are complex socio-technical systems which must constantly adapt to meet changing operational requirements. We describe our experiences in developing a work-centred support system (WCSS) to aid weather forecasting and monitoring in a military airlift C2 organization as an illustrative case. As part of the development process we conducted field observations both before and after introduction of the WCSS in their operations centre. A striking finding was the constant changes that operations personnel faced (changes in goals and priorities, changes in scale of operations, changes in team roles and structure, and changes in information sources and systems). We describe the changes in workplace demands that we observed and the modifications we needed to make to the WCSS in response. For today's fielded systems, it is seldom possible to make changes that are responsive to users' changing requirements in a timely manner. We argue for the need to incorporate facilities that enable users to adapt their systems to the changing requirements of work and point to some promising directions towards evolvable work-centred support systems. PMID- 16720530 TI - The effects of occupational interventions on reduction of musculoskeletal symptoms in the nursing profession. AB - The objective of the review was to gain more insight into the effects of occupational interventions for primary prevention of musculoskeletal symptoms in healthcare workers. The Cochrane Collaboration methodological guidelines for systematic reviews functioned as a starting point. Thirteen studies meeting the inclusion criteria were analysed for methodological quality and effects. Eight outcome effect areas were established and defined as areas in which an effect had been determined in at least two studies. A method based on levels of scientific evidence was then used to synthesize the information available. Strong scientific evidence for the beneficial effect of occupational interventions was found for the outcome effect areas physical discomfort, technical performance of transfers and frequency of manual lifting. Insufficient evidence was found for the effect areas absenteeism due to musculoskeletal problems, musculoskeletal symptoms, fatigue, perceived physical load and knowledge of risk factors at work and ergonomic principles. Training and education combined with an ergonomic intervention were found to be effective. PMID- 16720531 TI - The differential effect of vibrotactile and auditory cues on visual spatial attention. AB - Previous research has shown that the presentation of spatially predictive auditory and vibrotactile warning signals can facilitate driver responses to driving events seen through the windscreen or rearview mirror. The present study investigated whether this facilitation reflects the priming of the appropriate response (i.e. braking vs. accelerating) or an attentional cuing effect (i.e. a perceptual benefit that facilitates subsequent behavioural responding). In the experiments reported here, participants had to discriminate the colour of a number plate (red vs. blue) following the presentation of either spatially predictive vibrotactile (experiment 1) or auditory (experiment 2) warning signals that indicated the likely location (front or back) of the visual target, while simultaneously performing a highly attention-demanding rapid serial visual presentation task. Numberplate discrimination performance was facilitated following the presentation of valid auditory cues, but not following the presentation of equally informative vibrotactile cues. The use of an orthogonal spatial cuing design enabled with us to rule out of a potential response priming account of these data. The results suggest that whilst directional congruency between a warning signal and a target event may be sufficient to facilitate performance due to the priming of the appropriate response, attentional facilitation effects may also require the co-location of the cue and target within the same functional region of space. PMID- 16720534 TI - Recurrent themes and developments in the history of the Ergonomics Society. AB - This paper provides an outline of the main historical developments and recurrent themes that have taken place in the history of the Ergonomics Society (ES) over 50 years. Alongside a chronology of significant events during the period 1949 1999, a set of issues raised by interviews with prominent members of the ES and other historical materials, we focus specifically on seven main areas: the growth and influence of the ES (e.g. internationalization, membership changes); external relations (e.g. press and public relations); publications (e.g. Society journals and key texts); key individuals and institutions; influence on government and other bodies (e.g. participation in committees and inquiries); and image and identity (e.g. changes to the image of the ES over time). We conclude the paper with a summary and discussion of the key developments in the history, as well as the outstanding challenges for the future. PMID- 16720535 TI - Engagement in democratic health reform. PMID- 16720536 TI - Why communication is crucial: meta-analysis of the relationship between safer sexual communication and condom use. AB - The purpose of this study was to quantitatively synthesize the growing literature on the relationship between safer sexual communication (SSC) among sexual partners and condom use, and to systematically examine a number of conceptual and methodological moderators of this relationship. Data from 53 articles published in 27 journals met criteria for the study. Fifty-five independent effect sizes coded from samples totaling N=18,529 were meta-analyzed. Results indicate that the mean sample-size weighted effect size of the SSC-condom use relation was r=.22, and a number of conceptual variables were found to moderate this relationship. Specifically, communication about condom use (r=.25) and sexual history (r=.23) had significantly (p<.05) larger effect sizes than communication about safer sex (r=.18). In addition, SSC measures operationalized differently had significantly (p<.05) different effect sizes. From largest to smallest, these were behavioral format (r=.29), intentional format (r=.18), and self-efficacy format (r=.13). Measures that tried to assess persuasion attempts as compared with informational exchanges were not found to have significantly different effect sizes (p>.05). Further, methodological moderators tended to be unrelated to effect size. Implications for the future study of safer sexual communication as well as the importance of emphasizing communication skills in HIV preventive interventions are discussed. PMID- 16720537 TI - Effects of communicating social comparison information on risk perceptions for colorectal cancer. AB - People typically believe their health risks are lower than those of others (i.e., optimistic bias). We sought to increase perceptions of colorectal cancer (CRC) risk among adults aged 50-75 who were nonadherent to fecal occult screening (FOBT). 160 participants were randomized to receive information about the following: (1) general CRC risk factors (control), (2) general and tailored CRC risk factor feedback (absolute risk group), or (3) absolute CRC risk factor feedback plus CRC feedback as to how their total number of risk factors compared with that of others (absolute plus comparative risk group). Primary outcomes were perceived absolute and comparative risks, attitudinal ambivalence toward FOBT, and screening intentions; the secondary outcome was return of a completed FOBT. Participants who were told that they had more than the average number of risk factors believed their comparative CRC risk was higher than that of controls and of participants informed that they did not have more than the average number of risk factors. Perceived absolute risk did not vary by group. Participants who received social comparison risk factor feedback expressed greater intentions to screen via a FOBT than participants who received absolute risk feedback and controls; they also expressed less ambivalence about FOBT screening than controls. Although not statistically significant, participants informed they were at lower comparative risk had the highest proportion of completing an FOBT than any other group. These results suggest that providing social comparison CRC risk factor feedback can effectively reduce optimistic comparative risk perceptions. Contrary to findings of models of health behavior change, being informed that one does not have more than the average number of CRC risk factors, while resulting in lower evaluations of perceived comparative risk, did not result in higher ambivalence toward and lower intentions to screen using FOBT or the lowest rate of screening. PMID- 16720538 TI - Advertising and obesity: a behavioral perspective. AB - Concern over the levels of obesity observed in Western countries has grown as researchers forecast a rapid growth in the medical care that a progressively more obese population will require. As health workers deal with increased incidences of diabetes and other obesity-related disorders, policymakers have examined the factors contributing to this problem. In particular, advertising that promotes high fat and high sugar products to children has come under increasing scrutiny. Advertisers have rejected claims that advertising contributes to obesity by arguing that it cannot coerce people into purchasing a product, and does not affect primary demand. This reasoning overlooks the role advertising plays in reinforcing and normalising behavior, however, and it assumes that only direct causal links merit regulatory attention. Ehrenberg's "weak" theory suggests advertising will support unhealthy eating behaviors, while the wide range of sales promotions employed will prompt trial and reward continued consumption. This article presents an alternative analysis of how marketing contributes to obesity and uses behavior modification theory to analyse the "fast-food" industry's promotions. We also review the New Zealand government's response to obesity and suggest policy interventions that would foster healthier eating behaviors. PMID- 16720539 TI - Assessment of cultural sensitivity of cancer information in ethnic print media. AB - Ethnic minority populations prefer cancer information that is respectful of their customs and beliefs about health and illness. Community newspapers are an important source of cancer information for ethnic groups. Our purpose is to evaluate the cultural sensitivity of cancer information in mass print media targeting ethnic minority readership. We assessed for cultural sensitivity 27 cancer articles published in English-language ethnic newspapers (Jewish, First Nations, Black/Caribbean, East Indian) in 2000 using the Cultural Sensitivity Assessment Tool (CSAT). We found that the overall average CSAT score of 27 cancer articles was 2.71. (Scores<2.50 were classified as culturally insensitive.) Articles in First Nations newspapers were more culturally sensitive according to the CSAT (X=2.86), followed by articles in Black/Caribbean (X=2.79) and Jewish (X=2.78) papers. Cancer articles from East Indian newspapers had a mean CSAT score of 2.30 and were classified as culturally insensitive. Four articles were considered culturally sensitive but did not mention ethnic populations as intended readers or as high-risk groups for cancer. We found that, using the CSAT measure, overall, cancer articles in ethnic newspapers included in this study were culturally sensitive. Given limitations of this instrument, we recommend an additional checklist for evaluating the cultural sensitivity of printed cancer information. PMID- 16720541 TI - 7th International Symposium on NeuroVirology, 31 May - 3 June 2006, Philadelphia, Pennsylvania, USA. Abstracts. PMID- 16720544 TI - Peer review system: any other choice? PMID- 16720545 TI - Graft-versus-T-cell non-Hodgkin's lymphoma effect. PMID- 16720546 TI - Lymphoma presenting as Sister Mary Joseph's nodule sparing intra-abdominal involvement. PMID- 16720547 TI - An overview of cytidine deaminases. AB - Enzymes that deaminate cytidine to uridine play an important role in a variety of pathways from bacteria to man. Ancestral members of this family were able to deaminate cytidine only in a mononucleotide or nucleoside context. Recently, a family of enzymes has been discovered with the ability to deaminate cytidines on RNA or DNA. The first member of this new family is APOBEC1, which deaminates apolipoprotein B messenger RNA to generate a premature stop codon. APOBEC1 has the conserved active site motif found in Escherichia coli cytidine deaminase. In addition, APOBEC1 has a unique motif containing 2 phenylalanine residues and an insert of 4 amino acid residues across the active site motif. This motif is present in APOBEC family members including activation-induced cytidine deaminase (AID), APOBEC2, and APOBEC3A through APOBEC3G. AID is essential for initiating class-switch recombination, somatic hypermutation, and gene conversion. The APOBEC3 family is unique to primates. APOBEC3G is able to protect cells from human immunodeficiency virus and other viral infections. This function is not unique to APOBEC3G; other APOBEC3 family members also have this ability. Overexpression of enzymes in this family can cause cancer, suggesting that the genes for the APOBEC family of proteins are proto-oncogenes. Recent advances in the understanding of the mechanism of action of this family are summarized in this review. PMID- 16720549 TI - Lentiviral Vif: viral hijacker of the ubiquitin-proteasome system. AB - Since the beginning of time, microorganisms have been devising ways to bypass detection and destruction by our immune system. Therefore, it is no surprise that along with the identification of the cellular antiviral protein APOBEC3G (A3G) has come the recognition of the viral solution to this assault. Here, we review the research that led up to the identification of A3G and the mechanism that the human immunodeficiency virus protein Vif developed to evade A3G's antiviral activities. PMID- 16720548 TI - The dark side of activation-induced cytidine deaminase: relationship with leukemia and beyond. AB - Activation-induced cytidine deaminase (AID) is a unique cellular enzyme that can trigger point mutations and chromosomal translocations, both of which potentially disturb normal cellular metabolism and affect cancer initiation and progression. The involvement of AID in the progression of leukemia has been suggested by multiple groups on the basis of observations of the statistical correlation between AID expression and a poor prognosis of B-cell chronic lymphocytic leukemia. The fact that ectopic expression of AID in mice results in tumors of the lung and T-lymphocytes suggests an oncogenic role for AID. The inducible nature of AID expression indicates that AID might be induced and cause oncogenic mutations, even in epithelial tissues, where AID expression is absent or very weak under normal conditions. If AID can be induced in epithelial cells by inflammatory signals, as from B-lymphocytes, it may be involved in various pathologic conditions, including inflammation-and infection-associated cancers, for which the molecular mechanism is largely unknown, despite the clinical significance of these diseases. PMID- 16720550 TI - APOBEC family proteins: novel antiviral innate immunity. AB - APOBEC3G has been identified as an anti-human immunodeficiency virus type 1 (HIV 1) host factor that belongs to the APOBEC superfamily of cytidine deaminases. It deaminates cytidine to uridine in nascent minus-strand viral DNA, inducing G-to-A hypermutation in the plus-strand DNA of HIV-1. The accumulating evidence demonstrates that APOBEC family proteins also have an antiviral activity against a wide variety of viruses, including not only retroviruses but also other types of viruses, and that each virus seems to have its own strategy for escaping from APOBEC proteins. These results suggest that the APOBEC3 family plays an important role in antiviral host defense as an innate immunity. Recent progress in research on APOBEC family proteins is reviewed. PMID- 16720551 TI - Genetic risk factors for deep vein thrombosis among Japanese: importance of protein S K196E mutation. AB - There is mounting evidence that mutations associated with a given disease arise with different frequencies among ethnic groups, thus ethnicity-specific studies are needed to identify causative mutations and properly assess risk. In particular, ethnic differences in the genetic background of thrombophilia have been reported. We recently conducted a large-scale analysis of the plasma activities of proteins C, S, antithrombin, and plasminogen within the Japanese general population. We found age- and sex-related differences and estimated the prevalence of deficiencies of protein C (0.13%), antithrombin (0.15%), protein S (1.12%), and plasminogen (4.29%). We also evaluated the genetic contribution to deep vein thrombosis and found that protein S mutation K196E is a genetic risk factor in the Japanese population. We estimated allele frequency to be 0.009, suggesting that 1 of 12,000 Japanese may be homozygous for the E allele, thus possibly as many as 10,000 individuals. Accordingly, a substantial proportion of the Japanese population carries the protein S E allele and is at risk of developing deep vein thrombosis. Given the frequency of this mutation and its strong correlation with deep vein thrombosis, it may be valuable to conduct a large-scale screening for this allele and advise concerned persons to avoid environmental risk factors known to be associated with deep vein thrombosis. PMID- 16720552 TI - Acute painful crises of sickle cell disease in Egyptian children: predictors of severity for a preventive strategy. AB - The objective of this study was to predict which infants with sickle cell disease (SCD) are prone to develop severe painful crises. In a mixed hospital - and community-based population (76 cases), demographic data, SCD diagnostic parameters, and basal blood counts were correlated with 2 indices of SCD severity: pain rate (average number of days of painful episodes per year of follow-up) and serious life-threatening complications, such as hyperhemolytic crises. Data were analyzed blind to these indices. The Student t test, analysis of variance, and Pearson correlation were used to determine association with pain rate. Discriminant analysis was used for the prediction of SCD severity. Pain rate was significantly high in hemoglobin SS patients, especially in those with an early onset of dactylitis. There were statistically significant negative correlations of pain rate with basal hemoglobin level, hematocrit, percent hemoglobin F, and arterial oxygen saturation (P <.01 for all correlations). The top 3 predictors of SCD severity were (in descending order) genotype, basal hemoglobin level, and early dactylitis. Severe forms of SCD could be predicted in early infancy with 100% accuracy by using the basal diagnostic parameters for the disease. These infants should be closely monitored with special attention to ventilation status, even before the development of dactylitis. PMID- 16720553 TI - Molecular mechanism of high hemoglobin F production in Southeast Asian-type hereditary persistence of fetal hemoglobin. AB - Hereditary persistence of fetal hemoglobin (HPFH) is associated with a high level of hemoglobin F (HbF) synthesis in adult heterozygotes. In this study, 2 of 6 unrelated HPFH Thai families were found to be Southeast Asian-type HPFH (SEA HPFH) by analyses of the hematologic data and Southern blot hybridization with polymerase chain reaction-amplified DNA probes. DNA mapping with a probe for a delta-globin fragment showed a 27-kb deletion of DNA that included the beta globin gene and the 3' deoxyribonuclease I hypersensitive site 1 (3'HS1) sequence downstream. Deletion of the insulator, 3'HS1, and the juxta-position of the HPFH 3 core enhancer downstream to the 3' breakpoint have been postulated to be the cause of high HbF production in these individuals. To test this hypothesis, we transfected K562 cells with 4 different bacterial artificial chromosome constructs containing the enhanced green fluorescent protein (EGFP) gene at the position of the Agamma-globin gene (pEBAC/148beta:EGFP). Flow cytometry was used to compare EGFP expression from the pEBAC/148beta:EGFP construct with the HPFH-3 core enhancer immediately 5' to the SEA-HPFH breakpoint (pEnH), from the pEBAC/148beta:EGFP construct with 8 kb of the breakpoint sequence and the HPFH-3 core enhancer (pSEA-HPFH), and from the construct with 3'HS1 followed by the pSEA HPFH sequence (pSEA-HPFH_3pHS1). The results show that high HbF production in SEA HPFH occurs from a deletion of the 3'HS1 sequence and the juxtaposition of the HPFH-3 enhancer downstream to the delta-globin gene. PMID- 16720554 TI - Cyclosporin A as an immunosuppressive treatment modality for patients with refractory autoimmune thrombocytopenic purpura after splenectomy failure. AB - The treatment of autoimmune thrombocytopenic purpura (AITP) remains unsatisfactory in patients refractory to first-line management such as corticosteroid therapy and/or splenectomy. Patients with refractory AITP usually require unacceptably high doses of corticosteroids to maintain a safe platelet count. Immunosuppressive treatment with cyclosporin A (CsA) is a relatively new treatment modality, and no large studies of this drug have been conducted. We used CsA in 6 patients with refractory AITP who had platelet counts of less than 20 x 10(9)/L without any therapy or who had evidence of subcutaneous and mucosal bleeding. All 6 patients had undergone splenectomy. When CsA therapy was begun, 5 of the patients were receiving methylprednisolone (MP) at a daily dose of 32 mg or greater. During the following months, the MP dosage was tapered, or the drug was withdrawn. Three patients achieved a complete remission (CR), whereupon CsA treatment was gradually discontinued. Two of these 3 patients later relapsed, but both responded to an additional course of CsA and achieved a second CR. The remaining 3 patients achieved a partial remission (PR). One patient, a woman with an AITP history of more than 30 years, obtained a stable PR with a platelet count substantially greater than 20 x 10(9)/L, which was successfully maintained by low doses of CsA and MP. The most frequent side effect of CsA therapy in our patients was a painful edema of the lower extremities. Our experience shows that CsA is a safe and effective treatment option for patients with refractory (chronic) AITP. It may be given at a low dose as maintenance therapy, and remissions may be sustained even after the drug has been discontinued. PMID- 16720555 TI - Hypereosinophilia in a patient with invasive thymoma with clonal T-lymphocyte expansion expressing CD4, CD8, and CD25 antigens. AB - We report the case of a patient with hypereosinophilia and invasive thymoma harboring probable clonal proliferation of CD4+, CD8+, and CD25+ T-lymphocytes. A 64-year-old woman had eosinophilia (14.1 x 10(9)/L) and an anterior mediastinal tumor with elevated levels of serum immunoglobulin E (609.8 mg/dL) and interleukin 5 (239 pg/mL). Bone marrow aspirate showed marked infiltration by morphologically normal eosinophils with a normal karyotype but no FIP1L1-PDGFRA fusion gene. Flow cytometric analysis revealed an increasing number of CD3+/CD25+ lymphocytes in the peripheral blood, and the resected thymoma had infiltrated lymphocytes with CD4/CD8/CD25 antigens. Moreover, the thymoma had T-cell receptor rearrangements with a cytogenetically clonal nature, ie, t(2;4)(p22;q26). Although the number of patients with thymoma showing hypereosinophilia is small, this case suggests that a subset of patients with thymoma may have clonal expansion of T-lymphocytes with abnormal phenotypes that affect clinical manifestations, including hypereosinophilia. PMID- 16720556 TI - Therapy-related acute myeloid leukemia 6 years after clonal detection of inv(11)(q21q23) and MLL gene rearrangement. AB - Results of recent studies with animal models suggest that expression of MLL fusion proteins promotes acute leukemogenesis. However, the most potent MLL fusion proteins are not sufficient for the development of acute myeloid leukemia (AML). The clinical data on the pathogenesis of this type of leukemia are limited. We analyzed the case of a patient with therapy-related AML with MLL rearrangement. The patient initially developed AML with t(8;21). Although the patient achieved complete remission with chemotherapy, an abnormal karyotype, inv(11)(q21q23), was detected. After 6-year persistence of a clone with the inversion 11 karyotype in the bone marrow, secondary AML developed. Results of fluorescence in situ hybridization analysis combined with magnet-activated cell sorting analysis showed that MLL rearrangement was detected in CD34+ and CD13+ fractions but not in a CD3+ fraction of the bone marrow. There were 2 important clinical findings. One was that MLL rearrangement was not sufficient for the development of leukemia. The other was that MLL rearrangement targets specific lineages. PMID- 16720557 TI - Bilateral osteonecrosis of the head of the femur during treatment with retinoic acid in a young patient with acute promyelocytic leukemia. AB - All-trans retinoic acid (ATRA) is the drug of choice for the treatment of acute promyelocytic leukemia (APL). In general, ATRA is well tolerated, but it does have side effects, the most severe of which is ATRA syndrome. We report the case of a young patient with APL treated with ATRA for induction and maintenance therapy who then developed avascular necrosis of both femoral heads. We also review cases of APL patients with osteonecrosis of the femoral head after ATRA therapy. PMID- 16720558 TI - Association of plasma adiponectin concentrations with chronic lymphocytic leukemia and myeloproliferative diseases. AB - Adiponectin, an adipocyte-secreted hormone, is an important negative regulator in the immune system and hematopoiesis. In this study, we investigated the association of adiponectin levels with chronic lymphocytic leukemia (CLL) and myeloproliferative diseases (MPDs). We measured adiponectin levels in 19 patients with CLL and 30 patients with MPD (chronic myelogenous leukemia, 15; polycythemia vera, 9; myelofibrosis, 4; essential thrombocythemia, 2). The data were (chronic myelogenous leukemia, 15; polycythemia vera, 9; myelofibrosis, 4; essential thrombocythemia, 2). The data were compared with results from a control group of healthy volunteers who were matched according to age, sex, and body mass index. The adiponectin levels in patients with CLL were lower than in the controls (4.71 +/- 1.33 microg/mL versus 16.61 +/- 3.91 microg/mL; P <.001). They were also significantly lower in patients with MPD than in the controls (8.95 +/- 1.33 microg/mL versus 16.16 +/- 4.77 microg/mL; P <.001). In addition, we compared the adiponectin levels of MPD patients who were treated with interferon (IFN) to the levels of patients who were not treated with IFN. Adipnectin levels were significantly higher in IFN-treated patients (11.03 +/- 1.39 microg/mL versus 6.87 +/- 1.79 microg/mL; P <.001). These results suggest that lymphopoiesis and myelopoiesis negatively influence adiponectin levels. Adiponectin may be related to inflammatory cytokine release. IFN therapy appears to have a positive influence on adiponectin secretion by suppressing inflammatory cytokines. Future studies are needed to prove causality and to provide insight about this hormone's mechanism of action and its potential role regarding the etiology and progression of CLL and MPD. PMID- 16720559 TI - Diffuse pulmonary parenchymal involvement in multiple myeloma: antemortem diagnosis. AB - Antemortem diagnosis of diffuse myelomatous pulmonary infiltrate is rare. We present one such case in which diagnosis was established from bronchoalveolar lavage (BAL) fluid demonstrating the presence of monoclonal plasma cells. The diffuse involvement of the lungs by myeloma was confirmed at autopsy. The possibility of myelomatous pulmonary infiltrates was suggested by the absence of fever or failure to respond to antibiotics but regression with myeloma treatment. Analysis of BAL fluid for the presence of plasma cells and for cytoplasmic immunoglobulin DNA provides a noninvasive means of establishing the diagnosis. Diffuse parenchymal lesions in the lung due to myeloma should be entertained as a differential of pulmonary infiltrates. PMID- 16720560 TI - Bone marrow transplantation with a reduced-intensity conditioning regimen in a patient with Wegener granulomatosis and therapy-related leukemia. AB - We describe a patient with Wegener granulomatosis (WG) who underwent long-term cyclophosphamide treatment and thereafter developed acute myelogenous leukemia (AML). After the AML was induced into remission, the patient received an allogeneic stem cell transplant (allo-SCT) from his sibling after undergoing a reduced-intensity conditioning regimen. His clinical course shortly after allo SCT was uneventful. No clinically apparent acute or chronic graft-versus-host disease developed. Repeated analysis of the peripheral blood lymphocytes after transplantation showed complete donor chimerism. The level of proteinase 3 antineutrophil cytoplasmic antibody (PR3-ANCA) remained undetectable until 4 months after transplantation, when it began to increase. When the level of PR3 ANCA peaked, the patient suddenly presented with fever and joint pain, which later spontaneously resolved in parallel with the declining titer of PR3-ANCA. He is now in remission for both AML and WG at 22 months after transplantation. The patient's clinical course after allo-SCT may provide us with valuable information regarding the establishment of allo-SCT as a therapeutic option for WG. PMID- 16720562 TI - Sinusoidal obstruction syndrome of the liver after hematopoietic stem cell transplantation: decision making for orthotopic liver transplantation. AB - Sinusoidal obstruction syndrome (SOS) is a specific complication of hematopoietic stem cell transplantation (HSCT) that can lead to substantial morbidity and treatment-related mortality. Heparin is frequently used as prophylaxis of and defibrotide as therapy for mild to moderate SOS. In severe cases of SOS these therapies are often ineffective, and orthotopic liver trans-plantation (OLT) may be the only option. Reports in the literature about the outcome of liver transplantation for SOS are contradictory. We describe our second case of OLT after HSCT. The patient died of intracranial hemorrhage 2 weeks after liver transplantation with good initial organ function. In the first case at our center, however, the patient survived more then 8 years. The reported short- to medium-range survival rate for OLT following HSCT is approximately 50%. On the basis of the experience at our center and the findings in a review of the literature, we developed a rational approach to the selection for liver transplantation of patients with life-threatening liver dysfunction after marrow transplantation. PMID- 16720561 TI - Prevention of cytomegalovirus infection by valaciclovir after allogeneic bone marrow transplantation from an unrelated donor. AB - In this prospective single-center study, we evaluated the efficacy and safety of valaciclovir (VACV) in the prevention of cytomegalovirus (CMV) infection after allogeneic bone marrow transplantation (BMT). The study population consisted of 12 patients who underwent allogeneic BMT from an unrelated donor. Patients received acyclovir (ACV) intravenously until they became able to take VACV orally. VACV was administered at a daily dose of 3000 mg and continued until day 100. CMV infection was monitored by CMV antigenemia assay and real-time polymerase chain reaction analysis of plasma. Thirty-five patients who did not receive any form of CMV chemoprophylaxis served as control subjects. CMV infection was detected in 4 (33.3%) of the 12 patients and in 24 (68.6%) of the 35 control subjects (P < .05). The onset of CMV infection was significantly delayed in the VACV group (median, day 43) compared with the control group (median, day 28.5; P < .01). Gastrointestinal symptoms as an adverse event due to VACV administration were observed in 2 patients. The plasma levels of ACV after VACV administration were measured in 8 patients and were similar to those in the healthy subjects. In conclusion, VACV shows normal absorption, even in the early posttransplantation period, and may prevent or delay CMV infection effectively and safely in allogeneic BMT recipients. PMID- 16720563 TI - Intensive immunosuppression therapy for aplastic anemia associated with dyskeratosis congenita. PMID- 16720564 TI - Risk-adapted preemptive therapy for cytomegalovirus disease after allogeneic stem cell transplantation: a single-center experience in Korea. PMID- 16720565 TI - Incidence of and risk factors for severe hepatotoxicity of nelfinavir-containing regimens among HIV-infected patients with chronic hepatitis C. AB - OBJECTIVES: To determine the incidence of and risk factors for severe hepatotoxicity of nelfinavir-containing regimens among human immunodeficiency virus/hepatitis C virus (HIV/HCV)-coinfected patients with known stage of liver fibrosis. METHODS: All HIV/HCV-coinfected patients were monitored for a period of 12 months after starting nelfinavir-containing regimens and, with an available liver biopsy, were included in a retrospective study. RESULTS: A total of 82 patients were included in the study. Nine (10.9%) HIV/HCV-coinfected patients showed an episode of severe hepatotoxicity during the study period. Eight (9.8%) individuals showed grade 3 or 4 change in levels of serum alanine aminotransferase and one subject presented with an event of decompensated liver cirrhosis. Six (18.2%) of 33 patients with advanced liver fibrosis and three (6%) of 49 individuals without advanced liver fibrosis showed an episode of severe hepatotoxicity (P = 0.1). In the multivariate analysis, only nevirapine use during nelfinavir therapy [adjusted odds ratio (AOR) 8.9; 95% confidence interval (CI), 1.4-54.1; P = 0.01] was independently associated with risk of development of severe liver toxicity. CONCLUSIONS: The incidence of severe hepatotoxicity of nelfinavir-containing regimens is low among HIV/HCV-coinfected patients with known stage of liver fibrosis. In addition, our findings show that concomitant nevirapine use is associated with an increased risk of severe hepatotoxicity in these subjects. Likewise, the proportion of severe liver toxicity tended to be higher in individuals with advanced liver fibrosis. PMID- 16720566 TI - Identification of the plasmid-borne quinolone resistance gene qnrS in Salmonella enterica serovar Infantis. AB - OBJECTIVES: A Salmonella enterica serovar Infantis isolate of avian origin was investigated for the presence of the gene qnrS, its transferability and its association with other resistance genes. METHODS: The Salmonella Infantis isolate was investigated for its susceptibility to antimicrobial agents and its plasmid content. Hybridization experiments and PCR assays were performed to identify the resistance genes while transformation and conjugation studies were conducted to show their transferability. The quinolone resistance-determining regions of the genes gyrA, gyrB, parC and parE were sequenced. Moreover, extended sequence analysis was performed to gain insight into the structure and organization of the qnrS gene area. RESULTS: The Salmonella Infantis isolate exhibited the Asp87- >Tyr87 mutation in gyrA, but no resistance-mediating mutations in the other target genes. It carried a conjugative plasmid, pINF5, on which a qnrS gene was detected in close proximity to a Tn3-like, blaTEM-1-carrying transposon. Homology to the qnrS-carrying plasmid pAH0376 of Shigella flexneri was limited to the Tn3 qnrS region. Sequence analysis of an approximately 13.4 kb region of pINF5 identified truncated insertion sequences of types IS26 and IS2 as well as an internal segment of the CS12 fimbrial gene cluster of Escherichia coli up- and downstream of the qnrS gene. CONCLUSIONS: This is to the best of our knowledge the first report of a qnrS gene in a Salmonella isolate. The analysis of the regions flanking the qnrS gene suggested that this region developed in multiple steps that included not only the integration of insertion sequences and a Tn3 like transposon, but also interplasmid recombination events. PMID- 16720567 TI - Extended-spectrum beta-lactamase-producing Enterobacteriaceae in different environments (humans, food, animal farms and sewage). AB - OBJECTIVES: This study aimed to determine the presence of extended-spectrum beta lactamase (ESBL)-producing Enterobacteriaceae in different environments. METHODS: Clinical samples and stool samples from animal farms, sewage, human faecal carriers attending the emergency room and faecal carriers in the context of food borne disease outbreaks were subcultured onto MacConkey agar supplemented with cefotaxime for the detection of ESBL-producing Enterobacteriaceae. Identification, susceptibility pattern and ERIC-PCR were used for clone delineation in each sample. Community consumption of antibiotics was also recorded. RESULTS: An ESBL-producing Enterobacteriaceae prevalence of 1.9% was observed in human infections. A cross-sectional survey of human faecal carriers in the community showed a general prevalence of 6.6% with a temporal distribution. High use of antibiotics in winter coincided with a lower prevalence in carriers. ESBL-producing Enterobacteriaceae were detected in the five samples of human sewage, in samples from 8 of 10 pig farms, 2 of 10 rabbit farms, from all 10 poultry farms and in 3 of 738 food samples studied. Faecal carriage of ESBL-producing Enterobacteriaceae was detected in samples from 19 of 61 food borne outbreaks evaluated. All food-borne outbreaks were due to enteropathogens. The prevalence of carriers in these outbreaks ranged from 4.4% to 66.6%. CONCLUSIONS: This widespread occurrence of ESBL-producing Enterobacteriaceae suggests that the community could act as a reservoir and that food could contribute to the spread of these strains. PMID- 16720568 TI - Chloramphenicol and kanamycin resistance among porcine Escherichia coli in Ontario. AB - OBJECTIVES: The purpose of this study was to compare the distribution of chloramphenicol and kanamycin resistance genes across three populations of porcine Escherichia coli. METHODS: PCR was used to assess the distribution of the major chloramphenicol and kanamycin resistance genes catA1, cmlA and floR, and aphA1, aphA2 and aadB in enterotoxigenic E. coli (ETEC), non-ETEC isolates from cases of diarrhoea and commensal E. coli from healthy pigs. Associations between these genes and resistance genes for other antimicrobials or virulence genes were assessed. RESULTS: The chloramphenicol and kanamycin resistance genes were distributed differently among the three E. coli populations. While aphA1, aphA2 and aadB were evenly distributed among resistant ETEC, non-ETEC and commensals, the catA1 gene was significantly more frequent in ETEC than in non-ETEC and commensals. Transformation experiments confirmed statistical associations by demonstrating that elt, estB, astA, aadA and sul1 were located with catA1 on a large ETEC plasmid. Plasmids carrying cmlA also carried sul3 and aadA. Other plasmids carrying floR and aadB also carried tet(A), sul2, strA/strB, bla(CMY-2) and occasionally aac(3)IV. CONCLUSIONS: The clustering of genes observed is a likely cause for chloramphenicol resistance persistence. Similar to tetracycline, chloramphenicol resistance genes are physically linked to virulence genes. This is not the case for kanamycin resistance determinants, which were linked to other resistance genes only. PMID- 16720569 TI - Does interferon improve portal hypertension? AB - Chronic hepatitis C is the leading cause of advanced liver disease in the Western world. Most patients with liver cirrhosis develop portal hypertension (PHT) (i.e. an abnormal gradient between portal and inferior vena cava pressures), responsible for the most frequent and severe complications of liver disease and, as a consequence, the main cause of death and liver transplantation in those patients. The existence of a potential beneficial effect of antiviral therapy (AVT) on liver inflammation and fibrosis, partially independent of the degree of virological response, has been recently reported. However, the possible influence of these histological changes on PHT has not been evaluated. In this article, we summarize the available findings regarding the effect of AVT on PHT in patients with advanced chronic hepatitis C, as well as its possible implications for clinical practice and future avenues of investigation. PMID- 16720570 TI - Direct evidence that receptor site-4 of sodium channel gating modifiers is not dipped in the phospholipid bilayer of neuronal membranes. AB - In a recent note to Nature, R. MacKinnon has raised the possibility that potassium channel gating modifiers are able to partition in the phospholipid bilayer of neuronal membranes and that by increasing their partial concentration adjacent to their receptor, they affect channel function with apparent high affinity (Lee and MacKinnon (2004) Nature 430, 232-235). This suggestion was adopted by Smith et al. (Smith, J. J., Alphy, S., Seibert, A. L., and Blumenthal, K. M. (2005) J. Biol. Chem. 280, 11127-11133), who analyzed the partitioning of sodium channel modifiers in liposomes. They found that certain modifiers were able to partition in these artificial membranes, and on this basis, they have extrapolated that scorpion beta-toxins interact with their channel receptor in a similar mechanism as that proposed by MacKinnon. Since this hypothesis has actually raised a new conception, we examined it in binding assays using a number of pharmacologically distinct scorpion beta-toxins and insect and mammalian neuronal membrane preparations, as well as by analyzing the rate by which the toxin effect on gating of Drosophila DmNa(v)1 and rat brain rNa(v)1.2a develops. We show that in general, scorpion beta-toxins do not partition in neuronal membranes and that in the case in which a depressant beta-toxin partitions in insect neuronal membranes, this partitioning is unrelated to its interaction with the receptor site and the effect on the gating properties of the sodium channel. These results negate the hypothesis that the high affinity of beta-toxins for sodium channels is gained by their ability to partition in the phospholipid bilayer and clearly indicate that the receptor site for scorpion beta-toxins is accessible to the extracellular solvent. PMID- 16720571 TI - Structure-activity studies on neuropeptide S: identification of the amino acid residues crucial for receptor activation. AB - Neuropeptide S (NPS) has been recently recognized as the endogenous ligand for the previous orphan G-protein-coupled receptor GPR154, now referred to as the NPS receptor (NPSR). The NPS-NPSR receptor system regulates important biological functions such as sleeping/wakening, locomotion, anxiety, and food intake. To collect information on the mechanisms of interaction between NPS and its receptor, a classical structure-activity relationship study was performed. Human (h) NPS derivatives obtained by Ala and d-scan and N- and C-terminal truncation were assessed for their ability to stimulate calcium release in HEK293 cells expressing the human recombinant NPSR. The results of this study indicate that (i) the effect of hNPS is mimicked by the fragment hNPS-(1-10); (ii) Phe(2), Arg(3), and Asn(4) are crucial for biological activity; (iii) the sequence Thr(8) Gly(9)-Met(10) is important for receptor activation, although with non-stringent chemical requirements; and (iv) the sequence Val(6)-Gly(7) acts as a hinge region between the two above-mentioned domains. However, the stimulatory effect of hNPS given intracerebroventricularly on mouse locomotor activity was not fully mimicked by hNPS-(1-10), suggesting that the C-terminal region of the peptide maintains importance for in vivo activity. In conclusion, this study identified the amino acid residues of this peptide most important for receptor activation. PMID- 16720572 TI - The mechanism by which the mitochondrial ATP-sensitive K+ channel opening and H2O2 inhibit the mitochondrial permeability transition. AB - Myocardial infarction is a manifestation of necrotic cell death as a result of opening of the mitochondrial permeability transition (MPT). Receptor-mediated cardioprotection is triggered by an intracellular signaling pathway that includes phosphatidylinositol 3-kinase, endothelial nitric-oxide synthase, guanylyl cyclase, protein kinase G (PKG), and the mitochondrial K(ATP) channel (mitoK(ATP)). In this study, we explored the pathway that links mitoK(ATP) with the MPT. We confirmed previous findings that diazoxide and activators of PKG or protein kinase C (PKC) inhibited MPT opening. We extended these results and showed that other K(+) channel openers as well as the K(+) ionophore valinomycin also inhibited MPT opening and that this inhibition required reactive oxygen species. By using isoform-specific peptides, we found that the effects of K(ATP) channel openers, PKG, or valinomycin were mediated by a PKCepsilon. Activation of PKCepsilon by phorbol 12-myristate 13-acetate or H(2)O(2) resulted in mitoK(ATP) independent inhibition of MPT opening, whereas activation of PKCepsilon by PKG or the specific PKCepsilon agonist psiepsilon receptor for activated C kinase caused mitoK(ATP)-dependent inhibition of MPT opening. Exogenous H(2)O(2) inhibited MPT, because of its activation of PKCepsilon, with an IC(50) of 0.4 (+/-0.1) microm. On the basis of these results, we propose that two different PKCepsilon pools regulate this signaling pathway, one in association with mitoK(ATP) and the other in association with MPT. PMID- 16720573 TI - Ubiquitination and proteasome-dependent degradation of human eukaryotic translation initiation factor 4E. AB - Translation initiation factor 4E (eIF4E) is a cytoplasmic cap-binding protein that is required for cap-dependent translation initiation. Here, we have shown that eIF4E is ubiquitinated primarily at Lys-159 and incubation of cells with a proteasome inhibitor leads to increased eIF4E levels, suggesting the proteasome dependent proteolysis of ubiquitinated eIF4E. Ubiquitinated eIF4E retained its cap binding ability, whereas eIF4E phosphorylation and eIF4G binding were reduced by ubiquitination. The W73A mutant of eIF4E exhibited enhanced ubiquitination/degradation, and 4E-BP overexpression protected eIF4E from ubiquitination/degradation. Because heat shock or the expression of the carboxyl terminus of heat shock cognate protein 70-interacting protein (Chip) dramatically increased eIF4E ubiquitination, Chip may be at least one ubiquitin E3 ligase responsible for eIF4E ubiquitination. PMID- 16720574 TI - Tumor necrosis factor-alpha induces neurotoxicity via glutamate release from hemichannels of activated microglia in an autocrine manner. AB - Glutamate released by activated microglia induces excitoneurotoxicity and may contribute to neuronal damage in neurodegenerative diseases, including Alzheimer disease, Parkinson disease, amyotrophic lateral sclerosis, and multiple sclerosis. In addition, tumor necrosis factor-alpha (TNF-alpha) secreted from activated microglia may elicit neurodegeneration through caspase-dependent cascades and silencing cell survival signals. However, direct neurotoxicity of TNF-alpha is relatively weak, because TNF-alpha also increases production of neuroprotective factors. Accordingly, it is still controversial how TNF-alpha exerts neurotoxicity in neurodegenerative diseases. Here we have shown that TNF alpha is the key cytokine that stimulates extensive microglial glutamate release in an autocrine manner by up-regulating glutaminase to cause excitoneurotoxicity. Further, we have demonstrated that the connexin 32 hemichannel of the gap junction is another main source of glutamate release from microglia besides glutamate transporters. Although pharmacological blockade of glutamate receptors is a promising therapeutic candidate for neurodegenerative diseases, the associated perturbation of physiological glutamate signals has severe adverse side effects. The unique mechanism of microglial glutamate release that we describe here is another potential therapeutic target. We rescued neuronal cell death in vitro by using a glutaminase inhibitor or hemichannel blockers to diminish microglial glutamate release without perturbing the physiological glutamate level. These drugs may give us a new therapeutic strategy against neurodegenerative diseases with minimum adverse side effects. PMID- 16720575 TI - Heparin synergistically enhances interleukin-11 signaling through up-regulation of the MAPK pathway. AB - Using an animal model of heparin-induced osteoporosis we previously demonstrated that heparin causes bone loss, in part, by increasing osteoclast number and activity. Furthermore, we found that, although heparin alone has no effect, it is able to synergistically enhance Interleukin-11 (IL-11)-induced signal transducer and activator of transcription 3 (STAT3) activation and thus increase osteoclast formation in vitro. In the present study, we examine the effect of various serine kinase inhibitors on the ability of heparin to act synergistically with IL-11. Inhibition of the c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (MAPK), or the phosphatidylinositol 3-kinase pathways had no effect on the ability of heparin to promote either IL-11-induced STAT3.DNA complex formation or osteoclast formation in vitro. In contrast, PD098059, a MAPK kinase inhibitor, completely abolished the synergy between heparin and IL-11. In an attempt to resolve the mechanism by which this was occurring, we examined the effect of heparin on STAT3 Ser-727 phosphorylation and extracellular signal-regulated kinases 1 and 2 (Erk1/2) activation, either in the presence or absence of IL-11. Heparin alone was found to have no effect on Ser-727 phosphorylation, nor did heparin alter the phosphorylation status of Ser-727 in the presence of IL-11. Heparin was, however, found to increase Erk1/2 activation in both a time- and dose-dependent manner. When taken together, these findings suggest that heparin enhances IL-11-induced STAT3 activation and thus osteoclast formation, by a mechanism that is independent of STAT3 Ser-727 phosphorylation but that involves up-regulation of the MAPK pathway. PMID- 16720576 TI - Members of RTP and REEP gene families influence functional bitter taste receptor expression. AB - Functional characterization of chemosensory receptors is usually achieved by heterologous expression in mammalian cell lines. However, many chemoreceptor genes, including bitter taste receptors (TAS2Rs), show only marginal cell surface expression. Usually, these problems are circumvented by using chimeric receptors consisting of "export tags" and the receptor sequence itself. It seems likely that chemoreceptor cells express factors for cell surface targeting of native receptor molecules in vivo. For TAS2Rs, however, such factors are still unknown. The present study investigates the influence of RTP and REEP proteins on the functional expression of human TAS2Rs in heterologous cells. We expressed hTAS2Rs in HEK 293T cells and observed dramatic differences in responsiveness to agonist stimulation. By immunocytochemistry we show accumulation of the bitter beta glucopyranoside receptor hTAS2R16 in the Golgi compartment. Coexpression of RTP and REEP proteins changed the responses of some hTAS2Rs upon agonist stimulation, which is likely due to efficient cell surface localization as demonstrated by cell surface biotinylation experiments. The coimmunoprecipitation of hTAS2R16 and RTP3 or RTP4 suggests that the mechanism by which these cofactors influence hTAS2R16 function might involve direct protein-protein interaction. Finally, expression analyses demonstrate RTP and REEP gene expression in human circumvallate papillae and testis, both of which are sites of TAS2R gene expression. PMID- 16720577 TI - Fission yeast Mcm10p contains primase activity. AB - Although Mcm10p is a conserved essential component in eukaryotes required for both the initiation and elongation of DNA chains, its biochemical properties are unknown. Here, we report that the Schizosaccharomyces pombe fission yeast Mcm10 protein contains primase activity. Primases are enzymes that synthesize RNA primers on single-stranded DNA templates that are extended by DNA polymerases. In keeping with this property, Mcm10p supported oligoribonucleotide synthesis of short RNA primers (preferentially initiating synthesis on a dT template) that were extended with dATP by Escherichia coli DNA polymerase I. The C terminus of Mcm10p synthesized RNA, but less efficiently than the full-length protein at low rNTP levels. Mcm10p homologs contain a C-terminal motif found in proteins that polymerize nucleotides. A point mutant within this motif of S. pombe Mcm10p was defective in primer synthesis in vitro, and this mutant failed to support growth in vivo, suggesting that the primase activity of Mcm10p may be essential for cell viability. PMID- 16720578 TI - Evolution of the primate cathelicidin. Correlation between structural variations and antimicrobial activity. AB - Cathelicidin genes homologous to the human CAMP gene, coding for the host defense peptide LL-37, have been sequenced and analyzed in 20 primate species, including Great Apes, hylobatidae, cercopithecidae, callithricidae, and cebidae. The region corresponding to the putative mature antimicrobial peptide is subject to a strong selective pressure for variation, with evidence for positive selection throughout the phylogenetic tree relating the peptides, which favors alterations in the charge while little affecting overall hydrophobicity or amphipathicity. Selected peptides were chemically synthesized and characterized, and two distinct types of behavior were observed. Macaque and leaf-eating monkey RL-37 peptides, like other helical antimicrobial peptides found in insect, frog, and mammalian species, were unstructured in bulk solution and had a potent, salt and medium independent antimicrobial activity in vitro, which may be the principal function also in vivo. Human LL-37 and the orangutan, hylobates, and callithrix homologues instead showed a salt-dependent structuring and likely aggregation in bulk solution that affected antimicrobial activity and its medium dependence. The two types of peptides differ also in their interaction with host cells. The evolution of these peptides has thus resulted in distinct mechanisms of action that affect the direct antimicrobial activity and may also modulate accessory antimicrobial functions due to interactions with host cells. PMID- 16720579 TI - N-linked oligosaccharides on chondroitin 6-sulfotransferase-1 are required for production of the active enzyme, Golgi localization, and sulfotransferase activity toward keratan sulfate. AB - We have shown previously that purified chondroitin 6-sulfotransferase-1 (C6ST-1) was a glycoprotein abundant in N-linked oligosaccharides and could sulfate both chondroitin (C6ST activity) and keratan sulfate (KSST activity); however, functional roles of the N-glycans have remained unclear. In the present study, we show essential roles of N-glycans attached to C6ST-1 in the generation of the active enzyme and in its KSST activity. Treatment with tunicamycin of COS-7 cells transfected with C6ST-1 cDNA totally abolished production of the active C6ST-1. A nearly complete removal of N-glycans of the recombinant C6ST-1 by peptide N glycosidase F increased the C6ST activity but decreased the KSST activity. Among six potential N-glycosylation sites, deletion of the fourth or sixth site from the amino terminus inhibited production of the active C6ST-1, whereas deletion of the fifth site resulted in a marked loss of the KSST activity. Wild-type recombinant C6ST-1 showed a typical Golgi localization, whereas M-4 recombinant C6ST-1, in which the fourth N-glycosylation site was deleted, colocalized with calnexin, an endoplasmic reticulum-resident protein. Unlike wildtype recombinant C6ST-1, M-4 recombinant C6ST-1 showed a weak affinity toward wheat germ agglutinin and was converted completely to the nonglycosylated form by endoglycosidase H. These observations suggest that N-glycan attached to the fourth N-glycosylation site may function in the proper processing of N-glycans required for the Golgi localization, thereby causing the production of the active C6ST-1, and that N-glycan attached to the fifth N-glycosylation site may contribute to the KSST activity of C6ST-1. PMID- 16720580 TI - Candida albicans cell wall ssa proteins bind and facilitate import of salivary histatin 5 required for toxicity. AB - Fungicidal activity of Hst 5 is initiated by binding to cell surface proteins on Candida albicans, followed by intracellular transport to cytoplasmic effectors leading to cell death. As we identified heat shock 70 proteins (Ssa1p and/or Ssa2p) from C. albicans lysates that bind Hst 5, direct interactions between purified recombinant Ssa proteins and Hst 5 were tested by pull-down and yeast two-hybrid assays. Pulldown of both native complexes and those stabilized by cross-linking demonstrated higher affinity of Hst 5 for Ssa2p than for Ssa1p, in agreement with higher levels of interactions between Ssa2p and Hst 5 measured by yeast two-hybrid analyses. C. albicans ssa1Delta and ssa2Delta mutants were constructed to examine Hst 5 binding, translocation, and candidacidal activities. Both ssa1Delta and ssa2Delta mutants were indistinguishable from wild-type cells in growth and hyphal formation. However, C. albicans ssa2Delta mutants were highly resistant to the candidacidal activity of Hst 5, although the ssa1Delta mutant did not have any significant reduction in killing by Hst 5. Total cellular binding of 125I-Hst 5 in the ssa2Delta mutant was reduced to one-third that of wild-type cells, in contrast to the ssa1Delta mutant whose total cellular binding of Hst 5 was similar to the wild-type strain. Intracellular transport of Hst 5 was significantly impaired in the ssa2Delta mutant strain, but only mildly so in the ssa1Delta mutant. Thus, C. albicans Ssa2p facilitates fungicidal activity of Hst 5 in binding and intracellular translocation, whereas Ssa1p appears to have a lesser functional role in Hst 5 toxicity. PMID- 16720582 TI - Financial sustainability planning for immunization services in Cambodia. AB - The expanded programme of immunization was established in Cambodia in 1986. In 2002, 67% of eligible children were immunized, despite significant health sector and macro-economic financial constraints. A financial sustainability planning process for immunization was introduced in 2002, in order to mobilize national and international resources in support of the achievement of child health objectives. The aim of this paper is to outline this process, describe its early impact as an advocacy tool and recommend additional strategies for mobilizing additional resources for health. The methods of financial sustainability planning are described, including the advocacy strategies that were applied. Analysis of financial sustainability planning results indicates rising programme costs associated with new vaccine introduction and new technologies. Despite this, the national programme has demonstrated important early successes in using financial sustainability planning to advocate for increased mobilization of national and international sources of funding for immunization. The national immunization programme nevertheless faces formidable system and financial challenges in the coming years associated with rising costs, potentially diminishing sources of international assistance, and the developing role of sub-national authorities in programme management and financing. PMID- 16720581 TI - Human homologs of Ubc6p ubiquitin-conjugating enzyme and phosphorylation of HsUbc6e in response to endoplasmic reticulum stress. AB - Ubiquitin-conjugating enzyme Ubc6p is a tail-anchored protein that is localized to the cytoplasmic face of the endoplasmic reticulum (ER) membrane and has been implicated in the degradation of many misfolded membrane proteins in yeast. We have undertaken characterization studies of two human homologs, hsUbc6 and hsUbc6e. Both possess tail-anchored protein motifs, display high conservation in their catalytic domains, and are functional ubiquitin-conjugating enzymes as determined by in vitro thiol-ester assay. Both also display induction by the unfolded protein response, a feature of many ER-associated degradation (ERAD) components. Post-translational modification involving phosphorylation of hsUbc6e was observed to be ER-stress-related and dependent on signaling of the PRK-like ER kinase (PERK). The phosphorylation site was mapped to Ser-184, which resides within the uncharacterized region linking the highly conserved catalytic core and the C-terminal transmembrane domain. Phosphorylation of hsUbc6e also did not alter stability, subcellular localization, or interaction with a partner ubiquitin-protein isopeptide ligase. Assays of hsUbc6e(S184D) and hsUbc6e(S184E), which mimic the phosphorylated state, suggest that phosphorylation may reduce capacity for forming ubiquitin-enzyme thiol-esters. The occurrence of two distinct Ubc6p homologs in vertebrates, including one with phosphorylation modification in response to ER stress, emphasizes diversity in function between these Ub-conjugating enzymes during ERAD processes. PMID- 16720583 TI - The burden of psychological symptoms in UK Armed Forces. AB - OBJECTIVE: To assess the prevalence of psychological symptoms during periods of relatively low deployment activity and the factors associated with each psychological health outcome. METHODS: A survey of 4500 randomly selected UK service personnel was carried out in 2002. The questionnaire included the General Health Questionnaire (GHQ-12), the post-traumatic stress disorder checklist (PCL), 15 symptoms and an assessment of alcohol intake. RESULTS: A total of 20% were above cut-offs for GHQ-12, 15% for symptoms, 12% for alcohol intake and 2% for PCL. Gender, age, excessive drinking and smoking were independently associated with most outcomes of interest. Number of deployments was independently associated with multiple symptoms and excessive drinking. High post traumatic stress disorder score was more frequent in the Army and in lower ranks. CONCLUSIONS: Psychological symptoms are highly prevalent in UK Armed Forces. Many risk factors are associated with measures of psychological ill-health. PMID- 16720585 TI - Imprint of evolutionary conservation and protein structure variation on the binding function of protein tyrosine kinases. AB - MOTIVATION: According to the models of divergent molecular evolution, the evolvability of new protein function may depend on the induction of new phenotypic traits by a small number of mutations of the binding site residues. Evolutionary relationships between protein kinases are often employed to infer inhibitor binding profiles from sequence analysis. However, protein kinases binding profiles may display inhibitor selectivity within a given kinase subfamily, while exhibiting cross-activity between kinases that are phylogenetically remote from the prime target. The emerging insights into kinase function and evolution combined with a rapidly growing number of publically available crystal structures of protein kinases complexes have motivated structural bioinformatics analysis of sequence-structure relationships in determining the binding function of protein tyrosine kinases. RESULTS: In silico profiling of Imatinib mesylate and PD-173955 kinase inhibitors with protein tyrosine kinases is conducted on kinome scale by using evolutionary analysis and fingerprinting inhibitor-protein interactions with the panel of all publically available protein tyrosine kinases crystal structures. We have found that sequence plasticity of the binding site residues alone may not be sufficient to enable protein tyrosine kinases to readily evolve novel binding activities with inhibitors. While evolutionary signal derived solely from the tyrosine kinase sequence conservation can not be readily translated into the ligand binding phenotype, the proposed structural bioinformatics analysis can discriminate a functionally relevant kinase binding signal from a simple phylogenetic relationship. The results of this work reveal that protein conformational diversity is intimately linked with sequence plasticity of the binding site residues in achieving functional adaptability of protein kinases towards specific drug binding. This study offers a plausible molecular rationale to the experimental binding profiles of the studied kinase inhibitors and provides a theoretical basis for constructing functionally relevant kinase binding trees. PMID- 16720584 TI - SNPStats: a web tool for the analysis of association studies. AB - SUMMARY: A web-based application has been designed from a genetic epidemiology point of view to analyze association studies. Main capabilities include descriptive analysis, test for Hardy-Weinberg equilibrium and linkage disequilibrium. Analysis of association is based on linear or logistic regression according to the response variable (quantitative or binary disease status, respectively). Analysis of single SNPs: multiple inheritance models (co-dominant, dominant, recessive, over-dominant and log-additive), and analysis of interactions (gene-gene or gene-environment). Analysis of multiple SNPs: haplotype frequency estimation, analysis of association of haplotypes with the response, including analysis of interactions. AVAILABILITY: http://bioinfo.iconcologia.net/SNPstats. Source code for local installation is available under GNU license. PMID- 16720586 TI - Identifying sequence regions undergoing conformational change via predicted continuum secondary structure. AB - MOTIVATION: Conformational flexibility is essential to the function of many proteins, e.g. catalytic activity. To assist efforts in determining and exploring the functional properties of a protein, it is desirable to automatically identify regions that are prone to undergo conformational changes. It was recently shown that a probabilistic predictor of continuum secondary structure is more accurate than categorical predictors for structurally ambivalent sequence regions, suggesting that such models are suited to characterize protein flexibility. RESULTS: We develop a computational method for identifying regions that are prone to conformational change directly from the amino acid sequence. The method uses the entropy of the probabilistic output of an 8-class continuum secondary structure predictor. Results for 171 unique amino acid sequences with well characterized variable structure (identified in the 'Macromolecular movements database') indicate that the method is highly sensitive at identifying flexible protein regions, but false positives remain a problem. The method can be used to explore conformational flexibility of proteins (including hypothetical or synthetic ones) whose structure is yet to be determined experimentally. AVAILABILITY: The predictor, sequence data and supplementary studies are available at http://pprowler.itee.uq.edu.au/sspred/ and are free for academic use. PMID- 16720587 TI - BDT: an easy-to-use front-end application for automation of massive docking tasks and complex docking strategies with AutoDock. AB - MOTIVATION: AutoGrid/AutoDock is one of the most popular software packages for docking, but its automation is not trivial for tasks such as (1) the virtual screening of a library of ligands against a set of possible receptors; (2) the use of receptor flexibility and (3) making a blind-docking experiment with the whole receptor surface. This is an obstacle for research teams in the fields of Chemistry and the Life Sciences who are interested in conducting this kind of experiment but do not have enough programming skills. To overcome these limitations, we have designed BDT, an easy-to-use graphic interface for AutoGrid/AutoDock. AVAILABILITY: BDT is available for free, upon request, for non commercial research. PMID- 16720588 TI - A first look at the new Medicare prescription drug plans. AB - Medicare began offering an outpatient prescription drug benefit through private plans in January 2006. Using nationwide data, we examine the availability, costs, and benefits of regional and local plans offering these benefits to beneficiaries. Because they are an entirely new concept for Medicare, we focus on stand-alone prescription drug plans, comparing them with other Medicare health plans offering drug coverage. The infamous "doughnut hole" exists for all but a few of these plans. One national stand-alone drug plan, though, covers brand-name drugs with no gap. Variations in cost sharing and formularies provide beneficiaries with choices as well as complexity. PMID- 16720589 TI - Effect of short daily home haemodialysis on quality of life, cognitive functioning and the electroencephalogram. AB - BACKGROUND: End-stage renal disease patients have a poor quality of life (QoL), suffer from impaired cognitive functioning, and their electroencephalogram (EEG) shows abnormalities. Conventional haemodialysis (CHD) only partially restores these disorders. Short daily haemodialysis (SDHD) has been reported to improve QoL, but effects on cognitive functioning and EEG have yet to be described. METHODS: Of the 13 patients (11 male, 2 female, age 45.5 +/- 8.1 years), 11 completed the Kidney Disease Quality of Life and Affect Balance Scale questionnaires, 10 underwent neuropsychological testing, and all 13 underwent EEG examination. For the neuropsychological assessments, nine patients (six male, three female, age 45.4 +/- 12.6) who remained on the CHD schedule, served as controls. The dialysis schedule of thrice-a-week for 4 h was changed in the experimental group to six times a week for 2 h (SDHD) over a period of 6 months and back to thrice a week for 4 h. RESULTS: When on SDHD, patients rated several dimensions of health-related QoL as being improved. After resuming CHD, one of these dimensions again decreased and several others worsened even lower than baseline. Cognitive functioning did not change when compared with control data. On the EEG, alpha peak frequency increased slightly when on SDHD but decreased significantly after resuming CHD. CONCLUSIONS: SDHD improves health-related QoL, but has no clear effects on cognitive functioning and EEG. Resumption of CHD after SDHD decreases aspects of QoL and EEG alpha peak frequency but has no effect on cognitive functioning. PMID- 16720590 TI - Back pain in chronic renal failure. PMID- 16720591 TI - A patient with autosomal dominant polycystic kidney and liver disease presenting with urinary tract infection...'Unable to see the wood for the trees'. PMID- 16720592 TI - Does the ID-MS traceable MDRD equation work and is it suitable for use with compensated Jaffe and enzymatic creatinine assays? AB - BACKGROUND: International recommendations suggest that measurement of serum creatinine should be supplemented with an estimate of glomerular filtration rate (GFR) using the Modification of Diet in Renal Disease (MDRD) study equation. One problem has been the lack of standardization of commercially available creatinine assays resulting in varying estimates of GFR. A revision of the MDRD equation offers traceability to a reference method. This study evaluates the use of isotope dilution mass spectrometry (ID-MS), the compensated Jaffe and enzymatic creatinine methods compared with the Beckman CX3 Jaffe assay used to derive the MDRD equation and investigates their impact on GFR estimation using both the original and ID-MS-traceable MDRD equations. METHODS: Serum creatinine was measured in 277 patients by (i) ID-MS, (ii) a Roche enzymatic assay, (iii) a Roche compensated kinetic Jaffe assay and (iv) a Beckman CX3 kinetic Jaffe assay. Estimated GFR was calculated using the MDRD equations. RESULTS: The ID-MS ( 7.5%), Roche enzymatic (-8.6%) and compensated kinetic Jaffe (-11.9%) assays were all negatively biased (P < 0.0001) compared with the Beckman CX3 assay, causing predictable, clinically significant, overestimation of GFR when the original MDRD equation is used. This positive bias was reduced (ID-MS 6.7 to 0.4%; enzymatic 8.8 to 3.4%; compensated kinetic Jaffe 13.7 to 7.1%) when GFRs were calculated using the ID-MS-traceable MDRD equation. CONCLUSIONS: Compensated assays that account for non-creatinine chromogen interference produce significantly higher estimates of GFR when using the original MDRD equation. Use of the ID-MS traceable MDRD equation ameliorates this effect. There is good agreement between estimated GFR derived from the original MDRD equation using Beckman Astra CX3 data and estimated GFR derived from the new ID-MS-traceable MDRD equation using a local ID-MS creatinine assay. This suggests that the ID-MS-traceable MDRD equation may be reliably used with both ID-MS and true ID-MS-traceable creatinine assays without the requirement for standardization to the MDRD laboratory. PMID- 16720593 TI - The effect of statins on urinary albumin excretion and glomerular filtration rate: results from both a randomized clinical trial and an observational cohort study. AB - BACKGROUND: Statins improve cardiovascular outcome, but less is known on the renal outcome. We, therefore, studied the relationship between the use of statins and urinary albumin excretion (UAE) and glomerular filtration rate (GFR) in two settings: a randomized controlled trial (RCT) and an observational cohort study, in which patients were included to study the impact of an elevated UAE on renal and cardiovascular prognosis. METHODS: We used data from the Prevention of REnal and Vascular ENd-stage Disease Intervention trial (PREVEND-IT) and the PREVEND cohort study. The PREVEND-IT subjects (788 with a UAE 15-300 mg/day) received pravastatin 40 mg/day vs placebo and/or fosinopril 20 mg/day vs placebo in a 2x2 factorial-RCT design. Of the 3440 cohort subjects, 469 used statins during the 4 year follow-up period. Multivariate-regression adjusted for confounding factors and the propensity score was used to estimate the relation between statin use and UAE and GFR. RESULTS: In the RCT, pravastatin did not change UAE or GFR, neither in fosinopril yes/no subgroups. In the observational cohort, statin use was associated with a rise in UAE (+12.1%), compared with statin non-use (+3.6%, P<0.001). This rise was most pronounced in those on statins prior to the first screening [+24.8% (95% CI: 11.9-39.2)], those using statins>3 years [+18.5% (7.3 30.8)] and those with >1 or >2 defined daily doses (+15.7 and +17.3%, respectively). These differences remained significant after adjustment for relevant variables and propensity score. The rise in UAE could not be attributed to a higher dose or a specific statin. GFR fell in 4 years in both statin users and non-users (4.6+/-13.5 and 2.4+/-11.2, respectively). The fall in GFR between groups was not different after adjustment (P=0.11). CONCLUSIONS: We conclude from the RCT data that statins do not lower UAE in subjects selected because of an elevated UAE instead of hyperlipidaemia. In the observational cohort study, the use of statins similarly was not associated with a fall in UAE; UAE instead increased. Statin treatment was not associated with a significant change in GFR in these subjects with only modestly impaired GFR. PMID- 16720595 TI - Differences in decline in GFR with age between males and females. Reference data on clearances of inulin and PAH in potential kidney donors. AB - BACKGROUND: To ensure that potential kidney donors have no renal impairment, it is extremely important to have accurate methods for evaluating the glomerular filtration rate (GFR). The golden standard, clearance of inulin, has been used in the present study. The aim was to evaluate the effects of age and sex on renal function and present reference data. METHODS: A total of 122 potential kidney donors, 62 females, aged 21-67 years, were investigated with the GFR and effective renal plasma flow (ERPF) determined by clearances of inulin and para amino hippurate. RESULTS: The mean +/- SD GFR and ERPF were 105 +/- 13 and 545 +/ 108 ml/min/1.73 m(2), respectively, and we found no difference between the males and females. When relating GFR and ERPF to age, however, a significant decline was found in GFR and ERPF in males, but not in females in the age range of 20-50 years. GFR fell by a mean of 8.7 ml/min/1.73 m(2) and ERPF by 90 ml/min/1.73 m(2) per decade in male donors. CONCLUSION: With adequate methods for determining GFR and ERPF, a clear difference in the effect of age was seen between the sexes. Males showed a significant decrease between 20 and 50 years of age, which was not seen in females. Females seem to be protected in the pre-menopausal period probably by oestrogens. These results confirm clinically those found in rats. PMID- 16720594 TI - The role of oral dryness in interdialytic weight gain by diabetic and non diabetic haemodialysis patients. AB - BACKGROUND: Factors influencing the percentage of daily interdialytic weight gain (IDWG%) and their interactions in haemodialysis (HD) patients have not been well defined, especially in diabetic patients. We analysed contributing factors for the increase of IDWG%, particularly xerostomia (oral dryness), among diabetic and non-diabetic HD patients. METHODS: We collected 3 month prospective data in 184 stable HD patients (116 non-diabetic and 68 diabetic), including assessments of xerostomia by 100 mm visual analog scales (VASs), and the unstimulated whole salivary (UWS) flow rate was measured in 91 patients by a spitting method. RESULTS: Diabetic patients have higher IDWG% (P = 0.042) and VAS oral dryness score (P = 0.021), whereas, have lower UWS (P = 0.032). In non-diabetic patients, the VAS oral dryness score, age, Kt/V and blood urea nitrogen (BUN) level correlated independently with IDWG%. In diabetic patients, the haemoglobin A(1C) (HbA(IC)) correlated significantly with IDWG% after controlling for age, Kt/V and BUN level; however, when VAS oral dryness score was introduced into the regression model, the effect of HbA(IC) became marginally significant (P = 0.073) while the VAS oral dryness score became significantly correlated with IDWG%. The increases in IDWG% per unit change in VAS oral dryness score did not show significant difference between the non-diabetic and total diabetic patients; however, it was larger in patients with HbA(IC) >or=9%. CONCLUSIONS: Xerostomia plays a significant role in increasing IDWG% among diabetic and non-diabetic HD patients. In diabetic patients, the increased IDWG% associated with the increasing HbA(1C) level is largely dependent on the severity of xerostomia, and we speculate that insulin deficiency may operate synergistically with xerostomia in increasing IDWG% in patients with HbA(1C) >or=9%. PMID- 16720596 TI - PPAR-alpha and -gamma agonists attenuate diabetic kidney disease in the apolipoprotein E knockout mouse. AB - BACKGROUND: Peroxisome proliferator-activated receptor (PPAR)-alpha and PPAR gamma agonists are widely used in diabetes. In addition to their effects on lipid and glucose homeostasis, these agents have been postulated to have independent renoprotective actions. In the current study, we assess the efficacy of the PPAR alpha agonist, gemfibrozil, the PPAR-gamma agonist rosiglitazone and the non thiazolidinedione PPAR-alpha/gamma coagonist, compound 3q, on kidney structure and function in streptozotocin-treated apolipoprotein E knockout mice. METHODS: Control and streptozotocin-diabetic mice were randomized to receive rosiglitazone (20 mg/kg/day), gemfibrozil (100 mg/kg/day), or compound 3q (3 mg/kg/day) by gavage, or no treatment for a period of 20 weeks. Renal fibrosis was assessed by standard histology and collagen IV immunohistochemistry. Kidney function was assessed by urinary albumin excretion and creatinine clearance. RESULTS: Diabetes in this model was associated with an increase in glomerulosclerosis, tubulointerstitial fibrosis and increased collagen IV deposition in the glomeruli and tubules. All three agents significantly attenuated glomerulosclerosis, tubulointerstitial expansion and collagen IV deposition. The increase in albuminuria and the decline in kidney function associated with diabetes in this model were also attenuated by each of these agents, with no superiority observed among various treatment groups. These renoprotective effects were observed in the absence of changes in glucose, insulin or lipid levels or a reduction in blood pressure. CONCLUSIONS: Combined with their independent anti-atherosclerotic actions, and their important effects on dyslipidaemia and insulin resistance, PPAR agonists may be useful for the prevention of diabetic complications, including kidney disease, even in type 1 diabetes. PMID- 16720597 TI - Haploinsufficiency of Pkd2 is associated with increased tubular cell proliferation and interstitial fibrosis in two murine Pkd2 models. AB - BACKGROUND: Autosomal dominant polycystic kidney disease (ADPKD) is the most common inherited human kidney disease and is caused by germline mutations in PKD1 (85%) or PKD2 (15%). It has been estimated that around 1% of tubular cells give rise to cysts, and cell hyperproliferation has been noted to be a cardinal feature of cystic epithelium. Nevertheless, it is uncertain whether the increase in proliferative index observed is an early or late feature of the cystic ADPKD kidney. METHODS: Two Pkd2 mouse mutants (WS25 and WS183) have been recently generated as orthologous models of PKD2. To determine the effect of Pkd2 dosage on cell proliferation, cyst formation and renal fibrosis, we studied renal tissue from Pkd2(WS25/WS25) and Pkd2(+/-) mice by histological analysis. We also examined the proliferative index in archival nephrectomy tissue obtained from patients with ADPKD and normal controls. RESULTS: The proliferative index of non cystic tubules in Pkd2 mutant mice as assessed by proliferating cell nuclear antigen and Ki67-positive nuclei was between 1-2%, values 5-10 times higher than control tissue. Similarly, the proliferative index of non-cystic tubules in human ADPKD kidneys was 40 times higher than corresponding controls. In Pkd2 mutant mice, significant correlations were found between the fibrosis score and the mean cyst area as well as with the proliferative index. Of significance, proliferating tubular cells were uniformly positive for polycystin-2 expression in Pkd2(+/-) kidney. CONCLUSION: These results suggest that an increase in cell proliferation is an early event preceding cyst formation and can result from haploinsufficiency at Pkd2. The possible pathogenic link between tubular cell proliferation, interstitial fibrosis and cyst formation is discussed. PMID- 16720598 TI - Reduced graft function (with or without dialysis) vs immediate graft function--a comparison of long-term renal allograft survival. AB - BACKGROUND: Delayed graft function (DGF) is a common complication in cadaveric kidney transplants affecting graft outcome. However, the incidence of DGF differs widely between centres as its definition is very variable. The purpose of this study was to define a parameter for DGF and immediate graft function (IGF) and to compare the graft outcome between these groups at our centre. METHODS: The renal allograft function of 972 first cadaveric transplants performed between 1990 and 2001 in the Republic of Ireland was examined. The DGF and IGF were defined by a creatinine reduction ratio (CRR) between time 0 of transplantation and day 7 post transplantation of <70 and >70%, respectively. Recipients with reduced graft function (DGF) not requiring dialysis were defined as slow graft function (SGF) patients. The serum creatinine at 3 months, 6 months, 1, 2 and 5 years after transplantation was compared between these groups of recipients. The graft survival rates at 1, 3 and 5 years and the graft half-life for DGF, SGF and IGF recipients were also assessed. RESULTS: Of the 972 renal transplant recipients, DGF was seen in 102 (10.5%) patients, SGF in 202 (20.8%) recipients and IGF in 668 (68.7%) patients. Serum creatinine levels were significantly different between the three groups at 3 and 6 months, 1, 2 and 5 years. Graft survival at 5 years for the DGF patients was 48.5%, 60.5% for SGF recipients and 75% for IGF patients with graft half-life of 4.9, 8.7 and 10.5 years, respectively. CONCLUSION: This study has shown that the CRR at day 7 correlates with renal function up to 5 years post-transplantation and with long-term graft survival. We have also demonstrated that amongst patients with reduced graft function after transplantation, two groups with significantly different outcomes exist. PMID- 16720599 TI - Granulomatous interstitial nephritis treated with a tumour necrosis factor-alpha inhibitor. PMID- 16720600 TI - Style-by-style analysis of two sporadic self-compatible Solanum chacoense lines supports a primary role for S-RNases in determining pollen rejection thresholds. AB - A method for the quantification of S-RNase levels in single styles of self incompatible Solanum chacoense was developed and applied toward an experimental determination of the S-RNase threshold required for pollen rejection. It was found that, when single style values are averaged, accumulated levels of the S(11)- and S(12)-RNases can differ up to 10-fold within a genotype, while accumulated levels of the S(12)-RNase can differ by over 3-fold when different genotypes are compared. Surprisingly, the amount of S(12)-RNase accumulated in different styles of the same plant can differ by over 20-fold. A low level of 160 ng S-RNase in individual styles of fully incompatible plants, and a high value of 68 ng in a sporadic self-compatible (SSC) line during a bout of complete compatibility was measured, suggesting that these values bracket the threshold level of S-RNase needed for pollen rejection. Remarkably, correlations of S-RNase values to average fruit sets in different plant lines displaying sporadic self compatibility (SSC) to different extents as well as to fruit set in immature flowers, are all consistent with a threshold value of 80 ng S(12)-RNase. Taken together, these results suggest that S-RNase levels alone are the principal determinant of the incompatibility phenotype. Interestingly, while the S-RNase threshold required for rejection of S(12)-pollen from a given genetic background is the same in styles of different genetic backgrounds, it is different when pollen donors of different genetic backgrounds are used. These results reveal a previously unsuspected level of complexity in the incompatibility reaction. PMID- 16720601 TI - Modulation of plant morphology, root architecture, and cell structure by low vitamin C in Arabidopsis thaliana. AB - Ascorbic acid (AA) fulfils many essential functions in plants. It is a key antioxidant and an important reducing substrate for a number of enzymes. The effects of low AA on plant architecture and leaf ultrastructure were studied in Arabidopsis thaliana mutants, which have constitutively moderately low (vtc1) or very low (vtc2) leaf AA contents compared with the wild type. Shoot development was comparable in all accessions over the first 14 d of growth. The production of primary roots was slightly different in vtc1, vtc2, and wild-type plants. However, the most notable difference was that a high proportion of the primary roots of the vtc2 plants grown on soil had lost the wild-type responses to gravity. The vtc mutants showed the antagonistic interaction between nitrate and sugar in the regulation of lateral root (LR) development that was observed in the wild type. However, the vtc2 mutants produced greater numbers of longer LRs than wild-type or vtc1 plants at all levels of nitrate. At later stages of development, the vtc rosettes were smaller than those of the wild type and the leaves showed intracellular structural changes that are consistent with programmed cell death (PCD). PCD symptoms such as nuclear chromatin condensation, the presence of multivesicular bodies, and extensive degradation and disorganization of the grana stacks were observed in 8-week-old vtc2 leaves and in 10-week-old vtc1 leaves. The data presented here illustrate the importance of tissue AA contents in regulating whole plant morphology, cell structure, and development. PMID- 16720602 TI - Genome-wide analysis of plant glutaredoxin systems. AB - The recent release of the first tree genome (Populus trichocarpa) has allowed a comparison to be made of the multigenic glutaredoxin (Grx) and glutathione reductase (GR) families of this tree with those of other sequenced organisms and especially of the two other fully sequenced plant species, Arabidopsis thaliana and Oryza sativa. Grxs are small proteins involved in disulphide bridge or protein-glutathione adduct reduction, and they are maintained in a reduced form using glutathione and an NADPH-dependent GR. While the P. trichocarpa and O. sativa genomes are nearly five times larger than that of A. thaliana, they contain approximately 45 000 and 37 500 genes compared with the 25 500 genes of A. thaliana. On the one hand, the GR gene composition varies little between species and the gene structures are relatively conserved. On the other hand, the Grx gene family can be divided into three subgroups and the gene content is larger in P. trichocarpa (36 genes) compared with A. thaliana and O. sativa (31 and 27 genes, respectively). This could be partly explained by the occurrence of more duplication events, and this is especially true for one of the three identified Grx subgroups (subgroup III). The expression of most of these genes was confirmed by analysing expressed sequence tags present in various databases. In addition, the expression of Grx of subgroups I and II was examined by RT-PCR in various poplar organs. A complete classification based essentially on gene structure and sequence identity is proposed. PMID- 16720603 TI - Production of reactive oxygen species in Arabidopsis thaliana cell suspension cultures in response to an elicitor from Fusarium oxysporum: implications for basal resistance. AB - The present understanding of ROS generation in the defence response of Arabidopsis thaliana is reviewed. Evidence suggests that the apoplastic oxidative burst generated during basal resistance is peroxidase-dependent. The ROS generated during this basal resistance may serve to activate NADPH oxidase during the R-gene-mediated hypersensitive response. The processes involved in the production of reactive oxygen species in A. thaliana cell suspension cultures in response to an elicitor from Fusarium oxysporum are investigated in the present work. This system appears analogous to the production of ROS during the basal resistance response in French bean, which is peroxidase-dependent. A panel of modulators effective in other pathogen elicitor and plant cell systems has been used to investigate the Arabidopsis signalling pathways and the plant cell responses involved. Thus as in other systems, an early calcium influx into the cytosolic compartment, a rapid efflux of K(+) and Cl(-), and extracellular alkalinization of elicited cell cultures has been found. However the alkalinization is not sufficient to stimulate the apoplastic oxidative burst by itself, unlike in French bean, although vectorial ion fluxes are needed. A secretory component which is sensitive to monensin and N-ethylmaleimide and insensitive to brefeldin A may also be necessary for the release and provision of substrates for peroxidase-dependent generation of H(2)O(2). PMID- 16720604 TI - The role of reactive oxygen species in cell growth: lessons from root hairs. AB - Reactive oxygen species (ROS) play a diversity of roles in plants. In recent years, a role for NADPH oxidase-derived ROS during cell growth and development has been discovered in a number of plant model systems. These studies indicate that ROS are required for cell expansion during the morphogenesis of organs such as roots and leaves. Furthermore, there is evidence that ROS are required for root hair growth where they control the activity of calcium channels required for polar growth. The role of ROS in the control of root hair growth is reviewed here and results are highlighted that may provide insight into the mechanism of plant cell growth in general. PMID- 16720605 TI - A genetic approach towards elucidating the biological activity of different reactive oxygen species in Arabidopsis thaliana. AB - Plants are often exposed to external conditions that adversely affect their growth, development or productivity. Such unfavourable environmental stress factors may result in rapid and transient increases of intracellular concentrations of reactive oxygen species (ROS) that are chemically distinct and impact plants either by being cytotoxic or by acting as a signal. Because different ROS are generated simultaneously in different cellular and extracellular compartments, it is almost impossible to link a particular ROS to a specific stress response and to determine its mode of action. The conditional flu mutant of Arabidopsis has been used to determine the biological role of singlet oxygen. Immediately after a dark/light shift of the flu mutant, singlet oxygen is generated within the plastids activating several stress responses that include growth inhibition of mature plants and seedling lethality. These stress responses do not result from physicochemical damage caused by singlet oxygen, but are attributable to the activation of a genetically determined stress response programme triggered by the Executer1 protein. Singlet oxygen-mediated stress responses at the transcriptional level necessitate a retrograde transduction of signals from the chloroplast to the nucleus that activate distinct sets of genes different from those that are induced by superoxide/hydrogen peroxide. Hence, the biological activities of these two types of ROS are distinct from each other. Whether they act independently or interact is not known yet and is the topic of our current research. PMID- 16720606 TI - Effect of eyespot caused by Oculimacula yallundae and O. acuformis, assessed visually and by competitive PCR, on stem strength associated with lodging resistance and yield of winter wheat. AB - Winter wheat, (cv. Consort) was inoculated with three isolates of either Oculimacula yallundae or O. acuformis to determine the effect of eyespot caused by each species on yield and lodging resistance of winter wheat. Plants were visually assessed for disease incidence and severity, and pathogen DNA was quantified at GS 33 and GS 60. At early milk development of the crop (GS 72), 900 main shoots were also visually assessed for the disease and subjected to mechanical tests for stem strength. Pathogen DNA was extracted from each shoot and quantified using competitive polymerase chain reaction (PCR). Although slight and moderate eyespot lesions caused by either species had no effect on ear weight, severe lesions caused by O. acuformis and O. yallundae reduced ear weight by 3% and 7%, respectively. Stem lodging failed to occur at the site; however, yield losses of 11% for O. acuformis and 6% for O. yallundae were observed. Visual assessment failed to reveal differences between species in their effect on plant characteristics, stem bending strength, or stem safety factor. PCR data, however, showed that the two species had similar effects determined by different DNA concentrations. Both species reduced lodging resistance (stem safety factor) compared with the control. In contrast to healthy plants, where reductions were related predominantly to the height and weight distribution of the plants, the observed reductions of stem lodging resistance in infected plants with Oculimacula spp. were associated primarily with reductions in stem bending strength. PMID- 16720607 TI - An evolutionary perspective on the regulation of carpel development. AB - The carpel, or female reproductive organ enclosing the ovules, is one of the major evolutionary innovations of the flowering plants. The control of carpel development has been intensively studied in the model eudicot species Arabidopsis thaliana. This review traces the evolutionary history of genes involved in carpel development by surveying orthologous genes in taxa whose lineages separated from that of A. thaliana at different levels of the phylogenetic tree of the seed plants. Some aspects of the control of female reproductive development are conserved between the flowering plants and their sister group, the gymnosperms, indicating the presence of these in the common ancestor of the extant seeds plants, some 300 million years ago. Gene duplications that took place in the pre angiosperm lineage, before the evolution of the first flowering plants, provided novel gene clades of potential importance for the origin of the carpel. Subsequent to the appearance of the first flowering plants, further gene duplications have led to sub-functionalization events, in which pre-existing reproductive functions were shared between paralogous gene clades. In some cases, fluidity in gene function is evident, leading to similar functions in carpel development being controlled by non-orthologous genes in different taxa. In other cases, gene duplication events have created sequences that evolved novel functions by the process of neo-functionalization, thereby generating biodiversity in carpel and fruit structures. PMID- 16720608 TI - Isolation of a novel ABC-transporter gene from soybean induced by salicylic acid. AB - This paper reports on the identification and characterization of a new ATP binding cassette (ABC) transporter which was identified as a salicylic acid induced gene from soybean (Glycine max cv. Williams 82) in a subtractive suppression hybridization approach. A fragment of an ABC-transporter gene was used to isolate a full-length cDNA clone for this gene with a length of 4750 bp. The encoded protein has a length of 1447 amino acids and is composed of two similar repeat units typical of full-size ABC transporters. The sequence displays a close relationship to plant pleiotropic drug resistance (PDR)-type transporters and, on a homology basis, clusters together with the Arabidopsis thaliana PDR12 gene, suggesting GmPDR12 as a name for the gene isolated from soybean. GmPDR12 is rapidly responsive to salicylic acid and methyl jasmonate. The mRNA starts to accumulate 30 min after the addition of the signalling compounds. Salicylic acid is required for the execution of the hypersensitive reaction in soybean cell suspension cultures inoculated with Pseudomonas syringae pv. glycinea. It has been demonstrated previously that salicylic acid can be substituted by a variety of functional analogues of salicylic acid. All of these compounds lead to a strong and rapid transcriptional activation of GmPDR12, suggesting a shared signalling pathway. PMID- 16720609 TI - Cellulose orientation determines mechanical anisotropy in onion epidermis cell walls. AB - The role of cellulose microfibril orientation in determining cell wall mechanical anisotropy and in the control of the wall plastic versus elastic properties was studied in the adaxial epidermis of onion bulb scales using the constant-load (creep) test. The mean or net cellulose orientation in the outer periclinal wall of the epidermis was parallel to the long axis of the cells. In vitro cell wall extensibility was 30-90% higher in the direction perpendicular to the net microfibril orientation than parallel to it. This was the case for the size of the initial deformation occurring just after the load application and for the rate of time-dependent creep. Loading/unloading experiments confirmed the presence of a real irreversible component in cell wall extension. The plastic component of the time-dependent deformation was higher perpendicular to the net cellulose orientation than parallel to it. An acid buffer (pH 4.5) increased the creep rate by 25-30% but this response was not related to cellulose orientation. The present data provide direct evidence that the net orientation of cellulose microfibrils confers mechanical anisotropy to the walls of seed plants, a characteristic that may be relevant to understanding anisotropic cell growth. PMID- 16720610 TI - Effects of simultaneous expression of heterologous genes involved in phytochelatin biosynthesis on thiol content and cadmium accumulation in tobacco plants. AB - Transgenic tobacco (Nicotiana tabacum cv. LA Burley 21) lines expressing three genes encoding enzymes thought to be critical for the efficient production of phytochelatins, (i) serine acetyltransferase (EC 2.3.1.30) involved in the production of O-acetylserine, the cysteine precursor, (ii) gamma-glutamylcysteine synthetase (EC 6.3.2.2) involved in the production of gamma-glutamylcysteine, the precursor of glutathione, and (iii) phytochelatin synthase (EC 2.3.2.15), were obtained and analysed for non-protein thiol content and cadmium accumulation. After a 3 week exposure to 15 microM CdCl2, plants expressing transgenes (either separately or in combination) had increased cadmium concentration in roots but not in shoots compared with the wild type. Nearly all transgenic lines analysed had more non-protein thiols than the wild type. The greatest effects (about 8 fold elevation of thiols) were found in one of the lines simultaneously expressing the three transgenes. Despite the fact that a multi-transgene strategy described in this work resulted in a strong increase in the levels of several classes of non-protein thiols in transgenic plants, other factors appeared to restrict cadmium accumulation in shoots. PMID- 16720611 TI - Cloning and characterization of three thioredoxin h isoforms from wheat showing differential expression in seeds. AB - Plants contain several genes encoding thioredoxin h. In cereals, type-h thioredoxins are abundant in developing and germinating grains, but the mechanism regulating the expression of these genes and their specific function is poorly known. The cloning of three full-length cDNAs encoding thioredoxin h, stated Trxh1, Trxh2 and Trxh3, from wheat (Triticum aestivum cv. Soissons) seeds is described here. TRXh2 and TRXh3 deduced proteins show high identity between them and with other thioredoxins h previously described from wheat, and contain exclusively the two Cys residues forming part of the active site. By contrast, TRXh1 shows a lower level of identity and contains an additional Cys residue. The three wheat thioredoxins were expressed in E. coli and their activity was demonstrated using both the DTT-dependent insulin assay and a coupled assay with recombinant NTR from wheat. Site-directed mutagenesis showed that the additional Cys residue of TRXh1 has a low effect on its activity but is essential for dimerization. Specific expression of the three thioredoxin genes was analysed by real-time RT-PCR in developing and germinating seeds and seedlings under stressed and unstressed conditions. An increase of Trxh1, Trxh2, and Trxh3 transcripts was detected at the beginning of the desiccation phase during seed development. Early after imbibition, Trxh1, but not Trxh2 or Trxh3, transcripts showed a transient increase. Treatment of wheat seedlings with salt or hydrogen peroxide caused a differential pattern of expression of the three Trxh genes between and within tissues, hence suggesting specific functions for these thioredoxins during germination and early seedling growth. PMID- 16720612 TI - Localization of arabinogalactan proteins in egg cells, zygotes, and two-celled proembryos and effects of beta-D-glucosyl Yariv reagent on egg cell fertilization and zygote division in Nicotiana tabacum L. AB - Arabinogalactan proteins (AGPs) have been implicated in a variety of plant development processes including sexual plant reproduction. As a crucial developmental event, plant sexual reproduction generally occurs inside an ovule embedded in an ovary. The inaccessibility of the egg cells, zygotes, and embryos has hindered our understanding of the importance of AGPs in the early events involving fertilization, zygotic division, and early embryogenesis. In this study, the well-established in vitro zygote and ovary culture systems, together with immunofluorescence and immunogold labelling techniques, were employed to investigate the role of AGPs in the early events of sexual reproduction in Nicotiana tabacum. Dramatic changes in AGP content during ovule development were evidenced by western blotting. Subcellular localization revealed that AGPs are localized in the plasma membrane, cell wall, and cytoplasm of pre- and post fertilized egg cells, and cytoplasm and vacuoles of two-celled proembryos. Abundant AGPs were detected in unfertilized egg cells; however, the level of AGPs substantially decreased in fertilized egg cells. Polar distribution of AGPs in elongated zygotes was observed. The early two-celled proembryos just from zygote division displayed accumulation of AGPs at a low level, while in the elongated two-celled proembryos at the late stage, the AGP content clearly increased. Provision of betaGlcY, a synthetic phenylglycoside that specifically binds AGPs, to the in vitro cultures of isolated zygote and fertilized ovaries increased abnormal symmetrical division of zygotes. In the culture of pollinated but unfertilized ovaries, addition of betaGlcY resulted in arrest of fertilization of the egg cells, but had no effect on fertilization of the central cells. The possible roles of AGPs in fertilization, zygotic division, and proembryo development are discussed. PMID- 16720613 TI - Transcriptomic analysis indicates putative metabolic changes caused by manipulation of phosphorus availability in rice leaves. AB - Plants have developed several strategies for coping with phosphorus (P) deficiency. However, the details of the regulation of gene expression of adaptations to low P are still unclear. Using a cDNA microarray, transcriptomic analyses were carried out of the rice genes regulated by P deficiency and P re supply to P-deficient plants. The OsPI1 gene, which was isolated as the most significant up-regulated gene under -P conditions, was also the most significant down-regulated gene following P re-supply. Many starch metabolism-related genes, as well as several genes for P(i)-liberating enzymes, were up-regulated by -P treatment, suggesting a homeostatic contribution to the P(i) concentration in leaf tissues. mRNAs for glucanases were also induced by P re-supply: these are suspected to play a role in loosening the cell wall compounds. Most of the genes up-regulated by -P treatment were down-regulated by P re-supply, suggesting that their responses were specific to -P conditions. Conversely, the number of genes up-regulated by P re-supply was also larger following P re-supply than in the -P condition. It is proposed that the genes up-regulated by P re-supply play an important role in P acquisition by P-deficient plants. PMID- 16720614 TI - The contribution of extensin network formation to rapid, hydrogen peroxide mediated increases in grapevine callus wall resistance to fungal lytic enzymes. AB - Grapevine (Vitis vinifera cv. Touriga) callus cell walls contain a high level of the monomeric extensin, GvP1. Hydrogen peroxide stimulus of these cultures causes the rapid loss of monomeric GvP1, concomitant with marked increases in insoluble GvP1 amino acids and wall resistance to digestion by fungal lytic enzymes. JIM11 immunolocalization studies indicated that monomeric and network GvP1 were evenly distributed in the callus cell wall. These primary cell walls were used to investigate the specific contribution of extensin and other ionically bound cell wall proteins to hydrogen peroxide-mediated increases in resistance to fungal lytic enzymes. This was performed by removing ionically-bound proteins and assaying for hydrogen peroxide-enhanced resistance after the addition of selected protein fractions. The results indicate that hydrogen peroxide-induced increases in resistance to digestion by fungal lytic enzymes require a co-operative action between network extensin formation and the electrostatic interaction of additional wall proteins with the extracellular matrix. PMID- 16720615 TI - The expression patterns of SAG12/Cab genes reveal the spatial and temporal progression of leaf senescence in Brassica napus L. with sensitivity to the environment. AB - Despite a high nitrate uptake capacity, the nitrogen use efficiency (NUE) of oilseed rape is weak due to a relatively low N remobilization from vegetative (mostly leaves) to growing parts of the plant. Thus, this crop requires a high rate of N fertilization and leaves fall with a high N content. In order to reduce the rate of N fertilization and to improve the environmental impact of oilseed rape, new genotypes could be selected on their capacity to mobilize the foliar N. Various indicators of leaf senescence in oilseed rape were analysed during plant growth, as well as during senescence induced by N deprivation. Metabolic changes in leaves of increasing age were followed in N-supplied and N-deprived rosettes by measuring chlorophyll, total N, and soluble protein contents. Similarly, the expression of genes known to be up-regulated (SAG12) or down-regulated (Cab) during leaf senescence was monitored. The amount of soluble proteins per leaf was a better indicator of leaf senescence than chlorophyll or total N content, but was not evaluated as an accurate indicator under conditions of N deprivation. On the other hand, up-regulation of SAG12 concomitantly with down-regulation of Cab in the leaf revealed the spatial and temporal progression of leaf senescence in oilseed rape. This study shows, for the first time at the whole plant level, that the SAG12/Cab gene expressions match the sink/source transition for N during both developmental and nutrient stress-induced leaf senescence. PMID- 16720616 TI - Emerging possibilities in the development and function of regulatory T cells. AB - CD25+CD4+ Regulatory T cells (Treg) represent a unique population of lymphocytes capable of powerfully suppressing immune responses. A large body of experimental data have now confirmed the essential role played by these cells in a host of clinically relevant areas such as self-tolerance, transplantation, allergy and tumor/microbial immunity. Despite this mass of knowledge, significant gaps in our understanding of fundamental Treg biology remain, particularly regarding their development and mechanisms of suppression. In this review we attempt to highlight the current controversies and directions in which this exciting field is moving. PMID- 16720617 TI - Missense mutations in SH2D1A identified in patients with X-linked lymphoproliferative disease differentially affect the expression and function of SAP. AB - X-linked lymphoproliferative disease (XLP) is an immunodeficiency resulting from mutations in SH2D1A, which encodes signalling lymphocytic activation molecule (SLAM)-associated protein (SAP). In addition to SLAM, SAP associates with several other cell-surface receptors including 2B4 (CD244), Ly9 (CD229), CD84 and NTB-A. SAP contains a single src-homology-2 domain and acts as an intracellular adaptor protein by recruiting the protein tyrosine kinase FynT to the cytoplasmic domains of some of these receptors, which results in the initiation of specific downstream signal transduction pathways. XLP is likely to result from perturbed signalling through one or more of these SAP-associating receptors. In this study, we identified missense (Y54C, I84T and F87S) and insertion (fs82 --> X103) mutations in four different kindreds affected by XLP. Each mutation dramatically reduced the half-life of SAP, thus diminishing its expression in primary lymphocytes as well as in transfected cell lines. Interestingly, although the Y54C and F87S mutations compromised the ability of SAP to associate with different receptors, the I84T mutation had no effect on the ability of SAP to bind SLAM, CD84 or 2B4. However, signalling downstream of SLAM was reduced in the presence of SAP bearing the I84T mutation. These findings indicate that, irrespective of the type of mutation, signalling through SAP-associating receptors in XLP can be impaired by reducing the expression of SAP, the ability of SAP to bind surface receptors and/or its ability to activate signal transduction downstream of the SLAM-SAP complex. PMID- 16720618 TI - CD38 induces apoptosis of a murine pro-B leukemic cell line by a tyrosine kinase dependent but ADP-ribosyl cyclase- and NAD glycohydrolase-independent mechanism. AB - Cross-linking of CD38 on hematopoietic cells induces activation, proliferation and differentiation of mature T and B cells and mediates apoptosis of myeloid and lymphoid progenitor cells. In addition to acting as a signaling receptor, CD38 is also an enzyme capable of producing several calcium-mobilizing metabolites, including cyclic adenosine diphosphate ribose (cADPR). It has been previously postulated that the calcium-mobilizing metabolites produced by CD38 may regulate its receptor-based activities. To test this hypothesis, we examined whether the enzyme activity of CD38 controls the apoptosis of an anti-CD38-stimulated leukemic B cell. We show that anti-CD38-induced apoptosis of Ba/F3 cells, a murine pro-B cell line, is not affected by blocking the calcium-mobilizing activity of cADPR or by inhibiting intracellular or extracellular calcium mobilization. In addition, we demonstrate that blocking CD38 enzyme activity with 2'-deoxy-2'-fluoro-nicotinamide arabinoside adenine dinucleotide has no effect on apoptosis and that Ba/F3 cells expressing catalytically inactive mutant forms of CD38 still undergo apoptosis upon CD38 cross-linking. Instead, we find that anti CD38-induced apoptosis is dependent on tyrosine kinase and caspase activation, and that this process appears to be potentiated by the presence of membrane microdomains. Thus, the receptor-mediated functions of CD38 can be separated from its enzyme activity in a murine leukemic cell line, suggesting that CD38 plays multiple, but independent, biologic roles. PMID- 16720619 TI - Randomised controlled trial of four commercial weight loss programmes in the UK: initial findings from the BBC "diet trials". AB - OBJECTIVE: To compare the effectiveness of four commercial weight loss diets available to adults in the United Kingdom. DESIGN: Six month multicentre randomised unblinded controlled trial. SETTING: Community based sample of otherwise healthy overweight and obese adults. INTERVENTIONS: Dr Atkins' new diet revolution, Slim-Fast plan, Weight Watchers pure points programme, and Rosemary Conley's eat yourself slim diet and fitness plan. MAIN OUTCOME MEASURES: Weight and body fat changes over six months. RESULTS: All diets resulted in significant loss of body fat and weight over six months. Groups did not differ significantly but loss of body fat and weight was greater in all groups compared with the control group. In an intention to treat analysis, average weight loss was 5.9 kg and average fat loss was 4.4 kg over six months. The Atkins diet resulted in significantly higher weight loss during the first four weeks, but by the end was no more or less effective than the other diets. CONCLUSIONS: Clinically useful weight loss and fat loss can be achieved in adults who are motivated to follow commercial diets for a substantial period. Given the limited resources for weight management in the NHS, healthcare practitioners should discuss with their patients programmes known to be effective. TRIAL REGISTRATION: Clinical trials NCT00327821. PMID- 16720620 TI - Biologically active molecules that reduce polyglutamine aggregation and toxicity. AB - Polyglutamine expansion in certain proteins causes neurodegeneration in inherited disorders such as Huntington disease and X-linked spinobulbar muscular atrophy. Polyglutamine tracts promote protein aggregation in vitro and in vivo with a strict length-dependence that strongly implicates alternative protein folding and/or aggregation as a proximal cause of cellular toxicity and neurodegeneration. We used an intracellular polyglutamine protein aggregation assay based on fluorescence resonance energy transfer (FRET) to identify inhibitors of androgen receptor (AR) aggregation in three libraries of biologically active small molecules: the Annotated Compound Library, the NINDS Custom Collection and a kinase inhibitor collection. In the primary screen 10 compounds reduced AR aggregation. While 10/10 also reduced huntingtin (Htt) exon 1 aggregation, only 2/10 reduced aggregation of pure polyglutamine peptides. In a PC-12 model 9/10 compounds reduced aggregation. Five out of nine compounds tested in an Htt exon 1 assay of neurodegeneration in Drosophila partially rescued the phenotype. Three of the five compounds effective in flies are FDA-approved drugs. These compounds provide new leads for therapeutic development for the polyglutamine diseases based on their efficacy in mammalian cells and a Drosophila model. The high predictive value of the primary screen suggests that the FRET-based screening assay may be useful for further primary and secondary screens for genes or small molecules that inhibit polyglutamine protein aggregation. PMID- 16720621 TI - Hyperemesis gravidarum: is an ultrasound scan necessary? AB - BACKGROUND: Traditionally, in cases of hyperemesis gravidarum (HG), an ultrasound evaluation is recommended to confirm viability and to exclude multiple pregnancies and gestational trophoblastic disease (GTD). Our aim was to perform a case-control study to evaluate the incidence of these findings. METHODS: Each case of HG was matched for gestational age with the next ultrasound examination performed in an asymptomatic pregnancy. The findings were compared between the two groups. RESULTS: Two hundred and eighty-six cases of HG were matched with 286 asymptomatic women. The total number of viable pregnancies was higher in the HG group (280/286, 97.9%) than that in the control group (265/286, 92.6%; P = 0.006). The incidence of twins was 3.1% in each group (P > 0.999). The incidence of early pregnancy failure was 0.7% in women with HG compared to 7.0% in asymptomatic women (odds ratio 0.09, 95% CI 0.01-0.04, P < 0.0001). The one case of GTD was in the HG group; however, this case also presented with vaginal bleeding. CONCLUSIONS: Pregnancies complicated by HG had a similar risk of twin pregnancy, and a lower risk of early pregnancy failure compared to controls. In the absence of vaginal bleeding, there was no increase in GTD in women with HG. We conclude that an ultrasound scan is not clinically necessary in women presenting with HG, other than for maternal reassurance. PMID- 16720622 TI - Anti-Mullerian hormone inhibits initiation of growth of human primordial ovarian follicles in vitro. AB - BACKGROUND: Anti-Mullerian hormone (AMH) inhibits the initiation of the development and early growth of mouse ovarian follicles. Furthermore, the ovarian follicle pool diminishes prematurely in AMH-knockout mice. In this study, we examined whether AMH plays a similar role in humans, controlling ovarian follicle growth. METHODS: Human ovarian cortical tissue biopsy specimens were cut into small pieces and cultured for 7 days in medium containing rat recombinant AMH at 0, 10, 30 or 100 ng/ml. The developmental stages and viability of the follicles were evaluated from histological sections. RESULTS: Similar to previous studies, significant initiation of follicle growth was observed in almost all culture media, as demonstrated by a significantly smaller proportion of primordial follicles (14-26%) compared with non-cultured control tissue (56%). The exception was tissue in medium supplemented with AMH at 100 ng/ml. Here, the proportion of primordial follicles was not significantly different from that in non-cultured tissue; furthermore, it was significantly greater than that in vehicle control cultures and cultures containing AMH at 10 ng/ml, indicating the inhibition of growth initiation. Viability was unaffected by the presence of AMH when compared with tissues in control media. CONCLUSIONS: Recombinant AMH at a concentration of 100 ng/ml has an inhibitory effect on early human ovarian follicular development in vitro, suppressing the initiation of primordial follicle growth. PMID- 16720623 TI - Association of spermatogenic failure with decreased CDC25A expression in infertile men. AB - BACKGROUND: DAZ gene family is crucial for human spermatogenesis that requires the precise co-ordination of cell cycle events. CDC25A is recognized as the downstream substrate of DAZ gene family and is thought to function on the M-phase regulation of cell cycles. We investigated the expression profiles of CDC25A in the testes of infertile men and evaluated the relationship between CDC25A levels and testicular phenotype, clinical hormonal parameters and sperm retrieval results. METHODS: The protein and mRNA transcript levels of CDC25A in the testes of 40 azoospermic men were determined by immunohistochemistry and quantitative real-time-PCR. CDC25A in human spermatozoa was investigated by western blotting and immunofluorescence staining. RESULTS: The CDC25A protein was expressed mainly in spermatocyte, spermatid and spermatozoa. CDC25A transcript levels were significantly decreased (P = 0.0009) in patients with spermatogenic failure, especially in men with meiotic arrest and Sertoli cell-only syndrome. Significantly higher CDC25A transcript levels were detected in patients with successful sperm retrieval than in patients with failed sperm retrieval (P = 0.005). CONCLUSIONS: Decreased CDC25A is associated with spermatogenic failure and failed sperm retrieval in infertile men. Further studies are necessary to explore the functional roles of CDC25A in human spermatozoa. PMID- 16720624 TI - Progesterone receptor modulator CDB-2914 down-regulates vascular endothelial growth factor, adrenomedullin and their receptors and modulates progesterone receptor content in cultured human uterine leiomyoma cells. AB - BACKGROUND: This study was conducted to evaluate the effects of graded concentrations (10(-8), 10(-7) and 10(-6) M) of progesterone receptor (PR) modulator CDB-2914 on the protein contents of PR, of vascular endothelial growth factor (VEGF), adrenomedullin (ADM) and their receptors in cultured human uterine leiomyoma and matching myometrial cells. METHODS: PR-A, PR-B, VEGF-A, VEGF-B, VEGF receptor (VEGFR)-1, VEGFR-2, ADM and ADM receptor (ADMR) contents were assessed by Western blot analysis. RESULTS: Treatment with 100 ng/ml progesterone increased VEGF-A, VEGF-B and ADM contents in cultured leiomyoma cells and normal myometrial cells. The concomitant treatment with 10(-6) M CDB-2914 significantly decreased the progesterone-induced VEGF-A, VEGF-B and ADM contents in cultured leiomyoma cells but not in normal myometrial cells. CDB-2914 treatment alone decreased VEGFR-1, VEGFR-2 and ADMR contents in cultured leiomyoma cells but not in normal myometrial cells. CDB-2914 treatment increased PR-A and decreased PR-B contents in cultured leiomyoma cells in a dose-dependent manner compared with untreated cultures, whereas no significant changes in PR isoform contents were observed in normal myometrial cells. CONCLUSIONS: These results suggest that CDB 2914 down-regulates VEGF, ADM and their receptor contents and modulates PR isoform contents in cultured leiomyoma cells in a cell-type-specific manner. PMID- 16720625 TI - PGC-1alpha and PGC-1beta have both similar and distinct effects on myofiber switching toward an oxidative phenotype. AB - Peroxisome proliferator-activated receptor-gamma coactivator-1alpha and -1beta (PGC-1alpha and PGC-1beta) were overexpressed by adenovirus-mediated gene transfer in cultures of primary rat skeletal muscle cells derived from neonatal myoblasts. Effects on muscle fiber type transition and metabolism were studied from days 5 to 22 of culture. PGC-1alpha and PGC-1beta overexpression caused a three- to fourfold increase in mRNA level, a doubling of enzymatic activity of citrate synthase, a slight increase in short-chain acyl-CoA dehydrogenase mRNA, a doubling of the mRNA level, and a 30-50% increase in enzymatic activity of glyceraldehyde-3-phosphate dehydrogenase. Lactate dehydrogenase or creatine kinase activity was unchanged. PGC-1alpha enhanced glycogen buildup twofold at 5 or 25 mM glucose, whereas PGC-1beta caused a decrease. Both PGC-1alpha and PGC 1beta overexpression caused a faster maturation of myotubes, as seen by mRNA downregulation of the immature embryonal and perinatal myosin heavy-chain (MHC) isoforms. PGC-1alpha or PGC-1beta overexpression enhanced mRNA of the slow oxidative-associated MHC isoform MHCIb and downregulated mRNA levels of the fast glycolytic-associated MHC isoforms MHCIIX and MHCIIB. Only PGC-1beta overexpression caused an increase in mRNA of the intermediary fast oxidative associated MHC isoform MHCIIA. PGC-1alpha or PGC-1beta overexpression upregulated GLUT4 mRNA and downregulated myocyte enhancer factor 2C transcription factor mRNA; only PGC-1alpha overexpression caused an increase in the mRNA expression of TRB3, a negative regulator of insulin signaling. These results show that both PGC 1alpha and PGC-1beta are involved in the regulation of skeletal muscle fiber transition and metabolism and that they have both overlapping and differing effects. PMID- 16720626 TI - The regulation of IGFs and IGFBPs by prolactin in primary culture of fetal rat hepatocytes is influenced by maternal malnutrition. AB - During perinatal development, the regulation of IGF system appears to be growth hormone (GH) independent. By using highly purified primary fetal hepatocytes, we investigated the role of prolactin (PRL) in the regulation of IGF system and hepatocyte proliferation. We also analyzed the consequence of a maternal low protein (LP) diet on the regulation of IGF, IGF-binding protein (IGFBP), and hepatocyte proliferation by prolactin. Pregnant Wistar rats were fed a control (C) diet (20% protein) or isocaloric (LP; 8%) diet throughout gestation. On day 21.5, fetal hepatocytes were cultured for 4 days and incubated with rat prolactin. In the C hepatocytes, PRL at 100 ng/ml decreased the abundance of IGFBP-1 and IGFBP-2 by 50 (P < 0.05) and 60% (P < 0.01), respectively. It also reduced by 70% the level of IGF-II mRNA (P < 0.01). By contrast, PRL failed to modulate IGFBP-1 and IGFBP-2 production by LP hepatocytes, and this was associated with reduced abundance of the short form of PRL receptor (P < 0.05). PRL had no effect on either the proliferation or the IGF-I production by C and LP hepatocytes, although it reduced the expression of IGF-II. These results suggest that prolactin influences hepatocyte proliferation in vitro by inhibiting IGFBP 1, IGFBP-2, and IGF-II levels, which may coincide with the decline of IGF-II observed in rodents during late gestation in vivo. On the other hand, maternal LP diet induces a resistance of fetal hepatocytes to PRL. PMID- 16720627 TI - Assessment of postprandial glucose metabolism: conventional dual- vs. triple tracer method. AB - The dual-tracer method has been used conventionally for assessment of postprandial fluxes, i.e., appearance in plasma of ingested glucose (R(a meal)), endogenous glucose production (EGP), and disposal (R(d)). To quantify the magnitude of errors affecting the calculations and their dependence on model assumptions, this method was assessed and compared with the triple-tracer method, which provides model-independent estimates. For this purpose, the dual-tracer protocol was performed twice in eight normal subjects, with [1-(13)C]glucose to trace ingested glucose and [6,6-(2)H(2)]glucose constantly infused. A third tracer, [6-(3)H]glucose, was infused at variable rates to render the calculation of R(a meal) and EGP virtually model independent. The dual-tracer method analyzed with a one-compartment model performed poorly, since R(a meal) peak was significantly lower and delayed compared with triple-tracer reference, resulting in a significantly lower estimation of the amount of absorbed glucose (9,036 +/- 558 vs. 11,316 +/- 823 micromol/kg, P = 0.0117). EGP showed a paradoxical pattern, with an initial overshoot followed by a rapid decay to negative values, resulting in a significant underestimation of EGP suppression (57 +/- 3 vs. 65 +/ 4%, P = 0.0117). A two-compartment model performed better but did not overcome the limitations of the dual-tracer approach, since the amount of absorbed glucose was still significantly underestimated (10,231 +/- 661 vs. 12,169 +/- 838 micromol/kg, P = 0.0117) and EGP still showed a paradoxical behavior. R(d), estimated from R(a meal) and EGP, was significantly underestimated with the dual tracer method, irrespective of adopted model. We conclude that three suitably infused tracers are required for accurate assessment of postprandial R(a meal), EGP, and R(d). PMID- 16720628 TI - Exogenously imposed postprandial-like rises in systemic glucose and GLP-1 do not produce an incretin effect, suggesting an indirect mechanism of GLP-1 action. AB - The insulinotropic intestinal hormone GLP-1 is thought to exert one of its effects by direct action on the pancreatic beta-cell receptors. GLP-1 is rapidly degraded in plasma, such that only a small amount of the active form reaches the pancreas, making it questionable whether this amount is sufficient to produce a direct incretin effect. The aim of our study was to assess, in a dog model, the putative incretin action of GLP-1 acting directly on the beta-cell in the context of postprandial rises in GLP-1 and glucose. Conscious dogs were fed a high-fat, high-carbohydrate meal, and insulin response was measured. We also infused systemic glucose plus GLP-1, or glucose alone, to simulate the meal test values of these variables and measured insulin response. The results were as follows: during the meal, we measured a robust insulin response (52 +/- 9 to 136 +/- 14 pmol/l, P < 0.05 vs. basal) with increases in portal glucose and GLP-1 but only limited increases in systemic glucose (5.3 +/- 0.1 to 5.7 +/- 0.1 mmol/l, P = 0.1 vs. basal) and GLP-1 (6 +/- 0 to 9 +/- 1 pmol/l, P = 0.5 vs. basal). Exogenous infusion of systemic glucose and GLP-1 produced a moderate increase in insulin (43 +/- 5 to 84 +/- 15 pmol/l, 43% of the meal insulin). However, infusion of glucose alone, without GLP-1, produced a similar insulin response (37 +/- 6 to 82 +/- 14 pmol, 53% of the meal insulin, P = 0.7 vs. glucose and GLP-1 infusion). In conclusion, in dogs with postprandial rises in systemic glucose and GLP-1, the hormone might not have a direct insulinotropic effect and could regulate glycemia via indirect, portohepatic-initiated neural mechanisms. PMID- 16720629 TI - Recombinant human follicle-stimulating hormone alters maternal ovarian hormone concentrations and the uterus and perturbs fetal development in mice. AB - Gonadotropins are routinely administered to produce multiple oocytes for clinical in vitro fertilization (IVF) treatment, laboratory research, and livestock industries. Studies in mice have shown gonadotropin stimulation using equine chorionic gonadotropin (eCG) affects the endometrium, implantation, and fetal development. Evidence from clinical studies also indicates that stimulation with recombinant human follicle-stimulating hormone (rhFSH) may be detrimental to the endometrium and implantation rates. We investigated the effect of rhFSH in mice on maternal plasma hormone concentrations and uterine gene and protein expression and the effect of a stimulated maternal environment on pregnancy. Adult females were stimulated with rhFSH or eCG, followed by human chorionic gonadotropin (hCG). On day 4 of pseudopregnancy, mice either had embryos transferred to the uterus or were killed, and blood and uterine samples were collected. Pregnancy outcomes were examined on day 15. Gonadotropin stimulation increased plasma progesterone concentrations on day 4 compared with controls, whereas estradiol concentrations were unaffected. Stimulation also reduced uterine leukemia inhibitory factor (Lif) mRNA, but the expression of estrogen and progesterone receptors (Esr1 and Pgr), homeobox gene Hoxa10, and Vegf mRNA were unchanged. Furthermore, distribution of uterine PGR protein expression was altered by stimulation, but LIF protein was unchanged. Stimulated embryo transfer recipients had lower pregnancy rates than controls, and fetuses from the rhFSH group had reduced weight, length, and maturity. These results demonstrate that gonadotropin stimulation with rhFSH or eCG alters the preimplantation maternal environment, which results in reduced pregnancy rates and fetal development in the mouse. PMID- 16720630 TI - Maternal high-fat diet consumption results in fetal malprogramming predisposing to the onset of metabolic syndrome-like phenotype in adulthood. AB - Chronic consumption of a high-fat (HF) diet by female rats in their postweaning period resulted in significant increases in body weight and plasma levels of insulin, glucose, and triglycerides during pregnancy compared with female rats consuming a standard rodent laboratory chow (LC). On gestational day 21, plasma insulin levels and the insulin secretory response of islets to various secretogogues were significantly increased in HF fetuses. The HF male progeny weaned onto LC (HF/LC) demonstrated increases in body weight from postnatal day 60 onward. In adulthood, HF/LC male rats were significantly heavier than controls, had increased plasma levels of insulin, glucose, free fatty acids, and triglycerides, and demonstrated glucose intolerance. HF/LC male islets secreted increased amounts of insulin in response to low glucose concentrations, but their response to a high glucose concentration was similar to that of LC/LC islets. In another set of experiments, when the male progeny of HF female rats were weaned onto a high-sucrose diet (HF/HSu), their metabolic profile was further worsened. These results indicate that chronic consumption of a HF diet by female rats malprograms the male progeny for glucose intolerance and development of increased body weight in adulthood. The long-term high-fat feeding to female rats employed in this study bears resemblance to the dietary habits in Western societies. The results of this study implicate dietary practices of women in the etiology of the present epidemic of human obesity and related disorders. PMID- 16720631 TI - Metabolic effect of a LoBAG30 diet in men with type 2 diabetes. AB - We recently reported that in subjects with untreated type 2 diabetes a 5-wk diet of 30:30:40 carbohydrate/protein/fat ratio resulted in a significant decrease in 24-h integrated glucose, total %glycohemoglobin, and total cholesterol compared with a control diet of 55:15:30 carbohydrate/protein/fat given at the beginning of the 5-wk period. Body weight was stable and insulin was unchanged. We now present data on other hormones and metabolites considered to be affected by dietary macronutrient changes. The test diet resulted in an elevated fasting plasma total IGF-I, but not growth hormone. Urinary free cortisol was increased. Serum renin and urinary aldosterone remained unchanged. Blood pressure was stable. Serum creatinine and uric acid were increased. Urinary microalbumin was decreased. Creatinine clearance, serum B(12), folate, homocysteine, TSH, and free thyroxine were unchanged. Total triiodothyronine was decreased. Plasma alpha amino nitrogen, urea nitrogen, and serum albumin were increased. Urea production rate was increased such that a new steady state was present. The calculated urea production rate accounted for 84% of protein ingested on the control diet but only 68% on the test diet, suggesting net nitrogen retention on the latter. Overall, the lack of negative effects, the improved glucose control, and the positive nitrogen balance suggest such a diet will be beneficial for older subjects with type 2 diabetes. Nevertheless, the long-term effects and general applicability of the diet remain to be determined. PMID- 16720632 TI - Inbreeding depression and mixed mating in Leptosiphon jepsonii: a comparison of three populations. AB - BACKGROUND AND AIMS: Inbreeding depression is thought to play a central role in the evolution and maintenance of cross-fertilization. Theory indicates that inbreeding depression can be purged with self-fertilization, resulting in positive feedback for the selection of selfing. Variation among populations of Leptosiphon jepsonii in the timing and rate of self-fertilization provides an opportunity to study the evolution of inbreeding depression and mating systems. In addition, the hypothesis that differences in inbreeding depression for male and female fitness can stabilize mixed mating in L. jepsonii is tested. METHODS: In a growth room experiment, inbreeding depression was measured in three populations with mean outcrossing rates ranging from 0.06 to 0.69. The performance of selfed and outcrossed progeny is compared at five life history stages. To distinguish between self-incompatibility and early inbreeding depression, aborted seeds and unfertilized ovules were counted in selfed and outcrossed fruits. In one population, pollen and ovule production was quantified to estimate inbreeding depression for male and female fitness. KEY RESULTS: Both prezygotic barriers and inbreeding depression limited self seed set in the most outcrossing population. Cumulative inbreeding depression ranged from 0.297 to 0.501, with the lowest value found in the most selfing population. Significant inbreeding depression for early life stages was found only in the more outcrossing populations. Inbreeding depression was not significant for pollen or ovule production. CONCLUSIONS: The results provide modest support for the hypothesized relationship between inbreeding depression and mating systems. The absence of early inbreeding depression in the more selfing populations is consistent with theory on purging. Differences in male and female expression of inbreeding depression do not appear to stabilize mixed mating in L. jepsonii. The current estimates of inbreeding depression for L. jepsonii differ from those of previous studies, underscoring the effects of environmental variation on its expression. PMID- 16720633 TI - Periplasm turgor pressure controls wall deposition and assembly in growing Chara corallina cells. AB - BACKGROUND AND AIMS: New wall deposition usually accompanies plant growth. External osmotica inhibit both processes but wall precursors continue to be synthesized, and exocytosis follows. Consequently, the osmotica appear to act outside of the plasma membrane. Because this implies an action of turgor pressure (P) on the periplasm by unknown mechanisms, the following study was undertaken to determine whether P could act in a way that altered wall deposition and assembly in the periplasm while the cells grow. METHODS: Cells of Chara corallina were exposed to P slightly below normal by using a pressure probe while supplying inorganic carbon in light. After labelling, the walls were isolated and the amount of new wall was determined. Similar measurements were made after treatment with osmotica. Chlortetracycline-stimulated exocytosis was determined microscopically. Polysaccharide properties were determined by confocal microscopy and vapour pressure osmometry in an 'artificial periplasm' in isolated Chara cell walls, using labelled dextran as an analogue of hemicellulose, and polygalacturonate as pectin. KEY RESULTS: Rapid growth and wall deposition occurred at normal P of 0.5 MPa but both processes decreased when P was lowered 0.1 MPa. Inorganic carbon uptake and exocytosis were unaffected. In the artificial periplasm, normal P caused high polysaccharide concentrations and rapid polysaccharide entry into the wall, and gel formation in the pectin. Lowering P decreased entry and gel formation. CONCLUSIONS: This is the first indication that normal P of 0.5 MPa can concentrate periplasmic polysaccharides sufficiently to cause cross-linking and gel formation in pectins while simultaneously fostering the entry of large polysaccharides into small interstices in the existing wall. This P-action would thicken the primary wall and form a smooth transition between the new and old structure, suggesting a molecular mechanism of wall deposition and assembly while the wall extends. PMID- 16720634 TI - Etanercept treatment for three months is safe in patients with rheumatological manifestations associated with hepatitis C virus. AB - OBJECTIVE: The treatment of the rheumatological manifestations associated with hepatitis C virus (HCV) remains difficult. To examine the safety of anti-tumour necrosis factor-alpha treatment, nine patients having rheumatological manifestations associated with HCV were treated with etanercept 25 mg twice a week for 3 months. METHODS: Five patients had a positive viral load at study entry (Group I), four were negative (Group II). Clinical data recorded were: disease duration, painful and swollen joint count, patient global and physician global assessment, the number of 18 specified fibromyalgia tender points and the Health Assessment Questionnaire score. Laboratory studies included checking for the presence of cryoglobulinaemia and transaminase levels. Quantitative HCV viral RNA was performed by real-time polymerase chain reaction (PCR). RESULTS: At 3 months, no patient was found to have evidence of increased hepatic inflammation based on serial serum transaminase levels. In the five patients from Group I with detectable HCV RNA, no significant viral load increase was observed. No reactivation was observed in the four patients from Group II with undetectable HCV RNA. The effect on the clinical rheumatological manifestations was more heterogeneous but appears to be lower than that observed in rheumatoid arthritis. CONCLUSION: In this phase II open short-term study, etanercept appeared to be safe in patients with articular manifestations associated with HCV. PMID- 16720635 TI - Deficient activity of von Willebrand factor-cleaving protease in thrombotic thrombocytopenic purpura in the setting of adult-onset Still's disease. PMID- 16720636 TI - The -308 tumour necrosis factor-alpha gene polymorphism predicts therapeutic response to TNFalpha-blockers in rheumatoid arthritis and spondyloarthritis patients. AB - OBJECTIVE: To examine whether the G-to-A polymorphism at position -308 in the promoter of the tumour necrosis factor-alpha (TNFalpha) gene influences the therapeutic response to TNFalpha-blockers in patients with rheumatoid arthritis (RA), psoriatic arthritis (PsA) and ankylosing spondylitis (AS). METHODS: A total of 54 patients with RA, 10 with PsA and 22 with AS were genotyped by polymerase chain reaction for the -308 TNFalpha promoter polymorphism. They were treated with infliximab (n = 63), adalimumab (n = 10) or etanercept (n = 13). Clinical response was assessed after 24 weeks by the Disease Activity Score in 28 joints (DAS28) for RA and PsA, and the Bath Ankylosing Spondylitis Activity Index (BASDAI) for AS patients. RESULTS: All patients with the A/A genotype (n = 3, all RA) and two patients with the A/G genotype (AS) failed to respond to anti-TNF treatment. Irrespective of the underlying disease, moderate response (n = 44) was predominantly associated with the A/G genotype (A/G 18/22, G/G 4/22), whereas good response (n = 59) was exclusively seen in patients with the G/G genotype. The average improvement in the DAS28 score was 0.83 in the A/A, 1.50 in the A/G and 2.64 in the G/G group of RA and PsA patients (P < 0.0001). The BASDAI score in AS improved on average by 1.21 in the A/G and by 3.30 in the G/G group (P < 0.005). CONCLUSIONS: The data suggest that humans with a TNFalpha -308 G/G genotype are better responders to anti-TNFalpha treatment than those with A/A or A/G genotypes independent of the treated rheumatic disease (RA, PsA or AS). PMID- 16720637 TI - Towards a pro-inflammatory and immunomodulatory emerging role of leptin. AB - Leptin is a 16 kDa adipocyte-secreted hormone that regulates weight centrally and links nutritional status with neuroendocrine and immune function. Since its cloning in 1994, leptin's role in regulating immune and inflammatory response has become increasingly evident. Actually, the increase of leptin production that occurs during infection and inflammation strongly suggests that leptin is a part of the cytokines loop which governs the inflammatory-immune response and the host defence mechanism. Indeed, leptin stimulates the production of pro-inflammatory cytokines from cultured monocytes and enhances the production of Th1 type cytokines from stimulated lymphocytes. Several studies have implicated leptin in the pathogenesis of autoimmune inflammatory conditions such as type 1 diabetes, rheumatoid arthritis and chronic bowel disease. Obesity is characterized by elevated circulating leptin levels which might contribute significantly to the so called low-grade systemic inflammation, making obese individuals more susceptible to the increased risk of developing cardiovascular diseases, type II diabetes or inflammatory articular degenerative disease such as osteorathritis (OA). As a matter of fact, a key role for leptin in OA has been recently demonstrated since leptin exhibits, in synergy with other pro-inflammatory cytokines, a detrimental effect on articular cartilage cells by promoting nitric oxide synthesis. This review will focus prevalently on the complex relationships existing among leptin, inflammatory response and immunity, trying to provide surprising insights into leptin's role and to discuss challenges and prospects for the future. PMID- 16720638 TI - Cross-cultural adaptation, reliability and validity of the German Shoulder Pain and Disability Index (SPADI). AB - OBJECTIVE: To cross-culturally adapt the Shoulder Pain and Disability Index (SPADI) from English into German, and to test the reliability and validity of the German version. METHODS: Cross-cultural adaptation of the SPADI was performed according to international guidelines. One hundred and eighteen patients who had undergone shoulder arthroplasty, on average 4 yr previously, completed a questionnaire booklet containing the German SPADI, the Short Form 36 (SF-36), the Disability of the Arm, Shoulder and Hand (DASH) questionnaire, and the American Shoulder and Elbow Surgeons (ASES) questionnaire for the shoulder to assess SPADI's construct validity. One week later, they completed the SPADI again to assess test-retest reliability. RESULTS: The six-step cross-cultural adaptation procedure revealed no major problems with the content or language. The intraclass correlation coefficients for the individual items of the SPADI were between 0.68 and 0.89, and that for the SPADI total score was 0.94. The SPADI total score showed a correlation of 0.61-0.69 with the SF-36 physical scales, of 0.88 with the DASH and of 0.92 with the ASES. CONCLUSIONS: The German SPADI is a practicable, reliable and valid instrument, and can be recommended for the self assessment of shoulder pain and function. PMID- 16720639 TI - Upregulation of chicken p15INK4b at senescence and in the developing brain. AB - In mammalian cells, products of the INK4a-ARF locus play major roles in senescence and tumour suppression in different contexts, whereas the adjacent INK4b gene is more generally associated with transforming growth factor beta (TGF beta)-mediated growth arrest. As the chicken genome does not encode an equivalent of INK4a, we asked whether INK4b and/or ARF contribute to replicative senescence in chicken cells. In chicken embryo fibroblasts (CEFs), INK4b levels increase substantially at senescence and the gene is transcriptionally silenced in two spontaneously immortalised chicken cell lines. By contrast, ARF levels are unaffected by prolonged culture or immortalisation. These expression patterns resemble the behaviour of INK4a and ARF in human fibroblasts. However, short hairpin RNA (shRNA)-mediated knockdown of chicken INK4b or ARF provides only modest lifespan extension, suggesting that other factors contribute to senescence in CEFs. As well as underscoring the importance of the INK4b-ARF-INK4a locus in senescence, these findings imply that the encoded products have assumed different roles in different evolutionary niches. Although ARF RNA is not detectable in early chicken embryos, the INK4b transcript is expressed in the roof-plate of the developing hind-brain, consistent with a role in limiting cell proliferation. PMID- 16720640 TI - Human macrophages rescue myoblasts and myotubes from apoptosis through a set of adhesion molecular systems. AB - The mechanisms underlying stromal cell supportive functions are incompletely understood but probably implicate a mixture of cytokines, matrix components and cell adhesion molecules. Skeletal muscle uses recruited macrophages to support post-injury regeneration. We and others have previously shown that macrophages secrete mitogenic factors for myogenic cells. Here, we focused on macrophage elicited survival signals. We demonstrated that: (1) macrophage influx is temporally correlated with the disappearance of TUNEL-positive apoptotic myogenic cells during post-injury muscle regeneration in mice; (2) direct cell-cell contacts between human macrophages and myogenic cells rescue myogenic cells from apoptosis, as assessed by decreased annexin V labelling and caspase-3 activity, and by increased DIOC-6 staining, Bcl-2 expression and phosphorylation of Akt and ERK1/2 survival pathways; (3) four pro-survival cell-cell adhesion molecular systems detected by DNA macroarray are expressed by macrophages and myogenic cells in vitro and in vivo - VCAM-1-VLA-4, ICAM-1-LFA-1, PECAM-1-PECAM-1 and CX3CL1-CX3CR1; (4) macrophages deliver anti-apoptotic signals through all four adhesion systems, as assessed by functional analyses with blocking antibodies; and (5) macrophages more strongly rescue differentiated myotubes, which must achieve adhesion-induced stabilisation of their structure to survive. Macrophages could secure these cells until they establish final association with the matrix. PMID- 16720641 TI - Flat clathrin coats on endosomes mediate degradative protein sorting by scaffolding Hrs in dynamic microdomains. AB - Endocytosed membrane proteins that are destined for degradation in lysosomes are ubiquitylated and recognised by sorting complexes on endosome membranes. The ubiquitin-binding sorting component Hrs as well as ubiquitylated cargo are enriched in a characteristic flat clathrin coat on the endosome membrane. The function of clathrin within this coat has not been investigated. Here, we show that both clathrin and the clathrin-box motif of Hrs are required for the clustering of Hrs into restricted microdomains. The C-terminus of Hrs, which contains the clathrin-box, is sufficient to redirect a phosphatidylinositol(3) phosphate-binding protein into the Hrs- and clathrin-containing microdomains. Although these microdomains show little lateral diffusion in the membrane, they are dynamic structures that exchange Hrs and clathrin with similar kinetics, and acquire the downstream sorting component Tsg101. The clathrin-mediated clustering is essential for the function of Hrs in degradative protein sorting. We conclude that clathrin is responsible for concentrating Hrs in endosomal microdomains specialised for recognition of ubiquitylated membrane proteins, thus enabling efficient sorting of cargo into the degradative pathway. PMID- 16720642 TI - Mechanisms for human cytomegalovirus-induced cytoplasmic p53 sequestration in endothelial cells. AB - Human cytomegalovirus (HCMV) infection results in endothelial dysfunction, typically known as dysregulated apoptosis, and aberrant expression and sub cellular localization of p53, a tumor suppressor that accumulates at the late stage of infection. In this study, we examined three hypotheses that could be responsible for HCMV-induced cytoplasmic p53 accumulation at the later stage of infection: hyperactive nuclear export, cytoplasmic p53 tethering and delayed p53 degradation. Leptomycin B treatment, a nuclear export inhibitor, was unable to reduce cytoplasmic p53, thereby eliminating the hyperactive nuclear export mechanism. The findings that nascent p53 still entered nuclei after the nuclear export inhibition indicated that cytoplasmic tethering may play a minor role. Cytoplasmic p53 was still observed after the translation activities were blocked by cycloheximide. There was more than an eight-fold increase in the cytoplasmic p53 half-life with abnormal p53 ubiquitination. Taken together, these results suggest that delayed degradation could be responsible for the cytoplasmic p53 accumulation. The general slow-down of the proteasomal activity and the dysregulated p53 ubiquitination process at the later stage of infection could contribute to the reduced cytoplasmic p53 degradation and might be relevant to dysregulated endothelial apoptosis. The HCMV-induced changes in p53 dynamics could contribute to endothelial dysfunction. PMID- 16720644 TI - Palmitoylation determines the function of Vac8 at the yeast vacuole. AB - Palmitoylation stably anchors specific proteins to membranes, but may also have a direct effect on the function of a protein. The yeast protein Vac8 is required for efficient vacuole fusion, inheritance and cytosol-to-vacuole trafficking. It is anchored to vacuoles by an N-terminal myristoylation site and three palmitoylation sites, also known as the SH4 domain. Here, we address the role of Vac8 palmitoylation and show that the position and number of substrate cysteines within the SH4 domain determine the vacuole localization of Vac8: stable vacuole binding of Vac8 requires two cysteines within the N-terminus, regardless of the combination. Importantly, our data suggest that palmitoylation adds functionality to Vac8 beyond simple localization. A mutant Vac8 protein, in which the palmitoylation sites were replaced by a stretch of basic residues, still localizes to vacuole membranes and functions in cytosol-to-vacuole transport, but can only complement the function of Vac8 in morphology and inheritance if it also contains a single cysteine within the SH4 domain. Our data suggest that palmitoylation is not a mere hydrophobic anchor required solely for localization, but influences the protein function(s). PMID- 16720643 TI - Wingless signaling modulates cadherin-mediated cell adhesion in Drosophila imaginal disc cells. AB - Armadillo, the Drosophila homolog of beta-catenin, plays a crucial role in both the Wingless signal transduction pathway and cadherin-mediated cell-cell adhesion, raising the possibility that Wg signaling affects cell adhesion. Here, we use a tissue culture system that allows conditional activation of the Wingless signaling pathway and modulation of E-cadherin expression levels. We show that activation of the Wingless signaling pathway leads to the accumulation of hypophosphorylated Armadillo in the cytoplasm and in cellular processes, and to a concomitant reduction of membrane-associated Armadillo. Activation of the Wingless pathway causes a loss of E-cadherin from the cell surface, reduced cell adhesion and increased spreading of the cells on the substratum. After the initial loss of E-cadherin from the cell surface, E-cadherin gene expression is increased by Wingless. We suggest that Wingless signaling causes changes in Armadillo levels and subcellular localization that result in a transient reduction of cadherin-mediated cell adhesion, thus facilitating cell shape changes, division and movement of cells in epithelial tissues. PMID- 16720645 TI - Syndecan-1 regulates alphavbeta5 integrin activity in B82L fibroblasts. AB - B82L mouse fibroblasts respond to fibronectin or vitronectin via a syndecan-1 mediated activation of the alphavbeta5 integrin. Cells attached to syndecan-1 specific antibody display only filopodial extension. However, the syndecan anchored cells extend lamellipodia when the antibody-substratum is supplemented with serum, or low concentrations of adsorbed vitronectin or fibronectin, that are not sufficient to activate the integrin when plated alone. Integrin activation is blocked by treatment with (Arg-Gly-Asp)-containing peptides and function-blocking antibodies that target alphav integrins, as well as by siRNA mediated silencing of beta5 integrin expression. In addition, alphavbeta5 mediated cell attachment and spreading on high concentrations of vitronectin is blocked by competition with recombinant syndecan-1 ectodomain core protein and by downregulation of mouse syndecan-1 expression by mouse-specific siRNA. Taking advantage of the species-specificity of the siRNA, rescue experiments in which human syndecan-1 constructs are expressed trace the activation site to the syndecan-1 ectodomain. Moreover, both full-length mouse and human syndecan-1 co immunoprecipitate with the beta5 integrin subunit, but fail to do so if the syndecan is displaced by competition with soluble, recombinant syndecan-1 ectodomain. These results suggest that the ectodomain of the syndecan-1 core protein contains an active site that assembles into a complex with the alphavbeta5 integrin and regulates alphavbeta5 integrin activity. PMID- 16720647 TI - Abstracts of the 2nd North American Congress of Epidemiology, June 21-24, 2006, Seattle, Washington, USA. PMID- 16720648 TI - Distinct roles of cinnamate 4-hydroxylase genes in Populus. AB - Cinnamate 4-hydroxylase (C4H) catalyzes the conversion of cinnamate into 4 hydroxy-cinnamate, a key reaction of the phenylpropanoid pathway which leads to the biosynthesis of several secondary metabolites. C4H genes exist as a multigene family in various plant species. In order to understand the roles of individual C4H members, four C4H cDNAs (PtreC4H) were isolated from Populus tremuloides and three C4H loci (PtriC4H) were identified in the P. trichocarpa genome. The ability of Populus C4H isoforms to convert trans-cinnamate into p-coumaric acid was verified by the examination of yeast recombinant PtreC4H proteins. Populus C4H genes were expressed in various tissues, including developing xylem, phloem and epidermis; however, the expression patterns of individual members were different from each other. Sequential analysis of C4H promoters showed that the differential expression of C4H genes was associated with cis-acting regulatory elements such as box L, box P and H box, suggesting that the divergent C4H isoforms played distinct roles in the production of secondary metabolites. The involvement of specific C4H isoforms in the biosynthesis of guaiacyl and syringyl monolignols is discussed. PMID- 16720646 TI - Basal body and flagellum mutants reveal a rotational constraint of the central pair microtubules in the axonemes of trypanosomes. AB - Productive beating of eukaryotic flagella and cilia requires a strict regulation of axonemal dynein activation. Fundamental to any description of axonemal beating is an understanding of the significance of the central pair microtubules and the degree to which central pair rotation has a role. However, for the majority of organisms, it is unclear whether the central pair actually rotates. Using an extra-axonemal structure as a fixed reference, we analysed the orientation of the central pair in African trypanosomes and other kinetoplastid protozoa. A geometric correction allowed the superposition of data from many cross-sections, demonstrating that the axis of the central pair is invariant and that there is no central pair rotation in these organisms. Analysis of mutants depleted in particular flagellar and basal body proteins [gamma-tubulin, delta-tubulin, Parkin co-regulated gene product (PACRG) or the paraflagellar rod protein PFR2] allowed a dissection of the mechanisms for central pair constraint. This demonstrated that orientation is independent of flagellum attachment and beating, but is influenced by constraints along its length and is entirely dependent on correct positioning at the basal plate. PMID- 16720649 TI - ACT domain repeat protein 7, ACR7, interacts with a chaperone HSP18.0-CII in rice nuclei. AB - The regulatory ACT domains serve as amino acid-binding sites in some amino acid metabolic enzymes and transcriptional regulators in bacteria. To elucidate the molecular roles of the glutamine (Gln)-sensing system in nitrogen (N) metabolism in plants, we isolated six genes encoding ACT domain repeat proteins (ACR1, and ACR5-ACR9) from rice (Oryza sativa L.) using genomic information on the primary structure composed of four copies of the domain homologous to those of bacterial Gln sensor GLND. Since expression of ACR7 was the most abundant of the six ACR orthologous genes, we focused on this ACR in the current study. Gene products of ACR7 were most abundant in young developing leaf blades of rice, and ACR7 protein is specifically localized in the nucleus of the parenchyma cells of phloem and xylem in the vascular bundles. A yeast two-hybrid screen identified a small heat stress protein (HSP18.0-CII) as a protein interacting with ACR7. Transient expression analysis of HSP18.0-CII:sGFP in cultured rice cells, followed by co immunoprecipitation, suggests that the nuclear ACR7 indeed interacted with nucleocytoplasmic HSP18.0-CII in vivo. The potential ability of nuclear protein ACR7 to bind Gln and the possibility of the protein acting as a Gln sensor in rice leaves is discussed. PMID- 16720650 TI - C-3 epimers can account for a significant proportion of total circulating 25 hydroxyvitamin D in infants, complicating accurate measurement and interpretation of vitamin D status. AB - CONTEXT: We have recently introduced liquid chromatography-tandem mass spectrometry (LC-MS/MS) for 25-hydroxyvitamin D(2) (25OHD(2)) and 25OHD(3) testing. During subsequent clinical use, we identified significantly elevated results in some infants. We hypothesized this might represent assay interference caused by C-3 epimers of 25OHD(2) or 25OHD(3). OBJECTIVE: Our aims were to 1) determine the prevalence of C-3 epimers of 25OHD(2) or 25OHD(3) in human serum, and 2) identify the patient populations that might be affected. STUDY DESIGN: We modified our LC-MS/MS method to allow detection of C-3 epimers. We retested specimens from four patient groups with the new method and an extracted RIA: 1) children less than 1 yr old, 2) children 1-18 yr old, 3) adults aged 20-87 yr with liver disease, and 4) adults aged 19-91 yr without liver disease. RESULTS: In 172 children from group 1 with detectable 25OHD(2) or 25OHD(3), we identified C-3 epimers in 39 (22.7%). The epimers contributed 8.7-61.1% of the total 25-OHD. There was an inverse relationship between patient age and epimer percentage (r = 0.48; P < 0.002). The RIA gave accurate 25-OHD results that correlated with the modified LC-MS/MS method. No C-3 epimers were detected in any of the other groups. CONCLUSIONS: Significant concentrations of C-3 epimers of 25OHD(2) or 25OHD(3) are commonly found in infants. This can lead to overestimation of 25-OHD levels. Measurements in children less than 1 yr should therefore be performed with an assay that allows accurate detection of 25-OHD in the presence of its C-3 epimers. PMID- 16720651 TI - The response of luteinizing hormone pulsatility to 5 days of low energy availability disappears by 14 years of gynecological age. AB - CONTEXT: The incidence of menstrual disorders declines during adolescence. The mechanism responsible is unknown. OBJECTIVE: The objective of the study was to test the hypothesis, formulated a priori, that the dependence of LH pulsatility and ovarian function on energy availability declines with gynecological age (years since menarche). DESIGN: The study was a controlled experiment repeated in two menstrual cycles, performed 2001-2004. SETTING: The study was conducted at a university laboratory and general clinical research center. PARTICIPANTS: The study population consisted of healthy, habitually sedentary, young women of normal body composition with 5-8 yr (adolescents, n = 9) and 14-18 yr (adults, n = 10) of gynecological age recruited by advertisement from approximately 9000 women aged 18-34 yr in a college community. Samples were similar in age of menarche, length of menstrual cycle and luteal phase, body size and composition, aerobic capacity, and dietary intake. None were withdrawn due to adverse effects. INTERVENTIONS: Interventions included energy availabilities of 45 and 10 kcal/kg of fat-free mass per day for 5 d in the early follicular phases of separate menstrual cycles in random order. MAIN OUTCOME MEASURES: LH pulsatility, estradiol, and luteal phase length were measured. RESULTS: Low energy availability reduced LH pulse frequency in adolescents (P < 0.01) but not adults (P = 0.39), did not increase LH pulse amplitude in either group (both P = 0.13), and suppressed 24-h mean LH in adolescents (P = 0.01) but not adults (P = 0.72). Estradiol was unaffected (both P = 0.48), but the subsequent luteal phase was shorter in adolescents (P < 0.01). CONCLUSIONS: In women of normal body composition, the response of LH pulsatility and ovarian function to 5 d of low energy availability disappears by 14 yr of gynecological age. PMID- 16720652 TI - Comparative pharmacokinetics and pharmacodynamics of a new sustained-release growth hormone (GH), LB03002, versus daily GH in adults with GH deficiency. AB - CONTEXT: LB03002 is a novel sustained-release GH preparation administered once weekly. OBJECTIVE: Our objective was to examine the pharmacokinetics, pharmacodynamics, and safety of LB03002 vs. daily GH. DESIGN AND SETTING: This open-label, crossover study compared the pharmacokinetics and pharmacodynamics of LB03002 and daily GH. PATIENTS AND OTHER PARTICIPANTS: Six male and three female patients with adult GH deficiency participated in the single-center study. INTERVENTION: Subjects were on stable daily GH treatment before the study. After a 4-wk washout with no GH, five weekly doses of LB03002 were given. MAIN OUTCOME MEASURE: GH and IGF-I concentrations were measured during the last dose of daily GH and during the first and fifth weekly doses of LB03002. RESULTS: The observed maximal serum GH concentration was approximately doubled after LB03002 (6.1 +/- 3.2 and 4.5 +/- 2.2 microg/liter at first and fifth doses) compared with daily GH (2.7 +/- 2.2 microg/liter). A sustained increase in GH concentration for more than 48 h was observed with LB03002, such that dose-normalized area under the curve (AUC) was not significantly different between daily GH and LB03002. Mean maximal serum IGF-I concentration was 34-41% greater with LB03002 than with daily GH, and AUC was 7-fold greater. However, normalized to GH dose, AUC for IGF-I was comparable. Adverse events and local reactions were acceptable, and there were no evident safety concerns with LB03002. CONCLUSIONS: Multiple weekly doses of LB03002 appeared safe and well tolerated. Comparable GH bioavailability and sustained IGF-I elevations support the use of once-weekly LB03002 to replace daily GH therapy. PMID- 16720653 TI - Comparison of follicle-stimulating-hormone-stimulated dimeric inhibin and estradiol responses as indicators of granulosa cell function in polycystic ovary syndrome and normal women. AB - CONTEXT: Follicular phase secretion of inhibin B, like that of estradiol (E(2)), correlates with the quantity and quality of developing follicles. However, it has not been established whether inhibin B responses to gonadotropin stimulation parallel those of E(2) as a reflection of granulosa cell functional capacity. OBJECTIVE: Our objective was to determine whether inhibin B responses to FSH stimulation are similar to those of E(2) in women with polycystic ovary syndrome (PCOS) and normal women. DESIGN AND SETTING: We conducted a prospective study to compare ovarian responses in two groups of women at a general clinical research center in a tertiary academic medical center. PATIENTS: Women with PCOS, 18-35 yr (n = 19), and normal ovulatory controls, 18-35 yr (n = 7), were recruited for study. INTERVENTIONS: Serum samples were measured over a 24-h period after an iv injection of recombinant human FSH, 150 IU. MAIN OUTCOME MEASURES: Serum E(2), inhibin A, and inhibin B responses after FSH administration were assessed. RESULTS: In PCOS women, the 24-h production of inhibin B and E(2) after FSH was significantly greater than that of normal controls. Within the PCOS group, the fold change in inhibin B was significantly greater than that of E(2). Inhibin A responses between groups were similar and of markedly lower magnitude. CONCLUSIONS: FSH-stimulated inhibin B responses may be employed to assess the functional capacity of granulosa cells in PCOS and normal women. PMID- 16720654 TI - Reduced plasma visfatin/pre-B cell colony-enhancing factor in obesity is not related to insulin resistance in humans. AB - CONTEXT: Visfatin was recently identified as a protein highly expressed and secreted in adipose tissue with insulin-mimetic effect and is a candidate hormone to help explain the association among adipose tissue expansion, insulin resistance, and type 2 diabetes. OBJECTIVE: The objective of the study was to assess expression of visfatin in lean and obese subjects and in sc and visceral adipose tissue and moreover to explore the role of visfatin on insulin resistance in humans. DESIGN: We measured circulating visfatin and its mRNA expression in sc adipose tissue (SAT) in lean and obese subjects. Furthermore, we measured visfatin mRNA in visceral adipose (VAT) and SAT by quantitative RT-PCR. Finally, plasma visfatin and its mRNA in SAT were measured under free fatty acid-induced insulin resistance in healthy subjects. RESULTS: Plasma visfatin and its mRNA in SAT were significantly lower in obese subjects, compared with normal-weight controls. Both circulating visfatin and SAT visfatin mRNA were negatively correlated with body mass index, whereas no correlation was found with homeostasis model assessment. Significantly higher visfatin mRNA was found in VAT of obese subjects, compared with lean controls. Interestingly, visfatin mRNA in VAT was positively correlated with BMI. Elevation of free fatty acid induced a condition of insulin resistance but did not affect either circulating visfatin or its mRNA. CONCLUSIONS: Our findings show that, in human obesity, plasma visfatin is reduced, whereas visfatin mRNA is differentially regulated in SAT and VAT. Visfatin is not related to insulin resistance either as assessed by homeostasis model assessment or during lipid infusion. PMID- 16720655 TI - Effect of long-term calorie restriction with adequate protein and micronutrients on thyroid hormones. AB - CONTEXT: Caloric restriction (CR) retards aging in mammals. It has been hypothesized that a reduction in T(3) hormone may increase life span by conserving energy and reducing free-radical production. OBJECTIVE: The objective of the study was to assess the relationship between long-term CR with adequate protein and micronutrient intake on thyroid function in healthy lean weight stable adult men and women. DESIGN, SETTING, AND PARTICIPANTS: In this study, serum thyroid hormones were evaluated in 28 men and women (mean age, 52 +/- 12 yr) consuming a CR diet for 3-15 yr (6 +/- 3 yr), 28 age- and sex-matched sedentary (WD), and 28 body fat-matched exercising (EX) subjects who were eating Western diets. MAIN OUTCOME MEASURES: Serum total and free T(4), total and free T(3), reverse T(3), and TSH concentrations were the main outcome measures. RESULTS: Energy intake was lower in the CR group (1779 +/- 355 kcal/d) than the WD (2433 +/- 502 kcal/d) and EX (2811 +/- 711 kcal/d) groups (P < 0.001). Serum T(3) concentration was lower in the CR group than the WD and EX groups (73.6 +/- 22 vs. 91.0 +/- 13 vs. 94.3 +/- 17 ng/dl, respectively) (P < or = 0.001), whereas serum total and free T(4), reverse T(3), and TSH concentrations were similar among groups. CONCLUSIONS: Long-term CR with adequate protein and micronutrient intake in lean and weight-stable healthy humans is associated with a sustained reduction in serum T(3) concentration, similar to that found in CR rodents and monkeys. This effect is likely due to CR itself, rather than to a decrease in body fat mass, and could be involved in slowing the rate of aging. PMID- 16720656 TI - The value of follicle-stimulating hormone concentration and clinical findings as markers of the late menopausal transition. AB - CONTEXT: The Stages of Reproductive Aging Workshop proposed bleeding and hormonal criteria for the menopausal transition, but operational definitions of hormone parameters were not specified. OBJECTIVE: This paper investigates the longitudinal relationship of annual serum FSH levels with four proposed bleeding criteria for the late menopausal transition in two cohort studies. The goal is to provide empirically based guidance regarding application of hormonal criteria that may be optimal for widespread application in clinical and research settings for assessing menopausal stage. DESIGN/SETTING: Prospective menstrual calendar and annual serum FSH data were collected from two population-based cohort studies: the Melbourne Women's Midlife Health Project and the Study of Women's Health Across the Nation. PARTICIPANTS: Participants in the study were 193 Melbourne Women's Midlife Health Project and 2223 Study of Women's Health Across the Nation women aged 42-57 yr at baseline who contributed 10 or more menstrual cycles and at least one annual serum FSH value. MAIN OUTCOME MEASURE(S): Association between bleeding criteria for the late menopausal transition and FSH was a main outcome measure. Associations of bleeding criteria, FSH, and hot flashes with the final menstrual period were also measured. RESULTS: A single FSH measure is an independent marker of the late menopausal transition, but FSH concentrations are less predictive of menopausal stage than any of four proposed bleeding criteria. Criterion FSH values for the late transition are similar across both studies. Experience of hot flashes adds no information in the presence of hormonal and bleeding criteria. CONCLUSIONS: An annual serum FSH concentration of 40 IU/liter could be incorporated, in conjunction with bleeding markers, into the Stages of Reproductive Aging Workshop paradigm for markers of the late menopausal transition. PMID- 16720657 TI - Serum profiles of free and conjugated neuroactive pregnanolone isomers in nonpregnant women of fertile age. AB - BACKGROUND: Pregnanolone isomers (PI) with a hydroxy group in the 3alpha-position are neuroinhibitors operating via positive modulation of GABA(A) receptors. The 3beta-PI and sulfates of PI and pregnenolone exert the opposite effect. In addition to the brain's in situ synthesis, some circulating steroids can penetrate the blood-brain barrier. METHODS: To assess the physiological impact of peripheral endogenous neuroactive pregnanolone isomers and their polar conjugates in women, serum allopregnanolone (P3alpha5alpha), isopregnanolone (P3beta5alpha), pregnanolone (P3alpha5beta), epipregnanolone (P3beta5beta), pregnenolone, estradiol (including their polar conjugates), and additional steroids were measured in 16 women in the follicular and luteal phases of the menstrual cycle using gas chromatography/mass spectrometry and RIA for the analysis. Linear models and Spearman's correlations were used for data evaluation. RESULTS AND DISCUSSION: The levels of conjugated PI were from one to almost three orders of magnitude higher in comparison with the free steroids. The results indicate that a substantial proportion of the progesterone is metabolized in the sequence progesterone-->5beta-dihydroprogesterone-->P3alpha5beta-->conjugated P3alpha5beta. The sulfation of PI and particularly of P3alpha5beta moderates the levels of free PI and restrains estradiol biosynthesis via progesterone degradation. PI including the conjugates reflected changing progesterone formation during the menstrual cycle. In the follicular phase, the positive correlation with conjugated pregnenolone, the independence of progesterone, and the negative age relationships of PI indicate their adrenal origin. The dependence on progesterone and the independence of conjugated pregnenolone suggest a gonadal source of PI in the luteal phase. The neuroactivating PI prevailed over neuroinhibiting PI. PMID- 16720658 TI - Haplotype analysis reveals founder effects of thyroglobulin gene mutations C1058R and C1977S in Japan. AB - CONTEXT: Thyroglobulin (Tg) mutations were previously believed to be rare, resulting in congenital goitrous hypothyroidism. However, an increasing number of patients with Tg mutations, who are euthyroid to mildly hypothyroid, have been identified in Japan. OBJECTIVES: The purpose of this study was to investigate whether the three frequently found Tg mutations, namely C1058R, C1245R, and C1977S, were caused by a founder effect. RESULTS: We found 26 different mutations within the Tg gene in 52 patients from 41 families. Thirty-five patients were homozygous for the mutations, whereas the others were compound heterozygous. The occurrence of Tg mutation within the general Japanese population is one in 67,000. Patients with the C1245R mutation were found throughout Japan, whereas those with the C1058R mutation were confined to a small village on a southern island, and those with the C1977S mutation were restricted to a city. The eight patients with the C1058R mutation and the seven patients with the C1977S mutation all showed the same combinations of 18 single-nucleotide polymorphisms in the coding region of the Tg gene, which would appear in one in 810 million and one in 37 billion, respectively, control subjects. CONCLUSIONS: The frequently found mutations, C1058R and C1977S, were caused by founder effects. This result suggests that Tg mutations may provide a genetic basis for the cause of familial euthyroid goiter. PMID- 16720659 TI - Increased anti-Mullerian hormone serum concentrations in prepubertal daughters of women with polycystic ovary syndrome. AB - CONTEXT: Anti-Mullerian hormone (AMH) is produced by the granulosa cells and reflects follicular development. Adult women with polycystic ovary syndrome (PCOS) have increased levels of AMH associated with an excessive number of growing follicles. However, it is not known whether these abnormalities are present before the clinical onset of PCOS. OBJECTIVE: Our objective was to investigate whether prepubertal daughters of women with PCOS have increased AMH levels. DESIGN: Fourteen female infants (2-3 months old) and 25 prepubertal girls (4-7 yr old) born to PCOS mothers were studied. As a control group, we studied 21 female infants and 24 prepubertal girls born to mothers with regular menses and without hyperandrogenism. The group with PCOS mothers and the control group had normal birth weight and were born from spontaneous singleton pregnancies. Circulating concentrations of gonadotropins, testosterone, androstenedione, estradiol, 17-OH-progesterone, SHBG, inhibin B, and AMH were determined by specific assays. RESULTS: Serum concentrations of AMH were significantly higher in the PCOS group compared with the control group during early infancy (20.4 +/- 15.6 vs. 9.16 +/- 8.6 pmol/liter; P = 0.024) and during childhood (14.8 +/- 7.7 vs. 9.61 +/- 4.4 pmol/liter; P = 0.007). Gonadotropin and serum sex steroid concentrations were similar in both groups during the two study periods, except for FSH, which was lower during childhood in girls born to PCOS mothers. CONCLUSIONS: We conclude that serum AMH concentrations are increased in prepubertal daughters of PCOS women, suggesting that these girls appear to show evidence of an altered follicular development during infancy and childhood. PMID- 16720660 TI - The prevalence of the 65-kilodalton isoform of glutamic acid decarboxylase autoantibodies by glucose tolerance status in elderly patients from the cardiovascular health study. AB - CONTEXT: Autoantibodies (AA) to glutamic acid decarboxylase (GAD65), a determinant of risk for autoimmune diabetes, have been found in up to 10% of patients with type 2 diabetes. In older adults, this marker may also serve as a determinant of risk for autoimmune diabetes and enhance diabetes classification. OBJECTIVE: The objective of this study was to evaluate the relationship between GAD65AA and glucose tolerance status, current diabetes treatment, and clinical measures in older adults. DESIGN: GAD65AA were measured at baseline in 3318 participants from the Cardiovascular Health Study, a cohort study of 5888 individuals 65 or older. SETTING: The population-based cohort was recruited from four U.S. sites. PATIENTS: Patients included all Cardiovascular Health Study participants with known diabetes, newly diagnosed diabetes, impaired fasting glucose, impaired glucose tolerance, and a sample of normal glucose-tolerant participants. MAIN OUTCOME MEASURES: GAD65AA, body mass index, fasting glucose and insulin levels, blood pressure, lipid levels, and diabetes treatment at baseline were measured. RESULTS: The prevalence of GAD65AA increased with decreasing glucose tolerance in both Blacks (n = 560) and Whites (n = 2730), being more pronounced in known diabetic individuals. GAD65AA were found in 2.3, 5.8, 7.8, and 8.3% of diabetic participants, reporting use of no diabetes medication, oral hypoglycemic agents, insulin only, and both oral hypoglycemic agents and insulin, respectively (P = 0.02, linear trend). Among diabetic participants, GAD65AA positivity was associated with diabetes treatment, higher fasting glucose, and lower body mass index. CONCLUSIONS: Even among older individuals with diabetes, GAD65AA may be a useful marker in identifying a subgroup of autoimmune diabetes, serve as a marker of insulin requirement, and remain stable over years. PMID- 16720661 TI - Pegvisomant for the treatment of gsp-mediated growth hormone excess in patients with McCune-Albright syndrome. AB - CONTEXT: GH excess affects approximately 20% of the patients with McCune-Albright syndrome (MAS). MAS is caused by sporadic, postzygotic, activating mutations in the GNAS gene, which codes for the cAMP-regulating protein, G(s)alpha (gsp oncogene). These same mutations are found in approximately one third of the sporadic cases of acromegaly. OBJECTIVE: We examined efficacy of the GH receptor antagonist, pegvisomant, in controlling gsp oncogene-mediated GH excess and skeletal disease (fibrous dysplasia of bone) associated with MAS. SETTING AND PATIENTS: Five MAS patients with GH excess were treated with 20 mg/d sc injection of pegvisomant for 12 wk in a randomized, double-blind, placebo-controlled crossover study at the National Institutes of Health. MAIN OUTCOME MEASURES: The primary measure of efficacy was normalization of IGF-I. Secondary outcome measures were reduction in serum IGF binding protein-3 (IGFBP-3), improvement of fatigue and sweating, and reduction in markers of bone metabolism and bone pain. RESULTS: Combined mean changes in serum IGF-I at 6 and 12 wk were -236.4 ng/ml (53%, P < 0.005) and -329.8 ng/ml (62%, P < 0.001), respectively. IGFBP-3 decreased by 0.8 mg/liter (24%, P < 0.01) and 2.9 mg/liter (37%, P < 0.005), respectively. There were no significant changes in signs and symptoms of acromegaly or markers of bone metabolism and bone pain, nor was there a significant change in pituitary size. Retrospective comparison of the degree of control achieved with pegvisomant vs. other medications (long-acting octreotide +/- dopamine agonist) in the same group showed that the two regimens were similarly effective. CONCLUSIONS: Pegvisomant effectively reduced IGF-I and IGFBP 3 levels in gsp-mediated GH excess but had no effect on fibrous dysplasia. PMID- 16720662 TI - Self-esteem and social adjustment in young women with Turner syndrome--influence of pubertal management and sexuality: population-based cohort study. AB - CONTEXT: Pediatric management of patients with Turner syndrome focuses on height, frequently resulting in a delay of pubertal induction. The influence of pubertal management on psychosocial adjustment and sex life has not been evaluated in Turner syndrome patients. OBJECTIVE: The objective of the study was to identify the determinants of self-esteem, social adjustment, and initiation of sex life in patients with Turner syndrome, particularly those related to pubertal management. DESIGN: This was a prospective evaluation, the StaTur study. SETTING: The study was conducted with a population-based registry of GH-treated patients. PARTICIPANTS: Participants included 566 young adult women with Turner syndrome, aged 22.6 +/- 2.6 yr (range, 18.3-31.2). MAIN OUTCOME MEASURES: Measures used in the study were Coopersmith's Self-Esteem Inventory, Social Adjustment Scale Self Report, questions on sexual experience, and extensive data on pediatric management. RESULTS: Low self-esteem was associated with otological involvement and limited sexual experience. Low social adjustment was associated with lower paternal socioeconomic class and an absence of sexual experience. Late age at first kiss or date was associated with cardiac involvement and a lack of spontaneous pubertal development. Age at first sexual intercourse was related to age at puberty and paternal socioeconomic class. Delayed induction of puberty had a long-lasting effect on sex life. Height and height gain due to GH treatment had no effect on outcomes. CONCLUSIONS: Puberty should be induced at a physiologically appropriate age in patients with Turner syndrome to optimize self esteem, social adjustment, and initiation of the patient's sex life. Therapeutic interventions altering normal pubertal development in other groups of patients should be reconsidered in light of these findings. PMID- 16720663 TI - Syndromic obesity and diabetes: changes in body composition with age and mutation analysis of ALMS1 in 12 United Kingdom kindreds with Alstrom syndrome. AB - CONTEXT: Alstrom syndrome (AS) is a monogenic form of infancy-onset obesity and insulin resistance, caused by ALMS1 mutations. The natural history of the insulin resistance is unknown, in particular how this relates to changes in body composition. It is also unclear how ALMS1 mutations relate to the characteristic phenotype. OBJECTIVES: Our objectives were to characterize body composition and metabolic parameters, to establish ALMS1 mutation spectrum of United Kingdom AS patients, and to determine whether a genotype-phenotype correlation exists. DESIGN AND PATIENTS: We conducted a cross-sectional cohort study of 12 unrelated subjects with AS. Age-standardized body composition was assessed by anthropometry and dual-energy x-ray absorptiometry and insulin sensitivity by homeostasis model assessment. The exons and intron-exon boundaries of ALMS1 were directly sequenced. SETTING: The study was performed during the annual Alstrom Syndrome UK multidisciplinary screening clinic. RESULTS: AS patients have early-onset obesity, but body mass index, waist circumference, and body fat from dual-energy x-ray absorptiometry were negatively correlated with age (r = -0.37, P = 0.2; r = -0.84, P = 0.002; and r = -0.6, P = 0.05). Despite this, insulin resistance increased, demonstrated by raised fasting insulin and fall in homeostasis model assessment insulin sensitivity with age (r = -0.64, P = 0.02). ALMS1 mutations were identified in 10 of 12 patients, with a potential founder mutation in exon 16 present in five [np 10775del (C); Del3592fs/ter3597]. No genotype-phenotype correlation was observed. CONCLUSIONS: We identified mutations in ALMS1 in more than 80% of patients with no genotype-phenotype correlation. In AS, severe childhood obesity, waist circumference, and body fat decrease with age, whereas insulin resistance increases. The abdominal obesity, insulin resistance, diabetes, hypertriglyceridemia, and hypertension suggest that AS could represent a monogenic model for the metabolic syndrome. PMID- 16720664 TI - Ghrelin and peptide YY in youth: are there race-related differences? AB - OBJECTIVE: Obesity prevalence is higher in African-American (AA) vs. American white (AW) youth. Ghrelin is a "hunger" peptide that is high preprandially and decreases postprandially, and peptide YY (PYY) is a "satiety" hormone increasing after meals. Impaired regulation of ghrelin/PYY may be conducive to obesity. We hypothesized that racial differences in childhood obesity could partly be explained by differences in ghrelin/PYY dynamics. RESEARCH DESIGN AND METHODS: We investigated: 1) ghrelin suppression/PYY elevation in response to an oral glucose tolerance test (OGTT) in AA vs. AW, and 2) the relationship of ghrelin and PYY dynamics to insulin sensitivity. Thirty-three AA and 54 AW prepubertal children underwent an OGTT measuring ghrelin, PYY, glucose, and insulin. Fasting glucose to insulin ratio (G(F)/I(F)) was used to assess the relationship of insulin sensitivity to fasting and post-OGTT ghrelin and PYY levels. RESULTS: OGTT induced suppression in ghrelin (Delta ghrelin) was lower in AA youth. Delta ghrelin correlated with G(F)/I(F) (r = 0.47, P < 0.001) and Delta insulin at 30 min (r = -0.47, P < 0.001). In multiple regression analysis, race (P = 0.013) and G(F)/I(F) (P = 0.004) contributed independently to the variance in Delta ghrelin (R(2) = 0.28, P < 0.001). Fasting and post-OGTT PYY levels were lower in AAs and were not related to insulin sensitivity. CONCLUSION: The lower suppression of ghrelin in AA, but not the lower PYY levels, correlates with insulinemia and insulin resistance. Less ghrelin suppression and PYY elevation after a meal in black youth could be a potential mechanism of race-related differences in hunger/satiety predisposing to risk of obesity. PMID- 16720665 TI - Laparoscopic adrenalectomy for potentially malignant adrenal tumors greater than 5 centimeters. AB - PURPOSE: Laparoscopic adrenalectomy (LA) is controversial for large, potentially malignant tumors. We report a series of LA or hand-assisted LA for large (>5 cm) adrenal tumors. PATIENTS AND METHODS: Among 210 LAs performed in 6 yr, 39 patients had potentially malignant tumors greater than 5 cm in diameter. Their perioperative and follow-up data were retrospectively analyzed. RESULTS: All 39 patients had successful LAs without perioperative mortality, conversion to open surgery, or capsular disruption during dissection. The mean tumor size was 6.2 cm (range, 5-12 cm), operative time 207 min (115-315 min), and blood loss 75 ml (minimal-1400 ml). Complications included one intraoperative diaphragmatic perforation, three mild wound infections, and one pneumonia. Preoperatively there were 27 nonfunctioning tumors, seven pheochromocytomas, three cortisol-secreting tumors, and two virilizing tumors. Final pathology revealed eight malignant (four adrenocortical carcinomas and four metastatic carcinomas) and 31 benign tumors (14 cortical adenomas, eight pheochromocytomas, six myelolipomas, and three ganglioneuromas). Median follow-up was 39 months. Four patients (two adrenocortical carcinomas, one metastatic hepatoma, and one lymphoma) died 24, 10, 9, and 3 months after surgery, respectively. A hand-assisted device was used in 10 patients. Only the tumor size was larger and length of postoperative hospital stay longer for those in the hand-assisted group. CONCLUSIONS: LA is a reasonable option for selected large adrenal tumors when complete resection is technically feasible and there is no evidence of local invasion. Hand-assisted LA is a good alternative to open conversion if a difficult dissection is encountered intraoperatively. PMID- 16720666 TI - Timing of puberty determines serum insulin-like growth factor-I in late adulthood. AB - CONTEXT: IGFs may play an important role in disease etiology, especially cancer. Changes in diet can alter acute levels, but little is known about life course influences on IGF levels. OBJECTIVE: The objective of the study was to examine the association between timing of puberty and adulthood serum IGFs (IGF-I and IGF binding protein-3). DESIGN: This was a retrospective cohort study. SETTING: Male pupils who attended a single school in Southern England were part of the study. PARTICIPANTS: Participants in the study were a cohort of 1028 men born between 1927 and 1956 with anthropometric measures between 9 and 18 yr and adulthood serum IGF levels. MAIN OUTCOME MEASURE: The study measured serum IGF-I and IGF binding protein-3 at mean age 63 yr. RESULTS: Age at peak height velocity (APHV) was inversely associated with adult IGF-I levels. IGF-I decreased by 3.7 ng/ml (95% confidence interval 1.0-6.4, P = 0.007) for each sd increase in APHV. Prepubertal childhood height and body mass index were both inversely associated with APHV (P trend < 0.001). APHV was positively associated with adult height and inversely associated with adult body mass index. Adjustment for childhood, adult anthropometry, and other lifestyle factors did not substantially alter the association between APHV and adult IGF-I. CONCLUSIONS: This is the first study to document an association between timing of puberty and adult IGF-I levels. A better understanding of life course determinants of the IGF system may provide new insights into disease etiology and primary prevention. PMID- 16720667 TI - Diagnosis of parathyroid tumors in familial isolated hyperparathyroidism with HRPT2 mutation: implications for cancer surveillance. AB - CONTEXT: Mutations of the HRPT2 gene have recently been implicated in the development of parathyroid carcinoma. OBJECTIVE: The objective of this study was early diagnosis of parathyroid tumor in a family with germline HRPT2 mutation. PATIENTS, METHODS, AND RESULTS: In a 40-yr-old male previously treated for parathyroid atypical adenoma, we screened the 17 translated HRPT2 exons and their exon-intron boundaries and found a germline frameshift mutation in exon 7 (685delAGAG) predicting a premature stop codon at nucleotides 767-769. Nine family members (age, 33.9 +/- 19.8 yr, mean +/- SD) also carry the mutation, but eight have had normal serum calcium. Biochemical and ultrasonographic evaluation uncovered a 27-yr-old hypercalcemic carrier niece with an atypical parathyroid adenoma, and a 43-yr-old normocalcemic carrier sister was found by ultrasonography to have an extrathyroidal nodule, which proved to be parathyroid carcinoma. The index case, 12 yr after surgery, was normocalcemic, but ultrasonography revealed an extrathyroidal nodule in the contralateral hemithyroid tissue that proved to be atypical adenoma. CONCLUSIONS: Our report confirms that germline mutations of HRPT2 gene may be associated with multiple parathyroid neoplasms. Our experience suggests that longitudinal surveillance by serum biochemistry alone may not be 100% sensitive, and addition of routine neck ultrasonography is a readily accepted adjunct that may facilitate earlier disease detection in some families. PMID- 16720668 TI - Testosterone use in men and its effects on bone health. A systematic review and meta-analysis of randomized placebo-controlled trials. AB - CONTEXT: Androgen-deficient men are at increased risk of osteoporosis. The extent to which testosterone can prevent and treat osteoporosis in men remains unclear. OBJECTIVE AND DESIGN: We performed a systematic review and meta-analysis of randomized placebo-controlled trials in men to estimate the effect of testosterone use on bone health outcomes. DATA SOURCES: The review encompassed librarian-designed search strategies using MEDLINE (1966 to March 2005), EMBASE (1988 to March 2005), and Cochrane CENTRAL (inception to March 2005); a review of reference lists from included studies; and content expert files. DATA COLLECTION: Independently and in duplicate, we assessed the methodological quality of the eligible trials and collected data on bone mineral density and bone fractures at the longest point of complete follow-up. DATA SYNTHESIS: We included eight trials enrolling 365 patients. Two trials followed patients for more than 1 yr. Meta analysis of these trials showed that, compared with placebo, im testosterone was associated with an 8% (95% confidence interval, 4%, 13%) gain in lumbar bone mineral density and transdermal testosterone had no significant impact. Testosterone use was associated with a nonsignificant 4% (95% confidence interval, -2%, 9%) gain in femoral neck bone mineral density with unexplained differences in results across trials (26% of these differences were not explained by chance alone). No trials measured or reported the effect of testosterone on fractures. CONCLUSIONS: Intramuscular testosterone moderately increased lumbar bone density in men; the results on femoral neck bone density are inconclusive. Without bone fracture data, the available trials offer weak and indirect inferences about the clinical efficacy of testosterone on osteoporosis prevention and treatment in men. PMID- 16720669 TI - Testosterone therapy in adult men with androgen deficiency syndromes: an endocrine society clinical practice guideline. AB - OBJECTIVE: The objective was to provide guidelines for the evaluation and treatment of androgen deficiency syndromes in adult men. PARTICIPANTS: The Task Force was composed of a chair, selected by the Clinical Guidelines Subcommittee of The Endocrine Society, five additional experts, a methodologist, and a professional writer. The Task Force received no corporate funding or remuneration. EVIDENCE: The Task Force used systematic reviews of available evidence to inform its key recommendations. The Task Force used consistent language and graphical descriptions of both the strength of recommendation and the quality of evidence, using the recommendations of the Grading of Recommendations, Assessment, Development, and Evaluation group. CONSENSUS PROCESS: Consensus was guided by systematic reviews of evidence and discussions during three group meetings, several conference calls, and e-mail communications. The drafts prepared by the panelists with the help of a professional writer were reviewed successively by The Endocrine Society's Clinical Guidelines Subcommittee, Clinical Affairs Committee, and Council. The version approved by the Council was placed on The Endocrine Society's web site for comments by members. At each stage of review, the Task Force received written comments and incorporated needed changes. CONCLUSIONS: We recommend making a diagnosis of androgen deficiency only in men with consistent symptoms and signs and unequivocally low serum testosterone levels. We suggest the measurement of morning total testosterone level by a reliable assay as the initial diagnostic test. We recommend confirmation of the diagnosis by repeating the measurement of morning total testosterone and in some patients by measurement of free or bioavailable testosterone level, using accurate assays. We recommend testosterone therapy for symptomatic men with androgen deficiency, who have low testosterone levels, to induce and maintain secondary sex characteristics and to improve their sexual function, sense of well-being, muscle mass and strength, and bone mineral density. We recommend against starting testosterone therapy in patients with breast or prostate cancer, a palpable prostate nodule or induration or prostate specific antigen greater than 3 ng/ml without further urological evaluation, erythrocytosis (hematocrit > 50%), hyperviscosity, untreated obstructive sleep apnea, severe lower urinary tract symptoms with International Prostate Symptom Score (IPSS) greater than 19, or class III or IV heart failure. When testosterone therapy is instituted, we suggest aiming at achieving testosterone levels during treatment in the mid-normal range with any of the approved formulations, chosen on the basis of the patient's preference, consideration of pharmacokinetics, treatment burden, and cost. Men receiving testosterone therapy should be monitored using a standardized plan. PMID- 16720670 TI - Impaired phagocytosis of capsular serotypes K1 or K2 Klebsiella pneumoniae in type 2 diabetes mellitus patients with poor glycemic control. AB - CONTEXT: Diabetes mellitus (DM) and capsular serotypes K1 and K2 Klebsiella pneumoniae have been identified as risk factors for liver abscess and complicated endophthalmitis. OBJECTIVE: The objective of this study was to determine whether poor glycemic control contributes to the development of capsular serotype K1 or K2 K. pneumoniae liver abscess. DESIGN AND SETTING: Neutrophil phagocytosis in patients with type 2 DM and nondiabetic controls was compared with isolates from liver abscess. Phagocytic rates of 18 K1/K2 and nine non-K1/K2 K. pneumoniae strains were evaluated by flow cytometry and electron microscopy. PATIENTS OR STUDY PARTICIPANTS: Forty patients with type 2 diabetes, 14 with good glycemic control, 26 with poor glycemic control, and 13 age-matched healthy normal subjects, were studied. MAIN OUTCOME MEASURES: Phagocytic rate of K. pneumoniae was measured. RESULTS: Phagocytosis of serotype K1/K2 isolates by neutrophils from diabetics was significantly less than normal controls (P < 0.01). Further analysis revealed that, in type 2 DM patients with poor glycemic control, phagocytosis of K1/K2 was remarkably impaired at 10 min (25.2 +/- 1.7 vs. 42.4 +/ 1.8%) and persisted until 60 min (51 +/- 1.2 vs. 59.4 +/- 1.4%; P < 0.01), but in type 2 DM patients with good glycemic control were similar at 10 min (38.2 +/- 1.7% vs. 42.4 +/- 1.8%) and at 60 min (57 +/- 0.3% vs. 59.4 +/- 1.4%; P = 0.2). No significant difference in the phagocytosis of non-K1/K2 K. pneumoniae among all subjects was observed. CONCLUSIONS: Poor glycemic control plays a role in impairing neutrophil phagocytosis of K1/K2 K. pneumoniae, but does not significantly affect the phagocytosis of non-K1/K2 K. pneumoniae. This study identifies poor glycemic control as a risk factor for susceptibility to serotype K1/K2 K. pneumoniae liver abscess and complicated endophthalmitis. PMID- 16720671 TI - Absorption of carbon dioxide during laparoscopy in children measured using a novel mass spectrometric technique. AB - BACKGROUND: Carbon dioxide (CO(2)) is absorbed during pneumoperitoneum and may cause adverse haemodynamic effects. The aim of this study was to measure the elimination of exogenous CO(2) during laparoscopy in children. METHODS: Ten children [27.6 (56.5) months; mean (SD)] undergoing laparoscopic and nine [24.5 (17.3) months] undergoing open surgery were studied. Breath samples were collected at the line for end-tidal CO(2) and analysed for (13)CO(2)/(12)CO(2) ratio expressed as deltaPDB (difference from standard), by isotope-ratio mass spectrometry. The proportion of absorbed CO(2) was calculated comparing exhaled (13)CO(2)/(12)CO(2) before and during CO(2) pneumoperitoneum. RESULTS: (13)CO(2)/(12)CO(2) in medical CO(2) was -32.7 (2.1) deltaPDB. (13)CO(2)/(12)CO(2) in breath of patients undergoing open procedures was -24.3 (2.4) deltaPDB at the start of operation and did not change during the operation (P > 0.2). (13)CO(2)/(12)CO(2) in breath of patients undergoing laparoscopy was 21.5 (5.4) deltaPDB at the start of insufflation, and decreased during pneumoperitoneum by 2.5 (1.6) deltaPDB, indicating absorption of exogenous CO(2). The percentage of expired CO(2) absorbed rose to 15.5 (7.7)% after 30 min of pneumoperitoneum and decreased rapidly after desufflation. CONCLUSION: After 10 min of laparoscopy 10-20% of expired CO(2) derives from the exogenous CO(2). CO(2) absorption can be measured using a simple mass spectrometric technique. PMID- 16720672 TI - Epidural catheter placement in children: comparing a novel approach using ultrasound guidance and a standard loss-of-resistance technique. AB - BACKGROUND: We report a prospective, randomized study to evaluate ultrasound guidance for epidural catheter placement in children 0-6 yr of age. METHODS: Epidural catheters were placed at lumbar or thoracic cord levels in 64 children undergoing major surgery, using either ultrasonography or loss-of-resistance (LOR) for guidance. Using a 5-10 MHz linear ultrasound probe, the neuraxial structures were identified, the skin-epidural depth and epidural space was measured, the advancing epidural catheter visualized, and the spread of local anaesthetic verifying catheter position was confirmed. Epidural placement procedures were analysed for bone contacts and speed of execution. Children under 6 months were analysed separately. RESULTS: Epidural placement involved bone contacts in 17% of children in the ultrasound group and 71% of children in the LOR group (P<0.0001). Epidurals were executed more swiftly in the ultrasound group [162 (75) s vs 234 (138) s; P<0.01]. Children under 6 months revealed a 0.9 correlation between skin-epidural depth and body weight. CONCLUSIONS: Ultrasonography is a useful aid to verify epidural placement of local anaesthetic agents and epidural catheters in children. Advantages include a reduction in bone contacts, faster epidural placement, direct visualization of neuraxial structures and the spread of local anaesthetic inside the epidural space. Ultrasound guidance requires additional training and good manual skills, and should only be used once experience in ultrasound-guided techniques of regional anaesthesia has been acquired. PMID- 16720673 TI - Skin conductance monitoring compared with bispectral index monitoring to assess emergence from general anaesthesia using sevoflurane and remifentanil. AB - BACKGROUND: Changes in skin conductance have previously been reported to correlate well with plasma levels of stress hormones and awakening stimuli. In this study, monitoring of skin conductance during emergence from general anaesthesia was compared with the monitoring of bispectral index (BIS). METHODS: Twenty-five patients undergoing minor elective surgery were investigated. The number of fluctuations in mean skin conductance (NFSC), BIS and haemodynamic parameters were recorded simultaneously. The performance of the monitoring devices to predict and distinguish between the clinical states 'steady-state anaesthesia', 'first reaction' and 'extubation' were compared using the method of prediction probability (P(K)) calculation. RESULTS: Both monitors showed similar performance in distinguishing between 'steady-state anaesthesia' vs 'first reaction' (P(K) NFSC 0.89; BIS 0.94) and 'steady-state anaesthesia' vs 'extubation' (P(K) NFSC 0.96; BIS 0.96). The response times of the monitors, to indicate the likelihood of 'first reaction', were not significantly different. CONCLUSIONS: NFSC, as a parameter of skin conductance, performed similarly to BIS in patients waking after a general anaesthetic. PMID- 16720674 TI - Acute respiratory distress syndrome with transiently impaired left ventricular function and Torsades de Pointes arrhythmia unmasking congenital long QT syndrome in a 25-yr-old woman. AB - We report a case of recurrent episodes of Torsades de Pointes arrhythmia in the setting of transiently impaired left ventricular ejection fraction, acute respiratory distress syndrome, transient hypokalaemia and QT-prolonging drugs, in a previously healthy 25-yr-old female patient. In the course of the clinical and genetic work-up this patient was newly diagnosed with a mutation in KCNH2 encoding the alpha-subunit of the human repolarizing potassium channel I(Kr). This case report illustrates the multivariate nature of long-QT syndrome, and emphasizes the usefulness of a pharmacological test for repolarization abnormalities. PMID- 16720675 TI - The effects of i.v. fentanyl administration on the minimum alveolar concentration of isoflurane in horses. AB - BACKGROUND: Fentanyl decreases the minimum alveolar concentration (MAC) of inhaled anaesthetics and has been used clinically to reduce the requirements of other anaesthetic drugs in humans and small animals. We hypothesized that i.v. fentanyl would decrease the MAC of isoflurane in horses in a dose-dependent manner. METHODS: Following determination of baseline MAC of isoflurane, fentanyl was administered i.v. to target plasma concentrations of 1, 8 and 16 ng ml(-1). Each horse was randomly assigned two of three target concentrations administered in ascending order. Loading and infusion doses for each horse were determined from previously derived individual pharmacokinetic values. Isoflurane MAC determination began 45 min after fentanyl administration at each target fentanyl concentration. Venous blood was collected at fixed intervals during the infusion for measurement of plasma fentanyl concentrations. RESULTS: Mean actual fentanyl plasma concentrations were 0 (baseline), and 0.72 (SD 0.26), 8.43 (3.22), and 13.31 (6.66) ng ml(-1) for the target concentrations of 1, 8 and 16 ng ml(-1), respectively. The corresponding isoflurane MAC values were a baseline of 1.57 (0.23), and 1.51 (0.24), 1.41 (0.23) and 1.37 (0.09)%, respectively. The fentanyl concentrations of 0.72 and 8.43 ng ml(-1) did not significantly alter the MAC of isoflurane, but an 18 (7)% ISO-MAC reduction was observed at the 13.31 ng ml(-1) concentration. CONCLUSIONS: These results cautiously encourage further study of fentanyl as an opioid anaesthetic adjunct to inhalant anaesthesia in horses. PMID- 16720676 TI - A comparison of the SNAP II and BIS XP indices during sevoflurane and nitrous oxide anaesthesia at 1 and 1.5 MAC and at awakening. AB - BACKGROUND: Monitoring level of consciousness during anaesthesia, with the ability to predict the intentional or unintentional return to consciousness, is desirable. The purpose of this study was to compare two processed electroencephalographic depth of anaesthesia monitors (SNAP II and BIS XP) during sevoflurane and sevoflurane/nitrous oxide anaesthesia. METHODS: In total, 42 subjects received an interscalene block, followed by general anaesthesia with sevoflurane or sevoflurane/nitrous oxide. The indices were recorded at baseline, at 1.5 and 1.0 minimum alveolar concentration (MAC) equivalents, and during emergence. RESULTS: The SNAP and BIS indices decreased from baseline at 1.5 and 1.0 MAC equivalents, but there was no difference within groups between subjects who received nitrous oxide and those who did not. The SNAP index returned to baseline by 1 min before awakening and was higher than baseline at eye opening, but the BIS index remained below baseline at awakening. There was a bias of -1 (95% CI: -3 to 1) between the SNAP and BIS at baseline; this increased to 21 (95% CI: 19-23) during maintenance of anaesthesia and was 6 (95% CI: 4-8) at awakening. CONCLUSIONS: The SNAP index tracks loss of consciousness and emergence from sevoflurane and sevoflurane/nitrous oxide anaesthesia. There is significant bias between the SNAP and BIS indices and therefore, the indices are not interchangeable. The SNAP index returns to baseline before awakening, whereas the BIS index remains below baseline at awakening, suggesting that the SNAP index may be more sensitive to unintentional awareness. PMID- 16720677 TI - N-ethyl-N-nitrosourea-based generation of mouse models for mutant G protein coupled receptors. AB - Chemical random mutagenesis techniques with the germ line supermutagen N-ethyl-N nitrosourea (ENU) have been established to provide comprehensive collections of mouse models, which were then mined and analyzed in phenotype-driven studies. Here, we applied ENU mutagenesis in a high-throughput fashion for a gene-driven identification of new mutations. Selected members of the large superfamily of G protein-coupled receptors (GPCR), melanocortin type 3 (Mc3r) and type 4 (Mc4r) receptors, and the orphan chemoattractant receptor GPR33, were used as model targets to prove the feasibility of this approach. Parallel archives of DNA and sperm from mice mutagenized with ENU were screened for mutations in these GPCR, and in vitro assays served as a preselection step before in vitro fertilization was performed to generate the appropriate mouse model. For example, mouse models for inherited obesity were established by selecting fully or partially inactivating mutations in Mc4r. Our technology described herein has the potential to provide mouse models for a GPCR dysfunction of choice within <4 mo and can be extended to other gene classes of interest. PMID- 16720678 TI - Sex-specific QTLs and interacting loci underlie salt-sensitive hypertension and target organ complications in Dahl S/jrHS hypertensive rats. AB - Sex-specific differences in polygenic (essential) hypertension are commonly attributed to the role of sex steroid hormone-receptor systems attenuating sex common disease mechanisms in premenopausal women. However, emerging observations indicate sex-specific genetic susceptibility in various traits, thus requiring systematic study. Here we report a comparative analysis of independent total genome scans for salt-sensitive hypertension susceptibility quantitative trait loci (QTLs) in male and female F2 [Dahl R/jrHS x S/jrHS] intercross rats exposed to high-salt (8% NaCl) rat diets. Hypertension was phenotyped with three quantitative traits: blood pressure (BP) elevation associated with increased hypertensive renal disease [glomerular injury score (GIS)] and increased cardiac mass [relative heart weight (RHW)] obtained 8-12 wk after high-salt challenge; 24 h nonstress, telemetric BP measurements were used. Although sex-common QTLs were detected for BP [chromosome (chr) 1-144.3 Mbp; chr 1-208.8 Mbp], GIS (chr 1-208.8 Mbp), and cardiac mass (chr 5-150.3 Mbp), most QTLs across the three phenotypes studied are gender specific as follows: female QTLs for BP (chr 2-106.7 Mbp, chr 2-181.7 Mbp, chr 5-113.9 Mbp, chr 5-146.7 Mbp, chr 12-12.8 Mbp), GIS (chr 15-59.6 Mbp), and RHW (chr 2-31.5 Mbp, chr 5-154.7 Mbp, chr 5-110.9 Mbp); male QTLs for BP (chr 2-196.7 Mbp, chr 11-48.0 Mbp, chr 20-35.7 Mbp), GIS (chr 6-3.3 Mbp, chr 20-40.7 Mbp), and RHW (chr 6-3.3 Mbp, chr 20-40.7 Mbp). Furthermore, interacting loci with significant linkage were detected only in female F2 intercross rats for BP and hypertensive renal disease. Comparative analyses revealed concordance of BP QTL peaks with previously reported rat model and human hypertension susceptibility genes and with BP QTLs in previous Dahl S-derived F2 intercross studies and also suggest strain-specific genetic modifiers of sex-specific determinants. Altogether, the data provide key experimental bases for sex specific investigation of mechanisms and intervention and prevention strategies for polygenic hypertension in humans. PMID- 16720679 TI - A step in the right direction. PMID- 16720681 TI - Stimulation of the caudal zona incerta is superior to stimulation of the subthalamic nucleus in improving contralateral parkinsonism. AB - Deep brain stimulation (DBS) has an increasing role in the treatment of idiopathic Parkinson's disease. Although, the subthalamic nucleus (STN) is the commonly chosen target, a number of groups have reported that the most effective contact lies dorsal/dorsomedial to the STN (region of the pallidofugal fibres and the rostral zona incerta) or at the junction between the dorsal border of the STN and the latter. We analysed our outcome data from Parkinson's disease patients treated with DBS between April 2002 and June 2004. During this period we moved our target from the STN to the region dorsomedial/medial to it and subsequently targeted the caudal part of the zona incerta nucleus (cZI). We present a comparison of the motor outcomes between these three groups of patients with optimal contacts within the STN (group 1), dorsomedial/medial to the STN (group 2) and in the cZI nucleus (group 3). Thirty-five patients with Parkinson's disease underwent MRI directed implantation of 64 DBS leads into the STN (17), dorsomedial/medial to STN (20) and cZI (27). The primary outcome measure was the contralateral Unified Parkinson's Disease Rating Scale (UPDRS) motor score (off medication/off stimulation versus off medication/on stimulation) measured at follow-up (median time 6 months). The secondary outcome measures were the UPDRS III subscores of tremor, bradykinesia and rigidity. Dyskinesia score, L-dopa medication reduction and stimulation parameters were also recorded. The mean adjusted contralateral UPDRS III score with cZI stimulation was 3.1 (76% reduction) compared to 4.9 (61% reduction) in group 2 and 5.7 (55% reduction) in the STN (P-value for trend <0.001). There was a 93% improvement in tremor with cZI stimulation versus 86% in group 2 versus 61% in group 1 (P-value = 0.01). Adjusted 'off-on' rigidity scores were 1.0 for the cZI group (76% reduction), 2.0 for group 2 (52% reduction) and 2.1 for group 1 (50% reduction) (P-value for trend = 0.002). Bradykinesia was more markedly improved in the cZI group (65%) compared to group 2 (56%) or STN group (59%) (P-value for trend = 0.17). There were no statistically significant differences in the dyskinesia scores, L-dopa medication reduction and stimulation parameters between the three groups. Stimulation related complications were seen in some group 2 patients. High frequency stimulation of the cZI results in greater improvement in contralateral motor scores in Parkinson's disease patients than stimulation of the STN. We discuss the implications of this finding and the potential role played by the ZI in Parkinson's disease. PMID- 16720680 TI - Gene expression and benefit of chemotherapy in women with node-negative, estrogen receptor-positive breast cancer. AB - PURPOSE: The 21-gene recurrence score (RS) assay quantifies the likelihood of distant recurrence in women with estrogen receptor-positive, lymph node-negative breast cancer treated with adjuvant tamoxifen. The relationship between the RS and chemotherapy benefit is not known. METHODS: The RS was measured in tumors from the tamoxifen-treated and tamoxifen plus chemotherapy-treated patients in the National Surgical Adjuvant Breast and Bowel Project (NSABP) B20 trial. Cox proportional hazards models were utilized to test for interaction between chemotherapy treatment and the RS. RESULTS: A total of 651 patients were assessable (227 randomly assigned to tamoxifen and 424 randomly assigned to tamoxifen plus chemotherapy). The test for interaction between chemotherapy treatment and RS was statistically significant (P = .038). Patients with high-RS (> or = 31) tumors (ie, high risk of recurrence) had a large benefit from chemotherapy (relative risk, 0.26; 95% CI, 0.13 to 0.53; absolute decrease in 10 year distant recurrence rate: mean, 27.6%; SE, 8.0%). Patients with low-RS (< 18) tumors derived minimal, if any, benefit from chemotherapy treatment (relative risk, 1.31; 95% CI, 0.46 to 3.78; absolute decrease in distant recurrence rate at 10 years: mean, -1.1%; SE, 2.2%). Patients with intermediate-RS tumors did not appear to have a large benefit, but the uncertainty in the estimate can not exclude a clinically important benefit. CONCLUSION: The RS assay not only quantifies the likelihood of breast cancer recurrence in women with node negative, estrogen receptor-positive breast cancer, but also predicts the magnitude of chemotherapy benefit. PMID- 16720683 TI - Effect of cytochromes P450 chemical inhibitors and monoclonal antibodies on human liver microsomal esterase activity. AB - Selective and nonselective cytochromes P450 (P450) chemical inhibitors and monoclonal antibodies (mAbs) are routinely used to determine the contribution of P450 enzymes involved in the biotransformation of a drug. A fluorometric assay has been established using fluorescein diacetate as a model substrate to determine the effect of some commonly used P450 inhibitors and mAbs on human liver microsomal esterase activity. Of those inhibitors studied, only alpha naphthoflavone, clotrimazole, ketoconazole, miconazole, nicardipine, and verapamil significantly inhibited human liver microsomal esterase activity, with apparent IC50 values of 18.0, 20.5, 6.5, 15.0, 19.4, and 5.4 microM, respectively. All of these showed > or =20% inhibition of human liver microsomal esterase activity at concentrations typically used for P450 reaction phenotyping studies, with clotrimazole, miconazole, nicardipine, and verapamil showing >60% inhibition. Unlike the chemical inhibitors, no inhibition of human liver microsomal esterase activity was observed in the presence of mAb to CYP1A2, 2C8, 2C9, 2C19, 2D6, and 3A4. These results suggest that P450 chemical inhibitors are capable of inhibiting human liver microsomal esterase activity and should not be used to assess the role of P450 enzymes in the biotransformation of esters. The lack of inhibition of human liver microsomal esterase activity by P450-specific monoclonal antibodies suggests that they may be used to assess the role of P450 enzymes in the biotransformation of esters. Additional experiments to assess the contribution of oxidative enzymes in the metabolism of esters may include incubations in the presence and absence of beta-nicotinamide adenine dinucleotide 2'-phosphate reduced. PMID- 16720682 TI - Clinical findings and diagnostic tests in the MV2 subtype of sporadic CJD. AB - Atypical clinical course and low sensitivity of established diagnostic tests are the main diagnostic problems in the MV2 subtype of sporadic Creutzfeldt-Jakob disease (sCJD). Clinical symptoms and signs, MRI, EEG and biochemical CSF markers were studied in 26 patients. Histological findings were semiquantitatively evaluated. Compared with typical sCJD, the disease duration was prolonged (median 12 months). Dementia, ataxia and psychiatric symptoms were present in all patients. Extrapyramidal signs were observed in 88%. T2-weighted MRI showed basal ganglia hyperintensities in 90%. Increased thalamic signal intensity was detected in 88% on diffusion-weighted MRI. Increased CSF tau-protein was found in 83%, and the 14-3-3 test was positive in 76%. The EEG revealed periodic sharp wave complexes in only two patients. Kuru plaques, severe thalamic and basal ganglia gliosis and spongiform changes, and neuronal loss in the pulvinar were the prominent histological features. At least one of the three diagnostic tests (MRI, tau- and 14-3-3 protein) supported the clinical diagnosis in all patients. MRI was the most sensitive of the diagnostic tests applied. Thalamic hyperintensities were observed unusually frequently. Prolonged disease duration, early and prominent psychiatric symptoms, absence of typical EEG, thalamic hyperintensities on MRI and relatively low 14-3-3 protein sensitivity may be suspicious for variant CJD. However, distinct sensory symptoms and young age at onset, which are often found in the latter, are not common in the MV2 subtype, and the pulvinar sign was observed in only one case. PMID- 16720684 TI - N-glucuronidation of perfluorooctanesulfonamide by human, rat, dog, and monkey liver microsomes and by expressed rat and human UDP-glucuronosyltransferases. AB - N-Alkylperfluorooctanesulfonamides have been used in a range of industrial and commercial applications. Perfluorooctanesulfonamide (FOSA) is a major metabolite of N-alkylperfluorooctanesulfonamides and has a long half-life in animals and in the environment and is biotransformed to FOSA N-glucuronide. The objective of this study was to identify and characterize the human and experimental animal liver UDP-glucuronosyltransferases (UGTs) that catalyze the N-glucuronidation of FOSA. The results showed that pooled human liver and rat liver microsomes had high N-glucuronidation activities. Expressed rat UGT1.1, UGT2B1, and UGT2B12 in HK293 cells catalyzed the N-glucuronidation of FOSA but at rates that were lower than those observed in rat liver microsomes. Of the 10 expressed human UGTs (1A1, 1A3, 1A4, 1A6, 1A9, 2B4, 2B7, 2B15, and 2B17) studied, only hUGT2B4 and hUGT2B7 catalyzed the N-glucuronidation of FOSA. The kinetics of N-glucuronidation of FOSA by rat liver microsomes and by hUGT2B4/7 was consistent with a single-enzyme Michaelis-Menten model, whereas human liver microsomes showed sigmoidal kinetics. These data show that rat liver UGT1.1, UGT2B1, and UGT2B12 catalyze the N glucuronidation of FOSA, albeit at low rates, and that hUGT2B4 and hUGT2B7 catalyze the N-glucuronidation of FOSA. PMID- 16720685 TI - Cardiac magnetic resonance perfusion imaging for the functional assessment of coronary artery disease: a comparison with coronary angiography and fractional flow reserve. AB - AIMS: Cardiac magnetic resonance perfusion imaging (CMRI) is a promising technique for non-invasive measurement of myocardial perfusion reserve. Fractional flow reserve (FFR) is an established invasive method for functional assessment of coronary artery disease (CAD). To prospectively assess the diagnostic value of CMRI for the detection of haemodynamically significant coronary lesions, compared with coronary angiography (CA) and FFR. METHODS AND RESULTS: Forty-three patients with suspected or known CAD underwent CA, CMRI, and FFR measurement. First pass magnetic resonance perfusion examination was performed during hyperaemia (140 microg/kg/min adenosine over 6 min) and at rest. One hundred and twenty-nine perfusion territories were assessed by semi quantitative evaluation of signal intensity-time curves using the myocardial perfusion reserve index (MPRI) [upslope(stress(corrected))/upslope(rest(corrected))]. Perfusion territories were categorized as normal (coronary stenosis < or = 50%), intermediate (stenosis > 50% and FFR > 0.75), or severe (stenosis > 50% and FFR < or = 0.75 or total occlusion). MPRI values (+/-SD) were significantly different between the three categories [normal, 2.2 +/- 0.5 vs. intermediate, 1.8 +/- 0.5 (P = 0.005) and intermediate vs. severe, 1.2 +/- 0.3 (P < 0.001)]. An MPRI cut-off value of 1.5 (derived from receiver operating characteristics analysis) distinguished haemodynamically relevant (severe) from non-relevant (normal and intermediate) stenoses with a sensitivity of 88% (CI 74-100%) and a specificity of 90% (CI 84 96%). CONCLUSION: In contrast to earlier studies that compared CMRI with morphological examination (CA) alone, the present study compared CMRI with CA plus a standard invasive functional assessment (FFR) and demonstrated that CMRI is able to distinguish haemodynamically relevant from non-relevant coronary lesions with a high sensitivity and specificity and may therefore contribute to clinical decision-making. PMID- 16720686 TI - Apical ballooning syndrome or takotsubo cardiomyopathy: a systematic review. AB - AIMS: To clarify the major features of the apical ballooning syndrome, we performed a systematic review of the existing literature. METHODS AND RESULTS: Review of all relevant case series using the MEDLINE and EMBASE databases resulted in the identification of 14 studies. These studies suggest that the apical ballooning syndrome accounts for approximately 2.0% of ST-segment elevation infarcts, with most cases described in post-menopausal women. The most common clinical presentations are chest pain and dyspnoea, reported in 67.8 and 17.8% of the patients, respectively. Cardiogenic shock (4.2% of the patients) and ventricular fibrillation (1.5%) were not infrequent. ST-segment elevation was reported in 81.6% of the patients, T wave abnormalities in 64.3%, and Q waves in 31.8%. Cardiac biomarkers were usually mildly elevated, as reported in 86.2% of the patients. Typically, patients had left ventricular (LV) dysfunction on admission, with mean ejection fraction ranging from 20 to 49%. However, over a period of days to weeks, all patients experienced dramatic improvement in LV function. The onset of symptoms was often preceded by emotional (26.8%) or physical stress (37.8%). Norepinephrine concentration was elevated in 74.3% of the patients. Prognosis was generally excellent, with full recovery in most patients. In-hospital mortality was 1.1%. Only 3.5% of the patients experienced a recurrence. CONCLUSION: Clinicians should consider this syndrome in the differential diagnosis of patients presenting with chest pain, especially in post menopausal women with a recent history of emotional or physical stress. PMID- 16720687 TI - Comparison of real-time tri-plane and conventional 2D dobutamine stress echocardiography for the assessment of coronary artery disease. AB - AIMS: Although dobutamine stress echocardiography (DSE) is an accepted tool for the diagnosis of coronary artery disease (CAD), it requires subsequent image acquisitions of the left ventricle (LV) in order to visualize all segments. This makes the procedure relatively time-consuming and might limit its accuracy. With the introduction of matrix array transducers, the real-time simultaneous acquisition of all LV segments has become possible using multi-plane imaging. The purpose of this study was: (i) to test the feasibility and efficiency of real time tri-plane (RT3P) imaging during DSE, (ii) to compare the accuracy of RT3P DSE in detecting CAD using coronary angiography as the reference method. METHODS AND RESULTS: Thirty-six patients suspected of CAD were prospectively enrolled. Both conventional two-dimensional (2D) and RT3P imaging were performed during a DSE protocol. Coronary angiography was performed within 24 h. Ultrasound data were acquired at each stage of the DSE. The total effective acquisition time for RT3P imaging was significantly shorter (55+/-29 vs. 137+/-63 s, P<0.001). Data yield was similar for both methods (2D: 98% vs. 3D: 97%). Overall sensitivity (93%), specificity (75%), and accuracy (89%) were identical between both methods. On a segmental level, the sensitivity, specificity, and accuracy of the RT3P and the 2D DSE were similar. CONCLUSION: RT3P imaging fastens the DSE protocol without compromising the accuracy for the diagnosis of CAD. This could facilitate a more wide-spread use of DSE and therefore contributes positively to its routine clinical acceptance. PMID- 16720688 TI - Women, 'non-specific' chest pain, and normal or near-normal coronary angiograms are not synonymous with favourable outcome. PMID- 16720689 TI - Regression of left ventricular mass and wall thickness after cardiac resynchronization therapy: proof of pathophysiological concept. PMID- 16720690 TI - The demystification of magnetic resonance imaging? PMID- 16720691 TI - Persistent chest pain predicts cardiovascular events in women without obstructive coronary artery disease: results from the NIH-NHLBI-sponsored Women's Ischaemia Syndrome Evaluation (WISE) study. AB - AIMS: Women with chest pain but without obstructive coronary artery disease (CAD) are considered at low risk for cardiovascular (CV) events, but half continue to experience debilitating chest pain over many years. This study compared CV outcomes in women with persistent chest pain (PChP) vs. those without PChP. METHODS AND RESULTS: We studied 673 Women's Ischaemia Syndrome Evaluation (WISE) participants with chest pain undergoing coronary angiography for suspected myocardial ischaemia and at least 1 year of follow-up. PChP was defined as self reported continuing chest pain after 1 year. Events occurring after that year were recorded for a median of 5.2 years. We compared CV event rates for women with and without PChP in subgroups with and without obstructive CAD. The median age was 58 years, 20% were racial minorities, 45% had PChP, 39% had obstructive CAD. Among women without CAD, those with PChP had more than twice the rate of composite CV events (P = 0.03), that included non-fatal myocardial infarctions (P = 0.11), strokes (P = 0.03), congestive heart failure (P = 0.38), and CV deaths (P = 0.73), compared with those without PChP. In women with CAD, there was no difference in composite CV events in those with and without PChP (P = 0.72). CONCLUSION: Among women undergoing coronary angiography for suspected myocardial ischaemia, PChP in women with no obstructive CAD predicted adverse CV outcomes. Such women might benefit from additional evaluation and aggressive risk factor modification therapy. PMID- 16720692 TI - The correlation between protein stability and dipole moment: a critical test. AB - Improving the stability of proteins is a major aim in basic and applied protein science. Querol and coworkers calculated changes in the quasi-electric dipole moment of a protein and used it as a simple criterion to predict stabilizing charge mutations. They employed this method to propose for the bacterial cold shock protein Bc-Csp a number of charge mutations that should have a strong influence on stability. We produced eight variants of Bc-Csp with such mutations and measured their stabilities experimentally. However, we could not find a correlation between the stability and the quasi dipole moment of these variants. Possibly, the quasi dipole moment reflects only a secondary aspect of the changes that are caused by charge mutations in a protein. PMID- 16720693 TI - A system for concomitant overexpression of four periplasmic folding catalysts to improve secretory protein production in Escherichia coli. AB - Although Escherichia coli is in wide use for preparative protein expression, problems with the folding of the recombinant gene product and protein aggregation are frequently encountered. Apart from cytoplasmic expression, this is also true for secretion into the bacterial periplasm, the method of choice for the production of proteins that carry structural disulfide bonds. Here we report the construction of the helper plasmid pTUM4, which effects overexpression of four established periplasmic chaperones and folding catalysts: the thiol-disulfide oxidoreductases DsbA and DsbC that catalyze the formation and isomerization of disulfide bridges and the peptidyl-prolyl cis/trans-isomerases with chaperone activity, FkpA and SurA. pTUM4 carries a p15a origin of replication and a chloramphenicol resistance gene and, thus, it is compatible with many conventional expression vectors that use the ColEI origin and an ampicillin resistance. Its positive effects on the yield of soluble recombinant protein and the homogeneity of disulfide pattern are illustrated here using the human plasma retinol-binding protein as well as the extracellular carbohydrate recognition domain of the dendritic cell membrane receptor DC-SIGN. Hence, pTUM4 represents a novel helper vector which complements existing cytosolic chaperone coexpression plasmids and should be useful for the functional secretion of various recombinant proteins with hampered folding efficiency. PMID- 16720694 TI - High rate of recent intron gain and loss in simultaneously duplicated Arabidopsis genes. AB - We examined the gene structure of a set of 2563 Arabidopsis thaliana paralogous pairs that were duplicated simultaneously 20-60 MYA by tetraploidy. Out of a total of 23,164 introns in these genes, we found that 10,004 pairs have been conserved and 578 introns have been inserted or deleted in the time since the duplication event. This intron insertion/deletion rate of 2.7 x 10(-3) to 9.1 x 10(-4) per site per million years is high in comparison to previous studies. At least 56 introns were gained and 39 lost based on parsimony analysis of the phylogenetic distribution of these introns. We found weak evidence that genes undergoing intron gain and loss are biased with respect to gene ontology terms. Gene pairs that experienced at least 2 intron insertions or deletions show evidence of enrichment for membrane location and transport and transporter activity function. We do not find any relationship of intron flux to expression level or G + C content of the gene. Detection of a bias in the location of intron gains and losses within a gene depends on the method of measurement: an intragene method indicates that events (specifically intron losses) are biased toward the 3' end of the gene. Despite the relatively recent acquisition of these introns, we found only one case where we could identify the mechanism of intron origin- the TOUCH3 gene has experienced 2 tandem, partial, internal gene duplications that duplicated a preexisting intron and also created a novel, alternatively spliced intron that makes use of a duplicated pair of cryptic splice sites. PMID- 16720695 TI - The evolutionary origin of an altruistic gene. AB - Although the conditions favoring altruism are being increasingly understood, the evolutionary origins of the genetic basis for this behavior remain elusive. Here, we show that reproductive altruism (i.e., a sterile soma) in the multicellular green alga, Volvox carteri, evolved via the co-option of a life-history gene whose expression in the unicellular ancestor was conditioned on an environmental cue (as an adaptive strategy to enhance survival at an immediate cost to reproduction) through shifting its expression from a temporal (environmentally induced) into a spatial (developmental) context. The gene belongs to a diverged and structurally heterogeneous multigene family sharing a SAND-like domain (a DNA binding module involved in gene transcription regulation). To our knowledge, this is the first example of a social gene specifically associated with reproductive altruism, whose origin can be traced back to a solitary ancestor. These findings complement recent proposals that the differentiation of sterile castes in social insects involved the co-option of regulatory networks that control sequential shifts between phases in the life cycle of solitary insects. PMID- 16720696 TI - Dependence among sites in RNA evolution. AB - Although probabilistic models of genotype (e.g., DNA sequence) evolution have been greatly elaborated, less attention has been paid to the effect of phenotype on the evolution of the genotype. Here we propose an evolutionary model and a Bayesian inference procedure that are aimed at filling this gap. In the model, RNA secondary structure links genotype and phenotype by treating the approximate free energy of a sequence folded into a secondary structure as a surrogate for fitness. The underlying idea is that a nucleotide substitution resulting in a more stable secondary structure should have a higher rate than a substitution that yields a less stable secondary structure. This free energy approach incorporates evolutionary dependencies among sequence positions beyond those that are reflected simply by jointly modeling change at paired positions in an RNA helix. Although there is not a formal requirement with this approach that secondary structure be known and nearly invariant over evolutionary time, computational considerations make these assumptions attractive and they have been adopted in a software program that permits statistical analysis of multiple homologous sequences that are related via a known phylogenetic tree topology. Analyses of 5S ribosomal RNA sequences are presented to illustrate and quantify the strong impact that RNA secondary structure has on substitution rates. Analyses on simulated sequences show that the new inference procedure has reasonable statistical properties. Potential applications of this procedure, including improved ancestral sequence inference and location of functionally interesting sites, are discussed. PMID- 16720697 TI - Evolution of the cichlid visual palette through ontogenetic subfunctionalization of the opsin gene arrays. AB - The evolution of cone opsin genes is characterized by a dynamic process of gene birth and death through gene duplication and loss. However, the forces governing the retention and death of opsin genes are poorly understood. African cichlid fishes have a range of ecologies, differing in habitat and foraging style, which make them ideal for examining the selective forces acting on the opsin gene family. In this work, we present data on the riverine cichlid, Oreochromis niloticus, which is an ancestral outgroup to the cichlid adaptive radiations in the Great African lakes. We identify 7 cone opsin genes with several instances of gene duplication. We also characterize the spectral sensitivities of these genes through reconstitution of visual pigments. Peak absorbances demonstrate that each tilapia cone opsin gene codes for a spectrally distinct visual pigment: SWS1 (360 nm), SWS2b (423 nm), SWS2a (456 nm), Rh2b (472 nm), Rh2a beta (518 nm), Rh2a alpha (528 nm), and LWS (561 nm). Furthermore, quantitative reverse transcription polymerase chain reaction at 3 ontogenetic time points demonstrates that although only 4 genes (SWS2a, Rh2a alpha and beta, and LWS) are expressed in adults, mRNAs for the other genes are all expressed during ontogeny. Therefore, subfunctionalization through differential ontogenetic expression may be a key mechanism for preservation of opsin genes. The distinct peak absorbances of these preserved opsin genes provide a palette from which selection creates the diverse visual sensitivities found among the cichlid species of the lacustrine adaptive radiations. PMID- 16720698 TI - Soft-lithographic replication of 3D microstructures with closed loops. AB - There is growing interest in lithographic technologies for creating 3D microstructures. Such techniques are generally serial in nature, prohibiting the mass production of devices. Soft-lithographic techniques show great promise for simple and rapid replication of arrays of microstructures but have heretofore not been capable of direct replication of structures with closed loops. We demonstrate that 3D microstructures created with multiphoton absorption polymerization can be replicated by using microtransfer molding to afford complex daughter structures containing closed loops. This method relieves many of the topological constraints of soft lithography, paving the way for the large-scale replication of true 3D microstructures. PMID- 16720699 TI - The dsRNA binding site of human Toll-like receptor 3. AB - Pathogen recognition by Toll-like receptors (TLRs) initiates innate immune responses that are essential for inhibiting pathogen dissemination and for the development of acquired immunity. The TLRs recognize pathogens with their N terminal ectodomains (ECD), but the molecular basis for this recognition is not known. Recently we reported the x-ray structure for unliganded TLR3-ECD; however, it has proven difficult to obtain a crystal structure of TLR3 with its ligand, dsRNA. We have now located the TLR3 ligand binding site by mutational analysis. More than 50 single-residue mutations have been generated throughout the TLR3 ECD, but only two, H539E and N541A, resulted in the loss of TLR3 activation and ligand binding functions. These mutations locate the dsRNA binding site on the glycan-free, lateral surface of TLR3 toward the C terminus and suggest a model for dsRNA binding and TLR3 activation. PMID- 16720700 TI - Plant-derived anti-Lewis Y mAb exhibits biological activities for efficient immunotherapy against human cancer cells. AB - Although current demands for therapeutic mAbs are growing quickly, production methods to date, including in vitro mammalian tissue culture and transgenic animals, provide only limited quantities at high cost. Several tumor-associated antigens in tumor cells have been identified as targets for therapeutic mAbs. Here we describe the production of mAb BR55-2 (IgG2a) in transgenic plants that recognizes the nonprotein tumor-associated antigen Lewis Y oligosaccharide overexpressed in human carcinomas, particularly breast and colorectal cancers. Heavy and light chains of mAb BR55-2 were expressed separately and assembled in plant cells of low-alkaloid tobacco transgenic plants (Nicotiana tabacum cv. LAMD609). Expression levels of plant-derived mAb (mAbP) were high (30 mg/kg of fresh leaves) in T1 generation plants. Like the mammalian-derived mAbM, the plant mAbP bound specifically to both SK-BR3 breast cancer cells and SW948 colorectal cancer cells. The Fc domain of both mAbP and mAbM showed the similar binding to FcgammaRI receptor (CD64). Comparable levels of cytotoxicity against SK-BR3 cells were also shown for both mAbs in antibody-dependent cell-mediated cytotoxicity assay. Furthermore, plant-derived BR55-2 efficiently inhibited SW948 tumor growth xenografted in nude mice. Altogether, these findings suggest that mAbP originating from low-alkaloid tobacco exhibit biological activities suitable for efficient immunotherapy. PMID- 16720701 TI - Fragments of ATP synthase mediate plant perception of insect attack. AB - Plants can perceive a wide range of biotic attackers and respond with targeted induced defenses. Specificity in plant non-self-recognition occurs either directly by perception of pest-derived elicitors or indirectly through resistance protein recognition of host targets that are inappropriately proteolyzed. Indirect plant perception can occur during interactions with pathogens, yet evidence for analogous events mediating the detection of insect herbivores remains elusive. Here we report indirect perception of herbivory in cowpea (Vigna unguiculata) plants attacked by fall armyworm (Spodoptera frugiperda) larvae. We isolated and identified a disulfide-bridged peptide (+ICDINGVCVDA-), termed inceptin, from S. frugiperda larval oral secretions that promotes cowpea ethylene production at 1 fmol leaf(-1) and triggers increases in the defense-related phytohormones salicylic acid and jasmonic acid. Inceptins are proteolytic fragments of chloroplastic ATP synthase gamma-subunit regulatory regions that mediate plant perception of herbivory through the induction of volatile, phenylpropanoid, and protease inhibitor defenses. Only S. frugiperda larvae that previously ingested chloroplastic ATP synthase gamma-subunit proteins and produced inceptins significantly induced cowpea defenses after herbivory. Digestive fragments of an ancient and essential plant enzyme, inceptin functions as a potent indirect signal initiating specific plant responses to insect attack. PMID- 16720702 TI - Zinc binding to a regulatory zinc-sensing domain monitored in vivo by using FRET. AB - We have generated probes of metal binding to zinc fingers (ZFs) that provide tools to study zinc trafficking in vivo. In this study, we used these probes to examine zinc binding by the Zap1 transcription factor of Saccharomyces cerevisiae. Zap1 contains two zinc-regulated activation domains (ADs), AD1 and AD2. AD2 is located within two C2H2 ZFs, ZF1 and ZF2. Studies have indicated that apoAD2 activates transcription and zinc binding to ZF1 and that ZF2 forms an interacting-finger-pair structure that is necessary to inhibit AD function. A related structural finger pair, ZF3 and ZF4, is found in the Zap1 DNA binding domain. In vitro studies indicated that, although the ZF1/2 and ZF3/4 finger pairs bind zinc with similar affinities, zinc that was bound to ZF1/2 was much more labile. We examined the properties of Zap1 ZFs in vivo by FRET. ZF pairs were flanked by enhanced yellow fluorescent protein and enhanced cyan fluorescent protein, allowing detection of zinc-induced conformation changes by FRET. By using these reporters, we found that ZF1/2 and ZF3/4 showed similar responses to zinc under steady-state conditions in vivo. In contrast, ZF1/2 zinc binding was significantly more labile than was ZF3/4. Also, ZF1/2 accumulated in an apo form that could rapidly bind zinc, whereas the ZF3/4 pair did not. Last, we show that these properties are evolutionarily conserved indicating their importance to Zap1 function. These results indicate that the kinetic lability of ZF1/2 in vivo is a key component of Zap1 zinc responsiveness. PMID- 16720703 TI - Carbamazepine toxicity induced by lopinavir/ritonavir and nelfinavir. AB - OBJECTIVE: To present a case of carbamazepine toxicity induced by lopinavir/ritonavir and nelfinavir. CASE SUMMARY: A 50-year-old HIV-positive male developed excessive drowsiness secondary to carbamazepine when an antiretroviral regimen containing lopinavir/ritonavir was introduced. The carbamazepine serum concentration increased 46%. Subsequently, the patient developed a possible adverse skin reaction to his antiretrovirals and was hospitalized. The protease inhibitor was changed to nelfinavir. Within 3 days, the patient again developed excessive drowsiness and became unsteady on his feet. This time, the carbamazepine serum concentration had increased by 53%. In both instances, the carbamazepine dosage was decreased by 33%, which resulted in resolution of symptoms. DISCUSSION: Carbamazepine undergoes extensive hepatic metabolism. The major metabolic pathway involves oxidation of carbamazepine via CYP3A4 to an active metabolite, carbamazepine-10,11-epoxide. Protease inhibitors are well known CYP3A4 inhibitors. Other cases of carbamazepine toxicity secondary to protease inhibitors are reviewed. A MEDLINE search (1966-May 2006) revealed 4 cases of carbamazepine toxicity secondary to antiretrovirals. Carbamazepine serum concentrations increased two- to threefold from baseline. Vertigo, drowsiness, disorientation, ataxia, and vomiting occurred within 12 hours to 2 months, which resolved with reduction of the carbamazepine dosage. CONCLUSIONS: An objective causality assessment suggests that our patient became drowsy and unsteady on his feet secondary to a carbamazepine-protease inhibitor interaction. Lopinavir/ritonavir and nelfinavir may decrease carbamazepine metabolism, causing an elevation in carbamazepine serum concentrations. Carbamazepine toxicity may be prevented by reducing the carbamazepine dosage by 25-50% when protease inhibitors are introduced. A carbamazepine serum concentration should be repeated 3-5 days after the protease inhibitors are started. PMID- 16720704 TI - Development of a teaching certificate program utilizing distance education. PMID- 16720705 TI - Impact of linezolid on economic outcomes and determinants of cost in a clinical trial evaluating patients with MRSA complicated skin and soft-tissue infections. AB - BACKGROUND: In clinical trials, linezolid has demonstrated higher clinical cure rates and shorter hospital duration for patients than has vancomycin for the treatment of complicated skin and soft-tissue infections (cSSTIs). OBJECTIVE: To assess economic outcomes of linezolid versus vancomycin and evaluate determinants of treatment costs for cSSTIs. METHODS: Economic data were obtained from US subjects enrolled in a multinational, open-label, clinical trial of cSSTIs caused by suspected or proven methicillin-resistant Staphylococcus aureus (MRSA). Subjects were randomized to receive intravenous or oral linezolid or intravenous vancomycin for 7-21 days. Costs for each patient were evaluated by applying nationally representative per diem hospital costs by hospital ward. Intravenous administration costs were applied to the duration of intravenous treatment. Factors contributing to the cost of therapy were evaluated using multivariate regression analysis. RESULTS: Seven hundred seventeen US patients were included in the study. Demographics were similar between treatment groups. Length of stay and duration of intravenous therapy were shorter for linezolid-treated patients. Mean +/- SD cost for intent-to-treat population patients treated with linezolid versus vancomycin was 4865 US dollars +/- 4367 versus 5738 US dollars +/- 5190, respectively (p = 0.017), and in the MRSA population was 4881 US dollars +/- 3987 versus 6006 US dollars +/- 5039, respectively (p = 0.041). Factors significantly associated with increased cost included vancomycin therapy, age, and comorbidities, including diabetes. After adjusting for all other factors, treatment with linezolid was associated with significantly lower treatment costs compared with vancomycin. CONCLUSIONS: Linezolid therapy was associated with improved clinical outcomes and significantly lower treatment costs than was vancomycin. The largest cost advantage was demonstrated in patients with documented MRSA cSSTIs. PMID- 16720706 TI - Physical and chemical stability of pemetrexed in infusion solutions. AB - BACKGROUND: Pemetrexed is a multitargeted, antifolate, antineoplastic agent that is indicated for single-agent use in locally advanced or metastatic non-small cell lung cancer after prior chemotherapy and in combination with cisplatin for the treatment of malignant pleural mesothelioma not treatable by surgery. Currently, there is no information on the long-term stability of pemetrexed beyond 24 hours. OBJECTIVE: To evaluate the longer-term physical and chemical stability of pemetrexed 2, 10, and 20 mg/mL in polyvinyl chloride (PVC) bags of dextrose 5% injection and NaCl 0.9% injection. METHODS: Triplicate samples of pemetrexed were prepared in the concentrations and infusion solutions required. Evaluations for physical and chemical stability were performed initially and over 2 days at 23 degrees C protected from light and exposed to fluorescent light, and over 31 days of storage at 4 degrees C protected from light. Physical stability was assessed using turbidimetric and particulate measurement as well as visual observation. Chemical stability was evaluated by HPLC. RESULTS: All pemetrexed solutions remained chemically stable, with little or no loss of pemetrexed over 2 days at 23 degrees C, protected from light and exposed to fluorescent light, and over 31 days of storage at 4 degrees C, protected from light. The room temperature samples were physically stable throughout the 48 hour test period. However, pemetrexed admixtures developed large numbers of microparticulates during refrigerated storage exceeding 24 hours. CONCLUSIONS: Pemetrexed is chemically stable for 2 days at room temperature and 31 days refrigerated in dextrose 5% injection and NaCl 0.9% injection. However, substantial numbers of microparticulates may form in pemetrexed diluted in the infusion solutions in PVC bags, especially during longer periods of refrigerated storage. Limiting the refrigerated storage period to the manufacturer-recommended 24 hours will limit particulate formation. PMID- 16720707 TI - Withdrawal from lorazepam in critically ill children. AB - BACKGROUND: Sedatives are used in critically ill children to facilitate mechanical ventilation. Although tolerance and withdrawal are associated with use of sedatives, information about withdrawal from benzodiazepines in children is limited. OBJECTIVE: To document the occurrence of lorazepam withdrawal in critically ill children and identify predictors for the development of withdrawal. METHODS: This prospective, investigational, open-label study enrolled pediatric patients receiving a continuous infusion of lorazepam for at least 72 hours. The lorazepam dosage was tapered in a uniform fashion over 6 days by decreasing the total daily dose by 50% every other day on 3 occasions; it was then discontinued. The occurrence of withdrawal from lorazepam was determined by pediatric intensive care unit attending physicians based on clinical judgment. Patients were assessed for withdrawal twice daily beginning 48 hours after the initiation of the lorazepam taper. Assessments were continued for 72 hours after lorazepam discontinuation or until the patient experienced withdrawal, whichever came first. Patient demographic, sedative dosing, and lorazepam serum concentration data were collected to identify risk factors for withdrawal. RESULTS: Twenty-nine patients completed the study. They received lorazepam for a median duration of about 21 days, and withdrawal occurred in 7 patients. There were no significant differences in demographic variables, lorazepam dosage or other sedative therapy, or lorazepam serum concentrations between patients with withdrawal and those without withdrawal. No predictors of withdrawal were identified. CONCLUSIONS: Withdrawal occurred in 24% of critically ill children receiving long-term sedation from lorazepam. Risk factors for withdrawal are unknown. PMID- 16720708 TI - Vancomycin-induced elevation of liver enzyme levels. AB - OBJECTIVE: To report a case of oral vancomycin-induced elevation of liver enzyme levels. CASE SUMMARY: A 57-year-old man with multiple medical conditions requiring systemic antibiotic therapy developed numerous Clostridium difficile associated enterocolitis episodes. The patient did not respond adequately to oral metronidazole, as evidenced by his continuing diarrhea. He was treated with oral vancomycin on 5 separate occasions (with doses from 125 to 500 mg/day), each of which resulted in significant elevations in alanine aminotransferase (to 371 U/L) and aspartate aminotransferase (to 203 U/L) levels. The elevations resolved on each occasion with discontinuation of vancomycin. DISCUSSION: Vancomycin, a glycopeptide antibiotic, has primary activity against gram-positive bacteria. Oral vancomycin can be used for the treatment of C. difficile-associated enterocolitis in patients who fail to respond to or are intolerant to metronidazole therapy. Oral vancomycin has very poor bioavailability and, as of May 4, 2006, has not been associated with hepatic toxicity. Inflammatory bowel disease processes can result in increased absorption of oral vancomycin. CONCLUSIONS: This is the first reported case of oral vancomycin-induced elevation of hepatic enzyme levels. Use of the Naranjo probability scale indicated that this was a probable adverse drug-associated event. PMID- 16720709 TI - Coxibs versus combination NSAID and PPI therapy for chronic pain: an exploration of the risks, benefits, and costs. AB - OBJECTIVE: To systematically review studies qualitatively to compare the risks (gastrointestinal [GI] and cardiovascular) and benefits (pain control) of cyclooxygenase-2 inhibitors (coxibs) relative to an alternative therapy of a nonselective nonsteroidal antiinflammatory drug (NSAID) combined with a proton pump inhibitor (PPI) and explore circumstances when coxibs may be appropriate. METHODS: Relevant studies were identified through a search of MEDLINE (Ovid Technologies, 1985-November 2005; English language, clinical trial), PubMed (1985 November 2005; English language, clinical trial, humans), and the Cochrane Collaboration using the terms selective COX-2 inhibitors and coxibs, as well as the various chemical names for specific coxib agents. Studies that compared a coxib with a nonselective NSAID and provided data concerning our outcomes of interest were included and categorized by the outcome variable, as well as by the specific coxib studied. RESULTS: The majority of the numerous studies that evaluated pain as an endpoint showed no difference between coxib and nonselective NSAID therapy. However, while limited, preliminary safety data regarding the effects of both classes on the upper and lower GI tract suggest coxib superiority. Although coxibs are associated with an increased risk of cardiovascular adverse events (CVEs) compared with placebo, this effect has not been conclusively shown compared with nonselective NSAIDs. Currently, coxib therapy is more expensive than combination therapy using a nonselective NSAID plus a PPI. CONCLUSIONS: Compared with combination therapy including a nonselective NSAID and PPI, coxibs provide equivalent pain control and may have a lower GI tract complication profile, but at an unknown increased risk of CVEs and a greater financial cost. Coxib therapy may be an appropriate treatment for chronic pain in select patients with higher risks of GI complications, lower risk of CVEs, and in whom greater cost is not a restraint. PMID- 16720710 TI - Steroid therapy for a case of severe drug-induced cholestasis. AB - OBJECTIVE: To report a severe case of cholestatic liver disease successfully treated with corticosteroids following combined therapy with clarithromycin and nimesulide. CASE SUMMARY: A 15-year-old girl was admitted with cholestasis probably related to treatment with clarithromycin and nimesulide for an upper respiratory tract infection. Other causes of liver disease (infections, metabolic liver disorders, genetic cholestatic syndromes, autoimmune diseases, primary biliary tract disorders) were excluded. Liver biopsy showed a severe canalicular cholestasis with bile plugs in dilated bile canaliculi, giant cell transformation, and portal and lobular infiltrate. An objective causality assessment suggested that cholestasis was probably related to clarithromycin and/or nimesulide use. No benefit was derived from a course of ursodeoxycholic acid therapy. Since the patient experienced a progressive worsening in cholestasis, prednisone was started after 20 days. This therapy was promptly followed by improvement in clinical and laboratory test results. After 2 months of prednisone treatment, the patient became symptom-free with normal liver function tests. DISCUSSION: The manifestations of drug-induced hepatotoxicity are highly variable, ranging from asymptomatic hypertransaminemia to fulminant hepatic failure. No specific treatment for drug-induced hepatotoxicity exists. Early recognition and drug withdrawal are the keys to management of hepatotoxicity, but in some cases, liver disease may persist despite discontinuation of the drug. Possible advantages of corticosteroid therapy have not been well demonstrated. CONCLUSIONS: Application of the Naranjo probability scale indicates a probable relationship between cholestasis and nimesulide plus clarithromycin use. This case draws attention to a possible therapeutic option for some cases of drug-induced hepatotoxicity that show a severe course without any sign of improvement. PMID- 16720711 TI - Death or survival--progesterone-dependent cell fate decisions in the human endometrial stroma. AB - The human endometrium undergoes cyclical waves of proliferation, differentiation and apoptosis in response to the rise and fall in ovarian oestradiol and progesterone levels. These hormonal responses in endometrial cells must be tightly kept in check to safeguard tissue homeostasis throughout reproductive life. The discovery that differentiating endometrium highly expresses the tumour suppressor p53, the forkhead transcription factor FOXO1, and promyelocytic leukaemia zinc finger protein (PLZF) has provided new insights into the molecular basis of life and death decisions in response to sex steroid hormones. PMID- 16720712 TI - FOXL2 activates P450 aromatase gene transcription: towards a better characterization of the early steps of mammalian ovarian development. AB - Previous studies have equated FOXL2 as a crucial actor in the ovarian differentiation process in different vertebrate species. Its transcriptional extinction in the polled intersex syndrome (PIS) leads primarily to a drastic decrease of aromatase (CYP19) expression in the first steps of goat ovarian development. In this study, we provide a better characterization of early ovarian development in goat, and we provide experimental evidence demonstrating that FOXL2 represents a direct transcriptional activator of the CYP19 gene through its ovarian-specific promoter 2. Moreover, the ovarian location of FOXL2 and CYP19 proteins, together with their expression profiles in the female gonads, stress the involvement of FOXL2 co-factor(s) for regulating CYP19 transcription. Expressional analyses show that activin-betaA can be considered as a strong candidate for being one of these FOXL2 co-factors. Finally, we discuss evidence for a role of activin and estrogens in somatic and germinal cell proliferation occurring before germ cell meiosis. This period, of 20 days in goat, seems to have no equivalent in mouse. This species-specific difference could explain the phenotype discrepancy observed between XX goat PIS(-/-) and XX mouse Foxl2(-/-). PMID- 16720713 TI - Cycling and early pregnant endometrium as a site of regulated expression of the vitamin D system. AB - In addition to its calciotropic function, the secosteroid 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)), has potent anti-proliferative/immunomodulatory effects on various tissues. Consistently, the enzyme that catalyzes the synthesis of 1,25(OH)(2)D(3), 1alpha-hydroxylase (1alpha-OHase) and the vitamin D receptor have a widespread tissue distribution. Among site-specific functions, the hormone has been suggested to be involved in uterine physiology. However, molecular analysis of the vitamin D system in normal endometrium throughout the menstrual cycle as well as its regulation in the context of endometrial physiological and pathological events have received very limited attention. Thus, we have studied expression, localization and regulation of 1alpha-OHase in human cycling and early pregnant endometrium. The capacity for 1alpha-hydroxylation and the presence of vitamin D receptor in endometrial cells have also been evaluated. The functional significance of these findings has been tested by evaluating gene expression of the catabolic enzyme, vitamin D 24-hydroxylase, and of the adhesion protein, osteopontin. Finally, to verify any potential dysfunction of the vitamin D system in endometriosis, a reproductive disease characterized by immune mediated anomalies, we have analyzed expression of 1alpha-OHase in both eutopic and ectopic endometrium of affected patients. Results obtained showed that the active form of the 1alpha-OHase gene was expressed in human endometrial stromal cells independent of the cycle phase but with a significant increase in early pregnant decidua. A similar profile was observed for the protein, which was abundantly expressed in the cytoplasm of both endometrial stroma and epithelial glands. Both cycling and early pregnant endometrial cells also expressed the vitamin D receptor. In the same cells, 1alpha-OHase mRNA levels were significantly stimulated by the pro-inflammatory cytokine interleukin (IL)-1beta (50 and 500 pg/ml) while addition of the active form of the hormone could modulate both CYP24 and osteopontin gene expression. The 1alpha-OHase gene was also expressed in ectopic endometrium and its levels were increased in proliferative phase cultures derived from patients with endometriosis. Human cycling endometrium may be included among the extrarenal sites able to synthesize vitamin D. The IL-1beta-mediated induction of 1alpha-OHase gene and the hormonal modulation of osteopontin support a role for the hormone in the immunological mechanisms underlying uterine function. Abnormalities of this system are present in endometriosis. PMID- 16720714 TI - Pleiotropic effects of the steroid hormone 1,25-dihydroxyvitamin D3 on the recruitment of mesenchymal lineage progenitors in fetal rat calvaria cell populations. AB - The steroid hormone 1,25-dihydroxyvitamin D3 (1,25(OH)(2)D(3)) inhibits osteogenesis while stimulating adipogenesis in vitro. We hypothesized that 1,25(OH)(2)D(3) redirects the fate of osteoblast/adipocyte bipotential progenitors and other potential progenitors towards adipogenesis, a process possibly underlying the pathogenesis of osteopenic diseases such as osteoporosis. We therefore tested the global effects of 1,25(OH)(2)D(3) on the recruitment of mesenchymal progenitors including osteogenic, chondrogenic, adipogenic and myogenic lineages (colony forming cell (CFC)-osteoblast (CFC-O), CFC-chondrocyte (CFC-C), CFC-adipocyte (CFC-A), and CFC-myoblast (CFC-M) respectively) in rat calvaria (RC) cell populations using gene expression profiling of single cell derived colonies. Based on expression of lineage specific transcripts, 86% of single cell-derived colonies in untreated cultures simultaneously co-expressed transcripts of two, three, or four of the mesenchymal lineages tested. The distribution of mesenchymal progenitors in 1,25(OH)(2)D(3)-treated cultures was significantly changed compared with the control group, i.e. CFC-O were reduced (from 6 to 0%) and CFC-O/A bipotential (0 to 8.2%), CFC-C (4 to 10.2%) and CFC Fibroblast (CFC-F) (4 to 16%) were increased. 1,25(OH)(2)D(3) did not affect the frequency of tri- or tetra-lineage colonies. Single lineage CFC-A colonies were not detected in either the control or 1,25(OH)(2)D(3) treatment group under the conditions tested. Since the parietal bones used for cell isolation derive from neuroectoderm, we also analyzed for expression of the neural markers nestin and beta3 tubulin in these colonies. Surprisingly, 90% (45 of 50) of the colonies in the control group expressed neural markers, a frequency not changed by 1,25(OH)(2)D(3) treatment. The current studies demonstrate the global and developmental stage-specific effects of 1,25(OH)(2)D(3) on mesenchymal lineage progenitors, and suggest that the effects of 1,25(OH)(2)D(3) on osteogenesis and adipogenesis in RC populations are mediated, at least in part, by increased recruitment of CFC-O/A, but not CFC-A type precursors. PMID- 16720715 TI - Prolactin inhibits cell loss and decreases matrix metalloproteinase expression in the involuting mouse mammary gland but fails to prevent cell loss in the mammary glands of mice expressing IGFBP-5 as a mammary transgene. AB - Insulin-like growth factor-binding protein 5 (IGFBP-5) mediates involution of the mammary gland. The decrease in DNA content and mammary gland weight which accompanies involution was inhibited by prolactin (PRL) in wild-type but not transgenic mice expressing IGFBP-5. Phospho-STAT5 protein levels were significantly lower in IGFBP-5 transgenic mice during lactation suggesting that IGFBP-5 antagonises PRL signalling in the mammary epithelium. In contrast, phospho-STAT3 levels increased during involution to a similar extent in both wild type and transgenic mice and were unaffected by PRL. PRL inhibited gene expression of matrix metalloproteinases (MMPs) 3 and 12 but not tissue plasminogen activator or plasmin in wild-type and transgenic animals. The effects of PRL on MMPs appear to be indirect since PRL failed to inhibit MMP-3, -7 or -12 expression in HC-11 cells or in a co-transfection including an activated PRL receptor, STAT5 and a MMP-3-luciferase reporter gene. PRL is a potent inhibitor, both of cell death, an effect which is suppressed by IGFBP-5, and of MMP expression, which is independent of the actions of IGFBP-5. PMID- 16720716 TI - Human chorionic gonadotropin-dependent induction of an equine aldo-keto reductase (AKR1C23) with 20alpha-hydroxysteroid dehydrogenase activity during follicular luteinization in vivo. AB - Aldo-keto reductases (AKRs) are multifunctional enzymes capable of acting on a wide variety of substrates, including sex steroids. AKRs having 20alpha hydroxysteroid dehydrogenase (20alpha-HSD) activity can reduce progesterone to 20alpha-hydroxy-4-pregnen-3-one (20alpha-DHP), a metabolite with lower affinity for the progesterone receptor. The objective of this study was to investigate the regulation of equine AKR1C23 during human chorionic gonadotropin (hCG)-induced ovulation/luteinization. The equine AKR1C23 cDNA was cloned and shown to encode a 322 amino acid protein that is conserved (71-81% identity) when compared with mammalian orthologs. RT-PCR/Southern blotting analyses were performed to study the regulation of AKR1C23 transcripts in equine preovulatory follicles isolated between 0 and 39 h after hCG treatment (ovulation occurring 39-42 h post-hCG). Results showed the presence of low AKR1C23 expression before hCG treatment, but a marked increase was observed in follicles obtained 12 h after hCG (P<0.05). Analyses of isolated preparations of granulosa and theca interna cells identified low mRNA expression in both cell types prior to hCG treatment, with granulosa cells clearly being the predominant site of follicular AKR1C23 mRNA induction. A specific polyclonal antibody was raised against a fragment of the equine protein and immunoblotting analyses showed an increase in AKR1C23 protein in granulosa cell extracts when comparing follicles isolated at 36 h post-hCG vs those collected prior to treatment, in keeping with mRNA results. Immunohistochemical data confirmed the induction of the enzyme in follicular cells after hCG treatment. The enzyme was tested for 20alpha-HSD activity and was shown to exhibit a K(M) of 3.12 microM, and a V(max) of 0.86 pmol/min per 10 microg protein towards progesterone. The levels of 20alpha-DHP measured in follicular fluid reflected this activity. Collectively, these results demonstrate for the first time that the gonadotropin-dependent induction of follicular luteinization is accompanied by an increase in AKR1C23 expression. Considering the 20alpha-HSD activity of AKR1C23, its regulated expression in luteinizing preovulatory follicles may provide a biochemical basis for the increase in ovarian 20alpha-DHP observed during gonadotropin-induced luteinization/ovulation. (The nucleotide sequence reported in this paper has been submitted to GenBank with accession number AY955082.). PMID- 16720717 TI - Androgen receptor and prostate apoptosis response factor-4 target the c-FLIP gene to determine survival and apoptosis in the prostate gland. AB - Androgen receptor (AR) is a ligand-activated transcription factor that mediates the action of androgens and is essential for the growth, function, and cell differentiation of the prostate gland. Here, we demonstrated that the prostate apoptosis response factor-4 (par-4) functions as a novel AR coactivator. Par-4 physically interacted with the DNA-binding domain of AR, enhanced AR interaction with DNA, and increased AR-dependent transcription. Par-4 enhanced the c-FLIP promoter activity and was recruited on to the c-FLIP gene in the presence of androgens, and the dominant-negative par-4 decreased c-FLIP expression. These results suggest that, in addition to its proapoptotic function, par-4 acts as a novel transcription cofactor for AR to target c-FLIP gene expression. In addition, we demonstrated that loss of c-FLIP expression was essential for castration-induced apoptosis in the prostate gland and that enhanced c-FLIP expression was associated with prostate cancer progression to the androgen resistant stage. Our data shed light on a transcription-mediated mechanism for the effects of the AR pathway on cell survival and apoptosis. PMID- 16720718 TI - Defining high-fat-diet rat models: metabolic and molecular effects of different fat types. AB - High-fat (HF)-diet rodent models have contributed significantly to the analysis of the pathophysiology of the insulin resistance syndrome, but their phenotype varies distinctly between different studies. Here, we have systematically compared the metabolic and molecular effects of different HF with varying fatty acid compositions. Male Wistar rats were fed HF diets (42% energy; fat sources: HF-L - lard; HF-O - olive oil; HF-C - coconut fat; HF-F - fish oil). Weight, food intake, whole-body insulin tolerance and plasma parameters of glucose and lipid metabolism were measured during a 12-week diet course. Liver histologies and hepatic gene expression profiles, using Affymetrix GeneChips, were obtained. HF-L and HF-O fed rats showed the most pronounced obesity and insulin resistance; insulin sensitivity in HF-C and HF-F was close to normal. Plasma omega-3 polyunsaturated fatty acid (omega-3-PUFA) and saturated fatty acid (C(12)-C(14), SFA) levels were elevated in HF-F and HF-C animals respectively. The liver histologies showed hepatic steatosis in HF-L, HF-O and HF-C without major inflammation. Hepatic SREBP1c-dependent genes were upregulated in these diets, whereas PPARalpha-dependent genes were predominantly upregulated in HF-F fed rats. We detected classical HF effects only in diets based on lard and olive oil (mainly long-chain, saturated (LC-SFA) and monounsaturated fatty acids (MUFA)). PUFA- or MC-SFA-rich diets did not induce insulin resistance. Diets based on LC SFA and MUFA induced hepatic steatosis with SREBP1c activation. This points to an intact transcriptional hepatic insulin effect despite resistance to insulin's metabolic actions. PMID- 16720719 TI - Antibody inhibition of synaptosomal protein of 25 kDa (SNAP-25) and syntaxin 1 reduces rapid exocytosis in insulin-secreting cells. AB - SNARE-proteins (soluble NSF-attachment protein receptor) are important for Ca(2+) dependent exocytosis. We have used capacitance measurements and confocal imaging to dissect the role of synaptosomal protein of 25 kDa (SNAP-25) and syntaxin 1 in rapid exocytosis in insulin-secreting pancreatic beta-cells. Following immunoneutralization of syntaxin 1 and SNAP-25, exocytosis was strongly reduced and associated with a marked reduction in the size of the readily releasable pool (RRP) by 65% and 86% in the presence of the anti-SNAP-25 and anti-syntaxin 1 antibodies respectively. The size of the immediately releasable pool (IRP), a subset of RRP in close association with the voltage-dependent Ca(2+)-channels, was reduced to an equal extent. The reduction in IRP correlated with slowed release kinetics and the time constant (tau) increased from a control value of 16 to 36 ms and 51 ms after inclusion of anti-SNAP-25 and anti-syntaxin 1 antibodies respectively in the pipette solution. We further show that SNAP-25 and syntaxin 1 aggregate in clusters along the plasma membrane. The size of these clusters was estimated to be approximately 300 nm and every beta-cell contained approximately 400 SNAP-25/syntaxin 1 clusters. Whereas the inhibitory action of the anti syntaxin 1 antibody on exocytosis could be attributed almost entirely to suppression of the voltage-dependent Ca(2+)-current (-40%), the effect of the anti-SNAP-25 antibody was not mediated by decreased Ca(2+)-entry and is more likely due to a direct interference with the exocytotic machinery. Our data are consistent with the concept that both syntaxin 1 and SNAP-25 are required for rapid exocytosis in beta-cells. PMID- 16720720 TI - Thyroid hormone-mediated negative transcriptional regulation of Necdin expression. AB - Unliganded thyroid hormone receptors (apoTRs) repress transcription of hormone activated genes by recruiting corepressors to the promoters. In contrast, on promoters containing so-called negative thyroid hormone response elements (nTREs), apoTRs activate transcription. A number of different molecular mechanisms have been described as to how apoTRs activate transcription varying with the target gene of the study. Here we demonstrate that thyroid hormone regulates the transcription of the Necdin gene, a developmentally regulated candidate gene for the genomic imprinting-associated neurobehavioural disorder, Prader-Willi syndrome. ApoTRs activate Necdin expression through an nTRE in its promoter, downstream of the transcription start site. The nTRE of the Necdin gene resembles the nTREs of the TSHbeta genes of the hypothalamus-pituitary-thyroid axis in the sequence, position in the promoter, and mode of activation. We show that this group of nTRE-driven genes shares the requirements for binding of the retinoic X receptor and nuclear receptor corepressor/silencing mediator of retinoid and thyroid hormone receptors (NCoR/SMRT) for full ligand-independent activation, whereas there is no need for association of the p160 family of coactivators. In accordance with the requirement for corepressors, Necdin expression is influenced by deacetylase activity, suggesting that histone deacetylases and corepressors as well could function as activators of transcription, depending on the promoter context. PMID- 16720721 TI - CCAAT/enhancer-binding protein delta activates the Runx2-mediated transcription of mouse osteocalcin II promoter. AB - CCAAT/enhancer-binding proteins (C/EBPs) are involved in the regulation of cell proliferation, differentiation, and control of metabolic function. Although the roles of C/EBPs in osteoblasts are largely unknown, both C/EBPbeta and -delta have been shown to enhance rat osteocalcin promoter activity through the synergistic activation of Runx2 at the C/EBP element. Here we show that in the mouse, C/EBPdelta increases the expression of osteocalcin whereas C/EBPbeta does not. This increased expression was found to occur at the transcriptional level, as demonstrated by the increased transcriptional activity from mouse osteocalcin II (OG2) promoter by C/EBPdelta. Although we found three putative C/EBP sites in the -637/+/-34 region of the OG2 promoter, none of these sites showed binding activity with in vitro translated C/EBP proteins. Notably, we show that C/EBPdelta physically interacts with Runx2 and that C/EBPdelta overexpression increases binding between the Runx2-C/EBPdelta complex and the OSE2 element, a critical osteoblast-specific cis-acting element in the OG2 promoter. Consistent with these DNA binding data, a mutation in OSE2 abrogated the stimulatory effect of C/EBPdelta on this promoter activity. Finally, chromatin immunoprecipitation analysis in MC3T3-E1 cells showed in vivo occupancy of the OG2 promoter by Runx2 and C/EBPdelta. In conclusion, C/EBPdelta was found to regulate mouse osteocalcin OG2 promoter activity indirectly by interacting with Runx2 in the context of the OSE2 element and this subsequently resulted in the cooperative activation of the OG2 promoter. PMID- 16720722 TI - Cortistatin mimics somatostatin by inducing a dual, dose-dependent stimulatory and inhibitory effect on growth hormone secretion in somatotropes. AB - Cortistatin is a recently discovered neuropeptide that is structurally related to somatostatin, the classic inhibitor of growth hormone (GH) release. Cortistatin binds with high affinity to all five somatostatin receptors (sst1-5), and, like somatostatin, cortistatin inhibits in vivo GH release in man and rats. In this report, we compared the in vitro actions of cortistatin and somatostatin using primary pig pituitary cell cultures. In this species, we have previously reported that somatostatin not only inhibits GH-releasing hormone (GHRH)-stimulated GH release at high doses, but also stimulates basal GH release at low (pM) doses, a dual response that is markedly dependent on the subpopulation of pituitary somatotropes examined. Results reported herein demonstrate that cortistatin closely mimics the dose-dependent inhibitory and stimulatory effects of somatostatin on GH secretion. As cortistatin, unlike somatostatin, binds to the human receptor for ghrelin/GH secretagogs (GHS-R), we also investigated whether cortistatin stimulates GH release through this receptor by using a synthetic, short form of cortistatin, cortistatin-8 (CST8), which lacks the sst-binding capacity of full-length cortistatin but retains its GHS-R-binding capacity. Interestingly, CST8 stimulated GH release only at low doses (10(-15) M), and did not reduce GH secretion stimulated by GHRH, ghrelin, or low-dose, full-length cortistatin, yet it counteracted that induced by a nonpeptidyl GHS, L-163 255. Taken together, our results indicate that the dual, inhibitory and stimulatory effects of cortistatin on GH release closely parallel those of somatostatin and are probably mediated by the same receptor(s) and signaling pathway(s) for both peptides. Furthermore, they suggest that the pathway(s) activated by cortistatin (and somatostatin) to stimulate GH release are not initiated by GHS-R activation. PMID- 16720723 TI - GATA-4 is a granulosa cell factor employed in inhibin-alpha activation by the TGF beta pathway. AB - Part of heterodimeric inhibin, inhibin-alpha is crucial for mammalian ovarian function. Regulation of inhibin-alpha expression in granulosa cells is both endocrine, primarily by follicle-stimulating hormone (FSH), and paracrine, primarily by members of the transforming growth factor beta (TGF-beta) superfamily. Smad proteins transmit TGF-beta signals to the nucleus, but the cooperating transcription factors involved in inhibin-alpha promoter activation remain unknown. Transcription factor GATA-4 regulates inhibin-alpha in gonadal cells, and the FSH cascade activates GATA-4. We hypothesized that the TGF-beta signalling cascade and GATA-4 also cooperate to regulate inhibin-alpha expression. In KK-1 granulosa tumour cells, which resemble normal granulosa cells and express inhibin-alpha, we found that TGF-beta upregulated GATA-4 expression. Transient transfection experiments in KK-1 cells demonstrated that dominant negative GATA-4 variants or mutations of GATA-binding sites in the inhibin-alpha promoter attenuated TGF-beta-induced gene activation. In GATA-4-deficient COS-7 cells, TGF-beta enhanced the expression of the inhibin-alpha promoter only in the presence of exogenous GATA-4. Smad3, but not Smad2, cooperated with GATA-4 in the transcriptional activation of the inhibin-alpha promoter, and immunoprecipitation experiments in KK-1 cells revealed a physical Smad3:GATA-4 interaction. Our data suggest that GATA-4, interacting with Smad3, is a cofactor for TGF-beta signalling to activate inhibin-alpha in granulosa cells. PMID- 16720724 TI - FKBP12 functions as an adaptor of the Smad7-Smurf1 complex on activin type I receptor. AB - The cytoplasmic immunophilin FKBP12, a 12 kDa FK506-binding protein, has been shown to act as an inhibitor for transforming growth factor-beta (TGF-beta) signaling. FKBP12 binds to the glycine- and serine-rich motif (GS motif) of the TGF-beta type I receptor, and functions as a secure switch to prevent the leaky signal. Upon stimulation with ligand, FKBP12 is released from the receptor to fully propagate the signal. We found that activin, a member of TGF-beta superfamily, also induced the dissociation of FKBP12 from the activin type I receptor (ALK4). However, we observed that the released FKBP12 associates again with the receptor a few hours later. FKBP12 also interacted with another inhibitory molecule of activin signal, Smad7, in an activin-dependent manner, and formed a complex with Smad7 on the type I receptor. FK506, a chemical ligand for FKBP12, which dissociates FKBP12 from the receptor, decreased the interaction between Smad7 and Smad ubiquitin regulatory factor 1 (Smurf1). FK506 also inhibited the ubiquitination of the type I receptor by Smurf1. These findings indicate a new inhibitory function of FKBP12 as an adaptor molecule for the Smad7 Smurf1 complex to regulate the duration of the activin signal. PMID- 16720725 TI - Vasopressin increases GAGA binding activity to the V1b receptor promoter through transactivation of the MAP kinase pathway. AB - Previous studies show that binding of nuclear proteins to GAGA repeats (GAGA box) in the vasopressin type 1b receptor (V1bR) promoter is essential for transcriptional initiation of the gene. To determine whether increased vasopressin (VP) during stress activates V1bR expression through the GAGA box, we examined the effects of VP on GAGA binding activity and on the ability of the V1bR promoter to recruit RNA polymerase in the hypothalamic cell line, H32. In chromatin immunoprecipitation assays, VP induced RNA polymerase II recruitment by the wild type V1bR promoter but not by a construct with the major GAGA box deletion. VP (10 min) also increased binding of nuclear proteins to radiolabeled GAGA oligonucleotides in electromobility shift assays. VP-induced GAGA binding activity was potentiated by the protein kinase C inhibitor, calphostin C, and was prevented by the MEK inhibitor, UO126, and the epidermal growth factor receptor (EGFR) inhibitor, AG1478, suggesting that VP activates GAGA binding through transactivation of the EGFR. This was confirmed by western blot experiments showing rapid increases in phospho ERK after incubation with VP, an effect that was potentiated by calphostin C and inhibited by UO12 and AG1478, as well as by the ability of VP to phosphorylate the EGFR. Using receptor selective VP analogs we showed that both V1aR and V1bR subtypes can mediate GAGA binding activation in H32 cells. This study demonstrates that VP stimulates GAGA binding to the V1bR promoter through transactivation of the EGFR and MAP kinase. The data support the hypothesis that VP contributes to pituitary V1bR upregulation during stress through GAGA binding-mediated transcriptional activation. PMID- 16720726 TI - Cell-specificity of transforming growth factor-beta response is dictated by receptor bioavailability. AB - Members of the transforming growth factor-beta (TGFbeta) family control diverse cellular responses including differentiation, proliferation, controlled cell death and migration. The response of a cell to an individual ligand is highly restricted yet the signaling pathways for TGFbeta, activin and bone morphogenic proteins share a limited number of receptors and activate similar intracellular cytoplasmic co-regulators, Smads. A central question in the study of this family of ligands is how cells titrate and integrate each TGFbeta-like signal in order to respond in a cell- and ligand-specific manner. This study uses the pituitary gonadotrope cell line, LbetaT2, as a model to delineate the relative contribution of TGFbeta and activin ligands to follicle-stimulating hormone (FSH) biosynthesis. It was found that pituitary gonadotrope cells do not express the TGFbeta type II (TbetaRII) receptor and are therefore not responsive to the TGFbeta ligand. Transfection of the receptor restores TGFbeta signaling capabilities and the TGFbeta-mediated stimulation of FSHbeta gene transcription in LbetaT2 cells. Consequently, we evaluated the presence of the TbetaRII in the adult mouse pituitary. TbetaRII does not co-localize with FSH-producing cells; however it is detected on the cell surface of prolactin- and growth hormone positive cells. Taken together, these results suggest that the bioavailability of the TGFbeta-specific receptor rather than TGFbeta dictates pituitary gonadotrope selectivity to activin, which is necessary to maintain normal reproductive function. It is likely that the ligand-restricted mechanisms employed by the gonadotrope are present in other cells, which could explain the distinct control of many cellular processes by members of the TGFbeta superfamily. PMID- 16720727 TI - Functional characterization and kinetic studies of an ancestral lamprey GnRH-III selective type II GnRH receptor from the sea lamprey, Petromyzon marinus. AB - The recently cloned lamprey GnRH receptor was shown to have several unique features, including the longest intracellular C-terminal tail (120 amino acids (aa)) of any previously described GnRH receptor. In the current study, a series of experiments were performed examining cAMP responses, binding kinetics, whole cell competitive binding assays and internalization studies of the lamprey GnRH receptor using a series of three C-terminal tail truncations (80 aa, 40 aa and 0 aa) to better describe the functional significance of this unique vertebrate GnRH receptor. Activation of the lamprey GnRH receptor was shown to stimulate cAMP production in a dose-dependent manner when treated with either lamprey GnRH-I (LogEC50 -6.57+/-0.15) or lamprey GnRH-III (LogEC(50) -8.29+/-0.09). Truncation analysis indicated that the membrane proximal 40 aa of the lamprey GnRH receptor C-terminal tail contain a motif required for cAMP accumulation. Saturation binding assays using the wild type and truncated lamprey GnRH receptors revealed that all of three truncated lamprey GnRH receptors were capable of binding lamprey GnRH-I. Competitive, intact cell-binding assays suggested that the lamprey GnRH receptor is lamprey GnRH-III selective, based on the observed pharmacological profile: lamprey GnRH-III (Inhibitory constant (Ki) 0.708+/-0.245 nM)=chicken GnRH-II (Ki 0.765+/-0.160 nM) > mammalian GnRH (Ki 12.9+/-1.96 nM) > dAla(6)Pro(9)NEt mammalian GnRH (Ki 21.6+/-9.68 nM) > lamprey GnRH-I (Ki 118.0+/ 23.6). Finally, the lamprey GnRH receptor was shown to undergo rapid ligand dependent internalization, which was significantly diminished in the tail-less truncated form. We have shown from our current and our previous structural studies that this unique lamprey GnRH receptor shares several characteristics of both type I and type II GnRH receptors which suggests that this receptor has retained ancestral characteristics that can provide insight into the function and evolution of the vertebrate GnRH receptor family. PMID- 16720728 TI - Mechanism of nitric oxide action on inhibitory GABAergic signaling within the nucleus tractus solitarii. AB - The cellular mechanisms mediating nitric oxide (NO) modulation of the inhibitory transmission in the nucleus tractus solitarii (NTS) remain unclear, even though this could be extremely important for various physiological and pathological processes. Specifically, in the NTS NO-evoked glutamate and gamma-aminobutyric acid (GABA) release might contribute to pathological hypertension. In cultured rat brainstem slices, NTS GABAergic neurons were targeted using an adenoviral vector to express enhanced green fluorescent protein and studied with a combination of patch clamp and confocal microscopy. Low nanomolar concentrations of NO increased intracellular Ca2+ concentration ([Ca2+]i) in somata, dendrites, and putative axons of GABAergic neurons, with axons being the most sensitive compartment. This effect was cGMP mediated and not related to depolarization or indirect presynaptic effects on glutamatergic transmission. Blockade of the cyclic adenosine diphosphate ribose (cADPR)/ryanodine-sensitive stores but not the inositol triphosphate-sensitive stores, inhibited NO effect. Since cADPR/ryanodine-sensitive stores are implicated in the Ca2+-induced Ca2+ release, NO can be expected to potentiate GABA release. In support of this notion, a cADPR antagonist abolished the NO-induced potentiation of GABAergic inhibitory postsynaptic potentials in the NTS. Thus, the NO-cGMP-cADPR-Ca2+ pathway, previously described in sea urchin eggs, also operates in mammalian GABAergic neurons. Potentiation of GABA release by NO may have implications for numerous brain functions. PMID- 16720730 TI - Maspin is physically associated with [beta]1 integrin regulating cell adhesion in mammary epithelial cells. AB - Maspin is a tumor-suppressor serpin (serine protease inhibitor), which inhibits cell invasion and migration. Here, we analyzed maspin function in cell adhesion in nontransformed mammary epithelial cells and investigated the underlying mechanism involved in this process. We report that maspin acts in the early steps in the cell adhesion process. Addition of recombinant maspin rapidly increased MCF-10A cell adhesion to the endogenously deposited matrix, and conversely both an antimaspin antibody (Ab) and maspin knockdown by RNA interference resulted in decreased cell adhesion. Mutation analyses revealed that a region of 86 amino acids located between aa 139 and aa 225 was responsible for maspin effect on adhesion. In addition, we show that maspin is associated with detergent-insoluble cortical cytoskeleton elements. Collectively, these results suggest that maspin is part of the supramolecular structure of the adhesion plaque and it modulates cell adhesion via a beta1 integrin-dependent mechanism. PMID- 16720729 TI - Host Lewis phenotype-dependent Helicobacter pylori Lewis antigen expression in rhesus monkeys. AB - Both human and H. pylori populations are polymorphic for the expression of Lewis antigens. Using an experimental H. pylori challenge of rhesus monkeys of differing Lewis phenotypes, we aimed to determine whether H. pylori populations adapt their Lewis phenotypes to those of their hosts. After inoculation of four monkeys with a mixture of seven strains identified by RAPD-polymerase chain reaction, H. pylori Lewis expression was followed in 86 isolates obtained over 40 wk. Host Lewis(a/b) secretion status was characterized by immunological assays. Fingerprints of the predominating strain (J166) were identical in all four animals after 40 wk, but its Lewis phenotype had substantial variability in individual hosts. At 40 wk, J166 populations from two Lewis(a-b+) animals predominantly expressed Lewis(y). In contrast, J166 populations had switched to a Lewis(x) dominant phenotype in the two Lewis(a+b-) animals; a frame shift in futC, regulating conversion of Lewis(x) to Lewis(y), accounted for the phenotypic switch. The results indicate that individual cells in H. pylori populations can change Lewis phenotypes during long-term colonization of natural hosts to resemble those of their hosts, providing evidence for host selection for bacterial phenotypes. PMID- 16720731 TI - Estrogenic effects of natural and synthetic compounds including tibolone assessed in Saccharomyces cerevisiae expressing the human estrogen alpha and beta receptors. AB - The human estrogen receptors (hER)alpha and hERbeta, differentially expressed and localized in various tissues and cell types, mediate transcriptional activation of target genes. These encode a variety of physiological reproductive and nonreproductive functions involved in energy metabolism, salt balance, immune system, development, and differentiation. As a step toward developing a screening assay for the use in applications where significant numbers of compounds or complex matrices need to be tested for (anti) estrogenic bioactivity, hERalpha and hERbeta were expressed in a genetically modified Saccharomyces cerevisiae strain, devoid of three endogenous xenobiotic transporters (PDR5, SNQ2, and YOR1). By using receptor-mediated transcriptional activation of the green fluorescent protein optimized for expression in yeast (yEGFP) as reporter 17 natural, comprising estrogens and phytoestrogens or synthetic compounds among which tibolone with its metabolites, gestagens, and antiestrogens were investigated. The reporter assay deployed a simple and robust protocol for the rapid detection of estrogenic effects within a 96-well microplate format. Results were expressed as effective concentrations (EC50) and correlated to other yeast based and cell line assays. Tibolone and its metabolites exerted clear estrogenic effects, though considerably less potent than all other natural and synthetic compounds. For the blood serum of two volunteers, considerable higher total estrogenic bioactivity than single estradiol concentrations as determined by immunoassay was found. Visualization of a hERalpha/GFP fusion protein in yeast revealed a sub cellular cytosolic localization. This study demonstrates the versatility of (anti) estrogenic bioactivity determination using sensitized S. cerevisiae cells to assess estrogenic exposure and effects. PMID- 16720732 TI - Epithelial cell PPAR[gamma] contributes to normal lung maturation. AB - Peroxisome proliferator-activated receptor (PPAR)-gamma is a member of the nuclear hormone receptor superfamily that can promote cellular differentiation and organ development. PPARgamma expression has been reported in a number of pulmonary cell types, including inflammatory, mesenchymal, and epithelial cells. We find that PPARgamma is prominently expressed in the airway epithelium in the mouse lung. In an effort to define the physiological role of PPARgamma within the lung, we have ablated PPARgamma using a novel line of mice capable of specifically targeting the airway epithelium. Airway epithelial cell PPARgamma targeted mice display enlarged airspaces resulting from insufficient postnatal lung maturation. The increase in airspace size is accompanied by alterations in lung physiology, including increased lung volumes and decreased tissue resistance. Genome-wide expression profiling reveals a reduction in structural extracellular matrix (ECM) gene expression in conditionally targeted mice, suggesting a disruption in epithelial-mesenchymal interactions necessary for the establishment of normal lung structure. Expression profiling of airway epithelial cells isolated from conditionally targeted mice indicates PPARgamma regulates genes encoding known PPARgamma targets, additional lipid metabolism enzymes, and markers of cellular differentiation. These data reveal airway epithelial cell PPARgamma is necessary for normal lung structure and function. PMID- 16720733 TI - Anti-inflammatory effect of interleukin-10 on human neutrophil respiratory burst involves inhibition of GM-CSF-induced p47PHOX phosphorylation through a decrease in ERK1/2 activity. AB - Interleukin-10 (IL-10) exerts its anti-inflammatory properties by down-regulating polymorphonuclear neutrophil (PMN) functions such as reactive oxygen species (ROS) production via NADPH oxidase. The molecular mechanisms underlying this process are unclear. Partial phosphorylation of the NADPH oxidase cytosolic component p47(PHOX) induced by proinflammatory cytokines, such as granulocyte macrophage colony stimulating factor (GM-CSF) and tumor necrosis factor (TNF) alpha, is essential for priming ROS production by PMN. The aim of this study was to determine whether IL-10 inhibits GM-CSF- and TNFalpha-induced p47(PHOX) phosphorylation and to investigate the molecular mechanisms involved in this effect. We found that IL-10 selectively inhibited GM-CSF- but not TNFalpha induced p47PHOX phosphorylation in a concentration-dependent manner. As GM-CSF induced p47PHOX phosphorylation is mediated by extracellular signal-regulated kinase 1/2 (ERK1/2), we tested the effect of IL-10 on this pathway. We found that IL-10 inhibited GM-CSF-induced ERK1/2 activity in an immunocomplex kinase assay. This inhibitory effect was confirmed by analyzing the phosphorylation status of the endogenous substrate of ERK1/2, p90RSK, in intact PMN. Furthermore, IL-10 decreased ROS production by adherent GM-CSF-treated PMN in keeping with the higher ROS production observed in whole blood from IL-10 knockout mice compared to their wild-type counterparts. Together, these results suggest that IL-10 inhibits GM-CSF-induced priming of ROS production by inhibiting p47PHOX phosphorylation through a decrease in ERK1/2 activity. This IL-10 effect could contribute to the tight regulation of NADPH oxidase activity at the inflammatory site. PMID- 16720734 TI - Post-translational modification plays an essential role in the translocation of annexin A1 from the cytoplasm to the cell surface. AB - Annexin A1 (ANXA1) has an important role in cell-cell communication in the host defense and neuroendocrine systems. In both systems, its actions are exerted extracellularly via membrane-bound receptors on adjacent sites after translocation of the protein from the cytoplasm to the cell surface of adjacent cells. This study used molecular, microscopic, and pharmacological approaches to explore the mechanisms underlying the cellular exportation of ANXA1 in TtT/GF (pituitary folliculo-stellate) cells. LPS caused serine-phosphorylation of ANXA1 (ANXA1-S27-PO4) and translocation of the phosphorylated protein to the cell membrane. The fundamental requirement of phosphorylation for membrane translocation was confirmed by immunofluorescence microscopy on cells transfected with wild-type or mutated (S27/A) ANXA1 constructs tagged with enhanced green fluorescence protein. The trafficking of ANXA1-S27-PO4 to the cell surface was dependent on PI3-kinase and MAP-kinase. It also required HMG-coenzyme A and myristoylation. The effects of HMG-coenzyme A blockade were overcome by mevalonic acid (the product of HMG-coenzyme A) and farnesyl-pyrophosphate but not by geranyl-geranylpyrophosphate or cholesterol. Together, these results suggest that serine-27 phosphorylation is essential for the translocation of ANXA1 across the cell membrane and also identify a role for isoprenyl lipids. Such lipids could target consensus sequences in ANXA1. Alternatively, they may target other proteins in the signal transduction cascade (e.g., transporters). PMID- 16720735 TI - Aldosterone mediates angiotensin II-induced interstitial cardiac fibrosis via a Nox2-containing NADPH oxidase. AB - Angiotensin (ANG) II (AngII) and aldosterone contribute to the development of interstitial cardiac fibrosis. We investigated the potential role of a Nox2 containing NADPH oxidase in aldosterone-induced fibrosis and the involvement of this mechanism in AngII-induced effects. Nox2-/- mice were compared with matched wild-type controls (WT). In WT mice, subcutaneous (s.c.) AngII (1.1 mg/kg/day for 2 wk) significantly increased NADPH oxidase activity, interstitial fibrosis (11.5+/-1.0% vs. 7.2+/-0.7%; P<0.05), expression of fibronectin, procollagen I, and connective tissue growth factor mRNA, MMP-2 activity, and NF-kB activation. These effects were all inhibited in Nox2-/- hearts. The mineralocorticoid receptor antagonist spironolactone inhibited AngII-induced increases in NADPH oxidase activity and the increase in interstitial fibrosis. In a model of mineralocorticoid-dependent hypertension involving chronic aldosterone infusion (0.2 mg/kg/day) and a 1% Na Cl diet ("ALDO"), WT animals exhibited increased NADPH oxidase activity, pro-fibrotic gene expression, MMP-2 activity, NF-kB activation, and significant interstitial cardiac fibrosis (12.0+/-1.7% with ALDO vs. 6.3+/-0.3% without; P<0.05). These effects were inhibited in Nox2-/- ALDO mice (e.g., fibrosis 6.8+/-0.8% with ALDO vs. 5.8+/-1.0% without ALDO; P=NS). These results suggest that aldosterone-dependent activation of a Nox2-containing NADPH oxidase contributes to the profibrotic effect of AngII in the heart as well as the fibrosis seen in mineralocorticoid-dependent hypertension. PMID- 16720736 TI - Oxidative stabilization of iso-1-cytochrome c by redox-inspired protein engineering. AB - Iso-1-cytochrome c, as any other hemeprotein, is able to react with hydrogen peroxide and to engage in the peroxidase cycle. However, peroxidases are irreversibly inactivated by their substrate, hydrogen peroxide. The oxidative inactivation of hemeproteins is mechanism based and arises as the consequence of unproductive electron abstraction reactions. Protein elements, such as the porphyrin ring or the protein backbone, act as simultaneous and competing electron sources even in the presence of exogenous reducing substrates, leading to a decline in activity. It is hypothetically possible to alter the intramolecular electron transfer pathways by direct replacement of low redox potential residues around the active site; as a consequence, the inactivation process would be delayed or even suppressed. To demonstrate this hypothesis, a redox-inspired strategy was implemented until an iso-1-cytochrome c variant fully stable at catalytic concentrations of hydrogen peroxide was obtained. This variant, harboring the N52I,W59F,Y67F,K79A,F82G substitutions, preserved the catalytic performance of the parental protein but achieved a 15-fold higher total turnover number. The phenotype of this variant was reflected in the stability of its electronic components, allowing identification of a protein-based radical intermediate mechanistically similar to Compound I of classical peroxidases. The results presented here clearly demonstrate that redox-inspired protein engineering is a useful tool for the rational modulation of intramolecular electron transfer networks. PMID- 16720737 TI - Positive effects of 17beta-estradiol on insulin sensitivity in aged ovariectomized female rats. AB - Aging is associated with insulin resistance, which represents a common factor in age-related diseases. We aimed to determine the role of 17beta-estradiol on insulin sensitivity and memory during aging using ovariectomized rats (2-26 months of age) treated with physiological doses of 17beta-estradiol. Our results indicate a lack of effect of 17beta-estradiol replacement on spatial memory assessed in a water maze. Conversely, estradiol treatment improved insulin sensitivity in aging rats. These data imply that relatively low doses of 17beta estradiol may have beneficial effects on glucose homeostasis due to the protective effects of estrogen. However, estradiol treatment used in the present study did not prevent memory impairment associated with aging. PMID- 16720738 TI - Age-related loss of the DNA repair response following exposure to oxidative stress. AB - Young (4- to 6-month-old) and aged (24- to 28-month-old) mice were exposed to 2 nitropropane (2-NP), a DNA oxidizing agent, and the ability to induce DNA polymerase beta (beta-pol) and AP endonuclease (APE) was determined. In contrast to the inducibility of these gene products in response to oxidative damage in young mice, aged mice showed a lack of inducibility of beta-pol and APE. APE protein level and endonuclease activity were both reduced 40% (p<.01) in response to 2-NP. Accordingly, the accumulation of DNA repair intermediates in response to 2-NP differed with age. Young animals accumulated 3'OH-containing DNA strand breaks, whereas the aged animals did not. A role for p53 in the difference in DNA damage response with age is suggested by the observation that the accumulation of p53 protein in response to DNA damage in young animals was absent in the aged animals. Our results are consistent with a reduced ability to process DNA damage with age. PMID- 16720739 TI - The effect of aging on the chaperone concentrations in the hepatic, endoplasmic reticulum of male rats: the possible role of protein misfolding due to the loss of chaperones in the decline in physiological function seen with age. AB - The endoplasmic reticulum (ER) chaperones are highly conserved proteins that catalyze the posttranslational processing of all secretory and membrane proteins. Our studies suggest that chaperone declines are one of the two central defects in Alzheimer's disease. We propose that similar declines in other organ systems underlie the physiological deficits of aging. Rats were maintained in a colony from age 21 days to death. Animals were killed at regular intervals, and hepatic, ER chaperone contents were determined by immunoblotting. ERp55, ERp57, ERp72, BiP, and calnexin constitutive levels declined 30%-50% with age. Calreticulin was unaffected. BiP (also known as GRP78), ERp55, and ERp57 showed marked swings with peaks occurring in midwinter and midsummer. This cyclics declined 73% with age. Considering the role of the ER chaperones in membrane and secretory protein posttranslational processing, these data support the concept that their loss could lead to many of the physiological declines associated with aging. PMID- 16720740 TI - daf-16 protects the nematode Caenorhabditis elegans during food deprivation. AB - Inhibition of either the insulin-like or target of rapamycin (TOR) pathways in the nematode Caenorhabditis elegans extends life span. Here, we demonstrate that starvation and inhibition of the C. elegans insulin receptor homolog (daf-2) elicits a daf-16-dependent up-regulation of a mitochondrial superoxide dismutase (sod-3). We also find that although heat and oxidative stress result in nuclear localization of the DAF-16 protein, these stressors do not activate a SOD-3 reporter, suggesting that nuclear localization alone may not be sufficient for transcriptional activation of DAF-16. We show that inhibition of either TOR activity or key components of the cognate translational machinery (eIF-4G and EIF 2B homologs) increases life span by both daf-16-dependent and -independent mechanisms. Finally, we demonstrate that at least one nematode hexokinase is localized to the mitochondria. We propose that the increased life spans conferred by alterations in both the TOR and insulin-like pathways function by inappropriately activating food-deprivation pathways. PMID- 16720741 TI - Nuclear architecture and disease. PMID- 16720743 TI - Randomized controlled trials involving multidisciplinary interventions in the community. PMID- 16720742 TI - The endeavor of high maintenance homeostasis: resting metabolic rate and the legacy of longevity. AB - Metabolism, the continuous conversion between structural molecules and energy, is life in essence. Size, metabolic rate, and maximum life span appear to be inextricably interconnected in all biological organisms and almost follow a "universal" law. The notion of metabolic rate as the natural "rate of living" filled most of the academic discussion on aging in the early 20th century to be later replaced by the free-radical theory of aging. We argue that the rate of living theory was discarded too quickly and that studying factors affecting resting metabolic rate during the aging process may provide great insight into the core mechanisms explaining differential longevity between individuals, and possibly the process leading to frailty. We predict that measures of resting metabolic rate will be introduced in geriatric clinical practice to gather information on the degree of multisystem dysregulation, exhaustion of energy reserve, and risk of irreversible frailty. PMID- 16720744 TI - Anemia in old age is associated with increased mortality and hospitalization. AB - BACKGROUND: Anemia is common in old age and has been shown to affect older persons' physical function. To more fully understand the detrimental health effects of anemia, we examined the relationship of anemia with death and hospitalization outcomes in a large community-based sample of older persons. METHODS: Data are from 3607 persons, aged 71 years or older, participating in the National Institute on Aging (NIA)-sponsored Established Populations for Epidemiologic Studies of the Elderly (EPESE) study. Anemia was defined according to World Health Organization (WHO) criteria as a hemoglobin concentration below 12 g/dL in women and below 13 g/dL in men. Data on subsequent mortality and hospital admissions over 4 years were obtained from death records and the Medicare database. RESULTS: Anemia was present in 451 of the 3607 (12.5%) participants. During the follow-up period, anemic persons were more likely to die than were nonanemic persons (37.0% vs 22.1%, p<.001). Also, anemic persons were more often hospitalized (65.9% vs 54.6%, p<.001) and spent more days in hospital (25.0 vs 13.7, p<.001). After adjustment for demographics and baseline comorbidities, anemia significantly predicted subsequent mortality and hospitalization (relative risk=1.61, 95% confidence interval, 1.34-1.93; and relative risk=1.27, 95% confidence interval, 1.12-1.45, respectively). After excluding persons with prevalent diseases at baseline, anemia remained significantly associated with increased risks of mortality and hospitalization. A higher hemoglobin level was significantly associated with lower risks of mortality and hospitalization (p for trend<.001 for both). CONCLUSIONS: These findings indicate that late-life anemia characterizes persons at risk for important clinical health outcomes, and demonstrate the importance of clinical awareness of anemia even if the person is without apparent clinical disease. PMID- 16720745 TI - Effects of aerobic and resistive exercise training on glucose disposal and skeletal muscle metabolism in older men. AB - BACKGROUND: Aging is associated with insulin resistance, primarily as a result of physical inactivity and increased abdominal obesity. We hypothesized that aerobic (AEX) or resistive (RT) exercise training would result in comparable improvements in glucose disposal in older men, but that there would be different metabolic adaptations in skeletal muscle. METHODS: Thirty-nine older (63+/-1 years, mean+/ standard error of the mean), overweight and obese (body mass index=30.3+/-0.4 kg/m2) men were assigned to AEX (treadmill walking and/or jogging, n=19) or RT (upper and lower body, n=20) programs 3 d/wk for 6 months, with 9 completing AEX and 13 completing RT. Testing before and after the exercise programs included body composition, euglycemic-hyperinsulinemic clamps, and vastus lateralis muscle biopsies. RESULTS: Maximal oxygen consumption (VO2max) increased by 16% after AEX (p<.01), while leg and arm muscle strength increased by 45+/-5% and 27+/-5% after RT (p<.0001). Although participants were monitored to maintain their body weight during the exercise program, body weight decreased by 2% after AEX (p<.05), and increased by 2% after RT (p<.05). Whole-body glucose disposal, determined during the last 30 minutes of a 2-hour 480 pmol/m2/min euglycemic-hyperinsulinemic clamp, increased comparably by 20%-25% after AEX (51+/-5 to 61+/-5 microM/kgfat free mass/min, p<.05) and RT (49+/-3 to 58+/-3 microM/kgfat-free mass/min, p<.05). The increase in vastus lateralis muscle glycogen synthase fractional activity in response to insulin stimulation was significantly higher after AEX compared to after RT (279+/-59% compared to 100+/-28% change, p<.05). Neither AEX nor RT altered muscle glycogen synthase total activity, glycogen content, or levels of phosphotidylinositol 3-kinase. CONCLUSION: These results suggest that AEX and RT result in comparable improvements in glucose metabolism in older men, whereas an increase in insulin activation of glycogen synthase occurred only with AEX. These improvements in insulin sensitivity could reduce the risk of metabolic syndrome and type 2 diabetes and attenuate the development of cardiovascular disease. PMID- 16720746 TI - Depressive symptoms and changes in body weight exert independent and site specific effects on bone in postmenopausal women exercising for 1 year. AB - BACKGROUND: Lower bone mineral density (BMD) has been documented in clinically depressed populations, and depression is the second most common chronic medical condition in general medical practice. Therefore, the purpose of this study was to determine whether depressive symptoms, vitality, and body weight changes were related to 1-year BMD changes after accounting for covariates. METHODS: Healthy postmenopausal women (n=320; 40-65 years) were recruited, and 266 women completed the study. Participants were 3-10 years postmenopausal, sedentary, and either taking hormone replacement therapy (1-3.9 years) or not taking it (at least 1 year). Exclusion criteria were: current smoking status, history of fractures, low BMD, body mass index>32.9 or <19.0, or use of bone altering medications. Regional BMD was measured from dual-energy x-ray absorptiometry at baseline and 1 year. Self-reported depressive symptoms and vitality were measured using standard questionnaires. RESULTS: Both the vitality and depressive symptoms scores were related to BMD changes at the femur neck but not at the greater trochanter or spine. Weight change was a predictor of BMD changes in the trochanter and spine but not in the femoral neck. Weight change and vitality and/or depressive symptoms had differential and site-specific effects on BMD changes at the hip. Vitality and depressive symptoms related to femoral neck changes and weight change related to greater trochanter changes. CONCLUSIONS: The negative impact of depressive symptoms on BMD in this population of postmenopausal women was independent of body weight or other behavioral factors such as calcium compliance or exercise. PMID- 16720747 TI - Gender differences in functioning after hip fracture. AB - BACKGROUND: Hip fracture is a significant health problem for men and women; between 25 and 30 percent of all hip fractures are sustained by men. Relatively little is known about gender differences in functional outcomes after hip fracture. The purpose of the current study is to compare post-hip fracture functional recovery of men and women. METHODS: A sample of 674 patients age 65 or older were recruited as part of the Baltimore Hip Studies and were followed longitudinally for 1 year following fracture. Information on prefracture status and hospital course of treatment was collected as well as functional data at baseline, 2, 6, and 12 months postfracture. Data were analyzed longitudinally using Generalized Estimating Equations (GEEs). RESULTS: Men in the study were generally younger and suffered greater comorbidity at time of fracture. Men further suffered higher mortality in the year following fracture. Among survivors, little difference between men and women was seen in patterns of recovery of function following fracture. CONCLUSIONS: Hip fracture is not a problem affecting just women. Recovery following fracture for men is probably no better than that for women, even after mortality differentially eliminates the frailest male participants. However, psychosocial factors, greater comorbidity, and higher rates of certain complications among men may require adjustments to interventions designed to restore function. Further research into the consequences of hip fracture for men and women is needed. PMID- 16720748 TI - Age-related differences in the effect of a perceived threat to stability on postural control. AB - BACKGROUND: Studies indicate that the strategy for postural control may be affected by psychological factors, and that young adults respond to perceived threat to stability by increasing their ankle stiffness. The objective of this study was to compare the postural control strategy adopted by young and old adults when faced with postural threat induced by manipulating surface height. METHODS: Sixty elderly (mean age 77.5+/-4.4 years) and 20 young volunteers (mean age 21.5+/-3.7 years) participated in the study. Movement of the center of pressure (COP) was recorded for 60 seconds with a portable force-plate, as participants stood with eyes open and closed, both at ground level and 85 cm above ground level. Analysis of variance and Tukey-Kramer tests were conducted to determine the effects of age, postural threat, and vision on mean power frequency (MPF), on amplitude variability both in the anterior-posterior (AP) and medio lateral (ML) directions, and on mean COP sway velocity, with the significance level set at p=.05. RESULTS: Postural threat did not affect COP measures for the young adults, whereas it induced greater changes in MPF in the AP and ML directions in the elderly participants. A similar trend was observed in the elderly group for amplitude variability in the ML direction and for mean COP sway velocity. CONCLUSIONS: In comparison with young adults, elderly adults tend to have an exaggerated postural response to conditions that are perceived as threatening to stability. This response involves primarily increases in MPF, with limited compensatory decreases in amplitude variability observed only in the ML direction. PMID- 16720749 TI - Metabolic syndrome, diabetes, and cardiovascular disease in an elderly Caucasian cohort: the Italian Longitudinal Study on Aging. AB - BACKGROUND: The metabolic syndrome (MetS) is represented by a cluster of risk factors for cardiovascular diseases (CVDs). In spite of its high frequency and strong association with morbidity and mortality in the adult population, little is known about its magnitude in elderly persons. METHODS: We assessed the prevalence of MetS by diabetic status and sex in the participants in the Italian Longitudinal Study on Aging (ILSA), a population-based study on a sample of 5632 individuals 65-84 years old at baseline (1992). We measured the association of MetS with stroke, coronary heart disease, and diabetes at baseline and with CVD mortality at 4-year follow-up. RESULTS: The prevalence of MetS was 25.9% in nondiabetic men and 55.2% in nondiabetic women; in diabetic individuals it was 64.9% and 87.1% in men and women, respectively. At baseline, in both men and women there was a significant association with stroke (odds ratio [OR]=1.67, 95% confidence interval [CI], 1.02-2.75 in men and OR=1.72, CI, 1.01-2.93 in women) and diabetes (OR=4.58, CI, 3.12-6.74 in men and OR=5.15, CI, 3.23-8.20 in women). A significant association with chronic heart disease was found in men only (OR=1.40; CI, 1.02-1.97). During the approximately 4-year follow-up, nondiabetic men with MetS had a risk of CVD mortality 12% higher compared to those without MetS, whereas no significant differences were found in women. CONCLUSIONS: MetS is very common in aged Italians, and it is associated with stroke and diabetes in both sexes, and with chronic heart disease in men. In men, it increases significantly the risk of CVD mortality. PMID- 16720750 TI - Incidence and predictors of all and preventable adverse drug reactions in frail elderly persons after hospital stay. AB - BACKGROUND: Adverse drug reactions (ADR) negatively impact life quality and are sometimes fatal. This study examines the incidence and predictors of all and preventable ADRs in frail elderly persons after hospital discharge, a highly vulnerable but rarely studied population. METHODS: The design was a prospective cohort study involving 808 frail elderly persons who were discharged from 11 Veteran Affairs hospitals to outpatient care. The main outcome measure was number of ADRs per patient as determined by blinded geriatrician and geropharmacist pairs using Naranjo's ADR algorithm. For all ADRs (possible, probable, or definite), preventability was assessed. Discordances were resolved by consensus conferences. RESULTS: Overall, 33% of patients had one or more ADRs for a rate of 1.92 per 1000 person-days of follow-up. The rate for preventable ADRs was 0.71 per 1000 person-days of follow-up. Independent risk factors for all ADRs were number of medications (adjusted [Adj.] hazard ratio [HR], 1.07; 95% confidence interval [CI], 1.05-1.10 per medication), use of warfarin (Adj. HR, 1.51; 95% CI, 1.22-1.87), and (marginally) the use of benzodiazepines (Adj. HR, 1.23; 95% CI, 0.95-1.58). Counterintuitively, use of sedatives and/or hypnotics was inversely related to ADR risk (Adj. HR, 0.14; 95% CI, 0.04-0.57). Similar trends were seen for number of medications and warfarin use as predictors of preventable ADRs. CONCLUSIONS: ADRs are very common in frail elderly persons after hospital stay, and polypharmacy and warfarin use consistently increase the risk of ADRs. PMID- 16720751 TI - Outcome of Alzheimer's disease: potential impact of cholinesterase inhibitors. AB - BACKGROUND: Alzheimer's disease is fast becoming a major public health concern with serious economic consequences. The cholinesterase inhibitors (CEIs) offer some benefit in the symptomatic treatment of the disease. This study aims to investigate the effect of CEIs on three clinically relevant domains (rapid cognitive decline, institutionalization, and weight loss) in patients with Alzheimer's disease. METHODS: A prospective observational study was performed in which a population of 455 Alzheimer's disease patients were recruited and followed up for at least 1 year between 1994 and 2002. Patients were reevaluated at 6 monthly intervals using standardized neurocognitive and geriatric evaluations in addition to complete clinical examination, standard paraclinical investigations, and recording of treatment received. RESULTS: The risk of rapid cognitive deterioration was significantly decreased in patients taking CEIs for at least 1 year compared to untreated patients (odds ratio [OR]=0.56, 95% confidence interval [CI], 0.34-0.93; p=.025). The potential benefit of CEI use was also found on institutionalization (OR=0.2, 95% CI, 0.08-0.48; p<.001) and weight loss (OR=0.56, 95% CI, 0.32-0.97; p=.039) after 1 year of follow-up. CONCLUSION: The special interest of this study is that all patients were recruited and followed in the same center with the same management care plan and the same medical team. This follow-up offers us a unique opportunity to compare the 1-year evolution of the disease in clinical practice before and after the marketing of CEIs and allows us to demonstrate a clinically significant improvement in patient outcome over time. PMID- 16720752 TI - Does testosterone mediate cognitive decline in elderly men? PMID- 16720753 TI - Ritonavir, saquinavir, and efavirenz, but not nevirapine, inhibit bile acid transport in human and rat hepatocytes. AB - Human immunodeficiency virus-infected patients on antiretroviral drug therapy frequently experience hepatotoxicity, the underlying mechanism of which is poorly understood. Hepatotoxicity from other compounds such as bosentan and troglitazone has been attributed, in part, to inhibition of hepatocyte bile acid excretion. This work tested the hypothesis that antiretroviral drugs modulate hepatic bile acid transport. Ritonavir (28 microM), saquinavir (15 microM), and efavirenz (32 microM) inhibited [(3)H]taurocholate transport in bile salt export pump expressing Sf9-derived membrane vesicles by 90, 71, and 33%, respectively. In sandwich-cultured human hepatocytes, the biliary excretion index (BEI) of [(3)H]taurocholate was maximally decreased 59% by ritonavir, 39% by saquinavir, and 20% by efavirenz. Likewise, in sandwich-cultured rat hepatocytes, the BEI of [(3)H]taurocholate was decreased 100% by ritonavir and 94% by saquinavir. Sodium dependent and -independent initial uptake rates of [(3)H]taurocholate in suspended rat hepatocytes were significantly decreased by ritonavir, saquinavir, and efavirenz. [(3)H]Taurocholate transport by recombinant NTCP and Ntcp was inhibited by ritonavir (IC(50) = 2.1 and 6.4 microM in human and rat, respectively), saquinavir (IC(50) = 6.7 and 20 microM, respectively), and efavirenz (IC(50) = 43 and 97 microM, respectively). Nevirapine (75 microM) had no effect on bile acid transport in any model system. In conclusion, ritonavir, saquinavir, and efavirenz, but not nevirapine, inhibited both the hepatic uptake and biliary excretion of taurocholate. PMID- 16720754 TI - Ethanol destabilizes liver Gal beta l, 4GlcNAc alpha2,6-sialyltransferase, mRNA by depleting a 3'-untranslated region-specific binding protein. AB - Asialoconjugates are viable biomarkers for alcohol abuse. We previously showed that chronic ethanol feeding down-regulated liver Gal beta l, 4GlcNAc alpha2,6 sialyltransferase (ST6Gal l) mRNA by destabilizing it. Since RNA-binding proteins are known to stabilize many eukaryotic mRNAs by interacting with the 3' untranslated region (UTR), we have delineated the possible mechanism by which ethanol destabilizes ST6Gal l mRNA. Using (32)P-labeled RNA probes generated from a 2.7-kb 3'-UTR of ST6Gal l mRNA, we identified a liver cytosolic 41-kDa specific binding protein that interacts with its 3'-UTR domain and protects it from degradation in normal rat liver but disappears after chronic ethanol treatment. Mapping of the binding region revealed that four RNA probes of 80-base pair (bp) length spanning the 304 bp of the 3'-UTR of ST6Gal l mRNA showed equal binding intensity. The corresponding cDNA sequences for the four 80-bp RNA probes share the 13-bp consensus sequence. Mutagenesis analysis identified that four nucleotides, AG and TC, among the consensus sequences were critical for the RNA protein interaction. Therefore, 5'-CAGCCTCCTCCCT-3' serves as a cis-element critically involved in this interaction. The RNA-protein complex formation progressively decreased with increasing dietary ethanol, resulting in its virtual disappearance with 36% of the dietary calories as ethanol. Concomitantly, the same ethanol diet decreased sialic acid index of plasma apolipoprotein J by 45% (p < 0.05). Thus, depletion of a binding protein that specifically interacts with its 3'-UTR region of ST6Gal l mRNA may account for its destabilization and consequent appearance of asialoconjugates as alcohol biomarkers. PMID- 16720755 TI - Biotransformation of glyceryl trinitrate by rat hepatic microsomal glutathione S transferase 1. AB - Although the biotransformation of organic nitrates by the cytosolic glutathione S transferases (GSTs) is well known, the relative contribution of the microsomal GST (MGST1) to nitrate biotransformation has not been described. We therefore compared the denitration of glyceryl trinitrate (GTN) by purified rat liver MGST1 and cytosolic GSTs. Both MGST1 and cytosolic GSTs catalyzed the denitration of GTN, but the activity of MGST1 toward GTN was 2- to 3-fold higher. To mimic oxidative/nitrosative stress in vitro, we treated enzyme preparations with hydrogen peroxide, S-nitrosoglutathione, and peroxynitrite. Both oxidants and nitrating reagents increased the activity of MGST1 toward the GST substrate, 1 chloro-2,4-dinitrobenzene (CDNB) whereas these treatments inhibited GTN denitration by MGST1. Alkylation of the sole cysteine residue of MGST1 by N ethylmaleimide markedly increased enzyme activity with CDNB as substrate but decreased the rate of GTN denitration. In aortic microsomes from GTN-tolerant animals, there was a decreased abundance of MGST1 dimers and trimers. In hepatic microsomes from GTN-tolerant animals, GTN biotransformation was unaltered whereas the rate of CDNB conjugation was doubled, suggesting that chronic GTN exposure causes structural modifications to the enzyme, resulting in increased activity to certain substrates. Collectively, these data indicate that MGST1 contributes significantly to the biotransformation of GTN and that chemical modification of the microsomal enzyme has differential effects on the catalytic activity toward different substrates. PMID- 16720756 TI - Evaluation of original dual thromboxane A2 modulators as antiangiogenic agents. AB - Angiogenesis is a promising target for the therapy of several diseases including cancer. This study was undertaken to characterize the antiangiogenic properties of a series of original dual thromboxane A(2) (TXA(2)) inhibitors derived from torasemide, a marketed loop diuretic with TXA(2) antagonistic properties, by evaluating their effects on human endothelial cell migration, adhesion, and viability in vitro, as well as in the ex vivo rat aortic ring assay. All drugs tested exhibited a marked affinity toward human platelet TXA(2) receptor, significantly prevented platelet aggregation induced by U-46,619, a stable TXA(2) receptor agonist, and inhibited platelet TXA(2) synthase without affecting cyclooxygenase (COX)-1 or COX-2 enzymatic activities. These dual TXA(2) inhibitors dose dependently inhibited endothelial cell migration in chemotaxis assays using vascular endothelial growth factor (VEGF) as a chemoattractant but failed to affect cell adhesion and viability. The highest rates of cell migration inhibition were obtained with original compounds BM-567 and BM-573 (50.3 and 59.4% inhibition, respectively) when used at the final concentration of 10 microM. In addition, pretreatment of endothelial cells with these two drugs significantly prevented U-46,619-induced intracellular Ca(2+) pool mobilization, thus suggesting a mechanistic link between inhibition of the TXA(2) pathway and reduced endothelial cell migration. Treatment of rat aortic explants with U 46,619 (9,11-dideoxy-9,11-methanoepoxy-prostaglandin F(2)) significantly enhanced vessel sprouting whereas aortic rings treated with some of the compounds, including BM-567 (N-n-pentyl-N'-[2-(cyclohexylamino)-5-nitrobenzenesulfonyl]urea) and BM-573 (N-tert-butyl-N'-[5-nitro-2-p-toluylaminobenzenesulfonyl]urea), showed a significant decrease in vessel sprouting, which was not reversed by the addition of VEGF. These data suggest that our original dual TXA(2) inhibitors bear antiangiogenic properties, mainly by inhibiting endothelial cell migration. PMID- 16720757 TI - alpha4beta2 nicotinic receptors with high and low acetylcholine sensitivity: pharmacology, stoichiometry, and sensitivity to long-term exposure to nicotine. AB - alpha4 and beta2 nicotinic acetylcholine receptor (nAChR) subunits expressed heterologously assemble into receptors with high (HS) and low (LS) sensitivity to acetylcholine (ACh); their relative proportions depend on the alpha4to beta2 ratio. In this study, injection of oocytes with 1:10 alpha4/beta2 subunit cDNA ratios favored expression of HS alpha4beta2 nAChRs, as evidenced by monophasic ACh concentration-response curves, whereas injections with 10:1 cDNA ratios favored expression of LS alpha4beta2 receptors. The stoichiometry was inferred from the shifts in the ACh EC(50) values caused by Leu to Thr mutations at position 9' of the second transmembrane domain of alpha4 and beta2. The 1:10 injection ratio produced the (alpha4)(2)(beta2)(3) stoichiometry, whereas 10:1 injections produced the (alpha4)(3)(beta2)(2) stoichiometry. The agonists epibatidine, 3-[2(S)-azetidinylmethoxy]pyridine (A-85380), 5-ethoxy-metanicotine (TC-2559), cytisine, and 3-Br-cytisine and the antagonists dihydro-beta erythroidine and d-tubocurarine were more potent at HS receptors. TC-2559 was more efficacious than ACh at HS receptors but was a partial agonist at LS receptors. Epibatidine was more efficacious than ACh at LS receptors and a partial agonist at HS receptors. Cytisine and 5-halogenated cytisines had moderate efficacy at LS receptors but had almost no efficacy at HS receptors. By exploiting the differential effects of ACh, TC-2559 and 5-I-cytisine we evaluated the effects of long-term exposure to nicotine on HS and LS receptors expressed in Xenopus laevis oocytes after cDNA injections or microtransplantation of alpha4beta2 receptors assembled in human embryonic kidney 293 cells. We conclude that nicotine up-regulates HS alpha4beta2 receptors, probably by influencing the assembly of receptors rather than by altering the functional state of LS alpha4beta2 nAChRs. PMID- 16720758 TI - The role of growth factors and related agents in accelerating fracture healing. PMID- 16720759 TI - Dislocation of the knee. PMID- 16720760 TI - The effect of a neck-retaining femoral implant on leg-length inequality following total hip arthroplasty: a radiological study. AB - The aim of this study was to assess whether a femoral component which retained the neck reduced the incidence of leg-length inequality following total hip arthroplasty. A retrospective review was undertaken of 130 consecutive primary total hip arthroplasties performed between April 1996 and April 2004 using such an implant. There were 102 suitable patients for the study. Standardised pre- and post-operative pelvic radiographs were measured by an independent investigator to the nearest millimetre. The leg-length inequality was reduced from a mean pre operative value of -0.71 cm to a mean of 0.11 cm post-operatively. Of the 102 patients 24 (23.5%) had an equal leg-length post-operatively, and 95 (93.1%) had a leg-length inequality between -1 cm and 1 cm. PMID- 16720761 TI - Continuous monitoring of the performance of hip prostheses. AB - New brands of joint prosthesis are released for general implantation with limited evidence of their long-term performance in patients. The CUSUM continuous monitoring method is a statistical testing procedure which could be used to provide prospective evaluation of brands as soon as implantation in patients begins and give early warning of poor performance. We describe the CUSUM and illustrate the potential value of this monitoring tool by applying it retrospectively to the 3M Capital Hip experience. The results show that if the clinical data and methodology had been available, the CUSUM would have given an alert to the underperformance of this prosthesis almost four years before the issue of a Hazard Notice by the Medical Devices Agency. This indicates that the CUSUM can be a valuable tool in monitoring joint prostheses, subject to timely and complete collection of data. Regional or national joint registries provide an opportunity for future centralised, continuous monitoring of all hip and knee prostheses using these techniques. PMID- 16720762 TI - Biomechanical reconstruction of the hip: a randomised study comparing total hip resurfacing and total hip arthroplasty. AB - We have compared the biomechanical nature of the reconstruction of the hip in conventional total hip arthroplasty (THA) and surface replacement arthroplasty (SRA) in a randomised study involving 120 patients undergoing unilateral primary hip replacement. The contralateral hip was used as a control. Post-operatively, the femoral offset was significantly increased with THA (mean 5.1 mm; -2.8 to 11.6) and decreased with SRA (mean -3.3 mm; -8.9 to 8.2). Femoral offset was restored within sd 4 mm in 14 (25%) of those with THA and in 28 (57%) of the patients receiving SRA (p < 0.001). In the THA group, the leg was lengthened by a mean of 2.6 mm (-6.04 to +12.9), whereas it was shortened by a mean of 1.9 mm ( 7.1 to +2.05) in the SRA group, compared with the contralateral side. Leg-length inequality was restored within sd 4 mm in 42 (86%) of the SRA and 33 (60%) of the THA patients. The radiological parameters of acetabular reconstruction were similar in both groups. Restoration of the normal proximal femoral anatomy was more precise with SRA. The enhanced stability afforded by the use of a large diameter femoral head avoided over-lengthening of the limb or increased offset to improve soft-tissue tension as occurs sometimes in THA. In a subgroup of patients with significant pre-operative deformity, restoration of the normal hip anatomy with lower pre-operative femoral offset or significant shortening of the leg was still possible with SRA. PMID- 16720763 TI - The relationship between acetabular retroversion and osteoarthritis of the hip. AB - Excessive acetabular cover secondary to a retroverted acetabulum causes pincer impingement, which may cause early osteoarthritis of the hip. Our aim was to determine if there was a relationship between acetabular version and osteoarthritis of the hip. Using image processing and analysis software we studied 117 CT images of the hip in patients aged less than 65 years who had undergone a CT virtual colonoscopy. The mean CT joint space of the 18 hips with acetabular retroversion was narrower compared with the 99 hips with normal acetabular alignment (p < 0.0001). A correlation of r = 0.46 (p < 0.01) was found between right hip acetabular version and the mean right hip joint space and of r = 0.31 (p = 0.02) between left hip acetabular version and the mean left hip joint space. Acetabular retroversion is associated with radiological evidence of osteoarthritis of the hip. An understanding of the mechanical basis of osteoarthritis of the hip allows early treatment of the underlying structural abnormality and prevents progression of the degenerative condition. PMID- 16720764 TI - In-cement technique for revision hip arthroplasty. AB - The in-cement technique for revision hip arthroplasty involves retaining the original cement-bone interface. This has been proven to be a biomechanically stronger method than recementing after complete removal of the original cement mantle. This study reviewed a series of 54 consecutive revision hip arthroplasty procedures, using the in-cement technique, between November 1999 and November 2003. Clinical and radiological follow-up included functional assessment. There were 54 procedures performed in 51 patients, whose mean age at surgery was 70.3 years (45 to 85). A total of 42 were available at a mean follow-up of 29.2 months (6 to 51). There was no radiological evidence of loosening. Functional assessments were available for 40 patients who had a mean Harris hip score of 85.2 (51.9 to 98.5), a mean Oxford hip score of 19.6 (12 to 41), a mean UCLA activity profile score of 5.9 (3 to 8) and a mean SF-36 score of 78.0 (31.6 to 100). The in-cement technique provides consistent, high functional outcomes and should be considered in appropriately selected cases. PMID- 16720765 TI - Patellar resurfacing in total knee replacement: a ten-year randomised prospective trial. AB - A series of 100 consecutive osteoarthritic patients was randomised to undergo total knee replacement using a Miller-Galante II prosthesis, with or without a cemented polyethylene patellar component. Knee function was evaluated using the American Knee Society score, Western Ontario and McMaster University Osteoarthritis index, specific patellofemoral-related questions and radiographic evaluation until the fourth post-operative year, then via questionnaire until ten years post-operatively. A ten-point difference in the American Knee Society score between the two groups was considered a significant change in knee performance, with alpha and beta levels of 0.05. The mean age of the patients in the resurfaced group was 71 years (53 to 88) and in the non-resurfaced group was 73 years (54 to 86). After ten years 22 patients had died, seven were suffering from dementia, three declined further participation and ten were lost to follow-up. Two patients in the non-resurfaced group subsequently had their patellae resurfaced. In the resurfaced group one patient had an arthroscopic lateral release. There was no significant difference between the two treatment groups: both had a similar deterioration of scores with time, and no further patellofemoral complications were observed in either group. We are unable to recommend routine patellar resurfacing in osteoarthritic patients undergoing total knee replacement on the basis of our findings. PMID- 16720767 TI - Results of arthrodesis in neuropathic feet. AB - We describe the results of arthrodesis for the treatment of recurrent acute neuropathic bone disease in 24 feet and of chronic disease with deformity in 91 feet, undertaken between January 1984 and December 2003. All were due to leprosy. Correction of the deformity was achieved in 80 of 106 feet (76%) and fusion in 97 of 110 feet (88%). In the 24 feet in which recurrent neuropathic bone disease was the reason for surgery, 17 (71%) obtained stability while in seven (29%) symptoms recurred postoperatively. Complications were experienced following 58 of the 110 operations (53%). In patients presenting primarily with deformity with a minimum follow-up of two years (79 feet), there was a reduced frequency of ulceration in 40 (51%). Normal footwear could be worn by 32 patients (40%) after surgery, while 40 (51%) required a moulded insole. Arthrodesis of the ankle in the neuropathic foot due to leprosy has a good overall rate of success although the rate of complications is high. PMID- 16720766 TI - Percutaneous drilling for the treatment of secondary osteonecrosis of the knee. AB - Osteonecrosis of the knee comprises two separate disorders, primary spontaneous osteonecrosis which is often a self-limiting condition and secondary osteonecrosis which is associated with risk factors and a poor prognosis. In a series of 61 knees (38 patients) we analysed secondary osteonecrosis of the knee treated by a new technique using multiple small percutaneous 3 mm drillings. Total knee replacement was avoided in 59 knees (97%) at a mean follow-up of 3 years (2 to 4). Of the 61 knees, 56 (92%) had a successful clinical outcome, defined as a Knee Society score greater than 80 points. The procedure was successful in all 24 knees with small lesions compared with 32 of 37 knees (86%) with large lesions. All the procedures were performed as day cases and there were no complications. This technique appears to have a low morbidity, relieves symptoms and delays more invasive surgery. PMID- 16720768 TI - Single-stage bilateral total shoulder arthroplasty: a preliminary study. AB - We compared six patients with a mean age of 70 years (49 to 80) with severe bilateral, painful glenohumeral joint destruction who underwent a single-stage bilateral total shoulder replacement, with eight patients of mean age 61 years (22 to 89) who underwent bilateral total shoulder replacement in two stages, at a mean interval of 18 months (6 to 43). The overall function, pain and strength improved significantly in both groups. The subjective shoulder value, relative Constant score, active external rotation and the strength were improved significantly more in the single-stage group. Active elevation, abduction and overall function improved, significantly more in the single-stage group. Both the total duration of hospitalisation and the time off work per shoulder were substantially shorter in the single-stage group. The overall rate of complication was lower in the single-stage group. Our findings indicated that single-stage bilateral total shoulder replacement yielded significantly better clinical results with shorter hospitalisation and rehabilitation than staged replacement, and was not associated with any increase in complications. PMID- 16720769 TI - The effects of operative delay on the relief of neuropathic pain after injury to the brachial plexus: a review of 148 cases. AB - We investigated the effect of delay before nerve repair on neuropathic pain after injury to the brachial plexus. We studied 148 patients, 85 prospectively and 63 retrospectively. The mean number of avulsed spinal nerves was 3.2 (1 to 5). Pain was measured by a linear visual analogue scale and by the peripheral nerve injury scale. Early repair was more effective than delayed repair in the relief from pain and there was a strong correlation between functional recovery and relief from pain. PMID- 16720770 TI - Comminuted fractures of the radial head treated by the Judet floating radial head prosthesis. AB - Excision is not a suitable treatment for all comminuted fractures of the radial head. In elbows where instability can be predicted, a replacement arthroplasty of the radial head is more effective. The aim of this paper was to present the medium-term results of the Judet floating radial head prosthesis. This operation was performed on 14 patients between 1992 and 2003, of whom 12 were reviewed at a mean follow-up of five years and three months (1 to 12 years). The outcome was assessed using the Mayo elbow performance score and a modified Disability of Arm Shoulder Hand (DASH) questionnaire. There were six excellent results, four good, one fair and one poor, as graded by the Mayo score. The mean DASH score was 23.9/100 (0 to 65.8/100). The only significant complication occurred in one patient who developed a severe complex regional pain syndrome. There were no patients with secondary instability of the elbow, implant loosening, cubitus valgus, osteoporosis of the capitellum, or pain in the forearm and wrist. Our experience, combined with that of other authors using this device, has encouraged us to continue using the Judet prosthesis in comminuted fractures of the elbow where instability is a potential problem. PMID- 16720771 TI - Posterior lumbar interbody fusion with an autogenous iliac crest bone graft in the treatment of pyogenic spondylodiscitis. AB - There are few reports on the treatment of pyogenic lumbar spondylodiscitis through the posterior approach using a single incision. Between October 1999 and March 2003 we operated on 18 patients with pyogenic lumbar spondylodiscitis. All underwent posterior lumbar interbody fusion using an autogenous bone graft from the iliac crest and pedicle screws via a posterior approach. The clinical outcome was assessed using the Frankel neurological classification and the criteria of Kirkaldy-Willis. Under the Frankel classification, two patients improved by two grades (C to E), 11 by one grade, and five showed no change. The Kirkaldy-Willis functional outcome was excellent in five patients, good in ten and fair in three. Bony union was confirmed six months after surgery in 17 patients, but in one patient this was not achieved until two years after operation. The mean lordotic angle before operation was 20 degrees (-2 degrees to 42 degrees ) and the mean lordotic angle at the final follow-up was 32.5 degrees (17 degrees to 44 degrees ). Two patients had a superficial wound infection and two a transient root injury. Posterior lumbar interbody fusion with an autogenous iliac crest bone graft and pedicle screw fixation via a posterior approach can provide satisfactory results in pyogenic spondylodiscitis. PMID- 16720772 TI - Spinal injuries in Irish rugby: a ten-year review. AB - This study assessed the frequency of acute injury to the spinal cord in Irish Rugby over a period of ten years, between 1995 and 2004. There were 12 such injuries; 11 were cervical and one was thoracic. Ten occurred in adults and two in schoolboys. All were males playing Rugby Union and the mean age at injury was 21.6 years (16 to 36). The most common mechanism of injury was hyperflexion of the cervical spine and the players injured most frequently were playing at full back, hooker or on the wing. Most injuries were sustained during the tackle phase of play. Six players felt their injury was preventable. Eight are permanently disabled as a result of their injury. PMID- 16720773 TI - Determinants of functional outcome after simple and complex acetabular fractures involving the posterior wall. AB - We have evaluated the functional, clinical and radiological outcome of patients with simple and complex acetabular fractures involving the posterior wall, and identified factors associated with an adverse outcome. We reviewed 128 patients treated operatively for a fracture involving the posterior wall of the acetabulum between 1982 and 1999. The Musculoskeletal Functional Assessment and Short-Form 36 scores, the presence of radiological arthritis and complications were assessed as a function of injury, treatment and clinical variables. The patients had profound functional deficits compared with the normal population. Anatomical reduction alone was not sufficient to restore function. The fracture pattern, marginal impaction and residual displacement of > 2 mm were associated with the development of arthritis, which related to poor function and the need for hip replacement. It may be appropriate to consider immediate total hip replacement for patients aged > 50 years with marginal impaction and comminution of the wall, since 7 of 13 (54%) of these required early hip replacement. PMID- 16720774 TI - An alternative method for the treatment of nonunion of the tibia with bone loss. AB - In this retrospective study we evaluated the method of acute shortening and distraction osteogenesis for the treatment of tibial nonunion with bone loss in 17 patients with a mean age of 36 years (10 to 58). The mean bone loss was 5.6 cm (3 to 10). In infected cases, we performed the treatment in two stages. The mean follow-up time was 43.5 months (24 to 96). The mean time in external fixation was 8.0 months (4 to 13) and the mean external fixator index was 1.4 months/cm (1.1 to 1.8). There was no recurrence of infection. The bone evaluation results were excellent in 16 patients and good in one, while functional results were excellent in 15 and good in two. The complication rate was 1.2 per patient. We conclude that acute shortening and distraction osteogenesis is a safe, reliable and successful method for the treatment of tibial nonunion with bone loss, with a shorter period of treatment and lower rate of complication. PMID- 16720775 TI - Survivorship and clinical outcome of modular endoprosthetic reconstruction for neoplastic disease of the lower limb. AB - We reviewed retrospectively the results in 211 consecutive patients who had undergone limb salvage for bone neoplasia with endoprosthetic reconstruction of the proximal femur (96), distal femur (78), proximal tibia (30) and total femur (7). Their mean age was 50 years (11 to 86) and the mean follow-up period was 37.3 months (1 to 204). A total of 35 (16.6%) prostheses failed. Overall, implant survival was 78% (95% confidence interval (CI) 0.29 to 0.54) at five years, 60% (95% CI 0.93 to 2.35) at ten years and 60% (95% CI 1.27 to 3.88) at 15 years. Survivorship of the limb was 97.6% (95% CI 1.73 to 3.35) at ten years. The gender, age, diagnosis and location of the tumour were not prognostic variables for failure. Modular endoprosthetic replacement in the lower limb is a durable long-term reconstructive option, with the implants generally outlasting the patient. PMID- 16720776 TI - Trampoline injuries in children. AB - We reviewed the records of children referred to our hospital between April and September 2005 who had been injured whilst trampolining. Of 88 such children there were 33 boys and 55 girls with a mean age of 8 years 6 months (2 years 4 months to 15 years 9 months). Most of the injuries (53; 60%) occurred when bouncing and 34 (39%) were secondary to falls from the trampoline. The cause of injury was unknown in one child. The injured child was supervised in only 35 cases (40%). In 31 (35%) cases, the injury was related to the presence of others on the trampoline. A total of 36 (40%) children required surgery. Fractures of the upper limbs occurred in 62 cases (70%). Injuries related to the recreational use of trampolines are a significant cause of childhood injury. Our results suggest strongly that there is a need for clear guidelines on safe and responsible use of domestic trampolines. PMID- 16720777 TI - Oblique pelvic osteotomy in the exstrophy/epispadias complex. AB - We reviewed retrospectively 45 patients (46 procedures) with bladder exstrophy treated by bilateral oblique pelvic osteotomy in conjunction with genitourinary repair. The operative technique and post-operative management with or without external fixation are described. A total of 21 patients attended a special follow up clinic and 24 were interviewed by telephone. The mean follow-up time was 57 months (24 to 108). Of the 45 patients, 42 reported no pain or functional disability, although six had a waddling gait and two had marked external rotation of the hip. Complications included three cases of infection and loosening of the external fixator requiring early removal with no deleterious effect. Mid-line closure failed in one neonate managed in plaster. This patient underwent a successful revision procedure several months later using repeat osteotomies and external fixation. The percentage pubic approximation was measured on anteroposterior radiographs pre-operatively, post-operatively and at final follow up. The mean approximation was 37% (12% to 76%). It varied markedly with age and was better when external fixation was used. The wide range reflects the inability of the anterior segment to develop naturally in spite of close approximation at operation. We conclude that bilateral oblique pelvic osteotomy with or without external fixation is useful in the management of difficult primary closure in bladder exstrophy, failed primary closure and secondary reconstruction. PMID- 16720778 TI - Methicillin-resistant Staphylococcus aureus in an Irish orthopaedic centre: a five-year analysis. AB - This prospective five-year study analyses the impact of methicillin-resistant Staphylococcus aureus (MRSA) on an Irish orthopaedic unit. We identified 318 cases of MRSA, representing 0.76% of all admissions (41,971). A total of 240 (76%) cases were colonised with MRSA, while 120 (37.7%) were infected. Patients were admitted from home (218; 68.6%), nursing homes (72; 22.6%) and other hospitals (28; 8.8%). A total of 115 cases (36.6%) were colonised or infected on admission. Many patients were both colonised and infected at some stage. The length of hospital stay was almost trebled because of the presence of MRSA infection. Encouragingly, overall infection rates have not risen significantly over the five years of the study despite increased prevalence of MRSA. However, the financial burden of MRSA is increasing, highlighting the need for progress in understanding how to control this resistant pathogen more effectively. PMID- 16720779 TI - Methicillin-resistant Staphylococcus aureus on orthopaedic wards: incidence, spread, mortality, cost and control. AB - We examined the rates of infection and colonisation by methicillin-resistant Staphylococcus aureus (MRSA) between January 2003 and May 2004 in order to assess the impact of the introduction of an MRSA policy in October 2003, which required all admissions to be screened. Emergency admissions were treated prophylactically and elective beds ring-fenced. A total of 5,594 admissions were cross-referenced with 22,810 microbiology results. The morbidity, mortality and cost of managing MRSA-carrying patients, with a proximal fracture of the femur were compared, in relation to age, gender, American Society of Anaesthesiologists grade and residential status, with a group of matched controls who were MRSA-negative. In 2004, we screened 1795 of 1,796 elective admissions and MRSA was found in 23 (1.3%). We also screened 1,122 of 1,447 trauma admissions and 43 (3.8%) were carrying MRSA. All ten ward transfers were screened and four (40%) were carriers (all p < 0.001). The incidence of MRSA in trauma patients increased by 2.6% per week of inpatient stay (r = 0.97, p < 0.001). MRSA developed in 2.9% of trauma and 0.2% of elective patients during that admission (p < 0.001). The implementation of the MRSA policy reduced the incidence of MRSA infection by 56% in trauma patients (1.57% in 2003 (17 of 1,084) to 0.69% in 2004 (10 of 1,447), p = 0.035). Infection with MRSA in elective patients was reduced by 70% (0.56% in 2003 (7 of 1,257) to 0.17% in 2004 (3 of 1,806), p = 0.06). The cost of preventing one MRSA infection was 3,200 pounds. Although colonisation by MRSA did not affect the mortality rate, infection by MRSA more than doubled it. Patients with proximal fractures of the femur infected with MRSA remained in hospital for 50 extra days, had 19 more days of vancomycin treatment and 26 more days of vacuum-assisted closure therapy than the matched controls. These additional costs equated to 13,972 pounds per patient. From this experience we have been able to describe the epidemiology of MRSA, assess the impact of infection-control measures on MRSA infection rates and determine the morbidity, mortality and economic cost of MRSA carriage on trauma and elective orthopaedic wards. PMID- 16720780 TI - Resurfacing arthroplasty of the hip in osteopetrosis. AB - A 34-year-old woman with a benign form of osteopetrosis developed osteoarthritis of the hip. In order to avoid the difficulties associated with inserting the femoral component of a conventional total hip arthroplasty, a hybrid metal-on metal resurfacing was performed. There were several technical challenges associated with the procedure, including the sizing of the component, press-fit fixation of the acetabular component and femoral head preparation, as well as trying to avoid a fracture. No surgical complication occurred. After more than a year following surgery, the patient showed excellent clinical function and remained satisfied with the outcome. We conclude that the hybrid metal-on-metal resurfacing arthroplasty represents a valuable option for the treatment of patients with osteopetrosis and secondary hip osteoarthritis. PMID- 16720781 TI - The early response to major trauma and intramedullary nailing. AB - The stress response to trauma is the summation of the physiological response to the injury (the 'first hit') and by the response to any on-going physiological disturbance or subsequent trauma surgery (the 'second hit'). Our animal model was developed in order to allow the study of each of these components of the stress response to major trauma. High-energy, comminuted fracture of the long bones and severe soft-tissue injuries in this model resulted in a significant tropotropic (depressor) cardiovascular response, transcardiac embolism of medullary contents and activation of the coagulation system. Subsequent stabilisation of the fractures using intramedullary nails did not significantly exacerbate any of these responses. PMID- 16720782 TI - Factors affecting the cohesion of impaction bone graft. AB - The role of bone-graft extenders in impaction revision surgery is becoming increasingly important. Tricalcium phosphate and hydroxyapatite have been shown to be both biocompatible and osteoconductive, yet many surgeons remain reluctant to use them. The difficulty in handling bone-graft extenders can be partly alleviated by using porous particles and adding clotted blood. In an in vitro model we measured the cohesive properties of various impaction graft mixes. Several factors were evaluated including the use of pure bone graft compared with mixes with extender, washing the bone and the addition of clotted blood. Our findings showed that pure allograft bone particles had significantly higher cohesion than when mixed with extender (p < 0.001). Washing had no effect on cohesion. The addition of clotted blood significantly increased the cohesion of both pure bone (p < 0.019) and mixes with pure bone and with porous graft extender (p < 0.044). PMID- 16720783 TI - Reconstruction of femoral defects in revision hip surgery: risk of fracture and stem migration after impaction bone grafting. AB - The use of impaction bone grafting during revision arthroplasty of the hip in the presence of cortical defects has a high risk of post-operative fracture. Our laboratory study addressed the effect of extramedullary augmentation and length of femoral stem on the initial stability of the prosthesis and the risk of fracture. Cortical defects in plastic femora were repaired using either surgical mesh without extramedullary augmentation, mesh with a strut graft or mesh with a plate. After bone impaction, standard or long-stem Exeter prostheses were inserted, which were tested by cyclical loading while measuring defect strain and migration of the stem. Compared with standard stems without extramedullary augmentation, defect strains were 31% lower with longer stems, 43% lower with a plate and 50% lower with a strut graft. Combining extramedullary augmentation with a long stem showed little additional benefit (p = 0.67). The type of repair did not affect the initial stability. Our results support the use of impaction bone grafting and extramedullary augmentation of diaphyseal defects after mesh containment. PMID- 16720784 TI - Removal of acetabular bone in resurfacing arthroplasty of the hip. PMID- 16720785 TI - Removal of acetabular bone in resurfacing arthroplasty of the hip. PMID- 16720786 TI - Removal of acetabular bone in resurfacing arthroplasty of the hip. PMID- 16720787 TI - The premature failure of the Charnley Elite-Plus stem. PMID- 16720788 TI - Removal of acetabular bone in resurfacing arthroplasty of the hip. PMID- 16720789 TI - Blast-related traumatic brain injury: what is known? PMID- 16720790 TI - Behavioral neurology and neuropsychiatry is a subspecialty. PMID- 16720791 TI - Regional alpha-synuclein aggregation, dopaminergic dysregulation, and the development of drug-related visual hallucinations in Parkinson's disease. AB - Visual hallucinations in Parkinson's disease are usually treatment-related and occur in at least 30% of patients. Although their clinical and epidemiological features have been extensively reviewed, their etiopathogenesis remains a matter of debate. Based on the current evidence available, this review suggests that regional neurodegeneration of the ventral dopaminergic pathway, as evident in the aggregation of the protein alpha-synuclein, is the main event linked to the development of visual hallucinations in Parkinson's disease. Denervation supersensitivity of dopaminergic receptors in ventral striatal and mesocorticolimbic areas as well as defective synaptic buffering ability due to the loss of dopaminergic presynaptic terminals and dopamine transporter may be among the key factors leading to visual hallucinations in Parkinson's disease. PMID- 16720792 TI - The neuropsychiatry of Niemann-Pick type C disease in adulthood. AB - Psychotic symptoms occur in a variety of medical and neurological conditions. The authors describe three young men with a variant form of Niemann-Pick type C disease, a neurodegenerative disorder related to abnormal intracellular cholesterol metabolism, who presented with psychosis in early adulthood. Two patients were treated for schizophrenia for many years prior to a diagnosis of Niemann-Pick type C. The cases presented in this article illustrate the role of changes in both white and gray matter structures in psychosis, and, like the assessments of other neurodevelopmental disorders that predispose toward psychotic presentations, shed light on the underlying pathophysiology of major mental disorders. PMID- 16720793 TI - Dementia with mood symptoms in a fragile X premutation carrier with the fragile X associated tremor/ataxia syndrome: clinical intervention with donepezil and venlafaxine. AB - The authors present a case of a patient with dementia with mood symptoms and multiple neurological manifestations of fragile X-associated tremor/ataxia syndrome (FXTAS). Despite a gradually deteriorating neurological course, he was managed for 2 years with combination therapy of donepezil and venlafaxine, which resulted in improvement and relative stabilization of his psychiatric status. Psychiatrists are hereby alerted to the description of a novel dementia syndrome that may respond to pharmacological intervention commonly used for other dementias. PMID- 16720795 TI - Cognitive reserve and the relationship between depressive symptoms and awareness of deficits in dementia. AB - Depression and low cognitive reserve are linked to impaired awareness in dementia, although their relative contributions are unknown. The authors investigated the impact of depressive symptoms and cognitive reserve on awareness in questionable and mild dementia. Sixty-six patients completed measures of cognitive reserve, awareness, and depressive symptoms. Cognitive reserve and its interaction with depressive symptoms accounted for significant proportions of variance in awareness; depressive symptoms alone did not. Depressive symptoms and awareness were positively related in high, but not low, cognitive reserve groups. Findings suggest cognitive reserve moderates the depression/awareness relationship in dementia, which may explain previously discrepant findings. PMID- 16720794 TI - Donepezil effects on cerebral blood flow in older adults with mild cognitive deficits. AB - Cerebral blood flow (CBF) was assessed during a verbal recall task using [(15)O]water positron emission tomography (PET) in older adults with mild cognitive deficits participating in a placebo-controlled donepezil trial. The placebo group demonstrated reduced CBF in the left frontal and temporal regions over the 6-month period, while those receiving donepezil did not. The placebo group's performance did not change on a list-learning task, while the donepezil group's performance improved, despite having had lower performance at intake. These findings suggest that donepezil treatment may be associated with a relative maintenance of CBF and improved list-learning. PMID- 16720796 TI - White matter changes associated with psychotic symptoms in Alzheimer's disease patients. AB - This study explored the relationship between white matter changes seen on magnetic resonance imaging (MRI) and neuropsychiatric symptoms of Alzheimer's disease patients. Fifty-five probable Alzheimer's disease patients were assessed with Behavioral Rating Scale for Dementia (BRSD) and MRI. White matter changes in the bilateral frontal or parieto-occipital region and left basal ganglia significantly corresponded with the score of the Psychotic Symptoms subscale of BRSD. Secondary analyses revealed that white matter changes were not associated with paranoid delusion and hallucination, but only with delusional misidentification. Our results suggest that white matter changes in Alzheimer's disease patients probably contribute to the development of specific psychotic symptoms, namely delusional misidentification. PMID- 16720797 TI - Compromised memory function in schizophrenia and temporal lobe epilepsy. AB - The purpose of this study was to investigate compromised memory function of schizophrenia patients in comparison with temporal lobe epilepsy patients, whose memory impairments result from a clear lesion. The authors hypothesized that schizophrenia patients would show poorer immediate and delayed recall performances in verbal and visual memory tasks. The study sample consisted of a healthy comparison group of 30 subjects and three patient groups comprising 76 schizophrenia patients, 93 left temporal lobe epilepsy patients, and 72 right temporal lobe epilepsy patients. The authors assessed immediate recall, delayed recall, and delayed retention. Tasks were subdivided into two categories (easy and difficult), and then patient memory dysfunction was compared among the memory tests. The authors observed material-specific memory impairment, where the left temporal lobe epilepsy group showed severe verbal memory impairment and the right temporal lobe epilepsy group showed severe visual memory impairment. A moderate impairment was found in immediate and delayed verbal memory in schizophrenia patients, and the impairment of visual memory was amplified with delayed recall. Such a result can be interpreted not only as a generalized cognitive deficit, but also as an integrative dysfunction involving the mesial temporal and frontal lobes in the left and right hemispheres, whereby the lesion site cannot be determined selectively. Our results show that the selection of a memory task that cannot be influenced by verbal mediation is very important for analyzing memory dysfunction in schizophrenia patients. PMID- 16720799 TI - Neurocognition in depression: patients on and off medication versus healthy comparison subjects. AB - Patients with depression have neuropsychological deficits in attention, memory, psychomotor speed, processing speed, and executive function. It is not clear, however, whether neurocognition in depression is impaired in a global or nonspecific way or if specific cognitive domains are selectively impaired. This naturalistic cross-sectional study employed a computerized neurocognitive screening battery to evaluate 38 depressed, drug-free patients, compared to 31 patients who responded to antidepressant monotherapy and to 69 healthy comparison subjects. There was evidence for global neuropsychological impairment in untreated depressed patients. In patients who had been successfully treated, performance was improved but not normalized. There was also evidence for specific depression-related deficits in executive function and processing speed but not in memory, psychomotor speed, or reaction time. Although depressed patients have global neurocognitive impairments, deficits in certain cognitive domains are more important than in others. In particular, impairments are noted in tests of executive control and in tests that demand effortful attention. Information processing speed is also impaired but not reaction time. Computerized testing in the clinic setting demonstrates a range of neurocognitive problems in patients with depression. These problems may have a bearing on treatment and outcome. PMID- 16720798 TI - Variation in neurophysiological function and evidence of quantitative electroencephalogram discordance: predicting cocaine-dependent treatment attrition. AB - Cocaine treatment trials suffer from a high rate of attrition. We examined pretreatment neurophysiological factors to identify participants at greatest risk. Twenty-five participants were divided into concordant and discordant groups following electroencephalogram (EEG) measures recorded prior to a double-blind, placebo-controlled treatment trial. Three possible outcomes were examined: successful completion, dropout, and removal. Concordant (high perfusion correlate) participants had an 85% rate of successful completion, while discordant participants had a 15% rate of successful completion. Twenty-five percent of dropouts and 50% of participants removed were discordant (low perfusion correlate), while only 25% of those who completed were discordant. Failure to complete the trial was not explained by depression, craving, benzoylecgonine levels or quantitative electroencephalogram (QEEG) power; thus cordance may help identify attrition risk. PMID- 16720800 TI - Cognitive function and dissociative disorder status among veteran subjects with chronic posttraumatic stress disorder: a preliminary study. AB - Twenty-seven veteran subjects with chronic posttraumatic stress disorder (PTSD) were evaluated for dissociative disorders. Ten subjects met criteria for one or more dissociative disorders, and 17 subjects did not meet dissociative disorder criteria. Neurocognitive profiles of the two groups differed in several areas, with veterans meeting diagnostic criteria for both PTSD and a dissociative disorder and demonstrating considerably greater deficits in attention, autobiographical memory, and verbal memory than PTSD subjects without comorbid dissociative disorder diagnosis. PMID- 16720801 TI - Apolipoprotein E genotype and odor identification in schizophrenia. AB - The authors examined Apolipoprotein E (ApoE) genotype frequencies and unirhinal odor identification in 28 schizophrenia patients and 26 healthy comparison subjects. No significant associations between ApoE status and olfaction were observed in either diagnostic group. The authors concluded that olfactory deficits in schizophrenia do not appear to be mediated by the ApoE allele. PMID- 16720802 TI - Topiramate effect in catatonia: a case series. AB - The authors describe four cases of catatonia in which topiramate treatment was used. Commonly effective therapies, including benzodiazepines and divalproex, were proven refractory. In all four cases, subjects experienced complete remission of catatonic symptoms and tolerated treatment well. In one case, all psychotropic medications were discontinued because the patient became delirious. The delirium resolved after discovery and treatment of a urinary tract infection. Catatonic agitation relapsed when topiramate was withdrawn but remitted again when topiramate and lorazepam therapy was restored. In two cases, continued topiramate therapy was accompanied by sustained remission. These case reports present a novel approach to the treatment of catatonia. PMID- 16720803 TI - Mood and behavioral dysfunction with opsoclonus-myoclonus ataxia. AB - Opsoclonus-myoclonus ataxia syndrome is a paraneoplastic syndrome of cerebellar damage associated with neuroblastoma. The authors assessed psychiatric symptoms of opsoclonus-myoclonus ataxia syndrome in 17 children, who were 16 months to 12(1/2) years of age. Psychiatric symptoms examined included disruptive behavior, affective dysregulation, irritability, impulsivity, cognitive impairment, and poor attention. PMID- 16720804 TI - Buspirone for stereotypic movements in elderly with cognitive impairment. AB - Repetitive and stereotypic behavioral disturbances in patients with dementia are common; however, little is known regarding successful treatments. The authors describe six cases of elderly cognitively impaired patients exhibiting repetitive and stereotypic behaviors who were treated successfully with buspirone. The cases demonstrate that buspirone may be an effective and safe treatment for patients with dementia who demonstrate repetitive and stereotypic behavior disorders. PMID- 16720805 TI - Clozapine-induced tardive dyskinesia and hypothyroidism. PMID- 16720806 TI - Clozapine and ziprasidone: a useful combination in patients with treatment resistant schizophrenia. PMID- 16720807 TI - Insulinoma in differential diagnosis of seizure disorder. PMID- 16720808 TI - Antipsychotic treatment improves outcome in herpes simplex encephalitis: a case report. PMID- 16720809 TI - Modafinil-induced reversible hyperkinetic nondystonic movement disorder in a patient with major depressive disorder. PMID- 16720811 TI - The cover. New Mexico Skies. PMID- 16720812 TI - A piece of my mind. Copy-and-paste. PMID- 16720813 TI - Rx for obesity: eat less, exercise more, and--maybe--get more sleep. PMID- 16720814 TI - Chronic homeless in intervention spotlight. PMID- 16720815 TI - Genome provides clues on addiction. PMID- 16720816 TI - Marburg vaccine shows promise: offers postexposure protection in monkeys. PMID- 16720817 TI - Low-molecular-weight heparin vs unfractionated heparin in acute coronary syndromes. PMID- 16720818 TI - Trojan horse hypothesis: inhaled airborne particles, lipid bullets, and atherogenesis. PMID- 16720819 TI - Research in bariatric surgery. PMID- 16720820 TI - Research in bariatric surgery. PMID- 16720821 TI - Screening colonoscopy in very elderly patients: prevalence of neoplasia and estimated impact on life expectancy. AB - CONTEXT: Current guidelines do not include an upper age cutoff for colorectal cancer screening with colonoscopy. Although the prevalence of colonic neoplasia increases with age, life expectancy decreases. Thus, the benefit of screening colonoscopy in very elderly patients may be limited. OBJECTIVE: To compare estimated life-years saved with screening colonoscopy in very elderly vs younger persons. DESIGN, SETTING, AND PARTICIPANTS: Cross-sectional study conducted among 1244 asymptomatic individuals in 3 age groups (50-54 years [n = 1034], 75-79 years [n = 147], and > or =80 years [n = 63]) who underwent screening colonoscopy at a US teaching hospital and clinic. MAIN OUTCOME MEASURES: Prevalence of various types of colon neoplasia; estimated gain in life expectancy, calculated as life expectancy--(life expectancy during polyp lag time + life expectancy after colorectal cancer diagnosis); and comparison of mean gain in life expectancy across the 3 groups. Life expectancy and mortality data were derived from life tables, previous studies, and national databases. RESULTS: The prevalence of neoplasia was 13.8% in the 50- to 54-year-old group, 26.5% in the 75- to 79-year-old group, and 28.6% in the group aged 80 years or older. Despite higher prevalence of neoplasia in elderly patients, mean extension in life expectancy was much lower in the group aged 80 years or older than in the 50- to 54-year-old group (0.13 vs 0.85 years). In sensitivity analysis, with longer polyp lag times the mean extension in life expectancy decreased more in the elderly than in the younger patients; alternatively, if it was assumed that a smaller proportion of adenomas progress to colorectal cancer, the mean extension in life expectancy decreased less in the elderly than in the younger patients. CONCLUSIONS: Even though prevalence of neoplasia increases with age, screening colonoscopy in very elderly persons (aged > or =80 years) results in only 15% of the expected gain in life expectancy in younger patients. Screening colonoscopy in very elderly patients should be performed only after careful consideration of potential benefits, risks, and patient preferences. PMID- 16720822 TI - Risk of developing colorectal cancer following a negative colonoscopy examination: evidence for a 10-year interval between colonoscopies. AB - CONTEXT: Limited evidence exists to guide the optimal frequency of repeat endoscopic examination for colorectal cancer screening after a negative colonoscopy. OBJECTIVE: To determine the duration and magnitude of the risk of developing colorectal cancer following performance of a negative colonoscopy. DESIGN, SETTING, AND PATIENTS: Population-based retrospective analysis of individuals whose colonoscopy evaluations did not result in a diagnosis of colorectal neoplasia. Patients who had been evaluated between April 1, 1989, and December 31, 2003, were identified using Manitoba Health's physician billing claims database (N = 35 975). Standardized incidence ratios (SIRs) were calculated to compare colorectal cancer incidence in our cohort with colorectal cancer incidence in the provincial population. Stratified analysis was performed to determine the duration of the reduced risk. Patients with a history of colorectal cancer prior to the index colonoscopy, inflammatory bowel disease, resective colorectal surgery, and lower gastrointestinal endoscopy within the 5 years before the index colonoscopy were excluded. Cohort members were followed up from the time of the index colonoscopy until diagnosis of colorectal cancer, death, out-migration from Manitoba, or end of the study period on December 31, 2003. MAIN OUTCOME MEASURE: Incidence of colorectal cancer. RESULTS: A negative colonoscopy was associated with SIRs of 0.69 (95% confidence interval [CI], 0.59 0.81) at 6 months, 0.66 (95% CI, 0.56-0.78) at 1 year, 0.59 (95% CI, 0.48-0.72) at 2 years, 0.55 (95% CI, 0.41-0.73) at 5 years, and 0.28 (95% CI, 0.09-0.65) at 10 years. The proportion of colorectal cancer located in the right side of the colon was significantly higher in the colonoscopy cohort than the rate in the Manitoba population (47% vs 28%; P<.001). CONCLUSIONS: The risk of developing colorectal cancer remains decreased for more than 10 years following the performance of a negative colonoscopy. There is a need to improve the early detection rate of right-sided colorectal neoplasia in usual clinical practice. PMID- 16720823 TI - Cost-effectiveness of screening BRCA1/2 mutation carriers with breast magnetic resonance imaging. AB - CONTEXT: Women with inherited BRCA1/2 mutations are at high risk for breast cancer, which mammography often misses. Screening with contrast-enhanced breast magnetic resonance imaging (MRI) detects cancer earlier but increases costs and results in more false-positive scans. OBJECTIVE: To evaluate the cost effectiveness of screening BRCA1/2 mutation carriers with mammography plus breast MRI compared with mammography alone. DESIGN, SETTING, AND PATIENTS: A computer model that simulates the life histories of individual BRCA1/2 mutation carriers, incorporating the effects of mammographic and MRI screening was used. The accuracy of mammography and breast MRI was estimated from published data in high risk women. Breast cancer survival in the absence of screening was based on the Surveillance, Epidemiology and End Results database of breast cancer patients diagnosed in the prescreening period (1975-1981), adjusted for the current use of adjuvant therapy. Utilization rates and costs of diagnostic and treatment interventions were based on a combination of published literature and Medicare payments for 2005. MAIN OUTCOME MEASURES: The survival benefit, incremental costs, and cost-effectiveness of MRI screening strategies, which varied by ages of starting and stopping MRI screening, were computed separately for BRCA1 and BRCA2 mutation carriers. RESULTS: Screening strategies that incorporate annual MRI as well as annual mammography have a cost per quality-adjusted life-year (QALY) gained ranging from less than 45,000 dollars to more than 700,000 dollars, depending on the ages selected for MRI screening and the specific BRCA mutation. Relative to screening with mammography alone, the cost per QALY gained by adding MRI from ages 35 to 54 years is 55,420 dollars for BRCA1 mutation carriers, 130,695 dollars for BRCA2 mutation carriers, and 98,454 dollars for BRCA2 mutation carriers who have mammographically dense breasts. CONCLUSIONS: Breast MRI screening is more cost-effective for BRCA1 than BRCA2 mutation carriers. The cost-effectiveness of adding MRI to mammography varies greatly by age. PMID- 16720825 TI - A 76-year-old man with macular degeneration. PMID- 16720824 TI - Trends in the association of poverty with overweight among US adolescents, 1971 2004. AB - CONTEXT: Prevalence of adolescent overweight in the United States has increased substantially during the past 3 decades. Whether socioeconomic disparities in adolescent overweight increased, decreased, or remained constant during this period is not known. OBJECTIVE: To examine trends in adolescent overweight from 1971 to 2004 by family poverty status, as well as trends in potentially relevant eating and physical activity behaviors. DESIGN, SETTING, AND PARTICIPANTS: Four cross-sectional, nationally representative surveys (US National Health and Nutrition Examination Surveys [NHANES] of 1971-1974, 1976-1980, 1988-1994, and 1999-2004) were examined for trends in the prevalence of overweight among adolescents aged 12 to 17 years by family poverty status. MAIN OUTCOME MEASURES: Prevalence of adolescent overweight, defined as body mass index at or above the 95th percentile for age and sex in the 2000 Centers for Disease Control and Prevention growth charts. Intermediate outcomes were physical inactivity in the past 30 days, proportion of caloric intake from sweetened beverages (24-hour recall), and whether respondent skipped breakfast (24-hour recall). RESULTS: Trends in the association of adolescent overweight with family poverty differed by age stratum (P = .01). In 12- to 14-year-old adolescents, prevalence did not significantly differ by family poverty status in any of the surveys; however, among non-Hispanic black adolescents, overweight prevalence increased faster in nonpoor vs poor families. In contrast, a widening disparity that disfavored adolescents from poor families was present in the 15- to 17-year-old adolescents. This trend was similar among male, female, non-Hispanic white, and non-Hispanic black adolescents, resulting in an overall prevalence of overweight in 1999-2004 more than 50% higher among adolescents in poor vs nonpoor families (23.3% vs 14.4%, respectively; P<.001). Additional analyses suggest that physical inactivity, sweetened beverage consumption, and skipping breakfast may contribute to these disparities. CONCLUSIONS: Trends of increasing overweight showed a greater impact in families living below the poverty line vs not living below the poverty line among older (15-17 years) but not younger (12-14 years) adolescents. Furthermore, physical inactivity, high consumption of sweetened beverages, and breakfast skipping may be candidate targets for prevention programs aimed at reducing this recently emerged disparity. PMID- 16720826 TI - Protecting special interests in the name of "good science". PMID- 16720827 TI - Screening for colorectal cancer by colonoscopy: adding to the evidence. PMID- 16720828 TI - JAMA patient page. Age-related macular degeneration. PMID- 16720829 TI - The heat sensitivity of cytokine-inducing effect of lipopolysaccharide. AB - Heat inactivation by boiling has been widely used as a criterion to determine whether the observed effects of a protein preparation are a result of lipopolysaccharide (LPS) contamination. However, the heat sensitivity of LPS cytokine-inducing activity has not been characterized. In the current study, we demonstrated that the endotoxin activity, i.e., Limulus amebocyte lysate-gelating activity, and the tumor necrosis factor alpha (TNF-alpha)-inducing activity of LPS (Escherichia coli K-12 JM83, K-12 LCD25, and F583) were sensitive to boiling. Heat treatment by boiling for 15 min was sufficient to inactivate approximately 90% of the LPS TNF-alpha-inducing activity. The heat-induced inactivation of LPS activities was not a result of adherence of boiled LPS to the wall of the container, i.e., polypropylene tubes, or aggregation of boiled LPS. In addition, boiled LPS retained its ability to bind polymyxin B. The presence of protein (ovalbumin) in LPS did not affect the heat sensitivity of LPS. Conversely, boiling reduced the size of LPS aggregates as determined by electrophoresis using native polyacrylamide gel. Likewise, the TNF-alpha-inducing activity of diphosphoryl lipid A (DPLA) was also sensitive to boiling. Thin-layer chromatographic analysis of boiled DPLA revealed that the heat-induced inactivation of DPLA TNF-alpha-inducing activity was not a result of its conversion to monophosphoryl lipid A. We conclude that the TNF-alpha-inducing activity of LPS and DPLA is sensitive to boiling and suggest that heat sensitivity as an indicator of whether the observed effects of a protein preparation are a result of LPS contamination should be used with caution. PMID- 16720830 TI - Evaluation of the safety and pharmacodynamics of Hematide, a novel erythropoietic agent, in a phase 1, double-blind, placebo-controlled, dose-escalation study in healthy volunteers. AB - Hematide is an investigational pegylated synthetic peptide that stimulates erythropoiesis in animal models and is being developed for the treatment of anemia associated with chronic renal failure and cancer. This study evaluated the safety and pharmacodynamics of single, intravenous doses (0.025, 0.05, and 0.1 mg/kg) of Hematide in 28 healthy male volunteers. All doses of Hematide were well tolerated, with safety profiles similar to those of placebo. Hematide showed a dose-dependent increase in reticulocytes. The 0.1-mg/kg dose was associated with a statistically significant increase in hemoglobin (Hgb) from baseline compared to the placebo group (13.6 +/- 3.9 g/L [1.36 +/- 0.39 g/dL] versus 3.9 +/- 3.8 g/L [0.39 +/- 0.38 g/dL]; P < .001) that was sustained for longer than 1 month. These results support phase 2 studies in patients with anemia associated with chronic kidney disease or cancer and suggest that Hematide administered as infrequently as once a month may result in a sustained elevation of Hgb levels. (Please note that Hematide is a proposed trade name; the compound does not yet have a nonproprietary name.). PMID- 16720831 TI - Thrombin-induced endothelial microparticle generation: identification of a novel pathway involving ROCK-II activation by caspase-2. AB - Thrombin exerts pleiotropic effects on endothelial cells, including the release of microparticles (EMPs) that disseminate and exchange information with vascular cells. Nevertheless, the mechanisms leading to their generation are not elucidated. We performed microarray analysis to identify genes involved in EMP release by the endothelial cell line HMEC-1 in response to thrombin. We identified a group of genes linked to the cytoskeleton reorganization family. Among these, the Rho-kinase ROCK-II presented a high transcription rate. ROCK-I, another Rho-kinase isoform, was not modulated by thrombin. Pharmacologic inhibition of Rho-kinases or specific depletion of ROCK-II by short interfering (si) RNA inhibited thrombin-induced EMP release. In contrast, ROCK-I mRNA silencing did not modify EMP generation by thrombin. Exposure of HMEC-1 to thrombin in presence of the caspase-2 selective inhibitor Z-VDVAD-FMK prevented ROCK-II cleavage and inhibited the thrombin-induced EMP release. These events were observed in absence of cell death. Our data clearly identified ROCK-II as a target of thrombin in EMP generation. They indicated that the 2 Rho-kinases did not share identical functions. The involvement of caspase-2 in ROCK-II activation independently of cell death points out a novel signaling pathway that emphasizes the proteolytic activity of caspase in EMP generation in response to cell activation. PMID- 16720832 TI - Impaired megakaryocytopoiesis in type 2B von Willebrand disease with severe thrombocytopenia. AB - In type 2B von Willebrand disease, there is spontaneous binding of mutated von Willebrand factor (VWF) multimers to platelets. Here we report a family in which severe thrombocytopenia may also be linked to abnormal megakaryocytopoiesis. A heterozygous mutation in the VWF A1 domain gave a R1308P substitution in an interactive site for glycoprotein Ibalpha (GPIbalpha). Electron microscopy showed clusters of platelets in close contact. Binding of antibodies to the GPIbalpha N terminal domain was decreased, whereas GPIX and GPV were normally detected. In Western blotting (WB), GPIbalpha, alphaIIb, and beta3 were normally present. Proteins involved in Ca(2+) homeostasis were analyzed by quantitating platelet mRNA or by WB. Plasma membrane Ca(2+) ATPase (PMCA)-4b and type III inositol trisphosphate receptor (InsP(3)-R3) were selectively increased. The presence of degradation products of polyadenosine diphosphate (ADP)-ribose polymerase protein (PARP) suggested ongoing caspase-3 activity. These were findings typical of immature normal megakaryocytes cultured from peripheral blood CD34(+) cells with TPO. Significantly, megakaryocytes from the patients in culture produced self associated and interwoven proplatelets. Immunolocalization showed VWF not only associated with platelets, but already on the megakaryocyte surface and within internal channels. In this family, type 2B VWD is clearly associated with abnormal platelet production. PMID- 16720834 TI - Immunohistochemistry predicts nucleophosmin (NPM) mutations in acute myeloid leukemia. AB - Nucleophosmin (NPM) exon-12 mutations occur in 50% to 60% of adult acute myeloid leukemia (AML) with normal karyotype and are predictors of favorable prognosis. We evaluated bone marrow or peripheral blood samples from 450 adult patients with AML of the GIMEMA (Gruppo Italiano Malattie Ematologiche Maligne dell'Adulto)/AML12 EORTC (European Organization for Research and Treatment of Cancer) trial to (1) search for new exon-12 NPM mutations; (2) determine whether NPM immunostaining on paraffin-embedded biopsies predicts NPM mutations; and (3) investigate altered nucleocytoplasmic NPM traffic in primary AML cells. Fourteen NPM mutations, including 8 new variants, were identified. All 200 AML cases expressing cytoplasmic NPM (NPMc(+) AML) carried NPM mutations. None of the 250 cases with nucleus-restricted NPM (NPMc(-) AML) was mutated. At the C-terminus, NPM leukemic mutants carried mutations of only tryptophan 290 or of both tryptophans 288 and 290 and a new nuclear export signal (NES) motif, which appear to underlie their nuclear export. The specific Crm1/exportin-1 inhibitor leptomycin-B relocated NPM mutants from cytoplasm to nucleus of primary NPMc(+) AML cells, demonstrating that nuclear export is NES dependent. NPM mutants bound and recruited wild-type NPM into leukemic cell cytoplasm. Because alterations at C-terminus of leukemic NPM mutants are similar, immunohistochemistry detects all exon-12 NPM mutations and is a valuable, inexpensive tool in the diagnostic prognostic work-up of patients with AML with normal karyotype. PMID- 16720833 TI - Impact of the intensity of the pretransplantation conditioning regimen in patients with prior invasive aspergillosis undergoing allogeneic hematopoietic stem cell transplantation: A retrospective survey of the Infectious Diseases Working Party of the European Group for Blood and Marrow Transplantation. AB - In this retrospective study, we analyzed the outcomes of 129 patients who underwent an allogeneic hematopoietic stem cell transplantation (allo-HSCT) and had a history of probable or proven invasive aspergillosis (IA), of whom 57 (44%) received a reduced-intensity conditioning (RIC). Overall, 27 patients with IA progressed after the allo-HSCT (cumulative incidence [CumInc] at 2 years, 22%). The variables that increased the 2-year CumInc of IA progression were (1) longer duration of neutropenia after transplantation; (2) advanced status of the underlying disease; and (3) less than 6 weeks from start of systemic anti Aspergillus therapy and the allo-HSCT. In addition, (4) conventional myeloablative conditioning increased the risk of progression early after transplantation (before day 30) only, while 3 variables increased the risk beyond day 30 were (5) cytomegalovirus disease; (6) bone marrow or cord blood as source of stem cells; and (7) grades II to IV acute graft-versus-host disease (GVHD). A risk model for progression was generated, defined as low (0-1 risk factors, 6% incidence), intermediate (2-3 risk factors, 27% incidence), or high risk (> or = 3 risk factors, 72% incidence [P < .001]). These findings may help in the interpretation and design of future studies on secondary prophylaxis of IA after an allo-HSCT. PMID- 16720835 TI - Impaired "outside-in" integrin alphaIIbbeta3 signaling and thrombus stability in TSSC6-deficient mice. AB - We investigated the role of the hematopoietic-specific tetraspanin superfamily member, TSSC6, in platelet function using wild-type mice and TSSC6-deficient mice. TSSC6 is expressed on the surface of murine platelets and is up-regulated by thrombin stimulation, indicating an intracellular pool of TSSC6. Immunoprecipitation/Western blot studies reveal a constitutive physical association of TSSC6 with the integrin alpha(IIb)beta(3) complex under strong detergent conditions. In vivo evaluation of hemostasis by tail bleeding revealed increased bleeding time, volume of blood lost, and evidence of tail rebleeds in TSSC6 null mice, indicating unstable hemostasis. Using ex vivo techniques, we showed that TSSC6-deficient platelets exhibited impaired kinetics of clot retraction, platelet aggregation at lower doses of PAR-4, and collagen and platelet spreading on fibrinogen in the presence of normal integrin alpha(IIb)beta(3) expression. TSSC6-deficient platelets showed normal alpha granule secretion, normal "inside-out" integrin alpha(IIb)beta(3) signaling (fluorescein isothiocyanate [FITC]-fibrinogen and JON/A binding), and normal platelet adhesion on fibrinogen. Furthermore, we show that absence of platelet TSSC6 affects the secondary stability of arterial thrombi in vivo upon vascular injury. These data demonstrate that TSSC6 appears to regulate integrin alpha(IIb)beta(3) "outside-in" signaling events in platelets and is necessary for stability of arterial thrombi in vivo. PMID- 16720836 TI - Variations of the perforin gene in patients with autoimmunity/lymphoproliferation and defective Fas function. AB - Mutations decreasing function of the Fas death receptor cause the autoimmune lymphoproliferative syndrome (ALPS) with autoimmune manifestations, spleen/lymph node enlargement, and expansion of CD4/CD8-negative T cells. Dianzani Autoimmune Lymphoproliferative Disease (DALD) is a variant lacking this expansion. Perforin is involved in cell-mediated cytotoxicity and its biallelic mutations cause familial hemophagocytic lymphohistiocytosis (HLH). We previously described an ALPS patient carrying heterozygous mutations of the Fas and perforin genes and suggested that they concurred in ALPS. This work extends the analysis to 14 ALPS, 28 DALD, and 816 controls, and detects an N252S amino acid substitution in 2 ALPS, and an A91V amino acid substitution in 6 DALD. N252S conferred an OR = 62.7 (P = .0016) for ALPS and A91V conferred an OR = 3 (P = .016) for DALD. Copresence of A91V and variations of the osteopontin gene previously associated with DALD conferred an OR = 17 (P = .0007) for DALD. In one N252S patient, NK activity was strikingly defective in early childhood, but became normal in late childhood. A91V patients displayed lower NK activity than controls. These data suggest that perforin variations are a susceptibility factor for ALPS/DALD development in subjects with defective Fas function and may influence disease expression. PMID- 16720837 TI - Somatic activation of a conditional KrasG12D allele causes ineffective erythropoiesis in vivo. AB - Somatic activation of a conditional targeted Kras(G12D) allele induces a fatal myeloproliferative disease in mice that closely models juvenile and chronic myelomonocytic leukemia. These mice consistently develop severe and progressive anemia despite adequate numbers of clonogenic erythroid progenitors in the bone marrow and expanded splenic hematopoiesis. Ineffective erythropoiesis is characterized by impaired differentiation. These results demonstrate that endogenous levels of oncogenic Ras have cell lineage-specific effects and support efforts to modulate Ras signaling for therapy of anemia in patients with myelodysplastic syndromes and myeloproliferative disorders. PMID- 16720840 TI - Second messenger pas de deux: the coordinated dance between calcium and cAMP. AB - Dynamic calcium signaling is a well-established precept in biology. Different cell types exhibit spontaneous as well as stimulus-triggered transient changes in the concentration of intracellular calcium. Does this behavior extend to other second messengers? Optical dissection of various signal transduction pathways with fluorescent reporter molecules that enable visualization of changes in concentration of other second messengers is well under way. Recent research using technologically refined probes provides improved temporal and spatial resolution of adenosine 3',5'-monophosphate (cAMP) dynamics to generate insights into the bidirectional interplay between intracellular fluctuations of cAMP and calcium. cAMP oscillations are generated in response to hormones, and cells can recognize and differentially respond to transient versus sustained changes in this second messenger. Second messenger reporters are now available to track multiple players and so provide a dynamic picture of signaling networks. PMID- 16720839 TI - Evidence for subcomplexes in the Fanconi anemia pathway. AB - Fanconi anemia (FA) is a genomic instability disorder, clinically characterized by congenital abnormalities, progressive bone marrow failure, and predisposition to malignancy. Cells derived from patients with FA display a marked sensitivity to DNA cross-linking agents, such as mitomycin C (MMC). This observation has led to the hypothesis that the proteins defective in FA are involved in the sensing or repair of interstrand cross-link lesions of the DNA. A nuclear complex consisting of a majority of the FA proteins plays a crucial role in this process and is required for the monoubiquitination of a downstream target, FANCD2. Two new FA genes, FANCB and FANCL, have recently been identified, and their discovery has allowed a more detailed study into the molecular architecture of the FA pathway. We demonstrate a direct interaction between FANCB and FANCL and that a complex of these proteins binds FANCA. The interaction between FANCA and FANCL is dependent on FANCB, FANCG, and FANCM, but independent of FANCC, FANCE, and FANCF. These findings provide a framework for the protein interactions that occur "upstream" in the FA pathway and suggest that besides the FA core complex different subcomplexes exist that may have specific functions other than the monoubiquitination of FANCD2. PMID- 16720838 TI - Compound heterozygosity of novel missense mutations in the gamma-glutamyl carboxylase gene causes hereditary combined vitamin K-dependent coagulation factor deficiency. AB - Hereditary combined vitamin K-dependent (VKD) coagulation factor deficiency is an autosomal recessive bleeding disorder associated with defects in either the gamma carboxylase, which carboxylates VKD proteins to render them active, or the vitamin K epoxide reductase (VKORC1), which supplies the reduced vitamin K cofactor required for carboxylation. Such deficiencies are rare, and we report the fourth case resulting from mutations in the carboxylase gene, identified in a Tunisian girl who exhibited impaired function in hemostatic VKD factors that was not restored by vitamin K administration. Sequence analysis of the proposita did not identify any mutations in the VKORC1 gene but, remarkably, revealed 3 heterozygous mutations in the carboxylase gene that caused the substitutions Asp31Asn, Trp157Arg, and Thr591Lys. None of these mutations have previously been reported. Family analysis showed that Asp31Asn and Thr591Lys were coallelic and maternally transmitted while Trp157Arg was transmitted by the father, and a genomic screen of 100 healthy individuals ruled out frequent polymorphisms. Mutational analysis indicated wild-type activity for the Asp31Asn carboxylase. In contrast, the respective Trp157Arg and Thr591Lys activities were 8% and 0% that of wild-type carboxylase, and their compound heterozygosity can therefore account for functional VKD factor deficiency. The implications for carboxylase mechanism are discussed. PMID- 16720841 TI - Food for thought: honeybee foraging, memory, and acetylcholine. AB - Recent research implicates acetylcholine signaling through muscarinic receptors in structural changes that take place in the honeybee brain in response to foraging. These new findings are consistent with research from earlier studies implicating cholinergic signaling in associative learning as well as in the response to an enriched environment in mammals, which suggests that cholinergic signaling may play a critical role in learning and memory mechanisms across phyla. PMID- 16720842 TI - Fertility preservation in young women undergoing breast cancer therapy. AB - Breast cancer accounts for one third of all neoplasms seen in reproductive-age women and affects tens of thousands of women each year in that age group. The adjuvant chemotherapy regimens used for the treatment commonly affect fertility and cause premature ovarian failure. There have been recent advances in the field of fertility preservation, which can allow many of these breast cancer survivors to have children in the future. The most established option is embryo cryopreservation; oocyte cryopreservation can be considered in single women. Both of these approaches require approximately 2 weeks of ovarian stimulation beginning with the onset of the patient's menstrual cycle. Thus, it is crucial that these patients are referred to appropriate assisted reproduction centers as soon as they are diagnosed with breast cancer. Recently developed ovarian stimulation protocols using tamoxifen and letrozole can be used to increase the margin of safety in these patients. When and if a breast cancer patient does not have time to undergo ovarian stimulation prior to chemotherapy, ovarian cryopreservation for future autotransplantation can be offered as the last resort. The benefit of ovarian protection by gonadotropin-releasing hormone analogues is unproven and unlikely, and thus this treatment should not be offered as the sole method of fertility preservation. PMID- 16720843 TI - Benign breast diseases: classification, diagnosis, and management. AB - Benign breast diseases constitute a heterogeneous group of lesions including developmental abnormalities, inflammatory lesions, epithelial and stromal proliferations, and neoplasms. In this review, common benign lesions are summarized and their relationship to the development of subsequent breast cancer is emphasized. PMID- 16720844 TI - Introduction to the new section. When clinical trials are not an option. PMID- 16720845 TI - Therapeutic vaccines for prostate cancer. AB - Prostate cancer is the most common, noncutaneous cancer for men in the U.S., leading to more than 30,000 deaths a year. Vaccines for prostate cancer, which for several years have been shown to generate immunologic responses, are beginning to show significant clinical promise. At present, numerous therapeutic options are being investigated, including autologous and allogeneic whole-tumor cell vaccines, dendritic cell vaccines, and poxvirus-based vaccines. Advances in basic immunology have translated into new, more complex therapeutic strategies. The findings from current trials and the demonstrated potential to combine vaccines with conventional therapies herald a promising future for the treatment of prostate cancer. This review highlights recent advances and clinical trials in immunotherapy for prostate cancer, along with current thoughts on immunologic and clinical monitoring of these trials. PMID- 16720846 TI - Preventing chemotherapy errors. AB - A large amount of information on chemotherapy error prevention is available to the practicing oncologist. However, few oncologists have the time and resources to obtain the information and evaluate the evidence. Further, much of the information is generic and does not provide specific direction on how the information can be applied in clinical practice. This manuscript reviews principles of safe chemotherapy administration, identifies key actions to prevent chemotherapy errors, and provides suggestions on how the information can be incorporated into daily practice. PMID- 16720847 TI - RADPLAT: an alternative to surgery? AB - Head and neck cancer frequently presents at a late stage, leading to a poor prognosis despite optimal treatment with surgery and/or radiotherapy. Chemotherapy for advanced disease has shown little benefit as a single-modality treatment, and the use of concurrent chemoradiation is limited by problems with severe toxicity at higher doses. RADPLAT is the acronym used to describe a new technique, combining intra-arterial delivery of cisplatin with systemic neutralization by i.v. sodium thiosulphate, and concurrent radiotherapy. This allows very high cisplatin dose intensities to be used while potentially minimizing adverse systemic effects. Initial results suggest that excellent locoregional control rates are achievable in patients with unresectable disease, with a favorable side-effect profile when compared with conventional chemoradiation protocols. In addition, RADPLAT may potentially be of benefit in selected patients with resectable disease, allowing for preservation of organ function and quality of life without compromising locoregional control or survival. While current phase II data are encouraging, phase III randomized controlled trials are required in order to directly compare RADPLAT with i.v. chemoradiation therapy, the current standard of care. This article reviews the evolution of the RADPLAT concept, from initial clinical trials to its current application in the treatment of patients with advanced head and neck cancer. PMID- 16720848 TI - Lung cancer screening. AB - Advances in imaging technology have ushered in a new era for lung cancer screening in high-risk individuals using computed tomographic (CT) scans. Although most published studies are nonrandomized observational cohorts of volunteers, the ability of CT scans to detect early stage lung cancer is undisputable. What is unresolved is the ability of spiral CT screening to affect lung cancer-related mortality. A large randomized trial sponsored by the National Cancer Institute to address this question is currently under way. Genomic and proteomic approaches promise to complement the ability of spiral CT to detect early lung cancer in the next few years. Currently, the decision to screen for lung cancer should involve a careful discussion with the individuals involved about the potential advantages, costs, and drawbacks of the approach. PMID- 16720849 TI - Primary mediastinal large B-cell lymphoma. AB - Primary mediastinal large B-cell lymphoma represents a distinct entity with unique clinicopathologic features and a molecular gene-expression signature reminiscent of nodular sclerosis subtype of classical Hodgkin's lymphoma. Recent studies, including those using a refined molecular signature, suggest that the outcome is more favorable than that of diffuse large B-cell lymphoma. Using historical comparisons, dose-dense and dose-intensive regimens may be more effective than cyclophosphamide, doxorubicin, vincristine, and prednisone chemotherapy; however, the impact of adding rituximab to these regimens and effect on outcome comparisons is unknown. Clinical trials exploring these questions in addition to the benefit of consolidative radiotherapy are necessary to definitively answer these questions. PMID- 16720850 TI - Intravascular lymphoma: the oncologist's "great imitator". AB - Intravascular lymphoma (IVL) is a rare subtype of extranodal diffuse large B-cell lymphoma with a distinct presentation. Anatomically the disease is characterized by the proliferation of clonal lymphocytes within small vessels with relative sparing of the surrounding tissue. The clinical symptoms of the disease are dependent on the specific organ involvement, which most often includes the central nervous system and skin. Because of the various modes of presentation and the rarity of IVL, the diagnosis is often made postmortem. The diagnosis is almost exclusively made by surgical biopsy of a suspected site of involvement. Advances in imaging and immunohistochemistry have led to increasing antemortem diagnosis of this lymphoma. Although some patients with this disease may be curable with aggressive therapy, further research into novel treatment strategies is needed to improve outcome. Some potential insights into future therapies may be drawn from the small amount of basic science literature relevant to this entity. This review provides a concise, up-to-date summary of IVL. PMID- 16720851 TI - Ewing's sarcoma family of tumors: current management. AB - Ewing's sarcoma is the second most frequent primary bone cancer, with approximately 225 new cases diagnosed each year in patients less than 20 years of age in North America. It is one of the pediatric small round blue cell tumors, characterized by strong membrane expression of CD99 in a chain-mail pattern and negativity for lymphoid (CD45), rhabdomyosarcoma (myogenin, desmin, actin) and neuroblastoma (neurofilament protein) markers. Pathognomonic translocations involving the ews gene on chromosome 22 and an ets-type gene, most commonly the fli1 gene on chromosome 11, are implicated in the great majority of cases. Clinical presentation is usually dominated by local bone pain and a mass. Imaging reveals a technetium pyrophosphate avid lesion that, on plain radiograph, is destructive, diaphyseal and classically causes layered periosteal calcification. Magnetic resonance best defines the extent of the lesion. Biopsy should be undertaken by an experienced orthopedic oncologist. Approximately three quarters of patients have initially localized disease. About two thirds survive disease free. Management, preferably at a specialist center with a multi-disciplinary team, includes both local control-either surgery, radiation or a combination-and systemic chemotherapy. Chemotherapy includes cyclic combinations, incorporating vincristine, doxorubicin, cyclophosphamide, etoposide, ifosfamide and occasionally actinomycin D. Topotecan in combination with cyclophosphamide has shown preliminary activity. Patients with initially metastatic disease fare less well, with about one quarter surviving. Studies incorporating intensive therapy followed by stem cell infusion show no clear benefit. New approaches include anti angiogenic therapy, particularly since vascular endothelial growth factor is an apparent downstream target of the ews-fli1 oncogene. PMID- 16720852 TI - Teams: communication in multidisciplinary care. AB - Shortly before his death in 1995, Kenneth B. Schwartz, a cancer patient at Massachusetts General Hospital (MGH), founded The Kenneth B. Schwartz Center at MGH. The Schwartz Center is a nonprofit organization dedicated to supporting and advancing compassionate health care delivery that provides hope to the patient and support to caregivers, and encourages the healing process. The Center sponsors the Schwartz Center Rounds, a monthly multidisciplinary forum where caregivers reflect on important psychosocial issues faced by patients, their families, and their caregivers, and gain insight and support from fellow staff members. The evolving field of oncology increasingly requires a team of medical specialists working in unison to deliver optimal medical care. While this coordination may maximize the technical synergy of care, it can challenge interprofessional and interdisciplinary connections. Poor and miscommunication and conflicts between staff and between the family and providers adversely affect patient care and quality of life. Furthermore, lack of communication leaves a vacuum that sucks in fear. A recent Newsweek article highlighted the challenges of practicing in the age of high-tech medicine. The author had to beg for a prognosis for her critically ill and dying husband, with unhelpful subspecialists failing to communicate the bigger picture. This article explores the tough issue of how teams handle uncertainty and bad news and how patients and families can be better supported in the multifaceted paradigm of modern care. PMID- 16720854 TI - Topotecan continuous infusion: CA-125 responses including patients pretreated with other schedules of topotecan. PMID- 16720857 TI - Trastuzumab combined with paclitaxel after doxorubicin and cyclophosphamide for operable HER2-positive breast cancer. PMID- 16720859 TI - Regarding "randomized comparison of epoetin alfa and darbepoetin alfa in anemic patients with cancer receiving chemotherapy". PMID- 16720861 TI - Response to "helping melanoma patients decide whether to choose adjuvant high dose interferon-alpha2b". PMID- 16720863 TI - Disinhibitory pathways for control of sodium transport: regulation of ENaC by SGK1 and GILZ. AB - Regulation of ENaC occurs at several levels. The principal hormonal regulator of ENaC, aldosterone, acts through the mineralocorticoid receptor to modulate ENaC mediated sodium transport, and considerable attention has focused on defining the components of the early phase of this response. Two genes, SGK1 and GILZ, have now been implicated in this regulation. While the functional significance of SGK1 in mediating aldosterone effects is well established, new evidence has enhanced our understanding of the mechanisms of SGK1 action. In addition, recent work demonstrates a novel role for GILZ in the stimulation of ENaC-mediated sodium transport. Interestingly, both SGK1 and GILZ appear to negatively regulate tonic inhibition of ENaC and thus use disinhibition to propagate the rapid effects of aldosterone to increase sodium reabsorption in tight epithelia. PMID- 16720864 TI - Transforming growth factor-beta1 differentially mediates fibronectin and inflammatory cytokine expression in kidney tubular cells. AB - Transforming growth factor-beta(1) (TGF-beta(1)) is not only an important fibrogenic but also immunomodulatory cytokine in the human kidney. We have recently demonstrated that TGF-beta(1) induces interleukin-8 (IL-8), macrophage chemoattractant protein-1 (MCP-1), and fibronectin production in renal proximal tubular (HK-2) cells. However, the unique dependence of IL-8, MCP-1, and fibronectin on TGF-beta(1) expression is unknown. The TGF-beta(1) gene was effectively silenced in HK-2 cells using small-interference (si) RNA. Basal secretion of IL-8 and MCP-1 decreased (both P < 0.05) but, paradoxically, fibronectin increased (P < 0.05) in TGF-beta(1)-silenced cells compared with cells transfected with nonspecific siRNA. Significant increases were observed in mRNA for the TGF-beta(2) (P < 0.05), TGF-beta(3) (P < 0.05) isoforms and pSmad2 (P < 0.05), which were reflected in protein expression. Concurrent exposure to pan-specific TGF-beta antibody reversed the observed increase in fibronectin expression, suggesting that TGF-beta(2) and TGF-beta(3) isoforms mediate the increased fibronectin expression in TGF-beta(1)-silenced cells. An increase in the DNA binding activity of activator protein-1 (AP-1; P < 0.05) was also observed in TGF-beta(1)-silenced cells. In contrast, nuclear factor-kappaB (NF kappaB) DNA binding activity was significantly decreased (P < 0.0005). These studies demonstrate that TGF-beta(1) is a key regulator of IL-8 and MCP-1, whereas fibronectin expression is regulated by a complex interaction between the TGF-beta isoforms in the HK-2 proximal tubular cell line. Decreased expression of TGF-beta(1) reduces chemokine production in association with reduced NF-kappaB DNA binding activity, suggesting that immunomodulatory pathways in the kidney are specifically dependent on TGF-beta(1). Conversely, decreased expression of TGF beta(1) results in increased TGF-beta(2), TGF-beta(3), AP-1, and pSmad2 that potentially mediates the observed increase in fibronectin. PMID- 16720865 TI - Adverse events after unenhanced and monomeric and dimeric contrast-enhanced CT: a prospective randomized controlled trial. AB - PURPOSE: To prospectively evaluate adverse events (AEs) in patients who underwent nonionic monomeric or dimeric contrast media-enhanced computed tomography (CT) and to compare these effects with AEs in patients who underwent unenhanced CT. MATERIALS AND METHODS: Local ethics committee approval and written informed consent were obtained. Patients were randomly assigned to the dimeric group, monomeric group, or concurrent control group. Acute (occurring during or within 1 hour after contrast agent injection) and delayed (occurring between 1 hour and 7 days after contrast agent injection) AEs were evaluated and categorized (mild, not requiring treatment; moderate, self-limiting AE requiring simple treatment; severe, AE requiring extensive treatment or endangering life). The two-sided Fisher exact test and the Mann-Whitney U and Wilcoxon signed rank tests were used for statistical analysis. RESULTS: A total of 895 patients were recruited. Acute AEs were noted significantly (P < .05) more often in the monomeric group (44.8% [133 of 297 patients]) than in the dimeric (23.7% [71 of 300 patients]) or control (9.4% [28 of 298 patients]) groups. Two moderate acute AEs were noted in one patient in each contrast agent group; all other acute AEs were mild. There was no significant difference in the overall incidence of delayed AEs between the dimeric and monomeric groups (53.1% [139 of 262 patients] vs 50.8% [131 of 258 patients]). Delayed cutaneous AEs were noted significantly (P < .05) more often in the dimeric group. A total 16 AEs (2.0%) were moderate; no AE was severe. CONCLUSION: The dimeric contrast agent caused fewer acute AEs than the monomeric contrast agent; however, the dimeric and monomeric agents caused a similar overall number of delayed AEs. Delayed cutaneous symptoms were noted more often with the dimeric contrast agent. Both contrast agents were safe in that no severe AEs and only a few moderate AEs were observed. PMID- 16720866 TI - Augmented reality visualization for CT-guided interventions: system description, feasibility, and initial evaluation in an abdominal phantom. AB - The purpose of this study was to evaluate the feasibility and performance of an augmented reality (AR) visualization prototype for virtual computed tomography (CT)-guided interventional procedures in a multimodality abdominal phantom. With the aid of AR guidance, three radiologists performed 30 attempts at targeting simulated liver lesions of different sizes (range, 5-15 mm) with a biopsy needle. The position of the needle tip relative to the lesion was verified by using ultrasonography and CT. With AR guidance, lesions were successfully targeted with the first needle pass in all cases. On the basis of these results, AR visualization for CT-guided intervention appears feasible and allows intuitive and accurate lesion targeting in a phantom. SUPPLEMENTAL MATERIAL: radiology.rsnajnls.org/cgi/content/full/2401040018/DC1 PMID- 16720867 TI - Hepatic vein transit time of SonoVue: a comparative study with Levovist. AB - PURPOSE: To prospectively compare transit times of Levovist and SonoVue in healthy volunteers and patients with biopsy-proved hepatitis C-related liver disease. MATERIALS AND METHODS: Institutional review board approval and informed consent were obtained. Forty patients and 25 healthy volunteers were examined. Subjects fasted, a bolus of SonoVue (0.6 mL) was injected into a cubital fossa vein, and hepatic venous time-intensity profiles were measured with spectral Doppler tracing. This was repeated with two injections of Levovist (2 g) and another injection of SonoVue. Time-intensity curves of spectral Doppler signals of right and middle hepatic veins were analyzed. A sustained signal intensity increase of 10% above baseline levels indicated hepatic vein transit time (HVTT). Carotid artery audio intensity was measured in volunteers. Analysis of variance and t tests were used for statistical analysis. RESULTS: Twelve patients had mild hepatitis; 18, moderate or severe hepatitis; and 10, cirrhosis. Mean HVTTs in control, mild hepatitis, moderate or severe hepatitis, and cirrhosis groups were 38.3 seconds +/- 2.4 (standard error), 47.5 seconds +/- 6.5, 29.5 seconds +/- 10.8, and 17.6 seconds +/- 5.0, respectively, with Levovist (P < .001) and 29.4 seconds +/- 6.9, 27.4 seconds +/- 9.3, 22.9 seconds +/- 4.7, and 16.4 seconds +/- 4.9, respectively, with SonoVue (P < .001). HVTT decreased as severity increased at imaging with both contrast agents. There was no significant difference in HVTT between mild and moderate hepatitis groups with SonoVue; however, there were significant differences in HVTT between all patient groups with Levovist. HVTT of SonoVue was shorter than that of Levovist in all groups (P < .001) except the cirrhosis group; in this group, HVTT of the two contrast agents was similar (P = .05). No difference was observed in mean cardiopulmonary transit time for SonoVue or Levovist (9.1 seconds +/- 2.4 [standard error] and 8.4 seconds +/- 2.5, respectively, P = .18). CONCLUSION: HVTT was significantly shorter with SonoVue than with Levovist; there was no significant difference in cardiopulmonary transit time. PMID- 16720868 TI - Transrenal fixation of aortic stent-grafts: short- to midterm effects on renal function--a systematic review. AB - PURPOSE: To perform a systematic review of the short- to midterm effects of transrenal fixation of aortic stent-grafts on renal function in patients with abdominal aortic aneurysms. MATERIALS AND METHODS: A search of the PubMed, MEDLINE, and EMBASE databases for English-language literature was performed. Studies with at least 10 patients were included for data analysis. Only studies on transrenal fixation of aortic stent-grafts that included follow-up results for renal function were included. A log-linear model was used for meta-analysis to compare transrenal fixation with infrarenal fixation. RESULTS: Twenty-two studies met the inclusion criteria. Because two studies analyzed the same group of patients, one was excluded, for a total of 21 studies. Comparisons between transrenal fixation and infrarenal fixation were found in seven studies. For transrenal versus infrarenal fixation, the combined odds ratio, 95% confidence interval, and P value were found to be statistically significant with respect to postprocedural renal infarction only (combined odds ratio, 5.189; 95% confidence interval: 3.198, 8.420; P < .001). No significant difference was found between transrenal and infrarenal fixation with respect to renal dysfunction, renal artery occlusion, or endoleaks (P > .05). CONCLUSION: Transrenal fixation of aortic stent-grafts seems to be a relatively safe alternative compared with infrarenal fixation in terms of short- to midterm follow-up. Postprocedural renal infarction, however, was significantly higher for transrenal fixation. PMID- 16720869 TI - Arterial calcifications seen on mammograms: cardiovascular risk factors, pregnancy, and lactation. AB - PURPOSE: To retrospectively assess if mammographic calcium deposits are related to coronary heart disease (CHD) risk factors and reproductive factors in a subset of women participating in the European Prospective Investigation into Cancer and Nutrition study. MATERIALS AND METHODS: The study was approved by the institutional review board of the University Medical Center Utrecht; informed consent was obtained. Mammograms were evaluated by two radiologists for the presence of breast arterial calcifications (BAC) in the Prospect cohort, a breast cancer screening population of women aged 49-70 years (mean, 57 years) within the European Prospective Investigation into Cancer and Nutrition study. Cardiovascular risk factors and reproductive factors were examined for independent effects on the prevalence of BAC. Logistic regression analysis was performed. RESULTS: BAC was present in 194 of 1699 (11%) women and increased with age to 20% in the highest quartile of age (mean, 66 years). The odds ratio was 4.7 in the highest versus the lowest quartile of age (95% confidence interval [CI], 2.9, 7.6). After adjustment for age, no significant association was found between BAC and traditional cardiovascular risk factors. Current smoking was inversely related to BAC (odds ratio, 0.6; 95% CI: 0.4, 0.9). BAC was prevalent in 2.5% of nulliparous women, in 9% of women with one or two children, and in 17% of women with three or more children (odds ratio, 7.2; 95% CI: 2.9, 18.0). Breast feeding after pregnancy was significantly associated with BAC in women who ever were pregnant (odds ratio, 2.2; 95% CI: 1.4, 3.6). CONCLUSION: Calcifications in arteries on mammograms are associated with increasing age, pregnancy, and lactation but not with various cardiovascular risk factors. PMID- 16720870 TI - Does multi-detector row CT pulmonary angiography reduce the incremental value of indirect CT venography compared with single-detector row CT pulmonary angiography? AB - PURPOSE: To compare retrospectively the incremental value of indirect computed tomographic (CT) venography performed after multi-detector row CT pulmonary angiography and single-detector row CT pulmonary angiography for the diagnosis of venous thromboembolism (VTE). MATERIALS AND METHODS: The institutional ethics committee approved this study; informed consent was not required. The authors retrospectively reviewed results of 1100 combined single-detector row CT pulmonary angiographic and indirect CT venographic examinations (542 men, 558 women; mean age, 61 years +/- 17 [standard deviation]) (group 1) and 308 combined multi-detector row CT pulmonary angiographic and indirect CT venographic examinations (150 men, 158 women; mean age, 62 years +/- 18) (group 2), performed in 1408 patients suspected of having pulmonary embolism (PE). Frequency of deep venous thrombosis (DVT), PE, and VTE, and the incremental value of indirect CT venography were recorded in both groups. Data were compared by means of the Student t test for continuous data and z statistics for independent proportions. RESULTS: VTE, PE, and DVT were found in 23.3% (n = 256), 19.9% (n = 219), and 18.3% (n = 201) of the 1100 patients in group 1, respectively, and in 23.7% (n = 73), 17.2% (n = 53), and 18.8% (n = 58) of the 308 patients in group 2, respectively (P values ranging from .273 to .876). The incremental value of indirect CT venography was 14.4% (37 of 256 patients) in group 1 and 27.4% (20 of 73 patients) in group 2. CONCLUSION: Despite potential improved accuracy of multi detector row CT pulmonary angiography for the diagnosis of PE, the addition of indirect CT venography increased the diagnosis of VTE in 27.4% of patients. PMID- 16720871 TI - Evaluation of living renal donors: accuracy of three-dimensional 16-section CT. AB - PURPOSE: To retrospectively assess the sensitivity and specificity of three dimensional (3D) 16-section computed tomography (CT) in the evaluation of vessels, pelvicalyceal system, and ureters in living renal donors, with surgical findings as the reference standard. MATERIALS AND METHODS: This was a HIPAA compliant study. Institutional review board approval was obtained for the review of subjects' medical records and data analysis, with waiver of informed consent. Forty-six renal donors (18 men, 28 women; mean age, 42 years) were examined with 16-section CT. Two blinded reviewers independently studied renal vascular and urographic anatomy of each donor CT scans by fist using 3D images alone, then transverse images alone, and finally transverse and 3D data set. Image quality, degree of diagnostic confidence, and time used for review were recorded. Sensitivity and specificity were calculated. RESULTS: For 3D images, transverse images, and transverse in conjunction with 3D data sets, the respective sensitivity and specificity of CT in evaluation of accessory arteries by reviewer 1 were 100% and 100%, 89% and 100%, and 100% and 100%, and those by reviewer 2 were 89% and 97%, 89% and 100%, and 89% and 100%; the respective sensitivity and specificity in evaluation of venous anomalies by reviewer 1 were 100% and 98%, 100% and 98%, and 100% and 98%, and those by reviewer 2 were 100% and 98%, 100% and 95%, and 100% and 98%. For focused comprehensive assessment of renal donors with 3D scans alone, a reviewer on average (average of reviewers 1 and 2) used 2.4 minutes per scan, demonstrated full confidence in 93%, and rated the quality as excellent in 76%. CONCLUSION: For focused assessment of renal vascular and urographic anatomy, review of 3D data set alone provides high sensitivity and specificity with regard to findings seen at surgery. PMID- 16720872 TI - Improved tumor destruction with arsenic trioxide and radiofrequency ablation in three animal models. AB - PURPOSE: To assess the extent of tumor blood flow reduction that is achievable with arsenic trioxide (As2O3) and the effect of As2O3 on radiofrequency (RF) induced coagulation. MATERIALS AND METHODS: All animal protocols and experiments were approved by an institutional animal care and use committee before the start of the study. Experiments were conducted in three tumor models: intrarenal VX2 sarcoma in 27 rabbits, RCC 786-0 human renal cell carcinoma in 24 nude mice, and R3230 mammary adenocarcinoma in 40 rats. One dose (0-7.5 mg per kilogram of body weight) of As2O3 was administered (intraperitoneally in rodents, intravenously in rabbits) 1, 6, or 24 hours before standardized RF ablation, which was performed by using a 1-cm active tip, with mean temperatures of 70 degrees C +/- 2 (standard deviation) for 5 minutes in rodents and 90 degrees C +/- 2 for 6 minutes in rabbits. Laser Doppler flowmetry was used to quantify changes in blood flow, which were compared with diameters of induced tumor coagulation. Comparisons between groups were performed by using Student t tests or analysis of variance. The strengths of correlations between As2O3, tumor blood flow, and RF induced coagulation were assessed by using linear and higher-order regression models and reported as R2 computations. RESULTS: Administration of As2O3 significantly (P < .05) reduced blood flow and increased tumor destruction in all tumor models. In VX2 sarcoma tumors, 1 mg/kg As2O3 reduced mean tumor blood flow to 46% +/- 13 of the normal value. The mean resultant coagulation (1.1 cm +/- 0.1) was significantly greater than that achieved with RF ablation alone (0.6 cm +/- 0.1, P < .01). In RCC 786-0 and R3230 tumors, 5 mg/kg As2O3 reduced mean tumor blood flow to 57% +/- 6 and 46% +/- 6 of normal, respectively, increasing mean ablation extent to 0.8 cm +/- 0.1 for both models, compared with those achieved with the control treatment (0.6 cm +/- 0.1 and 0.5 cm +/- 0.1, respectively; P < .05 for both comparisons). Dose studies revealed correlations between drug dose, tumor blood flow, and RF-induced coagulation in all three tumor models (R2 = 0.60-0.79). Maximal RF synergy was observed 1 hour after As2O3 administration. CONCLUSION: As2O3 administration represents a transient noninvasive method of reducing tumor blood flow during RF ablation, enabling larger zones of tumor destruction in multiple tumor models. PMID- 16720873 TI - Getting to the guts of enteric nervous system development. AB - Scientists from around the world gathered in New York City recently to discuss the latest research on enteric nervous system development at a meeting organised by Alan Burns and Heather Young. The participants enjoyed 3 days of presentations that spurred active conversations and highlighted the rapidly advancing research in this field. PMID- 16720874 TI - Mechanisms of elongation in embryogenesis. AB - Here, I discuss selected examples of elongation in embryogenesis to identify common and unique mechanisms, useful questions for further work, and new systems that offer opportunities for answering these questions. Fiber-wound, hydraulic mechanisms of elongation highlight the importance of biomechanical linkages of otherwise unrelated cellular behaviors during elongation. Little-studied examples of elongation by cell intercalation offer opportunities to study new aspects of this mode of elongation. Elongation by oriented cell division highlights the problem of mitotic spindle orientation and the maintenance of cell-packing patterns in anisotropic force environments. The balance of internal cell-adhesion and external traction forces emerges as a key issue in the formation of elongate structures from compact ones by directed migration. PMID- 16720875 TI - Gli3-mediated somitic Fgf10 expression gradients are required for the induction and patterning of mammary epithelium along the embryonic axes. AB - Little is known about the regulation of cell fate decisions that lead to the formation of five pairs of mammary placodes in the surface ectoderm of the mouse embryo. We have previously shown that fibroblast growth factor 10 (FGF10) is required for the formation of mammary placodes 1, 2, 3 and 5. Here, we have found that Fgf10 is expressed only in the somites underlying placodes 2 and 3, in gradients across and within these somites. To test whether somitic FGF10 is required for the formation of these two placodes, we analyzed a number of mutants with different perturbations of somitic Fgf10 gradients for the presence of WNT signals and ectodermal multilayering, markers for mammary line and placode formation. The mammary line is displaced dorsally, and formation of placode 3 is impaired in Pax3ILZ/ILZ mutants, which do not form ventral somitic buds. Mammary line formation is impaired and placode 3 is absent in Gli3Xt-J/Xt-J and hypomorphic Fgf10 mutants, in which the somitic Fgf10 gradient is shortened dorsally and less overall Fgf10 is expressed, respectively. Recombinant FGF10 rescued mammogenesis in Fgf10(-/-) and Gli3Xt-J/Xt-J flanks. We correlate increasing levels of somitic FGF10 with progressive maturation of the surface ectoderm, and show that full expression of somitic Fgf10, co-regulated by GLI3, is required for the anteroposterior pattern in which the flank ectoderm acquires a mammary epithelial identity. We propose that the intra-somitic Fgf10 gradient, together with ventral elongation of the somites, determines the correct dorsoventral position of mammary epithelium along the flank. PMID- 16720876 TI - Axis specification in the spider embryo: dpp is required for radial-to-axial symmetry transformation and sog for ventral patterning. AB - The mechanism by which Decapentaplegic (Dpp) and its antagonist Short gastrulation (Sog) specify the dorsoventral pattern in Drosophila embryos has been proposed to have a common origin with the mechanism that organizes the body axis in the vertebrate embryo. However, Drosophila Sog makes only minor contributions to the development of ventral structures that hypothetically correspond to the vertebrate dorsum where the axial notochord forms. In this study, we isolated a homologue of the Drosophila sog gene in the spider Achaearanea tepidariorum, and characterized its expression and function. Expression of sog mRNA initially appeared in a radially symmetrical pattern and later became confined to the ventral midline area, which runs axially through the germ band. RNA interference-mediated depletion of the spider sog gene led to a nearly complete loss of ventral structures, including the axial ventral midline and the central nervous system. This defect appeared to be the consequence of dorsalization of the ventral region of the germ band. By contrast, the extra embryonic area formed normally. Furthermore, we showed that embryos depleted for a spider homologue of dpp failed to break the radial symmetry, displaying evenly high levels of sog expression except in the posterior terminal area. These results suggest that dpp is required for radial-to-axial symmetry transformation of the spider embryo and sog is required for ventral patterning. We propose that the mechanism of spider ventral specification largely differs from that of the fly. Interestingly, ventral specification in the spider is similar to the process in vertebrates in which the antagonism of Dpp/BMP signaling plays a central role in dorsal specification. PMID- 16720877 TI - Drosophila talin and integrin genes are required for maintenance of tracheal terminal branches and luminal organization. AB - Epithelial tubes that compose many organs are typically long lasting, except under specific developmental and physiological conditions when network remodeling occurs. Although there has been progress elucidating mechanisms of tube formation, little is known of the mechanisms that maintain tubes and destabilize them during network remodeling. Here, we describe Drosophila tendrils mutations that compromise maintenance of tracheal terminal branches, fine gauge tubes formed by tracheal terminal cells that ramify on and adhere tightly to tissues in order to supply them with oxygen. Homozygous tendrils terminal cell clones have fewer terminal branches than normal but individual branches contain multiple convoluted lumens. The phenotype arises late in development: terminal branches bud and form lumens normally early in development, but during larval life lumens become convoluted and mature branches degenerate. Their lumens, however, are retained in the remaining branches, resulting in the distinctive multi-lumen phenotype. Mapping and molecular studies demonstrate that tendrils is allelic to rhea, which encodes Drosophila talin, a large cytoskeletal protein that links integrins to the cytoskeleton. Terminal cells mutant for myospheroid, the major Drosophila beta-integrin, or doubly mutant for multiple edematous wings and inflated alpha-integrins, also show the tendrils phenotype, and localization of myospheroid beta-integrin protein is disrupted in tendrils mutant terminal cells. The results provide evidence that integrin-talin adhesion complexes are necessary to maintain tracheal terminal branches and luminal organization. Similar complexes may stabilize other tubular networks and may be targeted for inactivation during network remodeling events. PMID- 16720878 TI - Testing hypotheses for the functions of APC family proteins using null and truncation alleles in Drosophila. AB - Adenomatous polyposis coli (APC) is mutated in colon cancers. During normal development, APC proteins are essential negative regulators of Wnt signaling and have cytoskeletal functions. Many functions have been proposed for APC proteins, but these have often rested on dominant-negative or partial loss-of-function approaches. Thus, despite intense interest in APC, significant questions remain about its full range of cellular functions and about how mutations in the gene affect these. We isolated six new alleles of Drosophila APC2. Two resemble the truncation alleles found in human tumors and one is a protein null. We generated ovaries and embryos null for both APC2 and APC1, and assessed the consequences of total loss of APC function, allowing us to test several previous hypotheses. Surprisingly, although complete loss of APC1 and APC2 resulted in strong activation of Wingless signaling, it did not substantially alter cell viability, cadherin-based adhesion, spindle morphology, orientation or selection of division plane, as predicted from previous studies. We also tested the hypothesis that truncated APC proteins found in tumors are dominant negative. Two mutant proteins have dominant effects on cytoskeletal regulation, affecting Wnt-independent nuclear retention in syncytial embryos. However, they do not have dominant negative effects on Wnt signaling. PMID- 16720881 TI - Learning to love big brother. PMID- 16720880 TI - Fgf8 is required for anterior heart field development. AB - In the mouse embryo, the splanchnic mesodermal cells of the anterior heart field (AHF) migrate from the pharynx to contribute to the early myocardium of the outflow tract (OT) and right ventricle (RV). Recent studies have attempted to distinguish the AHF from other precardiac populations, and to determine the genetic and molecular mechanisms that regulate its development. Here, we have used an Fgf8lacZ allele to demonstrate that Fgf8 is expressed within the developing AHF. In addition, we use both a hypomorphic Fgf8 allele (Fgf8neo) and Cre-mediated gene ablation to show that Fgf8 is essential for the survival and proliferation of the AHF. Nkx2.5Cre is expressed in the AHF, primary heart tube and pharyngeal endoderm, while TnT-Cre is expressed only within the specified heart tube myocardium. Deletion of Fgf8 by Nkx2.5Cre results in a significant loss of the Nkx2.5Cre lineage and severe OT and RV truncations by E9.5, while the remaining heart chambers (left ventricle and atria) are grossly normal. These defects result from significant decreases in cell proliferation and aberrant cell death in both the pharyngeal endoderm and splanchnic mesoderm. By contrast, ablation of Fgf8 in the TnT-Cre domain does not result in OT or RV defects, providing strong evidence that Fgf8 expression is crucial in the pharyngeal endoderm and/or overlying splanchnic mesoderm of the AHF at a stage prior to heart tube elongation. Analysis of downstream signaling components, such as phosphorylated-Erk and Pea3, identifies the AHF splanchnic mesoderm itself as a target for Fgf8 signaling. PMID- 16720879 TI - Required, tissue-specific roles for Fgf8 in outflow tract formation and remodeling. AB - Fibroblast growth factor 8 (Fgf8) is a secreted signaling protein expressed in numerous temporospatial domains that are potentially relevant to cardiovascular development. However, the pathogenesis of complex cardiac and outflow tract defects observed in Fgf8-deficient mice, and the specific source(s) of Fgf8 required for outflow tract formation and subsequent remodeling are unknown. A detailed examination of the timing and location of Fgf8 production revealed previously unappreciated expression in a subset of primary heart field cells; Fgf8 is also expressed throughout the anterior heart field (AHF) mesoderm and in pharyngeal endoderm at the crescent and early somite stages. We used conditional mutagenesis to examine the requirements for Fgf8 function in these different expression domains during heart and outflow tract morphogenesis. Formation of the primary heart tube and the addition of right ventricular and outflow tract myocardium depend on autocrine Fgf8 signaling in cardiac crescent mesoderm. Loss of Fgf8 in this domain resulted in decreased expression of the Fgf8 target gene Erm, and aberrant production of Isl1 and its target Mef2c in the anterior heart field, thus linking Fgf8 signaling with transcription factor networks that regulate survival and proliferation of the anterior heart field. We further found that mesodermal- and endodermal-derived Fgf8 perform specific functions during outflow tract remodeling: mesodermal Fgf8 is required for correct alignment of the outflow tract and ventricles, whereas activity of Fgf8 emanating from pharyngeal endoderm regulates outflow tract septation. These findings provide a novel insight into how the formation and remodeling of primary and anterior heart field-derived structures rely on Fgf8 signals from discrete temporospatial domains. PMID- 16720883 TI - Regulation of reproductive function in athletic women: an investigation of the roles of energy availability and body composition. PMID- 16720884 TI - The myth of the female athlete triad. PMID- 16720885 TI - Low back pain investigations and prognosis: a review. AB - Low back pain is reviewed in terms of when investigations are useful and its clinical course. Despite the extensive evaluation of the accuracy of investigations such as radiography, magnetic resonance imaging, and myelography, there is a surprising dearth of research to inform their use in primary care. There is no clear evidence on which to base judgments for selection of appropriate tests to confirm or exclude low back pain pathology. It appears that investigations are rarely necessary for low back pain. Specific investigations should be ordered to identify a particular pathology but should not be ordered routinely for general screening. In the absence of pathology, low back pain and its associated disability improve rapidly in the first weeks after onset, but, in contradiction to all guidelines, both commonly persist and the best evidence suggests that recurrences are common. PMID- 16720886 TI - The effects of creatine supplementation on selected factors of tennis specific training. AB - BACKGROUND: Creatine supplementation is popular among tennis players but it is not clear whether it actually enhances tennis performance. OBJECTIVES: To examine the effects of creatine supplementation on tennis specific performance indices. METHODS: In a randomised, double blind design, 36 competitive male tennis players (24 creatine, mean (SD) age, 22.5 (4.9) years; 12 placebo, 22.8 (4.8) years) were tested at baseline, after six days of creatine loading, and after a maintenance phase of four weeks (14 creatine, 10 placebo). Serving velocity (10 serves), forehand and backhand velocity (three series of 5x8 strokes), arm and leg strength (bench press and leg press), and intermittent running speed (three series of five 20 metre sprints) were measured. RESULTS: Compared with placebo, neither six days nor five weeks of creatine supplementation had a significant effect on serving velocity (creatine: +2 km/h; placebo: +2 km/h, p = 0.90); forehand velocity (creatine: +4 km/h; placebo: +4 km/h, p = 0.80), or backhand velocity (creatine: +3 km/h; placebo: +1 km/h, p = 0.38). There was also no significant effect of creatine supplementation on repetitive sprint power after 5, 10, and 20 metres, (creatine 20 m: -0.03 m/s; placebo 20 m: +0.01 m/s, p = 0.18), or in the strength of the upper and lower extremities. CONCLUSIONS: Creatine supplementation is not effective in improving selected factors of tennis specific performance and should not be recommended to tennis players. PMID- 16720887 TI - Stress fractures of the femoral shaft in athletes: a new treatment algorithm. AB - BACKGROUND: Femoral shaft stress fractures in athletes are not common but pose a great diagnostic challenge to clinicians. Because of few clinical signs, diagnosis and treatment are often delayed. Furthermore, if not treated correctly, these fractures are well known for complications and difficulties. OBJECTIVE: To develop a well structured and reproducible treatment algorithm for athletes with femoral shaft stress fractures. METHODS: The proposed algorithm is carried out in four phases, each lasting three weeks, and the move to the next phase is based on the result of the tests carried out at the end of the previous phase. Over nine years, we treated seven top level athletes, aged 17-21. In all athletes, diagnosis was based on physical examination, plain radiographs, and bone scan. RESULTS: As a result of the treatment method, all the athletes were fully engaged in athletic activity 12-18 weeks after the beginning of treatment. After completion of the treatment, the athletes were followed up for 48-96 months. During the follow up, there was no recurrence of discomfort or pain, and all the athletes eventually returned to competition level. CONCLUSION: These results and data available from the literature suggest that the algorithm is the optimal treatment protocol for femoral shaft stress fractures in athletes, avoiding the common complications and difficulties. PMID- 16720889 TI - Does history of concussion affect current cognitive status? AB - The association between self reported history of concussion and current neurocognitive status is controversial. Some football studies suggest that athletes with a history of concussion display cognitive impairment relative to athletes with no history of concussion, but other studies have not been able to reproduce such findings. This study shows that there is no relation between the number of previous self reported episodes of concussion and current cognitive state, directly contradicting the findings of previous research. PMID- 16720888 TI - Changes in exercise characteristics, maximal voluntary contraction, and explosive strength during prolonged tennis playing. AB - OBJECTIVES: To examine changes in exercise characteristics, maximal voluntary contraction, and explosive strength during prolonged tennis playing. METHODS: Maximal isometric voluntary contraction (MVC), leg stiffness (hopping), and peak power in squat (SJ) and countermovement (CMJ) jumps were measured before, every 30 minutes during, and 30 minutes after a three hour tennis match in 12 well trained players. Heart rate (HR), the effective playing time (EPT), rating of perceived exertion (RPE), and muscle soreness of knee extensors were also measured. RESULTS: Decreases in MVC (-9%; p < 0.05) and leg stiffness (-9%; p = 0.17) were observed after the match and were significantly correlated (r = 0.66; p = 0.05). Peak power in SJ and CMJ tests was maintained during the match but was lower (p < 0.001) 30 minutes after. Average HR and EPT were 144 (8) beats/min and 21 (4)% respectively. A strong correlation was found between EPT and HR (r = 0.93; p < 0.05). RPE and muscle soreness increased linearly during the exercise and were significantly correlated (r = 0.99; p < 0.001). CONCLUSIONS: Progressive reductions in maximal voluntary strength and leg stiffness highly correlated with increases in perceived exertion and muscle soreness were observed throughout a three hour tennis match, whereas explosive strength was maintained and decreased only after the match. These alterations may result in less efficient on-court movement and stroke production. They are, however, lower than those reported during continuous exercise of the same duration. The intermittent pattern of tennis and the numerous stretch-shortening cycle movements partly explain these results. PMID- 16720890 TI - Calf muscle wasting after tibial shaft fracture. AB - OBJECTIVES: To study the long term effect of tibial shaft fractures treated by immobilisation in a long leg cast on the calf muscle bulk. METHODS: Computed tomography scans were performed at fixed points on the lower legs of 23 non professional athletes who sustained closed tibial fractures 16 years previously. Length of immobilisation was determined from the hospital records. All the fractures were treated non-operatively. The cross sectional area of the various leg compartments was measured and compared with the non-injured leg. RESULTS: There was a significant reduction in cross sectional area of the posterior compartment (p<0.001, Student's t test). No such difference was seen in the anterolateral compartment. CONCLUSION: Tibial fractures treated non-operatively are associated with significant long term calf muscle wasting. PMID- 16720891 TI - Breathless in the bath. AB - The case is reported of a male track and field athlete with breathing difficulties at rest and during exercise, which were exacerbated in the supine position and during water immersion. Right hemidiaphragmatic paralysis was diagnosed. The cause of this relatively benign disorder is not known and there are no serious clinical implications. There is no treatment, but a continuation of exercise together with interventions to strengthen the subsidiary inspiratory muscles is recommended. PMID- 16720892 TI - Cognitive deterioration associated with an expedition in an extreme desert environment. AB - BACKGROUND: Prolonged exposure to extreme environments may result in cognitive changes that may influence decision making ability and increasing risk of injury or death. OBJECTIVE: To measure the cognitive performance of a healthy man as he completed a 17 day desert expedition. METHOD: A computer based cognitive test battery, subjective cognitive rating scale, and measures of physical characteristics were used. Objective cognitive performance was compared with the performance of eight age matched men who remained in their own homes. RESULTS: The speed of psychomotor, attentional, and executive functions decreased as the expedition progressed, but the accuracy of performance remained unaffected. Although some impairments were large, they resolved completely once the expedition was completed. Subjective ratings indicated that the subject had insight into his failing cognitive performance during the expedition. CONCLUSIONS: Cognitive performance can be measured repeatedly throughout an expedition in an extreme environment. Cognitive impairment may occur. PMID- 16720893 TI - Thematic review series: patient-oriented research. Nutritional determinants of insulin resistance. AB - Interpreting the literature relating to the nutritional determinants of insulin resistance is complicated by the wide range of methods used to determine insulin sensitivity. Excess adiposity is unquestionably the most important determinant of insulin resistance, although the effect may be tempered by a relatively high proportion of lean body mass. Weight loss is associated with improved insulin sensitivity. Thus, diet-related factors that promote excessive energy intake may be regarded as promoters of insulin resistance. In the context of energy balance, diets characterized by high intakes of saturated fat and low intakes of dietary fiber are associated with reduced insulin sensitivity. Total fat intakes greater than the usually consumed range appear to promote insulin resistance, although the relative proportions of total fat and carbohydrate within the usual range appear unimportant. Monounsaturated fatty acids with a cis configuration and fiber-rich carbohydrate foods appear to be appropriate substitutes for saturated fatty acids and rapidly digested glycemic carbohydrates. In animal studies, n-3 unsaturated fatty acids have been shown to enhance insulin sensitivity and fructose and sucrose to increase insulin resistance. However, human data are limited. Large prospective studies currently being conducted should confirm the most appropriate macronutrient composition of diets for preventing and treating insulin resistance as well as establishing whether a range of candidate genes explains the variation in response to dietary change. PMID- 16720894 TI - Thematic review series: patient-oriented research. What we have learned about VLDL and LDL metabolism from human kinetics studies. AB - Lipoprotein metabolism is the result of a complex network of many individual components. Abnormal lipoprotein concentrations can result from changes in the production, conversion, or catabolism of lipoprotein particles. Studies in hypolipoproteinemia and hyperlipoproteinemia have elucidated the processes that control VLDL secretion as well as VLDL and LDL catabolism. Here, we review the current knowledge regarding apolipoprotein B (apoB) metabolism, focusing on selected clinically relevant conditions. In hypobetalipoproteinemia attributable to truncations in apoB, the rate of secretion is closely linked to the length of apoB. On the other hand, in patients with the metabolic syndrome, it appears that substrate, in the form of free fatty acids, coupled to the state of insulin resistance can induce hypersecretion of VLDL-apoB. Studies in patients with familial hypercholesterolemia, familial defective apoB, and mutant forms of proprotein convertase subtilisin/kexin type 9 show that mutations in the LDL receptor, the ligand for the receptor, or an intracellular chaperone for the receptor are the most important determinants in regulating LDL catabolism. This review also demonstrates the variance of results within similar, or even the same, phenotypic conditions. This underscores the sensitivity of metabolic studies to methodological aspects and thus the importance of the inclusion of adequate controls in studies. PMID- 16720895 TI - Modulation of nuclear factor-kappaB by human T cell leukemia virus type 1 Tax protein: implications for oncogenesis and inflammation. AB - Activation of the nuclear factor kappa B (NF-kappaB) transcription factor family by different stimuli, such as inflammatory cytokines, stress inducers, or pathogens, results in innate and adaptive immunity. While the main function of NF kappaB is to promote the host's immune response, the NF-kappaB pathway is frequently dysregulated by invading viral pathogens. Human T cell leukemia virus type 1 (HTLV-1) is the causative agent of a fatal malignancy known as adult T cell leukemia (ATL) and an inflammatory disease named tropical spastic paraparesis/HTLV-1 associated myelopathy (TSP/HAM). HTLV-1 encodes an oncoprotein, Tax, which plays a significant role in the initiation of cellular transformation and the elicitation of the host's inflammatory responses. Here, we review current thinking on how Tax may affect both diseases through activation of NF-kappaB signaling. PMID- 16720897 TI - New players in TLR-mediated innate immunity: PI3K and small Rho GTPases. AB - Toll-like receptors (TLRs) play a crucial role in the innate immune system as a first line of defense against pathogens. TLR activation in phagocytes produces pro-inflammatory cytokines and chemokines that contribute directly to elimination of infectious agents and activation of adaptive immune responses. However, a sustained inflammatory response can result in tissue damage and generalized sepsis. This review summarizes the complex and sometimes conflicting links of TLR signaling with two important regulators of immune cells functions: phosphoinositide 3-kinases (PI3Ks) and small GTPases of the Rho family. A unified model of hierarchical organization of these signaling participants is still premature, given that the tools for delineating how control of TLRmediated pathways is achieved are just emerging. Critical progress in our understanding of spatial-temporal propagation of TLR signaling will certainly be provided in the near future by pharmacological targeting of PI3Ks using recently characterized, second-generation PI3K inhibitors in combination with gene-targeting strategies for PI3K subunits and Rho GTPases targeted to the murine myeloid compartment. PMID- 16720896 TI - Beyond mere markers: functions for CD34 family of sialomucins in hematopoiesis. AB - CD34, podocalyxin, and endoglycan are members of a family of single-pass transmembrane proteins that show distinct expression on early hematopoietic precursors and vascular-associated tissue. In spite of the fact that the expression of CD34 on these early progenitors has been known for over 20 yr and used clinically in hematopoietic stem cell transplantation for more than 15 yr, little is known about its exact role or function. More recently, CD34 expression has been shown to distinguish activated early progenitors from quiescent cells. With the subsequent identification of podocalyxin and endoglycan as related family members also expressed on early progenitor cells, attention is slowly shifting toward understanding how these molecules might contribute to progenitor function and behavior. In this review we examine the existing evidence and propose testable models to reveal the importance of these molecules for stem and progenitor cell function. PMID- 16720899 TI - Redirection of T cells by delivering a transgenic mouse-derived MDM2 tumor antigen-specific TCR and its humanized derivative is governed by the CD8 coreceptor and affects natural human TCR expression. AB - Retroviral transfer of T cell antigen receptor (TCR) genes selected by circumventing tolerance to broad tumor- and leukemia-associated antigens in human leukocyte antigen (HLA)-A*0201 (A2.1) transgenic (Tg) mice allows the therapeutic reprogramming of human T lymphocytes. Using a human CD8 x A2.1/Kb mouse derived TCR specific for natural peptide-A2.1 (pA2.1) complexes comprising residues 81-88 of the human homolog of the murine double-minute 2 oncoprotein, MDM2(81-88), we found that the heterodimeric CD8 alpha beta coreceptor, but not normally expressed homodimeric CD8 alpha alpha, is required for tetramer binding and functional redirection of TCR- transduced human T cells. CD8+T cells that received a humanized derivative of the MDM2 TCR bound pA2.1 tetramers only in the presence of an anti-human-CD8 anti-body and required more peptide than wild-type (WT) MDM2 TCR+T cells to mount equivalent cytotoxicity. They were, however, sufficiently effective in recognizing malignant targets including fresh leukemia cells. Most efficient expression of transduced TCR in human T lymphocytes was governed by mouse as compared to human constant (C) alphabeta domains, as demonstrated with partially humanized and murinized TCR of primary mouse and human origin, respectively. We further observed a reciprocal relationship between the level of Tg WT mouse relative to natural human TCR expression, resulting in T cells with decreased normal human cell surface TCR. In contrast, natural human TCR display remained unaffected after delivery of the humanized MDM2 TCR. These results provide important insights into the molecular basis of TCR gene therapy of malignant disease. PMID- 16720898 TI - The role of invariant natural killer T cells in lupus and atherogenesis. AB - Systemic lupus erythematosus (SLE) is increasingly recognized as a risk factor for the development of premature atherosclerosis. The inflammatory process in both of these diseases is controlled by a variety of cell types of the innate and adaptive immune systems. Recent studies from several groups, including ours, have revealed a critical role of a unique subset of lymphocytes, termed invariant natural killer T (iNKT) cells, in the development of lupus-like autoimmunity and atherosclerosis in animal models. iNKT cells appear to play complex and divergent roles in the development of SLE and atherosclerosis. Our findings suggest that alterations in iNKT cell functions during the development of SLE may be related to the increased risk of SLE patients to develop atherosclerosis and coronary heart disease. We found that iNKT cell activation with the sponge-derived glycolipid alpha- galactosylceramide generally protects against the development of lupus-like autoimmunity in mice, whereas it exacerbates atherosclerosis. Therefore, while our studies have identified iNKT cells as potential therapeutic targets for SLE, further studies are necessary to design drugs that will avoid the underlying harmful effects of iNKT cell activation on the development of atherosclerosis. PMID- 16720900 TI - Crosslinked penicillin acylase aggregates for synthesis of beta-lactam antibiotics in organic medium. AB - Crosslinked enzyme aggregates (CLEAs) of a partially purified penicillin acylase from a recombinant Escherichia coli strain have been produced as a novel type of biocatalysts well endowed to perform in organic media. Different protein precipitants were studied and glutaraldehyde was used as the crosslinking agent. Precipitation curves were obtained for all precipitants to determine the concentrations at which all the protein precipitated out of the solution. The effect of the glutaraldehyde-to-protein ratio was studied with respect to process recovery and the specific activity and stability of the biocatalyst. Recovery of penicillin acylase activity was moderately high, about 50%; major losses of enzyme activity were produced at the precipitation step. Specific activities of all CLEAs were very high, which is one of the advantages of using nonsupported biocatalysts. Ammonium sulfate and tert-butyl alcohol were the best precipitants at a glutaraldehyde-protein mass ratio of 2 and were selected to perform the kinetically controlled synthesis of ampicillin in 60% (v/v) ethylene glycol medium. At comparable conversion yields, volumetric and specific antibiotic productivity were much higher for CLEAs than for carrier-bound penicillin acylases. PMID- 16720901 TI - Production of ceramide with Saccharomyces cerevisiae. AB - The possibility of producing the biologically active material of the skin, ceramide, was studied using yeasts. The yeast strain that produced the most ceramide, Saccharomyces cerevisiae (KCCM 50,515), was selected, and the optimal conditions for ceramide production were determined using shakeflask culture and batch fermentation. By measuring the production rate of ceramide at various pH values and temperatures, the optimal conditions for ceramide production were found to be pH 6.0 and 30 degrees C. When heat shock was applied to the cells for 1 h by increasing the culture temperature from 30 to 40 degrees C after cell growth, the amount of ceramide produced was increased 5.9-fold. A cell growth and ceramide production model was developed with Monod kinetics and the Leudecking Piret model. It showed that ceramide production was increased when the cells were in the stationary phase. PMID- 16720902 TI - Structure-function relations of carbohydrates by neoglycolipid arrays. AB - The work presented herein is a new noncovalent glycoarray assembly method for microplates created by simply mixing together a carbohydrate and a tetradecylamine. alpha-D-Mannopyranoside, alpha-D-glucopyranoside, and alpha-D galactopyranoside were utilized in model studies and product formations were detected by lectin binding. The method can be extended to study the steric hindrance effect of carbohydrate-protein interactions, namely the structure function relations of carbohydrates. PMID- 16720903 TI - Recycling of distillery effluents in alcoholic fermentation: role in inhibition of 10 organic molecules. AB - In beet distilleries, condensates arising from stillage concentration could be recycled as dilution water for the fermentation step, thus preserving groundwater resources and ensuring a quality-controlled water supply. However, the recycling of condensates has been found to cause a significant reduction in fermentation activity. This study aimed to verify that condensates are toxic to alcoholic fermentation. Ten compounds found in condensates (formic, acetic, propionic, butyric, valeric, and hexanoic acids; 2,3-butanediol, furfuryl alcohol, furfural, and 2-phenyl-ethyl-alcohol) were tested. With the exception of 2,3-butanediol, they all proved to be inhibitors. At the same molar concentration, the longer the carbonaceous chain, the stronger the inhibition by fatty acids. An experimental design was used to study the inhibitory characteristics of the 10 compounds at the concentrations found in condensates. Synergistic effects were also confirmed. In real effluents, acetic acid was so highly concentrated that it became the strongest inhibitor. It is therefore necessary to eliminate it before recycling, as well as less concentrated compounds that may accumulate, as illustrated by the simulation. PMID- 16720904 TI - Phytase production by thermophilic mold Sporotrichum thermophile in solid-state fermentation and its application in dephytinization of sesame oil cake. AB - The phytase production by Sporotrichum thermophile TLR50 was recorded on all the commonly used animal feed ingredients tested to varying degrees in solid-state fermentation. Enzyme production increased to 180 U/g of dry moldy residue (DMR) in sesame oil cake at 120 h and 45 degrees C at the initial substrate-to-moisture ratio of 1:2.5 and aw of 0.95. Supplementation of sesame oil cake with glucose and ammonium sulfate further enhanced phytase titer (282 U/g of DMR). An overall 76% enhancement in phytase production was achieved owing to optimization. The mold secreted acid phosphatase, amylase, xylanase, and lipase along with phytase. By the action of phytase, inorganic phosphate was liberated efficiently, leading to dephytinization of sesame oil cake. PMID- 16720905 TI - Lactic acid production from cellulosic material by synergetic hydrolysis and fermentation. AB - The hydrolysis process on corncob residue was catalyzed synergetically by the cellulase from Trichoderma reesei and the immobilized cellobiase. The feedback inhibition to cellulase reaction caused by the accumulation of cellobiose was eliminated efficiently. The hydrolysis yield of corncob residue was 82.5%, and the percentage of glucose in the reducing sugar reached 88.2%. The glucose in the cellulosic hydrolysate could be converted into lactic acid effectively by the immobilized cells of Lactobacillus delbrueckii. When the enzymatic hydrolysis of cellulose and the fermentation of lactic acid were coupled together, no glucose was accumulated in the reaction system, and the feedback inhibition caused by glucose was also eliminated. Under the batch process of synergetic hydrolysis and lactic acid fermentation with 100 g/L of cellulosic substrate, the conversion efficiency of lactic acid from cellulose and the productivity of lactic acid reached 92.4% and 0.938 g/ (L.h), respectively. By using a fed-batch technique, the total concentration of cellulosic substrate and lactic acid in the synergetic process increased to 200 and 107.5 g/L, respectively, whereas the dosage of cellulase reduced from 20 to 15 IU/g of substrate in the batch process. The results of the bioconversion of renewable cellulosic resources were significant. PMID- 16720906 TI - Effect of aeration rate on production of xylitol from corncob hemicellulose hydrolysate. AB - The effects of different aeration conditions on xylitol production from corncob hemicellulose hydrolysate by Candida sp. ZU04 were investigated. Batch fermentations were carried out in a 3.7-L fermentor at 30 degrees C, pH 5.5, and agitation of 300 rpm. It was found that the two-phase aeration process was more effective than the one-phase aeration process in xylitol production. In the first 24 h of the aerobic phase, a high aeration rate was applied, glucose was soon consumed, and biomass increased quickly. In the second fermentation phase, aeration rate was reduced and an improved xylitol yield was obtained. The maximum xylitol yield (0.76 g/g) was obtained with an aeration rate of 1.5 vvm (KLa of 37 h-1) for the first 24 h and 0.3 vvm (KLa of 6 h-1) from 24 to 96 h. PMID- 16720907 TI - Efficient production of poly-gamma-glutamic acid by Bacillus subtilis ZJU-7. AB - A strain with high poly-gamma-glutamic acid (gamma-PGA) production was isolated from fermented bean curd, a traditional Chinese food. The strain was named Bacillus subtilis ZJU-7 according to 16s rDNA sequencing and its taxonomic characters. The culture conditions for gamma-PGA production were evaluated. The most suitable carbon and nitrogen sources were sucrose and tryptone, respectively. Exogenous L-glutamic acid was necessary for gamma-PGA production, and the production of gamma-PGA increased on the addition of L-glutamic acid to the medium. In the medium containing 60 g/L of sucrose, 60 g/L of tryptone, 80 g/L of L-glutamic acid, and 10 g/L of NaCl, the yield of gamma-PGA reached 54.4 g/L after cultivation at 37 degrees C for 24 h, which was the highest gamma-PGA production compared with values reported in the literature. The average molecular mass of gamma-PGA produced was about 1.24 x 106 Daltons. B. subtilis ZJU-7 is genetically stable and can synthesize levan instead of gamma-PGA without the addition of L-glutamic acid to the medium. PMID- 16720908 TI - Gastric mucosal proliferative and total tyrosine kinases activities increase in Helicobacter pylori-induced chronic gastritis. AB - BACKGROUND: The intestinal type of gastric cancer is thought to originate from cancer precursor lesions, progressing from H. pylori-induced chronic gastritis, atrophic gastritis, to intestinal metaplasia (IM) and dysplasia. Tyrosine kinases (tyr-k) represent the family of proteins that are widely expressed during cell metabolism and are considered as secondary markers for cellular proliferation and malignant transformation. AIM OF STUDY: The aim of the study was to evaluate the correlation between gastric mucosal histopathologic changes, total tyrosine kinases, and proliferative activities in patients with H. pylori infection. METHODS: Biopsy specimens from the gastric mucosa of 94 patients were assessed for H. pylori infection, histopathology (according to the Sydney classification), proliferative activity [Ki-67 immunohistochemistry with labeling index (LI) estimation], and total tyr-k activities (ELISA assay kit). RESULTS: Total tyr-k activities and Ki-67 LI were significantly higher in H. pylori (+) than H. pylori (-) group (728.1 +/- 175.3 vs 360.1 +/- 44.4 pmol P/mg/min. p <0,01 and 20.0 +/- 5.8 vs 10.9 +/- 1.3 %, respectively). A significant correlation has been observed between the Ki-67 LI and total tyr-k activities in patients with and without H. pylori infection. In cases of gastritis accompanied with atrophic changes or intestinal metaplasia in H. pylori (+) patients, Ki-67 LI and total tyr-k activities were particularly high compared to chronic gastritis without atrophy or intestinal metaplasia. CONCLUSION: Those results suggest that tyrosine kinases may play an important role in the development of gastric mucosal hyperproliferation in H. pylori-induced gastritis and possibly in early phase of gastric carcinogenesis. PMID- 16720910 TI - Long-term results after surgery for gastric cancer with or without jejunal reservoir: results of surgery for gastric cancer in Kanta-Hame central hospital in two consecutive periods without or with jejunal pouch reconstruction in 1985 1998. AB - BACKGROUND: Gastric cancer still has a disease-specific 5-yr survival less than 30% and an overall survival of about 15%. The quality of life of patients who undergo gastrectomy is poor owing both to the severity of the disease itself and to the mutilation of the upper gastrointestinal channel after the reconstruction. Therefore, the combination of a jejunal pouch with gastrectomy has been claimed to improve the life quality and nutritional status of these patients. AIM: To assess the clinical results after surgery for gastric cancer in two consecutive periods with or without jejunal-pouch reconstruction. METHODS: 271 consecutive patients referred for surgery for gastric cancer in 1985-1991 (116 patients) and in 1992-1998 (155 patients) in Kanta-Hame central hospital were retrospectively analyzed regarding their disease, mode of surgery, and the immediate and long term results. In the former observation period gastrectomy was performed with Roux-en-Y esophagojejunostomy without a reservoir, and in the latter period this procedure was combined with a jejunal reservoir. The data were collected from patient journals and from the death certificate obtained from the National Centre of Statistics in Finland. RESULTS: During the study period the incidence of cancer in the cardia increased among the surgical patients from 13.1 to 26.7% (p <0.05). Despite this proximal migration, the cancer-specific 5-yr survival remained practically unchanged during the two study periods, 29.4% and 32.2% (NS). During the period of jejunal-pouch reconstruction there were non significant increases of the incidences of local recurrence (from 18.9% to 26.5%), of immediate postoperative anastomotic fistulae (from 0.9% to 4.5%) as well as of the immediate mortality (from 2.6% to 3.7%) (NS for each). CONCLUSIONS: Despite proximal migration of gastric cancer and the application of a jejunal reservoir, the long-term as well as the immediate results after curative surgery (i.e., D2-gastrectomy) for gastric cancer have remained relatively unchanged. The jejunal-pouch reconstruction with the present technique after gastrectomy can therefore be safely applied. PMID- 16720911 TI - Colonic carcinoma resembling submucosal tumor: report of a case and review of the literature. AB - Submucosal tumor-like colorectal carcinoma, most of whose surface is covered with normal mucosa, is very rare. We report a case of colonic carcinoma resembling submucosal tumor. A 54-yr-old man visited our institution for an evaluation of a positive fecal occult blood test. Colonoscopic examination revealed a small, mainly red polypoid lesion with a central deep ulceration and many white spots in the sigmoid colon. Indigocarmine staining demonstrated that the white spots were faint shallow depressions. Magnifying colonoscopic examination showed that the lesion surface, except for the ulceration and the depressions, was covered with normal mucosa. Although the tumor was small, we strongly suspected its malignancy due to a deep ulceration. As we could not excise it endoscopically, we performed sigmoidectomy. The lesion was 12 mm in size. Histologic examination revealed that the lesion was a moderately differentiated adenocarcinoma that was mainly covered with normal mucosa, that carcinoma was exposed only at the ulceration and the depressions on the surface, and that it had expanded to the muscularis propria. Together with considerations from the literature, this type of colorectal carcinoma is supposed to be invasive and surgical resection should be considered, no matter how small it may be. PMID- 16720909 TI - Type I collagen and divalent cation shifts disrupt cell-cell adhesion, increase migration, and decrease PTHrP, IL-6, and IL-8 expression in pancreatic cancer cells. AB - BACKGROUND: We have shown in FG pancreatic cancer cells that alpha2beta1 integrin mediated type I collagen adhesion decreases parathyroid hormone-related protein (PTHrP), interleukin-6 (IL-6), and interleukin-8 (IL-8) expression, decreases the localization of E-cadherin and beta-catenin in cell-cell contacts, increases cell migration, and increases glycogen synthase kinase 3 (GSK3) and protein kinase B (PKB/Akt) phosphorylation states relative to alpha5beta1 integrin-mediated fibronectin (Fn) adhesion. AIM OF THE STUDY: To extend our observations in FG cells to other pancreatic cancer cell lines, and to determine whether E-cadherin mediated cell-cell adhesion and its downstream effectors were functionally involved in the ECM-mediated regulation of PTHrP, IL-6, and IL-8. METHODS: We used standard biochemical techniques to determine ECM-specific differences in E cadherin and beta-catenin localization, GSK3 and PKB/Akt phosphorylation, haptokinetic cell migration, and cytokine expression in pancreatic cancer cells. We also conducted functional studies using pharmacological inhibitors for GSK3 and PKB/Akt, as well as elevated Mg2+/Ca2+ ratios similar to pancreatic juice, and examined their effects on cytokine expression. RESULTS: Differences in E cadherin and beta-catenin localization along with GSK3 and PKB/Akt phosphorylation occur in multiple pancreatic cancer cell lines, resulting in differences in ECM-mediated haptokinesis and cytokine expression that are generally consistent with previous observations in FG cells. Our functional studies also suggest that E-cadherin-mediated cell-cell adhesion and downstream effectors are involved in PTHrP, IL-6, and IL-8 expression. CONCLUSIONS: These data indicate that alpha2beta1 integrin-mediated type I collagen adhesion disrupts cell-cell adhesion architecture, resulting in increased migration and decreased PTHrP, IL-6, and IL-8 expression in pancreatic cancer cells. PMID- 16720912 TI - Asymptomatic familial colon cancer with FDG-PET scanning for recurrent disease. AB - A 57-yr-old female was referred for screening colonoscopy because of a positive family history of colon cancer. A lobulated tumor mass was detected in the sigmoid colon. The resected specimen showed an invasive adenocarcinoma without lymph node involvement. Later colonoscopic evaluations and CT imaging failed to reveal definite evidence of recurrent disease but a late rising carcinoembryonic antigen level led to FDG-PET scanning and the detection of suspect lymph nodes in the retroperitoneum. Further histopathological and immunohistochemical evaluation of resected lymph nodes confirmed metastatic carcinoma from the primary colon carcinoma with extra-nodal spread. This case underscores the ongoing need for additional evidence-based studies on evolving imaging modalities used in the diagnosis and management of colonic cancer. PMID- 16720913 TI - Metastatic adenocarcinoma of the colon and follicular lymphoma within the same lymph node: a case report and review of the literature. AB - Concomitant adenocarcinoma and non-Hodgkin's lymphoma, both located in the intestinal tract, are unusual. We report a unique case of moderately differentiated of the cecum and a simultaneous follicular lymphoma, Grade 1, of the terminal ileum and regional lymph nodes in a 55-yr-old man. One lymph node was involved by both adenocarcinoma and follicular lymphoma. To our knowledge, this is the fifteenth reported case of concurrent adenocarcinoma and non Hodgkin's lymphoma of the intestine, but this is the first case with involvement of follicular lymphoma and adenocarcinoma within the same lymph node. PMID- 16720914 TI - Primary minute invasive de novo colonic adenocarcinoma appearing as submucosal tumor. AB - A 4 mm white-yellow submucosal tumor-like lesion was detected in the sigmoid colon of an asymptomatic 52-yr-old Japanese man. Because the lesion was unexpectedly suspicious for adenocarcinoma by pathological examination of the biopsy specimen, it was treated by endoscopic mucosal resection. The specimen obtained demonstrated well-differentiated adenocarcinoma without any adenomatous element, and was located principally in the submucosal layer with a maximum depth of 1600 mum from the muscularis mucosae. The cancer exposed to the luminal surface was pathologically concluded to be diminutive. Intriguingly, aggregation of lymphocytes was found beneath the mucosal layer, which might have compromised the integrity of the muscularis mucosae. Because of deep submucosal infiltration and the latent aggressive nature of de novo cancer, the patient underwent an additional partial sigmoidcolectomy, which demonstrated no residual cancer and no regional lymph node metastasis. The lesion in this patient exhibited a previously undescribed appearance of de novo colon cancer as submucosal tumor in an early phase of growth. PMID- 16720915 TI - Paclitaxel-carboplatin combination chemotherapy in advanced breast cancer: accumulating evidence for synergy, efficacy, and safety. AB - Patients with metastatic breast cancer receive multiple lines of cytotoxic chemotherapy, with taxane and anthracycline-based regimens being the most active. Anthracyclines carry the risk of significant cardiotoxicity at high cumulative doses and when combined with trastuzumab, an anti-HER2 antibody. Carboplatin has shown promising single-agent activity in advanced breast cancer, is not a P glycoprotein substrate, and is conveniently administered on an outpatient basis. Preclinical experiments demonstrated schedule-dependent synergistic cytotoxic effects of the paclitaxel first/carboplatin last (PC) combination. Pharmacokinetic parameters of paclitaxel and carboplatin were studied by Hellenic Cooperative Oncology Group (HECOG) and no significant interaction or correlation with clinical parameters were found. We assessed PC both as salvage as well as first-line treatment of advanced breast cancer patients in phase II studies which disclosed 40-60% response rates and median survival times of 12-20 mo with manageable toxicity. These results were confirmed by other groups and prompted us to the first randomized phase III trial comparing PC to the standard of epirubicin/paclitaxel (EP), a trial that showed equivalent efficacy and tolerable toxicity for PC. Registry retrospective analysis identified factors prognostic for improved outcome: good performance status, low tumor burden, lack of anthracycline exposure and of hormonal maintenance therapy. PC combinations with HER1 or HER2 modulators are being evaluated both by HECOG and by international groups. Paclitaxel coupled with carboplatin provides an alternative therapeutic option for anthracycline-exposed patients and warrants further clinical research in the direction of anthracycline-free management of metastatic breast cancer. PMID- 16720916 TI - Interstitial lung disease in patients with non-small-cell lung cancer treated with epidermal growth factor receptor inhibitors. AB - Interstitial lung disease (ILD) refers to a diverse range of pulmonary fibrotic disorders and may be hard to accurately diagnose, as distinguishing it from other pulmonary diseases can be difficult. Estimations of the incidence in populations are confounded by the complexity of the different forms of the disorder. In addition, ILD is a comorbid disease of lung cancer and is seen after most forms of chemotherapy and radiotherapy for advanced lung cancer. Incidences of >or=10% have been reported; however, whatever the true incidence, both chemotherapy and radiotherapy enhance the risk of developing ILD. ILD has also been reported with the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors, including erlotinib (Tarceva, OSI-774) and gefitinib (IRESSA). In a large number of gefitinib-treated patients (n > 185,000) an incidence of approx 1% has been observed (approx 2% in Japan; 0.3% in the rest of the world). Nevertheless, as with other treatments for advanced non-small-cell lung cancer, the clinical benefit outweighs the risk of ILD. In this article, we review the data on ILD with EGFR inhibitors and other common lung cancer treatments. PMID- 16720917 TI - Clinical and pathological predictors of the response to neoadjuvant anthracycline chemotherapy in locally advanced breast cancer. AB - PURPOSE: The aim of this study is to determine clinical and histopathological characteristics correlated to responsiveness to anthracycline-based neoadjuvant chemotherapy in breast cancer. PATIENTS AND METHODS: We studied primary tumor specimens with local advanced breast cancer from 40 patients. Patients received anthracycline-based chemotherapy. Neoadjuvant regimen consisted in 600 mg/m2 5 fluorouracil, 60 mg/m2 doxorubicin, and 600 mg/m2 cyclophosphamide (FAC). The World Health Organization criteria were used to classify the tumors. We performed immunohistochemical staining for ER, PgR, HER-2, PCNA (proliferation cell nuclear antigen), Ki-67, p53, and Bcl-2. Clinical and histopathological characteristics were associated with clinical response and histopathological changes induced by chemotherapy. RESULTS: The mean age was 47 +/- 14 yr. Twenty-three percent of patients were in stage IIB and 77% were in stages IIIA and IIIB. Seven percent of patients had progression of the disease. Stable disease was observed in 42% of patients and 45% had partial response. Only 7% of patients had a complete response. Factors associated with a better and major percentage of clinical response were the administration of doxorubicin-based chemotherapy, administration of more than three cycles, clinical N1, atypia, more than 10 mitosis per high-power field, moderate to severe SBR grade, and a major index of cellular proliferation. CONCLUSION: We found that tumors with large volumes, N2 node status, low cellular proliferation rate, positive immunoreactivity to p53, and low differentiation grade have a lower response to neoadjuvant chemotherapy with anthracycline. These patients could benefit from a different chemotherapy scheme to obtain a better control and resection. PMID- 16720918 TI - Effect of concomitant use of pentoxifylline and alpha-tocopherol with radiotherapy on the clinical outcome of patients with stage IIIB non-small cell lung cancer: a randomized prospective clinical trial. AB - We evaluated the effects of pentoxifylline (PTX) and alpha-tocopherol on the clinical outcome of 66 patients with stage IIIB non-small cell lung cancer in a randomized clinical trial. All patients received 46 Gy of external radiotherapy to the primary tumor and regional lymph nodes, with an additional 14-Gy dose to the primary tumor. Thirty-three of the 66 patients also received PTX (400 mg, three times daily) and alpha-tocopherol (300 mg, twice daily) during radiotherapy, followed by 400 mg of PTX and 300 mg of alpha-tocopherol daily for 3 mo after radiotherapy. The remaining 33 patients (control group) received radiotherapy only. After a mean follow-up time of 12 mo, 18 patients remained alive. During follow-up, there were local recurrences in 14 patients and distant metastases in 18 patients. In patients who received PXT and alpha-tocopherol, 1- and 2-yr overall survival rates were 55% and 30%, respectively, and median survival was 18 mo. In control patients, 1- and 2-yr overall survival rates were 40% and 14%, respectively, with a median survival of 10 mo. These differences were statistically significant (p = 0.0175). In patients who received PXT and alpha-tocopherol, progression-free survival rates for 1 and 2 yr were 48% and 23%, respectively; median survival was 12 mo. In the control group, the corresponding rates were 24% and 18%; median survival was 8 mo (p = 0.0223). We conclude that the use of PTX and alpha-tocopherol combined with radiotherapy offers a possible survival advantage in this patient population. PMID- 16720919 TI - Recombinant adenoviruses expressing TRAIL demonstrate antitumor effects on non small cell lung cancer (NSCLC). AB - INTRODUCTION: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis in a variety of malignant cells, but not in normal cells. This preferential toxicity to the abnormal cells renders TRAIL potentially a very powerful therapeutic weapon against cancer. However, a requirement for large quantities of TRAIL to suppress tumor growth in vivo is one of the major factors that has hindered it from being widely applied clinically. To overcome this, we constructed a replication-deficient adenovirus that carries a human full-length TRAIL gene (Ad-TRAIL) and tested its efficacy against a lung cancer model system in comparison to that of the recombinant soluble TRAIL protein. METHODS: To investigate the antitumor activity and therapeutic value of the Ad-TRAIL on the non-small cell lung cancer (NSCLC), four NSCLC cell lines, namely, YTMLC, GLC, A549, and H460 cells, were used. TRAIL protein expression was determined by Western blotting and flow cytometry. Cell viability was analyzed by proliferation assay, and DNA ladder and cell-cycle analysis were used to identify apoptosis. To further evaluate the effect of Ad-TRAIL in vivo, YTMLC cells were inoculated to the subcutis of nude mice. The Ad-TRAIL was subsequently administered into the established tumors. Tumor growth and the TRAIL toxicity were evaluated after treatment. RESULTS: YTMLC cells infected with Ad-TRAIL showed decreased cell viability and a higher percentage of apoptosis. Similar, Ad-TRAIL treatment also significantly suppressed tumor growth in vivo. CONCLUSIONS: TRAIL gene therapy provides a promising therapy for the treatment of NSCLC. PMID- 16720921 TI - The value of serum bcl-2 levels in advanced epithelial ovarian cancer. AB - The present study was conducted to investigate the value of serum bcl-2 levels in advanced epithelial ovarian cancer patients. Twenty-two patients with advanced ovarian carcinoma pathologically verified were investigated. Serum samples were obtained on the first admission before the chemotherapeutic treatment were given. Serum bcl-2 protein was determined by using ELISA. The baseline serum bcl-2 levels were significantly higher in patients with ovarian cancer than in the control group (p < 0.001). The sensitivity and specificity of serum bcl-2 were determined as 100% and 78%, respectively. None of the prognostic parameters analyzed such as age of patient, stage of disease, serum CA-125, albumin, hemoglobin, LDH, and response to chemotherapy was significantly correlated with bcl-2 serum concentrations. No prognostic value of serum bcl- 2 was determined. In conclusion, the results of the present study suggest that decreased apoptosis occurred due to the effect of serum bcl-2 elevation in advanced ovarian cancer patients. Also, serum bcl-2 level was a diagnostic but not a prognostic value in ovarian cancer. However, much researches still continues in this field, and exciting new knowledge will ultimately emerge. PMID- 16720920 TI - Comparison of osteopontin expression in endometrioid endometrial cancer and ovarian endometrioid cancer. AB - This study compared the DNA, RNA, and protein levels of osteopontin (OPN) in endometrioid endometrial cancer (EEC) and ovarian endometrioid cancer (OEC). In total, 63 cancer cases (EEC: 33, OEC: 30) were included. Of these, 47 (EEC: 26, OEC: 21) were examined by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and 48 (EEC: 25, OEC: 23) were examined by quantitative PCR. OPN expression was detected in 15 (50.0%) of 30 EECs and in 14 (50.0%) of 28 OECs. There was no significant difference in the percentage of positive cytoplasmic OPN staining between the EECs and OECs (12.8 vs 10.4; p = 0.6811). The correlation between relative mRNA and protein expression levels was significant in both the EECs and OECs; however, the correlation between relative DNA and mRNA levels was not significant. There was no significant difference in OPN expression between the EECs and OECs. PMID- 16720922 TI - Pelvic exenteration for recurrent or persistent cervical cancer: experience of five years at the National Cancer Institute in Mexico. AB - Cervical cancer constitutes a major health problem in Mexico and other developing countries. The purpose of our study was to assess the experience of a comprehensive national oncological reference center on pelvic exenteration for post-radiotherapy recurrent or persistent cervical cancer, describing the prognostic value of time to recurrence, procedure complications, and survival. Medical records from 42 patients with post-radiotherapy recurrent or persistent cervical cancer who underwent a pelvic exenteration with curative purposes from 1984 to 1989 were retrospectively reviewed. Histological diagnoses were squamous cell carcinoma (32 patients), adenosquamous carcinoma (9 patients), and adenocarcinoma (1 patient). Average follow up was of 56.3 mo after the procedure and global survival at 5 yr was 65.8%. Survival for patients with early recurrence was 56.9% vs 78% for patients with late recurrence (p = 0.05). Complications were observed in 65.3% of the cases with a surgical mortality of 4.8%. Pelvic exenteration is a surgical procedure with high morbidity in spite of the recent medical advances. Pelvic exenteration should not be indicated with palliative purposes owing to the high rate of complications. Patients with tumor persistence or early recurrence have a worse prognosis. In well-selected cases, exenteration may provide a survival benefit. PMID- 16720923 TI - Primary extranodal lymphomas of gastrointestinal localizations: a single institution 5-yr experience. AB - This study is aimed at comparison of patients with extranodal lymphomas based on pathohistological findings differences (MALT vs non-MALT) as well as regarding gastric and non-gastric localization, and determining the significance of clinical-laboratory parameters with respect to therapeutic response and length of survival. A total of 56 patients with extranodal non-Hodgkin's lymphomas of the gastrointestinal tract were evaluated over a 5-yr period. Regarding the localization of the disease, the stomach was most frequently affected, 39 patients (70%); followed by small and large intestines, 17 patients. As for the pathohistological findings, MALT lymphoma accounted for 70%, DLBCL 25%, while other subtypes accounted for 5%. Patients' distribution was analyzed according to CS based on both Ann Arbor and Lugano systems; however, the difference obtained between the groups was not statistically significant in both staging types of patients. Statistically significant difference in patients' distribution was not found with respect to IPI index, bone marrow infiltration, anemia, hypoalbuminemia, or histological subtype and localization. Difference in survival between patients according to the pathohistological type was not statistically significant also according to the type of the affected gastrointestinal tract organ. Statistical significance of difference according to survival probability was obtained based on age (survival is longer in patients over 55 yr of age); according to CS and according to Ann Arbor and Lugano classifications (the patients with lower CS live significantly longer); according to IPI index (the survival is significantly longer in patients with lower probability: IPI-0,1, and 2), as well as patients free of bone marrow infiltration whose survival is also significantly longer. PMID- 16720924 TI - Implications of CEA and p53 overexpression in the poor prognosis of colorectal cancer. AB - Colorectal cancer (CRC) is one of the most frequent and aggressive types of cancer. Several clinicopathologic features have been studied to identify the prognostic factors that can provide information concerning the favorable or the poor outcome of colorectal cancer. In the present study, the relationship between serum CEA, p53 expression, and DNA index to the different clinicopathological characteristics of colorectal cancer patients was sought. Fifty patients with CRC were included in this study. p53 protein was detected immunohistochemically using specific monoclonal antibodies. Samples were investigated for DNA index using flow cytometry. In addition, the serum CEA was determined using ELISA. The results showed that 27/50 (54%) were positive for p53. Concerning CEA reactivity, it was found that 35/50 (70%) were reactive for CEA. These results indicate that CEA is more sensitive than p53 to detect colorectal cancer. There was a statistically significant difference between the recurrent and nonrecurrent groups in the CRC Duke's stages, survival time, serum CEA (p = 0.001, 0.016, < 0.001, respectively). Kaplan-Meier method and log-rank test showed that the mean survival time for cases positive for both p53 and CEA is significantly different from cases positive for CEA only, positive for p53 only, and negative for both p53 and CEA (p = 0.0002). Survival time was statistically significant with respect to sex, p53, CEA, and Duke's stages (p = 0.006, 0.024, 0.001, 0.017, respectively). Cox regression model showed that the prognosis of colorectal cancer is influenced by sex, p53, CEA reactivity, and CRC Duke's stages (p = 0.014, 0.006, 0.019, 0.014, respectively). In conclusion, the use of more than one tumor marker may successfully aid in the prediction of colorectal cancer prognosis. PMID- 16720925 TI - Modulation of human renal cell carcinoma 786-0 MMP-2 and MMP-9 activity by inhibitors and inducers in vitro. AB - A hallmark of renal cell carcinoma (RCC) invasion is its ability to degrade ECM by local production of gelatinase enzymes. Although many studies on RCC have demonstrated the importance of MMPs, very little information is currently known regarding the effect of inducers and inhibitors. We therefore investigated the effect of inducers and inhibitors on RCC 786-0 in vitro. Human RCC 786-0 (ATCC) was grown in RPMI medium supplemented with 10% FBS, penicillin, and streptomycin in 24-well tissue plates. At near confluence, the cells were washed with PBS; the serum-free medium was incubated with various inducers: phorbol ester (PMA), tumor necrosis factor alpha (TNF-alpha), interleukin 1-beta (IL-1beta) and lipopolysaccharides (LPS). Cells were also incubated with inhibitors: EGCG, doxycycline, and a nutrient mixture with and without PMA; retinoic acid, dexamethasone, H-7; actinomycin D, or cyclohexamide. After 24 h, the medium was removed and analyzed for MMP-2 and MMP-9 by gelatinase zymography. RCC 786-0 secreted two bands, a major band corresponding to MMP-2 and a faint band corresponding to MMP-9. PMA and TNF-alpha, with increased concentration, increased MMP-9 secretion, while IL-1beta and LPS did not significantly modify MMP-9 activity. MMP-2 secretion was not affected by any of the inducers. All the inhibitors tested without and with PMA showed a dose-dependent decrease in both MMP-2 and -9 expression. Further studies are in progress to confirm the role of MMP-9 on Matrigel invasion using PMA, cytokines, and LPS. PMID- 16720926 TI - Prophylactic tropisetron versus rescue tropisetron in fractionated radiotherapy to moderate or high emetogenic areas: a prospective randomized open label study in cancer patients. AB - AIM: A prospective randomized open label study was carried out to evaluate the efficacy and effectiveness of prophylactic tropisetron versus rescue tropisetron in fractionated radiotherapy. PATIENTS AND METHODS: The study sample consisted of 288 cancer patients randomly allocated (3:4 ratio) into two treatment groups: 120 patients received prophylactic antiemetic treatment with tropisetron and 168 patients received rescue tropisetron. To determine the efficacy of prophylactic antiemetic treatment, nausea and vomiting were evaluated 1 d before radiation therapy (RT), at 24 and 72 h, at the end of every week during RT, and finally 1 wk after RT. Diary cards were used to record the intensity of nausea and vomiting as well as the incidence of adverse effects. RESULTS: In the odds of nausea and vomiting, statistically significant differences were found between the two treatment groups over time. The incidence of nausea and vomiting were 1.89 (p = 0.009) and 2.19 (p = 0.001) times higher in the rescue tropisetron group than in the prophylactic tropisetron group. Factors that related significantly with increased nausea were primary cancer, rescue tropisetron, and radical RT. Moreover, factors for vomiting were primary cancer type, metastasis, palliative RT, and rescue tropisetron. CONCLUSIONS: Higher numbers of patients receiving prophylactic tropisetron completed RT with lower incidence of nausea and vomiting than those in the rescue tropisetron group. PMID- 16720927 TI - Efficacy and safety results with the combination therapy of arsenic trioxide, dexamethasone, and ascorbic acid in multiple myeloma patients: a phase 2 trial. AB - BACKGROUND: Single-agent arsenic trioxide has shown promising results in patients with relapsed or refractory multiple myeloma (MM). Because preclinical data suggested greater activity with dexamethasone and ascorbic acid, a phase 2 trial of the combination of arsenic trioxide, dexamethasone, and ascorbic acid in patients with relapsed or refractory MM was conducted. METHODS: Twenty patients in whom no more than two previous therapies had failed were enrolled. The mean age was 62 yr, and 55% of the patients had refractory disease. The regimen consisted of 14- or 15-wk cycles, with the first cycle considered induction, followed by one or two consolidation cycles with a reduced steroid schedule and then a maintenance cycle in responding patients. RESULTS: The overall response rate was 30%, with at least stable disease in 80% of patients. Median progression free survival was 316 d in all patients and 584 d in those with a response. The regimen was well tolerated, with most adverse events being mild or moderate. CONCLUSIONS: This study showed the clinical efficacy and tolerability of the combination of arsenic trioxide, dexamethasone, and ascorbic acid. Further study is warranted. PMID- 16720928 TI - Dendritic cells, pulsed with lysate of allogeneic tumor cells, are capable of stimulating MHC-restricted antigen-specific antitumor T cells. AB - A variety of approaches have been used to deliver tumor-associated antigens (TAA) in conjunction with dendritic cells (DC) as cellular adjuvants. DC derived from monocytic precursors have been pulsed with whole tumor antigen using a variety of strategies and have been demonstrated to induce CD4+ and CD8+ antitumor responses. In the present study, monocyte-derived DC have been pulsed with lysate from an allogeneic melanoma cell line, A-375, and used to repeatedly stimulate T cells. The resultant T cells were examined for cytotoxic activity against A-375 targets as well as the HLA A2-positive melanoma cell line DFW. Uptake of FITC labeled melanoma lysate by DC established that lysate of melanoma cells was efficiently endocytosed. Stimulation with lysate-pulsed DC resulted in strong proliferative responses by T cells, which could be inhibited by antibodies against both MHC class I and class II. T cells stimulated in vitro with lysate pulsed DC demonstrated potent cytotoxicity against the melanoma targets which were blocked by antibodies against MHC class I. Lysate-pulsed DC also elicited IFN-gamma secretion by T cells as measured in an ELISPOT assay. We have also examined the ability of lysate-pulsed DC to present melanoma-associated antigens to T cells. ELISPOT assays with synthetic peptides of melanoma-associated antigens, such as gp100, mage1, NY-ESO, and MART-1, revealed that lysate-pulsed DC could stimulate T cells in an antigen-specific manner. The results demonstrate that lysate from allogeneic tumor cells may be used as a source of antigens to stimulate tumor-specific T cells in melanoma. PMID- 16720929 TI - Outcome for young high-risk aggressive B-cell lymphoma patients treated with CHOEP-14 and rituximab (R-CHOEP-14). AB - PURPOSE: To study the effectiveness and tolerability of a dose-intensified treatment including rituximab for patients, not older than 65 yr, with high-risk aggressive B-cell lymphoma. PATIENTS: Thirty-eight patients with high-risk aggressive B-cell lymphoma, the majority classified as grade 2 or 3 using age adjusted International Prognostic Index, were treated with six courses of CHOEP + rituximab on a 2-wk schedule with G-CSF d 4-11. CNS prophylaxis was administered using intravenous Ara-C as a single dose at the end of treatment. RESULTS: All patients were considered responders after three courses. Thirty-one patients (82%) achieved a complete remission or a complete remission unverified. With a median follow up of 27 mo, overall and event-free survival are 79% and 60%, respectively. Treatment was given on an outpatient basis. There were no treatment related unexpected toxic events or mortalities. Large-cell lymphoma involvement of the bone marrow was a poor prognostic sign even with this intensified treatment and 4/6 patients relapsed. CNS relapse occurred in three patients, two of whom had large cell bone marrow involvement. CONCLUSION: Although only a short follow up, the R-CHOEP-14 regimen is promising and could be an improvement compared to conventional treatment, with acceptable toxicity. The value of intravenous Ara-C at the end of treatment can be questioned, as it did not prevent CNS relapse or affect treatment outcome. PMID- 16720930 TI - Surgery and chemotherapy versus chemotherapy as treatment of high-grade MALT gastric lymphoma. AB - BACKGROUND AND OBJECTIVES: Treatment of high-grade MALT (mucosa-associated lymphoid tissue) gastric lymphoma remains uncertain. To assess efficacy and toxicity of the most common therapies--surgery followed by chemotherapy or chemotherapy alone--we began a controlled clinical trial in patients in early stage (I and II 1). METHODS: One hundred and two patients were randomized to be treated with surgery followed by six cycles of CEOP-Bleo (cyclophosphamide, epirubicin, vincristine, prednisone, and bleomycin at standard doses) (52 cases) or with chemotherapy alone (49 cases). RESULTS: Complete response rates were 94% [95% confidence interval (CI): 88-99%] and 96% (93-100%), respectively. Actuarial curves at 5 yr showed that event-free survival were 70% (95% CI: 59-74%) in patients treated with surgery and chemotherapy, that were not statistically significant to 67% (95% CI: 51-69%) in the patients who received chemotherapy (p = 0.5). Also, overall survival that was not statistically significant: 78% (95% CI: 70-88%) in the combined treatment and 76% (95% CI: 70-87%) in chemotherapy (p = 0.8). Acute and late toxicity were mild and well controlled. No acute leukemia or second neoplasm has been observed. CONCLUSIONS: The use of surgery and chemotherapy did not improve outcome in patients with early-stage high-grade gastric MALT lymphoma. It is apparent that chemotherapy alone is sufficient treatment in this select group of patients. PMID- 16720931 TI - Uterine choriocarcinoma in a postmenopausal woman. AB - Uterine choriocarcinoma developing in patients beyond reproductive age is a rare occurrence. We report a case of choriocarcinoma of uterine corpus in a 54-yr-old woman after 7 yr of menopause and 25 yr after last child birth. She presented with pain in the abdomen, and on radiological investigation a left uterine adnexal mass of 3.4 x 2.8 cm size was detected. Her serum CA125 level was 40 mIU/mL (normal up to 35 mIU/mL). Hysterectomy revealed an intramural growth in left uterine cornu measuring 3.5 x 3.0 x 2.5 cm. Histological features of the tumor were consistent with choriocarcinoma, and immunohistochemistry detected strong reactivity for beta-hCG in the tumor cells. Serum beta-hCG level 4 wk after surgery was 1345 mIU/mL. The patient was put on combination chemotherapy (EMACO). She achieved serological remission but showed a rise in serum beta-hCG level 4 wk after completion of chemotherapy. We conclude that a high level of suspicion may help in preoperative diagnosis of uterine choriocarcinoma in the postmenopausal age group. However, the response to chemotherapy in these cases may not be as encouraging as in choriocarcinoma of reproductive age. PMID- 16720932 TI - Pharmacological intervention for cognitive deficits and aggression in frontal lobe injury. AB - We present a case that promotes early intervention and pharmacological treatment for the neuropsychiatric sequelae (frontal lobe syndrome, including cognitive impairment and aggressive behavior) associated with traumatic brain injury (TBI) and delirium. The patient, who sustained significant systemic complications related to his trauma, was previously diagnosed with alcohol and drug dependence and antisocial personality disorder. These antecedent conditions and prolonged systemic complications likely played a complicating role in his course of recovery. PMID- 16720933 TI - Oculomotor rehabilitation for reading in acquired brain injury. AB - The purpose of this study was to assess reading-related oculomotor rehabilitation in individuals with acquired brain injury. Adults with either stroke (n=5) or traumatic brain injury (n=9) participated. Training paradigms included single line and multiple-line simulated reading, as well as basic versional tracking (fixation, saccade, and pursuit), twice per week over an 8 week period. Training modes included normal internal oculomotor visual feedback either in isolation (4 weeks) or concurrent with external oculomotor auditory feedback (4 weeks). Training effects were assessed objectively using infrared eye movement recording technology for simulated and actual reading, with the assessments occurring before, midway, and after training. In addition, the individuals were assessed subjectively using a reading rating-scale questionnaire. All reported considerably improved reading ability, and this was confirmed by several of the objective oculomotor measures. There was a trend for improvement to be better with the combined visual and auditory oculomotor feedback. Reading-related oculomotor rehabilitation produced significant gains in both the subjective and objective domains. It is believed that rapid saccadic oculomotor adaptation, as well as the training of rhythmicity and automaticity, were involved in modifying eye movement behavior to produce a more systematic approach and resultant improved reading profile. PMID- 16720934 TI - Drug induced arousal from the permanent vegetative state. AB - BACKGROUND: Zolpidem is an omega 1 specific indirect GABA agonist that is used for insomnia, but may have efficacy in brain damage. The long term efficacy of zolpidem in the permanent vegetative state is described in three patients. METHOD: Two motor vehicle accident patients and one near drowning patient, all of them in the permanent vegetative state for at least three years, were rated according to the Glasgow Coma and Rancho Los Amigos scale before and after zolpidem application. Long term response to daily application of this drug was monitored for 3-6 years. RESULTS: All patients were aroused transiently every morning after zolpidem. Glasgow Coma Scale scores ranged from 6-9/15 before to 10 15/15 after zolpidem. Rancho Los Amigos Cognitive scores ranged from I-II before to V-VII afterward. Drug efficacy did not decrease and there were no long term side effects after 3-6 years daily use. CONCLUSION: Zolpidem appears an effective drug to restore brain function to some patients in the permanent vegetative state. PMID- 16720935 TI - The effects of random whole-body-vibration on motor symptoms in Parkinson's disease. AB - It is well known that applying vibrations to men influences multiple physiological functions. The authors analysed post effects of whole-body vibration (WBV) on motor symptoms in Parkinson's disease (PD). Sixty-eight persons with PD were randomly subdivided into one experimental and one control group. Motor symptoms were assessed by the UPDRS (Unified Parkinson's Disease Rating Scale) motor score. A cross-over design was used to control treatment effects. The treatment consisted of 5 series of whole-body-vibration taking 60 seconds each. On average a highly significant (p<0.01) improvement of 16.8% in the UPDRS motor score was found in the treatment group. Only marginal changes (p>0.05) were found in the control group. The cross-over procedure showed comparable treatment effects (14.7% improvement after treatment). With respect to different symptom clusters only small changes were found in limb akinesia and cranial symptoms. By contrast, tremor and rigidity scores were improved by 25% and 24%, respectively. According to the structure of symptom changes it is unlikely that these effects are explainable on peripheral sensory level, exclusively. With respect to the findings of other studies one can speculate about changes in activation of the supplementary motor area and in neurotransmitter functions. PMID- 16720936 TI - Improved fatigue in individuals with multiple sclerosis after participating in a short-term self-care programme. AB - BACKGROUND: Fatigue is among the most common, yet least understood symptoms of Multiple Sclerosis (MS). The debilitating symptoms of MS can have adverse effects on the sufferer's self-esteem. We report the effect of a short-term self-managed graded exercise programme on fatigue and self-esteem of patients with MS. METHODS: Thirty-four (age range: 20-50, mean: 29.7 years; M:F 0.7:1.0) patients with MS who lived in Tehran and were not physically disabled entered the study. Self-esteem score was measured with the Persian translation of the Coopersmith Self-Esteem Inventory. Fatigue was evaluated with a visual analogue scale. The patients performed the self-managed techniques for 6 weeks. Self-esteem and fatigue were evaluated once more after the self-managed graded exercise programme. RESULTS: On paired sample test, the self-esteem score of the patients was significantly different before (53.9) and after (68.1) the self-managed graded exercise programme (P<0.0001). The score of fatigue was also significantly different before (4.59) and after (5.76) the intervention (P<0.0001). CONCLUSIONS: This study can provide us with important insights into our understanding of the potential for developing self-care training as an alternative in rehabilitation of the patients with MS. Self-managed graded exercise programme may be considered as an alternative to direct nursing services for patients with MS. PMID- 16720937 TI - Effects of dorsiflexor endurance exercises on foot drop secondary to multiple sclerosis: a pilot study. AB - PURPOSE: This is a pilot study to determine if endurance exercises for dorsiflexors will improve walking for people with foot drop secondary to MS, and if improvement in muscle endurance for persons with MS can be predicted based on the amount of central fatigue (CF) in the muscle. SUBJECTS: Five individuals with foot drop secondary to MS and five age-matched controls. METHODS: The intervention was 4 sets of 10 isometric contractions, at 60% of MVC, 3X/week, 8 weeks. Pretests-posttests included a dorsiflexor endurance test with superimposed electrical stimulation to measure CF, and a gait assessment including measurement of dorsiflexion at initial contact. RESULTS: Two of 5 subjects with MS had significant improvement in muscle endurance in both legs and in dorsiflexor control during walking. Correlation between CF and improved endurance was not significant (r=-0.07). One of the subjects with significant improvement in endurance had a significant decrease in central activation failure. CONCLUSIONS AND DISCUSSION: For individuals with foot drop secondary to MS, endurance exercises for the dorsiflexors can result in improved walking. CF may not be useful in predicting whether a muscle will improve in endurance with exercise. Improvement may result from either a peripheral training effect, a central learning effect, or both. PMID- 16720938 TI - The potential for neuropsychology to inform functional outcomes research with breast cancer survivors. AB - Cancer survivorship research has yielded important insights into the impact of breast cancer and associated treatments on such issues as vocational functioning, social role and community functioning, and quality of life. Efforts to understand factors that may impact these functional outcomes have specifically focused on medical, individual, and environmental variables. A relatively recent line of study suggests that neuropsychological functioning is an important individual variable to consider when assessing outcomes among breast cancer survivors, as there is evidence that at least a portion of women undergoing chemotherapy treatment for breast cancer will evidence an apparent decline in neuropsychological functioning. Two critically important issues are less well understood at this time, however. First, what is the underlying etiology of observed changes in cognitive functioning? Second, what is the functional significance of changes in neuropsychological functioning on work, social role and quality of life outcomes? To highlight potential directions for future research, this paper provides a review of existing literature documenting theories and research addressing these issues, and it discusses the potential impacts that changes in neuropsychological abilities may have on vocational, social role, and quality of life outcomes among breast cancer survivors. PMID- 16720939 TI - Neurogenic bladder dysfunction in patients with neoplastic spinal cord compression: adaptation of the bladder management strategy to the underlying disease. AB - OBJECTIVES: To study the outcome of different bladder management strategies in patients with neoplastic spinal cord compression. METHODS: 22 patients with neoplastic spinal cord compression underwent urodynamic examination. According to the urodynamic data and the underlying disease different bladder management strategies were recommended. In patients with curatively treated disease a full bladder rehabilitation program was arranged. In patients with metastatic malignant disease, voluntary voiding was continued if possible or a suprapubic catheter was placed. RESULTS: Eight patients were treated with curative intention. Of those, 2 patients were able to void during urodynamics continued normal voluntary voiding. Six patients were taught intermittent catheterisation, with three additionally received oral anticholinergic treatment because of UMN lesion. At follow-up, all patients had successfully finished bladder rehabilitation program and at follow up, all patients were continuing their previously recommended bladder rehabilitation program. Fourteen patients had malignant disease and were treated palliatively. In 2 patients with UMN lesion, voluntary control of micturition was maintained and both continued voluntary voiding. In 12 patients a suprapubic catheter was inserted. At follow-up, 9 out of 14 had died (mean 8 month after primary visit), the remaining 5 continued treatment with suprapubic catheters. CONCLUSION: Underlying disease and life expectancy should be considered for the selection of bladder management in patients with neoplastic spinal cord compression. In patients with curatively treated disease, a full bladder rehabilitation program is recommended while in patients with malignant disease and palliative care, a suprapubic catheter might be the treatment of choice. PMID- 16720940 TI - Tutorial on maximum inspiratory and expiratory mouth pressures in individuals with idiopathic Parkinson disease (IPD) and the preliminary results of an expiratory muscle strength training program. AB - Respiratory symptoms are recognized as sequelae of motor dysfunction in idiopathic Parkinson's disease (IPD) and these symptoms have the potential to cause problems with swallow, cough, voice and speech. Specifically, maneuvers that require rapid activation and coordination of upper airway and chest wall musculature become progressively impaired as motor dysfunction progresses during the natural course of the disease. This study reports on the maximum inspiratory and expiratory pressures produced by 28 participants (average age 64) diagnosed with moderate to severe IPD (average stage 2.5 with a range of 2.0-3.0). All measures were collected during the "medication on" state. Outcomes of a specific respiratory muscle strength training technique for improving maximum expiratory pressure are reported for three of the patients in this study. Techniques that focus on strengthening the respiratory muscles in patients with IPD (other than with low load breathing exercises), have not been previously reported. The results of this pilot study demonstrate that respiratory muscle weakness may be an important factor in the respiratory complications in IPD and that respiratory muscle strength training has the potential to improve expiratory muscle strength for this population. This improvement has the potential to positively impact high forced respiratory activities, such as forced breathing maneuvers, swallow, cough and speech functions that require greater magnitude and duration of expiration. PMID- 16720941 TI - To drive or not to drive (after TBI)? A review of the literature and its implications for rehabilitation and future research. AB - Development of reliable procedures to assess fitness to safe driving after traumatic brain injury (TBI) is a crucial step in rehabilitation. However, prior studies are highly inconsistent in the choice of measures recommended for predicting driving fitness from different pre-driving measures. In the present paper the relevant literature is reviewed with the aim of shedding light on the reasons for these inconsistencies. The discrepant results reflect investigative choices which differ in five aspects: (1) the type of predictors used as pre driving screening; (2) the type of measures considered as the criterion for the determination of fitness to drive after TBI; (3) the severity of the TBI in the sample of patients studied; (4) the extent of the neural structures damaged by TBI and the overlap of these areas with those involved in driving tasks; (5) the length of the follow-up considered. The strengths and weaknesses of the different methods and measures are discussed with their implications for future research and clinical rehabilitation. Encouraging findings come from recent studies that combined together medical, psychosocial, and personality measures, thereby improving the explanatory power of the predictors used. The use of post-injury driving fitness measures with great ecological and external validity seems equally promising in assessing actual driving in the real world. PMID- 16720942 TI - Peri-infarct reorganization in a patient with corona radiata infarct: a combined study of functional MRI and diffusion tensor image tractography. AB - PURPOSE: We investigated the motor recovery mechanism of a stroke patient using functional MRI (fMRI) and diffusion tensor image tractography (DTT). METHODS: A 60 year-old woman with right corona radiata infarct presented with complete paralysis of the left upper extremity. The motor function of the affected upper extremity recovered slowly for 6 months to a point of nearly normal function. The motor function of the patient was evaluated by fMRI and DTT at 6 months from the onset. FMRI was performed at 1.5 T with timed hand grasp-release movements. DTT was performed using 1.5 T with a synergy-L Sensitivity Encoding head coil. Three dimensional reconstructions of the fiber tracts were obtained with FA < 0.3, angle > 45 degrees as termination criteria. RESULTS: The contralateral primary sensorimotor cortex (SM1) was activated during both affected (left) and unaffected hand movements. DTT showed that the tracts originated from either SM1; the tracts then descended along the known pathway of the corticospinal tract, with the exception that the tract of the affected (right) hemisphere descended along the posterior portion of the infarcted corona radiata. CONCLUSIONS: It seems that the motor function of the affected upper extremity was reorganized to the posterior portion of the infarcted corona radiata. This finding may reflect functional reorganization of the motor pathway following corticospinal tract damage. This appears to be one motor recovery mechanism of subcortical infarct. PMID- 16720944 TI - Statin-mediated protective effects in the central nervous system: general mechanisms and putative role of stress proteins. AB - PURPOSE: The 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, or "statins", are used as cholesterol-lowering agents worldwide. This review, focused on recent experimental and clinical data, summarizes general mechanisms of statin actions underlying neuroprotective effects in the central nervous system (CNS) and presents evidence for putative mechanisms involving heat shock proteins and the survival-related PI-3K/Akt pathway that may be beneficial for the treatment of neurological disorders. METHODS: We carried out a medline search on statin actions, with respect to biochemical pathways and signal transduction on the one hand, and clinical studies in neurologic and retinal diseases on the other hand. Novel experimental data obtained in a rat model of axonal CNS injury were also included. RESULTS: Statins exert multiple effects on endothelial function, cell proliferation, inflammatory response, immunological reactions, platelet function, and lipid oxidation. These "pleiotropic actions" are independent of cholesterol lowering and appear to be beneficial in the context of brain injury. Several mechanisms of statin actions underlying neuroprotective effects may also involve heat shock proteins and the survival related PI-3K/Akt pathway. CONCLUSIONS: Available data suggest that statins may be of potential therapeutic use in a variety of diseases of the CNS including ischemic stroke, Alzheimer's disease, multiple sclerosis and some forms of retinal and eye diseases. Before general recommendations can be made and specific therapeutic approaches can be developed, more reliable clinical data and studies are required, and possible side effects must be carefully evaluated. PMID- 16720945 TI - Combination of TMS and fMRI reveals a specific pattern of reorganization in M1 in patients after complete spinal cord injury. AB - PURPOSE: After a spinal cord injury (SCI), which was complete, deafferentation of the body representation caudal to the lesion height results in drastic changes in the cortical representation. The underlaying processes are poorly understood. METHODS: We investigated cortical representation sites of upper limb muscles using functional magnetic resonance imaging (fMRI) and transcranial magnetic stimulation (TMS) in five patients suffering from thoracic complete SCI and one with an incomplete SCI in the height of L1. RESULTS: In comparison to healthy controls fMRI demonstrated a displacement of elbow movement representations in the precentral gyrus in patients with complete SCI into the direction of the deafferented cortical thoracic representation. Changes increased with time after the incidence of SCI. TMS revealed reduced excitability and prolonged silent periods for muscles more distant to the deafferented area. CONCLUSIONS: Whereas fMRI demonstrated changes in representation sites adjacent to the deafferented area, TMS excitability changes were also observed more distant to the deafferented area and silent periods were prolonged in comparison to healthy controls. TMS changes might depend on both: the distance to the deafferented area and the time of persistence of deafferentation. PMID- 16720943 TI - Spinal reflex control of micturition after spinal cord injury. AB - PURPOSE: This review summarizes possible spinal reflexes involved in micturition control as well as current treatments for lower urinary tract dysfunction after spinal cord injury (SCI) in an effort to further promote the development of new treatments for people with SCI. METHODS: A systematic literature search was conducted on the Medline database, and on the Web of Science. RESULTS: After SCI normal functions of the lower urinary tract are lost. The detrusor contracts frequently even at a small bladder volume, which results in urinary incontinence. In addition, the hyperactive bladder can not empty well because the urethral sphincter contracts simultaneously with the bladder (detrusor sphincter dyssynergia), which results in a large residual volume of urine. The malfunction of the lower urinary tract after SCI is caused by remaining and newly emerged bladder and urethral sphincter spinal reflexes. Current treatments have either limited success or troublesome side effects, and many new treatments are under development. CONCLUSIONS: In order to restore the functions of the lower urinary tract after SCI, efforts should be directed at re-establishing the coordination between the spinal reflex control of bladder and urethral sphincter using either pharmacological or neurophysiological approaches. PMID- 16720947 TI - Stimulation of the basolateral amygdala improves the acquisition of a motor skill. AB - PURPOSE: We have previously shown that the stimulation of limbic structures related to affective life such as the amygdale can improve and reinforce neural plastic processes related to hippocampus-dependent forms of explicit memory, as spatial memory and LTP. We now assessed whether this effect is restricted to the mentioned structure and memory type, or represents a more general form of modulatory influence. METHODS: Young, male Sprague Dawley rats were implanted stereotactically with one electrode in the basolateral amygdala (BLA) and trained to acquire a motor skill using their right anterior limb. A group of animals received 3 trains of 15 impulses at the BLA 15 minutes after each daily training session. A second group of implanted animals was handled in the same way, but not stimulated, while a third group was not implanted. After reaching the training criterion the left motor cortex was mapped by the observation of the movements induced by stimuli applied in discrete points of the cortex. RESULTS: Cortical representation of the anterior limb was increased in all trained animals, showing that the motor cortex is involved in the acquisition of the new skill. Animals receiving stimulation of the BLA showed similar cortical changes, but learned faster than non-stimulated controls. CONCLUSIONS: Reinforcement of neural plasticity by the activation of the amygdala is not restricted to hippocampus dependent explicit memory, but it might represent a universal mechanism to modulate plasticity. PMID- 16720946 TI - Cognitive evaluation of traumatically brain-injured rats using serial testing in the Morris water maze. AB - PURPOSE: As deficits in memory and cognition are commonly observed in survivors of traumatic brain injury (TBI), causing reduced quality of life for the patient, a major goal in experimental TBI research is to identify and evaluate cognitive dysfunction. The present study assessed the applicability of the serial Morris water maze (MWM) test to determine cognitive function following experimental TBI in the same group of rats which is particularly important for long-term studies and increasingly valuable for the evaluation of novel treatment strategies. METHODS: Male Sprague-Dawley rats (n = 27) were anesthetized and subjected to either sham injury (n = 9) or lateral fluid percussion (FP) brain injury of moderate severity (n = 18). At 4 weeks post-injury, animals were trained in a water maze over 3 days (acquisition/learning phase) to find a submerged platform. At 8 weeks post-injury the hidden platform was then moved to the opposite quadrant, and animals were trained to find the new position of the platform over 3 days. Forty-eight hours later, animals were tested for memory retention in a probe trial in which the platform was not present. RESULTS: Brain-injured animals had significant learning impairment (p < 0.0001), shifted-learning impairment (p < 0.001) and memory retention deficits (p < 0.01) in comparison to their sham injured counterparts over the 8 week testing period. Swim speed and distance were not significantly altered by brain injury at any time point. CONCLUSION: The validation of this testing paradigm using a clinically relevant experimental brain injury model is an important addition to behavioral outcome testing. PMID- 16720948 TI - Functional brain imaging, clinical and neurophysiological outcome of visual rehabilitation in a chronic stroke patient. AB - PURPOSE: To study rehabilitation outcome in a chronic visual field defect patient using positron emission tomography (PET) imaging, clinical, and neurophysiological measures. METHODS: A patient with chronic visual field defect was rehabilitated using a computer program over a three-month period. Evaluations of rehabilitation effects were done before and after training, and after a three month follow-up with perimetry, visual evoked potentials (VEP), subjective questionnaire, and PET activation studies of regional cerebral blood flow (rCBF). RESULTS: In perimetry, the visual field increased during the training, and the patient also experienced subjective improvement. In VEP, a previously absent P100 component was identified after the training. In PET, increases in rCBF were diffuse immediately after the training, and more occipital after the late follow up. A significant positive correlation between the increase in rCBF and improvement in clinical measures was found in the lingual gyrus in the contralesional hemisphere. CONCLUSIONS: All clinical measures showed improvement after rehabilitation, and this was maintained during the follow-up. In PET, a more generalized effect, found at the beginning, later concentrated in the occipital cortex. The results suggest that visual training may cause plastic changes in the brain, and that these changes can be detected both with clinical measures and with functional imaging. PMID- 16720949 TI - An analytical model for the investigation of axial wall forces generated by an arterial stenosis. AB - The causes of arteriosclerosis are intensively investigated since many decades. While circumferential wall stress has received a lot of attention, axial stress (also called "longitudinal" stress) has been largely neglected, and practically never incriminated. However, it has been suggested in 2003 that moderate and severe arterial stenoses may induce non negligible axial forces cyclically in the vessel segment just proximal to the constriction cone. In the present contribution, we describe a simple analytical model that allows to study the distribution of these forces along the vessel in dependence of the respective axial elasticities of vessel and surrounding tissues, and of the stenosis length. PMID- 16720950 TI - Information technology infusion model for health sector in a developing country: Nigeria as a case. AB - To date, information technology (IT) has not been widely adopted in the health sector in the developing countries. Information Technology may bring an improvement on health care delivery systems. It is one of the prime movers of globalization. Information technology infusion is the degree to which a different information technology tools are integrated into organizational activities. This study aimed to know the degree and the extent of incorporation of Information Technology in the Nigerian health sector and derive an IT infusion models for popular IT indicators that are in use in Nigeria (Personal computers, Mobile phones, and the Internet) and subsequently investigates their impacts on the health care delivery system in Nigerian teaching hospitals. In this study, data were collected through the use of questionnaires. Also, oral interviews were conducted and subsequently, the data gathered were analyzed. The results of the analysis revealed that out of the three IT indicators considered, mobile phones are spreading fastest. It also revealed that computers and mobile phones are in use in all the teaching hospitals. Finally in this research, IT infusion models were developed for health sector in Nigeria from the data gathered through the questionnaire and oral interview. PMID- 16720951 TI - Measuring dissimilarity in online health search activities. AB - There is growing evidence of the use of the Internet to obtain critical health information as well as increased diversity of user groups. While users of web based information often report their reliance on healthcare providers for validation of information, it has yet to be determined if this is universally true across all types of information or user groups. In some cases consumers may aggregate in places where they trust web information (relative to provider-based information), such as areas of unsettled diagnostic methods or ambiguous treatment protocols. To date little research has been done to identify and differentiate clusters of health consumers and their similarities related to type of information sought. Data from a study of consumer Web search activity in a post-intervention era serves as a natural experiment, and can identify whether clusters of "digitally underserved groups" persist in the US, following national efforts to eliminate barriers to health information access. This exploratory technology assessment study seeks to differentiate and delineate specific information behaviors, across targeted healthcare subgroups. Doing so allows the design of more effective strategies to promote the use of the Web as a health education and health promotion tool, under the envisioned shared decisionmaking, consumer-centric health information model, critical to the proposed US national health information infrastructure. PMID- 16720952 TI - New technology and old habits: The role of age as a technology chasm. AB - Familiarity with and use of computers is increasingly cited as a limiting factor related to the potential of the Internet as an effective healthcare resource for the elderly. This study sought to examine differences across age groups in their access to and use of computers and the Internet as a health information resource. We perform multivariate analysis with data from a stratified sample of survey reported health information-seeking behaviors on the Internet. The results suggest that access to online health information among Internet users varies significantly from the use of computers and Internet. Younger participants showed the highest rates in the use of computer and Internet, and exhibited the lowest rates in the access to online health information. In 2000, 56.7% and 55.9% of medium and older Internet users ever looked for health information on the Web, respectively, whereas 49.4% of young Internet users ever did. By 2002, these fractions had increased by 14.6%, 13.5% and 9.1%, respectively. The comparison of ORs between 2000 and 2002 indicated that the gap is somewhat increased, although the increase was not statistically significant at 0.05 level (the ORs of old to young at2000 = 1.30 [0.933, 1.817]; the ORs at2002 = 1.61 [1.157, 2.255]). Findings show that the gap between old and young seekers users of computers and the Internet for information appears to have widened in recent years, even as advocates of consumer-centric healthcare continue to promote the Internet as a key source of empowerment for the elderly. PMID- 16720953 TI - Pulse transit time as an indirect marker for variations in cardiovascular related reactivity. AB - Study of cardiac functions has shown to be important in understanding the prognosis and pathogenesis of vascular diseases. A physiologic parameter termed pulse transit time (PTT) has been studied intensively to be used as an indirect marker of such pathologic conditions. It is defined as the time it takes a pressure pulsation to travel between two arterial sites. For convenience of measurement, the interval between the peak of the R-wave on the electrocardiogram and the onset of the corresponding pulse in the finger pad measured by photoplethysmography is usually used. In this review, its definition, current applications in cardiovascular related studies and issues are presented in turn. Investigational studies suggest that PTT did not meet the level of accuracy required for medical practice as a marker for beat-to-beat blood pressure (BP). However, documented evidences indicate that PTT has high correlations when it is utilised as a surrogate semi-quantitative monitor of abrupt BP changes. Present literature suggests that description of disease-specific cardiovascular reactivity pattern is possible with techniques based on PTT. The attraction of PTT due to its non-invasive nature and revealing properties can only increase. Hence, the use of this simple technique in a clinical setting is still to realise its full potential. PMID- 16720954 TI - Computer-assisted, fluoroscopy-based ventral spondylodesis of thoracolumbar fractures. AB - OBJECTIVE: To design and evaluate a novel computer-assisted, fluoroscopy-based planning and navigation system for minimally invasive ventral spondylodesis of thoracolumbar fractures. MATERIALS AND METHODS: Instruments and an image intensifier are tracked with the SurgiGATE navigation system (Praxim-Medivision). Two fluoroscopic images, one acquired from anterior-posterior (AP) direction and the other from lateral-medial (LM) direction, are used for the complete procedure of planning and navigation. Both of them are calibrated with a custom-made software to recover their projection geometry and to co-register them to a common patient reference coordinate system, which is established by attaching an opto electronically trackable dynamic reference base (DRB) on the operated vertebra. A bi-planar landmark reconstruction method is used to acquire deep-seated anatomical landmarks such that an intraoperative planning of graft bed can be interactively done. Finally, surgical actions such as the placement of the stabilization devices and the formation of the graft bed using a custom-made chisel are visualized to the surgeon by superimposing virtual instrument representations onto the acquired images. The distance between the instrument tip and each wall of the planned graft bed are calculated on the fly and presented to the surgeon so that the surgeon could formalize the graft bed exactly according to his/her plan. RESULTS: Laboratory studies on phantom and on 27 plastic vertebras demonstrate the high precision of the proposed navigation system. Compared with CT-based measurement, a mean error of 1.0 mm with a standard deviation of 0.1 mm was found. CONCLUSIONS: The proposed computer assisted, fluoroscopy-based planning and navigation system promises to increase the accuracy and reliability of minimally invasive ventral spondylodesis of thoracolumbar fractures. PMID- 16720956 TI - Current and emerging pharmacological treatment options for dementia. AB - Treatments for the symptomatic relief of Alzheimer's disease are available but despite advances in our ability to treat persons with various forms of dementia, more effective treatments are needed. The cholinesterase inhibitors donepezil, rivastigmine, and galantamine have demonstrated efficacy in improving cognition and global status and to a lesser extent, behavioral abnormalities relative to placebo in patients with mild-to-moderate Alzheimer's disease. Rivastigmine has been shown to benefit patients with dementia with Lewy Bodies and with dementia associated with Parkinson's disease. Donepezil and galantamine have also been shown to be mildly effective in dementia due to cerebral ischemia. Memantine has a distinct mechanism of action and is effective in moderate-to-severe AD. The benefits from these drugs, however, are limited and their long-term effectiveness has not been well-demonstrated. Their clinical utility is controversial. Many novel approaches that promise to provide more effective treatments are currently being pursued. PMID- 16720957 TI - Neuroprotection and stroke rehabilitation: modulation and enhancement of recovery. AB - Recent advances in research are modifying our view of recovery after nervous system damage. New findings are changing previously held concepts and providing promising avenues for treatment of patients after stroke. This review discusses mechanisms of neuronal injury after brain ischemia and the attempts to study neuroprotection options based on such mechanisms. It also considers measures available at present to improve outcome after stroke and presents new areas of research, particularly stimulation techniques, neurogenesis and trophic factors to enhance recovery. In order to improve outcomes, medications that may be detrimental to recovery should be avoided, while symptomatic therapy of problems such as depression, pain syndromes and spasticity may contribute to better results. Continued surveillance and early treatment of complications associated with acute stroke, along with supportive care remain the mainstay of treatment for stroke patients in the recovery phase. Present research on limiting brain damage and improving recovery and plasticity enhance the prospects for better clinical treatments to improve recovery after stroke. PMID- 16720958 TI - Pharmacotherapy of posttraumatic cognitive impairments. AB - Pharmacotherapy may contribute to the rehabilitation of persons with posttraumatic cognitive impairments. This article reviews first the neurobiological consequences of traumatic brain injury with a particular emphasis on acute and long-term posttraumatic neurochemical disturbances. Studies of pharmacotherapies for posttraumatic cognitive impairments are reviewed next, and are organized according to medication class and the neurotransmitter system they affect most. Based on the evidence provided by that review, augmentation of posttraumatic cerebral catecholaminergic and cholinergic function are suggested as potentially useful neurochemical targets for pharmacologic intervention in this population. More specifically, it is suggested that persons with posttraumatic impairments in arousal, speed of processing, and possibly attention may benefit most from treatment with an agent that augments cerebral catecholaminergic function, and that persons whose predominant posttraumatic impairment is in the domain of memory may benefit most from treatment with cholinesterase inhibitors. Practical considerations regarding the use of pharmacotherapies for posttraumatic cognitive impairments are offered, and the need for additional research in this area is highlighted. PMID- 16720959 TI - Neuropharmacological treatment of mental dysfunction in Parkinson's disease. AB - Many patients with Parkinson's Disease (PD) experience significant cognitive and mood impairment -even early in the course of the disease. These mental impairments are only partially responsive to levodopa treatment and are often as disabling as the motor impairment, particularly in mid and late stages of the disease. Investigators have recently begun a search for new agents that can effectively treat mental dysfunction of PD. Although there have been only a handful of properly controlled clinical trials of interventions targeted at amelioration of mental dysfunction in PD, progress has been made. Based on the available evidence, targeting catecholaminergic and cholinergic function may be an effective strategy for amelioration of cognitve, mood and psychiatric disturbances in PD. PMID- 16720960 TI - Treatment of cognitive impairment in multiple sclerosis. AB - Cognitive impairment in multiple sclerosis is an increasingly recognized entity. This article reviews the cognitive impairment of multiple sclerosis, its prevalence, its relationship to different types of multiple sclerosis, and its contribution to long-term functional prognosis. The discussion also focuses on the key elements of cognitive dysfunction in multiple sclerosis which distinguish it from other forms of cognitive impairment. Therapeutic interventions potentially effective for the cognitive impairment of multiple sclerosis are reviewed including the effects of disease modifying therapies and the use of physical and cognitive interventions. PMID- 16720961 TI - Neuropharmacological aspects of cognitive neurorehabilitation in epilepsy. AB - The role of CNS neuromodulators in cognitive neurorehabilitation can be related to two main issues: 1) the negative impact on cognition of drug categories prescribed for different neurologic symptoms, such as spasticity, extrapyramidal symptoms, or epileptic seizures; 2) their possible role in neuroprotection and amelioration of the cognitive status of the patient, especially attention and memory. This paper reviews different pharmacological aspects of cognitive neurorehabilitation in epilepsy. PMID- 16720962 TI - Job matching and return to work: Occupational rehabilitation as the link. AB - Return to work after injury or illness is important for the worker and the employer. Medical providers manage and treat the worker with the illness or injury. Except in cases of focused specialists, the medical professional's role is to take care of a patient, rather than empower a worker. As much as there is promotion of the workers compensation health care system to be similar to sports medicine, there are significant dissimilarities. One major barrier is that the medical caregivers do not know the demands of jobs as they would know the details of sports. Thus, there is a gap in returning a worker to function as the medical professional cannot accurately match the worker to specific jobs. A new model of job function matching, based on research and skills of occupational rehabilitation professionals, is proposed to bridge the gap between the medical community, the employers and the workers. PMID- 16720963 TI - Consulting in industry: Moving beyond traditional interventions. AB - Despite increased efforts to address musculoskeletal disorders (MSDs) in the work place, these injuries continue significantly impact the bottom line financial performance of companies, not to mention the health and safety of their most important resource - their workers. As MSD costs continue to rise, rehabilitation professionals must approach effective injury management and prevention processes from an onsite perspective. A paradigm shift is in order. Successful practitioners need to transform themselves from a clinical specialist to an onsite consultant. This article introduces various elements that are critical to the successful transformation of an onsite consultant. Effective onsite injury management and prevention concepts are discussed. Case studies are also presented to further illustrate the value of the onsite consultant approach. PMID- 16720964 TI - How multiple interventions reduced injuries and costs in one plant. AB - Preventing work place injuries requires more than ergonomically correct work stations. In 1999, flexible plastic packaging producer, Pliant Corporation of Macedon, NY, initiated the presence of an onsite Physical Therapist to further efforts in making work stations safe and to develop a comprehensive approach in preventing and managing worker injuries. The process started with analysis of the functional demands of the work performed, elimination of ergonomic stressors, and development of daily stretching programs for employees to perform. In the second and third year of the program, objective functional job descriptions and minimum strength criteria were created as well as work task specific post-offer pre-work screens. This comprehensive approach has reduced the total number of OSHA recordable injuries, reduced the overall incident rate and assisted in reducing turnover. PMID- 16720965 TI - Occupational health systematic reviews: An overview. AB - Systematic reviews have been described by leaders in the field of evidence-based practice as the most powerful and useful evidence available to guide practice. This paper gives a description and analysis of systematic reviews currently included in the Cochrane Database for Systematic Reviews which have been identified by newly formed Cochrane Occupational Health Field as having an occupational health focus. PMID- 16720966 TI - The influence of a Functional Capacity Evaluation on fitness for work certificates in patients with non-specific chronic low back pain. AB - BACKGROUND: Comprehensive medical assessments in the majority of patients with chronic low back pain (CLBP) have failed to assess working capacity. Functional Capacity Evaluation (FCE) has become increasingly popular in the evaluation of working capacity in CLBP patients. This study investigates the influence of functional testing on decision making concerning medical fitness assessments for work. METHODS: A randomized clinical trial comparing the effect of a function centred (FCT) versus a pain-centred treatment (PCT) in patients with CLBP was performed. According to the legal requirements physicians issued Fitness for Work Certificates (FWCs) after rehabilitation. In the FCT group physicians were in possession of the results of FCE while this was not the case in the PCT group. Three experts assessed the FWCs and rated the quality and the information provided regarding working capacity in the previous work. RESULTS: Rating of the issued FWCs proved reliable (ICC = 0.77-0.92). Quality of FWCs and information regarding working capacity in the previous work differed significantly between the two groups (chi-square, p = 0.03, resp. = 0.008). There was a trend towards a higher attested work capacity in the FCT group (Mann-Whitney, p = 0.071). CONCLUSIONS: Functional Capacity Evaluation positively influences quality and information regarding working capacity of medical Fitness for Work Certificates in patients with chronic low back pain. PMID- 16720967 TI - Test-retest reliability of a WRULD functional capacity evaluation in healthy adults. AB - The aim of this study was to determine test-retest reliability of a Functional Capacity Evaluation for patients with non-specific Work Related Upper Limb Disorders (WRULD FCE). The study sample included 33 healthy adults, consisting of 14 males and 19 females with a mean age of 29.2 years. The WRULD FCE consisted of 8 different tests including 26 items measuring repetitive movements, duration, working in awkward positions, forceful movements and static postures. Two FCE sessions were held with a 10-day interval. Means, 95% confidence intervals, one way random Intraclass Correlation Coefficients (ICCs), 95% limits of agreement and repeated measures were calculated. An ICC between 0.75 and 0.90 was considered as good and an ICC above 0.90 was considered as an excellent reliability. The results showed that 14 of 26 items (54%) had excellent reliability, 9 of 26 items (35%) had good reliability and 3 of 26 items (11%) had moderate reliability based on ICC values. Significant learning effects were present in the Purdue Pegboard Task and in the Complete Minnesota Dexterity Test. It is concluded that the WRULD FCE is reliable in healthy adults. PMID- 16720968 TI - The functional IME: A linkage of expertise across the disability continuum. AB - Disability assessment remains a significant challenge especially in welfare systems like workers' compensation and disability insurance. Many of today's managed care strategies do not impact on the seminal issue of return to gainful employment. Employers, insurers, attorneys and case managers routinely request independent medical examinations (IMEs) as a means of determining degree of disability, functional limitations, work restrictions and "estimated" physical capacities. However, this approach is limited because physicians are not trained in the functional model of disability assessment. IMEs address pathology and impairments which represent a portion of the disability continuum described by the World Health Organization, Nagi, Guccione and others [e.g. pathology impairment-disability-handicap]. Functional capacity evaluations or FCEs are often performed by physical and occupational therapists who are trained in a function-based model of disability assessment. Unlike an IME physician who completes "Estimated Physical Capacities", therapists measure actual physical functioning. The value of both IMEs and FCEs can be enhanced through a "functional IME" that combines both models; medical-based examination and a function-based disability evaluation. This combination enhances the assessment of the relationship of pathology to impairment and impairment to disability status especially, in musculoskeletal disorders which tend to drive costs in workers' compensation. PMID- 16720969 TI - Use of Functional Capacity Evaluations by rehabilitation providers in NSW. AB - Returning the disabled worker to maximum productivity is a shared responsibility between the treating health practitioners and employers, with the insurer as an overseer of the injury management plan. Allied health professionals have responded to the need for standardized tools to assess work capacity by developing Functional Capacity Evaluations (FCEs). Literature exists outlining the limitations of FCEs in injury management systems; however, little is known about their uses and perceived usefulness. Questionnaires were posted to managers and therapists of accredited rehabilitation providers. The questionnaires consisted of both open and closed format questions. The response rate for the managers' questionnaire was 53%, while the therapists' questionnaire was 27%. The majority of rehabilitation providers use an assessment of their Own Design. Eight commercial FCEs were identified as being used. The majority of therapists use only one type of FCE. Therapists rated FCEs highly on their flexibility with Own Design assessment rated as significantly more flexible than the most frequently used commercial FCEs. From the data collected, it appears that cost is the main factor influencing choice of assessment. Level of evidence supporting capabilities of particular assessments does not appear to influence use. It seems that, despite there being poor research evidence to support the use of many brands of FCEs, they are still commonly used by rehabilitation providers. To achieve optimal use of FCEs, further research is required on the types of adaptations therapists make to commercial assessments and properties of Own Design assessments, including reliability and validity studies. PMID- 16720970 TI - Cultural diversity for health professions. AB - The purpose of this article is to help focus attention on why clinical health professionals need to be concerned about cultural diversity. It is not intended to be an exhaustive report on cultural diversity. Rather, attention is focused on the practitioner and how this provider can and should use and understand cultural diversity. While most of the examples in this article come from the press in the United States, the information has a much broader, universal and global implication. PMID- 16720971 TI - A review of employability and worksite interventions for persons with rheumatoid arthritis and osteoarthritis. AB - Arthritis has a significant effect on the US workforce. Significant economic effects and racial disparities have been found in treatment and health outcomes for persons with arthritis. This literature review focuses on the most commonly studied forms of arthritis, osteoarthritis (OA) and rheumatoid arthritis (RA), summarizing literature on employability trends, risk factors, and worksite health interventions for these conditions. Recommendations and future implications for research are given in relation to goals from Healthy People 2010. A brief description is provided of a worksite comparative study at the Missouri Arthritis Rehabilitation Research and Training Center (MARRTC), seeking to improve long term employability and functional outcomes for persons with arthritis. PMID- 16720972 TI - Work-related factors contributing to chronic disability in low back pain. AB - Low back pain is a costly and incapacitating musculoskeletal disorder. Prospective studies documenting the capacity of work-related factors to predict chronicity are few in number, the methodology used is very diversified, and the results obtained diverge. The aim of the present study is to investigate the capacity of work-related objective (non-psychosocial) and psychosocial factors to predict chronic disability related to low back pain. A longitudinal prospective study with two measurement times was carried out. The sample (N = 258) consisted of workers with subacute low back pain who were on sick leave and receiving compensation from the CSST (Quebec Workers' Compensation Board). Of all the work related variables measured, perceived stress and fears and beliefs about work were associated with return to work status at the six-month follow-up. The results obtained show the importance of considering fears and beliefs about work when identifying people in the subacute phase of low back pain who are at risk of developing chronic disability. PMID- 16720973 TI - Utilizing research in professional practice. AB - A review of the rehabilitation, disability studies, and allied health literature suggests the presence of numerous factors that impede the utilization of research by practitioners. The purpose of this article is to describe these factors and offer potential solutions that could enable rehabilitation professionals to integrate research findings into their practice. Specifically, the authors recommend strategies to (a) contextualize research throughout pre-service curricula, (b) establish researcher-stakeholder partnerships, (c) increase the accessibility of research to practitioners, and (d) establish professional and peer support for implementing research in field practice. PMID- 16720976 TI - Improving on nature: focusing the immune response on the V3 loop. AB - The conventional wisdom suggests that "constant" rather than "variable" regions of the HIV envelope (Env) glycoproteins would induce the most broadly reactive antibodies (Abs). However, of the several epitopes in the conserved regions of gp120 and gp41 that induce neutralizing Abs, all are well-protected by protein folding, glycosylation, and/or oligomerization of the Env proteins on the virus surface; most are only transiently exposed during the process of infection or are poorly immunogenic. In contrast, the third variable region (V3) of gp120 appears to be at least partially exposed during various stages of the infectious process, is immunogenic in essentially all HIV+ subjects, and is capable of inducing Abs able to neutralize a broad array of primary isolates. While these Abs were originally thought to be isolate-specific, a large body of data now shows that anti-V3 Abs from HIV-infected individuals indeed show intra- and inter-clade cross-reactivity with respect to both binding to diverse gp120 molecules and neutralization of many primary isolates. This cross-reactivity of anti-V3 Abs is counter-intuitive if one focuses on the sequence variability rather than on the conserved V3 structures which must be present in order to allow this region of the virus envelope to mediate selection of and interaction with chemokine receptors. Current data, summarized here, support the hypothesis that the V3 region of gp120 can induce broadly-reactive, cross-neutralizing Abs and as such should constitute a prominent target of the immune response induced with an HIV vaccine. PMID- 16720975 TI - Antibody polyspecificity and neutralization of HIV-1: a hypothesis. AB - HIV-1 has evolved many ways to evade protective host immune responses, thus creating a number of problems for HIV vaccine developers. In particular, durable, broadly specific neutralizing antibodies to HIV-1 have proved difficult to induce with current HIV-1 vaccine candidates. The recent observation that some broadly neutralizing anti-HIV-1 envelope monoclonal antibodies have polyspecific reactivities to host antigens have raised the hypothesis that one reason antibodies against some of the conserved HIV-1 envelope trimer neutralizing epitopes are not routinely made may be down-regulation of some specificities of anti-HIV-1 antibody producing B cells by host B cell tolerance mechanisms. PMID- 16720977 TI - Structural studies of human HIV-1 V3 antibodies. AB - X-ray crystallographic structures of two human, anti-V3 HIV-1 neutralizing antibodies, 447-52D and 2219, show how the two antibodies use different strategies for their cross-reactivity with different V3 sequences. 447-52D recognizes V3 primarily through main-chain hydrogen bonds to the N-terminal side of V3, with the GPGR tip region buried in the antigen-combining site. The side chains on the N-terminal side of V3 are exposed to solvent, allowing for sequence changes in this region, thus explaining how 447-52D can neutralize a wide array of viral isolates. Antibody 2219 contacts a more extensive V3 surface, with more side-chain involvement. However, residues at the GPGR tip are exposed to solvent, with no constraints on residue size, so that binding to unusual tip sequences with larger side chains such as RPRQ is possible. PMID- 16720978 TI - Ex vivo neutralization of HIV-1 quasi-species by a broadly reactive humanized monoclonal antibody KD-247. AB - By immunizing mice sequentially with six different V3 peptides we obtained a murine monoclonal antibody (MAb) C25, and its humanized counterpart KD-247. The MAb recognizes the sequence IGPGRA at the tip of the V3 loop and displays broad neutralizing activity against a variety of HIV-1 isolates. KD-247 was tested in an ex vivo neutralization assay to determine its capability to contain the spread of a quasi species population of clade B HIV-1 derived from two patients. The epitope of KD-247 was generally matching with the V3 sequences of various clones isolated from plasma and peripheral blood mononuclear cells (PBMC) of two patients. Complete or strong inhibition of viral replication was observed when the patients' PBMC were cultured with a high concentration of KD-247. Some neutralization escape variants, which had mutations in the V3 or outside of the V3 loop, emerged only at a low concentration of the MAb. These results suggest that KD-247 could be a good candidate for immunotherapy against HIV-1 in vivo. PMID- 16720979 TI - Neutralizing antibody patterns and viral escape in HIV-1 non-B subtype chronically infected treatment-naive individuals. AB - Here we studied the patterns of generation of neutralizing antibodies (NAbs) and virus escape during non-B subtype HIV-1 chronic infection among asymptomatic patients, and established whether a correlation exists between the generation of NAbs and the kinetics of CD4 T-cell decline. Therefore, sequential viruses and plasma obtained at 6 months to one year intervals over a three years period from ten HIV-1 group M subtype A, CRF02_AG, G, and H infected treatment-naive individuals were tested in neutralization assays. Overall, NAbs were present in all ten individuals, and had the capacity to neutralize autologous virus obtained six months earlier. Eight of the ten subjects showed an increasing capacity to neutralize early viruses and a low capacity to neutralize contemporaneous and later time-point viruses. The neutralizing activities within these individuals resulted in emergence of neutralization resistant viruses, and with the subsequent generation of more NAbs to the emerging resistant viruses. In the remaining two individuals, the capacity to neutralize early, contemporaneous, and later time-point viruses remained conserved. While the kinetics of CD4 T-cell decline varied among all ten individuals, there was no correlation with the capacity to generate NAbs in that, sequential plasmas from individuals with moderately or rapidly declining CD4 T-cells were capable of neutralizing early sequential viruses. We conclude from this study that in non-B subtype chronically infected asymptomatic patients with moderately and rapidly declining CD4 T-cells, potent NAbs are readily generated as the virus evolves to escape the effect of these antibodies. PMID- 16720981 TI - High frequencies of antibody responses to CD4 induced epitopes in HIV infected patients started on HAART during acute infection. AB - For many years only two human monoclonal antibodies (HMAbs) recognizing the CD4 induced (CD4i) epitopes of HIV-1 gp120 existed. Although a number of new CD4i HMAbs have been published recently, we have noted that in most attempts to produce HMAbs using EBV transformation a majority of antibodies produced in culture are lost within a few weeks. To determine what kinds of antibodies are made in these cultures we devised a semiquantitative culture to assess the frequency of B cells capable of producing antibodies and a microcompetition assay to determine what kinds of antibodies were made. Our results show that in three patients started on HAART during acute infection the most frequently produced antibodies binding to gp120 were directed against the CD4i epitopes. Our observations suggest that CD4i epitopes are much more immunogenic than had been previously appreciated. It is possible that envelope glycoproteins shed from virions and perhaps complexed with CD4 are responsible for eliciting these antibodies. The preservation of well regulated immune responses in these patients, together with repeated exposure to viral antigens (i.e. env), may explain the presence of larger than usual numbers of env-specific B cells that could be detected in EBV transformed cultures. PMID- 16720983 TI - Chloroquine-resistant Plasmodium falciparum local strain in Taiz Governorate, Republic of Yemen. AB - BACKGROUND: The emergence of Plasmodium falciparum resistance to widely used antimalarial drugs such as chloroquine has made malaria control and treatment much more difficult. In Yemen, 60% of the total population live in malarious areas. The problem of chloroquine resistance in Yemen is gradually worsening since the detection of first indigenous cases of P. falciparum resistance to chloroquine in 1989. METHODS: In a cross-sectional malariometric parasitic survey, 447 Yemeni children were enrolled from two selected districts (Hethran and Al-mafatch) representing Taiz Governorate. Duplicate thin and thick blood smears were prepared, stained with Giemsa stain and examined microscopically. Fifty-six students satisfied all criteria of the WHO for the assessment of P. falciparum response to chloroquine using a 7-day in vivo test. RESULTS: Out of 447 examined slides, 83 cases (100%) were found with falciparum malaria. The overall malaria parasite rate in Taiz Governorate was 18.6%, a prevalently mesoendemic condition. The obtained results of the 7-day in vivo study revealed that out of 83 P. falciparum cases who completed the study period, 56 cases did not respond to the standard dose of chloroquine, i.e. the overall resistance rate was 16.1%. The prevalence of chloroquine resistance was higher in the Hethran district (19.4%) compared with 10.0% in the Al-mafatch district. The majority had an RI resistance level. CONCLUSION: Chloroquine resistance of the local strain of P. falciparum was recorded in all studied districts in Taiz Governorate. This calls for an urgent revision of the current malaria treatment policy which still considers chloroquine as the first-line drug for treatment of uncomplicated P. falciparum malaria. To assess the magnitude of the problem, these districts could be the basis of future sentinel posts for continuous monitoring of chloroquine resistance in the whole country. PMID- 16720982 TI - Lack of long-term protective effect of antioxidant/anti-inflammatory therapy in transplant-induced ischemia/reperfusion injury. AB - BACKGROUND: Alloantigen-independent factors contribute to long-term damage in renal transplant recipients, likely due to ischemia/reperfusion (I/R) injury at transplantation (Tx). I/R injury promotes oxidative stress and inflammation resulting in endothelial injury. METHODS: In this study we investigated the long term efficacy (22 weeks) of short-term (10 day) endothelial protection therapy (EP) in 'optimal' donor kidneys using the male Fisher 344 rat isograft (ISO) model. ISO-EP kidneys were compared to untreated ISO (ISO-UN) kidneys. EP involved dexamethasone to donor, ex vivo treatment of the kidney with deferoxamine and tempol, and administration to the recipient of L-arginine and tempol for 10 days. Rats were sacrificed 22 weeks following Tx and compared to age-matched, normal controls. RESULTS: Both groups of ISO Tx rats developed similar renal dysfunction and structural damage and renal NADPH-oxidase-dependent O2- production was similarly elevated in ISO-UN and ISO-EP groups vs. controls. In vitro renal cortex NO synthase (NOS) activity was also similar in ISO-UN and ISO-EP rats, despite lower nNOS and eNOS protein abundance in ISO-EP. CONCLUSION: I/R injury-induced late graft dysfunction occurs even when optimal donors are used and when short-term EP treatment is given. Increased renal superoxide production is not prevented by short-term EP therapy. PMID- 16720980 TI - Characterizing anti-HIV monoclonal antibodies and immune sera by defining the mechanism of neutralization. AB - Understanding the nature of neutralization may provide information for crafting improvements in HIV vaccines. Using JR-FL as a prototype primary pseudovirus, we first investigated anti-HIV monoclonal antibodies (mAbs) in several neutralization formats designed to elucidate the timing of neutralization. MAb b12 was most effective before receptor binding, 2G12 neutralized effectively even after CD4 binding, and X5 and a V3 loop mAb (LE311) were inactive in a standard format but were induced by sCD4. Consistent with this latter finding, native PAGE indicated that X5 and V3 mAb binding to Envelope trimers was dependent on sCD4 binding. In contrast, 2F5 and 4E10 were active even post-CD4/CCR5 engagement. We next analyzed the neutralization mechanism of a panel of HIV+ donor plasmas of various potencies. All mediated high levels of post-CD4 neutralization that was not associated with activity in the standard format. None, however, neutralized effectively in the post-CD4/CCR5 format, suggesting that 2F5/4E10-like Abs were absent or at low concentrations. Finally, we analyzed a non-neutralizing plasma spiked with mAbs b12, 2G12 or 2F5, which resulted in increases in neutralization titers consistent with the activities of the mAbs. We conclude that these methods, together with other mapping approaches, may provide a better understanding of neutralization that could be useful in vaccine research. PMID- 16720985 TI - Factors affecting hemodialysis and peritoneal dialysis efficiency. AB - The peritoneal dialysis system has three major components: the peritoneal microcirculation, the peritoneal membrane, and the dialysate compartment. All these three that includes the composition of the solution and the modalities of delivery. All these three components may have an important impact on the final performance of the technique. As in the hemodialysis system, factors affecting diffusion of solutes as well as factors affecting convective transport may contribute to the final clearance of a given solute. Ultrafiltration responds to the same pressures applied to the extracorporeal dialysis system, but osmotic gradients represent by far the most important active component. PMID- 16720984 TI - Review of pharmacokinetic, pharmacodynamic and clinical studies with a modern combination of amoxicillin/sulbactam. AB - Amoxicillin/sulbactam is a modern antimicrobial combination. This combination proved to be useful for the treatment of several infections caused by different microorganisms, mainly with the beta-lactamase-producing species. In this review we present the most relevant pharmacokinetic, pharmacodynamic and clinical information associated with its use. PMID- 16720986 TI - Pathophysiology of peritoneal transport. AB - The peritoneal membrane has the surface area similar to the body surface area. It consists of mesothelial cells, interstitium, connective tissue fibers, blood vessels, and lymphatics. Solutes of various sizes traverse the peritoneal membrane through at least three various pores: 'large' pores located in the venular interendothelial gaps, small 'paracellular' pores, and ultrasmall, 'transcellular' pores or aquaporins localized in peritoneal capillaries and mesothelial cells. High molecular weight solutes are mass-transfer limited; thus, their clearances do not increase significantly with high dialysate flow. Clearances of small molecular weight solutes are dialysate flow limited. Ultrafiltration is proportional to the hydrostatic and osmotic transmembrane pressures. The peritoneum offers greater resistance to accompanying solutes than to water (solute sieving), so that the concentration of solutes in the ultrafiltrate is less than in plasma water. Sodium sieving leads to hypertension, which is frequently observed in patients treated with short-dwell or continuous flow peritoneal dialysis. Peritoneal equilibration test is the most commonly used test to characterize peritoneal function and select the most suitable dialysis technique for a patient. Long-term peritoneal dialysis is associated with progressive loss of ultrafiltration capability due to structural and functional alterations in the membrane mostly as the consequence of exposure to glucose degradation products or advanced glycation end products generated during the sterilization process. PMID- 16720987 TI - A simplified approach to understanding urea kinetics in peritoneal dialysis and hemodialysis. AB - Urea, a small molecular solute, is the candidate molecule commonly used to understand solute kinetics both in peritoneal dialysis and hemodialysis. Serum urea or serum urea nitrogen levels are utilized to calculate measures of dialysis adequacy and nutritional status in dialysis patients. The kinetic behavior of this molecule is however different for a continuous therapy like peritoneal dialysis vis a vis an intermittent therapy like hemodialysis. This article presents a simplified approach to understanding urea kinetics on dialysis. PMID- 16720988 TI - Peritoneal ultrafiltration: mechanisms and measures. AB - The net ultrafiltration during peritoneal dialysis equals the osmotic ultrafiltration minus the fluid loss. Fluid loss is made up primarily of hydrostatic pressure-driven convection to the tissues surrounding the peritoneal cavity and a lesser amount of lymph flow. The cellular and blood capillary sources of water are distributed in an interstitial matrix, which decreases the effective osmotic pressure in the vicinity of the microvasculature. The interstitial matrix makes the process of fluid removal far less efficient than it would be if the blood capillaries were actually in direct contact with the peritoneal fluid. Osmotically driven water transport occurs chiefly across the blood capillary endothelium into the interstitium and ultimately out into the peritoneal cavity. Aquaporin-1, water-only channels, are important for solute free water transport from blood capillaries. There may also be specialized water channels in cells, but this likely provides only a small component of solute-free water to the peritoneal cavity. Signs of ultrafiltration failure include loss of solute-free ultrafiltration and general decrease in ultrafiltration below 400 ml with a 4-h dwell of hypertonic dialysate. PMID- 16720989 TI - Evaluation of the peritoneal membrane function by three letter word acronyms: PET, PDC, SPA, PD-Adequest, POL: what to do? AB - This paper describes and reviews different methods to evaluate the peritoneal transport capacity. This evaluation is important because it will influence the preferred treatment regimen, and will also be a tool for longitudinal follow up both in the individual patient as in patient groups. PMID- 16720990 TI - The initiation of peritoneal dialysis: planning the initial prescription. AB - The initiation of peritoneal dialysis (PD) requires that the nephrologist address certain key issues. These include patient selection for PD, the timing of the placement of the PD catheter, the timing of the initiation of PD, the schedule of PD to be utilized, and the initial regimen of PD to be prescribed. In addressing these issues, the nephrologist needs to consider not only the medical condition of the patient but also the psychosocial status and quality of life concerns of the patient and his/her family. Furthermore, the dialysis treatment regimen needs to be organized to maintain the long term viability of PD therapy. PMID- 16720991 TI - PD membrane: biological responses to different PD fluids. AB - In chronic peritoneal dialysis (PD) alterations of both membrane structure and function occur with time on treatment, but also in relation to frequency and severity of infectious complications. In addition, therapy-associated factors such as the dialysis fluid bioincompatibility are important determinants for the longevity of the peritoneum as the dialysis membrane. Evidence available to date suggests that the elimination of glucose degradation products from multi chambered PD solutions is particularly important to improve peritoneal cell function as well as to reduce protein glycation and peritoneal advanced glycation end products deposition. Results from recent prospective randomized studies suggest that chronic treatment with these novel PD fluids may indeed lead to a significant improvement of clinical outcomes in chronic PD patients. PMID- 16720992 TI - Repopulation of the mesothelial monolayer during long-term experimental peritoneal dialysis. AB - BACKGROUND: Repopulation of the mesothelial monolayer after focal exfoliation, having the monolayer in vivo and in situ exposed to dialysis solutions, has not been thoroughly investigated. This study describes repopulation of a 'doughnut' like mesothelial ring exfoliated from the anterior liver surface of rats. METHODS: Animals were divided into 5 groups of 20 rats each. Group 1 - control unexposed animals: mesothelial cell imprints were taken after 1 (5 rats), 5 (5 rats), and 15 (10 rats) days following the procedure of exfoliation. Group 2 - sham injected animals. Group 3 - rats IP injected once a day, during 30 consecutive days with Hank's balanced salt solution. Groups 4 and 5 - same experimental protocol, but injecting 4.25% glucose single bag or 7.5% Icodextrin PDF. Imprints and/or biopsies were taken after a recovery period of 15 days, counted from the last IP injection. RESULTS: Density distribution of mesothelial cells in group 2 was not significantly different from that seen in unexposed rats, whereas that seen in group 3 Hank's balanced salt solution was marginally but significantly lower (p < 0.05) from that seen in controls. Eighty five percent of rats injected with 4.25% glucose developed fibrous adhesions, binding up together the exfoliated liver surface and the diaphragmatic muscle. For Icodextrin treated rats, the prevalence of fibrous adhesions was 95%. CONCLUSIONS: The 'doughnut' experimental model appears as a promising tool for in vivo and in situ investigation of mesothelial repopulation. Both osmotic agents substantially restrain mesothelial repopulation, leading to repair by connective tissue. PMID- 16720993 TI - Fibrosis and sclerosis: different disorders or different stages? AB - Peritoneal sclerosis is very common in peritoneal dialysis (PD) patients. It can vary from the mild, clinically silent sclerosis always present after years of PD, to rare but dramatic and often fatal cases. In our opinion, peritoneal sclerosis is a single disorder, so its variable manifestations are different stages of one nosological entity: this opinion relies mainly on strong connections in pathophysiology. In our view, the frequency, pathology, animal models, etiology and pathogenesis often show a bimodal configuration with suggests that peritoneal sclerosis is actually two distinct nosological entities: simple sclerosis and sclerosing peritonitis. The former is very frequent, with minor anatomical alterations and low clinical impact; it is reproducible in animals by means of PD, and is clearly due to the poor biocompatibility of PD. The latter is rare, with radical anatomical alterations and high mortality; it can only be reproduced in animal models by means other than PD and seems to be due to factors both related and unrelated to PD. PMID- 16720994 TI - What did we learn from animal models in peritoneal dialysis? AB - The development of animal models in peritoneal dialysis has led to some breakthroughs in the application of this dialysis modality in clinical practice. These breakthroughs are (1) a better understanding of the physiology and pathophysiology of solute transport and ultrafiltration mechanisms, (2) the observation and integration of the long-term structural and functional alterations of the membrane, (3) a better understanding of the biocompatibility issues involved in PD, leading to the clinical introduction of more biocompatible dialysis solutions and finally, (4) the development of colloid osmotic solutions containing polyglucose polymers for application in the long dwells. Intravital miscroscopy provides information in live animals about diverse functional parameters, such as blood flow rate, vessel diameter, permeability to macromolecules, leukocyte-endothelium interaction, capillary recruitment, and lymph vessel kinetics. Also evaluation of different parameters in a living experimental animal, allowing integration of function and structure is possible. A variety of chronic PD models have been developed, mainly to study effects of long-term peritoneal dialysate exposure on peritoneal membrane function and structure. The implementation of different blocking agents of biochemical substances in these models has elucidated many molecular biological mechanisms involved in these processes. The important roles of aquaporins, vascular endothelial growth factor, nitric oxide, advanced glycation end product formation and their receptor (RAGE) upregulation and the integrated roles of all these factors in the fibrotic alterations of the membrane as observed in patients on long-term PD have been investigated. More recently, genetically modified mice have been used as an important tool to investigate the molecular basis of peritoneal changes during dialysis and during acute peritonitis. PMID- 16720995 TI - GDP and AGE receptors: mechanisms of peritoneal damage. AB - Long-term peritoneal dialysis (PD) is limited by morphological changes of the peritoneal membrane. Structural changes were promoted by toxicity of glucose degradation products (GDPs) which are generated during heat sterilization in peritoneal dialysis fluids (PDFs). Besides their direct toxicity GDPs promote formation of advanced glycation endproducts (AGEs). RAGE (receptor for AGE) is the best characterized signal transduction receptor for AGEs and is expressed on mesothelial cells. The effects of PDFs with different amounts of GDPs were compared on morphological changes in the peritoneal membrane in a RAGE -/- mouse model. It could be demonstrated that RAGE plays a pivotal role in structural damage (e.g. inflammation, neoangiogenesis and fibrosis) of the peritoneal membrane. Further investigations of this pathway with regard to preventing peritoneal fibrosis should be performed to maintain the integrity of the peritoneal membrane in peritoneal dialysis patients. PMID- 16720996 TI - Effects of peritoneal dialysis on the vascular bed of peritoneal membrane. AB - The vascularization of the peritoneal membrane is a key factor in the effectiveness of peritoneal dialysis as a dialysis modality. Over the last years, our understanding of how the peritoneal vasculature reacts to instillation of peritoneal dialysate has substantially improved. The changes in the structure of the peritoneal function over time on peritoneal dialysis as found in functional tests has been confirmed in biopsy studies performed on patients. These show both neoangiogenesis and fibrosis as the underlying morphological changes contributing to these phenomena. A thorough understanding of the biological processes involved in these alterations will be the clue for improvement of the treatment and a key to develop strategies to avoid these deformations. PMID- 16720997 TI - Effect of PD solutions on patient outcome. AB - With the recognition of the bio-incompatibility of conventional glucose based peritoneal dialysate (PDF), many new PDF has been developed and commercially available. All of them contain much less low glucose degradation product (GDP) solutions. Additional features are neutral or physiological pH, or glucose replaced by amino acid or glucose polymer icodextrin. These fluid has been shown to be more biocompatible in in-vitro and animal studies. There are short term randomized studies showing some benefit in certain clinical criteria, like increase in CA 125 in effluent PDF, better preservation of residual renal function, nutritional status, fluid and cardiovascular status, but beneficial effect on patient survival and long term peritoneal function has not been demonstrated apart from a non-randomized cohort study. To show such benefit on patient outcome, long term prospective randomized study is needed. PMID- 16720998 TI - Peritoneal transport with icodextrin solution. AB - Icodextrin is the only large molecular weight osmotic agent available in peritoneal dialysis solutions. Icodextrin (compared to glucose) has a prolonged positive ultrafiltration because of the slow absortion of icodextrin due to its large molecular weight. As icodextrin induces transcapillary ultrafiltration by a mechanism resembling 'colloid' osmosis (with the ultrafiltration occurring mainly through the small pores) almost no sieving of solutes is observed with icodextrin based solution resulting in increased convective transport and clearance of small solutes. In general, the transport characteristics of the peritoneal membrane are similar with glucose- and icodextrin-based solution, but results from the EAPOS study suggests that use of icodextrin-based solution may be associated with less changes in peritoneal transport with time. PMID- 16720999 TI - Dry body weight: water and sodium removal targets in PD. AB - BACKGROUND/AIMS: Cardiovascular mortality is high in peritoneal dialysis patients. This may be due to the presence of hypertension and fluid overload. Dietary intake of water and sodium are likely to be important, especially in anuric patients. METHODS: A review of the literature on assessment of fluid status, and on peritoneal removal of water and sodium. RESULTS: MF-BIA added to clinical judgement is helpful for the assessment of fluid status. Natriuretic hormones in patients without heart failure are promising, but their value in clinical practice has not been established yet. Peritoneal ultrafiltration and sodium removal in anuric peritoneal dialysis patients are associated with survival. APD patients have lower sodium removal than CAPD, icodextrin increases it. The value of routine use of low sodium dialysis solutions has not been established. Definite targets on fluid and sodium removal have not been established, but the maintenance of euvalemia, while reducing the exposure to 3.86% glucose solutions at the same time, is largely facilitated by dietary salt and water restrictions. CONCLUSION: MF-BIA is a useful addition to clinical judgement of hydration status. The use of high dose loop diuretics facilitates water and salt removal in the presence of residual renal function. A dietary salt restriction of 4 g/day in anuric patients improves hypertension and is likely to reduce fluid intake. PMID- 16721000 TI - Is there a competition between urine volume and peritoneal ultrafiltration in peritoneal dialysis patients? AB - Many peritoneal dialysis patients are overhydrated. Overhydration may lead to hypertension and left ventricular hypertrophy, and may be related to inflammation and malnutrition. The presence of overhydration is not always detected by clinical examination. Especially patients with peritoneal ultrafiltration failure and/or negligible residual renal function are prone to overhydration. Treatment consists of a combination of sodium and fluid restriction, in combination with increased peritoneal ultrafiltration and loop diuretics in patients with residual diuresis. Peritoneal ultrafiltration can be enhanced by the use of hypertonic glucose solutions or icodextrin. In some, but not all, studies, residual renal function or diuresis declined with an increase in peritoneal ultrafiltration. At least in a subset of patients, underhydration might have played a role in this phenomenon. We propose to treat overhydration using both clinical criteria and objective techniques such as vena cava echography or bioimpedance measurements. PMID- 16721001 TI - Application of bioimpedance techniques to peritoneal dialysis. AB - Peritoneal dialysis (PD) has been used as a home dialysis therapy for renal replacement for more than 30 years. In a recent assessment of treatment quality, the mortality of patients on PD was referenced as being higher than of those on hemodialysis. Several reports suggest that a high proportion of PD patients are overhydrated. Clinical assessment of dry weight in PD patients is difficult and further complicated by the paucity of signs and symptoms indicative of dehydration (such as intradialytic hypotension or muscle cramps). Monitoring tools used for fluid status estimation during hemodialysis, e.g. online blood volume and blood pressure measurement, are not readily available in PD patients. Bioimpedance analysis technique has been considered as a potential tool to measure body fluid non-invasively, inexpensively and simply. Although Bioimpedance analysis has been used in clinical studies for more than 20 years, the knowledge of the electrical properties of body tissues is still evolving. In this review we aim to clarify the principles of different bioimpedance techniques and to introduce their applications in PD patients. PMID- 16721002 TI - Peritoneal dialysis: a clinical update congestive heart failure and PD. AB - BACKGROUND: There is a belief that peritoneal dialysis may be an important treatment modality for refractory heart failure, allowing at least an improvement in quality if not quantity of life during the last stage of this debilitating chronic disease. This paper examines the rationale behind this modality, critically appraises the available literature, calls for more research in the area and puts forward a framework for considering peritoneal dialysis in refractory heart failure. CONCLUSION: When faced with a patient with refractory heart failure admitted to hospital on multiple occasions because of complications of volume overload, the following approach to initiating peritoneal dialysis should be considered: subjects for dialysis must have a minimal blood pressure, and those whose hemodynamic status improves with diuresis, even though they develop a pre-renal picture, may be the best candidates. PMID- 16721003 TI - MIA syndrome in peritoneal dialysis: prevention and treatment. AB - Malnutrition, inflammation and atherosclerosis are prevalent in end stage renal disease and constitute the Malnutrition-Inflammation-Atherosclerosis Syndrome. The syndrome is associated with high cardiovascular mortality and accounts for most of the premature deaths in peritoneal dialysis patients. Presence of elevated C-reactive protein levels correlates with malnutrition, decreased fluid removal and mortality in these patients. Early recognition of the syndrome is important to identify high risk patients. Nutritional support, changes in dialysis and drug therapy may decrease the cardiovascular morbidity and mortality. PMID- 16721004 TI - Being an inflamed peritoneal dialysis patient - a Dante's journey. AB - Patients on peritoneal dialysis (PD) suffer from a high incidence of cardiovascular disease as compared to healthy individuals, and a markedly increased mortality that cannot fully be explained by traditional risk factors. Recent evidence suggests that end-stage renal disease is a state of systemic inflammation and oxidative stress, both of which appear to play an important role in the development of cardiovascular disease. Here, we review recent data looking at the impact of persistent inflammation (usually recognized by elevated serum levels of C-reactive protein) on morbidity and mortality in PD-patients. While many causes of inflammation are attenuated with PD, including volume overload and biocompatibility of membranes, PD is also associated with modality-specific causes of inflammation such as peritonitis, glucose degradation products and fluid bioincompatibility. Additionally, PD can lead to the uptake of large amounts of glucose from the fluid, resulting in a risk of increased oxidative stress, which also may contribute to inflammation. In addition, recent research shows that genetic factors are clinically important in determining inflammatory response. Finally, we briefly explore potential strategies specifically aiming at reducing intraperitoneal and systemic inflammation in PD patients. PMID- 16721005 TI - Serum parameters, inflammation, renal function and patient outcome. AB - Early serum parameters predicting patient outcome mainly are albumin and prealbumin. They were initially thought to represent nutritional status. With the understanding of the contribution of inflammation to malnutrition, cardiovascular disease and mortality, these negative phase inflammatory markers probably represent inflammation more than nutrition. Similarly, C-reactive protein is also found to be a strong predictor of mortality. The other strong predictor of patient outcome is residual renal function. The interlink between preservation of residual renal function and inflammation is becoming a subject of growing interest. PMID- 16721006 TI - Sterile solutions and peritoneal inflammation. AB - There is now abundant evidence in both animal and human studies that sterile dialysis solutions result in the alteration of the peritoneal barrier. Mesothelial cells undergo an epithelialto- mesenchymal transition after less than 1 year of dialysis. After more than 6 years of peritoneal dialysis, there is extensive fibrosis and neoangiogenesis in the sub-mesothelial compact zone. Clinical studies demonstrate that the structural changes correlate with alterations in transport function and progressive ultrafiltration failure. The mechanisms of inflammation include cellular peroxide production, acidic solutions, glucose and its degradation products, and the integrated response of the cellular-cytokine cascade. Basic and translational research efforts are discussed to highlight our current knowledge in this area and the remaining questions. PMID- 16721007 TI - Adipokine signaling in the peritoneal dialysis patient. AB - Patients on peritoneal dialysis suffer from an increased incidence of cardiovascular disease, obesity, insulin resistance, as well as a markedly increased mortality rate. In both the general population and in renal patients current research implicates fat tissue as an important factor in modulating many of these complications through the secretion of various signaling peptides, mainly cytokines and adipokines. Whereas hemodialysis patients tend to lose fat mass after initiation of renal replacement therapy, peritoneal dialysis-patients instead gain fat, probably as a result of continuous glucose absorption. In the present overview we summarize some recent findings indicating that fat mass actively contributes to systemic inflammation and metabolic disturbances in chronic kidney disease. We conclude that adipokines are likely to influence several key survival factors in peritoneal dialysis patients, including systemic inflammation, endothelial health and appetite. PMID- 16721008 TI - The effect of fibrosis on peritoneal transport. AB - Current sterile dialysis solutions cause inflammation in the sub-mesothelium that lead to fibrosis, angiogenesis, and eventual ultrafiltration failure. While the normal interstitium separates the peritoneal microvasculature from the dialysis fluid and makes trans-peritoneal transport less efficient, changes in the sub mesothelial layer can result in progressive increases in solute transfer and ultrafiltration diminution. Years of exposure to dialysis solutions result in an epithelial-to-mesenchymal transition of mesothelial cells and the formation of an avascular layer of intersitial matrix and plasma proteins in the sub-mesothelial compact zone. The formation of this fibrotic layer separates the dialyzing solution from the exchange microvessels and markedly decreases the effective osmotic pressure near the exchange microvessels. Angiogenesis dissipates the osmotic driving force through increases the perfused vascular area and marked increases in solute permeability. These combined changes result in more rapid loss of the osmotic agent and diminished ultrafiltration. PMID- 16721009 TI - Peritoneal dialysis infections recommendations. AB - Peritonitis remains a serious problem in peritoneal dialysis patients accounting for technique failure and contributing to mortality. Many peritonitis episodes are due to contamination at the time of the exchange and exit site infections. Protocols can be implemented by programs to diminish the risk of infection. Careful training, especially in handwashing technique and in doing the connection, are critical for preventing contamination related peritonitis. Peritonitis due to exit site infections can be reduced by use of exit site antibiotic cream. Gentamicin as opposed to mupirocin exit site prophylaxis reduces not only S. aureus but also P. aeruginosa infections. Refractory exit site infections can be managed with simultaneous catheter replacement. Once peritonitis occurs, prompt institution of empiric antibiotics, dictated by the history of the program's infections, should be done. Initial therapy is then modified once the culture results are known. Catheters require removal if the peritonitis fails to resolve within 5 days of appropriate antibiotic therapy or if peritonitis is relapsing. Fungal peritonitis is best treated with prompt catheter removal. Implementation of protocols to prevent peritonitis and careful attention to both the organisms causing peritonitis and the rate of infection by a peritoneal dialysis center are essential for reducing infectious complications. Once infections occur, rapid steps to treat and manage are important to diminish the risk of mortality and subsequent peritoneal damage, areas requiring more research. PMID- 16721010 TI - Cloudy peritoneal dialysate: it's not always infection. AB - BACKGROUND/AIMS: Turbid peritoneal dialysate is most commonly due to bacterial peritonitis. However, not all instances of cloudy dialysate are due to infection. This paper will review the various non-infectious causes of cloudy dialysate fluid. METHODS: Literature review and synthesis. RESULTS: Cloudy dialysate may be due to pathologic increases of either cellular or non-cellular constituents of peritoneal fluid. Polymorphonuclear leukocytes may be increased due to either intra- or juxtaperitoneal inflammation or drug-induced chemical peritonitis. Increased eosinophils often represent a response to intraperitoneal air or an allergy to a component of the dialysis system. Red blood cells may be present due to one of many causes. Monocytes or malignant cells are relatively uncommon. The differential for the non-cellular causes of culture-negative cloudy dialysate is limited to elevated fibrin or triglycerides. The latter may be due to lymphatic obstruction, pancreatitis, catheter trauma, dihydropyridine calcium channel blockers, or the superior vena cava syndrome. CONCLUSION: After ruling out atypical infectious etiologies, a diverse set of aseptic causes remains in the differential diagnosis. Use of an organizational scheme based upon identification of the cellular or non-cellular constituent producing dialysate turbidity facilitates appropriate diagnostic and therapeutic interventions. PMID- 16721011 TI - Peritoneal access: the past, present, and the future. AB - In the early years of peritoneal dialysis, there was no specific device for peritoneal dialysis; rather the devices used in general surgery and urology were taken advantage of for peritoneal access. These early devices, used for short term peritoneal dialysis, were plagued with multiple complications, such as pressure on intestines of rigid tubes, plugging of openings, leakage of fluid around the access, and difficulties in fixation of the tube on the abdominal wall. In the late 1940s, after World War II, multiple peritoneal dialysis solution compositions and multiple peritoneal accesses were tried, and first accesses specifically for peritoneal dialysis were designed. In the 1950s and particularly 1960s new access features solved most of the problems and eliminated most complications of peritoneal dialysis performed in the supine position. The invention of silicone rubber catheter with polyester cuff(s) was a greatest breakthrough in peritoneal dialysis access development. Unfortunately, none of the currently used catheters is trouble free; poor dialysate drainage, pericatheter leaks, exit site and tunnel infections, and recurrent peritonitis episodes are frequently encountered. Therefore, there is an incessant search for new technological solutions, including new shapes of intraperitoneal and intramural catheter segments, and new catheter materials are tried. PMID- 16721012 TI - How to optimise anaemia therapy in peritoneal dialysis patients. AB - As with other groups of chronic kidney disease patients, the treatment of anaemia is of paramount importance in the general management of patients receiving regular peritoneal dialysis. The availability of agents able to stimulate erythropoiesis has transformed the management of anaemia in CKD, but questions are still raised as to the optimum means of using these drugs. Iron management is also pivotal to the satisfactory correction of anaemia, and again there is much discussion as to whether oral or intravenous iron is the preferred mode of administration in peritoneal dialysis patients. On the basis of the published evidence to date, PD patients should maintain a haemoglobin above 11 g/dL in line with the US and the European Anaemia guidelines, and intravenous iron should be used to correct any iron deficiency. Oral iron may be effective in a minority of patients. This article aims to explore some of these issues in greater detail so that patients on peritoneal dialysis can derive the greatest benefits from correction of anaemia and maintenance of an adequate haemoglobin. PMID- 16721013 TI - Calcium and phosphate handling in peritoneal dialysis. AB - In the last 10 years, it has been well documented that mineral metabolism abnormalities in dialysis patients are associated with an enhanced risk of morbidity and mortality for cardiovascular disease. Extraskeletal calcifications represent one of the major risk factors involved in the pathogenesis of cardiovascular disease in this population. In fact, secondary hyperparathyroidism and hyperphosphatemia associate with increased cardiovascular mortality in uremic patients for two reasons: first for the passive deposition of calcium and phosphate in soft tissues; second for the active role of inorganic phosphate on direct induction of extraskeletal mineralization of the tunica media in the vasculature of these patients. In peritoneal dialysis patients, many unbalances of calcium and phosphate metabolism are present. In particular, recent cohort studies indicate that most patients do not reach targets indicated by clinical practice guidelines. Further efforts to control hyperphosphatemia are essential, in order to reduce the impact of secondary hyperparathyroidism both on bone and cardiovascular system. PMID- 16721014 TI - Integration of peritoneal dialysis in the treatment of uremia. AB - The real integration of a specific therapy into the renal replacement program is represented by the possibility of easy and free patient transfer from one treatment to another without restrictions. In the case of peritoneal dialysis we feel that its integration in the therapeutic approach of uremia represents an ethical obligation for the physician, a clinical opportunity for the patient and a good cost/benefit solution for care givers. A full conviction that peritoneal dialysis represents a real therapeutic option for ESRD patients is necessary to achieve a real integration of this therapy in the uremia treatment program. A positive cost benefit ratio, both from the clinical and the economical points of view must also be seeked. The patient indirectly, must receive the same positive conviction, based on solid data and clinical results, comparable to those achievable in hemodialysis. Furthermore the patient must know that such treatment will provide an equal opportunity for kidney transplant compared to other therapies. Such a kind of feeling and knowledge must include information on patient's survival, rate of complications, treatment adequacy, availability of different techniques within the treatment and complete summary of advantages and disadvantages. PMID- 16721015 TI - The role of chronic peritoneal dialysis in the management of the patient with chronic kidney disease. AB - The role of chronic peritoneal dialysis (CPD) in the management of patient with chronic kidney disease is being reexamined. There has been a decline in CPD utilization in the United States, which has been attributed to a variety of factors. It is important that these factors be understood and addressed, since nephrologists feel that about 30% of end-stage renal disease patients should be maintained on CPD and patients are more satisfied with CPD and feel that CPD has a less negative impact on their lives than hemodialysis. The present chapter discusses some of the reasons for decline in CPD utilization in the United States. PMID- 16721016 TI - Peritoneal dialysis is appropriate for elderly patients. AB - BACKGROUND/AIMS: The utilization of peritoneal dialysis decreases with age. A number of concerns have been raised regarding the suitability of peritoneal dialysis for elderly patients. The purpose of this review is to determine whether these concerns are medically valid. METHODS: Literature review and synthesis. RESULTS: Most elderly patients possess the manual and cognitive skills necessary to perform peritoneal dialysis. Elderly patients on peritoneal dialysis exhibit excellent compliance with their treatment regimen and display no increase in the rate of infectious complications though they may have a slight increase in hospital days. They easily achieve adequacy targets, experience good technique survival and their nutritional status is at least as good as that of their hemodialysis counterparts. Patient survival varies around the world but is overall comparable to that of age-matched patients on hemodialysis. Quality of life may be somewhat superior to that of older hemodialysis patients. CONCLUSION: Elderly patients with end-stage renal disease are appropriate candidates for peritoneal dialysis. It is not medically justifiable to exclude them from consideration for this therapeutic modality. PMID- 16721017 TI - The impact of peritoneal dialysis upon quality of life and mortality of patients with end-stage congestive heart failure. AB - BACKGROUND: Aged patients affected by congestive heart failure refractory to pharmacologic therapy, coupled with severe comorbidities that preclude heart transplantation have a gloomy prognosis. Use of automated peritoneal dialysis resulted in a significantly improved quality of life and survival. METHODS: Twenty patients (mean age: 65.7 +/- 7.7 years) with severe congestive heart failure (Class IV-NYHA) and a high Charlson comorbidity index (7.8 +/- 1.8) were treated by means of automated peritoneal dialysis (3 sessions/week, 8 h each). The period of follow-up ranged between 7 and 35 months (19.8 +/- 7.4 months). RESULTS: After 1 year of follow-up, all patients showed functional (Class I-NYHA) and hemodynamic improvement: left cardiac work index increased from 2.33 +/- 0.69 to 2.59 +/- 0.47 kg/min/m(2) (p < 0.01); systolic times ratio was significantly reduced (from 61.14 +/- 12.57 to 39.18 +/- 13.44%, p < 0.01); and the thoracic fluid contents was substantially reduced (from 0.04 +/- 0.005 to 0.003 +/- 0.0001 Omega). A marked drop of hospitalization days was observed. CONCLUSIONS: Automated peritoneal dialysis appears as a promising tool for treating patients suffering from refractory, end stage congestive heart failure. Improvement of cardiac function may result from clearing the blood from middle molecular weight myocardial depressant substances, including atrial natriuretic peptide. Results of this investigation are a call for a prospective multicenter study in order to confirm these promising observations. PMID- 16721018 TI - The impact of the pre-transplant renal replacement modality on outcome after cadaveric kidney transplantation: the ghent experience. AB - For suitable patients, renal transplantation is still the most preferable renal replacement modality, offering the best outcome in terms of survival and quality of life [Meier-Kriesche, H.U. et al: Semin Dial 2005;18:499-504]. The shorter the period on dialysis, the better the outcome after transplantation seems to be [Meier-Kriesche, H.U. et al: Transplantation 2002;74: 1377-1381]. However, for most patients, a pre-emptive transplantation is not an option by lack of a suitable organ. Therefore, most people have to undergo hemodialysis or peritoneal dialysis (PD) while awaiting a donor kidney. There is evidence that PD positively impacts on the outcome after transplantation [Van Loo, A.A. et al: J Am Soc Nephrol 1998;9:473-481], an effect that could be attributed to a more stable fluid homeostasis, but also to an independent effect of biocompatibility of the dialysis membrane [Van Biesen, W. et al: Transplantation 2000;69:508-514], which is by definition better in PD. Based on these findings, since 1999, all hemodialysis patients at the university of Ghent are dialyzed on a low complement activating dialyzer, and dialysis and especially ultrafiltration in the 24 h preceding the transplantation are avoided as much as possible. A recent re analysis of the data of the outcome of our transplant program showed that this approach resulted in a reduction of delayed graft function in the hemodialysis patients, allowing to reach an outcome level comparable to that of the PD patients. However, the long-term patient survival still is slightly superior in the PD patients. PMID- 16721020 TI - Peritoneal dialysis after a failed transplant. AB - Failed transplantation is an increasingly common cause for starting dialysis treatment. As with all patients approaching dialysis there is a need for adequate physical and psychological preparation and yet whilst by definition these individuals are known to health professionals this is not always achieved. It is likely that given adequate information, a significant proportion of these patients would prefer PD on lifestyle grounds. There is increasingly strong evidence that patients commencing PD after transplant failure enjoy overall survival and technique survival that is no different to those new to dialysis, even when other risk factors such as age, comorbidity, race, gender and membrane function are taken into account. The risk of peritonitis is also not different. These patients tend to lose residual renal function more rapidly but this does not translate into worse outcomes. The role and benefit in modulating immune suppressive drugs before and after commencing PD is not clear. PMID- 16721019 TI - Transplantation outcome in patients on PD and HD. AB - In the past, peritoneal dialysis (PD) has been considered a second choice dialysis modality for many aspects and that negative attitude has been extended also to possible negative effects on renal transplantation. In the last years, many papers have faced the question whether PD could attain similar results in renal transplantation as hemodialysis and there is sufficient evidence to answer that question. On the short time after transplantation, patients coming PD have lower prevalence of delayed graft function than hemodialysis patients, but higher prevalence of renal vascular thrombosis, above all in children. Incidence of acute graft rejection is not different between the two dialysis modalities. The long-term outcome of renal transplantation is similar in patients coming from either PD or hemodialysis. PMID- 16721021 TI - Automated peritoneal dialysis - indications and management. AB - Automated peritoneal dialysis (APD) use has increased considerably in the last decade, and its growth has been mainly driven by patient preference and development of new, simpler cyclers. Careful management of the APD prescription can result in adequate solute and fluid removal in vast majority of patients, even anuric and large patients. So far, there are no convincing data that peritonitis rates, decline in residual renal function, protein losses, patient and technique survival, are different for APD and continuous ambulatory peritoneal dialysis (CAPD). APD has the major advantage of allowing positive changes in the life-style of end stage renal disease patients. PMID- 16721022 TI - APD schedules and clinical results. AB - This chapter exposes the clinical effects of different APD schedules. Clinical studies have shown that Na removal is lower with APD compared to CAPD. Therefore the loss of residual renal function requires continuous therapy with long day dwell of polyglucose dialysate. Peritoneal small molecule clearances are closely determined by the hourly dialysate flow rate (and not the dwell time, which is a concept coming from equilibrium PD techniques as CAPD) with a maximum reached by 3 l/h for average transporter patients. On the other hand, the optimal intraperitoneal volume should be 1500 ml/m(2) of BSA, and less if the hydrostatic intraperitoneal pressure is higher than 18 cm H(2)O. The optimization of the nocturnal APD session depends on the knowledge of the individual drain flow profile. A new schedule, called 'BreakPoint-APD' is based on the automatic adaptation to the drain profile for each patient and for each cycle. It increases clearances by about 10% compared to tidal and CCPD, though reducing the number of nocturnal alarms. The future of APD is likely to continue thanks to further simplification in the machine use, i.e. with improvements in cycler technology. PMID- 16721023 TI - Evolution of technology for automated peritoneal dialysis. AB - Automated peritoneal dialysis (APD) is important for the further penetration of PD in the dialysis marketplace. Long dwell, equilibration PD (CAPD) has limited applicability in many patients due to inadequate solute clearance or fast membrane transport characteristics. Providing large volumes of dialysate over circumscribed hours is highly labor intensive without an automated system. Early attempts at APD were crude but effective in reducing labor, which was generally provided by nursing staff. Later evolution of PD technology has been greatly accelerated by the microchip, and by miniaturization of components. Current generation machines allow individualized fill volumes, variable tidal volumes and additional daytime automated exchanges, teledialysis, memorized delivery control, and full portability. The ideal machine should not only be able to perform all treatment schedules, but it should also optimize the performance of a selected treatment strategy. Biocompatible solutions, improved osmotic agents, and sorbent technology are all adaptable to APD. The eventual evolution toward continuous flow PD will resolve many of the current problems with both CAPD and APD. PMID- 16721024 TI - Continuous flow peritoneal dialysis: current state-of-the-art and obstacles to further development. AB - Peritoneal dialysis (PD) is still underutilized as home based renal replacement therapy and in-patient treatment of acute renal failure. Hindering the expansion of PD is poor solute clearance, which is a result of the intermittent dwell technique. Continuous flow PD is an old concept that has demonstrated urea clearances from 2-5 times higher than standard PD. It relies on a 2-3 l dwell volume and continuous dialysate flow at 100-300 ml/min. This high flow rate dictates the need for an efficient dual lumen catheter, or two separate catheters with ports separated maximally, as well as a means to generate or regenerate large volumes of fluid. A modified hemodialysis system can easily be adapted to regenerate sterile peritoneal dialysate, and a dual lumen catheter with excellent flow characteristics has been designed. Ultrafiltration control and a means to accurately balance transperitoneal with external ultrafiltration persist as technical challenges. Continuous flow PD remains an attractive modality for daily home dialysis and treatment of acute renal failure. PMID- 16721025 TI - Continuous flow peritoneal dialysis: ideal peritoneal dialysis or second-rate hemodialysis? AB - BACKGROUND: Since the original description of continuous flow PD (CFPD) by Shinaberger, there has been interest in the utility of this modality to augment small solute clearances in those on PD. METHODS/RESULTS: The clinical and theoretical treatment of CFPD regimens and the changing views of small solute clearance and outcome in dialysis since the publication of several important randomized, controlled trials are reviewed. CONCLUSION: There is little justification to promote CFPD solely on the basis of increased small solute clearance, given the lack of robust association between this measure and outcome in PD patients. Furthermore, leaving the abdomen empty between CFPD sessions may seriously compromise removal of middle molecular weight uremic toxins, one of the presumed strengths of conventional PD. PMID- 16721026 TI - A new home based bioimpedance system for PD. AB - Fluid overload and uncontrolled hypertension may be considered important mortality risk factors in peritoneal dialysis (PD) population. Even malnutrition is highly prevalent in PD patients. It is now well established that lower levels of serum markers of nutrition such as albumin, creatinine, and prealbumin are associated with increased mortality in PD patients [Fein, P.A. et al: Adv Perit Dial 2002;18:195-199]. Moreover cardiovascular disease is a leading cause of death in patients with end-stage renal disease, and hypertension and volume expansion are highly prevalent in long-term PD patients. Many studies in hemodialysis and in PD have demonstrated that phase sensitive bioelectrical impedance analysis is a widely used and proven method for evaluating patient's body composition. The vectorial bioimpedance analysis is a validated system to evaluate the hydration and nutritional state of hemodialysis and PD patients with acceptable sensitivity and specificity. The aim of this study is to evaluate the reliability and accuracy of the new multifrequency BodyComp bioimpedance analyzer as a home based tool versus traditional Bia Vector. PMID- 16721027 TI - The potential application of sorbents in peritoneal dialysis. AB - Sorbents have been designed to remove small and middle molecular weight toxins, including low molecular weight proteins, peptides, cytokines and chemokines in patients undergoing renal replacement therapy. Sorbents assist the process of peritoneal dialysate regeneration and have the potential to improve the efficiency of continuous flow peritoneal dialysis. PMID- 16721028 TI - Role of node dissection for lymphatic remetastasis in repeat hepatectomy for colorectal liver metastasis. AB - BACKGROUND: Although the prognosis after hepatectomy for colorectal liver metastasis with hilar node remetastasis is poor, the role of node dissection for lymphatic remetastasis at repeat hepatectomy for hepatic recurrence is unknown. METHODS: Fifty patients who underwent node dissection plus hepatectomy were retrospectively reviewed and divided into three groups: group I, 38 patients with a negative node; group II, 6 with a positive node at initial hepatectomy, and group III, 6 with a positive node at repeat hepatectomy. RESULTS: The 5-year survival rate after initial hepatectomy in group I was 46%. All patients in group II died within 2 years after surgery. In group III, the median survival time was 42 months after repeat hepatectomy, and 4 patients survived for more than 5 years after initial hepatectomy. Disease-free time was more than 1 year after initial hepatectomy in all long-term survivors. In addition, node metastasis was limited around the hepatic pedicle and postpancreatic area in 3 of 4 long-term survivors. CONCLUSIONS: Node dissection for lymphatic remetastasis may contribute to longer survival only when node metastasis is limited around the hepatic pedicle and postpancreatic area at repeat hepatectomy performed more than 1 year after the initial hepatectomy. PMID- 16721029 TI - Cloning, expression and regulation of chicken ovalbumin upstream promoter transcription factors (COUP-TFII and EAR-2) in the rat anterior pituitary gland. AB - Chicken ovalbumin upstream promoter transcription factors (COUP-TF)-II (NR2F2) and EAR-2 (NR2F6) are structurally related orphan members of the nuclear receptors superfamily. There are growing evidences that these factors play important roles during processes of differentiation and proliferation of several tissues. To better understand their role in the differentiated adult rat pituitary gland, we cloned COUP-TFII and EAR-2 cDNAs from an anterior pituitary cDNA library. Subsequently, we raised and characterized specific antibodies to the N-terminal domain of both nuclear receptors. We next examined their cellular and subcellular distribution in the pituitary gland and determined their regulation during pregnancy. COUP-TFII and EAR-2 pituitary genes display, respectively, 90 and 100% homologies with their human and mouse homologues. Cellular expression of both nuclear receptors was mainly detected in the lactotropes of male and female rats, with a prominent distribution in the nuclear compartment for EAR-2, and interestingly both proteins were significantly upregulated in pituitaries of pregnant vs. cycling female rats. Thus, our results have characterized cloning of rat pituitary COUP-TFII and EAR-2 genes, demonstrated that they are both specifically expressed in lactotropes, and strongly suggested that they may play an important role in modulating prolactin (PRL) gene expression during pregnancy. PMID- 16721031 TI - Differences in the appetite-stimulating effect of orexin, neuropeptide Y and ghrelin among young, adult and old rats. AB - Aging is associated with a progressive decrease in appetite and food intake. The appetite-stimulating peptides orexin A, neuropeptide Y (NPY) and ghrelin are known to play a critical role in food intake. In this study, the stimulatory effect of intracerebroventricular administration of these peptides on food intake was compared among young (4 months old), adult (11 months old) and old (24-27 months old) male Wistar rats. A stainless steel cannula was implanted stereotactically into the left lateral ventricle. After a 7-day recovery period, different doses of orexin A (0.25-3 nmol), NPY and ghrelin (0.03-1 nmol) were injected into the left lateral ventricle without anesthesia. Food consumption was measured at 1, 2 and 4 h after injection. We also examined the plasma levels of acylated and desacyl ghrelin in young and old rats by ELISA. Intracerebroventricular administration of orexin A and NPY stimulated food intake in young and adult rats, but no effects were observed at any dose in old rats. Ghrelin increased food intake in a dose-dependent manner in all groups, and the effect of ghrelin was reduced with advancing age. Neither the acylated nor the desacyl plasma ghrelin level differed significantly between young and old rats. In conclusion, the orexigenic effect of the peptides orexin A, NPY and ghrelin decreased in old rats, and this reduction may have been responsible for the age related decrease in food intake. PMID- 16721030 TI - Effects of ghrelin upon gonadotropin-releasing hormone and gonadotropin secretion in adult female rats: in vivo and in vitro studies. AB - A reproductive facet of ghrelin, a stomach-derived orexigenic peptide involved in energy homeostasis, has been recently suggested, and predominantly inhibitory effects of ghrelin upon luteinizing hormone (LH) secretion have been demonstrated in rat models. Yet, the modulatory actions of ghrelin on the gonadotropic axis remain scarcely evaluated. We report herein a detailed analysis of the effects of ghrelin upon LH and follicle-stimulating hormone (FSH) secretion in the female rat, using a combination of in vivo and in vitro approaches. Intracerebroventricular administration of ghrelin (3 nmol/rat) evoked a significant inhibition of LH secretion in cyclic female rats throughout the estrous cycle (proestrus afternoon, estrus, metestrus), as well as in ovariectomized females. In good agreement, gonadotropin-releasing hormone (GnRH) secretion by hypothalamic fragments from ovariectomized females was significantly inhibited by ghrelin. In contrast, ghrelin dose-dependently stimulated basal LH and FSH secretion by pituitary tissue in vitro; a phenomenon that was proven dependent on the phase of estrous cycle, as it was neither detected at estrus nor observed after ovariectomy. Conversely, GnRH-stimulated LH secretion in vitro was persistently inhibited by ghrelin regardless of the stage of the cycle, whereas stimulated FSH secretion was only inhibited by ghrelin at estrus. In addition, cyclic fluctuations in mRNA levels of growth hormone secretagogue receptor (GHS R)1a, i.e. the functional ghrelin receptor, were observed in the pituitary, with low values at estrus and metestrus. GHS-R1a mRNA levels, however, remained unchanged after ovariectomy. In summary, our data illustrate a complex mode of action of ghrelin upon the gonadotropic axis, with predominant inhibitory effects at central (hypothalamic) levels and upon GnRH-induced gonadotropin secretion, but direct stimulatory actions on basal LH and FSH secretion. Overall, our results further document the reproductive role of ghrelin, which might be relevant for the integrated control of energy balance and reproduction. PMID- 16721032 TI - Efficient human interferon-alpha gene transfer to neuroendocrine tumor cells with long-term and stable expression. AB - Interferon (IFN)-alpha has been used in the treatment of neuroendocrine (NE) tumors; however, the feasibility of IFN-alpha gene therapy has not been evaluated in NE tumor cells. In this study, human IFN-alpha2 (hIFN-alpha2) gene has been transferred into a NE tumor cell line BON. hIFN-alpha2-expressing BON cells were subcutaneously inoculated in nude mice. The results demonstrated that hIFN-alpha2 exerted significant antiproliferative effects on NE tumor cell lines (BON and LCC18) and other tumor cell lines (CA46 and SW480) as well as porcine aorta cell line. Furthermore, hIFN-alpha2 demonstrated its antineovascular activity in mice tumor and a direct antiangiogenic effect in chicken chorioallantoic membrane assay. hIFN-alpha2-expressing BON cells had a stable and long-term expression. Mice implanted with hIFN-alpha2-expressing BON cells showed a lower incidence, a delayed development and a significantly longer doubling time of the tumor compared to both wild-type (WT) and vector group. In addition, IFN-alpha significantly inhibited cell adhesion of WT BON cells. hIFN-alpha2-expressing BON tumors had a high level of hIFN-alpha2 protein. Finally, mice implanted with a mixture of WT and hIFN-alpha2-expressing BON cells (1:1) presented a delayed tumor development and had an even lower incidence of tumors than those implanted with hIFN-alpha2-expressing BON cells only. The doubling time of tumor was also longest in the mixture group. Our data suggest that hIFN-alpha2 gene therapy might be possible to be used as a new treatment for NE tumor patients. Further studies on the regulation of hIFN-alpha expression are needed, especially in combination with other cytokines, which could lead to a better understanding and improvements of hIFN-alpha gene therapy. PMID- 16721033 TI - Corticotropin-releasing hormone receptor expression on normal and tumorous human adrenocortical cells. AB - Corticotropin-releasing hormone (CRH) is not only the principal regulator of the central hypothalamic-pituitary-adrenal (HPA) axis but also exerts direct actions on peripheral tissues. We analyzed the expression of CRH receptors in microdissected preparations of normal human adrenal glands and in adrenocortical and adrenomedullary tumors, employing immunohistochemistry, quantitative RT-PCR of microdissected adrenal tissues, and in situ hybridization. The effect of CRH on adrenal steroidogenesis was tested in adrenal cells. Immunoreactive CRH1R was found primarily within the zona reticularis. In addition, we found a higher expression of CRH type-1 and 2 receptors mRNAs in preparations of adrenal cortices as compared to pheochromocytomas, a 6-fold increase in preparations of clinically unapparent adrenocortical adenomas, and a 10- to 60-fold increase in cortisol-producing adrenal adenomas. Stimulation of the adrenal tumor cell line NCI-H295R with CRH elicited a 1.4-fold increase in DHEA secretion. This result could be reproduced in a culture of primary human adrenocortical cells. We conclude that adrenocortical cells exhibit a higher expression of functional CRH receptors than chromaffin cells and that CRH acts on adrenal DHEA production. The data support the assertion of a direct action of CRH on human adrenocortical cells in addition to an intra-adrenal CRH receptor/adrenocorticotropin system. Enhanced CRH1R expression may be involved in adrenocortical tumorigenesis. PMID- 16721034 TI - Exercise enhances insulin and leptin signaling in the cerebral cortex and hypothalamus during dexamethasone-induced stress in diabetic rats. AB - Exercise and dexamethasone (DEX) are known to have opposite effects on peripheral insulin resistance. However, their effects and mechanism on brain glucose metabolism have been poorly defined. We investigated the modulation of the hypothalamo-pituitary-adrenal (HPA) axis and insulin/leptin signaling associated with glucose utilization in the brains of 90% pancreatectomized diabetic rats, which had been administered two dosages of DEX and exercised for 8 weeks. The data revealed that the administration of a high dose (0.1 mg/kg body weight/day) of DEX (HDEX) attenuated insulin signaling in the cerebral cortex and hypothalamus, whereas exercise potentiated their insulin signaling along with induction of IRS2 expression. In parallel with the modulated signaling, glucose utilization, such as glycogen storage and glycogen synthase activity, was suppressed by DEX in the cortex and hypothalamus, while exercise offset the DEX effects. Despite a decrease in epididymal fat mass, HDEX increased serum leptin levels, possibly due to an activated HPA axis, while exercise suppressed the increment. However, DEX reduced leptin-induced STAT3 phosphorylation in the cortex and hypothalamus, and it increased AMP-activated protein kinase (AMPK) phosphorylation only in the hypothalamus. Exercise reversed the phosphorylation of STAT3 and AMPK which had been modulated by DEX. In conclusion, exercise improves insulin and leptin signaling in the cerebral cortex and hypothalamus of diabetic rats exacerbated with HDEX, contributing to the regulation of body weight and glucose homeostasis. PMID- 16721035 TI - Analysis of the stress response in rats trained in the water-maze: differential expression of corticotropin-releasing hormone, CRH-R1, glucocorticoid receptors and brain-derived neurotrophic factor in limbic regions. AB - Glucocorticoids and corticotropin-releasing hormone (CRH) are key regulators of stress responses. Different types of stress activate the CRH system; in hypothalamus, CRH expression and release are increased by physical or psychological stressors while in amygdala, preferentially by psychological stress. Learning and memory processes are modulated by glucocorticoids and stress at different levels. To characterize the kind of stress provoked by a hippocampal dependent task such as spatial learning, we compared the expression profile of glucocorticoid receptor (GR), pro-CRH and CRH-R1 mRNAs (analyzed by RT-PCR), in amygdala, hippocampus and hypothalamus and quantified serum corticosterone levels by radioimmunoassay at different stages of training. mRNA levels of brain-derived neurotrophic factor (BDNF) were also quantified due to its prominent role in learning and memory processes. Male Wistar rats trained for 1, 3 or 5 days in the Morris water-maze (10 trials/day) were sacrificed 5-60 min the after last trial. A strong stress response occurred at day one in both yoked and trained animals (increased corticosterone and hypothalamic pro-CRH and CRH-R1 mRNA levels); changes gradually diminished as the test progressed. In amygdala, pro-CRH mRNA levels decreased while those of BDNF augmented when stress was highest, in yoked and trained animals. Hippocampi, of both yoked and trained groups, had decreased levels of GR mRNA on days 1 and 3, normalizing by day 5, while those of pro-CRH and CRH-R1 increased after the 3rd day. Increased gene expression, specifically due to spatial learning, occurred only for hippocampal BDNF since day 3. These results show that the Morris water-maze paradigm induces a strong stress response that is gradually attenuated. Inhibition of CRH expression in amygdala suggests that the stress inflicted is of physical but not of psychological nature and could lead to reduced fear or anxiety. PMID- 16721036 TI - Expression of mRNAs encoding receptors that mediate stress signals in gonadotropin-releasing hormone neurons of the mouse. AB - Neurons that synthesize and secrete gonadotropin-releasing hormone (GnRH) represent the neural control point for fertility modulation in vertebrates. As such GnRH neurons are ideally situated to integrate stress responses on reproduction. By isolating individual GnRH neurons from acute brain slices of adult female GnRH-EGFP transgenic mice and using microarray analyses, we have identified a range of transcripts encoding receptors known to be involved in stress responses in GnRH neurons. Prominent among these were receptors for corticotropin-releasing hormone (CRH), vasopressin, interleukins, prostaglandins, tumor necrosis factor alpha and other inflammatory mediators. We selected 4 of these targets [interleukin 1 receptor accessory protein (IL-1Racc), prostaglandin E(2) receptor subtype EP2 (PGER2), CRH receptor type 1 (CRH-R1), and arginine vasopressin receptor type 1b (AVP-R1b)] for validation using single-cell RT-PCR from individual GnRH neurons. In total, 54% of GnRH neurons (n = 26) were found to express at least 1 of these transcripts. The IL-1Racc, PGER2 and CRH-R1 mRNAs were each detected in approximately 25% of the GnRH neurons tested, but no evidence was found for AVP-R1b transcripts. Overlap was found between the expression of CRH-R1 and PGER2, and IL-1Racc and PGER2 in individual GnRH neurons. Dual immunofluorescence experiments confirmed the expression of CRH-R1/2 in a subpopulation ( approximately 30%) of GnRH neurons. These observations indicate that a variety of different stressors and stress pathways have the capacity to have an impact directly upon a subpopulation of GnRH neurons to influence the reproductive axis. PMID- 16721037 TI - The application of DIGE-based proteomics to renal physiology. AB - Proteomics is seeing increasing use as a means of identifying new mechanistic hypotheses in physiology. Proteomics based on two-dimensional electrophoresis (2 DE) has recently been optimized with the development of Difference Gel Electrophoresis (DIGE). In DIGE-based proteomics, the experimental and control samples are derivatized with different fluorophores and are run in the same gel, thereby minimizing technical variation. DIGE is currently one of the few techniques to perform quantitative proteomics, generating a statistical output to differences in protein abundances. In this review, we discuss the principles of DIGE-based proteomics, including sample preparation, 2-DE, statistical analysis of 2D-gels, and mass spectrometry. Strengths and weaknesses of DIGE are discussed, including possible solutions to overcome certain limitations, such as the identification of low abundance and integral membrane proteins. In addition, we provide a brief synopsis of our recent experiments in which DIGE-based proteomics was applied to study vasopressin signaling in the renal collecting duct. Finally, we illustrate how quantification based on the DIGE approach combined with bioinformatics may facilitate the study of systems biology of the kidney. PMID- 16721038 TI - Lobbying Congress to make cancer funding a priority: what scientists can do. PMID- 16721039 TI - A look back to move forward: scientists in reflections. PMID- 16721040 TI - BRCA1 phosphorylation: biological consequences. AB - More than a decade has passed since BRCA1, breast cancer tumor suppressor 1, was isolated by reverse genetics in 1994. Its molecular structure and potential function have been extensively studied; both mouse genetics and a cell culture system revealed that BRCA1 is a 220,240 kD nuclear phosphoprotein, it regulates transcription, its loss leads to genome instability and in turn, cell transformation. Significantly, DNA checkpoint associated kinases have been shown to phosphorylate specific residues of BRCA1 under conditions of DNA damage, making cells sensitive or resistant to various stresses. Our recent findings support the notion that UV induced phosphorylation of particular residues of the protein is crucial for activation of caspase 3. This article will focus on the BRCA1 kinases, the identification of the phosphorylation residues, and the biological consequences of BRCA1's phosphorylation for regulation of cell proliferation. PMID- 16721041 TI - Questioning the oncogenic role of DeltaNp73alpha in different cell lines expressing p53 or not. AB - The recent finding that the 1p36.3 locus gene encodes an array of different p73 isoforms with apparently distinct and sometimes opposing cellular functions, might explain the difficulty in establishing the protein's role as tumor suppressor. Therefore we need to investigate the roles of each of these splicing variants in cellular functions when expressed alone or in combination with other family members, as well as the genetic background on which the proteins are expressed. We investigated, in two p53 null cell lines, the human SCLC line H1299 and a subline derived from the human colon carcinoma cell line HCT116 (HCT116/379.2), the effects of DeltaNp73alpha overexpression on cell growth and the response to anticancer treatment. We generated three different clones overexpressing DeltaNp73alpha under a tetracycline inducible promoter. Immunofluorescent staining and luciferase reporter assays confirmed that clones HCT116/DeltaNA and H1299/DeltaN7 and H1299/DeltaN11 did express a functional, nuclear localized DeltaNp73alpha protein. The stable overexpression of DeltaNp73alpha protein did not confer any cell growth advantage. Doubling time of clones overexpressing DeltaNp73alpha were comparable to counterparts not expressing it. Clonogenic assays showed that the cytotoxic activity of different DNA damaging agents, such as cDDP, UV light and doxorubicin, were comparable in clones expressing DeltaNp73 or not. The overall data argue against an oncogenic role for this isoform. These findings are independent of the p53 status since they overlap with those previously obtained by our group in HCT116 cell lines, wild type for p53. PMID- 16721042 TI - NF-kappaB does not influence the induction of apoptosis by Ukrain. AB - Ukrain is a reaction product of different alkaloids from Chelidonium majus L. (celandine) conjugated with thiophosphoric acid. It has immunoregulatory effects on T lymphocyte subsets and cytotoxic and cytostatic effects on various malignant cells. Although Ukrain has been reported to induce alterations in the cell cycle and tubulin polymerization, the specific cellular target has not been described. Since antineoplasic agents induce NF-kappaB and their effects are regulated by this transcription factor, we investigated its possible participation in the apoptotic effects of Ukrain. Ukrain induced apoptosis in a panel of cancer cell lines by activating the intrinsic cell death pathway, as demonstrated by the cleavage of caspase 9 and the upregulation and cleavage of caspase 3. The effect was reversible, since long exposures (24 hours or more) were needed, as verified by clonogenic assays. Gene reporter assays showed that Ukrain activated NF-kappa B. Nevertheless, this activation was not required for, and did not modulate, the Ukrain effect: neither blockage of activation by a dominant negative version of Ikappa-B alpha or a Bcl-3 siRNA, nor activation of the pathway by overexpression of IKK2, changed the response to the drug. In conclusion, Ukrain induced apoptosis in HeLa cervical cancer cells by activating the intrinsic pathway. In contrast to other antineoplasic drugs, the effects of Ukrain were not regulated by NF-kappa B. PMID- 16721043 TI - Sequence mutations and amplification of PIK3CA and AKT2 genes in purified ovarian serous neoplasms. AB - Sequence mutations and gene amplifications lead to activation of the PIK3CA-AKT2 signaling pathway and have been reported in several types of neoplasms including ovarian cancer. Analysis of such genetic alterations, however, is usually complicated by contamination of normal cell DNA, artifacts associated with formalin-fixed tissues and the sensitivity of the techniques employed. In this study, we analyzed the sequence mutations in PIK3CA and AKT2 genes using purified tumor cells that were isolated from high-grade ovarian serous carcinomas and serous borderline tumors (SBTs) and assessed gene amplification using a dual color FISH on tissue microarrays. Somatic sequence mutations in the kinase domain of AKT2 were not detected in any of the 65 ovarian tumors analyzed. Mutations of PIK3CA were rare, occurring only in one (2.3%) of 44 high-grade serous carcinomas and in only one (4.8%) of 21 SBTs. Dual-color FISH demonstrated that PIK3CA and AKT2 were not amplified in SBTs but amplified in 13.3% and 18.2% high-grade carcinomas, respectively. High-level amplification (>3 fold) was more frequently observed in AKT2 than in PIK3CA. Unlike mutations in ERBB2, KRAS and BRAF which are mutually exclusive in SBTs, coamplification of PIK3CA and AKT2 was present in five high-grade carcinomas including the OVCAR3 cells. Amplification in either of the genes occurred in 27% high-grade serous carcinomas. In conclusion, the methods we employed provide unambiguous evidence that somatic sequence mutations of PIK3CA and ATK2 are rare in ovarian serous tumors but amplification of both genes may play an important role in the development of high-grade ovarian serous carcinoma. PMID- 16721044 TI - Heterogeneous cross-talk of E2F family members is crucially involved in growth modulatory effects of interferon-gamma and EGF. AB - There is growing evidence that deregulation of E2F transcription factors is causatively involved in the patho-physiology of various tumors. However, no data on the role of E2F family members in tumor biology of ovarian cancer are available. We here describe an expression study of all known E2F transcription factors and their coactivators DP-1 and DP-2 in various human ovarian cancer cell lines and the breast cancer cell line T47D and their involvement in pathways affected by interferon-gamma and EGF. A significant overexpression of the proliferation-promoting E2F1 and especially E2F2 points to a pivotal role in modulating the uncontrolled proliferation in ovarian cancer cells. Of special note is the fact that interferon-gamma treatment did not only caused a reduction of the proliferation-promoting transcription factors E2F1 and E2F2, but also increased the inhibiting transcription factors E2F4 and E2F5, thus underlining the importance of an E2F cross-talk in the anti-proliferative function of interferon-gamma. Moreover, an increase in DP-1 and E2F3 is probably involved in the proliferation-enhancing effect of EGF. Our study provides a new insight in the crucial role of E2F cross-talk, especially the role of the inhibiting transcription factors E2F4 and E2F5, in the tumor biology of cancer and its possible usefulness as targets in anti-cancer therapy. PMID- 16721045 TI - Kaposi's sarcoma-associated herpesvirus LANA hitches a ride on the chromosome. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) latently infects tumor cells and has an etiologic role in Kaposi's sarcoma, primary effusion lymphoma, and multicentric Castleman's disease. Survival in rapidly dividing cells depends on a carefully orchestrated chain of events. The viral genome, or episome, must replicate in concert with cellular genetic material, and then efficiently segregate to progeny nuclei. KSHV achieves this through its latency associated nuclear antigen (LANA), which simultaneously binds to viral DNA and mitotic chromosomes to efficiently partition episomes. LANA's N-terminal region has been shown to be essential for efficient KSHV DNA replication and tethering to mitotic chromosomes. The precise mechanism by which LANA attaches to host chromosomes has been an area of active investigation. We recently reported that this association is mediated by the chromatin components histones H2A and H2B. Binding between LANA and these proteins was demonstrated in vivo and in vitro, and use of an H2A H2B depleted system demonstrated their central role in LANA's chromosome binding. Further, we provided a structural description of the interaction of LANA's N terminal chromosome association region with the nucleosome using x-ray crystallography. Our data offer further insight into the mechanism of KSHV latency, and also reveal a new concept for a role of the nucleosome as a docking site for other proteins. PMID- 16721046 TI - The loss of gammaH2AX signal is a marker of DNA double strand breaks repair only at low levels of DNA damage. AB - The induction of DNA double-strand breaks (DSBs) by genotoxic treatment leads to high toxicity and genetic instability. Various approaches have been undertaken to quantify the number of breaks and to follow the kinetic of DSB repair. Recently, the phosphorylation of the variant histone H2AX (named gammaH2AX), quantified by specific immunodetection approaches, has provided a valuable and highly sensitive method to monitor DSBs formation. Although it is admitted that the number of gammaH2AX foci reflected that of DSBs, contradictory reports leave open the question of a link between the disappearance of gammaH2AX signal and DSBs repair. We determined gammaH2AX expression (i) in cells either proficient or not in DSBs repair capacity, (ii) after exposure to ionizing radiation (IR) or calicheamicin gamma1, a radiomimetic compound, (iii) and by three different immunodetection methods, foci numbering, flow cytometry or Western blotting. We showed here that gammaH2AX loss correlates with DSB repair activity only at low cytotoxic doses, when less than 100-150 DSBs breaks per genome are produced, independently of the method used. In addition, in DNA repair proficient cells, the early decrease in the number and intensity of gammaH2AX foci observed after a 2 Gy exposure was not associated with a significant change in the global gammaH2AX level as determined by Western blotting or flow cytometry. These results suggest that the dephosphorylation step of gammaH2AX may be limiting and that the loss of foci is mediated not only by gammaH2AX dephosphorylation but also through its redistribution towards the chromatin. PMID- 16721047 TI - Repression of new p53 targets revealed by ChIP on chip experiments. AB - Following DNA-damage, the tumor suppressor p53 activates G1/S blocking and apoptotic genes, and represses other genes, including those involved in G2/M transition. In this latter system, it acts through the CCAAT-binding histone-like NF-Y. Several groups have reported that p53 is associated to promoters in unstressed conditions. We developed an oligo-based array containing 179 human promoters, enriched in genes involved in the DNA-damage and ER-stress response. We performed ChIP on chip experiments with p53 and NF-Y in cells under normal growing conditions. We identified 46 new p53 targets and noted (i) a significant enrichment in genes of the ER-stress response, including crucial regulators such as XBP1 and C/EBPbeta (ii) genes whose products are involved in the regulation of p53 function. Several genes were validated by conventional ChIP. DNA-damage dependent PCAF-mediated acetylation was observed on most, but not all promoters. The effect of p53 activation was checked by RT-PCR and transfections in HCT116 wt, E6 and p53-/- cells: most promoters were actively repressed upon Adriamycin treatment or following p53 transfection in p53-/- cells. In particular, the behaviour of some of the genes (BRCA1, RAD23 and RAD17) is consistent with a feedback loop regulation on p53 levels. Finally, there is a large overlap (66%) between p53 and NF-Y targets. Our data reinstate the physiological importance of p53 promoter recognition and direct transcriptional repression as a mechanism to cope with DNA-damage. PMID- 16721048 TI - Theoretical proposal: allele dosage of MAP2K4/MKK4 could rationalize frequent 17p loss in diverse human cancers. AB - Although aneuploidy is a global genomic abnormality present in most human cancers, the clonal selection model best explains the action of select activating mutations in oncogenes and homozygous losses of tumor-suppressor genes. Simple gene dosage changes are difficult however, to incorporate into this model, in part due to negative feedback loops that govern major cancer mutational targets (e.g., TP53, PTCH, SMAD4) and essentially preclude a haploinsufficient phenotype. The 17p conundrum may offer a clue to reconciling this difficulty: In comparison to the moderate mutation rate of TP53, many tumors have a disproportionately high frequency of loss of 17p. This discrepancy, and similar discrepancies at other sites of LOH, has long been thought to be due to the presence of undiscovered yet frequently mutated tumor-suppressor genes. However, over 15 years of searching for this grail has distributed bountiful disappointment. It is perhaps time to seriously consider an alternative explanation. Located on 17p adjacent to the TP53 gene, MKK4 is one of the most consistently mutated genes across tumor types, and is located on one of the most frequently lost arms in the human genome. We theorized that a gene dosage-dependent phenotype of MKK4 could plausibly promote the emergence of 17p LOH and thereby the probability of evolving the biallelic inactivation of TP53. Using MKK4 somatic human knockout cancer cells, we observed the proof-of-principle in the downstream gene dosage-dependent phenotypes: heterozygous and homozygous knockouts were progressively deficient in Mkk4 protein, in stress-induced phosphorylation of Jnk, and the resultant upregulation of JUN mRNA. These observations highlight a lack of compensatory regulation when gene dosage changes perturb the Jnk-Jun relationship. Consideration of gene dosage changes specifically affecting members of positive feedback loops may permit integration of the aneuploidy process into a conventional model of clonal selection in tumorigenesis. PMID- 16721049 TI - Double bolt regulation of Rad51 by p53: a role for transcriptional repression. AB - The regulation of homologous recombination (HR) by p53 has been currently associated with its nontransactivating function; now the transcriptional repression of the Rad51 gene by p53 has been elucidated. This extra control by p53 involves the transcriptional downregulation of Rad51 protein, which in turn accounts for the reduction of functional Rad51 foci. Down regulation of Rad51 protein might be relevant, considering that Rad51 is a rate-limiting factor in HR. Moreover, impaired Rad51-repression by p53 mutant proteins may contribute to malignant transformation. PMID- 16721050 TI - GDF3 at the crossroads of TGF-beta signaling. AB - Embryonic stem (ES) cells present an excellent system for addressing the relevance of our current knowledge about how cell fate is determined and how cells integrate multiple signals into a single outcome as a function of time. Many of the factors that mediate these phenomena have been discovered through classical embryological experiments and are organized into several major signal transduction pathways including TGF-beta/BMP, Jak-STAT, Hedgehog, Wnt, Notch and FGF/MAPK. This review will summarize the current understanding of TGF-beta signaling in ES and focus on early embryological roles of the TGF-beta member, GDF-3. GDF-3 is associated with the undifferentiated state of ES cells and two recent and contradictory reports examined the function and mechanism of GDF-3 in the context of both stem cells and early embryonic differentiations. While Levine and Brivanlou found that GDF-3 inhibits its own subfamily members (the BMPs), Chen and colleagues found that GDF-3 acts as a nodal-like TGF-beta ligand. These combined findings raise the intriguing possibility that GDF-3 acts as a bi functional protein, to regulate the balance between the two modes of TGF-beta signaling. PMID- 16721051 TI - UV-mediated NF-kappaB activation is abolished in deficient XPC/D primary fibroblasts. AB - Genetic pathologies involving deficits in DNA repair, like xeroderma pigmentosum (XP), show recurrent cell death, tissue degeneration and oncogenesis due to high sensitivity to ultraviolet radiation (UV). Various inducers including UV activate NF-kappaB, a pathway largely involved in cell proliferation and apoptosis. However, the mechanism(s) involving NF-kappaB activation by UV are poorly understood. To improve this knowledge, we examined NF-kappaB in two XP cell groups (XPC and XPD/TTD). XPC/D primary fibroblasts possess functional NF-kappaB dimers, and pro-inflammatory cytokines consistently activate NF-kappaB pathway. Contrarily, UV-mediated NF-kappaB activation is practically absent, whereas kappaB-specific DNA binding and transcriptional activity are dramatically undermined. These results indicate that lack of UV responsiveness at the NF kappaB level is a common feature of XPC/D cells, suggesting that XP proteins might act upstream on NF-kappaB activity induced by UV. These observations help us to better understand the UV sensitivity and compromised survival of XP deficient cells. PMID- 16721052 TI - The TRIP-Br family of transcriptional regulators is essential for the execution of cyclin E-mediated cell cycle progression. AB - The G1 D-type cyclins, in conjunction with cyclin-dependent kinases Cdk4 and Cdk6, play key roles in the execution of mitogen-induced cellular proliferation. TRIP-Br1, a member of the TRIP-Br family of transcriptional regulators, has been implicated in the regulation of Cdk4/cyclin D activity. To further elucidate the functional role(s) of the TRIP-Br proteins in mitogenic signaling, we have developed the synthetic DNA enzymes E-Br1 and E-Br2 to specifically knock down the serum-inducible expression of TRIP-Br1 and TRIP-Br2, respectively, in WI-38 human fibroblasts in culture, as well as generated TRIP-Br2 null primary embryonic fibroblasts from a novel TRIP-Br2 knockout mouse model. Both strategies consistently reveal that ablation of TRIP-Br1 or TRIP-Br2 expression disrupts mitogenic signaling in a manner that suppresses serum-induced cyclin E expression, S-phase entry and cellular proliferation. We conclude that both TRIP Br1 and TRIP-Br2 are required for proper transduction of mitogenic signals and execution of serum-inducible cell cycle progression. PMID- 16721053 TI - Spreading the signal: dissociation of Chk1 from chromatin. AB - The effector kinase Chk1 plays a critical role in the DNA damage checkpoint response by phosphorylating regulators of the cell cycle machinery, resulting in an inhibition of cell cycle progression. In the presence of genotoxic stress, the PI3 kinase-like kinase ATR rapidly phosphorylates Chk1 on conserved serine residues, thereby triggering kinase activation through the release of an auto inhibitory region present at its C-terminus and by regulating interactions with other proteins. Recent data have demonstrated an additional regulatory mechanism of Chk1 functioning: Chk1 binds chromatin in unperturbed cells and dissociates from chromatin in response to DNA damage in a manner that is dependent on PIKK mediated phosphorylation. Here, I give an overview of these findings and discuss the implications of these data for our understanding of the execution of the checkpoint arrest induced after the detection of DNA damage. PMID- 16721055 TI - From description to causality: mechanisms of gene expression signatures in cancer. AB - Global gene expression profiles of thousands of cancer samples have been completed, giving rise to hundreds of gene expression signatures (GES). Although many expression signatures show promise in predicting patient prognosis or response to therapies, the usefulness of the signatures in understanding the underlying mechanisms of cancer has not been fully exploited. While "reverse genomic" methods can test specific hypotheses of gene regulation, they fare less well in deciphering novel or combinatorial mechanisms of gene regulation. Recently we described SLAMS (stepwise linkage analysis of microarray signatures), a novel method that can prospectively identify genetic regulators of gene expression signatures in cancer. Applying SLAMS on a poor-prognosis wound signature in human breast cancer, we identified CSN5-mediated ubiquitination of MYC as a novel mechanism to activate a biological program favoring metastasis. PMID- 16721054 TI - Bur1/Bur2 and the Ctk complex in yeast: the split personality of mammalian P TEFb. AB - Eukaryotic organisms possess a host of factors that regulate transcriptional elongation. In higher eukaryotes, the transcription factor P-TEFb not only regulates phosphorylation of the RNA polymerase II C-terminal domain, but it also inhibits the action of transcriptional repressors and is required for the association of several elongation factors with the transcribing polymerase. In the yeast Saccharomyces cerevisiae, the cyclin dependent kinases Bur1/Bur2 and Ctk complex (Ctk1, 2 and 3) are also able to impact several aspects of transcription. Together, these two kinase complexes appear to functionally reconstitute the activity of P-TEFb in yeast. Recent findings regarding the role of these kinases in histone tail modifications and transcriptional regulation is briefly reviewed below. PMID- 16721056 TI - G1/S Cyclins interact with regulatory subunit of PKA via A-kinase anchoring protein, AKAP95. AB - The mechanisms by which cyclins promote mammalian cell cycle progression have been a topic of intense investigation over the last decade. We previously described an interaction between D-type cyclins and A-kinase anchoring protein, AKAP95. Here, we demonstrate that AKAP95 can also bind cyclin E1. Association between AKAP95 and cyclins is displaced by CDKs. We show that these G(1)/S cyclins can interact with RII subunit of PKAalpha through AKAP95. The presence of alternate complexes cyclin-CDK and cyclin D/E-AKAP95-PKA.RIIalpha suggest different roles of G(1)/S cyclins and a wider biological importance of these interactions in cells. PMID- 16721057 TI - DEK Expression is controlled by E2F and deregulated in diverse tumor types. AB - Deregulation of the retinoblastoma (pRB) tumor suppressor pathway associated with aberrant activity of E2F transcription factors is frequently observed in human cancer. Microarray based analyses have revealed a large number of potential downstream mediators of the tumor suppressing activity of pRB, including DEK, a fusion partner of CAN found in a subset of acute myeloid leukaemia (AML) patients carrying a (6; 9) translocation. Here we report that the expression of DEK is under direct control of E2F transcription factors. Chromatin immunoprecipitation assays show that the DEK promoter is bound by endogenous E2F in vivo. The DEK promoter is transactivated by E2F and mutation of E2F binding sites eliminates this effect. Expression levels of DEK in human tumors have been investigated by tissue micro array analysis. We find that DEK is overexpressed in many solid tumors such as colon cancer, larynx cancer, bladder cancer, and melanoma. PMID- 16721058 TI - Multiple initiation sites within the human ribosomal RNA gene. AB - Numerous studies have demonstrated that DNA replication initiates within the 30 kB non-transcribed spacer (NTS) region of the human ribosomal RNA gene (rDNA). Using a series of closely spaced primer pairs to measure nascent leading strand abundance in mid and late S phase cells isolated by centrifugal elutriation, we find evidence for one highly preferred initiation site and two less utilized sites within a 6 kb region of the NTS. The initiation sites colocalize with significant DNA unwinding elements (DUEs), matrix attachment regions (MARs), and ARS-like sequences. An intrinsic DNA bending site was localized by circular permutation analysis to within several hundred base pairs of one initiation site. While DUE and MAR elements occur elsewhere throughout the 43 kb rDNA sequence, the close association of DUE and MAR elements occurs only near replication initiation sites, a juxtaposition also seen in other well-studied mammalian replication initiation sites. The utilization of rDNA initiation sites close to DUE and MAR elements in mid and late S phase, but not in very early S phase as previously shown, suggests that in rRNA genes, contributions from these sequence associated properties may be more significant to initiation sites associated with transcriptionally inactive genes, than to initiation sites associated with transcriptionally active genes. PMID- 16721059 TI - Live free or die: an immature T cell decision encoded in distinct Bcl-2 sensitive and insensitive Ca2+ signals. AB - In the developing thymus, strong T cell receptor (TCR) activation by self antigens induces negative selection and weak TCR activation induces positive selection. Both processes are mediated by Ca(2+) signals, raising the question of how a single second messenger like Ca(2+) can mediate such diverse cell fates. Recent findings indicate that graded TCR activation signals are encoded in distinct patterns of Ca(2+) elevation. The anti-apoptotic protein Bcl-2 discriminates between these Ca(2+) signaling patterns, selectively inhibiting pro apoptotic Ca(2+) signals induced by strong TCR activation without suppressing pro survival Ca(2+) signals induced by weak TCR activation. PMID- 16721060 TI - Y-box binding protein 1: providing a new angle on translational regulation. AB - Current models of translational regulation are mostly focused on how translational factors engage a messenger mRNA to the ribosome to initiate translation. Since the majority of mRNAs in eukaryotes are translated in a cap dependent manner, the mRNA 5' cap-binding protein eIF4E was characterized as a key player responsible for the recruitment of mRNAs to the initiation complex. The availability of eIF4E is believed to be especially critical for translational activation of mRNAs with extensive secondary structures in their 5'UTRs, many of which code for labile regulatory proteins essential for cell growth or viability. Surprisingly, little attention is paid to the other side of translational control, e.g., to define mechanisms responsible for translational silencing and storage of the above messages. In this review, we discuss the possibility that eIF4E per se may not be sufficient to release mRNAs from translational block. We found that many growth- and stress-related mRNAs are associated with the translational repressor YB-1, which can compete with the eIF4E-driven translation initiation complex for binding to the capped 5' mRNA terminus. Moreover, the cap dependent repressor activity of YB-1 appears to be negatively regulated via Akt mediated phosphorylation of the Ser-102 residue of YB-1. Taken together with recent evidence suggesting that translational activation of growth-related messages is a primary cellular response to activation of Ras-Erk and PI3K-Akt signaling pathways, our data suggest that differential expression of specific mRNA subsets is regulated by the PI3K-Akt pathway and achieved via coordinated activation of the components of translational machinery and inactivation of general translational repressors such as YB-1. PMID- 16721061 TI - Exploring the positive and negative consequences of NF-kappaB inhibition for the treatment of human disease. AB - The NF-kappaB (NF-kappaB) transcription factor-signaling pathway has become a major focus for research aimed at understanding its role in development, maintenance and progression of disease. A large body of recent research suggests inhibition of NF-kappaB blocks inflammation, cancer development and progression, diabetes, stroke, muscle wasting, and other diseases. The enormous potential for the treatment of disease by inhibiting NF-kappaB lead to the development of inhibitory drugs that specifically target this pathway. At the same time offering great clinical potential, inhibition of NF-kappaB in vivo can be detrimental. NF kappaB controls multiple functions in homeostasis including a functional immune response, cell cycle, and cell death. Genetic studies in mice and analysis of naturally occurring mutations in humans point to specific developmental and immune consequences due to altering NF-kappaB activity. The balance between therapeutic benefit and potential changes in normal cellular function and response during drug induced NF-kappaB inhibition will be one of the challenges in future clinical studies. PMID- 16721063 TI - The role of polycomb group proteins in cell cycle regulation during development. AB - Polycomb group (PcG) and trithorax group (trxG) proteins are evolutionarily conserved chromatin modifiers that have well known roles in the maintenance of silent and active expression states of homeotic genes. PcG proteins may also be involved in the control of cellular proliferation, as several PcG complexes have been shown to act either as proto-oncogenes or as tumor suppressors in vertebrates. In Drosophila, PcG factors associate with specific DNA regions termed PcG response elements (PREs), and a PRE was recently identified in the gene encoding Cyclin A. Still, it is not yet clear how PcG complexes could control cell cycle progression. Beyond acting as stable silencers of cell cycle genes during the differentiation process, PcG complexes might also be integrators and/or modulators of cell cycle checkpoints in dividing cells. Here, we discuss this dual aspect of PcG involvement in epigenetic cell cycle control. PMID- 16721062 TI - Mechanistic insights into the regulation of the spermatogonial stem cell niche. AB - Potential therapeutic use of stem cells in the treatment of human diseases depends on our ability to control the balance of their differentiation and self renewal in vitro and in vivo. The stem cell "niche," or specialized microenvironment, is now recognized as one of the major contributors to this regulation in many species. Our recent study, which was reported in Nature, was the first to demonstrate that expression of a vertebrate animal transcription factor is essential for the maintenance of a stem cell niche. In that letter, targeted disruption of ERM (Ets-related molecule), which was localized only in the somatic support cell of the testis, the Sertoli cell, resulted in failure of self-renewal by spermatogonial stem cells, following the first wave of spermatogenesis. One of the more important conclusions drawn was the realization that regulation of the stem cell niche during the perinatal period, a phase characterized by rapid mitosis of both spermatogonial stem cells and Sertoli cells, differed from that in the adult. It appears that the ERM-regulated pathways are coincident with the termination of Sertoli cell proliferation and commencement of the cycle of spermatogenesis, which is sustained by the same cell that regulates the stem cell niche. Several likely targets for ERM regulation are discussed, as well as their potential implications for increasing our understanding of spermatogonial stem cell activity and the uniqueness of the Sertoli cell's immune privilege and possible utility for the protection of transplanted adult stem cells. PMID- 16721065 TI - Minichromosome maintenance proteins as markers for proliferation zones during embryogenesis. AB - Regulation of cell proliferation is of fundamental importance for growth and prevention of cancer. An obligatory step in the regulation of cell proliferation is the control of the initiation of DNA synthesis. The minichromosome maintenance (MCM) proteins are essential DNA replication factors crucial for initiating DNA synthesis once every cell cycle. Recent studies show that the level of MCM proteins is stringently regulated to correlate with cell proliferation and carcinogenesis. Here we discuss recent data, which highlights the usefulness of minichromosome maintenance (MCM) gene expression for detecting proliferating cells as well as zones containing proliferative/stem cells during embryogenesis in a whole organism. PMID- 16721064 TI - Lipid modification of secreted signaling proteins. AB - Proteins of the Hedgehog, Wnt and Epidermal Growth Factor Receptor (EGFR) ligand families are secreted signals that induce concentration-dependent responses in surrounding cells. Although these proteins must diffuse through the aqueous extracellular environment, recent work has shown that hydrophobic lipid modifications are essential for their functions. All three classes of ligands are palmitoylated in the secretory pathway by related enzymes, and Hedgehog also carries a C-terminal cholesterol modification as a result of its autocatalytic cleavage. Palmitoylation is required for Wingless secretion and contributes to the signaling activity of Hedgehog and Wnt3a, but is not required for secretion or receptor activation by the EGFR ligand Spitz. While lipid modifications enhance the long-range activity of Sonic hedgehog, they restrict the range and increase the local concentration of Spitz. We discuss the diverse functions and the possible extent of palmitoylation of secreted ligands. PMID- 16721066 TI - A novel guanine nucleotide exchange factor MyoGEF is required for cytokinesis. AB - The cleavage furrow is created by an actomyosin contractile ring that is regulated by small GTPase proteins such as Rac1 and RhoA. Guanine nucleotide exchange factors (GEFs) are positive regulators of the small GTPase proteins and have been implicated as important factors in regulating cytokinesis. However, it is still unclear how GEFs regulate the contractile ring during cytokinesis in mammalian cells. Here we report that a novel GEF, which is termed MyoGEF (myosin interacting GEF), interacts with non-muscle myosin II and exhibits activity toward RhoA. MyoGEF and non-muscle myosin II colocalize to the cleavage furrow in early anaphase cells. Disruption of MyoGEF expression in U2OS cells by RNA interference (RNAi) results in the formation of multinucleated cells. These results suggest that MyoGEF, RhoA, and non-muscle myosin II act as a functional unit at the cleavage furrow to advance furrow ingression during cytokinesis. PMID- 16721067 TI - A comparison of ipsilateral pedicled TRAM flap with and without previous irradiation. AB - Due to concerns for increased complications, there is reluctance to use the ipsilateral pedicled TRAM for breast reconstruction in the presence of previous chest wall irradiation. This study will assess whether the ipsilateral pedicled TRAM is a safe and effective option when the pedicle is part of the irradiation field. Consecutive cases of ipsilateral pedicled TRAM flap procedures from 1997 2003 were reviewed. Patients with and without previous irradiation were compared on several vascular indicators. There were 123 and 124 patients in the irradiated and nonirradiated groups, respectively. These groups were demographically similar, except the nonirradiated group was significantly older. The irradiated group had a significantly higher rate of minor wound problems related to mastectomy flap healing (8.9% versus 1.6%). All other flap vascular complications were equivalent. The ipsilateral pedicled TRAM flap is a safe and effective option for breast reconstruction in an irradiated field in terms of the pedicle to the flap. PMID- 16721069 TI - An alternative fixation technique for the endoscopic brow lift. AB - BACKGROUND: Endoscopic brow lift has become widely accepted as a procedure for restoring a youthful brow, since only 3 hardly noticeable incisions of the scalp are needed for this subperiosteal dissection and final repositioning of the brow. It has become an acceptable technique, an alternative to the conventional technique or transcoronal browpexy. One of the controversial points is the fixation of the flap in the elevated position. METHOD: Endoscopic brow lift allows separation and repositioning of the periosteum of the orbital rims and zygomaxilla. In a 6-year period from September 1999, 300 patients underwent endoscopic brow lift using our fixation approach, which was accomplished with an absorbable suture subperiosteally. RESULTS: Satisfactory forehead rejuvenation was obtained in all patients, with correct eyebrow movement. CONCLUSIONS: Long term results of 6 years confirm the strength and durability of this fixation approach. We consider this approach to be a simple, secure, and reliable forehead fixation method, an alternative to other fixation methods, that allows satisfactory and long-lasting cosmetic results. PMID- 16721068 TI - Institutional review of free TRAM flap breast reconstruction. AB - INTRODUCTION: A 10-year experience with breast reconstruction in a university hospital was recently reviewed. The purpose of this study was to determine the subtypes of breast reconstructive procedures and to evaluate the frequency and change in technique over time of free TRAM (transverse rectus abdominis muscle) flap breast reconstruction performed at one institution. Trends in the development of the procedure over this period were also reviewed. MATERIALS & METHODS: Between November 1994 and September 2004, a 10-year retrospective chart review was conducted. The indications for mastectomy and reconstruction were determined. The median age was 48 (range 31-66). The range of follow-up was 2 to 71 months, with a median of 19.5 months. The mean follow-up was 23.5 months. Outcome data were grouped into 2 consecutive 5-year periods (period 1: 1994-1998; period 2: 1999-2004) and evaluated for changes over time in techniques and outcome. Statistical analysis (Decision Analyst, Inc., STATS Statistics software, version 1.1, 1998) was performed using the difference between 2 proportions module to assess the probability of a significant difference in the data for period 1 and period 2 parameters. RESULTS: Over a 10-year period, 117 patients underwent breast reconstruction. This consisted of 12 pedicle procedures (11.3%), including 1 bipedicle flap (0.9%) and 2 bilateral pedicle procedures (1.8%). There were 3 latissimus dorsi pedicle flaps (2.8%). Sixteen patients (15.1%) received tissue expander or implant reconstructions. Of the 117 patients, 79 underwent free flap breast reconstruction. Of the 79 free-flap patients, 22 (27.8%) had bilateral procedures, for a total of 101 free flaps performed in these 79 patients. Fifty-two patients underwent immediate reconstruction (65.8%) and 25 were delayed (31.6%) reconstructions using either deep inferior epigastric artery perforator (DIEP) flaps (4 = 3.9%) or free TRAM flaps (97 = 96.0%). A muscle-sparing technique was used in 43 of the 97 free TRAM flaps (44.3%). The preferred vascular inflow was the internal mammary artery, which was used in 66 out of 101 flaps (65.3%). The rate of anastomotic revision (arterial and venous) was 4.9%. The majority of cases used a 2.5-mm venous coupler (65.3%). In 2 of the free TRAM cases, there was insufficient volume to establish the patients preexisting volume. Therefore, at the patient's request, immediate implants were used to augment the reconstruction. The average hospital stay was 8.13 days, and the average intensive care stay was 4.59 days. When assessed for trends over time, we noted a reduction in our hospital length of stay and our ICU length of stay. CONCLUSION: The experience with free tissue breast reconstruction reveals predominant use of the TRAM flap. This is justified by the reliability of this flap and the advances in achieving esthetic breast reconstruction. Additionally, we have begun performing DIEP free-flap reconstructions. Our clinical practice has evolved concurrent with standards of care, as noted by the increase in use of muscle-sparing techniques and the reduction in the use of dextran. We do not routinely use therapeutic anticoagulation in our cases. Our hospital length of stay and average intensive care length of stay have also decreased over time, consistent with a system-wide effort to increase the efficiency of healthcare delivery. PMID- 16721070 TI - V-Y in V-Y procedure: new technique for augmentation and protrusion of the upper lip. AB - A new surgical technique for upper lip augmentation is presented. In this technique, 2 vertically parallel V-shaped incisions placed in a "V in V" fashion are used. Remarkable augmentation of the lip is provided by V-to-Y closure of the incisions. Over 7 years, this new technique, namely, V-Y in V-Y procedure, was used in 46 female patients for upper lip augmentation. The mean age of patients was 23 years, ranging from 16 to 47 years. The average follow-up was 2 years. In all patients, a satisfactory upper lip augmentation was obtained. Except a moderate edema subsiding within 2-3 weeks postoperatively, there was no complication in our patients. Despite that no filling material was used, the results were found to be long lasting at late term. The "V-Y in V-Y" procedure is a very easygoing and adjustable technique which allows the surgeon to augment the lip tubercle and the lateral segments separately, as required by the individual needs of each patient. Besides an anatomic lip augmentation by preserving and accentuating the natural contours of the lip, this new procedure provides a remarkable protrusion of the entire upper lip as an additional contribution for rejuvenation of the upper lip. PMID- 16721071 TI - Reconstruction of the anterior surface of the ear using a postauricular pull through neurovascular island flap. AB - The auricular conchal cavity is a shallow structure in the central part of the ear. It is not only 3-dimensional, but it is a gateway to the external ear canal. Many methods have been described for reconstruction of the defect of concha antihelix: split- or full-thickness skin grafts, regional skin, chondrocutaneous and musculocutaneous flaps, but none of the authors have described this flap with neurovascular pedicle. We used postauricular neurovascular pedicle island flap for conchal and periconchal areas of anterior surface of the external ear because it matches to the skin color, thickness and texture; scars are well hidden, and there is no donor-site morbidity. Moreover, it has a constant and reliable neurovascular pedicle. The flap edema which was present in the early postoperative period began to resolve gradually after 3 weeks and disappeared in a few months. The esthetic results were excellent in all of the patients and very satisfying for the patients, too. The sensibility was positive by light touch, pin-prick, temperature and static 2-point discrimination in the postoperative control. The static 2-point discriminations in the transferred flap and in the other ear, which corresponds to the same area, were measured. The results were nearly the same as normal values. We advocate postauricular neurovascular pedicle island flap for conchal and periconchal areas of anterior surface of the external ear because it matches the skin color, thickness and texture; scars are well hidden, and there is no donor site morbidity. Moreover, it has a constant and reliable neurovascular pedicle. PMID- 16721072 TI - The reverse dorsal metacarpal flap: experience with 153 cases. AB - The authors have performed reverse dorsal metacarpal flaps in 153 cases to cover skin defects over fingers or stumps during the past 15 years. Its indications included acute and chronic skin defects over the second to fifth fingers or stumps. The coverage of the flap ranged from the base of the fingers to the distal interphalangeal joint. One hundred forty-three cases survived uneventfully, and 8 cases presented venous congestion postoperatively, which led to epidermal necrosis. The reverse dorsal metacarpal flap is a reliable and excellent method to cover skin defects over the fingers. PMID- 16721073 TI - Clinical experiences with a new tourniquet device. AB - Cessation of blood flow to the extremity improves the comfort of the surgeon. So pneumatic tourniquets are commonly used to obtain a bloodless field during upper- and lower-extremity surgery despite the several problems that tourniquet pressure may lead to. In this paper, we present our clinical experiences with a new tourniquet device called S-MART (OHK Medical Devices, Haifa, Israel) and compared the device with the traditional pneumatic tourniquet in terms of efficacy and complications. A total of 30 patients were included in the study. Preoperative systolic blood pressure measurements were obtained, and patients with a systolic blood pressure >190 mm Hg were excluded. S-MART was used in stead of pneumatic tourniquet in half of the patients in group 1. Pneumatic tourniquet was applied to the patients in group 2. The patient groups were compared. No major complications were observed in either group. But it was observed that S-MART was unsuccessful to maintain a bloodless field during the whole operative procedure in some of the cases. In conclusion, application of S-MART is practical, provides bloodless field for a certain time, and does not increase the complication rate related with the pressure applied to underlying tissues, but it is not a suitable tourniquet device for long surgical procedures. PMID- 16721074 TI - Donor side selection in mandibular reconstruction using a free fibular osteocutaneous flap. AB - While the free fibular osteocutaneous flap is indispensable for mandibular reconstruction, reliable setting is often difficult because relative positions of the bone, skin island, and vascular pedicle are critical. We have an algorithm for donor-side selection of free fibular osteocutaneous flap. From July 2002 to March 2004, we performed 15 mandibular reconstructions using free fibular osteocutaneous flaps. We retrospectively classified these procedures as follows. In type I (flap harvested ipsilaterally to defect, n = 5), the skin island was fixed to the oral cavity, and the vascular pedicle emerged from the anterior aspect of the fibula. In type II (flap contralateral to defect, n = 5), the skin island was fixed to the oral cavity, and the vascular pedicle arose from the posterior aspect. In type III (flap contralateral to defect, n = 4), the skin island was fixed to the facial skin and the vascular pedicle arose from the anterior aspect. In type IV (flap ipsilateral to defect, n = 1), the skin island was fixed to the facial skin and the vascular pedicle arose from the posterior aspect. Flaps took completely except in 1 group II case with partial necrosis. Close attention to geometric characteristics of a free fibular osteocutaneous flap facilitates reconstruction of mandibular defects and selection of donor side. PMID- 16721075 TI - Functional total and subtotal heel reconstruction with free composite osteofasciocutaneous groin flaps of the deep circumflex iliac vessels. AB - Functional and esthetic reconstruction of the bony and tendinous structures with a stable, sensate soft tissue integument after complex posttraumatic defects of the heel is demanding. Cases are rare in the literature and hardly comparable due to their heterogeneity. The reconstructive approach has to consider both patient profile and the reconstructive tree, with free microvascular flaps playing a primary role. The goals are the reconstruction of both osteotendinous structures and slender soft tissue lining for proper shoe fitting for ambulation and mechanical and thermal protection. The flap should be sensate in weightbearing areas to optimize gait and to prevent long-term complications by ulcers. The osteofasciocutaneous deep inferior circumflex artery (DCIA) flap is especially suitable for complex heel defects with subtotal or total loss of the calcaneal bone as all components (iliac bone, groin skin, and fascia lata) can have a wide range of size and shape. We operated on 2 cases with this variable composite flap. One patient had a complete heel defect by war shrapnel. The complete calcaneus, soft heel, and Achilles tendon were reconstructed. The second patient had an empty os calcis after a comminuted fracture and a lateral crush-induced soft tissue defect. In both patients, a stable wound closure, osseous integration, and weightbearing ambulation could be achieved. PMID- 16721076 TI - The clinical importance of variations in the surgical anatomy of the superficial peroneal nerve in the mid-third of the lateral leg. AB - The superficial peroneal nerve (SPN) provides fundamental motor and sensory innervation to the leg and foot. A variety of surgical procedures is performed in the vicinity of this nerve, requiring that the surgeon be familiar with its specific anatomy. We dissected 111 legs to define the anatomic position of the SPN and found that the nerve had 4 distinct variations in location. In 77 (69.4%) specimens, the nerve coursed within the lateral compartment of the leg, while in 18 (16.2%) of the legs, the nerve split and contained branches in both the lateral and anterior compartments. The nerve in 7 (6.3%) legs was found within the intermuscular septum, and in 9 (8.1%) of the specimens, the SPN traveled only within the anterior compartment. These results confirm 4 anatomic variants of the SPN, which will aid surgeons in locating the nerve in the lateral aspect of the leg. PMID- 16721077 TI - Soft-tissue coverage of an open tibial wound in the junction of the middle and distal thirds of the leg with the medial hemisoleus muscle flap. AB - A relatively simple but reliable option for soft-tissue coverage of a less extensive tibial wound in the junction of the middle and distal thirds of the leg has never been determined. In this series, the author reports his clinical experience utilizing the medial hemisoleus muscle flap as a local reconstructive option for management of this unique clinical problem. Over the past 2 years, 14 patients underwent a soft-tissue reconstruction of an open tibial wound (4 x 3 to 10 x 5 cm) in the junction of the middle and distal thirds of the leg with the proximally based medial hemisoleus muscle flap. Only the medial half of the soleus muscle was elevated, with attention to preserving critical perforators from the posterior tibial vessels to the flap while allowing adequate arc of flap rotation to cover the exposed fracture site and hardware. All patients were followed for up to 2 years. Only 1 patient developed insignificant distal flap necrosis and was treated with debridement and flap readvancement. All patients had primary healing of their wounds, reliable soft-tissue coverage, evidenced fracture healing, and good cosmetic outcome during follow-up. Therefore, the medial hemisoleus muscle flap described by the author can be a reliable local option for soft-tissue coverage of a less extensive tibial wound in the junction of the middle and distal thirds of the leg with good outcome and minimal morbidity. PMID- 16721078 TI - Soft-tissue sarcomas and reconstruction options: twenty-two years of experience. AB - Soft tissue sarcomas (STS) are particularly rare malignancies that constitute less than 1% of all malignancies. In recent years, prognostic clinical factors have been defined that help to stratify patients regarding their risk for local and distant recurrence and death from disease. Tumor grade, size, depth, completeness of resection, and presentation status are among the independent prognostic factors. At present, the treatment of these tumors constitutes a wide or marginal excision, adequate primary reconstruction, and radiotherapy. Surgery has generally been recommended as the primary method of treatment for achieving local control. Modern reconstructive surgery, especially musculocutaneous, either pedicle or free flaps, has made more extensive resections possible, while providing acceptable cosmetic and functional results. This study deals with our experience in the treatment of resectable STS with selective combination of treatment modalities. PMID- 16721079 TI - Use of negative-pressure dressings and split-thickness skin grafts following penile shaft reduction and reduction scrotoplasty in the management of penoscrotal elephantiasis. AB - From 1988 to 2005, 8 men who presented with penoscrotal elephantiasis underwent penile shaft degloving and reduction scrotoplasty, followed by transplantation of a split-thickness skin graft (STSG) to the penile shaft. The etiology of elephantiasis in these patients included self-injection of viscous fluid and postsurgical obstructive lymphedema. In the 6 most recent cases, negative pressure dressings were applied over the STSG to promote graft take, and STSG take rate was 100%. The results of our series corroborate those of a previous report, which showed circumferential negative-pressure dressings to be safe and efficacious in bolstering STSGs to the penile shaft. Furthermore, these results suggest that the use of negative-pressure dressings may improve graft take in this patient population. PMID- 16721080 TI - Surgical treatment of axillary hyperhidrosis: a study comparing liposuction cannulas with a suction-curettage cannula. AB - BACKGROUND: Suction-curettage is a minimally invasive surgical approach for the treatment of focal axillary hyperhidrosis. Studies comparing the efficacy of different surgical cannulas are missing. Therefore, we gravimetrically compared a specially designed cannula versus 2 standard liposuction cannulas. METHODS: Axillary suction-curettage was performed in 42 patients (n = 42). Fourteen patients (n = 14) were operated with a 1-hole liposuction cannula, 14 patients (n = 14) with a larger 3-hole liposuction cannula, and 14 (n = 14) with a sharp suction-curettage cannula. Sweat rates in mg/min were measured by gravimetry before and 6 months after surgery. RESULTS: Sweat rate in the 1-hole liposuction cannula group was significantly reduced from 57.65 +/- 5.85 mg/min to 32.58 +/- 4.64 mg/min (P < 0.001), corresponding to a reduction of 44.15%. Patients operated with the larger 3-hole liposuction cannula showed a significant reduction of sweat rate from 63.95 +/- 8.25 mg/min to 33.14 +/- 6.25 mg/min (P < 0.001), corresponding to a reduction of 49.19%. The largest reduction of sweat rates (63.07%) was achieved with the suction-curettage cannula (61.85 +/- 9.03 mg/min to 21.27 +/- 4.42 mg/min (P < 0.001). No severe side effects were observed. CONCLUSION: Due to the higher efficacy we recommend performing suction curettage with a sharp suction-curettage cannula. PMID- 16721081 TI - Bilateral vascularized femoral bone transplant: a new model of vascularized bone marrow transplantation in rats, part I. AB - We present a new model of vascularized bone marrow transplantation-bilateral vascularized femoral bone (BVFB) isograft transplant based on abdominal aorta and inferior vena cava. A total of 7 BVFB isograft transplants were performed between Lewis (RT1) rats. In the donor, both femoral bones were harvested based on the abdominal aorta and inferior vena cava. In the recipient, the harvested isograft transplants were transferred into the inguinal region (in 3 animals) and into the abdominal cavity (in 4 animals). The mean operation time was 3 hours and 35 minutes. The mean warm ischemic time was 35 minutes. The vascular pedicles of the transplants that were transferred into the inguinal region were thrombosed at day 7 posttransplantation. The vascular pedicles of transplants into the abdominal cavity were patent and the bones were viable during the follow-up period of 63 days posttransplant. We have confirmed the feasibility of BVFB transplantation based on abdominal aorta and inferior vena cava. PMID- 16721082 TI - A hemifacial transplantation model in rabbits. PMID- 16721083 TI - Improvement of venous flap survival by application of vascular endothelial growth factor in a rat model. AB - Expression of vascular endothelial growth factor (VEGF) in the venous flap and the effect of exogenous VEGF on survival of the venous flap were studied in rats. A 4- x 4-cm groin type 2 venous skin flap was used in the study. In part 1, biopsies were taken from the flap at 0, 6, 12, 24, and 48 hours after the flaps were raised. VEGF gene expression was measured. In part 2, exogenous VEGF (1 microg/mL) was injected subdermally into the flaps in 10 rats before the flaps were replaced. Flaps that received a saline injection were used as the control. Skin paddle survival was measured on postoperative day 7. The results showed that VEGF expression was significantly increased at 24 and 48 hours after venous flap elevation (P < 0.05). Injection of exogenous VEGF to the flap significantly improved survival of the flap (73% of the flap) when compared with the control, which had a 39% mean percent survival (P < 0.05). We conclude that VEGF expression was increased in the venous flap. Administration of exogenous VEGF significantly improved survival of the venous flap. PMID- 16721084 TI - The establishment of a Pseudomonas aeruginosa-infected burn-wound sepsis model and the effect of imipenem treatment. AB - INTRODUCTION: We present a standardized Pseudomonas aeruginosa (PA) infected burn wound model in mice for evaluating new antimicrobials and therapy strategies for PA infections and demonstrate the effect of the antibiotic imipenem in this model. METHODS: A 6%-8% total body surface area, full-thickness, scald-burn wound was induced in anesthetized mice. Two study groups (PA-infected burn) were compared with 1 treatment group (systemic imipenem) and 3 control groups (noninfected burn, infected nonburned, and burn with distant infection). Seven day mortality, quantitative culture from eschars and from spleens, weight loss, and time to healing were compared. RESULTS: The 25%-100% mortality rate in the nontreated PA-infected burn group was directly related to the infecting inoculum. Imipenem treatment reduced the mortality rate to 0-17%. No control animal died. Systemic bacterial dissemination at 48 hours was significantly higher in the study group. Morbidity paralleled survival results. Wound healing was quicker in the imipenem-treated group and control groups compared with the infected nontreated group. CONCLUSIONS: The mice model is a useful tool for evaluating new antibacterial agents and strategies for treating PA-infected burn injuries. Imipenem was found to be efficacious in the treatment of severe PA sepsis. PMID- 16721085 TI - Definitive closure of the infected median sternotomy wound: a treatment algorithm utilizing vacuum-assisted closure followed by rigid plate fixation. AB - Mediastinitis and sternal wound dehiscence are devastating and life-threatening complications of median sternotomy incision. Ten consecutive patients between July 2001 and May 2005 were diagnosed with sternal wound infection and dehiscence following median sternotomy. Patients were managed by precise debridement and wound excision in the operating room and then dressed with vacuum-assisted closure device. Intravenous antibiotics were prescribed for wound and blood culture microbiological sensitivity. When wounds were bacteriologically controlled, patients returned to the operating room for definitive closure using rigid sternal plating. All patients were extubated postoperatively. No patients died. Average total hospital stay was 21 days. The pectoralis advancement flap was exclusively used for soft tissue reconstruction in 7 patients. There were 2 cases of chronic superficial sternal infection requiring plate removal; however, bony union of the sternum was achieved in all patients. This treatment algorithm provides a useful management strategy for patients with complicated median sternotomy. PMID- 16721086 TI - Correction of late post-traumatic enophthalmos using a tissue expander. AB - We described a new technique to correct late post-traumatic enophthalmos using volume augmentation with a tissue expander. A 47-year-old male requiring enucleation with an implant replacement following orbital fracture with the globe injury had been complaining of persistent enophthalmos and cosmetic defect. Computed tomography scan demonstrated significant enophthalmos resulting from a volume discrepancy between an orbita and the orbital contents. To prevent worsening of the prosthesis motility with correction of enophthalmos, projection of the prosthetic globe was postoperatively adjusted by gradual inflation of an expander placed behind the enucleation implant. As a result enophthalmos was appropriately corrected without any change of the prosthesis motility. PMID- 16721087 TI - The compound dorsal metacarpal artery tendofasciocutaneous flap: an attractive reconstructive tool for complex digital defects. AB - Different modifications of flaps based on the dorsal metacarpal arterial system have been reported as reliable tools for reconstruction of dorsal and palmar phalangeal soft tissue defects. For simultaneous reconstruction of the extensor aponeurosis and joint in complex injuries of the dorsal index, we used a distally based pedicled fasciotendocutaneus second dorsal metacarpal flap, including the complete vascularized extensor indicis tendon in 2 cases. Clinical results of this procedure have not been reported so far. PMID- 16721088 TI - A simple technique for applying vacuum-assisted closure therapy over the circular type external fixation device. AB - Vacuum-assisted closure (VAC) therapy is a widely used method to promote granulation tissue in lower-extremity wounds. However, maintenance of a tight seal over irregular surfaces is difficult, especially around ring fixators. The presented technique in this article offers a time-saving and safe method to achieve an airtight seal around circular fixator in VAC therapy. PMID- 16721090 TI - Gaining entry into plastic surgical training in the United Kingdom: a comparative study with orthopedics and otolaryngology. PMID- 16721091 TI - Ligature technique for superficial veins. PMID- 16721093 TI - Contrary intermittent skin release of complete syndactyly without skin graft in adults. PMID- 16721094 TI - Vacuum-assisted closure garment: method to aid application of a vacuum-assisted dressing. PMID- 16721095 TI - A case of regressive occipital cholesteatoma mimicking an ordinary pilar cyst. PMID- 16721096 TI - Hyperlinking with your tablet personal computer: enhancing your surgical logbook. PMID- 16721097 TI - The major role of a minor plastic surgery operation theater. PMID- 16721098 TI - Diphallus accompanied by anomaly of the urinary system. PMID- 16721101 TI - Fetal growth compromise: definitions, standards, and classification. AB - Fetal growth compromise is mostly defined by gestational age specific threshold value of birth weight or ultrasonically estimated fetal weight. The most frequently used threshold is the 10th centile. However, this approach can not differentiate fetal constitutional smallness from fetal growth failure. The most common descriptor terms are fetal growth restriction (FGR), intrauterine growth restriction and small for gestational age. It is recommended to restrict the use of the first two terms to prenatal assessment of growth and the last term to assessment of the birth weight. Growth reference charts are affected by physiological and methodological variables. There is controversy on whether the reference charts should be customized by multiple variables or inclusive of the whole population. Symmetric FGR is more common and has better outcome than asymmetric FGR. However, such a classification may not be clinically relevant. PMID- 16721102 TI - Fetal growth restriction: pathogenic mechanisms. AB - Complex genetic and environmental mechanisms of maternal, fetal and placental origin regulate fetal growth and may contribute to fetal growth restriction (FGR). The somatotrophic regulatory factors include IGF-I, IGF-II, the IGF binding proteins (IGFBP) 1-6, IGF receptors 1 and 2, and the IGFBP specific proteases. Abnormal remodeling of utero-placental arteries and abnormal fetal placental angiogenesis has also been implicated in FGR. The underlying molecular mediators include vascular endothelial growth factor (VEGF), placental growth factor (PlGF), VEGF receptors, VEGF binding proteins, and numerous other agents working through multiple pathways many of which still remain unknown. Expression of these major angiogenic factors appears to be regulated by local oxygen partial pressure. Future investigations may resolve many of these issues not only adding to the clarity of our understanding of the mechanisms of growth restriction but also improving clinical management. PMID- 16721103 TI - Fetal growth restriction: the etiology. AB - Fetal growth restriction (FGR) is etiologically associated with various maternal, fetal and placental factors, although such an association may not be present in many cases. Maternal factors include hypertensive diseases, autoimmune disorders, certain medications, severe malnutrition, and maternal lifestyle including smoking, alcohol and cocaine use. Fetal etiologies include aneuploidy, malformations, syndromes related to abnormal genomic imprinting, perinatal viral or protozoan infections, preterm birth, and multiple gestation. Placental factors may involve many conditions including anatomical, vascular, chromosomal and morphological abnormalities. Better understanding of these etiologic conditions may lead to improved prediction, prevention and management of FGR. PMID- 16721104 TI - Placenta and fetal growth restriction. AB - The placenta, as the vector for all maternal-fetal oxygen and nutrient exchange, is a principal influence on birthweight. Placental weight summarizes laterally expanding growth of the chorionic disc, and villous arborization yielding the nutrient exchange surface. These different growth dimensions alter fetoplacental weight ratio and ponderal index, and thus may modify placental functional efficiency. The placenta may show a range of histopathologies, some of which are also associated with fetal growth restriction. Different fetal intrinsic abilities to compensate for gross and histo-pathology may clarify the imperfect relationships between fetal growth and both intrauterine pathology, and the long term health risks associated with poor fetal growth. PMID- 16721105 TI - Perinatal outcome and later implications of intrauterine growth restriction. AB - This chapter reviews outcomes for children who have intrauterine growth retardation (IUGR) or small-for-gestation-age (SGA) status at birth. Such infants are at risk for increased perinatal mortality, birth adaptation complications, including perinatal acidosis, hypoglycemia, hypothermia, coagulation abnormalities, and selected immunologic deficiencies. IUGR infants also appear to be at great risk for complications of prematurity, including chronic lung disease and necrotizing enterocolitis. Childhood implications for IUGR include an increased risk for short stature, cognitive delays with decreased academic achievement, and a small but significant increased risk of neurologic disorders, including cerebral palsy. Low socioeconomic status is correlated with the occurrence of IUGR and is significantly related to long-term disabilities. Morbidities associated with preterm delivery appear to be additive to those associated with fetal growth restriction so IUGR, preterm infants may be at great risk for poor neurodevelopmental outcome. PMID- 16721106 TI - Adult consequences of fetal growth restriction. AB - Low birthweight in relation to the length of gestation, is now known to be associated with increased rates of coronary heart disease and the related disorders stroke, hypertension and type 2 diabetes. These associations extend across the whole range of birthweight, which implies that normal variations in nutrient delivery to the fetus have profound long-term effects. The associations are thought to reflect the body's plasticity during development, by which its structure and function can be permanently changed by the intra uterine and early post natal environment. Slow growth during infancy and rapid weight gain after the age of two years exacerbate the effect of slow fetal growth. Cardiovascular disease and type 2 diabetes arise through a series of interactions between environmental influences and the pathways of development that precede them. PMID- 16721107 TI - Screening for fetal growth restriction. AB - Since antenatal detection of fetal growth restriction, defined as birth weight <10% for gestational age, can reduce perinatal morbidity with antepartum testing and use of Doppler, it is imperative that there be a greater effort to detect the growth abnormality. According to a well-conducted randomized clinical trial, all uncomplicated pregnancies should have sonographic assessment of birth weight at 30-32 weeks and at 36-37 weeks. An increased awareness not only of the risk factors but also of the associated probability of abnormal growth can identify the cohorts that would benefit from uterine artery Doppler in 2nd trimester. Among patients at risk for suboptimal growth, Doppler of the umbilical artery improves the detection rate. PMID- 16721108 TI - Sonographic diagnosis of fetal growth restriction. AB - Though a number of ultrasonic methods of diagnosing intrauterine growth restriction (IUGR) exist, the combined use of Doppler velocity wave form analysis of fetal vessels and the ultrasonic estimation of fetal weight (or abdominal circumference) appears to be the best method of both identification and evaluation of IUGR. Once identified, the IUGR fetus should undergo serial evaluation every two weeks or on a weekly schedule (or more frequent interval if the clinical situation warrants) with both Doppler velocity wave form studies and biophysical testing. Determination of the optimal time of delivery depends not only on the results of the antenatal testing, but also on the individual clinical situation. Fetal venous Doppler studies may give additional information about the time frame and significance of the IUGR. PMID- 16721110 TI - Management of fetal growth restriction: an evidence-based approach. AB - This review provides an evidence-based approach to the management of fetal growth restriction (FGR). The management consists of the following components: appropriate fetal surveillance, timely intervention, and selective etiological management. Umbilical arterial (UA) Doppler sonography is the primary test. Supplementary tests include nonstress test (NST), amniotic fluid assessment, biophysical profile (BPP), and selective venous Doppler sonography. Ominous signs include UA absent or reverse end-diastolic flow, non-assuring NST, low BPP, and abnormal fetal venous flow patterns. An evidence-based clinical management guideline is included and individualization of care is recommended. PMID- 16721109 TI - Prenatal therapy for fetal growth restriction. AB - The choice of therapy for fetal growth restriction (FGR) depends on the nature of the insult that led to the development of FGR. Many etiologic factors are either not amenable to therapy or fetal growth has not been improved by treatments that benefit the mother. Many therapeutic approaches have been used to improve fetal condition. None of the approaches have been of value in a consistent manner. We present the numerous approaches, and their rationale for their use, that have been tried to treat the growth impaired fetus. The evidence from the randomized clinical trials is summarized and their conclusions given. PMID- 16721113 TI - Aesthetic body contouring. PMID- 16721112 TI - Aesthetic and functional breast surgery. AB - Approximately 500,000 cosmetic breast operations will be done in 2006. The majority of these are for cosmetic reasons and include breast augmentations, mastopexies, and breast reductions. Augmentations are most commonly performed using saline implants in strictly cosmetic situations and occasionally silicon implants in select cases. Complications are unusual, breast feeding is usually possible and mammographic cancer detection is accurate provided that specific mammographic views are obtained. Most cosmetic breast surgery is not covered by insurance with the exception of breast reduction. PMID- 16721114 TI - Facial rejuvenation. AB - Nonsurgical facial rejuvenation is often used as an alternative to more invasive procedures. Topical agents improve cell maturation and cell turnover, chemical peels and microdermabrasion improve poorly textured skin and pigment anomalies. Surgical facial rejuvenation provides dramatic changes but requires more recovery time. PMID- 16721115 TI - Surgical improvement of the aging forehead and eyelids. AB - Periocular reconstruction is limited to the upper lid, lower lid, and brow. Although many of the same principles apply as in other facial rejuvenation procedures, special techniques are involved frequently using microsurgical procedures. Although the majority of oculoplastic procedures are done for cosmetic purposes, occasionally patients with extreme distortion in periorbital anatomy may benefit from this procedure. These include patients with profound ptosis and severe cases of lower lid distortion who have visual compromise. PMID- 16721117 TI - Laser hair removal. AB - Many techniques are currently available for hair removal. Techniques for permanent hair removal, however, remain elusive. Laser hair removal has become a popular technique in both cosmetic and reconstructive procedures. Laser hair removal can be performed on an outpatient basis. The efficacy of hair removal is difficult to study given the tremendous variation in laser techniques, but short term data is promising. PMID- 16721116 TI - Bariatric surgery. AB - It is estimated that more than 5% of US adults are morbidly obese (body mass index higher than 40). Morbid obesity is associated with adverse health conditions including prolonged morbidity and early mortality while affecting people of lower socioeconomic means. A variety of surgical procedures have evolved over the past 30 years to address this problem with varying degrees of success and longevity. For those patients achieving dramatic weight loss, reconstructive cosmetic surgery is often necessary or desirable. Techniques to achieve this are described. PMID- 16721118 TI - Breast reconstruction. AB - With over 200,000 new cases of invasive breast cancer treated each year, various types of breast reconstruction have become popular and safe. Immediate breast reconstruction seems to confer both benefit and patient satisfaction. A variety of techniques including subcutaneous expanders/implants, flap transfers, and free tissue transfers are described. PMID- 16721119 TI - Current management of varicose veins. AB - Problematic varicose veins are estimated to occur in up to 30 million Americans and may be symptomatic and/or unsightly. The majority of symptomatic varicose veins occur in the lower extremities and can result in significant circulatory disease. This problem is more common in women as a result of pregnancy and child bearing. Cosmetic treatment for varicosities is best achieved with a variety of sclerosing procedures. PMID- 16721122 TI - Side effects of anti-cancer molecular-targeted therapies (not monoclonal antibodies). AB - PURPOSE OF REVIEW: Major advances have been achieved in the field of biologically based therapies for cancer in the last few years, and some of the recently approved 'molecular-targeted therapies' are now being used in daily clinical practice. We aim to review some aspects of the toxicity and safety of small molecule anti-cancer molecular-targeted therapies, with some insights into the physiopathology and predictive factors of toxicity, its correlation with response, and how to prevent and overcome it. RECENT FINDINGS: As a whole, small molecule molecular-targeted therapies are well tolerated. Their toxic profile is favorable, but during the drug development process some severe (sometimes lethal) toxicities have been observed, such as interstitial lung disease in patients treated with drugs targeting the epidermal growth factor receptor. Pharmacogenomic studies can help us to identify those patients with well characterized polymorphisms, and to define the best-tolerated and most effective treatments. SUMMARY: Molecular-targeted therapies have a good toxicity profile in general; however, some patients are exquisitely sensitive to developing particular and severe toxicities related to these drugs. PMID- 16721123 TI - Adverse effects of the humanized antibodies used as cancer therapeutics. AB - PURPOSE OF REVIEW: Humanized monoclonal antibodies represent a recent and very significant addition to the anticancer armamentarium. With improved therapeutic strategies due to these agents, however, there are also various, sometimes unexpected, side effects. RECENT FINDINGS: Most of the monoclonal antibodies used in oncology share a risk of infusion-related manifestations, including the possibility of anaphylaxis; these reactions usually appear early on during the first administration. Hematological toxicity is also frequent, especially if the antibodies are associated with chemotherapy; the resulting neutropenia--and with some agents lymphopenia--is associated with an increased risk of infection. Cardiac failure and pulmonary complications have been reported with some of these agents, especially in patients with prior cardiac or pulmonary comorbidities. SUMMARY: Although consideration of these side effects is important in terms of prevention and therapy, overall they are relatively uncommon, making therapy with monoclonal antibodies quite safe in comparison with other therapeutic modalities used in cancer patients. PMID- 16721124 TI - Neurological complications of cancer chemotherapy. AB - PURPOSE OF REVIEW: To update central and peripheral nervous system neurological manifestations caused by anticancer agents. RECENT FINDINGS: Mostly unpredictable encephalopathy continues to be sporadically reported even in patients treated systemically with conventional chemotherapy doses. Recently, capecitabine, a 5 fluorouracil prodrug, has been added to the list. Magnetic resonance diffusion weighted and fluid-attenuated inversion-recovery imaging are useful in demonstrating chemotherapy-induced central nervous system lesions. The pathogenesis of these lesions is often poorly understood, and is probably multifactorial. A recent observation indicates that genetic polymorphism for methionine is a potent risk factor for methtrexate-induced central nervous system toxicity. Chronic peripheral neuropathy still represents a major limiting factor in a series of chemotherapeutic drugs, and the neuroprotective effect of several older and newer agents is either deceptive or insufficiently proven. In addition to chronic neuropathy, oxaliplatine causes a unique acute syndrome which may respond to calcium plus magnesium infusion. SUMMARY: Neurotoxicity remains a major limitation of many drugs used in cancer patients. Their list grows steadily, and magnetic resonance imaging makes easier the recognition of central nervous system toxicity. Synthesis and thorough clinical testing of neuroprotective molecules remain a major challenge. PMID- 16721125 TI - The prevention of febrile neutropenia. AB - PURPOSE OF REVIEW: Febrile neutropenia causes significant morbidity and mortality in patients receiving cytotoxic chemotherapy. Antibiotic and granulocyte colony stimulating factor prophylaxis reduce the incidence of febrile neutropenia but uncertainty remains regarding their role in clinical practice. We review recent literature to clarify the issue. RECENT FINDINGS: Recent research confirms that prophylactic antibiotics decrease febrile neutropenia and infection-related mortality in acute leukaemia patients and those receiving high dose chemotherapy. Fluoroquinolone prophylaxis also decreases the incidence of febrile neutropenia and all-cause mortality in the first cycle of moderately myelosuppressive chemotherapy for solid tumours. There is no convincing evidence that colonization of individuals with resistant organisms due to antibiotic prophylaxis increases febrile neutropenia or mortality. Granulocyte colony stimulating factor prophylaxis reduces infection-related mortality in patients with greater than 20% risk of febrile neutropenia. SUMMARY: Antibiotic prophylaxis should be offered to patients receiving chemotherapy for acute leukaemia and high dose chemotherapy for solid tumours. It should also be offered to those receiving moderately myelosuppressive chemotherapy for solid tumours and lymphomas during the first cycle of chemotherapy. Prophylactic granulocyte colony stimulating factor is indicated for patients at greater than 20% risk of febrile neutropenia. Further research is indicated to determine whether combining granulocyte colony stimulating factor and antibiotic prophylaxis causes a further reduction in infection-related mortality. PMID- 16721126 TI - Epidermal growth factor receptor/angiogenesis dual targeting: preclinical experience. AB - PURPOSE OF REVIEW: This review aims to critically examine the preclinical background regarding the combination of drugs targeting the epidermal growth factor receptor and anti-angiogenic compounds. RECENT FINDINGS: There are studies exploring the anti-tumor efficacy of dual inhibitors, such as the compound ZD6474, which combines in the same molecule an anti-tyrosine kinase activity against the epidermal growth factor receptor and the vascular endothelial growth factor receptor. In addition, several studies have investigated the anti-tumor effects of combinations of an anti-epidermal growth factor receptor agent and a vascular endothelial growth factor receptor inhibitor. In general, in these studies, supra-additive anti-tumor efficacy was apparent when combining anti epidermal growth factor receptor and anti-angiogenic treatments. Beneficial effects were also observed when combining this dual targeted therapy with either conventional chemotherapy or irradiation. SUMMARY: Early clinical trials combining the anti-epidermal growth factor receptor drug erlotinib (Tarceva) and the anti-angiogenic agent bevacizumab (Avastin) show acceptable toxicity and promising anti-tumor activity (lung cancer), which need to be confirmed in randomized trials. PMID- 16721127 TI - On our way to targeted therapy for cachexia in cancer? AB - PURPOSE OF REVIEW: Cachexia, the occurrence of involuntary weight loss due to loss of adipose tissue and skeletal muscle mass, is among the most common and devastating symptoms in patients with advanced cancer. It is a significant factor contributing to the poor performance status and high mortality rate of these patients, and is a distressing problem for both patients and their families. Despite extensive research in an attempt to better understand the mechanisms involved, progress in the management of cancer cachexia has been slow. RECENT FINDINGS: The pathogenic mechanisms of cachexia and anorexia are multifactorial, but cytokines and tumour-derived factors are known to play a significant role, thereby representing suitable therapeutic targets. Moreover, recent advances in the field of molecular biology have shed light on other mediators involved in the mechanisms leading to muscle wasting, thus increasing potential targets for new therapies. SUMMARY: This review will focus on recent findings in relation to the molecular pathways leading to muscle wasting that have improved our current understanding of cachexia and will direct the future management of cachexia in cancer towards targeted therapies. PMID- 16721128 TI - Dermatofibrosarcoma protuberans: a surgical disease with a molecular savior. AB - PURPOSE OF REVIEW: Dermatofibrosarcoma protuberans is a low-grade malignancy of the skin and subcutaneous tissues with low potential for the development of distant metastases. This tumor is characterized by infiltrative growth, and a chromosomal translocation involving the COL1A1 gene on chromosome 17 and the gene platelet-derived growth factor B gene on chromosome 22. This review will examine recent data confirming the central role of surgery in managing this disease and new findings for the application of molecularly targeted therapy in patients with unresectable disease. RECENT FINDINGS: The largest published series of dermatofibrosarcoma protuberans cases has reported that surgery can lead to excellent local control in dermatofibrosarcoma protuberans in over 95% of cases. In patients with metastatic disease, or with locally advanced disease not suitable for surgical excision, inhibition of platelet-derived growth factor receptors with imatinib (Gleevec) can lead to striking clinical results, including complete responses in some patients. SUMMARY: The identification of platelet-derived growth factor B as a near universal translocation partner in chromosomal rearrangements in dermatofibrosarcoma protuberans has led to the successful application of therapy targeted at platelet-derived growth factor receptors. Although wide surgical excision remains the standard of care, patients with locally advanced disease not suitable for wide surgical excision or with metastatic disease can be managed with the platelet-derived growth factor receptors inhibitor imatinib with high probability of response. PMID- 16721129 TI - ET-743: a novel agent with activity in soft-tissue sarcomas. AB - PURPOSE OF REVIEW: ET-743 (ecteinascidin-743, trabectedin, Yondelis) is a natural marine product that has shown clinical activity in sarcoma. This paper reviews the current knowledge on this compound. RECENT FINDINGS: ET-743 interferes with several transcription factors, traps protein from the nucleotide-excision repair system, thus resulting in DNA damage, modulates gene expression, and blocks cells in the G2-M phase. In the clinical setting, after failure of standard treatment, ET-743 at 1.5 mg/m2 in 24 h continuous infusion every 21 days yielded an overall response rate close to 8% and stabilization rates of 30-40%, some lasting beyond 3 years. Leiomyosarcomas, liposarcomas, and synovial sarcomas may be the more sensitive histotypes. The major toxicities of ET-743 are hepatic--through biliary duct destruction--and hematologic. They are not cumulative and a significant number of patients may receive 12 courses or more. In a randomized Phase II study testing weekly ET-743 with treatment every 3 weeks, an improved progression-free survival rate was observed in the 3-weekly arm; the results of the follow-up Phase III trial should be available at the American Society of Clinical Oncology meeting of 2006. Phase I combination studies are in currently progress. SUMMARY: ET-743 is a novel active drug for sarcoma which yields prolonged disease-free survival in subsets of patients. PMID- 16721131 TI - Mammalian target of rapamycin inhibitors in sarcomas. AB - PURPOSE OF REVIEW: Sarcomas are a rare malignancy accounting for less than 1% of all cancers diagnosed annually. Standard chemotherapy has a response rate of around 25% and newer agents are needed to improve the outcome in patients with advanced sarcomas. The mammalian target of rapamycin plays a central role in cell growth, proliferation, and apoptosis and its inhibition has demonstrated antitumor activity in many tumors and shows promise against sarcomas. RECENT FINDINGS: Recent studies of mammalian target of rapamycin inhibitors in sarcomas have demonstrated clinical benefit response in sarcomas. SUMMARY: Clinical benefit response uses standard Response Evaluation Criteria in Solid Tumors of complete response and partial response as well as stable disease lasting at least 4 months as an endpoint. This endpoint has been shown to select promising new agents against sarcomas. Using this endpoint, the use of the mammalian target of rapamycin inhibitor AP23573 has demonstrated activity against sarcomas. The use of the inhibitor RAD001 (everolimus) along with imatinib in patients with imatinib resistant gastrointestinal stromal tumor has shown promise. Future studies will need to be performed to determine the clinical differences among the mammalian target of rapamycin inhibitors in different subsets of sarcomas. PMID- 16721130 TI - Targeting angiogenesis for the treatment of sarcoma. AB - PURPOSE OF REVIEW: More therapeutic options are needed for bone and soft tissue sarcomas, especially for patients with metastatic disease. Recent randomized clinical trials conducted in colon, breast and lung cancer have shown the anti vascular endothelial growth factor agent, bevacizumab, alone or in combination with chemotherapy, improves response and survival. Preclinical studies have demonstrated the anti-tumor effects of varied anti-angiogenic agents in sarcoma cell lines and tumor models. RECENT FINDINGS: Preclinical studies in sarcomas have evaluated the role of targeted agents including platelet-derived growth factor, matrix metalloproteinases, urokinase receptor and varied small-molecule tyrosine kinase inhibitors. Novel angiogenesis inhibitors are being studied in the treatment of sarcoma, including monoclonal antibodies against vascular endothelial growth factor, cis- and trans-retinoic acids, thalidomide, and tyrosine kinase inhibitors. Phase I, II and III clinical trials continue to evaluate these agents alone, in combinations together and combined with standard chemotherapy. We review herein the preclinical rationale and clinical trial results of anti-angiogenesis therapy in the treatment of soft tissue and bone sarcoma. SUMMARY: Preclinical mechanistic study and clinical trials are continuing in order to evaluate the therapeutic role and ultimately validate the efficacy of the varied anti-angiogenesis agents in soft tissue and bone sarcoma. PMID- 16721132 TI - Future directions for immunotherapeutic intervention against sarcomas. AB - PURPOSE OF REVIEW: To discuss advances in immunotherapy as pertaining to systemic therapy for sarcomas. RECENT FINDINGS: Developments in immunology will have direct relevance to studies in sarcomas and other cancers in the near future. Vaccines employing peptides are the backbone of many studies today due to issues of cost and ease of production, but intact or transfected whole tumor cells or antigen-pulsed or transfected dendritic cells are now being investigated as the immunogenic principle in sarcomas and other cancers. The importance of dendritic cells in generating immune responses is better appreciated than ever, as is the role of CD4+ CD25+ regulatory T cells in mediating immune responses. Enhancing costimulatory signals to T cells using anti-CD152 (CTLA4) and other antibodies or expanding anti-tumor cytotoxic T lymphocytes ex vivo are other means to enhance immunity to sarcoma-specific antigens. SUMMARY: As of 2006, few studies are examining the utility of immunotherapy in sarcomas. However, a detailed understanding of the remarkable mechanics of how an immune response is mounted and how T cell activation and/or proliferation can be halted by the tumor will be central to properly design new studies of immunological agents against sarcomas and other cancers. PMID- 16721134 TI - Microarrays in gastrointestinal cancer: is personalized prediction of response to chemotherapy at hand? AB - PURPOSE OF REVIEW: Molecular profiling has proven to be an invaluable tool in cancer research. Although only in its infancy, microarray technology and gene arrays have led to substantial advances in tumor identification, staging and prediction of response. This review outlines some of the more recent advances in the use of microarrays as a novel means to advance the standard of care for patients with gastrointestinal cancers. RECENT FINDINGS: Recent investigations have shown that gene expression profiles can be used to identify, stage, and guide therapeutic intervention in many gastrointestinal cancers. In cases of unknown primary disease, genetic fingerprints can be used to define the origin of the tumor in the majority of cases. Similarly, gene expression has been shown to allow for more accurate staging of patients with a variety of tumor types. Perhaps most exciting is early data that support the potential for microarray to guide therapeutic intervention by providing specific gene fingerprints which correlate with sensitivity to specific chemotherapy, biologic therapy, or other cancer treatments. SUMMARY: Gene microarrays have become a powerful resource in cancer investigations. Individualized cancer care based on specific gene profiles is on the horizon for patients with gastrointestinal cancers. PMID- 16721133 TI - Utility of positron emission tomography in sarcomas. AB - PURPOSE OF REVIEW: Positron emission tomography using [18F]fluoro-2-deoxy-D glucose has been studied as a tool to help predict the malignant potential of sarcomas, prognosis of patients and response to chemotherapy, and to detect disease recurrence. Recent developments in the use of [18F]fluoro-2-deoxy-D glucose positron emission tomography in the clinical management of patients with suspected or diagnosed sarcomas are presented. RECENT FINDINGS: [18F]fluoro-2 deoxy-D-glucose positron emission tomography should not be used in lieu of histology to diagnose sarcomas, but may aid in biopsy planning. [18F]fluoro-2 deoxy-D-glucose positron emission tomography should not replace magnetic resonance imaging and computed tomography imaging for staging or surveillance. In soft tissue sarcomas, [18F]fluoro-2-deoxy-D-glucose uptake correlates with histologic grade and risk of tumor recurrence. Reduction in [18F]fluoro-2-deoxy-D glucose uptake is an early predictor of histologic response of osteosarcoma, Ewing's and soft tissue sarcomas to chemotherapy, and has correlated with clinical outcomes in Ewing's, soft tissue sarcomas and gastrointestinal stromal tumor. SUMMARY: [18F]fluoro-2-deoxy-D-glucose positron emission tomography will likely play increasingly important prognostic and predictive roles in the management of sarcomas. Available data suggest that positron emission tomography is useful clinically in predicting response to therapy early in the course of treatment for both cytotoxic chemotherapy and kinase inhibitors. Additional study to determine the optimal semiquantitative measure of [18F]fluoro-2-deoxy-D glucose accumulation in sarcoma and standardization of positron emission tomography methods is needed. PMID- 16721135 TI - Adjuvant therapy in gastric and cardia cancer: should we irradiate? AB - PURPOSE OF REVIEW: The purpose of this review is to comment on the current status and the place of the (neo)adjuvant therapy of gastric cancer, and on the standardization of care in this setting. RECENT FINDINGS: The definition of optimal surgery remains controversial in gastric cancer. A recent review by the Dutch Gastric Cancer Group supports the so-called 'over-D1' extended lymphadenectomy, without pancreatectomy and splenectomy, as the optimal procedure, avoiding an increased postoperative mortality. The results from the phase III INT 116 trial should not definitively assign adjuvant chemoradiation as a robust standard of care, mainly due to the lack of optimal surgery in this trial. However, the concept of adjuvant chemoradiation will likely become more and more used, and will influence the design of future studies, reinforced by the incorporation of novel agents. If adjuvant chemotherapy failed to significantly increase survival, the use of perioperative chemotherapy (ECF regimen x 3 pre- and postoperative) was recently reported to improve survival, without affecting postoperative mortality and morbidity; mature results from this large phase III Medical Research Council Adjuvant Gastric Cancer Infusional Chemotherapy trial should be considered as an important step implementing neoadjuvant chemotherapy as a new standard of care. Neoadjuvant therapy of locally advanced tumors also offers an ideal setting to assess new combinations, including cytotoxics, biologics and conformational radiation, coupled with translational research. SUMMARY: Much remains to be done before anticipating an incontestable standard of care in gastric cancer, although the recent phase III trials indicate that multimodality therapy could impact on the prognosis of gastric cancer. PMID- 16721136 TI - New developments in gastrointestinal stromal tumor. AB - PURPOSE OF REVIEW: This review will provide an update of important studies in gastrointestinal stromal tumor with an emphasis on those published over the past 2 years. RECENT FINDINGS: Over the past 60 years basic scientists, pathologists and clinical investigators have studied gastrointestinal stromal tumor with no major advances in patient care until the late 1990s. Discovery at that time of the critical biological role of Kit in gastrointestinal stromal tumor led to the development of one of the most exciting examples of targeted therapy to date. The success of the Kit tyrosine kinase inhibitor, imatinib mesylate (Gleevec, formerly STI-571), has caught the attention of the medical community. With the use of targeted therapy in a targetable disease, new developments in our understanding of epidemiology, genetics, histopathology, radiographic imaging and the biology of gastrointestinal stromal tumor have become apparent. Recent findings are discussed herein. SUMMARY: Continued intense study of gastrointestinal stromal tumor may lead to new paradigms that could revolutionize all of oncology. PMID- 16721138 TI - Endocrine tumors of the pancreas. PMID- 16721137 TI - Bibliography. Current world literature. Supportive care. PMID- 16721140 TI - Issues of computerized tomography scans in children and implications for emergency care. PMID- 16721141 TI - Update on appendicitis: diagnosis and presurgical management. AB - PURPOSE OF REVIEW: Practitioners who see pediatric patients with acute complaints are commonly faced with making the diagnosis of appendicitis. While sometimes the history and physical examination are sufficient, frequently some diagnostic confirmation will be necessary. While diagnostic imaging is frequently useful, it may not be easily obtained. Blood tests are more readily available, with less cost and risk, but unclear diagnostic accuracy. Other questions for the initial examiner are when to consult the surgeon and if analgesia administration will obscure the diagnosis. RECENT FINDINGS: Abdominal computed tomography scans and ultrasound scans have been shown to be accurate in the diagnosis of acute appendicitis. The literature has begun to focus on which patients should undergo imaging studies. Several clinical decision rules that may be helpful in identifying low-risk patients have been derived. Analgesia given before establishing a diagnosis does not appear to change outcomes. SUMMARY: A careful history and physical examination will identify a minority of pediatric appendicitis cases. When diagnostic testing is needed, computed tomography scanning has the best test performance, but also carries the risk of radiation. Elimination of a portion of low-risk patients may reduce unnecessary imaging. PMID- 16721142 TI - Update on the acute management of status epilepticus in children. AB - PURPOSE OF REVIEW: Status epilepticus is the most common neurologic emergency in children. The understanding of its less recognizable forms, its pharmacologic management, the role of electroencephalography and the long-term morbidity and mortality as a result of status epilepticus are consistently evolving. This review frames the current understanding of several issues as they apply to acute management in the emergency department. RECENT FINDINGS: Researchers are working to define less recognizable forms of status epilepticus such as nonconvulsive, autonomic and psychogenic. Buccal and intranasal forms of midazolam are emerging as suitable alternatives to rectal diazepam in the initial treatment of status epilepticus. Valproic acid, chloral hydrate and newer-generation antiepileptics are being proposed as safe and effective alternatives to the traditional drugs used to treat status epilepticus. The role of electroencephalography in diagnosis is being elucidated. Risk factors for neurologic sequelae and mortality after status epilepticus remain an area of research with conflicting findings and no real consensus. SUMMARY: The understanding of different types of status epilepticus, the options for pharmacologic treatment, the tools for diagnosis and the morbidity and mortality of the disease are still evolving. As a result, several areas for further research remain that will help clinicians in their approach to this complex condition. PMID- 16721143 TI - Advances in the emergency management of pediatric sepsis. AB - PURPOSE OF REVIEW: Sepsis requires prompt recognition and aggressive therapy; early goal-directed therapy decreases morbidity and mortality. Recommendations on the specific management of pediatric sepsis have historically been extrapolated from adult literature and from expert/consensus opinion. This review serves to appraise recent recommendations and determine the applicability of newly promoted adult guidelines for pediatric sepsis. RECENT FINDINGS: The first and most recent recommendations on the management of pediatric sepsis were published in 2002 by expert consensus with the recognition that there were few data from randomized controlled trials or meta-analysis. Previous to this, adult guidelines were used in the management of pediatric sepsis. New definitions for pediatric sepsis provided by expert consensus take into account the specific physiologic changes. Prompt correction of hemodynamic derangements has been shown to improve outcome through aggressive volume resuscitation and early initiation of vasopressor therapy. Frequent reassessment has been emphasized to ensure appropriate management. SUMMARY: Prompt recognition, institution of appropriate therapy, and continual assessment for children with sepsis and shock are essential for improved outcomes. The emerging new literature on early goal-directed therapy is most applicable to care in the emergency department. PMID- 16721144 TI - Update on the management of skin, soft-tissue, and osteoarticular infections in children. AB - PURPOSE OF REVIEW: To provide an update on the diagnosis and management of skin, soft-tissue, and osteoarticular infections in children. RECENT FINDINGS: Our understanding of the epidemiology of skin and soft-tissue infections and osteoarticular infections is changing rapidly. Community-associated methicillin resistant Staphylococcus aureus has become a predominant cause of childhood skin and soft-tissue infections. Kingella kingae is also increasingly identified as a cause of osteoarticular infections. Challenges in Staphylococcus aureus treatment and Kingella kingae identification are changing the approach to skin and soft tissue infections and osteoarticular infections. SUMMARY: Community-associated methicillin-resistant Staphylococcus aureus should be considered a cause of skin and soft-tissue infections. Empiric antimicrobial choices should be modified in areas in which there is a more than 10% prevalence of community-associated methicillin-resistant Staphylococcus aureus infection. Decontamination of shared sports equipment should be undertaken to minimize person-to-person spread. No established guideline for eradication of carriage of community-associated methicillin-resistant Staphylococcus aureus exists. Kingella kingae is a more prevalent cause of osteoarticular infections than previously recognized and can cause outbreaks of invasive infection via person-to-person transmission. Modification of culturing procedures for osteoarticular infections including inoculation of infected joint fluid and bone in blood-culture bottles should be considered. PMID- 16721145 TI - All-terrain vehicle, trampoline and scooter injuries and their prevention in children. AB - PURPOSE: Childhood injuries are the leading cause of death in children and result in significant healthcare utilization. Injuries specifically related to all terrain vehicles, trampolines and scooter usage can be devastating and are often preventable. Our understanding of how and why these injuries occur can aid in preventing morbidity and mortality. RECENT FINDINGS: The popularity of all terrain vehicles, nonmotorized scooters and trampolines has soared over recent years. This increased usage has led to a tremendous rise in injuries in children utilizing these recreational activities. Many of the injuries occur in younger children who may not possess the motor and cognitive skills necessary to safely engage in these activities. These activities lead to a number of head and extremity injuries, most of which can be attenuated by the use of protective gear such as helmets and protective padding. SUMMARY: Understanding the nature of these injuries can lead to advocacy and hopefully legislation to prevent further injuries from occurring. PMID- 16721146 TI - Emergency and critical care pediatrics: use of medical simulation for training in acute pediatric emergencies. AB - PURPOSE OF THE REVIEW: Recent data suggest that pediatric trainees receive insufficient training to manage acute pediatric emergencies. This review addresses the use of medical simulation as a way for medical learners to acquire and maintain skills needed to manage pediatric resuscitations. RECENT FINDINGS: Recent work highlights the value of deliberate practice in the context of simulated medical environments to promote development of medical expertise. Several studies demonstrate the benefit of simulation-based training for improved skill acquisition in advanced life support, emergency airway management, and nontechnical skills. Work in other fields of medicine supports the integration of simulation into pediatric training programs. SUMMARY: Medical simulation holds great promise to enhance existing pediatric training curricula by increasing skills and expertise in resuscitation. Future research is needed to identify best methods of pediatric simulation-based training. PMID- 16721148 TI - Airway clearance in children with neuromuscular weakness. AB - PURPOSE OF REVIEW: Assistance with airway clearance is critical for reducing morbidity and mortality in children with neuromuscular weakness. Several techniques and devices are available to enhance airway clearance in patients with neuromuscular disease. Only recently, however, has assessment of their effectiveness included children. This review highlights the rationale for use of both secretion extraction and mucus mobilization techniques and devices, emphasizing findings in pediatric patients whenever possible. RECENT FINDINGS: Cough in adults is adequate when peak cough flow exceeds 160 l/min. Similar threshold values in young children have not been established. Those methods that enhance secretion extraction include breath stacking, manual or mechanical insufflation, manually assisted cough, and mechanical insufflation-exsufflation. All are well tolerated in children and can increase peak cough flows, although the greatest increase occurs with mechanical insufflation-exsufflation. Techniques successfully used to help mobilize secretions include high frequency chest wall oscillation and intrapulmonary percussive ventilation. Various modalities can be used successfully alone or in combination. SUMMARY: Secretion extraction and mobilization techniques are safe, even in infants who require airway clearance assistance. To date, however, criteria specific for children are lacking to determine when such modalities should be used and which ones are most effective. PMID- 16721147 TI - Ventilatory management of sleep-disordered breathing in children. AB - PURPOSE OF REVIEW: The purpose of this review is to summarize the current management of continuous positive airway pressure and noninvasive positive pressure ventilation in children with sleep-disordered breathing. RECENT FINDINGS: Although most children with sleep-disordered breathing respond to surgical treatment, the use of continuous positive airway pressure and noninvasive positive pressure ventilation in nonresponders has become common, with hundreds of cases reported in the literature, despite the fact that these devices are not approved by the Food and Drug Administration for use in children weighing under 30 kg. Studies show that continuous positive airway pressure and non-invasive positive pressure ventilation are safe and efficacious. Side-effects are minor, and include nasal symptoms and skin breakdown. Midfacial hypoplasia is an uncommon adverse event. Problems with triggering and cycling of noninvasive positive pressure ventilation remain an issue in small or weak children. As in adults, poor adherence is the major obstacle to successful continuous positive airway pressure or noninvasive positive pressure ventilation use. SUMMARY: Continuous positive airway pressure is a useful second-line treatment for children with sleep-disordered breathing. Strategies to improve adherence are needed. Equipment manufacturers should be encouraged to develop equipment that better meets children's needs. PMID- 16721149 TI - Current concepts in surgical management of pediatric lung and chest wall diseases. AB - PURPOSE OF REVIEW: The management of pediatric lung and chest wall diseases has changed dramatically in the last decade because of the application of minimally invasive surgical techniques. This review will try to highlight some of the more significant recent contributions. RECENT FINDINGS: Most of the papers quoted will show that the application of minimally invasive surgery to the treatment of common problems such as empyema, pneumothorax, and lung biopsy has significantly altered our treatment algorithms, because of decreased morbidity and improved outcomes compared to standard surgical or medical treatment plans. This has also been true for pediatric chest wall deformities, which previously were treated in only the most severe cases and thought to be mostly cosmetic in nature. More recent reports now show statistically significant improvements in cardio pulmonary physiology as well as quality of life. SUMMARY: The application of less invasive surgical procedures for lung and chest wall diseases has warranted earlier intervention, with better outcomes, and less morbidity than previously used techniques. A clear understanding of these techniques and their benefits is important for the referring and treating physician as previously used treatment protocols may no longer provide the best patient care. PMID- 16721150 TI - Genetics of pediatric interstitial lung disease. AB - PURPOSE OF REVIEW: The interstitial lung diseases are a heterogeneous group of rare disorders of largely unknown etiology. The occurrence of familial cases of pediatric interstitial lung diseases with the onset of symptoms developing early in infancy has suggested a genetic basis for some forms of interstitial lung diseases in children. RECENT FINDINGS: Mutations in the genes encoding surfactant protein C, SFTPC, and a member of the adenosine triphosphate-binding cassette family of proteins, ABCA3, have been shown to result in pediatric interstitial lung diseases inherited in autosomal-dominant and autosomal-recessive patterns, respectively. There is overlap in both the clinical and histopathologic features of these disorders. SUMMARY: Identification of genes responsible for pediatric interstitial lung diseases provides the opportunity for noninvasive testing to establish an etiologic diagnosis, to counsel family members for their recurrence risk, and to classify these rare disorders more accurately. A better understanding of the cause and pathophysiology of these disorders may provide additional insights into the causes of other forms of pediatric interstitial lung diseases, and may suggest novel treatment approaches. PMID- 16721152 TI - Heterotaxia syndromes and their abdominal manifestations. AB - PURPOSE OF REVIEW: Children with heterotaxia often have abnormalities of intestinal rotation, some of which may predispose to midgut volvulus. This review summarizes the literature and proposes an individualized approach to the patient based on symptoms and findings on gastrointestinal imaging. RECENT FINDINGS: This report reviews the types of heterotaxia syndromes and the debate surrounding the optimal management of the associated intestinal rotational abnormalities. Recent publications suggest that not all rotational abnormalities predispose to volvulus and that the natural history of rotational abnormalities in asymptomatic patients with heterotaxia is relatively benign. On the basis of these findings, an algorithm for the management of intestinal rotational abnormalities is proposed. SUMMARY: Malrotation and midgut volvulus is a life-threatening complication. Children with heterotaxia and symptoms suggestive of a proximal intestinal obstruction require urgent investigation and surgical treatment. Asymptomatic children require close clinical follow-up and urgent investigation of gastrointestinal symptoms. PMID- 16721153 TI - Trisomy 21 and the pediatric surgeon. AB - PURPOSE OF REVIEW: Trisomy 21 (Down syndrome) is a common condition encountered by the pediatric surgeon. The surgical anomalies associated with Down syndrome now have treatment options that were previously considered futile. An effective surgical team approach to these children involves knowledge of the unique pathophysiology of trisomy 21 and an appreciation for the desires of the family and the needs of the child. RECENT FINDINGS: Surgical techniques benefit children with Down syndrome equally as typical children. Pessimism regarding the outcome of surgical treatment of colorectal anomalies is fading, with evidence of nearly equivalent outcomes. Several autoimmune disorders impact children with Down syndrome. Idiosyncratic susceptibility to infections and malignancy is noted. Unique anesthetic complications and aerodigestive tract disorders have been identified. Many anomalies have surgical options that enhance the quality of life. SUMMARY: There is optimism in the surgical treatment of children with Down syndrome. Improved operations, better anesthetic management and recognition of problems specific to the child with trisomy 21 make it safer and ethical to offer surgical solutions. Societal acceptance and mainstream participation are on the rise. The astute, compassionate pediatric surgeon does much to enhance the quality of life of these children and of the families who love them. PMID- 16721154 TI - Intersex genetic anomalies with malignant potential. AB - PURPOSE OF REVIEW: Mutations in genes contributing to sexual determination and differentiation can cause clinical syndromes with potential for the development of malignant tumors. This article focuses on intersex disorders requiring surveillance for and/or operation to prevent or treat malignancies. RECENT FINDINGS: Classification of intersex disorders into risk groups gives guidance to physicians about children who are vulnerable to malignant degeneration of the gonads or kidneys. The gonads most at risk are both dysgenetic and intra abdominal, and early gonadectomies are recommended as malignancies have been reported in infancy. Predominant risk groups include syndromes of gonadal dysgenesis and Ullrich-Turner syndrome. Partial gonadectomy is feasible in true hermaphrodites commensurate with sex of rearing. Histologically normal intra abdominal gonads may be left through puberty (androgen insensitivity syndromes). A palpably normal descended gonad in a child with a Y chromosome can be observed if the child is reared as male. Certain intersex syndromes with splice variants of the WT1 gene are susceptible to Wilms' tumors (Frasier and Denys-Drash syndromes). SUMMARY: Prevention or early recognition of malignancy in intersex disorders requires knowledge of the risk factors including dysgenetic gonads, a Y chromosome with intra-abdominal gonads and dysgenetic syndromes with WT1 gene splice variants. This paper describes the evolution toward laparoscopic gonadectomy in intersex patients, as a means to remove abnormal gonads and associated ductal structures as dictated by the disease or syndrome. PMID- 16721156 TI - Intestinal polyposis syndromes: progress in understanding and treatment. AB - PURPOSE OF REVIEW: Dramatic advances in the understanding of the genetic basis for inherited diseases and in diagnosis and treatment are particularly applicable to intestinal polyposis. Recent contributions have impacted our understanding of the molecular basis for the disease and improved our ability to treat them. RECENT FINDINGS: Insights into the genetic basis of the spectrum of disease and refinement of genetic screening, diagnostic tests, and surgical management of intestinal polyposis and extraintestinal manifestations are reviewed. SUMMARY: Our understanding of the genetic basis for the intestinal polyposis syndromes remains incomplete, but recent studies have contributed to filling in significant gaps in our knowledge. Specific genetic alterations have been identified in the majority of the 30% of patients with familial adenomatous polyposis that do not test positive for mutations in the APC gene with routine testing. A novel gene ENG has been identified in a subgroup of patients with Peutz-Jegher's syndrome and a hypothesis has been proposed to explain the pathogenesis of the mucosal defects. Technological advancements in diagnosis and treatment include the use of capsule endoscopy for screening and the minimally invasive approach to total colectomy with ileoanal pullthrough. Treatment of desmoid tumors remains a challenge, but new chemotherapy regimens show some promise in treating this rare, but devastating, extraintestinal manifestation. PMID- 16721155 TI - Surgical intervention in children with multiple endocrine neoplasia type 2. AB - PURPOSE OF REVIEW: We provide a summary of the literature published in the past year addressing the surgical approach to multiple endocrine neoplasia type 2 in the pediatric population. RECENT FINDINGS: The review focuses first on medullary thyroid carcinoma and performing prophylactic thyroidectomy for the prevention or cure of this disease. The timing and extent of surgery as well as additional surgical intervention for persistent or recurrent disease is discussed. Then the surgical management of hereditary pheochromocytoma is reviewed. SUMMARY: Surgery is often the only treatment that can prevent or cure the endocrinopathies associated with multiple endocrine neoplasia type 2. Determining the proper timing and extent of surgical intervention in children affected with multiple endocrine neoplasia type 2 will lead to better outcomes and survival. PMID- 16721158 TI - Immunizations, neonatal jaundice and animal-induced injuries. AB - PURPOSE OF REVIEW: To review the literature published within the last year on three topics essential to clinical pediatrics: immunizations, neonatal jaundice, and animal-induced injuries. RECENT FINDINGS: New vaccines that protect against meningococcus, pertussis and rotavirus are safe, effective and recommended for routine immunization. Young children remain a high priority for influenza vaccination while the world awaits further developments of avian influenza. Pneumococcal and varicella vaccinations have benefited many. Debate exists on how to screen for hyperbilirubinemia in neonates and new strategies are emerging to prevent it. There seems to be no link between hyperbilirubinemia and autism spectrum disorders. We have learned that rabies can be transmitted by transplantation; it remains a global public health problem and its incidence is frequently underestimated in developing nations. Lastly, brown recluse spider bites are often misdiagnosed. SUMMARY: The face of pediatric infectious disease is changing as we incorporate new vaccines into our routine practice. Rotavirus vaccine has significant implications for the health of children across the globe. The management strategy for neonatal jaundice continues to focus on screening and prevention. We need to devote more energy to combating rabies in countries where it is endemic. PMID- 16721157 TI - Surgical management of pulmonary and gastrointestinal complications in children with cystic fibrosis. AB - PURPOSE OF REVIEW: Cystic fibrosis is a common disorder, affecting as many as 1:2500 Caucasian live births. Despite improved medical management, disease specific complications are common and are responsible for substantial morbidity and ultimately mortality. Both pulmonary and gastrointestinal complications of cystic fibrosis are well known; however, the complications requiring surgical intervention in the pediatric population are infrequent. We provide a detailed review of the cystic fibrosis-associated pulmonary and gastrointestinal complications and potential surgical options for management in children with cystic fibrosis. RECENT FINDINGS: Recent operative approaches are described that include application of minimally invasive surgical techniques primarily for intrathoracic disease. Novel medical therapies are also presented. Finally an attempt is made to put in perspective those surgical care advances that have had a benefit on disease outcomes. SUMMARY: This report will provide the physician caring for the child with cystic fibrosis an understanding of those disease complications that will require surgical consultation and potential operative intervention. PMID- 16721159 TI - Bibliography. Current world literature. Emergency and critical care medicine. PMID- 16721161 TI - The psychosocial consequences for children and young people who are exposed to terrorism, war, conflict and natural disasters. PMID- 16721162 TI - Literacy and mental disorders. AB - PURPOSE OF REVIEW: This review examines recent evidence on the comorbidity between literacy problems and psychiatric disorder in childhood and discusses possible contributory factors. RECENT FINDINGS: Recent studies confirm the substantial overlap of literacy problems with a range of emotional/behavioural difficulties in childhood. Literacy problems and inattention may share genetic influences, contributing to associations with attention deficit hyperactivity disorder. To an extent, links with conduct problems may be also mediated by attentional difficulties. In addition, findings suggest bidirectional influences whereby disruptive behaviours impede reading progress and reading failure exacerbates risk for behaviour problems. Associations between literacy problems and anxiety disorders are not entirely mediated by inattentiveness. Rather, comorbid anxiety disorders seem likely to arise from the stressors associated with reading failure. Findings in relation to depression are less consistent, but suggest that poor readers may be vulnerable to low mood. Children with autism seem more likely to face problems in reading comprehension than the decoding difficulties more prominent in other disorders. SUMMARY: Literacy problems are associated with increased risks of both externalizing and internalizing disorders in childhood, with different mechanisms likely to be implicated in each case. When comorbid problems occur, each is likely to require separate treatment. PMID- 16721163 TI - Mental health, educational and social needs of young offenders in custody and in the community. AB - PURPOSE OF REVIEW: The purpose of this review is to highlight recent findings from research on mental health, educational and social needs of young offenders and the implications for key agencies that commission or provide services. RECENT FINDINGS: Young offenders have high levels of morbidity in a number of areas, including mental health. A substantial number of young offenders have learning disabilities, which has implications for educational provision and delivering interventions. Other learning problems, such as low reading and comprehension ages, also have consequences for society in a number of areas, including occupational functioning and the associated development of a skilled workforce. There is some evidence that female offenders and persistent offenders are particularly at risk of psychosocial problems. Need is often unmet due to the lack of adequate assessment. Other reasons include inaccessibility of services and difficulties experienced in engaging young people in treatment. Young offenders in the community and those from ethnic minorities are particularly at risk of having poor access to services. SUMMARY: Providing services for young offenders requires investment from all the agencies in prioritising their needs. Research can continue to contribute by informing policy and practice, although further understanding of the developmental trajectories of high-risk groups is still needed. PMID- 16721164 TI - Healthcare issues for children on the autism spectrum. AB - PURPOSE OF REVIEW: We provide a summary of issues relevant to healthcare and health delivery in autism and related disorders, with a focus on work published in the past 5 years. RECENT FINDINGS: Autism has a strong genetic basis as evidenced by the high recurrence rate in families and its associations with Fragile X syndrome and tuberous sclerosis. Early diagnosis of autism is important, given the potential for improvement with intervention. Although its associations have been reported with events such as measles-mumps-rubella immunization, large-scale studies have not supported such links. Controlled drug trials have increased in frequency and drug treatment may be helpful in addressing frequently associated behavioural and emotional difficulties. Alternative treatments are frequently preferred by parents. SUMMARY: Knowledge of autism among healthcare professionals has increased but gaps remain. Physicians and other professionals can facilitate the delivery of healthcare services to individuals with autism. Awareness of empirically validated treatments will facilitate work with individuals with autism and related disorders as well as with their families. PMID- 16721165 TI - Strengths and Difficulties Questionnaire: research and clinical applications. AB - PURPOSE OF REVIEW: To critically discuss recent findings from studies using the Strengths and Difficulties Questionnaire, in relation to different research designs and clinical purposes. RECENT FINDINGS: A substantive body of research exists on the psychometric properties of the Strengths and Difficulties Questionnaire in different cultures. This body supports the validity and reliability of its versions for the parent, teacher and self-reporting purpose, despite some variation in cut-off scores. The Strengths and Difficulties Questionnaire is increasingly being used as a measure of child psychopathology in other types of research, that is, aetiological, longitudinal and service evaluation studies. Future studies need to address in more detail the Strengths and Difficulties Questionnaire diagnostic algorithms, added value score, impact and burden items and a preschool version. SUMMARY: The Strengths and Difficulties Questionnaire has been established as the most widely used instrument in child mental health research. It is easy to complete, is user friendly because of its positive attributes items, allows comparisons to be made between different populations and is sensitive to change. PMID- 16721166 TI - Recent advances in the development of screening tools for mental health in young offenders. AB - PURPOSE OF REVIEW: To describe recent advances in the development of mental health needs screening tools for use with young offenders and implications for this vulnerable group. RECENT FINDINGS: The literature shows that young offenders in North America and Europe have high levels of diverse needs that often go unmet because of lack of methods for recognizing and assessing them. Effective screening could play an important role in improving recognition of these needs. Substantial developments have been occurring in the United States, England and Wales and the Netherlands in developing screening tools for this purpose and for this population. Now, the literature is beginning to provide a broad understanding of the optimal psychometric properties, utility and limitations of the screening tools that are required. SUMMARY: Several centres are engaged in work to construct mental health screening and assessment tools for use with young offenders. Psychometric measures are being used to determine their validity and reliability. The work has highlighted deficiencies in the healthcare of young persons involved in criminal justice systems but should assist policymakers, healthcare strategists, planners and commissioners to come to better informed decisions about how to utilize most appropriately the limited mental health expertise that is available and how to enhance awareness of nonhealth professionals of basic mental health risks, problems and disorders. PMID- 16721167 TI - Services research and outcomes: an introduction. PMID- 16721168 TI - The community psychiatrist of the future. AB - PURPOSE OF REVIEW: The potential contributions of the community psychiatrist are described, via the interdisciplinary team, to individuals and families dealing with mental illness, and to the communities of the future, along with the opportunities for, and barriers to, effective teamwork and community collaborations. RECENT FINDINGS: Health and medical training systems still provide perverse incentives deterring psychiatrists from becoming adequately trained in community psychiatry and full members of interdisciplinary teams, and skilled partners in improving the mental health of the whole community. Sources of potential role conflict should be resolved, and advantages of community collaborations, interdisciplinary leadership, support of teamwork, division of labour, cross-fertilization and hybrid vigour should be realized. SUMMARY: Truly essential and desirable roles and the skill base of community psychiatrists in interdisciplinary teams and local communities could be developed and strengthened by changes in basic and advanced psychiatric training, and by psychiatric professional bodies and training programmes placing greater emphasis and value upon the roles of a community psychiatrist. PMID- 16721169 TI - Incidence, prevalence and mortality of anorexia nervosa and other eating disorders. AB - PURPOSE OF REVIEW: The purpose of this review is to evaluate the recent literature on the incidence and prevalence of and mortality associated with eating disorders. RECENT FINDINGS: General-practice studies show that the overall incidence rates of anorexia nervosa remained stable during the 1990s, compared with the 1980s. Some evidence suggests that the occurrence of bulimia nervosa is decreasing. Anorexia nervosa is a common disorder among young white females, but is extremely rare among black females. Recent studies confirm previous findings of the high mortality rate within the anorexia nervosa population. SUMMARY: The incidence of anorexia nervosa is around eight per 100,000 persons per year. An upward trend has been observed in the incidence of anorexia nervosa in the past century till the 1970s. The most substantial increase was among females aged 15 24 years, for whom a significant increase was observed from 1935 to 1999. The average prevalence rates for anorexia nervosa and bulimia nervosa among young females are 0.3 and 1%, respectively. Only a minority of people with eating disorders, especially with bulimia nervosa, are treated in mental healthcare. PMID- 16721170 TI - The epidemiology and characteristics of stalking. AB - PURPOSE OF REVIEW: This article summarizes major results in the field of epidemiology and characteristics of stalking, as published in literature between 2003 and October 2005. RECENT FINDINGS: Although stalking had been only recently conceptualized, it soon became evident that it represents a significant social and medical problem. Community-based studies on the prevalence of stalking are scarce and predominantly from English-speaking countries. All epidemiological studies, however, revealed that stalking is a widespread phenomenon with lifetime prevalence rates of stalking victimization ranging 12-16% among women and 4-7% among men. With regard to the impact of stalking, studies suggest that stalking can cause serious economic, social, medical and psychiatric consequences. Growing evidence suggests that serious violence and even homicide may occur in the context of stalking, and research on risk assessment has yielded an improved understanding of risk factors. Research on therapy of stalkers and stalking victims is still at its beginning; however, there are some preliminary reports with encouraging results. SUMMARY: Although there are research activities in the main fields, many basic questions still remain to be adequately addressed. Increased social and political awareness and expanded research funding are obligatory prerequisites to realize sound and well designed studies. PMID- 16721171 TI - Management of complex and severe personality disorders in community mental health services. AB - PURPOSE OF REVIEW: To describe the prevalence and specific management of severe and complex personality disorders in community mental health services and similar settings. RECENT FINDINGS: Conflicting reports exist in the literature but the consensus of evidence suggests that more complex personality disorder (more so than simple personality disorder) impairs response to treatment of most psychiatric disorders and complicates their management. Poor adherence to treatment may be one of the mechanisms underlying this. SUMMARY: Increasing evidence now suggests that more complex and severe personality disorders have a negative impact on the outcome of most psychiatric disorders. Some indications, however, are there to show that pharmacological treatments may not handicap the outcome of pharmacological treatment as much as other types of intervention and may be selectively chosen in this group. Severe personality disorder needs to be identified early in community services as by anticipating and adjusting to its effects it is likely to improve treatment plans and prognosis. PMID- 16721172 TI - Epidemiological impact of antidepressant and antipsychotic drugs on the general population. AB - PURPOSE OF REVIEW: To analyse the prevalence of and sex and age distribution associated with antidepressant and antipsychotic drug exposure in the general population and to highlight recent epidemiological findings concerning specific adverse outcomes associated with drug exposure. RECENT FINDINGS: Epidemiological studies indicate high rates of second-generation antidepressant and antipsychotic drug use in the general population. The use is more prevalent among women than among men, and in older rather than in younger age groups. A new pattern of adverse outcomes has been described in individuals exposed to newer agents, including a possible risk of suicidal acts in adults receiving second-generation antidepressants, the risk of cerebrovascular events in older individuals receiving second-generation antipsychotics and the risk of metabolic disturbances in individuals exposed to specific second-generation antipsychotics. SUMMARY: The assessment of, and attention to, the development of specific adverse reactions in individuals exposed to second-generation psychotropic drugs may improve treatment outcomes. PMID- 16721173 TI - Improvement of the physical health of people with mental illness. AB - PURPOSE OF REVIEW: Although excess rates of medical comorbidities in people with mental disorders have been known for years, concrete steps undertaken to fight this unsatisfactory situation have not been undertaken yet. Therefore, in this issue of Current Opinion in Psychiatry, recent findings in five major areas are summarized. RECENT FINDINGS: All major psychiatric disorders seem to be affected with physical comorbidities, but the issue has attracted attention only recently. SUMMARY: Reasons for the excess rates of physical comorbidities in mental illnesses are diverse. They are in part inherent in psychiatric disorders, are related to the problems of the health system and have to do with stigma and the insufficient training of psychiatrists. PMID- 16721174 TI - Somatoform disorders in neurological practice. AB - PURPOSE OF REVIEW: Patients with medically unexplained symptoms continue to intrigue, fascinate and frustrate clinicians. They are common in general medicine and often present with apparent neurological disorder. This review aims to provide insight into the recent literature that has sought to clarify epidemiology, diagnostic issues, aetiologic understanding and treatment of patients with psychogenic disorders who usually first present to neurologists. RECENT FINDINGS: Somatoform disorders are common in neurological practice. A number of papers have addressed issues of epidemiology and identified that medically unexplained symptoms in neurological populations are higher than originally thought. A number of recent review papers have served to summarize areas of considerable information (e.g. treatments) and areas of rapid growth in knowledge (e.g. neuroimaging). Studies investigating the role of psychological factors are well represented and clarify our psychopathological understanding of somatoform disorders in patients presenting to neurologists. Treatment studies are few and continue to be limited by population sizes and study designs. SUMMARY: Somatoform disorders are common in neurological populations. Comorbidity related to somatoform disorders with known organic neurological conditions requires further study. On account of the limitations of treatment studies, evidence-based clinical management of these patients is awaited. PMID- 16721175 TI - Mood disorders: cardiovascular and diabetes comorbidity. AB - PURPOSE OF REVIEW: Depression is often associated with medical comorbidity. New research quantifies patterns of mood disorder in illnesses such as cardiovascular disease and diabetes, evaluates the prognostic significance of mood symptoms, and seeks to identify common mechanisms for both mood and medical disease. This review provides recent findings on comorbidity, summarizes mechanistic hypotheses, and outlines developments in treatment and services. RECENT FINDINGS: Depression occurs in up to one-quarter of patients with cardiovascular disease and diabetes. Depressed patients with heart disease have poorer medical outcomes including increased risk of reinfarction and all-cause mortality. Patients with diabetes and depression have poorer glycemic control, more diabetes symptoms, and greater all-cause mortality. Depression is associated with both biological (hypothalamic-pituitary-adrenal axis dysregulation) and psychosocial processes (adherence, poorer diet, and exercise) that may mediate adverse medical outcomes. Antidepressant treatments are effective in treating depression in medically ill patients, but their impact on medical outcomes remains to be quantified. SUMMARY: Depression, cardiovascular disease, and diabetes are among the most common chronic illnesses affecting an aging population. Depression is treatable in patients with medical illnesses, and collaborative care models can yield better detection and depression treatment in primary care settings in which most patients with depression are seen. PMID- 16721176 TI - Personality disorders and medical comorbidity. AB - PURPOSE OF REVIEW: Personality disorders are not usually thought of as being associated with medical comorbidity. Research shows that medical comorbidity in personality disorders is clinically important. RECENT FINDINGS: In general those with personality disorders do not feel as fit as others do. Also, those with personality disorders in addition to other psychiatric disorders, such as depression and antisocial personality disorder, are likely to have more health problems than those without personality disorders. People with active borderline personality disorder have been shown to have more medical problems than those with remitted borderline personality disorder. Personality disorders can complicate the course of chronic medical illnesses. Finally, the use of psychotropic medications is not unusual in personality disorders and in itself can be associated with medical illnesses. SUMMARY: Clinicians caring for people with personality disorders need to be aware of possible medical comorbidity. More research is needed. PMID- 16721177 TI - Physical health and schizophrenia. AB - PURPOSE OF REVIEW: Patients with schizophrenia have higher than expected rates of morbidity and mortality. It is debatable whether this is related to shared risk factors, antipsychotic medication or inherent to the condition itself. This review will describe this association and the recent advances in the field. RECENT FINDINGS: The majority of patients with schizophrenia have at least one chronic comorbid medical condition. In the absence of systematic screening this may or may not be brought to the attention of health professionals. The cause of high rates of physical illness appears to be multifactorial involving shared vulnerability and genetic factors. Yet it is vascular risk factors and the adverse effects of prescribed medication that are most amenable to intervention. Current atypical antipsychotics may offer neurological and cognitive benefits, but there is accumulating evidence of problems with weight gain, diabetes, lipid dysregulation, metabolic syndrome and sexual side effects. SUMMARY: The physical health of patients with schizophrenia remains a concern. Yet the quality of medical and psychiatric care of patients with comorbid physical and mental health disorders has been shown to be unsatisfactory in several areas. We suggest that clinicians routinely assess and monitor physical health needs of patients with serious mental illness. PMID- 16721178 TI - Medical complications of anorexia nervosa and bulimia nervosa. AB - PURPOSE OF REVIEW: This review focuses on recent publications concerning medical complications in patients with eating disorders, including anorexia nervosa and bulimia nervosa. RECENT FINDINGS: Recent literature continues to reflect that multiple organ systems are frequently affected by eating disorders. The literature underscores the frequently cited risk of premature death in those with anorexia nervosa. A plethora of dermatologic changes have been described, some signaling serious underlying pathophysiology, such as purpura, which indicates a bleeding diathesis. Much of the literature continues to delineate the fact that diabetic patients with eating disorders are at high risk of developing diabetic complications. Gastrointestinal complications can be serious, including gastric dilatation and severe liver dysfunction. Acrocyanosis is common, and patients with anorexia nervosa are at risk of various arrhythmias. Low-weight patients are at high risk for osteopenia/osteoporosis. Nutritional abnormalities are also common, including sodium depletion and hypovolemia, hypophosphatemia and hypomagnesemia. Resting energy expenditure, although very low in low-weight patients, increases dramatically early in refeeding. SUMMARY: Medical complications are common and often serious in patients with eating disorders, particularly those with anorexia nervosa. PMID- 16721179 TI - Bibliography. Current world literature. Child and adolescent psychiatry. PMID- 16721180 TI - Cognitive impairment in heart failure. AB - Heart failure is reaching epidemic proportions in the United States, causing a constellation of disabling symptoms. Cognitive impairment in people with heart failure may be more common than previously known with multiple etiologies from both the disease process and the treatment. Exemplars from a research study are used to illustrate the perspective of heart failure patients. Given the critical importance of memory deficits on patient education, potential nursing interventions to address the issue of cognitive impairment are described. PMID- 16721183 TI - Abdominal pain due to acute intermittent porphyria: when is the sound of hoof beats not horses, but zebras? A case report. AB - Acute intermittent porphyria is a rare autosomal dominant hereditary inborn error of metabolism of the heme biosynthetic pathway that can be exacerbated through a multitude of environmental factors. This article is a case study describing the pathophysiology, clinical presentation management, and exacerbation prevention of acute intermittent porphyria. The disease is clinically manifested with severe abdominal pain, confusion, and seizures which may be life threatening. Specific treatment with heme preparations should be instituted as soon as increased excretion of porphobilinogen through urine sampling is confirmed. Supportive treatment includes opiate analgesia, monitoring for and treating complications such as hypertension and hyponatremia. Follow-up should include family counseling regarding genetic defects and individual counseling regarding lifestyle changes including avoidance of environmental factors that have been implicated in the exacerbation of the disease. PMID- 16721184 TI - Managing requests regarding herbal remedies for patients in the intensive care unit. AB - Nurses in intensive care units may be asked by family members to administer herbal therapies or vitamins and minerals that the patient was taking before the illness or injury. The nurse must carefully assess the request and contact the patient's healthcare providers to determine whether the request can be granted or whether the therapy poses a danger to the patient because of interactions with other medications. PMID- 16721185 TI - A diamond in the rough, to a polished gemstone ring: writing for publication in a nursing journal. AB - This manuscript will assist the novice writer with the publication process. This will walk the prospective author through selecting a topic, searching the literature, following through with the submission of the manuscript and the editorial review process, and finally emphasizing the rewards of publication. Along the way, helpful tips will be highlighted to enhance your chances of getting your manuscript accepted for publication. PMID- 16721187 TI - Strategies to meet the needs of families of critically ill patients. AB - Much research has been conducted to identify the needs of families of critically ill patients. Now is the time to develop and test various strategies to meet those needs. This article gives an overview of the identified needs of family members and potential strategies critical care nurses can use to meet these needs. PMID- 16721192 TI - Night shift: can a homeopathic remedy alleviate shift lag? AB - Night shift nurses are subject to shift lag or circadian dysrhythmia, which may result in physical and mental symptoms ranging from fatigue, irritability, depression, and apathy to gastrointestinal, cardiovascular, and sleep disorders. This study investigated the effect a homeopathic remedy No-Shift-Lag had on the night shift nurses in an intensive care unit. The study was a randomized, double blind, placebo-controlled, crossover trial. The measures included an objective computer-based vigilance test and a series of subjective questionnaires. PMID- 16721194 TI - Postmortem care. PMID- 16721197 TI - EuroPRevent Congress, Athens, Greece, 11-13 May 2006. Abstracts. PMID- 16721193 TI - Predictors of vascular complications post diagnostic cardiac catheterization and percutaneous coronary interventions. AB - Care of patients after cardiac catheterization and/or percutaneous coronary intervention is largely the responsibility of nurses. The identification of risk factors for vascular complications from these procedures is important for the development of protocols to prevent complications. This article describes a retrospective, descriptive, and correlational study of 11,119 patients who underwent cardiac catheterization and/or percutaneous intervention, with femoral artery access, in the years 2001 to 2003. Increased risk for vascular complications was found in patients who were older than 70 years, were female, had renal failure, underwent percutaneous intervention, and had a venous sheath. PMID- 16721198 TI - Usefulness and limitations of the surface electrocardiogram in the classification of right and left atrial flutter. AB - Atrial flutter is a common arrhythmia that may cause significant symptoms, including palpitations, dyspnoea, chest pain and even syncope. Frequently, it is possible to diagnose atrial flutter with a 12-lead surface electrocardiogram (ECG), looking for distinctive waves in leads II, III, aVF, aVL, V1 and V2. Puech and Waldo developed the first classification of atrial flutter in the 1970s. These authors divided the dysrhythmia into types I and II. Therefore, in 2001, the European Society of Cardiology and the North American Society of Pacing and Electrophysiology developed a new classification of atrial flutter based not only on the ECG, but also on the electrophysiological mechanism. More recently, Scheinman and colleagues have provided an updated classification and nomenclature. Terms such as common, uncommon, typical, reverse typical or atypical flutter are abandoned, because they may generate confusion. The authors worked out a new terminology, which differentiates atrial flutter only on the basis of electrophysiological mechanism. PMID- 16721199 TI - Which strategy should be used for acute ST-elevation myocardial infarction in patients aged more than 75 years? AB - The optimal management of acute myocardial infarction in elderly people (>or= 75 years) is controversial because elderly patients have been excluded or are under represented in most acute myocardial infarction trials. Randomized studies show that, also in the elderly, thrombolytic therapy is effective in reducing mortality after acute myocardial infarction but the benefit in terms of mortality, recurrent infarction and stroke is smaller compared to primary percutaneous coronary intervention. Among the available mechanical therapeutic strategies, stenting is found to be superior to balloon angioplasty, whereas the role of drug-eluting stents in this setting still remains to be evaluated. The standard use of intravenous unfractionated heparin is still recommended because of the increased risk of intracranial haemorrhage by a combination of low molecular weight heparin or IIb/IIIa inhibitors and thrombolytic agents. Dedicated randomized clinical trials are needed to establish the best reperfusion therapy for this expanding population, especially in patients admitted to hospitals without percutaneous coronary intervention facilities and in patients developing cardiogenic shock. PMID- 16721200 TI - Cardiac production of C-type natriuretic peptide in heart failure. AB - BACKGROUND: C-type natriuretic peptide production by the heart in patients with chronic heart failure has recently been demonstrated by measuring the difference in C-type natriuretic peptide plasma levels between the aortic root and coronary sinus samples. To assess this previous relevant observation, we re-evaluated the cardiac production of C-type natriuretic peptide, thus increasing the total number of patients studied. In light of the difficulty in recruiting patients with these characteristics, this kind of study can be performed with a small number of subjects. METHODS: Cardiac production of C-type natriuretic peptide was evaluated in a subset (n = 6) of patients, referred to our institute for the evaluation of chronic heart failure. C-type natriuretic peptide was measured in the coronary sinus and in aorta blood collected during right and left heart catheterization using a specific immunometric assay, after solid-phase extraction of plasma samples. RESULTS: Significantly higher C-type natriuretic peptide levels were found in the coronary sinus than in aorta blood (7.8 +/- 1.3 versus 6.1 +/- 1.5 pg/ml, P = 0.034) and our results were in accordance with previous ones. Analysing as a whole the data gathered in the two studies, a noteworthy significant increase was observed (n = 15, P = 0.002) between the C-type natriuretic peptide concentrations in the two sites. CONCLUSIONS: The results of the present study strengthen the hypothesis that C-type natriuretic peptide is produced by the heart in patients with chronic heart failure. PMID- 16721201 TI - Do mechanical markers of myocardial ischaemia predict the transmural extent of myocardial infarction in man? AB - BACKGROUND: The present study aimed to explore the relationship between the transmural extent of myocardial necrosis and mechanical markers of myocardial ischaemia in man. METHODS: A group of 40 patients with previous Q-wave myocardial infarction and a left ventricular ejection fraction (LVEF) of 27 +/- 11% was studied by cine and contrast-enhanced magnetic resonance imaging. RESULTS: Necrotic areas of delayed contrast enhancement were present in every patient and involved 20 +/- 8% of left ventricular myocardium. In involved segments, the transmural extent of contrast enhancement varied from 5% to 100%, being on average 38 +/- 25% of the wall thickness. End-diastolic left ventricular wall thickness and systolic wall thickening were lower in contrast-enhanced segments than in the other segments (P < 0.001). Furthermore, although left ventricular wall thickness and systolic wall thickening decreased as the transmural extent of contrast enhancement increased, the correlations were weak (r = -0.382 and -0.45, respectively). Finally, a delayed contrast enhancement was present in 89% of akinetic and in 94% of dyskinetic segments; however, contrast enhancement was also present in 18% of the segments with normal wall motion and in 56% of hypokinetic segments. CONCLUSIONS: Although mechanical markers of myocardial ischaemia substantially reflect the transmural extent of myocardial infarction, none of them can be considered as a substitute for the direct observation of necrotic tissue and its transmural extent, as provided by contrast-enhanced magnetic resonance imaging. PMID- 16721202 TI - Early diagnosis of congestive heart failure: clinical utility of B-type natriuretic peptide testing associated with Doppler echocardiography. AB - OBJECTIVE: B-type natriuretic peptide (BNP) has emerged as an important diagnostic serum marker of congestive heart failure (CHF). The aim of this study was to evaluate whether BNP measurement associated with echocardiography could effectively stratify patients with new symptoms as part of a cost-effective heart failure programme based on cooperation between hospital cardiologists and primary care physicians. METHODS: Patients were referred to the cardiology clinic by general practitioners in case of clinical suspect of CHF. All patients underwent clinical examination, transthoracic echocardiography and plasma determination of BNP. Systolic dysfunction was defined as a left ventricular ejection fraction < 45%; diastolic dysfunction was defined as a preserved systolic function with signs of diastolic impairment. RESULTS: Three hundred and fifty-seven subjects were examined (50% males, mean age 73 years). BNP concentration was 469 +/- 505 pg/ml in the 240 patients diagnosed with CHF, compared with 43 +/- 105 pg/ml in the 117 patients without CHF (P = 0.001). CHF patients were grouped into those with diastolic dysfunction (n = 110; BNP 373 +/- 335 pg/ml), systolic dysfunction (n = 108; BNP 550 +/- 602 pg/ml), and both systolic and diastolic dysfunction (n = 22; BNP 919 +/- 604 pg/ml). At receiver operating characteristic analysis, the optimal BNP cut-off level for diagnosing CHF was 80 pg/ml (sensitivity 84%, specificity 91%). According to cost analysis, this cut-off level might provide a cost saving of 31% without affecting diagnostic accuracy. CONCLUSIONS: In patients referred by general practitioners for suspected CHF, plasma BNP levels might help to stratify subjects into different groups of cardiac dysfunction. PMID- 16721203 TI - B-type natriuretic peptide: time to incorporate natriuretic peptides in our practice. PMID- 16721204 TI - Protective role of chronic statin therapy in reducing myocardial damage during percutaneous coronary intervention. AB - OBJECTIVE: Percutaneous coronary intervention (PCI) is frequently associated with troponin I (TnI) elevation. Experimental studies suggest that statins may reduce ischaemia-reperfusion myocardial injury. The study objective was to verify whether chronic treatment with statins might reduce the occurrence and the extent of periprocedural myocardial damage in patients undergoing PCI. METHODS: Five hundred and fifty-two consecutive patients undergoing PCI were included: 279 were not on statins before PCI, 273 were on statins. TnI levels >or= 0.3 ng/ml were considered indicative of myocardial injury. RESULTS: Statin-treated patients had a higher prevalence of hyperlipidaemia, previous myocardial infarction, and revascularization procedures. Coronary angiography also documented a higher prevalence of multivessel disease. No difference between the two groups was observed regarding the PCI-treated vessel, type of lesions, use of stents and of anti-IIb/IIIa inhibitors. Patients on statins showed the lowest incidence of TnI >or= 0.3 ng/ml (29 vs. 48%, P = 0.00001) and of creatine kinase-MB elevation (7 vs. 12%, P = 0.04). The mean peak TnI levels were significantly lower in patients on statins (1.07 +/- 3.8 vs. 2.73 +/- 12.3, P = 0.00006). Multivariate analysis identified preprocedural statin therapy as the only independent negative predictor of postprocedural abnormal TnI levels (odds ratio = 0.52; 95% confidence intervals 0.34-0.79; P = 0.003). CONCLUSIONS: This study shows that chronic therapy with statins reduces the incidence of periprocedural myocardial damage after PCI. The beneficial effect of statins was independent of either the most important clinical and angiographic characteristics or the use of other cardiovascular drugs including beta-blockers. These data support the specific cardioprotective role of statins. PMID- 16721206 TI - High take-off of the right coronary artery with intramural course associated with the persistence of the left superior vena cava. AB - We describe the case of a coronary artery with an intra-aortic course (a rare coronary anomaly) associated with the persistence of the left superior vena cava. The first finding could represent an important surgical risk factor and should be recognized before cardiac surgery; the latter is not yet reported in the literature. A modified surgical technique is described. PMID- 16721205 TI - Secondary prevention of myocardial infarction: a survey in primary care. AB - OBJECTIVE: To collect information on the major risk factors and secondary prevention among patients with myocardial infarction in Italy. METHODS: Data were obtained from the database of the Italian College of General Practitioners; 3588 patients (mean age 68.7 +/- 11.3 years; 2698 men, 888 women; two unrecorded gender), with an average time from event of 6 +/- 5.7 years, were identified. RESULTS: Among the major risk factors, data entry ranged from 50.3% for physical activity to 74.9% for blood pressure. Inadequate blood pressure control was present in 49.2% and elevated plasma cholesterol levels (> 5.2 mmol/l) in 57.3%; among the latter group, 65% were on lipid-lowering therapy. Only 47.2% of the treated patients achieved a total cholesterol level of < 5.2 mmol/l. Antiplatelet or anticoagulant drugs, beta-blockers, and angiotensin-converting enzyme inhibitors were prescribed to 43%, 10.3%, and 57.9% of patients, respectively. CONCLUSIONS: The preventive attitude of Italian general practitioners is similar to that reported in other European countries with two noticeable exceptions: under-prescription of beta-blockers and of antiplatelet drugs. Clearly, secondary prevention requires major improvement. PMID- 16721207 TI - Implantation of a biventricular pacing and defibrillator device via a persistent left superior vena cava. AB - A persistent left superior vena cava was discovered in a 66-year-old man with heart failure undergoing implantation of a biventricular pacing and defibrillator device. An active fixation right ventricular defibrillator lead was placed through a curved guiding catheter. A sub-selection catheter and a guidewire allowed the engagement of a posterior-lateral branch of the coronary sinus, performance of an angiogram without an occlusive balloon, and optimal lead placement. The right atrial lead was positioned using a standard stylet. Despite the technical challenges, implantation of a biventricular pacing and defibrillator device via a persistent left superior vena cava is safe and feasible. PMID- 16721208 TI - Electrocardiographic abnormalities mimicking myocardial infarction in a patient with intracranial haemorrhage: a possible pitfall for prehospital thrombolysis. AB - The electrocardiogram, when applied in the prehospital setting, has a significant effect on a patient with chest pain. The potential effect includes both diagnostic and therapeutic issues, including the diagnosis of acute myocardial infarction and the indication for thrombolysis or invasive procedures. We report the case of a man who suffered from a syncope, with a prehospital electrocardiogram showing prominent ST-segment elevation. Out-of-hospital thrombolytic therapy was planned by the emergency department. Fortunately, thrombolysis did not start because the patient fared worse. He was taken to the emergency department and, because of mental status impairment, it was decided to perform a cranial computed tomographic scan. The diagnosis shifted to a haemorrhagic stroke. According to the guidelines, prehospital thrombolysis would have been inappropriate in this case because the patient did not have any chest discomfort. The pathophysiological mechanisms of electrocardiographic abnormalities in the setting of intracranial haemorrhage are reviewed, as well as the issue of thrombolysis administered or planned only on the basis of an electrocardiogram. PMID- 16721209 TI - Very late thrombosis after sirolimus-eluting stent implantation. AB - Drug-eluting stents are widely used to prevent restenosis after coronary angioplasty. We here report a case of stent thrombosis within a sirolimus-eluting stent occurring two years after stent implantation, while the patient was on chronic aspirin therapy. The patient presented with acute ST-elevation myocardial infarction and pulmonary oedema; stent thrombosis was successfully treated with coronary angioplasty and deployment of a new coronary stent. This case suggests that long-term follow-up is needed in clinical studies on drug-eluting stents. PMID- 16721210 TI - Treatment of refractory heart failure with a percutaneous circulatory support device. AB - End-stage heart failure refractory to medical therapy is a clinical challenge. Heart transplantation is considered the best strategy, but it remains burdened with substantial limitations due to the scanty availability of facilities with a transplant programme, donor shortage, and unpredictable waiting lists. Alternatively, assist devices may allow an improvement of the haemodynamic conditions, modifying the history of end-stage heart failure and introducing new management strategies. The goal of a minimally invasive short-term circulatory support device in the treatment of end-stage heart failure patients awaiting heart transplant is to restore the haemodynamic conditions and to improve end organ dysfunction, with a low incidence of major adverse cardiac events. This report describes the first implantation in Italy of a new percutaneous circulatory support device, the Cancion cardiac recovery system, in a patient with severe end-stage heart failure listed for transplantation. PMID- 16721211 TI - Mechanical circulatory support is a real therapy: new perspectives in heart failure treatment. PMID- 16721212 TI - Coronary arteriovenous fistula presenting as chronic pericardial effusion. AB - We here describe the incidental finding of a rare type of anomalous coronary artery, originating from the right coronary sinus of Valsalva and draining into the right ventricle, in a 72-year-old woman admitted to our hospital with a history of chronic pericardial effusion. The presence of a coronary artery fistula was suspected on transthoracic echocardiography, but diagnosed with the use of transoesophageal echocardiography and coronary angiography. Angiography allowed delineating the origin and course of the fistula, and its drainage into the right ventricle. PMID- 16721213 TI - Left atrial appendage thrombus revealed by 16-slice cardiac computerized tomography. PMID- 16721214 TI - OESO and the Journal of Clinical Gastroenterology. PMID- 16721215 TI - GERD is chronic but not progressive. PMID- 16721216 TI - From mucosa to submucosa: a bold step forward in therapeutic endoscopy. PMID- 16721217 TI - Endoscopic submucosal dissection using flexknife. AB - Although the standard treatment for early-stage gastrointestinal tumors is still surgical resection, endoscopic resection has been accepted for some of these lesions, especially in Japan. However, the indication was limited until recently to achieve en bloc resection and prevent local recurrence. To overcome the disadvantage of endoscopic resection with conventional endoscopic mucosal resection (EMR), several investigators, including us, have developed a new endoscopic resection technique: endoscopic submucosal dissection (ESD). ESD is a remarkable technique that enables to remove the lesions en bloc regardless of size, shape, coexisting ulcer, and location. Nowadays, several knives are available for ESD, such as the needle knife, insulation-tipped (IT) knife, Hookknife, triangle-tipped (TT) knife, and Flexknife. Each of them has some merits and demerits, and the ways to use the knives are different. We summarize here how to use the Flexknife, which we made ourselves in cooperation with the Olympus Company, and how we use the technique in our hospital. PMID- 16721218 TI - Lymphogranuloma venereum in human immunodeficiency virus-infected individuals in New York City. AB - Lymphogranuloma venereum (LGV), or chlamydial proctitis, is a classic sexually transmitted disease with prominent gastrointestinal manifestations. The disease has received little attention in recent years, especially in relation to human immunodeficiency virus (HIV) infection. However, outbreaks of LGV have been reported in several large cities in Europe and the United States over the past few years, occurring in both HIV-infected and -uninfected individuals, and the reports have been largely limited to the sexually transmitted disease literature. We recently diagnosed four cases of chlamydial proctitis in HIV-infected individuals, who had different clinical presentations but very similar endoscopic and histopathologic features, as well as prompt and complete response to therapy. It is important for gastroenterologists to recognize that LGV may be reemerging as a relevant clinical entity, because of its similarity to inflammatory bowel diseases and its response to treatment with antibiotics. PMID- 16721219 TI - High-iron diet: foe or feat in ulcerative colitis and ulcerative colitis associated carcinogenesis. AB - Anemia associated with long-standing chronic inflammation and iron deficiency, and the increased risk for the development of dysplasia and carcinoma, are two of the most common complications in patients with ulcerative colitis (UC). Because of iron and nutrition deficiency, UC patients are encouraged to consume a high protein and high-iron diet. The crucial clinical question is the effect of a high iron diet on inflammation activity and inflammation-driven carcinogenesis. Is a high-iron diet a foe or a feat in UC and UC-associated carcinogenesis? This review updates the progress and information on (1) iron nutrition and iron deficiency anemia in patients with UC, (2) experimental evidence of the exacerbating effect of a high-iron diet on UC and its associated carcinogenesis and the difference between a high-iron diet and parental iron supplementation, (3) the clinical efficacy of, and concerns about, oral and intravenous iron supplements in patients with inflammatory bowel disease and iron deficiency anemia, and (4) the clinical implications of long-term iron supplements and management of UC. These experimental findings from animal models provide evidence to warrant further consideration and clinical studies of iron nutrition, inflammation activity, and cancer development. PMID- 16721220 TI - The long-term natural history of gastroesophageal reflux disease. AB - INTRODUCTION: Long-term gastric acid suppression has been suggested as a means to prevent complications of reflux esophagitis. We report on the 20-year follow-up of 2,306 patients with at least two endoscopic examinations who were taking no antisecretory medication before baseline endoscopy and whose long-term treatment was determined by reflux symptoms. METHODS: From 1979 through 1998, endoscopy and biopsy were performed in the Hines Veterans Affairs Hospital endoscopy clinic by three endoscopists. Antireflux treatment was symptom-driven, and endoscopies were repeated mostly for symptomatic recurrence due to cessation of therapy. RESULTS: Of 4,633 patients undergoing endoscopy for reflux symptoms, 2,306 had at least one follow-up endoscopy and biopsy. Over a mean follow-up period of 7.6 years (range, 1-20 years), the esophageal mucosa of 67% of patients remained unchanged, that of 21% improved, and that of 11% worsened. Esophageal stricture requiring dilation developed from a normal baseline mucosa in one of 1,313 patients (0.08%) and from an erosive baseline mucosa in 18 of 957 patients (1.9%). The overall incidence of stricture in patients with gastroesophageal reflux (GER) disease was <1/1,000 per year. Nonsteroidal anti-inflammatory drug (NSAID) consumption was associated with less mucosal improvement (odds ration [OR] = 0.67; confidence interval [CI] = 0.46-0.98). Use of histamine-2 receptor antagonists (H2RAs) and proton pump inhibitors (PPIs) was associated with mucosal improvement (OR for PPIs = 1.49; CI = 1.14-2.17). Cohn's kappa was 42%, confirming the results that demonstrate stability of esophageal mucosal disease in the majority of patients. CONCLUSIONS: Symptom-driven treatment of GER disease after a thorough endoscopic examination to exclude premalignant or malignant esophageal mucosal disease is practical and safe for the vast majority of patients with uncomplicated GER symptoms. PMID- 16721221 TI - Association of esophageal adenocarcinoma with other subsequent primary cancers. AB - PURPOSE: Esophageal adenocarcinoma shares risk factors such as obesity and smoking with other common cancers. The association of esophageal adenocarcinoma with other primary cancers has not been systematically evaluated. The authors used the Surveillance, Epidemiology and End-Results database of the National Cancer Institute to explore the association of esophageal adenocarcinoma with other primary cancers. PATIENTS AND METHODS: All adult patients with esophageal cancers, both adenocarcinoma and squamous cell carcinoma diagnosed between 1973 and 2001, were identified from the Surveillance, Epidemiology, and End-Results database, and standardized incidence rates were calculated for all subsequent primary cancers in these patients. The analysis was reversed to estimate the standardized incidence rate for subsequent primary esophageal cancer after a first primary cancer. RESULTS: In comparison with a standard population, patients with esophageal adenocarcinoma were at higher risk for the development of another subsequent cancer, specifically, cancers of the oral cavity and pharynx, lung and bronchus, and kidney and renal pelvis, and adenocarcinoma of the colon/rectum and pancreas. With squamous cell esophageal cancer, there was an association with tobacco-related cancers such as those of the oral cavity and pharynx, the lung and bronchus, and the breast. There was either no association or even a negative association of esophageal adenocarcinoma with other obesity-related cancers such as breast, uterine, and prostate cancers. CONCLUSIONS: Patients with esophageal adenocarcinoma and squamous cell carcinoma are at increased risk for the development of specific second primary cancers that share smoking as a common risk factor. Esophageal adenocarcinoma does not have a strong association with obesity-related cancers with respect to the relative risk for the development of subsequent primary cancers. PMID- 16721223 TI - Dyspepsia in homeless adults. AB - GOALS: To determine the prevalence of and risk factors for dyspepsia in a representative sample of homeless persons using shelters in Toronto, Canada. BACKGROUND: Homeless people have many risk factors for dyspepsia, but little information is available on gastrointestinal symptoms in this population. STUDY: Cross-sectional survey of a representative sample of 100 homeless adults, with serologic testing for Helicobacter pylori infection. RESULTS: The prevalence of moderate, severe, or very severe symptoms within the past 3 months was 18% for upper stomach pain and 59% for any dyspeptic symptom. Nonwhite ethnicity (odds ratio, 3.5; 95% confidence interval, 1.1-10.9) and a history of gastrointestinal disease (odds ratio, 8.6; 95% confidence interval, 2.5-29.6) were significantly associated with moderate to very severe upper stomach pain. H. pylori infection was identified in 31% of participants but was not significantly associated with dyspepsia. CONCLUSIONS: Dyspepsia is a common problem among homeless adults in Toronto. The presence of upper stomach pain is most strongly associated with a history of gastrointestinal disease. PMID- 16721222 TI - Predictive factors of lymph node metastasis in patients with undifferentiated early gastric cancers. AB - BACKGROUND: For intramucosal differentiated early gastric cancer that has little risk of lymph node metastasis, local treatment such as endoscopic mucosal resection has been generally accepted as an adequate treatment. We studied clinicopathological characteristics of undifferentiated early gastric cancer at our institution to identify the predictive factors for lymph node metastasis and qualify lesions that should be referred for gastrectomy and not endoscopic mucosal resection. METHODS: We retrospectively analyzed the clinicopathological features (patient age and gender, tumor size, location, macroscopic type and histological type, presence of ulceration, depth of tumor invasion, and lymphatic vascular involvement) in 332 patients with undifferentiated early gastric cancer who underwent gastrectomy with regional lymph node dissection. RESULTS: Lymph node metastasis was observed in 45 patients (14%). Univariate analysis revealed that depth of tumor invasion (submucosa), tumor size (>30 mm), and lymphatic vascular involvement (positive) were associated with lymph node metastasis. Only lymphatic-vascular involvement (positive) was found to have a significant association (odds ratio, 7.4; 95% confidence interval, 2.9-19.0) by multivariate analysis. CONCLUSIONS: Lymphatic-vascular involvement was the only independent predictive risk factor for lymph node metastasis. This pathologic factor was not useful for identifying patients at high risk of lymph node metastasis who should be offered gastrectomy rather than endoscopic mucosal resection. PMID- 16721224 TI - Volatile sulfur compounds produced by Helicobacter pylori. AB - GOALS: To assess the volatile sulfur compounds produced by three strains of Helicobacter pylori in broth cultures mixed with sulfur-containing amino acids. BACKGROUND: Halitosis has been reported in H. pylori-positive patients, and volatile sulfur compounds such as hydrogen sulfide and methyl mercaptan are known to be responsible for inducing oral malodor. Whether H. pylori produces these volatile sulfur compounds has yet to be established. STUDY: Three strains of H. pylori (ATCC 43504, SS 1, DSM 4867) were cultured with 5 mM cysteine and methionine. After 72 hours of incubation, the headspace air was aspirated and injected directly into a gas chromatograph. The concentrations of hydrogen sulfide and methyl mercaptan were analyzed and compared between experimental and control cultures RESULTS: In broth containing 5 mM cysteine, hydrogen sulfide was increased by ATCC 43504 (P < 0.01) and SS 1 (P < 0.05), while methyl mercaptan was elevated only by SS 1 (P < 0.05). In broth containing 5 mM methionine, methyl mercaptan increases were significant for SS 1 (P < 0.05) and DSM 4867 (P < 0.05). In broth containing 5 mM cysteine and 5 mM methionine, the concentration of hydrogen sulfide was higher than in controls for all three strains (P < 0.01); that of methyl mercaptan was higher only for SS 1 (P < 0.01). Cysteine addition to cultures containing methionine increased hydrogen sulfide and methyl mercaptan for ATCC 43504 (P < 0.05) and SS 1 (P < 0.05). Conversely, addition of methionine to cultures containing cysteine increased methyl mercaptan only for DSM 4867 (P < 0.01). CONCLUSIONS: The production of volatile sulfur compounds by H. pylori is not only very complicated but also strain-specific. Nevertheless, H. pylori was shown to produce hydrogen sulfide and methyl mercaptan, which suggests that this microorganism can contribute to the development of halitosis. PMID- 16721225 TI - Peritoneal mice implicated in intestinal obstruction: report of a case and review of the literature. AB - BACKGROUND: Peritoneal loose bodies or "peritoneal mice" are asymptomatic and mostly found as "incidentalomas" during intraperitoneal surgery or at autopsy. Implication of these concretions in an acute abdomen is a rare clinical entity. CASE REPORT: We report the case of a 63-year-old man who presented with small bowel obstruction and two calcified concretions in his pelvis. A computerized tomography scan demonstrated two oval stones near the terminal ileal loop with lumenal compression of the distal small bowel. During laparotomy, two extralumenal glistening stones, measuring 5.8 x 4.5 x 3.7 cm and 5.2 x 4.5 x 3.7 cm, were recovered from the peritoneal cavity. Histologically, the wall showed concentric lamellar fibrosis and calcification, whereas the nucleus displayed the vestiges of membranous fat necrosis; all consistent with diagnosis of peritoneal loose body. All symptoms of bowel obstruction were relieved postoperatively, and the patient had an uneventful recovery. DISCUSSION: Peritoneal loose body is a histopathologic diagnosis, and most other differentials for calcified intraperitoneal masses can be ruled out during the histologic examination. Surgical removal is recommended because conventional cross-sectional imaging cannot accurately establish the diagnosis. PMID- 16721227 TI - A search for acute necrotic pancreatitis in early stages of alcoholic chronic pancreatitis. AB - OBJECTIVES: Various investigators believe that alcoholic chronic pancreatitis is the result of recurrent episodes of acute necrotic pancreatitis. The aim of this work is to study pancreatic histology in early stages of the disease to search for evidence of these acute episodes. STUDY: Of about 650 patients with alcoholic pancreatitis seen during the 30-year period from 1972 to 2002, 45 underwent surgery for this disease, all within 2 years of clinical onset. Of these 45, tissue samples adequate for study were obtained from 42, and this was the study material. Tissue samples were prepared for histologic examination by standard procedures. RESULTS: Areas of pancreatic necrosis were seen in tissue samples of only three (7%) of the 42 patients, and in all three cases chronic lesions were also present. No evidence of localized scarring that could be attributed to prior episodes of focal necrotic pancreatitis was found. A typical feature was the patchy distribution of the lesions in largely normal pancreatic tissue. The main lesions observed were perilobular and intralobular fibrosis, dilation of acini and ducts, and protein plugs in dilated ducts surrounded by periductal fibrosis. CONCLUSIONS: This study shows that, in the early stages of alcoholic chronic pancreatitis, signs of acute necrotic pancreatitis are very infrequent and, when present, they are associated with chronic lesions. These findings suggest that alcoholic pancreatitis begins as a chronic disease. PMID- 16721226 TI - Early enteral nutrition in severe acute pancreatitis: a prospective randomized controlled trial comparing nasojejunal and nasogastric routes. AB - PURPOSE: Enteral nutrition (EN) is effective, easy to provide, cheaper, and associated with fewer complications in comparison with parenteral nutrition in severe acute pancreatitis (SAP). However, the nasogastric (NG) route for enteral supplements still remains to be established, and most studies have used the nasojejunal (NJ) route. The purpose of this study was to compare early NJ with NG feeding in SAP. PATIENTS AND METHODS: A total of 31 patients with SAP were randomized to feeding by either NG (15 patients) or NJ (16 patients). A semi elemental formula was used through an enteral tube in both groups. Nutritional parameters (anthropometry, serum prealbumin and albumin levels) were recorded at baseline and after 7 days. Recurrence of pain and tolerance of feeding was noted. RESULTS: Recurrence of pain occurred in only 1 patient each in the 2 groups. Diarrhea occurred in 3 and 4 patients in the NJ and NG groups, respectively. There were 4 deaths in the NJ group and 5 in the NG group. Two patients in the NJ group and 1 in the NG group underwent surgery. There was no difference in the outcome measures (ie, discharge, surgery, and death). There was a decline in nutritional parameters in both groups. CONCLUSIONS: EN at a slow infusion is well tolerated by both NJ and NG routes in patients with SAP. Neither NJ nor NG feeding leads to recurrence or worsening of pain in SAP. Nutritional parameters remained unaffected because of inadequate calorie intake during the first week of feeding. PMID- 16721229 TI - Is hepatitis C more aggressive in renal transplant patients than in patients with end-stage renal disease? AB - BACKGROUND: The eventual impact of immunosuppression on the natural history of hepatitis C virus (HCV) infection in patients with end-stage renal disease (ESRD) is still unknown because of the lack of comparative data for HCV-infected patients with ESRD and renal transplant patients. The aim of this study was to compare the biochemical and histological characteristics of chronic HCV infection in renal transplants patients and ESRD patients undergoing hemodialysis. METHODS: Thirty-eight renal transplant patients and 38 ESRD patients undergoing hemodialysis who were chronically infected with HCV and were matched for gender, age at infection, and estimated time of infection were included in the study. The groups were compared regarding laboratory and histological variables. RESULTS: Renal transplant patients showed similar alanine aminotransferase and higher gamma-glutamyltransferase levels (P = 0.05) when compared with ESRD patients. Comparative analysis of histological variables revealed a higher proportion of cases with septal fibrosis (P = 0.04) and confluent necrosis (P = 0.01) among transplant-recipient patients. No difference between groups was observed regarding the intensity of portal and periportal inflammatory infiltrates. Steatosis was more prevalent among transplant-recipient patients (P < 0.001). There was no difference between groups regarding the prevalence of lymphoid aggregates or bile duct injury. CONCLUSION: Renal transplant patients had a larger proportion of cases with septal fibrosis and confluent necrosis than did ESRD patients, suggesting that renal transplantation might modify the natural history of hepatitis C in ESRD patients, leading to a more aggressive liver disease. PMID- 16721228 TI - Lack of higher frequency of the chemokine receptor 5-delta32/delta32 genotype in hepatitis C. AB - PURPOSE: An elevated frequency of the CCR5-Delta32 mutation in German patients with hepatitis C with viremia has been reported. The aim of the present study was to verify whether this mutation occurs in an Italian population with hepatitis C and whether it is an adverse host factor indicative of severity of liver disease and response to antiviral therapy. STUDY: The authors amplified 189-bp (wild type) and 157-bp (Delta32 deletion) fragments of the CCR5 gene by polymerase chain reaction in 130 patients with chronic hepatitis C. Comparisons were drawn with 110 blood donors and 135 patients with primary biliary cirrhosis. RESULTS: Four (3.1%) patients with chronic hepatitis C and 1 blood donor (0.9%) were CCR5 Delta32 homozygous, whereas there was no CCR5-Delta32 homozygosity among primary biliary cirrhosis patients; the wild-type gene was present in a similar percentage in the 3 groups of patients without any significant difference (83.1% vs 90.4% vs 83.6%, respectively). Among the patients with chronic hepatitis C, no significant correlation was found between CCR5-Delta32 homozygosity and the following parameters: histologic grade/stage, hepatitis C virus genotype, viral load, serum aspartate aminotransferase, serum alanine aminotransferase, and serum gamma-glutamyltransferase. Ninety-five patients received a standard antiviral protocol with pegylated interferon (PEG Intron)+ribavirin; a sustained response was achieved in 59 patients (62.1%), and the remainder did not respond or experienced a relapse. Response to treatment was not influenced by CCR5-Delta32 deletion. CONCLUSION: No greater frequency of CCR5-Delta32 homozygosity was seen in an Italian population of patients with chronic hepatitis C. This mutation does not seem to influence either the overall severity of liver disease or the response to viral therapy. PMID- 16721230 TI - "Endoscopy blues". PMID- 16721231 TI - Treatment of type II cryoglobulinemia associated with hepatitis C with rituximab. PMID- 16721232 TI - Constrictive pericarditis in a patient with Turner's syndrome and liver cirrhosis: coincidence or part of the syndrome? PMID- 16721233 TI - Multiple biliary stents: a therapeutic option for persistent biliary leak following laparoscopic cholecystectomy. PMID- 16721234 TI - "Medical treatment of portal biliopathy". PMID- 16721235 TI - Streptococcus bovis bacteremia associated with acute cholecystitis. PMID- 16721236 TI - Enterobacter cancerogenus bacteremia in a patient with poor dentition, cirrhosis, and a variceal bleed. PMID- 16721237 TI - Invasive and noninvasive investigations for pediatric nonalcoholic liver fatty disease: time to move beyond liver biopsy. PMID- 16721239 TI - Reconstruction of distal tibia fractures using a posterolateral approach and a blade plate. AB - OBJECTIVE: The aim of this article is to report a technique for the management of distal tibia fractures with significant anteromedial soft-tissue injury. The patients were initially treated with a spanning external fixator, open reduction and internal fixation (ORIF) of the fibula at the discretion of the surgeon, and soft-tissue management or flap coverage. ORIF of the tibia was performed on a staged basis, using a 90-degree cannulated blade plate and autogenous iliac crest bone graft through a posterolateral approach. DESIGN: Retrospective analysis of a consecutive series of patients. SETTING: Two academic level-1 trauma centers. PATIENTS: Fifteen patients with 15 distal tibia fractures (13 open fractures), Orthopedic Trauma Association (OTA) type 43A3 and 43C1, were definitively treated and followed to union between July 2000 and July 2004. Five patients were referred from outside sources after initial stabilization. INTERVENTION: Initial stabilization in an external fixator and management of the open fracture and soft tissue. Staged ORIF of the tibia with bone graft was performed through a posterolateral approach when the soft tissues allowed. OUTCOME MEASUREMENTS: Radiographic union, American Orthopaedic Foot and Ankle Society (AOFAS) ankle hindfoot score, and complications. RESULTS: All 15 fractures were followed to union. Average time to union was 20 (12 to 47) weeks from the time of fixation with blade plate and bone grafting. (AOFAS) ankle-hindfoot score was used to measure outcome. The average score was 81 (60 to 97) out of a possible 100. There were no deep infections. There was one nonunion; the fracture united after revision with a locked plate and bone graft. The average length of follow-up was 14 months (4 to 37). CONCLUSIONS: The staged treatment of high-energy distal tibia fractures with soft-tissue injury can lead to good outcomes and consistent bone union. Our results were obtained by the combination of the posterolateral approach, careful soft-tissue management, and stable internal fixation. PMID- 16721240 TI - Unexpected high complication rate following internal fixation of unstable proximal humerus fractures with an angled blade plate. AB - OBJECTIVES: The increasing number of fractures of the proximal humerus, especially in the elderly, carries with it the unsolved problem of the optimal treatment for the displaced or unstable fractures. The authors' goal was to analyze whether fixation with a bent valgus angled blade plate could improve the outcomes reported in the literature to date. DESIGN: Prospective clinical study. SETTING: Urban level 1 university trauma center. PATIENTS: Over a 27-month period, 42 consecutive patients were treated for an unstable or displaced proximal humerus fracture. INTERVENTION: Open reduction and internal fixation with a 90-degree cannulated angled blade plate prebent to 110 degrees. MAIN OUTCOME MEASUREMENTS: Active follow-up for 1 year with assessment of objective and subjective functional results (ie, motion; strength; Constant score; Disabilities of the Arm, Shoulder, and Hand (DASH) score; and visual analog scale (VAS)) and radiographic assessment (reduction, alignment, necrosis, and nonunion). RESULTS: Follow-up was completed for 86% of the patients, who achieved a mean Constant score of 66 points (82% of the contralateral side) with a mean forward flexion of 125 degrees and an average strength of 72% of the contralateral side. Low disability (mean DASH score 22) and pain values (mean VAS 2) were demonstrated after this type of stabilization. The overall complication rate was 33% (12/36), with protrusion of the blade into the glenohumeral articulation as the most frequent problem (8/36, 22%). The negative impact of an adverse event on subjective and objective outcomes was only significant for forward flexion (P = 0.02). Neither clinical outcome nor complication rate was different when compared to patients with regard to fracture type (3 versus 4 parts) or age (younger or older than 70 years). CONCLUSIONS: Fixation of displaced proximal humeral fractures with an angled blade plate provided sufficient stability. Blade perforation into the humeral joint occurred in every fourth patient and was found to be the major reason for a high complication rate. In view of this major problem, the technique described here cannot be recommended, even though the absence of nonunions in our series seems to support the low invasiveness of this surgical approach. PMID- 16721238 TI - Cephalomedullary nails in the treatment of high-energy proximal femur fractures in young patients: a prospective, randomized comparison of trochanteric versus piriformis fossa entry portal. AB - OBJECTIVE: The purpose of this study is to compare a cephalomedullary nail that uses a piriformis fossa starting point to one that uses a trochanteric starting point, in the treatment of high-energy proximal femur fractures in young patients. Our hypothesis was that a nail that uses a trochanteric starting point would result in less blood loss than a nail that uses a piriformis fossa starting point. DESIGN: Prospective, randomized. SETTING: Level 1 trauma center. PATIENTS: Thirty-four consecutive patients aged between 18 and 50 years who sustained a subtrochanteric, intertrochanteric, or ipsilateral femoral neck/shaft fracture due to a high-energy injury were enrolled. INTERVENTION: Patients were randomized to have their fractures repaired with a Russell-Taylor Recon Nail or Howmedica Long Gamma Nail. Surgery was performed on a fracture table, in supine or lateral position according to the surgeon's preference. Direct fracture exposure was avoided. Reduction was obtained through traction, patient positioning, and manual pressure. If necessary, stab-wound incisions were made to introduce instruments to improve reduction. Intramedullary reamers were used, and all nails were statically locked. Bone grafting was not used. MAIN OUTCOME MEASURES: Blood loss, incision length, duration of surgery, and body mass index were recorded for each patient. Surgeon's assessment of ease of use of the device and quality of reduction were noted. Patients were to be followed up to assess fracture union. Hip and knee ranges of motion at latest follow-up were measured. Radiographs obtained at the time of union were assessed for varus malalignment. Return to work status was recorded, and the Harris Hip Score was used to assess hip function. RESULTS: There were 17 patients in each group. The 2 groups did not differ with regard to blood loss, incision length, and duration of surgery or intraoperative complications. Body mass index was significantly linked to duration of surgery (P<0.001) and incision length (P<0.001). Surgeon's assessment of ease of use and reduction quality for the two devices did not differ. The rate of varus malunion did not differ between the 2 groups. Two patients were lost to follow-up before fracture union. All other fractures healed with no need for bone grafting or other procedures to obtain union. One obese patient developed a wound infection that resolved after debridement and a course of antibiotics. A total of 6 patients were lost prior to their 1-year follow-up visit. Among the remaining 28 patients, at an average follow-up of 14 months, no difference was noted between the 2 groups with regard to return to work status, Harris Hip Score, or hip and knee ranges of motion. CONCLUSIONS: Both devices yield predictably good results in these difficult fractures. We found no difference between the two devices with regard to incision length, duration of surgery, blood loss, reduction, ease of use, union rate, complication rate, or outcome. PMID- 16721242 TI - The ability of the Lauge-Hansen classification to predict ligament injury and mechanism in ankle fractures: an MRI study. AB - OBJECTIVES: The Lauge-Hansen classification system was designed to predict the mechanism and ligament injury patterns of ankle fractures on the basis of x-rays. The purpose of this study was to evaluate the accuracy of these predicted injury sequences using magnetic resonance imaging (MRI) in a series of patients with ankle fractures. DESIGN: Retrospective cohort. SETTING: Two university level 1 trauma centers. PATIENTS: Fifty-nine patients with operative ankle fractures who were evaluated with both x-ray and MRI were included. INTERVENTION: All patients had a standard 3-view ankle x-ray series before fracture reduction, followed by an MRI. All plain x-rays were assigned to a Lauge-Hansen category by an experienced orthopedic traumatologist. MRI studies were subsequently read by an MRI musculoskeletal radiologist for the integrity of the ankle ligaments. MAIN OUTCOME MEASUREMENTS: After evaluation of the x-rays, fractures were classified according to the system of Lauge-Hansen, and the predicted presence, sequence, and mechanism of injury was determined. These were then compared to the actual injured structures on MRI in each case, and the ability of the Lauge-Hansen system to accurately predict the complete injury pattern was determined for the entire cohort. RESULTS: Average patient age was 59 (range: 18 to 84) years. Of the 59 ankle fractures evaluated, 37 (63%) were classified as supination external rotation, 11 (19%) were pronation external rotation, 1 (2%) was supination adduction, and 10 (17%) were not classifiable on the basis of the Lauge-Hansen system. Of the 49 fractures that fit into Lauge-Hansen categories, 26 (53%) had patterns of ligamentous injury and fracture morphology that did not coincide with the Lauge-Hansen predictions. A common fracture pattern was observed in 8 of the 10 unclassifiable fractures, which included a high spiral fracture of the fibula, vertical shear fracture of the medial malleolus, posterior malleolar fracture, and complete tears of the anterior-inferior tibiofibular ligament and the interosseous membrane. In addition, over 65% of patients in this series had complete ligamentous injury and a fracture of the malleolus to which the ligament attaches. CONCLUSIONS: These results demonstrate that the Lauge-Hansen classification system may have some limitations as a predictor of the mechanism of injury and the presence of soft-tissue damage associated with ankle fractures. The identification of a novel pattern of ankle fracture also illustrates how the system fails to describe all possible fracture patterns. For these reasons, we recommend that the Lauge-Hansen system be used only as a guide in the diagnosis and management of ankle fractures and not solely relied upon for treatment decisions. Although the exact clinical implications of the variety of ligamentous injuries observed on MRI are yet to be determined, this technique may be useful in individual cases in which doubt about joint stability and soft-tissue integrity exists. Additionally, MRI may be helpful in planning surgical approaches in atypical fractures in which injury patterns are less predictable solely on the basis of x-ray. PMID- 16721241 TI - Effects of varying pulsatile lavage pressure on cancellous bone structure and fracture healing. AB - OBJECTIVE: To study the effects of variations in pulsatile lavage irrigation pressure on the rate of new bone formation and the degree to which cellular elements are removed from cancellous bone after fracture. DESIGN: A previously described intraarticular fracture model was used for 29 New Zealand white rabbits that underwent osteotomy of the medial femoral condyle. Fractures were irrigated with high-pressure pulsatile lavage at a fixed distance and volume, but at nozzle pressures varying from 20 to 70 psi. Fractures were reduced and stabilized, and animals euthanized 14 days after fracture. Fluorescent bone staining was used to determine the rate of new bone formation in the osteotomy site. At the time of euthanasia, the nonoperated knees of 12 rabbits underwent osteotomy and irrigation using the same protocol. These specimens were sent for immediate scanning electron microscopy to determine the amount of cellular material removed from the bony trabeculae. RESULTS: In the first week after irrigation, there was no significant difference in the amount of new bone formation between the 20- and 30-psi groups, but there were significant differences between these groups and the 50- and 70-psi groups. There were no significant differences between any of the groups in the amount of new bone formed during the second week after irrigation. No structural damage to the bony trabeculae was observed in any specimen irrigated at any of the pressures used. There was a direct correlation between percentage of the trabecula completely cleared of cellular material and irrigation pressure; there were statistically significant differences between each of the groups. CONCLUSIONS: There are presently no recommended guidelines as to the optimal irrigation pressure, and this study is the first to address the effects of variations in pressure on bone healing. The results of this study indicate that early new bone formation in an intraarticular fracture rabbit model is inhibited by irrigation pressure of 50 psi or greater. Additionally, this study demonstrates a direct relationship between irrigation pressure and the amount of cellular material removed from the trabecula at the irrigation site. Surgeons should be aware of the potentially detrimental effects of using irrigation pressures at or above 50 psi in the treatment of fractures. PMID- 16721243 TI - A mechanical evaluation of two fixation methods using cancellous screws for coronal fractures of the lateral condyle of the distal femur (OTA type 33B). AB - PURPOSE: To compare the relative strength and stability of 2 fixation methods for displaced coronal shear fractures of the lateral femoral condyle (Hoffa fractures, OTA Type 33B3). SETTING: University Biomechanics laboratory. DESIGN: Eight matched pairs of embalmed femurs were divided into 2 groups and simulated Hoffa fractures were created. In each pair, 1 of the fractures was fixed with 2 screws placed in an anteroposterior direction, and in the other, the fracture was fixed with 2 screws placed in a posteroanterior direction. METHODS: All specimens were cyclically tested with simulated physiologic loading. Displacement of the femoral condyle was continuously measured to 10 cycles. The specimens were then loaded to failure. RESULTS: Fixation with posterior to anteriorly placed cancellous lag screws was significantly more stable than that with anterior to posteriorly placed screws at 10 cycles (P = 0.05), with 0.67 mm displacement compared to 1.36 mm, respectively. They were also more stable at 10, 100, and 1000 cycles; however, these displacements were not statistically significant. Fixation with posteriorly placed cancellous screws also had significantly higher ultimate strength (P = 0.04), 1700 N compared to 1025 N for anterior placement. CONCLUSION AND SIGNIFICANCE: Lag screws placed posterior to anterior provided more stable fixation of Hoffa fractures in embalmed femurs than anteroposteriorly placed lag screws. This finding may apply in the clinical setting; however, this technique requires that the screw heads be recessed beneath the articular surface. The effects of the cartilage defects so created are not known. The choice of technique is also determined by concomitant fractures and the exposure required for their fixation. PMID- 16721246 TI - Semiconstrained knee arthroplasty in the setting of a chronic knee dislocation: a case report. AB - We present the case of a 27-month chronic knee dislocation treated with a semiconstrained, stemmed total knee arthroplasty. The patient was neurovascularly intact preoperatively but had severe functional limitations caused by pain, stiffness, and instability. At 2-year follow-up, the patient remains pain-free with functional range of motion and the ability to ambulate without complication. This case report represents the longest-cited chronic knee dislocation treated with a reconstructive procedure. PMID- 16721252 TI - Treatment options in pediatric femoral shaft fractures. PMID- 16721244 TI - Reduction and pinning of pediatric supracondylar humerus fractures in the prone position. AB - Supracondylar humerus fractures are common pediatric injuries; the preferred treatment for displaced fractures is closed reduction and percutaneous pin fixation. We present a technique for closed reduction and pinning using prone patient positioning. Prone positioning facilitates fracture reduction and safe pin placement while avoiding elbow hyperflexion. We prefer this technique to the commonly described method of fracture reduction and pinning with the patient supine. PMID- 16721248 TI - Isolated intra-articular fracture of the distal ulna: a rare injury related to airbag deployment. AB - An isolated intra-articular and adjacent shaft fracture of the distal ulna is described. A 38-old-year woman driver sustained an automobile accident with subsequent airbag deployment injuring her left wrist. Imaging studies showed displacement of a radiovolar fragment and deformity of the ulnar articular surface. The fracture was reduced and internally fixed with 3, 3.5-mm, cannulated screws to avoid future difficulties in forearm rotation, ulnocarpal impaction, and late posttraumatic arthritis. The final outcome was excellent. Careful clinical and radiologic examination is strongly recommended for this type of fracture, which seems to be a part of the spectrum of upper limb injuries associated with airbag deployment. Such injuries can be expected to occur more frequently with the increasing use of airbags. PMID- 16721250 TI - Bending and torsional stiffness in cadaver humeri fixed with a self-locking expandable or interlocking nail system: a mechanical study. PMID- 16721254 TI - Penetration of the distal femoral anterior cortex during intramedullary nailing for subtrochanteric fractures. PMID- 16721247 TI - Traumatic abdominal hernia and lateral compression type 1 pelvic fracture: a case report. AB - Pelvic fractures often are associated with concomitant injuries. In general, the more severe the pelvic fracture, the more likely other, potentially life threatening injuries exist. We present a case of a typical type 1 lateral compression pelvic fracture with the less common associated injury of abdominal wall muscle disruption and large-bowel herniation. PMID- 16721245 TI - The anconeus flap transolecranon approach to the distal humerus. AB - The transolecranon approach for the treatment of distal humerus fractures and nonunions is commonly used. A complication of the standard osteotomy is denervation of the anconeus muscle, which provides dynamic stability to the lateral side of the elbow by preventing varus and posterolateral rotatory instability. This article describes the anconeus flap transolecranon (AFT) approach, which utilizes an internervous plane to preserve the anconeus muscle and a chevron-shaped osteotomy for maximal joint exposure. The approach is straightforward to perform with limited complications. PMID- 16721255 TI - Pernicious anemia in Chinese: a study of 181 patients in a Hong Kong hospital. AB - To study the clinical and hematologic features of pernicious anemia in Chinese, we describe 181 Chinese with megaloblastic anemia and low serum cobalamin, in association with either classic Schilling test results (82 patients) or the presence of serum antibody to intrinsic factor (99 patients), encountered in a regional hospital in Hong Kong from May 1994 to May 2005. The median age was 75 years (range, 32-95 yr) and the male to female ratio was 1:1.5. The chief presenting feature was anemia, and fewer than 10% of patients presented predominantly with neurologic deficit. Gastric biopsies of 109 patients showed glandular atrophy in 73, endocrine cell hyperplasia in 5, polyps in 14, adenocarcinoma in 1, and chronic gastritis in the rest. Gastric adenocarcinoma occurred in 1.7% of patients after a median follow-up of 35 months (range, 0.5 132 mo). Diabetes mellitus occurred in 24% of patients and thyroid disease in 7%. No specific ABO blood group was associated with pernicious anemia. Serum antibody to intrinsic factor (73%) occurred more frequently than serum antibody to gastric parietal cell (65%) (p=0.353). The frequency of serum antibody to gastric parietal cell was higher in male (78%) than in female patients (53%) (p=0.018). Pernicious anemia is a major cause of megaloblastic anemia in Chinese. PMID- 16721258 TI - Systemic lupus erythematosus in Asturias, Spain: clinical and serologic features. AB - Asturias is an autonomous region in the north of Spain with historical and anthropologic peculiarities. In the current report, we examine the main clinical and immunologic features of 363 patients with systemic lupus erythematosus (SLE), virtually the entire population of SLE patients in Asturias. We constructed a database with the clinical and immunologic features of all patients fulfilling the American College of Rheumatology criteria, based on the review of hospital records corresponding to blood samples received for antinuclear antibodies testing since 1992. Arthritis was the most frequently observed main clinical feature and neuropathy was the rarest. Male patients had a disease more frequently characterized by serositis (p<0.05) and neurologic disorder (p<0.01) than females, while children presented malar rash (p<0.05), fever (p<0.05), and kidney involvement (p<0.01) more often than adults. Late-onset patients were characterized by lower frequencies of malar rash (p<0.01), neurologic disorder (p<0.05), alopecia (p<0.01), and lymphadenopathy (p<0.05) than young adults. Numerous direct and inverse significant associations were found among clinical and immunologic features. The most relevant significant associations were neurologic disorder with lupus anticoagulant (p<0.01); kidney involvement with serositis (p<0.01) and DNA antibodies (p<0.05); and thrombosis with DNA antibodies (p<0.05), cardiolipin antibodies (p<0.01), and lupus anticoagulant (p<0.01). A low mortality was found in our series, although kidney involvement (p<0.05) and cardiolipin antibodies (p<0.05) are factors associated with poor survival. PMID- 16721260 TI - Hypertrophic osteoarthropathy secondary to vascular prosthesis infection: report of 3 cases and review of the literature. AB - An infected vascular graft is an uncommon association in hypertrophic osteoarthropathy (HOA) but is important because of potential catastrophic complications. Here we describe 3 new patients with HOA secondary to vascular prosthesis infection and review the other 23 patients described in the English literature with this clinical syndrome. The finding of symptoms and signs of localized HOA in patients with arterial prosthesis may be the presenting manifestation of graft sepsis. In most patients the graft is located at the aorta or aortic bifurcation. The initial symptoms are fever, joint pain mainly involving the knees and ankles, clubbing, and in some cases intestinal bleeding. Recurrent abscesses and bacteremia arising in spite of appropriate antimicrobial therapy are prominent infectious signs. Another characteristic feature is the wide variety of bacteria isolated from a single patient. When a polymicrobial enteric flora is isolated from the blood or other tissue, an intestinal origin of infection can be readily suspected. Gallium or leukocyte scans are useful for detecting an abscess around the vascular prosthesis. In some cases, the detection of gas around the graft demonstrated by computed tomography is highly suggestive of prosthesis infection. Mechanisms involved in the pathogenesis of HOA associated with aortic graft infection are unknown. Treatment consists of intravenous antibiotics and surgical replacement of the infected graft. The prognosis of arterial prosthesis infection depends greatly on early diagnosis and treatment before the onset of life-threatening complications. Knowledge of this rare association may lead to an early diagnosis and appropriate management of this serious and often fatal complication. PMID- 16721261 TI - What's new in Shock, June 2006? PMID- 16721256 TI - Group A streptococcal infections in injection drug users in Barcelona, Spain: epidemiologic, clinical, and microbiologic analysis of 3 clusters of cases from 2000 to 2003. AB - An unexplained resurgence of Group A streptococci (GAS) infections has been observed since the mid-1980s in the United States and Europe, particularly among intravenous drug users (IDUs). Several risk factors have been identified. Mutations in the capsule synthesis regulator genes (csrRS) have been associated with an increase in virulence. From January 1998 to December 2003, we conducted a prospective and retrospective descriptive analysis of invasive GAS soft-tissue infections in IDUs in Barcelona, Spain. Clinical features were collected, and we conducted a surveillance study to identify risk factors associated with GAS soft tissue infections. We analyzed chromosomal DNA by low cleavage restriction enzymes and used pulsed-field gel electrophoresis (PFGE) and variable gene sequence typing (VGST) of the emm gene to disclose the epidemiologic relationship between the strains. We analyzed the influence of clonality (M-type) and mutations in csrRS genes of these strains on clinical features. We identified 44 cases, all of which were grouped in 3 clusters: fall 2000, fall 2002, and fall 2003. Cellulitis with or without abscesses (75%) and fever (90.9%) were the most common clinical manifestations. Distant septic complications were infrequent (18.2%). Although all patients had severe infections (mainly bacteremic needle abscesses), their outcome with antibiotic therapy, usually beta-lactam, was successful in all cases. However, surgery was needed in 40.9% of patients. Through the surveillance study we found that infected patients had a higher number of drug injections per day (odds ratio [OR], 18.84; 95% confidence interval [CI], 4.83-79.4; p<0.00001), shared paraphernalia for drug use more frequently (OR, 11.11; 95% CI, 3.24-39.04; p<0.0001), were in a higher proportion both currently unemployed and homeless (OR, 4.22; 95% CI, 1.5-12.15; p<0.0001), were not in a methadone maintenance program (OR, 0.03; 95% CI, 0-0.19; p<0.00001), and more often bought drugs at a specific site (OR, 33.92; 95% CI, 7.44-174.93; p<0.00001) and from a specific dealer (OR, 72; 95% CI, 8-3090; p<0.00001), compared with patients not infected. The fall 2000 cluster was polyclonal, whereas the other 2 clusters were mainly due to the same strain of GAS (emm 25.2), and were defined as epidemic outbreaks. Clinically, the cases due to the clonal strain presented abscesses and needed surgery more frequently (p<0.001 and p=0.005, respectively). On the other hand, mutations in the csrRS genes were not associated with invasive GAS soft-tissue infection. There has been an increase in the number of cases of invasive GAS soft-tissue infections in IDUs in Barcelona, which seems to be related to drug users' habits and their socioeconomic status. Clonality (emm 25.2) but not mutations in the csrRS genes was associated with more severe GAS soft-tissue infections. PMID- 16721257 TI - Predictors of survival in systemic lupus erythematosus. AB - We conducted the current study to determine the impact of demographic factors, household income, clinical manifestations, disease activity, serologic tests, and calendar year on survival among patients with systemic lupus erythematosus (SLE). In a large prospective cohort of patients with SLE, we used the Kaplan-Meier method to estimate survival probabilities of SLE patients over time since diagnosis. We analyzed the predictors of survival in SLE using Cox proportional hazards models.The study included 1378 patients with SLE, with a median follow-up in the cohort of 6.1 years. One hundred eighteen patients died (8.6%). The overall cumulative probability of survival after disease diagnosis at 5, 10, 15, and 20 years was 95%, 91%, 85%, and 78%, respectively. Based on a multivariable model, age at SLE diagnosis >or=50 years old (hazard ratio [HR]=5.9; p<.001) and male gender (HR=2.4; p=.004) were associated with poorer survival. Patients with annual family income<25,000 dollars had poorer survival (HR=1.7; p=.040). The presence of hemolytic anemia in the first year after disease diagnosis (p=.016) or during the follow-up period (p=.031) increased the risk of death. A low complement level during the first year after diagnosis was the only serologic marker of poorer survival (p=.013 for low C3 level and p=.053 for low C4 level). PMID- 16721259 TI - Characteristics and long-term outcome of 15 episodes of systemic lupus erythematosus-associated hemophagocytic syndrome. AB - Reactive hemophagocytic syndrome (HS) occurs mainly in the setting of serious infections and lymphomas. HS can occur in the course of 2 active systemic diseases, without simultaneous infection: adult Still disease and systemic lupus erythematosus (SLE). Observations of specific lupus-associated HS are rare, and the long-term outcome of these patients with active SLE is unknown. We retrospectively studied 15 episodes of SLE-associated HS in 12 patients (10 women, 2 men) and noted the long-term outcome. HS occurred at a mean age of 25 years. All patients were febrile with >or=2 cytopenias, and bone marrow aspiration indicated hemophagocytosis. HS revealed SLE in 9 patients and recurred in 3. The main features of SLE-associated HS were a low frequency of hepatosplenomegaly, a high frequency of heart involvement (5 pericarditis, 4 myocarditis requiring transfer to intensive care unit), and a low C-reactive protein level (mean, 15 mg/L). Cutaneous-mucous symptoms of SLE, arthritis, and nephritis were present respectively in 8 (53%), 6 (40%), and 4 (27%) episodes, but symptoms of SLE were absent in 4 episodes at admission. All patients had anti nuclear antibodies when the HS occurred. Anti-double-stranded DNA antibodies were present in 12 episodes. Treatment was steroids in 14 cases but cyclophosphamide was the only treatment able to control HS in 2 cases. All the cases of SLE associated HS were controlled by the immunosuppressive regimen. Intravenous immunoglobulins seemed poorly effective. No infectious agent was found. Clinical presentations of the 23 patients with SLE-associated HS described in the literature were reviewed and were similar to those of the current series. The mean follow-up was 88 months (range, 7-240 mo). One patient died at 15 months (sepsis). Among the 5 patients with a follow-up >8 years, 4 always had active disease. During the follow-up of SLE, immunosuppressive drugs were added in 8 patients (cyclophosphamide in 7, azathioprine in 3, mycophenolate mofetil in 2) with significant adverse drug reactions. In the long-term, SLE-associated HS seems to define a severe SLE form with frequent flares, possible HS recurrences, and the need for prolonged immunosuppression. PMID- 16721262 TI - A multidisciplinary overview of cardiogenic shock. AB - Cardiogenic shock complicating acute myocardial infarction (AMI) is reviewed from multidisciplinary viewpoints encompassing both basic and clinical aspects. Insights into the absolute obligate aerobic nature of the heart which possesses neither facultative capability nor functional collateral channels, together with O2 diffusion gradients, mitochondrial O2 sensing and anaerobic ATP deficiencies, are described in some detail. Myocardial adaptive responses against energy crisis, termed the Pasteur Effect, and hypoxia inducible factor (HIF)-1 alpha are implicated for cardiomyocyte viability. Oncosis and/or lysosomal autophagy cause such overwhelming numbers (several billions) of cardiomyocyte death, virtually simultaneously following coronary thrombotic occlusion. Apoptosis is briefly described and cardiogenic shock is discussed in terms of the diagnostic criteria by MIRU, unique hemodynamic manifestations, infarct sizes and border zone extension, and potentially jeopardized myocardium in the remote areas. Reperfusion injury, i.e., reactive oxygen species (ROS), is noted as a double edged sword. The importance of early revascularization by means of PCI, CABG, and IABP support is emphasized according to current guidelines. For innovative promise in the future, de novo development of collateral channels by growth factors and trials of stem cell implantation aimed at myocardial regeneration are introduced. PMID- 16721265 TI - Aberrant acute-phase response in aged interleukin-6 knockout mice. AB - This study was designed to determine whether the acute-phase response in aged mice is altered by interleukin (IL) 6 deficiency. Young and aged wild-type (WT) and IL-6 knockout (KO) BALB/C female mice were injected with lipopolysaccharide (LPS; 1.5 microg/g body weight). After 24 h, aged IL-6 KO mice had an improved survival when compared with aged WT mice. Serum levels of IL-6 in aged WT animals given LPS were determined and, as expected, were significantly higher when compared with young LPS-treated WT animals (P<0.05). Serum levels of the acute phase protein, serum amyloid A, were 50% lower in aged LPS-treated IL-6 KO mice relative to aged WT mice given LPS (P<0.001). In contrast, the induction of LPS binding protein was not affected by age or IL-6 deficiency in LPS-treated animals. Circulating levels of corticosterone were markedly reduced in aged LPS treated IL-6 KO mice relative to aged WT mice given LPS. These data indicate that IL-6 is an important contributor to the outcome of the acute-phase response of aged individuals challenged with endotoxin. We conclude that the absence of IL-6, a cytokine that contributes to the elevated basal proinflammatory state observed in aging, can improve the ability of aged mice to withstand an otherwise lethal challenge of bacterial endotoxin. PMID- 16721264 TI - Endothelium-derived microparticles inhibit human cardiac valve endothelial cell function. AB - Elevated numbers of endothelium-derived microparticles (EMPs) in the circulation are found in a variety of critical illnesses. EMPs have been associated with vascular dysfunction, including thrombotic complications and loss of normal vascular reactivity, common responses associated with cardiac valve injury. However, the exact mechanisms of this dysfunction and the potential impact on cardiac endothelium are unknown. We hypothesize that pathologic levels of circulating EMPs negatively regulate proliferation and migration of valvular endothelial cells (ECs), leading to downstream endothelial dysfunction. EMPs were generated from plasminogen activation inhibitor 1-stimulated human umbilical vein endothelial cells (HUVECs). Human mitral valve endothelial cells (HMVECs) were isolated and characterized by platelet endothelial cell-derived adhesion molecule 1 (PECAM-1, or CD31) and von Willebrand factor immunocytochemistry. HMVECs were treated with increasing EMP doses, and then, the effects of EMPs on growth factor induced proliferation and migration were tested. Proliferation was assessed by H thymidine incorporation. EC migration was assayed by photographing microtubules of HMVECs and HUVECs in fibrin gel incubated with EMPs +/- growth factors for 48 h. The EMP effects on non-valve HUVECs were tested in parallel. EMPs inhibited HMVEC proliferation at high doses but stimulated HUVEC proliferation at all doses. In HMVECs, EMPs inhibited basic fibroblast growth factor- and vascular endothelial growth factor-induced proliferation and migration. Taken together, these data suggest EMPs regulate valvular EC proliferation in a dose-dependent manner and, furthermore, modulate growth factor signaling in ECs. These results implicate EMPs as a possible source of downstream EC dysfunction in disease states. EMPs may play a role in valvular leaflet injury in human disease by inhibiting normal growth and repair of endothelium. PMID- 16721266 TI - Dendritic cell-derived exosomes containing milk fat globule epidermal growth factor-factor VIII attenuate proinflammatory responses in sepsis. AB - In sepsis, several cell types (e.g., lymphocytes) undergo apoptosis and have the potential to harm the host if not cleared by professional phagocytes. Apoptotic cells display "eat me" signals such as phosphatidylserine that can be readily recognized by phagocytes. For full engulfment of these cells, binding to integrin alpha(v)beta(3), mediated by the bridging protein, milk fat globule epidermal growth factor-factor VIII (MFG-E8), is necessary. We hypothesized that, in sepsis, phagocytosis of apoptotic cells is impaired due to decreased MFG-E8 expression and that adoptive transfer of exosomes containing MFG-E8 is beneficial. Sepsis was induced in rats by cecal ligation and puncture (CLP) and MFG-E8 expression assessed by Western blot 20 h later. Dendritic cells were generated from bone marrow cells, and secreted exosomes were collected and injected into CLP animals. Plasma cytokines (enzyme-linked immunosorbent assay) and thymocyte apoptosis (TC-Ao, annexin V) were assessed. The ability of peritoneal macrophages from septic animals to engulf apoptotic cells was determined in an ex vivo phagocytosis assay. A 10-day survival study was conducted. Cecal ligation and puncture reduced MFG-E8 protein levels in the spleen and liver by 48% and 70%, respectively, and increased TC-Ao by 1.6-fold. Injection of MFG-E8-containing exosomes, however, led to a 33% reduced detection of TC-Ao, without directly inhibiting apoptosis. In fact, peritoneal macrophages from exosome-treated rats displayed a 2.8-fold increased ability to phagocytose apoptotic thymocytes. Inhibition of MFG-E8 before injection of exosomes completely abrogated the enhanced phagocytosis. Treatment with bone marrow dendritic cell-derived exosomes also reduced plasma tumor necrosis factor alpha and interleukin (IL)-6 levels and improved survival from 44% to 81%. We conclude that, by providing the indispensable factor MFG-E8 for complete engulfment of apoptotic cells, these exosomes lead to an attenuation of the systemic inflammatory response and overall beneficial effect in sepsis. PMID- 16721263 TI - Elevated high-mobility group box 1 levels in patients with cerebral and myocardial ischemia. AB - Cerebral and myocardial ischemia, two of the leading causes of morbidity and mortality worldwide, are associated with inflammation that can lead to multiple organ failure and death. High-mobility group box 1(HMGB1), a recently described mediator of lethal systemic inflammation, has been detected in individuals with severe sepsis and hemorrhagic shock, but its role during ischemic injury in humans is unknown. To determine whether systemic HMGB1 levels are elevated after ischemic injury, a prospective observational study was performed in subjects with a diagnosis of either Acute Coronary Syndrome (ACS) or cerebral vascular ischemia (transient ischemic attack or cerebral vascular accident). Subjects (n, 16; age [mean], 67+/-16.3 years) were enrolled in the North Shore-LIJ emergency department within 24 h of symptom onset. Blood samples were collected, and HMGB1 levels analyzed by Western blot analysis using previously described methods (Wang et al. Science. 1999). Control samples were obtained from healthy age- and sex matched volunteers (n, 16; age [mean], 68+/-15.8 years). Here, we report that serum HMGB1 levels were significantly elevated in both myocardial ischemia subjects (myocardial control serum HMGB1, 1.94+/-2.05 ng/mL, vs. myocardial ischemia serum HMGB1, 159+/-54.3 ng/mL; P<0.001); and in cerebral ischemia subjects (cerebral control serum HMGB1, 16.8+/-10.9 ng/mL, vs. cerebral ischemia serum HMGB1, 218+/-18.8 ng/mL; P<0.001). These results suggest that systemic HMGB1 levels are elevated in human ischemic disease. PMID- 16721268 TI - Glucosamine administration during resuscitation improves organ function after trauma hemorrhage. AB - Stress-induced hyperglycemia is necessary for maximal rates of survival after severe hemorrhage; however, the responsible mechanisms are not clear. One consequence of hyperglycemia is an increase in hexosamine biosynthesis, which leads to increases in levels of O-linked attachment of N-acetyl-glucosamine (O GlcNAc) on nuclear and cytoplasmic proteins. This modification has been shown to lead to improved survival of isolated cells after stress. In view of this, we hypothesized that glucosamine (GlcNH2), which more selectively increases the levels of O-GlcNAc administration after shock, will have salutary effects on organ function after trauma hemorrhage (TH). Fasted male rats that underwent midline laparotomy were bled to a mean arterial blood pressure of 40 mmHg for 90 min and then resuscitated with Ringer lactate (four times the shed blood volume). Administration of 2.5 mL of 150 mmol L GlcNH2 midway during resuscitation improved cardiac output 2-fold compared with controls that received 2.5 mL of 150 mmol L NaCl. GlcNH2 also improved perfusion of various organs systems, including kidney and brain, and attenuated the TH-induced increase in serum levels of IL-6 (902+/-224 vs. 585+/-103 pg mL) and TNF-alpha (540+/-81 vs. 345+/-110 pg mL) (values are mean+/-SD). GlcNH2 administration resulted in significant increase in protein-associated O-GlcNAc in the heart and brain after TH. Thus, GlcNH2 administered during resuscitation improves recovery from TH, as assessed by cardiac function, organ perfusion, and levels of circulating inflammatory cytokines. This protection correlates with enhanced levels of nucleocytoplasmic protein O-GlcNAcylation and suggests that increased O-GlcNAc could be the mechanism that links stress-induced hyperglycemia to improved outcomes. PMID- 16721267 TI - Proteasome-mediated regulation of CpG DNA- and peptidoglycan-induced cytokines, inflammatory genes, and mitogen-activated protein kinase activation. AB - Our previous work demonstrated that the proteasome is central to most of genes induced by lipopolysaccharide. In this study, we evaluated the role of the proteasome in response to two other microbial stimuli, CpG DNA (bacterial DNA) and peptidoglycan (PG), by measuring the effect of proteasome inhibition on cytokine secretion, induction of inflammatory gene expression, and activation of mitogen-activated protein kinases (MAPK) in murine macrophages. Pretreatment of macrophage cultures with lactacystin, a well-established proteasome inhibitor, significantly repressed tumor necrosis factor alpha secretion and tumor necrosis factor alpha and interleukin 1 beta gene expression, blocked the degradation of IkappaB, and dysregulated phosphorylation of MAPK induced by CpG DNA or PG. With respect to MAPK, lactacystin blocked expression of PG- or CpG-induced phosphorylated ERK1 and ERK2 and increased expression of phosphorylated c-Jun amino-terminal kinase but had no significant effect on phosphorylated p38. Increased expression of phoshorylated c-Jun amino-terminal kinase did not lead to an increase in AP-1 binding activity. Collectively, these data strongly support the conclusion that the proteasome is a key regulator of the CpG DNA- and PG induced signaling pathways. PMID- 16721269 TI - Neuroprotection and enhanced recovery with hypericum perforatum extract after experimental spinal cord injury in mice. AB - Oxidative stress results from an oxidant/antioxidant imbalance, an excess of oxidants, and/or a depletion of antioxidants. A considerable body of recent evidence suggests that oxidative stress and exaggerated production of reactive oxygen species play a major role in several aspects of inflammation. Hypericum perforatum is a medicinal plant species containing many polyphenolic compounds, namely, flavonoids and phenolic acids. Because polyphenolic compounds have high antioxidant potential, in this study, we evaluated the effect of H. perforatum (given at 30 mg . kg (-1)) in an experimental animal model of spinal cord injury, which was induced by the application of vascular clips to the dura via a four level T5 through T8 laminectomy. The degree of (a) spinal cord inflammation and tissue injury (histological score), (b) nitrotyrosine, (c) poly(adenosine diphosphate-ribose), (d) neutrophils infiltration, and (e) the activation of signal transducer and activator transcription 3 was markedly reduced in spinal cord tissue obtained from H. perforatum extract-treated mice. We have also demonstrated that H. perforatum extract significantly ameliorated the recovery of limb function. PMID- 16721270 TI - Decreased supply-dependent oxygen consumption in the skeletal muscle of the spontaneously hypertensive rat during acute hypoxia. AB - The purpose of this study was to correlate microvascular oxygen delivery (DO2) and consumption (VO2) in the skeletal muscle of spontaneously hypertensive rats (SHRs) and Wistar Kyoto rats (WKY) with hemodynamics during acute hypoxia. We expected greater abnormalities in central and microvascular hemodynamics during hypoxic induced shock in the SHR compared with the WKY due to microvascular rarefaction. The inspired oxygen fraction (FiO2) was lowered from 0.21 to 0.15, 0.1, 0.08, and 0.05 in anesthetized, mechanically ventilated rats. Lactate and base deficit values were similar for both groups at 0.21 and 0.15 FiO2, but higher in SHR at lower FiO2. Baseline aortic blood flow (SHR, 56.2+/-4.0 mL min; WKY, 61.8+/-5.3 mL min) and systemic DO2 (SHR, 9.02+/-0.82 mL min; WKY, 9.32+/ 0.54 mL min) increased similarly when FiO2 was lowered to 0.15. Further reductions in FiO2 caused lower aortic flow and systemic DO2 in the SHR than WKY at 0.08 and 0.05 FiO2. Spinotrapezius blood flow increased from baseline (SHR, 24.8+/-1.8 nL s; WKY, 22.7+/-2.1 nL s) in both groups when FiO2 was reduced to 0.15; further reductions in FiO2 decreased blood flow in both groups, with lower values in the SHR group at 0.1 and 0.08 FiO2. The SHR group demonstrated higher venous oxygen saturation at low values of FiO2 compared with WKY. This reduced oxygen extraction in SHR resulted in a lower supply-dependent VO2 at low values of spinotrapezius DO2, perhaps attributed to arteriolar thickening and rarefaction seen in chronic hypertension. PMID- 16721271 TI - Comparison of blood pressure and thermal responses in rats exposed to millimeter wave energy or environmental heat. AB - Electromagnetic fields at millimeter wave lengths are being developed for commercial and military use at power levels that can cause temperature increases in the skin. Previous work suggests that sustained exposure to millimeter waves causes greater heating of skin, leading to faster induction of circulatory failure than exposure to environmental heat (EH). We tested this hypothesis in three separate experiments by comparing temperature changes in skin, subcutis, and colon, and the time to reach circulatory collapse (mean arterial blood pressure, 20 mmHg) in male Sprague-Dawley rats exposed to the following conditions that produced similar rates of body core heating within each experiment: (1) EH at 42 degrees C, 35 GHz at 75 mW/cm, or 94 GHz at 75 mW/cm under ketamine and xylazine anesthesia; (2) EH at 43 degrees C, 35 GHz at 90 mW/cm, or 94 GHz at 90 mW/cm under ketamine and xylazine anesthesia; and (3) EH at 42 degrees C, 35 GHz at 90 mW/cm, or 94 GHz at 75 mW/cm under isoflurane anesthesia. In all three experiments, the rate and amount of temperature increase at the subcutis and skin surface differed significantly in the rank order of 94 GHz more than 35 GHz more than EH. The time to reach circulatory collapse was significantly less only for rats exposed to 94 GHz at 90 mW/cm, the group with the greatest rate of skin and subcutis heating of all groups in this study, compared with both the 35 GHz at 90 mW/cm and the EH at 43 degrees C groups. These data indicate that body core heating is the major determinant of induction of hemodynamic collapse, and the influence of heating of the skin and subcutis becomes significant only when a certain threshold rate of heating of these tissues is exceeded. PMID- 16721272 TI - The parp-1 inhibitor ino-1001 facilitates hemodynamic stabilization without affecting DNA repair in porcine thoracic aortic cross-clamping-induced ischemia/reperfusion. AB - Inhibition of poly (ADP-ribose) polymerase 1 (PARP-1) improved hemodynamics and organ function in various shock models induced by sepsis or ischemia/reperfusion. PARP-1, however, is also referred to play a pivotal role for the maintenance of genomic integrity. Therefore, we investigated the effect of the PARP-1 blocker INO-1001 on hemodynamics, kidney function, and DNA damage and repair during porcine thoracic aortic cross-clamping. The animals underwent 45 min of aortic cross-clamping after receiving vehicle (n=9) or i.v. INO-1001 (n=9; total dose, 4 mg.kg, administered both before clamping and during reperfusion), data were recorded before clamping, before declamping, and 2 and 4 h after declamping. During reperfusion, continuous i.v. norepinephrine was incrementally adjusted to maintain blood pressure greater than or equal to 80% of the pre-clamping level. The plasma INO-1001 levels analyzed with high-pressure liquid chromatography were 1 to 1.4 micromol/L and 0.4 to 0.6 micromol/L before and after clamping, respectively. Although INO-1001-treated animals required less norepinephrine support, kidney function was comparable in the 2 groups. There was no intergroup difference either in the time course of DNA damage and repair (comet assay) as assessed both in vivo in whole blood before surgery, before clamping, before declamping, 2 h after declamping, and ex vivo in isolated lymphocytes (Ficoll gradient) sampled immediately before clamping and analyzed before, immediately, and 1 and 2 h after exposure to 4 bar 100% O2 for 2 h. There was no difference either in the expression of the cyclin-dependent kinase inhibitor gene, p27, in the kidney (immunohistochemistry). The reduced norepinephrine requirements during reperfusion suggest a positive inotropic effect of INO-1001, as demonstrated by other authors. In our model, INO-1001 proved to be safe with respect to DNA repair. PMID- 16721273 TI - Rapid restoration of microcirculatory blood flow with hyperviscous and hyperoncotic solutions lowers the transfusion trigger in resuscitation from hemorrhagic shock. AB - Resuscitation from hemorrhagic shock relies on fluid retransfusion. However, the optimal properties of the fluid have not been established. The aim of the present study was to test the influence of the concentration of hydroxyethyl starch (HES) solution on plasma viscosity and colloid osmotic pressure (COP), systemic and microcirculatory recovery, and oxygen delivery and consumption after resuscitation, which were assessed in the hamster chamber window preparation by intravital microscopy. Awake hamsters were subjected to 50% hemorrhage and were resuscitated with 25% of the estimated blood volume with 5%, 10%, or 20% HES solution. The increase in concentration led to an increase in COP (from 20 to 70 and 194 mmHg) and viscosity (from 1.7 to 3.8 and 14.4 cP). Cardiac index and microcirculatory and metabolic recovery were improved with HES 10% and 20% when compared with 5% HES. Oxygen delivery and consumption in the dorsal skinfold chamber was more than doubled with HES 10% and 20% when compared with HES 5%. This was attributed to the beneficial effect of restored or increased plasma COP and plasma viscosity as obtained with HES 10% and 20%, leading to improved microcirculatory blood flow values early in the resuscitation period. The increase in COP led to an increase in blood volume as shown by a reduction in hematocrit. Mean arterial pressure was significantly improved in animals receiving 10% and 20% solutions. In conclusion, the present results show that the increase in the concentration of HES, leading to hyperoncotic and hyperviscous solutions, is beneficial for resuscitation from hemorrhagic shock because normalization of COP and viscosity led to a rapid recovery of microcirculatory parameters. PMID- 16721274 TI - Intranasal exposure to staphylococcal enterotoxin B elicits an acute systemic inflammatory response. AB - Staphylococcus aureus produces a variety of superantigen exotoxins, including staphylococcal enterotoxin B (SEB). Little is known regarding the pathogenesis of SEB entering through the intranasal route. Intranasal exposure to SEB might occur because of nasal packing following surgical procedure, biologic warfare, or even S. aureus colonization. We evaluated the local and systemic effects of intranasally delivered SEB using a series of human leukocyte antigen (HLA) class II transgenic mice as conventional mice expressing endogenous class II molecules mount a poor immune response to SEB. Gene expression profiling using microarrays showed robust up-regulation of genes involved in several proinflammatory pathways as early as 3 h post-intranasal challenge with SEB in HLA class II transgenic mice. This was accompanied by a several hundred-fold increase in serum levels of pro-inflammatory cytokines such as IL-12, IL-6, TNF-alpha, IFN-gamma, as well as MCP-1 in HLA class II transgenic mice but not in C57BL/6 mice; CD4 or CD8 T-cells independently contributed to the systemic cytokine response. Defective IL-12 or IL-4 receptor signaling significantly decreased or increased serum IFN-gamma, respectively. Intranasal exposure to SEB resulted in neutrophil influx into bronchoalveolar lavage fluid and caused expansion of both CD4 and CD8 T-cells expressing TCR V beta 8 in the spleen. This was accompanied by mononuclear cell infiltration in the liver reminiscent of the systemic inflammatory response syndrome. Thus, we have shown, for the first time, that intranasal administration of SEB can cause systemic immune activation. PMID- 16721275 TI - Baseline severity of sepsis in subjects of the prowess and the Kybersept clinical trials on endogenous anticoagulants. PMID- 16721278 TI - How the increase of the cervical disc space height affects the facet joint: an anatomy study. AB - STUDY DESIGN: In vitro study on the effect of increasing the height of the cervical disc space on the facet joint. OBJECTIVES: To demonstrate how facet joint articulation is affected by increasing the cervical disc space height. SUMMARY OF BACKGROUND DATA: A surgeon attempts to increase the disc space and inserts a larger artificial disc than normal in order to keep the intervertebral foramen open and the prosthesis stable. However, it is hypothesized by the current authors that this procedure could have an adverse effect on the facet joints. METHODS: Computerized tomography images passing through the disc space and the center of the C4-C7 facet joints (sagittal plane) were obtained from 15 cadaveric cervical spine specimens. A 1-mm incremental increase to a total 5 mm in disc space height was performed to simulate the changes seen in disc replacement. The change in the facet joint articulation overlap and space in the sagittal plane at normal and each displacement was measured. RESULTS: Each 1-mm incremental increase in disc space at C4-C5 translated to a decrease in the facet joint articulation overlap in the sagittal plane by approximately 8%. The mean facet joint space increased approximately 0.8 mm. At the C5-C6 and the C6-C7 levels, the articulation overlap decreased by approximately 7% and the facet joint space increased approximately 0.8 mm. CONCLUSIONS: There is a significant decrease of the facet joint articulation overlap in the sagittal plane and an increase in the facet joint space following an increase in the cervical disc space. The inappropriate increase of the disc space height may result in facet joint subluxation and could lead to the accelerated failure of the artificial disc. PMID- 16721279 TI - Pullout strength of thoracic pedicle screw instrumentation: comparison of the transpedicular and extrapedicular techniques. AB - STUDY DESIGN: In vitro biomechanical comparison of two methods of pedicle screw placement in cadaveric thoracic spine vertebrae. OBJECTIVE: Compare the biomechanical integrity of extrapedicular and transpedicular screw fixation under axial and sagittal pullout loads. SUMMARY OF BACKGROUND DATA: Extrapedicular screw placement has been advocated as a safe and effective alternative to the transpedicular screw in thoracic vertebrae. Rigorous biomechanical comparison of these two techniques is presently lacking in the literature. METHODS: Thirty seven vertebral bodies were dissected from six cadaveric thoracic spines. Each body had two polyaxial 5.0-mm screws placed: one transpedicular and one extrapedicular. The 62 screws were randomly designated for one of two loading methods: axial or sagittal. Failure load (N), taken as maximum force on the load deformation curve, and stiffness (N/mm), calculated between 50 N and 400 N, were measured. RESULTS.: Transpedicular screws were statistically stronger in both testing methods (P = 0.008). Load direction, whether axial or sagittal, had no bearing on pullout strength (P = 0.6). CONCLUSIONS: These data indicate that transpedicular screws are biomechanically superior to extrapedicular screws. This difference is small, however, and we think that extrapedicular screws offer an excellent alternative when anatomy dictates their use with other screws in segmental spinal constructs. PMID- 16721280 TI - Importance of pelvic compensation in posture and motion after posterior spinal fusion using CD instrumentation for idiopathic scoliosis. AB - STUDY DESIGN: Posture and motion analysis after a Cotrel-Dubousset instrumentation. OBJECTIVES: To investigate the compensation role of pelvis. SUMMARY OF BACKGROUND DATA: Few existing studies found no compensation at the lower unfused level but did not investigate the pelvis. METHODS: Thirty patients were analyzed before surgery then at short-, medium-, and long-term postoperative follow-up. Calibrated x-rays with three-dimensional reconstruction yielded quantification of pelvic parameters. Noninvasive optoelectronic system allowed analyzing subjects first in standing position, then during flexion-extension, lateral bending, and axial rotation. Skin markers were used to quantify three dimensional orientation of the shoulders, trunk and pelvis, and their range of motion (ROM). RESULTS: Ten patients among 21 had after surgery more than 5 degrees change of pelvic incidence. In flexion, global ROM decreased from preoperative to postoperative phase (P < 0.05). Global ROM variation was not correlated to that of lower unfused segment, while it was highly correlated to pelvic ROM variation (r = 0.78 at medium follow-up). CONCLUSION: This study underlines the central role of pelvis in balance and motion of the patients before and after surgery. PMID- 16721281 TI - Study of respiratory muscle strength, vital capacity, and ventilometry in the postoperative period of spinal surgery by posterior access. AB - STUDY DESIGN: Prospective. OBJECTIVE: To analyze tidal volume (TV), minute volume (VE), respiratory frequency (f), vital capacity (VC), maximal inspiratory (PImax), and expiratory (PEmax) pressures in patients submitted to spinal surgery for tumor or herniated disc by posterior access, and to investigate a possible association of respiratory function with surgery duration, site of surgical access, and diagnosis. SUMMARY OF BACKGROUND DATA: A reduction in pulmonary volumes is usually seen in patients submitted to thoracic or upper abdominal surgery, and recent studies have demonstrated such alterations during the postoperative period in patients submitted to craniotomy. METHODS: A total of 45 patients (mean age, 42 +/- 14 years) were submitted to spinal surgery in an University Hospital, and TV, VC, MV, f, PImax, and PEmax were measured in the preoperative period, and in the first and second postoperative days. RESULTS: Patients submitted to spinal surgery showed a decrease in the first and second postoperative days in VC, respectively (17% and 10%), TV (18% and 13%), PImax (17% and 12%), PEmax (12% and 7%), and an increase in f (18% and 12%) (P < 0.05). Reduction in TV, VC, PImax, and PEmax and the increase in f were associated with surgical time > or =240 minutes, diagnosis of tumor, and cervicothoracic surgical access (P < 0.05). CONCLUSIONS: Alterations in the respiratory function in the postoperative period of elective spinal surgery by posterior access for tumor removal or herniated disc were related to surgical time > or =240 minutes, tumor, or cervicothoracic surgical access. PMID- 16721282 TI - Blood audit evidenced-based cross-match requesting for lumbar spine surgery. AB - STUDY DESIGN: Complete audit cycle. OBJECTIVE: Radical overhaul of the blood ordering practices made by the spinal surgery department, in order to maximize the efficient use of blood products. SUMMARY OF BACKGROUND DATA: Spinal surgeons request blood-based on historical practice and not evidence. Blood is a scarce resource. Provision relies on voluntary donations from the general population. However, the donor population has been reduced because of the risks of variant Creutzfeldt-Jakob disease. Hence, the responsibility rests with clinicians to optimize their management of blood. METHODS: The spinal surgery department database was accessed to identify all patients who underwent elective lumbar spine operations (excluding deformity) during 2002 to 2003. These records were then cross-referenced with the blood transfusion department database. From the results the cross-match to transfusion (C:T) ratios and transfusion indexes (TI) were calculated for the procedures. A new tariff was agreed between the surgeons, anesthetists, and transfusion department. This was then audited prospectively. RESULTS: There were 664 cross-match requests made during 2002 to 2003, and only 40 U was transfused. This gave a C:T ratio of 16.6:1. The prospective audit resulted in only 58 U being cross-matched and 5 were transfused. This reduced the C:T ratio to 11.5:1. CONCLUSION: Implementing evidence based cross-match protocols can make significant savings in time, manpower, and money. PMID- 16721283 TI - Progressive congenital torticollis in VATER association syndrome. AB - STUDY DESIGN: A family study to reach the diagnosis of a multiple malformation syndrome. OBJECTIVE: To determine the cause of torticollis, in a patient with the VATER association. SUMMARY OF BACKGROUND DATA: The VATER association is a combination of vertebral anomalies, anal stenosis, tracheo-esophageal fistula, and radial anomalies. It needs a multidisciplinary approach with a major input from orthopedic surgeons. Torticollis in this condition has not been reported before. METHODS: Detailed family history and radiologic study using plain radiographs and three-dimensional-reconstruction. RESULTS: Bony abnormalities at the base of the skull and upper cervical vertebrae were found. CONCLUSIONS: It is postulated that the bony abnormalities were the underlying cause of the neurologic problem. PMID- 16721284 TI - Intradural cement leakage: a devastatingly rare complication of vertebroplasty. AB - STUDY DESIGN: The aim of this case report is to examine the devastating complication that may follow vertebroplasty. OBJECTIVES: To report 1 case of intradural cement leakage caused by percutaneous vertebroplasty with polymethyl methacrylate. SUMMARY OF BACKGROUND DATA: Cement leakage is not a rare complication of vertebroplasty. But intradural cement leakage is rare. We herein report a rare but devastating complication of vertebroplasty. METHODS: A 90-year old woman with a T12 and L1 osteoporotic compression fracture underwent percutaneous vertebroplasty using polymethyl methacrylate at local hospital. A literature search was performed to assess complications of vertebroplasty. RESULTS: She was transferred to our hospital due to abdominal pain. Physical examination revealed distended abdomen with local tenderness and weakness of both legs (muscle power: Grade 2). Plain radiograph of abdomen showed ileus and intradural cement leakage. Conservative treatment with nasogastric decompression was done, and her abdominal pain subsided 1 week later. CONCLUSIONS: Percutaneous vertebroplasty with polymethyl methacrylate is relatively safe, but it still should be proceeded under careful safeguard. The needle tip should not cross the medial border of the pedicle on the anteroposterior view before it has crossed the posterior cortex of the vertebral body on the lateral view. Good quality of image monitoring and clear visualization of cement should be helpful to prevent complications. PMID- 16721285 TI - Intractable radicular and low back pain secondary inferior vena cava stenosis associated with Budd-Chiari syndrome: endovascular treatment with cava stenting: case report and review of the literature. AB - STUDY DESIGN: The present report describes treatment of enlarged lumbar epidural veins in a patient with Budd-Chiari syndrome presenting with intractable radicular and low back pain. OBJECTIVE: To present a rare cause of radicular pain. To offer a case successfully to be treated with stenting of inferior vena cava stenosis. SUMMARY OF BACKGROUND DATA: Epidural venous engorgements due to inferior vena cava thrombosis have been described previously in the literature. To the authors' knowledge, this is the first case of inferior vena cava stenosis to be treated with stenting. METHODS: An 27-year-old woman presented with intractable radicular and low back pain refractory to medical treatment. She was diagnosed with Budd-Chiari syndrome for 23 years. A venography revealed severe stenosis at the hepatic portion of the inferior vena cava causing symptomatic lumbar epidural venous engorgements. RESULTS: Inferior vena cava stenosis was dilatated with endovascular stenting. Her symptoms were completely resolved after this procedure. CONCLUSIONS: Inferior vena cava stenosis related to hypertrophied caudate lobe producing lumbar epidural venous engorgements should be considered as one of the causes of radicular and low back pain. We could obtain a favorable clinical outcome by handling the primary cause of the venous engorgement. PMID- 16721287 TI - Rat spinal motion segment in organ culture: a cell viability study. AB - STUDY DESIGN: This study investigated tissue integrity and viability of cells in an organ culture system of intervertebral disc (IVD) with adjoining vertebral bodies. OBJECTIVE: The goal of this study was to design a methodology to maintain an IVD motion segment in organ culture, thereby preserving viability and tissue architecture. SUMMARY OF BACKGROUND DATA: Study of IVD mechanobiology in vitro necessitates availability of vertebral bodies for controlled application of complex loads. METHODS: IVD motion segments were dissected from rat lumbar segments and maintained in organ culture and cell viability was evaluated histochemically using NitroBlue Tetrazolium. Tissue integrity and morphology were evaluated using conventional histologic techniques. RESULTS: The in vitro organ culture of motion segments maintained the viability and tissue integrity for 14 days. More than 95% viability in all three regions of interest (anulus fibrosus, nucleus pulposus, end plates) was maintained for 14 days in culture. CONCLUSION: Our initial results suggest that long-term motion segment culture is practical, and the inclusion of vertebral bodies will facilitate anchoring during biomechanical stimulation. Thus, we expect the culture system to provide us with an excellent model for studying the pathomechanics of IVD degeneration and the effects of mechanical stimulation on the biology of IVD cells. PMID- 16721289 TI - Intervariability and intravariability of bone morphogenetic proteins in commercially available demineralized bone matrix products. AB - STUDY DESIGN: Enzyme-linked immunosorbent assay was used to detect bone morphogenetic proteins (BMPs) 2, 4, and 7 in 9 commercially available ("off the shelf") demineralized bone matrix (DBM) product formulations using 3 different manufacturer's production lots of each DBM formulation. OBJECTIVES: To evaluate and compare the quantity of BMPs among several different DBM formulations (inter product variability), as well as examine the variability of these proteins in different production lots within the same DBM formulation (intra-product variability). SUMMARY OF BACKGROUND DATA: DBMs are commonly used to augment available bone graft in spinal fusion procedures. Surgeons are presented with an ever-increasing variety of commercially available human DBMs from which to choose. Yet, there is limited information on a specific DBM product's osteoinductive efficacy, potency, and constancy. METHODS: There were protein extracts from each DBM sample separately dialyzed 4 times against distilled water at 4 degrees C for 48 hours. The amount of BMP-2, BMP-4, and BMP-7 was determined using enzyme-linked immunosorbent assay. RESULTS.: The concentrations of detected BMP-2 and BMP-7 were low for all DBM formulations, only nanograms of BMP were extracted from each gram of DBM (20.2-120.6 ng BMP-2/g DBM product; 54.2-226.8 ng BMP-7/g DBM). The variability of BMP concentrations among different lots of the same DBM formulation, intra-product variability, was higher than the variability of concentrations among different DBM formulations, inter-product variability (coefficient of variation range BMP-2 [16.34% to 76.01%], P < 0.01; BMP-7 [3.71% to 82.08%], P < 0.001). BMP-4 was undetectable. CONCLUSIONS: The relative quantities of BMPs in DBMs are low, in the order of 1 x 10(-9) g of BMP/g of DBM. There is higher variability in concentration of BMPs among 3 different lots of the same DBM formulation than among different DBM formulations. This variability questions DBM products' reliability and, possibly, efficacy in providing consistent osteoinduction. PMID- 16721291 TI - The expression of hypoxia inducible factor-1alpha and apoptosis in herniated discs. AB - STUDY DESIGN: Human intervertebral disc was obtained for the study of hypoxia inducible factor (HIF)-1alpha and apoptosis using immunohistochemical staining. OBJECTIVE: To study the expression of HIF-1alpha and apoptosis in herniated lumbar discs. SUMMARY OF BACKGROUND DATA: The presence of HIF-1alpha in human chondrocytes and rat intervertebral discs has been proven; however, to our knowledge, its expression in human intervertebral disc cells has not been reported. Apoptosis of the human intervertebral disc appears as a degenerative change caused by the aging of the intervertebral disc. To our knowledge, there is no reported study showing the correlation between apoptosis and HIF-1alpha in the human intervertebral disc. METHODS: There were 15 human intervertebral discs stained for HIF-1alpha immunohistochemically, and apoptosis was detected using the terminal deoxynucleotidyl transferase mediated-dUTP nick end labeling method. On average, the patients were 32.9 years old. The intervertebral discs were divided into noncontained (9 patients) and contained (6) groups. For the control group, 5 disc samples were used. RESULTS: The expression of HIF-1alpha was visualized in every case, with an average of 62.2% +/- 9.5% in the noncontained group, 30.5% +/- 3.6% in the contained group, and 11.4% +/- 9.3% in the control group. Apoptosis occurred in 74.3% +/- 7.3% of the cells in the noncontained group, 42.8% +/- 5.5% of the cells in the contained group, and 28% +/- 8.4% of the cells in the control group. HIF-1alpha and apoptosis expressions were both observed more frequently in the noncontained disc herniation group (P < 0.001). The correlation analysis between the degree of HIF-1alpha expression and apoptosis was also statistically significant. CONCLUSIONS: HIF-1alpha and apoptosis physiologically occur in human beings. Their expression was the highest in the noncontained group. HIF-1alpha may play a crucial role for the survival of disc cells and resorption of the herniated disc in human intervertebral discs. PMID- 16721292 TI - Mechanical modulation of vertebral growth in the fusionless treatment of progressive scoliosis in an experimental model. AB - STUDY DESIGN: Wedging of apical spinal segments was measured during creation and correction of an experimental scoliosis in a goat model. OBJECTIVES: To create and correct apical vertebral wedge deformities in a progressive experimental scoliosis model by purely mechanical means. SUMMARY OF BACKGROUND DATA: The creation and correction of vertebral wedge deformities has been previously described in a rat tail model using external fixation. METHODS: Experimental scoliosis was created in 14 goats using a posterior asymmetric tether with convex rib resection and concave rib tethering. After a period of up to 13 weeks, all tethers were removed and goats were randomized into treated (n = 8) and untreated (n = 6) groups. Treated goats underwent anterior thoracic stapling with four shape memory alloy staples along the convexity of the maximal curvature. Goats were followed for an additional 7 to 13 weeks during treatment. Serial radiographs were used to document progression or correction of the maximal scoliotic deformity as well as to measure the wedging of the apical spinal segment (two adjacent vertebrae and the intervening disc). RESULTS: During the tethering period, all goats achieved a progressive, structural, lordoscoliotic curve of significant magnitude (mean: 61 degrees, range: 49 to 73 degrees). Wedging of the apical spinal segment measured 11.1 degrees at the beginning and 22.4 degrees at the end of the tethering period. The increase in apical spinal segment wedging of +11.3 degrees (10.7 degrees vertebral/0.6-degree disc) was significant (P = 0.001). During the treatment period, the scoliosis in the stapled goats measured 56.8 degrees at the beginning and 43.4 degrees at the end for an average correction of -13.4 degrees (range: 0 to-22 degrees) (P = 0.001), whereas the untreated goats measured 67.0 degrees at the beginning and 59.8 degrees at the end for an average correction of -7.2 degrees (range: +7 to -21 degrees) (P = 0.19). Additionally, wedging of the apical spinal segment in the stapled goats measured 22.5 degrees at the beginning and 20.3 degrees at the end for an average correction of -2.2 degrees (-0.6 degrees vertebral/-1.6-degree disc); wedging of the apical vertebral segment in the untreated goats measured 22.3 degrees at the beginning and 25.8 degrees at the end of the treatment period for an average progression of +3.5 degrees (3.5 degrees vertebral/0.0-degree disc). The difference in apical spinal segment correction versus progression in the stapled (-2.2 degrees) versus control (+3.5 degrees) goats was significant (P < 0.05). CONCLUSIONS: This study demonstrates the ability to create wedge deformities at the apex of an experimental scoliosis in a large animal model and to control the progression of these deformities using anterior thoracic staples. PMID- 16721293 TI - Stress relaxation at the bone-pedicle screw interface in human bone. AB - STUDY DESIGN: A biomechanical study using human lumbar vertebra. OBJECTIVES: To assess the effects of stress relaxation on the pullout behavior of the pedicle screw in human bone. SUMMARY OF BACKGROUND DATA: The standard pullout test is widely used to assess screw-holding power; however, the effects of incremental deformation on the pullout behavior of pedicle screws are not known in human bone. A recent study in bovine bone suggests that stress relaxation during incremental pullout significantly reduces failure loads seen in pullout testing. METHODS: Fourteen human lumbar vertebrae were instrumented with pedicle screws and tested with one of two pullout protocols: standard pullout model, a continuous withdrawal, and stress relaxation pullout model, an incremental withdrawal allowing stress relaxation. Peak loads and stiffness were calculated and relaxation data were assessed using the stretched exponential function. RESULTS: Pullout strength and stiffness were significantly lower in the stress relaxation model group (P < 0.05). The characteristic relaxation time decreased and relaxation ratio increased with the increasing deformation level. The pullout and stress relaxation parameters were found to correlate with the bone quality. CONCLUSIONS: The stress relaxation process significantly affects the mechanical and viscoelastic properties of the bone-screw interface in human cadaveric bone. Stress relaxation pullout model might be a better representative of in vivo conditions by introducing the time and incremental deformation into the in vitro biomechanical testing. PMID- 16721294 TI - Segmental instability in cervical spondylotic myelopathy with severe disc degeneration. AB - STUDY DESIGN: A retrospective study was conducted. OBJECTIVE: To investigate relationships between severe disc degeneration (SDD) and segmental instability in cervical spondylotic myelopathy (CSM) and to discuss surgical treatment for CSM with severe disc degeneration. SUMMARY OF BACKGROUND DATA: Information on relationships between segmental instability and spinal cord compression in CSM with severe disc degeneration is scarce. METHODS: Radiographs and magnetic resonance images of patients with CSM with (n = 42) and without (n = 75) SDD were reviewed retrospectively. Cervical instability and spinal cord compression factors were analyzed. Outcomes of anterior cervical decompression and fusion (ACDF) and expansive laminoplasty (ELAP) were evaluated in medical records of follow-up clinics. RESULTS: Segmental instability was found in 71.4% of patients with SDD and 22.7% of patients without SDD. Spinal cord compression was found at the intervertebral space of SDD and upper adjacent disc space. The recovery rate of ACDF and ELAP was 60.8% and 57.1%, respectively. CONCLUSION: The upper adjacent vertebra above SDD has inclination of segmental instability. There is static spinal cord compression in intervertebral spaces of SDD and dynamic compression in upper adjacent intervertebral spaces. Multilevel anterior cervical decompression and fusion or expansive laminoplasty should be used for surgical treatment. PMID- 16721295 TI - Open-door laminoplasty for cervical myelopathy resulting from adjacent-segment disease in patients with previous anterior cervical decompression and fusion. AB - STUDY DESIGN: This is a retrospective study of patients with cervical myelopathy resulting from adjacent-segment disease who were treated by open-door expansive laminoplasty. OBJECTIVES: The purpose of this study was to evaluate the effectiveness of laminoplasty for cervical myelopathy resulting from adjacent segment disease. SUMMARY OF BACKGROUND DATA: Adjacent-segment disease is one of the problems associated with anterior cervical decompression and fusion. However, the optimal surgical management strategy is still controversial. METHODS: Thirty one patients who underwent open-door expansive laminoplasty for cervical myelopathy resulting from adjacent-segment disease and age- and sex-matched 31 patients with myelopathy who underwent laminoplasty as the initial surgery were enrolled in the study. The pre- and postoperative Japanese Orthopedic Association scores (JOA scores) and the recovery rate were compared between the two groups. RESULTS: The average JOA scores in the patients with adjacent-segment disease and the controls were 9.2 +/- 2.6 and 9.4 +/- 2.3 before the expansive laminoplasty and 11.9 +/- 2.8 and 13.3 +/- 1.7 at the follow-up examination, respectively; the average recovery rates in the two groups were 37.1 +/- 22.4% and 50.0 +/- 21.3%, respectively (P = 0.04). The mean number of segments covered by the high intensity lesions on the T2-weighted magnetic resonance images was 1.87 and 0.9, respectively (P = 0.001). CONCLUSIONS: Moderate neurologic recovery was obtained after open-door laminoplasty in patients with cervical myelopathy resulting from adjacent-segment disc disease, although the results were not as satisfactory as those in the control group. This may be attributed to the irreversible damage of the spinal cord caused by persistent compression at the adjacent segments. PMID- 16721296 TI - Neurologic level diagnosis of cervical stenotic myelopathy. AB - STUDY DESIGN: A cross-sectional analysis. OBJECTIVE: To elucidate the accuracy of neurologic level diagnosis of cervical stenotic myelopathy. SUMMARY OF BACKGROUND DATA: Neurologic level diagnosis in cervical myelopathy has not been well established. METHODS: A total of 106 patients with cervical stenotic myelopathy, with a single-level intramedullary high-intensity area confirmed on both preoperative and postoperative T2-weighted magnetic resonance imaging (MRI), were included in this study. We performed a level diagnosis on the basis of neurologic signs (the uppermost muscle with weakness, diminished or exaggerated deep tendon reflex, the uppermost level of sensory disturbance of the upper extremities) and compared it with a level diagnosis made by T2-weighted MRI. The sensitivity, specificity, and accuracy of neurologic signs on our index corresponding to each intervertebral level were calculated. RESULTS: The averages of sensitivity, specificity, and accuracy were 42%, 80%, and 70%, respectively, in the uppermost muscle with weakness, 66%, 89%, and 83% in deep tendon reflex, and 74%, 91%, and 87% in the sensory disturbance area. The positive and negative predictive values were 40% and 91%, respectively, in the uppermost muscle with weakness, 66% and 89% in deep tendon reflex, and 74% and 91% in the sensory disturbance area. Accuracy of a diagnosis based on muscle weakness was less high, the reason being that in many patients, the uppermost muscle with weakness was extensor digiti communis or the intrinsic muscles of the hands, and this led to a lower sensitivity. CONCLUSIONS: The average accuracy of neurologic level diagnosis based on the index we proposed was > or =70%. The level diagnosis by a sensory disturbance area showed the highest accuracy (87%). PMID- 16721297 TI - Paget sarcoma of the spine: Scottish Bone Tumor Registry experience. AB - STUDY DESIGN: Retrospective case study of 13 cases of Paget sarcoma of the spine accrued from a prospectively collected Tumor Registry database. OBJECTIVES: To analyze the clinical, radiologic, and histologic features of Paget sarcoma of the spine and to determine the factors influencing the prognosis. SUMMARY OF BACKGROUND DATA: Paget disease of bone is a common disorder with the spine being involved in over 50% of patients. However, sarcomatous degeneration in the vertebral column is an extremely rare complication. There is very little in the literature with regard to clinical presentation and prognosis of patients with Paget sarcoma affecting the vertebral column. METHODS: Between January 1944 and December 2003, 89 patients were registered with a diagnosis of Paget sarcoma in the Scottish Bone Tumor Registry. Thirteen patients with Paget sarcoma of the spine were analyzed with regard to their clinical, radiologic, and histopathologic features along with the prognostic predictors. RESULTS: The mean age was 66.9 years (range: 56-79 years). There were 10 males and three females. There were seven cases involving the sacral spine (63.6%), three cases involving lumbar vertebrae, two affecting the dorsal spine, and one with diffuse dorsolumbar involvement (D11-L3). The mode of presentation was progressively increasing low back pain (in all 13), unilateral sciatica (six; left-sided, five; right-sided, one), bilateral sciatica (two), lower limb weakness (eight), and autonomic dysfunction (four). Ten of 13 cases (76.9%) were osteosarcoma. The rest were chondrosarcoma (n = 1), fibrosarcoma (n = 1), and malignant fibrous histiocytoma (n = 1). Decompression laminectomy was performed in three patients with progressive neurologic deficit. Eight patients had received radiotherapy. The mean survival was 4.22 months. CONCLUSIONS: This series confirmed that Paget sarcoma of the spine has a very poor prognosis. We found a constellation of symptomatology in patients with sarcomatous Paget spine resulting from radiculomedullary compression, primarily lumbosacral involvement and predominantly osteosarcomatous histology. There was no significant difference observed on the overall prognosis of the patients with Paget sarcoma of the spine in the last 6 decades. PMID- 16721298 TI - Clinical outcomes of 3 fusion methods through the posterior approach in the lumbar spine. AB - STUDY DESIGN: This prospective randomized study compared 3 fusion methods: posterolateral fusion (PLF), posterior lumbar interbody fusion (PLIF), and PLIF combined with PLF (PLF+PLIF). OBJECTIVES: To compare the outcomes of the 3 fusion methods and find a useful fusion method. SUMMARY OF BACKGROUND DATA: Many studies have shown clinical results, advantages, and postoperative complications of each fusion method, but few have compared the 3 fusion methods prospectively. METHODS: A total of 167 patients who underwent 1 or 2-level fusion surgery because of degenerative lumbar disease from January 1996 to September 2000 were studied. Minimum follow-up was 3 years. The patients were randomized into 1 of 3 treatment groups: group 1 (PLF; n = 62); group 2 (PLIF; n = 57); and group 3 (PLF+PLIF; n = 48). A visual analog scale, the Oswestry Disability Questionnaire, and Kirkaldy Willis criteria were used to measure low back pain, leg pain, and disability. For radiologic evaluation, disc height, lumbar lordosis, segmental angle, and bone union were examined. Postoperative complications were also analyzed. RESULTS: At the last follow-up, good or excellent results were obtained in 50 cases of PLF (80.7%), 50 cases of PLIF (87.8%), and 41 cases of PLF+PLIF (85.5%). No statistical differences were found among the 3 groups (P = 0.704). All methods indicated significant improvement in the disc height (P < 0.05), with PLF having the highest loss in disc height. Lumbar lordosis and segmental angle increased significantly, and improvement of the segmental angle in the 3 fusion methods had statistically significant differences. The nonunion rates at the last follow-up in the 3 fusion groups were not statistically significant, with 8% in group 1, 5% in group 2, and 4% in group 3 (P > 0.05). Complications included deep infection in 3 cases, transient nerve palsy in 4, permanent nerve palsy in 1, and donor site pain in 6. CONCLUSIONS: No significant differences in clinical results and union rates were found among the 3 fusion methods. PLIF had better sagittal balance than PLF. PLIF without PLF had advantages of the elimination of donor site pain, shorter operating time, and less blood loss. PMID- 16721300 TI - Three-dimensional rotational X-ray navigation for needle guidance in percutaneous vertebroplasty: an accuracy study. AB - STUDY DESIGN: The position of a needle tip displayed on a navigation system after transpedicular introduction into a vertebral body is compared with the real position of the needle tip when using a direct navigation coupling between a three-dimensional rotational X-ray (3DRX) system and a navigation system. OBJECTIVES: To assess whether the needle tip position displayed by the navigation system corresponds to the real needle position and to quantitatively determine needle navigation accuracy in a clinically relevant setting. SUMMARY OF BACKGROUND DATA: Image-guided navigation has reportedly increased the accuracy and safety of pedicle screw insertion and decreased complication rates. In former studies, the result of image-guided navigation was mainly compared qualitatively with the result of conventional fluoroscopy-guided procedures. Previously, a direct navigation coupling between a 3DRX system and a standard navigation system was introduced that bypasses the need for explicit patient-to-image registration necessary for image-guided orthopedic surgery. In a phantom experiment, the reported accuracy of navigation with the coupling to a 3DRX system was approximately 1 mm. However, in a clinical setting, additional errors can be introduced. METHODS: Twenty-three needles were placed transpedicularly into vertebral bodies of embalmed human trunks using 3DRX-guided navigation. The navigated needle tip positions were compared with the real needle tip positions manually extracted from 3DRX volumes acquired after completion of the introduction. RESULTS: The average distance between the navigated needle tip and the real position of the needle tip extracted from a postprocedure 3DRX volume was 2.5 +/- 1.5 mm. CONCLUSIONS: Accuracy of 3DRX-guided navigation is 2.5 +/- 1.5 mm in a clinically relevant setting, which is less than the accuracy determined in phantom experiments. PMID- 16721301 TI - Magnetic resonance imaging evaluation of posterior lumbar interbody fusion. AB - STUDY DESIGN: A prospective study investigated the use of magnetic resonance imaging (MRI) scanning in the assessment of posterior lumbar interbody fusion (PLIF) using carbon fiber cages. OBJECTIVE: The aim of the study was to determine whether MRI scans in the coronal plane allow proper evaluation of interbody fusion through carbon cages. SUMMARY OF BACKGROUND DATA: Currently, there is no universally accepted radiologic assessment tool for evaluating fusion status after PLIF. METHODS: Forty-nine levels of 47 patients were studied after PLIF using 98 carbon cages with MRI scans 24 months after surgery. These images were evaluated by two independent radiologists for bridging bony trabeculation through and surrounding the cages. RESULTS: On coronal planes, the presence of bridging bony trabeculation through the cages was noted in 86 of 98 cages evaluated by observer 1 and 82 of 98 by observer 2. The bone bridging surrounding the cages was inhomogeneous. In six of 49 levels, the presence of low signal intensity zones at the cage end plate interface was observed by both radiologists, regarding three of 23 L4-L5 levels and three of 21 L5-S1 levels. CONCLUSIONS: The results of our study demonstrate that the most reliable radiographic indicator of fusion is the presence of bone bridging through the cages shown on coronal planes. PMID- 16721302 TI - Validation of a minimum outcome core set in the evaluation of patients with back pain. AB - STUDY DESIGN: Prospective study of patients with subacute osteoporotic fracture (SOF) or chronic low back pain (CLBP). OBJECTIVE: To evaluate reliability, validity, and responsiveness of a purposefully brief outcome instrument. SUMMARY OF BACKGROUND DATA: A minimum standardized "core set" was proposed for monitoring patients with low back pain in 1998, but an assessment of metric properties was still lacking. METHODS: The Core Set, SF-36, and Oswestry questionnaires were completed by 154 patients. Test-retest reproducibility was evaluated in a subsample of 43 stable patients with CLBP. Responsiveness was evaluated by estimating effect size (ES) of pre-postsurgery changes in 50 patients with SOF and 23 with CLBP. RESULTS: The total Core Set showed good reproducibility with intraclass correlation coefficients on test-retest near the highest standard of 0.9, whereas internal consistency differed between patients with CLBP and those with SOF (Cronbach's alpha of 0.92 and 0.64, respectively). Most correlations of the Core Set with SF-36 and Oswestry, previously hypothesized as high, were >0.65, demonstrating good construct validity. Sensitivity to change of the Core Set (ES 0.4-2.3) is similar to the Oswestry (ES 0.7 and 2.3). CONCLUSIONS: These findings support the potential usefulness of the Core Set when respondent burden is a major concern. However, subscale scores need to be further tested in other populations before they can be widely recommended. PMID- 16721304 TI - Cost-utility analysis of physiotherapy treatment compared with physiotherapy advice in low back pain. AB - STUDY DESIGN: Economic evaluation alongside a pragmatic multicenter randomized controlled trial from the National Health Service (NHS) and societal perspective. OBJECTIVE: To perform a cost-utility analysis of routine physiotherapy treatment compared with an assessment session and advice from a physiotherapist for patients with subacute and chronic low back pain. SUMMARY OF BACKGROUND DATA: Lack of evidence for some types of physiotherapy intervention and a paucity of cost-effectiveness data for treatment of low back pain has led to controversy and uncertainty within the medical and allied professions. PATIENTS AND METHODS: A total of 286 patients with low back pain of more than a 6-week duration were randomized to physiotherapy treatment or advice on remaining active from a physiotherapist. Data were collected on back pain-related NHS and patients' costs over a 12-month post randomization period. The primary outcome measure was the Oswestry Disability Index at 12 months, with additional Oswestry Disability Index measures at 2 and 6 months. The EuroQol EQ-5D was used to calculate quality adjusted life years. Cost-effectiveness was expressed as the incremental cost per quality adjusted life year gained. Uncertainty was handled using confidence ellipses for the ratio and cost-effectiveness acceptability curves. RESULTS: The total NHS costs were not significantly different at 179 pounds sterling (221 pounds sterling) for physiotherapy and 159 pounds sterling (260 pounds sterling) for the advice group. However, patients in the physiotherapy group had significantly higher out-of-pocket health care expenditure (40 pounds sterling, 95% confidence interval 9 pounds sterling to 71 pounds sterling). Utility levels improved in both groups from baseline to 12 months, with no significant differences between groups. CONCLUSIONS: The results indicate no significant differences in either NHS costs or effects. However, the significantly higher out of-pocket expenses incurred by patients receiving routine physiotherapy suggests that advice given by a physiotherapist should be considered as the first-line treatment for patients with this level of back pain disability. PMID- 16721305 TI - Effect of prone positioning systems on hemodynamic and cardiac function during lumbar spine surgery: an echocardiographic study. AB - STUDY DESIGN: Prospective randomized study of patients undergoing spine surgery. OBJECTIVE: To compare changes in hemodynamic and cardiac function after prone positioning using different prone positioners. SUMMARY OF BACKGROUND DATA: Prone positioning decreases blood pressure and cardiac function. Several studies have evaluated changes in cardiac function after prone positioning, and linked them to reduced venous return and ventricular compliance. This study compares different prone positioners using transesophageal echocardiography, and determines their effect on cardiac function and hemodynamics. METHODS: After correction of fluid deficits with the patient under stable anesthesia, hemodynamic and cardiac performance was measured using transesophageal echocardiography. After prone positioning, repeat measurements were performed, and comparisons were made between prone and supine positions. RESULTS: No intergroup differences in demographics, fluid deficit, baseline hemodynamics, or differences from supine to prone position were noted. Cardiac output decreased with the Wilson (Union City, CA) and Siemens AG (Munich, Germany) frames, while cardiac index and stroke volume decreased with the Andrews (Hollywood, CA), Wilson, and Siemens systems. Cardiac preload decreased using the Andrews frame. The Jackson spine table (Hollywood, CA) and bolsters had the least effect on cardiac performance. CONCLUSION: Adequate fluid replacement reduced hypotension and hemodynamic instability after prone positioning. The Jackson spine table and longitudinal bolsters had minimal effects on cardiac function, and should be considered in patients with limited cardiac reserve. PMID- 16721307 TI - A novel "four-rod technique" for lumbo-pelvic reconstruction: theory and technical considerations. AB - STUDY DESIGN: Surgical technique with case example. OBJECTIVE: To report on a novel technique that allows for the placement of 4 separate longitudinal rods across the lumbo-pelvic junction. SUMMARY OF BACKGROUND DATA: Despite advances in surgical techniques and instrumentation, lumbo-pelvic fixation remains a significant challenge. Fusions to the pelvis create long lever arms and generate high forces across the lumbosacral junction, resulting in high rates of screw pullout and implant fracture. In the attempt to achieve better bony fixation, techniques described include the use of bone cement, hydroxyapatite, and expandable screws. Although this process has decreased the incidence of screw pullout, it has not addressed the problem of rod fracture at the lumbo-pelvic junction. METHOD: There are 4 separate longitudinal rods placed across the lumbo pelvic junction that couples proximal lumbar screw anchors to 4 separate pelvic fixation points. Proximal lumbar fixation anchors are based on alternating Roy Camille "straight ahead" screws and Magerl "lateral to medial converging" pedicle screws. There are 4 distal pelvic fixation anchors used based on 1 pair of Galveston-like screws and 1 pair of proximal iliac wing screws. RESULTS: Early results of both ex vivo and in vivo reconstruction show that careful insertion of the lumbar and pelvic screws allows for divergent placement of the pedicle screw heads in a manner that 2 longitudinal rods can be placed per side, resulting in a total of 4 longitudinal rods across the lumbo-pelvic junction. Selection of cross links in various combinations allows for additional axial and torsional stability, depending on the desired reconstruction. CONCLUSION: Longer follow-up is necessary, and biomechanical and finite element studies are needed to show long-term efficacy of this technique, however, early results indicate that such a construct is feasible. Furthermore, depending on the general medical condition of the patient, immediate postoperative weight bearing is possible and reasonable. PMID- 16721309 TI - Intradiscal electrothermal therapy (IDET). PMID- 16721311 TI - Electrocardiographic changes with ziprasidone. PMID- 16721313 TI - Neuroleptic malignant syndrome and aripiprazole. PMID- 16721314 TI - The Texas Children's Medication Algorithm Project: revision of the algorithm for pharmacotherapy of attention-deficit/hyperactivity disorder. AB - OBJECTIVE: In 1998, the Texas Department of Mental Health and Mental Retardation developed algorithms for medication treatment of attention-deficit/hyperactivity disorder (ADHD). Advances in the psychopharmacology of ADHD and results of a feasibility study of algorithm use in community mental health centers caused the algorithm to be modified and updated. METHOD: We convened a consensus conference of academic clinicians and researchers, practicing clinicians, administrators, consumers, and families to revise the algorithms for the pharmacotherapy of ADHD itself as well as ADHD with specific comorbid disorders. New research was reviewed by national experts, and rationales were provided for proposed changes and additions to the algorithms. The changes to the algorithms were discussed and approved both by the national experts and experienced clinicians from the Texas public mental health system. RESULTS: The panel developed consensually agreed upon algorithms for ADHD with and without comorbid disorders. The major changes included elimination of pemoline as a treatment option, adding atomoxetine to the algorithm, and refining guidelines for treating ADHD with comorbid depression, aggressive behaviors, and tic disorders. CONCLUSIONS: Medication algorithms for ADHD can be modified to keep abreast of developments in the field. Although these evidence- and consensus-based treatment recommendations may be a useful approach to guide the treatment of ADHD in children, additional research is needed to determine how these algorithms can be used to maximally benefit child outcomes. PMID- 16721315 TI - Research-practice gaps in child mental health. AB - OBJECTIVES: To present a framework for classifying research-practice gaps to increase clarity in the discourse on evidence-informed practice and policy as it applies to child mental health. METHOD: The development of the framework was informed by the research literature about the effectiveness of clinic- and community-based interventions for the prevention and treatment of child mental health problems, patterns of uptake of these interventions, and the research literature about evidence-informed practice and policy. RESULTS: Four types of research-practice gaps are proposed: (1) the failure to implement interventions found to be effective, and the implementation of interventions (2) that have been demonstrated to cause harm, (3) that have evidence of no effect, and (4) where the effectiveness is unknown because of the lack of rigorous evaluation. Examples from child mental health are provided for each type. CONCLUSIONS: This framework could guide an agenda aimed at reducing research-practice gaps in child mental health. A range of strategies may be required to address the different types of gaps. PMID- 16721316 TI - Comparison of long-term outcomes in adolescents with anorexia nervosa treated with family therapy. AB - OBJECTIVE: To describe the relative effectiveness of a short versus long course of family-based therapy (FBT) for adolescent anorexia nervosa at long-term follow up. METHOD: This study used clinical and structured interviews to assess psychological and psychosocial outcomes of adolescents (ages 12-18 years at baseline) who were previously treated in a randomized clinical trial using family therapy between 1999 and 2002. RESULTS: Eighty-three percent (71/86) of subjects participated in follow-up assessments of current psychological and psychosocial functioning. In addition, 49% (35) were interviewed using the Eating Disorder Examination. Mean length of follow-up was 3.96 years (range 2.3-6.0 years). There were no statistically significant differences between the two groups on any measure at long-term follow-up. As a whole, the group was doing well with 89% above 90% ideal body weight, 74% with Eating Disorder Examination scores within the normal range, and 91% of postmenarcheal females not on birth control had menstrual return. CONCLUSIONS: A short course of family therapy is as effective as a longer course at follow-up. PMID- 16721317 TI - Mechanisms linking violence exposure to health risk behavior in adolescence: motivation to cope and sensation seeking. AB - OBJECTIVE: This study examined two potential mechanisms linking violence exposure and health risk behavior among adolescents in psychiatric care: sensation seeking and coping with stress through escape behavior. METHOD: Male (59%) and female adolescents (N = 251), ages 12 to 19 years, from diverse ethnic backgrounds (61% African American, 19% white, 12% Latino, 8% biracial or other ethnicity) completed a computer-administered survey assessing study variables from 1999 2004. Overall clinic consent rate was 41%. Age and gender were included in all analyses. RESULTS: Consistent with the literature on nonpsychiatric samples, violence exposure was associated with both increased substance use and sexual risk taking. Violence exposure was not associated with motivation to engage in risk behavior as a means of escape, although motivation to cope through escape was associated with a greater likelihood of substance use. Sensation seeking was related to substance use and sexual risk taking among all adolescents and with violence exposure primarily within male adolescents. CONCLUSIONS: Clinical interventions should promote adaptive coping strategies that emphasize maintaining healthy behavior, effective problem solving skills, and stress management techniques. PMID- 16721318 TI - Childhood bereavement: psychopathology in the 2 years postparental death. AB - OBJECTIVE: Although the death of a parent is one of the most significant stressors a child can experience, the psychiatric sequelae of parental death are not fully understood. METHOD: A total of 360 parent-bereaved children (ages 6-17) and their surviving parents were directly interviewed four times during the first 2 years following the death (at 2, 6, 13, and 25 months). Data collection occurred from 1989 to 1996. Psychiatric symptomatology was compared among the bereaved children, 110 depressed children, and 128 community control children and their informant parents. Additional analyses examined simple bereavement without other stressors versus complex bereavement with other stressors and anticipated versus unanticipated death. RESULTS: Bereavement following parental death is associated with increased psychiatric problems in the first 2 years after death. Bereaved children are, however, less impaired than children diagnosed with clinical depression. Higher family socioeconomic status and lower surviving parents' level of depressive symptoms are associated with better outcomes. Complex bereavement was associated with a worse course, but anticipation of the death was not. CONCLUSIONS: Childhood bereavement from parental death is a significant stressor. Children who experience depression in combination with parental depression or in the context of other family stressors are at the most risk of depression and overall psychopathology. PMID- 16721319 TI - Genetic heterogeneity between the three components of the autism spectrum: a twin study. AB - OBJECTIVE: This study investigated the etiology of autistic-like traits in the general population and the etiological overlap between the three aspects of the triad of impairments (social impairments, communication impairments, restricted repetitive behaviors and interests) that together define autism spectrum disorders. METHOD: Parents of 3,400 8-year-old twin pairs from the Twins Early Development Study completed the Childhood Asperger Syndrome Test, a screening instrument for autism spectrum symptoms in mainstream samples. Genetic model fitting of categorical and continuous data is reported. RESULTS: High heritability was found for extreme autistic-like traits (0.64-0.92 for various cutoffs) and autistic-like traits as measured on a continuum (0.78-0.81), with no significant shared environmental influences. All three subscales were highly heritable but showed low covariation. In the genetic modeling, distinct genetic influences were identified for the three components. CONCLUSIONS: These results suggest the triad of impairments that define autism spectrum disorders is heterogeneous genetically. Molecular genetic research examining the three components separately may identify different causal pathways for the three components. The analyses give no indication that different genetic processes affect extreme autistic impairments and autistic impairments as measured on a continuum, but this can only be directly tested once genes are identified. PMID- 16721320 TI - Outcome measures of Triple Board Graduates, 1991-2003. AB - OBJECTIVE: To describe program outcomes for the Combined Training Program in Child and Adolescent Psychiatry, Pediatrics, and Psychiatry (Triple Board Program). METHOD: All Triple Board Program graduates to date (1991-2003) were asked to participate in a 37-item written survey from February to April 2004. RESULTS: The response rate was 80.7%. Most graduates go on to careers in child and adolescent psychiatry, spending a mean of 72.3% of their time in the field; however, 24.8% of respondents spend at least one fourth of their time in pediatrics and 8.0% spend one fourth of their time or more in general psychiatry. Many graduates are involved in academics, teaching, and research. Board pass rates (including repeat attempts) were highest for general (95.8%) and child and adolescent psychiatry (91.4%) and lowest for pediatrics (77.2%), and a minority (36.3%) of graduates are fully "triple boarded" (i.e., have completed oral and written boards in all three fields). Of graduates, 93.8% stated they would re enroll in the Triple Board Program, and satisfaction with the curriculum was high. CONCLUSION: The Triple Board Program is successful in terms of career outcomes, involvement in academics, boards pass rates, and graduate reported satisfaction. These data may help determine the future of this combined program in addressing workforce issues in child and adolescent psychiatry. PMID- 16721321 TI - Paroxetine treatment in children and adolescents with major depressive disorder: a randomized, multicenter, double-blind, placebo-controlled trial. AB - OBJECTIVE: To assess the efficacy and tolerability of paroxetine in pediatric major depressive disorder. METHOD: Subjects 7 to 17 years old with major depressive disorder received paroxetine (10-50 mg/day) or placebo for 8 weeks from 2000 to 2001. The primary efficacy measure was change from baseline in the Children's Depression Rating Scale-Revised total score at week 8 last observation carried forward). Safety was primarily assessed by spontaneous reporting of adverse events. RESULTS: A total of 206 patients (intent to treat) were randomized to paroxetine (n = 104) or placebo (n = 102). Week 8 Children's Depression Rating Scale-Revised total score adjusted mean changes from baseline for patients receiving paroxetine and placebo were -22.58 (SE 1.47) and -23.38 points (SE 1.60), respectively (0.80, 95% confidence interval -3.09 to 4.69, p = 0.684). Increased cough (5.9% versus 2.9%), dyspepsia (5.9% versus 2.9%), vomiting (5.9% versus 2.0%), and dizziness (5.0% versus 1.0%) occurred in >or=5% of the paroxetine group and at least twice that of the placebo group. Six of 104 (5.8%) paroxetine patients reported serious adverse events compared to 1 placebo patient (1.0%). The incidence of adverse events of suicidal behavior and/or ideation while taking study medication (excluding taper) was 1.92% (2/104) for paroxetine versus 0.98% (1/102) for placebo. CONCLUSIONS: Paroxetine was not shown to be more efficacious than placebo for treating pediatric major depressive disorder. PMID- 16721322 TI - Single-dose pharmacokinetics and safety of ziprasidone in children and adolescents. AB - OBJECTIVE: The purpose of this study was to provide single-dose pharmacokinetic, safety, and tolerability data for ziprasidone in youths with tic disorder, for comparison to adult studies to discern whether ziprasidone pediatric dosing could be modeled from adult data. METHOD: A single-dose, open-label study of ziprasidone was conducted in youths (ages 7-16 years) with Tourette's disorder or chronic tic disorder. Dosing of ziprasidone oral suspension (40 mg/mL) was weight adjusted: >60 kg, 20 mg (group 1, n = 8); 31 to 60 kg, 10 mg (group 2, n = 8); and 16 to 30 kg, 5 mg (group 3, n = 8). Patients were assessed for serum ziprasidone concentration, safety, tolerability, and electrocardiogram pre- and postdose. RESULTS: Twenty-four patients were evaluated for safety and tolerability, and 23 were evaluated for pharmacokinetics. Regression analysis of AUC(0-infinity) and Cmax values versus weight-normalized dose showed linear, dose related changes in ziprasidone exposure. Ziprasidone was well tolerated with frequent, although transient, somnolence. No clinically significant change from baseline was observed in Bazett's or Fridericia's corrected QT(c) interval, and change in QT(c) interval was not related to serum ziprasidone concentration. CONCLUSIONS: Oral ziprasidone exhibited linear pharmacokinetics and dose-related exposure in youths with Tourette's disorder or chronic tic disorder, which are comparable to adult data. A single dose of ziprasidone was well tolerated without clinically significant effects on electrocardiograms collected around the time of maximum serum concentration. PMID- 16721323 TI - Early parental and child predictors of recurrent abdominal pain at school age: results of a large population-based study. AB - OBJECTIVE: To assess whether parental psychological and physical factors and child factors measured in the first year of life were associated with recurrent abdominal pain (RAP) in children at age 6(3/4) years. METHOD: A longitudinal cohort study (the Avon Longitudinal Study of Parents and Children), followed 8,272 children from pregnancy to age 6(3/4) years. Parental reports of child and parent functioning were gathered. Associations between parental and child functioning assessed at 6 to 8 months postpartum, and RAP measured at age 6(3/4) years were investigated. RESULTS: The prevalence of RAP in this sample was 11.8%. Both maternal anxiety (adjusted odds ratio = 1.53; 95% confidence interval 1.24 1.89) and paternal anxiety (adjusted odds ratio = 1.38; 95% confidence interval 1.12-1.71) in the first year of a child's life were associated with later childhood RAP. Parent reports of child temperament features such as irregular feeding and sleeping were also associated with later RAP. CONCLUSIONS: This is the first evidence from a prospective study that anxiety in both mothers and fathers and child temperament features predate the occurrence of RAP in children. These findings highlight the potential importance of addressing parental anxiety in families in which children present with RAP, although some caution should be exercised in their interpretation because of possible reporting bias. PMID- 16721324 TI - Trajectories of symptoms and impairment for pediatric patients with functional abdominal pain: a 5-year longitudinal study. AB - OBJECTIVE: This prospective study characterizes trajectories of symptoms and impairment in pediatric patients with abdominal pain not associated with identifiable organic disease. METHOD: The Children's Somatization Inventory and the Functional Disability Inventory were administered four times over 5 years to 132 patients (6-18 years old) seen in a pediatric gastroenterology clinic. Individual trajectories were empirically grouped and correlates of trajectory group membership at baseline were evaluated. RESULTS: A model with three unique trajectories was found to adequately fit both symptom and impairment data. Two trajectories indicated relatively long-term improvement and one indicated continued high levels of symptoms and impairment. At baseline, the long-term risk group did not have the most severe pain but had significantly more anxiety, depression, lower perceived self-worth, and more negative life events. CONCLUSIONS: These results indicate that several distinct trajectories of relatively long-term outcomes may exist in children with functional abdominal pain. One trajectory indicated long-term risk for a high level of symptoms and impairment. Psychosocial correlates of long-term risk for physical symptoms and impairment, such as child-reported stress and internalizing symptoms, may be useful for treatment planning. PMID- 16721325 TI - Assessing the meaning of suicidal risk behavior in adolescents: three exercises for clinicians. PMID- 16721326 TI - Getting better at getting them better: health outcomes and evidence-based practice within a system of care. PMID- 16721328 TI - Thomas Parran Award Lecture: transmission and prevention of transmission of HIV 1. PMID- 16721329 TI - Number of sexual encounters involving intercourse and the transmission of sexually transmitted infections. AB - OBJECTIVE: To investigate the impact of the number of sexual encounters involving intercourse in combination with the number of sexual partners on the transmission dynamics of sexually transmitted infections (STIs). STUDY: A statistical model is used for predicting the basic reproductive rate, R(0), which takes both the number of sexual partners and the number of sexual encounters involving intercourse per partner into consideration. The model is then applied to Swedish survey data that includes data on the number of partners over the course of 1 year, as well as the number of encounters per partner during this time. RESULTS: The number of sexual encounters involving intercourse in combination with the number of sexual partners affects the number of secondary infections in a nonlinear way. The basic reproductive rate, R(0), is lower when the heterogeneity in number of encounters is modeled than when homogeneity is assumed. CONCLUSION: The results indicate that it is possible that individuals who have a large number of partners may not, as is often assumed, be the only ones to play a central role as spreaders of STIs. Individuals who have a large number of sexual encounters involving intercourse per partner and have several (but not necessarily a very large number of) partners may also play a significant role in the transmission of STIs. PMID- 16721330 TI - Sexual bridging socially and over time: a simulation model exploring the relative effects of mixing and concurrency on viral sexually transmitted infection transmission. AB - BACKGROUND: Sexual partnerships between people at higher and lower risk for sexually transmitted infections (STIs) (i.e., bridging) occur through dissortative mixing and concurrent partnerships, yet the relative effects of these network patterns on population STI spread are poorly understood. GOAL: Using a stochastic model, the authors investigated the impact of mixing and concurrency on the spread of a persistent viral STI. STUDY DESIGN: A total of 1,050 populations were simulated of 1,000 subjects over 400 weeks with varied concurrency levels and mixing patterns. STI prevalence and the average number of secondary transmissions per subject were analyzed with regression. RESULTS: Mixing had a greater impact on prevalence for all groups, whereas concurrency was significant for only the lowest activity group. Mixing patterns moderated the magnitude of concurrency's impact on secondary transmissions. CONCLUSIONS: Through connecting subgroups of differential risk, sexual mixing facilitates dissemination of STIs throughout a population. Concurrency expedites transmission by shortening the time between sexual contacts among infected and susceptible persons, particularly during the highly infectious period. PMID- 16721331 TI - The importance of context: model projections on how microbicide impact could be affected by the underlying epidemiologic and behavioral situation in 2 African settings. AB - OBJECTIVE: The objective of this study was to explore how a microbicide's HIV impact is affected by behavioral and epidemiologic factors in 2 African settings: Cotonou, Benin, and Hillbrow, South Africa. METHODS: A mathematical model, fit to epidemiologic data from each setting, was used to estimate the HIV impact of introducing a microbicide with different HIV/sexually transmitted infection (STI) efficacies. Simulations were compared to explore how impact is affected by context. RESULTS: Widespread microbicide use results in a greater relative reduction in HIV incidence in Cotonou, where HIV/STIs are less prevalent. Most infections averted are from commercial sex in Cotonou but noncommercial sex in Hillbrow. The microbicide's STI efficacy is important in determining its HIV impact in both settings, but especially in Cotonou where the microbicide's HIV impact was mainly the result of its STI efficacy. CONCLUSIONS: It is important to develop and evaluate microbicides that are efficacious against STIs. However, even with the same patterns of use, a microbicide's impact and the importance of its STI efficacy will vary considerably between settings. PMID- 16721345 TI - Influence of post-cam design on stresses on posterior-stabilized tibial posts. AB - Polyethylene wear and fracture at the tibial post of posterior- stabilized knee prostheses are common complications. Analyzing the stresses on various designs of tibial posts should predict the risk of damage. The post-cam of knee prostheses generally are designed either as flat-on-flat or curve-on- curve contact surfaces. We investigated stress on such tibial posts using a finite element method. We applied a 500-N anteroposterior shear force to the knee at neutral contact and 10 degrees tibial internal rotation when the knee was flexed at 60 degrees , 90 degrees , 120 degrees , and 150 degrees . There was more stress during tibial rotation compared with during neutral contact. The greatest increments of maximum von Mises stress and contact stress in the flat-on-flat model were from 20.4 to 46.1 MPa (126.3%) and from 22.2 to 55.7 MPa (151%), respectively, whereas in the curve-on-curve design they were from 20.5 to 22.7 MPa (10.7%) and from 33.0 to 35.4 MPa (7.2%), respectively. Increased stress concentration at the tibial post occurred at increased flexion angles along with axial tibial rotation. The curve-on-curve design reduced stress concentration when the knee sustained an anteroposterior shear force with tibial rotation. PMID- 16721346 TI - Thigh muscle weakness in ACL-deficient knees persists without structured rehabilitation. AB - We evaluated whether quadriceps and hamstrings weakness depended on chronicity in amateur athletes with anterior cruciate ligament deficiencies. We hypothesized that the weakness would not recover to the level of healthy control subjects without structured rehabilitation. Secondarily, we asked whether quadriceps and hamstrings side-to-side percent asymmetry in strength was consistent at different stages of chronicity. Thirty-six male amateur athletes forming equal groups of short, intermediate, and long chronicity (mean, 4, 12, and 56 months, respectively) were tested isokinetically against control subjects at 60 degrees per second. Weakness was substantial in both muscle groups and at all times ranging from 32% to 21% compared with the control subjects. However, side-to-side deficits revealed a linear trend of lessening with time. The quadriceps had greater side-to-side asymmetry that ranged from 23% to 10%, whereas the hamstrings asymmetry ranged from 14% to almost 0%. Acquiring symmetric strength earlier than 1 year after injury only occurred in the hamstrings. It can be inferred that participation in organized rehabilitation would minimize the detrimental effects of anterior cruciate ligament rupture on thigh muscle strength. LEVEL OF EVIDENCE: Prognostic Study, Level II. PMID- 16721347 TI - Hybrid external fixation for arthrodesis in knee sepsis. AB - Several techniques for knee fusion have been described with success rates ranging from 29% to 100%, with worse results occurring in patients with joint sepsis. We treated 21 patients with persistent infections using knee arthrodesis with a hybrid Ilizarov frame at our institution. There were 13 men and eight women ranging from 21 to 75 years (mean, 49.7 years). Sixteen patients had chronic osteomyelitis and five had previous fusion trials. Two patients required bone transport using the same arthrodesis frame. We corrected associated malalignment in three patients. Solid knee fusion was achieved in all but one patient after a mean external fixation time of 22.7 weeks (range, 11-47 weeks). Limb shortening averaged 2.8 cm (range, 1.5-5 cm). No patients required secondary bone grafting to achieve fusion. Nine patients had complications develop, three of whom required reresection and frame application to treat persistent infection or delayed union. Our results emphasize the clinical success of using the Ilizarov fixator for knee arthrodesis after persistent sepsis. PMID- 16721348 TI - Whole patellar allograft for total knee arthroplasty after previous patellectomy. AB - We treated seven consecutive patients (nine knees) with previous total knee arthroplasties and patellectomies with whole patellar allograft reconstructions of the extensor mechanism. The patients' extensor mechanism soft tissue sleeve was intact, but the patella was not present. Deficient patellae were reconstructed using patellar ligament (whole patella) quadriceps tendon allograft. Six knees had previous patellectomies and underwent primary total knee arthroplasties. Three knees had previous patellectomies and underwent revision total knee arthroplasties. Two patients who had primary total knee arthroplasties had failed results; one from infection and one from aseptic allograft resorption and fragmentation. One patient in the revision total knee arthroplasty group had failed results from infection. After an average followup of 44 months (range, 39 48 months), the six intact knees had an increase in quadriceps strength of one grade. The average Knee Society knee and function scores increased from 59 and 63 points, respectively, to 85 and 67 points, respectfully. The average patellar thickness was 24 mm 6 weeks postoperatively, and 15 mm (38% decrease) at the most recent followup. Patellar allografting improved quadriceps function by restoring patellar height. However, a high risk of allograft complications can occur with this procedure. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series). PMID- 16721349 TI - Ambulatory activities maintain cortical bone after total hip arthroplasty. AB - Because periprosthetic bone loss impacts revision total hip arthroplasties and subsequent patient recovery, it is important to understand the consequences of stress shielding. We characterized bone loss attributed to stress shielding by investigating the influence of patient activity and demographics on reductions in cortical bone cross-sectional area, bone mineral density, and bone mineral content. Cortical bone shape, bone mineral content, and bone mineral density were measured in implanted and contralateral nonimplanted cadaveric femurs. Geometric measurements of transverse cross sections were measured with imaging software. Dual energy x-ray absorptiometry measured bone mineral content and bone mineral density. Patient activity was estimated by the mechanical usage score. The mechanical usage score was comprised of the ambulatory components of the Harris hip score. Regression analysis revealed a lower mechanical usage score and patient weight correlated with greater bone loss. Age, implant size, and time in situ did not correlate to bone loss. The implanted femurs were reduced in cross sectional area and rigidity, and expanded endosteally and periosteally. Principal axis location did not change. Bone loss was more likely to occur uniformly than to change cortical bone distribution. Bone loss was minimized in patients with higher activity levels. PMID- 16721350 TI - Low complication rate of LISS for femur fractures adjacent to stable hip or knee arthroplasty. AB - Fractures of the femur after a knee or hip arthroplasty historically have been plagued with high complication rates. The Less Invasive Stabilization System (LISS) has theoretical advantages of improved biomechanics and limited insult to the bone's vascular supply. We theorized that the LISS would have a lower complication rate than historical controls for these fractures. Patients who were treated with a LISS at two Level I trauma centers from July 2001 to July 2003 were prospectively followed up. The inclusion criteria were an acute fracture of the femur treated with a LISS in a patient with a stable ipsilateral total knee prosthesis and/or hip pros- thesis. There were 24 patients in the study group. The injury mechanism was a low-energy fall for all patients. All patients were females with an average age of 79.5 years (range. 64-93 years). Ten patients had ipsilateral hip arthroplasties, nine patients had ipsilateral total knee arthroplasties, and five patients had knee and hip arthroplasties. Followup was at an average of 48 weeks (range, 17-101 weeks). Eighteen of the 19 fractures in the surviving patients with followup healed uneventfully for a complication rate of 5.2%. One fracture was complicated by hardware pullout and was revised to a longer LISS that healed uneventfully. We think our data show that our patients had a low complication rate compared with that of historical controls, and we suggest that the LISS may be an appropriate treatment alternative for femur fractures associated with stable hip or knee prostheses. LEVEL OF EVIDENCE: Therapeutic, Level IV. PMID- 16721351 TI - A new tool for measuring cup orientation in total hip arthroplasties from plain radiographs. AB - Orientation of the hip cup is important in total hip arthroplasties. Orientation includes abduction (inclination) and anteversion. Anteversion can be considered as true (anatomic) and planar (radiographic) anteversion. Some measurement methods either are too complicated or are less precise. We developed a new protractor to measure cup orientation using postoperative anteroposterior radiographs centered at the hip. The new protractor measures true and planar anteversion and abduction easily and precisely. We verified its accuracy using a software simulator and simulated 45 radio- graphs of total hip arthroplasties with 15 different anteversions ranging from 15 degrees -29 degrees and 45 actual radiographs of total hip arthroplasties. We then measured the planar ante- version with our method and the method of Lewinnek et al. Maximal errors were 3 degrees and 2.61 degrees , respectively, and mean errors were 0.96 degrees and 1.2 degrees , respectively. The standard deviations were 0.74 degrees with our method and 0.57 degrees with the method of Lewinnek et al. For the real radiographs, the mean of absolute difference between the two methods was 1.34 degrees , and the standard deviation was 1.13 degrees . We found no difference between the two methods and no difference in our findings compared with those of Pradhan. PMID- 16721352 TI - Gorham's disease: an autopsy report. AB - We present the case report of a 35-year-old man with Gorham's disease (disappearing bone disease, massive osteolysis) with initial clinical findings of small bowel lymphangiomatosis and multicentric osteolysis. The patient, who otherwise was healthy, had a chylothorax develop and he died 9 months later of thoracic and pulmonary complications. An autopsy revealed absence of the proximal thoracic duct and significant lymphangiectatic abnormalities of the pleural, peritoneal, diaphragmatic, splenic, and small bowel tissue with lymphangiomatous masses in the thoracic and mediastinal regions. The findings suggest a subtype of Gorham's disease characterized by a dysplastic lymphatic system. Osteolysis was correlated anatomically with lymphangiectatic tissue, suggesting mediation of osteoclastic resorption via local lymphatic tissue factors. PMID- 16721353 TI - Cervical myelopathy from atlantal hypoplasia and torticollis. AB - Cervical myelopathy caused by atlantal hypoplasia has been reported in only 11 patients. Atlantal hypoplasia combined with torticollis has not been reported. Rotation of the atlantoaxial joint causes spinal canal narrowing at the atlantoaxial level and stretches the vertebral artery, which may aggravate the symptoms and signs of compressive cervical myelopathy from atlantal hypoplasia. We present a patient with cervical myelopathy from atlantal hypoplasia, retrodental pseudotumor, and torticollis. PMID- 16721354 TI - Corrective osteotomies in malunions of the distal radius: do we get what we planned? AB - Fifteen patients with symptomatic malunions of the distal radius were treated with osteotomies, corticocancellous bone grafting, and plate and screw fixation. We investigated the ability of precise preoperative planning of the size and shape of the corticocancellous bone graft to restore alignment of the radius to within 5 degrees angular deformity and 2 mm ulnar variance as compared with the opposite uninjured wrist. Only six of 15 patients (40%) satisfied these criteria. Inter-rater reliability of radiographic assessment was greater than 0.85. Five patients had residual radial inclination or sagittal tilt greater than 10 degrees with respect to the uninvolved wrist. Four patients had a residual ulnar variance greater than 2 mm with respect to the uninvolved wrist. Residual shortening (three of four patients), but not residual angulation, was associated with unsatisfactory pain and stiffness an average of 19.5 months after osteotomy (range, 11-32 months). We conclude that a distal radius osteotomy using a precisely planned and measured interpositional corticocancellous graft does not restore distal radius alignment in most patients, and that failure to restore length is associated with continued pain and stiffness. LEVEL OF EVIDENCE: Therapeutic study, Level IV (case series). PMID- 16721355 TI - Professionalism and virtues. AB - Professionalism is one of the six core competencies of both the Accreditation Council for Graduate Medical Education (ACGME) and the American Board of Medical Specialties (ABMS). It is being taught and discussed at the medical school level as well as throughout residency and continues during the lifetime of an orthopaedic surgeon. This paper will review its definition as well as the virtues that are necessary to sustain medical professionalism. PMID- 16721356 TI - Inhibition of the alpha-mannosidase Man2c1 gene expression enhances adhesion of Jurkat cells. AB - Protein N-glycosylation plays very important roles in immunity and alpha mannosidase is one of the key enzymes in N-glycosylation. This paper reports that inhibition of alpha-mannosidase Man2c1 gene expression enhances adhesion of Jurkat T cells. In comparison to the controls with normal expression of the enzyme, Jurkat cells with the inhibition of Man2c1 gene expression (AS cell) formed larger aggregates in culture, indicating an enhancement of adhesion between the cells. mRNA differential display analysis discovered up-regulation of several adhesion molecule genes in the AS cell. Because of the pivotal role played by CD54-LFA-1 interaction in immune cell interaction, this study focused on the contribution of enhanced expression of CD54 and LFA-1 to the enhanced adhesion of AS Jurkat cells. These facts, including increased binding of AS cells to ICAM-1-Fc, Mg(2+) activation of the binding of AS cells to ICAM-1-Fc and enhanced aggregation of AS cells, together with the inhibiting effect of a blocking CD11a mAb on the binding to ICAM-1-Fc and aggregation of the cells demonstrate an important contribution of enhanced CD54-LFA-1 interaction to increased adhesion between AS cells. The enhanced CD54-LFA-1 interaction also resulted in increased adhesion between AS Jurkat T cells and Raji B cells. In addition, AS cells showed cytoskeletal rearrangement. The data imply a biological significance of MAN2C1 in T-cell functioning. PMID- 16721357 TI - Regulated expression of TATA-binding protein-related factor 3 (TRF3) during early embryogenesis. AB - RNA polymerase (Pol) II transcription persists in TATA-box-binding protein (TBP)( /-) mutant mouse embryos, indicating TBP-independent mechanisms for Pol II transcription in early development. TBP-related factor 3 (TRF3) has been proposed to substitute for TBP in TBP(-/-) mouse embryos. We examined the expression of TRF3 in maturing oocytes and early embryos and found that TRF3 was co-expressed with TBP in the meiotic oocytes and early embryos from the late one-cell stage onward. The amounts of TBP and TRF3 changed dynamically and correlated well with transcriptional activity. Chromatin immunoprecipitation (ChIP) assay revealed that different gene promoters in mouse embryonic stem (ES) cells recruited TRF3 and TBP selectively. Comparative analyses of TRF3 and TBP during cell cycle showed that both factors proceeded through cell cycle in a similar pace, except that TRF3 was slightly delayed than TBP in entering the nucleus when cells were exiting the M-phase. Data from expression and biochemical analyses therefore support the hypothesis that TRF3 plays a role in early mouse development. In addition, results from co-localization study suggest that TRF3 may be also involved in Pol I transcription. PMID- 16721358 TI - Phase I/II trial of doxorubicin and fixed dose-rate infusion gemcitabine in advanced soft tissue sarcomas: a GEIS study. AB - The aim of the study was to determine the dose-limiting toxicity and maximum tolerated dose of a first-line combination of doxorubicin and gemcitabine in adult patients with advanced soft tissue sarcomas and to explore its activity and toxicity, and the presence of possible interactions between these agents. Patients with measurable disease were initially treated with doxorubicin 60 mg m( 2) by i.v. bolus on day 1 followed by gemcitabine at 800 mg m(-2) over 80 min on days 1 and 8, every 21 days. Concentrations of gemcitabine and 2',2' difluorodeoxyuridine in plasma, and gemcitabine triphosphate levels in peripheral blood mononuclear cells were determined during 8 h after the start of gemcitabine infusion. Myelosuppression and stomatitis were limiting toxicities, and the initial dose level was applied for the Phase II trial, where grade 3-4 granulocytopenia occurred in 70% of patients, grade 3 stomatitis in 46% and febrile neutropenia in 20%. Objective activity in 36 patients was 22% (95% CI: 9 35%), and a 50% remission rate was noted in leiomyosarcomas. Administration of doxorubicin preceding gemcitabine significantly reduced the synthesis of gemcitabine triphosphate. Clinical activity, similar to that of single-agent doxorubicin, and the toxicity encountered do not justify further studies with this schedule of administration. PMID- 16721359 TI - Angiogenesis in a human neuroblastoma xenograft model: mechanisms and inhibition by tumour-derived interferon-gamma. AB - Tumour progression in neuroblastoma (NB) patients correlates with high vascular index. We have previously shown that the ACN NB cell line is tumorigenic and angiogenic in immunodeficient mice, and that interferon-gamma (IFN-gamma) gene transfer dampens ACN tumorigenicity. As IFN-gamma represses lymphocyte-induced tumour angiogenesis in various murine models and inhibits proliferation and migration of human endothelial cells, we have investigated the antiangiogenic activity of tumour-derived IFN-gamma and the underlying mechanism(s). In addition, we characterised the tumour vasculature of the ACN xenografts, using the chick embryo chorioallantoic membrane assay. We show that the ACN/IFN-gamma xenografts had a lower microvessel density and less in vivo angiogenic potential than the vector-transfected ACN/neo. The vascular channels of both xenografts were formed by a mixed endothelial cell population of murine and human origin, as assessed by the FICTION (fluorescence immunophenotyping and interphase cytogenetics) technique. With respect to ACN/neo, the ACN/IFN-gamma xenografts showed more terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling-positive human and murine endothelial cells, suggesting that inhibition of angiogenesis by IFN-gamma was dependent on the induction of apoptosis, likely mediated by nitric oxide. Once the dual origin of tumour vasculature is confirmed in NB patients, the xenograft model described here will prove useful in testing the efficacy of different antiangiogenic compounds. PMID- 16721360 TI - The presence of a systemic inflammatory response predicts poorer survival in patients receiving adjuvant 5-FU chemotherapy following potentially curative resection for colorectal cancer. AB - There is increasing evidence that the presence of a systemic inflammatory response plays an important role in survival following curative resection for colorectal cancer. The present study evaluated the relationship between C reactive protein concentrations and survival in a cohort of patients receiving adjuvant 5-fluorouracil (5-FU) chemotherapy following potentially curative resection for colorectal cancer. In all, 222 patients undergoing potentially curative resection for colorectal cancer were studied. Of these, 50 patients received adjuvant 5-FU-based chemotherapy. Circulating concentrations of C reactive protein were measured prior to surgery. The minimum follow-up was 15 months; the median follow-up of the survivors was 38 months. During this period 61 patients died, 32 patients of their cancer and 29 of intercurrent disease. In those patients who did not receive adjuvant chemotherapy, age (P < 0.001), Dukes stage (P < 0.05) and an elevated C-reactive protein (P < 0.01) were significantly associated with survival. In those patients who did receive adjuvant chemotherapy, an elevated C-reactive protein concentration (P < 0.01) was significantly associated with survival. The presence of a systemic inflammatory response is an independent predictor of poor outcome in patients receiving adjuvant 5-FU-based chemotherapy following potentially curative resection for colorectal cancer. PMID- 16721361 TI - Expression and localisation of Akt-1, Akt-2 and Akt-3 correlate with clinical outcome of prostate cancer patients. AB - We investigated the correlation between the expression and localisation of Akt-1, Akt-2, Akt-3, phospho-Akt proteins and the clinicopathological parameters in 63 prostate cancer specimens. More than 60% of cancerous tissues overexpressed Akt 1, Akt-2 or Akt-3. Cytoplasmic Akt-1 expression was correlated with a higher risk of postoperative prostate-specific antigen (PSA) recurrence and shorter PSA recurrence interval. Cytoplasmic Akt-2 did not show any significant correlation with clinicopathological parameters predicting outcomes. Cytoplasmic Akt-3 was associated with hormone-refractory disease progression and extracapsular invasion. Nuclear Akt-1 and Akt-2 expression were correlated with favourable outcome parameters such as absence of lymph node and perineural invasion. Kaplan Meier analysis and Cox regression model also showed that Akt-1 and Akt-2, but not Akt-3 or phospho-Akt was associated with a significantly higher risk of PSA recurrence. In contrast, nuclear Akt-1 was significantly associated with a lower risk of PSA recurrence. Multivariate analysis revealed that clinical stage, Gleason score and the combined cytoplasmic nuclear Akt-1 marker in cancerous tissues were significant independent prognostic factors of PSA recurrence. This is the first report demonstrating in patients with prostate cancer and the particular role of Akt-1 isoform expression as a prognostic marker depending of its localisation. PMID- 16721362 TI - A loss of c-kit expression is associated with an advanced stage and poor prognosis in breast cancer. AB - To evaluate the c-kit expression in breast cancer, 217 invasive ductal carcinomas of the breast were immunohistochemically stained for c-kit protein. The c-kit expression was positive in 59 (27%) of 217 tumours, while the c-kit expression was negative in 158 (73%) of 217 tumours. There was a significant correlation between a negative expression of the c-kit protein and lymph node metastasis (P < 0.0001), and the incidence of a negative expression of the c-kit protein increased as the number of the metastatic lymph nodes increased (P = 0.0003). The c-kit expression did not significantly correlate with the tumour size, nuclear grade, oestrogen receptor status, MIB-1 counts and p53 protein expression. A univariate analysis indicated the patients with the negative c-kit expression to have a worse disease-free survival (DFS) than those with the positive c-kit expression (P = 0.0041), while a multivariate analysis determined lymph node metastases and the MIB-1 counts to be independently significant factors for DFS. In conclusion, a loss of the c-kit expression was found in about three-fourth of invasive ductal carcinoma of the breast and was associated with lymph node metastases. The prognostic implications of the c-kit expression seem to be due to fact that a loss of the c-kit expression is associated with an advanced stage of breast cancer. PMID- 16721363 TI - Reduced tau expression in gastric cancer can identify candidates for successful Paclitaxel treatment. AB - A recent study disclosed that breast cancer cases with low 'tau' expression can predict susceptibility to Paclitaxel administration. In the current study, the clinical significance of tau expression in gastric cancer cases was established by identifying candidates with Paclitaxel administration. Tissue specimens from 20 cases of in-operable or noncuratively resected gastric cancer were examined. Subsequent to the administration of 80 mg m(-2) of Paclitaxel in six 3-h intravenous infusions, the clinical effectiveness of Paclitaxel was evaluated by the size of metastatic lesions with computed tomography. The status of the tau expression was determined by immunohistochemistry. Based on a previously reported classification scheme, six were classified as tau-negative expression (0, 1+) cases and 14 were classified as tau-positive expression (2+, 3+) cases. All six (100%) cases of tau-negative expression showed a favourable response (partial response or minor response) to Paclitaxel administration. However, 12 (86%) of the 14 cases of tau-positive expression showed progressive disease (n = 11) or no change (n = 1) after Paclitaxel administration. The serum carcinoembryonic antigen values of the six cases of tau-negative expression were markedly decreased in comparison to the 14 tau-positive cases. These data indicate that tau-negative expression can be used to select gastric cancer patients, which will favourably respond to Paclitaxel treatment. PMID- 16721364 TI - FGFR4 Arg388 allele correlates with tumour thickness and FGFR4 protein expression with survival of melanoma patients. AB - A single nucleotide polymorphism in the gene for FGFR4 (-Arg388) has been associated with progression in various types of human cancer. Although fibroblast growth factors (FGFs) belong to the most important growth factors in melanoma, expression of FGF receptor subtype 4 has not been investigated yet. In this study, the protein expression of this receptor was analysed in 137 melanoma tissues of different progression stages by immunohistochemistry. FGFR4 protein was expressed in 45% of the specimens and correlated with pTNM tumour stages (UICC, P = 0.023 and AJCC, P = 0.046), presence of microulceration (P = 0.009), tumour vascularity (P = 0.001), metastases (P = 0.025), number of primary tumours (P = 0.022), overall survival (P = 0.047) and disease-free survival (P = 0.024). Furthermore, FGFR4 Arg388 polymorphism was analysed in 185 melanoma patients by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The Arg388 allele was detected in 45% of the melanoma patients and was significantly associated with tumour thickness (by Clark's level of invasion (P = 0.004) and by Breslow in mm (P = 0.02)) and the tumour subtype nodular melanoma (P = 0.002). However, there was no correlation of the FGFR4 Arg388 allele with overall and disease-free survival. In conclusion, the Arg388 genotype and the protein expression of FGFR4 may be potential markers for progression of melanoma. PMID- 16721365 TI - Increased expression of pAKT is associated with radiation resistance in cervical cancer. AB - Phosphorylated AKT (pAKT) is a major contributor to radioresistance in human cancers. The aim of this study was to investigate the association of pAKT expression and radiation resistance in cervical cancer. A retrospective review was made of the records of 27 women who received primary radiation therapy due to locally advanced cervical cancer (LACC) with FIGO stage IIB-IVA. Nine patients regarded as radiation resistant developed local recurrences with a median progression free interval of 9 months. Eighteen patients did not show local recurrences, and were regarded as a radiation-sensitive group. Using pretreatment paraffin-embedded tissues, we evaluated pAKT expression by immunohistochemistry. A significant association was found between the level of pAKT expression and local recurrence. Immunohistochemical staining for pAKT was significantly more frequent in the radiation-resistant than in the radiation-sensitive group (P=0.004). The mean progression-free survival was 86 months for patients with pAKT-negative staining (19 cases) and 44 months for patients with pAKT-positive expression (eight cases) (P=0.008). These results suggest that signalling from phosphatidylinositide 3-kinase/pAKT can lead to radiation resistance, and that evaluation of pAKT may be a prognostic marker for response to radiotherapy in LACC. PMID- 16721366 TI - Real-time PCR analysis of a 3895 bp mitochondrial DNA deletion in nonmelanoma skin cancer and its use as a quantitative marker for sunlight exposure in human skin. AB - Previous findings from our own laboratory have shown that the frequency of occurrence (i.e. the simple presence or absence) of the 3895 bp mitochondrial DNA deletion is increased with increasing sun exposure. The present study has significantly extended this work by developing, validating and then using a quantitative real-time PCR assay to investigate for the first time the actual level (as opposed to the frequency of occurrence) of the 3895 bp deletion in human skin from different sun-exposed body sites and tumours from nonmelanoma skin cancer patients. We investigated the 3895 bp deletion in 104 age-matched split human skin samples taken from various sun-exposed sites defined as usually exposed (n = 60) and occasionally exposed (n = 44) when outdoors. The results clearly show an increased level of the 3895 bp deletion with increasing sun exposure. Specifically, there was a significantly higher level of the deletion in the usually sun-exposed compared to the occasionally sun-exposed skin (P = 0.0009 for dermis, P = 0.008 for epidermis; two-tailed t-test). Our study has also extended previous findings by showing that the level of the 3895 bp deletion is significantly higher in the dermis compared with the epidermis both in the occasionally sun-exposed samples (P = 0.0143) and in the usually sun-exposed skin. (P = 0.0007). PMID- 16721367 TI - Production and upregulation of granulocyte chemotactic protein-2/CXCL6 by IL 1beta and hypoxia in small cell lung cancer. AB - Small cell lung cancer (SCLC) is characterised by early and widespread metastasis. However, SCLC cells have so far been found to produce low levels of known pro-angiogenic factors. We speculated that SCLC cells might produce alternative pro-angiogenic factors. Here, we report that a panel of SCLC cell lines constitutively secrete granulocyte chemotactic protein-2 (GCP-2)/CXCL6, a CXC ELR+ chemokine. In contrast, none of the three tested NSCLC cell lines secreted GCP-2. Production of GCP-2 in vivo was also confirmed in seven out of nine specimens with SCLC. We demonstrate that expression of GCP-2 is mediated by NF-kappaB as ALLN, an NF-kappaB pathway inhibitor, almost completely abolished GCP-2 production in SCLC cell lines. We also demonstrate that GCP-2 can be significantly upregulated by IL-1beta and hypoxia in SCLC cell lines. This result suggests a role for GCP-2 in promoting tumour progression in vivo under unfavourable conditions such as oxygen deprivation. As SCLC cells express both GCP-2 and its receptors CXCR1 and CXCR2, their biological significance in SCLC progression was further studied. We demonstrate that GCP-2 is an autocrine growth factor. Cell proliferation was significantly inhibited by anti-GCP-2 neutralising antibody in two high-GCP-2-producing cell lines. In addition, expression of the proliferation marker PCNA was upregulated by exogenous GCP-2 in two low-GCP-2 producing cell lines. Taken together, these results suggest an important role for GCP-2 as an autocrine mitogen in the growth and metastasis of SCLC. PMID- 16721368 TI - Increasing incidence of squamous cell carcinoma of the anus in Scotland, 1975 2002. AB - In Scotland, since 1975-1979 (world) age-standardised incidence of squamous cell carcinoma of the anus has more than doubled, reaching 0.37 per 100,000 in males and 0.55 in females during 1998-2002, being somewhat higher in socioeconomically deprived areas. PMID- 16721369 TI - A phase I/II multicentric trial of gemcitabine and epirubicin in patients with advanced pancreatic carcinoma. AB - Potential synergistic interaction between gemcitabine (GEM) and epirubicin (EPI) in pancreatic cancer have been described previously. The maximum-tolerated dose in this trial was GEM 1000 mg m(-2) and EPI 45 mg m(-2). Median time to progression was 5.1 months and median survival time 7.4 months. This combination appears well tolerated and shows promising clinical activity. PMID- 16721370 TI - Multiple primary tumours in women following breast cancer, 1973-2000. AB - We investigated the predictors of the risk of developing a second primary cancer after breast cancer, this occurring in about 12% of affected women. The analysis included 335 191 females, registered in the National Cancer Institute's Surveillance Epidemiology and End Results (SEER) database, who had been diagnosed with breast cancer. Observed numbers of subsequent cancers in the SEER database with a first breast cancer diagnosed from 1973 to 2000 were compared with the expected numbers based on age-adjusted incidence rates to calculate standardised incidence ratios. Kaplan-Meier curves were conducted to determine the median time until the second primary cancer diagnosis. Average number of years until diagnosis varied by site and by age as well as median years until second cancer diagnosis. Most cancer risks decreased with age, but there was an increase in aging-related cancers such as lung cancer. The median years of follow-up were well beyond the 5-year mark. Breast cancer survivors should be advised of their increased risk for developing certain cancers in their lifetime. PMID- 16721371 TI - AMN107 (nilotinib): a novel and selective inhibitor of BCR-ABL. AB - Chronic myelogenous leukaemia (CML) and Philadelphia chromosome positive (Ph+) acute lymphoblastic leukaemia (ALL) are caused by the BCR-ABL oncogene. Imatinib inhibits the tyrosine kinase activity of the BCR-ABL protein and is an effective, frontline therapy for chronic-phase CML. However, accelerated or blast-crisis phase CML patients and Ph+ ALL patients often relapse due to drug resistance resulting from the emergence of imatinib-resistant point mutations within the BCR ABL tyrosine kinase domain. This has stimulated the development of new kinase inhibitors that are able to over-ride resistance to imatinib. The novel, selective BCR-ABL inhibitor, AMN107, was designed to fit into the ATP-binding site of the BCR-ABL protein with higher affinity than imatinib. In addition to being more potent than imatinib (IC50< 30 nM) against wild-type BCR-ABL, AMN107 is also significantly active against 32/33 imatinib-resistant BCR-ABL mutants. In preclinical studies, AMN107 demonstrated activity in vitro and in vivo against wild-type and imatinib-resistant BCR-ABL-expressing cells. In phase I/II clinical trials, AMN107 has produced haematological and cytogenetic responses in CML patients, who either did not initially respond to imatinib or developed imatinib resistance. Dasatinib (BMS-354825), which inhibits Abl and Src family kinases, is another promising new clinical candidate for CML that has shown good efficacy in CML patients. In this review, the early characterisation and development of AMN107 is discussed, as is the current status of AMN107 in clinical trials for imatinib-resistant CML and Ph+ ALL. Future trends investigating prediction of mechanisms of resistance to AMN107, and how and where AMN107 is expected to fit into the overall picture for treatment of early-phase CML and imatinib-refractory and late-stage disease are discussed. PMID- 16721372 TI - Phase II trial of oral S-1 combined with gemcitabine in metastatic pancreatic cancer. AB - We conducted a phase II trial of gemcitabine with S-1, oral fluorouracil (5-FU) prodrug tegafur combined with two modulators, 5-chloro-2, 4-dihydroxypyridine and potassium oxonate, to evaluate the activity and toxicity of such a combination in metastatic pancreatic cancer (MPC) patients. Patients who had pathologically proven pancreatic cancer with metastatic lesions were eligible candidates for entry into the study. S-1 was given orally (30 mg m(-2)) b.i.d. for 14 consecutive days and gemcitabine (1000 mg m(-2)) was given on days 8 and 15. The cycle was repeated every 21 days. We enrolled 33 MPC patients. The median number of cycles was eight (range 1-20). Grade 3-4 toxicities were leucopenia (33%), neutropenia (55%), anaemia (9%), thrombocytopenia (15%), anorexia (6%), fever (9%), and interstitial pneumonia (6%). Objective responses were obtained in 16 patients (one complete response and 15 partial responses; response rate, 48%; 95% confidence interval (CI), 33-65). Median survival and 1-year survival rate were 12.5 months (95% CI, 5.9-19.1) and 54% (95% CI, 36-72), respectively. Combination chemotherapy with GEM and S-1 was well tolerated and yielded a significantly high response rate. PMID- 16721373 TI - Growth inhibition and differentiation of human breast cancer cells by the PAFR antagonist WEB-2086. AB - WEB-2086 -- an antagonist of platelet-activating factor receptor (PAFR) with known anti-inflammatory, antiangiogenic and antileukaemic properties -- also proved to inhibit the proliferation in human solid tumour cell lines of different histology, and with much higher efficacy than in normal fibroblasts. A detailed analysis of WEB-2086 anticancer activity was then performed focusing on breast adenocarcinoma MCF-7 and MDA-MB-231 cells. WEB-2086-treated cells, either expressing (MCF-7) or unexpressing (MDA-MB-231) the oestrogen receptor (ER)alpha, underwent a dose-dependent growth arrest (IC(50)=0.65+/-0.09 and 0.41+/-0.07 mM, respectively) and accumulation in G(0)-G(1) phase. WEB-2086 also induced morphological and functional changes typical of mature mammary phenotype including (i) cell enlargement and massive neutral lipid deposition (best accomplished in MCF-7 cells); (ii) decrease in motility and active cathepsin D levels (mainly observed in highly invasive MDA-MB-231 cells). The expression of ERalpha was neither increased nor reactivated in treated MCF-7 or MDA-MB-231 cells, respectively. WEB-2086-induced differentiation in breast cancer cells involved the upregulation of PTEN, a key tumour suppressor protein opposing tumorigenesis, and was apparently independent of p53, PAFR, peripheral benzodiazepine receptor and ERalpha status. Overall, WEB-2086 can be proposed as an effective antiproliferative and differentiative agent with interesting translational opportunities to treat breast cancers in support to conventional chemotherapy. PMID- 16721374 TI - A solution to the problem of ion confounding in experimental biology. PMID- 16721375 TI - Revealing the hidden structure of our genome. PMID- 16721376 TI - Protein ligation: an enabling technology for the biophysical analysis of proteins. AB - Biophysical techniques such as fluorescence spectroscopy and nuclear magnetic resonance (NMR) spectroscopy provide a window into the inner workings of proteins. These approaches make use of probes that can either be naturally present within the protein or introduced through a labeling procedure. In general, the more control one has over the type, location and number of probes in a protein, then the more information one can extract from a given biophysical analysis. Recently, two related approaches have emerged that allow proteins to be labeled with a broad range of physical probes. Expressed protein ligation (EPL) and protein trans-splicing (PTS) are both intein-based approaches that permit the assembly of a protein from smaller synthetic and/or recombinant pieces. Here we provide some guidelines for the use of EPL and PTS, and highlight how the dovetailing of these new protein chemistry methods with standard biophysical techniques has improved our ability to interrogate protein function, structure and folding. PMID- 16721378 TI - Interrogation of genomes by molecular copy-number counting (MCC). AB - Human cancers and some congenital traits are characterized by cytogenetic aberrations including translocations, amplifications, duplications or deletions that can involve gain or loss of genetic material. We have developed a simple method to precisely delineate such regions with known or cryptic genomic alterations. Molecular copy-number counting (MCC) uses PCR to interrogate miniscule amounts of genomic DNA and allows progressive delineation of DNA content to within a few hundred base pairs of a genomic alteration. As an example, we have located the junctions of a recurrent nonreciprocal translocation between chromosomes 3 and 5 in human renal cell carcinoma, facilitating cloning of the breakpoint without recourse to genomic libraries. The analysis also revealed additional cryptic chromosomal changes close to the translocation junction. MCC is a fast and flexible method for characterizing a wide range of chromosomal aberrations. PMID- 16721377 TI - Assaying chromosomal inversions by single-molecule haplotyping. AB - Inversions are an important form of structural variation, but they are difficult to characterize, as their breakpoints often fall within inverted repeats. We have developed a method called 'haplotype fusion' in which an inversion breakpoint is genotyped by performing fusion PCR on single molecules of human genomic DNA. Fusing single-copy sequences bracketing an inversion breakpoint generates orientation-specific PCR products, exemplified by a genotyping assay for the int22 hemophilia A inversion on Xq28. Furthermore, we demonstrated that inversion events with breakpoints embedded within long (>100 kb) inverted repeats can be genotyped by haplotype-fusion PCR followed by bead-based single-molecule haplotyping on repeat-specific markers bracketing the inversion breakpoint. We illustrate this method by genotyping a Yp paracentric inversion sponsored by >300 kb-long inverted repeats. The generality of our methods to survey for, and genotype chromosomal inversions should help our understanding of the contribution of inversions to genomic variation, inherited diseases and cancer. PMID- 16721379 TI - Region-directed phototransfection reveals the functional significance of a dendritically synthesized transcription factor. AB - Multiple nuclear transcription factors including E-26-like protein 1 (Elk-1) have been found in neuronal dendrites, yet the functional significance of such localization has not yet been explained. Here we use a focal transfection procedure, 'phototransfection', to introduce Elk1 mRNA into specific regions of live, intact primary rat neurons. Introduction and translation of Elk1 mRNA in dendrites produced cell death, whereas introduction and translation of Elk1 mRNA in cell bodies did not produce cell death. Elk-1 translated in dendrites was transported to the nucleus, and cell death depended upon transcription, supporting the dendritic imprinting hypothesis and highlighting the importance of the dendritic environment on protein function. Our demonstration of the utility of phototransfection for spatially controlled introduction of mRNAs opens the broader opportunity to use this method to introduce selected quantities of small molecules into discrete regions of live cells to assess their biological functions. PMID- 16721380 TI - Site-specific recombination in human embryonic stem cells induced by cell permeant Cre recombinase. AB - The biomedical application of human embryonic stem (hES) cells will increasingly depend on the availability of technologies for highly controlled genetic modification. In mouse genetics, conditional mutagenesis using site-specific recombinases has become an invaluable tool for gain- and loss-of-function studies. Here we report highly efficient Cre-mediated recombination of a chromosomally integrated loxP-modified allele in hES cells and hES cell-derived neural precursors by protein transduction. Recombinant modified Cre recombinase protein translocates into the cytoplasm and nucleus of hES cells and subsequently induces recombination in virtually 100% of the cells. Cre-transduced hES cells maintain the expression of pluripotency markers as well as the capability of differentiating into derivatives of all three germ layers in vitro and in vivo. We expect this technology to provide an important technical basis for analyzing complex genetic networks underlying human development as well as generating highly purified, transplantable hES cell-derived cells for regenerative medicine. PMID- 16721381 TI - SuperSAGE array: the direct use of 26-base-pair transcript tags in oligonucleotide arrays. AB - We developed a new platform for genome-wide gene expression analysis in any eukaryotic organism, which we called SuperSAGE array. The SuperSAGE array is a microarray onto which 26-bp oligonucleotides corresponding to SuperSAGE tag sequences are directly synthesized. A SuperSAGE array combines the advantages of the highly quantitative SuperSAGE expression analysis with the high-throughput microarray technology. We demonstrated highly reproducible gene expression profiling by the SuperSAGE array for 1,000 genes (tags) in rice. We also applied this technology to the detailed study of expressed genes identified by SuperSAGE in Nicotiana benthamiana, an organism for which sufficient genome sequence information is not available. We propose that the SuperSAGE array system represents a new paradigm for microarray construction, as no genomic or cDNA sequence data are required for its preparation. PMID- 16721383 TI - Inflammation and multiple myeloma: the Toll connection. PMID- 16721384 TI - Clone wars in CML. PMID- 16721385 TI - Molecular profiling reveals myeloid leukemia cell lines to be faithful model systems characterized by distinct genomic aberrations. AB - To model and investigate different facets of leukemia pathogenesis, a widely accepted approach is to use immortalized leukemia cell lines. Although these provide powerful tools to our knowledge, few studies have addressed the question whether hematopoietic cell lines represent accurate and reliable model systems. To improve the molecular characterization of these model systems, we analyzed 17 myeloid leukemia cell lines using DNA microarray technology. By array-based comparative genomic hybridization, we identified recurrent genomic DNA gains and losses, as well as high-level amplifications. Parallel analysis of gene expression helped delineate potential candidate genes, and unsupervised analysis of gene expression data revealed cell lines to cluster in part based on underlying cytogenetic abnormalities. Comparison with clinical leukemia specimens showed that key signatures were retained, as myeloid cell lines with characteristic cytogenetic aberrations co-clustered with leukemia samples carrying the respective abnormality. Signatures were also quite robust, as expression data from cell lines correlated highly with published data. Thus, our analyses demonstrate myeloid cell lines to exhibit conserved and stable signatures reflecting the underlying primary cytogenetic aberrations. Our refined molecular characterization of myeloid cell lines supports the utility of cell lines as faithful and powerful model systems and provides additional insights into the molecular mechanisms of leukemogenesis. PMID- 16721387 TI - Population genetics: DNAs from the European Neolithic. PMID- 16721388 TI - Among- and within-population variation in outcrossing rate of a mixed-mating freshwater snail. AB - Mixed-mating animals self-fertilize a proportion of their offspring. Outcrossing rate may covary with the ecological and historical factors affecting the population. Theory predicts that outcrossing is favored when inbreeding depression is high and when individual heterozygosity is important. Self fertilization is predicted to be favored when costs of male function, or mate finding are high, for example, when empty patches are colonized by few individuals. In this study, we assessed primary (after hatching) and secondary (after juvenile mortality) outcrossing rates of two mixed-mating snail populations. Our purpose was to assess the variation in mating-system parameters and estimate significance of inbreeding depression for secondary outcrossing rate (the realized outcrossing rate of parents that produce the next generation). Secondary outcrossing rate was higher than the primary outcrossing rate in one of the two populations, suggesting considerable inbreeding depression. In the other study population, secondary outcrossing rates were found to increase when initially low, or decrease when initially high, depending on the family. Moderate outcrossing rates were found to be more stable. Parental inbreeding coefficients were close to zero in both populations. Outcrossing rate was much more variable among families in the population with the lower average outcrossing rate, suggesting that individuals differed considerably in their mating system. Our results add to recent studies suggesting that populations of mixed-mating animals may differ in their mating system parameters and expression of inbreeding depression. PMID- 16721389 TI - Hox transcription factors and their elusive mammalian gene targets. AB - The Hox family of homeodomain transcription factors regulate numerous pathways during developmental and normal cellular processes. All Hox proteins recognise similar sequences in vitro yet display functional diversity in an in vivo environment. This review focuses on the transcriptional and functional specificity elicited by Hox proteins, giving an overview of homeodomain-DNA interactions and the gain of binding specificity through cooperative binding with cofactors. Furthermore, currently identified mammalian Hox target genes are presented, of which the most striking feature is that very few direct Hox targets have been identified. The direct targets participate in an array of cellular functions including organogenesis and cellular differentiation, cell adhesion and migration and cell cycle and apoptotic pathways. A further assessment of identified mammalian promoter targets and the contribution of bases outside the canonical recognition motif is given, highlighting roles they may play in either trans-activation or repression by Hox proteins. PMID- 16721390 TI - A novel mating system analysis for modes of self-oriented mating applied to diploid and polyploid arctic Easter daisies (Townsendia hookeri). AB - We have developed a new model for mating system analysis, which attempts to distinguish among alternative modes of self-oriented mating within populations. This model jointly estimates the rates of outcrossing, selfing, automixis and apomixis, through the use of information in the family structure given by dominant genetic marker data. The method is presented, its statistical properties evaluated, and is applied to three arctic Easter daisy populations, one consisting of diploids, the other two of tetraploids. The tetraploids are predominantly male sterile and reported to be apomictic while the diploids are male fertile. In each Easter daisy population, 10 maternal arrays of six progeny were assayed for amplified fragment length polymorphism markers. Estimates, confirmed with likelihood ratio tests of mating hypotheses, showed apomixis to be predominant in all populations (ca. 70%), but selfing or automixis was moderate (ca. 25%) in tetraploids. It was difficult to distinguish selfing from automixis, and simulations confirm that with even very large sample sizes, the estimates have a very strong negative statistical correlation, for example, they are not independent. No selfing or automixis was apparent in the diploid population, instead, moderate levels of outcrossing were detected (23%). Low but significant levels of outcrossing (2-4%) seemed to occur in the male-sterile tetraploid populations; this may be due to genotyping error of this level. Overall, this study shows apomixis can be partial, and provides evidence for higher levels of inbreeding in polyploids compared to diploids and for significant levels of apomixis in a diploid plant population. PMID- 16721391 TI - Genealogy of wine grape cultivars: "Pinot" is related to "Syrah". AB - Since the domestication of wild grapes ca 6000 years ago, numerous cultivars have been generated by spontaneous or deliberate crosses, and up to 10 000 are still in existence today. Just as in human paternity analysis, DNA typing can reveal unexpected parentage of grape cultivars. In this study, we have analysed 89 grape cultivars with 60 microsatellite markers in order to accurately calculate the identity-by-descent (IBD) and relatedness (r) coefficients among six putatively related cultivars from France ("Pinot", "Syrah" and "Dureza") and northern Italy ("Teroldego", "Lagrein" and "Marzemino"). Using a recently developed likelihood based approach to analyse kinship in grapes, we provide the first evidence of a genetic link between grapes across the Alps: "Dureza" and "Teroldego" turn out to be full-siblings (FS). For the first time in grapevine genetics we were able to detect FS without knowing one of the parents and identify unexpected second degree relatives. We reconstructed the most likely pedigree that revealed a third degree relationship between the worldwide-cultivated "Pinot" from Burgundy and "Syrah" from the Rhone Valley. Our finding was totally unsuspected by classical ampelography and it challenges the commonly assumed independent origins of these grape cultivars. Our results and this new approach in grape genetics will (a) help grape breeders to avoid choosing closely related varieties for new crosses, (b) provide pedigrees of cultivars in order to detect inheritance of disease resistance genes and (c) open the way for future discoveries of first- and second degree relationships between grape cultivars in order to better understand viticultural migrations. PMID- 16721392 TI - Paternal inheritance in parthenogenetic forms of the planarian Schmidtea polychroa. AB - Parthenogenesis usually includes clonal inheritance, which is thought to increase the risk of the clonal populations' extinction. Yet many parthenogenetic organisms appear to have survived for extended periods. A possible explanation is that parthenogens occasionally reproduce through sex-like processes. Although there is indirect evidence for occasional sex, the underlying mechanisms are currently unknown. In the present study, we examined sex-like processes in the planarian flatworm Schmidtea (Dugesia) polychroa. Parthenogenetic forms of this species are simultaneous hermaphrodites that require sperm to trigger embryogenesis, whereas paternal genetic material is usually excluded from the oocyte (sperm-dependent parthenogenesis). Based on a comparison of parents and offspring, using highly polymorphic microsatellites, we demonstrate the incorporation of paternal alleles in about 5% of the offspring. We detected two distinct processes: chromosome addition and chromosome displacement. Such rare sexual processes may explain the long-term persistence of the many purely parthenogenetic populations of S. polychroa in northern Europe. PMID- 16721393 TI - Do women give the same information on binge drinking during pregnancy when asked repeatedly? AB - OBJECTIVE: To study if pregnant women give the same answers to questions on frequency and timing of binge drinking when asked more than once during and after pregnancy. DESIGN: Cohort study. SETTING: The Danish National Birth Cohort. SUBJECTS: The study is based on 76 307 pregnant women with repeated information on binge drinking during the early part of pregnancy and 8933 pregnant women with information on binge drinking during pregnancy weeks 30-36, obtained while pregnant and 6 months after delivery. RESULTS: More women reported binge drinking, if the interview took place close to the period in question. As the report of binge drinking was highest in the first of two interviews referring to the same period, as well as women who participated in the first interview in pregnancy week 12 or earlier reported more binge drinking compared to women who participated in the interview later in pregnancy. CONCLUSIONS: Self-reported information on binge drinking is more frequently under-reported when the recall period is long. To improve the validity of data on binge drinking, future birth cohorts should obtain information several times during pregnancy. PMID- 16721394 TI - Dose-response study of probiotic bacteria Bifidobacterium animalis subsp lactis BB-12 and Lactobacillus paracasei subsp paracasei CRL-341 in healthy young adults. AB - OBJECTIVE: This study was performed to investigate the dose-response effects of supplementation with Bifidobacterium animalis subsp lactis (BB-12) and Lactobacillus paracasei subsp paracasei (CRL-431) on blood lipids, recovery from feces and bowel habits. Changes of the fecal microflora was analyzed in the 10(10) CFU/day probiotic and placebo group. DESIGN: The study was designed as a randomized, placebo-controlled, double-blinded, parallel dose-response study. SUBJECTS: Healthy young adults (18-40 years) were recruited by advertising in local newspapers. Of the 75 persons enrolled, 71 (46 women, 25 men, mean age 25.6 years (range 18-40 years)) completed the study. INTERVENTION: The volunteers were randomly assigned into five groups receiving either placebo or a mixture of the two probiotics in the concentration of 10(8), 10(9), 10(10) or 10(11) CFU/day in 2 weeks run-in period, 3 weeks intervention and 2 weeks wash-out. Diary reporting bowel habits and well being (abdominal bloating, flatulence and headache) was kept for all 7 weeks and blood lipids, fecal recovery of BB-12 and CRL-431, as well as fecal microflora was tested before, immediately and 2 weeks after intervention. RESULTS: The fecal recovery of BB-12 increased significantly (P < 0.001) with increasing dose. In the group receiving 10(11) CFU/day BB-12 was recovered from 13 out of 15 volunteers. CRL-431 was not recovered in any of the fecal samples. Supplementation with probiotics did not change the fecal bacterial composition. A significant linear increase in fecal consistency (looser stool) with increasing probiotic dose (P = 0.018) was observed. No overall dose-response effect was found on the blood lipids. High doses of probiotics were well tolerated. CONCLUSION: A dose-related recovery of BB-12 from feces was observed. PMID- 16721395 TI - Traditional food consumption and nutritional status of Dalit mothers in rural Andhra Pradesh, South India. AB - OBJECTIVES: To describe prevalence of malnutrition and their correlates of nutrient and traditional food consumption in rural Dalit mothers. DESIGN: In a cross-sectional study, we used socio-cultural questionnaires, anthropometric measurements and clinical eye examinations during the rainy season in 2003. Food frequency questionnaires and 24-h recalls were conducted during both summer and rainy seasons. SETTING: Dalit mothers with young children were recruited from 37 villages in the Medak District of rural Andhra Pradesh, India. SUBJECTS: Dalit mothers (n = 220) participated. RESULTS: The prevalence of chronic energy deficient (CED) mothers (body mass index <18.5 kg/m2) was 58%. Illiterate women and active women were more likely to have CED than those literate and non-active (relative risks (RR) = 1.6 and 1.4, respectively, P < or = 0.05), but literacy and activity level were not significant in multivariable analyses including sanitation and number of children < or =5 years of age. Increasing levels of fat intake, as a percent of total energy, was significantly associated with lower risk of CED (RR of the lowest 25th percentile compared to those in the 75th percentile or above was 1.6, P < or = 0.05), findings that remained significant in multivariable analyses. Consumption of pulses (g/day) was also inversely related to CED in univariate and multivariable analyses. Carbohydrate intake, as a percent of total energy, was inversely related to percent energy from fat (r = 0.96, P < or = 0.01), and, although positively related to CED in univariate analyses, carbohydrate consumption was not significant in multivariable analyses. Mothers' age in years and income was positively related to vitamin A deficiency. CONCLUSIONS: These results confirm that CED and vitamin A malnutrition among Dalit women are predominant problems in this area. Increased consumption of local traditional Dalit food (particularly sorghum, pulses, vegetables and animal source food) should be incorporated as an important component of intervention strategies to improve nutritional status. PMID- 16721396 TI - A community randomized controlled clinical trial of mixed carotenoids and micronutrient supplementation of patients with acquired immunodeficiency syndrome. AB - OBJECTIVE: This clinical trial aims to evaluate if natural mixed carotenoids supplementation can improve the health and survival of acquired immunodeficiency syndrome (AIDS) patients. DESIGN: A placebo-controlled, prospective, randomized, double-blind, multicenter clinical trial. SETTING: Community, tertiary care human immunodeficiency virus (HIV) clinics of the Canadian HIV Trials Network (CTN). PARTICIPANTS: Three hundred and thirty-one adults with advanced AIDS on conventional management were recruited during routine clinic visits. INTERVENTIONS: All participants, including 166 controls, received daily oral specially formulated multivitamins including vitamin A and trace elements; 165 treatment group participants received additional daily oral natural mixed carotenoids, equivalent to 120,000 IU (72 mg) of beta-carotene daily. Follow-up was quarterly at routine clinic visits. RESULTS: Mean (s.d.) follow-up was for 13 (6) months. Thirty-six participants died by 18 months. Serum carotene concentration <1.0 micromol/l was present in 16% participants at baseline. Despite variation in carotene content of the treatment medication, serum carotene concentrations increased significantly to twice the baseline levels to 18 months follow-up in participants who received carotenoids treatment compared with controls (P < 0.0001). Although not statistically significant, mortality was increased in participants who did not receive carotenoids treatment compared with those who did (HR time to death 1.76, 95% CI 0.89, 3.47, P = 0.11). In multivariate analysis, survival was significantly and independently improved in those with higher baseline serum carotene concentrations (P = 0.04) or higher baseline CD4 T-lymphocyte counts (P = 0.005). Adjusted mortality was also significantly and independently increased in those who did not receive carotenoids treatment compared with those who did (HR time to death 3.15, 95% CI 1.10, 8.98, P = 0.03). CONCLUSIONS: Low serum carotene concentration is common in AIDS patients and predicts death. Supplementation with micronutrients and natural mixed carotenoids may improve survival by correction of a micronutrient deficiency. Further studies are needed to corroborate findings and elucidate mechanism of action. PMID- 16721397 TI - Total energy expenditure, resting metabolic rate and physical activity level in free-living rural elderly men and women from Cuba, Chile and Mexico. AB - OBJECTIVE: The objective of this study was to assess total energy expenditure (TEE), resting metabolic rate (RMR) and physical activity level (PAL), and to estimate energy requirements (ERs) in free-living elderly subjects from Cuba, Chile and Mexico. DESIGN: Cross-sectional study designed to estimate ERs. SETTING: Rural regions of Cuba, Chile and Mexico. SUBJECTS: Forty subjects >60 years old were selected to participate in this study. INTERVENTIONS: A dose of doubly labeled water (DLW) was administered and urine samples were collected in the following 12-14 days. From the isotopic analysis, TEE was derived. RMR was measured by indirect calorimetry. RESULTS: TEE in Chilean (8.8+/-1.6 MJ/day) and Cuban (8.3+/-1.3 MJ/day) elderly was not different, and was higher for the Mexican group (9.5+/-1.5 MJ/day) (P < 0.0001). RMR was not different between countries. PAL and activity energy expenditure (AEE) were only different between Chile and Mexico (P < 0.002). For the three regions, overall PALs were 1.70+/ 0.16 for male and 1.62+/-0.13 for female subjects (P < 0.02), and AEE was 3.05+/ 0.66 and 2.27+/-0.66 MJ/day for male and female subjects, respectively (P < 0.001). Predictive equations (MJ/day) were RMR = 1.6447 + 0.05714 x weight (kg) + 0.449 sex (male = 1; female = 0) (R2 = 0.75; SEE = 0.479) and TEE = 3.414 + 0.0795 x weight (kg) + 1.227 x sex (male = 1; female = 0) (R2 = 0.75; SEE = 0.668). CONCLUSIONS: There were differences in TEE and PAL owing to sex and region. The average PAL in men was higher than the PAL reported either with factorial approach or with the DLW method in elderly. Predictive ER equations based on RMR and TEE gave very similar results to calculations from the 2004 FAO/WHO/UNU report. PMID- 16721398 TI - Effects of low-fat hard cheese enriched with plant stanol esters on serum lipids and apolipoprotein B in mildly hypercholesterolaemic subjects. AB - OBJECTIVE: To investigate the cholesterol-lowering effects of a low-fat cheese enriched with plant stanol esters in mildly hypercholesterolaemic subjects, as part of their normal diet. DESIGN: A randomized double-blind parallel-group study. SETTING: Valio Ltd, Helsinki. SUBJECTS: Sixty-seven mildly hypercholesterolaemic volunteers (24 men, 43 women) participated in the study, which all of them completed. INTERVENTIONS: The subjects were randomly assigned to the plant stanol ester group or the control group. During the 5-week intervention, the subjects in the stanol group consumed a cheese enriched with 2 g of plant stanols per day, and the subjects in the control group, a control cheese with no plant stanols. RESULTS: In the stanol ester group, as compared to the control group, both serum total and low-density lipoprotein (LDL) cholesterol decreased significantly, that is, by 5.8% (-0.32 mmol/l, 95% CI -0.50 to -0.15 mmol/l, P < 0.001) and 10.3% (-0.36 mmol/l, 95% CI -0.53 to -0.18 mmol/l, P < 0.001), respectively. There were no significant changes in high-density lipoprotein cholesterol (HDL), triglycerides or apolipoprotein B concentrations between the groups. CONCLUSION: Cheese enriched with 2 g of plant stanol in the form of fatty acid esters decreases serum total and LDL cholesterol significantly. PMID- 16721399 TI - Postpartum vitamin D insufficiency and secondary hyperparathyroidism in healthy Danish women. AB - OBJECTIVE: To examine vitamin D status and parathyroid function in normal Danish women postpartum. DESIGN: Three cross-sectional measures during follow-up of 89 women postpartum. SUBJECTS AND INTERVENTION: We assessed vitamin D status by measuring plasma 25-hydroxyvitamin D (P-25OHD) and the degree of secondary hyperparathyroidism by measuring plasma parathyroid hormone (P-PTH) in 89 Caucasian women at three consecutive visits: (mean (range)) 23 (10-37) days (spring), 117 (95-140) days (late summer) and 274 (254-323) days (winter) postpartum. RESULTS: P-25OHD showed seasonal variations with higher values in late summer than in the other periods (P < 0.001). At the first visit, 65% received vitamin D supplements. At the following visits, almost 50% were supplemented. Vitamin D insufficiency (P-25OHD < 50 nmol/l) occurred more often during winter (28%) than in spring (14%) (Fisher's exact test, P = 0.02) or late summer (7%) (P = 0.0001). Irrespective of season, vitamin D insufficiency occurred most frequent in women who did not take vitamin D supplements (Fisher's exact test, P < 0.02). Frank vitamin D deficiency (P-25OHD < 25 nmol/l) was observed during winter in 6%. At all three periods, P-25OHD correlated inversely with P-PTH indicating secondary hyperparathyroidism at deficient vitamin D status. During spring, late summer and winter three, one and four females, respectively, had elevated plasma PTH. CONCLUSION: Vitamin D insufficiency with secondary hyperparathyroidism is a frequent finding in healthy Danish women postpartum and especially during winter. Vitamin D supplements reduced the risk of vitamin D insufficiency, especially during winter. Our results support the importance of increased alertness regarding information of pregnant and lactating women about vitamin D supplements. Furthermore, it has to be studied whether the present recommendations of an intake of 5-10 microg vitamin D/day are sufficient, especially during winter months. PMID- 16721401 TI - The role of mutagenesis in defining genes in behaviour. AB - The study of human behavioural and psychiatric disorders benefits from the development of genetic models in mice and other organisms. Mouse mutants allow one to investigate the molecular basis of disease progression and to develop novel therapies. The number of potential mouse models is increasing dramatically through the implementation of mutagenesis screens for aberrant behavioural phenotypes. The alkylating agent N-ethyl-N-nitrosourea ENU is the mutagen of choice in these screens as it induces mutations at a very high rate. Progeny of chemically-mutagenised animals are screened either in systematic high-throughput test batteries or in specific low-throughput tests. Both approaches have been highly successful with large numbers of novel loci being identified and characterised. Many mutant lines are available for general research with phenotypes and genetic map positions on public websites. Of the mutant genes characterised, the majority have contributed to our knowledge of gene function in physiology and disease. The 'mutagenesis screening' approach continues to evolve through the design of new phenotyping strategies. The development of modifier screens in mice shows promise in the elucidation of complex phenotypes whereas the use of mutagenesis in combination with pharmacological agents targets specific neurochemical systems. Finally, the systematic screening approach has demonstrated that mutations are likely to affect more than one biological process. PMID- 16721402 TI - Genetics of affective (mood) disorders. AB - The enormous public health importance of mood disorders, when considered alongside their substantial heritabilities, has stimulated much work, predominantly in bipolar disorder but increasingly in unipolar depression, aimed at identifying susceptibility genes using both positional and functional molecular genetic approaches. Several regions of interest have emerged in linkage studies and, recently, evidence implicating specific genes has been reported; the best supported include BDNF and DAOA but further replications are required and phenotypic relationships and biological mechanisms need investigation. The complexity of psychiatric phenotypes is demonstrated by (a) the evidence accumulating for an overlap in genetic susceptibility across the traditional classification systems that divide disorders into schizophrenia and mood disorders, and (b) evidence suggestive of gene-environment interactions. PMID- 16721403 TI - Molecular genetic studies of schizophrenia. AB - The study of schizophrenia genetics has confirmed the importance of genes in etiology, but has not so far identified the relationship between observed genetic risks and specific DNA variants, protein alterations or biological processes. In spite of many limitations, numerous regions of the human genome give consistent, although by no means unanimous, support for linkage, which is unlikely to occur by chance. Two recent shifts have been evident in the field. First, a series of studies combining linkage and association analyses in the same family sets have identified promising candidate genes (DTNBP1, NRG1, G72/G30, TRAR4). Although a consensus definition of replication for genetic association in a complex trait remains difficult to achieve, the evidence for two of these (dystrobrevin binding protein 1 (DTNBP1), NRG1) is strong. Second, a series of studies combining association with functional investigation of changes in the associated gene in schizophrenia have also identified several candidate genes (COMT, RGS4, PPP3CC, ZDHHC8, AKT1). Somewhat surprisingly, the loci implicated by these studies have proven less robust in replication, although the number of replication studies remains small in several cases. Assessment of the combined evidence for the DTNBP1 gene gives some insight into the nature of the problems remaining to be solved. PMID- 16721404 TI - The genetics of developmental dyslexia. AB - Reading reflects the complex integration of several cognitive processes and proves more difficult to achieve for a significant proportion of the population. Developmental dyslexia (DD), or specific reading disability, is influenced by genes, a fact that has led several research groups to attempt to identify susceptibility genes through the sequential analysis of genetic linkage and association. Strong evidence has now emerged for the presence of genes influencing DD at several chromosomal loci and for at least one of these, there is evidence implicating specific genes. In this review, we present the evidence for a genetic contribution to DD and its component processes and review the current status of molecular genetic research aimed at identifying susceptibility genes for this common, complex disorder. PMID- 16721405 TI - Genetics of intelligence. AB - This article provides an overview of the biometric and molecular genetic studies of human psychometric intelligence. In the biometric research, special attention is given to the environmental and genetic contributions to specific and general cognitive ability differences, and how these differ from early childhood to old age. Special mention is also made of multivariate studies that examine the genetic correlation between intelligence test scores and their correlates such as processing speed, birth weight and brain size. After an overview of candidate gene associations with intelligence test scores, there is a discussion of whole genome linkage and association studies, the first of which have only recently appeared. PMID- 16721406 TI - Genetics and pathophysiology of mental retardation. AB - Mental retardation (MR) is defined as an overall intelligence quotient lower than 70, associated with functional deficit in adaptive behavior, such as daily-living skills, social skills and communication. Affecting 1-3% of the population and resulting from extraordinary heterogeneous environmental, chromosomal and monogenic causes, MR represents one of the most difficult challenges faced today by clinician and geneticists. Detailed analysis of the Online Mendelian Inheritance in Man database and literature searches revealed more than a thousand entries for MR, and more than 290 genes involved in clinical phenotypes or syndromes, metabolic or neurological disorders characterized by MR. We estimate that many more MR genes remain to be identified. The purpose of this review is to provide an overview on the remarkable progress achieved over the last decade in delineating genetic causes of MR, and to highlight the emerging biological and cellular processes and pathways underlying pathogeneses of human cognitive disorders. PMID- 16721407 TI - Genetics of autism spectrum disorder. AB - Autism is a highly heritable complex neurodevelopmental disorder characterized by distinct impairments of cognitive function in the field of social interaction and speech development. Different approaches have been undertaken worldwide to identify susceptibility loci or genes for autism spectrum disorders. No clear conclusions can be made today about genetic loci involved in these disorders. The review will focus on relevant results from the last decade of research with emphasis on whole genome screens and association studies. PMID- 16721408 TI - The genetic basis of emotional behaviour in mice. AB - The last decade has witnessed a steady expansion in the number of quantitative trait loci (QTL) mapped for complex phenotypes. However, despite this proliferation, the number of successfully cloned QTL has remained surprisingly low, and to a great extent limited to large effect loci. In this review, we follow the progress of one complex trait locus; a low magnitude moderator of murine emotionality identified some 10 years ago in a simple two-strain intercross, and successively resolved using a variety of crosses and fear-related phenotypes. These experiments have revealed a complex underlying genetic architecture, whereby genetic effects fractionate into several separable QTL with some evidence of phenotype specificity. Ultimately, we describe a method of assessing gene candidacy, and show that given sufficient access to genetic diversity and recombination, progression from QTL to gene can be achieved even for low magnitude genetic effects. PMID- 16721409 TI - Molecular genetics of the fruit-fly circadian clock. AB - The circadian clock percolates through every aspect of behaviour and physiology, and has wide implications for human and animal health. The molecular basis of the Drosophila circadian clock provides a model system that has remarkable similarities to that of mammals. The various cardinal clock molecules in the fly are outlined, and compared to those of their actual and 'functional' homologues in the mammal. We also focus on the evolutionary tinkering of these clock genes and compare and contrast the neuronal basis for behavioural rhythms between the two phyla. PMID- 16721410 TI - Utility of urinary 1-naphthol and 2-naphthol levels to assess environmental carbaryl and naphthalene exposure in an epidemiology study. AB - We recently reported associations between urinary 1-naphthol (1N) levels and several intermediate measures of male reproductive health, namely sperm motility, serum testosterone levels, and sperm DNA damage. However, because 1N is a major urinary metabolite of both naphthalene and the insecticide carbaryl, exposure misclassification stemming from differences in exposure source was probable and interpretation of the results was limited. As naphthalene, but not carbaryl, is also metabolized to 2-naphthol (2N), the relationship of urinary 1N to 2N within an individual may give information about source of 1N. Utilizing data from two previous studies that measured both 1N and 2N in urine of men exposed to either carbaryl or naphthalene, the present study employed several methods to differentiate urinary 1N arising from exposures to carbaryl and naphthalene among men in the reproductive health study. When re-evaluating the reproductive health data, techniques for identifying 1N source involved exploring interaction terms, stratifying the data set based on 1N/2N ratios, and performing an exposure calibration using a linear 1N to 2N relationship from a study of workers exposed to naphthalene in jet fuel. Despite some inconsistencies between the methods used to distinguish 1N source, we found that 1N from carbaryl exposure is likely responsible for the previously observed association between 1N and sperm motility, whereas 1N from naphthalene exposure is likely accountable for the association between 1N and sperm DNA damage. We demonstrate that studies of health effects associated with carbaryl should utilize a 1N/2N ratio to identify subgroups in which carbaryl is the primary source of 1N. Conversely, studies of naphthalene-related outcomes may utilize 2N levels to estimate exposure. PMID- 16721411 TI - The impact of improved wood-burning stoves on fine particulate matter concentrations in rural Mexican homes. AB - To evaluate the impact of improved wood burning stoves on indoor air pollution, 53 homes in a rural town in Michoacan, Mexico, were selected from a health intervention study and monitored before and after receiving improved wood-burning stoves. Fine particulate matter--particles with aerodynamic diameter less than 2.5 microm (PM(2.5))--concentrations were measured in the central plaza of the community and in three microenvironments in the home (next to the stove, in the kitchen away from the stove, and outdoor patio). Forty-eight hour mean PM(2.5) concentrations in homes that burned wood in open fires were 693 microg/m(3) (95% CI: 246-1338) near the stove, 658 microg/m(3) (95% CI: 67-1448) in the kitchen away from the stove, and 94 microg/m(3) (95% CI: 36-236) on the patio. Mean ambient 24-h concentrations in the main plaza of the community were 59 microg/m(3) (95% CI: 29-92). Paired measurements before and after the installation of the Patsari improved wood-burning stove indicate a median 71% reduction in PM(2.5) concentrations near the stove and 58% reductions in kitchen concentrations, whereas patio and main plaza concentrations remain unaffected. Only 44% of participants reported to use their Patsari stoves exclusively during the transition period. Even with the predominant mixed use of the Patsari stove with open fires, estimated daily average personal exposures to PM(2.5) were reduced by 50%. PMID- 16721412 TI - Real-time measurement of short-term peaks in environmental CO concentrations in the homes of the elderly in South Wales. AB - In order to determine whether short-term World Health Organization (WHO) guideline limits for carbon monoxide (CO) are exceeded in the home environment, a number of sensors were constructed capable of real-time measurement. These were deployed for a period of 7 days to continuously monitor CO concentrations in 44 non-smoking households using either gas, coal, electricity, liquid propane gas, or oil for heating fuel in South Wales. The mean environmental concentrations over the period monitored were less than 1 p.p.m and WHO short-term limits were not exceeded in any household. In another 20 homes containing a smoker the mean concentrations measured over a period of 2 days ranged from 0.1 to 21 p.p.m and WHO short-term limits were exceeded in one household. PMID- 16721413 TI - Indoor time-microenvironment-activity patterns in seven regions of Europe. AB - Personal exposure to environmental substances is largely determined by time microenvironment-activity patterns while moving across locations or microenvironments. Therefore, time-microenvironment-activity data are particularly useful in modeling exposure. We investigated determinants of workday time-microenvironment-activity patterns of the adult urban population in seven European cities. The EXPOLIS study assessed workday time-microenvironment activity patterns among a total of 1427 subjects (age 19-60 years) in Helsinki (Finland), Athens (Greece), Basel (Switzerland), Grenoble (France), Milan (Italy), Prague (Czech Republic), and Oxford (UK). Subjects completed time microenvironment-activity diaries during two working days. We present time spent indoors--at home, at work, and elsewhere, and time exposed to tobacco smoke indoors for all cities. The contribution of sociodemographic factors has been assessed using regression models. More than 90% of the variance in indoor time microenvironment-activity patterns originated from differences between and within subjects rather than between cities. The most common factors that were associated with indoor time-microenvironment-activity patterns, with similar contributions in all cities, were the specific work status, employment status, whether the participants were living alone, and whether the participants had children at home. Gender and season were associated with indoor time-microenvironment activity patterns as well but the effects were rather heterogeneous across the seven cities. Exposure to second-hand tobacco smoke differed substantially across these cities. The heterogeneity of these factors across cities may reflect city specific characteristics but selection biases in the sampled local populations may also explain part of the findings. Determinants of time-microenvironment activity patterns need to be taken into account in exposure assessment, epidemiological analyses, exposure simulations, as well as in the development of preventive strategies that focus on time-microenvironment-activity patterns that ultimately determine exposures. PMID- 16721414 TI - [The immigrant in primary care consultations]. AB - In consultations at health centres, the GP, the paediatrician and the nursing staff have faced, above all since the end of the XIX century and so far in the XXI century, the fact of having to attend to a numerous population formed of people uprooted from their community, without close relatives in the majority of cases, with different languages and cultures, and with a different way of understanding health and illness. This article analyses this phenomenon and aims to improve the understanding of health professionals and contribute to improving care for the immigrant patient. PMID- 16721415 TI - [Medical emergencies in the adult immigrant]. AB - The need for medical care of the immigrant population is growing in proportion to the increase in the number of immigrants resident in Spain. This article reviews the socio-demographic characteristics of the immigrant population that come for consultations in the emergency services and discusses those particularities, both medical (neurocysticercosis, tuberculosis, infection by the AIDS virus, hepatitis, malaria, parasitosis, Ulysses syndrome) and paramedical (knowledge of the health card, concept of medicine, language barrier), that differentiate them from the native population. Nonetheless, since we are dealing with a young population that is in principle healthy, their reasons for consultation do not greatly differ from the general population. It is worth drawing attention to the high level of satisfaction of this population with the care afforded them in the hospital emergency services. PMID- 16721416 TI - [Assessment of the immigrant child]. AB - In the last twenty years Spain has turned into one of the principal countries recipients of foreign population. As consequence of this massive entry of people, our country is among the first ones of Europe for interannual increase of population. The concept of immigrant child is very wide and includes minors of very diverse characteristics, as the proceeding from international adoptions, the children coming from developing countries or those that come from other developed countries. The immigrant children do not represent health risk for the autochthonous population, on the contrary, they are in situation of defenselessness for their scanty vaccination coverages, their situation of marginality and the precarious sanitary systems of the countries of origin. At the moment of offering medical paediatric attention to foreign children, we must individualize our actions in conformity with the characteristics of each patient. It is not possible to simplify in one unique protocol the best studies to resolve a problem of health in an immigrant child. The attention of these children implies an overstrain for the pediatrician and the sanitary personnel that receives them, because it is necessary to conquer idiomatic, cultural and social barriers to optimize the level of health of these patients. The syndromic initial description can help to prioritize the studies in each case (according to the most probable diagnoses). The children who return to the native land of their parents meet in a special situation of risk, for not being prepared to resist the attack of pathogen that do not exist or are rare in Spain. PMID- 16721417 TI - [Antenatal care in immigrants]. AB - The phenomenon of immigration has had an impact on the health care of the population. The immigrant population in Spain today represents approximately 8% of the total population. The majority of this population proceeds from countries with low income, and its origin and distribution is diverse. The immigrant population is characterised by its being young and healthy, and with a capacity to adapt to changes, but its social, economic and labour conditions are frequently insecure and favour vulnerability to disease. In spite of the number of immigrants of the male sex being globally higher than that of women, the percentage of immigrants of the female sex is growing. This increase of the female immigrant population has resulted in the appearance of specific health care needs, especially with respect to sexual and reproductive health. To which we must add a substantial increase in pathologies prevalent in the countries of origin, such as anaemia, tuberculosis, malnutrition, haemoglobinopathies, consanguinity, hypocalcaemia, hepatitis B and/or C, sexually transmitted infections, infectious diseases transmitted by arthropods, such as Chagas disease and other parasitoses, as well as genital mutilations. The aim of this article is to analyse the factors that make it difficult to control gestation in the immigrant population, as well as to establish guidelines for acting in antenatal care consultations. Insistence is placed on health education and prevention during pregnancy, and consideration is given to the appearance of rare diseases related to some of these groups. PMID- 16721418 TI - [Migration and psychiatric diseases]. AB - Immigration is an emergent social phenomenon with a great impact on health systems. Psychiatric disorders are said to be universal phenomenon, but their clinical expression may be determined by cultural factors. Indeed, immigration acts as a risk factor for the development of mental diseases, as it works as a stress generating factor. The adaptability process for both the immigrants and the receptor environment, involves a social and cultural effort that modifies interindividual relationships, mainly at the therapeutic level. Psychiatry faces up to a new therapeutic and diagnostic challenge, where ethnocultural barriers (either individual or collective) and social and biological barriers need to be got over. Barriers to access to Mental Health Services or social resources and language differences mean an important obstacle which has to be got over by health systems providing special mental health programs, either segregationist or integrative. This review tries to summarize the different issues that limit or interfere with daily clinical practice when treating the immigrant population, meaningful both in quantitative and qualitative psychiatric terms. PMID- 16721419 TI - [Labour market, occupational health and immigration]. AB - The process of economic and social change that Navarra has undergone in recent decades has been associated with the arrival of a growing flow of immigrants since the start of the new century. They have had a decisive influence as a factor of economic change in terms of production increase and internal demand. A new Navarra is being built thanks to the phenomenon of migration. In the first place, we analyse their impact on demographic growth. Their influence on the labour market, with its highlights and shadows, is evaluated. Foreigners already are about 10% of the active population and their presence in some productive sectors is decisive for their viability. The dysfunctions and problems of the labour market are reviewed, especially the question of accidents. Finally, the behaviour of the indicators of temporary disability of this collective are set out. Although this phenomenon has brought an imbalance in some spheres of social policy (education, housing, health), it can be said that the model of integration in Navarra is based on a generous welfare system, a social climate that is in general tolerant, and sustained economic growth. PMID- 16721420 TI - [Tropical medicine consultation]. AB - Tropical medicine consultations are fully justified in settings with the latest modern technology, where specific complementary tests are available and there are professionals with experience in tropical questions. That is to say, in tertiary hospitals. If such consultations took place in secondary hospitals or in primary care, they could be considered inefficient or unjustifiable from the point of view of the volume of patients attended to. However, there is a care deficit with respect to preventive activities concerning travellers or immigrants who have recently arrived from countries with a low income and where there is a high prevalence of imported diseases that are less recognised in our normal health milieu. Thus, international health units, which combine preventive and curative activities in a framework of public health provision and in a functional situation between the hospital level and that of primary care, offer a more efficient and suitable profile for the characteristics of the Spanish population. Their implementation depends on policy makers, the offer of a realistic portfolio of services, the existence of quality control monitoring and the possibility of managing information through a computer network. PMID- 16721421 TI - [Prevention of diseases in the traveller; the returning traveller]. AB - In recent years there has been a spectacular increase in international journeys and the WHO calculates that by the year 2015 the number of long distance journeys will have doubled in comparison with twenty years before. Travelling involves a series of risks, which become higher the poorer the social and health situation of the country of destination. According to different studies, between 20 and 70% of travellers suffer from some type of health problem during their journey. The main causes of mortality during a journey are, in order of importance, accidents, cardiovascular diseases and transmissible diseases. This article sets out the current criteria and recommendations regarding general health advice, recommended vaccinations, anti-malaria chemoprophylaxis and special physiological (children and the pregnant women) and clinical (chronic diseases) situations of interest to travellers, in order to prevent health problems and as far as possible to guarantee a safe and profitable journey. PMID- 16721422 TI - [Malaria imported by immigrants]. AB - The immigrants from endemic zones who live in Europe are those with the greatest risk of acquiring malaria during trips to their countries, since they usually travel for longer than other travellers and to zones and under conditions of greater risk. The figures for the years 2002-2004 in Spain indicate that P. falciparum is the most diagnosed species. Africa is the continent from which most cases are imported and immigrants contribute 34-41% of all cases. PMID- 16721423 TI - [Diarrhoea in the traveller]. AB - Diarrhoeic diseases caused by water and food contaminated by enteropathogens continue to be an important cause of morbidity in countries with a low level of development. Some 50,000 cases of diarrhoea in travellers are estimated in the world every day, and this is the main cause of consultation by travellers who return from undeveloped zones. The principal determinant of risk is the place of destination; there are significant differences between different regions with respect to risk and to the aetiology of the diarrhoea. The most frequent cause of diarrhoeas is of bacterial origin, which represents between 60 and 85% of the cases, while parasites represent 10% and some 5% are produced by viruses. Although it normally follows a benign course, complications can arise, with mortality being only exceptionally associated to this disease. Prevention is essentially based on strictly following elemental hygienic measures and avoiding the ingestion of foodstuffs and drinks with a risk of contamination. Prophylaxis with antibiotics is only advisable in journeys of short duration, in which the risk and/or seriousness of diarrhoeas, above all in immunosuppressed patients, are higher than the possible collateral effects. The treatment of diarrhoea in the traveller is based on adequate hydration, and the use of microbians is reserved for moderate and serious situations, with quinolones being the drug of choice. Rifaximine is a new drug approved for the treatment of diarrhoeas in the traveller, above all in areas with enteropathogens that are resistant to quinolones. PMID- 16721424 TI - [Imported diseases in immigrants: myth and reality]. AB - In recent years, with the growth of immigration from low income countries, in certain health spheres and in civil society itself through the mass media, there has been speculation about the risk to the health of the receptor community brought by immigration. This article attempts to conceptualise the terminology that is used to deal with the diseases of immigrants, to serve as a common denominator in the description and interpretation of the data provided by the medical journals, avoiding the confusions that arise in non-specialised health settings or in those that are remote from this. It aims to reflect on the reality of these diseases, both those that are imported and those that are transmissible, and to clarify the risks for the receptor community and their determinants. PMID- 16721425 TI - Cryomacroscopy of vitrification, Part I: A prototype and experimental observations on the cocktails VS55 and DP6. AB - A new imaging device, termed a "cryomacroscope", is presented in this report. This device is designed to assist in exploring thermal and mechanical effects associated with large-scale vitrification and crystallization, with the current setup aimed at the range of 50 MUm to 2 cm. The cryomacroscope is not intended as a substitute for the cryomicroscope, but as a complementary tool for the cryobiologist. A combination of cryomacroscopy and cryomicroscopy is suggested as a basis for multi-scale cryobiology studies. This report presents initial results on vitrification, crystallization, and fracture formation in the cryoprotectant cocktails DP6 and VS55. These results show some inconsistency in the tendency to form crystals, based on critical cooling and rewarming rates measured by means of a differential scanning calorimetric device (DSC) in parallel studies. This research is in its early stages, and comparative studies on biological materials are currently underway. Part II of this report (the companion paper) presents results for fracture formation in the cryoprotectant and discusses the mechanical stresses which promote these fractures. In conjunction with these reports, additional photos of cryomacroscopy of vitrification, crystallization, and fracture formation are available at http://www.me.cmu.edu/faculty1/rabin/CryomacroscopyImages01.htm. PMID- 16721427 TI - Physico-chemical processes in imidazolium ionic liquids. AB - Among the various properties exhibited by ionic liquids (ILs)--especially those based on the imidazolium cation-their inherent ionic patterns, very low vapour pressure and pronounced self-organization in the solid, liquid and even in the gas phase are particularly interesting since this allows the use of these fluids as alternative and complementary media to classical organic solvents and water in many applications. Hence, reaction paths that involve charge-separated intermediates or transition states are accelerated--by lowering the activation barrier-in the presence of ILs when compared with those performed in classical organic solvents. It is also possible, for example, to observe, by electrochemical methods, transient species (ionic and radical) that are otherwise undetectible in water or in molecular organic solvents and to investigate the interactions and behaviour of molecular, biological and macromolecular species in solution using physical and chemical methods which require special conditions such as high-vacuum, and which have been traditionally limited to solid state chemistry. PMID- 16721426 TI - Identification of novel cell-adhesion molecules in peripheral nerves using a signal-sequence trap. AB - The development and maintenance of myelinated nerves in the PNS requires constant and reciprocal communication between Schwann cells and their associated axons. However, little is known about the nature of the cell-surface molecules that mediate axon-glial interactions at the onset of myelination and during maintenance of the myelin sheath in the adult. Based on the rationale that such molecules contain a signal sequence in order to be presented on the cell surface, we have employed a eukaryotic-based, signal-sequence-trap approach to identify novel secreted and membrane-bound molecules that are expressed in myelinating and non-myelinating Schwann cells. Using cDNA libraries derived from dbcAMP stimulated primary Schwann cells and 3-day-old rat sciatic nerve mRNAs, we generated an extensive list of novel molecules expressed in myelinating nerves in the PNS. Many of the identified proteins are cell-adhesion molecules (CAMs) and extracellular matrix (ECM) components, most of which have not been described previously in Schwann cells. In addition, we have identified several signaling receptors, growth and differentiation factors, ecto-enzymes and proteins that are associated with the endoplasmic reticulum and the Golgi network. We further examined the expression of several of the novel molecules in Schwann cells in culture and in rat sciatic nerve by primer-specific, real-time PCR and in situ hybridization. Our results indicate that myelinating Schwann cells express a battery of novel CAMs that might mediate their interactions with the underlying axons. PMID- 16721428 TI - On the fraction of CrII sites involved in the C2H4 polymerization on the Cr/SiO2 Phillips catalyst: a quantification by FTIR spectroscopy. AB - An estimation of the fraction of Cr(II) sites involved in the C(2)H(4) polymerization on a Cr(II)/SiO(2) Phillips catalyst has been obtained by means of in situ alternated CO adsorption and C(2)H(4) polymerization FTIR experiments: about 28% of the total surface sites react fast with C(2)H(4), while a lower fraction, which depends upon the temperature reaction conditions, is more slowly involved, in agreement with XANES results. PMID- 16721429 TI - Protons solvated in noble-gas matrices: interaction with nitrogen. AB - The interaction of a (NgHNg)(+) cation (Ng = Ar and Kr) with a nitrogen molecule is studied. The structure, energetics, and vibrational properties of these complexed systems are computationally studied at the MP2/6-311++G(2d,2p) level of theory. The computations reveal two stable structures, linear and T-shaped configurations, with BSSE corrected interaction energies of the order of -1000 cm(-1). The (NgHNg)(+)[dot dot dot]N(2) complexes are characterized experimentally by IR absorption spectroscopy in solid Ar and Kr matrices. The spectra show that only one complex structure is present, as evidenced by the single nitrogen-induced nu(3) band. According to the computational results, the linear structure is more probable in the experiments. However, our results show that the one-to-one complex at the present computational level does not accurately agree with the matrix-isolation experiments, the differences originating possibly from the influence of the surrounding matrix. Based on the current data, the mechanism of cation decay in noble-gas matrices is discussed. The observed similar decay of (NgHNg)(+) and its N(2) complex indicates that the solvated proton is unable to tunnel and is therefore immobile in noble-gas matrices. The observations for the cation decay are consistent with the electron neutralization mechanism. PMID- 16721430 TI - Periodic ab initio study of structural and vibrational features of hexagonal hydroxyapatite Ca10(PO4)6(OH)2. AB - Structural and vibrational features of hexagonal hydroxyapatite HA [Ca(10)(PO(4))(6)(OH)(2), space group P6(3)] are computed ab initio within a periodic approach using the CRYSTAL03 program and the B3LYP hybrid functional with a Gaussian-type basis set of polarized double zeta quality. Experimental lattice parameters and internal coordinates have been fully optimized and the final structure characterized by means of its band structure, density of states and Mulliken analysis. The full B3LYP harmonic vibrational spectrum of HA at Gamma point has also been computed and compares well with the available experimental IR and Raman data. Nevertheless, the presence of one negative frequency in the computed spectrum shows that, within the hexagonal symmetry imposed by the P6(3) group, the structure is a saddle point. This is at variance with the monoclinic structure (under P2(1)/b space group), which has been computed, with the same approach, to be a minimum of 17 kJ mol(-1) (per unit cell) more stable than the corresponding hexagonal HA structure. PMID- 16721431 TI - Lineshapes in carbon 1s photoelectron spectra of methanol clusters. AB - A general protocol for theoretical modeling of inner-shell photoelectron spectra of molecular clusters is presented and applied to C1s spectra of oligomers and medium-sized clusters of methanol. The protocol employs molecular dynamics for obtaining cluster geometries and a polarizable force field for computing site specific chemical shifts in ionization energy and linewidth. Comparisons to spectra computed from first-principle theories are used to establish the accuracy of the proposed force field approach. The model is used to analyze the C1s photoelectron spectrum of medium-sized clusters in terms of surface and bulk contributions. By treating the surface-to-bulk ratio as an adjustable parameter, satisfactory fits are obtained to experimental C1s spectra of a beam of methanol clusters. PMID- 16721432 TI - Experimental infrared spectra of Cl-(ROH) (R = H, CH3, CH3CH2) complexes in the gas-phase. AB - Infrared multiple photon dissociation spectra for the chloride ion solvated by either water, methanol or ethanol have been recorded using an FTICR spectrometer coupled to a free-electron laser, and are presented here along with assignments to the observed bands. The assignments made to the Cl(-)/H(2)O, Cl(-)(CH(3)OH), and Cl(-)(CH(3)CH(2)OH) spectra are based on comparison with the neutral H(2)O, CH(3)OH, and CH(3)CH(2)OH spectra, respectively. This work confirms that a band observed around 1400 cm(-1) in the Cl(-)(H(2)O) spectrum is not due to the Ar tag in Ar predissociation spectra. The carrier of this band is, most likely, the first overtone of the OHCl bend. Based on the position of the overtone in the IRMPD spectrum, 1375 cm(-1), the fundamental must occur very close to 700 cm(-1) and observation of this band should aid theoretical treatments of the spectrum of this complex. B3LYP/6-311++G(2df,2pd) calculations are shown to reproduce the IRMPD spectra of all three solvated chloride species. They also predict that attaching one or two Ar atoms to the Cl(-)(H(2)O) complex results in a shift of no more than a few wavenumbers in the fundamental bands for the bare complex, in agreement with previous experiment. For both alcohol-Cl(-) complexes, the S(N)2 "backside attack" isomers are not observed and Cl(-) is predicted theoretically, and confirmed experimentally, to be bound to the hydroxyl hydrogen. For Cl( )(CH(3)CH(2)OH), the trans and gauche conformers are similar in energy, with the gauche conformer predicted to be thermodynamically favoured. The experimental infrared spectrum agrees well with that predicted for the gauche conformer but a mixture of gauche and anti conformers cannot be ruled out based on the experimental spectra nor on the computed thermochemistry. PMID- 16721433 TI - Jet-cooled diode laser spectra of CF3Br in the 9.2 microm region and rovibrational analysis of symmetric CF3 stretching mode. AB - Pulsed slit-jet high resolution (up to 0.0009 cm(-1) FWHM) infrared diode laser spectra of CF(3)Br, with natural isotopic abundance, were obtained in the region around 9.2 microm at the rotational temperature of about 50 K. In addition, diode laser spectra at reduced temperature (200 K) were recorded. We present here the rovibrational analysis of the nu(1) fundamental in the range 1075-1090 cm(-1). The fine structure of many P(J) and R(J) clusters has been well resolved for the first time. The assignment of rovibrational transitions has been accomplished up to K = 27, J = 63 for CFBr and K = 33, J = 62 for CFBr. A total of 636 (CFBr) and 880 (CFBr) lines were used in the final fit and a very accurate set of molecular constants, including the quartic centrifugal distortion coefficients for the v(1) = 1 state of both the bromine isotopologues, was obtained. In addition, spectral features belonging to the nu(1) + nu(6)-nu(6) hot band were unambiguously identified and a set of spectroscopic parameters were determined. PMID- 16721434 TI - Extremely strong solvent dependence of the S1--> S0 internal conversion lifetime of 12'-apo-beta-caroten-12'-al. AB - The ultrafast internal conversion (IC) dynamics of the carbonyl carotenoid 12' apo-beta-caroten-12'-al has been investigated in solvents of varying polarity using time-resolved femtosecond transient absorption spectroscopy. The molecules were excited to the S(2) state by a pump beam of either 390 or 470 nm. The subsequent intramolecular dynamics were detected at several probe wavelengths covering the S(0)--> S(2) and S(1)--> S(n) bands. For the S(1)--> S(0) internal conversion process, a remarkably strong acceleration with increasing polarity was found, e.g., lifetimes of tau(1) = 220 ps (n-hexane), 91 ps (tetrahydrofuran) and 8.0 ps (methanol) after excitation at 390 nm. The observation can be rationalized by the formation of a combined S(1)/ICT (intramolecular charge transfer) state in the more polar solvents. The effect is even stronger than the strongest one reported so far in the literature for peridinin. Addition of lithium salts to a solution of 12'-apo-beta-caroten-12'-al in ethanol leads only to small changes of the IC time constant tau(1). In addition, we estimate an upper limit for the time constant tau(2) of the S(2)--> S(1) internal conversion process of 300 fs in all solvents. PMID- 16721435 TI - Ozonolysis and photolysis of alkene-terminated self-assembled monolayers on quartz nanoparticles: implications for photochemical aging of organic aerosol particles. AB - Photolysis of alkene-terminated self assembled monolayers (SAM) deposited on Degussa SiO(2) nanoparticles is studied following oxidation of SAM with a gaseous ozone/oxygen mixture. Infrared cavity ring-down spectroscopy is used to observe gas-phase products generated during ozonolysis and subsequent photolysis of SAM in real time. Reactions taking place during ozonolysis transform alkene terminated SAM into a photochemically active state capable of photolysis in the tropospheric actinic window (lambda > 295 nm). Formaldehyde and formic acid are the observed photolysis products. Photodissociation action spectra of oxidized SAM and the observed pattern of gas-phase products are consistent with the well established Criegee mechanism of ozonolysis of terminal alkenes. There is strong evidence for the presence of secondary ozonides (1,3,4-trioxalones) and other peroxides on the oxidized SAM surface. The data imply that photolysis plays a role in atmospheric aging of primary and secondary organic aerosol particles. PMID- 16721436 TI - Electrocatalytic oxidation of ammonia on Pt(111) and Pt(100) surfaces. AB - The electrocatalytic oxidation of ammonia on Pt(111) and Pt(100) has been studied using voltammetry, chronoamperometry, and in situ infrared spectroscopy. The oxidative adsorption of ammonia results in the formation of NH(x) (x = 0-2) adsorbates. On Pt(111), ammonia oxidation occurs in the double-layer region and results in the formation of NH and, possibly, N adsorbates. The experimental current transients show a hyperbolic decay (t(-1)), which indicates strong lateral (repulsive) interactions between the (reacting) species. On Pt(100), the NH(2) adsorbed species is the stable intermediate of ammonia oxidation. Stabilization of the NH and NH(2) fragments on Pt(111) and Pt(100), respectively, is in an interesting agreement with recent theoretical predictions. The Pt(111) surface shows extremely low activity in ammonia oxidation to dinitrogen, thus indicating that neither NH nor N (strongly) adsorbed species are active in dinitrogen production. Neither nitrous oxide nor nitric oxide is the product of ammonia oxidation on Pt(111) at potentials up to 0.9 V, as deduced from the in situ infrared spectroscopy measurements. The Pt(100) surface is highly active in dinitrogen production. This process is characterized by a Tafel slope of 30 mV decade(-1), which is explained by a rate-determining dimerization of NH(2) fragments followed by a fast decay of the resulting surface-bound hydrazine to dinitrogen. Therefore, the high activity of the Pt(100) surface for ammonia oxidation to dinitrogen is likely to be related to its ability to stabilize the NH(2) adsorbate. PMID- 16721437 TI - Particle-surface capillary forces with disjoining pressure. AB - A new, atomic force microscopy (AFM) based experimental setup for the continuous acquisition of friction force data as a function of humidity has been developed. The current model of interactions between wet contacts under the influence of capillary effects, has been amended to include a vertical component due to the disjoining pressure and takes into account the influence of liquid films adsorbed on the surface. This is a 'switching' model, i.e. the contact between nanometer sized sphere and a flat surface can exist in two distinct states due to capillary bridge formation/destruction as the humidity is varied. The model has been qualitatively verified on samples of differing wettability produced by UV-ozone treatment of polystyrene (PS). Results of AFM analysis of the friction vs. vapor pressure curves collected from the surface are presented. Correlation between important surface properties such as wettability, adsorption, and contact angle and friction force under varying humidity were found and discussed. PMID- 16721438 TI - Probing the validity of the Derjaguin approximation for heterogeneous colloidal particles. AB - The Derjaguin approximation states that the interaction force between two curved surfaces is proportional to their effective radius, whereby the inverse effective radius is the arithmetic mean of the inverse curvature radii of the surfaces involved. The present study investigates the validity of this approximation with an atomic force microscope (AFM) by measuring interaction forces between colloidal particles of different sizes, but of identical composition. Forces were measured between silica particles of 2.0, 4.8 and 6.8 microm in diameter in KCl electrolyte solution with and without adsorbed poly(amido amine) (PAMAM) dendrimers. The Derjaguin approximation could be confirmed at all distances investigated, including those comparable with the characteristic length scales of the surface roughness or the surface charge heterogeneities. For the conditions investigated, the Derjaguin approximation turns out to be surprisingly robust. PMID- 16721439 TI - Nano-jewellery: C5Au12--a gold-plated diamond at molecular level. AB - A mixed carbon-metal cluster is designed by combining the tetrahedral C(5) radical (with a central atom-the skeleton of the C(5)H(12) molecule) and the spherical Au(12) layer (the external atomic shell of the Au(13) cluster). The C(5)Au(12) cluster and its negative and positive ionic derivatives, C(5)Au(12)(+/ ), are investigated ab initio (DFT) in terms of optimized structures and relative energies of a few spin-states, for the icosahedral-like and octahedral-like isomers. The cluster is predicted to be generally more stable in its octahedral shape (similar to C(5)H(12)) which prevails for the negative ion and may compete with the icosahedral shape for the neutral system and positive ion. Adiabatic ionization energies (AIE) and electron affinities (AEA) of C(5)Au(12), vertical electron-detachment (VDE) energies of C(5)Au(12)(-), and vertical ionization and electron-attachment energies (VIE, VEA) of C(5)Au(12) are calculated as well, and compared with those for the corresponding isomers of the Au(13) cluster. The AIE and VIE values are found to be close for the two systems, while the AEA and VDE values are significantly reduced for the radical-based species. A simple fragment based model is proposed for the decomposition of the total interaction into carbon-gold and gold-gold components. PMID- 16721440 TI - Impact of pneumococcal conjugate vaccine on the prevention of invasive pneumococcal diseases. AB - OBJECTIVES: To evaluate the impact of heptavalent pneumococcal conjugate vaccine in invasive pneumococcal diseases in the United States, and to analyze the potential impact of this vaccine in Brazil. SOURCES OF DATA: MEDLINE, LILACS, Cochrane Database Reviews, as well as the websites of the Centers for Disease Control and Prevention (CDC), Brazilian Ministry of Health and Centro de Vigilancia Epidemiologica do Estado de Sao Paulo from January 2000 to January 2006. Articles retrieved with the keywords Streptococcus pneumoniae, pneumococcal diseases, conjugate vaccine, antimicrobial resistance and meningitis were reviewed. SUMMARY OF THE FINDINGS: The introduction of heptavalent pneumococcal conjugate vaccine caused a dramatic reduction in the incidence of invasive pneumococcal diseases in American children, reduced antibiotic use and the number of medical visits due to otitis media and pneumonia by children. The incidence of invasive pneumococcal diseases caused by resistant strains declined in immunized children, adults and elderly individuals. In Brazil, the mortality rate of pneumococcal meningitis is very high and the resistance to antibiotics has increased over the last 5 years. CONCLUSIONS: Heptavalent pneumococcal conjugate vaccine can benefit not only children, but the entire community and should be included in the Brazilian routine immunization schedule. PMID- 16721441 TI - Vaccines under development: group B streptococcus, herpes-zoster, HIV, malaria and dengue. AB - OBJECTIVES: To review the current state of development of streptococcus B, herpes zoster, HIV, malaria and dengue vaccines. These vaccines were selected both because of imminent commercial release and because of specific problems with their development. SOURCES OF DATA: A review of the literature was performed by means of a MEDLINE search, on the period 1996 to 2006, for the epidemiology and immunology of these diseases, analyzing both the greatest obstacles to creating a vaccine and the current state of research, with emphasis on studies in the most advanced stages. SUMMARY OF THE FINDINGS: Each of the five diseases chosen presents specific problems for vaccine development. Nevertheless, in the majority of cases these have been or are in sight of being resolved, allowing for the prediction that a safe and effective vaccine - or vaccines - will be available in the near future. CONCLUSIONS: Despite the problems faced in developing these vaccines, advances in molecular biology and immunology have made it possible to overcome most obstacles, opening up the prospects for new vaccines. PMID- 16721442 TI - [Abraham Horwitz Barak]. PMID- 16721443 TI - [Clinical and epidemiological characteristics of respiratory virus infections among adults hospitalized during 2004 influenza season]. AB - BACKGROUND: During yearly influenza (FLU) season, FLU viruses are well represented among hospitalized patients as in the community. Also, other respiratory viruses could be represented among adult in-patients. AIM: to describe the presence and clinical- epidemiological characteristics of non-FLU respiratory virus infections (respiratory syncytial-RSV, parainfluenza and adenovirus-ADV) among hospitalized adults during FLU season and to compare with FLU-A (IA) or -B (IB) cases. PATIENTS AND METHODS: Adult patients hospitalized at Hospital Clinico Universidad Catolica between May to July 2004 with a respiratory virus infections confirmed by rapid antigen test or direct immunofluorescence of IA, IB (Flu group) or RSV, parainfluenza (1-2-3) and ADV (non-Flu group) were included. RESULTS: 86 cases were identified: 73.5% FLU (48.2% IA, 25.3% IB) and 26.5% non-FLU (15.7% parainfluenza-2; 8.4% RSV; 1.2% parainfluenza-3; 1.2% ADV). No differences were observed in general characteristics and evolution of patients. In FLU-group were more frequently observed myalgia, cough, hospitalization due to febrile syndrome, higher values of C-reactive protein and band leukocytes count (p < 0.05). CONCLUSIONS: During 2004 FLU season a 26.5 % of respiratory viral infection were due to non-FLU viruses among adult hospitalized in our centre. The difficulty to difference FLU vs. non-FLU infections, suggest that it is necessary to include other respiratory virus in the viral etiological diagnosis, even in FLU season. PMID- 16721444 TI - [Respiratory virus infections in children with acute lymphoblastic leukemia and febrile neutropenia: a prospective study]. AB - Respiratory viruses are the most common infections in healthy children. The impact of these infections in cancer patients has been only recently recognized in Chile. AIM: To establish the frequency and epidemiological-clinical profile of respiratory virus infections in children younger than 15 years of age with acute lymphoblastic leukemia requiring hospitalization due to a febrile neutropenic episode. METHODS: All children < 15 years of age requiring hospitalization in the Regional Hospital of Valdivia, Chile, with cancer, fever and neutropenia between November 1 2002 and October 31 2004 were studied. Nasopharyngeal aspirate were obtained and tested by direct immunofluorescent assays for influenza A-B virus, parainfluenza virus type 1, 2, 3, respiratory syncytial virus (RSV) and adenovirus. RESULTS: Respiratory viruses were detected in 25% of 44 febrile neutropenia episodes occurring in 25 patients. Viruses detected were, influenza (3/11), parainfluenza (3/11) RSV (2/11), adenovirus (1/11), parainfluenza + RSV (1/11), and parainfluenza + adenovirus (1/11). Four (36%) of cases were nosocomial. Respiratory symptoms were present in 9 children, seven of whom had inferior respiratory tract symptoms. One patient died of persistent RSV infection, agranulocytosis and extensive bilateral pneumonia. CONCLUSIONS: Respiratory viruses were detected in 25% of febrile neutropenic episodes in children with ALL. This high detection rate together with the frequent involvement of the lower respiratory tract and the possibility of death strongly favors the inclusion of respiratory virus diagnostic tests in the evaluation of these children during autumn and winter season. PMID- 16721445 TI - [Rotavirus Vaccine. Statement of the Consultive Committee of Immunizations on behalf of The Chilean Infectious Diseases Society. March 2006]. AB - The article briefly reviews the epidemiology of rotavirus infection and the scientific information of the rotavirus vaccines: Rotashield, withdrawn from the market due to its association with intussusception, Rotateq currently in an advanced phase of development, and Rotarix, recently licensed in Chile. Considering the available information, the Consultive Committee of Immunizations of the Chilean Society of Infectious Diseases, summarizes its conclusions and makes recommendations for infants vaccination against rotavirus in our country. PMID- 16721446 TI - [Acute respiratory infections in children of day care center: characteristics and costs]. AB - The clinical profile of respiratory infections in 140 children who attend a day care center was described in a prospective study and direct costs were estimated. The costs estimation included diagnostic procedures, laboratory exams, drugs, kinesitherapy, parents' absenteeism, medical certificates and hospitalization. Two hundred and two respiratory episodes in 90 (64.2%) children, 75 (37.1%) in 30 infants and 127 (62.8%) in 60 toddlers were analyzed. There were 61.3% lower respiratory infections in infants versus 39.4% in toddlers (p < 0.002). Antimicrobials were prescribed in 42.6% of the medical attendance. Infants represent major costs regarding medical consultations, kinesitherapy, laboratory tests and refunds for medical certificates or leave of absence (p < 0.001). The mean global cost of a respiratory infection was US $ 129,3 for infants and US $ 53,0 for toddlers (p < 0.001). This situation needs the development of an intervention program. PMID- 16721447 TI - [Tuberculous meningitis in adults: review of 53 cases]. AB - Tuberculous Meningitis (TBM) is the most severe form of extrapulmonary tuberculosis. The clinical spectrum is broad and may be non-specific making early diagnosis difficult. This increases the incidence of mortality. We describe the clinical characteristics of patients with TBM in Dr. Lucio Cordova's Infectious Disease Hospital in Santiago, Chile, between 1995 and 2002. We review 53 adult cases of TBM, with a median age of 39 years. At admission 66% of the patients had some mental status deterioration, and the classic triad of symptoms of meningeal irritation was present only in 30%. The cerebrospinal fluid (CSF) examination showed increased protein level, low glucose level and lymphocytic pleocytosis in most. Thirty percent of the patients were coinfected with HIV. The mortality in the later group was greater than in the TBM population as a whole (31 vs 17%). TBM is still a present diagnostic problem, in spite of the new diagnostic methods. A high index of suspicion is required in order to make an early diagnosis. PMID- 16721448 TI - [Brain abscess: about a clinical series of 30 patients]. AB - A retrospective study was designed to ascertain the main clinical features of 30 patients admitted for brain abscess between 1989 and 2005. A contiguous source of infection was identified in 40% of cases, direct inoculation secondary to trauma or neurosurgery in 23,3%, and a distant source in 23,3%. Fever, headache and sensorial involvement were the main clinical manifestations. Temporal and frontal lesions were predominant. Microbiological information was available in only half of cases, prevailing polymicrobial and gram-positive bacterial infections. Most patients underwent surgical procedures (66.7%) and the rest received only medical therapy. During antimicrobial treatment, 6 patients (20%) suffered a serious adverse event that prompted discontinuation or change of therapy. Failure of treatment was observed in 40%, including 3 deaths (10%), non-programmed surgical drainage or reintervention (16.7%) and changes of the antimicrobial regimen (13.3%). Eight patients were discharged with sequels (26.7%), and during follow up, 21% presented major events (death or brain abscess-related readmission). PMID- 16721449 TI - [Splenic abscess: presentation of nine cases]. AB - Relatively rare, splenic abscess is difficult to diagnose and often fatal if left untreated. The disease is thought to be growing in frequency because of the increasing number of inmunocompromised patients. Several mechanisms for the development of splenic abscess may exist. Some studies demonstrate that prior splenic injury in addition to bacteraemia is required for a splenic abscess to occur. In our series, 9 non immunocompromised patients were identified to have this disease during a 6 years period. Pathogens isolated included Salmonella sp, Staphylococcus sp and Enterococcus sp. Splenectomy was performed in three patients; in another a percutaneous drainage was done. One patient died. In summary, though rare, splenic abscess presents with high morbidity and mortality. In our experience, risk factors as immunocompromise seem not to be so prevalent in patients with splenic abscess and therefore this diagnosis should be considered in all patients with fever of unknown origin. PMID- 16721450 TI - [Rhodococcus equi]. PMID- 16721451 TI - Vaccines against papillomavirus infections and disease. PMID- 16721452 TI - [Flu vaccine in children: state of the art]. AB - Influenza is an acute respiratory illness that year to year causes considerable morbidity and mortality. . All children, healthy or at high risk, are susceptible to this infection. Influenza vaccine seems to be effective in preventing influenza-like illness, laboratory-confirmed infection, and hospitalizations and deaths, demonstrating cost-effectiveness. Inactivated and live-attenuated vaccines have a similar efficacy profile. . Due to high influenza associated hospitalization rates in children, some countries have recommended incorporating influenza vaccination into the universal immunization program for healthy infants between 6-24 months of age. Considering that this public-health policy has been incorporated in Chile, local studies assessing its impact are needed. PMID- 16721453 TI - [The history of penicillin and of its manufacturing in Chile]. PMID- 16721455 TI - Shock of paradigms on the instrumentation of curved root canals. AB - This paper makes a practical analysis about the paradigm on the instrumentation of curved root canals. In Endodontics, a paradigm has been created. Theories and techniques for instrumentation of curved root canals state that the use of a #25 file in the apical portion fulfills all the requirements for cleaning and shaping of the root canal system. Every scientific theory or paradigm should be continuously opened to modifications or refutation. The existence of extremely flexible instruments fabricated from metal alloys, methods for accurate determination of the real anatomic diameter and achievement of optimal shaping and cleaning of the apical portion created new theories and a new paradigm on the instrumentation of curved root canals. Therefore, this new insight will gradually modify the mentality of researchers and clinicians, but will still be open to further investigations and theories. PMID- 16721456 TI - The importance of apical patency and cleaning of the apical foramen on root canal preparation. AB - The apical limit of root canal instrumentation has always been a matter of great controversy. Despite the large number of published studies on this subject, a consensus has not yet been reached. In fact, the recent discussion on apical patency and cleaning of the apical foramen, as well as the incorporation of these procedures to the endodontic treatment, seem to have raised even more polemics. It is likely that all this polemics has its roots in the lack of interrelation between the theoretical knowledge of pulp stump and periapical tissues and the real clinical practice. By addressing the most important aspects of this theme, this paper aims to present news concepts about the importance of apical patency and cleaning of the apical foramen during root canal preparation. PMID- 16721457 TI - NSK reciprocating handpiece: in vitro comparative analysis of dentinal removal during root canal preparation by different operators. AB - The purpose of this study was to assess dentin removal during root canal preparation by different operators using a NSK reciprocating handpiece. Eighty four human single-rooted mandibular premolars were hand instrumented using Triple Flex stainless-steel files (Kerr) up to #30, weighed in analytical balance and randomly assigned to 4 groups (n=21). All specimens were mechanically prepared at the working length with #35 to #45 Triple-Flex files (Kerr) coupled to a NSK (TEP E10R, Nakanishi Inc.) reciprocating handpiece powered by an electric motor (Endo Plus; VK Driller). Groups 1 to 4 were prepared by a professor of Endodontics, an endodontist, a third-year dental student and a general dentist, respectively. Teeth were reweighed after root canal preparation. The difference between weights was calculated and the means of dentin removal in each group were analyzed statistically by ANOVA and Tukey's test at 5 % significance level. The greatest amount of dentin removal was found in group 4, followed by groups 2, 3 and 1. Group 4 differed statistically from the other groups regarding dentin removal means [p<0.001 (group 1); p=0.005 (group 2); and p=0.001 (group 3)]. No statistically significant difference was found between groups 1 and 2 (p=0.608), 1 and 3 (p=0.914) and 2 and 3 (p=0.938). In conclusion, although the group prepared by a general dentist differed statistically from the other groups in terms of amount of dentin removal, this difference was clinically irrelevant. The NSK reciprocating handpiece powered by an electric engine was proved an effective auxiliary tool in root canal preparation, regardless of the operator's skills. PMID- 16721458 TI - Molecular analysis of Er:YAG laser irradiation on dentin. AB - The aim of this study was to evaluate by dispersive Raman spectroscopy the mineral and organic components of human dentin before and after laser irradiation and acid etching. The occlusal enamel of six non-carious human third molars was removed providing 6 dentin discs, which were divided in four quadrants each of them receiving a different surface treatment: etching with a 37% phosphoric acid gel (control); irradiation by Er:YAG laser (KaVo Key Laser II) with 80 mJ, 3 Hz, 30 s (group I); 120 mJ, 3 Hz, 30 s (group II); and 180 mJ, 3 Hz, 30 s (group III). The Raman spectra of normal (untreated) and treated dentin were analyzed and the mineral and the organic component were evaluated. Results were submitted to statistical analysis by ANOVA and Tukey's test at 5% significance level. The minerals and organic content were less affected in the control group and group I (p>0.05). Group II presented more reduction in mineral content (p<0.01) whereas in group III the inorganic (p<0.05) and organic (p<0.01) content were more affected. Dispersive Raman spectroscopy provided valid information of dentin chemical constituents with non-chemical sampling preparation. PMID- 16721459 TI - Flexural strength of pure Ti, Ni-Cr and Co-Cr alloys submitted to Nd:YAG laser or TIG welding. AB - Welding of metals and alloys is important to Dentistry for fabrication of dental prostheses. Several methods of soldering metals and alloys are currently used. The purpose of this study was to assess, using the flexural strength testing, the efficacy of two processes Nd:YAG laser and TIG (tungsten inert gas) for welding of pure Ti, Co-Cr and Ni-Cr alloys. Sixty cylindrical specimens were prepared (20 of each material), bisected and welded using different techniques. Four groups were formed (n=15). I: Nd:YAG laser welding; II- Nd:YAG laser welding using a filling material; III- TIG welding and IV (control): no welding (intact specimens). The specimens were tested in flexural strength and the results were analyzed statistically by one-way ANOVA. There was significant differences (p<0.001) among the non-welded materials, the Co-Cr alloy being the most resistant to deflection. Comparing the welding processes, significant differences (p<0.001) where found between TIG and laser welding and also between laser alone and laser plus filling material. In conclusion, TIG welding yielded higher flexural strength means than Nd:YAG laser welding for the tested Ti, Co-Cr and Ni Cr alloys. PMID- 16721460 TI - Bond strength of three dental porcelains to Ni-Cr and Co-Cr-Ti alloys. AB - Ceramometal bond strength has played an important role for the replacement of gold alloys by nickel-chromium alloys in dentistry. This study evaluated the metal/porcelain bond strength of three ceramic systems (Vita VMK 88, Williams and Duceram) associated with three nickel-chromium alloys (Durabond, Lite Cast B and Resistal P) and one experimental cobalt-chromium-titanium alloy. Thirty cast cylinder specimens (15 mm in height; 6 mm in diameter) were obtained for each alloy, in away that 10 specimens of each alloy were tested with each porcelain. Bond strength was measured with an Emic screw-driven mechanical testing machine by applying parallel shear forces to the specimens until fracture. Kruskal-Wallis and Mann-Whitney U tests were used for statistical analysis of the alloy/ceramic combinations (p<0.05). Resistal P/Duceram had significantly higher bond strength (44.38+/-9.12 MPa) (p<0.05) than the other combinations, except for Co-Cr-Ti alloy/Vita VMK 88 (38.41+/-12.64 MPa). The association of the experimental Co-Cr Ti alloy with Williams porcelain had significantly higher bond strength (28.20+/ 3.86 MPa) than the combination of other alloys with the same porcelain (p<0.05). Based of these results and within the limitations of an in vitro study, it may be concluded that the bond strength of the three ceramic systems to the Ni-Cr and Co Cr-Ti alloys varied significantly, indicating that metal/ceramic compatibility was very important to the bond strength. PMID- 16721461 TI - Abrasion resistance of composites polymerized by light-emitting diodes (LED) and halogen light-curing units. AB - This study compared the abrasion resistance of direct composite resins cured by light-emitting diodes (LED) and halogen light-curing units. Twenty specimens (12 mm in diameter; 1.0 mm thick) of each composite resin [TPH (Dentsply); Definite (Degussa); Charisma (Heraus Kulzer)] were prepared using a polytetrafluoroethylene matrix. Ten specimens per material were cured with the LED source and 10 with the halogen lamp for 40 s. The resin discs were polished, submitted to initial surface roughness reading (Ra initial - microm) in a roughness tester and stored in water at 37 degrees C for 15 days. The specimens were weighed (M1) and submitted to simulated toothbrushing using slurry of water and dentifrice with high abrasiveness. After 100 minutes in the toothbrushing simulator, the specimens were cleaned, submitted to a new surface roughness reading (Ra final - microm) and reweighed (M2). Mass loss was determined as the difference between M1 and M2. Data were recorded and analyzed statistically by one-way ANOVA and Tukey Test at 5% significance level. The composite resin with greater size of inorganic fillers (TPH) showed the lowest mass loss and surface roughness means, indicating a higher resistance to toothbrush abrasion (p<0.05). Definite cured with LED presented the least resistance to toothbrush abrasion, showing the highest means of surface roughness and mass loss (p<0.05). The LED source did not show the same effectiveness as the halogen lamp for polymerizing this specific composite resin. When the composite resins were cured a halogen LCU, no statistically significant difference was observed among the materials (p>0.05). It may be concluded that the type of light-curing unit and the resin composition seemed to interfere with the materials' resistance to abrasion. PMID- 16721462 TI - Residual monomer in a composite resin after light-curing with different sources, light intensities and spectra of radiation. AB - The purpose of this study was to investigate the amount of residual monomer in a composite resin after light-curing with different sources, light intensities and spectra of radiation. The resin specimens (4 mm in diameter; 2 mm thick) (n=5) were inserted in Plexglass matrixes and light-cured with a halogen lamp, LED and PAC units for 40, 40 and 5 s, respectively. The polymerized specimens were ground and 25 mg of each specimen were immersed in 8 mL 96% ethanol for 24 h to extract the residual monomer. The amount of residual monomer was determined by the standard straight line method. The light spectrum of the different LCUs was shot and processed through researches of the absorbation of the standard solutions at a level of lambda=275 nm and their extracts. Data were analyzed statistically by variational dispersion analysis and Tukey-Kramer test at 5% significance level. It was observed that, the halogen lamp produced the smallest amount of monomer under sufficient light intensity. The spectrum of light radiation of PAC was within the limits of 450- 490 nm and was of extremely high intensity. This imposes the use of short-limited light impulses, but also might lead to deterioration quality of polymerization. The LED unit had the best spectral radiation. An increase of light intensity was proved necessary. PMID- 16721463 TI - Shear bond strength of acetone-based one-bottle adhesive systems. AB - The aim of this study was to assess the shear bond strength of four acetone-based one-bottle adhesive systems to enamel and dentin, and compare to that of an ethanol-based system used as control. Fifty human molars were bisected mesiodistally and the buccal and lingual surfaces were embedded in acrylic resin using PVC cylinders. The buccal surfaces were ground to obtain flat dentin surfaces, while the lingual surfaces were ground to obtain flat enamel surfaces. All specimens were polished up to 600-grit sandpapers and randomly assigned to 5 groups (n=20; 10 dentin specimens and 10 enamel specimens), according to the adhesive system used: One-Step (Bisco); Gluma One Bond (Heraeus Kulzer); Solobond M (Voco); TenureQuik w/F (Den-Mat) and OptiBond Solo Plus (Kerr) (control). Each adhesive system was applied according to the manufacturers' instructions. The respective proprietary hybrid composite was applied in a gelatin capsule (d=4.3 mm) and light-cured for 40 s. The specimens were tested in shear strength with an Instron machine at a crosshead speed of 5 mm/min. Bond strengths means were analyzed statistically by one-way ANOVA and Duncan's post-hoc (p< or =0.05). Shear bond strength means (MPa) (+/-SD) to enamel and dentin were: Enamel: One Step=11.3(+/-4.9); Gluma One Bond=16.3(+/-10.1); Solobond M=18.9(+/-4.5); TenureQuik w/F=18.7(+/-4.5) and OptiBond Solo Plus=16.4(+/-3.9); Dentin: One Step=6.4(+/-2.8); Gluma One Bond=3.0(+/-3.4); Solobond M=10.6(+/-4.9); TenureQuik w/F=7.8(+/-3.9) and OptiBond Solo Plus=15.1(+/-8.9). In enamel, the adhesive systems had statistically similar bond strengths to each other (p>0.05). However, the ethanol-based system (OptiBond Solo Plus) showed significantly higher bond strength to dentin than the acetone-based systems (p< or =0.0001). In conclusion, the solvent type (acetone or ethanol) had no influence on enamel bond strength, but had great influence on dentin bonding, which should be taken into account when choosing the adhesive system. PMID- 16721464 TI - Shear bond strength of new and recycled brackets to enamel. AB - The purpose of this study was to evaluate in vitro the shear bond strength of recycled orthodontic brackets. S2C-03Z brackets (Dental Morelli, Brazil) were bonded to the buccal surfaces of 50 extracted human premolars using Concise Orthodontic chemically cured composite resin (3M, USA). The teeth were randomly assigned to 5 groups (n=10), as follows. In group I (control), the bonded brackets remained attached until shear testing (i.e., no debonding/rebonding). In groups II, III and IV, the bonded brackets were detached and rebonded after recycling by 90-microm particle aluminum oxide blasting, silicon carbide stone grinding or an industrial process at a specialized contractor company (Abzil Lancer, Brazil), respectively. In group V, the bonded brackets were removed and new brackets were bonded to the enamel surface. Shear bond strength was tested in an Instron machine at a crosshead speed of 0.5 mm/min. Data were analyzed statistically by ANOVA and Tukey's test at 5% significance level. There was no statistically significant difference (p>0.05) between the control brackets (0.52 kgf/mm2), brackets recycled by aluminum oxide blasting (0.34 kgf/mm2) and new brackets attached to previously bonded teeth (0.43 kgf/mm2). Brackets recycled by the specialized company (0.28 kgf/mm2) and those recycled by silicon carbide stone grinding (0.14 kgf/mm2) showed the lowest shear strength means and differed statistically from control brackets (0.52 kgf/mm2) (p<0.05). In conclusion, the outcomes of this study showed that bracket recycling using 90-microm aluminum oxide particle air-abrasion was efficient and technically simple, and might provide cost reduction for orthodontists and patients alike. PMID- 16721465 TI - In vitro evaluation of microleakage of different materials used as pit-and fissure sealants. AB - The aim of this study was to evaluate in vitro the marginal microleakage of different materials used as pit-and-fissure sealants (Delton, Filtek Flow, Dyract Flow and Vitremer). Fifty-six extracted sound human third molars were randomly assigned to 4 groups (n=14). After sealant placement, the teeth were thermocycled (500 cycles; 5 degrees C, 37 degrees C and 55 degrees C), isolated, immersed in 2% buffered methylene blue dye for 4 h, included in acrylic resin and sectioned longitudinally in a buccolingual direction. The sections were analyzed for leakage using an stereomicroscope. A 4-criteria ranked scale was used to score dye penetration. All materials exhibited dye penetration to some extension and no statistically significant difference was observed among the groups (p>0.05). In conclusion, the findings of this study showed that a flowable composite resin, a flowable compomer and resin-modified glass ionomer placed on occlusal pits and fissures had similar marginal sealing as the unfilled self-cured resin-based sealant. PMID- 16721466 TI - Effect of water storage on tooth displacement in maxillary complete dentures. AB - The purpose of this study was to investigate the effect of water storage at 37 degrees C (1 week, 1 month and 3 months) on tooth displacement in maxillary complete dentures. Ten maxillary dentures were constructed with Classico acrylic resin using the conventional method of packing in metallic flasks. Metallic reference pins were placed in the incisal border of the central incisors (I), labial cusp of the first premolars (PM), and mesiolabial cusp of the second molars (M). Twelve hours after final flask closure, the acrylic resin was cured in water at 74 degrees C for 9 h. The flasks were removed from the thermo polymerizing unit after water-cooling and the dentures were deflasked, finished and stored in water at a temperature of 37 degrees C for 1 week, 1 month and 3 months. At deflasking and at the water storage intervals, the I-I (incisor to incisor), PM-PM (pre-molar to pre-molar), and M-M (molar to molar) transversal distances, and LI-LM (left incisor to left molar) and RI-RM (right incisor to right molar) anteroposterior distances were measured using an optical microscope with 0.0005 mm accuracy. Data were submitted to ANOVA and Tukey's test (5%). Comparing the evaluation periods for each individual transversal and anteroposterior reference point, no statistically significant differences were observed among deflasking and the water storage intervals for I-I, PM-PM, M-M and RI-RM distances (p>0.05). For LI-LM, however, deflasking values were statistically different from those of 1-week, 1-month and 3-month water storage intervals (p<0.05), which, in turn, did not differ statistically to each other (p>0.05). These results confirm the complexity of tooth displacement in complete dentures. From a clinical standpoint, the difference observed in LI-LM distance after water storage would not be detected by the patients during clinical use. PMID- 16721467 TI - Toxicity screening of mouthwashes in the pollen tube growth test: safety assessment of recommended dilutions. AB - This study investigated whether mouthwash dilutions, recommended by the manufacturers, may contribute to oral mucosa irritation. Twenty frequently used mouthwash brands were examined by the pollen tube growth test, an alternative in vitro method for assessment of the irritating potential of ingredients of cosmetic formulations. The test is based on the photometric quantification of pollen tube growth inhibition. This parameter is expressed by IC50 values that characterize the cytotoxic potential of a product. The IC50s clearly revealed that none of the tested mouthwashes should cause acute irritation if used in the recommended dilution. However, at least 4 of the mouthwashes evaluated in this study could probably irritate the oral mucosa acutely if used undiluted (original concentration). PMID- 16721468 TI - Effectiveness and efficiency of chemomechanical carious dentin removal. AB - The aims of this in vitro study were both to determine the time necessary for removal of carious dentin (efficiency) and the Knoop Hardness Number (KHN) of the remaining dentin (effectiveness), using a chemomechanical method (Carisolv) or hand excavation. Thirty human molars were bisected through occlusal carious lesions into two equal halves. Each half was randomly excavated by hand in circular movements with a spoon excavator or using Carisolv gel according to the manufacturer's instructions. The duration of carious dentin removal was recorded. Tooth sections were resin-embedded, ground flat and polished. Dentin KHN was determined at distances of 100, 200, 300, 400 and 500 microm from the cavity floor. Data were analyzed by Wilcoxon's test (alpha= 0.01), ANOVA and Student's t test (alpha= 0.05). The median of the time necessary for chemomechanical excavation was significantly greater than for hand excavation. KHN means (+/- SD) at 100, 200, 300, 400, 500 microm for chemomechanical method were, respectively: 15.6 (+/- 4.96), 18.0 (+/- 6.22), 21.3 (+/- 9.30), 24.3 (+/- 9.25), 28.5 (+/- 11.80); and for hand excavation were: 21.2 (+/- 10.26), 23.4 (+/- 9.49), 28.2 (+/ 11.62), 31.0 (+/- 12.17), 34.3 (+/- 11.95). It may be concluded that hand excavation presented higher efficiency and effectiveness than chemomechanical excavation. PMID- 16721469 TI - Anterior open bite--cephalometric evaluation of the dental pattern. AB - The purpose of this study was to compare the dental pattern of patients with anterior open bite malocclusion to that of individuals with normal overbite by utilization of lateral cephalograms, panoramic radiographs and study casts. The findings showed that there was no significant difference in the inclination of the occlusal plane (SN.PlO) and position of the maxillary and mandibular incisors (1-NA, 1-NB) between both groups of individuals; but the angles of inclination of the maxillary and mandibular incisors (1.1, 1.NA and 1.NB) differed statistically between patients with anterior open bite of the individuals that presented normal overbite, which suggests that the anterior open bite may be of dental origin. PMID- 16721470 TI - Anatomosurgical study of the marginal mandibular branch of the facial nerve for submandibular surgical approach. AB - The purpose of this study was to estimate the distance from the mandibular marginal branch of the facial nerve to the inferior margin of the mandible in order to determine the best and safest location to approach the posterior mandibular region. Forty-five hemi-faces of 27 Brazilian adult cadavers were dissected and the distance between the mandibular marginal branch and the inferior margin of the mandible was measured. The number of marginal branches and anastomoses with other branches of the facial nerve was also recorded. The evaluation of the anatomic pieces showed 1 to 3 branches of the marginal mandibular branch, anastomoses with the buccal and cervical branches of the facial nerve and distances between 1.3 cm to +1.2 cm from the inferior margin of the mandible. In 57.7% of the cases, the nerve passed superiorly and along the length of the inferior margin of the mandible. Based on the findings of the present anatomosurgical study, it may be recommended an incision 3 cm below the inferior margin of the mandible associated to a careful dissection in planes and flap retraction. This is expected to reduce the risk of neuropraxia of the marginal mandibular nerve making the submandibular incision a safe approach. PMID- 16721471 TI - Root canal therapy of a maxillary first molar with five root canals: case report. AB - This paper reports the case of a maxillary left first molar that presented three root canals in the mesiobuccal root. Root canal therapy and case management are described. Features like wide crown access, adequate illumination and use of exploring files where important for successful completion of the endodontic treatment. PMID- 16721472 TI - Oral focal epithelial hyperplasia: report of five cases. AB - Focal epithelial hyperplasia or Heck's disease is a rare contagious disease caused by human papillomavirus types 13 or 32, initially described among Native American populations. This condition is characterized by the occurrence of multiple small papules or nodules in oral cavity, especially on labial and buccal mucosa and tongue. This report describes the diagnosis of focal epithelial hyperplasia in five Central Amazonian Indians who sought treatment at the Amazonas State Foundation of Tropical Medicine (FMT-AM), using clinical criteria, polymerase chain reaction (PCR) and DNA sequencing. PMID- 16721473 TI - Complicated crown fracture: a case report. AB - This case report describes the treatment of a severe crown fracture of an immature mandibular permanent incisor in a 12 year-old boy referred to the Dental Traumatology Center of UNIGRANRIO University. After conventional endodontic procedures and dressing with calcium hydroxide, an apical plug was made with mineral trioxide aggregate in order to avoid an overflow of gutta-percha and endodontic sealer into the periapical tissues. Two years later, the tooth was clinically asymptomatic and radiographs showed total repair of the apical lesion. PMID- 16721477 TI - [Magnetic resonance in moderate and severe head injury: comparative study of CT and MR findings. Characteristics related to the presence and location of diffuse axonal injury in MR]. AB - INTRODUCTION: Cranial CT has been the most extended evaluation means for patients suffering head trauma. However, it has low sensitivity in the identification of diffuse axonal injury and posterior fossa lesions. Cranial MR is a potentially more sensitive test but difficult to perform in these patients, a fact that has hampered its generalised use. OBJECTIVE: To compare the identification capability of traumatic intracranial lesions by both diagnostic tests in patients with moderate and severe head injury and to determine which radiological characteristics are associated with the presence of diffuse injury in MR and their clinical severity. MATERIAL AND METHODS: 100 patients suffering moderate or severe head injury to whom a MR had been performed in the first 30 days after trauma were included. All clinical variables related to prognosis were registered, as well as the data from the initial CT following Marshall et al., classification. The MR was blindly evaluated by two neuroradiologists that were not aware of the initial CT results or the clinical situation of the patient. All lesions were registered as well as the classification following the classification of lesions related to DAI described by Adams et al. CT and MR findings were compared evaluating the sensitivities of each test. Factors related to the presence of diffuse injury in MR were studied by univariate analysis using chi2 test and simple correlations. RESULTS: MR is more sensitive than CT for lesions in cerebral white matter, corpus callosum and brainstem. It also detects a greater number of cerebral contussions. The presence of diffuse axonal injury depends on the mechanism of the trauma, being more frequent in higher energy trauma, specially in traffic accidents. Among the radiological characteristics associated to DAI the most clearly related is intraventricular haemorrhage. The presence of a deeper injury and a higher score in the scales of Adams is associated with a lower score in the GCS and motor GCS, and so with a worse level of consciousness and bigger severity of injury, confirming Ommaya's model. PMID- 16721478 TI - [Postoperative delirium in patient neurosurgical: evaluation by means of the Abbreviated Mental Test]. AB - OBJECTIVES: To assess the incidence and characteristics of postoperative changes in the higher cerebral functions after elective intracranial surgery under general anesthesia. PATIENTS AND METHODS: This is a prospective study of 60 patients, aged 18-81 years, submitted to neurosurgical operations, allocated into two groups of 30 patients each: intracranial surgery group, patients submitted to craniotomy, and extracranial surgery group or control group, patients submitted to spinal procedures. All patients were given the Abbreviated Mental Test (AMT) on the day before to the operation, and then 2 and 24 hours after the end of the procedure. All individuals were managed with the same anesthetic technique. RESULTS: No differences in regard to demographics, duration of the operation, ASA physical state, and habits were found between the two groups. No differences in the pre-surgical and post-surgical AMT mean score were encountered between patients submitted to intracranial 9.87 +/- 0.35 or to spinal surgeries 9.80 +/- 0.41. Similarly, there were no significant differences between the two groups in the results of the AMT performed at 2 and 24 hours after the end of the surgeries. Only two subjects undergoing spinal procedures had a score of < or = 8 in the AMT performed 2-hours after the operation, while none showed a decrease in the 24-hour test score. CONCLUSIONS: Patients submitted to intracranial surgery did not show any changes in cognitive or attention functions during the first postoperative 24 hours as assessed by the AMT. PMID- 16721479 TI - [Spinal meningiomas]. AB - OBJECTIVE: To present the incidence, clinical presentation, radiological features, localization, surgical techniques, and long term results of surgically treated spinal meningiomas. PATIENTS AND METHODS: We review retrospectively 57 spinal meningiomas in 55 patients operated between January 1980 and June 2004. Age, sex, duration and type of symptoms, radiological and surgical results, histological features and long term outcome are analyzed. RESULTS: Fifty five patients, consisting 46 females and 9 males, with an age range from 12 to 84 years (mean 65.2+/-9.2) were identified. The first symptom was motor deficit (N=34, 61.8%) and the mean duration symptoms were: 6.2 months. Distribution along the spinal axis was 51 thoracic, 5 cervical and 1 lumbar. The most common type was meningothelial (N=31, 54.3%). CONCLUSIONS: Meningiomas account in our experience, the 46% of spinal cord tumours and account the 7.5% of all meningiomas. They are more frequent in females most frequently occur in the thoracic region and have low recurrence rate. Typically, they are located in the intradural extramedullary space, grow slowly, and spread laterally in the subarachnoid space. Finally we concluded that magnetic resonance imaging is the best imaging technique for diagnosis and total tumour resection improved the surgical results of spinal meningiomas. PMID- 16721480 TI - [Anterior cervical fusion with tantalum interbody implants. Clinical and radiological results in a prospective study]. AB - Anterior cervical discectomy and interbody fusion (ACDF) is a widely accepted surgical technique in the treatment of cervical disc disease. Tantalum cages have been recently introduced in spine surgery for interbody fusion because of the advantages of their mechanical properties. We present the results of a prospective clinical and radiological study on 24 consecutive patients who underwent an ACDF with tantalum cages. Clinical evaluation was assessed preoperatively and after surgery by a questionnaire that included a Visual Analogic Scale (VAS) of neck and arm pain, the Oswestry Disability Index and the Zung Depression Scale. Results were classified by Odom's criteria. Radiological evaluation included flexion-extension X-rays, and changes in distance between spinous processes and Cobb angle were measured. Postoperatively patients were reviewed 3 and 12 months after surgery. A statistical significative improvement in all clinical data was reported. According to Odom's criteria in 75% of patients the results were considered like excellent or good. Only one case of radiological and clinical pseudoarthrosis was confirmed. No significative differences were reported 3 and 12 months after surgery. Tantalum cages are a very promising and usefull alternative among implants available for ACDF. Compatibility with MRI postoperative studies and the unnecessariness of autograft are some of their advantages. PMID- 16721481 TI - [Development of the anterior cervical postdiscectomy arthrodesis: bone graft, plate, intersomatic cage and plate-cage]. AB - The surgical techniques for anterior cervical postdiscectomy arthrodesis are reviewed, including bone graft, cervical plates, intervertebral cages and plate cage systems. The biomechanical advantages of each method are discussed. An alternative to arthrodesis are simple discectomy and arthroplastia with disc prosthesis. Integrated plate-cage systems are the final result in the evolution of the cervical arthrodesis developments. The available plate-cage systems are described. PMID- 16721482 TI - [Aesthetic and visual dynamics of the presentation. Basic concepts]. AB - Aesthetic and dynamic aspects of scientific presentations have been scarcely discussed in the medical literature. In this work, basic concepts regarding colour, management, hierarchical disposition of the text, and graphics' animations are briefly described to allow an harmonic and balanced exposition of the visual presentation. PMID- 16721483 TI - [Cranial hemophilic pseudotumor]. AB - The haemophilic pseudotumor is defined as an encased hematoma that increases of volume progressively by episodes of recurrent hemorrhage. It is a rare complication of haemophilia occurring in 1-2% of patients with moderate or severe factor VIII or IX deficiency. Its more frequent location is in the long bones of low extremities and pelvis. We report a case of a 21-year-old man with moderate deficiency of factor VIII (19% of normal factor VIII activity) that developed a pseudotumor in the cranium. To our knowledge, this is the third case of the cranial hemophilic pseudotumor in medical literature. PMID- 16721484 TI - First illustrations of female "Neurosurgeons" in the fifteenth century by Serefeddin Sabuncuoglu. AB - Males have dominated medicine for many centuries. Females could not appear in the medical history equally till the end of the 18th century; although they always have been in medicine as healers. It is worth mentioning that first illustrations indicating female surgeons were found in the book written in Turkish by Serefeddin Sabuncuoglu in the 15th century; while Europe was newly waking up from its dark ages and Middle East was under the influence of strict rules of Arabic and Islamic culture. Serefeddin Sabuncuoglu (1385-1470) was the author of the first illustrated surgical textbook Cerrahiyyetu'l-Haniyye (Imperial surgery) in Turkish Literature. Inside miniatures drawn by Serefeddin Sabuncuoglu indicated that the female surgeons, acknowledged "Tabibe", had been allowed to practice alone in Anatolia. Tabibes are illustrated in the miniatures practicing on the management of dead foetus with foetal hydrocephalus and macrocephalus which were the first clues by means of Turkish women in Neurosurgery. PMID- 16721485 TI - A novel 111/121 diplotype in the Calpain-10 gene is associated with type 2 diabetes. AB - Genetic variations in the Calpain-10 gene, CAPN10, have been reported to be associated with the risk of type 2 diabetes mellitus (T2DM) in Mexican-Americans and Northern Europeans whereas these variations are not associated with T2DM in other populations. The aim of this study was to determine whether there is an association between specific CAPN10 diplotype (SNP-43, -19, and -63) and T2DM in the Korean population. Overall, 454 Korean patients with T2DM (male 230, female 224) and 236 non-diabetic controls (male 124, female 112) with no family history of diabetes were enrolled in this study. All the subjects were genotyped according to CAPN10 SNP-43, -19, and -63. The restriction fragment length polymorphism method was used for the three SNPs. There were eight estimated haplotype allelic variations. After adjusting for gender and age, the 111 haplotype was associated with a high risk of T2DM (P <0.0001). The 111/121 diplotype was associated with a high risk of T2DM (odds ratio =2.580, 95% confidence interval =1.602-4.155, P =0.001). The high-risk haplotype (112/121) in Mexican-Americans was not significant in our study population. In conclusion, we found that a novel 111/121 diplotype in Calpain-10 gene is associated with T2DM in the Korean population. PMID- 16721486 TI - A common functional exon polymorphism in the microsomal triglyceride transfer protein gene is associated with type 2 diabetes, impaired glucose metabolism and insulin levels. AB - The microsomal triglyceride transfer protein (MTP) is required for the assembly and secretion of apolipoprotein B-containing lipoproteins. Emerging evidence has indicated that the functional MTP exon polymorphism I128T is associated with dyslipidemia and other traits of the insulin-resistance syndrome, and the T128 variant seems to confer a reduced stability of MTP, resulting in reduced binding of LDL particles. The aim of the study was to elucidate the association of this MTP polymorphism with parameters of postprandial metabolism. A total of 716 male subjects from a postprandially characterized cohort (MICK) and a nested case control study (EPIC) of 190 incident type 2 diabetes cases and 380 sex- or age matched controls were genotyped for the I128T exon polymorphism. In comparison to homozygote subjects of the wild allele, carriers of the less common allele of the MTP T128 genotype showed significantly lower postprandial insulin levels (P=0.017), lower diastolic blood pressure (P=0.049) and had a lower prevalence of impaired glucose metabolism and diabetes type 2 (P=0.03) in the MICK. Consistent with this, we found a lower incidence of type 2 diabetes in male subjects of the nested case-control study in the T128 genotype (P=0.007). These results suggest that the rare allele of the MTP I128T polymorphism may be protective against impaired glucose tolerance, type 2 diabetes and other parameters of the metabolic syndrome. PMID- 16721487 TI - Ferroplasma cupricumulans sp. nov., a novel moderately thermophilic, acidophilic archaeon isolated from an industrial-scale chalcocite bioleach heap. AB - A new species of Archaea was isolated from an industrial mineral sulphide bioleach heap. Strain BH2, a non-motile pleomorphic coccus, was capable of chemomixotrophic growth on ferrous sulphate and yeast extract. Growth was not supported in the absence of yeast extract. Phylogenetic analysis based on the 16S rRNA gene showed that strain BH2 was most closely related to the species Ferroplasma acidiphilum; however, it showed only 95% sequence similarity with this species. Strain BH2 had a temperature optimum of 53.6 degrees C and a temperature range for growth between 22 and 63 degrees C. Thus, it is the first moderately thermophilic member of the genus Ferroplasma. The optimum pH for the growth of the strain occurred between pH 1.0 and 1.2 and the lowest pH at which growth was observed was 0.4. Based on 16S rRNA gene sequence analysis and other physiological characteristics, strain BH2 constitutes a new species within the genus Ferroplasma. The name Ferroplasma cupricumulans is proposed for the new species and strain BH2 (DSM 16651) is proposed as the type strain. PMID- 16721488 TI - Post-transplant lymphoproliferative disorder presenting as acute colonic perforation. PMID- 16721489 TI - Subretinal fibrosis and uveitis syndrome associated with ulcerative colitis. AB - BACKGROUND AND AIMS: We report a case of subretinal fibrosis and panuveitis syndrome as an extraintestinal manifestation of ulcerative colitis. MATERIALS AND METHODS: We present an interventional case report of a 40-year-old female patient with inactive ulcerative colitis referred to our center for refractory uveitis in her right eye. RESULTS/FINDINGS: She was diagnosed with subretinal fibrosis and uveitis syndrome after work-up ruled out tuberculosis, bartonellosis, Lyme disease, and sarcoidosis. Right eye multifocal choroiditis and panuveitis syndrome with subretinal fibrosis was diagnosed, and posterior pole subretinal fibrosis caused a 90% loss of visual efficiency. She had concurrent dermatological manifestations but no gastrointestinal symptoms. One year of immunosuppressive therapy with methotrexate (7.5 mg/week) controlled ocular inflammation. Two colitis episodes, with no visual involvement, were recorded during follow-up. Her final, right-eye, best-corrected visual acuity remained stable at 0.160 due to scarring sequela after 7 years of follow-up. INTERPRETATION/CONCLUSION: Subretinal fibrosis and uveitis syndrome can occur as a sight-threatening extraintestinal manifestation of ulcerative colitis. PMID- 16721491 TI - Reply to the report: epidemiology of rheumatoid arthritis in Turkey (by N. Akkoc, S. Akar). PMID- 16721490 TI - Detection of micrometastases and skip metastases with ex vivo sentinel node mapping in carcinoma of the colon and rectum. AB - BACKGROUND: The debate over sentinel lymph node mapping (SLNM) and focused pathologic examination to detect micrometastases in patients with colorectal cancer (CRC) continues. We present in this paper our experience with SLNM for CRCs to improve staging. In addition, we have detailed the mapping procedure on an anatomical basis to define skip metastasis. MATERIALS AND METHODS: Forty-seven patients underwent ex vivo SLNM. Immediately after resection, 1 ml of patent blue VF was injected submucosally around the tumor. Lymph nodes harvested from the first 15 patients were mapped in a standard fashion as the blue-stained nodes (SLNs), and the others (non-SLNs) were dissected away. In the remaining 32 patients, the lymph nodes were also mapped separately in relation to their anatomic location and described as epicolic-paracolic, intermediate, and principal. The blue-stained nodes (SLNs) and non-SLNs, negative by hematoxylin and eosin stain, were further stained with cytokeratin immunohistochemical analysis and carcinoembryonic antigen. RESULTS: A total of 873 histologically confirmed LNs were examined with a mean of 18.6+/-8.1 nodes per patient. In 46 of 47 patients (97.8%), SLNs were identified. Immunohistochemical staining revealed micrometastases in the lymph nodes of four patients, which were negative by conventional methods. Anatomical skip metastases were noted in 4 of 32 patients studied (12.5%). CONCLUSION: Ex vivo SLNM in CRCs is a feasible technique with a high SLN identification rate. Results of anatomical mapping of lymph nodes correlates with the limited literature, suggesting that occult skip metastases can occur in the apical lymph node group and may occur outside the resected area. PMID- 16721492 TI - Hypercoagulability: interaction between inflammation and coagulation in familial Mediterranean fever. AB - Familial Mediterranean fever (FMF) patients in clinical remission are reported to have increased baseline inflammation. Normal function of the natural anticoagulant pathways is particularly needed in diminishing inflammatory responses. In the presence of subclinical inflammation, natural anticoagulant response may be exaggerated. We aimed to observe the anticoagulant-procoagulant status in attack-free FMF patients. Twenty-seven FMF patients diagnosed in accordance with Tel-Hashomer criteria, and 26 healthy controls were included. All patients were attack-free under regular colchicine treatment. Amyloidosis, autoimmunity, accompanying liver and renal disease, and vasculitis were excluded. Predisposing factors for thrombosis were not present. Acute phase reactants (APRs), anticardiolipin antibody positivity, prothrombin time (PT), activated prothrombin time, thrombin time (TT) and d-dimer, protein C activity, activated protein C resistance, free protein S, antithrombin, lupus anticoagulant, human prothrombin fragment F 1 + 2, and human thrombin/antithrombin III complex were analyzed for all subjects. APRs were comparable with controls. Autoimmune markers were negative in all. Anti-streptolysin titers were significantly different than the control group. PT, TT, protein C activity, and F 1 + 2 levels were significantly different from those of healthy controls. Shortened PT and TT, decreased protein C activity vs increased levels of F 1 + 2 suggested a hypercoagulable state in our patients. The hypercoagulable state detected in FMF patients suggests that screening with abnormal coagulation tests may be beneficial for tracing the future consequences of subclinical inflammation in these patients. Studies covering larger groups of patients are needed to verify the currently observed hypercoagulable status in FMF. PMID- 16721493 TI - High titres of IgM-antiphospholipid antibodies are unrelated to pathogenicity in patients with non-Hodgkin's lymphoma. AB - Heterogeneity in the mechanisms of coagulation may contribute to an increased thrombotic risk for patients with malignancies. The coincidence of malignancies and antiphospholipid antibodies (aPL) have been described in several important epidemiological studies. The pathological significance of aPL in patients with malignancies is, however, still unclear. In this study, we investigated the clinical manifestations of four patients with elevated IgM-aPL titres lying outside the region signifying 95% of normal cases and with a history of non Hodgkin's lymphoma. The patients had elevated IgG- and IgM-anticardiolipin antibodies (aCL) and also tested positive for lupus anticoagulants. Other aPL were measured, and we found high positive results for all tested antibodies in three patients. The production of aPL, however, occurred in the absence of thrombotic complications. No thromboembolic manifestations occurred during the follow-up period either. It could also be demonstrated that the degree to which the aCL titre was elevated resembles the elevation of the non-classical antiphospholipid antibodies, but not that of beta2-GP-1 or anti-annexin antibodies. Therefore, it can be postulated that these extremely high levels of IgM-aCL antibodies do not enhance the risk of thrombosis and may be completely different from aCL antibodies in an antiphospholipid syndrome patient population without malignancies. In particular, haematological and lymphoproliferative malignancies may indeed be associated with the generation of aPL, but do not necessarily enhance the thrombophilic risk in these patients. PMID- 16721494 TI - Vasculitis in siblings with familial Mediterranean fever: a report of three cases and review of the literature. AB - Familial Mediterranean Fever (FMF) is characterized by recurrent attacks of self limited polyserositis and fever. Several types of vasculitis are associated with FMF: polyarteritis nodosa, Henoch-Schonlein purpura (HSP), and protracted febrile myalgia (PFM). We describe three cases of vasculitis in four siblings of a Sephardic Jewish family with FMF and reviewed the literature. One brother and one sister developed severe HSP with intestinal involvement while another brother developed PFM. Genetic tests in three brothers confirmed the M694V mutation on both alleles. Vasculitides may be a clinical feature of FMF with a higher familiar prevalence. MEFV mutations may act as a genetic susceptibility factor for vasculitides in FMF patients. PMID- 16721495 TI - A toddler with recurrent oral and genital ulcers. AB - In western countries, when a child presents with recurrent oral ulcers and colitis, the diagnosis of Crohn's disease is mostly made. In our patient, the diagnosis was Behcet's disease with gastrointestinal manifestations. Behcet's disease with gastrointestinal manifestations has a similar clinical presentation to Crohn's disease, but there is more organ involvement and the prognosis is more severe in the former. Because there is limited experience in the treatment of Behcet's disease in the paediatric population, successful and unsuccessful treatment modalities in both paediatric and adult populations should be reported. PMID- 16721496 TI - Severe soft tissue infection of the thigh after vaginal erosion of transobturator tape for stress urinary incontinence. AB - Since the beginning of use of synthetic midurethral slings, several complications, usually benign, have been reported. Recently, three consecutive cases of severe thigh infection secondary to transobturator insertion of a synthetic tape alarmed us. This is a case report about these three cases and a review of literature about complications of transobturator tapes. PMID- 16721500 TI - Impaired working speed and executive functions as frontal lobe dysfunctions in young first-degree relatives of schizophrenic patients. AB - The aim of the investigation was to detect neuropsychological markers, such as sustained and selective attention and executive functions, which contribute to the vulnerability to schizophrenia especially in young persons. Performance was assessed in 32 siblings and children of schizophrenic patients and 32 matched controls using Wisconsin Card Sorting Test, Colour-Word-Interference-Test, Trail Making Test, and d2-Concentration-Test. The first-degree relatives showed certain impairments on all four tests, in particular, slower times on all time-limited tests. These results suggest the need for more time when completing neuropsychological tasks involving selected and focused attention, as well as cognitive flexibility, as a possible indicator of genetic vulnerability to schizophrenia. PMID- 16721501 TI - Depression and activity limitations: examining gender differences in the general population. AB - BACKGROUND: This study examines the relationship between major depression, subclinical depressive symptomatology and activity limitation in a representative sample of Canadian men and women, with a particular emphasis on examining gender differences. METHODS: Over a 4-year period, the three waves of data were collected from a representative sample of Canadian adults aged 18-60 beginning in 1994. A brief diagnostic interview for major depression, current limitations in home, work, and other activities (e.g., leisure), presence of each of 13 chronic medical conditions, and other sociodemographic information were gathered. Regression analyses adjusted for potential confounding of time-dependent and time invariant covariates. These analyses were performed separately for men and women. RESULTS: Major depression had a significant adverse impact on all types of activities for the entire community sample, and this impact was generally robust when controlling for potential confounders. Gender differences were observed for leisure activities with depressed men showing a higher likelihood of limitation in these activities than depressed women. CONCLUSIONS: The findings provide further evidence that major depression leads to impairments in a range of daily activities. Gender differences in the impact of depression on leisure activities may be important to consider in depression treatment. PMID- 16721502 TI - Subjective quality of life of patients with obsessive-compulsive disorder. AB - BACKGROUND: Obsessive-compulsive disorder (OCD) is the fourth most frequent diagnosis in psychiatry. Patients with OCD suffer from obsessive thinking and compulsive behavior, which impact their everyday life negatively. OBJECTIVE: Subjective quality of life (QoL) in patients with OCD was examined and compared to the general German population and to patients with schizophrenia. METHODS: Seventy-five patients, aged 21-72, with OCD (ICD 10 F42.0-F42.2) were recruited from the outpatient clinic for anxiety disorders at the Department of Psychiatry of the University of Leipzig. By means of the WHOQOL-BREF, QoL was assessed in patients with OCD in a representative sample of patients with schizophrenia and in a sample of the general population of Saxony/Germany. RESULTS: Compared with the general population, QoL in patients with OCDs was lower in all domains of the WHOQOL-BREF. Unexpectedly, QoL in patients with OCDs was lower in the domains "psychological well-being" and "social relationships" when compared with schizophrenia patients. CONCLUSIONS: OCD has a substantial adverse effect on patients' subjective QoL, which may be even greater than the adverse effect of schizophrenia. Therefore, it will be necessary to include interventions in the treatment of OCD aimed at improving residual deficits in psychosocial functioning and QoL. PMID- 16721503 TI - Dramatic stiffening of ultrathin polymer films in the rubbery regime. AB - Recently, we (P.A. O'Connell, G.B. McKenna, Science 307, 1760 (2005)) introduced a novel nano-bubble inflation method to measure the absolute creep compliance of nanometer thick polymer films. In that work it was shown that even at film thicknesses as small as 27.5nm the glass temperature was unchanged for poly(vinyl acetate) (PVAc). Perhaps more importantly, and the subject of the present work, was the observation that these ultrathin films show a dramatic stiffening in the rubbery plateau regime, i.e., the compliance was reduced by over two orders of magnitude compared to the bulk material. In the present work we substantiate the previous results in a study of the thickness dependence of the rubbery compliance of PVAc and polystyrene (PS) films for thicknesses from 13nm to 276nm. We show the substantial stiffening of the plateau region for both materials. Furthermore, the rubbery compliance (inverse of stiffness) scales with approximately the second power ( 1.8+/-0.2) in the film thickness for both materials. PMID- 16721504 TI - Risk factors related to recurrence in inguinal hernia repair: a retrospective analysis. AB - The aim of this study was to analyze and evaluate the long-term recurrence rate and risk factors for inguinal hernia recurrence in patients treated by the Shouldice suture repair. A total of 293 hernias treated by Shouldice suture technique in 1992 were studied retrospectively. After a 10-year follow-up, 15 potential risk factors for recurrence were assessed in 142 patients undergoing 171 Shouldice repairs. Recurrent hernias showed a significantly higher (22.0%) recurrence rate than primary inguinal hernias (7.7%). Furthermore, an age of more than 50 years, smoking, and the presence of two or more similarly affected relatives were found to be independent risk factors for recurrence. The present study underlines the importance of patient-related risk factors for the development of a recurrent inguinal hernia. Patients at risk should preoperatively be identified in order to improve treatment by, for example, the application of mesh techniques. PMID- 16721505 TI - The role of HIV serostatus disclosure in antiretroviral medication adherence. AB - This study examined the relationship between HIV serostatus disclosure and adherence to antiretroviral therapy (ART). The study was conducted with 215 HIV seropositive patients who demonstrated poor adherence (<80%) and who were in serodiscordant relationships. Participants completed self-report measures regarding HIV serostatus disclosure and reasons for missing ART doses, as well as electronic monitoring of ART adherence (MEMS caps). Overall, 19% of the sample reported missing medication doses in the last two months due to concerns regarding serostatus disclosure. Participants who reported greater serostatus disclosure to others demonstrated higher rates of adherence, and this relationship remained after controlling for other explanatory variables. The relationship between disclosure and adherence was not mediated by practical support for adherence from others. Interventions to improve ART adherence should address the role of serostatus disclosure by providing patients with skills to maintain adherence in contexts of non-disclosure and to make informed choices regarding selective disclosure. PMID- 16721506 TI - Accuracy of audio computer-assisted self-interviewing (ACASI) and self administered questionnaires for the assessment of sexual behavior. AB - This study examined the accuracy of two retrospective methods and assessment intervals for recall of sexual behavior and assessed predictors of recall accuracy. Using a 2 [mode: audio-computer assisted self-interview (ACASI) vs. self-administered questionnaire (SAQ)] by 2 (frequency: monthly vs. quarterly) design, young women (N =102) were randomly assigned to one of four conditions. Participants completed baseline measures, monitored their behavior with a daily diary, and returned monthly (or quarterly) for assessments. A mixed pattern of accuracy between the four assessment methods was identified. Monthly assessments yielded more accurate recall for protected and unprotected vaginal sex but quarterly assessments yielded more accurate recall for unprotected oral sex. Mode differences were not strong, and hypothesized predictors of accuracy tended not to be associated with recall accuracy. Choice of assessment mode and frequency should be based upon the research question(s), population, resources, and context in which data collection will occur. PMID- 16721508 TI - Postoperative radiotherapy after radical prostatectomy for prostate carcinoma: the experience of the Brescia Radium Institute. AB - PURPOSE: The purpose of this study was to evaluate the efficacy of postoperative radiotherapy in reducing the incidence of prostate carcinoma (PCa) recurrences after radical prostatectomy (RP), define the importance of the time interval between surgery and radiotherapy for prognosis and the toxicity of the treatment in comparison with radiotherapy or surgery alone. MATERIALS AND METHODS: We examined 97 patients who consecutively underwent postoperative radiotherapy after RP between 1980 and 2003. The treatment was considered "adjuvant" if was conducted less than 6 months after RP, if there was no macroscopic residual disease and if there was no progressive increase in serum prostate-specific antigen (PSA) and "salvage" if performed more than 6 months after RP, for the presence of macroscopic recurrence or with rising PSA. Radiotherapy was salvage in 56 patients and adjuvant in 41. Age range was 60-70 years in 80% of patients, and the Karnofsky index was over 80 in 78% of cases. Histology revealed extracapsular spread in 60% of patients. Preradiotherapy PSA was higher than 1 ng/ml in 36%. Radiotherapy was performed on the surgical bed only in 80%, and the total dose was 70 Gy in 62% of cases. RESULTS: Recurrence-free survival (RFS) at 5 years and 10 years was 53+/-8% and 32+/-14.2%, respectively, for the whole sample; 76+/-9% and 38+/-2.7% for patients treated with adjuvant radiotherapy and 36+/-10% and 28+/-10% for those treated with salvage radiotherapy (p<0.01). Moreover, the 5-year RFS was better in the group treated with adjuvant radiotherapy and PSA less than or equal to 1 ng/ml (p<0.05). Treatment toxicity was acceptable. CONCLUSIONS: Postoperative radiotherapy improves RFS and reduces the risk of local recurrence. The best results are obtained with early postoperative treatment ("adjuvant"); adjuvant radiotherapy of high-risk forms yields better results if performed with PSA less than or equal to 1 ng/ml. PMID- 16721509 TI - Risk management in radiotherapy: analysis of insurance claims. AB - PURPOSE: The purpose of this study was to analyse risk factors in radiotherapy with regard to those where protocols have been implemented for their prevention and those that are known but still present, thus requiring attention and remedial measures. MATERIALS AND METHODS: To demonstrate the existence of risk factors that have not been eliminated and that cause damage in radiotherapy practice, we examined insurance claims related to such radiotherapy procedures. Analysis of the claims made over a 10-year period was performed on the basis of radiotherapists (256 in 2004) insured with the professional association. The total number of claims in the 10-year period was 26. RESULTS: The type of claim was examined in each of the 26 cases, with as many as 15 being related to radiotherapy injury to tissues surrounding the neoplasm, thus confirming this to be the most frequent cause for insurance claims. The other 11 causes are also reported. CONCLUSIONS: Now that the historical causes of error that led to incorrect radiotherapy treatment have been eliminated through strict adherence to protocols, risk management in radiotherapy identifies side effects as the main source of risk for patients and physicians. Data analysis confirms the need to implement risk management procedures in radiotherapy in which insurance claims (15/26 cases) are motivated by side effects of treatment. PMID- 16721512 TI - Taxon-specific suffixes for vernacular names. PMID- 16721513 TI - P-glycoprotein and 'lipid rafts': some ambiguous mutual relationships (floating on them, building them or meeting them by chance?). AB - P-glycoprotein (P-gp) is an active membrane transporter responsible for cell detoxification against numerous amphiphilic compounds, leading to multidrug resistance in tumor cells. It displays entangled connections with its membrane environment since it recognizes its substrates within the cytosolic leaflet and it also translocates some endogenous lipids to the exoplasmic leaflet. Regarding its relationships with membrane microdomains, 'lipid rafts', a literature analysis concludes that (i) P-gp also exists in rafts and non-raft membrane domains, depending on the cell considered, the experimental conditions and the method used to test it; (ii) cholesterol has a positive influence on P-gp function, and this may be a direct effect of the free cholesterol present in membrane or an indirect effect mediated by the cholesterol-enriched microdomains; (iii) when present in rafts, P-gp interacts with protein partners regulating its activity; (iv) P-gp is a lipid translocase that handles the raft-constituting lipids with particular efficiency, and it also influences membrane trafficking in the cell. PMID- 16721520 TI - Rectal volume tolerability and anal pressures in patients with fecal incontinence treated with sacral nerve stimulation. AB - PURPOSE: Sacral nerve stimulation has proven to be a promising treatment for fecal incontinence when conventional treatment modalities have failed. There have been several hypotheses concerning the mode of action of sacral nerve stimulation, but the mechanism is still unclear. This study was designed to evaluate the results of rectal volume tolerability, rectal pressure-volume curves, and anal pressures before and six months after permanent sacral nerve stimulation and to investigate the mode of action of sacral nerve stimulation. METHODS: Twenty-nine patients with incontinence (male/female ratio = 6/23; median age, 58 (range, 29-79) years) underwent implantation of a permanent sacral electrode and neurostimulator after a positive percutaneous nerve evaluation test. Wexner incontinence score, rectal distention with thresholds for "first sensation," "desire to defecate," and "maximal tolerable volume," rectal pressure volume curves, anal resting pressure, and maximum squeeze pressure were evaluated at baseline and at six months follow-up. RESULTS: Median Wexner incontinence score decreased from 16 (range, 6-20) to 4 (range, 0-12; P < 0. 0001). Median "first sensation" increased from 43 (range, 16-230) ml to 62 (range, 4-186) ml (P = 0.1), median "desire to defecate" from 70 (range, 30-443) ml to 98 (range, 30 327) ml (P = 0.011), and median "maximal tolerable volume" from 130 (range, 68 667) ml to 166 (range, 74-578) ml (P = 0.031). Rectal pressure-volume curves showed a significant increase in rectal capacity (P < 0.0001). The anal resting pressure increased significantly from 31 (range, 0-109) cm H(2)O to 38 (range, 0 111) cm H(2)O (P = 0.045). No significant increase in maximum squeeze pressure was observed. CONCLUSIONS: For patients with fecal incontinence successfully treated with sacral nerve stimulation, there was a significant increase in rectal volume tolerability and rectal capacity. A significant increase in anal resting pressure, but not in maximum squeeze pressure, was found. We suggest that sacral nerve stimulation causes neuromodulation at spinal level. PMID- 16721522 TI - Cervical spine disorders in farm workers requiring neck extension actions. AB - BACKGROUND: Cervical extension is considered a risk factor for provoking radicular and spinal cord compromise. However, there have been no reports on the relation between extension strain (ES) and cervical spine disorders: degenerative cervical spine disorders, dynamic canal stenosis (DCS), and cervical myelopathy. We performed a cross-sectional study to investigate the relation. METHODS: Orthopedic examinations were performed on 177 grape-growers (ES-exposed group) and 191 eggplant-growers (control group) between May and August 2000; and patients with degenerative cervical spine disorders, DCS, and cervical myelopathy were identified in the two groups. ES, degenerative cervical spine disorders, DCS, and cervical myelopathy were regarded as exposure variables; and age, sex, and working years were regarded as confounders. In cases where the subject was exposed to each exposure variable, multivariate-adjusted odds ratios to degenerative cervical spine disorders, DCS, and cervical myelopathy and multivariate-adjusted odds ratios regarding cervical myelopathy as the dependent variable were calculated by unconditional logistic regression analysis. RESULTS: Multivariate unconditional logistic regression analysis showed a significant odds ratio of ES to degenerative cervical spine disorders [2.72, 95% confidence interval (CI) 1.62-4.56]. Multivariate unconditional logistic regression analysis showed significant odds ratios for DCS (4.50, 95% CI 2.03-9.96) and age (1.07, 95% CI 1.01-1.14) regarding cervical myelopathy as the dependent variable. CONCLUSIONS: These findings suggested that ES of the cervical spine is a risk factor for degenerative cervical spine disorders, and DCS and the aging process are risk factors for cervical myelopathy. PMID- 16721523 TI - Serum interleukin-6 response after spinal surgery: estimation of surgical magnitude. AB - BACKGROUND: Serum interleukin-6 (IL-6) has been used for quantitative estimation of the surgical magnitude of major cardiac and thoracoabdominal surgery, but there have been few studies assessing IL-6 as a marker of surgical magnitude of spinal surgery. METHODS: We investigated the changes in IL-6 response in comparison to other parameters of surgical magnitude and spinal surgery procedures. The study included 40 patients electively undergoing spinal surgery. The patients were divided into four groups: lumbar laminectomy with posterolateral fusion (PLF), lumbar laminotomy, lumbar open discectomy, and cervical laminoplasty. Serum IL-6, C-reactive protein (CRP), creatine kinase (CK), and the white blood cell (WBC) count were determined in venous blood before surgery, at the end of surgery, and 6 h and the first, third, and seventh days after surgery. RESULTS: Serum IL-6 peaked on the first day and returned to a normal value by the seventh day. The peak IL-6 concentrations on the first day after surgery significantly correlated with CRP, CK, duration of surgery, and estimated blood loss. Regarding lumbar surgeries, the peak IL-6 for laminectomy/PLF was significantly higher than that for laminotomy/open discectomy or for cervical laminoplasty. CONCLUSIONS: Serum IL-6 on the first day varied depending on the surgical procedure used. Therefore, it might be a quantitative marker of surgical magnitude following spinal surgery. PMID- 16721524 TI - Validation of the Japanese Society for Surgery of the Hand Version of the Quick Disability of the Arm, Shoulder, and Hand (QuickDASH-JSSH) questionnaire. AB - BACKGROUND: The Quick Disability of the Arm, Shoulder, and Hand (QuickDASH) questionnaire is a region-specific, self-administered questionnaire, which consists of a disability/symptom (QuickDASH-DS) scale, and the same two optional modules, the work (DASH-W) and the sport/music (DASH-SM) modules, as the DASH. After the Japanese version of DASH (DASH-JSSH) was cross-culturally adapted and developed, we made the Japanese version of QuickDASH (QuickDASH-JSSH) by extracting 11 out of 30 items of the DASH-JSSH regarding disability/symptoms. The purpose of this study was to test the reliability, validity, and responsiveness of QuickDASH-JSSH. METHODS: A series of 72 patients with upper extremity disorders completed the QuickDASH-JSSH, the 36-Item Short-Form Health Survey (SF 36), and the Visual Analog Scale (VAS) for pain. Thirty-eight of the patients were reassessed for test-retest reliability 1 or 2 weeks later. Reliability was investigated by the reproducibility and internal consistency. To analyze the validity, a principal component analysis and the correlation coefficients between the QuickDASH-JSSH and the SF-36 were obtained. The responsiveness was examined by calculating the standardized response mean (SRM; mean change/SD) and effect size (mean change/SD of baseline value) after carpal tunnel release of the 17 patients with carpal tunnel syndrome. RESULTS: Cronbach's alpha coefficient in the QuickDASH-DS was 0.88. The intraclass correlation coefficient (ICC) for the same was 0.82. The unidimensionality of the QuickDASH-DS was confirmed. The correlation coefficients between the QuickDASH-DS and the DASH-DS, DASH-W, or the DASH-SM were 0.92, 0.81, or 0.76, respectively. The correlation coefficients between the QuickDASH-DS score and the subscales of the SF-36 ranged from -0.29 to -0.73. The correlation coefficient between the QuickDASH-DS score and the VAS for pain was 0.52. The SRM/effect size of QuickDASH-DS was -0.54/-0.37, which indicated moderate sensitivity. CONCLUSION: The Japanese version of QuickDASH has equivalent evaluation capacities to the original QuickDASH. PMID- 16721525 TI - Method to reduce variations of inclination angle of the acetabular component during mini-incision hip arthroplasty. AB - BACKGROUND: To reduce variations of cup inclination after total hip arthroplasty using the mini-incision posterior approach, we introduced two techniques, one at a time. The first technique is measuring a pelvic tilt angle in the frontal plane in the initial lateral position in the operating room. The second technique is using a tilt-meter to adjust the direction of a cup holder. The purpose of this study was to evaluate the usefulness of these techniques. METHODS: For 106 hips operated on, the cementless acetabular component was impacted using a cup holder targeting 45 degrees in inclination and 20 degrees in anteversion. These hips were divided into three consecutive groups. For the first group (30 hips), no radiograph in the lateral position was obtained, and the alignment frame of the cup holder was aligned parallel to the floor by eye measurement. For the second group (56 hips), we measured the pelvic tilt angle, and tilted the alignment frame by eye measurement. For the third group (20 hips), we measured the pelvic tilt angle and tilted the alignment frame using the tilt meter. Inclination and anteversion angles were measured on postoperative radiographs. The absolute value of the difference between the measured angle and the target angle was defined as the inclination error or anteversion error, respectively. RESULTS: The inclination error was more than 5 degrees for 33% in the first group, 20% in the second group, and 0% in the third group (P = 0.015, chi-squared test). There was a significant difference between the first group and the third group (P = 0.0039). For the anteversion error, there were no significant differences among the three groups. CONCLUSIONS: Adjustment using a tilt-meter after measuring a pelvic tilt angle is a useful method to reduce the rate of large inclination error. PMID- 16721526 TI - Scoliosis in cat cry syndrome. AB - BACKGROUND: Cat cry syndrome is an autosomal disease accompanying abnormal deletion of chromosome 5 and occurs in only 1 of 50,000 neonates. Scoliosis has been reported as a skeletal complication in cat cry syndrome. The characteristics and causes of scoliosis in this rare syndrome are unknown. The purpose of this study was to present the characteristics of scoliosis in cat cry syndrome and to speculate on its causative mechanisms. METHODS: We report on 11 cases (5 boys and 6 girls) of cat cry syndrome. Detailed investigations of scoliosis, as well as physical and imaging examinations, were performed to characterize scoliosis and its causes. Average age at initial diagnosis of scoliosis was 4.3 years, and average age at final examination was 11.8 years. RESULTS: The incidence of scoliosis was as high as 73% (8/11). Most cases show a single right thoracic curve. Of the 8 patients with scoliosis, 3 patients who had increased muscular tone showed marked progression of scoliosis with growth. CONCLUSIONS: Muscular hypertonia may play a key role in the progression of scoliosis in cat cry syndrome. PMID- 16721527 TI - Asymmetry of mediolateral laxity of the normal knee. AB - BACKGROUND: Understanding the normal kinematics of the joints is important for reconstructive surgery. However, only a few extensive studies have been done on medial and lateral laxity of the normal knee. METHODS: Radiographs of 50 normal knees were obtained under varus and valgus stress in both extension and flexion and the relative angle of the articular surface was measured. RESULTS: In extension, the mean angle was 4.9 degrees in varus stress and 2.4 degrees in valgus stress. In flexion, the mean angle was 4.8 degrees in varus stress and 1.7 degrees in valgus stress. Lateral laxity was significantly greater than medial laxity in both extension and flexion. CONCLUSIONS: Lateral laxity may be necessary for the medial pivot movement of the normal knee. There is some disagreement regarding the importance of pursuing the perfect rectangular gaps during total knee arthroplasty (TKA). The methods for measuring the tension of soft tissues during the operation are not accurate and do not always reflect the postoperative tensions of dynamic phases, such as walking and standing. Slight lateral laxity can be accepted with TKA, and further studies are necessary to determine whether prosthesis lift-off occurs in the replaced knee with slight lateral laxity similar to that in the normal knee. PMID- 16721528 TI - Skin behavior during leg lengthening in patients with achondroplasia and hypochondroplasia: a short-term observation during leg lengthening. AB - BACKGROUND: Although distraction osteogenesis is a recently established method of limb lengthening, it is not well known how soft tissues behave during bone lengthening. Because skin is tissue that can be easily and directly observed, we analyzed skin extension during limb lengthening to study the condition of soft tissues. METHOD: We examined three achondroplastic and two hypochondroplastic patients who underwent bilateral tibial lengthening with unilateral bone lengtheners. To analyze skin extension during leg lengthening, the bilateral skin surface was marked in a latticework pattern, starting 10 cm proximal to the knee joint down to 10 cm distal to the ankle joint, with black ink. The longitudinal length and the area of lattices, and the circumferential length of legs were measured. RESULTS: We found that: (1) the longitudinal extension of the skin during leg lengthening occurs not only in the bone lengthening portion between fixator pins but also around the knee or ankle joints; (2) circumferential length of legs was decreased, and soft tissue volume also does not increase in proportion to the increase in leg length. Soft tissue volume started to increase in the central region of the legs after a 30% increase in leg length was obtained, resulting in the 10% increase in the volume at the end point. CONCLUSIONS: These data suggest that soft tissue extension precedes soft tissue neogenesis at least during the initial period of leg lengthening, possibly based on its viscoelastic properties. After a 30% increase in leg length is obtained, soft tissue neogenesis takes place in the central region of the leg. PMID- 16721529 TI - Effects of ligament repair on laxity and creep behavior of an early healing ligament scar. AB - BACKGROUND: Previous clinical studies have reported that conservatively managed medial collateral ligament (MCL) injuries remained unstable 9 years post injury with subjective complaints of muscle weakness, reinjuries, and post-traumatic osteoarthritis. Animal studies have also reported that healing MCLs were weaker and more lax than controls. Therefore, our purpose was to study the early effects of ligament repair on scar laxity, creep, and creep recovery in a rabbit model of bilateral medial collateral ligament (MCL) injury. METHODS: Each rabbit had one MCL cut in midsubstance which was not repaired, while the other MCL had a sagittal Z-plasty repair. Six weeks after surgery, isolated bone-MCL-bone complexes were biomechanically tested for MCL laxity, cyclic creep, creep recovery, and then loaded to ultimate failure. RESULTS: Nonrepaired scars were significantly more lax than both repaired Z-plasty scars and normal controls. In contrast, there was no significant difference in MCL laxity between repaired scars and normal controls. There were no significant differences between nonrepaired or repaired scars for either cyclic creep and creep recovery. Both crept significantly more and recovered significantly less than normal controls after the same load history. There were no significant differences in the failure load, stiffness, and tensile strength between gap scars and Z-plasty scars. All healing ligaments had significantly lower failure load, stiffness, and tensile strength than normal controls. CONCLUSIONS: The present study demonstrates that ligament repair is effective in decreasing short-term laxity of ligament scars. These 6-week scars would be prone to abnormal creep if loaded excessively. PMID- 16721530 TI - Biomechanical comparison of different fixation methods on the tibial side in anterior cruciate ligament reconstruction: a biomechanical study in porcine tibial bone. AB - BACKGROUND: The fixation strength of the hamstring tendon graft on the tibial side is considered the weak point in anterior cruciate ligament (ACL) reconstruction. The purpose of this study was to compare the biomechanical characteristics of four types of ACL reconstruction methods on the tibial side. METHODS: Extensor digitorum tendons harvested from fresh bovine forelimbs were used as the graft materials in this study. Twenty-eight porcine tibias were divided into four groups based on different fixation methods. Group D was fixed using a double spike plate (DSP), group I was fixed using an interference screw, group DI-80 was fixed using both an interference screw and DSP (80 N tension was applied to DSP), and group DI-150 was fixed using both an interference screw and DSP (150 N tension was applied to DSP). RESULTS: The displacement of the grafts in response to 500 cycles of 0-150 N loading was significantly greater in groups D (10.3 +/- 15 mm) and I (5.5 +/- 1.7 mm) than that in groups DI-80 (2.1 +/- 0.3 mm) and DI-150 (1.2 +/- 0.4 mm), with no significant differences between groups DI-80 and DI-150. The ultimate failure load and stiffness were significantly higher in groups DI-80 (745 +/- 156 N, 103 +/- 17 N/mm) and DI-150 (801 +/- 129 N, 151 +/- 35 N/mm) than those in groups D (374 +/- 53 N, 70 +/- 13 N/mm), and I (520 +/- 76 N, 78 +/- 18 N/mm). The stiffness of group DI-150 was significantly greater than that of group DI-80. CONCLUSIONS: Our results indicate that the initial fixation strength of the hamstring tendon can be increased by using an interference screw combined with DSP on the tibial side. PMID- 16721531 TI - In vivo banking for vascularized autograft bone by intramuscular inoculation of recombinant human bone morphogenetic protein-2 and beta-tricalcium phosphate. AB - BACKGROUND: The gold standard of bone graft substitutes is vascularized autologous bone. Our strategy to acquire this bone substitute is using recombinant human bone morphogenetic protein (rhBMP)-2 with beta-tricalcium phosphate (TCP) as carrier to induce bone tissue in muscle and grafting as a muscle-pedicled bone flap. In our previous studies, morphologically controlled living bone was successfully induced in a rat muscular pedicle. Moreover, concurrent administration of bisphosphonate controlled the subsequent resorption of the induced bone. This study was conducted (1) to confirm that when the size of carrier is increased, the induced bone can form the same in the center as well as on the periphery of the carrier, and (2) to simulate in vivo banking for vascularized autograft bone. METHODS: In experiment 1200 microg rhBMP-2 and 10( 5) M bisphosphonate (Minodronate) were applied to a 1-cm-diameter sphere of beta TCP. This sphere was then inoculated into rabbit quadriceps and harvested 5 weeks later (n = 3). In experiment 2, beta-TCP was generated as a one-third hollow cylinder with 4-mm outer radius, 2-mm inner radius, and 10-mm length. After 50 microg rhBMP-2 and 10(-5) M Minodronate were applied, this structure was then inoculated into rat quadriceps. Four weeks later, the induced bone was transplanted as a muscle-pedicled flap onto the adjacent femur and harvested after 4 and 8 weeks more (n = 3). RESULTS: In experiment 1, good bone formation was observed at the central as well as the peripheral region of the sphere. In experiment 2, good bone formation and bony union to the femur at both 4 and 8 weeks were detected on the X-ray film. The presence of new bone formation around the screw was observed. CONCLUSIONS: Transplantation of the beta-TCP carrier treated with rhBMP-2 and bisphosphonate into the muscular tissue may have clinical potential for in vivo banking for muscle-pedicled bone flaps. PMID- 16721532 TI - Calcium phosphate cement in wrist arthrodesis: three cases. AB - BACKGROUND: With degenerative pathologies, arthrodesis of the wrist is a procedure reserved for preservative interventions that have failed. The least invasive procedure is Gill's technique, as modified by Luboshitz. To improve the results, we proposed a study of the advantages of local application of calcium phosphate cement. METHODS: The procedure consisted of refreshing the remaining articular surfaces, obtaining a dorsal corticocancellous graft from the distal radius, and replacing the bone substance lost when removing the graft with calcium phosphate cement followed by smoothing flat the arthrodesis area to limit tendon conflicts. RESULTS: In our three cases with an average follow-up of 23 months, no complications were noted and radiological consolidation was achieved in 2 months. CONCLUSIONS: In view of this short series, we think that the addition of calcium phosphate cement to Gill's technique as modified by Luboshitz should improve the results of arthrodesis of degenerative wrists. PMID- 16721533 TI - Vertebral hemangioma compressing the thoracic spinal cord: application of computer-aided navigation and intraoperative spinal sonography for surgery through anterior and posterior approaches. PMID- 16721534 TI - En bloc resection, extracorporeal irradiation, and reimplantation of an entire tibia. PMID- 16721535 TI - Synovial impingement after posterior cruciate-retaining total knee arthroplasty for rheumatoid arthritis. PMID- 16721536 TI - Fracture of the precoated femoral component after cemented total hip arthroplasty. PMID- 16721537 TI - Giant cell-rich tumor of the rib. PMID- 16721538 TI - What I have learned about the ACL: utilizing a progressive rehabilitation scheme to achieve total knee symmetry after anterior cruciate ligament reconstruction. AB - Anterior cruciate ligament surgery and rehabilitation have changed drastically during the past 30 years. The patellar tendon autograft fixed with buttons provides tight bone-to-bone placement of the graft and quick bony healing, which allows accelerated rehabilitation to obtain full range of motion and strength. Although surgical stability is easily reproducible, long-term patient satisfaction is difficult to guarantee. Full knee range of motion should be compared to that of the contralateral normal knee, including full hyperextension. We followed the progress of all patients to gauge the utility of our rehabilitation program. In order of importance, the lack of normal knee range of motion (within 2 degrees extension and 5 degrees of flexion compared with that of the normal knee), partial or total medial meniscectomy, partial or total lateral meniscectomy, and articular cartilage damage were related to lower subjective scores. Rehabilitation after ACL reconstruction must first strive to achieve full symmetrical knee range of motion before aggressive strengthening can begin. Our current perioperative rehabilitation starts at the time of injury and preoperatively includes aggressive swelling reduction, hyperextension exercises, gait training, and mental preparation. Goals after surgery are to control swelling while regaining full knee range of motion. After quadriceps strengthening goals are reached, patients can shift to sport-specific exercises. When using a graft from the contralateral knee, the conflicting goals of strengthening the donor site and achieving full knee range of motion are divided between the knees. Thus, normal range of motion and strength can be achieved more easily and more quickly than when using an ipsilateral graft. Regardless of the graft source, a systematic rehabilitation program that emphasizes the return to symmetrical knee motion, including hyperextension, is necessary to achieve the optimum result. PMID- 16721539 TI - Current concepts of extracellular matrix. PMID- 16721541 TI - Invasive nontypeable Haemophilus influenzae infections in Germany: a case report of a previously healthy 7-year-old boy with an intracranial abscess, and epidemiological data from 2001 to 2004. PMID- 16721548 TI - An evaluation of thymidine phosphorylase as a means of preventing thymidine rescue from the thymidylate synthase inhibitor raltitrexed. AB - The antitumour effect of thymidylate synthase inhibitors such as raltitrexed (RTX) may be reversed by salvage of thymidine (Thd). Since thymidine phosphorylase (TP) depletes Thd, the potential for tumour-selective depletion of Thd using antibody-mediated delivery of TP to tumours was investigated. In vitro studies demonstrated that 25 x 10(-3) units/ml TP depleted extracellular Thd (3 microM) and restored sensitivity to the growth inhibitory effects of RTX in Lovo and HT29 cell lines. Thymidine concentrations in xenograft tumours were inversely proportional to the activity of TP in the tumour, and the presence of a subcutaneous Lovo xenograft reduced plasma Thd concentrations from 0.92 +/- 0.07 to 0.37 +/- 0.04 microM. Intravenous administration of native TP enzyme depleted plasma Thd to 5 nM, but following rapid elimination of TP, plasma Thd returned to pretreatment values. There was no effect on tumour TP or Thd. Conjugation of TP to the A5B7 F(ab)2 antibody fragment, which targets carcinoembryonic antigen (CEA) expressed on colorectal cell-lines such as Lovo, did result in selective accumulation of TP in the tumour. However, there was no tumour-selective depletion of Thd and there did not appear to be any potential benefit of combining antibody-targeted TP with RTX. PMID- 16721549 TI - Phase II study of docetaxel and cisplatin combination chemotherapy in metastatic gastric cancer. AB - PURPOSE: Docetaxel, as a single agent, has demonstrated activity in patients with advanced gastric cancer and cisplatin has shown lack of overlapping toxicities with docetaxel. Therefore, we conducted a phase II study to assess the efficacy and the toxicity of a combination regimen of docetaxel plus cisplatin in patients with advanced gastric cancer who have never been treated with palliative chemotherapy. METHODS: Ninety-two patients with metastatic gastric cancer were enrolled from April 2000 to March 2004. Patients with histologically confirmed gastric adenocarcinoma, at least one bi-dimensionally measurable lesion, no prior palliative chemotherapy and at least 6 months from the end of adjuvant chemotherapy were eligible for study entry. Docetaxel 75 mg/m(2 )and cisplatin 75 mg/m(2) were given on day 1. The cycle was repeated every 3 weeks. The objective response was evaluated after three cycles of chemotherapy. Toxicity was assessed according to the National Cancer Institute common toxicity criteria scale version 2.0. RESULTS: In total, 401 cycles were administered, with a median of 5 cycles per patient (range 1-9 cycles). The median age was 56 years (range 31-76). Eighty six patients were evaluable for treatment response. The objective response rate was 43.5% (95% CI, 33.4-53.6) with one complete response and 39 partial responses. Twenty patients (21.7%) had stable disease and 26 patients (28.3%) had a progression. The median time to progression was 7.0 months (95% CI, 5.0-9.0) and the median overall survival was 11.5 months (95% CI, 9.5-13.4). The chemotherapy was generally well tolerated and the most common grade 3-4 toxicities were neutropenia (17.4%), nausea/vomiting (13.0%) and diarrhea (7.6%). CONCLUSION: The combination chemotherapy of docetaxel with cisplatin in advanced gastric cancer was tolerable for most patients and showed a promising antitumor activity as a first-line therapy. PMID- 16721551 TI - Two-year evaluation of a new nano-ceramic restorative material. AB - The purpose of this prospective study was to evaluate the clinical performance of a new restorative material (Ceram.X) in combination with a new primer-adhesive (K 0127). One operator placed two Class I or II restorations in molars of 43 patients. One molar was restored with Ceram.X/K-0127, the other one with Tetric Ceram/Syntac Classic. At baseline, after 1 and 2 years, the restorations were evaluated by one evaluator using modified Ryge's criteria. After 2 years, 31 patients were examined. One Ceram.X-restoration had to be removed for root canal treatment due to pulpitis. Thus, failure rate of Ceram.X was 3.2% and of Tetric Ceram, 0%. In both groups, no sensitivity, no recurrent caries, and no changes in surface texture were recorded after 2 years. One restoration in each group showed slight changes in color stability (score B). Marginal discoloration (score B) was found concerning three Ceram.X-restorations (10.0%) and two Tetric Ceram restoration (6.5%). Marginal integrity was score B for four Ceram.X-restorations (13.3%) and for four Tetric Ceram-restorations (12.9%). No statistically significant differences were found (p>0.05). After 2 years of clinical service, 96.8% of Ceram.X/K-0127 and 100% of Tetric Ceram/Syntac Classic restorations were in place and performed clinically well. PMID- 16721552 TI - MRA of abdominal vessels: technical advances. AB - Magnetic resonance angiography (MRA) in general and MRA of the abdominal vessels in particular have undergone substantial improvements in the past 5 years triggered by the introduction and application of parallel imaging (PI), new sequence techniques such as centric k-space trajectories and undersampling, dedicated contrast agents and clinical high-field scanners. All of these techniques have the potential to improve image quality and resolution or decrease the image acquisition time. However, each of them has its own specific advantages and drawbacks. This review describes the main technical innovations and focuses on the impact these developments may have on abdominal MRA. Special consideration is given to the interaction of these various technical advances. The clinical value of advanced MRA techniques is discussed and illustrated by characteristic cases. PMID- 16721553 TI - Steal phenomenon through the anterior communicating artery in Moyamoya disease. AB - Branch occlusion of the anterior cerebral artery (ACA) is regarded as a part of Moyamoya disease. The purpose of this study is to define the ACA steal phenomenon (SP) in Moyamoya disease and to evaluate temoporal changes according to the disease progression. From 139 Moyamoya patients we defined ACASP as narrowing of the ipsilateral A1-2 junction while preserving the anterior communicating artery and supplying the contralateral ACA cortical branches with the development of leptomeningeal collaterals by the ipsilateral middle cerebral artery into the hypoperfused ipsilateral ACA territory. Direction of the steal related to the stage in both hemispheres by Suzuki classification was statistically analyzed using the binomial test based on binomial distribution. Follow-ups of ACASP were evaluated in five patients. We identified ACASP in 13 (9%) patients (male:female=7:6, mean age 18 years, range: 2-58 years) of the 139 study patients. The presenting pattern was ischemic in 12 and hemorrhagic in one. The direction of SP occurred from the hemisphere in the lower to the higher stage of Suzuki classification (two-tail P value=0.0002). After revascularization surgery, ACASP disappeared or diminished. ACASP may occur in bilaterally different stages of Moyamoya disease as a transient self-adaptive process. It regresses after revascularization surgery. PMID- 16721554 TI - Incidence of visualization of the normal appendix on different MRI sequences. AB - The purpose of this paper is to assess the incidence of visualization of the normal appendix on magnetic resonance imaging (MRI) examinations of the pelvis in a large adult population and to compare the yield of commonly used sequences. Pelvic MRI scans of 111 randomly selected patients were retrospectively reviewed by two fellowship-trained body imagers. Thirty-six cases, where the entire cecum and terminal ileum were not included in the field of view, were excluded. A normal appendix was definitively visualized in 55 of 71 patients on T1 spin echo (SE) sequences (78%). The appendix was seen on 25 of 42 (60%) half-Fourier single shot turbo spin echo T2. Visualization rates were 42% on pre-gadolinium T1 FS GRE, 54% on post-gadolinium T1 fat-suppressed gradient echo, and 17% of short tau inversion recovery sequences. MRI is an effective modality for visualization of the normal appendix. This may have important implications in patients with abdominal or pelvic pain, as visualization of a normal appendix should exclude the possibility of acute appendicitis from the differential diagnosis. PMID- 16721555 TI - TRPV1 (vanilloid receptor) in the urinary tract: expression, function and clinical applications. AB - The transient receptor potential vanilloid subfamily 1 (TRPV1) is an ion channel activated by capsaicin, heat, protons and endogenous ligands such as anandamide. It is largely expressed in the urinary tract of mammals. Structures in which the receptor expression is firmly established include sensory fibers and urothelial cells, although the presence of TRPV1 in other cell types has been reported. As in other systems, pain perception was the first role attributed to TRPV1 in the urinary tract. However, it is now increasingly clear that TRPV1 also regulates the frequency of bladder reflex contractions, either through direct excitation of sensory fibers or through urothelial-sensory fiber cross talk involving the release of neuromediators from the epithelial cells. In addition, the recent identification of the receptor in urothelial and prostatic cancer cells raise the exciting hypothesis that TRPV1 is involved in cell differentiation. Desensitization of the receptor by capsaicin and resiniferatoxin has been investigated for therapeutic purposes. For the moment, lower urinary tract dysfunctions in which some benefit was obtained include painful bladder syndrome and overactive bladder of neurogenic and non-neurogenic origin. However, desensitization may become obsolete when non-toxic, potent TRPV1 antagonists become available. PMID- 16721556 TI - P(2) purinoceptors account for the non-nitrergic NANC relaxation in the rat ileum. AB - The transmitters involved in the non-nitrergic component of the non-adrenergic, non-cholinergic (NANC) inhibitory response of the rat small intestinal longitudinal muscle to electrical field stimulation of its nerves is a matter of controversy. The present study is the first one to utilise a combination of a nitric oxide synthase inhibitor and a P(2) purinoceptor antagonist for studying this response. We found that the P(2) purinoceptor antagonist pyridoxalphosphate 6-azophenyl-2',4'-disulphonic acid (PPADS; 5x10(-5) M) abolished the non nitrergic NANC relaxation to electrical field stimulation (10 Hz). PPADS alone provided a significant, moderate inhibitory action. PPADS specifically inhibited relaxations due to exogenous adenosine 5'-triphosphate (ATP) or alpha,beta methylene ATP. The guanylate cyclase blocker 1H-[1,2,4]oxadiazolo[4,3 a]quinoxalin-1-one (ODQ; 10(-6) M) did not add to the inhibitory action of N(G) nitro-L-arginine on field stimulation-induced relaxation. ODQ abolished the relaxant effect of the nitric oxide donors nitroglycerin or sodium nitroprusside. These data indicate that: (1) nitric oxide and ATP fully account for the field stimulation-induced relaxation in the rat ileal strip under the experimental conditions of this study, and (2) no ODQ-sensitive guanylate cyclase-mediated mechanism is involved in the non-nitrergic component of the NANC relaxation. PMID- 16721557 TI - Treatment of intracranial stenoses using the Neuroform stent system: initial experience in five cases. AB - INTRODUCTION: We assessed the technical feasibility of balloon-assisted angioplasty with consecutive stenting using a flexible, self-expanding neurovascular stent for the treatment of intracranial arteriosclerotic vascular stenoses. METHODS: Five consecutive patients with symptomatic drug-resistant stenoses of the intracranial internal carotid artery (ICA) or the main stem of the middle cerebral artery (MCA) were treated by balloon-assisted angioplasty with consecutive stenting using the Neuroform stent system. RESULTS: Balloon dilatation of the stenoses and consecutive stent placement with complete coverage of the stenoses was feasible in all patients. One patient suffered acute thrombosis distally to the stented vessel segment which was successfully treated by fibrinolysis, and one patient suffered acute subarachnoid and parenchymal hemorrhage probably due to vessel perforation. In the other three patients, no complications occurred during or immediately after angioplasty. All patients were free of further ischemic events up to the 6-month follow-up. CONCLUSION: Our findings demonstrate that the Neuroform stent system can used successfully for the treatment of intracranial stenoses of the ICA and the main stem of the MCA. Although immediate angiographic results are promising, long-term angiographic and clinical follow-up is essential to demonstrate long-term outcome. PMID- 16721558 TI - CCR5 interactions with the variable 3 loop of gp120. AB - The G-protein coupled receptor CCR5 functions pathologically as the primary co receptor for macrophage tropic (R5) strains of HIV-1. The interactions responsible for co-receptor activity are unknown. Molecular-dynamics simulations of the extracellular and adjacent transmembrane domains of CCR5 were performed with explicit solvation utilizing a rhodopsin-based homology model. The functional unit of co-receptor binding was constructed via docking and molecular dynamics simulation of CCR5 and the variable 3 loop of gp120, which is a dominant determinant of co-receptor utilization. The variable 3 loop was demonstrated to interact primarily with the amino terminus and the second extracellular loop of CCR5, providing novel structural information regarding the co-receptor-binding site. Alanine mutants that alter chemokine binding and co-receptor activity were examined. Molecular-dynamics simulations with and without the variable 3 loop of gp120 were able to rationalize the activities of these mutants successfully, providing support for the proposed model. Based on these results, the global complex of CCR5, gp120 including the V3 loop and CD4, was investigated. The utilization of computational analysis, in combination with molecular biological data, provides a powerful approach for understanding the use of CCR5 as a co receptor by HIV-1. PMID- 16721559 TI - Molecular-dynamics simulations of pyronine 6G and rhodamine 6G dimers in aqueous solution. AB - We have carried out molecular-dynamics (MD) simulations on dimers of the positively charged laser dyes pyronine 6G (P6G) and rhodamine 6G (R6G) in aqueous solution, generating trajectories of 2.5 ns for various computational protocols. We discuss how the choice of atomic partial charges and the length of the trajectories affect the predicted structures of the dimers and compare our results to those of earlier MD-simulations, which were restricted to only 0.7 ns. Our results confirm that monomers of P6G easily undergo relative rotations within the dimer, but we found new conformations of the R6G dimer at longer simulation times. In addition, we analyzed in detail the energy change during the formation of dimers. With suitable corrections, the electrostatic energy from an Ewald treatment agrees with the results from an approach relying on a residue-based cutoff. For P6G, we show that the strong solvent-mediated electrostatic attraction between the monomers is counteracted by an almost equally large solvent-induced entropy contribution to yield a small driving force to dimer formation, in very good agreement with the free-energy change from a thermodynamic-integration procedure. Thus, earlier rationalizations of the dimer formation, based only on energy arguments, yield a qualitatively wrong picture. PMID- 16721562 TI - Sampling of water, soil and sediment to trace organic pollutants at a river-basin scale. AB - Sampling is considered a crucial step in the analysis of organic compounds in the environment. This review describes field sampling techniques and provides detailed step-by-step procedures for collection and preservation of all major environmental matrices (water, sediment and soil) integrated as part of the river basin water cycle. Attention is given to the prerequisites for obtaining reliable samples, and the practical issues of sample collection (planning, field sampling, sampling strategies and equipment and data quality assessment) are considered. Considering the heterogeneity of environmental matrices, special considerations for each matrix are given to solve typical problems and to find the most appropriate solutions to ensure the quality of the sample. The procedures described in the next sections are commonly used protocols that reflect true field conditions and current state-of-the-art techniques used in the sampling of organic compounds. The aim is to signify the importance of sampling to the overall analytical procedure. Finally, quality control issues to be considered in environmental sampling are given. PMID- 16721563 TI - [Phototherapeutic keratectomy. Undesirable effects, complications, and preventive strategies]. AB - The conceptual advantage of phototherapeutic keratectomy (PTK) include the option to remove diseased tissue on a micrometer scale. Although less invasive than other forms of corneal surgery, it remains a surgical procedure which carries inherent risks. In phototherapeutic indications this risk is higher than in otherwise healthy eyes undergoing photorefractive surgery. Potential general complications include wound-healing problems, infection, inflammation, or a recurrence of the underlying disease. More specifically, PTK is associated with unwanted refractive changes, such as a hyperopic shift or an irregular astigmatism. In this review, we discuss these problems along with prevention and treatment strategies in detail. PMID- 16721564 TI - [Unilateral hemiarcus lipoides corneae following filtration surgery]. PMID- 16721565 TI - [Analysis of ocular surface alterations following proton beam radiation in eyes with conjunctival malignant melanoma]. AB - BACKGROUND: In cases of large, diffuse or multilocular growth pattern of conjunctival melanoma, proton beam irradiation can serve as an alternative therapy to exenteration. In extended tumours, ocular surface problems can result after therapy. In this study we examined ocular surface integrity of ten patients who underwent proton beam radiation between 1996 and 2002. METHODS: The patients were examined during their follow-up. Eight of the ten cases who underwent proton radiotherapy were recurrent tumours, which were previously treated with other adjuvant therapies. We performed a standard ophthalmological examination and detailed tear film diagnostics. RESULTS: The follow-up was 17-87 months (mean: 40.9+/-20.1). In six cases more than 50% of the upper and lower eyelids were included in the radiation field. All of these cases showed moderate to severe sicca symptoms. The impression cytology revealed squamous metaplasia of conjunctival cells in nine of ten cases. CONCLUSIONS: Squamous metaplasia of conjunctival epithelia indicates a radiogenic, persisting disturbance of differentiation of the conjunctival epithelial cells. The tear film instability correlates with the loss of mucin-secreting goblet cells and meibomian gland dysfunction. PMID- 16721566 TI - Human radiation dosimetry of [11C]MeAIB, a new tracer for imaging of system A amino acid transport. AB - PURPOSE: [N-methyl-11C]alpha-methylaminoisobutyric acid ([11C]MeAIB) is a promising positron emission tomography (PET) tracer for imaging hormonally regulated system A amino acid transport. Uptake of [11C]MeAIB is totally specific for amino acid transport since [11C]MeAIB is metabolically stable both extra- and intracellularly. The aim of this study was to measure cumulated radioactivity in different organs and estimate the absorbed radiation doses to humans with the Medical Internal Radiation Dosimetry (MIRD) method. METHODS: Radiation absorbed doses were calculated from PET images for 25 volunteers. Dynamic acquisition data were obtained for the thoracic, abdominal, femoral and head and neck regions. The median dose of intravenously injected [11C]MeAIB was 422+/-35 MBq, with a range of 295-493 MBq. After PET imaging the radioactivity in voided urine was measured. Experimental human data were used for residence time estimates. Radiation doses were calculated with commonly used software. RESULTS: The effective dose for a 70 kg adult was 0.004 mSv/MBq, corresponding to a 1.72 mSv effective dose from the PET study with injection of 430 MBq [11C]MeAIB. The highest absorbed doses were in the pancreas (0.018 mGy/MBq), kidneys (0.017 mGy/MBq), intestine (0.014 mGy/MBq), liver (0.008 mGy/MBq) and stomach (0.005 mGy/MBq). Only 0.57% of injected activity was excreted to urine within 1 h after injection. CONCLUSION: Biodistribution of [11C]MeAIB in the abdominal region reflected the high activity of the transportation of amino acids via system A and these organs also had the highest radiation doses. An effective dose of 0.004 mSv/MBq is fully justified when [11C]MeAIB PET is performed to study system A activity in vivo. PMID- 16721567 TI - Is 3'-deoxy-3'-(18)F-fluorothymidine a better marker for tumour response than (18)F-fluorodeoxyglucose? AB - 3'-Deoxy-3'-(18)F-fluorothymidine (FLT) was developed in 1998 by Shields and co workers because monitoring of treatment response would be facilitated by imaging agents able to provide measures of tissue and tumour proliferation. Since then, FLT metabolism has been clarified in more detail in cell culture and experimental animal tumour models and also in clinical studies. Recently, FLT has increasingly been used for the assessment of response to anticancer treatment, mainly in tumour xenograft SCID mouse models; in contrast, clinical data are scarce. In this article we briefly summarise the intermediary metabolism of FLT and its application as an anticancer treatment response probe. The potential value and limitations of FLT as a highly promising proliferation imaging probe and its use for monitoring of treatment response are discussed. PMID- 16721568 TI - Imaging biomarkers as surrogate endpoints for drug development. AB - The employment of biomarkers (including imaging biomarkers, especially PET) in drug development has gained increasing attention during recent years. This has been partly stimulated by the hope that the integration of biomarkers into drug development programmes may be a means to increase the efficiency and effectiveness of the drug development process by early identification of promising drug candidates--thereby counteracting the rising costs of drug development. More importantly, however, the interest in biomarkers for drug development is the logical consequence of recent advances in biosciences and medicine which are leading to target-specific treatments in the framework of "personalized medicine". A considerable proportion of target-specific drugs will show effects in subgroups of patients only. Biomarkers are a means to identify potential responders, or patient subgroups at risk for specific side-effects. Biomarkers are used in early drug development in the context of translational medicine to gain information about the drug's potential in different patient groups and disease states. The information obtained at this stage is mainly important for designing subsequent clinical trials and to identify promising drug candidates. Biomarkers in later phases of clinical development may--if properly validated--serve as surrogate endpoints for clinical outcomes. Regulatory agencies in the EU and the USA have facilitated the use of biomarkers early in the development process. The validation of biomarkers as surrogate endpoints is part of FDA's "critical path initiative". PMID- 16721569 TI - The value of FDG-PET in patients with painful total knee arthroplasty. AB - PURPOSE: The purpose of this study was to evaluate 18F-fluorodeoxyglucose (FDG) uptake in patients with painful total knee arthroplasty and to relate FDG uptake to the location of soft tissue pain. METHODS: Twenty-eight patients with painful total knee arthroplasty had a clinical examination, standard radiographs, CT measurement of rotation of the femoral component and FDG-PET (18 PET/CT, 10 PET). The diagnosis of infection was based on microbiological examinations of surgical specimens (n=12) or clinical follow-up for at least 6 months (n=16), 99mTc labelled monoclonal antibody scintigraphy and joint aspiration. RESULTS: Twenty seven of 28 patients presented with diffuse synovial FDG uptake. Additional focal extrasynovial FDG uptake was observed in 19 knees. Twenty-four of the 28 patients had a diagnosis of internal femoral malrotation. The remaining four patients showed no rotation (0 degrees) and 3 degrees, 4 degrees and 7 degrees of external rotation, respectively. Three patients presented with the additional diagnosis of an infected total knee replacement. Pain was described as diffuse (n=10) or focal (n=18). In two knees a relationship between pain location and FDG uptake was observed. Of ten patients with a severe internal femoral component rotation (>6 degrees), seven had focal uptake, four in the femoral periosteum and three in the tibial periosteum. The difference between knees with severe malrotation and the remaining knees was not significant (p=1.000, Fisher's Exact Test). CONCLUSION: Diffuse synovial and focal extrasynovial FDG-PET uptake is commonly found in patients with malrotation of the femoral component and is not related to pain location. The information provided by FDG-PET does not contribute to the diagnosis and management of individual patients with persistent pain after total knee replacement. PMID- 16721570 TI - In vitro and in vivo targeting of different folate receptor-positive cancer cell lines with a novel 99mTc-radiofolate tracer. AB - PURPOSE: For the assessment of folate-based radiopharmaceuticals, human nasopharyngeal KB carcinoma cells are traditionally used although nasopharyngeal cancer is rare. On the other hand, the folate receptor (FR) is frequently overexpressed on diverse cancer types, the highest frequency (>90%) being on ovarian carcinomas. The goal of our study was the in vitro and in vivo assessment of different FR-positive human carcinoma cells. In addition, a murine sarcoma cell line was assessed as a pre-clinical alternative to human xenograft models. METHODS: FR-positive human nasopharyngeal, cervical, ovarian and colorectal cancer cell lines and the transgenic mouse sarcoma (24JK-FBP) cell line were targeted with a novel 99mTc-tricarbonyl folate derivative 2. Comparative in vitro cell binding studies were carried out under standardised folate-deficient conditions. In vivo studies were performed in nude mice and C6 black mice. RESULTS: The in vitro cell experiments revealed only FR-specific binding (unspecific <0.02%), ranging from 3.5% to 52% of complex 2 owing to variable levels of FR expression of the cell lines. In vivo tumour uptake of radiotracer 2 varied less than in vitro. It ranged from 0.66+/-0.17% ID/g (LoVo) through 1.16+/ 0.64% ID/g (IGROV-1) and 1.55+/-0.43% ID/g (24JK-FBP) to 2.33+/-0.36% ID/g (KB) 4 h p.i. CONCLUSION: These pre-clinical studies indicate that in vitro data obtained in FR-positive cancer cells do not necessarily correspond with or predict in vivo radiofolate uptake in corresponding (xeno)grafts. In addition, the murine 24JK-FBP cell line proved to be a valuable pre-clinical alternative to human tumour models. PMID- 16721571 TI - 99mTc-EDDA/HYNIC-octreotate scintigraphy, an efficient method for the detection and staging of carcinoid tumours: results of 3 years' experience. AB - PURPOSE: At all stages of the disease, serious difficulties are encountered in the imaging diagnosis of carcinoids. Somatostatin receptor scintigraphy (SRS) holds great promise for detecting primary tumours and metastases. 99mTc EDDA/HYNIC-octreotate should significantly improve the diagnosis of carcinoids in comparison with 111In-Octreoscan owing to the better affinity for SSR2 and the higher count rate. The aim of this study was to assess the diagnostic efficiency of 99mTc-EDDA/HYNIC-octreotate scintigraphy in the detection and staging of carcinoid tumours. METHODS: The study population comprised 75 patients (age 48.5+/-15.5 years): 46 with histological confirmation of carcinoid and 29 with suspected disease. 99mTc-EDDA/HYNIC-octreotate (740 MBq) SRS and CT were performed in all patients. Fifteen patients were examined with 111In-Octreoscan. RESULTS: High-quality 99mTc-EDDA/HYNIC-octreotate images were obtained in all cases, with maximum tumour tracer accumulation 4 h p.i. The mean target/non target ratios for whole body (WB) and SPECT scans were, respectively, as follows: primary lesions: 4.5 and 10.2; metastases: liver, 3.1 and 12.3; abdominal focal lesions, 2.7 and 5.8; lung, 2.7 and 8.3; mediastinum, 3.4 and 7.6; bones, 6.8 and 19.0. 99mTc-EDDA/HYNIC-octreotate WB scans revealed more metastases than 111In Octreoscan, with better individual separation. 99mTc-EDDA/HYNIC-octreotate SRS revealed new metastatic lesions in seven patients with confirmed carcinoid, and in four with dissemination the primary focus was found. Five patients qualified for radioguided surgery and 11 were referred to 90Y-DOTA-TATE therapy. The sensitivity of SRS in comparison with CT was higher for primary lesions and liver and abdominal lymph node metastases. In the subgroup of patients with suspected neuroendocrine tumours, two duodenal carcinoids, one thymic carcinoid and one ileal carcinoid were found. CONCLUSION: 99mTc-EDDA/HYNIC-octreotate, with high imaging quality, is an excellent alternative to 111In-Octreoscan for staging of carcinoids, and it seems to be the method of choice for detection of the primary focus in patients with metastases from an unknown primary tumour. PMID- 16721572 TI - Quantitation of cardiac sympathetic innervation in rabbits using 11C hydroxyephedrine PET: relation to 123I-MIBG uptake. AB - PURPOSE: Although (11)C-hydroxyephedrine ((11)C-HED) PET is used to map cardiac sympathetic innervation, no studies have shown the feasibility of quantitation of (11)C-HED PET in small- to medium-sized animals. Furthermore, its relation to (123)I-MIBG uptake, the most widely used sympathetic nervous tracer, is unknown. The aims of this study were to establish in vivo sympathetic nerve imaging in rabbits using (11)C-HED PET, and to compare the retention of (11)C-HED with that of (123)I-MIBG. METHODS: Twelve rabbits were assigned to three groups; control (n=4), chemical denervation by 6-hydroxydopamine (6-OHDA) (n=4) and reserpine treated to inhibit vesicular uptake (n=4). After simultaneous injection of (11)C HED and (123)I-MIBG, all animals underwent dynamic (11)C-HED PET for 40 min with arterial blood sampling. The (11)C-HED retention fraction and normalised (11)C HED activity measured by tissue sampling were compared with those measured by PET. RESULTS: Both the (11)C-HED retention fraction and the normalised (11)C-HED activity measured by PET correlated closely with those measured by tissue sampling (R=0.96027, p<0.001 and R=0.97282, p<0.001, respectively). Inhibition study by 6-OHDA resulted in a significant reduction in retention (90%) for both (11)C-HED and (123)I-MIBG. Reserpine pretreatment reduced (11)C-HED retention by 50%, but did not reduce (123)I-MIBG retention at 40 min after injection. CONCLUSION: Non-invasive quantitation of cardiac sympathetic innervation using (11)C-HED PET is feasible and gives reliable estimates of cardiac sympathetic innervation in rabbits. Additionally, although both (11)C-HED and (123)I-MIBG are specific for sympathetic neurons, (11)C-HED may be more specific for intravesicular uptake than (123)I-MIBG in some situations, such as that seen in reserpine pretreatment. PMID- 16721573 TI - Imaging of adrenocortical metastases with [11C]metomidate. PMID- 16721575 TI - Influence of the facial ruff on the sound-receiving characteristics of the barn owl's ears. AB - The barn owl, a nocturnal predator, derives its German name ("Schleiereule", direct English translation "veil owl") from the conspicuous ruff that covers the ear openings and gives the head a face-like appearance. The ruff is a specialization for the perception of sound. The densely-ramified reflector feathers forming the border of the ruff direct sound to the ear-openings. We studied the influence of the ruff on the behaviorally relevant sound-localization parameters interaural time difference (ITD) and interaural level difference (ILD). The directionality of the ear was much greater when the ruff was intact than when the reflector feathers were removed. With ruff intact, the distribution of ILDs was oblique and the maximum ITD occurred around 110 degrees of azimuth. When all head feathers were removed, the steepest ILD gradient was much closer to the horizontal axis and ITD was maximal at 90 degrees . Many effects were frequency specific. Thus, the ruff reflects some properties of the human pinna. However, by shifting the point where ITD becomes maximal beyond 90 degrees , the ruff also introduces a break of the front-back symmetry of ITD. PMID- 16721579 TI - Prenatal diagnosis of trisomy 18 as true mosaicism by three-dimensional ultrasonography: a case report. AB - Trisomy of chromosome 18 is the second most common autosomal trisomy, occurring in approximately 1:7,000 live births. Its prenatal diagnosis through abnormal findings in ultrasound with later analysis of fetal karyotype is important for a definition of the prognosis and counseling of the patients. We describe a case of trisomy 18 as true mosaicism diagnosed through amniocentesis in the second trimester of pregnancy, associated to the presence of multiple fetal phenotypic alterations. We focus on the importance of fetal morphological study through three-dimensional ultrasonography, which was highly important for clearly showing the fetus' structural alterations, helping parents to understand better the pathology and allowing them to reason about the continuity of the gestation. PMID- 16721580 TI - Biopsy-proven acute tubular necrosis in a child attributed to vancomycin intoxication. AB - Acute renal failure in children treated with vancomycin typically presents with interstitial nephritis. There is debate as to the extent of direct tubular toxicity attributable to vancomycin, especially in the absence of aminoglycoside treatment. We report a case of acute tubular necrosis (ATN) associated with vancomycin toxicity in an 8-year-old boy where there is no likely alternate explanation for toxic or ischemic injury. Treatment with hemodialysis resulted in the elimination of vancomycin from the circulation and subsequent improvement in renal function. PMID- 16721582 TI - Recurrence of proteinuria 10 years post-transplant in NPHS2-associated focal segmental glomerulosclerosis after conversion from cyclosporin A to sirolimus. AB - Mutations in the NPHS2 gene, which encodes podocin, are associated with steroid resistant nephrotic syndrome in childhood. Renal histology frequently presents focal segmental glomerulosclerosis (FSGS). Post-transplant recurrence of proteinuria in patients affected by homozygous or compound heterozygous NPHS2 mutation is encountered rarely (1-2%) compared to 30% recurrence in nonhereditary FSGS. We report on a pediatric kidney transplant recipient with NPHS2-associated nephrotic syndrome and FSGS, who developed biopsy-proven recurrence of FSGS 10 years post-transplant in temporal association with conversion from cyclosporin A (CsA)- to sirolimus (SRL)-based immunosuppression, due to histological evidence of severe CsA-induced nephrotoxicity. Reswitch of the immunosuppressive regimen from SRL to CsA led to a noticeable decrease of proteinuria and to stabilization of graft function. We conclude that patients with hereditary FSGS are not entirely protected from post-transplant recurrence of proteinuria, even in the long term. The close temporal relationship of FSGS recurrence with CsA withdrawal and conversion to SRL suggests that caution should be exercised in the use of CsA free immunosuppression also in patients with NPHS2-associated FSGS. PMID- 16721583 TI - Unexpectedly high exposure to enteric-coated mycophenolate sodium upon once-daily dosing. AB - Enteric-coated mycophenolate sodium (EC-MPS) has a mean half-life of 11.7 hours, which encouraged hope of using this drug once daily in a nonadherent adolescent SLE patient. This is a case report on a 17-year-old adolescent with a history of noncompliance who was switched from twice-daily mycophenolate mofetil (MMF) to once-daily EC-MPS. The EC-MPS dose was equimolar to the daily MMF dose (1 g MMF BID and 1.44 g of EC-MPS OD). The active compound of both drugs, mycophenolic acid, was measured using a commercially available EMIT assay. Both drugs were well-tolerated and maintained remission of the SLE. The average of three 12-hour areas under the time-concentration curves (AUC) on 1 g of MMF BID was 59.0 mgxh/L. In contrast, the 24-hour AUC after 1.44 g EC-MPS OD was 283.2 mgxh/L, more than double the expected 118.0 mgxh/L of two MMF dosing intervals. A repeat 24-hour AUC after 1.08 g of EC-MPS was 218.2 mgxh/L. EC-MPS once daily may be a well-tolerated therapeutic option for nonadherent adolescent lupus patients, but may be associated with a significantly higher exposure than the equivalent MMF BID dose. PMID- 16721584 TI - Urolithiasis, idiopathic hypercalciuria and insularity. PMID- 16721585 TI - Recurrence of Henoch-Schoenlein purpura nephritis after long-term remission in a 15-year-old girl. PMID- 16721586 TI - Serum cystatin C and left ventricular diastolic dysfunction in children with chronic kidney disease. AB - Previous studies indicate that serum cystatin C predicts incident heart failure in older adults. Children with chronic kidney disease (CKD) develop left ventricular (LV) diastolic dysfunction, often the initial abnormality of cardiac function. We hypothesized that cystatin C might predict LV diastolic dysfunction in children with CKD. Fifty-seven subjects, aged 6-21 years, with stage 2-4 CKD underwent echocardiography. Diastole was assessed from transmitral Doppler [maximum early (E wave) and late (A wave) diastolic flow velocities (E/A ratio)] and from tissue Doppler [septal mitral annular peak velocities (E')]. LV filling pressures were determined, using a ratio of E/E'. Fourteen (25%) patients had low E' and 15 (26%) had high E/E'. Children with abnormal E' or E/E' had significantly higher cystatin C levels than children with normal indices (P<0.05). Neither serum creatinine nor measured glomerular filtration rate (GFR) significantly correlated with E' or E/E'. Stepwise multiple regression analysis showed that cystatin C (beta=-0.825, P=0.023) and left ventricular mass (LVM) index (beta=0.099, P=0.006) independently predicted E'; LVM index independently predicted E/E' (beta=0.0173, P=0.01). We conclude that, in contrast to measured GFR or serum creatinine level, elevated serum cystatin C might be associated with diastolic dysfunction in children with CKD. PMID- 16721588 TI - Role of prostaglandins in the pathogenesis of X-linked hypophosphatemia. AB - X-linked hypophosphatemia is an X-linked dominant disorder resulting from a mutation in the PHEX gene. PHEX stands for phosphate-regulating gene with endopeptidase activity, which is located on the X chromosome. Patients with X linked hypophosphatemia have hypophosphatemia due to renal phosphate wasting and low or inappropriately normal levels of 1,25-dihydroxyvitamin D. The renal phosphate wasting is not intrinsic to the kidney but likely due to an increase in serum levels of fibroblast growth factor-23 (FGF-23), and perhaps other phosphate wasting peptides previously known as phosphatonins. Patients with X-linked hypophosphatemia have short stature, rickets, bone pain and dental abscesses. Current therapy is oral phosphate and vitamin D which effectively treats the rickets and bone pain but does not adequately improve short stature. In this review, we describe recent observations using Hyp mice; mice with the same mutation as patients with X-linked hypophosphatemia. We have recently found that Hyp mice have abnormal renal prostaglandin production, which may be an important factor in the pathogenesis of this disorder. Administration of FGF-23 in vivo results in phosphaturia and an increase in prostaglandin excretion, and FGF-23 increases proximal tubule prostaglandin production in vitro. In Hyp mice, indomethacin improves the phosphate transport defect in vitro and in vivo. Whether indomethacin has the same effect in patients with X-linked hypophosphatemia is unknown. PMID- 16721591 TI - Acute renal failure with encephalopathy following Salmonella enteritidis infection. PMID- 16721592 TI - Fabry disease and nephrogenic diabetes insipidus. AB - Fabry disease is a lysosomal storage disorder with kidney involvement. The initial manifestation of kidney disease is often impaired urinary concentrating ability in adolescence or young adulthood. We describe a boy diagnosed prenatally with Fabry disease who presented with polyuria, polydipsia, hypertension, hypokalaemia and proteinuria at 7 years of age. A formal water-deprivation test followed by vasopressin challenge confirmed nephrogenic diabetes insipidus. A renal biopsy revealed findings typical of Fabry disease. Angiotensin converting enzyme therapy resulted in rapid improvement of symptoms, normalization of blood pressure and resolution of hypokalaemia and proteinuria. This child is the youngest reported Fabry disease patient with documented renal pathology and clinical manifestations of hypertension, proteinuria and nephrogenic diabetes insipidus. PMID- 16721594 TI - Sodium modeling attenuates rises in whole-blood viscosity during chronic hemodialysis in children with large inter-dialytic weight gain. AB - Elevated whole-blood viscosity (WBV) is a risk factor for atherosclerosis and thrombosis. We analyzed WBV during hemodialysis (HD) in children and tested the hypothesis that sodium modeling (NaM) attenuates an increase in WBV. Each of six children underwent two control (C) and two NaM HD sessions, B and E. Rapid decline in sodium (Na) concentration occurred at the beginning of HD in B and at the end in E. We measured WBV at different shear rates (SRs) and documented the amount of fluid removed (FR), change in blood volume (BV), and hematocrit (Hct) before, during, and after HD. The percent increase of WBV in control sessions was significantly different at 2 h and 3 h during and after HD from baseline values. The mean percent change in WBV from baseline increased linearly over time during HD (R2>0.90). Hct, FR, and BV correlated with WBV (P<0.05). The effects of NaM on attenuation of WBV were statistically significant in three subjects with >5% inter-dialytic weight gain (IDWG) (P<0.05). WBV increased during HD in children. NaM appears to attenuate the rise in WBV in children with large IDWG. PMID- 16721595 TI - Loss of imprinting of the insulin-like growth factor 2 and the H19 gene in testicular seminomas detected by real-time PCR approach. AB - IGF2 and H19 are imprinted genes in normal human tissue, but many studies have observed a loss of imprinting (LOI) of these genes in tumors as an epigenetic alteration of the DNA, that leads to a biallelic expression predisposing cells to carcinogenesis and tumor growth. The aim of this study was to test the reliability of LightCycler-assisted Real-time PCR in detecting LOI of IGF2 and H19 in 39 patients with testicular germ cell tumors by comparing these results with the analysis generated by the golden standard restriction fragment length polymorphism (RFLP). With LightCycler-assisted Real-time PCR for IGF2 44% and for H19 49% of the patients were found to be heterozygous. This was consistent with the results obtained by RFLP, but surprisingly RFLP failed in more than 7% of the patients. In detecting LOI (for IGF2 in 41% and for H19 in 68% of the informative patients) the approach by RFLP was superior, since the results derived from LightCycler-assisted Real-time PCR showed reliable results in 76 and 10% of the samples concerning IGF2 and H19, respectively. Again, no discrepancy between the results obtained by the two methods occurred. In sum, LightCycler-assisted Real time PCR is a sufficiently working approach for the rapid and reliable detection of heterozygosity of IGF2 or H19 gene and identification of LOI of IGF2 and thus may be helpful in conducting large epidemiological studies. However, for the identification of LOI of the H19 gene in this cohort it possesses only restrictive use. PMID- 16721596 TI - The effects of low dose aluminum on hemorheological and hematological parameters in rats. AB - Aluminum (Al) is a nonessential element and humans are constantly exposed to Al as a result of an increase in industrialization and improving technology practices. Al toxicity can induce several clinical disorders such as neurotoxicity, gastrointestinal toxicity, hepatotoxicity, bone diseases, and anemia. This study aimed at evaluating the possible effects of short term and low dose Al exposure on hemorheological and hematological parameters in rats. Fourteen young, male Wistar albino rats were divided into two groups: 1 mg/200 g body weight of aluminum sulfate (Al(2)(SO(4))(3) was injected intraperitoneally to the first group for two weeks, three times a week. The animals of the control group received only physiological saline solution during this period. At the end of the experimental period, anticoagulated blood samples were collected and hematological parameters were determined using an electronic hematology analyzer. Red blood cell (RBC) deformability and aggregation were measured using an ektacytometer (LORCA) and plasma and whole blood viscosities were determined with a Wells-Brookfield cone-plate rotational viscometer. Significant decreases in mean corpuscular volume (MCV), red blood cell (RBC) deformability at low shear stress levels, the aggregation half time (t1/2) and the amplitude (AMP) of aggregation and significant increments in whole blood viscosity (WBV) at native and 40% hematocrit (Hct) of Al-treated rats have been observed. In conclusion, low dose Al(2)(SO(4))(3) exposure for a short-time may be responsible for alterations in either rheological properties of blood or hemorheological properties through a remarkable effect on RBC membrane mechanical properties . These alterations may also play an important role in the development of anemia in the Al-treated animals. PMID- 16721597 TI - Genetic evidence for 18S rRNA binding and an Rps19p assembly function of yeast nucleolar protein Nep1p. AB - The nucleolar protein Nep1 and its human homologue were previously shown to be involved in the maturation of 18S rRNA and to interfere directly or indirectly with a methylation reaction. Here, we report that the loss-of-function mutation Deltasnr57 and multicopy expression of the ribosomal 40S subunit protein 19 (Rps19p) can partially suppress the Saccharomyces cerevisiae Deltanep1 growth defect. SnR57 mediates 2'-O-ribose-methylation of G(1570) in the 18S rRNA. By performing a three-hybrid screen, we isolated several short RNA sequences with strong binding affinity to Nep1p. All isolated RNAs shared a six-nucleotide consensus motif C/UUCAAC. Furthermore, one of the isolated RNAs exactly corresponded to nucleotides 1553-1577 of the 18S rRNA, which includes G(1570), the site of snR57-dependent 18S rRNA methylation. From protein-protein crosslink data and the cryo-EM map of the S. cerevisiae small ribosomal subunit, we suggest that Rps19p is localized in close vicinity to the Nep1p 18S rRNA binding site. Our results suggest that Nep1p binds adjacent to helix 47 of the 18S rRNA and possibly supports the association of Rps19p to pre-ribosomal particles. PMID- 16721598 TI - FLO11 mediated filamentous growth of the yeast Saccharomyces cerevisiae depends on the expression of the ribosomal RPS26 genes. AB - The RPS26A and RPS26B isogenes of Saccharomyces cerevisiae encode two almost identical proteins of the small 40S ribosomal subunit, which differ by only two amino acid residues. Growth of an rps26BDelta mutant strain is normal, whereas an rps26ADelta strain displays a reduced growth rate and increased sensitivity towards the specific translational inhibitor paromomycin. An rps26ADelta rps26BDelta double mutant strain is inviable. RPS26A but not RPS26B is required for haploid adhesive and diploid pseudohyphal growth mediated by FLO11, which encodes an adhesion. The RPS26A and RPS26B transcripts make up about 70 and 30% of the cellular RPS26 mRNA, respectively. Overexpression of RPS26B, as well as an RPS26B open reading frame driven by the RPS26A promoter, complements the rps26ADelta deletion and restores haploid invasive growth as well as diploid pseudohyphal growth. These results suggest that the two proteins are functionally interchangeable. FLO11-lacZ activity is not present in haploid rps26ADelta yeast mutant strains, even though FLO11 mRNA levels are not reduced. This suggests that the amount of Rps26p is critical for accurate translation of the FLO11 mRNA, and therefore for the dimorphic switch of the bakera9s yeast from a single cell yeast to an adhesive filamentous growth form. PMID- 16721599 TI - Severe juvenile dermatomyositis: two patients complicated with extra musculocutaneous involvement. AB - Here we report two patients with severe juvenile dermatomyositis (JDM) complicated with extra musculocutaneous involvement. The first case (a 10-year old boy) had unusual initial presentation of JDM complicated with interstitial lung disease documented with high-resolution computed tomography. He had a rapidly progressive course and died in 7 weeks after the onset of the disease despite steroid and immunosuppressive treatment. The second case (a 14-year-old boy) was presented with myositis complicated with hepatitis. He also had a chronic course of JDM with unfavorable outcome. It appears that the prognosis of patients with severe JDM is related with the degree of autoimmune vasculitis on extra musculocutaneous involvement. PMID- 16721600 TI - Trypanosoma cruzi: experimental parasitism of bone and cartilage. AB - Trypanosoma cruzi causes Chagas' disease, a systemic infection that affects cells of meso-, endo-, and ectodermic origin. However, as far as we know, the presence of T. cruzi stages in bone has not been reported previously, and it has scarcely been investigated in cartilage. We inoculated 7- and 20-day-old (8 and 15 g) NMRI albino mice i.p. with metacyclic trypomastigotes from Rhodnius prolixus used for xenodiagnosis of mice previously infected with mammalian, human, and triatomines isolates, characterized by randomly amplified polymorphic DNA as zymodeme 1 (equivalent to T. cruzi I). Tissular parasitism (quantified according to the number of pseudocysts/50 fields 400x) showed amastigotes, intermediate forms, or trypomastigotes in sternum chondroblasts, osteoblasts, macrophages, and fibroblasts; chondrocyte and osteocyte invasion was rare. All isolates parasitized bone marrow macrophages, with few amastigotes. We observed marked associated myotropism, with or without inflammatory infiltration; there were small numbers of intensely parasitized mononuclear cells in perichondrium and periosteum. We discuss the results in relation to the marked differences of the T. cruzi tropism toward the different types of sternum cells, and, additionally, we outline the possibility of transmitting parasitized bone marrow through transplants. The fact of finding parasite stages in sternum bone and cartilage may be considered important due to the studies on Chagas' disease paleoparasitology that are based on histological and molecular analysis. PMID- 16721601 TI - The third newly discovered Eimeria species (Protozoa: Eimeriidae) described from wild reindeer, Rangifer tarandus, in Iceland. AB - Fecal samples from 56 adult reindeer from eastern Iceland were examined for coccidian parasites. One Eimeria species was found in an 8-year-old male. Prevalence of infection was 1.8%; oocyst per gram (opg) value was 150. The coccidium was identified and described as a new species. The sporulated oocysts are ellipsoidal and average size is 30.0 x 21.1 microm. The oocyst has two distinct walls. Wall thickness is approximately 1.0 microm, and the outer wall, approximately four-fifths of total thickness, is generally smooth and appears bicoloured. The outermost portion is light blue, and the innermost portion, yellow to pale brown. The inner wall is dark brown. Oocysts contain a prominent polar granule but are devoid of a micropyle. Oocysts enclose four spindle-shaped sporocysts with a rounded end opposite to the Stieda body. The average size of sporocysts is 15.3 x 6.5 microm. Sporocysts contain a granular sporocyst residuum that usually forms a cluster between the sporozoites and one large refractile body in each sporozoite. PMID- 16721602 TI - Molecular phylogenetic evidence of a haplosporidian parasite infecting the polychaete Syllis nipponica (Imajima, 1966). AB - A haplosporidian parasite was detected in the polychaete Syllis nipponica via the amplification and sequencing of the 18S rDNA locus. This provides additional evidence of haplosporidians in polychaetes, the host group from which the first species of the phylum were described about a century ago. In phylogenetic analyses, the parasite was determined to place among mollusk-specific taxa. As such, separate taxonomic groups for haplosporidians infecting different host phyla should not be anticipated. These findings also underscore the as-yet unknown life cycles of commercially important parasites. Attempts to visualize spores of the parasite directly with scanning electron microscopy did not succeed. PMID- 16721603 TI - The complete mitochondrial DNA sequence of the green alga Oltmannsiellopsis viridis: evolutionary trends of the mitochondrial genome in the Ulvophyceae. AB - The mitochondrial genome displays a highly plastic architecture in the green algal division comprising the classes Prasinophyceae, Trebouxiophyceae, Ulvophyceae, and Chlorophyceae (Chlorophyta). The compact mitochondrial DNAs (mtDNAs) of Nephroselmis (Prasinophyceae) and Prototheca (Trebouxiophyceae) encode about 60 genes and have been ascribed an 'ancestral' pattern of evolution, whereas those of chlorophycean green algae are much more reduced in gene content and size. Although the mtDNA of the early-diverging ulvophyte Pseudendoclonium contains 57 conserved genes, it differs from 'ancestral' chlorophyte mtDNAs by its unusually large size (96 kb) and long intergenic spacers. To gain insights into the evolutionary trends of mtDNA in the Ulvophyceae, we have determined the complete mtDNA sequence of Oltmannsiellopsis viridis, an ulvophyte belonging to a distinct, early-diverging lineage. This 56,761 bp genome harbours 54 conserved genes, numerous repeated sequences, and only three introns. From our comparative analyses with Pseudendoclonium mtDNA, we infer that the mitochondrial genome of the last common ancestor of the two ulvophytes closely resembled that of the trebouxiophyte Prototheca in terms of gene content and gene density. Our results also provide strong evidence for the intracellular, interorganellar transfer of a group I intron and for two distinct events of intercellular, horizontal DNA transfer. PMID- 16721604 TI - ITGB3 shows genetic and expression interaction with SLC6A4. AB - Autism affects more males than females and is associated with disturbances of the serotonin system. The integrin beta3 (ITGB3) and serotonin transporter (SLC6A4) genes were both recently identified as male quantitative trait loci (QTLs) for serotonin levels and alleles of each have been associated with autism. Here, we use publicly available genomic resources to determine whether regulation of expression level could be the mechanism behind association between serotonin level and noncoding variation in ITGB3. We also examine whether ITGB3 might interact with SLC6A4 to contribute to autism susceptibility. Using murine and human expression data, we observe that ITGB3 and SLC6A4 expression levels are correlated (0.38or=1 time per day had adjusted ORs of 0.74 (CI: 0.34-1.58), and 0.56 (CI: 0.32-0.98) in comparison with non daily green tea drinkers, respectively (p-trend <0.001). CONCLUSION: In a case-control study in Japan, we found that habitual green tea consumption may be strongly associated with a reduced risk for SAH. Our findings will be useful in targeting individuals and populations for the primary prevention of SAH. PMID- 16721634 TI - Interleukin 6 G-174 C polymorphism and breast cancer risk. AB - Interleukin-6 (IL-6) is a growth factor involved in many processes including carcinogenesis. The C allele of the G-174 C promoter single nucleotide polymorphism (SNP) in the IL-6 gene decreases levels of IL-6 expression and it has been studied in the context of breast cancer progression yielding contradicting results. Furthermore a recent study found that carriers of the C allele were at an increased risk for this disease. We aim to evaluate the association between this variant and breast cancer risk in Caucasian postmenopausal women. Women participating in the Rotterdam Study (N=3822), including 171 patients with breast cancer were genotyped for this polymorphism. In order to assess the relationship between this SNP and breast cancer we carried out a logistic regression in relation to the incidence of breast cancer. The C allele frequency was 41.3% and the genotypes followed Hardy-Weinberg distribution (p=0.3). The logistic regression analysis showed a slight increase of risk for C allele carriers (odds ratio=1.24, 95% CI: 0.8-1.9), compared to non-carriers of this allele. This increased risk was not statistically significant. Our data suggest that the IL-6 G-174 C polymorphism does not seem to play a role in breast cancer risk, although its role as a prognostic factor remains to be studied. PMID- 16721636 TI - Ventricular pseudoaneurysm after subacute myocardial infarction. AB - A 55-year-old woman with a blank cardiac history was admitted in a regional hospital because of acute left sided heart failure. Initial evaluation showed a subacute inferior wall myocardial infarction with preserved left ventricular function and moderately severe mitral regurgitation. Before referral, coronary angiography was performed. Ventriculography revealed a posterior pseudoaneurysm that was missed initially. Fortunately she survived two almost fatal episodes of cardiac asthma in that hospital. After the diagnosis was made, she was sent to our tertiary care hospital, where she was urgently and successfully operated. A short review, with emphasis on diagnosing left ventricular pseudoaneurysm is presented. PMID- 16721637 TI - Validation of a specific questionnaire on psychological and social repercussions of the lipodystrophy syndrome in HIV-infected patients. AB - OBJECTIVE: To validate a questionnaire to identify psychosocial repercussions of the lipodystrophy syndrome (LD) in HIV-infected patients. METHODS: A questionnaire containing 35 items in 6 subscales [body image, anxiety, depression, social support, social distress, quality of life (QoL)] was consecutively administered to 128 patients with LD, 25 naive patients or on therapy without LD, and to 15 HIV negative patients from January to July '04. Reliability was assessed using Cronbach's alpha and the intraclass correlation coefficient (ICC). The area under the curve (AUC) was used for discrimination between patients and control groups, and the scores were compared according to the severity of the LD. RESULTS: In all the subscales, the relationship between higher scores and LD severity was statistically significant (0.37-0.64, p<0.01), whereas the control groups had significantly lower scores. The internal consistency showed Cronbach's alpha coefficients ranging from 0.64 to 0.93. Test retest reliability after 15 days in 30 patients showed ICC values of 0.8-0.94. AUC values for the different subscales were higher than 0.7 with the exception of QoL score, and construct validity showed a significant correlation with the Beck Depression Inventory and the MOS-HIV scores (0.49 and 0.51, respectively, p<0.01). CONCLUSIONS: Our questionnaire shows satisfactory psychometric properties for identifying psychosocial repercussions of the LD in HIV-infected patients. PMID- 16721638 TI - Predictors of quality of life in HIV-infected rural women: psychometric test of the chronic illness quality of life ladder. AB - The Chronic Illness Quality of Life Ladder (CIQOLL) underwent psychometric testing in a sample of 278 women with HIV disease. The CIQOLL, a self-anchoring striving scale based on Cantril's Ladder, measures seven domains (physical , emotional, financial, family and friends, spiritual well-being, peace of mind, and overall life satisfaction) across four time periods (present, past, future, life without a diagnosis of HIV). The domains were derived from focus groups with persons with HIV disease. Women with a diagnosis of HIV Infection, age 18 or older, residing in rural areas in the southeastern United States, completed questionnaires that measured physical functioning, HIV related symptom frequency and distress, depressive symptoms, social support, and quality of life. Procedures used to assess reliability included item-item, item-total, and subscale-subscale correlations, and Chronbach's coefficient alpha. Criterion related (concurrent) validity was assessed by correlating the CIQOLL with HIV symptoms, functional status and social support. Construct validity was estimated using factor analysis and predictive modeling. Results provide preliminary evidence that the CIQOLL is a reliable and valid scale that may provide meaningful information about persons living with a chronic illness, such as HIV disease, especially low literacy and unacculturated populations. Additional research is needed to weight the domains, test the sensitivity of the scale to changes over time, and explore the usefulness of discrepancy scores. PMID- 16721639 TI - Scale for chemotherapy-induced long-term neurotoxicity (SCIN): psychometrics, validation, and findings in a large sample of testicular cancer survivors. AB - BACKGROUND: Neurotoxicity is common after modern chemotherapy, and can be assessed both objectively and subjectively based on symptoms reported by patients. The aim of this study was to explore the psychometric properties of a brief self-report scale for chemotherapy induced long-term neurotoxicity (the SCIN). METHODS: As part of a questionnaire survey 684 testicular cancer survivors (TCSs) filled in the SCIN, which assesses peripheral sensory neuropathy (paresthesias), Raynaud's phenomenon, and ototoxicity. Factor structure and internal consistency reliability of the SCIN were tested by the split-half method. In 538 TCSs, audiogram data were compared to the patients SCIN ratings of hearing. RESULTS: The internal consistency of the SCIN showed a Cronbach's alpha of 0.72. The three-factor structure of the SCIN was confirmed with 77% explained variance. All the SCIN items discriminated significantly between the TCSs who had been treated with cisplatin-based chemotherapy vs. those who did not receive such treatment. The individual cumulative cisplatin dose correlated significantly with all the SCIN items. The Pearson product-moment correlation coefficient was 0.54 between hearing reduction at 4000 Hertz measured by audiogram and the SCIN self report of reduced hearing. CONCLUSION: The SCIN shows good psychometric properties, and is recommended as a brief screening instrument for chemotheraphy indeed neurotoxicity. PMID- 16721640 TI - Repeatability of a Rasch model of the AQ20 over five assessments. AB - Rasch analysis is now used widely to assess the measurement properties of health status questionnaires. This study tested the stability of the AQ20 - a dichotomous-response measure of health status in asthma, using parameters estimated by a Rasch model. One hundred forty-four asthma patients completed the AQ20 on five occasions over 3 months. At visit 1, two items showed significant misfit and were deleted. At each visit, the overall mean item-person and item trait interaction statistics from the remaining 18 items (AQ18) were very similar. The repeatability of the item calibrations was excellent (intraclass correlation coefficient 0.95), despite the patients' health having changed (repeated-measures ANOVA: FEV(1) and AQ18 score p<0.0001). Tests of differential item functioning (DIF) over time showed that one item increased in severity. This item refers to ability to garden, and changes in response patterns could be related to seasonal changes over the study period. We conclude that this study has highlighted the usefulness of multiple repeat assessments which allow items to be tested for DIF over time. This is important as inclusion of 'time dependent' items in a questionnaire may reduce the reliability of the instrument. PMID- 16721641 TI - Health status in patients with Alzheimer's disease: an investigation of inter rater agreement. AB - OBJECTIVE: The purpose of the present study was to examine the level of agreement between health status ratings provided by patients with Alzheimer's disease and by their proxies. BACKGROUND: Because proxy-completed responses are often necessary in assessing health outcomes for the elderly, it is necessary to determine the feasibility and potential limitations of using proxies as a patient substitutes. METHODS: To assess the potential utility of proxy responses on health status when subjects present a cognitive impairment, this study compared the responses of 70 subjects with Alzheimer's disease and those of their family and/or care provider proxy using the SF-36. Agreement between proxies and patients was measured by intraclass correlation coefficients (ICCs). RESULTS: The proportion of exact agreement between patients and proxies on the 36 items ranged from 3.3 to 41.7%. Results reveal poor to moderate agreement between patient and proxy reports. Proxy reliability varied according to the relationship of the proxy to the index subject. Agreement decreased significantly with increasing severity of dementia and with increasing severity of Physical status (Katz ADL). Agreement was better for measures of functions that are directly observable and relatively poor for more subjective measures. CONCLUSIONS: Our results confirm the importance of the information source used for patient health status. PMID- 16721642 TI - Meaning of health-related quality of life among children and adolescents in an Asian country: a focus group approach. AB - AIMS: We aimed to evaluate the extent to which HRQoL instruments developed based on the Western notion of health is applicable to Asian children and adolescents by assessing their conceptualization of QoL. A secondary objective is to evaluate the necessity of developing age- or gender-specific HRQoL instruments. METHODS: We explored the meaning of general and health-related QoL through focus group discussions in Singapore, a multi-ethnic Asian society. Two investigators independently analysed the data. They extracted major and sub-themes related to both general and health-related QoL. The agreement between the analyses was compared and disagreement was resolved through discussion. RESULTS: Thirty-two subjects (children: 50.0%, female: 50.0%) participated. There were few disagreements. Meanings of general and health-related QoL could be grouped under three broad themes: (1) physical, (2) psychological and (3) social health, consistent with the current definition of HRQoL. We found that Singaporean and Western children/adolescents share a remarkably similar notion of general and health-related QoL. Compared to adolescents, children were more likely to report positive emotions, be less mindful of others' opinions and had less varied social activities. CONCLUSION: The results suggest that currently available instruments are potentially useful for Singaporean children/adolescents. We also found that age-specific HRQoL instruments are necessary. PMID- 16721643 TI - The Greek version of the Quality of Life in Epilepsy Inventory (QOLIE-31). AB - This study is presenting the translation and cultural adaptation into Greek of the Quality of Life in Epilepsy Inventory (QOLIE-31). We adapted the QOLIE-31 to Greek through a procedure of translation-back-translation. Sixty-three patients were interviewed and completed the QOLIE-31 and the GHQ questionnaires. We re examined a subset of them after a period of 2-5 weeks to evaluate the test-retest reliability of the questionnaire. We assessed the convergent validity by comparison of the QOLIE-31 and the GHQ and QOLIE-31 subscales and external measures. Discriminative validity was evaluated using the method of known-groups comparisons. The internal consistency was high for the QOLIE-31 and its' subscales (Cronbach's alpha 0.92 and 0.59-0.83 respectively). Test-retest reliability was acceptable (intra-class correlation coefficient 0.49-0.89 and Pearson's coefficient 0.53-0.92) for the group of patients who were re-examined. Comparison of the QOLIE-31 and GHQ scores showed agreement between the two questionnaires (Pearson's coefficient -0.61). We demonstrated the discriminative validity by the difference in the QOLIE-31 scores between patients with different seizure frequencies and different employment status. We concluded that the Greek version of the QOLIE-31 has psychometric properties equivalent to those of the original American-English version and is a valid and reliable instrument. PMID- 16721644 TI - Identification of domains relevant to health-related quality of life in patients undergoing major surgery. AB - This study examined themes relevant to health-related quality of life as identified by patients undergoing major surgery and by health care providers. Semi-structured interviews were conducted with 52 patients undergoing major abdominal, cardiac, orthopedic, thoracic, or vascular surgical procedures and 33 health professionals from various disciplines. A total of 58 themes were identified by content analysis. These themes were categorized into 6 domains. These were: physical well-being (14 themes), emotional well-being (13 themes), concern about quality of care (12 themes), social well-being (12 themes), cognitive preparation (4 themes), and spiritual well-being (3 themes). In general, we found that most aspects of health-related quality of life were common across various major surgical procedures and between patients and health care providers. However, when we examined the coverage of these themes in seven commonly-used health-related quality of life instruments, we found that many of the most frequently mentioned themes were not assessed with the available measures. A new evaluative instrument tailored to patients undergoing major surgery may therefore be warranted. PMID- 16721645 TI - How are quality of life ratings made? Toward a model of quality of life in people with dementia. AB - Quality of life (QOL) has become a focus of research in dementia. In measuring QOL, the views of people with dementia often have not been considered as researchers have proposed that they may not be able to articulate their opinions. This paper counters this belief, presenting a study using a grounded theory methodology to explore the issues that people with dementia felt were important for their QOL. Further, we propose a model of QOL including hypothesised links between important issues (including family and health), QOL and other variables. Twenty-five participants took part in one of nine focus groups. The groups included participants with mild to severe dementia with ages ranging from 49 to 93 years. Results indicate that most of the participants were willing and able to talk about their QOL. Of the 25 participants, only two said that their cognitive problems affected their QOL. Twenty-two issues were discovered through analysis to contribute to QOL, including continuingto live in your own home, independence, spouse and other family, feeling happy and feeling useful. People with dementia used social comparisons in talking about their QOL, as well as direct evaluation of their own happiness. A model of QOL based on theories of coping and response shift is suggested. The fact that so few of the participants talked about disease orientated issues challenges the large cognitive components included in many QOL measures for use with people with dementia. PMID- 16721646 TI - The significance of importance: an evaluation of Ferrans and Powers' Quality of Life Index. AB - Ferrans and Powers' Quality of Life Index (QLI) defines and assesses quality of life (QoL) in terms of importance-weighted life satisfaction. This study assessed the value of such weights and explored the relationship between weighted and unweighted (satisfaction only) scores and single-item rated overall life satisfaction (LS) and QoL. Data were collected by a postal survey to 81 Parkinson's disease patients (88% response rate). Correlations between weighted and unweighted QLI scores were >or=0.96, except for one subscale (r ( s ) = 0.85). Item non-response rates ranged between 4.2 and 45.1% and 1.4 and 38% for the weighted and unweighted QLI, respectively. Cronbach's alpha exceeded 0.7 for weighted and unweighted versions of two out of the four subscales and the total score. Scaling success rates were similar for weighted and unweighted scores and did not support the current subscale structure. Unexpectedly, weighted total scores correlated stronger with LS than with QoL, and unweighted scores displayed the opposite pattern. This study found no advantages by using importance-weighted satisfaction scores. The correlational pattern with overall LS and QoL challenges the QLI approach to QoL, although these observations may relate to the use of multiplicative item weights. This study has implications also beyond the QLI regarding, e.g., the use of multiplicative weights and the relationship between life satisfaction and QoL. PMID- 16721647 TI - Rhinitis Symptom Utility Index (RSUI) in Chinese subjects: a multiattribute patient-preference approach. AB - BACKGROUND: The Rhinitis Symptom Utility Index (RSUI), originally developed in the United States, consists of a patient-preference weighting scheme and a 10 item questionnaire measuring the severity and frequency of rhinitis related symptoms over a 14-day period. This study aimed to determine whether the Chinese RSUI could adopt the US-based multi-attribute utility function (MAUF) in scoring rhinitis symptoms. METHODS: In a Hong Kong study, 116 Chinese adults with allergic rhinitis completed the RSUI questionnaire and 36-item Short-Form Health Survey (SF-36) after they had been seen by two otorhinolaryngologists for disease severity ratings. Respondents then completed computer-administered direct preference measures, i.e., visual analogue scale (VAS) and standard gamble (SG) assessments. The VAS and SG data were used to estimate a MAUF for the Chinese based RSUI. RESULTS: The derived MAUF was somewhat different than the one developed for the US RSUI. Test-retest reliability for the Chinese RSUI was satisfactory (ICC = 0.71, p<0.001). Scores differentiated among cases with mild, moderate, and severe symptoms (p<0.001); and between those who did and did not require medications to control symptoms (p = 0.031). Findings were significantly correlated with SF-36 domain scores (r = 0.19 to 0.37; p=0.041 to <0.001). When the US-based scoring function was applied to the Chinese subjects, the resulting mean RSUI score was significantly lower (p<0.001). Comparisons between directly measured VAS and SG scores between the US and Chinese samples, demonstrated significant differences (all p<0.05), with the US subjects consistently rating rhinitis symptoms as worse than Chinese subjects. CONCLUSIONS: The Chinese RSUI has good measurement properties that reflect patient preferences from the Chinese. Results suggest that there are differences in preference rating between US and Chinese subjects and that use of the US-based preference function for the RSUI would bias the measurement of rhinitis symptom outcomes in Chinese subjects. PMID- 16721648 TI - A study of the construct validity of the Health Utilities Index Mark 3 (HUI3) in patients with schizophrenia. AB - This study assessed the construct validity of the Health Utilities Index Mark 3 (HUI3) in patients with schizophrenia. Patients with schizophrenia recruited from a tertiary mental hospital in Singapore completed the HUI3, the Short-Form 36 Health Survey (SF-36) and the Schizophrenia Quality of Life Scale (SQLS). Patients were assessed for presence and absence of 22 common psychiatric symptoms. Construct validity was assessed using 6 a priori hypotheses. Two hundred and two patients (mean age: 37.8 years, female: 52%) completed the survey. As hypothesized, overall HUI3 utility scores were correlated with SF-36 measures (Spearman's rho: 0.19 to 0.51), SQLS scales (Spearman's rho: -0.56 to 0.36), and the number of psychiatric symptoms (Spearman's rho: -0.49). The HUI3 emotion attribute was moderately correlated with SF-36 mental health (Spearman's rho: 0.45) and SQLS psychosocial scales (Spearman's rho: -0.43), and HUI3 pain attribute was strongly correlated with SF-36 bodily pain scale (Spearman's rho: 0.58). The mean HUI3 overall, emotion, cognition, and speech scores for patients with schizophrenia were 0.07, 0.09, 0.04 and 0.04 points lower than respective age-, sex- and ethnicity-adjusted population norms (p<0.001 for all, ANCOVA). This study provides evidence for the construct validity of the HUI3 in patients with schizophrenia. PMID- 16721649 TI - Validity and responsiveness of generic preference-based HRQOL instruments in chronic epilepsy. AB - Generic, preference-based instruments are recommended for assessing health related quality of life (HRQOL) in cost-utility analyses (CUA). We aimed to determine which instrument is the most appropriate for CUA of epilepsy care, using established psychometric criteria. We compared validity and responsiveness of EQ5D (using both UK and US preferences), visual analog scale (VAS), Health Utilities Index Mark II (HUI-2) and Mark III (HUI-3) and SF6D in 165 adults evaluated for epilepsy surgery. SF6D had the strongest or next-strongest associations with seizure severity and seizure control. It was not associated with education or IQ. Only SF6D and HUI-3 discriminated between patients with and without seizures 2 years after baseline evaluation. SF6D was most or next-most responsive to being seizure-free for 2 years, in most responsiveness analyses. VAS was also responsive, but showed less evidence of validity. The QOLIE-89, an epilepsy-targeted profile instrument, had stronger evidence for validity and responsiveness than the preference instruments. SF6D has several key psychometric advantages over four other preference instruments in CUAs of epilepsy care. This may reflect better coverage of HRQOL dimensions affected by epilepsy, greater sensitivity at the upper end of the HRQOL continuum, or both. These findings may not generalize to other chronic conditions. PMID- 16721650 TI - Factors associated with self-evaluated severity of illness and quality of life in male Israeli asthmatic soldiers. AB - OBJECTIVE: To evaluate whether the asthmatic condition and quality of life of male asthmatic Israeli soldiers deteriorate during army service. METHOD: We retrospectively assessed 178 asthmatic soldiers during service using an adapted version of the Mini Asthma Quality of Life Questionnaire, and compared it to their condition before service. The participants responded additionally to questionnaires assessing psychosocial resources and adjustment to illness. RESULTS: The self-evaluated degree of asthma disease severity (SEDOAD) of the participants deteriorated in the army compared to before service. Soldiers with mild SEDOAD during service, compared to soldiers with severe (and in most cases also moderate) SEDOAD, had less severe medically evaluated and self-evaluated asthma before service. Mild SEDOAD participants showed, additionally, lower inclination towards active seeking of asthma-related information, more favorable health care-related attitudes, less unfavorable effects of their illness on military and extra-military functioning, greater reported pre-enlistment optimism, and a greater belief in their ability to cope successfully with their asthma during service. CONCLUSION: SEDOAD and the quality of life during service are associated not only with pre-enlistment asthma severity, but also with the manner in which the soldier perceives the influence of the illness on his health condition, and the manner in which he copes with this influence. PMID- 16721651 TI - A survey of quality of life and depression for police officers in Kaohsiung, Taiwan. AB - OBJECTIVE: The enormous job stress of police work may result in depression, which is highly correlated with work disability and poor quality of life. We investigated the quality of life, the probability of depression, and the related risk factors for police officers in Kaohsiung, Taiwan. METHODS: We used the 12 item Short-Form Health Survey (SF-12) and the Disaster-Related Psychological Screening Test (DRPST) to assess the quality of life and prevalence of depression for 832 police officers in Kaohsiung. RESULTS: The estimated rate of probable major depression was 21.6% (180/832). Those with an educational level of university or above and nondepressed police officers had higher scores in every subscale for quality of life. Police officers older than 50 had higher scores in the mental aspects of quality of life. Family problems and job stress related to achievement, peer pressure about performance, and heavy workloads were predictive factors for depression. CONCLUSION: Police officers might have a higher estimated rate of depression than previously thought, and those with depression have a poorer quality of life. PMID- 16721652 TI - Measuring psychological consequences of screening: adaptation of the psychological consequences questionnaire into Dutch. AB - OBJECTIVE: To assess the psychometric properties of a Dutch adaptation of an originally Australian instrument measuring the psychological impact of breast cancer screening. METHODS: The three subscales (emotional, physical, social) of the Psychological Consequences Questionnaire (PCQ) underwent formal linguistic and cultural translation. A total of 524 women under intensive surveillance because of increased breast cancer risk were asked to complete the questionnaire at 2 months prior to screening, at the day of the screening visit preceding the screening, and 1-4 weeks after screening. Acceptability, score distribution, internal consistency, scale structure, responsiveness to change and construct validity were analysed. RESULTS: Response rates were high (98-94%) and there were very few missing answers and non-unique answers. All scales had Cronbach's alphas > 0.70. The physical and social subscale showed ceiling effects. The item-own scale correlations were only slightly higher than the corresponding item-other scale correlations. Factor analysis showed that the assumed three separate subscales were replicated in our study. Pre- and post-screening effect sizes for the emotional scale were larger than for the other two scales. All PCQ scales correlated with the scales of two other psychological measures (p 0) and occurred through the large increase in the binding entropy established when the hydrophobic bilirubin molecules escaped from their surrounding polar water molecules and into the hydrophobic medium of the first binding site. On the other hand, the calculated molar extinction coefficient illustrated that the microenvironment of the second binding set (especially for the third binding site) was less hydrophobic than the first one but still more hydrophobic than the buffer medium. The binding of the third bilirubin molecule to the HSA molecule was established more through exothermic (electrostatic) interactions. PMID- 16721656 TI - Structural features of transiently modified beta-lactoglobulin relevant to the stable binding of large hydrophobic molecules. AB - Binding sites for hydrophobic molecules on bovine beta-lactoglobulin, and their susceptibility to temperature, were studied by using various spectroscopic probes. Binding of probes carrying a single fluorophore moiety, a single nitroxide moiety, or both moieties on the same molecule, was followed by EPR and fluorescence. The presence of a fatty acid side chain in the dual probes was found to be required for binding to beta-lactoglobulin. Binding occurred only after the protein was heated at temperatures below the threshold for its irreversible denaturation. Binding became extremely tight and stable upon cooling of the protein-probe mixture. Comparison among the various probes suggests that multiple binding sites for hydrophobes are present in the native protein, and in the partially-and reversibly-modified form of beta-lactoglobulin present in solution at neutral pH and subdenaturing temperatures. Thus, the specificity of hydrophobes binding to beta-lactoglobulin may be modulated by simple physical treatment of the protein. PMID- 16721657 TI - Intersubunit disulfide interactions play a critical role in maintaining the thermostability of glucose-6-phosphate dehydrogenase from the hyperthermophilic bacterium Aquifex aeolicus. AB - Proteins from thermophilic microorganisms are stabilized by various mechanisms to preserve their native folded states at higher temperatures. A thermostable glucose-6-phosphate dehydrogenase (tG6PDH) from the hyperthermophilic bacterium Aquifex aeolicus was expressed as a recombinant protein in Escherichia coli. The A. aeolicus G6PDH is a homodimer exhibiting remarkable thermostability (t1/2 = 24 hr at 90 degrees C). Based on homology modeling and upon comparison of its structure with human G6PDH, it was predicted that cysteine 184 of one subunit could form a disulfide bond with cysteine 352 of the other subunit resulting in reinforced intersubunit interactions that hold the dimer together. Site-directed mutagenesis was performed on tG6PDH to convert C184 and C352 to serines. The tG6PDH double mutant exhibited a dramatic decrease in the half-life from 24 hr to 3 hr at 90 degrees C. The same decrease in half-life was also found when either C184 or C352 was mutated to serine. The result indicates that C184 and C352 may play a crucial role in strengthening the dimer interface through disulfide bond formation, thereby contributing to the thermal stability of the enzyme. PMID- 16721658 TI - Change of the protein p53 electrochemical signal according to its structural form - quick and sensitive distinguishing of native, denatured, and aggregated form of the "guardian of the genome". AB - Presence of mutated and/or structurally modified (e.g., denatured, aggregated) protein p53 form is associated with several disorders such as Alzheimer's disease, Parkinson's disease, prion diseases, and many types of tumours. The aim of this work was to distinguish native, denatured and aggregated form of full length p53 by flow injection analysis coupled with electrochemical detector (FIA ED). Firstly FIA-ED method used for protein native form determination was optimized (detection limit 45.8 amol per 5 mul injection; 3 x S/N). In addition the technique was applied to identify p53 structural forms (denatured and aggregated). It was found out that denatured form provides about three times higher electrochemical response (protein structure unfolding, approach of more electroactive centers - aminoacid residues - towards electrode surface) in comparison with native form. On the other hand, aggregated form offers lower response (steric eclipse of electroactive protein parts) when compared with the signal of native form. The obtained data show that we are not only able to sensitively determine native, denatured, and aggregated structural forms of p53 protein but also to distinguish them. PMID- 16721659 TI - Probing structure-function relationships of serine hydrolases and proteases with carbamate and thiocarbamate inhibitors. AB - Benzene-1,3-di-N-n-octylcarbamate (1), benzene-1-hydroxyl-3-N-n-octylcarbamate (2), benzene-1,3-di-N-n-ocztylthiocarbamate (3), and benzene-1-hydroxyl-3-N-n octylthiocarbamate (4) are synthesized from 1,3-benzene-diol and are characterized as the pseudo-substrate inhibitors of acetylcholinesterase, butyrylcholinesterase, cholesterol esterase, lipase, trypsin, and chymotrypsin. For these six enzyme inhibitions by 1-4, the pKi values are linearly correlated with their log ki values - Bronsted plots. Therefore, 1-4 inhibit these enzymes through a common mechanism. Moreover, both pKi and log ki values for the inhibitions by 1,3, and 4 are linearly correlated with both pKi and log ki values for the inhibitions by 2, respectively. Thus, the pKi values for the inhibitions by 2 are defined as the nucleophilicity constants of these enzymes (nenzyme). The log k2 values for the inhibitions by 1-4 are also linearly correlated with the nenzyme values. Therefore, the nucleophilicity for serine hydrolases and proteases toward 1-4 also applies the Swain-Scott correlations. PMID- 16721660 TI - Soret spectral and bioinformatic approaches provide evidence for a critical role of the alpha -subunit in assembly of tetrameric hemoglobin. AB - Soret spectral contributions of the alpha-subunit heme pocket have been evaluated by performing static titrations of apohemoglobin A with CNProtohemin under varied experimental conditions. Increasing the temperature from 5 to 30 degrees C in 0.05 M potassium phosphate buffer, pH 7, resulted in a decreasingly prominent hypsochromic shifts reflecting altered the vinyl-globin interactions. Studies at 10 degrees C in over pH range of 6.7-8.0 revealed a profile for the spectral shifts approximating the side chain pK value (7.4) a histidyl residue. These overall spectral changes correspond to DeltaE of < or = 7 kJ/mol indicative of electrostatic noncovalent interactions. Further our current molecular modeling studies indicate that the spatial arrangement and critical noncovalent interactions of tyrosine 42 and histidine 45 (aromatic residues unique to the alpha-subunit) make significant contribution to the Soret spectra. Most interestingly, phylogenetic analyses have revealed the presence of a histidyl triad in the alpha-chain of all vertebrates that form heterotetramers. PMID- 16721661 TI - Prediction of three dimensional structure of calmodulin. AB - Calmodulin (CaM) is an important human protein, which has multiple structures. Numerous researchers studied the CaM structures in the past, and about 50 different structures in complex with fragments derived from CaM-regulated proteins have been discovered. Discovery and analysis of existing and new CaM structures is difficult due to the inherent complexity, i.e. flexibility of 6 loops and a central linker that constitute part of the CaM structure. The extensive interest in CaM structure analysis and discovery calls for a comprehensive study, which based on the accumulated expertise would design a method for prediction and analysis of future and existing CaM structures. It is also important to find the mechanisms by which the protein adjusts its structure with respect to various factors. To this end, this paper analyzes the known CaM structures and finds four factors that influence CaM structure, which include existence of Ca2+ binding, different binding segments, measuring surroundings, and sequence mutation. The degree of influence of specific factors on different structural regions is also investigated. Based on the analysis of the relation between the four factors and the corresponding CaM structure a novel method for prediction of the CaM structure in complex with novel segments, given that the surroundings of the complex, is developed. The developed prediction method is tested on a set aside, newest CaM structure. The prediction results provide useful and accurate information about the structure verifying high quality of the proposed prediction method and performed structural analysis. PMID- 16721662 TI - Identification of eggshell membrane proteins and purification of ovotransferrin and beta-NAGase from hen egg white. AB - Exposure of selected Gram-positive and Gram-negative bacterial pathogens to egg shell membranes (ESM) significantly reduced their thermal resistance and/or inactivated cells. Although the components responsible for this antibacterial activity have not been conclusively identified, several proteins associated with the ESM activity have been identified including beta-N-acetylglucosaminidase, lysozyme and ovotransferrin, with each displaying varying degrees of antibacterial activity. Numerous attempts to purify active fractions of beta-N acetylglucosaminidase, lysozyme and ovotransferrin from the ESM proved somewhat limited; however, hen egg white (HEW) beta-N-acetylglucosaminidase was purified using a two-step chromatographic procedure, isoelectric focusing followed by cation exchange chromatography. Pure fractions of ovotransferrin were also obtained in the process. SDS-PAGE electrophoresis and Matrix-Assisted Laser Desorption Time-of-Flight Mass Spectrometry were then used to partially characterize the individual protein components. Purified protein fractions such as these will be required in order to fully elucidate the mechanism responsible for the antimicrobial properties associated with the ESM. PMID- 16721663 TI - Effects of tissue transglutaminase on beta -amyloid1-42-induced apoptosis. AB - Tissue transglutaminase (TGase) has been implicated in both cell survival and apoptosis. Here we investigate the role of TGase in beta-amyloid-induced neurotoxicity using retinoic acid (RA)-differentiated, neuronal SH-SY5Y cells. We show that beta-amyloid-induced cell death was reduced in RA-differentiated SH SY5Y cells treated with the TGase inhibitor monodansyl cadaverine. Expression of wild-type TGase enhanced beta-amyloid1-42-induced apoptosis, whereas transamidation-defective TGase did not. These effects were specific for beta amyloid-treated cells, as TGase reversed the neurotoxic effects caused by hydrogen peroxide treatment. Enhancement of beta-amyloid1-42-induced cell death by TGase was accompanied by marked increases in TGase activity in the membrane fractions and translocation of TGase to the cell surface. Overall, these findings suggest that the ability of TGase to exhibit pro-survival versus pro-apoptotic activity is linked to its cellular localization, with beta-amyloid-induced recruitment of TGase to the cell surface accentuating neuronal toxicity and apoptosis. PMID- 16721664 TI - Health care provider knowledge and practices regarding folic acid, United States, 2002-2003. AB - OBJECTIVE: To assess health care providers (HCP) knowledge and practices regarding folic acid (FA) use for neural tube defect (NTD) prevention. METHODS: Two identical surveys were conducted among 611 obstetricians/gynecologists (OB/GYNs) and family/general physicians (FAM/GENs) (2002), and 500 physician assistants (PAs), nurse practitioners (NPs), certified nurse midwives (CNMs), and registered nurses (2003) to ascertain knowledge and practices regarding FA. For analysis, T-tests, univariate and multivariate logistic regression modeling were used. RESULTS: Universally, providers knew that FA prevents birth defects. Over 88% knew when a woman should start taking folic acid for the prevention of NTDs; and over 85% knew FA supplementation beyond what is available in the diet is necessary. However, only half knew that 50% of all pregnancies in the United States are unplanned. Women heard information about multivitamins or FA most often during well woman visits in obstetrical/gynecology (ob/gyn) practice settings (65%), and about 50% of the time during well woman visits in family/general (fam/gen) practice settings and 50% of the time at gynecology visits (both settings). Among all providers, 42% did not know the correct FA dosage (400 mug daily). HCPs taking multivitamins were more than twice as likely to recommend multivitamins to their patients (Odds Ratio [OR] 2.27 95%, Confidence Interval [CI] 1.75-2.94). HCPs with lower income clients (OR 1.49, CI 1.22-1.81) and HCPs with practices having more than 10% minorities (OR 1.46, CI 1.11-1.92) were more likely to recommend supplements. NPs in ob/gyn settings were most likely and FAM/GENs were least likely to recommend supplements (OR 3.06, CL 1.36-6.90 and OR 0.64, CL 0.45-0.90 respectively). CONCLUSIONS: Knowledge about birth defects and the necessity of supplemental FA was high. Increasing knowledge about unintended pregnancy rates and correct dosages of FA is needed. The strongest predictor for recommending the use of FA supplements was whether the provider took a multivitamin. PMID- 16721665 TI - Validity of self-reported use of assisted reproductive technology treatment among women participating in the Pregnancy Risk Assessment Monitoring System in five states, 2000. AB - OBJECTIVES: To assess the validity of a question on assisted reproductive technology (ART) incorporated into the Pregnancy Risk Assessment Monitoring System (PRAMS) in 2000. While the intent of the question is to ascertain whether the index infant was conceived using ART, the phrasing was ambiguous for women who had used ART while trying to conceive the index infant but became pregnant after discontinuing treatment. METHODS: We compared weighted PRAMS estimates from five states that incorporated the ART question in 2000 with data from the U.S. ART Surveillance System (ART-SS) maintained by the Centers for Disease Control and Prevention (CDC). U.S. medical practices are mandated to report data for every ART procedure to CDC annually; thus, the ART-SS is highly specific and complete. RESULTS: ART use was reported for 156 of the PRAMS births in our study population, representing 4,571 (95% Confidence Limit, 3,452-5,690) births from the total birth cohort in the five states of interest in 2000. For the same maternal residency states and year, 1,768 births were reported to the ART-SS. Thus, we calculate that PRAMS overestimated ART use by 2,803 births. PRAMS estimated 2.59 times as many ART births as reported to the ART-SS. While for singletons, a large excess in estimated births from PRAMS was observed (ratio=3.50), there was little difference between the PRAMS estimates and ART-SS for twin and triplet births. CONCLUSION: These findings suggest women responding to PRAMS may be reporting past ART use in addition to current. The findings by plurality support this hypothesis. PMID- 16721666 TI - Selective migration and infant mortality among Puerto Ricans. AB - OBJECTIVE: This study examines the implications of migration to the United States for infant mortality among Puerto Rican mothers born in Puerto Rico. The roles of selective migration and duration of US residence are assessed. METHOD: Using survey data collected from mothers of infants sampled from computerized birth and infant death records of six US vital statistics reporting areas and Puerto Rico, we estimate logistic regression models of infant mortality among the sampled infants. These models provide a baseline for comparison with fixed-effects models based on all births within each mother's history. RESULTS: Logistic regression models for sampled infants show that the risk of infant mortality is lower for migrant women than for nonmigrant women in Puerto Rico until the migrants have lived in the United States for a substantial period of time. Fixed-effects models indicate that once unmeasured stable characteristics of the mother are controlled, early migrants do not differ from nonmigrants with respect to the risk of infant death. Both sets of models demonstrate that as mothers' exposure to the US mainland increases, the risk of infant mortality rises. CONCLUSIONS: Selective migration plays a role in the relatively low risk of infant mortality among recent Puerto Rican migrants to the United States. Migrants appear to be selected on qualities that contribute to favorable health outcomes for their offspring, but those qualities are later lost with exposure to life in the United States. PMID- 16721672 TI - Perspectives on tuberculosis among traditional Chinese medical practitioners in New York City's Chinatown. AB - This article discusses the perspectives of Chinatown's traditional Chinese medical practitioners on tuberculosis among New York City's Chinese laborers. The practice of traditional Chinese medicine (TCM) in the United States is neither regulated nor well understood. Some public health providers have expressed their concern that the use of TCM could prevent Chinese tuberculosis patients from receiving proper, biomedical treatment. Contrary to the suspicion of public health providers, the traditional Chinese medical practitioners in the context of New York City's Chinatown provide diverse methods of health care, many being familiar with the biomedical explanation for tuberculosis. All TCM informants in this study stated that biomedicine is more effective than Chinese medicine in treating tuberculosis. TCM in tuberculosis therapy is said to complement biomedicine and to restore bodily balance and the general health of patients. This study discusses the political-economic context shaping the explanation and treatment of tuberculosis among traditional Chinese medical practitioners and broadens our understanding of the various contexts in which TCM and biomedicine can be integrated. Furthermore, it is suggested that an opportunity exists for tuberculosis control programs to incorporate TCM practitioners in the effort to control the disease within New York City. PMID- 16721673 TI - The psychosocial function of reincarnation among Druze in Israel. AB - To gain an understanding of the psychosocial function of reincarnation among Druze, interviews were conducted with nine male subjects who had experienced reincarnation (Notq) and with one or two of their family members. Analysis of these interviews revealed that the onset of Notq typically occurs at between two and five years of age. Five of the subjects had displayed psychological distress in their childhood that was alleviated after the Notq. Once the child has displayed initial indications of reincarnation, such as mentioning names that the family construes as being from a past life, the family takes an active role in constructing the past-life story and matching it to a known real story involving a tragic death. This match creates a new order in the life of the child, the family, and the past-life family. All parties benefit from this new order: the child receives new special attention and love and becomes able to control and manipulate the parents; the parents are relieved because they see the child happy, and benefit from the social attention and regard they receive; and the grieving of the bereaved past-life family is alleviated by the realization that the soul of their lost son still lives. The findings support the sociocognitive notion of the constructing of past memories by the social environment. Additional research that should include data collection from the past-life family and examination of the affinity of reincarnation with dissociation and child abuse is recommended. PMID- 16721674 TI - Isla evangelista-a story of church and state: Puerto Rico's faith-based initiatives in drug treatment. AB - This article describes the debates leading to Puerto Rico's Mental Health Law of 2000, which defined addiction as a spiritual and social problem rather than a mental disorder, in order to trace three competing approaches to addiction in Puerto Rico: evangelist, biomedical, and harm-reductionist. Highlighting the ways in which the evangelist approach of Puerto Rican street ministries challenges the individualism underlying US faith-based initiatives and the punitive approach of the US War on Drugs, this article concludes that the virtues of the evangelist approach to addiction would be best supported by public funding for biomedical and harm-reduction approaches within a pluralistic system of treatment for addiction. PMID- 16721675 TI - Photosynthetic parameters as indicators of trinitrotoluene (TNT) inhibitory effect: change in chlorophyll a fluorescence induction upon exposure of lactuca sativa to TNT. AB - Trinitrotoluene, as a compound of conventional explosive, may cause inhibitory effect on terrestrial plants. When Lactuca sativa was exposed to different concentrations of trinitrotoluene (32-1000 mg/kg), photosynthetic process was investigated by using rapid chlorophyll fluorescence kinetic and pulse modulated fluorometry. The decrease of chlorophyll a variable fluorescence was seen to be caused by the deactivation of photosystem II reaction centers. We found for rapid variable fluorescence to be a useful indicator to evaluate the inhibitory effect of trinitrotoluene on photosystem II primary photochemistry and electron transport. The fluorescence parameters, related to the reduction state of photosystem II and to non-photochemical dissipation of light energy, showed a strong relation between the inhibitory effect of photosystem II activity and concentration of trinitrotoluene. The change of photosynthetic fluorescence parameters induced by trinitrotoluene was a reliable indication of the plant physiological state. We proposed for the reduction state of photosystem II and the non-photochemical energy dissipation to be a useful tool in bioassay toxicity testing of trinitrotoluene polluted soil. PMID- 16721676 TI - Autopathography and depression: describing the 'despair beyond despair'. AB - The Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, emphasizes diagnosis and statistically significant commonalities in mental disorders. As stated in the Introduction, "[i]t must be admitted that no definition adequately specifies precise boundaries for the concept of 'mental disorder' " (DSM-IV, 1994, xxi). Further, "[t]he clinician using DSM-IV should ... consider that individuals sharing a diagnosis are likely to be heterogeneous, even in regard to the defining features of the diagnosis, and that boundary cases will be difficult to diagnose in any but a probabilistic fashion" (DSM-IV, 1994, xxii). This article proposes that it may be helpful for clinicians to study narratives of illness which emphasize this heterogeneity over statistically significant symptoms. This paper examines the recorded experiences of unusually articulate sufferers of the disorder classified as Major Depression. Although sharing a diagnosis, Hemingway, Fitzgerald, and Styron demonstrated different understandings of their illness and its symptoms and experienced different resolutions, which may have had something to do with the differing meanings they made of it. I have proposed a word, autopathography, to describe a type of literature in which the author's illness is the primary lens through which the narrative is filtered. This word is an augmentation of an existing word, pathography, which The Oxford English Dictionary, Second Edition, defines as "a) [t]he, or a, description of a disease," and "b) [t]he, or a, study of the life and character of an individual or community as influenced by a disease." The second definition is the one that I find relevant and which I feel may be helpful to clinicians in broadening their understanding of the patient's experience. PMID- 16721677 TI - [Recurrence patterns and surgical treatment for recurrent colon cancer after radical resection]. AB - OBJECTIVE: To investigate the surgical treatment for recurrent colon cancer after radical resection. METHODS: Clinical data of 87 cases with recurrence colon cancer after radical resection from Jan. 1999 to Dec. 2005 were analyzed retrospectively. RESULTS: The resection rate of recurrent colon cancer was 74.7% . 55.2% (48/87) of the cases received radical resection,and the median survival was 49 months,while 19.5% (17/87) received palliative resection with a median survival of 24 months, 25.3% (22/87) only exploration or by- pass operation with a median survival of 10 months. There were significant differences in survival among the different surgical treatments (P=0.003). CONCLUSION: The resection rate of recurrent colon cancer is high,and reoperation can prolong the survival of such patients. PMID- 16721678 TI - [Effect of total mesorectal excision and preoperative chemoradiotherapy on local recurrence in rectal cancer]. AB - OBJECTIVE: To investigate the effect of total mesorectal excision(TME) and preoperative therapy on local recurrence in rectal cancer. METHODS: Rectal cancer patients who received TME in School of Oncology, Peking University, from January 2000 to August 2004 were enrolled in the study group. Patients who received surgical resection for rectal cancer from January 1996 to December 1999,before the introduction of TME,were chosen as controls. Postoperative complications and local recurrence were compared. Clinicopathological and follow- up data were analyzed. RESULTS: There were 161 patients in the TME group and 173 as controls. The intra- operative blood loss was significant less,hospital stay shorter,and lymph nodes harvested more in TME group than those in the control group,there was no difference in complication rate between the two groups. Local recurrence (LR) rate was 2.5% in TME and 8.0% in the control group, respectively (chi2=5.144; P=0.023). In TME group,the local recurrence rate was 1.8% in the 77 patients with preoperative therapy,and 2.9% in the other patients without preoperative therapy (P=0.182). Logistic regression analysis revealed that TME and vessel cancerous emboli were major risk factors for local recurrence of rectal cancer. CONCLUSION: TME and vessel cancerous emboli are major risk factors for local recurrence of rectal cancer. PMID- 16721679 TI - [Risk factors for recurrence and metastasis after radical anterior resection for rectal cancer]. AB - OBJECTIVE: To investigate the risk factors for local recurrence and distant metastasis after radical anterior resection for rectal cancer. METHODS: Clinicopathological data of 957 patients who underwent radical anterior resection for rectal cancer from 1983 to 2000 were reviewed retrospectively. The risk factors for local recurrence and distant metastasis were analyzed. RESULTS: There were 150 recurrent or metastatic cases (15.7%) after radical resection during a median follow- up of 71 months. Recurrence and metastasis sites included pelvics(6.0%, n=57), liver (4.9%, n=47), lung (4.2%, n=40) and other sites (0.6%, n=6). The median recurrent interval was 18 months (2-85 months),with a median survival of 8 months (1-62 months) after recurrence. Re-resection of the tumors was performed in 23 patients(15.3% ), and the median survival of such patients was 30 months with a 5- year survival rate of 13.0%. There were significant differences in recurrence and metastasis considering age,family history of tumor,CEA level,T staging,lymph node metastasis,venous cancerous emboli and signet cell carcinoma or mucinous adenocarcinoma. Logistic regression analysis revealed that family history (P=0.001), high CEA level (P=0.033), T3- 4 (P=0.000), lymph node metastasis (P=0.000),venous cancerous emboli (P=0.001),and signet cell carcinoma or mucinous adenocarcinoma (P=0.012) were risk factors for recurrence and metastasis. CONCLUSIONS: There are several risk factors for recurrence after radical anterior resection for rectal cancer. The main recurrent or metastatic sites are pelvis,liver and lung. Resection of recurrent tumor can prolong the survival. PMID- 16721680 TI - [Indications for colonoscopy examination and its disease distribution: a report of 5690 cases]. AB - OBJECTIVE: To explore the indications for colonoscopy examination and the distribution of diagnostic diseases. METHOD: From Jan. 2000 to Dec. 2004, 5960 patients received colonoscopy examination in our colorectal center. The indications for colonoscopy examination and the distribution of its diagnostic diseases were analyzed. RESULTS: There were 3096 males and 2594 females,and the mean age was (52+/-15) years. The reasons for colonoscopy included hemafecia (26.9%), atypical abdominal pain (25.8%), diarrhea or increased frequency of stool (11.1%), anal tenesmus or discomfort (7.6%), constipation (7.0%),mucous or bloody purulent stool (3.0%), intra-rectal mass or abdominal mass on physical examination (0.9%), re- examination after colonoscopic polypectomy (10.9%), re examination after operation for colorectal cancer(1.5%), simple health examination (2.2%). Colonoscope reached the cecum in 97.7% of the cases,and at least one disease was found in 2283 cases (40.1%). Among them,colorectal cancer accounted for 10.3%, colorectal polyps 19.6%, ulcerative colitis 4.3%, and Crohn's disease 0.5% respectively. CONCLUSION: The indications for colonoscopy are too strict to screen the early stage colorectal cancer. Colonoscopy should be performed in the cases with symptoms such as bloody stool, diarrhea, abdominal pain, constipation, or with colorectal polyps, after operation for colorectal cancer,or as members of hereditary colorectal cancer family. PMID- 16721682 TI - [Comparison of surgical complication rate between laparoscopic and open radical resection for colorectal cancer]. AB - OBJECTIVE: To compare the surgical complication rate between laparoscopic and open radical resection for colorectal cancer. METHODS: From September 2000 to December 2005, 491 cases with colorectal cancer were divided into two groups prospectively and nonrandomly,and received radical laparoscopic operation (LP, n=214) and open operation (OP, n=277). The intra- and post-operative complication rate were compared between the two groups. RESULTS: In laparoscopic groups, 14 cases (6.54%,14/214) were converted to open surgery,because of intra-operative complications in 7 cases,obesity or large tumors in 5 cases,narrow- pelvis in one case and retroperitoneal tumor in one case. The intra-operative complication rate was 4.8% (10/207) in LP group and 3.6% (10/277) in OP group (P > 0.05, chi2=0.446). There were no differences in post- operative intestinal obstruction, stoma leak, stoma bleeding, chyle leak, pulmonary infection except incision infection(5.5% vs 14.1%, P< 0.05) between LP and OP groups. The overall postoperative complication rate was 23.5% (47/200) in LP group and 36.8% (102/277) in OP group (P< 0.01, chi2=9.598). CONCLUSIONS: There is no difference in intra-operative complication rate between LP and OP groups,but the post operative complication rate is significantly lower in LP group than that in OP group. PMID- 16721681 TI - [Value of lymph node micrometastasis detected by RT-PCR assay in determining the stage of gastric carcinoma]. AB - OBJECTIVE: To detect micrometastasis in regional lymph nodes using RT- PCR assay and evaluate the significance of the new assessment of nodal status in determining pN staging in gastric carcinoma. METHODS: In addition to HE,RT- PCR assay for cytokeratin-20 gene marker was used to detect micrometastasis in 850 lymph nodes from 30 patients with gastric carcinoma who underwent gastrectomy with lymphadenectomy from Dec. 2003 to Apr. 2004. RESULTS: The detection rate of HE staining was 27.1% (233/850), the detection rate of RT-PCR assay was 36.5% (310/850) (Pa< 0.01). Lymph node micrometastasis was further detected in 77 nodes from 14 patients. The detection rate of RT- PCR assay from the routine negative lymph nodes was 12.5% (77/617). Seven of those patients were up- staged (from IB stage to II stage, from IB stage to III A stage, from I stage to III A stage, from III A stage to III B stage, from III A stage to IV stage in one patient respectively, and from III B stage to IV stage in 2 patients). CONCLUSION: RT- PCR assay can increase the detection rate of lymph node metastasis and have a significant impact on the staging system of gastric carcinoma. PMID- 16721683 TI - [Meta analysis of 2025 cases with multiple primary colorectal carcinoma]. AB - OBJECTIVE: To summarize the clinicopathological characteristics of multiple primary colorectal carcinoma. METHOD: With "multiple primary colorectal carcinoma" as search words,and with the same inclusion and exclusion criteria, related Chinese literatures published were retrieved. A meta analysis was carried out. RESULTS: The incidence of multiple primary colorectal carcinoma was 2.9%; and the average age was 53 years. 55.2% of the cases with synchronous multiple primary colorectal carcinoma were diagnosed by fiberoptic colonoscopy before operation,37.5% by intraoperative exploration, 15.7% by postoperative pathology. 94.8% of the cases with metachronous multiple primary colorectal carcinoma were diagnosed by fiberoptic colonoscopy before operation,and the average interval was 5.2 years. 30.9% of the lesions were located in the rectum, 19.9% in the sigmoid, 9.0% in the descending colon, 5.2% the in splenic flexure, 9.1% in the transverse colon, 6.1% in the hepatic flexure, 11.8% in the ascending colon,and 8.1% in the caecum. 37.6% of the cases were complicated with extra- intestinal lesions,and 43.7% adenoma or polyps. 34.6% of the cases had lymph node metastasis. Histological type was the same in 60.6% of the cases,and adenocarcinoma accounted for 89.2% and cancerization of adenoma 8.4%. Radical resection was performed in 85.9% and palliative resection in 10.1% of the cases. The 3, 5, 10 and 15 year survival rates were 64.3%, 44.6%, 26.3% or 9.4% in synchronous multiple primary colorectal carcinoma respectively, and 69.6%, 59.2%, 45.0%, 36.7% in metachronous multiple primary colorectal carcinoma. CONCLUSIONS: Multiple primary colorectal carcinoma is rare in clinic. The diagnosis mainly depends on colonoscopy and intra-operative exploration. The resection rate is high and the prognosis is better. The prognosis of metachronous multiple primary carcinoma is better than that of synchronous multiple primary colorectal carcinoma. PMID- 16721684 TI - [Clinicopathologic features of the patients with node-negative metastasis gastric adenocarcinoma and their survival analysis]. AB - OBJECTIVE: To investigate the clinicopathologic features of the patients with node-negative metastasis (pN0) gastric carcinoma confirmed by routine pathologic examination (Ha&E staining),and their relationship with survival. METHODS: The clinico-pathologic data of 87 pN0 gastric carcinoma patients were analyzed retrospectively. Kaplan-Meier (Log-rank) method was used to compare the survival rate,and Cox regression method was used to screen the independent prognosis factors for pN0 gastric cancer. RESULTS: There were 69 males and 18 females. D(2) lymphadenectomy was performed in 75 (86.2%), and D(3) in 13 cases (13.8%). Distal gastrectomy was performed in 59 (67.8%), total gastrectomy in 24 (27.6%) and proximal gastrectomy in l4 cases (4.6%). Twenty-three (26.4%) cases received postoperative chemotherapy and 64 (73.6% ) did not. The 5-year survival rate of the 87 pN0 gastric carcinoma patients was 56.5%. There were no differences in survival rate considering age, tumor location, gastrectomy, lymph nodes dissection, differentiation grade, and tumor-stroma relationship (P > 0.05), while the differences were significant in survival rate between the patients with postoperative chemotherapy,lymphatic invasion,venous invasion or not,and different pT staging (all Pa< 0.05). The Cox regression analysis revealed that pT staging and postoperative chemotherapy were independent prognostic factors. CONCLUSION: pT staging and postoperative chemotherapy are independent prognostic factors. It is necessary to develop a precise staging technique to select a suitable surgical therapy or confirm which patients should accept postoperative chemotherapy in pN0 gastric carcinoma patients. PMID- 16721685 TI - [Hyperthermia enhanced the killing effect of 5-fluorocytosine on human colon cancer cell line transfected with cytosine deaminase gene]. AB - OBJECTIVE: To investigate whether hyperthermia can enhance the killing effect of 5- fluorocytosine (5- FC) on human colorectal carcinoma cell lines SW480 transfected with carcinoembryonic antigen (CEA) tissue- specific cytosine deaminase (CD) gene in vitro,and study its mechanism. METHODS: Human colorectal carcinoma cell lines SW480 transfected with G1CEACDNa were cultured. The proliferated colonies were treated with the combined therapy of 5-FC and hyperthermia at a temperature of 43 degrees C for 30 min. After eight days, MTT was used to calculate the cellular survival rate,to analyze the killing effect of 5-FC combined with hyperthermia on SW480 cells transfected with CD gene. Flow cytometry was performed to analyze the cellular cycle and transmission electron microscope was used to observe the morphologic changes of SW480 cells after thermochemotherapy. RESULTS: Hyperthermia combined with 5-FC had an enhanced killing effect on SW480-CEACD cells than 5-FC alone (P< 0.05, t =2.403, n=9). Flow cytometry revealed that the proportion of S stage cell increased in the group treated with hyperthermia and 5- FC (P< 0.001, t =7.158, n=6). Transmission electron microscope showed apoptosis after thermo- chemotherapy. CONCLUSIONS: Hyperthermia can improve the anti- tumor effect of 5- FC on human colorectal carcinoma cell lines SW480 transfected with CD gene, and the cells were blocked at S stage of cellular cycle and apoptosis was induced following thermochemotherapy. PMID- 16721686 TI - [Clinical study of continual jejunal interposition after subtotal gastrectomy]. AB - OBJECTIVE: To explore the clinical effect of continual jejunal interposition in digestive tract reconstruction after subtotal gastrectomy. METHODS: Thirty-four patients with distal gastric cancer were divided randomly into two groups. In group A, the digestive continuity was reconstructed by continual jejunal interposition in 16 patients after subtotal gastrectomy. In group B, the digestive tract of other 18 cases were reconstructed by Billroth II procedure. The postoperative comp1ications, nutritional status, food intake and gastroscopic results were compared. RESULTS: There were no complications such as anastomotic leakage or obstruction in the two groups. The Visick scoring of group A was better than that of group B, and the difference was significant one year after operation (mu= 1.98, P< 0.05). All patients retrieved 85% of preoperative food intake per meal in group A, while only l4 patients got such results in group B. The weight loss was significantly higher in group B than that in group A(t = - 2.181, P= 0.037) after operation. The serum albumin level after operation in group A was significantly higher than that in group B (t=2.125, P=0.041), the level one year after operation in group A was also significantly higher than that before operation (t= - 2.175, P= 0.011). Gastroscopy one year after operation revealed fluent stoma,no bile reflux,and no congestion and edema in stomal mucosa and interposed jejunum in group A, while bile retention in 11 cases (61.1%), stomal inflammation in 13 cases (72.2%), and stomal ulcer in 2 cases (11.1%) in group B. CONCLUSION: Continual jejunal interposition after subtotal gastrectomy can recover physiological continuity of digestive tract and improve the quality of 1ife without reflux gastritis. PMID- 16721687 TI - [Comparison of efficacy between procedure for prolapse and hemorrhoids and open hemorrhoidectomy]. AB - OBJECTIVE: To compare the results of procedure for prolapse and hemorrhoids (PPH) and open hemorrhoidectomy. METHODS: A standard questionnaire was given to all patients after PPH or open hemorrhoidectomy from March 2001 to March 2004. In combination with proctological examination, the results including symptoms relief and recurrence were compared between the two groups. RESULTS: There were 184 effective questionnaires, including 96 cases in PPH group and 88 in open hemorrhoidectomy group. PPH and open hemorrhoidectomy both relieved prolapse (92.7% vs 96.8%, P=0.282), bleeding (91% vs 81%, P=0.241) and pain (91.7% vs 91.5%, P=0.977). There were no statistical differences in the overall complication rate (30.2% and 29.5%, P=0.923) and recurrence rate (21.8% vs 20.5%, P=0.814) between the two groups. The overall satisfactory degree was 87.5% in PPH group and 84.8% in open hemorrhoidectomy group (P=0.218). CONCLUSION: PPH is a safe and effective option for prolapsed hemorrhoids compared with open hemorrhoidectomy. PMID- 16721688 TI - [Effect of the width of excisional mucosa stoma ring on postoperative results of procedure for prolapse and hemorrhoids method]. AB - OBJECTIVE: To explore the effect of the width of excisional mucosal stoma ring on postoperative results of procedure for prolapse and hemorrhoids (PPH) in the treatment of hemorrhoids. METHODS: Four hundred and eighty hemorrhoid patients were treated with circular stapler. Whether excisional mucosa stoma ring was regular or not, and its width were recorded. After follow-up from one month to 24 months,the outcomes were compared. RESULTS: The mean age of the patients was (46.6+/- 5.1) years. Regular mucosa stoma ring was excited in 303 cases,while irregular ring in 177 cases. Hemorrhoids prolapse occurred in 18 cases of the patients with irregular mucosal ring excited, while only 8 cases of the patients with regular ring excited (P< 0.01). There was significant difference in the incidence of hemorrhoids prolapse between the different groups according to different width of excisional mucosal stoma ring (P< 0.01), and the patients with postoperative hemorrhoids prolapse was lest with the width of excisional mucosal stoma ring more than 3 cm. CONCLUSIONS: The width of excisional mucosal stoma ring is one of the effecting factors of surgical outcomes. It is effective to excite mucosal stoma ring more than 2 cm,and most effective to excite mucosal stoma ring more than 3 cm. PMID- 16721689 TI - [Application of Ligasure vessel sealing system in haemorrhoidectomy]. AB - OBJECTIVE: To investigate the clinical value of Ligasure vessel sealing system (LVSS) in haemorrhoidectomy. METHODS: From December 2002 to November 2003, clinical data of 36 cases undergoing haemorrhoidectomy with LVSS were compared with those of 30 cases undergoing traditional operation,considering visual analogue scale(VAS), hospital time,operation time,hospital expenses,postoperative complication. RESULTS: There were no significant differences in hospital expenses,postoperative complications between the two groups,but the postoperative pain scores of Ligasure haemorrhoidectomy was better than that of traditional operation. The hospital stay and operation time of Ligasure haemorrhoidectomy were shorter than those of the traditional operation. CONCLUSION: Ligasure vessel sealing system has more advantages such as less pain, safety, facility to perform, and a shorter operation time. PMID- 16721690 TI - [Endoscopic and histopathological features of serrated adenoma of large intestine:an analysis of 71 cases]. AB - OBJECTIVE: To explore the endoscopic and histopathological morphology of large intestinal serrated adenomas (SA). METHODS: The endoscopic and pathological data of 71 SA patients, diagnosed in the Digestive Endoscopy Center, Nanfang Hospital from January 2002 to July 2005, were analyzed retrospectively. RESULTS: Forty seven of the 71 serrated adenomas were protruded (sessile 23, semipedunculated 5, pedunculated 23) and 24 were superficial (flat 16, laterally spreading 8). The mean sizes of the protruded and superficial SA were 10.5 mm and 16.6 mm, respectively, and both of them were frequently located in the sigmoid and rectum. Histopathologically, SA contained tubular glands in 53, tubulovillous glands in 9 and villous glands in 9 cases. Mild dysplasia was found in 47 SAs, moderate dysplasia in 22 SAs, and canceration foci in 2 SAs. The dysplasia of SAs (<10 mm) was significantly better than that of SAs (>or= 10 mm) (P< 0.01). Most IV and III L pit SAs presented villous glands (64%) and tubular glands (68%), respectively. 40% of hyperplastic polyps-like SAs, composed of tubular glands,showed II pit pattern. Atypia in II pit SAs was similar to that in IIIL pit SAs, but was worse than that in IV pit SAs. CONCLUSION: Polyps with II pit pattern possibly are SAs sometimes. SA with the common characters of neoplastic polyps,should be regarded as a new potential precancerous lesion. PMID- 16721691 TI - [Identification of biomarkers in the serum of the patients with poorly differentiated gastric cancer]. AB - OBJECTIVE: To investigate the application of proteomics in the field of serology,and to screen the differential expression proteins related with poorly differentiated gastric carcinoma. METHODS: Two-dimensional electrophoresis (2-DE) was applied to segregate the total proteins in the serum form gastric cancer patients and health volunteers. After staining,the differential expression proteins were analyzed using PDQuest software,and identified using matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). RESULTS: Electrophoresis figures with high resolution and reproducibility were obtained. Six differential expression proteins were found only in the serum from gastric cancer patients, while four other proteins from healthy volunteers. CONCLUSIONS: Protein expression is differential in the serum from the gastric cancer patients and health volunteers. It is hopeful to find the biomarkers related with poorly differentiated gastric carcinoma using proteomics. PMID- 16721692 TI - [Anatomic study of small intestinal vessel in piglet small intestine transplantation]. AB - OBJECTIVE: To study the anatomy of the small intestine,and investigate the optimal selection of donors,recipients,and their small intestine vessels in piglet small intestine transplantation. METHODS: The weight and length of 30 piglets were measured. Angiography and pigments perfusion were used to observe the main vessels of the small intestine,and the length of the small intestine,and the external diameter of the main vessels of the small intestine were measured in vivo and ex vivo. RESULTS: The length of the small intestine was 11.5 times as long as the body length, and its weight accounted for 2.3% of the body weight. The outer diameters of abdominal aorta (AT), mesenteric anterior artery (MAA) and its 5(th)-6(th) branches in vivo and ex vitro were 4.3/4.6mm, 2.5/2.7mm and 1.9/2.2mm respectively. The total number of MAA's branches was 6-8 in general and its 5(th)-6(th) branches were the longest [(20.0 +/- 7.0) mm, (22.0 +/- 8.2) mm]. The outer diameter of mesenterial anterior vein (MAV) was 1-2 mm wider than that of MAA. CONCLUSIONS: AT, MAA and its 5(th)-6(th) branches are the preferable vessels for small intestine transplantation. In segmental small intestine transplantation, the length of the small intestine and body weight can be used to primarily select the suitable animals. PMID- 16721693 TI - [Transcription regulation of 5-Aza-2'-deoxycytidine on maspin gene demethylation in RKO human colorectal cell line]. AB - OBJECTIVE: To detect the methylation status of 5'CpG island in the core promotor of maspin gene in RKO human colorectal cell line,and to explore the transcription regulation of DNA 5'CpG island demethylation on maspin tumor suppressor gene and its effect on the growth of cancer cell. METHODS: The status of 5 'CpG island methylation of maspin gene in RKO human colorectal cell line was analyzed using methylation specific polymerase chain reaction (MSP). After treated with a specific demethylating agent, 5-Aza-2'-deoxycytidine, reverse transcription polymerase chain reaction (RT- PCR) was used to examine maspin gene expression. Cell proliferation was evaluated using MTT assay,distribution of cell cycle and rate of apoptosis were determined using flow cytometry. RESULTS: The 5'CpG island methylation in the core promotor of maspin gene was detected in RKO human colorectal cell line. After treatment with three different concentration of 5-aza 2'-deoxycytidine, the expression of maspin mRNA increased 10.89, 16.91, 23.97 times respectively. MTT array showed the proliferation activity of RKO cell line was obviously reduced after 5-aza-2'-deoxycytidine treatment. The cells were arrested in G(0)/G(1) phase,and the apoptosis rates were 5.17%, 8.71% and 11.23% respectively compared with control group. CONCLUSION: The 5'CpG island methylation is probably responsible for maspin expression silencing in RKO human colorectal cell line, 5-aza-2'-deoxycytidine may effectively cause demethylation and inhibit the growth of tumor cell by reactivating the gene transcription silenced by aberrant hypermethylation. PMID- 16721694 TI - [Expression of vascular endothelial growth factor C and survivin in gastric carcinoma and their clinical implications]. AB - OBJECTIVE: To investigate the expression of vascular endothelial growth factor C (VEGF-C) and survivin protein in human gastric carcinoma,and to evaluate their clinical implications. METHODS: The expressions of VEGF-C and survivin protein in tumor tissues,matched para- tumor tissues from 97 cases with gastric cancer and normal tissues form 20 normal controls,were determined by immunohistochemistry. Their relationships with clinicopathological parameters were analyzed. RESULTS: The positive rate of VEGF-C and survivin protein in tumor tissues (66.0% and 57.2%) was significantly higher than those in matched para-tumor tissues normal tissues (P< 0.05). There were no significant differences in VEGF-C expression considering tumor size,localization,histological grade,venous invasion,and distant metastasis (P > 0.05), while its expression was correlated with serosal infiltration, lymphatic invasion, lymph node metastasis and TNM stage III-IV (P< 0.05). The survivin expression was significantly related with serosal infiltration,lymphatic invasion, regional lymph node metastasis,distant metastasis, and TNM stage III- IV (P< 0.05), but not with histological grade, localization,venous invasion,and tumor size (P > 0.05). The 1, 3 and 5-year survival rates of the patients with positive VEGF-C or survivin protein were significantly lower than those of the patients with negative VEGF-C or survivin (P< 0.05), respectively. In additional,the expression of VEGF-C was positively correlated with survivin expression in gastric carcinoma (P< 0.01). CONCLUSION: The expressions of VEGF-C and/or survivin may be indicators for poor prognosis of gastric carcinoma. PMID- 16721695 TI - Performance of the Italian version of the subjective well-being under neuroleptic (SWN) scale in schizophrenic outpatients. AB - BACKGROUND: Several studies indicate that subjective experience toward antipsychotic drugs (APs) in schizophrenic patients is a key factor in ensuring a smooth recovery from the illness. OBJECTIVE: The principal aim of this study was to establish the psychometric performance of the Subjective Well-being Under Neuroleptic (SWN) scale in its Italian version and to assess, through the SWN scale, the subjective experience of stabilized psychotic outpatients in maintenance with APs. METHODS: The original short version of SWN, consisting of 20 items, was back translated, and a focus group was also conducted to better improve the comprehension of the scale. RESULTS: The results showed a good performance of the Italian version of the SWN as documented by the internal consistency (Cronbach's alpha; 0.85). A satisfactory subjective experience was reported in the sample of schizophrenic outpatients interviewed (SWN mean total score: 84.95, SD: 17.5). CONCLUSIONS: The performance of the SWN scale in the present study was very similar to that reported by Naber et al. in the original validation study. Large multi-center studies are needed to better establish differences in the subjective experience of schizophrenic patients treated with first- and second-generation APs. PMID- 16721696 TI - Fatal poisoning in methadone and buprenorphine treated patients -- are there differences? AB - BACKGROUND: Some recent studies have suggested a lower risk of fatal intoxications in drug-dependent patients under buprenorphine compared to methadone treatment. METHODS: Epidemiological reference data for the Munich region suggest that in 2003 approximately 10 % of all substitution patients were treated with buprenorphine, and 87 % with methadone. We studied the proportion of patients under methadone and buprenorphine substitution among drug-related deaths. Data from forensic post-mortem and toxicological analysis were analyzed. RESULTS: Data indicate that in 96 (35 %) of all 272 so-called drug deaths, methadone was involved compared to a single case of buprenorphine, possibly indicating a relatively better risk profile of buprenorphine. DISCUSSION: More prospective studies are necessary to assess the risk of fatal intoxications under different substitution regimens. PMID- 16721697 TI - The comparison of changes in behavior, neurochemistry, endocrine, and immune functions after different routes, doses and durations of administrations of IL 1beta in rats. AB - BACKGROUND: Increased interleukin-1beta (IL-1) in the brain and periphery has been associated with neurodegenerative and psychiatric disorders. However, results from different IL-1 sources, administrating routes, doses and treatment duration were inconsistent and confused. The neuroendocrine-immune mechanism by which IL-1-induced behavioral changes occur is still unclear. METHODS: In the present study, the acute and sub-chronic effects of rat recombinant IL-1, following either intraperitoneal (ip) or intracerebroventricular (icv) injection, were studied on the behavior, corticosterone secretion, peripheral inflammatory responses and brain monoamines. RESULTS: In the open field apparatus, IL-1 (ip) increased locomotor activity but decreased the activity following icv administration. IL-1 had a greater anxiogenic effect in the elevated plus maze after icv than after ip administration. In the Morris water maze spatial memory was only impaired following sub-chronic and icv administration. Both acute and sub-chronic IL-1 increased the serum corticosterone concentration and decreased the release of the anti-inflammatory cytokine IL-10 from whole blood cultures. However, centrally administered IL-1 increased, while peripherally administered decreased, the release of PGE2 from blood cultures. After sub-chronic administration, the noradrenaline concentration was decreased in several limbic regions, while the turnovers of serotonin and dopamine were increased. DISCUSSION: These results suggest that 1) IL-1 effects depended on the dose, route and duration of administration, and 2) IL-1 enhances the responsiveness of rats to stressful environmental stimuli. In addition, the sub-chronic administration of IL-1 induces behavioral, neurotransmitter, hormonal and immune changes that may be causally implicated in the mechanism of some of psychiatric disorders such as depression. PMID- 16721698 TI - Catecholamine response to methamphetamine is related to glucocorticoid levels but not to pleasurable subjective response. AB - INTRODUCTION: Corticosteroids may modulate addiction. We previously described subjective, physiological, and endocrine effects of 0.5 mg/kg of intravenous methamphetamine after augmenting cortisol level with hydrocortisone or blocking cortisol response with the corticosteroid synthesis inhibitor metyrapone in a double-blind, balanced crossover study. Although the pharmacologic manipulations produced the expected hormonal changes, pleasurable subjective effects of methamphetamine were unchanged. Metyrapone was followed by frequent premature ventricular complexes (PVCs) in two subjects during methamphetamine administration. In order to better understand these results, we examined changes in two plasma catecholamine metabolites, homovanillic acid (HVA) and 3-methoxy-4 hydroxyphenylglycol (MHPG), and their relationship to the previously reported hormonal changes and physiological and subjective responses. METHODS: Plasma from 10 methamphetamine subjects from the earlier study was assayed for HVA and MHPG by high performance liquid chromatography. RESULTS: HVA levels were greater after hydrocortisone or metyrapone pretreatment compared to placebo, and MHPG levels were greater after metyrapone pretreatment. Hydrocortisone pretreatment diminished HVA and MHPG increases after methamphetamine (perhaps explaining the lack of expected increase in pleasurable effects), but metyrapone did not. HVA and MHPG concentrations were not correlated with pleasurable drug effects but were inversely related to reports of "Bad Drug Effect." Increases in MHPG and DHEA concentrations were positively correlated. Metyrapone pre-treated subjects with PVCs had lower HVA and MHPG concentrations. CONCLUSION: Raising cortisol concentration and blocking cortisol synthesis did not produce opposite effects, perhaps because of metyrapone's effect on the hypothalamic-pituitary-adrenal axis, its stress-like effects, and its effects on neurosteroids. PMID- 16721699 TI - Duloxetine treatment of major depressive episodes in the course of psychotic disorders. AB - During the course of psychotic disorders, patients often suffer from intercurrent major depressive episodes (MDEs), and suicides frequently occur. This constellation challenges further improvements in psychopharmacological therapy. The antidepressant duloxetine was recently introduced as a novel reuptake inhibitor of serotonin and noradrenaline. We provide the first reports on duloxetine treatment of MDEs in the course of psychotic disorders. In two cases this substance was successfully involved as an add-on to antipsychotic treatment consisting of clozapine or amisulpride. We achieved a response of the MDEs, as reflected by psychopathological rating scales. A significant rise in the clozapine serum level was detected, most likely because of pharmacokinetic interactions. Overall, the application of duloxetine was well tolerated; therefore, further investigations in prospective studies seem to be recommendable. PMID- 16721700 TI - Aripiprazole pharmacotherapy of borderline personality disorder. AB - We report three consecutive cases of women with borderline personality disorder with psychotic symptoms, who received pharmacotherapy with the new atypical antipsychotic drug aripiprazole. Therapeutic effects were measured using the SCL 90R (symptom check list) and the BSL (borderline symptom list). We observed different responses to aripiprazole. In the first patient we had to discontinue the drug before we were able to observe any therapeutic effects. The second patient also complained about initial side effects. However, after the dose was lowered, the drug was tolerated and she responded well to aripiprazole with respect to all psychopathological aspects. The third patient did not suffer from any side effects under aripiprazole. She responded partially to the drug. Aripiprazole may have a potential role in the pharmacotherapy of borderline personality disorder and may not only target psychotic symptoms in these patients. PMID- 16721701 TI - Effective treatment of coprophagia in a patient with schizophrenia with the novel atypical antipsychotic drug perospirone. AB - Here we report on a patient with schizophrenia who suffered from medication refractory coprophagia. Although there were few cases in which psychotropic medication was effective against coprophagia, we encountered a patient with schizophrenia in whom coprophagia rapidly disappeared after treatment with perospirone, a novel atypical antipsychotic drug of the serotonin-dopamine antagonist (SDA) type. Perospirone has a uniquely high affinity for serotonin-1A receptors, and it could be speculated that perospirone, as a serotonin-1A receptor agonist combined with SDA, may have greater efficacy for treatment refractory symptoms of schizophrenia. Thus, perospirone is an agent with possible efficacy for medication-refractory schizophrenia. PMID- 16721702 TI - Dramatic remission of treatment-resistant depression after the cessation of tricyclic antidepressants. AB - Recently, tricyclic antidepressants (TCAs) have been used mainly for treatment resistant depression (TRD) because of their significant side effects . We report a patient whose prolonged depressive symptoms dramatically improved after the cessation of TCAs. TCAs may cause deterioration of depressive symptoms due to their neurotoxicity. PMID- 16721703 TI - Pergolide as adjuvant therapy to amisulpride in the treatment of negative and depressive symptoms in schizophrenia. AB - Negative and cognitive symptoms of schizophrenia are associated with a hypodopaminergic state in the frontal cortex and do not respond to neuroleptics equally well as positive symptoms. Therefore pharmacological strategies, which increase dopamine metabolism in the mesocortical pathways, may prove beneficial to ameliorate these symptoms. We report on a case of a patient with paranoid schizophrenia, who still presented negative and depressive symptoms during treatment with amisulpride for more than 6 weeks. We prescribed pergolide (a mixed D1/D2 agonist) as adjuvant therapy to treat these symptoms. The patient showed an improvement of global psychopathology, decrease of negative and depressive symptoms, while no changes in positive symptoms nor EPS were present. For this patient, the adjuvant therapy of pergolide to amisulpride constituted a valid pharmacological approach to treat negative and depressive symptoms of schizophrenia, without increasing positive symptoms. PMID- 16721704 TI - Injectable atypical antipsychotics for agitation in borderline personality disorder. AB - Agitation is relatively common among Borderline Personality Disorder (BPD) patients in Psychiatric Emergency Services (PES). New injectable atypical antipsychotics are indicated for treatment in agitated psychotic or maniac patients but not for agitated BDP patients. Twenty agitated BPD patients were treated with intramuscular atypical antipsychotics (olanzapine or ziprasidone). Results suggest intramuscular atypical antipsychotics may be effective, fast and safe for treating acute BPD patients. PMID- 16721705 TI - Interseizure EEG Slowing after ECT is not NCSE. PMID- 16721707 TI - [Parental bonding in childhood and psychological symptoms in a sample of adults with intersexuality]. AB - In a sample of 37 persons with intersexuality the recalled parental bonding and its associations with symptomatic distress in adulthood were analysed. The Questionnaire on Parental Attitudes and Rearing Practices (FEPS), an extended version of the widely used Parental Bonding Instrument, was used to retrospectively assess relevant aspects of perceived parental rearing behaviour. The Brief Symptom Inventory was used to assess actual symptomatic distress. Compared to a community based sample of women, the persons with intersexuality rated their parents as being less caring and showing less emotional warmth while being more punitive. Subsequently, the BSI-cut-off score was used to differentiate clinically disturbed from non-disturbed persons with intersexuality. Analyses revealed that the perceived parental bonding of the non disturbed subgroup was comparable to the community based sample while only the disturbed subgroup differed as described. Using the FEPS-subscales CARE and Punishment Behavior as predictors in logistic regression correctly classified 71.4 % of all subjects as either being clinically disturbed or non-disturbed. PMID- 16721708 TI - [Guideline conformance for outpatient management of COPD in Germany]. AB - BACKGROUND AND OBJECTIVE: Primary care physicians (PCPs) are the ones mainly responsible for the initial diagnosis and outpatient care of patients with COPD. The aim of the present survey was to investigate their initial management of COPD in Germany based on current guidelines and to identify any deviations. METHODS: A prospective cross-sectional survey was conducted as a multiple-choice questionnaire sent out to 1836 PCPs in seven Federal States of Germany (one large town and surrounding country in each). The product-neutral questions focused on the key aspects of current national and international (GLOBAL) COPD guidelines. RESULTS: 486 physicians participated in the study (response rate 26.5%). 66.5% of the physicians used the German COPD guidelines, 20.8% used GOLD guidelines, and only 11.7% observed no guidelines. The physicians were aware of the epidemiological and public health significance of COPD. 76.5% saw spirometry as the diagnostic standard: it was available in 90.1% of the practices. However, only 60-65% were able to cite the correct spirometric criteria for classifying severity of the disease. Educational measures to help patients quit smoking and the teaching and monitoring of patients' inhalation technique were inadequately implemented. The two most important therapeutic goals cited were to improve quality of life (69.1%) and prevent exacerbations (53.1%). Except for the criteria for the use of steroids and the implementation of pulmonary rehabilitation measures, treatment of COPD based on severity class was largely in compliance with the guidelines. However, a significant percentage of the physicians incorrectly assessed the evidence-based clinical benefits of various therapeutic measures. CONCLUSION: The study shows that, despite the high regard in which COPD guidelines are held, deficiencies exist with regard to the diagnosis and treatment of COPD and the practical implementation of educational measures. PMID- 16721709 TI - [Acanthosis nigricans, papillomatosis mucosae and "tripe palms" in a patient with metastasized gastric carcinoma]. AB - HISTORY AND CLINICAL FINDINGS: A 48-year-old obese man presented with thickening, coarseness and hyperpigmentation of the skin, especially of the intertriginous areas, papillomatous to verrucous lesions of the lips and buccal oral mucosa, and hyperkeratosis of the palms ("tripe palms") and soles. He was obese, reported sleep apnea and had a history of hyperuricemia, mixed hyperlipidemia and previous myocardial infarction. He was on a maintenance dose of a proton pump inhibitor for chronic gastro-esophageal reflux. EXAMINATIONS: Immunohistochemical studies of the skin lesion revealed increased epidermal immunoreactivity for the melanocortin-1-receptor. Increased levels of tumor markers CA 19-9 (141100 U/ml), CA 72-4 (755 U/ml) and CEA (189 ng/ml) were found in the serum. Gastroscopic findings were suspicious of adenocarcinoma of the stomach: it was classified histologically as a signet-ring cell, non-mucinous adenocarcinoma. At the time of diagnosis the tumor had already metastasized to perigastric and peripancreatic lymph nodes with peritoneal carcinosis. TREATMENT AND COURSE: Since a curative resection was impossible a gastrojejunostomy was carried out. After this the patient received several courses of chemotherapy according to different schemes. Serum tumor marker levels and cutaneous signs regressed several times. CONCLUSIONS: Marked acanthosis nigricans -- especially when associated with further cutaneous markers of malignancy, e.g. mucocutaneous papillomatosis or so called tripe palms -- calls for thorough search for malignant tumor, also if metabolic or endocrinological abnormalities co-exist. A pathogenetic role of a melanocyte-stimulating hormone in the development of the skin changes is suggested. PMID- 16721710 TI - [Acute hepatitis due to kava-kava and St John's Wort: an immune-mediated mechanism?]. AB - HISTORY AND CLINICAL FINDINGS: In an otherwise healthy 48-year-old female patient, acute hepatitis with transaminase increase (GOT up to 613 U/l, GPT up to 752 U/l), inconspicuous hepatitis serology findings, negative autoantibody status and negative virus serology was observed after a 10-week long intake of kava-kava (1-3 x 200 mg/day) and St John's Wort (1 x 425 mg/day). Biopsy of the liver showed lobular and portal necroinflammatory activity without indication of cirrhosis. DIAGNOSIS: Due to these findings with proven T-cell activity (lymphocyte typing, neopterin determination) as well as the aetiopathology, this form of hepatitis with histological characteristics of a nutritive/medicinal toxic origin was classified as induced immunologic idiosyncratic hepatitis, possibly in terms of an antibody-negative autoimmune hepatitis. TREATMENT AND CLINICAL COURSE: Discontinuation of the existing medication and simultaneous onset of immunosuppressive combination therapy of cortisone, azathioprine and ursodeoxycholic acid resulted in normalisation of the liver parameters within a period of two months. CONCLUSION: On the one hand, it appears that simultaneous intake of St John's Wort possibly potentiates the toxicity of kavapyrones. On the other hand, an immune-mediated mechanism, induced by kava-kava, cannot be completely excluded in the present case. It must be stressed that in patients with autoimmune hepatitis, precise history of medication intake should also be available. PMID- 16721711 TI - [Tiotropium]. PMID- 16721713 TI - [Raynaud's phenomenon--current diagnosis and therapy]. PMID- 16721714 TI - [Diagnosis of myocarditis: description and assessment of available methods]. AB - Myocarditis is a common disease in all age groups. Correct diagnosis of myocarditis is difficult, but of clinical and scientific importance. It is the aim of this review to describe and evaluate diagnostic possibilities. It is not possible to make a definitive diagnosis of myocarditis by electrocardiography, laboratory tests or echocardiography. Nor can techniques of nuclear medicine or coronary angiography provide an unequivocal diagnosis of myocarditis. Myocarditis can be diagnosed with certainty only by endomyocardial biopsy, which must be examined by histological, immunohistological and molecular techniques to obtain maximal sensitivity. But optimal diagnosis of a biopsy is subject to the problem of sampling error. Furthermore, myocardial biopsy is in practice used only rarely, despite a low complication rate. Thus the diagnosis of myocarditis is often based merely on suspicion. The limitations of standard methods employed so far have made cardiac magnetic resonance imaging (CMR) a valuable new additional test when there is the clinical suspicion of myocarditis. A myocardial biopsy will often reveal myocarditis in patients with cardiac symptoms and CMRI findings typical of myocarditis, especially if the biopsy was obtained near the area of contrast enhancement in the CMRI. CMRI can also be used for noninvasive and low risk follow-up of functional parameters and tissue damage. PMID- 16721715 TI - [Drug-releasing stents. Little use in cardiology despite excellent data--what next?]. PMID- 16721717 TI - [Comment on the article, "ENT relevant changes in the operation and procedural classification key starting 2006" by J. Alberty and D. Franz]. PMID- 16721718 TI - [Nasal irrigation in case of rhinosinusitis]. AB - Nasal irrigation has proven in several recently published articles its clinical effectiveness and its neglectable rate of side effects even in children and during pregnancy. In most guidelines for the treatment of various nasal pathological conditions nasal irrigation is recommended as stand alone or as additional therapy. Although no common treatment protocol exists, a isotonic (0.9 %) or slightly hypertonic solution (3 - 5 %) is suggested because of its positive influence upon mucociliary clearance. Buffered solutions bring a more pleasant feeling during rinsing, which explains also the beneficial effect of natural brine solutions. In children, nasal irrigation can save antibiotics and reduce decongestant to nil. Important is the hygienic application in order not to introduce additional pathogenic germs into the nasal system. PMID- 16721719 TI - Structural analysis of the unique insecticidal activity of novel mungbean defensin VrD1 reveals possibility of homoplasy evolution between plant defensins and scorpion neurotoxins. AB - A variety of evolutionarily related defensin molecules is found in plants and animals. Plant gamma-thionins and scorpion neurotoxins, for instance, may be categorized in this functional group, although each class recognizes a distinct receptor binding site. Such molecules are also categorized into the superfamily of cysteine-rich proteins. Plant defensins were generally believed to be involved in antimicrobial or antifungal mechanisms and, unlike scorpion toxins, little is known about whether these molecules are also endowed with the function of insect resistance. We have previously reported the isolation of a cDNA encoding a small cysteine-rich protein designated VrD1 (VrCRP) from a bruchid-resistant mungbean, which is apparently the first discovered plant defensin exhibiting in vitro and in vivo both insecticidal and antifungal activities. Our previous data also successfully demonstrated that VrD1 is toxic to E. coli and able to completely arrest the growth of Sf-21 insect cells at low concentration. However, the molecular and structural basis of this unique insecticidal activity of VrD1 is not clear. Therefore, in the present study, we use structural approach and phylogenic analysis to investigate the evolutionary and functional relations for such unique insecticidal activity. From our results, it is suggested that VrD1, in addition to gamma-thionins and several amylase inhibitors, is highly homologous to scorpion toxins, especially the short toxins. Moreover, based on the observation from our homology structures, VrD1 may utilize a newly found cluster of basic residues to achieve its insecticidal function, whereas all the other plant gamma-thionins were known to use a previously identified basic cluster conserved for gamma-thionins. Considering the general feature of short scorpion toxins to act on insect cell membranes with K(+)- or Cl(-)-channels as molecular targets, our analysis of interaction and recognition modes provides reasonable correlations between this newly found basic cluster and the insecticidal activity of VrD1, which is also comprehended as a possible link for "homoplasy evolution" between plant and animal defensin molecules. PMID- 16721720 TI - Multiple pathways in conformational transitions of the alanine dipeptide: an application of dynamic importance sampling. AB - We present multiple dynamic transition pathways on the two-dimensional dihedral plane between conformational states of the alanine dipeptide. The method used in this study is dynamic importance sampling (DIMS). To perform DIMS, unbiased molecular dynamic simulations are used to generate equilibrium ensembles for the alanine dipeptide within different states. Free energy surfaces on the dihedral plane are calculated from the equilibrium simulations, and four energy minima defined from the surface are used as the starting and ending points for DIMS dynamics. The DIMS method represents an important step towards finding multiple transition pathways within complex biomolecular systems. PMID- 16721721 TI - Accurate prediction of the blood-brain partitioning of a large set of solutes using ab initio calculations and genetic neural network modeling. AB - A genetic algorithm-based artificial neural network model has been developed for the accurate prediction of the blood-brain barrier partitioning (in logBB scale) of chemicals. A data set of 123 logBB (115 old molecules and 8 new molecules) of a diverse set of chemicals was chosen in this study. The optimum 3D geometry of the molecules was estimated by the ab initio calculations at the level of RHF/STO 3G, and consequently, different electronic descriptors were calculated for each molecule. Indeed, logP as a measure of hydrophobicity and different topological indices were also calculated. A three-layered artificial neural network with backpropagation of an error-learning algorithm was employed to process the nonlinear relationship between the calculated descriptors and logBB data. Genetic algorithm was used as a feature selection method to select the most relevant set of descriptors as the input of the network. Modeling of the logBB data by the only quantum descriptors produced a 5:4:1 ANN structure with RMS error of validation and crossvalidation equal to 0.224 and 0.227, respectively. Better nonlinear model (RMS(V) and RMS(CV) equals to 0.097 and 0.099, respectively) was obtained by the incorporation of the logP and the principal components of the topological indices to electronic descriptors. The ultimate performances of the models were obtained by the application of the models to predict the logBB of 23 molecules that did not have contribution in the steps of model development. The best model produced RMS error of prediction 0.140, and could predict about 98% of variances in the logBB data. PMID- 16721722 TI - Discordant anatomically corrected malposition of the great vessels and Ebstein's malformation: fetal and neonatal findings. AB - Discordant anatomically corrected malposition of the great vessels (ACM) is a rare congenital cardiac malformation that is often associated with other cardiac lesions. To our knowledge, a fetal diagnosis of this lesion has not yet been reported in the literature. We present a case of ACM associated with Ebstein's malformation, diagnosed by fetal echocardiography. PMID- 16721723 TI - Fetal atrioventricular valve junction in normal fetuses and in fetuses with complete atrioventricular septal defect assessed by 4D volume rendering. AB - OBJECTIVES: To assess the feasibility and clinical potential of 4D volume rendering of the atrioventricular (AV) valve junction, to standardize the acquisition method, and to display the AV valve junction morphology in normal fetuses and in those with a complete atrioventricular septal defect (AVSD). METHODS: We performed sonography in 40 normal fetuses and 10 fetuses with complete AVSD, and volume datasets were acquired from apical and lateral four chamber views. The render box was placed systematically. First, it included the AV valves, with the reference dot at the level of the crux of the heart. Then, it included the papillary muscles, with the reference dot in the interventricular septum at the level of the distal opening of the tricuspid valve leaflet. RESULTS: Volume acquisition and rendering were technically possible in all cases. Volume rendering of the left ventricle showed the position of the anterolateral and posteromedial papillary muscles in 36/40 normal fetuses (90%). At the level of the right ventricle, the septal, anterior and posterior papillary muscles were visualized in 33/40 normal fetuses (82%). In cases of complete AVSD, the AV valve has five leaflets, with anterosuperior and posteroinferior bridging leaflets straddling the septa. The morphology of the anterosuperior bridging leaflets and the abnormal position of the papillary muscles could be displayed in all cases. CONCLUSIONS: Our study suggests that some of the components of the AV junction can be reconstructed easily from sonographic volumes acquired from an apical or lateral four-chamber view. This new technique may have a role in obtaining views that are not easily accessible by standard sonography, enabling a rapid complementary assessment of normal and abnormal intracardiac anatomy. PMID- 16721724 TI - 2D representation of protein secondary structure sequences and its applications. AB - In terms of the classification of the protein secondary structures, we propose a 2D representation of protein secondary structure sequences. The representation are used to display, analyze, and compare the secondary structure sequences. Based on this representation, we assign the structural class to the protein, and verify the advantage or disadvantage of the methods of predicted protein second structure. PMID- 16721725 TI - Tobacco smoking, NAT2 acetylation genotype and breast cancer risk. AB - The role of active and passive cigarette smoking in breast cancer etiology remains controversial. Using data from a large population-based case-control study in Poland (2386 cases, 2502 controls) conducted during 2000-2003, we examined the associations between active and passive smoking overall and for different age categories. We also evaluated differences in risk by estrogen receptor (ER) and progesterone receptor (PR) status in tumors, and the potential modification of the smoking association by N-acetyl transferase 2 (NAT2) genotype. Women ever exposed to passive smoking at home or at work had a risk of breast cancer similar to those never exposed to active or passive smoking (OR (95%CI) = 1.11 (0.85-1.46), and no trends were observed with increasing hours/day years of passive smoking exposure. Active smoking was associated with a significant increase in risk only among women younger than 45 years of age (OR (95%CI) = 1.95 (1.38-2.76); 1.15 (0.93-1.40); 0.91 (0.77-1.09) for < 45, 45-55 and > 55 years of age, respectively; p-heterogeneity < 0.001 for < 45 vs. > 55 years) and prevailed for both ER+ and ER- tumors. The smoking association among women < 45 years was stronger for current than former smokers, and a significant trend was observed with duration of smoking (p = 0.04). NAT2 slow vs. rapid/intermediate acetylation genotype was not related to breast cancer risk (0.99 (0.87-1.13)), and did not significantly modify the smoking relationships. In conclusion, our data indicate that passive smoking is not associated with breast cancer risk; however, active smoking might be associated with an increased risk for early onset breast cancers. PMID- 16721726 TI - Identification of a novel amplicon at distal 17q containing the BIRC5/SURVIVIN gene in malignant peripheral nerve sheath tumours. AB - Previous studies have suggested that amplification of genes, notably the TOP2A gene, on chromosome arm 17q may be important for the development of malignant peripheral nerve sheath tumour (MPNST). In order to study the frequency, distribution, and chromosomal organization of rearrangements at 17q, interphase and metaphase fluorescence in situ hybridization (FISH) were used to evaluate copy number changes at 17q in 28 MPNSTs. Increased copy numbers were seen for the ERBB2 and TOP2A genes in eight and nine cases, respectively, supporting a potential role for these two genes in MPNST tumourigenesis. Net gain of distal 17q material was observed in 16 of the 28 MPNSTs, with high-level gain in three cases, and was associated with poor outcome. Among the 26 patients for whom follow-up data were available, gain of distal 17q was present in 11 of 12 tumours that had metastasized, compared with 4 of 14 of those that had not metastasized. Detailed FISH mapping analysis of metaphase spreads identified a 2 Mb commonly gained/amplified region at 17q25. Among the genes mapping to this region, BIRC5, which encodes the baculoviral IAP repeat-containing protein 5/survivin protein, is a strong candidate target gene for amplification, as it has been previously shown to be overexpressed in neurofibromatosis type 1-associated MPNST. Three other genes that co-amplified with BIRC5 represent other potential candidate genes: PTDSR involved in apoptosis; SEPT9 overexpressed in human malignant brain tumours; and SOCS3 involved in cell survival and differentiation of neurons. PMID- 16721727 TI - Interconversion and tissue distribution of pentoxifylline and lisofylline in mice. AB - The aim of this study was to assess the interconversion pharmacokinetics and tissue distribution of pentoxifylline and the active (R)-enantiomer of its metabolite M1, lisofylline in male CD-1 mice. Both compounds were administered intravenously at a dose of 50 mg/kg on two separate occasions. Serum and tissues were collected at different time points following drug administration. In addition, the (S)-enantiomer of M1 was administered to a group of mice and serum samples were obtained. Analyte concentrations were measured by chiral HPLC. All serum concentration versus time data were fitted simultaneously to a pharmacokinetic model incorporating interconversion processes of parent drug and metabolites. The estimated conversion clearance of (-)-(R)-M1 to pentoxifylline (CL21) was six times greater than that for the reverse process (CL12). The interconversion of pentoxifylline and (+)-(S)-M1 was faster as reflected by the values of conversion clearances CL13 and CL31 which were approximately 16 and 7 times greater in comparison with the corresponding clearances for the interconversion of pentoxifylline and (-)-(R)-M1. When fitting pharmacokinetic data of both parent compounds to a one-compartment model, the values of elimination clearances assessed were close to those obtained on the basis of the interconversion model. After administration of pentoxifylline, tissue-to-serum AUC ratios ranged from 0.1 for liver and lungs to 0.32 for brain tissue. Serum levels of its metabolite, (-)-(R)-M1 were very low, whereas its tissue levels exceeded serum concentrations. The highest value of metabolite-to-parent AUC ratio (4.98) was observed in lungs. When (-)-(R)-M1 was given as a parent drug, tissue-to-serum AUC ratios in liver, kidney, and lungs were very close and ranged from 0.64 to 0.72. At the same time, levels of its metabolite, pentoxifylline were relatively low both in serum and all tissues studied. In consequence, metabolite-to-parent AUC ratios did not exceed the value of 0.27. In conclusion, reversible metabolism plays a modest role in the disposition of pentoxifylline and (-)-(R)-M1. It seems that pentoxifylline has less favourable pharmacokinetic properties than (-)-(R)-M1 due to lower concentrations attained in target organs. High levels of (-)-(R)-M1 observed after pentoxifylline administration in certain tissues such as liver or lungs suggest that pentoxifylline may constitute an effective prodrug for (-)-(R)-M1 in these organs. PMID- 16721729 TI - Formation of RACK- and grid-type metallosupramolecular architectures and generation of molecular motion by reversible uncoiling of helical ligand strands. AB - The interaction of appropriate metal ions (Pb(II), Zn(II)) with helical ligand strands, obtained by hydrazone polycondensation, generates polymetallic supramolecular architectures of rack and grid types, by uncoiling of the ligand. The interconversion between the helical free ligand and the linearly extended ligand in the complexes produces reversible ion-induced, nanomechanical molecular motions of large amplitude. It has been integrated in an acid-base neutralisation fuelled process, which links the extension/contraction of the ligand strands to alternating changes in pH. PMID- 16721730 TI - OFF-off rebound dyskinesia in subthalamic nucleus deep brain stimulation of Parkinson's disease. AB - A 61-year-old man with Parkinson's disease (PD), motor fluctuations, and dyskinesias underwent bilateral implantation of deep brain stimulation (DBS) electrodes in the subthalamic nucleus (STN). One month after surgery, DBS was optimized to bilateral monopolar settings at the most proximal electrode just superior to the STN, which improved motor fluctuations and dyskinesias. At several postoperative evaluations off medications overnight, both stimulators were turned off and within 60 seconds he developed severe dyskinesias. When the stimulators were turned back on, the dyskinesias soon resolved. This article is a first report of a unique pattern of rebound-type dyskinesia that occurred in the off medication state produced by stopping STN DBS. PMID- 16721731 TI - Electrophysiological deterioration over time in patients with Huntington's disease. AB - In recent studies aimed at assessing the effects of original therapeutic strategies applied to patients with Huntington's disease (HD), we observed informative changes in electrophysiological results that recovered normal values in coherence with clinical improvement. However, longitudinal studies were lacking for determining whether electrophysiological test results evolve in parallel with clinical markers of the natural course of the disease and could consequently provide objective quantifiable markers of disease progression. For this purpose, electrophysiological testing was performed annually in a cohort of 20 patients with HD over a 2-year period (three examinations). The study included the recording of sympathetic skin responses and blink reflexes (BRs) to supraorbital nerve stimulation, long latency reflexes (LLRs) and somatosensory evoked potentials (SEPs) to median nerve stimulation, and cortical silent periods (CSPs) to transcranial magnetic stimulation. Clinical evaluation was based on the Total Functional Capacity scale (TFC) and the Motor part of the Unified Huntington's Disease Rating Scale (UHDRS). A significant deterioration with time was found for BR latency, LLR presence, various SEP parameters (parietal N20 peak amplitude and frontal N30 presence) and CSP duration. Attenuation of the N20 peak and CSP shortening correlated with functional decline, as assessed by the TFC score, whereas delayed BR and LLR abolition correlated with UHDRS Motor score deterioration. This study shows that several electrophysiological parameters are closely associated with dysfunction of various neural circuits in HD and could be useful markers of disease progression. PMID- 16721732 TI - Subtypes of mild cognitive impairment in Parkinson's disease: progression to dementia. AB - The aim of this study was to establish the rate of progression from mild cognitive impairment (MCI) to dementia in patients with Parkinson's disease (PD). PD patients without dementia were recruited in 1997 from an ongoing prospective epidemiological study. The assessment included neurological and psychiatric examinations, a clinical interview based on the Diagnostic and Statistical Manual of Mental Disorders, Revised Third Edition (DSM-III-R) criteria for dementia, and a battery of neuropsychological tests. PD was diagnosed according to established criteria, dementia was diagnosed according to the DSM-III-R criteria, and subtypes of MCI were classified according to modified Petersen's criteria. Seventy-two nondemented PD patients were included. A total of 34 were cognitively intact, whereas 38 were diagnosed with MCI (amnestic, n = 6; single nonmemory domain, n = 17; multiple domains slightly impaired, n = 15). Fifty-nine patients (82%) completed follow-up examination 4 years later, and 18 (62%) of the patients with MCI and 6 (20%) of the cognitively intact PD patients were demented (P = 0.001). Single domain nonmemory MCI and multiple domains slightly impaired MCI were associated with later development of dementia (P = 0.003; P = 0.04), whereas amnestic MCI subtype was not (P = 0.76). We conclude that patients with PD and MCI had a higher risk of developing dementia than cognitively intact PD patients, suggesting that MCI in PD is an early manifestation of dementia. However, these findings should be interpreted with caution due to the relatively small number of subjects included in this study. PMID- 16721733 TI - Identification of mannose moieties in N- and O-linked oligosaccharides of the primordial germ cells of Xenopus embryos. AB - The presence of mannose (Man) in the glycoconjugates of primordial germ cells (PGCs) of Xenopus embryos was elucidated by lectin histochemistry with Concanavalin A (Con A) and snowdrop (Galanthus nivalis) bulb lectin (GNA), in combination with deglycosylative pretreatments: beta-elimination, which removes O linked oligosaccharides, and incubation with Peptide N glycosidase F (PNGase F), which removes N-linked glycan chains. In addition, histochemistry with Con A, which binds to Man and glucose (Glc), was also performed after glucose-oxidase incubation, which converts Glc into gluconic acid, and GNA was carried out after acid hydrolysis, which removes terminal sialic acid (NeuAc) moieties. PGCs were analyzed during their migration over the mesentery until the genital ridge, and after colonization of this gonad anlage. The results showed that for both lectins: (1) the PGCs and other surrounding tissue showed a similar binding pattern, and (2) the staining in the PGCs was similar in the developmental stages studied. Labeling with Con A was due to Man, and not to Glc, as shown after incubation with glucose-oxidase, and it was assumed that Man was in N-linked oligosaccharides. However, GNA labeling was mainly due to O-linked oligosaccharides, because the pretreatment of beta-elimination turned cells negative. Moreover, acid hydrolysis pretreatment gave rise to a stronger GNA staining, suggesting that either Man was also in subterminal position to NeuAc or some Man-containing glycans were unmasked after removal of NeuAc from other oligosaccharide chains. PMID- 16721734 TI - Immunohistological evidences of Ginkgo biloba extract altering Bax to Bcl-2 expression ratio in the hippocampus and motor cortex of senescence accelerated mice. AB - Ginkgo biloba extract 761 appears to display neuroprotective effect in many nervous diseases and aging. Deterioration of mental functions during aging is always accompanied by loss of neurons, presumably through apoptosis. Here, we studied the effect of G. biloba extract in the expression of Bax/Bcl-2 ratio, an important apoptotic index, in the hippocampus and motor cortex of the aging brain. Bax and Bcl-2 expressions were examined with immunohistochemical methods. Senescence Accelerated Mice Prone Strain 8 was used because the aging process was accelerated with neuropathological alterations similar to those found in the aging human brain. The mice were fed with either G. biloba extract or sucrose from the age of 3 weeks until sacrifice at 3 or 9 months old. In the hippocampus of G. biloba fed 9-month-old mice, the ratio of Bax positive cell to Bcl-2 positive cell (Bax/Bcl-2 expression ratio) was 11.43 +/- 3.11 (mean +/- SD); significantly lower (P < 0.05) than the Bax/Bcl-2 expression ratio of 20.99 +/- 5.34 in the sucrose fed mice. The Bax/Bcl-2 expression cell ratios, however, in the motor cortex were not significantly different between the two groups (2.22 +/ 1.35 versus 2.27 +/- 2.02 for the G. biloba and the sucrose fed mice, respectively). The decrease in the Bax/Bcl-2 expression cell ratio following G. biloba treatment might hence be able to protect the aging hippocampus from moving further down the apoptotic pathway. Western blotting confirmed the decrease of Bax in the brain even after a short term and high dose Ginkgo treatment. It is speculated that the G. biloba extract may be a potential neuroprotective agent against apoptosis through the differential expressions of the Bax and Bcl-2 in the hippocampus. PMID- 16721735 TI - First clinical case of boron neutron capture therapy for head and neck malignancies using 18F-BPA PET. AB - BACKGROUND: We investigated the application of boron neutron capture therapy (BNCT) to suitable cancers other than glioblastoma and melanoma. Head and neck malignancies were consequently selected as adaptable cancers. We reported the clinical results of our first case treated and discussed several advantages to the application of BNCT to head and neck tumors. METHODS: The patient was a 48 year-old woman with recurrent submandibular gland cancer. We confirmed the p boronophenylalanine (BPA)-accumulating capacity of the tumor by fluorine-18 labeled p-boronophenylalanine positron emission tomography ((18)F-BPA PET) before BNCT. The tumor/normal tissue boron concentration ratio was 2.9. The patient underwent a preirradiation CT scan for treatment planning performed using the "SERA" software program. The tumor was irradiated at the Kyoto University Research Reactor with epithermal neutrons 5 MW for 90 minutes. The tumor dose and normal tissue dose calculated ranged from 20.0 to 25.2 Gy and from 3.2 to 5.8 Gy, respectively. RESULTS: To date there has been continuous complete regression in the tumor and no acute and chronic complications for 1.5 years. CONCLUSIONS: Although only 1 patient has shown complete regression and additional long-term follow-up should be required to assess this treatment, we believe that head and neck tumors are suitable for BNCT and that such excellent results will have a great impact on patients in the near future. PMID- 16721736 TI - Comparison of the domains of anxiety and mood of the University of Washington Head and Neck Cancer Questionnaire (UW-QOL V4) with the CES-D and HADS. AB - BACKGROUND: Version 4 of the University of Washington Head and Neck Cancer Questionnaire (UW-QOLv4) includes items on mood and anxiety. The aim of this study was to compare the responses to these single items with the Centre for Epidemiology Studies Depression Scale (CES-D) and the Hospital Anxiety Depression Scale (HADS). METHODS: A cross-sectional postal survey was undertaken in April 2003. The survey was composed of all patients treated for oral and oropharyngeal squamous cell carcinoma between 1992 and 2002 who were alive and disease free. RESULTS: We distributed 306 questionnaires; there were 197 replies (65%) from 110 male and 87 female patients. Most patients reported relatively little depression, with 170 of 190 (89%) reporting a HADS depression score of less than 11. Similarly, most patients were not anxious, with 158 of 183 (86%) reporting a HADS anxiety score of less than 11. UW-QOL mood, UW-QOL anxiety, HADS anxiety, HADS depression, and CES-D scores were all moderately intercorrelated (Spearman correlations from 0.39-0.68 ignoring the signs, all p < .001). The UW-QOL mood correlated with the scores and "case-ness" categories of the HADS depression and CES-D scales, whereas the UW-QOL anxiety correlated with the scores and "case ness" of the HADS anxiety. CONCLUSIONS: Questions on mood and anxiety can help identify significant psychological morbidity, taking a score of less than 75 for UW-QOL mood and less than 70 for UW-QOL anxiety. This could be used to trigger formal psychological assessment and with a view to possible therapeutic intervention. PMID- 16721737 TI - Kaposiform hemangioendothelioma arising in the ethmoid sinus of an 8-year-old girl with severe epistaxis. AB - BACKGROUND: Epistaxis is very common during childhood. It occurs primarily in boys and is usually self-limiting. Trauma and nose picking are among the most common causes. In general, epistaxis can be easily treated with anterior nasal packing or electrocoagulation. METHODS: We report a case of an 8-year-old girl with severe unilateral epistaxis. RESULTS: The bleeding originated from a kaposiform hemangioendothelioma arising in the left nasal cavity and ethmoid sinus. The feeding vessels originating from the maxillary artery were first embolized. The tumor was then surgically removed through a combined external ethmoidectomy and endonasal approach. The postoperative course was uneventful. MRI at 6 months after surgery showed no tumor recurrence. CONCLUSIONS: We report a previously undescribed cause of epistaxis in children, namely, a kaposiform hemangioendothelioma. To our knowledge, this is the first such case in the English-language literature. The differential diagnosis of severe unilateral nasal bleeding among the pediatric population should include the possibility of a kaposiform hemangioendothelioma. PMID- 16721738 TI - Esthesioneuroblastoma recurrence presenting as a syndrome of inappropriate antidiuretic hormone secretion. AB - BACKGROUND: Esthesioneuroblastoma is an uncommon intranasal tumor. These neuroendocrine neoplasms are rarely associated with excess hormone syndromes, and only nine cases of inappropriate antidiuretic hormone syndrome (SIADH) secondary to an esthesioneuroblastoma have been described. In all these cases, electrolyte abnormalities were seen when the tumor initially developed. We report a unique case of esthesioneuroblastoma recurrence manifesting as SIADH as the solely presenting feature. METHODS AND RESULTS: A 34-year-old woman was referred to us for evaluation for hyponatremia. She had undergone resection of an esthesioneuroblastoma at age 18 with radiotherapy. The patient had undergone annual CT of the nasal area and was considered to be disease free. The study of the hyponatremia was consistent with a SIADH, and MRI revealed an intranasal mass. The resection of the tumor reversed the hyponatremia, and pathologic analysis revealed a recurrence of esthesioneuroblastoma. CONCLUSIONS: Biochemical analysis should be performed in the follow-up of patients with esthesioneuroblastoma. In our case, the biochemical abnormality led to the diagnosis of the recurrence. PMID- 16721739 TI - Response to intraarterial induction chemotherapy: a prognostic parameter in oral and oropharyngeal cancer. AB - BACKGROUND: Patients with head and neck cancer and good pathologic response to neoadjuvant systemic induction chemotherapy have a better prognosis for survival than do those with stable or progressive disease. Thus, induction chemotherapy could theoretically help in stratifying further treatment, but toxicity is much too high. The prognostic implication of superselective intraarterial high-dose cisplatin administered by a femoral approach, which has much less toxicity, is not yet known. METHODS: One hundred eighty-seven unselected consecutive patients with previously untreated oral and oropharyngeal squamous cell carcinoma received intraarterial high-dose cisplatin for induction and were assessed for response by visual examination and palpation. This treatment was followed by surgery and adjuvant radiation with concomitant systemic chemotherapy. Omission of a modality depended on individual contraindications and not on preselection. The consequence of omissions has been the constitution of several treatment arms. The overall and disease-free survival in relation to clinical local response after intraarterial induction chemotherapy was calculated using the Kaplan-Meier method. Additional analysis excluded bias caused by stages and treatment arms. RESULTS: Explorative statistics using the log-rank and chi-square tests demonstrated a strong prognostic relevance of response to intraarterial chemotherapy irrespective of stage and treatment. CONCLUSIONS: Our results are encouraging for prospective randomized studies and molecular genetic investigations with intraarterial chemotherapy. PMID- 16721740 TI - Genetic polymorphisms of CYP1A1, CYP2E1, GSTM1, and GSTT1 associated with head and neck cancer. AB - BACKGROUND: Alcohol intake and tobacco smoke, in addition to other environmental and genetic factors, have been associated with head and neck cancer. We evaluated the role of metabolic enzyme polymorphisms on the risk of head and neck cancer in a hospital-based case-control study. METHODS: CYP1A1MspI, CYP2E1PstI, GSTM1, and GSTT1polymorphisms were evaluated in 103 histologically confirmed head and neck cancer cases and 102 controls by means of polymerase chain reaction-restriction fragment length polymorphism methods. RESULTS: GSTM1null increased the risk of head and neck cancer (odds ratio [OR], 2.2; 95% confidence interval [95% CI], 1.24-3.79), oral cancer (OR, 2.8; 95% CI, 1.28-5.98), and pharyngeal cancer (OR, 2.2; 95% CI, 1.08-4.63). CYP2E1PstI polymorphism indicated a risk for oral cancer (OR, 3.6; 95% CI, 1.29-11.56). The joint effect of GSTM1 null and CYP1A1 polymorphism increased the risk of head and neck cancer (OR, 2.4; 95% CI, 1.13 5.10). CONCLUSIONS: GSTM1 null alone or associated with CYP1A1 increased the risk of head and neck cancer; the CYP2E1PstI mutated allele increased the risk for only oral cancer. PMID- 16721741 TI - Induction chemotherapy with cisplatin and gemcitabine followed by accelerated radiotherapy and concurrent cisplatin in patients with stage IV(A-B) nasopharyngeal carcinoma. AB - BACKGROUND: The purpose of this study was to evaluate the efficacy and toxicity of cisplatin plus gemcitabine as induction chemotherapy in advanced nasopharyngeal carcinoma (NPC). METHODS: Thirty-seven patients with stage IV(A-B) NPC were treated with 3 cycles of cisplatin plus gemcitabine (cisplatin 80 mg/m(2) on day 1; gemcitabine 1250 mg/m(2) on days 1 and 8) 3-weekly as induction chemotherapy, followed by another 3 cycles of concurrent cisplatin (100 mg/m(2) on day 1) 3-weekly with accelerated radiotherapy (RT) at 70 Gy in 2-Gy fractions, 6 daily fractions per week. RESULTS: The overall response rate to induction chemotherapy was > 90%, and side effects other than uncomplicated hematologic toxicities were uncommon. All patients completed RT, with 92% receiving > or = 5 cycles of chemotherapy. At a median follow-up of 2.9 years, the 3-year overall survival (OS) and disease-free survival (DFS) rates were 76% and 63%, respectively. CONCLUSIONS: Cisplatin plus gemcitabine is a well-tolerated, effective, and convenient induction chemotherapy regimen and warrants further studies to confirm its benefit in advanced NPC. PMID- 16721742 TI - Interstitial low-dose-rate brachytherapy in the treatment of recurrent head and neck malignancies. AB - BACKGROUND: Recurrent head and neck malignancies are therapeutically challenging. Brachytherapy is a retreatment alternative to external-beam radiation therapy (EBRT). METHODS: Patients receiving brachytherapy during 1987-2004 for recurrent head and neck cancer were identified. Tumor and treatment characteristics and toxicities were recorded. Progression-free survival (PFS) and overall survival (OS) estimates were generated. The influence of prognostic factors was determined. RESULTS: Eighty-two patients were analyzable. Analysis was limited to patients who had brachytherapy for a first recurrence (n = 45). Brachytherapy (> or = 55 Gy) was a monotherapy in 22 of 45 patients. As part of their salvage brachytherapy treatments, 14 patients also underwent surgery; 3 patients also underwent EBRT; and 6 patients underwent surgery, EBRT, and brachytherapy. Retreatment morbidity included acute toxicity (n = 7) and late toxicity (n = 18). Median PFS was 15 months, and locoregional control rates at 1 and 2 years were 50% and 37%, respectively. Time to progression differed by site of the primary tumor (p = .10). Median OS was 16 months, and OS at 2 and 5 years was 33% and 11%, respectively. CONCLUSIONS: Brachytherapy for recurrent head and neck cancer has an acceptable toxicity profile and is viable alternative to EBRT. Further optimization of the best sites and doses for neck brachytherapy is required. PMID- 16721743 TI - Management of contralateral N0 neck in oral cavity squamous cell carcinoma. AB - BACKGROUND: The purpose of this study was to evaluate the incidence and predictive factors of contralateral occult lymph node metastasis in squamous cell carcinomas of the oral cavity to form a rational basis for elective contralateral neck management. METHODS: We performed a retrospective analysis of 66 patients with cancer of the N0-2 oral cavity undergoing elective neck dissection for contralateral clinically negative necks from 1991 to 2003. RESULTS: Clinically negative but pathologically positive contralateral lymph nodes occurred in 11% (7 of 66). Of the 11 cases with a clinically positive ipsilateral node neck, contralateral occult lymph node metastases developed in 36% (4 of 11), in contrast with 5% (3 of 55) in the cases with clinically N0 ipsilateral necks (p < .05). Based on the clinical staging of the tumor, 8% (3 of 37) of the cases showed lymph node metastases in T2 tumors, 25% (2 of 8) in T3, and 18% (2 of 11) in T4. None of the T1 tumors (10 cases) had pathologically positive lymph nodes. The rate of contralateral occult neck metastasis was significantly higher in advanced-stage cases and those crossing the midline, compared with early-stage or unilateral lesions (p < .05). Patients with no evidence of contralateral nodal cancer had significantly improved disease-specific survival over patients with any pathologically positive nodes (5-year disease-specific survival rate was 79% vs. 43%, p < .05). CONCLUSIONS: The risk of contralateral occult neck involvement in the oral cavity squamous cell carcinomas above the T3 classification or those crossing the midline with unilateral metastases was high, and patients who presented with a contralateral metastatic neck had a worse prognosis than those whose disease was staged as N0. Therefore, we advocate an elective contralateral neck treatment with surgery or radiotherapy in patients with oral cavity squamous cell carcinoma with ipsilateral node metastases or tumors, or both, whose disease is greater than T3 or crossing the midline. PMID- 16721744 TI - Mouse models for human head and neck squamous cell carcinomas. AB - Mouse models of human cancer play an important role in understanding the mechanisms of carcinogenesis and have accelerated the search for finding new molecular targets for cancer therapy. However, genetically engineered mouse models for head and neck squamous cell carcinoma (HNSCC) have only recently overcome major technical obstacles and begun to be explored. Here we review the current progress in the development of mouse models for human HNSCC, with emphasis on conditional transgenic and knockout mouse models. These new models faithfully recapitulate human HNSCC at both the pathologic and molecular levels. These animal models will not only be useful to define the roles of specific genes in HNSCC development and progression but will also provide a unique tool for developing and testing new therapeutic approaches. PMID- 16721745 TI - Primary and salvage (hypo)pharyngectomy: Analysis and outcome. AB - BACKGROUND: Surgery for squamous cell carcinoma (SCC) arising or extending to the hypopharynx is generally reserved for advanced disease or salvage. The prognosis of patients requiring pharyngectomy is poor, and the perioperative morbidity is significant. The aim of the present study is to describe the disease-related and treatment-related outcomes of patients undergoing primary and salvage pharyngectomy for cancer of the hypopharynx and larynx over a 10-year period from a single institution. METHODS: We retrospectively reviewed 138 partial and circumferential pharyngectomies performed at a tertiary referral center between 1992 and 2002. There were 31 females and 107 males. The median age was 62 years (range, 27-81 years), and mean follow-up was 3.6 years. Salvage pharyngectomy for radiation failure was performed in 72 patients (52%), and in 66 patients (48%) pharyngectomy was performed as the primary treatment. RESULTS: The 5-year overall survival rate after salvage pharyngectomy was 31% and after primary pharyngectomy was 38%. The 5-year disease-specific survival (DSS) for salvage was 40% and after primary surgery was 45%. The 5-year local and regional control rates for salvage pharyngectomy were 71% and 70%, respectively, and for primary pharyngectomy were 79% and 67%, respectively. The perioperative mortality rate was 3.6%, and the combined morbidity rate was 70%. Postoperative hypocalcemia developed in 44% of patients, a pharyngocutaneous fistula developed in 31% of patients, and the long term stricture rate was 15%. Variables adversely affecting DSS on univariate analysis were nodal metastases (p = .044), extracapsular spread (ECS) (p = .006), poorly differentiated tumors (p = .015), lymphovascular invasion (p = .042), and positive tumor margins (p = .026). ECS (p = .023) was the only independent prognostic variable on multivariable analysis; however, there was a trend toward significance for nodal metastases (p = .064) and tumor differentiation (p = .079). CONCLUSION: This study demonstrates that both salvage pharyngectomy and primary surgery for advanced disease are viable options with high locoregional control. However, this represents a high-risk group in terms of both operative morbidity and survival. Patients with nodal metastases, ECS, and poorly differentiated tumors are likely to succumb to their disease and should be selected for adjuvant therapy when possible. PMID- 16721746 TI - Primary Ewing sarcoma of the petrous temporal bone: an exceptional cause of facial palsy and deafness in a nursling. AB - BACKGROUND: Primary Ewing sarcoma affecting the skull base in general and the petrous bone in particular is extremely rare with only 4 reports of Ewing sarcoma arising in the petrous temporal bone in the international medical literature. METHODS: The authors report for the first time a case of a primary Ewing sarcoma of the petrous temporal bone in a 5-month-old nursling, which became apparent with a complete peripheral facial palsy and ipsilateral surdity. RESULTS: The neoformation was treated by systemic chemotherapy and radiation of the tumor region. The diagnostic steps, therapy, and development of the child are described in detail; the literature concerning Ewing sarcoma originating from the skull in general and from the petrous temporal bone in particular is reviewed. CONCLUSIONS: The highlights of this case are an extremely uncommon location, an unusual age of presentation, as well as a unique set of symptoms. PMID- 16721747 TI - Microsatellite instability as an indicator of malignant progression in laryngeal premalignancy. AB - BACKGROUND: Microsatellite instability (MSI) is considered a novel marker of genetic instability, and preliminary studies have shown that it may provide useful information in assessing the risk of malignant progression in preinvasive lesions. METHODS: We analyzed MSI in serial biopsy specimens from 10 patients with preinvasive laryngeal lesions and corresponding metachronous laryngeal cancers compared with biopsy specimens of similar lesions without malignant transformation from 20 subjects in a match-paired analysis. MSI was determined by assessing the status of 14 microsatellite markers (chromosome loci: 2p16, 3q21 24, 4q12, 9p21, 13q14, 16q22.1, 17p12 and 21q21) in DNA biopsy specimens. RESULTS: MSI(+) (aberration at two or more loci) was detected in seven of 10 patients with premalignant lesions progressed to carcinoma, whereas only four of the 20 biopsy specimens from control subjects showed an unstable phenotype (p < .01). Interestingly, preinvasive laryngeal lesions with MSI at hMSH2/hMSH6 loci frequently had instability at one or more additional loci and were considered as MSI(+) (overall in eight of 12 cases: six premalignant lesions progressed to cancer and one without progression of the original laryngeal lesion, p < .01). The immunohistochemical analysis of the hMSH2 protein expression in our series, however, did not suggest its involvement in laryngeal carcinogenesis. CONCLUSIONS: Our study indicates that MSI is more common in preneoplastic laryngeal lesions progressing to cancer, thus suggesting that microsatellite status assessment may be useful in determining the risk of malignant progression in patients with preinvasive laryngeal lesions for whom chemopreventive and multiple endoscopic protocols can be attempted. PMID- 16721748 TI - Induction of the transferrin receptor gene by benzo[a]pyrene in breast cancer MCF 7 cells: potential as a biomarker of PAH exposure. AB - Polycyclic aromatic hydrocarbons (PAHs) are widespread environmental DNA-damaging agents regarded as risk factors for human disease, including lung and breast cancer. The biotransformation of PAHs to carcinogenic metabolites is mediated by the aromatic hydrocarbon receptor (AhR), which activates transcription at xenobiotic responsive elements (XREs = 5'-GCGTG-3') found in the promoter regions of genes encoding for detoxifying enzymes, including CYP1A1 and CYP1B1. In this study, we wished to identify novel biomarkers that may be useful in monitoring critical carcinogenic events of the breast induced by PAHs. Using a GeneMAP CancerArray, we analyzed in breast cancer MCF-7 cells the temporal effects of the AhR agonist benzo[a]pyrene (B[a]P), which is a prototype PAH and known environmental carcinogen. Genes upregulated at least threefold by B[a]P and containing potential XREs within their promoter regions included CYP1A1, CYP1B1, paired box gene 3 (PAX3), cortactin (CTTN/EMS1), beta-2-microglobulin (B2M), and transferrin receptor (TfR). The stimulatory effects of B[a]P on expression of these genes were abrogated by cotreatment with the AhR antagonist flavonoid, alpha-napthoflavone (ANF). The TfR gene was selected for further analysis as its promoter region contains two potential XREs and its expression has been shown to be increased in breast cancer cells. Accumulation of TfR mRNA in B[a]P-treated cells was confirmed by quantitative real time PCR. Transient transfection studies indicated that the transcriptional activity of the TfR promoter was stimulated by B[a]P, whereas ANF counteracted this induction. These results indicate that the TfR gene may be a potential biomarker of PAH exposure. PMID- 16721749 TI - Different p53 mutation patterns in colorectal tumors from smokers and nonsmokers. AB - Epidemiological studies consistently find associations between colorectal cancer and cigarette smoking; however, there are little molecular data supporting the association. To examine the relationship between cigarette smoking and colorectal cancer, we compared p53 mutation patterns in colorectal tumors from smokers and nonsmokers. In this study, 153 tumor tissues from colorectal cancer patients, including 63 smokers and 90 nonsmokers, were examined for p53 mutation and p53 protein expression by direct sequencing and immunohistochemistry (IHC), respectively. p53 mutations were detected in 77 of 153 (50.3%) colorectal tumors, and no difference was observed in the p53 mutation frequencies in tumors from smokers and nonsmokers (33 of 63, 50.8% for smokers vs. 44 of 90, 48.9% for nonsmokers, P = 0.743). IHC showed that p53-immunoreactive tumors were positively correlated with p53-mutated tumors (P < 0.0001). G:C-->A:T transition and G:C- >T:A transversion were the predominant types of mutations detected in the tumor p53 genes. G:C-->A:T mutation was relatively more common in nonsmokers than in smokers (93.5% for nonsmokers vs. 77.3% for smokers), although this difference was not significant. The frequency of deletion mutation in smoker tumors, however, was significantly higher than that in nonsmoker tumors (7 of 33, 21.2% for smokers vs. 1 of 44, 2.3% for nonsmokers, P = 0.01). Although there were only a few cases of p53 deletion mutation in this study, the observation of a higher frequency of p53 deletion mutation in smoker tumors supports the association between cigarette smoking and the development of colorectal cancer. PMID- 16721750 TI - Expectation and the placebo effect in Parkinson's disease patients with subthalamic nucleus deep brain stimulation. AB - To determine whether the degree to which a patient with Parkinson's disease expects therapeutic benefit from subthalamic nucleus-deep brain stimulation (STN DBS) influences the magnitude of his or her improved motor response, 10 patients with idiopathic Parkinson's and bilateral STN-DBS were tested after a 12-hour period off medication and stimulation. Four consecutive UPDRS III scores were performed in the following conditions: (a) stimulation OFF, patient aware; (b) stimulation OFF, patient blind; (c) stimulation ON, patient aware; and (d) stimulation ON, patient blind. Statistical significance (P = 0.0001) was observed when comparing main effect ON versus OFF (mean ON: 32.55; mean OFF: 49.15). When the stimulation was OFF, patients aware of this condition had higher UPDRS motor scores than when they were blinded (mean: 50.7 vs. 47.6). With the stimulation ON, UPDRS motor scores were lower when the patients were aware of the stimulation compared with when they were blinded (mean: 30.6 vs. 34.5). The interaction between these levels was significant (P = 0.049). This variation was important for bradykinesia and was not significant for tremor and rigidity. The authors conclude that the information about the condition of the stimulation enhanced the final clinical effect in opposite directions. The results presented support the role of expectation and placebo effects in STN-DBS in Parkinson's disease patients. PMID- 16721751 TI - Two cases of ischemia associated with subthalamic nucleus stimulator implantation for advanced Parkinson's disease. AB - Deep brain stimulation is generally a safe and effective method of alleviating motor impairment in advanced-stage Parkinson's disease patients. However, adverse events of surgery have been noted, such as hemorrhage, infection, seizures, and device failure. In this report, we describe 2 cases of the unusual adverse event of ischemia associated with subthalamic nucleus stimulator implantation. We present the intraoperative neurological symptoms, microelectrode recording data, imaging findings, and other correlated events. In the first case, the clinical effects of ischemia were evident intraoperatively and coincided with silence during microelectrode recording from the ischemic region. In the second case, the timing of the ischemic event could not be determined precisely but also was associated with a difficult mapping. Subcortical ischemia may be an underrecognized event that confounds neurophysiological mapping of deep brain structures and affects clinical outcomes. PMID- 16721752 TI - Natural linewidth chemical shift imaging (NL-CSI). AB - The discrete Fourier transform (FT) is a conventional method for spatial reconstruction of chemical shifting imaging (CSI) data. Due to point spread function (PSF) effects, FT reconstruction leads to intervoxel signal leakage (Gibbs ringing). Spectral localization by imaging (SLIM) reconstruction was previously proposed to overcome this intervoxel signal contamination. However, the existence of magnetic field inhomogeneities creates an additional source of intervoxel signal leakage. It is demonstrated herein that even small field inhomogeneities substantially amplify intervoxel signal leakage in both FT and SLIM reconstruction approaches. A new CSI data acquisition strategy and reconstruction algorithm (natural linewidth (NL) CSI) is presented that eliminates effects of magnetic field inhomogeneity-induced intervoxel signal leakage and intravoxel phase dispersion on acquired data. The approach is based on acquired CSI data, high-resolution images, and magnetic field maps. The data are reconstructed based on the imaged object structure (as in the SLIM approach) and a reconstruction matrix that takes into account the inhomogeneous field distribution inside anatomically homogeneous compartments. Phantom and in vivo results show that the new method allows field inhomogeneity effects from the acquired MR signal to be removed so that the signal decay is determined only by the "natural" R2 relaxation rate constant (hence the term "natural linewidth" CSI). PMID- 16721753 TI - Familial essential tremor with apparent autosomal dominant inheritance: should we also consider other inheritance modes? AB - A positive family history is present in many patients with essential tremor (ET), but twin studies and segregation analysis have suggested that ET is not entirely a genetic disorder. Two genetic loci have been identified in autosomal dominant (AD) ET and polymorphisms in the DRD3 and HS1-BP3 genes have been proposed as the possible susceptibility factors for ET. There is also evidence for further genetic heterogeneity. We evaluated 4 unrelated large kindreds with ET with an apparent AD mode of transmission. Each kindred spanned at least 3 generations and contained at least 13 living affected subjects who met criteria for definitive ET. None of the pedigrees had evidence for inheritance of ET from both parents. Known genetic ET loci were excluded in these families. We detected a preferential transmission of ET in every kindred and the proportion of affected offspring varied from 75% to 90% (P < 0.05) in the generations with complete ascertainment. Our data indicate that non-Mendelian preferential transmission of an affected allele is a feature in many ET kindreds with multiple affected members and an apparent AD mode of inheritance. ET may have a complex etiology. Additional genetic models need to be considered, including an interaction of susceptibility genes and environmental risk factors. PMID- 16721754 TI - The effects of surface lactone hydrolysis and temperature on the specific and nonspecific interactions between phenobarbital and activated carbon surfaces. AB - The effect of hydrolyzing lactone functional groups on the surfaces of different activated carbons upon the specific and nonspecific interactions between phenobarbital and activated carbon surfaces was studied. The effect of temperature on both specific and nonspecific interactions was also studied. The increase in OH groups on the surfaces of activated carbons, as a result of hydrolyzing surface lactone groups, caused an increase in the specific adsorption capacity (K(2)) for phenobarbital without having a significant effect on the hydrophobic bonding capacity (K(HB)). Increasing the temperature at which the adsorption experiment was carried out, on the other hand, resulted in a decrease in K(HB) without having a significant effect on K(2). The decrease in K(HB) per unit temperature increase was the same regardless of the activated carbon. These results are in very good agreement with the modified-Langmuir-like equation (M LLE). PMID- 16721755 TI - Bioadhesive monolayer film for the in vitro transdermal delivery of sumatriptan. AB - The work presented here aims to develop a bioadhesive monolayer film containing sumatriptan as adjuvant for the treatment of headache pain in a severe migraine attack. Permeation experiments were performed from the films prepared and from the respective solution, to evaluate the relevant permeation parameters. The effect of the penetration enhancers Transcutol, 2-pyrrolidone, and polyethylene glycol 600 was evaluated. The results obtained show that Transcutol and 2 pyrrolidone decreased sumatriptan permeation from solution, whereas a modest increase was produced by polyethylene glycol 600. The enhancers produced the same effects when they were included in the film. Compared to solution, the film showed a higher sumatriptan flux in the early times of the experiment. When the film was applied in occlusive conditions the profiles were much higher, indicating the importance of patch drying. Concerning skin retention, the bioadhesive film produced a reduction of the amount of sumatriptan remaining in the skin, but this can be advantageous in the control of drug input, since it reduces the reservoir effect in the skin and allows for an immediate interruption of drug input when the patch is removed. PMID- 16721756 TI - Striatal dopa and glucose metabolism in PD patients with freezing of gait. AB - In Parkinson's disease (PD), freezing suggests sudden and transient blocks of motor behavior during initiating or continuous repetitive movements. Its underlying pathophysiology remains unclear. The objective of this study is to compare striatal dopamine metabolism and cerebral glucose metabolism between PD patients with and without freezing of gait (FOG). A total of 10 PD patients with FOG at off and 7 PD patients without FOG underwent brain positron emission tomography with (18)[F]-6-fluoro-levodopa (FDOPA) and (18)[F]-fluordesoxyglucose (FDG). Striatum decarboxylase activity was expressed by metabolic influx constants of the striatum related to the occipital lobe (Kocc). FDG uptake in caudate and putamen was normalized to global FDG uptake. Region of interest (ROI) analysis of striatal regions was used, as well as voxel-based analysis by statistical parametric mapping (SPM). ROI analysis did not reveal differences in striatal FDOPA and FDG uptake between the groups. SPM showed lower putaminal FDOPA uptake (P = 0.05 uncorrected) with increased FDG uptake (P = 0.01 uncorrected) in freezing PD, whereas caudate uptake of the two tracers was reduced. Freezing-related cortical FDG decrease was found in (right) parietal regions. In conclusion, in freezing PD, caudate uptake of FDG and FDOPA was reduced, whereas putamen FDOPA decrease was associated with FDG increase. Right hemisphere circuitry seemed to be more affected in freezing patients. PMID- 16721757 TI - Selectively crosslinked hyaluronic acid hydrogels for sustained release formulation of erythropoietin. AB - A novel sustained release formulation of erythropoietin (EPO) was developed using hyaluronic acid (HA) hydrogels. For the preparation of HA hydrogels, adipic acid dihydrazide grafted HA (HA-ADH) was synthesized and analyzed with (1)H NMR. The degree of HA-ADH modification was about 69%. EPO was in situ encapsulated into HA ADH hydrogels through a selective cross-linking reaction of bis(sulfosuccinimidyl) suberate (BS(3)) to hydrazide group (pK(a) = 3.0) of HA ADH rather than to amine group (pK(a) > 9) of EPO. The denaturation of EPO during HA-ADH hydrogel synthesis was drastically reduced with decreasing pH from 7.4 to 4.8. The specific reactivity of BS(3) to hydrazide at pH = 4.8 might be due to its low pK(a) compared with that of amine. In vitro release of EPO in phosphate buffered saline at 37 degrees C showed that EPO was released rapidly for 2 days and then slowly up to 4 days from HA-ADH hydrogels. When the hydrogels were dried at 37 degrees C for a day, however, longer release of EPO up to 3 weeks could be demonstrated. According to in vivo release test of EPO from HA-ADH hydrogels in SD rats, elevated EPO concentration higher than 0.1 ng/mL could be maintained from 7 days up to 18 days depending on the preparation methods of HA-ADH hydrogels. There was no adverse effect during and after HA-ADH hydrogel implantation. PMID- 16721758 TI - Collagen I-coated titanium surfaces: mesenchymal cell adhesion and in vivo evaluation in trabecular bone implants. AB - The goal of the study was the evaluation of the effect of modification of titanium implants by acrylic acid surface grafting-collagen I coupling. Tests were performed on titanium samples treated by galvanostatic anodization to create a porous surface topography. Surface characterization by X-ray photoelectron spectroscopy (XPS) and scanning electron microscopy (SEM) confirms the biochemical modification of the surface and shows a surface topography characterized by pores mostly below 1 mum diameter. In vitro evaluation involving human mesenchymal cells shows enhanced cell growth on collagen coated surfaces as compared to titanium ones. Four weeks in vivo evaluation of implants in rabbit femur trabecular bone shows improvements of bone-to-implant contact, while improvement of bone ingrowth is slightly not significant (p = 0.056), when compared to the control. Overall, these data indicate that integration in trabecular, or cancellous, bone can be enhanced by the surface collagen layer, confirming previous findings obtained by modification of machined surfaces by the same approach in cortical bone implants. PMID- 16721759 TI - Differential expression of voltage-activated calcium currents in zebrafish retinal ganglion cells. AB - We report a study on the characterization of voltage-activated calcium currents (I(Ca)) in retinal ganglion cells (RGCs) and the topographic distribution of RGCs that express different types of I(Ca) in zebrafish retinas. In acutely isolated zebrafish RGCs, both high-voltage-activated (HVA; peak activation potential +7.4 +/- 1.1 mV) and low-voltage-activated (LVA; peak activation potential -33.0 +/- 1.2 mV) I(Ca) were recorded. HVA I(Ca) were recorded in all of the tested RGCs, whereas LVA I(Ca) were recorded in approximately one-third of the tested cells. In RGCs that expressed both HVA and LVA I(Ca), the two currents were readily separated by depolarizing the cell membrane to different voltages from different holding potentials. Among RGCs that expressed LVA I(Ca), some cells expressed large LVA I(Ca) (up to 130 pA), whereas others expressed small LVA I(Ca) (approximately 20 pA). RGCs that expressed large and small LVA I(Ca) were designated as class I and class II cells, respectively, and RGCs that expressed only HVA I(Ca) were designated as class III cells. The topographic distribution of cell classes was similar in various areas of the retina. In the nasal-ventral retina, for example, class III cells outnumbered class I and class II cells by 10.8- and 2.6-fold, respectively. In the temporal and dorsal retinas, the density of class III cells slightly decreased, whereas the density of class I and class II cells increased. The differential expression of I(Ca) in RGCs may correlate with the development and function of the retina. PMID- 16721760 TI - Absence of synaptophysin near cortical neurons containing oligomer Abeta in Alzheimer's disease brain. AB - Soluble amyloid beta protein (Abeta) oligomers have been considered recently to be responsible for the cognitive dysfunction that sets in prior to senile plaque formation in the Alzheimer's disease (AD) brain. By using the newly prepared antibody against oligomer Abeta, rather than fibrillar or monomer Abeta, we observed that oligomer Abeta in AD brains was localized as clusters ofdot likeimmunostains in the neurons in a manner different from that in senile plaques. The relationship of oligomer Abeta with synaptophysin, a synaptic molecular marker, was examined because oligomer Abeta is widely believed to be related to synaptic failure. We observed that immunostainings for synaptophysin were absent near neurons bearing clusters of oligomer Abeta. The present study provides morphological evidence to support the idea that accumulated oligomer Abeta, but not fibrillar Abeta, is closely associated with synaptic failure, which is the major cause of cognitive dysfunction. PMID- 16721761 TI - Glutathione peroxidase 1 and glutathione are required to protect mouse astrocytes from iron-mediated hydrogen peroxide toxicity. AB - The enzyme glutathione peroxidase 1 (GPx1) is involved in the cellular detoxification of peroxides. To test for the consequences of GPx deficiency in astrocytes, astrocyte-rich primary cultures from wild-type and GPx1-deficient [GPx1(-/-)] mice were exposed to H(2)O(2). In GPx1(-/-) astrocytes, the clearance rate of H(2)O(2) was slower than in wild-type cells. In contrast to GPx1 deficient astrocytes, wild-type cells exhibited, within 2 min of H(2)O(2) application, a rapid and transient accumulation of cellular glutathione disulfide that amounted to 60% of total glutathione. The peroxide treatment did not affect the viability of wild-type astrocytes, whereas 45% of the GPx1(-/-) cells died within 8 hr. However, the viability of both types of astrocytes was strongly compromised by lowering cellular glutathione content before peroxide application. In contrast, inactivation of catalase caused substantial cell death only in GPx1( /-) cells but not in wild-type astrocytes. The cell death observed was prevented by the iron chelators deferoxamine, 1,10-phenathroline, or 2,2'-dipyridyl, whereas preincubation with ferric ammonium citrate increased the toxicity of peroxide treatments. These results demonstrate that GPx1 contributes to the rapid clearance of H(2)O(2) by mouse astrocytes and that both GPx1 and a high concentration of glutathione are required to protect these cells from iron dependent peroxide damage. PMID- 16721762 TI - Heavy chain of cytoplasmic dynein is a major component of the postsynaptic density fraction. AB - A protein with an apparent molecular size of 490 kDa was found in the postsynaptic density (PSD) fraction isolated from porcine cerebral cortices and rat forebrains, and this 490 kDa protein accounted for approximately 3% of the total protein of these samples. Matrix-assisted laser desorption ionization-time of flight mass spectrometric and Western blotting analyses consistently indicated that this 490 kDa protein consisted primarily of the heavy chain of cytoplasmic dynein (cDHC). Immunocytochemical analyses showed that cDHC was found in 92% and 89% of the phalloidin-positive protrusions that were themselves associated with discrete clusters of synaptophysin, a presynaptic terminal marker, and PSD-95, a postsynaptic marker, on neuronal processes, respectively. Quantitative Western blotting analyses of various subcellular fractions isolated from porcine cerebral cortices and rat forebrains further showed that not only the heavy but also the intermediate chains of dynein are enriched in the PSD fraction. Cytoplasmic dynein is a microtubule-associated motor protein complex that drives the movement of various cargos toward the minus ends of microtubules and plays many other diverse functions in the cell. Our results that cDHC is a major component of the PSD fraction, that both dynein heavy and intermediate chains are enriched in the PSD fraction and that cDHC is present in dendritic spines raise the possibilities that cytoplasmic dynein may play structural and functional roles in the postsynaptic terminal. PMID- 16721763 TI - Successful treatment with thalidomide in a liver transplant recipient with giant esophageal ulcers. PMID- 16721764 TI - Fulminant hepatic failure associated with the use of black cohosh: a case report. AB - With data from the Women's Health Initiative indicating that estrogen plus progesterone are associated with an increased risk of cardiovascular events, many patients and practitioners are looking for alternative therapies to manage menopausal symptoms. One alternative is black cohosh, an herbal product used primarily to treat these symptoms. In recent years there have been several case reports associating this substance with hepatitis and fulminant hepatic failure. We present a case of a woman who developed hepatic failure requiring liver transplantation from the use of this herb. PMID- 16721765 TI - Five-year follow-up of a hepatitis B virus-positive recipient of hepatitis B surface antigen-positive living donor liver graft. AB - The shortage of cadaveric donor organs has led to the use of living donors and marginal cadaveric donors. To date, there have been only 2 reports on the use of hepatitis B surface antigen (HBsAg)-positive liver grafts. Here we describe the 5 yr posttransplantation sequence of a hepatitis B virus (HBV)-positive recipient who received an HBsAg-positive living donor liver graft. A 43-yr-old HBV-positive patient with hepatorenal syndrome received a living donor liver graft in October 2000 from a 27-yr-old HBsAg-positive carrier with no clinical evidence of HBV infection other than the serologic markers. The recipient recovered slowly after liver transplantation (LT). Recipient serum HBsAg was continuously positive despite anti-HBV therapy with high-dose hepatitis B immunoglobulin (HBIG) and lamivudine. The patient was also treated with famciclovir and interferon; to date, a final regimen of lamivudine and adefovir has kept liver function stable for 20 months. The recipient has lived for 64 months after transplantation. The donor has not revealed any clinical evidence of active hepatitis during follow up. In conclusion, our result implicates that a recipient of liver graft from an HBsAg-positive carrier may survive for a long period following antiviral therapy with lamivudine and adefovir. Considering this living donor case and previously reported cases, the use of an HBsAg-positive cadaveric liver graft may deserve attention when no other donor is available. PMID- 16721766 TI - Study of tryptophan metabolism via serotonin in cerebrospinal fluid of patients with noncommunicating hydrocephalus using a new endoscopic technique. AB - By a recent minimally invasive neuroendoscopic technique, the cerebral ventricles have been reached in a quick, reliable, and harmless way, making possible the study of cerebrospinal fluid (CSF) of the lateral ventricles and, above all, the CSF adjacent to the walls of the third ventricle. Tryptophan, 5 hydroxytryptophan, serotonin (5-HT), and 5-hydroxyindoleacetic acid (5-HIAA) were measured in CSF by HPLC equipment. Twenty-six patients affected with noncommunicating hydrocephalus were enrolled in the study and, as controls, 28 subjects not suffering from any neurological disease. The concentrations of tryptophan were higher in right ventricular CSF than in lumbar CSF (P < 0.01). 5 HT was detectable in the CSF of the right ventricle of hydrocephalic patients. 5 HIAA was higher in right ventricular CSF than in cisternal and lumbar CSF (P < 0.01), both in controls and in hydrocephalic patients. However, there was a higher concentration of 5-HIAA in right ventricular (P < 0.05) and cisternal (P < 0.01) CSF in hydrocephalic patients in comparison with controls. In the CSF samples withdrawn during neuroendoscopy, 5-HT presented the highest concentrations in the pineal recess. The highest amounts of 5-HIAA were found in the choroid plexus, third and right ventricles, pituitary recess, and aqueduct, and the lowest in pineal recess, subarachnoid space, infundibulum, and interpeduncolar cistern. These results provide new insight into the fate of tryptophan and its metabolites via serotonin in the CSF and suggest the feasibility of the new neuroendoscopic technique for brain metabolic studies. PMID- 16721767 TI - Cellular composition of long-term human spinal cord- and forebrain-derived neurosphere cultures. AB - In vitro expanded neural precursor cells (NPCs) may provide a stable source for cell therapy. In search of the optimal cell source for spinal cord repair, we investigated influences of gestational age, regional heterogeneity, and long-term in vitro propagation. The cellular content of neurosphere cultures prior to and after in vitro differentiation was studied by immunocytochemistry and flow cytometry. Human forebrain and spinal cord NPCs deriving from first-trimester tissue were cultured as neurospheres in the presence of epidermal growth factor, basic fibroblast growth factor, and ciliary neurotrophic factor. Proteins characteristic for embryonic stem cells, i.e., Tra-1-60, Tra-1-81, and SSEA-4, were present in approximately 0.5% of the cells in donor tissues and neurospheres. The proportions of nestin- and proliferating cell nuclear antigen immunoreactive (IR) cells were also maintained, whereas the CD133-IR population increased in vitro. Glial fibrillary acidic protein-IR cells increased in number, and in contrast the fraction of beta-tubulin III-IR cells decreased, at and beyond passage 5 in spinal cord but not forebrain cultures. However, dissociated and in vitro-differentiated forebrain- and spinal cord-derived neurospheres generated similar proportions of neurons, astrocytes, and oligodendrocytes. Gestational age of the donor tissue, which ranged from 4.5 to 12 weeks for forebrain and from 4.5 to 9.5 weeks for spinal cord, did not affect the proportion of cells with different phenotypes in culture. Thus, cellular composition of human neurosphere cultures differs as a result of long-term in vitro propagation and regional heterogeneity of source tissue, despite expansion under equal culture conditions. This could in turn imply that human spinal cord and forebrain NPCs present different repair potentials in in vivo settings. PMID- 16721768 TI - Living donor liver transplantation for hepatocellular carcinoma exceeding conventional criteria: questions, answers and demands for a common language. PMID- 16721772 TI - Safe liver harvesting from living donors. PMID- 16721773 TI - Intraoperative blood loss is a risk factor for complications in donors after living donor hepatectomy. AB - Complications in a donor are a distressing but inevitable occurrence, since graft procurement is a major undertaking. Although the technique for procurement has some similarities to hepatic resection, a donor is very unlike a patient with malignancy. The risk factors identified in these patients cannot be extrapolated to donors. Donor hepatectomy carried out from June 1995 to March 2005 in Chang Gung Memorial Hospital, Kaohsiung Medical Center was reviewed with the aim of identifying risk factors for complications. There were 204 living donor liver transplants, with 205 donor hepatectomies, as 1 living donor liver transplantation was a dual graft. Ten donors (4.88%) suffered complications. There was no difference in terms of age, gender, body weight, operation, and parenchymal time between those who had complications and those who did not. There was also no difference in liver function tests between the 2 groups of donors, but the total bilirubin was significantly higher in donors with complications. The graft weight and remnant liver volume were also similar. The proportion of donors with fatty liver was the same between the 2 groups. The mean blood loss in donors with complications was 170 +/- 79 mL, and that for donors without complications was 95 +/- 77 mL. There was a statistically significant greater blood loss in donors with complications (P < 0.05). The number of segments removed in donors with complications was also higher compared to donors without complications (P < 0.03). Using multivariate analysis, intraoperative blood loss and the number of segments removed were found to be independent risk factors for donor complications. Intraoperative blood loss during graft procurement must be kept low to minimize complications in donors. PMID- 16721774 TI - Successful ABO-incompatible pediatric liver transplantation utilizing standard immunosuppression with selective postoperative plasmapheresis. AB - Transplanting blood group A, B, or O (ABO)-incompatible (ABO-I) liver grafts has resulted in lower patient and graft survival with an increased incidence of vascular and biliary complications and rejection. We report that, without modification of our standard immunosuppression protocol, crossing blood groups is an acceptable option for children requiring liver transplantation. In our study, ABO-I liver grafts -- regardless of recipient age -- have comparable long-term survival (mean follow-up of 3.25 yr) with ABO-compatible grafts without any difference in rejection, vascular or biliary complications. From January 1, 1999 to October 1, 2005, we studied 138 liver transplants in 121 children: 16 (13.2%) received an ABO incompatible liver allograft. One-year actuarial patient survival for ABO-matched grafts vs. ABO-I grafts was 93.0% and 100%, respectively, whereas graft survival was 83.4% and 92.3%. Additionally, 6 of 16 (37.5%) ABO-I transplanted children had 8 rejection episodes, whereas 47 patients (44.8%) had 121 rejection episodes in the ABO-compatible group. There were no vascular complications and 2 biliary strictures in the ABO-I group. Plasmapheresis was not used for pretransplantation desensitization and was only required in 1 posttransplantation recipient. No child was splenectomized. Six of the 16 children were older than 13 yr of age, suggesting the possibility of successfully expanding this technique to an older population. In conclusion, our outcomes may support the concept of using ABO-I grafts in a more elective setting associated with split and living donor liver transplants. PMID- 16721775 TI - Living donor liver transplantation for Budd-Chiari syndrome using cryopreserved vena cava graft in retrohepatic vena cava reconstruction. PMID- 16721776 TI - Consensus conference: Indications for Liver Transplantation, January 19 and 20, 2005, Lyon-Palais Des Congres: text of recommendations (long version). PMID- 16721777 TI - Biliary strictures in living donor liver transplantation: incidence, management, and technical evolution. AB - Biliary complications, biliary strictures (BS) in particular, continue to be a significant cause of morbidity after LDLT despite technical refinement. In this study, we assessed the incidence of BS and their management in living donor liver transplantation (LDLT) with special reference to the type of biliary reconstruction. A total of 182 LDLTs performed at our institution for either adult (n = 157) or pediatric (n = 25) patients were included in the study. The duct-to-duct (DD) biliary reconstruction was performed for 106 cases, while the conventional Roux-en-Y hepaticojejunostomy (HJ) was utilized for the remaining 76 cases. Overall, BS developed in 46/182 (25.3%) of the cases (DD, 26.4%; HJ, 25.0%). The 1- and 3-year cumulative incidences of BS were 22.9% and 31.9%, respectively, in the DD group, and 15.2% and 29.1%, respectively, in the HJ group (P= not significant). The left-lobe LDLT was more prone to develop BS. Continuous anastomosis tended to be associated with the high incidence of BS in the DD group. The incidence of anastomotic leak was significantly lower in the DD group. Intervention via either pre-cutaneous or endoscopic approach was successful in the majority of cases, although recurrence could occur in some patients. In conclusion, BS was not associated with the type of reconstruction in LDLT. The primary radiological or endoscopic interventions were satisfactory treatments of choice. Technical refinement is an important factor to reduce the incidence of BS. PMID- 16721778 TI - Question of using valganciclovir for cytomegalovirus (CMV) infection prophylaxis in post-liver transplant recipients. PMID- 16721779 TI - Determining the nature and impact of fatigue after liver transplantation. PMID- 16721780 TI - Lessons learned from 1,000 living donor liver transplantations in a single center: how to make living donations safe. AB - Serious complications have occurred in a considerable proportion of living donors of liver transplants, but data from a single high-volume center has rarely been available. We analyzed the medical records of donors and recipients of the first 1,000 living donor liver transplants, performed at Asan Medical Center from December 1994 to June 2005, with a focus on donor safety. There were 107 pediatric and 893 adult transplants. The most common diagnoses were biliary atresia in pediatric recipients (63%) and hepatitis B-associated liver cirrhosis (80%) in adult recipients. Right lobe donors were strictly selected based on liver resection rate and steatosis. From 1,162 living donors, 588 right lobes, 6 extended right lobes, 7 right posterior segments, 464 left lobes, and 107 left lateral segments were obtained. Of these, 837 grafts were implanted singly, whereas 325, along with 1 cadaveric split graft, were implanted as dual grafts into 163 recipients. The 5-yr survival rates were 84.8% in pediatric recipients and 83.2% in adult recipients. There was no donor mortality, but 3.2% of donors experienced major complications. Until the end of 2001, the major donor complication rate was 6.7%, with most occurring in right liver donors. Since 2002, liver resection exceeding 65% of whole liver volume were avoided except for young donors with no hepatic steatosis, and the donor complication rate has been reduced to 1.3%. In conclusion, a majority of major living donor complications appear to be avoidable through the strict selection of living donor and graft type, intensive postoperative surveillance, and timely feedback of surgical techniques. Selection of right lobe graft should be very prudently considered if the donor right liver appears to be larger than 65% of the whole liver volume. PMID- 16721781 TI - Saying "Yes" to obese living liver donors: short-term intensive treatment for donors with hepatic steatosis in living-donor liver transplantation. AB - BACKGROUND: The use of steatotic livers is associated with increased primary nonfunction in liver transplantation. To reduce the risk of liver injury, we applied a short-term combination therapy of diet, exercise and drugs for 11 living-donor liver transplantation (LDLT) candidates with steatosis. METHODS: Subjects were treated with a protein-rich (1000 kcal/day) diet, exercise (600 kcal/day), and bezafibrate (400 mg/day) for 2-8 weeks. RESULTS: The treatment significantly improved macrovesicular steatosis (30+/-4% vs. 12+/-2% [mean+/ SEM], P=0.0028). Body weight and BMI were significantly reduced (73.7+/-3.2 kg vs. 66.9+/-2.9 kg, P=0.0033, 26.4+/-0.7 kg/m(2) vs. 24.1+/-0.8 kg/m(2), P=0.0033). The treatment completely normalized liver function tests and lipid metabolism. Seven treated liver grafts (left lobe) were transplanted to the recipients. We compared transplanted graft function and resected liver function of donors using parameters such as peak total bilirubin, prothrombin time at postoperative day 3, and peak alanine aminotransferase between treated liver (n=7) and donor liver without hepatic steatosis (n=37). The transplanted grafts showed good liver functions, and there was no difference between them with respect to functional parameters. The treated donors also showed good liver functions, and no significant differences in functional parameters. CONCLUSIONS: The results of this study indicate that our short-term treatment effectively reduced steatosis and contributed to safer LDLT. Our findings also suggest that even severely steatotic livers can be used for LDLT grafting subsequent to our short-term treatment regimen. PMID- 16721782 TI - Food intake and risk of squamous cell carcinoma of the skin in a community: the Nambour skin cancer cohort study. AB - There is some evidence that dietary factors may modify the risk of squamous cell carcinoma (SCC) of the skin, but the association between food intake and SCC has not been evaluated prospectively. We examined the association between food intake and SCC incidence among 1,056 randomly selected adults living in an Australian sub-tropical community. Measurement-error corrected estimates of intake in 15 food groups were defined from a validated food frequency questionnaire in 1992. Associations with SCC risk were assessed using Poisson and negative binomial regression to the persons affected and tumour counts, respectively, based on incident, histologically confirmed tumours occurring between 1992 and 2002. After multivariable adjustment, none of the food groups was significantly associated with SCC risk. Stratified analysis in participants with a past history of skin cancer showed a decreased risk of SCC tumours for high intakes of green leafy vegetables (RR = 0.45, 95% CI = 0.22-0.91; p for trend = 0.02) and an increased risk for high intake of unmodified dairy products (RR = 2.53, 95% CI: 1.15-5.54; p for trend = 0.03). Food intake was not associated with SCC risk in persons who had no past history of skin cancer. These findings suggest that consumption of green leafy vegetables may help prevent development of subsequent SCCs of the skin among people with previous skin cancer and that consumption of unmodified dairy products, such as whole milk, cheese and yoghurt, may increase SCC risk in susceptible persons. PMID- 16721783 TI - Mannose-binding lectin-2 genetic variation and stomach cancer risk. AB - Deficiency of the mannose-binding lectin (MBL) protein, an antigen-recognition molecule involved in systemic and mucosal innate immunity, is determined by variant alleles in MBL2 gene promoter and exon-1 regions. We conducted a population-based study on 305 stomach cancer cases and 427 controls in Warsaw, Poland to determine whether MBL2 gene variants predispose to stomach cancer. Single nucleotide polymorphisms (SNPs) in MBL2 were determined by TaqMan. The 5 tested MBL2 variants are in complete linkage disequilibrium and comprise 6 different haplotypes. The risk of stomach cancer was increased in subjects carrying the H/H promoter genotype (OR = 1.8, 95%CI 1.1-2.9; p = 0.020) relative to L/L carriers, after adjustment for age, gender, education and smoking. Carrying at least one D exon-1 allele was associated with nonsignificant excess risk (OR = 1.5, 95% CI 0.9-2.4; p = 0.081). In haplotype analysis, the HYD haplotype was associated with increased risk of stomach cancer when compared with HYA, the most common haplotype (OR = 1.9, 95% CI 1.1-3.2; p = 0.021). In diplotype analysis, subjects carrying the YA/D haplotype combination showed the highest risk (OR = 3.0, 95% CI 1.2-7.1; p = 0.015), compared with YA/YA. Further analyses to examine the joint effect of MBL2 and IL-1B polymorphisms, previously shown to predispose to stomach cancer, indicated that the combination of at-risk IL-1B genotypes (CT or TT at location -511) and HYD MBL2 haplotype was associated with a 3.5-fold risk (OR = 3.5, 95% CI 1.6-7.6; p = 0.001). Our findings suggest that the codon 52 D MBL2 variant causing a cysteine > arginine replacement, but not B and C variants producing glycine substitutions, is specifically associated with gastric cancer risk. PMID- 16721784 TI - Melanocortin 1 receptor variants and skin cancer risk. AB - Melanocortin 1 receptor (MC1R) gene variants are associated with red hair and fair skin color. We assessed the associations of common MC1R genotypes with the risks of 3 types of skin cancer simultaneously in a nested case-control study within the Nurses' Health Study (219 melanoma, 286 squamous cell carcinoma (SCC), and 300 basal cell carcinoma (BCC) cases, and 873 controls). We found that the 151Cys, 160Trp and 294His variants were significantly associated with red hair, fair skin color and childhood tanning tendency. The MC1R variants, especially the 151Cys variant, were associated with increased risks of the 3 types of skin cancer, after controlling for hair color, skin color and other skin cancer risk factors. Carriers of the 151Cys variant had an OR of 1.65 (95% CI, 1.04-2.59) for melanoma, 1.67 (1.12-2.49) for SCC and 1.56 (1.03-2.34) for BCC. Women with medium or olive skin color carrying 1 nonred hair color allele and 1 red hair color allele had the highest risk of melanoma. A similar interaction pattern was observed for red hair and carrying at least 1 red hair color allele on melanoma risk. We also observed that the 151Cys variant contributed additional melanoma risk among red-haired women. The information on MC1R status modestly improved the risk prediction; the increase was significant for melanoma and BCC (p, 0.004 and 0.05, respectively). These findings indicated that the effects of the MC1R variants on skin cancer risk were independent from self-reported phenotypic pigmentation. PMID- 16721785 TI - BRAF and KRAS mutations in prostatic adenocarcinoma. AB - Constitutive activation of the kinase cascade involving RAS, RAF, MEK and ERK is common to human cancers, and mutations of KRAS and BRAF are mutually exclusive and serve as alternatives to activate the RAS/RAF/ERK signaling pathway. RAS mutations are known to occur in prostate adenocarcinomas, but little is known about BRAF mutations in these tumors. In the present study, BRAF and KRAS mutations were characterized in 206 prostate adenocarcinomas by enhanced PCR-RFLP and direct sequencing. The identified KRAS and BRAF mutations were then analyzed with respect to preoperative serum PSA levels, Gleason scores and tumor stages. Mutations in codon 600 of BRAF were identified in 21 (10.2%) of 206 prostate adenocarcinomas. KRAS mutations in codons 12 or 13 were found in 15 (7.3%) of 206 prostate adenocarcinomas. However, no tumor specimen contained both BRAF and KRAS mutations. Prostate adenocarcinomas with a BRAF mutation tended to show higher preoperative serum PSA levels, Gleason scores and tumor stages than prostate adenocarcinomas with a KRAS mutation. The results obtained show that BRAF mutations are as uncommon as KRAS mutations in prostate adenocarcinoma. Although BRAF and KRAS are members of the same RAS/ERK signaling pathway, prostate adenocarcinomas with a BRAF mutation showed clinicopathologic features that differed from those of prostate adenocarcinoma with a KRAS mutation. PMID- 16721786 TI - Resistance to apoptosis of HCW-2 cells can be overcome by curcumin- or vincristine-induced mitotic catastrophe. AB - The term mitotic catastrophe has recently become widely used to describe a form of death affecting many cancer cells, which, because of severe DNA or mitotic spindle damage, are not able to bypass mitosis. We show here that cells of the HL 60-derived HCW-2 line highly resistant to apoptosis, upon treatment with curcumin or vincristine, undergo mitotic catastrophe that is finalized by caspase 3 activation and oligonucleosomal DNA degradation. Curcumin is a natural dye, derived from Curcuma longa that has been shown to induce cell death in many cancer cells. Both treatments decrease cell proliferation and cell survival, arrest cells in G2/M phase of cell cycle and induce morphological changes characterized by cell enlargement and micronucleation. "Catastrophic" cells comprise a separate subpopulation with less than 4C DNA, as evidenced by flow and scanning cytometry. This subpopulation is MPM-2 positive. Thymidine block increased the number of cell arrested in the G2/M phase of cell cycle and curcumin effectiveness as an inducer of mitotic catastrophe. Curcumin, but not vincristine, acts on HCW-2 cells by inhibiting the expression of survivin, a modulator of cell division and apoptosis in cancer. Altogether our results show that apoptosis resistance can be overcome by inducing mitotic catastrophe in HCW 2 cells. PMID- 16721787 TI - p53 Arg72Pro polymorphism and risk of colorectal adenoma and cancer. AB - A single nucleotide polymorphism (SNP) at codon 72 of the p53 gene (Arg72Pro) alters the p53 protein structure and affects its activity. We investigated this SNP in relation to colorectal adenoma and cancer among men and women from case control studies nested within the Nurses' Health Study, the Health Professionals Follow-up Study and the Physicians' Health Study. Among 856 colorectal adenoma cases and 1,184 controls, we observed a modest association with p53 Arg72Pro genotype (multivariate odds ratio (OR) = 1.25, 95% confidence interval (CI) = 1.04-1.50 for Arg/Pro and Pro/Pro vs. Arg/Arg). This association did not vary by colorectal site or by sex. Among 442 colorectal cancer cases and 904 controls, we observed no significant overall association between p53 Arg72Pro genotype and colorectal cancer (multivariate OR = 1.14, 95% CI = 0.90-1.45). However, when colorectal site and sex was accounted for, the Pro carrier genotypes compared to Arg/Arg were associated with an increased risk of proximal colon cancers in women (multivariate OR = 2.59, 95% CI = 1.49-4.52) though not with distal colon or rectal cancers, while among men the same genotypes were associated with an increased risk of distal colon cancers (multivariate OR = 2.09, 95% CI = 1.28 3.40) but not proximal colon or rectal cancers. Our results suggest that Arg72Pro may play a role in the early stages of colorectal neoplasia and possibly in progression to invasive disease, depending on site and sex. PMID- 16721788 TI - High incidence of EMMPRIN expression in human tumors. AB - Extracellular matrix metalloproteinase inducer expressed by tumor cells stimulates peritumoral fibroblasts to produce matrix metalloproteinases, thus contributing to tumor invasion and metastasis. To assess its suitability as potential therapeutic target, the overall incidence of EMMPRIN expression in normal and neoplastic tissues was analyzed. EMMPRIN expression was detected immunohistochemically using monoclonal antibodies MEM-M6/1 and HIM6 and tissue microarrays with 2,348 and 608 tissue samples from 129 distinct tumor types and 76 different normal tissues, respectively. Expression and glycosylation state of EMMPRIN in human breast cancer cells were analyzed by Western blot analysis with monoclonal antibodies recognizing distinct carbohydrate structures and biochemical methods. EMMPRIN expression was found in 112 of 129 tumor entities analyzed with malignant tumors being EMMPRIN positive more frequently than benign tumors. A remarkable heterogeneity in EMMPRIN expression between tumor entities was observed. Among others, squamous-cell carcinomas (60-100%), pancreatic (87%), chromophobic kidney (83%), hepatocellular (83%) or medullary breast (83%) adenocarcinomas as well as glioblastoma multiforme (79%) presented with a particular high incidence of EMMPRIN expression. There were a limited number of EMMPRIN-positive normal cell types including proliferatively active and differentiating epithelial cells, germ cells, myocardial cells in the left heart ventricle or vascular endothelial cells of the brain. We could further demonstrate that breast cancer cells expressed EMMPRIN isoforms differing in the presence or absence of Lewis X glycan structures. Our results may assist in defining the suitability of EMMPRIN as therapeutic target and predicting negative side effects. PMID- 16721789 TI - MUC2 expression is regulated by histone H3 modification and DNA methylation in pancreatic cancer. AB - Mucins are highly glycosylated proteins that play important roles in carcinogenesis. In pancreatic neoplasia, MUC2 mucin has been demonstrated as a tumor suppressor and we have reported that MUC2 is a favorable prognostic factor. Regulation of MUC2 gene expression is known to be controlled by DNA methylation, but the role of histone modification for MUC2 gene expression has yet to be clarified. Herein, we provide the first report that the histone H3 modification of the MUC2 promoter region regulates MUC2 gene expression. To investigate the histone modification and DNA methylation of the promoter region of the MUC2 gene, we treated 2 human pancreatic cancer cell lines, PANC1 (MUC2-negative) and BxPC3 (MUC2-positive) with the DNA methyltransferase inhibitor 5-azacytidine (5-aza), the histone deacetylase inhibitor trichostatin A (TSA), and a combination of these agents. The DNA methylation level of PANC1 cells was decreased by all 3 treatments, whereas histone H3-K4/K9 methylation and H3-K9/K27 acetylation in PANC1 cells was changed to the level in BxPC3 cells by treatment with TSA alone and with the 5-aza/TSA combination. The expression level of MUC2 mRNA in PANC1 cells exhibited a definite increase when treated with TSA and 5-aza/TSA, whereas 5-aza alone induced only a slight increase. Our results suggest that histone H3 modification in the 5' flanking region play an important role in MUC2 gene expression, possibly affecting DNA methylation. An understanding of these intimately correlated epigenetic changes may be of importance for predicting the outcome of patients with pancreatic neoplasms. PMID- 16721790 TI - Time-resolved rapid-scan Fourier transform infrared difference spectroscopy on a noncyclic photosystem: rhodopsin photointermediates from Lumi to Meta II. AB - The visual pigment rhodopsin has been extensively studied for the kinetics of its photointermediates by various spectroscopic methods. Unlike such archaeal retinal proteins as bacteriorhodopsin, visual rhodopsin does not thermally recover its dark state after photoexcitation, which precludes repeated excitation of a single sample and thereby complicates time-resolved experiments. Kinetic data on the late rhodopsin photointermediates have so far been available mainly from time resolved ultraviolet (UV)-visible spectroscopy, but not from Fourier transform infrared (FTIR) spectroscopy. The latter has the advantage of being informative of structural changes of both chromophore and protein, but does not allow the highly reproducible, automated sample exchange procedures available to UV-visible spectroscopy. Using rapid-scan FTIR difference spectroscopy, we obtained time resolved data sets that were analyzed by a maximum entropy inverse Laplace transform. Covering the time range from 8 ms to 15 s at temperatures of 0 and -7 degrees C, the transitions from the Lumi to the Meta I and from the Meta I to the Meta II photoproduct states could be resolved. In the transition from Meta I to Meta II, our data reveal a partial deprotonation of the retinal Schiff base preceding the conformational change of the receptor protein to Meta II. The technique and the results are discussed in regard to its advantages as well as its limitations. PMID- 16721791 TI - The interaction between N-n-undecyl-N'-(sodium p-aminobenzenesulfonate) thiourea and serum albumin studied using various spectroscopies and the molecular modeling method. AB - The preparation and characteristics of N-n-undecyl-N'-(sodium-p aminobenzenesulfonate) thiourea (UPT), a new water-soluble reagent with a saturated fatty hydrocarbon group, were described. The interactions of UPT with bovine serum albumin (BSA) and human serum albumin (HSA) were studied using fluorescence spectroscopy in combination with ultraviolet (UV) absorption spectroscopy, circular dichroism (CD) spectroscopy, Fourier transform infrared (FTIR) spectroscopy, and the molecular modeling method. UPT exhibited a strong ability to quench the intrinsic fluorescence of both BSA and HSA through a static quenching procedure. The binding constants of UPT and BSA or HSA were determined at different temperatures based on the relevant fluorescence data. The binding sites were obtained and the acting force was suggested to be mainly hydrophobic interaction, which was consistent with the result of the molecular modeling study, and there were also a number of hydrogen bonds between UPT and HSA. The results of determination of the proteins in bovine serum or human serum by this method were very close to those obtained by using Coomassie Brilliant Blue G-250 colorimetry. A practical method was proposed for the determination of UPT in bovine serum or human serum samples with satisfactory results. PMID- 16721792 TI - AFM force spectroscopy of the fibrinogen adsorption process onto dental implants. AB - Protein binding to implants is governed by the physicochemical properties of the biomaterial surface. The adhesion of a protein onto a solid surface is nonspecific. The aim of this study was to assess the adsorption process of fibrinogen at two different dental implants. The first biomaterial has a sand blasted titanium surface, whereas the second one is covered by a calcium phosphate coating. After scanning electron microscopy and atomic force microscopy characterization of the implant surfaces, force spectroscopy has been used to determine the unbinding force of fibrinogen adsorbed at the two different substrates. Force-measurement findings indicate that the detachment force of fibrinogen adsorbed onto both surfaces varies as a function of the interaction time. The mean strength of the unbinding forces increases with the interaction time (100 and 1,000 ms, respectively). However, experimental data suggest that fibrinogen fixes to the two studied biomaterials by different mechanisms. Moreover, it appears that, after an interaction time of 1,000 ms, the detachment force of the adsorbed protein is quite larger for the titanium surface than for the calcium phosphate coating. PMID- 16721793 TI - Prediction of tablet hardness based on near infrared spectra of raw mixed powders by chemometrics. AB - The purpose of this research is to elucidate the effect of lubricant mixing on tablet hardness by near-infrared (NIR) chemometrics as a basic study of process analytical technology. Formulation cellulose (F-C) consisted of sulpyrine (SP), microcrystalline cellulose (MC), and magnesium stearate (MgSt). Formulation lactose/starch (F-L) consisted of SP bulk drug powder, spray-dried lactose (SL), corn starch (CS), and MgSt. First, F-L and F-C without MgSt were mixed in a twin shell mixer for 60 min. MgSt was added to the mixed powder, and was mixed for various mixing times, after which the mixed powders were compressed by 8-mm diameter punch and die. NIR spectra of raw mixed powders of F-L and F-C were taken using a reflection type of Fourier transform NIR spectra spectrometer, and chemometric analysis was performed using principal component regression (PCR). The tablet hardnesses of F-L and F-C decreased with increasing mixing time. All NIR spectra of the mixed powders of F-L and F-C fluctuated depending on mixing time. In order to predict tablet hardness before tablet compression, NIR spectra of F-L and F-C mixed powders were analyzed and evaluated for hardness by PCR. The minimum standard error of cross-validation values could be realized by using five and six-principal component models, respectively. In the cases of F-L and F-C, the relationships between the actual and predicted tablet hardnesses showed straight lines, respectively. In the regression vectors of F-L and FC, the peaks related to hydrogen groups of SP, CS, and MC appeared as positive peaks. In contrast, the peaks related to hydrocarbon due to MgSt appeared as negative peaks in the regression vectors. The calibration models to evaluate the tablet hardness were obtained based on NIR spectra of raw mixed powders by PCR. This approach to predicting tablet hardness prior to compression could be used as a routine test to indicate the quality of the final product without spending time and energy to produce samples of questionable quality. PMID- 16721794 TI - Molecular vibrational analysis and MAS-NMR spectroscopy study of epilepsy drugs encapsulated in TiO2-sol-gel reservoirs. AB - A nanostructured matrix, consisting of titania, was designed in such a way that an antiepileptic drug could be encapsulated and released according to a well defined time release schedule. The titania was synthesized by a sol-gel method in which titanium n-butoxide was used as the precursor for the formation of the sol. The synthesis was optimized to yield a homogeneous particle size with a high porosity and an anatase crystal structure. The antiepilectic drugs, phenytoine or valproic acid, were added during the gelation stage in order to obtain a homogeneous gel phase. The resulting nanostructured matrix including the drug showed only weak attractive forces, such as London forces, dipole-dipole coupling, and in some cases hydrogen bonds. The resulting assembly, referred to as a reservoir, was characterized using conventional FTIR and NMR spectroscopic techniques. Theoretical simulation studies were performed so as to obtain an understanding of the equilibrium electrostatic potential distribution and the relative charges on the titania and the anticonvulsants. PMID- 16721795 TI - Impact of freeze-drying on ionization of sulfonephthalein probe molecules in trehalose-citrate systems. AB - "pH memory," i.e., correlation between pH of solution before freeze-drying and chemical reactivity in the freeze-dried state, has been reported in many systems. In this study, the "pH memory" is explored by comparing the extent of protonation of sulfonephthalein probe molecules, bromophenol blue, bromocresol green, and chlorophenol red, in aqueous solution in the pH range of 3.4-6.0 and in the resulting freeze-dried amorphous matrix (lyophile) containing trehalose and sodium citrate buffer. The protonation of the probe molecules was measured in the lyophiles by diffuse reflectance visible spectroscopy, and compared with that in the solution before drying. The protonation of the indicators in the amorphous matrix correlated with solution pH, that is, an increase in solution pH resulted in a progressive decrease in the indicator protonation in the corresponding lyophile. However, the protonation was consistently higher in the lyophile than in the corresponding solution. The Hammett acidity function of lyophiles was calculated based on the extent of protonation of the probe molecules. Protonation of the probe molecules and the Hammett acidity function depended not only on prelyophilization solution pH, but also on the residual water content and the presence of amorphous sugar in the lyophile. PMID- 16721796 TI - Stability profiles of drug products extended beyond labeled expiration dates. AB - The American Medical Association has questioned whether expiration dating markedly underestimates the actual shelf life of drug products. Results from the shelf life extension program (SLEP) have been evaluated to provide extensive data to address this issue. The SLEP has been administered by the Food and Drug Administration for the United States Department of Defense (DOD) for 20 years. This program probably contains the most extensive source of pharmaceutical stability data extant. This report summarizes extended stability profiles for 122 different drug products (3,005 different lots). The drug products were categorized into five groups based on incidence of initial extension failures and termination failures (extended lot eventually failed upon re-testing). Based on testing and stability assessment, 88% of the lots were extended at least 1 year beyond their original expiration date for an average extension of 66 months, but the additional stability period was highly variable. The SLEP data supports the assertion that many drug products, if properly stored, can be extended past the expiration date. Due to the lot-to-lot variability, the stability and quality of extended drug products can only be assured by periodic testing and systematic evaluation of each lot. PMID- 16721797 TI - Isolation and characterization of UHMWPE wear particles down to ten nanometers in size from in vitro hip and knee joint simulators. AB - There is currently considerable interest in the wear debris and osteolytic potential of different types of bearings used in total joint replacements. The biological activity of the wear debris is dependent on the size and volume of the particles produced. Wear volume also plays an important role in the functional biological activity of a joint replacement. In vitro studies have shown that crosslinking of ultra high molecular weight polyethylene (UHMWPE) acetabular cups and tibial trays produces a reduction in wear volume, and crosslinking has now been introduced clinically for both types of prostheses. Previous studies have identified both micron and submicron-sized polyethylene wear particles. The aim of this study was to characterize the wear and wear particles generated from moderately crosslinked GUR 1,020 GVF UHMWPE acetabular cups and tibial trays in hip and knee joint wear simulators down to 10 nanometers in size. The wear rates of the two prosthesis types were very similar at 25.6 +/- 5.3 mm(3) per million cycles for the hip prostheses and 22.75 +/- 5.95 mm(3) per million cycles for the knee prostheses. Nanometer-sized wear particles were isolated and characterized from both hip and knee simulator lubricants for the first time. Significantly higher numbers (p < 0.05) of particles in the nanometer (<0.1 microm) size range were produced by the hip prostheses compared to the knee prostheses. The knee prostheses produced larger particles, with the mode of particle size in the 0.1 1.0 microm size range, compared to <0.1 microm size range for the hip prostheses. In addition, the knee prostheses produced a greater volumetric concentration of wear particles in the 1.0-10 microm size range, and consequently lower specific biological activity and functional biological activity indices. These results indicated that the knee prostheses had a lower osteolytic potential compared to the hip prostheses. PMID- 16721798 TI - In vitro testing of calcium phosphate (HA, TCP, and biphasic HA-TCP) whiskers. AB - Calcium phosphate [single-phase hydroxyapatite (HA, Ca(10)(PO(4))(6)(OH)(2)), single-phase tricalcium phosphate (beta-TCP, Ca(3)(PO(4))(2)), and biphasic HA TCP] whiskers were formed by using a novel microwave-assisted molten salt mediated process. Aqueous solutions containing NaNO(3), HNO(3), Ca(NO(3))(2) x 4H(2)O, and KH(2)PO(4) (with or without urea) were used as starting reagents. These solutions were irradiated in a household microwave oven for 5 min. As recovered precursors were then simply stirred in water at room temperature for 1 h to obtain the whiskers of the desired calcium phosphate (CaP) bioceramics. These whiskers were evaluated, respectively, in vitro by (1) soaking those in synthetic body fluid (SBF) solutions at 37 degrees C for one week, and (2) performing cell attachment and total protein assay tests on the neat whiskers by using a mouse osteoblast cell line (7F2). beta-TCP, HA, and HA-TCP biphasic whiskers were all found to possess apatite-inducing ability when soaked in SBF. SBF-soaked whiskers were found to have BET surface areas ranging from 45 to 112 m(2)/g. Although the osteoblast viability and protein concentrations were found to be the highest on the neat HA whiskers, cells were attached and proliferated on all the whiskers. PMID- 16721799 TI - Biological performance of a new beta-TCP/PLLA composite material for applications in spine surgery: in vitro and in vivo studies. AB - The objective of this research was to carry out an in vitro and in vivo study of the biological performance of PLLA/beta-TCP composite materials, to estimate the scope of their potential applications in bone surgery. Samples with increasing beta-TCP (0-60% w/w) contents were processed by injection molding. The in vitro study consisted of an evaluation of inflammatory potential by assaying the IL 1alpha secreted by monocytes, and then cell proliferation (counting) and phenotype expression (PAL and I collagen) in human osteogenous cells. The in vivo study was carried out using cylindrical implants of composite materials composed of composite materials containing 0 or 60% beta-TCP and pure beta-TCP, respectively. The implants were inserted in femoral sites in rabbits, using the Kathagen protocol. Each animal received a 60% implant, with either a 0 or a 100% implant in the contralateral femur, so that the materials could be compared with one another. Five animals were examined for each material and implantation period, giving a total of 30 animals. This study showed that adding increasing percentages of beta-TCP to a lactic acid polymer matrix stimulated the proliferation of human osteogenous cells and synthesis of the extracellular bone matrix in a dose-dependent manner. In vivo results indicate that, in comparison with pure PLA, tricalcium phosphate-containing composite materials had faster degradation kinetics, caused less inflammatory reaction, and promoted contact osteogenesis. The composite material containing 60% beta-TCP demonstrated a similar performance to pure tricalcium phosphate bone grafts in terms of osteogenesis, and is apparently compatible with the production of intra-osseous implants for situations representing high levels of mechanical strain. PMID- 16721800 TI - The metabolic syndrome and risk of incident colorectal cancer. AB - BACKGROUND: The authors tested the hypothesis that the metabolic syndrome (> or =3 of the following components: high blood pressure, increased waist circumference, hypertriglyceridemia, low levels of high-density lipoprotein cholesterol, or diabetes/hyperglycemia) is a risk factor for colorectal cancer. METHODS: Data from the Atherosclerosis Risk in Communities (ARIC) multicenter prospective cohort study were used. Metabolic syndrome components and other risk factors were collected during 1987 to 1989 from the 14,109 men and women in these analyses. One hundred ninety-four incident colorectal cancers were identified through the Year 2000. Multivariate Cox proportional hazards regression analyses were used to examine associations. RESULTS: Baseline metabolic syndrome (> or =3 components vs. 0 components) had a positive association with age-adjusted and gender-adjusted colorectal cancer incidence (relative risk [RR], 1.49; 95% confidence interval [95%CI], 1.0-2.4); this association was attenuated after multivariate adjustment (RR, 1.39; 95%CI, 0.9-2.2). There was a dose-response association between colorectal cancer incidence and the number of metabolic syndrome components present at baseline (P for trend = .006) after multivariate adjustment. Analysis of gender revealed that the multivariate-adjusted association of metabolic syndrome with colorectal cancer was stronger in men (RR, 1.78; 95%CI, 1.0-3.6) and weaker in women (RR, 1.16; 95%CI, 0.6-2.2). CONCLUSIONS: In this population-based cohort, metabolic syndrome was a risk factor for incident colorectal cancer in men but not women. Evidence is growing that the metabolic syndrome may be a marker for a physiologic milieu of growth that encourages tumor initiation, promotion, and/or progression. PMID- 16721801 TI - Ewing sarcoma-family tumors that arise after treatment of primary childhood cancer. AB - BACKGROUND: Unlike osteosarcoma, the Ewing sarcoma family of tumors (ESFT) has rarely been reported as secondary malignant neoplasms after treatment of childhood cancer. ESFT arising as a second cancer was reviewed and characterized at our childhood cancer center. METHODS: A retrospective review was undertaken of 11,183 patients age <21 years who were treated for a primary cancer between March 1962 and December 2003 at St. Jude Children's Research Hospital. All cases of ESFT were confirmed to have a rearranged EWS gene. RESULTS: Six cases of ESFT (1.3% of 479 second cancers) were identified in patients previously treated for lymphoma (n = 3), leukemia (n = 1), retinoblastoma (n = 1), or Wilms tumor (n = 1). None of these patients had a family history suggestive of a familial cancer syndrome. The median time between diagnosis of primary cancer and diagnosis of ESFT was 5.9 years (range, 3.1-18.3 years). ESFT occurred in typical anatomic locations: rib (n = 2), chest wall soft tissues (n = 2), pelvis (n = 1), and extremity (n = 1). One tumor arose at the margin of a previous radiotherapy field and 1 arose distant from previous radiotherapy fields; all other patients had not received radiotherapy. Three patients are alive at the time of this report, including 2 whose ESFT was diagnosed more than 8 years ago. CONCLUSIONS: ESFT occurs rarely after treatment of a primary cancer during childhood, and most cases do not appear to be related to radiation therapy. Long-term survival can be achieved in some patients, and therefore secondary ESFT should be treated with curative intent. PMID- 16721802 TI - Transdisciplinary studies of surgical oncology trial accrual: A National Institutes of Health/National Cancer Institute Roadmap-affiliated project update. AB - BACKGROUND: A key objective of the National Institutes of Health Roadmap Initiative during the past 2 years has been to address scientific problems and issues through innovative research collaborations that cut across disciplines, traditions, and agendas. METHODS: Recognizing an opportunity to focus on the growing problem of low accrual to cancer surgical trials, the authors organized a multidisciplinary network of 21 clinical researchers and social/behavioral scientists to meet, review the issues, and organize working groups to study the problem. The authors, as representatives of the Karmanos Cancer Institute (Detroit, MI) and the American College of Surgeons Oncology Group, were awarded a Roadmap-affiliated grant from the National Cancer Institute under the "Meetings and Networks for Methodological Development in Interdisciplinary Research" funding mechanism. RESULTS: The overarching objective that guided the methodological developments of this network was to apply state-of-the-science methodologies and analytical strategies to examine the biobehavioral and interpersonal factors that facilitate or impede patients' treatment decision making. CONCLUSIONS: In this brief report, the authors describe the objectives of their multidisciplinary network and their progress to date. PMID- 16721803 TI - Low rates of colorectal, cervical, and breast cancer screening in Asian Americans compared with non-Hispanic whites: Cultural influences or access to care? AB - BACKGROUND: Asian Americans have lower cancer screening rates compared with non Hispanic whites (NHWs). Little is known about mechanisms that underlie disparities in cancer screening. The objectives of the current study were 1) to determine the relation between nativity, years in the United States, language, and cancer screening in NHWs and Asian Americans, independent of access to care and 2) to determine whether Asians reported different reasons than NHWs for not obtaining cancer screening. METHODS: This population-based study included 36,660 NHWs, 1298 Chinese, 944 Filipinos, 803 Koreans, 857 Vietnamese, and 1036 Other Asians from the 2001 California Health Interview Survey. The main study outcomes were 1) self-reported colorectal, cervical, and breast cancer screening and 2) reasons for not obtaining cancer screening. RESULTS: After adjusting for access to care, several Asian subgroups still had significantly lower rates of all types of cancer screening compared with NHWs. Adjusting for nativity, years in the United States, and English language attenuated the relation between Asian ethnicity and lower rates of colorectal and breast cancer screening. When they were asked what the most important reason was for not having each screening test, foreign-born Asians were significantly more likely than United States-born NHWs to report that they "didn't have problems/symptoms" (P < .01). CONCLUSIONS: Nativity, years in the United States, and English language may be markers of cultural differences that are mediating cancer screening disparities. Foreign born Asians may believe that cancer screening is in response to symptoms rather than tests that are used prior to the development of symptoms. Health education messages must consider how to communicate effectively that "cancer screening is valuable, because it finds cancer before it is advanced enough to cause symptoms." PMID- 16721804 TI - Histopathologic assessment of hot-spot microvessel density and vascular patterns in glioblastoma: Poor observer agreement limits clinical utility as prognostic factors: a translational research project of the European Organization for Research and Treatment of Cancer Brain Tumor Group. AB - BACKGROUND: Hot-spot microvessel density (MVD) and vascular patterns have been reported as histopathologic factors that influence prognosis in retrospective series of malignant gliomas. To investigate clinical utility, the authors systematically studied observer agreement on MVD and vascular patterns and the influence of repeatedly assessed data on patient outcomes in 2 independent glioblastoma series. METHODS: MVD and vascular patterns were assessed retrospectively by 5 observers in 1) a retrospectively compiled glioblastoma series that included 110 patients and 2) a glioblastoma series that included 233 patients who were treated within a randomized trial. MVD was determined in the field of greatest density ("hot-spot"). Predominantly classic or bizarre vascular patterns were determined by using a previously defined algorithm. RESULTS: Observer agreement on MVD was highly variable (range of kappa values, 0.464 0.901). The worst observer agreement was achieved when both the selection of hot spots and MVD counts were performed independently. Survival analysis did not show a consistent association between repeatedly assessed MVD and patient outcome. Observer agreement on vascular patterns was poor (kappa = 0.297). Survival analysis did not show a consistent association between repeatedly assessed vascular patterns and patient outcome. CONCLUSIONS: Observer agreement on hot spot MVD and vascular patterns in patients with glioblastoma was poor in independent assessments. MVD and vascular patterns were not associated consistently with patient outcome. Based on these findings, the authors concluded that poor observer agreement limits the clinical utility of histopathologically assessed hot-spot MVD and vascular patterns as prognostic factors in patients with glioblastoma. Improved methodologies for morphologic assessment of glioblastoma vascularization need to be identified. PMID- 16721805 TI - Cytomorphologic features of papillary lesions of the male breast: a study of 11 cases. AB - BACKGROUND: Breast masses occur in men far less commonly than women and are infrequently subjected to fine-needle aspiration (FNA) biopsy. Papillary lesions of the male breast are rare and are comprised of a spectrum of lesions ranging from papillary hyperplasia in gynecomastia to invasive papillary carcinoma. The following study describes the cytomorphology of papillary breast lesions in 11 men. The patients ranged in age from 23 to 78 years old and each presented with an unilateral subareolar or periareolar breast mass that varied in size from 0.5 to 3 cm. Two patients presented with bloody nipple discharge. METHODS: Archival material (8-year period) from FNA biopsies of papillary lesions of the male breast was reviewed. The reviewed cases were correlated with appropriate clinicopathologic follow-up. RESULTS: The smears had variable cellularity but all showed papillary clusters of mammary epithelial cells with and without fibrovascular cores. Single epithelial cells with a high nuclear-to-cytoplasmic ratio and eccentric nuclei were seen in all smears; however, these were more numerous in cases of adenocarcinoma. Hemosiderin-laden macrophages were present in all cases. Nipple discharge was seen only in the 2 benign lesions. All adenocarcinomas occurred in older men. CONCLUSIONS: The only cytologic criteria that differentiated benign from malignant papillary lesions were marked cellularity and the presence of abundant 3-dimensional clusters. To the best of the authors' knowledge, the current series is the largest in the English literature to date that examines the cytomorphologic features of papillary breast lesions in men. PMID- 16721806 TI - Occurrence of D-aspartate in the harderian gland of Podarcis s. sicula and its effect on gland secretion. AB - High concentrations of free D-aspartate (D-Asp), an amino acid well known for its neuroexcitatory activity, are endogeneously present in the Harderian gland (HG) of the lizard Podarcis s. sicula. This orbital gland consists of two different parts: the medial part, which is prevalently a mucous acinar gland, and the lateral part, which is a serous tubulo-acinar gland. To determine the physiological effect of D-Asp on exocrine secretion in HG, D-Asp (2.0 micromol/g b.w.) was injected intraperitoneally into lizards. We found that highest accumulations of exogenous D-Asp in HGs occurred 15 hr after the injection. Specifically, exogenous D-Asp prevalently stimulated serous secretion from the lateral portion of the gland, where immunohistochemical analysis revealed a major accumulation. Similarly, in the medial part of the gland, highly sulfated mucosubstances were observed after D-Asp injection. Further, in both parts of the HG, the electron microscope revealed euchromatic nuclei, a prominent rough endoplasmic reticulum, as well as numerous secretory granules within the acinar cells. Thus, following D-Asp injection, a 60% increase in HG total protein was detected. In addition, exogenous D-Asp induced changes in the electrophoretic pattern of HG. In conclusion, although further investigations are still needed to clarify the molecular pathway induced by D-Asp in exocrine secretion, this study does indicate that free D-Asp plays a significant role in the secretory activity of this gland. PMID- 16721807 TI - Differential expression of selected mitochondrial genes in hibernating little brown bats, Myotis lucifugus. AB - High rates of non-shivering thermogenesis by brown adipose tissue accompanied by additional shivering thermogenesis in skeletal muscle provide the powerful reheating of body organs that allows hibernating mammals to return from their state of cold torpor back to euthermic function. Previous studies have suggested that changes to brown adipose mitochondria occur during hibernation and are partially responsible for its capacity for non-shivering thermogenesis. The current study shows that selected mitochondrial enzyme activities are elevated and selected genes and proteins are induced during torpor in brown adipose tissue of the little brown bat, Myotis lucifugus. Cytochrome oxidase activity in brown adipose tissue was more than 3-fold higher during torpor than in euthermic animals. Transcript levels of mitochondria-encoded genes, coxII and nad4, were also 3-4-fold higher during torpor, as evidenced by northern blotting. By contrast, transcripts of these genes were unchanged in skeletal muscle during torpor. Protein levels of carnitine palmitoyl transferase-1beta, an enzyme embedded in the outer membrane of the mitochondria that is the rate-limiting step enzyme in beta-oxidation, were also elevated by 2-fold during torpor in brown adipose but were unchanged in skeletal muscle. Cloning and sequencing of a 624 bp segment of cpt-1beta revealed a number of amino acid substitutions in the bat protein as compared to CPT-1beta from other mammals; these may be beneficial for enzyme function at low body temperatures during torpor. This study provides further evidence for a key role of mitochondria in hibernation. PMID- 16721808 TI - High efficiency of meiotic gynogenesis in sea lamprey Petromyzon marinus. AB - Induction of androgenesis and gynogenesis by applying a pressure (PS) or heat shock (HS) to double the haploid chromosomal set results in progenies possessing only chromosomes from a single parent. This has never been accomplished in representatives of Agnatha. The objective of this study was to induce gynogenesis and androgenesis in sea lamprey Petromyzon marinus. For gynogenesis experiments, ultraviolet (UV)-irradiated sperm was used to activate sea lamprey eggs and HS or PS were applied to inhibit the second meiotic division and consequently induce diploidy in the embryos. The UV irradiation of immobilized sperm was performed for 1 min at 1,719 J m(-2). HS of 35+/-1 degrees C for 2 min and PS of 9,000 psi for 4 min were applied at different times after egg activation (8, 12, 20, and 24 min or 8, 16, and 24 min for HS or PS, respectively). Regardless of the induction time of the HS, survivals at pre-hatching stage were similar. In contrast, PS applied 8 min after activation appears to increase survival rate of pre-hatched embryos in comparison to 16 and 24 min after activation. In control groups, without shock treatment (no diploidization), there were no survivors. All deformed, gynogenetic embryos were confirmed to be haploids and died prior to burying themselves in the sand. We confirmed by flow cytometry that progenies produced using both shock methods surviving to the next stage, burying in the substrate, were diploid gynogenetic. For the androgenesis experiments, UV irradiated eggs (1,719 J m(-2) for 1 min) were fertilized with non-treated sperm and HS was applied to restore diploidy of the eggs. Several attempts have been made to optimize the parameters used. HS of 35+/-1 degrees C was applied 110, 140, 170, 200, and 230 min after activation for 2 min. Low yields of androgens were obtained and all animals died within a week after hatching. These techniques will allow to establish meiotic gynogenetic lines of sea lamprey for determining sex differentiation in this species and to analyze its hormonal and environmental regulation. PMID- 16721809 TI - Transcriptional census of 36 microdissected colorectal cancers yields a gene signature to distinguish UICC II and III. AB - UICC stage II and III colorectal cancers (CRC) differ fundamentally in prognosis and therapeutic concepts. To analyze differential gene expression between both stages and to establish a relationship between molecular background and clinical presentation, tumor material from 36 unselected consecutive patients presenting with sporadic CRC, 18 UICC stage II and 18 UICC stage III, were laser microdissected to separate epithelial tumor cells. Gene expression levels were measured using U133A Affymetrix gene arrays. Twelve CRC associated signal transduction pathways as well as all 22,000 probe sets were screened for differential gene expression. We identified a signature consisting of 45 probe sets that allowed discrimination between UICC stage II and stage III with a rate of correct classification of about 80%. The most distinctive elements in this signature were the gene GSTP-binding elongation factor (GSPT2) and the transcription factor HOXA9. Differential expression of these genes was confirmed by quantitative real-time polymerase chain reaction (p(HOXA9) = 0.04, p(GSTP2) = 0.02). Despite the reliability of the presented data, there was no substantial differential expression of genes in cancer-related pathways. However, the comparison with recently published data corroborates the 45 gene signature showing structural agreement in the direction of fold changes of gene expression levels for our set of genes chosen to discriminate between both stages. PMID- 16721810 TI - H-Ras selectively up-regulates MMP-9 and COX-2 through activation of ERK1/2 and NF-kappaB: an implication for invasive phenotype in rat liver epithelial cells. AB - One of the most frequent events in carcinogenesis is uncontrolled activation of Ras signaling pathway. A previous study demonstrated that the introduction of H Ras into the normal WB-F344 rat liver epithelial (WB) cell line and adult male F344 rats resulted in tumorigenicity. The present study investigated whether H Ras induced the invasive and migrative phenotypes in WB cells, and subsequently aimed at characterizing the underlying mechanisms. H-Ras induced the invasive and migrative phenotypes of WB cells with a selective up-regulation of matrix metalloproteinase (MMP)-9, but not MMP-2. Cyclooxygenase (COX)-2 and the subsequent production of prostaglandin E2 (PGE2) were also induced by H-Ras. Treatment of H-Ras WB cells with GM6001 and NS398, the inhibitors of MMPs and COX 2, respectively, significantly inhibited the H-Ras-induced invasive and migrative phenotypes. DNA binding activity of nuclear factor (NF)-kappaB, but not that of activator protein (AP)-1, was increased by H-Ras. Caffeic acid phenethyl ester and Bay 11-7082, specific inhibitors of NF-kappaB and IKK, respectively, significantly inhibited the expression of MMP-9 and COX-2, invasion and migration of H-Ras WB cells, revealing NF-kappaB as a transcriptional factor responsible for H-Ras-induced malignant phenotypic conversion of WB cells. Activation of ERKs pathway was critical for H-Ras-induced invasive and migrative phenotypes, up regulation of MMP-9 and COX-2 as well as enhanced DNA binding activity of NF kappaB in WB cells. Taken together, these results demonstrate that H-Ras up regulates MMP-9 and COX-2 through activation of ERKs and IKK-IkappaBalpha-NF kappaB signal pathway which may contribute to the malignant progression of WB rat liver epithelial cells. PMID- 16721811 TI - Expression of xenobiotic and steroid hormone metabolizing enzymes in human breast carcinomas. AB - The potential to metabolize endogenous and exogenous substances may influence breast cancer development and tumor growth. Therefore, the authors investigated the protein expression of Glutathione S-transferase (GST) isoforms and cytochrome P450 (CYP) known to be involved in the metabolism of steroid hormones and endogenous as well as exogenous carcinogens in breast cancer tissue to obtain new information on their possible role in tumor progression. Expression of GST pi, mu, alpha and CYP1A1/2, 1A2, 3A4/5, 1B1, 2E1 was assessed by immunohistochemistry for primary breast carcinomas of 393 patients from the German GENICA breast cancer collection. The percentages of positive tumors were 50.1 and 44.5% for GST mu and CYP2E1, and ranged from 13 to 24.7% for CYP1A2, GST pi, CYP1A1/2, CYP3A4/5, CYP1B1. GST alpha was expressed in 1.8% of tumors. The authors observed the following associations between strong protein expression and histopathological characteristics: GST expression was associated with a better tumor differentiation (GST mu, p = 0.018) and with reduced lymph node metastasis (GST pi, p = 0.02). In addition, GST mu expression was associated with a positive estrogen receptor and progesterone receptor status (p < 0.001). CYP3A4/5 expression was associated with a positive nodal status (p = 0.018). Expression of CYP1B1 was associated with poor tumor differentiation (p = 0.049). Our results demonstrate that the majority of breast carcinomas expressed xenobiotic and drug metabolizing enzymes. They particularly suggest that GST mu and pi expression may indicate a better prognosis and that strong CYP3A4/5 and CYP1B1 expression may be key features of nonfavourable prognosis. PMID- 16721812 TI - Prediction of recurrence in Ta urothelial cell carcinoma by real-time quantitative PCR analysis: a microarray validation study. AB - Accurate prediction of tumor recurrence in patients with superficial urothelial cell carcinoma (UCC) might result in a significant reduction of invasive follow up cystoscopies. A recent study identified a panel of 26 genes from a large cDNA microarray analysis of bladder tumors that discriminated between early- and late recurring patients with superficial Ta tumors (Dyrskjot et al., Nat Genet 2003;33:90-6). We aimed to validate this panel of genes in 44 primary Ta UCCs (23 and 21 tumors from patients with short or prolonged recurrence-free periods, respectively), by real-time quantitative PCR. Statistical analysis showed marginal significant different mRNA expression levels between the 2 patient groups. To evaluate a supplementary effect of genes for the identification of patients with short or prolonged recurrence-free intervals, forward logistic regression analysis was applied. This revealed that a combination of the expression profiles of the genes HNRPK, LTB4DH and ANP32B resulted in the best performance, although the combination only marginally increased the predictive value of HNRPK alone. Comparing the receiver-operating-characteristic curves for HNRPK expression among patients with short or prolonged recurrence-free periods, revealed an area under the curve of 0.696 (95% CI, 0.537-0.855). Using the median HNRPK expression level as cut-off, a sensitivity of 69.6% and a specificity of 71.4% were obtained for the identification of patients with short or prolonged recurrence-free periods, respectively. In conclusion, we were not able to confirm the microarray gene expression pattern of the 26 genes shown by Dyrskjot et al. The discovery of accurate recurrence predictive markers, therefore, remains a challenge. PMID- 16721813 TI - DNA mismatch repair as an effector for promoting phorbol ester-induced apoptotic DNA damage and cell killing: implications in tumor promotion. AB - Phorbol ester was known to activate protein kinase C (PKC) and exert numerous cellular effects, including proliferation, apoptosis, and oncogenic transformation. How phorbol ester stimulates both apoptosis and tumor promotion is not clear. Here DNA mismatch repair (MMR)-proficient human colon cancer cells (DLD-1+Ch2; hMSH6+) treated with 12-O-tetradecanoylphorbol-13-acetate (TPA) undergo rapid cell death, which is significantly abolished by staurosporine (PKC inhibitor) or antioxidant, compared with the paired MMR-deficient (DLD-1; hMSH6-) cells. Induction of reactive oxygen species (ROS) by TPA is shown to be one of downstream effectors required, but not sufficient, for cell killing as it is also observed in DLD-1 cells. Strikingly, DLD-1+Ch2 cells selected for resistance to TPA are found to lose the expression of hMSH6. Treatment of TPA-resistant DLD 1+Ch2 cells with 5-aza-2'-deoxycytidine, not only restores hMSH6 expression but also resensitizes TPA-resistant cells to TPA, suggesting that expression of hMSH6 is transcriptionally silenced by cytosine methylation confirmed directly by bisulfite sequencing. Knockdown hMSH6 or hPMS2 with siRNA in DLD-1+Ch2 cells resulted in more resistant to TPA-induced cell killing, further suggesting that MMR proteins involve in TPA or ROS-induced cell killing. Results suggest that deficiency in MMR could promote tumorigenesis by inhibiting apoptotic responses to ROS-mediated DNA damages as ROS are continuously produced as a byproduct of normal metabolism. PMID- 16721814 TI - Reduced incidence of cervical cancer in mothers of sons with allergic rhinoconjunctivitis, asthma or eczema. AB - Because infection with human papillomavirus is a necessary cause of cervical cancer, it is likely that host immunological factors involved in defense against such infections are important for susceptibility to this cancer. By examining associations between allergy in sons and cervical cancer in their mothers, we aimed to test for genetic components involved in both allergy and cervical cancer development. Women born in Sweden between 1932 and 1960 with at least 1 son with medical records from military conscription examination were included in the study (N = 717,963). Among these women there were 41,910 in situ and 3,618 invasive cases of cervical cancer. Hazard ratios of in situ and invasive cervical cancer were estimated as functions of allergic rhinoconjunctivitis, asthma and eczema diagnoses in sons. Adjustment was made for the possible confounders: age, year of birth, education, socio-economic index, geography, number of conscripted sons and total number of offspring. The risk of in situ and invasive cervical cancer was lower for women having sons diagnosed with hypersensitivity (allergic rhinoconjunctivitis, asthma or eczema). Fully adjusted hazard ratios for women with 1 hypersensitive son were for in situ 0.86 (95% CI 0.84-0.88) and for invasive cervical cancer 0.82 (95% CI 0.74-0.91). The protective effects were similar between the 3 allergic diagnoses and increased with number of sons with a diagnosis. There was no significant association between non-cervical cancer in mothers and allergy in sons. These results strengthen the hypothesis that inherited immunological factors are important in determining risk of cervical cancer, probably by affecting mechanisms for viral persistence. PMID- 16721815 TI - The assessment of levator trauma: a comparison between palpation and 4D pelvic floor ultrasound. AB - AIMS: Major morphological abnormalities of the pubovisceral muscle are observed in 10-20% of women symptomatic of pelvic floor disorders. Such defects arise in childbirth and are associated with prolapse. While they are clearly evident on 3D ultrasound and MR imaging, such defects can be difficult to detect clinically. We intended to compare findings on palpation with the results of ultrasound imaging. MATERIAL AND METHODS: Fifty-five women were recruited in a prospective observational study and assessed by interview, vaginal examination by a trained pelvic floor physiotherapist, and 3D/4D pelvic floor ultrasound. The vaginal examination involved digital assessment of muscle strength (modified Oxford grading) and morphological abnormalities. The ultrasound examination involved acquisition of volume datasets taken at rest, on Valsalva and pelvic floor muscle contraction. Assessments were undertaken supine and after voiding. Ultrasound operator and physiotherapist were blind to each other's findings. RESULTS: A comparison of 3D ultrasound and palpation was possible in 54 cases. Average Oxford grading was weakly associated with reduction in hiatal dimensions on contraction (r = -0.32, P = 0.024). A marked increase in hiatal dimensions detected on palpation was associated with increased hiatal area on Valsalva (P = 0.027). Defects were observed in 7/54 (13%) on ultrasound and in 10/55 (18%) on palpation. There was poor agreement between the methods, with only two defects picked up equally by both methods. CONCLUSIONS: Palpation of the pubovisceral muscle correlates poorly with 3D/4D pelvic floor ultrasound imaging for the assessment of muscular defects. PMID- 16721816 TI - Predictive value of initial urodynamic pattern on urinary continence in patients with myelomeningocele. AB - AIMS: To investigate whether the initial urodynamic pattern may predict urinary continence and the need for adjunctive incontinence surgery in patients with myelomeningocele. PATIENTS AND METHODS: One hundred and twenty-three patients with myelomeningocele were included in our retrospective study. The patients were divided based on the urodynamic pattern at initial evaluation into four categories: overactive detrusor with overactive (spastic) sphincter (group 1, n = 43), overactive detrusor with underactive/acontractile sphincter (group 2, n = 37), underactive/acontractile detrusor with overactive (spastic) sphincter (group 3, n = 8), and underactive/acontractile detrusor with underactive/acontractile sphincter (group 4, n = 35). Urinary continence status at the last follow-up and the need for adjunctive incontinence surgery were compared between the four groups. RESULTS: Mean follow-up was 10 years and mean age at the last follow-up 17 years. A socially acceptable continence status (continent or socially dry) was achieved in 74% and was significantly different among the four groups (P = 0.023): 86% and 87% of the patients in group 1 and 3 became continent or socially dry, but only 57% and 74% of those in group 2 and 4, respectively. Although none of the patients in group 3 underwent adjunctive incontinence surgery compared to about 25% in the other groups, statistically the difference was not significant (P = 0.48). CONCLUSIONS: The initial urodynamic pattern is very useful in counseling families by predicting urinary continence in patients with myelomeningocele. The chances of becoming continent or at least socially dry are best for patients with overactive (spastic) sphincter. PMID- 16721817 TI - Role of fluorine-18 fluoro-deoxyglucose positron emission tomography scan in the evaluation and follow-up of patients with low-grade lymphomas. AB - BACKGROUND: Fluorine-18 fluoro-deoxyglucose positron emission tomography (FDG PET) scanning has excellent sensitivity and specificity for staging non-Hodgkin lymphomas, but to the authors' knowledge few studies to date have evaluated FDG PET in low-grade lymphomas only. METHODS: A retrospective study was performed on patients with biopsy-proven nontransformed and transformed follicular lymphoma (FL), B-cell small-cell lymphocytic lymphoma (SLL/CLL), or marginal zone lymphoma (MZL) who underwent PET and computed tomography (CT) scans within 3 weeks. Standard uptake values (SUV) of all abnormal foci were measured. RESULTS: In FL, PET demonstrated 94% sensitivity and 100% specificity for staging. PET was more specific than CT for detecting recurrence or assessing therapeutic responses (91% vs. 50%). FDG avidity among patients with WHO Grades 1, 2, and 3 disease was not significantly different (analysis of variance [ANOVA]). For MZL staging, PET had moderate sensitivity (71%) and outperformed CT alone in the depiction of extranodal sites (85% vs. 57% sensitivity). In SLL/CLL, PET sensitivity was 53% and underestimated disease extent in 5 of 19 patients (26%) compared with CT. PET did not affect initial management but confirmed suspected recurrences in 75% of patients. Nontransformed FL had a higher SUV (ANOVA, P < .05) compared with MZL and SLL/CLL. SUV was higher in transformed than in nontransformed tumors (P < .001, Student t test). CONCLUSIONS: PET usefulness in staging low-grade lymphomas varies depending on histology. PET sensitivity is excellent in FL and moderate in MZL. PET is more specific than CT for follow-up in all types. PET has limited usefulness for SLL/CLL staging. However, a suggestive pattern of hazy and mild uptake was often noted in positive scans. In all low-grade lymphomas, the emergence of foci of intense uptake should raise suspicion of conversion to high grade disease. PMID- 16721818 TI - Overexpression of Jun activation domain-binding protein 1 in nonsmall cell lung cancer and its significance in p27 expression and clinical features. AB - BACKGROUND: Decreased expression of p27, which is an inhibitor of cyclin dependent kinase, is associated with cancer aggressiveness. It is believed that Jun activation domain-binding protein 1 (Jab1) plays a role in p27 degradation in a manner that is independent from the role played by S-phase kinase-associated protein 2 (Skp2). To examine the clinical significance of Jab1 in nonsmall cell lung cancer (NSCLC), the protein expression of Jab1 in tumor tissues was investigated with regard to the expression of p27 and Skp2. METHODS: The clinicopathologic features and immunohistochemical expression levels of Jab1, p27, and Skp2 proteins were studied in 138 specimens from patients who underwent surgical resection for NSCLC. Survival analyses were performed by using the Kaplan-Meier method and Cox regression models. RESULTS: High Jab1 expression (>50% of cancer cell nuclei stained) was observed in 81 specimens (59%). In Skp2 negative specimens (n = 81), an inverse correlation between the protein expression of p27 and Jab1 was observed both with the chi-square test (P = .02) and with the Mann-Whitney U test (P = .02). High Jab1 expression levels were related to poor outcomes in 118 patients who underwent complete resection, with a 5-year overall survival rate of 43.9% for patients who had high Jab1 expression and 63.1% for patients who had low Jab1 expression (P = .01). This difference was greater in patients who had Skp2-negative specimens (51.9% vs. 79.3% (P = .02), but it was not significant in patients who had Skp2-positive specimens. The multivariate analysis revealed that Jab1 expression was an independent prognostic factor for survival in patients with NSCLC (relative risk, 2.247; P = .01). CONCLUSIONS: The current results showed for the first time that high Jab1 protein expression is related to poor outcome in patients with NSCLC and that this protein may be a target of therapy in NSCLC. PMID- 16721819 TI - High-dose cytosine arabinoside in the treatment of acute myeloid leukemia: Review of three randomized trials. AB - BACKGROUND: The use of high-dose cytosine arabinoside (HDAraC) during induction may improve outcomes in patients with acute myeloid leukemia (AML) compared with standard-dose AraC (SDAraC). The objective of this review was to assess the impact of HDAraC during induction therapy for patients with AML based on results from randomized trials. METHODS: All randomized trials in the field were identified by using a predefined search strategy. Trials that assessed the impact of HDAraC compared with SDAraC as induction therapy for adult patients with AML in a randomized fashion and that reported the relevant endpoints were included. Data were extracted from each trial by both reviewers according to prespecified criteria. RESULTS: No differences between HDAraC and SDAraC were found with regard to complete remission rates (relative risk, 1.00; 95% confidence interval [95% CI], 0.92-1.10). The weighted mean difference (WMD) for median recurrence free survival (RFS) was 4.19 in favor of HDAraC (95% CI, 0.59-7.78; P = .02). The WMD for 4-year RFS was 10.98 in favor of HDAraC (95% CI, 1.02-20.94; P = .03). The WMD for median overall survival (OS) was - 0.22 for HDAraC compared with SDAraC (95% CI, - 2.76-2.32; P = .9). Data regarding the median OS was heterogeneous between studies (chi-square P = .00), with 2 studies in favor of HDAraC and 2 studies in favor of SDAraC. The WMD for 4-year OS was 6.21 in favor of HDAraC (95% CI, 2.70-9.72; P = .0005). CONCLUSIONS: Induction therapy with HDAraC improved long-term disease control and overall survival in adults age < 60 years with de novo AML. It remains unknown whether patients should receive HDAraC during induction or if it is to be given during postremission therapy. Further analyses should focus on this issue and on the effects of HDAraC in prognostically different subgroups of patients with AML. PMID- 16721820 TI - Biological activity of the functional epitope of ciguatoxin fragment AB on the neuroblastoma sodium channel in tissue culture. AB - It is well established that the targeted receptor for ciguatoxin (CTX) in mammalian tissues is the sodium channel, affecting the influx of sodium into cells and altering the action potential and function of the cell. Since the syntheses of fragments of CTX has become available, our focus has been on the receptor functions of the west sphere AB and east sphere JKLM fragments using the neuroblastoma cell assay, guinea pig atrium assay, and the membrane immunobead assay (MIA). The data presented here suggest that the west sphere AB of the ciguatoxin molecule is the active portion and is responsible for the activation of the sodium channels. PMID- 16721821 TI - Impact of opportunistic Mycobacterium tuberculosis infection on the phenotype of peripheral blood T cells of AIDS patients. AB - While the detrimental consequences of opportunistic tuberculosis (TB) in the course and outcome of HIV-1 infection are well studied, little information about the impact of the mycobacterial infection on the phenotype of T lymphocytes is available. In this study we analyzed by cytofluorimetry the peripheral blood T cell phenotype of 13 patients with AIDS, 23 HIV-1 negative patients with active pulmonary TB, nine HIV-1/Mycobacterium tuberculosis coinfected individuals, and 21 age- and sex-matched healthy controls. CD4+ T cells were equally depleted in AIDS and coinfection (P<0.001). The findings suggest a rescuing effect of the added mycobacterial infection. CD3 T cell loss was not observed in coinfection, whereas it was severe in AIDS (P<0.001). Similar (albeit less striking) effects were observed with other markers (CD45RA, CD45RO, and CD27) that were diminished in CD4+ T cells of AIDS patients. Apparent detrimental effects of the added mycobacterial infection were the increased expression of the proapoptotic molecule CD95 on CD4+ T cells, and decreased expression of the major costimulatory molecule CD28 on CD8+ T cells. In this work we show that M. tuberculosis infection modifies the T cell phenotype of the HIV-1 infected individual. PMID- 16721822 TI - Vitamin D receptor gene polymorphisms are associated with risk of Hashimoto's thyroiditis in Chinese patients in Taiwan. AB - The etiology of the autoimmune thyroid, Hashimoto's thyroiditis (HT) is not very clear. However, genetic susceptibility is thought to play a critical role. The vitamin D receptor (VDR)-related endocrine system has been demonstrated to be able to carry out modulation of the immune response. Here, we investigated whether single nucleotide polymorphisms (SNPs) of VDR are associated with HT patients. VDR SNP was detected by polymerase chain reaction (PCR)-based restriction analysis in 109 patients with HT and 90 normal controls. Significant differences were found in the genotype distribution of VDR SNP between Hashimoto's thyroiditis patients and controls (P=0.0458). Allelic frequency of the VDR gene distinguished HT patients from controls (P=0.0089). The results revealed a significant difference between HT patients and normal controls in VDR SNP and a statistic correlation between VDR-FokI polymorphisms and HT formation. It could be concluded that patients who carry the C/C homozygote of the VDR-FokI gene polymorphism in exon 2 may have a higher risk of developing HT in Chinese patients in Taiwan. PMID- 16721823 TI - Functional overexpression and characterization of human bradykinin subtype 2 receptor in insect cells using the baculovirus system. AB - Bradykinin exerts its actions via binding to B1 and B2 receptors (B1R and B2R), which are members of G protein-coupled receptor superfamily. B2R is constitutively expressed in a variety of cells such as endothelial cells, vascular smooth muscle cells, and cardiomyocytes and it is an important drug target for the treatment of cardiovascular disorders. During this study, the human B2R was functionally overexpressed in insect cells using the baculovirus expression system. The maximum expression level in Sf9 cells under optimized condition was 10 pmol/mg. This corresponds to approximately 0.25 mg active receptor per liter culture. The recombinant receptor showed high affinity for its endogenous ligand bradykinin, similar to the B2R expressed in native tissues. Functional coupling of the recombinant receptor to the endogenous G alpha(s) protein was demonstrated via cAMP release assay upon agonist stimulation. Confocal laser scanning microscopy and immunogold-labeling experiment revealed that the recombinant B2R was mainly localized intracellularly and only a minor fraction of the recombinant receptor reached the plasma membrane. To our knowledge, this is the first report of high level expression of recombinant B2R in insect cells and provides a way for large scale production and structural characterization of this receptor. PMID- 16721824 TI - Amyloid beta oligomerization is induced by brain lipid rafts. AB - Detergent-resistant lipid rafts are required for the generation of Abeta as they concentrate not only amyloid precursor protein (APP), but also the beta- and gamma-secretase that convert APP to Abeta. Recently, Abeta has been shown to be oligomerized, which results in neuronal cytotoxicity and synaptic failure. In this study, we have demonstrated that Abeta oligomers appeared immediately after the incubation of Abeta with lipid rafts isolated from the brain tissues of rats, and were converted into few Abeta fibrils, even after longer periods of incubation. The oligomerization of Abeta was not abolished after the brain lipid rafts were treated with heat, or with protease K, implying that the lipid raft proteins were determined not to be prerequisites for Abeta oligomerization. The cholesterol present in the lipid rafts might not be essential to Abeta oligomerization because Abeta oligomerization was not prevented after the cholesterol was removed from the lipid rafts with methyl-beta-cyclodextrin (MbetaCD). The Abeta oligomerization was accelerated by the application of lipid rafts isolated from ganglioside-rich cells, C2C12 cells, whereas this was not observed with the lipid rafts isolated from ganglioside-poor cells SK-N-MC and HeLa cells. In addition, lipid raft-induced Abeta oligomerization was shown to be inhibited in CHO-K1 cells which were defective with regard to ganglioside biosynthesis. This indicates that Abeta oligomerization requires gangliosides that are enriched in the lipid rafts. PMID- 16721825 TI - Expression and regulation of resistin in osteoblasts and osteoclasts indicate a role in bone metabolism. AB - The adipose tissue is the site of expression and secretion of a range of biologically active proteins, called adipokines, for example, leptin, adiponectin, and resistin. Leptin has previously been shown to be expressed in osteoblasts and to promote bone mineralization, whereas adiponectin expression is enhanced during osteoblast differentiation. In the present study we explored the possible role of resistin in bone metabolism. We found that resistin is expressed in murine preosteoclasts and preosteoblasts (RAW 264.7, MC3T3-E1), in primary human bone marrow stem cells and in mature human osteoblasts. The expression of resistin mRNA in RAW 264.7 was increased during differentiation and seemed to be regulated through PKC- and PKA-dependent mechanisms. Recombinant resistin increased the number of differentiated osteoclasts and stimulated NFkappaB promoter activity, indicating a role in osteoclastogenesis. Resistin also enhanced the proliferation of MC3T3-E1 cells in a PKA and PKC-dependent manner, but only weakly interfered with genes known to be upregulated during differentiation of MC3T3-E1 into osteoblasts. All together, our results indicate that resistin may play a role in bone remodeling. PMID- 16721826 TI - Different signaling responses to anti-proliferative agents in human aortic and venous smooth muscle cells. AB - Proliferation of smooth muscle cells (SMCs) contributes to the stenosis of coronary arteries and vascular grafts. Local delivery of anti-proliferative drugs can prevent vascular stenosis. To understand the cellular responses to anti proliferative agents, we investigated the signaling events in cultured human aortic SMCs (ASMCs), saphenous venous SMCs (VSMCs), and dermal fibroblasts (DFs) in response to paclitaxel or etoposide. Cellular mitochondrial and proliferative activities were examined with the methylthiazoletetrazolium (MTT) dye reduction and the bromodeoxyuridine (BrdU) incorporation assay, respectively. Cell proliferation was almost completely suppressed by paclitaxel or etoposide, but apoptosis was achieved in only about 50% of cells at the highest drug concentrations, suggesting the presence of compensatory mechanisms to prevent apoptosis. Examination of three important signaling pathways revealed significant differences between ASMCs, VSMCs, and DFs. Treatment with either paclitaxel or etoposide caused a transient phosphorylation/activation of p42 MAPK in ASMCs and DFs, but had no effect on phospho-p42/44 MAPK in VSMCs. High-dose etoposide enhanced p38 MAPK activation in ASMCs, but not in VSMCs. The p38 inhibitor, PD169316, partially inhibited etoposide-induced ASMC apoptosis, but induced apoptosis in VSMCs. The effects of etoposide and paclitaxel on Akt also differed between ASMCs and VSMCs. These observations indicate that ASMCs and VSMCs differ in the response of signaling pathways to anti-proliferative agents. In ASMCs, p42/44 MAPK appears to serve a pro-survival role, whereas p38 MAPK is a pro apoptotic regulator. In contrast, p38 MAPK is an important pro-survival regulator in VSMCs and p42/44 MAPK appears to play a minor role in responding to anti proliferative drugs. PMID- 16721827 TI - CDC2L5, a Cdk-like kinase with RS domain, interacts with the ASF/SF2-associated protein p32 and affects splicing in vivo. AB - The human CDC2L5 gene encodes a protein of unknown physiological function. This protein is closely related to the cyclin-dependent kinase (Cdks) family and contains an arginine/serine-rich (RS) domain. The Cdks were first identified as crucial regulators of cell-cycle progression, more recently they were found to be involved in transcription and mRNA processing. RS domains are mainly present in proteins regulating pre-mRNA splicing, suggesting CDC2L5 having a possible role in this process. In this study, we demonstrate that CDC2L5 is located in the nucleoplasm, at a higher concentration in speckles, the storage sites for splicing factors. Furthermore, this localization is dependent on the presence of the N-terminal sequence including the RS domain. Then, we report that CDC2L5 directly interacts with the ASF/SF2-associated protein p32, a protein involved in splicing regulation. Overexpression of CDC2L5 constructs disturbs constitutive splicing and switches alternative splice site selection in vivo. These results argue in favor of a functional role of the CDC2L5 kinase in splicing regulation. PMID- 16721828 TI - Plasma membrane destination of the classical Xenopus laevis progesterone receptor accelerates progesterone-induced oocyte maturation. AB - Xenopus laevis oocyte maturation is induced by the steroid hormone progesterone through a non-genomic mechanism initiated at the cell membrane. Recently, two Xenopus oocyte progesterone receptors have been cloned; one is the classical progesterone receptor (xPR-1) involved in genomic actions and the other a putative seven-transmembrane-G-protein-couple receptor. Both receptors are postulated to be mediating the steroid-induced maturation process in the frog oocyte. In this study, we tested the hypothesis that the classical progesterone receptor, associated to the oocyte plasma membrane, is participating in the reinitiation of the cell cycle. Addition of a myristoilation and palmytoilation signal at the amino terminus of xPR-1 (mp xPR-1), increased the amount of receptor associated to the oocyte plasma membrane and most importantly, significantly potentiated progesterone-induced oocyte maturation sensitivity. These findings suggest that the classical xPR-1, located at the plasma membrane, is mediating through a non-genomic mechanism, the reinitiation of the meiotic cell cycle in the X. laevis oocyte. PMID- 16721829 TI - Expression of genes regulating malonyl-CoA in human skeletal muscle. AB - In humans and animal models, increased intramuscular lipid (IML) stores have been implicated in insulin resistance. Malonyl-CoA plays a critical role in cellular lipid metabolism both by serving as a precursor in the synthesis of lipids and by inhibiting lipid oxidation. In muscle, Malonyl-CoA acts primarily as a negative allosteric regulator of carnitine palmitoyl transferase-1 (CPT1) activity, thereby blocking the transport of long chain fatty acyl CoAs into the mitochondria for oxidation. In muscle, increased malonyl-CoA, decreased muscle CPT1 activity, and increased IML have all been reported in obesity. In order to determine whether malonyl-CoA synthesis might be under transcriptional as well as biochemical regulation, we measured mRNA content of several key genes that contribute to the cellular metabolism of malonyl-CoA in muscle biopsies from lean to morbidly obese subjects. Employing quantitative real-time PCR, we determined that expression of mitochondrial acetyl-CoA carboxylase 2 (ACC2) was increased by 50% with obesity (P < 0.05). In both lean and obese subjects, expression of mitochondrial ACC2 was 20-fold greater than that of cytoplasmic ACC1, consistent with their hypothesized roles in synthesizing malonyl-CoA from acetyl-CoA for CPT1 regulation and lipogenesis, respectively. In addition, in both lean and obese subjects, expression of malonyl-CoA decarboxylase was approximately 40-fold greater than fatty acid synthase, consistent with degradation, rather than lipogenesis, being the primary fate of malonyl-CoA in human muscle. No other genes showed signs of increased mRNA content with obesity, suggesting that there may be selective transcriptional regulation of malonyl-CoA metabolism in human obesity. PMID- 16721830 TI - Unloading-induced bone loss was suppressed in gold-thioglucose treated mice. AB - Loss of mechanical stress causes bone loss. However, the mechanisms underlying the unloading-induced bone loss are largely unknown. Here, we examined the effects of gold-thioglucose (GTG) treatment, which destroys ventromedial hypothalamus (VMH), on unloading-induced bone loss. Unloading reduced bone volume in control (saline-treated) mice. Treatment with GTG-reduced bone mass and in these GTG-treated mice, unloading-induced reduction in bone mass levels was not observed. Unloading reduced the levels of bone formation rate (BFR) and mineral apposition rate (MAR). GTG treatment also reduced these parameters and under this condition, unloading did not further reduce the levels of BFR and MAR. Unloading increased the levels of osteoclast number (Oc.N/BS) and osteoclast surface (Oc.S/BS). GTG treatment did not alter the basal levels of these bone resorption parameters. In contrast to control, GTG treatment suppressed unloading-induced increase in the levels of Oc.N/BS and Oc.S/BS. Unloading reduced the levels of mRNA expression of the genes encoding osteocalcin, type I collagen and Cbfa1 in bone. In contrast, GTG treatment suppressed such unloading-induced reduction of mRNA expression. Unloading also enhanced the levels of fat mass in bone marrow and mRNA expression of the genes encoding PPARgamma2, C/EBPalpha, and C/EBPbeta in bone. In GTG-treated mice, unloading did not increase fat mass and the levels of fat-related mRNA expression. These results indicated that GTG treatment suppressed unloading-induced alteration in bone loss. PMID- 16721831 TI - High levels of neopterin and interleukin-3 in sickle cell disease patients. AB - Sickle cell disease (SCD) is recognized as a chronic inflammatory condition. Cytokines are released in response to stress or pathological situations, and influence hematopoiesis. The aim of this study was to evaluate interleukin-3 (IL 3), interferon-gamma (IFN-gamma), and neopterin (NP) levels in steady-state patients with sickle cell anemia (SS) (n = 35) and SC hemoglobinopathy (n = 15) in order to verify the possible action of those cytokines and NP on iron metabolism and hematopoiesis. Serum IL-3 concentration was higher in SS and SC groups than in controls, whereas IFN-gamma levels did not differ among groups. SS patients presenting hemoglobin fetal (HbF) >or=8.5% had significantly higher IL-3 levels than those with HbF <8.5% (P = 0.0338). No correlation was observed among inflammatory and iron metabolism parameters. Significant correlations were observed between IL-3 and Hb levels (r = 0.4633, P = 0.0457), and IL-3 and HbF levels (r = 0.6011, P = 0.0065). A negative correlation was observed between IL-3 and reticulocyte counting (r = -0.4632, P = 0.0457) only in the SS group. NP levels were significantly higher in the SS and SC groups than in controls, but did not differ between SS and SC. No correlation was observed between NP and iron metabolism parameters. These data suggest that IL-3 stimulates hematopoiesis, and that SS patients, even in steady state, have macrophage/monocyte activation (represented by high levels of NP) that probably contributes to their chronic inflammatory condition. PMID- 16721832 TI - Study of V1a vasopressin receptor gene single nucleotide polymorphisms in platelet vasopressin responsiveness. AB - There is a significant heterogeneity among individuals in terms of platelet aggregation response to arginine vasopressin (AVP). The aim of this study was to evaluate whether four single nucleotide polymorphisms (SNPs) in the promoter region of vasopressin V1a receptor gene (V1aR) could be used as genetic markers for divergent platelet aggregation response to AVP. Seventeen of 33 subjects showed more than 60% of maximum platelet aggregation and were classified as responders. Sixteen were classified as nonresponders because they had less than 30% aggregation. In a preliminary study, V1aR gene sequences were determined in two responders and two nonresponders. We found four SNPs in the promoter region of the V1aR gene: -6951G/A, -4112A/T, -3860T/C, and -242C/T. In all 33 subjects the genotypes of four SNPs were determined using either polymerase chain reaction (PCR) with allele-specific primers or PCR followed by restriction-fragment length polymorphism (RFLP). There were no differences in the AVP-induced aggregation between the subjects with and without variant alleles of each four SNPs. The genotype frequencies of four SNPs of V1aR were almost identical between AVP responders and nonresponders. These results suggest that the four SNPs in the promoter region of the V1aR gene may not be useful as genetic markers for platelet aggregation heterogeneity. PMID- 16721833 TI - Natriuretic peptide Val7Met substitution and risk of coronary artery disease in Greek patients with familial hypercholesterolemia. AB - Atrial natriuretic peptide (ANP or NPPA) is the precursor protein of the form of amyloidosis called isolated atrial amyloid (IAA), which is related to the increased incidence of cardiac pathological conditions with age. Familial hypercholesterolemia (FH) patients are characterized by high concentrations of low-density lipoprotein cholesterol (LDL-C), which frequently gives rise to premature coronary artery disease (CAD). However, not all FH patients have the same clinical phenotype. The aim of the present study was to assess the relationship between ANP polymorphisms and apolipoprotein (Apo) A1 levels and CAD risk in FH patients. Transition T2238C, which leads to ANP with two additional arginines, and G664A (Val7Met) were investigated with lipid values and clinical phenotype in 83 FH patients. ApoA1 and HDL cholesterol levels were lower in GA patients compared to GG homozygotes for the G664A polymorphism. No association was found between the G664A polymorphism and CAD in our population. Moreover, ApoA1 and high-density lipoprotein cholesterol (HDL-C) levels did not differ among the different genotypes of the T2238C polymorphism, even after adjusting for age and sex. The 664A allele of the ANP polymorphism is associated with lower levels of ApoA1 and HDL-C in FH patients, but not with CAD risk. Concerning the T2238C polymorphism, no effect was found on lipid parameters or CAD incidence. PMID- 16721834 TI - High-density lipoprotein subfractions in normolipidemic individuals without clinical atherosclerosis lipoprotein subfractions in an adult population. AB - This study evaluated the serum concentrations of lipids, lipoproteins, apolipoproteins, and high-density lipoprotein (HDL) subfractions in Brazilian adults. We analyzed the distribution of lipids in HDL2 and HDL3 in a normolipidemic population without evidence of established cardiovascular disease (CVD). A total of 93 males and 92 females, healthy and normolipidemic, volunteered to be submitted to a clinical examination, a blood collection, and to answer a questionnaire aimed at determining signs and symptoms of atherosclerotic disease. Their fasting plasma lipid, lipoproteins, apolipoproteins, and the cholesterol and triglyceride concentrations in HDL2 and HDL3, isolated by microultracentrifugation, were determined by enzymatic-colorimetric methods. The interpercentile intervals (2.5-97.5) for the population were established as being 5-18 mg/dL in men and 4-28 mg/dL in women for HDL2 cholesterol (HDL2chol) and 1 57 mg/dL in men and 2-61 mg/dL in women for HDL3 cholesterol (HDL3chol). HDL2 triglyceride levels (HDL2Tg) in men were 1-26 mg/dL and in women 2-28 mg/dL; moreover, the HDL3 triglyceride (HDL3Tg) intervals were established as 4-46 mg/dL for both sexes. The determination of reference ranges for lipids in HDL subfractions in populations without clinical atherosclerosis, is an useful tool for metabolic, diagnostic, and therapeutic approaches. We determined the intervals for HDL2chol, HDL3chol, HDL2Tg, and HDL3Tg. There were variations with sex and/or age for HDL2chol, HDL3chol, and HDL2Tg in the studied population. PMID- 16721835 TI - Association between the TAP2 gene codon 665 polymorphism and Graves' disease. AB - A total of 95 patients with active Graves' disease (GD) and 105 normal healthy subjects were enrolled in this study, which attempted to determine whether single site polymorphisms of the transporter associated with antigen processing 2 (TAP2) gene contribute to an individual's susceptibility to GD. Such polymorphisms were detected using polymerase chain reaction (PCR)-based restriction analysis. Associations between GD and the three site polymorphisms of the TAP2 gene at codons 379, 565, and 665 were investigated. The results of the genotype analysis revealed that the frequency of the GG homozygote's presence at codon 665 was lower, and that of the AA homozygote's presence was greater in GD patients (15.8% and 36.8%, respectively) compared to normal controls (34.3% and 16.2%, respectively; P<0.001). The OR (OD) for the risk of occurrence for the AA homozygote and AG heterozygote compared to the GG homozygote (as was the case for the GD patients) was respectively 4.941 and 2.117, with respective 95% confidence intervals (CI) of 2.303-10.598 and 1.020-4.369. The allelic analysis also demonstrated reduced G and enhanced A allele frequencies for GD patients compared to controls (respectively 39.5% vs. 59.0% [G allele], and 60.5% vs. 41.0% [A allele]; P=0.0001; OR=2.219, 95% CI: 1.449-3.395). By contrast, the differences between patient and control groups for the frequency of appearance of genotypes and allelic variants at codon 379 (P=0.522 and P=0.306, respectively) and codon 565 (P=0.199 and P=0.157, respectively) did not appear to be significant. These data reveal that the single-site polymorphism of the TAP2 gene at codon 665 may be an indicator for predicting GD development. PMID- 16721836 TI - Activity of alcohol dehydrogenase (ADH) isoenzymes and aldehyde dehydrogenase (ALDH) in the sera of patients with breast cancer. AB - Numerous experiments have shown that alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) play a significant role in the metabolism of many biological substances. Some metabolic disorders that can lead to breast carcinogenesis may be the cause of changes in ADH and ALDH activity. In previous experiments we found a changed level of class I ADH activity in breast cancer tissues. The changed level of this enzyme in cancer cells may be reflected by the enzyme's activity in the serum. Therefore, in this study we measured the activity of ADH isoenzymes and ALDH in the sera of patients with breast cancer. Serum samples were taken for routine biochemical investigation from 45 women with breast cancer before treatment. Among all tested classes of ADH isoenzymes, only class I had higher activity in the serum of patients with breast cancer in stage IV. The total ADH and ALDH activities were not significantly higher in patients with breast cancer than in healthy controls. The changes in activity, especially in class I ADH, appear to be caused by isoenzymes being released from the organ damaged by metastatic disease. PMID- 16721837 TI - Evaluation of two automated systems for detection of bacteriuria. AB - The Utiscreen-CORAL Biomedical system and ROBOBACT system were tested against conventional uroculture in blood agar and MacConkey agar as a reference method to determine the bacteriuria from 400 samples. For the Utiscreen-CORAL Biomedical system, a sensitivity of 92.5% was obtained. However, by the ROBOBACT system, the sensitivity was 69.9%. PMID- 16721838 TI - PACAP 38 is involved in the non-adrenergic non-cholinergic inhibitory neurotransmission in the pig urinary bladder neck. AB - AIMS: To investigate the role played by pituitary adenylate cyclase activating polypeptide 38 (PACAP 38) in the non-adrenergic non-cholinergic (NANC) neurotransmission of the pig urinary bladder neck. METHODS: Urothelium-denuded bladder neck strips were dissected and mounted in organ baths containing a physiological saline solution (PSS) at 37 degrees C and gassed with 5% CO(2) and 95% O(2), for isometric force recording. The relaxations to transmural nerve stimulation (EFS) or PACAP 38 were performed on strips precontracted with 1 microM phenylephrine (PhE). EFS experiments were carried out in the absence and the presence of guanethidine (10 microM), atropine (0.1 microM), and N(G)-nitro-L arginine (L-NOARG, 100 microM), to block noradrenergic neurotransmission, muscarinic receptors, and nitric oxide (NO) synthase, respectively. RESULTS: EFS (2-16 Hz, 1 ms duration, 20 sec trains, 75 mA current output) evoked frequency dependent relaxations which were reduced by the VIP/PACAP receptor antagonist PACAP (6-38) (3 microM), and by the neurotoxin of the capsaicin-sensitive primary afferents capsaicin (10 microM), and abolished by the neuronal voltage-activated Na(+) channel blocker tetrodotoxin (TTX, 1 microM). The vasoactive intestinal peptide (VIP) receptor antagonist [Lys(1), Pro(2,5), Arg(3,4), Tyr(6)]-VIP (3 microM) failed to modify the EFS-induced relaxations. PACAP 38 (1 nM-1 microM) induced concentration-dependent relaxations which were reduced by PACAP (6-38), TTX and by the neuronal voltage-gated Ca(2+) channel inhibitor omega-conotoxin GVIA (omega-CgTX, 1 microM). CONCLUSIONS: The results suggest that PACAP 38, mainly released from capsaicin-sensitive primary afferents, is involved in the NANC inhibitory neurotransmission of the pig urinary bladder neck, producing relaxation through neuronal and muscle VIP/PACAP receptor activation. PMID- 16721839 TI - Ana and the Internet: a review of pro-anorexia websites. AB - OBJECTIVE: The purpose of this article is to describe the content of pro-anorexia websites, both qualitatively and quantitatively. METHOD: An Internet search protocol was developed to identify pro-anorexia websites. A grounded theory approach was used to generate themes from Internet-based information. Basic descriptive analysis was employed to report on key website characteristics. RESULTS: Twenty pro-anorexia websites met inclusion criteria. Saturation of themes was achieved after review of 12 websites. Key website characteristics included purpose of website (75%), information about webmaster (67%), website disclaimers (58%), and information on "tips and tricks" (67%). Religious metaphors, lifestyle descriptions, and "thinspiration" (inspirational photo galleries and quotes that aim to serve as motivators for weight loss) were frequently present. A total of 10 themes were generated. The most prevalent themes included control, success, and perfection. CONCLUSION: Health-care providers and caregivers should be aware of pro-anorexia websites and their content, as these websites contain information that promote and support anorexia nervosa. PMID- 16721840 TI - Does the frequency of anxiety and depressive disorders differ between diagnostic subtypes of anorexia nervosa and bulimia? AB - OBJECTIVE: The objective of the present work is to determine whether the prevalence of depressive and anxiety disorders varies in subgroups of eating disorders (ED) according to age, ED duration, mode of care provision, and body mass index (BMI). METHOD: Using the Mini International Neuropsychiatric Interview (MINI), the frequency of anxiety and depressive disorders was evaluated in 271 ED participants. Their prevalence was compared in subgroups of anorexics (AN-R and AN-BN) and bulimics (BN), both before and after controlling for potential confounding variables. RESULTS: Current or lifetime comorbidity of anxiety and depressive disorders did not differ between AN-R and AN-BN groups. Social phobia, panic disorders, and obsessive-compulsive disorder (OCD) were significantly more frequent in AN-BN and AN-R groups. Panic disorder was more frequent in the BN group. CONCLUSION: Several confounding factors, in particular those identified in the present study, may explain previous conflicting results on the frequency of anxiety and depressive disorders in ED. Nevertheless, the study confirmed that OCD is more frequent in AN, even after controlling for confounding factors. PMID- 16721841 TI - Analysis of prognostic factors regarding the outcome after a transurethral resection for symptomatic benign prostatic enlargement. AB - AIMS: To identify preoperative predictive variables regarding treatment outcomes following transurethral resection of the prostate (TURP) of patients with symptomatic benign prostatic enlargement (BPE). METHODS: A retrospective study was conducted in 384 patients of 1,397 men who had undergone TURP for symptomatic BPE. All the patients had completed the evaluation of International Prostate Symptom Score (I-PSS), and quality of life (QOL) index, and had undergone full urodynamics before the surgery. Outcomes were assessed at 12 months after surgery. The association between baseline variables and the improvement in outcome variables was statistically analyzed. RESULTS: Preoperative urodynamic abnormalities included bladder outlet obstruction (BOO) in 315 (82.0%), detrusor underactivity in 91 (23.7%), and detrusor overactivity (DO) in 160 (41.7%). Multivariate analysis showed that the presence of DO and a higher degree of BOO were both associated with postoperative improvement both in I-PSS and the QOL. The initial level of storage symptoms correlated with an improvement in the QOL. CONCLUSIONS: The association between those variables obtained by preoperatively performed urodynamic analysis and the degree of improvement in lower urinary tract symptom and QOL following TURP was statistically elucidated in a large number of patients with symptomatic BPE. A higher degree of baseline BOO positively predicts the postoperative improvement in I-PSS and QOL, while the baseline DO negatively predict it. PMID- 16721842 TI - Dopaminergic mechanisms controlling urethral function in rats. AB - AIMS: To investigate the role of dopamine receptor subtypes in the control of urethral activity. METHODS: Simultaneous recordings of intravesical and urethral perfusion pressure (UPP) were performed in rats under urethane anesthesia. Changes in coordinated activity of the bladder and urethral sphincter were examined following intravenous (i.v.), intrathecal (i.t.), or intracerebroventricular (i.c.v.) administration of dopamine D1- and D2-like receptor agonists (SKF38393 and quinpirole, respectively) and antagonists (SCH23390 and remoxipride, respectively). RESULTS: Quinpirole (0.03, 0.1, and 0.3 mg/kg i.v.) dose-dependently decreased baseline urethral pressure to 45.33 +/- 5.8, 33.7 +/- 3.3 (P < 0.05, n = 6), and 27.7 +/- 3.3 cm H(2)O (P < 0.05, n = 5) from the control value (46.0 +/- 4.0 cm H(2)O), respectively. i.c.v. injection of quinpirole (1 microg) decreased baseline urethral pressure to 33.6 +/- 5.0 cm H(2)O (P < 0.05, n = 4) from the control value (51.4 +/- 4.9 cm H(2)O) in contrast to the insignificant effects of i.t. administration of the drug (3 microg). The decrement of baseline pressure induced by quinpirole (0.1 mg/kg i.v.) was suppressed by alpha-bungarotoxin (BGT), a neuromuscular blocking agent. SCH23390 (1 and 3 mg/kg, i.v.) dose-dependently decreased the frequency of high frequency oscillation (HFO) of the urethral sphincter. SKF38393 or remoxipride did not have significant effects on any parameters of bladder and urethral activity. CONCLUSIONS: These results indicate that activation of D2-like dopamine receptors at a supraspinal site can suppress activity of the striated muscle urethral sphincter. Thus, decreased urethral resistance induced by D2 dopamine receptor activation might aggravate urge incontinence symptoms often seen in patients with Parkinson's disease (PD). PMID- 16721843 TI - The effect of attentional bias toward shape- and weight-related information on body dissatisfaction. AB - OBJECTIVE: The objective of this study was to investigate the causal role of selective attention to shape/weight-related information in terms of intensifying body dissatisfaction. METHOD: The participants were 70 female first-year psychology students aged 17-28 years. An attentional probe task was used to induce attention toward either negative shape/weight-related words, neutral words, or negatively valenced emotion words. Thereafter vulnerability toward the development of body dissatisfaction was assessed in the three groups after being exposed to a body image challenge. RESULTS: The induction of an attentional bias toward shape/weight-related information resulted in higher body dissatisfaction compared with both control groups. CONCLUSION: The results support the notion that an attentional bias toward shape/weight-related information plays a causal role in body dissatisfaction, suggesting that such biases may prove to be a useful target in interventions designed to improve body image. PMID- 16721844 TI - Multicenter quality control of the detection of HIV-1 genome in semen before medically assisted procreation. AB - Couples in whom the man is HIV-1-positive may use medically assisted procreation in order to conceive a child without contaminating the female partner. But, before medically assisted procreation, the semen has to be processed to exclude HIV and tested for HIV nucleic acid before and after processing. The performance was evaluated of the technical protocols used to detect and quantify HIV-1 in 11 centers providing medically assisted procreation for couples with HIV-1 infected men by testing panels of seminal plasma and cells containing HIV-1 RNA and/or DNA. The performance of these tests varied due to the different assays used. False positive results were obtained in 14-19% of cases. The sensitivity for RNA detection in seminal plasma was 500-1,000 RNA copies/ml, over 500 RNA copies/10(6) cells in semen cells, and for DNA detection in semen cells 50-500 DNA copies/10(6) cells. The use of silica-based extraction seemed to increase the assay performance, whereas the use of internal controls to detect PCR inhibitor did not. This first quality control highlights the need for technical improvements of the assays to detect and quantify HIV in semen fractions and for regular evaluation of their performance. PMID- 16721845 TI - Comparative performance of the Amplicor HIV-1 Monitor Assay versus NucliSens EasyQ in HIV subtype C-infected patients. AB - In facing global programs for treating HIV-infected patients in the developing countries, there is a real need for viral load assays that are accurate for the local subtypes. The present study was designed to evaluate viral load measurements using the newer version of the NASBA assay in subtype C-infected patients. The performances of this new version, a real-time nucleic acid sequence based amplification HIV-1 assay (NucliSens EasyQ), were compared to Amplicor HIV 1 Monitor Assay version 1.5 in 79 samples of subtype C-infected patients originating from Ethiopia. Twenty HIV-1 subtype B-infected patients served as a control group. Blood samples from patients in both groups were tested by both assays. The results were compared by a paired, two-tailed Student's t-test. The disparity between the results of the two viral load assays was highly significant in subtype C samples (P = 0.005), such that in the vast majority, higher values of viral load were obtained by the Amplicor assay. However, no differences between the two assays were found in subtype B samples (P = 0.77). CD4 measurements were available for 78 samples of subtype C-infected patients. Of these, a CD4-to-viral load discrepancy (CD4 20 pp65 + positive cells, median CMV DNA value: 3.8 log(10) copies/500,000 PBLs) than in not-treated ones (2.9 logs). According to pp65 levels of 0, 1-10, 11-20, 21-50, 51-100, and >100 positive cells/200,000 PBLs, median CMV DNA by real-time PCR was 2.6, 3.0, 3.6, 4.0, 4.2, and 4.8 logs, respectively, (CMV DNA levels by COBAS AMPLICOR: 2.8, 2.9, 3.8, 3.7, 3.9, and 4.0 logs). For samples with >20 pp65 + cells, real-time PCR gave significantly higher values than in groups with <20 pp65 + cells, whereas the COBAS AMPLICOR results showed a slower progression rate. Dilutions of CMV AD169 strain were used to probe real-time PCR reproducibility (between and intra-assay variability <2%) and sensitivity (100% detection rate at 10 copies/reaction, 28.5% with end-point PCR). In conclusion, real-time PCR significantly improves the study of CMV DNA dynamics due to a more reliable quantitation of CMV DNA for moderate and high DNA level compared to end-point PCR with better sensitivity and specificity. Real time PCR provides more precise information for evaluating infection progress and assessing antiviral response, simplifying and accelerating the process of producing a reliable quantitation of CMV DNA for clinical purposes. PMID- 16721849 TI - Genetic variability of human coronavirus OC43-, 229E-, and NL63-like strains and their association with lower respiratory tract infections of hospitalized infants and immunocompromised patients. AB - In the winter-spring seasons 2003-2004 and 2004-2005, 47 (5.7%) patients with acute respiratory infection associated with human coronavirus (hCoV) 229E-, NL63 , and OC43-like strains were identified among 823 (597 immunocompetent and 226 immunocompromised) patients admitted to hospital with acute respiratory syndromes. Viral infections were diagnosed by either immunological (monoclonal antibodies) or molecular (RT-PCR) methods. Each of two sets of primer pairs developed for detection of all CoVs (panCoV) failed to detect 15 of the 53 (28.3%) hCoV strains identified. On the other hand, all hCoV strains could be detected by using type-specific primers targeting genes 1ab and N. The HuH-7 cell line was found to be susceptible to isolation and identification of OC43- and 229E-like strains. Overall, hCoV infection was caused by OC43-like, 229E-like, and NL63-like strains in 25 (53.2%), 10 (21.3%), and 9 (19.1%) patients, respectively. In addition, three patients (6.4%) were infected by untypeable hCoV strains. NL63-like strains were not found to circulate in 2003-2004, and 229E like strains did not circulate in 2004-2005, while OC43-like strains were detected in both seasons. The monthly distribution reached a peak during January through March. Lower predominated over upper respiratory tract infections in each age group. In addition, hCoV infections interested only immunocompetent infants and young children during the first year of life, while all adults were immunocompromised patients. Coinfections of hCoVs and other respiratory viruses (mostly interesting the first year of life) were observed in 14 of the 47 (29.8%) patients and were associated with severe respiratory syndromes more frequently than hCoV single infections (P = 0.002). In conclusion, the use of multiple primer sets targeting different genes is recommended for diagnosis of all types of hCoV infection. In addition, the detection of still untypeable hCoV strains suggests that the number of hCoVs involved in human pathology might further increase. Finally, hCoVs should be screened routinely for in both infants and immunocompromised patients with acute respiratory infection. PMID- 16721850 TI - Changing distribution of norovirus genotypes and genetic analysis of recombinant GIIb among infants and children with diarrhea in Japan. AB - A total of 402 fecal specimens collected during July 2003-June 2004 from infants and children with acute gastroenteritis, encompassing five localities (Maizuru, Tokyo, Sapporo, Saga, and Osaka) of Japan, were tested for the presence of norovirus by RT-PCR. It was found that 58 (14.4%) fecal specimens were positive for norovirus. Norovirus infection was detected throughout the year with the highest prevalence in December. Norovirus GII was the most predominant genogroup (98.3%; 57 of 58). The genotypes detected in this study were GI/4, GII/2, GII/3, GII/4, and GII/6. Of these, NoV GII/3 (known as the Arg320 virus cluster) was the most predominant genotype (43.9%), followed by NoV GII/4 (the Lordsdale virus cluster; 35.1%) and others. Two norovirus strains clustered with a "new variant designated GIIb" and a "new variant of GII/4" were found circulating in Japan for the first time. It was interesting to note that NoV GIIb and NoV GII/3 appeared to be the recombinant strains and the recombination site was demonstrated at the overlap of ORF1 and ORF2. The majority (96%) of the dominant norovirus strains were identified as the recombination of GII/3 capsid and GII/12 polymerase. The recombination in the NoV GIIb capsid gene at the breakpoint located at P1 domain was also identified. Obviously, NoV GIIb isolate in Japan had double recombination. This is the first report demonstrating the existence of different "new variants" co-circulating in Japanese infants and children with acute gastroenteritis. PMID- 16721851 TI - Re: request from the International Advisory Committee to DDBJ/EMBL/GenBank. PMID- 16721852 TI - Comparative analysis of purified Pacific and Caribbean ciguatoxin congeners and related marine toxins using a modified ELISA technique. AB - The monoclonal antibody to ciguatoxin (CTX) produced from a hybridoma cell line was assayed for the detection of four congeners of CTX: Pacific ciguatoxin-1 (P CTX-1), Pacific ciguatoxin-2 (P-CTX-2), Pacific ciguatoxin-3 (P-CTX-3), and Caribbean ciguatoxin-1 (C-CTX-1) and related marine toxins, including domoic acid, palytoxin, and okadaic acid, using a modified enzyme-linked immunosorbent assay (ELISA). Lower detection limits were assessed and linearity was statistically established (P<0.05) for P-CTX-1, P-CTX-2, and P-CTX-3 and C-CTX-1 at concentrations ranging from 0 to 5.00 ng, while the other marine toxins showed statistically insignificant cross-reactivities at similar concentrations. Thus, the monoclonal antibody to CTX is able to specifically detect various CTX congeners at levels comparable to those naturally occurring in ciguatoxic fish. PMID- 16721853 TI - Characterization of truncated hantavirus nucleocapsid proteins and their application for serotyping. AB - Hemorrhagic fever with renal syndrome (HFRS) is a fulminant infectious disease characterized by fever, hemorrhage, renal impairment, and thrombocytopenia. Hantaviruses associated with this belong to different serotypes: Hantaan (HTN), Seoul (SEO), Dobrava/Belgrade (DOB), and Puumala (PUU). The first two, HTN and SEO, are endemic in China. To investigate the epidemiology of HFRS and virus transmission in China, we constructed prokaryotic plasmids encoding truncated recombinant HTN and SEO nucleocapsid proteins (NPs), which lacked 154 amino acid (aa), 99 aa, or 49 aa in the N-terminal region, respectively. After expression, the truncated rNPs were tested as serotyping antigens, particularly for use in the enzyme-linked immunosorbent assay (ELISA). In addition, 68 acute and 52 convalescent sera were collected from HFRS patients from Harbin, Lantian, and Kaifeng regions in China in 2004, which had hantavirus specific antibodies by IFA. A neutralization test was used to differentiate these, which showed that 73 were due to HTN infection, 33 to SEO infection, and 14 undetermined. By ELISA, the truncated rNPs, that lacked 99 (rNP100) or 49 (rNP50) N-terminal amino acids of the NPs of HTN and SEO, were able to differentiate HTNV and SEOV-specific immune sera, but the rNP155 could not. Particularly, the ELISAs based on the rNP50s had a result comparable to PRNT. Thus, the rNP50 is recommended as efficient serotyping antigen for hantavirus infection diagnosis by ELISA. PMID- 16721854 TI - Protection against influenza virus infection by intranasal vaccine with surf clam microparticles (SMP) as an adjuvant. AB - A safe and effective adjuvant is necessary to enhance mucosal immune responses for the development of an inactivated intranasal influenza vaccine. The present study demonstrated the effectiveness of surf clam microparticles (SMP) derived from natural surf clams as an adjuvant for an intranasal influenza vaccine. The adjuvant effect of SMP was examined when co-administered intranasally with inactivated A/PR8 (H1N1) influenza virus hemagglutinin vaccine in BALB/c mice. Administration of the vaccine with SMP induced a high anti-PR8 haemagglutinin (HA)-specific immunoglobulin A (IgA) response in the nasal wash and immunoglobulin G (IgG) response in the serum, resulting in protection against both nasal-restricted infection and lethal lung infection by A/PR8 virus. In addition, administration of SMP with A/Yamagata (H1N1), A/Beijing (H1N1), or A/Guizhou (H3N2) vaccine conferred complete protection against A/PR8 virus challenge in the nasal infection model, suggesting that SMP adjuvanted vaccine can confer cross-protection against variant influenza viruses. The use of SMP is suggested as a new safe and effective mucosal adjuvant for nasal vaccination against influenza virus infection. PMID- 16721855 TI - Role of nitric oxide during rotavirus infection. AB - The pathophysiological mechanisms behind rotavirus-induced diarrhoea still remain incomplete. Current views suggest that the non-structural protein 4 (NSP4) of rotavirus and the enteric nervous system (ENS) participate in water secretion and diarrhoea. In the present work the role of nitric oxide (NO) in rotavirus infection and disease has been studied in vitro, mice and humans. Incubation of human intestinal epithelial cells (HT-29) with purified NSP4 but not with infectious virus produced NO2/NO3 accumulation in the incubation media. The NSP4 induced release of NO metabolites occurred within the first minutes after the addition of the toxin. Mice infected with murine rotavirus (strain EDIM) accumulated NO2/NO3 in the urine at the onset for diarrhoea. Following rotavirus infection, inducible nitric oxide synthetase (iNOS) mRNA was upregulated in ileum, but not in duodenum or jejunum of newborn pups within 5 days post infection. A prospective clinical study including 46 children with acute rotavirus infection and age-matched controls concluded that rotavirus infection stimulates NO production during the course of the disease (P < 0.001). These observations identify NO as an important mediator of host responses during rotavirus infection. PMID- 16721856 TI - Long-term follow-up of hepatitis B carrier children treated with interferon and prednisolone. AB - The long-term outcome of treatment with Interferon Alpha 2B with and without Prednisolone priming in children infected perinatally with hepatitis B was reviewed. The group studied included 48 children (aged 2-16 years), who were HBe antigen and hepatitis B DNA positive between 1991 and 1993. Twenty children were randomized to a therapeutic trial at that time, and received Prednisolone in reducing doses for 6 weeks and Interferon for 16 weeks while 22 children were monitored without treatment for 12 months. Fourteen of the untreated group and 6 additional children later received treatment with Interferon alone (n = 20). Eight children for whom treatment was declined were followed long term. Median follow-up was 7.5 years (range 1.5-10.6). There was no significant effect of Interferon therapy on seroconversion with or without Prednisolone at 12 months post-treatment compared to untreated children. On longer term follow-up, the 5 year HBeAg to anti-HBe seroconversion percentages, estimated from Kaplan-Meier curves, were 54% for Prednisolone plus Interferon, 22% for Interferon alone, and 12% for untreated children. The median time to seroconversion was 3.9 years (range 0.4-8.2) and was shortest in those treated with Prednisolone plus Interferon. Children who had elevated hepatic transaminase enzymes prior to treatment or during Prednisolone priming had a better response. In contrast to many European studies, no child cleared HBsAg and produced anti-HBs. Treatment with Prednisolone priming and Interferon, improved both the time and rate of seroconversion compared to no treatment or Interferon alone, suggesting that this combination of drugs might have an immunomodulatory effect. PMID- 16721857 TI - Emergence and takeover of precore-stop mutant prior to exacerbation of e antigen negative chronic hepatitis B after withdrawal of lamivudine therapy. AB - Although hepatitis B virus (HBV) precore-stop mutant (nonsense mutation at codon 28) is usually isolated from patients with hepatitis B e antigen (HBeAg)-negative hepatitis, it is unclear whether the mutant is the cause of the hepatitis flare or it is a consequence of necroinflammation. Using the amplification-created restriction site method, the ratios of precore-stop mutant were measured in serial serum samples obtained from three patients experiencing HBeAg-negative hepatitis after withdrawal of lamivudine therapy. In all three patients, the precore-stop mutant emerged and took over the viral population at least 2 months prior to the hepatitis exacerbations. After the onset of exacerbation, transient elevation of serum HBV-DNA was observed, which was followed by progressive decrease of the ratio of precore-stop mutant as well as normalization of serum transaminases. The present data indicate that at least in some patients, the precore-stop mutant is the cause but not the consequence of the flare of HBeAg negative hepatitis. PMID- 16721858 TI - Alanine transaminase levels in the year before pregnancy predict the risk of hepatitis C virus vertical transmission. AB - Vertical transmission is the most common route of hepatitis C virus (HCV) infection in children. Transmission risk factors have been described, but most risk factors can only be evaluated using expensive laboratory exams. The aim of the present study was to evaluate whether maternal alanine transaminase (ALT) levels before pregnancy correlate with HCV vertical transmission. Seventy-four transmitting and 403 nontransmitting mothers were evaluated. All mothers enrolled had two ALT determinations in the last year before pregnancy, at least 6 months apart. Mothers were divided into two groups: mothers with persistently normal serum ALT levels and mothers with abnormal ALT levels. In the second group both mothers with constantly raised or with fluctuating ALT levels (one normal and one raised determination) were included. ALT was defined as raised if higher than twice the upper limit of normal. Abnormal ALT levels were found in 39/74 (52.7%) HCV transmitting mothers and in 146/403 (32.6%) nontransmitting mothers (P = 0.008; relative risk 1.96; 95% confidence limits 1.19-3.23). The risk of transmission from mothers with constantly raised ALT levels was more evident than that from mothers with fluctuating ALT levels. Increased ALT levels may reflect a more severe liver disease and a higher viral load, factors known to be associated with vertical transmission. ALT determination, a simple, widely available and inexpensive test, may help in identifying mothers with an increased risk of HCV vertical transmission. PMID- 16721859 TI - CMV DNA detection in dried blood spots for diagnosing congenital CMV infection in Japan. AB - Human cytomegalovirus (CMV) is a leading congenital infectious agent in developed countries. In the past, the incidence of congenital infection has been rather low in Japan because a high seroprevalence of CMV present in young women. However, this seroprevalence has been decreasing in recent years, so that the incidence of congenital CMV infection in Japanese neonates may increase and approach the level seen in other developed countries. The method was used for detecting CMV DNA reported by Barbi et al. [Barbi et al. (1996): Clin Diagn Virol 6:27-32] using a dried blood spot on filter paper, to diagnose congenital CMV infection in Japanese neonates. This method is effective and less laborious than virus isolation both for epidemiological studies and for identifying asymptomatic infected babies. Japanese neonates (1,176) were examined; two of who were asymptomatic were found to be infected. PMID- 16721860 TI - High human herpesvirus 8 seroprevalence in populations from Western Balkan countries. AB - Patterns of endemicity of human herpesvirus 8 (HHV8) are still undefined in some European populations, such as those from Western Balkan countries. Serum samples from 605 human immunodeficiency virus-seronegative subjects (299 Albanians and 306 Kosovars) were tested for the presence of HHV8 antibodies to a capsid-related open reading frame (ORF65)-encoded protein and a latency-associated nuclear antigen (LANA) to determine HHV8 seroprevalence in populations from Albania and from the Kosovo region of former Yugoslavia. Levels of co- circulation with hepatitis A (HAV) and hepatitis B (HBV) viruses were also determined. HHV8 antibodies to at least one of the two antigens were detected in 28.8% of Albanians and 18% of Kosovars. The seroprevalence of HHV8 was found to be 25.0 and 16.8% in Albanian and Kosovar children (1>2. The DNA-binding and -photocleavage characteristics of the three cationic water-soluble chlorin e(6) derivatives are influenced by aspects of their molecular structure, such as the kind, number, and position of the cationic substituents. PMID- 16721874 TI - Isolation and identification of mixed linked beta -glucan degrading bacteria in the intestine of broiler chickens and partial characterization of respective 1,3 1,4-beta -glucanase activities. AB - Media with 1,3-1,4-beta -glucans as selective markers were used for isolation of non-starch-polysaccharide (NSP) degrading bacteria from the intestinal tract of broiler chicken. Formerly unknown 1,3-1,4-beta endoglucanase activities in various bacterial species were identified in this study. E. faecium , Streptococcus , Bacteroides and Clostridium strains seem to be responsible for degradation of mixed linked beta -glucans in the small intestine and in the hind gut of chickens. Strict anaerobic bacteria (Bacteroides ovatus , B. uniformis , presumably B. capillosus and Clostridium perfringens ) as well as an unidentified bacterium with 98% 16S rDNA homology to an uncultered chicken cecum bacterium were isolated. Additionally, Streptococcus bovis with 1,3-1,4-beta -endoglucanase activity was also detected. Different 1,3-1,4-beta -endoglucanase activity profiles were observed in SDS/PAGE zymograms. PMID- 16721875 TI - Characterization of bacteriocin HV219, produced by Lactococcus lactis subsp. lactis HV219 isolated from human vaginal secretions. AB - Bacteriocin HV219, produced by Lactococcus lactis subsp. lactis HV219, is active against Gram-positive and Gram-negative bacteria. Activity was lost when treated with proteolytic enzymes, SDS, Triton X-114 and Triton X-100, but not at pH 2.0 to 10.0 or after 20 min at 121 degrees C. Growth in the presence of yeast extract as sole nitrogen source yielded 3200 AU/ml. No bacHV219 activity was recorded in MRS broth with maltose, mannose, lactose or sucrose as sole carbohydrate, but fructose yielded 1600 AU/ml. K(2)HPO(4) at 10.0 g/l yielded 3200 AU/ml. Addition of 1.0 mg/l cyanocobalamin, l-ascorbic acid and thiamine to MRS broth yielded 3200 AU/ml, 1600 AU/ml and 1600 AU/ml, respectively. The mode of activity is bacteriolytic, as confirmed by atomic force microscopy. PMID- 16721876 TI - A new p-terphenyl derivative from fruiting bodies of the basidiomycete Sarcodon laevigatum. AB - A new metabolite with p -terphenyl core, named sarcodan (1), together with three known p -terphenyl metabolites (3, 4, 5) represented as Fig. 1, was isolated from the fruiting bodies of the basidiomycete Sarcodon laevigatum . The structures of these compounds were elucidated by spectroscopic and chemical methods. PMID- 16721877 TI - Practical thin layer chromatography techniques for diaminopimelic acid and whole cell sugar analyses in the classification of environmental actinomycetes. AB - For the determination of diaminopimelic acid (DAP) stereoisomers in whole cell hydrolysates as chemotaxonomic markers of actinomycetes, two new pyridine-free solvent systems for TLC on cellulose sheets have been introduced: methanol/0.05 m potassium hydrogenphthalate buffer pH 4 2:1 (v/v) and methanol/0.12 M dimethylaminopyridine (DMAP) in H(2)O pH 6 2:1 (v/v). The commercial (Sigma) DAP standard can be separated by repeated TLC into its three stereoisomers. Addition of a single stereoisomer to the samples supported the detection (presence or absence) of the relevant stereoisomer by HPLC. In the study on whole cell sugars, TLC both on cellulose and on silica gel revealed to be successful in the separation of the components in modified pyridine-free solvent systems. The procedures of DAP and sugar analyses are summarized in two flow-charts. PMID- 16721878 TI - Plant growth promotion and induction of resistance in Camellia sinensis by Bacillus megaterium. AB - Bacillus megaterium DE BARY TRS-4 was isolated from tea rhizosphere and tested for its ability to promote growth and cause disease reduction in tea plants. In vivo studies revealed the ability of this bacterium to promote growth of tea plants very significantly. Brown root rot disease, caused by Fomes lamaoensis was markedly reduced by application of the bacterium to the soil. Population of F. lamaoensis in soil before and after application of B. megaterium, as determined by ELISA and dot-blot using PAb raised against the pathogen, was shown to be greatly reduced in presence of the bacterium. Biochemical changes induced in tea plants were also examined. Root colonization by B. megaterium and subsequent inoculation with F. lamaoensis also led to an increase in polyphenolics, as well as in defense related enzymes-peroxidase, chitinase, beta -1,3-glucanase and phenyl alanine ammonia lyase. Determination of mechanism of action of this bacterium revealed it to be able to solubilize phosphate, produce IAA, siderophore and antifungal metabolite. The plant growth promotion and reduction of disease intensity have been shown to be due to a combination of several mechanisms. PMID- 16721879 TI - A mathematical model of Saccharomyces cerevisiae growth in response to cadmium toxicity. AB - Microbial growth can be described using models derived by differential equations, but available mathematical models have yet to adequately describe lag phase related cell growth or cell mortality in response to chemical toxicity. Lag phase cell behavior, however, dictates the onset of exponential growth and the number of actively growing cells available to initiate exponential growth, important factors in the success of remediation efforts. In this study, a five-parameter polynomial ratio (PR) model was used to characterize the growth, from lag through stationary phase, of the yeast Saccharomyces cerevisiae in response to cadmium toxicity. The PR model used in this study has the advantages over standard mathematical models in the ability to represent the initial cell mortality observed when S. cerevisiae is exposed to increasing cadmium levels, up to 12 mg/l Cd, as well as following cell recovery and growth to stationary levels. PMID- 16721880 TI - Evaluation of antibacterial activity of extracts of five species of wood colonizing fungi. AB - Screening new organisms for antibacterial activity and searching for new antibacterial drugs is important due to the constant generation of new antibiotic resistant strains of pathogenic bacteria. An E. coli broth microdilution test was used to evaluate the results of the Vibrio fischeri bioluminescence test in five of the most antibacterially active species of wood-colonizing fungi. Serpula lacrymans was found to be a potential source of thermostable antibiotic(s) and the Vibrio fischeri bioluminescence test was confirmed to be a useful method for screening for antibacterial activity. PMID- 16721881 TI - Isolation, identification and seasonal distribution of soilborne fungi in tea growing areas of Iyidere-Ikizdere vicinity (Rize-Turkey). AB - This study involves the isolation and identification of soil borne fungi from different altitudes of Iyidere-Ikizdere vicinity in Rize, Turkey. The qualitative and quantitative fungal composition of the soil of Iyidere-Ikizdere vicinity was surveyed seasonally during a year (between summer 2003 and spring 2004). The soil samples were collected from 10 different locations. Some physical and chemical properties of the soil samples were studied. Soil samples were observed to have the proper pH levels (pH 4.00-6.75) for the growth of microfungi. By using a soil dilution technique, the mycological analysis of 40 soil samples were studied at 25 cm depth. The number of microfungi ranged from 2,000-160,000 CFU g(-1). 249 different microfungi belonging to 15 genera were isolated. 45 of the isolates were Mycelia Sterilia. Penicillium, Aspergillus, Trichoderma and Fusarium were the most abundant genera (26.8%, 14.8%, 9.6% and 8.4%, respectively). Penicillium phialosporum, Penicillium trzebinskianum and Gliocladium virens were reported for the first time in Turkey. PMID- 16721882 TI - Screening and selection of most potent diazotrophic cyanobacterial isolate exhibiting natural tolerance to rice field herbicides for exploitation as biofertilizer. AB - Periodic applications of heavy dosages of herbicides in modern rice-agriculture are a necessary evil for obtaining high crop productivity. Such herbicides are not only detrimental to weeds but biofertilizer strains of diazotrophic cyanobacteria also. It is therefore, essential to screen and select such biofertilizer strains of diazotrophic cyanobacteria exhibiting natural tolerance to common rice-field herbicides that can be further improved by mutational techniques to make biofertilizer technology a viable one. Therefore, efforts have been made to screen five dominant diazotrophic cyanobacterial forms e.g. filamentous heterocystous Nostoc punctiforme , Nostoc calcicola , Anabaena variabilis and unicellular Gloeocapsa sp. and Aphanocapsa sp. along with standard laboratory strain Nostoc muscorum ISU against increasing concentrations (0-100 mg l(-1) of four commercial grade common rice-field herbicides i.e. Arozin, Butachlor, Alachlor and 2,4-D under diazotrophic growth conditions. The lethal and IGC(50) concentrations for all four herbicides tested were found highest for A. variabilis as compared to other test cyanobacteria. The lowest reduction in chlorophyll a content, photosynthetic oxygen evolution, and N(2)-fixation was found in A. variabilis as compared to other rice field isolates and standard laboratory strain N. muscorum ISU. On the basis of prolong survival potential and lowest reductions in vital metabolic activities tested at IGC(50) concentration of four herbicides, it is concluded that A. variabilis is the most potent and promising cyanobacterial isolate as compared with other forms. This could be further improved by mutational techniques for exploitation as most potential and viable biofertilizer strain. PMID- 16721884 TI - Triplet MLCT photosensitization of the ring-closing reaction of diarylethenes by design and synthesis of a photochromic rhenium(I) complex of a diarylethene containing 1,10-phenanthroline ligand. AB - Synthesis of the diarylethene-containing ligand L1 based on Suzuki cross-coupling reaction between thienyl boronic acid and the dibromophenanthroline ligand is reported. On coordination to the rhenium(I) tricarbonyl complex system, the photochromism of L1 could be photosensitized and consequently extended from intraligand excitation at lambda< or =340 nm in the free ligand to metal-to ligand charge-transfer (MLCT) excitation at lambda< or =480 nm in the complex. The photochromic reactions were studied by (1)H NMR, UV/Vis, and steady-state emission spectroscopy. Photosensitization was further probed by ultrafast transient absorption and time-resolved emission spectroscopy. The results provided direct evidence that the formation of the closed form by the MLCT sensitized photochromic process was derived from the (3)MLCT excited state. This supports the photosensitization mechanism, which involves an intramolecular energy-transfer process from the (3)MLCT to the (3)IL(L1) state that initiated the ring-closure reaction. The photophysical and electrochemical properties of the complex were also investigated. PMID- 16721885 TI - The reaction mechanism of a complex intercalation system: in situ X-ray diffraction studies of the chemical and electrochemical lithium intercalation in Cr4TiSe8. AB - The intercalation reaction between Cr(4)TiSe(8) and Li was investigated from a kinetic and an electrochemical perspective. The structural phase transition from monoclinic to trigonal symmetry was probed by in situ energy-dispersive X-ray diffraction (in situ EDXRD) for chemical intercalation with butyllithium (BuLi). A change in the kinetic mechanism was detected for the reaction at room temperature; this was interpreted in terms of a trend from phase boundary control to diffusion control. A single diffusion-controlled mechanism is obeyed at 60 degrees C. The electrochemical measurements and the corresponding in situ X-ray diffraction (in situ XRD) data revealed that the monoclinic host is intercalated up to the composition Li(x approximately 0.1)Cr(4)TiSe(8) before the characteristic phase transition starts. The monoclinic phase undergoes complex structural changes in the following two-phase regime. Owing to the co-existence of two phases, the cell potential is constant for 0.1150 kPa, more than 5 times higher than that of the PDMS hybrid microchip with reversible sealing. In addition, it shows an excellent heat-dissipating property and provides a user-friendly rigid interface just like a glass microchip, which facilitates manipulation of the microchip and fix tubing. As an application, PDMS sandwich microchips were tested in the capillary electrophoresis separation of fluorescein isothiocyanate-labeled amino acids. PMID- 16721902 TI - DNA fingerprinting of Vibrio cholerae and Aeromonas species by pulsed-field minigel electrophoresis. AB - DNA molecules of Vibrio cholerae and Aeromonas species were prepared by incubating immobilized cells for 4 and 2 h, respectively, with a nonenzymatic solution that contains chemical reagents only (NDSUPlus). This method gave results as reproducible as the enzymatic one that uses proteinase K, and rendered DNA molecules suitable for fingerprinting by mini-CHEF electrophoresis. As rapid DNA separations at high electric field are achieved in mini-CHEF chamber with low heat evolution, DNA restriction fragments were separated in 5 h at 10 V/cm in a single resolution window. Then, fragment separations in three resolution windows were done in 15 h. This time is shorter than the one needed by the large CHEF chamber for resolving fragments in a single resolution window. Three windows permitted to include larger numbers of restriction fragments in the calculation of isolate similarities. Both sample preparation and mini-CHEF electrophoresis may represent an alternative for performing massive epidemiological studies of V. cholerae and Aeromonas species. PMID- 16721903 TI - Dissection of mitochondrial superhaplogroup H using coding region SNPs. AB - Haplogroup H (hg H) includes about 40-50% of the West Eurasian mitochondrial DNA (mtDNA) samples investigated so far. In order to enhance discrimination within this haplogroup we selected 45 coding region SNPs that allow to ascribe samples to the main phylogenetic branches of super hg HV (that embraces hg H) and, in particular, to H sublineages with a much finer resolution than previous studies. SNP selection was carried out using the most up-to-date available literature on population and forensic genetics and extended by means of phylogenetic analysis of complete or coding region genomes (<430) and control region sequences. A meticulous inspection of the H phylogeny led us to the observation of various but uncharacterized subclades of hg H. The selected SNPs were amplified in two PCR multiplex reactions and subsequently targeted in three single-base extension multiplex reactions. A total of 2214 West Eurasian samples were screened for hg H specific loci 2706 and 7028, of which 859 fell in hg H and were further subjected to subhaplogroup typing. We observed 35 different subhaplogroups in total, 33 of which were found at frequencies below 5%. This assay can be used as a prescreening tool in forensic casework for rapid discrimination between divergent lineages (very effective for high-volume crime cases) or as discriminatory assay, when identical hg H haplotypes were obtained by control region sequencing. PMID- 16721904 TI - Micellar electrokinetic chromatography of fluorescently labeled proteins on poly(dimethylsiloxane)-based microchips. AB - MEKC of standard proteins was investigated on PDMS microfluidic devices. Standard proteins were labeled with AlexaFluor(R) 488 carboxylic acid tetrafluorophenyl ester and filtered through a size-exclusion column to remove any small peptides and unreacted label. High-efficiency MEKC separations of these standard proteins were performed using a buffer consisting of 10 mM sodium tetraborate, 25 mM SDS, and 20% v/v ACN. A separation of BSA using this buffer in a 3.0 cm long channel generated a peak with a plate height of 0.38 microm in <20 s. Additional fast separations of myoglobin, alpha-lactalbumin, lysozyme, and cytochrome c also yielded peaks with plate heights ranging from 0.54 to 0.72 microm. All proteins migrated with respect to their individual pIs. To improve the separations, we used a PDMS serpentine chip with tapered turns and a separation distance of 25 cm. The number of plates generated increased linearly with increasing separation distance on the extended separation channel chips; however, the resolution reached an asymptotic value after about 7 cm. This limited the peak capacity of the separation technique to 10-12. PMID- 16721905 TI - Solid-phase extraction and large-volume sample stacking with an electroosmotic flow pump in capillary electrophoresis for determination of methotrexate and its metabolites in human plasma. AB - This paper describes approaches for large-volume sample stacking (LVSS) with an EOF pumpin CE for the determination of methotrexate (MTX) and its metabolites in human plasma. After pretreatment of plasma through a SPE cartridge, a large sample volume was loaded by hydrodynamic injection (3 psi, 70 s) into the capillary filled with phosphate buffer (70 mM, pH 6.0) containing 0.01% polyethylene oxide. Following removal of a large plug of sample matrix from the capillary using polarity switching (-25 kV), the separation of anionic analytes was subsequently performed without changing polarity again, achieving an improvement of sensitivity of around a 100-fold. The method was applied to therapeutic drug monitoring of MTX in one acute lymphoblastic leukemia patient. This study is one of very few applications showing the feasibility of LVSS in analysis of biological samples by CE. PMID- 16721906 TI - Proteomic mapping of the hyperthermophilic and acidophilic archaeon Sulfolobus solfataricus P2. AB - A proteomic map of Sulfolobus solfataricus P2, an archaeon that grows optimally at 80 degrees C and pH 3.2, was developed using high-resolution 2-DE and peptide mass fingerprinting. A total of 867 protein spots (659 aqueous Tris-soluble spots and 208 aqueous Tris-insoluble) were mapped over IPG 3-10, 4-7, and 6-11, with second-dimensional gels made of 8-18% polyacrylamide. Three hundred and twenty four different gene products were represented by the 867 spots, with 274 gene products being identified in the Tris-soluble fractions and 100 gene products in the Tris-insoluble portion. Fifty gene products were found on gels from both fractions. Additionally, an average of 1.50 +/- 0.12 isoforms/protein was identified. This mapping study confirmed the expression of proteins involved in numerous metabolic, transport, energy production, nucleic acid replication, translation, and transcription pathways. Of particular interest, phosphoenolpyruvate carboxykinase (SSO2537) was detected even though the pathway for gluconeogenesis is unknown for this archaeon. Tris-soluble fractions contained many cytosolic proteins while Tris-insoluble fractions contained many membrane-associated proteins, including ABC transporters and an ATP synthase. This study provides an optimized 2-DE approach for investigating the biochemical pathways and post-translational modifications employed by Sulfolobus to survive in its extreme environment. PMID- 16721907 TI - Scanning copy number and gene expression on the 16p13.3-13.2 chromosomal region by the systematic multiplex polymerase chain reaction and reverse transcription polymerase chain reaction methods. AB - We developed the systematic multiplex reverse transcription-PCR (SM RT-PCR) method that is distinguishable from other multiplex RT-PCR methods by optimized PCR conditions allowing amplification of sequences that fall within a single exon of genes of similar band intensity using genomic DNA template as a calibration standard. Using an SM RT-PCR system of proto-oncogenes and tumor suppressor genes, we previously showed that the SM RT-PCR system, which was developed for cDNA expression analysis, could also be used for a more exquisite analysis of copy number changes in genomic DNA. Here we report that the SM PCR method semiquantitatively detected less than a two-fold difference in copy number. Furthermore, we also report the results of subchromosomal scanning of copy number and expression using the SM PCR and SM RT-PCR methods. We identified and characterized the novel homozygous deletion that spans over 12-plus genes on 16p13.3-13.2 in the MDA-MB-468 breast cancer cell line. PMID- 16721908 TI - Capillary electrophoresis method for determination of D-serine and its application for monitoring of serine racemase activity. AB - Serine racemase (SR) is an enzyme responsible for the biosynthesis of D-serine, the coagonist of the N-methyl-D-aspartate receptor, in the brain. Therefore, it has been suggested as a possible therapeutic target for the treatment of various neurodegenerative diseases. To develop a potent inhibitor of SR, a simple, sensitive, fast, and robust assay is needed. In this paper, a new CE method for the determination of D-serine is described. Serine enantiomers are resolved in the form of o-phthaldialdehyde (OPA)/2-mercaptoethanol (2-ME) derivatives in an alkaline BGE composed of 50 mM sodium tetraborate, pH 9.7, and containing 40 mM 2 hydroxypropyl-gamma-CD as a chiral selector. The problem of time-limited stability of OPA/2-ME derivatives has been overcome by employing in-capillary derivatization of the sample, i.e., the derivatization reaction was carried out directly in the separation capillary in the first phase of the CE run. UV absorption detection at 230 nm allowed concentration detection limit of 3 microM. Baseline resolution of D- and L-serine derivatives was achieved in less than 10 min. This fact, together with the simple sample pretreatment, allowed application of the method to medium-throughput monitoring of SR activity, such as the screening of potential SR inhibitors. A good agreement was achieved between the developed CE method and the previously established HPLC method for determination of the inhibition constant, K(i), of a new SR inhibitor, L-erythro-3 hydroxyaspartate. PMID- 16721909 TI - Global. UNAIDS: access to HIV therapy tripled in 2 years. PMID- 16721910 TI - Advocacy. Activists to lobby Congress for HIV/AIDS program funding. PMID- 16721911 TI - Court-ordered HIV testing remanded due to scant evidence. PMID- 16721912 TI - Prisons. HIV-positive inmate's retaliation charges move forward. PMID- 16721913 TI - Grievance system trips up inmate's HIV-exposure claim. PMID- 16721914 TI - HIV-testing order reversed for sexual offender. PMID- 16721916 TI - Misconduct mayhem. PMID- 16721915 TI - Thinking big. PMID- 16721917 TI - Postdoc prep. PMID- 16721918 TI - Services. HIV organizations map emergency plans for responding to disasters. PMID- 16721919 TI - Testing. Delaware proposes to test all inmates for TB, hepatitis and HIV. PMID- 16721920 TI - Treatment. HIV-positive inmate's negligence claims dismissed. PMID- 16721921 TI - ADA. Prison officials denied dismissal on HIV-bias charges. PMID- 16721922 TI - Dismissal denied UPS officials over HIV disclosure. PMID- 16721923 TI - Charges filed. Illinois, Florida suspects face HIV exposure charges. PMID- 16721924 TI - Global. HIV/AIDS issues to be tackled at church convention. PMID- 16721925 TI - New studies cool hopes for stem cell heart repairs. PMID- 16721926 TI - Rx for lower risk: a healthy dose of optimism. PMID- 16721927 TI - Atrial fibrillation raised its head 15 months ago for the first time, and medication has stabilized me. An echocardiogram at the time of diagnosis showed an ejection fraction of 35 percent with some mitral valve leakage and left chamber enlargement. A new echo indicates an ejection fraction of 50 percent with minimal valve leakage, and only mild chamber enlargement. What could account for this improvement? PMID- 16721928 TI - When I'm stressed, I know my heart rate and blood pressure go up, and that's bad for my health. So how come it's considered healthy when my heart rate and blood pressure rise when I exercise? Logic suggests that a little stress now and then might provide the same benefits as exercise. PMID- 16721929 TI - Product recalls. Recalls: batteries for Disney personal DVD players... Trendset Originals girls' sweaters with drawstrings...and replacement program for magnetic building sets. PMID- 16721930 TI - Headache diary. PMID- 16721931 TI - Spotlight on Pierre Corvol, MD. PMID- 16721932 TI - A new cardiovascular research centre. PMID- 16721933 TI - A flap over bird flu.